PMID- 10787372 TI - Risk for obstructive sleep apnea. PMID- 10787373 TI - Risk for obstructive sleep apnea. PMID- 10787374 TI - Risk for obstructive sleep apnea. PMID- 10787375 TI - Nonfasting plasma total homocysteine level and mortality. PMID- 10787376 TI - Therapeutic plasma exchange. PMID- 10787377 TI - Potential new cardiovascular risk factors. PMID- 10787378 TI - Quotations on the wall. PMID- 10787379 TI - Serum cholesterol and low-density lipoprotein cholesterol levels as predictors of response to interferon therapy for chronic hepatitis C. PMID- 10787380 TI - Small dense low-density lipoprotein in Japanese men with coronary artery disease. PMID- 10787381 TI - Error in medicine: what have we learned? PMID- 10787382 TI - Occupational hygiene in developing countries: something to talk about? PMID- 10787383 TI - An evidence based EULAR meeting? PMID- 10787384 TI - Cutaneous leucocytoclastic vasculitis caused by cyclosporin A (Sandimmun) PMID- 10787385 TI - Alcohol and porphyrin metabolism. AB - Alcohol is a porphyrinogenic agent which may cause disturbances in porphyrin metabolism in healthy persons as well as biochemical and clinical manifestations of acute and chronic hepatic porphyrias. After excessive consumption of alcohol, a temporary, clinically asymptomatic secondary hepatic coproporphyrinuria is observable, which can become persistent in cases of alcohol-induced liver damage. Nowadays, the alcohol-liver-porphyrinuria syndrome is the first to be mentioned in secondary hepatic disturbances of porphyrin metabolism. Acute hepatic porphyrias (acute intermittent porphyria, variegate porphyria and hereditary coproporphyria) are considered to be molecular regulatory diseases, in contrast to non-acute, chronic hepatic porphyria, clinically appearing as porphyria cutanea tarda (PCT). Porphyrins do not accumulate in the liver in acute porphyrias, whereas in chronic hepatic porphyrias they do. Thus, chronic hepatic porphyria is a porphyrin-accumulation disease, whereas acute hepatic porphyrias are haem-pathway-dysregulation diseases, characterized in general by induction of delta-aminolevulinic acid synthase in the liver and excessive stimulation of the pathway without storage of porphyrins in the liver. The clinical expression of acute hepatic porphyrias can be triggered by alcohol, because alcohol augments the inducibility of delta-aminolevulinic acid synthase. In chronic hepatic porphyrias, however, which are already associated with liver damage, alcohol potentiates the disturbance of the decarboxylation of uro- and heptacarboxyporphyrinogen, which is followed by a hepatic accumulation of uro- and heptacarboxyporphyrin and their sometimes extreme urinary excretion. Especially in persons with a genetic deficiency of uroporphyrinogen decarboxylase, but also in patients with the so-called sporadic variety of PCT, alcohol is able to transform an asymptomatic coproporphyrinuria into PCT. Alcohol has many biochemical and clinical effects on porphyrin and haem synthesis both in humans and laboratory animals. Ethanol suppresses the activity of porphobilinogen synthase (synonym: delta-aminolevulinic acid dehydratase), uroporphyrinogen decarboxylase, coproporphyrinogen oxidase and ferrochelatase, whereas it induces the first and rate-limiting enzyme in the pathway, delta-aminolevulinic acid synthase and also porphobilinogen deaminase. Therefore, teetotalism is a therapeutically and prophylactically important measure in all types of hepatic porphyrias. PMID- 10787386 TI - Are anandamide and cannabinoid receptors involved in ethanol tolerance? A review of the evidence. AB - There have been significant developments towards the elucidation of molecular and cellular changes in neuronal second messenger pathways involved in the development of tolerance to and dependence on ethanol (EtOH). The long-term exposure to EtOH has been shown to affect several aspects of neuronal signal transduction as well as ligand-gated ion channels and receptor systems, including the receptors that are coupled to the superfamily of GTP binding regulatory proteins (G-proteins). The recent identification of a G-protein coupled receptor that was activated by delta-9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana, led to the discovery of endogenous agonists. One such agonist found to exist in mammalian brain was characterized to be an arachidonic acid (AA) metabolite and was named anandamide (AnNH). AnNH has been shown to bind specifically to the cannabinoid receptor (CB(1)) and mimic many of the pharmacological and behavioural effects of THC including tolerance development. The role of endocannabinoids and the CB(1) receptor signal transduction system in tolerance development to drugs of abuse has not been explored until recently. The findings presented in this review provide evidence for the first time that some of the pharmacological actions of EtOH including tolerance development may be mediated through participation of the endocannabinoid-CB(1) receptor signal transduction system. Recent studies have shown that chronic EtOH exposure produces downregulation of CB(1) receptors and an inhibition of CB(1) receptor agonist-stimulated GTPgammaS binding in mouse brain synaptic plasma membranes (SPM). The observed receptor downregulation results from the persistent stimulation of the receptors by the endogenous CB(1) receptor agonist AnNH, the synthesis of which is increased by chronic EtOH exposure. Further, the CB(1) receptor antagonist SR-141716A has been shown to block voluntary EtOH intake in rats and mice. Based on these studies, a hypothesis is presented to explain the possible involvement of the endocannabinoid system in the pharmacological and behavioural effects of EtOH. PMID- 10787387 TI - Acute ethanol treatment decreases intracellular calcium-ion transients in mouse single skeletal muscle fibres in vitro. AB - Alcohol misuse frequently leads to muscle weakness, which may also occur in the setting of acute and chronic alcoholic myopathies. At the cellular level, ethanol has been found to interfere with signalling mechanisms in cardiac myocytes, skeletal myotubes, and smooth muscle cells. In this study, we focused on the effects of ethanol on the intracellular calcium ([Ca(2+)](i)) transients responsible for excitation-contraction (EC) coupling in isolated mouse skeletal fibres loaded with the fluorescent Ca(2+) indicator fura-2. Following electrical stimulation, ethanol caused a significant reversible dose-dependent reduction in [Ca(2+)](i) transient amplitude, already significant at 100 mM ethanol (P = 0.03), without modifying resting [Ca(2+)](i). Evaluating the potential loci for the effects of ethanol, we indirectly measured sarcolemmal Ca(2+) entry by monitoring Mn(2+)-quenching of intracellular fura-2 via the nitrendipine sensitive Ca(2+) channels during electrical pacing. Ethanol at doses of 20 mM and greater caused a dose-dependent reduction in the rate of fura-2 quenching (all at P<0.05). Moreover, the intracellular pool of Ca(2+) releasable by caffeine was found to be reduced at a minimum of 300 mM ethanol (P = 0.05). We conclude that ethanol reduces the [Ca(2+)](i) transients underlying EC coupling in single mouse skeletal muscle fibres. This acute effect of ethanol was primarily due to an inhibitory effect of ethanol on sarcolemmal Ca(2+) influx via voltage-operated Ca(2+)-channels and, to a lesser extent, to a reduction in the Ca(2+) sarcoplasmic reticulum loading state. This inhibitory effect of ethanol may be implicated in the development of muscle weakness with alcohol consumption. PMID- 10787388 TI - Effect of adrenalectomy and exposure to corticosterone on alcohol intake in alcohol-preferring and alcohol-avoiding rat lines. AB - The daily fluid intake of male rats of the alcohol-preferring (AA) and alcohol avoiding (ANA) lines with simultaneous access to 10% (v/v) ethanol and water was determined during a baseline period (2 weeks), following adrenalectomy (1 week), and for 2 weeks following corticosterone treatment. The results showed that adrenalectomized AA rats decreased their ethanol intake compared to the sham operated AA controls and that treatment with corticosterone restored the intake of ethanol to that observed during the baseline period. In contrast to the AA rats, there were no alterations in ethanol intake after adrenalectomy and following corticosterone replacement in the ANA rats. These results suggest that corticosterone stimulates ethanol intake in animals with pronounced high preference for ethanol. PMID- 10787389 TI - The effect of ethanol on histone glycation in diabetic rats. AB - The glycation of liver histones was studied in rats with streptozotocin-induced diabetes, in ethanol (EtOH)-treated rats, and in EtOH-treated diabetic rats. In diabetes, the conditions of glucose-protein addition are more favourable extracellularly in serum and in erythrocytes than in the nucleus. This is indicated by the increased level of serum fructosamine and by the high level of glycated haemoglobin, while the glycation of intracellular histone is decreased. In the serum of diabetic rats, we found a relatively high acetaldehyde level, which resulted in elevated histone fluorescence. Fluorescence is an accepted marker of advanced glycation end-product (AGE), the intensity of which, according to our experiments is related not to the level of serum glucose, but to the level of acetaldehyde. The data obtained with histone proteins in diabetic rats treated with EtOH are in good agreement with the results of our earlier in vitro experimental results obtained with H1 histone: the reaction of the two aldehydes (glucose and acetaldehyde) in combination gives a lower glycohistone value than they do separately. PMID- 10787390 TI - Acute effects of growth hormone in alcohol-fed rats. AB - The present study examined whether administration in vivo of a maximally stimulating dose of growth hormone (GH) was capable of modulating selected aspects of the GH-insulin-like growth factor (IGF) system to the same extent in alcohol-fed and control animals. Rats were maintained on an alcohol-containing diet for 14 weeks, while control animals were fed isocalorically. After surgical implantation of a catheter in the carotid artery, rats were starved overnight. The next morning, rats were injected with recombinant human GH (500 microg/kg, s.c.) or an equal volume of saline at time 0 and 12 h. Blood samples were collected prior to GH and at 6, 12 and 24 h thereafter; tissues were collected at the end of the study. Time-matched control and alcohol-fed rats not receiving GH were also included. Although the plasma concentrations of both total and free IGF I were decreased 30-40% in alcohol-fed rats, the ability of GH to elevate circulating IGF-I was not diminished. GH was equally effective at increasing IGF I peptide levels in both liver and skeletal muscle. GH also produced comparable increases in IGF-I mRNA in muscle in both groups. Hepatic GH receptor (GHR) peptide levels were not significantly altered by either alcohol or GH. Alcohol feeding decreased plasma levels of IGF binding protein (IGFBP)-3 and increased IGFBP-1, and GH did not significantly alter this profile. Hepatic expression of suppressor of cytokine signalling (SOCS-3) mRNA was not different between the groups. However, SOCS-3 mRNA was increased by approximately 50% in control animals in response to GH, but remained unchanged in alcohol-fed rats. These data indicate that the decrease in hepatic IGF-I synthesis and plasma IGF-I observed in alcohol-fed rats was independent of a change in GHR levels. In contrast, the ability of a maximally stimulating dose of GH to modulate selected biological responses in vivo was not impaired by chronic alcohol consumption and was associated with a lack of a GH-induced increase in SOCS-3 mRNA. PMID- 10787391 TI - Regulation of manganese superoxide dismutase (MnSOD) in chronic experimental alcoholism: effects of vitamin E-supplemented and -deficient diets. AB - In order to investigate the pathogenic mechanism responsible for liver injury associated with chronic alcoholism, we studied the effects of different dietary vitamin E levels in chronically ethanol (EtOH)-fed rats on the activity and mRNA regulation of the manganese superoxide dismutase (MnSOD) enzyme. Evidence is accumulating that intermediates of oxygen reduction may in fact be associated with the development of alcoholic liver disease. Since low vitamin E liver content seems to potentiate EtOH-linked oxidative stress, we studied the effect of EtOH treatment in livers from rats fed a diet deficient or supplemented with vitamin E. Chronic EtOH feeding enhanced hepatic consumption of vitamin E in both groups of EtOH-treated animals, irrespectively of the vitamin E level of the basal diet and the effect was observed in both the microsomal and mitochondrial fractions. Both EtOH-fed groups exhibited increased MnSOD gene expression, while the enzyme activity was enhanced only in the vitamin E-deprived group of EtOH treated animals. The significant increase in manganese liver content found only in this last group could explain the rise of enzyme activity. In fact, in the absence of a parallel increase of the prosthetic ion manganese, MnSOD mRNA induction was not accompanied by a higher enzymatic activity. These findings support the role of oxidative alteration in the EtOH-induced chronic hepatotoxicity in which MnSOD response might represent a primary defence mechanism against the damaging effect of oxygen radical species. PMID- 10787392 TI - Comparison of the formation of proteins modified by direct and indirect ethanol metabolites in the liver and blood of rats fed the Lieber-DeCarli liquid diet. AB - It has been proposed that proteins modified by ethanol metabolites, such as acetaldehyde (AcH) or alpha-hydroxyethyl radicals (HER) may be an important step in the aetiology of alcoholic liver disease. Furthermore, it has also been suggested that these modified proteins may act as a marker of ethanol intake. In this study, we have measured the generation of various types of modified proteins in the liver and blood of ethanol-fed rats. Multiple types of protein modification were observed in the livers of the ethanol-fed rats. In each case, the level of modification increased over the first 6 weeks of ethanol feeding, but reached a plateau by 10 weeks. In contrast to the liver, elevated levels of proteins modified by malondialdehyde were not seen in the plasma of ethanol-fed animals, whereas elevated levels of modification due to AcH and HER were observed. In haemolysates from these animals, only modification due to AcH was seen. Further investigation of the modification of plasma proteins showed that albumin, a protein produced in the liver, carried all the types of modification investigated, whereas immunoglobulin G, a protein derived from an extra-hepatic source, only carried modifications due to acetaldehyde. This study demonstrates for the first time that modification of plasma proteins by ethanol metabolites can occur at both intra- and extra-hepatic sites. PMID- 10787393 TI - Clinical correlates of cigarette smoking and nicotine dependence in alcohol dependent men and women. The Collaborative Study Group on the Genetics of Alcoholism. AB - This paper examines the clinical characteristics associated with tobacco use and nicotine dependence in a large sample of alcohol-dependent subjects. The goal was to determine if the characteristics of the alcohol use history were associated with the smoking status, even after controlling for additional characteristics, such as the antisocial personality disorder, other drug dependence and gender. As part of the Collaborative Study on the Genetics of Alcoholism, a semi-structured interview, including a detailed history of alcohol and tobacco use, was administered to 1005 alcohol-dependent men and women, made up of 658 (65.5%) current smokers, 167 (16.6%) former smokers, and 180 (17.9%) non-smokers. Among former smokers, 50.3%, and among current smokers, 72.8% had ever been nicotine dependent (DSM-III-R). Current smokers and nicotine-dependent subjects had a greater severity of alcohol dependence, even as evaluated through logistic regression analyses in which gender and associated diagnoses were considered. The data also enabled us to study the relationships among depression, nicotine dependence, and alcohol dependence, with most of the correlation occurring for substance-induced, not independent, mood disorders. PMID- 10787394 TI - United Kingdom Multicentre Acamprosate Study (UKMAS): a 6-month prospective study of acamprosate versus placebo in preventing relapse after withdrawal from alcohol. AB - A 6-month randomized controlled study of acamprosate versus placebo in preventing relapse following withdrawal from alcohol was undertaken in 20 centres throughout the UK. Patients diagnosed as alcohol-dependent and detoxified within the preceding 5 weeks were randomly assigned to treatment with either acamprosate (A) 666 mg three times/day or identical placebo (P). A total of 664 patients were screened; 581 were entered into the treatment phase. One-third were episodic drinkers, 84% were male, 44% were unmarried and 48% were unemployed. Medication was first taken on average 24 days after the start of detoxification; 32% of patients had already relapsed by this time. The 6-month study period was completed by 35% of patients; adverse events led to withdrawal of a further 14% (A) and 9% (P) respectively. Compliance was poor in that, by the end of the second week, only 57% of patients were judged to be taking at least 90% of their tablets. The mean total of abstinent days achieved was 77 (A) and 81 (P). Complete abstinence for 6 months was achieved by 12% (A) and 11% (P); drinking remained within controlled limits in a further 3% (A) and 6% (P). An effect of acamprosate on consumption was not seen when subgroups, including those defined by the Lesch typology, were analysed separately. However, the mean percentage reduction in craving for alcohol measured on a visual analogue scale was greater in the acamprosate, than placebo, patients at week 2 and week 4 (P<0.001) and the mean decrease in the Hamilton Anxiety score at the 4th week was greater in the acamprosate than placebo patients (P = 0.017). In comparison with other published trials of acamprosate, patients started study medication after a longer time following detoxification, had more often recommenced drinking before medication was started and had a higher drop-out rate, and this might have contributed to the lack of a treatment effect in this study. PMID- 10787395 TI - Alcohol withdrawal and hypokalaemia: a case report. AB - A case is presented where a 25-year-old man developed a serious hypokalaemia (K(+) 2.2 mmol/l) during alcohol withdrawal, despite intravenous saline treatment and normal feeding. As hypokalaemia can be symptom-free, we want to draw attention to the combination of vomiting, malnutrition and alcohol withdrawal, as these can cause lethal complications. We therefore recommend that potassium serum level should be routinely monitored during alcohol withdrawal, even when this is being managed in the community. PMID- 10787396 TI - Risk factors for alcohol dependence: a case-control study. AB - Several possible risk factors for ICD-10 alcohol dependence were studied by comparing cases (117 men, 188 women) with controls (248 men, 300 women). Logistic regression analyses showed that parental alcohol problems and high trait anxiety were significantly related to high occurrence of alcohol dependence in both men and women. In women, high antisocial behaviour, high impulsivity, and high externality were also related to high occurrence of alcohol dependence. High facial flushing and high stimulation when intoxicated were related to low occurrence of alcohol dependence in both men and women. In men, this was also the case for high social support. Several interactions were observed. In contrast to earlier studies, there was no significant association between alcohol dependence and left-handedness. PMID- 10787397 TI - Alcohol impairs speed of information processing and simple and choice reaction time and differentially impairs higher-order cognitive abilities. AB - Previous research has demonstrated that alcohol impairs information processing. However, it is unknown whether this impairment is on all stages of information processing, or on the early, rather than on the later, stages of information processing. Thus, the aim of the present study was to examine the effects of orally administered alcohol on both the early and the later stages of information processing. The present study assessed inspection time (IT), simple reaction time, choice reaction time and cognitive ability (Wechsler Adult Intelligence Scale - Revised) in 16 adult participants in both alcohol and placebo conditions. IT (a measure of the early stages of information processing) was used as a predictor variable in a linear regression to examine whether a disruption of the early stages of information processing accounted for changes in total information processing after alcohol administration. Results indicated that alcohol significantly slowed total information processing, independently of the early stages of information processing. PMID- 10787398 TI - Acamprosate and relapse prevention in the treatment of alcohol dependence: a placebo-controlled study. AB - The objective of this study was to compare acamprosate with placebo in the treatment of alcohol-dependent patients during a 6-month post-detoxification treatment and a 3-month medication-free follow-up. Patients (n = 330) were detoxified and randomized to treatment with acamprosate (1998 mg/day) or placebo within an out-patient programme including medical counselling, psychotherapy and self-help groups. The main outcome criterion was drinking behaviour as assessed by: abstinence/relapse ratio, cumulative abstinence duration (CAD) and the period of continued abstinence. Anxiety, depression and craving were also monitored. Intention to treat (ITT) statistical principles were followed. Twenty-five per cent of patients dropped out over the first 6 months. At the end of the treatment period, the abstinence rate was 57.9% for acamprosate and 45.2% for placebo (P = 0.03). The CAD was 110+/-77 days for acamprosate and 89+/-77 days for placebo (P = 0.016). Patients on acamprosate had a higher continuous abstinence rate and experienced less severe relapses. No differential effect was noted for anxiety, depression or craving. Treatment remained positive, but not significant, 3 months after termination of study medication. No significant difference in adverse events was noted between treatment groups. Acamprosate treatment over 180 days was consistently more effective than placebo to maintain abstinence and to diminish relapse severity. PMID- 10787399 TI - Alcohol-dependent patients with neuroendocrine evidence for reduced dopamine d(2) receptor function have decreased platelet monoamine oxidase-B activity. PMID- 10787400 TI - Is daily single dosage of diazepam as effective as chlordiazepoxide in divided doses in alcohol withdrawal--a pilot study. PMID- 10787401 TI - Reversal of ethanol-induced hepatic steatosis and lipid peroxidation by taurine: a study in rats. PMID- 10787402 TI - The cytokine portion of p43 occupies a central position within the eukaryotic multisynthetase complex. AB - Multicellular eukaryotes contain a macromolecular assembly of nine aminoacyl-tRNA synthetase activities and three auxiliary proteins. One of these, p43, is the precursor of endothelial monocyte-activating polypeptide II (EMAP II), an inflammatory cytokine involved in apoptotic processes. As a step toward understanding this paradoxical association, the EMAP II portion of p43 has been localized within the rabbit reticulocyte multisynthetase complex. Immunoblot analysis demonstrates strong reaction of anti-EMAP II antiserum with p43, as well as cross-reactivity with isoleucyl-tRNA synthetase. Electron microscopic images of immunocomplexes show two antibody binding sites. The primary site is near the midpoint of the multisynthetase complex at the intersection of the arms with the base. This site near the lower edge of the central cleft is assigned to the C terminal cytokine portion of p43. The secondary site of antibody binding is in the base of the particle and maps the location of isoleucyl-tRNA synthetase. These data allow refinement of the three-domain model of polypeptide distribution within the multisynthetase complex. Moreover, the central location of p43/EMAP II suggests a role for this polypeptide in optimizing normal function and in rapid disruption of essential cellular machinery when apoptosis is signaled. PMID- 10787403 TI - The spider toxin omega-Aga IIIA defines a high affinity site on neuronal high voltage-activated calcium channels. AB - The spider toxin omega-agatoxin IIIA (omega-Aga-IIIA) is a potent inhibitor of high voltage-activated calcium currents in the mammalian brain. To establish the biochemical parameters governing its action, we radiolabeled the toxin and examined its binding to native and recombinant calcium channels. In experiments with purified rat synaptosomal membranes, both kinetic and equilibrium data demonstrate one-to-one binding of omega-Aga-IIIA to a single population of high affinity sites, with K(d) = approximately 9 pm and B(max) = approximately 1.4 pmol/mg protein. Partial inhibition of omega-Aga-IIIA binding by omega-conotoxins GVIA, MVIIA, and MVIIC identifies N and P/Q channels as components of this population. omega-Aga-IIIA binds to recombinant alpha(1B) and alpha(1E) calcium channels with a similar high affinity (K(d) = approximately 5-9 pm) in apparent one-to-one fashion. Results from recombinant alpha(1B) binding experiments demonstrate virtually identical B(max) values for omega-Aga-IIIA and omega conotoxin MVIIA, providing further evidence for a one-to-one stoichiometry of agatoxin binding to calcium channels. The combined evidence suggests that omega Aga-IIIA defines a unique, high affinity binding site on N-, P/Q-, and R-type calcium channels. PMID- 10787404 TI - Multiple zinc binding sites in retinal rod cGMP phosphodiesterase, PDE6alpha beta. AB - The photoreceptor cGMP phosphodiesterase (PDE6) plays a key role in vertebrate vision, but its enzymatic mechanism and the roles of metal ion co-factors have yet to be determined. We have determined the amount of endogenous Zn(2+) in rod PDE6 and established a requirement for tightly bound Zn(2+) in catalysis. Purified PDE6 contained 3-4-g atoms of zinc/mole, consistent with an initial content of two tightly bound Zn(2+)/catalytic subunit. PDE with only tightly bound Zn(2+) and no free metal ions was inactive, but activity was fully restored by Mg(2+), Mn(2+), Co(2+), or Zn(2+). Mn(2+), Co(2+), and Zn(2+) also induced aggregation and inactivation at higher concentrations and longer times. Removal of 93% of the tightly bound Zn(2+) by treatment with dipicolinic acid and EDTA at pH 6.0 resulted in almost complete loss of activity in the presence of Mg(2+). This activity loss was blocked almost completely by Zn(2+), less potently by Co(2+) and almost not at all by Mg(2+), Mn(2+), or Cu(2+). The lost activity was restored by the addition of Zn(2+), but Co(2+) restored only 13% as much activity, and other metals even less. Thus tightly bound Zn(2+) is required for catalysis but could also play a role in stabilizing the structure of PDE6, whereas distinct sites where Zn(2+) is rapidly exchanged are likely occupied by Mg(2+) under physiological conditions. PMID- 10787405 TI - Protein-tyrosine phosphatase reduces the number of apical small conductance K+ channels in the rat cortical collecting duct. AB - Previous studies have demonstrated that an increase in the activity of protein tyrosine kinase (PTK) is involved in the down-regulation of the activity of apical small conductance K(+) (SK) channels in the cortical collecting duct (CCD) from rats on a K(+)-deficient diet (). We used the patch clamp technique to investigate the role of protein-tyrosine phosphatase (PTP) in the regulation of the activity of SK channels in the CCD from rats on a high K(+) diet. Western blot analysis indicated that PTP-1D is expressed in the renal cortex. Application of 1 microm phenylarsine oxide (PAO) or 1 mm benzylphosphonic acid, agents that inhibit PTP, reversibly reduced channel activity by 95%. Pretreatment of CCDs with PAO for 30 min decreased the mean NP(o) reversibly from control value 3.20 to 0.40. Addition of 1 microm herbimycin A, an inhibitor of PTK, had no significant effect on channel activity in the CCDs from rats on a high K(+) diet. However, herbimycin A abolished the inhibitory effect of PAO, indicating that the effect of PAO is the result of interaction between PTK and PTP. Addition of brefeldin A, an agent that blocks protein trafficking from Golgi complex to the membrane, had no effect on channel activity. Moreover, application of colchicine, a microtubule inhibitor, or paclitaxel, a microtubule stabilizer, had no effect on channel activity. In contrast, PAO still reduced channel activity in the presence of brefeldin A, colchicine, or paclitaxel. Furthermore, the effect of PAO on channel activity was absent when the tubules were bathed in 16% sucrose containing bath solution or treated with concanavalin A. We conclude that PTP is involved in the regulation of the activity of SK channels and that inhibition of PTP may facilitate the internalization of the SK channels. PMID- 10787406 TI - The Saccharomyces cerevisiae RuvB-like protein, Tih2p, is required for cell cycle progression and RNA polymerase II-directed transcription. AB - Two highly conserved RuvB-like putative DNA helicases, p47/TIP49b and p50/TIP49a, have been identified in the eukaryotes. Here, we study the function of Saccharomyces cerevisiae TIH2, which corresponds to mammalian p47/TIP49b. Tih2p is required for vegetative cell growth and localizes in the nucleus. Immunoprecipitation analysis revealed that Tih2p tightly interacts with Tih1p, the counterpart of mammalian p50/TIP49a, which has been shown to interact with the TATA-binding protein and the RNA polymerase II holoenzyme complex. Furthermore, the mutational study of the Walker A motif, which is required for nucleotide binding and hydrolysis, showed that this motif plays indispensable roles in the function of Tih2p. When a temperature-sensitive tih2 mutant, tih2 160, was incubated at the nonpermissive temperature, cells were rapidly arrested in the G(1) phase. Northern blot analysis revealed that Tih2p is required for transcription of G(1) cyclin and of several ribosomal protein genes. The similarities between the mutant phenotypes of tih2-160 and those of taf145 mutants suggest a role for TIH2 in the regulation of RNA polymerase II-directed transcription. PMID- 10787407 TI - Regulation of F-actin and endoplasmic reticulum organization by the trimeric G protein Gi2 in rat hepatocytes. Implication for the activation of store-operated Ca2+ inflow. AB - The roles of the heterotrimeric G-protein, G(i2), in regulating the actin cytoskeleton and the activation of store-operated Ca(2+) channels in rat hepatocytes were investigated. Galpha(i2) was principally associated with the plasma membrane and microsomes. Both F-actin and Galpha(i2) were detected by Western blot analysis in a purified plasma membrane preparation, the supernatant and pellet obtained by treating the plasma membrane with Triton X-100, and after depolymerization and repolymerization of F-actin in the Triton X-100-insoluble pellet. Actin in the Triton X-100-soluble supernatant co-precipitated with Galpha(i2) using either anti-Galpha(i2) or anti-actin antibodies. The principally cortical location of F-actin in hepatocytes cultured for 0.5 h changed to a pericanalicular distribution over a further 3.5 h. Some Galpha(i2) co-localized with F-actin at the plasma membrane. Pretreatment with pertussis toxin ADP ribosylated 70-80% of Galpha(i2) in the plasma membrane and microsomes, prevented the redistribution of F-actin, caused redistribution and fragmentation of the endoplasmic reticulum, and inhibited vasopressin-stimulated Ca(2+) inflow. It is concluded that (i) a significant portion of hepatocyte Galpha(i2) associates with, and regulates the arrangement of, cortical F-actin and the endoplasmic reticulum and (ii) either or both of these regulatory roles are likely to be required for normal vasopressin activation of Ca(2+) inflow. PMID- 10787408 TI - Identification of p130cas as an in vivo substrate of receptor protein-tyrosine phosphatase alpha. AB - We have employed a substrate trapping strategy to identify physiological substrates of the receptor protein-tyrosine phosphatase alpha (RPTPalpha). Here we report that a substrate-trapping mutant of the RPTPalpha membrane proximal catalytic domain (D1), RPTPalpha-D1-C433S, specifically bound to tyrosine phosphorylated proteins from pervanadate-treated cells. The membrane distal catalytic domain of RPTPalpha (D2) and mutants thereof did not bind to tyrosine phosphorylated proteins. The pattern of tyrosine-phosphorylated proteins that bound to RPTPalpha-D1-C433S varied between cell lines, but a protein of approximately 130 kDa was pulled down from every cell line. This protein was identified as p130(cas). Tyrosine-phosphorylated p130(cas) from fibronectin stimulated NIH3T3 cells bound to RPTPalpha-D1-C433S as well, suggesting that p130(cas) is a physiological substrate of RPTPalpha. RPTPalpha dephosphorylated p130(cas) in vitro, and RPTPalpha co-localized with a subpopulation of p130(cas) to the plasma membrane. Co-transfection experiments with activated SrcY529F, p130(cas), and RPTPalpha or inactive, mutant RPTPalpha indicated that RPTPalpha dephosphorylated p130(cas) in vivo. Tyrosine-phosphorylated epidermal growth factor receptor was not dephosphorylated by RPTPalpha under these conditions, suggesting that p130(cas) is a specific substrate of RPTPalpha in living cells. In conclusion, our results provide evidence that p130(cas) is a physiological substrate of RPTPalpha in vivo. PMID- 10787409 TI - Characterization of 1-deoxy-D-xylulose 5-phosphate reductoisomerase, an enzyme involved in isopentenyl diphosphate biosynthesis, and identification of its catalytic amino acid residues. AB - 1-Deoxy-d-xylulose 5-phosphate (DXP) reductoisomerase, which simultaneously catalyzes the intramolecular rearrangement and reduction of DXP to form 2-C methyl-d-erythritol 4-phosphate, constitutes a key enzyme of an alternative mevalonate-independent pathway for isopentenyl diphosphate biosynthesis. The dxr gene encoding this enzyme from Escherichia coli was overexpressed as a histidine tagged protein and characterized in detail. DNA sequencing analysis of the dxr genes from 10 E. coli dxr-deficient mutants revealed base substitution mutations at four points: two nonsense mutations and two amino acid substitutions (Gly(14) to Asp(14) and Glu(231) to Lys(231)). Diethyl pyrocarbonate treatment inactivated DXP reductoisomerase, and subsequent hydroxylamine treatment restored the activity of the diethyl pyrocarbonate-treated enzyme. To characterize these defects, we overexpressed the mutant enzymes G14D, E231K, H153Q, H209Q, and H257Q. All of these mutant enzymes except for G14D were obtained as soluble proteins. Although the purified enzyme E231K had wild-type K(m) values for DXP and NADPH, the mutant enzyme had less than a 0.24% wild-type k(cat) value. K(m) values of H153Q, H209Q, and H257Q for DXP increased to 3.5-, 7.6-, and 19-fold the wild-type value, respectively. These results indicate that Glu(231) of E. coli DXP reductoisomerase plays an important role(s) in the conversion of DXP to 2-C-methyl-d-erythritol 4-phosphate, and that His(153), His(209), and His(257), in part, associate with DXP binding in the enzyme molecule. PMID- 10787410 TI - A chimeric transmembrane domain directs endothelial nitric-oxide synthase palmitoylation and targeting to plasmalemmal caveolae. AB - The endothelial nitric-oxide synthase (eNOS), a key signaling protein, undergoes a series of covalent modifications, including co-translational N-myristoylation at Gly(2), as well as post-translational thiopalmitoylation at Cys(15) and Cys(26). Myristoylation of eNOS is required for the subsequent palmitoylation of the enzyme, and both acylations are required for the efficient subcellular targeting of eNOS to plasmalemmal caveolae. We constructed chimeric cDNAs encoding proteins comprised of various acylation-deficient eNOS mutants fused at their N termini to the hydrophobic transmembrane domain of the glycoprotein CD8 and characterized these constructs in transient transfection experiments in COS-7 cells. One construct (termed CD8-myr(-)eNOS) encodes a fusion protein comprised of the eNOS myristoylation-deficient mutant coupled to the CD8 transmembrane domain. In biosynthetic labeling experiments using [(3)H]palmitic acid, we found that the CD8-myr(-)eNOS chimera undergoes palmitoylation. Subcellular fractionation showed that the CD8-myr(-)eNOS chimera is targeted to caveolae. We also constructed and characterized a cDNA encoding the CD8 transmembrane domain fused to the palmitoylation-deficient mutant eNOS (in which Cys(15) and Cys(26) are changed to serine). This chimera (termed CD8-myr(-).palm(-)eNOS) did not undergo palmitoylation, indicating that the palmitoylation seen with the CD8. myr(-)eNOS fusion protein occurs on the same residues as in the wild-type enzyme. Importantly, the CD8-myr(-).palm(-)eNOS fusion protein remained efficiently targeted to caveolae, in contrast to the palm(-)eNOS mutant lacking the CD8 transmembrane domain, which has nominal caveolar localization. A construct encoding the CD8 transmembrane domain alone was insufficient for selective targeting to caveolae. These results indicate that membrane targeting per se, but not necessarily myristoylation, is sufficient for eNOS palmitoylation and localization to plasmalemmal caveolae, and suggest further that sequences within eNOS itself, in addition to its palmitoylation sites, facilitate the selective localization of the enzyme within caveolae. PMID- 10787411 TI - Specific recognition of androgens by their nuclear receptor. A structure-function study. AB - Androgens, like progestins, are 3-ketosteroids with structural differences restricted to the 17beta substituent in the steroid D-ring. To better understand the specific recognition of ligands by the human androgen receptor (hAR), a homology model of the ligand-binding domain (LBD) was constructed based on the progesterone receptor LBD crystal structure. Several mutants of residues potentially involved in the specific recognition of ligands in the hAR were constructed and tested for their ability to bind agonists. Their transactivation capacity in response to agonist (R1881) and antagonists (cyproterone acetate, hydroxyflutamide, and ICI 176344) was also measured. Substitution of His(874) by alanine, only marginally impairs the ligand-binding and transactivation capacity of the hAR receptor. In contrast, mutations of Thr(877) and, to a greater extent, Asn(705) perturb ligand recognition, alter transactivation efficiency, and broaden receptor specificity. Interestingly, the N705A mutant acquires progesterone receptor (PR) properties for agonist ligands but, unlike wild type AR and PR, loses the capacity to repress transactivation with nonsteroidal antagonists. Models of the hAR.LBD complexes with several ligands are presented, which suggests new directions for drug design. PMID- 10787412 TI - Aspartate 351 of estrogen receptor alpha is not crucial for the antagonist activity of antiestrogens. AB - The antagonist activity of antiestrogens is due to the presence of a long carbon side chain at positions 7alpha or 11beta or equivalent on their steroid or steroid-like skeletons. These side chains establish hydrophobic interactions with amino acids of the estrogen receptor alpha (ERalpha) ligand binding domain. In addition, a hydrogen bond formed between amino acid Asp-351 and the tertiary amine present at the end of the side chain of partial antiestrogens is considered to be crucial for their antiestrogenicity. Here, we have investigated the role of Asp-351 in antiestrogen action in transiently transfected HeLa and MDA-MB-231 cells. Our results indicate that disruption of the negative charge at position 351 does not increase the agonist activity of partial antiestrogens and thus that the hydrogen bond with the antiestrogen side chain is not determinant in positioning the side chain in an antagonist position. The negative charge at position 351 was not required for transcriptional activity in the presence of hormone, but its presence was necessary for basal activity of the wild-type receptor and constitutive activities of mutants L536P and Y537A, suggesting a role of Asp-351 in stabilizing the active conformation of ERalpha. This stabilizing role of Asp-351 could be due to interaction of Asp-351 with the amide group of the peptide bond between Leu-539 and Leu-540 in helix 12 observed in the active conformation of the ERalpha ligand binding domain. PMID- 10787413 TI - Regulation of high affinity nickel uptake in bacteria. Ni2+-Dependent interaction of NikR with wild-type and mutant operator sites. AB - Escherichia coli actively imports nickel via the ATP-dependent NikABCDE permease. NikR, a protein of the ribbon-helix-helix family of transcription factors, represses expression of the nikABCDE operon in the presence of excessive concentrations of intracellular nickel. Here, the NikR operator site is identified within the nikABCDE promoter by footprinting and mutational analyses. The operator consists of two dyad-symmetric 5'-GTATGA-3' recognition sequences separated by 16 base pairs. Mutations in the GTATGA sequences reduce NikR binding affinity in vitro and reduce repression of a P(nik)-lacZ fusion in vivo. Moreover, NikR is shown to be a direct sensor of nickel ions. Strong operator binding requires the continual presence of 20-50 micrometer nickel, indicating the presence of a low affinity nickel-binding site, and NikR dimers also contain two high affinity nickel-binding sites. In addition to both GTATGA sites and nickel, high affinity operator binding also requires the C-terminal domain of NikR. PMID- 10787414 TI - Expression and differential polarization of the reduced-folate transporter-1 and the folate receptor alpha in mammalian retinal pigment epithelium. AB - The differential polarized distribution of the reduced- folate transporter (RFT 1) and folate receptor alpha (FRalpha), the two proteins involved in the transport of folate, has been characterized in normal mouse retinal pigment epithelium (RPE) and in cultured human RPE cells. RPE cells mediate the vectorial transfer of nutrients from choroidal blood to neural retina. Whereas FRalpha is known to be present in many cell types of the neural retina, in situ hybridization analysis in the present study demonstrated that RFT-1 is present only in RPE. Laser-scanning confocal microscopy using antibodies specific for RFT 1 demonstrated an apical distribution of this protein in cultured human and intact mouse RPE, which contrasts with the basolateral distribution of FRalpha in these cells. The expression of RFT-1 in the RPE cell apical membrane was confirmed by functional studies with purified apical membrane vesicles from bovine RPE. These studies, done with N(5)-methyltetrahydrofolate (the predominant folate derivative in blood) and folate as substrates, have shown that RFT-1 functions in a Na(+)- and C1(-)-independent manner. The transporter is specific for folate and its analogs. A transmembrane H(+) gradient influences the transport function of this protein markedly; the transport mechanism is likely to be either folate/H(+) co-transport or folate/OH(-) exchange. Based on the differential polarization of FRalpha and RFT-1 in RPE, we suggest that these two proteins work in a concerted manner to bring about the vectorial transfer of folate across the RPE cell layer from the choroidal blood to the neural retina. This constitutes the first report of the differential polarization of the two folate transport proteins in any polarized epithelium. PMID- 10787416 TI - Molecular cloning and functional expression of KCNQ5, a potassium channel subunit that may contribute to neuronal M-current diversity. AB - We have isolated KCNQ5, a novel human member of the KCNQ potassium channel gene family that is differentially expressed in subregions of the brain and in skeletal muscle. When expressed in Xenopus oocytes, KCNQ5 generated voltage dependent, slowly activating K(+)-selective currents that displayed a marked inward rectification at positive membrane voltages. KCNQ5 currents were insensitive to the K(+) channel blocker tetraethylammonium but were strongly inhibited by the selective M-current blocker linopirdine. Upon coexpression with the structurally related KCNQ3 channel subunit, current amplitudes increased 4-5 fold. Compared with homomeric KCNQ5 currents, KCNQ3/KCNQ5 currents also displayed slower activation kinetics and less inward rectification, indicating that KCNQ5 combined with KCNQ3 to form functional heteromeric channel proteins. This functional interaction between KCNQ5 and KCNQ3, a component of the M-channel, suggests that KCNQ5 may contribute to a diversity of heteromeric channels underlying native neuronal M-currents. PMID- 10787417 TI - Vascular endothelial growth factor up-regulates ICAM-1 expression via the phosphatidylinositol 3 OH-kinase/AKT/Nitric oxide pathway and modulates migration of brain microvascular endothelial cells. AB - Endothelium of the cerebral blood microvessels, which constitutes the major component of the blood-brain barrier, controls leukocyte and metastatic cancer cell adhesion and trafficking into the brain parenchyma. In this study, using rat primary brain microvascular endothelial cells (BMEC), we demonstrate that the vascular endothelial growth factor (VEGF), a potent promoter of angiogenesis, up regulates the expression of the intracellular adhesion molecule-1 (ICAM-1) through a novel pathway that includes phosphatidylinositol 3 OH-kinase (PI3K), AKT, and nitric oxide (NO), resulting in the migration of BMEC. Upon VEGF treatment, AKT is phosphorylated in a PI3K-dependent manner. AKT activation leads to NO production and release and activation-deficient AKT attenuates NO production stimulated by VEGF. Transfection of the constitutive myr-AKT construct significantly increased basal NO release in BMEC. In these cells, VEGF and the endothelium-derived NO synergistically up-regulated the expression of ICAM-1, which was mediated by the PI3K pathway. This activity was blocked by the PI3K specific inhibitor, wortmannin. Furthermore, VEGF and NO significantly increased BMEC migration, which was mediated by the up-regulation of ICAM-1 expression and was dependent on the integrity of the PI3K/AKT/NO pathway. This effect was abolished by wortmannin, by the specific ICAM-1 antibody, by the specific inhibitor of NO synthase, N(G)-l-monomethyl-arginine (l-NMMA) or by a combination of wortmannin, ICAM-1 antibody, and l-NMMA. These findings demonstrate that the angiogenic factor VEGF up-regulates ICAM-1 expression and signals to ICAM-1 as an effector molecule through the PI3K/AKT/NO pathway, which leads to brain microvessel endothelial cell migration. These observations may contribute to a better understanding of BMEC angiogenesis and the physiological as well as pathophysiological function of the blood-brain barrier, whose integrity is crucial for normal brain function. PMID- 10787418 TI - Heat-induced relocalization of protein kinase CK2. Implication of CK2 in the context of cellular stress. AB - Among various other roles described so far, protein kinase CK2 has been involved in cell cycle, proliferation, and development. Here, we show that in response to specific stresses (heat shock or UV irradiation), a pool of the cellular CK2 content relocalizes in a particular nuclear fraction, increasing the activity of the kinase there. Electron microscopic analysis shows that upon heat shock, CK2alpha and CK2beta subunits are both detected in similar speckle structures occurring in the interchromatin space but are differentially targeted inside the nucleolus. This CK2 relocalization process takes place in a time- and dose dependent manner and is reversible upon recovery at 37 degrees C. Altogether, this work suggests CK2 be involved in the response to physiological stress in higher eukaryotic cells. PMID- 10787419 TI - Telomere shortening is proportional to the size of the G-rich telomeric 3' overhang. AB - Most normal diploid human cells do not express telomerase activity and are unable to maintain telomere length with ongoing cell divisions. We show that the length of the single-stranded G-rich telomeric 3'-overhang is proportional to the rate of shortening in four human cell types that exhibit different rates of telomere shortening in culture. These results provide direct evidence that the size of the G-rich overhang is not fixed but subject to regulation. The potential ability to manipulate this rate has profound implications both for slowing the rate of replicative aging in normal cells and for accelerating the rate of telomere loss in cancer cells in combination with anti-telomerase therapies. PMID- 10787420 TI - Crystal structure of the allergen Equ c 1. A dimeric lipocalin with restricted IgE-reactive epitopes. AB - The three-dimensional structure of the major horse allergen Equ c 1 has been determined at 2.3 A resolution by x-ray crystallography. Equ c 1 displays the typical fold of lipocalins, a beta-barrel flanked by a C-terminal alpha-helix. The space between the two beta-sheets of the barrel defines an internal cavity that could serve, as in other lipocalins, for the binding and transport of small hydrophobic ligands. Equ c 1 crystallizes in a novel dimeric form, which is distinct from that observed in other lipocalin dimers and corresponds to the functional form of the allergen. Binding studies of point mutants of the allergen with specific monoclonal antibodies raised in mouse and IgE serum from horse allergic patients allowed to identify putative B cell antigenic determinants. In addition, total inhibition of IgE serum recognition by a single specific monoclonal antibody revealed the restricted nature of the IgE binding target on the molecular surface of Equ c 1. PMID- 10787421 TI - Transport of proteins into cryptomonads complex plastids. AB - Complex plastids, found in many alga groups, are surrounded by three or four membranes. Therefore, proteins of the complex plastids, which are encoded in the cell nucleus, must cross three or four membranes during transport to the plastid. To study this process we have developed a method for isolating transport competent two membrane-bound plastids derived from the complex plastids of the cryptophyte Guillardia theta. This in vitro protein import system provides the first non-heterologous system for studying the import of proteins into four membrane complex plastids. We use our import system as well as canine microsomes to demonstrate in the case of cryptomonads how nuclear proteins pass the first nucleomorph-encoded proteins the third and fourth membrane and discuss the potential mechanisms for protein transport across the second membrane. PMID- 10787422 TI - Regulation of peroxisome size and number by fatty acid beta -oxidation in the yeast yarrowia lipolytica. AB - The Yarrowia lipolytica MFE2 gene encodes peroxisomal beta-oxidation multifunctional enzyme type 2 (MFE2). MFE2 is peroxisomal in a wild-type strain but is cytosolic in a strain lacking the peroxisomal targeting signal-1 (PTS1) receptor. MFE2 has a PTS1, Ala-Lys-Leu, that is essential for targeting to peroxisomes. MFE2 lacking a PTS1 can apparently oligomerize with full-length MFE2 to enable targetting to peroxisomes. Peroxisomes of an oleic acid-induced MFE2 deletion strain, mfe2-KO, are larger and more abundant than those of the wild type strain. Under growth conditions not requiring peroxisomes, peroxisomes of mfe2-KO are larger but less abundant than those of the wild-type strain, suggesting a role for MFE2 in the regulation of peroxisome size and number. A nonfunctional version of MFE2 did not restore normal peroxisome morphology to mfe2-KO cells, indicating that their phenotype is not due to the absence of MFE2. mfe2-KO cells contain higher amounts of beta-oxidation enzymes than do wild-type cells. We also show that increasing the level of the beta-oxidation enzyme thiolase results in enlarged peroxisomes. Our results implicate peroxisomal beta oxidation in the control of peroxisome size and number in yeast. PMID- 10787423 TI - The phosphotyrosyl phosphatase activator gene is a novel p53 target gene. AB - The minimal promoter of the phosphotyrosyl phosphatase activator (PTPA) gene, encoding a regulator of protein phosphatase 2A contains two yin-yang 1 (YY1) binding sites, positively regulating promoter activity. We now describe a role for p53 in the regulation of PTPA expression. Luciferase reporter assays in Saos 2 cells revealed that p53 could down-regulate PTPA promoter activity in a dose dependent manner, whereas four different p53 mutants could not. The p53 responsive region mapped to the minimal promoter. Overexpression of YY1 reverses the repressive effect of p53, suggesting a functional antagonism between p53 and YY1. The latter does not involve competition for YY1 binding, but rather direct control of YY1 function. Inhibition of PTPA expression by endogenous p53 was demonstrated in UVB-irradiated HepG2 cells, both on the mRNA and protein level. Also basal PTPA levels are higher in p53-negative (Saos-2) versus p53-positive (HepG2, U2OS) cells, suggesting "latent" p53 can control PTPA expression as well. The higher PTPA levels in U2OS cells, programmed to overexpress constitutively a dominant-negative p53 mutant, corroborate this finding. Thus, PTPA expression is negatively regulated by p53 in normal conditions and in conditions where p53 is up-regulated, via an as yet unknown mechanism involving the negative control of YY1. PMID- 10787424 TI - Inhibition of spontaneous beta 2-adrenergic activation rescues beta 1-adrenergic contractile response in cardiomyocytes overexpressing beta 2-adrenoceptor. AB - Cardiac-specific overexpression of the human beta(2)-adrenergic receptor (AR) in transgenic mice (TG4) enhances basal cardiac function due to ligand-independent spontaneous beta(2)-AR activation. However, agonist-mediated stimulation of either beta(1)-AR or beta(2)-AR fails to further enhance contractility in TG4 ventricular myocytes. Although the lack of beta(2)-AR response has been ascribed to an efficient coupling of the receptor to pertussis toxin-sensitive G(i) proteins in addition to G(s), the contractile response to beta(1)-AR stimulation by norepinephrine and an alpha(1)-adrenergic antagonist prazosin is not restored by pertussis toxin treatment despite a G(i) protein elevation of 1.7-fold in TG4 hearts. Since beta-adrenergic receptor kinase, betaARK1, activity remains unaltered, the unresponsiveness of beta(1)-AR is not caused by betaARK1-mediated receptor desensitization. In contrast, pre-incubation of cells with anti adrenergic reagents such as muscarinic receptor agonist, carbachol (10(-5)m), or a beta(2)-AR inverse agonist, ICI 118,551 (5 x 10(-7)m), to abolish spontaneous beta(2)-AR signaling, both reduce the base-line cAMP and contractility and, surprisingly, restore the beta(1)-AR contractile response. The "rescued" contractile response is completely reversed by a beta(1)-AR antagonist, CGP 20712A. Furthermore, these results from the transgenic animals are corroborated by in vitro acute gene manipulation in cultured wild type adult mouse ventricular myocytes. Adenovirus-directed overexpression of the human beta(2)-AR results in elevated base-line cAMP and contraction associated with a marked attenuation of beta(1)-AR response; carbachol pretreatment fully revives the diminished beta(1) AR contractile response. Thus, we conclude that constitutive beta(2)-AR activation induces a heterologous desensitization of beta(1)-ARs independent of betaARK1 and G(i) proteins; suppression of the constitutive beta(2)-AR signaling by either a beta(2)-AR inverse agonist or stimulation of the muscarinic receptor rescues the beta(1)-ARs from desensitization, permitting agonist-induced contractile response. PMID- 10787425 TI - Human ERK1 induces filamentous growth and cell wall remodeling pathways in Saccharomyces cerevisiae. AB - Expression of an activated extracellular signal-regulated kinase 1 (ERK1) construct in yeast cells was used to examine the conservation of function among mitogen-activated protein (MAP) kinases. Sequence alignment of the human MAP kinase ERK1 with all Saccharomyces cerevisiae kinases reveals a particularly strong kinship with Kss1p (invasive growth promoting MAP kinase), Fus3p (pheromone response MAP/ERK kinase), and Mpk1p (cell wall remodeling MAP kinase). A fusion protein of constitutively active human MAP/ERK kinase 1 (MEK) and human ERK1 was introduced under regulated expression into yeast cells. The fusion protein (MEK/ERK) induced a filamentation response element promoter and led to a growth retardation effect concomitant with a morphological change resulting in elongated cells, bipolar budding, and multicell chains. Induction of filamentous growth was also observed for diploid cells following MEK/ERK expression in liquid culture. Neither haploids nor diploids, however, showed marked penetration of agar medium. These effects could be triggered by either moderate MEK/ERK expression at 37 degrees C or by high level MEK/ERK expression at 30 degrees C. The combination of high level MEK/ERK expression and 37 degrees C resulted in cell death. The deleterious effects of MEK/ERK expression and high temperature were significantly mitigated by 1 m sorbitol, which also enhanced the filamentous phenotype. MEK/ERK was able to constitutively activate a cell wall maintenance reporter gene, suggesting misregulation of this pathway. In contrast, MEK/ERK effectively blocked expression from a pheromone-responsive element promoter and inhibited mating. These results are consistent with MEK/ERK promoting filamentous growth and altering the cell wall through its ability to partially mimic Kss1p and stimulate a pathway normally controlled by Mpk1p, while appearing to inhibit the normal functioning of the structurally related yeast MAP kinase Fus3p. PMID- 10787426 TI - Activation of the luteinizing hormone beta promoter by gonadotropin-releasing hormone requires c-Jun NH2-terminal protein kinase. AB - Regulation of the mitogen-activated protein kinase (MAPK) family by gonadotropin releasing hormone (GnRH) in the gonadotrope cell line LbetaT2 was investigated. Treatment with gonadotropin-releasing hormone agonist (GnRHa) activates extracellular signal-regulated kinase (ERK) and c-Jun NH(2)-terminal kinase (JNK). Activation of ERK by GnRHa occurred within 5 min, and declined thereafter, whereas activation of JNK by GnRHa occurred with a different time frame, i.e. it was detectable at 5 min, reached a plateau at 30 min, and declined thereafter. GnRHa-induced ERK activation was dependent on protein kinase C or extracellular and intracellular Ca(2+), whereas GnRHa-induced JNK activation was not dependent on protein kinase C or on extracellular or intracellular Ca(2+). To determine whether a mitogen-activated protein kinase family cascade regulates rat luteinizing hormone beta (LHbeta) promoter activity, we transfected the rat LHbeta (-156 to +7)-luciferase construct into LbetaT2 cells. GnRH activated the rat LHbeta promoter activity in a time-dependent manner. Neither treatment with a mitogen-activated protein kinase/ERK kinase (MEK) inhibitor, PD98059, nor cotransfection with a catalytically inactive form of a mitogen-activated protein kinase construct inhibited the induction of the rat LHbeta promoter by GnRH. Furthermore, cotransfection with a dominant negative Ets had no effect on the response of the rat LHbeta promoter to GnRH. On the other hand, cotransfection with either dominant negative JNK or dominant negative c-Jun significantly inhibited the induction of the rat LHbeta promoter by GnRH. In addition, GnRH did not induce either the rat LHbeta promoter activity in LbetaT2 cells transfected stably with dominant negative c-Jun. These results suggest that GnRHa differentially activates ERK and JNK, and a JNK cascade is necessary to elicit the rat LHbeta promoter activity in a c-Jun-dependent mechanism in LbetaT2 cells. PMID- 10787427 TI - Hepatic lipase deficiency decreases the selective uptake of HDL-cholesteryl esters in vivo. AB - Recent in vitro studies have provided evidence that hepatic lipase (HL) facilitates the selective uptake of HDL cholesteryl esters (CE), but the in vivo physiological relevance of this process has not been demonstrated. To evaluate the role that HL plays in facilitating the selective uptake of HDL-CE in vivo, we studied the metabolism of [(3)H]CEt, (125)I-labeled apolipoprotein (apo) A-I, and (131)I-labeled apoA-II-labeled HDL in HL-deficient mice. Kinetic analysis revealed similar catabolism of (125)I-labeled apoA-I (as well as (131)I-labeled apoA-II) in C57BL controls and HL deficient mice, with fractional catabolic rates (FCR) of 2.17 +/- 0.15 and 2.16 +/- 0.11 d(-)(1) (2.59 +/- 0.14 and 2.67 +/- 0.13 d(-)(1), respectively). In contrast, despite similar hepatic scavenger receptor BI expression, HL-deficient mice had delayed clearance of [(3)H]CEt compared to controls (FCR = 3.66 +/- 0.29 and 4.41 +/- 0.18 d(-)(1), P < 0.05). The hepatic accumulation of [(3)H]CEt in HL-deficient mice (62.3 +/- 2.1% of total) was significantly less than in controls (72.7 +/- 3.0%), while the [(3)H]CEt remaining in the plasma compartment increased (20.7 +/- 1.8% and 12.6 +/- 0.5%) (P < 0.05, all). In summary, HL deficiency does not alter the catabolism of apoA I and apoA-II but decreases the hepatic uptake and the plasma clearance of HDL CE. These data establish for the first time an important role for HL in facilitating the selective uptake of HDL-CE in vivo. PMID- 10787428 TI - Localization of the murine Niemann-Pick C1 protein to two distinct intracellular compartments. AB - Niemann-Pick type C (NPC) disease is characterized by an accumulation of cholesterol and other lipids in the lysosomal compartment. In this report, we use subcellular fractionation and microscopy to determine the localization of the murine Niemann-Pick C1 (NPC1) protein. Fractionation of mouse liver homogenates indicates that some NPC1 cosediments with lysosome-associated membrane protein 1 (LAMP1)-containing membranes. However, a significant amount of NPC1 is also found in membranes that do not contain LAMP1. Moreover, fractionation of liver membranes and fibroblasts in the presence of a nonionic detergent showed that a fraction of NPC1 cosediments with caveolin-1 in rafts. Immunofluorescence microscopy of cultured mouse fibroblasts showed that NPC1 is found in two morphologically distinct structures. The first population is characterized by large punctate structures that do not colocalize with major organelle protein markers, but do colocalize with filipin and a small fraction of caveolin-1. Examination of these large NPC1-containing compartments using electron microscopy shows that these structures contain extensive internal membranes. The second population is represented by smaller, more diffuse structures, a fraction of which colocalize with LAMP1-positive compartments. Incubation of fibroblasts with low density lipoprotein (LDL) increases colocalization of NPC1 with LAMP1 containing compartments. This colocalization can be further enhanced by treating fibroblasts with progesterone or chloroquine. The results indicate that NPC1 is associated with an unique vesicular compartment enriched with cholesterol and containing caveolin-1, and that NPC1 cycles to LAMP1-positive compartments, presumably to facilitate the processing of LDL-derived cholesterol. PMID- 10787429 TI - Induction of CD36 expression by oxidized LDL and IL-4 by a common signaling pathway dependent on protein kinase C and PPAR-gamma. AB - CD36, a class B scavenger receptor, is a macrophage receptor for oxidized low density lipoprotein (OxLDL) and may play a critical role in atherosclerotic foam cell formation. We have previously demonstrated that OxLDL, macrophage-colony stimulating factor (M-CSF), and interleukin-4 (IL-4) enhanced expression of CD36. The effect of OxLDL on CD36 is due, in part, to its ability to activate the transcription factor, PPAR-gamma (peroxisome proliferator activated receptor gamma). Other PPAR-gamma ligands (15-deoxyDelta(12,14) prostaglandin J(2) (15d PGJ(2)) and the thiazolidinedione class of antidiabetic drugs) also increase CD36 expression. We have now evaluated signaling pathways involved in the induction of CD36. Treatment of RAW264.7 cells (a murine macrophage cell line) with protein kinase C (PKC) activators (diacylglycerol and ingenol) up-regulated CD36 mRNA expression. Specific inhibitors of PKC reduced CD36 expression in a time dependent manner, while protein kinase A (PKA) and cyclic AMP agonists had no effect on CD36 mRNA expression. PKC inhibitors reduced basal expression of CD36 and blocked induction of CD36 mRNA by 15d-PGJ(2), OxLDL and IL-4. In addition, PKC inhibitors decreased both PPAR-gamma mRNA and protein expression and blocked induction of CD36 protein surface expression by OxLDL and 15d-PGJ(2) in human monocytes, as determined by FACS. 15d-PGJ(2) had no effect on translocation of PKC-alpha from the cytosol to the plasma membrane. These results demonstrate that two divergent physiological or pathophysiological agonists utilize a common pathway to up-regulate of CD36 gene expression. This pathway involves initial activation of PKC with subsequent PPAR-gamma activation. Defining these signaling pathways is critical for understanding and modulating expression of this scavenger receptor pathway. PMID- 10787430 TI - Modulation of plasma lipid levels and cholesterol kinetics by phytosterol versus phytostanol esters. AB - It has been suggested that phytosterol and phytostanol esters possess similar cholesterol-lowering properties, however, whether mechanisms responsible are identical has not been addressed. To address this question, cholesterol plasma levels, absorption, biosynthesis, and turnover were measured in 15 hypercholesterolemic subjects consuming prepared diets each over 21 d using a cross-over design. Diets contained either i) margarine (M), ii) margarine with phytosterol esters (MSE) (1.84 g/d), or iii) margarine with phytostanol esters (MSA) (1.84 g/d). Cholesterol absorption was measured using the ratio of [(13)C]cholesterol(oral):D(7)-cholesterol(IV); biosynthesis using D incorporation from D(2)O and turnover by D(7)-cholesterol(IV) decay rates. Plasma total cholesterol level at d 21/22 was lower (P < 0. 05) for MSE (13.4%) but not MSA (10.2%) versus M (6.0%) diets. Plasma low density lipoprotein-cholesterol (LDL-C) mean reductions at d 21/22 were larger (P < 0.05) for MSE (12.9%) and MSA (7.9%) compared with M (3.9%). Plasma TG and high density lipoprotein-cholesterol (HDL C) levels did not differ across diets. Cholesterol absorption was reduced (P < 0.05) 36.2 and 25.9% at d 21 for MSE and MSA versus M, while cholesterol biosynthesis was reciprocally increased (P < 0.05) 53.3 and 37.8% for MSE and MSA versus M, respectively. Cholesterol turnover was not influenced by diet. These data indicate that plant sterol and stanol esters differentially lower circulating total and LDL cholesterol levels by suppression of cholesterol absorption in hypercholesterolemic subjects. PMID- 10787431 TI - Plasma kinetics of apoC-III and apoE in normolipidemic and hypertriglyceridemic subjects. AB - Apolipoprotein (apo) C-III and apoE play a central role in controlling the plasma metabolism of triglyceride-rich lipoproteins (TRL). We have investigated the plasma kinetics of total, very low density lipoprotein (VLDL) and high density lipoprotein (HDL) apoC-III and apoE in normolipidemic (NL) (n = 5), hypertriglyceridemic (HTG, n = 5), and Type III hyperlipoproteinemic (n = 2) individuals. Apolipoprotein kinetics were investigated using a primed constant (12 h) infusion of deuterium-labeled leucine. HTG and Type III patients had reduced rates of VLDL apoB-100 catabolism and no evidence of VLDL apoB-100 overproduction. Elevated (3- to 12-fold) total plasma and VLDL apoC-III levels in HTG and Type III patients, although associated with reduced apoC-III catabolism (i.e., increased residence times (RTs)), were mainly due to increased apoC-III production (plasma apoC-III transport rates (TRs, mean +/- SEM): (NL) 2.05 +/- 0.22 (HTG) 4.90 +/- 0.81 (P < 0.01), and (Type III) 8.78 mg. kg(-)(1). d(-)(1); VLDL apoC-III TRs: (NL) 1.35 +/- 0. 23 (HTG) 5.35 +/- 0.85 (P < 0.01), and (Type III) 7.40 mg. kg(-)(1). d(-)(1)). Elevated total plasma and VLDL apoE levels in HTG (2- and 6-fold, respectively) and in Type III (9- and 43-fold) patients were associated with increased VLDL apoE RTs (0.21 +/- 0.02, 0.46 +/- 0. 05 (P < 0.01), and 1.21 days, NL vs. HTG vs. Type III, respectively), as well as significantly increased apoE TRs (plasma: (NL) 2.94 +/- 0.78 (HTG) 5.80 +/- 0.59 (P < 0.01) and (Type III) 11.80 mg. kg(-)(1). d(-)(1); VLDL: (NL) 1.59 +/- 0.18 (HTG) 4.52 +/- 0.61 (P < 0.01) and (Type III) 11.95 mg. kg(-)(1). d(-)(1)). These results demonstrate that hypertriglyceridemic patients, having reduced VLDL apoB 100 catabolism (including patients with type III hyperlipoproteinemia) are characterized by overproduction of plasma and VLDL apoC-III and apoE. PMID- 10787432 TI - Effect of dietary fish oil on the sensitivity of hepatic lipid metabolism to regulation by insulin. AB - The contribution of dietary fat content and type to changes in the sensitivity of hepatic lipid metabolism to insulin was studied in primary hepatocyte cultures from donor rats maintained on a low-fat diet (LF), or on diets enriched in olive oil (OO) or fish oil (FO). The higher rate of fatty acid oxidation in hepatocytes from the FO-fed group was resistant to the inhibitory effects of insulin observed in hepatocytes from the other groups. Insulin stimulation of fatty acid incorporation into triglyceride (TG) was also less pronounced in hepatocytes from the FO-fed group than in those from the OO-fed group but there was no difference in the stimulatory effect of insulin on fatty acid incorporation into phospholipid (PL) in these two groups. In the case of fatty acid incorporation into both PL and TG, hepatocytes from the LF group were refractory to stimulation by insulin. At each concentration of insulin, hepatocytes from the FO-fed group secreted less very low density lipoprotein (VLDL) TG than those from the other groups. However, the absolute suppression of VLDL TG secretion by insulin was similar irrespective of the diet of the donor animals.We conclude that chronic consumption of a particular type of dietary fat does not affect the insulin sensitivity of the major pathways of hepatic lipid metabolism in a consistent manner. PMID- 10787433 TI - Influence of LDL apheresis on LDL subtypes in patients with coronary heart disease and severe hyperlipoproteinemia. AB - Epidemiologic studies and in vitro experiments indicate that low density lipoprotein (LDL) subtypes differ concerning their atherogenic potential. Small, dense LDL are more atherogenic than large, buoyant LDL. LDL apheresis is a potent therapeutic modality to lower elevated LDL-cholesterol. It is unknown whether such therapy induces a shift in the LDL subtype distribution. In this study we evaluated the influence of LDL apheresis on the LDL subtype distribution in patients with CHD and familial hypercholesterolemia (FH, n = 22), combined hyperlipidemia (CHLP, n = 6), or Lp[a]-hyperlipoproteinemia (Lp[a]-HLP, n = 4) regularly treated by LDL apheresis (immunoadsorption (n = 14), HELP apheresis (n = 8), dextran sulfate adsorption (n = 7), cascade filtration (n = 3)). On the basis of 6 LDL subfractions (d 1.020;-1.057 g/mL) isolated by density gradient ultracentrifugation the LDL-density profile was determined in each patient before and after apheresis. There was a relative increase of LDL-subfractions 1, 2, and 3 (P < 0.01, P < 0. 05, and P < 0.01, respectively) and a concomitant decrease of LDL subfractions 5 and 6 (P < 0.05) after apheresis. Subgroup analysis indicates that the degree of the small, dense LDL reduction was much more prominent in patients with CHLP compared to patients with FH or Lp[a]-HLP, whereas the type of apheresis technique had no effect. The extent of small, dense LDL reduction correlated with the preapheresis concentrations of small, dense LDL and triglycerides but not with the extent of triglyceride reduction.We conclude that LDL apheresis not only decreases LDL mass, but also improves LDL-density profile, particularly in patients with CHLP. PMID- 10787434 TI - Two novel mutations in the lipoprotein lipase gene in a family with marked hypertriglyceridemia in heterozygous carriers. Potential interaction with the polymorphic marker D1S104 on chromosome 1q21-q23. AB - Two novel mutations in the lipoprotein lipase (LPL) gene are described in an Austrian family: a splice site mutation in intron 1 (3 bp deletion of nucleotides -2 to -4) which results in skipping of exon 2, and a missense mutation in exon 5 which causes an asparagine for histidine substitution in codon 183 and complete loss of enzyme activity. A 5-year-old boy who exhibited all the clinical features of primary hyperchylomicronemia was a compound heterozygote for these two mutations. Nine other family members were investigated: seven were heterozygotes for the splice site mutation, one was a heterozygote for the missense mutation, and one had two wild-type alleles of the LPL gene. LPL activity in the post heparin plasma of the heterozygotes was reduced to 49;-79% of the mean observed in normal individuals. Two of the heterozygotes had extremely high plasma triglyceride levels; in three of the other heterozygotes the plasma triglycerides were also elevated. As plasma triglycerides in carriers of one defective LPL allele can be normal or elevated, the heterozygotes of this family have been studied for a possible additional cause of the expression of hypertriglyceridemia in these subjects. Body mass index, insulin resistance, mutations in other candidate genes (Asn291Ser and Asp9Asn in the LPL gene, apoE isoforms, polymorphisms in the apoA-II gene and in the apoAI-CIII-AIV gene cluster, and in the IRS-1 gene) could be ruled out as possible factors contributing to the expression of hypertriglyceridemia in this family. A linkage analysis using the allelic marker D1S104 on chromosome 1q21;-q23 suggested that a gene in this region could play a role in the expression of hypertriglyceridemia in the heterozygous carriers of this family, but the evidence was not sufficiently strong to prove this assumption. Nevertheless, this polymorphic marker seems to be a good candidate for further studies. PMID- 10787435 TI - Evidence against the peroxisome proliferator-activated receptor alpha (PPARalpha) as the mediator for polyunsaturated fatty acid suppression of hepatic L-pyruvate kinase gene transcription. AB - The glycolytic enzyme, L-pyruvate kinase (L-PK), plays an important role in hepatic glucose metabolism. Insulin and glucose induce L-PK gene expression, while glucagon and polyunsaturated fatty acids (PUFA) inhibit L-PK gene expression. We have been interested in defining the PUFA regulation of L-PK. The cis-regulatory target for PUFA action includes an imperfect direct repeat (DR1) that binds HNF-4. HNF4 plays an ancillary role in the insulin/glucose-mediated transactivation of the L-PK gene. Because the fatty acid-activated nuclear receptor, peroxisome proliferator-activated receptor (PPARalpha), binds DR1-like elements and has been reported to interfere with HNF4 action, we examined the role PPARalpha plays in the regulation of L-PK gene transcription. Feeding rats either fish oil or the potent PPARalpha activator, WY14,643, suppressed rat hepatic L-PK mRNA and gene transcription. The PPARalpha-null mouse was used to evaluate the role of the PPARalpha in hepatic transcriptional control of L-PK. While WY14,643 control of L-PK gene expression required the PPARalpha, PUFA regulation of L-PK gene expression was independent of the PPARalpha. Transfection studies in cultured primary hepatocytes localized the cis-regulatory target for WY14,643/PPARalpha action to the L-PK HNF4 binding site. However, PPARalpha/RXRalpha heterodimers did not bind this region. Although both WY14,643 and PUFA suppress L-PK gene transcription through the same element, PUFA regulation of L-PK does not require the PPARalpha and PPARalpha/RXRalpha does not bind the L-PK promoter. These studies suggest that other intermediary factors are involved in both the PUFA and PPARalpha regulation of L-PK gene transcription. PMID- 10787436 TI - Relationship between structure and biochemical phenotype of lecithin:cholesterol acyltransferase (LCAT) mutants causing fish-eye disease. AB - In order to test the hypothesis that fish-eye disease (FED) is due to a deficient activation of lecithin:cholesterol acyltransferase (LCAT) by its co-factor apolipoprotein (apo) A-I, we overexpressed the natural mutants T123I, N131D, N391S, and other engineered mutants in Cos-1 cells. Esterase activity was measured on a monomeric phospholipid enelogue, phospholipase A(2) activity was measured on reconstituted high density lipoprotein (HDL), and acyltransferase activity was measured both on rHDL and on low density lipoprotein (LDL). The natural FED mutants have decreased phospholipase A(2) activity on rHDL, which accounts for the decreased acyltransferase activity previously reported. All mutants engineered at positions 131 and 391 had decreased esterase activity on a monomeric substrate and decreased acyltransferase activity on LDL. In contrast, mutations at position 123 preserved these activities and specifically decreased phospholipase A(2) and acyltransferase activites on rHDL. Mutations of hydrophilic residues in amphipathic helices alpha 3;-4 and alpha His to an alanine did not affect the mutants' activity on rHDL. Based upon the 3D model built for human LCAT, we designed a new mutant F382A, which had a biochemical phenotype similar to the natural T123I FED mutant. These data suggest that residues T123 and F382, located N-terminal of helices alpha 3-4 and alpha His, contribute specifically to the interaction of LCAT with HDL and possibly with its co-factor apoA-I. Residues N131 and N391 seem critical for the optimal orientation of the two amphipathic helices necessary for the recognition of a lipoprotein substrate by the enzyme. PMID- 10787437 TI - Multiple mechanisms, independent of sterol regulatory element binding proteins, regulate low density lipoprotein gene transcription. AB - Transcription of the LDL receptor gene is markedly enhanced in the Jurkat T cell line by stimulation with the combination of the phorbol ester phorbol 12 myristate 13-acetate (PMA) and the protein synthesis inhibitor cycloheximide (CHX). The DNA sequences necessary for this response were identified by analysis of Jurkat T cells permanently transfected with reporter gene expression vectors containing fragments of the LDL receptor promoter extending from 68 bp to 1472 bp 5' of the major transcription start site. The magnitude of the response of this array of promoter fragments to stimulation with PMA and CHX was similar to that previously observed with a approximately 6.5 kb promoter fragment. However, the various promoter fragments differed with regard to the role of the sterol regulatory element-1 (SRE-1) sequence. Thus, whereas a 142 bp promoter mediated transcription stimulated by PMA and CHX independently of SRE-1, a shorter 115 bp promoter was absolutely dependent on SRE-1. Furthermore, internal deletion of promoter sequences from -142 bp to -113 bp from longer promoter constructs in which the SRE-1 was mutated prevented the induction of transcription by PMA and CHX. Electrophoretic mobility shift assays (EMSAs) demonstrated sequence specific, stimulus-independent binding by Jurkat nuclear proteins to the novel response element mapped between -142 and -115. Even though the minimal 115 bp or 68 bp promoter fragment required an intact SRE-1 to respond to PMA and CHX, transcriptional induction persisted when nuclear levels of sterol regulatory element binding proteins (SREBPs) were made undetectable by culture in suppressive sterols. Taken together, these data indicate that non-sterol stimuli such as the combination of PMA and CHX induce LDL receptor gene transcription through at least two distinct promoter elements, neither of which requires the presence of SREBPs. However, the element proximal to the transcription start site is dependent on the SRE-1. PMID- 10787438 TI - Transport of heptafluorostearate across model membranes. Membrane transport of long-chain fatty acid anions I. AB - Heptafluorostearic acid, an isogeometric derivative of stearic acid, has a pK(a) value of about 0.5. To evaluate the suitability of heptafluorostearate as model compound for anions of long-chain fatty acids in membrane transport, monolayer and liposome studies were performed with lipid mixtures containing phospholipids; cholesterol-heptafluorostearate or stearate (100:40:20 molar ratios). Transfer of heptafluorostearate and stearate from liposomes to bovine serum albumin (BSA) was followed by measuring the intrinsic fluorescence of BSA. The percentage of heptafluorostearate, equivalent to the amount placed in their outer monolayer, transferred from liposomes (120;-130 nm diameter) to BSA was 55.7 +/- 3.7% within 10 min at 25 degrees C and 55 +/- 2% within 5 min at 37 degrees C. Slow transfer of 22.7 +/- 2.5% of heptafluorostearate at 25 degrees C followed with a half-life of 2.3 +/- 0.4 h and of 20 +/- 4% at 37 degrees C with a half-life of 0.9 +/- 0.1 h until the final equilibrium distributions between BSA and liposomes were reached, 79 +/- 6% to 21 +/- 5% at 25 degrees C and 75 +/- 5% to 25 +/- 4% at 37 degrees C. The pseudounimolecular rate constants for flip-flop of heptafluorostearate equal k(FF,25) = 0.24 +/- 0.05 h(-) and k(FF,37) = 0.6 +/- 0.1 h(-), respectively. By comparison, transfer of stearate required only 3 min to reach equilibrium distribution. The difference between heptafluorostearate and stearate may be explained by a rapid flip-flop movement of the un-ionized fatty acids which exist in different concentrations in accordance with their pK(a) values. Half-life of flip-flop of heptafluorostearate makes it suitable to study mediated membrane transport of long-chain fatty acid anions. PMID- 10787439 TI - Sterol carrier protein-2 expression alters phospholipid content and fatty acyl composition in L-cell fibroblasts. AB - The effects sterol carrier protein-2 (SCP-2) expression on L-cell phospholipid levels and fatty acyl composition was assessed using L-cells transfected with the murine cDNA encoding for either the 15 kDa proSCP-2 or 13.2 kDa SCP-2. Expression of these proteins reduced total phospholipid mass (nmol/mg protein) by 24% and reduced the cholesterol to phospholipid ratio 60 and 28%, respectively. In 15 kDa proSCP-2 expressing cells, individual phospholipid class masses, excluding sphingomyelin (CerPCho), were reduced as follows: phosphatidylinositol (PtdIns) and phosphatidylserine (PtdSer) >> ethanolamine glycerophospholipid (EtnGpl) > choline glycerophospholipid (ChoGpl). Furthermore, ethanolamine plasmalogen mass was decreased 25%, while choline plasmalogen mass was elevated 30% in 15 kDa proSCP-2 expressing cells. In 13.2 kDa SCP-2 expressing cells, phospholipid class mass was decreased as follows: PtdIns and PtdSer >> ChoGpl. These changes in phospholipid mass resulted in altered cellular phospholipid composition. Expression of either protein differentially altered the type of fatty acid esterified onto the phospholipids. These effects included a greater proportion of polyunsaturated fatty acids and a reduction in saturated fatty acids, although 15 kDa proSCP-2 expression had a more robust effect on these parameters than did 13.2 kDa SCP-2 expression. In summary, expression of SCP-2 reduced individual phospholipid class mass, except for CerPCho, and altered the fatty acid composition of each phospholipid class examined. These results clearly demonstrate that SCP-2 expression altered basal phospholipid levels, suggesting that SCP-2 can alter the function of endoplasmic reticulum phospholipid synthetic enzymes. PMID- 10787440 TI - Dimorphic expression of cerebrosides in the mycopathogen Sporothrix schenckii. AB - Major neutral glycosphingolipid components were extracted from Sporothrix schenckii, a dimorphic fungus exhibiting a hyphal saprophytic phase and a yeast parasitic phase responsible for chronic mycotic infections in mammalian hosts. These components, one from the mycelial form and two from the yeast form, were purified and their structures were elucidated by (1)H nuclear magnetic resonance (NMR) spectroscopy, electrospray ionization mass spectrometry (ESI-MS), and tandem ESI-MS/MS. All three were characterized as cerebrosides (monohexosylceramides) containing (4E, 8E)-9-methyl-4,8-sphingadienine as the long-chain base attached to N-2'-hydroxyoctadecanoate and N-2'-hydroxy-(E) Delta(3)-octadecenoate as the fatty acyl components. However, while the mycelial form expressed only beta-glucopyranosylceramide, the yeast form expressed both beta-gluco- and beta-galactopyranosylceramides in approximately equal amounts. In addition, while the glucosylceramides of both mycelial and yeast forms had similar proportions of saturated and (E)-Delta(3) unsaturated 2-hydroxy fatty acid, the galactocerebroside of the yeast form had significantly higher levels of (E)-Delta(3) unsaturation. The differences in cerebroside hexose structure represent a novel type of glycosphingolipid dimorphism not previously reported in fungi. Possible implications of these findings with respect to regulation of morphological transitions in S. schenckii and other dimorphic fungi are discussed. PMID- 10787441 TI - Regulation of acetylated low density lipoprotein uptake in macrophages by pertussis toxin-sensitive G proteins. AB - Class A scavenger receptors (SR-A) mediate the uptake of modified low density lipoprotein (LDL) by macrophages. Although not typically associated with the activation of intracellular signaling cascades, results with peritoneal macrophages indicate that the SR-A ligand acetylated LDL (AcLDL) promotes activation of cytosolic kinases and phospholipases. These signaling responses were blocked by the treatment of cells with pertussis toxin (PTX) indicating that SR-A activates G(i/o)-linked signaling pathways. The functional significance of SR-A-mediated G(i/o) activation is not clear. In this study, we investigated the potential role of G(i/o) activation in regulating SR-A-mediated lipoprotein uptake. Treatment of mouse peritoneal macrophages with PTX decreased association of fluorescently labeled AcLDL with cells. This inhibition was dependent on the catalytic activity of the toxin confirming that the decrease in AcLDL uptake involved inhibiting G(i/o) activation. In contrast to the inhibitory effect on AcLDL uptake, PTX treatment did not alter beta-VLDL-induced cholesterol esterification or deposition of cholesterol. The ability of polyinosine to completely inhibit AcLDL uptake, and the lack of PTX effect on beta-VLDL uptake, demonstrated that the inhibitory effect is specific for SR-A and not the result of non-specific effects on lipoprotein metabolism. Despite having an effect on an SR-A-mediated lipoprotein uptake, there was no change in the relative abundance of SR-A protein after PTX treatment. These results demonstrate that activation of a PTX-sensitive G protein is involved in a feedback process that positively regulates SR-A function. PMID- 10787442 TI - Involvement of the peroxisome proliferator-activated receptor alpha in regulating long-chain acyl-CoA thioesterases. AB - Long-chain acyl-CoA thioesterases catalyze the hydrolysis of acyl-CoAs to the corresponding free fatty acid and CoA. We recently cloned four members of a novel multi-gene family of peroxisome proliferator-induced genes encoding cytosolic (CTE-I), mitochondrial (MTE-I), and peroxisomal (PTE-Ia and PTE-Ib) acyl-CoA thioesterases (Hunt et al. 1999. J. Biol. Chem. 274: 34317-34326). As the peroxisome proliferator-activated receptor alpha (PPARalpha) plays a central role in regulating genes involved in lipid metabolism, we examined the involvement of this receptor in regulation of the thioesterases, particularly CTE-I and MTE-I. Northern blot analysis shows that the induction of these thioesterases by clofibrate is mediated through a strictly PPARalpha-dependent mechanism. All four acyl-CoA thioesterases are induced at mRNA level by fasting and using PPARalpha null mice, it is evident that the increase in CTE-I due to fasting is mainly independent of the PPARalpha in liver and heart. The CTE-I gene responds rapidly to fasting, with induction of mRNA and protein evident after 6 h. This fasting effect is rapidly reversible, with CTE-I mRNA returning almost to control levels after 3 h refeeding, and being further repressed to 20% of control after 9 h refeeding. Although CTE-I mRNA shows a low basal expression in liver, it can be suppressed 90% by feeding a fat-free diet. These data demonstrate that the nutritional regulation of the thioesterases involves the PPARalpha and other signaling pathways responsible for activation and repression. Putative physiological functions for the acyl-CoA thioesterases are discussed. PMID- 10787443 TI - Oxidized phospholipids, linked to apolipoprotein B of oxidized LDL, are ligands for macrophage scavenger receptors. AB - Previous studies have shown that macrophage receptors for oxidized LDL (OxLDL) recognize both the lipid and protein moieties, and that a monoclonal antibody against OxLDL, EO6, also recognizes both species. The present studies show directly that during LDL oxidation phospholipids become covalently attached to apolipoprotein B (apoB). After exhaustive extraction of lipids, apoB of native LDL contained 4 +/- 3 moles of phosphorus/mole protein. In contrast, apoB of OxLDL contained approximately 75 moles of phosphorus/mole protein. Saponification of this apoB released phosphorus, choline, and saturated fatty acids in a molar ratio of 1.0:0.98:0.84. When LDL was reductively methylated prior to oxidation, the amount of phospholipid covalently bound was reduced by about 80%, indicating that the phospholipids attach at lysine epsilon amino groups. Progressive decreases in the phospholipid associated with apoB of OxLDL decreased the ability of the protein to compete for binding to macrophage scavenger receptors and decreased its reactivity with antibody EO6. We postulate that some oxidized phospholipids containing fatty acid aldehydes at the sn-2 position bind to lysine residues of apoB while others remain unreacted within the lipid phase. This would account for the interchangeability of lipid and apolipoprotein of OxLDL with respect to receptor binding and antibody recognition. PMID- 10787444 TI - Hydrogenated fat consumption affects cholesterol synthesis in moderately hypercholesterolemic women. AB - To determine mechanisms by which hydrogenated fat influences plasma lipid levels, 14 women (65;-71 yrs with LDL-C >/= 130 mg. dl(-)(1)) consumed, for 5-week periods each, a baseline (BL) diet (39% kcal fat, 164 mg chol. 1000 kcal(-)(1)) and reduced fat diets (30% kcal) where two-thirds of the fat was either soybean oil (SO), low trans squeeze (SQM), medium trans tub (TM), or high trans stick (SM) margarines, or butter (BT). Plasma lipid levels were analyzed at the end of each phase. Fractional synthesis rates (FSR) in pools/day (p. d(-)(1)) and absolute synthesis rates (ASR) in grams/day (g. d(-)(1)) of free cholesterol (FC) were measured using the deuterium incorporation methodology. Plasma total (P < 0.01) and low density lipoprotein (P < 0.05) cholesterol levels increased with increasing degree of hydrogenation or saturated fat intake. High density lipoprotein cholesterol levels (P < 0.05) were lowest on the SM diet when compared to the BT diet. Low trans SQM (0.081 +/- 0.019 p. d(-)(1)) and medium trans TM (0.086 +/- 0.029 p. d(-)(1)) diets elicited responses similar to the SO (0.078 +/- 0.024 p. d(-)(1)) diet, whereas high trans SM (0.053 +/- 0.029 p. d( )(1)) diet mimicked the BT (0.062 +/- 0.017 p. d(-)(1)) and high fat BL (0.053 +/ 0.023 p. d(-)(1)) diet in its suppression (P < 0.05) of FSR-FC. ASR-FC, which is an approximation of the daily production of newly synthesized cholesterol, showed a trend similar to the FSR-FC data. These results indicate that reduced synthesis is not responsible for the higher plasma TC levels seen with consumption of the SM, BT, and BL diets, and suggest that another mechanism, possibly impairment of the catabolic pathway of cholesterol, is involved. PMID- 10787445 TI - Plasma levels of 24S-hydroxycholesterol reflect the balance between cerebral production and hepatic metabolism and are inversely related to body surface. AB - We have previously presented evidence that most of the 24S-hydroxycholesterol present in the circulation originates from the brain and that most of the elimination of this oxysterol occurs in the liver. Plasma 24S-hydroxycholesterol levels decline by a factor of about 5 during the first decades of life. The concentration of the enzyme cholesterol 24S-hydroxylase in the brain is, however, about constant from the first year of life, and reduced enzyme levels thus cannot explain the decreasing plasma levels during infancy. In the present work we tested the hypothesis that the plasma levels of 24S-hydroxycholesterol may reflect the size of the brain relative to the capacity of the liver to eliminate the substance. It is shown here that the age-dependent changes in absolute as well as cholesterol-related plasma level of 24S-hydroxycholesterol closely follow the changes in the ratio between estimated brain weight and estimated liver volume. The size of the brain is increased only about 50% whereas the size of the liver is increased by about 6-fold after the age of 1 year. Liver volume is known to be highly correlated to body surface, and in accordance with this the absolute as well as the cholesterol-related plasma level of 24S-hydroxycholesterol was found to be highly inversely correlated to body surface in 77 healthy subjects of varying ages (r(2) = 0.74). Two chondrodystrophic dwarves with normal size of the brain but with markedly reduced body area had increased levels of 24S hydroxycholesterol when related to age but normal levels when related to body surface. It is concluded that the balance between cerebral production and hepatic metabolism is a critical determinant for plasma levels of 24S-hydroxycholesterol at different ages and that endocrinological factors are less important. The results are discussed in relation to the possibility to use 24S hydroxycholesterol in the circulation as a marker for cholesterol homeostasis in the brain. PMID- 10787446 TI - Enhancement of hydrolytic activity of sphingolipid ceramide N-deacylase in the aqueous-organic biphasic system. AB - Lysoglycosphingolipids were produced from glycosphingolipids by using sphingolipid ceramide N-deacylase, which cleaves the N-acyl linkage between fatty acids and sphingosine bases in various glycosphingolipids. The enzyme reaction was done in a biphasic media prepared with water;-immiscible organic solvent and aqueous buffer solution containing the enzyme. We investigated the effects of organic solvents and detergents on lysoglycosphingolipid production in the biphasic system. Among the organic solvents tested, n-butylbenzene, cumene, cyclodecane, cyclohexane, n-decane, diisopropylether, n-heptadecane, and methylcyclohexane promoted hydrolysis of GM1, whereas benzene, chloroform, ethyl acetate, and toluene inhibited GM1 hydrolysis. Hydrolysis of asialo GM1, GD1a, GalCer, and sulfatide was also enhanced by the addition of n-decane. The hydrolytic activity of the enzyme was enhanced by the addition of 0.8% sodium taurodeoxycholate or sodium cholate to the aqueous phase. The most effective hydrolysis of various glycosphingolipids by the enzyme was thus obtained in the aqueous-n-decane biphasic system containing 0.8% sodium taurodeoxycholate. Under this condition, the fatty acids released from GM1 by the action of the enzyme were trapped and diffused into the organic phase, while lysoGM1 remained in the aqueous phase. Thus the almost complete hydrolysis of GM1 was achieved using the biphasic system, while at most 70% of hydrolysis was obtained using normal aqueous media possibly due to the inhibition of hydrolysis reaction by accumulation of fatty acids in the reaction mixture. PMID- 10787447 TI - Controlling hypertension in the elderly. PMID- 10787448 TI - Bartter syndrome: an overview. AB - The term Bartter syndrome denotes a group of renal diseases which share a common denominator of hypokalaemia and metabolic alkalosis. The patch-clamp technique has made possible the analysis of single ion channels, improving our understanding of the molecular physiopathology of all the 'Bartter-like' syndromes. Genetic mapping of each defect has further clarified the mutations involved and the possible modes of inheritance. This improved understanding has opened new avenues for therapy, improving mortality and morbidity in these patients. Another group of illnesses, the 'pseudo-Bartter syndrome', may produce a hypokalaemic metabolic alkalosis without primary renal disease. The underlying illness needs to be identified and treated. PMID- 10787449 TI - The genetic basis of autosomal dominant familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is classically an autosomal recessive periodic inflammatory disease occurring in Mediterranean and Middle Eastern populations. It is caused by mutations affecting both alleles of MEFV, a gene that encodes pyrin (marenostrin), an uncharacterized neutrophil protein. Occasional reports of autosomal dominant FMF have often been discounted, on the basis that asymptomatic FMF carriers are common in certain populations, and give rise to pseudo-dominant inheritance. We performed comprehensive MEFV genotyping in five families in whom FMF appeared to be inherited dominantly. Transmission proved to be pseudo dominant in two cases, but true dominant inheritance of FMF with variable penetrance was supported by the genotyping results in the other three families. The disease in these cases was associated with heterozygosity for either pyrin DeltaM694 alone or the compound pyrin variant E148Q/M694I, the latter occurring in two unrelated families. Complete MEFV sequencing failed to identify any coding region abnormality in the other allele in any of these cases, and, in the largest kindred, single-allele disease transmission was further supported by analysis of silent single nucleotide polymorphisms, which proved that affected individuals had at least three different complementary alleles. Studies of two further unrelated British patients with FMF associated with simple heterozygosity for pyrin DeltaM694 were also consistent with autosomal dominant inheritance. The clinical features of dominantly inherited FMF were absolutely typical, including AA amyloidosis in a patient with pyrin DeltaM694. These findings extend the spectrum of FMF, and suggest that the methionine residue at position 694 makes a crucial contribution to pyrin's function, and that a 50% complement of normal pyrin activity does not prevent susceptibility to FMF. PMID- 10787450 TI - Clinical versus genetic diagnosis of familial Mediterranean fever. AB - The diagnosis of familial Mediterranean fever (FMF) has until recently been based on clinical signs alone. Discovery of the MEFV gene has enabled a molecular approach to diagnosis, which is already well established for diagnosing typical clinical forms of FMF. We evaluated the utility of this molecular approach in a large series of patients with various clinical presentations and ethnic origins. We looked for mutations in the MEFV gene in 303 unselected consecutive patients with a variable (from high to low) clinical suspicion of FMF. Two mutations were found in 133 patients (44%). In 22 patients (7%), the clinical diagnosis of FMF was unlikely according to the Tel Hashomer clinical criteria. Our results suggest that the spectrum of FMF-associated signs is broader than previously believed. Wider indications for genotyping should lead to more frequent diagnosis of FMF. PMID- 10787451 TI - Determinants of raised C-reactive protein concentration in type 1 diabetes. AB - As a marker of systemic inflammation, raised C-reactive protein (CRP) concentrations which are still within the normal range have been associated with an increased incidence of coronary heart disease (CHD) in non-diabetic subjects. This study aimed to establish potential determinants of raised CRP concentrations in type 1 diabetic patients. We used a sensitive assay to measure 'low-level' CRP concentrations in 167 type 1 patients (93M, 74F, median age 30 years, range 13 67). Stepwise multivariate analysis was used to relate these CRP levels to known cardiovascular risk factors and demographic data. Only six patients had established CHD (median CRP 3.34 mg/l vs. 0.83 mg/l, p=0.032). In subjects without overt CHD, multivariate analysis showed increases in subject age (p=0.0025), BMI (p=0.001) and HbA(1) (p=0.012) to be associated with a higher CRP concentration, as was female sex (p=0.026) and a history of CHD in a first-degree relative (p=0.018, n=57). The duration of diabetes, current smoking status, presence of microvascular complications, lipid status and presence of hypertension were unrelated. This study suggests that some of the risk factors associated with CHD in type 1 patients are also independently predictive of high CRP concentrations. The reasons for this, and whether intervention would prove useful, require further investigation. PMID- 10787452 TI - Massive hepatic fibrosis in Gaucher's disease: clinico-pathological and radiological features. AB - Hepatomegaly is frequent in patients with type 1 Gaucher's disease and is associated with infiltration of the liver with pathological macrophages. Most patients suffer no significant clinical consequences, but a few develop portal hypertension which may progress to parenchymal liver failure. We describe four patients with Gaucher's disease who have developed portal hypertension. We have reviewed their clinical histories and all available histological and radiological material. All had severe Gaucher's disease with multi-organ involvement, and had undergone splenectomy in childhood. Histologically, this advanced liver disease was characterized by a picture of extreme and advanced confluent fibrosis occupying the central region of the liver. This massive fibrosis is associated with characteristic radiological appearances. The liver histology in these cases is highly unusual and virtually unknown in other conditions. Our studies indicate that without specific treatment the liver disease is progressive and rapidly fatal. However, institution of enzyme replacement therapy with imiglucerase may have beneficial effects even when the condition is far advanced. PMID- 10787453 TI - Cryptococcal meningitis in non-HIV-infected patients. AB - There are few reports on cryptococcal meningitis in non-HIV-infected patients in subtropical areas. We reviewed 94 non-HIV-infected patients microbiologically diagnosed with cryptococcal meningitis and hospitalized at National Taiwan University Hospital, 1977-1996. Forty-two patients (44.7%) had underlying diseases. The main initial manifestations were headache (86.2%), vomiting (72.3%) and fever (69. 1%). The 30 patients with T-cell suppression had more acute illnesses (median duration of symptoms: 14 days vs. 29 days), less typical presentations of meningitis, and reduced inflammatory responses compared with the 64 without T cell suppression. There was no statistical difference between patients who received amphotericin B treatment for 10 weeks and those received amphotericin B with subsequent fluconazole treatment, in terms of mortality rate and recurrence rate. Seventy-five patients (79.8%) had satisfactory clinical responses, and two relapsed. Eighteen patients died (19.1%) and 10 of these died within 2 weeks of hospitalization. Patients in this series had outcomes comparable with those from temperate and even tropical countries with high percentages of immunocompetent hosts. Factors significantly associated with death were lymphoma, semicoma, leukocytosis, and initial high titres of cryptococcal antigen in cerebral spinal fluid (especially >/=1 : 512). On multivariate analysis, lymphoma and initial high cryptococcal antigen titres were independent predictors of mortality. PMID- 10787454 TI - Rationing renal replacement therapy to older patients--agreed guidelines are needed. PMID- 10787455 TI - Making good organizations better. PMID- 10787456 TI - Cardiovascular disease and technology: linking patients and professionals. PMID- 10787457 TI - As Canadian as hockey. PMID- 10787458 TI - Cardiology education and examination: evolution or revolution. PMID- 10787459 TI - The assessment of technical skills in a cardiology training program: is the ITER sufficient? AB - Diagnostic coronary angiography is an important tool in the assessment of heart disease. No objective data are available regarding the best method of training or testing a cardiology trainee's skill in the performance of this procedure. Training guidelines are based on the consensus opinions of experts in the field, and the assessment of proficiency is relegated to a generalized in-training evaluation report (ITER). However, is an ITER sufficient? The current modes of assessment are examined, with the question of whether a more comprehensive and valid assessment process is needed. PMID- 10787460 TI - Late coronary artery reperfusion: benefits and mechanisms. AB - In patients with acute myocardial infarction (AMI), coronary artery reperfusion beyond 4 to 6 h after the onset of symptoms is believed to have little or no effect on infarct size. However, benefits have been demonstrated even when reperfusion is performed late and without significant myocardial salvage. Late reperfusion prevents infarct expansion and left ventricular remodelling. The mechanism is multifactorial and partially related to salvage of small islets of still viable myocytes within the scar. Other benefits of late reperfusion are electrical stability and prevention of arrhythmias. The survival benefits of reperfusion are strongly dependent on the time after the onset of symptoms until treatment (better outcomes are obtained with earlier reperfusion). Based on the demonstration of a significant reduction in mortality, the current practice is to perform reperfusion up to 12 h after the onset of symptoms. The survival benefits of reperfusion performed 13 to 24 h after the onset of AMI are less certain; however, large AMIs, particularly anterior infarcts, have the maximum risk of remodelling. Thus, benefits of reperfusion on left ventricular remodelling, arrhythmogenicity and prevention of congestive heart failure justify reperfusion in this group of patients up to 24 h after the onset of symptoms. PMID- 10787461 TI - Factors predictive of oxygen consumption during the immediate postoperative period in open heart surgery. AB - BACKGROUND: Postoperative oxygen consumption (VO2) is critical during the recovery period that follows open heart surgery and depends on patient characteristics and surgical factors. OBJECTIVE: To explore the surgical and patient-related factors that may influence VO2 during the early postoperative period. DESIGN: Prospective study. SETTING: Postoperative intensive care unit. PATIENTS: Study participants were 50 consecutive patients undergoing elective open heart surgery. There were 39 men and 11 women, with a mean age of 58+/-10 years. MEASUREMENTS AND MAIN RESULTS: VO2, oxygen extraction and arterial lactate were measured 1, 4, 12 and 24 h postoperatively. VO2 increased significantly during the first 12 h and stabilized thereafter. Oxygen extraction remained stable through the first 24 h. Covariance analysis on repeated measures showed that the extracorporeal circulatory period (P<0.01), age (P<0.01), body temperature (P<0.05) and use of noradrenalin (P<0.05) were predictive factors influencing postoperative VO2. Although arterial lactate increased significantly during the first 12 h period, no correlation with VO2 was found. However, covariance analysis showed that female sex, patient age (older than 65 years) and bypass period were positive correlating factors for the increase in arterial lactate. CONCLUSIONS: Patient VO2 need is decreased early after open heart surgery and returns to normal after 12 h. Surgical and patient-specific factors are responsible for these changes. Arterial lactate measurements were not found to be reliable indexes of VO2 need during this period. PMID- 10787462 TI - Saphenous vein graft disease treated with the Wiktor Hepamed stent: procedural outcome, in-hospital complications and six-month angiographic follow-up. AB - OBJECTIVES: To evaluate the effectiveness of electively placed heparin-coated stents in the treatment of coronary saphenous vein bypass grafts with de novo lesions less than 15 mm in diameter in a prospective study with all eligible consecutive patients presenting to Middelheim Hospital, Antwerp, Belgium between September 1997 and August 1998. PATIENTS AND METHODS: Fifty patients with 53 lesions were studied. Anginal class, risk factors, quantitative coronary angiographic measurements pre- and postprocedure, procedural outcome, in-hospital events, clinical status on discharge, and six-month clinical and angiographic follow-up (in 48 patients) were recorded. All patients received acetylsalicylic acid and ticlopidine, unless known intolerance was present. RESULTS: On average, 1.1 stents/patient were placed in very old saphenous vein grafts (11. 7+/-3.9 years). Procedural success was 98%. Only two non-Q wave myocardial infarctions (MIs) occurred, with no Q-wave MIs and no deaths during hospital stay. Length of hospital stay was short (2. 4+/-1.7 days), and 96% of patients were free of angina on discharge. At six-months' follow-up, two patients had died, one of whom died of a noncardiac cause. One patient suffered a non-Q wave MI. At six months, 86% of patients were free from angina. Minimal luminal diameter decreased from 1.14 mm before to 3.33 mm after stenting and to 2.52 mm at six months. Restenosis was present in 22% of patients (21.6% of lesions). CONCLUSIONS: In a selected population with coronary saphenous vein bypass graft disease, Wiktor heparin coated stents can be delivered with an excellent periprocedural outcome. Six month outcome appears favourable with a low recurrence of angina (18%) and a low rate of angiographic restenosis (21.6%). PMID- 10787463 TI - Atrial fibrillation and the use of warfarin in patients admitted to an acute stroke unit. AB - OBJECTIVES: To examine the use of warfarin in patients with atrial fibrillation (AF) admitted to hospital because of stroke or transient ischemic attack; and to describe the outcome of AF-associated stroke. DESIGN: Review of the medical records of patients, identified from a prospective registry, admitted from January 1, 1994 through December 31, 1996. SETTING: Tertiary care teaching hospital. RESULTS: AF was present in 92 of 722 (13%) patients at the time of admission. Only eight of 60 (13%) patients with ischemic stroke who were known to be in AF before their stroke were taking warfarin. The in-hospital case-fatality ratio for AF patients was more than double that of patients in sinus rhythm (21% versus 9%, respectively, P=0.001). AF patients were less likely to be discharged home (31% versus 59%, P=0.005). Of the 68 AF patients who survived, 74% left hospital taking warfarin. No warfarin-treated patient experienced intracranial bleeding while in hospital or during follow-up. CONCLUSIONS: Patients with AF had more severe strokes than patients in sinus rhythm. A small proportion of patients with known AF were taking warfarin at the time of hospitalization. Bleeding complications were infrequent. Broader implementation of guidelines for the management of AF is justified to reduce the frequency of stroke in this group of patients. PMID- 10787464 TI - Clinical outcome after isolated tricuspid valve replacement: 20-year experience. AB - OBJECTIVE: To examine the early and late results of isolated tricuspid valve replacement (TVR). DESIGN AND SETTING: All isolated TVRs performed at the Montreal Heart Institute, Montreal, Quebec between January 1978 and January 1998 were retrospectively reviewed. Follow-up data on patients were obtained through the valve clinic. PARTICIPANTS: From a total of 79 TVR and 375 tricuspid annuloplasties performed during the study period, 29 patients who underwent 32 isolated TVRs (six mechanical valves and 26 bioprostheses) were included. Patient age ranged from 25 to 70 years (mean 48), and 62% were female. Twenty-seven patients (84%) were in New York Heart Association (NYHA) functional classes III and IV. Previous valve surgery had been performed in 22 patients (69%) among whom nine had undergone TVR. RESULTS: Postoperatively, a permanent pacemaker was implanted in nine patients (28%), and reoperation because of bleeding was required in two patients. Mean follow-up was 67.7 months (93% complete). Serial echocardiography showed prosthesis dysfunction in three patients, requiring two valve re-replacements at 12.8 and 7.7 years after initial surgery. All patients, except three, showed an improvement of their NYHA class. Six patients (19%) died in hospital and seven patients died during late follow-up at a mean of 38.1 months after surgery, including one valve-related death (mechanical valve thrombosis). The actuarial survival rate of all patients was 63% after five years and 47% after 10 years. CONCLUSION: Isolated TVR remains a high risk procedure. Most survivors, however, should expect a better quality of life by the improvement in their NYHA class. PMID- 10787465 TI - Therapeutic potential of platelet-activating factor antagonism in the management of myocardial infarction. AB - BACKGROUND: Antagonists of platelet-activating factor (PAF) reduce myocardial postischemia reperfusion injury when given before the onset of ischemia. However, the effects of PAF antagonists when administered at a clinically modelled time (during ischemia but before reperfusion) are controversial. Moreover, the extended survival (eight day) and the characteristics of scar formation after treatment with PAF antagonists have not been investigated. OBJECTIVES: To determine the therapeutic potential of PAF antagonist TCV-309 for the treatment of regional myocardial ischemia-reperfusion injury; and to determine the effects of TCV-309 on cardiovascular recovery, evolution of scar formation and survival eight days after a myocardial infarction treated with reperfusion. ANIMALS AND METHODS: Swine underwent regional myocardial ischemia for 60 mins by ligation of the left anterior descending coronary artery, followed by reperfusion for eight days. The treated group (n=7) received PAF antagonist TCV-309 (0.1 mg/kg) 45 mins after ligation; the untreated group (n=7) received vehicle only. RESULTS: Untreated animals experienced significantly (P<0.001) lower systemic arterial blood pressure during the reperfusion period than animals treated with TCV-309. Furthermore, untreated animals required significantly more (P<0.01) antiarrhythmic and inotropic support. Only two of seven animals in the untreated group survived, which was significantly different (P<0.05) from the six of seven treated animals that survived for eight days. Morphometric analyses did not show differences between groups in the characteristics of scar formation following reperfusion for eight days. CONCLUSIONS: PAF antagonist TCV-309 improves survival and reduces cardiovascular dysfunctions associated with regional myocardial ischemia reperfusion injury when administered at a clinically modelled time. PMID- 10787466 TI - Adverse effects of corticosteroids on the cardiovascular system. AB - OBJECTIVE: To review the potential adverse effects of glucocorticoid therapy on the cardiovascular system and to provide insight into the mechanisms of these effects. DATA SOURCES: Case reports and studies demonstrating adverse effects of glucocorticoid therapy on the cardiovascular system were examined from a MEDLINE search. Animal data and in vitro studies were identified to provide insight on the mechanisms of these effects. DATA SYNTHESIS: Undesirable effects identified were dyslipidemia, hypertension and left ventricular free wall rupture after myocardial infarction. Elevations of total plasma cholesterol, triglycerides, low density lipoprotein cholesterol and high density lipoprotein cholesterol are often reported. The elevation of various lipid subfractions is likely mediated by increased plasma insulin levels, impaired lipid catabolism and increased lipid production in the liver. Hypertension was shown to be more prevalent in patients treated with high doses of glucocorticoid. The mechanisms are complex, but final pathways include increased systemic vascular resistance, increased extracellular volume and increased cardiac contractility. Glucocorticoids were demonstrated to increase the incidence of left ventricular free wall rupture by delaying myocardial scar formation in the postmyocardial infarction period. CONCLUSIONS: The major adverse effects of glucocorticoids on the cardiovascular system include dyslipidemia and hypertension. These effects may predispose treated patients to coronary artery disease if high doses and prolonged courses are used. Accordingly, corticosteroids should be employed judiciously in patients with other risk factors for cardiovascular disease, and attention should be paid to risk modification. Low dose and alternate day therapy may reduce the incidence of complications in corticosteroid therapy. The mechanisms of these adverse effects are complex and have not yet been fully explained. PMID- 10787467 TI - Polyarteritis nodosa and myocardial infarction. AB - A 35-year-old man with a history of polyarteritis nodosa is presented. The patient presented with acute anterolateral myocardial infarction that was complicated by diffuse coronary artery aneurysms found during cardiac catheterization. The complication of acute myocardial infarction in a patient with diffuse coronary artery aneurysms associated with polyarteritis nodosa is rare. PMID- 10787468 TI - Myocarditis and pericarditis with tamponade associated with disseminated tuberculosis. AB - Tuberculous involvement of the myocardium is relatively rare. Tuberculous pericarditis with tamponade and myocarditis in a young woman with no evidence of immunosuppression and disseminated tuberculosis is described. Three distinct forms of myocardial involvement are recognized: nodular tubercles (tuberculomas) of the myocardium; miliary tubercles of the myocardium; and an uncommon diffuse infiltrative type. The myocardium is involved by a hematogenous route, by lymphatic spread or contiguously from the pericardium. The diagnosis can be made by endomyocardial biopsy if clinical suspicion is strong and echocardiographic findings are suggestive. Antituberculosis drugs may be curative. With an increasing prevalence of tuberculosis, the possibility of potentially lethal myocardial tuberculosis is important to consider. PMID- 10787469 TI - Medicine and the media. PMID- 10787470 TI - Management of patient grievances based on HCFA and Joint Commission rules. AB - It has long been recognized that the patient loses "control" in the hospital setting as we as health care providers become the caregivers. Through informed consent, advance directives, and patient involvement with care management planning, we have attempted to return some of that control back to the patient. Grievance resolution is simply another way to give back to the patient some of the control that may have been lost in the complexity of health care institutions. Patients and their families deserve a process that adequately addresses their concerns about their hospitalization, and HCFA and the Joint Commission have now addressed the specifics of how that process should occur. Although the process appears cumbersome, time-consuming, and costly, I believe it allows for some interpretive leeway for hospitals to develop their own policies and procedures to comply with the COPs. PMID- 10787471 TI - Procurement and use of placental cord blood. 2: An ethical and legal overview. PMID- 10787472 TI - Stopping embryo research. PMID- 10787473 TI - Stopping science: the case of cryptography. PMID- 10787474 TI - Is self-regulation enough today?: Evaluating the recombinant DNA controversy. PMID- 10787475 TI - Stopping science: human cloning--should it be stopped? PMID- 10787476 TI - Ignorance is not bliss: why a ban on human cloning is unacceptable. PMID- 10787477 TI - Beyond the Pap. What you should know about a new gene test that could someday replace the Pap smear. PMID- 10787478 TI - Hospital system tries both fast and far-reaching strategies to reduce errors. AB - A multihospital system in Boston is doing its best to codify the procedures and and invest in the infrastructure that will minimize the risk of medication errors. PMID- 10787479 TI - Asthma program links hospital resources to physicians, schools. AB - Maine hospitals are working to develop the tools and systems that will better track and treat children with asthma, both in the hospital and in the community. PMID- 10787480 TI - Employee satisfaction starts with communication, management. AB - The bells and whistles of better benefits and other perks may please employees, but they're no substitute for the essentials that make day-to-day work rewarding. To motivate staff to perform at their best, healthcare organizations are investing in front-line managers' skills and backing up their efforts with solid two-way communication. PMID- 10787481 TI - Through compacts, health systems and physicians reshape assumptions. AB - Changing times in healthcare require a rethinking of the premises on which healthcare organizations and physicians have based their relationships. A new compact may call for interdependence, collaboration and other changes that undermine physicians' cherished autonomy, but it may also prove a vital steering mechanism for an organization's future. PMID- 10787482 TI - Shrinking coverage for retired workers. PMID- 10787483 TI - Healthy minds have great ideas. PMID- 10787484 TI - The double edge of a Medicare drug benefit. PMID- 10787485 TI - Applying lessons from the work of art. PMID- 10787486 TI - Target your health dollars to the root causes. PMID- 10787487 TI - The annual managed care survey. Can employers halt the price hikes? PMID- 10787488 TI - Managing care in rural America. PMID- 10787489 TI - This is not your father's EAP. PMID- 10787490 TI - Centrifugal forces unsettle provider networks in Ohio. PMID- 10787491 TI - Data watch. Coordinating the attack on disability costs. PMID- 10787492 TI - Medical privacy regs hit employers on many fronts. PMID- 10787493 TI - Rethinking public health: new training for new times. AB - The new public health agenda will require major changes in the way health authorities, local authorities, Trusts and Primary Care Groups organise and manage their activities. The requirement is for inter-agency co-ordination and inter-professional and inter-sectoral working to a shared agenda, yet the human and resources development planning to achieve these goals has not been done. This paper summarises the key training issues and argues for a collaborative, decentralised and quality assured approach to multidisciplinary public health management education and training. Only with such a joined up human resources plan can Our Healthier Nation succeed where The Health of the Nation signally failed. PMID- 10787494 TI - Taking the lead in fundholding. AB - The prevalence of rather negative, or at best mixed, attitudes of doctors towards participation in management has been reported in several previous research studies. This paper adds to that growing empirical literature, but complements most of the previous studies by focusing on general medical practitioners in primary care rather than on doctors in secondary care settings. The focus for the study were doctors who had agreed to undertake the lead partner role in the now defunct general practitioner fundholding initiative within the UK National Health Service. The findings indicate widespread reluctance but the presence of a minority of enthusiasts, which resonates with previous studies of secondary care settings. A finding of potential significance for future research and practice was that some doctors who were initially reluctant became more keen on undertaking a management role once they had experienced it. PMID- 10787495 TI - An exploratory study of bed management. AB - This paper analyses the role of bed managers and the processes involved in admission, stay, transfer and discharge of patients in the hospital setting. The paper seeks to begin a discussion of the difficulties entailed in the allocation of beds within the context of confined resources. This is achieved by: a review of the somewhat sparse literature on bed management and associated issues; the development of frameworks of analysis with regard to what bed managers do and the information used to support the bed management function; and an explication of results from fieldwork. This is followed by a discussion of the scope of responsibility and career role of the bed manager as well as the potential and problems of bed data. Contacts with others investigating this field and other trusts indicate that the situation in Greater Manchester may be typical of most areas. PMID- 10787496 TI - Microcomputer-based workforce scheduling for hospital porters. AB - This paper focuses on labour scheduling for hospital porters who are the major workforce providing routine cleansing of wards, transportation and messenger services. Generating an equitable monthly roster for porters while meeting the daily minimum demand is a tedious task scheduled manually by a supervisor. In considering a variety of constraints and goals, a manual schedule was usually produced in seven to ten days. To be in line with the strategic goal of scientific management of an acute care regional hospital in Hong Kong, a microcomputer-based algorithm was developed to schedule the monthly roster. The algorithm, coded in Digital Visual Fortran 5.0 Professional, could generate a monthly roster in seconds. Implementation has been carried out since September 1998 and the results proved to be useful to hospital administrators and porters. This paper discusses both the technical and human issues involved during the computerization process. PMID- 10787497 TI - Organizational barriers and ethnicity in the Scottish NHS. AB - The NHS faces a crisis in terms of staffing and recruitment. Many of the ethnic minority GPs in inner city areas throughout the UK are coming up to retirement age, and there is an insufficient supply of trainees to fill estimated vacancies. Over 2,000 nursing vacancies exist across the UK, and recruitment to the profession and retention within the profession are poor. Nurses have been recruited from overseas for the past 40 years, and are currently being recruited from Finland, Malaysia, and the West Indies, whilst doctors are being sought in India, Pakistan and Africa. Overseas recruitment is not a new phenomenon, and numerous studies have been carried out to examine equal opportunities and racial discrimination within the NHS. The aim of this paper was to examine ethnicity and equal opportunities within the Scottish NHS and record the levels of organisational awareness of ethnicity and equal opportunities' issues. The paper also examines the link between health service delivery to ethnic minorities and internal cultural attitudes to staff. PMID- 10787498 TI - Implementing change in the National Health Service. AB - This paper will outline the current changes being imposed on the National Health Service. The literature on change management will be employed to propose some guidelines for health service managers. The National Health Service (NHS) spent much of the 1980s and 1990s learning about the transition from administration to management and must now make the transition from management to leadership. The emphasis is now focused less on doing and more on being. PMID- 10787499 TI - Best practice organisational effectiveness in NHS Trusts. Allington NHS Trust case study. AB - Measuring organisational effectiveness in a health care delivery context is a challenging task. Although there are numerous performance assessment models, audit tools and managerial diagnostic tools, they all tend to fall short in their attempts to closely scrutinize how health care organisations deploy their capabilities to deliver optimum quality in service provision and what performance levels they achieve as a result of their approach. The project reported here attempted to address the aforementioned issues. It set out to examine health care providers from an integrated perspective and to assess the key drivers of effective organisational performance using an adapted model based on the 7s' principles. The research project culminated into a major report highlighting best practices found to be inherent in 18 well performing health care providers. This paper reflects the experience of one of a series of Trusts whose approach to organisational effectiveness was closely examined. PMID- 10787500 TI - The introduction of patient process re-engineering in the Peterborough Hospitals NHS Trust. AB - Patient process recognition and re-engineering (PPR) has become a major concern of recent health care development and management. This paper discusses the position of the National Health Service (NHS) in the UK; where it is at present and where it aims to be. It suggests that the work of the current government in developing community care is central to the work of both the Leicester Royal Infirmary and the Peterborough Hospitals NHS Trust, when building relationships between primary (community) and secondary (hospital) health care provision. This paper aims to examine whether and how PPR can improve patient processes in the NHS. It does this through a case study of PPR in Peterborough Hospital. PMID- 10787501 TI - Outsourcing to increase service capacity in a New Zealand hospital. AB - Service firms manage variability using both demand-side tactics (levelling customer demand), and supply-side tactics (increasing available capacity). One popular way of increasing available capacity is the outsourcing of non-core services. This article uses a case study to examine the impact of an outsourced non-core service on a hospital's overall service system. Findings show that the outsourced service provides access to more sophisticated technology, increases in house capacity and saves capital expenditure. However, the outsourcing also increases the scheduling problems that the hospital faces. These problems are largely due to communication delays from the involvement of more than one organisation. These delays decrease the response time available to match changes in demand for the outsourced service. Given the obvious benefits of such outsourcing, the article concludes that management should pay close attention to the communication pathways between organisations, in order to minimise the end effects identified in this study. PMID- 10787502 TI - Thinking in the old way and trying to live in the new. AB - Professor Viatcheslav Alexeev offers freelance journalist Sarah Powell his views of the state of the Russian national health system and the challenges faced as Russia moves from a socialist to a capitalist economy. Viatcheslav Alexeev is Professor of International Health and Management at the Department of International Health of the Russian Medical Academy of Advanced Medical Studies in Moscow. His wide-ranging experience includes teaching management and health administration, health manpower development, and interpersonal relationships at the Central Institute for Advanced Medical Studies. Following this, for over 20 years he was Assistant Professor and Director of the WHO International Courses for Health Administrators. From 1981 to 1987, he worked on the staff of the Health Manpower Development Division of the WHO in Geneva, in which capacity he gained extensive experience of training, research and organization in over 30 countries. PMID- 10787503 TI - After the internal market: public ownership or poisoned chalice? AB - This article examines the reasons why the internal market failed to gain public acceptance and looks beyond the popular view which argued the non-acceptance was a result of the "two tier" service and increased bureaucracy. It compares the experience of customers in the internal market with those of other public services that have undergone privatization. The article concludes that differences in the range and pattern of the services provided by the NHS led the public to a lower level of understanding regarding improvements and efficiency. The benefits of the market in other once public services were more transparent and there were greater incentives to accept the change. The article recommends that the clinicians involved in the management of primary care groups should learn the lessons of the internal market because users are ill-equipped for the next stage of healthcare reform as they were for the last. PMID- 10787504 TI - Designing an ambulatory clinical practice for outcomes improvement: from vision to reality--the Spine Center at Dartmouth-Hitchcock, year one. AB - Development of a new program for diagnosis and treatment of spine-related problems provided a unique opportunity to design and implement a new model for delivery of health care incorporating outcomes measurement and improvement. Key features include: application of microsystem thinking and interdisciplinary practice; integration of a uniform outcomes measurement tool, the Dartmouth Clinical Value Compass; and touch pad technology for data collection. This, for the first time, provided clinically meaningful point-of-service data and aggregated information for improvement. A further advantage was the ability to integrate a clinical research program within this microsystem. A multisite clinical research trial, the Spine Patient Outcomes Research Trial (SPORT), modeled on the Spine Center microsystem and funded by The National Institute of Arthritis, Musculoskeletal and Skin Diseases and the Office of Research on Woman's Health, the National Institutes of Health, and the National Institute of Occupational Safety and Health, the Centers for Disease Control and Prevention, is currently underway. The significant problems we face today cannot be solved by the same level of thinking that created them. PMID- 10787505 TI - Managed care regulations: impact on quality? AB - Over the last decade, the general public and providers have become increasingly concerned about the quality of care provided by managed care organizations. This article focuses on state and federal legislative proposals to protect quality, and the potential tradeoffs involved with increased regulatory oversight of managed care proposals. In enacting managed care legislation, policy makers should balance the costs potential with the extent to which proposed policies will protect consumers from harm, enhance the operation of the market, or affect quality of care, access to services or choice of providers. PMID- 10787506 TI - Improving HEDIS measurement: linking managed care organization and health system ambulatory care data. AB - This article describes one health system's efforts to improve HEDIS measurement by integrating claims information from its managed care organization with data from its medical center's automated billing, scheduling, and clinical information systems. The authors discuss problems encountered while establishing an integrated measurement process and offer suggestions for others considering such an approach. PMID- 10787507 TI - Content analysis of critical incidents to redesign a university walk-in clinic. AB - The UCLA walk-in clinic was scheduled to move into a new building in late 1997. This prompted the reevaluation of its existing service delivery system. A study was conducted among patients to understand their expectations and perceptions of the clinic's performance. The study was successful in identifying trends and problems and it was instrumental in developing proposals for a new patient flow system to be implemented in the new facility. PMID- 10787508 TI - Communicating outpatient perception to improve quality management. AB - The authors report a study of outpatients' perceptions of clinic waiting times in relation to their expectations and to waiting time norms. The waiting time norm for online registration was 10 minutes, and for blood sampling was 20 minutes. Satisfaction with the behavior of clinic nurses also is reported and analyzed. PMID- 10787509 TI - Quality of life as an outcome-based evaluation of coronary artery bypass graft critical paths using the SF-36. AB - The Medical Outcomes Study Short Form 36 (SF-36) may prove to be a useful tool to evaluate the impact of critical paths (path) on discharged patients. Its ability to measure effects of the path must first be evaluated. The purposes of this study are to evaluate the path using the SF-36, and evaluate its ability to measure path outcomes. Subjects cared for through a coronary artery bypass critical path completed the SF-36 and were compared against a similar baseline sample. The specificity to measure the path influence was inconclusive, because comorbidities and activity restrictions present misleading results in the measurement of disability in its current use. PMID- 10787510 TI - Ambulatory care in the new millennium: the role of consumer information. AB - The past decade has been marked by extensive change in the organization of health care delivery systems (the "supply side" of health care). There has been relatively less change in the nature of the health care consumer (the "demand side" of health care). The emergence of the Internet as a consumer health care technology, however, should significantly affect the nature of demand. The Internet will prepare health care consumers to better express and evaluate care against their preferences. This will create additional pressure on--but significant positive opportunity for--ambulatory care and ambulatory care providers. PMID- 10787512 TI - At the millennium, how happy are CIOs? PMID- 10787511 TI - Interview with Michael R. Callahan, JD. PMID- 10787513 TI - Neuro-biological applications of information technology. PMID- 10787514 TI - Document imaging and workflow technology saves $1.2 million annually. PMID- 10787515 TI - Internet-healthcare: ultimate savior of the system? PMID- 10787516 TI - Q&A: integrated networks. PMID- 10787517 TI - Unique partnership creates multi-functional EDI solution. PMID- 10787518 TI - Healthy dose of data helps California hospital save millions. PMID- 10787519 TI - Timely information is key to critical care. PMID- 10787520 TI - IT helps manage patients with chronic illness. PMID- 10787521 TI - Caring about healthcare. PMID- 10787522 TI - Healthcare 2020: technology in the new millennium. PMID- 10787523 TI - Imaging automation system cuts processing time 83%. PMID- 10787524 TI - Telehealth rescues isolated patients. PMID- 10787525 TI - Hotlist. Document imaging and management. PMID- 10787526 TI - Closing the loop on medication errors. PMID- 10787527 TI - Has consultants' spigot slowed as providers ponder waste? PMID- 10787528 TI - The new machine--move over data! PMID- 10787529 TI - Achieving a healthy e-business solution. PMID- 10787530 TI - A look at healthcare IT from the financial suite. Panel discussion. PMID- 10787531 TI - Charting a new course. PMID- 10787532 TI - After Y2K: it's back to the future. PMID- 10787533 TI - "Patient-doctor digital divide"--online but uncommunicative? PMID- 10787534 TI - Best of the best: IT solutions of 1999. PMID- 10787535 TI - Electronic clinical image management lowers LOS. PMID- 10787536 TI - Courting disaster. PMID- 10787537 TI - Hotlist. Consultants. PMID- 10787538 TI - Harnessing technology for better patient care. PMID- 10787539 TI - Predictive modeling points way to future risk status. PMID- 10787540 TI - Index predicts individual service use. PMID- 10787541 TI - Internet solution reduces medical errors. PMID- 10787542 TI - The promise of converged networks. PMID- 10787543 TI - Convergence in the post-PC era. PMID- 10787544 TI - Put IT in the picture before the merger takes place. PMID- 10787545 TI - The truth about managed care decisions. PMID- 10787546 TI - Software helps doctors get background on patients. PMID- 10787547 TI - Materials management: cutting time and expense. PMID- 10787548 TI - Using the Internet to improve daily operations. PMID- 10787549 TI - Clinical decision support via the Internet. PMID- 10787550 TI - What works. PACS perfect: bridging the image and information gap. PMID- 10787552 TI - Call center provides service with a voice. PMID- 10787551 TI - What works. Managing merger mayhem. PMID- 10787553 TI - Biometrics, technology and Nostradamus.... PMID- 10787554 TI - Hotlist. Decision support. PMID- 10787555 TI - Maximizing the online education experience. PMID- 10787556 TI - Testing the cyber-couch. PMID- 10787557 TI - A question of standards. PMID- 10787558 TI - Partners in accident prevention. PMID- 10787559 TI - Technology can meet field's tough demands. PMID- 10787560 TI - Moving toward collaboration. PMID- 10787561 TI - The Web changes everything. PMID- 10787562 TI - From information to a system. PMID- 10787563 TI - A failure to communicate. PMID- 10787564 TI - Are there standards yet? PMID- 10787565 TI - ACT is growing for a reason: it works. PMID- 10787566 TI - 'Post-merger stress syndrome'. PMID- 10787567 TI - Business plan 101. PMID- 10787568 TI - Hospital pharmacy data: hospital activity. PMID- 10787569 TI - The impact of managed care on pharmacy practice. AB - As prescription drug costs continue their rapid rise, managed care organizations (MCOs) are giving greater attention to containing drug costs. This article examines the impact of managed care on pharmacy practice and reviews types of reimbursement for managed care prescriptions. The author outlines cost containment measures for MCOs and pharmacy benefit management companies, including the use of mail order pharmacies, electronic claims payment, establishment of formularies, therapeutic interchange and generic incentives, and other measures. Cost-containment steps for pharmacies focus on identifying dispensing costs, decreasing overhead, and reducing ingredient costs. The author recommends that pharmacists begin to demonstrate and establish the value of their cognitive services, especially in patient counseling, in order to obtain reimbursement for these services. PMID- 10787570 TI - Pilot study proves CM's effectiveness. PMID- 10787571 TI - Physician/CM team boosts diabetes documentation. PMID- 10787572 TI - New survey measures diabetes satisfaction. PMID- 10787573 TI - Diabetic ketoacidosis path to automate standing orders. PMID- 10787574 TI - How to measure the value of case management. PMID- 10787575 TI - Intensity modulated radiation therapy: new hope for winning the war against cancer. PMID- 10787576 TI - New strategies in opioid therapy for cancer pain. AB - BACKGROUND: The last decade has seen a great increase in the range of opioid agents and formulations available for the treatment of chronic pain, raising new clinical questions and options. METHODS: The author conducted an extensive review of the existing medical literature on opioid agents and formulations currently available. RESULTS: Opioids remain the cornerstone of pharmacotherapy for cancer pain. In addition to severity of pain, coexisting disease, response to previous therapy, the drug's pharmacokinetics and available formulations influence the choice of an opioid agent. Short-half-life drugs, such as morphine, hydromorphone, fentanyl, oxycodone, and oxymorphone, are generally favored initially because they are easier to titrate than long-half-life agents. Long acting controlled-release opioid preparations can lessen the inconvenience of around-the-clock administration of drugs with a short duration of action. In recent years several new formulations have been developed, including controlled release morphine suppositories and suspensions; controlled-release tablets of oxycodone, hydromorphone, and codeine; and transdermal fentanyl, a patch that allows 3-day dosing and avoids the first-pass effect of the liver. Most patients who receive controlled-release opioids should be provided with a rescue dose of an immediate-release opioid to treat pain that may break through the controlled release schedule. CONCLUSIONS: Among the important advances in the last decade in the treatment of chronic pain is the realization that the goals of care and the best means of achieving them differ not only among individuals but often for a single patient throughout a long course of care. Physicians must not only be knowledgeable but also flexible in their approach to managing cancer pain. An understanding of the range of opioid agents and the formulations available can allow physicians to maintain the best possible quality of life for their patients with chronic pain. PMID- 10787577 TI - Marketplace. Universities are telling their hospitals: goodbye, we don't want you any more. PMID- 10787578 TI - Perspectives. Two reports help reset agenda for resumption of Medicare reform debate. PMID- 10787579 TI - Perspectives. Explaining local variations in private coverage rates: it's the labor market. PMID- 10787580 TI - Public reporting of performance: lessons from the USA. PMID- 10787581 TI - Sample size calculations for cluster randomised trials. Changing Professional Practice in Europe Group (EU BIOMED II Concerted Action). AB - OBJECTIVES: Cluster randomised trials, in which groups of individuals are randomised, are increasingly being used in the health field. Adopting a clustered approach has implications for the design of such trials, and sample size calculations need to be inflated to accommodate for the clustering effect. Reliable estimates of intracluster correlation coefficients (ICCs) are required for robust sample size calculations to be made; however, little empirical evidence is available on their likely size, and on factors which influence their magnitude. The aim of this study was to generate empirical estimates of ICCs and to explore factors which may affect their magnitude. METHODS: Empirical estimates of ICCs were calculated for both process variables and patient outcomes from a number of datasets of primary and secondary care implementation studies. RESULTS: Estimates of ICCs varied according to setting and type of outcome. Estimates of ICCs for process variables were higher than those for patient outcomes, and estimates derived from secondary care were higher than those from primary care. ICCs for process variables in primary care were of the order of 0.05-0.15, whilst those in secondary care were of the order of 0.3. Estimates for patient outcomes in primary care were generally lower than 0.05. CONCLUSIONS: Adopting cluster randomisation has implications for the design, size and analysis of clinical trials. This study gives an insight into the potential size of ICCs in primary and secondary care, and provides a practical guide to researchers to aid the planning of future studies in this area. PMID- 10787582 TI - Effect of age on health state valuations. AB - OBJECTIVES: Valuations for health states defined by the EQ-5D measure of health related quality of life, which were elicited using the time trade-off method, from a large sample of the UK general population have previously been shown to be related to the age of the respondent. This paper presents population valuation tariffs for all EQ-5D health states for those aged 18-59 and those aged 60 and over. METHODS: Health state valuations were elicited from a representative sample of over 3000 members of the UK general population using one-to-one interviews. The same regression technique which had previously been used to estimate the EQ 5D tariff for the whole population is used in the current analysis. Additional variables were created which were the product of the original variables and the dummy attached to the age group of the respondent. An earlier qualitative study had suggested that some of the observed differences in valuations might have resulted from more older respondents doubting the plausibility of the 'worse than dead' scenario. Therefore, the model was estimated both when the scores of those aged 60 and over were unmodified and when they were modified to account for this possibility. RESULTS: The tariff based on values from those aged 60 and over, was considerably lower than that based on values from those aged 18-59. When changes between health states were calculated, the difference could be as much as 0.15 on a scale between -1 and 1. However, when the scores of older respondents were adjusted, on the assumption that their lower values for states rated as worse than dead were due to an experimental artefact, the differences between the valuation tariffs of the two age groups greatly diminished. CONCLUSION: These results suggest that whose values are used may have an important effect on health state valuations, and in turn, therefore, on resource allocation decisions, but more research is needed to understand the source of the differences between the values given by different population subgroups. PMID- 10787583 TI - Does general practitioner gatekeeping curb health care expenditure? AB - OBJECTIVES: It is generally assumed that health care systems in which specialist and hospital care is only accessible after referral by a general practitioner (GP) have lower total health care costs. In this study, the following questions were addressed: do health care systems with GPs acting as gatekeepers to specialized care have lower health care expenditure than those with directly accessible specialist care? Does health care expenditure increase more rapidly in countries without a referral system than in those with the GP acting as a gatekeeper? METHODS: Multiple regression analyses on total and ambulatory health care expenditure in 18 OECD countries. RESULTS: Analyses showed only one statistically significant effect (P < 0.05) in countries with gatekeeping GPs: ambulatory care expenditure has increased more slowly than in non-gatekeeping systems. No significant effects of gatekeeping were found on the level of ambulatory care costs, or on the level or growth of total health care expenditure. As in earlier studies, the most important factor influencing aggregate health care costs and their growth is gross national product (GNP), followed by the share of public funding. Relationships that exist at a micro level (such as lower costs with a gatekeeping primary care doctor) did not show up in aggregate data at a macro level. CONCLUSIONS: Gatekeeping systems appear to be better able to contain ambulatory care expenditure. More research is necessary to understand micro level mechanisms and to distinguish the effects of gatekeeping from other structural aspects of health care systems. PMID- 10787584 TI - In pursuit of patient-centred outcomes: a qualitative evaluation of the 'Measure Yourself Medical Outcome Profile'. AB - OBJECTIVES: To ascertain the ability of a patient-generated outcome measure, the 'Measure Yourself Medical Outcome Profile' (MYMOP), to reflect the outcomes of consultations which patients consider are most important, as derived from qualitative interviews. METHODS: A qualitative study using a constant comparative method to analyse semistructured interviews which were then compared with the results of MYMOP questionnaires. A variable-orientated analysis was used to develop themes and a case-orientated analysis was used to develop narrative summaries called vignettes from the interviews. The MYMOP questionnaire was completed by the interviewees (n = 20), a minimum to twice and a maximum of nine times over a four-month period. The scores were displayed on individual MYMOP charts. For each individual, the outcome as measured by MYMOP was compared with the outcome data from the qualitative interviews. The interviewees were attending a variety of complementary practitioners in primary care. RESULTS: The treatment effects which people described were encompassed by five themes: reduction in symptoms; reduction in disability; reduction in, or avoidance of, medication; gaining control and improving coping skills; and securing support and hope through the patient-practitioner relationship. The vignettes demonstrated how individuals identified and valued these various effects and weighed them up in evaluating the overall benefit of the treatment. Their MYMOP charts were able to measure some effects better than others. CONCLUSIONS: The treatment effects, as quantified using MYMOP, accorded with those described by most patients at interview, but some important limitations were identified with MYMOP, particularly an underplaying of the importance of medication avoidance or medication reduction in this patient group. This dimension is lacking in most outcome questionnaires and has been included in a new version of MYMOP. This study also showed that qualitative interview data can help in evaluating the ability of outcome questionnaires to measure the treatment effects that particular patient groups consider most important. PMID- 10787585 TI - Testing evaluation to the limits: the case of English Health Action Zones. PMID- 10787586 TI - Near-patient tests in primary care: setting the standards for evaluation. AB - Near-patient tests (NPT) are an area of rapid technological advance that may enhance the quality of care given by primary care physicians by enabling more accurate diagnosis or more reliable monitoring of therapy. Unfortunately, many new diagnostic technologies prove disappointing in practice, either because of limitations of the technology or because of a failure to address setting-specific issues that were not apparent during their evaluation. A recent systematic review of NPT in primary care has highlighted the methodological weaknesses of most published evaluations. There is a need for a formal statement of the criteria by which to judge the quality of evaluations in this field. In a similar fashion to Wilson and Junger's criteria for screening, seven questions need to be addressed. These cover the performance, reliability and acceptability of the test, its impact on decision-making, comparison with existing practice, cost-effectiveness of the test and integration with the practitioner's knowledge base. Evaluations of NPTs should therefore address the following methodological issues: demonstration of the need for the NPT, definition of the appropriate reference standard, calculation of sample size based on modelling of the decision problem and defining acceptable limits to the likelihood ratio, avoidance of verification bias, selection bias and treatment paradox, testing for the presence of spectrum bias, reporting the sensitivity of the results to changes in setting or prevalence or disease and calculating cost-effectiveness. PMID- 10787587 TI - Conceptual and practical difficulties with the economic evaluation of health services developments. AB - Five recent economic evaluations comparing hospital at home schemes with acute hospital care faced remarkably similar problems. This paper outlines these problems and considers what strategies can be derived from these experiences, which will be relevant to economic evaluations of other aspects of the organisation of care, particularly those crossing the interfaces between primary and secondary health care or the interface between health and social services. The difficulties experienced can be divided into conceptual and practical problems. Conceptual problems were primarily associated with issues of context and related to the choice of comparator, capacity constraints and size of schemes, and the choice between a short or a long run perspective. Practical problems were connected with the time at which schemes were evaluated, the type of clinical study alongside which studies were conducted and the types of data available for use in the analysis. Strategies which can be pursued in conducting economic evaluations of organisational change include giving greater attention to conceptual and hence contextual problems as well as reporting these contextual issues in detail, accepting the need for repeated economic evaluations as organisational changes become more widespread and considering carefully the clinical study design where economic evaluations of organisational change are conducted alongside. These strategies are of importance not just to those conducting economic evaluations but also to those funding appraisals of changes in the organisation of care. Use of different strategies such as those suggested here should be evaluated. PMID- 10787588 TI - The British quasi-market in health care: a balance sheet of the evidence. AB - OBJECTIVES: To summarise the findings from a comprehensive review of research on the effects of the three main elements of the quasi-market reforms of the UK National Health Service (NHS) introduced in 1991/92: General practices becoming fundholders by volunteering to purchase elective care for their patients; Health authorities becoming purchasers of emergency, unplanned and elective services, together with a range of alternatives to fundholding operating under their auspices; The conversion of providers of hospital and community health services to NHS trusts separate from their local health authorities. METHODS: Published and unpublished studies which included any data on the impact of the three main planks of the quasi-market changes, produced between 1991 and late 1998, were identified using a combination of electronic databases, library catalogues at the King's Fund, London, bibliographies, reference lists of individual studies, a survey of NHS directors of public health and consultations with subject area experts. Each main element of the quasi-market was assessed in relation to its impact on: efficiency (primarily productivity); equity; quality; choice and responsiveness; and accountability. RESULTS: There was relatively little measurable change that could be related unequivocally to the core mechanisms of the quasi-market. CONCLUSIONS: The incentives were generally too weak and the constraints too strong to generate the consequences predicted by either proponents or critics of the quasi-market. On the other hand, the way in which the NHS operates was changed irrevocably by the reforms. PMID- 10787589 TI - Opening Pandora's box? Freedom of information and health services research. AB - The introduction, or extension, of freedom of information legislation in Britain and other Western countries seems certain to influence the climate for health services research. However, researchers should be clear that, although such legislation may improve access to certain types of data, it will also create new problems and dilemmas. Statutory freedom of information regimes are likely to raise difficult issues of adaptation and compliance for health service bodies. Researchers may find that this translates into reluctance to agree access and accentuates difficulties in managing field relationships. They may, inter alia, face difficult dilemmas in deciding whether, or how, to report non-compliance, judging whether to extend research into areas where consent has not been obtained and weighing the benefits of using the official rules to insist on disclosure against the dis-benefits of jeopardising relationships. Health services researchers should regard freedom of information legislation, not simply as a resource, but a topic deserving study in its own right. PMID- 10787590 TI - Death by questionnaire: quality of life measurement could seriously damage your health (Vol 4 No 3 pp 129-130) PMID- 10787591 TI - Why shouldn't people who can afford it pay for their own health care? PMID- 10787592 TI - What is clinical effectiveness? PMID- 10787593 TI - Is there any point in spending money on reducing waiting lists? PMID- 10787594 TI - Where is the boundary between clinical and social need for Viagra? PMID- 10787595 TI - Who answers 'willingness to pay' questions? AB - OBJECTIVES: The objectives of this study were twofold. The first was to compare characteristics of responders and non-responders to a survey of women attending a bone mineral density screening service in Aberdeen concerned with the screening process which contained questions on attenders' willingness to pay (WTP) and willingness to wait (WTW) for screening. The second objective was to compare the characteristics of those responding to either the WTP or the WTW questions relative to those who responded to both. METHODS: After receiving a scan, women completed the questionnaire at the clinic or returned it by post. Logistic regression analysis was used to compare the characteristics of the responders and non-responders. RESULTS: Those who smoked were less likely to return the questionnaire, whilst those who drank alcohol were more likely to return it. The majority of respondents answered both WTP and WTW questions. The proportions responding to the WTW and WTP questions were 93.2% and 81.5% for the two questions, respectively (95% confidence interval of difference = 9.4% to 13.9%). The only result which was statistically significant at the 1% level showed that, relative to those who answered the WTW but not the WTP questions, those who answered both were more likely to be older when they left full-time education. A weaker statistical association (at the 5% level) revealed that those who were older when leaving full-time education were more likely to answer a WTP question than not. CONCLUSIONS: WTP questions seem to be less acceptable to those who leave full-time education earlier. Analysts may need to account for this in future studies. Whether such results can be replicated and reasons for non response should be investigated. PMID- 10787596 TI - Strategic management of an electronic patient record project using the balanced scorecard. PMID- 10787597 TI - Implementing an emergency department information system--how complicated can we make this? PMID- 10787598 TI - Efficient and effective business process redesign: case study on enterprise-wide scheduling/express check-in. PMID- 10787599 TI - Automating clinical documentation in dentistry: case study of a clinical integration model. PMID- 10787600 TI - A primer on pharmacy information systems. PMID- 10787601 TI - Essential characteristics of an electronic prescription writer. PMID- 10787602 TI - A computerized physician order entry system for the promotion of ordering compliance and appropriate test utilization. PMID- 10787603 TI - Serving physician information needs: a model for the transition to an electronic medical record. PMID- 10787604 TI - Uniform data standards for capturing patient medical record information at the point of care. PMID- 10787605 TI - Clinical information systems: paving the way for clinical information management. PMID- 10787606 TI - The 1999 National Caring Awards. Jeannie Jaybush--changing the world one diaper at a time. PMID- 10787607 TI - The 1999 National Caring Awards. Irasema Salcido--reinventing public education. PMID- 10787608 TI - The 1999 National Caring Awards. Peter Samuelson--empowering children and making their dreams come true. PMID- 10787609 TI - The 1999 National Caring Awards. Cathrine Sneed--growing flowers, trees, and people. PMID- 10787610 TI - The 1999 National Caring Awards. Arthur & Mary Willhite--casting bread and love among the homeless. PMID- 10787611 TI - The 1999 National Caring Awards. Lance Secretan--changing the world by reintroducing spirit & value into the workplace. PMID- 10787612 TI - The 1999 National Caring Awards. Young adult awardees--Craig Kielburger, Concord, Ontario. PMID- 10787613 TI - The 1999 National Caring Awards. Young adult awardees--Ashley Anderson, Houston, Texas. PMID- 10787614 TI - Carrying on the Nobel family tradition: combining caring for people and the earth. PMID- 10787615 TI - The 1999 National Caring Awards. Young adult awardees--Nadia Ben-Youssef, Sidney, Montana. PMID- 10787616 TI - The 1999 National Caring Awards. Young adult awardees--Aubyn Burnside, Hickory, North Carolina. PMID- 10787617 TI - The 1999 National Caring Awards. Young adult awardees--Emily Douglas, Powell, Ohio. PMID- 10787618 TI - The 1999 National Caring Awards. Young adult awardees--Jamie Ridgely, Manchester, Maryland. PMID- 10787619 TI - A tribute to John Glenn: American hero. PMID- 10787620 TI - The Frederick Douglass Museum and Hall of Fame for Caring Americans--an intellectual and spiritual oasis on Capitol Hill. PMID- 10787621 TI - The 1999 National Caring Awards. Nancy Brinker--winning the war against breast cancer. PMID- 10787622 TI - Behavioral contract management: a prescription for employee and patient compliance. AB - A health care manager's responsibilities of offering efficient service while ensuring effective outcomes is hampered frequently by both employees and patients whose unacceptable behaviors are detrimental to one or both of these mandates. Behavioral contracts offer a structured method that allows both patients and employees to self-actualize the required behavioral changes. When noncompliant behavior is diagnosed, behavioral contracts are the prescription that places the responsibility for corrective action squarely on the shoulders of the offending individual. PMID- 10787623 TI - What your hospital's CEO is thinking about. AB - The contemporary hospital CEO is concerned about much more than just "big picture" environmental scanning and strategic planning. In fact, he or she is concerned with a host of problems and opportunities whose common denominator involves reconciling contradictions between, for example, mission and margin, risk and return, consensus and momentum, and strategy and opportunism. In this context, the hospital CEO is concerned especially about basic tasks related to focusing the organization, simplifying processes, creating a sense of urgency, and communicating effectively. These objectives often are elusive, yet the CEO ultimately is uniquely accountable for their accomplishment. PMID- 10787624 TI - Will you fit if you move to a job in another culture? AB - In today's health care environment, many health care managers choose to move to a new setting. Because environments vary from setting to setting, a manager who plans to move needs to evaluate potential "fit" in the new locality. An evaluation of "fit" includes an assessment of both the job culture and persons in the environment. The Giger-Davidhizar transcultural assessment model provides a model for evaluating a new job culture. PMID- 10787625 TI - Case in health care management: "the employee who is always right". PMID- 10787626 TI - Striving toward continuous quality improvement: a case study of Saint Mary's Hospital. AB - This case analysis is the result of a year-long study designed to identify and assess the ingredients that led to the successful implementation of a continuous quality improvement (CQI) program at Saint Mary's Hospital in Grand Rapids, Michigan. The key ingredients of success included: (1) an organizational structure and leadership commitment for identifying and improving processes, (2) use of data-based statistical and analytical tools to study processes, (3) empowerment of teams of employees to take charge of the operations of their own work tasks in a manner that encourages continuous learning, (4) involvement of internal and external customers through the improvement process, and (5) development of effective measures for monitoring improvement. The benefits of the CQI efforts at Saint Mary's have been remarkable and hospital-wide. PMID- 10787628 TI - Department Manager's Checklist: absenteeism. PMID- 10787627 TI - The friendly art of persuasion. AB - The ability to persuade is a critical skill for the health care professional in relating to employees, colleagues, and groups of individuals if health care objectives are to be promoted. However, persuasion is an art, which requires careful preplanning and deliberative actions if it is to be carried out successfully. Persuasion done in a friendly manner is more likely to be effective. This article provides techniques that will enable the health care professional to use persuasive methods most advantageously. PMID- 10787629 TI - Managed care evolution--where did it come from and where is it going? AB - The emergence and development of managed care has been and will continue to be a major force in America's health care delivery system. Specifically reviewed in this article are those legislative initiatives that contributed to the formation of this system of reimbursement and utilization management. Also addressed are responsibilities that can be ascribed to all stakeholders in necessitating this system as well as the adjustments required to make the system more responsive to the needs of higher-risk populations. This article concludes with a series of recommended actions that will be required of all individual citizens in assuring the future economic viability of this nation's health care programs. PMID- 10787630 TI - Principles of workforce stability. AB - The low unemployment rate and the shortage of skilled health care personnel stoke the trend for workers to change jobs and employers. To build workforce stability, employers and managers must make special efforts to counter this trend. Workplace stability is achieved by fielding teams of carefully selected workers and taking measures to prevent them from jumping ship. This article relates how employers and managers can accomplish this. PMID- 10787631 TI - Breathing hospital air can make you sick. AB - Indoor air quality has deteriorated so much since the 1970s oil shortage and subsequent energy-efficient construction of buildings that people are becoming seriously ill by just breathing the indoor air. This is a problem with all industrial buildings and hospital staff are at particular risk. There are various things that hospital managers from different departments can do to make the air safe for staff and patients to breathe. PMID- 10787632 TI - Balancing inside and outside: preparing for tomorrow while fulfilling today's responsibilities. AB - Many managers experience a common organizational contradiction in terms of how their attention is focused. One may be primarily internally focused (primarily attentive to people and activities within the department or organization), meaning that it is the job at hand that commands the most attention. Or one may be externally focused (primarily attentive to people and activities outside of the department or organization), meaning that it is the sources of reward and opportunity that command the most attention. Over the long run, the internally focused individual tends to develop more strongly as a manager but the externally focused individual, by virtue of greater visibility, tends to attract more career advancement opportunity. For lasting growth and advancement, the manager requires both solid experience and opportunity; the individual can be neither slave to the daily task nor absentee manager. Rather, a sensible approach to career advancement requires a healthy balance of internal and external focus. PMID- 10787634 TI - Is America ready for terrorism?. Interview by Marie Nordberg. PMID- 10787633 TI - A manager asks: introducing change to the dependent employee and to the group. PMID- 10787635 TI - Documentation in a two-tiered system. PMID- 10787636 TI - Abdominal aortic aneurysm. PMID- 10787637 TI - Special Operations Combat course tests medics' skills. PMID- 10787639 TI - Wanted: skilled EMS personnel. PMID- 10787638 TI - Putting the common back into courtesy. PMID- 10787640 TI - Marketing yourself: designing a resume. PMID- 10787641 TI - Becoming a supervisor: the road to advancement. PMID- 10787642 TI - A world of opportunities--the challenges and benefits of working overseas. PMID- 10787643 TI - EMS incident management: duties of the incident safety officer. PMID- 10787644 TI - When lightning strikes. PMID- 10787645 TI - Gotham City news, Gotham City blues. PMID- 10787646 TI - The evidence base on the cost-effectiveness of malaria control measures in Africa. AB - This review assesses the range and quality of the evidence base on the cost effectiveness of malaria prevention and treatment in sub-Saharan Africa. Fourteen studies are reviewed, covering insecticide-treated nets, residual spraying, chemoprophylaxis for children, chemoprophylaxis or intermittent treatment for pregnant women, a hypothetical vaccine, and changing the first line drug for treatment. The available evidence provides some guidance to decision-makers. However, the potential to inform policy debates is limited by the gross lack of information on the costs and effects of many interventions, the very small number of cost-effectiveness analyses available, the lack of evidence on the costs and effects of packages of measures, and the problems in generalizing or comparing studies that relate to specific settings and use different methodologies and outcome measures. PMID- 10787647 TI - Implementing a malaria curtains project in rural Malawi. AB - This paper describes planning, implementation, monitoring and evaluation activities carried out in support of a malaria control project that used permethrin-impregnated curtains in eight villages in rural Malawi. Findings from formative evaluation and project monitoring aspects of the evaluation are presented. Permethrin-impregnated curtains were introduced to villagers who participated in household self-help projects. To implement the project, village health workers were trained and worked closely with existing project personnel as well as traditional headmen to assure village participation, facilitate health education and coordinate curtain-dipping (impregnation) meetings. A quasi experimental evaluation design used surveys and observations to measure change in cognitive, behavioural and health outcome indicators. Village adoption rates averaged 50%, with variation between villages. Monitoring data showed a high degree of compliance with curtain re-impregnation initially and high perceived efficacy of curtains. Issues discussed include village readiness for change, trust, acceptability of the innovation, cost, sustainability and leadership. PMID- 10787648 TI - Infectious disease policy: towards the production of health. AB - This paper reflects on current infectious disease control methods, their biomedical history and the management structures they are driven by. It asks whether a broader concept of 'infectious disease policy', with an emphasis on 'process' rather than outcomes, will help to re-frame the control discourse to ensure that infectious disease control is not only a method for preventing and treating infectious diseases, but is also a path for the creation of healthy communities. PMID- 10787649 TI - Health for some? The effects of user fees in the Volta Region of Ghana. AB - This paper reports key findings and conclusions from a 1996 study of user fees and exemptions in the Volta Region of Ghana. A variety of data sources and methods were used, including interviews with patients and managers, community based focus group discussions, analysis of facility records and analyses of previous household survey data. Official fee levels and exemption categories were established in 1985. While this legislation made provision for drug fees to be 'at cost' and thus to be revised in line with inflation, other official fees have not been adjusted since 1985. In the face of declining real levels of budget allocations and decreased supplies of essential consumables from the Central medical stores, facility managers have established their own pricing and fee collection systems. This has been allowed by the Ministry of Health, but the decentralized nature of fee setting and collection practices has made it very difficult for the Ministry to monitor the effects of fees. The study found that facility managers have been very active in setting and collecting fees and using the revenues to purchase essential inputs. The level of revenues being mobilized accounts for between two-thirds and four-fifths of the non-salary operating budget of government health facilities, and virtually all of the resources for non-salary operating expenses in mission hospitals. Official exemptions are largely non-functional. Less than one in 1000 patient contacts were granted exemption in 1995. With estimates that between 15 and 30% of the population lives in poverty, the failure of exemptions to function means that fees are preventing access for the poor, or are imposing significant financial hardships on this part of the population. Health facilities in the Volta Region have achieved a kind of 'sustainable inequity', with fees enabling service provision to continue, while concurrently preventing part of the population from using these services. PMID- 10787650 TI - National Health Accounts development: lessons from Thailand. AB - National Health Accounts (NHA) are an important tool to demonstrate how a country's health resources are spent, on what services, and who pays for them. NHA are used by policy-makers for monitoring health expenditure patterns; policy instruments to re-orientate the pattern can then be further introduced. The National Economic and Social Development Board (NESDB) of Thailand produces aggregate health expenditure data but its estimation methods have several limitations. This has led to the research and development of an NHA prototype in 1994, through an agreed definition of health expenditure and methodology, in consultation with peer and other stakeholders. This is an initiative by local researchers without external support, with an emphasis on putting the system into place. It involves two steps: firstly, the flow of funds from ultimate sources of finance to financing agencies; and secondly, the use of funds by financing agencies. Five ultimate sources and 12 financing agencies (seven public and five private) were identified. Use of consumption expenditures was listed under four main categories and 32 sub-categories. Using 1994 figures, we estimated a total health expenditure of 128,305.11 million Baht; 84.07% consumption and 15.93% capital formation. Of total consumption expenditure, 36.14% was spent on purchasing care from public providers, with 32.35% on private providers, 5.93% on administration and 9.65% on all other public health programmes. Public sources of finance were responsible for 48.79% and private 51.21% of the total 1994 health expenditure. Total health expenditure accounted for 3.56% of GDP (consumption expenditure at 3.00% of GDP and capital formation at 0.57% of GDP). The NESDB consumption expenditure estimate in 1994 was 180,516 million Baht or 5.01% of GDP, of which private sources were dominant (82.17%) and public sources played a minor role (17.83%). The discrepancy of consumption expenditure between the two estimates is 2.01% of GDP. There is also a large difference in the public and private proportion of consumption expenses, at 46:54 in NHA and 18:82 in NESDB. Future NHA sustainable development is proposed. Firstly, we need more accurate aggregate and disaggregated data, especially from households, who take the lion's share of total expenditure, based on amended questionnaires in the National Statistical Office Household Socio-Economic Survey. Secondly, partnership building with NESDB and other financing agencies is needed in the further development of the financial information system to suit the biennial NHA report. Thirdly, expenditures need breaking down into ambulatory and inpatient care for monitoring and the proper introduction of policy instruments. We also suggest that in a pluralistic health care system, the breakdown of spending on public and private providers is important. Finally, a sustainable NHA development and utilization of NHA for planning and policy development is the prime objective. International comparisons through collaborative efforts in standardizing definition and methodology will be a useful by-product when developing countries are able to sustain their NHA reports. PMID- 10787651 TI - Feasibility of hospital-based blood banking: a Tanzanian case study. AB - The demand for blood transfusion is high in sub-Saharan Africa because of the high prevalence of anaemia and pregnancy related complications, but the practice is estimated to account for 10% of HIV infections in some regions. The main response to this problem by the international donor community is to establish vertically implemented blood transfusion services producing suitable (safe) blood at a cost of US$25-40 per unit. However, the economic sustainability of such interventions is questionable and it is argued here that hospital-based blood transfusion services operating at a basic adequate level are sufficient for low income African countries. The results of a project aimed at improving such services in Tanzania are presented. The main findings are: (1) the cost per suitable blood unit produced was US$12.4; (2) at an HIV test sensitivity of 93.5% during the study period, discounted financial benefits of the interventions exceeded costs by a factor of between 17.2 and 37.1; (3) the cost per undiscounted year of life saved by use of these interventions was US$2.7-2.8; and (4) safe blood transfusion practices can be assured at an annual cost of US$0.07 per capita. Recommendations are made to ensure safe blood transfusion practices at hospital-based blood banks in Tanzania. PMID- 10787652 TI - Rapid assessment and sample surveys: trade-offs in precision and cost. AB - Health policies in many developing countries are increasingly designed and evaluated by data collected using household surveys. This trend is closely associated with three factors: the increasing complexity of health systems, the increase in pressure for better accountability and improved health system management, and the continuing existence of poorly maintained vital statistics systems. But national household- or individual-level surveys are expensive. These factors have, unsurprisingly, coincided with an increase in the use of rapid survey methods. Although rapid methods have been used frequently, few studies have tested the quality of data obtained by them, and none have compared the cost of the rapid surveys with non-rapid surveys in a developing country context. This paper presents the trade-offs in cost and quality of information obtained from a rapid assessment survey in Ecuador. The results from the rapid survey were compared with results obtained from a national survey conducted six months earlier. The objective was to see what alternative policies might be arrived at if the data from the rapid survey were used in place of the large survey. In addition, the relative costs of obtaining that information were measured. Results indicate that the majority of demographic and contraceptive use measures are similar between the two surveys. The rapid survey was three times as cost efficient as the traditional survey, if relative bias is not taken into account. However, there appears to be selection bias in the choice of households, which distorts the probability of predicting contraceptive use by residence, and makes it appear that rural women are as likely to use contraception as their counterparts in urban Ecuador. This result has implications for the credibility of rapid surveys, and suggests that their use needs to be tailored for particular programmes and policies, and overseen with, at least, a modicum of statistical caution. PMID- 10787653 TI - A comparison of four approaches for measuring clinician time use. AB - Concern about rapid growth in demand for reproductive health services in developing countries has created interest in productivity and costs of existing programmes. Staff costs usually constitute the largest share of total service costs, meriting special effort to ensure that they are measured accurately. Several techniques have been used in the literature to analyze staff activity, but these techniques have not been validated. This paper reports on a study conducted in three Ecuadoran clinics. The study uses an observational time-motion (TM) technique as a benchmark, and compares results from three other techniques to those obtained using TM. None of the alternative techniques produces estimates that agreed with TM estimates; deviations from TM are particularly large for non contact time, defined as clinician activities carried out when clients are not present. Implications of these findings for productivity and cost studies are discussed, and possible avenues for future research are proposed. PMID- 10787654 TI - Successful contracting of prevention services: fighting malnutrition in Senegal and Madagascar. AB - There are very few documented large-scale successes in nutrition in Africa, and virtually no consideration of contracting for preventive services. This paper describes two successful large-scale community nutrition projects in Africa as examples of what can be done in prevention using the contracting approach in rural as well as urban areas. The two case-studies are the Secaline project in Madagascar, and the Community Nutrition Project in Senegal. The article explains what is meant by 'success' in the context of these two projects, how these results were achieved, and how certain bottlenecks were avoided. Both projects are very similar in the type of service they provide, and in combining private administration with public finance. The article illustrates that contracting out is a feasible option to be seriously considered for organizing certain prevention programmes on a large scale. There are strong indications from these projects of success in terms of reducing malnutrition, replicability and scale, and community involvement. When choosing that option, a government can tap available private local human resources through contracting out, rather than delivering those services by the public sector. However, as was done in both projects studied, consideration needs to be given to using a contract management unit for execution and monitoring, which costs 13-17% of the total project's budget. Rigorous assessments of the cost-effectiveness of contracted services are not available, but improved health outcomes, targeting of the poor, and basic cost data suggest that the programmes may well be relatively cost-effective. Although the contracting approach is not presented as the panacea to solve the malnutrition problem faced by Africa, it can certainly provide an alternative in many countries to increase coverage and quality of services. PMID- 10787655 TI - Coverage and cost of iodized oil capsule distribution in Tanzania. AB - Distribution of oral iodized oil capsules (IOC) is an important intervention in areas with iodine deficiency disorders (IDD) and low coverage of iodized salt. The mean reported coverage of 57 IOC distribution campaigns from 1986-1994 of people aged 1-45 years in 27 districts of Tanzania was 64% (range 20-96%). This declined over subsequent distribution rounds. However, due to delayed repeat distribution, only 43% of person-time was covered, based on the programme objective of giving two IOC (total 400 mg iodine) at 2-year intervals. Three different capsule distribution strategies used in 20 distribution rounds in 1992 1993 were analyzed in depth. Withdrawal of financial support for district distribution expenses under the 'district team' strategy, and the subsequent change to integrated 'primary health care' distribution, increased delays and capsule wastage. The third, more vertical strategy, 'national and district teams', accomplished rapid distribution of IOC about to expire and subsequently a return to the initial 'district team' allowance strategy was made. Annual cost of 'district team' distribution was 26 cents per person (400 mg iodine/2 years). Cost analysis revealed that the IOC itself accounts for more than 90% of total costs at the levels of coverage achieved. IOC will be important in the elimination of IDD in target areas of severe iodine deficiency and insufficient use of iodized salt, provided that high coverage can be achieved. Campaign distribution of medication with high item cost and long distribution intervals may be more cost-effectively performed if separated from regular PHC services at their present resource level. However, motivating health workers and community leaders to do adequate social mobilization remains crucial even if logistics are vertically organized. Insufficient support of distribution expenses and health education may lead to overall wastage of resources. PMID- 10787656 TI - Do multi-sectoral development programmes affect health? A Bolivian case study. AB - This cross-sectional study, carried out in Inquisivi, Bolivia, a rural area where Save the Children/US works, tests the hypothesis that participation in multisectoral development programmes results in improved health behaviours and better health outcomes. To test this hypothesis, four groups of households were compared: those participating in Save the Children's health-only programmes; those with access to health and micro-enterprise credit or health and literacy programmes; those participating in all three programmes (health, credit and literacy); and households from comparison communities (no access to any of Save the Children's programmes). Data come from a stratified sample of 499 households in the altiplano, foothills and valleys of the Andes. Findings reported here suggest that there is no clear association between participation in one or more of Save the Children's programmes and parents' actions to prevent and treat diarrhoea. Additionally, the point prevalence of diarrhoea was similar for all four groups. However, children of individuals participating in health, credit and literacy were significantly less likely than children from comparison communities to be malnourished or at risk of becoming malnourished, even after controlling for such potentially confounding factors as social class, source of drinking water, and the availability of health facilities. PMID- 10787657 TI - Evaluating payment mechanisms: how can we measure unnecessary care? AB - There has been substantial concern that linking hospital or physician remuneration to the amount of care provided will encourage excessive provision of health care. Studies that seek to explore this relationship require methods to measure unnecessary care. This paper presents and discusses a method used to assess the magnitude of unnecessary care in the context of an evaluation of the impact of paying bonuses to physicians employed in public hospitals in Shandong Province, China. PMID- 10787658 TI - New focus for community hospitals: neuro centers. PMID- 10787659 TI - Hospitals with residents must improve communication to avoid unionization. PMID- 10787660 TI - Healtheon/WebMD goal: cut $250 billion from U.S.'s $1.3 trillion in health spending. PMID- 10787661 TI - Chicago Heights hospital uses community collaboration to build fitness center. PMID- 10787662 TI - The future of health care. PMID- 10787663 TI - It pays for hospitals to support programs that help control physician stress. PMID- 10787664 TI - Look at some new scenarios and threats. PMID- 10787665 TI - New approaches need to integrate technology. PMID- 10787666 TI - Value of preventive services questionable for HMO patients over 75, study suggests. PMID- 10787667 TI - BBA spurs increase in hospital, post-acute facility mergers, acquisitions. PMID- 10787668 TI - Health care CEOs expect merit increase averaging 4.3% in 2000. PMID- 10787669 TI - Alaska has lowest percentage of Medicare beneficiaries, West Virginia highest. PMID- 10787670 TI - Patients justified in their concern about privacy of health Web sites, study finds. PMID- 10787671 TI - Growing pains: when healthcare facilities merge. AB - The article discusses how the loss prevention services department at a large Midwestern healthcare system was able to meet the security needs of a wide range of disparate affiliates--including a dozen hospitals, clinics, long-term care facilities, pharmacies, a home healthcare service and social service agency, and an array of support facilities. PMID- 10787672 TI - Dealing with school violence: how hospitals met this new challenge to emergency preparedness. AB - This article discusses how hospitals in Jonesboro, AR, and Denver, CO, met the challenge of dealing with school shooting rampages that resulted in multiple casualties and received widespread media coverage. Hospitals need to be well prepared to implement their emergency disaster plans and handle the physical and emotional trauma of such incidents which, according to a well-known criminologist, may become more frequent. PMID- 10787673 TI - Avoiding liability for hospital parking security in the 21st century. PMID- 10787674 TI - NCMEC: clarification of statement on electronic bracelets. PMID- 10787675 TI - Is your hospital prepared to treat hundreds of victims of a terrorist attack? AB - Why and how your healthcare facility should be prepared for an incident of terrorism--nuclear, biological, or chemical. Have a plan, get training, consider protective equipment, and be familiar with the legal issues, the author advises. PMID- 10787676 TI - Managing the threat of terrorism in abortion facilities. AB - Acts of violence by anti-abortion terrorists are a growing threat to healthcare facilities and clinics. This article provides advice on deterring bomb threats, developing a bomb incident plan, ensuring the physical security of a facility, handling a bomb threat phone call or a suspicious object, and dealing with the media. Security plays a vital role in any anti-terrorist plan, says the author. PMID- 10787677 TI - Healthcare security on the front line against domestic violence and stalking affecting the workplace. AB - Domestic violence and stalking affect the health and safety of employees in the workplace. Security and EAP staff, says the author, can give healthcare facilities the edge in front-line deterrence. The article discusses definitions, key factors to consider, and how to go about dealing with these problems. PMID- 10787678 TI - Honesty, the ohana, and individual responsibility. AB - The author discusses how applying the Army values of loyalty, duty, respect, selfless service, honor, integrity, and personal courage has helped his healthcare facility provide excellent care to its patients. PMID- 10787679 TI - Conducting a security audit. AB - Prior to conducting a full-fledged security survey in order to meet requirements of JCAHO and other agencies, it is essential to conduct an initial audit that will assess your hospital's vulnerability. The information from this audit can guide and direct the survey process. This article describes what is involved in preparing for the initial audit and includes a sample security audit report. PMID- 10787680 TI - Security's amazing recovery. AB - A Michigan hospital upgraded its security by forging a partnership with a nearby university. The article describes how the hospital's security presence was transformed from a detached, reactive service into a committed, proactive team while reducing costs for the hospital. PMID- 10787681 TI - Beating down the cost of security system installations and repairs. AB - How a growing healthcare system was able to upgrade its security systems while reducing the costs of installations and repairs and improving quality. The author gives details on what was done, how it was accomplished, and what cost savings have been achieved. PMID- 10787682 TI - Outsourcing: considerations for healthcare protection managers. AB - The author discusses the use of contract services--myths, advantages, and disadvantages. He provides tips on what to look out for and what questions to ask before making a change to contract security. PMID- 10787683 TI - Hurricane Georges and New Orleans hospitals: preparing for a killer storm. AB - This article describes the experiences of six New Orleans healthcare facilities in preparing for and coping with one of the worst potential storms in years. Some of the problems they faced were foreseen. Others were unforeseen and required considerable ingenuity to solve. PMID- 10787684 TI - Your infection control program. AB - Training is essential in order to avoid being infected by a bloodborne disease, says the author. This training must be a cooperative effort by the employer and the safety and/or emergency response employee. The bloodborne pathogens plan must include education, vaccination, and follow-up. PMID- 10787685 TI - Strategic learning in healthcare organizations. AB - There is no definitive blueprint for the healthcare organization involved in strategic learning. However, what distinguishes strategic learning institutions is their acknowledgment that they must discover their own paths and solutions rather than blindly follow a detailed strategic mandate from administration. Answers to their most critical implementation and adaptive questions will not flow down ready-made from above, but will be tailored to meet the requirements of their own particular situation. Strategic learning organizations have certain attributes in common in developing their own answers: They continuously experiment rather than seek final solutions. They favor improvisation over forecasts. They formulate new actions rather than defend past ones. They nurture change rather than permanence. They encourage creative conflict rather than tranquillity. They encourage questioning rather than compliance. They expose contradictions rather than hide them (Weick 1977). Most importantly, strategic learning organizations realize that successful strategic change is best undertaken as a process of learning (O'Sullivan 1999). Healthcare organizations can no longer afford the illusion of traditional strategic planning, with its emphasis on bureaucratic controls from the top to the bottom. They must embrace the fundamental truth that most change occurs through processes of learning that occur in many locations simultaneously throughout the organization. The initial step in discovering ways to improve the capability of healthcare organizations is to adapt continuously while fulfilling their mission. Healthcare leaders must create a shared vision of where an institution is heading rather than what the final destination will be, nurture a spirit of experimentation and discovery rather than close supervision and unbending control, and recognize that plans have to be continuously changed and adjusted. To learn means to face the unknown: to recognize that we do not possess all the answers; to concede that we do not always know what to do; to admit that past actions and solutions may no longer be appropriate, in fact may have been the incubators of today's problems; to question basic assumptions long held about running the institution; and to make ourselves vulnerable to the political dynamics prevalent in all organizations. Hospitals and other healthcare organizations must seek to develop and maintain a continuing state of readiness in which everyone in the organization, from front line clinician to senior management, is poised to act in anticipation of and in response to unforeseen changes in the environment and to learn from their own experiences in confronting the future. PMID- 10787686 TI - An analysis of 1997 healthcare initial public offerings. AB - The corporatization of healthcare is here to stay, and it is influencing much in the industry. Thus, it is important for healthcare executives and professionals to identify and evaluate companies in this sector of the economy. One method is to conduct annual surveys of new healthcare IPOs, obtain data on those companies, and assess growth and decline by following these corporations longitudinally. This study represents the example of establishing a baseline of IPOs in 1997. Since the information is public and relatively easy to access, healthcare professionals can use the wealth of information in a variety of ways. PMID- 10787687 TI - Implementing advance directives: a continuing problem for provider organizations. AB - It has been suggested that widespread use of advance directives might encourage systematic rationing of healthcare, especially to the elderly. If a right to die becomes a duty to die, the living will and its progeny have become a Frankenstein monster. Indeed, public statements by state and federal officials that the elderly should be required to have living wills raised a storm of protest in the past. Regardless of true motives, such suggestions tend to be seen by the public as motivated by economics. The organization must be alert to the issues raised by advance directives. Managers are obliged to obey the law. Beyond that, however, they should work to enhance patient autonomy by facilitating preparation and availability of advance directives and surrogate decision making, should that become necessary. PMID- 10787688 TI - Nursing unions, Part II: Mature organizations lead to shared management. PMID- 10787689 TI - A model of senior care within an integrated healthcare delivery system. PMID- 10787690 TI - Revisiting the roles of need and demand in ensuring program success. PMID- 10787691 TI - Physician-health system partnerships: strategies for finding common ground. PMID- 10787692 TI - Integrating medical group practices that work. PMID- 10787693 TI - Increasing workplace motivation. PMID- 10787694 TI - Using e-mail effectively. PMID- 10787695 TI - Building your leadership character. PMID- 10787696 TI - Organ donation. PMID- 10787697 TI - Can physician unionization succeed? PMID- 10787698 TI - Allocating limited capital resources. PMID- 10787699 TI - Medical errors spur new patient-safety measures. PMID- 10787700 TI - The ins and outs of outsourcing. PMID- 10787701 TI - Interview with ACHE's incoming chairman. Interview by Ellen G. Lanser. PMID- 10787703 TI - Is dropping your association membership safe? PMID- 10787702 TI - Close encounters of the customer service kind. PMID- 10787704 TI - Ethics self-assessment. PMID- 10787705 TI - Would you have thought of anthrax? PMID- 10787706 TI - Fined $100,000--for "dumping" patients he couldn't treat. PMID- 10787707 TI - Doctoring "Down Under". PMID- 10787708 TI - The devastation caused by a decimal point. PMID- 10787709 TI - Invitation to a lawsuit: financial incentives to limit care. PMID- 10787710 TI - Are bargain-basement medications harming your patients? PMID- 10787711 TI - Managed care. States move to tighten their grip on HMOs. PMID- 10787712 TI - Elections 2000. Will patient protection be a pivotal issue? PMID- 10787713 TI - Reimbursement. Are hospitals using physician practices to get larger Medicare payments. PMID- 10787715 TI - Lobbying. Medicare reform resuscitated? PMID- 10787714 TI - Medicare. Seniors still aren't sure what's going on. PMID- 10787716 TI - How does your productivity measure up? PMID- 10787717 TI - Why that computer "bargain" may be anything but. PMID- 10787719 TI - How your colleagues ease the hassle of nursing home care. PMID- 10787718 TI - I finally understand the waiting room ordeal. PMID- 10787720 TI - Should your CEO be an outsider or hired from within? PMID- 10787721 TI - Health care converges: a new era in patient care emerges. PMID- 10787722 TI - How to use teams effectively--and realistically. PMID- 10787723 TI - The economic model for health care delivery. A business plan to create the virtual clinic. AB - This paper reviews briefly the history of managed care, and then analyzes the economic rationale underlying the present dynamics of the marketplace. With this background, the paper postulates what the ultimate delivery system should be: the virtual clinic. It includes a proposed micro-model that could evolve into a macro institution driven by the two essential elements of the market: equity and efficiency. PMID- 10787724 TI - Efficiency and effectiveness: beyond internal practice operations. AB - Physician group practice management teams (physician-administrator teams) have had to become creative in developing efficient and effective practice operations as a result of managed care. For physicians that must use entities outside of the group to deliver care, these physicians must demand that these facilities provide a program that adheres to the principles of "operationally effective and efficient design (OEED)." Programs that comply with the six principles of OEED offer physicians the opportunity to maximize their monetary value of time. Programs that do not follow the OEED principles negatively impact the physicians' monetary value of time. As a result, physician-administrator teams must demand operations improvement from such entities, otherwise alternatives will be sought. Therefore, managed care may not be bad news for physicians. PMID- 10787725 TI - Here to stay: health care compliance. An overview. AB - The strict enforcement of laws governing Medicare fraud and abuse has led to the need for every hospital and other health care facility to develop a compliance program and to update that program periodically. Understanding the history of such enforcement, the regulations involved and the developments on the horizon are vital to avoid huge fines and other severe penalties. Compliance with federal and state regulations will not only avoid such fines and penalties but also will result in a more efficient health care facility. PMID- 10787727 TI - The annual meeting: regulatory obligation or lifeline to the future? AB - Usually dreaded, annual meetings can provide a crucial, powerful, but often neglected context for reviewing the "vital signs" of a group practice: mission, membership, marketplace strategy and infrastructure. We propose that regular and thoughtful attention to essential issues of cohesion and effectiveness significantly increases the likelihood of group vitality and group member satisfaction. We suggest a methodology for embedding these essential conversations into the ritual of the annual meeting. PMID- 10787726 TI - Ensuring patient satisfaction in medical groups. AB - Delivering satisfaction to patients has become increasingly important among professionals in the medical community. However, administrators in medical group practices charged with the task of nurturing customer satisfaction are often required to allocate their limited funds across an array of initiatives intended to ensure the delivery of the right amount and types of services to improve satisfaction among their customers. This requires the ability to locate areas that yield the greatest response per unit of investment. This paper shows that the impact of attribute performance on satisfaction is asymmetric. Positive attribute performance is shown to have a smaller impact on satisfaction than negative attribute performance. The paper also discusses how an understanding of this asymmetry will enable administrators to allocate their resources more wisely as they decide whether to maintain or increase attribute-level performance. PMID- 10787728 TI - Finding meaning in reporting requirements. Provider sampling and survey length. AB - Meeting compliance requirements to report information such as provider satisfaction typically inspires groans, followed by hasty actions by those who must collect and supply the information. Reports often reflect this lack of interest as findings are rarely defended as representative. Our medical group employed statistical methodology to make this process meaningful while also keeping it fairly simple and without incurring added costs. By establishing our provider satisfaction survey as a scientifically based endeavor, we improved the likelihood the provider would actively and honestly participate, and report findings with confidence. PMID- 10787729 TI - Managing the complexities of group practice. An interview with ACMPE president Robert Nelson. AB - Robert A. Nelson, FACMPE, the new president of the American College of Medical Practice Executives (ACMPE) and a principal with the Canon Group, Santa Barbara, Calif., took time to share his vision of the ACMPE and the state of group practice with the MGM Journal. A past president of MGMA. Nelson was honored in 1999 with the ACMPE's Harry J. Harwick Award--the College's highest recognition for health care executives. PMID- 10787730 TI - Goal to reduce errors by 50% in the next 5 years. PMID- 10787731 TI - Century's new challenge: invest in human capital. PMID- 10787732 TI - Acrylic nails harbor pathogens. PMID- 10787733 TI - NIOSH urges use of safer devices. PMID- 10787734 TI - A view of the nursing shortage. PMID- 10787735 TI - A periop course from grassroots. PMID- 10787737 TI - Multi-state license for nursing? PMID- 10787736 TI - Pay for skills aids recruitment. PMID- 10787738 TI - Profile: PAs in surgery. PMID- 10787739 TI - Why try to reduce turnover time? PMID- 10787740 TI - Common ground on peds policies. PMID- 10787741 TI - Benchmarks. Lap chole supply costs show wide variation. PMID- 10787742 TI - Sharps: what will OSHA look for? PMID- 10787743 TI - OSHA proposes ergonomics rule. PMID- 10787744 TI - 'Ticket to work' bill includes fraud unit expansion. PMID- 10787745 TI - IOM report spurs patient safety activity on Capitol Hill. PMID- 10787746 TI - New Jersey strives to give seniors more housing options. PMID- 10787747 TI - Fiscal intermediary settles Medicare misconduct suit. PMID- 10787748 TI - Law calls for study of assisted living programs for veterans. PMID- 10787749 TI - New York facility owner accused of Medicaid fraud. PMID- 10787750 TI - Surviving PPS. PMID- 10787751 TI - Building mission statements that matter. PMID- 10787752 TI - Gimme shelter: OIG's new safe harbors. PMID- 10787753 TI - Covering the costs of a graying America. PMID- 10787754 TI - Achieving 100 percent immunization. PMID- 10787755 TI - 'Poor performing chain' concept dropped. PMID- 10787756 TI - External resources can promote cost management and financial benchmarks for radiology. PMID- 10787757 TI - Securing healthcare applications: digital certificates and PKI. PMID- 10787758 TI - Three-Card Monte and pigs wearing earrings. PMID- 10787759 TI - Project planning, training, measurement and sustainment: the successful implementation of voice recognition. AB - Computerized voice recognition systems (VR) can reduce costs and enhance service. The capital outlay required for conversion to a VR system is significant; therefore, it is incumbent on radiology departments to provide cost and service justifications to administrators. Massachusetts General Hospital (MGH) in Boston implemented VR over a two-year period and achieved annual savings of $530,000 and a 50% decrease in report throughput. Those accomplishments required solid planning and implementation strategies, training and sustainment programs. This article walks through the process, step by step, in the hope of providing a tool set for future implementations. Because VR has dramatic implications for workflow, a solid operational plan is needed when assessing vendors and planning for implementation. The goals for implementation should be to minimize operational disruptions and capitalize on efficiencies of the technology. Senior leadership--the department chair or vice-chair--must select the goals to be accomplished and oversee, manage and direct the VR initiative. The importance of this point cannot be overstated, since implementation will require behavior changes from radiologists and others who may not perceive any personal benefits. Training is the pivotal factor affecting the success of voice recognition, and practice is the only way for radiologists to enhance their skills. Through practice, radiologists will discover shortcuts, and their speed and comfort will improve. Measurement and data analysis are critical to changing and improving the voice recognition application and are vital to decision-making. Some of the issues about which valuable date can be collected are technical and educational problems, VR penetration, report turnaround time and annual cost savings. Sustained effort is indispensable to the maintenance of voice recognition. Finally, all efforts made and gains achieved may prove to be futile without ongoing sustainment of the system through retraining, education and technical support. PMID- 10787760 TI - The current status of positron emission tomography. AB - Positron emission tomography (PET), invented over 25 years ago, is the only imaging technique that provides images of the biological basis of disease. Since disease is a biological process, PET routinely detects disease when other imaging studies, such as CT and MRI, are normal. In addition to its clinical effectiveness, PET has been shown to reduce costs, primarily due to the elimination of other less accurate diagnostic tests and ineffective surgeries. PET has been determined to be applicable to a number of specific applications in the areas of: imaging cancer patients, characterizing myocardial blood flow and viability, and brain imaging in various physiological and pathologic conditions. Tremendous progress has been made in resolving the regulatory and reimbursement issues facing the field of PET. Working with HCFA, representatives of the Institute for Clinical PET and the Society of Nuclear Medicine have brought about expanded HCFA coverage for PET. When HCFA first authorized payment for PET, all coverage decisions were restricted to HCFA and an expanded national coverage policy. HCFA revised its national coverage policy in 1997; this was the first of several steps taken by HCFA towards careful expansion of PET reimbursement. In March 1999, three new indications for whole-body PET scans were added to Medicare's coverage policy. The Institute for Clinical PET is continuing to work with HCFA on continued, appropriate expansion of the coverage policy. This article is partially excerpted from a written statement made by Terry Douglass, Ph.D., president of CTI, Inc., on May 12, 1999, before the Senate Committee on Commerce, Science and Transportation and its Subcommittee on Science, Technology and Space. This was part of the committee's study of "Emerging Technologies in the New Millennium." PMID- 10787761 TI - Positron emission tomography: a first-hand experience. AB - In July 1999, the University of Kansas Hospital installed a positron emission tomography (PET) scanner and added PET to the imaging technologies it offers patients and physicians. The new service is managed by the nuclear medicine section in the department of radiology. Plans are being implemented now to install a cyclotron in March 2000. Prior to installation of the scanner, a radiation area survey was performed in the space being considered for the PET unit. We also needed to address other critical considerations, including the manufacturer's requirements for construction of the scanner room, special electrical needs, and how the system would connect to our existing information network. It is important to work closely with your chief financial officer and chief operations officer from the beginning of the purchasing process so that these administrators have up-to-date, supportive information about PET and the progress of the installation. We made use of a variety of promotional techniques to market the new service, including broadcast e-mail, an open house for potential referring physicians, postings on the nuclear medicine Web site and communication through the local media. We also worked with the major insurance providers that utilize our hospital to educate them about PET and its benefits. In addition, we trained our own billing staff about procedures that optimize reimbursement for PET. In March 2000, University of Kansas Hospital will install the first cyclotron in the state, enabling us to generate the drugs used for PET scanning and potentially to add targets for research PET radiopharmaceuticals. PMID- 10787762 TI - A new approach to the film library: time-unit filing. AB - The installation of a new radiology information system (RIS) at Children's Hospital Medical Center of Akron in Akron, Ohio, took the radiology department into a new world of technology, but raised issues we never anticipated. The major problem the new RIS forced the department to overcome was how to eliminate the film file's reliance on a proprietary radiology numbering system. Previously, the department had used its own numbering system--a proprietary x-ray number--to file film jackets and had used the hospital-issued medical record number to access patient and payer information from the hospital information system. It became clear that we should use a single number--the medical record number--to access all data, but we wondered how that would affect our film file room. An RIS consultant suggested that we consider filing films by last date of service, a system called "time-unit filing." Time-unit filing means keeping the most recent two-weeks worth of films in the main file room. They are organized by gender in blue or pink jackets and marked alphabetically by the patient's last name in a way that makes mis-files easy to see. If a patient's film jacket is activated again, it is refiled in the current two-week time unit. Inactive jackets remain in their two-week time unit indefinitely. Time-unit filing has had many benefits for the radiology department at Children's Hospital Medical Center of Akron: fewer mis-files, less time needed for filing and searching, and successful implementation of the new RIS. PMID- 10787764 TI - I want it, and I want it now! PMID- 10787763 TI - The use of alternative and complementary medicine in radiology today. AB - There is a quiet revolution challenging our Western philosophy of reactive, high tech allopathic medicine, which cures disease by treatments. This trend to a proactive, preventive, Eastern style of medicine is referred to as holistic or alternative and complementary medicine (ACM). ACM is based on the belief that human beings are living energy systems rather than an arrangement of parts. Any disturbance in the body, mind or spirit will result in a disturbance in the whole system. In January 1993, the New England Journal of Medicine published a study that documented the rise of ACM in the United States. Expenditures associated with the use of unconventional therapies in 1990 amounted to approximately $13.7 billion, three quarters of which ($10.3 billion) was paid for out of pocket. ACM techniques that are currently used in diagnostic imaging departments include guided imagery, anodyne imagery, aromatherapy and hydrotherapy. Radiology can play a crucial role in moving beyond allopathic and complementary medicine to a combination approach. PMID- 10787765 TI - PETs scanned here. PMID- 10787766 TI - The effects of an educational intervention on calcium intake and bone mineral content in young women with low calcium intake. AB - PURPOSE: This study evaluated whether a combined behavioral and dietary intervention would affect young women's calcium intake and bone mineral content (BMC). DESIGN: The design was a two by three mixed design with one between subjects factor (treatment vs. control) and one within-subjects factor (time- baseline, 3-month, and 6-month). SETTING: The study was conducted in a university setting in Memphis, Tennessee. PARTICIPANTS: A total of 80 premenopausal women (ages 18 to 30) with low baseline calcium intake (< 700 mg/d) were included in the analyses. There were 40 women in the treatment group and 40 women in the control group. MEASURES: Hertzler and Frary's rapid assessment questionnaire was employed to evaluate calcium intake, and dual-energy x-ray absorptiometry (DEXA) was employed to assess BMC. RESULTS: Repeated measures analysis of variance (RM ANOVA) was employed to analyze results. Results indicated that women in the treatment group made greater increases in total calcium intake and supplemental calcium than women in the control group and that all women made significant increases in dietary calcium intake. Additionally, analyses of BMC revealed that women in the treatment group did not experience significant changes in total BMC, and women in the control group experienced significant losses in total BMC. CONCLUSIONS: In the current study, women were losing BMC, and the women who made the largest increases in calcium intake were able to retard this bone loss. There is increasing evidence that dietary calcium intake in young people is extremely low, and the results of the current study highlight the need for much more intensive evaluations investigating the factors that are positively associated with premenopausal bone mineral change. PMID- 10787767 TI - How well do consumers understand percentage daily value on food labels? AB - One hundred four adults completed a multiple choice food label comprehension questionnaire and a food frequency questionnaire. Most respondents were able to accurately rate the fat content of five different foods using only the food label information. However, some underestimated food content for some foods. Also, only 29% correctly selected the definition of % daily value for fat (%DV), as "percent of the maximum daily recommended amount of fat." PMID- 10787768 TI - Effects of educational materials concerning osteoporosis on women's knowledge, beliefs, and behavior. AB - PURPOSE: This study examined the effects of brief written educational materials on osteoporosis-related knowledge, beliefs, and behaviors. The study also examined whether observed effects varied as a function of one's stage in the precaution adoption process. DESIGN: The study used an experimental research design. SETTING: Participants were identified from North Carolina driver's license records. SUBJECTS: Of the 1476 women in the initial sample, 536 (36.3%) enrolled in the study and 307 completed all follow-up assessments. INTERVENTION: Participants were randomly assigned to one of four groups. One group received an information packet containing general information about osteoporosis. One group received an action plan packet containing instructions on how to increase one's level of exercise and calcium intake. One group received both packets. The final group received neither packet. MEASURES: Primary study variables were beliefs related to osteoporosis, calcium, and exercise; osteoporosis knowledge; calcium and exercise stage; calcium intake; and exercise level. RESULTS: Overall, receipt of the information packet was associated with changes in knowledge and beliefs (F[18,283] = 2.11, p < .01) irrespective of participants' stage of change. No effects on behavior were observed. CONCLUSIONS: These findings suggest that brief written educational materials can facilitate knowledge and belief change but that they do not promote behavior change. The generalizability of these findings is limited by the low study response rate. PMID- 10787769 TI - Refining a tool to measure cues to action in encouraging health-promoting behavior--the CHAQ. AB - A convenience sample of 99 questionnaire respondents was used to develop an instrument to measure "cues to action" for health-promoting behaviors. The questionnaire contained 32 items producing an alpha coefficient of 0.88. Because of the small sample size and limitations of the convenience sample, the authors recommend that their findings be considered preliminary. PMID- 10787770 TI - Private sector worksite health promotion programs in Missouri: a comparison with national survey findings and Healthy People recommendations. AB - Questionnaries with 22 multiple-choice questions were mailed to a random sample of 450 Missouri employers with 250 or more employees to determine the prevalence and types of workplace health promotion programs; 262 (58%) responded. Only 34% of the responding companies offered any kind of program and 65% of these offered only awareness programs, but most offered programs on all 18 topics listed in the questionnaire; 48% had a full-time program manager, 79% had some form of evaluation, and 36% had some form of financial incentives. PMID- 10787772 TI - Going beyond the usual suspects. PMID- 10787771 TI - Process tracking results from the Treatwell 5-a-Day Worksite Study. AB - PURPOSE: To report findings from Treatwell 5-a-Day process tracking. DESIGN: Worksites were randomly assigned to a minimal intervention control, worksite-only condition, or worksite-plus-family condition. SETTING: Twenty-two small community health centers in Massachusetts. SUBJECTS: Employees of the community health centers. INTERVENTION: Both intervention conditions included the formation of employee advisory boards; activities such as nutrition discussions and taste tests targeting individual behavior change; and point-of-purchase labeling as an environmental strategy. Worksite-plus-family sites incorporated activities such as family contests, campaigns, and picnics. MEASURES: Documentation of the number and type of activities for extent of implementation; number of participants in activities for reach; program awareness and participation from the follow-up employee survey (n = 1306, representing 76% [range, 56%-100%] of the sample); change in fruit and vegetable consumption from a comparison between the follow-up and baseline surveys (n = 1359, representing 87% [range, 75%-100%] of the sample). RESULTS: A higher number of activities per employee was significantly correlated with greater program awareness (.68; p = .006) and greater change in fruit and vegetable consumption (.55; p = .04). Greater participation in activities was significantly correlated with greater awareness (.67; p = .007), higher participation (.61; p = .02), and increase in fruit and vegetable consumption. (.55; p = .04). CONCLUSIONS: These results provide quantitative indicators of a dose-response relationship between the number of intervention activities per employee and higher percentage of employee participation and observed increases in fruit and vegetable consumption. PMID- 10787773 TI - A framework for building effective public health constituencies. AB - Population-based health improvements that require behavioral and social change at the community level are dependent on effective constituency participation. To achieve needed constituency involvement, a public health leader must understand what motivates and moves constituents to action on public health issues. This article provides a framework and guidance on building effective constituent involvement to achieve community health improvement. Within this framework, aspects of managing the organizational practice of constituency building and community engagement are discussed and linked with current public health planning and mobilization models that support community-based health interventions. PMID- 10787774 TI - The implementation and evaluation of a healthy communities process in central Alberta: some implications for public health practice. AB - This article describes the implementation and evaluation of a Healthy Communities Initiative (HCI) by the David Thompson Health Region in central Alberta, Canada. The HCI model provided for a facilitated, community-based, strategic planning process. Its key steps include development of a shared vision of health, assessment of needs and strengths, selection of key priority areas for action, and implementation of strategies to achieve change. A three-level evaluation model was developed, which incorporates project-level evaluation, cluster-level evaluation, and critical reflection on the David Thompson Health Region's own capacity to engage in community development work. PMID- 10787775 TI - Community network linkages during a health controversy. AB - The communication of complex scientific and technical health risk information in a community context is little understood and seldom studied. The study reported here examines how information about an environmental health controversy is obtained, exchanged, and used within the community as decision makers and public health officials struggle to protect lake-source drinking water for a community of 55,000. Findings suggest that public health officials may need to broaden their communication strategies to reach less integrated groups with quality scientific and technical health risk information. PMID- 10787776 TI - County commissioners as a key constituency for public health. AB - Local government elected officials are an important constituency for public health as they represent all citizens in their cities and counties, make policy decisions that affect the public's health, and allocate funding to support public health activities. This article describes efforts to determine the current knowledge and perspective of county commissioners regarding factors that affect local governmental decision making, particularly involving public health. Interview findings indicate that county commissioners are influenced by what their constituents consider important. The article suggests actions that public health managers can take to increase local elected officials' understanding of public health. PMID- 10787777 TI - The changing face of local public health: a case example. AB - Local public health agencies are looking for new ways to position themselves within the rapidly changing health care environment. This article provides background on one community's process for developing an innovative, collaborative organizational model for delivering public health and primary care services. Challenges and opportunities, key stakeholders, and transition issues are highlighted. PMID- 10787778 TI - State public health laws: an expression of constituency expectations. AB - One expression by the body politic about public health in any jurisdiction is the law under which the public health agency practices. The degree of congruence between a widely used model of public health and current statutes can be a measure of that opinion. This study identified the extent to which the statements of purpose expressed in state laws are consistent with Public Health in America (PHA); the published mission statements of state health agencies are consistent with either statute or PHA; and which concepts from PHA are most underrepresented and what additional concepts are included. PMID- 10787779 TI - Coalition, partnership, and constituency building by a state public health agency: a retrospective. AB - This article is a retrospective that traces the development of an evolutionary process for a state health agency in addressing the challenge of implementing core public health functions and the provision of essential services. Coalition, partnership, and constituency building were critical elements in the process. Various initiatives are described and their importance as building blocks to a larger process of organizational change is explained. Key lessons from the process are outlined. PMID- 10787780 TI - Building constituencies to promote health: a case study. AB - In the practice of public health today, a greater emphasis is being placed upon working collaboratively with many community partners. This creates more opportunities to measurably improve community health. The benefits and challenges of establishing constituencies to promote health in the capital region of New York State are presented in this article. PMID- 10787781 TI - Developing a framework for comprehensive cancer prevention and control in the United States: an initiative of the Centers for Disease Control and Prevention. AB - The Division of Cancer Prevention and Control, Centers for Disease Control and Prevention, is working with state health agency staff and other stakeholders to develop a comprehensive and integrated approach to cancer control. To help stakeholders visualize the approach, a graphic model was developed based on stakeholder input and a literature review of existing models. Phases of the model include setting optimal objectives (data driven), determining optimal strategies (science driven), establishing feasible priorities (capacity driven), and implementing effective strategies (outcome driven). The model currently is being validated through case studies of state-level cancer planning in six states. PMID- 10787782 TI - Comprehensive cancer control initiative of the Centers for Disease Control and Prevention: an example of participatory innovation diffusion. AB - Site-specific and risk factor-specific cancer programs can point to impressive accomplishments, but coordination among them often is lacking. The Division of Cancer Prevention and Control, National Center for Chronic Disease Prevention and Health Promotion, Centers for Disease Control and Prevention, is working with state health agency staff and other stakeholders to develop a comprehensive, integrated, nationwide approach to cancer control. The participatory innovation diffusion model may help this complex public health innovation be adopted. The participants in the process identified problematic aspects of the innovation and steps that the division can take to ameliorate these problems before the innovation is implemented. PMID- 10787783 TI - School-based asthma management policies and practices: a regional survey. AB - Children with asthma may spend as much as one-third of their weekdays in the school setting. As one effort to evaluate and improve asthma care in the Dallas/Fort Worth area, the North Central Texas HEDIS Coalition surveyed 36 independent school districts (ISDs). Twenty-nine ISDs, representing 679,631 children, responded to questions on school health personnel, identification and monitoring of students with asthma, staff training in asthma care, policies for students carrying asthma medications, and campus smoking policies. Key findings were more positive than expected and included the following: (a) training in asthma management is fairly extensive, (b) students in 26 ISDs are allowed, upon physician request, to carry medihalers, and (c) smoking on school property is illegal in Texas. However, the absence of a fully developed strategy for implementing the survey findings may have hindered the continuous quality improvement efforts among schools, health plans, providers, and the community that could have evolved from this project. PMID- 10787784 TI - Hitting the mark in community health outreach: what counts? AB - This article describes a nurse-managed community health outreach project, at the Health Promotion Center (HPC) of Fairfield University School of Nursing, to prevent cardiovascular disease in an inner-city community. Flexibility in the project's grant funding has enabled the HPC to use a customer-oriented approach to develop programs. The article details the application of community health nursing and quality management practices to the development of HPC's cardiovascular-disease-screening program. PMID- 10787785 TI - The performance measurement coordinating council: a town meeting. AB - A Town Meeting about the Performance Measurement Coordinating Council (PMCC) was held at the NAHQ Annual Educational Conference in September in Atlanta. Established in 1998, the PMCC is a collaboration of three national accrediting bodies--the American Medical Accreditation Program (AMAP), the Joint Commission on Accreditation of Healthcare Organizations (JCAHO), and the National Committee for Quality Assurance (NCQA)--to coordinate performance measurement activities among the three organizations and across the healthcare system. Representing the Joint Commission was Dennis O'Leary, MD; representing AMAP was Yank Coble, MD; and representing NCQA was Joshua Seidman, MS BS. This article is a synopsis of that meeting, prepared from the actual transcript. The meeting began with each speaker's opening remarks, followed by questions from the audience. PMID- 10787786 TI - Clinical practice guideline for managing acute urinary tract infections in adult females in Saudi Arabia. AB - The management of urinary tract infections at primary health clinics in Saudi Arabia varied widely and did not generally conform to modern recommendations. A clinical practice guideline was developed to standardize and modernize the approach in order to improve the quality of care and reduce costs. An algorithm was developed for the management of acute urinary tract infections in non pregnant adult females. Guideline recommendations were generally well received, and acceptance was greater where users were involved in the initial stages of guideline development. The cost of managing urinary tract infections was reduced providing that reliance on dipstick analysis did not increase costs through increased misdiagnosis. PMID- 10787787 TI - A users group approach to quality improvement across an integrated healthcare delivery system. AB - Aurora Healthcare, an integrated healthcare delivery system, wanted to use interhospital collaboration to improve quality. An opportunity for improvement at five Aurora hospitals was identified: Only 71% of patients received timely preoperative antibiotic prophylaxis. A Users Group approach was proposed to foster collaboration among the hospitals in a quality improvement (QI) effort. Although these hospitals differed in size, volume of cases, medical staffs, and antibiotic-delivery processes, they had common objectives and, many times, common barriers to the timely delivery of antibiotics. A Users Group approach to this QI initiative resulted in an 18% improvement in the number of patients who received timely administration of preoperative antibiotic prophylaxis (p < 0.001). This article describes the Users Group approach to QI at Aurora Healthcare in southeastern Wisconsin. PMID- 10787788 TI - On the future of quality in healthcare. PMID- 10787790 TI - Turning data into consumable information. PMID- 10787789 TI - Dear healthcare quality professionals:. PMID- 10787791 TI - Economic links between cardiovascular groups and hospitals: a new approach. AB - Recent regulatory developments complicate the process of aligning the interests of cardiovascular groups with their hospital partners. However, the need for alignment has never been greater. Declining revenues make it essential that physicians and hospitals cooperate in every way to reduce cost while at the same time preserving value to patients. The CIB Joint Venture concept described in this article provides a low-risk option that is worth consideration. PMID- 10787792 TI - Turning negative feedback from business offices into positive results. PMID- 10787793 TI - Disease management: making the build buy decision. PMID- 10787794 TI - Health-care systems and ethics: what can we learn? AB - Health care systems in different countries and cultures differ and tend to reflect the particular values and, therefore, the particular social structure of a given society. Each of these has ethical problems unique to itself. Some of these problems are briefly discussed. So as to have an individual ethical problem in the context of medical care, access to medical care needs to be assured. It is argued that individual problems are the primary issue in societies in which there is fair access whereas they are of lesser importance in societies which have thus far failed to provide fair access. PMID- 10787796 TI - Some aspects of the reform of the health care systems in Austria, Germany and Switzerland. AB - The health care systems in Austria, Germany and Switzerland owe their institutional structure to different historical developments. While Austria and Germany voted for the Bismarck-Model of social health insurance, Switzerland adopted a voluntary system of health insurance. In all three countries, until very recently, the different challenges which the health care sector faced were met by piecemeal approaches and by stop and go policies, which, in the long run were not very successful either in containing costs or in improving efficacy and efficiency. During the 1990 more fundamental reforms in the health care systems of all three countries took place. Germany and Switzerland chose the path of deregulation of the health insurance system, which consequently strengthened the competition between the insurance companies, and, to some extent between the suppliers of medical services. While this can be seen as an essential part of the reform process for these two countries. Austria favors a state-oriented and interventionist approach in order to meet the challenges. PMID- 10787795 TI - Access to health care in the Scandinavian countries: ethical aspects. AB - The health care systems are fairly similar in the Scandinavian countries. The exact details vary, but in all three countries the system is almost exclusively publicly funded through taxation, and most (or all) hospitals are also publicly owned and managed. The countries also have a fairly strong primary care sector (even though it varies between the countries), with family physicians to various degrees acting as gatekeepers to specialist services. In Denmark most of the GP services are free. For the patient in Norway and Sweden there are out-of-pocket co-payments for GP consultations, with upper limits, but consultations for children are free. Hospital treatment is free in Denmark while the other countries use a system with out-of-pocket co-payment. There is a very strong public commitment to access to high quality health care for all. Solidarity and equality form the ideological basis for the Scandinavian welfare state. Means testing, for instance, has been widely rejected in the Scandinavian countries on the grounds that public services should not stigmatise any particular group. Solidarity also means devoting special consideration to the needs of those who have less chance than others of making their voices heard or exercising their rights. Issues of limited access are now, however, challenging the thinking about a health care system based on solidarity. PMID- 10787797 TI - Health care in France: recent developments. AB - Health care in France falls almost exclusively under the responsibility of the Social Security department, which covers almost all the expenditures related to health care, whether hospitalization or medication is concerned. For severe diseases or surgery the coverage is likely to reach as much as 100%. The medical expenditures for several severe diseases, such as cancer, myocardial infarction, or neurodegenerative diseases are 100% covered for a period of time as long as three months. For some procedures, full coverage may be achieved by using a subscription to private health care insurance. Access to cover by the state has recently been opened to anyone living in France, after passage of a special law. There is still a lack of intensive care and hospice beds, given the rapidly increasing number of elderly who cannot be maintained at home. There is a tendency to reduce the number of beds in private and public hospitals due to the great number of such institutions and the general concern that a low volume of procedures, associated with inexperienced health care professionals, is likely to increase morbidity and mortality as well as public health care expenditure. Patients are still free to choose doctors and hospitals (whether private or public), provided that beds are available and that specific procedures can be carried out in the vicinity. So far no waiting list is needed, except for specific procedures performed by a few specialists of high repute. Health care expenses are increasing continuously, which results in a very expensive system in France. PMID- 10787798 TI - The British National Health Service: a tarnished moral vision? AB - Last year (1998) saw the celebration of the 50th Anniversary of the British National Health Service (NHS). One of the few completely nationalized systems of health care in the world, the NHS is seen by many as a moral beacon of what it means to provide equitable medical treatment to all citizens on the basis of need and need alone. However, others argue that it has failed to achieve the overall goals for which it was created. Because of scarce resources, some urgently needed care is not available at all, while that which is received is sometimes second class. For these reasons, it is claimed that the NHS should be scrapped and replaced by other systems of health care delivery. This paper outlines the history of the NHS, indicating some of the problems and innovations which have led to its current organization and structure. The philosophical foundations of the NHS are then articulated and defended on the grounds that it still represents a morally coherent and economically efficient approach to the delivery of health care. Scarce resources are the key problem facing the NHS, making rationing inevitable and it is shown that this is not incompatible with the moral foundations of the service. However, there can be little doubt that the NHS is now becoming dangerously under-funded. The paper concludes with arguments about why this is so and what might be done about it. PMID- 10787799 TI - The Canadian health care system. An analytical perspective. AB - The Canadian health care system is a publicly funded system based on the philosophy that health is a right, not a commodity. The implementation of this perspective is hampered by the fact that the Canadian Constitution makes health care a matter of provincial jurisdiction, while most taxing powers lie in the hands of the federal government. Further problems arise because of Canada's geographic nature and a move to regionalization of provincial health care administration. The issue is compounded by recent developments in reproductive technologies, aboriginal health, changes in consent law, etc. PMID- 10787800 TI - The United States health care system under managed care. How the commodification of health care distorts ethics and threatens equity. AB - Describing the U.S. health care system means describing managed care under commercial forces. Managed care creates new moral tension for practitioners, but more importantly, in its current form it intensifies the commercialization of health expectations and interactions. The largely unregulated marketing of health services under managed care has been a major factor in the increasing number of uninsured citizens, while claims for cost reduction through managed care are equivocal. Risk-rating practices integral to the current medical marketplace thwart concerns for justice in allocation and create vulnerabilities for almost everyone. The political-moral concern of the early 1990s for a right to health care is nowhere in sight. PMID- 10787801 TI - HCFA issues more instructions on Balanced Budget Refinement Act. PMID- 10787802 TI - HCFA modifies requirements on physician certification for nonemergency ambulance services. PMID- 10787805 TI - SNFs can manage risk in today's perilous environment by focusing on quality care. PMID- 10787804 TI - AMDA white paper identifies ways to improve pharmaceutical care in SNFs. PMID- 10787803 TI - New Medicare Part B therapy fee schedules in effect. PMID- 10787806 TI - Jefferson Health System on path to improved clinical performance through report cards. PMID- 10787808 TI - Community hospitals rush to specialize in tight market. PMID- 10787807 TI - Sentara Medical Group goes back to the future. PMID- 10787809 TI - Managed care tactics can "work" in workers' compensation. PMID- 10787810 TI - Hospitals discover cost efficiency of private rooms. PMID- 10787811 TI - Turning medical error tragedy into opportunity. PMID- 10787812 TI - Into the mix results from PPS per-episode demonstration & case-mix study. AB - The Health Care Financing Administration (HCFA) released two reports in October 1999 that provide insight into the development of a per-episode prospective payment system (PPS) for Medicare home health. These reports provide important clues about two critical issues: the impact of per-episode payment on the cost of providing home health care, and how to predict the resources used by home health patients. PMID- 10787813 TI - Developing a case-mix model for PPS. AB - Agencies are pinning hopes for success under PPS on an accurate case-mix adjustor. The Health Care Financing Administration (HCFA) tasked Abt Associates Inc. to develop a system to accurately predict the volume and type of home health services each patient requires, based on his or her characteristics (not the service actually received). HCFA wanted this system to be feasible, clinically logical, and valid and accurate. Authors Goldberg and Delargy explain how Abt approached this daunting task. PMID- 10787814 TI - Ask the experts ... the upcoming shift to a prospective payment system (PPS). PMID- 10787815 TI - Agencies at risk: increased liability under PPS. AB - With the long anticipated shift from cost reimbursement to a prospective payment system (PPS) for Medicare home health services, the home care industry can look forward to improved opportunities for service flexibility, the end to retroactive cost disallowances, and opportunities for financial rewards through efficiencies in operation. However, bringing home care into a more modern world of reimbursement also means that new forms of risk emerge, creating the potential for a liability that must be recognized and properly managed. PMID- 10787816 TI - Leveraging health care processes using technology applications. AB - Providers are being encouraged to deliver health care services differently, and use resources more efficiently while maintaining or improving patient outcomes and satisfaction. Agencies should understand how to use information technology in leveraging the health care industry's ability to improve access to care, facilitate collaboration in delivering care and health information, and decrease costs related to the rendering of care. PMID- 10787817 TI - OIG's compliance guidance for hospices. PMID- 10787818 TI - New Home Care Aide Code of Ethics focuses on client-centered quality care. PMID- 10787819 TI - Becoming a virtual caregiver. AB - The home care community is becoming increasingly familiar with the concept of telehealth services for the home, and perhaps more receptive to using such services. Telehealth services might fit well within the health care continuum as an "adjunct" to conventional services. PMID- 10787820 TI - Reaching out ... through communication. PMID- 10787821 TI - PPS policy decisions have direct impact on providers and patients. PMID- 10787822 TI - Welcoming the century of caring. PMID- 10787823 TI - Home health PPS: new payment system, new hope. AB - In light of the financial turmoil most home health providers experienced over the past two years, a new payment system brings a sense of hope. The prospective payment methodology will provide different incentives than a cost-based reimbursement system. These new incentives will open the way for new approaches to deliver home health care. However, this new hope is accompanied by a sense of anxiety because the system is new and untested. PMID- 10787824 TI - Examining and embracing the value of coalition participation. PMID- 10787825 TI - Health care cost trends ... and implications for employer benefit strategies. PMID- 10787826 TI - In defense of Social Security. PMID- 10787827 TI - The searchers: how consumers can find cost-effective, quality health care. PMID- 10787828 TI - COBRA: managing the new problems. PMID- 10787829 TI - Workplace strategies for removing obstacles to employee health. PMID- 10787830 TI - Balancing the books. A guide to the budget process for nonfinancial managers. PMID- 10787831 TI - Take note! Thorough documentation of patient care may prevent legal wranglings. PMID- 10787832 TI - Dial "E" for Everest. World's highest mountain serves as base for high-tech medical research. PMID- 10787834 TI - Paramedics in the ED. PMID- 10787833 TI - Buckle up! February marks Child Passenger Safety Awareness Week. PMID- 10787835 TI - Paramedics in the ED. PMID- 10787836 TI - Paramedics in the ED. PMID- 10787837 TI - Paramedics in the ED. PMID- 10787838 TI - Paramedics in the ED. PMID- 10787839 TI - Paramedics in the ED. PMID- 10787840 TI - Paramedics in the ED. PMID- 10787841 TI - Paramedics in the ED. PMID- 10787842 TI - Update to HIV & AIDS article. PMID- 10787843 TI - Medicare and Medicaid fraud. PMID- 10787845 TI - Compliance in the organizational ethics context. PMID- 10787844 TI - Above reproach: developing a comprehensive ethics and compliance program. AB - How can a healthcare organization improve the public's confidence in the conduct of its business operations? What can it do to ensure that it can thrive despite being the subject of public and governmental scrutiny and doubt? Healthcare providers must establish standards of conduct that are above reproach and ensure that those standards are clearly articulated and strictly adhered to. This article describes the merits of a comprehensive ethics and compliance program, suggests five basic elements of such a program--organizational support/structure, setting standards, creating awareness, establishing a mechanism for reporting exceptions, and monitoring and auditing--and then demonstrates how those elements should be applied in several high-risk areas. Fundamentally, an ethics and compliance program has two purposes: to ensure that all individuals in an organization observe pertinent laws and regulations in their work; and to articulate a broader set of aspirational ethical standards that are well understood within the organization and become a practical guideline for organization members making decisions that raise ethical concerns. Every ethics and compliance program should contain certain fundamental aspects. First, the effort must have the active support of the most senior management in the organization. To instill a commitment to ethics and compliance absent a clear and outspoken commitment to such purposes by organization leaders is simply impossible. Second, an ethics and compliance program is fundamentally about organizational culture--about instilling a commitment to observe the law and, more generally, to do the right thing. Third, ethics and compliance are responsibilities of operating management (sometimes called line management). Although staff such as compliance officers are obligated to provide the necessary resources for a successful program and to design the program, such staff officers cannot achieve implementation and execution. Only operating managers can do that. Fourth, an ethics and compliance effort should be about the conduct of individuals, not about "checking the boxes" in a model plan or generating attractive written or educational materials. Such an effort is about individuals on a day-to-day basis knowing what is expected of them and doing it and about never compromising integrity, regardless of pressures faced. A great deal of progress has been made in healthcare organizations in the development of increasingly sophisticated ethics and compliance programs. A particularly energetic focus has been placed on these programs since formal government guidance regarding compliance programs was first issued in the laboratory area about two years ago and as more sophisticated automated monitoring tools have been developed. As ethics and compliance programs have become more sophisticated, certain best practices have been established. This discussion will set forth approaches to ethics and compliance in the context of what are believed to be illustrative best practices. Much of what is described here is descriptive of the efforts of Columbia/HCA Healthcare Corporation from October 1997 to the present; however, this article has been presented not as a mere descriptive piece but rather as a set of normative guidelines. We hope that other healthcare providers will find this to be of practical use. Provider settings pose certain unique challenges that are specifically addressed in this discussion; however, many of the issues raised can be adapted to other healthcare organizations. For simplicity's sake, because the authors of this article all work on a daily basis primarily with hospitals, the article is written from a hospital perspective. PMID- 10787846 TI - Doing things right. PMID- 10787847 TI - The compliance program: an application. PMID- 10787848 TI - Media misses the mark. PMID- 10787849 TI - EMS providers nationwide face lawsuits. PMID- 10787850 TI - Avoid preconceived notions. PMID- 10787851 TI - 20 trauma traps to avoid. Methods to expedite the care & transport of the trauma patient. PMID- 10787852 TI - Is MAST in the past? The pros and cons of MAST usage in the field. AB - Initially, MAST was viewed as a panacea for hypovolemic trauma patients. Through decades of study, this has not panned out. However, MAST seems to stabilize and decrease bleeding in pelvic and long-bone fractures of the lower extremities. It also appears useful in anaphylaxis and in non-traumatic intra-abdominal hemorrhage. The suit has proven deleterious to trauma victims with moderate hypotension (systolic BP 50-90 mm Hg) who face only a short ride to a hospital, especially those with thoracic injuries. Its role in patients with severe hypotension or long prehospital transport times remains unclear. In severely hypotensive patients, the improvement in BP and oxygenation to the heart and brain may override any negative effects of continued hemorrhage. We must wait for further studies to resolve these issues. PMID- 10787853 TI - Medically directed use of MAST. PMID- 10787854 TI - Goin' the distance. Endurance sports present an assessment dilemma for EMS personnel. PMID- 10787855 TI - 1999 200-city survey. EMS trends in America's most populous cities. PMID- 10787856 TI - RSV not the common cold. Signs & symptoms of respiratory syncytial virus. PMID- 10787857 TI - Tamponade testimony. PMID- 10787858 TI - Minions of mercy. PMID- 10787859 TI - Medical education has become an assembly line. Interview by Robert Lowes. PMID- 10787860 TI - How much are groups paying their doctors? PMID- 10787861 TI - The man with a 3-inch nail in his head. PMID- 10787862 TI - Should doctors prescribe religion?. Interview by Anita J Slomski. PMID- 10787863 TI - Robert McCool, MD opening doors to the uninsured. PMID- 10787864 TI - If hospital policy jeopardizes a patient. PMID- 10787865 TI - I wish I'd fought my malpractice suit. PMID- 10787866 TI - Now what? 7 tasks for a new beginning. PMID- 10787867 TI - Your own five-year plan: what do you want to accomplish? PMID- 10787868 TI - The challenge for each of us: finding a way to achieve our best. PMID- 10787869 TI - Could a "coach" get you motivated about medicine? PMID- 10787870 TI - Still practicing after all these years. PMID- 10787871 TI - Would you want your kids to follow in your footsteps? PMID- 10787872 TI - Want to uncover a patient's real problem? BATHE him! PMID- 10787873 TI - How to stay sane in the crazy world of medicine. PMID- 10787874 TI - Everything I need to know about medicine I learned as a senior medical student. PMID- 10787875 TI - "Take two worts, and call me in the morning". PMID- 10787876 TI - Re-engineer your practice--starting today. PMID- 10787877 TI - Skeptical reaction to UnitedHealthcare's "we trust doctors" gambit. PMID- 10787878 TI - It's a tougher go, but doctors are still wringing out profits. PMID- 10787879 TI - The biggest problem in American health care. PMID- 10787880 TI - Congress takes some bite out of the 1997 reforms. PMID- 10787881 TI - The California nightmare: is this where managed care is taking us? PMID- 10787882 TI - What's this IPA doing right? PMID- 10787883 TI - Pay debts and save for retirement--on a fellow's stipend? PMID- 10787884 TI - Doctor unions: time to join the parade? PMID- 10787885 TI - Warm-hearted care--or cold-hearted rationing? PMID- 10787886 TI - Medical records privacy gets new push. PMID- 10787887 TI - ADH-associated pathologies. Diabetes insipidus and syndrome of inappropriate ADH. PMID- 10787888 TI - Music medicine. PMID- 10787889 TI - Human resources planning. Building a case for cross-training. PMID- 10787890 TI - Radio frequency technology in the lab. PMID- 10787891 TI - Preventing mishaps team effort, error expert says. PMID- 10787893 TI - Benchmarks. Study indicates fewer mastectomies being performed. PMID- 10787892 TI - How ORs manage on-call varies by local market. PMID- 10787894 TI - RN seeks answers in father's death. PMID- 10787895 TI - Marking site for correct surgery. PMID- 10787896 TI - Top 100 hospitals thrive in lean times. PMID- 10787897 TI - ORs moving to flat-rate charges. PMID- 10787898 TI - Good costing is key to flat charges. PMID- 10787899 TI - Help for missing-in-action devices. PMID- 10787900 TI - FDA moving on reuse regulation. PMID- 10787901 TI - Community newsletters on the Web. AB - An in-depth look at how several hospitals incorporate community newsletters on their Web sites. Featuring St. Luke's Hospital, Sioux City, Iowa; Advocate Health Care, Oak Brook, Ill.; Greenwich Hospital, Greenwich, Conn.; and more. PMID- 10787902 TI - Rancho creates huge turnaround using marketing strategies. The marketing program nabs awards, but the work captures hearts. PMID- 10787903 TI - Research leads to smooth switch for health system's corporate identity. PMID- 10787904 TI - Preparation and improvisation keep North Carolina hospitals afloat after Hurricane Floyd. AB - Hurricane Floyd leaves North Carolina in the wake of a state-wide disaster. Pitt County Memorial Hospital, Greenville, N.C., survives by relying on its tried-and true disaster plan with a little improvisation mixed in. Also, how the North Carolina Hospital Association is pitching in. PMID- 10787905 TI - St. Cloud Hospital works hard to build relationships between its cardiac facility and area physicians. PMID- 10787906 TI - Hospital's physician orientation smoothes service to patients, staff relationships. AB - Fairfield Medical Center, Lancaster, Ohio, put in place a new orientation program for incoming physicians. As a result, many hassles of getting acquainted with a workplace have been smoothed over. PMID- 10787907 TI - Research critical element for successful PR plan. AB - The Phoenix VA Hospital greatly boosts its image in the community, thanks to an aggressive marketing campaign. Staff morale has also gone up due to the success of the program. PMID- 10787908 TI - Women's hospital extends fund-raiser lecture series to be a year-long event. After 10 years, it was time to shake up the program. AB - Magee-Womens Hospital, Pittsburgh, takes an established fundraiser and breathes new life into it. The two-day "Celebration of Women" is now a year-long lecture series, and the results so far are promising. PMID- 10787909 TI - Promotion of osteoporosis awareness drives traffic to hospital library. PMID- 10787911 TI - Republicans unveil BBA refinement bills. PMID- 10787910 TI - University of Iowa provides the definitive institutional Web site. PMID- 10787912 TI - Caregivers march on Washington to support PPS fix. PMID- 10787913 TI - Federal fraud busters eye Medicare contractors. PMID- 10787915 TI - Medicare providers still not ready for Y2K. PMID- 10787914 TI - Nursing facility residents, staff survive Floyd's fury. PMID- 10787916 TI - Vencor files for bankruptcy, employees will be paid. PMID- 10787917 TI - Seniors housing is underappreciated as an investment. PMID- 10787918 TI - How will long-term care remember the Clinton years? PMID- 10787920 TI - Financial services suppliers. PMID- 10787919 TI - AHCA 1999 Volunteers of the Year. PMID- 10787921 TI - Assisted living meets managed care. PMID- 10787922 TI - HCFA provides training program relief. PMID- 10787923 TI - New rule impacts coverage decisions. PMID- 10787924 TI - Diagnosing and treating dysphagia. PMID- 10787925 TI - A guide to healthy enteral care. PMID- 10787926 TI - Putting food in its place. PMID- 10787927 TI - Nurturing healthy skin. PMID- 10787928 TI - Surveyors focus on nutritional health. PMID- 10787929 TI - Making mealtime enticing. PMID- 10787931 TI - PPS final rule disappointing. PMID- 10787930 TI - Linking nutritional needs for chronic diseases. PMID- 10787932 TI - Bill offers PPS relief for skilled nursing facilities. PMID- 10787933 TI - LTC group opens New Hampshire, Iowa offices. PMID- 10787934 TI - Medicare modernization hits CBO snag. PMID- 10787935 TI - Study finds flaws in PPS design. PMID- 10787937 TI - Beverly settles federal Medicare probe. PMID- 10787936 TI - Shalala supports alternative survey process. PMID- 10787938 TI - JCAHO issues assisted living accreditation principles. PMID- 10787939 TI - Frontier files for bankruptcy, ponders return. PMID- 10787941 TI - Nursing facilities win temporary Y2K protection. PMID- 10787940 TI - GOP bill includes LTC insurance incentives. PMID- 10787942 TI - New rule expands appeals rights. PMID- 10787943 TI - Life at College Park. The challenges and triumphs of the people who live and work in ICFs/MR. PMID- 10787944 TI - It's time to stave off staff stress. PMID- 10787945 TI - Saving Private Ryan's job. PMID- 10787946 TI - SNF financing in a tight credit market. PMID- 10787947 TI - Role of hospice in Alzheimer's care. PMID- 10787948 TI - Health care quality improvement. Resources. PMID- 10787949 TI - Measuring health care quality. How do you know your care has improved? AB - Statistical process control (SPC) was developed in the 1920s as a way of detecting defects in manufacturing processes. During the past decade, SPC has been adopted by service industries and is increasingly being employed by health care organizations. The methodology involves the construction of a control chart to detect variation within processes. Variation is neither good nor bad in itself; therefore, the impact of variation must be interpreted within the context of expected outcomes, acceptable limits, and the process itself. In this article, the concept of statistical process control is explained, and examples are provided to illustrate how SPC can be applied in a clinical setting. PMID- 10787950 TI - Using focus groups to assess primary care patients' satisfaction. AB - The present report describes the use of patient focus groups by a primary health care facility. We review our rationale for using focus groups and the process we used to prepare for and conduct them. We then highlight the results and lessons learned through this experience. Focus groups can be an excellent method for primary care practices to assess the complexities of patient satisfaction issues and engage patients in the continuous quality improvement process. Focus groups can uncover unanticipated issues that surveys fail to identify. Our experience demonstrated that this benefit can be critical in identifying and prioritizing quality of care improvements and that focus group results can be used to make immediate improvements in the quality of care, even though this type of study is not intended to generalize. PMID- 10787951 TI - Quality improvement in health care. Conceptual and historical foundations. AB - During the past decade, quality improvement (QI) has become the primary approach for health care organizations to measure performance and implement change. From a historical perspective, QI has multiple origins: in systems engineering, as a way of defining production processes; in quantitative analysis, as a methodological approach for collecting and analyzing data; and in organizational behavior, as a way of understanding how QI fits with an organization's structure and management philosophy. Although QI and evaluation share similar goals, as modes of inquiry, they developed in very distinct and separate settings, to fulfill different needs. However, these differences are now beginning to merge as QI gains acceptance as a desirable and legitimate approach for health care organizations to enhance service delivery and outcomes. This article describes the conceptual foundations on which QI is based and identifies historical events that have influenced the development of QI and its adaptation by health care organizations. PMID- 10787952 TI - Implementation of the methadone treatment quality assurance system. findings from the feasibility study. AB - This article describes the development and large-scale testing of the Methadone Treatment Quality Assurance System (MTQAS). MTQAS originated as a NIDA (National Institute on Drug Abuse) funded study to assess the feasibility and utility of implementing a performance-based feedback reporting system for narcotic addiction treatment programs. After several years of research to identify a set of valid, reliable, and clinically useful indicators of program performance using patients' in-treatment outcomes, MTQAS was tested in a feasibility study with more than 70 methadone treatment clinics in seven states. For each of nine quarters, participating clinics received a report summarizing their patient outcomes; reports also included summary data permitting comparisons across states, across clinics within a state, and within a clinic over time. In addition, MTQAS included a case-mix adjustment strategy that permitted fair comparisons of performance among programs with systematically different patient caseloads. The structure and operation of MTQAS, selected analyses of the data, and lessons for future performance measurement systems are discussed. PMID- 10787953 TI - Quality improvement in a school health program. Results of a process evaluation. AB - A process evaluation of a school health program designed to improve students' health and educational success is presented. The program included a pediatric nurse practitioner and two nurses placed in three high-risk, urban intervention schools. It is a part of a larger multidimensional pilot intervention project (The Children First Plan). Implemented services and the implementation process are described. Key components for a successful implementation included obtaining buy-in, learning the school culture, defining roles, and keeping a sense of humor. Collaborative efforts among the nursing team, other children-first plan providers, and the school staff improved service delivery. Lack of communication was the primary barrier. In addition, differing philosophies among systems (education, social services, health), problems with referral and feedback, and lack of appropriate providers hampered service delivery. Recommendations for continued program improvement and replication projects are provided. PMID- 10787954 TI - Watch your back (or OSHA will). PMID- 10787955 TI - OSHA bloodborne pathogens standard update. PMID- 10787956 TI - The waiting game: two innovative organizations prove it's not inevitable in healthcare. PMID- 10787957 TI - Emergency department survey: 10% of ED administrators would avoid their own departments. PMID- 10787958 TI - A rapid response reaps rewards: prompt attention to the call bell is the key to patient satisfaction at this hospital. PMID- 10787959 TI - Reducing medication errors. Part I: Fairview health services, Minneapolis. PMID- 10787960 TI - Linking patient learning across a diverse, scattered system. PMID- 10787961 TI - Addressing Africa's agony. Holbrooke and Gore push the U.N. to combat AIDS as a deadly threat to the world's security. PMID- 10787962 TI - The gene machine. To the consternation of his rivals, Craig Venter may have the human DNA 100% mapped by summer. PMID- 10787963 TI - Games species play. What's the secret of life? It may be that our genes are all in the same boat. They sink or swim together. PMID- 10787964 TI - Don't be flued. Those new influenza remedies have great TV commercials, but they won't cure what ails you. PMID- 10787965 TI - Health care: a litmus test. PMID- 10787966 TI - Critical condition. Flu victims are flooding into emergency rooms and discovering another health-care crisis there. PMID- 10787967 TI - Fortified toddlers. PMID- 10787968 TI - How "natural" is rape? PMID- 10787969 TI - Watchdogs who bite. Nutrition activists attack everything from milk to fettuccine Alfredo. Who are these food police? PMID- 10787970 TI - Pros and cons. Hormone therapy can do wonders for women, but a new cancer study underscores the risks. PMID- 10787971 TI - Dirty little diagnosis? A doctor is on trial for calling an illness by a more insurable name, casting light on a common practice. PMID- 10787972 TI - Trouble brewing. A steaming cup is a great comfort, but consider the effects of unfiltered coffee on the heart. PMID- 10787973 TI - When did AIDS begin? PMID- 10787975 TI - Second opinions. When and how should you ask for another physician's advice? Here are some guidelines. PMID- 10787974 TI - The bad and the good. Fresh doubts are cast on a troubled gene-therapy treatment even as the French hint at new advances. PMID- 10787976 TI - Facing a problem status couldn't solve. When my child was diagnosed with severe learning disabilities, we had the fight of a lifetime on our hands. PMID- 10787977 TI - The plague years. As the new century begins, AIDS tightens its medieval death grip on Africa. The anatomy of an epidemic--and its rising human toll. PMID- 10787978 TI - Fighting the disease: what can be done. Curbing the contagion is hard--but not hopeless. A practical prescription. PMID- 10787979 TI - 10 million orphans. For the children who have lost their parents to AIDS, grief is only the beginning of their troubles. The disease's lasting victims. PMID- 10787980 TI - A cause that crosses the color line. AIDS ignores racial and geographic boundaries. But African-Americans can--and should--play a role in battling the scourge. PMID- 10787981 TI - It seems everyone has it. Hospitals and doctors' offices are being overwhelmed by an early and severe outbreak of the flu. PMID- 10787982 TI - An at-home lifesaver? New research looks beyond the Pap smear to find cheaper, simpler tests to detect cervical cancer. PMID- 10787983 TI - Learning to respect a patient's intuitions. A doctor's reflections on how to avoid errors. PMID- 10787984 TI - Getting inside a teen brain. Hormones aren't the only reason adolescents act crazy. Their gray matter differs from children's and adults'. PMID- 10787985 TI - Grace under pressure. Ten years ago, 5,000 people did the exercise routine called Pilates. The number now is 5 million in America alone. But what is it, exactly? PMID- 10787986 TI - A time to decide. Detections of DCIS breast cancer are increasing. So are options for treatment. Which is best? PMID- 10787987 TI - NLM update. Significant increase in hospital library connectivity. PMID- 10787988 TI - Leadership alert: methods for reaching out to administrators, Part II. PMID- 10787989 TI - Balanced Budget Refinement Act restores $2.7 billion to SNFs; industry celebrates victory. PMID- 10787990 TI - BBA revisions include modifications for PPS exempt hospitals. PMID- 10787991 TI - To close or not to close? How to accurately assess the profitability and value of a subacute unit under PPS. PMID- 10787992 TI - Anomaly in Balanced Budget Relief Act could provide strange incentives for treatment of therapy patients. PMID- 10787993 TI - Dust settles around Relief Act; experts examine impact. PMID- 10787994 TI - Key provisions of the Balanced Budget Refinement Act explained in AHCA Q&A document. PMID- 10787995 TI - AMDA white paper helps physicians determine, document medical necessity for SNF patients. PMID- 10787996 TI - Quarterly financial results of post acute/subacute companies. PMID- 10787997 TI - HCFA releases revised enforcement procedures, providers concerned about focus on punishment. PMID- 10787998 TI - HCFA issues first instructions on Balanced Budget Refinement Act. PMID- 10787999 TI - GAO concludes SNF PPS has not decreased overall access to care, but concedes some RUGS-III rates may be too low. PMID- 10788001 TI - HCFA preparing case-mix modifications for release in April; experts to review abt data. PMID- 10788000 TI - The key to balancing costs and care under PPS: get back to the basics. PMID- 10788002 TI - HCFA working on consolidated billing guidance, implementation likely next year. PMID- 10788003 TI - Accurate patient costing under PPS ensures margins, improves efficiency. PMID- 10788004 TI - HCFA updates PPS medical review instructions. PMID- 10788005 TI - How to improve staff retention rates. PMID- 10788006 TI - What does the future hold? Trends in food and nutrition service. PMID- 10788007 TI - Looking ahead... PMID- 10788008 TI - Clinical nutrition staffing needs. Part I: Factors affecting clinical productivity. PMID- 10788010 TI - Improving your listening skills. PMID- 10788009 TI - Cultural competence. Improving knowledge and skills for increasingly diverse clients. PMID- 10788011 TI - Crossing cultural boundaries. PMID- 10788012 TI - Designing regional menus. How the big guys do it. PMID- 10788013 TI - Hospital survey process is evaluated. Two-year assessment by the Office of Inspector General. PMID- 10788014 TI - Clinical nutrition staffing. Part II: Measuring patient acuity. PMID- 10788015 TI - Women's health care and cardiovascular disease management. PMID- 10788016 TI - The most common killer of women is not the most feared. PMID- 10788017 TI - Roundtable discussion ... cardiovascular disease management programs. PMID- 10788018 TI - The role of women in health care decision making. PMID- 10788019 TI - Roundtable discussion ... are MCOs implementing programs to meet the challenges of baby-boomer women entering menopause? PMID- 10788020 TI - Prevention in the Medicare care environment. PMID- 10788021 TI - The epidemiology of cardiovascular disease in women. PMID- 10788022 TI - How satisfied are mothers with 1-day hospital stays for routine delivery? AB - CONTEXT: Payers and health plans are encouraging shorter hospital stays after routine vaginal delivery. OBJECTIVE: To assess the satisfaction of mothers who had 1-day or 2-day stays after routine delivery. DESIGN: We mailed questionnaires to mothers 7 to 9 months after delivery. The self-administered survey contained questions about the mothers' satisfaction with the care they received, clinical complications, and the mothers' preparedness after discharge. SETTING: A mixed staff, network-model managed care plan in Minnesota that encourages but does not require 1-day hospital stays after routine delivery. PARTICIPANTS: All plan members who delivered a baby vaginally in the first quarter of 1995 (n = 1009). RESULTS: 56% of the mothers responded to the survey. Of these, 202 had 1-day stays and 292 had 2-day stays. Mothers with 1-day stays were more likely than mothers with 2-day stays to report that their length of stay was "too short" (75% vs. 37%; P < 0.001), and 81% of mothers with 1-day stays would want to stay longer if they had another child. The frequency of self-reported maternal or infant complications did not differ substantially between the two groups. More mothers with 1-day stays than mothers with 2-day stays received home health care visits (44% vs. 10%; P < 0.001). CONCLUSION: Although length of stay does not seem to be related to clinical outcomes after vaginal delivery, mothers with 1 day stays are less satisfied with their length of stay. PMID- 10788023 TI - A comparison of resource utilization in nurse practitioners and physicians. AB - CONTEXT: Nurse practitioners increasingly provide primary care in a variety of settings. Little is known about how resource utilization for patients assigned to nurse practitioners compares with that for patients assigned to physicians. OBJECTIVE: To compare health care resource utilization for adult patients assigned to a nurse practitioner with that for patients assigned to a resident or attending physician. DESIGN: Prospective, quasi-randomized study. SETTING: Primary care clinic at a Veterans Affairs medical center. PATIENTS: 450 new primary care patients: 150 were assigned to a nurse practitioner, 150 to a resident physician, and 150 to an attending physician. OUTCOME MEASURES: We collected data on laboratory and radiologic testing, specialty care, primary care, emergency or walk-in visits, and hospitalizations over a 1-year period. We also collected information on baseline chronic illnesses, blood pressure, and weight. RESULTS: Resource utilization for patients assigned to a nurse practitioner was higher than that for patients assigned to a resident in 14 of 17 utilization measures (3 were statistically significant) and higher in 10 of 17 measures when compared with patients assigned to an attending physician (3 were statistically significant). None of the utilization measures for patients in the nurse practitioner group was significantly lower than those for either physician group. CONCLUSIONS: In a primary care setting, nurse practitioners may utilize more health care resources than physicians. PMID- 10788024 TI - Can percent free prostate-specific antigen reduce the need for prostate biopsy? AB - BACKGROUND: In a recent multicenter study, percent free prostate-specific antigen (PSA) enhanced the specificity of PSA testing in prostate cancer screening. OBJECTIVE: To determine whether the percent free PSA could be as effective in reducing the need for biopsy in a managed care setting as in an academic setting. SETTING: Kaiser Permanente Northwest Region (Portland, Oregon) and Kaiser Permanente Oakland/Berkeley (California). DESIGN: Prospective blinded study conducted by using Hybritech Tandem PSA and Hybritech Tandem free PSA assays (Beckman Coulter, Inc., Fullerton, California). PARTICIPANTS: 250 men (63 with prostate cancer and 187 with benign prostate conditions) who were older than 40 years of age, had a PSA level of 4.0 to 10.0 ng/mL, and had a histologically confirmed diagnosis. MAIN OUTCOME MEASURES: Sensitivity and specificity of percent free PSA. RESULTS: The median percent free PSA values for patients with cancer (free PSA, 13%) significantly differed from that for patients without cancer (free PSA, 17%) (P = 0.001). When a free PSA cutoff of 25% was used, the sensitivity was 97% (95% CI, 92% to 100%) and the specificity was 13% (CI, 8% to 18%). These results were not significantly different from those obtained in the multicenter study (95% sensitivity, 20% specificity for a free PSA cutoff of 25%). CONCLUSION: The results obtained in a managed care organization were similar to those obtained at large university medical centers and show that the percent free PSA can be used to enhance the specificity of PSA testing for prostate cancer. PMID- 10788025 TI - Adolescent health care visits: opportunities for brief prevention messages. AB - CONTEXT: It has been suggested that clinicians should increase efforts to modify and prevent risky behavior in adolescents. Professional organizations have proposed recommendations about access to care and preventive services, but it is difficult to know where and how to most effectively deliver such services. PRACTICE PATTERN EXAMINED: Clinic visits among adolescent HMO members (14 to 17 years of age). SETTING: Kaiser Permanente Northwest Division, a medium-sized, nonprofit, group-model HMO in the Pacific Northwest. DATA SOURCES: Two administrative databases (one for membership and one for outpatient utilization). RESULTS: A total of 22,626 adolescents who met the inclusion criteria were identified. Of these, 62% (more than 14,000 adolescents) were seen in a primary care clinic within 1 year; almost 83% (more than 18,000 adolescents) were seen within 2 years. There were several opportunities for follow-up for adolescents who had at least one visit in 1995: 60% had more than one visit during 1995, and 80% had more than one visit over the 2-year span of 1995 and 1996. The largest number of adolescent visits occurred in August through November, and most visits took place in the afternoon. CONCLUSIONS: Primary care visits in an HMO present an excellent opportunity to reach many teenagers outside of a school setting. Short-term educational or prevention programs would be optimal during late summer and fall; additional staff members may be able to present these programs after school once school begins. PMID- 10788026 TI - Potential benefits of regionalizing major surgery in Medicare patients. AB - CONTEXT: Given the strong "volume-outcome" relations observed with many surgical procedures, concentrating surgery in high-volume hospitals could substantially reduce the number of surgical deaths. We explored the potential benefits of regionalizing 10 high-risk procedures for the 38 million Americans enrolled in Medicare. COUNT: Number of lives saved in 1 year. CALCULATION: Current number of deaths occurring with each procedure multiplied by the average mortality reductions that plausibly could be achieved with regionalization. DATA SOURCE: The current number of surgical deaths was obtained from the 1995 MEDPAR file of the Medicare claims database. Expected mortality rate reductions with regionalization, estimated from published volume-outcome studies, were tested over a wide range in sensitivity analysis. RESULTS: Of 381,000 Medicare patients undergoing any 1 of the 10 procedures in 1995, approximately 17,000 surgical deaths occurred. The total number of lives saved by regionalization depends on assumptions about the mortality reductions likely to be achieved, varying from 853 (5% reduction) to 4266 (25% reduction). Regionalizing common, intermediate risk procedures (e.g., cardiovascular procedures) would save far more lives than regionalizing less-common, higher-risk operations (e.g., major cancer resections). CONCLUSIONS: Even with conservative assumptions about reduction in surgical mortality likely to be achieved, the benefits of regionalizing major procedures in Medicare patients could be substantial. Policymakers should focus on common procedures before less-common, high-risk operations. PMID- 10788027 TI - Performance measures: the destination or the journey? PMID- 10788028 TI - Physician autonomy, patient choice, and immunization performance measures. PMID- 10788030 TI - Who cares what surgeons think? PMID- 10788029 TI - Performance measures for ethics quality. PMID- 10788031 TI - Radiologic tests after a new diagnosis of breast cancer. AB - CONTEXT: Radiologic tests may be overused in the staging of newly diagnosed breast cancer. OBJECTIVE: To determine the frequency with which radiologic tests are used in women with newly diagnosed breast cancer and the yield of such tests in these patients. METHODS: We used the tumor registry database from the Hoag Cancer Center, Newport Beach, California, to identify and classify the disease stage of all patients with breast cancer who received a diagnosis from or were initially treated by Hoag staff from 1990 to 1994. After excluding patients with unknown tumor (T) and lymph node (N) status, we retrospectively determined the frequency with which radiologic tests were performed within 4 weeks of diagnosis and the proportion of these tests that detected metastatic disease. RESULTS: A total of 1910 radiologic tests, including 646 bone scans, 637 chest radiographs, and 627 other tests, were obtained in 1167 patients with a known TN status. Radiologic tests were performed in 42% of patients with carcinoma in situ, but none of the 183 tests detected metastases. Eight hundred twenty-eight radiologic tests were performed in patients who were classified as having stage I disease on the basis of TN criteria. Only three of these tests (0.4%) detected metastatic disease, and all three were performed in one patient with bone pain. For patients who were classified as having stage IIA, stage IIB, or stage III disease on the basis of TN criteria, 5 of 410 tests (1.2%), 20 of 294 tests (6.8%), and 33 of 195 tests (17%), respectively, yielded positive results. CONCLUSIONS: Radiologic staging tests are overused in patients with newly diagnosed, early-stage breast cancer. These tests are unnecessary in patients with breast cancer who have 1) a tumor that is 5 cm in diameter or smaller, 2) no axillary lymphadenopathy on physical examination, 3) normal results on blood chemistry tests, and 4) no symptoms or physical findings of metastatic disease. PMID- 10788032 TI - Improving general practice care of patients with asthma or chronic obstructive pulmonary disease: evaluation of a quality system. AB - CONTEXT: During the past decade, several guidelines on the management of chronic obstructive pulmonary disease and asthma have been developed. However, strategies for implementing these guidelines have not been systematically evaluated. OBJECTIVE: To test a quality system intended to improve general practitioners' compliance with recently established guidelines. DESIGN: Before-after study with concurrent controls. UNIT OF ANALYSIS: 19 general medical practices in the Netherlands (14 intervention practices and 5 control practices). INTERVENTION: A quality system with five components: identification of barriers, documentation, education, feedback, and peer review. PATIENTS: Outpatients 16 to 70 years of age with asthma or chronic obstructive pulmonary disease. MEASUREMENTS: The number of consultations for respiratory symptom monitoring, measurement of peak expiratory flow rate, prescription of anti-inflammatory agents, monitoring of medication compliance and inhalation technique, and influenza vaccination. RESULTS: The percentage of patients who had two or more consultations per year increased significantly in the intervention practices (median, 27% of patients before the intervention vs 82% of patients after the intervention; P < 0.01), as did the percentage of patients who had at least one measurement of peak expiratory flow rate (median, 10% of patients before the intervention vs 84% of patients after the intervention; P < 0.01). The percentage of patients who received a prescription for anti-inflammatory agents did not increase significantly. No significant changes were seen in the control practices. Physicians in the intervention practices were more likely to monitor medication compliance and inhalation technique. No difference was found in frequency of influenza vaccination. CONCLUSION: The quality system improved guideline compliance in some areas but not in others. PMID- 10788033 TI - Alternative explanations for poor report card performance. AB - CONTEXT: Many managed care organizations grade physician groups with "report cards" developed from administrative data sets and chart reviews. OBJECTIVE: To investigate the accuracy of five report cards on a single group practice. DESIGN: Determination of report card accuracy by using the practice capitation list and a review of the patients' medical records. SETTING: Academic practice in Philadelphia, Pennsylvania (19 physicians), evaluated with five report cards by two capitated health plans between 1994 and 1997. RESULTS: Four major problems were uncovered. First, four of the five report cards included patients who were enrolled in our practice for only a portion of the reporting year (for the four report cards, the proportion of partial-year enrollees was 8%, 15%, 23%, and 100%). Second, there was a considerable number of false-positive diagnoses in the administrative algorithms. Eight of the 61 patients labeled with hypertension did not have this condition (error rate, 14%). Other error rates were 44% for coronary artery disease, 50% for congestive heart failure, 33% for atrial fibrillation, and 0% for diabetes. Third, the administrative data often failed to capture laboratory data. Laboratory performance measures for patients with diabetes (hemoglobin A1c and cholesterol measurement and screening for microalbuminuria) were 3 to 10 times higher when assessed by chart review. Finally, the uniformly small sample sizes used in the report cards make the estimates of performance imprecise. No report card reported 95% CIs. CONCLUSION: Five report cards on a group practice contained methodologic problems that led to systematic underestimation of the practice's performance. Larger surveys are needed to determine the accuracy of report cards in current use. PMID- 10788034 TI - Medication review and documentation in physician office practice. AB - CONTEXT: Adverse drug reactions and drug-drug interactions are common. Medication induced morbidity might be prevented through the documentation of medicines in the medical record and review of the medical record before new medications are prescribed. PRACTICE PATTERN EXAMINED: Documentation and review by primary care physicians of patient use of prescription drugs, over-the-counter drugs (OTCs), nutritional supplements, and herbal and other alternative treatments. DATA SOURCE: A stratified random sample of 1802 internists and family practitioners from the American Medical Association Physician Masterfile was surveyed; 655 physicians responded (response rate, 36%). RESULTS: 99.8% of physicians reported documenting prescription drugs in the medical record. Fewer reported documenting OTCs (68%) or nutritional supplements (63%); only 47% documented herbal and other alternative treatments. Almost all respondents reported reviewing prescription medications before prescribing a new therapy (99.8%), but only 86% reported reviewing OTCs at the same time. Fewer than half of physicians reported reviewing nutritional supplements or herbal and other alternative treatments before prescribing a new therapy. CONCLUSIONS: This study draws on self-reported data, and the response rate was low. Thus, the results probably overestimate actual rates of documentation and review. Review and documentation of nonprescription substances are uncommon in primary care practice. PMID- 10788035 TI - Is hospitalism new? An analysis of medicare data from Washington State in 1994. AB - CONTEXT: Managed care, increased disease severity, and more complex treatment options may be reasons for the recent enthusiasm for "hospitalists"--physicians who specialize in the care of inpatients. It is not clear, however, whether hospitalism is a new model for caring for inpatients or merely a new description for previously existing practice patterns. PRACTICE PATTERNS EXAMINED: The proportion of physician visits occurring in the hospital before the introduction of the term hospitalists. Five specialties were examined: family/general practice, general internal medicine, cardiology, gastroenterology, and pulmonology. DATA SOURCE: 1994 Medicare Part B claims data for beneficiaries 65 years of age and older who received all of their care in Washington State. RESULTS: For the average family/general practitioner, 10% of all Medicare visits occurred in the hospital. Corresponding figures for the other specialties were 20% for general internists, 36% for cardiologists, 38% for gastroenterologists, and 45% for pulmonologists. A substantial number of physicians devoted most of their Medicare effort to inpatient care (i.e., hospital visits > 50% of total visits). If this definition were used as a proxy for hospitalism, 4% of family/general practitioners, 10% of general internists, 20% of gastroenterologists, 29% of cardiologists, and 37% of pulmonologists would have been considered hospitalists in Washington State during 1994. On the other hand, 35% of family/general practitioners, 18% of general internists, 7% of both gastroenterologists and pulmonologists, and 4% of cardiologists did not bill Medicare for any inpatient visits and could reasonably be categorized as "officists." CONCLUSION: Physicians vary considerably in the proportion of their workload that occurs in the hospital or outpatient setting. Even before the term was coined, a considerable number of physicians were de facto "hospitalists." PMID- 10788036 TI - Improving quality improvement research. PMID- 10788037 TI - Risky business. PMID- 10788039 TI - An orthopedist questions the utility of surgery for back pain. PMID- 10788038 TI - Informed consent for screening by community sampling. PMID- 10788040 TI - How patients with diabetes perceive their risk for major complications. AB - CONTEXT: To educate patients with diabetes about their illness and to motivate these patients to pursue intensive treatment, physicians often inform them about their risk for serious complications. However, little is known about patient perceptions of these risks. OBJECTIVE: To compare patient perceptions of risk for major complications of diabetes with actual risk for these complications. DESIGN: Structured interviews were done to obtain the patient's estimate of their risk for complications. To generate estimates of actual risk for each patient, we used a simulation model based on the Diabetes Control and Complications Trial (DCCT). SETTING: Four university-affiliated diabetes clinics in the midwestern United States. PATIENTS: 139 patients with type 1 diabetes mellitus. MAIN OUTCOME MEASURES: Probability of blindness, end-stage renal disease (ESRD), and lower-leg amputation over 20 years. RESULTS: Participants were young (mean age, 30 years) and reported having had diabetes for an average of 15 years. Seventy-nine percent reported their current diabetic therapy to be "intensive." Ninety-eight percent had completed high school, and 51% were college graduates. The patients' estimates of their risks far exceeded the DCCT estimates for all three complications. The mean patient estimate of the risk for blindness was 31.6% (DCCT estimate, 17.0%), of the risk for ESRD was 33.7% (DCCT estimate, 8.7%), and of the risk for amputation was 25.1% (DCCT estimate, 1.9%). Similarly, patients overestimated the benefit of intensive therapy. They estimated, on average, that intensive therapy would result in a 17.0% absolute risk reduction for blindness (DCCT estimate, 12.2%), an 18.1% risk reduction for ESRD (DCCT estimate, 7.0%), and a 14.2% risk reduction for amputation (DCCT estimate, 1.2%). CONCLUSION: Patients with diabetes overestimated their risk for major complications and the benefits of intensive treatment. PMID- 10788041 TI - False claims act: theories of liability and constitutionality. PMID- 10788042 TI - Expansion of use of breach of confidence: addressing unauthorized disclosure of patient information. PMID- 10788043 TI - Disabling the pacemaker: the heart-rending decision every competent patient has a right to make. PMID- 10788044 TI - Why these biotechs are as hot as net stocks. Wall Street's betting big on a genetic technology that burned it badly before. Monoclonal antibodies still don't cure cancer--but this time they may actually pay off. PMID- 10788045 TI - Blessings from the book of life. Decoding the human genome will yield a bounty of biotech miracles that will transform our lives in the next 40 years. PMID- 10788046 TI - Biotech's wonder ride. PMID- 10788047 TI - Hospitals should look at radiology as profit center. PMID- 10788048 TI - Tenet, Columbia/HCA invest in online procurement companies. PMID- 10788049 TI - The ultimate quality indicator. PMID- 10788050 TI - When there's so much technology available, how does a hospital decide what to buy? PMID- 10788051 TI - Taking care of the hospital's caregivers. PMID- 10788052 TI - What's your flu season strategy for 2001? PMID- 10788053 TI - Hospitals 'too slow' to read deteriorating finances, new EY 'crisis' partner says. PMID- 10788054 TI - Despite doing things right, Mercy Hospital starves from poor reimbursements. PMID- 10788055 TI - Only 13 hospitals make the 'top hospital' lists of both NRC, HCIA. PMID- 10788056 TI - Telemedicine will grow 40% annually, help health systems create 'centers of excellence'. PMID- 10788057 TI - Hospitals need to act to reduce bad debt writeoff, which exceeds profit by 3-1. PMID- 10788058 TI - More small group practices, fewer large group practices have capitation revenue. PMID- 10788059 TI - Hospital, home health, assisted living stocks down, e-commerce, lab stocks up in 1999. PMID- 10788060 TI - They're b-a-a-c-k: scrub suit popularity and costly employee theft. PMID- 10788061 TI - Rural hospital ups security to deal with current and potential problems. PMID- 10788062 TI - Study: incidents in parking areas leading source of liability claims. PMID- 10788064 TI - Salary surveys: income of healthcare security directors up 3.1 to 8.5%. PMID- 10788063 TI - Plane crashes into neighborhood: how hospital emergency plan worked. PMID- 10788065 TI - New children's hospitals: anticipating and revising security and safety approaches. PMID- 10788066 TI - Hospital security officer wins award as security 'pacesetter'. PMID- 10788067 TI - Mobilizing toward regional accreditation. AB - In January 1999, the Eastern Regional Health Board began planning for a Regional Accreditation scheduled in November 1999. This would be the first time that our two programs (Addiction Services and Public Health), and two of the nine acute care facilities in our Region would be accredited. This article describes a regional accreditation planning process. It suggests the resources required for such a project, suggests a workable time line, and indicates a communication strategy. Mobilizing over 1000 staff, physicians, and the communities toward Regional accreditation was a challenge for the Steering Team. Defining a plan of action with defined time lines proved helpful; defining team chairs who were flexible and positive was vital, and engaging the communities in our review was rewarding. Informing the public was completed using strategies borrowed from our Newfoundland counterparts (an accreditation mural and a regular newsletter), and our own creative strategies such as "Sharing Day" and engaging community health board involvement on accreditation teams. The project development was initiated by a steering committee, and spanned 21 teams. Both a communication and education plan were important in keeping the board, staff, physicians and residents of the Region informed. PMID- 10788068 TI - Resource allocation in integrated delivery systems and healthcare networks: a proposed framework to guide ethical thinking. AB - Drawing on a management perspective and the literature, this article suggests an ethical framework to be used at the meso or community level of resource allocation in a Canadian setting. The suggested framework enlarges on the program level framework developed by Meslin et al primarily by building in stakeholder inclusiveness and public accountability, both of which are essential to resource allocation at the population-based level. PMID- 10788069 TI - Integrated health information systems based on the RAI/MDS series of instruments. AB - There is a growing need for an integrated health information system to be used in community, institutional and hospital based settings. For example, changes in the structure, process and venues of service delivery mean that individuals with similar needs may be cared for in a variety of different settings. Moreover, as people make transitions from one sector of the healthcare system to another, there is a need for comparable information to ensure continuity of care and reduced assessment burden. The RAI/MDS series of assessment instruments comprise an integrated health information system because they have consistent terminology, common core items, and a common conceptual basis in a clinical approach that emphasizes the identification of functional problems. PMID- 10788070 TI - Measuring patient satisfaction--be prepared. AB - This article describes the approach used by the Queen Elizabeth II Health Sciences Centre to measure inpatient satisfaction at the recently merged institution. It presents some of the issues that arose in the planning stages, including use of prior consent. It is important to plan the methodology, the execution, the communication strategy and the follow-up before the process begins to ensure the results are credible, communicated and translated into quality improvement actions. PMID- 10788071 TI - Introducing "evidence" into board decision-making. AB - The purpose of this article is to report on the results of a workshop that introduced evidence-based decision-making techniques to Board members of regional health authorities in Alberta. results and conclusions: The workshop demonstrated that it is possible to design a process for the incorporation of evidence in administrative decision-making. The participants demonstrated that they were able to apply scientific evidence in administrative decision making and that the decisions taken were reasonably consistent. Also, in the absence of evidence, values took precedence in the decision-making process and the decisions taken were less consistent. PMID- 10788072 TI - Results of implementation of a contemporary model for ambulance diversions in an integrated healthcare delivery system. AB - This brief report contains a description of a contemporary, coordinated new system for ambulance diversions, which was implemented in Edmonton area's Capital Health Region in January 1999. The development of this new system was precipitated by the combination of increasing pressures within the acute care system especially being felt within our emergency departments, and mounting evidence that the existing system for ambulance diversions was ineffectual in providing temporary relief of these pressures. The nature of the previous and the new system are compared and contrasted, and data are included to demonstrate that the new system has led to marked reductions in the number of ambulance diversions experienced regionally. PMID- 10788073 TI - Evaluation of communication surrounding the implementation of the resident assessment instrument. AB - Two outcomes of the introduction of the Resident Assessment Instrument at Sunnybrook Health Sciences Centre in 1996 were (1) a communication plan and (2) patient unit RAI communication centres. This article describes a survey that was used to evaluate the effectiveness of the plan and centres in implementing the RAI Staff identified the InfoRAInbow Newsletter as the most useful communication strategy. Satisfaction with communication about the RAI implementation was only 57 percent. Further investigation of this level of satisfaction is required. This result may be attributed to the "choices" staff make as to whether or not to read information, or to environmental factors that occurred at the time of the survey's completion. Future directions for improving the RAI Communication Plan should focus on environmental factors that may impede communication of information, especially information that affects integration of the RAI into the clinical care system. PMID- 10788074 TI - Navigating ambulatory referral: a standardized process. AB - Significant funding and structure changes to healthcare in Ontario in the mid 90's led The Hospital for Sick Children in Toronto to examine patient referral processes. In an effort to streamline access and encourage more appropriate referrals, the hospital tested and implemented three major changes. This article outlines these changes using the PDSA (Plan, Do, Study, Act) improvement framework and summarizes the results from this project. PMID- 10788075 TI - Is there room for managed care in Canada? A preliminary inquiry of physicians. PMID- 10788076 TI - Community-based care. Interview by Matthew D. Pavelich. PMID- 10788077 TI - Online solution for tracking payer data. AB - Keeping on top of constantly changing managed care policy requirements can be labor intensive and prone to error. At the University of Missouri Health Sciences Center, over 20 percent of patients are under managed care arrangements for services. The center maintains over 50 contracts with insurers. The medical center thus faces the logistical nightmare of keeping information regarding insurer contact information, pre-certification and prior-authorization policies up-to-date and accessible to front-line clerical and nursing support staff. The medical center is trying to solve the problem by keeping real-time information stored on an easily accessible, searchable Web site. PMID- 10788078 TI - Clouds and rainbows on the HIPAA horizon. AB - In the planning phase for several years, the administrative simplification provisions of the Health Insurance Portability and Accountability Act of 1996 (HIPAA) finally appear ready for implementation. The Health Care Financing Administration (HCFA) expects that the major HIPAA measures will be released in late 1999. For group practices, it is expected that implementation of these provisions will be complicated and consume significant amounts of time and money. Still, the end result should justify the effort. PMID- 10788079 TI - Got voice? AB - Voice recognition technology has come a long way since its start and has several applications in modern health care practice, especially in the development of electronic medical records. However, interested group practice managers need to look past the hype to get a realistic picture of the advantages and disadvantages of the technology in its various forms, as described in this article. PMID- 10788080 TI - Care management and technology enhance care delivery. AB - Many health care delivery organizations have experienced a flattening in the rate of their ability to decrease health care costs. Providers--if they are willing to take responsibility for care management and make appropriate investments to support the initiatives--are the logical party to take responsibility for improving care and cost control. Taking responsibility for care management, however, requires the tactical deployment of information technology to enhance care management. This article outlines what is required for information technology to improve care and cut costs. PMID- 10788081 TI - Trends and tips for online recruiting and job hunts. AB - The Internet is providing new ways to find a new job or a great job candidate. Online job banks can make searches easy, but it's best to have a sense of what type of job bank will best serve your purposes. This article describes six different business models employed by Internet job banks and outlines their advantages and disadvantages. A list of online resources is included. PMID- 10788082 TI - Selecting a clinical system. AB - For more than three decades, health care information systems have built upon the administrative and financial information needs of the health care delivery organization. Industry analysts project that administrative and financial information systems will continue to be important to health care, but post-Y2K resources increasingly will fall on the purchase, installation and support of clinical documentation and reporting tools. This manuscript provides a brief overview of the electronic medical record (EMR), and identifies and explains the need for EMRs in health care. This article also provides strategies for medical group managers and information systems managers to successfully select and negotiate a contract for an electronic medical record. PMID- 10788083 TI - Physician-patient electronic communications. AB - Patients who do not have adequate access to their physicians may feel isolated from decisions concerning their medical treatment and care and believe they have been abandoned to an impersonal health care system. Some physicians are addressing these concerns by adopting new technologies such as e-mail communication. The authors discuss some of the promises and challenges unique to physician-patient electronic communication. PMID- 10788084 TI - Where does IT fit in your practice? Information technology trends. AB - The future role of information technology in the medical industry is never clear, but we can identify some trends, such as electronic medical records and database management systems. Many of these trends can boost productivity for practices, but all managers need to carefully analyze their needs and understand the products to see how they may or may not help their individual practice. PMID- 10788085 TI - New safe harbors to federal antikickback law issued. PMID- 10788086 TI - Past due patient balances can be reduced with proper planning. PMID- 10788087 TI - Prevention key to meeting OSHA ergonomics standard. PMID- 10788088 TI - 2000 and beyond: are you ready for the new generation of risk adjusters? PMID- 10788089 TI - Medicare benefits extended for immunosuppressive drugs. PMID- 10788090 TI - Internal controls help prevent financial misconduct. PMID- 10788091 TI - Satisfying values--yours and theirs: a talk with Manny Elkind. Interview by Richard L.Reece. AB - Richard Reece, MD, interviewed Mansfield (Manny) Elkind on September 22, 1999 to talk about satisfying values and flexible thinking as keys to influencing physicians. Manny discusses why it's difficult for people to create lasting change in the workplace without recreating themselves. The effort to influence others often focuses on getting disciplined about changing behaviors, with the hope that feelings and attitudes will also change. This approach rarely works because behavior is changed only when values or beliefs change. People will get passionate and committed to achieving the organization's goals when they believe that their values will be satisfied as well. The leader's responsibility is to find ways of satisfying people's values, in addition to convincing them to use the organization's values as guides for behavior. A process is explored that helps people discover each other's values and preferences and transforms resistance to commitment. PMID- 10788092 TI - Paving the road to hell: why change programs ultimately fail. AB - A successful change program must have clearly delineated and honestly defined goals. Change requires conviction, money, and effort and the willingness to invest all three. Change programs fail when any of these requirements are lacking. Many health care executives have the conviction, but are unwilling to spend the money or expend the effort. Some lack conviction, despite having money; the effort is merely spinning wheels with no forward motion. Often all three are lacking, despite an outward appearance to the contrary--the result of management delusions. Employees are quick to see through misleading and insincere schemes. It is much easier to foster trust from the outset than to rebuild it after it has been violated. PMID- 10788093 TI - Old myths die hard. PMID- 10788094 TI - Accelerating acceptance. AB - Innovations are either accepted or rejected in large part because of their implementation--sometimes without regard to improvements over existing techniques. By understanding the dynamics of how innovations are adopted and by whom, physician executives will have insight into influencing others. All adopters are not created equal. They fall into five categories, beginning with those who embrace innovation, even seek it out. In any given organization, or the population in general, there are: (1) innovators (2.5 percent); (2) early adopters (13.5 percent); (3) early majority (34 percent); (4) late majority (34 percent); and (5) laggards (16 percent). As these categories imply, each group has specific personalities related to adopting innovations that can be identified and used to implement new policies and procedures. By identifying and encouraging innovators and early adopters to think outside the box and then gaining critical mass through the early majority, who also act as opinion leaders, organizations can accelerate the pace of adopting innovations. New processes are adopted when opinion leaders initiate new practices, whose results can be tangibly discerned by the majority of adopters. PMID- 10788095 TI - Physician executives influencing other physicians. PMID- 10788096 TI - From resistance to attraction: a different approach to change. AB - How can physician executives interested in promoting change be more effective at the task? This article explores how to create an attraction towards change, as opposed to viewing change as overcoming resistance. Learning to recognize naturally occurring change, identify attractors, explore the rationality of others' points of view, and reduce risk are clear and constructive insights from research and emerging systems science. Other ideas to consider are: Understand the issues of those you wish to change, create changes that are "exothermic," produce system changes, accept responsibility, learn from failed efforts, and focus on building relationships of trust. PMID- 10788097 TI - Creating attractors for improving access to clinical offices. PMID- 10788098 TI - Creating attraction through prototyping. PMID- 10788099 TI - The fulcrum and the lever. PMID- 10788100 TI - Benchmarking: the key to influencing physicians. AB - Managing physicians to achieve cost reductions can seem impossible, especially when managed care penetration is low. Physicians feel little pressure to change when asked merely to cut costs, especially when their boat is not rocking. But physicians will respond to benchmarking data on CPT-coded procedures that are directly comparable to their own practices. When surgeons see that others take less time to perform a procedure and/or use fewer and lower cost supplies, their competitive spirits are aroused. They become inquisitive about why this is so and then are eager to change by trying new methods and improving their techniques. Science is the key motivator, not savings. When benchmarking recommendations are implemented in a facility, better practice and substantial cost savings are the positive results. PMID- 10788101 TI - Pre-certification: lessons from diagnostic imaging. AB - Recently, managed care has received a great deal of criticism from the public and press, some of it richly deserved. One of the areas of greatest concern is pre certification requirements and possible denial of services. Experience with programs of this sort in diagnostic imaging suggests that, implemented correctly, the consultative interface created by a pre-certification program can be of tremendous value in physician education. Appropriate behavior modification programs need to be crafted with an educational rather than economic bias and the methodology not be totally abandoned simply because it has been misused by some. The pre-certification process can be used as a vehicle to disseminate information about rapidly emerging new therapies and technologies. PMID- 10788102 TI - Improving performance in a contracted physician network. AB - Health care organizations face significant performance challenges. Achieving desired results requires the highest level of partnership with independent physicians. Tufts Health Plan invited medical directors of its affiliated groups to participate in a leadership development process to improve clinical, service, and business performance. The design included performance review, gap analysis, priority setting, improvement work plans, and defining the optimum practice culture. Medical directors practiced core leadership capabilities, including building a shared context, getting physician buy-in, and managing outliers. The peer learning environment has been sustained in redesigned medical directors' meetings. There has been significant performance improvement in several practices and enhanced relations between the health plan and medical directors. PMID- 10788103 TI - Watching the falling barometer. AB - How can physician executives create a vision, a strategy, in the face of such overwhelming forces for change? The answer has two pieces. The first is the Weather Channel: scanning the future for warning, for opportunities, for new business possibilities. The second leads us to such questions as: What is your situation? Financially? In market terms? It leads us, as well, back to the question: For you and your institution, what is your reason for being in this business? In other words, what is your foundation? If you can become clear about who you are and what you are here for in the long run, and match that with some sense of the technologies and the political and financial pressures headed your way, then you can begin to create a vision of a future that works for you. In the coming years, we will begin to create entire new ways of doing health care, new roles for hospitals, new types of medicine--and the time to begin the creation is now. If you wait until the hurricane hits, it will be too late. PMID- 10788104 TI - Are you obsolete? AB - Are you vulnerable, regardless of length of service at your organization and your unique skill sets? There are ways to test vulnerability and assemble some hard evidence that your management role makes a difference. You need to conduct a self test for obsolescence. Ask yourself the following questions: Are your skills state-of-the-art? As a manager, how do you compare with others doing the same, or similar, job at competing organizations? Is your role essential? Where does your job fall into the big picture? Can you be replaced easily? If a thorough examination of your skills and your role convinces you that your contribution returns more to the organization than your salary, can you prove it? Consider these strategies: (1) Put together a portfolio, (2) ensure your boss' support, (3) advertise your successes, and (4) cultivate recruiters. The best reason to analyze your value to the organization is that if you are laid off, getting another comparable job--or a better one--will be far less of a hassle. PMID- 10788105 TI - The grief involved in change. AB - In this era of rapid change, people need to be helped through the grieving process at work. "By acknowledging and articulating what is involved, you facilitate people's movement through it ... successful people in growing organizations need to learn to move through the process as quickly and efficiently as possible and help others do the same." Morris Shectman, in Working Without a Net says, "Contrary to the old paradigm--which held that others don't have a right to know about your personal life--the new paradigm says that it's a necessity that they know." If people are to move through the grief that is caused by undesired change, they will have to tell some of their personal feelings. "Each stage of the process--shock and denial, anger, bargaining, depression, and acceptance--is functional ... when people go through the process in a healthy manner, they'll recycle through it in a diluted fashion." Leaders in the organization can help people start the process or move through it if they get stuck at a particular stage. PMID- 10788106 TI - Real executives make career moves. Will you? AB - Not all physician executives have accepted the career move as a key part of executive life. They prefer to stay put and they often do just that. While clinicians may have the luxury of choosing a single geographic location and remaining there through retirement, physician executives often have to reorder their priorities to give the position greatest significance in career decision making. Spouses and families need to be educated to the new reality of executive life--sometimes, to support an important career opportunity, a move is required. Physician executives unwilling to make career moves limit their career potential. PMID- 10788107 TI - Graduate medical education funding crisis. AB - Medicare, the major funding source for graduate medical education, is under intense scrutiny. Some think that Medicare should not continue to overtly pay to train graduate physicians, while others believe continued direct funding is an important social obligation. Recent policy discussions could dramatically transform the way graduate physician training is funded and teaching hospitals are paid. This is an important debate because of the strong influence Medicare has in determining the willingness of other payers to contribute to graduate medical education. Funding reductions are already threatening the viability of the U.S. teaching hospital system. PMID- 10788108 TI - The levers of influence. AB - Physicians often determine the demand for health care services, as well as control the clinical processes aimed at improving health outcomes at the individual and population level. Given their important role in enhancing health status and improving the health care delivery system, it is critical that physician executives master the tools necessary to positively influence physician behavior. But changing behavior is far more complex than "doing it or not doing it." The Nike slogan "just do it" is motivating, but over-simplified. The roots of human change include: consciousness-raising, emotional arousal, commitment, helping relationships, self-reevaluation, reward, and environmental control. A model to effectively influence behavior is presented and includes setting clear expectations, measuring and monitoring performance, providing feedback, and rewarding and recognizing improvement. If all else fails, try discipline. This five-step approach is based on the science of human behavior and working with physicians in diverse settings, ranging from academic medical centers to small practices. PMID- 10788109 TI - A new industrial order for physicians: a talk with Jeff C. Goldsmith, PhD. Interview by Richard L. Reece. AB - Richard L. Reece, MD, interviewed Jeff C. Goldsmith, PhD, President of Health Futures, Inc. on October 12, 1999 to discuss how the Internet will affect health care delivery in the millennium. One of the most profound changes that he sees is how the relationship between physicians and patients will be altered. Empowered consumers are where the real revolution is happening--a trend sometimes overlooked by physicians. Goldsmith says, "The key thing physicians have missed is that the patient is in charge of the process.... The Internet has enabled patients to aggregate their collective experience across disease entities." But there is too much information. "It is almost universally acknowledged by patients and physicians that there is a terrible quality problem. Getting from information to knowledge is a huge commercial opportunity for somebody." He doesn't think that people have put enough emphasis on the collective learning part of this new technology. PMID- 10788110 TI - The information revolution: opportunities and pitfalls for patients and providers. AB - The Information Revolution is changing everything in our world. Although this era is developing and evolving, some have described networks, globalization, diffusion of power away from professionals, and information overload as direct results of the Information Revolution. These megatrends are affecting every person and every organization in the world, and this article examines them to see how they might affect medicine and the delivery of health care in the millennium. What is truly significant about the changes brought about by the creation of cyberspace is that through computers, digitized information can be inexpensively and quickly transmitted anywhere in the world. Many of the traditional ethical and professional standards of medicine are feeling the strain of being applied to the strange, unfamiliar world-cyberconsultations and other virtual medical applications are explored. Physicians who understand this revolution will have a far better chance of controlling their destinies--and more importantly, aiding their patients in the quest for good health. PMID- 10788111 TI - Drowning in information. PMID- 10788112 TI - The urgent call for knowledge management in medicine. PMID- 10788113 TI - Paving the way for electronic medical records. AB - This article explores the reasons why electronic medical records have not become widely deployed in the health care industry. Y2K moved to center stage and fears of computer meltdowns became the single greatest obstacle to overcome before considering new technology possibilities. Almost every other information technology initiative in health care was delayed or suspended, while the issues of compatibility with four-digit dates were examined. Finding systems and devices that weren't Y2K compliant and replacing or working around their deficiencies left precious few resources available for other tasks. Another issue is standardization. The potential and promise of electronic records can only be realized if a standard way to describe clinical information can be agreed upon and implemented in practice situations. Clients have been reluctant to purchase electronic records while vendors are offering nonstandard solutions. Obstacles and benefits to implementing EMR systems are outlined. Despite some of the barriers to implementing EMRs, the future is bright for their widespread deployment. PMID- 10788114 TI - Health care information technology a primitive affair. PMID- 10788115 TI - The age of the Internet: working through the grief cycle (again). PMID- 10788116 TI - The clinical e-mail explosion. AB - The computer is transforming patient-physician communication. Physicians are already using electronic mail (e-mail) in physician-to-physician consultation, medical journal dissemination, and hospital-physician communication. This article addresses the value of e-mail for patient-physician communication. It explores the nature of this electronic medium, outlines published guidelines, delineates expected benefits and potential complications, and proposes how to incorporate e mail into health information systems. This article is designed to help guide clinicians and health care delivery organizations in the use of e-mail with patients to enhance rather than complicate the provider-patient relationship. Unique advantages of e-mail in the clinical setting include: the ability to offer routine transactions and patient education; increased efficiency; the self documenting nature of this medium; cost-effectiveness; and serving as a clinical extender. PMID- 10788117 TI - Building a framework to transform health care. AB - Advances in information technology are helping clinicians to realize the promise of evidence-based medicine, which includes benchmarking, outcomes monitoring, predictive modeling, and clinical pathways. By integrating individual clinical expertise and the best available research, physicians can apply the disciplines and techniques of clinical research to their practice of medicine, one patient at a time. Evidence-based medicine also allows organizations to move forward with continuous clinical quality improvement programs. Standards, open systems, data warehouses, and evidence-based medicine help a health care delivery system obtain the technical infrastructure, decision-making processes, analytical skills, clinical databases, predictive models, and clinical pathways. With this information technology (1) physicians can practice evidence-based medicine and (2) the delivery system can profile clinicians' practice habits for managed care contracting and continuous clinical quality improvement. PMID- 10788118 TI - Emerging from the cybermaze. PMID- 10788119 TI - Are you ready for generation X? AB - Generational analysis considers the differences in world view and attitude between various generations of Americans and uses this information to develop insight and business strategies. The much discussed generation gap between the Baby Boomers and their elders during the 1960s and 1970s is equaled or exceeded by the differences between Generations X and Y and the Boomers. Generation X physicians will be coming into practice over the next two decades. Successful medical leaders must understand Gen-Xers to create clinical environments that meet their unique needs and concerns. This article describes some of the philosophical and attitudinal differences between the generations and what these differences may mean to the future of medicine and health care organizations. PMID- 10788120 TI - Working with a net. AB - Networking is a critical skill-set that physicians, as they evolve into management and executive positions, need to develop and utilize. A network is a web of personal and professional connections built up over a lifetime based on mutual interests and nurtured by ongoing regular contacts. Good networkers, most especially those for whom bonhomie is not second nature, pursue the project systematically. They set up a schedule for keeping connections alive, they compile and refresh the network database, they attend forums where old acquaintances can be renewed and new ones formed, they follow a set of best practices, and they arm themselves with the essential instruments of the networking trade (primarily a pencil, a business card, and an open attitude). Successful networking requires the ability to ask for favors--but also to extend them willingly. Networking is an exponential process of harnessing connections to connections--one's own network to those of others, to gain knowledge and elicit opportunities that would otherwise be unavailable. PMID- 10788121 TI - Believing is seeing. AB - Useful, well-demonstrated, well-vetted ideas in clinical practice, disease management, health care management, ideas that would save lives, save money, and make life better for the patient, are sometimes simply ignored, dismissed as radical, as completely unfounded, dangerous, and without merit. Why are new ideas so slow to spread in medicine and health care? Because believing is seeing. We do not look for something we don't believe in. In fact, we do not even see a thing if we don't believe in it. We have dedicated ourselves so powerfully to medicine, to health care as we know it, that we often do not even see any alternatives. A combination of factors makes it likely that, in the coming decade or two, we will change almost everything that is fundamental about health care and medicine. In a time of such rapid change, we desperately need to root out and question our deep assumptions and beliefs, to get off the tracks laid down by training and experience and ask questions we have never asked before. PMID- 10788122 TI - Managing the deliberately mute. AB - The contrast in communication styles and values between Gen-Xers, now mostly in their mid to late twenties, and forty- and fifty-somethings is obvious. Gen-Xers are focused on the assignment and the deadline; their goal is to do good work in a timely manner. But they are highly skeptical that enthusiasm has any influence on the outcome. When we question them about their taciturn manner, they all give us the same two reasons: They really don't care one way or the other and they're convinced that what they say doesn't matter anyhow. This may frustrate a manager charged with getting the buy-in or enthusiastic participation from the troops, but it's a fact. There are, however, ways to get Xers to talk--provided you really want their ideas and opinions and you acknowledge that you hear what they say. Here are the best techniques from those who successfully manage large numbers of the young, including young physicians: (1) Focus on what matters; (2) don't ask if you're not going to act on the feedback; (3) personalize your request for information; and (4) always do a worst case scenario when you need the buy-in. PMID- 10788123 TI - Mentoring: women learning from others. AB - Informal mentoring allows a relationship to evolve naturally over time much like a friendship, without an official obligation or commitment to coach someone. However, some women prefer a more formal, organizationally driven approach to mentoring. In either case, both represent an opportunity to learn and grow. In Learning from Other Women, Carolyn Duff interviewed many women to understand their expectations, preferences, and experiences. Mentoring, Duff says, "begins with affinity between two people, but the focus remains around work. It's a magical thing that happens when one person sees something in another person and wants to help that person grow." This column explores some of the barriers, as well as how to ask for help and benefit from a mentor. Whether you like the word "mentoring" or not, whether your organization has a formal program or not, always be thinking about how you can learn from others. Keep your eyes and mind open, look around to see who is doing what you want to do. PMID- 10788124 TI - Getting started as a physician executive. AB - In clinical practice, technical skills often outweigh interpersonal and leadership skills as success factors--you can be a great doctor and a so-so person. But the reverse seems to be true in the physician executive role; it is precisely the intangible leadership skills that contribute to and determine potential success. And they can be tough to master, especially when you focus on them for the first time, partway through an already-successful career. Practicing leadership is like practicing medicine. It's not just a matter of learning some new things--if it were only that, physicians are known to be excellent learners. Nor is it just a matter of determination or application--this is not a battle that sweat and effort alone can win. Most physicians will want to "try on" the executive role before making strategic moves in that direction. But be clear about what you want to get out of any project or activity before you jump in. If you're seeking a management degree, the best approach is to tie together developing your technical and interpersonal skills, as well as the formal credentials. PMID- 10788125 TI - The age of accountability. AB - The advent of accountability and evidence-based medicine set a new standard, suggesting that new medical technologies should require validation of their safety and effectiveness through controlled studies prior to their broad dissemination. Certainly, the concept of evidence-based medicine appears to be rational, objective, and consistent. However, the process can become extremely messy in its application to coverage decisions. After all, the coverage decision is really one of the most basic elements of health care. The following discussion represents a week in the life of a mythical medical director who is attempting to implement his or her company's evidence-based coverage policies. PMID- 10788126 TI - The digital transformation of health care. AB - The arrival of the Internet offers the opportunity to fundamentally reinvent medicine and health care delivery. The "e-health" era is nothing less than the digital transformation of the practice of medicine, as well as the business side of the health industry. Health care is only now arriving in the "Information Economy." The Internet is the next frontier of health care. Health care consumers are flooding into cyberspace, and an Internet-based industry of health information providers is springing up to serve them. Internet technology may rank with antibiotics, genetics, and computers as among the most important changes for medical care delivery. Utilizing e-health strategies will expand exponentially in the next five years, as America's health care executives shift to applying IS/IT (information systems/information technology) to the fundamental business and clinical processes of the health care enterprise. Internet-savvy physician executives will provide a bridge between medicine and management in the adoption of e-health technology. PMID- 10788127 TI - Chemical and biological terrorism: planning for the worst. AB - Preparing for a chemical or biological terrorist event has recently become an intense area of focus for emergency health planners in every state and city in America. Many of these individuals believe that a chemical or biological terrorist event is likely in the near future and they raise concerns about the readiness of our nation's health system to rapidly identify and manage a disaster of this magnitude. In both types of events, physician leadership is essential to reduce death and disability and restore public order. Several policy issues need to be addressed in this complex issue. Physician executives play a key role in preparing the health system to respond to this type of disaster. PMID- 10788128 TI - A scourge of drugs strikes a pious place. Addiction grows among Orthodox Jewish youth. PMID- 10788129 TI - The perils of pills. The psychiatric medication of children is dangerously haphazard. PMID- 10788130 TI - Vision loss in seniors: the future looks brighter. PMID- 10788131 TI - While diet docs debate fats and carbs, weight loss still comes down to calories. PMID- 10788132 TI - Employers boxed in as benefit costs rise. PMID- 10788133 TI - Proving that ethics are efficient. PMID- 10788134 TI - Washington wakes up to medical mistakes. PMID- 10788135 TI - The high cost of couch potatoes. PMID- 10788136 TI - A new boost for benefits management. PMID- 10788137 TI - What we've learned about carving out health care. PMID- 10788138 TI - Can old Blues learn new tricks? PMID- 10788139 TI - The lowdown on those new COBRA regs. PMID- 10788140 TI - Not just another rate increase. PMID- 10788141 TI - Data watch. An ounce of prevention. PMID- 10788143 TI - Rising health insurance costs. PMID- 10788144 TI - 85% of lenders won't lend to healthcare companies. PMID- 10788142 TI - Market dynamics. Part III: Will physicians survive managed care? PMID- 10788145 TI - Class action suits against HMOs multiply. PMID- 10788146 TI - AMA survey shows increased physician use of the Internet. PMID- 10788147 TI - Medical errors kill 95,000 a year. PMID- 10788148 TI - The nature of nurturing. A new study finds that how parents treat a child can shape which of his genes turn on. PMID- 10788149 TI - A very tiny ancestor. Scientists fill in a piece of the evolutionary puzzle. PMID- 10788150 TI - Drugstore dangers. Medication mix-ups are increasing as the number of prescriptions grows each year. Here are the hazards, and the steps you can take to protect yourself. PMID- 10788151 TI - Mediation before malpractice suits? For patients, litigation is expensive and hard to win. The case for trying talking instead of suing. PMID- 10788152 TI - Coping with the Health Insurance Portability and Accountability Act of 1996. PMID- 10788153 TI - HIPAA creates an opportunity to rethink IT strategy. PMID- 10788154 TI - HIPAA checklist: getting organized, informed, and proactive. PMID- 10788155 TI - White House to propose more money for survey and certification; industry claims funding is misdirected. PMID- 10788156 TI - MedPAC to recommend simplification of data collection, better coordination across settings. PMID- 10788157 TI - Court ruling could help providers hit with False Claims Act investigations. PMID- 10788158 TI - How to reduce nursing costs without adversely affecting quality. PMID- 10788159 TI - Mariner becomes third national SNF chain to file for bankruptcy protection since Medicare PPS implementation. PMID- 10788160 TI - Physician collective bargaining and federal antitrust law. PMID- 10788161 TI - Qui tam on the lam: Fifth Circuit fires the first salvo--will the Supreme Court be next? PMID- 10788162 TI - Any willing provider laws for pharmacies. PMID- 10788163 TI - Analyzing the multiple indicators of customer loyalty. PMID- 10788164 TI - Addressing the new healthcare superpower. PMID- 10788165 TI - The gene hunters. Unlocking the secrets of DNA to cure disease, slow aging. PMID- 10788166 TI - Betsy Dresser. Saving wildlife with frozen zoos--and cloning. PMID- 10788167 TI - Yuan Chang and Patrick Moore. Sharing an office and a marriage, they hunt infectious agents. PMID- 10788168 TI - Nan Unklesbay. Making hamburgers safe for humankind. PMID- 10788169 TI - Ilya Raskin. Probing the roots of plant-based drugs. PMID- 10788170 TI - Philip Kennedy. Melding man and machine to free the paralyzed. PMID- 10788171 TI - Rodolfo Llinas. A grand unification theory of the brain. PMID- 10788172 TI - Marc Tessier-Lavigne. Training nerve cells to mend a damaged spine. PMID- 10788173 TI - Add AIDS to Clinton's legacy. PMID- 10788174 TI - A tailor-made vaccine. Scientists try a new strategy to combat cancer. PMID- 10788175 TI - No sex for the serious. The smartest go slowest. PMID- 10788176 TI - Healing the heart. Within just a few years, new weapons against heart disease will be working to stamp out the nation's leading killer. PMID- 10788177 TI - The 'man's illness' can be deadlier to women who don't spot cardiac trouble. PMID- 10788178 TI - These winners HAVE it all--year 2000 Hospital Awards for Volunteer Excellence. PMID- 10788179 TI - Fighting fire with cease fire. Thoughts on guns and health care. PMID- 10788180 TI - Living with pain. Pass on this advice to those you know in chronic pain. PMID- 10788181 TI - The alternative to tube-feeding patients with advanced dementia. PMID- 10788182 TI - Takin' it to the Hill. Volunteers make their voice heard in Washington. PMID- 10788183 TI - A grab bag of good ideas. Making the most of the occasional volunteer. PMID- 10788184 TI - Dazzling images: innovative recruitment strategies. PMID- 10788185 TI - Predictors of long-term weight reduction in obese patients after initial very-low calorie diet. AB - This prospective interventional study assessed possible predictors of long-term weight loss and compared them with factors previously identified as predicting short-term variations in weight reduction after initial treatment with a very-low calorie diet (VLCD). Eighty-two overweight patients with a body mass index of at least 27 kg/m2 were recruited from primary health-care settings into a structured weight-reduction and maintenance program. All patients used the VLCD for 8 weeks and were followed up 13.2 months later. Mean body weight decreased an average of 13.3 kg during the 8-week treatment and was still 8.6 kg below pretreatment levels after 13.2 months. Triglyceride levels were also significantly reduced. Sex, baseline weight, baseline body mass index, and age predicted 37% of the variation in short-term weight loss but had no long-term predictive value. PMID- 10788186 TI - Is rehab in trouble? PMID- 10788187 TI - Update on wound care. PMID- 10788188 TI - Understanding MDS. PMID- 10788189 TI - The core essentials. PMID- 10788190 TI - Important lessons to learn. PMID- 10788191 TI - Long-term rehab. Improving quality of life. PMID- 10788192 TI - Making the right choice. PMID- 10788193 TI - Seating and positioning in the newly injured. PMID- 10788194 TI - Breaking barriers. PMID- 10788195 TI - Boosting performance. PMID- 10788196 TI - Tailoring: what's in a name? PMID- 10788197 TI - Promising community-level indicators for evaluating cardiovascular health promotion programs. AB - Rigorous evaluation of community-based programs can be costly, particularly when a representative sample of all members of the community are surveyed in order to assess the impact of a program on individual health behavior. Community-level indicators (CLIs), which are based on observations of aspects of the community other than those associated with individuals, may serve to supplement individual level measures in the evaluation of community-based programs or in some cases provide a lower-cost alternative to individual-level measures. Because they are often based on observations of the community environment, CLIs also provide a way of measuring environmental changes--often an intermediate goal of community-based programs. The Centers for Disease Control and Prevention convened a panel of experts knowledgeable about community-based program evaluation and cardiovascular disease (CVD) prevention to develop a list of CLIs, and rate their feasibility, reliability and validity. The indicators developed by the panel covered tobacco use, physical activity, diet and a fourth group that were considered 'cross cutting' because they related to all three behaviors. The indicators were subdivided into policy and regulation, information, environmental change, and behavioral outcome. For example, policy and regulation indicators included laws and ordinances on tobacco use, policies on physical education, and guidelines for menu and food preparation. These indicators provide a good starting point for communities interested in tracking CVD-related outcomes at the community level. PMID- 10788198 TI - Autonomy, health and ageing: transnational perspectives. AB - A comparative study was undertaken in Italy and the UK to explore elderly people's perceptions of old age and ageing, and to establish a ranking of factors which were seen to contribute to the maintenance or loss of autonomy. The results were collated with the expressed views of practitioners and others working with elderly people in a range of settings in eight different European Union Member States. These data informed the compilation of an educational programme, presented as a handbook for use by and with elderly people, the focus of which was life-skills development as a prerequisite for health education. Life-skills were defined in terms of the development of a positive self-image, a social 'ease' and a feeling of 'belongingness' in the context of old age. Assumptions underlying the framing of the educational programme were a transnationally accepted relationship between autonomy, empowerment, self-image and health, and the centrality of life-skills development as catalytic in this process. The paper, however, flags substantial conceptual and methodological issues which arose in moving towards transnationally, shared understandings within the project team at each of the three stages of the project, and offers some evaluative observations on the strengths, concerns and achievements offered by transnational research and collaborative activity. PMID- 10788199 TI - From psycho-social theory to sustainable classroom practice: developing a research-based teacher-delivered sex education programme. AB - This paper describes the development of a theoretically based sex education programme currently undergoing a randomized controlled trial in the UK. It considers some of the practical difficulties involved in translating research based conclusions into acceptable, replicable and potentially effective classroom lessons. The discussion acknowledges that the implications of social psychological research and the requirements of rigorous evaluation may conflict with accepted principles inherent in current sex education practice. It also emphasizes that theoretical ideas must be carefully embedded in lessons which are informed by an awareness of classroom culture, and the needs and skills of teachers. For example, the use of same-sex student groups to reflect on the gendered construction of sexuality may be problematic. Materials must be tailored to recipients' circumstances, which may require substituting for limited experience with the use of detailed scripts and scenarios. Furthermore, role-play techniques for sexual negotiation that work elsewhere may not be effective in the UK. The use of trigger video sessions and other techniques are recommended. Finally, the problems involved in promoting condom-related skills are discussed. The paper concludes that, if an intervention is to be sustainable beyond the research stage, it must be designed to overcome such problems while remaining theoretically informed. PMID- 10788200 TI - An experimental investigation of the influence of health information on children's taste preferences. AB - Promotion of healthy diets often involves provision of information about which food types are most favourable for health. This is based on the assumption that the rational consumer will, other things being equal, choose the food that they know is healthier. However, health information may not always have a positive effect, since there is evidence that some people, particularly children, believe that healthiness and tastiness are mutually exclusive characteristics. To the extent that taste governs preferences and consumption, the characterization of a food as healthy could reduce its anticipated pleasantness. The present study tested the idea that a 'healthy' label would reduce liking for a novel drink. The results showed that the children rated a 'healthy labelled' drink as less pleasant and said they would be less likely to ask their parents to buy it than the same drink presented with control information. These results suggest that care may need to be exercised in promoting foods to children through an emphasis on health, unless the implications of healthiness can be rendered more positive. PMID- 10788201 TI - Outcome evaluation of a multi-component violence-prevention program for middle schools: the Students for Peace project. AB - This study evaluated the effect of Students for Peace, a multi-component violence prevention intervention, on reducing aggressive behaviors among students of eight middle schools randomly assigned into intervention or control conditions. The intervention, based on Social Cognitive Theory, included the formation of a School Health Promotion Council, training of peer mediators and peer helpers, training of teachers in conflict resolution, a violence-prevention curriculum, and newsletters for parents. All students were evaluated in the spring of 1994, 1995 and 1996 (approximately 9000 students per evaluation). Sixth graders in 1994 were followed through seventh grade in 1995 or eighth grade in 1996 or both (n = 2246). Cohort and cross-sectional evaluations indicated little to no intervention effect in reducing aggressive behaviors, fights at school, injuries due to fighting, missing classes because of feeling unsafe at school or being threatened to be hurt. For all variables, the strongest predictors of violence in eighth grade were violence in sixth grade and low academic performance. Although ideal and frequently recommended, the holistic approach to prevention in schools in which teachers, administrators and staff model peaceful conflict resolution is difficult to implement, and, in this case, proved ineffective. The Students for Peace experience suggests that interventions begin prior to middle school, explore social environmental intervention strategies, and involve parents and community members. PMID- 10788202 TI - Knowledge and attitudes toward AIDS among female college students in Nagasaki, Japan. AB - This study assesses knowledge and attitudes concerning HIV infection and individuals with AIDS among 383 female students attending colleges in Nagasaki, Japan. A structured questionnaire containing questions concerning knowledge about AIDS, sources of information, beliefs and attitudes toward people with HIV/AIDS was administered during sessions set up for that purpose. The mean age of participants was 18.8 +/- 0.8 years (+/- SD). The main source of information for AIDS awareness as reported by the students was the mass media. Good knowledge about AIDS was positively associated with ease of acceptance of living in the same house with a person diagnosed with AIDS [odds ratio (OR): 1.90; 95% confidence interval (CI): 1.07-3.38]. However, residing at home (OR: 0.64; 95% CI: 0.42-0.98) and involvement in nurse education programmes (OR: 0.59; 95% CI: 0.37-0.95) showed a negative association. Students demonstrated a high level of knowledge concerning AIDS and HIV, but had considerable misconceptions and prejudices about people having HIV/AIDS. Our results suggest that a more appropriate education programme in colleges in Japan may be necessary to reduce the discrepancy between general knowledge and desirable attitude regarding HIV/AIDS. PMID- 10788203 TI - The Western Australian School Health Project: comparing the effects of intervention intensity on organizational support for school health promotion. AB - The aim of this study was to evaluate changes in school health promotion practice related to two levels of intervention in the Western Australian School Health (WASH) Project: (1) a low-intensity intervention involving a single mail-out of WASH Project resources, and (2) a high-intensity intervention involving training, planning time and expert support. The schools involved in the study were divided into three groups. Treatment group 1 received the high-intensity intervention, treatment group 2 received the low-intensity intervention and a comparison group received no intervention. Two scales were developed to assess change, i.e. a school organizational scale (Chronbach's alpha = 0.76) and a health promotion activity scale (Chronbach's alpha = 0.79). The results indicate that a high intensity intervention, such as the WASH Project, which provides training to a critical mass of school community members from each school, ongoing access to an expert in the field, as well as dedicated planning time, is able to increase the comprehensiveness and quality of health strategic planning by schools. Furthermore, the results suggest that a low-intensity mail-out intervention is no more successful in initiating change that providing no intervention at all. PMID- 10788204 TI - Evaluating prevention programs with the Results Mapping evaluation tool: a case study of a youth substance abuse prevention program. AB - The harmful effects of alcohol and other drug abuse are widespread. Our health care, social service, education and legal systems are strained under the impact of substance abuse, not to mention the economic costs associated with substance abuse. Consequently, effective strategies which prevent substance abuse must be identified and replicated. Yet, user-friendly and cost-effective evaluation tools for community-based substance abuse prevention programs are rare. A recently developed tool that has promise to overcome some of the barriers which exist when evaluating prevention programs is 'Results Mapping'. Results Mapping documents and aims to quantify the contributions of a program to future outcomes of its target population with the intention of making data meaningful at the program and funding agency level. This case study was conducted to assess the feasibility of implementing the Results Mapping evaluation tool for community-based prevention programs. The study assessed qualitatively how well Results Mapping worked for one community-based substance abuse prevention program, as well as how much time and funding it took to implement this new tool. Results suggested that Results Mapping may be a valuable documentation, planning and learning tool, but funding agencies should be cautious about using Results Mapping scores to determine funding allocations. PMID- 10788205 TI - Understanding peer education: insights from a process evaluation. AB - In the UK, peer education has become an increasingly popular way of carrying out health promotion work with young people but evaluations of its effectiveness remain largely unpublished. In particular, illuminative evaluations using qualitative methods are rarely reported both in the UK and other countries. This paper presents insights from the process evaluation of a peer education project in Fife, Scotland which was funded to explore new ways of working with young people in the areas of sexual health, HIV/AIDS and drugs. The interactive approach of the evaluation and its responsiveness to the development of the project are outlined. Factors influencing the peer education process, such as recruitment, setting, organizational context and personal development of participants, are described. Aspects of the formal and informal work carried out by the peer educators are discussed. It is hoped that the paper may provide a starting point for developing more reflective understandings about the processes involved when peers educate peers. PMID- 10788206 TI - Internalized homophobia and health issues affecting lesbians and gay men. AB - This paper investigates the concept of internalized homophobia in both theory and research relating to lesbian and gay health. It offers a contemporary and critical review of research in this area, and discusses a range of recent findings relating to a range of health issues including HIV and AIDS. Whilst the concept has a resonance for gay men and lesbians, and is widely used in 'lesbian and gay-affirmative' interventions, the paper demonstrates that research findings have been equivocal and the term is often used without full consideration of its sociopolitical consequences. The paper concludes that the concept does have a valuable role to play in health promotion work with lesbians and gay men but invites further discussion and examination of the construct. PMID- 10788207 TI - New physicians frustrated by coding, managed care, journal deluge. PMID- 10788208 TI - What lies ahead for family physicians? PMID- 10788209 TI - Practicing in the new millennium: do you have what it takes? PMID- 10788210 TI - The emergence of consumer-driven health care. PMID- 10788211 TI - The family physician's role in reducing medical errors. PMID- 10788212 TI - Can patient demand for pharmaceuticals be moderated? AB - OBJECTIVE: Ask health plan members whether they would consider switching to a less costly medication or alternative treatment if their physician asked. SAMPLE & SETTING: Random sample of 265 members of Kaiser Permanente in Northern California. DESIGN: A telephone survey asked participants to respond to three vignettes tailored to either men and women age 18 to 39, women age 40-80, or men age 40-80. MAIN RESULTS: If asked to do so by their doctors, most young adults (77%) said they would be open to switching to a less expensive allergy medicine, most women over age 40 (60%) said they would try a less expensive blood pressure medication, and most men over age 40 (68%) said they would meet with a behavioral specialist and try other steps before taking ViagraAE. In open feedback, participants said they would be most receptive if the recommendation came from a trusted physician, if given the freedom to choose otherwise, and the alternative was equally effective and convenient. About one-third of participants expressed reservations or said they would definitely refuse substitution. These refusers were significantly less satisfied with the health plan, less satisfied with their primary care doctor, less trusting of Kaiser physicians in general, and in poorer health (as indicated in one or more vignettes). CONCLUSIONS: Moderating patients' demand for pharmaceuticals can begin by physicians skillfully asking patients to consider a more cost-effective alternative to expensive medications when one is available. PMID- 10788213 TI - The devolution of critical care in the U.S. PMID- 10788214 TI - The cost and quality of hospitalists. AB - The Hospitalist concept is becoming the standard of practice as managed care penetration grows. It is increasingly difficult to do a good job in the outpatient arena and this makes the old "double threat" model in which the physician speeds through inpatient rounds in the morning before clinic, during lunch, and again at night after his office closes, obsolete. There is simply too much at stake in the inpatient arena to have large periods of time in which physician coverage is minimal. Controversy exists as to whether or not the services should be "outsourced" to a Hospitalist management company or developed internally, using physicians who historically have had a significant presence in the hospital. If the services are outsourced, then it is essential that they be built around a strong local physician leader who remains active in patient care. The French author, Anais Nin wrote: "There are very few human beings who receive the truth, complete and staggering, by instant illumination. Most of them acquire it fragment by fragment, on a small scale, by successive developments, cellularly, like a laborious mosaic." Inpatient care in the United States is adequately described as a laborious mosaic in which providers of many different services surround patients with many different problems. Hospitalist programs can help to bring order to the mosaic by consistently combining the correct physician talent with the system's sickest and most expensive patients in order to achieve the best possible outcomes. PMID- 10788215 TI - Academic medicine and managed care: a unique collaboration. PMID- 10788216 TI - Consumerism and health care quality. PMID- 10788217 TI - Breach of fiduciary duty. PMID- 10788218 TI - Developing an e-business strategy. PMID- 10788219 TI - Standards and procedures for determining whether a defendant is competent to make the ultimate choice-death; Ohio's new precedent for death row "volunteers". PMID- 10788220 TI - Race for perfection: children's rights and enhancement drugs. PMID- 10788221 TI - Pitfalls in diagnosis of occupational lung disease for purposes of compensation- one physician's perspective. AB - Occupational caused lung disease (OLD) is almost always compensable, either by application to workers' compensation agencies or by a civil lawsuit. For this reason the diagnosis usually comes under close scrutiny. Several pitfalls can occur when a physician diagnoses a patient as having lung disease of occupational origin, especially when compensation is at issue. These pitfalls can trap both the attorney advocate and his client, and lead to a result opposite of that intended (e.g., the claim one supports can be denied). For purposes of discussion I have categorized the pitfalls as follows: A. Making an unsupported medical diagnosis (looking for a "quick fix"). B. Echoing an unsupported diagnosis made by someone else. C. Inadequate clarification of 'impairment' and 'disability.' D. Ignoring or minimizing relevant medical history. E. Arguing against yourself. F. Ignoring the possibility of a rare or unusual diagnosis. G. Not obtaining or reviewing independent chest x-rays and reports. H. Attributing causation with certainty when it is unwarranted by the facts. I. Relying on a claimant's own smoking history. J. Misinterpreting pulmonary function and arterial blood gas tests. K. Missing the real cause of a patient's complaint. L. Diagnosing occupational lung disease without attempting to remove the patient from the cause. M. Confusion over basic terminology and pathophysiology in OLD. N. Using sloppy or incorrect language, including misspelling. O. Not saying "I don't know," when you don't know. Pitfalls in diagnosis generally arise from either physician bias or inadequate evaluation. Although most pitfalls seem to be made by physicians on the plaintiff's side, they are also made by physicians on the defendant's side, as when bias interferes with recognizing a condition that is occupational in origin. Ideally, the fact that diagnosis of OLD involves the legal profession should not affect a physician's objectivity or clinical approach. Physicians have an obligation to help assure that deserving patients receive compensation, and that claimants without a compensable occupational illness are not unjustly rewarded. However, the attorney's need to prove a diagnosis "with medical certainty," and the defendant's need to refute that diagnosis with equal certainty, often skew what would otherwise be a straightforward diagnostic process. Resulting pitfalls in diagnosis can, in the end, trap the physician advocate and the side he is trying to help. PMID- 10788222 TI - International response to Dolly: will scientific freedom get sheared? PMID- 10788223 TI - The working uninsured: a lucrative market for innovative providers. PMID- 10788224 TI - Marketing best practices in alternative medicine. AB - Healthcare organizations throughout the country are endeavoring to select the best strategy to pursue in the area of alternative medicine. In an effort to define and clarify choices, the Alliance for Healthcare Strategy and Marketing has just released the second in its series of Marketing Best Practices research reports. This report documents the marketing "best practices" of 15 organizations or alternative medicine consultants throughout the country (see figure 1). The study revealed some distinct trends and practices that can be a predictor of success for other programs. PMID- 10788225 TI - State action antitrust immunity for public hospitals: it depends on what you mean by "foreseeable". AB - The Supreme Court's refusal to hear an appeal from the Fifth Circuit's ruling on the application of state action antitrust immunity doctrine leaves a split in the circuits as to the proper test to apply to determine antitrust immunity for public hospitals. The circuits have either adopted a "foreseeable conduct from state policy" approach or a "policy from foreseeable conduct" approach. This Article analyzes the approaches and demonstrates that the "foreseeable conduct from state policy" approach is the better of the two methods. PMID- 10788226 TI - Hospital mergers: the shift from federal antitrust enforcement to state regulation. AB - The authors examine and analyze the burgeoning merger activity in the hospital arena, as well as the nonfederal attempts made to regulate that activity. They conclude that the present, ad hoc, system of state regulation is sorely wanting and that it would be preferable if stronger antitrust enforcement and judical decisions prevented competition reducing mergers. If a merger results in a true monopoly (and nonetheless passes antitrust scrutiny), its regulation should be the responsibility of the pertinent state public utility board which, unlike the courts and state attorneys general, has sufficient expertise to adequately regulate the merged entities. Otherwise, the faults of the present system, which is easily manipulated by hospitals seeking political and legal cover for their activities, are likely to be perpetuated. PMID- 10788227 TI - FDA regulation of medical software. AB - Currently the FDA is reviewing its policy on the regulation of computer software in the medical arena. Healthcare practitioners hope that the FDA will take the least possible regulatory action required by law to fulfill its public health responsibility and will take into account the unique status of the ever-changing software industry. Innovations in telemedicine and medical informatics have yielded significant improvements in the quality and cost of healthcare, and the imposition of cumbersome regulatory requirements on the medical software industry would slow the development and introduction of medical software products. PMID- 10788228 TI - Physician collective bargaining: all eyes are on Texas. AB - In June of 1999, Texas became the first state to pass a law allowing physicians to collectively bargain with managed care plans for fee-related activities. Whether this law will give physicians renewed control over patient care remains to be seen. Notwithstanding the intention of its drafters, it is likely that the law and its proposed regulations may only provide physicians with costly and time consuming procedures that present multiple risks instead of renewed power. PMID- 10788229 TI - Sample patient transfer policy. PMID- 10788230 TI - EMTALA: reaching beyond the emergency room to expand hospital liability. AB - The Office of the Inspector General and private plaintiffs are vigorously pursuing EMTALA violations. These efforts are particularly troubling to hospitals, who face difficult statutory interpretation and application questions, especially in light of managed care reimbursement requirements. Two recent cases, one of them from the United States Supreme Court, expand hospital liability under EMTALA. This Article reviews current EMTALA standards and regulations, and analyzes how the recent decisions, in the absence of congressional action, are likely to impose substantial burdens upon hospitals. PMID- 10788231 TI - Applying antitrust law to nonprofit healthcare entities: arguments for a greater attention to detail. AB - This Article begins with an antitrust primer, then analyzes the appropriate application of antitrust principles to nonprofit healthcare providers. In light of the inherent charitable character of nonprofit healthcare providers, the author contends that the government and the courts should accord some deference to nonprofit hospitals when they are seeking approval of mergers. To date, this has not generally been the case, although a few recent court decisions have rested their approval of mergers, in part, upon the nonprofit character of the merging entities. The author, in particular, believes the paradigmatic local nonprofit hospital with a community board is less likely than a for-profit hospital to abuse any market power that it may obtain through a merger; consequently, any such merger should not be analyzed solely under the traditional presumptions of antitrust jurisprudence. Rather, the premerger analysis should involve meaningful consideration of the hospital's charitable character. PMID- 10788232 TI - A colleague may be losing it--but asks you not to tell. Roundtable discussion. PMID- 10788233 TI - Thar's gold (maybe) in those ancillary services. PMID- 10788234 TI - The patient had a headache, and I didn't feel so well myself. PMID- 10788235 TI - How alternative providers get their credentials. PMID- 10788236 TI - William G. Louis, MD. From high school dropout to teacher of youth. PMID- 10788237 TI - Are you sure your patient can read? PMID- 10788238 TI - The people who shaped our world. PMID- 10788239 TI - Electronic records--I can't live without 'em. PMID- 10788240 TI - Freshen your practice--make house calls. PMID- 10788241 TI - Capitation on the rise. PMID- 10788242 TI - I tell patients, "Pay the piper". PMID- 10788243 TI - Fraud and abuse. Government agencies have a new weapon in their arsenal. PMID- 10788244 TI - Physician recruitment. Specialists find the warmest welcome in years. PMID- 10788245 TI - Say goodbye to your med school debt. PMID- 10788246 TI - Anatomy of a killer's hand. PMID- 10788248 TI - The e-train is coming at you. PMID- 10788247 TI - What's in a name? For DOs, plenty. PMID- 10788249 TI - Can doctors ever make a profit owning a hospital? PMID- 10788250 TI - You can't be the right doctor for your family. PMID- 10788251 TI - Tobacco wins in court. The FDA can no longer monitor sales to kids. PMID- 10788252 TI - When a pest turns menace. Be on the lookout for more outbreaks of deadly mosquito borne disease. PMID- 10788253 TI - The wisdom of the gut. Those butterflies in your stomach are not just in your mind. PMID- 10788254 TI - Teen angels, can you hear this message? Young drivers plus passengers equals disaster. PMID- 10788255 TI - Telltale test for the heart. A key protein tracks inflamed arteries. PMID- 10788256 TI - Prostate cancer: kinder cuts mean sharper dilemmas. PMID- 10788257 TI - Re-engineering the role and education of clinical laboratory scientists. PMID- 10788258 TI - Curricula alternatives in diagnostic laboratory techniques. PMID- 10788259 TI - Reverse transcriptase PCR quantitation of hepatitis C virus. AB - OBJECTIVE: To present a brief review of the diagnostic benefits of quantitating viral load for hepatitis C and how the reverse transcriptase polymerase chain reaction is being used as an aid to better diagnose and manage the disease. DATA SOURCE: Research articles about hepatitis C and the reverse transcriptase polymerase chain reaction, as well as data gathered by the authors. STUDY SELECTION: Performed by the authors. DATA EXTRACTION: Performed by the authors. DATA SYNTHESIS: Hepatitis C viral infection is a worldwide health problem, affecting about 100 million people worldwide. Numerous serological tests exist to detect antibodies to hepatitis C antigens, but some affected people fail to generate an immune response. Reactivity in the reverse transcriptase polymerase chain reaction is definitive proof of hepatitis C infection. The titer of RNA indicates patient response to antiviral therapy. Measuring the presence and quantity of RNA by the reverse transcriptase polymerase chain reaction has become an important aid for diagnosis and monitoring of hepatitis C infection. CONCLUSION: The reverse transcriptase polymerase chain reaction method is a highly sensitive and accurate aid in diagnosing or confirming diagnosis of hepatitis C viral infection. This method is widely used to assess likelihood of patient response to therapy, and to monitor efficacy during therapy. PMID- 10788260 TI - But can they do it? Clinical competency assessment. AB - There are four distinct steps in the development of an effective competency assessment process. Step 1 is the definition of competency criteria in consultation with prospective employers. Step 2 is the selection of assessment tools. The method or tool that will provide a comprehensive, reliable, valid and cost-effective measure of clinical competence does not currently exist; therefore the judicious selection of the right combination of evaluation methods based on predetermined competency criteria is crucial. Step 3 is implementation of the selected assessment tools, followed by Step 4, evaluation of the process and any changes in the work environment. Then the cycle is instituted again with redefinition of competency criteria if indicated. PMID- 10788261 TI - Phlebotomy skills expected of career entry CLS/CLT graduates: a Missouri hospital perspective. AB - OBJECTIVE: To determine how much, what type, and what proficiency of phlebotomy experience CLS/CLT students should have during the training program to be prepared to meet the needs of the majority of Missouri hospital employers. DESIGN: Survey to determine the role healthcare professionals, inside and outside the laboratory, play in today's blood collection patterns and phlebotomy management. SETTING/PARTICIPANTS: The Missouri Organization of Clinical Laboratory Science mailed 204 surveys to the Missouri Hospital Association member laboratories. MAIN OUTCOMES/CONCLUSIONS: This research examined the need for modifying phlebotomy skills of clinical laboratory science students. Data gathered from employers support the premise that entry-level competencies of CLS/CLT graduates will vary according to clinical facility size. CLS/CLT programs may use data from this study to plan phlebotomy practicums. It can be extrapolated that Missouri employers who are most likely to employ career entry graduates expect them to draw blood from 9.3 patients within one hour. Fifty three percent of 40 to 400 bed hospitals expect graduates to perform difficult draws in at least eight types of hospital units. Laboratories are the major managers of hospital wide phlebotomy services; thus, CLS/CLT curricula should include phlebotomy management methods. PMID- 10788262 TI - Imaging in the 21st century. AB - The drive for cost reduction and increased quality of therapy (patient outcome) will lead to increasingly less-invasive procedures. As a result, imaging will become essential in the operating theatre environment, simply because the site of the intervention is not visible to the clinician performing it. PMID- 10788263 TI - Conflict resolution and Y2K problems. How biomaterials will face up to the real issues of the future. AB - At the end of this century, the medical device industry has achieved some notable successes, yet achieves so little in terms of significant functional replacement and reconstruction of the body. This is partly because of the materials that are used. This article addresses the main points of concern and highlights the way in which medical devices of the future will be based on different principles and will deploy different tactics in the search for safe and reliable treatments of disease and injury. PMID- 10788264 TI - Great regulatory expectations. AB - In a remarkably short time, the European regulatory landscape for medical devices has changed dramatically and permanently. The speed of this change will continue to accelerate until a fully matured body of rules is in place. This article will examine the current status of regulations in Europe, and some areas that will need to be addressed in the next century. PMID- 10788265 TI - The 1999 contract services survey. PMID- 10788266 TI - The 1999 Medical Device Technology Employment Survey. PMID- 10788267 TI - Germany dominates. Espicom Ltd. PMID- 10788268 TI - France cutting costs. Espicom Ltd. PMID- 10788269 TI - More requirements for doing business in France. PMID- 10788270 TI - Italy. Failure to invest. Espicom Ltd. PMID- 10788271 TI - The United Kingdom. A mature market. Espicom Ltd. PMID- 10788273 TI - The Netherlands multinational centre. PMID- 10788272 TI - Denmark high spending. PMID- 10788274 TI - Sweden open to innovation. Espicom Ltd. PMID- 10788275 TI - Finland. The great exporter. Espicom Ltd. PMID- 10788276 TI - Belgium active distribution centre. Espicom Ltd. PMID- 10788277 TI - Greece raising standards. Espicom Ltd. PMID- 10788278 TI - New technologies for the 21st century. AB - In the 21st century, medical science will move beyond the practice of transplantation and into fabrication of artificial organs using tissue and generative engineering. Artificial organs will be used not only to save lives at the end stage of diseases, but also to support daily life and to prevent disease. This article reviews some of the microelectromechanical systems that are being developed, which are key technologies for future artificial organs. PMID- 10788279 TI - Denial is not a qualitative concept. PMID- 10788280 TI - Analysis on the move: deconstructing troublesome health questions and troubling epidemiology. AB - Qualitative research gets close to experiences of pain, illness, and disease; consequently, qualitative researchers often find themselves asked troublesome questions (i.e., laypeople ask for practical, helpful answers to their everyday illness concerns). This is not surprising, but of interest is the fact that academics ask each other such troublesome questions as part of academic discourse. When academics ask such questions, they may sometimes be after practical information, but they may also be using the questioning as an attack on the supposed excessive relativism of social constructionism. Three key analytical moves that offer a useful deconstruction of troublesome health questions are outlined, showing that they are another useful topic of constructionist inquiry. To lessen abstraction, these moves are brought to bear on a case study of a possible connection between pesticide use and birth defects, thus showing how social science and epidemiology can be connected, troubled, and extended in the process. PMID- 10788281 TI - Self-harm: cutting the bad out of me. AB - The practice of self-harm is increasing in the United Kingdom, accounting for the highest number of acute medical admissions for women. The medical and nursing response to repeaters, set within a climate of dwindling emergency and accident resources, has been one of impatience, frustration, and hostile care. The author undertook a correspondence study with 6 women who regularly self-harmed. The women claimed that medical and nursing professionals viewed their self-harm as irrational and illogical. However, a qualitative examination of the motivations and interests of all parties reveals that self-harm acts possess situated internal logic, whereas professionals tend to use rational logic in attempting to understand them. PMID- 10788282 TI - Hoping to maintain a balance: the concept of hope and the discontinuation of anorexiant medications. AB - The secondary analysis of transcripts from a study of 9 women who used prescription anorexiant medications described in this article was conducted to discover the role that hope played in the women's weight-loss initiatives. It was found that hope grew throughout the women's weight-loss initiative but peaked once the medications were discontinued. Seven components of the concept of hope were described in the context of the women's weight-loss experiences. Characteristics of hope in women who have discontinued anorexiant medications are different than those associated with hope in other contexts. A unique pattern of hope, hoping to maintain a balance, describes the women's experiences as they strove to maintain the weight they had lost. PMID- 10788283 TI - Aboriginal grandmothers' experience with health promotion and participatory action research. AB - This article describes a case study examining the effects of participating in a health promotion project, one aspect of which was a health assessment conducted using participatory action research. The study was carried out over 2.5 years in a project for older Aboriginal women (hereafter known as the grandmothers). Participation in the project and health assessment contributed to a number of changes in them, which were categorized as cleansing and healing, connecting with self, acquiring knowledge and skills, connecting within the group, and external exposure and engagement. This experience demonstrated an approach to health promotion programming and conducting a health assessment that was acceptable to this group of people and fostered changes congruent with empowerment. PMID- 10788284 TI - Creative response to alternative medicine: clients of a modern Finnish healer in a northwestern Ontario city. AB - Antti Auttaja is a Finnish Canadian businessman who practices alternative medicine in a northwestern Ontario city. His healing method involves pressing his hands and fingers firmly but gently on the feet, wrists, stomach, chest, and other bodily areas of his clients, who recline on their backs on the floor, with pillows propped under their head and knees. His clients respect him and his healing ability, and as the questionnaire and interview data presented in this article clearly show, they feel he has helped them enhance their quality of life in a way that is unique to him and very special to them. He exemplifies a holistic healer working along the periphery of the modern biomedical system. His practice exemplifies the alternative medicine category of the domain of nonconventional medicine. PMID- 10788285 TI - Organizational characteristics and issues affecting the longevity of self-help groups for parents of children with special needs. AB - In this article, the organizational characteristics and issues affecting the maintenance of nine self-help groups for parents of children with special needs are discussed. Qualitative data were collected through interviews with 20 parents and observation of six group meetings. For continued existence, groups must deal with the challenges of encouraging new leaders, attracting new members, obtaining funds or assistance to support their activities, and meeting the changing needs of members. Important organizational characteristics associated with longevity are committed and effective leadership, community connections that help provide needed funds or practical assistance, and the willingness of group members to change activities to meet changing needs. This information can be used by parents and service providers to plan new groups, recruit group members, and ensure the effective functioning of groups as they change over time. PMID- 10788286 TI - Predictive genetic testing in diabetes: a case study of multiple perspectives. AB - Genetic testing is now possible in maturity onset diabetes of the young (MODY), an unusual genetic subtype of diabetes. To date, there has been no research into the implications of genetic testing for MODY families. The case study of the first known instance of predictive genetic testing for diabetes described in this article focuses on the perspectives of the family and the health care professionals involved in the decision-making process. Open-ended interviews were conducted before and after the predictive test. Content analysis highlighted four key areas: autobiographical experiences, motivations for testing, competing priorities in genetic counseling, and differing attitudes to predictive testing for children. Reactions to the predictive test result are presented. The implications of these findings for the provision of predictive genetic testing in diabetes are considered. PMID- 10788287 TI - The peer advisor experience providing social support. AB - Research shows a link between increased social support and decreased complications after myocardial infarction (MI). In a current randomized controlled trial (RCT), a social support intervention administered by nurse and peer advisors is being examined to determine its influence on the health outcomes of unpartnered, post-MI elders. This qualitative study (as part of the larger RCT) sought a better understanding regarding the experience of the peer advisor. Data sources included peer advisor logs, a focus group, and telephone interviews with peer advisors. Findings revealed that helping, mutual sharing, committing, and benefiting are characteristics of peer experiences. Primarily because of their personal experience of recovery from MI, peer advisors had a remarkable ability to relate to assigned post-MI elders, offering a unique form of social support complementary to current health practices. PMID- 10788288 TI - Putting molecular methods to work. PMID- 10788289 TI - Hematology makes the shift to the core lab. PMID- 10788290 TI - How to work up hypercoagulability. PMID- 10788291 TI - Hematology analyzers. PMID- 10788292 TI - Teasing out the value of new C-reactive protein test. PMID- 10788293 TI - Tennessee court finds liability without negligence. PMID- 10788294 TI - Digital imaging not picture perfect--yet. PMID- 10788295 TI - Layers of 'ifs' bedevil antiangiogenesis. PMID- 10788296 TI - Using new D-dimer tests to rule out venous thromboembolism. PMID- 10788298 TI - CLA hones systems, standards, questions, directions. PMID- 10788297 TI - Weeding out the wrong coagulation analyzer. PMID- 10788299 TI - Unveiling the patterns in TDM data 'dots'. PMID- 10788301 TI - What Indianapolis ERs are learning by linking. PMID- 10788300 TI - Upping the ante in biochip research. PMID- 10788302 TI - Lab discounts. How deep is too deep? PMID- 10788303 TI - Marching toward single-donor NAT. PMID- 10788304 TI - Mothers' responses to sons and daughters engaging in injury-risk behaviors on a playground: implications for sex differences in injury rates. AB - Videotapes of children engaging in injury-risk activities on a playground were shown to mothers, who were asked to intervene by stopping the tape and saying whatever they would to their child in the situation shown. Results revealed that mothers of daughters were more likely to judge behaviors as posing some degree of injury risk, and they intervened more frequently and quickly than mothers of sons. Mothers' speed to intervene positively correlated with both children's injury history and their risk-taking tendencies, indicating that mothers of children who were previously injured and who often engaged in injury-risk behaviors had a higher degree of tolerance for children's risk taking than mothers of children who experienced fewer injuries and less frequently engaged in injury-risk behaviors. Mothers' verbalizations to children's risk taking revealed that daughters received more cautions and statements communicating vulnerability for injury, whereas sons received more statements encouraging risk-taking behavior. PMID- 10788305 TI - The development of automaticity in accessing number magnitude. AB - This study traces developmental changes in automatic and intentional processing of Arabic numerals using a numerical-Stroop paradigm. In Study 1, university students compared the numerical or physical size of Arabic numerals varying along both dimensions. In Study 2, first graders (mean age = 6 years 6 months), third graders (mean age = 8 years 4 months), and fifth graders (mean age = 10 years 3 months) were tested to examine developmental changes in numerical and physical comparisons. In the numerical comparison task, a size congruity effect was found at all ages (i.e., relative to a neutral control, congruent physical sizes facilitated, and incongruent sizes interfered with, the numerical comparison). The pattern of facilitation and interference, however, was modulated by age. In the physical comparison task, the incongruity between physical and numerical size affected only older children and adults. These findings strongly suggest that the automatization in number processing is achieved gradually as numerical skills progress. PMID- 10788306 TI - Developing normal reading skills: aspects of the visual processes underlying word recognition. AB - Visual word recognition performance of first graders (mean age: 6.6 years) through fifth graders (mean age: 10.8 years) was investigated using an experimental technique that is known to elicit the "viewing position effect" in skilled readers. The results showed that this effect, which consists of a systematic variation of performance as a function of fixation position within words, emerged early at the end of the 1st year of reading instruction. Visual field asymmetries in recognizing individual letters in words were also observed starting from first grade. Effects of word familiarity were obtained as early as in second grade. In contrast to skilled readers, children showed a marked word length effect, which persisted through the first 5 years of instruction. No other qualitative differences between beginning and skilled readers were apparent. Hence, the basics of reading skills, as measured by the present technique, seem to be attained very early during acquisition. Further experience mainly reduces the time a reader needs to extract visual information from print. PMID- 10788307 TI - Persistence of memory for ignored lists of digits: areas of developmental constancy and change. AB - Contrary to the common belief that sensory memory remains unchanged across development in childhood, there have been several previous reports suggesting that the persistence of sensory memory, at least for sounds, increases with age in childhood. Because those previous studies all used isolated sounds as stimuli, it is not yet clear how this developmental difference influences the recall of sound series. The present study adapts the procedure of J. S. Saults and N. Cowan (1996), who studied memory for attended and ignored spoken words, to examine here the recall of attended and ignored lists of digit. A developmental increase in the persistence of memory was obtained only for the final item in an ignored list, which is the item for which sensory memory is thought to be the most vivid at short retention intervals. PMID- 10788308 TI - Mitochondrial genome-encoded ATPase subunit 6+8 mRNA increases in human hepatoblastoma cells in response to nonfatal cold stress. AB - Cellular responses to cold stress have not been well clarified, compared with heat shock responses, especially in mammalian cells. We investigated cold-stress responses in human hepatoblastoma cells (HepG2) exposed to a nonfatal temperature of 17 degrees C. Under the condition, RNA and protein syntheses in the cells were highly, but incompletely, depressed and cell growth was impaired. A cDNA subtraction method was used to isolate mRNAs for which the levels were increased in cold-stressed cells compared with cells cultured at 37 degrees C. A transcript isolated by the screening was identified as ATPase subunit 6+8 mRNA that encodes components of a mitochondrial ATPase complex and that is transcribed from a mitochondrial genome. The copy number of the mitochondrial genome in cells was not changed by cold stress. Thus, HepG2 cells were treated with various concentrations of actinomycin D and chloramphenicol to assess the effects of transcriptional and translational reduction on the increased level of the ATPase subunit 6+8 mRNA. The mRNA level was increased in cells treated with low concentrations of the RNA or protein synthesis inhibitors. These results indicate that the increase in ATPase subunit 6+8 mRNA stimulated by cold stress could be mediated by a partial decline of transcription and/or translation in the cells. In addition, the degradation of ATPase subunit 6+8 mRNA was suppressed in cold stressed cells compared with that in 37 degrees C-cultured cells. This result implies that posttranscriptional regulation is also involved in the cold stimulated increase in ATPase subunit 6+8 mRNA in HepG2 cells. PMID- 10788309 TI - Cryobiology of articular cartilage: ice morphology and recovery of chondrocytes. AB - The cryopreservation of articular cartilage chondrocytes has been achieved with cells isolated from the cartilage matrix but has found only limited success when the tissue is left intact. Previous work with ovine cartilage has shown that cryopreservation of the chondrocytes of the superficial and deep zones is possible, but the cells of the intermediate zone have not been successfully cryopreserved. This finding led to the suggestion that there might be biological differences between chondrocytes of the different morphological zones that were responsible for this differential recovery. This study investigates the hypothesis that the cells of the intermediate zone are more sensitive to cryoinjury by introducing cuts in the cartilage so that cells of the intermediate zone have the same proximity to the outer surface of the tissue as the cells of the superficial zone. When this was done, it was found that cells of the intermediate zone could survive cryopreservation as well as the cells of the superficial zone when they were near a surface, but not when they were embedded deep within the tissue. Thus the hypothesis of a biological difference between the cells of the two zones being responsible for the differential recovery is disproved. It is further hypothesized that physical proximity to a surface leads to higher recovery as a result of planar ice growth into the cartilage. PMID- 10788310 TI - Viral contamination of embryos cryopreserved in liquid nitrogen. AB - Despite the worldwide application of embryo-freezing technology as the means of preserving germplasm of mammalian species, there is no information available on the possible transmission of infectious agents to cryopreserved embryos via contaminated liquid nitrogen (LN). Recently, it has been reported that new methods of cryopreservation which employ ultrarapid freezing or vitrification require direct contact between the freezing medium containing oocytes or embryos and liquid phase nitrogen (LPN). As models for human and animal viral pathogens three bovine viruses, bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV), and bovine immunodeficiency virus (BIV), were employed to study the potential for their transmission by experimentally contaminated LN to embryos frozen and stored in open freezing containers. Bovine embryos in a mixture of 20% ethylene glycol, 20% ME(2)SO, and 0.6% sucrose were vitrified in either unsealed standard 0.25 ml or modified open pulled straws or in plastic cryovials and then plunged into contaminated LPN. After 3-5 weeks of storage in LN, embryos were thawed and sequentially washed and only those with intact ZP were pooled together and tested in batches of three for viral contamination. From this pool of 83 batches, 13 of 61 (21.3%) batches exposed to BVDV and BHV-1 tested positive for viral association while all 22 batches exposed to BIV in unsealed containers tested negative. All control embryos vitrified in sealed cryovials and straws were free from viral contamination. PMID- 10788311 TI - Development of a simple sperm cryopreservation model using a chemically defined medium and goat cauda epididymal spermatozoa. AB - This investigation was carried out to develop a simple sperm cryopreservation model using a chemically defined synthetic medium (modified Ringer's solution) and mature goat cauda epididymal sperm as the model system. Rates of cooling, freezing, and maximum freezing temperature were manipulated with the help of a computer-controlled programmable biofreezer. Highly motile goat cauda sperm dispersed in a modified Ringer's solution was subjected to the freezing protocol: cooling 0.25 degrees C min(-1) to 5 degrees C, 5 degrees C min (-1) to -20 degrees C, 20 degrees C min(-1) to -100 degrees C, prior to plunging into liquid nitrogen. In the absence of any cryoprotective agent, all of the spermatozoa lost their motility. Addition of glycerol (0.22 to 0.87 M) caused a dose-dependent increase of sperm motility recovery. The highest recovery of forward and total motility was (32 and 35%, respectively) at 0.87 M. Further increase of the glycerol concentration caused a marked decrease in motility. Changes in the cooling rate particularly before and during freezing had a notable effect on the sperm motility recovery. There was no or low recovery (0-18%) of sperm motility when the cells were transferred directly to liquid nitrogen from the initial two cooling stages. The data demonstrate the importance of all of the cooling stages in the cryopreservation of the cells. Like glycerol, dimethyl sulfoxide (Me(2)SO) and ethylene glycol also showed a dose-dependent increase in motility recovery as well as a biphasic curve of cryoprotection. At optimal concentrations, dimethyl sulfoxide (1.00 M) and ethylene glycol (1.29 M) were effective in recovering sperm motility to the extent of 20 and 13%, respectively. Thus these reagents have markedly lower cryoprotection potential than glycerol. PMID- 10788312 TI - Electromagnetic rewarming: the effect of CPA concentration and radio source frequency on uniformity and efficiency of heating. AB - The relevant dielectric properties of a practical perfusate formula have been measured at radiofrequencies from 27 to 2954 MHz and temperatures from -75 to +25 degrees C. Both dimethyl sulfoxide and 2,3-butanediol were investigated as the cryoprotectant component at concentrations from 15 to 50%. The dielectric properties are considered in terms of new indices which characterize uniformity of heating, the power delivered and the efficiency of heating, and the avoidance of localized thermal runaway during very rapid heating. It becomes clear that a compromise must be accepted between nonuniformity of heating at the higher frequencies and inconveniently low power delivery at the lower frequencies. The choice of CPA and the concentration are found to be not very significant in these respects. PMID- 10788314 TI - Glass-forming tendency in the system water-dimethyl sulfoxide. AB - The glass-forming tendency on cooling and the stability of the wholly amorphous state on warming have been previously reported for many cryoprotective solutions. However, unlike the other solutions, those of dimethyl sulfoxide (Me(2)SO) have not been studied on cooling. In this paper, the glass-forming tendency of Me(2)SO aqueous solutions has been measured for solutions containing 40, 43, 45, and 47.5% (w/w) Me(2)SO. At a concentration of 45% (w/w), the glass-forming tendency decreases in the following order: levo-2, 3-butanediol, 1,3-butanediol, 1,2 propanediol, 1,2,3-butanetriol, dimethyl sulfoxide, dimethylformamide, diethylformamide, 1, 4-butanediol, ethylene glycol, glycerol, 1,3-propanediol. New measurements have also been made on warming the Me(2)SO solutions. PMID- 10788313 TI - Osmotic effects on feline spermatozoa from normospermic versus teratospermic donors. AB - Effects of osmolality stresses on the sperm of normospermic (>60% normal sperm/ejaculate) versus teratospermic (<40% normal sperm) domestic cats and the normospermic leopard cat and the teratospermic clouded leopard were studied. Spermatozoa were exposed to various anisotonic solutions in a single step or returned to near isotonic conditions in a single step after exposure to anisotonic solutions. The percentage of sperm motility was measured subjectively, and dual fluorescent stains were used to assess membrane integrity by flow cytometry. The percentage of sperm motility declined (P < 0.05) in domestic cat sperm exposed to osmolalities <200 and >450 mOsm. Spermatozoa from all felines underwent marked (P < 0.05) membrane disruption following a hypotonic stress, but sperm from teratospermic donors experienced greater (P < 0.05) membrane disruption in response to decreased osmolality. While feline spermatozoa appeared to be highly resistant to hypertonic (600, 1200, and 2400 mOsm) conditions, with >85% of the cells maintaining intact membranes, severe membrane disruption occurred when cells were returned to isotonicity in a single step. There was no difference (P > 0.05) between a 1- and 5-min exposure to various anisotonic solutions. Similarly, sperm from normospermic and teratospermic domestic cats responded identically after exposure to ionic or nonionic solute. Results demonstrate that: (1) spermatozoa from teratospermic males are more vulnerable to a hypotonic stress than sperm from normospermic counterparts; (2) in response to small deviations in osmolality, feline sperm experience a more rapid decline in motility than membrane integrity; and (3) an abrupt return to isotonicity after a hypertonic stress causes extensive sperm membrane damage regardless of ejaculate quality. PMID- 10788315 TI - An in vitro monitoring system for simulated thermal process in cryosurgery. AB - This paper describes a new in vitro experimental model that records temperature changes over a culture plate, which then can be used to assess the biological effects of cryosurgery. A cryoprobe and 16 thermocouples set up by a computer control system were used to monitor the temperature changes during freezing and thawing in a culture plate, and the data were used to create a temperature profile of the entire plate. Location of the thermocouples was confirmed by a digital camera viewing from under the plate, and temperature changes at any point in the interpolated areas were estimated using a curve fitting method. The estimated temperature was checked by sampling with four additional randomly placed thermocouples. Linear regression analysis showed that the estimated temperature and measured temperature were very close (correlation coefficients 0.98-0.99). MBT-2 tumor cells were cultured and then subjected to simulated cryosurgery. The surviving cells were stained with crystal violet and the cell death boundary was detected by image processing. Temperature history at the cell death boundary was retrieved and analyzed. With this system it is possible to recreate the temperature changes that result in a certain biological effect (such as cell death) during the process of simulated cryosurgery. PMID- 10788316 TI - Expression of mRNAs of the aquaporin family in mouse oocytes and embryos. AB - The molecular basis of water and cryoprotectant permeability in mammalian oocytes and embryos is poorly understood. Therefore, we investigated the expression of mRNAs of water channel proteins (aquaporins) in mouse oocytes and embryos by RT PCR. The total RNA of mouse oocytes at metaphase II and embryos at the 4-cell, morula, and blastocyst stages was isolated, reverse-transcribed, and subjected to nested PCR amplification. Aquaporins were expressed in both oocytes and embryos, but the types were different among the developmental stages: aquaporins 3 and 7 were expressed in oocytes and embryos at all stages examined, but aquaporins 8 and 9 were expressed only in blastocysts. On the other hand, aquaporins 1, 2, 4, 5, and 6 were not detected in any of the stages examined. The present study shows for the first time that aquaporins are expressed in mammalian oocytes and embryos. These aquaporins may play a role in water transport and conceivably also in cryoprotectant transport across the plasma membrane in these cells. PMID- 10788317 TI - Ultrastructural changes in hypothermically preserved hepatocytes. AB - Hypothermic preservation of hepatocytes on gelatin gels (10 degrees C) provided a stable system for investigating cold-induced changes culminating in cell death. Hepatocyte morphology remained unchanged during 9 days of preservation. Thereafter there was a progressive movement of organelles toward the center of the cell. During this process the mitochondria appeared to have a normal morphology suggesting that they are not the primary cause of the transition. When the movement was completed the mitochondria appeared aggregated and microvilli were no longer apparent on the cell surface. PMID- 10788318 TI - Gaseous oxygen for hypothermic preservation of predamaged liver grafts: fuel to cellular homeostasis or radical tissue alteration? AB - Gaseous persufflation with molecular oxygen during cold storage has recently been discussed as an adjunct in the preservation of predamaged livers from non-heart beating donors. In this study, the actual importance of an antioxidant medication prior to oxygen persufflation was investigated for the first time. Rat livers were excised 60 min after cardiac arrest of the donor, flushed with UW, and stored at 4 degrees C for 24 h. Gaseous oxygen, applied to the livers during the storage period via the caval vein at a pressure limited to 18 mm Hg, was only effective in improving hepatic viability upon reperfusion when antioxidants (superoxide dismutase (SOD) or n-acetylcysteine) were added to the UW rinse solution. Either drug prevented an increase in free radical mediated lipid peroxidation. Acetylcysteine but not SOD also counteracted the phosphorylation of Ikappab and thus the transcriptional activation of NFkappab. Poor results after oxygen persufflation without antioxidants may hence be explained by concomitant free radical induced tissue alterations upon introduction of molecular oxygen into ischemically predamaged organs. PMID- 10788319 TI - Effects of phosphorothioate modifications on precursor tRNA processing by eukaryotic RNase P enzymes. AB - The cleavage mechanism has been studied for nuclear RNase P from Saccharomyces cerevisiae, Homo sapiens sapiens and Dictyostelium discoideum, representing distantly related branches of the Eukarya. This was accomplished by using precursor tRNAs (ptRNAs) carrying a single Rp or Sp-phosphorothioate modification at the normal RNase P cleavage site (position -1/+1). All three eukaryotic RNase P enzymes cleaved the Sp-diastereomeric ptRNA exclusively one nucleotide upstream (position -2/-1) of the modified canonical cleavage site. Rp-diastereomeric ptRNA was cleaved with low efficiency at the modified -1/+1 site by human RNase P, at both the -2/-1 and -1/+1 site by yeast RNase P, and exclusively at the -2/-1 site by D. discoideum RNase P. The presence of Mn(2+ )and particularly Cd(2+) inhibited the activity of all three enzymes. Nevertheless, a Mn(2+ )rescue of cleavage at the modified -1/+1 site was observed with yeast RNase P and the Rp diastereomeric ptRNA, consistent with direct metal ion coordination to the (pro) Rp substituent during catalysis as observed for bacterial RNase P enzymes. In summary, our results have revealed common active-site constraints for eukaryotic and bacterial RNase P enzymes. In all cases, an Rp as well as an Sp phosphorothioate modification at the RNase P cleavage site strongly interfered with the catalytic process, whereas substantial functional interference is essentially restricted to one of the two diastereomers in other RNA and protein catalyzed hydrolysis reactions, such as those catalyzed by the Tetrahymena ribozyme and nuclease P1. PMID- 10788320 TI - The human homologue of the yeast splicing factor prp6p contains multiple TPR elements and is stably associated with the U5 snRNP via protein-protein interactions. AB - An essential step of pre-mRNA spliceosome assembly is the interaction between the snRNPs U4/U6 and U5, to form the [U4/U6.U5] tri-snRNP. While the tri-snRNP protein Prp6p appears to play an important role for tri-snRNP formation in yeast, little is known about the interactions that connect the two snRNP particles in human tri-snRNPs. Here, we describe the molecular characterisation of a 102kD protein form HeLa tri-snRNPs. The 102kD protein exhibits a significant degree of overall homology with the yeast Prp6p, including the conservation of multiple tetratrico peptide repeats (TPR), making this the likely functional homologue of Prp6p. However, while the yeast Prp6p is considered to be a U4/U6-specific protein, the human 102kD protein was found to be tightly associated with purified 20 S U5 snRNPs. This association appears to be primarily due to protein-protein interactions. Interestingly, antibodies directed against the C-terminal TPR elements of the 102kD protein specifically and exclusively immunoprecipitate free U5 snRNPs, but not [U4/U6.U5] tri-snRNPs, from HeLa nuclear extract, suggesting that the C-terminal region of the 102kD protein is covered by U4/U6 or tri-snRNP specific proteins. Since proteins containing TPR elements are typically involved in multiple protein-protein interactions, we suggest that the 102kD protein interacts within the tri-snRNP with both the U5 and U4/U6 snRNPs, thus bridging the two particles. Consistent with this idea, we show that in vitro translated U5 102kD protein binds to purified 13S U4/U6 snRNPs, which contain, in addition to the Sm proteins, all known U4/U6-specific proteins. PMID- 10788321 TI - Overproduced Salmonella typhimurium flagellar motor switch complexes. AB - Three Salmonella typhimurium flagellar motor proteins, FliG, FliM and FliN, are required for the switching of rotation sense. The proteins have been localized to the cytoplasmic module of the flagellar base. Structures, which were morphologically indistinguishable from the native transmembrane MS-ring and cytoplasmic C-ring basal body modules, formed in Escherichia coli upon plasmid encoded synthesis of these proteins together with FliF. The structures localized to the cell membrane and contained all three motor proteins, as determined by immuno-electron microscopy. This result supports the deduction, based on earlier biochemical analysis, that the C-ring is composed entirely of these proteins and, therefore, functions as a dedicated motor component. In addition, it demonstrates that the morphologically correct assembly of the C-ring onto the MS-ring proceeds independently of other structural components of these modules. PMID- 10788322 TI - The analysis of the transcriptional activator PrnA reveals a tripartite nuclear localisation sequence. AB - Nuclear localisation signals (NLSs) have been classified as either mono- or bipartite. Genetic analysis and GFP fusions show that the NLS of a Zn-binuclear cluster transcriptional activator of Aspergillus nidulans (PrnA) is tripartite. This NLS comprises two amino-terminal basic sequences and the first basic sequence of the Zn-cluster. Neither the two amino-terminal basic sequences nor the paradigmatic nucleoplasmin bipartite NLS drive our construction to the nucleus. Cryosensitive mutations in the second basic sequence are suppressed by mutations that restore the basicity of the domain. The integrity of the Zn cluster is not necessary for nuclear localisation. A tandem repetition of the two basic amino-terminal sequences results in a strong NLS. Complete nuclear localisation is observed when the whole DNA-binding domain, including the putative dimerisation element, is included in the construction. At variance with what is seen with tandem NLSs, all fluorescence here is intra-nuclear. This suggests that retention and nuclear entry are functionally different. With the whole PrnA protein, we observe localisation, retention and also a striking sub localisation within the nucleus. Nuclear localisation and sub-localisation are constitutive (not dependent on proline induction). In contrast with what has been observed by others in A. nidulans, none of our constructions are delocalised during mitosis. This is the first analysis of the NLS of a Zn-binuclear cluster protein and the first characterisation of a tripartite NLS. PMID- 10788323 TI - Binding of EcoP15I DNA methyltransferase to DNA reveals a large structural distortion within the recognition sequence. AB - EcoP15I DNA methyltransferase, a member of the type III restriction-modification system, binds to the sequence 5'-CAGCAG-3' transferring a methyl group from S adenosyl-l-methionine to the second adenine base. We have investigated protein DNA interactions in the methylase-DNA complex by three methods. Determination of equilibrium dissociation constants indicated that the enzyme had higher affinity for DNA containing mismatches at the target base within the recognition sequence. Potassium permanganate footprinting studies revealed that there was a hyper reactive permanganate cleavage site coincident with adenine that is the target base for methylation. More importantly, to detect DNA conformational alterations within the enzyme-DNA complexes, we have used a fluorescence-based assay. When EcoP15I DNA methyltransferase bound to DNA containing 2-aminopurine substitutions within the cognate sequence, an eight to tenfold fluorescent enhancement resulting from enzymatic flipping of the target adenine base was observed. Furthermore, fluorescence spectroscopy analysis showed that the changes attributable to structural distortion were specific for only the bases within the recognition sequence. More importantly, we observed that both the adenine bases in the recognition site appear to be structurally distorted to the same extent. While the target adenine base is probably flipped out of the DNA duplex, our results also suggest that fluorescent enhancements could be derived from protein DNA interactions other than base flipping. Taken together, our results support the proposed base flipping mechanism for adenine methyltransferases. PMID- 10788324 TI - Methyl-specific DNA binding by McrBC, a modification-dependent restriction enzyme. AB - McrBC, a GTP-requiring, modification-dependent endonuclease of Escherichia coli K 12, specifically recognizes DNA sites of the form 5' R(m)C 3'. DNA cleavage normally requires translocation-mediated coordination between two such recognition elements at distinct sites. We have investigated assembly of the cleavage-competent complex with gel-shift and DNase I footprint analysis. In the gel-shift system, McrB(L) binding resulted in a fast-migrating specific shifted band, in a manner requiring both GTP and Mg(2+). The binding was specific for methylated DNA and responded to local sequence changes in the same way that cleavage does. Single-stranded DNA competed for McrB(L)-binding in a modification and sequence-specific fashion. A supershifted species was formed in the presence of McrC and GTPgammaS. DNase I footprint analysis showed modest cooperativity in binding to two sites, and a two-site substrate displayed protection in non specific spacer DNA in addition to the recognition elements. The addition of McrC did not affect the footprint obtained. We propose that McrC effects a conformational change in the complex rather than a reorganization of the DNA:protein interface. PMID- 10788325 TI - Altering molecular recognition of RNA aptamers by allosteric selection. AB - In a continuing effort to explore structural and functional dynamics in RNA catalysis, we have created a series of allosteric hammerhead ribozymes that are activated by theophylline. Representative ribozymes exhibit greater than 3000 fold activation upon effector-binding and cleave with maximum rate constants that are equivalent to the unmodified hammerhead ribozyme. In addition, we have evolved a variant allosteric ribozyme that exhibits an effector specificity change from theophylline to 3-methylxanthine. Molecular discrimination between the two effectors appears to be mediated by subtle conformational differences that originate from displacement of the phosphodiester backbone near the effector binding pocket. These findings reveal the importance of abstruse aspects of molecular recognition by nucleic acids that are likely to be unapproachable by current methods of rational design. PMID- 10788326 TI - Reassembly of Lumbricus terrestris hemoglobin: a study by matrix-assisted laser desorption/ionization mass spectrometry and 3D reconstruction from frozen hydrated specimens. AB - Dodecamers and four types of linker chains (L1-L4) were purified from dissociated hemoglobin of the earthworm Lumbricus terrestris. Various preparations comprising dodecamer of globin chains and linker chains were allowed to reassemble at neutral pH. They produced various oligomers that were purified by gel filtration, analyzed in matrix-assisted laser desorption/ionization mass spectrometry and submitted to 3D reconstruction from isolated particles observed in cryoelectron microscopy. Despite the impossibility to completely free the L2, L3, and L4 preparations from L1, the following conclusions were obtained. First, hemoglobin molecules indistinguishable from native hemoglobin at 25 A resolution were obtained in the absence of linker chains L2, L3, or L4. Second, the 3D reconstruction volumes of reassembled hemoglobins containing dodecamers and L1+L3 or dodecamers and L1+L4 demonstrate that reassembly of native-like structures can be obtained from at most two linker chains and dodecamers. Third, the 3D reconstruction volumes of native and reassembled hemoglobins containing dodecamers and (1) L1, L2, and L4, (2) L1, L3, and L4, (3) L1 and L4, and (4) L1 and L3 were highly similar. Since these structures comprise two types of substructures (one involved in the c3a, c3b, and c4 linking units of the hollow globular substructure and the other in the c5 connection and the toroid), it seems highly probable that the minimal number of linker chains required to reassemble native-like hemoglobin is at most two. PMID- 10788327 TI - Uncoupling actin filament fragmentation by cofilin from increased subunit turnover. AB - The actin depolymerizing factor (ADF)/cofilin family of proteins interact with actin monomers and filaments in a pH-sensitive manner. When ADF/cofilin binds F actin it induces a change in the helical twist and fragmentation; it also accelerates the dissociation of subunits from the pointed ends of filaments, thereby increasing treadmilling or depolymerization. Using site-directed mutagenesis we characterized the two actin-binding sites on human cofilin. One target site was chosen because we previously showed that the villin head piece competes with ADF for binding to F-actin. Limited sequence homology between ADF/cofilin and the part of the villin headpiece essential for actin binding suggested an actin-binding site on cofilin involving a structural loop at the opposite end of the molecule to the alpha-helix already implicated in actin binding. Binding through the alpha-helix is primarily to monomeric actin, whereas the loop region is specifically involved in filament association. We have characterized the actin binding properties of each site independently of the other. Mutation of a single lysine residue in the loop region abolishes binding to filaments, but not to monomers. Using the mutation analogous to the phosphorylated form of cofilin (S3D), we show that filament binding is inhibited at physiological ionic strength but not under low salt conditions. At low ionic strength, this mutant induces both the twist change and fragmentation characteristic of wild-type cofilin, but does not activate subunit dissociation. The results suggest a two-site binding to filaments, initiated by association through the loop site, followed by interaction with the adjacent subunit through the "helix" site at the opposite end of the molecule. Together, these interactions induce twist and fragmentation of filaments, but the twist change itself is not responsible for the enhanced rate of actin subunit release from filaments. PMID- 10788328 TI - Microtubules switch occasionally into unfavorable configurations during elongation. AB - Tubulin assembles to form a range of structures that differ by their protofilament and monomer helix-start numbers. The microtubule lattice is believed to accommodate these different configurations by skewing the protofilaments so that the lateral interactions between tubulin subunits are maintained. Here, we present the characterization of 14 types of microtubules, including six novel ones, through an extensive analysis of microtubules assembled in vitro from pure tubulin. Although the six new types represented only 1 % of the total length of the population examined ( approximately 17 mm), they define the limits of microtubule structure and assembly. Protofilament skewing is restricted to within +/-2 degrees. Outside this range, the restoring force induced by the skewed protofilaments is compensated by a longitudinal shift (less than +/-0.2 nm) between adjacent protofilaments. Configurations with theoretical protofilament skew angles larger than +/-4 degrees or that necessitate larger modifications of the microtubule surface lattice were not observed. Analysis of the microtubule types distribution reveals that it is sharply peaked around the less skewed conformations. These results indicate that both the flexibility of the protofilaments and the strength of their lateral interactions restrict the range of structures assembled. They also demonstrate that growing microtubules can occasionally switch into energetically unfavorable configurations, a behavior that may account for the stochastic nature of catastrophes. PMID- 10788329 TI - Anomalous diffusion of fluorescent probes inside living cell nuclei investigated by spatially-resolved fluorescence correlation spectroscopy. AB - We have investigated spatial variations of the diffusion behavior of the green fluorescent protein mutant EGFP (F64L/S65T) and of the EGFP-beta-galactosidase fusion protein in living cells with fluorescence correlation spectroscopy. Our fluorescence correlation spectroscopy device, in connection with a precision x-y translation stage, provides submicron spatial resolution and a detection volume smaller than a femtoliter. The fluorescence fluctuations in cell lines expressing EGFP are caused by molecular diffusion as well as a possible internal and a pH dependent external protonation process of the EGFP chromophore. The latter processes result in two apparent nonfluorescent states that have to be taken into account when evaluating the fluorescence correlation spectroscopy data. The diffusional contribution deviates from ideal behavior and depends on the position in the cell. The fluorescence correlation spectroscopy data can either be evaluated as a two component model with one fraction of the molecules undergoing free Brownian motion with a diffusion coefficient approximately five times smaller than in aqueous solution, and another fraction diffusing one or two orders of magnitude slower. This latter component is especially noticeable in the nuclei. Alternatively, we can fit the data to an anomalous diffusion model where the time dependence of the diffusion serves as a measure for the degree of obstruction, which is large especially in nuclei. Possible mechanisms for this long tail behavior include corralling, immobile obstacles, and binding with a broad distribution of binding affinities. The results are consistent with recent numerical models of the chromosome territory structure in the cell nucleus. PMID- 10788330 TI - The crystal structure of a sulfurtransferase from Azotobacter vinelandii highlights the evolutionary relationship between the rhodanese and phosphatase enzyme families. AB - Rhodanese is an ubiquitous enzyme that in vitro catalyses the transfer of a sulfur atom from suitable donors to nucleophilic acceptors by way of a double displacement mechanism. During the catalytic process the enzyme cycles between a sulfur-free and a persulfide-containing form, via formation of a persulfide linkage to a catalytic Cys residue. In the nitrogen-fixing bacteria Azotobacter vinelandii the rhdA gene has been identified and the encoded protein functionally characterized as a rhodanese. The crystal structure of the A. vinelandii rhodanese has been determined and refined at 1.8 A resolution in the sulfur-free and persulfide-containing forms. Conservation of the overall three-dimensional fold of bovine rhodanese is observed, with substantial modifications of the protein structure in the proximity of the catalytic residue Cys230. Remarkably, the native enzyme is found as the Cys230-persulfide form; in the sulfur-free state the catalytic Cys residue adopts two alternate conformations, reflected by perturbation of the neighboring active-site residues, which is associated with a partly reversible loss of thiosulfate:cyanide sulfurtransferase activity. The catalytic mechanism of A. vinelandii rhodanese relies primarily on the main-chain conformation of the 230 to 235 active-site loop and on a surrounding strong positive electrostatic field. Substrate recognition is based on residues which are entirely different in the prokaryotic and eukaryotic enzymes. The active-site loop of A. vinelandii rhodanese displays striking structural similarity to the active-site loop of the similarly folded catalytic domain of dual specific phosphatase Cdc25, suggesting a common evolutionary origin of the two enzyme families. PMID- 10788331 TI - Pathways of ligand clearance in acetylcholinesterase by multiple copy sampling. AB - The clearance of seven different ligands from the deeply buried active-site of Torpedo californica acetylcholinesterase is investigated by combining multiple copy sampling molecular dynamics simulations, with the analysis of protein-ligand interactions, protein motion and the electrostatic potential sampled by the ligand copies along their journey outwards. The considered ligands are the cations ammonium, methylammonium, and tetramethylammonium, the hydrophobic methane and neopentane, and the anionic product acetate and its neutral form, acetic acid. We find that the pathways explored by the different ligands vary with ligand size and chemical properties. Very small ligands, such as ammonium and methane, exit through several routes. One involves the main exit through the mouth of the enzyme gorge, another is through the so-called back door near Trp84, and a third uses a side door at a direction of approximately 45 degrees to the main exit. The larger polar ligands, methylammonium and acetic acid, leave through the main exit, but the bulkiest, tetramethylammonium and neopentane, as well as the smaller acetate ion, remain trapped in the enzyme gorge during the time of the simulations. The pattern of protein-ligand contacts during the diffusion process is highly non-random and differs for different ligands. A majority is made with aromatic side-chains, but classical H-bonds are also formed. In the case of acetate, but not acetic acid, the anionic and neutral form, respectively, of one of the reaction products, specific electrostatic interactions with protein groups, seem to slow ligand motion and interfere with protein flexibility; protonation of the acetate ion is therefore suggested to facilitate clearance. The Poisson-Boltzmann formalism is used to compute the electrostatic potential of the thermally fluctuating acetylcholinesterase protein at positions actually visited by the diffusing ligand copies. Ligands of different charge and size are shown to sample somewhat different electrostatic potentials during their migration, because they explore different microscopic routes. The potential along the clearance route of a cation such as methylammonium displays two clear minima at the active and peripheral anionic site. We find moreover that the electrostatic energy barrier that the cation needs to overcome when moving between these two sites is small in both directions, being of the order of the ligand kinetic energy. The peripheral site thus appears to play a role in trapping inbound cationic ligands as well as in cation clearance, and hence in product release. PMID- 10788332 TI - Exploiting excess sharing: a more powerful test of linkage for affected sib pairs than the transmission/disequilibrium test. AB - The transmission/disequilibrium test (TDT) is a popular, simple, and powerful test of linkage, which can be used to analyze data consisting of transmissions to the affected members of families with any kind pedigree structure, including affected sib pairs (ASPs). Although it is based on the preferential transmission of a particular marker allele across families, it is not a valid test of association for ASPs. Martin et al. devised a similar statistic for ASPs, Tsp, which is also based on preferential transmission of a marker allele but which is a valid test of both linkage and association for ASPs. It is, however, less powerful than the TDT as a test of linkage for ASPs. What I show is that the differences between the TDT and Tsp are due to the fact that, although both statistics are based on preferential transmission of a marker allele, the TDT also exploits excess sharing in identity-by-descent transmissions to ASPs. Furthermore, I show that both of these statistics are members of a family of "TDT like" statistics for ASPs. The statistics in this family are based on preferential transmission but also, to varying extents, exploit excess sharing. From this family of statistics, we see that, although the TDT exploits excess sharing to some extent, it is possible to do so to a greater extent-and thus produce a more powerful test of linkage, for ASPs, than is provided by the TDT. Power simulations conducted under a number of disease models are used to verify that the most powerful member of this family of TDT-like statistics is more powerful than the TDT for ASPs. PMID- 10788333 TI - Candidate locus for a nuclear modifier gene for maternally inherited deafness. AB - Maternally inherited deafness associated with the A1555G mutation in the mitochondrial 12S ribosomal RNA (rRNA) gene appears to require additional environmental or genetic changes for phenotypic expression. Aminoglycosides have been identified as one such environmental factor. In one large Arab-Israeli pedigree with congenital hearing loss in some of the family members with the A1555G mutation and with no exposure to aminoglycosides, biochemical evidence has suggested the role of nuclear modifier gene(s), but a genomewide search has indicated the absence of a single major locus having such an effect. Thus it has been concluded that the penetrance of the mitochondrial mutation appears to depend on additive effects of several nuclear genes. We have now investigated 10 multiplex Spanish and Italian families with 35 members with the A1555G mutation and sensorineural deafness. Parametric analysis of a genomewide screen again failed to identify significant evidence for linkage to a single autosomal locus. However, nonparametric analysis supported the role of the chromosomal region around marker D8S277. The combined maximized allele-sharing LOD score of 3.1 in Arab-Israeli/Spanish/Italian families represents a highly suggestive linkage result. We suggest that this region should be considered a candidate for containing the first human nuclear modifier gene for a mitochondrial DNA disorder. The locus operates in Arab-Israeli, Spanish, and Italian families, resulting in the deafness phenotype on a background of the mitochondrial A1555G mutation. No obvious candidate genes are located in this region. PMID- 10788334 TI - Founder mutations in the BRCA1 gene in Polish families with breast-ovarian cancer. AB - We have undertaken a hospital-based study, to identify possible BRCA1 and BRCA2 founder mutations in the Polish population. The study group consisted of 66 Polish families with cancer who have at least three related females affected with breast or ovarian cancer and who had cancer diagnosed, in at least one of the three affected females, at age <50 years. A total of 26 families had both breast and ovarian cancers, 4 families had ovarian cancers only, and 36 families had breast cancers only. Genomic DNA was prepared from the peripheral blood leukocytes of at least one affected woman from each family. The entire coding region of BRCA1 and BRCA2 was screened for the presence of germline mutations, by use of SSCP followed by direct sequencing of observed variants. Mutations were found in 35 (53%) of the 66 families studied. All but one of the mutations were detected within the BRCA1 gene. BRCA1 abnormalities were identified in all four families with ovarian cancer only, in 67% of 27 families with both breast and ovarian cancer, and in 34% of 35 families with breast cancer only. The single family with a BRCA2 mutation had the breast-ovarian cancer syndrome. Seven distinct mutations were identified; five of these occurred in two or more families. In total, recurrent mutations were found in 33 (94%) of the 35 families with detected mutations. Three BRCA1 abnormalities-5382insC, C61G, and 4153delA accounted for 51%, 20%, and 11% of the identified mutations, respectively. PMID- 10788335 TI - A novel disorder caused by defective biosynthesis of N-linked oligosaccharides due to glucosidase I deficiency. AB - Glucosidase I is an important enzyme in N-linked glycoprotein processing, removing specifically distal alpha-1,2-linked glucose from the Glc3Man9GlcNAc2 precursor after its en bloc transfer from dolichyl diphosphate to a nascent polypeptide chain in the endoplasmic reticulum. We have identified a glucosidase I defect in a neonate with severe generalized hypotonia and dysmorphic features. The clinical course was progressive and was characterized by the occurrence of hepatomegaly, hypoventilation, feeding problems, seizures, and fatal outcome at age 74 d. The accumulation of the tetrasaccharide Glc(alpha1-2)Glc(alpha1 3)Glc(alpha1-3)Man in the patient's urine indicated a glycosylation disorder. Enzymological studies on liver tissue and cultured skin fibroblasts revealed a severe glucosidase I deficiency. The residual activity was <3% of that of controls. Glucosidase I activities in cultured skin fibroblasts from both parents were found to be 50% of those of controls. Tissues from the patient subjected to SDS-PAGE followed by immunoblotting revealed strongly decreased amounts of glucosidase I protein in the homogenate of the liver, and a less-severe decrease in cultured skin fibroblasts. Molecular studies showed that the patient was a compound heterozygote for two missense mutations in the glucosidase I gene: (1) one allele harbored a G-->C transition at nucleotide (nt) 1587, resulting in the substitution of Arg at position 486 by Thr (R486T), and (2) on the other allele a T-->C transition at nt 2085 resulted in the substitution of Phe at position 652 by Leu (F652L). The mother was heterozygous for the G-->C transition, whereas the father was heterozygous for the T-->C transition. These base changes were not seen in 100 control DNA samples. A causal relationship between the alpha glucosidase I deficiency and the disease is postulated. PMID- 10788336 TI - Genomic differentiation of Neanderthals and anatomically modern man allows a fossil-DNA-based classification of morphologically indistinguishable hominid bones. AB - Southern blot hybridizations of genomic DNA were introduced as a relatively simple fossil-DNA-based approach to classify remains of Neanderthals. When hybridized with genomic DNA of either human or Neanderthal origin, DNA extracted from two Neanderthal finds-the Os parietale, from Warendorf-Neuwarendorf, Germany, and a clavicula, from Krapina, Croatia-was shown to yield hybridization signals that differ by at least a factor of two compared to the signals obtained with the use of fossil DNA of an early Homo sapiens from the Vogelherd cave (Stetten I), Germany. When labeled chimpanzee DNA was used as a probe, Neanderthal and human DNA, however, revealed hybridization signals of similar intensity. Thus, the genome of Neanderthals is expected to differ significantly from the genome of anatomically modern man, because of the contrasting composition of repetitive DNA. These data support the hypothesis that Neanderthals were not ancestors of anatomically modern man. PMID- 10788338 TI - Immobilization with metal hydroxides as a means to concentrate food-borne bacteria for detection by cultural and molecular methods. AB - The application of nucleic acid amplification methods to the detection of food borne pathogens could be facilitated by concentrating the organisms from the food matrix before detection. This study evaluated the utility of metal hydroxide immobilization for the concentration of bacterial cells from dairy foods prior to detection by cultural and molecular methods. Using reconstituted nonfat dry milk (NFDM) as a model, two food-borne pathogens (Listeria monocytogenes and Salmonella enterica serovar Enteritidis) were concentrated from 25-ml samples by the sequential steps of clarification and high-speed centrifugation (designated primary concentration) and immobilization with zirconium hydroxide and low-speed centrifugation (designated secondary concentration). Sample volume reduction after immobilization with zirconium hydroxide was 50-fold, with total bacterial recoveries ranging from 78 to 96% of input for serovar Enteritidis and 65 to 96% of input for L. monocytogenes. Immobilized bacteria remained viable and could be enumerated by standard cultural procedures. When followed by RNA extraction and subsequent detection by reverse transcription (RT)-PCR, detection limits of 10(1) to 10(2) CFU/25 ml of reconstituted NFDM were achieved for both organisms. The bacterial-immobilization step was relatively nonspecific, resulting in recovery of >50% of the input cells when evaluated on a panel of representative bacterial strains of significance to foods. The method could be adapted to more complex dairy products, such as whole milk and ice cream, for which bacterial recoveries after immobilization ranged from 64 to >100%, with subsequent RT-PCR detection limits of >/=10(2) CFU/ml for whole milk and >/=10(1) CFU for ice cream for both serovar Enteritidis and L. monocytogenes. The bacterial-immobilization method is easy, rapid, and inexpensive and may have applications for the concentration of a wide variety of food-borne bacteria prior to detection by both conventional and alternative methods. PMID- 10788337 TI - Haplotypes and linkage disequilibrium at the phenylalanine hydroxylase locus, PAH, in a global representation of populations. AB - Because defects in the phenylalanine hydroxylase gene (PAH) cause phenylketonuria (PKU), PAH was studied for normal polymorphisms and linkage disequilibrium soon after the gene was cloned. Studies in the 1980s concentrated on European populations in which PKU was common and showed that haplotype-frequency variation exists between some regions of the world. In European populations, linkage disequilibrium generally was found not to exist between RFLPs at opposite ends of the gene but was found to exist among the RFLPs clustered at each end. We have now undertaken the first global survey of normal variation and disequilibrium across the PAH gene. Four well-mapped single-nucleotide polymorphisms (SNPs) spanning approximately 75 kb, two near each end of the gene, were selected to allow linkage disequilibrium across most of the gene to be examined. These SNPs were studied as PCR-RFLP markers in samples of, on average, 50 individuals for each of 29 populations, including, for the first time, multiple populations from Africa and from the Americas. All four sites are polymorphic in all 29 populations. Although all but 5 of the 16 possible haplotypes reach frequencies >5% somewhere in the world, no haplotype was seen in all populations. Overall linkage disequilibrium is highly significant in all populations, but disequilibrium between the opposite ends is significant only in Native American populations and in one African population. This study demonstrates that the physical extent of linkage disequilibrium can differ substantially among populations from different regions of the world, because of both ancient genetic drift in the ancestor common to a large regional group of modern populations and recent genetic drift affecting individual populations. PMID- 10788339 TI - A few cosmopolitan phylotypes dominate planktonic archaeal assemblages in widely different oceanic provinces. AB - We compared the phylogenetic compositions of marine planktonic archaeal populations in different marine provinces. Samples from eight different environments were collected at two depths (surface and aphotic zone), and 16 genetic libraries of PCR-amplified archaeal 16S rRNA genes were constructed. The libraries were analyzed by using a three-step hierarchical approach. Membrane hybridization experiments revealed that most of the archaeal clones were affiliated with one of the two groups of marine archaea described previously, crenarchaeotal group I and euryarchaeotal group II. One of the 2,328 ribosomal DNA clones analyzed was related to a different euryarchaeal lineage, which was recently recovered from deep-water marine plankton. In temperate regions (Pacific Ocean, Atlantic Ocean, and Mediterranean Sea) both major groups were found at the two depths investigated; group II predominated at the surface, and group I predominated at depth. In Antarctic and subantarctic waters group II was practically absent. The clonal compositions of archaeal libraries were investigated by performing a restriction fragment length polymorphism (RFLP) analysis with two tetrameric restriction enzymes, which defined discrete operational taxonomic units (OTUs). The OTUs defined in this way were phylogenetically consistent; clones belonging to the same OTU were closely related. The clonal diversity as determined by the RFLP analysis was low, and most libraries were dominated by only one or two OTUs. Some OTUs were found in samples obtained from very distant places, indicating that some phylotypes were ubiquitous. A tree containing one example of each OTU detected was constructed, and this tree revealed that there were several clusters within archaeal group I and group II. The members of some of these clusters had different depth distributions. PMID- 10788340 TI - Purification to homogeneity and characterization of a novel Pseudomonas putida chromate reductase. AB - Cr(VI) (chromate) is a widespread environmental contaminant. Bacterial chromate reductases can convert soluble and toxic chromate to the insoluble and less toxic Cr(III). Bioremediation can therefore be effective in removing chromate from the environment, especially if the bacterial propensity for such removal is enhanced by genetic and biochemical engineering. To clone the chromate reductase-encoding gene, we purified to homogeneity (>600-fold purification) and characterized a novel soluble chromate reductase from Pseudomonas putida, using ammonium sulfate precipitation (55 to 70%), anion-exchange chromatography (DEAE Sepharose CL-6B), chromatofocusing (Polybuffer exchanger 94), and gel filtration (Superose 12 HR 10/30). The enzyme activity was dependent on NADH or NADPH; the temperature and pH optima for chromate reduction were 80 degrees C and 5, respectively; and the K(m) was 374 microM, with a V(max) of 1.72 micromol/min/mg of protein. Sulfate inhibited the enzyme activity noncompetitively. The reductase activity remained virtually unaltered after 30 min of exposure to 50 degrees C; even exposure to higher temperatures did not immediately inactivate the enzyme. X-ray absorption near-edge-structure spectra showed quantitative conversion of chromate to Cr(III) during the enzyme reaction. PMID- 10788341 TI - In situ reverse transcription-PCR for monitoring gene expression in individual Methanosarcina mazei S-6 cells. AB - An in situ reverse transcription-PCR protocol for detecting specific mRNA in Methanosarcina mazei S-6 is described. This method allowed us to detect heat shock-induced increases in the intracellular levels of the transcript of the universal stress gene dnaK. The cell walls of paraformaldehyde-fixed cells were permeabilized by a thermal cycling procedure or by lysozyme treatment, and the cellular DNA was removed with DNase. The cells were subjected to a seminested reverse transcription-PCR protocol in which a digoxigenin-labeled primer was used. Detection of the reporter molecule was based on the 2-hydroxy-3-naphtoic acid-2'-phenylanilide phosphate-Fast Red detection system and binding of anti digoxigenin-alkaline phosphatase conjugate. Fluorescence in permeabilized cells increased after a heat shock compared to fluorescence in non-heat-shocked cells, and the increase corresponded to an increase in the level of the dnaK transcript. PMID- 10788342 TI - Molecular analyses of novel methanotrophic communities in forest soil that oxidize atmospheric methane. AB - Forest and other upland soils are important sinks for atmospheric CH(4), consuming 20 to 60 Tg of CH(4) per year. Consumption of atmospheric CH(4) by soil is a microbiological process. However, little is known about the methanotrophic bacterial community in forest soils. We measured vertical profiles of atmospheric CH(4) oxidation rates in a German forest soil and characterized the methanotrophic populations by PCR and denaturing gradient gel electrophoresis (DGGE) with primer sets targeting the pmoA gene, coding for the alpha subunit of the particulate methane monooxygenase, and the small-subunit rRNA gene (SSU rDNA) of all life. The forest soil was a sink for atmospheric CH(4) in situ and in vitro at all times. In winter, atmospheric CH(4) was oxidized in a well-defined subsurface soil layer (6 to 14 cm deep), whereas in summer, the complete soil core was active (0 cm to 26 cm deep). The content of total extractable DNA was about 10-fold higher in summer than in winter. It decreased with soil depth (0 to 28 cm deep) from about 40 to 1 microg DNA per g (dry weight) of soil. The PCR product concentration of SSU rDNA of all life was constant both in winter and in summer. However, the PCR product concentration of pmoA changed with depth and season. pmoA was detected only in soil layers with active CH(4) oxidation, i.e., 6 to 16 cm deep in winter and throughout the soil core in summer. The same methanotrophic populations were present in winter and summer. Layers with high CH(4) consumption rates also exhibited more bands of pmoA in DGGE, indicating that high CH(4) oxidation activity was positively correlated with the number of methanotrophic populations present. The pmoA sequences derived from excised DGGE bands were only distantly related to those of known methanotrophs, indicating the existence of unknown methanotrophs involved in atmospheric CH(4) consumption. PMID- 10788343 TI - (1-->6)-beta-D-glucan as cell wall receptor for Pichia membranifaciens killer toxin. AB - The killer toxin from Pichia membranifaciens CYC 1106, a yeast isolated from fermenting olive brines, binds primarily to the (1-->6)-beta-D-glucan of the cell wall of a sensitive yeast (Candida boidinii IGC 3430). The (1-->6)-beta-D-glucan was purified from cell walls of C. boidinii by alkali and hot-acetic acid extraction, a procedure which solubilizes glucans. The major fraction of receptor activity remained with the alkali-insoluble (1-->6)-beta- and (1-->3)-beta-D glucans. The chemical (gas-liquid chromatography) and structural (periodate oxidation, infrared spectroscopy, and (1)H nuclear magnetic resonance) analyses of the fractions obtained showed that (1-->6)-beta-D-glucan was a receptor. Adsorption of most of the killer toxin to the (1-->6)-beta-D-glucan was complete within 2 min. Killer toxin adsorption to the linear (1-->6)-beta-D-glucan, pustulan, and a glucan from Penicillium allahabadense was observed. Other polysaccharides with different linkages failed to bind the killer toxin. The specificity of the killer toxin for its primary receptor provides an effective means to purify the killer toxin, which may have industrial applications for fermentations in which salt is present as an adjunct, such as olive brines. This toxin shows its maximum killer activity in the presence of NaCl. This report is the first to identify the (1-->6)-beta-D-glucan as a receptor for this novel toxin. PMID- 10788344 TI - Quantification of phnAc and nahAc in contaminated new zealand soils by competitive PCR. AB - Unculturable polycyclic aromatic hydrocarbon (PAH)-degrading bacteria are a significant reservoir of the microbial potential to catabolize low-molecular weight PAHs. The population of these bacteria is larger than the population of nah-like bacteria that are the dominant organisms in culture-based studies. We used the recently described phn genes of Burkholderia sp. strain RP007, which feature only rarely in culture-based studies, as an alternative genotype for naphthalene and phenanthrene degradation and compared this genotype with the genotypically distinct but ubiquitous nah-like class in different soils. Competitive PCR quantification of phnAc and nahAc, which encode the iron sulfur protein large (alpha) subunits of PAH dioxygenases in nah-like and phn catabolic operons, revealed that the phn genotype can have a greater ecological significance than the nah-like genotype. PMID- 10788345 TI - Characterization of plasmid-borne and chromosome-encoded traits of Agrobacterium biovar 1, 2, and 3 strains from France. AB - We collected 111 Agrobacterium isolates from galls of various origins (most of them from France) and analyzed both their plasmid-borne and chromosome-encoded traits. Phenotypic analysis of these strains allowed their classification in three phena which exactly matched the delineation of biovars 1, 2, and 3. A fourth phenon was identified which comprises three atypical strains. The phenotypic analysis has also allowed us to identify 12 additional characteristics which could be used to identify the three biovars of Agrobacterium. Our results also suggest that biovar 1 and 2 represent distinct species. Analysis of plasmid borne traits confirmed that tartrate utilization is a common feature of biovar 3 strains (now named Agrobacterium vitis) and of Agrobacterium grapevine strains in general. Among pathogenic strains of Agrobacterium, several exhibited unusual opine synthesis and degradation patterns, and one strain of biovar 3 induced tumors containing vitopine and a novel opine-like molecule derived from putrescine. We have named this compound rideopine. PMID- 10788346 TI - The chromosomal arsenic resistance genes of Thiobacillus ferrooxidans have an unusual arrangement and confer increased arsenic and antimony resistance to Escherichia coli. AB - The chromosomal arsenic resistance genes of the acidophilic, chemolithoautotrophic, biomining bacterium Thiobacillus ferrooxidans were cloned and sequenced. Homologues of four arsenic resistance genes, arsB, arsC, arsH, and a putative arsR gene, were identified. The T. ferrooxidans arsB (arsenite export) and arsC (arsenate reductase) gene products were functional when they were cloned in an Escherichia coli ars deletion mutant and conferred increased resistance to arsenite, arsenate, and antimony. Therefore, despite the fact that the ars genes originated from an obligately acidophilic bacterium, they were functional in E. coli. Although T. ferrooxidans is gram negative, its ArsC was more closely related to the ArsC molecules of gram-positive bacteria. Furthermore, a functional trxA (thioredoxin) gene was required for ArsC-mediated arsenate resistance in E. coli; this finding confirmed the gram-positive ArsC-like status of this resistance and indicated that the division of ArsC molecules based on Gram staining results is artificial. Although arsH was expressed in an E. coli derived in vitro transcription-translation system, ArsH was not required for and did not enhance arsenic resistance in E. coli. The T. ferrooxidans ars genes were arranged in an unusual manner, and the putative arsR and arsC genes and the arsBH genes were translated in opposite directions. This divergent orientation was conserved in the four T. ferrooxidans strains investigated. PMID- 10788347 TI - Isolation of adherent polycyclic aromatic hydrocarbon (PAH)-degrading bacteria using PAH-sorbing carriers. AB - Two different procedures were compared to isolate polycyclic aromatic hydrocarbon (PAH)-utilizing bacteria from PAH-contaminated soil and sludge samples, i.e., (i) shaken enrichment cultures in liquid mineral medium in which PAHs were supplied as crystals and (ii) a new method in which PAH degraders were enriched on and recovered from hydrophobic membranes containing sorbed PAHs. Both techniques were successful, but selected from the same source different bacterial strains able to grow on PAHs as the sole source of carbon and energy. The liquid enrichment mainly selected for Sphingomonas spp., whereas the membrane method exclusively led to the selection of Mycobacterium spp. Furthermore, in separate membrane enrichment set-ups with different membrane types, three repetitive extragenic palindromic PCR-related Mycobacterium strains were recovered. The new Mycobacterium isolates were strongly hydrophobic and displayed the capacity to adhere strongly to different surfaces. One strain, Mycobacterium sp. LB501T, displayed an unusual combination of high adhesion efficiency and an extremely high negative charge. This strain may represent a new bacterial species as suggested by 16S rRNA gene sequence analysis. These results indicate that the provision of hydrophobic sorbents containing sorbed PAHs in the enrichment procedure discriminated in favor of certain bacterial characteristics. The new isolation method is appropriate to select for adherent PAH-degrading bacteria, which might be useful to biodegrade sorbed PAHs in soils and sludge. PMID- 10788348 TI - Metabolic analysis of Escherichia coli in the presence and absence of the carboxylating enzymes phosphoenolpyruvate carboxylase and pyruvate carboxylase. AB - Fermentation patterns of Escherichia coli with and without the phosphoenolpyruvate carboxylase (PPC) and pyruvate carboxylase (PYC) enzymes were compared under anaerobic conditions with glucose as a carbon source. Time profiles of glucose and fermentation product concentrations were determined and used to calculate metabolic fluxes through central carbon pathways during exponential cell growth. The presence of the Rhizobium etli pyc gene in E. coli (JCL1242/pTrc99A-pyc) restored the succinate producing ability of E. coli ppc null mutants (JCL1242), with PYC competing favorably with both pyruvate formate lyase and lactate dehydrogenase. Succinate formation was slightly greater by JCL1242/pTrc99A-pyc than by cells which overproduced PPC (JCL1242/pPC201, ppc(+)), even though PPC activity in cell extracts of JCL1242/pPC201 (ppc(+)) was 40-fold greater than PYC activity in extracts of JCL1242/pTrc99a-pyc. Flux calculations indicate that during anaerobic metabolism the pyc(+) strain had a 34% greater specific glucose consumption rate, a 37% greater specific rate of ATP formation, and a 6% greater specific growth rate compared to the ppc(+) strain. In light of the important position of pyruvate at the juncture of NADH-generating pathways and NADH-dissimilating branches, the results show that when PPC or PYC is expressed, the metabolic network adapts by altering the flux to lactate and the molar ratio of ethanol to acetate formation. PMID- 10788349 TI - Use of an enzyme-linked lectinsorbent assay to monitor the shift in polysaccharide composition in bacterial biofilms. AB - An enzyme-linked lectinsorbent assay (ELLA) was developed for quantification and characterization of extracellular polysaccharides produced by 1- and 4-day biofilms of 10 bacterial strains isolated from food industry premises. Peroxidase labeled concanavalin A (ConA) and wheat germ agglutinin (WGA) were used, as they specifically bind to saccharide residues most frequently encountered in biofilms matrices: D-glucose or D-mannose for ConA and N-acetyl-D-glucosamine or N acetylneuraminic acid for WGA. The ELLA applied to 1- and 4-day biofilms colonizing wells of microtiter plates was able to detect that for Stenotrophomonas maltophilia and to a lesser extent Staphylococcus sciuri, the increase in production of exopolysaccharides over time was not the same for sugars binding with ConA and those binding with WGA. Differences in extracellular polysaccharides produced were observed among strains belonging to the same species. These results demonstrate that ELLA is a useful tool not only for rapid characterization of biofilm extracellular polysaccharides but also, in studies of individual strains, for detection of changes over time in the proportion of the exopolysaccharidic component within the polymeric matrix. PMID- 10788350 TI - Viral density and virus-to-bacterium ratio in deep-sea sediments of the Eastern Mediterranean. AB - Viruses are now recognized as a key component in pelagic systems, but their role in marine sediment has yet to be assessed. In this study bacterial and viral densities were determined at nine deep-sea stations selected from three main sites (i.e., the Sporades Basin, the Cretan Sea, and the Ierapetra Trench at depths of 1,232, 1,840, and 4,235 m, respectively) of the Eastern Mediterranean. The three areas were characterized by different phytopigment and biopolymeric carbon concentrations and by changes in the protein and carbohydrate pools. A gradient of increasing trophic conditions was observed from the Sporades Basin (North Aegean) to the Ierapetra Trench (South Aegean). Viral densities (ranging from 1 x 10(9) to 2 x 10(9) viruses ml of sediment(-1)) were significantly correlated to bacterial densities (n = 9, r(2) = 0.647) and reached values up to 3 orders of magnitude higher than those generally reported for the water column. However, the virus-to-bacterium density ratio in deep-sea sediments was about 1 order of magnitude lower (range of 2 to 5, with a modal value of 2.6) than in pelagic environments. Virus density decreased vertically with depth in sediment cores at all stations and was below detection limits at the 10-cm depth of the abyssal sediments of the Ierapetra Trench. Virus density in the sediment apparently reflected a gradient of particle fluxes and trophic conditions, displaying the highest values in the Sporades Basin. The low virus-to-bacterium ratios and their inverse relationship with station depth suggest that the role played by viruses in controlling deep-sea benthic bacterial assemblages and biogeochemical cycles is less relevant than in pelagic systems. PMID- 10788351 TI - Increased killing of Bacillus subtilis on the hair roots of transgenic T4 lysozyme-producing potatoes. AB - Transgenic potato plants expressing the phage T4 lysozyme gene which are resistant to the plant-pathogenic enterobacterium Erwinia carotovora subsp. carotovora have been constructed. The agricultural growth of these potatoes might have harmful effects on soil microbiota as a result of T4 lysozyme release into the rhizosphere. To assess the bactericidal effect of roots, we have developed a novel method to associate the cells of Bacillus subtilis with hair roots of plants and to quantify the survival of cells directly on the root surface by appropriate staining and fluorescence microscopy. With this technique, we found that the roots of potato plants (Desiree and transgenic control lines) without T4 lysozyme gene display measurable killing activity on root-adsorbed B. subtilis cells. Killing was largely independent of the plant age and growth of plants in greenhouse or field plots. Roots from potato lines expressing the T4 lysozyme gene always showed significantly (1.5- to 3.5-fold) higher killing. It is concluded that T4 lysozyme is released from the root epidermis cells and is active in the fluid film on the root surface. We discuss why strong negative effects of T4 lysozyme-producing potatoes on soil bacteria in field trials may not be observed. We propose that the novel method presented here to study interactions of bacteria with roots can be applied not only to bacterial killing but also to interactions leading to growth-sustaining effects of plants on bacteria. PMID- 10788352 TI - Detection of infectious Cryptosporidium parvum oocysts in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule). AB - Infective Cryptosporidium parvum oocysts were detected in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) from a shellfish-producing region (Gallaecia, northwest Spain, bounded by the Atlantic Ocean) that accounts for the majority of European shellfish production. Shellfish were collected from bay sites with different degrees of organic pollution. Shellfish harboring C. parvum oocysts were recovered only from areas located near the mouths of rivers with a high density of grazing ruminants on their banks. An approximation of the parasite load of shellfish collected in positive sites indicated that each shellfish transported more than 10(3) oocysts. Recovered oocysts were infectious for neonatal mice, and PCR-restriction fragment length polymorphism analysis demonstrated a profile similar to that described for genotype C or 2 of the parasite. These results demonstrate that mussels and cockles could act as a reservoir of C. parvum infection for humans. Moreover, estuarine shellfish could be used as an indicator of river water contamination. PMID- 10788353 TI - Carbon monoxide dehydrogenase activity in Bradyrhizobium japonicum. AB - Bradyrhizobium japonicum strain 110spc4 was capable of chemolithoautotrophic growth with carbon monoxide (CO) as a sole energy and carbon source under aerobic conditions. The enzyme carbon monoxide dehydrogenase (CODH; EC 1.2.99.2) has been purified 21-fold, with a yield of 16% and a specific activity of 58 nmol of CO oxidized/min/mg of protein, by a procedure that involved differential ultracentrifugation, anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. The purified enzyme gave a single protein and activity band on nondenaturing polyacrylamide gel electrophoresis and had a molecular mass of 230,000 Da. The 230-kDa enzyme was composed of large (L; 75 kDa), medium (M; 28.4-kDa), and small (S; 17.2-kDa) subunits occurring in heterohexameric (LMS)(2) subunit composition. The 75-kDa polypeptide exhibited immunological cross-reactivity with the large subunit of the CODH of Oligotropha carboxidovorans. The B. japonicum enzyme contained, per mole, 2.29 atoms of Mo, 7.96 atoms of Fe, 7.60 atoms of labile S, and 1.99 mol of flavin. Treatment of the enzyme with iodoacetamide yielded di(carboxamidomethyl)molybdopterin cytosine dinucleotide, identifying molybdopterin cytosine dinucleotide as the organic portion of the B. japonicum CODH molybdenum cofactor. The absorption spectrum of the purified enzyme was characteristic of a molybdenum-containing iron-sulfur flavoprotein. PMID- 10788354 TI - Toluene monooxygenase-catalyzed epoxidation of alkenes. AB - Several toluene monooxygenase-producing organisms were tested for their ability to oxidize linear alkenes and chloroalkenes three to eight carbons long. Each of the wild-type organisms degraded all of the alkenes that were tested. Epoxides were produced during the oxidation of butene, butadiene, and pentene but not hexene or octadiene. A strain of Escherichia coli expressing the cloned toluene-4 monooxygenase (T4MO) of Pseudomonas mendocina KR1 was able to oxidize butene, butadiene, pentene, and hexene but not octadiene, producing epoxides from all of the substrates that were oxidized. A T4MO-deficient variant of P. mendocina KR1 oxidized alkenes that were five to eight carbons long, but no epoxides were detected, suggesting the presence of multiple alkene-degrading enzymes in this organism. The alkene oxidation rates varied widely (ranging from 0. 01 to 0.33 micromol of substrate/min/mg of cell protein) and were specific for each organism substrate pair. The enantiomeric purity of the epoxide products also varied widely, ranging from 54 to >90% of a single epoxide enantiomer. In the absence of more preferred substrates, such as toluene or alkenes, the epoxides underwent further toluene monooxygenase-catalyzed transformations, forming products that were not identified. PMID- 10788355 TI - Cloning and sequencing of the gene encoding an aldehyde dehydrogenase that is induced by growing Alteromonas sp. Strain KE10 in a low concentration of organic nutrients. AB - The protein composition of Alteromonas sp. strain KE10 cultured at two different organic-nutrient concentrations was determined by using two-dimensional polyacrylamide gel electrophoresis. The cellular levels of three proteins, OlgA, B, and -C, were considerably higher in cells grown in a low concentration of organic nutrient medium (LON medium; 0.2 mg of carbon per liter) than cells grown in a high concentration of organic nutrient medium (HON; 200 mg of C liter(-1)) or cells starved for organic nutrients. In the LON medium, the cellular levels of the Olg proteins were higher at the exponential growth phase than at the stationary growth phase. A sequence of the gene for OlgA revealed that the amino acid sequence had a high degree of similarity to the NAD(+)-dependent aldehyde dehydrogenases of several bacteria. OlgA, expressed in Escherichia coli, catalyzed the dehydrogenation of acetaldehyde, propionaldehyde, and butyraldehyde. The aldehyde dehydrogenase activity of KE10 was higher in cells growing exponentially in LON medium than in HON. OlgA may be involved in the growth under low-nutrient conditions. The physiological role of OlgA is discussed here. PMID- 10788356 TI - Molecular characterization and identification of biocontrol isolates of Trichoderma spp. AB - The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as "Trichoderma harzianum" Rifai and one biocontrol strain recognized as T. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains in T. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae, Rosellinia necatrix, Botrytis cinerea, and Fusarium oxysporum f. sp. dianthi in three different media. PMID- 10788357 TI - Mycotoxins in crude building materials from water-damaged buildings. AB - We analyzed 79 bulk samples of moldy interior finishes from Finnish buildings with moisture problems for 17 mycotoxins, as well as for fungi that could be isolated using one medium and one set of growth conditions. We found the aflatoxin precursor, sterigmatocystin, in 24% of the samples and trichothecenes in 19% of the samples. Trichothecenes found included satratoxin G or H in five samples; diacetoxyscirpenol in five samples; and 3-acetyl-deoxynivalenol, deoxynivalenol, verrucarol, or T-2-tetraol in an additional five samples. Citrinine was found in three samples. Aspergillus versicolor was present in most sterigmatocystin-containing samples, and Stachybotrys spp. were present in the samples where satratoxins were found. In many cases, however, the presence of fungi thought to produce the mycotoxins was not correlated with the presence of the expected compounds. However, when mycotoxins were found, some toxigenic fungi usually were present, even if the species originally responsible for producing the mycotoxin was not isolated. We conclude that the identification and enumeration of fungal species present in bulk materials are important to verify the severity of mold damage but that chemical analyses are necessary if the goal is to establish the presence of mycotoxins in moldy materials. PMID- 10788358 TI - Enantiomeric degradation of 2-(4-Sulfophenyl)Butyrate via 4-sulfocatechol in Delftia acidovorans SPB1. AB - Enrichment cultures with enantiomeric 2-(4-sulfophenyl)butyrate (SPB) as the sole added source(s) of carbon and energy for growth yielded a pure culture of a degradative bacterium, which was identified as Delftia acidovorans SPB1. The organism utilized the enantiomers sequentially. R-SPB was utilized first (specific growth rate [mu] = 0.28 h(-1)), with transient excretion of an unknown intermediate, which was identified as 4-sulfocatechol (4SC). Utilization of S-SPB was slower (mu = 0.016 h(-1)) and was initiated only after the first enantiomer was exhausted. Suspensions of cells grown in S-SPB excreted 4SC, so metabolism of the two enantiomers converged at 4SC. The latter was degraded by ortho cleavage via 3-sulfo-cis,cis-muconate. Strain SPB1 grew with 4SC and with 1-(4 sulfophenyl)octane (referred to herein as model LAS) but not with commercial linear alkylbenzenesulfonate (LAS) surfactant, which is subterminally substituted but nontoxic. It would appear that metabolism of the model LAS does not represent metabolism of commercial LAS. PMID- 10788359 TI - An alpha-proteobacterium converts linear alkylbenzenesulfonate surfactants into sulfophenylcarboxylates and linear alkyldiphenyletherdisulfonate surfactants into sulfodiphenylethercarboxylates. AB - The surfactant linear alkylbenzenesulfonate (LAS; 0.5 mM) or linear monoalkyldiphenyletherdisulfonate (LADPEDS; 0.5 mM) in salts medium was easily degraded in laboratory trickling filters, whereas carbon-limited, aerobic enrichment cultures in suspended culture with the same inocula did not grow. We took portions of the trickling filters which degraded LADPEDS, shook the organisms from the solid support (polyester), and found that growth in suspended culture in LADPEDS-salts medium occurred only in the presence of some solid support (polyester fleece or glass wool), though little biomass was immobilized on the support. The end products in suspended culture were identical with those from the trickling filters. There was low plating efficiency of LADPEDS-grown cultures on complex medium, and no picked colony or mixture of colonies grew in LADPEDS-salts-glass wool medium. However, selective plates containing LADPEDS salts medium solidified with agarose yielded LADPEDS-dependent, pinpoint colonies which could be picked singly and subcultured in selective liquid medium. Isolate DS-1 was a bacterium which showed 93% sequence homology (16S ribosomal DNA) to its nearest phylogenetic neighbor, an alpha-proteobacterium. Strain DS-1 grew heterotrophically in LADPEDS-salts-glass wool medium and converted the set of aryl-substituted alkanes to the corresponding aryl-substituted carboxylic acids of shorter chain length. Similarly, strain DS-1 grew heterotrophically with commercial LAS, converting it to a set of sulfophenylcarboxylates. Growth with a single isomer of LAS [3-(4-sulfophenyl)dodecane] was concomitant with excretion of 4-(4-sulfophenyl)hexanoate, which was identified by matrix-assisted laser desorption ionization mass spectrometry. The growth yield (6.4 g of protein/mol of C) indicated mass balance, which, with the specific growth rate (0.05 h(-1)), indicated a specific utilization rate of LAS of 2.2 mkat/kg of protein. PMID- 10788360 TI - Products from the incomplete metabolism of pyrene by polycyclic aromatic hydrocarbon-degrading bacteria. AB - Pyrene is a regulated pollutant at sites contaminated with polycyclic aromatic hydrocarbons (PAH). It is mineralized by some bacteria but is also transformed to nonmineral products by a variety of other PAH-degrading bacteria. We examined the formation of such products by four bacterial strains and identified and further characterized the most apparently significant of these metabolites. Pseudomonas stutzeri strain P16 and Bacillus cereus strain P21 transformed pyrene primarily to cis-4,5-dihydro-4,5-dihydroxypyrene (PYRdHD), the first intermediate in the known pathway for aerobic bacterial mineralization of pyrene. Sphingomonas yanoikuyae strain R1 transformed pyrene to PYRdHD and pyrene-4,5-dione (PYRQ). Both strain R1 and Pseudomonas saccharophila strain P15 transform PYRdHD to PYRQ nearly stoichiometrically, suggesting that PYRQ is formed by oxidation of PYRdHD to 4,5-dihydroxypyrene and subsequent autoxidation of this metabolite. A pyrene mineralizing organism, Mycobacterium strain PYR-1, also transforms PYRdHD to PYRQ at high initial concentrations of PYRdHD. However, strain PYR-1 is able to use both PYRdHD and PYRQ as growth substrates. PYRdHD strongly inhibited phenanthrene degradation by strains P15 and R1 but had only a minor effect on strains P16 and P21. At their aqueous saturation concentrations, both PYRdHD and PYRQ severely inhibited benzo[a]pyrene mineralization by strains P15 and R1. Collectively, these findings suggest that products derived from pyrene transformation have the potential to accumulate in PAH-contaminated systems and that such products can significantly influence the removal of other PAH. However, these products may be susceptible to subsequent degradation by organisms able to metabolize pyrene more extensively if such organisms are present in the system. PMID- 10788361 TI - Mutagenesis of asp-569 of glucosyltransferase I glucansucrase modulates glucan and oligosaccharide synthesis. AB - Glucansucrases of oral streptococci and Leuconostoc mesenteroides are enzymes of medical and biotechnological interest that synthesize alpha-glucans. They can also synthesize oligosaccharides in the presence of a sugar acceptor. Previous reports have identified an amino acid residue that may affect the structure of the glucan product; therefore, random mutagenesis of the corresponding Asp-569 of Streptococcus downei glucosyltransferase I (GTF-I) was used to further understanding of its involvement in the catalytic mechanism and to evaluate how different amino acids can modulate glucan and oligosaccharide synthesis. GTF-I variants were obtained where Asp-569 was replaced by each of the different possible classes of amino acids. These were expressed in Escherichia coli and purified by means of a His(6) tag. The results showed that the amino acid in position 569 influences the structure of the glucan and the size of the oligosaccharides produced by GTF-I. The results suggest that the amino acid occupying this position is more likely to interact with the acceptor molecules (oligosaccharides or elongating glucan chain) than to be directly involved in glucosyl transfer from sucrose. Engineering of the equivalent position in glucansucrases thus appears to be a good target to expand the range of oligosaccharides synthesized. PMID- 10788362 TI - Antimicrobial peptides protect coho salmon from Vibrio anguillarum infections. AB - Fish losses from infectious diseases are a significant problem in aquaculture worldwide. Therefore, we investigated the ability of cationic antimicrobial peptides to protect against infection caused by the fish pathogen Vibrio anguillarum. To identify effective peptides for fish, the MICs of certain antimicrobial peptides against fish pathogens were determined in vitro. Two of the most effective antimicrobial peptides, CEME, a cecropin-melittin hybrid peptide, and pleurocidin amide, a C-terminally amidated form of the natural flounder peptide, were selected for in vivo studies. A single intraperitoneal injection of CEME did not affect mortality rates in juvenile coho salmon infected with V. anguillarum, the causative agent of vibriosis. Therefore, the peptides were delivered continuously using miniosmotic pumps placed in the peritoneal cavity. Twelve days after pump implantation, the fish received intraperitoneal injections of V. anguillarum at a dose that would kill 50 to 90% of the population. Fish receiving 200 microg of CEME per day survived longer and had significantly lower accumulated mortalities (13%) than the control groups (50 to 58%). Fish receiving pleurocidin amide at 250 microg per day also survived longer and had significantly lower accumulated mortalities (5%) than the control groups (67 to 75%). This clearly shows the potential for antimicrobial peptides to protect fish against infections and indicates that the strategy of overexpressing the peptides in transgenic fish may provide a method of decreasing bacterial disease problems. PMID- 10788363 TI - Detection and identification of Leishmania DNA within naturally infected sand flies by seminested PCR on minicircle kinetoplastic DNA. AB - A seminested PCR assay was developed in order to amplify the kinetoplast minicircle of Leishmania species from individual sand flies. The kinetoplast minicircle is an ideal target because it is present in 10,000 copies per cell and its sequence is known for most Leishmania species. The two-step PCR is carried out in a single tube using three primers, which were designed within the conserved area of the minicircle and contain conserved sequence blocks. The assay was able to detect as few as 3 parasites per individual sand fly and to amplify minicircle DNA from at least eight Leishmania species. This technique permits the processing of a large number of samples synchronously, as required for epidemiological studies, in order to study infection rates in sand fly populations and to identify potential insect vectors. Comparison of the sequences obtained from sand flies and mammal hosts will be crucial for developing hypotheses about the transmission cycles of Leishmania spp. in areas of endemicity. PMID- 10788364 TI - Genotypic and phenotypic diversity of phlD-containing Pseudomonas strains isolated from the rhizosphere of wheat. AB - Production of 2,4-diacetylphloroglucinol (2,4-DAPG) in the rhizosphere by strains of fluorescent Pseudomonas spp. results in the suppression of root diseases caused by certain fungal plant pathogens. In this study, fluorescent Pseudomonas strains containing phlD, which is directly involved in the biosynthesis of 2,4 DAPG, were isolated from the rhizosphere of wheat grown in soils from wheat growing regions of the United States and The Netherlands. To assess the genotypic and phenotypic diversity present in this collection, 138 isolates were compared to 4 previously described 2, 4-DAPG producers. Thirteen distinct genotypes, one of which represented over 30% of the isolates, were differentiated by whole-cell BOX-PCR. Representatives of this group were isolated from eight different soils taken from four different geographic locations. ERIC-PCR gave similar results overall, differentiating 15 distinct genotypes among all of the isolates. In most cases, a single genotype predominated among isolates obtained from each soil. Thirty isolates, representing all of the distinct genotypes and geographic locations, were further characterized. Restriction analysis of amplified 16S rRNA gene sequences revealed only three distinct phylogenetic groups, one of which accounted for 87% of the isolates. Phenotypic analyses based on carbon source utilization profiles revealed that all of the strains utilized 49 substrates and were unable to grow on 12 others. Individually, strains could utilize about two thirds of the 95 substrates present in Biolog SF-N plates. Multivariate analyses of utilization profiles revealed phenotypic groupings consistent with those defined by the genotypic analyses. PMID- 10788365 TI - A novel amidase (half-amidase) for half-amide hydrolysis involved in the bacterial metabolism of cyclic imides. AB - A novel amidase involved in bacterial cyclic imide metabolism was purified from Blastobacter sp. strain A17p-4. The enzyme physiologically functions in the second step of cyclic imide degradation, i.e., the hydrolysis of monoamidated dicarboxylates (half-amides) to dicarboxylates and ammonia. Enzyme production was enhanced by cyclic imides such as succinimide and glutarimide but not by amide compounds which are conventional substrates and inducers of known amidases. The purified amidase showed high catalytic efficiency toward half-amides such as succinamic acid (K(m) = 6.2 mM; k(cat) = 5.76 s(-1)) and glutaramic acid (K(m) = 2.8 mM; k(cat) = 2.23 s(-1)). However, the substrates of known amidases such as short-chain (C(2) to C(4)) aliphatic amides, long-chain (above C(16)) aliphatic amides, amino acid amides, aliphatic diamides, alpha-keto acid amides, N carbamoyl amino acids, and aliphatic ureides were not substrates for the enzyme. Based on its high specificity toward half-amides, the enzyme was named half amidase. This half-amidase exists as a monomer with an M(r) of 48,000 and was strongly inhibited by heavy metal ions and sulfhydryl reagents. PMID- 10788366 TI - Cell wall chemical composition of Enterococcus faecalis in the viable but nonculturable state. AB - The viable but nonculturable (VBNC) state is a survival mechanism adopted by many bacteria (including those of medical interest) when exposed to adverse environmental conditions. In this state bacteria lose the ability to grow in bacteriological media but maintain viability and pathogenicity and sometimes are able to revert to regular division upon restoration of normal growth conditions. The aim of this work was to analyze the biochemical composition of the cell wall of Enterococcus faecalis in the VBNC state in comparison with exponentially growing and stationary cells. VBNC enterococcal cells appeared as slightly elongated and were endowed with a wall more resistant to mechanical disruption than dividing cells. Analysis of the peptidoglycan chemical composition showed an increase in total cross-linking, which rose from 39% in growing cells to 48% in VBNC cells. This increase was detected in oligomers of a higher order than dimers, such as trimers (24% increase), tetramers (37% increase), pentamers (65% increase), and higher oligomers (95% increase). Changes were also observed in penicillin binding proteins (PBPs), the enzymes involved in the terminal stages of peptidoglycan assembly, with PBPs 5 and 1 being prevalent, and in autolytic enzymes, with a threefold increase in the activity of latent muramidase-1 in E. faecalis in the VBNC state. Accessory wall polymers such as teichoic acid and lipoteichoic acid proved unchanged and doubled in quantity, respectively, in VBNC cells in comparison to dividing cells. It is suggested that all these changes in the cell wall of VBNC enterococci are specific to this particular physiological state. This may provide indirect confirmation of the viability of these cells. PMID- 10788367 TI - Diversity and distribution of DNA sequences with affinity to ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in the Arctic Ocean. AB - The spatial distribution and diversity of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria (hereinafter referred to as ammonia oxidizers) in the Arctic Ocean were determined. The presence of ammonia oxidizers was detected by PCR amplification of 16S rRNA genes using a primer set specific for this group of organisms (nitA and nitB, which amplifies a 1.1-kb fragment between positions 137 and 1234, corresponding to Escherichia coli 16S rDNA numbering). We analyzed 246 samples collected from the upper water column (5 to 235 m) during March and April 1995, September and October 1996, and September 1997. Ammonia oxidizers were detected in 25% of the samples from 5 m, 80% of the samples from 55 m, 88% of the samples from 133 m, and 50% of the samples from 235 m. Analysis of nitA-nitB PCR product by nested PCR-denaturing gradient gel electrophoresis (DGGE) showed that all positive samples contained the same major band (band A), indicating the presence of a dominant, ubiquitous ammonia oxidizer in the Arctic Ocean basin. Twenty-two percent of the samples contained additional major bands. These samples were restricted to the Chukchi Sea shelf break, the Chukchi cap, and the Canada basin; areas likely influenced by Pacific inflow. The nucleotide sequence of the 1.1-kb nitA-nitB PCR product from a sample that contained only band A grouped with sequences designated group 1 marine Nitrosospira-like sequences. PCR-DGGE analysis of 122 clones from four libraries revealed that 67 to 71% of the inserts contained sequences with the same mobility as band A. Nucleotide sequences (1.1 kb) of another distinct group of clones, found only in 1995 samples (25%), fell into the group 5 marine Nitrosomonas-like sequences. Our results suggest that the Arctic Ocean beta-proteobacterial ammonia oxidizers have low diversity and are dominated by marine Nitrosospira-like organisms. Diversity appears to be higher in Western Arctic Ocean regions influenced by inflow from the Pacific Ocean through the Bering and Chukchi seas. PMID- 10788368 TI - Redirection of the respiro-fermentative flux distribution in Saccharomyces cerevisiae by overexpression of the transcription factor Hap4p. AB - Reduction of aerobic fermentation on sugars by altering the fermentative/oxidative balance is of significant interest for optimization of industrial production of Saccharomyces cerevisiae. Glucose control of oxidative metabolism in baker's yeast is partly mediated through transcriptional regulation of the Hap4p subunit of the Hap2/3/4/5p transcriptional activator complex. To alleviate glucose repression of oxidative metabolism, we constructed a yeast strain with constitutively elevated levels of Hap4p. Genetic analysis of expression levels of glucose-repressed genes and analysis of respiratory capacity showed that Hap4p overexpression (partly) relieves glucose repression of respiration. Analysis of the physiological properties of the Hap4p overproducer in batch cultures in fermentors (aerobic, glucose excess) has shown that the metabolism of this strain is more oxidative than in the wild-type strain, resulting in a significant reduced ethanol production and improvement of growth rate and a 40% gain in biomass yield. Our results show that modification of one or more transcriptional regulators can be a powerful and a widely applicable tool for redirection of metabolic fluxes in microorganisms. PMID- 10788370 TI - Horizontal heterogeneity of denitrifying bacterial communities in marine sediments by terminal restriction fragment length polymorphism analysis. AB - Although it is widely believed that horizontal patchiness exists in microbial sediment communities, determining the extent of variability or the particular members of the bacterial community which account for the observed differences among sites at various scales has not been routinely demonstrated. In this study, horizontal heterogeneity was examined in time and space for denitrifying bacteria in continental shelf sediments off Tuckerton, N.J., at the Rutgers University Long-Term Ecosystem Observatory (LEO-15). Characterization of the denitrifying community was done using PCR amplification of the nitrous oxide reductase (nosZ) gene combined with terminal restriction fragment length polymorphism analysis. Spatial scales from centimeters to kilometers were examined, while temporal variation was assayed over the course of 1995 to 1996. Sorenson's indices (pairwise similarity values) were calculated to permit comparison between samples. The similarities of benthic denitrifiers ranged from 0.80 to 0.85 for centimeter scale comparisons, from 0.52 to 0.79 for meter level comparisons, and from 0.23 to 0.53 for kilometer scale comparisons. Sorenson's indices for temporal comparisons varied from 0.12 to 0.74. A cluster analysis of the similarity values indicated that the composition of the denitrifier assemblages varied most significantly at the kilometer scale and between seasons at individual stations. Specific nosZ genes were identified which varied at centimeter, meter, or kilometer scales and may be associated with variability in meio- or macrofaunal abundance (centimeter scale), bottom topography (meter scale), or sediment characteristics (kilometer scale). PMID- 10788369 TI - Thermostabilization of proteins by diglycerol phosphate, a new compatible solute from the hyperthermophile Archaeoglobus fulgidus. AB - Diglycerol phosphate accumulates under salt stress in the archaeon Archaeoglobus fulgidus (L. O. Martins, R. Huber, H. Huber, K. O. Stetter, M. S. da Costa, and H. Santos, Appl. Environ. Microbiol. 63:896-902, 1997). This solute was purified after extraction from the cell biomass. In addition, the optically active and the optically inactive (racemic) forms of the compound were synthesized, and the ability of the solute to act as a protecting agent against heating was tested on several proteins derived from mesophilic or hyperthermophilic sources. Diglycerol phosphate exerted a considerable stabilizing effect against heat inactivation of rabbit muscle lactate dehydrogenase, baker's yeast alcohol dehydrogenase, and Thermococcus litoralis glutamate dehydrogenase. Highly homologous and structurally well-characterized rubredoxins from Desulfovibrio gigas, Desulfovibrio desulfuricans (ATCC 27774), and Clostridium pasteurianum were also examined for their thermal stabilities in the presence or absence of diglycerol phosphate, glycerol, and inorganic phosphate. These proteins showed different intrinsic thermostabilities, with half-lives in the range of 30 to 100 min. Diglycerol phosphate exerted a strong protecting effect, with approximately a fourfold increase in the half-lives for the loss of the visible spectra of D. gigas and C. pasteurianum rubredoxins. In contrast, the stability of D. desulfuricans rubredoxin was not affected. These different behaviors are discussed in the light of the known structural features of rubredoxins. The data show that diglycerol phosphate is a potentially useful protein stabilizer in biotechnological applications. PMID- 10788371 TI - Importance of passive diffusion in the uptake of polychlorinated biphenyls by phagotrophic protozoa. AB - Unicellular protozoan grazers represent a size class of organisms where a transition in the mechanism of chlorobiphenyl (CB) introduction, from diffusion through surface membranes to ingestion of contaminated prey, could occur. This study compares the relative importance of these two processes in the overall uptake of polychlorinated biphenyls by protists. Uptake rates and steady-state concentrations were compared in laboratory cultures of grazing and nongrazing protozoa. These experiments were conducted with a 10-microm marine scuticociliate (Uronema sp.), bacterial prey (Halomonas halodurans), and a suite of 21 CB congeners spanning a range of aqueous solubilities. The dominant pathway of CB uptake by both grazing and nongrazing protozoa was diffusion. Organic-carbon normalized CB concentrations (in the protozoan cell) were equivalent in grazing and nongrazing protozoa for all congeners studied. Rate constants for uptake into and loss from the protozoan cell were independently determined by using [3,3',4, 4'-(14)C]tetrachlorobiphenyl (IUPAC no. 77), 0.38 +/- 0.03 min(-1) and (1.1 +/- 0.1) x 10(-5) (g of organic carbon)(-1) min(-1), respectively. Magnitudes of the uptake and loss processes were calculated and compared by using a numerical model. The model result was consistent with data from the bioaccumulation experiment and supported the hypothesis that diffusive uptake is faster than ingestive uptake in phagotrophic unicellular protozoa. PMID- 10788372 TI - Fecal shedding of Campylobacter and Arcobacter spp. in dairy cattle. AB - Campylobacter jejuni, Campylobacter coli, and Arcobacter spp. were detected in feces of healthy dairy cows by highly specific multiplex-PCR assays. For C. jejuni, at this one-time sampling, cows from 80.6% of farm operations (n = 31) and 37.7% of individual dairy cattle fecal samples (n = 2,085) were positive. Farm management factors were correlated with prevalence in herds in which >25% of cows were positive for C. jejuni. Statistical significance was set at a P of 0.20. Using these criteria, application of manure with broadcast spreaders (P = 0.17), feeding of whole cottonseed or hulls (P = 0.17) or alfalfa (P = 0.15), and accessibility of feed to birds (P = 0.17) were identified as possible risk factors for C. jejuni infection. C. coli was detected in at least one animal in 19.4% of operations and 1.8% of individual cows (n = 2,085). At the herd level, use of broadcaster spreaders was not a risk factor for C. coli infection. For Arcobacter, cows from 71% of dairy operations (n = 31) and 14.3% of individual dairy cattle fecal samples (n = 1,682) were positive. At the herd level, for Arcobacter spp., feeding of alfalfa (P = 0.11) and use of individual waterers (P = 0.19) were protective. This is the first description of Arcobacter spp. in clinically healthy dairy cattle and the first attempt to correlate their presence with C. jejuni. PMID- 10788373 TI - Lactic acid permeabilizes gram-negative bacteria by disrupting the outer membrane. AB - The effect of lactic acid on the outer membrane permeability of Escherichia coli O157:H7, Pseudomonas aeruginosa, and Salmonella enterica serovar Typhimurium was studied utilizing a fluorescent-probe uptake assay and sensitization to bacteriolysis. For control purposes, similar assays were performed with EDTA (a permeabilizer acting by chelation) and with hydrochloric acid, the latter at pH values corresponding to those yielded by lactic acid, and also in the presence of KCN. Already 5 mM (pH 4.0) lactic acid caused prominent permeabilization in each species, the effect in the fluorescence assay being stronger than that of EDTA or HCl. Similar results were obtained in the presence of KCN, except for P. aeruginosa, for which an increase in the effect of HCl was observed in the presence of KCN. The permeabilization by lactic and hydrochloric acid was partly abolished by MgCl(2). Lactic acid sensitized E. coli and serovar Typhimurium to the lytic action of sodium dodecyl sulfate (SDS) more efficiently than did HCl, whereas both acids sensitized P. aeruginosa to SDS and to Triton X-100. P. aeruginosa was effectively sensitized to lysozyme by lactic acid and by HCl. Considerable proportions of lipopolysaccharide were liberated from serovar Typhimurium by these acids; analysis of liberated material by electrophoresis and by fatty acid analysis showed that lactic acid was more active than EDTA or HCl in liberating lipopolysaccharide from the outer membrane. Thus, lactic acid, in addition to its antimicrobial property due to the lowering of the pH, also functions as a permeabilizer of the gram-negative bacterial outer membrane and may act as a potentiator of the effects of other antimicrobial substances. PMID- 10788374 TI - Reduction of Fe(III), Cr(VI), U(VI), and Tc(VII) by Deinococcus radiodurans R1. AB - Deinococcus radiodurans is an exceptionally radiation-resistant microorganism capable of surviving acute exposures to ionizing radiation doses of 15,000 Gy and previously described as having a strictly aerobic respiratory metabolism. Under strict anaerobic conditions, D. radiodurans R1 reduced Fe(III)-nitrilotriacetic acid coupled to the oxidation of lactate to CO(2) and acetate but was unable to link this process to growth. D. radiodurans reduced the humic acid analog anthraquinone-2,6-disulfonate (AQDS) to its dihydroquinone form, AH(2)DS, which subsequently transferred electrons to the Fe(III) oxides hydrous ferric oxide and goethite via a previously described electron shuttle mechanism. D. radiodurans reduced the solid-phase Fe(III) oxides in the presence of either 0.1 mM AQDS or leonardite humic acids (2 mg ml(-1)) but not in their absence. D. radiodurans also reduced U(VI) and Tc(VII) in the presence of AQDS. In contrast, Cr(VI) was directly reduced in anaerobic cultures with lactate although the rate of reduction was higher in the presence of AQDS. The results are the first evidence that D. radiodurans can reduce Fe(III) coupled to the oxidation of lactate or other organic compounds. Also, D. radiodurans, in combination with humic acids or synthetic electron shuttle agents, can reduce U and Tc and thus has potential applications for remediation of metal- and radionuclide-contaminated sites where ionizing radiation or other DNA-damaging agents may restrict the activity of more sensitive organisms. PMID- 10788375 TI - Interactions between pyruvate and lactate metabolism in Propionibacterium freudenreichii subsp. shermanii: in vivo (13)C nuclear magnetic resonance studies. AB - In vivo (13)C nuclear magnetic resonance spectroscopy was used to elucidate the pathways and the regulation of pyruvate metabolism and pyruvate-lactate cometabolism noninvasively in living-cell suspensions of Propionibacterium freudenreichii subsp. shermanii. The most important result of this work concerns the modification of fluxes of pyruvate metabolism induced by the presence of lactate. Pyruvate was temporarily converted to lactate and alanine; the flux to acetate synthesis was maintained, but the flux to propionate synthesis was increased; and the reverse flux of the first part of the Wood-Werkman cycle, up to acetate synthesis, was decreased. Pyruvate was consumed at apparent initial rates of 148 and 90 micromol. min(-1). g(-1) (cell dry weight) when it was the sole substrate or cometabolized with lactate, respectively. Lactate was consumed at an apparent initial rate of 157 micromol. min(-1). g(-1) when it was cometabolized with pyruvate. P. shermanii used several pathways, namely, the Wood Werkman cycle, synthesis of acetate and CO(2), succinate synthesis, gluconeogenesis, the tricarboxylic acid cycle, and alanine synthesis, to manage its pyruvate pool sharply. In both types of experiments, acetate synthesis and the Wood-Werkman cycle were the metabolic pathways used most. PMID- 10788376 TI - Structural changes and interactions involved in the Ca(2+)-triggered stabilization of the cell-bound cell envelope proteinase in Lactococcus lactis subsp. cremoris SK11. AB - The cell-bound cell envelope proteinase (CEP) of the mesophilic cheese-starter organism Lactococcus lactis subsp. cremoris SK11 is protected from rapid thermal inactivation at 25 degrees C by calcium bound to weak binding sites. The interactions with calcium are believed to trigger reversible structural rearrangements which are coupled with changes in specific activity (F. A. Exterkate and A. C. Alting, Appl. Env. Microbiol. 65:1390-1396, 1999). In order to determine the significance of the rearrangements for CEP stability and the nature of the interactions involved, the effects of the net charge present on the enzyme and of different neutral salts were studied with the stable Ca-loaded CEP, the unstable so-called "Ca-free" CEP and with the Ca-free CEP which was stabilized nonspecifically and essentially in its native conformation by the nonionic additive sucrose. The results suggest that strengthening of hydrophobic interactions is conducive to stabilization of the Ca-free CEP. On the other hand, a hydrophobic effect contributes significantly to the stability of the Ca-loaded CEP; a phased salting-in effect by a chaotropic salt suggests a complex inactivation process of this enzyme due to weakening of hydrophobic interactions and involving an intermediate enzyme species. Moreover, a Ca-triggered increase of a relatively significant hydrophobic effect in the sucrose-stabilized Ca-free CEP occurs. It is suggested that in the Ca-free CEP the absence of both local calcium-mediated backbone rigidification and neutralization of negative electrostatic potentials in the weak Ca-binding sites, and in addition the lack of significant hydrophobic stabilization, increase the relative effectiveness of electrostatic repulsive forces on the protein to an extent that causes the observed instability. The conditions in cheese seem to confer stability upon the cell-bound enzyme; its possible involvement in proteolysis throughout the ripening period is discussed. PMID- 10788377 TI - Targeted disruption of the kstD gene encoding a 3-ketosteroid delta(1) dehydrogenase isoenzyme of Rhodococcus erythropolis strain SQ1. AB - Microbial phytosterol degradation is accompanied by the formation of steroid pathway intermediates, which are potential precursors in the synthesis of bioactive steroids. Degradation of these steroid intermediates is initiated by Delta(1)-dehydrogenation of the steroid ring structure. Characterization of a 2.9 kb DNA fragment of Rhodococcus erythropolis SQ1 revealed an open reading frame (kstD) showing similarity with known 3-ketosteroid Delta(1)-dehydrogenase genes. Heterologous expression of kstD yielded 3-ketosteroid Delta(1)-dehydrogenase (KSTD) activity under the control of the lac promoter in Escherichia coli. Targeted disruption of the kstD gene in R. erythropolis SQ1 was achieved, resulting in loss of more than 99% of the KSTD activity. However, growth on the steroid substrate 4-androstene-3,17-dione or 9alpha-hydroxy-4-androstene-3,17 dione was not abolished by the kstD gene disruption. Bioconversion of phytosterols was also not blocked at the level of Delta(1)-dehydrogenation in the kstD mutant strain, since no accumulation of steroid pathway intermediates was observed. Thus, inactivation of kstD is not sufficient for inactivation of the Delta(1)-dehydrogenase activity. Native polyacrylamide gel electrophoresis of cell extracts stained for KSTD activity showed that R. erythropolis SQ1 in fact harbors two activity bands, one of which is absent in the kstD mutant strain. PMID- 10788378 TI - Physiological responses to starvation in the marine oligotrophic ultramicrobacterium Sphingomonas sp. strain RB2256. AB - Sphingomonas sp. strain RB2256 is representative of the ultramicrobacteria that proliferate in oligotrophic marine waters. While this class of bacteria is well adapted for growth with low concentrations of nutrients, their ability to respond to complete nutrient deprivation has not previously been investigated. In this study, we examined two-dimensional protein profiles for logarithmic and stationary-phase cells and found that protein spot intensity was regulated by up to 70-fold. A total of 72 and 177 spots showed increased or decreased intensity, respectively, by at least twofold during starvation. The large number of protein spots (1,500) relative to the small genome size (ca. 1.5 Mb) indicates that gene expression may involve co- and posttranslational modifications of proteins. Rates of protein and RNA synthesis were examined throughout the growth phase and up to 7 days of starvation and revealed that synthesis was highly regulated. Rates of protein synthesis and cellular protein content were compared to ribosome content, demonstrating that ribosome synthesis was not directly linked to protein synthesis and that the function of ribosomes may not be limited to translation. By comparing the genetic capacity and physiological responses to starvation of RB2256 to those of the copiotrophic marine bacterium Vibrio angustum S14 (J. Ostling, L. Holmquist, and S. Kjelleberg, J. Bacteriol. 178:4901-4908, 1996), the characteristics of a distinct starvation response were defined for Sphingomonas strain RB2256. The capacity of this ultramicrobacterium to respond to starvation is discussed in terms of the ecological relevance of complete nutrient deprivation in an oligotrophic marine environment. These studies provide the first evidence that marine oligotrophic ultramicrobacteria may be expected to include a starvation response and the capacity for a high degree of gene regulation. PMID- 10788379 TI - In vitro ATP regeneration from polyphosphate and AMP by polyphosphate:AMP phosphotransferase and adenylate kinase from Acinetobacter johnsonii 210A. AB - In vitro enzyme-based ATP regeneration systems are important for improving yields of ATP-dependent enzymatic reactions for preparative organic synthesis and biocatalysis. Several enzymatic ATP regeneration systems have been described but have some disadvantages. We report here on the use of polyphosphate:AMP phosphotransferase (PPT) from Acinetobacter johnsonii strain 210A in an ATP regeneration system based on the use of polyphosphate (polyP) and AMP as substrates. We have examined the substrate specificity of PPT and demonstrated ATP regeneration from AMP and polyP using firefly luciferase and hexokinase as model ATP-requiring enzymes. PPT catalyzes the reaction polyP(n) + AMP --> ADP + polyP(n-1). The ADP can be converted to ATP by adenylate kinase (AdK). Substrate specificity with nucleoside and 2'-deoxynucleoside monophosphates was examined using partially purified PPT by measuring the formation of nucleoside diphosphates with high-pressure liquid chromatography. AMP and 2'-dAMP were efficiently phosphorylated to ADP and 2'-dADP, respectively. GMP, UMP, CMP, and IMP were not converted to the corresponding diphosphates at significant rates. Sufficient AdK and PPT activity in A. johnsonii 210A cell extract allowed demonstration of polyP-dependent ATP regeneration using a firefly luciferase based ATP assay. Bioluminescence from the luciferase reaction, which normally decays very rapidly, was sustained in the presence of A. johnsonii 210A cell extract, MgCl(2), polyP(n=35), and AMP. Similar reaction mixtures containing strain 210A cell extract or partially purified PPT, polyP, AMP, glucose, and hexokinase formed glucose 6-phosphate. The results indicate that PPT from A. johnsonii is specific for AMP and 2'-dAMP and catalyzes a key reaction in the cell-free regeneration of ATP from AMP and polyP. The PPT/AdK system provides an alternative to existing enzymatic ATP regeneration systems in which phosphoenolpyruvate and acetylphosphate serve as phosphoryl donors and has the advantage that AMP and polyP are stabile, inexpensive substrates. PMID- 10788380 TI - Redox chemistry in laccase-catalyzed oxidation of N-hydroxy compounds. AB - 1-Hydroxybenzotriazole, violuric acid, and N-hydroxyacetanilide are three N-OH compounds capable of mediating a range of laccase-catalyzed biotransformations, such as paper pulp delignification and degradation of polycyclic hydrocarbons. The mechanism of their enzymatic oxidation was studied with seven fungal laccases. The oxidation had a bell-shaped pH-activity profile with an optimal pH ranging from 4 to 7. The oxidation rate was found to be dependent on the redox potential difference between the N-OH substrate and laccase. A laccase with a higher redox potential or an N-OH compound with a lower redox potential tended to have a higher oxidation rate. Similar to the enzymatic oxidation of phenols, phenoxazines, phenothiazines, and other redox-active compounds, an "outer-sphere" type of single-electron transfer from the substrate to laccase and proton release are speculated to be involved in the rate-limiting step for N-OH oxidation. PMID- 10788381 TI - Mitotic recombination and genetic changes in Saccharomyces cerevisiae during wine fermentation. AB - Natural strains of Saccharomyces cerevisiae are prototrophic homothallic yeasts that sporulate poorly, are often heterozygous, and may be aneuploid. This genomic constitution may confer selective advantages in some environments. Different mechanisms of recombination, such as meiosis or mitotic rearrangement of chromosomes, have been proposed for wine strains. We studied the stability of the URA3 locus of a URA3/ura3 wine yeast in consecutive grape must fermentations. ura3/ura3 homozygotes were detected at a rate of 1 x 10(-5) to 3 x 10(-5) per generation, and mitotic rearrangements for chromosomes VIII and XII appeared after 30 mitotic divisions. We used the karyotype as a meiotic marker and determined that sporulation was not involved in this process. Thus, we propose a hypothesis for the genome changes in wine yeasts during vinification. This putative mechanism involves mitotic recombination between homologous sequences and does not necessarily imply meiosis. PMID- 10788382 TI - Altered regulation of 15-acetyldeoxynivalenol production in Fusarium graminearum. AB - Most Fusarium graminearum isolates produce low or undetectable levels of trichothecenes in liquid shake cultures, making it difficult to perform biochemical studies of trichothecene biosynthesis. To develop strains with higher levels of trichothecene production under liquid shake conditions we transformed F. graminearum with both a reporter gene containing a homologous trichothecene pathway gene promoter (TRI5) and a gene encoding a heterologous trichothecene pathway transcription factor (TRI6). The TRI5 and TRI6 genes are part of the trichothecene pathway gene clusters of both Fusarium sporotrichioides and F. graminearum. These genes encode trichodiene synthase (encoded by TRI5), the first enzyme in the trichothecene pathway, and a transcription factor (encoded by TRI6) required for pathway gene expression. Transformation of F. graminearum with plasmids containing either an F. graminearum TRI5 promoter fragment (FGTRI5(P)) or FGTRI5(P) coupled with the beta-D-glucuronidase (GUS) reporter gene resulted in the identification of several transformants capable of producing 45 to 200 mg of 15-acetyldeoxynivalenol (15-ADON)/liter in liquid shake culture after 7 days. Increased 15-ADON production was only observed in transformants where plasmid integration occurred through the FGTRI5(P) sequence and was not accompanied by increased GUS expression. 15-ADON production was further increased in liquid culture up to 1,200 mg/liter following introduction of the F. sporotrichioides TRI6 gene (FSTRI16) into F. graminearum. The effects of FSTRI6 on 15-ADON production also depended on plasmid integration via homologous recombination of the FGTRI5(P) fragment and resulted in a 100-fold increase in GUS expression. High-level production of 15-ADON in liquid shake cultures provides a convenient method for large-scale trichothecene preparation. The results suggest that targeting transformation vector integration to FGTRI5(P) alters pathway gene expression and are consistent with the proposed conservation of TRI6 function between Fusarium species. PMID- 10788383 TI - Comparison of methods for detection of Erysipelothrix spp. and their distribution in some Australasian seafoods. AB - For many years, Erysipelothrix rhusiopathiae has been known to be the causative agent of the occupationally related infection erysipeloid. A survey of the distribution of Erysipelothrix spp. in 19 Australasian seafoods was conducted, and methodologies for the detection of Erysipelothrix spp. were evaluated. Twenty one Erysipelothrix spp. were isolated from 52 seafood parts. Primary isolation of Erysipelothrix spp. was most efficiently achieved with brain heart infusion broth enrichment followed by subculture onto a selective brain heart infusion agar containing kanamycin, neomycin, and vancomycin after 48 h of incubation. Selective tryptic soy broth, with 48 h of incubation, was the best culture method for the detection of Erysipelothrix spp. with PCR. PCR detection was 50% more sensitive than culture. E. rhusiopathiae was isolated from a variety of different fish, cephalopods, and crustaceans, including a Western rock lobster (Panulirus cygnus). There was no significant correlation between the origin of the seafoods tested and the distribution of E. rhusiopathiae. An organism indistinguishable from Erysipelothrix tonsillarum was isolated for the first time from an Australian oyster and a silver bream. Overall, Erysipelothrix spp. were widely distributed in Australasian seafoods, illustrating the potential for erysipeloid like infections in fishermen. PMID- 10788384 TI - Development of a highly sensitive nested-PCR procedure using a single closed tube for detection of Erwinia amylovora in asymptomatic plant material. AB - A novel method, which involves a nested PCR in a single closed tube, was developed for the sensitive detection of Erwinia amylovora in plant material. The external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific DNA fragment from plasmid pEA29. The procedure involved two consecutive PCRs, the first of which was performed at a higher annealing temperature that allowed amplification only by the external primer pair. Using pure cultures of E. amylovora, the sensitivity of the nested PCR in one tube was similar to that of a standard nested PCR in two tubes. The specificity and sensitivity were greater than those of standard PCR procedures that used a single primer pair. The presence of inhibitors in plant material, very common in E. amylovora hosts, is overcome with this system in combination with a simple DNA extraction protocol because it eliminates many of the inhibitory compounds. In addition, it needs a very small sample volume (1 microl of DNA extracted). With 83 samples of naturally infected material, this method achieved better results than any other PCR technique: standard PCR detected 55% of positive samples, two-tube nested PCR detected 71% of positive samples, and nested PCR in a single closed tube detected 78% of positive samples. When analyzing asymptomatic plant material, the number of positive samples detected by the developed nested PCR was also the highest, compared with the PCR protocols indicated previously (17, 20, and 25% of 251 samples analyzed, respectively). This method is proposed for the detection of endophytic and epiphytic populations of E. amylovora in epidemiological studies and for routine use in quarantine surveys, due to its high sensitivity, specificity, speed, and simplicity. PMID- 10788385 TI - Inhibition of luminescence and virulence in the black tiger prawn (Penaeus monodon) pathogen Vibrio harveyi by intercellular signal antagonists. AB - Expression of luminescence in the Penaeus monodon pathogen Vibrio harveyi is regulated by an intercellular quorum sensing mechanism involving the synthesis and detection of two signaling molecules, one of which is N-hydroxy butanoyl-L homoserine lactone and the other of which is uncharacterized. Indirect evidence has suggested that virulence, associated with a toxic extracellular protein, and luminescence in V. harveyi are coregulated. In this study the effects of an acylated homoserine lactone antagonist produced by the marine alga Delisea pulchra on luminescence and toxin production in a virulent strain of V. harveyi were analyzed. Luminescence and toxin production were both inhibited by the signal antagonist at concentrations that had no impact on growth. Toxin production was found to be prematurely induced in V. harveyi cultures incubated in a 10% conditioned medium. Additionally, a significant reduction in the toxicity of concentrated supernatant extracts from V. harveyi cultures incubated in the presence of the signal antagonist, as measured by in vivo toxicity assays in mice and prawns, was observed. These results suggest that intercellular signaling antagonists have potential utility in the control of V. harveyi prawn infections. PMID- 10788386 TI - Molecular and phenotypic characterization of Pseudomonas spp. isolated from milk. AB - Putative Pseudomonas spp. isolated predominantly from raw and processed milk were characterized by automated ribotyping and by biochemical reactions. Isolates were biochemically profiled using the Biolog system and API 20 NE and by determining the production of proteases, lipases, and lecithinases for each isolate. Isolates grouped into five coherent clusters, predominated by the species P. putida (cluster A), P. fluorescens (cluster B), P. fragi (as identified by Biolog) or P. fluorescens (as identified by API 20 NE) (cluster C), P. fragi (as identified by Biolog) or P. putida (as identified by API 20 NE) (cluster D), and P. fluorescens (cluster E). Isolates within each cluster also displayed similar enzyme activities. Isolates in clusters A, C, and D were generally negative for all three enzyme activities; isolates in cluster B were predominantly positive for all three enzyme activities; and isolates in cluster E were negative for lecithinase but predominantly positive for protease and lipase activities. Thus, only isolates from clusters B and E produced enzyme activities associated with dairy product flavor defects. Thirty-eight ribogroups were differentiated among the 70 isolates. Ribotyping was highly discriminatory for dairy Pseudomonas isolates, with a Simpson's index of discrimination of 0.955. Isolates of the same ribotype were never classified into different clusters, and ribotypes within a given cluster generally showed similar ribotype patterns; thus, specific ribotype fragments may be useful markers for tracking the sources of pseudomonads in dairy production systems. Our results suggest that ribogroups are generally homogeneous with respect to nomenspecies and biovars, confirming the identification potential of ribotyping for Pseudomonas spp. PMID- 10788387 TI - Nitrite reductase genes (nirK and nirS) as functional markers to investigate diversity of denitrifying bacteria in pacific northwest marine sediment communities. AB - Genetic heterogeneity of denitrifying bacteria in sediment samples from Puget Sound and two sites on the Washington continental margin was studied by PCR approaches amplifying nirK and nirS genes. These structurally different but functionally equivalent single-copy genes coding for nitrite reductases, a key enzyme of the denitrification process, were used as a molecular marker for denitrifying bacteria. nirS sequences could be amplified from samples of both sampling sites, whereas nirK sequences were detected only in samples from the Washington margin. To assess the underlying nir gene structure, PCR products of both genes were cloned and screened by restriction fragment length polymorphism (RFLP). Rarefraction analysis revealed a high level of diversity especially for nirS clones from Puget Sound and a slightly lower level of diversity for nirK and nirS clones from the Washington margin. One group dominated within nirK clones, but no dominance and only a few redundant clones were seen between sediment samples for nirS clones in both habitats. Hybridization and sequencing confirmed that all but one of the 228 putative nirS clones were nirS with levels of nucleotide identities as low as 45.3%. Phylogenetic analysis grouped nirS clones into three distinct subclusters within the nirS gene tree which corresponded to the two habitats from which they were obtained. These sequences had little relationship to any strain with known nirS sequences or to isolates (mostly close relatives of Pseudomonas stutzeri) from the Washington margin sediment samples. nirK clones were more closely related to each other than were the nirS clones, with 78.6% and higher nucleotide identities; clones showing only weak hybridization signals were not related to known nirK sequences. All nirK clones were also grouped into a distinct cluster which could not be placed with any strain with known nirK sequences. These findings show a very high diversity of nir sequences within small samples and that these novel nir clusters, some very divergent from known sequences, are not known in cultivated denitrifiers. PMID- 10788388 TI - Identification and characterization of three differentially expressed genes, encoding S-adenosylhomocysteine hydrolase, methionine aminopeptidase, and a histone-like protein, in the toxic dinoflagellate Alexandrium fundyense. AB - Genes showing differential expression related to the early G(1) phase of the cell cycle during synchronized circadian growth of the toxic dinoflagellate Alexandrium fundyense were identified and characterized by differential display (DD). The determination in our previous work that toxin production in Alexandrium is relegated to a narrow time frame in early G(1) led to the hypothesis that transcriptionally up- or downregulated genes during this subphase of the cell cycle might be related to toxin biosynthesis. Three genes, encoding S adenosylhomocysteine hydrolase (Sahh), methionine aminopeptidase (Map), and a histone-like protein (HAf), were isolated. Sahh was downregulated, while Map and HAf were upregulated, during the early G(1) phase of the cell cycle. Sahh and Map encoded amino acid sequences with about 90 and 70% similarity to those encoded by several eukaryotic and prokaryotic Sahh and Map genes, respectively. The partial Map sequence also contained three cobalt binding motifs characteristic of all Map genes. HAf encoded an amino acid sequence with 60% similarity to those of two histone-like proteins from the dinoflagellate Crypthecodinium cohnii Biecheler. This study documents the potential of applying DD to the identification of genes that are related to physiological processes or cell cycle events in phytoplankton under conditions where small sample volumes represent an experimental constraint. The identification of an additional 21 genes with various cell cycle-related DD patterns also provides evidence for the importance of pretranslational or transcriptional regulation in dinoflagellates, contrary to previous reports suggesting the possibility that translational mechanisms are the primary means of circadian regulation in this group of organisms. PMID- 10788389 TI - Multiplex PCR for the detection of Lactobacillus pontis and two related species in a sourdough fermentation. AB - A specific multiplex PCR assay based on the amplification of parts of the 16S rRNA molecule was designed. Primers derived from variable regions of the 16S rRNA provided a means of easily differentiating the species Lactobacillus pontis and Lactobacillus panis. They could be clearly discriminated from the phylogenetically related species Lactobacillus vaginalis, Lactobacillus oris, and Lactobacillus reuteri and from other lactobacilli commonly known to be present in sourdough. Other strains isolated together with L. pontis from an industrial sourdough fermentation could be clearly separated from these species by comparative sequence analysis and construction of a specific PCR primer. For a fast identification a DNA isolation protocol based on the ultrasonic lysis of cells from single colonies was developed. To demonstrate the potential of such techniques for tracking these organisms in a laboratory-scale fermentation, we combined the specific PCR assay with direct DNA extraction from the organisms in the sourdough without previous cultivation. PMID- 10788390 TI - PhaG-mediated synthesis of Poly(3-hydroxyalkanoates) consisting of medium-chain length constituents from nonrelated carbon sources in recombinant Pseudomonas fragi. AB - Recently, a new metabolic link between fatty acid de novo biosynthesis and biosynthesis of poly(3-hydroxy-alkanoate) consisting of medium-chain-length constituents (C(6) to C(14)) (PHA(MCL)), catalyzed by the 3-hydroxydecanoyl-[acyl carrier-protein]:CoA transacylase (PhaG), has been identified in Pseudomonas putida (B. H. A. Rehm, N. Kruger, and A. Steinbuchel, J. Biol. Chem. 273:24044 24051, 1998). To establish this PHA-biosynthetic pathway in a non-PHA accumulating bacterium, we functionally coexpressed phaC1 (encoding PHA synthase 1) from Pseudomonas aeruginosa and phaG (encoding the transacylase) from P. putida in Pseudomonas fragi. The recombinant strains of P. fragi were cultivated on gluconate as the sole carbon source, and PHA accumulation to about 14% of the total cellular dry weight was achieved. The respective polyester was isolated, and GPC analysis revealed a weight average molar mass of about 130,000 g mol(-1) and a polydispersity of 2.2. The PHA was composed mainly (60 mol%) of 3 hydroxydecanoate. These data strongly suggested that functional expression of phaC1 and phaG established a new pathway for PHA(MCL) biosynthesis from nonrelated carbon sources in P. fragi. When fatty acids were used as the carbon source, no PHA accumulation was observed in PHA synthase-expressing P. fragi, whereas application of the beta-oxidation inhibitor acrylic acid mediated PHA(MCL) accumulation. The substrate for the PHA synthase PhaC1 is therefore presumably directly provided through the enzymatic activity of the transacylase PhaG by the conversion of (R)-3-hydroxydecanoyl-ACP to (R)-3-hydroxydecanoyl-CoA when the organism is cultivated on gluconate. Here we demonstrate for the first time the establishment of PHA(MCL) synthesis from nonrelated carbon sources in a non-PHA-accumulating bacterium, employing fatty acid de novo biosynthesis and the enzymes PhaG (a transacylase) and PhaC1 (a PHA synthase). PMID- 10788391 TI - Coexistence of two different O demethylation systems in lignin metabolism by Sphingomonas paucimobilis SYK-6: cloning and sequencing of the lignin biphenyl specific O-demethylase (LigX) gene. AB - Sphingomonas paucimobilis SYK-6 can grow on several dimeric model compounds of lignin as a carbon and energy source. It has O demethylation systems on three kinds of substrates: 5, 5'-dehydrodivanillic acid (DDVA), syringate, and vanillate. We previously reported the cloning of a gene involved in the tetrahydrofolate-dependent O demethylation of syringate and vanillate. In the study reported here, we cloned the gene responsible for DDVA O demethylation. Using nitrosoguanidine mutagenesis, a mutant strain, NT-1, which could not degrade DDVA but could degrade syringate and vanillate, was isolated and was used to clone the gene responsible for the O demethylation of DDVA by complementation. Sequencing analysis showed an open reading frame (designated ligX) of 1,266 bp in this fragment. The deduced amino acid sequence of LigX had similarity to class I type oxygenases. LigX was involved in O demethylation activity on DDVA but not on vanillate and syringate. DDVA O demethylation activity in S. paucimobilis SYK-6 cell extracts was inhibited by addition of the LigX polyclonal antiserum. Thus, LigX is an essential enzyme for DDVA O demethylation in SYK-6. S. paucimobilis SYK-6 has two O demethylation systems: one is an oxygenative demethylase system, and the other is a tetrahydrofolate-dependent methyltransferase system. PMID- 10788392 TI - Construction and evaluation of a novel bifunctional N-carbamylase-D-hydantoinase fusion enzyme. AB - A fully enzymatic process employing two sequential enzymes, D-hydantoinase and N carbamylase, is a typical case requiring combined enzyme activity for the production of D-amino acids. To test the possibility of generating a bifunctional fusion enzyme, we constructed a fusion protein via end-to-end fusion of a whole gene that encodes an intact protein at the N terminus of the D-hydantoinase. Firstly, maltose-binding protein (MBP) gene of E. coli was fused with D hydantoinase gene from Bacillus stearothermophilus SD1, and the properties of the resulting fusion protein (MBP-HYD) were compared with those of native D hydantoinase. Gel filtration and kinetic analyses clearly demonstrated that the typical characteristics of D-hydantoinase are maintained even in a fusion state. Based on this result, we constructed an artificial fusion enzyme composed of the whole length of N-carbamylase (304 amino acids [aa]) from Agrobacterim radiobacter NRRL B11291 and D-hydantoinase (471 aa). The fusion enzyme (CAB-HYD) was functionally expressed with an expected molecular mass of 86 kDa and efficiently converted exogenous hydantoin derivatives to the D-amino acids. A related D-hydantoinase (HYD1) gene from Bacillus thermocatenulatus GH2 was also fused with the N-carbamylase gene at its N terminus. The resulting enzyme (CAB HYD1) was bifunctional as expected and showed better performance than the CAB-HYD fusion enzyme. The conversion of hydantoin derivatives to corresponding amino acids by the fusion enzymes was much higher than that by the separately expressed enzymes, and comparable to that by the coexpressed enzymes. Thus, the fusion enzyme might be useful as a potential biocatalyst for the production of nonnatural amino acids. PMID- 10788393 TI - Aerobic degradation of dinitrotoluenes and pathway for bacterial degradation of 2,6-dinitrotoluene. AB - An oxidative pathway for the mineralization of 2,4-dinitrotoluene (2, 4-DNT) by Burkholderia sp. strain DNT has been reported previously. We report here the isolation of additional strains with the ability to mineralize 2,4-DNT by the same pathway and the isolation and characterization of bacterial strains that mineralize 2, 6-dinitrotoluene (2,6-DNT) by a different pathway. Burkholderia cepacia strain JS850 and Hydrogenophaga palleronii strain JS863 grew on 2,6-DNT as the sole source of carbon and nitrogen. The initial steps in the pathway for degradation of 2,6-DNT were determined by simultaneous induction, enzyme assays, and identification of metabolites through mass spectroscopy and nuclear magnetic resonance. 2,6-DNT was converted to 3-methyl-4-nitrocatechol by a dioxygenation reaction accompanied by the release of nitrite. 3-Methyl-4-nitrocatechol was the substrate for extradiol ring cleavage yielding 2-hydroxy-5-nitro-6-oxohepta-2,4 dienoic acid, which was converted to 2-hydroxy-5-nitropenta-2,4-dienoic acid. 2, 4-DNT-degrading strains also converted 2,6-DNT to 3-methyl-4-nitrocatechol but did not metabolize the 3-methyl-4-nitrocatechol. Although 2,6-DNT prevented the degradation of 2,4-DNT by 2,4-DNT-degrading strains, the effect was not the result of inhibition of 2,4-DNT dioxygenase by 2,6-DNT or of 4-methyl-5 nitrocatechol monooxygenase by 3-methyl-4-nitrocatechol. PMID- 10788394 TI - Identification of fluoropyrogallols as new intermediates in biotransformation of monofluorophenols in Rhodococcus opacus 1cp. AB - The transformation of monofluorophenols by whole cells of Rhodococcus opacus 1cp was investigated, with special emphasis on the nature of hydroxylated intermediates formed. Thin-layer chromatography, mass spectrum analysis, and (19)F nuclear magnetic resonance demonstrated the formation of fluorocatechol and trihydroxyfluorobenzene derivatives from each of three monofluorophenols. The (19)F chemical shifts and proton-coupled splitting patterns of the fluorine resonances of the trihydroxyfluorobenzene products established that the trihydroxylated aromatic metabolites contained hydroxyl substituents on three adjacent carbon atoms. Thus, formation of 1,2, 3-trihydroxy-4-fluorobenzene (4 fluoropyrogallol) from 2-fluorophenol and formation of 1,2,3-trihydroxy-5 fluorobenzene (5-fluoropyrogallol) from 3-fluorophenol and 4-fluorophenol were observed. These results indicate the involvement of fluoropyrogallols as previously unidentified metabolites in the biotransformation of monofluorophenols in R. opacus 1cp. PMID- 10788395 TI - Monitoring precursor 16S rRNAs of Acinetobacter spp. in activated sludge wastewater treatment systems. AB - Recently, Cangelosi and Brabant used oligonucleotide probes targeting the precursor 16S rRNA of Escherichia coli to demonstrate that the levels of precursor rRNA were more sensitive to changes in growth phase than the levels of total rRNA (G. A. Cangelosi and W. H. Brabant, J. Bacteriol. 179:4457-4463, 1997). In order to measure changes in the levels of precursor rRNA in activated sludge systems, we designed oligonucleotide probes targeting the 3' region of the precursor 16S rRNA of Acinetobacter spp. We used these probes to monitor changes in the level of precursor 16S rRNA during batch growth of Acinetobacter spp. in Luria-Bertani (LB) medium, filtered wastewater, and in lab- and full-scale wastewater treatment systems. Consistent with the previous reports for E. coli, results obtained with membrane hybridizations and fluorescence in situ hybridizations with Acinetobacter calcoaceticus grown in LB medium showed a more substantial and faster increase in precursor 16S rRNA levels compared to the increase in total 16S rRNA levels during exponential growth. Diluting an overnight culture of A. calcoaceticus grown in LB medium with filtered wastewater resulted in a pattern of precursor 16S rRNA levels that appeared to follow diauxic growth. In addition, fluorescence in situ hybridizations with oligonucleotide probes targeting total 16S rRNA and precursor 16S rRNA showed that individual cells of A. calcoaceticus expressed highly variable levels of precursor 16S rRNA when adapting from LB medium to filtered sewage. Precursor 16S rRNA levels of Acinetobacter spp. transiently increased when activated sludge was mixed with influent wastewater in lab- and full-scale wastewater treatment systems. These results suggest that Acinetobacter spp. experience a change in growth activity within wastewater treatment systems. PMID- 10788396 TI - Molecular ecological analysis of the succession and diversity of sulfate-reducing bacteria in the mouse gastrointestinal tract. AB - Intestinal sulfate-reducing bacteria (SRB) growth and resultant hydrogen sulfide production may damage the gastrointestinal epithelium and thereby contribute to chronic intestinal disorders. However, the ecology and phylogenetic diversity of intestinal dissimilatory SRB populations are poorly understood, and endogenous or exogenous sources of available sulfate are not well defined. The succession of intestinal SRB was therefore compared in inbred C57BL/6J mice using a PCR-based metabolic molecular ecology (MME) approach that targets a conserved region of subunit A of the adenosine-5'-phosphosulfate (APS) reductase gene. The APS reductase-based MME strategy revealed intestinal SRB in the stomach and small intestine of 1-, 4-, and 7-day-old mice and throughout the gastrointestinal tract of 14-, 21-, 30-, 60-, and 90-day-old mice. Phylogenetic analysis of APS reductase amplicons obtained from the stomach, middle small intestine, and cecum of neonatal mice revealed that Desulfotomaculum spp. may be a predominant SRB group in the neonatal mouse intestine. Dot blot hybridizations with SRB-specific 16S ribosomal DNA (rDNA) probes demonstrated SRB colonization of the cecum and colon pre- and postweaning and colonization of the stomach and small intestine of mature mice only. The 16S rDNA hybridization data further demonstrated that SRB populations were most numerous in intestinal regions harboring sulfomucin containing goblet cells, regardless of age. Reverse transcriptase PCR analysis demonstrated APS reductase mRNA expression in all intestinal segments of 30-day old mice, including the stomach. These results demonstrate for the first time widespread colonization of the mouse intestine by dissimilatory SRB and evidence of spatial-specific SRB populations and sulfomucin patterns along the gastrointestinal tract. PMID- 10788397 TI - Mutants with enhanced nitrogenase activity in hydroponic Azospirillum brasilense wheat associations. AB - The effect of a mutation affecting flocculation, differentiation into cyst-like forms, and root colonization on nitrogenase expression by Azospirillum brasilense is described. The gene flcA of strain Sp7 restored these phenotypes in spontaneous mutants of both strains Sp7 and Sp245. Employing both constitutive pLA-lacZ and nifH-lacZ reporter fusions expressed in situ, the colony morphology, colonization pattern, and potential for nitrogenase activity of spontaneous mutants and flcA Tn5-induced mutants were established. The results of this study show that the ability of Sp7 and Sp245 mutant strains to remain in a vegetative form improved their ability to express nitrogenase activity in association with wheat in a hydroponic system. Restoring the cyst formation and colonization pattern to the spontaneous mutant Sp7-S reduced nitrogenase activity rates in association with plants to that of the wild-type Sp7. Although Tn5-induced flcA mutants showed higher potentials for nitrogenase expression than Sp7, their potentials were lower than that of Sp7-S, indicating that other factors in this strain contribute to its exceptional nitrogenase activity rates on plants. The lack of lateral flagella is not one of these factors, as Sp7-PM23, a spontaneous mutant impaired in swarming and lateral-flagellum production but not in flocculation, showed wild-type nitrogenase activity and expression. The results also suggest factors of importance in evolving an effective symbiosis between Azospirillum and wheat, such as increasing the availability of microaerobic niches along the root, increased supply of carbon sources by the plant, and the retention of the bacterial cells in vegetative form for faster metabolism. PMID- 10788398 TI - Effects of glucosinolates and flavonoids on colonization of the roots of Brassica napus by Azorhizobium caulinodans ORS571. AB - Plants of Brassica napus were assessed quantitatively for their susceptibility to lateral root crack colonization by Azorhizobium caulinodans ORS571(pXLGD4) (a rhizobial strain carrying the lacZ reporter gene) and for the concentration of glucosinolates in their roots by high-pressure liquid chromatography (HPLC). High and low-glucosinolate-seed (HGS and LGS) varieties exhibited a relatively low and high percentage of colonized lateral roots, respectively. HPLC showed that roots of HGS plants contained a higher concentration of glucosinolates than roots of LGS plants. One LGS variety showing fewer colonized lateral roots than other LGS varieties contained a higher concentration of glucosinolates than other LGS plants. Inoculated HGS plants treated with the flavonoid naringenin showed significantly more colonization than untreated HGS plants. This increase was not mediated by a naringenin-induced lowering of the glucosinolate content of HGS plant roots, nor did naringenin induce bacterial resistance to glucosinolates or increase the growth of bacteria. The erucic acid content of seed did not appear to influence colonization by azorhizobia. Frequently, leaf assays are used to study glucosinolates and plant defense; this study provides data on glucosinolates and bacterial colonization in roots and describes a bacterial reporter gene assay tailored easily to the study of ecologically important phytochemicals that influence bacterial colonization. These data also form a basis for future assessments of the benefits to oilseed rape plants of interaction with plant growth-promoting bacteria, especially diazotrophic bacteria potentially able to extend the benefits of nitrogen fixation to nonlegumes. PMID- 10788399 TI - Investigation of the relationship between lysogeny and lysis of Lactococcus lactis in cheese using prophage-targeted PCR. AB - The ability of lactococcal strains to lyse (and release intracellular enzymes) during cheese manufacture can be a very desirable trait and has been associated with improvement in flavor and acceleration of cheese ripening. Using a laboratory-scale cheese manufacturing assay, the autolytic behavior of 31 strains of Lactococcus lactis was assessed. In general, marked variation was observed between strains with a 20-fold difference between the best and worst lysing strains based on the release of the intracellular enzyme lactate dehydrogenase. In a parallel experiment, the genomes of these strains were examined for the presence of prophage integrase (int) sequences by using conserved primer sequences from known lysogenic phage. Results demonstrated that the lytic behavior of lactococcal starter strains significantly correlates with the presence of prophage sequences. These results highlight not only the contribution of prophage to starter cell lysis but also the potential of PCR as a useful initial screen to assess strains for this important industrial trait. PMID- 10788400 TI - Localization of symbiotic clostridia in the mixed segment of the termite Nasutitermes takasagoensis (Shiraki). AB - Phylogeny and the distribution of symbiotic bacteria in the mixed segment of the wood-eating termite Nasutitermes takasagoensis (Shiraki) were studied. Bacterial 16S rRNA genes (rDNA) were amplified from the mixed segment of the gut by PCR, and two kinds of sequences were identified. The phylogenetic tree was constructed by neighbor-joining and maximum parsimony methods to identify symbionts harbored in the mixed segment. They are classified as low-G+C-content gram-positive bacteria and are most closely related to the genus Clostridium. The distribution of these bacteria throughout the whole gut was examined by PCR using specific primers, which suggested that they are confined to the mixed segment despite the presence of bacteria throughout the gut. In situ hybridization indicated that the symbiotic bacteria were localized to the ectoperitrophic space between the midgut wall and the peritrophic membrane in the mixed segment. Electron microscopy revealed the close association between these bacteria and the mesenteric epithelium, suggesting that they have some interactions with the gut tissue of termites. PMID- 10788401 TI - Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria. AB - A fluorescence-labeled wheat germ agglutinin staining technique (R. K. Sizemore et al., Appl. Environ. Microbiol. 56:2245-2247, 1990) was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure. PMID- 10788402 TI - Simultaneous direct counting of total and specific microbial cells in seawater, using a deep-sea microbe as target. AB - To rapidly and accurately enumerate total and specific microbes in aquatic samples, fluorescent in situ hybridization was combined with direct counting via direct immobilization of cells on a polymer-coated Nuclepore filter. The technique, named FISH-DC, achieved almost complete recovery of total cells and reproducibility of Psychrobacter pacificensis cells of deep-sea origin (error, 10 microm. Resolution limits and illumination efficiencies for different size classes of microorganisms are treated analytically. First applications for 3D tracking of protists are demonstrated. PMID- 10788410 TI - Differential damage in bacterial cells by microwave radiation on the basis of cell wall structure. AB - Microwave radiation in Escherichia coli and Bacillus subtilis cell suspensions resulted in a dramatic reduction of the viable counts as well as increases in the amounts of DNA and protein released from the cells according to the increase of the final temperature of the cell suspensions. However, no significant reduction of cell density was observed in either cell suspension. It is believed that this is due to the fact that most of the bacterial cells inactivated by microwave radiation remained unlysed. Scanning electron microscopy of the microwave-heated cells revealed severe damage on the surface of most E. coli cells, yet there was no significant change observed in the B. subtilis cells. Microwave-injured E. coli cells were easily lysed in the presence of sodium dodecyl sulfate (SDS), yet B. subtilis cells were resistant to SDS. PMID- 10788411 TI - Lack of production of electron-shuttling compounds or solubilization of Fe(III) during reduction of insoluble Fe(III) oxide by Geobacter metallireducens. AB - Studies with the dissimilatory Fe(III)-reducing microorganism Geobacter metallireducens demonstrated that the common technique of separating Fe(III) reducing microorganisms and Fe(III) oxides with semipermeable membranes in order to determine whether the Fe(III) reducers release electron-shuttling compounds and/or Fe(III) chelators is invalid. This raised doubts about the mechanisms for Fe(III) oxide reduction by this organism. However, several experimental approaches indicated that G. metallireducens does not release electron-shuttling compounds and does not significantly solubilize Fe(III) during Fe(III) oxide reduction. These results suggest that G. metallireducens directly reduces insoluble Fe(III) oxide. PMID- 10788412 TI - Identification of the pgmG gene, encoding a bifunctional protein with phosphoglucomutase and phosphomannomutase activities, in the gellan gum-producing strain Sphingomonas paucimobilis ATCC 31461. AB - The pgmG gene of Sphingomonas paucimobilis ATCC 31461, the industrial gellan gum producing strain, was cloned and sequenced. It encodes a 50,059-Da polypeptide that has phosphoglucomutase (PGM) and phosphomannomutase (PMM) activities and is 37 to 59% identical to other bifunctional proteins with PGM and PMM activities from gram-negative species, including Pseudomonas aeruginosa AlgC. Purified PgmG protein showed a marked preference for glucose-1-phosphate (G1P); the catalytic efficiency was about 50-fold higher for G1P than it was for mannose-1-phosphate (M1P). The estimated apparent K(m) values for G1P and M1P were high, 0.33 and 1.27 mM, respectively. The pgmG gene allowed the recovery of alginate biosynthetic ability in a P. aeruginosa mutant with a defective algC gene. This result indicates that PgmG protein can convert mannose-6-phosphate into M1P in the initial steps of alginate biosynthesis and, together with other results, suggests that PgmG may convert glucose-6-phosphate into G1P in the gellan pathway. PMID- 10788413 TI - Fortified sera and their use in environmental virology. AB - Four commercially available fortified sera were compared to fetal bovine serum (FBS) with regard to their ability to maintain or increase the sensitivity of the Buffalo green monkey (BGM) kidney cell line to viral infection. Nine virus strains and five wastewater samples were used. Fortified sera were comparable to FBS for the enumeration of some viruses by the plaque method and for the detection of virus in wastewater by the most-probable-number assay. PMID- 10788414 TI - Quantification of bacterial groups within human fecal flora by oligonucleotide probe hybridization. AB - To investigate the population structure of the predominant phylogenetic groups within the human adult fecal microbiota, a new oligonucleotide probe designated S G-Clept-1240-a-A-18 was designed, validated, and used with a set of five 16S rRNA targeted oligonucleotide probes. Application of the six probes to fecal samples from 27 human adults showed additivity of 70% of the total 16S rRNA detected by the bacterial domain probe. The Bacteroides group-specific probe accounted for 37% +/- 16% of the total rRNA, while the enteric group probe accounted for less than 1%. Clostridium leptum subgroup and Clostridium coccoides group-specific probes accounted for 16% +/- 7% and 14% +/- 6%, respectively, while Bifidobacterium and Lactobacillus groups made up less than 2%. PMID- 10788415 TI - Use of integrated cell culture-PCR to evaluate the effectiveness of poliovirus inactivation by chlorine. AB - Current standards, based on cell culture assay, indicate that poliovirus is inactivated by 0.5 mg of free chlorine per liter after 2 min; however, integrated cell culture-PCR detected viruses for up to 8 min of exposure to the same chlorine concentration, requiring 10 min for complete inactivation. Thus, the contact time for chlorine disinfection of poliovirus is up to five times greater than previously thought. PMID- 10788416 TI - Activities of garlic oil, garlic powder, and their diallyl constituents against Helicobacter pylori. AB - Chronic Helicobacter pylori disease is reduced with Allium vegetable intake. This study was designed to assess the in vivo anti-H. pylori potential of a variety of garlic substances. The garlic materials all showed substantial but widely differing anti-H. pylori effects against all strains and isolates tested. The MICs (range, 8 to 32 microg/ml) and minimum bactericidal concentrations (MBCs) (range, 16 to 32 microg/ml) of undiluted garlic oil (GO) were smaller than those of garlic powder (GP) (MIC range, 250 to 500 microg/ml; MBC range, 250 to 500 microg/ml) but greater than the MIC of allicin (4. 0 microg/ml) (Table 2) present in GP. Allicin (MIC, 6 microg/ml; MBC, 6 microg/ml) was more potent than diallyl disulfide (MIC range, 100 to 200 microg/ml; MBC range, 100 to 200 microg/ml), its corresponding sulfide, but of a strength similar to that of diallyl tetrasulfide (MIC range, 3 to 6 microg/ml; MBC range, 3 to 6 microg/ml). Antimicrobial activity of the diallyl sulfides increased with the number of sulfur atoms. Time course viability studies and microscopy showed dose-dependent anti-H. pylori effects with undiluted GO, GP, allicin, and diallyl trisulfide after a lag phase of ca. 1 to 2 h. Substantial in vitro anti-H. pylori effects of pure GO and GP and their diallyl sulfur components exist, suggesting their potential for in vivo clinical use against H. pylori infections. PMID- 10788417 TI - Directed transfer of large DNA fragments between Streptomyces species. AB - The biosynthesis of complex natural products in bacteria is invariably encoded within large gene clusters. Although this facilitates the cloning of such gene clusters, their heterologous expression in genetically amenable hosts remains a challenging problem, principally due to the difficulties associated with manipulating large DNA fragments. Here we describe a new method for the directed transfer of a gene cluster from one Streptomyces species to another. The method takes advantage of tra gene-mediated conjugal transfer of chromosomal DNA between actinomycetes. As proof of principle, we demonstrate transfer of the entire approximately 22-kb actinorhodin gene cluster, and also the high-frequency cotransfer of two loci that are 150 to 200 kb apart, from Streptomyces coelicolor to an engineered derivative of Streptomyces lividans. PMID- 10788418 TI - Production and characterization of monoclonal antibodies against vegetative cells of Bacillus cereus. AB - Two monoclonal antibodies (MAbs) against Bacillus cereus were produced. The MAbs (8D3 and 9B7) were selected by enzyme-linked immunosorbent assay for their reactivity with B. cereus vegetative cells. They reacted with B. cereus vegetative cells while failing to recognize B. cereus spores. Immunoblotting revealed that MAb 8D3 recognized a 22-kDa antigen, while MAb 9B7 recognized two antigens with molecular masses of approximately 58 and 62 kDa. The use of MAbs 8D3 and 9B7 in combination to develop an immunological method for the detection of B. cereus vegetative cells in foods was investigated. PMID- 10788420 TI - Nerve growth factor-induced neuronal differentiation requires generation of Rac1 regulated reactive oxygen species. AB - Nerve growth factor (NGF) stimulation of pheochromocytoma PC12 cells transiently increased the intracellular concentration of reactive oxygen species (ROS). This increase was blocked by the chemical antioxidant N-acetylcysteine and a flavoprotein inhibitor, diphenylene iodonium. NGF responses of PC12 cells, including neurite outgrowth, tyrosine phosphorylation, and AP-1 activation, was inhibited when ROS production was prevented by N-acetylcysteine and diphenylene iodonium. The expression of dominant negative Rac1N17 blocked induction of both ROS generation and morphological differentiation by NGF. The ROS produced appears to be H(2)O(2), because the introduction of catalase into the cells abolished NGF induced neurite outgrowth, ROS production, and tyrosine phosphorylation. These results suggest that the ROS, perhaps H(2)O(2), acts as an intracellular signal mediator for NGF-induced neuronal differentiation and that NGF-stimulated ROS production is regulated by Rac1 and a flavoprotein-binding protein similar to the phagocytic NADPH oxidase. PMID- 10788421 TI - A novel lipopolysaccharide response element in the Bombyx mori cecropin B promoter. AB - Cecropin B is one of the major antibacterial peptides in the silkworm, Bombyx mori. Transcription of the cecropin B gene (CecB) occurs rapidly after bacterial invasion. Using 235 base pairs (bp) of the CecB promoter region, a kappaB-related protein and two additional DNA-binding complexes (designated F2BPI and F4BP) were identified in nuclear extracts from immunized larval fat body by the electrophoretic mobility shift assay (EMSA) (1). Further EMSA analyses indicated that the F2BPI-binding site was CATTA, and that F2BPI translocated from the cytoplasm to the nucleus after infection. In a recently established B. mori cell line, NISES-BoMo-DZ, 235 bp of CecB promoter linked to a reporter luciferase was activated 6-fold by stimulation with lipopolysaccharide (LPS), which is a major trigger of CecB expression in larvae. Truncation of the F2BPI-binding site from the promoter reduced the activation 2-fold. Deletion of either of two kappaB motifs also reduced promoter activation 2-fold. Elimination of both the F2BPI binding site and the kappaB motifs resulted in the complete loss of LPS inducibility. These results indicate that the F2BPI-binding site is an LPS responsive cis-element that is necessary for full activation of CecB. PMID- 10788422 TI - Proteolytic cleavage confers nitric oxide synthase inducing activity upon prolactin. AB - Prolactin (PRL), originally associated with milk secretion, is now known to possess a wide variety of biological actions and diverse sites of production beyond the pituitary. Proteolytic cleavage is a common post-translational modification that can either activate precursor proteins or confer upon the peptide fragment unique biological actions not exerted by the parent molecule. Recent studies have demonstrated that the 16-kDa N-terminal proteolytic cleavage product of PRL (16K-PRL) acts as a potent inhibitor of angiogenesis. Despite previous demonstrations of 16K-PRL production in vivo, biological functions beyond its antiangiogenic actions remain unknown. Here we show that 16K-PRL, but not full-length PRL, acts to promote the expression of the inducible isoform of nitric oxide synthase (iNOS) and nitric oxide (*NO) production by pulmonary fibroblasts and alveolar type II cells with potency comparable with the proinflammatory cytokines interleukin-1beta, interferon gamma, and tumor necrosis factor alpha. The differential effect of 16K-PRL versus PRL occurs through a receptor distinct from known PRL receptors. Additionally, pulmonary fibroblasts express the PRL gene and endogenously produce 16K-PRL, suggesting that this pathway may serve both autocrine and paracrine roles in the regulation of *NO production. These results reveal that proteolytic cleavage of PRL confers upon this classical hormone potent iNOS inducing activity, suggesting its role in inflammatory/immune processes. PMID- 10788423 TI - Conformational changes in the cytochrome b6f complex induced by inhibitor binding. AB - Binding of stigmatellin, an inhibitor of the Q(o) site of the bc-type complexes, has been shown to induce large conformational changes of the Rieske protein in the respiratory bc(1) complex (Kim, H., Xia, D., Yu, C. A., Xia, J. Z., Kachurin, A. M., Zhang, L., Yu, L., and Deisenhofer, J. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 8026-8033; Iwata, S., Lee, J. W., Okada, K., Lee, J. K., Iwata, M., Rasmussen, B., Link, T. A., Ramaswamy, S., and Jap, B. K. (1998) Science 281, 64 71; Zhang, Z., Huang, L., Shulmeister, V. M., Chi, Y. I., Kim, K. K., Hung, L. W., Crofts, A. R., Berry, E. A., and Kim, S. H. (1998) Nature 392, 677-684). Such a movement seems necessary to shuttle electrons from the membrane-soluble quinol to the extramembrane heme of cytochrome c(1). To see whether similar changes occur in the related photosynthetic b(6)f complex, we have studied the effect of the binding of stigmatellin to the eukaryotic b(6)f complex by electron crystallography. Comparison of projection maps of thin three-dimensional crystals prepared with or without stigmatellin, and either negatively stained or embedded in glucose, reveals a similar type of movement to that observed in the bc(1) complex and suggests also the occurrence of conformational changes in the transmembrane region. PMID- 10788424 TI - Sulfite:Cytochrome c oxidoreductase from Thiobacillus novellus. Purification, characterization, and molecular biology of a heterodimeric member of the sulfite oxidase family. AB - Direct oxidation of sulfite to sulfate occurs in various photo- and chemotrophic sulfur oxidizing microorganisms as the final step in the oxidation of reduced sulfur compounds and is catalyzed by sulfite:cytochrome c oxidoreductase (EC ). Here we show that the enzyme from Thiobacillus novellus is a periplasmically located alphabeta heterodimer, consisting of a 40.6-kDa subunit containing a molybdenum cofactor and an 8.8-kDa mono-heme cytochrome c(552) subunit (midpoint redox potential, E(m8.0) = +280 mV). The organic component of the molybdenum cofactor was identified as molybdopterin contained in a 1:1 ratio to the Mo content of the enzyme. Electron paramagnetic resonance spectroscopy revealed the presence of a sulfite-inducible Mo(V) signal characteristic of sulfite:acceptor oxidoreductases. However, pH-dependent changes in the electron paramagnetic resonance signal were not detected. Kinetic studies showed that the enzyme exhibits a ping-pong mechanism involving two reactive sites. K(m) values for sulfite and cytochrome c(550) were determined to be 27 and 4 micrometer, respectively; the enzyme was found to be reversibly inhibited by sulfate and various buffer ions. The sorAB genes, which encode the enzyme, appear to form an operon, which is preceded by a putative extracytoplasmic function-type promoter and contains a hairpin loop termination structure downstream of sorB. While SorA exhibits significant similarities to known sequences of eukaryotic and bacterial sulfite:acceptor oxidoreductases, SorB does not appear to be closely related to any known c-type cytochromes. PMID- 10788425 TI - Mutations that destabilize the a' domain of human protein-disulfide isomerase indirectly affect peptide binding. AB - Protein-disulfide isomerase (PDI) is a catalyst of folding of disulfide-bonded proteins and also a multifunctional polypeptide that acts as the beta-subunit in the prolyl 4-hydroxylase alpha(2)beta(2)-tetramer (P4H) and the microsomal triglyceride transfer protein alphabeta-dimer. The principal peptide-binding site of PDI is located in the b' domain, but all domains contribute to the binding of misfolded proteins. Mutations in the C-terminal part of the a' domain have significant effects on the assembly of the P4H tetramer and other functions of PDI. In this study we have addressed the question of whether these mutations in the C-terminal part of the a' domain, which affect P4H assembly, also affect peptide binding to PDI. We observed a strong correlation between P4H assembly competence and peptide binding; mutants of PDI that failed to form a functional P4H tetramer were also inactive in peptide binding. However, there was also a correlation between inactivity in these assays and indicators of conformational disruption, such as protease sensitivity. Peptide binding activity could be restored in inactive, protease-sensitive mutants by selective proteolytic removal of the mutated a' domain. Hence we propose that structural changes in the a' domain indirectly affect peptide binding to the b' domain. PMID- 10788426 TI - Mapping the zinc ligands of S100A2 by site-directed mutagenesis. AB - S100 family proteins are characterized by short individual N and C termini and a conserved central part, harboring two Ca(2+)-binding EF-hands, one of them highly conserved among EF-hand family proteins and the other characteristic for S100 proteins. In addition to Ca(2+), several members of the S100 protein family, including S100A2, bind Zn(2+). Two regions in the amino acid sequences of S100 proteins, namely the helices of the N-terminal EF-hand motif and the very C terminal loop are believed to be involved in Zn(2+)-binding due to the presence of histidine and/or cysteine residues. Human S100A2 contains four cysteine residues, each of them located at positions that may be important for Zn(2+) binding. We have now constructed and purified 10 cysteine-deficient mutants of human S100A2 by site-directed mutagenesis and investigated the contribution of the individual cysteine residues to Zn(2+) binding. Here we show that Cys(1(3)) (the number in parentheses indicating the position in the sequence of S100A2) is the crucial determinant for Zn(2+) binding in association with conformational changes as determined by internal tyrosine fluorescence. Solid phase Zn(2+) binding assays also revealed that the C-terminal residues Cys(3(87)) and Cys(4(94)) mediated a second type of Zn(2+) binding, not associated with detectable conformational changes in the molecule. Cys(2(22)), by contrast, which is located within the first EF hand motif affected neither Ca(2+) nor Zn(2+) binding, and a Cys "null" mutant was entirely incapable of ligating Zn(2+). These results provide new information about the mechanism and the site(s) of zinc binding in S100A2. PMID- 10788427 TI - Involvement of a conserved tryptophan residue in the UDP-glucose binding of large clostridial cytotoxin glycosyltransferases. AB - Large clostridial cytotoxins catalyze the glucosylation of Rho/Ras GTPases using UDP-glucose as a cosubstrate. By site-directed mutagenesis of Clostridium sordellii lethal toxin and Clostridium difficile toxin B fragments, we identified tryptophan 102, which is located in a conserved region within the catalytic domain of all clostridial cytotoxins, to be crucial for UDP-glucose binding. Exchange of Trp-102 with alanine decreased the glucosyltransferase activity by about 1,000-fold and blocked cytotoxic activity after microinjection. Replacement of Trp-102 by tyrosine caused a 100-fold reduction in enzyme activity, indicating a partial compensation of the tryptophan function by tyrosine. Decrease in glucosyltransferase and glycohydrolase activity was caused predominantly by an increase in the K(m) for UDP-glucose of these mutants. The data indicate that the conserved tryptophan residue is implicated in the binding of the cosubstrate UDP glucose by large clostridial cytotoxins. Data bank searches revealed different groups of proteins sharing the recently identified DXD motif (Busch, C., Hofmann, F., Selzer, J., Munro, J., Jeckel, D., and Aktories, K. (1998) J. Biol. Chem. 273, 19566-19572) and a conserved region defined by a tryptophan residue equivalent to Trp-102 of C. sordellii lethal toxin. From our findings, we propose a novel family of glycosyltransferases which includes both prokaryotic and eukaryotic proteins. PMID- 10788428 TI - Interaction of UvrA and UvrB proteins with a fluorescent single-stranded DNA. Implication for slow conformational change upon interaction of UvrB with DNA. AB - UvrA and UvrB proteins play key roles in the damage recognition step in the nucleotide excision repair. However, the molecular mechanism of damage recognition by these proteins is still not well understood. In this work we analyzed the interaction between single-stranded DNA (ssDNA) labeled with a fluorophore tetramethylrhodamine (TMR) and Thermus thermophilus HB8 UvrA (ttUvrA) and UvrB (ttUvrB) proteins. TMR-labeled ssDNA (TMR-ssDNA) as well as UV irradiated ssDNA stimulated ATPase activity of ttUvrB more strongly than did normal ssDNA, indicating that this fluorescent ssDNA was recognized as damaged ssDNA. The addition of ttUvrA or ttUvrB enhanced the fluorescence intensity of TMR-ssDNA, and the intensity was much greater in the presence of ATP. Fluorescence titration indicated that ttUvrA has higher specificity for TMR-ssDNA than for normal ssDNA in the absence of ATP. The ttUvrB showed no specificity for TMR-ssDNA, but it took over 200 min for the fluorescence intensity of the ttUvrB TMR-ssDNA complex to reach saturation in the presence of ATP. This time-dependent change could be separated into two phases. The first phase was rapid, whereas the second phase was slow and dependent on ATP hydrolysis. Time dependence of ATPase activity and fluorescence polarization suggested that changes other than the binding reaction occurred during the second phase. These results strongly suggest that ttUvrB binds ssDNA quickly and that a conformational change in ttUrvB-ssDNA complex occurs slowly. We also found that DNA containing a fluorophore as a lesion is useful for directly investigating the damage recognition by UvrA and UvrB. PMID- 10788429 TI - P2Y receptor-mediated inhibition of tumor necrosis factor alpha -stimulated stress-activated protein kinase activity in EAhy926 endothelial cells. AB - In the EAhy926 endothelial cell line, UTP, ATP, and forskolin, but not UDP and epidermal growth factor, inhibited tumor necrosis factor alpha (TNFalpha)- and sorbitol stimulation of the stress-activated protein kinases, JNK, and p38 mitogen-activated protein (MAP) kinase, and MAPKAP kinase-2, the downstream target of p38 MAP kinase. In NCT2544 keratinocytes, UTP and a proteinase activated receptor-2 agonist caused similar inhibition, but in 13121N1 cells, transfected with the human P2Y(2) or P2Y(4) receptor, UTP stimulated JNK and p38 MAP kinase activities. This suggests that the effects mediated by P2Y receptors are cell-specific. The inhibitory effects of UTP were not due to induction of MAP kinase phosphatase-1, but were manifest upstream in the pathway at the level of MEK-4. The inhibitory effect of UTP was insensitive to the MEK-1 inhibitor PD 098059, changes in intracellular Ca(2+) levels, or pertussis toxin. Acute phorbol 12-myristate 13-acetate pretreatment also inhibited TNFalpha-stimulated SAP kinase activity, while chronic pretreatment reversed the effects of UTP. Furthermore, the protein kinase C inhibitors Ro318220 and Go6983 reversed the inhibitory action of UTP, but GF109203X was ineffective. These results indicate a novel mechanism of cross-talk regulation between P2Y receptors and TNFalpha stimulated SAP kinase pathways in endothelial cells, mediated by Ca(2+) independent isoforms of protein kinase C. PMID- 10788430 TI - Mechanism of activation of the chloroplast ATP synthase. A kinetic study of the thiol modulation of isolated ATPase and membrane-bound ATP synthase from spinach by Eschericia coli thioredoxin. AB - The mechanism of thiol modulation of the chloroplast ATP synthase by Escherichia coli thioredoxin was investigated in the isolated ATPase subcomplex and in the ATP synthase complex reconstituted in bacteriorhodopsin proteoliposomes. Thiol modulation was resolved kinetically by continuously monitoring ATP hydrolysis by the isolated subcomplex and ATP synthesis by proteoliposomes. The binding rate constant of reduced thioredoxin to the oxidized ATPase subcomplex devoid of its epsilon subunit could be determined. It did not depend on the catalytic turnover. Reciprocically, the catalytic turnover did not seem to depend on thioredoxin binding. Thiol modulation by Trx of the epsilon-bearing ATPase subcomplex was slow and favored the release of epsilon. The rate constant of thioredoxin binding to the membrane-bound ATP synthase increased with the protonmotive force. It was lower in the presence of ADP than in its absence, revealing a specific effect of the ATP synthase turnover on thioredoxin-gamma subunit interaction. These findings, and more especially the comparisons between the isolated ATPase subcomplex and the ATP synthase complex, can be interpreted in the frame of the rotational catalysis hypothesis. Finally, thiol modulation changed the catalytic properties of the ATP synthase, the kinetics of which became non-Michaelian. This questions the common view about the nature of changes induced by ATP synthase thiol modulation. PMID- 10788431 TI - Hgt1p, a high affinity glutathione transporter from the yeast Saccharomyces cerevisiae. AB - A high affinity glutathione transporter has been identified, cloned, and characterized from the yeast Saccharomyces cerevisiae. This transporter, Hgt1p, represents the first high affinity glutathione transporter to be described from any system so far. The strategy for the identification involved investigating candidate glutathione transporters from the yeast genome sequence project followed by genetic and physiological investigations. This approach revealed HGT1 (open reading frame YJL212c) as encoding a high affinity glutathione transporter. Yeast strains deleted in HGT1 did not show any detectable plasma membrane glutathione transport, and hgt1Delta disruptants were non-viable in a glutathione biosynthetic mutant (gsh1Delta) background. The glutathione repressible transport activity observed in wild type cells was also absent in the hgt1Delta strains. The transporter was cloned and kinetic studies indicated that Hgt1p had a high affinity for glutathione (K(m) = 54 micrometer)) and was not sensitive to competition by amino acids, dipeptides, or other tripeptides. Significant inhibition was observed, however, with oxidized glutathione and glutathione conjugates. The transporter reveals a novel class of transporters that has homologues in other yeasts and plants but with no apparent homologues in either Escherichia coli or in higher eukaryotes other than plants. PMID- 10788432 TI - Epithelial sodium channels regulate cystic fibrosis transmembrane conductance regulator chloride channels in Xenopus oocytes. AB - The cystic fibrosis transmembrane conductance regulator (CFTR), in addition to its well defined Cl(-) channel properties, regulates other ion channels. CFTR inhibits epithelial Na(+) channel (ENaC) currents in many epithelial and nonepithelial cells. Because modulation of net NaCl reabsorption has important implications in extracellular fluid volume homeostasis and airway fluid volume and composition, we investigated whether this regulation was reciprocal by examining whether ENaC regulates CFTR. Co-expression of human (h) CFTR and mouse (m) alphabetagammaENaC in Xenopus oocytes resulted in a significant, 3.7-fold increase in whole-cell hCFTR Cl(-) conductance compared with oocytes expressing hCFTR alone. The forskolin/3-isobutyl-1-methylxanthine-stimulated whole-cell conductance in hCFTR-mENaC co-injected oocytes was amiloride-insensitive, indicating an inhibition of mENaC following hCFTR activation, and it was blocked by DPC (diphenylamine-2-carboxylic acid) and was DIDS (4, 4' diisothiocyanatostilbene-2,2'-disulfonic acid)-insensitive. Enhanced hCFTR Cl(-) conductance was also observed when either the alpha- or beta-subunit of mENaC was co-expressed with hCFTR, but this was not seen when CFTR was co-expressed with the gamma-subunit of mENaC. Single Cl(-) channel analyses showed that both CFTR Cl(-) channel open probability and the number of CFTR Cl(-) channels detected per patch increased when hCFTR was co-expressed with alphabetagammamENaC. We conclude that in addition to acting as a regulator of ENaC, CFTR activity is regulated by ENaC. PMID- 10788433 TI - Divergence in regulation of nitric-oxide synthase and its cofactor tetrahydrobiopterin by tumor necrosis factor-alpha. Ceramide potentiates nitric oxide synthesis without affecting GTP cyclohydrolase I activity. AB - Synthesis of 6(R)-5,6,7,8-tetrahydrobiopterin (BH(4)), a required cofactor for inducible nitric-oxide synthase (iNOS) activity, is usually coordinately regulated with iNOS expression. In C6 glioma cells, tumor necrosis factor-alpha (TNF-alpha) concomitantly potentiated the stimulation of nitric oxide (NO) and BH(4) production induced by IFN-gamma and interleukin-1beta. Expression of both iNOS and GTP cyclohydrolase I (GTPCH), the rate-limiting enzyme in the BH(4) biosynthetic pathway, was also markedly increased, as were their activities and protein levels. Ceramide, a sphingolipid metabolite, may mediate some of the actions of TNF-alpha. Indeed, we found that bacterial sphingomyelinase, which hydrolyzes sphingomyelin and increases endogenous ceramide, or the cell permeable ceramide analogue, C(2)-ceramide, but not C(2)-dihydroceramide (N acetylsphinganine), significantly mimicked the effects of TNF-alpha on NO production and iNOS expression and activity in C6 cells. Surprisingly, although TNF-alpha increased BH(4) synthesis and GTPCH activity, neither BH(4) nor GTPCH expression was affected by C(2)-ceramide or sphingomyelinase in IFN-gamma- and interleukin-1beta-stimulated cells. It is likely that increased BH(4) levels results from increased GTPCH protein and activity in vivo rather than from reduced turnover of BH(4), because the GTPCH inhibitor, 2,4-diamino-6 hydroxypyrimidine, blocked cytokine-stimulated BH(4) accumulation. Moreover, expression of the GTPCH feedback regulatory protein, which if decreased might increase GTPCH activity, was not affected by TNF-alpha or ceramide. Treatment with the antioxidant pyrrolidine dithiocarbamate, which is known to inhibit NF kappaB and sphingomyelinase in C6 cells, or with the peptide SN-50, which blocks translocation of NF-kappaB to the nucleus, inhibited TNF-alpha-dependent iNOS mRNA expression without affecting GTPCH mRNA levels. This is the first demonstration that cytokine-stimulated iNOS and GTPCH expression, and therefore NO and BH(4) biosynthesis, may be regulated by discrete pathways. As BH(4) is also a cofactor for the aromatic amino acid hydroxylases, discovery of distinct mechanisms for regulation of BH(4) and NO has important implications for its specific functions. PMID- 10788434 TI - Hydrolysis of triple-helical collagen peptide models by matrix metalloproteinases. AB - The matrix metalloproteinase (MMP) family has been implicated in the process of a variety of diseases such as arthritis, atherosclerosis, and tumor cell metastasis. To study the mechanisms of MMP action on collagenous substrates, we have constructed homotrimeric triple-helical peptide (THP) models of the collagenase cleavage sites in types I and II collagen. The THPs incorporate either the alpha1(I)772-786 or the alpha1(II)772-783 sequence. The alpha1(I)772 786 and alpha1(II)772-783 THPs were hydrolyzed by MMP-1 at the Gly-Ile and Gly Leu bonds, respectively, analogous to the bonds cleaved in corresponding native collagens. Thus, the THPs contained all necessary information to direct MMP-1 binding and proteolysis. Subsequent investigations using the alpha1(I)772-786 THP showed hydrolysis by MMP-2, MMP-13, and a COOH-terminal domain-deleted MMP-1 (MMP 1(Delta(243-450))) but not by MMP-3 or a COOH-terminal domain-deleted MMP-3 (MMP 3(Delta(248-460))). Kinetic analyses showed a k(cat)/K(m) value of 1,808 s(-1) m( 1) for MMP-1 hydrolysis of alpha1(I)772-786 THP, approximately 10-fold lower than for type I collagen. The effect is caused primarily by relative K(m) values. MMP 2 and MMP-13 cleaved the THP more rapidly than MMP-1, but MMP-2 cleavage occurred at distinct multiple sites. Comparison of MMP-1 and MMP-1(Delta(243-450)) hydrolysis of alpha1(I)772-786 THP showed that both can cleave a triple-helical substrate with a slightly higher K(m) value for MMP-1(Delta(243-450)). We propose that the COOH-terminal domain of MMPs is necessary for orienting whole, native collagen molecules but may not be necessary for binding to and cleaving a THP. This proposal is consistent with the large distance between the MMP-1 catalytic and COOH-terminal domains observed by three-dimensional structural analysis and supports previous suggestions that the features of the catalytic domain contribute significantly toward enzyme specificity. PMID- 10788435 TI - Expression of Arabidopsis thaliana mitochondrial alanyl-tRNA synthetase is not sufficient to trigger mitochondrial import of tRNAAla in yeast. AB - It has often been suggested that precursors to mitochondrial aminoacyl-tRNA synthetases are likely carriers for mitochondrial import of tRNAs in those organisms where this process occurs. In plants, it has been shown that mutation of U(70) to C(70) in Arabidopsis thaliana tRNA(Ala)(UGC) blocks aminoacylation and also prevents import of the tRNA into mitochondria. This suggests that interaction of tRNA(Ala) with alanyl-tRNA synthetase (AlaRS) is necessary for import to occur. To test whether this interaction is sufficient to drive import, we co-expressed A. thaliana tRNA(Ala)(UGC) and the precursor to the A. thaliana mitochondrial AlaRS in Saccharomyces cerevisiae. The A. thaliana enzyme and its cognate tRNA were correctly expressed in yeast in vivo. However, although the plant AlaRS was efficiently imported into mitochondria in the transformed strains, we found no evidence for import of the A. thaliana tRNA(Ala) nor of the endogenous cytosolic tRNA(Ala) isoacceptors. We conclude that at least one other factor besides the mitochondrial AlaRS precursor must be involved in mitochondrial import of tRNA(Ala) in plants. PMID- 10788437 TI - Human chorionic gonadotropin suppresses activation of nuclear transcription factor-kappa B and activator protein-1 induced by tumor necrosis factor. AB - Human chorionic gonadotropin (hCG) suppresses cell-mediated allogeneic reactions, viral replication, tumorigenesis, and metastasis, most of which require activation of nuclear transcription factor-kappaB (NF-kappaB) and activator protein-1 (AP-1). In the present report, we investigated the effect of hCG on NF kappaB and AP-1 activated by tumor necrosis factor (TNF). Treatment of the CaCOV3 human ovarian cell line with hCG blocked TNF-induced activation of NF-kappaB, IkappaBalpha degradation, and NF-kappaB-dependent reporter gene transcription. hCG also blocked NF-kappaB activation induced by ceramide. The effect of hCG on NF-kappaB was mediated through inhibition of phosphorylation of IkappaBalpha. Because hCG also blocked TNF receptor-associated factor-2 and NF-kappaB-inducing kinase reporter gene expression, hCG must act at a step that causes phosphorylation of IkappaBalpha. AP-1 activation induced by TNF and ceramide was also suppressed by hCG. hCG abrogated the TNF-induced activation of mitogen activated protein kinase kinase and c-Jun N-terminal kinase required for NF kappaB and AP-1, respectively. Dideoxyadenosine and H-8 reversed the effect, and dibutyryl cAMP mimicked the effect, suggesting that hCG suppresses the transcription factors through cAMP-induced protein kinase A pathway. Overall, our results indicate that hCG inhibits the activation of NF-kappaB and AP-1, which may be the molecular basis by which hCG suppresses viral replication, cell proliferation, tumorigenesis, and metastasis. PMID- 10788436 TI - Protein tyrosine kinase p56lck is required for ceramide-induced but not tumor necrosis factor-induced activation of NF-kappa B, AP-1, JNK, and apoptosis. AB - Ceramide has been implicated as an intermediate in the signal transduction of several cytokines including tumor necrosis factor (TNF). Both ceramide and TNF activate a wide variety of cellular responses, including NF-kappaB, AP-1, JNK, and apoptosis. Whether ceramide transduces these signals through the same mechanism as TNF is not known. In the present study we investigated the role of the T cell-specific tyrosine kinase p56(lck) in ceramide- and TNF-mediated cellular responses by comparing the responses of Jurkat T cells with JCaM1 cells, isogeneic Lck-deficient T cells. Treatment with ceramide activated NF-kappaB, degraded IkappaBalpha, and induced NF-kappaB-dependent reporter gene expression in a time-dependent manner in Jurkat cells but not in JCaM1 cells, suggesting the critical role of p56(lck) kinase. These effects were specific to ceramide, as activation of NF-kappaB by phorbol 12-myristate 13-acetate, lipopolysaccharide, H(2)O(2), and TNF was minimally affected. p56(lck) was also found to be required for ceramide-induced but not TNF-induced AP-1 activation. Similarly, ceramide activated the protein kinases JNK and mitogen-activated protein kinase kinase in Jurkat cells but not in JCaM1 cells. Ceramide also induced cytotoxicity and activated caspases and reactive oxygen intermediates in Jurkat cells but not in JCaM1 cells. Ceramide activated p56(lck) activity in Jurkat cells. Moreover, the reconstitution of JCaM1 cells with p56(lck) tyrosine kinase reversed the ceramide induced NF-kappaB activation and cytotoxicity. Overall our results demonstrate that p56(lck) plays a critical role in the activation of NF-kappaB, AP-1, JNK, and apoptosis by ceramide but has minimal or no role in activation of these responses by TNF. PMID- 10788438 TI - Proton re-uptake partitioning between uncoupling protein and ATP synthase during benzohydroxamic acid-resistant state 3 respiration in tomato fruit mitochondria. AB - The yield of oxidative phosphorylation in isolated tomato fruit mitochondria depleted of free fatty acids remains constant when respiratory rates are decreased by a factor of 3 by the addition of n-butyl malonate. This constancy makes the determination of the contribution of the linoleic acid-induced energy dissipating pathway by the ADP/O method possible. No decrease in membrane potential is observed in state 3 respiration with increasing concentration of n butyl malonate, indicating that the rate of ATP synthesis is steeply dependent on membrane potential. Linoleic acid decreases the yield of oxidative phosphorylation in a concentration-dependent manner by a pure protonophoric process like that in the presence of FCCP. ADP/O measurements allow calculation of the part of respiration leading to ATP synthesis and the part of respiration sustained by the dissipative H(+) re-uptake induced by linoleic acid. Respiration sustained by this energy-dissipating process remains constant at a given LA concentration until more than 50% inhibition of state 3 respiration by n-butyl malonate is achieved. The energy dissipative contribution to oxygen consumption is proposed to be equal to the protonophoric activity of plant uncoupling protein divided by the intrinsic H(+)/O of the cytochrome pathway. It increases with linoleic acid concentration, taking place at the expense of ADP phosphorylation without an increase in the respiration. PMID- 10788439 TI - c-Jun and p53 activity is modulated by SUMO-1 modification. AB - The ubiquitin-related SUMO-1 molecule has been shown recently to modify covalently a number of cellular proteins including IkappaBalpha. SUMO-1 modification was found to antagonize IkappaBalpha ubiquitination and protect it from degradation. Here we identify the transcription factors c-Jun and p53, two well known targets of ubiquitin, as new substrates for SUMO-1 both in vitro and in vivo. In contrast to ubiquitin, SUMO-1 preferentially targets a single lysine residue in c-Jun (Lys-229), and the abrogation of SUMO-1 modification does not compromise its ubiquitination. Activation of Jun NH(2)-terminal kinases, which induces a reduction in c-Jun ubiquitination, similarly decreases SUMO-1 modification. Accordingly, loss of the two major Jun NH(2)-terminal kinase phosphorylation sites in c-Jun, Ser-63 and Ser-73, greatly enhances conjugation by SUMO-1. A SUMO-1- deficient c-JunK229R mutant shows an increased transactivation potential on an AP-1-containing promoter compared with wild-type c-Jun, suggesting that SUMO-1 negatively regulates c-Jun activity. As with c-Jun, SUMO-1 modification of p53 is abrogated by phosphorylation but remains unaltered upon chemical damage to DNA or Mdm2-mediated ubiquitination. The SUMO-1 attachment site in p53 (Lys-386) resides within a region known to regulate the DNA binding activity of the protein. A p53 mutant, defective for SUMO-1 conjugation, shows unaltered ubiquitination but has a slightly impaired apoptotic activity, indicating that modification by SUMO-1 might be important for the full biological activity of p53. Taken together, these data provide a first link between the SUMO-1 conjugation pathway and the regulation of transcription factors. PMID- 10788440 TI - Ceramide inhibits protein kinase B/Akt by promoting dephosphorylation of serine 473. AB - The second messenger ceramide (N-alkylsphingosine) has been implicated in a host of cellular processes including growth arrest and apoptosis. Ceramide has been reported to have effects on both protein kinases and phosphatases and may constitute an important component of stress response in various tissues. We have examined in detail the relationship between ceramide signaling and the activation of an important signaling pathway, phosphatidylinositol (PI) 3-kinase and its downstream target, protein kinase B (PKB). PKB activation was observed following stimulation of cells with the cytokine granulocyte-macrophage colony-stimulating factor. Addition of cell-permeable ceramide analogs, C(2)- or C(6)-ceramide, caused a partial loss (50-60%) of PKB activation. This reduction was not a result of decreased PI(3,4,5)P(3) or PI(3,4)P(2) generation by PI 3-kinase. Two residues of PKB (threonine 308 and serine 473) require phosphorylation for maximal PKB activation. Serine 473 phosphorylation was consistently reduced by treatment with ceramide, whereas threonine 308 phosphorylation remained unaffected. In further experiments, ceramide appeared to accelerate serine 473 dephosphorylation, suggesting the activation of a phosphatase. Consistent with this, the reduction in serine 473 phosphorylation was inhibited by the phosphatase inhibitors okadaic acid and calyculin A. Surprisingly, threonine 308 phosphorylation was abolished in cells treated with these inhibitors, revealing a novel mechanism of regulation of threonine 308 phosphorylation. These results demonstrate that PI 3-kinase dependent kinase 2-catalyzed phosphorylation of serine 473 is the principal target of a ceramide-activated phosphatase. PMID- 10788441 TI - Identification of a conserved protein that interacts with specific LIM homeodomain transcription factors. AB - Lhx3, a member of the LIM homeodomain family of transcription factors, is required for development of the pituitary and is implicated in the transcription of pituitary-specific hormone genes. In this report we describe a novel gene product, SLB, that selectively interacts with Lhx3 and the closely related LIM factor, Lhx4. The SLB cDNA encodes a 1749-residue protein that contains seven WD40 repeats near the amino terminus and a putative nuclear localization signal and does not contain other recognizable motifs. SLB is expressed in a tissue specific manner with the highest concentrations of SLB mRNA in the testis and pituitary cells. We demonstrate that SLB specifically binds to Lhx3 and Lhx4 with high affinity both in vitro and in vivo. SLB has much lower affinity or no detectable affinity for other LIM domains. An expression vector for a fragment of SLB containing the LIM-interaction domain was shown to reduce expression of Lhx3 responsive reporter genes. The ability of the LIM-interacting domain of SLB to alter reporter gene activity as well as the tissue-specific expression and the specificity of SLB binding to LIM factors suggest a possible role in modulating the transcriptional activity of specific LIM factors. PMID- 10788442 TI - Surface expression and single channel properties of KCNQ2/KCNQ3, M-type K+ channels involved in epilepsy. AB - Mutations in either KCNQ2 or KCNQ3 underlie benign familial neonatal convulsions (BFNC), an inherited epilepsy. The corresponding proteins are co-expressed in broad regions of the brain and associate to heteromeric K(+) channels. These channels mediate M-type currents that regulate neuronal excitability. We investigated the basis for the increase in currents seen after co-expressing these subunits in Xenopus oocytes. Noise analysis and single channel recordings revealed a conductance of approximately 18 pS for KCNQ2 and approximately 7 pS for KCNQ3. Different conductance levels (ranging from 8 to 22 pS) were seen upon co-expression. Their weighted average is close to that obtained by noise analysis (16 pS). The open probability of heteromeric channels was not increased significantly. Co-expression of both subunits increased the surface expression of KCNQ2 and KCNQ3 by factors of 5 and >10, respectively. A KCNQ2 mutant associated with BFNC that has a truncated cytoplasmic carboxyl terminus did not reach the surface and failed to stimulate KCNQ3 surface expression. By contrast, several BFNC-associated missense mutations in KCNQ2 or KCNQ3 did not alter their surface expression. Thus, the increase in currents seen upon co-expressing KCNQ2 and KCNQ3 is predominantly due to an increase in surface expression, which is dependent on an intact carboxyl terminus. PMID- 10788443 TI - DsbG, a protein disulfide isomerase with chaperone activity. AB - DsbG, a protein disulfide isomerase present in the periplasm of Escherichia coli, is shown to function as a molecular chaperone. Stoichiometric amounts of DsbG are sufficient to prevent the thermal aggregation of two classical chaperone substrate proteins, citrate synthase and luciferase. DsbG was also shown to interact with refolding intermediates of chemically denatured citrate synthase and prevents their aggregation in vitro. Citrate synthase reactivation experiments in the presence of DsbG suggest that DsbG binds with high affinity to early unstructured protein folding intermediates. DsbG is one of the first periplasmic proteins shown to have general chaperone activity. This ability to chaperone protein folding is likely to increase the effectiveness of DsbG as a protein disulfide isomerase. PMID- 10788444 TI - Tumor necrosis factor-alpha increases the steady-state reduction of cytochrome b of the mitochondrial respiratory chain in metabolically inhibited L929 cells. AB - The mechanism of tumor necrosis factor alpha (TNFalpha)-induced cytotoxicity in metabolically inhibited cells is unclear, although some studies have suggested that mitochondrial dysfunction and generation of reactive oxygen species may be involved. Here we studied the effect of TNFalpha on the redox state of mitochondrial cytochromes and its involvement in the generation of reactive oxygen species in metabolically inhibited L929 cells. Treatment with TNFalpha and cycloheximide (TNFalpha/CHX) induced mitochondrial cytochrome c release, increased the steady-state reduction of cytochrome b, and decreased the steady state reduction of cytochromes cc(1) and aa(3). TNFalpha/CHX treatment also induced lipid peroxidation, intracellular generation of reactive oxygen species, and cell death. Furthermore, as the cells died mitochondrial morphology changed from an orthodox to a hyperdense and condensed and finally to a swollen conformation. Antimycin A, a mitochondrial respiratory chain complex III inhibitor that binds to cytochrome b, blocked the formation of reactive oxygen species, suggesting that the free radicals are generated at the level of cytochrome b. Moreover, antimycin A, when added after 3 h of TNFalpha/CHX treatment, arrested the further release of cytochrome c and the cytotoxic response. We propose that the reduced cytochrome b promotes the formation of reactive oxygen species, lipid peroxidation of the cell membrane, and cell death. PMID- 10788445 TI - A conserved nuclear receptor consensus sequence (DR-4) mediates transcriptional activation of the chicken CYP2H1 gene by phenobarbital in a hepatoma cell line. AB - Phenobarbital-responsive DNA elements were identified in the 5'-flanking region of the chicken CYP2H1 gene by in reporter gene assays in a chicken hepatoma cell line (leghorn male hepatoma (LMH)). A 264-base pair (bp) enhancer sequence (phenobarbital-responsive unit (PBRU)) responded to phenobarbital and a variety of phenobarbital-type inducers. Analysis of putative transcription factor binding sites within the 264-bp element revealed a nuclear receptor half-site repeat (DR 4) neighboring a putative nuclear factor-1 site. This motif resembles phenobarbital response elements in the flanking regions of three phenobarbital inducible genes, rat CYP2B2, mouse Cyp2b10, and human CYP2B6. Activation of the 264-bp element was eliminated after site-directed mutagenesis of the DR-4 hexamer half-sites. Evidence for evolutionary conservation of this recognition site was indicated by activation in LMH cells of a mouse Cyp2b10 phenobarbital-responsive enhancer by the same spectrum of inducers that activate the CYP2H1 264-bp PBRU. Inhibition of this activation by okadaic acid may explain the reported inhibitory effects on induction of CYP2B1/2 and Cyp2b10 by this phosphatase inhibitor. We show that this inhibition occurs directly on the 264-bp PBRU, whereas the proximal promoter of CYP2H1 is induced by okadaic acid in reporter gene assays. These experiments exploit the unique phenobarbital inducibility of the hepatoma derived cell line LMH. PMID- 10788447 TI - Stimulation of protein kinase C modulates insulin-like growth factor-1-induced akt activation in PC12 cells. AB - Activation of protein kinase C (PKC) plays an important role in the negative regulation of receptor signaling, but its effect on insulin-like growth factor-1 (IGF-1) receptor signaling remains unclear. In this study, we characterized the intracellular pathways involved in IGF-1-induced activation of Akt and evaluated the effects of the PKC activator phorbol 12-myristate 13-acetate (PMA) on the Akt activation by IGF-1. IGF-1 induced a time- and concentration-dependent activation of Akt. The effect of IGF-1 was blocked by the phosphatidylinositide 3-kinase (PI3K) inhibitors LY294002 (50 micrometer) and wortmannin (0.5 micrometer), but not by the MEK inhibitor PD98059 (50 micrometer) or the p70 S6 kinase pathway inhibitor rapamycin (50 nm), suggesting that the stimulation of Akt by IGF-1 is mediated by the PI3K pathway. Interestingly, cotreatment with PMA (400 nm) attenuated IGF-1-induced activation of Akt. The attenuation was blocked completely by the PKC inhibitor GO6983 (0.5 micrometer), but only partially by the MEK inhibitor PD98059 (50 micrometer), indicating that MAPK-dependent and independent pathways are involved. PMA induced the activation of PKC in PC12 cells, and this induction was blocked by GO6983. These data further support the role of PKC in the effect of PMA. Moreover, PKCdelta is likely involved in the action of PMA on the basis of data obtained using isoform-specific inhibitors such as rottlerin. PMA also decreased IGF-1-induced tyrosine phosphorylation of insulin receptor substrate-1 and its association with PI3K. Taken together, these results suggest, for the first time, that stimulation of PKC modulates IGF-1 induced activation of Akt. PMID- 10788446 TI - Aminoguanidine-mediated inactivation and alteration of neuronal nitric-oxide synthase. AB - It is established that aminoguanidine (AG) is a metabolism-based inactivator of the three major isoforms of nitric-oxide synthase. AG is thought to be of potential use in diseases, such as diabetes, where pathological overproduction of NO is implicated. We show here that during the inactivation of neuronal nitric oxide synthase (nNOS) by AG that the prosthetic heme is altered, in part, to dissociable and protein-bound adducts. The protein-bound heme adduct is the result of cross-linking of the heme to residues in the oxygenase domain of nNOS. The dissociable heme product is unstable and reverts back to heme upon isolation. The alteration of the heme is concomitant with the loss in the ability to form the ferrous-CO complex of nNOS and accounts for at least two-thirds of the activity loss. Studies with [(14)C]AG indicate that alteration of the protein, in part on the reductase domain of nNOS, also occurs but at low levels. Thus, heme alteration appears to be the major cause of nNOS inactivation. The elucidation of the mechanism of inactivation of nNOS will likely lead to a better understanding of the in vivo effects of NOS inhibitors such as AG. PMID- 10788448 TI - p38JAB1 binds to the intracellular precursor of the lutropin/choriogonadotropin receptor and promotes its degradation. AB - Using the C-terminal tail of the rat lutropin/choriogonadotropin receptor (rLHR) as "bait" in a yeast two-hybrid screen resulted in the identification of p38(JAB1) (a protein initially identified as a co-activator of c-Jun) as a putative rLHR binding partner. More recently p38(JAB1) has been shown to promote the degradation of a cyclin-dependent kinase inhibitor and to be a component of the COP9 signalosome. Microscopic localization of an epitope-tagged p38(JAB1) expressed in 293 cells revealed a punctuated perinuclear and cytosolic localization, while cell fractionation studies showed that most of the p38(JAB1) was in a high speed supernatant. Co-transfection of 293 cells revealed that p38(JAB1) binds to the immature 68-kDa precursor of the rLHR that resides in the endoplasmic reticulum and promotes its degradation. It does not appear to interact with the cell surface rLHR, however, and it does not affect its expression. When transfected into HeLa cells, p38(JAB1) potentiates the transcriptional activity of c-Jun, but co-transfection with rLHR prevents this effect. We conclude that p38(JAB1) interacts with the rLHR precursor and promotes its degradation. These results reveal a novel protein binding partner of the rLHR and are consistent with current views of the functions of p38(JAB1). PMID- 10788449 TI - Functional cloning of an Arabidopsis thaliana cDNA encoding cycloeucalenol cycloisomerase. AB - Plants and certain protists use cycloeucalenol cycloisomerase (EC ) to convert pentacyclic cyclopropyl sterols to conventional tetracyclic sterols. We used a novel complementation strategy to clone a cycloeucalenol cycloisomerase cDNA. Expressing an Arabidopsis thaliana cycloartenol synthase cDNA in a yeast lanosterol synthase mutant provided a sterol auxotroph that could be genetically complemented with the isomerase. We transformed this yeast strain with an Arabidopsis yeast expression library and selected sterol prototrophs to obtain a strain that accumulated biosynthetic ergosterol. The novel phenotype was conferred by an Arabidopsis cDNA that potentially encodes a 36-kDa protein. We expressed this cDNA (CPI1) in Escherichia coli and showed by gas chromatography mass spectrometry that extracts from this strain isomerized cycloeucalenol to obtusifoliol in vitro. The cDNA will be useful for obtaining heterologously expressed protein for catalytic studies and elucidating the in vivo roles of cyclopropyl sterols. PMID- 10788450 TI - Transcriptional regulation of glutaredoxin and thioredoxin pathways and related enzymes in response to oxidative stress. AB - We examined the in vivo expression of up to 16 genes encoding for components of both glutaredoxin and thioredoxin systems and for members of the OxyR and SoxRS regulons. We demonstrated that grxA (Grx1) transcription is triggered in bacteria lacking Trx1 (trxA) and GSH (gshA) in an OxyR-dependent manner. We also indicated that, unlike OxyR, SoxR is not constitutively activated in the oxidizing environment of trxA gshA mutants. We discovered that the lack of Trx1 plus GSH increases the steady-state levels of Trx reductase (trxB) and Trx2 (trxC) transcripts. This increase and the trxB and trxC up-regulation caused by the constitutive oxyR2 allele indicate that OxyR also plays a role in the regulation of the thioredoxin pathway. On the contrary, no change in the expression of genes for Trx1, Grx2, and Grx3 was observed. Transcription of nrdAB (RRase) was not induced by oxidative stress yet was induced by hydroxyurea (RRase inhibitor). Induction level was as the enhanced nrdAB basal expression of trxA grxA mutants, indicating that RRase operation without Trx1 and Grx1 must lead to disturbances sensed as those caused by hydroxyurea. We also demonstrated an inverse relation between nrdAB expression and that of genes coding for components of both glutaredoxin (grxA, gorA) and thioredoxin (trxB, trxC) systems. PMID- 10788451 TI - Ca2+ depletion and inositol 1,4,5-trisphosphate-evoked activation of Ca2+ entry in single guinea pig hepatocytes. AB - Store-operated Ca(2+) entry was investigated by monitoring the Ca(2+)-dependent K(+) permeability in voltage-clamped guinea pig hepatocytes. In physiological conditions, intracellular Ca(2+) stores are discharged following agonist stimulation, but depletion of this stores can be achieved using Ca(2+)-Mg(2+) ATPase inhibitors such as 2,5-di(tert-butyl)-1,4-benzohydroquinone and thapsigargin. The effect of internal Ca(2+) store depletion on Ca(2+) influx was tested in single cells using inositol 1,4,5-trisphosphate (InsP(3)) release from caged InsP(3) after treatment of the cells with 2, 5-di(tert-butyl)-1,4 benzohydroquinone or thapsigargin in Ca(2+)-free solutions. We show that the photolytic release of 1-d-myo-inositol 1,4-bisphosphate 5-phosphorothioate, a stable analog of InsP(3), and Ca(2+) store depletion have additive effects to activate a high level of Ca(2+) entry in single guinea pig hepatocytes. These results suggest that there is a direct functional interaction between InsP(3) receptors and Ca(2+) channels in the plasma membrane, although the nature of these Ca(2+) channels in hepatocytes is unclear. PMID- 10788452 TI - Na+ binding of V-type Na+-ATPase in Enterococcus hirae. AB - Rotation catalysis theory has been successfully applied to the molecular mechanism of the ATP synthase (F(0)F(1)-ATPase) and probably of the vacuolar ATPase. We investigated the ion binding step to Enterococcus hirae Na(+) translocating V-ATPase. The kinetics of Na(+) binding to purified V-ATPase suggested 6 +/- 1 Na(+) bound/enzyme molecule, with a single high affinity (K(d(Na(+()))) = 15 +/- 5 micrometer). The number of cation binding sites is consistent with the model that V-ATPase proteolipids form a rotor ring consisting of hexamers, each having one cation binding site. Release of the bound (22)Na(+) from purified molecules in a chasing experiment showed two phases: a fast component (about two-thirds of the total amount of bound Na(+); k(exchange) > 1.7 min(-1)) and a slow component (about one-third of the total; k(exchange) = 0.16 min(-1)), which changes to the fast component by adding ATP or ATPgammaS. This suggested that about two-thirds of the Na(+) binding sites of the Na(+)-ATPase are readily accessible from the aqueous phase and that the slow component is important for the transport reaction. PMID- 10788453 TI - Inhibition of AT1 receptor internalization by concanavalin A blocks angiotensin II-induced ERK activation in vascular smooth muscle cells. Involvement of epidermal growth factor receptor proteolysis but not AT1 receptor internalization. AB - Recent studies of beta(2)-adrenergic receptor suggest that agonist-promoted receptor internalization may play an important role in extracellular signal regulated kinase (ERK) activation by G protein-coupled receptors. In the present study, we explored the effects of angiotensin II (Ang II) type-1 receptor (AT(1)) internalization on Ang II-induced activation of ERK using the receptor internalization blocker concanavalin A (ConA) and the carboxyl terminus-truncated receptor mutants with impaired internalization. ConA inhibited AT(1) receptor internalization without affecting ligand binding to the receptor, Ang II-induced generation of second messengers, and activation of tyrosine kinases Src and Pyk2 in vascular smooth muscle cells (VSMC). ConA blocked ERK activation evoked by Ang II and the calcium ionophore A23187. Impairment of AT(1) receptor internalization by truncating the receptor carboxyl terminus did not affect Ang II-induced ERK activation. ConA induced proteolytic cleavage of the epidermal growth factor (EGF) receptor at carboxyl terminus and abolished Ang II-induced transactivation of the EGF receptor, which is critical for ERK activation by Ang II in VSMC. ConA also induced proteolysis of erbB-2 but not platelet-derived growth factor receptor. Thus, ConA blocks Ang II-induced ERK activation in VSMC through a distinct mechanism, the ConA-mediated proteolysis of the EGF receptor. PMID- 10788454 TI - Regulation of prostaglandin biosynthesis by nitric oxide is revealed by targeted deletion of inducible nitric-oxide synthase. AB - We investigated the effects of targeted deletion of the inducible NO synthase (iNOS) gene on the formation of prostaglandins in vivo and ex vivo. Peritoneal macrophages were obtained from control and iNOS-deficient mice, and prostaglandin E(2) (PGE(2)) was quantified after stimulation with gamma-interferon and lipopolysaccharide to induce COX-2. Total nitrate and nitrite production was completely abolished in cells from iNOS-deficient animals compared with control cells. PGE(2) formation by cells from iNOS-deficient animals was decreased compared with cells from control animals 80% at 12 h (0.85 +/- 0.90 ng/10(6) cells versus 15.4 +/- 2.1 ng/10(6) cells, p < 0.01) and 74% at 24 h (9.4 +/- 4.3 ng/10(6) cells versus 36.8 +/- 4.1 ng/10(6) cells, p < 0.01). COX-2 protein expression was not significantly different in cells from control or knockout animals. Levels of PGE(2) in the urine of iNOS-deficient mice were decreased 78% (0.24 +/- 0.14 ng/mg of creatinine versus 1.09 +/- 0.66 ng/mg of creatinine, p < 0.01) compared with control animals. In addition, the levels of urinary F(2) isoprostanes, an index of endogenous oxidant stress, were significantly decreased in iNOS-deficient animals. In contrast, the levels of thromboxane B(2) derived from platelets allowed to aggregate ex vivo were significantly increased in iNOS deficient mice compared with wild-type mice. These studies support the hypothesis that NO and/or NO-derived species modulate cyclooxygenase activity and eicosanoid production in vivo. PMID- 10788455 TI - Evidence for long range allosteric interactions between the extracellular and cytoplasmic parts of bacteriorhodopsin from the mutant R82A and its second site revertant R82A/G231C. AB - Evidence is presented for long range interactions between the extracellular and cytoplasmic parts of the heptahelical membrane protein bacteriorhodopsin in the mutant R82A and its second site revertant R82A/G231C. (i) In the double mutants R82A/G72C and R82A/A160C, with the cysteine mutation on the extracellular or cytoplasmic surface, respectively, the photocycle is the same as in the single mutant R82A with an accelerated deprotonation of the Schiff base and a reversed order of proton release and uptake. Proton release and uptake kinetics were measured directly at either surface by using the unique cysteine residue as attachment site for the pH indicator fluorescein. Whereas in wild type proton uptake on the cytoplasmic surface occurs during the M-decay (tau approximately 8 ms), in R82A it occurs already during the first phase of the M-rise (tau < 1 microseconds). (ii) The introduction of a second mutation at the cytoplasmic surface in position 231 (helix G) restores wild type ground state absorption properties, kinetics of photocycle and of proton release, and uptake in the mutant R82A/G231C. In addition, kinetic H/D isotope effects provide evidence that the proton release mechanism in R82A/G231C and in wild type is similar. These results suggest the existence of long range interactions between the cytoplasmic and extracellular surface domains of bacteriorhodopsin mediated by salt bridges and hydrogen-bonded networks between helices C (Arg-82) and G (Asp-212 and Gly 231). Such long range interactions are expected to be of functional significance for activation and signal transduction in heptahelical G-protein-coupled receptors. PMID- 10788456 TI - Zn2+ inhibits alpha-ketoglutarate-stimulated mitochondrial respiration and the isolated alpha-ketoglutarate dehydrogenase complex. AB - Intracellular free Zn(2+) is elevated in a variety of pathological conditions, including ischemia-reperfusion injury and Alzheimer's disease. Impairment of mitochondrial respiration is also associated with these pathological conditions. To test whether elevated Zn(2+) and impaired respiration might be linked, respiration of isolated rat liver mitochondria was measured after addition of Zn(2+). Zn(2+) inhibition (K(i)(app) = approximately 1 micrometer) was observed for respiration stimulated by alpha-ketoglutarate at concentrations well within the range of intracellular Zn(2+) reported for cultured hepatocytes. The bc(1) complex is inhibited by Zn(2+) (Link, T. A., and von Jagow, G. (1995) J. Biol. Chem. 270, 25001-25006). However, respiration stimulated by succinate (K(i)(app) = approximately 6 micrometer) was less sensitive to Zn(2+), indicating the existence of a mitochondrial target for Zn(2+) upstream from bc(1) complex. Purified pig heart alpha-ketoglutarate dehydrogenase complex was strongly inhibited by Zn(2+) (K(i)(app) = 0.37 +/- 0.05 micrometer). Glutamate dehydrogenase was more resistant (K(i)(app) = 6 micrometer), malate dehydrogenase was unaffected, and succinate dehydrogenase was stimulated by Zn(2+). Zn(2+) inhibition of alpha-ketoglutarate dehydrogenase complex required enzyme cycling and was reversed by EDTA. Reversibility was inversely related to the duration of exposure and the concentration of Zn(2+). Physiological free Zn(2+) may modulate hepatic mitochondrial respiration by reversible inhibition of the alpha ketoglutarate dehydrogenase complex. In contrast, extreme or chronic elevation of intracellular Zn(2+) could contribute to persistent reductions in mitochondrial respiration that have been observed in Zn(2+)-rich diseased tissues. PMID- 10788457 TI - Binding of the cGMP-gated channel to the Na/Ca-K exchanger in rod photoreceptors. AB - The intracellular Ca(2+) concentration in rod outer segments of vertebrate photoreceptors is controlled by Ca(2+) influx through cGMP-gated channels and by Ca(2+) efflux driven by Na/Ca-K exchangers. Previously, we suggested that channel and exchanger are associated (Bauer, P. J., and Drechsler, M. (1992) J. Physiol. (Lond. ) 451, 109-131). This suggestion has been thoroughly examined using a variety of biochemical approaches. First, we took advantage of the fact that cGMP gated channels bind calmodulin (CaM). Using CaM affinity chromatographic purification of the channel in 10 mm CHAPS, a significant fraction of exchanger was co-eluted with the channel indicating a binding affinity between channel and exchanger. Binding of channel and exchanger was examined more directly by cross linking of proteins in the rod outer segment membranes. Activation of the channel with cyclic 8-bromo-GMP lead to exposure of a cysteine, which allowed cross linking of the channel to the exchanger with the thiol-specific reagent dl-1,4 bismaleimido-2,3-butanediol. Cleavage of the cross-links and electrophoretic analysis indicated that a cross-link between the alpha-subunit of the channel and the exchanger formed. Furthermore, a cross-link between two adjacent alpha subunits of the channel was found, suggesting that the alpha-subunits of the native channel are dimerized. Further support for an interaction between alpha subunit and exchanger was obtained by in vitro experiments. Specific binding of the exchanger to the alpha-subunit but not to the beta-subunit of the channel was observed in Western blots of purified channel incubated with purified exchanger. This study suggests that two exchanger molecules bind to one cGMP-gated channel and, more specifically, that binding of exchanger molecules occurs at the alpha subunits, which in the native channel are dimerized. The implications of these findings regarding the possibility of local Ca(2+) signaling in vertebrate photoreceptors will be discussed. PMID- 10788458 TI - Monitoring enzyme catalysis with mass spectrometry. AB - Mass spectrometry is a rapid, sensitive, and accurate quantitative approach for the direct monitoring of enzyme-catalyzed reactions that does not require a chromophore or radiolabeling and thus provides a viable alternative to existing analytical techniques. In this study the proteolysis of intact viral capsid proteins, the alpha-glucosidase-catalyzed hydrolysis of p-nitrophenyl-alpha glucopyranoside and the lipoprotein lipase-catalyzed ester hydrolysis of resorufin were examined. Matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry were used to examine the proteolysis of viral protein capsids, providing information about capsid dynamics and the stabilizing force of viral protein/RNA interactions. In addition, k(cat) and K(m) values of enzyme-catalyzed hydrolysis were obtained (without the use of a chromophore). These results also demonstrate the effect an unnatural substrate can have on enzyme activity. Overall, mass spectrometry provides for efficient and quantitative analysis of enzyme-catalyzed reactions, as well as the direct observation of reaction dynamics. PMID- 10788459 TI - Mutational analysis of the substrate binding/catalytic domains of human M form and P form phenol sulfotransferases. AB - Human monoamine (M) form and simple phenol (P) form phenol sulfotransferases (PSTs) are greater than 93% identical in their primary sequences and yet display distinct substrate specificities and other enzymatic properties. Through the generation and characterization of a series of chimeric PSTs, we have previously demonstrated two highly variable regions within their sequences to be responsible for determining their substrate phenotypes (Sakakibara, Y., Takami, Y., Nakayama, T., Suiko, M., and Liu, M.-C. (1998) J. Biol. Chem. 273, 6242-6247). By employing the site-directed mutagenesis technique, the present study aims to identify and quantitatively evaluate the specific amino acid residues critical to the substrate binding and catalysis in these two enzymes. Twelve mutated M-PSTs and seven mutated P-PSTs were generated, expressed, and purified. Enzymatic characterization showed that, of the twelve mutated M-PSTs, mutations at residues Asp-86, Glu-89, and Glu-146 resulted in a dramatic decrease in V(max)/K(m) with dopamine as substrate, being greater than 450 times for the D86A/E89I/E146A mutated M-PST. With p-nitrophenol as substrate, the V(max)/K(m) determined for the D86A/E89I/E146A-mutated M-PST increased more than 25 times and approached that determined for the wild-type P-PST. These results indicated that the concerted action of the three mutated residues (D86A, E89I, and E146A) is sufficient for the conversion of the substrate phenotype of M-PST to that of P PST. Among the mutated P-PSTs, the I89E- and A146E-mutated P-PSTs displayed considerable deviations in V(max)/K(m) with dopamine or p-nitrophenol as substrate. No corresponding changes, however, were detected with the opposite compound as substrate. These results indicated that, in contrast to M-PST, mutations at Ala-86, Ile-89, and Ala-146 to the corresponding residues in M-PST are not sufficient for rendering the change of P-PST substrate phenotype to that of M-PST. For both M-PSTs and P-PSTs, mutations at Lys-48 or His-108 led to the loss of sulfotransferase activities, indicating their importance in the catalytic mechanism. PMID- 10788460 TI - Effectors increase the affinity of ADP-ribosylation factor for GTP to increase binding. AB - The stoichiometry of the binding of GTP to ADP-ribosylation factor (ARF) proteins, normally quite low at approximately 0.05 mol/mol protein, was found to increase to a maximum of 1 mol/mol in the presence of effectors. The mechanism of this action was found to result from the ability of these effectors to increase the affinity of ARF for activating guanine nucleotide triphosphates. The existence of a conformation of ARF with low affinity (>100 micrometer) for GTP is proposed. The actions of effectors to increase the equilibrium binding of GTP is interpreted as evidence that these same effectors interact with and modulate the affinity of the inactive ARF for GTP. A new model for these interactions among ARF, effectors, and GTP is proposed, and a preliminary test in cells is supportive of these observations with relevance to signaling in cells. PMID- 10788461 TI - Munc18-2, a functional partner of syntaxin 3, controls apical membrane trafficking in epithelial cells. AB - The Sec1-related proteins bind to syntaxin family t-SNAREs with high affinity, thus controlling the interaction of syntaxins with their cognate SNARE partners. Munc18-2 is a Sec1 homologue enriched in epithelial cells and forms a complex with syntaxin 3, a t-SNARE localized to the apical plasma membrane. We generated here a set of Munc18-2 point mutants with substitutions in conserved amino acid residues. The mutants displayed a spectrum of different syntaxin binding efficiencies. The in vitro and in vivo binding patterns were highly similar, and the association of the Munc18-2 variants with syntaxin 3 correlated well with their ability to displace SNAP-23 from syntaxin 3 complexes when overexpressed in Caco-2 cells. Even the Munc18-2 mutants that do not detectably bind syntaxin 3 were membrane associated in Caco-2 cells, suggesting that the syntaxin interaction is not the sole determinant of Sec1 protein membrane attachment. Overexpression of the wild-type Munc18-2 was shown to inhibit the apical delivery of influenza virus hemagglutinin (HA). Interestingly, mutants unable to bind syntaxin 3 behaved differently in the HA transport assay. While one of the mutants tested had no effect, one inhibited and one enhanced the apical transport of HA. This implies that Munc18-2 function in apical membrane trafficking involves aspects independent of the syntaxin 3 interaction. PMID- 10788462 TI - Anandamide uptake by human endothelial cells and its regulation by nitric oxide. AB - Anandamide (AEA) has vasodilator activity, which can be terminated by cellular re uptake and degradation. Here we investigated the presence and regulation of the AEA transporter in human umbelical vein endothelial cells (HUVECs). HUVECs take up AEA by facilitated transport (apparent K(m) = 190 +/- 10 nm and V(max) = 45 +/ 3 pmol. min(-1).mg(-1) protein), which is inhibited by alpha-linolenoyl-vanillyl amide and N-(4-hydroxyphenyl)-arachidonoylamide, and stimulated up to 2.2-fold by nitric oxide (NO) donors. The NO scavenger hydroxocobalamin abolishes the latter effect, which is instead enhanced by superoxide anions but inhibited by superoxide dismutase and N-acetylcysteine, a precursor of glutathione synthesis. Peroxynitrite (ONOO(-)) causes a 4-fold activation of AEA transport into cells. The HUVEC AEA transporter contributes to the termination of a typical type 1 cannabinoid receptor (CB(1)) -mediated action of AEA, i.e. the inhibition of forskolin-stimulated adenylyl cyclase, because NO/ONOO(-) donors and alpha linolenoyl-vanillyl-amide/N-(4-hydroxyphenyl)-arachidonoylamide were found to attenuate and enhance, respectively, this effect of AEA. Consistently, activation of CB(1) cannabinoid receptors by either AEA or the cannabinoid HU-210 caused a stimulation of HUVEC inducible NO synthase activity and expression up to 2.9- and 2. 6-fold, respectively. Also these effects are regulated by the AEA transporter. HU-210 enhanced AEA uptake by HUVECs in a fashion sensitive to the NO synthase inhibitor Nomega-nitro-l-arginine methyl ester. These findings suggest a NO mediated regulatory loop between CB(1) cannabinoid receptors and AEA transporter. PMID- 10788463 TI - Estrogen receptor-KRAB chimeras are potent ligand-dependent repressors of estrogen-regulated gene expression. AB - As an approach to targeted repression of genes of interest, we describe the development of human estrogen receptor (ER) alpha-KRAB repressor domain chimeras that are potent ligand-dependent repressors of the transcription of estrogen response element (ERE)-containing promoters and analyze their mechanisms of action. Repression by the KRAB domain was dominant over transactivation mediated by ER AF1 and AF2. An ERE and an ER ligand (estrogen or antiestrogen) were required for repression. Studies with several promoters and cell lines demonstrated that the presence of EREs, rather than the capacity for estrogen induction, determines the potential for repression of a gene by the KRAB-ERalpha KRAB (KERK) chimera. A single consensus ERE was sufficient for repression, but the KERK chimera was unable to suppress transcription from the imperfect ERE in the native pS2 promoter. We recently reported mutations that enhance binding of a steroid receptor DNA-binding domain to the ERE. Introducing these mutations into wild-type ER enhanced transactivation from the pS2 ERE. Insertion of these mutations into KERK created the novel repressor KERK-3M, which is a potent repressor of both ER-induced and basal transcription on a promoter containing the pS2 ERE. These modified ER-KRAB chimeras should prove useful as new tools for the functional analysis and repression of ER-regulated genes. PMID- 10788464 TI - Thrombin responses in human endothelial cells. Contributions from receptors other than PAR1 include the transactivation of PAR2 by thrombin-cleaved PAR1. AB - The recent identification of two new thrombin receptors, PAR3 and PAR4, led us to re-examine the basis for endothelial cell responses to thrombin. Human umbilical vein endothelial cells (HUVEC) are known to express PAR1 and the trypsin/tryptase receptor, PAR2. Northern blots detected both of those receptors and, to a lesser extent, PAR3, but PAR4 message was undetectable and there was no response to PAR4 agonist peptides. To determine whether PAR3 or any other receptor contributes to thrombin signaling in HUVEC, PAR1 cleavage was blocked with two selective antibodies and PAR1 activation was inhibited with the antagonist, BMS200261. The antibodies completely inhibited HUVEC responses to thrombin, but BMS200261 was only partly effective, even though separate studies established that the antagonist completely inhibits PAR1 signaling at the concentrations used. Since peptides mimicking the PAR1 tethered ligand domain can also activate PAR2, we asked whether the remaining thrombin response in the presence of the antagonist could be due in part to the intermolecular transactivation of PAR2 by cleaved PAR1. Evidence that transactivation can occur was obtained in COS-7 cells co expressing PAR2 and a variant of PAR1 that can be cleaved, but not signal. There was a substantial response to thrombin only in cells expressing both receptors. Conversely, in HUVEC, complete blockade of the thrombin response by the PAR1 antagonist occurred only when signaling through PAR2 was also blocked. From these observations we conclude that 1) PAR1 is the predominant thrombin receptor expressed in HUVEC and cleavage of PAR1 is required for endothelial cell responses to thrombin; 2) although PAR3 may be expressed, there is still no evidence that it mediates thrombin responses; 3) PAR4 is not expressed on HUVEC; and 4) transactivation of PAR2 by cleaved PAR1 can contribute to endothelial cell responses to thrombin, particularly when signaling through PAR1 is blocked. Such transactivation may limit the effectiveness of PAR1 antagonists, which compete with the tethered ligand domain rather than preventing PAR1 cleavage. PMID- 10788465 TI - Isolation and characterization of peroxisome proliferator-activated receptor (PPAR) interacting protein (PRIP) as a coactivator for PPAR. AB - We previously isolated and identified steroid receptor coactivator-1 (SRC-1) and peroxisome proliferator-activated receptor (PPAR)-binding protein (PBP/PPARBP) as coactivators for PPAR, using the ligand-binding domain of PPARgamma as bait in a yeast two-hybrid screening. As part of our continuing effort to identify cofactors that influence the transcriptional activity of PPARs, we now report the isolation of a novel coactivator from mouse, designated PRIP (peroxisome proliferator-activated receptor interacting protein), a nuclear protein with 2068 amino acids and encoded by 13 exons. Northern analysis showed that PRIP mRNA is ubiquitously expressed in many tissues of adult mice. PRIP contains two LXXLL signature motifs. The amino-terminal LXXLL motif (amino acid position 892 to 896) of PRIP was found to be necessary for nuclear receptor interaction, but the second LXXLL motif (amino acid position 1496 to 1500) appeared unable to bind PPARgamma. Deletion of the last 12 amino acids from the carboxyl terminus of PPARgamma resulted in the abolition of the interaction between PRIP and PPARgamma. PRIP also binds to PPARalpha, RARalpha, RXRalpha, ER, and TRbeta1, and this binding is increased in the presence of specific ligands. PRIP acts as a strong coactivator for PPARgamma in the yeast and also potentiates the transcriptional activities of PPARgamma and RXRalpha in mammalian cells. A truncated form of PRIP (amino acids 786-1132) acts as a dominant-negative repressor, suggesting that PRIP is a genuine coactivator. PMID- 10788466 TI - Lamprey hemoglobin. Structural basis of the bohr effect. AB - Lampreys, among the most primitive living vertebrates, have hemoglobins (Hbs) with self-association and ligand-binding properties very different from those that characterize the alpha(2)beta(2) tetrameric Hbs of higher vertebrates. Monomeric, ligated lamprey Hb self-associates to dimers and tetramers upon deoxygenation. Dissociation to monomers upon oxygenation accounts for the cooperative binding of O(2) and its pH dependence. Honzatko and Hendrickson (Honzatko, R. B., and Hendrickson, W. A. (1986) Proc. Natl. Acad. Sci. U. S. A 83, 8487-8491) proposed that the dimeric interface of the Hb resembles either the alpha(1)beta(2) interface of mammalian Hbs or the contacts in clam Hb where the E and F helices form the interface. Perutz (Perutz, M. F. (1989) Quart. Rev. Biophys. 2, 139- 236) proposed a version of the clam model in which the distal histidine swings out of the heme pocket upon deoxygenation to form a bond with a carboxyl group of a second monomer. The sedimentation behavior and oxygen equilibria of nine mutants of the major Hb component, PMII, from Petromyzon marinus have been measured to test these models. The results strongly support a critical role of the E helix and the AB corner in forming the subunit interface in the dimer and rule out the alpha(1)beta(2) model. The pH dependence of both the sedimentation equilibrium and the oxygen binding of the mutant E75Q indicate that Glu(75) is one of two groups responsible for the Bohr effect. Changing the distal histidine 73 to glutamine almost completely abolishes the self-association of the deoxy-Hb and causes a large increase in O(2) affinity. The recent x-ray crystallographic determination of the structure of deoxy lamprey Hb, reported after the completion of this work (Heaslet, H. A., and Royer, W. E. (1999) Structure 7, 517-526), shows that the dimer interface does involve the E helix and the AB corner, supporting the measurements and interpretations reported here. PMID- 10788467 TI - Role of individual monomers of a dimeric initiator protein in the initiation and termination of plasmid rolling circle replication. AB - Plasmids of the pT181 family encode initiator proteins that act as dimers during plasmid rolling circle (RC) replication. These initiator proteins bind to the origin of replication through a sequence-specific interaction and generate a nick at the origin that acts as the primer for RC replication. Previous studies have demonstrated that the initiator proteins contain separate DNA binding and nicking closing domains, both of which are required for plasmid replication. The tyrosine residue at position 191 of the initiator RepC protein of pT181 is known to be involved in nicking at the origin. We have generated heterodimers of RepC that consist of different combinations of wild type, DNA binding, and nicking mutant monomers to identify the role of each of the two monomers in RC replication. One monomer with DNA binding activity was sufficient for the targeting of the initiator to the origin, and the presence of Tyr-191 in one monomer was sufficient for the initiation of replication. On the other hand, a dimer consisting of one monomer defective in DNA binding and the other defective in origin nicking failed to initiate replication. Our results demonstrate that the monomer that promotes sequence-specific binding to the origin must also nick the DNA to initiate replication. Interestingly, whereas Tyr-191 of the initiator was required for nicking at the origin to initiate replication, it was dispensable for termination, suggesting that alternate amino acids in the initiator may promote termination but not initiation. PMID- 10788468 TI - Evidence for a concerted mechanism of ubiquinol oxidation by the cytochrome bc1 complex. AB - To better understand the mechanism of divergent electron transfer from ubiquinol to the iron-sulfur protein and cytochrome b(L) within the cytochrome bc(1) complex, we have examined the effects of antimycin on the presteady state reduction kinetics of the bc(1) complex in the presence or absence of endogenous ubiquinone. When ubiquinone is present, antimycin slows the rate of cytochrome c(1) reduction by approximately 10-fold but had no effect upon the rate of cytochrome c(1) reduction in bc(1) complex lacking endogenous ubiquinone. In the absence of endogenous ubiquinone cytochrome c(1), reduction was slower than when ubiquinone was present and was similar to that in the presence of ubiquinone plus antimycin. These results indicate that the low potential redox components, cytochrome b(H) and b(L), exert negative control on the rate of reduction of cytochrome c(1) and the Rieske iron-sulfur protein at center P. If electrons cannot equilibrate from cytochrome b(H) and b(L) to ubiquinone, partial reduction of the low potential components slows reduction of the high potential components. We also examined the effects of decreasing the midpoint potential of the iron sulfur protein on the rates of cytochrome b reduction. As the midpoint potential decreased, there was a parallel decrease in the rate of b reduction, demonstrating that the rate of b reduction is dependent upon the rate of ubiquinol oxidation by the iron-sulfur protein. Together these results indicate that ubiquinol oxidation is a concerted reaction in which both the low potential and high potential redox components control ubiquinol oxidation at center P, consistent with the protonmotive Q cycle mechanism. PMID- 10788470 TI - Design and analysis of an engineered human interleukin-10 monomer. AB - A monomeric form of human interleukin 10 (IL-10M1) has been engineered for detailed structure-function studies on IL-10 and its receptor complexes. Wild type IL-10 (wtIL-10) is a domain swapped dimer whose structural integrity depends on the intertwining of two peptide chains. wtIL-10 was converted to a monomeric isomer by inserting 6 amino acids into the loop connecting the swapped secondary structural elements. Characterization of IL-10M1 by mass spectroscopy, size exclusion chromatography, cross-linking, and circular dichroism shows that IL 10M1 is a stable alpha-helical monomer at physiological pH whose three dimensional structure closely resembles one domain of wtIL-10. As previously reported, incubation of wtIL-10 with a soluble form of the IL-10Ralpha (sIL 10Ralpha) generates a complex that consists of 2 wtIL-10 molecules and 4 sIL 10Ralphas. In contrast, IL-10M1 forms a 1:1 complex with the sIL-10Ralpha. Characterization of the interaction using isothermal titration calorimetry confirmed the 1:1 stoichiometry and yielded a dissociation constant of 30 nm with an apparent binding enthalpy of -12.2 kcal/mol. Despite forming a 1:1 complex, IL 10M1 is biologically active in cellular proliferation assays. These results indicate that the 1:1 interaction between IL-10M1 and IL-10Ralpha is sufficient for recruiting the signal transducing receptor chain (IL-10Rbeta) into the signaling complex and eliciting IL-10 cellular responses. PMID- 10788469 TI - High-resolution NMR spectroscopy of lipid A molecules containing 4-amino-4-deoxy L-arabinose and phosphoethanolamine substituents. Different attachment sites on lipid A molecules from NH4VO3-treated Escherichia coli versus kdsA mutants of Salmonella typhimurium. AB - When Escherichia coli are grown on LB broth containing 25 mm NH(4)VO(3), complex modifications to the lipid A anchor of lipopolysaccharide are induced. Six modified lipid As (EV1-EV6) have been purified. Many of these variants possess 4 amino-4-deoxy-l-arabinose (l-Ara4N) and/or phosphoethanolamine (pEtN) substituents. Here we use NMR spectroscopy to investigate the attachment sites of the l-Ara4N and pEtN moieties on underivatized, intact EV3 and EV6 and on precursors II(A) and III(A) from kdsA mutants of Salmonella. CDCl(3)/CD(3)OD/D(2)O (2:3:1, v/v) is shown to be a superior solvent for homo- and heteronuclear one- and two-dimensional NMR experiments. The latter were not feasible previously because available solvents caused sample decomposition. Selective inverse decoupling difference spectroscopy is used to determine the attachment sites of substituents on EV3, EV6, II(A), and III(A). l-Ara4N is attached via a phosphodiester linkage to the 4'-phosphates of EV3 and EV6 and has the beta anomeric configuration. pEtN is attached by a pyrophosphate linkage to the 1-phosphate of EV6. The l-Ara4N and pEtN substituents of lipids II(A) and III(A) are attached in the opposite manner, with l-Ara4N on the 1-phosphate of II(A) and pEtN on the 4'-phosphate of III(A). Determination of the proper attachment sites of these substituents is necessary for elucidating the enzymology of lipid A biosynthesis and for characterizing polymyxin-resistant mutants, in which l-Ara4N and pEtN substituents are greatly increased. PMID- 10788471 TI - Molecular and structural properties of three autoimmune IgG monoclonal antibodies to histone H2B. AB - In systemic autoimmune diseases such as lupus the immune system produces autoantibodies to nuclear antigens including DNA and histone molecules. In the present study, we describe three monoclonal IgG antibodies that have been obtained from lupus-prone MRL/lpr mice. These three antibodies react with the amino terminus of histone H2B, a region of the molecule that is accessible in chromatin. Using a series of overlapping H2B synthetic peptides and structural analogues, we have mapped the different epitopes recognized by these antibodies. We have also sequenced the combining sites (variable regions) of the antibodies and modeled their interactions with the corresponding epitopes. Overall, the data suggest that the mechanisms of interaction with antigen are different for each of the three antibodies, even though they all react with the amino-terminal domain of the histone H2B molecule. The results also suggest that the binding between these antibodies and histone H2B is different from that between most antibodies and conventional protein antigens since the heavy chain complementarity determining region 3 appears to play only a limited role in the three antibodies tested. The study of the interaction between self-antigens and spontaneously occurring autoantibodies may help us elucidate the mechanisms driving the expansion of self-reactive lymphocytes. PMID- 10788472 TI - Heparin-binding growth-associated molecule contains two heparin-binding beta sheet domains that are homologous to the thrombospondin type I repeat. AB - Heparin-binding growth-associated molecule (HB-GAM) is an extracellular matrix associated protein implicated in the development and plasticity of neuronal connections of brain. Binding to cell surface heparan sulfate is indispensable for the biological activity of HB-GAM. In the present paper we have studied the structure of recombinant HB-GAM using heteronuclear NMR. These studies show that HB-GAM contains two beta-sheet domains connected by a flexible linker. Both of these domains contain three antiparallel beta-strands. In addition to this domain structure, HB-GAM contains the N- and C-terminal lysine-rich sequences that lack a detectable structure and appear to form random coils. Studies using CD and NMR spectroscopy suggest that HB-GAM undergoes a conformational change upon binding to heparin, and that the binding occurs primarily to the beta-sheet domains of the protein. Search of sequence data bases shows that the beta-sheet domains of HB-GAM are homologous to the thrombospondin type I repeat (TSR). Sequence comparisions show that the beta-sheet structures found previously in midkine, a protein homologous with HB-GAM, also correspond to the TSR motif. We suggest that the TSR sequence motif found in various extracellular proteins defines a beta sheet structure similar to that found in HB-GAM and midkine. In addition to the apparent structural similarity, a similarity in biological functions is suggested by the occurrence of the TSR sequence motif in a wide variety of proteins that mediate cell-to-extracellular matrix and cell-to-cell interactions, in which the TSR domain mediates specific cell surface binding. PMID- 10788473 TI - Calcineurin promotes protein kinase C and c-Jun NH2-terminal kinase activation in the heart. Cross-talk between cardiac hypertrophic signaling pathways. AB - Multiple intracellular signaling pathways have been shown to regulate the hypertrophic growth of cardiomyocytes. Both necessary and sufficient roles have been described for the mitogen activated protein kinase(1) (MAPK) signaling pathway, specific protein kinase C (PKC) isoforms, and calcineurin. Here we investigate the interdependence between calcineurin, MAPK, and PKC isoforms in regulating cardiomyocyte hypertrophy using three separate approaches. Hearts from hypertrophic calcineurin transgenic mice were characterized for PKC and MAPK activation. Transgenic hearts demonstrated activation of c-Jun NH(2)-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK1/2), but not p38 MAPK factors. Calcineurin transgenic hearts demonstrated increased activation of PKCalpha, beta(1), and theta, but not of epsilon, beta(2), or lambda. In a second approach, cultured cardiomyocytes were infected with a calcineurin adenovirus to induce hypertrophy and the effects of pharmacologic inhibitors or co-infection with a dominant negative adenovirus were examined. Calcineurin-mediated hypertrophy was prevented with PKC inhibitors, Ca(2+) chelation, and attenuated with a dominant negative SEK-1 (MKK4) adenovirus, but inhibitors of ERK or p38 activation had no effect. In a third approach, we examined the activation of MAPK factors and PKC isoforms during the progression of load-induced hypertrophy in aortic banded rats with or without cyclosporine. We determined that inhibition of calcineurin activity with cyclosporine prevented PKCalpha, theta, and JNK activation, but did not affect PKCepsilon, beta, lambda, ERK1/2, or p38 activation. Collectively, these data indicate that calcineurin hypertrophic signaling is interconnected with PKCalpha, theta, and JNK in the heart, while PKCepsilon, beta, lambda, p38, and ERK1/2 are not involved in calcineurin mediated hypertrophy. PMID- 10788474 TI - Characterization of Yeast Yea4p, a uridine diphosphate-N-acetylglucosamine transporter localized in the endoplasmic reticulum and required for chitin synthesis. AB - Chitin is an essential cell wall component, synthesis of which is regulated throughout the cell cycle in the yeast Saccharomyces cerevisiae. We cloned an S. cerevisiae gene, YEA4, whose product is homologous to the Kluyveromyces lactis uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) transporter. An epitope tagged Yea4p localized mainly in the 10,000 x g pellet (P2), suggesting endoplasmic reticulum (ER) localization. Membrane vesicles from the P2 fraction showed an 8-fold higher UDP-GlcNAc transport activity in cells harboring a multicopy YEA4 plasmid than in cells harboring vector alone. The activity distribution is identical with the protein distribution in P2, whether the gene is overexpressed or not, suggesting its native localization in P2. Immunolocalization of epitope-tagged Yea4p further revealed ER localization. The increase in transport activity due to the YEA4 overexpression is specific for UDP GlcNAc, but not for UDP-galactose and GDP-mannose. Deltayea4-disrupted cells showed a reduced rate of UDP-GlcNAc transport, contained less chitin, and were larger and rounder in shape than the wild type cells. Our results indicate that YEA4 encodes an ER-localized UDP-GlcNAc transporter that is required for cell wall chitin synthesis in S. cerevisiae. PMID- 10788475 TI - Arginine-rich anti-vascular endothelial growth factor peptides inhibit tumor growth and metastasis by blocking angiogenesis. AB - Tumor angiogenesis is a critical step for the growth and metastasis of solid tumors. Vascular endothelial growth factor (VEGF) is a specific and potent angiogenic factor and contributes to the development of solid tumors by promoting tumor angiogenesis. Therefore, it is a prime therapeutic target for the development of antagonists for treatment of cancer. We identified from peptide libraries arginine-rich hexapeptides that inhibit the interaction of VEGF(165) with VEGF receptor (IC(50) = 2-4 micrometer). They have no effect on binding of basic fibroblast growth factor to cellular receptor. The hexapeptides inhibit the proliferation of human umbilical vein endothelial cells induced by VEGF(165) without toxicity. The peptides bind to VEGF and inhibit binding of both VEGF(165) and VEGF(121), suggesting that the peptides interact with the main body of VEGF but not the heparin-binding domain that is absent in VEGF(121). The identified peptides block the angiogenesis induced by VEGF(165) in vivo in the chick chorioallantoic membrane and the rabbit cornea. Furthermore, one of the hexapeptides, RRKRRR, blocks the growth and metastasis of VEGF-secreting HM7 human colon carcinoma cells in nude mice. Based on our results, the arginine-rich hexapeptides may be effective for the treatment of various human tumors and other angiogenesis-dependent diseases that are related to the action of VEGF and could also serve as leads for development of more effective drugs. PMID- 10788476 TI - Mammalian homologues of yeast sec31p. An ubiquitously expressed form is localized to endoplasmic reticulum (ER) exit sites and is essential for ER-Golgi transport. AB - The yeast coat protein II (COPII) is responsible for vesicle budding from the endoplasmic reticulum (ER). Mammalian functional homologues for all yeast COPII components, except for Sec31p, have been reported. We have cloned a mammalian cDNA whose product (Sec31A) is about 26% identical to Saccharomyces cerevisiae Sec31p. Data base searches also revealed another partial sequence encoding a polypeptide (Sec31B) that is 40% identical to Sec31A. Northern analysis revealed that Sec31A transcripts are ubiquitously and abundantly expressed, while Sec31B transcripts are particularly enriched in the testis and thymus, but present in very low levels in other tissues. Sec31A is localized to vesicular structures that scatter throughout the cell but are concentrated at the perinuclear region. The structures marked by Sec31A contain Sec13, a component of COPII that is well characterized to mark the ER exit sites. Immunoelectron microscopy revealed that Sec31A colocalizes with Sec13 in structures with extensive vesicular-tubular profiles. Antibodies raised against a C-terminal portion of Sec31A co-precipitate Sec13 and inhibit ER-Golgi transport of temperature-arrested vesicular stomatitis G protein in a semi-intact cell assay. Cytosol immunodepleted of Sec31A failed to support vesicular stomatitis G protein transport, which can be rescued by a high molecular weight fraction of the cytosol containing both Sec31A and Sec13. We conclude that Sec31A represents a functional mammalian homologue of yeast Sec31p. PMID- 10788477 TI - Synthesis, 1H NMR structure, and activity of a three-disulfide-bridged maurotoxin analog designed to restore the consensus motif of scorpion toxins. AB - Maurotoxin (MTX) is a 34-residue toxin that has been isolated from the venom of the chactidae scorpion Scorpio maurus palmatus. The toxin displays an exceptionally wide range of pharmacological activity since it binds onto small conductance Ca(2+)-activated K(+) channels and also blocks Kv channels (Shaker, Kv1.2 and Kv1.3). MTX possesses 53-68% sequence identity with HsTx1 and Pi1, two other K(+) channel short chain scorpion toxins cross-linked by four disulfide bridges. These three toxins differ from other K(+)/Cl(-)/Na(+) channel scorpion toxins cross-linked by either three or four disulfide bridges by the presence of an extra half-cystine residue in the middle of a consensus sequence generally associated with the formation of an alpha/beta scaffold (an alpha-helix connected to an antiparallel beta-sheet by two disulfide bridges). Because MTX exhibits an uncommon disulfide bridge organization among known scorpion toxins (C1-C5, C2-C6, C3-C4, and C7-C8 instead of C1-C4, C2-C5, and C3-C6 for three-disulfide-bridged toxins or C1-C5, C2-C6, C3-C7, and C4-C8 for four-disulfide-bridged toxins), we designed and chemically synthesized an MTX analog with three instead of four disulfide bridges ([Abu(19),Abu(34)]MTX) and in which the entire consensus motif of scorpion toxins was restored by the substitution of the two half-cystines in positions 19 and 34 (corresponding to C4 and C8) by two isosteric alpha aminobutyrate (Abu) derivatives. The three-dimensional structure of [Abu(19), Abu(34)]MTX in solution was solved by (1)H NMR. This analog adopts the alpha/beta scaffold with now conventional half-cystine pairings connecting C1-C5, C2-C6, and C3-C7 (with C4 and C8 replaced by Abu derivatives). This novel arrangement in half-cystine pairings that concerns the last disulfide bridge results mainly in a reorientation of the alpha-helix regarding the beta-sheet structure. In vivo, [Abu(19),Abu(34)]MTX remains lethal in mice as assessed by intracerebroventricular injection of the peptide (LD(50) value of 0. 25 microg/mouse). The structural variations are also accompanied by changes in the pharmacological selectivity of the peptide, suggesting that the organization pattern of disulfide bridges should affect the three-dimensional presentation of certain key residues critical to the blockage of K(+) channel subtypes. PMID- 10788478 TI - Endothelial heme oxygenase-1 induction by hypoxia. Modulation by inducible nitric oxide synthase and S-nitrosothiols. AB - The stress protein heme oxygenase-1 (HO-1) is induced in endothelial cells exposed to nitric oxide (NO)-releasing agents, and this process is finely modulated by thiols (Foresti, R., Clark, J. E., Green, C. J., and Motterlini R. (1997) J. Biol. Chem. 272, 18411-18417). Here, we report that up-regulation of HO 1 in aortic endothelial cells by severe hypoxic conditions (pO(2) 24 A on one side of the L2 structure, supporting extensive contact between E1 and L2 domain. Alignment of over 40 lipoyl domain sequences supports Ser(141), Thr(143), and Glu(179) serving as specificity residues for use by E1 from eukaryotic sources. Extensive interactions of the lipoyl-lysine prosthetic group within the active site are supported by the limited inhibition of E1 acetylation of native L2 by L2 domains altered either by mutation of Lys(173) or enzymatic addition of lipoate analogs to Lys(173). Thus, efficient use by mammalian E1 of cognate lipoyl domains derives from unique surface residues with critical interactions contributed by the universal lipoyl-lysine prosthetic group, key specificity residues, and some conserved residues, particularly Asp(172) adjacent to Lys(173). PMID- 10788483 TI - Solution structure of the chitin-binding domain of Bacillus circulans WL-12 chitinase A1. AB - The three-dimensional structure of the chitin-binding domain (ChBD) of chitinase A1 (ChiA1) from a Gram-positive bacterium, Bacillus circulans WL-12, was determined by means of multidimensional heteronuclear NMR methods. ChiA1 is a glycosidase that hydrolyzes chitin and is composed of an N-terminal catalytic domain, two fibronectin type III-like domains, and C-terminal ChBD(ChiA1) (45 residues, Ala(655)-Gln(699)), which binds specifically to insoluble chitin. ChBD(ChiA1) has a compact and globular structure with the topology of a twisted beta-sandwich. This domain contains two antiparallel beta-sheets, one composed of three strands and the other of two strands. The core region formed by the hydrophobic and aromatic residues makes the overall structure rigid and compact. The overall topology of ChBD(ChiA1) is similar to that of the cellulose-binding domain (CBD) of Erwinia chrysanthemi endoglucanase Z (CBD(EGZ)). However, ChBD(ChiA1) lacks the three aromatic residues aligned linearly and exposed to the solvent, which probably interact with cellulose in CBDs. Therefore, the binding mechanism of a group of ChBDs including ChBD(ChiA1) may be different from that proposed for CBDs. PMID- 10788484 TI - Inhibition by extracellular cAMP of phorbol 12-myristate 13-acetate-induced prostaglandin H synthase-2 expression in human pulmonary microvascular endothelial cells. Involvement of an ecto-protein kinase A activity. AB - Exposure of human pulmonary microvascular endothelial cells (HPMECs) to phorbol 12-myristate 13-acetate (PMA) leads to the increase of prostaglandin H synthase (PGHS)-2 protein levels. Under same conditions and according to its constitutive nature, no significant variation of PGHS-1 protein was noted. The elevation of the intracellular cAMP rate is known to enhance PGHS-2 levels through a protein kinase A pathway in various cells. To determine whether the extracellular cAMP also regulates the inducible expression of PGHS, cultured HPMECs were exposed to cAMP alone or in combination with PMA. The PMA-induced PGHS-2 protein was attenuated by the extracellular cAMP. In addition, PGHS-2 activity evaluated through 6-keto-PGF1alpha generation, which was enhanced by PMA was inhibited by extracellular cAMP. Furthermore, in HPMEC medium, PMA-induced PGHS-2 expression was accompanied by the generation of a transferable activity (TA) able to abolish platelet aggregation. This resulting TA was dependent from PGHS-2 pathway, because NS-398, a selective inhibitor of PGHS-2, suppressed its production. The inhibitory TA released by treated HPMECs was also prevented by extracellular cAMP. The specific protein kinase A (PKA) inhibitor blocked the extracellular cAMP effect on both PMA-induced 6-keto-PGF1alpha synthesis and inhibitory TA generation, suggesting the involvement of PKA signaling at the outer surface of HPMECs. Accordingly, we established, in phosphorylation experiments, the presence of an endothelial ecto-protein kinase activity, able to phosphorylate the synthetic substrate kemptide in a cAMP-dependent mode. Reverse transcription polymerase chain reaction analysis showed that PMA-induced PGHS-2 mRNA was markedly reduced by extracellular cAMP. Together, these findings provide the first experimental evidence that extracellular cAMP is able to reduce HPMEC PGHS 2 expression in terms of mRNA, protein, and enzyme activity through an ecto-PKA pathway. In addition, they outline the potential role of endothelial PGHS-2 in the limitation of platelet activation during inflammatory processes. PMID- 10788485 TI - The rabbit antibody repertoire as a novel source for the generation of therapeutic human antibodies. AB - The rabbit antibody repertoire, which in the form of polyclonal antibodies has been used in diagnostic applications for decades, would be an attractive source for the generation of therapeutic human antibodies. The humanization of rabbit antibodies, however, has not been reported. Here we use phage display technology to select and humanize antibodies from rabbits that were immunized with human A33 antigen which is a target antigen for the immunotherapy of colon cancer. We first selected rabbit antibodies that bind to a cell surface epitope of human A33 antigen with an affinity in the 1 nm range. For rabbit antibody humanization, we then used a selection strategy that combines grafting of the complementarity determining regions with framework fine tuning. The resulting humanized antibodies were found to retain both high specificity and affinity for human A33 antigen. PMID- 10788487 TI - Coupled proteolytic and mass spectrometry studies indicate a novel topology for the glycine receptor. AB - Members of the heteropentameric ligand-gated ion channel superfamily rapidly mediate signaling across the synaptic cleft. Sequence analysis and limited experimental studies have yielded a topological model containing four transmembrane alpha-helices, labeled M1 to M4, and a large soluble, extracellular N-terminal domain. This model persists to date despite some recent structural studies that suggest it may be inappropriate. In this study, the topology of the glycine receptor was probed by limited proteolysis coupled to mass spectrometry. Of particular note, accessible cleavage sites within the putative M1 and M3 transmembrane helices were identified. Membrane-associated fragments within the postulated globular extracellular N-terminal domain were also observed. This report presents several key details incorporated in a new topological model and is the first direct experimental evidence that a subset of the transmembrane regions are too short to be membrane-spanning alpha-helices; rather, these regions are proposed to be a mix of alpha-helices and beta-sheets. This report is also the first to exploit the capability of mass spectrometry to probe critically the topology of a class of membrane proteins of unknown structure. PMID- 10788486 TI - Ras/MEK/ERK Up-regulation of the fibroblast KCa channel FIK is a common mechanism for basic fibroblast growth factor and transforming growth factor-beta suppression of myogenesis. AB - The 10T1/2-MRF4 fibroblast/myogenic cell system was used to address the following interrelated questions: whether distinct signaling pathways underlie myogenic inhibition by basic fibroblast growth factor (bFGF) and transforming growth factor (TGF)-beta; which of these pathways also up-regulates the fibroblast intermediate conductance calcium-activated potassium channel, FIK, a positive regulator of cell proliferation; and whether FIK up-regulation underlies some or all myogenic inhibitory signaling events. The results show that myogenic inhibition in 10T1/2-MRF4 cells, by both bFGF and TGF-beta, requires activation of the Ras/mitogen-activated protein (MAP) kinase/MAP kinase-ERK kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway, and resultant FIK up regulation. We show that FIK is instrumental in MEK-dependent suppression of acetylcholine receptor channel expression but that MEK activation and FIK up regulation are not essential to suppression of myosin heavy chain and myotube formation. These data indicate that Ras/MEK/ERK induction of FIK is pivotal to regulation of certain myogenic events by both receptor tyrosine kinases and TGF beta receptor, and this is also the first demonstration of chronic FIK up regulation by the TGF-beta receptor family. Furthermore, the results define the physiologic signaling requirements for growth factor-stimulated FIK up regulation, whereas previous work has concentrated on constitutive FIK up regulation in cells stably transfected with oncoprotein signaling molecules. This study, together with earlier work showing that FIK positively regulates cell proliferation, establishes this member of the IK channel family as a multifunctional, growth factor-regulated signaling molecule. PMID- 10788488 TI - Suppression by metallothionein of doxorubicin-induced cardiomyocyte apoptosis through inhibition of p38 mitogen-activated protein kinases. AB - Cardiomyopathy induced by doxorubicin (DOX) has long been a major impediment of clinical applications of this effective anticancer agent. Previous studies have shown that cardiac-specific metallothionein (MT)-overexpressing transgenic mice are highly resistant to DOX-induced cardiotoxicity. To investigate cellular and molecular mechanisms by which MT participates in this cytoprotection, transgenic mice containing high levels of cardiac MT and non-transgenic controls were treated intraperitoneally with DOX at a single dose of 15 mg/kg and sacrificed on the 4th day after treatment. Myocardial apoptosis was detected by a terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay and confirmed by electron microscopy of immunogold staining of apoptotic nuclei. Dual staining of cardiac alpha-sarcomeric actin using an immunohistochemical method further identified apoptotic myocytes. Apoptosis was significantly inhibited in the transgenic myocardium. The anti-apoptotic effect of MT was further revealed in primary cultures of neonatal mouse cardiomyocytes. Furthermore, DOX activated p38 mitogen-activated protein kinase (MAPK), which was critically involved in the apoptotic process, as demonstrated by inhibition of DOX-induced apoptosis by a p38-specific inhibitor, SB203580. Both DOX-induced p38 MAPK activation and apoptosis were dramatically inhibited in the transgenic cardiomyocytes. The results thus demonstrate that DOX induces apoptosis in cardiomyocytes both in vivo and in vitro and MT suppresses this effect through at least in part inhibition of p38 MAPK activation. PMID- 10788489 TI - Initiation of human DNA replication in vitro using nuclei from cells arrested at an initiation-competent state. AB - Initiation of human DNA replication is investigated in vitro, using initiation competent nuclei isolated from cells arrested in late G(1) phase by a 24-h treatment with 0.5 mm mimosine (Krude, T. (1999) Exp. Cell Res. 247, 148-159). Nuclei isolated from mimosine-arrested HeLa cells initiate semiconservative DNA replication upon incubation in cytosolic extracts from proliferating human cells. Initiation occurs in the absence and presence of a nuclear membrane. The cyclin dependent kinase (Cdk) inhibitors roscovitine and olomoucine inhibit initiation of DNA replication, indicating a dependence of initiation on Cdk activity. Cell fractionation shows that cyclins A, E, and Cdk2 are bound to nuclei from mimosine arrested cells. Exogenously added human cyclin A.Cdk2 and cyclin E.Cdk2 complexes, but not cyclin B1/Cdk1 or cyclin D2/Cdk6, can overcome inhibition of initiation by roscovitine in vitro. Depleting Cdk2 from cytosolic extract does not prevent initiation, demonstrating that cyclin.Cdk2 complexes are not required in the soluble extract, but are provided by the nuclei. Initiation depends further on an essential and soluble activity present in cytosolic extracts from proliferating cells, but not from mimosine-arrested cells, acting together with nuclear cyclin/Cdk2 activity. PMID- 10788490 TI - Directed evolution of beta -glucosidase A from Paenibacillus polymyxa to thermal resistance. AB - The beta-glucosidase encoded by the bglA gene from Paenibacillus polymyxa has a half-life time of 15 min at 35 degrees C and no detectable activity at 55 degrees C. We have isolated random mutations that enhance the thermoresistance of the enzyme. Following a directed evolution strategy, we have combined some of the isolated mutations to obtain a beta-glucosidase with a half-life of 12 min at 65 degrees C, in the range of resistance of thermophilic enzymes. No significant alteration of the kinetic parameters of the enzyme was observed. One of the mutants isolated in the screening for thermoresistant beta-glucosidase had the same resistance to denaturation as the wild type. This mutation caused the accumulation of enzyme in E. coli, probably due to its lower turnover. The structural changes responsible for the properties of the mutant enzymes have been analyzed. The putative causes increasing thermoresistance are as follows: the formation of an extra salt bridge, the replacement of an Asn residue exposed to the solvent, stabilization of the hydrophobic core, and stabilization of the quaternary structure of the protein. PMID- 10788491 TI - Syntaxin 18, a SNAP receptor that functions in the endoplasmic reticulum, intermediate compartment, and cis-Golgi vesicle trafficking. AB - Members of the syntaxin family are target-soluble N-ethylmaleimide-sensitive factor-attachment protein receptors involved in vesicle docking and/or fusion within the exocytic and endocytotic pathways. By using the yeast two-hybrid system, we have identified a novel member of the syntaxin family, syntaxin 18, that binds to alpha-soluble N-ethylmaleimide-sensitive factor-attachment protein. Subcellular fractionation and immunocytochemical analysis revealed that syntaxin 18 is principally located in the endoplasmic reticulum. We examined the effect of overexpression of FLAG-tagged syntaxin 18 and a mutant lacking the N-terminal 81 amino acid residues on protein transport and organelles in the early secretory pathway. Both expressed proteins localized to the endoplasmic reticulum, and the expressed FLAG-syntaxin 18 caused remarkable aggregation of endoplasmic reticulum membranes. Although expression of the FLAG-syntaxin 18 lacking the N-terminal region produced less effect on the morphology of the endoplasmic reticulum, dispersion of the endoplasmic reticulum-Golgi intermediate compartment and cis Golgi was elicited. Moreover, overexpression of the FLAG-syntaxin 18 mutant inhibited protein export from the endoplasmic reticulum. These results taken together suggest that syntaxin 18 functions in transport between the endoplasmic reticulum and Golgi. PMID- 10788492 TI - Phosphorylation of GATA-4 is involved in alpha 1-adrenergic agonist-responsive transcription of the endothelin-1 gene in cardiac myocytes. AB - The expression of endothelin-1 (ET-1) in cardiac myocytes is markedly induced during the development of heart failure in vivo and by stimulation with the alpha(1)-adrenergic agonist phenylephrine in culture. Although recent studies have suggested a role for cardiac-specific zinc finger GATA factors in the transcriptional pathways that modulate cardiac hypertrophy, it is unknown whether these factors are also involved in cardiac ET-1 transcription and if so, how these factors are modulated during this process. Using transient transfection assays in primary cardiac myocytes from neonatal rats, we show here that the GATA element in the rat ET-1 promoter was required for phenylephrine-stimulated ET-1 transcription. Cardiac GATA-4 bound the ET-1 GATA element and activated the ET-1 promoter in a sequence-specific manner. Stimulation by phenylephrine caused serine phosphorylation of GATA-4 and increased its ability to bind the ET-1 GATA element. Inhibition of the extracellularly responsive kinase cascade with PD098059 blocked the phenylephrine-induced increase in the DNA binding ability and the phosphorylation of GATA-4. These findings demonstrate that serine phosphorylation of GATA-4 is involved in alpha(1)-adrenergic agonist-responsive transcription of the ET-1 gene in cardiac myocytes and that extracellularly responsive kinase 1/2 activation plays a role upstream of GATA-4. PMID- 10788493 TI - Export from the endoplasmic reticulum represents the limiting step in the maturation and cell surface expression of the human delta opioid receptor. AB - Synthesis and maturation of G protein-coupled receptors are complex events that require an intricate combination of processes that include protein folding, post translational modifications, and transport through distinct cellular compartments. Relatively little is known about the nature and kinetics of specific steps involved in these processes. Here, the human delta opioid receptor expressed in human embryonic kidney 293S cells is used as a model to delineate these steps and to establish the kinetics of receptor synthesis, glycosylation, and transport. We found that the receptor is synthesized as a core-glycosylated M(r) 45,000 precursor that is converted to the fully mature M(r) 55,000 receptor with a half-time of about 120 min. In addition to trimming and processing of two N-linked oligosaccharides, maturation involves addition of O-glycans containing N acetylgalactosamine, galactose, and sialic acid. In contrast to N-glycosylation, which is initiated co-translationally and is completed when the protein reaches the trans-Golgi network, O-glycosylation was found to occur only after the receptor exits from the endoplasmic reticulum (ER) and was terminated as early as the trans-Golgi cisternae. Once the carbohydrates are fully processed and the receptor reaches the trans-Golgi network, it is transported to the cell surface in about 10 min. The exit from the ER was found to be the limiting step in overall processing of the receptor. This indicates that early events in the folding of the receptor are probably rate-limiting and that receptor folding intermediates are retained in the ER until they can adopt the correct conformation. The overall low efficiency of receptor maturation, less than 50% of the precursor being processed to the fully glycosylated protein, further suggests that only a fraction of the synthesized receptors attain properly folded conformation that allows exit from the ER. This indicates that folding and ER export are key events in control of receptor cell surface expression. Whether or not the low efficiency of the ER export is a general feature among G protein coupled receptors remains to be investigated. PMID- 10788494 TI - Interaction of a kinesin-like calmodulin-binding protein with a protein kinase. AB - Kinesin-like calmodulin-binding protein (KCBP) is a novel member of the kinesin superfamily that is involved in cell division and trichome morphogenesis. KCBP is unique among all known kinesins in having a myosin tail homology-4 region in the N-terminal tail and a calmodulin-binding region following the motor domain. Calcium, through calmodulin, has been shown to negatively regulate the interaction of KCBP with microtubules. Here we have used the yeast two-hybrid system to identify the proteins that interact with the tail region of KCBP. A protein kinase (KCBP-interacting protein kinase (KIPK)) was found to interact specifically with the tail region of KCBP. KIPK is related to a group of protein kinases specific to plants that has an additional sequence between subdomains VII and VIII of the conserved C-terminal catalytic domain and an extensive N-terminal region. The catalytic domain alone of KIPK interacted weakly with the N-terminal KCBP protein but strongly with full-length KCBP, whereas the noncatalytic region did not interact with either protein. The interaction of KCBP with KIPK was confirmed using coprecipitation assays. Using bacterially expressed full-length and truncated proteins, we have shown that the catalytic domain is capable of phosphorylating itself. The association of KIPK with KCBP suggests regulation of KCBP or KCBP-associated proteins by phosphorylation and/or that KCBP is involved in targeting KIPK to its proper cellular location. PMID- 10788495 TI - Gbeta 5gamma 2 is a highly selective activator of phospholipid-dependent enzymes. AB - In this study, Gbeta specificity in the regulation of Gbetagamma-sensitive phosphoinositide 3-kinases (PI3Ks) and phospholipase Cbeta (PLCbeta) isozymes was examined. Recombinant mammalian Gbeta(1-3)gamma(2) complexes purified from Sf9 membranes stimulated PI3Kgamma lipid kinase activity with similar potency (10-30 nm) and efficacy, whereas transducin Gbetagamma was less potent. Functionally active Gbeta(5)gamma(2) dimers were purified from Sf9 cell membranes following coexpression of Gbeta(5) and Ggamma(2-His). This preparation as well as Gbeta(1)gamma(2-His) supported pertussis toxin-mediated ADP-ribosylation of Galpha(i1). Gbeta(1)gamma(2-His) stimulated PI3Kgamma lipid and protein kinase activities at nanomolar concentrations, whereas Gbeta(5)gamma(2-His) had no effect. Accordingly, Gbeta(1)gamma(2-His), but not Gbeta(5)gamma(2-His), significantly stimulated the lipid kinase activity of PI3Kbeta in the presence or absence of tyrosine-phosphorylated peptides derived from the p85-binding domain of the platelet derived-growth factor receptor. Conversely, both preparations were able to stimulate PLCbeta(2) and PLCbeta(1). However, Gbeta(1)gamma(2-His), but not Gbeta(5)gamma(2-His), activated PLCbeta(3). Experimental evidence suggests that the mechanism of Gbeta(5)-dependent effector selectivity may differ between PI3K and PLCbeta. In conclusion, these data indicate that Gbeta subunits are able to discriminate among effectors independently of Galpha due to selective protein-protein interaction. PMID- 10788496 TI - GroEL binds artificial proteins with random sequences. AB - Chaperonin GroEL from Escherichia coli binds to the non-native states of many unrelated proteins, and GroEL-recognizable structural features have been argued. As model substrate proteins of GroEL, we used seven artificial proteins (138 approximately 141 residues), each of which has a unique but randomly chosen amino acid sequence and no propensity to fold into a certain structure. Two of them were water-soluble, and the rest were soluble in 3 m urea. The soluble ones interacted with GroEL in a manner similar to that of a natural substrate; they stimulated the ATPase cycle of GroEL and GroEL/GroES and inhibited GroEL-assisted folding of other protein. All seven artificial proteins were able to bind to GroEL. The results suggest that the secondary structure as well as the specific sequence motif of the substrate proteins are not necessary to be recognized by GroEL. PMID- 10788497 TI - Human homologue of the Drosophila discs large tumor suppressor protein forms an oligomer in solution. Identification of the self-association site. AB - The human homologue of the Drosophila discs large tumor suppressor protein (hDlg), a member of the membrane-associated guanylate kinase (MAGUK) superfamily, interacts with K(+) channels, N-methyl-d-aspartate receptors, calcium ATPase, adenomatous polyposis coli, and PTEN tumor suppressor proteins, and several viral oncoproteins through its PDZ domains. MAGUKs play pivotal roles in the clustering and aggregation of receptors, ion channels, and cell adhesion molecules at the synapses. To investigate the physiological basis of hDlg interactions, we examined the self-association state of full-length hDlg as well as defined segments of hDlg expressed as recombinant proteins in bacteria and insect Sf9 cells. Gel permeation chromatography of full-length hDlg revealed that the purified protein migrates as a large particle of size >440 kDa. Similar measurements of defined domains of hDlg indicated that the anomalous mobility of hDlg originated from its amino-terminal domain. Ultrastructural analysis of hDlg by low angle rotary shadow electron microscopy revealed that the full-length hDlg protein as well as its amino-terminal domain exhibits a highly flexible irregular shape. Further evaluation of the self-association state of hDlg using sedimentation equilibrium centrifugation, matrix-assisted laser desorption/ionization mass spectrometry, and chemical cross-linking techniques confirmed that the oligomerization site of hDlg is contained within its amino terminal domain. This unique amino-terminal domain mediates multimerization of hDlg into dimeric and tetrameric species in solution. Sedimentation velocity experiments demonstrated that the oligomerization domain exists as an elongated tetramer in solution. In vitro mutagenesis was used to demonstrate that a single cysteine residue present in the oligomerization domain of hDlg is not required for its self-association. Understanding the oligomerization status of hDlg may help to explicate the mechanism of hDlg association with multimeric K(+) channels and dimeric adenomatous polyposis coli tumor suppressor protein. Our findings, therefore, begin to rationalize the role of hDlg in the clustering of membrane channels and formation of multiprotein complexes necessary for signaling and cell proliferation pathways. PMID- 10788498 TI - Post-transcriptional regulation of urokinase mRNA. Identification of a novel urokinase mRNA-binding protein in human lung epithelial cells in vitro. AB - We sought to determine if urokinase expression is regulated at the post transcriptional level in cultured lung epithelial cells. We also sought to determine if differences in urokinase expression by cultured human lung carcinoma and non-malignant lung epithelial subtypes were attributable to post transcriptional regulatory mechanisms. Urokinase was expressed by phenotypically diverse lung carcinoma cell lines as well as non-malignant small airway epithelial cells and bronchial epithelial cells. Using gel mobility shift and UV cross-linking assays, we identified a 30-kDa urokinase mRNA-binding protein that selectively bound to a 66-nucleotide protein-binding fragment of urokinase mRNA. The urokinase mRNA-binding protein is found in the cytosolic but not nuclear extracts of non-malignant lung epithelial cells; whereas, it is found in the nuclear but not cytosolic extracts of selected malignant carcinoma-derived cells that express relatively large amounts of urokinase. Chimeric beta globin/urokinase cDNA containing the urokinase mRNA-binding protein binding sequence destabilized otherwise stable beta-globin mRNA. Our results demonstrate that urokinase gene expression in lung epithelial and lung carcinoma-derived cells is regulated at the post-transcriptional level. The mechanism involves an interaction between a 66-nucleotide sequence of the urokinase mRNA 3' untranslated region with a newly recognized urokinase mRNA-binding protein to regulate urokinase mRNA stability. PMID- 10788499 TI - Zinc stoichiometry of yeast RNA polymerase II and characterization of mutations in the zinc-binding domain of the largest subunit. AB - Atomic absorption spectroscopy demonstrated that highly purified RNA polymerase II from the yeast Saccharomyces cerevisiae binds seven zinc ions. This number agrees with the number of potential zinc-binding sites among the 12 different subunits of the enzyme and with our observation that the ninth largest subunit alone is able to bind two zinc ions. The zinc-binding motif in the largest subunit of the enzyme was investigated using mutagenic analysis. Altering any one of the six conserved residues in the zinc-binding motif conferred either a lethal or conditional phenotype, and zinc blot analysis indicated that mutant forms of the domain had a 2-fold reduction in zinc affinity. Mutations in the zinc-binding domain reduced RNA polymerase II activity in cell-free extracts, even though protein blot analysis indicated that the mutant subunit was present in excess of wild-type levels. Purification of one mutant RNA polymerase revealed a subunit profile that was wild-type like with the exception of two subunits not required for core enzyme activity (Rpb4p and Rpb7p), which were missing. Core activity of the mutant enzyme was reduced 20-fold. We conclude that mutations in the zinc binding domain can reduce core activity without altering the association of any of the subunits required for this activity. PMID- 10788500 TI - Overexpression of C-terminal Src kinase blocks 14, 15-epoxyeicosatrienoic acid induced tyrosine phosphorylation and mitogenesis. AB - We have previously reported that 14,15-epoxyeicosatrienoic acid (14, 15-EET) is a potent mitogen for the renal epithelial cell line, LLCPKcl4. This mitogenic effect is dependent upon activation of a protein-tyrosine kinase cascade that results in activation of mitogen-activated protein kinase and phosphatidylinositol 3-kinase. Because of suggestive evidence that 14,15-EET also activated Src in these cells, we stably transfected LLCPKcl4 with an expression construct of the C-terminal Src kinase (CSK), which inhibits Src family kinase activity. In vitro Src kinase activity assays confirmed that in empty vector transfected cells (Vector cells), 14, 15-EET increased Src kinase activity, while in clones overexpressing CSK mRNA and immunoreactive protein (CSK cells), 14,15 EET-induced activation of Src was almost completely blocked (94% inhibition). Of interest, epidermal growth factor (EGF) and fetal bovine serum (FBS) also increased Src activity in Vector cells, but not in CSK cells, further confirming the ability of CSK overexpression to prevent Src activation. CSK cells failed to increase [(3)H]thymidine incorporation in response to exogenous 14,15-EET. In contrast, both EGF and FBS significantly increased [(3)H]thymidine incorporation in CSK cells. Immunoprecipitation with anti-phosphotyrosine antibodies and immunoblotting with an antibody against extracellular signal-regulated kinase (ERK) indicated that in CSK cells, 14,15-EET failed to activate ERK1 and ERK2; however, EGF- and FBS-induced activation of ERKs was not different from that seen in Vector cells. In Vector cells, the 14,15-EET-stimulated tyrosine phosphorylation of ERKs was blocked by pretreatment with 1 microm PP2, a selective inhibitor of Src kinases. The present study demonstrates that 14, 15 EET exerts its mitogenic effects predominantly through a Src kinase-mediated pathway, which is the most upstream signaling step determined to date in the 14,15-EET-activated tyrosine kinase cascade in renal epithelial cells. PMID- 10788501 TI - Arachidonic acid and phosphorylation synergistically induce a conformational change of p47phox to activate the phagocyte NADPH oxidase. AB - The superoxide-producing phagocyte NADPH oxidase can be activated by arachidonic acid (AA) or by phosphorylation of p47(phox) under cell-free conditions. The molecular mechanism underlying the activation, however, has remained largely unknown. Here we demonstrate that AA, at high concentrations (50-100 micrometer), induces direct interaction between the oxidase factors p47(phox) and p22(phox) in parallel with superoxide production. The interaction, being required for the oxidase activation, is mediated via the Src homology 3 (SH3) domains of p47(phox) (p47-(SH3)(2)), which are intramolecularly masked in a resting state. We also show that AA disrupts complexation of p47-(SH3)(2) with its intramolecular target fragment (amino acids 286-340) without affecting association of p47-(SH3)(2) with p22(phox), indicating that the disruption plays a crucial role in the induced interaction with p22(phox). Phosphorylation of p47(phox) by protein kinase C partially replaces the effects of AA; treatment of the SH3 target fragment with PKC in vitro results in a completely impaired interaction with p47-(SH3)(2), and the same treatment of the full-length p47(phox) leads to both interaction with p22(phox) and oxidase activation without AA, but to a lesser extent. Furthermore, phosphorylated p47(phox) effectively binds to p22(phox) and activates the oxidase in the presence of AA at low concentrations (1-5 micrometer), where an unphosphorylated protein only slightly supports superoxide production. Thus AA, at high concentrations, fully induces the interaction of p47(phox) with p22(phox) by itself, whereas, at low concentrations, AA synergizes with phosphorylation of p47(phox) to facilitate the interaction, thereby activating the NADPH oxidase. PMID- 10788502 TI - Norepinephrine induces vascular endothelial growth factor gene expression in brown adipocytes through a beta -adrenoreceptor/cAMP/protein kinase A pathway involving Src but independently of Erk1/2. AB - To identify the signaling pathway that mediates the adrenergic stimulation of the expression of the gene for vascular endothelial growth factor (VEGF) during physiologically induced angiogenesis, we examined mouse brown adipocytes in primary culture. The endogenous adrenergic neurotransmitter norepinephrine (NE) induced VEGF expression 3-fold, in a dose- and time-dependent manner (EC(50) approximately 90 nm). Also, the hypoxia-mimicking agent cobalt, as well as serum and phorbol ester, induced VEGF expression, but the effect of NE was additive to each of these factors, implying that a separate signaling mechanism for the NE mediated induction was activated. The NE effect was abolished by propranolol and mimicked by isoprenaline or BRL-37344 and was thus mediated via beta adrenoreceptors. The NE-induced VEGF expression was fully cAMP mediated, an effect which was inhibited by H-89 and thus was dependent on protein kinase A activity. Involvement of other adrenergic signaling pathways (alpha(1) adrenoreceptors, Ca(2+), protein kinase C, alpha(2)-adrenoreceptors, and pertussis toxin-sensitive G(i)-proteins) was excluded. The specific inhibitor of Src tyrosine kinases, PP2, markedly reduced the stimulation by NE, which demonstrates that a cAMP-dependent Src-mediated pathway is positively connected to VEGF expression. However, inhibition of Erk1/2 MAP kinases by PD98059 was without effect. NE did not prolong VEGF mRNA half-life and its effect was thus transcriptional, and was independent of protein synthesis. These results demonstrate that adrenergic stimulation, through beta-adrenoreceptor/cAMP/protein kinase A signaling, recruits a pathway that branches off from the NE-activated Src-Erk1/2 cascade to enhance transcription of the VEGF gene. PMID- 10788503 TI - Activated 3',5'-cyclic AMP-dependent protein kinase is sufficient to induce neuroendocrine-like differentiation of the LNCaP prostate tumor cell line. AB - Neuroendocrine (NE) differentiation within prostate tumors is proposed to be a contributing factor in disease progression. However, the cellular origin and molecular mechanism controlling differentiation of prostatic NE cells are unresolved. The prostate tumor cell line, LNCaP, can reversibly acquire many NE characteristics in response to treatment with beta-adrenergic receptor agonists and activators of adenylate cyclase. In this study, we demonstrate that these treatments induce protein kinase A (PKA) activation in LNCaP cells and that ectopic expression of a constitutively activated form of the PKA catalytic subunit, CIalpha, results in acquisition of NE characteristics, including the extension of neuritic processes, cessation of mitotic activity, and production of neuron-specific enolase. Forskolin-, epinephrine-, and isoproterenol-dependent NE differentiation of LNCaP cells was significantly inhibited by expressing a dominant negative mutant of the PKA regulatory subunit, RIalpha. These results demonstrate that prostatic NE differentiation in response to these agents depends on PKA activation, and this signaling pathway may provide a therapeutic target for treating advanced forms of prostate cancer. PMID- 10788504 TI - Formation of insoluble oligomers correlates with ST6Gal I stable localization in the golgi. AB - The ST6Gal I is a sialyltransferase that functions in the late Golgi to modify the N-linked oligosaccharides of glycoproteins. The ST6Gal I is expressed as two isoforms with a single amino acid difference in their catalytic domains. The STcys isoform is stably retained in the cell and is predominantly found in the Golgi, whereas the STtyr isoform is only transiently localized in the Golgi and is cleaved and secreted from a post-Golgi compartment. These two ST6Gal I isoforms were used to explore the role of the bilayer thickness mechanism and oligomerization in Golgi localization. Analysis of STcys and STtyr proteins with longer transmembrane regions suggested that the bilayer thickness mechanism is not the predominant mechanism used for ST6Gal I Golgi localization. In contrast, the formation and quantity of Triton X-100-insoluble oligomers was correlated with the stable or transient localization of the ST6Gal I isoforms in the Golgi. Nearly 100% of the STcys and only 13% of the STtyr were found as Triton-insoluble oligomers when Golgi membranes of COS-1 cells expressing these proteins were solubilized at pH 6.3, the pH of the late Golgi. In contrast, both proteins were found in the soluble fraction when these membranes were solubilized at pH 8.0. Analysis of other mutants suggested that a conformational change in the catalytic domain rather than increased disulfide bond-based cross-linking is the basis for the increased ability of STcys protein to form oligomers and the stable localization of STcys protein in the Golgi. PMID- 10788505 TI - Targeted recruitment of histone acetyltransferase activity to a locus control region. AB - Locus control regions (LCRs) are capable of activating target genes over substantial distances and establishing autonomously regulated chromatin domains. The basis for this action is poorly defined. Human growth hormone gene (hGH-N) expression is activated by an LCR marked by a series of DNase I-hypersensitive sites (HSI-III and HSV) in pituitary chromatin. These HSs are located between -15 and -32 kilobases (kb) relative to the hGH transcription start site. To establish a mechanistic basis for hGH LCR function, we carried out acetylation mapping of core histones H3 and H4 in chromatin encompassing the hGH cluster. These studies revealed that the entire LCR was selectively enriched for acetylation in chromatin isolated from a human pituitary somatotrope adenoma and in pituitaries of mice transgenic for the hGH locus, but not in hepatic or erythroid cells. Quantification of histone modification in the pituitary revealed a dramatic peak at HSI/II, the major pituitary-specific hGH LCR determinant (-15 kb), with gradually decreasing levels of modification extending from this site in both 5'- and 3'-directions. The 5'-border of the acetylated domain coincided with the 5' most hGH LCR element, HSV (-34 kb); and the 3'-border included the expressed hGH N gene, but did not extend farther 3' into the placenta-specific region of the gene cluster. These data support a model of LCR function involving targeted recruitment and subsequent spreading of histone acetyltransferase activity to encompass and activate a remote target gene. PMID- 10788506 TI - Multiple growth factor induction of a murine early response gene that complements a lethal defect in yeast ribosome biogenesis. AB - Identification of the transcriptionally activated targets of receptor tyrosine kinases is critical to understanding biologic programs directing both normal and neoplastic growth. To elucidate these molecular processes, we identified genes induced by a potent mesenchymal mitogen, platelet-derived growth factor (PDGF). Using differential display reverse transcription-polymerase chain reaction technology, we isolated a novel growth factor-induced cDNA, San5. San5 transcript induction occurred within 60 min in NIH 3T3 fibroblasts and proceeded in the presence of cycloheximide. Maximal induction of the San5 transcript occurred between 8 and 16 h, concurrent with passage of fibroblasts through G(1). San5 message was potently induced by PDGF AA and BB and acidic and basic fibroblast growth factors, all strong activators of fibroblast proliferation, but not by epidermal growth factor and interleukin-4. In a murine hematopoietic progenitor cell line, San5 transcript induction strictly correlated with [(3)H]thymidine uptake. Isolation and sequencing of the murine San5 cDNA revealed amino acid sequence homology to yeast Nop5p, a nucleolar protein required for pre-rRNA processing and ribosome assembly. Strikingly, SAN5 was able to rescue a nop5 null mutant, implicating SAN5 in the process of ribosome biogenesis. Consistent with this result, SAN5 was localized to the nucleolus in both yeast and mouse. Thus, San5 may provide a link between growth factor receptor activation and the cellular translational machinery. PMID- 10788507 TI - Requirement of SHP2 binding to Grb2-associated binder-1 for mitogen-activated protein kinase activation in response to lysophosphatidic acid and epidermal growth factor. AB - Grb2-associated binder-1 (Gab1) is a multisite docking protein containing a pleckstrin homology (PH) domain, multiple potential tyrosine phosphorylation sites, and several proline-rich sequences. Gab1 becomes tyrosine-phosphorylated in cells stimulated with growth factors, cytokines, and ligands for G protein coupled receptors. A major Gab1-binding protein detected in cells treated with extracellular stimuli is the tyrosine phosphatase, SHP2. Although the role of SHP2-Gab1 interaction in cell signaling has not yet been characterized, SHP2 is known to mediate mitogen-activated protein (MAP) kinase activation induced by the epidermal growth factor (EGF). However, the mechanism by which the SHP2 phosphatase exerts a positive signaling role remains obscure. In this study, we prepared Gab1 mutants lacking the SHP2 binding site (Gab1Y627F), the phosphatidylinositol 3-kinase (PI3K) binding sites (Gab1DeltaPI3K), and the PH domain (Gab1DeltaPH). Expression of Gab1Y627F blocked the extracellular signal regulated kinase-2 (ERK2) activation by lysophosphatidic acid (LPA) and EGF. Conversely, expression of the wild-type Gab1 in HEK293 cells augmented the LPA receptor Edg2-mediated ERK2 activation. Whereas the PH domain was required for Gab1 mediation of ERK2 activation by LPA, it was not essential for EGF-induced ERK2 activation. Expression of Gab1DeltaPI3K had no apparent effect on ERK2 activation by LPA and EGF in the cells that we have examined. These results establish a role for Gab1 in the LPA-induced MAP kinase pathway and clearly demonstrate that Gab1-SHP2 interaction is essential for ERK2 activation by LPA and EGF. These findings also suggest that the positive role of SHP2 in the MAP kinase pathway depends on its interaction with Gab1. PMID- 10788508 TI - Combined action of two transcription factors regulates genes encoding spore coat proteins of Bacillus subtilis. AB - During sporulation of Bacillus subtilis, spore coat proteins encoded by cot genes are expressed in the mother cell and deposited on the forespore. Transcription of the cotB, cotC, and cotX genes by final sigma(K) RNA polymerase is activated by a small, DNA-binding protein called GerE. The promoter region of each of these genes has two GerE binding sites. 5' deletions that eliminated the more upstream GerE site decreased expression of lacZ fused to cotB and cotX by approximately 80% and 60%, respectively but had no effect on cotC-lacZ expression. The cotC lacZ fusion was expressed later during sporulation than the other two fusions. Primer extension analysis confirmed that cotB mRNA increases first during sporulation, followed by cotX and cotC mRNAs over a 2-h period. In vitro transcription experiments suggest that the differential pattern of cot gene expression results from the combined action of GerE and another transcription factor, SpoIIID. A low concentration of GerE activated cotB transcription by final sigma(K) RNA polymerase, whereas a higher concentration was needed to activate transcription of cotX or cotC. SpoIIID at low concentration repressed cotC transcription, whereas a higher concentration only partially repressed cotX transcription and had little effect on cotB transcription. DNase I footprinting showed that SpoIIID binds strongly to two sites in the cotC promoter region, binds weakly to one site in the cotX promoter, and does not bind specifically to cotB. We propose that late in sporulation the rising level of GerE and the falling level of SpoIIID, together with the position and affinity of binding sites for these transcription factors in cot gene promoters, dictates the timing and level of spore coat protein synthesis, ensuring optimal assembly of the protein shell on the forespore surface. PMID- 10788509 TI - Transmembrane domain III plays an important role in ion binding and permeation in the glycine transporter GLYT2. AB - The neuronal glycine transporter GLYT2 takes up glycine from the extracellular space by an electrogenic process where this neurotransmitter is co-transported with sodium and chloride ions. We report in this paper that tyrosine at position 289 of GLYT2a is crucial for ion coupling, glycine affinity and sodium selectivity, stressing the essential role played by this residue of transmembrane domain III in the mechanism of transport. Substitution to tryptophan (Y289W), phenylalanine (Y289F), or serine (Y289S), renders transporters unable to catalyze glycine uptake. Measurements of glycine evoked steady-state currents in transfected HEK-293 cells reveal EC(50) values for glycine 17-fold (Y289F) and 45 fold (Y289S) higher than that of the wild type transporter. Sodium dependence is severely altered in tyrosine 289 mutants, both at the level of apparent affinity and cooperativity, with the more dramatic change corresponding to the less conservative substitution (Y289S). Accordingly, sodium selectivity is gradually lost in Y289F and Y289S mutants, and chloride dependence of glycine evoked currents is markedly decreased in Y289F and Y289S mutants. In the absence of three-dimensional information from these transporters, these results provide experimental evidence supporting the hypothesis of transmembrane domain III being part of a common permeation pathway for substrate and co-transported ions. PMID- 10788510 TI - The fibronectin-binding MSCRAMM FnbpA of Staphylococcus aureus is a bifunctional protein that also binds to fibrinogen. AB - Staphylococcus aureus is an important pathogen capable of causing a wide spectrum of diseases in humans and animals. This bacterium expresses a variety of virulence factors that participate in the process of infection. These include MSCRAMMs (microbial surface components recognizing adhesive matrix molecules) that mediate the adherence of the bacteria to host extracellular matrix components, such as collagen, fibronectin (Fn), and fibrinogen (Fg). Two Fn binding MSCRAMMs, FnbpA and FnbpB, have been previously identified. The Fn binding activity has been localized to the approximately 40-amino acid residue D repeats in the C-terminal part of these proteins. However, no biological activity has yet been attributed to the N-terminal A regions of these proteins. These regions exhibit substantial amino acid sequence identity to the A regions of other staphylococcal MSCRAMMs, including ClfA, ClfB, and SdrG (Fbe), all of which bind Fg. This raises the question of whether the Fn-binding MSCRAMMs can also bind specifically to Fg. In this report, we show that a recombinant form of the A region of FnbpA does specifically recognize Fg. We localize the binding site in Fg for recombinant FnbpA to the gamma-chain, in particular to the C-terminal residues of this polypeptide, the site also recognized by ClfA. In addition, we demonstrate that recombinant FnbpA can compete with ClfA for binding to both immobilized and soluble Fg. By the use of surface plasmon resonance spectroscopy and fluorescence polarization, we determine the dissociation equilibrium constant for the interaction of recombinant FnbpA with intact immobilized Fg and with a synthetic C-terminal gamma-chain peptide, respectively. Finally, by overexpressing FnbpA in a mutant strain of S. aureus that lacks the expression of both ClfA and ClfB, we show that native FnbpA can mediate the interaction of S. aureus with soluble Fg. PMID- 10788511 TI - A role for the p38 MAP kinase pathway in the nuclear shuttling of NFATp. AB - Calcium signals lead to the translocation of nuclear factor of activated T cells (NFAT) from the cytoplasm to the nucleus. This process is regulated by the calcium-activated phosphatase calcineurin, which can be cotransported with NFAT to the nucleus to maintain it transcriptionally active for the duration of calcium signaling. When the calcium signal ceases, NFAT is exported to the cytoplasm, and different NFAT kinases have been reported to oppose calcineurin activities and regulate the nuclear export of NFAT. Here we show that p38 MAPK phosphorylates in vitro and interacts in vivo with NFATp. Furthermore, the activation of this pathway in HeLa cells by cotransfection with activated MKK6 and p38 counteracts the calcium-induced nuclear accumulation of NFATp but not that of NFATc. By contrast, activation of JNK or ERK pathways failed to modify the nuclear shuttling of NFATp. Consistently, activation of p38, but not the JNK MAPK pathway, results in the inhibition of NFATp-driven transcription. In addition, the inhibition of the nuclear accumulation of NFATp by p38 appears to be mediated through the activation of NFATp nuclear export and takes place in a Leptomycin B-sensitive fashion, suggesting the involvement of the exportin CRM1 in this process. Thus, the p38 signal transduction pathway appears to play an important role in the regulation of the nuclear shuttling of NFATp and in cellular homeostasis. PMID- 10788512 TI - Interaction of p55 reverse transcriptase from the Saccharomyces cerevisiae retrotransposon Ty3 with conformationally distinct nucleic acid duplexes. AB - The 55-kDa reverse transcriptase (RT) domain of the Ty3 POL3 open reading frame was purified and evaluated on conformationally distinct nucleic acid duplexes. Purified enzyme migrated as a monomer by size exclusion chromatography. Enzymatic footprinting indicate Ty3 RT protects template nucleotides +7 through -21 and primer nucleotides -1 through -24. Contrary to previous data with retroviral enzymes, a 4-base pair region of the template-primer duplex remained nuclease accessible. The C-terminal portion of Ty3 RT encodes a functional RNase H domain, although the hydrolysis profile suggests an increased spatial separation between the catalytic centers. Despite conservation of catalytically important residues in the RNase H domain, Fe(2+) fails to replace Mg(2+) in the RNase H catalytic center for localized generation of hydroxyl radicals, again suggesting this domain may be structurally distinct from its retroviral counterparts. RNase H specificity was investigated using a model system challenging the enzyme to select the polypurine tract primer from within an RNA/DNA hybrid, extend this into (+) DNA, and excise the primer from nascent DNA. Purified RT catalyzed each of these three steps but was almost inactive on a non-polypurine tract RNA primer. Our studies provide the first detailed characterization of the enzymatic activities of a retrotransposon reverse transcriptase. PMID- 10788513 TI - Identification of the [Fe-S] cluster-binding residues of Escherichia coli biotin synthase. AB - The gene encoding Escherichia coli biotin synthase (bioB) has been expressed as a histidine fusion protein, and the protein was purified in a single step using immobilized metal affinity chromatography. The His(6)-tagged protein was fully functional in in vitro and in vivo biotin production assays. Analysis of all the published bioB sequences identified a number of conserved residues. Single point mutations, to either serine or threonine, were carried out on the four conserved (Cys-53, Cys-57, Cys-60, and Cys-188) and one non-conserved (Cys-288) cysteine residues, and the purified mutant proteins were tested both for ability to reconstitute the [2Fe-2S] clusters of the native (oxidized) dimer and enzymatic activity. The C188S mutant was insoluble. The wild-type and four of the mutant proteins were characterized by UV-visible spectroscopy, metal and sulfide analysis, and both in vitro and in vivo biotin production assays. The molecular masses of all proteins were verified using electrospray mass spectrometry. The results indicate that the His(6) tag and the C288T mutation have no effect on the activity of biotin synthase when compared with the wild-type protein. The C53S, C57S, and C60S mutant proteins, both as prepared and reconstituted, were unable to covert dethiobiotin to biotin in vitro and in vivo. We conclude that three of the conserved cysteine residues (Cys-53, Cys-57, and Cys-60), all of which lie in the highly conserved "cysteine box" motif, are crucial for [Fe-S] cluster binding, whereas Cys-188 plays a hitherto unknown structural role in biotin synthase. PMID- 10788514 TI - Identification of two novel TAF subunits of the yeast Saccharomyces cerevisiae TFIID complex. AB - Using a combination of ion exchange and immunoaffinity chromatography we have purified the general transcription initiation factor TFIID to near homogeneity from Saccharomyces cerevisiae. Yeast TFIID is composed of TBP, the TATA box binding protein, and 14 distinct TBP-associated factors (TAFs), which range in size from 17 to 150 kDa. Twelve of the TAF subunits have been previously identified, but two, TAF48p and TAF65p, are novel. TAF48p exhibits significant sequence similarity to the conserved C-terminal region of Drosophila TAF110p, human TAF130p, and human TAF105p and is encoded by a previously identified gene MPT1. TAF65p shows no significant sequence homology to any previously identified TAFp. The genes encoding TAF48p and TAF65p are single copy and essential for normal yeast cell growth. Furthermore, neither TAF48p nor TAF65p are associated with the histone acetylase Spt-Ada-Gcn5 complex or other non-TFIID TBF.TAF complexes. The significance of these results in terms of TFIID structure, function, and organization is discussed. PMID- 10788515 TI - GIT proteins, A novel family of phosphatidylinositol 3,4, 5-trisphosphate stimulated GTPase-activating proteins for ARF6. AB - ADP-ribosylation factor (ARF) proteins are key players in numerous vesicular trafficking events ranging from the formation and fusion of vesicles in the Golgi apparatus to exocytosis and endocytosis. To complete their GTPase cycle, ARFs require a guanine nucleotide-exchange protein to catalyze replacement of GDP by GTP and a GTPase-activating protein (GAP) to accelerate hydrolysis of bound GTP. Recently numerous guanine nucleotide-exchange proteins and GAP proteins have been identified and partially characterized. Every ARF GAP protein identified to date contains a characteristic zinc finger motif. GIT1 and GIT2, two members of a new family of G protein-coupled receptor kinase-interacting proteins, also contain a putative zinc finger motif and display ARF GAP activity. Truncation of the amino terminal region containing the zinc finger motif prevented GAP activity of GIT1. One zinc molecule was found associated per molecule of purified recombinant ARF GAP1, GIT1, and GIT2 proteins, suggesting the zinc finger motifs of ARF GAPs are functional and should play an important role in their GAP activity. Unlike ARF GAP1, GIT1 and GIT2 stimulate hydrolysis of GTP bound to ARF6. Accordingly we found that the phospholipid dependence of the GAP activity of ARF-GAP1 and GIT proteins was quite different, as the GIT proteins are stimulated by phosphatidylinositol 3,4, 5-trisphosphate whereas ARF-GAP1 is stimulated by phosphatidylinositol 4,5-bisphosphate and diacylglycerol. These results suggest that although the mechanism of GTP hydrolysis is probably very similar in these two families of ARF GAPs, GIT proteins might specifically regulate the activity of ARF6 in cells in coordination with phosphatidylinositol 3-kinase signaling pathways. PMID- 10788516 TI - Identification of a homeodomain binding element in the bone sialoprotein gene promoter that is required for its osteoblast-selective expression. AB - Bone sialoprotein is a 70-kDa extracellular matrix component that is intimately associated with biomineralization, yet the cis-acting elements of the Bsp gene that restrict its expression to mineralizing cells remain uncharacterized. To identify such elements, we analyzed a 2472-base pair fragment of the murine promoter that directs osteoblast-selective expression of a luciferase reporter gene and found that the region between -338 and -178 relative to the transcriptional start is crucial for its osteoblast-selective activity. We identified an element within this region that binds a protein complex in the nuclear extracts of osteoblastic cells and is required for its transcriptional activity. Introduction of a mutation that disrupts a homeodomain binding site within this sequence eliminates both its in vitro binding and nearly all of the osteoblastic-selective activity of the 2472-base pair promoter. We further found that the Dlx5 homeoprotein, which is able to regulate the osteoblast-specific osteocalcin promoter, can bind this element and stimulate its enhancer activity when overexpressed in COS7 cells. These data represent the first description of an osteoblast-specific element within the bone sialoprotein promoter and demonstrate its regulation by a member of a family of factors known to be involved in skeletogenesis. PMID- 10788517 TI - Fibroblast growth factor-2 promotes keratan sulfate proteoglycan expression by keratocytes in vitro. AB - Keratocytes of the corneal stroma produce a specialized extracellular matrix responsible for corneal transparency. Corneal keratan sulfate proteoglycans (KSPG) are unique products of keratocytes that are down-regulated in corneal wounds and in vitro. This study used cultures of primary bovine keratocytes to define factors affecting KSPG expression in vitro. KSPG metabolically labeled with [(35)S]sulfate decreased during the initial 2-4 days of culture in quiescent cultures with low serum concentrations (0.1%). Addition of fetal bovine serum, fibroblast growth factor-2 (FGF-2), transforming growth factor beta, or platelet derived growth factor all stimulated cell division, but only FGF-2 stimulated KSPG secretion. Combined with serum, FGF-2 also prevented serum-induced KSPG down regulation. KSPG secretion was lost during serial subculture with or without FGF 2. Expression of KSPG core proteins (lumican, mimecan, and keratocan) was stimulated by FGF-2, and steady state mRNA pools for these proteins, particularly keratocan, were significantly increased by FGF-2 treatment. KSPG expression therefore is supported by exogenous FGF-2 and eliminated by subculture of the cells in presence of serum. FGF-2 stimulates KSPG core protein expression primarily through an increase in mRNA pools. PMID- 10788518 TI - Retrovirally mediated expression of decorin by macrovascular endothelial cells. Effects on cellular migration and fibronectin fibrillogenesis in vitro. AB - Decorin is a member of the widely expressed family of small leucine-rich proteoglycans. In addition to a primary role as a modulator of extracellular matrix protein fibrillogenesis, decorin can inhibit the cellular response to growth factors. Decorin expression is induced in endothelial cells during angiogenesis, but not when migration and proliferation are stimulated. Thus, decorin may support the formation of the fibrillar pericellular matrix that stabilizes the differentiated endothelial phenotype during the later stages of angiogenesis. Therefore, we tested whether constitutive decorin expression alone could modify endothelial cell migration and proliferation or affect pericellular matrix formation. To this end, replication-defective retroviral vectors were used to stably express bovine decorin, which was detected by Northern and Western blotting. The migration of endothelial cells that express decorin is significantly inhibited in both monolayer outgrowth and microchemotaxis chamber assays. The inhibition of cell migration by decorin was not accompanied by decreased proliferation. In addition, endothelial cells that express decorin assemble an extensive fibrillar fibronectin matrix more rapidly than control cells as assessed by immunocytochemical and fibronectin fibrillogenesis assays. These observations suggest that cell migration may be modulated by the influence of decorin on the assembly of the cell-associated extracellular matrix. PMID- 10788519 TI - Binding of ADAM12, a marker of skeletal muscle regeneration, to the muscle specific actin-binding protein, alpha -actinin-2, is required for myoblast fusion. AB - ADAM12 belongs to the transmembrane metalloprotease ADAM ("a disintegrin and metalloprotease") family. ADAM12 has been implicated in muscle cell differentiation and fusion, but its precise function remains unknown. Here, we show that ADAM12 is dramatically up-regulated in regenerated, newly formed fibers in vivo. In C2C12 cells, ADAM12 is expressed at low levels in undifferentiated myoblasts and is transiently up-regulated at the onset of differentiation when myoblasts fuse into multinucleated myotubes, whereas other ADAMs, such as ADAMs 9, 10, 15, 17, and 19, are expressed at all stages of differentiation. Using the yeast two-hybrid screen, we found that the muscle-specific alpha-actinin-2 strongly binds to the cytoplasmic tail of ADAM12. In vitro binding assays with GST fusion proteins confirmed the specific interaction. The major binding site for alpha-actinin-2 was mapped to a short sequence in the membrane-proximal region of ADAM12 cytoplasmic tail; a second binding site was identified in the membrane-distal region. Co-immunoprecipitation experiments confirm the in vivo association of ADAM12 cytoplasmic domain with alpha-actinin-2. Overexpression of the entire cytosolic ADAM12 tail acted in a dominant negative fashion by inhibiting fusion of C2C12 cells, whereas expression of a cytosolic ADAM12 lacking the major alpha-actinin-2 binding site had no effect on cell fusion. Our results suggest that interaction of ADAM12 with alpha-actinin-2 is important for ADAM12 function. PMID- 10788520 TI - Polyubiquitination of the epidermal growth factor receptor occurs at the plasma membrane upon ligand-induced activation. AB - We have previously shown that, although overexpression of mutant dynamin inhibits clathrin-dependent endocytosis and disrupts high affinity binding of epidermal growth factor (EGF) to the EGF receptor (EGFR), it does not inhibit ligand induced translocation of the EGFR into clathrin-coated pits. In the present study, we demonstrate that, upon ligand binding and incubation at 37 degrees C, the EGFR was polyubiquitinated regardless of overexpression of mutant dynamin. In cells not overexpressing mutant dynamin, the EGFR was rapidly internalized and deubiquitinated. In cells being endocytosis-deficient, due to overexpression of mutant dynamin, however, the EGFR was upon prolonged chase first found in deeply invaginated coated pits, and then eventually moved out of the coated pits and back onto the smooth plasma membrane. Polyubiquitination occurred equally efficiently in cells with or without intact clathrin-dependent endocytosis, while the kinetics of ubiquitination and deubiquitination was somewhat different. We further found that the EGF-induced ubiquitination of Eps15 occurred both in the absence and presence of endocytosis with the same kinetics as polyubiquitination of the EGFR, but that the EGF-induced monoubiquitination of Eps15 was somewhat reduced upon overexpression of mutant dynamin. Our data show that EGF-induced polyubiquitination of the EGFR occurs at the plasma membrane. PMID- 10788521 TI - Modulation of human DNA topoisomerase IIalpha function by interaction with 14-3 3epsilon. AB - Human DNA topoisomerase IIalpha (topo II), a ubiquitous nuclear enzyme, is essential for normal and neoplastic cellular proliferation and survival. Several common anticancer drugs exert their cytotoxic effects through interaction with topo II. In experimental systems, altered topo II expression has been associated with the appearance of drug resistance. This mechanism, however, does not adequately account for clinical cases of resistance to topo II-directed drugs. Modulation by protein-protein interactions represents one mechanism of topo II regulation that has not been extensively defined. Our laboratory has identified 14-3-3epsilon as a topo II-interacting protein. In this study, glutathione S transferase co-precipitation, affinity column chromatography, and immunoprecipitations confirm the authenticity of these interactions. Three assays evaluate the impact of 14-3-3epsilon on distinct topo II functional properties. Using both a modified alkaline comet assay and a DNA cleavage assay, we demonstrate that 14-3-3epsilon negatively affects the ability of the chemotherapeutic, etoposide, to trap topo II in cleavable complexes with DNA, thereby preventing DNA strand breaks. By electrophoretic mobility shift assay, this appears to be due to reduced DNA binding activity. The association of topo II with 14-3-3 proteins does not extend to all 14-3-3 isoforms. No protein interaction or disruption of topo II function was observed with 14-3-3final sigma. PMID- 10788522 TI - V-Type H+-ATPase/synthase from a thermophilic eubacterium, Thermus thermophilus. Subunit structure and operon. AB - V-type ATPase (V(o)V(1)) capable of ATP-driven H(+) pumping and of H(+) gradient driven ATP synthesis was isolated from a thermophilic eubacterium, Thermus thermophilus. When the enzyme was analyzed by gel electrophoresis in the presence of sodium dodecyl sulfate, it showed eight polypeptide bands of which four were subunits of V(1). We also isolated the V(o)V(1) operon, containing nine genes in the order of atpG-I-L-E-X-F-A-B-D, which encoded proteins with molecular sizes of 13, 43, 10, 20, 35, 11, 64, 53, and 25 kDa, respectively. The last four genes were identified as those for V(1) subunits; atpA, B, D, and F encoded the A, B, gamma, and delta subunits, respectively. The first five genes, atpG-atpX, were identified as genes for the V(o) subunits. The product of atpL, the proteolipid subunit, lacked a 19-amino acid presequence and, unlike V-type ATPases, contained two membrane-spanning domains rather than four. The hydrophobic 43-kDa product of atpI is the smallest member so far found of the eukaryotic 100-kDa subunit family. Its electrophoretic band overlapped with the band of the A subunit. Therefore, all the gene products were found in our purified V(o)V(1). We isolated the A(3)B(3) subcomplex reconstituted from the isolated subunits and the A(3)B(3)gamma subcomplex from subunit-expressing Escherichia coli. Electron microscopic observation of these subcomplexes revealed that the gamma subunit of V(1) filled the central cavity of A(3)B(3) and might be central subunit, similar to the gamma subunit of F(1)-ATPase. PMID- 10788523 TI - Caspase-3-dependent and -independent degradation of 28 S ribosomal RNA may be involved in the inhibition of protein synthesis during apoptosis initiated by death receptor engagement. AB - Activation of death receptors initiates intrinsic apoptosis programs in various parts of the cell. To explore the possibility that ribosomal RNA (rRNA), essential for translation in ribosomes, is a target of pro-apoptotic proteins, rRNA was analyzed by electrophoresis in two apoptosis systems: human Jurkat cells treated with anti-Fas antibody and human U937 cells treated with tumor necrosis factor-alpha. In both systems, bands in addition to those of unmodified rRNA were detected a few hours after death receptor engagement. In both systems, the primary additional band was identical and comprised the 3'-terminal region of 28 S rRNA. The degradation of 28 S rRNA was simultaneous with protein synthesis inhibition in both systems. The caspase-3 inhibitor Z-DEVD-FMK suppressed rRNA degradation and protein synthesis inhibition in Jurkat cells but not in U937 cells. Together, our data suggest that different pathways are activated in the two systems we studied, and the final steps in these pathways use very similar or identical ribonucleases to cleave 28 S rRNA. These data suggest a physiological link between rRNA degradation and inhibition of protein synthesis. In general, apoptosis execution initiated by death receptor engagement is promoted by protein synthesis inhibition. Triggered by rRNA degradation, malfunction of the protein synthesis machinery may prompt death execution. PMID- 10788524 TI - Amino acid determinants of alpha 7 nicotinic acetylcholine receptor surface expression. AB - Transient transfection has not been a successful method to express the alpha7 nicotinic acetylcholine receptor such that these receptors are detected on the cell surface. This is not the case for all ligand-gated ion channels. Transient transfection with the 5-hydroxytryptamine type 3 subunit cDNA results in detectable surface receptor expression. Cell lines stably expressing the alpha7 nicotinic acetylcholine receptor produce detectable, albeit variable, levels of surface receptor expression. alpha7 nicotinic acetylcholine receptor surface expression is dependent, at least in part, on cell-specific factors. In addition to factors provided by the cells used for receptor expression, we hypothesize that the surface expression level in transfected cells is an intrinsic property of the receptor protein under study. Employing a set of alpha7-5 hydroxytryptamine type 3 chimeric receptor subunit cDNAs, we expressed these constructs in a transient transfection system and quantified surface receptor expression. We have identified amino acids that control receptor distribution between surface and intracellular pools; surface receptor expression can be manipulated without affecting the total number of receptors. These determinants function independently of the cell line used for expression and the transfection method employed. How these surface expression determinants in the alpha7 nicotinic acetylcholine receptor might influence synaptic efficacy is discussed. PMID- 10788526 TI - A pathogenic 15-base pair deletion in mitochondrial DNA-encoded cytochrome c oxidase subunit III results in the absence of functional cytochrome c oxidase. AB - A 15-base pair, in-frame, deletion (9480del15) in the mitochondrial DNA (mtDNA) encoded cytochrome c oxidase subunit III (COX III) gene was identified previously in a patient with recurrent episodes of myoglobinuria and an isolated COX deficiency. Transmitochondrial cell lines harboring 0, 97, and 100% of the 9480del15 deletion were created by fusing human cells lacking mtDNA (rho(0) cells) with platelet and lymphocyte fractions isolated from the patient. The COX III gene mutation resulted in a severe respiratory chain defect in all mutant cell lines. Cells homoplasmic for the mutation had no detectable COX activity or respiratory ATP synthesis, and required uridine and pyruvate supplementation for growth, a phenotype similar to rho(0) cells. The cells with 97% mutated mtDNA exhibited severe reductions in both COX activity (6% of wild-type levels) and rates of ATP synthesis (9% of wild-type). The COX III polypeptide in the mutant cells, although translated at rates similar to wild-type, had reduced stability. There was no evidence for assembly of COX I, COX II, or COX III subunits in a multisubunit complex in cells homoplasmic for the mutation, thus indicating that there was no stable assembly of COX I with COX II in the absence of wild-type COX III. In contrast, the COX I and COX II subunits were assembled in cells with 97% mutated mtDNA. PMID- 10788525 TI - Processing and maturation of flavocytochrome b558 include incorporation of heme as a prerequisite for heterodimer assembly. AB - The phagocyte NADPH-dependent oxidase generates superoxide by reducing molecular oxygen through a transmembrane heterodimer known as flavocytochrome b(558) (flavocytochrome b). We investigated the biosynthesis of flavocytochrome b subunits gp91(phox) and p22(phox) to elucidate features of flavocytochrome b processing in myeloid cells. Although the gp91(phox) precursor, gp65, was processed to gp91(phox) within 4-8 h of chase, unassembled gp65 and p22(phox) monomers were degraded by the cytosolic proteasome. gp65 associated with p22(phox) post-translationally, within 1-4 h of chase, but prior to its modification in the Golgi complex. Moreover, p22(phox) coprecipitated with unglycosylated gp91(phox) primary translation product made in the presence of tunicamycin, suggesting that heterodimer formation does not require glycosylation. Blocking heme synthesis with succinyl acetone completely inhibited heterodimer formation, although biogenesis of gp65 and p22(phox) was unaffected. In succinyl acetone-treated cells, p22(phox) and gp65 were degraded completely by 8 h of chase, a process mediated by the cytosolic proteasome. Taken together, these data suggest that the formation of the gp65-p22(phox) heterodimer is relatively inefficient and that acquisition of heme by gp65 precedes and is required for its association with p22(phox), a process that requires neither the addition of N-linked oligosaccharides nor modification in the Golgi complex. PMID- 10788527 TI - Vasculitis-aims of therapy. An overview. PMID- 10788528 TI - Cost-effectiveness of non-steroidal anti-inflammatory drugs (NSAIDs)-what makes a NSAID good value for money? PMID- 10788529 TI - Sjogren's syndrome-time for a new approach. PMID- 10788530 TI - How to improve morbidity and mortality in systemic lupus erythematosus. PMID- 10788531 TI - Diagnosis and evaluation of vasculitis. PMID- 10788533 TI - Analysis of p53 tumour suppressor gene somatic mutations in rheumatoid arthritis synovium. AB - OBJECTIVE: In order to study the role of the p53 tumour suppressor gene in the proliferation of rheumatoid arthritis (RA) synovium, we analysed the mutation of p53 in the synovial fibroblast-like type B synoviocyte from RA patients. METHODS: Synovial fibroblast-like type B synoviocytes were prepared from the synovial tissues from nine Japanese patients with RA. The p53 cDNA region from exons 4-11 was screened for mutations by the streamlined mutation detection method in which polymerase chain reaction (PCR) products are post-labelled and are analysed by automated capillary electrophoresis using single-strand conformation polymorphism conditions, followed by direct sequencing of the subclones of the PCR products. RESULTS: p53 mutation with possible functional alteration was detected in four of the nine RA patients (44.4%). Of a total of 262 p53 cDNA subclones, 10 subclones were carrying 10 p53 mutations, eight of which were associated with amino acid alterations or protein truncation. Of the p53 functional mutations, a substitution of Gly at amino acid residue 245 to Asp (G245D) was identified in two patients in three subclones. G245D was the first mutation that was recurrently identified in different RA individuals. G245D is also one of the relatively common mutations in human cancers. CONCLUSIONS: In some patients with RA, dysfunction of p53 might play a role in the proliferation of the synovial tissue. G245D mutation might especially need further study as it is the first recurrently identified p53 mutation in RA and is also one of the frequently identified mutations in human cancers. PMID- 10788532 TI - Rapid lupus autoantigen relocalization and reactive oxygen species accumulation following ultraviolet irradiation of human keratinocytes. AB - OBJECTIVE: In vitro treatment with ultraviolet B (UVB) induces relocalization of lupus autoantigens to the cell surface. We have addressed the relationship between autoantigen relocalization, accumulation of intracellular reactive oxygen species (ROS) and the induction of apoptosis following UVA and UVB exposure. METHODS: Human primary keratinocytes were exposed in vitro to doses of UVA and UVB equivalent to 0.01-4 times the minimal erythemal dose. The cellular locations of Ro60, Ro52, Sm, U2-B" and La were determined using monoclonal antibodies. ROS accumulation and apoptosis induction were assessed using the intracellular ROS probe 2'7'-dichlorodihydrofluorescein diacetate, and the viability stains Hoechst 33342 and propidium iodide. RESULTS: UV treatment induced the relocalization of all five autoantigens investigated and an accumulation of ROS. UVA and UVB induced necrosis and apoptosis, respectively. CONCLUSION: These data suggest that both UVA and UVB induce ROS within keratinocytes but have significantly different effects upon autoantigen relocalization and cell viability. PMID- 10788535 TI - What determines arthritogenicity of bacterial cell wall? A study on Eubacterium cell wall-induced arthritis. AB - OBJECTIVE: To study what determines the arthritogenicity of the bacterial cell wall (CW) using Eubacterium CW-induced arthritis in the rat. METHODS: Eubacterium aerofaciens, previously reported as arthritogenic, and E. limosum and E. alactolyticum, known as non-arthritogenic, were used. Gas chromatography-mass spectrometry (GC-MS) was applied to analyse the chemical composition of the bacterial cell wall. Cellular immune response was measured by concanavalin A (Con A) stimulation and FACScan analysis. Also, serum antibodies against the injected cell wall were determined. RESULTS: Unexpectedly, from the two strains of E. aerofaciens used only one proved to be arthritogenic (with a CW inducing chronic arthritis after a single intraperitoneal injection), even though these two strains were 100% identical by 16S rDNA analysis. CW of the other E. aerofaciens strain induced only transient acute arthritis; CW of E. limosum and E. alactolyticum induced weak signs of acute arthritis. Based on the GC-MS analysis and on the results published previously, putative structures of peptidoglycan (PG) in the four CW preparations are presented. It is apparent that the presence of lysine in position 3 of the PG stem peptide contributes to arthritogenicity but is alone not decisive. Both strains of E. aerofaciens were immunosuppressive, when tested by Con A response at 2 weeks after CW injection. Such an immunosuppression was not observed after injection of CW from E. limosum or E. alactolyticum. FACScan analysis for six T cell markers and studies on serum antibody responses did not reveal any differences in the effect of the four bacterial strains used. CONCLUSIONS: The results obtained suggest that the chemical structure of PG present in the bacterial CW is decisive in determining arthritogenicity/non-arthritogenicity. Therefore, from two bacterial strains belonging to normal human intestinal flora and 100% identical by 16S rDNA analysis, one proved to be arthritogenic and the other non-arthritogenic. PMID- 10788534 TI - Familial vs sporadic rheumatoid arthritis (RA). A prospective study in an early RA inception cohort. AB - OBJECTIVES: To study potential differences in demographic, process and outcome variables between familial and sporadic rheumatoid arthritis (RA) in an early RA inception cohort. METHODS: In 1998, we ascertained the familial status of all collaborative patients in a large early RA inception cohort at our department. Familial RA was defined by the presence of at least two siblings fulfilling the American College of Rheumatology criteria for RA. Baseline demographic data and prospectively recorded disease activity variables, therapies and radiological damage during the first 6 yr of disease were included in the analysis. A regression analysis was performed to assess whether familial clustering is a prognostic factor. RESULTS: We identified 142 patients with sporadic and 36 with familial RA. The most striking difference between these groups was the larger sibship size in multicase families (8.2 +/- 2.5 vs 5. 5 +/- 2.8; P < 0.0001). Age at onset was similar in both groups, although males with familiar RA were younger at disease onset than those with sporadic RA (median 50 vs 57 yr; P=0.03). No differences were found in gender, presence of rheumatoid factor (RF), antinuclear factor and HLA-DR typing or in disease activity, interventions and outcome over 6 yr of follow-up. Early radiological damage and disease activity, but not familial history of RA were prognostic for X-ray damage. CONCLUSION: We show that sibship size is the only relevant risk factor for familial RA. No differences in genotypic and phenotypic characteristics, disease severity or radiological damage were observed among familial and sporadic RA. Familial history of RA is not a poor prognostic factor. This prospective study confirms previous cross-sectional findings in the Dutch population. PMID- 10788536 TI - The incidence and clinical characteristics of peripheral arthritis in polymyalgia rheumatica and temporal arteritis: a prospective study of 231 cases. AB - OBJECTIVE: To evaluate the incidence and characteristics of peripheral arthritis in polymyalgia rheumatica and temporal arteritis, and to ascertain the incidence of rheumatoid arthritis among such cases. Patients and methods. In total, 231 patients were selected from a prospective population-based study. All patients were clinically examined on several occasions and followed until cessation of therapy and permanent disease remission. RESULTS: Of the 231 cases, 38.5% presented peripheral arthritis either at diagnosis or during the disease course. At diagnosis, peripheral arthritis was not observed among patients with temporal arteritis. Peripheral arthritis occurring during the disease course was more often polyarticular and needed additional treatment more frequently than joint inflammation presenting at diagnosis. Only one case had distal pitting oedema. Rheumatoid arthritis developed in 4.8% of the cases and exclusively among patients with polymyalgia rheumatica. CONCLUSION: Aetiopathogenic differences may exist between polymyalgia rheumatica and temporal arteritis as peripheral arthritis and the development of rheumatoid arthritis were observed among the former patient group only. PMID- 10788537 TI - Radiographic joint space in rheumatoid glenohumeral joints. A 15-year prospective follow-up study in 74 patients. AB - OBJECTIVE: To evaluate radiographically the glenohumeral (GH) joint space in patients with long-term rheumatoid arthritis (RA). METHODS: A cohort of 74 patients with RA were followed prospectively for 15 yr. At the end point, 148 shoulders were radiographed using a standard method. The GH joint space was examined from the radiographs using a method developed previously for population studies; the joint space was measured at three different sites and the average of the three measurements, the integral space, was calculated. Destruction of the GH joints was assessed with the Larsen method on a scale of 0-5 and compared with the joint space measurements. RESULTS: The mean GH joint space in RA patients was 3. 1 (S.D. 3.3), range -17.3 to 5.7 mm; 2.7 mm (S.D. 4.5) in men and 3. 2 mm (S.D. 2.8) in women. The mean of the affected joints (Larsen grades 2-5), 1.7 mm (S.D. 4.5), was notably narrower than the mean 4. 4 mm (S.D. 0.6) of the non affected (Larsen grades 0-1) joints. Pathological GH joint space, less than 2 mm, was found in five (15%) of 36 joints in men and in 14 (13%) of 112 joints in women. All the joints graded as Larsen 4 and 5 (n = 17) fulfilled this pathological criterion. Joint space narrowing was associated [r = - 0.66, 95% confidence interval (CI): -0.56 to -0.75] with increasing destruction (Larsen grading) of the joint. The narrowing was significant between non- (Larsen 0, 1), moderately (Larsen 2, 3) and severely (Larsen 4, 5) affected joints (P < 0.001). However, a remarkable step in this process occurred between Larsen grades 3 and 4 when the mean joint space diminished from 3.1 to 0.3 mm. CONCLUSIONS: Joint space narrowing is a frequent consequence of GH joint rheumatoid affection. However, joint space narrowing is a late phenomenon occurring not until after marked erosive destruction, which should be noted when using the Larsen method for GH joints. PMID- 10788538 TI - Effects of creatine supplementation on muscle weakness in patients with rheumatoid arthritis. AB - BACKGROUND AND OBJECTIVES: Patients with rheumatoid arthritis (RA) frequently suffer from muscle weakness. Oral administration of creatine has been shown to improve muscle strength in healthy subjects. The objective of this study was to examine the effect of oral creatine supplementation on muscle weakness, disease activity and activities of daily living in patients with RA. METHODS: During a period of 3 weeks, 12 patients with RA were treated with creatine monohydrate (20 g/day for 5 days followed by 2 g/day for 16 days). They were examined on entry and at the end of the study. The patients were investigated clinically, blood and urine samples were obtained, muscle biopsies were performed before and after treatment, muscle strength was determined, and self-administered patient questionnaires were completed. RESULTS: From all patients we were able to obtain full clinical and questionnaire data, while biopsies were taken from 12 patients at the start and from nine patients at the end of the study. Muscle strength, as determined by the muscle strength index, increased in eight of 12 patients. In contrast, physical functional ability and disease activity did not change significantly. The creatine concentration in serum and skeletal muscle increased significantly, while creatine phosphate and total creatine did not increase in skeletal muscle. The skeletal muscle creatine content was associated with muscle strength at baseline but not after administration of creatine. The changes in muscle strength were not associated with the changes in skeletal muscle creatine or creatine phosphate. CONCLUSION: Although the skeletal muscle creatine content and muscle strength increased with creatine administration in some patients with RA, a clear clinical benefit could not be demonstrated for this treatment when the patients were considered as one group. PMID- 10788539 TI - Linkage analysis of candidate genes as susceptibility loci for osteoarthritis suggestive linkage of COL9A1 to female hip osteoarthritis. AB - OBJECTIVE: To examine 11 candidate genes as susceptibility loci for osteoarthritis (OA). METHODS: A total of 481 families have been ascertained in which at least two siblings have had joint replacement surgery of the hip, or knee, or hip and knee for idiopathic OA. Each candidate gene was targeted using one or more intragenic or closely linked microsatellite marker. The linkage data were analysed unstratified and following stratification by sex and by joint replaced (hip or knee). RESULTS: The analyses revealed suggestive linkage of the type IX collagen gene COL9A1 (6q12-q13) to a subset of 132 families that contained affected females who were concordant for hip OA (female-hip) with a P value of 0.00053 and logarithm of the odds (LOD) score of 2.33 [corrected P-value of 0. 0016, corrected LOD score of 1.85]. CONCLUSIONS: COL9A1 may therefore be a susceptibility locus for female hip OA. In addition, there was weak evidence of linkage to HLA/COL11A2 (6p21.3) in female hip OA with a corrected P-value of 0.016. PMID- 10788540 TI - Bone mineral density in adults with Marfan syndrome. AB - OBJECTIVES: Reduced bone mineral density (BMD) has been reported in pre menopausal women and children with Marfan syndrome (MFS). The bone mineral status of adult men with MFS is unknown. The objective of this study was to determine the BMD of adult men and women with MFS. METHODS: BMD (g/cm(2)) was measured by dual-energy X-ray absorptiometry (DXA) of the lumbar spine (L1-L4), femoral neck and total hip in 25 adults (12 male) who fulfilled the 1996 MFS diagnostic criteria. The results were compared with age- and sex-matched controls and expressed as S.D. from the population mean (z score). RESULTS: Overall, BMD was significantly reduced in the lumbar spine (z = -0.42 +/- 0.97, P < 0.05), total hip (z = -0.57 +/- 0.88, P < 0.005) and femoral neck (z = -0.51 +/- 0.88, P < 0. 005). In women alone, BMD was reduced at the femoral neck (z = -0.53 +/- 0.95, P < 0.05) and at the hip (z = -0.64 +/- 0.77, P < 0.005). In men, BMD was reduced at the femoral neck (z = -0.48 +/- 0.84, P < 0.05) with a non-significant trend to lower BMD at the hip (z = -0. 49 +/- 1.01, P = 0.054) and lumbar spine (z = 0.59 +/- 1.02, P = 0. 09). CONCLUSION: Axial BMD is lower than normal in Marfan adults. This reduction may contribute to fractures seen in the Marfan population. PMID- 10788541 TI - A comparison of three methods for measuring thoracic kyphosis: implications for clinical studies. AB - OBJECTIVES: To compare the Cobb technique for measuring kyphosis with an alternative Cobb method and a computer-assisted curve assessment technique, and to examine the influence of vertebral body and disc shape on kyphosis. METHODS: Kyphosis measurements were derived from 93 lateral spinal radiographs or sagittal computed tomography images of cadaveric spines, using: (i) a computer-assisted method for estimating radius of curvature; (ii) the traditional Cobb method; and (iii) an alternative Cobb method. Regression models were applied for agreement analyses, and to examine the relative contribution of vertebral body and disc shape on the extent of curvature. Results and conclusions. Strong associations existed between curvature and angle data derived from the three methods, confirming the clinical utility of these techniques for the quantification of thoracic kyphosis. However, the traditional Cobb method tended to overestimate kyphosis in the presence of vertebral body end-plate deformation. The degree of kyphosis was strongly reflective of the extent of deformity of the vertebral bodies, and to a lesser extent the shape of the thoracic discs. PMID- 10788542 TI - A clinical study of CPH 82 vs methotrexate in early rheumatoid arthritis. AB - OBJECTIVES: The objectives of this study were to evaluate the therapeutic efficacy of CPH 82 in comparison with methotrexate (MTX) in adult patients with early, active rheumatoid arthritis (RA) and to compare the tolerance and safety profiles of the two drugs. METHODS: The study was a 24-week, double-blind, randomized study in 10 centres of 100 patients with active RA, with a disease duration of less than 2 yr at the start of treatment, which consisted of either CPH 82 300 mg/day or MTX 10 mg/week. The six primary effect variables were: number of swollen joints, Ritchie's articular index, patient's pain score, patient's global score, Health Assessment Questionnaire (HAQ) and C-reactive protein (CRP). Erythrocyte sedimentation rate (ESR), physician's global score and the efficacy according to the European League Against Rheumatism (EULAR) and American College of Rheumatology (ACR) response criteria were also analysed. RESULTS: There was a significant improvement for both drugs in all variables. Significant differences between the drugs in favour of MTX were found only in patient's pain score, CRP and ESR. By the EULAR criteria, 76% and 86% were judged to be responders in the CPH 82 and MTX groups, respectively. By the ACR criteria, the corresponding figures were 58% and 64%. The most common side-effects were gastrointestinal, which were similar in both groups. The numbers of treatment failures due to adverse events were two with CPH 82 and 14 with MTX. CONCLUSIONS: The clinical effect of CPH 82 in this study was comparable to that of MTX at a dose of 10 mg/week. Both drugs reduced acute-phase reactants, MTX more effectively than CPH 82. The safety profile of CPH 82 was more favourable than that of MTX without folic acid supplementation. PMID- 10788544 TI - Vasculitis-aims of therapy meeting report, cambridge, 10-11 september 1999 PMID- 10788543 TI - Feasibility and validity of the RADAI, a self-administered rheumatoid arthritis disease activity index. AB - OBJECTIVE: The goal of the Rheumatoid Arthritis Disease Activity Index (RADAI) is to provide an easy to use assessment of disease activity. It is a self administered questionnaire that combines five items into a single index: current and past global disease activity, pain, morning stiffness and a joint count. METHODS: A sample of 484 rheumatoid arthritis (RA) patients was used to assess the internal consistency and the convergent validity of the RADAI. This was achieved by calculating Cronbach's alpha and RADAI item and total score correlations with core set measures and DAS28. RESULTS: Cronbach's alpha was 0.87, supporting the summation of the items into a single index. The index correlated best with physicians' global assessment (r = 0.59; P < 0.0001), the Health Assessment Questionnaire (r = 0.55; P < 0.0001) and the number of tender joints (r = 0.55; P < 0.0001). Correlation with the erythrocyte sedimentation rate was low (r = 0.27; P < 0.0001). The RADAI and the DAS28 were correlated (r = 0.53; P < 0.0001), but there was low agreement. CONCLUSIONS: The RADAI is valid to assess disease activity in RA patients. However, the RADAI may not automatically replace other measures of disease activity, such as the DAS28. PMID- 10788546 TI - Re: Harms et al. Forces measured during spinal manipulative procedures in two age groups. PMID- 10788545 TI - Regression of calcinosis associated with adult dermatomyositis following diltiazem therapy. PMID- 10788547 TI - Coeliac disease can still present with osteomalacia! PMID- 10788548 TI - A fibromyalgia scale in a general rheumatology clinic. PMID- 10788549 TI - Cold agglutinins and cryoglobulins in a patient with acute aortoarteritis (Takayasu's disease) and tuberculous lymphadenitis. PMID- 10788550 TI - Septic arthritis due to Streptococcus bovis as presenting sign of 'silent' colon carcinoma. PMID- 10788551 TI - Complete remission of relapsing eosinophilic fasciitis associated with bronchial asthma following regular steroid inhalation. PMID- 10788552 TI - The ARC steroid trial and its effects on clinical practice. PMID- 10788553 TI - Fritz Haber: 1868-1934. PMID- 10788554 TI - Reevaluation of the cancer potency factor of toxaphene: recommendations from a peer review panel. AB - This reevaluation of the current U.S. EPA cancer potency factor for toxaphene is based upon a review of toxaphene carcinogenesis bioassays in mice conducted by Litton Bionetics (unpublished report, 1978) and the National Cancer Institute (NCI) (Technical Report Series No. 37, conducted by Gulf South Research Institute, 1979). The mechanistic data available for toxaphene, including consideration of the potential of the compound to induce genotoxicity, was examined with an emphasis on whether this information supports a change in the cancer potency factor. If a quantitative dose-response assessment for toxaphene is to be performed, the data from both the NCI and Litton cancer bioassays should be used. Additionally, liver tumor results from female mice, rather than male mice, should be used for estimating potential human cancer risk because the background rate of liver tumors in females is lower and less variable than that exhibited by males. An ED(10) was estimated as the point of departure. The mechanistic data were not sufficient to fully support a margin of exposure approach. Therefore, we believe that applying a linear extrapolation from the ED(10) to the origin is an appropriate means to estimate risk at low doses. This is a highly conservative approach and, when it is applied, we conclude that the current EPA cancer potency factor should be reduced from 1.1 (mg/kg/day)(-1) to 0.1 (mg/kg/day)(-1). PMID- 10788555 TI - Is the human carcinogen arsenic carcinogenic to laboratory animals? PMID- 10788556 TI - Intratracheal instillation as an exposure technique for the evaluation of respiratory tract toxicity: uses and limitations. AB - The evaluation of respiratory tract toxicity from airborne materials frequently involves exposure of animals via inhalation. This provides a natural route of entry into the host and, as such, is the preferred method for the introduction of toxicants into the lungs. However, for various reasons, this technique cannot always be used, and the direct instillation of a test material into the lungs via the trachea has been employed in many studies as an alternative exposure procedure. Intratracheal instillation has become sufficiently widely used that the Inhalation Specialty Section of the Society of Toxicology elected to develop this document to summarize some key issues concerning the use of this exposure procedure. Although there are distinct differences in the distribution, clearance, and retention of materials when administered by instillation compared to inhalation, the former can be a useful and cost-effective procedure for addressing specific questions regarding the respiratory toxicity of chemicals, as long as certain caveats are clearly understood and certain guidelines are carefully followed. PMID- 10788557 TI - Ethane sulfonate metabolite of alachlor: assessment of oncogenic potential based on metabolic and mechanistic considerations. AB - Chronic administration of alachlor has been shown to produce neoplastic responses in the nasal turbinate mucosa, glandular stomach mucosa, and thyroid follicular epithelium of rats. Subsequent studies have shown that specific metabolic activation of alachlor is required for nasal tumor formation, and that non genotoxic, threshold-sensitive processes produce all three tumors. The herbicide alachlor is degraded in the soil by microbial action to the tertiary ethane sulfonate metabolite (ESA). The acute and subchronic toxicity of ESA is very low, and the metabolite did not produce developmental toxicity or genotoxicity. The studies described here were conducted to determine whether ESA shares a common mechanism of oncogenicity with alachlor in rats. Specifically, we studied ESA's pharmacokinetics and ability to produce changes that are causally associated with the oncogenicity of alachlor. These studies demonstrated that ESA was poorly absorbed and underwent minor metabolism, which contrasted with the significant absorption and substantial metabolism observed with alachlor. ESA was also excreted more quickly than alachlor and showed no evidence of accumulation in the nasal turbinates, a site of oncogenicity for alachlor in the rat. In addition, ESA did not elicit the characteristic preneoplastic changes observed in the development of alachlor-induced nasal, stomach, and thyroid tumors. The results of these studies support the conclusion that ESA does not share a common oncogenic mechanism with alachlor and would not be expected to produce the same oncogenic responses observed following chronic alachlor exposure in rats. PMID- 10788558 TI - Dose response of early effects related to tumor promotion of 2 acetylaminofluorene. AB - The genotoxic effects of 2-acetylaminofluorene (AAF) alone cannot explain the formation of rat liver tumors. It has been proposed that mitochondrial effects are associated with its tumor-promoting properties. These mitochondrial effects are thought to trigger apoptosis and regenerative proliferation, which alters the liver lobule in a cirrhosis-like manner. A situation is generated which favors the growth of initiated cells. To test this sequence of events, the dose dependence of early effects with time was studied. Male Wistar rats received 50, 100, 200, 400, or 800 ppm AAF in the diet and the following endpoints were analyzed at 2, 4, 8, and 16 weeks of feeding: apoptotic cell death, cell proliferation, GST-P-positive foci (placental form of glutathione S-transferase), and morphological alterations. Hydrolyzable hemoglobin adducts were used as a dosimeter for metabolic activation after 2 weeks of feeding. The hemoglobin adduct levels increased linearly with dose. With the conventional carcinogenic concentration of 200-ppm AAF in the diet, the number of apoptoses increased first, predominantly in the periportal area (2 weeks). Cell proliferation followed with some delay (4 weeks). This reflects regenerative tissue response and not the growth of initiated cells, because the number of enzyme-altered foci was still extremely low at that time. Foci developed only later when the morphology had changed. With 50 ppm AAF in the diet, a no-effect level had not been reached for any of the endpoints, but foci developed much more gradually than with higher doses. Unexpectedly, the proliferative response stabilized at 8 weeks with a labeling index of 12-17, with all AAF concentrations. The observed sequence of events supports the hypothesis. It is concluded that (1) The proliferation of initiated cells-defined as promotable cells-is largely determined by the cellular environment, such as morphologically altered liver. (2) The morphological alterations in rat liver result from imperfect regeneration from toxic effects. (3) Imperfect regeneration results from limitations in the possibilities to adapt to chemical stress. (4) If these limits are overwhelmed and morphology has changed to a certain extent, preneoplastic foci develop; this occurs much faster, at least, than without these changes. PMID- 10788559 TI - Gabapentin-induced mitogenic activity in rat pancreatic acinar cells. AB - Gabapentin induces pancreatic acinar cell tumors in rats through unknown, yet apparently nongenotoxic mechanisms. The primary objective of this study was to determine whether gabapentin acts as a tumor promoter by stimulating acinar cell proliferation in rat pancreas. To this end, indices of pancreatic growth, including increased pancreatic weight, stimulation of acinar cell proliferation, and/or enhanced expression of immediate-early oncogenes were monitored in rats given gabapentin in the diet at 2 g/kg/day for up to 12 months. Rats fed raw soy flour (RSF), a known inducer of pancreatic acinar cell tumors through cholecystokinin-mediated mitogenic stimulation, were used throughout as positive controls. In addition, recent data suggests that gabapentin binds to the alpha(2)delta subunit of a voltage-gated, L-type calcium channel. Because signaling pathways for proliferative processes in pancreatic acinar cells involve intracellular calcium mobilization, the effects of gabapentin on intracellular calcium mobilization ([Ca(2+)](i)) and (3)H-thymidine incorporation were investigated in pancreatic acinar cells isolated from normal rat pancreas and in the AR42J rat pancreatic tumor cell line. As indicated by BrdU labeling indices, acinar cell proliferation increased 3-fold by Day 3 of RSF treatment and remained slightly greater than controls throughout the experiment. Pancreatic weights of RSF-fed rats were 32 to 56% greater than controls throughout the experiment. In contrast, gabapentin had no effect on pancreatic weight or acinar cell labeling index, and therefore had no apparent effect on pancreatic growth. In isolated pancreatic acinar cells, however, gabapentin induced mobilization of intracellular calcium and caused a slight increase in (3)H-thymidine incorporation. The data suggest that gabapentin may possess low level mitogenic activity, which is not easily detectable in in vivo assays. PMID- 10788560 TI - Apoptosis and P53 induction in human lung fibroblasts exposed to chromium (VI): effect of ascorbate and tocopherol. AB - Some forms of hexavalent chromium [Cr(VI)] are known to cause damage to respiratory tract tissue, and are thought to be human lung carcinogens. Because Cr(VI) is mutagenic and carcinogenic at doses that evoke cell toxicity, the objective of these experiments was to examine the effect of Cr(VI) on the growth, survival, and mode of cell death in normal human lung fibroblasts (HLF cells). DNA adduct formation was monitored as a marker for bioavailability of genotoxic chromium. We also examined the modulation of these endpoints by vitamins C and E. Long-term Cr(VI) exposures were employed, which decreased clonogenic cell survival by 25% to 95% in a dose-dependent manner. The predominant cellular response to Cr(VI) was growth arrest. We found that Cr(VI) caused up to 20% of HLF cells to undergo apoptosis, and documented apoptotic morphology and the phagocytosis of apoptotic bodies by neighboring cells. P53 levels increased 4- to 6-fold in chromium-treated cells. In contrast with previous studies using CHO cells, the present study using HLFs found that pretreatment with either vitamin C or E did not exhibit a significant effect on Cr-induced apoptosis or clonogenic survival. In addition, pretreatment with vitamin C did not affect the p53 induction observed after chromium treatment. Neither vitamin had any effect on Cr DNA adduct formation. These data indicate that although pretreatment with vitamin C or E alters the spectrum of cellular and/or genetic lesions induced by chromium(VI), neither vitamin altered the initiation or progression of apoptosis in diploid human lung cells. PMID- 10788561 TI - A rapid and sensitive reporter gene that uses green fluorescent protein expression to detect chemicals with estrogenic activity. AB - A reporter gene sequence was constructed within a eukaryotic expression vector. The altered plasmid contained 2 sequential estrogen response elements (ERE) coupled to a human phosphoglycerate kinase (PGK) promoter inserted upstream from a cDNA sequence encoding enhanced green fluorescent protein (GFP) with a 3' polyadenylation signal. The plasmid was linearized and transfected into MCF-7 cells, a human breast cancer-derived line that expresses the estrogen receptor (ER). No selectable marker was present in the plasmid, requiring stably transfected cells to be selected by fluorescence-activated cell sorting based on GFP expression after the cells were treated with 10(-9) M 17beta-estradiol (E2). Stably transfected MCF-7 cells (MCF7-ERE) exhibited 2000-3000 times more fluorescence at 488 nm excitation and 512 nm emission than non-transfected cells. MCF7-ERE cells exhibited a linear increase in GFP expression induced over a range of 10(-12) M E2, a concentration giving 2 times the background expression, to maximal expression at 3 x 0(-10) M E2. From the maximal level, GFP expression plateaued, and then declined when E2 was increased to the highest concentration tested, 10(-7) M. 4-Hydroxytamoxifen (TFN-OH) treatment of cells produced a dose dependent inhibition of E2-induced GFP expression, indicating the interaction of ER in the regulation of GFP gene expression. A series of estrogenic chemicals were evaluated for their capacity to induce GFP expression in MCF7-ERE cells, showing induced expression of GFP at concentrations 2-4 log units higher than the E2 concentration giving maximal GFP expression. The ERE-PGK-GFP reporter gene system is capable of rapid GFP expression in the presence of low concentrations of E2, and of quantifying estrogenicity of chemicals compared with a standard curve of the natural ligand, 17beta-estradiol. PMID- 10788563 TI - Effect of micelle fatty acid composition and 3,4,3', 4'-tetrachlorobiphenyl (TCB) exposure on intestinal [(14)C]-TCB bioavailability and biotransformation in channel catfish in situ preparations. AB - Polychlorinated biphenyls are transferred in the diet along aquatic food chains. This study investigated the effect of dietary micelle composition and 3,4,3',4' tetrachlorobiphenyl (TCB) exposure upon the subsequent systemic bioavailability and intestinal metabolism of [(14)C]-TCB in a catfish in situ intestinal preparation. Initial in vitro experiments examined the solubility of [(14)C]-TCB in micelles of varying fatty acid composition. Micelles composed of single fatty acids demonstrated greater [(14)C]-TCB solubility with those fatty acids of longer chain length. Similarly, micelles of the long-chain fatty acid, linoleic acid, solubilized more [(14)C]-TCB than mixed micelles formulated from equal amounts of myristic (14:0), palmitic (16:0), stearic (18:0), or linoleic (18:2) acids. Systemic bioavailability of [(14)C]-TCB (60 microM) from an in situ perfused intestinal preparation was 2.2-fold greater when delivered to the intestine in linoleic acid micelles as compared to the mixed micelle preparation. Catfish exposed in vivo to either 0.5 or 5.0 mg TCB/kg feed for 10 days resulted in a 45 to 47% decrease in the subsequent systemic bioavailability of [(14)C]-TCB in the in situ intestinal preparation. Total intestinal cytochrome P450 content was not significantly affected by TCB preexposure. Immunodetectable CYP1A was found only in the 5.0 mg TCB/kg diet treatment. Corresponding intestinal aryl hydrocarbon hydroxylase (AHH) activities were 2.46 +/- 1.16, 2.43 +/- 1.58, and 11.35 +/- 10.25 pmol/min/mg protein for the control, 0.5, and 5 mg TCB/kg diet groups, respectively. [(14)C]-TCB in the in situ preparation was metabolized to only a small degree upon a single pass through the intestinal mucosa of the catfish. High variability and low rates of metabolism precluded the association of the magnitude of metabolism with dietary TCB pretreatment. Analysis of tissue sample extracts demonstrated 4 minor peaks, 3 of which were tentatively identified by co-elution with standards as 2-OH-3,4,3',4'-TCB, 4-OH-3,5,3',4' TCB, and 5-OH-3, 4,3',4'-TCB. A fourth remains unidentified. Histological changes in the intestine such as thinning of the submucosa and increased numbers of goblet cells were evident at the 5.0 mg TCB/kg diet dose. These results suggest that TCB intestinal bioavailability may be linked to micelle composition as well as TCB exposure history. Furthermore, single pass intestinal metabolism appears to be a minor contributor to the biotransformational modification of dietary TCB. PMID- 10788562 TI - Differential effects of microsomal enzyme inducers on in vitro thyroxine (T(4)) and triiodothyronine (T(3)) glucuronidation. AB - Microsomal enzyme inducers that increase UDP-glucuronosyltransferase (UDP-GT) activity are suspected to affect the thyroid gland by increasing the glucuronidation of T(4), which reduces serum thyroxine (T(4)). In response to reduced serum T(4), serum thyroid-stimulating hormone (TSH) increases. However, not all microsomal enzyme inducers that reduce serum T(4) produce an increase in serum TSH. We have shown that serum TSH is increased the most in rats treated with the microsomal enzyme inducers phenobarbital (PB) or pregnenolone-16alpha carbonitrile (PCN), whereas TSH is affected less in rats treated with 3 methylcholanthrene (3MC) and Aroclor 1254 (PCB). It is unclear why serum TSH is differentially affected by various microsomal enzyme inducers. We propose that the glucuronidation of T(3) might be the reason serum TSH is increased by some microsomal enzyme inducers but not by others. Male Sprague-Dawley rats were fed either a basal diet or a diet containing PB (at 300, 600, 1200, or 2400 ppm), PCN (at 200, 400, 800, or 1600 ppm), 3MC (at 50, 100, 200, or 400 ppm), or PCB (at 25, 50, 100, or 200 ppm) for 7 days; and T(4) and T(3) UDP-GT activities were then determined. T(4) UDP-GT activity was increased in rats treated with PB (120%), PCN (250 to 400%), 3MC (400 to 600%), or PCB (300 to 430%). In contrast, T(3) UDP-GT activity was increased in rats treated with PB (90%) or PCN (120 to 200%), whereas 3MC and PCB treatments did not have an appreciable effect. In conclusion, differential effects on T(3) glucuronosyltransferase activity were found in rats treated with microsomal enzyme inducers. PMID- 10788564 TI - Low-dose whole-body irradiation (LD-WBI) changes protein expression of mouse thymocytes: effect of a LD-WBI-enhanced protein RIP10 on cell proliferation and spontaneous or radiation-induced thymocyte apoptosis. AB - Low-dose radiation (LDR) can potentiate cellular metabolic activities or immune functions in vivo (hormesis), and can render cells resistant to DNA or chromosome damage caused by subsequent high-dose radiation (adaptive response). Protein synthesis was required for these cellular responses to LDR. In the present study, the early expression of proteins by thymocytes in response to low-dose whole-body irradiation (LD-WBI) was investigated. The expression of novel and previously existing proteins was found in the nucleus, cytoplasm, and extracellular fluid of thymocytes at 4 hours after WBI with 75-mGy X-rays. A 10 kD protein (RIP10) was seen in the cytoplasm of thymocytes after LD-WBI was further investigated. The fraction containing RIP10 separated by Sephadex G 100 gel filtration potentiated spontaneous thymocyte, and mitogen-induced splenocyte proliferation. Western blotting demonstrated that an anti-RIP10 antibody could react with a 10-kD cytoplasm protein and also with a 13-kD nuclear protein in thymocytes at 4 h after LD-WBI. Immunocytochemical staining showed the existence of RIP10 in several immune tissues including thymus, spleen, and lymph node. RIP10 expression, as determined by immunocytochemical staining and flow cytometry, was enhanced at 4-8 h after LD-WBI. Cell-cycle arrest (G(0)/G(1) block with decreased percentage of S-phase cells), and increased levels of spontaneous or radiation induced apoptosis were observed in thymocytes incubated with RIP10 antibody in vitro for 4 h or 24 h. These results directly demonstrated the role of RIP10 in modulating cell proliferation and apoptosis. This finding is important to understand the mechanisms underlying LDR-induced hormesis and adaptive response. PMID- 10788565 TI - Ah receptor-based chemical screening bioassays: application and limitations for the detection of Ah receptor agonists. AB - The aromatic hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biologic and toxicologic effects of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related chemicals. Here we utilized two AhR-dependent bioassay systems as screening tools to identify novel AhR agonists and to detect the presence of AhR agonists in sample extracts. These assays measure the ability of a chemical to activate AhR DNA binding in vitro (GRAB bioassay) or AhR-dependent (luciferase) gene expression in cultured cells (CALUX bioassay). Known AhR agonists (halogenated and nonhalogenated aromatic hydrocarbons) were positive in both assays, whereas the AhR antagonist alpha naphthoflavone exhibited agonist activity only in the GRAB assay. In vitro GRAB analysis has identified several imidazoline receptor ligands and beta-carbolines as AhR agonists and also revealed the presence of AhR agonist activity in crude DMSO extracts of commercial newspapers. In contrast to their positive activity in the GRAB assay, the majority of these chemicals/extracts were only weakly active or inactive in the cell-based CALUX assay. Our results not only reveal that the ability of a chemical to activate the AhR in vitro does not necessarily correlate with its ability to induce gene expression in intact cells, but the high level of false positives obtained with the GRAB assay clearly demonstrates its inability to accurately identify AhR agonists or agonist activity. Screening of unknown chemicals, chemical classes, and samples for AhR agonist activity will require the use of intact cell bioassays. PMID- 10788566 TI - 1-Bromopropane, an alternative to ozone layer depleting solvents, is dose dependently neurotoxic to rats in long-term inhalation exposure. AB - 1-Bromopropane has been newly introduced as an alternative to ozone layer depleting solvents. We aimed to clarify the dose-dependent effects of 1 bromopropane on the nervous system. Forty-four Wistar male rats were randomly divided into 4 groups of 11 each. The groups were exposed to 200, 400, or 800 ppm of 1-bromopropane or only fresh air 8 h per day for 12 weeks. Grip strength of forelimbs and hind limbs, maximum motor nerve conduction velocity (MCV), and distal latency (DL) of the tail nerve were measured in 9 rats of each group every 4 weeks. The other 2 rats of each group were perfused at the end of the experiment for morphological examinations. The rats of the 800-ppm group showed poor kicking and were not able to stand still on the slope. After a 12-week exposure, forelimb grip strength decreased significantly at 800 ppm and hind limb grip strength decreased significantly at both 400 and 800 ppm or after a 12-week exposure. MCV and DL of the tail nerve deteriorated significantly at 800 ppm. Ovoid or bubble-like debris of myelin sheaths was prominent in the unraveled muscular branch of the posterior tibial nerve in the 800-ppm group. Swelling of preterminal axons in the gracile nucleus increased in a dose-dependent manner. Plasma creatine phosphokinase (CPK) decreased dose-dependently with significant changes at 400 and 800 ppm. 1-Bromopropane induced weakness in the muscle strength of rat limbs and deterioration of MCV and DL in a dose-dependent manner, with morphological changes in peripheral nerve and preterminal axon in the gracile nucleus. 1-Bromopropane may be seriously neurotoxic to humans and should thus be used carefully in the workplace. PMID- 10788567 TI - Comparative cholinergic neurotoxicity of oral chlorpyrifos exposures in preweanling and adult rats. AB - Chlorpyrifos (CPF) is a common organophosphorus (OP) pesticide. Previous studies have demonstrated that neonatal rats are more sensitive than adults to the acute toxicity of high dosages of CPF. The present study examined lethality and age related differences in neurochemical indicators and functional signs of neurotoxicity following a broad range of acute and repeated oral CPF exposures. There was about a 9-fold difference in sensitivity to the acute-dose lethality of chlorpyrifos among neonatal (7 days-of-age) and adult (90 days-of-age) rats (LD(10): neonates = 15 mg/kg; adults = 136 mg/kg), while juvenile rats (21 days of-age) exhibited intermediate sensitivity (LD(10) = 47 mg/kg). Neonatal and adult rats (n = 5-7/treatment/age group/time point) were given CPF (0, 0.15, 0.45, 0. 75, 1.5, 4.5, 7.5, or 15 mg/kg/day) for 14 days and sacrificed 4 h after either the first or 14th dose for neurochemical measurements (cholinesterase activity in frontal cortex, plasma and RBC, and muscarinic ([(3)H]QNB) and nicotinic ([(3)H]epibatidine) receptor binding in frontal cortex. No overt signs of functional toxicity (involuntary movements, SLUD signs) were noted in either age group by 4 h after the first dose. With repeated CPF exposures, however, signs of cholinergic toxicity were noted in both age groups at the higher dose levels [no observed effect levels (NOELs): neonate = 4.5 mg/kg/day; adult = 7.5 mg/kg/day]. Similar degrees of ChE inhibition were noted in neonatal brain and blood fractions following acute exposure, but substantial ChE inhibition was only noted in adult plasma and RBC 4 h after the first treatment. Following repeated CPF exposures, similar degrees of ChE inhibition were again noted in tissues from immature animals, but a wide range of sensitivity to inhibition was noted in adult tissues. NOELs based on ChE inhibition for adults were about 1->/=10-fold higher than in neonates with acute exposure but only 0.2-2 times higher with repeated dosing. Moreover, dose-related inhibition of brain ChE was similar between age groups, and similar reductions in both QNB and epibatidine binding were noted between the age groups after repeated dosing, even though by the end of the dosing period young animals (juveniles) were still about 3 times more sensitive than adults, based on acute lethality. We conclude that while immature animals can be markedly more sensitive to lethal effects of high doses of CPF, lesser or no age-related differences are apparent, based on non-lethal endpoints, in particular with repeated exposures. PMID- 10788568 TI - An evaluation of l-ephedrine neurotoxicity with respect to hyperthermia and caudate/putamen microdialysate levels of ephedrine, dopamine, serotonin, and glutamate. AB - l-Ephedrine is an active ingredient in several herbal formulations with a mechanism of action similar to amphetamine and methamphetamine. However, its potential to damage dopaminergic terminals in the caudate/putamen (CPu) has yet to be fully evaluated. The studies here used in vivo brain microdialysis experiments to determine the systemic doses and extracellular brain levels of l ephedrine necessary to produce similar increases in CPu extracellular dopamine and marked hyperthermia that were previously shown necessary for amphetamine induced neurotoxicity in male Sprague-Dawley rats. At an environmental temperature of 23 degrees C, a single 40 mg/kg intraperitoneal (ip) dose of l ephedrine produced marked hyperthermia (>/= 40 degrees C), peak microdialysate ephedrine levels of 7.3 +/- 1.2 microM, and a 20-fold increase in microdialysate dopamine levels. Twenty-five mg/kg produced a lesser degree of hyperthermia, peak microdialysate ephedrine levels of 2.6 +/- 0.4 microM, and a 10-fold increase in dopamine levels. Three doses of 40 mg/kg given at 3-h intervals or 4 doses of 25 mg/kg l-ephedrine given at 2-h intervals were compared with 4 doses of 5 mg/kg d amphetamine given at 2-h intervals. Multiple doses of either ephedrine or amphetamine caused severe hyperthermia (>/= 41.3 degrees C) but striatal tissue levels of dopamine 7 days after dosing were reduced only 25% or less by ephedrine compared to the 75% reductions produced by amphetamine. The increases in CPu microdialysate levels of serotonin produced by either 4 x 25 mg/kg l-ephedrine or 4 x 5 mg/kg d-amphetamine did not significantly differ, but elevation of dopamine levels by d-amphetamine were over 2-fold times the level caused by l-ephedrine. Microdialysate glutamate levels were elevated to the same extent by either 25 mg/kg l-ephedrine or 4 x 5 mg/kg d-amphetamine. l-Ephedrine may not be as neurotoxic to dopaminergic terminals as d-amphetamine, because non-lethal doses of l-ephedrine do not sufficiently increase the CPu dopamine levels within nerve terminals or the extracellular space to those necessary for a more pronounced long-term dopamine depletion. PMID- 10788569 TI - Dose-dependent alterations in androgen-regulated male reproductive development in rats exposed to Di(n-butyl) phthalate during late gestation. AB - Di(n-butyl) phthalate (DBP) is a commercially important plasticizer and ubiquitous environmental contaminant. Since previous, limited dose-response studies with DBP that reported alterations in male reproductive development and function failed to establish a NOAEL (no-observed-adverse-effect level), an extensive dose-response study was conducted. Pregnant CD rats were given DBP by gavage at 0, 0.5, 5, 50, or 100 mg/kg/day (n = 19-20) or 500 mg/kg/day (n = 11) from gestation day 12 to 21. In male offspring, anogenital distance was decreased at 500 mg DBP/kg/day. Retained areolas or nipples were present in 31 and 90% of male pups at 100 and 500 mg/kg/day, respectively. Preputial separation was not delayed by DBP treatment in males with normal external genitalia, but cleft penis (hypospadias) was observed in 5/58 rats (4/11 litters) at 500 mg/kg/day. Absent or partially developed epididymis (23/58 rats in 9/11 litters), vas deferens (16/58 animals in 9/11 litters), seminal vesicles (4/58 rats in 4/11 litters), and ventral prostate (1/58 animals) occurred at 500 mg/kg/day. In 110-day-old F(1) males, the weights of the testis, epididymis, dorsolateral and ventral prostates, seminal vesicles, and levator ani-bulbocavernosus muscle were decreased at 500 mg/kg/day. At 500 mg/kg/day, widespread seminiferous tubule degeneration was seen in 25/58 rats (in 9/11 litters), focal interstitial cell hyperplasia in 14/58 rats (in 5/11 litters), and interstitial cell adenoma in 1/58 rats (in 1/11 litters). For this 10-day prenatal (embryonic and fetal) exposure to DBP, the NOAEL and LOAEL (lowest-observed-adverse-effect level) were 50 and 100 mg/kg/day, respectively. This is currently the lowest NOAEL described for the toxicity of DBP. PMID- 10788570 TI - Characterization of the period of sensitivity of fetal male sexual development to vinclozolin. AB - Vinclozolin is a fungicide whose metabolites are androgen receptor (AR) antagonists. Previous work in our laboratory showed that perinatal administration of vinclozolin to rats results in malformations of the external genitalia, permanent nipples, reduced anogenital distance (AGD), and reduced seminal vesicle, ventral prostate, and epididymal weights. The objectives of this study were to determine the most sensitive period of fetal development to antiandrogenic effects of vinclozolin and to identify a dosing regime that would induce malformations in all of the male offspring. Pregnant rats were dosed with 400 mg vinclozolin/kg/day on either GD 12-13, GD 14-15, GD 16-17, GD 18-19, or GD 20-21, or with corn oil (2.5 ml/kg) from GD 12 through GD 21 (Experiment 1). All 2-day periods in which significant effects were produced were included in an extended dosing period, GD 14 through GD 19, in which pregnant rats were dosed with 200 or 400 mg vinclozolin/kg (Experiment 2). In Experiment 1, significant effects of vinclozolin were observed in rats dosed on gestation days (GD) 14-15, GD 16-17, and GD 18-19, while the most significant effects were observed in rats treated on GD 16-17. These effects include reduced AGD; presence of areolas, nipples, and malformations of the phallus; and reduced levator ani/bulbocavernosus weight. In contrast, ventral prostate weight was reduced only in the GD 18-19 group. The expanded dosing regime (Experiment 2) increased the percentage of male offspring with genital malformations (> 92%), and retained nipples (100%), further reduced the weight of the ventral prostate, and reduced the weight of the seminal vesicles. In addition, malformations were more severe and included vaginal pouch and ectopic/undescended testes. The latter was induced only in the 400 mg/kg group. These data indicate that the reproductive system of the fetal male rat is most sensitive to antiandrogenic effects of vinclozolin on GD 16 and 17, although effects are more severe and 100 % of male offspring are affected with administration of vinclozolin from GD 14 through GD 19. PMID- 10788571 TI - Relaxant effects of aflatoxins on isolated guinea pig trachea. AB - Dyspnea is one of the symptoms of acute aflatoxicosis. Contrary to expectations, we observed that naturally occurring aflatoxins (AF) AFB(1), AFB(2), AFG(1), and AFG(2) and their major metabolites AFM(1), AFM(2), AFP(1), AFQ(1), and AFG(2a) relaxed carbachol (C) precontracted guinea pig trachea to different degrees. The efficacies but not the potencies of AFB(1), AFB(2), AFG(1), and AFG(2) were similar to that of the beta-agonist, isoprenaline, whose activity was potentiated by the AF. Their mechanism of action is not clearly understood but several mechanistic indications were obtained with AFB(1): 1) its effect was not influenced by the beta-blocker, timolol, indicating that a direct interaction with beta(2)-adrenergic receptors was not involved. 2) AFB(1) potentiated PGE(1) and PGE(2), two relaxant prostaglandins, and its activity was reduced by indomethacin. 3) The cAMP level in the guinea pig trachea relaxed by AFB(1) increased, possibly due to inhibition of phosphodiesterase; direct interaction with PG receptors; and/or interaction with A(2) adenosinic receptors, suggested by the inhibitory activity of XAC, a specific antagonist. 4) Finally, since tetrodotoxin reduced the relaxant activity of AFB(1), it is speculated that this mycotoxin could stimulate inhibitory nonadrenergic, noncholinergic nerves (i NANC). In conclusion, the symptoms of acute aflatoxicosis do not seem to be due to a direct activity on the tracheal muscle, but rather, to the well-known pro inflammatory activity of the aflatoxins, which are capable of releasing arachidonic acid from cell membranes. PMID- 10788572 TI - Modeling normal aging bone loss, with consideration of bone loss in osteoporosis. AB - A physiologically based model of normal bone loss in human aging is presented. The model is a modification of an existing physiologically based model of body and bone growth from birth to maturity. To account for loss of bone after peak bone mass is reached between ages 25 and 30 years, a slow first-order loss of bone is incorporated into the existing model. The rate constants for this first order loss are the same for men and women but differ with the type of bone, being 3%/decade for cortical bone and 7-11%/decade for trabecular bone. In women, a 10 year period of more rapid loss of both cortical and trabecular bone is superimposed on the slow loss, beginning at the time of menopause. The superimposed loss occurs at the same relative rate in cortical and trabecular bone. Alterations in parameter values allow simulation of bone mass in osteoporotic men and women. The model is calibrated to quantitative estimates of cortical and trabecular bone mass as functions of age; in particular, to data sets of fractional vertebral bone volume as functions of age, and it is compared to the International Commission on Radiological Protection trend curves for skeletal mass in men and women to age 60. It is also applied to the question of whether loss of bone in women after menopause could create a hazard related to the return to blood of lead previously stored in bone. In agreement with observations made during 1976-1980, the model simulates an increase due to bone resorption of approximately 1 microg/dl in blood lead concentration in a postmenopausal (60-year-old) woman compared with a premenopausal (50-year-old) woman with typical lifetime ambient lead exposure. PMID- 10788573 TI - Ocular changes in beagle dogs following oral administration of CGS 24565, a potential hypolipidemic agent. AB - (11R)-N,15-dideoxo-1-deoxy-1,15-epoxy-11-hydroxy-4-0methy l-8-0-(2, 2-dimethyl-1 oxopropyl)-3-[4-?(2,4, 6-trimethylphenyl)methyl?-1-piperazinyl]rifamycin has been evaluated as a potential hypolipidemic agent. As part of a safety evaluation program, a 3-month oral toxicity study was performed in which CGS 24565 was administered to beagle dogs via gelatin capsules at 10, 50, or 300 mg/kg/day. Ophthalmoscopic examinations (using focal illumination and indirect opthalmoscopy) on day 83 (week 12) revealed bilateral adnexal and corneal changes affecting 5 dogs (3 males, 2 females, 300 mg/kg/day). Ophthalmoscopically, dogs from the 300 mg/kg dose level exhibited the adnexal changes characterized as ptosis, conjunctivitis, episcleritis, and relaxed membrane nictitans, while the corneal changes were characterized as posterior stromal edema (cloudy, diffuse opacity usually accompanied by deep neovascularization; the diffuse edema masked the complete evaluation of other ocular structures) and stromal infiltrates in the area of Decement's membrane (appeared to be multifocal, polymorphic changes/alterations in Decement's membrane, or endothelial swelling). No changes from normal were seen clinically in the eyes of other dogs on this experiment. In those dogs affected by the ocular changes caused by CGS 24565, a visual deficit in acuity was suspected. The corneal changes, as manifested, were suggestive of permanent, irreversible corneal damage. Subsequent ophthalmoscopic examinations performed at established intervals during weeks 15 through 26, revealed abatement of the adnexal changes, while the corneal changes, as described above, remained generally unchanged, confirming irreversibility of the corneal changes within the recovery period of 13 weeks. Light microscopy confirmed irreversible corneal neovascularization, vacuolar degeneration of the keratocytes at 300 mg/kg, and polymorphic infiltrates in the region of Decement's membrane. The results demonstrate that the cornea was the target tissue of toxicity for CGS 24565, and indicated that the findings represent a significant toxic effect. The correlation of histopathological findings support the hypothesis of the diagnosis of interstitial stromal degeneration/atrophy. The potential for a similar result to the cornea of humans does exist. Due to these changes and other toxic effects associated with this class of compound, further development was terminated. PMID- 10788574 TI - Effects of glutaraldehyde in a 2-year inhalation study in rats and mice. AB - Whole-body inhalation toxicology and carcinogenicity studies were performed with the widely used fixative and cold-sterilant glutaraldehyde. Groups of 50 male and female F344/N rats and B6C3F(1) mice were exposed to glutaraldehyde (rats: 0, 250, 500, or 750 ppb; mice: 0, 62.5, 125, or 250 ppb) 6 h/day, 5 days/week, for 104 weeks. Survival of 500- and 750-ppb female rats was less than that of controls. Mean body weights of all exposed groups of male rats, 500- and 750-ppb female rats, and 250-ppb female mice were generally less than those of controls. No exposure-related neoplastic lesions were observed in either rats or mice. Non neoplastic lesions were limited primarily to the most anterior region of the nasal cavity. In rats, hyperplasia and inflammation of the squamous epithelium; hyperplasia, goblet cell hyperplasia, inflammation, and squamous metaplasia of the respiratory epithelium; and hyaline degeneration of the olfactory epithelium were observed. In mice, the nasal lesions were qualitatively similar to those in rats. Squamous metaplasia of the respiratory epithelium was observed in both sexes of mice while female mice also had inflammation and hyaline degeneration of the respiratory epithelium. In contrast to the nasal carcinogen formaldehyde, no neoplastic lesions were observed after inhalation exposure to glutaraldehyde. However, exposure to glutaraldehyde resulted in considerable non-neoplastic lesions in the noses of rats and mice. PMID- 10788575 TI - Diethyldithiocarbamate enhances production of nitric oxide and TNF-alpha by lipopolysaccharide-stimulated rat Kupffer cells. AB - Previous studies have shown that large doses of diethyldithiocarbamate (DDC) cause liver injury in rats and the pathogenesis of this injury involves, in part, release of superoxide anion by Kupffer cells. The purpose of this study was to evaluate if DDC was able to stimulate other potentially toxic mediators such as nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) using isolated rat Kupffer cells. DDC alone did not stimulate the release of NO and TNF-alpha by Kupffer cells. Interestingly, when Kupffer cells were stimulated by lipopolysaccharide (LPS), DDC (0-30 microM) enhanced the production of both NO and TNF-alpha in a concentration-dependent manner. Therefore, we further studied how DDC modulated the response of Kupffer cells to LPS. Immunocytochemical studies revealed that DDC increased the amount of inducible NO synthase and TNF alpha protein in Kupffer cells after their exposure to LPS. The enhanced effects of DDC on the release of NO and TNF-alpha from Kupffer cells was inhibited by N acetyl-L-cysteine (an inhibitor of transcription factor NF-kappaB activation). By using a specific antibody for NF-kappaBp65, it was found that DDC enhanced the LPS-activated nuclear translocation of NF-kappaB. There was no evidence of intracellular oxidative stress following either LPS alone or DDC + LPS exposure. The stimulatory effect of DDC on both NO and TNF-alpha release was inhibited by H 7 (an inhibitor of protein kinase C) but not H-8 (an inhibitor of cAMP-dependent protein kinase). These findings demonstrate that DDC enhances the production of NO and TNF-alpha by LPS-stimulated Kupffer cells and suggest that protein kinase C plays a critical role in mediating these effects of DDC. PMID- 10788576 TI - A robust structure-activity relationship (SAR) model for esters that cause skin irritation in humans. AB - A structure-activity relationship (SAR) model has been developed to discriminate skin irritant from nonirritant esters. The model is based on the physicochemical properties of 42 esters that were tested in humans for skin irritation. Nineteen physicochemical parameters that represent transport, electronic, and steric properties were calculated for each chemical. Best subsets regression analysis indicated candidate models for further analysis. Regression analyses identified significant models (p < 0.05) that had variables that were also significant (p < 0.05). These candidate models were evaluated using linear discriminant analysis to determine if the irritant esters could be discriminated from nonirritant esters. The stability of the model was evident from the consistency of parameters among ten submodels generated using multiple random sampling of the database. The sensitivity of the ten models, evaluated by "leave-one-out" cross-validation, ranged from 0. 846 to 0.923, with a mean of 0.885 +/- 0.025 (95% CI). The specificity ranged from 0.615 to 0.923, with a mean of 0.738 +/- 0.06 (CI). Compared with nonirritant esters, irritant esters had lower density, lower water solubility, lower sum of partial positive charges, higher Hansen hydrogen bonding parameter, and higher Hansen dispersion parameter. The results indicate that physicochemical features of esters contribute to their ability to cause skin irritation in humans, and that chemical partitioning into the epidermis and intermolecular reactions are likely important components of the response. This model is applicable for prediction of human irritation of esters yet untested. PMID- 10788577 TI - Metallothionein-null mice are more sensitive than wild-type mice to liver injury induced by repeated exposure to cadmium. AB - Liver is a major target organ of cadmium (Cd) toxicity following acute and chronic exposure. Metallothionein (MT), a low-molecular-weight, cysteine-rich, metal-binding protein has been shown to play an important role in protection against acute Cd-induced liver injury. This study investigates the role of MT in liver injury induced by repeated exposure to Cd. Wild-type and MT-I/II knockout (MT I/II-null) mice were injected sc with a wide range of CdCl(2) doses, 6 times/week, for up to 10 weeks, and their hepatic Cd content, hepatic MT concentration, and liver injury were examined. Repeated administration of CdCl(2) produced acute and nonspecific chronic inflammation in the parenchyma and portal tracts and around central veins. Higher doses produced granulomatous inflammation and proliferating nodules in liver parenchyma. Apoptosis and mitosis occurred concomitantly in liver following repeated Cd exposure, whereas necrosis was mild. As a result, significant elevation of serum enzyme levels was not observed. In wild-type mice, hepatic Cd concentration increased in a dose- and time-dependent manner, reaching 400 microgram/g liver, along with 150-fold increases in hepatic MT concentrations, the latter reaching 1200 microgram/g liver. In contrast, in MT I/II-null mice, hepatic Cd concentrations were about 10 microgram/g liver. Despite the lower accumulation of Cd in livers of MT I/II-null mice, the maximum tolerated dose of Cd was one-eighth lower than that for wild-type mice at 10 weeks, and liver injury was more pronounced in the MT I/II-null mice, as evidenced by increases in liver/body weight ratios and histopathological analyses. In conclusion, these data indicate that (1) nonspecific chronic inflammation, granulomatous inflammation, apoptosis, liver cell regeneration, and presumably, preneoplastic proliferating nodules are major features of liver injury induced by repeated Cd exposure, and (2) intracellular MT is an important protein protecting against this Cd-induced liver injury. PMID- 10788578 TI - Polymer-supported reagents and catalysts: recent advances in synthetic applications. AB - The current surge in parallel array synthesis for the production of small molecule libraries has generated keen interest in the application of solid supported reagents and catalysts in solution-phase chemistry. The strategy assimilates the advantages of product isolation and purification of solid-phase organic synthesis with the flexible choice of chemistry from the vast repertoire of solution-phase organic reactions. This review summarizes the significant recent advances in the application of polymer-bound reagents and catalysts in solution-phase synthesis of organic molecules. Multi-step reaction sequences employing sequential use of polymer-supported reagents are also discussed. In view of the earlier review publications on this topic, only the recent literature covering 1998 and 1999 is included. PMID- 10788579 TI - Organic synthesis using chemical tags: the third Leg of parallel synthesis. AB - Increasing emphasis has recently been placed on the development of synthetic methods which effectively couple chemical synthesis and purification. For example, new formats for parallel synthesis are being developed which involve attachment of chemical tags to both reagents, reactants, and substrates to permit their chemoselective removal from reaction mixtures. The driving force for the development of tagged organic reagents is the ability to use standard solution phase chemistry methods and reaction monitoring techniques (e.g. TLC and HPLC). In this mini-review, we will outline recent developments on the growing class of chemically tagged reagents, reactants, and substrates and highlight examples of their use in multistep synthesis. PMID- 10788581 TI - Functionalized porous silica microspheres as scavengers in parallel synthesis. AB - The use of solid scavengers in parallel solution-phase organic synthesis is an effective method for work-up and purification. Functionalized macroreticular or gel-form polystyrene particles are generally used for scavenging applications, how ever these materials have some limitations. We have developed new scavenging reagents based on ultrapure silica microspheres displaying a variety of functional groups useful for sequestering impurities from reaction products. These materials are easy to handle, have excellent mass-transfer properties, and are efficient scavengers in both polar and nonpolar organic solvents. The properties of these materials were tailored specifically to fit the needs of a medicinal chemist employing parallel synthesis techniques in current commercial equipment. Results are presented from head-to-head comparisons with conventional scavengers in tests designed to demonstrate the versatility of these new materials. PMID- 10788580 TI - The use of polymer-bound triphenylphosphine in the stereochemical inversion of secondary alcohols. AB - Polymer-bound triphenylphosphine can replace triphenylphosphine in the Mitsunobu reaction to generate stereochemically inverted secondary alcohols. This method is comparable with the standard Mitsunobu reaction in terms of inversion of stereochemistry, yield, and reaction time, even for sterically very hindered secondary alcohols. The special merit of this reaction is that the excess polymer bound triphenylphosphine and its by-products are easily removed by filtration from the reaction products. PMID- 10788582 TI - Titanium(IV) isopropoxide mediated solution phase reductive amination on an automated platform: application in the generation of urea and amide libraries. AB - Amine libraries and their derivatives are important targets for high throughput synthesis because of their versatility as medicinal agents and agrochemicals. As a part of our efforts towards automated chemical library synthesis, a titanium(IV) isopropoxide mediated solution phase reductive amination protocol was successfully translated to automation on the Trident(TM) library synthesizer of Argonaut Technologies. An array of 24 secondary amines was prepared in high yield and purity from 4 primary amines and 6 carbonyl compounds. These secondary amines were further utilized in a split synthesis to generate libraries of ureas, amides and sulfonamides in solution phase on the Trident(TM). The automated runs included 192 reactions to synthesize 96 ureas in duplicate and 96 reactions to synthesize 48 amides and 48 sulfonamides. A number of polymer-assisted solution phase protocols were employed for parallel work-up and purification of the products in each step. PMID- 10788583 TI - Chemoselective high-throughput purification mediated by solid-supported reagents: its application to the first 6, 9-disubstituted purine library synthesis. AB - A new application of solid-supported reagents was developed to separate the alkylated N7/N9 regioisomers derived from commercially available 2-amino-6 chloropurine. Simple filtration through an alumina/H+ pad or scavenging by AG/Dowex-50W-X8 resin provides diverse N9 regioisomers selectively in moderate yields with high purities (>90%). This purification method can be conveniently used in a high-throughput format and facilitates the synthesis of a purine library without laborious regioisomer separation and aqueous work-up. The first library synthesis of 6,9-disubstituted purines is reported using the combination of this novel separation method in conjunction with polymer-supported reagents. PMID- 10788584 TI - Chromatography-free synthesis of 1,2,4-oxadiazoles using ROMPGEL-supported acylating reagents. AB - 1,2,4-Oxadiazoles were synthesized in solution from aromatic amidoximes and acylating agents supported on a ring opening metathesis polymer (ROMPGEL) backbone. High yields and purities of the 1,2,4-oxadiazoles were obtained with minimal purification. PMID- 10788585 TI - Multi-step polymer-assisted solution-phase (PASP) library synthesis of functionalized diaminobenzamides. AB - A parallel solution-phase library synthesis of functionalized diaminobenzamides is described. The four-step library synthesis is accomplished using polymer assisted solution-phase (PASP) synthesis techniques. This high-yielding, multi step sequence utilizes sequestering resins for the removal of reactants, reactant by-products, and employs a resin capture/release strategy as a key library synthesis step. Step one of the sequence relies on the displacement of an activated fluoro-group from the aromatic ring of 1a, b with a variety of primary amines to introduce the first diversity position. Step two is hydrolysis of the benzoate ester to a benzoic acid which is subsequently captured on a polyamine resin, washed, and released to give 4a, b in pure form. Step three utilizes PASP resins to mediate the amide coupling of a benzoic acid with a variety of primary amines to give the aminonitrobenzamides 5a, b and introduces the second diversity position. Step four is the parallel reduction of the aminonitrobenzamides 5a, b to the functionalized diaminobenzamides 6a, b. This library synthesis proceeds with high overall purities which average 80 % over the 4-step sequence. PMID- 10788586 TI - Fluorous-phase soluble polymeric supports. AB - Fluorous phase soluble polymer supports derived from fluoroacrylate polymers are described. N-Acryloxysuccinimide-containing fluoroacrylate polymers were readily prepared from commercially available monomers. The activated acrylates so prepared were then converted into chelating and non-chelating ligands by amidation of the N-acryloxysuccinimide active ester residues. Phosphine ligands attached to these supports were used to prepare neutral and cationic rhodium(I) hydrogenation catalysts as well as palladium(0) catalysts. Similar substitution of pendant active ester groups to form hydroxamic acid ligands for metal sequestration is also feasible. Liquid/liquid extraction readily separated, recycled and reused these polymer-bound ligands and catalysts. While fluorous phase solubility could be attained with polymers containing only heptafluorobutyryl groups, selective solubility in a fluorous phase in contact with an organic phase was only seen with fluoroacrylates that contained larger fluorinated ester groups. PMID- 10788587 TI - Signal transduction pathway targets for anticancer drug discovery. AB - There are currently over 80 agents officially approved for the treatment of cancer world-wide. However, the most common epithelial cancers, which cause greater than 75% of cancer deaths, remain incurable. Most drugs have been developed empirically by testing large numbers of chemicals on rapidly growing transplantable rodent tumors, and more recently, human tumor xenografts. This approach has identified prodeminantly DNA-active drugs that are considerably toxic and have limited efficacy. Novel molecular targets, which are selective for neoplastic cells, are needed for chemotherapeutic agents to improve cure rates of epithelial malignancies, with acceptable toxicity. In recent years, agents inhibiting signal transduction pathway molecules have entered clinical trials. These include antibodies and small molecules, which inhibit growth factor receptors and their receptor tyrosine kinases, inhibitors of cytoplasmic second messengers such as ras, raf and MEK, inhibitors of protein trafficking, and inhibitors of protein degradation. PMID- 10788588 TI - Cell cycle molecular targets in novel anticancer drug discovery. AB - A number of potential molecular targets for novel anticancer drug discovery have been identified in cell cycle control mechanisms. Prominent among these are the regulatory proteins, cyclins and their effector counterparts the cyclin dependent kinases (CDKs). Aberrant expression of these proteins, particularly cyclins involved in the G1 phase of the cell cycle, namely the D and E cyclins, has been associated with a variety of human cancers, including breast and colorectal cancer, B-lymphoma, prostate and non-small cell lung cancer. Inhibition of CDK kinase activity has turned out to be the most productive strategy for the discovery and design novel anticancer agents specifically targeting the cell cycle. Other potentially useful cell cycle areas for exploration include cyclin CDK interactions, Cdc25 activation of cyclin-CDK complexes, ubiquitin-mediated proteolysis of cyclins, cell cycle check point kinases like Chk1, and recently identified oncogenic cell cycle-related aurora and polo-like kinases. Potent specific inhibitors have been identified that bind to the ATP site of CDKs, mainly cyclin B-CDK1, cyclin A-CDK2, and cyclin D-CDK4 complexes, and inhibit kinase activity. X-ray crystallographic data of CDKs, and their complexes with inhibitors have played a major role in the success of drug discovery efforts. Combinatorial chemistry, highthroughput screening, functional genomics and informatics have also contributed. CDK inhibitors currently under investigation include flavopiridol, olomoucine, roscovitine, puvalanol B, the dihydroindolo[3,2 d][1]benzazepinone kenpaullone, indirubin-3 -monoxime and novel diaminothiazoles such as AG12275. The anticancer therapeutic potential of CDK inhibitors has been demonstrated in preclinical studies, and Phases I and II clinical trials in cancer patients are currently underway. PMID- 10788589 TI - MDM2 oncogene as a novel target for human cancer therapy. AB - The MDM2 oncogene was first cloned as an amplified gene on a murine double-minute chromosome in the 3T3DM cell line, a spontaneously transformed derivative of BALB/c 3T3 cells. The MDM2 oncogene has now been shown to be amplified or overexpressed in many human cancers. It also has been suggested that MDM2 levels are associated with poor prognosis of several human cancers. The most exciting finding is the MDM2-p53 autoregulatory feedback loop that regulates the function of the p53 tumor suppressor gene. The MDM2 gene is a target for direct transcriptional activation by p53, and the MDM2 protein is a negative regulator of p53. The MDM2 oncoprotein binds to the p53 protein, inhibiting p53 functions as a transcription factor and inducing p53 degradation. The p53 tumor suppressor has an important role in cancer therapy, with p53-mediated cell growth arrest and/or apoptosis being major mechanisms of action for many clinically used cancer chemotherapeutic agents and radiation therapy. Therefore, the MDM2-p53 interaction may be a target for cancer therapy. In addition, the negative regulation of p53 by MDM2 may limit the magnitude of p53 activation by DNA damaging agents, thereby limiting their therapeutic effectiveness. If the MDM2 feed-back inhibition of p53 is interrupted, a significant increase in functional p53 levels will increase p53-mediated therapeutic effectiveness. Several approaches have now been tested using this strategy, including polypeptides targeted to MDM2-p53 binding domain and antisense oligonucleotides that specifically inhibit MDM2 expression. In addition to the interaction with p53, the MDM2 protein has been found to have interactions with other cellular proteins such as pRb and E2F-1. Although the exact function and significance of these interactions are not fully understood, the p53-independent functions of MDM2 may have a role in cancer etiology and progression, indicating that the MDM2 oncogene is a potential molecular target for cancer therapy. PMID- 10788590 TI - Angiogenesis: new targets for the development of anticancer chemotherapies. AB - Angiogenesis is the process by which new blood vessels are formed from preexisting microvasculature. To ensure an adequate blood supply, tumor cells release angiogenic factors that are capable of promoting nearby blood vessels to extend vascular branches to the tumor. In addition, larger tumors have been shown to release angiogeneic inhibitory factors that prevent blood vessels from sending branches to smaller, more distant tumors that compete for oxygen and nutrients. Angiogenesis is a complex multistep biochemical process, and offers several potential molecular targets for non-cytotoxic anticancer therapies. Strategies for exploiting tumor angiogenesis for novel cancer drug discovery include: (i) inhibition of proteolytic enzymes that breakdown the extracellular matrix surrounding existing capillaries; (ii) inhibition of endothelial cell migration; (iii) inhibition of endothelial cell proliferation; (iv) enhancement of tumor endothelial cell apoptosis. There is also a host of miscellaneous agents that inhibit angiogenesis for which the specific mechanisms are not clear. Several methods have been developed for measuring antiangiogenic activity both in vitro and in vivo. Although there has been intensive research efforts focused at the phenomena of angiogenesis, as well as the search for antiangiogenic agents for more than two decades, many questions remain unanswered with regard to the overall biochemical mechanisms of the angiogenesis process and the potential therapeutic utility of angiogenic inhibitors. Nevertheless potent angiogenic inhibitors capable of blocking tumor growth have been discovered, and appear to have potential for development into novel anticancer therapeutics. However there are still hurdles to be overcome before these inhibitors become mainstream therapies. PMID- 10788591 TI - G-Quadruplex DNA as a target for drug design. AB - Telomeres are structures on the ends of chromosomes that are required for chromosomal stability. Telomeric DNA contains a single-stranded G-rich DNA overhang, which may adopt a G-quadruplex structure. Telomere shortening has been implicated in cellular senescence. Telomerase is an enzyme which synthesizes the G-rich strand of telomere DNA. Telomerase activity is highly correlated with cancer and may allow cancer cells to escape senescence. Based on these observations, telomerase has been proposed as a potential target for anticancer drug design. The targeting of telomerase is associated with potential problems, including the existence in some cancer cells of telomerase-independent mechanisms for telomere maintenance, and the long delay time between telomerase inhibition and effects on proliferation. One promising approach for inhibiting telomerase involves targeting the G-quadruplex DNA structures thought to be involved in telomere and telomerase function. Compounds that specifically bind G-quadruplex DNA may interact directly with telomeres, in addition to inhibiting telomerase, and produce more immediate antiproliferative effects. The diamidoanthraquinones, porphyrins, and perylene diimides have all been shown to bind G-quadruplex DNA and inhibit telomerase. Most of these compounds also bind double-stranded DNA and are cytotoxic at the concentrations required to inhibit telomerase; however, certain perylene diimides appear to be non-cytotoxic, G-quadruplex selective telomerase inhibitors. Biological characterization of such compounds may provide validation for the concept of the G-quadruplex as a target in drug design. PMID- 10788592 TI - Molecular targets in cancer drug discovery: cell-based profiling. AB - The phrase "molecular target-based drug discovery" usually implies an in vitro biochemical assay or battery of assays. One portion of the U.S. National Cancer Institute's drug discovery program, to the contrary, examines molecular targets for cancer therapy in a cell-based format. That approach has a number of significant limitations, but it has produced databases of significant utility on the activities and structures of tested compounds, as well as on molecular characteristics of the cell types used for testing. PMID- 10788593 TI - Emerging roles of carbohydrates and glycomimetics in anticancer drug design. AB - Tumorigenesis is accompanied by marked changes in the expression and presentation of various macromolecules at the cell surface. These tumor-associated adjustments result from the differential expression of genes coding for the production or post-translational modifications of these macromolecules during transformation to a particular tumor phenotype. In turn, tumor cells acquire distinct biophysical properties which set them apart from their normal counterparts. Alterations of carbohydrate structures and their organization on the surface of neoplastic cells is a hallmark of the tumorigenic and, most notably, the metastatic phenotype. Carbohydrate-protein and carbohydrate-carbohydrate interactions are critical events in the progression, dissemination and invasion of cancer cells. Many cell cell contacts and subsequent remodeling of the tumor microenvironment are mediated by cell-surface glycans. The discovery of agents that modulate these interactions or interfere with the processing of tumor associated oligosaccharides is a fervent area of research today. This review will highlight the current status of the use of carbohydrate-based compounds that are being evaluated as potential anticancer therapeutics. In addition, the use of structures based on glycopeptides and carbohydrate mimetics will also be discussed. PMID- 10788594 TI - Design and evaluation of hepatitis B virus inhibitors. AB - Antiviral therapy of chronic hepatitis B remains a major clinical problem worldwide. The design of new nucleoside analogs that inhibit hepatitis B virus (HBV) replication allowed their evaluation in in vitro and in vivo experimental models of HBV infection. This research has led to the discovery of the anti-HBV activity of lamivudine and its approval for the therapy of chronic hepatitis B. However, due to the development of viral resistance, strategies based on the combination of new inhibitors of HBV replication with immune modulatory approaches are urgently required. PMID- 10788595 TI - Novel anti-RSV dianionic dendrimer-like compounds: design, synthesis and biological evaluation. AB - Human Respiratory Syncytial Virus (RSV) is considered to be the leading cause of lower respiratory tract disease in infants and young children. RSV is also a common pathogen in immunocompromised adults and in the elderly. RSV infection can be epidemic and is evident worldwide. Ribavirin, a small molecule agent, and Synagis, a monoclonal neutralizing antibody, are the only approved drugs for treatment and prevention of RSV in high-risk patients. This review is focused on a group of novel and specific inhibitors discovered at Wyeth-Ayerst Research. Some of these inhibitors have IC50 <50 nM and are active against all the tested group A and B viruses. They also have shown good efficacy in cotton rats and primates. Mechanism of action studies indicate that the compounds inhibit the next step in infection after adsorption suggesting that fusion is the target. A strong relationship between the inhibitor structures and their anti-RSV activity was established. This relationship appears to derive from a multivalent interaction between the functional groupings of the inhibitors and the F protein, which seem to be highly complementary and directional. PMID- 10788596 TI - An analysis of the catalytic cycle of HIV-1 reverse transcriptase: opportunities for chemotherapeutic intervention based on enzyme inhibition. AB - This review describes each of the steps in the HIV-1 reverse transcriptase catalytic cycle and evaluates each of these steps as a potential point of inhibition of the enzyme and consequently viral replication. To date, two classes of approved drugs act on the reverse transcriptase. They are: (1) the nucleoside reverse transcriptase inhibitors which either directly inhibit the enzyme or serve as alternative substrates for catalysis (resulting in chain termination) and (2) the non-nucleoside reverse transcriptase inhibitors which bind to an allosteric site and adversely affect the function of the enzyme by slowing the rate of chemical catalysis. In order to provide the best possible analysis of the potential of each of the steps in the catalytic cycle as a site of inhibition, the molecular forces which determine the intrinsic binding affinities and specificity of natural components of the catalytic complex will be described in as much detail as possible. PMID- 10788597 TI - Anti-HIV-1 integrase drugs: how far from the shelf? AB - Chemotherapy of HIV-1 infection/AIDS currently employs inhibitors of two products of the viral pol gene, the reverse transcriptase and protease enzymes. However, a third product of the pol gene is essential for retroviral multiplication, the integrase. As no cellular homologue of HIV integrase has been described, potential inhibitors could be relatively nontoxic. Development of HIV-1 integrase inhibitors could have favorable implication for combination therapy, including potential synergy with currently available inhibitors, as well as prevention of the chronic carrier state and the emergence of resistant mutants. Although several classes of putative integrase inhibitors that been described, still no clinically useful anti-integration drugs are available. It is the structural and functional complexity of the integration process together with the limitations of the available in vitro assays that has made it problematic to develop inhibitors of the HIV integrase. In this review we summarize current knowledge concerning the biology of this enzyme and of the integration process, and discuss major classes representatives of integrase inhibitors considering the obstacles to the development of true anti-integrase drugs. PMID- 10788598 TI - Design of anti-HIV compounds: from nucleoside to nucleoside 5'-triphosphate analogs. Problems and perspectives. AB - To date, human immunodeficiency virus infection remains incurable although a variety of antiviral agents have been identified and characterized. Even though nucleoside analogs have been the most successful prodrugs, there remains the need to develop new compounds that exhibit a more favorable toxicity profile, less susceptible to cross-resistance, and greater efficacy. As prodrugs, the nucleoside analogs should be sequentially phosphorylated by cellular kinases to yield triphosphate form before they can inhibit HIV replication at the reverse transcriptase level. The efficiency of phosphorylation of nucleoside analogs is a key factor in their antiviral activity and strongly depends on nucleoside structure and cell type. In recent years, several attempts have been made to improve therapeutic potential of nucleoside analogs by the use of nucleotide prodrugs (pronucleotides), that can avoid the first step of phosphorylation. This review focuses on problems of intracellular phosphorylation of nucleoside analogs and perspectives of developing of a new class of nucleotide analogs modified at phosphate group as a form for the delivery of nucleotide analogs into the cell. PMID- 10788600 TI - Gene vectors for cytokine expression in vivo. AB - The understanding of cytokine networks and the exploitation of these networks for the treatment of immune and inflammatory diseases as well as cancer depend on in vivo delivery of cytokines. Due to instability of recombinant cytokine proteins, investigators have employed cytokine-encoding gene therapy vectors to induce high levels of cytokine expression in vivo. Numerous gene therapy vectors have been developed recently which are suitable for this purpose. Recent advances in the design of adenovirus, adeno-associated virus, poxvirus, retrovirus, lentivirus, and nonviral vectors are described here. Properties of the various vector systems which determine their usefulness for cytokine gene delivery are compared. The implementations of cytokine-encoding gene therapy vectors for analyzing immune responses and for the therapy of inflammatory disorders, immune disease, infections and cancer are reviewed. PMID- 10788599 TI - Consideration of cytokines as therapeutics agents or targets. AB - There has been an explosion of our knowledge in cytokine biology in the last decade. Such knowledge is being quickly translated into the identification of etiologies and improved prophylaxis and therapy of disease. While cytokines have the potential to be used as therapeutics or immune adjuvants for certain diseases, they may also be culprits as therapeutic targets in other diseases. This review article serves, as an introduction to the other five articles in this thematic issue each of which has a specific focus on the frontier of cytokine therapeutic biology. This review contains sections dealing with general cytokine properties, cytokine classifications, human conditions caused by cytokine under expression and over-expression, Th1 and Th2 paradigm, cytokine therapy for acute/chronic inflammatory conditions, cytokine therapy for infectious diseases, and cytokine therapy for cancer. PMID- 10788601 TI - Anti-inflammatory cytokines and cytokine antagonists. AB - Cytokines are critical mediators of protective host responses, including defense against microbial invasion and tumorigenesis. However, the production of specific proinflammatory cytokines must be tightly regulated and compartmentalized to prevent the overzealous expression of these molecules that can culminate in unabated inflammation and tissue injury. Cytokine production and/or biologic effects can be inhibited by a variety of endogenous molecules, including anti inflammatory cytokines, soluble cytokine receptors, and receptor antagonist proteins. Additionally, synthetic molecules have been constructed to selectively block the synthesis, post-translation processing, or receptor binding of proinflammatory cytokines. Relevant anti-inflammatory cytokines and cytokine inhibitors (both endogenous and synthetic) will be the subject of this review, with a particular emphasis on those anti-inflammatory cytokines and cytokine inhibitors that have been used experimentally or clinically in the treatment of diseases that are believed to be mediated by excessive proinflammatory cytokine responses. PMID- 10788602 TI - Therapeutic use of chemokines. AB - Chemokines are involved in a number of pathological processes, and therefore represent important targets. However, it has also become apparent that chemokines have exciting therapeutic applications in inflammatory, infectious and cancer related diseases. The following review will highlight the application of novel therapies including viral-encoded, recombinant, and genetically engineered chemokines to a number of diseases or disorders. Advances in the application of novel chemokine delivery procedures both at the research bench and the clinical bedside will also be discussed. Overall, the utilization of chemokines to prevent and treat disease has tremendous potential. PMID- 10788603 TI - Cytokine therapeutics for infectious diseases. AB - Cytokines are potent molecules, which function as growth factors and orchestrate both innate and adaptive immune responses. Over the last two decades the number of molecules in this class have greatly expanded, and as the biology of these factors is better understood, several of these factors have entered the clinical arena to support or augment components of the immune response. Recently the use of cytokines/growth factors has been studied in patients without a defective immune system but either have significant infection or infection with drug resistant organisms. The use of cytokines as adjuvants in the treatment of infectious diseases is reviewed both in the context of protein and gene-based therapies. PMID- 10788604 TI - Cytokine therapy for cancer. AB - Modulation of immune responses by the use of recombinant cytokines or cytokine genes is one of the strategies for cancer therapy. Although host immune responses are complex and many kinds of cells are involved, crucial steps for enhancing anti-tumor responses can be induced by a single or a few cytokines administered. However, cytokines may induce toxic reactions or produce no substantial effects, when the concentration is inappropriate. Administration of recombinant cytokine(s) has advantages in controlling the blood concentration and the biological activity that can be induced by the cytokine. Since cytokines are relatively unstable in vivo, cancer patients have to receive a large amount of the recombinant protein to maintain the required blood concentration for biological activity. Administration of the protein is thereby often toxic to the patients. In contrast, secretion of the cytokine from tumor or vehicle cells by gene transfer is another therapeutic maneuver. Previous preclinical studies have shown that cytokines which facilitate type 1 helper T (Th1) cells-mediated immune reactions but not Th2 cells-mediated reactions, when produced in tumors, are effective for anti-tumor responses. Several technical problems to express sufficient amounts of cytokines in appropriate target cells remain unresolved but the potential of cytokine gene therapy is being explored. Cytokine therapy trials also contributes to our present knowledge of how anti-tumor responses can be effectively produced in cancer patients, shedding the light on the generation of tumor-specific immunity in the patients. PMID- 10788605 TI - Active site residue 297 of Aspergillus niger phytase critically affects the catalytic properties. AB - The wild-type phytases from the Aspergillus niger strains NRRL 3135 and T213 display a three-fold difference in specific activity (103 versus 32 U/mg protein), despite only 12 amino acid differences that are distributed all over the sequence of the protein. Of the 12 divergent positions, three are located in or close to the substrate binding site. Site-directed mutagenesis of these residues in A. niger T213 phytase showed that the R297Q mutation (R in T213, Q in NRRL 3135) fully accounts for the differences in catalytic properties observed. Molecular modelling revealed that R297 may directly interact with a phosphate group of phytic acid. The fact that this presumed ionic interaction - causing stronger binding of substrates and products - correlates with a lower specific activity indicates that product (myo-inositol pentakisphosphate) release is the rate-limiting step of the reaction. PMID- 10788607 TI - Anomalous RNA substrates for mammalian tRNA 3' processing endoribonuclease. AB - Mammalian tRNA 3' processing endoribonuclease (3' tRNase) is an enzyme responsible for the removal of a 3' trailer from pre-tRNA. The enzyme can also recognize and cleave any target RNA that forms a pre-tRNA-like complex with another RNA. To investigate the interaction between 3' tRNase and substrates, we tested various anomalous pre-tRNA-like complexes for cleavage by pig 3' tRNase. We examined how base mismatches in the acceptor stem affect 3' tRNase cleavage of RNA complexes, and found that even one base mismatch in the acceptor stem drastically reduces the cleavage efficiency. Mammalian 3' tRNase was able to recognize complexes between target RNAs and 5'-half tDNAs, and cleave the target RNAs, although inefficiently, whereas the enzyme had no activity to cleave phosphodiester bonds of DNA. A relatively long RNA target, the Escherichia coli chloramphenicol acetyltransferase (CAT) mRNA, was cleaved by 3' tRNase in the presence of appropriate 5'-half tRNAs. We also demonstrated that an RNA complex of lin-4 and lin-14 from Caenorhabditis elegans can be recognized and cleaved by pig 3' tRNase. PMID- 10788606 TI - Long-term replication of Epstein-Barr virus-derived episomal vectors in the rodent cells. AB - Plasmids containing the origin of replication, oriP, of the Epstein-Barr virus (EBV) and EBV nuclear antigen-1 genes replicate extrachromosomally in primate cells. However, these plasmids have been believed not to replicate in rodent cells. We demonstrate here that these plasmids can replicate in some types of rodent cells over a long period. This result should offer not only the new insight into the mechanisms of species-specific replication of EBV, but also the possibility that an EBV-based vector can be used for gene transfer experiments in non-primate cells and an animal experiment regarding human gene therapy. PMID- 10788609 TI - Solitary HERV-K LTRs possess bi-directional promoter activity and contain a negative regulatory element in the U5 region. AB - Reporter gene analysis of HERV-K solitary long terminal repeats (LTRs) showed that they retain detectable activity in human teratocarcinoma cells, and can direct the transcription in both orientations relative to the reporter gene. Deletion analysis demonstrated the possible existence of alternative promoters within the LTR as well as a silencer-like element in the U5 region. Our results indicate also that all-trans-retinoic acid is capable of modulating expression of the reporter gene directed by a HERV-K LTR in NT2/D1 cells. PMID- 10788608 TI - The catalytic RNA of RNase P from Escherichia coli cleaves Drosophila 2S ribosomal RNA in vitro: a new type of naturally occurring substrate for the ribozyme. AB - We have found that the catalytic RNA of RNase P of Escherichia coli (M1 RNA) can cleave 2S ribosomal RNA (2S rRNA) of Drosophila melanogaster at specific positions in vitro. The cleavage mainly occurred at two sites between nucleotides 11 and 12, and between 16 and 17 of 2S rRNA. Kinetic analyses of the reaction revealed that a dimer caused by intermolecular interaction of 2S rRNA may be the substrate for the cleavage between 11 and 12, while a simple monomer is the substrate for the cleavage between 16 and 17. Substrate recognition by M1 RNA is also discussed. PMID- 10788610 TI - N-acetylcysteine suppresses TNF-induced NF-kappaB activation through inhibition of IkappaB kinases. AB - Here, we used a reductant, N-acetyl-L-cysteine (NAC), to investigate the redox sensitive step(s) in the signalling pathway from the tumor necrosis factor (TNF) receptor to nuclear factor kappaB (NF-kappaB). We found that NAC suppressed NF kappaB activation triggered by TNF or by overexpression of either the TNF receptor-associated death domain protein, TNF receptor-associated factor 2, NF kappaB-inducing kinase (NIK), or IkappaB kinases (IKKalpha and IKKbeta). NAC also suppressed the TNF-induced activation of IKKalpha and IKKbeta, phosphorylation and degradation of IkappaB, and nuclear translocation of NF-kappaB. Furthermore, NAC suppressed the activation of IKKalpha and IKKbeta triggered by the overexpression of NIK. These results indicate that IKKalpha and IKKbeta are subject to redox regulation in the cells, and that NAC inhibits NF-kappaB activation through the suppression of these kinases. PMID- 10788611 TI - Expression of a constitutively active form of phosphatidylinositol 3-kinase inhibits the induction of nitric oxide synthase in human astrocytes. AB - The present study underlines the importance of phosphatidylinositol 3-kinase (PI 3-kinase) in attenuating the induction of nitric oxide synthase (iNOS) in human astrocytes. Proinflammatory cytokines induced the production of nitric oxide (NO) and the expression of iNOS in human U373MG astrocytoma cells and primary astrocytes. Expression of a catalytically active p110 subunit (p110*) of PI 3 kinase but not that of a kinase-deficient mutant of p110 (p110-kd) induced an increase in PI 3-kinase activity and inhibited cytokine-induced production of NO and expression of iNOS. However, expression of p110* had no effect on the activation of NF-kB, suggesting that p110* inhibits the expression of iNOS without inhibiting the activation of NF-kB. PMID- 10788612 TI - Cation- and peptide-binding properties of human centrin 2. AB - Centrin and calmodulin (CaM) are closely related four-EF-hand Ca(2+)-binding proteins. While CaM is monomeric, centrin 2 is dimeric and binds only two Ca(2+) per dimer, likely to site IV in each monomer. Ca(2+) binding to centrin 2 displays pronounced negative cooperativity and a [Ca(2+)](0.5) of 30 microM. As in CaM, Ca(2+) binding leads to the exposure of a hydrophobic probe-accessible patch on the surface of centrin 2. Provided Ca(2+) is present, centrin 2 forms a 1:1 peptide:monomer complex with melittin with an affinity of 100 nM. The complex binds four instead of two Ca(2+). Our data point to surprising differences in the mode of activation of these homologous proteins. PMID- 10788613 TI - Translation termination factor aRF1 from the archaeon Methanococcus jannaschii is active with eukaryotic ribosomes. AB - Class-1 translation termination factors (release factors (RFs)) from Eukarya (eRF1) and Archaea (aRF1) exhibit a high degree of amino acid sequence homology and share many common motifs. In contrast to eRF1, function(s) of aRF1 have not yet been studied in vitro. Here, we describe for the first time the cloning and expression in Escherichia coli of the gene encoding the peptide chain RF from the hyperthermophilic archaeon Methanococcus jannaschii (MjaRF1). In an in vitro assay with mammalian ribosomes, MjaRF1, which was overproduced in E. coli, was active as a RF with all three termination codon-containing tetraplets, demonstrating the functional resemblance of aRF1 and eRF1. This observation confirms the earlier prediction that eRF1 and aRF1 form a common structural functional eRF1/aRF1 protein family, originating from a common ancient ancestor. PMID- 10788614 TI - Bacteriophage and host mutants causing the rolling-circle lambda DNA replication early after infection. AB - There are two modes of bacteriophage lambda DNA replication during its lytic development in Escherichia coli cells. The circle-to-circle (theta) replication predominates at early stages of the phage growth, whereas rolling-circle (sigma) replication occurs late after infection to produce long concatemers that serve as substrates for packaging of lambda DNA into phage proheads. The mechanism regulating the switch from theta to sigma replication remains unknown. Our previous genetic studies indicated that the bacteriophage lambda Pts1piA66 mutant cannot replicate at 43 degrees C in the wild-type E. coli host, but it can replicate in the dnaA46(ts) mutant. Density shift experiments suggested that the parental DNA molecules of the infecting phage enter sigma replication. Here, using electron microscopy, we demonstrate that as soon as 5 min after infection of the dnaA46(ts) mutant by the lambdaPts1piA66 phage at 43 degrees C, the sigma replication intermediates are highly predominant over theta replication intermediates, contrary to the wild-type conditions (wild-type bacteria infected with the lambdaP(+) phage). The initiation of replication of the lambdaPts1piA66 mutant at 43 degrees C was strongly inhibited in the dnaA(+) host, as demonstrated by electron microscopy and by pulse-labeling of the phage-derived plasmid replicon. Implications for the mechanism of the regulation of the switch from theta to sigma replication mode are discussed. PMID- 10788616 TI - Thiol modification of diacylglycerol kinase: dependence upon site membrane disposition and reagent hydrophobicity. AB - Reaction rates were determined between disulfide reagents of varying hydrophobicity and single-cysteine mutants of diacylglycerol kinase, an integral membrane protein. Polar reagents reacted most rapidly with surface-exposed sites. However, a very non-polar reagent also reacted more rapidly with exposed cysteines than with membrane sites. Moreover, this non-polar reagent usually reacted more slowly with membrane sites than did more polar reagents. These results are consistent with the notion that disulfide exchange reactions involving buried cysteines of diacylglycerol kinase are very slow in the membrane interior, such that the competing rates of reactions which occur when normally buried cysteine sites make motional excursions to hydrated regions of the interface can be significant. PMID- 10788615 TI - Modulation of the oxygen affinity of cobalt-porphyrin by globin. AB - We have combined two extreme effects which influence the oxygen affinity to obtain a cobalt-based oxygen carrier with an affinity similar to that of human adult hemoglobin (HbA). The goal was to obtain an oxygen transporter with a lower oxidation rate. Exchange of the heme group (Fe-protoporphyrin IX) in Hb with a cobalt-porphyrin leads to a reduction in oxygen affinity by over a factor of 10, an oxygen affinity too low for use as a blood substitute. At the other extreme, certain globin sequences are known to provide a very high oxygen affinity; for example, Hb Ascaris displays an oxygen affinity 1000 times higher than HbA. We demonstrate here that these opposing effects can be additive, yielding an oxygen affinity similar to that of HbA, but with oxygen binding to a cobalt atom. We have tested the effect of substitution of cobalt-porphyrin for heme in normal HbA, sperm whale (SW) Mb (Mb), and high affinity globins for leghemoglobin, two trematode Hbs: Paramphistomum epiclitum (Pe) and Gastrothylax crumenifer (Gc). As for HbA or SW Mb, the transition from heme to cobalt-porphyrin in the trematode Hbs leads to a large decrease in the oxygen affinity, with oxygen partial pressures for half saturation (P(50)) of 5 and 25 mm Hg at 37 degrees C for cobalt-Pe and cobalt-Gc, respectively. A critical parameter for Hb-based blood substitutes is the autoxidation rate; while both metals oxidize to an inactive state, we observed a decrease in the oxidation rate of over an order of magnitude for cobalt versus iron, for similar oxygen affinities. The time constants for autoxidation at 37 degrees C were 250 and 100 h for Pe and Gc, respectively. PMID- 10788617 TI - Mit1/Lb9 and Copg2, new members of mouse imprinted genes closely linked to Peg1/Mest(1). AB - Two mouse genes, Mit1/Lb9 and Copg2, linked to Peg1/Mest on mouse chromosome 6, were identified to be imprinted maternally and paternally, respectively. Mit1/Lb9 encoding untranslated transcripts resides within the intron 20 of Copg2. The gene is maternally imprinted in adult mouse brain, partially imprinted in other tissues. Copg240 kb genomic region, being expressed ubiquitously in mouse tissues with a partial imprinting pattern in embryos, neonates, and adult brain in contrast to maternally imprinted human COPG2. In addition, we identified an antisense transcript of Copg2, Copg2AS, which overlaps 3'-UTRs of Copg2 and Peg1/Mest. The Copg2AS transcript is maternally imprinted in embryos, neonates, and adult tissues. PMID- 10788619 TI - Cytotoxic activity of a recombinant GnRH-PAP fusion toxin on human tumor cell lines. AB - Pokeweed antiviral protein (PAP), a ribosome-inactivating protein isolated from the leaves of Phytolacca americana, reveals potent antiviral activity against viruses or cytotoxic action against cells once inside the cytoplasm. Therefore PAP is a good candidate to be used as an immunotoxin. We constructed a bacterial expression plasmid encoding PAP as a fusion protein with gonadotropin-releasing hormone (GnRH), a neuropeptide with receptor sites on several gynaecologic tumors. The resulting recombinant toxin was produced in Escherichia coli and accumulated in inclusion bodies. After purification under denaturing conditions, renaturated GnRH-PAP shows an IC(50) of 3 nM on in vitro translation assays and selectively inhibits the growth of the GnRH receptor positive Ishikawa cell line (ID(50) of 15 nM); on the other hand, neither GnRH nor PAP alone had any effect. PMID- 10788618 TI - SOCS-1 can suppress CD3zeta- and Syk-mediated NF-AT activation in a non-lymphoid cell line. AB - To elucidate T cell antigen receptor (TCR) signaling leading to activation nuclear factor of activated T cells (NF-AT), we reconstituted TCR signaling to activate NF-AT in a non-lymphoid cell line, 293T. We demonstrated that co expression of CD8/zeta and Syk were necessary for NF-AT activation in 293T. This NF-AT response was completely inhibited by the addition of cyclosporin A or FK506, but markedly enhanced by the additional expression of Tec protein tyrosine kinase. We also show that the cytokine signaling suppressor, suppressor of cytokine signaling 1, potently inhibited this response by interacting with Syk and immunoreceptor tyrosine-based activation motifs in CD8/zeta. These results imply that this novel system may provide a useful tool to delineate or identify the regulatory molecules for CD3zeta/Syk-mediated NF-AT activation. PMID- 10788620 TI - The C-A mismatch base pair and the single-strand terminus in the E. coli initiator tRNA(fMet) acceptor stem adopt unusual conformations. AB - Acceptor stem variants of tRNA(fMet) (Escherichia coli) have been characterized by nuclear magnetic resonance. The wild type contains a C1-A72 mismatch pair which is crucial for its biological function. For comparison, the mismatch was replaced by regular pairs U1-A72 and C1-G72. Further variants contain an altered discriminator base, G73, or a G1-C72/U73 combination. The stems of variants U1 A72/A73 and C1-G72/A73 have A-RNA geometry, which extends essentially to the single-strand terminus. C1-A72/G73 variant and wild type are structurally almost identical. C1 and A72 adopt peculiar conformations with C1 being largely destacked with respect to G2, while A73 stacks upon C1. The unique arrangement of the mismatch causes a distinctly different orientation of the single-strand terminus compared to variants with regular 1-72 base pairs, and to formyltransferase-complexed tRNA(fMet). PMID- 10788621 TI - Rad24 is essential for proliferation of diploid cells in fission yeast. AB - The rad24(+) gene of Schizosaccharomyces pombe encodes a ubiquitously expressed 14-3-3 protein. We report here that Deltarad24 cells displayed a defect in diploid colony formation, although they conjugated efficiently. We found that a cumulative deletion of mei2(+) gene almost completely suppressed this defect, and demonstrated using two-hybrid analysis that Rad24 protein directly associates with Mei2 protein by recognizing Ser-438 which is a phosphorylation target of Pat1 kinase. We conclude that constitutive progression to meiosis, caused by lack of Mei2 inhibition due to the absence of Rad24 protein, is the primary cause of the proliferative deficiency observed in Deltarad24 cells. PMID- 10788622 TI - Serum amyloid A-derived peptides, present in human rheumatic synovial fluids, induce the secretion of interferon-gamma by human CD(4)(+) T-lymphocytes. AB - Serum amyloid A (SAA) is a major acute-phase protein whose biochemical functions remain largely obscure. Human rheumatic synovial fluids were screened by high performance liquid chromatography mass spectrometry for SAA-derived peptides, specifically the sequence AGLPEKY (SAA(98-104)) which was previously shown to modulate various leukocyte functions. Two such fluids were found to contain a truncated version of SAA(98-104). Synthetic SAA(98-104) and several of its analogs were shown capable of binding isolated human CD(4)(+) T-lymphocytes and stimulating them to produce interferon-gamma. Given the high acute-phase serum level of SAA and its massive proteolysis by inflammatory related enzymes, SAA derived peptides may be involved in host defense mechanisms. PMID- 10788623 TI - Resonance Raman spectroscopy of sensory rhodopsin II from Natronobacterium pharaonis. AB - Sensory rhodopsin II (pSRII), the photophobic receptor from Natronobacterium pharaonis, has been studied by time-resolved resonance Raman (RR) spectroscopy using the rotating cell technique. Upon excitation with low laser power, the RR spectra largely reflect the parent state pSRII(500) whereas an increase of the laser power leads to a substantial accumulation of long-lived intermediates contributing to the RR spectra. All RR spectra could consistently be analysed in terms of four component spectra which were assigned to the parent state pSRII(500) and the long-lived intermediates M(400), N(485) and O(535) based on the correlation between the C = C stretching frequency and the absorption maximum. The parent state and the intermediates N(485) and O(535) exhibit a protonated Schiff base. The C = N stretching frequencies and the H/D isotopic shifts indicate strong hydrogen bonding interactions of the Schiff base in pSRII(500) and O(535) whereas these interactions are most likely very weak in N(485). PMID- 10788624 TI - Silefrin, a sodefrin-like pheromone in the abdominal gland of the sword-tailed newt, Cynops ensicauda. AB - Sodefrin-like female-attracting pheromone was purified from the abdominal glands of male sword-tailed newts, Cynops ensicauda, by gel-filtration chromatography and reversed-phase high-performance liquid chromatography. The final product comprises 10 amino acid residues with the sequence SILSKDAQLK which coincided with the sequence deduced from its precursor cDNA. This peptide was designated silefrin. The sequence of silefrin was different from that of sodefrin by two amino acid residues, with substitutions Leu for Pro and Gln for Leu at positions 3 and 8, respectively. Both native and synthetic silefrin exerted an equipotent activity in attracting conspecific females. PMID- 10788625 TI - Heat induced expression of CD95 and its correlation with the activation of apoptosis upon heat shock in rat histiocytic tumor cells. AB - The heat shock response is a universal phenomenon and is among the most highly conserved cellular responses. However, BC-8, a rat histiocytoma, fails to mount a heat shock response unlike all other eukaryotic cells. In the absence of induction of heat shock proteins, apoptotic cell death is activated in BC-8 tumor cells upon heat shock. We demonstrate here that stable transformants of BC-8 tumor cells transfected with hsp70 cDNA constitutively express hsp70 protein and are transiently protected from heat induced apoptosis for 6-8 h. In addition heat stress induces CD95 gene expression in these tumor cells. There is a delay in CD95 expression in hsp70 transfected cells suggesting a correlation between the cell surface expression of CD95 and the time of induction of apoptosis in this tumor cell line. Also expression of CD95 antigen appears to inhibit the interaction between heat shock factors and heat shock elements in these cells resulting in the lack of heat shock response. PMID- 10788626 TI - The species specificity of growth hormone requires the cooperative interaction of two motifs. AB - Primate growth hormones (GH) activate both primate and non-primate somatotrophic receptors (GH receptors), but non-primate GHs do not activate primate GH receptors. Previous studies argued the interaction of Asp(171) of human GH and Arg(43) of the receptor produced an attractive ionic interaction. In non-primate GHs, His(170) replaces the homologous Asp(171), producing a repulsive interaction with Arg(43) of the primate receptor which was believed to reduce the attraction of non-primate GH for the human GH receptor, thus providing species specificity. In this report, H170D bovine GH had activity and affinity for human GH receptors approaching those of human GH. In contrast, replacing Asp(171) of human GH with His did not significantly reduce somatotrophic activity, indicating that species specificity is not wholly explained by this residue's interaction with Arg(43) of the receptor. Deletion of either Phe(44) (a residue present only in primate GHs) or residues 32-46 (20-kDa form of human GH) each only marginally reduced somatotrophic activities. But the combination of the D171H mutation with either DeltaPhe(44) or Delta32-46 in human GH reduced binding and activity in a greater than additive fashion, indicated a functional interaction between these distant structural features. In bovine GH addition of phenylalanine at position 44 increased the somatotrophic activity and receptor affinity in cells containing the human GH receptor. The combination of the H170D mutation and the addition of phenylalanine at position 44 created a bovine GH with activity indistinguishable from wild-type human GH. Based on evidence from both bovine and human GHs, the cooperative interaction of these two distant motifs determined the species specificity and indicated that structural plasticity was a critical feature necessary for the species specificity of somatotrophic activity. PMID- 10788627 TI - The essential role of Glu-185 and Tyr-354 residues in the ferroxidase activity of Saccharomyces cerevisiae Fet3. AB - The structural determinants required for ferroxidase activity by the yeast multicopper oxidase Fet3 have been partially clarified by site-directed mutagenesis based on homology modeling. Glu-185 and Tyr-354 were substituted with Ala and Phe, respectively. Fet3 E185A retained ca. 5% residual ferroxidase catalytic efficiency, and almost 40% oxidase efficiency. On the other hand, Fet3 Y354F exhibited 50% residual efficiency as a ferroxidase and more than 70% as an oxidase. These results provide new insights in the mechanism of iron binding and oxidation by Fet3, establishing the essential role of Glu-185 and Tyr-354, and allowing to dissect ferroxidase from non-iron oxidase activity. PMID- 10788628 TI - IgA1 protease from Neisseria gonorrhoeae inhibits TNFalpha-mediated apoptosis of human monocytic cells. AB - The modulation of programmed cell death is a common theme in the patho-physiology of inflammation and infectious disease. The synthesis and secretion of an IgA1 protease is strictly associated with virulence of the Neisseria species. Here, we report on the inhibition of tumor necrosis factor alpha (TNFalpha)-mediated apoptosis of the human myelo-monocytic cell line U937 by highly purified IgA1 protease. Apoptosis was verified by the cell surface exposure of phosphatidyl serine and by terminal transferase mediated end-labeling of fragmented DNA. Interestingly, IgA1 protease specifically cleaved the TNF receptor II (TNF-RII) on the surface of intact cells whereas TNF-RI was not affected by the enzyme. Therefore, inhibition of TNFalpha-mediated apoptosis might be correlated to specific cleavage of the TNF-RII by neisserial IgA1 protease. PMID- 10788629 TI - Characterization of tBid-induced cytochrome c release from mitochondria and liposomes. AB - tBid, the cleaved form of Bid, can induce cytochrome c (Cyt. c) release from rat heart mitochondria more efficiently and reproducibly than that from liver or brain mitochondria. Unlike Bax, such release was not prevented by cyclosphorin A, an inhibitor of the opening of permeability transition pore. Carbonyl-cyanide m chlorophenyl-hydrazone or oligomycin also have no obvious effect on the release of Cyt. c. In contrast to ceramide, tBid-mediated Cyt. c release from mitochondria is independent of the redox state of Cyt. c. Furthermore, Bid or tBid can directly trigger the efflux of encapsulated Cyt. c or trypsin within liposomes without involvement of other protein factors. PMID- 10788630 TI - Thrombin-induced inhibition of myoblast differentiation is mediated by Gbetagamma. AB - Thrombin has been shown to inhibit skeletal muscle differentiation. However, the mechanisms by which thrombin represses myogenesis remain unknown. Since the thrombin receptor couples to G(i), G(q/11) and G(12), we examined which subunits of heterotrimeric guanine nucleotide-binding regulatory proteins (Galpha(i), Galpha(q/11), Galpha(12) or Gbetagamma) participate in the thrombin-induced inhibition of C2C12 myoblast differentiation. Galpha(i2) and Galpha(11) had no inhibitory effect on the myogenic differentiation. Galpha(12) prevented only myoblast fusion, whereas Gbetagamma inhibited both the induction of skeletal muscle-specific markers and the myotube formation. In addition, the thrombin induced reduction of creatine kinase activity was blocked by the C-terminal peptide of beta-adrenergic receptor kinase, which is known to sequester free Gbetagamma. These results suggest that the thrombin-induced inhibition of muscle differentiation is mainly mediated by Gbetagamma. PMID- 10788631 TI - The compositional organization and the expression of the Arabidopsis genome. AB - The base composition patterns of genes, coding sequences and gene expression levels were analyzed in the available long sequences (contigs) of Arabidopsis. Chromosome 5 was analyzed in detail and all chromosomes for which sequence data are now available show essentially the same large-scale compositional properties. Guanine+cytosine levels of genes and of their coding regions, as well as gene densities and expression levels, all show a marked tendency to be higher in the distal regions of Arabidopsis chromosomes. PMID- 10788632 TI - Mutagenic analysis of Thr-232 in rhodanese from Azotobacter vinelandii highlighted the differences of this prokaryotic enzyme from the known sulfurtransferases. AB - Azotobacter vinelandii RhdA uses thiosulfate as the only sulfur donor in vitro, and this apparent selectivity seems to be a unique property among the characterized sulfurtransferases. To investigate the basis of substrate recognition in RhdA, we replaced Thr-232 with either Ala or Lys. Thr-232 was the target of this study since the corresponding Lys-249 in bovine rhodanese has been identified as necessary for catalytic sulfur transfer, and replacement of Lys-249 with Ala fully inactivates bovine rhodanese. Both T232K and T232A mutants of RhdA showed significant increase in thiosulfate-cyanide sulfurtransferase activity, and no detectable activity in the presence of 3-mercaptopyruvate as the sulfur donor substrate. Fluorescence measurements showed that wild-type and mutant RhdAs were overexpressed in the persulfurated form, thus conferring to this enzyme the potential of a persulfide sulfur donor compound. RhdA contains a unique sequence stretch around the catalytic cysteine, and the data here presented suggest a possible divergent physiological function of A. vinelandii sulfurtransferase. PMID- 10788633 TI - Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus. AB - Transient kinetics of the equine infectious anemia virus deoxyuridine 5' triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis. PMID- 10788634 TI - Pre-attentive and attentive mechanisms in vision. Perceptual organization and dysfunction. PMID- 10788635 TI - Salience from feature contrast: additivity across dimensions. AB - Test targets ('singletons') that displayed orientation, motion, luminance, or color contrast, or pairwise combinations of these, were presented in line texture arrays, and their saliences were quantified in comparison to reference targets at defined luminance levels. In general, saliency effects in different stimulus dimensions did add, but did not add linearly. That is, targets with feature contrast in two dimensions were generally more salient than targets with only one of these properties, but often less salient than predicted from the sum of the individual saliency components. Salience variations within a dimension were compared with and without a second saliency effect added. The resulting gain reduction in the combined stimulus conditions was interpreted to reflect the amount of overlap between the respective saliency mechanisms. Combinations of orientation and color contrast produced the strongest gain reduction (about 90% for color in orientation) thus indicating the strongest overlap of underlying saliency mechanisms. Combinations of orientation and motion contrast revealed about 50% overlap, slightly smaller rates were found for combinations of color and motion. All combinations with luminance contrast (orientation and luminance, motion and luminance) produced only little gain reduction (<30%) thus indicating a higher degree of independence between the underlying saliency mechanisms than for other stimulus dimensions. PMID- 10788637 TI - Interactions between attention, context and learning in primary visual cortex. AB - Attention in early visual processing engages the higher order, context dependent properties of neurons. Even at the earliest stages of visual cortical processing neurons play a role in intermediate level vision - contour integration and surface segmentation. The contextual influences mediating this process may be derived from long range connections within primary visual cortex (V1). These influences are subject to perceptual learning, and are strongly modulated by visuospatial attention, which is itself a learning dependent process. The attentional influences may involve interactions between feedback and horizontal connections in V1. V1 is therefore a dynamic and active processor, subject to top down influences. PMID- 10788636 TI - Spatial covert attention increases contrast sensitivity across the CSF: support for signal enhancement. AB - This study is the first to report the benefits of spatial covert attention on contrast sensitivity in a wide range of spatial frequencies when a target alone was presented in the absence of a local post-mask. We used a peripheral precue (a small circle indicating the target location) to explore the effects of covert spatial attention on contrast sensitivity as assessed by orientation discrimination (Experiments 1-4), detection (Experiments 2 and 3) and localization (Experiment 3) tasks. In all four experiments the target (a Gabor patch ranging in spatial frequency from 0.5 to 10 cpd) was presented alone in one of eight possible locations equidistant from fixation. Contrast sensitivity was consistently higher for peripherally- than for neutrally-cued trials, even though we eliminated variables (distracters, global masks, local masks, and location uncertainty) that are known to contribute to an external noise reduction explanation of attention. When observers were presented with vertical and horizontal Gabor patches an external noise reduction signal detection model accounted for the cueing benefit in a discrimination task (Experiment 1). However, such a model could not account for this benefit when location uncertainty was reduced, either by: (a) Increasing overall performance level (Experiment 2); (b) increasing stimulus contrast to enable fine discriminations of slightly tilted suprathreshold stimuli (Experiment 3); and (c) presenting a local post-mask (Experiment 4). Given that attentional benefits occurred under conditions that exclude all variables predicted by the external noise reduction model, these results support the signal enhancement model of attention. PMID- 10788638 TI - The psychophysics of visual search. AB - Most theories of visual search emphasize issues of limited versus unlimited capacity and serial versus parallel processing. In the present article, we suggest a broader framework based on two principles, one empirical and one theoretical. The empirical principle is to focus on conditions at the intersection of visual search and the simple detection and discrimination paradigms of spatial vision. Such simple search conditions avoid artifacts and phenomena specific to more complex stimuli and tasks. The theoretical principle is to focus on the distinction between high and low threshold theory. While high threshold theory is largely discredited for simple detection and discrimination, it persists in the search literature. Furthermore, a low threshold theory such as signal detection theory can account for some of the phenomena attributed to limited capacity or serial processing. In the body of this article, we compare the predictions of high threshold theory and three versions of signal detection theory to the observed effects of manipulating set size, discriminability, number of targets, response bias, external noise, and distractor heterogeneity. For almost all cases, the results are inconsistent with high threshold theory and are consistent with all three versions of signal detection theory. In the Discussion, these simple theories are generalized to a larger domain that includes search asymmetry, multidimensional judgements including conjunction search, response time, search with multiple eye fixations and more general stimulus conditions. We conclude that low threshold theories can account for simple visual search without invoking mechanisms such as limited capacity or serial processing. PMID- 10788639 TI - Mechanisms of perceptual attention in precuing of location. AB - What are the mechanisms of spatial attention underlying precue validity effects? We answer this question within the framework of a perceptual template model (PTM) [Lu & Dosher (1998). External noise distinguishes attention mechanisms. Vision Research, 38, 1183-1198; Dosher & Lu (1999). Mechanisms of perceptual learning. Vision Research, 39, 3197-3221] and an external noise plus attention paradigm for orientation judgments in two- to eight-location displays. Attentional mechanisms correspond to behavioral signatures: External noise exclusion produces cuing effects in high external noise and stimulus enhancement produces cuing effects in noiseless displays. We found that external noise exclusion was the primary mechanism of cue validity effects, with large effects in high-noise displays. Stimulus enhancement coexisted as a secondary mechanism in noiseless displays for a subset of observers and display conditions. Contrast threshold ratio tests ruled out attentionally mediated changes in gain control. The ratio rules were also shown to hold for a stochastic PTM model. Effects were equivalent for four alternative (Experiment 1) and two-alternative (Experiment 2) orientation identification. Precues allow observers to reduce noise and focus on the target in the precued location. External noise exclusion was more important in larger displays. Previous results are reclassified and understood within the PTM framework. PMID- 10788640 TI - Feature-based integration of orientation signals in visual search. AB - We have measured orientation discrimination in the presence of a variable number of neutral distracters for two distinct tasks: identification of the orientation of a tilted target and location of its position. Both tasks were performed in the presence of visual noise of variable contrasts. Under a range of conditions, subjects could identify the direction of target tilt at thresholds well below those necessary to locate its position. The location thresholds showed only weak dependency on set-size, consistent with a stimulus uncertainty of parallel search of the output of independent orientation analysers, while the identification thresholds showed a much stronger dependency, varying with the square root of set size over a wide range noise contrasts. The square root relationship suggests perceptual summation of target and distracters. Manipulating the spread of visual noise suggests that the summation is feature-based, possibly operating on the outputs of first-stage orientation analysers. Pre-cueing the target eliminates the effects of set-size, showing that the summation is under rapid attentional control; the visual system can choose between high performance over a limited area and poorer performance over a much larger area. PMID- 10788641 TI - Human development of perceptual organization. AB - Two relevant dimensions are revealed within which developmental patterns of perceptual organization might be investigated. Within the local-integrative dimension, employing a contour integration task, we found indications that spatial integration develops slowly. We also found reduced contextual modulation of a local target in children employing the Ebbinghaus illusion. Within the action-perception dimension, we hypothesize a relatively slow development of the perceptual system (mediated by the ventral visual stream), as compared to the development of the action system (mediated by the dorsal visual stream). Taken together, the data indicate that long-range neuronal connectivity supporting perceptual organization in the posterior pole of the brain, and in the ventral visual pathway is not fully developed in young children. PMID- 10788643 TI - Reappraising the apparent costs of attending to two separate visual objects. AB - Support for object-based accounts of visual attention has been drawn from several different types of effect. One effect is found when observers try to restrict their attention to a particular region of a display. Other regions belonging to the same object are often selected as well, suggesting that attention spreads spatially over entire objects. Another effect is found when judging two visual attributes; performance is often less efficient when the attributes belong to separate objects rather than both belonging to a single object. This latter effect has been taken to imply that only one segmented object can be attended at a time. However, it may instead merely be a variant of the first effect. If, as we assume here, attention spreads to task-irrelevant regions of relevant objects, it will encompass a larger spatial region and more information when judging attributes of two objects rather than one. Here we compared judging one versus two objects, while manipulating whether the two objects occupied a wider extent than the single object condition (as in previous work), or not. Costs were found for judging two objects versus one only when together they occupied a wider spatial extent. We conclude that reported difficulties in attending two objects may be due to attention spreading across the entire spatial extent of objects when judging their parts, rather than a fixed inability to process more than object at a time. PMID- 10788642 TI - Cortical image density determines the probability of target discovery during active search. AB - An analysis of target detection as a function of target eccentricity was made on eye movement data collected from three monkey subjects during active visual search. Target detection probability was invariant across array set size and eccentricity conditions when the cortical density of relevant stimuli surrounding the target was held constant. When target color was used to guide search, the effective cortical density was the density of stimuli that shared the target's color. Thus the passive constraint of cortical magnification in combination with an active selection for a stimulus attribute, in this case color, sets the spatial framework for detection of the target. PMID- 10788644 TI - The perceptual organization of visual objects: a microgenetic analysis. AB - Primed matching was used to examine the microgenesis of perceptual organization for line configurations that vary in the connectedness between their four line components, and for hierarchical patterns composed of four outline closed figures. The results for the line configurations showed that the configural organization of the disconnected line segments was available for priming very early, and its effect outweighed possible effects of the line components. An early relative dominance of the components was observed for the stimuli whose components were closed figures. These results suggest that uniform connectedness is not necessary for the designation of entry-level units. Disconnected line segments are rapidly organized into configurations, provided the presence of collinearity and/or closure. Closed figural elements are individuated early and are grouped into higher-level units with time. PMID- 10788645 TI - The spread of attention and learning in feature search: effects of target distribution and task difficulty. AB - We examined the roles of two determinants of spatial attention in governing the spread of perceptual learning, namely, stimulus location distribution and task difficulty. Subjects were trained on detection of a target element with an odd orientation imbedded in an array of light bars with otherwise uniform orientation. To assess the effects of target distribution on attention and learning, target positions were distributed so that attention was allocated not only to the target positions themselves, but also to intermediate positions where the target was not presented. Target detection performance substantially improved and improvement spread to match the induced window of spatial attention rather than only the actual target locations. To assess the effect of task difficulty on the spread of attention and learning, the target-distractor orientation difference and the time interval available for processing were manipulated. In addition, we compared performance of subjects with more versus with less detection difficulty. A consistent pattern emerged: When the task becomes more difficult, the window of attention shrinks, and learning becomes more localized. We conclude that task-specific spatial attention is both necessary and sufficient to induce learning. The spread of spatial attention, and thus of learning, is determined by the integrated effects of target distribution and task difficulty. We propose a theoretical framework whereby these factors combine to determine the cortical level of the focus of attention, which in turn enables learning modifications. PMID- 10788646 TI - Tracking the apparent location of targets in interpolated motion. AB - Under appropriate conditions, a target moving in discrete steps can appear to move smoothly and continuously even within the portions of the path where no physical stimulus is present. We investigated the nature of this interpolated motion in attentive tracking displays as well as apparent motion. The results showed that the apparent location of the target moved smoothly through space between the two discrete locations and the judgements of interpolated motion for attentive tracking and apparent motion were comparable to those for continuous motion in both the perceived path and the precision of the judgements. There were few, if any, differences between judgements for real and interpolated motion. An alignment procedure showed that the smooth change in location judgements was real and not a consequence of averaging across discrete locations actually seen on each trial. We also found that the slowest alternation rate which supported accurate location judgements corresponded to a critical SOA of about 500 ms, similar to the longest SOA which supported a subjective impression of motion in the display. Deviations from a constant velocity which were shorter than 200 ms did not register in the judged motion path, suggesting a fairly long time constant for the integration of velocity information into the perceived motion. These results suggest a specialized motion analysis which provides an accurate, explicit model of the interpolated motion path. PMID- 10788647 TI - Failure to detect changes in color for lines rotating in depth: the effects of grouping and type of color change. AB - A new technique for measuring change detection was introduced in which contours rotating in depth around a vertical axis (in a computer display) could be altered in color as they passed through their point of minimum extension (the median plane) where a thin static vertical occluder hid the change. Sets of five or six contours were either strongly grouped (similar in length, orientation and spacing) or weakly grouped (of variable length, orientation and spacing). Changes consisted of one line changing to a new color or else two lines swapping colors. The measure was the proportion of missed changes. When subjects were not instructed to look for change almost no changes were reported although subjects were told beforehand that they would have to describe the configuration after viewing it. When subjects were instructed to look for changes, it was found that detection of color change was significantly better for strongly grouped lines. It is proposed that grouping, by reducing redundancy, also reduces attentional demands with respect to the properties on which it is based, making it easier to attend to and therefore detect changes in other properties. We found that it was much easier to detect the introduction of a new color than to detect a swap between two existing colors. It is hypothesized that swap-type changes were harder to detect because they required attention to a conjunction of position and color. PMID- 10788648 TI - The implementation of visual routines. AB - Many visual tasks can be decomposed into a sequence of simpler subtasks. Ullman suggested that such subtasks are carried out by elemental operations that are implemented by specialized processes in the visual brain [Ullman, S. (1984). Visual routines. Cognition (18), 97-159]. According to this hypothesis, there are a limited number of elemental operations that, since they can be applied sequentially, may nevertheless give rise to a large number of visual routines. Examples of such elemental operations are visual search, texture segregation and contour grouping. Here we attempt to delineate how such elemental operations are implemented in the visual brain. When an image appears, feedforward processing rapidly leads to an activity pattern that is distributed across many visual areas. Thereafter, elemental operations come into play, and these are implemented by the modulation of firing rates. Firing rate modulations effectuate grouping of neural responses into coherent object representations. Moreover, they permit transfer of information from one operator to the next, which allows flexibility in the sequencing of operations. We discuss how the elemental operations provide a tool to relate cortical physiology to psychophysics, and suggest a reclassification of pre-attentive and attentive processes. PMID- 10788649 TI - Contrast-sensitive perceptual grouping and object-based attention in the laminar circuits of primary visual cortex. AB - Recent neurophysiological studies have shown that primary visual cortex, or V1, does more than passively process image features using the feedforward filters suggested by Hubel and Wiesel. It also uses horizontal interactions to group features preattentively into object representations, and feedback interactions to selectively attend to these groupings. All neocortical areas, including V1, are organized into layered circuits. We present a neural model showing how the layered circuits in areas V1 and V2 enable feedforward, horizontal, and feedback interactions to complete perceptual groupings over positions that do not receive contrastive visual inputs, even while attention can only modulate or prime positions that do not receive such inputs. Recent neurophysiological data about how grouping and attention occur and interact in V1 are simulated and explained, and testable predictions are made. These simulations show how attention can selectively propagate along an object grouping and protect it from competitive masking, and how contextual stimuli can enhance or suppress groupings in a contrast-sensitive manner. PMID- 10788650 TI - Intention-related activity in the posterior parietal cortex: a review. AB - Over the last few years it is becoming increasingly apparent that an important role of the posterior parietal cortex is to process sensory information for the purpose of planning actions. We review studies showing that a large component of neural activity in area LIP is related to planning saccades and activity in a nearby parietal reach region (PRR) to reaches. This intention related activity dominates the delay period in delayed movement tasks, and also comprises a substantial component of the transient response. These findings, along with additional anatomical and physiological evidence, lends support to the idea that different cortical areas within the PPC represent plans for different actions. We also found strong modulation of activity when movement plans were changed without changes in the locus of attention. This result suggests that PPC, which has been postulated to play a role in shifting attention, may also play a role in changing movement intentions. Sensory related activity was also present in these tasks and may be related to the stimulus or to attention. These experiments show that there are intention and sensory related activities in the PPC consistent with its proposed role in sensory-motor transformations. These studies also show that care must be taken to measure intention-related signals and not assume that all task dependent modulation in the PPC reflects attention. PMID- 10788651 TI - Attentional and oculomotor capture by onset, luminance and color singletons. AB - In three experiments we investigated whether attentional and oculomotor capture occur only when object-defining abrupt onsets are used as distractors in a visual search task, or whether other salient stimuli also capture attention and the eyes even when they do not constitute new objects. The results showed that abrupt onsets (new objects) are especially effective in capturing attention and the eyes, but that luminance increments that do not accompany the appearance of new objects capture attention as well. Color singletons do not capture attention unless subjects have experienced the color singleton as a search target in a previous experimental session. Both abrupt onsets and luminance increments elicit reflexive, involuntary saccades whereas transient color changes do not. Implications for theories of attentional capture are discussed. PMID- 10788652 TI - The lateral intraparietal area as a salience map: the representation of abrupt onset, stimulus motion, and task relevance. AB - Neurons in the lateral intraparietal area (LIP) of the monkey represent salient stimuli. They respond to recently flashed stimuli that enter their receptive fields by virtue of saccades better than they respond to stable, behaviorally irrelevant stimuli brought into their receptive fields by saccades. They respond transiently to abrupt motion onsets, but have no directional selectivity. They respond to stable stimuli that are the targets for saccadic eye movements, but far less before the same saccades without stimuli. LIP is important in the attentional mechanisms preceding the choice of saccade target rather than in the intention to generate the saccade itself. PMID- 10788653 TI - Seeing, sensing, and scrutinizing. AB - Large changes in a scene often become difficult to notice if made during an eye movement, image flicker, movie cut, or other such disturbance. It is argued here that this change blindness can serve as a useful tool to explore various aspects of vision. This argument centers around the proposal that focused attention is needed for the explicit perception of change. Given this, the study of change perception can provide a useful way to determine the nature of visual attention, and to cast new light on the way that it is - and is not - involved in visual perception. To illustrate the power of this approach, this paper surveys its use in exploring three different aspects of vision. The first concerns the general nature of seeing. To explain why change blindness can be easily induced in experiments but apparently not in everyday life, it is proposed that perception involves a virtual representation, where object representations do not accumulate, but are formed as needed. An architecture containing both attentional and nonattentional streams is proposed as a way to implement this scheme. The second aspect concerns the ability of observers to detect change even when they have no visual experience of it. This sensing is found to take on at least two forms: detection without visual experience (but still with conscious awareness), and detection without any awareness at all. It is proposed that these are both due to the operation of a nonattentional visual stream. The final aspect considered is the nature of visual attention itself - the mechanisms involved when scrutinizing items. Experiments using controlled stimuli show the existence of various limits on visual search for change. It is shown that these limits provide a powerful means to map out the attentional mechanisms involved. PMID- 10788654 TI - A saliency-based search mechanism for overt and covert shifts of visual attention. AB - Most models of visual search, whether involving overt eye movements or covert shifts of attention, are based on the concept of a saliency map, that is, an explicit two-dimensional map that encodes the saliency or conspicuity of objects in the visual environment. Competition among neurons in this map gives rise to a single winning location that corresponds to the next attended target. Inhibiting this location automatically allows the system to attend to the next most salient location. We describe a detailed computer implementation of such a scheme, focusing on the problem of combining information across modalities, here orientation, intensity and color information, in a purely stimulus-driven manner. The model is applied to common psychophysical stimuli as well as to a very demanding visual search task. Its successful performance is used to address the extent to which the primate visual system carries out visual search via one or more such saliency maps and how this can be tested. PMID- 10788655 TI - The role of primary visual cortex (V1) in visual awareness. AB - In the search for the neural correlate of visual awareness, much controversy exists about the role of primary visual cortex. Here, the neurophysiological data from V1 recordings in awake monkeys are examined in light of two general classes of models of visual awareness. In the first model type, visual awareness is seen as being mediated either by a particular set of areas or pathways, or alternatively by a specific set of neurons. In these models, the role of V1 seems rather limited, as the mere activity of V1 cells seems insufficient to mediate awareness. In the second model type, awareness is hypothesized to be mediated by a global mechanism, i.e. a specific kind of activity not linked to a particular area or cell type. Two separate versions of global models are discussed, synchronous oscillations and spike rate modulations. It is shown that V1 synchrony does not reflect perception but rather the horizontal connections between neurons, indicating that V1 synchrony cannot be a direct neural correlate of conscious percepts. However, the rate of spike discharges of V1 neurons is strongly modulated by perceptual context, and these modulations correlate very well with aspects of perceptual organization, visual awareness, and attention. If these modulations serve as a neural correlate of visual awareness, then V1 contributes to that neural correlate. Whether V1 plays a role in the neural correlate of visual awareness thus strongly depends on the way visual awareness is hypothesized to be implemented in the brain. PMID- 10788656 TI - Antecedents and correlates of visual detection and awareness in macaque prefrontal cortex. AB - We have investigated the neural basis of visual detection in monkeys trained to report the presence or absence of a visual stimulus that was rendered intermittently detectable by backward masking. Neurons were recorded in the frontal eye field (FEF), an area located in prefrontal cortex that is involved in converting the outcome of visual processing into a command to shift gaze. The behavioral and neuronal data were analyzed in terms of signal detection theory. We found that the initial visual responses in FEF provided signals that could form the basis for correct or erroneous detection of the target. A later phase of prolonged elevated activity occurred in many visual neurons and all movement neurons that was highly correlated with the monkey's report of target presence. When observed in movement cells that project to oculomotor structures, this period of activation is interpreted as a motor command leading to the behavioral response. When observed in visual cells that do not project to oculomotor structures, the later period of activation does not admit to the motor command interpretation. Because the visual neurons likely contribute to the feedback pathway to visual cortical areas, we hypothesize that the later selective activation in the prefrontal visual neurons interacts with ongoing activity in visual cortical areas contributing to the process by which a particular sensory representation receives enhanced activation and thereby engages attention and awareness. PMID- 10788657 TI - The importance of sustained attention for patients with maculopathies. AB - Sustained attention enhances perception in eccentric positions in the visual field, which helps patients with foveal vision loss to develop a peripheral 'preferred retinal locus' (PRL). Besides central scotoma topography, local variations of attentional performance could influence the choice of PRL location. We tested sustained attention augmenting peripheral letter recognition in 23 maculopathy patients and 15 normally-sighted subjects (eight positions, 8 degrees eccentricity). Performance was shown to depend on tested location, which was the same in patients and normals. This indicates that the choice of the PRL location after foveal vision loss can be influenced by topographic features of sustained attention. PMID- 10788658 TI - Object memory effects on figure assignment: conscious object recognition is not necessary or sufficient. AB - In three experiments we investigated whether conscious object recognition is necessary or sufficient for effects of object memories on figure assignment. In experiment 1, we examined a brain-damaged participant, AD, whose conscious object recognition is severely impaired. AD's responses about figure assignment do reveal effects from memories of object structure, indicating that conscious object recognition is not necessary for these effects, and identifying the figure ground test employed here as a new implicit test of access to memories of object structure. In experiments 2 and 3, we tested a second brain-damaged participant, WG, for whom conscious object recognition was relatively spared. Nevertheless, effects from memories of object structure on figure assignment were not evident in WG's responses about figure assignment in experiment 2, indicating that conscious object recognition is not sufficient for effects of object memories on figure assignment. WG's performance sheds light on AD's performance, and has implications for the theoretical understanding of object memory effects on figure assignment. PMID- 10788659 TI - Fractionating the binding process: neuropsychological evidence distinguishing binding of form from binding of surface features. AB - We present neuropsychological evidence demonstrating that the binding of form elements into shapes dissociates from the binding of surface detail to shape. Data are reported from a patient with bilateral parietal lesions, GK, who manifests left-side visual extinction along with many illusory conjunctions when asked to discriminate both surface and form information about stimuli. We show that there are effects of grouping on both extinction and illusory conjunctions when the tasks require report of object shape. In contrast, illusory conjunctions involving surface and form information were unaffected by grouping based on shape. In addition, grouping was stronger when forms were presented within the same hemifield than when they appeared in different hemifields, whilst illusory conjunctions of form and colour occurred equally often within and across hemifields. These results support a two-stage account of visual binding: form elements are first bound together locally into shapes, and this is followed by a second stage of binding in which shapes are integrated with surface details. The second but not the first stage of binding is impaired in this patient. PMID- 10788660 TI - Independent effects of pictorial displays on perception and action. AB - Although pictorial illusions have been used to study perception for a long time, the effects of such displays on the visual control of actions has recently been the matter of some debate. Evidence from a re-analysis of an earlier study is presented that suggests pictorial displays can exert opposite effects on perceptual size judgements and grip scaling, perhaps because the two-dimensional elements surrounding the target for a grasp are treated as potential obstacles. This interpretation was supported by the results of an experiment in which the relative position and distance of two-dimensional elements flanking a target had differential effects on perceptual judgements of size and the scaling of grip aperture. PMID- 10788661 TI - The effects of anterior arcuate and dorsomedial frontal cortex lesions on visually guided eye movements in the rhesus monkey: 1. Single and sequential targets. AB - This study examined the effects of anterior arcuate and dorsomedial frontal cortex lesions on visually guided eye movements in three rhesus monkeys. Lesions of the anterior bank of the arcuate, where the frontal eye fields reside, produced major deficits in the execution of saccadic eye movements to sequentially presented targets that did not recover even after 1 year after the lesions. Ablations of the dorsomedial frontal cortex, wherein the medial eye fields reside, produced much smaller and shorter-duration deficits on this task. Deficits after paired lesions of the anterior arcuate and dorsomedial frontal cortex were of approximately the same magnitude as after single anterior arcuate lesions. Anterior arcuate lesions also increased saccadic reaction times to single visual targets and decreased saccadic velocities that recovered gradually over a period of 2-5 months. Dorsomedial frontal cortex lesions produced only small increases in saccadic latencies that recovered rapidly. None of the lesions produced deficits in executing combined saccadic and pursuit eye movements to moving targets. The results suggest that the anterior arcuate area plays a central role in the execution of sequences of eye movements to successively appearing targets. PMID- 10788662 TI - The effects of anterior arcuate and dorsomedial frontal cortex lesions on visually guided eye movements: 2. Paired and multiple targets. AB - This study examined the effects of anterior arcuate and dorsomedial frontal cortex lesions on the execution of saccadic eye movements made to paired and multiple targets in rhesus monkeys. Identical paired targets were presented with various temporal asynchronies to determine the temporal offset required to yield equal probability choices to either target. In the intact animal equal probability choices were typically obtained when the targets appeared simultaneously. After unilateral anterior arcuate lesions a major shift arose in the temporal offset required to obtain equal probability choices for paired targets that necessitated presenting the target in the hemifield contralateral to the lesion more than 100 ms prior to the target in the ipsilateral hemifield. This deficit was still pronounced 1 year after the lesion. Dorsomedial frontal cortex lesions produced much smaller but significant shifts in target selection that recovered more rapidly. Paired lesions produced deficits similar to those observed with anterior arcuate lesions alone. Major deficits were also observed on a multiple target temporal discrimination task after anterior arcuate but not after dorsomedial frontal cortex lesions. These results suggest that the frontal eye fields that reside in anterior bank of the arcuate sulcus play an important role in temporal processing and in target selection. Dorsomedial frontal cortex, that contains the medial eye fields, plays a much less important role in the execution of these tasks. PMID- 10788663 TI - A technique to prevent dural adhesions to chronically implanted microelectrode arrays. AB - Minimizing relative movements between neural tissues and arrays of microelectrodes chronically implanted into them is expected to greatly enhance the capacity of the microelectrodes to record from single cortical neurons on a long-term basis. We describe a new surgical technique to minimize the formation of adhesions between the dura and an implanted electrode array using a 12 microm (0.5 mil) thick sheet of Teflon film positioned between the array and the dura. A total of 15 cats were implanted using this technique. Gross examination of 12 implant sites at the time of sacrifice failed to find evidence of adhesions between the arrays and the dura when the Teflon(R) film remained in its initial position. In six implants from which recordings were made, an average of nine of the 11 (81%) connected electrodes in each array recorded evoked neural activity after 180 days post implantation. Further, on average, two separable units were identified on each of the implanted electrodes in these arrays. No significant change was found in the density of cell bodies around implanted electrodes of four of the implanted electrode arrays. However, histological evaluation of the implant sites revealed evidence of meningeal proliferation beneath the arrays. The technique described is shown to be effective at preventing adhesions between implanted electrode arrays and improve the characteristics of chronic recordings obtained with these structures. PMID- 10788665 TI - Automatic morphometry of nerve histological sections. AB - A method for the automatic segmentation, recognition and measurement of neuronal myelinated fibers in nerve histological sections is presented. In this method, the fiber parameters i.e. perimeter, area, position of the fiber and myelin sheath thickness are automatically computed. Obliquity of the sections may be taken into account. First, the image is thresholded to provide a coarse classification between myelin and non-myelin pixels. Next, the resulting binary image is further simplified using connected morphological operators. By applying semantic rules to the zonal graph axon candidates are identified. Those are either isolated or still connected. Then, separation of connected fibers is performed by evaluating myelin sheath thickness around each candidate area with an Euclidean distance transformation. Finally, properties of each detected fiber are computed and false positives are removed. The accuracy of the method is assessed by evaluating missed detection, false positive ratio and comparing the results to the manual procedure with sampling. In the evaluated nerve surface, a 0.9% of false positives was found, along with 6.36% of missed detections. The resulting histograms show strong correlation with those obtained by manual measure. The noise introduced by this method is significantly lower than the intrinsic sampling variability. This automatic method constitutes an original tool for morphometrical analysis. PMID- 10788664 TI - Assessing spatial vision - automated measurement of the contrast-sensitivity function in the hooded rat. AB - The contrast-sensitivity function (CSF) provides a concise and thorough description of an organism's spatial vision; it is widely used to describe vision in animals and humans, to track developmental changes in vision, and to compare vision among different species. Despite the predominance of rats in neuroscience research, their vision is not thoroughly studied due to the complexity of psychophysical measurement and a generally held notion that rat vision is poor. We therefore designed an economical and rapid method to assess the hooded rat's CSF, using a computer monitor to display stimuli and an infrared touch screen to record responses. A six-alternative forced-choice task presented trials in which a sine-wave grating (S+), varying in spatial frequency and contrast, was displayed at different locations along with five gray stimuli (S-). Nose pokes to the S+ but not the S- produced water reinforcers. Contrasts were tested at each spatial frequency with a simple adaptive procedure until stimulus detection fell below chance. Psychometric functions were obtained by maximum-likelihood fitting of a logistic function to the raw data, obtaining the threshold as the function's point of inflection. As in previous studies with rats, CSFs showed an inverse-U shape with peak sensitivity at 0.12 cyc/deg and acuity just under 1 cyc/deg. The results indicate the present computer-controlled behavioral testing device is a precise and efficient instrument to assess spatial visual function in rats. PMID- 10788666 TI - Fluorescein-labeled naloxone binding to mu opioid receptors on live Chinese hamster ovary cells using confocal fluorescent microscopy. AB - A general method of confocal laser scanning microscopy was used to demonstrate specific binding of fluorescein-labeled naloxone (FNAL, 10-50 nM) to stably transfected mu opioid receptors on live Chinese hamster ovary cells. Nonspecific binding was visually indistinguishable from autofluorescence in cells with intact cell membranes. Fluorescent labeling of cell perimeters, not present in control nontransfected cells, reversed in transfected cells upon washout of FNAL or following the addition of either unlabeled naloxone (25 microM) or the mu specific antagonist CTOP (1 microM). The addition of the delta and kappa specific agonists DPDPE (1 microM) and U50488 (1 microM), respectively, failed to reverse the labeling. Further evidence of specific binding was obtained from kinetic experiments, where it was observed that only transfected cells showed a time dependent exponential change in fluorescence that permitted estimation of association and dissociation binding rate constants of (5.8+/-0.5, mean+/ S.E.M.)x10(5) M(-1) s(-1) and (3.3+/-0.6)x10(-3) s(-1), respectively and a kinetically derived dissociation constant of 5.7+/-1.4 nM. These estimates were comparable to those obtained under similar conditions in radioligand binding experiments using [3H]-naloxone. PMID- 10788667 TI - Requirements for obtaining unbiased estimates of neuronal numbers in frozen sections. AB - The use of frozen sectioning is a convenient and rapid means of observing the results obtained using fluorescent retrograde tracers. Quantitation of these results using the biased stereological methods currently available can be associated with large errors. A recently developed stereological tool, the optical dissector, provides unbiased and efficient results, however, the requirements for its use in frozen sections has not previously been established. In this study, a comparison was made of neuron numbers, estimated using the optical dissector method, in the motoneuron pool retrogradely labelled from the rat sciatic nerve with either Fast Blue or Tetramethylrhodamine dextran (fluoro ruby) in methacrylate embedded and frozen spinal cord specimens. Despite over 50% shrinkage in the frozen sections, compared with virtually no shrinkage in the methacrylate sections, no significant difference in labelled motoneuron numbers was observed, provided this shrinkage was taken into account. Correction for section shrinkage is therefore essential in order to use the optical dissector with confidence to count fluorescent labelled neurons in frozen tissue. PMID- 10788668 TI - Rapid microplate assay for superoxide scavenging efficiency. AB - Here we report a method to determine superoxide scavenging efficiency, using kinetic analysis of cytochrome c reduction and an automated UV/vis microtiter plate reader. Superoxide (O(2)(-&z. rad;)) was generated by xanthine oxidase metabolism of hypoxanthine, and quantified by following reduction of cytochrome c by O(2)(-&z. rad;) as increasing absorbance at 550 nm. Reaction conditions were established that provided a linear increase in O(2)(-&z.rad;) generation for more than 20 min, and good reproducibility over time. The majority of cytochrome c reduction was blocked by superoxide dismutase, indicating cytochrome c reduction derived predominantly from O(2)(-&z.rad;). Although EDTA is commonly included in this assay to eliminate undesirable Fenton side-reactions with H(2)O(2) (a co product of reactions that use xanthine oxidase to produce O(2)(-&z.rad;)), we found that catalase, but not EDTA, blocked suicide elimination of cytochrome c from the reaction. Finally, we demonstrate the feasibility of evaluating superoxide scavenging abilities on small samples extracted from two types of neuronal cultures, a hypothalamic neuronal cell line (GT1 trk cells) and primary mouse cortical cell cultures. This assay allows rapid, high throughput assessments of superoxide scavenging efficacy for small molecules of interest, as well as for cell or tissue extracts. PMID- 10788670 TI - Visualizing and quantifying evoked cortical activity assessed with intrinsic signal imaging. AB - Intrinsic signal imaging (ISI) measures changes in light reflectance from the illuminated cortex (intrinsic signals or IS) attributed to various vascular and metabolic sources that, when using illumination in the 600 nm range, appear to co localize with neuronal activity. Given the multiple sources contributing to the collected IS, the common practice of averaging across an extended post-stimulus time epoch before dividing by baseline data typically visualizes evoked IS overlying both the cortical tissue and the large surface blood vessels. In rat PMBSF, the contribution from these vessels are problematic as they do not co localize with known PMBSF function. Determining a means for quantifying the evoked IS area poses an additional challenge. Here, we describe how exploiting IS collected shortly after stimulus onset (within 1.5 s), which coincides with fast oxygen consumption of active neurons, visualizes evoked IS overlying the cortical tissue without the large surface vessels. We also describe how the use of absolute thresholds combined with a baseline determined from data collected immediately prior to stimulus onset (within 1 s) targets most precisely a specific evoked IS amplitude, a method that should be especially useful when evoked areas are expected to occupy a substantial portion of the total imaged area and/or when peak activity is expected to differ between subjects. PMID- 10788669 TI - Selective measurement of endothelial or smooth muscle [Ca(2+)](i) in pressurized/perfused cerebral arteries with fura-2. AB - Despite a critical role for calcium in endothelial regulation of cerebrovascular tone, endothelial intracellular calcium ([Ca(2+)](i)) has never been measured in the context of an intact pressurized cerebral vessel. The purpose of the present study was to selectively measure endothelial or smooth muscle [Ca(2+)](i) and diameter in a pressurized/perfused cerebral vessel. In a pressurized rat middle cerebral artery, fura-2 AM was administered selectively to either the luminal (endothelium) or abluminal (smooth muscle) side of the vessel. Selectivity of loading was determined by measuring fura-2 fluorescence before and after removal of the endothelium. Removal of the endothelium virtually eliminated fura-2 fluorescence. In addition, 2-methylthioadenosine triphosphate (2MeS-ATP, a selective endothelial P2 receptor agonist) was used to infer the selectivity of fura-2 loading. It was reasoned that 2MeS-ATP should produce a decrease in smooth muscle [Ca(2+)](i) and an increase in endothelial [Ca(2+)](i) in selectively loaded vessels, consistent with its role as an NO-dependent dilator. In smooth muscle loaded vessels, [Ca(2+)](i) went from 252+/-8 to 82+/-9 nM following luminal administration of 2MeS-ATP, whereas in endothelial loaded vessels, [Ca(2+)](i) went from 137+/-11 to 271+/-20 nM. Thus, a method is provided which allows for selective measurement of endothelial or smooth muscle [Ca(2+)](i) with simultaneous measurement of diameter in a pressurized cerebral vessel. PMID- 10788671 TI - Combining laser scanning confocal microscopy and electron microscopy to determine sites of synaptic contact between two identified neurons. AB - Here we report a double labelling method for correlative confocal and electron microscopy (EM) which allows selective characterisation of structural relationships between two single identified neurons in the same preparation. Using the lobster stomatogastric nervous system, we labelled pairs of identified, synaptically-connected neurons by intracellular injection of Lucifer Yellow (LY) in one neuron and a mixture of Rhodamine (Rdh) and Horseradish Peroxidase (HRP) in its partner. First, whole-mounts of LY- and Rdh-stained neurons were visualized using laser scanning confocal microscopy (LSCM) in order to isolate neuropilar regions of possible synaptic contact. Second, after conventional treatment for electron microscopy (LY was revealed with immunogold and HRP with DAB), areas of close appositions were viewed in EM. This technique allowed us to determine all the regions of close contact between two cells, and then to use electron microscopy to determine the presence or absence of synaptic contact within each of these restricted areas. These techniques enabled us to show that there were few areas of apposition and that only an extremely small proportion of these areas was in fact regions of synaptic contact between the two labelled neurons. PMID- 10788672 TI - Mutational analysis of the synapsin III gene on chromosome 22q12-q13 in schizophrenia. AB - Synapsins are a family of neuron-specific, synaptic vesicle-associated phosphoproteins that have been implicated in the modulation of neurotransmitter release. Synapsins are coded by three distinct genes for synapsin I. II, and III. The synapsin III gene is located on human chromosome 22q12-q13, where a possible schizophrenia susceptibility locus is located. Using the single strand conformation polymorphism method, we searched for variants in 13 exons and the 5' flanking region of the synapsin III gene in schizophrenia. Three polymorphisms, 631C/G, -271T/C, and E525Q, and one rare variant, -669C >A, were identified. Case control comparisons of these polymorphisms revealed no significant differences in the allelic and genotypic distributions between schizophrenic and control subjects. The present study did not provide evidence for an association between the synapsin III gene and schizophrenia. PMID- 10788673 TI - Fenfluramine challenge in unipolar depression with and without anger attacks. AB - We have previously hypothesized that patients with major depression and anger attacks may have a greater central serotonergic dysregulation than depressed patients without such attacks. We wanted to compare the prolactin response to fenfluramine challenge, as an indirect measure of central serotonergic function, in depressed patients with and without anger attacks. We recruited 37 outpatients (22 men and 15 women; mean age: 39.5+/-10.5) with DSM-III-R major depressive disorder, diagnosed with the SCID-P. Their initial 17-item Hamilton Rating Scale for Depression score was >/=16. Patients were classified as either having or not having anger attacks with the Anger Attacks Questionnaire. All patients received a single-blind placebo challenge followed by a fenfluramine challenge (60 mg orally) the next day. Plasma prolactin measurements were obtained with double antibody radioimmunoassay before and after both placebo and fenfluramine challenges, and fenfluramine and norfenfluramine blood levels after each challenge were determined by gas chromatography. Of the 37 study participants, 17 (46%) were classified as having anger attacks. There were no significant differences in age, gender, fenfluramine, or norfenfluramine blood levels between depressed patients with and without anger attacks. Depressed patients with anger attacks showed a significantly blunted prolactin response to fenfluramine challenge compared to patients without anger attacks. As previous studies have shown blunted prolactin responses to fenfluramine in impulsive aggression among patients with personality disorders, our results support our hypothesis that depressed patients with anger attacks may have a relatively greater serotonergic dysregulation than depressed patients without these attacks. PMID- 10788674 TI - Relationship between impulsivity and platelet serotonin content in adolescents. AB - The relationship between impulsivity and serotonin function was explored in impulsive and non-depressed adolescents. Platelet serotonin content was chosen as a peripheral indicator of central serotonin function. Impulsivity was assessed with a questionnaire. All measures were performed once a week over a 6-week period for all subjects. Subjects comprised eight adolescent inpatients who were hospitalized as a result of their impulsive acts and eight healthy age- and sex matched control subjects. Mean platelet serotonin concentration was significantly higher in the impulsive group than in the control group. Platelet serotonin concentration was positively correlated with the intensity of impulsivity in the patient group. PMID- 10788675 TI - Mood and energy regulation in seasonal and non-seasonal depression before and after midday treatment with physical exercise or bright light. AB - The effects of two non-drug treatments (physical exercise and bright light) on mood, body weight and oxygen consumption were compared in age-matched groups of female subjects with winter depression, non-seasonal depression or without depression. It was found that oxygen consumption in the pre-treatment condition was similar in non-depressed subjects (n=18) and depressed non-seasonals (n=18), while comparatively lower values were obtained in winter depression (n=27). Neither mood nor metabolic parameters changed significantly in the group of nine untreated winter depressives. One week of physical exercise (1-h pedaling on a bicycle ergometer between 13.00 and 14.00 h) increased oxygen consumption in the group of nine winter depressives and lowered oxygen consumption in nine-subject groups of depressed and non-depressed non-seasonals. One week of bright light treatment (2-h exposure to 2500 lux between 14.00 and 16.00 h) increased oxygen consumption in nine winter depressives and nine non-depressed subjects, while no significant change in oxygen consumption was found in nine subjects with non seasonal depression. Weight loss was observed in the groups treated with physical exercise and in the group of light-treated winter depressives. Winter depression responded equally well to exercising and light, while a significant therapeutic difference in favor of exercising was found in non-seasonal depression. Overall, the results of the study suggest that energy-regulating systems are implicated in the antidepressant action of the non-drug treatments. PMID- 10788676 TI - Increased anti-streptococcal antibodies in patients with Tourette's syndrome. AB - Infection or postinfectious phenomena have been postulated to play a role in the pathogenesis of children afflicted with the typical symptoms of Tourette's syndrome (TS). We investigated whether an increase of titers of antistreptococcal antibodies can be reproduced in our children with TS, and whether this increase is restricted to children. We examined the titers of two different antistreptococcal antibodies, antistreptolysin (ASL) and anitDNase B, both in children and adults. Titer s of ASO and antiDNase B were measured (1) in 13 children/adolescents suffering from TS and in an aged-matched comparison group;(2) in 23 adult patients, a comparison group of 23 aged-matched controls, and in another group of 17 aged-matched, non-medicated acute schizophrenics. ASO and antiDNase B titers were determined by laser nephelometry using a commercially available kit. Two antistreptococcal cut-off levels were compared (> 250 U/ml and 400 U/ml). As expected, increases ASO titers (>400 IU/ml) were found in a higher portion of children/adolescents with TS compared to controls. Regarding adults, titers >250 U/ml for both antistreptococcal antigens were found in significantly more TS patients than in schizophrenic patients or healthy control subjects. The mean values of ASO and antiDNase titers were significantly higher in both groups of TS patients compared to control children/adolescents, to the comparison groups of healthy adults and to schizophrenics. No difference in antistreptococcal titers was found between schizophrenics and the group of healthy adults. TS patients exhibited higher antistreptococcal titers than age-matched comparison groups of both children/adolescents and adults using different types of calculation. Our findings support the theory that a postinfectious immune mechanism may play a role in the pathogenesis of TS. The mechanism still needs to be elucidated. PMID- 10788677 TI - Higher levels of serum copper in schizophrenic patients treated with depot neuroleptics. AB - The findings of previous research on the status of trace elements in patients with schizophrenia have been controversial. We studied 62 outpatients with a DSM IV diagnosis of schizophrenia, and compared them with sex- and age-matched healthy controls. Serum copper levels were significantly higher in schizophrenic patients (mean 117.4 microg/dl; S.D. 23.4) than in healthy controls (105.6+/ 27.9). Those patients on treatment with depot neuroleptics had higher copper levels. Zinc levels did not differ between patients and healthy controls. Altered levels of trace elements in schizophrenic patients may be a consequence of antipsychotic treatment. PMID- 10788678 TI - Mixed anxiety-depressive disorder and major depressive disorder: comparison of the severity of illness and biological variables. AB - Mixed anxiety-depressive disorder (MADD) is a new diagnostic category defining patients who suffer from both anxiety and depressive symptoms of limited and equal intensity accompanied by at least some autonomic features. Patients do not meet the criteria for specific anxiety or depressive disorders. The emergence of the symptoms is independent of stressful life events. There are many issues presently under investigation about the validity of this clinical entity. In this study, a group of 29 patients with MADD was compared with a group of 31 patients with major depressive disorder (MDD) to assess the differences and similarities between these two disease categories in terms of severity measures and biological variables. The dexamethasone suppression test (DST) was employed, and thyroid hormones and thyrotropin (TSH) levels were measured for the evaluation of hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-thyroid (HPT) axes, respectively. The patients with MADD were found to be less depressive and more anxious compared to those with MDD. DST responses and thyroid functions were found to be similar in the two groups. When severity of depression was controlled, k(max) and 2300-h cortisol values were found to be significantly higher in the MADD group. Although the patients with MDD and MADD presented with relatively different clinical features, there is not enough biological evidence indicating that MADD represents a discrete diagnostic category. However, there may be relatively higher HPA activity in MADD patients. PMID- 10788679 TI - Normative data and factor structure of the Temperament and Character Inventory (TCI) in the French version. AB - We explored the psychometric features of the French Temperament and Character Inventory (TCI) in a 602-subject community sample (263 men and 339 women), representative of the French population. The factor structures of the temperament and character dimensions, explored separately, were in agreement with the hypothesized constructs, except for the scales Novelty Seeking NS1 (exploratory excitability), Persistence, and Self-Directedness SD4 (self-acceptance). The internal consistency of the main dimensions was good (Cronbach alpha coefficients between 0.68 and 0.82), but weak for Persistence (0.49). The mean scores of the temperament dimensions were notably different from those published in other normative data - especially lower for Novelty Seeking (16.4+/-5.6) and higher for Harm Avoidance (16.1+/-7.2) when compared with US data - suggesting cross cultural differences in personality assessment, and the necessity to use specific normative values with each translated instrument. PMID- 10788680 TI - Paroxetine binding in aggressive schizophrenic patients. AB - Decreased central serotonergic activity has been associated with aggressive behavior in humans and animals. Whether or not this phenomenon is related to current aggression or to aggressive tendency is debatable. [3H]paroxetine binding in blood platelets represents the activity of serotonin peripheral binding sites. We investigated a possible association between [3H]paroxetine binding in blood platelets and current aggression or homicidal history in schizophrenic patients. Blood platelets of 11 aggressive schizophrenic patients were assayed for [3H]paroxetine binding in blood platelets and compared to findings in 15 non aggressive schizophrenic patients, 15 presently non-aggressive schizophrenic patients with homicidal history, and 15 healthy volunteers. Clinical evaluation was performed using the Positive and Negative Syndrome Scale, the Hamilton Rating Scale for Depression and the Clinical Global Impression scale. B(max) of [3H]paroxetine binding in blood platelets of currently aggressive schizophrenic patients was significantly higher than that in platelets of non-aggressive schizophrenic patients, presently non-aggressive patients with homicidal history and healthy volunteers. No difference was found between the last three study groups. No significant correlation was found between scores of all rating scales and the investigated biochemical parameters. An association was found between current aggression among schizophrenic patients and high B(max) values of [3H]paroxetine binding in blood platelets. This association is probably related to present state of aggression rather than to tendency towards aggression. PMID- 10788681 TI - Evaluation of three simple methods for predicting therapeutic lithium doses. AB - The bias and accuracy of three simple methods for predicting lithium doses were assessed in this prospective study. In each patient, we computed the predicted doses (PDs) of lithium by applying three formulas. The actual dose (AD), the lowest lithium dose that was enough to produce a serum lithium concentration higher than 0.80 mmol/l, was determined. The PD computed by each formula was then compared with the AD to identify the one with the least bias and the greatest accuracy in predicting therapeutic lithium doses. As mean prediction error (ME) is a convenient measure of bias, the prediction error (PE) of each comparison was computed by subtracting the AD from the PD. Accuracy was assessed as a function of the root-mean-squared prediction error (rMSE). Seventeen psychiatric inpatients participated in this study. The 95% confidence interval of ME of a proposed formula [Zetin, M., Garber, D., De Antonio, M., Schlegel, A., Feureisen, S., Fieve, R., Jewett, C., Reus, V., Huey, L.Y., 1986. Prediction of lithium dose: a mathematic alternative to the test-dose method. Journal of Clinical Psychiatry 47, 175-178] was across zero (-15.48 to 133.72). In addition, the rMSE of that formula was also the lowest one (152.66 mg/day). The method proposed by Zetin et al. (1986) is the least biased and the most accurate way to predict therapeutic lithium doses. PMID- 10788682 TI - Chest radiotherapy in limited-stage small cell lung cancer: facts, questions, prospects. AB - OBJECTIVE AND STUDY DESIGN: Limited-disease small cell lung cancer (LD-SCLC) is initially very sensitive to both radiotherapy and chemotherapy. However, the 5 year survival is generally only 10-15%, with most patients failing with therapy refractory relapses, both locally and in distant sites. The addition of chest irradiation to chemotherapy increases the absolute survival by approximately 5%. We reviewed the many controversies regarding optimal timing and irradiation technique. RESULTS: No strong data support total radiation doses over 50 Gy. According to one phase III trial and several retrospective studies, increasing the volume of the radiation fields to the pre-chemotherapy tumour volume instead of the post-chemotherapy volume does not improve local control. CONCLUSIONS: The total time in which the entire combined-modality treatment is delivered may be important. From seven randomized trials, it can be concluded that the timing of the radiotherapy as such is not very important. Some phase III trials support the use of accelerated chest radiation together with cisplatin-etoposide chemotherapy, delivered from the first day of treatment, although no firm conclusions can be drawn from the available data. The best results are reported in studies in which the time from the start of treatment to the end of the radiotherapy was less than 30 days. This has to be taken into consideration when treatment modalities incorporating new chemotherapeutic agents and radiotherapy are considered. PMID- 10788683 TI - Novel approaches to locally advanced unresectable non-small cell lung cancer. AB - The management of advanced non-small cell lung cancer (NSCLC) is rapidly evolving. Advances in combined chemo-radiation therapy have led to improvements in patient survival which are statistically significant, but most patients still succumb to their disease. New chemotherapeutic agents, such as taxanes (paclitaxel, docetaxel), topoisomerase inhibitors (topotecan, irinotecan), and novel analogs (gemcitabine, vinorelbine), may offer the promise of improved outcome, but have not yet been tested in phase III trials. Molecular therapeutics, such as gene therapy, drugs that target specific oncogene activation (such as Ki-ras inactivation by farnesyl transferase inhibitors), and hypoxic cell toxins (such as tirapazamine), are in clinical trials. The optimum use of these agents awaits more rapid and widespread molecular diagnostics. Finally, technological advances in radiotherapy will allow higher tumor doses, while minimizing doses to dose-limiting normal structures, such as the esophagus, normal lung and heart. We describe a move towards molecular strategies, both for therapy and diagnostics, that may result in more effective treatment. While the outcome for patients with advanced non-small cell lung carcinoma is still poor, new agents are being developed rapidly and offer the hope of improved survival. PMID- 10788684 TI - The prognostic impact of quality of life assessed with the EORTC QLQ-C30 in inoperable non-small cell lung carcinoma treated with radiotherapy. AB - PURPOSE: The purpose of this study was to assess the prognostic significance of pretreatment quality of life (QoL) scores and symptom scores in a group of patients treated with high dose radiotherapy. MATERIAL AND METHODS: A total of 198 patients treated with external irradiation (>/=60 Gy) were included. In all these patients, baseline QoL was assessed using the EORTC QLQ-C30. The prognostic significance of a number of non-QoL and QoL parameters with regard to survival was estimated in both univariate and multivariate analyses. RESULTS: In a multivariate model including the non-QoL parameters, performance status, weight loss and N-classification were independent prognostic factors for survival. After entering the QoL parameters in the model, global QoL was the strongest prognostic factor, while performance status lost its significance. Subsequently, a significant interaction term was found between N-classification and global QoL, indicating that global QoL was an independent prognostic factor but that the effect varied as a function of N-status. In N+ patients, the median survival in the group with low scores for global QoL was 4.5 months, which was significantly worse (P<0.0001) compared with the high score group in which the median survival was 12.9 months. CONCLUSION: Global QoL is a strong prognostic factor for survival in patients with NSCLC who have pathological lymph nodes at presentation and who are treated with radical or curative radiotherapy. PMID- 10788685 TI - Effect of CT-based treatment planning on portal field size and outcome in radiation treatment of localized prostate cancer. AB - The portal field sizes of 361 consecutive patients treated with curative radiotherapy for localized prostate cancer were measured. The introduction of CT based information resulted in a significant increase of field sizes, leading to an almost doubling of the treated volume, some increase in late rectal toxicity, but also in local control. PMID- 10788686 TI - An evaluation of three-field coplanar plans for conformal radiotherapy of prostate cancer. AB - BACKGROUND AND PURPOSE: A series of coplanar three-field configurations for two different clinical treatment volumes, prostate only (PO) and prostate plus seminal vesicles (PSV) were studied to determine the optimal three-field plan arrangement for prostate radiotherapy. MATERIALS AND METHODS: A variety of conformal three-field 6 MV plans prescribed to both 64 and 74 Gy were created for PO and PSV volumes in each of ten patients. For description, the orientation of each sequential beam was named in a clockwise fashion. Plans included series with arrangements of 0 degrees, 60-150 degrees, 210-300 degrees; 0 degrees, 90 degrees, 225-255 degrees; 90 degrees, 210-240 degrees, 300-330 degrees and a four field (4F) box plan for comparison. Six-hundred and eighty plans were compared using the rectal volume irradiated to greater than 50% (V(50)), 80% (V(80)), and 90% (V(90)) of the prescribed dose, normal tissue complications (NTCP) for rectum, bladder, and femoral heads (FH), and tumour control probabilities (TCP). FH tolerance was set at 52 Gy to 10% volume. RESULTS: In comparing the 34 different three-field configurations for each of the PO and PSV groups, the greatest rectal sparing was achieved by a three-field plan with gantry angles of 0 degrees, 90 degrees, 270 degrees (PO: rectal V(80)=22.8+/-5.5% (1S.D.), V(90)=18.4+/-5.7%, and PSV: rectal V(80)=41.9+/-5.8%, V(90)=35.5+/-5.9%). This also improved on the 4F-box plan (PO: rectal V(80)=26.0+/-5.8%, V(90)=21.4+/ 5.2%, P<0.001; and PSV: rectal V(80)=47.3+/-5.5%, V(90)=41.6+/-5.1%, P<0.001). The worst rectal sparing was seen with the 0 degrees, 120 degrees, 240 degrees plan (PO: rectal V(80)=35.2+/-8.0%, V(90)=30.3+/-7.1%, P<0.001; and PSV: rectal V(80)=65.7+/-9.0%, V(90)=58.8+/-8.8%, P<0.001). In the PO group, the increase in predicted rectal NTCP with dose escalation from 64 to 74 Gy was 3.3% using the 0 degrees, 90 degrees, 270 degrees plan, 4.7% with the 4F-box plan, and 6.9% with the 0 degrees, 120 degrees, 240 degrees plan. In the PSV group, dose escalation increased the predicted rectal NTCP by 7.9, 10.1 and 15.7% for the 0 degrees, 90 degrees, 270 degrees plan, 4F-box plan, and 0 degrees, 120 degrees, 240 degrees plan, respectively. CONCLUSIONS: For both PO and PSV volumes, the three-field plan which afforded the greatest rectal sparing with acceptable bladder and femoral head doses was the 0 degrees, 90 degrees, 270 degrees plan. This plan also improved on the 4F-box. The increase in predicted rectal NTCP when escalating dose from 64 to 74 Gy was smaller using this plan compared to either the three-field 0 degrees, 120 degrees, 240 degrees plan or the 4F-box plan. PMID- 10788688 TI - Conformal irradiation of concave-shaped PTVs in the treatment of prostate cancer by simple 1D intensity-modulated beams. AB - BACKGROUND: In the case of concave-shaped PTVs including prostate (P) and seminal vesicles (SV), intensity-modulated radiation therapy (IMRT) should improve the therapeutic ratio of the treatment of prostate cancer. PURPOSE: Comparing IMRT by simple 1D modulations with conventional 3D conformal therapy (i.e. non-IMRT) in the treatment of concave-shaped PTVs including P+SV. MATERIALS AND METHODS: For five patients having a concave-shaped PTV (P+SV) previously treated at our Institute with conformal radiotherapy, conventional 3- and 4-fields conformal plans were compared with IMRT plans in terms of biological indices. IMRT plans were generated by using five equi-spaced beams with a partial shielding of the rectum obtainable with our single-absorber modulation technique (Fiorino C, Lev A, Fusca M, Cattaneo GM, Rudello F, Calandrino R. Dynamic beam modulation by using a single dynamic absorber. Phys. Med. Biol. 1995;40:221-240). The modulation was one-dimensional and the shape of the beams was at single minimum in correspondence with the 'core' of the rectum; the beam intensity in the minimum was set equal to 20 or 40% of the open beam intensity. All plans were simulated on the CADPLAN TPS using a pencil-beam based algorithm (with 18 MV X rays). Tumour control probability (TCP) and normal tissue complication probabilities (NTCPs) (for rectum, bladder and femoral head) were calculated for all situations when varying the isocentre dose from 60 to 90 Gy. Dose distributions were corrected taking dose fractionation into account through the linear-quadratic model; for the TCP/NTCP estimations the Webb-Nahum and the Lyman Kutcher models were respectively applied. Three different scores were considered: (a) increase of TCP while keeping rectum NTCP equal to 5% (TCP(5%)); (b) increase of the uncomplicated tumour control probability (P+); (c) increase of the biological-based scoring function (S+), developed by Mohan et al. (Mohan R, Mageras GS, Baldwin B, Clinically relevant optimization of 3D conformal treatments. Med. Phys. 1992;19:933-944). The impact of the uncertainty in the knowledge of the parameters of the biological models was investigated for TCP(5%). RESULTS: (a) The average gain in TCP(5%) when considering IMRT against non-IMRT conformal plans was 7.3% (range 5.0-13.5%); (b) the average increase of P+ was 3.4% (range: 1. 0-8.5%); and (c) the average increase of S+ was 5.4% (range 2.9-12. 4%). The largest gain was found for one patient (patient 5) showing a significantly larger overlapping between PTV and rectum. CONCLUSIONS: Simple 1D-IMRT may clearly improve the therapeutic ratio in the treatment of concave-shaped PTVs including P and SV. In the range of clinically suitable values, the impact of the uncertainty of the parameters n and sigma(alpha) does not significantly alter the main results concerning the gain in TCP(5%). The reported gain in terms of P+ and S+ should be considered with great caution because of the intrinsic uncertainties of the model's parameters and, for bladder, because the 'true' DVH (considering variations of the shape and dimension due to variable filling) may be very different from the DVH calculated on a single CT scan. Further investigations should consider inversely-optimised 1D and 2D-IMRT plan in order to compare them in terms of cost-benefit. PMID- 10788687 TI - Feasibility study combining low dose rate (192)Ir brachytherapy and external beam radiotherapy aiming at delivering 80-85 Gy to prostatic adenocarcinoma. AB - BACKGROUND: Increasing the radiation dose to prostatic adenocarcinoma has provided higher local control rates. A total of 80 Gy seem necessary to achieve this goal but patient set-up and prostate motion remain difficult problems to solve in conformal radiotherapy. Brachytherapy which overcomes these points could be an alternative way to external beam boost fields. We wanted to transpose the irradiation models largely used in cervix cancer treatment combining external beam radiotherapy and low dose rate brachytherapy. MATERIALS AND METHODS: In 71 patients with 19.5 and 13 ng/ml mean and median PSA levels, respectively, a dose escalation from 74 to 85 Gy was performed in four groups. RESULTS: Shifting from intraoperative placement of sources vectors (Group I) to positioning under ultrasound controls (groups II-IV), improving the implantation shape and optimizing radiation delivery to urethral bed have reduced the total dose to rectal wall under 65 Gy and to urethra under 100 Gy. Rectal/prostate dose ratio was lowered from 0.7 (Groups I-II) to 0.58 (Groups III-IV) while avoiding problems resulting from pelvic bone arch interference, prostate volume or seminal vesicles location. The mean and median follow-up periods are 28 and 18 months. In Groups III and IV 85% of patients without hormonotherapy treated with 80-85 Gy normalized PSA under 1 ng/ml within 6 months. No severe late effect has been noted for patients implanted under echographic control. CONCLUSIONS: The method described allows to deliver 85 Gy. Longer follow-up is however needed but the levels of dose delivered are not expected to induce prohibitive side effects. PMID- 10788689 TI - Seed misplacement and stabilizing needles in transperineal permanent prostate implants. AB - BACKGROUND AND PURPOSE: Seed misplacement occurring in transperineal permanent implants contributes to the degradation in dose coverage. It has been suggested that needles could be used to immobilize the prostate and help reduce misplacement. This study investigates the effects of parallel stabilizing needles on seed misplacement. MATERIALS AND METHODS: A group of ten patients implanted with stabilizing needles was compared with a group of 20 patients implanted without stabilization. Measurements were performed on the displacement of individual seeds and needles. The needle measurements are: insertion angle, the ratio of post-implant over pre-implant lengths and the clustering tendency, a measure of relative misplacement among the seeds of the same needle. RESULTS: No difference was observed in seed misplacement. No difference was observed in needle insertion angle, a measure which was expected to improve with the use of stabilizing needles. CONCLUSION: None of the expected effects from the use of parallel stabilizing needles have been observed. This method of prostate contention appears to be without benefits. Seed misplacement is most pronounced along the insertion axis and is caused by friction between prostatic tissues and implantation needles. Reducing friction could be a promising alternative to prostate contention in trying to reduce misplacement. PMID- 10788690 TI - Applications of simulator computed tomography number for photon dose calculations during radiotherapy treatment planning. AB - PURPOSE: To study the variation of computed tomography (CT) number from a simulator-based scanner and the effect of this variation on photon-dose calculations. METHOD AND MATERIALS: CT images of a cylindrical phantom with multiple inserts were obtained using a commercially-available simulator-CT (Ximatron: Varian, Palo Alto, CA). The linear correlation coefficient and Chi square methods were used to determine the X-ray effective energy in a phantom. CT numbers in Hounsfield units (HU) were measured as a function of phantom size, orientation, field of view (FOV), distance from the center, and time for various inserts. The change of dose calculations due to the CT number variations was then determined using the equivalent path-length (EPL) and collapsed cone convolution methods. RESULTS AND DISCUSSION: A significant beam-hardening effect was observed for the simulator-CT. Consequently, the CT number from the sim-CT was more sensitive to the size of the phantom than those from a conventional CT. The sim CT number is not sensitive to the locations within the phantom and is stable over a 6-week period. It is important to use the proper FOV for sim-CT studies; scanning a small polystyrene phantom using a large FOV may result in an increase of l20 HU in CT number at the center of the field. However, the dose-calculation variations, due to the CT number uncertainty, do not exceed 2-3% for 6-18 MV photon beams. CONCLUSION: The simulator CT images were acquired with patients in the treatment position, and these CT numbers are useful for CT-based dose calculations. PMID- 10788692 TI - Curved angiocatheter metal guide for the implantation in velo-tonsillar carcinomas. AB - Brachytherapy in velo-tonsillar region is not applied in all the institutions because of difficulties to learn it. For the implantation of wires in the soft palate, the Reverdin needle is replaced by a curved guide of angiocatheter 14 Gauge. This technique is easy and reduces the risk of tear mucosa. PMID- 10788691 TI - A CT-assisted method of dosimetry in brachytherapy of lung cancer. Rotterdam Oncological Thoracic Study Group. AB - BACKGROUND: The toxicity of endobronchial brachytherapy (EB), in particular fatal haemoptysis and bronchial wall necrosis, has been correlated with the total dose, fraction size, volume encompassed by the 100% isodose, and a proximal tumor location. We describe a CT-based planning method which, by improving target volume definition and volumetric dose information, can improve the therapeutic ratio of EB. MATERIALS AND METHODS: Sixteen CT-assisted EB procedures were performed in patients who were treated with palliative high-dose rate EB. The CT data were used to analyze applicator position in relation to anatomy. An example of a three-dimensional optimized treatment plan was generated and analyzed using different types of dose-volume histograms. RESULTS: The procedure was well tolerated by patients and no post-procedure complications were observed. The bronchial applicator was eccentrically positioned at the level of the carina/mainstem bronchus in 12 (of 14) CT scans. A planning CT prior to EB was not found to be useful as the final target volume and/or the final applicator position were not reliably predicted before the therapeutic bronchoscopy. CT scans performed with the applicator in situ allowed the bronchial segments in the target volume to be identified and enabled dose prescription to the bronchial mucosa. CONCLUSIONS: CT-assisted EB is feasible and underlines the need for using centered applicators for proximally located tumors. By enabling accurate mucosal dose prescription, CT-assisted EB may reduce the toxicity of fractionated EB in the curative setting. However, faster on-line EB treatment planning is needed for the routine clinical application of this technique. PMID- 10788693 TI - Mechanisms underlying the M-current block by barium in bullfrog sympathetic neurons. AB - Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the channel blocking actions of barium (3-2000 microM) on an M-type potassium current (I(M)). Barium (IC(50) approximately 105 microM) blocked I(M) without affecting the 50%-activation voltage ( approximately -35 mV) and the slope factor ( approximately 11 mV) of the activation curve. The results indicate that the barium block is independent of the kinetics of I(M). PMID- 10788694 TI - Reduced nitric oxide is involved in prenatal ischemia-induced tolerance to neonatal hypoxic-ischemic brain injury in rats. AB - To explore the role of nitric oxide (NO) in the hypoxic-ischemic (HI) tolerance phenomenon, NO production and brain injury following neonatal hypoxia-ischemia (induced by unilateral common carotid artery ligation followed by hypoxic exposure) were assessed in rat pups with or without HI preconditioning. A previously demonstrated prenatal HI rat model of preconditioning was used in this study. On G17, rat fetuses were subjected to either HI in utero (PreHI) for 30 min or a sham operation (SH). The PreHI treatment provided significant protection against neonatal HI-induced brain injury, as indicated by decreased ipsilateral brain weight reduction, less severe tissue damage, and decreased activation of caspase-3. Concomitant with the protective effect of prenatal HI preconditioning, elevation of nitrite/nitrate content in the ipsilateral cortex of the brain, as an indirect measure of NO production, was significantly lower in the PreHI group than in the SH group following neonatal HI. The protective effect of prenatal HI preconditioning could be reversed by sodium nitroprusside (SNP), a spontaneous NO donor, while SNP had no effect on neonatal HI-induced brain injury in the SH group. Intraperitoneal administration of SNP to pups from the PreHI group (2 mg/kg, 24 and 1.5 h before neonatal HI) increased neonatal HI-induced brain injury similar to that observed in the SH group. On the other hand, L-N(G)-nitro arginine (2 mg/kg, i.p., 1.5 h before the hypoxic exposure), an NO synthase inhibitor, significantly attenuated neonatal HI-induced brain injury in the SH group. The overall results indicate that reduced NO production in the preconditioned rat brain contributes to prenatal HI-induced tolerance to neonatal HI brain injury. PMID- 10788695 TI - Calretinin is present in serotonin- and gamma-aminobutyric acid-positive amacrine cell populations in the retina of Xenopus laevis. AB - In the frog retina most bipolar cells, sparsely distributed amacrine cells and some ganglion cells contain calretinin (CaR). Double-label immunocytochemistry shows that in the Xenopus retina many calretinin positive amacrine cells are also gamma-aminobutyric acid (GABA)-immunoreactive (IR), none colocalizes glycine or dopamine but some contain serotonin (SER). The CaR-IR cells represent 8-9% of all GABA-IR amacrine cells. Only 4.6% of the CaR-positive cells contain SER. The SER positive cells are present in two sizes in the anuran retina: the large cells never contain CaR but some of the small cells do. To further characterize the CaR+/SER+ amacrine cell population, immunolabeling for tryptophan hydroxylase (TrOH), the rate-limiting enzyme for SER synthesis, was performed. The results showed that large TrOH-containing cells are always negative for CaR. However, the small SER-synthesizing amacrine cells are invariably CaR-positive. Thus the anuran retina contains three neurochemically distinct SER-positive amacrine cell types, one of which (the small SER-synthesizing cell type) is also CaR-IR. PMID- 10788696 TI - Fine structural evidence of mechanoreception in spinal lumbosacral accessory lobes of pigeons. AB - In the lumbosacral spinal cord of birds there are accessory lobes which protrude into the vertebral canal. The accessory lobes consist of multipolar neurons and glia-derived glycogen cells. It has been suggested that these lobes function as a sense organ of equilibrium. Therefore the lobes were studied ultrastructurally to look for possible mechanoreceptive structures. Extracellular lacunae extend from the periphery deep into each lobe. The dendrites of neurons ramify into the lacunae where they issue finger-like processes which do not contact any other cells and which are not contacted by boutons. Since finger-like processes are typical of peripheral and central mechanoreceptive neurons it is concluded that the observed processes indicate a mechanoreceptive function of the lobe neurons. PMID- 10788698 TI - Patients with somatoform phobic postural vertigo: the more difficult the balance task, the better the balance performance. AB - The objective was to test, whether the increased body sway activity, shown in patients with phobic postural vertigo (PPV) in a previous posturographic study, impairs postural balance during demanding balance tasks. In 17 patients with PPV and 15 normal subjects body sway was analyzed for two standing positions on a foam rubber-padded posturographic platform with the eyes open or closed: (a) normal upright stance, (b) tandem stance. During normal upright stance patients showed an increase in body sway activity between 0.1 and 19 Hz and in sway path values for lateral and fore/aft directions. During the most difficult balance task, i.e. tandem stance with the eyes closed, body sway activity and sway path values did not differ between patients and controls. Objective balance skills were not impaired in patients with PPV during balance tasks at the limits of postural control. PMID- 10788697 TI - Influence of muscarinic ligands on the amplitudes of the evoked surface potential's late components in the optic tectum of the urodele Plethodon jordani. AB - Recordings of field potentials from the tectal surface of an urodele amphibian were obtained in an in vitro preparation under influence of various muscarinic drugs. Bath applied acetylcholine (ACh) led to no change in the amplitudes or the shape of the evoked potentials. If the ACh-esterase blocker (-)-physostigmine was applied synchronously, the late components of the surface potential increased in amplitude. The non-selective cholinergic agonist carbachol showed a similar effect which was partially diminished by the nicotinic antagonist d-tubocurarine chloride (d-TC) and the muscarinic antagonist atropine sulfate. The application of the non-selective muscarinic agonist (+)-pilocarpine hydrochloride led to an increase of the late oligo- and polysynaptic events. This effect was reduced by the M(1)-antagonist pirenzepine dihydrochloride. The presented findings suggest that muscarinic receptors play a more important role in tectal processing than assumed in previous studies which emphasized the role of nicotinic receptors. PMID- 10788699 TI - Spatial-temporal response characteristics of the ON-OFF direction selective ganglion cells in the rabbit retina. AB - We studied the response properties of the ON-OFF direction selective (DS) ganglion cells in the rabbit retina. The activities of the ganglion cells were recorded extracellularly with tungsten-in-glass electrodes. Drifting sine-wave gratings with various spatial and temporal frequencies were used to stimulate the ON-OFF DS cells. Individual ON-OFF DS cell yielded strongest response to a particular spatial frequency irrespective of temporal frequency. The optimal spatial wavelength increased with increasing eccentricity and its half-width approximately matched the size of the receptive field. The optimal spatial frequency implies that the directional inhibition is effective over a range of about a receptive field diameter. The characteristics of the ON-OFF DS cells suggest that they might function as local motion detectors. PMID- 10788700 TI - Molecular determinants of binding of a wasp toxin (PMTXs) and its analogs in the Na+ channels proteins. AB - The structural specificity of alpha-PMTX, a novel peptide toxin derived from wasp venom has been studied on the neuromuscular synapse in the walking leg of the lobster. alpha-PMTX is known to induce repetitive action potentials in the presynaptic axon due to sodium channel inactivation. We synthesized 29 analogs of alpha-PMTX by substituting one or two amino acids and compared threshold concentrations of these mutant toxins for inducing repetitive action potentials. In 13 amino acid residues of alpha-PMTX, Arg-1, Lys-3 and Lys-12 regulate the toxic activity because substitution of these basic amino acid residues with other amino acid residues greatly changed the potency. Determining the structure activity relationships of PMTXs will help clarifying the molecular mechanism of sodium channel inactivation. PMID- 10788701 TI - Expression of alpha-calcitonin gene-related peptide in the enteric nervous system of rat small intestine. AB - We first detected alpha-calcitonin gene-related peptide (alpha-CGRP) precursor mRNA in the enteric nervous system (ENS) of rat small intestine by reverse transcriptase polymerase chain reaction (RT-PCR). The nucleotide sequence of the RT-PCR product was completely identical to that found in other organs. By in situ hybridization using digoxygenin-labeled alpha-CGRP precursor cRNA probe, we found that antisense probes detected a signal on nerve cell bodies of both submucosal and myenteric plexuses. Our findings indicate that the rat ENS participates in synthesis of alpha-CGRP precursor. PMID- 10788702 TI - Effect of electroacupuncture on blood pressure and adrenal nerve activity in anesthetized rats. AB - The neural mechanism underlying the effect of electroacupuncture (Ea) on arterial blood pressure (BP) and adrenal nerve activity (ANA) was investigated in anesthetized rats. Tsusanli (St-36) and Hoku (Li-4) were tested with combinations of two different frequencies (3 and 30 Hz) with various stimulation intensities of Ea. At Tsusanli, no effect was found, while at Hoku, an elevation of BP in parallel with ANA was elicited during Ea when the intensity was 5xT or higher. The pattern of the pressor response caused by the low frequency Ea (LFEa, 3 Hz) was a tonic one, while a phasic one was induced by the high frequency Ea (HFEa, 30 Hz). When both Hoku were simultaneously stimulated with the same frequency, the latency to reach the maximal effect was shortened. However, when two different frequencies were used instead, a response characterized by a combination of both phasic and tonic effect was obtained. In bilateral Ea with idential frequency but different onset time, the pressor effect elicited by the latter Ea showed no further increase during the stimulation period, however, when different frequencies were employed, each Ea elicited its own effect independently. The pressor effect elicited by Ea was abolished by regitine but not affected by adrenalectomy. It is concluded that a LFEa and a HFEa at Hoku with appropriate stimulation parameters can increase BP which is mainly due to potentiation of the sympathetic vasoconstrictor tone but via different central mechanisms. PMID- 10788703 TI - Sensory representation of passive movement kinematics by rat's spinocerebellar Purkinje cells. AB - In this paper we examined Purkinje cells' sensory representations of kinematic parameters of passive movements imposed to the forelimb of anesthetized rats. Simple spike Purkinje cell activity was recorded while the rat's ipsilateral forearm was moved passively along circular footpaths at two different speeds. We found that the activity of 35.33% (165/467) of the neurons was significantly modulated during movement cycles. A multivariate regression analysis indicated that movement direction was the predominant factor in determining Purkinje cell activity, whereas movement velocity (i.e. the combination of movement direction and speed) was represented to a much lesser degree. Based on this result, we might suggest that a cortical efferent copy is necessary to the cerebellum in order to elaborate a movement velocity signal. PMID- 10788704 TI - The genesis of human event-related responses explained through the theory of oscillatory neural assemblies. AB - The goal of the study is to investigate the contribution of delta and theta responses to N200 and P300 ERP components that are recorded from two topographical sites (Fz and Pz) under two experimental paradigms (mismatch negativity and oddball) that trigger different cognitive processes for the respective task performances. The present study was conducted on 42 normal young adults. The results showed that it is the 'interplay' between the theta and the delta oscillations that produces the morphology and the amplitude not only of the P300 but also the N200 component. The functional and physiological meaning of the delta and theta responses are discussed within the framework of the theory of oscillatory neural assemblies and the principle of superposition. PMID- 10788705 TI - Quantification of tau phosphorylated at threonine 181 in human cerebrospinal fluid: a sandwich ELISA with a synthetic phosphopeptide for standardization. AB - Hyperphosphorylation of the microtubule-associated protein tau is specifically found in those brain cells affected in several tauopathies. Tau has also been consistently found to be present in the cerebrospinal fluid (CSF). Here we report the quantification in CSF of tau phosphorylated at Thr 181 using an immunoassay with a synthetic peptide for standardization. The choice of the peptide was based on fine mapping of a phospho-dependent antibody, AT270 (P(176)PAPKT(p)P(132))and a human specific tau antibody, HT7 (P(159)PGQK(163)). CSF-phospho-tau levels were increased in Alzheimer patients (23.5+/-10.1 pM, P<0.01) compared with age matched controls (15.9+/-5.7 pM), while decreased in patients with frontotemporal dementia (8.6+/-3.9 pM; P<0.01). In every diagnostic group, a highly significant correlation was found between total tau and phospho-tau (Alzheimer's disease, r(2)=0.73; frontotemporal dementia, r(2)=0.43; Control, r(2)=0.42), suggesting that the degree of phosphorylation of CSF-tau changes in different clinical conditions. PMID- 10788706 TI - Functional inactivation of dorsal hippocampus impairs active place avoidance in rats. AB - This paper assesses the contribution of hippocampus to the spatial orientation of Long-Evans rats in a new place avoidance task. The animals learn to avoid a mild footshock in a segment of a rotating arena. Since the punished region is defined in the coordinate system of the stationary room the subject is forced to move away from the prohibited segment even if it is immobile. After bilateral injection of tetrodotoxin (5 ng in 1 microl of saline) into the dorsal hippocampus rats were not able to avoid the punished place while a similar injection of saline did not affect performance. The results suggest the task is suitable for assessing the hippocampus-dependent spatial abilities of laboratory rodents. PMID- 10788707 TI - Developmental changes in expression of the three ryanodine receptor mRNAs in the mouse brain. AB - Ryanodine receptors (RyR) are Ca(2+)-induced Ca(2+) release channels located on the endoplasmic reticulum, and consist of three isoforms, termed RyR1-3. We examined their expression in developing mouse brains by in situ hybridization. During the embryonic stage, RyR1 mRNA levels were highest in the rostral cortical plate, whereas RyR3 mRNA was most prominent in the caudal cortical plate and hippocampus. Initially, low levels of RyR2 mRNA were distributed in the diencephalon and brainstem. However, from postnatal day 7 onward, RyR2 mRNA became the major isoform in many brain regions, while RyR1 mRNA became prominent in the dentate gyrus and Purkinje cell layer. Postnatal down-regulation in the caudal cerebral cortex restricted RyR3 mRNA expression to the hippocampus, particularly the CA1 region. Therefore, RyR expression undergoes dynamic changes during the early postnatal period, when neurons are undergoing structural and functional differentiation. PMID- 10788708 TI - Dysfunction of rat forebrain astrocytes in culture alters cytokine and neurotrophic factor release. AB - Altered glial cell function occurring in substantia nigra in Parkinson' disease may lead to the release of cytokines and impairment of neurotrophic factor production, which in turn, may cause dopaminergic apoptosis. To evaluate this concept, primary cultures of rat brain astrocytes were activated with lipopolysaccharide (LPS), depleted of glutathione with L-buthionine-[S,R] sulfoximine or subjected to complex I inhibition with 1-methyl-4 phenylpyridinium. The effects on tumour necrosis factor-alpha (TNF-alpha) release, dopamine-stimulated glial cell line derived neurotrophic factor (GDNF) and brain derived neurotrophic factor (BDNF) release were determined. LPS activation or inhibition complex I activity, but not glutathione depletion, stimulated TNF-alpha release. Glutathione depletion or complex I inhibition, but not LPS-induced activation, impaired dopamine-stimulated GDNF release. None of these treatments altered BDNF release. Thus, altered glial function leading to TNF-alpha-mediated or GDNF withdrawal-induced dopaminergic apoptosis may contribute to nigral degeneration in Parkinson's disease. PMID- 10788709 TI - Interleukin-2 gene deletion produces a robust reduction in susceptibility to experimental autoimmune encephalomyelitis in C57BL/6 mice. AB - Dysregulation of interleukin-2 (IL-2), the prototypical T cell growth factor and immunoregulatory cytokine, may modify self-tolerance and predisposition to autoimmunity. The available literature suggested that IL-2 could be hypothesized to either propagate or inhibit the development autoimmune demyelinating disorders of the central nervous system such as multiple sclerosis. Thus, the present study sought to test these competing hypotheses by examining whether disrupting one or both IL-2 gene alleles would render mice more or less vulnerable to experimental autoimmune encephalomyelitis (EAE). Myelin oligodendrocyte glycoprotein was used to induce EAE in C57BL/6-IL-2(-/-) knockout, C57BL/6-IL-2(+/-) heterozygote and C57BL/6-IL-2(+/+) wild-type mice. All of the wild-type and heterozygote mice developed signs of EAE compared with only 23% of the IL-2 knockout mice. Histopathological examination of lumbar spinal cord sections confirmed that subpial perivascular inflammatory infiltrates found in wild-type and heterozygote mice were absent in the unaffected IL-2 knockout mice. These data demonstrate that vulnerability to EAE is markedly reduced in C57BL/6 mice lacking IL-2, and suggest that this cytokine may play a critical role in autoimmune processes of the central nervous system. PMID- 10788710 TI - Blockade of pilocarpine-induced cerebellar phosphoinositide hydrolysis with metabotropic glutamate antagonists: evidence for an indirect control of granule cell glutamate release by muscarinic agonists. AB - The ability in vivo of the muscarinic agonist, pilocarpine, to increase phosphoinositol (PI) hydrolysis in lithium pretreated rats was investigated by measuring the accumulation of [(3)H]inositol phosphates (IP). As expected, 20 mg/kg s.c. pilocarpine, a muscarinic agonist, increased PI hydrolysis in the striatum, frontal cortex and hippocampus. Somewhat surprisingly, an increase in IP was also found in the cerebellar homogenates. In all four tissues the pilocarpine-induced effect could be completely inhibited by pretreatment with the muscarinic antagonist scopolamine (1.2 mg/kg i. p.). It was also found that the cerebellar but not the hippocampal pilocarpine-induced rise in PI hydrolysis could be blocked by the metabotropic glutamate (mGlu) receptor antagonist, LY341495 (100 nmol, i.c.v.). The same dose of LY341495 was found to also block both the cerebellar and hippocampal increase in IP formed by stimulation with the group I mGlu receptor agonist 3, 5-dihydroxyphenylglycine (1 micromol, i.c.v.). Given this data and the current information on the distribution of muscarinic and mGlu receptors in the cerebellum, it is suggested that these results may be a reflection of pilocarpine acting at M(2) receptors to indirectly increase glutamate release from parallel fibers by inhibition of gamma-aminobutyric acid releasing Golgi cells. PMID- 10788711 TI - Functional synapse formation between rat olfactory receptor neurons and olfactory bulb neurons in vitro. AB - To explore the mechanism of the synapse formation between olfactory receptor neurons and neurons in the olfactory bulb, blocks of olfactory epithelium and slices of olfactory bulb were cocultured by a modified slice culture method. After 4 days in culture, neuron specific fibers from the olfactory epithelium block extending to the olfactory bulb slice were observed. After approximately 20 days in culture, application of forskolin, which induces excitatory responses in olfactory receptor neurons, to the epithelium induced inward current responses in olfactory bulb neurons under the voltage clamp conditions, indicating that functional synapses between olfactory receptor neurons and olfactory bulb neurons had been established in vitro conditions. PMID- 10788712 TI - More than meets the eye: significant regional heterogeneity in human cortical T1. AB - Segmented k-space acquisition of data was used to decrease the acquisition time and to increase the imaging resolution of the precise and accurate inversion recovery (PAIR) method of measuring T(1). We validated the new TurboPAIR method by measuring T(1) in 158 regions of interest in 12 volunteers, using both PAIR and TurboPAIR. We found a 3% difference between methods, which could be corrected by linear regression. After validation, the TurboPAIR method was used to test a hypothesis that there is significant regional heterogeneity in cortical T(1). We measured cortical gray matter T(1) in 11 right-handed volunteers, in 48 regions of interest scattered over frontal and parietal cortex, and in 46 ROIs along the central sulcus (CS). We found that T(1) in the CS is less than T(1) elsewhere in the cortex (p<0.001), and that there is considerable hemispheric asymmetry in T(1) in gray matter, but not in white matter. In central gray structures (caudate, thalamus, nucleus pulvinarus), and in the posterior CS (sensory cortex), right hemisphere T(1) was significantly greater than left hemisphere T(1) (p< or =0.004). In cortical gray matter of the frontal lobe and anterior CS (motor cortex), left hemisphere T(1) was significantly greater than right hemisphere T(1) (p< or =0.003). These findings demonstrate that there is considerable regional heterogeneity in human cortical T(1) that is unexplained by differences in tissue iron content, but may be evidence of an inherent anatomic asymmetry of the brain. PMID- 10788713 TI - Correlating magnetic resonance imaging markers of axonal injury and demyelination in motor impairment secondary to stroke and multiple sclerosis. AB - The primary pathological mechanisms in stroke and multiple sclerosis (MS) are very different but in both diseases, impairment may arise from a final common pathway of axonal damage. We aimed to examine the relationship between motor impairment, magnetisation transfer ratio (MTR) (an index of demyelination), and N acetyl aspartate (NAA) loss (an index of axonal injury) localised to the descending motor pathways in stroke and MS. Twelve patients between 1 and 10 months after first ischaemic stroke causing a motor deficit and 12 patients with stable MS with asymmetric motor deficit were examined. T(2)-weighted imaging of the brain together with MTR and proton (voxel 1.5x2x2 cm(3)) MRS localised to the posterior limb of the internal capsule were performed and correlated to a composite motor deficit score. MTR and NAA in the internal capsule were reduced in both stroke and MS patients compared to controls. NAA loss correlated with motor deficit score in both stroke and MS (p<0.001 and p = 0.04, respectively). Correlations were seen between MTR and motor deficit (p<0.001) MTR and NAA loss (p <0.001) in stroke patients but not in MS patients. Axonal injury in the descending motor tracts would appear to be an important determinant of motor impairment in both stroke and MS. In stroke, MTR measures of demyelination are closely related to axonal damage and thus also correlate with motor deficit. However in MS, MTR measures of demyelination do not correlate with NAA loss or motor deficit suggesting that demyelination and gliosis may occur independently of axonal damage and are less closely linked with functional impairment. PMID- 10788714 TI - Saturation transfer ratio imaging in invasive ductal carcinomas of the breast. AB - A prospective study was performed to investigate the correlations between saturation transfer ratio (STR) and histologic parameters of invasive ductal carcinomas in human breast. The histologic parameters investigated were the extent of fibrosis in the intercellular matrix, dysplastic changes of nuclei, and mitotic index. Twenty-seven patients with breast carcinoma were examined using an off-resonance saturation pulse in conjunction with conventional field-echo T(1) weighted imaging at frequency offsets of 448 Hz and 1200 Hz from water resonance. The values of STR at frequency offset of 1200 Hz (STR(1200)) increased from non scirrhous carcinoma to scirrhous carcinoma. Although STR(1200) showed correlation with the extent of fibrosis in the intercellular matrix (p<0.01, n = 27), they did not correlate with the dysplastic changes of nuclei or mitotic index. On the other hand, the values of STR at frequency offset of 448 Hz (STR(448)) demonstrated close correlation to dysplastic changes of nuclei and mitotic index (p<0.01, n = 27). STR(1200) correlates with the structural characteristics and STR(448) correlates with the nature of malignant cells with regard to nuclear dysplasia and mitotic potential. PMID- 10788715 TI - Origin of subarachnoid cerebrospinal fluid pulsations: a phase-contrast MR analysis. AB - Cerebrospinal fluid (CSF) pulsations result from change of blood volume in the closed craniospinal cavity. We used cine phase contrast MR analysis to determine whether spinal CSF pulsations result from spinal vascular pulsations or intracranial subarachnoid pulsations, whether intracranial CSF pulsations result from intracranial large arteries pulsations or cerebrovascular bed changes. We performed a quantified physiological mapping of CSF velocity waveforms along the craniospinal axis. Thirty-six volunteers participated in the study. MR acquisitions were obtained at the intracranial level, the upper, midcervical, cervicothoracic, mid thoracic, and/or the thoracolumbar levels. The temporal velocity information were plotted as wave form and key temporal parameters were determined and analyzed; intervals from the R wave to the onset of CSF systole, to CSF systolic peak, to the end of systole, as well as duration of systole. Three kinds of dynamic channels could be differentiated along the spinal axis, the lateral, medioventral and mediodorsal channels. Lateral spinal CSF pulse waves show significant craniocaudal propagation. No such significant progression was detected through the medial channels along the spine. Through the medial channels, a cephalic progression was observed from the upper cervical level to the intracranial level. At the craniocervical junction, mediodorsal CSF systole appeared the earliest one whereas in the anterior intracranial basal cistern, CSF systole appeared delayed. In conclusion, spinal CSF pulsations seem to result mainly from intracranial pulsations in the lateral channels, whereas local vascular pulsations could modify CSF pulse wave mainly in the medial channels. At the craniocervical junction, our results suggest that blood volume change in the richly vascularised cerebellar tonsils is the main initiating factor of CSF systole; and that spinal vascular pulsations could be considered as an additional early and variable CSF pump. PMID- 10788716 TI - Analysis of activation in anterior cingulate cortex during cognitive process of selection following somatosensory stimuli: fMRI study with elaborate task paradigms. AB - The anterior cingulate cortex (ACC) has been pointed out to play an important role in the cognitive process of selection underlying "early selection" of perceptually (visually or auditorily) and "late selection" of behaviorally relevant information. However, it is still unclear in cognitive process of selection that the ACC can be activated by somatosensory stimuli as perceptually relevant information. To determine whether the ACC is activated by "early selection" of somatosensory stimuli surely without effects of motor acts as behavior, eighteen normal subjects performed elaborately designed tasks of selection while receiving somatosensory stimuli on their toes of the right and left feet under three different conditions using functional magnetic resonance imaging (fMRI) at 1.5 T. ACC activation was observed to be 2.1+/-0.3% (mean +/- SE) in selection and finger movement as motor acts, and 1.3+/-0.3% in selection and counting (without motor acts), whereas there was no activation in non selection. The present fMRI study demonstrates that the ACC is activated by "early selection" following somatosensory stimuli surely without subsequent motor acts. PMID- 10788717 TI - High-resolution segmented EPI in a motor task fMRI study. AB - A high-resolution gradient echo, multi-slice segmented echo planar imaging method was used for functional MRI (fMRI) using a motor task at 1.5 Tesla. Functional images with an in-plane resolution of 1 mm and slice thickness of 4 mm were obtained with good white-gray matter contrast. The multi-shot approach, combined with a short total readout period of 82 ms, limits blurring effects for short T(2)(*) tissues (such as gray matter), assuring truly high-resolution images. In all subjects, motor functions were clearly depicted in the contralateral central sulcus over several slices and sometimes activation was detected in the supplementary motor area and/or ipsilateral central sulcus. The average signal change of 11+/-3% was much higher than in standard low-resolution fMRI EPI experiments, as a result of larger relative blood fractions. PMID- 10788718 TI - Histopathological analysis of a bladder cancer stalk observed on MRI. AB - Papillary transitional cell carcinoma of the bladder has a loose connective tissue stalk. For staging of bladder cancer on magnetic resonance imaging (MRI), it is important to clearly separate the cancer from the bladder wall. It is possible to distinguish a stalk from the cancer by the difference of intensity on the using MRI. Sixteen stalks of 20 polypoid bladder tumors on any of the T(2)W(I), dynamic images and delayed enhanced images were demonstrated. Most of the stalks show lower signal intensity than the tumors on T(2)W(I), less enhancement on dynamic images and stronger enhancement on delayed enhanced images. The stalk consisted of fibrous connective tissue, capillary blood vessels, inflammatory cell infiltration and edema. This stalk extended from the bladder wall to the center of the tumor. Some of the superficial muscular bundles were pulled into the stalk. These histopathological findings were compatible with the patterns of signal intensities on MRI. The identification of the stalk of a polypoid tumor may be an important observation to exclude bladder wall invasion by tumor. PMID- 10788719 TI - Congenitally corrected transposition of the great arteries (L-TGA) with situs inversus totalis in adulthood: findings with magnetic resonance imaging. AB - Diagnosis of congenitally corrected transposition of the great arteries (L-TGA) with situs inversus totalis in two adult patients was made by magnetic resonance imaging (MRI). Visualization of the complete anatomy and quantification of ventricular function was possible. Relevant concomitant disease such as perimembraneous ventricular septal defect, atrial secundum septal defect, tricuspid regurgitation, valvular pulmonic stenosis, and pulmonary artery dilatation were clearly depicted by MRI using standard spin-echo and gradient echo techniques. Findings were confirmed by cardiac catheterization in both patients. In this rare and complex congenital cardiac anomaly, MRI is an excellent imaging modality as echocardiography may be difficult to interpret due to restricted imaging windows. MRI may help in the decision about the necessity to undergo further invasive evaluation and may help to make cardiac catheterization a straightforward procedure. PMID- 10788720 TI - Evaluation of water content by spatially resolved transverse relaxation times of human articular cartilage. AB - Non-invasive assessment of cartilage properties, specifically water content, could prove helpful in the diagnosis of early degenerative joint diseases. Transverse relaxation times T(2) of human articular cartilage (34 cartilage slices of three donors) were measured on a pixel-by-pixel basis in a clinical whole body MR system in vitro. In vivo feasibility to measure quantitative T(2) maps was shown for human patellar cartilage. The relaxation times of cartilage with collagen in the radial zone oriented perpendicular to the magnetic field increased from approximately 10 ms near the bone to approximately 60 ms near the articular surface. Cartilage water content of the tibial plateau and femoral condyles could be determined from the correlation with T(2) (R(2) = 0.71) with an error of approximately 2 wt.%. In vivo, directional variation would need to be considered. If confirmed in vivo, T(2) measurements could potentially serve as a non-invasive tool for the evaluation of the status and distribution of water content in articular cartilage. PMID- 10788721 TI - Quantitative mapping of transverse relaxivity (1/T(2)) in hepatic iron overload: a single spin-echo imaging methodology. AB - Recent research into the non-invasive assessment of hepatic iron concentrations using magnetic resonance imaging has shown that the proton transverse relaxivity (1/T(2)) varies linearly with liver iron concentration. However, the development of an image-based system for the assessment of hepatic iron distribution has been confounded by the presence of motion induced artifacts in the T(2)-weighted images. We report on the development of a single spin-echo imaging methodology that enables the generation of transverse relaxivity maps over the liver. A simple smoothing technique is used to accommodate the image intensity perturbations caused by abdominal motion. The relaxivity maps are consistent with the variation of iron concentration throughout the liver. A Parzen density estimate and histogram of the relaxivity distribution are generated to assist in the visual assessment of the degree and variability of T(2) shortening with liver iron loading. It was found that one or two Gaussian functions could be used to characterize the relaxivity distributions with a small number of parameters. We propose that this methodology may be used in the clinical setting to monitor hepatic iron concentrations in the advent of an accurate transverse relaxivity calibration curve. PMID- 10788722 TI - Automatic measurement of changes in brain volume on consecutive 3D MR images by segmentation propagation. AB - This article presents a technique to automatically measure changes in the volume of a structure of interest in successive 3D magnetic resonance (MR) images and its application in the study of the brain and lateral cerebral ventricles. The only manual step is a segmentation of the structure of interest in the first image. The analysis comprises, first, precise rigid co-registration of the time series of images; second, computation of residual deformations between pairs of images; third, automatic quantification of the volume change, obtained by propagation of the segmentation of the structure of interest through the series of MR images. This approach has been applied to monitor changes in the volume of the brain and lateral cerebral ventricles in a healthy subject and a patient with primary progressive aphasia (PPA). Results are consistent with those obtained by application of the boundary shift integral (BSI) and by stereology in the same subjects. PMID- 10788723 TI - A (1)H magnetic resonance spectroscopy study of aging in parietal white matter: implications for trials in multiple sclerosis. AB - 1H magnetic resonance spectroscopy (MRS) provides a unique tool to detect and quantify brain metabolites. In multiple sclerosis it can be used to investigate axonal loss or dysfunction through measurement of N-acetyl aspartate (NAA), a neuronal marker. Previous studies in adults have reported variable effects of aging on metabolite concentrations but have predominantly focused on changes in the elderly. This study has examined a younger adult age group to provide a reference database more applicable to the multiple sclerosis population. Single voxel (1)H MRS was carried out in 44 subjects between 22 and 62 years of age. Sixteen subjects underwent repeat examination after one year. Absolute concentrations of NA (the sum of NAA and N-acetyl aspartate glutamate), NAA, creatine/phosphocreatine (Cr), choline containing compounds (Cho) and myo inositol (mI) were measured. NA, NAA and mI concentrations did not correlate with age but there were significant correlations between age and Cr (r = 0.43, p = 0.004) and Cho (r = 0.38, p = 0. 011) concentrations. No significant differences in metabolite concentrations were seen over one year. This study provides evidence that age-related changes of metabolite concentrations occur even in a young to middle aged adult population. This emphasizes the need to perform absolute quantification of metabolite concentrations rather than ratios and the importance of age-matching in (1)H MRS studies of multiple sclerosis. PMID- 10788724 TI - Restoration of low resolution metabolic images with a priori anatomic information: 23Na MRI in myocardial infarction. AB - A new iterative extrapolation image reconstruction algorithm is presented, which enhances low resolution metabolic magnetic resonance images (MRI) with information about the bounds of signal sources obtained from a priori anatomic proton ((1)H) MRI. The algorithm ameliorates partial volume and ringing artefacts, leaving unchanged local metabolic heterogeneity that is present in the original dataset but not evident at (1)H MRI. Therefore, it is ideally suited to metabolic studies of ischemia, infarction and other diseases where the extent of the abnormality at (1)H MRI is uncertain. The performance of the algorithm is assessed by simulations, MRI of phantoms, and by surface coil 23Na MRI studies of canine myocardial infarction on a clinical scanner where the injury was not evident at (1)H MRI. The algorithm includes corrections for transverse field inhomogeneity, and for the leakage of intense signals into regions of interest such as 23Na MRI signals from ventricular blood ringing into the myocardium. The simulations showed that the algorithm reduced ringing artefacts by 15%, was stable at low SNR ( approximately 7), but is sensitive to the positioning of the (1)H MRI boundaries. The 23Na MRI showed hyperenhancement of regions identified as infarcted at post-mortem histological staining. The areas of hyperenhancement were measured by five independent observers in four 23Na images of infarction reconstructed with and without the algorithm. The infarct areas were correlated with areas determined by post-mortem histological staining with coefficient 0.85 for the enhanced images, compared to 0.58 with the conventional images. The scatter in the amplitude and in the area measurements of ischemia-associated hyper-enhancement in 23Na MRI was reduced by the algorithm by 1.6-fold and by at least 3-fold, respectively, demonstrating its ability to substantially improve quantification of the extent and intensity of metabolic changes in injured tissue that is not evident by (1)H MRI. PMID- 10788725 TI - Stable periodic vortex shedding studied using computational fluid dynamics, laser sheet flow visualization, and MR imaging. AB - Recirculating and detached flow patterns close to the carotid bifurcation are thought to play an important role in the development of carotid stenoses by promoting atherosclerosis. The aim of this study was to investigate a flow regime with strong transient characteristics, including vortex shedding and transport to develop methodologies appropriate to the analysis of carotid stenoses. The existence of a regular periodic vortex street behind a cylindrical flow obstruction was predicted and analysed in detail by Theodore van Karman in the early 20th century. This model was chosen in our study for both ease of phantom construction and of theoretical modelling using finite element computational fluid dynamics (CFD). The results of the theoretical calculations have been compared with two methods of flow visualization-laser sheet imaging and real-time echo planar magnitude MR imaging. Flow was investigated over a range of Reynold's number from 40 through 400 through which vortex shedding is predicted. Good overall agreement was obtained between the theoretical (16 mm-CFD) and experimental (16+/-2 mm-Laser, 17+/-2 mm-MRI) estimates of the Karman Vortex street wavelength for a Reynolds number of 200. PMID- 10788726 TI - Artifacts caused by transcranial magnetic stimulation coils and EEG electrodes in T(2)*-weighted echo-planar imaging. AB - We investigated the effects of transcranial magnetic stimulation (TMS) coils and electroencephalographic (EEG) electrodes on T(2)*-weighted echo-planar images (EPI) at 2.0 T (gradient-echo EPI, mean TE = 53 ms, 2x2x4 mm(3)). In comparison with anatomic gradient-echo images (3D FLASH, TE = 4 ms, 1x1x1 mm(3)), T(2)* weighted EPI acquisitions of a water-filled spherical phantom revealed severe signal losses and geometric distortions in the vicinity of TMS coils. Even remote effects were observed for image orientations perpendicular to the coil plane. EEG electrodes and the fixation gel caused milder localized distortions. In humans, complications were avoided by the large distance between the TMS coil and the cortical surface and when using an EPI orientation parallel to the plane of the coil. It is concluded that T(2)*-weighted EPI studies of human brain function may be performed without distortions caused by TMS coils and EEG electrodes. PMID- 10788727 TI - Automatic localization of EEG electrode markers within 3D MR data. AB - The electrical activity of the brain can be monitored using ElectroEncephaloGraphy (EEG). From the positions of the EEG electrodes, it is possible to localize focal brain activity. Thereby, the accuracy of the localization strongly depends on the accuracy with which the positions of the electrodes can be determined. In this work, we present an automatic, simple, and accurate scheme that detects EEG electrode markers from 3D MR data of the human head. PMID- 10788728 TI - A comparison of different approaches to promote community-wide dietary change. AB - BACKGROUND: Because public health education funds are limited, it is important to determine which methods are most effective for promoting healthy lifestyles to communities. We conducted interventions in two communities to further examine the effectiveness of various educational approaches for communicating the "1% Or Less" message to switch from high-fat (whole or 2%) to low-fat (1% or fat-free) milk. METHODS: One intervention used public relations and community-based educational activities in supermarkets, schools, worksites, and other community settings. The other used paid advertising in the absence of other programming. We used telephone surveys and supermarket milk sales data, collected before and after each campaign and in a comparison community, to determine changes in milk usage patterns. RESULTS: After the campaign of community-based educational programs and public relations activities, the proportion of high-fat milk drinkers who reported drinking low-fat milk was 19.6% compared with 6.8% for the comparison city (p<0.0001). After the advertising-only campaign, 12.8% of high fat milk drinkers reported drinking low-fat milk (p<0.01). Although supermarkets experienced increases in low-fat milk sales after both campaigns, the results were not statistically significant. CONCLUSIONS: The results show how well designed public relations activities can attract news coverage and provide further evidence that such coverage can be an important component of health promotion campaigns. Although the use of paid advertising in the absence of other media or programming appeared to change milk-drinking habits, the results were not sustained after the ads stopped airing. PMID- 10788729 TI - Do changes in physical activity lead to dietary changes in middle and old age? AB - INTRODUCTION: Research examining the relationship between dietary factors and physical activity patterns has yielded conflicting findings. Few prospective studies have examined whether individuals who change their level of physical activity make corresponding changes in their diet. METHODS: The current study examined, in two randomized clinical trials of physical activity, whether self report of dietary factors changed over a 1-year period, whether these changes differed by gender or exercise-intervention group, and whether changes in physical activity led to changes in dietary factors. Participants included 268 men and women aged 50-65 years who participated in the Stanford-Sunnyvale Health Improvement Project I (SSHIP-I) and 103 men and women aged 65-82 years who participated in SSHIP-II. In both trials we recruited participants through random digit telephone dialing and citywide promotion. RESULTS: Overall, participants adopted healthier diets over the 1-year period. The SSHIP-I participants decreased their consumption of total calories, total fat, saturated fat, protein, and cholesterol. The SSHIP-II participants decreased their consumption of high fat, high-cholesterol foods. The SSHIP-I women were more likely than men to decrease their intake of total fat and protein (p < 0.01), as well as total calories, saturated fat, and cholesterol (p < 0.05). However, we found that changes in physical activity were not associated with changes in self-report measures of dietary habits. CONCLUSIONS: We found no evidence to support the idea that changes in physical activity are a gateway for changes in dietary factors in middle-aged and older adults. PMID- 10788731 TI - Practical nutrition assessment in primary care settings: a review. AB - BACKGROUND: Poor dietary and physical activity habits account for 300,000 deaths per year. Interventions in the primary care setting offer promise for effecting change on a broad scale. However, to conduct counseling, primary care providers need quick, accurate, and efficient assessment tools that are practical for a health care setting. METHODS: We reviewed 18 dietary measures for use in primary care that were brief (less than 50 items) and easy to administer, score, and interpret. The macronutrients assessed by the measure and the psychometric properties, if given, are reported. RESULTS: In general, reliability and validity coefficients were slightly lower than more detailed instruments (i.e., food frequency questionnaires, diet recall); however, they are acceptable within this clinical setting. CONCLUSIONS: Providers can benefit from using these tools to assess current dietary patterns and to guide patient counseling. Future measures should focus on additional nutritional topics (e.g., calcium deficiency) and sensitivity to change over time. PMID- 10788730 TI - A rapid food screener to assess fat and fruit and vegetable intake. AB - BACKGROUND: The U.S. Preventive Services Task Force recommends that Americans lower dietary fat and cholesterol intake and increase fiber and fruit/vegetables to reduce prevalence of heart disease, cancer, stroke, hypertension, obesity, and non-insulin-dependent diabetes mellitus in the United States. To provide preventive services to all, a rapid, inexpensive, and valid method of assessing dietary intake is needed. METHODS: We used a one-page food intake screener based on national nutrition data. Respondents can complete and score the screener in a few minutes and can receive immediate, brief feedback. Two hundred adults self administered the food screener. We compared fat, fiber, and fruit/vegetable intake estimates derived from the screener with estimates from a full-length, 100 item validated questionnaire. RESULTS: The screener was effective in identifying persons with high-fat intake, or low-fruit/vegetable intake. We found correlations of 0.6-0.7 (p<0. 0001) for total fat, saturated fat, cholesterol, and fruit/vegetable intake. The screener could identify persons with high percentages of calories from fat, total fat, saturated fat, or cholesterol, and persons with low intakes of vitamin C, fiber, or potassium. CONCLUSIONS: This screener is a useful tool for quickly monitoring patients' diets. The health care provider can use it as a prelude to brief counseling or as the first stage of triage. Persons who score poorly can be referred for more extensive evaluation by low-cost paper-and-pencil methods. Those who still have poor scores at the second stage ultimately can be referred for in-person counseling. PMID- 10788732 TI - A survey of smoking cessation knowledge, training, and practice among U.S. Army general medical officers. AB - BACKGROUND: Historically, cigarette-smoking rates have been higher among military personnel than among civilians, although recently these rates have decreased. METHODS: In March 1997, a questionnaire assessing (1) training received on smoking cessation, (2) objective knowledge of smoking-cessation techniques, (3) frequency of practice habits, and (4) personal tobacco use among physicians, was successfully mailed to 232 of the total population of 279 Army general medical officers (GMOs). RESULTS: One-hundred-fifty (65%) GMOs returned questionnaires. Of these, 3.3% reported personal cigarette smoking, and 7.3% regularly used smokeless tobacco. During internship, few (13%) GMOs received smoking-cessation training. Primary care programs provided training more frequently than did surgery internship programs. The mean score on the objective knowledge portion was 72%. GMOs had a variable practice pattern in their use of smoking-cessation techniques (percent answering "usually" or "always"): helping patients set quit dates (35%), offering to prescribe the nicotine patch (59%), referring patients to a behavior-modification program (86%). Physicians who received training during internship were significantly more likely (p < 0.01) to help their patients set a quit date. Training did not result in a statistically increased frequency of other practice habits. CONCLUSIONS: GMOs received minimal training on smoking cessation during internship. GMOs refer patients to smoking-cessation classes, reflecting the strategy of the Army Health Promotion program. Strategies to increase the frequency that GMOs prescribe nicotine replacement and assist patients in setting a quit date are needed. Military smoking-cessation efforts may provide valuable lessons for the civilian community. PMID- 10788733 TI - Smoking cessation with and without assistance: a population-based analysis. AB - OBJECTIVE: To examine usage rates of smoking-cessation assistance and to compare the success rate of those who used assistance with the success rate of those who did not. METHODS: The data come from the 1996 California Tobacco Survey, a random sample of 4480 individuals (18 years or older) who tried to quit smoking in the 12 months before the survey. We calculated population estimates for demographics, smoking histories, rate of using assistance, and abstinence rates. RESULTS: One fifth (19.9%) of those who attempted to quit smoking used one or more forms of assistance: self-help, counseling, and/or nicotine replacement therapy (NRT). Heavy smokers were more likely to use assistance than were light smokers. Women were more likely to use assistance than were men, and usage increased with age. Whites were more likely to use NRT than were other ethnic groups. Overall, those who used assistance had a higher success rate than those who did not; the 12 month abstinence rates were 15.2% and 7.0%, respectively. CONCLUSIONS: Use of assistance for smoking cessation has increased over recent years, from 7.9% in 1986 to 19.9% in 1996. The use of assistance is associated with a greater success rate. Anti-tobacco campaigns in California and increased availability of multiple forms of assistance probably facilitated the use of assistance and successful quitting for those using assistance. PMID- 10788734 TI - Longitudinal prediction of sexually transmitted diseases among adolescents: results from a national survey. AB - BACKGROUND: Although adolescent use of condoms has been increasing, incidence of sexually transmitted diseases (STDs) among young people remains high. To identify adolescent behavioral risk factors for acquiring STDs, this study assessed adolescent self-reports of acquired chlamydia, gonorrhea, syphilis, and trichomoniasis within 1 year after a baseline interview. METHODS: We used data from the National Longitudinal Survey of Adolescent Health for this study. Data were collected in the homes of survey respondents, using audio-computer-assisted self-interview (audio-CASI) technology and interviews. Participants were enrolled in grades 7-11 from 134 U.S. schools. A cohort of 4593 sexually experienced adolescents was followed for 1 year. We conducted separate analyses for both genders. RESULTS: About 3.1% of the male adolescents and nearly 4.7% of the female adolescents reported having had at least one STD after the baseline interview. For both genders, self-reported STD infection before baseline interview was the best predictor of self-reported STD infection 1 year after baseline interview. Female adolescents were more likely to report diagnosis with an STD after baseline if they self-identified as a minority race (other than Asian) and perceived that their mother did not disapprove of their having sex. Female adolescents were less likely to report STDs if they perceived that adults care about them. No additional variables predicted STD diagnosis after baseline for male adolescents. CONCLUSIONS: We conclude that past history of STD infection is the most important indicator of subsequent STD infection among adolescents. Thus, this study suggests the benefit of specific clinical efforts designed to promote preventive behavior among adolescents newly diagnosed with an STD. PMID- 10788736 TI - Health literacy and health-related knowledge among persons living with HIV/AIDS. AB - BACKGROUND: Functional health literacy is associated with illness-related knowledge, understanding, and treatment perceptions for several chronic illnesses. This study examined health literacy in relation to knowledge and understanding of HIV/AIDS. METHODS: Persons living with HIV/AIDS recruited from AIDS service organizations and HIV clinics completed the Test of Functional Health Literacy for Adults (TOFHLA) reading comprehension scale and measures of health status, knowledge and understanding of health status, perceptions of primary care givers, and perceptions of anti-HIV treatments. RESULTS: Eighteen percent of the sample scored below the cutoff for marginal functional health literacy on the TOFHLA. Controlling for years of education, persons of lower health literacy were significantly less likely to have an undetectable HIV viral load, somewhat less likely to know their CD4 cell count and viral load, and lower health-literacy persons who knew their CD4 count and viral load were less likely to understand their meaning. Lower health literacy was also related to misperceptions that anti-HIV treatments reduce risks for sexually transmitting HIV and beliefs that anti-HIV treatments can relax safer-sex practices. CONCLUSIONS: Poor health literacy creates barriers to fully understanding one's health, illness, and treatments. Misperceptions of treatment in the case of HIV infection creates danger for potentially transmitting treatment-resistant strains of HIV. These results have implications for patient education and treatment programming for people who have poor health-literacy skills and are living with HIV/AIDS. PMID- 10788735 TI - Decline in physician referrals to health department clinics for immunizations: the role of vaccine financing. AB - BACKGROUND: Physicians frequently refer children to health department clinics (HDCs) for immunizations because of high out-of-pocket costs to parents and poor reimbursement for providers. Referrals for immunizations can lead to scattered care. In 1994, two vaccine financing reforms began in New York State that reduced patient costs and improved provider reimbursement: the Vaccines for Children Program (VFC, mostly for those on Medicaid and uninsured) and a law requiring indemnity insurers to cover childhood immunizations and preventive services. OBJECTIVE: To measure reported changes in physician referrals to HDCs for immunizations before and after the vaccine financing reforms. DESIGN: In 1993, a self-administered survey measured immunization referral practices of primary care physicians. In 1997, we resurveyed respondents of the 1993 survey to evaluate changes in referrals. SETTING/ PARTICIPANTS: Three hundred twenty-eight eligible New York State primary care physicians (65% pediatricians and 35% family physicians) who responded to the 1997 follow-up immunization survey (response rate of 82%). RESULTS: The proportion of physicians reporting that they referred some or all children out for immunizations decreased from 51% in 1993 to 18% in 1997 (p<0.001). In 1997, physicians were more likely to refer if they were family physicians (28% vs. 13%,p<0.01), or did not obtain VFC vaccines (29% vs. 13%,p<0.001). According to physicians who referred in 1993, decreased referrals in 1997 were due to the new insurance laws (noted by 61%), VFC (60%), Child Health Plus (a statewide insurance program for poor children, 28%), growth in commercial managed care (23%), Medicaid managed care (19%), and higher Medicaid reimbursement for immunizations that is due to VFC (18%). For physicians noting a decline in referrals, the magnitude of the decline was substantial-60% fewer referrals for VFC-eligible patients and 50% fewer for patients eligible under the new insurance law. CONCLUSIONS: Vaccine financing reforms decreased the proportion of physicians who referred children to HDCs for immunizations, and may have reduced scattering of pediatric care. PMID- 10788737 TI - Effects of a short-term health promotion intervention for a predominantly African American group of stroke survivors. AB - BACKGROUND: The study examined the effects of a 12-week health promotion intervention for a predominantly urban African-American population of stroke survivors. DESIGN: A pre-test/post-test lag control group design was employed. PARTICIPANTS/SETTING: Participants were 35 stroke survivors (9 male, 26 female) recruited from local area hospitals and clinics. MAIN OUTCOME MEASURES: Biomedical, fitness, nutritional, and psychosocial measures were employed to assess program outcomes. RESULTS: Treatment group made significant gains over lag controls in the following areas: (1) reduced total cholesterol, (2) reduced weight, (3) increased cardiovascular fitness, (4) increased strength, (5) increased flexibility, (6) increased life satisfaction and ability to manage self care needs, and (7) decreased social isolation. CONCLUSION: A short-term health promotion intervention for predominantly African-American stroke survivors was effective in improving several physiological and psychological health outcomes. PMID- 10788739 TI - The effect of audit and feedback on immunization delivery: a systematic review. AB - OBJECTIVE: To assess the effective of audit and feedback (A&F) on immunization delivery by health care professionals. DESIGN: Systematic review of published literature. MAIN OUTCOME MEASURES: Changes in immunization rates. METHODS: We searched Medline between 1966 and 1997. We obtained additional studies from back searching reference lists and the files of study collaborators. We included studies that were written in English, that included audit and feedback in at least one arm of the study, that studied universally recommended childhood or adult vaccines, and that provided immunization coverage data. Two reviewers read studies independently and abstracted using a validated checklist. Study quality was assessed using criteria standardized by the Cochrane Collaboration. Differences between reviewers were resolved by consensus. RESULTS: The search process resulted in 60 citations; 44 were fully reviewed and 15 met eligibility criteria. Five were randomized trials. Twelve of the fifteen studies found that A&F, alone or in combination with other interventions, were associated with improvements in immunization rates. The magnitude of the effect varied from -17% to +49% change. Study design heterogeneity precluded statistical pooling of study results. CONCLUSIONS: The evidence available from published studies suggests that A&F alone may be an effective strategy for improving immunization rates. The number of well-conducted studies is small, and the effect is variable. Additional well-designed studies are needed to identify the independent effects of A&F, optimal format and frequency of A&F, and to examine its long-term effect on provider immunization practices and costs. PMID- 10788740 TI - Nutrition in the prevention of disease: current issues and concepts. PMID- 10788738 TI - Massachusetts' advertising against light cigarettes appears to change beliefs and behavior. AB - OBJECTIVES: This study examined the effects of advertising directed against light cigarettes (lights). METHODS: In a quasi-experimental post-test-only design, smokers and ex-smokers (6 glycosylation, a cis-Golgi-specific modification, but not alpha1-->;3 glycosylation, a medial Golgi-specific modification. In contrast, a single-copy expression of this chimera does not lead to the cis-Golgi-specific modification. These findings suggest that, when expressed on a multicopy plasmid, a fraction of this chimera is transported from the ER to the cis-Golgi compartment and subsequently recycled to the ER, but when expressed on a single-copy plasmid, no significant transport of this protein from the ER takes place. We thus suggest the possibility that cytochrome P450 is retained in the ER by a saturable static mechanism. PMID- 10788784 TI - New world monkey pepsinogens A and C, and prochymosins. Purification, characterization of enzymatic properties, cDNA cloning, and molecular evolution. AB - Pepsinogens A and C, and prochymosin were purified from four species of adult New World monkeys, namely, common marmoset (Callithrix jacchus), cotton-top tamarin (Saguinus oedipus), squirrel monkey (Saimiri sciureus), and capuchin monkey (Cebus apella). The occurrence of prochymosin was quite unique since this zymogen is known to be neonate-specific and, in primates, it has been thought that the prochymosin gene is not functional. No multiple form has been detected for any type of pepsinogen except that two pepsinogen-A isozymogens were identified in capuchin monkey. Pepsins A and C, and chymosin hydrolyzed hemoglobin optimally at pH 2-2.5 with maximal activities of about 20, 30, and 15 units/mg protein. Pepsins A were inhibited in the presence of an equimolar amount of pepstatin, and chymosins and pepsins C needed 5- and 100-fold molar excesses of pepstatin for complete inhibition, respectively. Hydrolysis of insulin B chain occurred first at the Leu15-Tyr16 bond in the case of pepsins A and chymosins, and at either the Leu15-Tyr16 or Tyr16-Leu17 bond in the case of pepsins C. The presence of different types of pepsins might be advantageous to New World monkeys for the efficient digestion of a variety of foods. Molecular cloning of cDNAs for three types of pepsinogens from common marmoset was achieved. A phylogenetic tree of pepsinogens based on the nucleotide sequence showed that common marmoset diverged from the ancestral primate about 40 million years ago. PMID- 10788785 TI - Sequence motif-specific assignment of two [2Fe-2S] clusters in rat xanthine oxidoreductase studied by site-directed mutagenesis. AB - The sequence motif-specific assignment of the two distinct [2Fe-2S] clusters in rat xanthine oxidoreductase (XOR) was unequivocally established by site-directed mutagenesis of recombinant enzymes expressed in a baculovirus-insect cell system and electron paramagnetic resonance (EPR) spectroscopy. The conserved cysteine residues, including Cys-115, in the unusual C-terminal -Cys-Xaa(2)-Cys-//-Cys Xaa(1)-Cys- motif serve as ligands to the Fe/S I center, which is probably located in close proximity to the Mo-pterin center. Other conserved cysteine residues, including Cys-43 and Cys-51, in the N-terminal plant ferredoxin-like motif serve as ligands to the Fe/S II center, which is distantly located from the Mo-pterin center. The present sequence motif-specific assignment of the Fe/S I and II centers is discussed in the light of the structural features of XOR. PMID- 10788786 TI - Polyketide synthase gene coupled to the peptide synthetase module involved in the biosynthesis of the cyclic heptapeptide microcystin. AB - The peptide synthetase gene operon, which consists of mcyA, mcyB, and mcyC, for the activation and incorporation of the five amino acid constituents of microcystin has been identified [T. Nishizawa et al. (1999) J. Biochem. 126, 520 529]. By sequencing an additional 34 kb of DNA from microcystin-producing Microcystis aeruginosa K-139, we identified the residual microcystin synthetase gene operon, which consists of mcyD, mcyE, mcyF, and mcyG, in the opposite orientation to the mcyABC operon. McyD consisted of two polyketide synthase modules, and McyE contained a polyketide synthase module at the N-terminus and a peptide synthetase module at the C-terminus. McyF was found to exhibit similarity to amino acid racemase. McyG consisted of a peptide synthetase module at the N terminus and a polyketide synthase at the C-terminus. The microcystin synthetase gene cluster was conserved in another microcystin-producing strain, Microcystis sp. S-70, which produces Microcystin-LR, -RR, and -YR. Insertional mutagenesis of mcyA, mcyD, or mcyE in Microcystis sp. S-70 abolished microcystin production. In conclusion, the mcyDEFG operon is presumed to be responsible for 3-amino-9 methoxy-2,6, 8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda) biosynthesis, and the incorporation of Adda and glutamic acid into the microcystin molecule. PMID- 10788787 TI - Distinctive functions of Syk N-terminal and C-terminal SH2 domains in the signaling cascade elicited by oxidative stress in B cells. AB - Syk plays a crucial role in the transduction of oxidative stress signaling. In this paper, we investigated the roles of Src homology 2 (SH2) domains of Syk in oxidative stress signaling, using Syk-negative DT40 cells expressing the N- or C terminal SH2 domain mutant [mSH2(N) or mSH2(C)] of Syk. Tyrosine phosphorylation of Syk in cells expressing mSH2(N) Syk after H(2)O(2) treatment was higher than that in cells expressing wild-type Syk or mSH2(C) Syk. The tyrosine phosphorylation of wild-type Syk and mSH2(C) Syk, but not that of mSH2(N), was sensitive to PP2, a specific inhibitor of Src-family protein-tyrosine kinase. In oxidative stress, the C-terminal SH2 domain of Syk was demonstrated to be required for induction of tyrosine phosphorylation of cellular proteins, phospholipase C (PLC)-gamma2 phosphorylation, inositol 1,4, 5-triphosphate (IP(3)) generation, Ca(2)(+) release from intracellular stores, and c-Jun N terminal kinase activation. In contrast, in mSH2(N) Syk-expressing cells, tyrosine phosphorylation of intracellular proteins including PLC-gamma2 was markedly induced in oxidative stress. The enhanced phosphorylation of mSH2(N) Syk and PLC-gamma2, however, did not link to Ca(2)(+) mobilization from intracellular pools and IP(3) generation. Thus, the N- and C-terminal SH2 domains of Syk possess distinctive functions in oxidative stress signaling. PMID- 10788788 TI - Enhanced ability of heparin-carrying polystyrene (HCPS) to bind to heparin binding growth factors and to inhibit growth factor-induced endothelial cell growth. AB - Heparin-carrying polystyrene (HCPS) consists of low-molecular-weight heparin chains enriched in trisulfated disaccharide structures linked to a polystyrene core. In this study, the interactions between HCPSs of various molecular weights and heparin-binding growth factors, VEGF(165), FGF-2, and HGF, were compared to the interactions of the same factors with native heparin, periodate-oxidized heparin (IO(4)-heparin) and periodate-oxidized alkaline-degraded heparin (IO(4) LMW-heparin). The binding of each growth factor to heparin-agarose beads (heparin beads) was more strongly inhibited by HCPSs in a molecular weight-dependent manner than by native heparin or the modified heparins, indicating a stronger interaction between HCPS and these growth factors. HCPSs also inhibit heparin binding growth factor-induced endothelial cell growth in a molecular weight dependent manner much more strongly than the native or modified heparins. However, HCPSs did not inhibit the mitogenic activity of VEGF(121), which has a non-heparin-binding nature. Thus, HCPSs exhibit enhanced abilities to interact with each of the heparin-binding growth factors studied and to inhibit heparin binding growth factor-induced endothelial cell proliferation in a molecular weight-dependent manner. These effects might be ascribed to the heparin clustering effect of HCPSs. PMID- 10788789 TI - Structure of the pig sterol 14alpha-demethylase (CYP51) gene and its expression in the testis and other tissues. AB - A cDNA coding sterol 14alpha-demethylase (CYP51), which was isolated from a pig liver cDNA, contained a 1,512 bp open reading frame and a 758 bp 3'-untranslated region. The deduced amino acid sequence was 94% identical to those of human and rat CYP51s. The pig CYP51 gene spanned about 21 kb and was divided into 10 exons. The sites of exon-intron junctions were completely identical to those in the human and rat CYP51 genes. Five GC boxes, but not a TATA box, were found in the 5'-flanking region of the gene, and cyclic AMP and sterol responsive elements were also found in this region. The main transcription start site determined with the 5'-RACE method with poly(A)(+) RNA from the liver and testis was located at 143 nucleotides upstream from the initiation codon in both tissues. Northern blot analysis revealed that an approximately 2.4 kb mRNA, which is produced through the use of a polyadenylation signal (AATAAA) located at 740 nucleotides downstream of the stop codon, was expressed in all the tissues examined in pigs: The mRNA levels were much higher in the liver and testis than in the kidney, lung, and epididymis. Furthermore, after the onset of spermatogenesis, a smaller size of mRNA (about 1.8 kb) was found in the testis but not in the epididymis. The 1.8 kb mRNA was produced through the use of an unusual polyadenylation signal (AAGAAA) located at 28 nucleotides downstream of the stop codon. PMID- 10788790 TI - Rescuing activity of galactoglycerolipids from cellular lesions induced by 5 aminolevulinic acid. AB - An anti-oxygen radical reagent of a bacterial metabolite, M874 monogalactoglycerolipid (di-O-12-methyl-tetradecanoyl-3-O-beta-D-galactopyranosyl sn-glycerol ), was tested for its ability to protect two organisms against cellular lesions induced by 5-aminolevulinic acid (ALA) and light. In Corynebacterium flavescens ATCC 10340, extracellular uroporphyrin and coproporphyrin were the main porphyrin products. Although less than 2 mM ALA increased porphyrin synthesis, ALA levels above 3 mM inhibited the synthesis. Depending on the light intensity, the amount of porphyrin decreased and ALA induced cytotoxicity increased. The lesion was more severe in the case of coproporphyrin than uroporphyrin. The porphyrin lesion produced in low intensity light (300 lx) was considerably reduced by 100 microM M874 glycolipid, although the reduction in intense light (3,000 lx) was restricted to a lower level. Similar results were obtained with radish (Raphanus sativus). The ALA concentration that inhibited porphyrin synthesis and stem growth was similar to that seen with C. flavescens. Although the exogenous addition of M874 glycolipid to the radish did not prevent ALA-induced cellular injury, the co-culture of radish and a glycolipid producing bacterium (Microbacterium sp. M874) resulted in a significant prevention of cellular injury. This was true only under enforced adhesion conditions through the action of a polysaccharide flocculant H12. Some species of monogalactoglycerolipids were found in Corynebacterium and radish that showed prominent oxygen radical-protecting activities similar to that of M874 glycolipid. These monogalactoglycerolipids might function in vivo as agents to prevent ALA-induced cytological lesions, although the concentrations were low in Corynebacterium and radish. PMID- 10788791 TI - Membrane enzyme systems responsible for the Ca(2+)-dependent phosphorylation of Ser(27), the independent phosphorylation of Tyr(10) and Tyr(7), and the dephosphorylation of these phosphorylated residues in the alpha-chain of H/K ATPase. AB - H/K-ATPase preparations (the G1 membrane) from pig stomach contain both kinases and phosphatases and show reversible phosphorylation of Tyr(7), Tyr(10), and Ser(27) residues of the alpha-chain of H/K-ATPase. The Tyr-kinase is sensitive to genistein and quercetin and recognized by anti-c-Src antibody. The Ser-kinase is dependent on Ca(2)(+) (K(0.5) = 0.9 microM), sensitive to a PKC inhibitor, and recognized by antibodies against PKCalpha and PKCbetaII. The addition of 3-[(3 cholamidopropyl)dimethylammonio]-1-propane-sulfonic acid (CHAPS) caused a dramatic increase in the phosphorylation of added synthetic copolymer substrates and permitted the phosphorylation of maltose-binding proteins fused with the N terminal domain of alpha-chains. The phosphotyrosine phosphatase was inhibited by vanadate. The phosphoserine phosphatase was inhibited by okadaic acid and by inhibitor-2. The presence of protein phosphatase-1 was immunologically detected. Column chromatographic separation of CHAPS-solubilized G1 membrane and others indicate the apparent molecular weight of the Src-kinase to be approximately 60 kDa, the PKCalpha and/or PKCbII to be approximately 80 kDa, the Tyr-phosphatase to be 200 kDa, and PP-1 to be approximately 35 kDa. These data show that these membrane-bound enzyme systems are in sufficiently close proximity to be responsible for reversible phosphorylation of Tyr(7), Tyr(10), and Ser(27) of the catalytic subunit of membrane H/K-ATPase in parietal cells, the physiological role of which is unknown. PMID- 10788792 TI - Divalent forms of CC49 single-chain antibody constructs in Pichia pastoris: expression, purification, and characterization. AB - Single-chain variable fragments (scFvs) are tumor-recognition units that hold enormous potential in antibody-based therapeutics. Their clinical applications, however, require the large scale production and purification of biologically active recombinant scFvs. In the present study, we engineered and expressed divalent non-covalent [(scFv)(2)-His(6)] and covalent [sc(Fv)(2)-His(6)] scFvs of a tumor-associated monoclonal antibody (MAb) CC49 in Pichia pastoris. The purity and immunoreactivity of the scFvs were analyzed by SDS-PAGE, HPLC, and competitive ELISA. The binding affinity constant (K(A)), determined by surface plasmon resonance analysis using BIAcore, was 4.28 x 10(7), 2.75 x 10(7), and 1.14 x 10(8) M(-1) for (scFv)(2)-His(6), sc(Fv)(2)-His(6), and CC49 IgG, respectively. The expression of scFvs in P. pastoris was 30 to 40-fold higher than in Escherichia coli. Biodistribution studies in athymic mice bearing LS-174T human colon carcinoma xenografts showed equivalent tumor-targeting of CC49 dimers generated in yeast (scFv)(2)-His(6) and bacteria (scFv)(2) with 12.52% injected dose/gram (%ID/g) and 11. 42%ID/g, respectively, at 6 h post-injection. Interestingly, the pharmacokinetic pattern of dimeric scFvs in xenografted mice exhibited a slower clearance of His-tagged scFvs from the blood pool than scFvs lacking the His-tag (0.1 >/= p >/= 0.05). In conclusion, improved yields of divalent scFvs were achieved using the P. pastoris expression/secretion system. The in vitro and in vivo properties of these scFvs suggest possible therapeutic applications. PMID- 10788793 TI - Regulatory role of C-terminal residues of SulA in its degradation by Lon protease in Escherichia coli. AB - The SulA protein is a cell division inhibitor in Escherichia coli, and is specifically degraded by Lon protease. To study the recognition site of SulA for Lon, we prepared a mutant SulA protein lacking the C-terminal 8 amino acid residues (SA8). This deletion protein was accumulated and stabilized more than native SulA in lon(+) cells in vivo. Moreover, the deletion SulA fused to maltose binding protein was not degraded by Lon protease, and did not stimulate the ATPase or peptidase activity of Lon in vitro, probably due to the much reduced interaction with Lon. A BIAcore study showed that SA8 directly interacts with Lon. These results suggest that SA8 of SulA was recognized by Lon protease. The SA8 peptide, KIHSNLYH, specifically inhibited the degradation of native SulA by Lon protease in vitro, but not that of casein. A mutant SA8, KAHSNLYH, KIASNLYH, or KIHSNAYH, also inhibited the degradation of SulA, while such peptides as KIHSNLYA did not. These results show that SulA has the specified rows of C terminal 8 residues recognized by Lon, leading to facilitated binding and subsequent cleavage by Lon protease both in vivo and in vitro. PMID- 10788794 TI - Molecular cloning and genomic analysis of mouse GalNAc alpha2, 6 sialyltransferase (ST6GalNAc I). AB - cDNA clones encoding mouse GalNAc alpha2,6-sialyltransferase (ST6GalNAc I) were isolated from a mouse submaxillary gland cDNA library. The deduced amino acid sequence of cDNA clones is 526 amino acids in length and has highly conserved motifs among sialyl transferases, sialyl motifs L, S, and VS. The expressed recombinant enzyme exhibited similar substrate specificity to chicken ST6GalNAc I. The mouse ST6GalNAc I gene was expressed in submaxillary gland, mammary gland, colon, and spleen. The mouse ST6GalNAc I gene was also cloned from a mouse genomic library, which was divided into 9 exons spanning over 8 kilobases of genomic DNA. The genomic structure of the mouse ST6GalNAc I gene was similar to that of the mouse ST6GalNAc II gene. Unlike the ST6GalNAc II gene, however, which has a housekeeping gene-like promoter with GC-rich sequences, the ST6GalNAc I gene has two promoters and they do not contain GC-rich sequences but contain putative binding sites for tumor-associated transcription factors such as c-Myb, c-Myc/Max, and c-Ets. Analysis of the 5'-RACE PCR products suggested that the mouse ST6GalNAc I gene expression is regulated by these two promoters in tissue specific manners. PMID- 10788795 TI - Characterization of three putative sub-domains in the signal-input domain of the ArcB hybrid sensor in Escherichia coli(1). AB - The ArcB sensor plays a crucial role in the histidine to aspartate (His-to-Asp) phosphorelay signal transduction, which is involved in the transcriptional regulatory network that allows Escherichia coli cells to sense various respiratory growth conditions. ArcB is one of the best-studied hybrid His-kinases involved in the multi-step His-to-Asp phosphorelay. However, a major question that remains to be elucidated is: how does ArcB sense an anoxic signal? The N terminal region of ArcB is considered to be a signal-input domain, which probably plays a role in such signal-perception. In this study, this N-terminal region of ArcB was dissected into three putative sub-domains, a "transmembrane domain," a "leucine-zipper-like domain, " and a "PAS-like domain." The importance of these structural domains was assessed in vivo and in vitro by systematically analyzing a number of arcB mutants, each of which encodes a mutant ArcB protein having an amino acid substitution or a deletion within one of these sub-domains. The results are discussed with special reference to the nature of the ArcB anaerobic sensor. PMID- 10788796 TI - Irreversible conformational change of bacterio-opsin induced by binding of retinal during its reconstitution to bacteriorhodopsin, as studied by (13)C NMR. AB - We compared (13)C NMR spectra of [3-(13)C]Ala- and [1-(13)C]Val-labeled bacterio opsin (bO), produced either by bleaching bR with hydroxylamine or from a retinal deficient strain, with those of bacteriorhodopsin (bR), in order to gain insight into the conformational changes of the protein backbone that lead to correct folding after retinal is added to bO. The observed (13)C NMR spectrum of bO produced by bleaching is not greatly different from that of bR, except for the presence of suppressed or decreased peak-intensities. From careful evaluation of the intensity differences between cross polarization magic angle spinning (CP MAS) and dipolar decoupled-magic angle spinning (DD-MAS) spectra, it appears that the reduced peak-intensities arise from reduced efficiency of cross polarization or interference of internal motions with proton decoupling frequencies. In particular, the E-F and F-G loops and some transmembrane helices of the bleached bO have acquired internal motions whose frequencies interfere with proton decoupling frequencies. In contrast, the protein backbone of the bO from the retinal-negative cells is incompletely folded. Although it contains mainly a helices, its very broad (13)C NMR signals indicate that its tertiary structure is different from bR. Importantly, this changed structure is identical in form to that of bleached bO from wild-type bR after it was regenerated with retinal in vitro, and bleached with hydroxylamine. We conclude that the binding of retinal is essential for the correct folding of bR after it is inserted in vitro into the lipid bilayer, and the final folded state does not revert to the partially folded form upon removal of the retinal. PMID- 10788797 TI - Activation of phospholipase A(2) by long chain fatty acyl groups involves a novel unstable linkage. AB - The acidic isoform of phospholipase A(2) from Naja mossambica mossambica was activated by treatment with a molar equivalent of oleoyl imidazolide. Modification of the protein was accompanied by 50% quenching of tryptophan fluorescence and a significant red shift. The (3)H(9,10) labeled oleoyl residue was co-eluted with the enzyme during gel filtration in the presence of 20% 1 propanol or excess albumin, both of which remove free oleic acid from the enzyme. In contrast, the adduct was labile as to electrophoresis on SDS-PAGE and acid or alkali urea PAGE. The formation of a covalently linked adduct was demonstrated by electrospray mass spectrometry in the presence of 2% formic acid. No such adduct was formed by the phospholipase A(2) isoform from Naja naja atra, which differs in sequence from the N. mossambica mossambica isoform by seven residues including 2 histidine residues and 1 lysine residue. We conclude that oleoyl imidazolide activates the N. mossambica mossambica enzyme by forming an acyl adduct which is unstable as to protein denaturation. The magnitude of tryptophan fluorescence quenching indicates that the site of acylation lies in the sequence WWHF. PMID- 10788798 TI - Effect of mutagenic replacement of the carboxyl terminal amino acid, val124, on the properties and regeneration of bovine pancreatic ribonuclease A. AB - An important role of C-terminal amino acid residues of bovine pancreatic ribonuclease A (RNase A) in the formation of the three-dimensional structure was previously implied. In this study, we replaced the C-terminal amino acid, Val124, with amino acid residues with different properties by site-directed mutagenesis. The recombinant mutant enzymes were purified and subjected to a refolding study after being converted to a fully reduced and denatured state. There was a significant difference among the mutant enzymes in the rate of recovery of the activity when air oxidation was performed: the rate decreased in the order of V124E, V124L, V124G, V124K, V124A, and V124W. On the other hand, the recovery rates for all the mutant RNase A in the presence of GSH and GSSG were almost the same. The recovered activity of V124E after 24 h incubation reached approximately 90% of that of the wild type enzyme, followed by V124L 80%, V124A and V124W 65%, and V124K and V124G 50%. The duration of the initial lag phase became shorter in the order of V124W, V124A, V124K or V124G, V124E, or V124L. The results imply that the C-terminal amino acid significantly influences the formation of correct disulfide bonds during the refolding process and that the hydrophobic interaction of Val124 is important for efficient packing of the RNase A molecule. PMID- 10788799 TI - IL-1 induces expression of p21(WAF1) independently of p53 in high-passage human embryonic fibroblasts WI38. AB - We tested the effect of IL-1 on the expression of p21(WAF1) in human embryonic fibroblasts WI38. Exposure to IL-1 caused induction of p21(WAF1) protein in high passage WI38 cells but not in early-passage cells. However, IL-1 did not stimulate the transcription of a CAT-reporter gene having two copies of the p53 responsive element on its promoter or the p53-binding capacity of nuclear extracts, although it increased transcriptional rate of p21(WAF1) in these high passage cells. These results suggest that the induction of p21(WAF1) by IL-1 occurs at the transcriptional level, but p53 function is not required in these cells. Further studies found that IL-1 did not cause cell-cycle arrest, and the overexpression of p21(WAF1) resulted in only a slight delay of cell growth, while the level of p21(WAF1) coprecipitated with cyclin-dependent kinase-2 (Cdk2) was increased by IL-1. Moreover, a kinase assay of Cdk2 immunoprecipitates showed that IL-1 did not reduce the kinase activity, and IL-1 did not affect the status of phosphorylation of the retinoblastoma gene product (Rb). These findings imply that despite the induction of p21(WAF1), this cannot fully account for the growth arrest in high-passage WI38 cells. Thus, IL-1 mediates p21(WAF1) induction through a p53-independent pathway(s) in high-passage WI38 cells, but the cell cycle is regulated independently of p21(WAF1). PMID- 10788800 TI - Characterization of ribonuclease HII from Escherichia coli overproduced in a soluble form. AB - Escherichia coli RNase HII is composed of 198 amino acid residues. The enzyme has been overproduced in an insoluble form, purified in a urea-denatured form, and refolded with poor yield [M. Itaya (1990) Proc. Natl. Acad. Sci. USA 87, 8587 8591]. To facilitate the preparation of the enzyme in an amount sufficient for physicochemical studies, we constructed an overproducing strain in which E. coli RNase HII is produced in a soluble form. The enzyme was purified from this strain and its biochemical and physicochemical properties were characterized. The good agreement in the molecular weights estimated from SDS-PAGE (23,000) and gel filtration (22,000) suggests that the enzyme acts as a monomer. From the far-UV circular dichroism spectrum, its helical content was calculated to be 23%. The enzyme showed Mn(2+)-dependent RNase H activity. Its specific activity determined using (3)H-labeled M13 RNA/DNA hybrid as a substrate was comparable to but slightly higher than that of the refolded enzyme, indicating that the enzyme overproduced and purified in a soluble form is more suitable for structural and functional analyses than the refolded enzyme. PMID- 10788801 TI - Identification and characterization of an anterior fat body protein in an insect. AB - We purified a novel protein with a molecular mass of 34 kDa from the Sarcophaga larval fat body. This protein, named AFP (anterior fat body protein), was restricted almost exclusively to the anterior fat body. The AFP content decreased after pupation on disintegration of the fat body tissue. cDNA analysis revealed that this protein consists of 306 amino acid residues and exhibits significant structural similarity with mammalian regucalcin (senescence marker protein-30), a calcium-binding liver protein. However, AFP did not seem to exhibit strong affinity with calcium. These results suggested that a seemingly uniform fat body tissue exhibits a regional difference in its function along the anterior posterior axis. PMID- 10788802 TI - Rho-mediated phosphorylation of focal adhesion kinase and myosin light chain in human endothelial cells stimulated with sphingosine 1-phosphate, a bioactive lysophospholipid released from activated platelets. AB - Since sphingosine 1-phosphate (Sph-1-P) is stored in abundant amounts in blood platelets and released extracellularly upon stimulation, it is important to clarify the effects of this bioactive lysophospholipid on vascular endothelial cells from the viewpoint of platelet-endothelial cell interactions. In this study, we investigated the effects of Sph-1-P on the cytoskeletal remodeling of human umbilical vein endothelial cells (HUVECs). Of a focal adhesion kinase (FAK) family of non-receptor protein-tyrosine kinases, HUVECs were found to express FAK, but scarcely proline-rich tyrosine kinase 2. Sph-1-P induced FAK tyrosine phosphorylation, myosin light chain phosphorylation, and the formation of stress fibers in HUVECs. The specific Rho inactivator C3 transferase from Clostridium botulinum abolished all of these cytoskeletal responses induced by Sph-1-P, while pertussis toxin only partly inhibited FAK tyrosine phosphorylation, and hardly affected myosin light chain phosphorylation and stress fiber formation. In contrast, Sph-1-P-induced intracellular Ca(2)(+) mobilization was suppressed by pertussis toxin, but not at all by C3 exoenzyme. Our results suggest that Sph-1 P, a bioactive lipid released from activated platelets, induces endothelial cell cytoskeletal reorganization, mainly through Rho-mediated signaling pathways. PMID- 10788803 TI - Random expression of main and vomeronasal olfactory receptor genes in immature and mature olfactory epithelia of Fugu rubripes. AB - Main olfactory receptor genes were isolated from a seawater fish, Fugu rubripes (pufferfish), and characterized. Two subfamilies of genes encoding seven transmembrane receptors were identified; one consists of five or more members, termed FOR1-1 to 5 of FOR1 subfamily, and the other appears to be a single copy gene, termed the FOR2 subfamily. FOR1 members show extremely high amino acid sequence similarities of about 95% to one another, and are distantly related to catfish-1 with the highest similarity of 37%. FOR2 shows 43% similarity to goldfish-A28. Phylogenically, both FOR members are categorized among pedigrees of the fish main olfactory receptor family outside the mammalian receptor family, although similarities between Fugu receptors and those of fresh-water fishes are lower than those among fresh-water fishes. In situ hybridization shows that both subfamilies of receptor genes are expressed randomly over the olfactory epithelium throughout all developmental stages, and no segregation of the signals was found. On the other hand, when three members of a vomeronasal olfactory receptor gene family, related to the Ca(2+)-sensing receptor, were used as probes, they were also randomly expressed over the same epithelium as the main olfactory receptors. This is in contrast to the expression profiles observed for zebrafish and goldfish, where the main or vomeronasal olfactory receptors are expressed in segregated patterns. It is thus suggested that the expression pattern of fish olfactory receptors varies depending on the species, although fish olfactory receptors are highly related to one another in their primary structures, and are phylogenically distinct from those of mammals. PMID- 10788804 TI - NTAKalpha and beta isoforms stimulate breast tumor cell growth by means of different receptor combinations. AB - Neural- and thymus-derived activator for ErbB kinases (NTAK) is a recently described member of the neuregulin family that binds directly to ErbB3 and ErbB4 and transactivates ErbB2. Rat NTAK has at least five alternative-spliced isoforms: alpha1, alpha2a, alpha2b, beta, and gamma. In order to understand their biological properties, this study focused on the NTAK alpha2a and beta isoforms, which have different EGF-like domains. The effect of these isoforms on cell growth and tyrosine phosphorylation in human breast cancer cells, MDA-MB-453 and T47D, was examined using the recombinant proteins. In terms of cell growth, NTAKalpha2a and NTAKbeta preferentially stimulate T47D cells and MDA-MB-453 cells, respectively, in a dose-dependent manner. Although both NTAKs induce the highest level of tyrosine phosphorylation of ErbB2, NTAKalpha2a and NTAKbeta preferentially induce ErbB3 and ErbB4 phosphorylation, respectively. Thus, NTAKalpha2a and NTAKbeta stimulate cell growth in different ways, by means of different combinations of receptors. PMID- 10788805 TI - FtsH recognizes proteins with unfolded structure and hydrolyzes the carboxyl side of hydrophobic residues. AB - FtsH of Escherichia coli is an essential membrane-integrated ATP-dependent protease. We cloned a gene for an FtsH homolog (T. FtsH) from Thermus thermophilus HB8, expressed it in E. coli, and purified the expressed protein. ATPase activity of T.FtsH was activated by proteins with unfolded structure ( alpha-casein and pepsin), and T.FtsH digested these proteins in an ATP-, Zn(2+) dependent manner. alpha-Lactalbumin was digested by T.FtsH when it was largely unfolded, but not in its native form. Analysis of the proteolytic products revealed that, in most cases, T.FtsH cleaved the C-terminal side of hydrophobic residues and produced a characteristic set of small peptides (<30 kDa) without releasing a large intermediate. Thus, T.FtsH recognizes the unfolded structure of the proteins and progressively digests them at the expense of ATP. A soluble domain of T.FtsH, which lacked the N-terminal two transmembrane helices, was also prepared but was found to retain neither ATPase nor protease activities. Thus, the membrane segment appeared to be indispensable for these activities of T.FtsH. PMID- 10788806 TI - Five- to fifteen-year follow-up of fresh autologous pericardial valved conduits. AB - OBJECTIVE: Evaluate long-term results of autologous pericardial valved conduits in the pulmonary outflow. METHODS: Between June 1983 and October 1993, 82 conduits were placed in the outflow of the venous ventricle. Patients who received homografts (n = 2 patients), heterografts (n = 3 patients), and valveless conduits (n = 19 patients) and those patients who died within 90 days after the operation were excluded. Fifty-four survivors of pulmonary outflow reconstruction with fresh autologous pericardial valved conduits were followed up from 5 to 15 years (mean, 7.47 +/- 2.8 years). Diagnosis include d -transposition of great arteries (n = 16 patients), L -transposition of great arteries (n = 14 patients), tetralogy of Fallot, pulmonary atresia with ventricular septal defect (n = 11 patients), truncus arteriosus (n = 10 patients), and double-outlet ventricle (n = 3 patients). Implantation age ranged from 0.25 to 24 years (mean, 5.2 +/- 4.2 years). Median conduit diameter was 16 mm. Two-dimensional echocardiographic Doppler evaluations were made yearly; 9 patients underwent cardiac catheterization. Reintervention for stenosis was indicated when the pressure gradient exceeded 50 mm Hg. RESULTS: Three late deaths were unrelated to the conduit. Thirty-five autologous pericardial valved conduits increased in diameter (1-7 mm), remained unchanged in 15 patients, and reduced 1 to 2 mm in 4 patients. The median diameter was 18 mm at the last evaluation (P =.0001). Eight patients required conduit-related reoperation 3 to 8 years after the implantation. Two patients underwent balloon dilation of the autologous pericardial valved conduit. No conduit had to be replaced. Freedom from reintervention at 5 and 10 years was 92% and 76%, being 100% at 10 years for conduits larger than 16 mm at time of implantation. CONCLUSIONS: Autologous pericardial valved conduits show excellent long-term results and compare favorably with other conduits. PMID- 10788808 TI - Commentary PMID- 10788807 TI - Operation for partial atrioventricular septal defect: a forty-year review. AB - BACKGROUND: We describe the long-term outcome of repair of partial atrioventricular septal defect by determining the rates of survival, reoperation, and occurrence of left atrioventricular valve regurgitation, left atrioventricular valve stenosis, left ventricular outflow tract obstruction, and arrhythmia. METHODS: We studied 334 patients who underwent repair of partial atrioventricular septal defect before 1995. RESULTS: The 30-day and 5-, 10-, 20-, and 40-year survival were 98%, 94%, 93%, 87%, and 76%, respectively. Closure of the left atrioventricular valve cleft (P =. 03) and age less than 20 years at operation (P <.001) were associated with better survival. Reoperation was performed for 38 patients (11%). Repair of residual/recurrent left atrioventricular valve regurgitation or stenosis was the most common reason for reoperation. Left ventricular outflow tract obstruction occurred in 36 patients, and 7 patients underwent reoperation to relieve this obstruction. Supraventricular arrhythmias were observed in 58 patients (16%) after the operation. Supraventricular arrhythmias increased with increasing age at primary operation (P =.001). Complete atrioventricular block occurred in 9 patients (3%). Permanent pacemakers were implanted in 11 patients. CONCLUSIONS: Long-term survival after repair of partial atrioventricular septal defect is good. It is important to close the cleft in the left atrioventricular valve. Reoperation for persistent or recurrent left atrioventricular valve malfunction and relief of left ventricular outflow tract obstruction is necessary in approximately 11% of patients. PMID- 10788809 TI - Whole blood ionized magnesium: age-related differences in normal values and clinical implications of ionized hypomagnesemia in patients undergoing surgery for congenital cardiac disease. AB - OBJECTIVES: We sought to (1) determine reference values for whole blood ionized magnesium concentrations in newborns, children, and young adults and (2) evaluate the frequency and clinical implications of ionized hypomagnesemia in patients undergoing surgery for congenital heart disease. METHOD: We prospectively measured ionized magnesium concentrations in 299 subjects (113 control subjects and 186 patients undergoing surgery for congenital heart disease). Subjects were categorized by age. In the surgical group blood samples were obtained before bypass, during bypass (cooling and rewarming), after bypass, and during admission to the intensive care unit. Ionized hypomagnesemia was defined as ionized magnesium level 2 standard deviations below the mean of control subjects in the same age group. Patients were analyzed, controlling for cardiopulmonary bypass time. RESULTS: In the control group ionized magnesium concentrations differed by age. Neonates and adults showed lower ionized magnesium concentrations compared with those of other age groups. Infants exhibited the highest ionized magnesium concentration. In the surgical group patients older than 1 month showed a higher proportion of ionized hypomagnesemia compared with that found in neonates at baseline (P <.001), after bypass (P =. 03), and at admission to the intensive care unit (P =.02). Controlling for cardiopulmonary bypass time, patients older than 1 month who were hypomagnesemic during bypass showed longer intubation time (P =.001) and longer intensive care stay (P =.01) and tended to have a higher pediatric severity of illness score on intensive care admission (P =.14) compared with patients without ionized hypomagnesemia. CONCLUSIONS: There are age-related differences in normal ionized magnesium concentrations. Ionized hypomagnesemia is a common and clinically relevant occurrence among patients older than 1 month of age undergoing surgery for congenital heart disease. PMID- 10788810 TI - Detection of circulating tumor cells in patients with non-small cell lung cancer undergoing lobectomy by video-assisted thoracic surgery: a potential hazard for intraoperative hematogenous tumor cell dissemination. AB - OBJECTIVE: We prospectively tested whether circulating tumor cells can be found in the preoperative, intraoperative, and postoperative peripheral blood of patients with resectable non-small cell lung cancer who undergo video-assisted lobectomy. METHODS: We assayed for carcinoembryonic antigen messenger RNA (mRNA) by reverse transcriptase-polymerase chain reaction in the peripheral blood taken before, during, just after the completion of the lobectomy and then 2 to 3 weeks, and again 5 to 6 weeks, after the operation in 29 patients with pathologic stage I non-small cell lung cancer who underwent video-assisted lobectomy. We also analyzed the prognostic value of carcinoembryonic antigen mRNA expression pattern in an additional 57 patients with stage I non-small cell lung cancer, whose blood samples were previously assayed for carcinoembryonic antigen mRNA. RESULTS: Of the 29 patients, the preoperative blood samples from 18 patients were negative for carcinoembryonic antigen mRNA. Of these 18 patients, 16 (89%) had positive test results during operation, although the remaining 2 patients (11%) consistently showed negative test results. The occurrence of this change from negative to positive tests results for carcinoembryonic antigen mRNA during video assisted lobectomy was significantly higher than in patients who underwent open lobectomy in a previous study (18 of 35 patients; 51%; P <.001). In the 57 patients with stage I cancer whose blood samples were previously assayed for carcinoembryonic antigen mRNA, patients with persistently positive test results for carcinoembryonic antigen mRNA before and during operation had a significantly shorter survival when compared with those patients whose test results were persistently positive. CONCLUSIONS: Video-assisted lobectomy, as compared with open lobectomy, for non-small cell lung cancer may increase the risk of seeding tumor cells into the circulation during operation. PMID- 10788811 TI - Surgical treatment of pulmonary aspergilloma: current outcome. AB - OBJECTIVE: This retrospective study was designed to confirm that aggressive pulmonary resection can provide effective long-term palliation of disease for patients with pulmonary aspergilloma. METHODS AND RESULTS: From 1959 to 1998, 84 patients underwent a total of 90 operations for treatment of pulmonary aspergilloma in the Marie-Lannelongue Hospital. The mean follow-up period was 9 years, and 83% of the patients were followed up for 5 years or until death, if the latter occurred earlier. The median age was 44 years. The most common indications were hemoptysis (66%) and sputum production (15%). Fifteen patients (18%) had no symptoms. Tuberculosis and lung abscess were the most common underlying causes of lung disease (65%). The procedures were 70 lobar or segmental resections, 8 cavernostomies, and 7 pneumonectomies. Five thoracoplasties were required after lobectomy (3 patients) or pneumonectomy (2 patients). The operative mortality rate was 4%. The major complications were bleeding (23 patients), prolonged air leak (31 patients), respiratory failure (10 patients), and empyema (5 patients). The actuarial survival curve showed 84% survival at 5 years and 74% survival at 10 years. During the first 2 years, death was related to the surgical procedure and the underlying disease. In contrast, 85% of the survivors had a good late result. CONCLUSION: Lobar resection in both the symptomatic and the asymptomatic patients was conducted in low-risk settings. For patients whose condition is unfit for pulmonary resection, cavernostomy may need to be undertaken despite the high operative risk. The better survival rate in this study may have been due to the selection of patients with better lung function and localized pulmonary disease. PMID- 10788812 TI - Transforming growth factor-beta1 gene transfer ameliorates acute lung allograft rejection. AB - BACKGROUND: The aim of the current work was to study the feasibility of functional gene transfer using the gene encoding for transforming growth factor beta1, a known immunosuppressive cytokine, on rat lung allograft function in the setting of acute rejection. METHODS: The rat left lung transplant technique was used in all experiments, with Brown Norway donor rats and Fischer recipient rats. After harvest, left lungs were transfected ex vivo with either sense or antisense transforming growth factor-beta1 constructs complexed to cationic lipids, then implanted into recipients. On postoperative days 2, 5, and 7, animals were put to death, arterial oxygenation measured, and acute rejection graded histologically. RESULTS: On postoperative day 2, there were no differences in acute rejection or lung function between animals treated with transforming growth factor-beta1 and control animals. On postoperative day 5, oxygenation was significantly improved in grafts transfected with the transforming growth factor-beta1 sense construct compared with antisense controls (arterial oxygen tension = 411 +/- 198 vs 103 +/ 85 mm Hg, respectively; P =.002). Acute rejection scores from lung allografts were also significantly improved, corresponding to decreases in both vascular and airway rejection (vascular rejection scores: 2.0 +/- 0. 5 vs 2.8 +/- 0.6; P =.04; airway rejection scores: 1.3 +/- 0.7 vs 2. 3 +/- 0.8, respectively; P =.02). The amelioration of acute rejection was temporary and decreased by postoperative day 7. CONCLUSIONS: The feasibility of using gene transfer techniques to introduce novel functional genes in the setting of lung transplantation is demonstrated. In this model of rat lung allograft rejection, gene transfer of transforming growth factor-beta1 resulted in temporary but significant improvements in lung allograft function and acute rejection pathology. PMID- 10788814 TI - Acute hypoxia and reoxygenation impairs exhaled nitric oxide release and pulmonary mechanics. AB - OBJECTIVE: Changes in exhaled nitric oxide levels often accompany conditions associated with elevated pulmonary vascular resistance and altered lung mechanics. However, it is unclear whether changes in exhaled nitric oxide reflect altered vascular or bronchial nitric oxide production. This study determined the effects of acute hypoxia and reoxygenation on pulmonary mechanics, plasma nitrite levels, and exhaled nitric oxide production. METHODS: Ten piglets underwent 90 minutes of hypoxia (fraction of inspired oxygen = 12%), 1 hour of reoxygenation on cardiopulmonary bypass, and 2 hours of recovery. Five additional animals underwent bypass without hypoxia. Exhaled nitric oxide, plasma nitrite levels, and pulmonary mechanics were measured. RESULTS: Exhaled nitric oxide decreased to 36% of baseline by end hypoxia (34 +/- 14 vs 12 +/- 9 ppb, P =.005) and declined further to 20% of baseline at end recovery (7 +/- 6 ppb). Aortic nitrite levels decreased from baseline during hypoxia (from 102 +/- 13 to 49 +/- 7 micromol/L, P =.05) but returned to baseline during recovery. Pulmonary arterial nitrite also decreased during hypoxia (from 31.4 +/- 7.8 to 22.9 +/- 9.5 micromol/L, P =.04) and returned to baseline at end recovery. Decreased production of exhaled nitric oxide was associated with impaired gas exchange (alveolar-arterial gradient = 32 mm Hg at baseline and 84 mm Hg at end recovery), decreased pulmonary compliance (6.6 +/- 0.9 mL/cm H(2)O at baseline, 5.0 +/- 0.7 mL/cm H(2)O at end hypoxia, and 5.4 +/- 0.7 mL/cm H(2)O at end recovery), and increased inspiratory airway resistance (41 +/- 4 cm H(2)O. L(-1). s(-1) at baseline, 56 +/- 4.9 cm H(2)O. L( 1). s(-1) at end hypoxia, and 50 +/- 5 cm H(2)O. L(-1). s(-1) at end recovery). CONCLUSIONS: A decrease in exhaled nitric oxide persisted after hypoxia, and plasma nitrite levels returned to baseline on reoxygenation, indicating that alterations in exhaled nitric oxide during hypoxia-reoxygenation might be unrelated to plasma nitrite levels. Furthermore, decreased exhaled nitric oxide corresponded with altered pulmonary mechanics and gas exchange. Reduced exhaled nitric oxide after hypoxia-reoxygenation might reflect bronchial epithelial dysfunction associated with acute lung injury. PMID- 10788813 TI - Comparison of university of wisconsin, euro-collins, low-potassium dextran, and krebs-henseleit solutions for hypothermic lung preservation. AB - OBJECTIVE: We sought to test the effectiveness of 4 different solutions for hypothermic rat lung preservation. METHODS: One hundred ninety-two rats were used. The rats were divided into 4 groups, and University of Wisconsin, Euro Collins, low-potassium dextran, or Krebs-Henseleit solution was used in each group. They were further divided into 6 subgroups of 8 rats each. The lungs were preserved at 4 degrees C for 0, 4, 6, 8, 12, or 24 hours, respectively, and lung function was studied by using a living rat perfusion model. RESULTS: Pulmonary arterial flow decreased in each group after 4 to 6 hours of preservation; the low potassium dextran group decreased the least and the Krebs-Henseleit group decreased the most. Pulmonary vascular resistance increased in each group after 6 hours of preservation; the Krebs-Henseleit group increased the most. Although airway pressure increased, static lung compliance and gas exchange capacity decreased after 8 hours of preservation; the Krebs-Henseleit group exhibited the worst values. Lung tissue wet/dry weight ratio increased gradually during preservation; the University of Wisconsin group exhibited the least increase. An ultrastructural study indicated the least morphologic changes in the low potassium dextran group at 24 hours. CONCLUSIONS: At 4 degrees C, all solutions preserved rat lungs for 4 hours with acceptable function. However, 6 hours of preservation resulted in damaged pulmonary function in some lungs, and this damage increased when preservation time was extended. The lungs preserved in low potassium dextran solution had the best overall function, but the lungs preserved in University of Wisconsin solution had less edema. PMID- 10788815 TI - Effects of preoperative chemotherapy and radiation therapy on human bronchial blood flow. AB - OBJECTIVE: We investigated the relationship between bronchial mucosal blood flow around the area of lung resection and the state of healing of the bronchial stump in patients after chemotherapy with or without radiation therapy. METHODS: Ninety patients with primary lung cancer were divided into the following 3 groups: group A, 72 patients who had no preoperative therapy; group B, 10 patients who had chemotherapy; and group C, 8 patients who had chemoradiation (60 Gy) therapy. Bronchial mucosal blood flow was measured preoperatively, intraoperatively, and postoperatively (days 8-10) with a laser Doppler flowmeter. RESULTS: In groups A and B bronchial mucosal blood flow was preserved sufficiently around the surgical site, and the healing of the bronchial stump was satisfactory. On the contrary, preoperative blood flow in group C was 70% of the preoperative value in group A and decreased further intraoperatively. Healing of the bronchial stump was poor, and a bronchopleural fistula occurred in one patient of group C. CONCLUSION: Preoperative chemoradiation therapy may adversely affect bronchial mucosal blood flow and healing of the bronchial stump, although lymphadenectomy and preoperative chemotherapy had little effect. It is recommended that the bronchial stump should be covered with pedicled viable tissue after chemoradiation therapy for prophylaxis against bronchial complications. PMID- 10788816 TI - Long-term effectiveness of operations for ascending aortic dissections. AB - OBJECTIVE: To evaluate long-term effectiveness of a strategy for managing the aortic root and distal aorta according to the pathology in ascending aortic dissection. METHODS: From 1978 to 1995, 208 patients underwent operations for acute (n = 135) and chronic (n = 73) ascending aortic dissection. Surgical strategies included valve resuspension with supracoronary aortic root repair and ascending aortic graft for normal sinuses and valve (n = 135), composite valve and ascending aortic graft for abnormal sinuses and valve (n = 47), and valve replacement and supracoronary ascending aortic graft for normal sinuses and abnormal valve (n = 26). Resection extended into the arch only if the intimal tear originated in or extended to the aortic arch (n = 31). RESULTS: Hospital mortality was 14%. Cardiogenic shock (P =.002) and concomitant coronary artery bypass grafting (P =.001) were associated with increased risk; use of circulatory arrest (P =.0003) decreased risk. Survival was 87%, 68%, and 52% at 30 days, 5 years, and 10 years, respectively. Advanced age, earlier date of operation, composite graft, and arch resection were associated with decreased survival; residual distal dissected aorta was not. Reoperation was required for 5 proximal and 8 distal problems. CONCLUSIONS: In both acute and chronic ascending aortic dissections, (1) circulatory arrest is associated with low early mortality; (2) with normal sinuses and valve, supracoronary repair of the dissected aortic root and valve resuspension is effective long term; and (3) residual distal dissected aorta does not decrease late survival and has a low risk of aneurysmal change and reoperation for at least 10 years. PMID- 10788817 TI - Aortic valve replacement: is valve size important? AB - OBJECTIVE: We sought to determine whether aortic prosthesis size adversely influences survival after aortic valve replacement. METHODS: A total of 892 adults receiving a mechanical (n = 346), pericardial (n = 463), or allograft (n = 83) valve for aortic stenosis were observed for up to 20 years (mean, 5.0 +/- 3.9 years) after primary isolated aortic valve replacement. We used multivariable propensity scores to adjust for valve selection factors, multivariable hazard function analyses to identify risk factors for all-cause mortality, and bootstrap resampling to quantify the reliability of the results. RESULTS: Twenty-five percent of patients had indexed internal orifice areas of less than 1.5 cm(2)/m(2) and more than 2 SDs (Z-value) below predicted normal aortic valve size. Mechanical valve orifices were smaller (1.3 +/- 0. 29 cm(2)/m(2), Z = -2.2 +/- 1.16) than pericardial (1.9 +/- 0.36 cm(2)/m(2), Z = -0.40 +/- 1.01) or allograft valves (2.1 +/- 0.50, Z = 0.24 +/- 1.17). The overall survival was 98%, 96%, 86%, 69%, and 49% at 30 days and 1, 5, 10, and 15 years postoperatively. Univariably, survival was weakly and inversely related to manufacturer valve size (P =.16) and internal orifice diameter (P =. 2) but completely unrelated to indexed valve area (P =.6) or Z-value (P =.8). These, and univariable differences among valve types (P =. 004), were accounted for by different prevalences in patient risk factors and not by valve size or type per se. Bootstrap resampling indicated that these findings had a less than 15% chance of being incorrect. CONCLUSIONS: Survival after aortic valve replacement is strongly related to patient risk factors but appears not to be adversely affected by moderate patient prosthesis mismatch (down to about 4 SDs below normal). Aortic root enlargement to accommodate a large prosthesis may be required in few situations. PMID- 10788818 TI - Body surface area as a predictor of aortic and pulmonary valve diameter. AB - BACKGROUND: Predicting cardiac valve size from noncardiac anatomic measurements would benefit pediatric cardiologists, adult cardiologists, and cardiac surgeons in a number of decision-making situations. Previous studies correlating valve size with body size have been generated with the use of fixed autopsy specimens, angiography, and echocardiography, but primarily in the young. This study examines the relation of body surface area to measurements of the left ventricular-aortic junction (aortic valve anulus diameter) and the right ventricular-pulmonary trunk junction (pulmonary valve anulus diameter) in 6801 hearts across a wide spectrum of ages. METHODS: From June 1985 to October 1998, cardiac valves from 6801 donated hearts were analyzed morphologically. Donor age was newborn to 59 years (mean 31 +/- 17 years; median 32 years). Calculated body surface areas ranged from 0.18 to 3.55 m(2). Aortic (n = 4636) and pulmonary valve diameters (n = 5480) were measured from enucleated valves suitable for allograft transplantation. Mean valve sizes were computed for ranges in body surface area in 0.1-m(2) increments. RESULTS: For adult men (age >/= 17 years), the mean aortic valve diameter was 23.1 +/- 2.0 mm (n = 2214) and the mean pulmonary valve diameter was 26.2 +/- 2.3 mm (n = 2589). For adult women, the mean aortic valve diameter was 21.0 +/- 1.8 mm (n = 1156) and the mean pulmonary valve diameter was 23.9 +/- 2.2 mm (n = 1408). The mean indexed aortic valve area was 2.02 +/- 0.52 cm(2)/m(2) and the pulmonary valve area 2.65 +/- 0.52 cm(2)/m(2). Between 82% and 85% of the variability was explained by the size of the patient. Regression equations were developed both overall and separately for men and women, although the additional contribution of sex above that of body size was less than 1%. CONCLUSIONS: Aortic and pulmonary valve diameters are closely related to body size. Thus, body surface area, when used in conjunction with other clinically accepted evaluations, is a useful tool for estimating normal aortic and pulmonary valve size. PMID- 10788819 TI - Inducible expression of basic transcription element-binding protein 2 in proliferating smooth muscle cells at the vascular anastomotic stricture. AB - OBJECTIVE: The proliferation of vascular smooth muscle cells surrounding a suture line is an important factor in the development of anastomotic stricture that is frequently seen after coronary artery bypass grafting. The aim of this study was to investigate the time course of intimal thickening and to examine the expression of the molecular marker of smooth muscle cell activation surrounding the suture line. METHODS: Longitudinal aortotomy was performed in the abdominal aorta of rats. The rats were put to death 1, 2, 4, and 8 weeks after aortotomy, and the percentage of the lumen occluded by intimal thickening was calculated. All tissues were stained with antibodies against basic transcription element- binding protein 2, human cyclin-dependent kinase (cdk4), and Sp1 for immunohistochemistry. Basic transcription element-binding protein 2 is a transcription factor that is involved in phenotypic modulation of vascular smooth muscle cells. Cdk4 represents a marker for G(1) phase of the cell cycle. Sp1 is a transcription factor known to be expressed in a variety of tissues. Basic transcription element-binding protein 2 messenger RNA expression was confirmed by means of reverse transcriptase-polymerase chain reaction. RESULTS: We noted significant thickening of the intimal layer 1 week after aortotomy. Immunohistochemistry demonstrated that smooth muscle cells in the neointima were strongly positive for basic transcription element-binding protein 2 and human cyclin-dependent kinase 4, which peaked 2 weeks after aortotomy. Basic transcription element-binding protein 2 expression was closely associated with human cyclin-dependent kinase 4 expression in the neointima, although Sp1 was not. Basic transcription element-binding protein 2 messenger RNA levels were significantly up-regulated early after aortotomy. CONCLUSION: The experimental rat aortotomy model is useful to investigate the proliferation of vascular smooth muscle cells around the suture line. Moreover, our results suggest the possible role of basic transcription element-binding protein 2 in the development of vascular anastomotic strictures. PMID- 10788820 TI - Size and distensibility of the aortic root and aortic valve function after different techniques of the ross procedure. AB - OBJECTIVES: In the Ross procedure, 3 different techniques are used for aortic valve replacement with the pulmonary autograft: freestanding root, inclusion, and subcoronary implantation. The objective of this study was to evaluate echocardiographically the influence of the particular operative technique on dimension, distensibility, and valve function. METHODS: Between February 1990 and August 1998, the Ross procedure was performed in 111 patients (mean age, 48.6 +/- 14.1 years; range, 15.2-70.6 years), with 1 early and 1 late death, 1 autograft replacement, and 1 patient lost to follow-up. The remaining patients underwent the freestanding root (n = 9 patients), inclusion (n = 14 patients), and subcoronary techniques (n = 84 patients). Echocardiography was performed at a mean follow-up of 26 +/- 21.3 months after operation and was compared with the echocardiographic findings of the control subjects (n = 10 subjects). Root sizes were measured at the level of the anulus, sinus, and supra-aortic ridge; the distensibility was calculated as pressure strain elastic modulus and percent change of radius. RESULTS: Size and distensibility of the aortic root were normal, except for a larger diameter at the sinus level in the root technique in comparison to the subcoronary technique (P <.05; maximum diameter, 41.3 +/- 8.6 mm vs 32.6 +/- 4.0 mm). Aortic valve function was comparable among groups with low pressure gradients and most patients with no or trace aortic insufficiency. CONCLUSIONS: The freestanding root, inclusion, and subcoronary techniques in the Ross procedure provide comparable excellent hemodynamics, normal root size, and distensibility, except for the enlarged sinus diameter in the freestanding root. These results may have some impact on the operative procedure and follow-up investigations. PMID- 10788821 TI - Potential role of vacuolar H-adenosine triphosphatase in neointimal formation in cultured human saphenous vein. AB - OBJECTIVE: Vacuolar H(+)-adenosine triphosphatase plays a pivotal role in pH regulation and molecular transport across the vacuolar membranes and is involved in cell proliferation and transformation. In the present study, possible involvement of vacuolar H(+)-adenosine triphosphatase in neointimal formation was investigated in an organ culture model of human saphenous vein. METHODS AND RESULTS: Cultured saphenous vein segments developed neointimal formation and marked thickening of the media within 14 days. Neointimal formation and medial thickening were completely inhibited by 10 nmol/L bafilomycin A(1), a selective inhibitor of vacuolar H(+)-adenosine triphosphatase, although structurally related macrolide antibiotics FK-506 and erythromycin were without an effect. The neointimal cells were positive for alpha-smooth muscle actin and vimentin but negative for desmin, indicative of myofibroblasts. The emergence of myofibroblasts was inhibited, and endothelial cells were preserved in the saphenous vein segments treated with bafilomycin A(1). Uptake of bromodeoxyuridine, a proliferation marker, by myofibroblasts was abrogated in the saphenous vein segments treated with 10 nmol/L bafilomycin A(1). Detection of apoptotic cells by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling concomitant with identification of desmin-expressing smooth muscle cells demonstrated that neointimal myofibroblasts, but not medial smooth muscle cells, that expressed desmin underwent apoptosis by treatment with bafilomycin A(1). CONCLUSIONS: These results suggest that vacuolar H(+)-adenosine triphosphatase may be involved in myofibroblast growth that contributes to neointimal formation and medial thickening in cultured human saphenous vein. Increased sensitivity of myofibroblasts, but not endothelial cells, and differentiated smooth muscle cells to bafilomycin A(1) may have potential therapeutic implications in the treatment for vein graft disease. PMID- 10788823 TI - Commentary PMID- 10788822 TI - Coronary artery bypass surgery with arterial grafts in familial hypercholesterolemia. AB - OBJECTIVE: Familial hypercholesterolemia is a dominantly inherited disorder caused by mutations at the locus for the low-density lipoprotein receptor and is frequently associated with premature coronary artery disease. This study was performed to determine whether arterial grafting was associated with long-term benefits for patients with familial hypercholesterolemia. METHODS: During the past 18 years, 101 patients with heterozygous familial hypercholesterolemia underwent primary coronary artery bypass grafting, with one hospital death. Group 1 patients (n = 31) received only saphenous vein grafts. Group 2A patients (n = 47) received one internal thoracic artery graft and supplemental vein grafts, and group 2B patients (n = 23) had multiple arterial grafts. After operation, all patients received diet therapy and intensive cholesterol-lowering drug therapy. Thirteen patients received low-density lipoprotein apheresis. RESULTS: During a mean follow-up period of 95 months, 8 patients died, 9 underwent reoperation, and 12 received catheter intervention. The overall survival was 82% (95% confidence limits, 65%-97%) at 18 years after operation. The survival in group 2 was higher than that found in group 1 (P =.01). The overall freedom from major cardiac events (myocardial infarction, cardiac death, reoperation, and catheter intervention) was 57% (95% confidence limits, 40%-74%) at 16 years after operation. The freedom from reoperation in group 2 was higher than that found in group 1 (P =.03). There was no difference in the survival or freedom from major cardiac events between groups 2A and 2B. CONCLUSION: Arterial grafting improved the long-term freedom from reoperation in patients with familial hypercholesterolemia. Additional benefit of multiple arterial grafting could not be identified. PMID- 10788824 TI - Transfer of patients receiving advanced mechanical circulatory support. AB - OBJECTIVE: Improving results with ventricular assist devices have led to their wider clinical application. Centers can stabilize, support, and wean or transfer patients to regional transplant centers. METHODS: Prospectively collected data were reviewed to evaluate the clinical results of patients transferred to our institution while receiving advanced mechanical circulatory support. RESULTS: Since 1993, 16 patients were accepted while receiving support with either extracorporeal membrane oxygenation or a ventricular assist device. The 10 male and 6 female patients ranged in age from 9 to 60 years (mean, 42.1 years). Thirteen had had cardiac surgical procedures, two had acute myocardial infarctions, and one had myocarditis. The distance transported ranged from 0.2 to 309 miles (mean, 132 miles). Twelve patients were transferred by ground, and 4 were transported by air. Seven patients were originally supported with extracorporeal membrane oxygenation, 6 with centrifugal pumps, and 3 with ABIOMED ventricular assist devices (ABIOMED, Inc, Danvers, Mass). Two patients had clinical complications during transfer, and one had a cerebrovascular accident, recovered, was weaned, and survived. A second patient had hemodynamic deterioration. There were no technical complications associated with transport. Six patients were left on the original support device; 3 of the 6 were weaned and survived, and 3 died during support. The 10 remaining patients were switched to other ventricular assist devices: 9 patients to Thoratec devices (Thoratec Laboratories, Pleasanton, Calif) and 1 patient to a Novacor device (Baxter Healthcare Corp, Novacor Division, Oakland, Calif). Six of the 10 patients underwent transplantation and survived. Four patients died while being supported by the devices. Nine patients were discharged, with 1 late death at 29 months. Eight patients are alive 4 to 65 months after discharge. CONCLUSIONS: These data suggest that patients receiving advanced support can be moved between clinical centers with acceptable risks. Because 33% of the survivors were weaned, transplantation is not required for survival. PMID- 10788825 TI - Is maintained cranial hypothermia the only factor leading to improved outcome after retrograde cerebral perfusion? An experimental study with a chronic porcine model. AB - BACKGROUND: Previous studies have shown that retrograde cerebral perfusion can improve neurologic outcome after prolonged hypothermic circulatory arrest. Here we have compared two temperatures of retrograde cerebral perfusion (15 degrees C and 25 degrees C) with hypothermic circulatory arrest at systemic hypothermia of 25 degrees C to clarify whether the possible benefit of retrograde cerebral perfusion may only be due to improved cooling effect. METHODS: Eighteen pigs (23 27 kg) were randomly assigned to undergo 15 degrees C retrograde cerebral perfusion at systemic hypothermia of 25 degrees C, 25 degrees C retrograde cerebral perfusion at 25 degrees C systemic hypothermia, or hypothermic circulatory arrest at 25 degrees C for 40 minutes. Flow was adjusted to maintain superior vena cava pressure at 20 mm Hg during retrograde cerebral perfusion. Hemodynamic, electrophysiologic, metabolic, and temperature monitoring were performed until 4 hours after the start of rewarming. Daily behavioral assessment was done until death or until the animals were killed on day 7. Histopathologic analysis of the brain was carried out on all animals. RESULTS: Epidural temperatures were lower in the 15 degrees C retrograde cerebral perfusion group during the intervention (P <.05). In the 15 degrees C retrograde cerebral perfusion group, 4 (67%) of 6 animals survived for 7 days compared with 3 (50%) of 6 in both the 25 degrees C retrograde cerebral perfusion and hypothermic circulatory arrest groups. The median total histopathologic score was 5 in the 15 degrees C retrograde cerebral perfusion group and 7 in the 25 degrees C retrograde cerebral perfusion group (P =.04). CONCLUSIONS: These findings suggest that enhanced cranial hypothermia is the major beneficial factor of retrograde cerebral perfusion when careful attention is paid to its implementation. PMID- 10788826 TI - Captopril-induced glutamate release at the start of reperfusion after cold cardioplegic storage of pig hearts. AB - OBJECTIVE: We sought to evaluate the effects of captopril on glucose-related metabolism during hypothermic cardioplegic storage and subsequent reperfusion. METHODS: We compared hearts from control pigs with hearts from pigs treated with increasing oral doses of captopril for 3 weeks (12.5-150 mg daily), an intravenous bolus (25 mg) before operation, and captopril-containing cardioplegic solution (1 mg/L). The hearts were excised after infusion of cold crystalloid cardioplegic solution and stored in saline solution (4 degrees C-6 degrees C). In one series we studied myocardial blood flow and arteriovenous differences in oxygen, glucose, lactate, glutamate, and alanine during 60 minutes of postcardioplegic blood reperfusion. In this series captopril-treated hearts were reperfused with captopril-containing blood (1 mg/L). In another series we obtained biopsy specimens from the left ventricle throughout 30 hours of hypothermic cardioplegic storage and monitored tissue content of energy-rich phosphates, glycogen, glutamate, and alanine. RESULTS: Captopril increased glutamate and alanine release 11- to 17-fold at the start of reperfusion (P <.001). Furthermore, captopril increased myocardial oxygen and glucose uptake during reperfusion (P <.001 for both), whereas lactate release and myocardial blood flow were unaffected by captopril. At the start of reperfusion, there was a positive correlation between glutamate release and glucose uptake in captopril treated hearts (r = 0.66, P =.05). We found no statistically significant differences between captopril and control hearts in tissue content of adenosine triphosphate, glycogen, glutamate, alanine, or lactate during 30 hours of cardioplegic storage. CONCLUSIONS: The metabolic effects of captopril are strictly related to reperfusion, during which oxidative metabolism of glucose is improved. The captopril-induced increase in glutamate and alanine release at the start of reperfusion after cardioplegic storage may reflect a switch in metabolism of glucose-related amino acids. PMID- 10788828 TI - Spontaneous anterior thoracic lung hernias. PMID- 10788827 TI - Vasorelaxant effect of phosphodiesterase-inhibitor milrinone in the human radial artery used as coronary bypass graft. AB - OBJECTIVE: The radial artery is a spastic coronary bypass graft. We investigated the effect of the phosphodiesterase III inhibitor milrinone on the human radial artery. METHODS: Radial artery segments (n = 76) taken from 15 patients were studied in an organ chamber. Concentration-relaxation curves for milrinone were established in the radial artery precontracted with 3 vasoconstrictors (phenylephrine, K(+), and U46619). In radial artery rings incubated with therapeutic plasma concentrations of milrinone (7 and 70 micromol/L) for 10 minutes, concentration-contraction curves for the 3 vasoconstrictors were constructed. RESULTS: Milrinone caused a submaximal relaxation in phenylephrine- (98.6% +/- 1.4%), K(+)- (89.1 +/- 4.5%), or U46619- (74.2 +/- 8.0%) precontracted radial arteries at -4.5 log(10) M. The EC(50) was higher against K(+) (-5.85 +/- 0.24 log(10) M, P =.02) or U46619 (-5. 21 +/- 0.61 log(10) M, P =.03) than phenylephrine (-6.68 +/- 0.11 log(10) M). Pretreatment with milrinone depressed the contraction by phenylephrine from 70.0% +/- 7.9% to 23.5% +/- 9.3% (P =.003) and by K(+) from 138.6% +/- 5.8% to 73.0% +/- 13.9% (P =.006) and shifted the EC(50) 3.8-fold higher (P =.03) for phenylephrine and 2.2-fold higher for K(+) (P =.01). Milrinone reduced the U46619 contraction at low concentration (-8.5 log(10) M) but had little effect on the maximal contraction. CONCLUSION: Milrinone is a potent vasodilator for the radial artery, with possibly higher potency in alpha-adrenoceptor- and depolarizing agent K(+)-mediated, but less potency in thromboxane A(2)-mediated, contraction. Because it also has a positive inotropic effect, this vasodilator may be particularly indicated for use in patients receiving radial artery grafts in coronary artery bypass grafting. PMID- 10788829 TI - A new technique for sparing the aortic valve in patients with aneurysm of the ascending aorta and root. PMID- 10788830 TI - Reversal of spinal cord ischemia resulting from aortic dissection. PMID- 10788831 TI - Macro design, structure, and mechanics of the left ventricle. PMID- 10788832 TI - Warm reperfusion as an adjunct to myocardial protection. PMID- 10788833 TI - Brain injury in preterm newborn infants. PMID- 10788835 TI - Noonan syndrome: a clinical and genetic study of 31 patients. AB - Noonan syndrome is a multiple congenital anomaly syndrome, inherited in an autosomal dominant pattern. We studied 31 patients (18 males and 13 females) affected by this disorder regarding their clinical and genetic characteristics. The most frequent clinical findings were short stature (71%); craniofacial dysmorphisms, especially hypertelorism, ptosis, downslanting of the palpebral fissures; short or webbed neck (87%); cardiac anomalies (65%), and fetal pads in fingers and toes (70%). After studying the probands' first-degree relatives, we made the diagnosis of Noonan syndrome in more than one family member in three families. Therefore, the majority of our cases were sporadic. PMID- 10788834 TI - Life cycle of Trypanosoma cruzi (Y strain) in mice. AB - Since 1958, we have studied experimental Chagas' disease (CD) by subcutaneous inoculation of 1,000 blood forms of Trypanosoma cruzi (Y strain) in Balb/C. mice. Evolution of parasitemia remained constant, beginning on the 5th and 6th day of the disease, increasing progressively, achieving a maximum on about the 30th day. After another month, only a few forms were present, and they disappeared from the circulation after the third month, as determined from direct examination of slides and the use of a Neubauer Counting Chamber. These events coincided with the appearance of amastigote nests in the tissues (especially the cardiac ones), starting the first week, and following the Gauss parasitemia curve, but they were not in parallel until the chronic stage. In 1997, we began to note the following changes: Parasites appeared in the circulation during the first week and disappeared starting on the 7th day, and there was a coincident absence of the amastigote nests in the tissues. A careful study verified that young forms in the evolutionary cycle of T. cruzi (epi + amastigotes) began to appear alongside the trypomastigotes in the circulation on the 5th and 7th post-inoculation day. At the same time, rounded, oval, and spindle shapes were seen circulating through the capillaries and sinusoids of the tissues, principally of the hematopoietic organs. Stasis occurs because the diameter of the circulating parasites is greater than the vessels, and this makes them more visible. Examination of the sternal bone marrow revealed young cells with elongated forms and others truncated in the shape of a "C" occupying the internal surface of the blood cells that had empty central portions (erythrocytes?). We hypothesize that there could be a loss of virulence or mutation of the Y strain of Trypanosoma cruzi. PMID- 10788836 TI - Intraventricular hemorrhage in very low birth weight infants: associated risk factors and outcome in the neonatal period. AB - Intraventricular hemorrhage (IVH) is a severe complication in very low birth weight (VLBW) newborns (NB). With the purpose of studying the incidence of IVH, the associated risk factors, and the outcomes for these neonates, we studied all the VLBW infants born in our neonatal unit. Birth weight, gestational age, presence of perinatal asphyxia, mechanical ventilation, length of hospitalization, apnea crisis, hydrocephalus, and periventricular leukomalacia were analyzed. The diagnosis of IVH was based on ultrasound scan studies (Papile's classification) performed until the tenth day of life and repeated weekly in the presence of abnormalities. Sixty-seven/101 neonates were studied. The mortality rate was 30.6% (31/101) and the incidence of IVH was 29.8% (20/67) : 70% grade I, 20% grade III and 10% grade IV. The incidence of IVH in NB <1,000 g was 53.8% (p = 0. 035) and for gestational age <30 weeks was 47.3% (p = 0.04), both considered risk factors for IVH. The length of hospitalization (p = 0.00015) and mechanical ventilation (p = 0.038) were longer in IHV NB. The IVH NB had a relative risk of 2.3 of developing apnea (p = 0. 02), 3.7 of hydrocephalus (p = 0.0007), and 7.7 of periventricular leukomalacia (p < 0.00001). The authors emphasize the importance of knowing the risk factors related to IVH so as to introduce prevention schemes to reduce IVH and to improve outcomes of affected newborns. PMID- 10788837 TI - Pouchitis: extracolonic manifestation of ulcerative colitis? AB - Pouchitis is the most frequent complication of ileal pouch-anal anastomosis for treatment of ulcerative colitis. There are several possible explanations. Among them, we focus on the one that considers pouchitis as an extracolonic manifestation of ulcerative colitis. The aim of this study was to investigate the association between pouchitis and extra-intestinal manifestations (EIM), which are frequent in these patients. Sixty patients underwent restorative proctocolectomy with an ileal J pouch (IPAA) from September 1984 to December 1998. Pouchitis was defined by clinical, endoscopic, and histologic criteria. The following extra-intestinal manifestations were studied: articular, cutaneous, hepatobiliary, ocular, genitourinary, and growth failure. Thirteen patients, of which 10 were female (76.9%), developed one or more episodes of pouchitis. Twelve patients of this group (92.3%) presented some kind of extra-intestinal manifestation, 4 pre-operatively (exclusively), 2 post-operatively (exclusively), and 6 both pre- and post-operatively (1.7 per patient). Twenty patients (42.7%) of the 47 without pouchitis did not present extra-intestinal manifestations; 10/35 (28. 5%) of females had pouchitis, compared to 3/35 (12.0%) of men. Pouchitis was more frequent among females, though not statistically significant. EIM increases the risk of pouchitis. Pouchitis is related to EIM in 92.3 % of cases, corroborating the hypothesis that it could be an extracolonic manifestation of ulcerative colitis. PMID- 10788838 TI - Unusual abdominal tumors with intracardiac extension. Two cases with successful surgical resection. AB - Abdominal tumors that can grow through vascular lumen and spread to the right heart are rare. Although these tumors have different histologic aspects, they may cause similar abdominal and cardiac symptoms and are a serious risk factor for pulmonary embolism and sudden death when they reach the right atrium and tricuspid valve. The best treatment is radical surgical resection of the entire tumor using cardiopulmonary bypass with or without deep hypothermia and total circulatory arrest. We report the cases of two patients, the first with leiomyosarcoma of the inferior vena cava and the other with intravenous leiomyomatosis of the uterus that showed intravascular growth up to right atrium and ventricle, who underwent successful radical resection in a one-stage procedure with the use of cardiopulmonary bypass. We discuss the clinical and histologic aspects and imaging diagnosis and review the literature. PMID- 10788839 TI - Primary biliary cirrhosis and myopathy: an uncommon association. AB - Primary biliary cirrhosis (PBC) is a cholestatic liver disease, which is characterized by a chronic inflammatory destruction of intrahepatic bile ducts. It is a rare disorder whose precise etiology is still to be elucidated. Even though the liver is the principal target of PBC, other organ systems also might be affected. Muscular involvement has rarely been described in this disease, and in the majority of cases, muscular weakness has been interpreted as polymyositis. We report the case of a 48-year-old woman suffering from classic PBC, in association with a myopathy whose histological features are distinct from the cases reported before. We also performed a MEDLINE research for PBC and concomitant muscular diseases. PMID- 10788840 TI - Mycetoma in an HIV-infected patient. AB - Although opportunistic fungal infections occur commonly in immunocompromised hosts, mycetoma has never been reported in association with HIV infection. The authors present a case that to their knowledge is the first reported case of mycetoma associated with HIV infection. Diagnosis was confirmed by direct examination of grains and histologic examination. Precise identification of the agent, an actinomycete, was not possible. The unusual site of infection may probably be related to the use of contaminated needless and syringes for HIV drug injection. PMID- 10788841 TI - Activity of macrolides, lincosamines, streptogramins and fluoroquinolones against streptococcus pneumoniae and enterococci isolates from the western hemisphere: example of international surveillance (SENTRY antimicrobial surveillance program )in the development of new drugs. AB - Resistance among commonly isolated Gram-positive cocci have compromised the available therapeutic regimens and require structured monitoring at the local, regional, national, and international levels. Two popular treatment classes of antimicrobials (macrolides-lincosamines-streptogramins [MLS], fluoroquinolones) have been tested against 3, 049 isolates of Streptococcus pneumoniae and enterococci from the SENTRY Antimicrobial Surveillance program. The strains were obtained from clinical cases in hospitals in the United States, Canada, and six nations (10 medical centers )in Latin America. MLS and fluoroquinolone compounds had moderate activity against vancomycin-susceptible Enterococcus faecalis only (gatifloxacin, and trovafloxacin MIC(50), 0.5 microg/ml), and quinupristin/dalfopristin was potent only against E.faecium isolates (MIC(90), 1 microg/ml(-2) microg/ml). When tested against pneumococci, gatifloxacin, trovafloxacin, sparfloxacin, and quinupristin/dalfopristin (MIC(90), < or = 1 microg/ml)were most active among the newer drugs, but vancomycin and clindamycinn inhibited > or =99.8% and 84.7% to 99.1% of strains, respectively. These results from a global resistance monitoring program should encourage rapid drug development. Based on in vitro sensitivity testing, they indicate a promising role for the treatment of emerging resistant Gram-positive cocci. The clinical role for each new agent will depend on safety profiles, rates of administration, and other issues identified during development in the clinical trials process. PMID- 10788842 TI - Comparative evaluation of the in vitro activity of three combinations of beta lactams with beta-lactamase inhibitors: piperacillin/tazobactam, ticarcillin/clavulanic acid and ampicillin/sulbactam. AB - Recently, two new combinations of Beta-lactam antibiotics with Beta-lactamase inhibitors became commercially available in Brazil: piperacillin/tazobactam and ampicillin/sulbactam. This study was designed to assess and compare the in-vitro activity of these new compounds, as well as that of ticarcillin/clavulanic acid, against bacteria isolated in our environment. A total of 749 bacteria isolated at Sao Paulo Hospital were tested using the disk diffusion method, in compliance with NCCLS standardization, using strict quality control. Only one sample per patient was included in the study. Oxacillin-resistant staphylococcus samples were not included in this study. Of the total samples tested, 84.5% were susceptible to piperacillin/tazobactam, 81.2% to ticarcillin/clavulanic acid, and 77.6% to ampicillin/sulbactam. Piperacillin/tazobactam was also found to be the most active combination of the three against Enterobacteriaceae ( n = 312), inhibiting 91.7% of the bacteria tested. Ticarcillin/clavulanic acid was active against 85.8% of the Enterobacteriaceae, while ampicillin/sulbactam inhibited 83.2% of the samples. This order of the spectrum of action (piperacillin/tazobactam > ticarcillin/clavulanic acid >ampicillin/sulbactam )was maintained for the majority of Enterobacteriaceae species analyzed. Pseudomonas aeruginosa ( n = 117) showed extremely high resistance to the three combinations. Piperacillin/tazobactam was active against 61.5% of the samples, while ticarcillin/clavulanic acid was active against 56.4% of the samples of this species. The activity of ampicillin/sulbactam against P. aeruginosa was extremely low; however, this was the most active combination against Acinetobacter baumannii ( 87.0% susceptibility). Piperacillin/tazobactam was the most active combination against Stenotrophomonas (Xanthomonas )maltophilia (100% susceptibility) and Burkholderia cepacia (90.9% susceptibility). The three combinations showed excellent activity against the Gram-positive cocci tested (97.3% to 98.2% susceptibility). In sum, piperacillin/tazobactam was more active against all Gram-negative species than the other two combinations, with the exception of A. baumannii, and showed similar activity against Gram-positive cocci. PMID- 10788843 TI - Clinical evaluation of the efficacy and tolerability of ritonavir in combination with two reverse transcriptase inhibitors. AB - A retrospective study of 76 patients was carried out using ritonavir in an antiretroviral regimen combined with two reverse transcriptase inhibitors to treat outpatients from July, 1996, to April, 1998, with the objective of evaluating clinical efficacy and tolerability. Seventy-six percent of the patients had been diagnosed with AIDS, an average number of CD(4) cells = 233.7 cells/mm(3) and viral load = 144, 084 RNA copies/mm(3). The majority of patients (76.3% )were antiretroviral treatment-experienced, 21.4% having taken protease inhibitors. A positive clinical response was found in 86.7% (including an average weight gain of 4.4 kg in 58.5% ), an average CD(4) count increase of 169.5 cells/mm(3) in 83.3% and an average viral load decrease of approximately 2.28 log in 75% of patients. A high percentage of adverse effects (76.3% ) was detected, with most slight or moderate, but they significantly impacted adherence to treatment as 31.6% stopped taking the drug as a result. We conclude that this antiretroviral regimen has good clinical efficacy, but relatively poor tolerability. PMID- 10788844 TI - Evaluation in an animal model and in vitro of the combination clavulanic acid and cephalosporins against beta-lactamase producing and nonproducing Staphylococcus aureus strains. AB - Beta-lactamase enzymes are the most common cause of bacterial resistance to Beta lactam antibiotics. They hydrolyze the amide bound in the Beta-lactam ring and produce acidic derivatives that have no antibacterial properties. The aim of this study was to evaluate a combination of clavulanic acid with cephalosporins against Beta-lactamase-producing and nonproducing strains of Staphylococcus aureus using in vitro tests and a rat animal model. In vitro tests (MIC) of the drug combination were done using standard methods. In an animal model, rats were submitted to surgical implantation of polyurethane sponges in their backs to induce granulomatous tissue. After seven days, the animals received cephalexin, cephalexin with clavulanic acid, ceftriaxone, ceftriaxone with clavulanic acid or clavulanic acid alone. One hour after the drug administration, granulomatous tissue was removed and placed in Petri dishes previously inoculated with 10(8) cfu of producing or non-producing Beta-lactamase Staphylococcus aureus. After 24h at 37 degrees C, the inhibition zones formed by granulomatous tissue was measured and scored for statistical analysis. Both tests (ex vivo ?animal model? and in vitro) showed that the cephalexin was more active than ceftriaxone against non producing Beta-lactamase S.aureus (p<0.01). Against Beta-lactamase producing S.aureus, ceftriaxone was more active than cephalexin, which was inactive. Combinations of clavulanic acid with cephalexin or ceftriaxone had similar antimicrobial activity against non-producing Beta-lactamase S.aureus compared to the cephalosporins used alone. When tested using Beta-lactamase producing strains, the combination of clavulanic acid with cephalosporins showed synergism. We conclude that the combination of cephalosporins with clavulanic acid could be useful in staphylococcal infections caused by Beta-lactamase producing strains. PMID- 10788845 TI - Conflicting messages regarding emerging microbial resistance, microbial sensitivity testing and control of antibiotic use in hospitals. PMID- 10788846 TI - Vertebroplasty: an opportunity to do something really good for patients. PMID- 10788847 TI - The efficacy of interconnected porous hydroxyapatite in achieving posterolateral lumbar fusion in sheep. AB - STUDY DESIGN: An animal study was performed to evaluate lumbar spinal fusion radiologically and mechanically. OBJECTIVES: To assess the efficacy of interconnected porous hydroxyapatite in achieving posterolateral lumbar arthrodesis in sheep. SUMMARY OF BACKGROUND DATA: Posterolateral spinal arthrodesis with autologous bone graft is the gold standard procedure for lumbar fusion. The procedure for harvesting bone from the iliac crest increases morbidity. Interconnected porous hydroxyapatite has been used effectively as an alternative to cancellous bone graft material in metaphyseal bone defects. Little is known about the efficacy of interconnected porous hydroxyapatite in achieving lumbar spinal fusion. METHODS: Four groups of seven sheep underwent bisegmental posterolateral lumbar fusion with instrumentation using different intertransverse graft material. In group 1, no graft material was used. In group 2, autologous bone was used. Group 3 had interconnected porous hydroxyapatite. Group 4 had an equip of interconnected porous hydroxyapatite and autologous bone. The animals were killed at 20 weeks after surgery. Radiographs and computed tomography images were obtained. The fusion masses were graded for bone resorption and trabecular connectivity on the computed tomography images. Mechanical testing of the specimens was performed, and the three-dimensional segmental motion was measured in flexion/extension, axial rotation, and lateral bending. RESULTS: The radiographic images were difficult to interpret because of the radiodense interconnected porous hydroxyapatite granules. According to mechanical stability criteria, the fusion rate for the different groups was as follows: 100% (14/14) for the autologous bone group, 72% (10/14) for the bone/interconnected porous hydroxyapatite group, 50% (7/14) for the pure interconnected porous hydroxyapatite group, and 15% (2/14) for the sham group. CONCLUSIONS: Spinal arthrodesis using interconnected porous hydroxyapatite alone or mixed with bone as graft material reduced segmental motion. It was not, however, as effective as autologous bone graft material in achieving spinal arthrodesis. The sheep model using autologous bone achieved a 100% fusion rate. Because the nonunion rate for a single level in humans may be as high as 40%, the fusion rate with bone/interconnected porous hydroxyapatite in humans may be lower than the 72% found in the sheep model. The little resorption of the radiodense interconnected porous hydroxyapatite granules made the radiologic evaluation of the fusion masses difficult. PMID- 10788848 TI - Biomechanical evaluation of a new bone cement for use in vertebroplasty. AB - STUDY DESIGN: Comparative ex vivobiomechanical study. OBJECTIVES: To determine the strength and stiffness of osteoporotic vertebral bodies subjected to compression fractures and subsequently stabilized via bipedicular injection of one of two bone cements: one is a commercially available polymethylmethacrylate (Simplex P) and one is a proprietary glass-ceramic-reinforced BisGMA/BisEMA/TEGDMA matrix composite that is being developed for use in vertebroplasty (Orthocomp). SUMMARY OF BACKGROUND DATA: Osteoporotic compression fractures present diagnostic and therapeutic challenges for the clinician. Vertebroplasty, a new technique for treating such fractures, stabilizes vertebral bodies by injection of cement. Little is known, however, about the biomechanics of this treatment. METHODS: Five vertebral bodies (L1-L5) from each of four fresh spines were harvested from female cadavers (age, 80 +/- 5 years), screened for bone density using DEXA (t = -3.4 to -6.4), disarticulated, and compressed in a materials testing machine to determine initial strength and stiffness. The fractures then were repaired using a transpedicular injection of either Orthocomp or Simplex P and recrushed. RESULTS: For both cement treatments, vertebral body strength after injection of cement was significantly greater than initial strength values. Vertebral bodies augmented with Orthocomp recovered their initial stiffness; however, vertebral bodies augmented with Simplex P were significantly less stiff than they were in their initial condition. CONCLUSIONS: Augmentation with Orthocomp results in similar or greater mechanical properties compared with Simplex P, but these biomechanical results have yet to be substantiated in clinical studies. PMID- 10788849 TI - The effects of cyclic loading on pull-out strength of sacral screw fixation: an in vitro biomechanical study. AB - STUDY DESIGN: The pull-out strength of sacral screw fixation after cyclic loading was tested using young human cadaveric specimens. OBJECTIVES: To evaluate the effects of fatigue loading on the pull-out strength of medial and lateral unicortical and bicortical sacral screws and to correlate the pull-out strength with sacral bone density and the screw insertion torque. SUMMARY OF BACKGROUND DATA: The immediate biomechanical effects of depth of penetration, screw orientation, and bone density on sacral screw fixation have been studied in aged cadaveric specimens. The effect of cyclic loading on the pull-out strength of sacral screw fixation is unknown, however, and data from young specimens is rare. METHODS: Eleven fresh specimens of human sacrum were used in this study. Bone mineral density at the vertebral body and the ala were determined by peripheral quantitative computed tomography. Seven-millimeter compact Cotrel-Dubousset sacral screws were inserted into the sacrum anteromedially and anterolaterally, both unicortically and bicortically, and the insertion torque for each screw was measured. Cyclic loading from 40 to 400 N was applied to each screw at a frequency of 2 Hz up to 20,000 cycles. Pull-out tests were conducted after completion of the fatigue tests. RESULTS: The average bone density was 0.38 +/- 0.08 g/mL at the S1 body and 0.24 +/- 0.05 g/mL at the S1 ala. The insertion torque and average pull-out force after cyclic loading were significantly higher for bicortical fixation than for unicortical fixation for a particular screw alignment. The pull-out strength and insertion torque of medially oriented fixation was always higher than that for lateral fixation, however, regardless of whether the insertion was unicortical or bicortical. The pull-out force of unicortical and bicortical medial screw fixations after cyclic loading showed significant linear correlations with both the insertion torque and the bone mineral density of the S1 body. CONCLUSIONS: In a young population, screw orientation (anterolateral or anteromedial) was more important in determining pull-out strength than screw depth (unicortical or bicortical) after fatigue loading, anteromedially directed screws being significantly stronger than laterallyplaced screws. Bone mineral density of the S1 body andinsertion torque were good preoperative and intraoperative indicators of screw pull-out strength. PMID- 10788850 TI - Experimental vertebroplasty using osteoconductive granular material. AB - STUDY DESIGN: Osteoporotic human cadaveric thoracic vertebral bodies and vertebral bodies from mature sheep were used as model systems to assess coral resorption and new bone formation after injection of coral granules. OBJECTIVE: To evaluate the use of natural coral exoskeleton, an osteoconductive material, for the filling of vertebral bodies. SUMMARY OF BACKGROUND DATA: Percutaneous injection of polymethylmetacrylate (PMMA) is often proposed for prophylactically stabilizing osteoporotic vertebral bodies at risk for fracture or augmentation of vertebral bodies that have already fractured. Recently, the possibility of using osteoconductive materials in granular formulation was assessed in pilot studies. METHODS: As a first step, the possibility of injecting coral granules percutaneously within osteoporotic human cadaveric thoracic vertebral bodies was assessed. As a second step, cavities were drilled into vertebral bodies of 10 mature ewes and were either left empty (control group) or filled with coral alone (CC) or coral supplemented with fibrin sealant (CC+FS). Quantitative evaluation of coral resorption and new bone formation was made 2 months and 4 months after implantation. RESULTS: The distribution of coral granules injected into human cadaveric thoracic vertebral bodies was homogenous as assayed radiographically. In the experimental animal model, osteogenesis was increased in cavities filled with coral in comparison with cavities left empty at both 2 months and 4 months (P < 0.005 and P < 0.02, respectively). Surprisingly, supplementation of coral with a fibrin sealant had no positive influence on osteogenesis (P < 0.0008 at 2 months; P < 0.002 at 4 months). In addition, it led to an increase in coral resorption by as soon as 2 months (P < 0.0008). CONCLUSION: These results demonstrate the osteoconductivity of coral in granular form for vertebral filling. Interestingly, interconnectivity between adjacent bone trabeculae and newly formed bone was restored; however, its mechanical significance remains to be determined. Further investigations are needed to evaluate the efficacy of coral in osteopenic animals and in relieving pain. PMID- 10788851 TI - Effect of implant design and endplate preparation on the compressive strength of interbody fusion constructs. AB - STUDY DESIGN: A human cadaveric study on the compressive strength of different lumbar interbody fusion implants and endplate preparation techniques was performed. OBJECTIVES: To assess the axial compressive strength of an implant with peripheral endplate contact as opposed to full surface contact, and to assess whether removal of the central bony endplate affects the axial compressive strength. SUMMARY OF BACKGROUND DATA: The compressive strength of interbody fusion constructs has been compared between implants and bone grafts. Neither implant design nor endplate preparation has been shown to affect strength. Removal of the central bony endplate for bone grafts was noted to improve graft incorporation but also to facilitate subsidence. METHODS: A total of 44 vertebrae were tested in four experimental groups by combining two interbody implants (full surface vs peripheral surface support) with two endplate preparation techniques (intact bony endplate vs removal of the central bony endplate). Specimens were tested to ultimate compressive failure using a 50 N/second ramped load. Yield strength and ultimate compressive strength were compared between groups using two factor analysis of covariance. A P value less than 0.05 was considered significant. Stepwise linear regressions assessed the predictive power of age, bone mineral content, and the implant's normalized endplate coverage on yield strength and ultimate compressive strength. RESULTS: Neither implant design nor endplate preparation technique affected yield strength or ultimate compressive strength. Age, bone mineral content, and the normalized endplate coverage were strong predictors of yield strength (P < 0. 0001; r2 = 0.459) and ultimate compressive strength (P < 0.0001; r2 = 0.510). CONCLUSIONS: An implant with only peripheral support resting on the apophyseal ring offers axial mechanical strength similar to that of an implant with full support. Neither supplementary struts nor a solid implant face has any additional mechanical advantage, but reduces graft-host contact area. Removal of the central bony endplate is recommended because it does not affect the compressive strength and promotes graft incorporation. PMID- 10788852 TI - Sagittal plane configuration of the sacrum in spondylolisthesis. AB - STUDY DESIGN: A radiographic study of the sagittal sacral deformity in spondylolisthesis. OBJECTIVES: To characterize and classify the pathoanatomy of sagittal sacral deformation in spondylolisthesis. SUMMARY OF BACKGROUND DATA: Spondylolisthesis has been extensively described and reviewed in the literature. Deformity of the entire sacrum in spondylolisthesis potentially could affect the natural history, treatment options, and outcome. The sagittal contour of the entire human sacrum has never been quantitatively studied in spondylolisthesis. METHODS: A literature search was performed and data was gathered retrospectively on patients with spondylolisthesis at the authors' institution. Cases of degenerative spondylolisthesis were excluded. Specifically those patients with L5 S1 spondylolisthesis were studied. The authors studied standing lateral radiographs and performed statistical analysis to understand morphologic relations. RESULTS: A broad range of global sacral kyphosis (37-188 degrees ) exists in spondylolisthesis. Increasing sacral kyphosis is significantly associated with increasing percent slip, sacral horizontal angle, Neuman's classification, lumbar lordosis, and lumbar index. A simple classification of the spectrum of sacral deformity in the sagittal plane is presented. CONCLUSION: The entire sacrum in spondylolisthesis can develop a significant kyphotic deformity in the sagittal plane, and this is associated with other abnormalities found in the lumbosacral spine. Sacral deformity is a significant factor in the assessment of the sagittal contour of the patient with L5-S1 spondylolisthesis. PMID- 10788853 TI - Progressive rotational dislocation in kyphoscoliotic deformities: presentation and treatment. AB - STUDY DESIGN: Progressive rotational dislocation of the spine has been described as the most serious evolutive risk of kyphoscoliosis. A retrospective chart review was conducted on 11 patients with this deformity. OBJECTIVES: To delineate the clinical and radiologic characteristics of this entity to facilitate early diagnosis and treatment. The outcome after treatment was analyzed to point out the rationale for appropriate treatment. METHODS: The characteristic radiologic feature was a short sharp angled kyphosis (average 112) at the junction of two lordoscoliotic curvatures. The etiology of the spinal deformity was neurofibromatosis in four patients and various dysplastic conditions in seven patients. Two patients had congenital vertebral defects. Structural weakness of the bone was therefore a basic feature. Neurologic impairment was identified in three patients (one complete, two incomplete). Four patients had a nonunion after a previous attempt at spinal fusion: two after a combined anterior and posterior fusion with an anterior approach from the convexity and two after a posterior fusion alone. All patients underwent complete circumferential stabilization through anterior strut-grafting and posterior fusion. An anterior approach from the concavity was performed systematically with tibial strut grafts inserted in a palisade fashion. Preoperative correction of the deformity was performed by progressive controlled elongation in a Stagnara elongation cast. Cotrel-Dubousset instrumentation was used in two patients, Harrington instrumentation was used in two patients, and cast immobilization alone was used in seven patients. RESULTS: The average follow-up period was 5 years 5 months. All but one patient achieved successful spinal fusion. Loss of correction at the latest follow-up evaluation was less than 3 degrees in nine patients. The two patients with incomplete neurologic deficits were improved, but the patient with the complete deficit remained unchanged after surgery. CONCLUSIONS: Awareness of the possibility of a progressive rotational dislocation in dystrophic forms of kyphoscoliosis should allow for an early diagnosis and stabilization. The percentage of patients having a neurologic deficit in this series was significantly less important than in the initial report. Early anterior strut grafting from the concavity of the scoliotic curvature and posterior fusion is recommended. PMID- 10788854 TI - The iliolumbar ligament: three-dimensional volume imaging and computer reformatting by magnetic resonance: a technical note. AB - STUDY DESIGN: An assessment of magnetic resonance imaging techniques of the iliolumbar ligament. OBJECTIVES: To identify a technique to better image the iliolumbar ligament. SUMMARY OF BACKGROUND DATA: The iliolumbar ligament plays an important role in providing lumbosacral stability. Two-dimensional magnetic resonance imaging of the lumbar spine only provides a fragmented representation of the ligament. METHODS: A two-part study was performed. In the first part, three cadaver pelvi were dissected, and the individual bands of the iliolumbar ligament were identified. Computer reformatting of magnetic resonance three dimensional volume images then were performed, correlating the structural characteristics of the iliolumbar ligament to its magnetic resonance image. In the second part of the study, the lumbosacral region of three groups of patients were evaluated. Group I was studied with routine magnetic resonance imaging techniques of the intervertebral disc regions. Group II was studied with routine contiguous axial magnetic resonance imaging of the lumbosacral spine. Group III was studied with computer reformatting of three-dimensional volume images of the lumbosacral spine. RESULTS: Accurate imaging of the iliolumbar ligament of cadaver specimens was achieved with three-dimensional volume imaging and computer reformatting. Routine imaging of the intervertebral disc region as well as contiguous axial imaging of the spine depicted only limited segments of the iliolumbar ligament. Three-dimensional volume imaging and computer reformatting allowed precise imaging of the iliolumbar ligament in all patients, demonstrating the ligament orientation as well as length, width, and depth. CONCLUSION: Only images of the iliolumbar ligament obtained through computer reformatting of three dimensional volume averaging from L3 to the sacral ala correlated with the ligament's structural characteristics. PMID- 10788855 TI - Post-traumatic findings of the spine after earlier vertebral fracture in young patients: clinical and MRI study. AB - STUDY DESIGN: A study comparing magnetic resonance imaging findings of degenerative changes in intervertebral discs in young patients with previous wedge-shaped compression fracture and age-matched and sex-matched control subjects. OBJECTIVES: To find out the role of fractures in disc degeneration and to assess the clinical outcome of the patients. SUMMARY OF BACKGROUND DATA: Several experimental studies have postulated that trauma is one of the major reasons for disc degeneration. Wedge compression fractures in vertebrae of children have been considered insignificant, but this has not been verified in the literature. METHODS: Fourteen patients 8.8 to 20.8 years of age (mean, 15.5 years) with a history of wedge-shaped vertebral compression fracture at least 1 year previously (mean, 3.8 years) and asymptomatic healthy control subjects were studied by thoracolumbar spine magnetic resonance imaging. The patients also underwent a clinical examination. RESULTS: Eight (57%) of the 14 patients had disc degeneration, and seven of them had it at the trauma level. Of these 7 subjects, 6 also had endplate damage at this level. The association between endplate damage and adjacent intervertebral disc degeneration was significant (P < 0.01). Only 2 of the patients were symptomatic. In the control group, only 1 subject had disc degeneration with endplate changes and disc herniation. CONCLUSIONS: The patients had more disc degeneration than did those in the control group. Endplate injury was strongly associated with disc degeneration. No correlation between previous vertebral fracture and back pain was seen in this study. PMID- 10788856 TI - The factors associated with neck pain and its related disability in the Saskatchewan population. AB - STUDY DESIGN: Population-based, cross-sectional mailed survey. OBJECTIVE: To identify factors associated with neck pain and its related disability in Saskatchewan adults. SUMMARY OF BACKGROUND INFORMATION: Little is known about the etiology of neck pain and its related disability. Previous cross-sectional population-based studies have suggested that neck pain may be associated with age, female gender, lower socioeconomic status, physically demanding work, and other comorbidities. METHODS: The Saskatchewan Health and Back Pain Survey was mailed to 2184 randomly selected Saskatchewan adults 20 to 69 years of age. Fifty five percent of the study population participated. The survey collected demographic, socioeconomic, and health-related information. Neck pain and its related disability was classified into four categories using the Chronic Pain Questionnaire: no neck pain (Grade 0), low intensity/low disability neck pain (Grade I), high intensity/low disability neck pain (Grade II), and high disability neck pain (Grades III-IV). Polytomous logistic regression was used to identify associations between demographic, socioeconomic, and health-related variables and various grades of neck pain severity. RESULTS: Of the 1131 respondents, 54% had experienced neck pain at some point in the 6 months before the survey, and almost 5% were highly disabled by neck pain. The prevalence of Grade I neck pain was lower in individuals with low education attainment, but higher for those reporting headaches, low back pain, better general health, and a history of neck injury resulting from a motor vehicle collision, some of whom may have received compensation for their injury. Grade II neck pain was strongly associated with headache, low back pain, and a history of neck injury during a motor vehicle collision and weakly associated with digestive disorders and current cigarette smoking. Grades III-IV neck pain was strongly associated with low back pain, headaches, cardiovascular disorders, digestive disorders, and a history of neck injury during a motor vehicle collision. CONCLUSION: This study suggests that important associations exist between comorbidities, a past history of neck injury resulting from a motor vehicle collision, and graded neck pain. Importantly, individuals who are significantly disabled by neck pain also have comorbidities that have a moderate or severe impact on their health, suggesting that chronic disorders tend to cluster in some individuals. PMID- 10788857 TI - Spinal posture during stooped walking under vertical space constraints. AB - STUDY DESIGN: Simultaneous spine kinematic variables in sewage workers were quantified using a two-dimensional video-based gait analysis system. OBJECTIVES: To identify patterns of spinal posture in a population of sewage workers pushing waste matter through tunnels while walking stooped under various height constraints. SUMMARY OF BACKGROUND DATA: Working with stooped postures is one of several occupational risk factors that have been associated with spinal disorders. However, the specific changes in spinal posture during stooped walking under various height constraints have not been documented. METHODS: A video-based gait analysis system was used to measure spinal posture in 22 sewage workers. Angles of the cervical, thoracic, and lumbar spine in the sagittal plane were assessed during walking with five levels of height constraint, from upright walking to stooped walking under a headroom restriction of 105 cm. Correlations among gait parameters, demographic data, and clinical results were determined. Linear regression analysis was performed to examine which variables have the largest impact on the posture resulting from a given vertical height constraint when age and body height are held constant. RESULTS: Mean angular values changed significantly with increasing headroom restrictions, with increases in cervical and thoracic extension as well as lumbar flexion. The cervical and thoracic angles were best correlated with height constraint, followed by lumbar angle and stride length. The inverse relation between cervical and thoracic angle during upright walking increased with increasing vertical space constraints, whereas the relation between the thoracic and lumbar angles decreased. Subjects with decreased abdominal muscle strength adopted a significantly more kyphotic thoracic posture when walking under headroom constraints than subjects with normal abdominal muscle strength. CONCLUSIONS: Combined walking and pushing under vertical space constraints was associated not only with the expected increased flexion of the lumbar spine, but also with greater extension (i.e., reduced kyphosis) of the thoracic spine. PMID- 10788858 TI - The role of anticipation and fear of pain in the persistence of avoidance behavior in patients with chronic low back pain. AB - STUDY DESIGN: A correlative design using stepwise regression analysis. OBJECTIVE: To explore the variation in spinal isometric strength that can be accounted for by anticipation of pain, sensory perception of pain, functional disability belief, and the fear-avoidance belief in chronic low back pain patients. SUMMARY OF THE BACKGROUND DATA: Several biobehavioral factors contribute to the persistence of pain behavior in chronic patients. Recent studies suggest a need to explore the relation between reduced physical performance and the sensory and cognitive perception of pain. METHODS: Sixty-three patients with chronic low back pain 20 to 56 years of age participated in this study. Visual Analogs Scales, the Fear-Avoidance Belief questionnaire, and the Disability Belief questionnaire were used to measure the sensory and cognitive dimensions of pain. Spinal isometric strength was measured by the Medx lumbar extension machine. RESULTS: Analysis of variance and the stepwise regression analysis demonstrated that anticipation of pain and the fear-avoidance belief about physical activity significantly predicted variation in the spinal isometric strength deficit P < 0. 001. True pain experienced during the testing and answers to the Disability Belief questionnaire were not related. CONCLUSION: The results of this study strongly support the hypothesis that spinal physical capacity in chronicity is not explained solely by the sensory perception of pain. The anticipation of pain and the fear-avoidance belief about physical activities were the strongest predictors of the variation in physical performance. PMID- 10788859 TI - Cost-effectiveness of fusion with and without instrumentation for patients with degenerative spondylolisthesis and spinal stenosis. AB - STUDY DESIGN: A cost-effectiveness study was performed from the societal perspective. OBJECTIVE: To evaluate the costs and benefits of laminectomy alone and laminectomy with concomitant lumbar fusion for patients with degenerative lumbar spondylolisthesis and spinal stenosis. SUMMARY OF BACKGROUND DATA: Costs, probabilities, and utilities were estimated from the literature. Short-term risks considered were perioperative complications, the probability of the fusion healing, and the probability that surgery will relieve symptoms. Long-term risks considered were recurrence of symptoms and reoperation. METHODS: The 10-year costs, quality-adjusted life years, and incremental cost-effectiveness ratios (reported as dollars per quality-adjusted year of life gained) were calculated using a Markov model. Sensitivity analysis was performed on all variables using clinically plausible ranges. RESULTS: Laminectomy with noninstrumented fusion costs $56,500 per quality-adjusted year of life versuslaminectomy without fusion. The cost-effectiveness of laminectomy with noninstrumented fusion was most sensitive to the increase in quality-of-life associated with relief of severe stenosis symptoms. The cost-effectiveness ratio of instrumented fusion compared with noninstrumented fusion was $3,112,800 per quality-adjusted year of life. However, if the proportion of patients experiencing symptom relief after instrumented fusion was 90% as compared with 80% for patients with noninstrumented fusion, then the cost-effectiveness ratio of instrumented fusion compared with noninstrumented fusion would be $82,400 per quality-adjusted year of life. CONCLUSIONS: The cost-effectiveness of laminectomy with noninstrumented fusion compares favorably with other surgical interventions, although it depends greatly on the true effectiveness of these surgeries to alleviatesymptoms and on how patients value the quality-of-life effect of relieving severe stenosis symptoms. Instrumented fusion was very expensive compared with the incremental gain in health outcome. Better data on the effectiveness of these alternative procedures are needed. PMID- 10788860 TI - The test of Lasegue: systematic review of the accuracy in diagnosing herniated discs. AB - STUDY DESIGN: A systematic review of the literature including statistical meta analysis. OBJECTIVES: To evaluate published methods of the test of Lasegue or straight leg raising test and the cross straight leg raising test by using a recently developed criteria list and to summarize and explore reasons for variation in diagnostic accuracy. SUMMARY OF BACKGROUND DATA: Little evidence exists on the diagnostic accuracy of the widely used straight leg raising test and the cross straight leg raising test in diagnosing herniated discs in patients with low back pain. METHODS: MEDLINE and EMBASE searches up to 1997 showed 17 diagnostic publications evaluating the straight leg raising test with surgery as reference standard. Quality of methods was assessed with a specific checklist. Eleven studies were selected for statistical pooling. Sources of variation and heterogeneity were studied by meta-regression of the diagnostic odds ratio. RESULTS: All studies were surgical case-series at nonprimary care level. Verification-bias was obvious in one study. Pooled sensitivity for straight leg raising test was 0. 91 (95% CI 0.82-0.94), pooled specificity 0.26 (95% CI 0.16 0.38). Pooled diagnostic odds ratio was 3.74 (95% CI 1.2-11.4). Discriminative power was lower in recent studies, in studies with only inclusion of primary hernias, and with blind assessment of both the index-test (straight leg raising test) and the reference (surgery). For the cross straight leg raising test pooled sensitivity was 0.29 (95% CI 0.24-0.34), pooled specificity was 0.88 (95% CI 0.86 0.90), and the pooled diagnostic odds ratio 4.39 (95% CI 0.74-25.9). CONCLUSIONS: The diagnostic accuracy of the straight leg raising test is limited by its low specificity. Discriminative power decreased with a more valid design, a more homogenous case-mix, and year of publication. Although the studies may reflect everyday clinical practice, they do not enable a valid evaluation of the diagnostic accuracy of both tests. Diagnostic research should evaluate the validity of the complete diagnostic process and study the evidence of the added value of the different tests used. [Key words: sensitivity, specificity, diagnosis, meta-analysis, test of Lasegue, straight leg raising test] PMID- 10788861 TI - A review of psychological risk factors in back and neck pain. AB - STUDY DESIGN: The literature on psychological factors in neck and back pain was systematically searched and reviewed. OBJECTIVES: To summarize current knowledge concerning the role of psychological variables in the etiology and development of neck and back pain. SUMMARY OF BACKGROUND DATA: Recent conceptions of spinal pain, especially chronic back pain, have highlighted the role of psychological factors. Numerous studies subsequently have examined the effects of various psychological factors in neck and back pain. There is a need to review this material to ascertain what conclusions may be drawn. METHODS: Medical and psychological databases and cross-referencing were used to locate 913 potentially relevant articles. A table of 37 studies was constructed, consisting only of studies with prospective designs to ensure quality. Each study was reviewed for the population studied, the psychological predictor variables, and the outcome. RESULTS: The available literature indicated a clear link between psychological variables and neck and back pain. The prospective studies indicated that psychological variables were related to the onset of pain, and to acute, subacute, and chronic pain. Stress, distress, or anxiety as well as mood and emotions, cognitive functioning, and pain behavior all were found to be significant factors. Personality factors produced mixed results. Although the level of evidence was low, abuse also was found to be a potentially significant factor. CONCLUSIONS: Psychological factors play a significant role not only in chronic pain, but also in the etiology of acute pain, particularly in the transition to chronic problems. Specific types of psychological variables emerge and may be important in distinct developmental time frames, also implying that assessment and intervention need to reflect these variables. Still, psychological factors account for only a portion of the variance, thereby highlighting the multidimensional view. Because the methodologic quality of the studies varied considerably, future research should focus on improving quality and addressing new questions such as the mechanism, the developmental time factor, and the relevance that these risk factors have for intervention. PMID- 10788862 TI - Successful short-segment instrumentation and fusion for thoracolumbar spine fractures: a consecutive 41/2-year series. AB - STUDY DESIGN: A retrospective review of all the surgically managed spinal fractures at the University of Missouri Medical Center during the 41/2-year period from January 1989 to July 1993 was performed. Of the 51 surgically managed patients, 46 were instrumented by short-segment technique (attachment of one level above the fracture to one level below the fracture). The other 5 patients in this consecutive series had multiple trauma. These patients were included in the review because this was a consecutive series. However, they were grouped separately because they were instrumented by long-segment technique because of their multiple organ system injuries. OBJECTIVES: The choice of the anterior or posterior approach for short-segment instrumentation was based on the Load Sharing Classification published in a 1994 issue of Spine. The purpose of this review was to demonstrate that grading comminution by use of the Load-Sharing Classification for approach selection and the choice of patients with isolated fractures who are cooperative with spinal bracing for 4 months provide the keys to successful short-segment treatment of isolated spinal fractures. SUMMARY OF BACKGROUND DATA: The current literature implies that the use of pedicle screws for short-segment instrumentation of spinal fracture is dangerous and inappropriate because of the high screw fracture rate. METHODS: Charts, operative notes, preoperative and postoperative radiographs, computed tomography scans, and follow-up records of all patients were reviewed carefully from the time of surgery until final follow-up assessment. The Load-Sharing Classification had been used prospectively for all patients before their surgery to determine the approach for short-segment instrumentation. Denis' Pain Scale and Work Scales were obtained during follow-up evaluation for all patients. RESULTS: All patients were observed over 40 months except for 1 patient who died of unrelated causes after 35 months. The mean follow-up period was 66 months (51/2 years). No patient was lost to follow-up evaluation. Prospective application of the Load-Sharing Classification to the patients' injury and restriction of the short-segment approach to cooperative patients with isolated spinal fractures (excluding multisystem trauma patients) allowed 45 of 46 patients instrumented by the short segment technique to proceed to successful healing in virtual anatomic alignment. CONCLUSIONS: The Load-Sharing Classification is a straightforward way to describe the amount of bony comminution in a spinal fracture. When applied to patients with isolated spine fractures who are cooperative with 3 to 4 months of spinal bracing, it can help the surgeon select short-segment pedicle-screw-based fixation using the posterior approach for less comminuted injuries and the anterior approach for those more comminuted. The choice of which fracture dislocations should be strut grafted anteriorly and which need only posterior short-segment pedicle-screw-based instrumentation also can be made using the Load Sharing Classification. PMID- 10788863 TI - Vertebral artery occlusion after acute cervical spine trauma. AB - STUDY DESIGN: A retrospective study of vertebral artery injury diagnosed during the last 6 years in our institution. OBJECTIVES: To determine the clinical and radiologic features of vertebral artery injury. SUMMARY OF BACKGROUND DATA: Extracranial occlusion of the vertebral artery associated with cervical spine fracture is uncommon and can cause serious and even fatal neurologic deficit due to back lifting and cerebellar infarction. Magnetic resonance imaging and magnetic resonance angiography are extremely helpful in the examination of acute injuries of the cervical spine. METHODS: Magnetic resonance imaging and magnetic resonance angiography were performed at the time of injury. RESULTS: The authors reviewed six patients with cervical spine fractures who were diagnosed with a unilateral occlusion of the vertebral artery by means of magnetic resonance imaging/magnetic resonance angiography. One patient had signs of vertebrobasilar insufficiency and another with complete cord lesion had cerebellar and back lifting infarctions. Surgical anterior spinal fusion was performed in five patients, and one was treated by traction and orthosis. At the time of discharge, five patients had no vertebrobasilar symptoms, and the patient who experienced vertebrobasilar territory infarctions showed no progression of the neurologic damage. CONCLUSIONS: Vertebral artery injury should be suspected in cervical trauma patients with facet joint dislocation or transverse foramen fracture. Magnetic resonance imaging/magnetic resonance angiography is a helpful test to rule out vascular injury. Vertebral artery injury affects the extracranial segment at the same level as the cervical fracture. This is a retrospective review that did not permit drawing conclusions about the effects of early surgical stabilization in the treatment of cervical spine injuries with associated vertebral artery injury; however, surgical stabilization may avoid propagation and embolization of the clot located at the site of the lesion. PMID- 10788864 TI - Os odontoideum with cerebellar infarction: a case report. AB - STUDY DESIGN: A case report. OBJECTIVES: To report the case of a child with os odontoideum associated with cerebellar infarction and to discuss the correlation between atlantoaxial instability with os odontoideum and vertebrobasilar artery insufficiency. SUMMARY OF BACKGROUND DATA: Knowledge of the influence of atlantoaxial instability on vertebrobasilar artery insufficiency remains limited despite the publication of several reports. METHODS: A 5-year-old boy with ataxic gait disturbance was hospitalized in the pediatric ward. Magnetic resonance imaging revealed multiple cerebellar infarctions, and cerebral angiogram showed occlusions of several branches of the basilar artery and a winding of the left vertebral artery. Stress lateral radiographs of the cervical spine showed atlantoaxial instability with os odontoideum. Posterior C1-C2 transarticular screw fixation with iliac bone graft was applied to obtain firm stability and fusion. RESULTS: There was no damage to the vertebral arteries or spinal nerves in the perioperative period. Solid union of the grafted bone and rigid stability of the atlantoaxial joint were seen on lateral flexion-extension radiographs 1 year after the operation. There has been no sign of recurrent arterial insufficiency, and the patient has been free from cerebellar dysfunction to date. CONCLUSIONS: Atlantoaxial instability may cause insufficiency of the vertebral artery as well as spinal cord injury. More attention should be paid to the possible relation between atlantoaxial instability and vertebrobasilar artery insufficiency. PMID- 10788865 TI - A post-traumatic ligamentum flavum progressive hematoma: a case report. AB - STUDY DESIGN: A case report. OBJECTIVES: To understand a rare case of ligamentum flavum progressive hematoma. SUMMARY OF BACKGROUND DATA: Previously there were only two reports about ligamentum flavum hematoma. METHODS: A patient was surgically treated for ligamentum flavum hematoma causing progressive L5 radiculopathy. Clinical and neuroradiologic features were reported, and the literature was reviewed. RESULTS: The etiology of this case could not be defined except by minor back injury. In spite of conservative therapy, the symptoms were progressive for 7 months. Magnetic resonance imaging demonstrated the epidural mass lesion at L4-L5 that was continuous with the ligamentum flavum. The mass was hypointense in T1-weighted images and central hyperintense and marginal hypointense in T2-weighted images. The margin was well enhanced by Gd-DTPA administration. After removal of the mass lesion, the patient's symptoms completely resolved. Before surgery, accurate diagnosis was difficult even based on magnetic resonance imaging and was achieved after histologic examinations. CONCLUSIONS: Surgery could be a choice of the treatment modality to resolve symptoms in ligamentum flavum hematoma. PMID- 10788867 TI - Imaging corner PMID- 10788868 TI - Imagery PMID- 10788866 TI - Eikenella corrodens vertebral osteomyelitis secondary to direct inoculation: a case report. AB - STUDY DESIGN: A case report. OBJECTIVE: To present a unique method of inoculation with an unusual bacterial organism, Eikenella corrodens, which led to vertebral osteomyelitis and to heighten awareness of different bacterial organisms that may cause orthopedic infections. SUMMARY OF BACKGROUND DATA: E. corrodens has been known to cause orthopedic infections for more than 20 years. The usual mechanism is from "Fight bite" injuries to the hand or metacarpal phalangeal joint. Eikenella osteomyelitis of the vertebral body is extremely rare. METHODS: A 65 year-old man from Indonesia developed slowly progressive vertebral collapse and worsening neck pain. History suggested a previous pharyngeal injury while eating fish. Work up for malignancy was negative, and imaging studies were suggestive of osteomyelitis. An open biopsy and culture as well as fusion were performed for treatment of the osteomyelitis. RESULTS: After a complete work up was undertaken, cultures grew out E. corrodens as the causative organism of this patient's osteomyelitis. Computed tomography scan and history were consistent with inoculation of the paravertebral space with E. corrodens secondary to a fish bone transgressing the patient's pharynx. The patient was treated with surgery and the appropriate antibiotics and healed without any significant sequela. CONCLUSION: Vertebral osteomyelitis due to E. corrodens is extremely rare. Direct inoculation as a cause of vertebral osteomyelitis with Eikenella has never been reported. If diagnosed properly with appropriate cultures and antibiotic sensitivities, Eikenella osteomyelitis can be treated successfully according to standard orthopedic practices including debridement, fusion, and appropriate antibiotics. Awareness of unusual bacteria as potential causative organisms of osteomyelitis is imperative for appropriate treatment. PMID- 10788869 TI - Changing a suprapubic catheter--1. PMID- 10788870 TI - Nurses gave warning that the staffing crisis was a disaster waiting to happen. PMID- 10788871 TI - Eyes wide open. PMID- 10788872 TI - NHS stalwarts starved of recognition. PMID- 10788873 TI - Pressure gauge. PMID- 10788874 TI - Delivery reward. PMID- 10788875 TI - Money talks. PMID- 10788876 TI - The new EU directive for formula milk companies. PMID- 10788877 TI - Conference season found both government and opposition fired up over health. PMID- 10788878 TI - Occupational health works. PMID- 10788879 TI - A painful legacy. PMID- 10788880 TI - Inside the double helix. PMID- 10788881 TI - Invisible link. PMID- 10788882 TI - Women should not be able to opt for a caesarean. PMID- 10788883 TI - Nursing strategy. Leading from the front. PMID- 10788884 TI - Intensive care. The final reckoning. PMID- 10788885 TI - Understanding how nursing affects our psychological and general health could help to improve conditions in the NHS. PMID- 10788886 TI - Menorrhagia: the treatment options. PMID- 10788887 TI - Plague: the flea-borne horror story. PMID- 10788888 TI - Carers tap into the information highway. PMID- 10788890 TI - We're independent, are you? PMID- 10788889 TI - How does it feel to be somebody's lifeline? PMID- 10788891 TI - Continence. Looking for a new direction. PMID- 10788893 TI - Continence. Services for a new century. PMID- 10788892 TI - Belfast lessons in continence care. PMID- 10788894 TI - Continence. Geared up for guidance. PMID- 10788895 TI - Continence. Direct to the public. PMID- 10788896 TI - Continence. Bangladeshi venture. PMID- 10788897 TI - Continence. International trends. PMID- 10788898 TI - Reassessing the care of patients at the end of life: an opportunity to reunify our profession. PMID- 10788899 TI - Writing for publication as an advanced practice nurse. AB - Advanced practice nurses (APN) develop and test nursing interventions, design initiatives in their organizations, and propose new ways of delivering care to patients and families; however, they frequently do not communicate these innovations to others. This article presents strategies for incorporating professional writing into the role of APN. PMID- 10788900 TI - The personal and organizational costs of blowing the whistle. PMID- 10788901 TI - Learning outcomes of integrative preceptorships. AB - The purpose of this study was to identify and compare nursing faculty, preceptor, and student perceptions of ideal and actual learning outcomes of integrative preceptorships for senior generic baccalaureate nursing students. The sample included senior generic baccalaureate nursing students involved in preceptorships, their registered nurse preceptors, and the faculty responsible for the experiences. Findings revealed areas of significant differences between the preceptors' and the other two groups' perceptions of ideal and actual learning outcomes. When faculty, preceptor, and student perceptions of ideal learning outcomes were compared with their perceptions of what was actually achieved through the experience, significant differences emerged for all three groups. PMID- 10788902 TI - Stress and the caregiver. AB - What do we do with a stress level that is high in many areas of our life? How do nurses cope without becoming ill themselves? We look at the literature for documented causes of the stress that seems to pervade the workplace and for strategies that will help us to function as healthy human beings. PMID- 10788903 TI - Ethical and human rights concerns of Connecticut nurses: survey and implications for the profession. AB - A survey of a random sample of Connecticut registered nurses' experiences with issues of ethics and human rights found a strong focus on protecting patient rights and dignity, adequacy of staffing patterns, informed consent, and respecting advance directives. Ethics committees were commonly available but were infrequently used. Participants reported high levels of ethics education but also desired more such education. The character of the issues is consistent with other studies and largely reflects conflicts over which nurses do not have independent control. Consideration is given to issues of nurses' moral agency in health care organizations. PMID- 10788905 TI - The Inspector General and JCAHO. PMID- 10788904 TI - Quality of care-risk adjustment outcomes model: testing the effects of a community-based educational self-management program for children with asthma. AB - This article describes the design of an outcomes model incorporating adjustment for patients' risks for various outcomes of care. A community-based educational self-management program for children with asthma is proposed for testing the model. Measures of model components: clinical factors; nonclinical factors; psychological, cognitive, and psychosocial functioning; and patient and client satisfaction are described. Testing this model has implications for guiding the development of individualized culturally sensitive nursing interventions for children with asthma and their parents. PMID- 10788906 TI - [A life with kidney failure, transplantation and dialysis. "Then there is no more fear"]. PMID- 10788907 TI - [Preparation for kidney transplantation]. PMID- 10788908 TI - [Use of a nursing model in the operating room--a study]. PMID- 10788909 TI - [Age and society]. PMID- 10788910 TI - [Nursing aspects of chronic patient care]. PMID- 10788912 TI - [The Hartberg social models]. PMID- 10788911 TI - [Patients' instructions: an effective form of patients' consent?]. PMID- 10788913 TI - Home healthcare: what's in the cards? PMID- 10788914 TI - The ethical considerations of outcome research. AB - Escalating costs have led to the increasing interest in outcome research in all areas of industry. In healthcare, providers, payors, and consumers are making every effort to improve the quality of patient care while managing costs. Because the overriding issue of cost is driving much of outcome research, it is increasingly important to recognize the fragile balance that exists between cost and quality. In addition, clinicians face a number of other ethical and legal concerns such as confidentiality, informed consent, and the needs of the underserved. This column describes outcome research, along with ethical principles that influence this research. PMID- 10788915 TI - Teaching a blind patient colostomy irrigation. PMID- 10788916 TI - International Wound Care Ambassadorship. Health and wound care in The Netherlands. PMID- 10788917 TI - HIV/AIDS: impact on healing. AB - Human immunodeficiency virus and acquired immune deficiency syndrome (HIV/AIDS) are leading chronic illnesses in many major cities worldwide. Frequently, people with HIV infection require surgery or develop chronic wounds. This paper summarizes the impact of HIV infection on body organs and systems and the effect of antiretroviral therapy as a basis for potential complications with wound care and healing. The authors also present research on wound healing in HIV-positive people undergoing operative procedures. Besides the physical effect of HIV infection on the person, clinicians must also realize psychosocial and economic effects of the disease when considering wound care. This paper also addresses care considerations for patients with HIV/AIDS in the inpatient, outpatient, and home care settings. PMID- 10788918 TI - Enhanced healing and cost-effectiveness of low-pressure oxygen therapy in healing necrotic wounds: a feasibility study of technology transfer. AB - Recent advances in topical hyperbaric oxygen technology identified the use of low pressure topical hyperbaric oxygen therapy in enhancing wound healing. This study prospectively examined the feasibility of technology transfer from university to Health Maintenance Organization personnel, using topical hyperbaric oxygen therapy to heal necrotic wounds. Fifteen patients with 24 gangrenous and/or necrotic wounds that did not improve or worsened after at least 6 weeks of standard wound care were treated with topical hyperbaric oxygen therapy by trained HMO personnel. Four patients underwent digital amputation for osteomyelitis and/or gangrene followed by topical hyperbaric oxygen therapy. Assessment parameters included wound healing and cost of wound care before and after topical hyperbaric oxygen therapy. Six of the six Level 2 wounds healed within 2 to 4 weeks, nine of the ten Level 3 wounds healed within 4 to 10 weeks, and seven of the eight Level 4 wounds healed within 4 to 12 weeks. The ulcers improved by a mean of 0.829 cm2 per day. T test (SSPS 7.5) showed significant improvement per day after topical hyperbaric oxygen therapy, t = 5.217, df = 24, P < 0.0001 (95% CI = 1.13-0.49). Wound healing with topical hyperbaric oxygen therapy was associated with decreased costs. The results of this support the feasibility of transfer of new wound healing technology from research to HMO personnel. PMID- 10788919 TI - Therapeutic and financial outcomes using a pulsating low-air-loss surface for patients surviving severe posterior burns. AB - Patients with posterior burns require extensive stays in the intensive care unit for recovery. The authors hypothesized that pulsating low-air-loss therapy would decrease the intensive care unit length of stay for burn patients, resulting in a potentially significant reduction in charges to payors. Eighty-one posterior burn patients enrolled in the primary study were randomly assigned to a pulsating low air-loss surface (study group) or a nonpulsating low-air-loss surface (control group). The 54 survivors in this analysis (the secondary study) were well matched for age, pre-existing conditions, and total body surface area burned. Average intensive care unit length of stay was less for the study patients compared with the control patients--40 days versus 64 days (P < .05). Control patients used specialty surfaces for 49 days and study patients used them for 38 days. Based on a daily intensive care unit charge of $1,000 and the average daily specialty surface rental charge, the study patients averaged potential charges of under $44,000 in comparison to more than $67,000 for control patients. These data suggest that treatment of posterior burns with pulsating low-air-loss therapy may be of great clinical and financial benefit, decreasing the intensive care unit length of stay and potentially contributing to reduced charges to payors. PMID- 10788921 TI - Gene therapy for healthcare providers. PMID- 10788920 TI - Joining the French in wound care: it's a small world after all. PMID- 10788922 TI - Examining caregiver rights. AB - Lay caregivers increasingly are accepting responsibility for the care of loved ones in the home health setting. These caregivers are often not prepared emotionally or physically for such responsibility. The rights of caregivers are examined herein as they relate to the obligations of clinicians to ease the caregivers transition to their new and sometimes unexpected role. PMID- 10788923 TI - Lower extremity wound: what's your assessment? PMID- 10788924 TI - Photographic assessment of the appearance of chronic pressure and leg ulcers. AB - The purpose of this paper was to examine the validity and reliability of using photographs of wounds to accurately assess wound status. The results of assessing wound appearance using wound photographs was compared to results obtained from a bedside assessment using the Pressure Sore Status Tool (PSST). The photographic wound assessment tool (PWAT) used in this comparison represents a modified version of the PSST and includes the six domains that can be determined from wound photographs. The PWAT was used on photographs of both chronic pressure ulcers (n = 56) and leg ulcers due to vascular insufficiency (n = 81). The photographic tool has excellent intrarater (ICC = 0.96) and interrater (ICC = 0.73) reliability and good concurrent validity (r = 0.70) compared with a full bedside assessment PSST. The PWAT has also shown to be sensitive to change in wound appearance of healing ulcers, but not nonhealing ulcers. These results would suggest that in the event that a full bedside assessment is not possible, wound photographs may be used to accurately assess wound appearance of both chronic pressure ulcers located on the trunk and vascular ulcers of the lower extremity. Establishing a valid and reliable assessment of wound healing using photographic images is of great relevance to the advancing fields of computer image analysis and telemedicine. PMID- 10788926 TI - Pulsed lavage: promoting comfort and healing in home care. AB - The purpose of this study was to determine if hydrodebridement with pulsed lavage facilitates the removal of necrotic tissue, promotes healing, and increases comfort in the homebound patient. Home healthcare provides cost-effective care in the setting that is most conducive to healing. People want to be at home, yet many illnesses require services that cannot be obtained at home. Wound care sometimes falls into this category. Pulsed lavage has expanded treatment options for the homebound patient. Hydrodebridement with pulsed lavage is site specific, avoids cross contamination, and is less expensive than whirlpool therapy. It also may facilitate the removal of necrotic tissue and promote the formation of healthy granulation tissue. The authors performed a retrospective audit to gather data on 28 patients who received pulsed lavage treatments at home. A descriptive analysis of five variables that affect healing was undertaken including mobility/activity, nutritional status, cardiovascular/respiratory status, continence, and sensory perception. All of the clients in the sample achieved a clean, warm, moist wound bed, free of signs and symptoms of infection, absence of necrotic tissue, and the presence of granulation tissue to meet the definition of "ready for healing" as presented in the literature. The majority of clients experienced no pain. Although comorbid conditions required rehospitalization for 35.7% of the sample, the conditions did not interfere with healing. Hydrodebridement with pulsed lavage is a viable nontraumatic, noninvasive, site specific treatment alternative for patients receiving care in the home. PMID- 10788925 TI - Prevalence and types of wounds among children receiving care in the home. AB - The purpose of this study was to determine the number of children with wounds receiving home care, determine the types of wounds among these children, and identify wound care products used in the treatment of children. This study was a multi-site, descriptive, cross-sectional, collaborative, study involving 13 home healthcare agencies and a university. Nurses (n = 281) were systematically selected to collect data for patients receiving home care visits during a 1-week period. The nurses recorded data on 77 children who ranged in age from less than 1 year to 18 years (mean = 3.1 years, SD = 4.72). The children included 47 males and 30 females, most of whom were African-American (n = 59). Among the 77 children, 16.9% had wounds. Children with wounds were significantly older and had more reasons for the home healthcare visit than children without wounds. The presence of a wound was not significantly related to the length of the visit, gender, or race. The most common wound was a surgical incision. The wound care treatments used most were tap water and gauze. In conclusion, wound care is frequently part of the care for a child in the home. Therefore, nurses need to be aware of wound assessment and wound care protocols that match a child's growth and development and family needs. PMID- 10788927 TI - ["To every wound a human being is attached"]. PMID- 10788928 TI - [Health insurance agencies have 9.5 billion marks on the high edge]. PMID- 10788929 TI - [Drug therapy of type 2 diabetes: insulin sensitizers improve results]. PMID- 10788930 TI - [Prevention of thromboembolism: compression dressing yields best results]. PMID- 10788931 TI - [Fecal incontinence: nursing care has a key position]. PMID- 10788932 TI - [Thermometry in nursing: new studies lead to interesting results]. PMID- 10788933 TI - [No more fear of complaints: complaints considered as chance]. PMID- 10788934 TI - [Quality assurance through documentation of care: nursing care under the magnifying glass]. PMID- 10788935 TI - [Hindering competence: what are serious nursing errors?]. PMID- 10788936 TI - [The justifying consent: every treatment is legally a potential injury]. PMID- 10788937 TI - [Leave spent with institutionalized mental patients: closeness and trust generate or assure security]. PMID- 10788938 TI - [Health care system in the Land of Thousand Lakes: in Finland nursing takes over medical care]. PMID- 10788939 TI - [Nuns and nursing care in the middle of the 20th century]. AB - The aim of this qualitative study was to investigate the connection between the life of nine nuns in the Augustian order, simultaneously working as nurses in the middle of the 20th century. To get an extensive idea about this period, the nuns, now 60 to 84 years old, were asked about their life at that time in narrative interviews. They had all worked between 1930 and 1960 as nurses. Through content analysis according to Mayring (1997) the authors generated the core category "We have always been there". This category shows that nuns constantly had to be present for patients, nurses, medication and the head of the order, that they were held responsible for all care and that they had to be available for the order and the hospital all day and night. The head of the order determined where they had to work. These irregular working hours and the fact that they frequently took over management positions prematurely led young nuns beyond their physical and psychological limits. Their religion should serve as "source of effort" for their work as nurses. PMID- 10788940 TI - [Bertha Pappenheim (1859-1936) and her role in nursing care]. AB - In 1954 the German federal postal service issued a special stamp displaying Bertha Pappenheim (1859-1936) in the series "Helfer der Menschheit" (Helpers for mankind) together with Kathe Wollwitz, Lorenz Werthmann und Johann Friedrich Oberlin. Despite being honoured elsewhere, too, her engagement has not been taken note of by any biographers dealing with the history of nursing. This article which conveys a survey of her life and work therefore concentrates primarily on the importance Bertha Pappenheim gave nursing. PMID- 10788941 TI - [Nursing in the profession of the "sub-surgeons" between 1750 and 1850]. AB - Between 1750 and 1850 surgical training was professionalized as a consequence of newly established technical colleges for surgeons in Germany and Austria led by doctors rather than craftsmen. At first, anatomy, debridement and obstetrics was taught, then a technical knowledge of medical treatment in general was instructed at specific medical/surgical schools. Depending on their years of training, graduates then worked as country doctors or as surgical assistants ("Subchirurgen", second class wound doctors). Nursing care was also on the curriculum; therefore some students worked temporarily as nurses. After graduating, many supervised nursing care in hospitals. This profession died out due to developments in the medical profession of the mid-19th century. However, they never belonged to the proletarian class of orderlies. PMID- 10788942 TI - [Perception of symptom distress in lung cancer patients: I. Agreement between patients and their caregiving relatives]. AB - This paper employs a comparative descriptive design to compare primary family caregivers' assessment of lung cancer patients' symptom distress with patients' own perceptions of symptom distress in the home setting. The second part describes the results of the qualitative component of this research. A convenience sample of 37 patient-family caregiver dyads completed the McCorkle and Young Symptom Distress Scale (SDS). Family caregivers' global scores were moderately correlated with patients' global scores (r = 0.71; P < 0.001). No significant differences in ratings were found for 10 of the 13 symptoms assessed. Therefore, when the patient is unable to provide a self-report of symptom distress, health-care professionals may seriously consider family caregivers' assessments of patients' symptom distress to be reasonable estimates for at least 10 of the 13 symptoms on the SDS. PMID- 10788943 TI - [Mediation of transcultural nursing care in clinical context: a tightrope walk]. AB - Our wish for suggestions for a line of action regarding our contact with migrants in the clinical context has led us to an increased acceptance of "Transcultural Nursing" by Madeleine Leininger. However, an uncritical adoption of this theory without theoretical and historical background information produces in practice a stereotyped image of migrants, and a cultural rating of social and individual aspects instead of mutual understanding. Hence, from the present-day perspective, Leininger's model is not suitable to establish transcultural nursing care in practice. A theory not encouraging the analysis of socio-cultural backgrounds of one's own but the sight "from without" of the so-called alien--a theory that does not make interaction its main object but culture, cannot serve as a basis for transcultural nursing care. Up-to-date transcultural nursing care does not need the introduction of a specific nursing concept for migrants but an extension of nursing by a socio-cultural and migration-specific dimension. However, nursing care does not seem to accept this challenge, what with it being far easier to add the "Leininger Culture Care Theory" to the existing curricula as an additional model. Thus nursing loses the opportunity of grappling with the adoption of certain theories and models of other disciplines. Even though Leininger is herself a nurse, she has developed her model in her capacity as a cultural anthropologist using the background of anthropological theories. But, the supplying of transcultural nursing care demands also from cultural anthropologists an analysis of the concrete context of nursing care and its approaches. Only by bearing this in mind, cultural anthropology can counter effectively the danger of being marginalized as a fringe discipline, and of being reduced to the level of providing cooking recipes. PMID- 10788944 TI - [Nursing science and related sciences. Questions about centralization of nursing science and interdisciplinary relations]. AB - The thesis of this paper is that nursing science draws a lot of its innovative potential from the knowledge that has been gathered in related disciplines. This knowledge is specified and reformulated with regard to nursing practice. The thesis is developed on two levels: first under aspects of methodology and the philosophy of science, then under aspects of the adoption of interdisciplinary knowledge. The integration of various related sciences that takes place in nursing science will finally be demonstrated in an example. This example shows which qualifications the practitioners need to deal with patients who suffer from a stroke. PMID- 10788945 TI - [Comparison of the effectiveness and cost of treatment with humid environment as compared to traditional cure. Clinical trial on primary care patients with venous leg ulcers and pressure ulcers]. AB - INTRODUCTION: The discovery of moist environment dressings as alternatives to the traditional treatments based on exposing wounds to air, opened new expectations for the care and treatment of chronic wounds. Over the years, these expectations have led to the availability of new moist environment dressings which have made it possible to improve the care provided to patients suffering this kind of wounds, as well as providing important reasons to weigh in terms of cost-benefit effectiveness at the time of selecting which type of treatment should be employed. The lack of comparative analysis among traditional treatments and moist environment treatments for chronic wounds among patients receiving primary health care led the authors to perform an analysis comparing these aforementioned options of treatment on patients suffering venous leg ulcers or pressure ulcers. PATIENTS, MATERIALS AND METHODS: The authors designed a Randomized Clinical Trial involving patients receiving ambulatory care in order to compare the effectiveness and cost-benefit of traditional versus moist environment dressing during the treatment of patients suffering stage II or III pressure ulcers or venous leg ulcers. In this trial, variables related to effectiveness of both treatments, as well as their costs were analyzed. MAIN RESULTS: 70 wounds were included in this Randomized Clinical Trial, 41 were venous leg ulcers of which 21 received a moist environment treatment while 20 received traditional cure, the other 29 wounds were pressure ulcers of which 15 received moist environment dressings treatment and 14 received traditional dressings. No statistically significant differences were found among the defining variables for these lesions in either group under treatment. In the venous leg ulcer study group, the authors conclusions were an average of 18.13 days, 16.33 treatment sessions and a cost of 10,616 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with traditional treatment compared to an average of 18.22 days, 4.54 treatment sessions and a cost of 2409 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with moist environment dressings. In the pressure ulcers study group, the authors conclusions were an average of 12.18 days, 12.1 treatment sessions and a cost of 15,490 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with traditional treatment compared to an average of 7.12 days, 1.86 treatment sessions and a cost of 2610 pesetas to heal one square centimeter of the initial surface area of a wound on patients treated with moist environment dressings. COMMENTS: The results of this randomized clinical trial demosntrated that the moist environment treatment group was more effective and had a better cost-benefit ratio than the traditional treatment group in the treatment of pressure ulcers and venous leg ulcers on patients cared for by nursing personnel in primary health care centers all of which agrees with publications consulted by authors. PMID- 10788946 TI - [Health education during menopause. A long-range evaluation]. AB - After annually evaluating the repercussions that a health program during menopause has on women, The authors carry out a study to determine if the results obtained last over a much longer time period. This cross-sectional study was carried out in 1998 on a random sample taken from all the women who participated in this program during the years 1995 and 1996. Furthermore, the authors made comparisons between both sample year groups. The study results demonstrate that once women have acquired the knowledge necessary to serve as a foundation to practice healthy habits, they integrate these habits into their life style on a permanent basis. Part of this paper was presented orally in the Andalucian Conference on Nursing in Gynecology which took place in November 1998 in Seville. PMID- 10788947 TI - [Wounds sutured by nurses at the Jaen City Hospital]. PMID- 10788948 TI - A personal experience. PMID- 10788949 TI - Leading mergers and acquisitions in the health care market--tips for nurse managers and nurse executives. PMID- 10788950 TI - Capital investment decision making. PMID- 10788952 TI - Surviving mergers and acquisitions PMID- 10788951 TI - Mergers and acquisitions: a socially acceptable mechanism for redeploying productive capacity. PMID- 10788953 TI - Managing in turbulent times: issues and challenges in health care mergers and acquisitions. AB - The environment of the health care system in the present and foreseeable future has been described as a revolution whose impetus comes from Wall Street. The new system of health care is characterized by mergers, acquisitions, and joint ventures. For-profit conversions and mergers of religious and secular organizations were almost unheard of before the last decade. The challenges facing nursing leadership in these turbulent times are (1) dealing with the human dynamics associated with creating new organizational cultures, (2) shifting focus away from event-driven cost avoidance and protecting institutional assets, and (3) shifting focus toward stewardship of community resources and nursing practice beyond institutional boundaries. PMID- 10788955 TI - 'Uncharted waters'--consolidation of two academic psychiatric services: a case study. AB - Surviving during times of mergers and acquisitions is difficult but possible. Following is a case study of how one service managed not only to survive, but to thrive, during consolidation. PMID- 10788954 TI - Mergers and acquisitions in professional organizations: a complex adaptive systems approach. AB - Nurse managers face unique challenges as they cope with mergers and acquisitions among health care organizations. These challenges can be better understood if it is recognized that health care institutions are professional organizations and that the transformations required are extremely difficult. These difficulties are caused, in part, by the institutionalized nature of professional organizations, and this nature is explicated. Professional organizations are stubborn. They are repositories of expertise and values that are societal in origin and difficult to change. When professional organizations are understood as complex adaptive systems, complexity theory offers insight that provide strategies for managing mergers and acquisitions that may not be apparent when more traditional conceptualizations of professional organizations are used. Specific managerial techniques consistent with both the institutionalized characteristics and the complex adaptive systems characteristics of professional organizations are offered to nurse managers. PMID- 10788956 TI - An interview with Brenda Ernst: surviving a complex merger in an academic medical center. Interview by Elizabeth A. Buck. PMID- 10788957 TI - Creativity on the edge of chaos. AB - The health care environment is chaotic, with many threats and problems. Despite nursing's resulting discomfort, being able to see the opportunities and the potential within such complexity is freeing. The author illustrates the use of creative thinking in formulating deliberate responses to the unpredictability of chaos. With such innovative actions, the essential nature of professional nursing can be reaffirmed. PMID- 10788959 TI - Surviving mergers and acquisitions--as you sow, so shall you reap.... PMID- 10788958 TI - Caring for yourself during times of organizational change. AB - Mergers, acquisitions, and other organizational changes in the health care system pose many challenges and stresses for nursing leaders. Although considerable effort is expended helping staff adjust to change, nurse managers and executives often neglect their own vulnerability to occupational stress. The personal and organizational costs of managerial burnout are high, sometimes resulting in physical illness, mental exhaustion, and decreased productivity. This article emphasizes the importance of prevention through the recognition of potential or actual signs of organizational stress. Handy tips are provided on effective ways to cope with the stress of organizational change. PMID- 10788960 TI - Calcium supplements. PMID- 10788961 TI - Bexarotene (Targretin) for cutaneous T-cell lymphoma. PMID- 10788962 TI - Intravenous equipment--the ongoing development of the syringe. PMID- 10788963 TI - Dental trauma associated with anaesthesia. AB - Damage to teeth is the most common complaint against anaesthetists. A dental history and oral examination are important before anaesthesia. Pre-existing dental pathology or the presence of prostheses makes damage more likely but sound teeth may be affected. The maxillary central incisors are most at risk. Certain diseases and drugs should alert anaesthetists to increased likelihood of dental pathology. The flange of the Macintosh blade appears responsible for much damage and alternative equipment or techniques of endotracheal intubation should be considered, particularly when risk factors are present. Manoeuvres to protect teeth must not impact adversely on airway management. Custom mouthguards can be useful. A management plan can help control losses if damage does occur. Patients should be warned about the possibility of dental trauma. PMID- 10788964 TI - Intubation conditions and postoperative myalgia in outpatient dental surgery: a comparison of succinylcholine with mivacurium. AB - Ninety-four patients undergoing elective outpatient third molar extraction were recruited into a double-blind, randomized, prospective trial comparing mivacurium (group M) with succinylcholine (Group S) for conditions for endotracheal intubation and the occurrence of postoperative myalgia. Anaesthesia was induced with fentanyl 1 microgram.kg-1 and propofol 2.5 micrograms.kg-1 in all patients. Group S patients were given gallamine 20 mg while group M patients were given mivacurium 0.2 mg.kg-1. Manual ventilation was commenced and anaesthesia maintained with nitrous oxide 70% and isoflurane 1 to 2% in oxygen. After two minutes, group S patients were given succinylcholine 1.5 mg.kg-1 and group M patients 0.9% saline. Nasotracheal intubation was performed 30 seconds later. Intubating conditions in group M were significantly better than those in group S (P < 0.001). The incidence of postoperative myalgia was 9.5% in group M and 26% in group S but this was not statistically significant (P = 0.09). We propose that mivacurium is a suitable neuromuscular blocker to use for endotracheal intubation in outpatient dental surgery. PMID- 10788965 TI - A comparison of carbon dioxide monitoring and oxygenation between facemask and divided nasal cannula. AB - The divided nasal cannula is a device recently released in Australia that couples oxygen delivery and end-tidal carbon dioxide (PETCO2) monitoring. This study compares the accuracy of PETCO2 measurements by the divided nasal cannula and those measured by a modified facemask (as currently used in this institution), with arterial partial pressure of carbon dioxide (PaCO2). In this crossover study, 30 patients who had arterial lines as part of their routine monitoring were given oxygen via nasal cannula and facemask preoperatively. The PETCO2 was measured with each device and a simultaneous PaCO2 and PaO2 measured after equilibration. The results demonstrate a significant difference between the PETCO2 as measured by each technique. The divided nasal cannula more accurately reflects PaCO2 (mean arterial to end expired gradient of 5 mmHg) and provides a more representative trace when compared to a traditional facemask system. Both methods provided adequate oxygenation. PMID- 10788966 TI - The dose-effect relationship for morphine and vomiting after day-stay tonsillectomy in children. AB - A dose-response curve for intravenous morphine and vomiting was investigated in children having day-stay tonsillectomy. A retrospective chart review was performed for the 164 children fulfilling the inclusion criteria. Morphine (mean 0.09 mg/kg SD 0.05) was used in 108 children in the perioperative period and a further 56 children were given no opioid. Fifty-five of these 164 children vomited and 20 children required an overnight stay in hospital because of vomiting. The probability of vomiting or overnight stay in hospital was related to morphine dose (by logistic regression). The overall probability of vomiting after morphine 0.1 mg/kg was 50% and the probability of admission for vomiting with this dose was 10%. Pharmacodynamic parameter estimates for postoperative vomiting were P0 (the baseline probability of vomiting, with no opioid) 0.115, Pmax (the maximal probability of vomiting due to morphine) 0.997, ED50 (morphine dose that induces an effect equivalent to 50% of the logit Pmax) 0.18 mg/kg. Parameter estimates for overnight stay because of vomiting after morphine administration were P0 0.038, Pmax 0.999, ED50 0.369 mg/kg. Satisfactory postoperative analgesia in children has been reported with morphine 0.05 to 0.15 mg/kg. Doses above 0.1 mg/kg are associated with a greater than 50% incidence of vomiting. Our data suggests that lower doses of morphine are associated with a decreased incidence of emesis after tonsillectomy in children. PMID- 10788967 TI - Pulmonary toxicity of inhaled aerosolized prostacyclin therapy--an observational study. AB - Large white/landrace piglets (mass 11 to 21 kg) were exposed to aerosolized alkaline glycine diluent (n = 2) or inhaled aerosolized prostacyclin (n = 2) for five to eight hours. Pigs receiving these aerosols developed mild acute sterile tracheitis, involving the superficial layers of the trachea, shown histologically and ultrastructurally. Pigs receiving the diluent aerosol also showed mild inflammatory changes in the bronchioles. These findings suggest caution with the use of high volumes of aerosolized alkaline glycine diluent during inhaled aerosolized prostacyclin therapy. PMID- 10788968 TI - Immunoassays in the diagnosis of anaphylaxis to neuromuscular blocking drugs: the value of morphine for the detection of IgE antibodies in allergic subjects. AB - Radioimmunoassays (RIAs) for IgE antibodies to specific neuromuscular blocking drugs (NMBDs) are an important tool in the diagnosis of anaphylaxis during anaesthesia although they are performed in only a few laboratories throughout the world. NMBDs bind to antibodies by their substituted ammonium ions. We measured serum IgE antibodies to morphine and specific NMBDs in 347 patients with suspected anaphylaxis using blood specimens sent for mast cell tryptase assays. Morphine, which has a single substituted ammonium group, avidly binds in vitro to antibodies that react with NMBDs. The morphine RIA proved to be both a more sensitive and efficient test for the detection of IgE antibodies to NMBDs than the specific NMBD RIAs. We have adopted the morphine RIA in our laboratory in preference to the specific RIAs and predict that use of this single assay will become widespread for the in vitro diagnosis of allergic sensitivities to NMBDs. PMID- 10788969 TI - An analysis of excess mortality not predicted to occur by APACHE III in an Australian level III intensive care unit. AB - The APACHE III derived standardized mortality ratio has been suggested as a statistic to measure intensive care unit (ICU) effectiveness. From 1991 data collected on 519 consecutive admissions to the Royal Adelaide Hospital ICU a standardized mortality ratio of 1.25 was calculated. Of the 174 deaths only 95 had a prediction of death greater than 0.5. As part of a quality assurance study we undertook a retrospective case note audit to try to identify factors that were associated with the low mortality prediction (< 0.5) in hospital deaths. Firstly we analysed the patient population that died to determine the factors that were different between patients who had a mortality prediction of greater than 0.5 versus those who had a mortality prediction of less than 0.5. Next we analysed the patient population with a mortality prediction of less than 0.5 and compared actual survivors with patients who died in hospital. Amongst low mortality prediction patients admitted to the Royal Adelaide Hospital ICU we identified age, a history of acute myocardial infarction, presentation to ICU after a cardiac arrest or with an elevated creatinine and the development of acute renal failure and septicaemia during the ICU admission as being associated with in hospital mortality. We also documented that late hospital deaths on the ward after ICU discharge occurred more frequently with low predicted hospital mortality ICU patients. Factors other than the APACHE III score may be associated with hospital deaths of ICU patients. PMID- 10788970 TI - Hours of work and fatigue-related error: a survey of New Zealand anaesthetists. AB - A nationwide survey (70% response) documented anaesthetists' hours of work, their perceptions about safety limits and their recollection of fatigue-related errors in clinical practice. In the preceding six months, 71% of trainees and 58% of specialists had exceeded their self-defined safety limits for continuous anaesthesia administration. For 50% of trainees and 27% of specialists, their average working week exceeded their own limits for maintaining patient safety, and for 63% of trainees and 40% of specialists, it exceeded their limits for maintaining their personal well-being. Fatigue-related errors were reported by 86% of respondents, with 32% recalling errors in the preceding six months. Specialists were more likely to report a fatigue-related error if they had exceeded their own safety limits for continuous anaesthesia administration, or for weekly work hours. Current measures are not preventing anaesthetists from working hours that they consider to be unsafe for patients or harmful to their own well-being. PMID- 10788971 TI - The European Donor Hospital Education Programme (EDHEP): enhancing communication skills with bereaved relatives. AB - Intensive care medical and nursing staff self-rate their communication skills as improved following attendance at the European Donor Hospital Education Programme (EDHEP) workshop. A prospective study was conducted to determine what impact EDHEP has on communication skills. Doctor-nurse pairs from 10 experimental and 10 control Intensive Care Units undertook two standardized simulated relative encounters (Breaking Bad News and Donation Request) at three measurement points (pre, post and follow-up). Nurses showed no change in communication skills. Experimental group doctors showed significant improvement in breaking bad news and requesting donation; most of these improvements were not maintained. Control group doctors showed some improvement in breaking bad news, indicating that participating in measurement by itself initiates some transient change in communication skills. Attendance at EDHEP does lead to significant improvement in some, but not all, communication skills essential in breaking bad news and requesting donation. Further research is necessary to determine what factors additional to EDHEP will contribute to enduring change in these particular skills. PMID- 10788972 TI - The use of end-tidal carbon dioxide monitoring to confirm intratracheal cannula placement prior to percutaneous dilatational tracheostomy. AB - We tested the utility of intratracheal carbon dioxide monitoring (IT-CO2) in 10 patients undergoing percutaneous dilatational tracheostomy (PDT). We have found IT-CO2 monitoring reliable in confirming the correct position of the tracheal cannula prior to tracheal dilatation using the Portex technique. PMID- 10788973 TI - Submental intubation: an alternative to short-term tracheostomy. AB - Surgeons, anaesthetists and intensivists need to be reminded of an alternative to tracheostomy in the management of patients with severe maxillofacial injuries. The technique and indications for submental intubation are described. PMID- 10788975 TI - Severe hyponatraemia secondary to desmopressin therapy in von Willebrand's disease. AB - A 42-year-old female with von Willebrand's disease was managed with desmopressin and tranexamic acid to aid haemostasis following a vaginal hysterectomy. Severe acute hyponatraemia (134 to 108 mmol/l) developed over two days, culminating in a generalized tonic-clonic seizure and cerebral oedema. Fluid restriction, cessation of desmopressin and hypertonic saline administration led to a full recovery. Desmopressin is known to reduce free water elimination and produce hyponatraemia, but its extent and rate of development in this patient was surprising. Close monitoring of serum sodium and fluid balance is recommended in these patients. PMID- 10788974 TI - Anaesthetic management of a patient with low tracheal obstruction requiring placement of a T-Y stent. AB - A 60-year-old man with a history of oesophagectomy for carcinoma presented with worsening dysphagia, dyspnoea and tachypnoea secondary to oesophageal stricture and tumour invasion of the trachea causing a "ball-valve" obstruction. The patient required placement of a T-Y tracheobronchial stent to relieve the airway obstruction and was successfully managed using sevoflurane for gaseous induction followed by maintenance with propofol total intravenous anaesthesia. PMID- 10788976 TI - A case for steroids in acute lung injury associated with the retinoic acid syndrome. AB - A 62-year-old woman with acute promyelocytic leukaemia was treated with all-trans retinoic acid. On day 2 she suffered with dyspnoea and general fatigue. Marked hypoxia suggested the occurrence of retinoic acid syndrome. She underwent endotracheal intubation and mechanical ventilation with the administration of dexamethasone. Her symptoms promptly abated. She was subsequently treated with conventional chemotherapy and achieved complete remission. PMID- 10788977 TI - Epileptiform activity in the EEGs of two nonepileptic children under sevoflurane anaesthesia. AB - Two case reports of nonepileptic children are presented, who developed paroxysmal EEG potentials in routinely performed EEG recordings during inhalation of sevoflurane, 7 and 8% by volume respectively. Taking into account several reports from the literature about epileptiform potentials or convulsive movements under similar conditions, it seems to be important to investigate carefully the circumstances under which these phenomena appear as well as possible clinical consequences. PMID- 10788978 TI - Video-assisted thoracoscopic surgical pleurodesis for persistent spontaneous pneumothorax in late pregnancy. AB - A case of persistent spontaneous pneumothorax in the third trimester of pregnancy managed by video-assisted thoracoscopic surgical pleurodesis is presented. Anaesthetic and perioperative considerations are discussed. PMID- 10788979 TI - Anaesthetic management of a parturient with a colloid cyst of the third ventricle. AB - A colloid cyst in the third ventricle near the foramen of Monroe can obstruct cerebrospinal fluid (CSF) flow from the lateral ventricles. Any change in the CSF pressure on either side of the cyst can lead to displacement and thus precipitate acute hydrocephalus. Management of the confinement of a patient with a colloid cyst must therefore aim to minimize changes in CSF pressure. We describe our management of a patient with a small colloid cyst who was permitted to labour with the assistance of patient-controlled epidural analgesia. The available alternatives are discussed. PMID- 10788980 TI - Prevention of tube occlusion caused by biting: oral bite block versus oropharyngeal airway. PMID- 10788981 TI - What is the risk of aspirin withdrawal in the surgical patient? PMID- 10788982 TI - The Portex loss-of-resistance syringe. Check before using. PMID- 10788983 TI - Drager Julian workstation. PMID- 10788984 TI - Adult day case tonsillectomy. PMID- 10788985 TI - [Perioperative medicine: the significance of arteriosclerotic diseases]. PMID- 10788986 TI - [Prevention of perioperative myocardial ischemia--an update]. AB - Perioperative cardiac morbidity and mortality are a major health care challenge with important individual as well as economic aspects. Up to 30% of all perioperative complications and up to 50% of all postoperative deaths are related to cardiac causes. Perioperative myocardial ischemia, which occurs in more than 40% of patients with or at risk for coronary artery disease and undergoing noncardiac surgery, represents a dynamic predictor of postoperative cardiac complications. Long-duration myocardial ischemia and ischemic episodes associated with myocardial cell damage are particularly of prognostic relevance. In patients suffering from this type of ischemia, the incidence of adverse cardiac outcome is increased up to 20-fold. Reducing the incidence of perioperative myocardial ischemia is associated with a decrease in adverse cardiac outcome. Important issues related to perioperative myocardial ischemia are hematocrit level, body temperature, and hemodynamic variables. In contrast, the choice of anesthetic agents and techniques appears to be less important. Perioperative administration of anti-ischemic drugs in patients at risk, however, leads to a further decrease in the incidence of myocardial ischemia and to an improvement in patient outcome. Recent studies suggest that alpha 2-agonists and particularly beta-adrenoreceptor blocking agents are effective anti-ischemic drugs in the perioperative setting. Perioperative administration of beta-adrenoreceptor blocking agents in coronary risk patients undergoing noncardiac surgery is associated with a reduced rate of postoperative cardiac complications and an improvement in long-term outcome. This is particularly relevant in high risk patients with preoperative stress-induced ischemic episodes. In clinical practice, therefore, chronically administered anti ischemic drugs should also be administered on the day of surgery and during the postoperative period. In untreated patients with or at risk for coronary artery disease and who have to undergo urgent surgical procedures without the opportunity of preoperative anti-ischemic intervention, perioperative administration of beta-adrenoreceptor blocking agents is mandatory. PMID- 10788988 TI - [The effect of different volume expanders on neutrophil granulocyte function in vitro]. AB - INTRODUCTION: The influence of kolloids on the immune system is not well documented. In this study we investigated the effects of gelatine, hydroxyethylstarch (HES), human albumine, and dextrane on neutrophil function and receptor expression by flow cytometry. METHODS: Whole blood of healthy volunteers was incubated for 30 minutes with either gelatine, HES (6% and 10%), dextrane 40 and 60, or human albumin 20%. Phagocytic capacity was determined by uptake of fluorescein-isothiocyanate labeled bacteria, the conversion of dihydrorhodamine 123 into fluorescent rhodamine 123 was used for oxidative burst measurements. Expression of complement receptors CD 11b and CD35 was investigated using fluorescein-isothiocyanate labeled antibodies. RESULTS: Incubation with gelatine significantly increased expression of complement receptors and oxidative burst. Dextranes and HES had no influence on neutrophil function. Human albumin reduced the oxidative burst, whereas CD 35 expression was increased. CONCLUSION: The physiological significance of these changes in a range of 10% has to be clarified in further investigations. PMID- 10788987 TI - [Anesthetic complications. The incidence of severe anesthetic complications in patients and families with progressive muscular dystrophy of the Duchenne and Becker types]. AB - During the last 30 years a great number of case reports presented severe anaesthetic complications with sudden cardiac arrest in patients with muscular dystrophies, mostly unsuspected at the time of the event. As succinylcholine was involved in the majority of the intractable incidents with lethal outcome the Food and Drug Administration (FDA) of the United States recommended a warning of the administration of succinylcholine in young children and adolescents in 1992 and an extensive international discussion on the routine use of succinylcholine in paediatric anaesthesia. Epidemiological studies on this issue are rare. We projected an inquiry about the incidence rate and type of severe anaesthetic complications in an utmost large number of patients and families with Duchenne (DMD) and Becker type (BMD) muscular dystrophy. METHODS: With the approval of the ethic committee of the university Witten/Herdecke and informed consent of the participants we investigated all patients and families who were diagnosed, controlled and treated for DMD or BMD as inpatients or outpatients in a "Muscle Centre" since 1983. The questionnaire asked for the number of patients per family, classification of the disease DMD or BMD, number and date of anaesthetics in the patients and eventual complications, anaesthetics and eventual complications in the parents, siblings and relatives and the occurrence of malignant hyperthermia (MH) in the family or relatives. Statistical assessments were done by Fisher's exact test for stratified 2 x 2 tables and Zelen's test for homogeneity of odds ratios. RESULTS: 200 out of 224 questionnaires could be evaluated. The diagnosis was confirmed by molecular genetic and immunohistochemical investigations. In 147 families it turned out to be DMD, in 53 families BMD. The 212 male and 9 female patients in the 200 families were given 444 anaesthetics. Sudden cardiac arrest occurred in 6 patients, all successfully resuscitated. Nine less severe incidents consisted of fever, symptoms of rhabdomyolysis (CK-elevation, dark coloured urine, hyperkalemia) and masseter spasm. The statistical assessment revealed that the occurrence of an event was highly dependent whether the diagnosis of muscular dystrophy was established or not (p < 0.0001, Fisher's exact test). All six cardiac arrests occurred in the 45 families with undiagnosed disease and no event happened in the 134 families with already known DMD/BMD. There was evidence that the number of anaesthetics without prior establishment of the diagnosis decreased after 1992 (p = 0.004, Fisher's exact test). CONCLUSIONS: Our results demonstrate that severe incidents and cardiac arrests occurred only in young children with undiagnosed DMD or BMD who received inhalational agents and succinylcholine. A cardiac arrest in 6 out of 200 families was found much more frequently than in the normal paediatric population (about 1:1000 to 1:3000). The decrease of events after 1992 (warning of the FDA) and disappearance of sudden cardiac arrests in our group of patients might be due to the world wide discussion on routine use of succinylcholine in children or the much earlier establishment of the diagnosis in our population. An early diagnosis of DMD and BMD and the avoidance of the triggering agents succinylcholine and volatile anaesthetics can reduce the risk of severe anaesthetic complications. PMID- 10788989 TI - [Ciaglia blue rhino: a modified technique for percutaneous dilatation tracheostomy. Technique and early clinical results]. AB - OBJECTIVE: Elective tracheostomy has become an established treatment modality in modern intensive care medicine, and the number of percutaneously performed tracheostomies is steadily increasing. The Ciaglia Blue Rhino (CBR) represents another percutaneous technique in which the tracheostoma is formed by one-step dilation. Our study presents the technique itself and the early clinical experiences. METHODS: In 20 adult patients on long-term ventilation, CBR was done. After puncture of the trachea in typical manner, dilation of the tracheostoma was achieved in one single step by means of a curved dilator with a special hydrophilic coating. Then, the tracheostomy tube was inserted over a curved loading dilator. Practicability and safety were determined as well as gas exchange during the procedure by means of arterial blood gas samples. RESULTS: As a result of the dilator's hydrophilic coating, dilation of the tracheostoma was rapidly achieved within 152 +/- 22 s, virtually free from resistance of the trachea or the cervical tissues. Complications such as bleeding, aspiration or postoperative infection of the stoma were not noted in any of our patients. Fractures of isolated tracheal cartilage rings were seen in 5 patients, however, no therapeutic intervention was necessary. In terms of perioperative gas exchange, pre- and postoperative levels of FiO2, PaO2, PaCO2 and the oxygenation index (PaO2/FiO2) did not change significantly. CONCLUSIONS: Based on our early clinical experience, Ciaglia Blue Rhino represents a new method that may combine the typical advantages of each of the other techniques for percutaneous tracheostomy (i.e. Ciaglia, Griggs, Fantoni) in one single technique. This method is distinguished by a high level of safety and practicability. However, further comparative trials need to be done before a definitive judgement can be made. PMID- 10788990 TI - [Semi-invasive cardiac output measurement using a combined transesophageal ultrasound device. Early experiences]. AB - OBJECTIVE: Measurement of cardiac output (CO) with pulmonary artery catheter (PAC) is currently item of many discussions. We investigated the reliability of results using the noninvasive measurement of aortic blood flow (ABF) (combined Doppler- and M-Mode transesophageal ultrasound, Dynemo 3000, Sometec Inc, Paris, France). METHODS: In 75 patients during cardiac or major abdominal surgery we performed 313 simultaneous measurements of CO and ABF. RESULTS: Placement of ultrasound probe into correct position took less than 2 min. Quality and stability of ultrasound signals were good. The coefficient of correlation between ABF and CO was found to be 0.89 with CO = 0.97 x ABF + 1.1, Bland-Altman-Test positive. CONCLUSION: Results of ABF detected by combined Doppler- and M-Mode Echography are comparable with results of CO obtained by PAC. Therefore we are convinced that this noninvasive method will find its place in clinical situations of compromised CO. PMID- 10788991 TI - [DIVI emergency medicine protocol, version 4.0]. PMID- 10788992 TI - [Aloys Martin (1818-1891) "Trailblazer of anesthesia". An addendum to his life and influence]. AB - OBJECTIVES: Alloys Martin has recently been called "a pioneer of surgical anaesthesia in Germany" [41]. This gave rise to an investigation into further aspects of his professional and private biography and the social as well as political context. METHODS: Documents in archives and relevant medical and historical literature relating to the objectives were surveyed and analyzed. RESULTS: In 1848 the anaesthesiological articles Martin had been contributing came to an end. He then worked as a university lecturer and professor at the University of Munich, as an official forensic and charity physician as well as in medical practice in Munich. Among other things, he published diverse articles on the schematism of Bavarian civilian and military physicians, on infectious diseases and edited scientific journals. As a founder and editor of the Bayerisches arztliches intelligenzblatt (Bavarian Medical Information Newspaper), renamed the Munchner Medizinische Wochenschrift (Munich Medical Weekly Journal) in 1886, he guided the publication to a central position in a medical community just developing professional status. Martin was also the founder and for many years the chairman of societies for voluntary medical assistance to the poor and the Froebel kindergartens in Munich. Although he himself was of lower class origin and under pressure from conservative circles at the university and ministry due to his liberal political views, his life and works bear witness to his own effective emancipation and rise into the "Bildungsburgertum" (educated class) of the 19th century. CONCLUSIONS: Alloys Martin is a historical figure in anaesthesiology. In his function of promoting the professionalization of the whole Bavarian medical community, he also ranks highly. In view of his self abnegating medical service devoted to the common good, he may still today be considered an example. PMID- 10788993 TI - [Prevention of nosocomial infections: intensive care units under the magnifying glass. The first german project for distinguishing between preventable and unpreventable nosocomial diseases]. PMID- 10788994 TI - [Is a score for prediction of emesis after pelvic laparoscopy possible? Remarks on a paper in Der Anaesthesist (1999) 48:705-712]. PMID- 10788995 TI - [Stomach-intestine atonia]. PMID- 10788996 TI - [Adverse effects of angiotensin receptor antagonists]. PMID- 10788997 TI - [Platelet aggregation in spinal anesthesia]. PMID- 10788998 TI - [Complications in the recovery room]. PMID- 10788999 TI - Changes in EEG power spectra during biofeedback of slow cortical potentials in epilepsy. AB - The goal of the study was to explore parallel changes in EEG spectral frequencies during biofeedback of slow cortical potentials (SCPs) in epilepsy patients. Thirty-four patients with intractable focal epilepsy participated in 35 sessions of SCP self-regulation training. The spectral analysis was carried out for the EEG recorded at the same electrode site (Cz) that was used for SCP feedback. The most prominent effect was the increase in the theta 2 power (6.0-7.9 Hz) and the relative power decrement in all other frequency bands (particularly delta 1, alpha 2 and beta 2) in transfer trials (i.e., where patients controlled their SCPs without continuous feedback) compared with feedback trials. In the second half of the training course (i.e., sessions 21-35) larger power values in the delta, theta, and alpha bands were found when patients were required to produce positive versus negative SCP shifts. Both across-subject and across-session (within-subject) correlations between spectral EEG parameters, on the one hand, and SCP data, on the other hand, were low and inconsistent, contrary to high and stable correlations between different spectral variables. This fact, as well as the lack of considerable task-dependent effects during the first part of training, indicates that learned SCP shifts did not directly lead to the specific dynamics of the EEG power spectra. Rather, these dynamics were related to nonspecific changes in patients' brain state. PMID- 10789000 TI - EMG reactivity and oral habits among facial pain patients in a scheduled-waiting competitive task. AB - For individuals with temporomandibular disorder (TMD) it has been theorized that stressful events trigger oral habits (e.g., teeth grinding), thereby increasing masticatory muscle tension and subsequent pain. Recent research involving adjunctive behaviors found an increase in masseter surface EMG (sEMG) and oral habits when students with TMD symptomatology were placed on a fixed-time reinforcement schedule. The current study used a treatment-seeking community sample with TMD symptomatology in a competitive task designed to be a more naturalistic Fixed Time task. The experiment consisted of Adaptation, Free-Play, Scheduled-Play, and Recovery phases. During the Scheduled-Play phase participants played, and waited to play, an electronic poker game. Results indicated that masseter muscle tension in the Scheduled-Play phase was significantly higher (p < .001) than in any other phase. Moreover, during the Scheduled-Play phase masseter sEMG was higher (p < .001) when participants waited to play. Self-reported oral habits and overall affect were significantly higher (p's < .05) in the Free-Play and Scheduled-Play phases relative to Adaptation and Recovery. The observation that masseter sEMG was elevated during the Scheduled-Play phase relative to all other phases, and within the Scheduled-Play phase sEMG was highest while waiting, suggests that adjunctive oral habits may lead to TMD symptomatology. PMID- 10789001 TI - Religious coping with chronic pain. AB - This study examined the role of religious and nonreligious cognitive-behavioral coping in a sample of 61 chronic pain patients from a midwestern pain clinic. Participants described their chronic pain and indicated their use of religious and nonreligious cognitive-behavioral coping strategies. Results supported a multidimensional conceptualization of religious coping that includes both positive and negative strategies. Positive religious coping strategies were associated significantly with positive affect and religious outcome after statistically controlling for demographic variables. In contrast, measures of negative religious coping strategies were not associated significantly with outcome variables. Several significant associations also were found between nonreligious cognitive-behavioral coping strategies and outcome variables. The results underscore the need for further research concerning the contributions of religious coping in adjustment to chronic pain. Practitioners of applied psychophysiology should assess their chronic pain patients' religious appraisals and religious coping as another important stress management strategy. PMID- 10789002 TI - Reliability of infrared plethysmography in BVP biofeedback therapy and the relevance for clinical application. AB - Biofeedback methods are well established as behavioral techniques for the therapy of various psychophysiological diseases. The forms of feedback generally employed are muscle activity (electromyogram), skin temperature, brain activity (electroencephalogram), and vasomotoricity. The latter technique, which employs plethysmographic feedback, has been studied most extensively in the therapy of migraine (vasoconstriction training, blood volume pulse training). Although the clinical efficacy has been demonstrated in several studies, little is known about the psychometric properties of this technique. This study examined the intrasession and intersession reliability of the pulse volume amplitude (PVA). The results showed that the PVA measurements within a single biofeedback session were highly reliable. Repositioning of the probe within the session resulted in a lower correlation coefficient, but one that was still sizable and significant. The PVA values from different sessions were not reliable (or comparable). PMID- 10789003 TI - Specific muscle EMG biofeedback for hand dystonia. AB - Currently available therapies have only limited success in patients having hand dystonia (writer's cramp). We employed specific muscle EMG biofeedback (audio feedback of the EMG from proximal large muscles of the limb that show abnormally high activity during writing) in 10 of 13 consecutive patients (age, 19-62 years; all males) with a duration of illness from 6 months to 8 years. In three patients, biofeedback was not applicable due to lack of abnormal EMG values. Nine patients showed dystonic posture during writing and had hypertrophy of one or more large muscles of the dominant hand. The remaining four patients showed either involvement of small muscles or muscle wasting. Ten patients were given four or more sessions of EMG audio biofeedback from the proximal large limb muscles, which showed maximum EMG activity. They also practiced writing daily with the relaxed limb for 5 to 10 min. Nine patients showed improvement from 37 to 93% in handwriting, alleviation of discomfort, and pain (assessed on a visual analogue scale). One patient did not show any improvement. Thus EMG biofeedback improved the clinical and electromyographic picture in those patients with hand dystonia who showed EMG overactivity of proximal limb muscles during writing. This specific type of EMG biofeedback appears to be a promising tool for hand dystonia and might also be applied to other types of dystonias. PMID- 10789004 TI - [Is autopsy still usefull?]. PMID- 10789005 TI - [Immunomorphological study of diabetic microangiopathy]. AB - Femoral skin biopsies of 300 patients with insulin-dependent and non-insulin dependent diabetes were studied immunomorphologically and the results were compared to the level of immunoglobulins in the blood serum. It is found that IgG, beta-lipoproteins, complement were registered in the arterioles and capillary walls in diabetic microangiopathy more frequently than in healthy persons. The conclusion is made that in development of diabetic microangiopathy, in both types of diabetes, immune (immunocomplex) mechanisms that damage vascular wall are involved. PMID- 10789006 TI - [Dynamic of morphological changes of cerebral hematomas induced by arterial hypertension (analysis of surgical and autopsy materials)]. AB - The pattern of morphological changes in blood clots, brain tissue and vessels of hypertensive hematomas (CH) was studied in 79 surgical and autopsy cases. Brain tissue compression degeneration of cells and fibers, edema were observed during the first 6 hours after the onset of hemorrhage. Necrosis and marked leukocyte infiltration appeared to progress gradually and reach maximum by the disease day 2-3. The degeneration manifested by formation of gliomesodermal capsule surrounding the haemorrhagic lesion from day 3 to 8. Dilatation of small cerebral vessels was followed by an increase in permeability and necrosis of vascular walls predominated within 24-72 hours after initial bleeding. It is suggested that the first 24 hours after CH onset are optimal for surgery. PMID- 10789007 TI - [Comparative ultrastructural analysis of capillaries in the conducting and contracting myocardium of the heart sino-auricular region in suddenly died patients with coronary artery disease and alcoholic cardiomyopathy]. AB - The EM study of capillaries in the sinus node (SN) and adjacent working myocardium has been performed in 12 patients who ceased a sudden coronal cardiac death at the age 34-69 years and in 8 patients with alcoholic cardiomyopathy who suddenly died at the age 32-60 years. Quantitative evaluation of the density of the capillaries and their diameters in the myocardium of this region was made. Acute and chronic changes were observed in the capillaries of SN and working myocardium. Possible causes of the ultrastructural pathology of the microvascular system in the sinoauricular region of the heart in persons who died suddenly are discussed. PMID- 10789008 TI - [Comparative diagnostic value of the analysis of the skeletal muscle and lymphocytes in mitochondrial diseases]. AB - Cytochemical analysis of lymphocytes' enzymes may could be an alternative method to skeletal muscle biopsy in diagnosis of mitochondrial pathology. We investigated biopsies of skeletal muscles and cytochemical status of lymphocytes in 14 children with mitochondrial pathology. Lymphocytes were investigated also in 12 health children. The data on mitochondrial state in the skeletal muscles and peripheral blood lymphocytes were comparable. The less were the size and functional activity of single mitochondria in lymphocytes, the higher was mitochondrial insufficiency in the skeletal muscle. Enzymatic status of peripheral blood lymphocytes is not less indicative of mitochondrial insufficiency in children than mitochondrial characteristics in skeletal muscles. It reflects multisystem mitochondrial insufficiency. The results obtained are determined by both mitochondrial pathological modifications and their compensatory response to pathological process. PMID- 10789009 TI - [Intraductal breast cancer]. AB - 186 cases of intraductal carcinoma have been analyzed. Twelve morphological variants and three degrees of differentiation are distinguished. Different results of the treatment of patients with the same clinical diagnosis "carcinoma in situ" may be explained by heterogeneity of intraductal carcinoma. PMID- 10789010 TI - [Immunohistochemical study of breast neoplasms cells using different markers of proliferation]. AB - Proliferative activity was studied in 58 breast tumors: 10 benign tumors and 48 carcinomas 41 of which were invasive ductal carcinomas. Combined use of 3 proliferation markers (5-bromo-2-deoxyuridine, nuclear antigen of proliferating cells and Ki-67 antigen) which reflect different phases of mitotic cycle, enable determination of the S-phase index and estimation of the tumor proliferative pool as well as percentage of cells in S-phase and in mitosis and, respectively, relative duration of these phases. In breast carcinoma, S-phase is the longest phase of the mitotic cycle (45% of its duration), mitosis takes 5%. Proliferation indices when all three markers were used carry high positive correlation (r = 0.73-0.94; p < 0.000001). Proliferative activity of breast carcinoma is 5 times higher than in benign tumors and significantly increases in recurrent tumors and with growing histological malignancy of the tumor. A positive correlation exists between proliferative activity of the primary carcinoma and that of metastatic tumor in regional lymph nodes but proliferation in metastasis is, as a rule, less active than in the primary tumors. Antigen Ki-67 is the most optimal among the above proliferation markers because its expression reflects the tumor proliferative pool and its use does not require additional procedures and calculation of proliferating cells is simple and reliable. PMID- 10789011 TI - [Immunohistochemical analysis of cyclin D1 in cervical adenocarcinoma]. AB - Cyclin D1 expression in various types of uterine cervix adenocarcinoma is studied immunohistochemically. Cyclin D1 location in the nucleus and cytoplasm is revealed and its dependence on the degree of tumor differentiation. PMID- 10789012 TI - [Pathogenetic characteristics of late spontaneous miscarriage in ascending infection of amnion: reaction of placenta]. AB - Results of pathomorphological examination of placenta in 262 cases of late spontaneous miscarriages produced by ascending infection of fetal liquids with opportunistic microflora are presented. Sequential involvement in the exudative inflammatory process of the fetal membranes was observed (placenta and umbilical cord) depending on the term of pregnancy and duration of placenta contact with the infected fetal liquids. Exudative funiculitis was observed not infrequently this indicating direct involvement of the fetus in the intrauterine infectious process. PMID- 10789013 TI - [Experimental-morphological and clinical results of laser destruction of adenohypophysis]. AB - Feasibility of focal adenohypophysis destruction by Nd:YAG laser is experimentally proven. The destruction is not followed by considerable damage of other structures of the hypothalamus-hypophysis region. It is established that the focus of coagulation necrosis produced by laser radiation is replaced by scar tissue. A surgical treatment of hypophysis adenoma surgical treatment is proposed. Its effectiveness, safety and low traumatism were confirmed at its clinical trial. PMID- 10789014 TI - [Basic principles and new nosological forms of the second edition of the histological classification of soft tissue tumors (update of 1994)]. AB - General principles of the second review of the soft tumours classification and brief description of nosological unities absent in the first edition are presented. PMID- 10789015 TI - [Etiological and nosological structure of liver diseases (on autopsy data of clinics of I.M. Sechenov Moscow Medical Academy in 1988-1997)]. AB - 4908 autopsy protocols were analysed and liver pathology was found in 1.7% of cases. Alcoholic and viral diseases dominated among liver pathology cases, steatosis among alcoholic damages, cirrhosis among viral diseases, primary biliary cirrhosis among autoimmune diseases and hepatocellular carcinoma among malignant tumors. Comparison of the two decades (1988-1997 and 1978-1987) revealed certain growth of alcoholic and viral diseases and their possible combinations. The number of liver tumors of viral etiology also went up. PMID- 10789016 TI - [Modification of pathologo-anatomic method of heart dissection after valve prosthesises implantation]. AB - Modification of the method of heart dissection in patients who died after valve replacement is suggested. After standard atrial cutting dissection of the ventricles is made from the apex along the interatrial fissure without cutting the proximal regions of the coronary arteries. The method allows good inspection of the ventricular cavities and elucidation of the relationships between anatomical structures and artificial valves. PMID- 10789017 TI - [P.P.Zobolotnov--founder of the Saratov research school of pathologists]. PMID- 10789018 TI - [60 years anniversary of the pathology service in the Chelyabinsk region]. PMID- 10789019 TI - [Inflammation mediators in pathogenesis of infection-toxic shock of meningococcal etiology]. AB - Up-to-date advances made in understanding of the events underlying meningococcal infection-toxic shock (ITS) are reviewed. Endotoxin properties, its interaction with transport protein and specific membrane receptors entailing activation of macrophages and secretion of cytokines TNF and IL-1 are described. Both of them are considered as major mediators of ITS. Recent information about activation of complement and coagulation cascades, changes of kinetic and functional properties of polymorphonuclear leukocytes are summarized. As to secondary mediators, the emphasis is placed on bioregulatory system L-arginine-NO. A scheme of pathogenesis of meningococcal ITS is provided. The latter is corresponding both clinically and morphologically to other endotoxin shocks and its peculiarities in excessive activation of coagulation system. PMID- 10789021 TI - Intrusive thoughts and psychological distress among breast cancer survivors: global meaning as a possible protective factor. AB - Previous research has consistently demonstrated a positive association between intrusive thoughts about stressful experiences and psychological distress. The strength of this relation, however, has varied considerably across studies. To examine the possibility that an individual's sense of global meaning (i.e., the existential belief that one's life has purpose and order) may moderate the relation between intrusive thoughts and psychological distress, the authors conducted telephone assessments of 61 women who had survived breast cancer. Results confirmed that the frequency of intrusive thoughts was positively related to psychological distress. Global meaning, moreover, moderated the relation between intrusive thoughts and psychological distress consistent with the authors' hypotheses. Among women with lower global meaning, more frequent intrusive thoughts were associated with higher psychological distress. No association was found between intrusive thoughts and psychological distress among those participants with higher global meaning. PMID- 10789020 TI - High versus low basal cortisol secretion in asymptomatic, medication-free HIV infected men: differential effects of severe life stress on parameters of immune status. AB - The authors hypothesized that HIV-infected men with high basal cortisol secretion would exhibit greater stress-related reductions in the ratio of Th1/Th2 cell derived cytokines and numbers of CD8+ T and NK lymphocytes than low basal cortisol secretors. A semistructured interview was used to assess life stress during the preceding 6 months of 94 HIV-infected men classified as high and low cortisol secretors (n = 47/group). Increased levels of severe life stress were highly correlated with lower numbers of CD8+ T cells, CD16+ and CD56+ NK cells, CD57+ cells, and higher DHEA-S concentrations in the high cortisol group. Conversely, no significant correlations were found in the low cortisol group. No correlations were found between stress and CD4+ T helper/inducer cell counts, cytokine production, or testosterone levels in either participating group. These data suggest that severe stress in combination with high glucocorticoid activity may modify select parameters of immune status in HIV-infected men. PMID- 10789022 TI - Self-report sleep habits as predictors of subjective sleepiness. AB - Although sleepiness is pervasive in our society, there is little agreement on how to measure sleepiness or on how well sleepiness is actually related to sleep habits. To better assess how subjective sleepiness is related to sleep, the authors used self-report measures of sleep quantity, sleep quality, and napping to predict 4 different sleepiness-related measures in a group of healthy young and middle-aged-to-older adults. A forward regression analysis indicated that sleep quality was better than sleep quantity as a predictor of participants' sleepiness. The sleep measures, furthermore, predicted sleepiness better in the older adults than in the younger adults. Finally, the 4 sleepiness measures differed in how well they were related to sleep. The findings in the study suggest that sleepiness is a complex phenomenon rather than a simple reflection of sleep quantity. PMID- 10789023 TI - The development and validation of the Sense of Support Scale. AB - Two studies were conducted to develop and validate the Sense of Support Scale (SSS). Study 1, which consisted of scale development and an evaluation with a sample of corporate and university employees, supported the scale's internal consistency and construct validity. Study 2, which was conducted with a sample of undergraduate students, was designed to evaluate the revised and shortened version of the SSS. The scale was found to be internally consistent and had a high test-retest reliability. Concurrent validity was supported by significant correlations to the Social Provisions Scale and the Interpersonal Support Evaluation List. Construct validity was supported by significant positive correlations to hardiness and approach-coping and significant negative correlations to avoidance-coping, stress, and symptoms of illness. The relationship between the SSS and symptoms of illness supported a main effect rather than a stress-buffering model. Implications for using the SSS to investigate the influences of social support on health are discussed. PMID- 10789024 TI - Against methodolatry: some comments on Elliott, Fischer, and Rennie. PMID- 10789025 TI - The timing, triggers and qualities of recovered memories in therapy. AB - OBJECTIVES: To report descriptive data on memory recovery of traumatic material including: degree of prior amnesia, triggers to recovery, qualities of the memory and length of time taken to recover different types of memory. DESIGN: British Psychological Society practitioners who reported having clients with recovered memories in a previous large-scale survey were contacted for an indepth interview. They each provided information on up to three such clients. METHODS: From telephone interviews with 108 therapists, we elicited 236 detailed accounts of clients recovering a traumatic memory. Interviews were semistructured with investigator-based ratings. RESULTS: The degree of amnesia varied widely according to therapists: some clients had prior total amnesia, others a prior vague sense or suspicion, and others prior partial memories. Time in therapy before first recall was longer for memories involving child sexual abuse than for memories of other traumas. The majority of the memories, but not all, were similar to those reported by patients with post-traumatic stress disorder: they were fragmented, accompanied by high levels of emotion, and experienced as a reliving of the original event. CONCLUSIONS: On the basis of current understanding of memory processes, several mechanisms may be needed to explain all the data, including disruptions to the encoding and retrieval of traumatic events and errors in the attribution of the source of specific memories. It will be important to confirm these findings by interviewing clients themselves. PMID- 10789026 TI - Subjective improvement in PTSD patients with treatment by imaginal exposure or cognitive therapy: session by session changes. AB - A self-rating scale, the Subjective Symptom Checklist (SSC) was developed to assess PTSD patients' perception of their symptoms between treatment sessions. Reliability and validity of the scale were acceptable. The scale was used in a treatment trial comparing the efficacy of cognitive therapy versus imaginal exposure. Conventional pre-post treatment comparisons using standardized assessments showed no differences between the two treatments. However, when patients who failed to respond to treatment were excluded, group treatment effects became apparent. Patients who received imaginal exposure showed a significantly greater reduction in subjective ratings of their symptoms than did those who received cognitive therapy. It was cautiously concluded that although some PTSD patients could not tolerate exposure, those who could may receive greater subjective benefit than those who received cognitive therapy. PMID- 10789027 TI - Prevalence and predictors of post-traumatic stress symptoms following childbirth. AB - OBJECTIVES: To identify the prevalence and potential predictors of post-traumatic stress type symptoms following labour. DESIGN: A large sample, within participants design with initial assessment and postal follow-up was utilized. METHOD: Two hundred and sixty-four women who had 'normal' births were assessed within 72 hours on potential predictive measures and at 6 weeks post-partum for levels of symptoms of intrusions, avoidance and hyperarousal on a questionnaire derived from DSM-IV (American Psychiatric Association, 1994) criteria. Symptoms of depression and anxiety were also assessed. RESULTS: Three per cent showed questionnaire responses suggesting clinically significant levels on all three post-traumatic stress dimensions and a further 24% on at least one of these dimensions. Forward stepwise regression analysis yielded models for predicting outcome variables. Perceptions of low levels of support from partner and staff, patterns of blame and low perceived control in labour were found to be particularly related to experience of post-traumatic stress symptoms. Personal vulnerability factors such as previous mental health difficulties and trait anxiety were also related to such symptoms as well as being relevant predictors for anxiety and depression. CONCLUSIONS: A proportion of women reports all three aspects of post-traumatic stress type symptoms following childbirth with many more reporting some components. A broader conceptualization of post-partum distress which takes account of the impact of labour is required. There may be opportunities for prevention through providing care in labour that enhances perceptions of control and support. PMID- 10789028 TI - Psychological management of post-stroke depression. AB - PURPOSE: To evaluate existing research literature on psychological interventions to manage post-stroke depression (PSD). METHOD: First, the particular nature of post-stroke depression (PSD) was established. Then, relevant published material was identified through computerized literature searches (to May 1999) and via the first author's clinical experience in the area. This material was critically reviewed against recommended standards for the empirical validation of treatment effectiveness. RESULTS: Methodological limitations in existing research prevent a conclusion as any one psychological intervention has empirical support for its effectiveness. However, cognitive behaviour therapy in particular was identified as worthy of further investigation. CONCLUSIONS: It is recommended that future research investigates treatments for PSD appropriate for those with cognitive impairment and communication difficulties, younger versus older stroke victims and those in institutional settings. Studies should also consider the necessity of specialist assessment strategies and allow for possible subtypes of PSD for which psychological treatments might be differentially effective. PMID- 10789029 TI - Cognitive factors in predisposition to auditory and visual hallucinations. AB - OBJECTIVES: This study adapted the Launay Slade Hallucination Scale (LSHS) to measure predisposition to auditory and visual hallucinations and examine the relationship between meta-cognition and predisposition in a non-psychiatric population. It also tested the hypothesis that individuals highly predisposed to hallucination would show positive and negative meta-cognitive beliefs and report the use of different thought control strategies. DESIGN: A within participants correlational design was employed. METHODS: A revised LSHS was administered to 105 normal participants who were also asked to complete questionnaires assessing paranoia, meta-cognitive beliefs, thought control strategies, anxiety, depression and beliefs about unusual perceptual experiences. RESULTS: Two empirically distinct but correlated hallucinatory traits (auditory and visual) were measured by the modified LSHS. Consistent with predictions, it was found that positive beliefs about unusual perceptual experiences were the best predictor of predisposition to auditory and visual hallucinations and that those participants who scored higher on predisposition to hallucination used different thought control strategies and had different negative meta-cognitive beliefs in comparison with participants of low predisposition. CONCLUSIONS: Meta-cognitive beliefs about thoughts and hallucinatory phenomena appear to be implicated in predisposition to hallucination. The theoretical and clinical implications of the findings are discussed. PMID- 10789030 TI - A confirmatory factor analysis of the Hospital Anxiety and Depression scale: comparing empirically and theoretically derived structures. AB - OBJECTIVES: To compare the fit of various factor solutions for the Hospital Anxiety and Depression scale (HAD; Zigmond & Snaith, 1983). DESIGN: A cross sectional postal survey was used to collect the data from community-based participants in the West of Scotland Twenty-07 study. METHODS: The HAD scale, a 14-item self-administered measure of anxiety and depression, was completed by 2547 participants from three age cohorts (aged approximately 18, 39 and 58 years). Using confirmatory factor analyses four models suggested by prior exploratory factor analyses were compared to a model derived from Clark and Watson's (1991) tripartite theory of anxiety and depression. RESULTS: The model derived from the tripartite theory of anxiety and depression (with factors labelled negative affectivity, anhedonic depression and autonomic anxiety) produced the closest fit to the data. This model produced a good fit in all three cohorts although group comparisons suggested that there were variations in the strength of some factor loadings across the three age groups. A model that had a hierarchical arrangement of the three factors in the tripartite model was also produced. This model fit the data equally as well as did the 'flat' tripartite model. CONCLUSIONS: Three factors appear to underlie the HAD scale. Research is needed that examines whether or not using sub-scales based on these factors increases the ability of the HAD scale to detect cases of anxiety and depression. PMID- 10789031 TI - The validity of the NART-RSPM index in detecting intellectual decline following traumatic brain injury: a controlled study. AB - The present study replicated Van den Broek and Bradshaw's (1994) study by developing a multiple regression equation to predict Ravens Standard Progressive Matrices (RSPM) scores from participants' National Adult Reading Test (NART) scores and demographic variables. A test of the validity of this procedure was undertaken by applying both the Van den Broek and Bradshaw equation and the present study's equation to a mixed sample of orthopedic control (OC) and closed head injury (CHI) participants. Premorbid RSPM scores were estimated using both equations and then compared to obtained scores. Results indicated that both equations had good predictive accuracy in estimating RSPM scores and were able to detect significantly higher rates of impairment in general intelligence in the CHI participants (24%) compared to the OC sample (7%). PMID- 10789032 TI - Bone graft substitutes in spine surgery. PMID- 10789033 TI - Posterior osteotomies with pedicle substraction for flat back and associated syndromes. Technique and results of a prospective study. PMID- 10789034 TI - Computerized frameless stereotactic image-guided spinal surgery. PMID- 10789035 TI - Endoscopic thoracoplasty and anterior spinal release in scoliotic deformity. AB - Severe rigid and complex deformities of the spine often require a first stage procedure to adequately mobilize the spine to provide adequate flexibility during the actual application of instrumentation for surgical correction. This first stage involves soft tissue releases and removal of intervertebral disks from the anterior spine. Exposure to the anterior spine has traditionally been accomplished through thoracotomy or a thoracolumbar incision. The open thoracotomy, however, has several disadvantages, such as post-thoracotomy pain, a large scar, and breathing difficulties. Since the first thoracoscopy at the beginning of this century, the procedure, at first limited by the available technology, has gradually gained more uses and favor among physicians. The last 10 years have seen significant improvement in optical technology and instrumentation. These advances have allowed the spine surgeon to begin performing anterior spinal releases using endoscopic techniques. These endoscopic techniques can carry the same efficacy as the open thoracotomy but less morbidity. Thoracoplasty, the resection of rib segments, is an excellent way to not only correct the convex rib deformities seen in scoliosis but it is also an excellent source of bone which can be used as a graft for fusion. Although traditionally, thoracoplasty has also been done through an open procedure, it can be performed endoscopically. This prospective study presents nine patients who underwent combined endoscopic anterior spinal release and thoracoplasty followed by same day posterior instrumentation and fusion for correction of their spinal deformities. All nine procedures were completed successfully endoscopically. It is our conclusion that in the hands of an experienced surgeon, the endoscopic technique is an excellent procedure providing the same efficacy as the open thoracotomy. There is however a learning curve associated with the procedure. In addition, a team approach in which the surgeon and an experienced anesthesiologist with experience with double lumen intubation and selective single lung ventilation and thoracoscopic surgery is crucial. Although there were no surgical complications related to the thoracoscopic technique one patient did require prolonged intubation postoperatively, which leads us to believe that single lung ventilation in and of itself is very demanding and each patient must be considered carefully prior to its undertaking. PMID- 10789036 TI - Clinical efficacy of lumbar and lumbosacral fusion using the Boucher facet screw fixation technique. AB - The clinical outcome of posterolateral lumbar and lumbosacral fusions with facet screw fixation using the Boucher technique is reviewed retrospectively. The Boucher technique uses AO cortical screws that traverse the facet joint at an angle from the superior lamina, in a medial to lateral direction, to enter the respective inferior pedicle. The procedure was performed on 57 consecutive patients who had a degenerative disk disease. The pre-operative diagnosis was based on a physical examination and radiographs with myelography performed when deemed necessary. Surgery had been indicated and performed on patients with unresolved pain after 6 months of conservative therapy; radiographic signs of disk space narrowing, facet arthrosis, degenerative spondylolisthesis or instability on flexion and extension views. No prior history of spinal surgery, no involvement in any litigation or Workers Compensation process, and no detected psychiatric disturbances were also inclusion criteria for this study. Eighteen consecutive male and 39 consecutive female patients were included in the study. The average age at the time of surgery was 41.8 years. The mean active clinical follow-up (office visits) time was 5.06 months with the minimum follow-up time of 3 months. Two years after their surgery, all the patients were asked to fill out a questionnaire from which their progress was evaluated. All questionnaires were completed and returned. Postoperatively, patients were placed in either a lumbosacral corset, short rigid lumbosacral molded polypropylene brace or body cast. Overall clinical results for single level fusions showed that 91.2% of patients (31 of 34) had excellent results and 8.8% (3 of 34) had poor results. Of the multiple level fusions, 86.4% of patients (19 of 22) had excellent or good results and 9.1% (2 of 22) had poor results. Post-operative complications included only three wound infections at the iliac crest donor site. This study demonstrates that facet screw fixation using the Boucher technique has advantages. The technique not only is easy to implement by placing a small screw through a facet joint and it's respective pedicle, but produces excellent clinical results that are comparable to the other more bulky spinal instrumentation systems. PMID- 10789037 TI - Spinal infections. Diagnosis and treatment. AB - Spinal infections encompass a broad spectrum of pathology. Any element of the spine, the vertebrae, disks, surrounding soft tissues, potential spaces, or the cord itself may be involved solely, or more commonly, in combination. Staphylococcus aureus is the organism most frequently found, with Gram negative bacteria often cited in association with infections of the urinary tract and in intravenous drug abusers. Early diagnosis and treatment are essential for optimal outcomes. In some cases, the only necessary treatment consists of parenteral antibiotics and immobilization. However, certain instances require operative intervention. The goals of treatment for any infection of the spine are: early diagnosis with identification of the offending organism, preservation of neurological function or cessation of progressive neurologic deficit, and complete eradication of the infection with a pain-free, stable spine. PMID- 10789038 TI - Internal fixation of cervical trauma following corpectomy and reconstruction. The effects of posterior element injury. AB - Although biomechanical data indicates that anterior fixation alone in unstable cervical injuries may not provide adequate stability, reports of clinical series indicate general success with this method of treatment. The specific contribution of posterior column injury to overall stability following reconstruction has not been evaluated. This study examined the biomechanical stability of anterior and/or posterior plate fixation following anterior corpectomy and reconstruction for unstable cervical injuries with varying degrees of posterior element injury. The C4-C6 motion segments of ten fresh frozen bovine cervical spines were used. After mounting, nondestructive mechanical testing in axial compression, torsion, flexion, extension, and lateral bending was done as an intact control. A C5 corpectomy with reconstruction using a synthetic bone graft was performed and the posterior ligaments sectioned at the C5-C6 level. Each specimen was sequentially instrumented with anterior and posterior plating alone and in combination and each construct was mechanically retested. The specimens were then further destabilized by bilateral facetectomies at C5-C6 and again tested with the same instrumentation combinations. In comparison to the controls, the spines with a C5 corpectomy/bone graft and posterior ligament rupture with anterior plating demonstrated significantly increased stiffness in flexion, extension, and lateral bending; posterior plating increased stiffness in only flexion and lateral bending. In axial compression and torsion, anterior or posterior plating demonstrated stiffness similar to the controls. Further destabilization by facetectomy significantly decreased stiffness of the instrumented construct (less than control) in torsion with anterior or posterior plate fixation alone. Combined plating showed increased stability compared to controls in all loading conditions for both patterns of instability. Anterior plating alone was able to restore the stability of the cervical spines with posterior ligamentous injury after corpectomy, but it failed to do so with the addition of bilateral facetectomies. For the unstable cervical spine with significant bilateral loss of posterior bony contact, anterior or posterior plating alone may not provide sufficient stabilization in the absence of any additional external immobilization. Combined plating should be considered, which may obviate the need for external immobilization. PMID- 10789039 TI - Traumatic rotatory displacement of the lower cervical spine. AB - The authors describe the classification for traumatic rotary injuries of the cervical spine. The classification is based on a review of 306 severe lower cervical spine injuries observed in 255 patients between 1980 and 1994. Traumatic rotatory displacements (TRD) represented 39% of the 306 severe injuries. Three different lesions were observed: unilateral facet fractures, fracture-separation of the articular pillars, and unilateral facet dislocations. PMID- 10789040 TI - [Changes in incidence and prevalence of acute and chronic pancreatitis in Germany]. AB - It is difficult to provide valid data with regard to changes in incidence and prevalence of acute pancreatitis in Germany. The lack of information is due to strict legal regulations to protect personal data, a certain lack of interest in epidemiological research and the lack of exact statistics of diseases treated in hospitals. The incidence of acute pancreatitis in other industrialized Western countries is about 10 new cases per year and 100,000 inhabitants. The increase of alcohol consumption over the last 30 years is associated with an increase of ethanol-induced pancreatitis. One may speculate that due to improved diagnostic possibilities, such as CT scan, cases where acute pancreatitis is diagnosed at autopsy are rare. Due to the increase in alcohol consumption it can be assume that the incidence and prevalence of chronic pancreatitis have increased in Germany similar to well-documented data from Denmark. Since alcohol consumption is slightly decreasing in Germany it is likely that the present incidence of about 10 cases of chronic pancreatitis per 100,000 inhabitants will also decrease. PMID- 10789041 TI - [New pathophysiologic knowledge about acute pancreatitis]. AB - Induction of acute pancreatitis follows a uniform mechanism independent of the different etiologic factors such as gallstones, alcohol, ischemia, hyperlipidemia, hypercalcemia, hereditary and others. Each cause seems to affect primarily the acinar cell, resulting in premature intracellular activation of trypsinogen and other digestive enzymes. Activated enzymes and oxygen free radicals injure the acinar cell and cause a release of cytokines and vasoactive mediators, attract inflammatory cells and activate the vascular endothelium as well as the expression of adhesion molecules. The disturbance of the pancreatic microcirculation induces a progression from edematous to necrotizing pancreatitis independent of the early intracellular events, including protease activation. Specific therapy must be directed towards microperfusion failure as a secondary pathogenetic step, since the initial enzyme activation and cytokine release is irreversible by the time of clinical presentation. In experimental designs comparable to the clinical situation the following therapeutic principles have proven beneficial: increase of blood fluidity by dextran, inhibition of leukocyte endothelium interaction by ICAM-1 antibodies, and blockade of local vasoconstriction by endothelin-receptor antagonists. PMID- 10789042 TI - [Acute pancreatitis. Early elective endoscopic intervention]. AB - Early endoscopic intervention is required in the treatment of biliary pancreatitis in the presence of cholangitis or jaundice, possibly with dilated common bile duct. Also patients with predicted severe disease, lack of spontaneous clinical improvement or progressive jaundice might benefit from endoscopic retrograde cholangiopancreatography (ERCP) with sphincterotomy. If endoscopy is indicated, it should be performed as soon as possible. Injection of contrast medium into a dilated bile duct is accompanied by the risk of cholangitis. Therefore therapeutic decompression by sphincterotomy and stone extraction is necessary, or, if this is not possible, insertion of a naso-biliary tube for drainage. Prophylactic use of antibiotics is recommended periinterventionally. Early endoscopic intervention is not necessary in patients with mild biliary pancreatitis and spontaneous clinical improvement. Here, ERCP can be performed later, if indicated. PMID- 10789043 TI - [Parenchymal necrosis: infection and other indications for debridement and drainage]. AB - In the present review we describe our experience with debridement in necrotizing pancreatitis, with particular emphasis on the indications for surgery and the outcome in patients with infected and sterile necrosis. Within 7 years 64 patients with necrotizing pancreatitis underwent surgery with a median preoperative APACHE II score of 9. The indications for surgery were either proven infection or persistence of symptoms. Thirty-six patients were found to have infected necrosis, while 28 patients had sterile necrosis. There was no significant difference between the two groups regarding pre- and postoperative parameters. The surgical technique involved a blunt necrosectomy followed by closed packing with Penrose drains. This technique produced the lowest reported mortality of 6.2% with a reoperation rate of only 17%, demonstrating that even with sterile necrosis surgical management can be carried out safely. PMID- 10789044 TI - [Necrosectomy or anatomically guided resection in acute pancreatitis]. AB - Necrotizing pancreatitis is still associated with considerable morbidity and mortality. Formerly, surgical treatment with early and extensive pancreatic resection has been the standard. Improvements in our understanding of the pathogenesis of the disease and progress in the field of intensive care therapy have made conservative therapy the initial standard in the treatment of necrotizing pancreatitis. A considerable percentage of patients with sterile necrosis can be managed with low morbidity and mortality by conservative treatment without operation. Nevertheless, surgical treatment is indicated in patients with infected necrosis and in sterile necrosis if multiorgan failure persists over a limited period of time despite maximum intensive care therapy. Pancreatic resection should be abandoned for surgical treatment of pancreatic necrosis, as results with regard to morbidity, mortality and long-term outcome are unsatisfactory. Today the surgical standard is careful digital necrosectomy, which must be followed by either postoperatve drainage, repeated open lavage or continuous closed lavage. In experienced hands, all three procedures provide similar results with regard to morbidity and mortality. PMID- 10789045 TI - [Operational costs and control of performance in surgical clinics between marketing and planning economics. Risk or perhaps quadrature of the circle]. AB - Surgical hospitals can be seen as operational or even industrial production systems. Doctors have a major impact on both medical performance and costs. For active participation in the management process, knowledge of industrial controlling mechanisms is required. German hospitals currently receive no procedure-related financial revenues, such as prices or tariffs for defined medical treatment activities. Maximum clinical revenues are, furthermore, limited by principles of planned economy and can be increased only slightly by greater medical performance. Costs are the only target that can be autonomously influenced by the management. Operative controlling in hospitals aims at horizontal and vertical coordination of subunits and decentralization of process regulations. Hospital medical performance is not clearly defined, its quantitative measurement very problematic. Process-orientated clinical activities are not taken into account. A high percentage of hospital costs are fixed and can be influenced only by major structural interventions in the long term. Variable costs are primarily dependent on the quantity of clinical activities, but also heavily influenced by patient structure (comorbidity and risk profile). The various forms of industrial cost calculations, such as internal budgeting, internal markets or flexible plan-cost balancing, cannot be directly applied in hospital management. Based on these analyses, current operational concepts and strategic trends are listed to describe cost-management options in hospitals with focus on the German health reforms. PMID- 10789046 TI - [Improved detection of gastrointestinal bleeding sites with 99mTc market autologous erythrocytes and continuous dynamic scintigraphy with cine-mode display]. AB - BACKGROUND: Scintigraphic studies of gastrointestinal bleeding are usually performed in static acquisition mode and without continuous imaging over a longer period. The aim of this study was to evaluate the diagnostic accuracy of continuous dynamic 99mTc red blood cell scintigraphy (BQS) and cine-mode display in the assessment of unexplained gastrointestinal bleeding. PATIENTS AND METHODS: We performed BQS in 40 patients (mean age 57 years) with gastrointestinal bleeding from an undetermined source. All these patients had negative findings of previous examinations. Blood transfusions were required in 16 patients. Continuous dynamic scintigraphic acquisition was performed up to 6 h post injection (p.i.). The scintigraphic data were reviewed without knowledge of the patient's final diagnosis. For this purpose we used a cine-mode display. RESULTS: Dynamic 99mTc red blood cell scintigraphy correctly identified the site of active bleeding in 22 of 23 patients with positive scintigraphic findings. The sensitivity of the scan was 95%, the specificity 94%, the positive and negative predictive values 95% and 94%, respectively. Forty-three percent of the scans became positive within 1.5 h and 82% within 5 h. Positive scans were more likely in patients who required blood transfusions than in patients without transfusion requirement (16 vs 7 patients), and the bleeding site was identified earlier in the former group (mean 2.83 vs 5.28 h p.i.). The scintigraphic identification of the bleeding lesions enabled the performance of a more limited surgical approach in the colon as well as in the distal small intestine, while exact scintigraphic localization was not possible if the bleeding site was located in the proximal small intestine. DISCUSSION: Continuous dynamic 99mTc red blood cell scintigraphy interpreted by cine-mode display is highly accurate in the identification of a bleeding site. When performed correctly, scintigraphy enables usually segmental resection of these lesions. PMID- 10789047 TI - [Primary varicose veins--changes in the venous wall and elastic behavior]. AB - The aetiology of varicosis is still unclear. Investigation of changes in normal veins which result in the formation of varicosis led to examination of elastic properties and histomorphological alterations in the region of the extracellular matrix in the vessel wall of the long saphenous vein in normal and varicose veins. The rigidity of varicose veins was higher than that of normal veins; there was no difference between the varicose vein above a competent valve and the "normal" vein below this competent valve. The matrix proteins demonstrated an increase in total fibres, whereas the elastic fibres were reduced and fragmented, explained the rigidity of varices. These results suggest that the role of the venous valves in the development of varicose veins is secondary to changes of the vein wall. PMID- 10789048 TI - [Is the Bishop-Koop anastomosis in treatment of neonatal ileus still current?]. AB - Fourty-three cases of ileus in newborns are presented. Twenty-seven newborns received a Bishop-Koop anastomosis. In 19 cases, the Bishop-Koop anastomosis was performed primarily and in 8 cases as a second intervention. We consider the Bishop-Koop anastomosis to be a safer procedure than primary end-to-end or end-to side anastomosis. Only one anastomotic leak occurred in our patients. We prefer the Bishop-Koop anastomosis not only in cases of meconium ileus, but also in other types of intestinal atresia and stenosis, especially for the management of greatly different intestinal diameters. In our experience, this method is also suitable for re-anastomosing a double-barrel anastomosis. The Bishop-Koop procedure minimizes the risks of primary anastomosis without enterostoma, and later extraperitoneal closure of the stoma is easy and safe. PMID- 10789049 TI - [Therapeutic options in treatment of heel defects. Presentation of an algorithm for therapeutic strategy]. AB - INTRODUCTION: The treatment of soft tissue defects of the foot and ankle region remains a challenge, because special anatomical and physiological properties have to be considered. PATIENTS AND METHODS: The data of 52 patients, who were treated between 1994 and 1998, were analyzed. 48% were posttraumatic defects and were located in the weight bearing zone in 37%. RESULTS: Fifty-nine procedures were used, including skin transplantation in 16 patients, local flaps in 3 cases, regional flaps in 14 cases, and microvascular procedures in 17 cases. Amputation was required in 4 patients. The overall success rate was 92%. Complications occurred in 18 patients. CONCLUSIONS: Using these data and evaluation of the actual literature lead to the development of an algorithm for the treatment of defects in this area. The modern armamentarium of plastic surgery offers the possible different treatment modalities. PMID- 10789050 TI - [Osteoid osteoma--a diagnostic and therapeutic problem?]. AB - INTRODUCTION: The aim of the following article is to summarize our experience in the treatment of osteiod osteomas, with special emphasis on diagnostic and therapeutic pitfalls. PATIENTS AND METHODS: A consecutive series of 14 patients with osteoid osteomas was treated surgically between 1985 and 1996. All tumors but one were located in the lower limb. The main symptom was pain, being worse at night and being responsive to oral salicylates (10/14 patients). As reported in other studies, the duration of symptoms was unacceptably long (mean 24 months). The classical pathognomonic symptoms were misinterpreted in many cases, leading to frustrating conservative and even operative therapies. RESULTS: Open biopsy prior to surgical excision is not indicated because of the typical clinical and roentgenographic imaging of these lesions. Surgical excision of the nidus is the treatment of choice and gives immediate pain relief. If the characteristic morphology is not evident in plain roentgenograms, conventional tomograms, radionuclide scans and computerized tomography are reliable tools. All patients were free of disease at a mean of 6.6 years after operation. CONCLUSION: In symptomatic patients with osteoid osteomas the excision of the nidus is the established diagnostic/therapeutic modality. Minimally invasive procedures seem to be alternatives to classical surgery. PMID- 10789051 TI - [Orthograde Kirschner wire osteosynthesis. Experiences with intramedullary fixation of the distal metacarpal V fracture]. AB - Dislocated metacarpal V fractures, fixed by percutaneous Kirschner wires or other implants near the metacarpophalangeal joint, often show impaired movement despite excellent radiologic findings. We therefore chose an alternative method using intramedullary Kirschner wire osteosynthesis according to Foucher. Twenty-five patients were operated on in a 3-year period and movement of the fingers and complications were examined. Six weeks after operation all fractures were consolidated in anatomical reduction. One patient suffering from an early stage of Sudeck's dystrophy was healed, and no other complications appeared. Three months after treatment all patients achieved free movement of the fingers with anatomical alignment. We therefore recommend intramedullary Kirschner wire osteosynthesis in distal metacarpal V fractures. PMID- 10789052 TI - [Duodenal diverticulum--an unusual manifestation of recurrent retroperitoneal liposarcoma]. AB - The treatment of choice in retroperitoneal liposarcoma is curative R0 resection. However, there are high rates of reoperation due to recurrence. Our present case report shows difficulties in detecting recurrence due to atypical configuration. Eventually a huge diverticulum of the duodenum combined with leiomyoma in its neck area was histopathologically identified as a recurrent liposarcoma. PMID- 10789054 TI - [Development of large vein surgery in Europe]. AB - Surgery of the great veins inevitably began with the surgical treatment of injuries, often involving the femoral vein. Because of the famous case presented by Roux in 1813, the prevailing opinion until almost the end of the nineteenth century was that ligation of the vein made death inevitable, but that the only way to control severe hemorrhaging from a vein was to ligate the femoral artery. Zaufal's principle (1880) consisted in the ligation and resection of great veins of the body and limbs in order to prevent sepsis in suppurative processes. This surgical method was still being used in the first half of the twentieth century in patients with perforated appendicitis or puerperal fever. In the limb, the congestion induced in the vein had a positive effect on the healing process. The surgical treatment of leg and pelvic deep vein thrombosis was initially (1931) aimed at achieving decompression in compartment syndromes. The first thrombectomy was performed in 1937, but several operations with a successful outcome were reported at the 61st annual meeting of the German Society of Surgery in the following year. PMID- 10789053 TI - [Primary liposarcoma of the stomach wall--a rare mesenchymal tumor]. AB - INTRODUCTION: Primary liposarcoma of the stomach wall is rare. Only eight cases have been described so far. MATERIAL AND METHOD: Here we report the ninth case, occurring in a 74-year-old woman who presented with weight loss and a therapy resistant ulcer of the stomach wall. RESULTS: Pre- and perioperative findings suggested a benign lipoma of the stomach wall. The patient was treated with subtotal gastrectomy. On microscopic examination the tumor showed features of a benign lipoma but for a distinctive capillary net. Immunohistochemically the S 100 reaction was positive. Less than 1% of Ki67-positive cells could be found, thus suggesting a highly differentiated primary liposarcoma of the stomach wall. CONCLUSION: In situations where the benign or malignant nature of a submucosal lesion cannot be diagnosed with certainty a mesenchymal tumor of the stomach wall has to be included in the differential diagnosis. Here the indication for complete surgical excision and histological workup has to be set widely. PMID- 10789055 TI - [Therapy concepts in femoral neck fractures. 2]. PMID- 10789056 TI - [Quality management--needs and reality. Opening address at the 24th Interdisciplinary Forum "Progression and Continuing Education in Medicine" by the Federal Physicians Group, 12-15 January 2000]. PMID- 10789057 TI - [Structures for solidarity in public health insurance. Public health insurance must strive for acceptance in public health policy]. PMID- 10789058 TI - [Legal occupational problems of hepatitis C/AIDS infection in physicians]. PMID- 10789059 TI - Some physicochemical properties of mefenamic acid. AB - Mefenamic acid is a problematic drug in granulation, tableting, and dissolution due to its poor solubility, hydrophobicity, and tendency to stick to surfaces. In most cases, the specifications of a drug by the pharmacopoeia include identification and purity, but they do not describe the physicochemical drug properties precisely. To characterize the mefenamic acid particle size, surface area measurements, X-ray pattern, differential scanning calorimetry (DSC), wettability, crystal habit, and compression behavior of different batches from two manufacturers were investigated. Due to larger particle size and better wettability, mefenamic acid of Il Yang type was easier to handle in a granulation process. The compression behavior of both types was nearly the same, although particle size, crystal habit, and wettability were very different. PMID- 10789060 TI - Factors influencing capping and cracking of mefenamic acid tablets. AB - The tendency of capping and longitudinal cracks of mefenamic acid tablets was evaluated in relation to the amount of the binder, the influence of the granulation technique, and the relative humidity of the granules. Tablets made from fluidized bed granules using methylcellulose in the granulating liquid showed significantly lower capping and longitudinal cracks than tablets from conventional granules prepared by wet granulation using methylcellulose as a dry binder. PMID- 10789061 TI - Investigation of the mechanical properties of two polyvinyl alcohols and their influence on drug release. AB - The aim of this work was the investigation of the elastic behavior and the internal stress of two different polyvinyl alcohols (PVAs) and how their swelling behavior and drug release are influenced through internal stress. We used thermal mechanical analysis (TMA 202/1/F Netzsch-Germany) to make a qualitative evaluation of the internal stress and the elastic behavior of PVA 55/12 and 55/03. These two PVAs have the same molecular weight and different acetyl contents. The tablets were compressed with an instrumented eccentric tablet machine. Tablets from different PVAs showed different elastic behavior. The measured internal stress of PVA 55/12 was higher than that of PVA 55/03. By increasing the compaction force, the internal stress increased. The internal stress decreased as the humidity increased for both kinds of PVA. The expansion behavior in the diametrical and axial direction was significantly different due to the predominant orientation of the PVA crystals. PVA tablets that were thermal analyzed showed different swelling behavior from those that were not thermal analyzed. This can be caused by the released internal stress of thermal-analyzed tablets. The compaction behaviors of both PVAs were also investigated using different methods. PVA 55/03 had better compactibility and compressibility behavior. The drug release behavior was also investigated and showed that the drug release rate of PVA 55/12 was higher than that of PVA 55/03. This can be due to the different solubilities and elastic behaviors of the PVAs. TMA can be used to predict and evaluate elastic behavior and internal stress. Humidity can influence the internal stress of the tablet. The measured swelling of PVA was a combination of real swelling and the released internal stress. TMA also allowed prediction of the binding mechanism in tablets. PMID- 10789062 TI - Aerosol-OT microemulsions as transdermal carriers of tetracaine hydrochloride. AB - The aerosol-OT (AOT)/water/isopropyl myristate microemulsion was investigated as a carrier in transdermal drug delivery of tetracaine hydrochloride. The study included in vivo analgesic studies on rats and histopathological, irritation, and oxidative stress measurements on mice. The tetracaine hydrochloride encapsulated in AOT/water/isopropyl myristate showed an eightfold enhancement in the analgesic response of drug compared to the aqueous solution of the drug as measured by the tail-flick method. The analgesic response of tetracaine hydrochloride, however, highly depended on the concentration of AOT and water of the microemulsion. The preliminary histopathological, irritation, and oxidative stress studies showed that AOT/water/isopropyl myristate microemulsion system is a safe transdermal carrier of tetracaine hydrochloride with a concentration of AOT in isopropyl myristate up to 21:79 w/w. PMID- 10789063 TI - Formulation and development of tablets based on Ludipress and scale-up from laboratory to production scale. AB - In spite of the wealth of experience available in the pharmaceutical industry, tablet formulations are still largely developed on an empirical basis, and the scale-up from laboratory to production is a time-consuming and costly process. Using Ludipress greatly simplifies formulation development and the manufacturing process because only the active ingredient Ludipress and a lubricant need to be mixed briefly before being compressed into tablets. The studies described here were designed to investigate the scale-up of Ludipress-based formulations from laboratory to production scale, and to predict changes in tablet properties due to changes in format, compaction pressure, and the use of different tablet presses. It was found that the tensile strength of tablets made of Ludipress increased linearly with compaction pressures up to 300 MPa. It was also independent of the geometry of the tablets (diameter, thickness, shape). It is therefore possible to give an equation with which the compaction pressure required to achieve a given hardness can be calculated for a given tablet form. The equation has to be modified slightly to convert from a single-punch press to a rotary tableting machine. Tablets produced in the rotary machine at the same pressure have a slightly higher tensile strength. The rate of increase in pressure, and therefore the throughput, has no effect on the tensile strength of Ludipress tablets. It is thought that a certain minimum dwell time is responsible for this difference. The production of tablets based on Ludipress can be scaled up from one rotary press to another without problem if the powder mixtures are prepared with the same mixing energy. The tensile strength curve determined for tablets made with Ludipress alone can also be applied to tablets with a small quantity (< 10%) of an active ingredient. PMID- 10789064 TI - Preparation and in vitro evaluation of self-microemulsifying drug delivery systems containing idebenone. AB - A new self-microemulsifying drug delivery system (SMEDDS) was developed to increase the dissolution rate, solubility, and, ultimately, bioavailability of a poorly water soluble drug, idebenone. Pseudoternary phase diagrams were used to evaluate the self-microemulsification existence area, and the release rate of idebenone was investigated. The mixtures consisting of Labrafac hydro or Labrafil 2609 (HLB values > 4) with the surfactant (Labrasol containing 80% Transcutol) and cosurfactant (Plurol oleique WL 1173) were found to be optimum formulations. Using the SMEDDS formulations of 5% to 20% of Labrafac hydro or Labrafil 2609 in combination with the surfactant/cosurfactant mixing ratio of 3, the microemulsion existence field was wider compared to the other SMEDDS formulations due to high affinity for the continuous phase. The in vitro dissolution rate of idebenone from SMEDDS was more than twofold faster compared with that of tablets. The developed SMEDDS formulation can be used as a possible alternative to traditional oral formulations of idebenone to improve its bioavailability. PMID- 10789065 TI - Characterization of zopiclone crystal forms found among generic raw materials. AB - This paper deals with the occurrence of polymorphs and pseudopolymorphs and their effect on the solid-state properties of zopiclone, a poorly water soluble nondiazepine sedative and hypnotic drug. X-ray powder diffraction (XRPD), infrared spectroscopy (IR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), particle size analysis, dissolution studies, and solubility determinations were used to characterize the zopiclone raw materials. An anhydrated form, a dihydrated form, and a mixture of these two crystal forms were found and characterized among the zopiclone powders. PMID- 10789066 TI - Comparing HPLC and UV spectrophotometric analysis methods for determining the stability of sorbic acid in nonionic creams containing lactic acid. AB - This paper describes a comparison between ultraviolet (UV) spectrophotometric and high-performance liquid chromatographic (HPLC) methods of analysis for the determination of sorbic acid in nonionic creams containing lactic acid. Sorbic acid is an antimycotic agent and is used as a preservative in pharmaceuticals, cosmetics, and food products. UV spectrophotometric analysis was done by calculating the concentration of remaining sorbic acid from the absorbance values and the molar extinction coefficient EM258 = 24,080. A decrease in absorbance at 258 nm was accompanied by a simultaneous increase in total carbonyls and monoaldehyde content and the appearance of a very weak absorption maximum between 215 and 225 nm. HPLC analysis was done with a Hypersil BDS C8 column with detection at 254 nm and employing a mobile phase consisting of a mixture of buffer and methanol (7:3 v/v) at a pH of 2.25. The buffer consisted of 0.85% H2SO4 in 17.5 mM KH2PO4. The validation results, together with statistical treatment of the data, demonstrated the reliability of both procedures. A drawback of the UV methods was, however, its lack of adequate measurement of sorbic acid stability at higher temperatures. For these assays, the HPLC method was found to be adequate, and it should therefore be used to obtain accurate stability data for sorbic acid in creams. PMID- 10789067 TI - A pH-dependent colon-targeted oral drug delivery system using methacrylic acid copolymers. II. Manipulation of drug release using Eudragit L100 and Eudragit S100 combinations. AB - Tablets containing mesalazine as a model drug were coated using various combinations of two methacrylic acid copolymers, (Eudragit L100 and Eudragit S100) by spraying from aqueous systems. The Eudragit L100-Eudragit S100 (w/w) combinations studied were 1:0, 4:1, 3:2, 1:1, 2:3, 1:4, 1:5, and 0:1. The coated tablets were tested in vitro for their suitability for pH-dependent colon targeted oral drug delivery. The dissolution profiles of the drug obtained from the studied tablets demonstrate that the release of the drug could be manipulated by changing the Eudragit L100-Eudragit S100 ratios in the combinations within the pH range between 6.0 and 7.0 in which the individual polymers are soluble, and a coating formulation consisting of a combination of the two polymers can overcome the issue of high gastrointestinal (GI) pH variability among individuals. The results also demonstrate the feasibility of using aqueous dispersions of Eudragit L100-Eudragit S100 combinations for coating tablets for colon-targeted delivery of drugs, and that the formulation can be adjusted to deliver drug(s) at any other desirable site of the intestinal region of the GI tract in which pH of the fluid is within the range 6.0 to 7.0. For colon-targeted delivery of drugs, the proposed combination system is superior to tablets coated with either Eudragit L100 or Eudragit S100 alone. PMID- 10789068 TI - A temperature study on critical micellization concentration of the novel platelet activating factor receptor antagonist E5880 in water by electric conductivity measurements. AB - To clarify the behavior of the novel platelet-activating factor (PAF) receptor antagonist E5880 in aqueous solution, electric conductivity was measured at different temperatures (every 5 degrees C), ranging from 15 degrees C to 50 degrees C. Critical micellization concentration (CMC) of E5880 was dependent on the temperature; at 30 degrees C, the CMC value was smallest (0.143 mM). Below that temperature, the enthalpy for formation of the micelle (delta Hm0) was positive, and the formation of micelles was endothermic; above that temperature, delta Hm0 was negative, and the formation of micelles was exothermic. PMID- 10789069 TI - Administration of human chorionic gonadotropin with a controlled-release function to immature rats for application in male infertility therapy. AB - Biodegradable poly(DL-lactic acid) (PLA) devices with controlled release of human chorionic gonadotropin (hCG) were fabricated in a 2-mm cylinder form by the meltpressing technique. The devices fabricated were inserted subcutaneously in the backs of immature rats. It was found that the serum level of testosterone (T) remained constant at approximately 0.7 ng/ml for an experimental period of 14 days, resulting in greater Leydig cell production of testosterone. This was also suggested from the results of immunohistochemical observation of the testis and weight changes of prostates, such as the ventral prostates and seminal vesicle. PMID- 10789070 TI - Enhancing effects of fatty acids on piroxicam permeation through rat skins. AB - To increase the skin permeation of piroxicam from the Poloxamer 407 gel, fatty acid was added as a penetration enhancer to the Poloxamer 407 gel containing 1% piroxicam. The enhancing effects of the enhancer on the skin permeation of piroxicam were evaluated using Franz diffusion cells fitted with intact excised rat skins. To elucidate the modes of the action of enhancers, thermal analysis and histological examinations were conducted. Among fatty acids tested, linoleic acid showed the highest enhancing effects, with an enhancement factor (EF) of 1.76. From the thermal analysis results, fatty acids have fluidizing effects on the stratum corneum. The skin pretreated with the Poloxamer 407 gels containing piroxicam including linoleic acid showed a loosely layered stratum corneum and wide intercellular space. PMID- 10789071 TI - Improved bioavailability of a micronized megestrol acetate tablet formulation in humans. AB - Megestrol acetate, a progestogen widely used in the palliative treatment of endometrial carcinoma and breast cancer, is currently administered orally as a solid dosage form. Bioavailability of the drug following oral administration is closely related to the effectiveness and safety profile of the drug in formulation. Improved immediate-release formulations should allow improved drug delivery into the systemic circulation and, at the end, to the site of action. The micronization of drugs is one of the technological procedures to achieve such a purpose. This paper reports the design and results obtained in an in vivo study of the bioavailability of a micronized megestrol acetate tablet formulation compared to a conventional form. A significant increase in the drug bioavailability was observed, in either the rate or the extent of absorption. In vitro dissolution data of the two study formulations reflected the in vivo findings. PMID- 10789072 TI - The contributions of erosion, swelling, and porosity to theophylline release kinetics from Cissus populnea polymer matrices. AB - The variable factors of erosion rate, swelling rate, and porosity were used in studying the release patterns of theophylline from our Cissus populnea polymer (CPP) matrices under the different factor combinations given by a simple 2n factorial experimental design. The zero-order slopes and correlation coefficients representing release rate and linearity, respectively, as obtained from both the nonsteady state and steady state were statistically treated. It appears that the nonsteady-state analysis is more suitable for studying the effects of individual factors, while the steady-state analysis appears more suitable for studying interaction effects of the factors. The study also showed that erosion is the main mechanism by which theophylline is released from the matrices, while swelling is responsible for maintaining linearity in the zero-order release curves. A careful examination of the statistical results shows some functional relationships between the factors, which should be considered in designing more detailed factorial experiments to enable the establishment of equation models for predicting the release profile of theophylline from our CPP matrices under any given dissolution condition. PMID- 10789073 TI - Formulation optimization of controlled-release pellets of metoclopramide hydrochloride using dissolution fit factor approach. AB - The purpose of this study was to optimize the formulation variables for the preparation of ethyl cellulose-coated nonpareils loaded with metoclopramide hydrochloride (MCL). The approach to evaluate the effectiveness of formulation parameters was monitored by release rate testing using dissolution fit factors as a tool. The content of ethyl cellulose used in the formulation was based on the drug-loaded weight. The interrelationship of each developed formulation and the reference formulation Gastro-Timelets and their respective dissolution curves were evaluated using Moore's equation: [equation: see text]. The relationship between the ethyl cellulose content in the formulation and the dissolution fit factor f2 can be described as the following regression equation: Y = -0.054X2 + 3.347X - 1.915 (r2 = 0.99). The optimum ethyl cellulose content obtained from the equation was 30.8%. The type and content of plasticizer used in the formulation to achieve the greatest f2 were determined to be Myvacet 9-40 at the concentration of 25%. Results indicated that using the release rate testing approach with the dissolution fit factor as a tool could provide valuable information for formulation optimization. PMID- 10789074 TI - [Vulvodynia]. AB - Vulvodynia is a complex syndrome of chronic vulvar pain. It is divided into several subtypes: 1. cyclic vulvovaginitis (pain occurs after coitus), 2. vulvar vestibulitis syndrome (pain mainly with intercourse), 3. dysaesthetic vulvodynia (psychosomatic; diagnosis of exclusion), 4. vulvar dermatoses (e.g. pemphigus vulgaris, contact dermatitis). Additional causes have been described in single cases. Though vulvodynia is often accompanied by psychological distress, somatic causes have to be considered in each case. PMID- 10789075 TI - [The neodymium YAG laser: its application in dermatology]. AB - The neodymium:yttrium-aluminium-garnet-(Nd:YAG) laser is one of the most versatile and interesting lasers in dermatological laser medicine. Almost no other laser has such a wide spectrum of applications. Depending on wave length (1064 nm, frequency-doubled 532 nm) and mode (continuous, Q-switched, long pulsed), benign pigmented lesions (tattoos, traumatic tattoos, naevus of Ota, cafe-au-lait-macules, lentigines) and vascular lesions (hemangiomas, portwine stains, essential teleangiectasias, angiomas) can be treated. Additionally this laser has been occasionally used to remove Kaposi sarcomas and epithelial skin tumors. PMID- 10789076 TI - [Lipodystrophy syndrome in HIV infection]. AB - Since the availability of protease inhibitors in 1997, there has been a great change in antiretroviral therapy but also new long term side effects have emerged, mainly metabolic changes such as hypertriglyceridemia, hypercholesteremia and hyperglycemia. Besides, fat redistribution has been observed. Fat wastes in the face and limbs but accumulates in the adipose tissue of the dorsocervical and abdominal region and the breasts. The mechanism of these changes remains unclear. For therapy a protease inhibitor free therapy or lipid lowering drugs may be tried. 29 of our 224 patients developed lipodystrophy. 27 of these patients had been treated with a protease inhibitor (17 patients with indinavir); 2 of the patients had never received protease inhibitors. PMID- 10789077 TI - [Topical administration of capsaicin in dermatology for treatment of itching and pain]. AB - BACKGROUND AND OBJECTIVES: Treatment of pruritus as observed in many dermatological and internal diseases may be very often disappointing. There are many reports describing the topical use of capsaicin, the pungent agent in red pepper. Long term administration of capsaicin depletes neuropeptides in unmyelinated, polymodal C-type and small myelinated A delta-type cutaneous nerves that conduct pruritus and pain. PATIENTS/METHODS: The aim of this study was to evaluate the efficacy, safety and practicability of capsaicin in the treatment of pruritus and pain of different origin in a total of 40 patients. RESULTS: Capsaicin suppressed completely itch in all patients. CONCLUSIONS: In addition to its excellent antipruritic and analgetic effect, capsaicin significantly contributed to healing of some dermatoses, such as prurigo nodularis and psoriasis. PMID- 10789078 TI - [Use of interferon-alpha (IFN-alpha) in the treatment of keratoacanthoma]. AB - BACKGROUND AND OBJECTIVE: Keratoacanthoma are benign epithelial tumors. The aim of this study was to determine the effects of interferon alpha on keratoacanthoma. PATIENTS/METHODS: Six patients presented with histologically proven keratoacanthoma which were large or otherwise difficult to excise. Intra and perilesional interferon-alpha treatment with 3 million IU weekly was started. RESULTS: In 6-15 weeks all tumors were cured. CONCLUSIONS: Treatment with intralesional interferon-alpha offers a new possibility in cases of large keratoacanthomas, or those which are not surgically excisable. PMID- 10789079 TI - [Implicit personality theories in dermatology. An empirical study on the image that physicians have of patients of diverse dermatologic diagnosis groups]. AB - BACKGROUND AND OBJECTIVE: Studies about the personality of dermatological patients are numerous. The results are inconsistent and peculiarities of the personality are rarely confirmed. Contrary to the usual procedure, this study asks for the impressions physicians have about their patients. PATIENTS/METHODS: To elicit associations dependent on a dermatological diagnosis, 27 physicians filled in a three-dimensional differential asking for their impressions about patients with atopic dermatitis, leg ulcer, malignant melanoma, or psoriasis vulgaris in order to quantify these impressions. RESULTS: The dimension valence (good-bad) did not differ between the groups. The dimensions of excitation (active-passive) and potency (strong-weak) showed the biggest differences between patients with atopic dermatitis and ulcus cruris. The latter get significantly lower values on both dimensions. The correlations between the impression and the duration of employment in dermatology were low. CONCLUSIONS: In our opinion it is necessary to raise the impression of medical personal about their patients because these impressions affect the interaction of the personal with the patient. PMID- 10789081 TI - [Radiation-induced acne]. AB - Radiation-induced acne is a rare, clinically and pathogenetically ill-defined acneiform dermatosis with special features that may occur in irradiated skin areas especially after high doses of deeply penetrating radiation. We report on a patient with an oropharyngeal carcinoma who developed severe radiation-induced acne including comedones and cysts as well as few inflammatory papules and pustules in a skin area irradiated with up to 63 gray of a 6 MeV photon beam. Acnegenic drugs may precipitate the disease; our patient was on longterm therapy with carbamazepine whose acnegenic potency is less well documented than that of testosterone or glucocorticoids. Treatment of radiation-induced acne is comedolytic; topical retinoids are especially valuable. PMID- 10789080 TI - [Eosinophilic cellulitis (Wells syndrome)]. AB - Eosinophilic cellulitis (Wells' syndrome) is a rare disorder characterized clinically by recurrent erythematous plaques resembling cellulitis and histologically by a dermal infiltrate of lymphocytes, eosinophils and eosinophil debris between collagen bundles, forming flame figures in typical cases. A 71 year-old woman with Wells' syndrome with blood and bone marrow eosinophilia showed a good response to dapsone. The level of eosinophil cationic protein (ECP) in serum was elevated. Immunophenotyping of peripheral T cells revealed an increased proportion of CD3+CD4+T cells. The patients' cultured peripheral lymphocytes spontaneously released significant amounts of interleukin 5 (IL-5), but not interleukin 4 (IL-4) or interferon gamma (IFN gamma). These findings suggest that activated T cells may be involved in the pathogenesis of blood and tissue eosinophilia in this patient. PMID- 10789082 TI - [Congenital pachyonychia type II (Jackson-Lawler syndrome)]. AB - Pachyonychia congenita (PC) is a rare ectodermal dysplasia with variable expression. The condition is usually inherited as an autosomal dominant trait. Several classifications of PC have been proposed. Feinstein and colleagues suggested four clinical types of PC. Type II, the Jackson-Lawler-Syndrome, is characterized by multiple epidermal cysts, palmoplantar bullae and hyperhidrosis as well as natal teeth in addition to the main findings of pachyonychia, palmoplantar hyperkeratosis and follicular keratosis. We report two patients (father and son) with Jackson-Lawler-Syndrome and describe in detail pathogenesis, diagnostic criteria and treatment approaches as well as the different classifications of pachyonychia congenita. PMID- 10789083 TI - [Follicular dyskeratosis: successful treatment with local retinoid]. AB - Therapy of Darier disease is rarely satisfactory. Oral retinoids can improve the skin lesions. Because of previous positive results with topical retinoids, which were often not practical because of marked skin irritation, two patients were treated with the new, topical acetylene-retinoid tazarotene in a 0.01% gel preparation. In both patients skin cleared within 2 to 4 weeks. After stopping the medication patients did not develop any new skin lesions for 12 months. In a third patient, healing with tazarotene gel 0.025% was faster than with a topical preparation containing 10% urea. Tazarotene is an important addition to the therapeutic possibilities in Darier disease. PMID- 10789084 TI - [Prevention of pyoderma gangrenosum]. PMID- 10789085 TI - [Plica polonica in the 21st century]. AB - We describe a young man presenting with dreadlocks. There are remarkable similarities with the so called plica polonica, that historically had been treated with long courses of mercury. Apparently very important in the 18th century, the interest for this hair-disorder appears to be lost in specialized medical literature. In contrast dreadlocks, a recent hairstyle are frequently encountered. Lack of other sources various websites provide dermatologists with answers to questions regarding complications. Fortunately a simply haircut is today treatment enough. PMID- 10789086 TI - [Irritant contact dermatitis]. PMID- 10789088 TI - [Arterial hypertension and heart insufficiency]. PMID- 10789087 TI - [Arrhythmias in heart insufficiency]. PMID- 10789089 TI - [Biventricular stimulation in heart insufficiency]. PMID- 10789090 TI - [Exercise tolerance in patients with heart insufficiency. Exercise in heart insufficiency?]. PMID- 10789091 TI - [Anticoagulation in heart insufficiency]. PMID- 10789092 TI - [Perioperative risk and preoperative preparation of the cardiac patient for noncardiac surgery]. PMID- 10789093 TI - [Cerebrospinal fluid analysis]. PMID- 10789094 TI - [Pancytopenia, hepatosplenomegaly and fever in a 49-year old patient]. PMID- 10789095 TI - [Thirty-year old woman with severe iron deficiency anemia and weight loss of unknown etiology]. PMID- 10789096 TI - [Thyroid hormone therapy]. PMID- 10789097 TI - Biomechanics of the first ray. Part II: Metatarsus primus varus as a cause of hypermobility. A three-dimensional kinematic analysis in a cadaver model. AB - Variation in functional stability of the first metatarsocuneiform joint was analyzed between transverse plane deviated (adducted) and corrected first metatarsal positions in a closed kinetic chain model. Six fresh frozen cadaver specimens with intact ankles and feet were fitted with a custom fabricated titanium metatarsal jig, which allowed for manipulation of the first metatarsal in the transverse plane. Specimens were mounted into a custom-made acrylic load frame and axially loaded to 400 N. Radiowave three-dimensional tracking transducers were attached to the following osseous segments: first metatarsal head and base, medial cuneiform, and second metatarsal. A dorsally directed load was applied to the first metatarsal segment and resultant movements were measured. Repeated testing was performed on a transverse deviated and corrected first metatarsal positions with the hallux plantargrade and maximally dorsiflexed to engage the windlass mechanism. With the windlass mechanism engaged and first metatarsal corrected, a 26% increase in first ray plantarflexion occurred from a deviated to a corrected first metatarsal position (p < or = .05). This suggests that the windlass mechanism is more efficient when the first metatarsal, sesamoid apparatus, and hallux position are properly aligned with the orientation of the plantar aponeurosis. Clinically, this may explain the correlation of first ray hypermobility with the progression of bunion severity. Our study validates the earlier work of Hicks and adds additional insight into the functional stability in the medial column of the foot. PMID- 10789098 TI - The effects on subtalar contact and pressure following talonavicular and midtarsal joint arthrodesis. AB - Many clinical studies have demonstrated the effectiveness of both isolated talonavicular and complete midtarsal joint arthrodesis as an alternative to triple arthrodesis. However, in many cases, controversy exists as to which procedure to utilize. Evidence of degenerative radiographic changes and stiffness of the subtalar joint have been reported postoperatively. A cadaveric study at two different loading values, utilizing low-range pressure film transducers and digital scanning, was performed to quantify articular contact effects on the subtalar joint following isolated talonavicular joint arthrodesis and complete midtarsal joint arthrodesis as compared to the intact specimen. Statistically significant differences were found at p < .05 in this study regarding maximum contact pressure and in location of the applied pressures. Results of this study suggest complete midtarsal joint arthrodesis may be favored over isolated talonavicular joint arthrodesis, especially in the setting of a flatfoot deformity. PMID- 10789099 TI - Relative stability of tension band versus two-cortex screw fixation for treating fifth metatarsal base avulsion fractures. AB - This study assesses the strength of fixating avulsion fractures of the fifth metatarsal base with a 4.0-mm partially threaded cancellous screw crossing two cortices as compared to tension banding. Our data showed statistically significant fixation strength improvement over tension banding for avulsion fractures (p < 0.02) in both polystyrene foam models and fresh, nonpreserved frozen cadaveric samples. In cadavers, the screw fixations were able to withstand more than three times the load sustained by the tension band fixations. The study utilized the Instron 8500 tensiometer to apply physiologic loads to test the constructs until failure. The displacement and load data at failure show the limitations of both fixations. By increasing the load resistance while maintaining compression, the bicortical cancellous screw fixation created greater stability at the avulsion fracture of the fifth metatarsal base as compared to tension band stabilization. PMID- 10789100 TI - Results of treatment of 22 navicular stress fractures and a new proposed radiographic classification system. AB - Twenty-two navicular stress fractures sustained during athletic activity were retrospectively reviewed for return to activity time and the appearance of fracture pattern on computerized tomography. There were 10 females and nine males, with the average patient age being 27.2 years. Three patients sustained bilateral injuries at separate times. Average follow-up was 36.5 months. Nine patients underwent open reduction, internal fixation (some with bone grafting); this group's average return to activity (RTA) was 3.1 +/- 1.2 months (range, 1.5 5 months). Thirteen patients treated conservatively had an average return to activity of 4.3 +/- 2.8 months (range, 2-13 months). The difference between the two groups' RTA was significant (p = .02). Eleven patients utilized pulsed electromagnetic fields (PEMF) and had an average RTA of 4.2 +/- 3.4 months, 27.3% of those patients with PEMF also had surgery. Two conservatively treated fractures that took 5 and 8 months to RTA, respectively, re-fractured during the treatment process. Retrospective review showed CT fracture patterns in the frontal plane that were classified as: dorsal cortical break (type I), fracture propagation into the navicular body (type II), and fracture propagation into another cortex (type III). This is a proposed classification system. It includes modifiers "A" (avascular necrosis of a portion of the navicular); "C" (cystic changes of the fracture), and "S" (sclerosis of the margins of the fracture), the latter of which was most common in our series, particularly in continually symptomatic patients. Type I fractures were more likely to receive conservative treatment (p = .02) and type III fractures took significantly longer to heal than types I and II (p values .001 and .01, respectively). Type I and II injuries had an average RTA of 3.0 and 3.6 months, respectively. Type III injuries had an average RTA of 6.8 months. Based on our findings, we recommend surgery for patients with these modifiers, particularly with type II and III injuries. Conservative treatment may be prolonged, and requires at least 6 weeks of nonweightbearing in a below-knee cast/boot to be successful. PMID- 10789101 TI - Proximal shortening lesser metatarsal osteotomy: a mathematical-geometric basis. AB - Reconstruction and restoration of the lesser metatarsal parabola after an iatrogenic complication of a lesser metatarsal osteotomy provides a difficult surgical dilemma for the foot and ankle surgeon. This study's purpose was to determine if a formula could be developed, through a geometric and mathematical basis, for the proximal shortening lesser metatarsal osteotomy to aid the surgeon in determining the amount of bone needed to be resected to correct the deformity. This study was divided into three parts. In part I, 15 lesser metatarsals (metatarsals 2, 3, and 4) harvested from fresh frozen cadavers had shortening proximal osteotomies performed. This osteotomy removes a cylindrical piece of bone that is perpendicular to the metatarsal shaft from the proximal aspect of the lesser metatarsal to create axial shortening of the metatarsal and changes the relationship of the metatarsal head to the weightbearing surface. These metatarsals had five radio-opaque markers placed into them and were radiographed pre- and postosteotomy. These markers created a pre- and postgeometric graphic plotting for the changes in length, height, and dorsiflexion. Computer graphing was then utilized to analyze changes in height, length, and dorsiflexion of each metatarsal. Formulas were created from these plottings to determine the actual change in height, length, and dorsiflexion for a set amount of bone removed. The formulas created from these data were: Length: Actual change = Bone removed *0.95; Height: Actual change = Bone removed *0.54; and Dorsiflexion: Actual change = Bone removed *0.44 mm/deg. In part II of study, 15 identical saw bone lesser metatarsals were used to verify the formulas, by taking out the amount of bone needed for 0.5-mm increment change, starting at 1.0 mm and increasing to 8.0 mm. Techniques used were identical to part one. Part III was performed to demonstrate that the formula would be reproducible for height when there is a difference in the angulation of the metatarsal. Fifteen identical sawbones where plotted in plaster at declinations ranging from 8 degrees to 42 degrees. Then the osteotomy was performed removing 4.0 mm of sawbone from each specimen using the same technique as parts I and II. All parts and the formulas were statistically analyzed using a bivariate regression model, which showed that the formulas were valid for length, height, and dorsiflexion with a 95% confidence. With these experimental models, the authors found reproducible formulas that hopefully could aid the surgeon in determining the amount of bone they needed to resect to effect correction of this difficult reconstruction. PMID- 10789102 TI - Return to athletic activity after foot and ankle surgery: a preliminary report on select procedures. AB - One hundred thirty-eight "athletic" patients from the author's practice underwent retrospective review of their foot and ankle surgery that was performed from 1990 to 1997 to evaluate the time to return to activity. Athletes were defined as follows: professional, varsity college and high school, runners amassing more than 25 miles per week, or those involved in regular competition. No recreational athletes were included. Average follow-up for the group as a whole was 49.4 months, (range, 12-108 months). One hundred seventeen of the 138 patients were able to be contacted and/or had been evaluated by September 1998. One hundred thirteen patients considered their surgery a success. All but two patients were able to return to the desired level of performance. Twenty-three of the patients increased their activity level after surgery. Twenty-eight athletes underwent Achilles tendon-related surgery (average follow-up was 44.5 months). Runners undergoing peritenolyses had an average return to activity of 4.0 weeks (range, 3 6 weeks). Patients undergoing Achilles procedure involving bone resection had an average return to activity of 13.8 weeks (range, 10-20 weeks). Forty-four bunionectomy procedures were performed, including 31 first metatarsal osteotomies. The group's average follow-up was 52.9 months (range, 13-100 months), and average return to activity for the first metatarsal osteotomies was 8.9 weeks. There were 48 rearfoot procedures. Lateral ankle stabilization procedures returned to activity on an average of 10 weeks (range, 7-16 weeks), while excised ossicles (os tibiale externum, os trigonum) had an average return to activity of 9.1 weeks (range, 8-14 weeks). Seven neuroma patients (via dorsal approach) had a return to activity of 4.0 weeks (range, 2.5-6 weeks), sesamoidectomy 7.5 weeks (range, 4-10 weeks), and Valenti arthroplasty of 6.5 weeks (range, 4-12 weeks), respectively. There were minimal complications. Two patients in the Achilles surgery group required revision surgery. One patient with a bunionectomy had postoperative hallux limitus. Eight patients had symptomatic screws removed. PMID- 10789103 TI - Traumatic hallux varus repair utilizing a soft-tissue anchor: a case report. AB - Hallux varus is usually iatrogenic in nature; however, congenital and acquired etiologies have been described in the literature. The authors present a case of traumatic hallux varus secondary to rupture of the adductor tendon. Surgical correction was performed using a soft tissue anchor for maintenance of the soft tissues utilized for repair. PMID- 10789104 TI - Antibiotic beads in the treatment of diabetic pedal osteomyelitis. AB - Antibiotic-impregnated polymethylmethacrylate (PMMA) beads have improved the outcome of osteomyelitis treatment in both experimental models and clinical trials. The primary benefit of antibiotic-impregnated PMMA beads is that they provide high local concentrations of antibiotic while systemic levels of antibiotic remain low. Little has been written about the specific use of antibiotic-impregnated PMMA beads in the treatment of diabetic pedal osteomyelitis. The authors review antibiotic-impregnated PMMA beads and provide examples of their use in the treatment of diabetic pedal osteomyelitis. PMID- 10789105 TI - Foot deformity in frontometaphyseal dysplasia (Gorlin-Cohen syndrome): a case report. AB - A case of frontometaphyseal dysplasia (Gorlin-Cohen syndrome) in a child is described. The boy showed characteristic musculoskeletal changes, and had bilateral symptomatic cavovarus deformities of the feet. Surgical management of the foot deformities and the outcome are discussed. PMID- 10789107 TI - Stalking: ambiguous language can mask a crime. PMID- 10789106 TI - Evans calcaneal osteotomy. PMID- 10789108 TI - Re-visioning nurses' punitive attitudes within forensic psychiatric and correctional nursing: the significance of ethical sophistication. AB - 1. Development is an essential lifelong process if we are to compassionately care for patients in forensic and correctional nursing. 2. Many of us are blocked in our ability to understand and to care; we will need to grow to meet the challenges and to bear the shocks of forensic and correctional nursing. 3. Concepts that correspond to Kegan's levels of consciousness imply a progression from punitive attitudes toward more positive attitudes. PMID- 10789109 TI - HIV among people with mental illness: contributing factors, prevention needs, barriers, and strategies. AB - 1. Findings suggest that PWMI, including those with serious mental illness, are sexually active although some individuals, including mental health professionals, perceive them to be asexual or not sexually active. 2. PWMI may be particularly susceptible to HIV risk-related behaviors due to poor judgment, limited impulse control, deficits in problem-solving skills, and suicidal intent and self destructive behavior. 3. Nurses, due to their close interaction with PWMI, are in key positions to document HIV risk-related behaviors and to advocate for integrating HIV risk reduction strategies into the therapeutic milieu. PMID- 10789110 TI - Use of self-help manuals to treat problem drinkers. AB - In spite of research findings indicating that alcohol abusers may benefit from self-help manuals, health care providers have been slow to recognize their potential usefulness. As self-care strategies grow more popular and funding for traditional forms of alcohol treatment diminish, health care providers need to look more seriously at these materials. They also need to be aware of new innovations such as self-help materials that can be purchased on diskette. It may not be long before computerized materials sit next to printed manuals on bookstore shelves, and nurses need to know which materials to recommend. PMID- 10789111 TI - Post-traumatic stress disorder. A student nurse's personal experience of terror. AB - 1. Clients feel relieved when they have a label for what "ails" them. 2. Following vicitimization of one's self, exposure to a second traumatic event can trigger a reexperiencing of the original trauma. 3. Resolution of the trauma requires that the victim be able to formulate an explanation (supply meaning) as to why it occurred. PMID- 10789112 TI - 1998-based national population projections for the United Kingdom and constituent countries. AB - The 1998-based national population projections, carried out by the Government Actuary in consultation with the Registrars General, show the population of the United Kingdom rising from 59.2 million in 1998 to over 63.5 million by 2021. Longer-term projections suggest the population will peak around 2036 and then gradually start to fall. The population will become gradually older with the median age expected to rise from 36.9 years in 1998 to nearly 42 years by 2021. In 1998, there were 1.4 million (13 per cent) more children aged under 16, than people of pensionable age. However, by 2008, the population of pensionable age is projected to exceed the number of children. PMID- 10789113 TI - Population of households in England to 2021. AB - This article outlines the methodology and results of the Department of the Environment, Transport and the Regions (DETR) 1996-based household projections for England, and its regions. It examines differences between this round of projections and the previous 1992-based round. The 1996-based projections indicate that between 1996 and 2021 a growth of 3.8 million households can be expected in England, if recent trends continue. Both the national and subnational methodology and results are covered. PMID- 10789114 TI - Reviewing the mid-year population estimates in Northern Ireland. AB - This article describes the thorough review of the mid-year population estimates for Northern Ireland. No major problems have been identified with the current estimates but a number of enhancements have been put in place. The new figures are consistent with a number of administrative data sources. The overall effect has been to revise the estimated Northern Ireland population by about 6,000 people, or between 0.3 and 0.4 per cent, for each year between 1991 and 1997. The revised estimates for Northern Ireland have been incorporated into the population figures for the United Kingdom included in the Table section of this issue of Population Trends. PMID- 10789115 TI - Looking back to look forward: lessons from four birth cohorts for ageing in the 21st century. AB - Government projections indicate that by 2026 the number persons aged 60 and over living in Britain will reach 17.1 million, giving rise to concerns about the social and economic costs of an ageing population. Little attention, however, has been paid to the likely socio-economic characteristics of those retiring in the twenty-first century, and how they may differ from previous cohorts of elderly persons. This research analyses data from the General Household Survey (1974 1996) to examine the social and economic experiences of four birth cohorts. Findings indicate significant differences in the living arrangements, health and access to resources amongst the cohorts, which will have implications for the retirement prospects of the post-war baby boom generations in 2020 and beyond. PMID- 10789116 TI - Families, groups and clusters of local and health authorities: revised for authorities in 1999. AB - This is the second of two articles describing the revision of The ONS classification of local and health authorities of Great Britain to take account of area changes in the 1990's. It presents local authorities in Great Britain as at April 1999 in terms of Families, Groups and Clusters, based on 37 socio economic and demographic variables from the 1991 Census. Health authorities are presented in terms of Families and Groups only, based on the same variables. The previous article (in Population Trends 98) presented, for each local and health authority, up to four other authorities to which it is 'most similar'. PMID- 10789117 TI - [Virtual reality in radiology. Between hope and dilemma]. PMID- 10789118 TI - [Virtual reality in neurosurgery]. AB - DEFINITION: Virtual reality enables users to immerse themselves in a virtual three-dimensional world and to interact in this world. The simulation is different from the kind in computer games, in which the viewer is active but acts in a nonrealistic world, or on the TV screen, where we are passively driven in an active world. In virtual reality elements look realistic, they change their characteristics and have almost real-world unpredictability. USE OF VIRTUAL REALITY: Virtual reality is not only implemented in gambling dens and the entertainment industry but also in manufacturing processes (cars, furniture etc.), military applications and medicine. Especially the last two areas are strongly correlated, because telemedicine or telesurgery was originated for military reasons to operate on war victims from a secure distance or to perform surgery on astronauts in an orbiting space station. In medicine and especially neurosurgery virtual-reality methods are used for education, surgical planning and simulation on a virtual patient. PMID- 10789119 TI - [Computer-assisted oral, maxillary and facial surgery]. AB - BACKGROUND: Methods from the area of virtual reality are used in oral and maxillofacial surgery for the planning and three-dimensional individual simulation of surgeries. SIMULATION: In order to simulate complex surgeries with the aid of a computer, the diagnostic image data and especially various imaging modalities (CT, MRT, US) must be arranged in relation to each other, thus enabling rapid switching between the various modalities as well as the viewing of mixed images. Segmenting techniques for the reconstruction of three-dimensional representations of soft-tissue and osseous areas are required. We must develop ergonomic and intuitively useable interaction methods for the surgeon, thus allowing for precise and fast entry of the planned surgical intervention in the planning and simulation phase. SURGERY: During the surgical phase, instrument navigation tools offer the surgeon interactive support through operation guidance and control of potential dangers. This feature is already available today. Future intraoperative assistance will take the form of such passive tools for the support of intraoperative orientation as well as so-called tracking systems (semi active systems) which accompany and support the surgeons' work. The final form are robots which execute specific steps completely autonomously. DISCUSSION: The techniques of virtual reality keep gaining in importance for medical applications. Many applications are still being developed or are still in the form of a prototype. However, it is already clear that developments in this area will have a considerable effect on the surgeon's routine work. PMID- 10789120 TI - [Intraoperative navigation in paranasal sinus surgery with the Philips "Neuroguide" system]. AB - OBJECTIVE: To investigate the feasibility, precision and usefulness of computer aided surgery in ENT-sinus surgery. MATERIAL AND METHODS: 5 Patients with chronic sinus pathology and an indication for sinus surgery were elected. For intraoperative navigation we used a Philips "NEUROGUIDE" system and surgical instruments with LED's. Navigation procedures are described in detail in the paper, the system's precision was measured by pointing at anatomical landmarks. The accuracy was measured as the distance in millimeter between the bony structures of the CT scan on screen and the haircross of the pointer's tip on the screen. Another parameter of the systems accuracy was calculated by the system itself as the root mean square error in millimeter (RMSE) between the registered markers position and the marker position in the CT data set. RESULTS: Axial 3/3/1 mm spiral CT provided sufficient resolution, data transfer via optical disk was practicable. Positioning of the navigation equipment required some experience and the registration of the patients head position needed attention, as the markers have to be pointed at precisely. During operation, the head tracking system must not change its position on the patients head to ensure a correct navigation display. The main advantage of the computed navigation system was the constant orientation during the sinus surgical procedure. Frontiers and critical anatomical structures could be identified in the corresponding CT data set, thus enabling the surgeon to decide on the further procedure. At present stage, the operation time was increased through the handling of the navigation system for at about 15 min, resulting in additional time of narcosis. CONCLUSION: We found the computed navigation system Philips "NEUROGUIDE" system to be an established technical aid, ready to use for ENT sinus surgery, in our cases with a precision between 1 and 3 mm. These results were similar to results obtained with a SPOCS Navigation System from Aesculap, as previously published by us [17]. PMID- 10789121 TI - [Virtual endoscopy of the upper airway with spiral CT]. AB - PURPOSE: The technical conditions, the optimized scanning protocols and the facilities of virtual endoscopy (VE) are presented in an overview. METHODS: Phantom studies showed that, for a single-row-detector helical CT, collimation of 3.0 mm combined with pitch of 1.5 provided an optimal compromise between image quality and the scan duration per breath-hold. A multi-row-detector helical CT requires only a fraction of the scanning time. This is especially important for patients with dyspnea. RESULTS: The threshold-dependent virtual endoscopic surface rendering is a reliable and rapidly practicable reconstruction algorithm for the imaging of the upper airway. CONCLUSIONS: The VE technique is suitable for the imaging of space-occupying tumors and restricted stenoses. With VE follow up examinations can be performed non-invasively and interventional procedures can be prepared in an optimal way. Because of the computed tomographic data acquisition the structure of the mucosa is not assessable and there is no opportunity to obtain a biopsy specimen. PMID- 10789122 TI - [Construction and use of a virtual anthropomorphic phantom of the bronchial tree]. AB - MOTIVATION: Usually, phantoms are used to quantify properties of computer algorithms working on spiral CT data. The high complexity of the human bronchial tree, however, complicates the construction of a physical phantom enormously; hence, a virtual phantom must be employed. METHODS: Because the human bronchial tree is fractal, Lindenmayer systems were utilized to generate its structure. Applying the correct diameter and length ratios and branching angles, a virtual anthropomorphic phantom could be generated by a three-dimensional turtle graphic. The simulation of a spiral CT measurement on the virtual phantom delivered synthetic spiral CT data. RESULTS: The computer-controlled growing process supplies exact morphologic and morphometric data. The spiral CT simulation saves the time-consuming and complicated generation of a physical phantom. PERSPECTIVE: The virtual phantom can be completed by the introduction of blood vessels and lung parenchyma partitioned into lobes and segments. The field of application can thus be extended. PMID- 10789123 TI - [Use of virtual reality for MRI data of complex vascular structures]. AB - During the last years because of the progress in magnetic resonance imaging (MRI) magnetic resonance angiography (MRA) has become a serious alternative to conventional X-ray angiography. The potential of MRA in combination with methods for three-dimensional reconstruction will be presented and different methods for image post-processing are discussed based on a number of cases. The examinations were performed on a clinical 1.5 T magnetic resonance tomograph (Siemens Vision, Erlangen) using conventional MR angiography sequences. The different options of post-processing were carried out online on the console and offline using dedicated workstations (Siemens Virtuoso and CHILI). DISCUSSION: Complex post processing procedures are applied to different areas like pulmonary vasculature, thoracic aorta, abdominal aorta, and renal transplant arteries. Different diagnostic values can be seen for the variety of three-dimensional reconstruction methods. According to our experience volume rendering has been selected as the method of choice due to the time needed for reconstruction and the information content of the resulting image. PMID- 10789124 TI - [3D models for diagnosis and treatment planning in cardiology]. AB - Due to the development new of imaging devices which produce a large number of tomographic slices, advanced techniques for the evaluation of the large amount of data are required. Computer supported extraction of dynamic 3D-models of the patients anatomy from temporal series thus is highly desirable. Since the diagnostician should be able to quickly make sensible decisions based on the models, high accuracy is required within a minimum of time. We present modeling and visualization techniques that are realized within the Cardiac Station. Results for the application of these techniques to cardiac image data demonstrate their usability. Besides giving information about the patients morphology functional parameters can be derived from the data and visualized together with the model. In order to verify the model with the original image data and for the planning of real intervention interaction techniques are presented. PMID- 10789125 TI - [Dynamic MR mammography. Multidimensional visualization of contrast medium enhancement in virtual reality]. AB - BACKGROUND: The purpose of this study was the development of a method for fast and efficient analysis of dynamic MR images of the female breast. The image data sets were acquired with a saturation-recovery turbo-FLASH sequence which enables the detection of the kinetics of the contrast agent concentration in the whole breast with a high temporal and spatial resolution. In addition, a morphologic 3D FLASH data set was acquired. METHODS: The dynamic image datasets were analyzed by a pharmacokinetic model which enables the representation of the relevant functional tissue information by two parameters. In order to display simultaneously morphologic and functional tissue information, we developed a multidimensional visualization system, which enables a practical and intuitive human-computer interface in virtual reality. DISCUSSIONS: The developed system allows the fast and efficient analysis of dynamic MR data sets. An important clinical application is the localization and definition of multiple lesions of the female breast. PMID- 10789126 TI - [Individual planning of liver surgery interventions with a virtual model of the liver and its associated structures]. AB - BACKGROUND: In an interdisciplinary approach, HepaVision (MEVIS, Bremen), a software tool specifically developed for 3D visualization of the liver, was employed for individual planning of extensive liver resections and evaluation of living-relative donations. So far there is experience with more than 50 biphasic spiral CT examinations. RESULTS: The spatial relationship of large tumors to crucial hepatic structures, the demonstration and evaluation of anatomic variants regarding vascular supply and the risk stratification of liver failure by volumetric analysis on the basis of portal venous drainage supported precise indication for surgery. Surgical safety is increased by preoperative planning and simulation of necessary vessel reconstructions. By hiding selective areas of portal venous drainage or applying freely selectable clip planes, segmental as well as non-anatomical resections can be simulated and their effects analyzed. The virtual preoperative situs was confirmed intraoperatively in all 17 patients of our study population who underwent segmental liver resection for either a tumor or living-relative donation. PMID- 10789127 TI - [Virtual coloscopy with multi-slice computerized tomography. Preliminary results]. AB - SUBJECT: Using multi-slice computed tomography (MSCT) large body areas can scanned with high spatial resolution. In this study, MSCT was employed for virtual colonoscopy in various pathologies of the colon. MATERIALS AND METHODS: For MSCT a Somatom Plus 4, Volume Zoom scanner, (Siemens, Forchheim) was employed, equipped with four parallel detector rows. Twenty-five patients were included in this trial, prior to colonoscopy or partial colon resection. After cleaning the colon, distension was achieved by insufflation of room air using a rectal tube. The parameters of acquisition and reconstruction were as follows: collimation 4 x 1 mm and 4 x 2.5 mm respectively; tube charge per slice: 140-160 mAs; pitch 5-6; i.v. contrast medium: 120 ml Ultravist 300 (Schering) with a flow rate of 3 ml/s; delay: 35 s. For 3D reconstruction we used edge-enhanced volume rendering, virtual colonoscopy and extraluminal views of volume-rendered images. RESULTS: Nine polyps and four of five colon carcinomas were detected using MSCT virtual colonoscopy. In three patients with ulcerative colitis virtual coloscopy revealed morphological alterations compatible with this disease. In two of four patients with multiple diverticula of the colon the true extent of the disorder could be assessed in virtual colonoscopy. CONCLUSION: Utilizing virtual MSCT colonoscopy polyps and cancer of the colon can be reliably detected, if proper cleaning and distension is provided. On axial images alone smaller polyps may be assessed. The high z-axis resolution of MSCT offers superior conditions for CT based virtual colonoscopy. PMID- 10789129 TI - [VRATS--Virtual Reality Arthroscopy Training Simulator]. AB - The subject of this paper is a highly interactive medical training system for arthroscopic surgery; this is based on computer graphics and virtual reality (VR) techniques and offers an alternative to conventional training methods. To provide the virtual environment, a realistic 3D representation of the knee joint is derived from 2D medical image data. The use of tracking techniques guarantees an intuitive handling of the surgical instruments. The system allows navigation via a virtual camera and interaction with the virtual anatomical structures. First approaches for the simulation of tissue deformation caused by collisions with the instruments are implemented. One important advantage over conventional training systems is the possibility of verifying the training progress. Work is in progress on the realization of tactile feedback with the aim of providing a higher degree of interactive realism. PMID- 10789128 TI - [Virtual reality in MR colonography]. AB - Early detection and subsequent removal of colorectal polyps have been shown to constitute an effective approach for decreasing the incidence of colorectal cancer. The lack of an ideal modality for colorectal polyp screening stimulated interest in the development of CT-colonography and MR-colonography. Both techniques allow the colon to be analyzed in a cross-sectional as well as a virtual endoscopic format. Causing no side-effects and not concerning for radiation exposure MR-colonography warrants further consideration. Additional to detecting polyps down to 6 mm in size the inner wall contour and the morphology of the colonic wall itself can be assessed. New developments like fecal tagging will increase patients acceptance comparing to other diagnostic techniques. In search of an ideal modality for polyp screening MR-colonography will become a potent option in the diagnostic arsenal. PMID- 10789130 TI - [Free hand acquisition, reconstruction and visualization of 3D and 4D ultrasound]. AB - 3D Ultrasound will find in the next years a wide popularity under the medical imaging applications. The method expands the well-known sonography on the third dimension, therefore it becomes possible to generate spatial 3D views of internal organs. It is further possible to display static (3D) as well as dynamic organs (4D, e.g. pulsating heart). The clarity of the three-dimensional presentation supports very effectively the navigation. In this article we review the upgrading of conventional ultrasound devices on 3D and 4D capabilities, as well as the display of the datasets by corresponding visualisation and filtering approaches. PMID- 10789131 TI - [Basic principles of data acquisition and data processing for construction of high quality virtual models]. AB - Creating models for virtual reality subdivides into several steps. The aim of the data acquisition is the extraction of nearly isotropic (same solution in all three axes) data sets with low noise content. An approximate isotropy can be achieved by suitable choice of scan parameters. For raw data reconstruction, the application of high-resolution reconstruction algorithms is prohibited due to increased noise. A missing isotropy can computationally be approximated by interpolation. Further noise suppression is achieved by applying filters. Additionally, the contrast of the object for segmentation can be increased by image processing operators. The correct choice of the segmentation method and the editing tools is essential for a precise segmentation with minimal user interaction. Prior to visualization, smoothing the shape of the segmented model (shape-based or morphological interpolation, polygon reduction of wire frame model) further improves the visual appearance of the 3D model. PMID- 10789132 TI - [Evaluation of diagnostic scores via intercategorial kappa coefficients]. AB - A study design for the evaluation and confirmed modification of diagnostic rules is described. Based on the assessment of intercategorial rater agreement, those categories, which cause confusion in multiple ratings, can be combined and the gain in rater agreement can be assessed by the computation of appropriately modified kappa measures. The proposal is illustrated by the analysis of an evaluation trial related to SPECT based classification of patients with Graves' ophthalmopathy. PMID- 10789133 TI - [Undefined retroperitoneal and intrahepatic space-occupying lesions]. PMID- 10789134 TI - [Chronic inflammatory bowel diseases: Crohn disease and ulcerative colitis. 1: Etiology and pathogenesis, diagnosis. Diagnostic imaging in Crohn disease]. PMID- 10789135 TI - [Reduced neuronal nitric oxide synthetase and c-protein kinase levels in Alzheimer's disease]. AB - INTRODUCTION: Alzheimer's disease is characterized by a general and progressive dementia and by the presence of beta-amiloide deposits. OBJECTIVE: The levels of neuronal nitric oxide synthase (NOS) and protein kinase C (PKC), and the relationship between these proteins, the free-radical theory and the high level of beta-amiloide in Alzheimer's disease, have been studied. MATERIAL AND METHODS: The study has been performed in samples of Alzheimer's disease (superior, medial and inferior regions of temporalis gyrus) from control individuals and patients. The tissue was homogenized and the proteins were analyzed using monoclonal antibodies for NOS and PKC. RESULTS: Lower levels of neuronal constitutive NOS (37% +/- 2.5 and 52% +/- 3.0) in Alzheimer's disease derived superior and inferior temporalis gyrus, respectively, were observed. No changes were found in superior temporalis gyrus in PKC isoforms levels, involved in the processing for the beta-amiloide precursor protein. In the medial and inferior regions the PKC level was 5% +/- 0.5 to 22% +/- 3.0. CONCLUSIONS: These results could be related with an imbalance in the superoxide/nitric oxide ratio as a consequence of the non-inhibition of lipoxygenase, and with a neurotransmission dysfunction due, all this, to the decrease of nitric oxide levels. On the other hand, a relationship could be proposed between the high concentrations of beta-amiloide and the decrease in PKC levels in Alzheimer's disease. PMID- 10789136 TI - [Disseminated acute encephalomyelitis in children. A retrospective study of seven patients]. AB - INTRODUCTION AND OBJECTIVE: Acute disseminated encephalomyelitis (ADEM) is an inflammatory demyelinating disease of the central nervous system. This study is based on clinical symptoms and diagnostic tests employed. PATIENTS AND METHODS: We describe a seven children series indicating the initial neurologic abnormalities, diagnostic tests, treatments used and clinical-neuroradiological evolution. RESULTS: The mean presentation age was 4.1 years. Initial neurologic symptoms were mainly spastic hemi/paraparesis, cerebellous and pyramidal syndrome, consciousness changes, meningeal signs, seizures and cranial nerve palsies. The cerebrospinal fluid was abnormal in four patients with positive serologic tests in two of them (Coxsackie B). Electrophysiological studies were affected in 50%. MRI findings consisted of multifocal supratentorial white matter lesions. Clinical evolution revealed a progressive improvement with resolution after two months. Follow-up was made between six months and five years. The treatment was based on aciclovir and corticosteroids. CONCLUSIONS: ADEM runs a monophasic course of progressive neurologic abnormalities. Diagnosis is based on suggestive clinical and neuroimaging findings. Generally speaking, MRI showed resolution of multifocal lesions in conjunction with clinical improvement. PMID- 10789137 TI - [Conventional electroencephalography and delirium]. AB - OBJECTIVE: To review the electroencephalographic changes in conventional tracings made in patients with an acute attack of delirium described in the literature. PATIENTS AND METHODS: These alterations are considered in a sub-sample of 60 patients randomly selected from a sample of 223 persons admitted to hospital for different medical and surgical conditions and seen in psychiatric consultation for delirium during the years 1996 and 1997 in the Hospital General Yague in Burgos, Spain. The methods used included a conventional EEG tracing, following the usual clinical technique, in the context of a prospective, observational study. The changes were classified according to the trace seen and overall criterion of the neurophysiologist. The results were compared with simple statistical tests (frequency and percentage counts, chi squared and residual contingency tables, average comparisons and unidirectional Anova variance analysis and Bonferroni post hoc test). RESULTS: Marked alterations of tracings were seen in 83.3% of cases (78.7% 'slight', 'moderate' and 'excessive' slowing and 6.6% 'rapid' anomalies. Statistically, 'multi-factorial' and 'on dementia' types of delirium (characterized on the electroencephalogram by different degrees of slowing of the recording) could be clearly differentiated from delirium due to abstinence from alcohol which was mainly seen with 'fast' recordings. CONCLUSION: It was confirmed that there are two distinct types of delirium which are very different from the electroencephalographic point of view. PMID- 10789138 TI - [Clinical course of post-laminectomy cervical spondylitic myelopathy]. AB - INTRODUCTION AND OBJECTIVE: To study the postsurgical clinical course of spondylitic myelopathy. PATIENTS AND METHODS: We assessed 39 patients admitted to the Departments of Neurology and Neurosurgery of the Hospital Clinico Quirurgico Hermanos Ameijeiras, Cuba, with a confirmed diagnosis of spondylitic myelopathy who had decompressing laminectomies during the period between January 1996 and December 1997. RESULTS: Of the predicted variables, there was predominance of an age under 60 years (51.3%); male sex (71.8%); a history of cervical trauma and habitual dangerous physical activity in 38.5% and 41% respectively. The duration of preoperative symptoms was less than one year in 46.2%. The vertebral level most often involved was C5-C6 (97.4%). In 51.3% three levels were affected: 100% of the patients had stenosis in the zone of maximum compression; 28.2% had congenital stenosis of the canal and 66.7% had partial block shown on myelography or magnetic resonance studies of the cervical spine. There was clinical improvement in 78.4% after one month, in 91.9% after 3 months and in 94.6% after 6 months; only 2 patients had not improved on completion of the study. Motor and sensory function of the legs improved more than motor function of the arms and sphincter. CONCLUSIONS: The variables which influenced the degree of improvement were: age, duration of the symptoms, number of vertebral levels affected, presence of blockage and functional state. After application of the multiple regression model, it was seen that the duration of preoperative symptoms is an important predictor of the postoperative clinical course. PMID- 10789139 TI - [Dorsal medullary compression due to a primary vertebral chondrosarcoma]. AB - INTRODUCTION: Chondrosarcomas are the second commonest type of bony tumors, although they are not commonly found to be vertebral. They may originate in healthy bone or develop in a cartilaginous lesion with sarcomatous degeneration. CLINICAL CASE: A 62 year old woman complained of pain in the back and weakness of her legs. Following magnetic resonance studies a tumour at the level of the D6-D7 vertebrae was diagnosed. This was operated on and an extradural tumour, a grade 11 chondrosarcoma was removed. A local recurrence was removed two years later and three years afterwards she is still asymptomatic. CONCLUSIONS: A spinal chondrosarcoma is a tumour which usually presents with local pain and signs of spinal involvement. On magnetic resonance it is iso-intense on T1 and shows up with contrast. The treatment of choice is radical surgery since it does not respond well to oncological treatment. PMID- 10789140 TI - [Upward and downward gaze palsy with a unilateral mesencephalic hemorrhage]. AB - INTRODUCTION: Acute vascular lesions limited to the midbrain can yield varied disturbances of eye movements. They are more often infarcts than hemorrhages. CLINICAL CASE: We present the case of a 49 year old, non-hypertensive man who presented with sudden onset of bilateral up and down gaze palsy. CT defined a unilateral hemorrhage in the right mesencephalic tegmentum. CONCLUSION: Cases of upward and downward gaze palsy in association with unilateral upper midbrain hemorrhage are very uncommon. We present a new case. PMID- 10789142 TI - [Recent advances in genetics of epilepsy. Genetic of mitochondrial epilepsy]. AB - INTRODUCTION: Recently the molecular basis of a series of clinical disorders associated with defects in the oxidative phosphorylation system (OXPHOS system) leading to ATP synthesis, the final pathway of mitochondrial energy metabolism, has been established. The polypeptide components of the OXPHOS system are codified in both nuclear and mitochondrial DNA. Therefore these mitochondrial diseases may be originated by mutations of genes found in both genetic systems. DEVELOPMENT: In recent years, several such neuromuscular diseases have been defined and associated with mitochondrial DNA mutations. One of the most striking of these is the syndrome of myoclonic epilepsy with ragged red fibres (MERRF), characterized by myoclonic epilepsy of maternal inheritance. This disorder is caused by a specific mutation on the mitochondrial tRNA(Lys) (position 8344), which gives rise to a reduction in the level of lysil-tRNA(Lys) and thus to premature termination of the translation of proteins codified in the mitochondrial DNA. PMID- 10789143 TI - [Prognosis of epilepsy and suppression of treatment. Prognosis of epilepsy]. AB - OBJECTIVE: On the occasion of the XXXVII Annual Meeting of the Spanish Anti Epilepsy League we have reviewed the prognosis of epilepsy. DEVELOPMENT: The latest population studies of prognosis indicate that the long-term prognosis of epilepsy is good. Over 80% of these patients experience remission of their illness for at least three years, within nine years of diagnosis. The probability of remission is little influenced by the aetiology of the epilepsy. The mortality of epileptic patients is 2.5-3 times greater than that of the general population, adjusted for age, is markedly influenced by the aetiology and generally occurs within the first few years after diagnosis. PMID- 10789141 TI - [Subdural empyema due to Mycoplasma hominis following epidural anesthesia]. AB - INTRODUCTION: In the literature there are sporadic reports of spinal epidural abscesses after epidural anaesthesia (Staphylococcus aureus in 82%), whilst subdural empyemas are more often related to ear and sinus conditions. CLINICAL CASE: A 32 year old woman with a clinical history of migraine and symmetrical frontal atrophy on a previous cerebral CT scan, after Caesarean section under epidural anaesthesia, presented with orthostatic headache two days later. On the fourth day it had become constant and she had a high temperature which was considered to be caused by infection of the surgical wound. Neurological examination was found to be normal, the CT scan was inconclusive and the CSF showed a lymphocytic pleocytosis without consumption of glucose. In view of her worsening clinical condition on the ninth day, in the absence of a cutaneous focus and on suspicion of a para-meningeal infective focus, lumbar MR was done and found to be normal, and cerebral MR which showed images compatible with a right fronto-parietal subdural empyema. After a parietal craniotomy and culture of the surgical specimen, colonies of Mycoplasma hominis were grown, similar to those grown from the exudates of the abdomical surgical wound. Treatment was started with ciprofloxacine. CONCLUSION: We consider that following epidural anaesthesia the patient developed hypotension of the CSF with a secondary subdural hematoma or hygroma and this became infected by hematogenous spread of Mycoplasma hominis. PMID- 10789144 TI - [Pharmacological basis for withdrawal of antiepileptic drugs]. AB - OBJECTIVE: To review the pharmacological basis for withdrawal of antiepileptic drugs: the mechanisms by which seizures reappear, aspects of treatment which affect relapses and procedures for withdrawal of medication. DEVELOPMENT: Antiepileptic drugs are not curative, so when they are withdrawn the natural course of the condition becomes evident. Reappearance of seizures may be due to lack of protection and/or an abstinence syndrome. Seizures due to lack of protection occur following withdrawal of any antiepileptic drug when the epilepsy is not cured; they may not reappear for years (although over 80% occur within a year) and treatment then has to be restarted. They seem to be less frequent after withdrawal of carbamazepine or phenytoin than after withdrawing valproate, although the reason for this is not understood. Seizures due to an abstinence syndrome only occur after withdrawing benzodiazepines, phenobarbitone and primidone; they are seen in patients with both active and inactive epilepsy whilst the drug is being withdrawn and tend to be self-limiting. It is not necessary to reintroduce the drug when epilepsy is cured. Felbamate and vigabatrin cause seizures related to their withdrawal, but the mechanism of this is not clear. There is no scientifically established guideline for withdrawing antiepileptic drugs, but it is considered important to stop one at a time, starting with those which may cause abstinence syndromes, followed by the more toxic, less effective antiepileptic drugs, which cause more drug interactions and are more awkward to take. CONCLUSION: Further specific studies are necessary to establish the mechanisms of relapses and the scientific basis for withdrawal of antiepileptic drugs. PMID- 10789145 TI - [Prognosis of epilepsy and withdrawal of treatment: withdrawal of treatment in childhood and adolescents]. AB - OBJECTIVE: To assess the pharmacodynamic principles of withdrawal of long term treatment in children and adolescents with epilepsy. DEVELOPMENT: After considering the clinical (toxicity of antiepileptic drugs, relapses of seizures when long term treatment is stopped), social and economic factors which justify withdrawal of long term treatment, the best time to start the withdrawal of treatment is decided, bearing in mind factors related to the patient, his illness and the antiepileptic drugs themselves. PMID- 10789146 TI - [Withdrawal of antiepileptic treatment in adults]. AB - INTRODUCTION: The withdrawal of antiepileptic treatment is a major challenge for the epileptologist who has to make a decision involving two risks, namely long term maintenance of drug treatment and recurrence of the seizures. OBJECTIVE: In our review we wish to systematize the process of decision-making and calculation of the possibilities of recurrence. DEVELOPMENT: First we review the factors to be considered when making a decision. This is complex since epileptics are a very heterogeneous group of patients. We emphasize the respect due to the patient who, in the end, is the one who assumes the risk involved. Then we consider the factors to be taken into account when calculating the individual's possibilities of having a recurrence and describe a theoretical outline of non-recurrence. Finally, we give details of the practical aspects of withdrawal of treatment. CONCLUSIONS: We emphasize that the current challenge in this field is to design a method of measuring risks to permit the prediction of probabilities of recurrence in individual patients. We quote examples of the classification and measurement of risks, and suggest that only by institutional measures--perhaps by organizations such as LECE--can further essential epidemiological studies, of great scientific interest in the long term, be carried out. PMID- 10789147 TI - [Neuroimaging in epilepsy. Advances in SPECT and PET in epilepsy]. AB - INTRODUCTION: The incorporation of neuroimaging techniques into the protocol for localization of epileptogenous foci has considerably reduced the need for use of intracranial electrodes. DEVELOPMENT: SPECT and PET are most useful in cases of epilepsy with no cerebral morphological anomalies, in which there are therefore no changes which shown on MR. Cerebral SPECT with perfused tracers, such as 99mTc HMPAO, 123I-MP or 99mTc-ECD, permit the study of patients during interictal and perictal phases. In the latter case, although the procedure is complex, one may observe an increase in cerebral blood flow in the epileptogenous focus in 90% of the patients. PET with FDG allows interictal study of patients, and shows reduced metabolism in the epileptogenic region. By comparison with video-EEG there are S = 84% and SP = 86% in temporal lobe epilepsy. The diagnostic efficiency of these two techniques is rather less in the localization of extratemporal foci and in multifocal epilepsies, although it is always better than with other diagnostic techniques. They have also been shown to be useful in the prognosis of functional recovery after surgery. CONCLUSION: The possibility of studying neuroreceptors may be of great use in the investigation of the etiopathogenesis of epilepsy, and thus lead to improvement in the clinical and therapeutic management of patients. With SPECT, 123I-Iomacenil and 123I-lododexetimide are used. With PET and 11C Carfentanyl opiate receptors have been studied. With 11C-Fluomacenil one may study the cerebral distribution of the benzodiazepine receptors. PMID- 10789148 TI - [New developments in clinical neurophysiology: magnetoencephalography. Contributions of magnetoencephalography to the study of epilepsy]. AB - INTRODUCTION: Magnetoencephalography (MEG) is a novel neurophysiological technique based on the detection of the tiny cerebral magnetic fields generated by the intracellular electrical currents. DEVELOPMENT: It offers several theoretical advantages to electroencephalography, like the transparency of the skull and soft tissues to the magnetic signal and the sensitivity to tangential fields generated in the cerebral sulci. Both properties allow an easier and more accurate modelling of the intracerebral sources. Furthermore, both techniques are complementary. On the other hand, its cost is the most limiting factor for its routine implementation in clinical practice. MEG is fully integrated with neuroimaging giving rise to the term magnetic source imaging. Integration with other neuroimaging modalities such us functional magnetic resonance imaging constitutes one of the major developments for the future. CONCLUSION: In this review, we present an overview of the physiological basis and technical aspects of magnetoencephalography, its comparison with electroencephalography, and its increasing number of clinical applications in the study of the different epilepsy syndromes as well as its role in epilepsy surgery. PMID- 10789149 TI - [Experimental models in epilepsy]. AB - INTRODUCTION: Epilepsy is an heterogeneous collection of neurological disorders that have in common a transient and recurrent hypersynchronous activation of large populations of neurons in distinct focal areas or in the entire brain. DEVELOPMENT: Research into the cellular mechanisms of epilepsy focuses on understanding. 1. The alterations on cellular and network excitability, and 2. The mechanisms of hypersynchronization. Several different animal models exist that mimic different types of focal and generalized epilepsy in humans. Hypothesis derived from such models is helping to design more effective therapies. PMID- 10789150 TI - [Hernia of the L1-L2 intervertebral disc in an elderly patient]. PMID- 10789151 TI - [Gerstmann syndrome caused by right parietal ischemic stroke]. PMID- 10789152 TI - [Heterotopic radiological calcifications in two brothers with pseudohypoparathyroidism]. PMID- 10789165 TI - New dental materials. PMID- 10789153 TI - [News about epilepsy surgery]. PMID- 10789166 TI - Mercury pollution in dental office waste water. PMID- 10789167 TI - Dental unit waterline contamination. PMID- 10789168 TI - Depends on where you sit! PMID- 10789169 TI - Accepting the need for "overkill" in infection control. PMID- 10789170 TI - Dental plans for the new millennium. PMID- 10789171 TI - Caries-detector dyes--how accurate and useful are they? AB - Commercially available caries-detector dyes are purported to aid the dentist in differentiation of infected dentin, yet research has established that these dyes are not specific for infected dentin. They are non-specific protein dyes that stain the organic matrix of less mineralized dentin, including normal circumpulpal dentin and sound dentin in the area of the amelo-dentinal junction. A considerable body of evidence indicates that conventional tactile and optical criteria provide satisfactory assessment of caries status during cavity preparation. There is reason for concern that subsequent use of a caries-detector dye would result in unnecessary removal of sound tooth structure. The use of caries-detector dyes has also been suggested as a diagnostic aid for occlusal caries. Although diagnosis of carious dentin beneath apparently sound enamel can be challenging, there is a lack of substantive evidence supporting the use of dyes for this purpose and false positives are a significant concern. Careful visual inspection combined with bitewing radiographic diagnosis has been shown to be the most reliable diagnostic method for the presence of infected dentin requiring operative treatment. PMID- 10789172 TI - Internal derangements of the temporomandibular joint: the role of arthroscopic surgery and arthrocentesis. AB - Arthroscopic surgery appears to be a safe, minimally invasive and effective method for treating internal derangements of the temporomandibular joint (TMJ), reducing pain and increasing mandibular range of motion for approximately 80% of patients. Although these results are encouraging, they are largely based on retrospective, uncontrolled and short-term studies. The landmark observation that lysis and lavage in only the upper compartment of the TMJ produce successful clinical results without repositioning the disc has prompted clinicians to question the importance of disc position as a significant factor in the etiology of TMJ pain dysfunction. Although there are prospective, controlled, randomized short-term studies indicating that arthrocentesis and arthroscopic surgery have comparable success rates in the management of acute TMJ closed lock, similar long term studies are lacking. Until they have been done, the roles of arthroscopic surgery and arthrocentesis in the management of TMJ internal derangements remain unclear. PMID- 10789173 TI - [Volume determination of pigment epithelium detachment in AMD by laser scanning tomography]. AB - BACKGROUND: The possibility of using 3D mapping of AMD-related RPE detachments by means of laser scanning tomography was evaluated to correlate the fluorescein and tomographic findings. METHODS: Sixty eyes with AMD-related RPE detachments of 55 consecutive patients (19 men, 36 women) between 54 and 87 years of age (mean: 72.2 years) were examined using the Heidelberg Retina Tomograph (HRT). The parameters considered were area, volume, maximal height and 3D configuration of the RPE detachments. The tomographic data were analyzed and correlated with the fluorescein angiographic findings. Follow-up examinations were done at 3 and 6 months later. RESULTS: The mean +/- SD area of elevation was 10.59 +/- 5.51 mm2 (range, 0.93-19.73), which correlated well with the angiographic measurements. The mean maximal height was 0.42 +/- 0.19 mm (range, 0.11-0.83), mean volume was 2.55 +/- 1.9 mm3 (range, 0.073-6.63). We found a tendency to grow for untreated RPE detachments, depending on the volume at the first measurement. Three RPE detachments of high volume (mean 0.501 +/- 1.3 mm3) resulted in tearing of the RPE. The angiographic findings of localized neovascularizations in the RPE detachment area (39 of 60 eyes) showed a corresponding irregularity of the surface in most of the correlating 3D HRT figures. CONCLUSIONS: Confocal laser scanning tomography allows analysis of 3D configurations and a quantitative measurement of RPE detachments in AMD. Therefore, this diagnostic technique appears to be useful, especially for differentiated follow-up examinations (as in therapy-control studies). Furthermore, the analysis of 3D configurations seems to be useful to estimate the risk of tearing of the RPE and may help to indicate underlying neovascularizations. PMID- 10789174 TI - [Erbium:YAG laser vitrectomy: temperature measurements in different replacement materials]. AB - BACKGROUND: The erbium:YAG laser has the potential of being used routinely for vitrectomy because of the excellent quality of liquefying vitreous structures and the low vacuum forces required. However, the use of silicone oil and perfluorocarbon may lead to unwanted temperature increases in the microsurgery probe. The aim of this work was to investigate this side effect. MATERIALS AND METHODS: Different replacement materials such as water, methocel, silicone oil and perfluorocarbon were used in a simple eye model. The temperature increase during laser application was measured by means of thermocouples. The maximum temperature increase and time decay were derived with and without aspiration from these time-resolved measurements. The average power at the distal end of the microsurgery probe was chosen to be 1 W. RESULTS: The temperature increase with aspiration in water was found to be significantly smaller than all other replacement materials. Interruption of aspiration leads to a critical temperature increase of approximately 14 K; however, this increase occurred very slowly (decay time 200 s). A comparable result could be observed for methocel, which was used to simulate condensed vitreous structures. In perfluorocarbon and silicone oil we measured a far higher increase in temperature of up to 130 K within a few seconds. Furthermore, small remnants of carbonized materials can be seen in the microsurgery probe after laser application. CONCLUSIONS: The temperature increase during erbium:YAG laser vitrectomy in water can be considered to be harmless for other intraocular structures. However, insufficient aspiration or increased vitreous condensation leads to increased temperature in the microsurgery probe. Our results demonstrate that the use of erbium:YAG laser vitrectomy in materials such as silicone oil or perfluorocarbons is contraindicated. PMID- 10789175 TI - [Errors in keratometry for intraocular lens implantation in infants]. AB - To evaluate investigator- and instrument-dependent errors of keratometry in children and their possible influence on the predicted postoperative refraction after intraocular lens implantation. METHODS: Keratometry readings of five children with congenital cataract were taken preoperatively under general anesthesia (age 1.5-7.5 months, mean = 4.4 months). Both eyes were measured by two investigators with a manual Zeiss keratometer and an Alcon portable automated keratometer. Three measurements were performed with each instrument. Axial length was determined with ultrasound and the SRK II-formula was used for intraocular lens calculation. RESULTS: The investigator-dependent variability was 0.22 mm +/- 0.07 SEM for the manual keratometer and 0.20 mm +/- 0.07 SEM for the automated keratometer. The instrument-dependent variability was 0.44 mm +/- 0.12 SEM for investigator A and 0.34 mm +/- 0.09 SEM for investigator B. These results suggest that deviation from the required postoperative refraction of up to 6.0 D has to be expected in individual cases if intraocular lenses are implanted. CONCLUSIONS: The lack of fixation in children who have keratometry under general anesthesia leads to inaccurate keratometry readings, which will cause rather high deviations of the precalculated postoperative refraction. Our results explain the high rate of residual refractive errors reported in the literature after intraocular lens implantation in children. In order to improve the accuracy of keratometry multiple measurements of corneal curvature should be taken. PMID- 10789176 TI - [Peribulbar anesthesia versus topical anesthesia in cataract surgery: comparison of the postoperative course]. AB - BACKGROUND: Phacoemulsification with topical anesthesia is a proven alternative to peribulbar or retrobulbar anesthesia. Application of lidocaine in the anterior chamber before surgery is one method to achieve good intraoperative analgesia. The purpose of this study was to find out whether there are any differences in the postoperative course between patients with peribulbar injection and patients with topical anesthesia. PATIENTS AND METHODS: A total of 186 patients scheduled to undergo elective cataract surgery were included in the study. The patients were randomly assigned to receive sponge-anesthesia and intraocular injection of 0.15 ml lidocaine 1% or a peribulbar injection of 6 ml Xylonest 2%. Uncorrected visual acuity was measured 30 min after surgery. On the first postoperative day, the cornea and ocular inflammation affecting the anterior chamber were examined with a slit lamp; best corrected visual acuity and postoperative pain were documented. RESULTS: Thirty minutes after surgery, the uncorrected visual acuity was significantly better in the lidocain group. At 1 day we saw no differences concerning visual acuity. Descemet folds occurred in 6.5% of the peribulbar group and 20.5% in the lidocain group. CONCLUSION: Topical anesthesia with sponge anesthesia and intraocular application of 0.15 ml lidocain 1% circumvents the complications of a peribulbar/retrobulbar injection. The patient profits from rapid visual rehabilitation. PMID- 10789177 TI - [CO2 laser in ophthalmology]. AB - BACKGROUND: Baker et al. used a CO2 laser for blepharoplasty for the first time. The ultra-pulse CO2 laser provides a sophisticated instrument that makes more application alternatives possible in ophthalmology. PATIENTS AND METHODS: From March 1997 to February 1998 surgical operations were performed in the eye clinics in Hagen and Darmstadt using the CO2 laser: 145 eyelid operations and 8 others (enucleation, evisceration, oral mucosa plastic surgery, secondary hydroxyapatite implantation, squint operation). In 34 patients in a total of 64 operations, after an average follow-up period of 7 (4-11) months, an examination was carried out. RESULTS: There were serious complications in only one case. Three weeks after transconjunctival fat resection and resurfacing, one patient developed an inflammatory reaction with resulting ectropion on both sides. During steroid therapy the left lower lid flattened, but on the right side a slight ectropion remained. CONCLUSION: The CO2 laser proved to be a very useful instrument for bloodless tissue-protective surgery without conspicuous scar formation. PMID- 10789178 TI - [Cogan I syndrome: clinical aspects, therapy and prognosis]. AB - BACKGROUND: Cogan I syndrome is a rare, inflammatory, systemic disease that is typically characterized by severe audiovestibular dysfunction and various inflammatory eye changes. PATIENTS AND METHODS: Between 1978 and 1996 ten patients with Cogan I syndrome were treated at the eye hospital, ENT clinic and medical clinic of the University of Tubingen. Retrospectively, the clinical picture, course of the disease and outcome were investigated by reviewing the charts of these patients. RESULTS: Patient ages ranged from 18 to 78 years (mean 37 years) at the time of disease manifestation. Eight patients were female. Inflammatory ocular changes were observed in nine patients in both eyes. Uveitis was found in six patients, keratitis in five, episcleritis in three and scleritis in two patients. Both patients with scleritis showed a severe, partially sight threatening course of the disease despite immunosuppressive therapy. Severe hearing loss was seen in all patients. Further manifestations of Cogan's syndrome included pericarditis associated with arthritis, and polyserositis in one patient, and fibromyalgia in two patients. CONCLUSION: In Cogan I syndrome, typically both eyes are affected by the disease process. Various ocular inflammatory changes may occur including uveitis, scleritis, keratitis and episcleritis. The visual prognosis is mostly good, although severe sight threatening scleritis may occur. PMID- 10789181 TI - [Med. in Germany. Interview with Professor Dr. Gronemeyers about his recent book "Med. in Germany: stand with future"]. PMID- 10789180 TI - [Current therapy aspects of intraocular tumors]. AB - The most frequent primary intraocular malignancies are uveal melanoma in adults and retinoblastoma in children. Genetic findings in uveal melanoma now allow a better risk-prediction with regard to metastatic disease. New treatment modalities like endo-resection, trans-scleral resection, proton beam irradiation and trans-pupillary thermotherapy are now being established in clinical routine. Management of retinoblastoma has changed during the last years considerably. In hereditary retinoblastoma external beam radiotherapy (EBR) results in a sixfold increased risk for the development of secondary, non ocular malignant tumors in these patients. New treatment regimens based on systemic chemotherapy were developed to replace EBR. In combination with chemotherapy there has been a continuing trend toward more conservative focal treatment for retinoblastoma. Indications and first results of these new treatment modalities are presented. PMID- 10789179 TI - [Studies on the stabilization of the cornea in rabbits]. AB - BACKGROUND: The mechanical stabilization of the cornea in keratoconus may delay progression of this disease. The cross-linking techniques optimized in corneas of enucleated porcine eyes were investigated under in vivo conditions in rabbits to estimate the biocompatibility and duration of the stiffening effect. METHODS: Twenty-eight rabbits were treated monocularly, the fellow eye serving as control. The epithelium was mechanically removed and 19 eyes were treated with riboflavin plus ultraviolet irradiation (365 nm, 2 mW/cm2) for 45 min and 9 eyes with 0.075% glutaraldehyde for 20 min. After treatment, the eyelids were sutured for 3 days. The healing process was controlled by slit-lamp examination and photographically documented. After 1 month, 20 animals and after 3 months 8 animals were sacrificed, the eyes enucleated, and the stress-strain relation of the corneas measured and compared to the fellow eye. RESULTS: The epithelium was closed after 4-5 days. The transparency of the corneas remained clear during follow-up, and there were no signs of inflammatory reaction. Stress for a strain of 6% was higher in the treated corneas by a factor of 1.3 +/- 0.66 (P = 0.319) in the glutaraldehyde group and by a factor of 1.6 +/- 0.75 (P = 0.0408) in the riboflavin group at 1 month, and by 1.3 +/- 0.48 (P = 0.07) at 3 months after treatment. CONCLUSIONS: The cross-linking technique using riboflavin plus UV irradiation is suitable for at least temporarily stiffening the cornea in vivo and seems to be a promising method for conservative treatment of keratectasia. PMID- 10789182 TI - [Posterior embryotoxon]. PMID- 10789183 TI - [Ocular involvement in graft versus host disease after bone marrow transplantation]. PMID- 10789184 TI - [Participation of multiresistant gram-negative rods in systemic infections of newborns treated in the Neonatal Clinics of the University Hospital in Krakow in 1993-1997]. AB - Results of blood cultures in 1187 newborns hospitalized in the Department of Neonatology, University Hospital in Krakow in furing the last five years were analysed. A trial of monitoring the presence of multi-resistant strains of Enterobacteriaceae with new mechanisms of resistance to antibiotics as well as analysed of their antibiotics susceptibility patterns were undertaken. Surprisingly high percentage of Gram-negative rods E. cloacae and K. pneumoniae strains resistant to the 3rd generation of cephalosporins and participating in septic complications in the investigated hospital environment was demonstrated. There were identified 53 strains of Enterobacter sp., 35 strains of Klebsiella sp. and 3 strains of E. coli extremely resistant to ceftazidine, accounting for 72%, 56% and 5% of all isolated of the given kind, respectively. Dynamic increase in a number of multiresistant strains of K. pneumoniae from 11.1% of isolates in 1993 to 83% in 1997 was shown as well as remaining on a very high level frequency of isolation of Enterobacter sp. rods capable of producing chromosomal cephalosporinases was demonstrated. Full efficacy of carbapenems and high usefulness of ciprofloxacine in the treatment of infections were confirmed. The authors emphasize necessity for constant monitoring of susceptibility to antibiotics in resistant strains as well as for liquidating their sources and ways of their transmission. PMID- 10789185 TI - [The efficacy of subterranean therapy in the salt chamber of Kinga Spa in Wieliczka for patients suffering from allergic rhinitis]. AB - 42 patients (27 women and 15 men aged 14-65 years) suffering from atopic and non atopic rhinitis were submitted for 24 days to the subterraneo-therapy in salt chambers of Kinga Spa in Wieliczka during the exacerbation of their illness. 20 patients suffered from pollen or dust mites allergy and the next 22 patients from non atopic rhinitis with recurrent nasal polyposis. In the first week and at the end of the therapy rhinoscopy, peak nasal inspiratory flow and smell were evaluated as well as daily symptoms score was noted in each treated patient. The results of performed observations indicate that unrelatively to the type of the rhinitis: 1. the nasal symptoms (running nose, sneezing, block of the nose) resolved directly during the stay of the patient in the salt chamber but the more constant effect was visible with the time of the therapy. 2. an increase in the peak nasal expiratory flow was connected with the decrease of nasal mucosal edema, 3. the smell disturbances disappeared in the patients together with the decrease of mucosal edema. The observed changes seem to be connected with the influence of the salt chambers' microclimate on the cells osmolarity disturbances in the nasal mucosa provoked by the non infectious chronic inflammation. PMID- 10789187 TI - [Analysis of basement membranes status in skin squamous cell carcinoma (SCC)]. AB - The aim of study was analysis of basal membranes in skin squamous cell carcinoma (SCC) basing on immunoreactivity of basal membrane component--laminin. The study was carried out on 43 samples of skin SCC which were evaluated histopathologically with regard to 3-grade scale (G) of malignancy. In each case immunohistochemical reactions by use of ABC method were carried out in order to detect laminin within basal membranes. The amount of laminin in basal membranes was evaluated semiquantitatively thus enabling to distinguish 3 groups of SCC: with abundant, moderate and small amount of laminin. The obtained results revealed significant differences within basal membranes according to malignancy grade. Most frequent anomalies referred to differences in basal membranes regularity, thickness and quantitative changes. In group of cancers with abundant laminin the number of tumor nests surrounded with mostly well formed basal membranes was found. Cancers with moderate laminin revealed variously formed basal membranes with distinct morphologic alterations and local gaps in continuity. In group of cancers with small laminin solely remnants of basal membranes were found. The results of present study indicated predominance of moderate amount of basal membranes in skin G1-2 SCC and small amount of basal membranes in poorly differentiated G3 SCC. The use of immunohistochemical methods in evaluation of laminin expression can be a useful tool in routine histopathologic examination of tumors of epithelial origin. PMID- 10789186 TI - [Interleukin 3 (IL-3) in diagnosis and monitoring of non-small-cell lung cancer]. AB - Several circulating tumour markers for non-small-cell lung cancer (NSCLC) have been identified. Recent studies have focused on a new family of markers- hematopoietic growth factors. Some clinical investigations have shown cell surface receptors for interleukin 3 (IL-3) in lung cancer cells and autologous production of IL-3 in various human cell lines derived from NSCLC. The purpose of this investigation was to compare serum levels of IL-3 in non-small-cell lung cancer with a control group, to assess pre- and post treatment levels of IL-3 in relation to levels of commonly accepted tumour markers such as carcino-embryonic antigen (CEA) and cytokeratin fragment 19 (CYFRA 21-1), and to define the diagnostic sensitivity and specificity of IL-3 in NSCLC. In this study, the serum level of tumour markers was measured in 34 patients with NSCLC and in 20 healthy subjects. Serum samples were drawn before surgery and 10, 30, 90, 180 and 270 days after surgery. IL-3 and CEA were assayed using ELISA system and CYFRA 21-1 was measured by radioimmunoassay (RIA). Preoperative level of IL-3 was significantly increased in cancer patients relative to the control group. Concentrations of tumour markers were decreased after surgery and then increased (IL-3 and CEA) during chemio- or radiotherapy. The diagnostic sensitivity of IL-3 was 44% and the diagnostic specificity--85%. This investigation is one of the first studies assessing serum levels of IL-3 in the cancer patients. These results suggest that IL-3 may be useful in diagnostics of NSCLC, but this subject needs further studies. PMID- 10789188 TI - [Usefulness of bioelectric impedance as a method for evaluating body composition of patients on peritoneal dialysis]. AB - The aim of a study was to evaluate the usefulness of bioelectric impedance as a method of body composition analysis in patients treated with CAPD, with the special attention paid on hydration status and lean body mass. The values of parameters obtained by bioelectric impedance and other methods were compared. The impact of peritoneal dialysis fluid in peritoneal cavity on bioimpedance measurement results were also analyzed. The study was performed in 33 patients dialyzed with CAPD for mean period of 12.3 months, aged between 23 and 72 years (mean 50.9 years). Bioimpedance measurements were also performed in 10 healthy volunteers. The significant impact of 2-liters dialysate volume on measurement results was found. The percentage water contain as well as LBM are under this condition higher (59.2% vs. 58.3%; p < 0.005 and 80.73 vs. 79.6; p < 0.01, respectively), and body fat--lower (19.07 vs. 20.39%; p < 0.005) as compared to empty peritoneal cavity. We also found, that the values of body water obtained from BEI measurements are higher as compared to those calculated from Watson formulas and lean body mass values obtained from BEI analysis are higher as compared to those derived from creatinine kinetics (39.4 vs. 36.96 I.; p < 0.05 and 53.7 vs. 51.1 kg; p < 0.01, respectively). Total and lean body mass did not differ from the values predicted in the treatment group, however the percentage contain of body water was significantly higher (58.34 vs. 51.39%; p < 0.0001). No significant differences were found between body composition of CAPD patients and control individuals. PMID- 10789189 TI - Is CAPD a renal replacement therapy method of choice for women? AB - The aim of this study was to evaluate if there were any differences in clinical outcome as well as in values of quantitative adequacy/nutritional parameters in ESRD patients treated with continuous ambulatory peritoneal dialysis, depending on their sex. Nutritional and adequacy parameters: NPCR, Kt/V, weekly creatinine clearance (wClCr), dialysis index (DI), serum albumin concentration, as well as clinical parameters such as hospitalization rate, admission rate, peritonitis rate, exit-site infection rate and co-morbidity score were evaluated in 31 CAPD patients (12 F and 19 M). Lower comorbidity score (0.583 vs. 1.58 points; p < 0.05) and higher Kt/V total and residual (2.25 vs. 1.57; p < 0.01 and 1.7 vs. 1.42; p < 0.01, respectively) were found in women as compared to men. The value of the quantitative nutritional parameter--NPCR--was also higher in women (0.842 vs. 0.73 g/kg b.w./24 hours; p < 0.05). Despite these differences, only a small difference was found in the clinical outcome and survival between men and women. The obtained data may suggest that women can achieve better treatment results with the CAPD method as compared to men. PMID- 10789190 TI - [Variability of heart auscultation in patients with mitral valve prolapse]. AB - The aim of the study was to analyze the auscultatory findings and its variability in 84 patients (pts) with mitral valve prolapse syndrome (MVPS) who were examined in the different body positions. All the pts underwent the echocardiographical examination and in 32 subjects (38%) prolapse of the anterior mitral leaflet was found, in 40 pts (48%) prolapse of the posterior mitral leaflet and in 12 (14%) pts prolapse of the both mitral leaflets. The auscultation of the heart was done in the supine position, in the lying position on the left side and in the upright position of the patient's body. Characteristic for the mitral valve prolapse (MVP) auscultatory findings (midsystolic murmur, late-systolic murmur and/or mid systolic "non ejection" click) were demonstrated by 43 pts (51%) in the supine position. During the auscultation in the lying position of the pts on the left side, the auscultatory findings were found in 57 pts (68%) while in the upright position in 64 pts with MVPS (76%). The study showed that the auscultatory findings in pts with MVPS were demonstrated more frequently in the lying position of the body on the left side or in the upright position compared with the supine position. Moreover we found that 20 pts with MVPS (24%) did not demonstrate the characteristic auscultatory findings of MVP. PMID- 10789191 TI - [Risk factors for in-hospital course and long-term outcome in myocardial infarction]. AB - The aim of the present study is to separate the most important in-hospital and long term outcome risk factors in patients with myocardial infarction. We analysed 251 women and 630 men hospitalised for acute myocardial infarction between 1992-96. We compared history data, in-hospital course and long term observation within 2-6 years in a group of patients who died versus group of patients who survived. The most important risk factors of in-hospital death were: cardiogenic shock--with mortality rate--6.2, pulmonary oedema--2.8, ventricular fibrillation--2.7, third degree A-V block--2.5, supraventricular arrhythmia (atrial flutter, atrial fibrillation--2.4, previous myocardial infarction--2.4, diabetes--2.0, disturbances of intraventricular conduction--1.8. The most important risk factors of long term outcome were: congestive heart failure--III, IV class of NYHA at discharge--mortality rate--3.0, ejection fraction < 40%--2.7, disturbances of intraventricular conduction--2.2, in-hospital cardiogenic shock and/or oedema pulmonum--2.0, prior myocardial infarction--1.9, diabetes--1.7, in hospital ventricular fibrillation--1.6, supraventricular arrhythmia--1.6. Better predictors of survival we can obtain using multivariate analysis. This analysis allows to separate groups of patient with good, mean and poor prognosis which finally simplify choice of efficient kind of therapy. PMID- 10789192 TI - [The role of cellular glucose transporters in pathogenesis of diabetic nephropathy]. AB - The transport of glucose across plasma membranes is of paramount importance for the maintenance of cellular homeostasis and metabolism. Over the past few years it has been established that this process is mediated via a family of specialized and tissue-specific glucose transporters. It has been demonstrated that these facilitative glucose transporters may regulate the cellular uptake of glucose and consequently affect glucose metabolism. It has been suggested that increased utilization of glucose in glomerular cells results in the increased expression and activity of aldose reductase, protein kinase C and TGF-beta, which have been implicated in excessive extracellular matrix accumulation in diabetic nephropathy. In this report we review the identified forms of the glucose transporter family focusing on the systems expressed by the kidney. We also summarize the currently available experimental data suggesting that glomerular glucose transport systems may play a role in the development of diabetic nephropathy. PMID- 10789193 TI - [Endothelins--their physiological role and their clinical significance]. AB - Biochemical part of the paper presents the structure, biosynthesis and degradation of the endothelins. Physiological chapter deals with endothelin-1 endocrine and mitogenic action as well as its impact on diuresis, natriuresis and cardiovascular system. In the clinical part of the article the role of endothelin 1 in essential hypertension, congestive heart failure and chronic renal failure is described. PMID- 10789194 TI - [Hyperprolactinemia. II. Diagnosis and treatment]. AB - Implementing various diagnostic methods in the past some decades has allowed to improve knowledge on hyperprolactinemia. In the diagnostics of hyperprolactinemia, besides measuring blood levels of PRL and other hormones, there may be also helpful: stimulative tests with metoclopramide and TRH, CT scans, MRI, ophthalmological examination, etc. There tumors such as prolactinoma or other reasons leading to hyperprolactinemia. Establishing cause of hyperprolactinemia is important because of various treatment modalities available. PMID- 10789195 TI - [Icodextrin in peritoneal dialysis therapy]. AB - In the presented review paper we have shown a role of glucose polymers, including icodextrin, in the treatment of uraemia with continuous ambulatory peritoneal dialysis or cyclic continuous peritoneal dialysis. Glucose polymers as a component of peritoneal dialysis solution exert significant ultrafiltration during dialysis solution exchanges lasting 10-16 hpurs. This is especially advantageous in patients with high peritoneal permeability and results in prolongation of peritoneal dialysis treatment by several months. Dialysis solution containing glucose polymers sustains ultrafiltration during peritonitis. In this paper there are also described icodextrin kinetics after intraperitoneal administration, its influence on peritoneal transport and adverse effects observed in some patients using icodextrin solution. PMID- 10789196 TI - Information-accumulation theory of speeded categorization. AB - A process model of perceptual categorization is presented, in which it is assumed that the earliest stages of categorization involve gradual accumulation of information about object features. The model provides a joint account of categorization choice proportions and response times by assuming that the probability that the information-accumulation process stops at a given time after stimulus presentation is a function of the stimulus information that has been acquired. The model provides an accurate account of categorization response times for integral-dimension stimuli and for separable-dimension stimuli, and it also explains effects of response deadlines and exemplar frequency. PMID- 10789197 TI - The construction of autobiographical memories in the self-memory system. AB - The authors describe a model of autobiographical memory in which memories are transitory mental constructions within a self-memory system (SMS). The SMS contains an autobiographical knowledge base and current goals of the working self. Within the SMS, control processes modulate access to the knowledge base by successively shaping cues used to activate autobiographical memory knowledge structures and, in this way, form specific memories. The relation of the knowledge base to active goals is reciprocal, and the knowledge base "grounds" the goals of the working self. It is shown how this model can be used to draw together a wide range of diverse data from cognitive, social, developmental, personality, clinical, and neuropsychological autobiographical memory research. PMID- 10789198 TI - Time, rate, and conditioning. AB - The authors draw together and develop previous timing models for a broad range of conditioning phenomena to reveal their common conceptual foundations: First, conditioning depends on the learning of the temporal intervals between events and the reciprocals of these intervals, the rates of event occurrence. Second, remembered intervals and rates translate into observed behavior through decision processes whose structure is adapted to noise in the decision variables. The noise and the uncertainties consequent on it have both subjective and objective origins. A third feature of these models is their timescale invariance, which the authors argue is a very important property evident in the available experimental data. This conceptual framework is similar to the psychophysical conceptual framework in which contemporary models of sensory processing are rooted. The authors contrast it with the associative conceptual framework. PMID- 10789199 TI - The role of expectations in comparisons. AB - Expectations are proposed as a unifying framework for explaining a variety of findings on comparative judgment. Expectations may pertain to either the features of each separate object or their simultaneous occurrence in all relevant objects. Two experiments demonstrate that inducing expectations by varying the frequency with which a component occurs, as well as the frequency with which it is shared by the compared objects, results in augmented weighting of unexpected components. Expectations can arise not only from experienced frequencies but also from an inferred set of alternatives. Features of cohesive stimuli are shown to be more expected than features of noncohesive stimuli, in the latter sense. The relatively higher weighting of distinctive features in cohesive stimuli, as well as the higher weight assigned to structurally aligned differences, could thus reflect modifications in feature expectedness. Further experiments show how task dependent weighting interacts with expectations to produce asymmetries between similarity and difference judgments. Finally, it is argued that changes in expectations regarding objects' attributes can explain context effects across different domains, including some types of preference reversals. PMID- 10789200 TI - How persuasive is a good fit? A comment on theory testing. AB - Quantitative theories with free parameters often gain credence when they closely fit data. This is a mistake. A good fit reveals nothing about the flexibility of the theory (how much it cannot fit), the variability of the data (how firmly the data rule out what the theory cannot fit), or the likelihood of other outcomes (perhaps the theory could have fit any plausible result), and a reader needs all 3 pieces of information to decide how much the fit should increase belief in the theory. The use of good fits as evidence is not supported by philosophers of science nor by the history of psychology; there seem to be no examples of a theory supported mainly by good fits that has led to demonstrable progress. A better way to test a theory with free parameters is to determine how the theory constrains possible outcomes (i.e., what it predicts), assess how firmly actual outcomes agree with those constraints, and determine if plausible alternative outcomes would have been inconsistent with the theory, allowing for the variability of the data. PMID- 10789201 TI - The case against a criterion-shift account of false memory. AB - M. B. Miller and G. L. Wolford (1999) argued that the high false-alarm rate associated with critical lures in the Roediger-McDermott (H. L. Roediger & K. B. McDermott, 1995) paradigm results from a criterion shift and therefore does not reflect false memory. This conclusion, which is based on new data reported by Miller and Wolford, overlooks the fact that Roediger and McDermott's false-memory account is as compatible with the new findings as the criterion-shift account is. Furthermore, a consideration of prior work concerned with investigating the conditions under which participants are and are not inclined to adjust the decision criterion suggests that the criterion-shift account of false memory is unlikely to be correct. PMID- 10789202 TI - False memories and statistical decision theory: comment on Miller and Wolford (1999) and Roediger and McDermott (1999) AB - In an analysis of H. L. Roediger and K. B. McDermott's (1995) false-memory paradigm, M. B. Miller and G. L. Wolford (1999) argued that falsely recognized items occur because a bias toward calling such items "old" is created by their membership in a studied category. This interpretation was contested by Roediger and McDermott (1999). The authors of this article approach this issue as a statistical decision problem and observe that an explanation of false memory based on stored strengths and one based on decision process can have identical implications for data. Problems with equivalent formal models of this type can frequently be resolved by looking at the effects of other variables on the fitted estimates. The authors illustrate this analysis by examining the effects of presentation duration on the parameter estimates produced by models that instantiate the 2 explanations. Although the question remains open, the storage based interpretation was found to be somewhat more plausible. PMID- 10789203 TI - Naive empiricism and dogmatism in confidence research: a critical examination of the hard-easy effect. AB - Two robust phenomena in research on confidence in one's general knowledge are the overconfidence phenomenon and the hard-easy effect. In this article, the authors propose that the hard-easy effect has been interpreted with insufficient attention to the scale-end effects, the linear dependency, and the regression effects in data and that the continued adherence to the idea of a "cognitive overconfidence bias" is mediated by selective attention to particular data sets. A quantitative review of studies with 2-alternative general knowledge items demonstrates that, contrary to widespread belief, there is (a) very little support for a cognitive-processing bias in these data; (b) a difference between representative and selected item samples that is not reducible to the difference in difficulty; and (c) near elimination of the hard-easy effect when there is control for scale-end effects and linear dependency. PMID- 10789204 TI - Seven (indeed, plus or minus two) and the detection of correlations. AB - Capacity limitations of working memory force people to rely on samples consisting of 7 +/- 2 items. The implications of these limitations for the early detection of correlations between binary variables were explored in a theoretical analysis of the sampling distribution of phi, the contingency coefficient. The analysis indicated that, for strong correlations (phi > .50), sample sizes of 7 +/- 2 are most likely to produce a sample correlation that is more extreme than that of the population. Another analysis then revealed that there is a similar cutoff point at which useful correlations (i.e., for which each variable is a valid predictor of the other) first outnumber correlations for which this is not the case. Capacity limitations are thus shown to maximize the chances for the early detection of strong and useful relations. PMID- 10789205 TI - [DDT and it metabolites in fish in the Gulf of Gdansk]. AB - The residues of DDT and its metabolites (DDTs; p,p'-DDT, o,p'-DDT, p,p'-DDD, o,p' DDD, p,p'-DDE, o,p'-DDE i p,p'-DDMU) has been determined in ten species of edible fish caught in the Gulf of Gdansk in 1992. The method of measurement was capillary gas chromatography and low resolution mass spectrometry (HRGC/LRMS) after a nondestructive extraction and clean-up step with a further fractionation of the extract on Florisil column. All fish examined contained detectable residues of DDTs, and the concentrations ranged from 28 to 310 ng/g wet weight. o,p'-DDT accounted from 0.4 to 2.5% to DDTs content. The residue concentration of DDTs in herring (110 ng/g wet weight and 1100 ng/g lipid weight) in 1992 was threefold lower than in the years 1979-1983 and fourteen fold lower than in 1969 1973. PMID- 10789206 TI - [Lead and cadmium content in daily food rations of children and adolescents from copper basin of the Legnica region]. AB - Environmental pollution still remains a serious problem in some heavily industrialized regions of Poland. Among various xenobiotics prevalent in human environment heavy metals such as lead and cadmium are considered as most harmful to living systems. The main source of these metals for humans is food and water, therefore increased dietary lead and cadmium intake may cause functional disturbances of various body systems, especially in young developing organisms. The objective of this study was to evaluate the lead and cadmium content in daily food rations served at two selected Children Guardian Centres (CGC) located in the region of Copper Basin Legnica (Glogow and Polkowice), during winter and spring seasons in 1996. The lead and cadmium content was determined after dry mineralization of samples by the FAAS method with deuterium BC. It was found that the lead and cadmium content in daily food rations exceeded the PTWI limits for children and adolescents in both CGC. Moreover, seasonal and regional variations of the lead and cadmium content in daily meals were observed. Generally, higher levels of lead in food rations were determined in the winter season whereas the cadmium content was higher in the CGC in Glogow in comparison to Polkowice. PMID- 10789207 TI - [The content of selected minerals (Ca, Mg, Zn, Cu, Fe) in daily food rations of adolescents: comparison of analytical and calculated data]. AB - The content of calcium, magnesium, zinc, copper and iron in daily food rations of adolescents served in two residential centres in Copper Basin Legnica region and two high schools in Poznan were studied. The content of minerals was determined by two independent methods: chemical analysis and calculation based upon food composition tables. It was found that the calculated results were markedly higher, suggesting that daily meals covered recommended allowances for minerals established for the adolescents group. However, the analytical results were 10% lower on average, but in the case of zinc and magnesium they did not even reach 50% of norms showing that the daily food did not provide sufficient amounts of minerals for adolescents. Moreover, analysis of regression showed a significant correlation for the magnesium, iron, zinc and copper content obtained by the two methods which allowed to calculate empirical formulae that may help to establish the content of these elements in daily food with the use of the FOOD computer program. PMID- 10789208 TI - [Fatty acids in chocolate and chocolate products]. AB - Fat content as well as fatty acids composition in 46 chocolate both stuffed and hard type as well as 14 chocolate related products from the market in Warsaw area in the years of 1997-1999 has been analysed. As the result of above investigations the considerable difference in fat content (from 6.6 to 40.0%) as well as fatty acids groups has been detected, in particular in stuffed chocolates and chocolate products. PMID- 10789209 TI - [The content of nitrates and nitrites in dietary rations of students of Gdynia Maritime Academy]. AB - The content of nitrates (V) and (III) in dietary rations of Gdynia Maritime Academy students was determined. A variable content of examined compounds was detected. Average content of nitrates (V) was 224.92 mg NaNO3/person/day, whereas of nitrates (III) 2.14 mg NaNO2/person/day. PMID- 10789210 TI - [Dietary intake of dieldrin and aldrin in Poland]. AB - Dieldrin and aldrin exposure from a particular food items in Poland in 1970-1996 was calculated by multiplying its annualized mean consumption rates by residue concentration in the food. Estimated daily dietary intakes of dieldrin and aldrin were from 1.0 do 1.3 micrograms per person in 1970-1985 and from 0.50 do 0.58 microgram per person in 1990-1996, on the average. Fish and dairy products are a main source of dieldrin in a total diet in Poland. PMID- 10789211 TI - [Evaluation of the biological action of the fluoride waters applied in the form of the potable cure in experimental animals]. AB - Basing on the carried out investigations it has been shown that the fluoride water from the spring "Zdzislaw" from Ladek Zdroj and the aqueous solution of NaF in the F concentration like that in the curative water, used in rats intragastrically in a single daily dose of 14.3 ml/kg of body weight or ad libitum for a period of 20 days, brought about a statistically significant decrease of whole cholesterol, of the HDL fraction of cholesterol, of whole lipids, triglycerides, sodium and magnesium and about an increase of potassium and calcium in the blood serum as compared with the control group of animals. Observed at the same time were changes that testified to the occurrence of respiratory decompensated acidosis and alkalosis. No essential influence of the investigated waters on the protein, carbohydrate metabolism, smear and the morphological composition of the peripheral blood and on the bone marrow smear could be stated. The investigated waters acted spastically on the smooth muscles of the small intestine of the rabbit, they were showing diuretic action in the rats and caused an increased water turnover in the organism. PMID- 10789212 TI - [Determination of vitamins B1 and B2 in selected vegetable-fruit products]. AB - A simple method was described to determine vitamins B1 and B2 in fruit and vegetable-fruit juices. Vitamins were extracted with perchloric acid and for their determination an ion pairing RP-HPLC technique with UV detection by 254 nm was applied. The average recovery for vitamin B1 was 85-104% and 94-105% for vitamin B2. The statistically estimated limits of identification and detection were respectively, for both vitamins: for B1: 0.008 and 0.0024, and for B2 0.0002 and 0.0007 mg/ml of product. PMID- 10789213 TI - [Modification of enzymatic method of cholesterol determination]. AB - Fluorimetric substrates for peroxidase activity determining were described in this work so that to establish cholesterol concentration by enzymatic method. p hydroxyphenyloacetate acid was used, as the most favourable, so that to determine cholesterol content in meat products. Using of this compound enables to diminish cholesterol determining limit more than 10-times. Obtained results of cholesterol content are convergent with results obtained by the other authors. PMID- 10789215 TI - [Evaluation of a diet and nutritional status of breast feeding women on the basis of a questionnaire study]. AB - The subjects were 120 breast-feeding mothers whose children were admitted for outpatient or inpatient treatment at the Institute of Mother and Child in Warsaw, between March, 1998 and July, 1999. A special nutritional questionnaire was elaborated with the aim to evaluate subjects' nutritional status as well as their diet. All subjects completed the questionnaire and then selected parameters of their nutritional status and diet were evaluated. For 43 of the women under study it was the first month of lactation, for 33--the fourth month, for 23--the sixth and 21 women had been breast-feeding for more then six months. The subjects were of different age, coming from both urban and rural areas. In the studied group the nutritional status, i.e. body mass indices (BMI) before pregnancy, at the end of pregnancy and during lactation were within the normal range. 65 of the subjects had changed their diet due to pregnancy and 52 due to lactation. Women declared to be used to eating 3-4 meals per day. Having additional snacks was declared by 62.5%. Liquids' intake was on average about 2.5 l/24 hr, range 2-31. Average daily energy intake--2938 kcal/12,980 kJ, the lowest--in a diet of a mother with diabetes, the highest--in a diet of a vegetarian mother. The proportion of protein in energy intake was 15.1%, whereas fat--36.3%. Dietary practices of women under study in the period of lactation were well adjusted to nutritional recommendations for this group, as evidenced by indices of nutritional status (i.e. BMI). Among 120 women, 71 were taking supplementary preparations of vitamins and minerals. It turned out that a level of some vitamins in a daily food intake together with supplementation was definitely too high. PMID- 10789214 TI - [New food product containing polyunsaturated fatty acids Omega-3 EPA, DHA: sensory quality and possibility of diet supplementation]. AB - Polyunsaturated fatty acids omega-3, especially long chain forms--EPA, DHA, exert desirable influence on human health by decreasing risk of many life-style diseases. However intake of these is usually too low, that brings many unfavourable health effects. Increased consumption of omega-3 fatty acids (up than 200 mg/day) is now commonly recommended. In Poland EPA, DHA intake is only about 100 mg/day, one of the lowest in Europe. A way to arise EPA, DHA amount in the diet, without radical changes of eating habits, seems to be consumption of foods containing its addition. The aim of the study was to explore the possibility of omega-3 polyunsaturates EPA, DHA in microencapsulated form use for formulation of high sensory quality pastilline form foods (orange and mint flavour versions), and to evaluate influence of 4 mts. storage time (room temp., no access to light and air) on sensory quality. Sensory evaluation was performed using methods of sensory analysis. Following sensory features were evaluated: palatability (structurized graphic scale method), intensity of fishy flavour (non structurized graphic scale method), and sensory quality (succesion method). Results were statistically interpreted. It was shown that palatability of elaborated foods wasn't significantly affected by EPA, DHA concentration up to 0.8-1%, depending on presence of flavours masking fishy taste of omega-3 polyunsaturates, during whole storage time. Daily intake of 5-10 units of elaborated products provides 65-150 mg EPA, DHA, i.e. 35-70% of its minimal proposed intake (total energy 22-44 kcal, i.e. 91-182 kJ). That may increase EPA, DHA amount in average Polish diet by 70-140%, elevating them towards recommended one. PMID- 10789216 TI - [Changes of body fat mass determined by be-electrical impedance and by anthropometry: BMI method and skinfolds method in overweight and obese women after implementation of low energy diet]. AB - OBJECTIVE: To estimate the effect of a low energy diet on body fat mass measured by bioelectrical impedance (BIA) in overweight (BMI > or = 25 < 30) and obese women (BMI > or = 30). DESIGN: Randomised six weeks trial. METHODS: The studied group consisted of 86 women aged 20-67 attended the Outpatients Clinic of Metabolic Disorders. Dietary assessments were performed by 24-hour dietary recall. Body fat mass was measured by bioelectrical impedance method and by skinfolds thickness method. Percent body fat and fat free mass was estimated from equations based on BMI and skinfolds thickness. RESULTS: In overweight women after 6 weeks of dietary treatment mean intake of energy decreased from 10071 +/- 2678 kJ to 4560.6 +/- 1405.8 kJ, total fat intake from 88.7 +/- 33.9 g/d (33.4% of energy) to 38.8 +/- 19.9 g/d (31.1% of energy), protein intake from 89.5 +/- 36.5 g/d (14.8% of energy) to 50.7 +/- 16.9 g/d (19.0% of energy), carbohydrates intake from 312.7 +/- 106.6 g/d (51.8% of energy) to 134.5 +/- 53.7 g/d (50% of energy). In obese women mean intake of energy decreased from 10,376.3 +/- 2953.9 kJ to 4665.2 +/- 1380.7 kJ. The value of total energy, total fat and saturated fatty acids intake correlated with body weight, BMI and body fat. After dietary treatment the body weight decreased by 2.4 kg (3.3%) in overweight women and by 3.9 kg (4.1%) in obese women and percent of body fat decreased by 1.6% and 2.3%, respectively. Body fat mass determined by BIA method significantly correlated with skinfolds method by Siri, Schutte, Rathbun, Brozek, Keys-Brozek, and BMI method by Webster, Deurenberg. CONCLUSION: BIA method may be a helpful tool for the analysis of changes in total body composition occurring under obesity treatment. PMID- 10789217 TI - [The state of insurance medicine in Russia]. PMID- 10789219 TI - [The role of radiotherapy in early breast cancer]. PMID- 10789218 TI - [Adjuvant therapy in breast cancer]. PMID- 10789220 TI - [Chemotherapy in the combined treatment of cervical cancer]. PMID- 10789221 TI - [Registration of patients with multiple primary malignant neoplasms]. PMID- 10789222 TI - [Long-term results of hyperfractionated radiotherapy in patients with stage III IV oropharyngeal carcinoma]. AB - Radiotherapy was given to 139 patients with oropharyngeal carcinoma (stage III IV) divided into two groups. Group I included 82 patients who received three daily equal fractions (1 Gy), at 4 hr interval, of a mean total focal dose of 80 Gy. Patients in group II (57) received a mean total dose of 68 Gy in 5 weekly standard fractions. Fractionated treatment was carried out in both groups in three stages at 17-day intervals. Complete remission was achieved in 50 (60.9%) patients in group I and in 31 (54.4%)--group II. Total 5-year and recurrence-free survival was 37.3 +/- 5.4 and 32.4 +/- 5.2% (group I) and 12.5 +/- 4.7 and 10.4 +/- 4.4% in group II, respectively. Radiation injury rates were nearly identical in both groups: 14% for standard radiotherapy and 12% for multiple fractions. The data point to high effectiveness of the latter. PMID- 10789223 TI - [Fatty acid composition of lipids in lung tissue of patients with lung cancer]. AB - The fatty acid (FA) spectrum of lung tissue fragments was studied versus distance of malignant cells from lesion focus in lung cancer patients. Decreased polyunsaturated FA levels in all samples were attributed, chiefly, to linoleic, (linoleic-new) and eicosatrienoic acid deficiency. Both in tumor tissue and areas of intensive cell proliferation, accumulation of monounsaturated C18:1 and C15:1 fatty acids, low C20:4(6 and C 20:5(3 and an FA with 19 carbon atoms were detected. The total concentration of (-6 and (-3 series fatty acids was relatively low, too, the deficit of the latter being remarkably low. Those changes are likely to be related to certain peculiarities of the structure and functions of tumor cells. PMID- 10789224 TI - [Parameters of lipid metabolism and polymorphism of apolipoprotein aI and angiotensin-converting enzyme genes in patients with endometrial carcinoma]. AB - Data on lipid metabolism in 109 cases of endometrial carcinoma and 33 patients in control are presented. Relevant disorders were detected in 72.5% of the study group [stage I obesity--23 (21.1%); II--29 (26.6%); III--25 (22.9%); IV--2 (1.9%)] which was 1.5 times the mean level in controls (51.5%). The abdominal pattern of obesity was predominant (77.7%). Symptoms of cardiosclerosis were identified in 93 patients with endometrial tumors (85.3%), 93.5% of the latter group presenting with concomitant hyperglyceridemia. In 68.8% of endometrial carcinoma patients, incidence of arterial hypertension was higher than in controls (54.5%). Lipoproteins played a major role in dyslipoproteid pathogenesis involved in obesity and high blood pressure. A study of the insertion-deletion (I/D) polymorphism of apolipoprotein AI genes showed two deletion alleles (DD- 6%) and one heterozygote (ID--3%) in control group; no deletion alleles were identified in endometrial tumor patients (0.1 (p(0.05). An investigation of the I/D polymorphism of angiotensin-converting enzyme genes identified deletion homozygotes in 30, insertion homozygotes (II)--27, and heterozygotes--41% (control). In endometrial cancer group, the deletion allele distribution was: DD- 29; II--28 and ID--45%. Deletion allele frequency in control was 0.485 while in endometrial carcinoma--0.484 (p(0.05), i.e. with out significant difference. PMID- 10789225 TI - [Conformational changes in C3 levels of complement during storage of blood from Hodgkin's disease patients and from healthy people]. AB - Storage-related differences in C3 level of blood complement and its hydrolized form--C3(H2O)--were identified in Hodgkin's disease patients and healthy donors, by immunoenzymatic analysis using murine monoclonal antibodies. Both C3 and C3(H2O) levels in blood serum of patients varied with time and were significantly different from those in health subjects; they correlated with EDTA concentration. After a second thawing of plasma in patients, there were no traces left of C3(H2O). PMID- 10789226 TI - [Autoimmune reactions in patients with various tumor localizations]. AB - Autoantibody levels in patients with malignancies of the oral cavity and lung and autoimmune disorders have been assayed. It was found that tumors initiated polymorphous autoimmune reactions to various tissue antigens. Autoantibody production rates in tumor patients were comparable with those in autoimmune disorders. The role of autoimmune reactions in cancer pathogenesis is discussed. PMID- 10789227 TI - [Exposure to chloroprene as an occupational hazard (a cohort epidemiologic study)]. AB - A cancer epidemiological study was conducted in chloroprene workers in Yerevan. In a cohort of male workers, the total morbidity rates for all tumor patterns were significantly lower (32%) than expected, which was attributed to the effect of "the healthy worker". Risk for hepatic tumor was 3.83 times the expected level among male operators of the chloroprene production equipment. A direct "dose effective" correlation was established between liver cancer risk and length of record for occupational exposure to chloroprene. The highest risk was among males who came into contact with the substance under the age of 30. PMID- 10789228 TI - [Inhibitory effect of progesterone P1-1 on glutathione-s-transferase and its antiproliferative effect on human erythroleukemia K562 cells]. AB - Since glutathione S-transferase P1-1 (GST P1-1) is predominantly expressed in many types of tumor cells it is regarded as its marker protein. The report discusses data on a relationship between progesterone (PS) inhibition of GST P1-1 and proliferation of human erythroleukemia K562 cells. Unlike such steroids as estrone, dexamethasone, testosterone and hydrocortisone, PS showed significant inhibitory effect (I50 = 32.5(M) and noncompetitively (Ki = 25(M) inhibited GST P1-1 isolated from human placenta. Pronounced inhibition of K562 cell proliferation by PS was in inverse correlation with the intracellular activity of GST P1-1. Also, PS injected into culture suppressed GST P1-1 expression in leukemia cells. At concentrations of 10(-8)-10(-5) M, both effects proved dose dependent. The correlation between PS antiproliferative effect on GST P1-1 activity and expression in K562 cells is discussed in terms of its possible role played in malignant growth regulation. PMID- 10789230 TI - [Role of laparoscopy in the diagnosis and treatment of ovarian cancer]. AB - Diagnostic potential of laparoscopy and treatment of ovarian cancer were evaluated using the findings of 322 examinations: (1) 248 patients with sonography-detected masses, 2-7 cm, in the small pelvis; (2) 54 cases of morphologically inconclusive data on clinically advanced tumor, and (3) 50 patients with ovarian carcinoma in clinical remission, following complex treatment. Owing to its high predictive value, laparoscopic surgery should be recommended for comprehensive diagnosis of small-size masses located in the small pelvis as possible early precursors of ovarian tumor; morphological verification of tumor process extent, and detection of preclinical recurrence of tumor in cases of failed non-invasive monitoring procedures such as ultrasound and CA-125 determination. PMID- 10789229 TI - [Surgical treatment of stage IIIA non-small-cell lung cancer]. AB - Data on five-year survival were evaluated for 258 patients with non-small cell lung cancer (stage IIIA) (N2). In 155 patients (60%), N2 tumor was detected during surgery. Total resection was carried out in 179 (69.4%), subtotal--79 (30.6%). Total lymph node dissection was not employed in the latter group. Lateral thoracotomy was used in 213 cases. Transsternal procedure was performed in 45 cases of bulky tumor and extensive invasion of mediastinal fat. A comparison of five-year survival data failed to establish any relationship between survival and postoperative radiochemotherapy in radically-operated patients. It was found that surgery for non-small lung N2 tumors with mediastinal involvement is indicated and may be effective if total lymph node dissection is performed. PMID- 10789231 TI - [Experience with conservative surgery for bone sarcoma involving major neurovascular bundles]. PMID- 10789232 TI - [Diagnostic potential of ultrasound examination for tumor recurrence and metastasis]. AB - Ultrasound examination was carried out in 252 patients following specific treatment of uterine and ovarian malignancies. To compare the effectiveness of diagnostic procedures, transabdominal and transvaginal examinations were performed in succession in all the patients using ultrasound (Doppler) technique. Transvaginal study proved relatively more sensitive (98.9%) and diagnostically accurate (97.2%) in detecting relapse. Transabdominal examination carried out in conjunction with color Doppler imaging was best for metastasis detection. PMID- 10789233 TI - [Experience with recombinant erythropoietin for anemia in children with malignant tumors]. PMID- 10789235 TI - [Psychologic aspects of the physician's work]. PMID- 10789234 TI - [A case of difficult morphologic diagnosis of Ewing's sarcoma in a child]. PMID- 10789236 TI - [Thoracoscopy in current oncology]. PMID- 10789237 TI - [Cytoreductive surgery in the treatment of stage-IV colorectal cancer--is it justified?]. PMID- 10789238 TI - Living longer. PMID- 10789239 TI - Living longer. PMID- 10789241 TI - Fireballs of free quarks PMID- 10789240 TI - Dissecting the mind. PMID- 10789242 TI - Outbreak not contained. West Nile virus triggers a reevaluation of public health surveillance. PMID- 10789243 TI - A taste for science. PMID- 10789245 TI - Power to the PC. PMID- 10789244 TI - Car parts from chickens. Researchers hatch a plan to make plastic from feathers. PMID- 10789246 TI - Rules of the game. PMID- 10789247 TI - Quantum teleportation. PMID- 10789248 TI - Building a brainier mouse. PMID- 10789249 TI - Understanding clinical trials. PMID- 10789250 TI - The discovery of brown dwarfs PMID- 10789251 TI - The Aleutian kayak. PMID- 10789252 TI - Monitoring earth's vital signs. PMID- 10789253 TI - Who were the Neandertals? PMID- 10789254 TI - Cleaning agents. PMID- 10789255 TI - A furnace in a thermos. PMID- 10789256 TI - Counting the cattle of the sun. PMID- 10789257 TI - Massive obstetric haemorrhage. AB - Massive obstetric haemorrhage is a major cause of maternal death and morbidity; abruptio placentae, placenta praevia and postpartum haemorrhage being the main causes. A delay in the correction of hypovolaemia, a delay in the diagnosis and treatment of defective coagulation and a delay in the surgical control of bleeding are the avoidable factors in most maternal deaths caused by haemorrhage. The degree of hypotension is the first guide to the level of blood loss, except in abruptio placentae. A protocol incorporating the guidelines is shown. The rapid correction of hypovolaemia with crystalloids and red cells is the first priority, followed by blood component therapy as indicated by the haematocrit, coagulation tests, platelet count and clinical features. Serial monitoring of the response to treatment is essential. Oxytocin and prostaglandin will correct uterine atony, and appropriate surgical intervention is required for traumatic bleeding. Ligation of the uterine arteries, ovarian arteries and internal iliac arteries will usually control uterine bleeding, arterial embolization also being effective. Hysterectomy should be considered as well. Catastrophic bleeding may also arise in complications such as rupture of the liver and acute fatty liver of pregnancy. These rare complications are best managed by a multidisciplinary team involving the obstetrician, anaesthetist, haematologist, hepatologist and renal physician. The rupture of aneurysms in the splenic artery and in other branches of the aorta can result in massive haemorrhage during pregnancy and following delivery. PMID- 10789258 TI - Non-haemorrhagic obstetric shock. AB - The causes of non-haemorrhagic obstetric shock (pulmonary thromboembolism, amniotic fluid embolism, acute uterine inversion and sepsis) are uncommon but responsible for the majority of maternal deaths in the developed world. Clinically suspected pulmonary thromboembolism should be treated initially with heparin and objective testing should be performed. If the diagnosis is confirmed, heparin is usually continued until delivery, following which anticoagulation in the puerperium is achieved with either warfarin or heparin. Amniotic fluid embolism is a rare complication of pregnancy, occurring most commonly during labour. The management of amniotic fluid embolism involves maternal oxygenation, the maintenance of cardiac output and blood pressure, and the management of any associated coagulopathy. Acute uterine inversion arises most commonly following mismanagement of the third stage of labour. The shock in uterine inversion is neurogenic in origin, although there may also be profound haemorrhage. The management of this condition includes maternal resuscitation and replacement of the uterus either manually, surgically or by hydrostatic pressure. Genital tract sepsis remains a significant cause of maternal death, the most common predisposing factor being prolonged rupture of the fetal membranes. The management of septic shock in pregnancy includes resuscitation, identification of the source of infection and alteration of the systemic inflammatory response. PMID- 10789259 TI - Emergencies in operative obstetrics. AB - Among all the emergency situations which may arise across the field of obstetrics and gynaecology, there are a small number which call for urgent practical steps to be taken in order to safeguard the life of the mother or the baby or both. The three such complications dealt with in this chapter consist of one prior to delivery--prolapse of the umbilical cord; one during delivery--shoulder dystocia; one following delivery--acute inversion of the uterus. All of the above require prompt action by well-trained staff and may involve the active and efficient co operation of a range of different health care professionals. It is critically important that staff are fully aware of the procedures to be followed and the chain of command which will ensure that they are followed as efficiently and successfully as possible. PMID- 10789260 TI - Severe pre-eclampsia and eclampsia. AB - The mainstay of the management of severe pre-eclampsia is early referral, stabilization of the mother with antihypertensive therapy and anticonvulsants if required, full assessment of the mother and the baby, and delivery on the best day in the best way. It is to be remembered that delivery is the long-term cure, but most women get worse after delivery and most maternal deaths occur postpartum. It is important that doctors have the training to be aware of the dangers of this condition, guidelines to follow and senior support. Lowering blood pressure has been associated with a reduction in the mortality from cerebrovascular accident and early use of antihypertensive agents is beneficial to both mother and baby. The main cause of death is now pulmonary oedema, with renal failure a rare complication. It is important that, after delivery, vigilance is maintained and fluid replacement is given with care. It is better to 'run them dry' than to give fluid replacement that may encourage pulmonary oedema. Followup is required with counselling about what has happened and the prospects of recurrence. PMID- 10789261 TI - Acute psychiatric disturbance in pregnancy and the puerperium. AB - Psychiatric disturbance occurring during pregnancy or the puerperium can occur as a first presentation or as a recurrence or deterioration of a previously diagnosed psychiatric illness. Whatever the history or aetiology, the occurrence of psychiatric disturbance during this period can have profound, long-term implications not only for the mental health of the mother, but also for her relationship with her child and other family members, as well as for the child's development. The treatment of psychiatric disturbance at this time can present difficult dilemmas and should always involve a careful analysis of the risks and benefits of treatment. Liaison between obstetricians and psychiatrists is to be strongly encouraged in order to ensure the best possible outcome for both mother and baby. The antenatal period offers a unique opportunity to identify individuals at risk and target them in order to promote psychological well-being during pregnancy and the puerperium. PMID- 10789263 TI - Ovarian hyperstimulation syndrome. AB - Ovarian hyperstimulation syndrome is an iatrogenic complication of assisted reproduction. In its most severe form, it is potentially fatal. The major clinical components are marked ovarian enlargement and increased capillary permeability leading to ascites, hydrothorax and pericardial effusion. Severe cases are associated with thromboembolic phenomena, respiratory distress and renal failure. The definitive pathophysiology is unknown. The available evidence would support a central role for inflammatory cytokines and angiogenic growth factors. Ultrasound examination and serum oestradiol values are currently used to predict patients at risk. The ideal treatment is prevention, but there has been only limited success. The main aims of treatment are to correct fluid imbalance, maintain renal perfusion and support the patient until the condition resolves. Drug therapy has a limited role, although anticytokine agents may prove useful. PMID- 10789262 TI - The acute abdomen and the obstetrician. AB - An acute abdomen in pregnancy can be caused by pregnancy itself, be predisposed to by pregnancy or be the result of a purely incidental cause. These various conditions are discussed. The obstetrician often has a difficult task in diagnosing and managing the acute abdomen in pregnancy. The clinical evaluation is generally confounded by the various anatomical and physiological changes occurring in pregnancy itself. Clinical examination is further hampered by the gravid uterus. The general reluctance to use conventional X-rays because of the pregnancy should be set aside when faced with the seriously ill mother. A reluctance to operate during pregnancy adds unnecessary delay, which increases morbidity for both mother and fetus. Such mistakes should be avoided as prompt diagnosis and appropriate therapy are crucial. A general approach to acute abdominal conditions in pregnancy is to manage these problems regardless of the pregnancy. PMID- 10789264 TI - Post-operative complications following minimal access surgery. AB - The growth in popularity of minimal access surgery in gynaecology has coincided with a reduction in the length of hospital stay. Patients need to be made aware of the potential for and symptoms and signs of complications to ensure that they seek help at an early stage. Clinicians need to be vigilant in the recognition of adverse events both intra- and post-operatively to minimize the risk of developing serious complications. The growth in litigation in minimal access surgery represents, in part, a failure of surgeons to adapt to the new demands of this type of surgery. PMID- 10789265 TI - Preterm labour. AB - Spontaneous preterm labour remains a major obstetric problem because of the high incidence of neonatal mortality or long-term handicap associated with it. The drugs available for the management of preterm labour are poorly effective and have potentially serious side-effects for the mother or fetus. In recent years, there has been a remarkable increase in the knowledge of the biochemical mechanism underlying uterine quiescence and contractility. Many of the G protein coupled receptors that participate in the regulation of myometrial activity have been cloned and characterized, and their intracellular signalling pathways have been elucidated. The role of G protein receptor kinases in uterine tachyphylaxis is better understood. New developments in our understanding of the cellular mechanisms involved in uterine contractions in idiopathic and infection associated preterm labour are expected, which will lead to better, more selective therapy for this problem. However, much research remains to be done before the mechanism of human parturition is fully understood. PMID- 10789266 TI - Fetal distress. AB - The term 'fetal distress' should be replaced by 'suspected fetal compromise' because the diagnosis of 'fetal distress' is often unproven. Cardiotocography remains the cornerstone of making the diagnosis, but as a test it is renowned for its high sensitivity and low specificity. It has reduced intrapartum fetal mortality but not long-term neonatal morbidity or the incidence of cerebral palsy. There is no doubt that when obvious signs of fetal compromise, such as late decelerations in the presence of intrauterine growth retardation and oligohydramnios, are present, the diagnosis of fetal compromise is relatively simple. Often, however, the subtle signs of fetal compromise are missed; these are a change in the grade of meconium in the amniotic fluid, a rising base-line fetal heart rate, the absence of accelerations, the presence of 'atypical' variable decelerations or a combination of the above. To date, there is no test available to replace the cardiotocograph, although fetal pulse oximetry is the most promising adjunctive test. Above all, no test result obtained in isolation must detract from the whole clinical picture. PMID- 10789268 TI - How much English health authorities are allocated for mental health care. PMID- 10789267 TI - Twin studies of psychosis and the genetics of cerebral asymmetry. PMID- 10789269 TI - Greater occurrence of schizophrenia in dizygotic but not monozygotic twins. Register-based study. AB - BACKGROUND: A previous study demonstrated a higher rate of first hospitalisation for schizophrenia in twins than in singletons. AIMS: To compare the schizophrenia rates in monozygotic twins, dizygotic twins and the general population. METHOD: Through record linkage between the Young Cohort of the Danish Twin Register and the Danish Psychiatric Case Register, the rate of schizophrenia in monozygotic and dizygotic twins and the rate in the general population were compared. RESULTS: The rate of first admission to hospital for schizophrenia in dizygotic twins was 40% greater than that in the general population. The rate in monozygotic twins was not increased. The difference between the rates in mono- and dizygotic twins was statistically significant (P < 0.001). CONCLUSIONS: This result is surprising and is in the direction opposite to that expected, suggesting new possibilities in the study of risk factors. PMID- 10789270 TI - Schizophrenia with onset after age 50 years. I: Phenomenology and risk factors. AB - BACKGROUND: Schizophrenia occurring for the first time in late life may be a distinct entity or part of a continuum. AIMS: Can late-onset schizophrenia (LOS) and early-onset schizophrenia (EOS) be differentiated by their phenomenology and risk factors to their development? METHOD: Convenience samples of 27 DSM-III-R defined LOS subjects, 30 EOS subjects and 34 control subjects were systematically assessed. RESULTS: Premorbidly, both groups of subjects with schizophrenia had personality traits that were different from controls but not from each other. The EOS subjects had more family members with a history of psychiatric illness or schizophrenia and less hearing impairment than the other two groups, which did not differ from each other. Clinically, LOS and EOS subjects were similar, except that EOS subjects had more negative symptom scores, tended to have more delusions of guilt/sin and of being controlled and more formal thought disorder, and had significantly poorer instrumental activities of daily living. CONCLUSIONS: Phenomenology and risk factors do not distinguish discrete LOS. PMID- 10789271 TI - Schizophrenia with onset after age 50 years. 2: Neurological, neuropsychological and MRI investigation. AB - BACKGROUND: Late-onset schizophrenia (LOS) may have a basis in age-related coarse brain disease, but empirical support for this is conflicting. AIMS: Is LOS characterised by more neurological disease than early-onset schizophrenia (EOS)? METHODS: DSM-III-R-defined LOS subjects (n = 27) were compared with 30 EOS and 34 volunteer control subjects on neurological status, neuropsychological test performance and brain magnetic resonance imaging (MRI). RESULTS: LOS and EOS groups had more 'soft' neurological signs and drug-induced movement abnormalities, and performed more poorly on tests assessing speed of information processing, memory and frontal executive functioning. On MRI, the LOS and EOS groups had greater lateral ventricular size than the control group. The LOS subjects also had more signal hyperintensities in periventricular white matter and subcortical nuclei than controls. CONCLUSIONS: LOS and EOS subjects were very similar on neuropsychological, neurological and structural neuroimaging parameters, except that there were more MRI periventricular hyperintensities in LOS subjects. PMID- 10789272 TI - Operation and organisation of services for people with severe mental illness in the UK. A survey of the Care Programme Approach. AB - BACKGROUND: Since 1991, English mental health policy guidelines have been explicit in recommending inter-professional working, involvement of patients and carers, harmonisation between health and social services and targeting at people with severe mental illness. AIMS: To explore the structure and operation of mental health services in practice and relate them to measurable data. METHOD: A survey of mental health trusts was conducted in 1997-1998. Responses were compared with a concurrent survey of social services departments. Some qualitative data were collected. Harmonisation and targeting scores were devised and tested for association. RESULTS: The response rate was 79%. The frequency with which different professionals, patients and carers were involved in the care process is described. Reasons for this are discussed briefly. Levels of harmonisation and targeting were found to vary widely and to be positively associated. CONCLUSIONS: This survey portrays mental health services' practice in relation to Government policy. However, adherence to guidelines cannot be taken as a guarantee of service quality or efficiency. PMID- 10789273 TI - Predictors of quality of life in people with severe mental illness. Study methodology with baseline analysis in the UK700 trial. AB - BACKGROUND: It is not clear which model of case management is most likely to improve quality of life in people with severe mental illness. AIMS: To use baseline data derived from the UK700 Case Management Trial to assess the relative importance of clinical, social and unmet needs variables as predictors of subjective quality of life in patients with severe mental illness. METHOD: Patients (n = 708) were assessed on quality of life (Lancashire Quality of Life Profile), needs (Camberwell Assessment of Need), psychopathology and social functioning. Variables that were amenable to change through case management were investigated as predictors of quality of life. RESULTS: Social variables accounted for 7% of the variance for subjective quality of life, compared with 19% for clinical variables, and 20% for unmet needs. The strongest predictors of subjective quality of life were unmet basic, social and functioning needs, depression and positive psychotic symptoms. CONCLUSIONS: Subjective quality of life in severely mentally ill patients is predicted by clinical variables and unmet needs. The results identify priority areas for the attention of case managers who seek to improve quality of life in these patients. PMID- 10789274 TI - Undifferentiated dementia, Alzheimer's disease and vascular dementia: age- and gender-related incidence in Liverpool. The MRC-ALPHA Study. AB - BACKGROUND: Does incidence of dementia follow the age pattern of prevalence? Is gender a risk factor? Do patterns of incidence differ between dementias? AIMS: To assess age-specific incidence rates of undifferentiated dementias, Alzheimer's disease and vascular dementia. METHOD: 5222 individuals aged > or = 65 years, were interviewed using the Geriatric Mental State/History and Aetiology Schedule. The AGECAT package was used to identify cases at three interviewing waves at two year intervals. Diagnoses were made using ICD-10 Research Criteria and validated against neurological and psychological examination, with imaging and neuropathology on unselected subsamples. RESULTS: Incidence rates of the dementias increase with age. Age patterns are similar between Alzheimer's disease and vascular dementia. Gender appears influential in Alzheimer's disease. In England and Wales, 39,437 new cases of Alzheimer's disease (4.9/1000 person-years at risk); 20,513 of vascular dementia (2.6/1000 person-years) and 155,169 of undifferentiated dementia (19/1000 person-years) can be expected each year. CONCLUSIONS: Incidence rates for Alzheimer's disease and vascular dementia appear to behave differently, with an increased risk of Alzheimer's disease for women compared to vascular dementia. PMID- 10789275 TI - Natural history of pharmacotherapy of older depressed community residents. The MRC-ALPHA Study. AB - BACKGROUND: Depression in older people is common and has a high mortality, but effective treatments exist. AIMS: To describe drug prescribing in older community residents in relation to depression status. METHOD: The MRC-ALPHA community cohort aged 65 and over were interviewed using the Geriatric Mental State examination drug data collected at index interview and at two and four years. RESULTS: Antidepressants were used by 10.9% of the depressed population. Benzodiazepines were used frequently. Of the antidepressant users, 59.6% took low dose antidepressants for two years, had a poor outcome and few drug changes. CONCLUSIONS: Trends of increasing antidepressant use have cost implications for primary care groups. Benzodiazepines may be mis-prescribed for treatment of depressive symptoms. Antidepressant users have poor outcome and follow-up. PMID- 10789276 TI - Autism screening questionnaire: diagnostic validity. AB - BACKGROUND: Good interview and diagnostic measures for autism and other pervasive developmental disorders (PDDs) are available but there is a lack of a good screening questionnaire. AIMS: To develop and test a screening questionnaire based on items in the best available diagnostic interview--the Autism Diagnostic Interview--Revised (ADI-R). METHOD: A 40-item scale, the Autism Screening Questionnaire (ASQ), was developed and tested on a sample of 160 individuals with PDD and 40 with non-PDD diagnoses. RESULTS: The ASQ has good discriminative validity with respect to the separation of PDD from non-PDD diagnoses at all IQ levels, with a cut-off of 15 proving most effective. The differentiation between autism and other varieties of PDD was weaker. CONCLUSIONS: The ASQ is an effective screening questionnaire for PDD. PMID- 10789277 TI - Validation of the Hospital Anxiety and Depression Scale for use with adolescents. AB - BACKGROUND: The Hospital Anxiety and Depression Scale (HADS), developed for use with adults, has potential with adolescents. AIMS: To test the HADS's validity with adolescents. METHOD: The HADS was given to 248 schoolchildren and to 48 psychiatric out-patients and 38 deliberate self-harm in-patients aged 12-17, and validated against ICD-10 diagnoses for the out-patients. RESULTS: The HADS has adequate test-retest reliability and factor structure, and discriminates between adolescents diagnosed with depressive or anxiety disorders and those without these diagnoses. CONCLUSIONS: The psychometric properties of the HADS and its shortness make it useful for screening and in clinical settings with adolescents. PMID- 10789278 TI - Conflict between mothers with eating disorders and their infants during mealtimes. AB - BACKGROUND: In a recent study, 20% of the variance in the weight of infants of mothers with eating disorders was accounted for by mealtime conflict. AIMS: To investigate the antecedents and interactive processes involved in the development of such conflict. METHOD: Mothers with eating disorders and their 12-month-old infants (n = 34) and a comparison group (n = 24) were videotaped during infant mealtimes. Specific classes of antecedents to conflict episodes were identified. An examination was then made of all such antecedents not leading to conflict and the mother-infant responses to each antecedent. RESULTS: Within the index group, conflict was less likely when mothers acknowledged infants' cues and were able to put aside their own concerns. The relationship between maternal responses and the evolution of conflictual interaction was confirmed in multiple regression analyses including both index and comparison groups. CONCLUSIONS: Mothers' and infants' responses to potential antecedents to conflict had an impact on whether mealtime conflict ensued. Conflict arose because maternal eating disorder psychopathology interfered with aspects of responsive parenting. PMID- 10789279 TI - Violence, homicide and suicide: strong correlation and wide variation across districts. AB - BACKGROUND: The clinical assessment and management of the risk of violence and suicide by people with mental illness may have to focus on environmental as well as individual factors. AIMS: To investigate possible associations between violence, homicide and suicide rates, population density and indices of deprivation, with particular reference to inner-city boroughs. METHOD: Coroners' statistics in London for homicide and suicide were obtained, with police-reported homicide and violence rates as a validity check. Correlations were made between these data and population density, the Jarman under-privileged areas score, and the Mental Illness Needs Index for each of the 32 London boroughs. RESULTS: Homicide rates had a 14.3-fold range, suicide a 4.4-fold range and interpersonal violence a 6.6-fold range. The variables under study were strongly correlated with each other. Rates were highest in boroughs with high population density and deprivation scores. The associations persisted when covarying for deprivation, age structure or population density. CONCLUSIONS: Because violence, homicide and suicide are so closely correlated, they are likely to be valid indices of the differences between adjacent boroughs; this has implications for the delivery of preventive and mental health services and for clinical management of risk. PMID- 10789280 TI - Axis I comorbidity in bipolar disorder with psychotic features. AB - BACKGROUND: Axis I comorbidities are prevalent among patients with severe bipolar disorder but the clinical and psychopathological implications are not clear. AIMS: To investigate characteristics of four groups of patients categorised as follows: substance abuse only (group 1), substance abuse associated with other Axis I disorders (group 2), non-substance-abuse Axis I comorbidity (group 3), no psychiatric comorbidity (group 4). METHOD: Consecutive patients with bipolar disorder with psychotic features (n = 125) were assessed using the Structured Clinical Interview for DSM-III-R--patient version, and several psychopathological scales. RESULTS: By comparison with group 4, group 1 had a higher risk of having mood-incongruent delusions, group 2 had an earlier age at onset of mood disorder, a more frequent onset with a mixed state and a higher risk of suicide, and group 3 had more severe anxiety and a better awareness of illness. CONCLUSIONS: Substance abuse, non-substance-abuse Axis I comorbidity and their reciprocal association are associated with different characteristics of bipolar disorder. PMID- 10789282 TI - Evaluation of ICD-10 PHC mental health guidelines in detecting and managing depression within primary care. AB - BACKGROUND: The prevalence of mental disorders in the community is high, yet many remain unrecognised, misdiagnosed or poorly managed within primary care. Hence, guidelines for diagnosing and managing mental disorders in primary care, ICD-10 PHC, have been developed. AIMS: To introduce the guidelines into primary care and to assess whether they improve recognition, accuracy of diagnosis and treatment standards. METHOD: GPs recorded information on all patients presenting with mental disorders before and after guidelines were introduced. A 10% sample of patients underwent interviews to establish a formal diagnosis. Recognition of mental disorders was assessed by screening of patients attending their GP. RESULTS: The guidelines had no impact on the overall detection of mental disorders, the accuracy of diagnosis or the prescription of antidepressants. There was a significant increase in the number of patients diagnosed with depression or unexplained somatic symptoms. The GPs also made increased use of psychological interventions. CONCLUSIONS: The success of the guidelines in bringing about change is by no means certain. Some areas appeared more susceptible to change than others. PMID- 10789281 TI - Seasonal affective disorder among primary care attenders and a community sample in Aberdeen. AB - BACKGROUND: There are no large published studies of the prevalence of seasonal affective disorder (SAD) among UK populations. AIM: To determine the prevalence of SAD among patients attending a general practitioner (GP). METHOD: Patients aged 16-64 consulting their GPs in Aberdeen during January were screened with the Seasonal Pattern Assessment Questionnaire (SPAQ). SPAQs were also mailed to 600 matched patients, who had not consulted their GP during January. Surgery attenders who fulfilled SPAQ criteria for SAD were invited for interview to determine whether they met criteria for SAD in DSM-IV and the Structured Interview Guide for the Hamilton Rating Scale for Depression--Seasonal Affective Disorder Version (SIGH-SAD). RESULTS: Of 6161 surgery attenders, 4557 (74%) completed a SPAQ; 442 (9.7%) were SPAQ cases of SAD. Rate of caseness on the SPAQ did not differ between surgery attenders and non-attenders. Of 223 interviewed SPAQ cases of SAD, 91 (41%) also fulfilled DSM-IV and SIGH-SAD criteria. CONCLUSIONS: There is a high prevalence of SAD among patients attending their GPs in January in Aberdeen; this is likely to reflect a similar rate in the community. PMID- 10789283 TI - 'Obsessions' in children with autism or Asperger syndrome. Content analysis in terms of core domains of cognition. AB - BACKGROUND: We report a survey of the content of obsessions in children with autism spectrum conditions. We use the term 'obsessions' narrowly, to indicate strong, repetitive interests. We predicted that obsessions would not cluster randomly, but rather would occur significantly more often in the domain of 'folk physics' (an interest in how things work), and significantly less often in the domain of 'folk psychology' (an interest in how people work). These predictions were tested relative to a control group of 33 children with Tourette syndrome. AIMS: To examine the content of autistic obsessions, and to test the theory that these reflect an evolved cognitive style of good folk physics alongside impaired folk psychology. METHOD: Ninety-two parents returned a questionnaire designed to determine the subject of their child's obsessional interests. The results were analysed in terms of core domains of cognition. RESULTS: Both predictions were confirmed. CONCLUSIONS: These results suggest that impaired folk psychology and superior folk physics are part of the cognitive phenotype of autism. A content free theory of obsessions is inadequate. PMID- 10789284 TI - Sertraline-induced anorgasmia reversed by nefazodone. PMID- 10789285 TI - Acute manic symptomatology during repetitive transcranial magnetic stimulation in a patient with bipolar depression. PMID- 10789286 TI - Cortical and spinal excitability. PMID- 10789287 TI - Ethnicity and first year of contact with psychiatric services. PMID- 10789288 TI - Psychiatry and homosexuality. PMID- 10789289 TI - Psychiatry and homosexuality. PMID- 10789290 TI - Old age forensic psychiatry. PMID- 10789291 TI - The confidential inquiry comes of age. PMID- 10789292 TI - Variations in psychiatric practice. Neither unacceptable nor unavoidable, only under-researched. PMID- 10789293 TI - Providing specialised services for anorexia nervosa. PMID- 10789294 TI - Guiding principles of drug demand reduction: an international response. AB - BACKGROUND: The 20th century has seen the globalisation and homogenisation of substance misuse problems, blurring traditional boundaries between producer and consumer countries and forcing the international community to consider new responses to substance misuse. AIMS: To highlight the importance of the principles of illicit drug demand reduction and related activities in tackling global drug problems and to discuss the commitment made by United Nations (UN) Member States at the highest political level towards meeting the objectives set in their Declaration on the Guiding Principles. METHOD: A review of international responses to substance misuse. RESULTS: Traditionally, the focus has been on reducing the supply of controlled drugs, while maintaining adequate supplies of narcotic and psychotropic drugs for clinical and scientific purposes. However, it has become apparent that supply reduction in isolation is insufficient and demand reduction is now receiving greater emphasis--culminating in the UN General Assembly adopting the Declaration on the Guiding Principles of Demand Reduction in 1998. CONCLUSIONS: This declaration offers a genuinely holistic approach to the complexities of substance misuse. To be successful, it requires the commitment of governments at the highest level and the willing participation of small communities. PMID- 10789295 TI - Outcome of admission to a medium secure psychiatric unit. I. Short- and long-term outcome. AB - BACKGROUND: This is the first long-term follow-up of patients discharged from a medium secure unit. AIMS: To describe the short- and long-term outcomes of admission for all patients discharged during a 14-year period. METHOD: A longitudinal cohort study of all 234 patients discharged from the Denis Hill Unit, Bethlem Royal Hospital, between 1980 and 1994, followed for an average 6.6 years. RESULTS: Although 48% of admissions were from prison, only 8% returned there, with most being transferred to another psychiatric bed. One-fifth of patients spent none of the follow-up time in the community; 75% of patients had at least one readmission; only 24% were convicted of further offences. CONCLUSIONS: Re-offending rates are comparable with those for patients discharged from high-security hospitals, and much lower than those for released prisoners. The high readmission rates indicate the need for a range of services to maintain former patients in the community. PMID- 10789296 TI - Outcome of admission to a medium secure psychiatric unit. 2. Role of ethnic origin. AB - BACKGROUND: A follow-up of patients discharged from medium secure psychiatric units is used to compare outcome in patients of different ethnic origin. AIMS: To test the hypothesis that there are systematic differences in clinical outcome between ethnic groups. METHOD: A descriptive, longitudinal cohort study of discharges from a medium secure unit is used to compare the 125 patients of White/European ethnic origin and the 104 patients of Black/African-Caribbean origin. RESULTS: Patients of African-Caribbean origin were admitted at three times the rate of White patients, had a higher prevalence of psychosis and a lower prevalence of personality disorder. There was no difference in outcome as measured by location at follow-up, readmission or re-offending. CONCLUSIONS: The higher rate of admission of African-Caribbean patients is consistent with a higher level of demand. There is a need for studies of the pathways by which patients from ethnic minorities reach medium-security accommodation, with a view to early intervention. PMID- 10789297 TI - Case register study of suicide risk in mental disorders. AB - BACKGROUND: There have been few large-scale studies of long-term suicide risk in mental disorders in the UK. AIMS: To estimate the long-term risk of suicide in psychiatric patients. METHOD: A sample of 7921 individuals was identified from the Salford Psychiatric Case Register. Mortality by suicide or undetermined external cause during a follow-up period of up to 18 years was determined using the NHS Central Register; suicide risks were estimated as rate ratios. RESULTS: Suicide risk was increased more than ten-fold in both genders: the rate ratio for males was 11.4; for females it was 13.7. The risk was highest in young patients, but high risk continued into late life. The diagnoses with the highest risk were schizophrenia, affective disorders, personality disorder and (in males) substance dependence. Risk was also associated with recent initial contact and number of admissions but not comorbidity. CONCLUSIONS: The suicide risks estimated in this study are generally higher than those previously reported, notably in schizophrenia and personality disorder, and in previous in-patients. Patients with these high-risk diagnoses, an onset of illness within the previous 1-3 years, or more than one previous admission should be regarded as priority groups for suicide prevention by mental health services. PMID- 10789298 TI - Evaluating mental health policy in England. Care Programme Approach and supervision registers. AB - BACKGROUND: The Care Programme Approach (CPA) and supervision register policies in England are intended to prioritize patients to receive specialist mental health care. AIMS: To describe and evaluate the practical application of the policies. METHOD: A questionnaire survey of key informants in mental health provider trusts and an analysis of aggregated data collected by health authorities using the Mental Illness Needs Index as a measure of population need. RESULTS: On average, 1175 per 100,000 total population are subject to the CPA (95% CI = 1055-1309) and 8.6 per 100,000 (95% CI = 7.5-9.9) are on supervision registers. Wide local variations in the number of people subject to the CPA and supervision registers are not explained by variations in population need. CONCLUSIONS: Prioritization to receive specialist mental health services is carried out inconsistently, and inequitable use of resources may result. PMID- 10789299 TI - Effectiveness of cognitive therapy for delusions in routine clinical practice. AB - BACKGROUND: Previous studies have suggested that cognitive therapy is effective in modifying delusions. AIMS: To assess the effectiveness of cognitive therapy on patients seen in routine clinical work. METHOD: Eighteen patients with chronic delusions were treated using cognitive therapy, after the method of Chadwick and Lowe. A single-case multiple-baseline experimental design was used, including a control treatment. Each subject was used as their own control. RESULTS: Six patients reduced conviction in their delusions during cognitive therapy and not during the control treatment. Seven patients' conviction ratings did not change. Five patients showed a variable response. Degree of conviction did not fall to zero in any patient. All patients reported that the therapy had been helpful; six spontaneously mentioned changes in psychotic thinking. CONCLUSIONS: One-third of patients with chronic delusions whom we treated responded to delusion modification with a reduction in degree of belief. Change within therapy sessions predicted outcome, as did variation in the conviction during baseline. Cognitive therapy with delusions should aim at reducing distress as well as conviction. PMID- 10789300 TI - Lithium-associated clinical hypothyroidism. Prevalence and risk factors. AB - BACKGROUND: Rates of, and risk factors for, lithium-associated clinical hypothyroidism are uncertain. AIMS: To determine prevalence of and risk factors for clinical hypothyroidism in patients treated with lithium carbonate. METHOD: Retrospective case-note review of 718 patients who had undergone serum lithium estimation during a 15-month period. Patients on thyroxine had a more detailed review. RESULTS: The prevalence of clinical hypothyroidism during lithium treatment was 10.4%. The main risk factor was female gender (women 14% v. men 4.5%). Women were at highest risk during the first two years of lithium treatment, and women starting lithium aged 40-59 years had the greatest prevalence (> 20%). No equivalent risk factors emerged in men, although, like women, their prevalence of hypothyroidism was substantially higher than community rates. CONCLUSIONS: The high rates of clinical hypothyroidism identified may call for a review of the drug information given to women, particularly to those starting lithium in middle age. Consideration should be given to screening for thyroid antibodies before treatment in high-risk cases. Monitoring of thyroid function should take into account gender, age and stage of lithium treatment. PMID- 10789301 TI - Community-based case-control study of depression in older people. Cases and sub cases from the MRC-ALPHA Study. AB - BACKGROUND: Risk factors of depression in later life, particularly for sub-cases and for psychotic and neurotic types of depression, are unclear. AIMS: To identify such risk factors. METHOD: Over 5200 older people (> or = 65 years), randomly selected from Liverpool, were interviewed using the Geriatric Mental State (GMS) and the Minimum Data Set (MDS). The computer-assisted diagnosis AGECAT identified 483 cases and 575 sub-cases of depression and 2451 with no mental problems. Logistic regression was employed to examine factors relevant to caseness. RESULTS: In multiple logistical regression, odds ratios (ORs) were significantly high for being female (2.04, 95% CI 1.56-2.69), widowed (2.00, 1.18 3.39), having alcohol problems (4.37, 1.40-2.94), physical disablement (2.03, 1.40-2.94), physical illness (1.98, 1.25-3.15), taking medications to calm down (10.04, 6.41-15.71), and dissatisfaction with life (moderate 4.54, 3.50-5.90; more severe 29.00, 16.00-52.59). Good social networks reduced the ORs. If sub cases were included as controls, the statistical significance was reduced. CONCLUSIONS: Age was not associated with depression in later life whereas gender, physical disablement and dissatisfaction with life were. The sub-cases shared many risk factors with cases, suggesting that prevention may need to be attempted at an early stage. PMID- 10789302 TI - Bereavement and grief in adults with learning disabilities. A follow-up study. AB - BACKGROUND: This paper reports on the follow-up of a cohort of parentally bereaved adults with learning disabilities. AIMS: To investigate whether significant psychopathology, present up to 2.1 years after the death, had resolved five years later. METHOD: Of an original sample of 50 adults with learning disabilities, 41 were reassessed. The Aberrant Behaviour Checklist and the Psychopathology Instrument for Mentally Retarded Adults were re-administered to carers. RESULTS: At follow-up, there was a small increase in the measures of aberrant behaviour. Measures of psychopathology showed improvement, and in particular there was a reduction in anxiety. CONCLUSIONS: The response to bereavement by adults with learning disabilities is similar in type, though not in expression, to that of the general population. Learning disability is a significant predictor of mental health problems following bereavement. Participants adapted more easily when basic emotional needs had been constructively met by carers. PMID- 10789303 TI - Genetic and environmental risk factors in the aetiology of illicit drug initiation and subsequent misuse in women. AB - BACKGROUND: Subsequent to initial exposure to the use of a psychoactive substance, psychoactive substance use disorder (PSUD) may or may not develop. AIMS: To investigate the relationship between the risk factors for initiation and the subsequent misuse of psychoactive substances. METHOD: The lifetime history of illicit substance use and misuse was obtained by telephone interview with 1934 members of female-female twin pairs. We apply a novel model, which estimates the role of genetic and environmental risk factors that influence initiation and those specific to misuse, to three classes of illicit psychoactive substances. RESULTS: The individual-specific environment and family environment influenced the probability of initiation, but only individual-specific environment had an impact on the probability of subsequent misuse. Genetic factors which influence the risk of initiation and of misuse were identified. CONCLUSIONS: Aetiological factors that influence drug initiation and subsequent misuse are correlated but not identical. Family environment is an important determinant of risk for drug experimentation. Two classes of genetic risk factors act on the liability to PSUD: those that influence the probability of initiation and those that influence the risk of misuse. PMID- 10789304 TI - Altered cerebral perfusion measured by SPECT in relatives of patients with schizophrenia. Correlations with memory and P300. AB - BACKGROUND: Genetic studies in schizophrenia are hampered by the complex heterogeneous clinical phenotype. Biological variables identified as trait markers of risk could clarify the mode of inheritance, define clinical subgroups and provide clues about aetiology. AIMS: To use single photon emission computed tomography (SPECT) to compare brain perfusion maps in patients with schizophrenia (n = 19), their asymptomatic 'high-risk' relatives (n = 36) and control subjects (n = 34) and to examine the relationships between imaging, memory and P300 event related potential. METHOD: SPECT, memory tests and P300 recording were carried out. RESULTS: In the patients with schizophrenia and their relatives, perfusion was reduced in left inferior prefrontal and anterior cingulate cortex and increased bilaterally in a subcortical region. Perfusion significantly correlated with verbal memory and P300 amplitude in left inferior prefrontal cortex and with P300 latency in anterior cingulate cortex. CONCLUSIONS: Medication- and symptom free relatives had altered regional perfusion intermediate between subjects with schizophrenia and controls. Impaired perfusion, verbal memory and P300 appear to be related traits associated with an increased risk of illness. PMID- 10789306 TI - Transcallosal inhibition and motor conduction studies in patients with schizophrenia using transcranial magnetic stimulation. AB - BACKGROUND: Transcranial magnetic stimulation of the motor cortex may not only elicit excitatory responses in hand muscles contralateral to the stimulated hemisphere, but may also suppress tonic voluntary electromyogram activity in muscles ipsilateral to the stimulation. This inhibition is mediated between the motor cortices via the corpus callosum. AIMS: To investigate motor excitability and interhemispheric (transcallosal) connections in patients with schizophrenia. METHOD: Transcallosal inhibition and motor conduction parameters were investigated in ten patients with schizophrenia and in ten age- and gender matched healthy subjects. RESULTS: Transcallosal conduction time (TCT) and duration of the inhibition were significantly longer in patients with schizophrenia (mean (s.d.)): TCT, 12.4 (2.9) ms in normal subjects and 15.3 (2.6) ms in patients (P = 0.03); mean duration, 34.1 (4.9) ms in normal subjects and 51.9 (16.8) ms in patients (P = 0.01). CONCLUSIONS: Magnetic motor conduction parameters are unaltered in schizophrenia, but transcallosal inhibition is significantly delayed and prolonged. This may indicate abnormal function of the corpus callosum in these patients. PMID- 10789305 TI - No evidence for left superior temporal dysfunction in asymptomatic schizophrenia and bipolar disorder. PET study of verbal fluency. AB - BACKGROUND: Positron emission tomography (PET) studies have revealed functional left superior temporal gyrus (STG) abnormalities in symptomatic schizophrenia during word generation. AIMS: To discover if this dysfunction is present in asymptomatic schizophrenia. To determine whether, without concurrent symptomatology, schizophrenia and bipolar affective disorder (BPD) are distinguishable by differing regional cerebral blood flow (rCBF) patterns during word generation. METHOD: A PET verbal fluency protocol was applied to six patients with BPD in remission and six patients with asymptomatic schizophrenia. Analysis included 10 control subjects from a contemporaneous study. RESULTS: All groups showed relative reduction of rCBF in both superior temporal cortices. There were no quantitative differences in any group comparison. All groups exhibited negative covariation between rCBF in left prefrontal and right (but not left) temporal regions. CONCLUSIONS: Abnormal patterns of left STG function cannot be regarded as a trait marker for schizophrenia. Functional abnormalities may reflect aspects of mental state. PMID- 10789308 TI - Volume of parahippocampal gyrus and hippocampus in schizophrenia. PMID- 10789307 TI - Sensorimotor and cognitive development of infants of mothers with schizophrenia. AB - BACKGROUND: The parenting environment provided by mothers with schizophrenia is likely to contribute to the cognitive impairment observed in their offspring. AIMS: To assess the relative contribution of maternal schizophrenia, obstetric factors and mothers' lifestyle to the cognitive development of infants in their first year. METHOD: The Bayley Scales of Infant Development were administered to 19 infants of mothers with schizophrenia, 34 with non-psychotic depression in the mother, 29 with affective psychosis in the mother and 24 normal controls when the babies were two and seven months of age. RESULTS: At two months, infants of mothers with schizophrenia and of those with affective psychosis had lower mental development index (MDI) scores than controls. At seven months, infants of mothers with schizophrenia had lower MDI scores than all other groups. When infant birth weight and mothers' social class were taken into account there was no effect of maternal schizophrenia on infants' MDI scores. CONCLUSIONS: Infants of mothers with schizophrenia are likely to have impaired cognitive development. This may be due in part to environmental factors such as the mother's lifestyle. PMID- 10789309 TI - Post-abortion mania. PMID- 10789310 TI - Mirtazapine withdrawal causing hypomania. PMID- 10789311 TI - Drug therapy in treatment-resistant depression. PMID- 10789312 TI - Pharmacokinetics of clozapine. PMID- 10789313 TI - Olanzapine and tardive dyskinesia. PMID- 10789314 TI - Atypical antipsychotics and neuroleptic malignant syndrome. PMID- 10789315 TI - Prescribing donepezil in clinical practice. PMID- 10789316 TI - More trouble with ethics committees. PMID- 10789317 TI - Is psychiatry more mindful or brainier than it was a decade ago? PMID- 10789318 TI - The next 25 years. PMID- 10789319 TI - Three decades after Penrose. PMID- 10789320 TI - Genetics advances and learning disability. AB - BACKGROUND: Medicine is rapidly becoming molecular medicine, and little escapes the grasp of modern genetics. Most disorders associated with learning disability have at least a genetic component influencing their expression; in many disorders, disturbances of genetic mechanisms play a pivotal role. AIMS: Dynamic mutations, imprinting mechanisms and gene-dosage effects are explained with reference to genetic disorders that lead to learning disability. METHOD: A review of recent important studies in the genetics of learning disability. RESULTS: A host of new genetic connections to conditions associated with learning disability have been made. CONCLUSIONS: A basic understanding of these genetic connections is important for all learning disability psychiatrists if they are to follow the rapid changes--already beginning to influence our practice--that hold immense promise for the future. PMID- 10789321 TI - Autism--an evolving concept. AB - BACKGROUND: The rapid increase in research endeavour has not kept pace with the advent of well-publicized theories and treatments for autism. AIMS: To explore some of the newer developments in biological research into autism. METHOD: A review of recent publications and presentations. RESULTS: The concept is shifting from the narrow perception of aloof autism, described by Kanner, to a wider one that includes a spectrum extending to a broader, subclinical phenotype. The genetic basis has been established; now we need to discover the location and interaction of the relevant sites. There is considerable interest in the bowel as a pathogenetic agent, particularly in the effects of exogenous opioids and multiple viral infection (the latter posing a public health problem). Also of concern is the role of (potentially treatable) epilepsy, analogous to the Laudau Kleffner syndrome. CONCLUSIONS: In the absence of a cure, the implementation of ideas will continue to outstrip factual evidence. Clinicians are challenged by the availability of information (and misinformation), particularly on the internet. PMID- 10789322 TI - Ageing and learning disability. AB - BACKGROUND: Ageing is a continuation of the developmental process and is influenced by genetic and other biological factors as well as personal and social circumstances. AIMS: To identify some key biological, psychological and social issues relevant to how ageing might particularly effect people with learning disabilities. METHOD: This selected review considers the extent to which there are similarities and differences relative to people without learning disabilities. RESULTS: There is a convergence, in later life, between people with a learning disability and those without, owing to the reduced life expectancy of people with more severe disabilities. People with Down's syndrome have particular risks of age-related problems relatively early in life. CONCLUSIONS: The improved life expectancy of people with learning disabilities is well established. There is a lack of a concerted response to ensure that the best possible health and social care is provided for people with learning disabilities in later life. PMID- 10789323 TI - Psychotherapy, learning disabilities and trauma: new perspectives. AB - BACKGROUND: Psychological therapies are rarely used in people with learning disabilities. Learning disability is often given as an exclusion criterion. There is insufficient published research to conduct a systematic review. Few outcome studies of psychoanalytic or cognitive psychotherapy have been reported. AIMS: To describe recent advances in understanding and practice within the learning disability field which have not received wider recognition within mainstream psychotherapy and psychiatry. METHOD: The availability of different psychotherapeutic approaches is discussed. We explore developmental issues including the contribution of attachment theory to our understanding, and the effects of trauma on the lives of people with learning disabilities. RESULTS: Theoretical and clinical perspectives suggest that many therapeutic opportunities exist. CONCLUSIONS: Practitioners are encouraged to extend their therapeutic repertoire, and to report measurable outcomes. PMID- 10789324 TI - Health care provision for people with a learning disability. Record-linkage study of epidemiology and factors contributing to hospital care uptake. AB - BACKGROUND: We know little about how people with a learning disability access secondary health care. AIMS: To describe the epidemiology of learning disability, the influence of deprivation on prevalence and the pattern of secondary care uptake, including the effect of institutionalisation. METHOD: A record-linkage study of secondary care contacts of 434,000 people between 1991 and 1997. A population with learning disability was identified; their secondary care contact was calculated and compared with the general population's. RESULTS: The distribution of people with a learning disability (n = 1595) correlated significantly with deprivation. The presence of a learning disability hospital significantly affected care uptake. Place of residence also affected acute admission to the learning disability hospital. Former institution residents generated 212 admissions per 1000 patients; community patients generated 18 per 1000. The admission rate with any psychiatric diagnosis to any setting was 26.3 per 1000 people with a learning disability; 16.5% of such patients had a dual diagnosis. CONCLUSIONS: Health provision for people with a learning disability is affected by institutional provision. PMID- 10789325 TI - Reducing antipsychotic medication in people with a learning disability. AB - BACKGROUND: The use of antipsychotic drugs in people with learning disabilities is currently receiving intensified scrutiny and attempts are being made to reduce it. AIMS: A randomised controlled trial was designed to investigate factors influencing antipsychotic drug reduction among people with learning disabilities prescribed such medication for behavioural problems. METHOD: Thirty-six participants randomly allocated to the experimental group underwent four, monthly 25% drug reduction stages. There were no planned drug changes for the control group (n = 20). RESULTS: Twelve participants (33%) completed full withdrawal; a further seven (19%) achieved and maintained at least a 50% reduction. Drug reduction was associated with increased dyskinesia and higher activity engagement but not increased maladaptive behaviour. Some setting characteristics were associated with drug reinstatement. CONCLUSIONS: A substantial proportion of people with learning disability prescribed antipsychotic medications for behavioural purposes rather than for treating psychotic illness can have their drugs reduced or withdrawn. PMID- 10789326 TI - Open peer review: a randomised controlled trial. AB - BACKGROUND: Most scientific journals practise anonymous peer review. There is no evidence, however, that this is any better than an open system. AIMS: To evaluate the feasibility of an open peer review system. METHOD: Reviewers for the British Journal of Psychiatry were asked whether they would agree to have their name revealed to the authors whose papers they review; 408 manuscripts assigned to reviewers who agreed were randomised to signed or unsigned groups. We measured review quality, tone, recommendation for publication and time taken to complete each review. RESULTS: A total of 245 reviewers (76%) agreed to sign. Signed reviews were of higher quality, were more courteous and took longer to complete than unsigned reviews. Reviewers who signed were more likely to recommend publication. CONCLUSIONS: This study supports the feasibility of an open peer review system and identifies such a system's potential drawbacks. PMID- 10789327 TI - Functional anatomy of verbal fluency in people with schizophrenia and those at genetic risk. Focal dysfunction and distributed disconnectivity reappraised. AB - BACKGROUND: PET studies of verbal fluency in schizophrenia report a failure of 'deactivation' of left superior temporal gyrus (STG) in the presence of activation of left dorsolateral prefrontal cortex (DLPFC), which deficit has been attributed to underlying 'functional disconnectivity'. AIM: To test whether these findings provide trait-markers for schizophrenia. METHOD: We used H2(15)O PET to examine verbal fluency in 10 obligate carriers of the predisposition to schizophrenia, 10 stable patients and 10 normal controls. RESULTS: We found no evidence of a failure of left STG deactivation in carriers or patients. Instead, patients failed to deactivate the precuneus relative to other groups. We found no differences in functional connectivity between left DLPFC and left STG but patients exhibited significant disconnectivity between left DLPFC and anterior cingulate cortex. CONCLUSIONS: Failure of left STG 'deactivation' and left fronto temporal disconnectivity are not consistent findings in schizophrenia; neither are they trait-markers for genetic risk. Prefrontal functional disconnectivity here may characterise the schizophrenic phenotype. PMID- 10789328 TI - Invited commentary on: functional anatomy of verbal fluency in people with schizophrenia and those at genetic risk. The genetics of asymmetry and psychosis. PMID- 10789330 TI - Season of birth in schizophrenia: no latitude at the equator. AB - BACKGROUND: If the established winter excess in births of people who subsequently develop schizophrenia is an effect of 'seasonality', this would be testable by examining the pattern of births in an equatorial region with no formal seasons. AIMS: To investigate whether there is any variation in month of birth among patients from equatorial Singapore with a diagnosis of schizophrenia. METHOD: All 9655 patients discharged from Singapore's national psychiatric hospital with a diagnosis of schizophrenia were included (year of birth range 1930-1984). We analysed aggregated data, as well as the data of subsamples grouped according to birth-year periods, in order to examine secular trends. One patient subsample (those born 1960-84) allowed exact matching against the general population data set and close testing of any seasonal influence. RESULTS: Monthly variation in births was evident for both patients and controls; the patterns were very similar, apart from the patient sample showing a trough in March-April. CONCLUSIONS: In an equatorial region, where 'seasons' are absent, no seasonal excess in births of those later developing schizophrenia was evident. PMID- 10789329 TI - Changes in suicide in England and Wales, 1960-1997. AB - BACKGROUND: Methods of suicide and suicide rates in England and Wales have fluctuated considerably since the 1960s. AIMS: To review the changes that have occurred in suicide rates in England and Wales between 1960 and 1997. METHOD: Suicide rates, derived from total annual suicides and the estimated annual resident population, were obtained from the Office for National Statistics. RESULTS: Suicide rates decreased in both genders between the early 1960s and the mid-1970s. The rate for males then increased between 1975 and 1990, while the rate for females continued to fall. Between 1990 and 1997, the rate decreased for males and females in all age groups, particularly for those using motor vehicle exhaust gas; the latter finding is associated with increasing use of catalytic converters. CONCLUSIONS: Following the increase in suicide among males until 1990 there was a decrease for both genders between 1990 and 1997, consistent with the 'Health of the Nation' target. PMID- 10789331 TI - Impaired regulation of brain serotonin function during dieting in women recovered from depression. AB - BACKGROUND: Amino acid mixtures that lower brain availability of the serotonin (5 HT) precursor tryptophan produce acute depressive relapse in women with a history of major depression. Dieting also lowers brain tryptophan availability, but its effects on brain 5-HT function in recovered depressed women have not been studied. AIM: To test the hypothesis that women with a history of major depression would show impaired regulation of brain 5-HT function during a period of dieting-induced tryptophan depletion. METHOD: Women with and without a history of major depression were placed on a daily 1000 kcal (approximately 4200 kj) diet for three weeks. Before the diet and in the final week we measured fasting plasma tryptophan levels and the prolactin response to an intravenous tryptophan challenge. RESULTS: Dieting lowered plasma tryptophan levels equivalently in women with and without a history of depression. In women without a history of depression, dieting also increased the prolactin response to tryptophan. This increase did not occur in women with a history of depression. CONCLUSIONS: Women with a history of depression showed impaired regulation of brain 5-HT function in response to dieting. PMID- 10789332 TI - Mortality and causes of death in a total national sample of patients with affective disorders admitted for the first time between 1973 and 1993. AB - BACKGROUND: The high mortality from suicide in patients admitted to hospital with an affective disorder is well documented, although specific causes of mortality and changes in mortality are less well studied. AIMS: To describe the pattern of mortality in patients with affective disorder and to study changes in suicide risk during the study period. METHOD: All patients (n = 54,103) admitted for the first time to a psychiatric hospital in Denmark during the period 1973-1993 because of affective disorder were included in this study. The mortality rate was compared with that of the general population. RESULTS: Mortality from natural and unnatural causes was elevated in all subgroups of affective disorder. The risk of suicide among patients ill for one year or less after first admission increased during the period 1973-1993. CONCLUSIONS: More attention should be paid to the risk of suicide and to physical illness in patients with affective disorders. PMID- 10789333 TI - Tokophobia: an unreasoning dread of childbirth. A series of 26 cases. AB - BACKGROUND: Some women dread and avoid childbirth despite desperately wanting a baby. This is called tokophobia. AIMS: To classify tokophobia for the first time in the medical literature. METHOD: Twenty-six women noted to have an unreasoning dread of childbirth were interviewed by the same psychiatrist, who was not the treating doctor. A qualitative analysis of these psychiatric interviews was performed. RESULTS: Phobic avoidance of pregnancy may date from adolescence (primary tokophobia), be secondary to a traumatic delivery (secondary tokophobia) or be a symptom of prenatal depression (tokophobia as a symptom of depression). Pregnant women with tokophobia who were refused their choice of delivery method suffered higher rates of psychological morbidity than those who achieved their desired delivery method. CONCLUSIONS: Tokophobia is a specific and harrowing condition that needs acknowledging. Close liaison between the obstetrician and the psychiatrist in order to assess the balance between surgical and psychiatric morbidity is imperative with tokophobia. PMID- 10789335 TI - Crisis telephone consultation for deliberate self-harm patients. PMID- 10789334 TI - Jerusalem syndrome. AB - BACKGROUND: Jerusalem's psychiatrists expect to encounter, as the millennium approaches, an ever-increasing number of tourists who, upon arriving in Jerusalem, may suffer psychotic decompensation. AIMS: To describe the Jerusalem syndrome as a unique acute psychotic state. METHOD: This analysis is based on accumulated clinical experience and phenomenological data consisting of cultural and religious perspectives. RESULTS: Three main categories of the syndrome are identified and described, with special focus on the category pertaining to spontaneous manifestations, unconfounded by previous psychotic history or psychopathology. CONCLUSIONS: The discrete form of the Jerusalem syndrome is related to religious excitement induced by proximity to the holy places of Jerusalem, and is indicated by seven characteristic sequential stages. PMID- 10789336 TI - Suicide attempts v. deliberate self-harm: a response. PMID- 10789337 TI - Post-abortion mania. PMID- 10789338 TI - Cognitive effects of antipsychotics in schizophrenia and relationship to quality of life. PMID- 10789339 TI - Antidepressant response reversed by interferon. PMID- 10789340 TI - Post-stroke depression. PMID- 10789341 TI - Correlation between cerebral perfusion and depressive symptom scores from morning to evening. PMID- 10789342 TI - Lithium-induced hypothyroidism. PMID- 10789343 TI - Focus on psychiatry in Australia. PMID- 10789344 TI - PRiSM Psychosis Study. Design limitations, questionable conclusions. PMID- 10789345 TI - PRiSM Psychosis Study. Thro'a glass darkly: a distorted appraisal of community care. PMID- 10789346 TI - Community mental health teams: evidence or belief? PMID- 10789347 TI - The EPSILON study of schizophrenia in five European countries. Design and methodology for standardising outcome measures and comparing patterns of care and service costs. AB - BACKGROUND: There is a lack of cross-national research on care for people with schizophrenia. AIMS: To produce standardised European versions of five instruments in key areas of mental health service research in five languages, and to compare data from five European countries regarding patients with schizophrenia and mental health care provision and costs. METHOD: Five centres, in Amsterdam, Copenhagen, London, Santander and Verona, participated. Instruments assessing needs, service use, informal career involvement, quality of life, and service satisfaction were subjected to a conversion procedure including translation, back-translation, focus group discussion and reliability assessment. Patients of local mental health services with a Schedules for Clinical Assessment in Neuropsychiatry diagnosis of schizophrenia were interviewed. RESULTS: Service provision varied between sites; 404 patients were studied. Instrument reliability was found to be good. CONCLUSIONS: The instruments developed were reliable across the range of countries, and will facilitate future comparative health service research. PMID- 10789348 TI - Annual variation in birth rate of people who subsequently develop schizophrenia. AB - BACKGROUND: Previous attempts to decide whether there is significant year-to-year variation in the birth rate of people who subsequently develop schizophrenia have given conflicting results, probably because of differences in the statistical methods employed. AIMS: To determine whether there is significant year-to-year variation in the birth rate. METHOD: Variation in the birth rate for the period 1921-1960 was studied in three separate national data sets--English, Danish and Scottish--using cubic splines to smooth the distribution curve before calculating residuals from a Poisson distribution. RESULTS: Over-dispersion was found in all three data sets, particularly in the Danish and Scottish data. However, the correlation between the sets of standardised residuals derived from the three data sets was only statistically significant for Denmark v. England. CONCLUSIONS: There was statistically significant year-to-year variation in the birth rate of people who subsequently developed schizophrenia in three countries in north-west Europe in the years 1921-1960. This is potentially a clue to the nature of the environmental determinants of schizophrenia, but better data will be needed before useful explanatory hypotheses can be generated and tested. PMID- 10789349 TI - Patients with personality disorder admitted to secure forensic psychiatry services. AB - BACKGROUND: Treatment of patients with personality disorder remains controversial and severe mental illness is prioritized in secure forensic psychiatry services. AIMS: To compare patients with personality disorder and mental illness according to demography, referral, criminality, previous institutionalisation and diagnostic comorbidity. METHOD: A record survey of 511 patients with personality disorder and 2575 with mental illness admitted to secure forensic psychiatry services between 1 January 1988 and 31 December 1994 from half of England and Wales. RESULTS: Personality disorder admissions declined over time; more were female, White, younger and extensively criminal (specifically, sexual and arson offences). Personality disorder was highly comorbid; antisocial, borderline, paranoid and dependent personality disorder were most prevalent. CONCLUSIONS: Patients with personality disorder were highly selected and previously known to psychiatric services. Referrer, diagnostic comorbidity and behavioural presentation determined their pathways into care. Future research must determine whether their continuing admission represents effective use of scarce resources and whether new services are required. PMID- 10789350 TI - Diagnostic stability of first-episode psychosis. Comparison of ICD-10 and DSM-III R systems. AB - BACKGROUND: The temporal stability of a diagnosis is one measure of its predictive validity. AIMS: To measure diagnostic stability in first-episode psychosis using ICD-10 and DSM-III-R. METHOD: Between 1992 and 1994 we ascertained a cohort of persons with first-episode psychosis (n = 168), assigning to each a consensus diagnosis. At three-year follow-up, longitudinal consensus diagnoses, blind to onset diagnoses, were made. Stability was measured by the positive predictive values (PPVs) of onset diagnoses. For onset schizophrenia, we also calculated sensitivity, specificity and concordance (kappa). RESULTS: First episode ICD-10 and DSM-III-R schizophrenia had a PPV of over 80% at three years. Over one-third of cases with ICD-10 F20 schizophrenia at three years had non schizophrenia diagnoses at onset. Manic psychoses showed the highest PPV (91%). For onset schizophrenia, both systems had high specificity (ICD-10: 89; DSM-III R: 93%), but low sensitivity (ICD-10: 64%; DSM-III-R: 51%) and moderate concordance (ICD-10: 0.54; DSM-III-R: 0.46). CONCLUSIONS: Bipolar disorders and schizophrenia showed the highest stability. DSM-III-R schizophrenia did not have greater stability than ICD-10 schizophrenia. PMID- 10789351 TI - More severe mental illness is more concentrated in deprived areas. AB - BACKGROUND: The greater frequency of mental illness in deprived and inner-city populations is well recognised; allocation of funds in the UK health service makes some allowance for this. However, it is not clear whether the differences are similar for all levels of mental health care need. AIMS: To study the range in prevalence of mental health problems and care at primary care, general secondary care and forensic care levels. METHOD: We used mainly descriptive statistics to study evidence available from existing sources--some based on indicators of likely need, some on observed prevalence of treatment. RESULTS: Among English health authority areas, the most morbid have about twice the prevalence of primary care level mental illness of the least morbid. For secondary care the ratio is between 2.5 and 4 to 1, while for services for mentally disordered offenders it is in excess of 20:1. CONCLUSIONS: Where needs indices are used for resource allocation, responsible authorities should ensure that they produce ranges reflecting the full compass of services funded. For forensic services the range of morbidity levels may be so great that funding needs to rest at a larger population level than that of health authorities. PMID- 10789352 TI - Socio-economic deprivation and the prevalence and prediction of depression in older community residents. The MRC-ALPHA Study. AB - BACKGROUND: The Townsend index is a measure of social deprivation. It can be applied to postal districts and has been employed in studies examining the ecological associations of mental illness. AIMS: We examine the utility of the Townsend index in identifying older populations with a high prevalence and risk of developing depression. METHOD: The study was carried out in the context of a cohort study of an age- and gender-stratified sample of 5222 community residents aged 65 years and over. Subjects were interviewed at intervals of two years. The relationships between Townsend score and psychiatric diagnoses (in particular, depression) were examined. RESULTS: High Townsend scores were associated with increased prevalence and incidence of depression and prevalence of organic psychiatric illness. CONCLUSIONS: The Townsend index can be used to prioritize psychiatric and primary care resources so as to cater for older populations likely to suffer from depression and organic psychiatric conditions. PMID- 10789353 TI - Post-partum depression and the mother-infant relationship in a South African peri urban settlement. AB - BACKGROUND: Post-partum depression in the developing world has received little research attention, and its association with disturbances in the mother-infant relationship is unknown. AIMS: To determine the prevalence of post-partum depression and associated disturbances in the mother-infant relationship in Khayelitsha, a South African peri-urban settlement. METHOD: The mental state of 147 women who had delivered two months previously was assessed, and the quality of their engagement with their infants was determined. RESULTS: The point prevalence of DSM-IV major depression was found to be 34.7%. Maternal depression was associated with poor emotional and practical support from the partner. It was also associated with insensitive engagement with the infants. CONCLUSIONS: The rate of post-partum depression in Khayelitsha was around three times that found in British post-partum samples, and these depressions were strongly associated with disturbances in the mother-infant relationship. PMID- 10789354 TI - Heritability of social cognitive skills in children and adolescents. AB - BACKGROUND: Social cognitive skills are those which enable understanding of social situations; they are relevant to a variety of psychiatric disorders including autism, schizophrenia and externalizing behaviour problems in children. AIMS: To examine the heritability of social cognitive skills. METHOD: Using a population-based sample of twins aged 5-17, the genetic and environmental influences on social cognitive skills were examined. RESULTS: Male scores were higher than female scores (P < 0.001), indicating poorer social cognition among males. A heritability of 0.68 (95% CI 0.43-0.78) was found, with shared environmental influences accounting for only 0.05 of the variance (95% CI 0.00 0.28). This could be removed from the model without worsening the fit. There were no significant differences in genetic effects between the genders, but age related changes were found, with younger twins showing greater genetic influence on social cognition. CONCLUSIONS: Social cognition appears to be under considerable genetic influence in the population and shows significant male female differences. No gender differences in genetic influences on the variance of scores were found, but the effects of age were significant. PMID- 10789355 TI - Relationship with Family of Origin Scale (REFAMOS). Interrater reliability and associations with childhood experiences. AB - BACKGROUND: Family relationships in the transition from childhood to adult life may be important mediators of risk and resilience for adult psychopathology. AIMS: To develop a reliable and valid measure of the quality of relationships with each parent in young adults. METHOD: Interrater reliability of the Relationship with Family of Origin Scale (REFAMOS) was assessed from audio-taped interviews with 59 subjects. Age-related trends and associations with recalled childhood relationships were examined in survivors of childhood cancer and their controls (n = 178). RESULTS: Intraclass correlation coefficients were in the range 0.69-0.95, and kappa values 0.80-0.82. Indices of current closeness to mothers were negatively correlated with age of subject and positively correlated with recalled maternal care in childhood. Negative qualities in current relationships were correlated with recalled overprotection. CONCLUSIONS: The REFAMOS has good interrater reliability, and shows the predicted age-related trends in scores and associations with recalled childhood relationships. PMID- 10789356 TI - Cognitive therapy or imaginal exposure in the treatment of post-traumatic stress disorder. Twelve-month follow-up. AB - BACKGROUND: Previously reported results have demonstrated the efficacy of exposure and cognitive therapy in the treatment of chronic post-traumatic stress disorder (PTSD), but have not shown one to be superior to the other. AIMS: To investigate whether treatment benefits and equivalence are maintained at 12-month follow-up in patients with chronic PTSD treated with either imaginal exposure or cognitive therapy. METHOD: Twelve-month follow-up of a randomised clinical trial. RESULTS: Fifty-four subjects (87% of the sample) were available to follow-up. They did not significantly differ clinically from drop-outs. There was significant clinical improvement at 12 months compared with pre-treatment. However, 39% of those followed-up still met criteria for PTSD. There were no significant differences between the two treatments. Victims of crime displayed higher levels of symptoms at follow-up than victims of accidents. CONCLUSIONS: Clinical benefits for exposure or cognitive therapy were maintained. PMID- 10789357 TI - Active monitoring of 12,760 clozapine recipients in the UK and Ireland. Beyond pharmacovigilance. AB - BACKGROUND: People prescribed clozapine for treatment-resistant schizophrenia have mandatory haematological monitoring through a case register for identifying reversible neutropenia. AIMS: To quantify risk factors for agranulocytosis in subjects receiving clozapine. METHOD: Data from 12,760 subjects registered to receive clozapine from January 1990 to April 1997 were analysed. Risk factors for agranulocytosis were quantified using a Cox proportional-hazards regression analysis. RESULTS: The risk for agranulocytosis in Asian subjects was 2.4 times that in Caucasians (P = 0.03). There was an age-related increase in risk of 53% per decade (P = 0.0001). CONCLUSIONS: The case register yielded valuable information for guiding research into the causes of the haematological reactions. PMID- 10789358 TI - Randomisation in clinical trials. AB - BACKGROUND: Several studies of papers published in non-psychiatric medical journals that report on randomised controlled trials (RCTs) indicate that there is inadequate reporting of the process by which randomisation is carried out. AIMS: To examine the adequacy of the reporting of the procedure of randomisation in clinical trials of parallel design published in the British Journal of Psychiatry (BJP) and the American Journal of Psychiatry (AJP). METHODS: All issues of the BJP and the AJP published between January 1990 and December 1998 were surveyed, and papers that reported on RCTs were examined to judge the adequacy of the reporting of the process of randomisation. RESULTS: We found 183 papers which claimed to report on RCTs (73 in the BJP and 110 in the AJP). Nine (8.2%) of those in the AJP and six (8.2%) in the BJP described the technique of creating the randomisation sequence. Two (1.8%) of those in the AJP and 11 (15.1%) of those in the BJP described the mechanism of allocating treatment. One paper in the AJP and five papers in the BJP described both the generation of random numbers and allocation. CONCLUSIONS: Adequate reporting of the method of randomisation was uncommon. The RCT status of some of the papers must therefore be in doubt. PMID- 10789359 TI - Predicting violence. PMID- 10789360 TI - Homicide and failure of community care. PMID- 10789361 TI - Suicide in farmers in India. PMID- 10789362 TI - Fever and acute brief psychosis in developing countries. PMID- 10789363 TI - People at risk of schizophrenia and other psychoses: comments on the Edinburgh High-Risk Study. PMID- 10789364 TI - Comparing ICD-10 and DSM-IV. PMID- 10789365 TI - Memory recovery among adults in therapy. PMID- 10789366 TI - Ecstasy use and neuropathology. PMID- 10789367 TI - Selective serotonin reuptake inhibitors and personality change. PMID- 10789368 TI - More on amiodarone-induced depression. PMID- 10789369 TI - Venlafaxine and paroxetine in treatment-resistant depression. PMID- 10789370 TI - Urinary detection of olanzapine--an aid to compliance. PMID- 10789371 TI - Olanzapine in the treatment of anorexia nervosa. PMID- 10789372 TI - Isolation, identification and determination of a magenta subsidiary colour in food blue no. 1 (brilliant blue FCF). AB - A magenta subsidiary colour was isolated from commercial Food Blue No. 1 (B-1; Brilliant Blue FCF). The absorption maximum for this subsidiary colour at 580 nm is outside of the range of 614-628 nm found for other subsidiary colours and m,m B-1. On the basis of MS and NMR analyses, the structure of the subsidiary colour was elucidated as the disodium salt of 2-[[4-[N-ethyl-N-(3 sulphophenylmethyl)amino]phenyl][4-oxo- 2,5 cyclohexadienylidene]methyl]benzenesulphonic acid. HPLC analyses revealed that 24 batches of commercial Food Blue No. 1 (three manufacturers) contain 0.1-0.8% (average: 0.5%) of the magenta subsidiary colour. PMID- 10789373 TI - Exposure of cadmium from infant formulas and weaning foods. AB - Cadmium levels were determined in 59 baby food samples, including milk-based, cereal and milk-based and soy-based formulas, recommended from 0 to 18 months of age. Determinations were performed by Zeeman graphite furnace atomic absorption spectrophotometry, after dry ashing, with parallel determinations of certified reference samples. Mean cadmium levels were found to range from 1.10 to 23.5 micrograms/kg fresh weight concentrated formulas. Levels were related to the composition of the diets. Formulas based on cow's milk had the lowest concentrations. Soy formulas contained approximately six times more cadmium than cow's milk formulas, and diets with a cereal content had 4-21 times higher mean levels. The mean weekly intakes of dietary cadmium were estimated to vary between 0.10 and 3.05 micrograms/kg body weight of the child, if the recommended amount of formula were to be consumed at the recommended age, and if the child were of average weight. This estimation however does not include the contribution of cadmium from drinking water. The highest intake on a body weight basis was found in 6-month-old children, consuming the recommended amount of wheat-, oat- and milk-based formulas. This intake is below the provisional tolerable weekly intake (PTWI) of 7 micrograms/kg body weight, established by a WHO/FAO expert group. However, the risk assessment is based on renal effects in adults. Furthermore, preparations of a formula by adding water with cadmium concentrations at the WHO guideline level could lead to intakes at the PTWI level. Compared to breast-fed children, the exposure of dietary cadmium from weaning diets can be up to 12 times higher in children fed infant formula. PMID- 10789374 TI - Dietary exposures to nitrate in the UK. AB - Dietary exposures to nitrate have been estimated for two different groups of UK consumers. The daily dietary exposure of the general population was estimated from the UK 1997 Total Diet study to be 52 mg/day. Vegetables contributed approximately 70% to this total dietary exposure. Dietary exposure estimates for adult consumers of vegetables commonly eaten in the UK were 93 mg/day and 140 mg/day for mean and 97.5th percentile consumers, respectively. Dietary exposures to nitrate of UK consumers are similar to those in other European countries and are less than the Acceptable Daily Intake (ADI) for nitrate set by the European Commission's Scientific Committee for Food. PMID- 10789375 TI - Screening of selected pesticides for oestrogen receptor activation in vitro. AB - Twenty pesticides were tested for their ability to activate the oestrogen receptor in vitro using an MCF7 cell proliferation assay and a Yeast Oestrogen Screen. The fungicides fenarimol, triadimefon, and triadimenol were identified as weak oestrogen receptor agonists, which at 10 microM induces a 2.0, 2.4, and 1.9 fold increase in proliferation of human MCF7 breast cancer cells (E3 clone). The relative proliferation efficiency (RPE) was 43-69%, indicating partial agonism at the oestrogen receptor. Several pesticides did not have any effect on the proliferation response after 6 days of exposure, including: chlorpyrifos, diuron, iprodion, linuron, pentachlorphenol, prochloraz, propioconazol, propyzamine, quintozen, tetrachorvinphos and tetradifon. Some pesticides resulted in a negligible proliferation response, which was not statistically significant under the present experimental conditions. These were: bromopropylate, chlorfenvinphos, chlorobenzilate, dicofol, heptachlor, and imazalil. Fenarimol and dicofol also gave rise to a positive oestrogenic response in yeast cells transfected with the oestrogen receptor alpha, whereas the remaining compounds resulted in a negative response due either to biological inactivity or cytotoxocity to the yeast cells. The EC50 for fenarimol was estimated to be 13 microM in the yeast cells, compared with an EC50 of 3 microM in the MCF7 cells, indicating higher sensitivity of the latter assay. No in vivo data for fenarimol, triadimefon or triadimenol have previously been published that support oestrogenic activity in the intact animal. Thus, from the present results we suggest that oestrogen receptor activation may not be an important mode of action for these compounds. The need to include at least two bioassays in a screening procedure and for combining in vitro and in vivo data is emphasized. PMID- 10789376 TI - Evaluation of a single plate microbiological growth inhibition assay as a screening test for the presence of antimicrobial agents in compound animal feedingstuffs at therapeutic and contaminating concentrations. AB - The Inhibitory Substance Test (IST), a microbiological growth inhibition test, is used for screening animal feedingstuffs for the presence of (contaminating) antimicrobial compounds. The effectiveness of the IST was established for 33 compounds that may be incorporated in feedingstuffs. Minimum detectable concentrations (MDCs) for standard solutions were established and compared with those obtained following solvent extraction of an antimicrobial-free compound feedingstuff spiked with each compound at 0-20 mg/kg. Of the 33 standard solutions examined, the test organism was not sensitive to 11 and the MDC for one was greater than its maximum inclusion rate in complete feedingstuffs. Following routine extraction (25% acetone-phosphate buffer) of feedingstuffs spiked with each of the 22 compounds to which the organism was sensitive, 10 were not detected, 15 were detectable at both minimum and maximum feed-inclusion rates and four were only detectable at their maximum feed-inclusion rates. Extraction with methanol (25%) had a deleterious effect with 12 compounds not detected, nine detectable at both minimum and maximum feed-inclusion rates and five detectable at their maximum feed-inclusion rates. Increasing acetone and methanol concentrations to 40 and 55% respectively resulted in larger inhibitory zones for antibiotic-free feedingstuff (25.3 + 2.43 mm vs 21.1 + 1.02 mm) compared with both 25% acetone (11.3 + 0.22 mm) and 25% methanol (11.2 + 0.22 mm), requiring the establishment of greater threshold zone diameters and negating any advantage in increasing the solvent concentration under these test conditions. It is concluded that the IST may be particularly useful for detection of a number of the zootechnical feed-additives recently banned in the EU, which, if used illegally, may be present at sufficiently high inclusion rates to facilitate detection. Further alteration of extraction conditions may improve the scope of the assay. PMID- 10789377 TI - Control of growth and fumonisin B1 production by Fusarium verticillioides and Fusarium proliferatum isolates in moist maize with propionate preservatives. AB - The effect of propionic acid, its sodium salt or a commercial formulation of propionates (0.03, 0.05 and 0.07%), on growth and fumonisin B1 production by Fusarium verticillioides and F. proliferatum isolated was evaluated on irradiated maize at different water activities (aw, 0.93, 0.95, 0.98) and temperatures (15, 25 degrees C). The four isolates grew at all aw x temperature treatments in the absence of propionates. At the highest propionate concentration tested (0.07%), however, growth was restricted to 0.98 aw, for F. proliferatum isolates but not for those of F. verticillioides. Inhibition of growth was maximum when propionates were added in the acid form. In the presence of low propionate concentrations (0.03%), growth was sometimes enhanced probably due to assimilation of these compounds by the fungus. Water activity, temperature, concentration and source of propionate, as well as most two-, three-, four-, and five-way interactions had a significant influence on growth of Fusarium isolates. None of the assayed treatments had any effect on fumonisin B1 production by F. verticillioides isolates. For F. proliferatum, higher fumonisin B1 production occurred in the absence of propionates, and in general concentration decreased with increasing doses of preservatives. Single factors (aw, propionate concentrations and temperature) and temperature x aw and propionate concentration x temperature interactions had a significant effect on fumonisin production (p < 0.01). Moreover, propionate concentration was the single most important factor, besides temperature, which affected fumonisin B1 production. PMID- 10789378 TI - Natural co-occurrence of fumonisins, deoxynivalenol, zearalenone and aflatoxins in field trial corn in Argentina. AB - Corn samples collected from the main production area in Argentina in 1995 were surveyed for the natural occurrence of Fusarium mycotoxins and aflatoxins. Fumonisins B1, B2 and B3 and zearalenone were found in all samples. A positive relationship was found between fumonisins B1, B2 and B3, B1 and B3, and B2 and B3. Deoxynivalenol and aflatoxins were not detected. Mycological survey has also revealed the predominance of Fusarium moniliforme. This is the first report on the simultaneous occurrence of fumonisins and zearalenone in corn from the main production area in Argentina. PMID- 10789379 TI - Phthalates in paper and board packaging and their migration into Tenax and sugar. AB - Packaging samples for many kinds of foodstuffs were received from manufacturers together with basic information about the materials used in their production. Half of the 29 samples studied contained phthalates in amounts exceeding 5 mg/kg. Two types of paper bag intended for sugar and exceptionally high phthalate contents although they were flexo printed. The maximum contents of di isobutylphthalate (DIBP) and dibutylphthalate (DBP) were 450 and 200 mg/kg, respectively. The phthalates found originated from adhesives used in the joints of the packaging. In other packagings manufactured at the same time, DIBP concentrations varied from 92 to 193 mg/kg. Phthalates were also determined in sugar before and after packaging. Migration of phthalates ranged from 57 to 74% of the original content in the packaging after 4 months storage. Packed sugar contained DIBP 2.2-2.6 mg/kg and DBP 0.5-1 mg/kg. The sugar packagings were also tested using Tenax as a food stimulant. The results indicated that 69-91% of the original content of the phthalates migrated into Tenex after 10 days at 40 degrees C. The major drawback with migration testing using Tenax is its high cost; it is not practical to fill the whole 1 kg packaging with Tenax, and smaller samples of packaging must be used instead. However, the distribution of substances in the packaging might be non-uniform. In the sugar packagings studied here, the difference between phthalate concentrations in two samples taken from the same packaging was nearly 100-fold. PMID- 10789380 TI - The migration from the internal coatings of food cans; summary of the findings and call for more effective regulation of polymers in contact with foods: a review. AB - The analysis of migrates from the internal coatings of food cans showed that the present regulation is not sufficient to ensure safety of food contact polymers. The migrates often consist of complex mixtures of reaction products and impurities, and their harmlessness cannot be deduced from the use of non-toxic starting substances compiled in positive lists. The regulation proposed herein asks producers to analyse the composition of the migrates. It includes an upper limit for the molecular weight (e.g. 1000 Da) and thresholds defining concentrations in the foods down to which components must be identified and evaluated. Primarily considering feasibility at reasonable costs, thresholds of 30 micrograms/kg for individual components and 300 micrograms/kg for the sum of the unidentified materials are proposed. Producers must be able to demonstrate that the individual components or mixtures exceeding this limit, including reaction products with food components, have no toxic effect. PMID- 10789381 TI - Alcohol: matters of life and death. PMID- 10789382 TI - Self-regulation in the independent sector. PMID- 10789383 TI - Photodynamic therapy. PMID- 10789384 TI - Prevention and treatment of malaria in UK travellers. AB - With around 2500 new cases of malaria being reported each year and more than a million UK residents visiting malaria endemic countries, there has never been a greater need for effective prevention and treatment. The latest information on malaria management is outlined below. PMID- 10789385 TI - Travel and the immunocompromised host. AB - Increasing opportunities for travel and advances in medicine mean that immunocompromised patients may venture to potentially risky parts of the world. This article examines the risks faced by such travellers. Some limitations of standard travel vaccines are discussed and suggestions are made as to how best to advise such travellers. PMID- 10789386 TI - Tuberculosis and travel. AB - Tuberculosis is increasing worldwide with most countries having a high prevalence. Protection against tuberculosis, the risks of acquiring tuberculosis during air travel, and the evidence for clinical tuberculosis acquired on extended visits to such countries are all reviewed in this article. PMID- 10789387 TI - Challenging the stigma of Alzheimer's disease. AB - Changing Minds--Every Family in the Land is a national campaign launched by the Royal College of Psychiatrists which aims to reduce discrimination against people who suffer from mental disorders. One of its target areas is dementia. What is the stigma associated with Alzheimer's disease and how can hospital doctors challenge it in everyday practice? PMID- 10789388 TI - Chest pain in the emergency department. AB - The assessment of patients with chest pain is challenging for any emergency physician because of the spectrum of illness covered by this symptom. Patients may have a serious life-threatening condition or a trivial self-limiting illness. This article presents an approach to the assessment and early management of patients presenting with acute non-traumatic chest pain in the emergency department. PMID- 10789390 TI - Recovery from surgery: grappling with an elusive concept. AB - When we ask our patients 'How are you?', can we quantify the answer? There is no ideal measure for every circumstance. This article will discuss the issues in measuring postsurgical recovery, describe different approaches, and consider future research. PMID- 10789389 TI - The thiazolidinediones: a new class of antidiabetic agents. AB - Insulin resistance is a fundamental feature of type 2 diabetes and is strongly associated with metabolic disorders which predict increased cardiovascular risk, including hypertension and dyslipidaemia. A new class of insulin-sensitizing agents, the thiazolidinediones, reduce insulin resistance and improve glycaemia both as monotherapy and in combination with sulphonylureas or metformin. PMID- 10789391 TI - Clinical advances in the evaluation of oesophageal disease. AB - The rising incidence of gastro-oesophageal reflux disease and its sequelae means the evaluation of oesophageal disease is set to expand beyond routine endoscopic assessment. The techniques used in the evaluation of both benign and malignant oesophageal disease are outlined in this article. PMID- 10789392 TI - Importance of components of postgraduate education in obstetrics and gynaecology: surveys of trainees and trainers. AB - Postgraduate medical education has many elements. Some are long-established while others such as formal teaching programmes and computer-aided learning are relatively new. The opinions of trainees and trainers differ as to which elements have the most educational value. Information technology is not highly rated by either group. PMID- 10789393 TI - Is childbirth safe in the UK and are there enough obstetricians? Letter to a chief executive. AB - If you are at term after a normal pregnancy and are not lucky enough to live in Portsmouth, then your chance of losing the baby during labour is higher in Great Britain than in nine other European countries, including Slovenia. The NHS Litigation Authority have put aside 1.3 billion Pounds for the next 5 years to cover litigation relating to cases of cerebral palsy, the majority of which will occur in GP- and midwife-led units. The European work directive will severely restrict staff grade and consultant hours of work over the next few years and this can only be made up by appointing more consultants. PMID- 10789394 TI - New medicine for the new millennium: a time for change? AB - Health care is under scrutiny. Demand exceeds supply and funding is insufficient. Rationing is a reality while providers feel persecuted. Yet patients remain patient--and continue to suffer. What can be done to improve the present situation? PMID- 10789395 TI - The demon drink. PMID- 10789396 TI - MADD or drunk? Adults have inborn errors too. PMID- 10789397 TI - Hypertension secondary to Marfan's syndrome initially attributed to an anxiety disorder. PMID- 10789398 TI - Putting the brakes on child traffic casualties. PMID- 10789399 TI - Violence towards doctors in Turkey. PMID- 10789400 TI - Praise for flexible training. PMID- 10789402 TI - Jargon busters: individual savings accounts. PMID- 10789401 TI - Web watch. PMID- 10789403 TI - Current issue has changed in several subtle ways. PMID- 10789404 TI - Hearing: cracking the code. PMID- 10789405 TI - Metastases from glomus jugulare tumours. AB - This review describes the features of glomus jugulare tumours with metastases. There were 100 sites of metastasis in the 53 cases previously reported. The sites of metastasis may be summarized as bone (33), lungs (23), lymph nodes (19), liver (nine) and other (16). Metastases presented up to 30 years after the initial treatment. The mean age of patients was 45 years and the sex ratio was approximately two females to one male, with no significant difference compared to non-metastatic tumours. There was a significantly higher incidence of pain and a significantly lower incidence of hearing loss at presentation compared to non metastatic tumours. The commonest treatment was a combination of surgery and radiotherapy. The duration of symptoms before diagnosis was significantly shorter and the rates of persistent or recurrent local disease and death were significantly higher than for non-metastatic tumours. This review highlights the fact that glomus jugulare tumours are not always benign. PMID- 10789406 TI - Accuracy of patient self-medication with topical eardrops. AB - Topical eardrops are commonly prescribed by otolaryngologists and general practitioners and many believe that patient compliance is poor, although this has not been studied in a properly controlled manner. Thirty-nine patients with otitis externa were reviewed four times during a two-week course of topical eardrops and their treatment compliance studied. The patients tended to undermedicate themselves, and this tendency increased over the two-week period. Patients were administering significantly fewer drops by the end of day 11 than by the end of day 3 (p = 0.2). Compliance patterns were extremely poor, in that only 40 per cent of patients were managing to self-medicate within a 25 per cent error margin by the end of day 3. These findings may have future implications in the ongoing debate on eardrop use. PMID- 10789407 TI - Middle-ear disease in remote Aboriginal Australia: a field assessment of surgical outcomes. AB - Chronic middle-ear disease is highly prevalent among Australian Aboriginal people, and many undergo surgical treatment. However, the outcomes of surgery in this group have not been fully evaluated. This is a descriptive study of operations for middle-ear disease (excluding grommets) on Aboriginal patients in Kimberley hospitals between 1 October 1986 and 31 December 1995. Logistic regression was used to model predictors of surgical outcome. Success was defined by an intact tympanic membrane and air-bone gap of < or = 25 dB at review at, or later than, six months post-operation. A success rate of 53 per cent was observed; increasing age was the only variable predictive of success. Successful outcomes were more likely in adults and children aged > 10 years, however, this does not take into account the necessity of hearing for language acquisition and learning. Dedicated resources must be allocated for post-operative follow-up of Aboriginal patients so that much-needed, rigorous evaluations of ENT surgery can be conducted. PMID- 10789408 TI - Performance after cochlear implantation: a comparison of children deafened by meningitis and congenitally deaf children. AB - The speech perception and speech production performance following cochlear implantation of congenitally deaf children and children deafened by meningitis were analysed. Three groups consisting of 70 congenitally deaf children, 22 children deafened by meningitis before two years of age and 14 children deafened by meningitis after two years of age were compared. The group deafened by meningitis after two years of age demonstrated significantly better speech perception than the other two groups. Their speech production appeared better but did not achieve statistical significance compared with the other two groups. There was no significant difference in either speech perception or speech production between the congenitally deaf group and the group deafened by meningitis before two years of age. PMID- 10789409 TI - Routine coagulation screening in the management of emergency admission for epistaxis--is it necessary? AB - The role of routine coagulation studies in the management of patients suffering from epistaxis is unclear. In an attempt to address this issue the case notes of all emergency admissions for epistaxis to a large Scottish teaching hospital were retrospectively reviewed over a one-year period. One hundred and forty patients (63 male, 77 female) were admitted between January and December 1998. The patients who had coagulation studies were identified and their results analysed. A total of 121 patients (86.4 per cent) had coagulation studies performed. Of these, 10 (8.3 per cent) had abnormal results and all were taking warfarin or a combination of warfarin and aspirin. No other coagulation abnormalities were identified. This study supports the view that there does not appear to be a role for routine coagulation studies in patients admitted with epistaxis. The investigation for potential haemostatic disorders should be performed when clinically indicated and, if necessary, in consultation with the haematology service. PMID- 10789410 TI - Identification and typing of human papillomavirus (HPV) in squamous cell carcinoma of the oral cavity and oropharynx. AB - Recent epidemiological research has raised the possibility of a connection between viral infection by the human papilloma virus (HPV) and the existence of squamous cell carcinoma of the oral cavity and oroharynx (SCCa). Some 60 subtypes have been identified to date, and specific subtypes are associated with lesions or infections at specific tissue sites. Twenty-three patients with SCCa of the oral cavity were studied for the presence of HPV in their tumours. HPV DNA (all type 16) was isolated from four specimens (17.3 per cent) using the Polymerase Chain Reaction (PCR) method. The four patients were all over age 50, and had advanced tumours (two T3 and two T4) that originated in the oropharynx. Our data supports other reports in the literature, demonstrating the presence of HPV DNA type 16 within cells from oropharyngeal SCCa. We conclude that the PCR method is a useful and reliable method for the detection of HPV DNA within tumour cells. PMID- 10789411 TI - Labyrinthine involvement and multiple perforations of the tympanic membrane in acute otitis media due to group A streptococci. AB - We present here three cases of acute otitis media caused by a virulent group A streptococcal infection that rapidly led to deterioration in hearing. Two of the three cases presented with severe sensorineural and mixed hearing loss with multiple tympanic membrane perforations, and the third presented with severe bilateral sensorineural hearing loss following acute otitis media involving group A streptococci. All patients were treated with systemic (piperacillin) and topical antibiotics (ofloxacin ear drops): one patient also received a systemic steroid (betamethasone). Deafness persisted in one patient but in the other two, hearing gradually recovered. Severe cytotoxicity was considered to have occurred in all patients, resulting in multiple perforations of the tympanic membrane and necrosis in the middle ear. PMID- 10789412 TI - Mould constituents in the middle ear, a hearing-aid complication. AB - During the placement of a mould for a hearing-aid by a hearing-aid dispenser, moulding material entered the middle ear through pre-existent perforations in the tympanic membrane in both ears. Besides hearing loss, there were no other symptoms. Surgical removal of the moulding material by tympanotomy was necessary, and was complicated by encirclement of the ossicles by the material. All the material could be removed and the hearing was saved. Recommendations for an improved procedure of mould-making are made including more detailed information of the otoscopic findings at the prescriptions for hearing-aid moulds. PMID- 10789413 TI - A case of extramedullary plasmacytoma arising from the nasal septum. AB - A rare case of extramedullary plasmacytoma arising from the nasal septum with localized amyloid deposition is reported. A 75-year-old woman presented with a history of post-nasal discharge and nasal obstruction for several months. Endonasal endoscopic observation revealed the presence of a mass arising from her nasal septum with an extension to the nasopharynx. No cervical lymph nodes were palpable. The biopsy specimen was diagnosed as a plasmacytoma (IgG, lambda-light chain type). Both serum myeloma-protein and urine Bence-Jones protein were negative. Bone marrow biopsy, a chest radiograph, total body skeletal survey and 67Ga- and 99mTc-scintigrams showed no other systemic lesions. These findings confirmed the diagnosis of extramedullary plasmacytoma in the nasal septum. The patient received irradiation of 40 Gy without clinically detectable reduction of tumour size. The patient eventually underwent complete resection of the tumour by KTP/532 laser under endonasal endoscopic control. Pathologically, the tumour mass was composed mainly of amyloid deposition with a marked reduction of tumour cells. This indicated the radiosensitiveness of tumour cells, which was clinically masked by the increased amyloid deposition. The clinical presentation, pathological features and surgical procedures are described with a review of the literature. PMID- 10789414 TI - Living on the edge of the post-nasal space: the inhaled foreign body. AB - We present a previously undescribed event where an inhaled foreign body was propelled into the post-nasal space by the Heimlich manoeuvre. We present figures showing a drop in the mortality following inhaled foreign bodies since the introduction of the Heimlich manoeuvre. PMID- 10789415 TI - Surgical approach to a giant fibrolipoma of the supraglottic larynx. AB - This paper describes the surgical procedures for a fibrolipoma that first appeared as a giant tumour in the hypopharynx and extended to the cardiac antrum of the oesophagus. At the initial surgery, a pedunculated tumour originating from the left arytenoid of the larynx was found to occupy the cervical as well as thoracic oesophagus and was thus removed through a lateral pharyngectomy. A histological examination revealed fibrolipoma. However there was a recurrence of the tumour in the arytenoid and the patient suffered from dysponea. In addition, a submucosal tumour was also found in the left false vocal fold. At the second surgery, the masses in the arytenoid and false vocal fold were subtotally removed without damaging the mucosa. The mucosa of the arytenoid was sutured to the thyropharyngeal muscle on the same side and the arytenoid swelling disappeared almost completely. The post-operative course has been uneventful for more than two years. PMID- 10789416 TI - Hoarse voice resulting from premature ageing in Werner's syndrome. AB - Werner's syndrome is characterized by clinical signs of premature ageing. A 42 year-old man presented with three-year history of hoarseness. Also noted were skin atrophy of the face and hands, ulcerations around the ankles, and a history of cataracts. A clinical diagnosis of Werner's syndrome was made. Laryngoscopy revealed bowed vocal folds resulting in a spindle-shaped defect with glottal incompetence during phonation. Examination also revealed decreased maximum phonation time and vocal fatigue. At surgery, atrophy of the vocalis muscle was noted. Furthermore, degeneration of muscle fibres was noted in the temporalis muscle. The atrophic changes in the vocal folds that occur with ageing and result in an increased fundamental frequency were seen in this patient. The characteristic hoarseness of Werner's syndrome appears to be the result of premature ageing of the vocal-folds. PMID- 10789417 TI - Primary tracheal leiomyosarcoma. AB - We present the clinical course of a 56-year-old female patient with a primary tracheal leiomyosarcoma. The diagnostic approach and pathological classification of this seldom described tumour remains extremely difficult. We discuss the symptoms as well as the diagnostic and therapeutic procedures, including multimodal chemotherapy with organ-preserving surgery leading to complete remission. PMID- 10789418 TI - Glomus jugulare tumour with metastases to cervical lymph nodes. AB - Glomus jugulare tumours are classically described as benign tumours with a long time course often measured in decades. Although these tumours may be locally invasive, most cases are histologically benign and metastases are rare. The case of a malignant glomus jugulare tumour with a particularly aggressive pattern of spread is presented. At the time of surgery, which was within 12 months of the development of symptoms, intracranial spread and metastasis to cervical lymph nodes had already occurred, demonstrating that glomus jugulare tumours are not always benign. PMID- 10789419 TI - Glossopharyngeal neuralgia following foreign body impaction in the neck. AB - Glossopharyngeal neuralgia is rare, typically idiopathic and treated with carbamazepine. Surgery to decompress or transect the glossopharyngeal nerve root may be performed if conservative management fails. We present a case following trauma to the neck with foreign body impaction. To our knowledge this is the first case of glossopharyngeal neuralgia due to neck trauma. PMID- 10789420 TI - Extraskeletal Ewing's sarcoma of the scalp. AB - Extraskeletal Ewing's sarcoma/primitive neuroectodermal tumour (EES/PNET) is a rare disease entity. Scalp EES/PNET has been reported rarely. We report a case of an 11-year-old boy who had painful and rapidly growing subcutaneous nodes over the scalp and neck. The final diagnosis was EES/PNET after biopsy and immunohistochemical assay. The patient underwent surgical excision, chemotherapy and radiotherapy with a dose of 2000 cGy. Now he has been free of disease for two years. Early awareness and treatment of this rare disease, and wide resection followed by chemotherapy and radiotherapy might improve patients' long-term survival. PMID- 10789421 TI - Fungal rhinosinusitis. AB - Fungal infections in both their invasive and non-invasive forms can prove difficult to diagnose. The often characteristic appearances on imaging are of great assistance. CT is the primary imaging modality and is probably more accurate than MRI in diagnostic specificity and determining the extent of bone erosion. However this may require a modified scanning technique to adequately demonstrate the typical soft tissue density variations of fungi. MRI should be used to supplement CT when intra-cranial or intra-orbital extension is suspected. PMID- 10789422 TI - Schwannoma of the chorda tympani nerve. AB - We report a case of schwannoma of the chorda tympani. This is a very rare benign tumour and only five other cases have been found in the literature. This is the first case to mimic a cholesteatoma presenting as a pearly tumour in the postero superior segment of the drum with aural discharge and conductive deafness. Diagnosis is usually by biopsy and treatment is surgical with preservation of facial and auditory function. A summary of the other presentations of this tumour together with a review of the histopathology of the disease is presented. PMID- 10789423 TI - Bilateral basal-cell adenomas in the parotid glands. AB - A rare case of bilateral basal-cell adenomas in the parotid glands of a 65-year old woman is reported. There have been only four previous reported cases of bilateral parotid basal-cell adenomas, all of which occurred synchronously with dermal cylindromas. The present lesion is the first case reported in the English literature of bilateral parotid basal-cell adenoma without coexisting dermal cylindroma. Histological examination revealed that the left tumour had adenoid cystic change, which closely resembled adenoid cystic carcinoma. This case is of interest not only because of the raity of bilateral parotid basal-cell adenomas, but also because of the necessity for differentiation from adenoid cystic carcinoma in order to plan appropriate treatment. PMID- 10789424 TI - Speed tuning of motion segmentation and discrimination. AB - Motion transparency requires that the visual system distinguish different motion vectors and selectively integrate similar motion vectors over space into the perception of multiple surfaces moving through or over each other. Using large field (7 degrees x 7 degrees) displays containing two populations of random-dots moving in the same (horizontal) direction but at different speeds, we examined speed-based segmentation by measuring the speed difference above which observers can perceive two moving surfaces. We systematically investigated this 'speed segmentation' threshold as a function of speed and stimulus duration, and found that it increases sharply for speeds above approximately 8 degrees/s. In addition, speed-segmentation thresholds decrease with stimulus duration out to approximately 200 ms. In contrast, under matched conditions, speed-discrimination thresholds stay low at least out to 16 degrees/s and decrease with increasing stimulus duration at a faster rate than for speed segmentation. Thus, motion segmentation and motion discrimination exhibit different speed selectivity and different temporal integration characteristics. Results are discussed in terms of the speed preferences of different neuronal populations within the primate visual cortex. PMID- 10789426 TI - Adaptation to temporal modulation can enhance differential speed sensitivity. AB - During adaptation to a moving pattern, perceived speed decreases. Thus we know that the adapted visual system does not simply code the absolute speed of a stimulus. We hypothesised that adaptation to a moving stimulus serves to optimise coding of changes in speed at the expense of maintaining an accurate representation of absolute speed. In this case we would expect discrimination of speeds around the adapted level to be preserved or enhanced by motion adaptation. Speed discrimination thresholds were measured for sinusoidal gratings (1.25 cpd; 12.5 Hz; 40% contrast) with and without prior adaptation to moving, static, and flickering stimuli. After adaptation to motion in the same direction as the test, seven of eight subjects showed a reduction of perceived speed in the adapted region, and seven showed enhanced discrimination. Similar effects were found for adaptation to motion in the opposite direction to the test and to counter-phase flicker, suggesting that adaptation is driven by temporal modulation rather than by motion per se. We conclude that motion adaptation preserves or enhances differential speed sensitivity at the expense of an accurate representation of absolute speed. PMID- 10789427 TI - Depth aliasing by the transient-stereopsis system. AB - A fundamental problem in stereo-processing is determining which images in the two eyes correspond to the same object. This problem is particularly pronounced with periodic stimuli where it is theoretically possible to binocularly match a given feature in one eye with any of the identical features in the other eye. One way to minimise the likelihood of the occurrence of such aliasing is to restrict the upper-disparity limit that a particular binocular cell can process to one-half of the spatial period to which the cell is sensitive. While such a restriction would not be a major problem for the sustained stereo-system (which processes small disparities) it would be for the transient system (which is capable of processing disparities as large as 10 degrees). Large-field sinewave variations in luminance were used to compare the propensity of the sustained and transient systems to exhibit depth aliasing--that is to signal a depth sign that corresponds to a binocular match that is greater than the nearest-neighbour pairing. Results were that: depth aliasing was exhibited at short, but not at long durations; decreasing the disparity of the stimulus reduced the likelihood of depth aliasing; and the critical disparity for this reduction in depth aliasing was dependent upon the spatial frequency of the stimulus, i.e. it was phase, not absolute disparity dependent. Based upon these results, we conclude that while the sustained system implements the half-cycle disparity-processing limit, the transient system does not. PMID- 10789425 TI - A new approach to the study of ocular chromatic aberrations. AB - We measured the ocular wavefront aberration at six different visible wavelengths (between 450 and 650 nm) in three subjects, using a spatially resolved refractometer. In this technique, the angular deviation of light rays entering the pupil at different locations is measured with respect to a target viewed through a centered pupil. Fits of the data at each wavelength to Zernike polynomials were used to estimate the change of defocus with wavelength (longitudinal chromatic aberration, LCA) and the wavelength-dependence of the ocular aberrations. Measured LCA was in good agreement with the literature. In most cases the wavefront aberration increased slightly with wavelength. The angular deviations from the reference stimulus measured using a magenta filter allowed us to estimate the achromatic axis and both optical and perceived transverse chromatic aberration (TCA), (including the effect of aberrations and Stiles-Crawford effect). The amount of TCA varied markedly across subjects, and between eyes of the same subject. Finally, we used the results from these experiments to compute the image quality of the eye in polychromatic light. PMID- 10789428 TI - Grouping visual features during binocular rivalry. AB - During binocular rivalry, portions of one eye's view may be perceptually dominant while other portions are suppressed; at any given moment, overall dominance often resembles a patchwork mixture of the two eyes' views. This study investigates the potency of two Gestalt grouping cues--good continuation and common fate--to promote synchronous fluctuations in dominance of two, spatially separated rival targets. Two grating patches were presented to the left eye paired dichoptically with random-dot patches presented to corresponding right eye locations. The orientations of the two gratings were either collinear, parallel or orthogonal. Gratings underwent contrast modulations that were either correlated (identical contrast changes) or uncorrelated (independent contrast changes). Over 60 s trials, observers pressed one key when the left grating predominated, another when the right grating predominated and both keys when both were concurrently visible. Correlated contrast modulation promoted joint grating predominance relative to the uncorrelated conditions, an effect strongest for collinear gratings. Joint predominance depended strongly on the angular separation between gratings and the temporal phase-lag in contrast modulations. These findings may reflect neural interactions subserved by lateral connections between cortical hypercolumns. PMID- 10789429 TI - Are judgements of circularity local or global? AB - We assessed, in a task where subjects had to detect smooth deviations from circularity, whether the underlying mechanisms were localised in space to the size of the individual perturbations or whether they computed global shape. By manipulating the phase, the number of cycles of modulation and the spatial arrangement of the perturbations we argue that although either aspect can be detected, performance is ultimately limited by a global shape detecting mechanism. We show that this global mechanism receives input from spatially coarse, crossed orientationally tuned filters whose peak position in orientation depends on the overall shape to be detected. PMID- 10789430 TI - A multiscale spatial filtering account of the White effect, simultaneous brightness contrast and grating induction. AB - Blakeslee and McCourt ((1997) Vision Research, 37, 2849-2869) demonstrated that a multiscale array of two-dimensional difference-of-Gaussian (DOG) filters provided a simple but powerful model for explaining a number of seemingly complex features of grating induction (GI), while simultaneously encompassing salient features of brightness induction in simultaneous brightness contrast (SBC), brightness assimilation and Hermann Grid stimuli. The DOG model (and isotropic contrast models in general) cannot, however, account for another important group of brightness effects which includes the White effect (White (1979) Perception, 8, 413-416) and the demonstrations of Todorovic ((1997) Perception, 26, 379-395). This paper introduces an oriented DOG (ODOG) model which differs from the DOG model in that the filters are anisotropic and their outputs are pooled nonlinearly. The ODOG model qualitatively predicts the appearance of the test patches in the White effect, the Todorovic demonstration, GI and SBC, while quantitatively predicting the relative magnitudes of these brightness effects as measured psychophysically using brightness matching. The model also accounts for both the smooth transition in test patch brightness seen in the White effect (White & White (1985) Vision Research, 25, 1331-1335) when the relative phase of the test patch is varied relative to the inducing grating, and for the spatial variation of brightness across the test patch as measured using point-by-point brightness matching. Finally, the model predicts intensive aspects of brightness induction measured in a series of Todorovic stimuli as the arms of the test crosses are lengthened (Pessoa, Baratoff, Neumann & Todorokov (1998) Investigative Ophthalmology and Visual Science, Supplement, 39, S159), but fails in one condition. Although it is concluded that higher-level perceptual grouping factors may play a role in determining brightness in this instance, in general the psychophysical results and ODOG modeling argue strongly that the induced brightness phenomena of SBC, GI, the White effect and the Todorovic demonstration, primarily reflect early-stage cortical filtering operations in the visual system. PMID- 10789431 TI - Evidence for curvilinear interpolation from dot alignment judgements. AB - Visual interpolation between dots responsible for rectilinear versus curvilinear contour interpretation was examined with the psychophysical forced directional response (FDR) paradigm. Regular four-dot polygon segments, together with a target dot, were presented to the subjects for 150 ms. Subjects were required to indicate the direction of deviation of the target dot from the midpoint of the intermediate line segment. Crucial variables were the outer angle of the line segments and symmetry axis orientation of the polygon segment. Logistic regression analyses showed that curvilinear interpolation occurred for angles up to 30 degrees, but emerged more pervasively under the vertical symmetry axis orientation for angles up to 60 degrees. PMID- 10789432 TI - Motion contrast: a new metric for direction discrimination. AB - The Adelson-Bergen energy model (Adelson, E. H., & Bergen, J. R. (1985). Spatiotemporal energy models for the perception of motion. Journal of the Optical Society of America A, 2, 284-299) is a standard framework for understanding first order motion processing. The opponent energy for a given input is calculated by subtracting one directional energy measure (EL) from its opposite (ER), and its sign indicates the direction of motion of the input. Our observers viewed a dynamic sequence of gratings (1 c/deg) equivalent to the sum of two gratings moving in opposite directions with different contrasts. The ratio of contrasts was varied across trials. We found that opponent energy was a very poor predictor of direction discrimination performance. Heeger (1992). Normalization of cell responses in cat striate cortex. Visual Neuroscience, 9, 181-197) has suggested that divisive inhibition amongst striate cells requires a contrast gain control in the energy model. A new metric can be formulated in the spirit of Heeger's model by normalising the opponent energy (EL - ER) with flicker energy, the sum of the directional motion energies (EL + ER). This new measure, motion contrast (EL - ER)/(EL + ER), was found to be a good predictor of direction discrimination performance over a wide range of contrast levels, but opponent energy was not. Discrimination thresholds expressed as motion contrast were around 0.5 +/- 0.1 for the sampled drifting gratings used in our experiments. We show that the dependence on motion contrast, and the threshold of about 0.5, can be predicted by a modified opponent energy model based on current knowledge of the response functions and response variance of cortical cells. PMID- 10789433 TI - Eye movement control in reading: accounting for initial fixation locations and refixations within the E-Z Reader model. AB - Reilly and O'Regan (1998, Vision Research, 38, 303-317) used computer simulations to evaluate how well several different word-targeting strategies could account for results which show that the distributions of fixation locations in reading are systematically related to low-level oculomotor variables, such as saccade distance and launch site [McConkie, Kerr, Reddix & Zola, (1988). Vision Research, 28, 1107-1118]. Their simulation results suggested that fixation locations are primarily determined by word length information, and that the processing of language, such as the identification of words, plays only a minimal role in deciding where to move the eyes. This claim appears to be problematic for our model of eye movement control in reading, E-Z Reader [Rayner, Reichle & Pollatsek (1998). Eye movement control in reading: an overview and model. In G. Underwood, Eye guidance in reading and scene perception (pp. 243-268). Oxford, UK: Elsevier; Reichle, Pollatsek, Fisher & Rayner (1998). Psychological Review, 105, 125-157], because it assumes that lexical access is the engine that drives the eyes forward during reading. However, we show that a newer version of E-Z Reader which still assumes that lexical access is the engine driving eye movements also predicts the locations of fixations and within-word refixations, and therefore provides a viable framework for understanding how both linguistic and oculomotor variables affect eye movements in reading. PMID- 10789434 TI - Elicited sequential presentation for low vision reading. AB - This paper reports on a variant of the rapid serial visual presentation (RSVP) technique for low vision reading called elicited sequential presentation (ESP). In both techniques, words are presented sequentially at a constant screen location, but with ESP, the reader elicits presentation of each new word by means of a button press, rather than (as with RSVP) being presented with it automatically at fixed intervals. An experiment comparing reading speeds using, ESP, RSVP and a conventional closed-circuit television (CCTV) reading aid showed that for 15 slow readers who were customary CCTV users with low vision, ESP is superior to RSVP and yields reading speeds averaging 47% faster than RSVP--about the same as CCTV reading speed. The log of the ratio of ESP to RSVP reading speeds was significantly negatively correlated with the log of RSVP reading speed, showing that slower readers benefit more than faster readers; regression predicted no benefit for readers who read with RSVP at 133 wpm or greater. Finally, word length and word presentation duration chosen by subjects reading with ESP were significantly correlated, suggesting that part of the benefit of ESP is due to reader's ability to allocate time based on word length and difficulty. PMID- 10789435 TI - Ferrocenyl derivatives with one, two, or three sulfur-containing arms for self assembled monolayer formation AB - Self-assembled monolayers of electroactive molecules can form on gold electrodes if the molecules include a sulfur-containing group to coordinate with the gold surface. We have prepared a molecule with a tripod of sulfur groups that has the potential of fixing the geometry of the molecule relative to the gold surface. The target (3) contained the good one-electron donor ferrocene connected through a benzene spacer to an isobutane tripod, with each arm of the tripod ending in a methylthio group. Analogous compounds with one (1) and two (2) coordinating arms were also prepared. PMID- 10789436 TI - Additions to Fe(CO)3-dienal complexes: dependence of diastereoselectivity on Lewis acid. AB - Lewis acid mediated additions to Fe(CO)3-2,4-dienal complexes are explored. Silyl enol ether and allylstannane nucleophiles both undergo aldehyde addition with good diastereoselectivity. However, with TiCl4 as the catalyst, the psi-exo diastereomer is the major product while with BF3.Et2O the psi-endo diastereomer is favored. Lewis acid/substrate interactions are studied by VT-NMR, and a hypothesis that accounts for the observed diastereoselectivity is presented. PMID- 10789437 TI - Unusual regioselectivity of Pd(0)-catalyzed coupling reaction of electron deficient alkenyl halides with allenic/propargylic zinc reagents. Highly selective synthesis of 6-phenylhex-5-yn-2(or 3)-enoates/enitrile and 4-phenyl-6 substituted-hexa-2,4,5-trienoates AB - The Pd(0)-catalyzed coupling reaction of electron-deficient alkenyl halides with the organozinc formed by the subsequent treatment of 1-phenylalk-1-yne with n BuLi and ZnBr2 with or without a catalytic amount of HgCl2 was studied. Both the allene-formation- and the alkyne-formation-type coupling reactions were observed with high regio- and stereoselectivity: the reaction of 1-phenylprop-1-yne afforded 6-phenylhex-5-yn-2(or 3)-enoates and -enitriles, while the reaction of 1 phenylhex-1-yne formed 4-phenyl-6-substituted-hexa-2,4,5-trienoates. A plausible explanation for the regioselectivity was discussed. The double bonds in 6 phenylhex-5-yn- 2-enoates were prone to migrate to the position conjugated with the carbon-carbon triple bonds to form 6-phenylhex-5-yn-3-enoates at higher temperature. The migration did not occur in absence of an excess amount of allenic/propargylic zinc reagent or at low temperature. PMID- 10789438 TI - On the radical brook rearrangement. Reactivity of alpha-silyl alcohols, alpha silyl alcohol nitrite esters, and beta-haloacylsilanes under radical-forming conditions AB - Two alkoxyl radical generation methods, lead tetraacetate treatment of alcohols and photolysis of nitrites, were applied to alpha-silyl alcohols 21 and to the corresponding nitrites 25 with a view to forming alpha-silyl alkoxyl radicals 23 and studying their possible radical Brook rearrangement to alpha-silyloxy carbon radicals 24. LTA treatment of 21 led to their quick and efficient conversion into mixed acetyl-silyl acetals 33 under very mild conditions. Photolysis of alpha alkylmonosubstituted alpha-silyl nitrites 25 to the corresponding aldehydes is considered to proceed through alpha-silyl alkoxyl radical intermediates 23. A concerted process is, however, proposed for the case of the benzyl nitrites analogues. On the basis of these results, it is postulated that resonance stabilization can have a major influence on the evolution of alpha-silyl alkoxyl radicals: should this stabilization be possible, they quickly evolve by alpha silyl fragmentation; otherwise, they tend to undergo radical Brook rearrangement. It was also found that the radical Brook rearrangement of alpha-silyl cyclopropyloxyl radicals generated from beta-bromoacylsilanes under standard tin radical conditions is too slow to compete with beta-fragmentation. PMID- 10789441 TI - Thermal decomposition reaction of acetone triperoxide in toluene solution AB - The thermal decomposition reaction of acetone cyclic triperoxide (3,3,6,6,9,9 hexamethyl-1,2,4,5,7,8-hexaoxacyclononane, ACTP) in the temperature range of 130.0-166.0 degrees C and an initial concentration of 0.021 M has been studied in toluene solution. The thermolysis follows first-order kinetic laws up to at least ca. 78% acetone triperoxide conversion. Under the experimental conditions, a radical-induced decomposition reaction as a competing mechanism may be dismissed, so the activation parameters correspond to the unimolecular thermal decomposition reaction of the ACTP molecule [delta H++ = 41.8 (+/- 1.6) kcal mol-1 and delta S++ = 18.5 (+/- 3.8) cal mol-1K-1]. Analysis of the reaction products are not enough to elucidate the real mechanism for the thermolysis of the acetone triperoxide in toluene solution. PMID- 10789440 TI - Solution-phase synthesis of a hindered N-methylated tetrapeptide using Bts protected amino acid chlorides: efficient coupling and methylation steps allow purification by extraction. AB - N-Benzothiazole-2-sulfonyl (Bts)-protected amino acid chlorides were used to prepare the hindered cyclosporin 8-11 tetrapeptide subunit 1. The synthesis was performed via 3a and the deprotected amines 5a, 13, and 19, including three repeated cycles involving N-methylation using iodomethane/potassium carbonate, deprotection of the Bts group, and N-acylation with a N-Bts-amino acid chloride such as 9b or 9c. Among three Bts cleavage methods compared (H3PO2/THF; NaBH4/EtOH; PhSH/K2CO3), the third gave somewhat higher overall yields. N Acylation of 5a with the Bts-protected N-methylamino acid chloride 10b followed by deprotection was also highly efficient and could be used as an alternative route to 11. Each of the deprotected amines was isolated without chromatography using simple extraction methods to remove neutral byproducts. The tetrapeptide 1 was obtained in analytically pure form as the monohydrate. PMID- 10789439 TI - Synthesis of C-C-bridged bis-isoflavones. AB - Unique C-C-bridged bis-isoflavones 5, 8, and 9 were obtained by reaction of 2 bromomethyl-7,4'-dimethoxyisoflavone 4 with ethyl cyanoacetate anion or tetraethylammonium cyanide or by Pd-catalyzed ethoxycarbonylation, respectively. The phenolic carboxylic acid 7 is available from 5 in two steps. PMID- 10789442 TI - Benzylic manganese halides, sulfonates, and phosphates: preparation, coupling reactions, and applications in organic synthesis AB - The use of highly active manganese, prepared by the Rieke method, for the direct preparation of benzylic manganese reagents was investigated. The oxidative addition of the highly active manganese (Mn) to benzylic halides was easily completed under mild conditions. More importantly, benzylic manganese sulfonates and phosphates were prepared by direct oxidative addition of Mn to the carbon oxygen bonds of benzylic sulfonates and phosphates. The resulting benzylic manganese reagents were found to undergo cross-coupling reactions with a variety of electrophiles. The majority of these reactions were carried out in the absence of any transition metal catalyst under mild conditions. This approach also provided a facile synthetic route to the preparation of resorcinolic lipids. PMID- 10789443 TI - Ab initio studies of AB - Ab initio calculations of the [1,5]-H shift in (3Z)-penta-1,3-diene and other substituted pentadienes and heteroanalogues using the hybrid density functional Becke3LYP with the 6-31G basis set are presented. Electron-donating substituents, such as methoxy in (3Z)-3-methoxypenta-1,3-diene 1, or heteroatoms such as a nitrogen atom in (Z)-ethylidenevinylamine 2, (1Z)-buta-1,3-dienylamine 3, (2Z) but-2-enylideneamine 4, (Z)-allylidenemethylamine 5, and methylene-(Z) propenylamine 6 are introduced. The electron-withdrawing fluoride is substituted for the hydrogen atoms in (3Z)-3-fluoropenta-1,3-diene 7, (3Z)-2,4-difluoropenta 1,3-diene 8, (3Z)-1,1',2,3,4,5,5'-heptafluoropenta- 1,3-diene 10, (1E,3E)-1,3,5 trifluoropenta-1,3-diene 11, and (1Z,3E)-1,3,5- trifluoropenta-1,3-diene 13. A detailed analysis of the geometries, energies, and electronic characteristics of the sigmatropic transposition compared to those of the unsubstituted case provides insights into substituent effects of this prototype of pericyclic reaction. The inductive and mesomeric effects of heteroatoms or heterosubstituents are of a great importance and in a continuous balance in the energetics of the transformation. Sterics can also play an important role due to the geometrical constraints of the reaction. As a general trend, decreasing the electron density of the phi system destabilizes the aromatic transition structure and increases the activation energy, and vice versa. PMID- 10789444 TI - Crystallization-induced asymmetric transformation of a tertiary phosphine AB - An equilibrating diastereomer mixture of the tertiary phosphines 5 and 6 (2.5:1 equilibrium ratio) undergoes crystallization-induced asymmetric transformation upon slow evaporation of solvent from refluxing heptane to give a 20:1 ratio in favor of the more stable crystalline isomer 5. The process can also be carried out at room temperature by using iodine to catalyze the interconversion of 5 and 6 via a pentavalent intermediate 8. However, this variation is more sensitive to the purity of the starting phosphine. Crystalline 5 can be converted to the stable borane complex 3, and reductive cleavage of the fluorenyl group using lithium naphthalenide affords the corresponding lithio derivative 10. Alkylation with iodomethane or benzyl bromide affords 13 or 17, respectively, with retention of phosphorus configuration. PMID- 10789445 TI - Copper(II)-catalyzed reactions of activated aromatics. AB - The catalytic reaction of cis-bisglycinato copper(II) monohydrate in the presence of hydrogen peroxide leads to hydroxylation of phenol to give catechol and hydroquinone (1:1.2 ratio) in good yield. 2,6-Dimethylphenol can be hydroxylated by hydrogen peroxide and a catalytic amount of cis-bisglycinato copper(II) monohydrate to give an aggregate of 1,4-dihydroxy-2,6-dimethylbenzene and 2,6 dimethylphenol. A similar reaction of o-cresol gives 2,5-dihydroxytoluene. The reactivity of cis-bisglycinato copper(II) monohydrate in hydrogen peroxide with o cresol is 4.5 times faster than that of a similar reaction by trans-bisglycinato copper(II) monohydrate. A catalytic reaction of cis-bisglycinato copper(II) monohydrate with aniline in aqueous hydrogen peroxide gives polyanilines in the form of pernigraniline with different amounts of Cu(OH)2 attached to them. The two major components of polyanilines obtained have Mn values of 1040 and 1500, respectively. Resistance of films of these polyanilines increases with temperatures from 40 degrees C to a maximum value at 103 degrees C and then decreases in the region of 103-150 degrees C, showing the property of a thermolectric switch. The aggregate prepared from hydroxylation of 2,6 dimethylphenol shows a similar property in the region of 30-180 degrees C. PMID- 10789446 TI - Determination of the photochemical efficiency of o-quinodimethane ring closure in room-temperature solutions by using time-delayed, two-color photolysis technique AB - Photochemical efficiency of o-quinodimethane (3) ring closure at room temperature was determined by using a time-delayed, two-color photolysis technique. o Quinodimethane (3) was generated by the photolysis of 1,2 bis[(phenylseleno)methyl]benzene (1) by a KrF (248 nm) laser pulse and thus generated 3 was photolyzed by a subsequent XeCl (308 nm)/XeF (351 nm) laser pulse with varying delay time of 0 to 3 s. The time profile of 3 was monitored by the chemical analyses of benzocyclobutene (5) (a photochemical product of 3), which was formed by a one-photon process, and the spiro dimer of 3 (4) (a thermal product of 3) in the two-color photolysis experiments. The time profile of 3 followed a second-order decay kinetics. The photochemical efficiency was obtained by the analysis of the delay-time dependence of the product yields; those of the consumption of 3 and the conversion 3-->5 by a single pulse of the excimer laser were 81% and 5.7% for the XeCl laser, and 73% and 2.3% for the XeF laser. This difference was attributed to the different excited states involved in the photolysis. In contrast to the photolysis of 3 in argon or rigid organic matrixes, it was revealed that photochemical conversion 3-->5 was not the main path in the solutions, and intermolecular reactions predominated. PMID- 10789447 TI - Fe(II)-catalyzed imidation of allyl sulfides and subsequent AB - Allyl aryl sulfides 1 and 5 were shown to undergo an imidation/[2,3]-sigmatropic rearrangement reaction upon treatment with N-tert-butyloxycarbonyl azide (BocN3) and catalytic amounts of FeCl2 in CH2Cl2. The N-Boc-protected N-allyl sulfenamides 3 and 21 were obtained in yields between 48 and 75% (12 examples). Whereas the reaction is well suited for the transformation of alpha-unbranched sulfides to alpha-branched sulfenamides, the enantiomerically pure alpha-branched sulfides 10 and 13 reacted sluggishly. The corresponding sulfenamides 22 and 23 were obtained in only moderate enantiomeric excess (36-39% ee). A reaction mechanism is proposed that postulates the intermediacy of an N-Boc-substituted Fe(IV)-nitrene complex 14 acting as the imidation reagent in the catalytic cycle. Possible side reactions are discussed. The benzenesulfenamides 3 were further converted into N-Boc-N-allylamines 4 by removal of the phenylsulfanyl group. Bu3SnH in benzene was found to be the reagent of choice for the deprotection of alpha-branched amines that bear a secondary allyl substituent (five examples, 71 93% yield). This method failed for the alpha-branched amines 3i-k with a tertiary allyl substituent. The phenylsulfanyl group was finally removed with P(OEt)3/NEt3 in CH2Cl2 (three examples, 43-62% yield). PMID- 10789449 TI - General synthetic entry to linearly-fused dihydrobenzocyclobutene-1,2-diones and benzocyclobutene-1,2-diones via annulation of 1,2-bis(methylene)carbocycles with 3-chloro-3-cyclobutene-1,2-dione AB - The tandem Diels-Alder/dehydrochlorination reaction of semisquaric chloride (1) with the 1,2-bis(methylene)cycloalkanes 2a-c and 1,2-bis(methylene)-4-cyclohexene (9) affords the linearly-fused cycloalkanodihydrobenzocyclobutene-1,2-diones 3a-c and 3,4,7,8-tetrahydrocyclobuta[b]-naphthalene-1,2-dione (10), respectively. On treatment with MnO2, 3a-c are dehydrogenated to the respective carbocycle-fused benzocyclobutene-1,2-diones 4a-c in good yields. 3a and 3b react with bromine to give the addition products 5a,b, which, on treatment with silver trifluoroacetate, afford the benzocyclobutene-1,2-diones 4a,b. For preparative purposes, the sequence 3-->5-->4 can be performed advantageously as a "one-pot procedure". Double-condensation reactions of 4a,b with alpha,alpha'-biscyano-o xylene and o-phenylenediamine afford the pentacyclic biphenylenes 7a,b and the cyclobutahetarenes 8a,b, respectively. These cyclobutenediones suggest themselves as building blocks for the construction of extended linearly-fused polycyclic compounds with novel ring sequences. o-Quinodimethanes 12a-g generated in situ by the thermal decomposition of the respective 1,4-dihydro-2,3-benzoxathiin-3-oxides (sultines) 11a-g react with semisquaric chloride (1) to afford the 3,8 dihydronaphtho[b]cyclobutene-1,2-diones 13a-g. These, on dehydrogenation with bromine and/or MnO2, furnish the naphtho[b]cyclobutene-1,2-diones 14a-g in fair to good yields. As described for 4a,b the naphtho[b]cyclobutene-1,2-diones 14a-c are condensed with alpha,alpha'-biscyano-o-xylene and o-phenylenediamine to furnish the pentacyclic biphenylenes 15a-c and the pentacyclic cyclobutahetarenes 16a-c. PMID- 10789448 TI - Using the Pummerer cyclization-deprotonation-cycloaddition cascade of imidosulfoxides for alkaloid synthesis. AB - The Pummerer reaction of imidosulfoxides bearing tethered alkenyl groups has been employed for the synthesis of several alkaloids. The required imidosulfoxides necessary for the cascade sequence were easily obtained by heating the appropriate amide with (ethylsulfeny)acetyl chloride followed by sodium periodate oxidation. The initially formed thionium ion, obtained by treating the imidosulfoxide with acetic anhydride and p-toluenesulfonic acid, reacts with the neighboring imido group, and the resulting oxonium ion undergoes subsequent deprotonation to produce an isomunchnone dipole. This mesoionic betaine intermediate undergoes ready intramolecular dipolar cycloaddition across the neighboring pi-bond. Exposure of the resulting cycloadducts to additional acetic anhydride leads to ring opening and formation of a 5-acetoxy-substituted 2(1H) pyridone. This six-ring heterocyclic system constitutes a valuable building block for the synthesis of a variety of pyridine, quinolizidine, and clavine alkaloids. The cyclization-deprotonation-cycloaddition cascade has been successfully applied to the synthesis of the naturally occurring alkaloids onychnine, dielsiquinone, (+/-)-lupinine, (+/-)-anagyrine, (+/-)-pumiliotoxin C, and (+/-)-costaclavine. PMID- 10789450 TI - Synthesis and metal ion complexation studies of proton-ionizable calix AB - A series of novel N-chromogenic calix[4]arene azacrown ethers were synthesized as selective extractants of potassium ion. 1,3-Alternate calix[4]arene azacrown ethers were prepared by reacting 25,27-dipropyloxy-26,28-bis(5-chloro-3 oxapentyloxy) calix[4]arenes with p-toluenesulfonamide in the presence of potassium carbonate. The coupling reaction of calix[4]arene azacrown ether with 2 hydroxy-5-nitrobenzyl bromide in the presence of triethylamine in THF gave the chromogenic calix[4]arene azacrown ether in moderate yield. These compounds show high potassium selectivity over other metal ions as shown by two-phase extraction, bulk liquid membrane, and 1H NMR studies on a ligand-metal complex. It is assumed that the OH of the chromogenic group attached on nitrogen can assist the complexation by encapsulation of the metal. PMID- 10789451 TI - Synthesis of 1,1-linked galactosyl mannosides carrying a thiazine ring as mimetics of sialyl Lewis X antigen: investigation of the effect of carboxyl group orientation on P-selectin inhibition. AB - This paper describes the synthesis of 1,1-linked galactosyl mannosides as sialyl Lewis X mimetics that contain a spiro-ring to position the carboxylate group in a well-defined orientation. It was found that compound 4 is more active as a P selectin inhibitor (IC50 = 19 microM) than the parent disaccharide 2, which contains a flexible carboxyl group (IC50 = 193 microM). This result is consistent with that observed in the previous NMR study of sialyl Lewis X bound to P selectin. The chemistry described here should be useful for the development of selective inhibitors of E-, P-, and L-selectins. PMID- 10789452 TI - Synthesis of 2-substituted polyhydroxytetrahydropyrimidines (N-hydroxy cyclic guanidino-sugars): transition-state mimics of enzymatic glycosidic cleavage. AB - The synthesis of 2'-substituted polyhydroxytetrahydropyrimidines as transition state mimics of enzymatic glycosidic cleavage has been achieved by using guanylation and cyclization methodologies. The D-galacto type N-hydroxy cyclic guanidino-sugar 21 was synthesized in six steps from amine 7 and thiourea 14 in an overall yield of 59%. To further derivatize compound 21 to incorporate the leaving group moiety, we have synthesized 2-methylsulfanyl compounds 26-29 as key intermediates. The 2-methylsulfanyl group in 29 was displaced with amines, assisted by silver tetrafluoroborate as Lewis acid, to give protected cyclic guanidines 30-32 in moderate yields (60-67%). Removal of the protecting groups in 32 gave the D-galacto-type N-hydroxy cyclic guanidino-sugar 34. The key steps in the synthesis of the 6-deoxy-DL-galacto type N-hydroxy cyclic guanidino-sugars 49, 54, and 64-66 involve cyclization of the appropriate acetal intermediates (45, 50, and 58-60) followed by removal of the protecting groups. PMID- 10789453 TI - Anomeric reactivity-based one-pot oligosaccharide synthesis: a rapid route to oligosaccharide libraries. AB - The assembly of an oligosaccharide library has been achieved in a practical and efficient manner employing a' one-pot sequential approach. With the help of the anomeric reactivity values of thioglycosides, using a thioglycoside (mono- or disaccharide) with one free hydroxyl group as acceptor and donor coupled with another fully protected thioglycoside, a di- or trisaccharide is selectively formed without self-condensation and subsequently reacted in situ with an anomerically inactive glycoside (mono- or disaccharide) to form a tri- or tetrasaccharide in high overall yield. The approach enables the rapid assembly of 33 linear or branched fully protected oligosaccharides using designed building blocks. These fully protected oligosaccharides have been partially or completely deprotected to create 29 more structures to further increase the diversity of the library. PMID- 10789454 TI - Self-folding molecules: a well defined, stable loop formed by a carboxylate guanidinium zwitterion in DMSO AB - Two zwitterions 1a,b have been synthesized, in which a carboxylate group is attached via a flexible alkyl chain of different length (butylene and ethylene, respectively) to a guanidiniocarbonyl pyrrole cation moiety. For 1b, no signs for either an intra- or intermolecular association between these two groups in polar solution (DMSO) could be found. In contrast to this, the 1H NMR spectrum of 1a shows clear evidence for a strong interaction between the carboxylate and the guanidiniocarbonyl pyrrole cation. According to variable-temperature and concentration-dependent concentration-dependent NMR studies, this interaction stems from an intramolecular complexation. It was shown by ROESY and H/D-solvent exchange experiments that 1a, even in DMSO, folds into a well-defined intramolecular loop conformation held together by multiple weak interactions. PMID- 10789455 TI - A highly stereoselective synthesis of 11Z-retinal using tricarbonyliron complex. AB - A stereoselective synthesis of 11Z-retinal 2, which is the chromophore of visual pigment (rhodopsin), was accomplished from the beta-ionylideneacetaldehyde tricarbonyliron complex 3. The Peterson reaction of 3 using ethyl trimethylsilylacetate smoothly proceeded to afford predominantly the Z-ester 6. Transformation of the Z-ester 6 to the methyl ketone 19, followed by the Emmons Horner reaction of 19 with C2-cyanophosphonate, produced ethyl 11Z, 13E retinonitrile-tricarbonyliron complex 21 as the only product. Decomplexation of 21 with CuCl2 and subsequent DIBAL reduction gave 11Z-retinal 2 in excellent yield. Mechanistic consideration of Z-selectivity in the Peterson reaction of the aldehyde-tricarbonyliron complex is also discussed. PMID- 10789456 TI - Lewis acid-directed cyclocondensation of piperidone enol ethers with 2-methoxy-4 (N-phenylsulfonyl)-1,4-benzoquinoneimine: a new regioselective synthesis of oxygenated carbolines AB - Lewis acid-directed cyclocondensations of piperidone enol ethers with 2-methoxy-4 (N-phenylsulfonyl)-1,4-benzoquinoneimine are reported. Benzofurans are obtained with BF3.OEt2 as a promoter, whereas use of excess amounts of TiCl4:Ti(OiPr)4 leads to tetrahydrocarbolines. The latter reactions provide expedient routes to oxygen-substituted tetrahydrocarbolines and carbolines. As applications of this new methodology, the preparations of 1-[3-(dimethylamino)propyl]amino-7-methoxy- and 1-[3-(dimethylamino)propyl]amino-7,8-dimethoxy-5H-pyrido[4,3-b]indoles are described. PMID- 10789457 TI - Tandem reduction-chloroallylboration of esters: asymmetric synthesis of lamoxirene, the spermatozoid releasing and attracting pheromone of the laminariales (Phaeophyceae). AB - The asymmetric synthesis of all four stereoisomers of lamoxirene (cis-2 cyclohepta-2,5-dienyl-3-vinyloxirane), the spermatozoid-releasing and -attracting pheromone of the Laminariales (Phaeophyceae), is reported. Chiral ethyl cyclohepta-2,5-diene carboxylates, prepared by a divinylcyclopropane Cope rearrangement, were effectively alkylated by means of a novel tandem DIBAL-H reduction/asymmetric alpha-chloroallylboration using (Z)-gamma chloroallyldiisopinocampheylboranes. The ensuing syn-alpha-chlorohydrins were transformed into the corresponding vinyloxiranes with DBU, providing all four isomers of the pheromone in good chemical and excellent optical yield (90-97% ee). Spermatozoid-release assays were conducted with the sympatrically growing species L. digitata, L. hyperborea, and L. saccharina and established (1'S,2R,3S) 1c as the most active isomer in all cases. PMID- 10789458 TI - Theoretical study of activation of oxirane by bidentate acids AB - The reaction of oxirane with amines in the presence of some bidentate and monodentate Bronsted acids has been studied theoretically by taking some simplified reaction models. Acids have been shown to serve as temporary reservoirs of electronic charge during the reaction to strengthen the nucleophilicity of the substrate. The shift of electronic charges and enhancement of the nucleophilicity of oxirane have been represented by means of paired interaction orbitals and projected reactive orbitals. The reason the transition state appears at an earlier stage in the reaction catalyzed by 1,8 biphenylenediol relative to the reactions catalyzed by monodentate phenols and alcohols has been clarified. PMID- 10789459 TI - Bis(dimethylamino)porphyrazines: synthetic, structural, and spectroscopic investigations. AB - The synthesis and isolation of unsymmetrical porphyrazines bearing two, four, and six bis-(dimethylamino) functionalities has been achieved via the base-catalyzed cross-condensation of 1,2-dicyanobenzene 8 and bis(dimethylamino)maleonitrile 7. In addition, the benzo-fused hexaaminoporphyrazine dimer 10 was prepared from condensation of dinitrile 7 (in excess) with benzenebis(1,3-diiminopyrroline) 9. Electrochemical studies reveal that all porphyrazines may be readily oxidized. The X-ray structures of porphyrazines 2b and 5a and the cis isomer 3a are presented. The latter is the first structure of a porphyrazine having a cis-type substitution pattern. The extended pi-conjugation in dimer 10 causes a approximately 100 nm red-shifted Q-band in the electronic absorption spectrum. PMID- 10789460 TI - Total synthesis of ningalin B utilizing a heterocyclic azadiene Diels-Alder reaction and discovery of a new class of potent multidrug resistant (MDR) reversal agents. AB - A concise, efficient approach to the total synthesis of ningalin B (1) based on a heterocyclic azadiene Diels-Alder strategy (1,2,4,5-tetrazine-->1,2,-diazine- >pyrrole) ideally suited for construction of the densely functionalized pyrrole core found in the natural product is detailed. Examination of the natural product and a number of synthetic intermediates revealed that while lacking inherent cytotoxic activity, many reverse the multidrug-resistant (MDR) phenotype, resensitizing a human colon cancer cell line (HCT116/VM46) to vinblastine and doxorubicin at lower doses than the prototypical agent verapamil. PMID- 10789461 TI - Conformationally constrained substance P analogues: the total synthesis of a constrained peptidomimetic for the Phe7-Phe8 region. AB - A lactam-based peptidomimetic for the Phe7-Phe8 region of substance P has been synthesized. The synthesis used an anodic amide oxidation to selectively functionalize the C5-position of a 3-phenylproline derivative. The resulting proline derivative was coupled to a Cbz-protected phenylalanine, and an intramolecular reductive amination strategy used to convert the coupled material into a bicyclic piperazinone ring skeleton. The net result was a dipeptide building block that imbedded one of two proposed receptor bound conformations for the Phe7-Phe8 region of substance P into a bicyclic ring skeleton. The building block was then converted into a constrained substance P analogue with the use of solid-phase peptide synthesis. A similar intramolecular reductive amination strategy was used to synthesize a substance P analogue having only Phe7 constrained, and the original 3-phenylproline was converted into a substance P analogue having only Phe8 constrained. All of the analogues were examined for their ability to displace substance P from its NK-1 receptor. PMID- 10789462 TI - Ab initio study on the photochemical isomerization of furan derivatives AB - The photochemical isomerization reactions of furan, 2-methylfuran, 2 trimethylsilylfuran, and furan-2-carbonitrile were studied using ab initio methods. The results are in agreement with the previously reported data obtained through semiempirical methods. In particular, the sensitized irradiation of furan derivatives populates the first excited triplet state of the furan, and this triplet state can evolve only through O-Ca cleavage. The selection of the bond to be broken can depend on energetic factors (furan-2-carbonitrile) or on kinetic factors (2-methylfuran, 2-trimethylsilylfuran). The direct irradiation of furan derivatives populates the singlet excited state and leads to a conversion into the Dewar isomer or into the corresponding triplet state through the usual intersystem crossing procedure. The efficiency of these processes determines the presence or the absence of isomerized furan derivatives in the reaction mixtures. PMID- 10789463 TI - Modification of regioselectivity in cycloadditions to C70 under microwave irradiation AB - The regioselectivity of the cycloaddition of N-methylazomethine ylide to C70 can be modified by using microwave irradiation as the source of energy. Under microwave irradiation and by choosing the appropriate solvent and irradiation power, the 5-6 isomer is the major product, a situation that is in contrast to conventional heating where the 1-2 isomer predominates. Moreover, isomer 7-21, which represents 13% of monoadducts under classical heating, is not formed under microwave irradiation and with ODCB as solvent. Theoretical calculations predict an asynchronous mechanism and suggest that the modification of the regiochemical outcome is related to the relative energies and hardnesses of the transition structures involved. PMID- 10789464 TI - Photochemistry of 4'-benzophenone-substituted nucleoside derivatives as models for ribonucleotide reductases: competing generation of 3'-radicals and photoenols. AB - Ribonucleotide reductases (RNRs) catalyze the 2'-reduction of ribonucleotides, thus providing 2'-deoxyribonucleotides, the monomers for DNA-biosynthesis. The current mechanistic hypothesis for the catalysis effected by this class of enzymes involves a sequence of radical reactions. A 3'-hydrogen abstraction, effected by a radical at the enzyme's active site, is believed to initiate the catalytic cycle. As models for this substrate-enzyme interaction, the photochemically induced intramolecular hydrogen abstraction in a series of 4' benzophenone-substituted nucleoside analogues was studied. Model compounds with hydroxy-, methoxy-, mesyloxy-groups or a cyclic carbonate in 2'- and 3'-positions were investigated. Depending on the substitution pattern, two different types of photoproducts were observed: Those which result from photoenol formation (gamma-H abstraction) and those which result from abstraction of the 3'-H-atom (delta-H abstraction). Photoenol formation was further supported by H/D-exchange experiments. Thus, the 3'-H-abstraction postulated as the initial step in RNR action was successfully modeled by photolysis of 4'-benzophenone-substituted nucleoside analogues. The regioselectivity of the photochemical H-abstraction and thus of the product distribution as a function of the 2'- and 3'-substituents was rationalized on the basis of a conformational analysis of the four model systems, utilizing molecular mechanics simulations. PMID- 10789465 TI - Nondynamic kinetic resolution of configurationally stable biaryl lactones by reduction with oxazaborolidine-activated borane: AM1 studies and experimental verification AB - The complete mechanistic course of the atroposelective ring opening of a lactone bridged biaryl, dinaphth[2,1-c:1',2'-e]oxepin-3-(5H)-one (3), with a chiral oxazaborolidine-BH3 complex was calculated using the semiempirical AM1 method. The first hydride transfer to the activated carbonyl function of the adduct complexes was elaborated to be the selectivity-determining step in the postulated five-step mechanism. The calculated enantioselectivity is in good accordance with the experimental results, so that related calculations were performed on the atroposelective ring opening of a sterically strongly hindered and therefore also configurationally stable six-membered biaryl lactone, 1,3-di-tert-butyl-6H benzo[b]naphtho[1,2-d]pyran-6-one (6f). These calculations predicted a highly (M) selective reduction of 6f (kM/kP = 358 at -78 degrees C), which, after the smooth preparation of 6f by intramolecular biaryl coupling in high yields, was fully confirmed experimentally (kM/kP > 200 at -78 degrees C). Isolation of the intermediate hydroxy aldehyde (M)-14 at the beginning of the reaction with the same enantiomeric excess as found for the corresponding alcohol (M)-7f conclusively showed the first hydride transfer step to determine the selectivity of this process. The good agreement of computationally predicted and experimentally confirmed values proves the suitability of the AM1 method for mechanistic studies on even such complex reactions and opens a most efficient overall synthesis of sterically highly hindered biaryls, in excellent chemical (for the ring closure) and optical (for the ring cleavage) yields and for any desired axial configuration. PMID- 10789466 TI - Unmasking the 6,7,5-tricarbocyclic frame of AB - Opening of the bridging ether of 8-oxabicyclo[3.2.1]oct-6-ene systems 1,7-fused to a five-membered carbocycle that bears a double bond conjugated to that of the oxabicycle can be induced at low temperaure by the addition of an organolithium reagent. If this double bond is unsubstituted, the opening occurs via an anti-1,6 addition pathway, and if substituted it takes place by means of a more classical syn SN2' addition. PMID- 10789467 TI - Asymmetric copper-catalyzed AB - The asymmetric copper-catalyzed generation and subsequent [2,3]-sigmatropic rearrangement of sulfur ylides is strongly dependent on the structure of the starting allyl sulfide. A series of alkyl and aryl substituted allyl sulfides (2a i) were reacted with ethyl diazoacetate in the presence of copper triflate (CuOTf) and a C2-symmetric bis-oxazoline ligand (5a-c). The degree of asymmetric induction ranged from 2.8% for allyl methyl sulfide (2a) to 60% for (1S,2S,5R) (+)-allyl menthyl sulfide (2d). The enantioselectivity of the reactions was also dependent on the electronic nature of the sulfide; allyl phenyl sulfide (2e) gave a 14% ee, whereas allyl p-methoxyphenyl sulfide (2i) produced only an 8% ee. The stereochemistry of 2d and (1R,2S,5R)-(+)-allyl menthyl sulfide (7) was assigned on the basis of NMR spectroscopic experiments. PMID- 10789468 TI - Reactions within association complexes: the reaction of imidazole with substituted phenyl acetates in the presence of detergents in aqueous solution AB - The bimolecular rate constants for reaction of imidazole with phenyl acetates complexed with sodium dodecyl sulfate (SDS) or cetyltrimethylammonium bromide (CTAB) micelles obey Bronsted equations with beta 1g similar to that of the reaction in aqueous solution. The dissociation constants of ester (Ks) and the hypothetical dissociation constant (KTS) of the transition state of the micelle complexes obey Hansch equations with similar sensitivities (p) to pi (-0.66 and 0.589 for KS and -0.735 and -0.495 for KTS, respectively). The slopes also indicate that the microsolvation environments associated with the transition state and the complexed ester have aqueous character. The relative values of KTS and KS indicate that the transition state of the reaction of imidazole with ester is more weakly complexed to both micelles than is the reactant ester. Log KTS values are linear functions of log KS for reactions with both CTAB and SDS; the slopes are, respectively, -0.893 and -1.19 consistent with a slightly more "water like" medium for the transition state than for the site of binding of ester with CTAB-micelle and slightly less for the SDS-micelle. The results for ester and transition state are consistent with the location of the phenyl residue in a hydrophobic region that possesses water molecules. It is concluded that the acetyl group in the complexed transition state is located in an aqueous part of the Stern region, whereas the phenyl residue is in a part of the Stern region that possesses alkane components. The derived kinetic and complexation parameters in these experiments refer to micelles with Stern regions that have been maintained at constant ionic compositions. PMID- 10789470 TI - Correlation of the rates of solvolysis of cyclopropylcarbinyl and cyclobutyl bromides using the extended Grunwald-Winstein equation AB - The reactions of cyclopropylcarbinyl bromide (1) and cyclobutyl bromide (2) in hydroxylic solvents proceed with both solvolysis and rearrangement. Depending on the solvent, the reactions of 1 are 10-120 times faster than those of 2, and both are faster than the previously studied allylcarbinyl bromide (3). Specific rates are reported for the reactions of 2 proceeding to solvolysis products and 3. Reactions of 1 proceed to solvolysis products and both 2 and 3; since 2 slowly undergoes further solvolysis, specific rates are obtained by a modified Guggenheim treatment. The two sets of specific rates are analyzed using the extended Grunwald-Winstein equation to give sensitivities toward changes in solvent nucleophilicity of 0.42 for 1 and 0.53 for 2 and corresponding sensitivities toward changes in solvent ionizing power of 0.75 and 0.94. A mechanism is proposed involving a rate-determining ionization with an appreciable nucleophilic solvation of the incipient carbocation. PMID- 10789469 TI - Synthesis of the C-glycoside analogue of a novel sialyl Lewis X mimetic. AB - Sialyl Lewis X (sLex) mimetics that can function as selectin antagonists have received considerable attention in connection with the development of novel antiinflammatory therapies. An interesting structure that emerged from the studies of the Wong group is the 1,1-Gal-Man disaccharide 2, reported to bind E selectin 5 times more strongly than sLex. The C-glycoside derivative 3 is of interest both as a conformational probe for selectin binding and as a hydrolytically stable analogue. Herein we illustrate a novel methodology for beta C-galacto-disaccharides in the synthesis of 3. The protocol has as a key step a novel oxocarbenium ion-enol ether cyclization to give a C1-substituted galactal. PMID- 10789471 TI - Vinyl benzenes as dienes in mild solid-phase Diels-Alder reactions AB - Wang resin-bound intermediates derived from Fmoc-L-phenylalaninal and Fmoc-L valinal, and a resin supported Horner-Wadsworth-Emmons reaction, were treated with cinnamaldehyde derivatives, acetic acid, and borohydride to give secondary amines which were subsequently benzoylated to afford various derivatives of 3. Heating 3 at 95 degrees C induced cycloaddition reactions and produced 4 as the major product. Compounds 3 which were derived from 4-methoxycinnamaldehyde were more reactive, but did not give 4 and 4-7g. The direct cleavage of 3b using TFA led to the isolation of cycloaddition-demethylation product 10. The derivative of 3, which contained an electron-withdrawing nitro group on the phenyl ring, produced a single diastereomer of 4. The Diels-Alder cycloaddition between two electron-deficient counterparts showed similar reactivity to that of the reactions which have a normal complementary electron-demand. PMID- 10789473 TI - beta-scission of tertiary alkyl hypochlorites promoted by phase-transfer catalysis PMID- 10789472 TI - Synthesis of phytyl- and chroman-derivatized photoaffinity labels based on alpha tocopherol. AB - Photoaffinity analogues of alpha-tocopherol have been prepared by substituting photosensitive functional groups at either the terminus of an alkyl chain of varying length mimicking the phytyl tail or on C-3 of the chroman portion of tocopherol. The alkyl chain-modified compounds 2a-d contain a hexyl to nonyl alkyl chain extending from C-2 of the chroman, terminating in a tetrafluoroazidobenzyloxy group. These compounds were prepared starting from the commercially available Trolox acid 4, followed by esterification, protection, and reduction to the silyl-protected Trolox aldehyde 7, which was coupled using Wittig chemistry to different omega-hydroxyphosphonium bromides. Reduction of the alkene product, coupling with p-azidotetrafluorobenzyl bromide, and deprotection of the phenolic silyl group gave compounds 2a-d in excellent yields. Chroman functionalized photoaffinity labels were synthesized starting from the protected tocopherol chromene 16b which was a key intermediate for preparation of a 3 hydroxy derivative, either by reduction of epoxides produced directly with Jacobsen's catalysts or by treatment with NBS in wet DME to give two stereoisomeric bromohydrins which were cyclized and reduced to give the phenol protected C-3 alcohols 19a,b. These alcohols were then converted to diazoacetate esters, and the protecting group was removed to give 3-diazoacetoxy alpha tocopherols 3a,b. PMID- 10789474 TI - An easy access to 2,6-dihydroxy-9-azabicyclo PMID- 10789475 TI - An improved synthesis of selectively protected L-dopa derivatives from L tyrosine. PMID- 10789476 TI - Furan-fused TCNQ and DCNQI: synthesis and properties PMID- 10789477 TI - Unexpected one-pot epoxy sulfone-enaminone transformation. Synthesis of 5a-carba beta-mannopyranosylamine PMID- 10789478 TI - Efficient large-scale radical deoxygenation in alcoholic solvents using sodium hypophosphite and a phase-transfer agent: application to erythromycins. PMID- 10789479 TI - Asymmetric synthesis of beta-hydroxy esters and alpha-alkyl-beta-hydroxy esters by recombinant Escherichia coli expressing enzymes from baker's yeast. PMID- 10789480 TI - Practical synthesis of 1,6-anhydro-2,4-dideoxy-beta-D-glycero-hexopyranos-3-ulose from levoglucosan. PMID- 10789481 TI - Improved methodology for the generation and trapping of alpha-lactams by weak nucleophiles PMID- 10789482 TI - Conformational studies by dynamic NMR. 72. Stereolabile enantiomers of acyl and thioacyl ferrocenes PMID- 10789483 TI - Highly selective synthesis of 1-N-iminosugars of the D-glucose and -glucuronic acid types. PMID- 10789484 TI - A short, facile synthesis of 5-substituted 3-amino-1H-pyrrole-2-carboxylates. PMID- 10789485 TI - A short route to (-)-mintlactone by thallium(III)-mediated cyclization of (-) isopulegol. PMID- 10789486 TI - Ring-closing alkyne metathesis. Stereoselective synthesis of the cytotoxic marine alkaloid motuporamine C. PMID- 10789487 TI - beta-hydrogen-containing sodium alkoxides as suitable bases in palladium catalyzed aminations of aryl halides PMID- 10789488 TI - An efficient protocol for the preparation of primary alcohols bearing a beta chiral center via an oxazolidinone auxiliary mediated resolution, and application to the synthesis of 4,4-disubstituted piperidine substance P antagonists. PMID- 10789489 TI - Cancer chemoprevention. PMID- 10789490 TI - Chemoprevention: general perspective. AB - Chemoprevention is the use of natural or synthetic compounds to block, reverse, or prevent the development of invasive cancers. Cellular carcinogenesis forms the biologic basis for the identification of chemopreventives, assessment of their activity, and ultimately the success or failure of a chemopreventive. Chemopreventive agents undergo multistep evaluations to assess efficacy that are similar in concept but vastly different in practice to standard ablative oncologic therapeutics. In vitro assessments of potential anticarcinogenesis efficacy include measurements of an agent's antioxidant activity, induction of phase II metabolizing enzymes and effects upon cellular proliferation and apoptotic control pathways. In vivo efficacy is assessed primarily in rodent models of carcinogenesis that are specific for a given organ target. The role of genetically modified animal models in the in vivo assessment of chemoprevention agents remains unclear. Clinical assessment of chemopreventive agent efficacy consists of a multistep process of identification of an optimal chemopreventive agent (phase 1), demonstration of efficacy in humans through the modulation of reversal of a tissue, biochemical, and molecular surrogates for neoplastic transformation and invasion (phase 2) and cancer risk reduction in large cohort trials (phase 3). Opportunities and future needs include the development of reliable, predictive in vivo models of carcinogenesis, careful exploration of the preventive pharmacology of therapeutic agents being used for non-cancer prevention indications, and the incorporation of genetic risk cohorts to define cancer chemopreventive efficacy. PMID- 10789491 TI - Prevention of cervix cancer. AB - Cervix carcinoma is an important health problem world-wide, being the second most common cancer among women, ranking first in many developing countries. A number of important epidemiological risk factors have been identified as contributing to the development of CIN and invasive cervix carcinoma. Of key importance is infection with human papillomavirus (HPV), which is the primary risk factor. There are evolving primary and secondary preventive strategies that could further reduce the burden from cervical carcinoma. The possible primary preventive strategies include risk reduction, diet or dietary supplements, HPV vaccines, and other chemopreventive agents. The possible advances in secondary preventive strategies include new technologies for Pap smears, HPV typing triage, and other adjuvant screening procedures. The impact of these strategies will depend upon evidence to support their use along with the characteristics of the population and environment in which they are used. PMID- 10789492 TI - Prevention of lung cancer. AB - Lung cancer is the major cause of death in industrialized western societies. Its link to tobacco abuse is well established and efforts should be made to eliminate this potent environmental carcinogen. The concept of chemoprevention, the use of agents to inhibit and reverse lung cancer carcinogenesis, has great appeal. The CARET study, conducted in 18,000 high-risk smokers in the US, found that a combination of beta-carotene and retinyl palmitate resulted in a 28% increase in the incidence of lung cancer. A similar study conducted in Finland, the ATBC trial utilizing alpha tocopherol and beta-carotene, had similar findings for the group taking beta-carotene. These two trials have caused a rethinking of the use of natural compounds as chemoprevention agents. These agents should no longer be regarded as harmless, but as having potential toxicities. A new approach in the chemoprevention of cancer has been the concept of surrogate endpoints, biological changes that are on the pathway to cancer. Trials are underway to determine what are appropriate surrogate endpoints for lung cancer chemoprevention trials. PMID- 10789493 TI - Chemoprevention for colorectal cancer. PMID- 10789494 TI - Prevention of breast cancer. AB - With the advent of screening and the increased incidence of breast cancer, concern for the prevention of breast cancer has become forefront in today's society. Determining individual risk is the key to prescribing prevention. Prevention of breast cancer is still under clinical investigation with only one drug, tamoxifen, showing benefit in high risk patients. This paper reviews the possible sites for prevention of neoplastic transformation via biomarkers in a breast cell as well as the investigational drugs and their potential use in the chemoprevention of breast cancer. PMID- 10789495 TI - Expression and regulation of cytochrome P450 enzymes in primary cultures of human hepatocytes. AB - The aim of this study was to test suitable culture conditions for maintaining normal cellular cytoarchitecture and inducibility of P450 enzymes in primary cultures of human hepatocytes by prototypical inducers. The selectivity and sensitivity of a sandwich culture system were determined by treating cultures with a number of clinically relevant drugs that are known to be inducers of either rodent and/or human P450 enzymes. The results showed that considerable induction of CYP3A4 activity is observed at DMSO concentrations greater than 0.1% (v/v). No differences in P450 induction response were observed between cultures maintained under different matrix conditions. However, the matrix condition considered to be optimal for maintaining cellular integrity, protein yields, and P450 enzyme induction was a sandwich configuration in combination with modified Chee's medium containing insulin (6.25 microg/mL) and dexamethasone (< or =0.1 microM). Under these conditions, induction of CYP3A4 occurred at clinically relevant drug concentrations, and maximal activities were achieved after 3 days of exposure. Overall, the response of human hepatocyte cultures to treatment with both positive and negative modulators was found to reflect that observed in vivo with respect to both enzyme specificity and potency of enzyme induction, although considerable sample-to-sample variability was observed in the magnitude of induction. PMID- 10789496 TI - Effects of new antioxidant compounds PNU-104067F and PNU-74389G on antioxidant defense in normal and diabetic rats. AB - Diabetes mellitus and its complications are associated with elevated oxidative stress, leading to much interest in antioxidant compounds as possible therapeutic agents. Two new classes of antioxidant compounds, the pyrrolopyrimidines and the 21-aminosteroids, are known to inhibit lipid peroxidation and other biomolecular oxidation. We hypothesized that in the presence of excess oxidants or the impaired antioxidant defense seen in diabetes mellitus, administration of antioxidants such as these may reverse the effects of diabetes on antioxidant parameters. This study measured the effects of subchronic (14 day) treatment with a pyrrolopyrimidine (PNU-104067F) or a 21-aminosteroid (PNU-74389G) in normal and diabetic Sprague-Dawley rats. Activity levels of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, concentrations of oxidized and reduced glutathione, and lipid peroxidation were used as measures of antioxidant defense in liver, kidney, heart, and brain tissue. In normal rats, the only effect was a 43% increase in cardiac lipid peroxidation after treatment with PNU 104067F. In diabetic rats, the only reversals of the effects of diabetes were a 30% decrease in hepatic glutathione peroxidase activity after PNU-74389G treatment and a 33% increase in cardiac glutathione disulfide concentration after PNU-104067F treatment. In contrast to these effects, increased cardiac glutathione peroxidase and catalase activities, increased brain glutathione peroxidase activity, increased hepatic lipid peroxidation, decreased hepatic glutathione content, and decreased hepatic catalase activity were seen in diabetic rats, reflecting an exacerbation of the effects of diabetes. PMID- 10789497 TI - Identification of a putative ribosomal protein mRNA in Chironomus riparius and its response to cadmium, heat shock, and actinomycin D. AB - A putative ribosomal protein (rp) mRNA in Chironomus riparius has been found using differential display (DD). Its sequence has 84.8% identity with mosquito rp L8, Aedes albopictus, and is approximately 0.9 kb. Studies were undertaken in order to evaluate rp as a control for environmentally relevant genes. Responses of Drosophila heat shock 70 gene (hsp70) were used to establish heat shock temperatures and cadmium (Cd) concentrations for Chironomus experiments and to validate DD. Expression of hsp70 was induced over control by 28 degrees C at 30 minutes and 1 mM Cd at 24 hours (p< or =0.05). For Chironomus, DD, Northern blot, and nuclease sensitivity were used to measure responses to two stressors: heat shock for 30 minutes and Cd for 24 or 48 hours. Differential display and nuclease sensitivity assays found expression of rp mRNA at 37 degrees C and 16 mM Cd to be similar to controls. Northern blots indicated statistically significant effects for heat shock (p = 0.046) but not Cd (p = 0.406). However, mRNA levels at 37 degrees C were increased only 1.72-fold over controls. A concentration of 24 nM actinomycin D suppressed rp expression as measured by nuclease sensitivity assays. Stressors should not affect rp mRNA levels below their LC-50s. PMID- 10789499 TI - Biliary copper excretion capacity in intact animals: correlation between ATP7B function, hepatic mass, and biliary copper excretion. AB - Copper toxicosis can occur in the absence of biliary copper excretion. To demonstrate whether biliary copper excretion capacity is correlated with hepatic mass and ATP7B function, we undertook studies in intact animals. Copper-histidine was injected intrasplenically after baseline bile collection, followed by measurement of copper excretion in Long-Evans Cinnamon (LEC) rats lacking atp7b function and in normal, syngeneic Long-Evans Agouti (LEA) rats. The basal biliary copper excretion was very low in LEC rats compared with LEA rats, 8+/-4 and 37+/ 18 ng copper/min, respectively; p<0.05. After addition of copper, copper excretion increased significantly (by two- to five-fold) in LEA rats during the 30 minute study period, whereas LEC rats showed only a slight and transient increase in copper excretion. After one-third and two-thirds partial hepatectomy immediately before copper loading, copper excretion decreased progressively. The studies indicate that biliary copper excretion depends on hepatocyte mass and ATP7B gene function. Analysis of copper excretion with our non-radioactive method will facilitate testing of novel therapies and pathophysiological mechanisms in copper toxicity. PMID- 10789498 TI - Induction of CYP1A by carbofuran in primary culture of fish hepatocytes. AB - Carbofuran is a nematicide used in agricultural fields throughout the world. Indiscriminate use of this pesticide poses severe detrimental effects on our ecosystem. We have shown that it induces the CYP1A (cytochrome P4501A) monooxygenase enzyme system in cultured hepatocytes from Indian catfish, Heteropneustes fossilis (Bloch). We have quantified this induction by measuring the activity of the enzyme 7-ethoxyresorufin-O-deethylase (EROD), synthesized from CYP1A1 gene. The induction followed a dose-dependent relationship with carbofuran. The dose-dependent curve of EROD using carbofuran was very much similar with beta-napthoflavone, which is a known inducer of CYP1A1. Coexposure of these compounds to the culture media showed a synergistic effect on the enzyme activity. A blocker of aromatic hydrocarbon receptor, alpha-napthoflavone, blocked carbofuran-induced EROD activity in a dose-dependent manner. All these findings suggest that metabolism of carbofuran might be mediated by the CYP1A monooxygenase system through binding of the aromatic hydrocarbon receptor. We have also studied the superinduction phenomenon, which is a typical characteristic of the CYP1A gene in our system. PMID- 10789500 TI - Carnosine prevents the glycation-induced changes in electrophoretic mobility of aspartate aminotransferase. AB - Carbohydrate-derived aldehydes cause irreversible loss of protein function via glycation. We previously observed that glyceraldehyde 3-phosphate (Glyc3P) abolishes the enzyme activity of cardiac aspartate aminotransferase (cAAT). We also examined the protective effects of carnosine against Glyc3P-induced loss of enzyme activity. The present study looked at carnosine's prevention of Glyc3P induced change in protein structure. Purified cAAT (2 mg protein/mL) was incubated with various concentrations of carnosine (1-20 mM) in the presence of Glyc3P (500 microM) for 4 days at 37 degrees C. Following incubation, samples were analyzed by SDS-polyacrylamide gel electrophoresis. Carnosine showed prevention of protein modification at carnosine-to-Glyc3P ratios of 10:1 or greater. There was a progressive loss of the unmodified cAAT protein band as Glyc3P concentration was increased. Additionally, the gel position of the Glyc3P modified cAAT protein varied over time. The apparent molecular weight (MWapp) of the Glyc3P-modified cAAT protein that formed after 1 day at 37 degrees C (500 microM) was greater than its MWapp after 2 days, suggesting that a chemical rearrangement of the initial adduct occurs. These observations support the hypothesis that carnosine is an antiglycation agent and that its mechanism of action involves prevention of protein modification. PMID- 10789501 TI - Characterization of nicotinic acetylcholine receptors from the insects Aphis craccivora, Myzus persicae, and Locusta migratoria by radioligand binding assays: relation to thiamethoxam action. AB - Thiamethoxam, a new neonicotinoid insecticide acting at nicotinic acetylcholine receptors, was characterized in competition binding assays with [3-H] imidacloprid, a specific nicotinic ligand, using membranes from the aphids Aphis craccivora and Myzus persicae, and from the locust Locusta migratoria. In all insects, Scatchard analysis suggested two binding sites for imidacloprid with Kd values in the range of 1 nM and 10 nM, respectively. The Hill values were significantly below 1 (range of 0.63 to 0.85). In contrast to imidacloprid and nicotine, the potency of thiamethoxam to displace [3-H]-imidacloprid varied considerably among these insects. Thiamethoxam was more active than nicotine on Aphis receptors but 100-fold less in Locusta, a nontarget insect. Comparable relations were found to nithiazine. In Myzus, the inhibition curve for thiamethoxam was shallow. This suggested a heterogeneous receptor population displaying a range of binding affinities to thiamethoxam in this aphid. In all three insects, the other neonicotinoid insecticides studied competed with [3-H] imidacloprid in the same order: thiacloprid > imidacloprid > or = acetamiprid > nitenpyram. N-Methylation of imidacloprid strongly reduced the affinity to the imidacloprid site, whereas N-demethylation of thiamethoxam resulted in a comparable increase of affinity. Supplementary assays were performed with (-)-[3 H]-nicotine and [3-H]-alpha-bungarotoxin on locust membranes. Overall, the data suggested that the outstanding insecticidal properties of thiamethoxam may be due to either a different binding site on nicotinic receptors, or receptor isoforms, or specific pharmakokinetic behavior, rather than to exceptional affinity to one of the examined binding sites. PMID- 10789502 TI - Seroepidemiological investigation of feline chlamydiosis in cats and humans in Japan. AB - The prevalence of chlamydia antibodies in Japan was investigated in 215 cat sera, consisting of 88 sera of stray cats and 127 sera of pet cats, and 2,184 human sera, taken from 2,003 general persons and 181 small animal clinic veterinarians, by microimmunofluorescence (MIF) testing with Chlamydia psittaci Fe/Pn1 of feline origin and Prk/6BC of avian origin as antigens. The prevalence rates of anti Fe/Pn1 antibodies were 45.5% in stray cats, 17.3% in pet cats, 1.7% in general persons and 8.8% in small animal clinic veterinarians. The prevalence rates of anti-Prk/6BC antibodies were 51.1% in stray cats, 15.0% in pet cats, 3.1% in general persons and 5.0% in small animal clinic veterinarians. These results suggested that feline chlamydia infection is widely spread in cats especially in stray cats in Japan, and suggested that feline chlamydiosis could be transmitted to people who are in close contact with infected cats. PMID- 10789503 TI - The complete DNA sequence of the O antigen gene region of Plesiomonas shigelloides serotype O17 which is identical to Shigella sonnei form I antigen. AB - We cloned and determined the sequence of a DNA region of approximately 15-kb containing the cluster of genes required for O17 antigen expression in the Escherichia coli K-12 strain from the chromosome of Plesiomonas shigelloides serotype O17:H2 strain. The sequencing analysis revealed that the minimum essential region of the P. shigelloides O17 antigen gene cluster had a size of approximately 11.5-kb and contained 9 contiguous open reading frames (ORFs), which were almost identical to the corresponding ORFs of Shigella sonnei form I antigen gene region, except for IS630 sequence, at the DNA as well as amino acid levels. The putative function of most of the ORFs could be determined on the basis of amino acid sequence similarities and characteristics. In addition, the G+C content of the P. shigelloides O17 antigen genes was lower than that of the chromosomal DNA of P. shigelloides and S. sonnei, suggesting that both P. shigelloides O17 and S. sonnei form I antigen genes had been derived from the same origin with a low G+C content. PMID- 10789504 TI - Phylogenetic analysis of the small hydrophobic (SH) gene of mumps virus in Korea: identification of a new genotype. AB - Viral RNAs extracted from fifteen mumps virus isolated from throat swab, saliva, blood, urine or CSF during mumps epidemics between 1997-1998 in Korea were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and compared by nucleotide sequencing of the small hydrophobic (SH) gene. The deduced amino acid sequences of the SH gene were aligned with the published sequences of mumps virus isolated in different geographic areas. A comparison of the SH gene of mumps viruses in Korea indicated 96.2-100% and 91.2-100% similarity at the nucleotide and amino acid levels, respectively. Phylogenetic analysis, using the neighbor-joining method, showed that Korean mumps virus strains formed a genetically distinct monophyletic group from previously reported genotypes based on the 315-bp length nucleotide and 57 deduced amino acid sequences of the SH gene, and possibly be designated as a new genotype (I). PMID- 10789505 TI - Severity-related molecular differences among nineteen strains of dengue type 2 viruses. AB - Comparative nucleotide sequencing was carried out on dengue type 2 virus (DEN-2) strains isolated from patients in Northeast Thailand during the epidemic season in 1993. The patients exhibited different clinical manifestations ranging from dengue fever (DF) to dengue haemorrhagic fever (DHF)/dengue shock syndrome (DSS). The results classified 19 DEN-2 strains into 3 subtypes according to nonsynonymous amino acid replacements. The strain isolated from a DSS patient eliciting secondary serological response belonged to subtype I, whereas 13 strains isolated from DHF patients with secondary response and 2 strains from DF patients with primary response belonged to subtype II. On the other hand, 3 strains isolated from DF cases evoking either primary or secondary response belonged to subtype III. These results suggest that subtype III virus infection could result in clinically milder manifestation irrespective of the serological response compared with subtype I or II viruses. The RNA secondary structure predicted for the 3' noncoding region showed 4 different structures (A, B, C, and D). The result also indicates that different subtypes of DEN-2 serotypes are circulating in a single epidemic in Thailand. PMID- 10789506 TI - An exfoliative toxin A-converting phage isolated from Staphylococcus aureus strain ZM. AB - Exfoliative toxin A (ETA) causes staphylococcal scalded-skin syndrome in children. The gene for ETA was believed to be coded by the chromosomal DNA. We isolated temperate phages from an ETA-producing strain, ZM, using a restriction minus strain, 1039, as an indicator. One of the prophages, designated phi-ZM-1 mediated lysogenic conversion of ETA. The polymerase chain reaction assay of the eta gene revealed that phage phi-ZM-1 carries the structural gene for ETA. PMID- 10789507 TI - Disintegration of Mg2+ -induced hexagonal assembly of an R-form lipopolysaccharide from Klebsiella pneumoniae by treatment with CaCl2. AB - R-form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN-111 (O3-: K1-), which was precipitated by the addition of 2 volumes of ethanol containing 10 mM MgCl2 for the purification process, ultrastructurally exhibited membrane pieces consisting of an ordered hexagonal lattice structure with a lattice constant of 14 to 15 nm. When the R-form LPS was suspended in 50 mM tris (hydroxymethyl) aminomethane buffer (at pH 8.5) containing 1 mM or higher concentrations of CaCl2 and kept at 4 C for 10 hr, the ordered hexagonal lattice structure of the R-form LPS was disintegrated and changed to an irregular rough, mesh-like structure. By treatment with CaCl2, the content of Mg in the LPS was markedly decreased, and conversely, the content of Ca was increased to a level depending upon the concentration of CaCl2. Results indicate that the addition of CaCl2 to suspensions of the Mg-bound R-form LPS result in a tighter binding of Ca2+ to the R-form LPS and the release of Mg2+ from the R-form LPS, and as a consequence, destroys the Mg2+ -induced ordered hexagonal lattice structure of the R-form LPS. PMID- 10789508 TI - Exacerbation of vascular endothelial injury in the generalized Shwartzman reaction by the administration of anti-E-selectin antibody. AB - Previously, we reported that the consecutive administration of lipopolysaccharide (LPS) into LPS-sensitized mice for the generalized Shwartzman reaction (GSR) induced systemic injury of vascular endothelial cells. The aim of this study was to investigate the participation of vascular adhesion molecules in the vascular endothelial injury of GSR. The administration of anti-E-selectin antibody in GSR induced mice resulted in massive apoptosis of vascular endothelial cells and congestion in blood vessels. Further, marked hemorrhage was found in the pulmonary alveoli of those mice. GSR, especially lung injury, was definitely exacerbated by the administration of anti-E-selectin antibody. On the other hand, the administration of anti-VCAM-1 antibody did not induce such injury of vascular endothelial cells. The possible role of E-selectin in the exacerbation of vascular endothelial injury in GSR is discussed. PMID- 10789509 TI - Isolation of Orientia tsutsugamushi from Leptotrombidium fuji and its characterization. AB - In our attempts to isolate Orientia tsutsugamushi from trombiculid mites, a strain was successfully isolated from Leptotrombidium fuji collected in Aichi Prefecture, Japan. This is the first case of isolation of O. tsutsugamushi from L. fuji. A phylogenetic analysis based on the base-sequence homology of the 56 kDa type-specific antigen-gene indicated that the strain is a new type which is not closely related to any strains analyzed previously. Three strains isolated from Leptotrombidium pallidum harvested at the same area were identified as being closely related to the JP-2 type (subtype-2 of Karp type in Japan) by phylogenetic analysis. PMID- 10789510 TI - Inhibition of HIV-1 infection by an intramolecular antisense peptide to T20 in gp160. AB - Antisense amino acids are amino acids which can be translated from the corresponding anti-codons of a sense amino acid. Antisense peptides encoded by the noncoding DNA strand have a tendency to interact with each other. We have demonstrated that antisense peptide sequences are present intramolecularly, and these may contribute to the folding and maintenance of the tertiary structure of a protein. T20 is a synthetic peptide with an amino acid sequence in the gp41 of HIV-1 and has been demonstrated to be a potent inhibitor of HIV-1 infection. We searched for intramolecular peptide sequences which are antisense to portions of T20. A synthetic peptide (TA-1L) consisting of amino acids 84 to 97 of gp160, which contains an antisense peptide sequence (TA-1) to T20, was shown to inhibit HIV-1(IIIB) infection of MT-4 cells. Interaction of these antisense peptides could be involved in sustaining HIV-1 infectivity. The TA-1L site, which exists in the C1 domain of gp160, is highly homologous among strains of HIV-1, especially at TA-1 and in the amino acids flanking the C terminus. Although the TA-1 sites of 18 out of 30 HIV-1 strains were antisense to the T20 region, those of the remaining 12 strains, including HIV-1(MN), were not. However, TA-1L inhibited infection by HIV-1(MN), which has no antisense peptide in T20 corresponding to TA-1, although the inhibitory effect was weaker. TA-1L may thus also interfere with the gp160 interaction with CD4, which has an antisense sequence to TA-1. PMID- 10789511 TI - Veterinary public health: past, present, and future. PMID- 10789512 TI - Bone disorders in the dog: a review of modern genetic strategies to find the underlying causes. AB - In man, the genetic defects of more than 600 inherited diseases, of which at least 150 skeletal diseases, have been identified as is the chromosomal location for approximately 7000 genes. This rapid progress has been made possible by the generation of a genetical and physical map of the human genome. There is no reason to believe that for the dog not a similar development may occur. This review is therefore focussed on the use of novel tools now available for comparative molecular genetic studies of skeletal dysplasias in the dog. Because the genomes of mammals at the subchromosomal level are very well conserved, likely candidate disease genes known from other species might be considered. In this review, formation of the bones and the most important canine disorders of the skeleton influencing locomotion will be discussed first. The canine disorders discussed are canine hip dysplasia, the three different forms of elbow dysplasia (fragmented coronoid process, ununited anconeal process, osteochondrosis dissecans and incongruency) and dwarfism. Where possible a link is made with similar diseases in man or mouse. Then, the molecular biological tools available to analyse the genetic defect will be reviewed and some examples discussed. PMID- 10789514 TI - Comparison of two different meat inspection techniques. AB - Field studies of meat inspection procedures in three different slaughter houses were performed in Germany, in 1996. In total 22,634 fattening pigs from 63 farms were inspected twice, using a visual and the traditional (mandatory inspection system) procedure. Statistical analysis was performed using the rate of ND+ (Non Detected Positives) for both visual and traditional inspection. Neither the visual nor the traditional methods detected all lesions which were supposed to be in the lot of animals inspected. Some of the results could be explained by the nature and obviousness of the respective lesion. Conclusions regarding the future performance of inspection should not be drawn before a formal risk analysis procedure for the particular lesion has been done. PMID- 10789513 TI - Diagnosis and epidemiology of bovine neosporosis: a review. AB - Neospora caninum is a protozoan parasite that was first described in dogs in 1988. Subsequently the parasite has been associated with abortion in cattle. Evidence is accumulating that neosporosis is a major cause of reproductive failure in cattle all over the world. Recent knowledge of neosporosis is reviewed with emphasis on the diagnosis and epidemiology of the disease in cattle. In addition, attention is paid to strategies for disease control and prevention. PMID- 10789515 TI - An equine herpesvirus 1 (EHV1) abortion storm at a riding school. AB - An outbreak of EHV1 abortions occurred at a riding school in The Netherlands in 1991. Seven of twelve pregnant mares aborted, and another foal died at 8 days of age. Six abortions occurred within 12 days in March after an initial abortion on 8 February. Four mares delivered live foals. Virological examination of four aborted foals revealed an EHV1 infection. Serological results for paired sera from 17 horses suggested, that the initial abortion on 8 February was the index case, and probably caused the other six abortions. The index case could well have been caused by reactivation of latent virus induced by transport stress. The laboratory results are discussed in the light of the present knowledge of the pathogenesis and epidemiology of EHV1 abortion. PMID- 10789516 TI - Differentiation and genomic and antigenic variation among fetal, respiratory, and neurological isolates from EHV1 and EHV4 infections in The Netherlands. AB - Ten monoclonal antibodies (MAbs) were produced against equine herpes virus type 1 (EHV1). Two appeared type-specific, while the other eight were directed against epitopes common to both EHV1 and EHV4. Two MAbs directed against the glycoprotein gp2 recognized linear epitopes, as demonstrated by Western blotting. With pools of type-specific MAbs, 282 field isolates were typed in an immunoperoxidase monolayer assay (IPMA). From a total of 254 fetal or neonatal isolates, 244 (96%) were typed as EHV1, whereas 14 out of 15 (93%) respiratory tract isolates were typed as EHV4. Surprisingly, 3 out of 13 isolates (23%) originating from horses with neurological disease were typed as EHV4. No antigenic differences were found among 75 randomly selected EHV1 field isolates, using the panel of ten MAbs and six additional MAbs, directed against gp2, gB, or gC. Typing by restriction endonuclease analysis with BamHI corresponded completely with that of MAb analysis. There was a remarkable degree of uniformity in BamHI restriction patterns, with 90% of the investigated EHV1 isolates belonging to the 1P electropherotype. Among 30 randomly selected EHV1 isolates we could not identify the EHV1.1B electropherotype, which has been the predominant electropherotype in Kentucky since 1982. Mobility differences were seen in fragments originating from the repeat regions. These differences were not caused by heterologous cell passage, since all viruses were passaged in equine cell systems. PMID- 10789517 TI - Gauged attenuation of congenital portosystemic shunts: results in 160 dogs and 15 cats. AB - Portosystemic shunts were ligated over a gauged stainless steel rod in 160 dogs and 15 cats, using a midline celiotomy. The diameter of the rod varied with the size of the shunt and the diameter of the portal vein cranial to the shunt. Shunts were narrowed to the smallest diameter that did not cause signs of portal hypertension such as cyanosis of the stomach, pancreas, and small intestine. A slight discoloration was accepted only if the heart rate, end-expiratory CO2%, or arterial blood pressure (if available) did not deviate more than 15% from the values that were recorded at the beginning of the surgical procedure. The perioperative mortality (0-30 days) was 29%. The most common cause of death was euthanasia because of hypoplasia of the portal vein cranial to the shunt. Animals with intrahepatic shunts had a significantly lower probability of survival than animals with extrahepatic portocaval or portoazygos shunts. In dogs, large breed and a high body weight were also significant risk factors for non-survival. Age had a significant effect on risk of non-survival, with an increased risk for older dogs, irrespective of the breed of the dog (large breed vs. small breed). The probability of survival without recurrence of hepatoencephalopathy (HE) after 1 and 4 years was 61.3% and 55.7%, respectively. The only variable that was significantly associated with non-recurrence of HE was the breed of the dog, there being a lower probability for large breeds. Among the animals that survived surgery for more than 30 days, there was a significant higher probability of recurrence of HE in cats than in dogs. PMID- 10789518 TI - The serological BHV1 status of dams determines the precolostral status of their calves. AB - Precolostral calves and their dams were serologically investigated for the presence of antibodies against Bovine Herpesvirus 1 in diagnostic tests with a very high sensitivity and specificity. Although the syndesmo-chorial type of placenta of ruminants does not transfer gamma globulins, a large number of calves had antibodies, in most cases in a very low concentration. Significant correlations were found between the serological status of the dam, the status of the calf, and the titre of antibodies. Oral intake of maternal blood by the calf at birth or transmission or leakage of maternal antibodies during pregnancy might be possible causes of precolostrally positive calves. From the results it is concluded that to reduce the risk of obtaining BHV1-positive calves, BHV1 negative dams should be selected for breeding purposes. PMID- 10789519 TI - Aujeszky's disease virus eradication campaign successfully heading for last stage in The Netherlands. AB - The Aujeszky's disease virus (ADV) eradication campaign in The Netherlands is based on compulsory vaccination and certification of ADV-free herds and consists of three stages: reduction of transmission of ADV, tracing and elimination of remaining sources of ADV, and, in the last stage prohibition of vaccination. This paper summarizes the progress made in the first two stages of the eradication. Between 1994-1999, ADV seroprevalence decreased sharply in all four regions in The Netherlands, both in the sow and the finishing pig population. In the sow population, the mean ADV prevalence in the northern, eastern and western regions decreased to less than 0.5% in the first quarter of 1998, and to approximately 1% in the southern region. For the finishing pig population, the mean ADV prevalence decreased to less than 0.6% in all four regions in the first quarter of 1998. The proportion of sow herds that were certified ADV-free increased steadily from approximately 40% in the last quarter of 1996 to 96% in the first quarter of 1999. The proportion of finishing pig herds that were certified ADV-free showed the same spectacular increase as in the sow population: from approximately 14% in the last quarter of 1996 to 90% in the first quarter of 1999. It is anticipated that the last stage of the ADV eradication campaign, stopping vaccination, will be achieved very soon. Monitoring wild boars for ADV between 1994 and 1999 indicated that ADV is uncommon within the wild boar population. Therefore, it seems that ADV infection in the wild boar population is not an important reservoir in The Netherlands. PMID- 10789520 TI - Pyothorax in nine dogs. AB - The results of treatment of pyothorax using systemic antibiotics, drainage, and lavage of the pleural space, are reported for 9 dogs. All 9 dogs recovered completely. In 8 of the 9 dogs the follow-up period was at least 6 months and in none was there a relapse. The results obtained with this treatment are excellent in comparison with the results that have been reported for treatment with systemic antibiotics and drainage of the pleural space but without lavage. Apart from the addition of pleural lavage to the treatment protocol, the better result might be because migrating plant related foreign bodies did not seem to play an important role in the pathogenesis of pyothorax in this group of dogs. PMID- 10789521 TI - Assessment of health risks of large semi-wild herbivores in urbanized areas. AB - The health risks for both domestic animals and humans caused by large herbivores in self-sustaining ecosystems are largely unknown. The aim of this article is to make an inventory of these risks, to explore ways to manage them in practice, and to make recommendations for the quantification of risks. Potential hazards from herbivores in and around Europe are listed using the data of the OIE (Office International des Epizooties). The desired health status and the implementation of control or surveillance measures are important factors when assessing the risks. Results indicate that a regular yearly system of health monitoring of herbivores is necessary. To get more insight into the importance of certain risks (Infectious Bovine Rhinotracheitis, biodegradation of carrion in the field) epidemiological investigations have to be carried out to assess the risk of transmission in different situations (with or without intervention). Analysing and managing risks enable decision-makers to formulate the conditions for the development of nature reserves. In Europe more has to be done to increase the quality of nature in terms of de-fragmentation and de-isolation, but regulations concerning the health of large herbivores also have to be improved. PMID- 10789522 TI - Flow cytometric analysis of the DNA content of bovine and human bone marrow cells. AB - The defence against infection in high-yielding dairy cows is correlated with the number and function of circulating neutrophils and depends on their production in bone marrow. Therefore, the DNA content of isolated bone marrow cell suspensions from 7 calves, 7 cows and 14 humans was assayed by flow cytometry. Bovine sternal bone marrow samples were collected within 30 min of death, and human marrow samples were collected by sternal puncture and aspiration. Mononucleated cells were isolated by gradient centrifugation. In the bone marrow samples from calves and cows, 35 +/- 2.6% and 31.8 +/- 1.5% of the isolated bone marrow cells respectively were in the S/G2/M-phase. The difference between calves and cows was not significant. In the human samples, only 12 +/- 0.8% of the cells were in the S/G2/M-phase. A significant (P < 0.001) difference was observed between the two species. These results indicated that the proliferative, in activity of haematopoietic cells is significantly higher in cattle than in humans. PMID- 10789523 TI - Introduction: the Australian mental health survey. PMID- 10789524 TI - Australia's mental health: an overview of the general population survey. AB - OBJECTIVES: The objectives of this study were to estimate the 1-month and 1-year prevalence of mental disorders in the Australian adult population; to determine the amount of disablement associated with this; and to determine the use of health and other services by persons with common mental disorders. METHOD: For the Adult Survey, a household sample of 10600 persons aged 18 years and over were interviewed across Australia by experienced field staff of the Australian Bureau of Statistics. This was 78% of the target sample. The interview consisted of the composite international diagnostic interview in its automated presentation (CIDI A) and other components to determine disablement, use of services and satisfaction with services received. The diagnostic classifications used in the analyses were both ICD-10 and DSM-IV. Only the results from ICD-10 are reported here. RESULTS: A total of 17.7% of the sample had one or more common mental disorders, anxiety, depression, alcohol or substance abuse and neurasthenia. This morbidity was associated with considerable disablement in daily life: 3 days of impaired social role performance in the previous 4 weeks, compared with 1 day for the general population. Of all cases, 64.6% had had no contact with health services in the previous year; 29.4% had seen GPs and 7.5% had seen psychiatrists. CONCLUSION: Australia now has its own national estimates of psychiatric morbidity. The morbidity is associated with considerable disablement, but most of it is untreated. General practitioners encounter by far the largest proportion of those reaching services. PMID- 10789525 TI - Alcohol- and drug-use disorders in Australia: implications of the National Survey of Mental Health and Wellbeing. AB - OBJECTIVE: This study reports the prevalence and correlates of ICD-10 alcohol- and drug-use disorders in the National Survey of Mental Health and Wellbeing (NSMHWB) and discusses their implications for treatment. METHOD: The NSMHWB was a nationally representative household survey of 10641 Australian adults that assessed participants for symptoms of the most prevalent ICD-10 and DSM-IV mental disorders, including alcohol- and drug-use disorders. RESULTS: In the past 12 months 6.5% of Australian adults met criteria for an ICD-10 alcohol-use disorder and 2.2% had another ICD-10 drug-use disorder. Men were at higher risk than women of developing alcohol- and drug-use disorders and the prevalence of both disorders decreased with increasing age. There were high rates of comorbidity between alcohol- and other drug-use disorders and mental disorders and low rates of treatment seeking. CONCLUSIONS: Alcohol-use disorders are a major mental health and public health issue in Australia. Drug-use disorders are less common than alcohol-use disorders, but still affect a substantial minority of Australian adults. Treatment seeking among persons with alcohol- and other drug-use disorders is low. A range of public health strategies (including improved specialist treatment services) are needed to reduce the prevalence of these disorders. PMID- 10789526 TI - The National Survey of Mental Health and Wellbeing: the child and adolescent component. AB - OBJECTIVE: This paper describes the Child and Adolescent Component of the National Survey of Mental Health and Wellbeing. METHOD: The aims of the study, critical decisions in planning for the study, progress to date and key issues which influenced the course of the study are described. RESULTS: The Child and Adolescent Component of the National Survey of Mental Health and Wellbeing is the largest study of child and adolescent mental health conducted in Australia and one of the few national studies to be conducted in the world. Results from the study will provide the first national picture of child and adolescent mental health in Australia. CONCLUSIONS: Large-scale epidemiological studies have the potential to provide considerable information about the mental health of children and adolescents. However, having a clear set of aims, ensuring that the scope of the study remains within manageable proportions and paying careful attention to the details of fieldwork are essential to ensure that high-quality data is obtained in such studies. PMID- 10789528 TI - Way out of tune: lessons from Shine and its expose. AB - OBJECTIVE: The depiction of David Helfgott's life presented in the movie Shine is at odds with other public accounts, notably one by his sister, Margaret. These significant discrepancies have sparked a prolonged media debate and provide the opportunity to examine cinema's apparent ground rules governing depictions of psychiatry in film, the media values and pressures which are claimed to limit the scope of these portrayals, and the implications for psychiatry. METHOD: Information was obtained from a number of sources, including Shine, books about the movie and Shine film paraphernalia, other films about mental illness, the psychiatric papers on cinema, media images of mental illness and media values, and through discussions with fellow mental health professionals, consumers, carers and media specialists. RESULTS: David Helfgott emerges as an undoubtedly remarkable and resilient individual, who, together with his family, was vulnerable to, and may have experienced, exploitation and violation through the cinema. CONCLUSIONS: Filmmakers should reconcile media values and constraints with considerations of ethics and public accountability. Marrying these considerations is both possible and compatible with good filmmaking and audience appeal. There is the potential for a story about those who have mental illness to be told from multiple points of view without compromising dramatic power. PMID- 10789527 TI - Psychotic disorders in urban areas: an overview of the Study on Low Prevalence Disorders. AB - OBJECTIVE: This paper reports on a study designed within the framework of the National Survey of Mental Health and Wellbeing to: estimate the prevalence of psychoses in urban areas of Australia; identify profiles of symptomatology, impairments and disabilities; collect information on services received and needed; and explore quality of life issues in a broadly representative sample of people with psychotic illnesses. METHOD: The study was conducted over four areas in the Australian Capital Territory, Queensland, Victoria and Western Australia, as a two-phase survey: (i) a census and screening for psychosis of all individuals who made contacts with mental health services during a period of 1 month in 1997; and (ii) interviews with a stratified random sample (n=980) of the screen-positive individuals (n=3800) using a standardised instrument. RESULTS: The point prevalence (1 month) of psychotic disorders in the urban population aged 18-64 is in the range of 4-7 per 1000 with a weighted mean of 4.7 per 1000. People with psychotic disorders experience high rates of functional impairments and disability, decreased quality of life, persistent symptoms, substance-use comorbidity and frequent side effects of medication. Although the utilisation of hospital-based and community mental health services, as well as of public and non governmental helping agencies, is high, the majority live in extreme social isolation and adverse socioeconomic circumstances. Among the many unmet needs, the limited availability of community-based rehabilitation, supported accommodation and employment opportunities is particularly prominent. CONCLUSIONS: The so-called 'low-prevalence' psychotic disorders represent a major and complex public health problem, associated with heavy personal and social costs. There is a need for a broad programmatic approach, involving various sectors of the community, to tackle the multiple dimensions of clinical disorder, personal functioning and socioeconomic environment that influence the course and outcome of psychosis and ultimately determine the effectiveness of service-based intervention. PMID- 10789529 TI - The internal and external boundaries of obsessive-compulsive disorder. AB - OBJECTIVE: This study aims to explore the internal and external boundaries of obsessive-compulsive disorder (OCD). METHOD: Selected literature is reviewed. RESULTS: Prevalence and incidence estimates for OCD vary markedly according to diagnostic criteria applied. External boundaries of the disorder are permeable, with a large degree of overlap with other psychiatric disorders, and an association with certain neurological disorders. Some cases of OCD appear to have a neurodevelopmental origin. CONCLUSIONS: Further delineation of disorders characterised by obsessional and compulsive symptoms can inform models of aetiology, pathophysiology and treatment. PMID- 10789531 TI - Therapeutic day programs in the treatment of adolescents with mental illness. AB - OBJECTIVES: This study aims to assess the effectiveness of day programs against other treatment modalities in adolescents with mental illnesses (including substance abuse). It also aims to identify patient and program factors associated with favourable outcomes for adolescents with mental illness treated in therapeutic day programs. METHOD: An extensive database search of published literature was conducted, supplemented by hand-searching of references of retrieved articles, and a search by author. There was no restriction by study design. Relevant studies were identified and appraised using standardised criteria. RESULTS: Of the 231 studies identified in the search, 26 were relevant to this review. The majority of studies identified day programs as effective. There was a lack of adequate comparisons between day programs and other organisational arrangements. CONCLUSIONS: The quality of the studies identified was low. Twenty-two of the 26 studies did not have an adequate control group. Higher-quality research is required in the evaluation of the effectiveness of day programs, and the identification of other factors promoting successful outcome in the treatment of adolescent mental illness. PMID- 10789530 TI - Lesbians in New Zealand: their mental health and satisfaction with mental health services. AB - OBJECTIVES: To describe the mental health of lesbians in New Zealand, and to document their accounts of their experience of mental health services. METHOD: This is a descriptive cross-sectional study. A postal questionnaire, the Lesbian Mental Health Survey, was distributed via lesbian newsletters to 1222 women throughout New Zealand. Mental health measures included the General Health Questionnaire (GHQ-28), Interview Schedule for Social Interaction (ISSI), and respondents' histories of sexual abuse and psychiatric histories. Experiences of mental health services were sought. RESULTS: The estimated response rate was 50.8%. The respondent group were predominantly New Zealand European, highly educated, urban women between 25 and 50 years of age. Three-quarters had identified as lesbian for more than 5 years. Recent self-identification as lesbian was associated with higher GHQ score, as was being younger than 35, having a history of sexual abuse, and not living with a partner. Eighty percent of respondents had used mental health services sometime in their lives and nearly 30 percent of users had received 'lesbian-unfriendly' treatment at some point. One-sixth of respondents had experienced discrimination from service providers in the previous 5 years. CONCLUSION: While the mental health of lesbians is influenced by factors similar to those influencing women's mental health in general, because of social factors, such as stigma and isolation, lesbians may be more vulnerable to common mental illnesses. Health professionals, mental health professionals in particular, need to raise their awareness of the issues lesbians face in dealing with their sexuality, therapeutic relationships and mental health services. Increased training about sexuality for health professionals, as well as further research into areas such as stress and stigma, sexual abuse and attempted suicide among lesbian women, is recommended. PMID- 10789532 TI - Obstacles to a randomised controlled trial of intensive dynamic psychotherapy: an account of the New South Wales Section of Psychotherapy outcomes project. AB - OBJECTIVE: This paper examines the obstacles to a randomised controlled trial (RCT) of intensive dynamic psychotherapy (IDP) by reference to the fate of the New South Wales Section of Psychotherapy outcomes project. METHOD: Planning was complete and the final research protocol was about to be implemented when funding difficulties led to suspension of the project. The opinions of the research subcommittee regarding the main obstacles to the ultimate success of the project are now analysed in the expectation that better research strategies will follow. RESULTS: With hindsight, six of the eight members of the research subcommittee reported that the project was not feasible. By choice of questionnaire items they identified the greatest threats to a successful trial as: standardisation of the procedures, termination at 24 months, the availability of funding and the choice of treatment procedures. The most frequently volunteered concerns related to the enlistment and cooperation of the trial therapists (5), standardisation of the experimental therapy (3), probable shortfall in trial subjects (3) and the availability of funding (2). CONCLUSIONS: The most powerful general obstacles to success of the project related to the standardisation of procedures and the failure to maintain sufficient cooperation of trial therapists. The protocol required IDP therapists to terminate procedures at 24 months, which contradicted their usual practices and led to some alienation from the project. Amendments to the protocol might improve the possibility of a successful trial. However, one might also conclude that it is premature to attempt a naturalistic RCT of IDP. PMID- 10789533 TI - Contrasting views and experiences of health professionals on the management of comorbid substance misuse and mental disorders. AB - OBJECTIVES: To determine opinions and experiences of health professionals concerning the management of people with comorbid substance misuse and mental health disorders. METHOD: We conducted a survey of staff from mental health services and alcohol and drug services across Queensland. Survey items on problems and potential solutions had been generated by focus groups. RESULTS: We analysed responses from 112 staff of alcohol and drug services and 380 mental health staff, representing a return of 79% and 42% respectively of the distributed surveys. One or more issues presented a substantial clinical management problem for 98% of respondents. Needs for increased facilities or services for dual disorder clients figured prominently. These included accommodation or respite care, work and rehabilitation programs, and support groups and resource materials for families. Needs for adolescent dual diagnosis services and after-hours alcohol and drug consultations were also reported. Each of these issues raised substantial problems for over 70% of staff. Another set of problems involved coordination of client care across mental health and alcohol and drug services, including disputes over duty of care. Difficulties with intersectoral liaison were more pronounced for alcohol and drug staff than for mental health. A majority of survey respondents identified 13 solutions as practical. These included routine screening for dual diagnosis at intake, and a range of proposals for closer intersectoral communication such as exchanging client information, developing shared treatment plans, conducting joint case conferences and offering consultation facilities. CONCLUSIONS: A wide range of problems for the management of comorbid disorders were identified. While solution of some problems will require resource allocation, many may be addressed by closer liaison between existing services. PMID- 10789534 TI - Costing depression and its management: an Australian study. AB - OBJECTIVE: To examine the cost impact of referral to a Mood Disorders Unit (MDU), by comparing pre-service and post-service costs, and MDU and control samples. METHOD: We studied tertiary referral MDU patients and a control group of consultants' depressed patients, with the principal comparison intervals being: (i) 12 months prior to and (ii) 6 months following baseline assessment, with costs annualised to allow the impact of assessment and treatment recommendation to be determined. In addition, we assessed any 'personal cost' of depression. RESULTS: Following baseline assessment, MDU referrals showed a reduction in costs, while controls' costs increased, largely driven by contrasting directions in hospitalisation and social welfare costs. We identify variables associated with high and increased costs, including features of the earlier stages of the disorder, whether social welfare was received, diagnostic subtype and personality dysfunction, with multivariate analyses refining the variable sets. Self-report data indicated that patients judged the 'personal cost' of depression to exceed more formal cost parameters, so that to experience depression is itself depressogenic. CONCLUSIONS: This first Australian attempt to cost depression and its management in the clinical setting more provides a methodology for wider application in service evaluation studies rather than delivers an unequivocal answer to whether a specialist Mood Disorders Unit is cost efficient or not. PMID- 10789535 TI - Cognitive-behaviour therapy as a treatment for irritable bowel syndrome: a pilot study. AB - OBJECTIVE: The irritable bowel syndrome (IBS) is a chronic and often disabling functional bowel disorder. Psychological treatments, in particular cognitive and behavioural interventions, have been shown to be effective for this disorder. The aim of this study was to test the efficacy of a cognitive-behaviour program. METHOD: Eight participants (seven female, one male) aged between 24 and 71 years, with a diagnosis of IBS according to the Rome criteria, were recruited from among the gastroenterology outpatients at Nepean Hospital, in Sydney, Australia. Participants were administered pretreatment on measures of psychological function and bowel symptom severity. Following a 2-week baseline period, participants began a structured psychological treatment comprising eight sessions of cognitive behaviour therapy. Throughout treatment, participants maintained daily records of symptom severity and completed homework assignments to ensure treatment compliance. The pretreatment assessment measures were repeated 1 week post treatment. RESULTS: After treatment, five of the eight patients no longer met the Rome diagnostic criteria for IBS. There was no significant reduction in bowel symptom frequency. There were, however, significant improvements in the distress and disability associated with bowel symptoms. Anxiety and depression were also significantly reduced. CONCLUSIONS: Cognitive-behaviour therapy reduced the distress and disability associated with IBS, but not the frequency of bowel symptoms. This supports the proposed cognitive model for IBS, and cognitive behaviour therapy appears to have its effect by altering the cognitive response to visceral hypersensitivity. PMID- 10789536 TI - Antidepressants versus placebo for the treatment of bulimia nervosa: a systematic review. AB - OBJECTIVE: The objective of this study was to valuate the effectiveness, tolerability and acceptability of various classes of antidepressants compared with placebo in the treatment of bulimia nervosa. METHOD: A meta-analysis including 16 randomised controlled trials and 1300 bulimic patients was performed. Dichotomous outcomes were analysed by calculating relative risks, and continuous outcomes by calculating effect sizes. Methodological quality, heterogeneity in the results and selective publication were assessed. RESULTS: Short-term remission in bulimic symptoms was statistically more likely on antidepressants than placebo (Relative Risk=0.88, 95% CI=0.83-0.94, p<0.0001). Drop-out rates were high but no statistical difference was found between treatment groups (34.6% and 31.4% for drug and placebo; RR=1.03, 95% CI=0.80 1.32, p=0.8). No difference in efficacy could be demonstrated among different classes of antidepressants. CONCLUSIONS: The use of a single antidepressant agent was clinically effective for the treatment of bulimia nervosa when compared with placebo, with an overall greater remission rate and a higher rate of drop-outs. No differential effect regarding efficacy and tolerability among the various classes of antidepressants could be demonstrated. PMID- 10789537 TI - Medicine, psychiatry and euthanasia: an argument against mandatory psychiatric review. AB - OBJECTIVE: The paper critically appraises the argument that requests for active assistance to die should be subject to mandatory psychiatric assessment. METHOD: The argument for mandatory psychiatric assessment is usually supported by an appeal to the need for safeguards against errors and omissions in both the diagnosis of psychiatric conditions affecting the terminally ill and the exploration of the meanings of their requests. This intuitively appealing view is challenged through a broader analysis which examines connections between medicine's traditional adherence to the moral distinction between acts and omissions and the following issues: the historical relationship between medical practice and dying, the recent development of research into treatment-withdrawal decisions, the scientific status of psychiatry, the logic of rationality and decision-making competence. RESULTS: The analysis reveals a number of hitherto unexamined and unacknowledged influences which would make psychiatric review of requests for assisted death a much less objective and impartial process than is assumed. CONCLUSION: Mandatory psychiatric review is an instance of the medicalisation of death and dying which could abridge the freedom of certain individuals to make decisions about their deaths. PMID- 10789538 TI - Demoralisation, depression and desire for death: problems with the Dutch guidelines for euthanasia of the mentally ill. AB - OBJECTIVE: Given the recent publication of Dutch guidelines for euthanasia of the mentally ill, our aim in this paper is to review the clinical role of the psychiatrist in assessing patients who seek euthanasia or physician-assisted suicide. METHOD: Three areas are examined that are informative of the role of the psychiatrist in assessing patients who desire death: recent surveys of psychiatrists' attitudes, empirical and clinical studies, and treatment issues. RESULTS: Demoralisation and depression emerge as pertinent clinical issues worthy of psychiatric intervention. The role of the psychiatrist is complex and includes issues of assessment, systems, countertransference and treatment. CONCLUSIONS: Dutch guidelines for physician-assisted suicide in the mentally ill generate serious concern given the uncertainty of prognosis, potential range and variability of outcome of treatments of suicidality and the boundary violations that are involved for the psychiatrist. The guidelines have the potential to dangerously alter the practice of psychiatry and should be condemned. PMID- 10789539 TI - Nonconvulsive generalised status epilepticus following electroconvulsive therapy. AB - OBJECTIVE: To describe a case of nonconvulsive generalised status epilepticus (NGS) following electroconvulsive therapy (ECT). CLINICAL PICTURE: A 40-year-old woman suffering from a major depressive episode was treated with ECT following treatment with clonazepam, haloperidol and paroxetine. After her fifth treatment she became acutely confused. An electroencephalogram (EEG) at the time was consistent with NGS. TREATMENT AND OUTCOME: Initially intravenous diazepam and phenytoin were administered with an improvement in both her mental state and EEG. An oral anticonvulsant was continued. CONCLUSIONS: NGS is a rare though treatable cause of confusional states following ECT and should be considered in the differential diagnosis. PMID- 10789540 TI - Criteria for involuntary hospitalisation. PMID- 10789541 TI - Fluoxetine for preoccupation with delusion. PMID- 10789542 TI - Management of acute psychosis. PMID- 10789543 TI - Criteria for neurasthenia. PMID- 10789544 TI - Perhaps not so rare? PMID- 10789545 TI - A rhythmic tic in Tourette's syndrome. PMID- 10789546 TI - The immune system and infection may be involved in the effects of psychological factors on the cardiovascular system. PMID- 10789547 TI - Reply to Boyce and Judd. The place for the tricyclic antidepressants in the treatment of depression. PMID- 10789548 TI - Novel antipsychotic drugs and INH-related psychosis. PMID- 10789549 TI - Antibiotic prophylaxis in children with relapsing urinary tract infections: review. AB - Recurrent urinary tract infections (UTIs) are observed in 30-50% of children after the first UTI. Of these, approximately 90% occur within 3 months of the initial episode. The basic aim of antibiotic prophylaxis in children with malformative uropathy and/or recurrent UTIs, is to reduce the frequency of UTIs. The bacteria most frequently responsible for UTI are gram-negative organisms, with Escherichia coli accounting for 80% of urinary tract pathogens. In children with recurrent UTIs and in those treated with antibiotic prophylaxis there is a greater incidence of UTI due to Proteus spp., Klebsiella spp. and Enterobacter spp., whereas Pseudomonas spp., Serratia spp. and Candida spp. are more frequent in children with urogenital abnormalities and/or undergoing invasive instrumental investigations. Several factors are involved in the pathogenesis of UTI, the main ones being circumcision, periurethral flora, micturition disorders, bowel disorders, local factors and hygienic measures. Several factors facilitate UTI relapse: malformative uropathies, particularly of the obstructive type; vesico ureteric reflux (VUR); previous repeated episodes of cystitis and/or pyelonephritis (3 or more episodes a year), even in the absence of urinary tract abnormalities; a frequently catheterized neurogenic bladder; kidney transplant. The precise mechanism of action of low-dose antibiotics is not yet fully known. The characteristics of the ideal prophylactic agent are presented in this review, as well as indications, dosages, side effects, clinical data of all molecules. While inappropriate use of antibiotic prophylaxis encourages the emergence of microbial resistance, its proper use may be of great value in clinical practice, by reducing the frequency and clinical expression of UTIs and, in some cases such as VUR, significantly helping to resolve the underlying pathology. PMID- 10789550 TI - Short-term bactericidal activity of amoxicillin and cefotaxime against penicillin susceptible and -resistant pneumococcal strains: an in vitro pharmacodynamic simulation. AB - The 8-hour in vitro activity of serum-simulated concentrations of amoxicillin (obtained after 875 mg oral dose) and cefotaxime (obtained after a 1 g i.v. dose), against 20 strains of the 5 Streptococcus pneumoniae serotypes most prevalent in Spain, was explored. Despite a greater initial inocula decrease observed with cefotaxime against the resistant strains at the first sampling time, a decrease > or =99.9% was obtained with both beta-lactams from 6h onwards against the penicillin-susceptible strains; the same was observed for the penicillin-resistant strains with amoxicillin but not with cefotaxime. PMID- 10789551 TI - Effects of subinhibitory concentrations of moxifloxacin in an in-vitro dynamic model. AB - The postantibiotic effect (PAE), sub-MIC effect (SME) and postantibiotic sub-MIC effect (PASME) of moxifloxacin were investigated in an in-vitro dynamic model reproducing in-vivo elimination kinetics of the antibiotic. The PAE was induced by exposing strains of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli and Klebsiella pneumoniae to 5 x MIC of the antibiotic for 1.5 hours. After induction, cultures were washed to eliminate the antibiotic and resuspended into the dynamic model either in the presence or absence of a subinhibitory concentration of the antibacterial agent of 0.5 x MIC. Unexposed controls were treated similarly. PASMEs were constantly longer than corresponding SMEs, but differences between them were not statistically significant. Both PASMEs (mean 11:17 hours, range from 8:17 to 14:57) and SMEs (mean 9:23 hours, range from 6:03 to 12:34) had an initial bactericidal effect and were significantly longer than PAEs (mean 1:31 hours, range from 0:21 to 2:14). The primary effect of moxifloxacin sub-MICs appears to be prevalent in PAE. The possibility of once-daily dosing of the drug is strengthened. PMID- 10789552 TI - Penicillin-resistant and -intermediate Streptococcus pneumoniae in Saudi Arabia. AB - The antibiotic susceptibility was analyzed of approximately 400 consecutive isolates of S. pneumoniae isolated from different regions of Saudi Arabia. Most of these isolates were from respiratory (sputum, otitis, 53.8%), blood/CSF (26.3%) and ophthalmic (20%) specimens. Overall 6.2% of the isolates were penicillin-resistant (MICs > or =2 microg/ml) and 51.2% were -intermediate (MICs 0.1-1 microg/ml). The resistance rates to cefuroxime, clarithromycin and ceftriaxone were 14.9%, 14.8% and 4.5% respectively. Only 3.5% of S. pneumoniae showed resistance to amoxycillin/clavulanic acid. The MICs of all tested antibiotics increased as did the penicillin MICs. Penicillin resistance was significantly associated with resistance to cefuroxime (p<0.001) but not with the others. These data indicate the presence of penicillin and multiple-resistant pneumococci in Saudi Arabia and that these strains can spread among individuals. A greater awareness with extended indications for microbiological diagnosis, antimicrobial susceptibility testing and restrictive prescription of antibiotics are needed. PMID- 10789553 TI - Analysis of a mini-outbreak of methicillin-resistant Staphylococcus aureus in a surgical ward by using arbitrarily primed-polymerase chain reaction. AB - In November 1995, an increase was noticed in the number of methicillin-resistant Staphylococcus aureus (MRSA) isolates from a surgical ward at Hacettepe University Hospital. All MRSA isolates obtained from clinical specimens in this ward (14 MRSA isolates from wound cultures of 10 patients) were collected prospectively for 10 weeks. Surveillance cultures were taken from ward personnel (nose cultures from 4 physicians, 7 nurses, 1 secretary, 1 waiter), 2 surgical dressing containers and 1 nebulizer. MRSA was isolated from one of the surgical dressing containers, the nebulizer and nose cultures of 3 physicians, 3 nurses and the ward secretary. Arbitrarily primed polymerase chain reaction (AP-PCR) analysis showed that most MRSA isolates belonged to 2 major clones (pattern A, pattern B). Pattern A was the most frequent one and was present in 4 clinical isolates, surgical dressing container-1. Pattern B was identified in 3 clinical isolates and nose culture of physician-3. AP-PCR analysis revealed that this mini MRSA outbreak was caused by contamination of surgical dressing container with MRSA and nasal MRSA carriage in ward staff. AP-PCR seems to be a valuable typing method for analysis of nosocomial MRSA outbreaks because of its simplicity and rapidity. PMID- 10789554 TI - High prevalence of extended spectrum beta-lactamase production among Klebsiella pneumoniae strains isolated at a University Hospital in Turkey. AB - Beta-lactam susceptibility and beta-lactamase patterns of a random sample of 44 Klebsiella pneumoniae strains that had been isolated from nosocomial infections at Dokuz Eylul University Hospital in Izmir, were investigated. All strains were amoxycillin resistant but in the presence of clavulanic acid 26 became sensitive. Similarly 39 of the strains were resistant to ceftazidime and cefotaxime; clavulanic acid restored sensitivity to ceftazidime in 28 and to cefotaxime in 25 of these resistant strains. Extended spectrum beta-lactamase (ESBL) production was positive in 84% of the isolates as determined by the double disk synergy test. Isoelectric focusing revealed that each strain produced one to four beta lactamases, pI 7.6 enzymes being the most prevalent. Other enzymes with pIs of 8.4, 8.2, 5.4, 7.8 were also detected. Resistance to ceftazidime was transferred from 18 of the 44 isolates to the recipient Escherichia coli K-12 at 37 degrees C. The transconjugants were examined for their plasmid content and the plasmids were characterized by their size and resistance profile. Fourteen different restriction pattern groups were identified with Eco R1. The results indicate a high prevalence of ESBL production in nosocomial K. pneumoniae isolates in Izmir and have major implications concerning the clinical use of later generation cephalosporins. PMID- 10789555 TI - Antibiotic drug prescription in respiratory tract infections: a pharmacoepidemiological survey among general practitioners in a region of Italy. AB - Data concerning patients undergoing antibiotic treatment for upper (URTI) or lower (LRTI) respiratory tract infections were collected from 23 General Practitioners (GPs) in the Campania Region of Italy from November 15, 1997 to March 15, 1998. The objectives of the study were: a) to assess the occurrence of URTIs and LRTIs; b) to document the factors that influence GPs' choice of therapy; c) to correlate antibiotic choice with duration and outcome of treatment; d) to assess the incidence of unwanted effects. 2198 questionnaires were collected. Patients were +/-43.9 of age. URTIs were diagnosed in 65.4% and 34.6% LRTIs. The mean duration of antibiotic treatment was 4.5 days in URTIs and 5.6 days in LRTIs. The choice of antibiotic treatment was influenced by clinical assessment of infections (67.1%). The most commonly used antibiotic categories in URTIs were macrolides (39.3%), penicillins (27.4%) and cephalosporins (23.8%) whereas for LRTIs mainly cephalosporins (63.8%), penicillins (9.2%) and fluoroquinolones (7.4%) were used. Adverse events were experienced by 3.9% of patients. PMID- 10789556 TI - Effect of interleukin-2 on generation of natural killer cells: role of major histocompatibility complex class I in B6 and TAP-1-/- mice. AB - Long-term bone marrow cultures were used to investigate the effect of IL-2, a cytokine widely used in immunotherapy, on natural killer cell differentiation. Specifically, the role of MHC was evaluated by comparing normal B6 and class I deficient TAP-1-/- mice. The number of cells generated after a 13-day culture was the same in cell cultures from TAP-1-/- or B6 mice but the relative number of natural killer cells, identified as NK-1.1+CD3- cells by flow cytometry analysis, was increased in TAP-1-/- compared to B6 cultures (74.4% and 63.9%, respectively). Addition of an anti-class I mAb determined a strong inhibition of natural killer cell generation in B6 cultures, and its effect was specific since no effect was seen in TAP-1-/- cell cultures. TAP-1-/- natural killer cells or the few natural killer cells escaping the inhibitory effect of anti-class I mAb, were less cytotoxic than total B6 natural killer cells against target cell lines of different haplotype. PMID- 10789557 TI - Staurosporine increases carcinoembryonic antigen expression in a human colon cancer cell line. AB - Staurosporine (ST), a protein kinase C inhibitor, was found to produce antitumor effects against C22.20, a clonal subline derived from colon cancer HT-29 line, selected for low expression of carcinoembryonic antigen (CEA). However, as assessed by FACS analysis using propidium iodide, no apoptosis or cell cycle alteration was found on day 3 after treatment of C22.20 cells with ST (1-100nM). Exposure of cells to graded concentrations of the drug (i.e., from 1 to 25nM) resulted in a concentration-dependent increase in the percentage of CEA positive cells, as determined by flow cytometric analysis. However, when higher concentrations (i.e. 50nM - 100nM) of ST were used, the percentage of CEA positive cells declined compared to that detected in 25nM-treated tumor. Since these results were obtained in a clonal cell population, it is reasonable to hypothesize that induction rather than selection mechanism is involved in this phenomenon. The potential clinical interest of the present findings stems from the consideration that treatment with ST or its derivatives could improve sensitivity and efficacy of diagnostic and/or immunotherapeutic approaches based on CEA molecules. PMID- 10789558 TI - Cytogenetic toxicity of cisplatin in bone marrow cells of Swiss mice. AB - The inorganic platinum compound cisplatin (CP) in Oncoplatin, an anticancer drug, as the test chemical and cyclophosphamide (CY) in Endoxan, another anticancer drug, as the positive control, were tested for their cytogenetic toxicity in bone marrow cells of Swiss mice. The end points selected were mitotic metaphase chromosomal aberration and mitotic index study at 24-hour post-treatment and micronucleus test at 30-hour post-treatment after a single intraperitoneal exposure. The doses of the chemicals tested were CP 2, 3 and 5 mg/kg and CY 40 mg/kg b.w. of mice. Each of the doses of CP induced a significant number of chromosomal aberrations, mostly chromatid breaks and fragments and a significant number of micronuclei. The mitotic index study indicated CP as nonmitotoxic. The female mice were found more sensitive to each of the doses of CP than the males by showing more chromosomal aberrations, a higher number of micronuclei and comparatively less percentages of dividing cells. CP was thus found to be highly clastogenic in bone marrow cells of Swiss mice. PMID- 10789559 TI - Phase II study of weekly high dose fluorouracil in previously treated patients with metastatic colorectal cancer. AB - The purpose of this study was to evaluate the efficacy, toxicity, and safety of outpatient chemotherapy with weekly high-dose 5-fluorouracil (HD-5FU) in previously treated patients (pts) with metastatic colorectal cancer. Previously treated and failed pts with histologically confirmed, measurable metastatic colorectal cancer, performance status (WHO) 0-2 and adequate bone marrow function were eligible for treatment. Patients received 5FU (2.6 g/m2) as a 24-hour continuous infusion. Treatment was repeated weekly. A total of 202 cycles were given. Eighteen pts were enrolled. No pts achieved objective response. In 6 pts (33%), after 4 weeks of treatments, CEA level decreased 25% or more, and after 8 weekly treatments it rose again. Mild myelosuppression rarely occurred. Grade I nausea and vomiting occurred in 2 pts and Grade I diarrhea occurred in 2 pts. Mucositis was not observed. CONCLUSION: In our experience single agent, weekly, high dose 5-FU is well tolerated, but is ineffective as second-line treatment for metastatic colorectal cancer, and has only a marginal effect on CEA level. PMID- 10789560 TI - Comparative analysis of azithromycin and doxycycline efficacy in the treatment of female patients with acute urethral syndrome caused by Ureaplasma urealyticum. PMID- 10789561 TI - Candida famata fungemia in a cancer patient: case report. PMID- 10789562 TI - A look back. PMID- 10789564 TI - Comparison of two doses of aprotinin in patients receiving aspirin before coronary bypass surgery. AB - This study was designed to evaluate efficacy and tolerability of two different doses of aprotinin in patients receiving aspirin before undergoing coronary artery bypass grafting. Forty-two patients were randomized to receive either placebo (group I), or aprotinin in doses of 4,000,000 KIU (group II) or 6,000,000 KIU (group III). Drug efficacy was determined by measuring postoperative blood loss and transfusion of blood products. Both doses were effective in reducing blood loss and transfusion requirements. Blood loss through thoracotomy drainage was 450 +/- 224, 182 +/- 144, 142 +/- 98 ml, respectively, for control and treatment groups II and III (p = 0.0001). The numbers of patients with blood transfusions were seven (50%), two (17%) and two (17%) for group I and treatment groups II and III, respectively (p = 0.10). Tolerability was excellent and complications few and reversible. In conclusion, high and medium doses of aprotinin were well tolerated and reduced bleeding and transfusion requirements in patients submitted to coronary bypass surgery under the effects of aspirin. PMID- 10789563 TI - The role of nitric oxide synthase/nitric oxide in vascular smooth muscle control. AB - Vascular compliance is dependent on endogenous and exogenous sources of nitric oxide (NO). In a discussion of therapeutics and NO derived via nitric oxide synthase (NOS) enzymes, it is necessary to examine the pathways of each drug to provide the clinical perfusionist with a greater understanding of the role of NOS/NO in vascular function. Endothelial-derived NO is a contributor in the vasoregulation of vascular smooth muscle. Therapeutics seek to mimic the vasodilatory effects of the endogenous NO. The therapeutics included in this review are nitroglycerin, nitroprusside, amyl nitrite, and inhalation of NO. L Arginine supplementation provides additional substrate for the endogenous pathway that can augment NO production. NO is a small bioactive molecule involved in various biochemical pathways. Dysregulation of NO production can impair normal physiologic control of vascular compliance. Therefore, the purpose of this review is to provide the perfusionist with an understanding of the biochemical and pharmacological aspects of NOS/NO associated with vascular function. PMID- 10789565 TI - Impact of membrane oxygenators on pulsatile versus nonpulsatile perfusion in a neonatal model. AB - We investigated the effects of two new hollow-fiber membrane oxygenators, the Capiox SX10 and the Lilliput 901, on pulsatile versus nonpulsatile perfusion in an in vitro model designed to simulate a 3 kg infant. The experiments were divided into eight groups (six pulsatile and two nonpulsatile), according to the equipment and settings used. Each group included six tests. In all experiments, the pseudo-patient's mean arterial pressure was 40 mmHg, and the pump flow rate was 550 ml/min. During pulsatile cardiopulmonary bypass, the pump's base flow was set at 30%, and the pump rate was set at 80, 100, 120, 140, or 150 beats/min. The PUMP START and PUMP STOP timing points were adjusted to produce different pulse width settings. We were especially interested in evaluating the pre- and postoxygenator extracorporeal circuit pressure (ECP), the oxygenator pressure drop, and the precannula ECP. When used with a pulsatile roller pump, the Capiox produced a significantly lower preoxygenator ECP than the Lilliput (p < 0.001); moreover, the Capiox yielded a significantly lower oxygenator pressure drop (p < 0.001). During nonpulsatile perfusion, the Capiox again produced a lower preoxygenator ECP than the Lilliput (p < 0.001). These results suggest that the Capiox may be more suitable than the Lilliput when the pulsatile flow is employed, and pulsatile flow does not increase the ECP with either oxygenator. PMID- 10789566 TI - The effects of cardiopulmonary bypass and deep hypothermic circulatory arrest on blood viscoelasticity and cerebral blood flow in a neonatal piglet model. AB - The purpose of this study is to determine the effects of cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) on the viscoelasticity (viscosity and elasticity) of blood and global and regional cerebral blood flow (CBF) in a neonatal piglet model. After initiation of CPB, all animals (n = 3) were subjected to core cooling for 20 min to reduce the piglets' nasopharyngeal temperatures to 18 degrees C. This was followed by 60 min of DHCA, then 45 min of rewarming. During cooling and rewarming, the alpha-stat technique was used. Arterial blood samples were taken for viscoelasticity measurements and differently labeled microspheres were injected at pre-CPB, pre- and post-DHCA, 30 and 60 min after CPB for global and regional cerebral blood flow calculations. Viscosity and elasticity were measured at 2 Hz, 22 degrees C and at a strain of 0.2, 1, and 5 using a Vilastic-3 Viscoelasticity Analyzer. Elasticity of blood at a strain = 1 decreased to 32%, 83%, 57%, and 61% (p = 0.01, ANOVA) while the viscosity diminished 8.4%, 38%, 22%, 26% compared to the baseline values (p = 0.01, ANOVA) at pre-DHCA, post-DHCA, 30 and 60 min after CPB, respectively. The viscoelasticity of blood at a strain of 0.2 and 5 also had similar statistically significant drops (p < 0.05). Global and regional cerebral blood flow were also decreased 30%, 66%, 64% and 63% at the same experimental stages (p < 0.05, ANOVA). CPB procedure with 60 min of DHCA significantly alters the blood viscoelasticity, global and regional cerebral blood flow. These large changes in viscoelasticity may have a significant impact on organ blood flow, particularly in the brain. PMID- 10789567 TI - Size makes a difference: use of a low-prime cardiopulmonary bypass circuit and autologous priming in small adults. AB - Low hematocrit (Hct < 20) during cardiopulmonary bypass (CPB) is associated with higher mortality and other adverse outcomes. More frequently, low Hct is encountered in patients with small body size and women patients. This prompted us to take an aggressive approach in our care of these patients, involving a strategy for predicting patients at risk of low Hct, with the aid of an electronic worksheet that accurately predicts CPB Hct, and two prevention strategies: use of a low-prime CPB circuit (LP) for all adult patients with a body surface area (BSA) < 1.7 m(2) and use of autologous circuit priming (AP), in addition to the low-circuit volume in some patients. The two cohorts of patients in whom these techniques were employed were compared to a group matched for body size where our standard adult circuit (STD) was used. There were 233 patients in the standard group, 139 in the LP group, and 68 in the LP/AP group. The CPB circuit prime volume was 1,710 ml for the STD group and 1,110 ml for the LP group. Use of autologous priming techniques further reduced the prime volume by 545 +/- 139 ml. The incidence of low Hct (<20%) during CPB was thus reduced from 70% to 15% (p = 0.001) when using both techniques together without increasing red blood cell (RBC) transfusions. These changes in perfusion management resulted in a reduction in the incidence of renal complications (STD = 9.4%, LP = 6.5% (ns) and LP/AP = 0%, PMID- 10789568 TI - Point-of-care testing: inspection preparedness. AB - Point-of-care testing (POCT) in the operating room has changed dramatically since the implementation of the Clinical Laboratory Improvement Amendments (CLIA '88), which became effective in September 1992. With the implementation of CLIA '88, the Health Care Financing Administration (HCFA) mandated that human specimen testing 'for the purpose of diagnosis, prevention, or treatment of any disease or impairment of, or the assessment of the health of human beings', must be performed by a certified laboratory or testing site. To attain and maintain accreditation, the need for more stringent and comprehensive documentation has become imperative. The Joint Commission for the Accreditation of Hospitals (JCAHO), the College of American Pathologists (CAPS), HCFA, and state regulatory agencies require data such as staff credentialling, staff training/competency, procedure manuals, quality control logs, quality assurance/corrective action plans, correlation studies, proficiency testing results, and equipment maintenance logs to assure specimens are analyzed in a reliable manner by competent personnel so as not to jeopardize the safety and well being of the patient. Developing a comprehensive, ongoing survey readiness plan that includes a pre-survey checklist of all the documentation required and having this documentation in order and up to date well in advance of the survey will greatly enhance the probability of a successful survey conducted by the various regulatory agencies. PMID- 10789569 TI - The effects of modified hemofiltration on inflammatory mediators and cardiac performance in coronary artery bypass grafting. AB - Cardiopulmonary bypass increases the blood levels of various immune mediators, thereby leading to a systemic inflammatory response syndrome, e.g. sepsis, with some hemodynamic alterations, such as vasodilatation, tachycardia, and a decrease in systemic vascular resistance. Perioperative hemofiltration is one of the treatment modalities proposed to prevent this syndrome. Modified hemofiltration has been introduced recently by investigators who recommend that the former standard techniques are ineffective in eliminating the inflammatory mediators. The purpose of this study was to determine the effects of the modified technique on these mediators and on hemodynamic parameters. Forty patients undergoing coronary artery bypass grafting were randomized into equal control and hemofiltered groups. The hemodynamic parameters, as well as blood samples, were taken before and after hemofiltration to assess blood concentrations of interleukin-6, interleukin-8 and neopterin. The hemodynamic parameters and immune mediator levels did not differ between the two groups during the course of the study, except in the immediate postoperative periods, where cardiac output, cardiac index, and systemic vascular resistance values were significantly greater in the hemofiltered group while there were no differences in the immune mediators. The results of our study suggest that the effects of modified hemofiltration on immune mediators are still debatable. The improvement found in cardiac performance could be attributed to the prevention of hemodilution and hypervolemia. PMID- 10789570 TI - Carbon dioxide field flooding techniques for open heart surgery: monitoring and minimizing potential adverse effects. AB - The use of carbon dioxide for displacement of air in cardiac surgery can have potential adverse effects on blood gas strategies. Presented is a method of monitoring carbon dioxide in the cardiopulmonary bypass circuit and limiting the potential for severe hypercarbia during cardiopulmonary bypass. PMID- 10789571 TI - Reoperation for a type I aortic dissection: case report. AB - Surgery for the repair of a type I aortic dissection presents several difficulties for the surgeon and the perfusionist. One must safely support the patient, while at the same time provide the surgeon with a bloodless field in which to operate. Often, this requires cessation of the circulation for varying amounts of time. Deep hypothermia allows for an extension of the arrest period, while other techniques-- retrograde cerebral perfusion and antegrade cerebral perfusion--provide an additional degree of cerebral protection. Recently, we utilized these techniques concurrently on a 43-year-old female who presented for a reoperation for a type I aortic dissection. Combining these techniques allowed us to adequately support the patient during an anticipated lengthy period of circulatory arrest and insured a successful operation without any adverse cerebral or other organ dysfunction. PMID- 10789572 TI - Pre-operative coagulopathy management of a neonate with complex congenital heart disease: a case study. AB - Severe coagulation defects often develop in neonates undergoing cardiac surgery, both as a result of the surgical intervention, and as pre-existing defects in the hemostatic mechanisms. The following case report describes a newborn patient with complex congenital heart disease and respiratory failure whose pre-operative coagulopathy was aggressively managed prior to surgical correction. A 5-day-old, 2.5 kg child presented with interrupted aortic arch, ventricular septal defect, atrial septal defect, and patent ductus arteriosus. On admission, he was in respiratory arrest suffering from profound acidemia. In addition, the child was hypothermic (30.1 degrees C), septic (Streptococcus viridans), and coagulopathic (disseminated intravascular coagulation-DIC). The patient was immediately intubated and initial coagulation assessment revealed the following: prothrombin time (PT) 48.9 s (international normalized ratio (INR) 15.7), activated partial thromboplastin time (aPTT) >106 s, platelet count 30,000 mm(3), fibrinogen 15 mg dL(-1) and antithrombin III (AT-III) 10%. Before cardiac surgery could be performed, the patient's DIC was corrected with the administration of cryoprecipitate (15 ml), fresh frozen plasma (300 ml), and platelets (195 ml). In spite of the large transfusion of fresh frozen plasma, the AT-III activity, measured as a percentage, remained depressed at 33. Initial thromboelastographic (TEG) determination revealed an index of +2.02, and following 100 IU administration of an AT-III concentrate, declined to -2.32. Sequential TEG profiles were performed over several days, with the results used to guide both transfusion and medical therapy. The congenital heart defect correction was subsequently performed with satisfactory initial results, but the patient developed a fungal infection and expired on the 16th post-operative day. The present case describes techniques of coagulation management for a newborn with both a severe hemostatic defect and congenital heart disease. PMID- 10789573 TI - Extracorporeal support in an adult with severe carbon monoxide poisoning and shock following smoke inhalation: a case report. AB - The objective of this study was to discuss the case of a patient with severe smoke inhalation-related respiratory failure treated with extracorporeal support. The study was set in a 12-bed multi-trauma intensive care unit at a level one trauma center and hyperbaric medicine center. The patient under investigation had carbon monoxide poisoning, and developed acute respiratory distress syndrome and cardiovascular collapse following smoke inhalation. Rapid initiation of extracorporeal support, extreme inverse-ratio ventilation and intermittent prone positioning therapy were carried out. Admission and serial carboxyhemoglobin levels, blood gases, and computerized tomography of the chest were obtained. The patient developed severe hypoxia and progressed to cardiovascular collapse resistant to resuscitation and vasoactive infusions. Veno-venous extracorporeal support was initiated. Cardiovascular parameters of blood pressure, cardiac output, and oxygen delivery were maximized; oxygenation and ventilation were supported via the extracorporeal circuit. Airway pressure release ventilation and intermittent prone positioning therapy were instituted. Following 7 days of extracorporeal support, the patient was decannulated and subsequently discharged to a transitional care facility,neurologically intact. Smoke inhalation and carbon monoxide poisoning may lead to life-threatening hypoxemia associated with resultant cardiovascular instability. When oxygenation and ventilation cannot be achieved via maximal ventilatory management, extracorporeal support may prevent death if initiated rapidly. PMID- 10789574 TI - Is it possible to resterilize disposable laparoscopy trocars in a hospital setting? AB - Nosocomial infections associated with interventional procedures have been attributed to improper decontamination of instruments. Disinfection of solid laparoscopic instruments, such as telescopes, by 2% glutaraldehyde and ethylene oxide was shown to be effective in preventing infection transmission. However, instrument design in more complex surgical instruments may hamper the quality of disinfection. The aim of this study is to investigate the safety of hospital disinfection of disposable laparoscopic instruments with a relatively more complex design. A total of 40 laparoscopic trocars were divided into two equal groups: group 1 was contaminated with bacteria and yeast, and group 2 was contaminated with the hepatitis B virus. Each group was then divided to two equal subgroups. After disinfecting subgroup A with 2% glutaraldehyde and B with ethylene oxide, samples were obtained for bacterial cultures and for virus detection using polymerase chain reaction (PCR). Bacterial and yeast cultures were positive in three instruments in group 1A and in two instruments in group 1B. Tests results for the hepatitis B virus were negative in group 2A, but positive in group 2B. Results of this study indicate that disinfection for multiple use of disposable laparoscopic instruments with a relatively complex structure is not effective and may result in nosocomial disease transmission by bacteria, fungi, and viruses. PMID- 10789575 TI - Laparoscopic colectomy for Dukes A colon cancer. AB - The use of laparoscopic surgery to treat colon cancer has been well studied; however, the specific use of laparoscopic colectomy for Dukes A colon cancer has not been evaluated. The data of laparoscopic colectomy were compared with those of conventional open colectomy, and the surgical results of patients who underwent surgery for Dukes A colon cancer were evaluated. Between November 1993 and October 1997, 20 patients underwent laparoscopic colectomy for Dukes A colon cancer. Operation time, blood loss, first passage of flatus, day of resumption of oral intake, length of hospital stay after surgery, and number of dissected lymph nodes were compared between 20 patients who underwent laparoscopic colectomy and 23 patients who underwent conventional open colectomy for Dukes A colon cancer. In patients with laparoscopic colectomy, when compared with those with conventional open colectomy, mean blood loss was less (103 g vs. 318 g), flatus returned more quickly (3.5 days vs. 4.2 days), oral intake resumed earlier (3.7 days vs. 4.7 days), and postoperative hospital stay was shorter (16.4 days vs. 24.6 days). The mean number of dissected lymph nodes was not different between the two groups (9.2 vs. 9.2 for D2 dissection). No patient had port-site metastasis or recurrence during a follow-up period from 13 to 60 months (median, 38 months). Review of the literature and the authors' findings indicated that none of the 142 reported patients had port-site metastasis after laparoscopic colectomy for Dukes A colon cancer. The results indicate that laparoscopic colectomy is safe and useful when applied to patients with Dukes A colon cancer and performed carefully by trained surgeons. PMID- 10789576 TI - Video-mediastinoscopy in management of patients with lung cancer: a preliminary study. AB - Cervical mediastinoscopy is widely employed for biopsy of mediastinal lymph nodes and staging of lung cancer. The application of video-assisted technology to mediastinoscopy in a series of patients with lung cancer has not been reported. Preliminary experience with the use of video-mediastinoscopy in diagnosis and staging of lung cancer is presented. Fifteen patients with lung cancer were studied. Results of previous computed tomography scans had shown the presence of enlarged mediastinal lymph nodes in the retrovascular plane in all of the cases. Video-mediastinoscopy was performed under general anesthesia using a specifically designed rigid scope connected to a mono-charged-coupled device video camera (model INH 002756; Karl Storz-Endoskope, Tuttlingen, Germany). Neither fatalities nor major complications related to the procedure were observed. In all cases, video-mediastinoscopy proved useful for diagnosis or staging of lung cancer, therefore contributing to clinical decision making. The optimal visualization of mediastinal structure and the possibility for the surgeon to operate with both hands are appreciable characteristics of this technique. PMID- 10789577 TI - Descending aorta-to-femoral artery bypass: preliminary experience with a thoracoscopic technique. AB - Descending aorta-to-femoral artery bypass is a durable procedure with excellent long-term patency rates. The operation is usually performed using a lateral thoracotomy combined with retroperitoneal tunneling of the graft. Assuming that a smaller incision would reduce the operative trauma, minimally invasive video endoscopic techniques were used to perform a thoracoscopic bypass operation. In all patients, exposure of the descending aorta was obtained using thoracoscopy. This was accomplished with a maximum of four ports. For placing the conventional side-biting aortic clamp, a 3- to 4-cm incision was necessary. Using this incision as an access port, the proximal anastomosis was sutured endoscopically. Retroperitoneal tunneling was performed using laparoscopically guided balloon dissection. Eleven consecutive patients underwent surgery. In two patients, conversion to a standard thoracotomy was necessary because of extensive intrathoracic adhesions. There were no major complications, except for one hematoma. After a mean follow-up of 9.5 months, all bypass grafts were still patent. Patients could be discharged after a mean postprocedural hospital stay of 7.4 days (range, 5-12). Thoracoscopic descending aorta-to-femoral artery bypass is an operation with excellent patency rates; however, it is a novel technique that still requires further technical improvements. Clinical studies are needed to prove the safety and efficacy of this minimally invasive technique. PMID- 10789578 TI - The mini-laparoscopic intragastric resection of a gastroesophageal stromal tumor: a novel approach. AB - The laparoscopic wedge resection of gastric stromal tumors has been shown to be safe and effective. The removal of these tumors at the gastroesophageal junction is not amenable to a typical local resection because of anatomic inaccessibility and potential luminal restriction after resection. Also, an esophagogastrectomy is not tenable in the setting of benign disease. The recent advances in miniaturized laparoscopic instrumentation (2-mm diameter) have expanded the laparoscopic options, including intraluminal resection. The first intragastric mini-laparoscopic resection of the gastric stromal tumor at the gastroesophageal junction is reported. PMID- 10789579 TI - Port site recurrence of gallbladder cancer after laparoscopic surgery: two case reports of long-term survival. AB - More than 100 patients with port site recurrence after laparoscopic procedures have been reported, and in most cases recurrence has had a fatal outcome. Two patients who survived port site recurrence of unexpected gallbladder cancer after laparoscopic cholecystectomy are reported. Abdominal wall excision was performed in one patient, and thermoradiotherapy was performed in the other. Both patients have remained free of disease during long-term follow-up (82 and 45 months). PMID- 10789580 TI - Uncommon left hepatic duct injury during laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy is currently the gold standard in the treatment of symptomatic gallstones but has been shown to have a higher incidence of biliary tree lesions (0.3-1%) compared with reported traditional open approaches. Loss of three-dimensional view and of depth perception is the main limit of the laparoscopic approach, especially if particular risk factors are associated (e.g., postinflammatory fibrosis, anatomic variations). Moreover, inadequate training may justify the increase of biliary tract lesions. The authors describe a unique case of left hepatic duct clipped without section of the duct itself during an otherwise "easy" operation. At the reintervention, because of the favorable local condition, a reconstruction was possible after a small duct resection with a ductal-hepatic anastomosis over a T-tube. This was removed after 8 months because of the good patency of the biliary tree and the absence of cholestasys. A long-term follow-up is mandatory for a complete functional evaluation. PMID- 10789581 TI - Retroperitoneal abscess after retained stones during laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy is associated with a significant risk of gallbladder perforations with bile and stone spillage. The retrieval of dropped stones is sometimes impossible, and intraperitoneally retained stones can be the source of serious complications, such as inflammatory masses or abscesses. The authors describe a patient in whom a large retroperitoneal abscess developed as a result of missed stone fragments during cholecystectomy. Although several cases of intraperitoneal abscess have been reported in the literature, retroperitoneal collection is very uncommon. Crushed and infected stones seem to be the essential prerequisite for abscess formation. Therefore, every attempt should be made to avoid stone spillage and intra-abdominally retained stones. Conversion to open surgery has to be considered in the presence of adverse factors, such as primary acute cholecystitis or cholecystitis induced by previous sphincterotomy, or bilirubinate stones believed to be infected. Abdominal abscess treatment requires removal of missed stones by surgical incision or new laparoscopic procedure because simple percutaneous drainage is usually unsuccessful. PMID- 10789582 TI - Acute intrathoracic incarceration of the stomach after laparoscopic Nissen fundoplication. AB - A rare case of paraesophageal hernia with complete intrathoracic incarceration of the stomach after laparoscopic Nissen fundoplication is described. An 85-year-old woman who had undergone laparoscopic Nissen fundoplication for gastroesophageal reflux disease presented 14 months later with nausea and vomiting. Esophagogastroendoscopy showed obstruction of the esophagogastric junction and gastric mucosal necrosis. Emergency laparotomy showed the stomach to be entirely strangulated into the thorax, with areas of necrosis. Gastrotomy was followed by resection of the necrotic anterior wall of the stomach, closure of the hiatus, and suturing of the stomach to the diaphragm. Appropriate closure of crura and anchoring suture between the stomach and diaphragm are helpful to prevent recurrent hernia after laparoscopic Nissen fundoplication. PMID- 10789583 TI - Laparoscopic treatment for biliary ascariasis. AB - Biliary ascariasis is one of the most common types of ascaris infections. The current treatments are helminthic drug therapy, endoscopic extraction, and surgical extraction. A case of biliary ascariasis and cholecystocholedocholithiasis was successfully treated by laparoscopic extraction of the living worm and biliary stones. This procedure was found to be very effective for biliary ascariasis with biliary stones, and it holds promise for similar cases in the future. PMID- 10789584 TI - Technique for laparoscopic partial splenectomy. AB - Splenic preservation and conservative management are now accepted norms when dealing with splenic pathologic conditions. This case report describes the technique of laparoscopic partial splenectomy for an undiagnosed splenic lesion. A thorough understanding of splenic anatomy permits laparoscopic partial splenectomy with the resultant benefits, including a decreased risk of postsplenectomy sepsis, short hospital stay, and superior cosmesis. The success and relative ease of performing this procedure will pave the way for its future use in other selective cases involving splenic pathology. PMID- 10789585 TI - Agenesis of the vermiform appendix. AB - Agenesis of the vermiform appendix is very rare. The incidence is estimated to be one in 100,000 laparotomies for suspected appendicitis. Several criteria have to be met before the investigator can conclude that the appendix is congenitally absent. This case is reported to bring this entity to the attention of surgeons who may encounter a similar situation during celioscopy. A 29-year-old patient was admitted through the emergency room with the chief complaint of abdominal pain. Acute appendicitis was suspected, and he was accordingly prepared for celioscopy. This report presents a patient with vermiform appendix agenesis diagnosed at celioscopy with concomitant mesenteric lymphadenitis. Agenesis of the vermiform appendix is very rare, and the diagnosis should not be made unless the ileocecal and retrocecal area are thoroughly explored. PMID- 10789586 TI - A measurement of the ability to drive after different types of inguinal hernia repair. PMID- 10789587 TI - New calmodulin antagonists inhibit in vitro growth of human breast cancer cell lines independent of their estrogen receptor status. AB - Calmodulin plays a key role in the regulation of cell proliferation and calmodulin antagonists may offer a new therapeutic approach in the treatment of breast cancer. Three new specific calmodulin antagonists with improved potency were synthesized and screened on human breast cancer cell lines known to be estrogen receptor (ER)-positive or -negative. These calmodulin antagonists significantly inhibited cell growth as measured by the MTT proliferation assay (p<0.001). Their IC50 values were in the low micromolar range against both ER positive and -negative variants of the MCF-7 cell line. Two other breast cancer cell lines (ER-positive T-47D and ER-negative MDA-MB-231) were also inhibited by these calmodulin antagonists with IC50 values in a similar range. The level of inhibition was independent of any stimulation of cell growth by estradiol. Calmodulin antagonists effectively reduced cell growth of both ER-positive and negative human breast cancer cell lines in vitro. Calmodulin antagonists represent a novel therapeutic approach requiring further investigation. PMID- 10789589 TI - The effects of granulocyte colony stimulating factor on chemiluminescence and lipid peroxidation of blood platelets treated with cisplatin. AB - The effects of granulocyte colony stimulation factor (G-CSF) at concentrations of 0.08, 0.8 and 8 microg/ml on reactive oxygen species (ROS) generation and lipid peroxidation induced by cisplatin in pig blood platelets were investigated. The level of reactive oxygen species (O2, H2O2, singlet oxygen and organic radicals) generated in platelets was measured by the chemiluminescence method. Lipid peroxidation was determined by the thiobarbituric acid technique and was expressed as thiobarbituric acid reactive substances. G-CSF at the concentration of 0.08 microg/ml had a strong inhibitory effect (about 60% inhibition) on the production of ROS in the isolated pig platelets. This cytokine also significantly reduced lipid peroxidation in control platelets and platelets treated with cisplatin (p<0.05). In the presence of G-CSF in the incubation medium (0.8 microg/ml) cisplatin-induced generation of ROS was also reduced (p<0.05). This study demonstrates that G-CSF has a protective effect against the oxidative stress in blood platelets caused by cisplatin. PMID- 10789588 TI - Angiostatic effects of suramin analogs in vitro. AB - Suramin analogs are polyanionic naphthylureas structurally related to suramin, an antitumor agent with a narrow therapeutic window. The angiostatic activities of suramin and 16 suramin analogs were investigated using an easily quantifiable in vitro angiogenesis system. In addition, the antiproliferative activities of the analogs were studied in four different human tumor cell lines and in porcine aortic endothelial cells. The suramin analogs encompassed two main structural variations, i.e. their molecular size, and the number and substitution pattern of the sulfonate groups. Some suramin analogs with a reduced number of sulfonate groups (NF062, NF289 and NF326) showed significant dose-dependent angiostatic and also antiproliferative activities. The disulfonate NF062 was superior to suramin in inhibiting HT29 and T47D tumor cells while demonstrating a similar angiostatic potential as suramin. Therefore, the sulfonate groups in the para position of the amino groups of the naphthyl residues of suramin seem to be of special importance. The very small disulfonates (NF108, NF109, NF499, NF500 and NF241) and the asymmetric compound NF520, one half of the suramin molecule, are inactive. Therefore, a minimal molecule size seems to be essential for the biological activity. Suramin is a rather rigid molecule. The highly flexible analogs (NF527, NF528 and NF529) are inactive. This indicates that the molecular rigidity is important for the biological activity. PMID- 10789590 TI - Determination of the maximal carcinoma/normal skin ratio after HpD or m-THPC administration in Hairless mice (SKH-1) by fluorescence spectroscopy. AB - The two major steps in our study on the treatment of skin carcinomas by photochemotherapy (PCT) were the development of a skin tumor model in Hairless mice by a chemical carcinogenesis and the use of fluorescence spectroscopy, a semi-quantitative and non-invasive method, in order to determine the time after i.p. injection of photosensitizer when the tumor/normal skin ratio was the highest. A three-step carcinogenesis protocol provided mice bearing carcinomas and these were used to determine the tumor/normal skin ratios of two photosensitizers by fluorescence spectroscopy. Hematoporphyrin derivative (HpD) (5 mg/kg body weight) and m-tetra(hydroxyphenyl) chlorine (m-THPC) (0.3 mg/kg body weight) were injected i.p., and fluorescence was measured at 1, 4, 8, 12, 24, 48, 72 and 96 h after injection. The best carcinoma/normal skin ratio would be 3.2+/-1.4 for HpD and 2.7+/-2.1 for m-THPC, respectively. The delays required to reach these ratios were 72 h for HpD and 24 h for m-THPC. These results have to be considered with caution due to the high SEs and they must be confirmed by organic extraction. Photodynamic therapy with the same doses of HpD and m-THPC used in this pharmacokinetic study has to be carried out in order to compare the toxicities of the two photosensitizers and to determine which one is the best for this type of tumor. PMID- 10789591 TI - In vitro sensitivity of normal human mesothelial and malignant mesothelioma cell lines to four new chemotherapeutic agents. AB - In this study, we used four human mesothelioma cell lines (M14K, M24K, M25K and M38K), one transformed human mesothelial cell line (MeT-5A) and one primary mesothelial culture (UPL) to test for in vitro sensitivity to docetaxel, paclitaxel, SN-38 [an active metabolite of irinotecan (CPT-11)] and gemcitabine, as single agents. Subconfluent cell cultures were treated with 2x10(-9), 5x10( 9), 10(-8), 2x10(-8) and 5x10(-8) M concentrations of each drug for 48 h. The sensitivity was measured in terms of cell viability using the Trypan blue exclusion method. All four drugs were potent inhibitors of mesothelioma cell growth, but cell lines from different patients diverged in their sensitivity to the individual agents. In most cases docetaxel, paclitaxel and SN-38 were more potent killers of mesothelioma cells than gemcitabine. The induction of DNA damage was investigated using the Comet assay; cells from two cell lines (M14K and M25K) were treated with subtoxic 10(-8) M concentrations of each drug for 4, 24 and 48 h. Each of the agents caused a slight increase in DNA single-strand breaks at a concentration of 10(-8) M. PMID- 10789592 TI - Interferon-alpha and 13-cis-retinoic acid as maintenance therapy after high-dose combination chemotherapy with growth factor support for small cell lung cancer--a feasibility study. AB - This randomized phase II multi-center study was designed to determine the time to progression, duration of response and the feasibility of an intensified maintenance regime consisting of a combination of interferon (IFN)-alpha and retinoic acid after high-dose combination chemotherapy and radiotherapy in patients with small cell lung cancer. The patients received four courses of combination chemotherapy consisting of ifosfamide, carboplatin and etoposide, with higher doses of ifosfamide and carboplatin given in the first course, with routine growth factor support. Responding patients were then randomly assigned to one of three maintenance therapy arms. All patients with limited disease (LD) were given thoracic radiotherapy before maintenance therapy and those who had also achieved a complete response (CR) or minimal residual disease (MRD) received prophylactic cranial irradiation. In Arm 1 patients received IFN-alpha-2a, 6 MIU s.c. TIW for 4 weeks, followed by 3 MIU s.c. TIW, and 13-cis-retinoic acid 1 mg/kg/day p.o. BID daily. In Arm 2 patients received trophosphamide 100-150 mg/day p.o. BID. No maintenance treatment was given in Arm 3, the control group. Maintenance therapy was continued for 1 year. Eighty-five patients were treated according to the protocol. Twenty-one patients achieved CR, four achieved MRD and forty-two achieved partial responses to chemotherapy and radiotherapy. Sixty patients (71%) were randomly assigned for maintenance treatment. Median survival was 17.1 months in the IFN-alpha-retinoic acid arm, 12.4 months in the trophosphamide arm and 13.5 months in the control arm. One-year survival rates were 82, 56 and 55%, respectively. Duration of response was 6.5, 5.5 and 4.7 months, respectively. Time to progression was 8.6, 8.0 and 6.8 months, respectively The differences were not statistically significant. The IFN-alpha retinoic acid maintenance treatment was well tolerated. Patients who received IFN alpha-retinoid maintenance therapy lived longer after the onset of progressive disease. The treatment regime was effective, feasible and well tolerated. PMID- 10789593 TI - Multicenter study of the impact of prescription guidelines on the use of colony stimulating factors. AB - The aim of this work was to assess the impact of circulating guidelines for correct prescription practices of colony stimulating factors (CSF). Two hospital groups were compared, a 'guidelines' group (seven teaching hospitals) that circulated the guidelines and a control group (eight teaching hospitals) that did not. In addition, two periods were compared before and after distribution of the guidelines: from 17 February to 2 March 1996 and from 17 February to 2 March 1997. The assessment involved compliance with the guidelines for the following parameters: indications, dose regimen, time to start of CSF therapy and duration of CSF therapy between the control and guideline groups and also between the two periods. The population included 404 patients analyzed (209 in 1996 and 195 in 1997) for the indication of post-chemotherapy neutropenia. Total compliance in the first period (all four items) was 44.2% in the control group and 50.8% in the guideline group (nonsignificant), and during the second period was 31.9 and 59.6% in the two groups (p<0.001). During the first period, the differences in compliance with the guidelines for indication, dose regimen, time to start of treatment and duration between the groups were not significant. In the second period, this difference became significant and in favor of the guideline group for dose regimen (p = 0.009) and treatment duration (p = 0.02). The results of this study show the need to continuously define prescription reference systems according to available data, and to circulate them widely to improve the quality of health care and to control expenses. PMID- 10789594 TI - Monthly gemcitabine (days 1, 8 and 15) plus cisplatin (days 1-3) in advanced non small cell lung cancer: a phase II study. AB - On the basis of the reported efficacy of gemcitabine plus cisplatin in patients with non-small cell lung cancer (NSCLC), this combination has been selected to be given as our firstline service regimen for advanced or metastatic disease. Patients recruitment was almost unlimited: no exclusion criteria were made, except for disease-related Karnofsky's performance status below 50%, the presence of central nervous system or spinal involvement by uncontrolled metastases, or creatinine clearance below 50 ml/min. Cisplatin 30 mg/m2/day on days 1-3 and gemcitabine 1250 mg/m2/day on days 1, 8 and 15 every 4 weeks were given on an outpatient schedule to consecutive patients with locally advanced or metastatic NSCLC. Forty-three successive NSCLC patients with histologically or cytologically proven disease were treated. Adenocarcinoma was diagnosed in 35% of cases, squamous cell carcinoma in 60% and broncho-alveolar type in 5%. Smoking was mentioned by 63% of the patients. Numerous medical problems were recorded in 75% of the patients. Stage IIIB was observed in 10 of 43 patients, while metastatic disease was found in the rest. All the patients, except for two, were symptomatic. Two patients achieved complete response (5%) and 16 achieved partial response (37%), yielding an overall objective response rate of 42%. Minimal response was observed in seven patients (16%) and disease stabilization in 7%. Adding the objective response rate to the minimal response and stabilization rates, the disease-control (progression-free) rate reaches 65%. The time to progression ranged from 0 to 69 weeks in all the patients. The overall survival of the group ranged from 4 to 98 weeks, with a median of 45 weeks. Clinical benefit response was observed mainly in patients who also achieved an objective response. We conclude that outpatient cisplatin plus gemcitabine combination is feasible, efficacious and justified in patients with advanced or metastatic NSCLC. PMID- 10789595 TI - Doxil (Caelyx): an exploratory study with pharmacokinetics in patients with hormone-refractory prostate cancer. AB - Doxil, a doxorubicin formulation of polyethylene glycol-coated liposomes, has anti-tumor activity against Kaposi's sarcoma and other solid tumors with mild myelosuppression, minimal hair loss and a low risk of cardiotoxicity. Non liposomal doxorubicin has modest activity in hormone-refractory prostate cancer (HRPC) with considerable toxicity. A pilot study of Doxil was conducted in 15 patients with HRPC. Doxil was administered i.v. using two regimes of equal dose intensity, either 45 mg/m2 every 3 weeks or 60 mg/m2 every 4 weeks. Plasma levels of doxorubicin were analyzed in 10 patients. The most common side effect was stomatitis with a higher incidence at the 60 mg/m2 dose level. In contrast, hand foot syndrome was more frequent and severe in patients treated with the 3 week schedule of 45 mg/m2. Three patients responded to treatment (based on objective response in one patient and reduction of PSA level greater than 50% in the other two) and two patients had stable disease, all of them receiving 60 mg/m2. Pharmacokinetic analysis shows a proportional increase of plasma drug levels with dose and the characteristic long circulation time of Doxil with half-lives in the range of 3 days, somewhat longer than previously reported. In conclusion, Doxil at 60 mg/m2 every 4 weeks appears to be active against HRPC, but severe mucocutaneous toxicities prevented further investigation of this regime. PMID- 10789596 TI - A comparative pharmacokinetic study of doxorubicin and 4'-epi-doxorubicin in children with acute lymphocytic leukemia using a limited sampling procedure. AB - Antraquinone glycosides are an important class of antineoplastic drugs, frequently used for treatment of a variety of malignancies in children. Doxorubicin (Dox) is the most frequently used drug within this class of antineoplastics. 4'-epi-doxorubicin (Epi), a Dox isomer, was developed with the aim of reducing risks for fatal heart toxicity observed with Dox. The aim of the present study was to investigate the pharmacokinetics of Dox and Epi in children with acute lymphocytic leukemia. In total 31 patients (13 females and 18 males; median age 5.4 years; range 0.73-15.3 years) were studied using a simplified sampling procedure. The pharmacokinetic differences of the two drugs were established by their simultaneous administration. The plasma pharmacokinetics of neither Dox nor Epi correlated with the age of the patients. There were no gender differences in dose-normalized maximum concentrations of neither Dox nor of Epi. The inter-patient variation of the dose-normalized maximum concentrations of Dox and Epi is larger among females than among males. The Cmax ratio Dox/Epi was 1.39+/-0.19 (mean +/- SD). The pharmacokinetic differences of Dox and Epi in children, although less pronounced than in adults, are still of a magnitude that might be of clinical importance. PMID- 10789597 TI - Efficacy and safety of oral granisetron versus i.v. granisetron in patients undergoing peripheral blood progenitor cell and bone marrow transplantation. AB - This randomized, controlled, double-blind pilot study assessed the efficacy and safety of oral versus i.v. granisetron, both in combination with non-5-HT3 antiemetics, in preventing emesis caused by high-dose chemotherapy. Fifty-one patients who underwent peripheral blood progenitor cell transplantation (PBPCT) or bone marrow transplantation (BMT) were evaluated. Efficacy was assessed by the number of emetic episodes during the worst 24 h period. A complete response (CR) was defined as no vomiting, partial response (PR) as less than three emetic episodes and failure as three or more emetic episodes. Patients who received oral granisetron experienced significantly (p<0.0008) fewer emetic episodes than those who received i.v. granisetron; however, the number of emetic episodes over the worst 24 h was similar between the oral and i.v. granisetron groups (13 and 15, respectively), as were the overall response rates (CR+PR, 54.5 and 41.4%, respectively). Both dosage forms were well tolerated. Based on these findings, further comparative studies of oral granisetron are warranted in patients undergoing PBPCT or BMT. PMID- 10789598 TI - Update on the management of hyperthyroidism and hypothyroidism. AB - Clinical aspects, laboratory investigation, and treatment of hyperthyroidism and hypothyroidism are reviewed in light of recent information. Special circumstances, such as hyperthyroidism during pregnancy, Graves ophthalmopathy, iodine-induced hyperthyroidism, and subclinical hypothyroidism, are also considered. PMID- 10789599 TI - Hospital readmissions as a measure of quality of health care: advantages and limitations. AB - We reviewed the recent literature on hospital readmissions and found that most of them are believed to be caused by patient frailty and progression of chronic disease. However, from 9% to 48% of all readmissions have been judged to be preventable because they were associated with indicators of substandard care during the index hospitalization, such as poor resolution of the main problem, unstable therapy at discharge, and inadequate postdischarge care. Furthermore, randomized prospective trials have shown that 12% to 75% of all readmissions can be prevented by patient education, predischarge assessment, and domiciliary aftercare. We conclude that most readmissions seem to be caused by unmodifiable causes, and that, pending an agreed-on method to adjust for confounders, global readmission rates are not a useful indicator of quality of care. However, high readmission rates of patients with defined conditions, such as diabetes and bronchial asthma, may identify quality-of-care problems. A focus on the specific needs of such patients may lead to the creation of more responsive health care systems for the chronically ill. PMID- 10789600 TI - Pulse pressure not mean pressure determines cardiovascular risk in older hypertensive patients. AB - BACKGROUND: Current guidelines for the management of hypertension rest almost completely on the measurement of systolic and diastolic blood pressure. However, the arterial blood pressure wave is more correctly described as consisting of a pulsatile (pulse pressure) and a steady (mean pressure) component. OBJECTIVE: To explore the independent roles of pulse pressure and mean pressure as determinants of cardiovascular prognosis in older hypertensive patients. METHODS: This meta analysis, based on individual patient data, pooled the results of the European Working Party on High Blood Pressure in the Elderly trial (n = 840), the Systolic Hypertension in Europe Trial (n = 4695), and the Systolic Hypertension in China Trial (n = 2394). The relative hazard rates associated with pulse pressure and mean pressure were calculated using Cox regression analysis, with stratification for the 3 trials and with adjustments for sex, age, previous cardiovascular complications, smoking, and treatment group. RESULTS: A 10-mm Hg wider pulse pressure increased the risk of major cardiovascular complications; after controlling for mean pressure and the other covariates, the increase in risk ranged from approximately 13% for all coronary end points (P = .02) to nearly 20% for cardiovascular mortality (P = .001). In a similar analysis, mean pressure predicted the incidence of cardiovascular complications but only after removal of pulse pressure as an explanatory variable from the model. Furthermore, the probability of a major cardiovascular end point increased with higher systolic blood pressure; at any given level of systolic blood pressure, it also increased with lower diastolic blood pressure, suggesting that the wider pulse pressure was driving the risk of major complications. CONCLUSIONS: In older hypertensive patients, pulse pressure not mean pressure is the major determinant of cardiovascular risk. The implications of these findings for the management of hypertensive patients should be further investigated in randomized controlled outcome trials in which the pulsatile component of blood pressure is differently affected by antihypertensive drug treatment. PMID- 10789602 TI - Comparison of sustained-release nifedipine and temperature biofeedback for treatment of primary Raynaud phenomenon. Results from a randomized clinical trial with 1-year follow-up. AB - BACKGROUND: The efficacy and safety of sustained-release nifedipine for the treatment of primary Raynaud phenomenon (RP) has not previously been demonstrated by a randomized, controlled trial. Temperature biofeedback has been studied in patients with primary RP but not in a large multicenter controlled trial or compared with nifedipine therapy. OBJECTIVE: To evaluate and compare the effectiveness of sustained-release nifedipine and temperature biofeedback for the treatment of primary RP. PARTICIPANTS AND METHODS: This is a randomized, controlled clinical trial, double-masked for drug and placebo but not masked for temperature and control biofeedback. It included 313 persons with primary RP as defined by medical history, physical examination findings, normal nailfold capillaries, and a history of 2 or more attacks per day during the previous cold season. Participants were randomized to 1 of 4 treatment groups: (1) sustained release nifedipine, (2) pill placebo, (3) temperature biofeedback, or (4) control (electromyographic) biofeedback. The primary outcome measure was self-reported, color chart-verified RP attacks during 1 winter month approximately 1 year after initiation of treatment. Secondary outcome measures included verified attacks at 2 months, all attacks at 2 months and 1 year, and quality of life. RESULTS: Nifedipine-treated participants showed a 66% reduction in verified attacks compared with placebo recipients (P<.001); temperature biofeedback training did not reduce attacks significantly compared with control biofeedback (P = .37). Comparison of nifedipine and temperature biofeedback treatments favored nifedipine use (P = .08); similar results were obtained for the secondary end points. Adverse effects resulted in discontinuation of nifedipine treatment in 15% of participants. CONCLUSIONS: Temperature biofeedback is not better than its control treatment and is inferior to sustained-release nifedipine for treating primary RP, whereas sustained-release nifedipine is a safe and effective treatment for this disease. PMID- 10789601 TI - The risk of cardiovascular disease mortality associated with microalbuminuria and gross proteinuria in persons with older-onset diabetes mellitus. AB - BACKGROUND: Despite the numerous studies on the relation of albuminuria with increased risk of all-cause mortality in type 2 diabetes mellitus, it remains uncertain whether microalbuminuria and/or gross proteinuria are independent risk factors for cardiovascular mortality. Moreover, the association of albuminuria with cardiovascular mortality in people with type 2 diabetes mellitus has not been well described in US populations. OBJECTIVE: To estimate the relative risks (RRs) for the associations of microalbuminuria and gross proteinuria with cardiovascular disease mortality among persons with older-onset diabetes mellitus. METHODS: We conducted a prospective cohort study of 840 people with older-onset diabetes mellitus who provided urine samples in the 1984-1986 examination of a population-based study of diabetic persons. The presence of microalbuminuria was determined by an agglutination inhibition assay and gross proteinuria by a reagent strip. The main outcome was time to mortality from cardiovascular disease, as determined from death certificates. RESULTS: Of the 840 older-onset diabetic persons, 54.8% had normoalbuminuria, while 24.8% had microalbuminuria and 20.5% had gross proteinuria. During the 12-year follow-up (6127 person-years), we identified 364 deaths from cardiovascular disease. Compared with persons with normoalbuminuria, those with microalbuminuria and gross proteinuria had significantly higher risks of cardiovascular mortality. The RR as controlled for age, sex, glycemic control, insulin use, alcohol intake, physical activity, cardiovascular disease history, antihypertensive use, and retinopathy severity, was 1.84 (95% confidence interval [CI], 1.42-2.40) for those with microalbuminuria and 2.61 (95% CI, 1.99-3.43) for those with gross proteinuria. Further adjustment for other factors did not change the relations we found. When the end point used was mortality from coronary heart disease, stroke, or all causes, the increased risks were significant for both microalbuminuria (adjusted RRs [95% CIs], 1.96 [1.42-2.72], 2.20 [1.29-3.75], and 1.68 [1.35 2.09], respectively) and gross proteinuria (adjusted RRs [95% CIs], 2.73 [1.95 3.81], 2.33 [1.28-4.24], and 2.47 [1.97-3.10], respectively). CONCLUSIONS: Results from our population-based study strongly suggest that both microalbuminuria and gross proteinuria were significantly associated with subsequent mortality from all causes and from cardiovascular, cerebrovascular, and coronary heart diseases. These associations were independent of known cardiovascular risk factors and diabetes-related variables. PMID- 10789603 TI - "Benign" tumors and "early detection" in mammography-screened patients of a natural cohort with breast cancer. AB - BACKGROUND: Although the cure of breast cancer by "early detection" and prompt treatment rests on the belief that all breast cancers grow at the same rate, many cancers have been shown to grow rapidly and others slowly. In particular, mammography screening may often detect the slow-growing, nonaggressive tumors that might not be found until much later, if at all. METHODS: We reviewed the medical records of a natural cohort of 233 patients. The cohort comprised all women who received their first antineoplastic treatment for breast cancer at Yale New Haven Hospital during the period from January 1 through December 31, 1988, and had a median follow-up thereafter of 82.4 months. RESULTS: The mammography screen-detected group (MSDG) contained 97 (42%) of the 233 breast cancers. The rates of subsequent freedom from cancer deaths or recurrences were 95% (92 patients) in the MSDG and 79% (107 patients) in all other patients (log-rank 2P<.001). This superiority occurred partly because 90 (93%) of the MSDG were in the good prognosis TNM stages 0, I, and IIA, compared with 92 (68%) of the non MSDG (chi2 2P = .001). Of the 31 patients with stage 0 (carcinoma in situ), all of whom had disease-free survival, 24 (77%) were found by mammography screening. Even within similar TNM stages, however, the MSDG had distinctly better disease free survival results than the non-MSDG. For patients in TNM stages I and IIA, the "failure events" had respective rates of 2% and 13% (log-rank 2P = .02). CONCLUSIONS: The results suggest that many of the breast cancers found by mammography screening have excellent prognosis not just because of early detection, but also because many of the cancers are relatively benign, requiring minimal therapy. PMID- 10789604 TI - Yield of repeated screening for abdominal aortic aneurysm after a 4-year interval. Aneurysm Detection and Management Veterans Affairs Cooperative Study Investigators. AB - BACKGROUND: Little is known about the rate at which new abdominal aortic aneurysms (AAAs) develop or whether screening older men for AAA, if undertaken, should be limited to once in a lifetime or repeated at intervals. METHODS: A large population of veterans, aged 50 through 79 years, completed a questionnaire and underwent ultrasound screening for AAA. Of these, 5151 without AAA on the initial ultrasound (defined as infrarenal aortic diameter of 3.0 cm or larger) were selected randomly to be invited for a second ultrasound screening after an interval of 4 years. Local records and national databases were searched to identify deaths and AAA diagnoses made during the study interval in subjects who did not attend the rescreening. RESULTS: Of the 5151 subjects selected for a second screening, 598 (11.6%) had died (none due to AAA), and 20 (0.4%) had an interim diagnosis of AAA. A second screening was performed on 2622 (50.9%), of whom 58 (2.2%; 95% confidence interval, 1.6%-2.8%) had new AAA. Three new AAAs were 4.0 to 4.9 cm, 10 were 3.5 to 3.9 cm, and 45 were 3.0 to 3.4 cm. Independent predictors of new AAA at the second screening included current smoker (odds ratio, 3.09; 95% confidence, 1.74-5.50), coronary artery disease (odds ratio, 1.81; 95% confidence interval, 1.07-3.07), and, in a separate model using a composite variable, any atherosclerosis (odds ratio, 1.97; 95% confidence interval, 1.16-3.35). Adding the interim and rescreening diagnosis rates suggests a 4-year incidence rate of 2.6%. Rescreening only in subjects with infrarenal aortic diameter of 2.5 cm or greater on the initial ultrasound would have missed more than two thirds of the new AAAs. CONCLUSIONS: A second screening is of little practical value after 4 years, mainly because the AAAs detected are small. However, the incidence that we observed suggests that a second screening after longer intervals (ie, more than 8 years) may provide yields similar to those seen in initial screening and therefore warrants further study. PMID- 10789605 TI - Predictors of virological success and ensuing failure in HIV-positive patients starting highly active antiretroviral therapy in Europe: results from the EuroSIDA study. AB - BACKGROUND: Predictors of virological response to highly active antiretroviral therapy (HAART) have never been systematically evaluated in a large continental multicenter cohort of unselected human immunodeficiency virus (HIV)-infected people. OBJECTIVE: To determine the factors related to achieving and maintaining undetectable plasma HIV-1 RNA levels among HIV-1-infected patients first starting protease inhibitor- or nonnucleoside retrotranscriptase inhibitor-containing HAART in Europe. DESIGN: Prospective multicenter cohort study. SETTING: Fifty-two clinical centers in 17 European countries included in the EuroSIDA Study Group, from August 1996 to April 1999. PATIENTS: A total of 1469 HIV-positive patients first starting HAART recruited from an unselected cohort of more than 7300 HIV positive patients. MAIN OUTCOME MEASURE: Detection of factors related to virological success after first starting HAART (baseline) and ensuing failure by standard survival techniques, including Kaplan-Meier techniques and Cox proportional hazards models. All analyses were intention to treat. RESULTS: Most patients (80%) achieved plasma HIV-1 RNA levels of less than 500 copies/mL during follow-up (60.4% at 6 months from the onset of HAART). Patients with higher baseline HIV-1 RNA levels (relative hazard [RH], 0.76 per log higher; 95% confidence interval [CI], 0.69-0.84; P<.001) and those taking saquinavir mesylate hard gel as a single protease inhibitor (RH, 0.62; 95% CI, 0.47-0.82; P<.001) were less likely to reach undetectable HIV-1 RNA levels. Conversely, higher CD4+ lymphocyte counts (RH per 50% higher, 1.09; 95% CI, 1.02-1.16; P = .008) and the initiation of 3 or more new antiretroviral drugs (RH, 1.29; 95% CI, 1.03-1.61; P = .02) were independent predictors of higher success. Once success was achieved, HIV-1 RNA levels rebounded in more than one third of all patients during follow up (24% at 6 months). Antiretroviral-naive patients (RH, 0.50; 95% CI, 0.29-0.87; P = .01), older patients (RH, 0.86 per year older; 95% CI, 0.75-0.99; P = .04), and those starting a protease inhibitor other than saquinavir hard gel (RH, 0.66; 95% CI, 0.44-0.98; P = .04) were at decreased hazard for virological failure. Higher baseline HIV-1 RNA level (RH, 1.18 per log higher; 95% CI, 0.99-1.40; P = .06) and a longer time to achieve virological success (RH per 12 months, 1.53; 95% CI, 0.99-2.38; P = .06) were marginally significant predictors of a decreased hazard of ensuing virological failure. CONCLUSIONS: HAART is associated with a favorable virological response if started when the baseline HIV-1 RNA level is low, if at least 2 new nucleoside retrotranscriptase inhibitors are added, and if standard doses of saquinavir hard gel capsule are avoided as a single protease inhibitor. Older patients are more likely to achieve virological success. Thereafter, the higher durability of virological response is predicted by an antiretroviral-naive status and by the use of specific regimens. Lower baseline HIV-1 RNA levels and rapid maximal viral suppression seem to be other important factors in the durability of virological response. PMID- 10789606 TI - Effect of antiretroviral therapy on viral load, CD4 cell count, and progression to acquired immunodeficiency syndrome in a community human immunodeficiency virus infected cohort. Swiss HIV Cohort Study. AB - OBJECTIVE: To examine the effect of different antiretroviral treatment regimens on viral load, CD4 lymphocyte counts, and rates of progression to clinical acquired immunodeficiency syndrome events among treatment-naive human immunodeficiency virus (HIV)-infected patients enrolled in a large community cohort study. METHODS: Based in 7 outpatient clinics, the Swiss HIV Cohort Study is a cohort with national coverage. Virological, immunologic, and clinical results of 755 treatment-naive patients (median age, 36 years; 28.2% female) who initiated antiretroviral therapy between July 1, 1995, and June 30, 1997, were analyzed. Patients started undergoing monotherapy with 1 reverse transcriptase inhibitor (RTI), combination therapy with at least 2 RTIs, or highly active antiretroviral therapy (HAART) with RTIs and protease inhibitors. RESULTS: Antiretroviral treatment led to a mean reduction of viremia of 1.8 log10 copies per milliliter with HAART, 1.2 log10 copies per milliliter with RTI combination therapy, and 0.4 log10 copies per milliliter with monotherapy. Virological failure, defined as less than 1 log10 reduction per milliliter in viremia, was present in 45 (20%) patients undergoing HAART, 180 (38%) undergoing RTI combination therapy, and 47 (82%) undergoing monotherapy. The proportion of patients reaching undetectable viremia was 12% (n = 7) for monotherapy, 41% (n = 197) for RTI combination therapy, and 63% (n = 137) for HAART. Similar gains of CD4 cells were achieved with RTI combination therapy and HAART. Kaplan-Meier estimates of progression rates to a new acquired immunodeficiency syndrome event at 18 months were 13.6% (monotherapy), 4.7% (RTI combination therapy), and 3.9% (HAART). CONCLUSIONS: The rate of virological failure of antiretroviral treatments was high in this population of treatment-naive patients, even among patients receiving combination regimens. Clinical progression rates were, however, low in patients treated with RTI combination therapy and HAART. PMID- 10789607 TI - Patient comprehension and reaction to participating in a double-blind randomized clinical trial (ISIS-4) in acute myocardial infarction. AB - BACKGROUND: Although randomized clinical trials are currently the standard for the evaluation of new therapeutic strategies, little attention has been paid to the viewpoint of the patients recruited to these trials. OBJECTIVES: To examine the perspective of the Israeli patient cohort who participated in the Fourth International Study of Infarct Survival, a randomized trial in acute myocardial infarction. METHODS: A patient questionnaire was mailed to 360 Israeli patients who participated in the Fourth International Study of Infarct Survival and was returned by 150 of them. Main outcome measures included patient perception of consent procedures, comprehension of the study, subjective reaction to participating in the trial, and interest in present and future trials. RESULTS: Forty (31%) of 129 patients perceived that they had full comprehension of the trial, while 64 (50%) claimed partial understanding and 25 (19%), no understanding at all. Comprehension was related to a recollected explanation of 5 minutes or more (P<.001) and to an opportunity for discussion at the time of consent (P<.001). Most patients recollected the oral explanation; fewer, the written material. Patient consent was given by 64 (43%) of 150 patients in the hope of better treatment. In 36 cases (25%), the patients felt they received better treatment because of participation in the trial. CONCLUSIONS: Despite proper attention to accepted ethical and legal standards, perceived patient comprehension in this trial in acute myocardial infarction was incomplete or lacking in a considerable number of subjects. Much progress must be made toward the goal of true informed consent in clinical trials. PMID- 10789608 TI - Bundle-branch block as a risk factor in noncardiac surgery. AB - BACKGROUND: Despite extensive data examining perioperative risk in patients with coronary artery disease, little attention has been devoted to the implications of conduction system abnormalities. OBJECTIVE: To define the clinical significance of bundle-branch block (BBB) as a perioperative risk factor. METHODS: Retrospective, cohort-controlled study of all noncardiac, nonophthalmologic, adult patients with BBB seen in our preoperative evaluation center. Medical charts were reviewed for data regarding cardiovascular disease, surgical procedure, type of anesthesia, intravascular monitoring, and perioperative complications. RESULTS: Bundle-branch block was present in 455 patients. Right BBB (RBBB) was more common than left BBB (LBBB) (73.8% vs 26.2%). Three patients with LBBB and 1 patient with RBBB died; 1 patient had a supraventricular tachyarrhythmia. Three of the 4 deaths were sepsis related. There were 2 (0.4%) deaths in the control group. There was no difference in mortality between BBB and control groups (P = .32). Subgroup analysis suggested an increased risk for death in patients with LBBB vs controls (P = .06; odds ratio, 6.0; 95% confidence interval, 1.2-100.0) and vs RBBB (P = .06; odds ratio, 8.7; 95% confidence interval, 1.2-100.0). CONCLUSIONS: The presence of BBB is not associated with a high incidence of postoperative cardiac complications. Perioperative mortality is not increased in patients with RBBB and not directly attributable to cardiac complications in patients with LBBB. These data suggest that the presence of BBB does not significantly increase the likelihood of cardiac complications following surgery, but that patients with LBBB may not tolerate the stress of perioperative noncardiac complications. PMID- 10789609 TI - Serum lipid effects of a high-monounsaturated fat diet based on macadamia nuts. AB - BACKGROUND: Recent studies have identified potential beneficial effects of eating nuts, most of which have substantial amounts of monounsaturated fats. Macadamia nuts are 75% fat by weight, 80% of which is monounsaturated. OBJECTIVE: To examine variations in serum lipid levels in response to a high-monounsaturated fat diet based on macadamia nuts. METHODS: A randomized crossover trial of three 30-day diets was conducted in 30 volunteers aged 18 to 53 years from a free living population. Each was fed a "typical American" diet high in saturated fat (37% energy from fat); an American Heart Association Step 1 diet (30% energy from fat); and a macadamia nut-based monounsaturated fat diet (37% energy from fat) in random order. Serum total cholesterol, high-density lipoprotein cholesterol, and triglyceride levels were measured. RESULTS: Mean total cholesterol level after the typical American diet was 5.20 mmol/L (201 mg/dL). After the Step 1 diet and the macadamia nut diet, total cholesterol level was 4.99 mmol/L (193 mg/dL) and 4.95 mmol/L (191 mg/dL), respectively. Low-density lipoprotein cholesterol level was 3.37 mmol/L (130 mg/dL) (typical diet), 3.21 mmol/L (124 mg/dL) (Step 1 diet), and 3.22 mmol/L (125 mg/dL) (macadamia nut diet). High-density lipoprotein cholesterol level was 1.43 mmol/L (55 mg/dL) (typical), 1.34 mmol/L (52 mg/dL) (Step 1), and 1.37 mmol/L (53 mg/dL) (macadamia nut). Lipid values after the Step 1 and macadamia nut diets were significantly different from those after the typical diet (P<.05). CONCLUSIONS: The macadamia nut-based diet high in monounsaturated fat and the moderately low-fat diet both had potentially beneficial effects on cholesterol and low-density lipoprotein cholesterol levels when compared with a typical American diet. PMID- 10789610 TI - Hyperinsulinemia and the risk of cardiovascular death and acute coronary and cerebrovascular events in men: the Kuopio Ischaemic Heart Disease Risk Factor Study. AB - BACKGROUND: The role of hyperinsulinemia as a cardiovascular risk factor is controversial. We studied whether hyperinsulinemia is independently associated with increased cardiovascular morbidity and mortality. METHODS: Fasting serum insulin level and other cardiovascular risk factors were determined in 1521 men in eastern Finland aged 42 to 60 years with neither cardiovascular disease nor diabetes at baseline. Forty-five cardiovascular deaths, 110 acute coronary events, 48 strokes, and 163 any cardiovascular events occurred during an average follow-up of 9.5 years. A total of 163 cardiovascular events (45 cardiovascular deaths, 110 acute coronary events, and 48 strokes) occurred during an average follow-up of 9.5 years. RESULTS: In Cox regression analysis adjusting for age and examination years, fasting serum insulin level as a continuous variable was directly associated with the risk of cardiovascular death (P = .006), acute coronary events (P = .04), and stroke (P = .02). Men with insulin levels of 52 to 66 pmol/L, 67 to 89 pmol/L, and 90 pmol/L or more (3 highest quartiles) had 1.4 fold (95% confidence interval, 0.5-3.7), 1.4-fold (95% confidence interval, 0.5 3.7), and 2.5-fold (95% confidence interval, 1.0-5.9; P = .05) cardiovascular mortality, respectively, compared with men with insulin levels of less than 52 pmol/L (lowest quartile) (P = .04 for linear trend). Adjustment for serum lipid levels, blood pressure, and obesity reduced the excess cardiovascular mortality in the highest insulin quartile by 7%, 33%, and 67%, respectively. There were no statistically significant differences in the incidence of acute coronary events and stroke between the insulin quartiles. CONCLUSIONS: Hyperinsulinemia had a modest association with increased cardiovascular mortality in middle-aged men. This relationship was largely explained by obesity, hypertension, and dyslipidemia. Hyperinsulinemia had even weaker associations with the risk of acute coronary event and stroke. PMID- 10789611 TI - Multiple chemical sensitivity syndrome: symptom prevalence and risk factors in a military population. AB - OBJECTIVE: To assess the prevalence of and risk factors for self-reported symptoms suggestive of multiple chemical sensitivities/idiopathic environmental intolerance (MCS/IEI) in Persian Gulf War (PGW) veterans from Iowa and a comparison group of PGW-era military personnel. METHODS: A population-based sample of Iowa military personnel was surveyed using a cross-sectional telephone interview. Study participants were randomly drawn from 1 of 4 domains: PGW active duty, PGW National Guard/Reserve, non-PGW active duty, and non-PGW National Guard/Reserve. A complex sample survey design was used selecting participants from the following substrata: age, sex, race, rank, and military branch. The criteria for MCS/IEI were developed using expert consensus and the medical literature. RESULTS: A total of 3695 study participants (76% of those eligible) completed the telephone survey. The prevalence of symptoms suggestive of MCS/IEI in all participants was 3.4%. Veterans of the PGW reported a significantly higher prevalence of symptoms suggestive of MCS/IEI than did non-PGW military personnel (5.4% vs 2.6%); greater sensitivity to organic chemicals, vehicle exhaust, cosmetics, and smog; and more lifestyle changes. The following risk factors for MCS/IEI were identified with univariate analysis: deployment to the Persian Gulf, age (>25 years), female sex, receiving a physician diagnosis of MCS, previous professional psychiatric treatment, previous psychotropic medication use, current psychiatric illness, and a low level of preparedness. Multiple logistic regression analysis identified several independent risk factors for MCS/IEI, including deployment to the Persian Gulf, age, sex, rank, branch of service, previous professional psychiatric treatment, and current mental illness. CONCLUSIONS: Self-reported symptoms suggestive of MCS/IEI are relatively frequent in a military population and are more common among PGW veterans than comparable controls. Reported chemical sensitivities and accompanying behavioral changes were also frequent. After adjusting for age, sex, and training preparedness, previous professional psychiatric treatment and previous psychotropic medication use (before deployment) showed a robust association with symptoms suggestive of MCS. PMID- 10789612 TI - Extended-release niacin vs gemfibrozil for the treatment of low levels of high density lipoprotein cholesterol. Niaspan-Gemfibrozil Study Group. AB - OBJECTIVE: To provide a direct comparison of agents that raise plasma levels of high-density lipoprotein cholesterol (HDL-C) to help devise strategies for coronary risk reduction. METHODS: In a multicenter, randomized, double-blind trial, we compared the effects of extended-release niacin (Niaspan), at doses increased sequentially from 1000 to 2000 mg at bedtime, with those of gemfibrozil, 600 mg given twice daily, in raising low levels of HDL-C. Enrollment criteria included an HDL-C level of 1.03 mmol/L or less (< or =40 mg/dL), a low density lipoprotein cholesterol level of 4.14 mmol/L or less (< or =160 mg/dL) or less than 3.36 mmol/L (<130 mg/dL) with atherosclerotic disease, and a triglyceride level of 4.52 mmol/L or less (< or =400 mg/dL). RESULTS: Among 173 patients, 72 (82%) of the 88 assigned to Niaspan treatment and 68 (80%) of the 85 assigned to gemfibrozil treatment completed the study. Niaspan, at 1500 and 2000 mg, vs gemfibrozil raised the HDL-C level more (21% and 26%, respectively, vs 13%), raised the apolipoprotein A-I level more (9% and 11% vs 4%), reduced the total cholesterol-HDL-C ratio more (-17% and -22% vs -12%), reduced the lipoprotein(a) level (-7% and -20% vs no change), and had no adverse effect on the low-density lipoprotein cholesterol level (2% and 0% change vs a 9% increase). Significance levels for comparisons between medications ranged from P<.001 to P<.02. Gemfibrozil reduced the triglyceride level more than Niaspan (P<.001 to P = .06, -40% for gemfibrozil vs -16% to -29% for Niaspan, 1000 to 2000 mg). Effects on plasma fibrinogen levels were significantly favorable for Niaspan compared with gemfibrozil (P<.02), as gemfibrozil increased the fibrinogen level (from 5% to 9%) and Niaspan tended to decrease the fibrinogen level (from -1% to -6%). CONCLUSIONS: In patients with a low baseline HDL-C level, Niaspan at its higher doses provided up to 2-fold greater HDL-C increases, decreases in lipoprotein(a), improvements in lipoprotein cholesterol ratios, and lower fibrinogen levels compared with gemfibrozil. Gemfibrozil gave a greater triglyceride reduction but also increased the low-density lipoprotein cholesterol level, which did not occur with Niaspan. PMID- 10789613 TI - Evaluation of the Duke criteria in 93 episodes of prosthetic valve endocarditis: could sensitivity be improved? AB - BACKGROUND: Since publication of the Duke criteria for diagnosing endocarditis, several articles have confirmed their sensitivity when native and prosthetic valves are considered together. OBJECTIVES: To compare the differences between the older von Reyn criteria and the Duke criteria in prosthetic valve endocarditis only, and to determine if the latter's sensitivity could be improved by adding 2 minor criteria: new-onset heart failure and presence of conduction disturbances. METHODS: We retrospectively evaluated 93 episodes of prosthetic valve endocarditis from January 1986 to January 1998 in a teaching hospital, and then analyzed the 76 surgically confirmed episodes to compare the differences between the von Reyn and Duke diagnostic criteria. RESULTS: The von Reyn criteria rejected the diagnosis in 16 of the confirmed episodes, compared with 1 diagnosis missed by the Duke criteria and 1 missed using our suggested modifications. Definite diagnosis (Duke) was established in 60 episodes, compared with a diagnosis of probable (von Reyn) in 36 episodes (P<.001). Our modifications improved the sensitivity of the Duke criteria, diagnosing 70 episodes as definite (P = .02). CONCLUSIONS: As was the case with native valve endocarditis, the Duke criteria proved to be more sensitive than the von Reyn criteria in prosthetic valve endocarditis. The addition of 2 minor criteria (new-onset heart failure and presence of conduction disturbances) could improve the diagnostic sensitivity of the Duke criteria. PMID- 10789615 TI - Severe myopathy associated with vitamin D deficiency in western New York. AB - Five cases of severe myopathy associated with vitamin D deficiency are described. Each patient was confined to a wheelchair because of weakness and immobility. Two were elderly, 1 was a 37-year-old African American with type 1 diabetes mellitus, 1 was being treated for carcinoid syndrome, and 1 was severely malnourished due to poor oral intake. In each, weakness had previously been attributed to other causes, including old age, concomitant diabetic neuropathy, or general debility. Correct diagnosis was made initially by a high index of suspicion, following the demonstration of clinical proximal myopathy; confirmation was made by the demonstration of low 25-hydroxyvitamin D and elevated parathyroid hormone concentrations. Treatment with vitamin D caused a resolution of body aches and pains and a restoration of normal muscle strength in 4 to 6 weeks. Four patients became fully mobile and had normal 25-hydroxyvitamin D concentrations, and the fifth also became mobile. In the 4 fully recovered cases, parathyroid hormone levels on follow-up were lower but still elevated. This finding suggests a degree of autonomy of parathyroid secretion known to occur in cases of long-standing vitamin D deficiency. Myopathy, due to chronic vitamin D deficiency, probably contributes to immobility and ill health in a significant number of patients in the northern United States. An awareness of this condition may significantly improve mobility and quality of life in patient populations vulnerable to vitamin D deficiency. PMID- 10789614 TI - Systematic review of the epidemiological evidence on Helicobacter pylori infection and nonulcer or uninvestigated dyspepsia. AB - BACKGROUND: Previous studies have yielded conflicting results and substantial uncertainty about any independent association of Helicobacter pylori infection with dyspepsia, and about any benefits of antibiotic treatments for nonulcer or uninvestigated dyspepsia. OBJECTIVES: To perform a systematic review of the literature to determine whether chronic infection with H pylori is relevant to nonulcer or uninvestigated dyspepsia. METHODS: Observational studies of associations between H pylori and dyspepsia published before April 1999 and randomized trials of the effects of H pylori eradication on dyspepsia published before January 2000 were identified by computer-assisted literature searches of relevant journals, reference lists, and discussions with authors. Relevant data were abstracted from the published reports by 2 investigators according to a fixed protocol. RESULTS: Thirty relevant observational studies were identified involving approximately 3392 patients with nonulcer dyspepsia, and 11 separate observational studies were identified, involving 6426 patients with uninvestigated dyspepsia. Reports of strong associations in small observational studies without appropriate adjustment for potential confounding factors were not generally confirmed by larger and better-designed studies. No studies have been reported, however, that can reliably confirm or exclude the existence of any weak associations. Twenty-two randomized trials of treatments against H pylori were found involving a total of 2340 patients with nonulcer dyspepsia, almost all with positive H pylori test results. Only a few of these trials involved effective antibacterial regimens with prolonged follow-up, and even these studies were too small to assess the possibility of moderate benefits. CONCLUSION: The available evidence indicates that there is no strong association between H pylori and dyspepsia, but there is insufficient evidence to confirm or refute the existence of a modest association. PMID- 10789616 TI - Prostate cancer screening in high-risk patients. PMID- 10789618 TI - The power of the mind. PMID- 10789617 TI - Mind-body misunderstandings. PMID- 10789619 TI - Integrative medicine: who needs it and why? PMID- 10789620 TI - Another round of jeopardy. PMID- 10789621 TI - An historical take on the physician's charter. PMID- 10789622 TI - Studying the statins. PMID- 10789623 TI - Degrees of difficulty in ascertaining credentials. PMID- 10789624 TI - Medicare. PMID- 10789625 TI - Medicare. PMID- 10789626 TI - An outbreak of hepatitis B associated with reusable subdermal electroencephalogram electrodes. Hepatitis B Outbreak Investigation Team. AB - BACKGROUND: In early 1996 an outbreak of hepatitis B was detected among patients who attended an electroencephalogram (EEG) clinic in Toronto operated by a neurologist. In this article we report the results of an investigation conducted to determine the extent and source of the outbreak. METHODS: Notifications were sent to 18 567 patients who had attended any of 6 EEG clinics operated by the neurologist between 1990 and 1996 asking them to see their physician to be tested for hepatitis B virus (HBV) infection; 2957 envelopes were returned. Of the remaining 15 610 patients, results of laboratory tests were available for 10 244 (65.6%). A detailed follow-up of patients with newly acquired hepatitis B and those with chronic infection (carriers) was conducted. Viral DNA sequencing was used to compare strains of available HBV isolates. RESULTS: A total of 75 patients were identified in whom hepatitis B developed between 1991 and 1996; all of them had had at least one EEG performed in which reusable subdermal electrodes had been used. No cases were detected among patients who participated only in sleep studies, for which disk electrodes had been used. The peak rate of HBV infection (18.2 cases per 1000 person-EEGs) occurred in 1995. One technician performed all of the EEGs at the clinics and was found to be positive for hepatitis B e antigen. DNA sequencing confirmed that the virus isolated from the technician was identical to the virus isolated in 4 cases of hepatitis B tested. Infection control procedures were found to be inadequate. INTERPRETATION: The hepatitis B outbreak was a result of a common source of infection, the technologist, and inadequate infection control practices. Reusable subdermal EEG electrodes were the likely vehicles of transmission. Health care workers should follow recommended infection control practices and be vaccinated against hepatitis B. PMID- 10789627 TI - Surveillance for outbreaks of respiratory tract infections in nursing homes. AB - BACKGROUND: Outbreaks of respiratory tract infections are common in long-term care facilities for older people. The objective of our study was to determine both the frequency of such outbreaks and their clinical and epidemiological features. METHODS: Prospective surveillance for outbreaks of respiratory tract infections and a retrospective audit of surveillance records were conducted in 5 nursing homes in metropolitan Toronto over 3 years. The clinical manifestations of infected residents were identified and microbiological investigations for causal agents were conducted. RESULTS: Sixteen outbreaks, involving 480 of 1313 residents, were identified prospectively during 1 144 208 resident-days of surveillance, for an overall rate of 0.42 infections per 1000 resident-days. Another 30 outbreaks, involving 388 residents, were identified retrospectively. Outbreaks occurred year-round, with no seasonal pattern. Pathogens included influenza virus, parainfluenza virus, respiratory syncytial virus, Legionella sainthelensi and Chlamydia pneumoniae. Multiple pathogens were detected in 38% (6/16) of the prospectively identified outbreaks. Of the 480 residents in the prospectively identified outbreaks 398 (83%) had a cough, 194 (40%) had fever and 215 (45%) had coryza. Clinical findings were nonspecific and could not be used to distinguish between causal agents. Pneumonia developed in 72 (15%) of the 480 residents, and 58 (12%) required transfer to hospital. The case-fatality rate was 8% (37/480). INTERPRETATION: Our findings emphasize the importance of adequate surveillance for outbreaks of respiratory tract infections in nursing homes and of early diagnosis so that appropriate interventions can be promptly instituted. PMID- 10789628 TI - Biochemical markers in acute ischemic stroke. PMID- 10789629 TI - Changing drinking-and-driving behaviour: the effects of Ontario's administrative driver's licence suspension law. PMID- 10789630 TI - Ethics through the looking glass. PMID- 10789631 TI - High performance computing and medical research. PMID- 10789632 TI - Reducing medication errors. PMID- 10789633 TI - The silent payer speaks: workers' compensation boards and Canadian physicians. PMID- 10789634 TI - Rheumatology: 2. What laboratory tests are needed? PMID- 10789635 TI - Ethics and human rights in South African medicine. PMID- 10789636 TI - Health care, federalism and the new Social Union. AB - The Social Union framework agreement and the Health Accord provide examples of the close relationship that exists between federalism and the delivery of health care. These recent agreements represent a move from a federal-unilateral style of federalism to a more collaborative model. This shift will potentially affect federal funding for health care, interpretation of the Canada Health Act and the development of new health care initiatives. The primary advantage of the new collaborative model is protection of jurisdictional autonomy. Its primary disadvantages are blurring of accountability and potential for exclusion of the public from decision-making. PMID- 10789637 TI - Doctors take NB government to court. PMID- 10789638 TI - Cataract surgery contracted out in Vancouver. PMID- 10789639 TI - Delay in cancer centre expansion may derail Ontario project. PMID- 10789640 TI - BC's reference-based pricing stirs controversy. PMID- 10789641 TI - An atlas of the brain, online. PMID- 10789642 TI - Availability of services in rural areas. PMID- 10789643 TI - Hypertrophic pyloric stenosis caused by erythromycin. PMID- 10789644 TI - He said, she said: demystifying the dreaded disciplinary hearing. PMID- 10789645 TI - UBC med school marks 50th birthday with call for increased enrolment. PMID- 10789646 TI - Can the Senate save medicare? PMID- 10789647 TI - A piece of my mind. The stories. PMID- 10789648 TI - Polymer scientists engineer better remedies. PMID- 10789649 TI - Final report on mechanical vs bioprosthetic heart valves. PMID- 10789650 TI - Japanese develop early test for lung cancer metastases. PMID- 10789651 TI - Health agencies update. PMID- 10789652 TI - From the Centers for Disease Control and Prevention. Update: pulmonary hemorrhage/hemosiderosis among infants--Cleveland, Ohio, 1993-1996. PMID- 10789654 TI - From the Centers for Disease Control and Prevention. Availability of Work-Related Lung Disease Surveillance Report, 1999 PMID- 10789653 TI - From the Centers for Disease Control and Prevention. Imported dengue--United States, 1997 and 1998. PMID- 10789655 TI - Sodium intake as a risk factor for cardiovascular disease. PMID- 10789656 TI - Sodium intake as a risk factor for cardiovascular disease. PMID- 10789657 TI - Smoking and adverse childhood experiences. PMID- 10789658 TI - Smoking and adverse childhood experiences. PMID- 10789660 TI - Rofecoxib and the risk of adverse upper gastrointestinal effects. PMID- 10789659 TI - Rofecoxib and the risk of adverse upper gastrointestinal effects. PMID- 10789661 TI - Cyclooxygenase 2 selective agents and upper gastrointestinal disease. PMID- 10789662 TI - Cyclooxygenase 2 selective agents and upper gastrointestinal disease. PMID- 10789663 TI - NIH research grants: funding and re-funding. PMID- 10789664 TI - Major cardiovascular events in hypertensive patients randomized to doxazosin vs chlorthalidone: the antihypertensive and lipid-lowering treatment to prevent heart attack trial (ALLHAT). ALLHAT Collaborative Research Group. AB - CONTEXT: Hypertension is associated with a significantly increased risk of morbidity and mortality. Only diuretics and beta-blockers have been shown to reduce this risk in long-term clinical trials. Whether newer antihypertensive agents reduce the incidence of cardiovascular disease (CVD) is unknown. OBJECTIVE: To compare the effect of doxazosin, an alpha-blocker, with chlorthalidone, a diuretic, on incidence of CVD in patients with hypertension as part of a study of 4 types of antihypertensive drugs: chlorthalidone, doxazosin, amlodipine, and lisinopril. DESIGN: Randomized, double-blind, active-controlled clinical trial, the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial, initiated in February 1994. In January 2000, after an interim analysis, an independent data review committee recommended discontinuing the doxazosin treatment arm based on comparisons with chlorthalidone. Therefore, outcomes data presented herein reflect follow-up through December 1999. SETTING: A total of 625 centers in the United States and Canada. PARTICIPANTS: A total of 24,335 patients (aged > or = 55 years) with hypertension and at least 1 other coronary heart disease (CHD) risk factor who received either doxazosin or chlorthalidone. INTERVENTIONS: Participants were randomly assigned to receive chlorthalidone, 12.5 to 25 mg/d (n=15,268), or doxazosin, 2 to 8 mg/d (n=9067), for a planned follow-up of 4 to 8 years. MAIN OUTCOME MEASURES: The primary outcome measure was fatal CHD or nonfatal myocardial infarction (MI), analyzed by intent to treat; secondary outcome measures included all-cause mortality, stroke, and combined CVD (CHD death, nonfatal MI, stroke, angina, coronary revascularization, congestive heart failure [CHF], and peripheral arterial disease); compared by the chlorthalidone group vs the doxazosin group. RESULTS: Median follow-up was 3.3 years. A total of 365 patients in the doxazosin group and 608 in the chlorthalidone group had fatal CHD or nonfatal MI, with no difference in risk between the groups (relative risk [RR], 1.03; 95% confidence interval [CI], 0.90-1.17; P=.71). Total mortality did not differ between the doxazosin and chlorthalidone arms (4-year rates, 9.62% and 9.08%, respectively; RR, 1.03; 95% CI, 0.90-1.15; P=.56.) The doxazosin arm, compared with the chlorthalidone arm, had a higher risk of stroke (RR, 1.19; 95% CI, 1.01-1.40; P=.04) and combined CVD (4-year rates, 25.45% vs 21.76%; RR, 1.25; 95% CI, 1.17 1.33; P<.001). Considered separately, CHF risk was doubled (4-year rates, 8.13% vs 4.45%; RR, 2.04; 95% CI, 1.79-2.32; P<.001); RRs for angina, coronary revascularization, and peripheral arterial disease were 1.16 (P<.001), 1.15 (P=.05), and 1.07 (P=.50), respectively. CONCLUSION: Our data indicate that compared with doxazosin, chlorthalidone yields essentially equal risk of CHD death/nonfatal MI but significantly reduces the risk of combined CVD events, particularly CHF, in high-risk hypertensive patients. PMID- 10789665 TI - The relationship between managed care insurance and use of lower-mortality hospitals for CABG surgery. AB - CONTEXT: Explicit information about the quality of coronary artery bypass graft (CABG) surgery has been available for nearly a decade in New York State; however, the extent to which managed care insurance plans direct enrollees to the lowest mortality CABG surgery hospitals remains unknown. OBJECTIVE: To compare the proportion of patients with managed care insurance and fee-for-service (FFS) insurance who undergo CABG surgery at lower-mortality hospitals. DESIGN: A retrospective cohort study of CABG surgery discharges from 1993 to 1996, using New York Department of Health databases and multivariate analysis to estimate the use of lower-mortality hospitals by patients with different types of health insurance. SETTING: Cardiac surgical centers in New York, of which 14 were classified as lower-mortality hospitals (mean rate, 2.1%) and 17 were classified as higher-mortality hospitals (mean rate, 3.2%). PATIENTS: A total of 58,902 adults older than 17 years who were hospitalized for CABG surgery. Patients were excluded if their CABG surgery was combined with any valve procedure or left ventricular aneurysm resection or if they were younger than 65 years and enrolled in Medicare FFS or Medicare managed care. MAIN OUTCOME MEASURE: Probability of a patient receiving CABG surgery at a lower-mortality hospital. RESULTS: Compared with patients with private FFS insurance (n = 18,905), patients with private managed care insurance (n=7169) and Medicare managed care insurance (n=880) were less likely to receive CABG surgery at a lower-mortality hospital (relative risk [RR] of surgery at a lower-mortality hospital compared with patients with private FFS insurance, 0.77; 95% confidence interval [CI], 0.74-0.81; P<.001; and RR, 0.61; 95% CI, 0.54-0.70; P<.001, respectively, after controlling for multiple potential confounding factors). Patients with Medicare FFS insurance used lower mortality hospitals at rates more similar to those with private FFS insurance (n = 31,948; RR, 0.95; 95% CI, 0.91-0.98; P=.004). CONCLUSIONS: Patients in New York State with private managed care and Medicare managed care insurance were significantly less likely to use lower-mortality hospitals for CABG surgery compared with patients with private FFS insurance. PMID- 10789666 TI - Enduring effects of nurse home visitation on maternal life course: a 3-year follow-up of a randomized trial. AB - CONTEXT: A home visitation program using nurses to improve maternal and child outcomes had favorable results in a randomized trial with a primarily white, semirural population. Many of the short-term findings have been replicated with urban blacks, but whether the program will continue to demonstrate effectiveness after its conclusion is uncertain. OBJECTIVE: To determine the effectiveness of a prenatal and infancy home visitation program on the maternal life course of women in an urban environment 3 years after the program ended. DESIGN AND SETTING: Three-year follow-up of a randomized controlled trial of women seen consecutively between June 1990 and August 1991 at an obstetrical clinic in Memphis, Tenn, who were enrolled in a visitation program for 2 years after the birth of their first child. PARTICIPANTS: A cohort of 743 women who were primarily black, were pregnant for less than 29 weeks, had no previous live births, and had at least 2 sociodemographic risk factors (unmarried, <12 years of education, or unemployed). INTERVENTION: An average of 7 (range, 0-18) home visits during pregnancy and 26 (range, 0-71) from birth to the child's second birthday. MAIN OUTCOME MEASURES: Rate of subsequent pregnancy, mean interval between first and second birth, and mean number of months of welfare use. RESULTS: Compared with the control group, women who received home visits by nurses had fewer subsequent pregnancies (1.15 vs 1.34; P=.03), fewer closely spaced subsequent pregnancies (0.22 vs 0.32; P=.03), longer intervals between the birth of the first and second child (30.25 vs 26.60 months; P=.004), and fewer months of using Aid to Families with Dependent Children (32.55 vs 36.19; P=.01) and food stamps (41.57 vs 45.04; P=.005). Compared with the effect of the program while the program was in operation, the effect after it ended was essentially equal for Aid to Families with Dependent Children, greater for food stamps, greater for rates of closely spaced subsequent pregnancies, and smaller for rates of subsequent pregnancy overall. CONCLUSIONS: We found enduring effects of a home visitation program on the lives of black women living in an urban setting. While these results were smaller in magnitude than those achieved in a previous trial with white women living in a semirural setting, the direction of the effects was consistent across the 2 studies. PMID- 10789667 TI - The effect of organized systems of trauma care on motor vehicle crash mortality. AB - CONTEXT: Despite calls for wider national implementation of an integrated approach to trauma care, the effectiveness of this approach at a regional or state level remains unproven. OBJECTIVE: To determine whether implementation of an organized system of trauma care reduces mortality due to motor vehicle crashes. DESIGN: Cross-sectional time-series analysis of crash mortality data collected for 1979 through 1995 from the Fatality Analysis Reporting System. SETTING: All 50 US states and the District of Columbia. SUBJECTS: All front-seat passenger vehicle occupants aged 15 to 74 years. MAIN OUTCOME MEASURES: Rates of death due to motor vehicle crashes compared before and after implementation of an organized trauma care system. Estimates are based on within-state comparisons adjusted for national trends in crash mortality. RESULTS: Ten years following initial trauma system implementation, mortality due to traffic crashes began to decline; about 15 years following trauma system implementation, mortality was reduced by 8% (95% confidence interval [CI], 3%-12%) after adjusting for secular trends in crash mortality, age, and the introduction of traffic safety laws. Implementation of primary enforcement of restraint laws and laws deterring drunk driving resulted in reductions in crash mortality of 13% (95% CI, 11%-16%) and 5% (95% CI, 3%-7%), respectively, while relaxation of state speed limits increased mortality by 7% (95% CI, 3%-10%). CONCLUSIONS: Our data indicate that implementation of an organized system of trauma care reduces crash mortality. The effect does not appear for 10 years, a finding consistent with the maturation and development of trauma triage protocols, interhospital transfer agreements, organization of trauma centers, and ongoing quality assurance. PMID- 10789668 TI - Ketoconazole for early treatment of acute lung injury and acute respiratory distress syndrome: a randomized controlled trial. The ARDS Network. AB - CONTEXT: Three clinical studies have suggested that ketoconazole, a synthetic imidazole with anti-inflammatory activity, may prevent the development of acute respiratory distress syndrome (ARDS) in critically ill patients. However, the use of ketoconazole as treatment for acute lung injury (ALI) and ARDS has not been previously studied. OBJECTIVE: To test the efficacy of ketoconazole in reducing mortality and morbidity in patients with ALI or ARDS. DESIGN: Randomized, double blind, placebo-controlled trial conducted from March 1996 to January 1997. SETTING: Twenty-four hospitals associated with 10 network centers in the United States, constituting the ARDS Network. PATIENTS: A total of 234 patients with ALI or ARDS. INTERVENTION: Patients were randomly assigned to receive ketoconazole, 400 mg/d (n = 117), or placebo (n = 117), initiated within 36 hours of fulfilling study entry criteria and given enterally for up to 21 days. MAIN OUTCOME MEASURES: Primary outcome measures were the proportion of patients alive with unassisted breathing at hospital discharge and the number of days of unassisted breathing (ventilator-free days) during 28 days of follow-up. Secondary outcome measures included the proportion of patients achieving unassisted breathing for 48 hours or more, the number of organ failure-free days, and changes in plasma interleukin 6 (IL-6) and urinary thromboxane A2 metabolites (thromboxane B2 [TXB2] and 11-dehydro-TXB2). RESULTS: In-hospital mortality (SE) was 34.1% (4.3%) for the placebo group and 35.2% (4.3%) for the ketoconazole group (P=.85). The median number of ventilator-free days within 28 days of randomization was 9 in the placebo group and 10 in the ketoconazole group (P=.89). There were no statistically significant differences in the number of organ failure-free days, pulmonary physiology, or adverse events between treatment groups. The median serum ketoconazole level was 1.25 microg/mL and serum levels greater than 0.5 microg/mL were detected in 96% of patients assayed. Plasma IL-6, urinary TXB2, and 11-dehydro-TXB2 levels were unaffected by ketoconazole. CONCLUSIONS: In these patients with ALI or ARDS, ketoconazole was safe and bioavailable but did not reduce mortality or duration of mechanical ventilation or improve lung function. These data do not support the use of ketoconazole for the early treatment of ALI or ARDS. PMID- 10789669 TI - The case against anergy testing as a routine adjunct to tuberculin skin testing. AB - Although anergy testing is commonly used to help interpret negative tuberculin skin test results, the validity of this approach has not been demonstrated. Specific issues include lack of a standardized protocol for antigen selection, number needed to reliably evaluate inability to respond, and uniform criteria for defining cutaneous reactivity, as well as regional variation in skin test reactivity. Tuberculin skin testing is used to screen for latent infection and to evaluate the need for isoniazid prophylaxis. The presence or absence of reactivity to control antigens does not affect this decision. The results of anergy testing also do not predict the risk for progression to active disease in either HIV-negative or HIV-positive patients. In HIV-negative patients with active tuberculosis, 10% to 20% have negative tuberculin test results, and 5% to 10% have a negative tuberculin result but have a positive reaction to another antigen. A negative tuberculin skin test result does not exclude either latent infection or active disease, even in the presence of a reaction to other antigens. Neither anergy testing nor tuberculin testing obviates the need for microbiologic evaluation when there is suspicion for active tuberculosis infection. Therefore, anergy testing is not useful in screening for asymptomatic tuberculous infection or for diagnosing active tuberculosis. PMID- 10789671 TI - Diuretics vs alpha-blockers for treatment of hypertension: lessons from ALLHAT. Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial. PMID- 10789670 TI - Meta-analysis of observational studies in epidemiology: a proposal for reporting. Meta-analysis Of Observational Studies in Epidemiology (MOOSE) group. AB - OBJECTIVE: Because of the pressure for timely, informed decisions in public health and clinical practice and the explosion of information in the scientific literature, research results must be synthesized. Meta-analyses are increasingly used to address this problem, and they often evaluate observational studies. A workshop was held in Atlanta, Ga, in April 1997, to examine the reporting of meta analyses of observational studies and to make recommendations to aid authors, reviewers, editors, and readers. PARTICIPANTS: Twenty-seven participants were selected by a steering committee, based on expertise in clinical practice, trials, statistics, epidemiology, social sciences, and biomedical editing. Deliberations of the workshop were open to other interested scientists. Funding for this activity was provided by the Centers for Disease Control and Prevention. EVIDENCE: We conducted a systematic review of the published literature on the conduct and reporting of meta-analyses in observational studies using MEDLINE, Educational Research Information Center (ERIC), PsycLIT, and the Current Index to Statistics. We also examined reference lists of the 32 studies retrieved and contacted experts in the field. Participants were assigned to small-group discussions on the subjects of bias, searching and abstracting, heterogeneity, study categorization, and statistical methods. CONSENSUS PROCESS: From the material presented at the workshop, the authors developed a checklist summarizing recommendations for reporting meta-analyses of observational studies. The checklist and supporting evidence were circulated to all conference attendees and additional experts. All suggestions for revisions were addressed. CONCLUSIONS: The proposed checklist contains specifications for reporting of meta-analyses of observational studies in epidemiology, including background, search strategy, methods, results, discussion, and conclusion. Use of the checklist should improve the usefulness of meta-analyses for authors, reviewers, editors, readers, and decision makers. An evaluation plan is suggested and research areas are explored. PMID- 10789672 TI - Clinical performance measurement--a hard sell. PMID- 10789673 TI - JAMA Patient Page: tuberculosis. PMID- 10789674 TI - Neutrophil NADPH oxidase does not assemble on macropinocytic vacuole membranes. AB - NADPH oxidase assembly is considered to occur mainly on the plasma membrane and phagolysosome membranes and to a lesser extent in intracellular granules. Stimulation of neutrophils with phorbol 12-myristate 13-acetate (PMA) in conjunction with chemiluminescent substrates has been widely used as a model to understand the sub-cellular locations of NADPH-oxidase activity. Interpretation of data from such studies is complicated by observations that PMA does not trigger phagocytosis but results in formation of large macropinocytic vacuoles. Here we show by laser-scanning confocal microscopy that PMA triggers uptake of lucigenin into macropinocytic vacuoles and that no chemiluminescence (CL) can be detected at this location when NADPH oxidase assembly on the plasma membrane still occurs. This shows that the macropinocytic vacuole membrane is distinct from the phagocytic vacuole membrane although both compartments are derived from the plasma membrane. PMID- 10789676 TI - Normal immune function in young and old DNA polymerase-beta deficient mice. AB - The effect of the DNA polymerase-beta (beta-pol) deficiency on mitogenic response and cytokine production was studied in spleen lymphocytes from 4-5- and 20-22 month-old beta-pol(-/+) mice and their age-matched wild-type littermates. The proliferative response of lymphocytes to Concanavalin A (Con A) and lipopolysaccharide (LPS) was measured by [3H]thymidine incorporation, and the induction of cytokine production (interleukin (IL)-2, IL-4, and interferon necrosis factor (IFN)-gamma) was assessed by enzyme-linked immunosorbent assay. There was no significant difference in Con A- or LPS-induced proliferation or cytokine production in young beta-pol(-/+) mice compared with young wild-type littermates or in old beta-pol(-/+) mice compared with old wild-type littermates. However, mitogen-induced proliferation and cytokine production changed significantly with age. The proliferative response to Con A and to LPS, and the IL-2 production was significantly lower, and IL-4 and IFN-gamma levels were significantly higher in lymphocytes from old beta-pol(-/+) mice and old wild-type mice than in lymphocytes from young beta-pol(-/+) mice and young wild-type littermates. In addition, flow cytometric analysis showed no significant differences between young beta-pol(-/+) mice and young wild-type littermates or between old beta-pol(-/+) mice and old wild-type littermates in the proportion of B- and T-cell populations, and T-cell subsets. However, the number of lymphocytes expressing CD4+ phenotype slightly decreased and the proportion of lymphocytes expressing CD44/Pgp-1 (memory) phenotype increased with age. Thus, we found no evidence for alteration in immune function in DNA polymerase-beta deficient mice, although they exhibit a decline in immunologic function with age. PMID- 10789675 TI - The SH2 domain containing inositol 5-phosphatase SHIP2 associates to the immunoreceptor tyrosine-based inhibition motif of Fc gammaRIIB in B cells under negative signaling. AB - Fc gammaRIIB are single-chain low-affinity receptors for IgG that bear an immunoreceptor tyrosine-based inhibition motif (ITIM) in their intracytoplasmic domain and that negatively regulate immunoreceptor tyrosine-based activation motif (ITAM)-dependent cell activation. In B cells, coaggregation of the B cell receptor (BCR) and Fc gammaRIIB leads to an inhibition of B cell activation. Inhibitory properties of Fc gammaRIIB have been related to the recruitment of SHIP, an SH2 domain-containing inositol 5-phosphatase (referred to as SHIP1), via ITIM phosphorylated Fc gammaRIIB. Here, we demonstrate that the second SH2 domain containing inositol 5-phosphatase SHIP2 could also bind to the Fc gammaRIIB ITIM. As a model, a Fc gammaRIIB deficient B cell line (IIA1.6), transfected with a cDNA encoding either w.t. Fc gammaRIIB1' or Fc gammaRIIB1' whose ITIM tyrosine was mutated has been used. SHIP2 tyrosine phosphorylation and association to the adaptator protein Shc were only found in transfectants expressing w.t. Fc gammaRIIB1'. SHIP2 was also found to bind to a phosphopeptide corresponding to the ITIM sequence of Fc gammaRIIB. There was no binding to the nonphosphorylated peptide. Finally, both SHIP2 and SHIP1 were coprecipitated with Fc gammaRIIB1' upon coaggregation with BCR in IIA1.6 transfectants. PMID- 10789677 TI - Apoptosis of murine thymocytes induced by extracellular ATP is dose- and cytosolic pH-dependent. AB - Thymocytes from young Balb/C mice responded to low extracellular ATP (ATPec) doses (< or = 0.3 mM) with a rapid intracellular acidification (mean pH: ca. 0.3 pH unit) that was inhibited by the Ca2+ channel blocker verapamil, or by suramin (50 microM) and TNP-ATP (40 microM), potent P2x (and P2y) purinoreceptor antagonists. ATPec also triggered a remarkable DNA fragmentation and cell shrinkage detectable only at these low doses. DNA fragmentation gradually disappears with increasing [ATPec] above 0.5 mM, with a concomitant dominance of cytosolic alkalinization of the cells. Suramin and TNP-ATP also blocked the ATPec triggered DNA fragmentation efficiently. oATP, inhibitor of P2z nonspecific ATP gated membrane pores, and 2 mM extracellular Mg2+ did not influence either the cytosolic acidification or the DNA fragmentation, but almost completely abolished the intracellular alkalinization characteristic of P2z receptor activation at high ATPec doses. Antagonist-sensitivity of the ATPec-induced membrane potential responses indicates that hyperpolarization is associated with intracellular acidification, while rapid depolarization is linked to alkalinization. These data together indicate that the Ca2+-dependent hyperpolarization and cytosolic acidification triggered by low ATPec doses are essential early signals in apoptosis of murine thymocytes and are likely mediated by P2x1 type ATP-gated ion channels. Subset specificity of the early purinergic signals suggests that the double positive thymocytes are most sensitive to ATPec showing both P2z and P2x receptor activation characteristics, the double negative thymocytes preferentially show P2z-type, while single positive (CD4- CD8+ or CD4+ CD8-) thymocytes respond mostly by weaker P2x-type changes, indicating that ATPec, similarly to adenosine may serve as a potential regulator of cell death and differentiation in the thymus. PMID- 10789678 TI - Cetirizine-induced downregulation of airway fibroblast proliferation and function: a rationale for a different approach to allergy treatment? AB - Recently, airway fibroblasts captured the attention of both allergists and basic scientists since they are no longer considered as mere bystanders, as far as allergic airway diseases are concerned. The aim of the present study was to assess the effects of different Cetirizine (Cet) concentrations (0.01, 0.05, 0.1 mg/ml) on human airway fibroblast proliferation and on CD54 expression. By means of flow cytometry analysis, we evaluated CD54 expression by airway fibroblasts in basal conditions or after gammaIFN stimulation in the presence of Cetirizine; we also evaluated the effect of the drug on cell proliferation by a [3H]thymidine incorporation assay. All of the tested doses of Cetirizine were able to significantly reduce CD54 upregulation induced by gammaIFN; concerning the fibroblast proliferation, we observed a dose-dependent inhibition of [3H]thymidine incorporation. These results show that Cetirizine exerts a biologic effect directly on human airway fibroblasts, suggesting a new rationale in the use of this compound. PMID- 10789679 TI - Gammadelta+ T cells and 65-kDa heat shock protein expression following Cryptosporidium parvum challenge in athymic C57BL/6J nude mice. PMID- 10789680 TI - Induction of apoptosis in bone marrow cells by gangliosides produced by a T cell lymphoma. AB - The present study was conducted to investigate the mode of cell death of bone marrow cells (BMC) treated in vivo or in vitro with gangliosides (DLG) derived from a T cell lymphoma, designated as Dalton's lymphoma (DL). BMC undergoing cell death showed morphological features characteristic to apoptosis such as cell shrinkage, plasma membrane blebbing and densely stained chromatin. DLG-induced apoptosis was further confirmed by analysis of DNA from DLG-treated BMC by flow cytometry and agarose gel electrophoresis which showed an increase in hypodiploid DNA and internucleosomal DNA cleavage, respectively. Immunoblotting of p53 and Bax demonstrated an increase in the levels of p53 and Bax proteins. BMC treated with DLG also showed translocation of NF-kappaB protein into the nucleus. Antibodies to ganglioside G(D3), a constituent of DLG, neutralized the effect of DLG. Further, supplementation of DLG-containing BMC cultures with granulocyte macrophage-colony stimulating factor (GM-CSF) or macrophage-colony stimulating factor (M-CSF) prevented the apoptosis-inducing action of DLG. PMID- 10789681 TI - Involvement of FK506-sensitive and insensitive granule exocytosis pathways in perforin-dependent target cell lysis mediated by a CD8+ CTL clone. AB - The release of granzyme A and B through granule exocytosis by CD8+ CTL clone OE4 upon T cell receptor (TCR) activation was blocked by FK506 in a dose-dependent manner (IC50 = 3 nM), whereas a significant granzyme release was still detectable even in the presence of excess FK506. In contrast, the production of IFN-gamma was highly sensitive to FK506 (IC50 = 0.01 nM) and could be completely blocked by FK506. Both FK506-sensitive and insensitive granule exocytosis pathways were involved in the actual perforin-dependent killing toward different target cells. The combination of ionomycin and phorbol ester was able to mimic TCR stimulation to induce IFN-gamma production, although the same treatment triggered granule exocytosis inefficiently. Granule exocytosis and IFN-gamma production following TCR activation were profoundly prevented by calphostin C. Thus, these results demonstrate that the granule exocytosis pathway in this CD8+ CTL clone depends on the activation of protein kinase C, and requires either calcineurin-dependent or independent additional signals downstream of TCR activation. PMID- 10789682 TI - T-cell epitope analysis of Mag 3, an important allergen from the house dust mite, Dermatophagoides farinae. AB - Here we describe the detection of T-cell epitope region on the house dust mite allergen Mag 3, which has been shown to trigger T-cell proliferation in mite allergic asthmatic patients. We first examined murine T-cell epitope using T-cell fraction prepared from recombinant Mag 3 (r-Mag 3)-primed H-2k mice. Initial proliferation assay with truncated r-Mag 3 indicated that N-terminal 113 amino acid region was required for triggering T-cell activation. Subsequent epitope scanning with synthetic overlapping peptides revealed that T-cell reactive region was assigned within amino acid range 56-75. We also explored human T-cell determinant using specific T-cells from mite-allergic patients. Intriguingly, we found that amino acid range 56-85, a portion partially overlapping with that identified in r-Mag 3-primed mice, was exclusively recognized by T-cells from different patients. Further investigation of unique T-cell epitope region found in this study would provide insight into the development of animal therapeutic model and/or peptide vaccine for asthma. PMID- 10789684 TI - Activation of gastric afferents increases noradrenaline release in the paraventricular nucleus and plasma oxytocin level. AB - Effects of electrical stimulation of the gastric vagal nerves on plasma levels of oxytocin (OXT) and arginine vasopressin (AVP) were examined in rats anesthetized with urethane. Electrical stimulation of the gastric vagal nerves increased the plasma levels of OXT, but not AVP. The concentrations of extracellular noradrenaline (NA) in the paraventricular nucleus (PVN) were measured by in vivo microdialysis in rats anesthetized with urethane. Electrical stimulation of the gastric vagal nerves evoked an increase followed by a slight decrease in the concentrations of NA. The responses of spontaneous firing magnocellular neurosecretory neurons in the PVN to both electrical stimulation of the gastric vagal nerves and intravenous (i.v.) administration of CCK-8 were examined. Most of the putative OXT-secreting cells recorded were excited by both electrical stimulation of gastric vagal nerves and i.v. administration of CCK-8. These results suggest that gastric vagal afferents activate the central noradrenergic system from the brainstem to the PVN and secretion of OXT. PMID- 10789685 TI - Modulation of the carotid baroreceptor reflex by substance P in the nucleus tractus solitarius. AB - Previous studies have shown that administration of substance P (SP) into the nucleus tractus solitarius (NTS) can evoke a depressor response similar to that produced by activation of the arterial baroreceptors. In addition, some studies have suggested that SP increases the reflex responses to activation of baroreceptor input. The present study was performed to determine the effects of SP on the carotid sinus baroreceptor reflex at the level of the NTS by examining the effects of both exogenous SP microinjected into different rostrocaudal locations in the NTS and blockade of the effects of endogenous SP, through the microinjection of a substance P antagonist (SPa; [D-Pro, D-Trp]-substance P). Changes in pressure in an isolated carotid sinus in anesthetized dogs were used to evoke baroreflex changes in arterial blood pressure (BP) before and after microinjection of SP (0.5 microM) or SPa (10 microM) into barosensitive regions of the NTS. Microinjection of SP or its antagonist did not alter baseline, resting BP but did produce significant changes in baroreflex sensitivity. Microinjection of SP into different rostrocaudal regions of the NTS produced different responses, with rostral and caudal NTS microinjections producing significant increases in sensitivity. No effects on baroreflex sensitivity were obtained in response to SP microinjections into the intermediate NTS. Unlike SP, microinjection of the SPa significantly decreased baroreflex sensitivity at all rostrocaudal levels of the NTS. These data demonstrated that SP has the capability to modulate the carotid baroreflex at the level of the NTS and support a physiological role for endogenously released SP. PMID- 10789683 TI - A peptide based on the CDR3 of an anti-DNA antibody of experimental SLE origin is also a dominant T-cell epitope in (NZBXNZW)F1 lupus-prone mice. AB - A molecular homology has been demonstrated between sequences of the heavy chain variable regions of the anti-DNA, anti-cardiolipin monoclonal antibody, 2C4C2, isolated from C3H.SW mice with induced systemic lupus erythematosus, and sequences of the anti-DNA monoclonal antibody BW16 originating in the lupus-prone (NZBXNZW)F1 mice. It was of interest to determine whether these homologous sequences function also as immunodominant T-cell epitopes, in order to establish a connection between spontaneous and induced experimental models. Therefore, three peptides were designed and synthesized based on the complementarity determining region (CDR)1, CDR2 and CDR3 of the heavy chain of the monoclonal antibody 2C4C2. In the present study, we compare these peptides with the CDR1- and CDR3-based peptides of another murine anti-DNA antibody; namely, 5G12. The comparison was carried out by analyzing the ability of the peptides to induce T cell activation in (NZBXNZW)F1 lupus-prone mice and in mouse strains susceptible to induction of experimental systemic lupus erythematosus. Immunization of (NZBXNZW)F1 mice with the 2C4C2 mAb or with its CDR-based peptides, as well as immunization with the 5G12-based CDR peptides, induced significant lymph node proliferation to the pCDR3 of the 5G12 mAb. Naive (NZBXNZW)F1 splenocytes exhibited activation to the same peptide. It is also shown that MHC class II molecules of (NZBXNZW)F1 macrophages bind preferentially the 5G12-based pCDR3. It is proposed that the CDR3-based peptide of 5G12 mAb of experimental lupus is also a dominant and relevant epitope in the (NZBXNZW)F1 lupus-prone mice. PMID- 10789686 TI - Medial prefrontal depressor response: involvement of the rostral and caudal ventrolateral medulla in the rat. AB - The importance of neurones of the caudal and rostral ventrolateral medulla (CVLM and RVLM, respectively) in mediation of the medial prefrontal cortex depressor response was studied in halothane-anaesthetised rats. Blockade of GABA(A) receptors in the RVLM produced by microinjection of bicuculline (50 nl, 2 mM, n = 6) resulted in reversal of the depressor (-9.5 +/- 1.2 mm Hg) and lumbar sympathetic (-6.5 +/- 5.7 units) responses to pressor (+7.8 +/- 3.5 mm Hg) and sympathoexcitatory (+19.3 +/- 12.5 units) responses and simultaneous blockade of baroreceptor reflex-mediated sympathoinhibition. Baroreflex blockade was reflected by a significant reduction in the gain (slope of the blood pressure vs. lumbar sympathetic nerve discharge regression line) of the reflex. Microinjection of the excitatory amino acid antagonist kynurenic acid (100 nl, 50 mM, n = 6) into the CVLM blocked the baroreflex and significantly reduced the depressor ( 9.6 +/- 0.4 to -6.9 +/- 0.6 mm Hg) and lumbar sympathetic (-4.0 +/- 2.1 to 2.9 +/ 1.9 units) responses to medial prefrontal cortex stimulation. These results support the hypothesis that the medial prefrontal cortex depressor response is mediated by a pathway which converges at the level of the RVLM and which is only partly dependent on an excitatory input to caudal ventrolateral medullary neurones. PMID- 10789687 TI - Local epinephrine release in the rabbit myocardial interstitium in vivo. AB - Although several investigations have suggested cardiac epinephrine (Epi) release, local Epi release in the myocardial interstitium in vivo has not been measured. Using cardiac microdialysis in the rabbit, we measured dialysate Epi and norepinephrine (NE) concentrations as indices of myocardial interstitial Epi and NE levels, respectively. Exocytotic release induced by local administration of KCl (100 mM) through the dialysis probe increased Epi to 24.2 +/- 13.2 pg/ml from a control value of 3.2 +/- 3.6 pg/ml (P < 0.01, n = 6). Non-exocytotic release induced by the local administration of tyramine (10 microg/ml) also increased Epi to 34.6 +/- 15.3 pg/ml (p < 0.05 from control, n = 6). We conclude that Epi can be released via both exocytotic and non-exocytotic release mechanisms from the heart. PMID- 10789688 TI - Modulation of gastrointestinal afferent sensitivity by a novel substituted benzamide (ecabapide). AB - The effects of ecabapide, a novel substituted benzamide compound (3-[2-(3,4 dimethoxyphenyl)ethylcarbamoylmethyl]amino-N-methylb enzamide) that has gastrointestinal prokinetic action, were examined on the discharge of extrinsic afferent nerves supplying the stomach and jejunum in anaesthetized rats. Ecabapide (60 and 180 microg kg(-1), i.v.) had no effect on the baseline discharge of vagal gastric distension-sensitive afferents or the stimulus response profile to gastric distension. Ecabapide also had no effect on either spontaneous jejunal mesenteric afferent nerve discharge or responses to intestinal distension. Ecabapide (180 microg kg(-1)) significantly inhibited the maximum discharge of jejunal afferents induced by cholecystokinin (CCK8; 50 pmol, i.v.), whereas it failed to inhibit the excitatory action of 2-methyl-5 hydroxytryptamine (2Me-5-HT; 10 microg, i.v.), a selective 5-HT3 receptor agonist. A model of acute focal intestinal ischaemia was used to evaluate the actions of ecabapide on the discharge of activated jejunal afferents. Ischaemia produced a substantial increase in afferent discharge which was reproducible when the duration of ischaemia was limited to less than 10 min and repeated every 15 min. Ecabapide at doses of 60 and 180 microg kg(-1) significantly reduced ischaemia-induced increases in afferent discharge. In addition to its therapeutic efficacy as a gastrointestinal prokinetic agent, these findings show also that ecabapide may also have an inhibitory action on the discharge of intestinal afferents activated by ischaemia. PMID- 10789689 TI - Immunolocalisation of myosin-V in the enteric nervous system of the rat. AB - We show here the localisation of myosin-V in whole mount preparations of the mucous-submucous and the muscular layers of rat small intestine by using an affinity purified antibody specific to the tail domain of myosin-V. Myosin-V immunostaining was intense in the submucous and myenteric nervous plexuses, allowing the visualisation of neuronal cell bodies and fibres. Western blots of total muscle layers homogenates detected with the same antibody revealed a single band of the expected size for myosin-V. Understanding the cellular localisation and function of this class of myosin is an important challenge and the accessibility and simplicity of the enteric nervous system as compared to the central nervous system, makes the digestive tract an attractive model for studying possible functional roles of myosin-V in neurotransmission and neuroplasticity. PMID- 10789690 TI - Effect of unilateral superior cervical ganglionectomy on bone mineral content and density of rat's mandible. AB - To assess the effect of a local sympathectomy on bone metabolism, the effect of a unilateral superior cervical ganglionectomy (Gx) on growth and bone mineral content and density of the ipsi- and contralateral mandibles was examined in female rats. A significant increase in the hemi-mandibular bone ipsilateral to Gx was found as compared to the contralateral, sham-operated side 30 days, but not 15 days, after surgery. Bone mineral content of the hemi-mandibular bones was significantly lower in the side ipsilateral to Gx in the group of rats killed on the 30th day after surgery. Since no difference in areas between innervated and denervated hemi-mandibles was found, bone mineral density was also significantly lower in the hemi-mandible ipsilateral to Gx. The results further support that a regional sympathectomy causes qualitative alterations in bone modeling and remodeling, leading to bone resorption. PMID- 10789691 TI - Direct interaction between the sympathetic and renin-angiotensin system in myocardial tissue: a microdialysis study in anaesthetised rats. AB - It has been suggested that local activation of the renin-angiotensin system is involved in early stages of myocardial pathophysiology. To date, there is increasing evidence for interactions between the renin-angiotensin system and the sympathetic nervous system; consequently, local sympathetic activation may also be involved in this. Microdialysis has great potential in the direct investigation of neurohormonal interactions. Therefore, the present study employs microdialysis to study the local effects of exogenous angiotensin II on the interstitial norepinephrine concentration of the normally innervated left ventricle of the anaesthetised rat. The present study investigates the effect of increasing dosages of exogenous angiotensin II on local interstitial norepinephrine. Furthermore, a single dose of losartan was infused on top of the highest dose of angiotensin II, in order to study possible involvement of angiotensin II type 1 (AT1) receptors. Both infusion and sampling were carried out locally, via the microdialysis probes. Concomitantly, circulating norepinephrine levels, heart rate and respiratory rate were monitored to evaluate physiologic stability of the preparation throughout the experiment. Time controls consisted of rats that were perfused with only a Ringer's solution. Angiotensin II induced a dose dependent increase in norepinephrine that was significantly reduced by losartan. Norepinephrine levels in both plasma (infusion experiment and time controls) and the left ventricular wall (time controls) remained stable throughout the experiment, just as heart rate and respiratory rate did. This study for the first time employs microdialysis to demonstrate direct interaction between the sympathetic nervous system and the renin-angiotensin system in the rat left ventricle. The data strongly suggest that AT1 receptors are involved in this interaction, since selective AT1 receptor blockade with losartan significantly reduced the angiotensin II induced norepinephrine concentration. PMID- 10789692 TI - GABA(A) receptors on calbindin-immunoreactive myenteric neurons of guinea pig intestine. AB - These studies were carried out to characterize the properties of gamma aminobutyric acidA (GABA(A)) receptors on guinea pig intestinal myenteric neurons maintained in primary culture. In addition, the type of neuron expressing GABA(A) receptors was identified using immunohistochemical methods. Whole-cell patch clamp recordings of currents elicited by GABA and acetylcholine (ACh) were obtained using pipettes containing Neurobiotin. After electrophysiological studies, neurons were processed for localization of calbindin-D28K immunoreactivity (calbindin-ir). GABA (1 mM) and ACh (3 mM) caused inward currents in most cells tested. GABA currents were mimicked by muscimol (1-300 microM) and were blocked by bicuculline (10 microM) indicating that GABA was acting at GABA(A) receptors. GABA currents were associated with a conductance increase and a linear current/voltage relationship with a reversal potential of 1 +/- 1 mV (n = 5). Pentobarbital (PB, 3-1000 microM) and diazepam (DZP, 0.01-10 microM) potentiated GABA-induced currents. A maximum concentration of DZP (1 microM) increased GABA-induced currents 3.1 +/- 0.3 times while PB (1000 microM) increased GABA currents by 11 +/- 2 times. In outside-out patches, the amplitude of GABA-activated single-channel currents was linearly related to membrane potential with a single-channel conductance of 28.5 + 0.5 pS (n = 10). PB and DZP increased the open probability of GABA-induced single-channel currents. Neurons containing calbindin-ir were large, were isolated from other neurons and had GABA current amplitudes of -3.4 +/- 0.3 nA (n = 48). Neurons with weak or absent calbindin-ir were smaller, were localized in clusters of cells and had GABA induced current amplitudes of -0.6 +/- 0.1 nA (n = 20). ACh-induced currents were smaller in calbindin-ir neurons (-0.7 +/- 0.1 nA) compared to weakly calbindin-ir neurons (-1.4 +/- 0.1 nA). These results indicate that myenteric calbindin-ir neurons express a high density of GABA(A) receptors. Cell size and location allow visual identification of neurons likely to contain calbindin-ir permitting targeted studies of the properties of these neurons. PMID- 10789693 TI - Evaluation of direct and axon reflex sweating in the mouse. AB - The nicotinic axon reflex-mediated sudomotor response was studied in mice and rats by recording the impressions of sweat droplets made in silicone molds after local injection of nicotine, and compared with sweating induced by acetylcholine and pilocarpine. Nicotine failed to activate mouse plantar sweat glands at any of the concentrations used (from 3 x 10(-6) to 3 x 10(-1) M). On the contrary, both acetylcholine and pilocarpine produced a dose-dependent increase in the number of secreting sweat glands. The location of sweat glands reactive to pilocarpine and acetylcholine was similar and restricted to the pads near the site of injection. We conclude that the sudomotor axon reflex response mediated by nicotinic receptors is not present in the mouse and the rat. PMID- 10789694 TI - Phase transitions in the common brainstem and related systems investigated by nonstationary time series analysis. AB - Neuronal activities of the reticular formation (RF) of the lower brainstem and the nucleus tractus solitarii (NTS, first relay station of baroreceptor afferents) were recorded together in the anesthized dog with related parameters of EEG, respiration and cardiovascular system. The RF neurons are part of the common brainstem system (CBS) which participates in regulation and coordination of cardiovascular, respiratory, somatomotor systems, and vigilance. Multiple time series of these physiological subsystems yield useful information about internal dynamic coordination of the organism. Essential problems are nonlinearity and instationarity of the signals, due to the dynamic complexity of the systems. Several time-resolving methods are presented to describe nonlinear dynamic couplings in the time course, particularly during phase transitions. The methods are applied to the recorded signals representing the complex couplings of the physiological subsystems. Phase transitions in these systems are detected by recurrence plots of the instationary signals. The pointwise transinformation and the pointwise conditional coupling divergence are measures of the mutual interaction of the subsystems in the state space. If the signals show marked rhythms, instantaneous frequencies and their shiftings are demonstrated by time frequency distributions, and instantaneous phase differences show couplings of oscillating subsystems. Transient signal components are reconstructed by wavelet packet time selective transient reconstruction. These methods are useful means for analyzing coupling characteristics of the complex physiological system, and detailed analyses of internal dynamic coordination of subsystems become possible. During phase transitions of the functional organization (a) the rhythms of the central neuronal activities and the peripheral systems are altered, (b) changes in the coupling between CBS neurons and cardiovascular signals, respiration and the EEG, and (c) between NTS neurons (influenced by baroreceptor afferents) and CBS neurons occur, and (d) the processing of baroreceptor input at the NTS neurons changes. The results of this complex analysis, which could not be done formerly in this manner, confirm and complete former investigations on the dynamic organization of the CBS with its changing relations to peripheral and other central nervous subsystems. PMID- 10789695 TI - Potassium channel blockade induces action potential generation in guinea-pig airway vagal afferent neurones. AB - Electrophysiological studies of vagal sensory nerves with cell bodies in the nodose ganglion and mechanically sensitive receptive fields in the guinea-pig trachea/bronchus, were performed. Exposure of the mechanically sensitive receptive fields to 4-aminopyridine (100 microM-1 mM) caused pronounced action potential discharge in all fibres studied. Action potential generation was also produced by alpha-dendrotoxin, and in a subset of fibres, by barium. By contrast, neither iberiotoxin, tetraethyl ammonium, glybenclamide, BDS-II, nor apamin caused action potential generation in the vagal afferent nerve fibres. Tetramethylrhodamine dextran was instilled into the trachea to retrogradely label cell bodies within the nodose ganglion. In these cells, 4-aminopyridine caused a large depolarization of the resting membrane potential, concomitant with an increase in input impedance. The data suggest 4-aminopyridine- and alpha dendrotoxin-sensitive ion channels within the airway afferent nerve membrane hold the resting membrane potential below the threshold for action potential generation. Mechanisms that lead to an inhibition of these channels will likely lead to an increase in excitability of the airway afferent neurones. PMID- 10789696 TI - Interaction between AVP and sympathetic system in subtotal nephrectomy-saline hypertension: role of alpha and V1 receptors. AB - The development process of subtotal nephrectomy-salt hypertension is still unclear. The aim of the present study was to determine the role of the interaction between vasopressin and sympathetic system in the development of this hypertension by using AVP V1 antagonist and alpha blocker phentolamine under anesthesia condition. For this purpose, we carried out about 73% subtotal nephrectomy on male Wistar rats. One group of these rats (normotensive group) was given a low-salt diet and the other group (hypertensive group) was given a high salt diet for 4 weeks. Finally, eight groups of rats were formed according to the kind(s) of the injected drug(s): (1) normotensive and hypertensive groups injected only V1 antagonist, (2) normotensive and hypertensive groups injected only phentolamine, (3) normotensive and hypertensive groups injected first V1 antagonist and then phentolamine, (4) normotensive and hypertensive groups injected first phentolamine and then V1 antagonist. Either V1 or alpha blockage separately led to a higher reduction in the mean blood pressure (MAP) of the hypertensives than, of the normotensives (p < 0.05). The combined blockage of V1 and alpha receptors, also caused a higher decrease in the MAP of hypertensive group than, of normotensive group, not depending on the order of the injections (p < 0.01). The heart rate increase recorded as a response to the phentolamine injection in normotensive group, did not develop in hypertensive group (p < 0.05). There was no significant difference between the two groups with regard to plasma electrolytes and osmolality. A positive correlation was found between systolic blood pressure and plasma osmolality in hypertensive group (r = 0.40, p < 0.05), but not in normotensive group. We conclude that the increase in V1 and alpha pressor activities contributes to the subtotal nephrectomy-saline hypertension and the augmentation of alpha pressor activity by vasopressin may participate in this contribution. PMID- 10789697 TI - Augmentation of somato-sympathetic reflex in the ischemic hindlimb of anesthetized rats. AB - Activities of the visceral sympathetic nerve increase with noxious mechanical stimulation. This study investigated the effects of noxious mechanical stimulation of hindlimb on arterial blood pressure, heart rate (HR) and renal sympathetic nerve activities (RNA) in anesthetized rats intact or under ischemic conditions. This study utilized two methods of noxious mechanical stimulation. One was pinching of the hindpaw; the other, strangulation of the ankle joint. Twenty-three male Wistar rats were used in the experiment. Pinching and strangulation of the intact hindlimb caused an increase in RNA. The increased RNA was greater during strangulation than during pinch stimulation (p < 0.0001). In the ischemic hindlimb produced by occlusion of the left common iliac artery, pinching and strangulation caused a further increase in RNA (p < 0.0001). Arterial blood pressure and HR significantly increased during strangulation with ischemic condition (p < 0.05). These results suggest that the reflex effects of somatosensory input on RNA were augmented in the ischemic hindlimb. PMID- 10789698 TI - Glutamatergic synaptic inputs activate neurons in the subfornical organ through non-NMDA receptors. AB - The subfornical organ (SFO) plays an important role in central regulation of the autonomic nervous system. The synaptic transmission properties of neurons in the SFO were studied with intracellular and whole-cell patch clamp recordings in the rat slice preparations. Both the spontaneous and evoked excitatory postsynaptic potentials (EPSPs) and currents (EPSCs) were almost completely suppressed by the glutamate receptor antagonist kynurenic acid and the non-NMDA (N-methyl-D aspartic acid) antagonist CNQX. The non-NMDA agonist kainic acid depolarized the membrane most potently, compared with NMDA and quisqualic acid. These suggest that glutamate is a main excitatory neurotransmitter in the SFO and that its action is at least partly mediated through non-NMDA receptors. PMID- 10789699 TI - Surveillance for foodborne-disease outbreaks--United States, 1993-1997. AB - PROBLEM/CONDITION: Since 1973, CDC has maintained a collaborative surveillance program for collection and periodic reporting of data on the occurrence and causes of foodborne-disease outbreaks (FBDOs) in the United States. REPORTING PERIOD COVERED: This summary reviews data from January 1993 through December 1997. DESCRIPTION OF SYSTEM: The Foodborne-Disease Outbreak Surveillance System reviews data concerning FBDOs, defined as the occurrence of two or more cases of a similar illness resulting from the ingestion of a common food. State and local public health departments have primary responsibility for identifying and investigating FBDOs. State, local, and territorial health departments use a standard form to report these outbreaks to CDC. RESULTS: During 1993-1997, a total of 2,751 outbreaks of foodborne disease were reported (489 in 1993, 653 in 1994, 628 in 1995, 477 in 1996, and 504 in 1997). These outbreaks caused a reported 86,058 persons to become ill. Among outbreaks for which the etiology was determined, bacterial pathogens caused the largest percentage of outbreaks (75%) and the largest percentage of cases (86%). Salmonella serotype Enteritidis accounted for the largest number of outbreaks, cases, and deaths; most of these outbreaks were attributed to eating eggs. Chemical agents caused 17% of outbreaks and 1% of cases; viruses, 6% of outbreaks and 8% of cases; and parasites, 2% of outbreaks and 5% of cases. INTERPRETATION: The annual number of FBDOs reported to CDC did not change substantially during this period or from previous years. During this reporting period, S. Enteritidis continued to be a major cause of illness and death. In addition, multistate outbreaks caused by contaminated produce and outbreaks caused by Escherichia coli O157:H7 remained prominent. ACTIONS TAKEN: Current methods to detect FBDOs are improving, and several changes to improve the ease and timeliness of reporting FBDO data are occurring (e.g., a revised form to simplify FBDO reporting by state health departments and electronic reporting methods). State and local health departments continue to investigate and report FBDOs as part of efforts to better understand and define the epidemiology of foodborne disease in the United States. At the regional and national levels, surveillance data provide an indication of the etiologic agents, vehicles of transmission, and contributing factors associated with FBDOs and help direct public health actions to reduce illness and death caused by FBDOs. PMID- 10789700 TI - Endoscopic saphenous vein harvest decreases leg wound complication in coronary artery bypass grafting patients. AB - BACKGROUND: Coronary artery bypass grafting (CABG) is the most common procedure performed in adult cardiovascular surgery. The most frequently used conduit is the greater saphenous vein. Using traditional methods, the complication rate of the leg is relatively high (up to 24%). To decrease the complication rate, we used the Endo-Path to harvest the greater saphenous vein. METHODS AND RESULTS: From May 1997 through March 1999, a total of 135 patients received the CABG operation. We excluded the patients who died immediately postoperatively or had concomitant surgical procedures. Sixty patients received the endoscopic saphenous vein harvest procedure (group A), while another 59 patients (group B) did not. No important differences were noted between the two groups in respect to the number of distal anastomoses, length of harvested vein, total surgical time, and length of ICU stay. However, the leg wound complication rate decreased from 20.3% to 5.0% (p < 0.001). CONCLUSIONS: Although the long-term patency rate needs time to be proven, the endoscopic greater saphenous vein harvest method is an attractive and effective method. PMID- 10789701 TI - Acute mechanical valve thrombosis of the St. Jude medical prosthesis. AB - From 1986 to 1996, 2585 patients underwent valve replacement with the St. Jude medical prosthesis. Sixty experienced mechanical valve thrombosis. Seventeen of 60 patients (28.3%) had isolated aortic valve replacements, 33 had isolated mitral valve replacements (55%), and 10 had double valve replacements (16.7%) (aortic and mitral valve replacement). All patients who underwent reoperation for mechanical valve thrombosis were functional Class III or IV. Against medical advice, systemic anticoagulation with warfarin sodium had been discontinued or used only intermittently. Thus, anticoagulant activity was not adequate. The diagnosis of thrombosis was made by clinical examination, laboratory findings, and echocardiography and cineradiography. Of the 60 patients, 9 patients died early after surgery or before discharge. Most of the deaths were attributed to low cardiac output. The overall hospital mortality was 15%. The overall 10-year actuarial survival rate was 82.8+/-1.6%. In our study, reoperation for thrombosed mechanical prosthesis was not an independent parameter determining mortality. Age was the only statistically important hospital mortality predictor. Of this group, 90% suffered mechanical valve obstruction within the first 5 years after operation. These results suggest that valve re-replacement appears to be a suitable surgical treatment for thrombosis of mechanical prosthetic valves, especially in the young. In these patients subsequent anticoagulation management is necessary. PMID- 10789702 TI - Intrapericardial teratoma in a newborn: a case report. AB - Intrapericardial teratomas are unusual tumors that often cause respiratory distress and might be lethal in the newborn. The purpose of this article is to present the clinical and pathological findings in a 12-day-old male successfully treated for a big intrapericardial teratoma. Given that the diameter of this tumor is generally related to the age at the time of diagnosis, the rarity of our case is the presence of a huge intrapericardial teratoma in a newborn. The surgical resection was lifesaving. PMID- 10789703 TI - Predictive value of preoperative serum cholinesterase concentration in patients with liver dysfunction undergoing cardiac surgery. AB - OBJECTIVE: There are an increasing number of patients with severe liver dysfunction subjected to open heart surgery. This retrospective study was designed to assess operative results and clarify the degree of liver injury in patients with liver dysfunction undergoing open heart surgery. In addition, determinants influencing their prognosis were assessed. METHODS: In a 9-year period from 1988 to 1996, we operated on 31 patients with posthepatitis liver dysfunction and 16 with chronic passive congestion of the liver. This group was 2.3% and 1.6% of the 1368 patients undergoing cardiac surgery in the same period. We compared several perioperative factors between survivors and nonsurvivors to determine risk factors affecting mortality. RESULTS: In the group with posthepatitis liver dysfunction, the postoperative course of 5 patients among 31 (16.1%) was poor. Serum cholinesterase concentration was lower only in the nonsurvivor group (nonsurvivor vs survivor: 1979+/-949 vs 3515+/-1424 IU/I, p < 0.05). All patients with cholinesterase < 2000 IU/L died. The duration of CPB (212+/-53 vs 150+/-54 minutes, p < 0.03) and ACC time (151+/-38 vs 96+/-40 minutes, p < 0.02) was longer in the nonsurvivor group. In the group with chronic passive congestion, the postoperative course of 5 of 16 (31.3%) patients with valvular disease was poor. Serum cholinesterase concentration was lower only in the nonsurvivor group (nonsurvivor vs survivors: 2006+/-435 vs 3483+/-1442 IU/L, p < 0.02), and all patients with cholinesterase < 2000 IU/L died. Postoperative bleeding was greater in the nonsurvivor group (3327+/-2106 vs 1428+/-643 mL, p < 0.05). Multivariate logistic regression analysis including the described pre- and intraoperative factors identified only serum cholinesterase concentration (F = 9.18) as significant. CONCLUSIONS: A low value of preoperative serum cholinesterase (< 2,000 IU/L) is thought to be the predictor of prognosis after open heart surgery in patients with severe posthepatitis and congestive liver dysfunction. Operative factors (cardiopulmonary time in posthepatitis liver dysfunction and postoperative bleeding in the congestive liver dysfunction) also influenced the prognosis. PMID- 10789704 TI - Idiopathic saccular aneurysm of the azygos vein. AB - Idiopathic saccular aneurysm of the azygos vein is a rare condition. We report the case of a 52-year-old man with saccular aneurysm of the azygos vein who underwent surgical resection. Preoperative dynamic computed tomography revealed strong and rapid enhancement of the chest mass. Magnetic resonance imaging showed a thoracic tumor with low signal intensity on the T1-weighted image and coexistence of low and high signal intensity on the T2-weighted image. Intraoperative findings showed cystic dilatation of the azygos vein. Although an accurate preoperative diagnosis of saccular azygos vein aneurysm is difficult, especially differential diagnosis between this anomaly and mediastinal tumors, a markedly enhanced mass shown by dynamic computed tomography was useful for the preoperative diagnosis of this anomaly. PMID- 10789705 TI - Quantitative digital angiography as an adjunct to the intraoperative placement of endovascular stents in congenital heart disease. AB - Clearly identifiable intraoperative landmarks render the placement of intraoperative stents difficult. Preoperative use of quantitative digital angiography helps the surgeon accurately insert endovascular stents intraoperatively. By using defined points of reference, we were able to carefully select the size and lengths of stents before the operation and precisely place these stents in the operating room. Furthermore, we have been able to redilate these stents using the same techniques at subsequent operations. Our results reflect the efficacy of this technique. PMID- 10789706 TI - Reoperative median sternotomy using a cast spreader. AB - Repeat median sternotomy carries a high mortality rate secondary to a higher incidence of injury to the underlying vital structures. The reported incidence of reentry accidents may be as high as 6% to 10%. We describe a new technique of redo sternotomy using a nitrogen-powered oscillating saw and a cast spreader. The new technique was used for 89 consecutive cases without any incidence of injury to the underlying structures. The use of a case spreader during repeat median sternotomy may enhance the safety of reentry. PMID- 10789707 TI - Protective effect of JTV519, a new 1,4-benzothiazepine derivative, on prolonged myocardial preservation. AB - BACKGROUND: JTV519 is know to protect cardiomyocytes from calcium overloading induced damage. The aim of this study was to investigate the potential protective effect of JTV519 on myocardium subjected to prolonged ischemia and the underlying mechanism of such protection. The effect of JTV519 was also compared with that of diltiazem, a 1,5-benzothiazepine derivative. METHODS: Isolated rat hearts were randomly divided into three groups. Control hearts were arrested with histidine tryptophan-ketoglutarat (HTK) cardioplegic solution alone. In the JTV519 group of hearts, cardiac arrest was achieved with JTV519 (10(-3) mmol/L) in the HTK solution. Hearts in the diltiazem group were arrested with diltiazem (0.5 mmol/L) in the HTK solution. All the hearts were then subjected to 6-hour storage in HTK solution at 4 degrees C. RESULTS: After a 30-minute reperfusion, the left ventricular developed pressure in the JTV519 and diltiazem groups were improved significantly compared with the control group. There was a significantly lower left ventricular end-diastolic pressure level and higher recovery of coronary flow in the JTV519 group than in the control group. The postischemic intracellular calcium concentration was attenuated by adding JTV519 or diltiazem to HTK cardioplegia. CONCLUSION: As an adjunct to cardioplegia, JTV519 showed a significant protective effect on myocardium undergoing 6 hours of ischemia. The beneficial protective effects of JTV519 are correlated with its ability to inhibit the postischemic rise in intracellular calcium. PMID- 10789708 TI - Proximal aortic perfusion with passive cerebral flow: a method to prevent cerebral embolism in the arch and descending aortic operation. AB - Antegrade aortic perfusion is preferable in arch and descending aortic operations. We describe a left lateral approach using an ascending aortic cannula, temporarily relocated within the distal aorta to maintain hypothermic cardiopulmonary bypass of the lower torso. This modification provides continual antegrade systemic perfusion and passive retrograde cerebral flow during arch repair. It minimizes the risk of embolization into the brachiocephalic arteries of debris and malperfusion of the dissected aorta. PMID- 10789709 TI - New horizons on the surgical treatment of dilated cardiomyopathy. PMID- 10789710 TI - Left ventricular volume reduction and reconstruction in ischemic cardiomyopathy. AB - BACKGROUND: Ischemic cardiomyopathy can be the result of large or small myocardial infarctions or due to myocardial hibernation. Patients with an end systolic volume index >100 mL?m2 do not benefit from revascularization alone and require an operation that reduces ventricular volume. Various approaches to reduce ventricular volume have been described. We applied several of these techniques in patients with end-stage ischemic cardiomyopathy. METHODS: Forty eight patients with end-stage ischemic cardiomyopathy (Class III-IV) underwent left ventricular volume reduction operations with coronary revascularization and mitral valve repair or Alfieri valvoplasty. Fourteen patients underwent interpapillary resections, 22 anterior resections, 4 posterior resections, 2 anterior and posterior resections, and 6 patients reduction of left ventricular volume with endocavitary patches. RESULTS: All the techniques used improved left ventricular function. Analysis of mortality revealed that extensive resections (interpapillary, anterior, and posterior resection) had a 43% mortality. However, a limited resection or a ventricular reconstruction with an endocavitary patch had only a 12.5% mortality. When we changed our approach to a more conservative one, mortality was reduced from 26% the first 12 months to 13% in the last 15 months of the study. CONCLUSIONS: Ischemic cardiomyopathy has a poor prognosis if the end-systolic volume index exceeds 100 mL/m2. Various procedures exist to reduce left ventricular volume. Extensive ventricular resections improve ventricular function, but have a high mortality. This led us to use other methods of ventricular volume reduction such as more conservative resections combined with left ventricular reconstructions or ventricular volume reduction with endocavitary patches. Mortality was reduced significantly by this approach. The patients that survived have remained Class I-II in a follow-up that extends up to 30 months. Surgical therapy of Class III-IV ischemic cardiomyopathy is feasible, but aggressive ventricular resections have a high mortality. We advocate a more reconstructive approach with limited or no ventricular resection. PMID- 10789711 TI - Partial left ventriculectomy (Batista procedure) in the treatment of dilated cardiomyopathy: Makati Medical Center Philippine experience. AB - In the Philippines patients with end-stage heart disease refractory to conventional medical and surgical treatment do not have alternative choices. More than 99% of the population cannot afford cardiac transplantation. Partial left ventriculectomy (PLV) is a surgical procedure that improves cardiac function and refractory congestive heart failure (CHF). Between October 1997 and February 1998 eight patients had PLV at the Makati Medical Center, Philippines. All patients had end-stage dilated cardiomyopathy. Six patients had an idiopathic etiology, one was ischemic and one valvular. Seven of eight operations were done with the heart beating and all had transesophageal echo monitoring. An average of a 2-cm reduction in the left ventricle diameter was achieved and ejection fraction improved in all cases. There were no operative deaths. There were three late deaths. Two patients died of refractory CHF and ventricular arrhythmias and one patient died of massive cerebral hemorrhage with coumadin therapy. The five survivors are all doing well with no CHF. Follow-up two-dimensional echo shows stable left ventricular (LV) size and improved ejection fraction. Our initial experience shows that PLV, at least in the short-term, has beneficial effects in the treatment of end-stage dilated cardiomyopathy and might become an alternative to cardiac transplantation. PMID- 10789713 TI - Mechanical circulatory devices for the treatment of heart failure. AB - BACKGROUND: During the last four decades substantial efforts have been made in the development of effective mechanical circulatory devices. Since the first clinical utilization in the 1960s, the field has gone from the stage of clinical experimentation to that of a valid and effective heart failure treatment alternative. Experience gained during the short-term use of these devices, typically as a bridge to cardiac transplantation, has led to increased expectations of devices capable of long-term or permanent support to be used as a permanent treatment for end-stage heart failure patients. This article reviews the history, current state of the art, and future of the field of mechanical circulatory devices. METHODS: Mechanical circulatory devices can be classified into three major categories: (1) total artificial hearts, (2) pulsatile ventricular assist devices, and (3) nonpulsatile ventricular assist devices. The most widely used devices have been the pulsatile ventricular assist devices with more than 5,800 reported cases, whereas the use of total artificial hearts has been limited to less than 350 reported cases. Nonpulsatile devices have been used clinically, but only in short-term cases (i.e., hours and days), whereas the pulsatile devices have been used in the long-term application, with patients supported for weeks, months, and in a small number of cases, years. The technological evolution of these devices has gone from large, extracorporeal systems designed to keep the patient alive in the intensive care unit (ICU) until a donor organ could be found, to portable devices that allow the patient to be mobilized outside of the hospital setting. RESULTS: The clinical experience with mechanical circulatory devices as a bridge to cardiac transplantation has saved the lives of thousands of patients. Exciting new research discoveries related to recovery of native heart function during extended circulatory support have provided new hope for many more patients. Additional research efforts currently underway are being tested at various laboratories around the world and will soon provide the next generation of systems. These new systems will offer the recipients an unparalleled quality of life with minimal limitations on daily activities. The progress in this field has reached the point where circulatory support will soon be considered a valid long-term or permanent therapy and an elective to transplantation. PMID- 10789714 TI - The future of the surgical treatment of dilated cardiomyopathy. PMID- 10789712 TI - Intermediate-term results of partial left ventriculectomy. AB - Partial left ventriculectomy (PLV) has been introduced as an option for patients with end-stage dilated cardiomyopathy. We report the result of a prospective trial of PLV in patients with idiopathic dilated cardiomyopathy, left ventricular end-diastolic diameter (LVEDD) > 7 cm, refractory New York Heart Association (NYHA) Class IV symptoms, and depressed exercise oxygen consumption studies. Sixteen patients underwent a PLV with a mean follow-up of 13 months. Fourteen patients were male. Mean age was 49.6+/-10.5 years (range 30 to 67 years). Left ventricular ejection fraction (LVEF) improved after surgery from 13.9+/-5.6 to 21.0+/-8.4, and this improvement persisted for up to 12 months after operation. LVEDD and NYHA Class also were significantly improved after surgery and for up to 12 months of follow-up. Operative mortality was 6.25%. Twelve-month survival was 86% by Kaplan-Meier analysis. Four (25%) of 16 patients that had initial improvement after PLV developed recurrent heart failure and were listed for transplantation. Freedom from need for listing for heart transplantation was 65% at 12 months. Freedom from death or the need for relisting at 12 months was 56%. PLV can be performed with acceptable early and 12-month mortality. Significant improvements in LVEF, LVEDD, and NYHA Class are seen at up to 12 months of follow up. Some patients will develop recurrent heart failure and require relisting for heart transplantation. PMID- 10789715 TI - UKCCCR guidelines for the use of cell lines in cancer research. PMID- 10789716 TI - Human oesophageal adenocarcinoma cell lines JROECL 47 and JROECL 50 are admixtures of the human colon carcinoma cell line HCT 116. AB - In two recently described human oesophageal adenocarcinoma cell lines JROECL 47 and JROECL 50, derived from one tumour, we detected identical E-cadherin and beta catenin gene mutations as in colon carcinoma cell line HCT 116. We demonstrate by HLA-typing, mutation analysis and microsatellite analysis that cell lines JROECL 47 and JROECL 50 are admixtures of the human colon adenocarcinoma cell line HCT 116. PMID- 10789718 TI - Clinical pharmacokinetic and in vitro combination studies of nolatrexed dihydrochloride (AG337, Thymitaq) and paclitaxel. AB - A clinical study of nolatrexed dihydrochloride (AG337, Thymitaq) in combination with paclitaxel was performed. The aims were to optimize the schedule of administration and determine any pharmacokinetic (PK) interactions between the two drugs. In vitro combination studies were performed to assist with schedule optimization. Three patients were entered on each of three different schedules of administration of the two drugs: (1) paclitaxel 0-3 h, nolatrexed 24-144 h; (2) nolatrexed 0-120 h, paclitaxel 48-51 h; (3) nolatrexed 0-120 h, paclitaxel 126 129 h. Paclitaxel was administered at a dose of 80 mg m(-2) over 3 h and nolatrexed at a dose of 500 mg m(-2) day(-1) as a 120-h continuous intravenous infusion. Plasma concentrations of both drugs were determined by high performance liquid chromatography. In vitro growth inhibition studies using corresponding schedules were performed using two head and neck cancer cell lines. In both HNX14C and HNX22B cell lines, synergistic growth inhibition was observed on schedule 2, whereas schedules 1 and 3 demonstrated antagonistic effects. In the clinical study, there was no effect of schedule on the pharmacokinetics of nolatrexed. However, patients on schedules 1 and 3 had a higher clearance of paclitaxel (322-520 ml min(-1) m(-2)) than those on schedule 2 (165-238 ml min( 1) m(-2)). Peak plasma concentrations (1.66-1.93 vs. 0.86-1.32 microM) and areas under the curve (392-565 vs. 180-291 microM min(-1)) of paclitaxel were correspondingly higher on schedule 2. The pharmacokinetic interaction was confirmed by studies with human liver microsomes, nolatrexed being an inhibitor of the major routes of metabolism of paclitaxel. Toxicity was not schedule dependent. Nolatrexed and paclitaxel may be safely given together when administered sequentially at the doses used in this study. Studies in vitro suggest some synergy, however, due to a pharmacokinetic interaction, paclitaxel doses should be reduced when administered during nolatrexed infusion. PMID- 10789717 TI - Vascular permeability in a human tumour xenograft: molecular charge dependence. AB - Molecular charge is one of the main determinants of transvascular transport. There are, however, no data available on the effect of molecular charge on microvascular permeability of macromolecules in solid tumours. To this end, we measured tumour microvascular permeability to different proteins having similar size but different charge. Measurements were performed in the human colon adenocarcinoma LS174T transplanted in transparent dorsal skinfold chambers in severe combined immunodeficient (SCID) mice. Bovine serum albumin (BSA) and IgG were fluorescently labelled and were either cationized by conjugation with hexamethylenediamine or anionized by succinylation. The molecules were injected i.v. and the fluorescence in tumour tissue was quantified by intravital fluorescence microscopy. The fluorescence intensity and pharmacokinetic data were used to calculate the microvascular permeability. We found that tumour vascular permeability of cationized BSA (pI-range: 8.6-9.1) and IgG (pI: 8.6-9.3) was more than two-fold higher (4.25 and 4.65x10(-7) cm s(-1)) than that of the anionized BSA (pI approximately 2.0) and IgG (pI: 3.0-3.9; 1.11 and 1.93x10(-7) cm s(-1), respectively). Our results indicate that positively charged molecules extravasate faster in solid tumours compared to the similar-sized compounds with neutral or negative charges. However, the plasma clearance of cationic molecules was approximately 2x faster than that of anionic ones, indicating that the modification of proteins enhances drug delivery to normal organs as well. Therefore, caution should be exercised when such a strategy is used to improve drug and gene delivery to solid tumours. PMID- 10789719 TI - The constitutive level of vascular endothelial growth factor (VEGF) is more important than hypoxia-induced VEGF up-regulation in the angiogenesis of human melanoma xenografts. AB - Angiogenesis of tumours might develop as a result of environmental conditions, such as hypoxia, and/or as a result of genetic alterations specific for tumour cells. The relative contributions of these mechanisms were investigated by comparing the in vivo expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) to the hypoxic fraction, the angiogenic potential and the vascular density of four human melanoma lines (A-07, D-12, R 18, U-25) grown intradermally in Balb/c nu/nu mice. VEGF expression, bFGF expression and expression of pimonidazole, a marker of hypoxic cells, were investigated by immunohistochemistry. An association between high VEGF and bFGF expression and high angiogenic potential was detected, suggesting an important role for VEGF/bFGF in the angiogenesis of melanomas. High VEGF/bFGF expression was also related to low hypoxic fraction and high vascular density. Thus, the constitutive, genetically determined level of VEGF was probably more important than hypoxia-induced upregulation in the angiogenesis of the melanoma xenografts. PMID- 10789720 TI - Role of tumour markers in monitoring epithelial ovarian cancer. AB - Currently the only tumour marker to have a well-defined and validated role in the management of ovarian cancer is CA125. Changes in the level of CA125 can be used as a reliable indication of response or progression according to various criteria, but it does not yet have a clear place in diagnosis or prognosis. Its value as part of a screening tool and during routine follow-up remain the subject of ongoing trials. Other markers remain experimental and do not have a well defined contribution to make at present. PMID- 10789721 TI - Increased local cytostatic drug exposure by isolated hepatic perfusion: a phase I clinical and pharmacologic evaluation of treatment with high dose melphalan in patients with colorectal cancer confined to the liver. AB - A phase I dose-escalation study was performed to determine whether isolated hepatic perfusion (IHP) with melphalan (L-PAM) allows exposure of the liver to much higher drug concentrations than clinically achievable after systemic administration and leads to higher tumour concentrations of L-PAM. Twenty-four patients with colorectal cancer confined to the liver were treated with L-PAM dosages escalating from 0.5 to 4.0 mg kg(-1). During all IHP procedures, leakage of perfusate was monitored. Duration of IHP was aimed at 60 min, but was shortened in eight cases as a result of leakage from the isolated circuit. From these, three patients developed WHO grade 3-4 leukopenia and two patients died due to sepsis. A reversible elevation of liver enzymes and bilirubin was seen in the majority of patients. Only one patient was treated with 4.0 mg kg(-1) L-PAM, who died 8 days after IHP as a result of multiple-organ failure. A statistically significant correlation was found between the dose of L-PAM, peak L-PAM concentrations in perfusate (R = 0.86, P< or =0.001), perfusate area under the concentration-time curve (AUC; R = 0.82, P<0.001), tumour tissue concentrations of L-PAM (R = 0.83, P = 0.011) and patient survival (R = 0.52, P = 0.02). The peak L-PAM concentration and AUC of L-PAM in perfusate at dose level 3.0 mg kg( 1) (n = 5) were respectively 35- and 13-fold higher than in the systemic circulation, and respectively 30- and 5-fold higher than reported for high dose oral L-PAM (80-157 mg m(-2)) and autologous bone marrow transplantation. Median survival after IHP (n = 21) was 19 months and the overall response rate was 29% (17 assessable patients; one complete and four partial remissions). Thus, the maximally tolerated dose of L-PAM delivered via IHP is approximately 3.0 mg kg( 1), leading to high L-PAM concentrations at the target side. Because of the complexity of this treatment modality, IHP has at present no place in routine clinical practice. PMID- 10789722 TI - Persistent gestational trophoblastic disease: results of MEA (methotrexate, etoposide and dactinomycin) as first-line chemotherapy in high risk disease and EA (etoposide and dactinomycin) as second-line therapy for low risk disease. AB - Persistent gestational trophoblastic disease is potentially fatal, but the majority of patients are cured with chemotherapy. Any developments in treatment are therefore being directed towards maintaining efficacy and reducing toxicity. We evaluated efficacy and toxicity of methotrexate, etoposide and dactinomycin (MEA) as first-line therapy for high risk disease and etoposide and dactinomycin (EA) as second-line therapy for methotrexate-refractory low risk disease in a retrospective analysis of 73 patients (38 MEA, 35 EA) treated since 1986 at a supra-regional centre. The median follow-up period was 5.5 years and the median number of cycles received was 7. The overall complete response rate was 85% (97% for EA, 75% for MEA). Of eight patients who failed to respond, four have since died and four were cured with platinum-based chemotherapy. Alopecia was universal. Grade II or worse nausea, emesis, or stomatitis was observed in 29%, 30% and 37% respectively. Fifty-one per cent experienced grade II/III anaemia, 8% grade II or higher thrombocytopenia and 64% grade III or IV neutropenia; in six cases this was complicated by sepsis. Fifty-four per cent of patients went on to have a normal pregnancy. No patient has developed a second malignancy. In conclusion, the MEA and EA chemotherapy regimens for persistent trophoblastic disease are very well tolerated, do not appear to affect future fertility and are associated with excellent, sustained complete response rates. PMID- 10789723 TI - The increase in bladder carcinoma cell population induced by the free beta subunit of human chorionic gonadotrophin is a result of an anti-apoptosis effect and not cell proliferation. AB - Ectopic production of free beta human chorionic gonadotrophin (hCGbeta) by bladder carcinoma is well described and occurs in approximately 35% of cases. hCGbeta secreting tumours are more aggressive, radioresistant and have a greater propensity to metastasize. We proposed that the ectopic production of hCGbeta was contributing in an autocrine fashion to the radioresistance and metastatic potential of such secreting tumours. Though we demonstrated that the addition of hCGbeta to the culture media of bladder, cervical and endometrial carcinoma cell lines brought about an increase in cell populations this was not accompanied by a significant increase in the rate of replication. Since a cell population size is a balance of mitosis and mortality, we proposed that hCGbeta was inhibiting apoptosis. Here we have demonstrated that following incubation with recombinant hCGbeta, bladder carcinoma cells refrain from undergoing apoptosis. Quantitation of apoptotic bodies was carried out by immunoassay and corrected to cell number as determined by MTT assay. In each cell line, addition of hCGbeta reduced the number of apoptotic bodies dose-dependently, indicating a diminished apoptotic rate. Furthermore, TGFbeta1-induced apoptosis could be dose-dependently inhibited by co-incubation with hCGbeta. We propose, therefore, that such a decline in apoptosis may account for the cell population increase previously reported. It may also explain the radioresistance and aggressive nature of hCGbeta-secreting tumours and the poor prognosis associated therein. PMID- 10789724 TI - A dominant negative mutation of transforming growth factor-beta receptor type II gene in microsatellite stable oesophageal carcinoma. AB - Recent investigations revealed microsatellite instability in colon cancers are associated with mutations of the transforming growth factor-beta receptor type II gene (TGF-beta RII) that encodes a transmembrane protein containing an intracellular serine/threonine kinase domain. Activation of TGF-beta receptor type I (RI) and RII by TGF-beta induces nuclear translocation of Smad proteins including Smad2 and Smad4 that have been originally identified as tumour suppressor genes. We have previously reported six cases with microsatellite instability in 32 oesophageal carcinomas. In this study, we analysed genetic mutations of TGF-beta RII, Smad2 and Smad4 in these oesophageal carcinoma tissues and established 16 cell lines. No genetic mutation was detected in any tissues or cell lines except one tissue sample of microsatellite stable oesophageal carcinoma, that is, a mis-sense mutation of glutamic acid to glutamine at codon 526 (E526Q) in the TGF-beta RII serine/threonine kinase domain. Interestingly, the mutant TGF-beta RII E526Q can completely inhibit TGF-beta-induction of nuclear translocation of Smad4 protein in oesophageal carcinoma cells. This mutation of TGF-beta RII that is not associated with microsatellite instability might make a dominant negative effect on TGF-beta signal transduction in oesophageal carcinoma. PMID- 10789726 TI - Factors affecting survival in White and Asian children with acute lymphoblastic leukaemia. AB - Some studies suggest that Asian children with leukaemia have a worse outcome than Whites. Survival of Asians with ALL treated at the Birmingham Children's Hospital from 1975 to 1994 was the same as that of Whites, despite their greater deprivation and poorer nutrition. For one 5-year period (1980-1984) Asians had significantly poorer survival, even after adjustment for prognostic factors. Poor treatment compliance during that period may have contributed to this difference. PMID- 10789725 TI - Altered expression of members of the IGF-axis in hepatoblastomas. AB - Previous reports have demonstrated that expression of insulin-like growth factor 2 (IGF2) is altered in hepatoblastoma. Using RNAase protection analysis (RPA), we examined the gene expression for IGF1, IGF2, IGF1R, M6P/IGF2R, IGFBP-1 and IGFBP 2 in a series of hepatoblastomas with corresponding normal liver from the same individuals. The results show that the expression of the IGF-axis members included in the present study are altered between tumour and normal, and indicate that the IGF-axis may be involved in hepatoblastoma development. PMID- 10789727 TI - Space-time clustering patterns in childhood leukaemia support a role for infection. AB - Previous studies of space-time clustering in childhood leukaemia have produced equivocal and inconsistent results. To address this issue we have used Manchester Children's Tumour Registry leukaemia data in space-time clustering analyses. Knox tests for space-time interactions between cases were applied with fixed thresholds of close in space, <5 km and close in time <1 year apart. Addresses at birth as well as diagnosis were utilized. Tests were repeated replacing geographical distance with distance to the Nth nearest neighbour. N was chosen such that the mean distance was 5 km. Data were also examined by a second order procedure based on K-functions. All methods showed highly significant evidence of space-time clustering based on place of birth and time of diagnosis, particularly for all leukaemias aged 0-14 and 0-4 years, and acute lymphoblastic leukaemia (ALL) 0-4 years. Some results based on location at diagnosis were significant but mainly gave larger P-values. The results are consistent with an infectious hypothesis. Furthermore, we found an excess of male cases over females involved in space-time pairs. We suggest this may be related to genetic differences in susceptibility to infection between males and females. These findings provide the basis for future studies to identify possible infectious agents. PMID- 10789728 TI - Urinary androgens and breast cancer risk: results from a long-term prospective study based in Guernsey. AB - Between 1961 and 1967 a cohort of over 5000 women volunteered for a prospective study to determine the relationship between the urinary androgen metabolites, androsterone (A) and aetiocholanolone (E), and risk of breast cancer. During the first 10 years of the study the concentration of urinary A and E was determined in 1887 of the urine specimens. In 1971 we reported that subnormal amounts of urinary A and E were associated with a significantly increased risk of breast cancer. The cohort has been followed regularly during the 37 years since inception of the study and, by May 1998, 248 women had been diagnosed with breast cancer. Urinary androgen metabolites had been measured in 116 of these cases. Analysis of these data confirmed that women diagnosed in the first decade of the study were more likely to have low levels of urinary androgen metabolites. In the following decades, however, those who developed breast cancer were more likely to have manifested an increased A and E excretion. The reversal in the relationship between androgen metabolite excretion and risk suggests that age, or probably more importantly, menopausal status at diagnosis is an important modifying factor. Dichotomizing at age 50 it was found that in the younger age group (predominantly premenopausal) the rate ratios in the lowest tertile of A or E excretion were two- to threefold greater than for those in the highest tertile (chi2(1) = 3.57; P = 0.06: chi2(1) = 4.70; P = 0.03 for A and E respectively). In contrast, in the older age group comprising predominantly post-menopausal women, the rate ratios associated with the lowest tertile of A or E were half that of those in the highest tertile (chi2(1) = 4.10; P = 0.04; chi2(1) = 8.72; P = 0.003 for A and E respectively). This suggests that there may be different endocrine promotional factors for pre-and post-menopausal breast cancer. Hormonal risk factors may vary during the lifetime of an individual woman and this may have profound consequences for prevention strategies. PMID- 10789729 TI - Trends in cancer incidence in Kyadondo County, Uganda, 1960-1997. AB - Incidence rates of different cancers have been calculated for the population of Kyadondo County (Kampala, Uganda) for four time periods (1960-1966; 1967-1971; 1991-1994; 1995-1997), spanning 38 years in total. The period coincides with marked social and lifestyle changes and with the emergence of the AIDS epidemic. Most cancers have increased in incidence over time, the only exceptions being cancers of the bladder and penis. Apart from these, the most common cancers in the early years were cervix, oesophagus and liver; all three have remained common, with the first two showing quite marked increases in incidence, as have cancers of the breast and prostate. These changes have been overshadowed by the dramatic effects of the AIDS epidemic, with Kaposi's sarcoma emerging as the most common cancer in both sexes in the 1990s, and a large increase in incidence of squamous cell cancers of the conjunctiva. In the most recent period, there also seems to have been an increase in the incidence of non-Hodgkin lymphomas. So far, lung cancer remains rare. Cancer control in Uganda, as elsewhere in sub-Saharan Africa, faces a threefold challenge. With little improvement in the incidence of cancers associated with infection and poverty (liver, cervix, oesophagus), it must face the burden of AIDS-associated cancers, while coping with the emergence of cancers associated with Westernization of lifestyles (large bowel, breast and prostate). PMID- 10789730 TI - Risk of cutaneous malignant melanoma in relation to use of sunbeds: further evidence for UV-A carcinogenicity. AB - In a population-based, matched, case-control study from southern Sweden of 571 patients with a first diagnosis of cutaneous malignant melanoma and 913 healthy controls aged 16-80 years, the association between sunbed use and malignant melanoma was evaluated. A total of 250 (44%) cases and 372 (41%) controls reported ever having used sunbeds. A significantly elevated odds ratio for developing malignant melanoma after regular exposure to sunbeds was found, adjusted for hair colour, raised naevi, skin type and number of sunburns (odds ratio (OR) 1.8, 95% confidence interval (CI) 1.2-2.7). A dose-response relationship between total number of sunbed uses and melanoma risk was only found up to the level of 250 times. The OR was higher in individuals younger than age 36 years (adjusted OR 8.1, 95% CI 1.3-49.5 for regular vs. never use). The association seemed to be true only for subjects with black/dark brown or light brown hair and among females. Lesions of the extremities showed the strongest association of increased risk with sunbed use. An increased risk was related to commercial exposure and to exposure during the winter. The results substantiate the hypothesis that exposure to sunbeds might increase the risk of developing malignant melanoma. PMID- 10789731 TI - Tubal ligation and risk of breast cancer. AB - Although it has been demonstrated in previous studies that tubal ligation can have widespread effects on ovarian function, including a decrease in the risk of subsequent ovarian cancer, few studies have evaluated effects on breast cancer risk. In a population-based case-control study of breast cancer among women 20-54 years of age conducted in three geographic areas, previous tubal ligations were reported by 25.3% of the 2173 cases and 25.8% of the 1990 controls. Initially it appeared that tubal ligations might impart a slight reduction in risk, particularly among women undergoing the procedure at young ages (<25 years). However, women were more likely to have had the procedure if they were black, less educated, young when they bore their first child, or multiparous. After accounting for these factors, tubal ligations were unrelated to breast cancer risk (relative risk (RR) = 1.09, 95% confidence interval (CI) 0.9-1.3), with no variation in risk by age at, interval since, or calendar year of the procedure. The relationship of tubal ligations to risk did not vary according to the presence of a number of other risk factors, including menopausal status or screening history. Furthermore, effects of tubal ligation were similar for all stages at breast cancer diagnosis. Further studies would be worthwhile given the biologic plausibility of an association. However, future investigations should include information on type of procedure performed (since this may relate to biologic effects) as well as other breast cancer risk factors. PMID- 10789732 TI - Feasibility of targeted early detection for melanoma: a population-based screening study. AB - The feasibility of targeted screening for cutaneous malignant melanoma in the UK using a postal questionnaire and invitation to screening by a consultant dermatologist was investigated in a population based cross-sectional survey. A total of 1600 people aged 25-69 years, stratified by the social deprivation score of wards within one general practice, were randomly selected from a population of 8000.1227 (77%) returned the questionnaire and 896 (56%) attended the screening clinic. Uptake was lower for men (P<0.001), those aged under 50 (P<0.001), people from deprived areas (P<0.001) and skin types III and IV (men only, P<0.001). Twenty per cent of women and 10% of men felt nervous about attending the clinic, but only 4% were worried by the questionnaire. The level of agreement between the self- and dermatologist's assessments of risk factors was best for hair colour (Kappa = 0.67, sensitivity 73% and specificity 98%). People tended to under report their level of risk. Over 95% knew about at least one major sign, but 54% reported incorrect signs of melanoma. Targeted screening for melanoma in the UK will be hampered by difficulties in accurately identifying the target population. Strategies to improve skin self-awareness rather than screening should be developed and evaluated. PMID- 10789733 TI - Induction of T-cell mitogenic unresponsiveness by recombinant human granulocyte colony-stimulating factor (rHuG-CSF) PMID- 10789734 TI - Methods in molecular biology: minor errors in primer citations with major consequences: how can we minimize these mistakes? PMID- 10789735 TI - Prognostic value of histological and biological markers in pharyngeal squamous cell carcinoma: a case-control study. PMID- 10789736 TI - Beatson International Cancer Conference: invasion and metastasis. PMID- 10789737 TI - Prior pregnancy ameliorates the course of intra-abdominal desmoid tumors in patients with familial adenomatous polyposis. AB - PURPOSE: Intra-abdominal desmoid tumors occur in 12 percent of patients with familial adenomatous polyposis. A minority grow quickly and are lethal, most are relatively inert, and some cause problems by obstructing adjacent organs. Desmoid tumors may be estrogen-dependent, and estrogen-blocking drugs are part of the usual treatment of these tumors. This study was performed to examine the effect of pregnancy on the course of patients with familial adenomatous polyposis and intra-abdominal desmoids. METHODS: All females with familial adenomatous polyposis and an intra-abdominal desmoid treated or followed up at this institution were eligible. Stable, asymptomatic desmoids were followed up yearly with examination and CT scan. Growing or symptomatic desmoids were followed up at least every six months. Maximum tumor size was grouped as follows: <10 cm, 10 to 20 cm, and >20 cm. A change in tumor size was defined as a change of +/-50 percent or more of maximum diameter. Stable tumors showed no change in diameter during the study period; variable growth was defined as a significant change in either direction that was followed by a return to previous dimensions or a stabilization of growth. Rapid growth was a doubling of diameter within three months. Pregnant females were compared with nonpregnant females. Subgroups of females were matched for age at diagnosis of desmoid. RESULTS: Twenty-two females had never been pregnant, whereas 25 had been pregnant at least once. Eleven pairs were matched for age. There were no differences between groups in the incidence of extracolonic manifestations of familial adenomatous polyposis, family history of desmoids, number or type of surgeries done for familial adenomatous polyposis, length of follow-up, or time from surgery to desmoid diagnosis. Desmoids in pregnant females had a significantly more benign course: 18 were stable (vs. 6 nonpregnant females), 2 had variable growth (vs. 10), 1 had rapid growth (vs. 5), and 4 disappeared (vs. 1). There were also trends to smaller, less symptomatic tumors requiring treatment less often in pregnant females. CONCLUSIONS: Pregnancy seems to ameliorate the course of abdominal desmoid tumors significantly in females with familial adenomatous polyposis. This finding raises questions about the most appropriate hormonal treatment for these tumors. Perhaps progesterone or prolactin therapy should be tried, alone or in combination with estrogen. If further studies confirm these findings, females with a family history of desmoid tumors should not be advised against pregnancy. PMID- 10789738 TI - Molecular prognostic factors in rectal cancer treated by radiation and surgery. AB - PURPOSE: The aim of this study was to determine the biologic significance of tumor response and the prognostic value of molecular markers in a group of patients with rectal cancer treated with preoperative radiation therapy and radical surgery. METHODS: Microsatellite instability, microvessel count, and immunohistochemistry for proliferating cell nuclear antigen, p53, p21, bcl-2, and vascular endothelial growth factor were performed in the preradiation biopsy specimen of 72 patients with rectal cancer treated by preoperative radiation therapy and radical curative surgery. Preoperative tumor stage by endorectal ultrasound was compared with pathology stage of the resected specimen. Mean follow-up was 50 months. RESULTS: Twenty-eight patients (39 percent) responded to radiation therapy. The response was complete in 8 (12 percent) and partial in 20 patients (27 percent). Tumors with positive nodes in the surgical specimen were less likely to have responded to preoperative radiation (P = 0.03). Only p21 expression was individually associated with response to radiation (56 vs. 30 percent; P = 0.03). Tumors that were p53-negative/p21-positive or p21 positive/bcl-2-positive were also more likely to respond to radiation (83 vs. 35 percent; P = 0.03 and 71 vs. 31 percent; P = 0.01, respectively). The tumor relapsed in 21 patients (29 percent): locally in 7 (10 percent) and distally in 14 (19 percent). Recurrence was associated with lack of response to radiation, female gender, distal tumor location, high proliferating cell nuclear antigen labeling index, and low microvessel count. Probability of survival was greater for patients with well or moderately differentiated tumors and tumors that responded completely to radiation. CONCLUSIONS: Tumor response to radiation is associated with improved tumor control and overall survival rate, and p21 expression is a marker of tumor radiosensitivity in patients with rectal cancer. Furthermore, a high proliferating cell nuclear antigen labeling index and a low microvessel count in the preradiation biopsy specimen may be prognostic indicators for tumor recurrence. PMID- 10789739 TI - Pancreatic or duodenal resection or both for advanced carcinoma of the right colon: is it justified? AB - PURPOSE: The role of extended resections for locally advanced carcinomas of the right colon infiltrating duodenum and pancreas remains unclear. This investigation was undertaken to review our experience with pancreatic head or duodenal resections for advanced right-sided colon cancer. METHODS: The clinical, pathologic, and follow-up details of eight patients with bulky primary carcinomas of the right colon infiltrating the duodenum (n = 4) or pancreatic head (n = 4) surgically managed at Memorial Sloan-Kettering Cancer Center between 1986 and 1998 were reviewed. RESULTS: Six patients presented with anemia, and one patient each with epigastric pain and an abdominal mass. All patients had T4 lesions, whereas five had lymph node metastases at presentation. All patients were resected with clear pathologic margins either by right colectomy and en bloc duodenectomy (n = 4), or en bloc pancreaticoduodenectomy (n = 4). The 30-day mortality rate was zero. Six patients remained alive and free of disease at a median follow-up of 26 months, and there was one long-term survivor who was alive and free of disease at 84 months after resection. CONCLUSION: Extended resection for localized primary colonic carcinoma invading pancreas or duodenum can be undertaken safely and is associated with prolonged survival time. PMID- 10789740 TI - Changing management trends in penetrating colon trauma. AB - PURPOSE: Recent prospective studies have recommended primary repair for all penetrating colon injuries. We evaluated our management trends given these recommendations and assessed our results of primary repair. METHODS: A retrospective review was conducted of 145 patients with penetrating colon injuries received between January 1, 1991, and December 31, 1997. The patients were characterized according to demographics and severity of injury. Morbidity was defined as failure of a primary repair, abscess, fistula, wound dehiscence, fasciitis, sepsis, organ failure, or coagulopathy. The periods 1991 to 1993 (early period) and 1994 to 1997 (late period) were chosen for comparison. RESULTS: Primary repairs were performed in 53 of 75 patients (71 percent) during the early period and in 61 of 70 patients (87 percent) during the late period (P = 0.03). No significant differences in demographics or injury severity were found to account for the increased rate of primary repairs. The number of suture repairs was nearly equal in both periods (59 vs. 61 percent). The number of resections and anastomoses for destructive colon injuries was significantly higher in the late period (26 percent) compared with the early period (12 percent; P = 0.05). Morbidity was equal (24 percent) in the two periods. There were no failures of resections and anastomoses and one failure of suture repair. CONCLUSIONS: Increased primary repair occurred because of more liberal use of resection and anastomosis for destructive injuries. Suture repair was performed for the amenable colonic injury throughout the study period. Risk factors for failure of resection and anastomosis cannot be defined from our study. Further investigation is needed to determine if resection and anastomosis is safe for the most severely injured patients. PMID- 10789741 TI - Postdelivery anal function in primiparous females: ultrasound and manometric study. AB - PURPOSE: A study was performed to evaluate the early morphologic and functional consequences of vaginal delivery on the anal sphincter in primiparous females. METHODS: Among a cohort of 197 primiparous females who agreed to participate in a clinical evaluation of fecal incontinence and in a transanal ultrasound examination 12 weeks after delivery, 52 also underwent anal manometry using a radial six-port catheter, of whom 10 were asymptomatic and had a normal sphincter at ultrasound and the remaining 42 had clinical signs of anal incontinence or ultrasonographic defects of the anal sphincter or both. Anal sphincter pressures and asymmetry index were analyzed at rest and during voluntary squeeze. Manometric and ultrasound results were compared, together with clinical symptoms. RESULTS: Fourteen patients with clinical signs of anal incontinence had lower resting and squeeze anal pressures than continent patients (P < 0.05), but similar anal asymmetry indexes. Patients with incontinence and an anal defect had the lowest resting and squeeze anal pressures (P < 0.05). Forceps assistance to delivery was not associated with a higher frequency of anal sphincter lesions. Resting and squeeze anal pressures were lower in the forceps group (P < 0.005), but anal asymmetry indexes were similar. Finally, manometric results were identical in the presence or absence of anal sphincter endosonographic defects. CONCLUSIONS: Anal sphincter defects are frequent after the first vaginal delivery, but are not always associated with functional or clinical abnormalities. Resting and squeeze anal pressures were significantly decreased in patients with incontinence and an anal defect and after forceps-assisted deliveries. Anal asymmetry index was not found useful in this population of young primiparous females. PMID- 10789742 TI - Prognostic significance of rectocele, intussusception, and abnormal perineal descent in biofeedback treatment for constipated patients with paradoxical puborectalis contraction. AB - PURPOSE: The findings of paradoxical puborectalis contraction, rectocele, sigmoidocele, intussusception, and abnormal perineal descent often coexist in constipated patients, as noted by defecographic study. Moreover, some of these conditions are often found in asymptomatic patients. Biofeedback is the treatment of choice for constipated patients with paradoxical puborectalis contraction; the main determinant of successful biofeedback is patient compliance. The significance of coexistent and highly prevalent variants, such as rectocele, intussusception, sigmoidocele, or abnormal perineal descent, on the success of biofeedback is unknown. This review was designed to assess whether these coexisting defecographic findings have any prognostic significance for the outcome of biofeedback. METHODS: From July 1988 to December 1996, 209 constipated patients with paradoxical puborectalis contraction underwent biofeedback treatment after defecography. A total of 173 patients (120 females) who had more than one biofeedback session after defecography formed the study group. Defecographic findings included concomitant rectoceles, 40 (23 percent); evidence of circumferential intussusception, 17 (10 percent); sigmoidocele, 13 (8 percent); and abnormal perineal descent, 109 (63 percent). RESULTS: Whereas 65 patients failed to complete the course of biofeedback therapy, 108 (62.4 percent) patients completed the course of biofeedback and were discharged by the therapist. Within the completed group 59 (55 percent) improved, and 49 (45 percent) patients failed biofeedback therapy. In the improved group 14 (23.7 percent) had a rectocele, 5 (8.5 percent) had intussusception, 5 (8.5 percent) had a sigmoidocele, and 37 (62.7 percent) had abnormal perineal descent. In the failure group 9 (18.4 percent) had a rectocele, 5 (10.2 percent) had an intussusception, 2 (4.1 percent) had a sigmoidocele, and 31 (63.3 percent) had abnormal perineal descent (P = not significant). The success of biofeedback was then analyzed relative to the number of coexisting conditions. Specifically, the outcome in patients with paradoxical puborectalis contraction alone and with one, two, and three other defecographic findings were compared. No statistically significant difference was found among these four groups. CONCLUSION: Although other defecographic findings frequently coexist with paradoxical puborectalis contraction, none of the concomitant findings adversely affected the outcome of biofeedback treatment. Therefore, biofeedback can be recommended to patients with coexistent defecographic findings, with expectation of success in over 50 percent of individuals who complete the course of therapy. PMID- 10789743 TI - Effectiveness of endoluminal sonography in the identification of occult local rectal cancer recurrences. AB - PURPOSE: Local recurrence of rectal cancer after curative surgery appears in 5 to 30 percent of all cases. It is necessary to detect local recurrence in a resectable stage to have an opportunity for curative reintervention or palliative prevention of those symptoms. Because most local recurrences occur extraluminally, conventional follow-up fails to detect them at an early stage. Therefore, a prospective study was performed to assess the diagnostic potential of endorectal and endovaginal ultrasound to detect asymptomatic resectable local recurrence. METHODS: In 338 patients 721 endoluminal ultrasound examinations were added to routine follow-up of rectal and left colonic cancer, with a mean of 2.1 (range, 1-10) endoluminal ultrasound examinations for each patient. RESULTS: A total of 116 patients (34.3 percent) were shown to have local recurrence, which was suggested by endoluminal ultrasound and proven by endoluminal ultrasound guided needle biopsy in all cases of unclear pararectal structures that could not be verified by endoscopic biopsy. Digital examination failed to detect local recurrence in 91 patients, endoscopy failed to detect local recurrence in 80 patients, and the levels of tumor markers were normal in 25 patients with confirmed local recurrence. In 33 cases of local recurrence, both digital examination and endoscopy results were normal. Twenty-five patients, in whom carcinoembryonic antigen levels, digital examination, and endoscopy results were normal, underwent potential curative reoperation, with total resection of the local recurrence. All 25 patients were still alive at the end of the study period, and 21 were free from disease. On the other hand, only 6 of 67 patients with local recurrence detectable by conventional follow-up could be operated on with curative intention. CONCLUSION: Postoperative endoluminal ultrasound is able to detect local recurrence at an earlier and asymptomatic stage and can be verified by endosonography-guided needle biopsy. Routine use in follow-up may raise the ratio of curative retreatment by early detection of extramural local recurrence. PMID- 10789744 TI - Effect of surgeon specialty interest on patient outcome after potentially curative colorectal cancer surgery. AB - OBJECTIVE: There are significant differences in patient outcome after potentially curative surgery for colorectal cancer that relate to the surgeon performing the procedure. The reasons for these differences remain obscure. The aim of this study was to examine the effect of the surgeon's specialty on patient outcome after potentially curative colorectal cancer surgery and to identify factors that may help explain differences in outcome among specialty groups. METHODS: Between 1990 and 1993, 378 patients underwent potentially curative surgery for colorectal cancer by surgeons with different specialty interests, vascular or transplant, general, and colorectal surgeons, in a large teaching hospital. Information on operative details, including the length of the resection specimen, resection margins, whether the tumor was removed with en bloc resection of adjacent clinically involved organs, number of lymph nodes removed, and stage was collected. Factors affecting both local and overall recurrence rates were analyzed using logistic regression analysis at both univariate and multivariate levels. RESULTS: At a median follow-up of 45 months the only factors associated with a significantly reduced local recurrence rate were the length of the resection specimen (odds ratio, 0.56; 95 percent confidence interval, 0.31-0.99) and colorectal specialty (P = 0.04). Patients operated on by a general surgeon were 3.42 times (95 percent confidence interval, 1.32-8.9) more likely to develop a local recurrence than those operated on by a colorectal surgeon. For overall recurrence, early stage disease (P < 0.0001), absence of vascular invasion (0.005), and colorectal specialty (0.025) were the only factors associated with significantly improved outcome at multivariate analysis. CONCLUSIONS: These data show that surgeons with an interest in colorectal cancer achieve lower local and overall recurrence rates compared with vascular or transplant or general surgeons. Differences in local recurrence rates seem to be predominantly related to the extent of resection performed and demonstrate the need to remove an adequate specimen when performing potentially curative colorectal cancer surgery. PMID- 10789745 TI - Photodynamic therapy for residual neoplasms of the perianal skin. AB - PURPOSE: The aim of this study was to evaluate the efficacy of photodynamic therapy in the management of residual neoplasms of the perianal skin. METHODS: This is a retrospective review. Five patients with pathologic confirmation of residual perianal neoplasms were treated with photodynamic therapy. There were three females. The mean age was 52 (range, 33-79) years. Pathology consisted of Bowen's disease in two patients, squamous-cell carcinoma in two patients, and extramammary Paget's disease in one patient. Four patients received one photodynamic therapy treatment and one patient received two treatments three months apart. RESULTS: Treatment was followed by immediate perianal erythema, subsequent blister formation in 36 to 48 hours, and sloughing of the treated area in 72 hours. With a mean follow-up of 5.2 (range, 1-8) years, there were two recurrences. One recurrence was in a patient four years after treatment for Paget's disease, and the other was in a patient nine months after treatment for Bowen's disease. The latter was managed successfully with wide local excision. Treatment-related toxicities included significant perianal pain in four patients, controlled with analgesia management. CONCLUSIONS: Photodynamic therapy can successfully be used after wide local excision for residual neoplasms of the perianal skin. Treatment can be rendered with acceptable morbidity. PMID- 10789746 TI - Hemorrhoidectomy vs. Lord's method: 17-year follow-up of a prospective, randomized trial. AB - PURPOSE: A prospective, randomized trial was performed between 1979 and 1981 comparing anal dilation and hemorrhoidectomy for hemorrhoidal disease at the Maastricht University Hospital. The aim of this study was to update that trial to assess long-term outcome and complications such as fecal incontinence. METHODS: A total of 138 patients with second-degree and third-degree hemorrhoids entered the study. Median follow-up was 17 (range, 8.4-18.3) years and was achieved for 118 (86 percent) patients. Group A (n = 35) underwent hemorrhoidectomy and Group B (n = 39) was treated with anal dilation and aftertreatment (original Lord's procedure), whereas Group C (n = 44) had dilation only. Fourteen of these patients died. RESULTS: Recurrent hemorrhoids were noted for 26 percent of the patients treated with hemorrhoidectomy, for 46 percent with operative dilation with the postoperative dilation program, and for 39 percent with operative dilation without the postoperative program. The percentage of repeated treatment for the three subgroups was 11, 23, and 18 percent, respectively. The continence status remained more or less the same during the first year. However, 17 years later the anal stretch procedures caused various incontinence disorders in 52 percent of these patients. Significance was found for incontinence of flatus (from 11 to 30 patients; P = 0.04) in the anal dilation groups. CONCLUSIONS: Hemorrhoidectomy can be considered to be a safe procedure for treatment of hemorrhoidal disease, with excellent long-term results. Anal dilation is associated with a high percentage of complaints of fecal incontinence. The procedure should be abandoned. PMID- 10789747 TI - Premedication with dextromethorphan provides posthemorrhoidectomy pain relief. AB - PURPOSE: Previous studies have shown that N-methyl-D-aspartate receptor antagonists provide a preemptive analgesic effect in humans. This study was designed to examine whether premedication with dextromethorphan, an N-methyl-D aspartate antagonist, also provided a preemptive analgesic effect that improved postoperative pain management. METHODS: Sixty patients who were American Society of Anesthesiologists status I and II scheduled for hemorrhoidectomy (modified Whitehead procedure) were included in the study. Patients were randomly assigned to the control and study groups. For the control group patients received chlorpheniramine maleate (20 mg), a component of the injection form of dextromethorphan, intramuscular injection 30 minutes before skin incision. In the study group dextromethorphan 40 mg containing 20 mg chlorpheniramine maleate (intramuscular) was given as premedication 30 minutes before skin incision. Pethidine (1 mg/kg, intramuscular) was given for pain relief as required postoperatively. The time to first pethidine injection, total pethidine consumption, worst pain score, and pethidine-related side effects were recorded for 48 hours postoperatively. RESULTS: The times to first pethidine injection (mean +/- standard error of the mean) were 5.2 +/- 3 and 19.6 +/- 6 hours in the control and study groups, respectively. Total pethidine consumption was 140 +/- 11.3 and 63.5 +/- 11.8 mg in the control and study groups. The worst visual analog scale pain scores were 7.4 +/- 0.2 and 5.6 +/- 0.3 in the control and study groups during the two-day observation. The numbers of patients who required pethidine injection were 29 and 20 in the control and study groups, respectively. Two patients suffered pethidine-related side effects, such as nausea, vomiting, dizziness, and headache, in the control group, and no patient complained of any side effect in the study group. CONCLUSION: We found that dextromethorphan premedication provided a preemptive analgesic effect, thus producing reduced postoperative pain and pethidine requirement and improved recovery from hemorrhoidectomy. PMID- 10789748 TI - Prognostic value of magnetic resonance imaging in the management of fistula-in ano. AB - PURPOSE: Magnetic resonance imaging of fistula-in-ano has been shown to predict surgical anatomy accurately and identify complex features. In addition, fistula complexity has been correlated with poor outcome after surgical intervention. We investigated whether preoperative magnetic resonance imaging could predict clinical outcome after surgery for fistulous disease better than clinical examination under anesthetic. METHODS: Seventy patients with clinically suspected fistula-in-ano underwent preoperative dynamic contrast-enhanced magnetic resonance imaging before surgical exploration. Outcome was assessed at a minimum of one year after surgical exploration and correlated in a blinded fashion with the surgical and magnetic resonance grading of the severity of the fistulous disease. RESULTS: Of 70 patients, 12 were not operated on and 6 were lost to follow-up, making 52 patients eligible for analysis. Assessment by dynamic contrast-enhanced magnetic resonance imaging more accurately predicted outcome than the findings at initial surgical exploration. Dynamic contrast-enhanced magnetic resonance imaging had a sensitivity of 81 percent, specificity of 73 percent, and positive predictive value of 75 percent; surgery had a sensitivity of 77 percent, specificity of 46 percent, and positive predictive value of 59 percent. Surgical assessment of apparent disease severity bore no relation to final outcome. Dynamic contrast-enhanced magnetic resonance imaging could accurately predict whether patients were likely to have a satisfactory or unsatisfactory outcome after surgery. CONCLUSION: Dynamic contrast-enhanced magnetic resonance imaging better predicts clinical outcome of patients with fistula-in-ano than initial surgical exploration. PMID- 10789749 TI - Coping behavior and social support contribute independently to quality of life after surgery for inflammatory bowel disease. AB - PURPOSE: The purpose of this study was to examine the association between coping behavior at the time of surgery and inflammatory bowel disease-related quality of life after surgery. We also investigated the relationship between perceived social support and both coping style and postsurgical quality of life. Finally, the value of the Medical Outcomes Study Social Support Scale for preoperative screening was assessed. METHODS: Eighty-six subjects who had surgery during a 12 month period completed the Inflammatory Bowel Disease Questionnaire, the Ways of Coping, a measure of inflammatory bowel disease symptom severity, and the Medical Outcomes Study Social Support Scale. Analysis of variance was used to test an association between Ways of Coping score and membership in a high quality of life (Inflammatory Bowel Disease Questionnaire > mean) or low quality of life (Inflammatory Bowel Disease Questionnaire < mean) cohort. Comparison of group means between the high quality of life and low quality of life cohorts identified Ways of Coping behavior scales that differed between the high quality of life and low quality of life cohorts. Stepwise linear regression analysis was then used to determine the independent contribution of 1) current inflammatory bowel disease symptoms, 2) current perceived social support, and 3) identified coping behaviors (self-control, self-blame, and escape, summed as a single index named "maladaptive coping") to postsurgical quality of life. The sensitivity, specificity, and negative predictive value of the Medical Outcomes Study Social Support Scale were assessed. RESULTS: The lower quality of life group distinguished itself by more frequent use of maladaptive coping. Regression analysis revealed that current inflammatory bowel disease-related symptoms, current perceived social support, and maladaptive coping behaviors at the time of surgery each made a highly significant independent contribution to postsurgical quality of life. The sensitivity of the Medical Outcomes Study Social Support Scale in identifying patients with poor postsurgical quality of life was 81 percent, and the specificity was 77 percent. The negative predictive value was 93 percent. CONCLUSIONS: Three coping behaviors which seem to be maladaptive (self control, self-blame, and escape) are associated with lower quality of life after surgery for inflammatory bowel disease. These coping behaviors make a contribution to postsurgical quality of life independent of the negative effect on quality of life of inflammatory bowel disease symptoms. Perceived social support is a third factor that makes an independent contribution to postsurgical quality of life. The Medical Outcomes Study Social Support Scale has properties associated with an effective screening tool and merits further investigation as an instrument to screen presurgically for individuals at higher risk of poor subjective outcome of inflammatory bowel disease surgery. PMID- 10789750 TI - "Dog ear" formation after double-stapled low anterior resection as a risk factor for anastomotic disruption. AB - PURPOSE: The aim of this study was to investigate the possible deleterious effect of the lateral intersecting margins (so-called dog ears) on anastomotic disruption after experimentally performed double-stapled anastomoses. METHODS: Two groups of double-stapled side-to-end anastomoses were performed using pig small intestines. Group A consisted of 35 circular anastomoses and Group B of 32 double-stapled anastomoses with a bilateral dog ear. In both groups bursting pressures were tested using a water-filled, pressure-controlled automatic pumping system (Hamou Endomat), and special attention was paid to the location(s) in the anastomoses were the disruption(s) occurred. RESULTS: In Group A bursting pressures were significantly higher than in Group B (median pressure, 90 vs. 60 mmHg; P < 0.001, Mann-Whitney U test). Remarkably, in Group B in 13 cases (42 percent) the first disruption occurred at the corner of a dog ear. CONCLUSIONS: We conclude that the lateral intersections of double-stapled anastomoses are a structural weak spot and that the currently most often applied double-stapled anastomosis is a less effective type of anastomosis than a complete circular one. Resolving this technical problem might help to reduce the number of anastomotic disruptions after low anterior resections. PMID- 10789751 TI - Alteration of colonic mucin after ureterosigmoidostomy. AB - PURPOSE: Patients undergoing urinary diversion by ureterosigmoidostomy after complete cystectomy for malignant bladder tumors show a high incidence of neoplasia at and near the site of anastomosis. We examined a risk factor for tumor occurrence in the area of anastomosis, alterations of mucus glycoproteins in the surrounding colonic mucosa. METHODS: Colonoscopy was performed in 37 patients who had undergone ureterosigmoidostomy. Biopsy specimens were obtained near the ureteral anastomosis and were stained with hematoxylin and eosin, high iron-diamine alcian blue (pH 2.5), and a fluorescent lectin conjugate (peanut agglutinin). RESULTS: At the anastomotic site colonoscopy showed protruding lesions in 26 of 37 patients (71 percent), all histologically representing inflammatory granulomas. The mucosa around the anastomosis was normal in endoscopic appearance; however, histologically, slight inflammatory cell infiltration, edema, and increased numbers of Paneth cells were observed. Alcian blue staining revealed an increase in mucosal sialomucin postoperatively compared with preoperatively. The proportion of peanut agglutinin-binding mucin, not observed in normal mucosa but seen in malignant or premalignant tissue, was increased. CONCLUSION: As postoperative interval increases, changes in properties of the "background" mucosa become greater, which suggests an association with colonic carcinogenesis. PMID- 10789752 TI - Evaluation of transarterial embolization for lower gastrointestinal bleeding. AB - INTRODUCTION: Transcatheter arterial embolization has been used as a therapeutic maneuver for lower gastrointestinal bleeding. The availability of highly selective arteriography has made this procedure safer and warrants re-evaluation. METHODS: A retrospective chart review was done of all patients undergoing arteriography for presumed lower gastrointestinal bleeding at two acute-care community hospitals. Causes of bleeding, clinical outcome, and complications caused by transcatheter arterial embolization were recorded. RESULTS: There were 26 arteriographically identified bleeding sites in the colon and small bowel. The most frequent cause of bleeding was diverticulosis (12 patients), with the diagnosis being arterio venous malformation in two, and one unknown colonic source. Transcatheter arterial embolization was attempted for 17 separate bleeding episodes in 16 patients. Transfusion requirements were an average (+/- standard deviation) of 7 +/- 1.43 units per patient. Transcatheter arterial embolization was successful in stopping bleeding in 14 cases (82 percent). Two patients had surgery after transcatheter arterial embolization: one for colonic necrosis and one for persisting bleeding. There were two more unsuccessful procedures; one had a successful repeated transcatheter arterial embolization, and one stopped spontaneously. One patient rebled during the same hospitalization and was controlled with intra-arterial vasopressin. There were two deaths, both secondary to sepsis unrelated to the transcatheter arterial embolization or the gastrointestinal tract. CONCLUSIONS: Transcatheter arterial embolization is a relatively safe and successful procedure in patients with massive lower gastrointestinal hemorrhage. It is an excellent choice of therapy for patients that are poor candidates for surgery, but its role in other patients remains to be defined. PMID- 10789753 TI - Psychologic aspects in proctalgia. AB - PURPOSE: One of the main problems in coloproctology is chronic idiopathic anal pain. The aim of this study was to investigate the psychosomatic components of proctalgia to identify which, if any, component is associated with this pain and to what extent. METHODS: Twenty patients with proctalgia were observed (mean age, 46 years). Psychologic consultations were required by the surgeons, because of persistent symptoms, to allow a better understanding of the problem and a more integrated therapy. The psychologic investigation consisted of three interviews and administration of the following tests: Institute for Personality and Ability Testing Anxiety Scale Questionnaire (1-10), Rorschach test (Klopfer and Davidson method), and Draw-A-Person test by Karen Machover. This sample was compared with a control group composed of 40 healthy subjects, homogeneous in age, social and working conditions, and investigation procedures. RESULTS: Patients showed depression and anxiety according to standard validated questions (Institute for Personality and Ability Testing Anxiety Scale Questionnaire) and personality disorders; they had a strong tendency to use primitive defense mechanisms and showed a lack of personality formation. CONCLUSIONS: Psychologic investigation allows a progressive clarification of all the components of anal pain. This might be useful not only for research purposes but also for a more effective approach to these patients. PMID- 10789754 TI - Lymphoma arising in an S-pouch after total proctocolectomy for ulcerative colitis: report of a case. AB - The gastrointestinal tract is the most common extranodal site of primary non Hodgkin's lymphoma. We present a case of a 50-year-old male with primary B cell lymphoma arising in an S-pouch eight years after a total proctocolectomy for ulcerative colitis. After chemoradiotherapy the patient remained asymptomatic, with an intact S-pouch. Pouch conservation is feasible in patients with primary lymphoma of the pouch, using chemoradiotherapy and close follow-up examinations. PMID- 10789755 TI - Is Hartmann's procedure safe in Crohn's disease? Report of three cases. AB - INTRODUCTION: Crohn's disease-associated colorectal cancer may occur in an area of defunctioning bowel. Some patients with Crohn's colitis undergo subtotal colectomy, ileostomy, and low Hartmann's procedure in an effort to preserve the rectum. This procedure has also been advocated for patients with severe anorectal Crohn's disease, in whom nonhealing of the perineal wound after proctectomy occurs with alarming frequency. The authors present a review of the literature and three cases of cancer developing in the defunctioning rectal stump despite surveillance proctoscopy. METHODS: Twenty-five patients underwent low Hartmann's procedure for severe anorectal Crohn's disease. Surveillance proctoscopy was performed as follow-up. Development of cancer in the rectal remnant or anus or recurrence of symptoms was managed by resection and adjuvant therapy. RESULTS: One patient developed squamous-cell carcinoma of the anal canal, underwent resection and adjuvant therapy, and was disease free at the time of this study. Two patients developed adenocarcinoma of the rectum. Both underwent resection and adjuvant therapy. One patient died and the other developed a recurrence. CONCLUSIONS: The authors recommend interval perineal proctectomy in all patients undergoing low Hartmann's procedure for severe anorectal Crohn's disease in whom rectal preservation is not possible. Regularly scheduled interim surveillance proctoscopy performed every two years, with biopsies of macroscopically normal appearing and abnormal-appearing rectal mucosa and curetting of fistulous tracts, is also recommended to decrease the possibility of missing occult malignancies. PMID- 10789756 TI - Radical excision of multifocal anal intraepithelial neoplasia: report of a case. AB - PURPOSE: The aim of this study was to describe the technique of radical excision of multifocal anal intraepithelial neoplasia and discuss controversial issues surrounding the management of this condition. METHOD: We describe the case of a 31-year-old female with previous vulval warts, vulval squamous carcinoma, and recent immunosuppression who had widespread anal intraepithelial neoplasia excised, and the resulting defect was split-skin grafted, including the anal canal. RESULTS: Excision and split-skin grafting was successful in removing the disease and left a satisfactory cosmetic and functional result. CONCLUSION: Diffuse, high-grade, anal intraepithelial neoplasia is rare. Excision of these lesions remains controversial but may be the best option. PMID- 10789757 TI - Pseudomembranous enteritis after proctocolectomy: report of a case. AB - Intestinal pseudomembrane formation, sometimes a manifestation of antibiotic associated diarrheal illnesses, is typically limited to the colon but rarely may affect the small bowel. A 56-year-old female taking antibiotics, who had undergone proctocolectomy for idiopathic inflammatory bowel disease, presented with septic shock and hypotension. A partial small-bowel resection revealed extensive mucosal pseudomembranes, which were cultured positive for Clostridium difficile. Intestinal drainage contents from an ileostomy were enzyme immunoassay positive for C. difficile toxin A. Gross and histopathologic features of the small-bowel resection specimen were similar to those characteristic of pseudomembranous colitis. The patient was treated successfully with metronidazole. These findings suggest a reservoir for C. difficile also exists in the small intestine and that conditions for enhanced mucosal susceptibility to C. difficile overgrowth may occur in the small-bowel environment of antibiotic treated patients after colectomy. Pseudomembranous enteritis should be a consideration in those patients who present with purulent ostomy drainage, abdominal pain, fever, leukocytosis, or symptoms of septic shock. PMID- 10789758 TI - Adenocarcinoma arising in interposed colon: report of a case. AB - A rare occurrence of primary adenocarcinoma arising in an interposed colon nearly seven years after the surgery is described. Remarkably, there were no symptoms from a large mass in the transposed bowel. Diagnosis was made fortuitously while searching for the cause of the patient's iron-deficiency anemia. The rationale for careful preoperative screening of the colonic segment intended for transplant to exclude pre-existing pathology is reemphasized. The various methods of evaluating the postoperative colon graft and their advantages and limitations are discussed. PMID- 10789759 TI - Use of ultrasonic surgical aspirator in operative cytoreduction of pseudomyxoma peritonei. AB - Pseudomyxoma peritonei presents a unique challenge to the surgical oncologist. Residual gelatinous tumor with varying degrees of adherence always remains on the abdominal viscera after standard excisional therapy. Traditionally, this has been removed by "electroevaporation" with ball-tip diathermy, but this is associated with an extensive peritoneal burn and associated ileus. We describe the use of an ultrasonic surgical aspirator as a safe and efficient method of tumor removal in this condition. PMID- 10789760 TI - Routine anal cleansing, so-called hemorrhoids, and perianal dermatitis: cause and effect? PMID- 10789761 TI - Centennial dissertation. Percutaneous imaging-guided core breast biopsy: state of the art at the millennium. PMID- 10789762 TI - Centennial sounding board. Sonography: unlikely success, uncertain future. PMID- 10789763 TI - Diagnostic radiologists' subspecialization and fields of practice. AB - OBJECTIVE: The purpose of this study was to explore the diagnostic radiology profession from the perspective of subspecialization and fields of practice. MATERIALS AND METHODS: Data for 1244 practicing diagnostic radiologists were gathered from the American College of Radiology's 1995 Survey of Radiologists. Radiation oncologists, the retired, and residents and fellows were excluded. Responses were weighted to represent all practicing diagnostic radiologists in the United States and were compared with a similar previous survey. RESULTS: More than one quarter (28%) of diagnostic radiologists are subspecialists. Diagnostic radiology subspecialists were more likely than generalists to have fellowship training, work in center cities, work in large groups, and be employed by an academic institution. They were also generally younger and provided a narrower range of imaging services than generalists. CONCLUSION: Although most diagnostic radiologists are generalists, subspecialization will likely continue to grow. PMID- 10789765 TI - Malpractice issues in radiology. When an expert witness is not an expert. PMID- 10789764 TI - Diagnostic radiology residents in the classes of 1999 and 2000: fellowship and employment. AB - OBJECTIVE: Most diagnostic radiology residents undergo fellowship training. Applications for fellowship positions are submitted during the third year of residency. We compared the number of residents undertaking fellowship, accepting jobs, and rescinding accepted fellowship positions to accept jobs. MATERIALS AND METHODS: During the 47th Association of University Radiologists meeting, participants from the American Association of Academic Chief Residents in Radiology completed a questionnaire regarding the intentions of third- and fourth year residents in their programs. RESULTS: Seventy programs, representing 402 fourth-year and 395 third-year residents, responded. Of fourth-year residents, 322 (80.1%) accepted fellowships and 41 (10.2%) accepted jobs. Of third-year residents, 335 (84.8%) accepted fellowships. Eighty-eight (27.3%) fourth-year residents and 95 third-year residents (28.4%) accepted interventional radiology fellowships, and 104 (32.3%) fourth-year residents and 96 (28.7%) third-year residents accepted body imaging fellowships. Most residents who accepted fellowships did so in the same city in which they completed their residency training. Of fourth-year residents who accepted fellowship positions the year before, 14 (4.3%) withdrew their acceptance to pursue employment. CONCLUSION: Most residents opt for fellowship training in interventional radiology or body imaging. We postulate that the practice of accepting fellowships during the third year of residency may contribute to the percentage of residents who rescind fellowship acceptance to pursue employment. We suggest that consideration be given to changing this practice. PMID- 10789766 TI - Physician training requirements in sonography: how many cases are needed for competence? AB - OBJECTIVE: Physician competence in the performance of sonographic studies was assessed after their involvement in predetermined increments of cases to determine whether the case volumes currently required by the American Institute of Ultrasound in Medicine and the American College of Radiology for training in sonography can be lowered substantially. MATERIALS AND METHODS: Sonographic competence tests were administered to 10 first-year diagnostic radiology residents after their involvement in increments of 50 cases, up to a total of 200 cases (four competency tests). Each competency test consisted of the resident's independently scanning and interpreting 10 clinically mandated studies that were scored in comparison with the examination performed by the sonographer and interpreted by an attending radiologist. Trainee studies were graded on the percentage of anatomic landmarks depicted, the number of reporting errors, the number of clinically significant reporting errors, and the percentage of cases receiving a passing score. RESULTS: Although resident performance improved progressively with increasing experience for all parameters assessed, performance of the group was poor even after their involvement in 200 cases. At this testing level, the mean percentage of anatomic landmarks depicted successfully was 56.5%; the mean total reporting errors per case was 1.2; the mean clinically significant errors per case was 0.5; and the mean percentage of cases receiving a passing score was 16%. Impressive performance differences were observed among residents for all parameters assessed, and these differences were not explained by the number of months of radiology training the resident had taken before the sonography rotation. CONCLUSION: Involvement in 200 or fewer cases during the training period is not sufficient for physicians to develop an acceptable level of competence in sonography. PMID- 10789767 TI - On improvement of scientific presentations. AB - Scientific presentations require a great deal of preparation. Background lettering colors on the slide, the number of words on a slide, and the elimination of distracting factors are all important in determining the success of one's presentation. This brief paper has attempted to address some of these issues. PMID- 10789768 TI - The Spanish-American War and military radiology. PMID- 10789769 TI - A simple method for obtaining original data from published graphs and plots. AB - OBJECTIVE: To describe a method for deriving original data values from scanned images of graphs and scatterplots published in the medical literature. CONCLUSION: The procedure is simple, reproducible, and relatively error free (when performed carefully). This method is useful in converting published graphic material into numeric data for various uses when the original data are unavailable directly from the authors. PMID- 10789770 TI - Core needle biopsy of challenging benign breast conditions: a comprehensive literature review. PMID- 10789771 TI - Effect of obesity on screening mammography: outcomes analysis of 88,346 consecutive examinations. AB - OBJECTIVE: We determined differences in the rates of recall, biopsy, and cancer detection for screening mammography as a function of adiposity. MATERIALS AND METHODS: Eighty-eight thousand three hundred forty-six consecutive screening mammography examinations were performed from April 1985 to August 1997. Patient weights were normalized to ideal weight correcting for height and were subdivided into adiposity cohorts including underweight by greater than 10%; ideal weight +/ 10%; overweight by 11-24%; overweight by 25-39%; and overweight by greater than 40%. The rates of recall, biopsy, cancer detection, and cancer stage were calculated and were analyzed using the chi-square test for trend. Cancer size was analyzed using linear regression analysis. RESULTS: Reliable (p < 0.05) and meaningful differences were seen between cohorts of increasing adiposity for rates of recall, biopsy, and cancer detection. An increase in recall rate occurred with progressively increasing adiposity (3.88%, 4.89%, 5.11%, 5.47%, 5.55% [p < 0.0001]). The rate of biopsy also increased with increasing adiposity (0.98%, 1.31%, 1.35%, 1.59%, 1.65% [p < 0.0002]), as did the rate of screening detected cancer (number of cases of cancer per 1000 women screened) (3.74, 4.29, 5.34, 4.70, 6.04 [p < 0.015]). Finally, increased adiposity also correlated with increased median cancer size (p < 0.02) and with more advanced stage at diagnosis (p = 0.046). CONCLUSION: Increasing adiposity correlates with progressive increases in the rates of recall, biopsy, and cancer detection for women undergoing screening mammography. Increasing adiposity also correlates with increased cancer size and stage, providing further support for obesity as a risk factor for breast cancer. PMID- 10789772 TI - Centennial photo page. From the AJR archives: celebrating the ARRS centennial. Fifty-three cases of carcinoma of the breast: occult until mammography. PMID- 10789773 TI - Accuracy of screening mammography using single versus independent double interpretation. AB - OBJECTIVE: We conducted an analysis among 31 community radiologists to identify the average change in screening mammography interpretive accuracy afforded by independent double interpretation. MATERIALS AND METHODS: We assessed interpretive accuracy using a stratified random sample of test mammograms that included 30 women with cancer and 83 without. Radiologists were unaware of clinical information and of each other's assessments. We describe accuracy for individual radiologists and for double interpretation, including average sensitivity, specificity, diagnostic likelihood ratios positive and negative, and area under the receiver operating characteristic (ROC) curve. We also assessed weighted and nonweighted kappa statistics among all 465 pairs of radiologists and 31,465 pairs of unique pairs. The assessment for double interpretations used the "highest" (i.e., most abnormal) assessment of the two radiologists. We calculated the difference between each radiologist's individual accuracy and the average accuracy across that radiologist's 30 double interpretations. RESULTS: We found the following average accuracy statistics for individual radiologists: sensitivity, 79%; specificity, 81%; diagnostic likelihood ratio positive, 5.53; diagnostic likelihood ratio negative, 0.26; and area under the ROC curve, 0.85. The mean kappa statistic among radiologists for cancer cases increased with double interpretation from 0.59 to 0.70, and for noncancer cases from 0.30 to 0.34. Double interpretation resulted in an average increase in sensitivity of 7%, an average decrease in specificity of 11%, a decrease in diagnostic likelihood ratio positive of 2.35, a decrease in diagnostic likelihood ratio negative of 0.06, and an increase in area under the ROC curve of 0.02. CONCLUSION: Independent double interpretation does not increase accuracy as measured by the area under the ROC curve. PMID- 10789774 TI - Incidental treatment of nipple discharge caused by benign intraductal papilloma through diagnostic Mammotome biopsy. AB - OBJECTIVE: The purpose of this study was to evaluate imaging-guided vacuum assisted mammotome biopsy as a minimally invasive method of obtaining a satisfactory diagnosis and eliminating the bothersome symptoms in patients presenting with nipple discharge. MATERIALS AND METHODS: Forty-nine women who presented with nipple discharge and who had final pathologic diagnoses of papillary lesions were retrospectively identified. Fifty-six lesions were biopsied in this group. The examinations included mammography, ductography, sonography, and, if possible, percutaneous biopsy. All lesions were centrally located and most were superficial. Of this study group, four patients with five lesions proceeded to sonographically guided automated core biopsy, and 38 patients with 44 intraductal lesions identified by sonography advanced to sonographically guided biopsy with an 11-gauge mammotome probe. One patient underwent stereotactic 11-gauge mammotome biopsy. Patients not advancing to sonographically guided biopsy were those with masses either in the nipple or nipple-areolar complex (five patients), one patient with no identifiable lesion at sonography, and one directly referred for open surgical biopsy. RESULTS: In all biopsied patients, satisfactory tissue for diagnosis was obtained. In patients biopsied with the mammotome probe, follow-up at a mean time of 13 months revealed resolution of the presenting problematic discharge in 97.2% of patients. Complications were mild and infrequent. Only one of 50 percutaneously biopsied lesions was not benign and required subsequent surgery. CONCLUSION: Papilloma excision with percutaneous biopsy allows safe and accurate tissue analysis and a high probability of terminating the symptomatic nipple discharge. PMID- 10789775 TI - Noninvasive imaging of cervical vascular injuries. AB - Various imaging examinations can be used to diagnose cervical vascular injuries. The challenge in the current medical environment is to choose the imaging examination that is the most rapid, least invasive, and least costly. One must recognize the ability of each technique to detect lesions, taking into consideration the type of abnormality and whether the carotid or vertebral arteries are at risk for injury. PMID- 10789776 TI - Contrast-enhanced color Doppler sonography for evaluation of enlarged cervical lymph nodes in head and neck tumors. AB - OBJECTIVE: We determined whether contrast-enhanced color Doppler sonography can differentiate benign from malignant enlarged cervical lymph nodes in head and neck tumors. SUBJECTS AND METHODS: Ninety-four enlarged lymph nodes in 39 adult patients (32 men and seven women; age range, 30-81 years) were examined with B mode sonography and with unenhanced and contrast-enhanced color Doppler sonography. All patients had carcinoma of the oral cavity. Histologically, lymphadenitis was found in 57 nodes and metastases in 37 nodes. Geometric dimension, texture, and margin of the node and detection and location of vessels were noted. Histology and imaging findings were correlated. RESULTS: The transverse-to-longitudinal diameter ratio in combination with texture and margin analysis resulted in a correct diagnosis in only approximately 79% of the nodes. With contrast-enhanced color Doppler sonography, 86% of nodes showed vessels, and 28% of nodes showed vessels with this technique exclusively. Characteristic configurations were identified: hilar vessels with branching indicated lymphadenitis (sensitivity, 98%; specificity, 100%), and predominantly peripheral vessels indicated metastases (100%, 98%). These findings changed the diagnosis in 13 nodes, changed the therapy in four patients, and led to an incorrect diagnosis in one patient. CONCLUSION: Enlarged lymph nodes can be characterized as metastatic or inflammatory with high diagnostic accuracy on the basis of their vascular architecture as seen on contrast-enhanced color Doppler sonography. PMID- 10789777 TI - Laryngeal rhabdomyoma involving the paraglottic space. PMID- 10789778 TI - Radiation-induced osteosarcoma after bilateral childhood retinoblastoma. PMID- 10789779 TI - Sinus lift procedure of the maxilla in patients with inadequate bone for dental implants: radiographic appearance. AB - OBJECTIVE: Dental implants have gained popularity for treating edentulism, but some patients develop jaw atrophy, which leaves insufficient bone for implants. To treat these patients, the sinus lift procedure, which augments bone, was developed. Altered anatomy from this procedure has an unusual radiographic appearance, confusing those unfamiliar with it. We describe the sinus lift procedure and its radiographic appearance. CONCLUSION: With knowledge of this surgery and some of its pitfalls, radiographs can be more easily and accurately interpreted. PMID- 10789780 TI - Cerebral dural arteriovenous fistulas: detection by dynamic MR projection angiography. PMID- 10789781 TI - Traumatic ossicular disruption. PMID- 10789782 TI - A guide to the identification of major cerebral arteries with transcranial color Doppler sonography. PMID- 10789783 TI - Early diffusion-weighted MR imaging abnormalities in sustained seizure activity. PMID- 10789784 TI - Benign abnormalities and carcinoid tumors of the central airways: diagnostic impact of CT bronchography. AB - OBJECTIVE: The purpose of this retrospective study was to determine the added diagnostic value, if any, of CT bronchography for the detection and characterization of benign abnormalities and typical carcinoid tumors of the central airways. MATERIALS AND METHODS: We used bronchoscopy and helical CT to examine 238 bronchial sections in 28 patients with 32 bronchial abnormalities and in five patients with normal bronchoscopy results. Postprocessing consisted of CT bronchography based on surface rendering. Images were interpreted independently by two observers (a radiologist and a pneumonologist) who were not informed of the bronchoscopy results. After initial interpretation of axial CT scans, the observers analyzed the axial CT scans with CT bronchograms. Results were evaluated for gain in diagnostic accuracy and in confidence. RESULTS: Mean sensitivity for detection of abnormal bronchial sections was 89% (range, 87-90%) for axial CT and 92% (range, 90-94%) for axial CT with CT bronchography (not significant). Mean specificity of both approaches exceeded 99%. A correct diagnosis of the nature of the bronchial abnormalities was proposed for 68% of the cases in which axial CT was used alone and in 76% in which both axial CT and CT bronchography were used (not significant). The addition of CT bronchography significantly increased the confidence of the pneumonologist in the diagnoses. CONCLUSION: Axial CT remains the technique of choice to detect and characterize benign abnormalities of the airways. CT bronchography provides little diagnostic gain but increases the confidence of chest physicians in the interpretation of CT scans for the assessment of benign abnormalities and typical carcinoids of the central airways. PMID- 10789785 TI - Using CT to diagnose nonneoplastic tracheal abnormalities: appearance of the tracheal wall. PMID- 10789786 TI - Pulmonary sequelae of bronchopulmonary dysplasia survivors: high-resolution CT findings. AB - OBJECTIVE. We examined the high-resolution CT findings of adult survivors of bronchopulmonary dysplasia. CONCLUSION. The cardinal CT features of bronchopulmonary dysplasia survivors include multifocal areas of reduced lung attenuation and perfusion, bronchial wall thickening, and decreased bronchus-to pulmonary artery diameter ratios. PMID- 10789787 TI - Pulmonary involvement with Erdheim-Chester disease: radiographic and CT findings. AB - OBJECTIVE: We determined the chest radiographic and CT findings of Erdheim Chester disease with pulmonary involvement. MATERIALS AND METHODS: We retrospectively reviewed the radiologic images of 15 patients with biopsy-proven Erdheim-Chester disease. Nine patients had chest radiographic images and CT scans that were available for review. Six men and three women were studied (age range, 25-70 years; mean age, 56 years). Two radiologists interpreted all images by consensus. Lung parenchyma was assessed for the type and distribution of disease. Bronchi, pleurae, hila, and mediastinal and extrathoracic structures were evaluated for abnormalities. Pathologic specimens from all patients were reviewed and correlated with radiologic findings. RESULTS: Eight of nine patients had thoracic images with abnormal findings. The most common radiographic pattern was reticular interstitial opacities with fissural and interlobular septal thickening. CT revealed regions of ground-glass attenuation and centrilobular nodular opacities. Typically, extrapulmonary findings included pleural effusions (6/8 patients), pericardial fluid or thickening (4/8), and extrathoracic infiltrative soft-tissue masses (4/8). CONCLUSION: The most common findings of Erdheim-Chester disease with pulmonary involvement include an interstitial process characterized by smooth interlobular septal thickening and centrilobular nodular opacities, fissural thickening, and pleural effusions. On CT, six of nine patients had pericardial fluid and thickening or extrathoracic soft-tissue masses. Such findings are characteristic of Erdheim-Chester disease with pulmonary involvement. Definitive diagnosis requires correlating skeletal findings and lung biopsy findings. PMID- 10789789 TI - Chest radiographic diagnosis of stomach abnormality. PMID- 10789788 TI - Creation of a three-dimensional model of human segmental lung anatomy. AB - OBJECTIVE: The investigation of pulmonary embolism using scintigraphic tomography requires a model of the internal architecture of the segments and subsegments in the human lung. Such a model has been developed by the segmentation and subsegmentation of an existing whole-body tissue-segmented phantom. MATERIALS AND METHODS: By using information from suitably windowed human axial CT scans, combined with the information gained from the injection of color-coded dyes into the segmental bronchi of human cadaveric lungs, the lobar and segmental boundaries were added to the existing phantom. Further refinements were added from reports in the literature regarding the predominant pattern of subsegmental bronchi in a series of human cadavers, enabling the creation of subsegmental boundaries. RESULTS: A digitized model of the segmental and subsegmental anatomy of the human lung was successfully created. External, or pleural, projections of the complex internal arrangement of the segments closely corresponded with the projections of the best available authorities on the subject. CONCLUSION: The model provides the opportunity to address several issues germane to scintigraphy and important for diagnosing pulmonary embolic disease. In particular, the model allows the manipulation of three-dimensional data sets to explore issues of importance to tomographic lung scanning. PMID- 10789790 TI - Cardiovascular instability caused by inadvertent positive end-expiratory pressure in a patient with panlobular emphysema receiving mechanical ventilation: radiographic-physiologic correlation. PMID- 10789791 TI - Lung torsion after lung transplantation: evaluation with helical CT. PMID- 10789792 TI - Medicine in American art. Dr. Peter Guernsey. PMID- 10789793 TI - Virtual endoscopy of coronary arteries using contrast-enhanced ECG-triggered electron beam CT data sets. PMID- 10789794 TI - Causes of transient dilatation of the left ventricle during myocardial perfusion imaging. PMID- 10789795 TI - Using sonography to reveal and aspirate joint effusions. PMID- 10789796 TI - Subchondral marrow changes after laser diskectomy in the lumbar spine: MR imaging findings and clinical correlation. AB - OBJECTIVE: We examined MR imaging findings and determined the clinical significance of subchondral bone marrow changes in the lumbar spine of holdium:yttrium aluminum garnet laser diskectomy patients. MATERIALS AND METHODS: We retrospectively reviewed the pre- and postoperative MR images of 109 patients with recurrent radiculopathy, lower back pain, or both 1 year after laser diskectomy of 178 disks. From this group of patients, MR images were also obtained in 11 patients with subchondral marrow changes 5-7 years after surgery. These patients were interviewed regarding residual lower back pain. Thirteen asymptomatic laser diskectomy patients also underwent follow-up MR imaging within 1 year of surgery. RESULTS: After surgery, subchondral marrow abnormalities were identified in 41 of 109 laser diskectomy patients. The remaining 68 patients had no postoperative subchondral bone marrow changes. Postoperative subchondral marrow changes were not associated with inflammation of the adjacent disk space and did not affect surgical outcome. Bone marrow changes decreased in size in the 11 patients examined 5-7 years after laser diskectomy, and eight of these patients described their lower back pain as improved. In 13 asymptomatic laser diskectomy patients, one new subchondral marrow abnormality was identified. CONCLUSION: Subchondral marrow abnormalities may occur in the vertebral end plates after holmium:yttrium aluminum garnet laser diskectomy. However, these changes probably do not affect surgical outcomes and appear to resolve over time. PMID- 10789797 TI - MR imaging assessment of the pectoralis major myotendinous unit: an MR imaging anatomic correlative study with surgical correlation. AB - OBJECTIVE: MR imaging is the optimal imaging technique to study the normal and abnormal conditions of the pectoralis major muscle and tendon unit. The purpose of this study was to use MR imaging to provide an anatomic survey of the normal pectoralis major tendon and its insertion and to compare these findings with surgically proven cases of rupture. CONCLUSION: MR imaging shows the normal pectoralis major myotendinous unit has low signal intensity on both T1- and T2 weighted images. Reliable anatomic landmarks for visualization and examination of injuries to the muscle and myotendinous unit include the quadrilateral space, or the origin of the lateral head of the triceps, as the superior boundary and the deltoid tuberosity as the inferior boundary of the intact tendon of insertion. Failure to visualize a normal insertion within these boundaries should prompt a dedicated search by the radiologist for rupture and retraction of the tendon medially. PMID- 10789798 TI - MR anatomy of the subcoracoid bursa and the association of subcoracoid effusion with tears of the anterior rotator cuff and the rotator interval. AB - OBJECTIVE: We describe the MR anatomy of the subcoracoid bursa and findings associated with subcoracoid effusion. CONCLUSION: Fluid in the subcoracoid space, revealed on MR imaging of the shoulder, may lie in the subcoracoid bursa or the subscapularis recess (both structures can be distinguished with MR imaging). In our patients, subcoracoid effusions were often associated with anterior rotator cuff tears, including tears of the rotator interval. PMID- 10789799 TI - Association of paralabral cysts with acetabular disorders. AB - OBJECTIVE: Acetabular labral tears may cause considerable pain and predispose patients to premature osteoarthritis. Therefore, early and accurate examination is required. Acetabular labral tears are difficult to view on MR images. We report the association of paralabral ganglion cysts detected on MR imaging with surgically proven acetabular labral disorders. CONCLUSION: Paralabral cysts may be associated with labral disorders. The appearance of a paralabral cyst on MR imaging is a useful indirect sign of acetabular labral abnormality. Use of this sign may enable earlier and more accurate examination of labral disorders. PMID- 10789800 TI - Dynamic contrast-enhanced MR imaging of bacterial abscess and VX2 carcinoma in rabbits: comparison of gadopentetate dimeglumine and a macromolecular contrast agent. AB - OBJECTIVE: The objective of this study was to compare enhancement patterns of a blood-pool contrast agent, Gadomer-17, with those of gadopentetate dimeglumine in bacterial abscesses and VX2 carcinoma in rabbits. MATERIALS AND METHODS: Fourteen rabbits with experimentally induced bacterial abscesses and VX2 carcinoma in both thighs underwent dynamic contrast-enhanced MR imaging with Gadomer-17 and gadopentetate dimeglumine at a 24-hr interval. The enhancement ratios (postcontrast to precontrast signal intensities) of lesions in the same animal were assessed and correlated with microvessel density. RESULTS: For Gadomer-17, the enhancement ratio of the abscesses (1.66 +/- 0.39) peaked 15 min after the injection, while that of the carcinoma (2.05 +/- 0.16) peaked at 10 min. The enhancement ratios of the carcinoma were consistently higher than those of the abscesses up to 30 min. For gadopentetate dimeglumine, peak enhancement ratio of the abscesses (2.30 +/- 0.75) was seen 5 min after the injection, while that of the carcinoma (2.32 +/- 0.51) was seen at 3 min. The enhancement ratios of the carcinomas were significantly higher at 1 min, but significantly lower at 20-30 min, compared with those of the abscesses, as a result of rapid decrease of enhancement ratios in the carcinomas. The microvessel density was 9.8 +/- 5.2 vessels per field of view for the abscesses and 36.3 +/- 9.5 vessels per field of view for the carcinoma (p < 0.001). CONCLUSION: Delayed peak enhancement and slow decay were found in both bacterial abscess and VX2 carcinoma with Gadomer-17, whereas early peak enhancement and rapid decay were found especially in VX2 carcinoma with gadopentetate dimeglumine. Enhancement ratios on MR imaging with a blood-pool contrast agent correlated well with the microvessel density in bacterial abscess and VX2 carcinoma. PMID- 10789801 TI - Impact of abdominal CT on the management of patients presenting to the emergency department with acute abdominal pain. AB - OBJECTIVE: The purpose of this study is to document the impact of CT performed in the emergency department of patients presenting with nontraumatic acute abdominal pain. SUBJECTS AND METHODS: Fifty-seven patients were enrolled in this prospective study. Using a computer order entry system, emergency department physicians were required to report their most likely diagnosis, level of certainty, and management plan for their patients before ordering abdominal CT. After CT was performed, each physician was required to provide again his or her diagnosis, level of diagnostic certainty, and treatment plan. The outcome of each patient was evaluated by either surgery, other imaging studies, or clinical follow-up. RESULTS: After the abdominal CT, physicians' mean level of certainty in their diagnoses increased by 1.5 points (on a five-point scale; p < 0.0001). Patient management was changed in 33 (60.0%) of 55 patients. Planned treatment before CT was admission in 42 patients. Actual admissions after CT totaled 32 patients (excluding the two patients in whom preimaging information was not recorded). Thus, the net effect of abdominal CT scanning was to avert 10 (23.8%) of 42 hospital admissions. CONCLUSION: CT performed in the emergency department increases the physician's level of certainty, reduces hospital admission rates by 23.8%, and leads to more timely surgical intervention. PMID- 10789802 TI - Acute diverticulitis of the cecum and ascending colon: the value of thin-section helical CT findings in excluding colonic carcinoma. AB - OBJECTIVE: The purpose of this study was to assess the value of characteristic thin-section helical CT findings of acute diverticulitis involving the cecum and ascending colon in excluding colonic carcinoma. MATERIALS AND METHODS: Thin section helical CT scans (5-mm collimation) of 19 consecutive patients with proven diverticulitis and 21 consecutive patients with surgically proven carcinoma involving the cecum and ascending colon were reviewed retrospectively. Two radiologists independently analyzed these parameters: degree of pericolic infiltration, mesenteric fluid, vascular engorgement, arrowhead-shaped wall thickening, air-filled diverticula, inflamed diverticula, and preserved enhancement pattern of involved colonic wall. Inter-observer agreement was assessed with a kappa statistical analysis, and the features that most distinguished diverticulitis from colonic carcinoma were selected with a stepwise logistic-regression analysis. RESULTS: The two CT findings of right-sided colonic diverticulitis that most distinguished it from colonic carcinoma were inflamed diverticula and the preservation of an enhancement pattern of the involved colonic wall. Excellent interobserver agreement (kappa > 0.60) was obtained for both findings. Inflamed diverticula (kappa = 0.80) had a mean sensitivity, specificity, and accuracy for diverticulitis of 86.8%, 92.9%, and 90.0%, respectively, in differentiating right-sided colonic diverticulitis from colonic carcinoma. Preserved wall enhancement pattern (kappa = 0.70) had a mean sensitivity, specificity, and accuracy of 89.5%, 95.3%, and 92.5%, respectively. CONCLUSION: On thin-section helical CT, an inflamed diverticula and a preserved enhancement pattern of the thickened colonic wall were the two most statistically significant CT findings of acute diverticulitis involving the cecum and ascending colon that distinguished diverticulitis from colonic carcinoma. PMID- 10789803 TI - MR cholangiopancreatographic differentiation of benign and malignant intraductal mucin-producing tumors of the pancreas. AB - OBJECTIVE: The purpose of this study was to establish MR cholangiopancreatographic criteria for discriminating benign from malignant intraductal mucin-producing tumors of the pancreas. MATERIALS AND METHODS: Thirty one patients with 34 intraductal mucin-producing tumors underwent MR cholangiopancreatography. Tumors were classified as either main duct type (n = 10) or branch duct type (n = 24). In patients with the main duct type, the maximum diameter and the location of the main pancreatic duct, the extent of main pancreatic duct dilatation, and the presence of a filling defect were evaluated. For branch duct type, the location and maximum diameter of the cystic lesion, the presence of a filling defect, and the presence of associated main pancreatic duct dilatation were evaluated. RESULTS: In patients with the main duct type, the main pancreatic duct was significantly narrower when associated with benign rather than malignant tumors. All malignant tumors showed diffuse main pancreatic duct dilatation, whereas all benign tumors showed segmental dilatation. Among patients with branch duct type, the cyst was smaller when it was a benign rather than malignant tumor. All but one malignant tumor showed mild associated main pancreatic duct dilatation, whereas benign tumors were not associated with main pancreatic duct dilatation. Filling defects suggested malignancy, although half of the malignant tumors had no filling defects. CONCLUSION: In patients with intraductal mucin-producing tumors of the pancreas, filling defects are indicative of malignancy. Diffuse main pancreatic duct dilatation greater than 15 mm (main duct type), or any main pancreatic duct dilatation (branch duct type), is strongly associated with malignancy. PMID- 10789804 TI - Videofluoroscopy of swallowing in symptomatic patients who have undergone long term intubation. AB - OBJECTIVE: The aim of this study was to evaluate whether specific patterns of swallowing dysfunction occur in symptomatic patients after long-term intubation. SUBJECTS AND METHODS: Twenty-one patients (16 men, five women; mean age, 66 years) who presented with clinical signs of aspiration after long-term intubation (mean duration, 24.6 days) underwent videofluoroscopy. They were analyzed for functional abnormalities of the tongue, soft palate, epiglottis, hyoid and larynx, pharynx, and the upper esophageal sphincter. We assessed the presence or absence of aspiration, the type of aspiration (pre-, intra-, and postdeglutitive), and a spectrum of other swallowing abnormalities. RESULTS: There were 18 patients (86%) with radiologically proven aspiration. In another patient only laryngeal penetration occurred. There were 11 combinations of pre-, intra-, and postdeglutitive aspiration. Predeglutitive aspiration was predominant and present in 52% of our patients. We found functional abnormalities of the tongue in 48%, of the soft palate in 10%, of the epiglottis in 48%, of the pharynx in 71%, and of the upper esophageal sphincter in 24%. CONCLUSION: Patients who are symptomatic after undergoing long-term intubation do not develop a specific type or pattern of swallowing dysfunction or aspiration, but show a large variety of aspiration types and associated swallowing disorders. Nevertheless, videofluoroscopy has the ability to reveal complex deglutition disorders and to aid precise planning of individualized functional swallowing therapy. PMID- 10789805 TI - Using CT to diagnose traumatic lumbar hernia. AB - OBJECTIVE: The objective of this study was to determine the CT findings of traumatic lumbar hernia in 15 patients and to discuss the mechanism and treatment of injury. CONCLUSION: CT can reveal traumatic lumbar hernia and show both the anatomy of disrupted muscular layers and the presence of herniated intraabdominal viscera or retroperitoneal fat. PMID- 10789806 TI - Dual phase hepatic CT: influence of scanning direction on liver attenuation. AB - OBJECTIVE: We measured changes in hepatic attenuation during arterial and portal phase acquisition of hepatic CT in the craniocaudal and caudocranial directions. SUBJECTS AND METHODS: In 10 of 20 patients undergoing dual phase helical CT during staging for colorectal cancer, images in both phases were obtained in the craniocaudal direction. Ten patients underwent imaging in the caudocranial direction. Attenuation values in the aorta and in the peripheral and central liver regions of interest were measured on each slice. Central and peripheral liver attenuation was also measured in 10 additional patients undergoing unenhanced CT. RESULTS: Both peripheral and central regions of interest revealed progressively increasing attenuation during the arterial phase, irrespective of scanning direction. During the portal phase, hepatic attenuation was stable in the craniocaudal direction but decreased in the caudocranial direction (p < 0.05, Wilcoxon's signed rank sum test). Central hepatic attenuation was lower than peripheral attenuation in unenhanced livers and in enhanced livers during both phases of caudocranial acquisition. We determined no significant difference during the arterial phase of enhancement in the craniocaudal direction. CONCLUSION: The direction of acquisition does not influence sequential liver enhancement during the arterial phase. Craniocaudal acquisition produces more stable enhancement during the portal phase. Differences in attenuation between the central and peripheral areas of the liver are probably unrelated to contrast administration. PMID- 10789807 TI - Single breath-hold T2-weighted MR imaging of the liver: value of single-shot fast spin-echo and multishot spin-echo echoplanar imaging. AB - OBJECTIVE: The objective of our study was to evaluate the efficacy of single breath-hold T2-weighted MR imaging for detection of focal hepatic lesions. MATERIALS AND METHODS: T2-weighted MR images were retrospectively reviewed from 51 patients with 85 solid and 59 nonsolid lesions using the following four sequences: conventional spin-echo, respiratory-triggered fast spin-echo, single shot fast spin-echo, and multishot spin-echo echoplanar imaging. Images were reviewed on a hepatic segment-by-segment basis; T2-weighted images of a total of 408 hepatic segments were reviewed separately and independently for solid and nonsolid lesions by four radiologists. Quantitative, qualitative, and receiver operating characteristic analyses were performed. RESULTS: For solid lesions, no significant differences were seen among the lesion-to-liver contrast-to-noise ratios with the four sequences. In terms of solid lesion detection, no significant difference was seen between the diagnostic accuracy of multishot spin echo echoplanar (Az = 0.90) and respiratory-triggered fast spin-echo (Az = 0.91) imaging, which showed the best performance of the four sequences. For nonsolid lesion detection, respiratory-triggered fast spin-echo and single-shot fast spin echo imaging were judged the best (Az = 0.94). CONCLUSION: Breath-hold single shot fast spin-echo and multishot spin-echo echoplanar sequences can be substituted for conventional spin-echo and respiratory-triggered fast spin-echo T2-weighted sequences. PMID- 10789808 TI - Sonography for selecting candidates for laparoscopic cholecystectomy: a prospective study. AB - OBJECTIVE: We assessed the value of sonography in predicting intraoperative difficulties for patients undergoing laparoscopic cholecystectomy and in identifying indicators for conversion to conventional cholecystectomy. SUBJECTS AND METHODS: Upper abdominal sonography was performed (according to a checklist) in 75 consecutive patients before laparoscopic cholecystectomy. Sonographic findings were verified by the surgeon in the operating room. RESULTS: Conversion from laparoscopic surgery to laparotomy was performed in five patients (6.7%). Of 75 patients, 19 had sonograms revealing gallbladder wall thickening (>4 mm); surgical preparation difficulties in 16 of these patients led to laparotomy in four patients. Sensitivity, specificity, positive predictive value, and accuracy of wall thickening as an indicator of technical difficulties were 66.7%, 94.1%, 84.2%, and 85.3%, respectively. Sensitivity, specificity, positive predictive value, and accuracy of wall thickening as an indicator of surgical conversion were 80.0%, 78.6%, 21.1%, and 78.7%, respectively. Technical difficulties at laparoscopy occurred in all five patients with pericholecystic fluid on sonography (sensitivity, 20.8%; specificity, 100%; positive predictive value, 100%; accuracy, 74.7%) and led to laparotomy in three patients (sensitivity 60.0%, specificity 97.1%, positive predictive value 60%, accuracy 94.7%). The accuracy of sonography for cholecystolithiasis was 100%. CONCLUSION: On sonography, gallbladder wall thickening is the most sensitive indicator and pericholecystic fluid is the most specific indicator of technical difficulties during laparoscopic cholecystectomy. Such difficulties may require conversion to laparotomy. PMID- 10789809 TI - Radiologic features of complications arising from dropped gallstones in laparoscopic cholecystectomy patients. AB - OBJECTIVE: Because laparoscopic cholecystectomy has become the accepted treatment for symptomatic cholelithiasis, radiologists frequently encounter patients who have had this surgery. Although the radiologic features of postoperative bile duct injury are well documented, the imaging features of less well-known complications remain poorly described. One such unusual complication is abscess formation caused by dropped gallstones. CONCLUSION: Although the incidence of dropped gallstones is an uncommon complication of laparoscopic cholecystectomy, it should be recognized as a potential source of both intraabdominal and intrathoracic abscess formation in any patient presenting months to years after undergoing laparoscopic cholecystectomy. These abscesses are not necessarily confined to the right upper quadrant. PMID- 10789810 TI - Three-dimensional gadolinium-enhanced MR angiography of vascular complications after liver transplantation. AB - OBJECTIVE: The goal of this study was to evaluate three-dimensional gadolinium enhanced MR angiography as a tool for examination of liver transplant patients with potential vascular complications. MATERIALS AND METHODS: Thirty-eight consecutive three-dimensional gadolinium-enhanced MR angiograms were obtained in 34 patients. Results were retrospectively reviewed and correlated with conventional angiography in 20 of the 38 cases and sonography in 37 of the 38 cases. MR angiograms were evaluated for technical adequacy, vascular patency, and parenchymal abnormalities, and results were compared with angiography and sonography. Conventional angiography and surgery were used as gold standards when available. RESULTS: Thirty-four (90%) of 38 MR angiograms were technically adequate. Vascular abnormalities were identified in 20 patients, and 19 of these patients subsequently underwent angiography, surgery, or both. There were seven cases of hepatic artery thrombosis; all were detected with MR angiography with no false-positive or false-negative interpretations. Seven patients had moderate to severe hepatic artery stenosis (>50% narrowing as determined by conventional angiography). MR angiography revealed this stenosis in six of the seven patients, with one false-negative and three false-positive interpretations. Portal vein thrombosis was detected in three patients, and portal vein stenosis was detected in two patients. CONCLUSION: Three-dimensional gadolinium-enhanced MR angiography is useful in the examination of liver transplant patients and offers a noninvasive adjunct in patients with difficult or indeterminate sonographic examinations. PMID- 10789811 TI - Effectiveness and safety of balloon dilation of the papilla and the use of an occlusion balloon for clearance of bile duct calculi. AB - OBJECTIVE: The purpose of this study was to describe a technique for percutaneous bile duct stone clearance by pushing the stones into the small bowel after balloon dilation of the papilla. SUBJECTS AND METHODS: During a 2-year period, 38 patients were treated percutaneously for stones in the biliary tree. Twenty-one patients were treated through a T tube or transcystic tract. Seventeen patients were treated through a transhepatic tract. Twenty-three patients had one stone each. Eight patients had two stones, and seven patients had three or more calculi. Stone size ranged from 3 to 16 mm in diameter (mean size, 6.7 nm). Balloon diameter based on the transverse diameter of the stones ranged from 7 to 18 mm (mean, 6.7 mm). An 11.5-mm occlusion balloon was used for pushing the stones through a 7- to 9-French vascular introducer. A catheter was left in the common bile duct from 1 to 6 days for external drainage. RESULTS: The technique was successfully used for clearance of stones in 36 (94.7%) of the 38 patients. With 29 patients, the procedure was performed with only one attempt. Two attempts were necessary for five patients, and three attempts were necessary for four patients. Two major complications were cholangitis and biliary pleural effusion. No deaths were related to the procedure. CONCLUSION: Percutaneous bile duct stone clearance by dilation of the papilla and evacuation of the stones in an antegrade fashion with an occlusion balloon is a safe and effective technique. It can be an alternative to basketing stones in selected patients. PMID- 10789812 TI - Do women with palpable masses need bilateral mammography if they have already had a mammogram in the last 12 months or are under 40? PMID- 10789813 TI - On missed breast cancer. PMID- 10789814 TI - Megapixel digital camera. PMID- 10789815 TI - Radiology lexicon. PMID- 10789816 TI - Color Doppler sonography of herniated paraumbilical collateral vein masquerading as an acquired umbilical hernia. PMID- 10789817 TI - Milk of calcium in Meckel's diverticulum. PMID- 10789818 TI - CT of calcific cerebral emboli after carotid manipulation. PMID- 10789819 TI - Duchenne-Landouzy dystrophy: CT explanation of low serum creatinine level. PMID- 10789820 TI - CT of tattoos removed with laser therapy. PMID- 10789821 TI - Cardiac mucormycosis revealed on imaging. PMID- 10789822 TI - Dobutamine stress echocardiography: safety in diagnosing coronary artery disease. AB - Dobutamine stress echocardiography is considered a relatively well-tolerated diagnostic modality, effective in the management of patients with known or suspected coronary artery disease. Adverse effects during testing are relatively frequent, precluding the achievement of a diagnostic end-point in about 5 to 10% of tests. These adverse effects, mostly tachyarrhythmias and arterial hypotension, are usually minor and self limiting. However, severe life threatening complications, as well as death, also occur. By analysing Medline quoted literature up to March 1999, we found 35 original studies from a single institution with more than 100 patients, as well as 2 multicentre studies, concerning the feasibility and safety of dobutamine stress echocardiography. In a cumulative total of 26438 tests performed, 79 life-threatening complications (such as acute myocardial infarction, asystole, ventricular fibrillation, sustained ventricular tachycardia or severe symptomatic hypotension) have been reported, giving an incidence of 1 severe adverse reaction per every 335 examinations. In addition, 29 isolated case reports have been published describing life-threatening complications during dobutamine echocardiography. In case reports, 2 deaths have been described, both due to acute cardiac rupture in patients with recent inferior myocardial infarction. Severe adverse reactions during dobutamine echocardiography can be ischaemia independent, and are independent of operator experience and are unpredictable; some complications can be late occurring and long lasting. As a consequence, the procedure must be clearly indicated, written informed consent has to be obtained from the patient, an attending physician must be present during testing, and long term observation of outpatients is useful in order to manage late complications. In conclusion, while the safety of dobutamine stress echocardiography was reported to be outstanding in early reports, further experience presents a substantially more worrying picture. This must be taken into account by both physicians and patients when assessing the risk-benefit profile of the procedure. PMID- 10789824 TI - The tolerability of lamotrigine in children. AB - Lamotrigine is a novel anticonvulsant, which has proven to be effective both as add-on and monotherapy. 13 studies have demonstrated efficacy in 1096 children with a variety of seizure types. Tolerability information in these studies was collected in a standard fashion, where investigators reported all adverse events regardless of the perceived relationship to the test therapies. Generally, lamotrigine treatment in these clinical trials was generally given at higher initial doses and faster dose escalations than are currently recommended. Most adverse events associated with lamotrigine were mild to moderate in severity and did not result in discontinuation of treatment. Results from placebo-controlled, add-on trials showed that 85% of lamotrigine recipients experienced an adverse event compared with 83% of placebo recipients. Lamotrigine was associated with an increased risk of adverse events in the nervous system (dizziness, tremor, ataxia, and diplopia), gastrointestinal tract (nausea), and urinary tract (infection). The incidence of most adverse events was lower among lamotrigine recipients in monotherapy trials than in add-on trials, suggesting that concurrent anticonvulsant treatment or drug interactions can be confounding risk factors above that of lamotrigine treatment alone. Skin rash associated with hospitalisation and the discontinuation of study drug was reported more frequently by lamotrigine recipients than by placebo recipients and more frequently by children than by adults. The simultaneous use of valproic acid (sodium valproate) was associated with an increased incidence of rash. Lamotrigine, an effective broad spectrum anticonvulsant, is well tolerated in children. The qualitative features of adverse events that occur with lamotrigine treatment are similar for children and adults. The incidence of rash may be reduced with proper initial dosing and dose escalation. PMID- 10789825 TI - Comparative tolerability of sulphonylureas in diabetes mellitus. AB - The sulphonylurea drugs have been the mainstay of oral treatment for patients with diabetes mellitus since they were introduced. In general, they are well tolerated, with a low incidence of adverse effects, although there are some differences between the drugs in the incidence of hypoglycaemia. Over the years, the drugs causing the most problems with hypoglycaemia have been chlorpropamide and glibenclamide (glyburide), although this is a potential problem with all sulphonylureas because of their action on the pancreatic beta cell, stimulating insulin release. Other specific problems have been reported with chlorpropamide that occur only rarely, if at all, with other sulphonylureas. Hyponatraemia secondary to inappropriate antidiuretic hormone activity, and increased flushing following the ingestion of alcohol, have been well described. The progressive beta cell failure with time results in eventual loss of efficacy, as these agents depend on a functioning beta cell and are ineffective in the absence of insulin producing capacity. Differences in this secondary failure rate have been reported, with chlorpropamide and gliclazide having lower failure rates than glibenclamide or glipizide. The reasons for this are unclear, but the more abnormal pattern of insulin release produced by glibenclamide may be partly responsible and, indeed, may explain the increased risk of hypoglycaemia with this agent. Previously reported increased mortality associated with tolbutamide therapy has not been substantiated, and more recent data have shown no increased mortality from sulphonylurea treatment. Indeed, benefit from glycaemic control, regardless of the agent used--insulin or sulphonylurea--was reported by the United Kingdom Prospective Diabetes Study. Nevertheless, there is still ongoing controversy in view of the experimental evidence, mainly from animal studies, of potential adverse effects on the heart from sulphonylureas, but these are difficult to extrapolate into clinical situations. Most of these studies have been carried out with glibenclamide, which makes comparison of possible risk difficult. Other cardiovascular risk factors may be modified by gliclazide, which seems unique among the sulphonylureas in this respect. Its reported haemobiological and free radical scavenging activity probably resides in the azabicyclo-octyl ring structure in the side chain. Reduced progression or improvement in retinopathy has been reported in comparative trials with other sulphonylureas, and the effect is unrelated to improvements in glycaemia. There are differences between the sulphonylureas in some adverse effects, risk of hypoglycaemia, failure rates and actions on vascular risk factors. As a group of drugs, they are very well tolerated, but differences in overall tolerability can be identified. PMID- 10789826 TI - Drug-related problems in hospitalised patients. AB - Drug-related problems include medication errors (involving an error in the process of prescribing, dispensing, or administering a drug, whether there are adverse consequences or not) and adverse drug reactions (any response to a drug which is noxious and unintended, and which occurs at doses normally used in humans for prophylaxis, diagnosis or therapy of disease, or for the modification of physiological function). Furthermore, adverse drug events can be defined as an injury--whether or not causally-related to the use of a drug. Drug-related problems are relatively common in hospitalised patients and can result in patient morbidity and mortality, and increased costs. In order to get an overview of studies on drug-related problems in hospitalised patients, with specific attention to the incidence of drug-related problems and their costs, to the possibilities of prevention and to the effect of these interventions, we performed a literature search. Incidences of medication errors reported in studies vary widely. The range of reported incidences of adverse drug reactions is even wider. These wide ranges can be largely explained by the different study methods and definitions used. Problems related to drug therapy may be averted by preventive interventions. Several possibilities for prevention exist, especially for the prevention of medication errors. Prescribing, transcription and interpretation errors can be reduced by using computerised physician order entry. Together with the use of automated dispensing systems and bar-code technology, this will aid in the reduction of both dispensing and administration errors. Education of nursing staff involved in the process of drug distribution is another important measure for preventing medication errors. Finally, the introduction of systems for the early detection of adverse drug reactions may help to reduce problems related to drug therapy. Identifying risk factors that contribute to the development of adverse drug reactions, may aid in the prevention of these reactions. PMID- 10789828 TI - Temporal and spatial expression of Hoxa-2 during murine palatogenesis. AB - 1. Mice homozygous for a targeted mutation of the Hoxa-2 gene are born with a bilateral cleft of the secondary palate associated with multiple head and cranial anomalies and these animals die within 24 hr of birth (Gendron-Maguire et al., 1993; Rijli et al., 1993; Mallo and Gridley, 1996). We have determined the spatial and temporal expression of the Hoxa-2 homeobox protein in the developing mouse palate at embryonic stages E12, E13, E13.5, E14, E14.5, and E15. 2. Hoxa-2 is expressed in the mesenchyme and epithelial cells of the palate at E12, but is progressively restricted to the tips of the growing palatal shelves at E13. 3. By the E13.5 stage of development, Hoxa-2 protein was found to be expressed throughout the palatal shelf. These observations correlate with palatal shelf orientation and Hoxa-2 protein may play a direct or indirect role in guiding the palatal shelves vertically along side the tongue, starting with the tips of the palatal shelves at E13, followed by the entire palatal shelf at E13.5. 4. As development progresses to E14, the stage at which shelf elevation occurs, Hoxa-2 protein is downregulated in the palatal mesenchyme but remains in the medial edge epithelium. Expression of Hoxa-2 continues in the medial edge epithelium until the fusion of opposing palatal shelves. 5. By the E15 stage of development, Hoxa 2 is downregulated in the palate and expression is localized in the nasal and oral epithelia. 6. In an animal model of phenytoin-induced cleft palate, we report that Hoxa-2 mRNA and protein expression were significantly decreased, implicating a possible functional role of the Hoxa-2 gene in the development of phenytoin-induced cleft palate. 7. A recent report by Barrow and Capecchi (1999), has illustrated the importance of tongue posture during palatal shelf closure in Hoxa-2 mutant mice. This along with our new findings of the expression of the Hoxa-2 protein during palatogenesis has shed some light on the putative role of this gene in palate development. PMID- 10789827 TI - Intracellular signals coupled to muscarinic acetylcholine receptor activation in cerebral frontal cortex from hypoxic mice. AB - 1. The aim of the present work was to determine hypoxia-induced modifications in the cascade of intracellular events coupled to muscarinic acetylcholine receptor (mAChR) activation in brain. For this purpose, enzymatic activities were measured on normoxically incubated frontal cortical slices from mice exposed to hypobaric hypoxia for 72 hr. 2. We found that hypoxia induced alterations in several cerebral enzymatic basal activities: it increased nitric oxide synthase (NOS), but it decreased both membrane protein kinase C (PKC) and phospholipase C activities. 3. The mAChR agonist carbachol was found to increase phosphoinositide hydrolysis to greater values in hypoxic tissues than those found in normoxic conditions. Furthermore, a greater translocation of PKC in response to carbachol was observed in hypoxic tissues than in normoxic ones. 4. Besides, carbachol induced a drastic reduction of NOS activity in hypoxic brains, at concentrations that stimulated this enzyme activity in normoxic preparations. In the latter, inhibition is obtained only with high concentrations of the cholinergic muscarinic agonist. 5. These results pointed to a carbachol-mediated mAChR hyperactivity induced by hypoxic insult. 6. The possibility that these effects would account for a compensatory mechanism to diminish NOS hyperactivity, probably protecting for NO neurotoxic action in hypoxic brain, is also discussed. PMID- 10789823 TI - Cardiotoxicity of chemotherapeutic agents: incidence, treatment and prevention. AB - Cytostatic antibiotics of the anthracycline class are the best known of the chemotherapeutic agents that cause cardiotoxicity. Alkylating agents such as cyclophosphamide, ifosfamide, cisplatin, carmustine, busulfan, chlormethine and mitomycin have also been associated with cardiotoxicity. Other agents that may induce a cardiac event include paclitaxel, etoposide, teniposide, the vinca alkaloids, fluorouracil, cytarabine, amsacrine, cladribine, asparaginase, tretinoin and pentostatin. Cardiotoxicity is rare with some agents, but may occur in >20% of patients treated with doxorubicin, daunorubicin or fluorouracil. Cardiac events may include mild blood pressure changes, thrombosis, electrocardiographic changes, arrhythmias, myocarditis, pericarditis, myocardial infarction, cardiomyopathy, cardiac failure (left ventricular failure) and congestive heart failure. These may occur during or shortly after treatment, within days or weeks after treatment, or may not be apparent until months, and sometimes years, after completion of chemotherapy. A number of risk factors may predispose a patient to cardiotoxicity. These are: cumulative dose (anthracyclines, mitomycin); total dose administered during a day or a course (cyclophosphamide, ifosfamide, carmustine, fluorouracil, cytarabine); rate of administration (anthracyclines, fluorouracil); schedule of administration (anthracyclines); mediastinal radiation; age; female gender; concurrent administration of cardiotoxic agents; prior anthracycline chemotherapy; history of or pre-existing cardiovascular disorders; and electrolyte imbalances such as hypokalaemia and hypomagnesaemia. The potential for cardiotoxicity should be recognised before therapy is initiated. Patients should be screened for risk factors, and an attempt to modify them should be made. Monitoring for cardiac events and their treatment will usually depend on the signs and symptoms anticipated and exhibited. Patients may be asymptomatic, with the only manifestation being electrocardiographic changes. Continuous cardiac monitoring, baseline and regular electrocardiographic and echocardiographic studies, radionuclide angiography and measurement of serum electrolytes and cardiac enzymes may be considered in patients with risk factors or those with a history of cardiotoxicity. Treatment of most cardiac events induced by chemotherapy is symptomatic. Agents that can be used prophylactically are few, although dexrazoxane, a cardioprotective agent specific for anthracycline chemotherapy, has been approved by the US Food and Drug Administration. Cardiotoxicity can be prevented by screening and modifying risk factors, aggressively monitoring for signs and symptoms as chemotherapy is administered, and continuing follow-up after completion of a course or the entire treatment. Prompt measures such as discontinuation or modification of chemotherapy or use of appropriate drug therapy should be initiated on the basis of changes in monitoring parameters before the patient exhibits signs and symptoms of cardiotoxicity. PMID- 10789831 TI - Gene expression of two glutamate receptor subunits in response to repeated stress exposure in rat hippocampus. AB - 1. Glutamatergic mechanisms are thought to be involved in stress-induced alterations of brain function, especially in the hippocampus. We have hypothesized that repeated stress exposure may evoke changes of hippocampal glutamate receptors at the level of gene expression. 2. The study was designed to analyze the levels of mRNA coding for NMDAR1, the essential subunit of the N methyl-D-aspartate (NMDA) receptor subtype, and for GluR1, an AMPA glutamate receptor subunit, after repeated immobilization stress in rat hippocampus. Toward this aim, we applied a competitive RT-PCR technique which allowed precise and reliable quantification of the transcripts. 3. We found that repeated immobilization stress for 7 days significantly increased GluR1 mRNA levels, by 27% (P<0.01), as measured 24 hr after the last stress exposure. Levels of mRNA coding for NMDAR1 were slightly elevated, but the difference failed to be significant. 4. These results demonstrate selective changes in the gene expression of glutamate receptor subunits, which are likely to take part in the mechanisms leading to enhanced excitability and vulnerability of hippocampal neurons and to potential damage during repeated or chronic stress exposure. PMID- 10789830 TI - Functional coupling of G proteins to endothelin receptors is ligand and receptor subtype specific. AB - 1. The aims of the present study were (a) to determine the identity of the G proteins with which the endothelin receptor interacts and whether this interaction is subtype specific and (b) to determine whether agonist exposure can result in specific coupling between the endothelin receptor and G proteins. 2. Coupling between endothelin A (ET(A)) or endothelin B (ET(B)) receptors and G proteins was assessed in two fibroblast cell lines, each expressing one receptor subtype. Four ligands, ET-1, ET-3, SRTXb, and SRTXc, were used for receptor stimulation. The G protein alpha-subunit coupled to the receptor was identified by immunoprecipitation with an antibody against the endothelin receptor and immunoblotting with specific antibodies against different G protein alpha subunits. 3. Unstimulated ET(A) and ET(B) receptors (ET(A)R and ET(B)R, respectively) were barely coupled to Go(alpha). The unstimulated ET(A)R coimmunoprecipitated with Gi3alpha, whereas the unstimulated ETBR was much less strongly coupled to Gi3alpha. The coupling of ETBR to Gi1Gi2 alpha-subunits was much stronger than the coupling of ET(A)R to these alpha-subunits. Stimulation with the different ET agonists also resulted in differential coupling of G proteins to the receptor subtypes. All four ligands caused a strong increase in coupling of the ET(B)R to Gi3alpha, whereas coupling of the ET(A)R to this subunit was not affected by ET-1 and was even decreased by SRTXc. On the other hand, all four ligands caused a much greater increase in the coupling of ET(A)R to G(q)alpha/G11alpha than in the coupling of ET(B)R to these alpha-subunits. Ligand-induced coupling between the receptors and the Gi1 and Gi2 alpha-subunits is similar for the two receptor subtypes. The same was true for ligand-induced coupling of the receptors to Go(alpha), except that ET-3 increased the coupling of this alpha-subunit to ET(B)R and decreased the coupling to ET(A)R. Taken together, the results of this study show that coupling between ET receptors and G proteins is ligand and receptor subtype specific. 4. It remains to be established whether this diversity of receptor-G protein coupling is of relevance for the various endothelin signaling pathways and/or pathological states. PMID- 10789829 TI - Opioid and cannabinoid receptors share a common pool of GTP-binding proteins in cotransfected cells, but not in cells which endogenously coexpress the receptors. AB - 1. Opioid (mu, delta, kappa) and cannabinoid (CB1, CB2) receptors are coupled mainly to Gi/Go GTP-binding proteins. The goal of the present study was to determine whether different subtypes of opioid and cannabinoid receptors, when coexpressed in the same cell, share a common reservoir, or utilize different pools, of G proteins. 2. The stimulation of [35S]GTPgammaS binding by selective opioid and cannabinoid agonists was tested in transiently transfected COS-7 cells, as well as in neuroblastoma cell lines. In COS-7 cells, cotransfection of mu- and delta-opioid receptors led to stimulation of [35S]GTPgammaS binding by either mu-selective (DAMGO) or delta-selective (DPDPE) agonists. The combined effect of the two agonists was similar to the effect of either DAMGO or DPDPE alone, suggesting the activation of a common G-protein reservoir by the two receptor subtypes. 3. The same phenomenon was observed when COS-7 cells were cotransfected with CB1 cannabinoid receptors and either mu- or delta-opioid receptors. 4. On the other hand, in N18TG2 neuroblastoma cells, which endogenously coexpress CB1 and delta-opioid receptors, as well as in SK-N-SH neuroblastoma cells, which coexpress mu- and delta-opioid receptors, the combined effects of the various agonists (the selective cannabinoid DALN and the selective opioids DPDPE and DAMGO) were additive, implying the activation of different pools of G proteins by each receptor subtype. 5. These results suggest a fundamental difference between native and artificially transfected cells regarding the compartmentalization of receptors and GTP-binding proteins. PMID- 10789832 TI - Effects of A1 and A2 adenosine receptor antagonists on the induction and reversal of long-term potentiation in guinea pig hippocampal slices of CA1 neurons. AB - 1. Using simultaneous recordings of the field EPSP and the population spike in the CA1 neurons of guinea pig hippocampal slices, we confirmed that delivery of a high-frequency stimulation (tetanus: 100 pulses at 100 Hz) produced robust long term potentiation of synaptic efficacy (LTP) in two independent components, a synaptic component that increases field excitatory postsynaptic potentials (EPSPs) and a component that results in a larger population spike amplitude for a given EPSP size (E-S potentiation). 2. In the same cells, reversal of LTP (depotentiation; DP) in the field EPSP and in the E-S component is achieved by delivering low-frequency afferent stimulation (LFS: 1 Hz, 1000 pulses) 20 min after the tetanus. 3. When the tetanus or LFS was applied to CA1 inputs in the presence of an adenosine A1 receptor antagonist, 8-cyclopentyltheophylline (1 microM), the field EPSP was enhances in LTP and attenuated in DP, while the E-S relationship was not significantly affected in either LTP or DP. 4. When similar experiments were performed using an A2 receptor antagonist, CP-66713 (10 microM), the field EPSP was blocked in LTP but facilitated in DP, while E-S potentiation was enhanced during both LTP and DP. 5. The results show that endogenous adenosine, acting via A1 or A2 receptors, modulates both the synaptic and the E-S components of the induction and reversal of LTP. Based on the results, we discuss the key issue of the contribution of these receptors to the dynamics of neuronal plasticity modification in hippocampal CA1 neurons. PMID- 10789833 TI - Transient spinal cord ischemia in rat: the time course of spinal FOS protein expression and the effect of intraischemic hypothermia (27 degrees C). AB - 1. In the present study, we characterize the time course of spinal FOS protein expression after transient noninjurious (6-min) or injurious (12-min) spinal ischemia induced by inflation of a balloon catheter placed into the descending thoracic aorta. In addition, this work examined the effects of spinal hypothermia on FOS expression induced either by ischemia or by potassium-evoked depolarization (intrathecal KCl). 2. Short-lasting (6-min) spinal ischemia evoked a transient FOS protein expression. The peak expression was seen 2 hr after reperfusion in all laminar levels in lumbosacral segments. At 4 hr of reperfusion, more selective FOS expression in spinal interneurons localized in the central part of laminae V-VII was seen. At 24 hr no significant increase in FOS protein was detected. 3. After 12 min of ischemia and 2 hr of reflow, nonspecific FOS expression was seen in both white and gray matter, predominantly in nonneuronal elements. Intrathecal KCl-induced FOS expression in spinal neurons in the dorsal horn and in the intermediate zone. Spinal hypothermia (27 degrees C) significantly suppressed FOS expression after 6 or 12 min of ischemia but not after KCl-evoked depolarization. 4. Data from the present study show that an injurious (but not noninjurious) interval of spinal ischemia evokes spinal FOS protein expression in glial cells 2 hr after reflow. The lack of neuronal FOS expression corresponds with extensive neuronal degeneration seen in this region 24 hr after reflow. Noninjurious (6-min) ischemia induced a transient, but typically neuronal FOS expression. The significant blocking effect of hypothermia (27 degrees C) on the FOS induction after ischemia but not after potassium-evoked depolarization also suggests that simple neuronal depolarization is a key trigger in FOS induction. PMID- 10789834 TI - Time course of brain neuronal degeneration and heat shock protein (72) expression following neck tourniquet-induced cerebral ischemia in the rat. AB - 1. The present study was designed to examine the regional expression of HSP72/73 protein after a 7.5-min period of cerebral ischemia and to compare the distribution of HSP neurons with the localization of irreversible neuronal degeneration as analyzed by silver impregnation technique. 2. During 6-24 hr after cerebral ischemia clear-cut neuronal argyrophilia developed in several brain regions including the hippocampal hilus, nucleus reticularis thalami, and colliculi inferiores. With the exception of the hippocampal hilus, the structures which showed silver impregnability were HSP72 negative at 6-24 hr. 3. Despite the clear HSP72 expression seen in hippocampal CA1 neurons, a significant loss of these neurons was seen at 7 days after ischemia. 4. These data show that in some structures the presence of HSP72 is indicative of higher resistance of these neurons to ischemia-induced degeneration, however, the process of delayed neuronal degeneration appears to be independent of the accelerated synthesis of HSP72 seen during the early period of reflow. PMID- 10789835 TI - Critical temporal modulation of neuronal programmed cell injury. AB - 1. As a free radical, nitric oxide (NO) may be toxic to neurons through mechanisms that directly involve DNA damage. Lubeluzole, a novel benzothiazole compound, has recently been demonstrated to be neuroprotective through the signal transduction pathways of NO. We therefore examined whether neuroprotection by lubeluzole was dependent upon the molecular pathways of programmed cell death (PCD). 2. In primary hippocampal neurons, evidence of PCD was determined by hematoxylin and eosin (H&E) stain, transmission electron microscopy, and annexin V binding. NO administration with the NO generators sodium nitroprusside (300 microM) or SIN-1 (300 microM) directly induced PCD. 3. Neurons positive for PCD increased from 22+/-3% (untreated) to 72+/-3% (NO) over a 24-hr period. Coadministration of NO and lubeluzole (750 nM), a neuroprotective concentration, actively decreased PCD expression on H&E stain from 72+/-3% (NO only) to 25+/-3% (NO and lubeluzole). Significant reduction in DNA fragmentation by lubeluzole also was evident on electron microscopy. Application of lubeluzole in concentrations that were not neuroprotective or administration of the biologically inactive R-isomer did not significantly alter NO-induced PCD, suggesting that neuroprotection by lubeluzole was intimately linked to the modulation of PCD. Lubeluzole also was able to prevent the initial stages of cellular membrane inversion labeled with annexin-V binding, an early and sensitive indicator of PCD. Interestingly, the critical period for lubeluzole to reverse PCD induction appeared to be within the first 4 hr following NO exposure. 4. Further investigation into the neuroprotective pathways that alter PCD may provide greater insight into the molecular mechanisms that ultimately determine neuronal injury. PMID- 10789836 TI - Role of synaptophysin in exocytotic release of dopamine from Xenopus oocytes injected with rat brain mRNA. AB - 1. The role of synaptophysin in the exocytotic release of dopamine (DA) was examined in Xenopus laevis oocytes injected with rat brain mRNA. 2. The mRNA injected oocytes showed DA uptake which depended on the incubation time and external DA concentrations. 3. Stimulation with KCl (10-50 mM) of mRNA-injected oocytes preloaded with DA evoked external Ca2+ -dependent release of DA. The noninjected and water-injected oocytes did not produce uptake of DA and stimulation-evoked release of DA. 4. The high-KCl (50 mM)-stimulated release of DA decreased in the oocytes injected with rat brain mRNA together with antibody to synaptophysin. 5. Immunoblot analysis demonstrated that synaptophysin was expressed in the brain mRNA-injected oocytes but not in the noninjected and water injected oocytes. 6. Thus, uptake and release machinery similar to native dopaminergic nerve terminals was expressed in Xenopus oocytes by injecting mRNA extracted from the rat brain, and synaptophysin may play a role in the exocytotic release of DA. PMID- 10789838 TI - The importance of exposure estimation in the assessment of skin sensitization risk. AB - The development of new ingredients and products for the consumer market requires a thorough assessment of their potential for skin sensitization and the possible clinical manifestation of allergic contact dermatitis. The process by which low molecular weight chemicals induce and elicit skin sensitization reactions is complex and dependent on many factors relevant to the ability of the chemical to penetrate the skin, react with protein, and trigger the cell-mediated immune response. These major factors include inherent potency, chemical dose, duration and frequency of exposure, vehicle or product matrix, and occlusion. The fact that a chemical is a contact allergen does not mean that it cannot be formulated into a consumer product at levels well tolerated by most individuals. Many common ingredients (e.g., fragrances, preservatives) are known skin allergens. However, all allergens show dose-response and threshold characteristics. Therefore, one should be able to incorporate these chemicals into products at levels that produce acceptably low incidences of skin sensitization under foreseeable conditions of exposure. The critical exposure determinant for evaluating skin sensitization risk is dose per unit area of skin exposed. Use of this parameter allows for comparative assessments from different types of skin sensitization tests (including cross-species comparisons), and, at least for known potent allergens, there is remarkable similarity in threshold dose/unit area determinations across species. The dose/unit area calculation enables a judgment of the sensitization risk for different product types. This is illustrated using the chemical preservative methylchloroisothiazolinone/methylisothiazolinone (MCI/MI) as a case study. PMID- 10789837 TI - Engineering viral promoters for gene transfer to human neuroblasts. AB - 1. The strength and activity of several viral promoters in human neuroblasts were evaluated in vitro. 2. Several luciferase reporter gene constructs under the control of different viral promoters (HIV-1 LTR, HTLV-I LTR, MMTV LTR, RSV LTR, CMV, SV40), in the presence or in the absence of the viral SV40 enhancer, were transfected into two well-established human neural cell lines, including one derived from human embryonic olfactory cells (B4) and one derived from an adrenal neuroblastoma (SH-SY-5Y). The epithelial cell line HeLa was used as a control. 3. The enzymatic activity of luciferase was evaluated after normalization with an internal control. The results indicated that in the context of the reporter gene constructs, the CMV promoter alone was, overall, the most active in any tested cell line. However, addition of the SV40 enhancer to the CMV promoter abolished luciferase activity in SH-SY-5Y cells while significantly increasing luciferase expression in the CNS derived B4 fetal neuroblasts. 4. The results suggest that gene therapeutic vectors aimed to promote enzymatic activity through gene transfer into undifferentiated human neural cells are feasible. However, since differences in promoter activity in neuroectodermal-derived cells are very relevant, gene construct variants should be considered to optimize the system. PMID- 10789839 TI - Placebo-controlled evaluation of the irritant potential of tacalcitol (1a,24 dihydroxyvitamin D3) in healthy volunteers. AB - In the treatment of psoriasis with topical vitamin D3 analogues, lesional and perilesional irritation is the main side-effect. The aim of this study was to investigate whether local side-effects generated by tacalcitol, a vitamin D3 analogue, show concentration dependence. 3 different concentrations of tacalcitol (0.4; 4; 40 microg/g ointment) and the vehicle were applied on normal skin of the back of 25 healthy volunteers under occlusive conditions for 5 days. Assessment of erythema, infiltration and scaling as well as measurement of transepidermal water loss (TEWL) was performed on days 1 to 5. On day 5, additional skin barrier tests (DMSO test, alkali resistance test) were performed. Erythema and slight infiltration, but no scaling, were observed in a number of subjects without significant differences. TEWL also did not show significant differences for the test formulations, though there was a tendency towards lower values in the untreated areas. In the skin barrier tests, a tendency towards higher alkali resistance in the test areas treated with 40 microg tacalcitol/g ointment was detected. Thus, under occlusive conditions, the irritant potential of tacalcitol is very low. There is no convincing evidence of concentration dependence in irritation generated by tacalcitol when applied under occlusive conditions. PMID- 10789840 TI - Chronic irritant contact dermatitis: recovery time in man. AB - Chronic irritant contact dermatitis (ICD) is a common skin disease, especially in the workplace, but determining the recovery time of chronic ICD is not easy. To measure the recovery time of chronic ICD, we examined the skin reactivity to a model surfactant, sodium lauryl sulfate (SLS), on previous chronic ICD and normal sites by visual grade and non-invasive instruments. Chronic ICD was induced on the forearms of 10 volunteers (aged 23 to 43 years) by occluded application of 1% SLS for 30 min on 5 consecutive days each week for 3 weeks. Previous ICD and normal sites were provoked by the occluded application of 7.5% SLS for 30 min daily on 4 consecutive days, 2, 5 and 10 weeks after induction. Skin reactivity was assessed daily by awarding visual erythema scores, visual scale scores and measuring transepidermal water loss, skin color reflectance, and electrical capacitance. Skin reactivity of previous chronic ICD sites to SLS showed hyperreactivity compared to normal sites even after the 10th week post-induction. PMID- 10789841 TI - Experimental study on skin sensitization potencies and cross-reactivities of hair dye-related chemicals in guinea pigs. AB - In screening patch testing of hairdressers with occupational contact dermatitis, multiple positive reactions to hair dye-related chemicals, such as p phenylenediamine (PPD), p-toluenediamine x 2HCl (PTD) and p-aminophenol (PAP), a fabric dye p-aminoazobenzene (PAB), and a tar dye Sudan III, were frequently encountered. To investigate individual skin sensitization potency and the cross reactivities among above chemicals, a guinea pig maximization test with the above 5 chemicals was performed. In each group, 6 animals were induced with one of the chemicals at 0.1% concentration by intradermal injection and at 1.0% by topical application. The animals were challenged with all 5 chemicals in concentrations of dilution by 10 from 0.1% to 0.001%. Under the conditions of 0.1% challenges, similar sensitization potencies were observed in PPD (6/6), PTD (6/6), PAP (5/6) and PAB (6/6) groups, but no positive reactions were elicited in the Sudan III group. The cross-reactivities to PPD were confirmed in the animals challenged with PTD (6/6), PAP (6/6), PAB (6/6) and Sudan III (3/6). In the PTD-induced group, positive responses to cross-challenges were elicited by PPD (5/6), PAP (3/6), PAB (5/6) and Sudan III (1/6). The cross-reactivities to PAP were observed only with PPD (2/5) and PAB (5/5). PAB-induced animals responded only to PPD (1/6). The results indicate that all these chemicals except Sudan III are strong sensitizers. Their cross-reactivities are different in sensitized conditions, respectively. The cross-reactivities to PPD were higher than those to PTD, PAP and PAB. PMID- 10789842 TI - Contact allergy to Psorigon. PMID- 10789843 TI - Oral tolerance of nickel in patients with dyshidrosis. PMID- 10789844 TI - Occupational contact dermatitis in fishermen of Cumbuco beach, Ceara, northeast Brazil. PMID- 10789846 TI - Eruption caused by a deep-sea cnidarian. PMID- 10789845 TI - Delayed allergic skin reactions to subcutaneous heparins. Tolerance of 2 recombinant hirudins. PMID- 10789847 TI - Protein contact dermatitis: aimed testing with foods. PMID- 10789848 TI - Allergic contact dermatitis from dibucaine hydrochloride. PMID- 10789849 TI - Reaction to cocamidopropyl hydroxysultaine, an amphoteric surfactant and conditioner. PMID- 10789850 TI - Fixed drug eruption induced by tosufloxacin tosilate. PMID- 10789851 TI - Iodine allergy induced by consumption of iodine-containing food. PMID- 10789852 TI - Maculopapular eruption from sertraline with positive patch tests. PMID- 10789853 TI - Allergy to rubber additives in orthopedic braces. PMID- 10789854 TI - False-negative patch test reactions due to a lower concentration of patch test substance than declared. PMID- 10789855 TI - Eyelid dermatitis: an evaluation of 232 patch test patients over 5 years. PMID- 10789856 TI - Very delayed reactions to beta-lactam antibiotics. PMID- 10789857 TI - Eczematous plaques related to unfractionated and low-molecular-weight heparins: cross-reaction with danaparoid but not with desirudin. PMID- 10789858 TI - Eosinophilic cationic proteins in the peripheral blood of patients with contact allergy. PMID- 10789860 TI - Results of patch testing with hydrocortisone butyrate in different vehicles. PMID- 10789859 TI - Type I and type IV immune responses to glyceryl thioglycolate. PMID- 10789861 TI - Allergic contact dermatitis due to Icelandic poppy (Papaver nudicaule). PMID- 10789862 TI - Gold sensitivity in Israel--consecutive patch test results. PMID- 10789863 TI - Polyhexamethylenebiguanide: a relevant contact allergen? PMID- 10789864 TI - Localized aquagenic urticaria. PMID- 10789865 TI - Recurrent rejection of a spinal cord stimulation system. PMID- 10789866 TI - Allergic contact dermatitis due to clobetasone butyrate. PMID- 10789867 TI - Epoxy-by-proxy dermatitis. PMID- 10789868 TI - Allergic contact dermatitis from benzalkonium chloride in an antifungal solution. PMID- 10789869 TI - Dihydrocodeine: a drug allergy diagnosed by patch testing. PMID- 10789870 TI - An unusual cause of post-auricular dermatitis. PMID- 10789871 TI - Combined contact allergy to tea tree oil and lavender oil complicating chronic vulvovaginitis. PMID- 10789872 TI - Possible allergy to complex titanium salt. PMID- 10789873 TI - Contact dermatitis from tosylamide/formaldehyde resin with photosensitivity. PMID- 10789874 TI - Flare-up reaction of pseudoephedrine baboon syndrome after positive patch test. PMID- 10789875 TI - Predicting haloperidol occupancy of central dopamine D2 receptors from plasma levels. AB - Positron emission tomography (PET) is increasingly being used to study dopamine receptor occupancy and the clinical effects of antipsychotic medication. Dopamine D2 receptor occupancy has been shown to predict several clinical effects of antipsychotic medication including therapeutic response, motor and endocrine side effects. Plasma levels may be used as a surrogate marker for central occupancy if the relationship between these two measures may be accurately described. This study was designed to test the capacity of a previously derived relationship equation (%D2 occupancy=plasma level/ED50+plasma level, where ED50= 0.40 ng/ml) to predict striatal D2 occupancy from plasma level. Twenty-one patients receiving treatment with low dose haloperidol underwent a 11C-raclopride PET scan to measure D2 occupancy. The D2 occupancy levels were accurately predicted by use of the previously generated equation with only a small degree of error (3.89% CI 0.45-7.33). Predicted and measured D2 occupancy values correlated closely (Pearson's r=0.864, P=0.003). The study indicates that reliable prediction of D2 occupancy from plasma levels is possible. This provides a potentially useful surrogate measure of D2 occupancy for research and possibly clinical practice, as the routine use of PET to measure occupancy levels is not feasible. PMID- 10789876 TI - Attenuation of brain high frequency electrocortical response after thiopental in early stages of Alzheimer's dementia. AB - RATIONALE: Pathological brain regions generate proportionately less high frequency (beta) activity than non-pathological regions, a phenomenon accentuated by barbiturate administration. OBJECTIVES: Previously, we reported a loss of high frequency brain electrical response to thiopental in dementia of the Alzheimer's type (DAT). The current study examines whether this phenomenon may be detected in early stages of the illness. METHODS: Using quantitative electroencephalography, we examined power in the 20-28 Hz band in patients with early DAT (n=7, age 71.0+/-3.2 years, Folstein Mini Mental State Score, MMSE 26.2+/-0.8), normal controls (n=8, age 74.3+/-3.2 years, MMSE 29.0+/-0.3) and subjects with moderately severe DAT (n=6, age 76.6+/-3.0 years, MMSE=12.5+/-3.7) at baseline and following an intravenous bolus of thiopental (0.5 mg/kg). RESULTS: No significant group differences in beta power were detectable at baseline. In response to thiopental, early DAT subjects compared to controls showed a significantly smaller beta power response in the frontal region at 1-3 min postinjection. Losses were smaller than those of subjects with moderately severe DAT and demonstrated a non-linear correlation with decreases in cognitive function as assessed by the MMSE score (r2=0.75). CONCLUSION: In early stages of DAT, a barbiturate challenge may unmask abnormalities in brain electrical activity not seen at baseline. Such changes may reflect underlying cortical deafferentation. PMID- 10789877 TI - Effects of lithium on platelet membrane phosphoinositides in bipolar disorder patients: a pilot study. AB - RATIONALE: In vitro and in vivo animal studies suggest that the intracellular phosphatidylinositol (PI) pathway is an important target for the effects of lithium. OBJECTIVES: We conducted a preliminary study to examine the in vivo effects of lithium treatment on platelet membrane phosphoinositides in bipolar disorder subjects, in an attempt to examine further the hypothesis that lithium has significant in vivo effects on the PI pathway in these patients. METHODS: We quantitated PI, phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol 4,5-bisphosphate (PIP2) in platelet membranes of seven subjects (five male, two female; mean age= 27.9+/-5.7 years), initially while they were unmedicated, and a second time after at least 21 days of lithium treatment (mean+/-SD=28.7+/-7.1 days). RESULTS: The mean+/-SD values for PI were 5.63+/-2.25% and 5.21+/-1.06%; for PIP 0.68+/-0.20% and 0.55+/-0.11%; and for PIP2 0.60+/-0.21% and 0.38+/ 0.15%, before and after lithium treatment, respectively. The decrease in PIP2 values after lithium treatment was statistically significant (Wilcoxon signed ranks test, Z=-2.37, P=0.02). CONCLUSION: This longitudinal study suggests that therapeutic doses of lithium significantly decrease platelet membrane PIP2 levels in vivo in bipolar disorder subjects, which may be related to lithium's mechanism of action in bipolar disorder. PMID- 10789878 TI - Inhibitory effects of clozapine and other antipsychotic drugs on noradrenaline transporter in cultured bovine adrenal medullary cells. AB - The effects of clozapine and other antipsychotic drugs on noradrenaline (NA) transport were examined in cultured bovine adrenal medullary cells and in transfected Xenopus laevis oocytes expressing the bovine NA transporter. Incubation of adrenal medullary cells with clozapine (30-1000 ng/ml) inhibited desipramine (DMI)-sensitive uptake of [3H]NA in a concentration-dependent manner (IC50=110 ng/ml or 336 nM). Other antipsychotic drugs such as haloperidol, chlorpromazine, and risperidone also decreased [3H]NA uptake (IC50= 144, 220, and 210 ng/ml or 383, 690, and 512 nM, respectively). Eadie-Hofstee analysis showed that clozapine reduced V(max) of uptake of [3H]NA and increased K(m). Furthermore, clozapine inhibited specific binding of [3H]DMI to plasma membranes isolated from bovine adrenal medulla (IC50=48 ng/ml or 146 nM). Scatchard plot analysis of [3H]DMI binding revealed that clozapine decreased both B(max) and K(d). Other antipsychotic drugs, including haloperidol, chlorpromazine, and risperidone, also reduced [3H]DMI binding to the membranes. In transfected Xenopus oocytes expressing the bovine NA transporter, clozapine inhibited [3H]NA uptake in a concentration-dependent manner similar to that observed in adrenal medullary cells. These results suggest that clozapine and haloperidol directly inhibit transport of NA by acting on the site of an NA transporter that influences both substrate transport and binding of tricyclic antidepressants. PMID- 10789879 TI - Conditioned effects of environmental stimuli paired with smoked cocaine in humans. AB - RATIONALE: Clinical data suggest that stimuli paired with cocaine use acquire emergent stimulus effects, such as the ability to elicit cocaine craving. OBJECTIVES: The purpose of this study was to determine the conditioned effects of neutral stimuli paired with cocaine smoking. METHODS: Eight experienced adult cocaine smokers participated in 22 experimental sessions while residing on a Clinical Research Center. One set of cues (CS-) was paired with placebo smoked cocaine and one set of cues (CS+) was paired with 25 mg smoked cocaine. RESULTS: After 18 training trials, the effects of cocaine on heart rate and ratings of "anxious" were greater, and skin temperature and ratings of "tired" were smaller when compared to the effects of cocaine after the first training trial. When instructed to select a cue to experience after training, seven of eight participants selected the CS+, while only three of the participants selected the CS+ prior to training, i.e., the CS+ functioned as a conditioned reinforcer. Presentation of the CS+ alone without cocaine during extinction trials increased HR, SP, and ratings of "anxious" "tired", and "I want cocaine" and decreased skin temperature. These changes elicited by presentation of the CS+ decreased over the course of the extinction sessions. CONCLUSIONS: The present results indicate that classical conditioning is one mechanism by which stimuli paired with cocaine acquire emergent stimulus effects. PMID- 10789881 TI - Antidepressant-like effect of 7-nitroindazole in the forced swimming test in rats. AB - RATIONALE: There is some strong evidence about the role of nitric oxide (NO) as an intercellular messenger in central physiological mechanisms. NO is synthesized from L-arginine by nitric oxide synthase (NOS), as a response to activation of N methyl-D-aspartate (NMDA) receptors by excitatory amino acids. NMDA receptor antagonists also produce antidepressant-like actions in preclinical models. OBJECTIVE: In the present study, the involvement of NO in the mechanism of depression was investigated. 7-Nitroindazole (7-NI) (15, 30, 60, 90 mg/kg IP), a selective inhibitor of neuronal NOS was examined. METHODS: The Porsolt forced swimming test (FST) has been used as a test for screening new antidepressant agents. RESULTS: 7-NI dose-dependently decreased the immobility time in FST, but produced no significant change in locomotor activity in naive rats. Neither L arginine, nor D-arginine (100 mg/kg) affected the immobility time in the FST or revealed any effect on locomotion. L-Arginine but not D-arginine, given 10 min before 7-NI, reversed the 7-NI-induced effect on immobility time. CONCLUSIONS: Our findings suggest that NO might be an important modulator of depression in rats. PMID- 10789880 TI - Diazepam-like effects of a fish protein hydrolysate (Gabolysat PC60) on stress responsiveness of the rat pituitary-adrenal system and sympathoadrenal activity. AB - RATIONALE: Gabolysat PC60 is a fish protein hydrolysate with anxiolytic properties commonly used as a nutritional supplement. OBJECTIVE: The diazepam like effects of PC60 on stress responsiveness of the rat pituitary-adrenal system and on sympathoadrenal activity were studied. METHODS: The activity of the pituitary-adrenal axis, measured by plasma levels of adrenocorticotropic hormone (ACTH) and corticosterone (B) of the sympathoadrenal complex, measured by circulating levels of noradrenaline (NA) and adrenaline (A), and the gamma aminobutyric acid (GABA) content in the hippocampus and the hypothalamus were investigated in male rats which received daily, by an intragastric feeding tube, for 5 days running either diazepam (1 mg/kg) or PC60 (300 or 1,200 mg/kg). Controls received only solvent (carboxymethylcellulose 1%). Six hours after the last force-feeding, the rats were subjected to 3 min ether inhalation or 30 min restraint and killed by decapitation 30 min after ether stress or at the end of restraint. RESULTS: Baseline plasma levels of ACTH, B, NA and A were not affected by either diazepam or PC60. Both ether- and restraint-induced release of ACTH, but not B, were similarly and drastically reduced by diazepam and PC60 (1,200 mg/kg). Both diazepam and PC60 (1,200 mg/kg) deleted restraint-induced NA and A increases. Both treatments also reduced the ether-induced rise of A. Basal levels of GABA were significantly increased in both the hippocampus and the hypothalamus in PC60-treated rats and only in the hippocampus in diazepam-treated ones. In controls, ether inhalation as well as restraint increased GABA content of these two brain structures. In contrast, such stress procedures performed in PC60 treated rats reduced GABA content slightly in the hippocampus but significantly in the hypothalamus. In diazepam-treated rats, GABA content of the hypothalamus was unaffected by stresses but that of the hippocampus was slightly decreased. CONCLUSIONS: Present data suggest diazepam-like effects of PC60 on stress responsiveness of the rat pituitary adrenal axis and the sympathoadrenal activity as well as GABA content of the hippocampus and the hypothalamus under resting and stress conditions. These effects of PC60 agree with anxiolytic properties of this nutritional supplement, previously reported in both rats and humans. PMID- 10789882 TI - Z-IQNP: a potential radioligand for SPECT imaging of muscarinic acetylcholine receptors in Alzheimer's disease. AB - RATIONALE: The density of the M2 subtype of muscarinic acetylcholine receptors (mAChR) has been shown to be reduced in the brain of patients with Alzheimer's disease (AD). It is therefore of interest to develop a brain imaging method for diagnostic purposes. Z-(R,R)-1-azabicyclo[2.2.2]oct-3-yl alpha-hydroxy-alpha-(1 iodo1-propen-3-yl)-alpha-phenylacetat e (Z-IQNP) is a muscarinic antagonist with high affinity for the M2 subtype. OBJECTIVE: The pharmacological characteristics and topographic distribution of radiolabelled Z-IQNP as a radioligand for the M2 mAChR subtype were examined in vitro and in vivo. METHODS: Z-IQNP was labelled with 1251 and 123I. Autoradiography was performed on whole-hemisphere cryosections from human post mortem brains. SPECT was performed in a cynomolgus monkey. RESULTS: Autoradiography showed binding of [125I]Z-IQNP in all brain regions, which was inhibited by the non-selective muscarinic antagonist scopolamine. The addition of BIBN 99, a compound with high affinity for the M2 subtype, inhibited [125I]Z-IQNP binding particularly in the cerebellum, which has a high density of the M2 subtype. SPECT demonstrated high uptake of [123I]Z-IQNP in all brain regions. The binding was markedly reduced in all brain regions after pretreatment with the non-selective muscarinic antagonist dexetimide and also the M1 antagonist biperiden. Dexetimide markedly inhibited [123I]Z-IQNP binding in the cerebellum, which is consistent with a high density of M2-receptors in this region. The sigma receptor binding compound DuP 734 had no effect on Z-IQNP binding either in vitro or in vivo. CONCLUSIONS: This study indicates that radiolabelled Z-IQNP has high specificity for mAChR with higher affinity for the M2 than the M1 subtype and negligible affinity for sigma recognition sites both in vitro and in vivo. [123I]Z-IQNP should be useful for future SPECT studies in AD for examination of the density of M2 receptors particularly in the cerebellum. PMID- 10789883 TI - Acute exposure to saccharin reduces morphine analgesia in the the rat: evidence for involvement of N-methyl-D-aspartate and peripheral opioid receptors. AB - RATIONALE: Pairings of a sweet taste and injection of morphine result in a learned avoidance of that taste and learned analgesic tolerance. This avoidance is mediated by the drug's peripheral effect, while learned tolerance involves activation of N-methyl-D-aspartate (NMDA) receptors. Exposure to a sweet taste also reduces morphine analgesia. We studied whether this taste-mediated reduction was reversed by an NMDA or peripheral opioid receptor antagonist. OBJECTIVES: To determine whether an intraoral infusion of saccharin would modulate morphine analgesia in rats, and to study the contribution of NMDA as well as peripheral opioid receptors to this modulation. METHODS: Six experiments used the rat's tail flick response to study the effect of an intraoral infusion of a sodium saccharin solution on morphine analgesia, and the effects of the quaternary opioid receptor antagonist methylnaltrexone as well as the noncompetitive NMDA receptor antagonist MK-801 on this modulation of analgesia. RESULTS: An intraoral infusion of saccharin reduced the analgesic effects of an intraperitoneal (i.p.) injection of morphine across a range of doses (experiment la), which was not attributable to an influence on tail-skin temperature (experiment 1b). This reduction was mediated by opioid receptors in the periphery and activation of NMDA receptors because morphine analgesia was reinstated by an i.p. injection of either methylnaltrexone (experiment 2a) or MK-801 (experiment 3a), which was not due to the effect of methylnaltrexone (experiment 2b) or MK-801 (experiment 3b) on morphine analgesia in the absence of saccharin. CONCLUSIONS: These results document evidence for an antagonism of morphine analgesia by actions of the drug at peripheral opioid receptors and excitatory amino-acid activity at NMDA receptors. They are discussed with reference to the aversive motivational effects of peripheral opioid receptors and pain facilitatory circuits. PMID- 10789884 TI - Glucose administration, heart rate and cognitive performance: effects of increasing mental effort. AB - RATIONALE: It is known that glucose administration is capable of improving performance on tests of declarative verbal memory and non-mnemonic tasks requiring high "mental effort". At the same time, cognitively demanding tasks are associated with elevated heart rate, a response that could feasibly be part of a physiological mechanism serving to increase the delivery of glucose to active brain substrates. OBJECTIVE: The present placebo-controlled, double-blind, balanced, crossover study examined the interaction between glucose administration, cognitive performance and heart rate during three tasks of differing mental demand and somatically-matched control tasks. METHODS: The effects of a glucose drink on participants' performance on two serial subtraction tasks (Serial Threes and Serial Sevens) and a Word Retrieval (Verbal Fluency) task were assessed. Heart rates were monitored throughout the experiment, and participants rated each task in terms of its perceived mental demand. RESULTS: Serial Sevens was rated as the most mentally demanding task, followed by Word Retrieval, then Serial Threes. Glucose consumption significantly improved performance on Serial Sevens, with a trend for improved performance on Word Retrieval. Both Serial Sevens and Serial Threes were associated with significant heart rate elevation above that seen in somatically matched control tasks (ruling out the possibility that accelerated heart rate was due to peripheral mechanisms alone). Unexpectedly, participants in the glucose condition had higher heart rates during cognitive processing. Additionally, individuals whose baseline heart rates were below the median performed better on Serial Threes and Serial Sevens. CONCLUSION: We suggest that supplemental glucose preferentially targets tasks with a relatively high cognitive load, which itself (through unknown mechanisms) mobilises physiological reserves as part of a natural response to such tasks. Furthermore, baseline heart rate and responses to cognitive demand and glucose administration may represent important physiological individual differences. PMID- 10789885 TI - Comparison of the antidepressants reboxetine, fluvoxamine and amitriptyline upon spontaneous pupillary fluctuations in healthy human volunteers. AB - RATIONALE: Spontaneous fluctuations in the size of the pupil in darkness are a recognised index of "sleepiness". OBJECTIVE: To evaluate the effects of single oral doses of three antidepressants: reboxetine (4 mg), a selective noradrenaline reuptake inhibitor, fluvoxamine (100 mg), a selective serotonin reuptake inhibitor, and amitriptyline (100 mg), a tricyclic antidepressant of known sedative property, upon spontaneous pupillary fluctuations in healthy male volunteers (n=16). METHODS: Using the recently developed pupillographic sleepiness test (PST), resting pupil diameter was recorded and two measures of pupillary fluctuations were obtained: total power obtained from a fast Fourier transform and spectral analysis, and the pupillary unrest index (PUI), a cumulative measure of changes in pupil size. Subjects also rated themselves on a battery of visual analogue scales for "alertness", "anxiety" and "contentedness". RESULTS: Resting pupil diameter was enhanced by reboxetine, but remained unaffected by the other two antidepressants. Amitriptyline, consistent with its sedative property, increased the total power of pupillary fluctuations and showed a tendency to increase PUI. These pupillary effects of amitriptyline were paralleled by reduced scores on the "alertness", "contentedness" and "anxiety" self ratings. Neither fluvoxamine nor reboxetine affected pupillary fatigue waves or subjective ratings of "alertness". Reboxetine caused a small reduction in subjectively rated "anxiety". CONCLUSIONS: The mydriatic effect of reboxetine may be due to noradrenaline reuptake blockade in the iris and/or in the central nervous system. The enhancement of pupillary fatigue waves by the sedative antidepressant amitriptyline, but not by the non-sedative antidepressants fluvoxamine and reboxetine, indicates that the PST is a suitable quantitative objective test for the detection of drug-induced changes in the level of arousal. PMID- 10789886 TI - Comparison of either norepinephrine-uptake inhibitors or phentermine combined with serotonergic agents on food intake in rats. AB - RATIONALE: We have shown previously that the anorectic effects of the catecholamine-releasing agent phentermine (PHEN) and the serotonin (5-HT) releasing agent dexfenfluramine (DFEN) are greater than additive in rats. In the present study, we examined whether the norepinephrine-uptake inhibitors desmethylimipramine (DMI) and thionisoxetine (TNIX) have additive effects with either DFEN or with the 5-HT-uptake inhibitor fluoxetine (FLX). We also examined whether PHEN interacts with a postsynaptically acting 5-HT agonist. METHODS: Undeprived rats were trained to eat a daily sweet-milk dessert and on test days were systemically administered single or combination drugs and the intakes recorded. RESULTS: Both DMI and TNIX produced dose-related suppressions of food intake. However, by isobolographic analysis, they did not enhance the anorectic actions of either DFEN or FLX. In contrast, confirming and extending our previous work, PHEN had a greater potentiating effect on the anorectic actions of DFEN and FLX than TNIX. Further, the anorectic action of the 5-HT2c receptor agonist TFMPP was enhanced by PHEN. CONCLUSIONS: These and other data are consistent with the idea that 5-HT agents may work "upstream" of critical catecholaminergic synapses in the production of anorexia, and explain the diminished efficacy of norepinephrine-uptake inhibitors relative to PHEN. The implications for clinically useful anorectic agents are discussed briefly. PMID- 10789887 TI - Modulation of the ethanol-like discriminative stimulus effects of diazepam and phencyclidine by L-type voltage-gated calcium-channel ligands in rats. AB - RATIONALE: Administration of voltage-gated calcium-channel (VGCC) modulators with ethanol can result in enhancement or attenuation of some behavioral effects of ethanol, including its discriminative stimulus effects. OBJECTIVES: The present study used a drug-discrimination paradigm to characterize modulation of the ethanol-like discriminative stimulus effects of a gamma-amino-butyric acid (GABA)A and N-methyl-D-aspartate (NMDA) ligand by administration of VGCC ligands. METHODS: Two groups of adult male Long-Evans rats were trained to discriminate either 1.0 g/kg ethanol (n=8) or 2.0 g/kg ethanol (n=9) from water under a fixed ratio (FR) 20 schedule of food presentation. Following training, ethanol substitution tests were conducted with cumulative doses of the GABA(A)-positive modulator diazepam (0.3-10 mg/kg, i.p.) (DZP) and the uncompetitive NMDA antagonist phencyclidine (0.3-5.6 mg/kg, i.p.) (PCP). Next, a single dose of the VGCC antagonist nimodipine, nifedipine, isradipine, or the VGCC agonist (-)-BAY k 8644 (0.3 mg/kg, i.p.) was administered prior to a cumulative DZP or PCP dose response determination. RESULTS: None of the VGCC modulators produced robust or consistent alterations in the ethanol-like discriminative stimulus effects of DZP in animals trained with either 1.0 g/kg or 2.0 g/kg ethanol. However, the ethanol like discriminative stimulus effects of PCP were significantly enhanced in the presence of the VGCC antagonists and attenuated in the presence of the agonist in animals trained with 2.0 g/kg ethanol. CONCLUSIONS: Overall, these data show that VGCC modulation is not a robust component of ethanol-like discriminative stimulus effects of DZP in animals trained with 1.0 g/kg or 2.0 g/kg ethanol. However, the ethanol-like effects of PCP, particularly at higher training doses, appear to be modulated by dihydropyridine-sensitive VGCCs. PMID- 10789888 TI - Yohimbine produces antinociception in the formalin test in rats: involvement of serotonin(1A) receptors. AB - RATIONALE: Previous studies have suggested that the alpha2-adrenergic receptor antagonist yohimbine produced antinociceptive effects in the formalin test in rats. However, yohimbine is also an agonist at serotonin (5-HT)1A receptors, suggesting the possibility that the antinociceptive effects of yohimbine might be mediated via these receptors. OBJECTIVE: The purpose of the present studies was to evaluate the potential role of 5-HT(1A) receptors in mediating the antinociceptive effects of yohimbine. METHODS: The antinociceptive effects of yohimbine were evaluated using the formalin test in rats. RESULTS: Yohimbine (2.5 10 mg/kg s.c.) produced dose-related antinociception during both phase I and phase II of the formalin test, and was approximately equipotent and equiefficacious to morphine. The selective 5-HT(1A) receptor antagonist WAY 100,635 (0.03-3.0 mg/kg s.c.) produced a partial reversal of yohimbine. In comparison, the selective 5-HT(1A) receptor agonist (+/-)8-hydroxy dipropylaminotetralin HBr (8OH-DPAT; 1.0 mg/kg s.c.) also produced a dose-related antinociception in the formalin test, although 8OH-DPAT was completely reversed by WAY 100,635 (3.0 mg/kg s.c.). The antinociceptive effects of yohimbine were not antagonized by the 5-HT(1B/1D) antagonist GR 127935 (1.0 mg/kg and 3.0 mg/kg s.c.), the 5-HT2 antagonist LY53857 (1.0 mg/kg s.c.), or the 5-HT3 antagonist zatosetron (3.0 mg/kg s.c.). CONCLUSIONS: The present results demonstrate that yohimbine produces a dose-related antinociception in the formalin test in rats which is mediated in part by the agonistic actions at 5-HT(1A) receptors. PMID- 10789889 TI - State-dependent effects of alcohol on explicit memory: the role of semantic associations. AB - RATIONALE: Memory performance can be facilitated when the context in which retrieval occurs matches the context in which learning initially took place in two separate ways, in form of interactive or independent context. In the present study, the differential effects of alcohol, as independent context, on the free recall of items of high or low semantic associations were investigated. The high and low associations offer different strengths of interactive context at stimulus input, i.e. context that influences what will be stored. METHODS: Using a state dependent retrieval paradigm, alcohol (0.8 g/kg) or placebo was administered prior to encoding and/or retrieval and their effects were tested using measures of free recall. Forty-eight participants were tested according to a traditional state-dependent retrieval design where half of the subjects studied the items under alcohol (A), half under placebo (P) followed by retrieval of the items under A or P giving four groups (AA, AP, PA, PP). RESULTS: Delayed free recall was significantly impaired when alcohol was administered prior to both encoding and retrieval of study material (P<0.05). Alcohol administered prior to encoding and prior to retrieval decreased especially the recall of high association items (P<0.05). Participants in the same-state groups (AA, PP) recalled fewer low association items than participants in disparate state groups (AP, PA; P<0.05). This effect of drug state on low associations may reflect an inability of weaker cues to facilitate retrieval in the presence of stronger cues (i.e. high associations and drug). Indeed, participants in the same-state groups recalled a greater percentage of material in form of high association items than participants in disparate-state groups (P<0.05). CONCLUSIONS: These results demonstrate that alcohol given at encoding and at retrieval specifically impairs retrieval of high association items. However, if the drug state is the same at encoding and at retrieval, the items with the high associations represent a higher proportion of the total recalled items. These data suggest that alcohol may provide an internal context that can facilitate retrieval of information, acquired in semantic context, which otherwise would have been lost. PMID- 10789890 TI - Radiation risks in perspective: radiation-induced cancer among cancer risks. AB - The majority of the public in industrial countries believes that pollution and low doses of radiation are threats to good health. As a matter of fact, when these putative risks are compared to those originating from lifestyle, they appear very small. In particular, the risks associated with low doses of irradiation, even when they are assessed with the most pessimistic models, appear extremely small. Public anxiety is fuelled by the uncertainty regarding the magnitude of this risk and the use of the linear no threshold (LNT) hypothesis, which gives credence to the concept that even the smallest doses are harmful. There are a number of scientific and epidemiological data currently under debate that are not consistent with the LNT hypothesis. For example, no difference in the incidence of cancers or of birth defects has been observed between regions with low or high natural irradiation. This inconsistency between perceptions and data underlines the role of psychological factors studied since 1957 which should be placed in the perspective of the public's present attitude toward risk and technology. Social amplification or attenuation of risk may occur in several ways. Fearful concern about radiation began in 1955, with the beginning of the Cold War, when the possibility of a nuclear holocaust appeared very real. Analysis of the data shows that these fears of technology could have a detrimental effect; they should therefore be investigated and understood. PMID- 10789891 TI - Risk estimates for radiation-induced cancer--the epidemiological evidence. AB - The risk of low-dose radiation exposures has--for a variety of reasons--been highly politicised. This has led to a frequently exaggerated perception of the potential health effects, and to lasting public controversies. A balanced view requires a critical reassessment of the epidemiological basis of current assumptions. There is reliable quantitative information available on the increase of cancer rates due to moderate and high doses. This provides a firm basis for the derivation of probabilities of causation, e.g. after high radiation exposures. For small doses or dose rates, the situation is entirely different: potential increases of cancer rates remain hidden below the statistical fluctuations of normal rates, and the molecular mechanisms of cancerogenesis are not sufficiently well known to allow numerical predictions. Risk coefficients for radiation protection must, therefore, be based on the uncertain extrapolation of observations obtained at moderate or high doses. While extrapolation is arbitrary, it is, nevertheless, used and mostly with the conservative assumption of a linear dose dependence with no threshold (LNT model). All risk estimates are based on this hypothesis. They are, thus, virtual guidelines, rather than firm numbers. The observations on the A-bomb survivors are still the major source of information on the health effects of comparatively small radiation doses. A fairly direct inspection of the data shows that the solid cancer mortality data of the A-bomb survivors are equally consistent with linearity in dose and with reduced effectiveness at low doses. In the leukemia data a reduction is strongly indicated. With one notable exception -- leukemia after prenatal exposure--these observations are in line with a multitude of observations in groups of persons exposed for medical reasons. The low-dose effects of densely ionizing radiations- such as alpha-particles from radon decay products or high-energy neutrons--are a separate important issue. For neutrons, there is little epidemiological information. This has facilitated exaggerated claims of high neutron effects with reference to alleged dangers from transports of reactor fuel. However, in spite of limited information, it can be shown that the data from Hiroshima exclude the stated claims. New dosimetric information on neutrons may turn out to be highly informative with regard to an upper limit for the potential effects of neutrons and equally with regard to a reassessment--and a possible reduction--of risk estimates for gamma-rays. PMID- 10789892 TI - Thyroid cancer risk in Belarus after the Chernobyl accident: comparison with external exposures. AB - Within the time period 1990-1993, childhood thyroid cancer incidence due to the Chernobyl accident increased dramatically in Belarus, especially with regard to the birth cohort January 1, 1971, to May 31, 1986. This rise subsequently slowed down, i.e. during the period 1994-1996. The respective data were analysed and compared with the results of an analysis on the time dependence of thyroid cancer incidence in a pooled cohort of persons who had been exposed during childhood to external radiation with high dose rates. Concerning the period of 5-10 years following exposure, the excess absolute cancer risk per unit thyroid dose in the latter (external) exposure group was found to exceed the one in the Belarus group by a factor of two. This difference, however, is not statistically significant. The age-adjusted average excess absolute risk per unit thyroid dose for the period of 5-50 years following external childhood exposure was found to be 8 female and 14 male cases per 10(4) person-year Gy, which is a factor about 2.5 times higher than for the non-adjusted risk in the pooled cohort, as reported by Ron et al. in 1995. Assessments of future excess thyroid cancer cases due to the Chernobyl accident were done on the basis of the time dependence of thyroid cancer risk following external exposure. The thyroid cancer incidence among the birth cohort considered in Belarus and for a period starting from the cessation of the available observation data (1 January 1997) and extending to 50 years after the Chernobyl accident has been estimated to be about 15,000 cases, with an uncertainty range of 5,000-45,000 cases. According to our calculations, 80% of these cases exceed the baseline risk under enhanced thyroid surveillance. PMID- 10789893 TI - Differential induction of apoptosis in x-irradiated L5178Y sublines bearing p53 mutation. AB - We examined apoptosis and expression of p53, E2F-1, bax, bclx(L) and bc12 proteins in two L5178Y (LY) murine lymphoma sublines, LY-R and LY-S, which differ in radiosensitivity and double-strand break (DSB) repair. Both sublines are heterozygous for a p53 mutation in codon 170 that precludes the transactivation function. Accordingly, there is no G1/S arrest after irradiation. We found that there is no change in expression of E2F-1, bax, bclx(L) or bc12 proteins in both LY sublines after x-irradiation. LY-R cells do not constitutively express bc12, whereas both sublines show high bax content. Radiation induces delayed apoptosis to a greater extent in LY-S than in LY-R cells. The apoptosis can be seen 24 h after irradiation (2 Gy) of LY-S cells, with a maximum at 48 h. LY-R cells need 5 Gy and 72 h post-irradiation incubation to show marked apoptosis (identified by the TUNEL method). The reported observations support the assumption that differential radiosensitivity of LY sublines is associated with the induction of apoptosis that is not related to transactivation by p53 and is primarily related to differential DNA repair ability. PMID- 10789894 TI - Detection of radiation effects using recombinant bioluminescent Escherichia coli strains. AB - Effects of ionizing radiation (0.1-500 Gy) on recombinant Escherichia coli cells containing the stress promoters recA, grpE, or katG, fused to luxCDABE, were characterized by monitoring transcriptional responses reflected by the bioluminescent output. The minimum dose of gamma-irradiation detected by E. coli DPD2794 (recA::luxCDABE) was about 1.5 Gy, while the maximum response was obtained at 200 Gy. The amount of emitted bioluminescence increased proportionally with the gamma-ray doses which were found to elicit a DNA damage response in a range of 1-50 Gy. In addition, the cell growth rate was severely, but transiently, retarded by about 50 Gy. Quantification of the gamma-ray dose may be possible using the recA promoter fusion, since linear enhancement of the bioluminescence emission with increasing gamma-ray dose was observed. Other irradiated strains (50 Gy) responsive to either oxidative stress (DPD2511, katG::luxCDABE) or protein-damaging stress (TV1061, grpE::luxCDABE) did not display an increased bioluminescent output, while DPD2794 irradiated by the same dose of gamma-rays gave a significant bioluminescent output. This indicates that the recA promoter is the one most suitable for developing a biosensor for ionizing radiation. PMID- 10789895 TI - Melanin decreases clastogenic effects of ionizing radiation in human and mouse somatic cells and modifies the radioadaptive response. AB - Melanin's influence on the chromosome aberration frequency induced by radiation in human lymphocytes and mouse bone marrow cells has been studied. We revealed earlier that melanin significantly decreases the frequencies of different radiation-induced mutations in animal germ cells. Melanin protection in somatic cells has been found to be less effective. The melanin effect in somatic cells depends on radiation dose: the lower the damage level, the better the melanin protection. In order to determine the influence of melanin at low radiation doses, the adaptive response was investigated in mouse bone marrow cells in vivo. The level of chromosome aberrations in these cells after fractionated irradiation of 0.2 Gy+1.5 Gy with a 4-h interval was about half that after a single dose of 1.7 Gy. If melanin was injected prior to irradiation, the aberration level decreased by a factor of about two in both cases. This observed result may be due to the potential radioprotective effect of melanin and to the absence of any adaptive response, whereas in the case of melanin application between the priming and challenge doses, the combined effect of the adaptive response as well as melanin protection resulted in a 4-fold decrease of chromosome aberrations. These results allow us to draw the following conclusions: adaptive response can be prevented by a radioprotector such as melanin, and melanin is capable of completely removing low-dose radiation effects. PMID- 10789896 TI - Determination of biokinetic parameters for ingestion of radionuclides of zirconium in animals using stable tracers. AB - Exposure to the radioactive isotope 95Zr, as in nuclear accidents, and to stable zirconium, due to its use in industry, has increased the interest in the biokinetics of this element. Information has been derived mainly from tests performed on animals by means of radioactive tracers. Due to the fact that extrapolation from animals to humans is always open to question, there is an increasing need of a methodology which allows data to be obtained directly from humans. The use of stable tracers, being ethically justifiable, is a powerful tool for providing this information. As two tracers of the same element must be utilized in order to evaluate gut absorption, an analytical technique which is capable of distinguishing and measuring simultaneously different isotopes of zirconium in biological samples is required. Preliminary tests on laboratory animals were performed in order to assess the feasibility of the double tracer technique combined with proton activation analysis. PMID- 10789897 TI - A model of radioiodine transfer to goat milk incorporating the influence of stable iodine. AB - Previously reported models for radioiodine in ruminants cannot account for the effect of variations in stable iodine intake including large countermeasure doses of stable iodine on the transfer of radioiodine to goat milk. A metabolically based model of radioiodine transfer in goats has been parameterised using new experimental data on the effect of countermeasure doses of stable iodine on radioiodine transfer to milk. To account for the effect of dietary stable iodine levels, the model represents the transfer of iodine from the extracellular fluid to milk with Michaelis-Menten kinetics. The model shows good agreement with the experimental data, and the estimated parameters compare favourably with values which can be estimated from the literature. The parameterised model accounts for 95% of the variation in the observed data for milk, faeces, urine and thyroid (n=199). The model has been used to predict the effects of variation in stable iodine intake and the extent of consequent chemical contamination of milk by stable iodine. The time taken for radio-iodine to reach peak concentrations in milk following a deposition event is predicted to vary significantly (ca. 2 days) over a range of expected stable iodine intakes. Doses of stable iodine sufficient to reduce the radioiodine transfer to milk will result in stable iodine concentrations in milk greatly in excess of internationally advised limits. Therefore, we recommend that stable iodine supplementation not be used as a countermeasure to reduce radioiodine transfer to milk. Indeed, model predictions suggest that reductions in stable iodine intake would be a more effective countermeasure. However, this is unlikely to be feasible since the short physical half-life of 131I may not allow adequate time to implement changes in feed manufacture. The model described in this paper is freely available in ModelMaker 3.0 format (http://www.notingham.ac.uk/environmentalmodelling+ ++/). PMID- 10789898 TI - Effect of power frequency harmonics on magnetic field measurements. AB - This paper presents a study of the effect of harmonic frequencies on magnetic field measurements. We introduced magnetic field meters in a known magnetic field of different frequencies: power frequency (50 Hz) as well as 3rd (150 Hz) and 5th (250 Hz) harmonic frequencies. Two magnetic field levels (0.25 A and 2.5 A) were used. A Helmholtz coil was applied to generate an exact magnetic field. The difference between the measurement results at harmonic frequencies and at power frequency was analyzed using the t-test for matched pairs. The test results show significant differences (P< or =0.01) for 13 out of 28 tests carried out, which is probably due to a curved frequency response near the power frequency. It is, therefore, essential to consider harmonic frequencies in magnetic field measurements in practice. PMID- 10789899 TI - 1999 Neuhauser lecture. From a radiologist's judgment to public policy on child abuse and neglect: what have we wrought? PMID- 10789900 TI - Optimization of a fluoroscope to reduce radiation exposure in pediatric imaging. AB - BACKGROUND: A new children's hospital provided the impetus to investigate radiation dose and image quality in a fluoroscope that was specially engineered for pediatric fluoroscopy. Radiation protection management recommends radiation exposures that are as low as reasonably achievable, while still maintaining diagnostic image quality. OBJECTIVES: To obtain comparative phantom imaging data on radiation exposure and image quality from a newly installed fluoroscope before and after optimization for pediatric imaging. MATERIALS AND METHODS: Images were acquired from various thickness phantoms, simulating differing patient sizes. The images were evaluated for visualization of high- and low-contrast objects and for radiation exposure. Effects due to use of the image intensifier anti-scatter grid were also investigated. RESULTS: The optimization of the new fluoroscope for pediatric operation reduced radiation exposure by about 50% (compared to the originally installed fluoroscope), with very little loss of image quality. Pulsed fluoroscopy was able to lower radiation dose to less than 10% of continuous fluoroscopy, while still maintaining acceptable phantom image quality. CONCLUSION: Radiation exposure in pediatric fluoroscopy can be reduced to values well below the exposure settings that are typically found on unoptimized fluoroscopes. Pulsed fluoroscopy is considered a requisite for optimal pediatric fluoroscopy. PMID- 10789901 TI - A multihospital survey of radiation exposure and image quality in pediatric fluoroscopy. AB - BACKGROUND: Traditionally, pediatric radiologists have been advocates of fluoroscopy systems that provide diagnostic images at the lowest possible radiation dose to the pediatric patient. Manufacturers of fluoroscopic equipment vary as to their claims of "low radiation" exposures. OBJECTIVES: To obtain comparative data on radiation exposure and image quality from four pediatric hospitals, across variants of fluoroscopic equipment (such as pulsed versus continuous fluoroscopy). MATERIALS AND METHODS: Images were acquired from phantoms that simulated the size of a 3-year-old child. Phantom results, both stationary and rotating dynamic, were evaluated for radiation exposure and for image resolution of high- and low-contrast objects. RESULTS: Radiation exposure from the four fluoro units varied widely; the lowest-dose selectable fluoro mode produced exposures varying between 34 and 590 mrads/min among the four fluoro units, and the highest-dose selectable fluoro mode produced 540-2,230 mrads/min. The lowest radiation exposures were produced by pulsed fluoro units, and the very lowest radiation exposure was produced by a fluoroscope that had been especially optimized for pediatric imaging. There was only a small variation in image quality among the hospitals for visualization of stationary objects. A wide variability was noted for detection of objects on the moving phantom. CONCLUSIONS: The variability in the number of detected objects was considerably smaller than the variability in radiation exposure. Pulsed fluoroscopy provides improved resolution for moving objects. Optimization of one hospital's fluoroscope especially for pediatric imaging produced the best ratio of image quality to radiation exposure. PMID- 10789902 TI - Congenital short colon with imperforate anus (pouch colon). Report of a case. AB - We report a case of a persistent cloaca and pseudoexstrophy associated with congenital pouch colon in a native-born American female child. This unusual anomaly occurs in two clinical settings. It has been reported in India as an isolated anomaly occurring primarily in males. Pouch colon also occurs in female patients with pseudoexstrophy or closed cloacal exstrophy. The typical anatomic features of this anomaly are discussed. PMID- 10789903 TI - Progressive esophageal leiomyomatosis with respiratory compromise. AB - Leiomyomatosis is a rare neoplastic condition of the pediatric esophagus. Presenting symptoms usually overlap with more common esophageal disorders, namely, gastroesophageal reflux. A patient is presented in whom leiomyomatosis progressed to the point of causing cachexia and respiratory compromise. PMID- 10789904 TI - Malignant peritoneal mesothelioma in two pediatric patients: MR imaging findings. AB - Malignant mesothelioma is a rare tumor in childhood. We present two cases of malignant peritoneal mesothelioma in which contrast-enhanced, fat-saturated magnetic resonance (MR) imaging was used advantageously to detect peritoneal tumor involvement. PMID- 10789905 TI - The posterior approach to pyloric sonography. AB - Overlying bowel gas or gastric distension may occasionally hinder the sonographic diagnosis of hypertrophic pyloric stenosis. To address this problem, a novel approach for obtaining posterior views of the pylorus is reported. Utilizing this approach may decrease the incidence of nondiagnostic pyloric ultrasonography. PMID- 10789906 TI - Intraosseus teratoma of the iliac bone. AB - We present a 4-year-old child with a large iliac bone mass incidentally discovered in a plain abdominal radiograph. The pathological examination revealed a benign mature teratoma. To the best of our knowledge, the occurrence of intraosseous mature teratoma has not been previously reported. The child had had an immature teratoma of the neck discovered in a fetal ultrasound, and resected on day 6 of life. The neck teratoma recurred twice, at 16 months and at 3.5 years of age. In these two recurrences the lesion appeared progressively more mature. At the time of discovery of the iliac bone teratoma there was no evidence of residual neck disease. The radiological and pathological characteristics, differential diagnosis, and clinical course are discussed. PMID- 10789907 TI - MRI appearances of hip abnormalities in mucolipidosis, type III. AB - Mucolipidosis type III (ML-III) is a lysosomal storage disease often presenting with joint involvement. We report the MRI appearance of the hips in two siblings with ML-III showing abnormal signal intensity within the hips with increased synovial thickness. Although the etiology is uncertain this may reflect a fibrous response to ML-III. PMID- 10789908 TI - Use of Doppler ultrasound and 3-dimensional contrast-enhanced MR angiography in the diagnosis and follow-up of post-traumatic high-flow priapism in a child. AB - We report a 7-year-old boy with post-traumatic arterial priapism. Doppler US could not reliably identify or exclude a fistula. MR angiography did not demonstrate an arteriovenous fistula and the child was treated conservatively. The ideal imaging modality should demonstrate the presence or absence of a clinically significant causative lesion which, in high-flow arterial priapism, may need intervention. Three-dimensional, contrast-enhanced MR angiography appears to fulfil these requirements. On the basis of the non-invasive imaging findings, invasive intervention was avoided in this case with a successful outcome. PMID- 10789909 TI - Potential pitfalls in nuclear medicine: a paediatric teaching case. AB - An infant with complex cardiac defects and isomerism had a persistent pyrexia of undetermined aetiology following cardiac surgery. Radionuclide leucocyte scan showed a focus of increased uptake in the right upper quadrant (RUQ) felt to be compatible with an abscess. However, correlation with clinical data and multimodality imaging allowed the correct diagnosis of ectopic splenic tissue in the RUQ to be made. This report emphasises the need for an integrated approach to imaging. Patients with abnormalities of viscero-atrial situs require careful assessment of their splenic status. Visceral heterotaxy may result in confusing imaging appearances unless the anatomy has been accurately documented. PMID- 10789910 TI - Intracranial infantile hemangiopericytoma. AB - Hemangiopericytoma (HP) is a rare vascular tumor that usually occurs in adults, but 10% occur in children. Tumors occurring in the first year of life are even more rare and are referred to as infantile hemangiopericytoma. In this article, we report the imaging, operative, and pathological findings in a patient with an infantile HP. PMID- 10789911 TI - Beware of the ventriculogallbladder shunt. AB - We report a case of distal end malfunction in a child with ventriculoabdominal shunt. The distal placement was in the gallbladder rather than the peritoneal cavity, as is usually the case with a ventriculoperitoneal shunt. Surgeons should be alerted to the possibility of distal terminus of a shunt to be other than the peritoneal cavity, as revision surgery in patients with ventriculogallbladder shunt requires exposure of the metal connector on the gallbladder wall to prevent biliary leakage. PMID- 10789912 TI - Orbital and facial granulocytic sarcoma (chloroma): a case report. AB - Granulocytic sarcoma is an uncommon manifestation of myelogenous leukemias in which focal masses of immature myeloid cells from the granulocytic lineage infiltrate bone and soft tissue. It is most common in the pediatric population and may present at any time in the course of the disease, either concurrently with the onset of leukemia or during a remission or relapse. Occasionally, it may precede the clinical onset of acute myelogenous leukemia (AML), presenting a diagnostic challenge. PMID- 10789914 TI - Neurocutaneous melanosis with hydrocephalus, intraspinal arachnoid collections and syringomyelia: case report and literature review. AB - Neurocutaneous melanosis (NCM) is a rare nonfamilial syndrome, characterised by large or numerous congenital pigmented nevi and excessive proliferation of melanin-containing cells in the leptomeninges. We report the MR findings in the brain and spine of a child with NCM who underwent neurosurgical treatment and was followed up for 8 years. The findings in this child (small hyperintense collections of melanocytes in both temporal lobes, mild meningeal enhancement along the spine and the development of an extensive subarachnoid CSF accumulation with cord compression and syringomyelia) are believed to be exceptionally rare. PMID- 10789913 TI - Pulmonary emboli following therapeutic embolization of cerebral arteriovenous malformations in children. AB - BACKGROUND: Reports of the complicating side effect of pulmonary embolism (PE) following endovascular therapy of cerebral arteriovenous malformations (AVM) in children have been limited in number. Details of its occurrence are yet to be fully elucidated. OBJECTIVE: The hypothesis is that inadvertent pulmonary migration of embolic material is common and may go unrecognized. MATERIALS AND METHODS: Forty-seven patients (ages 1 day to 16 years and 11 months) underwent embolization of a cerebral AVM with at least one material (cyanoacrylate, platinum coils, detachable balloons, polyvinyl alcohol particles). The medical records and chest radiographs were reviewed retrospectively. Chest radiographs were available in 34 patients. The radiographs were analyzed for the presence or absence of foreign material in the lungs. RESULTS: The chest radiographs in 12 patients (35%) showed pulmonary deposits of embolic material; cyanoacrylate in 10 patients and platinum coils in 2. Two of the patients with cyanoacrylate deposits in the lungs developed respiratory distress that required endotracheal intubation. The patients gradually improved after a time period of 7-10 days with conservative treatment. CONCLUSION: PE is not an uncommon complication in children undergoing embolization of brain AVM. Although usually asymptomatic, PE may cause severe symptoms. PMID- 10789915 TI - Pinus pollen in the atmosphere of Vigo and its relationship to meteorological factors. AB - The Pinus genus has an elevated pollen production and an anemophilous nature. Although considered to be hypoallergenic, numerous cases of allergies caused by Pinus pollen have been cited and different authors believe that its allergenicity should be studied in more depth. In the city of Vigo several patients have tested positive for Pinus pollen extracts in skin tests, some of them being mono sensitive to such pollens. In order to ascertain the behaviour of Pinus pollen and its correlation to the main meteorological factors, we carried out an aerobiological study in the city of Vigo from 1995 to 1998 by using a Hirst active-impact volumetric sporetrap, model Lanzoni VPPS 2000, placed on the left bank of the Vigo estuary (42 degrees 14'15"N, 8 degrees 43'30"W). Pinus has high quantitative importance in the airborne pollen spectrum of the city. It is one of the best represented taxa constituting 13%-20% of the total annual pollen levels. The quantity of Pinus pollen present in the atmosphere of the city of Vigo throughout a year is 5,751 grains (as the average for the sampled years), with a very long pollination period, from the middle of January until May. The maximum concentration was recorded in 1998 with 1,105 grains/m(3) on 3 March, a much greater value than those for the previous years. At the end of its pollination period there is usually a final increase in Pinus pollen concentrations coinciding with the pollination of Pinus silvestris, which are more abundant in mountainous areas far from the city. PMID- 10789916 TI - Steady-state balance model to calculate the indoor climate of livestock buildings, demonstrated for finishing pigs. AB - The indoor climate of livestock buildings is of importance for the well-being and health of animals and their production performance (daily weight gain, milk yield etc). By using a steady-state model for the sensible and latent heat fluxes and the CO2 and odour mass flows, the indoor climate of mechanically ventilated livestock buildings can be calculated. These equations depend on the livestock (number of animals and how they are kept), the insulation of the building and the characteristics of the ventilation system (ventilation rate). Since the model can only be applied to animal houses where the ventilation systems are mechanically controlled (this is the case for a majority of finishing pig units), the calculations were done for an example of a finishing pig unit with 1,000 animal places. The model presented used 30 min values of the outdoor parameters temperature and humidity, collected over a 2-year period, as input. The projected environment inside the livestock building was compared with recommended values. The duration of condensation on the inside surfaces was also calculated. PMID- 10789917 TI - Adaptive modification of membrane phospholipid fatty acid composition and metabolic thermosuppression of brown adipose tissue in heat-acclimated rats. AB - Thermogenesis, especially facultative thermogenesis by brown adipose tissue (BAT), is less important in high ambient temperature and the heat-acclimated animals show a lower metabolic rate. Adaptive changes in the metabolic activity of BAT are generally found to be associated with a modification of membrane phospholipid fatty acid composition. However, the effect of heat acclimation on membrane phospholipid fatty acid composition is as yet unknown. In this study, we examined the thermogenic activity and phospholipid fatty acid composition of interscapular BAT from heat-acclimated rats (control: 25+/-1 degrees C, 50% relative humidity and heat acclimation: 32+/-0.5 degrees C, 50% relative humidity). Basal thermogenesis and the total thermogenic capacity after noradrenaline stimulation, as estimated by in vitro oxygen consumption of BAT (measured polarographically using about 1-mm(3) tissue blocks), were smaller in the heat-acclimated group than in the control group. There was no difference in the tissue content of phospholipids between the groups when expressed per microgram of DNA. The phospholipid fatty acid composition was analyzed by a capillary gas chromatograph. The state of phospholipid unsaturation, as estimated by the number of double bonds per fatty acid molecule, was similar between the groups. The saturated fatty acid level was higher in the heat-acclimated group. Among the unsaturated fatty acids, heat acclimation decreased docosahexaenoic acid and oleic acid levels, and increased the arachidonic acid level. The tissue level of docosahexaenoic acid correlated with the basal oxygen consumption of BAT (r=0.6, P<0.01) and noradrenalinestimulated maximum values of oxygen consumption (r=0.5, P<0.05). Our results show that heat acclimation modifies the BAT phospholipid fatty acids, especially the n-3 polyunsaturated fatty acid docosahexaenoic acid, which is possibly involved in the metabolic thermosuppression. PMID- 10789918 TI - Changes in ambient temperature at the onset of thermoregulatory responses in exercise-trained rats. AB - Spontaneous running in a wheel has emerged as a useful method of exercise in rodents. We investigated how exercise training with a running wheel affects ambient temperatures (T(a)) at the onset of thermoregulatory responses in rats. Female rats were allowed to run freely in the wheel for 6 months. Sedentary control rats did not exercise during the same period. After the exercise training period, they were loosely restrained and T(a) values at the onset of tail skin vasodilation and cold-induced thermogenesis were determined by raising or lowering T(a). Resting levels of core temperature and heat production of the exercise-trained rats were significantly higher than those of the controls. T(a) values at the onset of tail skin vasodilation and cold-induced thermogenesis of the exercise-trained rats were higher than those of the controls. The results suggest that, in rats, exercise training with a running wheel elevates ambient temperatures for heat loss and heat production, which may then contribute to maintaining the core temperature at a high level. PMID- 10789920 TI - Complement and the pathogenesis of endotoxic fever. PMID- 10789919 TI - Heat defense control in an experimental heat disorder. AB - Both whole-body heat exposure and intraperitoneal heating (IPH) result in a body temperature (T(b)) fall that occurs once heating is abated ("hyperthermia-induced hypothermia"). This phenomenon involves a decrease in the threshold T(b) (T(b thresh)) for activation of metabolic heat production (cold defense). Whether the T(b-thresh) for ear skin vasodilation (heat defense) also changes during hyperthermia-induced hypothermia remains unknown. In experiment 1, we applied IPH to guinea pigs by perfusing water through a preimplanted intraperitoneal thermode and delivered the total heat load of either approximately 1.5 kJ ("short" IPH; perfusion duration: 14 min) or approximately 3.0 kJ ("long" IPH; 40 min). Short IPH caused skin vasodilation and a 1.1 degrees C rise in T(b); no hypothermia occurred when IPH ceased. Long IPH caused vasodilation and hyperthermia of a comparable magnitude (1.4 degrees C) that were followed by a T(b) fall to 1.9 degrees C below the preheating value. In experiment 2, the Tb-thresh for skin vasodilation was measured twice: at the beginning of long IPH and at the nadir of the post-IPH hypothermia. The two T(b-thresh) values were 39.0 (SEM 0.1)degrees C and 39.2 (SEM 0.2)degrees C respectively. In the controls, the T(b-thresh) was measured at the beginning and after short IPH; both control values were 39.0 (SEM 0.2)degrees C. We conclude that the hyperthermia-induced hypothermia, although previously shown to be coupled with a decrease in the T(b-thresh) for cold defense, occurs without any substantial change in the T(b-thresh) for heat defense. We speculate that postheating thermoregulatory disorders are associated with threshold dissociation, thus representing the poikilothermic (wide dead band) type of T(b) control. PMID- 10789921 TI - Dew measurements along a longitudinal sand dune transect, Negev Desert, Israel. AB - In a desert environment dew can serve as an important source of moisture for plants, biological crusts, insects and small animals. A measurement programme was carried out within a sand dune belt situated in the northwestern Negev desert, Israel, to measure daily amounts of dew deposition as well as micro meteorological conditions during the dew formation and early-morning drying process. Dew quantities were measured by micro-lysimeters along a 200-m transect as well as by the eddy-correlation technique at a reference location. A simple physical model was constructed to simulate the dew deposition process as well as early-morning drying for the interdune area and the north- and south-facing dune slopes. Measurements carried out during September and October 1997 showed that the daily amounts of dew ranged between 0.1 mm/night and 0.3 mm/night within the interdune area. On the slopes, the amounts of dew were about 50% lower. Simulated results agreed well with the field data. PMID- 10789922 TI - Meteorological factors affecting daily urticaceae pollen counts in southwest Spain. AB - The influence of meteorological factors on daily Urticaceae pollen counts were studied in Cordoba (southwest Spain) in 1996 and 1997. The daily Urticaceae pollen concentrations were obtained by using a Hirst-type volumetric sampler, and meteorological data were obtained from the Cordoba airport, located near the sampling site. The highest correlation between pollen concentration and meteorological parameters was obtained during non-rainy seasons. Temperature was found to be the most important meteorological parameter influencing pollen counts in spring, as temperature is the main reason for the increase of pollen concentration in the atmosphere. In autumn, humidity was another important parameter influencing pollen counts. Rain, however, did not appear to be significant. The influence of the pollen concentration of the 2 previous days and the pollen concentration of the previous day has been studied. During periods with low precipitation, the pollen concentration of the previous day was a useful predictor of Urticaceae pollen concentrations for the following day. PMID- 10789924 TI - Pediatric risk of mortality (PRISM) score in meningococcal disease. AB - To assess the pediatric risk of mortality (PRISM) score as a prognostic scoring system in severe meningococcal disease, the files of 53 consecutive patients admitted to a tertiary pediatric intensive care with a clinical diagnosis of meningococcal disease and positive cultures from blood and/or cerebrospinal fluid were analysed. PRISM-score-based expected mortality was compared with observed mortality. Expected mortality in the whole study population was 29% while observed mortality was 19% (P<0.05). The highest expected and observed mortality was found in septicaemic patients without (documented) meningitis, while meningitis patients without septicaemia had the lowest mortality. All patients with a mortality risk below 18.3% (n = 29) survived whereas all those with a mortality risk of 65% or higher (n = 7) died. Of the 17 patients with a mortality risk between 18.3% and 63.9%, 14 survived and 3 died. The area under the receiver operating characteristic (ROC) curve was 0.94, which is at least comparable with the best-performing meningococcal-disease-specific scoring systems. CONCLUSION: The PRISM score is a useful generic measure of severity of illness in meningococcal disease and can be used to determine the effectiveness of different treatment strategies. PMID- 10789923 TI - Prediction of extubation failure in preterm neonates. AB - The aim of this study was to compare the results of lung function measurements made before and after extubation and ventilator settings recorded immediately prior to extubation with regard to their ability to predict extubation success in mechanically ventilated, prematurely born infants. Immediately after extubation all infants were nursed in an appropriate amount of humidified oxygen bled into a headbox. Functional residual capacity, spontaneous tidal volume and compliance of the respiratory system were measured both within 4 h before and within 24 h after extubation. The peak inspiratory pressure and inspired oxygen concentration immediately prior to extubation were recorded. The results were related to extubation failure: requirement for continuous positive airways pressure or re ventilation within 48 h of extubation. A total of 30 infants, median gestational age 29 weeks (range 25-33 weeks) were studied at a median postnatal age of 3 days (range 1-6 days). Extubation failed in ten infants, who differed significantly from the rest of the cohort with regard to their post extubation functional residual capacity (FRC) (median 23, range 15.6-28.7 ml/kg versus 28.6, range 18.1 39.2 ml/kg, P<0.01) and their requirement for a higher inspired oxygen concentration post extubation (median 0.30, range 0.21-0.40 versus 0.22, range 0.21-0.36, P<0.05). An FRC of less than 26 ml/kg post extubation had the highest positive predictive value in predicting extubation failure. CONCLUSION: A low lung volume performed best in predicting extubation failure when compared to the results of other lung function measurements and commonly used 'clinical' indices, i.e. ventilator settings. A low gestational age, however, was a better predictor of extubation failure than a low lung volume. PMID- 10789925 TI - Respiratory distress in a neonate with an enlarged thymus. AB - Thymic hyperplasia, although not a rare condition in infancy, is usually asymptomatic. We describe an infant presenting in the perinatal period with marked tachypnoea. An enlarged thymus, demonstrated on chest radiograph and CT, was associated with small-volume, non-compliant lungs. Other causes of pulmonary malfunction and maldevelopment were excluded. CONCLUSION: Thymic enlargement is unusually associated with neonatal respiratory distress but should be considered in the differential diagnosis. PMID- 10789926 TI - Thrombosis of the deep cerebral veins with excessive bilateral infarction in a premature infant with the thrombogenic 4G/4G genotype of the plasminogen activator inhibitor-1. AB - We report on a preterm infant with deep cerebral venous thrombosis, a rare condition in this age group. This premature infant had a gestational age of 33 weeks and normal development until day 18, when he presented with tonic seizures and a tense fontanelle. Ultrasound and computed tomography revealed bilateral haemorrhagic infarction of the whole region drained by the deep cerebral veins, including the periventricular white matter, thalamus and choroid plexus. The child was homozygous for the 4G allele of the plasminogen activator inhibitor-1 (PAI-1) 4G/5G promoter polymorphism. CONCLUSION: In patients with bilateral cerebral infarction, thrombosis of the deep cerebral veins should be considered. In addition the role of prothrombotic risk factors, including PAI-1 4G/5G promoter polymorphism, in cerebral vein thrombosis should be clarified in a multicentre study. PMID- 10789927 TI - Secondary respiratory chain defect in a boy with long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: possible diagnostic pitfalls. AB - We report on a boy who suffered from microcephaly, growth retardation, cardiomyopathy and hepatic dysfunction. When he had his first febrile infection at the age of 3 months he showed metabolic decompensation. Laboratory parameters and clinical features were compatible with a beta-oxidation defect or a respiratory chain disorder. Measurement of beta-oxidation enzymes showed long chain 3-hydroxyacyl CoA dehydrogenase (LCHAD) deficiency; determination of respiratory chain complex activities revealed complete absence of complex I, II, III and IV activities in skeletal muscle and reduced activities of complexes II and IV in cultured fibroblasts, with secondary dysregulation of ATP synthase. The patient was found to be homozygous for the MTP:G1528 C mutation (LCHAD deficiency). CONCLUSION: This patient had LCHAD deficiency as his primary metabolic disorder, leading to secondary inhibition of respiratory chain enzymes by 'toxic' metabolites. PMID- 10789928 TI - Maternal uniparental disomy 7--review and further delineation of the phenotype. AB - Uniparental disomy (UPD) is defined as the inheritance of both homologous chromosomes from only one parent. So far, maternal UPD 7 has been described in 28 cases. Here, we report 4 new cases, present clinical information of 5 cases previously reported by us, and review the clinical and molecular findings of all 32 cases. We found a phenotype characterized by pre- and postnatal growth retardation, occipitofrontal head circumference in the lower normal range, a triangular face, and retarded bone maturation. Findings of the facial gestalt included a high and broad forehead and a pointed chin. A broad mouth with down turned corners, prominent ears, cafe-au-lait spots, hemihypotrophy, or clinodactyly were rarely present. Psychomotor development was delayed in 6 cases. The clinical findings strikingly resemble the phenotype of the heterogeneous Silver-Russell syndrome (SRS). Other anomalies were less frequently found than in SRS. Molecular investigations revealed 11 cases with isodisomy and 17 cases with heterodisomy. In 4 cases this information was not available. From the allelic distribution of the microsatellites investigated, 9 cases might be the consequence of an error at maternal meiosis I, and 6 cases might be due to non disjunction at maternal meiosis II. Three of the 17 heterodisomic cases had trisomy 7 in chorionic villi, in the remaining cases no prenatal diagnosis through chorionic villus sampling was reported. CONCLUSION: Maternal UPD 7 should investigated in children with pre- and postnatal growth retardation anda facial gestalt characterized by a high and broad forehead and a pointed chin, as well as in cofined placental mosaicism for trisomy 7. PMID- 10789929 TI - Influence of apolipoprotein E genotypes on serum lipid parameters in a biracial sample of children. AB - The goals of this study were to compare the allelic distribution of the apolipoprotein E(apoE) gene in Hungarian and Hungarian Gypsy children and to examine the impact of apoE polymorphism on quantitative levels of lipids in the two racial groups. Our data yielded calculated allele frequencies of 6.4% and 8.9% for apoE2; 83.8% and 73.8% for apoE3; and 9.8% and 17.3% for apoE4 in Hungarian and in Gypsy children, respectively. The frequency of the apoE4 allele was significantly higher (P<0.05) in Gypsy children than in Hungarians. The effect of apoE genotypes on serum lipid parameters differed considerably in the two racial groups. In the Gypsy group the lowest total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and triglyceride levels were in the E3/E3 group and these values differed significantly (P<0.0001 for TC and LDL-C and P<0.01 for triglyceride) from the values in the E2/E3 and E3/E4 groups. There were no significant differences in TC, LDL-C and triglyceride levels between E2/E3 and E3/E4 groups. The high-density lipoprotein-cholesterol (HDL-C) levels did not differ significantly among the genotype groups. In Hungarian children, the apoE2/3 group displayed lower, the E3/4 group higher, values of TC and LDL-C than in the E3/3 group, but the differences were not significant (P>0.05). HDL-C and triglyceride values did not differ among the genotype groups. CONCLUSION: Our results demonstrate that the apolipoprotein E allele frequencies differ between Hungarian and Gypsy children and suggest that these alleles influence the serum lipid levels, but other genetic and environmental factors can considerably change this effect. PMID- 10789930 TI - The incidence of childhood inflammatory bowel disease in Wales. AB - A prospective study was undertaken throughout Wales over the period from 1 January 1995 to 30 March 1997, to determine the incidence of childhood inflammatory bowel disease (IBD). Thirty-eight (24 male) new cases of IBD were reported. Twenty (16 male) of the reported cases had Crohn's disease, 11 (5 male) had ulcerative colitis (UC) and 7 (3 male) indeterminate colitis. Crohn's disease occurred four times more often in boys than girls, whereas there was no sex difference in the incidence for UC and indeterminate colitis. The median age at presentation was 12 (range 1.5-16) years and there was no difference in the age of presentation or the duration of symptoms prior to diagnosis in any of the types of IBD. CONCLUSION: The overall incidence (95% confidence intervals) for IBD in Wales was 2.6 (1.87-3.48) cases per 100,000 per year. The incidence for Crohn's disease was 1.36 (0.86-2.04) cases per 100,000 per year, for UC 0.75 (0.39-1.28) cases per 100,000 per year and for indeterminate colitis 0.48 (0.2 0.92) cases per 100,000 per year. PMID- 10789931 TI - Rib abnormalities arising before and after birth. AB - Rib abnormalities detected in the neonatal period are usually thought to be part of a skeletal disorder. There are, however, many causes which include metabolic bone disease, infection and trauma. Rib abnormalities are also found in general disorders such as chromosomal abnormalities or hypothyroidism. CONCLUSION: A thorough examination of the neonatal chest radiograph must include the review of the skeletal parts, which can often be revealing. PMID- 10789932 TI - Long-term follow-up of portacaval shunt in glycogen storage disease type 1B. AB - In two girls with glycogen storage disease (GSD) type 1b, terminolateral portacaval shunt (PCS) with partial circular resection of the lobus quadratus of the liver was performed at the age of 12 and 10 years, respectively. At that time, the patients had a height of -3.1 and -1.7 SDS, respectively. PCS resulted in a spectacular growth spurt of 35 cm within the first 5 years after surgery in both of them. As first sign of puberty, breast enlargement started 2.5 years after PCS in both patients. Improved glucose tolerance was evidenced by increased levels of blood glucose and insulin after PCS. Diet with raw cornstarch (CS), 2g/kg body weight four times daily, was started 8 years after PCS in patient 1, but initiated with nightly gastric feeding at the age of 2 years in patient 2, 8 years before PCS. Treatment with recombinant granulocyte colony-stimulating factor (rhGCSF), 6 microg/kg body weight every 36-48 h, was started 20 years after PCS in patient 1, but only 1 month before PCS in patient 2. Progressive development of up to 7-8 liver adenomas was observed after PCS, but without conclusive signs of malignancy on Ferrit MRI. The PCS is still open 23 and 7 years after PCS, respectively. Terminolateral PCS with partial circular resection of the lobus quadratus of the liver associated with dietary control and rhGCSF might still have a place in the treatment of GSD type 1b because it improves the tolerance to fasting and the quality of life and moreover yields excellent metabolic control. CONCLUSION: Treatment of glycogen storage disease type 1b by portacaval shunt might be considered in patients with height-for-age below the 3rd percentile occurring in spite of dietary control, or before considering liver transplantation which, if necessary, can still be performed after shunt surgery. PMID- 10789933 TI - Effect of patent ductus arteriosus and indomethacin treatment on serum cardiac troponin T levels in preterm infants with respiratory distress syndrome. AB - Cardiac troponin T (cTnT) represents a sensitive and specific marker of ischemic myocardial damage in adult and neonatal populations. The aim of this study was to detect the potential ischemic effect of persistent patent ductus arteriosus (PDA) and indomethacin treatment on the coronary vascular bed by measuring cTnT concentrations. cTnT levels were measured in 23 preterm infants (<32 weeks of gestational age) with respiratory distress syndrome (RDS), 11 with PDA and 12 without, at 2, 4, and 7 days after birth. cTnT concentrations (mean +/- SEM) significantly decreased (P<0.05) from the 2nd (0.63+/-0.09 microg/l) and the 4th (0.77+/-0.13 microg/l) to the 7th postnatal day (0.28+/-0.04 microg/l). At day 2 after birth, cTnT levels in preterm infants with RDS were significantly higher (P<0.05) than our reference values for healthy preterm neonates (0.63+/-0.09 microg/l vs. 0.18+/-0.04 microg/l). No differences were found between RDS infants with and without PDA at 2 (0.65+/-0.13 vs. 0.61+/-0.14 microg/l), 4 (0.71+/-0.21 vs. 0.87+/-0.16 microg/l), and 7 (0.26+/-0.05 vs. 0.29+/-0.07 microg/l) days of life. In infants with PDA, cTnT levels did not differ before the first dose of indomethacin was given (0.65+/-0.14 microg/l) or 2 h (0.65+/-0.15 microg/l) and 48 h (0.71+/-0.21 microg/l) afterwards. CONCLUSION: In preterm infants with RDS the occurrence of PDA and indomethacin treatment are not associated with ischemic cardiac damage as detected by cTnT measurements. PMID- 10789934 TI - Life-threatening heart failure in meningococcal septic shock in children: non invasive measurement of cardiac parameters is of important prognostic value. AB - Heart failure is a life-threatening complication of fulminant meningococcal septic shock (MSS). Depression of left ventricular function, in particular, is thought to be due to circulating meningococcal endotoxin. Myocardial failure leads to ventricular dilation expressed by an increased left-ventricle end diastolic diameter (LVED). With ultrasonography, LVED can be accurately measured as well as the shortening fraction (SF). In an evaluative study we investigated the accuracy of the SF and compared it to the accuracy of the Glasgow meningococcal septicemia prognostic score (GMSPS) in the prediction of mortality in children with fulminant MSS. In 27 children admitted in a 4-year period with a presumptive clinical diagnosis of fulminant MSS, hypotension persisted for more than 1 h despite volume loading and inotropic therapy. Seven of these children died (26%); all had an SF <0.30 and a GMSPS > or =10 (the sensitivity of both scores was 100%). Positive predictive values of the SF and GMSPS were 41% and 58% respectively. CONCLUSIONS: SF can be used in addition to other severity scores in clinical decision-making and contribute to the selection of children with the worst prospects for inclusion in experimental treatment studies. PMID- 10789935 TI - German cross-cultural adaptation of the Health Utilities Index and its application to a sample of childhood cancer survivors. AB - Steady progress in developing effective treatments for childhood cancer and other severe pediatric diseases has established the need to consider the nature and frequency of late physical and psychological effects. The Health Utilities Index Mark 2 and Mark 3 (HUI2/3) systems were developed by Feeny, Furlong, Torrance et al. in Canada. These systems are generic multi-attribute measures of a person's health status and health-related quality of life. The first German version of the Canadian HUI2/3 questionnaire was created in our clinic, following recommended guidelines for cross-cultural adaptation of health-related quality of life measures. The usefulness of the resultant version was investigated using a sample of 142 patients who presented to our oncological outpatients' department for a routine health care visit after completion of treatment. The 15 items of the HUI2/3-questionnaire were answered independently by three groups of assessors- nurses, physicians, and parents or patients. Two additional questions covered ratings of the severity of treatment effects and the specification of these effects. The questionnaire was both easy to use and acceptable to the assessors. Percentage agreement between observers about levels for individual attributes ranged from 56% to 100%, with the lowest agreement on the subjective attributes of emotion, pain and cognition. These results are in accordance with previous studies using the original instrument. HUI2 global utility scores were significantly related to ratings of treatment sequelae, giving support to the discriminant validity of the measure. CONCLUSION: The German version of HUI2/3 is a useful instrument with generally high inter-observer agreement and good suitability for outcome measurement in childhood cancer patients. Further research is needed to assess the usefulness of the instrument in other clinical populations and its sensitivity in longitudinal studies. PMID- 10789937 TI - Percutaneous closure of patent ductus arteriosus: complementary use of detachable Cook patent ductus arteriosus coils and Amplatzer duct occluders. AB - Several different devices were evaluated for the percutaneous closure of patent ductus arteriosus (PDA), and important drawbacks were found in all of them. To overcome these drawbacks, both detachable Cook PDA coils and Amplatzer duct occluders (ADO) were used for the percutaneous closure of PDA. A total of 54 patients underwent transcatheter occlusion of PDA at a median age of 4.5 years (range 0.5-29 years) and at a median weight of 19.5 kg (range 6-69 kg). Three patients were adults. Detachable Cook PDA coils were used in 26 patients with a median PDA diameter of 1.7 mm (range 1.1-2.2 mm) and ADO were used in 28 patients with a median PDA diameter of 3.8 mm (range 1.9-7.5 mm). Devices were successfully implanted in all 54 patients. Complete closure was achieved in 53 of 54 patients (98% closure rate). Median fluoroscopy time was 12 min (range 4-47 min). CONCLUSION: According to our experience, the complementary use of detachable Cook patent ductus arteriosus coils and Amplatzer duct occluders for the percutaneous closure of PDA can be recommended. PMID- 10789936 TI - Effect on lung function of continuous positive airway pressure administered either by infant flow driver or a single nasal prong. AB - The aim of this study was to assess if continuous positive airways pressure (CPAP) delivered by an infant flow driver (IFD) was a more effective method of improving lung function than delivering CPAP by a single nasal prong. A total of 36 infants (median gestational age 29 weeks, range 25-35 weeks) were studied, 12 who received CPAP via an IFD, 12 who received CPAP via a single nasal prong and 12 without CPAP. CPAP was administered post extubation if apnoeas and bradycardias or a respiratory acidosis developed or electively if the infant was of birth weight <1.0 kg. Lung function was assessed by the supplementary oxygen requirement and measurement of compliance of the respiratory system using an occlusion technique. Assessments were made immediately prior to and after 24 h of CPAP administration and at similar postnatal ages in the non-CPAP group. The infants who did not require CPAP had better lung function (non significant) than the other two groups before they received CPAP. After 24 h, lung function had improved in both CPAP groups to the level of the non CPAP infants. The supplementary oxygen requirements of all three groups decreased over the 24 h period, but this only reached significance in the single nasal prong group (P<0.05). Four infants supported by the IFD, but none with a single nasal prong, became hyperoxic. CONCLUSION: Continuous positive airways pressure administration via the infant flow driver appears to offer no short-term advantage over a single nasal prong system when used after extubation in preterm infants. PMID- 10789938 TI - A 7-year-old boy with dextrocardia and dysphagia. PMID- 10789939 TI - Does growth hormone therapy improve motor development in infants with Prader Willi syndrome? PMID- 10789940 TI - Improvement of peripheral neuropathy with oral prednisolone in Chediak-Higashi syndrome. PMID- 10789941 TI - Convergence of posterior semicircular canal and saccular inputs in single vestibular nuclei neurons in cats. AB - Convergence between posterior canal (PC) and saccular (SAC) inputs in single vestibular nuclei neurons was investigated in decerebrated cats. Postsynaptic potentials were recorded intracellularly after selective stimulation of the SAC and PC nerves. Stimulation of either the SAC or PC nerve orthodromically activated 143 vestibular nuclei neurons. Of these, 61 (43%) were antidromically activated by stimulation of the C1-C2 junction, 14 (10%) were antidromically activated by stimulation of the oculomotor or trochlear nucleus, and 14 (10%) were antidromically activated by stimulation of both the oculomotor or trochlear nucleus and the spinal cord. Fifty-four (38%) neurons were not activated by stimulation of either or both. We named these neurons vestibulospinal (VS), vestibulo-ocular (VO), vestibulooculo-spinal (VOS) and vestibular (V) neurons, respectively. Both PC and SAC inputs converged in 47 vestibular nuclei neurons (26 VS, 2 VO, 6 VOS and 13 V neurons). Of these, 19 received monosynaptic excitatory inputs from both nerves. This input pattern was frequently seen in VS neurons. Approximately half of the convergent VS neurons descended to the spinal cord through the lateral vestibulospinal tract. The remaining half and all the convergent VOS neurons descended to the spinal cord through the medial vestibulospinal tract. Most of the convergent neurons were located in the lateral nucleus or descending nucleus. PMID- 10789942 TI - Sacculo-ocular reflex connectivity in cats. AB - The otolith system contributes to the vestibulo-ocular reflexes (VOR) when the head moves linearly in the horizontal plane or tilts relative to gravity. The saccules are thought to detect predominantly accelerations along the gravity vector. Otolith-induced vertical eye movements following vertical linear accelerations are attributed to the saccules. However, information on the neural circuits of the sacculo-ocular system is limited, and the effects of saccular inputs on extraocular motoneurons remain unclear. In the present study, synaptic responses to saccular-nerve stimulation were recorded intracellularly from identified motoneurons of all twelve extraocular muscles. Experiments were successfully performed in eleven cats. Individual motoneurons of the twelve extraocular muscles--the bilateral superior recti (SR), inferior recti (IR), superior obliques (SO), inferior obliques (IO), lateral recti (LR), and medial recti (MR) were identified antidromically following bipolar stimulation of their respective nerves. The saccular nerve was selectively stimulated by a pair of tungsten electrodes after removing the utricular nerve and the ampullary nerves of the semicircular canals. Stimulus intensities were determined from the stimulus-response curves of vestibular N1 field potentials in order to avoid current spread. Intracellular recordings were performed from 129 extraocular motoneurons. The majority of the neurons showed no response to saccular-nerve stimulation. In 17 (30%) of 56 extraocular motoneurons related to vertical eye movements (bilateral SR and IR), depolarizing and/or hyperpolarizing postsynaptic potentials (PSPs) were observed in response to saccular-nerve stimulation. The latencies of PSPs ranged from 2.3 to 8.9 ms, indicating that the extraocular motoneurons received neither monosynaptic nor disynaptic inputs from saccular afferents. The majority of the latencies of the depolarization, including depolarization-hyperpolarization, were in the range of 2.3-3.3 ms. Latencies of hyperpolarizations were typically longer than those of depolarizations. Only one contralateral SO motoneuron of 43 recorded oblique extraocular motoneurons (bilateral SO and IO) showed a depolarization-hyperpolarization in response to saccular-nerve stimulation at a latency of 2.5 ms. None of 30 recorded horizontal extraocular motoneurons (bilateral LR and MR) responded to stimulation of the saccular nerve. The neural linkage in the sacculo-ocular system is relatively weak in comparison to the utriculo-ocular and sacculo-collic systems, suggesting that the role of the sacculo-ocular system in stabilizing eye position may be reduced when compared with utriculo-ocular and semi-circular canal-ocular reflexes. PMID- 10789943 TI - Role of the pretectal nucleus of the optic tract in short-latency ocular following responses in monkeys. AB - When a large-field image is suddenly moved in front of an observer, an ocular following response (OFR) with short latency (<60 ms in monkey and <85 ms in human) is observed. Previous studies have shown that neurons in the pretectal nucleus of the optic tract (NOT) of the monkey respond to movements of large field visual stimuli. To understand the potential role of the NOT in the OFR, we first recorded single-unit activity in the NOT of four monkeys (Macaca fuscata). Sixty-six NOT neurons preferred large-field ipsiversive visual motion. In 86% (49/57) of the neurons, optimal directions were distributed over +/-30 degrees from ipsilateral. NOT units were sensitive to the speed of the visual motion; 54% (27/50) preferred slow (< or =20 degrees/s), 22% (11/50) preferred fast (> or =80 degrees/s) and the remainder intermediate speeds. Their response latencies to the moving visual scene were very short (approximately 51 ms), and 44% of them led the onset of the OFR by 10 ms or more. To characterize the response properties of these neurons, we reconstructed the temporal firing patterns of 17 NOT neurons, using the acceleration, velocity, position and bias components of retinal image slip or eye movements during the OFR by a least squares error method. For each stimulus speed fitting condition, using either retinal slip or eye movements, their firing patterns were matched to some extent although the goodness of fit was better using retinal slip than when eye movements were used. Neither of these models could be applied independently of stimulus speed, suggesting that the firing pattern of the NOT neurons represented information associated with retinal slip or eye movements during the OFR, over a limited range. To provide further evidence that the NOT is involved in generating the OFR, we placed unilateral microinjections of muscimol into the NOT. Following the muscimol injection, we observed a approximately 50% decrease in eye velocity of the OFR toward the side of injection regardless of stimulus speed, while only a weak effect was observed in the OFR during contraversive or vertical image motion. These results suggest that the NOT may play a role in the initiation and support of the short-latency ocular following response. PMID- 10789944 TI - Ionotropic glutamate receptor-mediated responses in the rat primary somatosensory cortex evoked by noxious and innocuous cutaneous stimulation in vivo. AB - To examine the involvement of different ionotropic glutamate receptors in the mediation of responses evoked by noxious cutaneous stimulation, single unit recordings were made from 31 neurons in the primary somatosensory (SI) cortex of rats anesthetized with urethane. To compare synaptic receptor pharmacology across somatosensory submodalities, 13 of the neurons were also tested with an innocuous, cutaneous air jet stimulus. Mechanical (HT) responses, evoked by a 5-s noxious pinch, decayed gradually upon termination of the stimulus and lasted on average for 15.1+/-1.9 s (+/-SEM; n=10). An increase in baseline activity was also observed during noxious stimulus trials of 5-min stimulus intervals. A correlation between increase in mechanical or thermal HT responses and baseline activity was found for some neurons. However, the normalized ratios of the mechanical or thermal HT response to baseline activity during iontophoretic application of (RS)-3-(2-carboxypiperazine-4-yl)-propyl-l-phosphonic acid (CPP), an N-methyl-D-aspartic acid (NMDA) receptor antagonist (0.6+/-0.1; n=11, or 6 nitro-7-sulfamoylbenz[f]quinoxaline-2,3-dione (NBQX), an (RS)-alpha-amino-3 hydroxy5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor antagonist (0.8+/-0.1; n=11), suggest that the reductions in baseline activity did not account for the reductions of the mechanical or thermal HT responses observed, which were reduced proportionally more than the baseline activity. A 10-ms air jet evoked a biphasic increase in action potentials above an average background activity of 7+/-2 spikes/s (n=13). The early phase of this low-threshold (LT) response was within two or three 10-ms bins and had an average firing rate of 74+/-11 spikes/s evoked in the first 10-ms bin (n=13). In eight neurons, the early LT response was followed by a lower frequency excitatory component lasting an average of 415+/-92 ms. Iontophoretic application of CPP reduced responses evoked by a noxious pinch (21+/-10% of control responses; n=19) and a noxious thermal stimulus (24+/-18%; n=5). The fast component of the LT responses was only reduced to 85+/-4% (n=12). A slower component of the LT responses, when present, was also reduced by CPP (15+/-19%; n=4). Iontophoretic application of NBQX reduced responses evoked by a noxious pinch (42+/-12%; n=19) and a noxious thermal stimulus (63+/-16%; n=8). The fast component of the LT responses was reduced to 43+/-6% (n=12) and the slower component to 32+/-20% (n=6). These data show that both NMDA and AMPA/kainate receptors are involved in the mediation of SI high-threshold responses. This same combination of glutamate receptors also mediates low-threshold synaptic responses. PMID- 10789945 TI - Characteristics of sequential movements during early learning period in monkeys. AB - We previously demonstrated that the organization of a learned sequential movement, after long-term practice, is based on the entire sequence and that the information pertaining to the sequence is largely specific to the hand used for practice. However, it remained unknown whether these characteristics are present from the beginning of learning. To answer the question, we examined the performance of four monkeys for the same sequential procedure in the early stage of learning. The monkeys' task was to press five consecutive pairs of buttons (which were illuminated), in a correct order for every pair, which they had to find by trial-and-error during a block of trials. We first examined whether the memory of a sequential procedure that was learned once was specific to the hand used for practice. The second time that the monkeys attempted to learn a novel sequence, they were required to use either the same hand they used the first time or the opposite hand. The number of errors decreased to a similar degree in the same-hand condition and in the opposite-hand condition. The performance time decreased in the same-hand condition, but not in the opposite-hand condition. The results suggest that, in the early stage of learning, memory of the correct performance of a sequential procedure is not specific to the hand originally used to perform the sequence (unlike the well-learned stage, where the transfer was incomplete), whereas memory of the fast performance of a sequential procedure is relatively specific to the hand used for practice (like the well-learned stage). We then examined whether memory of a sequential procedure depends on the entire sequence, not individual stimulus sets. For the second learning block, we had the monkey learn the sequence in the same or reversed order. In the reversed order, the order within each set was identical, but the order of sets was reversed. The number of errors decreased in both the same-order and reversed-order conditions to a similar degree for two out of four monkeys; the decrease was larger in the same-order condition for the other two monkeys. For all monkeys, the performance time decreased in the same-order condition, but not in the reversed-order condition. The results suggest that the memory structure for correct performance varies among monkeys in the early stage of learning (unlike the well-learned stage, where the memory of individual sets was consistently absent). On the other hand, memory of the fast performance of a sequential procedure is relatively specific to the learned order used for practice (like the well-learned stage). PMID- 10789946 TI - Deficits in the coordination of multijoint arm movements in patients with hemiparesis: evidence for disturbed control of limb dynamics. AB - This study provides a detailed analysis of disturbances in the kinematics and dynamics of the acceleration phase of multijoint arm movements in six patients with chronic hemiparesis. Movements of the dominant and nondominant limbs were also examined in three control subjects. Subjects performed rapid movements from a central starting point to 16 targets located equidistantly around the circumference of a circle. Support of the upper limb was provided by an air bearing apparatus, which allowed very low friction movements in the horizontal plane. We found that patients retained the capacity to modulate, in response to target direction, the initial direction of movements performed with the paretic limb. However, in comparison to the nonparetic limb or control subjects, movements of the paretic limb were misdirected systematically. An inverse dynamics analysis revealed an abnormal spatial tuning of the muscle torque at the elbow used to initiate movements of the paretic limb. Based on electromyographic recordings, similar spatial abnormalities were also apparent in the initial activations of elbow muscles. We argue that these spatial abnormalities result from a systematic disturbance in the control signal to limb muscles that cannot be attributed to previously identified mechanisms such as weakness, spasticity mediated restraint, or stereotypic muscle activation patterns (muscle synergies). Instead, our analysis of movement dynamics and simulation studies demonstrate that the spatial abnormalities are consistent with an impaired feedforward control of the passive interaction torques which arise during multijoint movements. This impaired control is hypothesized to reflect a degradation of the internal representation of limb dynamics that occurs either as a primary consequence of brain injury or secondary to disuse. PMID- 10789947 TI - Effects of excitotoxic lesions in the ventral striatopallidal--thalamocortical pathway on odor reversal learning: inability to extinguish an incorrect response. AB - The role of the ventral striatopallidal pathway and related cortical areas in stimulus-reward association reversal behavior was studied by inducing bilateral lesions with the excitotoxin, N-methyl-D-aspartate (NMDA) at restricted sites in the system. The areas lesioned were the ventral pallidum (VP), the ventral striatum (VS), the medial prefrontal cortex (mPFC) [i.e., the prelimbic (PL) and infralimbic (IL) cortexes], and the orbital cortex [i.e., the dorsolateral orbital (DLO), ventral lateral orbital (VLO), and lateral orbital (LO) cortexes]. Rats with lesions of the dorsal caudate nucleus and putamen (CPu) served as a positive control in this study. Water-deprived rats were trained on a go, no-go two-odor olfactory discrimination task to respond to one odor (S+) with water as a reward and to suppress responding to the other odor (S-). The rats were then tested for their ability to reverse the associated stimuli. The number of errors made before successfully learning the stimulus-reward association were measured in relation to a sham lesion control group which did not receive injections of NMDA. In experimental rats, the lesions did not affect their ability to learn stimulus-reward associations for novel odors, but did result in an increase in the number of false alarms after the significance of the associated stimuli were reversed. That is, the lesioned animals persisted in responding to the formerly rewarded but now unrewarded stimulus. Rats with damage to the CPu did not show a significant effect when compared with the controls during reversal problems. The results support the hypothesis that the ventral striatopallidal system, together with related thalamic and frontal cortical structures, functions in reversal learning by suppressing inappropriate responses to stimuli that are no longer rewarded. PMID- 10789948 TI - Superposition of independent units of coordination during pointing movements involving the trunk with and without visual feedback. AB - Previous studies addressing the problem of the control of multiple degrees of freedom have examined the influence of trunk movement on pointing movements within the arm's reach. Such movements may be controlled by two functionally independent units of coordination (synergies): one involving only arm joints and producing the hand trajectory to the target (the transport synergy), and the other coordinating trunk and arm movements leaving the hand trajectory unchanged (the compensatory synergy). The question of whether or not this functional subdivision depends on visual feedback was addressed in the present study. We also tested whether or not the motor effects of different synergies are summated as independent components, a control strategy called "superposition." Finally, we investigated whether or not the relationship between different degrees of freedom within each synergy could be considered linear resulting in proportional changes in different joint angles. Seated subjects produced fast, uncorrected arm movements to an ipsi- or a contralateral target in the direction of +/-45 degrees to the sagittal midline of the trunk. Targets could be reached using the arm alone (control trials) or by combining the arm motion with a forward or backward trunk motion produced by hip flexion or extension (test trials), with and without visual feedback. The shape of the hand trajectory, its direction and tangential velocity, movement precision, joint angles and the sequence of the trunk and hand recruitment and de-recruitment were measured. In both visual conditions, the direction of the hand trajectory observed in control trials was generally preserved in test trials. In terms of sequencing, even in the absence of vision, the trunk movement was initiated before the onset of and outlasted the hand shift, indicating that the potential influence of the trunk on the hand movement was compensated by rotations in the elbow and shoulder joint. The analysis of other variables also implied that the effects of trunk recruitment on the hand trajectory were minor compared to those which could be observed if these effects were not compensated by appropriate changes in the arm joint angles. It was concluded that an arm-trunk compensatory synergy is present in pointing movements regardless of visual feedback. Principal component analysis showed that the relationship between elbow, shoulder and hip joint angles in individual arm and combined arm-trunk movements cannot be considered linear, implying that this relationship is adjusted according to the changing arm geometry. The changes in each arm joint angle (elbow, shoulder) elicited by a forward trunk bending in one block of trials were compared with those elicited by a backward bending in another block, whereas the hand moved to the same target in both blocks. These changes were opposite but of similar magnitude. As a result, for each moment of movement, the mean joint angle obtained by averaging across two directions of trunk motion was practically identical to that in control trials in which the trunk was motionless. It is concluded that the transport and arm-trunk compensatory synergies are combined as independent units, according to the principle of superposition. This principle may simplify the control of the coordination of a redundant number of degrees of freedom. PMID- 10789949 TI - Optokinetic eye movements elicited by an apparently moving visual pattern in guinea pigs. AB - Based on known anatomical and physiological properties, guinea pigs could be expected to show a lack of cortical impact on the basic properties of optokinetic nystagmus (OKN) such as eye movements elicited by a rotating striped drum. The present study aimed to clarify the question of whether the guinea pig's brainstem mediated circuit is capable of generating horizontal OKN with the animal binocularly viewing a stationary stroboscopically illuminated striped pattern (apparent-motion OKN). A striped drum with a pattern periodicity of 2.37 degrees was rotated around the animal. The OKN buildup under real stimulation conditions was found to be largely devoid of a rapid initial rise in slow-phase eye velocity (direct OKN component). Only in a few recordings was a slight initially faster acceleration seen eliciting slow-phase eye velocities of 2 degrees/s at the most. The lack of a rapid initial rise proves that the optokinetic reflex loop does not possess a functionally significant cortically mediated component. To achieve apparent-motion OKN, initially in each recording real-motion OKN was elicited by the constantly illuminated rotating pattern. Then, 60 or 90 s later the drum was stopped and the illumination switched over from constant to flashing light. Following Grusser and Behrens (1979), we calculated an apparent stimulus velocity (= flash frequency x 2.37 degrees) and an apparent OKN gain. We found that all animals continued in nystagmus under the flashing light condition. The slow-phase eye velocity was the same as under real pattern motion, with gain values of between 0.36 and 0.7. In two animals we produced an apparent gain close to unity by suddenly doubling the flash interval, i.e., halving the apparent drum velocity. This gain, however, did not remain at a high level: The slow-phase eye velocity decreased until the previous lower gain was reached again. The data indicate that cortical mechanisms are not necessary to produce nystagmus under apparent-motion stimulation and neither phi nor sigma phenomena of motion perception satisfactorily explain the apparent-motion OKN in the guinea pig. Our data provide evidence that the brainstem-located velocity storage mechanism plays a decisive role in keeping the eye in motion under apparent-motion stimulation. PMID- 10789950 TI - Detecting motor abnormalities in preterm infants. AB - As a consequence of the fragility of various neural structures, preterm infants born at a low gestation and/or birthweight are at an increased risk of developing motor abnormalities. The lack of a reliable means of assessing motor integrity prevents early therapeutic intervention. In this paper, we propose a new method of assessing neonatal motor performance, namely the recording and subsequent analysis of intraoral sucking pressures generated when feeding nutritively. By measuring the infant's control of sucking in terms of a new development of tau theory, normal patterns of intraoral motor control were established for term infants. Using this same measure, the present study revealed irregularities in sucking control of preterm infants. When these findings were compared to a physiotherapist's assessment six months later, the preterm infants who sucked irregularly were found to be delayed in their motor development. Perhaps a goal directed behaviour such as sucking control that can be measured objectively at a very young age, could be included as part of the neurological assessment of the preterm infant. More accurate classification of a preterm infant's movement abnormalities would allow for early therapeutic interventions to be realised when the infant is still acquiring the most basic of motor functions. PMID- 10789951 TI - The subdominant hand increases in the efficacy of voluntary alterations in bimanual coordination. AB - Subjects performed a bimanual circle-tracing task in time with an auditory metronome while restricted to moving with either proximal or distal musculature of the upper limb. Patterns were made in symmetric or asymmetric directions with respect to the midline. Symmetric patterns were more stable than asymmetric patterns. In response to a visual stimulus, subjects reversed the direction of one limb. Unwanted disruptions (momentary or lasting reversals) in the limb contralateral to the reversing limb were observed in 48% of trials. Incidence of disruption was equivalent between postures, but occurred more frequently when the dominant hand reversed direction. This result is consistent with anisotropy in coupling between hands and reveals a unifying constraint between spontaneous and intentional dynamics in bimanual coordination. PMID- 10789952 TI - Nonspecific velocity and amplitude hunting reveals different specific detection performance of flexion and extension movements in the human knee joint. AB - A debate exists in the literature as to what extent perception is conscious. In some publications regarding proprioception, the term proprioception has explicitly only been seen as properly used when subjects were able to report the imposed movement's direction. Detections of movements without movement-direction perception, have been seen as nonspecific. Since a lot of studies discussed this point but never tested it explicitly, we tested it by using nonspecific hunting paradigms (only the occurrence of a movement has to be detected, not its direction) with the following rationale. If the perception performance is really nonspecific, no difference regarding the movement's direction should be found. Thus, if we found a different detection performance regarding flexion and extension by means of a nonspecific paradigm, it would demonstrate that this "nonspecific" perception is already specific. Therefore, we measured the perception characteristics separately for flexion and extension. The perception characteristics have been determined from the amplitude-velocity-relation curve. Two different, nonspecific hunting paradigms, modifications of our previously published specific hunting paradigm, have been used. Thus, we determined pairs of threshold values, regarding amplitude hunting using three different angular velocities (0.1 degrees s(-1), 0.25 degrees s(-1), and 0.5 degrees s(-1)) and regarding velocity hunting using three different angular displacements (0.25 degrees, 0.5 degrees, and 1 degrees) as well, for flexion and extension, respectively. We found that both threshold paradigms (velocity hunting and amplitude hunting) revealed the same perception characteristics for a given movement direction. With an increasing angular velocity, angular displacement threshold values converged toward a common value for flexion and extension (about 0.2 degrees); with an increasing angular displacement, angular velocity threshold values converged toward separate values for flexion (about 0.06 degrees s(-1)) and extension (about 0.1 degrees s(-1)). Thus, our findings demonstrate that detection performance is specific and not bound to conscious perception, since specific thresholds for flexion and extension were revealed with nonspecific hunting paradigms. PMID- 10789953 TI - Selective lesions of the entorhinal cortex, the hippocampus, or the fimbria fornix in rats: a comparison of effects on spontaneous and amphetamine-induced locomotion. AB - Using adult Long-Evans male rats, this experiment compared spontaneous (assessed 15 days and 4.5 months after surgery) and amphetamine-induced (assessed from 4.5 months after surgery onwards; 1 mg/kg, i.p., ten injections, 48 h apart) locomotor activity following N-methyl-D-aspartate lesions of the entorhinal cortex, electrolytic lesions of the fimbria-fornix, or ibotenate lesions of the hippocampus. Sham-operated rats were used as controls. Hippocampal and fimbria fornix lesions, but not entorhinal-cortex lesions induced diurnal and nocturnal hyperactivity, which was attenuated over time, but only in rats with fimbria fornix lesions. Amphetamine-induced hyperlocomotion was assessed in a familiar environment. Lesions of the entorhinal cortex potentiated the locomotor effects of amphetamine, but not lesions of the hippocampus or interruption of the axons in the fimbria-fornix pathway. Sensitization appeared to be decreased by fimbria fornix lesions and to be prevented by hippocampal lesions. Rats with entorhinal cortex lesions behaved as if they had already been sensitized by the lesion. These results clearly show that lesions of the fimbria-fornix, the hippocampus, and of the entorhinal cortex have different effects on spontaneous and amphetamine-induced hyperactivity, as they also have on learning and memory tasks. PMID- 10789954 TI - Cachexia--quo vadis? PMID- 10789955 TI - Report from Spain. AB - Palliative care programmes have expanded enormously in recent years in Spain, with great success in terms of coverage of the target population and opioid consumption, but at the expense of deficiencies in the training of professionals and scarcity of fully comprehensive palliative care systems. A balance has developed with an almost even distribution of resources between home care and in patient services, in spite of a strong tendency in recent years to prioritize domiciliary care in the wake of policies requiring limitation of health expenditure. PMID- 10789956 TI - The symptoms of advanced cancer: relationship to age, gender, and performance status in 1,000 patients. AB - A multivariate analysis of the data was conducted to evaluate the effects of age, gender, and performance status on symptom profile. A comprehensive prospective analysis of symptoms was conducted in 1,000 patients on initial referral to the Palliative Medicine Program of the Cleveland Clinic. The median number of symptoms per patient was 11 (range 1-27). The ten most prevalent symptoms were pain, easy fatigue, weakness, anorexia, lack of energy, dry mouth, constipation, early satiety, dyspnea, and greater than 10% weight loss. The prevalence of these 10 symptoms ranged from 50% to 84%. Younger age was associated with 11 symptoms: blackout, vomiting, pain, nausea, headache, sedation, bloating, sleep problems, anxiety, depression, and constipation. Gender was associated with 8 symptoms. Males had more dysphagia, hoarseness, >10% weight loss and sleep problems; females, more early satiety, nausea, vomiting, and anxiety. Performance status was associated with 14 symptoms. Advanced cancer patients are polysymptomatic. Ten symptoms are highly prevalent. Symptom prevalence for 24 individual symptoms differs with age, or gender, or performance status. PMID- 10789957 TI - Cachexia and anorexia: cancer's covert killer. AB - Cachexia and anorexia are often not observed at the time of diagnosis of cancer. While the initial medical intervention for cancer patients includes antitumor therapy and pain management, the consequences of cachexia and anorexia may be ignored, to the detriment of the patient's quality of life and his or her potential response to chemotherapy. The importance of a well-defined therapeutic strategy to treat cachexia is in order if the patient's overall wellbeing is to improve. Presented is a review of the pharmacological management of anorexia and cachexia, including a four-step ladder approach to medical management. PMID- 10789958 TI - Comparison of octreotide and hyoscine butylbromide in controlling gastrointestinal symptoms due to malignant inoperable bowel obstruction. AB - In advanced cancer patients with inoperable bowel obstruction, the administration of antisecretive and antiemetic drugs has proved to be effective in controlling gastrointestinal symptoms caused by bowel obstruction. However, controlled studies concerning the most effective antisecretive drug are lacking. The aim of this randomized controlled study was to determine whether octreotide or hyoscine butylbromide was the more effective antisecretive drug for use in states of inoperable bowel obstruction. Eighteen patients with inoperable bowel obstruction randomly received octreotide 0.3 mg daily (n = 9) or hyoscine butylbromide (HB) 60 mg daily (n = 9) s.c. The following parameters were measured: episodes of vomiting, nausea, drowsiness, continuous and colicky pain, using a Likert scale corresponding to a numerical value: (none 0, slight 1, moderate 2, severe 3) recorded before starting the treatment (T0) and 24 h (T1), 48 h (T2) and 72 h after (T3), and the mean daily amounts of fluids administered i.v. or s.c. during the period of study. Three patients dropped out of the study because data were incomplete. Octreotide treatment induced a significantly rapid reduction in the number of daily episodes of vomiting and intensity of nausea compared with HB treatment at the different time intervals examined. No relevant changes were found in dry mouth, drowsiness and colicky pain. Lower levels of hydration were associated with nausea regardless of the treatment. At the doses used in this study, octreotide was more effective than HB in controlling gastrointestinal symptoms of bowel obstruction. Further studies are necessary to understand the role of hydration more clearly in such a clinical situation. PMID- 10789959 TI - A prospective study on hyponatraemia in medical cancer patients: epidemiology, aetiology and differential diagnosis. AB - The frequency of hyponatraemia varies from less than 1% to more than 40% in series reported from general hospitals. We performed a prospective study to evaluate its incidence and to determine the types of hyponatraemia in a dedicated cancer hospital. All patients admitted to the Department of Medicine were prospectively studied over 11 months. Hyponatraemia was defined as a serum Na level < or =130 mEq/l. Urine and blood samples were collected at baseline, and all consecutive hyponatraemic episodes were studied. One hundred and six patients developed 123 episodes of hyponatraemia. The observed incidence of hyponatraemia was 3.7%. Sodium depletion and syndrome of inappropriate antidiuretic hormone secretion (SIADH) each accounted for almost one third of all aetiologies. Serum urea and uric acid, urinary Na and fractional excretions of Na and urea were most useful for the differential diagnosis. The percentage of deaths observed in the hyponatraemic group, 19.5%, was higher than in the whole cancer population (6.3%) although no death was apparently due directly to hyponatraemia. Hyponatraemia is regularly diagnosed in cancer patients, but it was related to SIADH in only about one third of the cases. A higher mortality was observed in hyponatraemic patients than in normonatraemic patients. PMID- 10789960 TI - Low-dose continuous-infusion ceftazidime monotherapy in low-risk febrile neutropenic patients. AB - One hundred and thirty-five cancer patients admitted with low-risk neutropenic fever received a low-dose schedule of ceftazidime as infusional monotherapy over a total of 180 episodes. Ceftazidime was administered as a 1-g bolus followed by a continuous infusion of 2 g per day. In this patient population the ceftazidime was both practical and well tolerated. Sixty-eight percent of patients responded with clinical improvement and complete resolution of fever within 48 h. Overall, 95% of patients responded, although 18% subsequently required antibiotic modification for persistent fever. Only 5% of episodes were considered failures due to clinical deterioration, and over the study period there was only 1 fatality due to respiratory failure. The median duration of hospitalisation was only 4 days (2-20). In conclusion, monotherapy with low-dose infusional ceftazidime appears safe and highly effective in this low-risk population of neutropenic patients and may reduce antibiotic costs appreciably. PMID- 10789961 TI - A phase II study of Biotene in the treatment of postradiation xerostomia in patients with head and neck cancer. AB - One of the major side effects of radical radiation therapy for head and neck malignancies is xerostomia, or dryness of the mouth. There is no clearly effective treatment for this condition, but we have observed that patients in our practice believe that their symptoms improve significantly when using two "over the-counter" oral comfort products - Biotene (toothpaste, mouthwash and chewing gum) and Oralbalance gel. We decided to study these agents in a formal phase II study to evaluate their usefulness in patients with postirradiation xerostomia. Twenty-eight patients with post-irradiation xerostomia were entered on the study. All had biopsy-proven carcinoma of the nasopharynx, oropharynx, oral cavity, hypopharynx or larynx, and had received primary radiotherapy with curative intent (> or =50 Gy in 20 fractions) more than 4 months before study entry. More than 75% of both parotid glands were included in the primary radiation field. There was no clinical evidence of recurrent disease. Patients were provided with a 2 month supply of Biotene mouthwash, toothpaste, chewing gum and Oralbalance gel. Response was evaluated 1 and 2 months after study entry using a patient-completed visual analogue scale to assess the severity of xerostomia and its effects on quality of life. For analysis, the scored baseline was subtracted from the later scores to assess change. Patients with an increase of 10 mm from their baseline score on the visual analogue scale were classified as having responded to the treatment intervention, and those with an increase of > or =25 mm from their baseline score were classified as having experienced a major improvement in their symptoms. After 2 months of treatment, 15 patients (54%) reported an improvement in intraoral dryness and 10 of these patients (36%) reported a major improvement. Similar proportions of patients (46% some improvement, 25% major improvement) reported an improvement in their ability to eat normally. Seventeen patients (61%) reported an improvement in oral discomfort, and 12 of these (43%) had a major improvement in their symptoms. The results of this study suggest that the use of Biotene (mouthwash, toothpaste and chewing gum) and Oralbalance gel can improve many of the symptoms of radiation-induced xerostomia. A placebo effect could account for many of the observed improvements in symptoms, and in order to assess the role of these agents in the management of patients with postirradiation xerostomia a randomised phase III study is needed. PMID- 10789962 TI - Oral sequelae of chemotherapy: an important teaching opportunity for oncology health care providers and their patients. AB - The object of this work was to conduct a rapid assessment of a teaching hospital's promotion of optimal oral health among its chemotherapy patients. A pilot study was undertaken, which included focus interviews with oncology clinic staff, a review of the fellowship training curriculum, and unobtrusive observations in the clinic setting. Charts were also audited for oncology patients who were probable chemotherapy candidates. A review of the data offered no evidence that oral health care was routinely addressed in a preventive context prior to the initiation of chemotherapy. Promotion of oral health care will help reduce the risk of oral sequelae of chemotherapy for patients and the subsequent impact of the oral sequelae on patients' chemotherapeutic regimen, thereby improving patients' chances of survival and improving their quality of life. Other teaching hospitals may wish to conduct a similar rapid assessment to determine whether they too could improve patient care and professional education in this area by incorporating pre-chemotherapy oral health evaluation and treatment into routine care for cancer patients. PMID- 10789963 TI - Factors correlated with fatigue in disease-free breast cancer patients: application of the Cancer Fatigue Scale. AB - Fatigue is one of the most frequent symptoms in cancer patients. However, the precise causes of this fatigue are still unknown, and this situation makes it difficult to combat the problem. The present study was conducted to investigate factors correlated with fatigue in disease-free breast cancer patients. A group of 134 randomly selected ambulatory breast cancer patients who had undergone successful surgical treatment participated. They completed the Cancer Fatigue Scale, the Hospital Anxiety and Depression Scale, the Mental Adjustment to Cancer Scale, and an ad hoc questionnaire detailing physical symptoms, social support, and demographic variables at home and returned them by mail the following day. Multiple regression analysis revealed that fatigue was significantly correlated with dyspnea, insufficient sleep, and depression, and that these three variables accounted for a total of 46% of variance in fatigue. Factors concerned with the cancer and treatment, such as disease stage, lymph node metastasis, number of days since operation, past intravenous chemotherapy, radiotherapy, current use of fluoropyrimidine compounds, and current use of tamoxifen citrate were not correlated with fatigue. The results suggest that fatigue in this population is determined by current physical and psychological distress rather than by the cancer itself and prior cancer treatments, and that the management of dyspnea, insomnia, and depression might be important in reducing fatigue in this population. PMID- 10789964 TI - The difficult choice of chemotherapy in patients with unresectable non-small-cell lung cancer. AB - The aims of this study were: to assess the attitude of non-small-cell lung cancer (NSCLC) patients to being treated with chemotherapy, determining whether and how much it differs from that expressed by patients with benign diseases or by healthy people; and to investigate how the information received about the treatment may influence the patients' decisions. A three-item self-assessment questionnaire measuring willingness to be treated with chemotherapy and presented according to three different scenarios (with an optimistic, neutral, and pessimistic physician's presentation) was administered to 104 NSCLC patients, 129 other patients with respiratory diseases (RDP), 140 health care providers (HCP) and 120 students (STU). Guttman's coefficient of reproducibility confirms the hierarchical structure of the three scenarios ranging from an optimistic to a pessimistic view. Relative to the other groups, cancer patients showed: (a) a consistently higher degree of uncertainty about whether to accept or reject chemotherapy; (b) the lowest acceptance rate in the optimistic and neutral scenario and, in contrast, the highest in the pessimistic scenario; (c) the highest percentage of constant answers, independently of the scenario presented, particularly as regards the answers "I don't know" (NSCLC = 25%, RDP = 9%, HCP = 2%, STU = 5%) and "Yes, I accept" (NSCLC = 29%, RDP = 31%, HCP = 19%, STU = 16%). Answer patterns differed markedly between cancer patients, the HCP, and the STU group, and in most cases the difference was statistically significant at a confidence level of 0.001. The differences between NSCLC and RDP patients were less marked, and not always statistically significant. The choice between accepting and rejecting chemotherapy is very difficult for patients with NSCLC, much more so than for healthy people, and it is often independent of the way the information is provided. PMID- 10789965 TI - Prevention of cisplatin-induced delayed emesis: still unsatisfactory. Italian Group for Antiemetic Research. AB - The incidence of and prognostic factors in cisplatin-induced delayed emesis were evaluated in 522 naive cancer patients. They each received an intravenous combination of ondansetron and one of four different doses of dexamethasone for the prevention of acute emesis and a combination of orally administered metoclopramide plus intramuscular dexamethasone for the prevention of delayed emesis. Despite the best prophylaxis for acute and delayed emesis, 37.4%/57.1% of patients experienced delayed vomiting/nausea. The presence of acute vomiting/nausea was the main prognostic factor in delayed vomiting/nausea; therefore, a multifactorial analysis of the results achieved during the delayed phase adjusted for those obtained during the acute phase should be carried out in every study evaluating the efficacy of different antiemetic drugs for delayed emesis. PMID- 10789966 TI - Antiemetic efficacy of combination therapy with granisetron plus prednisolone plus the dopamine D2 antagonist metopimazine during multiple cycles of moderately emetogenic chemotherapy in patients refractory to previous antiemetic therapy. AB - Effective antiemetic treatment of patients who have previously experienced chemotherapy-induced nausea and vomiting is difficult. The aim of this study was to evaluate the antiemetic efficacy of a single intravenous dose of granisetron plus a 3-day oral treatment with prednisolone 25 mg once a day plus metopimazine 30 mg four times a day in patients refractory to previous antiemetic treatment with granisetron or with prednisolone plus metopimazine. The study population was made up of 25 consecutive women with stage I or II breast cancer, who were treated with multiple cycles of adjuvant cyclophosphamide, fluorouracil plus methotrexate or cyclophosphamide, epirubicin plus fluorouracil given i.v. every 3 weeks. Patients received the three-drug combination of antiemetics during a total of 113 cycles of chemotherapy. No emetic episodes were reported in 88.9% cycles on day 1, in 94.7% cycles on days 2 through 5 and in 85.8% cycles on days 1 through 5 after chemotherapy. No nausea was reported in 43.4% cycles on day 1, in 49.6% cycles on days 2 through 5 and in 34.5% cycles on days 1 through 5. Nineteen patients (76.0%) completed the scheduled nine cycles of chemotherapy, 1 being withdrawn because of > or =5 emetic episodes and 5, because they were not satisfied with the antiemetic treatment. The treatment was well tolerated. In conclusion, granisetron plus prednisolone plus metopimazine is a highly effective antiemetic treatment in patients receiving moderately emetogenic chemotherapy refractory to antiemetic therapy with granisetron or prednisolone plus metopimazine. PMID- 10789967 TI - Effect of culture conditions on the formation of struvite by Myxococcus xanthus. AB - The amount of struvite (MgNH4PO4 x 6H2O) produced by Myxococcus xanthus as well as the culture parameter values (pH, total phosphorus, total Kjeldahl nitrogen) were dependent on the culture medium used. Struvite formation started during the exponential phase and the maximum concentration was observed at the beginning of stationary growth phase. The addition of each medium component to the liquid culture influenced the amount of crystal produced. This amount did not depend on the pH increase during the culture period. The moment of the bacterial growth phase, at which each medium component was added, influenced the struvite formation. PMID- 10789968 TI - Copper-catalyzed chlorination and condensation of acetylene and dichloroacetylene. AB - The chlorination and condensation of acetylene at low temperatures is demonstrated using copper chlorides as chlorinated agents coated to model borosilicate surfaces. Experiments with and without both a chlorine source and borosilicate surfaces indicate the absence of gas-phase and gas-surface reactions. Chlorination and condensation occur only in the presence of the copper catalyst. C2 through C8 organic products were observed in the effluent; PCDD/F were only observed from extraction of the borosilicate surfaces. A global reaction model is proposed that is consistent with the observed product distributions. Similar experiments with dichloroacetylene indicate greater reactivity in the absence of the copper catalyst. Reaction is observed in the gas phase and in the presence of borosilicate surfaces at low temperatures. The formation of hexachlorobenzene is only observed in the presence of a copper catalyst. PCDD/F were only observed from extraction of the borosilicate surfaces. A global reaction model is proposed for the formation of hexachlorobenzene from dichloroacetylene. PMID- 10789969 TI - A comparative characterization of dissolved organic matter by means of original aqueous samples and isolated humic substances. AB - The aim of our study is to test the use of less time-consuming spectroscopic methods applied on original water samples in order to obtain information about DOM composition without any sample preparation. These results were directly compared with results from a conventional isolation and characterization procedure of dissolved humic substances (fulvic acids--FA) isolated from the same water sample. FAs were characterized by UV-, fluorescence-, FTIR spectroscopy and elemental composition. UV absorbance and fluorescence behavior of FAs and original water samples follow the same pattern. A lower UV absorbance and a lower humification index (derived from the synchronous fluorescence spectra) of about 15% is typical for water samples compared to the FAs. We computed linear relationships between properties of the original water sample (UV-, synchronous fluorescence spectra) and the isolated FA (IR absorption, C/N ratio). The application of synchronous fluorescence and UV spectroscopy of aqueous samples has been proved to result in similar information about DOM composition as the characterization of isolated humic substances concerning the content of aromatic structures and the degree of humification. PMID- 10789970 TI - Effect of short-term sunlight irradiation on absorbance spectra of chromophoric organic matter dissolved in coastal and riverine water. AB - Samples of riverine and coastal, filtered (filter pore size 0.2 microm) water were exposed to short-term sunlight irradiation which reduced their absorbance in the UV and visible regions. Absorbance losses in coastal chromophoric dissolved organic matter (CDOM) were up to 10-fold smaller than those in riverine CDOM. Accompanying changes of absorbance spectra shapes (increased slope parameter) were, probably, a result of decrease of the mean molecular size of light absorbing organic matter. The potential of coastal CDOM to photodegradation was smaller and was exhausted during the course of a day-long experiment. A distinctive feature of spectral changes after sunlight exposure was a maximum absorbance decrease which appeared at approximately 300 nm in riverine and at approximately 280 nm in coastal water. That selective absorbance loss has been ascribed in both cases to the disappearance of chromophores of terrestrial origin which, in coastal water, had a lower mean molecular size (due to flocculation and/or prior photodegradation) but, nevertheless, retained their molecular properties. PMID- 10789971 TI - The use of PLS algorithms and quantum chemical parameters derived from PM3 hamiltonian in QSPR studies on direct photolysis quantum yields of substituted aromatic halides. AB - The Partial Least-Squares (PLS) method was used to further study Quantitative Structure-Property Relationship (QSPR) for photohydrolysis quantum yields of selected aromatic halides. The cluster of substituted bromobenzenes and iodobenzenes was further grouped into two clusters, substituted bromobenzenes and substituted iodobenzenes, for which QSPRs were obtained, respectively. The QSPRs obtained by PLS are more significant than those obtained in previous studies. The studies showed that only when the compounds belonging to different groups are studies separately, inherent relationships between molecular properties and their structure characteristics can be revealed. It can be generally concluded that the photohydrolysis quantum yields of the substituted aromatic halides are dependent on the overall characters of the molecules, the characters of the carbon-halogen bonds to be broken in the photolysis, and the character of the halogen atoms to be replaced in the photolysis. Electronegativity has great relevance to the photohydrolysis processes of the compounds. Substituted aromatic halides with large average molecular polarizability and molecular weight values tend to have low photolysis quantum yields. PMID- 10789972 TI - Use of diffuse reflectance spectrometry in spot test reactions for quantitative determination of cations in water. AB - Diffuse reflectance spectroscopy can be successfully used for the quantitative determination of small amounts of pollutants like metals. The remission function was found to be linearly proportional to the concentration, when we applied the Kubelka-Munk equation. The color reactions of Cu(II), Co(II), and Ni(II) with dithiooxamide, were realised on filter paper. Reaction between Fe(III) and ammonium thiocyanate was realized on filter paper and gelatine matrix. All measurements were accomplished with a laboratory-constructed reflectometer. We have obtained a calibration curve by plotting the optical density of reflectance A(R) vs log of the mol l(-1) concentration. Limits of detection at the 10(-4) M level were estimated for all the compounds. Linear dynamic range extend over one order of magnitude and shows the potential of device for the quantitative analysis of environmental pollutants. PMID- 10789973 TI - Ecological effects, transport, and fate of mercury: a general review. AB - Mercury at low concentrations represents a major hazard to microorganisms. Inorganic mercury has been reported to produce harmful effects at 5 microg/l in a culture medium. Organomercury compounds can exert the same effect at concentrations 10 times lower than this. The organic forms of mercury are generally more toxic to aquatic organisms and birds than the inorganic forms. Aquatic plants are affected by mercury in water at concentrations of 1 mg/l for inorganic mercury and at much lower concentrations of organic mercury. Aquatic invertebrates widely vary in their susceptibility to mercury. In general, organisms in the larval stage are most sensitive. Methyl mercury in fish is caused by bacterial methylation of inorganic mercury, either in the environment or in bacteria associated with fish gills or gut. In aquatic matrices, mercury toxicity is affected by temperature, salinity, dissolved oxygen and water hardness. A wide variety of physiological, reproductive and biochemical abnormalities have been reported in fish exposed to sublethal concentrations of mercury. Birds fed inorganic mercury show a reduction in food intake and consequent poor growth. Other (more subtle) effects in avian receptors have been reported (i.e., increased enzyme production, decreased cardiovascular function, blood parameter changes, immune response, kidney function and structure, and behavioral changes). The form of retained mercury in birds is more variable and depends on species, target organ and geographical site. With few exceptions, terrestrial plants (woody plants in particular) are generally insensitive to the harmful effects of mercury compounds. PMID- 10789974 TI - Toxic and nutrient element concentrations in soft tissues of zebra and quagga mussels from Lakes Erie and Ontario. AB - Zebra and quagga mussels were collected from Lakes Erie and Ontario in 1997 and the soft mussel tissues were analyzed for Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Se, Sr, V and Zn. No consistent relationships were apparent when comparing element concentrations in soft mussel tissues and mussel type, size range or sampling location. Literature dealing with the absorption of metals by both mussel types is reviewed. PMID- 10789975 TI - Reduction of halogenated hydrocarbons in aqueous media: I. Involvement of sulfur in iron catalysis. AB - Experiments were carried out to investigate the reduction of tri- and tetra chloroethene with iron in aqueous solutions. Results indicate (i) dependence of the dehalogenation capacity of the iron on its sulfur content and (ii) that ethyne was the primary dehalogenated hydrocarbon for both compounds. A reaction mechanism based on in situ formation of ferrous sulfide as the catalyst and its reaction with the halocompounds in the presence of hydrogen has been postulated. PMID- 10789976 TI - Influence of cetane improvers on the air quality in an urban street canyon. AB - The concentration distributions of NOx, PM, HC and CO in an urban street canyon have been estimated using a two-dimensional air quality numerical model based on the k-epsilon turbulent model and the atmospheric convection diffusion equation when various cetane improvers were used in diesel fuels. A wind vortex can be found within the street canyon, and the pollutants emitted from the bottom of the street canyon tend to follow the course of the wind field, moving circularly. The addition of cetane improvers can improve the air quality in a street canyon, all of the pollutants were found to decrease with increasing centane number. PMID- 10789977 TI - An algebraic/graphical tool to compare ecosystems with respect to their pollution V: cluster analysis and Hasse diagrams. AB - In case of large data matrices comparative evaluations of objects/regions with the technique of Hasse diagrams may be troublesome due to a messy system of lines in the graphical representation. Here fuzzy clustering leads to useful simplifications because regions with slightly different pollution pattern are grouped together. However, fuzzy clustering implies to introduce a threshold value for the membership of an object to a cluster and to select the best number of clusters. Therefore many arbitrarities evolve. Within the systematic study presented here we found that some objects are very stable against variations of the threshold value and the number of cluster whereas other objects behaves different. According to their behaviour we investigated a classification of the objects. Formal Concept Analysis shows that in some cases specific pollution pattern imply the membership to one of these classes. For example objects which are characterized by high Pb-, Zn-concentration and moderate S-concentration imply a high stability against variants of the clustering process. Further implications are described in the paper. PMID- 10789978 TI - Experimental determination of the kinetic rate law for the oxidation of perchloroethylene by potassium permanganate. AB - Flushing soils contaminated with trichloroethylene (TCE) and perchloroethylene (PCE) with a permanganate (MnO4-) solution has been shown to reduce the solvent content of the soil. Experiments were performed to quantify the rate at which KMnO4 oxidizes aqueous solutions of PCE over a range of concentrations. In a series of homogeneous reactors, aqueous phase PCE concentrations were monitored over time in nine experimental trials with excess oxidant concentrations ranging from 5 to 30 g/l. Analysis of the data was performed to quantify the oxidation reaction order with respect to PCE and KMnO4 and the reaction rate constant. The reaction between PCE and KMnO4 was determined to be first-order with respect to both PCE and KMnO4 with an overall specific reaction rate coefficient of 2.45+/ 0.65 M(-1) min(-1). PMID- 10789979 TI - A chemostat system for investigating pesticide biodegradation in continuous mixed bacteria cultures originating from surface water. AB - To be able to predict the degradation (rate) of organic chemicals (e.g. pesticides) in the field, knowledge of the environmental conditions that are of influence on the degradation process are of importance. In the present study an experimental system is described which is used to study the degradation of organic pollutants in mixed bacteria cultures originating from surface water With this system the degradation of compounds can be followed for relatively long experimental periods (months). In addition, it is possible to vary different environmental parameters in order to investigate their influences on the degradation of the chemical. These preliminary experiments show that growth and 'composition' of the bacteria culture have comparable patterns in parallel experiments. The first order degradation rate constant for the test compound dichloran, as calculated from these experiments under these circumstances, is about 0.002 h(-1). PMID- 10789981 TI - Kinetics of nitromusk compounds degradation in water by ultraviolet radiation and hydrogen peroxide. AB - The photodegradation of five representative nitromusk compounds in water has been performed in a stirred batch photoreactor with a UV low-pressure immersed mercury lamp, at constant temperature and different doses of hydrogen peroxide. The rate constants have been calculated on the basis of experimental data and a postulated first-order kinetic model. The rate constants, at 298 K and a dose of 1.1746 micromol l(-1) H2O2 ranges from 0.3567 x 10(-3) s(-1) for musk tibetene, to 1.785 x 10(-3) s(-1) for musk ambrette. PMID- 10789980 TI - An investigation of adsorption at the air-water and soil-water interfaces for non micellizing ethylene oxide-propylene oxide surfactants. AB - Adsorption at the air-water interface and soil sorption from aqueous solution have been investigated for a group of ethylene oxide (EO)-propylene oxide (PO) block copolymeric surfactants. The group which have a common structural formula of EOm POn EOm is distinguished by the fact that they have large critical micelle concentration (CMC) values and therefore do not readily form micelles at common environmental concentrations and temperatures. Adsorption at the air-water interface is readily shown to be driven by the size of the hydrophobic PO block. The size of the reduction in surface tension produced by a common concentration of 10(-5) mol dm(-3) linearly increases with the size of the PO block as does the efficiency of adsorption at the air-water interface as measured by pC20--the negative logarithm of the surfactant concentration that produces a reduction in surface tension of 20 mN m(-1). Soil sorption data have also been captured for these compounds and the data are readily fitted to the Freundlich adsorption isotherm. However soil sorption is shown to be inversely related to the molecular mass of the molecules and appears to be related to the size of the hydrophilic EO blocks in the molecule. PMID- 10789982 TI - Solubilization of substituted indole compounds by beta-cyclodextrin in water. AB - The solubilization of four pairs of substituted indole compounds (SICs) by beta cyclodextrin (beta-CD) in water was investigated. The results show that 1,2,3,4 tetrahydrocarbazole and N-methyl-1,2,3,4-tetrahydrocarbazole form 1:1 inclusion complexes with beta-CD, while the other six SICs form 1:2 inclusion complexes, respectively. To each pair of SICs with similar structures, the differences between their solubilization in beta-CD/water solutions has been explained by the difference of their contact area within the beta-CD cavity, the difference of their molecule polarity, or the presence of hydrogen bond between SIC molecule and beta-CD molecule. PMID- 10789983 TI - A novel ortho-dehalogenation reaction of 2-chlorocinnamic acid catalyzed by the pink yeast Rhodotorula rubra Y-1529. AB - In the present study, a resting cells suspension of Rhodotorula rubra Y-1529 was shown to have the capacity to perform an ortho-dehalogenation reaction on 2 chlorocinnamic acid. The results from the biodegradation of U-[14C]benzoic acid, cinnamic acid, 3-chlorocinnamic acid and 4-chlorocinnamic acid suggest that the first step of the ortho-dehalogenation reaction occurred during the oxidation of the unsaturated C3 side chain of 2-chlorocinnamic acid to 2-chlorobenzoic acid. None of the 2-chlorobenzoic acid was found in the biodegradation system, suggesting that this step was a highly regulated step. After the side-chain oxidation reaction, the hydroxylation of the benzene ring was determined to be at the para-position first, followed by the meta-position. The occurrence of 3:4 position ring fission reactions and the production of the final product, CO2, was proven by the biodegradation of U-[14C] benzoic acid. This oxidative dehalogenation reaction catalyzed by R. rubra was found to be regiospecific for 2 chlorocinnamic acid; the chloride ion was probably removed after the ring fission reaction. A pathway of the ortho-dehalogenation reaction of 2-chlorocinnamic acid catalyzed by R. rubra was proposed based on these data. PMID- 10789984 TI - Relationship between electron donor and microorganism on the dechlorination of carbon tetrachloride by an anaerobic enrichment culture. AB - An investigation involving the supplement of different concentrations of substrates and microorganisms was carried out under anaerobic condition to assess the feasibility of bioremediation of carbon tetrachloride (CCl4) with the amendment of low concentrations of auxiliary substrate and microorganisms. The concentrations of substrate and microorganisms ranged from 10 to 100 mg/l and from 3.7 x 10(4) to 3.7 x 10(6) cell/ml, respectively. The biotransformation rate of CCl4 increased progressively with the increase in the concentrations of the substrate and microorganisms. In the low biomass-amended system (3.7 x 10(4) cells/ml), 28-71% and 57-96% of CCl4 removals were exhibited when 10-100 mg/l of acetate or glucose was supplemented, respectively, whereas nearly complete degradation of CCl4 was observed in the heavily inoculated systems (3.7 x 10(6) cells/ml). An addition of electron donor in the low microbial activity batches enhanced greater efficiency in dechlorination than in the high microbial activity batches. The second-order rate constants ranged from 0.0059 to 0.0092 l/mg/day in high biomass input system, while a two- to four-fold increase in rate constant was obtained in the low microbial activity system. This study indicates that biomass was the more important environmental parameter than substrate affecting the fate of CCl4. The addition of auxiliary substrates was effective only in low biomass-amended batches (0.56 mg-VSS/l) and diminished inversely with the increase of microbial concentration. PMID- 10789985 TI - Pathway of anthracene modification under simulated solar radiation. AB - Exposure of polycyclic aromatic hydrocarbons (PAHs) to sunlight results in rapid structural photomodification generally via oxidation reactions. These PAH modification products are in many cases more toxic than their parent compounds. In this study, anthracene (ANT), a rapidly photooxidized PAH, was irradiated with simulated solar radiation (SSR, 100 micromol m(-2) s(-1)) in aqueous solution to examine the photomodification pathway. The photoproducts formed were identified by HPLC. The ANT product profile after 9 h in SSR was very complex, with more than 20 compounds detected. The photoproducts formed were anthraquinones, benzoic acids, benzaldehydes and phenols showing the process to be oxidative in nature. Some of the anthraquinones were themselves subject to photooxidation, and were thus intermediates in the product pathway. The kinetics of ANT photooxidation revealed a pseudo first-order reaction with a half-life of 2 h under the SSR source used. The kinetics of product formation allowed deduction of a probable photomodification pathway. This study indicates that PAH photooxidation products are likely to exist as complex, dynamically changing mixtures in PAH contaminated aquatic environments. PMID- 10789987 TI - Reversal of spontaneous progressive autoimmune encephalomyelitis by myelin basic protein-induced clonal deletion. AB - Autoimmune encephalomyelitis can be initiated spontaneously and developed progressively in TCR transgenic mice specific for myelin basic protein when exposed to non-sterile environment, thus more closely mimicking human multiple sclerosis. By intravenous administration of myelin basic protein, we succeeded in reversing the clinical and pathological signs of progressive spontaneous disease in these mice. Flow cytometry showed that the majority of transgenic T cells in lymph nodes and spleen as well as spinal cords of treated mice were deleted. Dramatically increased numbers of apoptotic cells were found in peripheral immune organs of treated animals. Proliferative responses of single transgenic T cell to autoantigen were significantly decreased in treated mice, indicating that the remaining T cells were anergic. Moreover, production of both Th1 and Th2 cytokines was suppressed. This study is the first demonstration of reversal of progressive, spontaneous autoimmune disease of the central nervous system, and provides direct evidence that apoptosis-induced clonal deletion, along with anergy of remaining cells, but not Th2 switch, play a major part in the reversal of this disease by intravenous administration of autoantigen. PMID- 10789986 TI - Guest dechlorination and covalent capture following photoexcitation of inclusion complexes in water. AB - Photoexcitation of complexes between cyclophane 1 and 1- or 2-chloronaphthalene in aqueous solution leads to rapid dechlorination of the guest, a reaction driven by electron transfer from host to excited guest. The main photoproducts contain a naphthyl group covalently attached to the host framework. The results may lead to new approaches for remediating water contaminated with chlorinated aromatic compounds. PMID- 10789988 TI - p53 autoantibodies in patients with autoimmune diseases: a quantitative approach. AB - With few exceptions, autoantibodies directed against the gene product of the tumor suppressor gene p53 are only detected in cancer patients. From 73 patients with various autoimmune diseases, we obtained 17 sera with elevated autoantibodies against the p53 protein comprising patients with SLE, Graves' disease, and immune vasculitis including Wegener's granulomatosis. The overall prevalence (23%) of p53 autoantibodies was comparable to that in various cancers; differences, however, were obvious with respect to the magnitude of antibody levels. Only 5% of seropositive colorectal cancer patients had levels within the critical range (150-180 U/ml) but nearly half (41%) of seropositive autoimmune disease patients were that low. None of the autoimmune disease patients exceeded 300 U/ml serum compared to more than 60% of seropositive colorectal cancer patients with higher levels. This remarkable difference in magnitude underlines the necessity of quantification of p53 autoantibodies over a mere qualitative determination. Patients with autoimmune diseases face an increased risk for malignancies. It still remains to be established whether p53 seropositivity in autoimmune diseases adds to the rare exceptions of p53AAb in non-malignant diseases or is indicative for a yet occult cancer. PMID- 10789989 TI - Immunoregulation of encephalitogenic MBP-NAc1-11-reactive T cells by CD4+ TCR specific T cells involves IL-4, IL-10 and IFN-gamma. AB - The generation of TCR transgenic (Tg) mice expressing a BV8S2 (Vbeta8 subfamily 2) chain specific for the encephalitogenic NAc1-11 region of MBP provides a unique system for evaluating the mechanisms involved in anti-TCR immunoregulation of EAE. In a previous study, we showed that vaccination with BV8S2 protein induced specific T cells that inhibited proliferation responses and encephalitogenic activity of MBP-reactive T cells in vitro, and resulted in a skewed production of Th2 cytokines by the MBP-reactive T cells. These data suggested that regulation of the encephalitogenic T cells was mediated by inhibitory cytokines rather than through a deletional mechanism. In the current study, we have employed the BV8S2 Tg mouse model to address the issue of which cytokines produced by anti-TCR-reactive T cells can regulate the function of encephalitogenic Th1 cells. Utilizing neutralizing anti-cytokine antibodies to reverse inhibitory effects of supernatants from BV8S2-specific T cells, we found that IL-4, IL-10, and to a lesser extent, IFN-gamma and TGF-beta, were the major regulatory cytokines responsible for inhibiting encephalitogenic activity, proliferation, and IFN-gamma secretion of MBP-NAc1-11-reactive Th1 cells. These results indicate that cytokine regulation is the major mechanism through which TCR specific CD4+ T cells regulate encephalitogenic and potentially other bystander Th1 cells. PMID- 10789990 TI - Disorganization of thymic medulla precedes evolution towards diabetes in female NOD mice. AB - The thymic medulla is a complex microenvironment which plays a crucial role in central tolerance induction. Using a quantitative histological analysis of non obese diabetic (NOD) mice, we show that the medulla undergoes several structural modifications during the course of the disease in NOD mice. Indeed, the majority of 70-day-old NOD mice show a scattering of medullary epithelial cells in the cortex which is associated with a reduction in the size of the medulla in heavily disorganized thymuses. The severity of this phenotype is shown to correlate with the subsequent appearance of diabetes in older female NOD mice. This trait is mainly controlled by non-major histocompatibility complex NOD genes since C57BL/6 H-2g(7) congenic mice have a normal medulla. It persists in conditions where effector lymphocytes that lead to diabetes are inhibited in periphery. These results suggest that primary alterations of the thymic stroma might play a role in the progression towards diabetes in NOD mice. PMID- 10789991 TI - Higher incidence of autoantibodies in X-linked chronic granulomatous disease carriers: random X-chromosome inactivation may be related to autoimmunity. AB - The presence of autoantibodies and autoimmune diseases was tested in all available members of five families with at least one member affected with X linked chronic granulomatous disease. Patients and carriers relatives possess autoantibodies more frequently than non-carriers relatives (95% vs 10%, p < 1.0 x 10(-5), Fisher test). Further, a survey of the literature revealed that in X linked immunodeficiencies with X-chromosome random inactivation, clear features of autoimmunity are observed, not found in those with non-random inactivation. It appears then as if random inactivation of the X-chromosome in these pathologies, may favor the expression of an autoimmune phenotype in patients and carriers. PMID- 10789992 TI - A novel mutation in the FcgammaRIIIA gene (CD16) results in active natural killer cells lacking CD16. AB - We report a novel mutation in FcgammaRIIIA (the transmembrane-form CD16) on natural killer (NK) cells in a patient with polyneuropathy. She had no history of recurrent infections. Her NK cells expressed no detectable CD16; however, her NK cytotoxic activity was normal, suggesting that CD16 expression and cytotoxic activity are independent of one another. Molecular analysis revealed a deletion of a single adenine base in exon 4 of CD16 at nucleotide 550. This deletion generates a STOP codon in an extra-cellular domain of the FcgammaRIIIA gene, thereby truncating the CD16 molecule. The patient's NK cells were not recognized by the anti-CD16 monoclonal antibodies 3G8 and Leu11c. Whether the development of her polyneuropathy is associated with this novel mutation is unclear. PMID- 10789993 TI - Experimental autoimmune thyroiditis induced by thyroglobulin-pulsed dendritic cells. AB - Dendritic cells (DCs), which are the most effective professional antigen presenting cells (APCs), initiate and regulate immune responses. In this report, we examine the role of DCs in the induction of autoimmune thyroiditis. Experimental autoimmune thyroiditis (EAT) induced by immunization with thyroglobulin (Tg) plus adjuvant is considered to be an animal model of autoimmune thyroiditis, and is categorized as a T cell-mediated autoimmune disease. To examine the contribution of DCs to EAT, naive DCs were purified from high responder B10BR mice and pulsed with various concentrations of porcine Tg (pTg). These pTg-pulsed DCs were transferred without adjuvant to syngenic BIOBR mice to induce EAT. Mice that had received pTg-pulsed DCs showed thyroiditis, and the degree of thyroiditis induced was positively correlated to the amounts of pTg used for the incubation (pulsing) of DCs. The severity of thyroiditis was also correlated to the amounts of anti-pTg IgG2a antibodies and IFN-gamma in the recipient sera, but not to IL-4 or IL-10, indicating that Th1 cells are mainly activated by pTg-pulsed DCs and attributable to the pathogenesis of EAT. PMID- 10789994 TI - Pathogenesis and therapeutic concepts of organ-specific autoimmune diseases: report of an interdisciplinary workshop in Wurzburg, April 29-30, 1999. PMID- 10789995 TI - Psychosocial treatments in schizophrenia: a review of the past 20 years. AB - Pharmacotherapy can improve some of the symptoms of schizophrenia but has limited effect on the social impairments that characterize the disorder and limit functioning and quality of life. Through computerized literature searches and bibliographies of published reports we identified peer reviewed studies of group, family, and individual therapy with schizophrenia and schizoaffective disorder patients. We identified 70 studies: 26 on group therapy, 18 on family therapy, and 11 on individual therapy. Additionally, treatment models were compared in 4 studies and combined in 11 others. Controls were included in 61 and all studies included medication. Benefits in symptoms as well as social and vocational functioning were associated with psychosocial treatments. Family therapy demonstrated the most promising findings and traditional social skills treatment yielded the least robust results. Adjunctive psychosocial treatments augment the benefits of pharmacotherapy and enhance functioning in psychotic disorders. Although these positive results have led to increased enthusiasm about psychosocial treatments for schizophrenia, questions remain about comparative benefits of specific treatment methods and additional benefits of multiple treatments. PMID- 10789996 TI - Long-term follow-up of patients with a diagnosis of paranoid state and hospitalized, 1913 to 1940. AB - From a sample of 239 patients diagnosed paranoid state and hospitalized between 1913 and 1940 at the Phipps Clinic, we particularly studied a group of 60 patients without previous hospitalizations, consisting of 57 patients with follow ups of 5 or more years, and 3 patients who killed themselves (the ultimate follow up) less than 1 year after discharge. These 60 patients had been retrospectively diagnosed with delusional disorders by DSM-IV criteria. On follow-up, 27% were rated recovered, whereas 52% were rated unimproved. Long-term follow-up was correlated with discharge status. Poor follow-up was significantly correlated with seclusive personality, poor premorbid history, onset 6 months or more before admission, gradual onset, lack of insight, single marital status, and lack of precipitating events. A prognostic scale constructed from the first four of these variables was predictive of long-term outcome. More recent, better treatment results have been contrasted with these findings from an earlier non-drug treatment era. PMID- 10789997 TI - Benchmarking treatment of schizophrenia: a comparison of service delivery by the national government and by state and local providers. AB - This study compared treatment of schizophrenia in two types of organization: a national, government-operated health care system, the Department of Veterans Affairs (VA), and in hospitals and clinics operated by state and local providers. Between 1994 and 1996, 746 male patients with a clinical diagnosis of schizophrenia residing in two states in the Southeast and Midwest were surveyed: 192 VA inpatients were compared with 96 non-VA inpatients, and 274 VA outpatients were compared with 184 non-VA outpatients. VA patients were older and had higher incomes than non-VA patients but did not differ significantly on measures of clinical status, satisfaction with providers, or community adjustment. VA outpatients were more likely to have been hospitalized during the previous year than non-VA outpatients and were less likely to have received services from a day hospital, from a case manager or social worker, or to have received crisis intervention services. On 5 of 26 Schizophrenic Patient Outcomes Research Team treatment recommendations, a smaller proportion of VA than non-VA patients adhered to standards. Four of these reflected reduced access among VA patients to psychosocial services such as work therapy, job training, or case management services. Cross-sectional surveys can be used to compare quality of care across service systems. VA care was associated with similar satisfaction and clinical outcomes but greater reliance on hospital treatment and less use of community based psychosocial services. PMID- 10789998 TI - Psychotic features in chronic posttraumatic stress disorder and schizophrenia: comparative severity. AB - Psychotic features are frequent in combat veterans with chronic posttraumatic stress disorder (PTSD), may correlate with severity of PTSD symptoms, and may reflect a distinct subtype of the disorder. These psychotic features include auditory and visual hallucinations and delusional thinking that is usually paranoid in nature. Psychotic features may be under-recognized in chronic PTSD because patients are reluctant to report these symptoms and because they may not have overt changes in affect or bizarre delusions characteristic of other psychoses, e.g., schizophrenia. To further assess these phenomena, we compared clinical ratings on the Positive and Negative Syndrome Scale (PANSS) and other assessments, including the Clinical Global Impression Scale and the Structured Clinical Interview with Psychotic Screen, in veterans meeting DSM-IV criteria for chronic PTSD with well-defined comorbid psychotic features (N = 40) or chronic schizophrenia (N = 40). The patients with schizophrenia had modestly higher composite PANSS scores and positive symptom scores although average scores in both groups were moderate to severe in intensity. Negative symptom and general psychopathology subscale scores were comparable in both groups. Regarding specific positive symptoms, hallucinations were comparable between groups in severity; however, schizophrenia patients had slightly more intense delusions and conceptual disorganization. These data further validate the occurrence of positive as well as negative symptoms of psychosis in chronic PTSD in a range of severity that may approach that of patients with schizophrenia. Although meeting DSM-IV criteria for two different major psychiatric disorders, these two patient populations were remarkably similar with respect to not only positive but also negative symptoms. PMID- 10789999 TI - Ethnocultural differences in sleep complaints among adolescents. AB - Data from a school survey (N = 5423) were used to examine differences in sleep complaints among adolescents from nine ethnocultural groups. Compared with Anglo youths, Chinese American youths were at significantly lower risk for insomnia, whereas Mexican American youths had an elevated risk, adjusting for the effects of age, gender, and socioeconomic status. Adjusting for the same three covariates, African, Mexican and Central American youths were at elevated risk for hypersomnia. No group was at lower risk for hypersomnia than the Anglos. The results suggest minority status may affect risk for sleep problems but that the direction and the magnitude of the effects vary depending on the outcome examined, severity of the problem, and the ethnic group studied. Given the paucity of data on culture and risk for disturbed sleep among adolescents, more research is needed on the role of ethnocultural background on sleep. PMID- 10790000 TI - Effect of stimulant medication on driving performance of young adults with attention-deficit hyperactivity disorder: a preliminary double-blind placebo controlled trial. AB - Driving performance of adult males with attention-deficit hyperactivity disorder (ADHD) was compared with matched controls in a double-blind (Ritalin vs. placebo) cross-over design, using a high-fidelity driving simulator. Seven ADHD and six non-ADHD drivers (mean age 22) were screened to rule out comorbidity and assess for ADHD, and then admitted to the General Clinical Research Center to control diet and sleep before testing. At 0800 and 1530, subjects consumed either a placebo or Ritalin pill in a counter-balanced manner, and at 0930 and 1700, subjects drove the simulator. After both drives, subjects rated their driving performance. Compared with non-ADHD subjects, ADHD subjects had more career driving accidents (p < .04) and motor vehicle violations (p = .059), drove worse on the simulator under placebo condition (p < .05), demonstrated significant improvement under the Ritalin condition (p < .05), rated themselves as driving poorer during the placebo condition (p = .05), and tended to perceive their driving to be better during the Ritalin condition (p = .07). This would suggest that individuals with ADHD should have the therapeutic benefit of a stimulant medication when operating a vehicle. PMID- 10790001 TI - Insight and suicidality in schizophrenia: a replication study. PMID- 10790002 TI - Psychometric properties of the Japanese version of the Calgary Depression Scale for Schizophrenics. PMID- 10790003 TI - Stigmatizing perception of mental illness by Japanese students: comparison of different psychiatric disorders. PMID- 10790004 TI - Increased risk for dementia in elderly psychiatric inpatients with late-onset major depression. PMID- 10790006 TI - Edge diffraction of creeping rays AB - Explicit formulas are derived for waves modeled by the scalar two-dimensional Helmholtz equation for the field that is diffracted when surface creeping rays encounter an infinitely sharp edge. Both Neumann and Dirichlet boundary conditions are analyzed, and the diffracted field is found to be an order to magnitude smaller in the latter case. PMID- 10790005 TI - A new formalism for time-dependent wave scattering from a bounded obstacle AB - A time-dependent three-dimensional acoustic scattering problem is considered. An incoming wave packet is scattered by a bounded, simply connected obstacle with locally Lipschitz boundary. The obstacle is assumed to have a constant boundary acoustic impedance. The limit cases of acoustically soft and acoustically hard obstacles are considered. The scattered acoustic field is the solution of an exterior problem for the wave equation. A new numerical method to compute the scattered acoustic field is proposed. This numerical method obtains the time dependent scattered field as a superposition of time-harmonic acoustic waves and computes the time-harmonic acoustic waves by a new "operator expansion method." That is, the time-harmonic acoustic waves are solutions of an exterior boundary value problem for the Helmholtz equation. The method used to compute the time harmonic waves improves on the method proposed by Misici, Pacelli, and Zirilli [J. Acoust. Soc. Am. 103, 106-113 (1998)] and is based on a "perturbative series" of the type of the one proposed in the operator expansion method by Milder [J. Acoust. Soc. Am. 89, 529-541 (1991)]. Computationally, the method is highly parallelizable with respect to time and space variables. Some numerical experiments on test problems obtained with a parallel implementation of the numerical method proposed are shown and discussed from the numerical and the physical point of view. The website: http://www.econ.unian.it/recchioni/w1 shows four animations relative to the numerical experiments. PMID- 10790007 TI - Enhancement and inhibition of acoustic radiation in spherical cavities AB - Acoustic radiation from a source located inside layered cavities is studied using the transfer matrix method. It is shown that the radiation can be either enhanced or inhibited, depending on the characteristics and the material composition of the cavities. The analysis predicts well-defined spectral peaks and nodes in some cavities. It is found that significant enhancement and inhibition are possible by varying acoustic parameters. The results also show that the strength of the radiating source can be significantly modified due to the presence of layered structures. The link to the similar phenomena in the optical cavities is discussed. PMID- 10790008 TI - In-plane elastic property characterization in composite plates AB - This article presents a method to deduce the in-plane elastic properties of multilayered composite plates. Drawing on a synthetic-aperture technique developed for the elucidation of materials properties in air-coupled ultrasonics, this new method exploits the high elastic anisotropy of composite materials to permit an accurate measurement of directional in-plane stiffness. It is found that comparisons of experimental measurements with plate stiffnesses calculated on the basis of lamination theory agree to within several percent for uniaxial and biaxial laminates and to within 10 percent for quasi-isotropic laminates. It is further shown that the method is largely insensitive to transducer deployment angle within a range related to the transducer beamwidth. PMID- 10790009 TI - Acoustic wave motion along a narrow cylindrical duct in the presence of an axial mean flow and temperature gradient AB - A numerical solution is presented to the problem of nonisentropic acoustic wave motion in a circular capillary tube in the presence of both axial mean flow and a background axial temperature gradient. The effects of the radial components of the acoustic velocity are included in the analysis. The main application area is in the study of the acoustic effects of catalytic converters. The solution makes use of a series expansion and is valid for small relative changes in the background temperature, which are typical of this application area. Various solutions to the problem have been obtained previously, using different simplifications to the complete problem which is considered here. It is shown that each of the simplifications results in errors for the predicted attenuation of at least 20 dB/m, using conditions typical for catalytic converters. In particular, the isentropic assumption is shown to be invalid. PMID- 10790010 TI - Acoustic hybrid junction in a rectangular waveguide AB - Acoustic scattering from a hybrid junction in a rectangular waveguide is studied. The Fourier transform and mode matching is used to represent the scattered wave and the simultaneous equations for the modal coefficients are formulated. A residue calculus is utilized to obtain a solution to the simultaneous equations in fast-converging series. Numerical computations are performed to show the behavior of transmission in terms of junction geometry and operating frequency. The utility of acoustic hybrid junction is discussed in terms of its decoupling characteristics. PMID- 10790011 TI - A study of the influence of mean flow on the acoustic performance of Herschel Quincke tubes AB - In this paper, a simple flow model is used in order to assess the influence of mean flow and dissipation on the acoustic performance of the classical two-duct Herschel-Quincke tube. First, a transfer matrix is obtained for the system, which depends on the values of the Mach number in the two branches. These Mach numbers are then estimated separately by means of an incompressible flow calculation. Finally, both calculations are used to study the way in which mean flow affects the position and value of the characteristic attenuation and resonances of the system. The results indicate the nontrivial character of the influence observed. PMID- 10790012 TI - Physical source realization of complex source pulsed beams AB - Complex source pulsed beams (CSPB) are exact wave-packet solutions of the time dependent wave equation that are modeled mathematically in terms of radiation from a pulsed point source located at a complex space-time coordinate. In the present paper, the physical source realization of the CSPB is explored. This is done in the framework of the acoustic field, as a concrete physical example, but a similar analysis can be applied for electromagnetic CSPB. The physical realization of the CSPB is addressed by deriving exact expressions for the acoustic source distribution in the real coordinate space that generates the CSPB, and by exploring the power and energy flux near these sources. The exact source distribution is of finite support. Special emphasis is placed on deriving simplified source functions and parametrization for the special case where the CSPB are well collimated. PMID- 10790013 TI - Calculation of surface wave motions due to a subsurface point force: an application of elastodynamic reciprocity AB - The elastodynamic reciprocity theorem for two time-harmonic elastodynamic states of the same body is used to determine the surface wave motions generated by a subsurface point load of arbitrary direction in an isotropic, homogeneous, elastic half-space. The actual surface wave motions due to the point load expressed in a suitably general form represent one of the states, while the other one is an appropriately selected auxiliary solution consisting of incoming and outgoing surface waves. A direct application of the reciprocity theorem yields the desired information on the generated surface wave motion. PMID- 10790014 TI - Wave-number-based assessment of the doubly asymptotic approximation. I. Frequency domain wet surface impedance AB - The doubly asymptotic approximation (DAA) is a canonical relationship for the interaction between surface normal velocity and pressure. Its validity for a slender hemicapped cylinder is examined by formulating a frequency domain version of DAA using the global basis functions employed in the wave-number-based formulation of the surface variational principle [K. Wu and J. H. Ginsberg, ASME J. Vib. Acoust. 120, 392-400 (1998)]. The wet surface impedance matrix, which relates the spectral representation of normal velocity to a corresponding representation of pressure, is obtained according to a second-order version of DAA and according to the surface variational principle. Comparison and interpretation of the results reveals that DAA fails to account for highlights associated with transition from supersonic to subsonic surface waves as the surface wavelength decreases with frequency held constant. PMID- 10790015 TI - Wave-number-based assessment of the doubly asymptotic approximation. II. Frequency and time domain response AB - In part I, the surface variational principle (SVP) and the doubly asymptotic approximation (DAA) were used to generate alternative descriptions of the frequency domain wet surface impedance. These are used here to obtain alternative descriptions of the displacement field in slender hemicapped cylindrical shells. This field is represented as a decomposition into a two-dimensional wave-number space. The responses that are obtained are frequency domain transfer functions for a point force at the midplane, evaluated for a discrete set of frequencies. Solutions obtained from first- and second-order versions of DAA are compared to the SVP result. Then temporal responses are obtained by using an inverse fast Fourier transform to evaluate the convolution of the transfer functions with a prototypical temporal excitation. The results reveal that DAA and SVP differ most in the vicinity of fluid-loaded resonances. The largest disagreement is encountered for the axisymmetric portion of the response. It is shown that DAA often greatly underpredicts peak displacements and accelerations, but a few cases lead to overprediction. PMID- 10790016 TI - Experimental investigations of nonlinear acoustic phenomena in polycrystalline zinc AB - The results of experimental and theoretical investigations of nonlinear acoustic phenomena (nonlinear losses, shift of resonance frequency, generation of the third harmonic, and nonlinear sound-by-sound damping) in polycrystalline zinc nonannealed and annealed resonating rods are presented. The measurements were carried out in the 10(-7)-10(-5) strain range at a frequency of about 3 kHz; the frequency of the weak ultrasonic pulse was about 270 kHz. The experimentally observed phenomena are described in frames of phenomenological equations of state containing elastic hysteresis and dissipative nonlinearity. The nonlinear acoustic parameters of these equations are determined by comparison between theoretical dependencies and experimental results. The influence of structural changes in zinc due to annealing on the nonlinear acoustic phenomena is shown. PMID- 10790017 TI - A method for estimating time-dependent acoustic cross-sections of bubbles and bubble clouds prior to the steady state AB - Models for the acoustic cross-sections of gas bubbles undergoing steady-state pulsation in liquid have existed for some time. This article presents a theoretical scheme for estimating the cross-sections of single bubbles, and bubble clouds, from the start of insonation onward. In this period the presence of transients can significantly alter the cross-section from the steady-state value. The model combines numerical solutions of the Herring-Keller model with appropriate damping values to calculate the extinction cross-section of a bubble as a function of time in response to a continuous harmonic sound field (it is also shown how the model can be adapted to estimate the time-dependent scatter cross-section). The model is then extended to determine the extinction cross section area of multiple bubbles of varying population distributions assuming no bubble-bubble interactions. The results have shown that the time taken to reach steady state is dependent on the closeness of the bubble to resonance, and on the driving pressure amplitude. In the response of the population as a whole, the time to reach steady state tends to decrease with increasing values of the driving pressure amplitude; and with the increasing values of the ratio of the numbers of bubbles having radii much larger than resonance to the number of resonant bubbles. The implications of these findings for the use of acoustic pulses are explored. PMID- 10790018 TI - Backscattering enhancements associated with subsonic Rayleigh waves on polymer spheres in water: observation and modeling for acrylic spheres AB - Unlike most common solids, "plastic" polymer solids typically have shear and Rayleigh wave phase velocities less than the speed of sound in water. Subsonic Rayleigh waves on smooth objects in water are not classified as leakey waves and it is necessary to reexamine backscattering mechanisms. Also the intrinsic material dissipation of the Rayleigh wave can be significant. Backscattering by acrylic or polymethlmethacrylate (PMMA) spheres in water is analyzed and measured in the region ka = 1.5-7 and it is found that prominent low-lying resonance peaks of the form function f exist. The peaks can be modeled with quantitative ray theory as the result of coupling of subsonic Rayleigh waves with sound through acoustic tunneling. The most prominent maximum of f=5.63 occurs at ka = 1.73 and is associated with the quadrupole (or n=2) partial wave. In addition to explaining the scattering, the target strength is found to be sufficiently large that such spheres may be useful for passive low frequency targets. PMID- 10790019 TI - A mapping approach for handling sloping interfaces AB - A mapping approach for handling sloping interfaces in parabolic equation solutions is developed and tested. At each range, the medium is rigidly translated vertically so that a sloping interface becomes horizontal. To simplify the approach, the slope is assumed to be small and the extra terms that arise in the wave equation under the mapping are neglected. The effects of these terms can be approximately accounted for by applying a leading-order correction to the phase. The mapping introduces variations in topography, which are relatively easy to handle for the case of a pressure-release boundary condition. The accuracy of the approach is demonstrated for problems involving fluid sediments. The approach should also be accurate for problems involving elastic sediments and should be useful for solving three-dimensional problems involving variable topography. PMID- 10790020 TI - Environmentally adaptive wedge modes AB - The notion of environmentally adaptive, curvilinear, uncoupled normal modes is developed by writing the Helmholtz equation in curvilinear coordinates. The depth dependent vertical scaling function provides some coordinate freedom missing from the Cartesian formulation. This freedom permits adaptation of the normal mode approach to variable depth and variable sound-speed problems for environments that change slowly in range. A W.K.B. approximation to the eigenfunctions shows modal decoupling can be realized when the vertical scaling function is chosen equal to the reciprocal of the depth-dependent vertical wave number of the lowest mode. Numerical simulations demonstrate that a requirement for constant inter modal ratios of the vertical wave number is valid throughout several wedges with different bottom boundary conditions. The horizontal refraction of such modes agrees closely with analytic solutions. For a wide-angle wedge, the acoustic field agrees qualitatively well with the benchmark solution of Dean and Buckingham [J. Acoust. Soc. Am. 93, 1319-1328 (1993)]. PMID- 10790021 TI - Three-dimensional density spectra of sandy sediments AB - Power spectra of density variability for sandy sediments offshore Panama City, Florida, are estimated and modeled using digital x-ray computed tomography images of sediment structure. Spectral analysis reveals that while shell pieces and mud mixtures are the main contributors to density variability at large scales, intrinsic density variability associated with sand grain contacts dominates at small scales. The power spectrum of sandy sediments is modeled by an analytic form that consists of two power-law components, one associated with the shell and mud contributions and the other with the intrinsic density variability of sand. The dominant term has a much higher power-law exponent than previously reported. Implications for the scattering of high-frequency sound in sandy sediments are discussed. PMID- 10790022 TI - An energy-conserving spectral solution AB - An energy-conserving spectral solution is derived and tested. A range-dependent medium is approximated by a sequence of range-independent regions. In each region, the acoustic field is represented in terms of the horizontal wave-number spectrum. A condition corresponding to energy conservation is derived for the vertical interfaces between regions. The accuracy of the approach is demonstrated for problems involving sloping ocean bottoms. The energy-conserving spectral solution is less efficient than the energy-conserving parabolic equation solution, but might be suitable for generalization to problems involving elastic bottoms. PMID- 10790023 TI - Hybrid geoacoustic inversion of broadband Mediterranean Sea data AB - This paper describes an acoustic experiment (PROSIM'97) carried out to investigate inversion for seabed properties at a site off the west coast of Italy where previous acoustic and geophysical studies have been performed. Acoustic fields were measured at a vertical hydrophone array due to a swept-frequency source towed over weakly range-dependent bathymetry. Based on the known geology, the seabed is modeled as a sediment layer overlying a semi-infinite basement with unknown model parameters consisting of the sediment thickness, sediment and basement sound speeds, source range and depth, water depth at the source and array, and array tilt. A hybrid inversion algorithm is applied to determine the model values that minimize the mismatch with the measured acoustic fields. Multiple data sets are analyzed to examine the consistency of the inversion results. It is found that the low sound speed of the sediment layer, together with a large uncertainty in bathymetry, leads to strong correlations between the water depths and sediment thickness. This precludes reliable estimation of these parameters individually; however, the total depth to the basement can be estimated reliably. In addition, the basement speed and geometric parameters are estimated consistently, and all parameters compare favorably with the geophysical ground-truth information and with previous inversion results. PMID- 10790024 TI - Estimation of coherent detection performance for spread scattering in reverberation-noise mixtures AB - Narrow-band matched filter processing gain is estimated for medium- and high frequency active sonars for which the random backscattering processes are assumed to be wide sense stationary in time frequency and uncorrelated in delay-Doppler wide sense stationary and uncorrelated scattering [WSSUS conditions]. Echo and reverberation processes that are WSSUS are described by two-dimensional scattering functions defined in the delay-Doppler plane. The average receiver responses are estimated from the convolution of the appropriate scattering function with the waveform ambiguity function. Estimates of matched filter processing gain are derived for continuous wave (CW) linear frequency modulation (LFM), and discrete frequency shift keyed (FSK) (hop code) waveforms reflected from point and uniform delay spread scatterers masked by reverberation. These bound matched filter performance for a particular waveform and interference distribution since most delay spread scattering falls somewhere between these extremes. The scattering and ambiguity functions are modeled by bounded constant amplitude functions in delay-Doppler that permit the convolutions to be approximated by overlapping area calculations. The results are presented in tabular form as simple formulas that are functions of the reverberation, noise, and waveform parameters. The estimates are shown to be consistent with processing gain measurements made from multiple realizations of synthesized and in-water data. PMID- 10790025 TI - Measurements of nanometer scale interface diffusion between tungsten and niobium thin films using high frequency laser based ultrasound AB - This article presents the use of a quantitative analysis technique to describe time-resolved acoustic spectroscopy (high frequency laser based ultrasound) measurements of atomic diffusion on nanometer length scales occurring at the interface between sputter-deposited tungsten and niobium films. The extent of diffusion at the tungsten-niobium interface is determined by comparing experimental, simulated, and theoretical transfer functions between acoustic arrivals. The experimental and simulated transfer functions use the spectral content of successive reflected acoustic waves and the theoretical transfer function is based on the transfer matrix of an equivalent stratified interface region. This combination of theoretical, simulated, and experimental analyses makes it possible to separate signals with distinct differences between the as deposited interface and those interfaces diffused to an experimentally determined 0.8-nm and 1.4-nm extent. Comparison of predicted and measured diffusion depths for this diffusion couple indicates that bulk diffusivities are not appropriate for describing nanometer scale interface diffusion. PMID- 10790026 TI - Rigid piston approximation for computing the transfer function and angular response of a fiber-optic hydrophone. AB - The transfer function of a fiber-optic hydrophone (FOH) is computed for various fiber core radii. The hydrophone is modeled as a rigid disk, with plane waves impinging at normal or oblique incidence. The total sound field is written as the sum of the incident field and the field diffracted from the hydrophone. The diffracted field is approximated by the field generated by a vibrating planar piston in an infinite rigid baffle. For normal incidence and a pointlike fiber core, an analytical solution is presented. For finite fiber core radii, and for oblique incidence, the transfer functions are computed numerically. The calculated transfer functions exhibit an oscillatory frequency dependency that is most pronounced for small fiber cores. The solution for a core radius of 2.5 microm can be very well approximated by the analytical solution for a pointlike core at frequencies of up to 30 MHz. The results for normal incidence can be directly employed to deconvolute ultrasonic pressure signals measured with an FOH. From the transfer functions for oblique incidence, the angular response of the hydrophone is calculated. The angular response obtained here differs significantly from the model commonly used for piezoelectric hydrophones. The effective hydrophone radius derived from the angular response shows a strong frequency dependency. For low frequencies, it is found to be larger than the outer fiber radius, whereas it generally lies between the outer radius and the fiber core radius for frequencies above 10 MHz. PMID- 10790027 TI - A perturbation technique for the prediction of the displacement of a line-driven plate with discontinuities AB - A novel technique is presented for obtaining approximate analytic expressions for an inhomogeneous line-driven plate. The equation of motion for the inhomogeneous plate is transformed, and the transform of the total displacement is written as a sum of the solution for a homogeneous line-driven plate plus a term due to the inhomogeneity. The result is an integral equation for the transform of the inhomogeneous contribution. This expression may in general be solved numerically. However, by introducing a small parameter into the problem, it may be solved approximately using perturbation techniques. This series may not be convergent, but its convergence may be improved using Pade approximation. Results are presented for the case of a single mass discontinuity, and a distribution of mass discontinuities. PMID- 10790028 TI - Estimation of fuzzy structure parameters for continuous junctions AB - The fuzzy structure theory introduced 15 years ago is designed to predict the frequency response functions of structures and structural acoustic systems with structural complexity, in the low- and medium-frequency ranges. This paper constitutes a first validation of the fuzzy structure theory for continuous junctions between the master structure and the fuzzy substructures. In addition, a method to estimate the fuzzy structure parameters introduced in the fuzzy structure theory is presented and it is validated for the case of continuous junctions. This validation obtained by numerical simulations opens the field of experimental identifications. PMID- 10790029 TI - On acoustic and structural modal cross-couplings in plate-cavity systems AB - Modal cross-couplings are sometimes neglected in the prediction of sound field and structural responses of vibroacoustic systems where an enclosed sound field is coupled to a vibrating boundary structure. In such systems, there are two types of modal cross-couplings and they are commonly referred to as acoustic modal cross-coupling (ACC) and structural modal cross-coupling (SCC). The prediction errors generated from neglecting either of these cross-couplings are dependent not only on the modal properties of the vibroacoustic system (e.g., modal densities, dampings, etc.), but also on whether the sound field or the structure is directly driven. However, the physical mechanisms and characteristics of both cross-couplings are not well understood and, consequently, the conditions when ACC or SCC has a significant contribution to the system responses become unknown. This paper presents a mathematical description which allows the two types of modal cross-couplings to be studied independently. This description is then used to obtain the physical mechanisms and features of both cross-couplings. The effects of each type of cross-couplings on the system responses are then investigated and the general conditions under which these modal cross-couplings may be ignored are underlined. PMID- 10790030 TI - Experimental results for multichannel feedforward ANC with noninvasive system identification AB - This paper presents experimental validation of a class of algorithms designed to enable active noise control (ANC) to function in environments when transfer functions change significantly over time. The experimental results presented are for broadband, local quieting in a diffuse field using a multichannel ANC system. The reverberant enclosure is an ordinary room, measuring approximately 1.4 x 2.4 x 2.4 m3 and containing a seated occupant, with six microphones defining the quiet zone near the occupant's ears. The control system uses a single reference signal and two error channels to drive four secondary sources. Using an ideal reference sensor, reduction in sound pressure level is obtained at the quiet-zone microphones averaged over the frequency range 50 to 1000 Hz with an occupant seated in the room. Two main results are presented: first for an adaptive cancelling algorithm that uses static system models, and second for the same algorithm joined with a noninvasive real-time system identification algorithm. In the first case better than 23 dB of performance is obtained if the occupant remains still through calibration and testing. In the second case, approximately 18 dB is obtained at the error microphones regardless of the motion of the occupant. PMID- 10790031 TI - Noninvasive system identification for multichannel broadband active noise control AB - Many real-world applications of active noise control are characterized by transfer functions that vary significantly and unpredictably. The controller's transfer-function models must adapt to these variations. Presented here is a class of adaptive filters that accomplish quasiperiodic system identification updates for feedforward control by using blocks of input-output histories. The algorithms form a one-dimensional family linking normalized least-mean squares (LMS) adaptive filters and block recursive least-squares, termed "block projection" algorithms, and generalize the noninvasive system identification studied by Sommerfeldt and Tichy. The system identification proceeds noninvasively, producing nonparametric impulse responses. Simulations show that the algorithm's convergence is faster than that of normalized LMS, even after the additional overhead of computing the update is taken into account. Both the multichannel generalization and application of these algorithms to system identification are novel. Simulations of the algorithms' performance using measured data are presented here, while experimental results of an implemented algorithm are contained in the companion paper. PMID- 10790032 TI - Modal control of beam flexural vibration AB - An active control system was developed to control the flexural vibrations of a beam with a modal filtering with only one secondary actuator. Segmented piezoelectric actuators and sensors were used for driving and sensing the bending beam vibrations. The primary actuator was fed by a broadband random disturbance signal in order to excite the first five modes of the structure. However, only the second to fifth modes were controlled. The control algorithm was implemented on a DSP board and the input and output signals were filtered using high order low pass filters. These filters, implemented on the DSP board avoid the degrading effect on the control performances of the higher order modes and which are not controlled. The modal filtering was achieved by computing. To this end, it is based on a previous identification procedure. This latter models, in one step, the dynamics of the structure and also the transfer function of the electronic circuits of the controller. The identified filtered modes were then used to compute the gain matrix using a LQR technique (linear quadratic regulator). Simulations of the active control were carried out and practical implementation of the control algorithms was performed. Experimental and simulation results were then compared and discussed. PMID- 10790033 TI - Broad spectrum diffusion model for room acoustics ray-tracing algorithms AB - Geometrical acoustics methods have already been transformed to account for diffusely reflecting boundaries. In randomized ray-tracing algorithms, the sound rays are either specularly reflected or scattered, according to the value of a random number which is compared with the diffusion factor. However, this method becomes inefficient if this factor depends on frequency, since the process must then be repeated for each frequency band. A method is proposed in this paper to compute all frequency components simultaneously in a single pass. The diffuse reflection model is based on the definition of a new concept: the "splitting coefficient," which can differ from the diffusion factor. First, the randomized ray-tracing method is described and the diffusion model is applied to a single diffusing surface in free field. It is shown that the results computed by the program are in accordance with theoretical results. Then, the method is shown to work properly when it is applied to more realistic enclosures: This is proved in theory and illustrated by some examples. A particular problem is the increase of the statistical error which has been solved by an appropriate control of the splitting coefficient. PMID- 10790035 TI - Beam profile measurements and simulations for ultrasonic transducers operating in air AB - This paper outlines a method that has been implemented to predict and measure the acoustic radiation generated by ultrasonic transducers operating into air in continuous wave mode. Commencing with both arbitrary surface displacement data and radiating aperture, the transmitted pressure beam profile is obtained and includes simulation of propagation channel attenuation and where necessary, the directional response of any ultrasonic receiver. The surface displacement data may be derived directly, from laser measurement of the vibrating surface, or indirectly, from finite element modeling of the transducer configuration. To validate the approach and to provide experimental measurement of transducer beam profiles, a vibration-free, draft-proof scanning system that has been installed within an environmentally controlled laboratory is described. A comparison of experimental and simulated results for piezoelectric composite, piezoelectric polymer, and electrostatic transducers is then presented to demonstrate some quite different airborne ultrasonic beam-profile characteristics. Good agreement between theory and experiment is obtained. The results are compared with those expected from a classical aperture diffraction approach and the reasons for any significant differences are explained. PMID- 10790034 TI - Acoustic load on the ear caused by headphones. AB - The standardized method for measurement of complex impedances according to ISO 10534 Part 2 is applied to the acoustic impedance of the ear with an "open-pinna" condition and with different types of headphones. The method is based on measurement of the transfer function of two microphone locations in an impedance tube and subsequent signal processing of the complex signal spectra. The termination of the tube is interpreted as ear canal entrance, while the measurement direction is, apparently, from "inside" the head towards outside. A tube which was specifically designed for this purpose works well, even though extremely small impedances must be measured. The impedances of the free pinna are similar to the "soft" end condition in the open tube, approximately following the radiation impedance of a piston into free space. The headphone impedances can be separated according to the type of headphone. In addition, the absolute impedances as the differences to the open ear compared with a number of headphones are interesting and may be starting point for further investigations. One possibility is, of course, quality control of headphones. The results are also expected to be useful for psychoacoustic research, for better understanding of sound perception, and for use in development of audio equipment. PMID- 10790036 TI - On array design for matched-field processing AB - Conventional plane-wave beamforming array design guidelines are motivated by the desire to obtain particular beampattern characteristics, such as main lobe width and side lobe levels. These design guidelines are appropriate for arrays employed for beamforming, where a plane-wave signal model is utilized to derive both the array design parameters and the beamforming algorithm. However, matched-field processing utilizes full-field acoustic propagation models to exploit the complexities of ocean acoustic propagation. As a result, there may be more appropriate design guidelines for arrays employed for matched-field processing. In this paper, general guidelines for matched-field processing array design utilizing a normal mode propagation model are proposed. Various line array configurations are evaluated with respect to source localization performance, and the results suggest that arrays designed for matched-field processing should provide a unique representation of each propagating mode along the extent of the array. Further, the empirical analyses support the guidelines suggested by the theoretical analyses and show that arrays which are far from meeting conventional beamforming array design requirements may be more than sufficient for matched field processing. PMID- 10790037 TI - Modeling the temporal behavior of distortion product otoacoustic emissions. AB - The temporal behavior of the 2f1-f2 distortion product otoacoustic emission is theoretically investigated for the case in which the lower frequency (f1) primary tone is on continuously, and the higher frequency (f2) one is pulsed on and off [e.g., Talmadge et al., J. Acoust. Soc. Am. 105, 275-292 (1999)]. On physical grounds, this behavior is expected to be characterized by various group delays associated with the propagation of (1) the f2 cochlear primary wave between the cochlear base and the primary distortion product generation region around x2 (the f2 tonotopic place), and (2) the 2f1-f2 cochlear distortion product (DP) waves between the cochlear base, the primary generation region of the distortion product, and the region around the 2f1-f2 tonotopic place where the generated apical moving DP wave is reflected toward the cochlear base [e.g., Talmadge et al., J. Acoust. Soc. Am. 104, 1517-1543 (1998)]. An approximate analytic expression is obtained for this behavior from the analysis of the Fourier integral representation of the auditory peripheral response to the primary stimuli. This expression also approximately describes the transient build-up of the components of different latencies in terms of the damping properties of the cochlear partition. It is shown that considerable caution must be applied in attempting to relate phase derivatives of the distortion product otoacoustic emissions for steady state stimuli and the physical time delays which are associated with the temporal behavior of a distortion product emission in the case of a pulsed primary. PMID- 10790038 TI - Distortion product otoacoustic emission test performance when both 2f1-f2 and 2f2 f1 are used to predict auditory status. AB - The objective of this study was to determine whether distortion product otoacoustic emission (DPOAE) test performance, defined as its ability to distinguish normal-hearing ears from those with hearing loss, can be improved by examining response and noise amplitudes at 2 f1-f2 and 2f2-f1 simultaneously. In addition, there was interest in knowing whether measurements at both DPs and for several primary frequency pairs can be used in a multivariate analysis to further optimize test performance. DPOAE and noise amplitudes were measured at 2f1-f2 and 2 f2-f1 for 12 primary levels (L2 from 10 to 65 dB SPL in 5-dB steps) and 9 pairs of primary frequencies (0.5 to 8 kHz in 1/2-octave steps). All data were collected in a sound-treated room from 70 subjects with normal hearing and 80 subjects with hearing loss. Subjects had normal middle-ear function at the time of the DPOAE test, based on standard tympanometric measurements. Measurement based stopping rules were used such that the test terminated when the noise floor around the 2 f1-f2 DP was < or = -30 dB SPL or after 32 s of artifact-free averaging, whichever occurred first. Data were analyzed using clinical decision theory in which relative operating characteristics (ROC) curves were constructed and areas under the ROC curves were estimated. In addition, test performance was assessed by selecting the criterion value that resulted in a sensitivity of 90% and determining the specificity at that criterion value. Data were analyzed using traditional univariate comparisons, in which predictions about auditory status were based only on data obtained when f2 = audiometric frequency. In addition, multivariate analysis techniques were used to determine whether test performance can be optimized by using many variables to predict auditory status. As expected, DPOAEs were larger for 2f1-f2 compared to 2 f2-f1 in subjects with normal hearing. However, noise amplitudes were smaller for 2f2-f1, but this effect was restricted to the lowest f2 frequencies. A comparison of signal-to-noise ratios (SNR) within normal-hearing ears showed that the 2f1-f2 DP was more frequently characterized by larger SNRs compared to 2f2-f1. However, there were several subjects in whom 2f2-f1 produced a larger SNR. ROC curve areas and specificities for a fixed sensitivity increased only slightly when data from both DPs were used to predict auditory status. Multivariate analyses, in which the inputs included both DPs for several primary frequency pairs surrounding each audiometric frequency, produced the highest areas and specificities. Thus, DPOAE test performance was improved slightly by examining data at two DP frequencies simultaneously. This improvement was achieved at no additional cost in terms of test time. When measurements at both DPs were combined with data obtained for several primary frequency pairs and then analyzed in a multivariate context, the best test performance was achieved. Excellent test performance (ROC) curve areas >0.95% and specificities >92% at all frequencies, including 500 Hz, were achieved for these conditions. Although the results described should be validated on an independent set of data, they suggest that the accuracy with which DPOAE measurements identify auditory status can be improved with multivariate analyses and measurements at multiple DPs. PMID- 10790039 TI - Excitotoxic effect of kainic acid on chicken otoacoustic emissions and cochlear potentials. AB - Kainic acid (KA) is a potent glutamate analog that can temporarily or permanently damage glutamatergic neurons. The purpose of the present study was to determine the short- and long-term effects of KA on chicken otoacoustic emissions and cochlear potentials. A chronic electrode was used to record the compound action potential (CAP), cochlear microphonic (CM), and the slow, positive neural potential (SPNP), a predominantly dc response. The CM, CAP, SPNP, and distortion product otoacoustic emissions (DPOAEs) were recorded before and after infusing 10 microl of a low dose (KA-L, 0.3 mM) or high dose (KA-H, 5 mM) of KA into scala tympani. KA caused a rapid and large reduction in CAP and SPNP amplitude in both the KA-H and KA-L groups; however, the CM and DPOAEs were largely unchanged. The amplitude of the CAP and SPNP in the KA-L group began to recover around 1 week post-KA, but was approximately 50% below normal at 4 weeks post-KA. In contrast, the CAP and SPNP showed no signs of recovery in the KA-H group. The results suggest that KA has no effect on the CM and DPOAEs generated by the hair cells, but selectively damages the CAP generated by the cochlear ganglion neurons. The reduction in the avian SPNP suggests that the response originates in the cochlear afferent neurons, unlike the summating potential (SP) in mammals that is generated in hair cells. PMID- 10790040 TI - Rate-intensity functions in the emu auditory nerve. AB - Rate-versus-intensity functions recorded from mammalian auditory-nerve fibers have been shown to form a continuum of shapes, ranging from saturating to straight and correlating well with spontaneous rate and sensitivity. These variations are believed to be a consequence of the interaction between the sensitivity of the hair-cell afferent synapse and the nonlinear, compressive growth of the cochlear amplifier that enhances mechanical vibrations on the basilar membrane. Little is known, however, about the cochlear amplifier in other vertebrate species. Rate-intensity functions were recorded from auditory-nerve fibers in chicks of the emu, a member of the Ratites, a primitive group of flightless birds that have poorly differentiated short and tall hair cells. Recorded data were found to be well fitted by analytical functions which have previously been shown to represent well the shapes of rate-intensity functions in guinea pigs. At the fibers' most sensitive frequencies, rate-intensity functions were almost exclusively of the sloping (80.9%) or straight (18.6%) type. Flat saturating functions, the most common type in the mammal, represented only about 0.5% of the total in the emu. Below the best frequency of each fiber, the rate intensity functions tended more towards the flat-saturating type, as is the case in mammals; a similar but weaker trend was seen above best frequency in most fibers, with only a small proportion (18%) showing the reverse trend. The emu rate-intensity functions were accepted as supporting previous evidence for the existence of a cochlear amplifier in birds, the conclusion was drawn further that the nonlinearity observed is probably due to saturation of the hair-cell transduction mechanism. PMID- 10790041 TI - Cortical, auditory, evoked potentials in response to changes of spectrum and amplitude. AB - The acoustic change complex (ACC) is a scalp-recorded negative-positive voltage swing elicited by a change during an otherwise steady-state sound. The ACC was obtained from eight adults in response to changes of amplitude and/or spectral envelope at the temporal center of a three-formant synthetic vowel lasting 800 ms. In the absence of spectral change, the group mean waveforms showed a clear ACC to amplitude increments of 2 dB or more and decrements of 3 dB or more. In the presence of a change of second formant frequency (from perceived /u/ to perceived /i/), amplitude increments increased the magnitude of the ACC but amplitude decrements had little or no effect. The fact that the just detectable amplitude change is close to the psychoacoustic limits of the auditory system augurs well for the clinical application of the ACC. The failure to find a condition under which the spectrally elicited ACC is diminished by a small change of amplitude supports the conclusion that the observed ACC to a change of spectral envelope reflects some aspect of cortical frequency coding. Taken together, these findings support the potential value of the ACC as an objective index of auditory discrimination capacity. PMID- 10790042 TI - Conditioning-induced protection from impulse noise in female and male chinchillas. AB - Sound conditioning (pre-exposure to a moderate-level acoustic stimulus) can induce resistance to hearing loss from a subsequent traumatic exposure. Most sound conditioning experiments have utilized long-duration tones and noise at levels below 110 dB SPL as traumatic stimuli. It is important to know if sound conditioning can also provide protection from brief, high-level stimuli such as impulses produced by gunfire, and whether there are differences between females and males in the response of the ear to noise. In the present study, chinchillas were exposed to 95 dB SPL octave band noise centered at 0.5 kHz for 6 h/day for 5 days. After 5 days of recovery, they were exposed to simulated M16 rifle fire at a level of 150 dB peak SPL. Animals that were sound conditioned showed less hearing loss and smaller hair cell lesions than controls. Females showed significantly less hearing loss than males at low frequencies, but more hearing loss at 16 kHz. Cochleograms showed slightly less hair cell loss in females than in males. The results show that significant protection from impulse noise can be achieved with a 5-day conditioning regimen, and that there are consistent differences between female and male chinchillas in the response of the cochlea to impulse noise. PMID- 10790044 TI - Contributions of suppression and excitation to simultaneous masking: effects of signal frequency and masker-signal frequency relation. AB - This study investigated the contributions of suppression and excitation to simultaneous masking for a range of masker frequencies both below and above three different signal frequencies (750, 2000, and 4850 Hz). A two-stage experiment was employed. In stage I, the level of each off-frequency simultaneous masker necessary to mask a signal at 10 or 30 dB sensation level was determined. In stage II, three different forward-masking conditions were tested: (1) an on frequency condition, in which the signals in stage I were used to mask probes of the same frequency; (2) an off-frequency condition, in which the off-frequency maskers (at the levels determined in stage I) were used to mask the probes; and (3) a combined condition, in which the on- and off-frequency maskers were combined to mask the probes. If the off-frequency maskers simultaneously masked the signal via spread of excitation in stage I, then the off-frequency and combined maskers should produce considerable forward masking in stage II. If, on the other hand, they masked via suppression, they should produce little or no forward masking. The contribution of suppression was found to increase with increasing signal frequency; it was absent at 750 Hz, but dominant at 4850 Hz. These results have implications for excitation pattern analyses and are consistent with stronger nonlinear processing at high rather than at low frequencies. PMID- 10790043 TI - Enhancements of the edges of temporal masking functions by complex patterns of overshoot and undershoot. AB - The purpose of this report is to present new data that provide a novel perspective on temporal masking, different from that found in the classical auditory literature on this topic. Specifically, measurement conditions are presented that minimize rather than maximize temporal spread of masking for a gated (200-ms) narrow-band (405-Hz-wide) noise masker logarithmically centered at 2500 Hz. Masked detection thresholds were measured for brief sinusoids in a two interval, forced-choice (21FC) task. Detection was measured at each of 43 temporal positions within the signal observation interval for the sinusoidal signal presented either preceding, during, or following the gating of the masker, which was centered temporally within each 500-ms observation interval. Results are presented for three listeners; first, for detection of a 1900-Hz signal across a range of masker component levels (0-70 dB SPL) and, second, for masked detection as a function of signal frequency (fs = 500-5000 Hz) for a fixed masker component level (40 dB SPL). For signals presented off-frequency from the masker, and at low-to-moderate masker levels, the resulting temporal masking functions are characterized by sharp temporal edges. The sharpness of the edges is accentuated by complex patterns of temporal overshoot and undershoot, corresponding with diminished and enhanced detection, respectively, at both masker onset and offset. This information about the onset and offset timing of the gated masker is faithfully represented in the temporal masking functions over the full decade range of signal frequencies (except for fs=2500 Hz presented at the center frequency of the masker). The precise representation of the timing information is remarkable considering that the temporal envelope characteristics of the gated masker are evident in the remote masking response at least two octaves below the frequencies of the masker at a cochlear place where little or no masker activity would be expected. This general enhancement of the temporal edges of the masking response is reminiscent of spectral edge enhancement by lateral suppression/inhibition. PMID- 10790045 TI - Dichotic pitches as illusions of binaural unmasking. III. The existence region of the Fourcin pitch. AB - Two experiments explored the existence region of the Fourcin pitch. In each experiment, detectability was assessed by measuring listeners' ability to discriminate pitch changes. In the first experiment, the detectability of the pitch was measured as a function of the number of noises used to generate it. In the second experiment, the pitch was generated using two noises with equal and opposite interaural delays and detectability was measured as a function of the difference between these two delays, and thus of the perceived pitch height. In each case, the experimental results were compared with the predictions produced by a model of binaural unmasking, based on equalization cancellation, that had been designed to recover broadband sounds, such as speech, from interfering noise [Culling and Summerfield, J. Acoust. Soc. Am. 98, 785-797 (1995)]. The model accurately predicted the results from experiment 1, but failed to show an adequate decline in performance for small differences in interaural delay (corresponding to higher perceived pitches) in experiment 2. A revised model, based on similar principles, but using data on listeners' sensitivity to interaural decorrelation, rather than an equalization-cancellation mechanism, was able to predict the results of both experiments successfully. PMID- 10790047 TI - Influence of spatial and temporal coding on auditory gap detection. AB - This study investigated the effect on gap detection of perceptual channels, hypothesized to be tuned to spatial location or fundamental frequency (f0). Thresholds were measured for the detection of a silent temporal gap between two markers. In the first experiment, the markers were broadband noise, presented either binaurally or monaurally. In the binaural conditions, the markers were either diotic, or had a 640-micros interaural time difference (ITD) or a 12-dB interaural level difference (ILD). Reversing the ITD across the two markers had no effect on gap detection relative to the diotic condition. Reversing the ILD across the two markers produced a marked deterioration in performance. However, the same deterioration was observed in the monaural conditions when a 12-dB level difference was introduced between the two markers. The results provide no evidence for the role of spatially tuned neural channels in gap detection. In the second experiment, the markers were harmonic tone complexes, filtered to contain only high, unresolved harmonics. Using complexes with a fixed spectral envelope, where the f0 (of 140 or 350 Hz) was different for the two markers, produced a deterioration in performance, relative to conditions where the f0 remained the same. A larger deterioration was observed when the two markers occupied different spectral regions but had the same f0. This supports the idea that peripheral coding is dominant in determining gap-detection thresholds when the two markers differ along any physical dimension. Higher-order neural coding mechanisms of f0 and spatial location seem to play a smaller role and no role, respectively. PMID- 10790046 TI - Auditory thresholds in a sound-producing electric fish (Pollimyrus): behavioral measurements of sensitivity to tones and click trains. AB - In this report we present the first behavioral measurements of auditory sensitivity for Pollimyrus adspersus. Pollimyrus is an electric fish (Mormyridae) that uses both electric and acoustic signals for communication. Tone detection was assessed from the fish's electric organ discharge rate. Suprathreshold tones usually evoked an accelerated rate in naive animals. This response (rate modulation > or =25%) was maintained in a classical conditioning paradigm by presenting a weak electric current near the offset of 3.5-s tone bursts. An adaptive staircase procedure was used to find detection thresholds at frequencies between 100 and 1700 Hz. The mean audiogram from six individuals revealed high sensitivity in the 200-900 Hz range, with the best thresholds near 500 Hz (66.5+/ 4.2 SE dB re: 1 microPa). Sensitivity declined slowly (about 20 dB/octave) above and below this sensitivity maximum. Sensitivity fell off rapidly above 1 kHz (about 60 dB/octave) and no responses were observed at 5 kHz. This behavioral sensitivity matched closely the spectral content of the sounds that this species produced during courtship. Experiments with click trains showed that sensitivity (about 83-dB peak) was independent of inter-click-interval, within the 10-100 ms range. PMID- 10790048 TI - Multichannel speech intelligibility and talker recognition using monaural, binaural, and three-dimensional auditory presentation. AB - In a 3D auditory display, sounds are presented over headphones in a way that they seem to originate from virtual sources in a space around the listener. This paper describes a study on the possible merits of such a display for bandlimited speech with respect to intelligibility and talker recognition against a background of competing voices. Different conditions were investigated: speech material (words/sentences), presentation mode (monaural/binaural/3D), number of competing talkers (1-4), and virtual position of the talkers (in 45 degrees-steps around the front horizontal plane). Average results for 12 listeners show an increase of speech intelligibility for 3D presentation for two or more competing talkers compared to conventional binaural presentation. The ability to recognize a talker is slightly better and the time required for recognition is significantly shorter for 3D presentation in the presence of two or three competing talkers. Although absolute localization of a talker is rather poor, spatial separation appears to have a significant effect on communication. For either speech intelligibility, talker recognition, or localization, no difference is found between the use of an individualized 3D auditory display and a general display. PMID- 10790049 TI - Lip-jaw and tongue-jaw coordination during rate-controlled syllable repetitions. AB - The present study investigated the relationship between functionally relevant compound gestures and single-articulator component movements of the jaw and the constrictors lower lip and tongue tip during rate-controlled syllable repetitions. In nine healthy speakers, the effects of speaking rate (3 vs 5 Hz), place of articulation, and vowel type during stop consonant-vowel repetitions (/pa/, /pi/, /ta/, /ti/) on the amplitude and peak velocity of differential jaw and constrictor opening-closing movements were measured by means of electromagnetic articulography. Rather than homogeneously scaled compound gestures, the results suggest distinct control mechanisms for the jaw and the constrictors. In particular, jaw amplitude was closely linked to vowel height during bilabial articulation, whereas the lower lip component amplitude turned out to be predominantly rate sensitive. However, the observed variability across subjects and conditions does not support the assumption that single-articulator gestures directly correspond to basic phonological units. The nonhomogeneous effects of speech rate on articulatory subsystem parameters indicate that single structures are differentially rate sensitive. On average, an increase in speech rate resulted in a more or less proportional increase of the steepness of peak velocity/amplitude scaling for jaw movements, whereas the constrictors were less rate sensitive in this respect. Negative covariation across repetitions between jaw and constrictor amplitudes has been considered an indicator of motor equivalence. Although significant in some cases, such a relationship was not consistently observed across subjects. Considering systematic sources of variability such as vowel height, speech rate, and subjects, jaw-constrictor amplitude correlations showed a nonhomogeneous pattern strongly depending on place of articulation. PMID- 10790050 TI - Piano hammers and their force compression characteristics: does a power law make sense? AB - We have studied the force characteristics of a collection of piano hammers, through studies of the acceleration of the hammer head, the force due to the compression of the hammer felt, Fh, and the associated bending of the hammer shank which occurs when a hammer strikes a rigid object. By integration of the acceleration one can estimate the compression of the felt, and thus obtain a force-compression Fh(z) relationship; our results for this function are compared with previous experiments and theoretical models. Close examination of our findings suggests that bending of the hammer shank, and also the time required for the force to be transmitted from the outer edge of the hammer to its core, play significant roles in the hammer dynamics. The data are used to estimate the quantitative impact of these effects on the derived Fh(z) relation. The implications for physical modeling of a piano tone are briefly discussed. PMID- 10790051 TI - Why pinnipeds don't echolocate. AB - Odontocete cetaceans have evolved a highly advanced system of active biosonar. It has been hypothesized that other groups of marine animals, such as the pinnipeds, possess analogous sound production, reception, and processing mechanisms that allow for underwater orientation using active echolocation. Despite sporadic investigation over the past 30 years, the accumulated evidence in favor of the pinniped echolocation hypothesis is unconvincing. We argue that an advanced echolocation system is unlikely to have evolved in pinnipeds primarily because of constraints imposed by the obligate amphibious functioning of the pinniped auditory system. As a result of these constraints, pinnipeds have not developed highly acute, aquatic, high frequency sound production or reception systems required for underwater echolocation. Instead, it appears that pinnipeds have evolved enhanced visual, tactile, and passive listening skills. The evolutionary refinement of alternative sensory systems allows pinnipeds to effectively forage, navigate, and avoid predators under water despite the lack of active biosonar capabilities. PMID- 10790053 TI - Oscillations of polymeric microbubbles: effect of the encapsulating shell AB - A model for the oscillation of gas bubbles encapsulated in a thin shell has been developed. The model depends on viscous and elastic properties of the shell, described by thickness, shear modulus, and shear viscosity. This theory was used to describe an experimental ultrasound contrast agent from Nycomed, composed of air bubbles encapsulated in a polymer shell. Theoretical calculations were compared with measurements of acoustic attenuation at amplitudes where bubble oscillations are linear. A good fit between measured and calculated results was obtained. The results were used to estimate the viscoelastic properties of the shell material. The shell shear modulus was estimated to between 10.6 and 12.9 MPa, the shell viscosity was estimated to between 0.39 and 0.49 Pas. The shell thickness was 5% of the particle radius. These results imply that the particles are around 20 times more rigid than free air bubbles, and that the oscillations are heavily damped, corresponding to Q-values around 1. We conclude that the shell strongly alters the acoustic behavior of the bubbles: The stiffness and viscosity of the particles are mainly determined by the encapsulating shell, not by the air inside. PMID- 10790052 TI - Vocal control of acoustic information for sonar discriminations by the echolocating bat, Eptesicus fuscus. AB - This study aimed to determine whether bats using frequency modulated (FM) echolocation signals adapt the features of their vocalizations to the perceptual demands of a particular sonar task. Quantitative measures were obtained from the vocal signals produced by echolocating bats (Eptesicus fuscus) that were trained to perform in two distinct perceptual tasks, echo delay and Doppler-shift discriminations. In both perceptual tasks, the bats learned to discriminate electronically manipulated playback signals of their own echolocation sounds, which simulated echoes from sonar targets. Both tasks utilized a single-channel electronic target simulator and tested the bat's in a two-alternative forced choice procedure. The results of this study demonstrate changes in the features of the FM bats' sonar sounds with echolocation task demands, lending support to the notion that this animal actively controls the echo information that guides its behavior. PMID- 10790054 TI - Experimental investigation of the pulse inversion technique for imaging ultrasound contrast agents. AB - The application of ultrasound contrast agents aims to detect low velocity blood flow in the microcirculation. To enhance discrimination between tissue and blood containing the contrast agent, harmonic imaging is used. Harmonic imaging requires the application of narrow-band signals and is obscured by high levels of native harmonics generated in an intervening medium. To improve discrimination between contrast agent and native harmonics, a pulse inversion technique has been proposed. Pulse inversion allows wide-band signals, thus preserving the axial resolution. The present study examines the interference of native harmonics and discusses the practical difficulties of wide-band pulse inversion measurements of harmonics by a single transducer. Native harmonics are not eliminated by pulse inversion. Furthermore, only even harmonics remain and are amplified by 6 dB, alleviating the requirement for selective filtering. Finally, it is shown that the contaminating third harmonic contained in the square wave activation signal leaks through in the emitted signal. The spectral location of the contaminating third harmonic is governed by the transducer spectral characteristics while the location of the native and contrast agent second harmonics is not. Thus the contaminating third harmonic and the native and contrast agent second harmonics may overlap and interfere. Optimal discrimination requires a balance between maximal sensitivity for the second harmonic at reception and minimal interference from the contaminating third harmonic. PMID- 10790055 TI - Profile analysis of harmonic complexes: effects of mistuning the target. PMID- 10790057 TI - Intervening O vi Quasar Absorption Systems at Low Redshift: A Significant Baryon Reservoir. AB - Far-UV echelle spectroscopy of the radio-quiet QSO H1821+643 (zem=0.297), obtained with the Space Telescope Imaging Spectrograph (STIS) at approximately 7 km s-1 resolution, reveals four definite O vi absorption-line systems and one probable O vi absorber at 0.15-0.3) of the stellar content of omega Cen and show anomalous abundances of s-process elements (as Ba and Zr) as well; (2) they are physical members of the omega Cen system; (3) they comprise approximately 5% of the stars of the whole system; and (4) this component and the metal intermediate one (-0.4>&sqbl0;Ca&solm0;H&sqbr0;>-1) have been found to share the same spatial distribution, both of them differing significantly from the most metal-poor one (&sqbl0;Ca&solm0;H&sqbr0;<-1). This last piece evidence supports the hypothesis that metal-rich components could belong to an independent (proto?) stellar system captured in the past by omega Cen. PMID- 10790078 TI - Formation of the Galactic Stellar Halo: Origin of the Metallicity-Eccentricity Relation. AB - Motivated by the recently improved knowledge on the kinematic and chemical properties of the Galactic metal-poor stars, we present the numerical simulation for the formation of the Galactic stellar halo to interpret the observational results. As a model for the Galaxy contraction, we adopt the currently standard theory of galaxy formation based on the hierarchical assembly of the cold dark matter fluctuations. We find, for the simulated stars with &sqbl0;Fe&solm0;H&sqbr0;/=0.5 AU), no later than a few times 107 yr of birth. PMID- 10790081 TI - VLA Imaging of the Disk Surrounding the Nearby Young Star TW Hydrae. AB - The TW Hydrae system is perhaps the closest analog to the early solar nebula. We have used the Very Large Array to image TW Hya at wavelengths of 7 mm and 3.6 cm with resolutions of 0&farcs;1 ( approximately 5 AU) and 1&farcs;0 ( approximately 50 AU), respectively. The 7 mm emission is extended and appears dominated by a dusty disk of radius greater than 50 AU surrounding the star. The 3.6 cm emission is unresolved and likely arises from an ionized wind or gyrosynchrotron activity. The dust spectrum and spatially resolved 7 mm images of the TW Hya disk are fitted by a simple model with temperature and surface density described by radial power laws, T&parl0;r&parr0;~r-0.5 and Sigma&parl0;r&parr0;~r-1. These properties are consistent with an irradiated gaseous accretion disk of mass approximately 0.03 M middle dot in circle with an accretion rate approximately 10-8 M middle dot in circle yr-1 and viscosity parameter alpha=0.01. The estimates of mass and mass accretion rates are uncertain since the gas-to-dust ratio in the TW Hya disk may have evolved from the standard interstellar value. PMID- 10790082 TI - Correcting Radial Velocities for Long-Term Magnetic Activity Variations. AB - We study stars in the Lick planetary survey for correlations between simultaneous measurements of high-precision radial velocities vr and magnetic activity (as measured in an SIR emission index from Ca ii lambda8662). We find significant correlations in approximately 30% of the stars. After removing linear trends between SIR and vr, we find that the dispersion in vr in these stars is decreased by an average of 17%, or approximately 45% of the dispersion above the measurement noise. F stars and less active stars with variable Ca ii H and K lines are the most successfully corrected. The magnitude of the slope of the SIR versus vr relations increases proportional to vsini and (excepting M dwarfs) tends to decrease with decreasing Teff. We argue that the main cause of these effects is modification of the mean line bisector shape brought on by long-term, magnetic activity-induced changes in the surface brightness and convective patterns. The correlations can be used to partially correct vr data for the effects of long-term activity variations, potentially permitting study of planets around some (higher mass) younger stars and planets producing smaller stellar reflex velocities. PMID- 10790083 TI - Low-Energy Anomalous Cosmic Rays in the Ecliptic Plane: 1-5 AU. AB - Anomalous cosmic-ray (ACR) oxygen (O) fluxes in the energy range 0.5-5.0 MeV nucleon-1 were measured in the ecliptic plane at 1 and 5 AU in 1997 and 1998. Using measurements from essentially identical instruments on the Advanced Composition Explorer (ACE) and Ulysses spacecraft, we were able to monitor the decrease with time of the ACR O fluxes in the inner heliosphere during the onset of solar cycle 23. The data obtained from three time intervals that are relatively undisturbed by solar activity during this period show that the e folding time is approximately 140+/-40 days for the disappearance of 2-5 MeV nucleon-1 ACR oxygen from the ecliptic plane at 5 AU. PMID- 10790084 TI - Semiconvection and Overshooting: Schwarzschild and Ledoux Criteria Revisited. AB - We show that the Schwarzschild formalism, if accepted as the description of the structure of a semiconvective region, necessarily implies overshooting. The Ledoux formalism (if accepted) can be used without overshooting, provided a new relation for the mixing-length parameter alpha is satisfied. Should the latter not be satisfied, the Ledoux formalism would also require overshooting, that is, a nonlocal convection model. PMID- 10790085 TI - Detection of Legionella species in respiratory specimens using PCR with sequencing confirmation. AB - Legionella spp. are a common cause of community-acquired respiratory tract infections and an occasional cause of nosocomial pneumonia. A PCR method for the detection of legionellae in respiratory samples was evaluated and was compared to culture. The procedure can be performed in 6 to 8 h with a commercially available DNA extraction kit (Qiagen, Valencia, Calif.) and by PCR with gel detection. PCR is performed with primers previously determined to amplify a 386-bp product within the 16S rRNA gene of Legionella pneumophila. We can specifically detect the clinically significant Legionella species including L. pneumophila, L. micdadei, L. longbeachae, L. bozemanii, L. feeleii, and L. dumoffii. The assay detects 10 fg (approximately two organisms) of legionella DNA in each PCR. Of 212 clinical specimens examined by culture, 100% of the culture-positive samples (31 of 31) were positive by this assay. By gel detection of amplification products, 12 of 181 culture-negative samples were positive for Legionella species by PCR, resulting in 93% specificity. Four of the 12 samples with discrepant results (culture negative, PCR positive) were confirmed to be positive for Legionella species by sequencing of the amplicons. The legionella-specific PCR assay that is described demonstrates high sensitivity and high specificity for routine detection of legionellae in respiratory samples. PMID- 10790086 TI - Evaluation of the PASCO strep plus broth microdilution antimicrobial susceptibility panels for testing Streptococcus pneumoniae and other Streptococcal species. AB - Antimicrobial resistance continues to increase worldwide among isolates of Streptococcus pneumoniae and other species of streptococci. Increasing rates of penicillin resistance, particularly in viridans group streptococci, and resistance to multiple classes of antimicrobial agents, including beta-lactams, macrolides, and fluoroquinolones, in pneumococci have increased the importance of having accurate antimicrobial susceptibility testing results for guiding therapy. One commercial method of assessing resistance in streptococci is the PASCO Strep Plus panel. This broth microdilution-based method has recently been expanded to include a variety of newer antimicrobial agents. Therefore, we compared the results of the new PASCO Strep Plus panels for 26 antimicrobial agents against the results generated using the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution reference method for 75 pneumococci and 68 other streptococcal isolates. Only 4 (0.2%) very major errors (all with pneumococci and each with a different antimicrobial agent) were observed. There were 5 (0.3%) major errors observed with pneumococci (each with a different antimicrobial agent), but only 1 major error with nonpneumococcal streptococci. All of the very major and major errors resolved on retesting. Of the 65 (3.9%) and 17 (1.6%) minor errors observed with pneumococci and other streptococci, respectively, all were within 1 dilution of the broth microdilution reference MIC result. Thus, the PASCO Strep Plus panel has comparable accuracy to the NCCLS broth microdilution reference method. PMID- 10790087 TI - Rapid identification and differentiation of Bartonella species using a single step PCR assay. AB - Five species of Bartonella have been reported to infect humans and cause a variety of diseases that can be difficult to diagnose. Four species of Bartonella have been reported to infect cats and dogs, and two of these species are considered to be zoonotic pathogens. Diagnosis of Bartonella infections is hampered by the slow, fastidious growth characteristics of Bartonella species. We report on the development of a single-step PCR-based assay for the detection and differentiation of medically relevant Bartonella species. PCR-mediated amplification of the 16S-23S rRNA intergenic region resulted in a product of a unique size for each Bartonella species, thereby allowing differentiation without the necessity of restriction fragment length polymorphism analysis or sequencing of the amplified product. The ability of the single-step PCR assay to differentiate between Bartonella species was determined with characterized isolates and blood samples from animals known to be infected with either Bartonella henselae, B. clarridgeiae, or B. vinsonii subsp. berkhoffii. The sensitivity of the single-step PCR assay relative to that of in vitro culture was determined with blood samples from B. henselae-infected cats. B. henselae target DNA was amplified from 100% of samples with greater than 50 CFU/ml and 80% of samples with 10 to 30 CFU/ml. The single-step assay described in the report expedites PCR-based detection and differentiation of medically relevant Bartonella species. PMID- 10790088 TI - Identification of a novel DNA probe for strain typing Mycobacterium bovis by restriction fragment length polymorphism analysis. AB - Bovine tuberculosis caused by Mycobacterium bovis remains a significant disease of farmed cattle in many countries despite ongoing tuberculosis eradication programs. Molecular typing methods such as restriction fragment length polymorphism (RFLP) analysis and spoligotyping have been used to identify related herd breakdowns in an attempt to identify more precisely the route of infection into cattle herds and to trace the transmission of bovine tuberculosis. A recent geographical survey of Irish M. bovis isolates demonstrated that a significant proportion of isolates ( approximately 20%) exhibit a common strain type, limiting the value of current strain typing methods as an epidemiological tool. We have identified and cloned a region of the M. bovis genome, pUCD, which generates a clear, highly polymorphic banding pattern when used as an RFLP probe on AluI restriction-digested M. bovis genomic DNA and which effectively subdivides this common strain type. When used to type 60 Irish M. bovis isolates, pUCD exhibited greater discriminatory power than the commonly used mycobacterial RFLP probes IS6110, PGRS, and DR and detected an equivalent number of strain types to a combination of these three probes. pUCD also detected significantly more strain types than the spoligotyping technique, while maintaining a high level of concordance between epidemiologically related and unrelated herd breakdowns. The polymorphic element within pUCD remains to be fully characterized, however the potential for this probe to greatly decrease the workload necessary to genotype M. bovis by RFLP analysis is compelling. PMID- 10790089 TI - Use of clindamycin disks To detect macrolide resistance mediated by ermB and mefE in Streptococcus pneumoniae isolates from adults and children. AB - We studied 198 macrolide-resistant S. pneumoniae isolates obtained from adults and children to evaluate whether 2-microgram clindamycin disks can distinguish between isolates manifesting ermB- versus mefE-mediated resistance to clarithromycin and to determine the relative frequency with which each resistance mechanism occurred in these populations. The mefE gene was predominant among 109 isolates from children, occurring in 73.4% versus 50.6% of 89 isolates from adults. Three isolates (1.5%) did not amplify either gene. Among 125 mefE(+) isolates, the MIC of clarithromycin at which 90% of the isolates tested were inhibited, determined by Etest, was 32 microgram/ml versus >256 microgram/ml in 70 ermB(+) isolates. All ermB(+) isolates were highly resistant to clindamycin (MICs >256 microgram/ml), whereas all mefE(+) isolates were susceptible to clindamycin using the 2-microgram disk. Testing S. pneumoniae from the respiratory tract for susceptibility to clindamycin by agar disk diffusion is an easy and inexpensive method to estimate the frequency of resistance mediated by ermB in specific patient populations. Macrolide resistance mediated by ermB is usually of greater magnitude than that due to mefE. Clinical studies are needed to determine the significance of high- versus low-level macrolide resistance in S. pneumoniae. PMID- 10790090 TI - Serologic diagnosis of Lyme borreliosis by using enzyme-linked immunosorbent assays with recombinant antigens. AB - Class-specific enzyme-linked immunosorbent assays (ELISAs) with purified recombinant antigens of Borrelia burgdorferi sensu stricto and Western blot analyses with whole cells of this spirochete were used to test human sera to determine which antigens were diagnostically important. In analyses for immunoglobulin M (IgM) antibodies, 14 (82%) of 17 serum samples from persons who had erythema migrans reacted positively by an ELISA with one or more recombinant antigens. There was frequent antibody reactivity to protein 41-G (p41-G), outer surface protein C (OspC), and OspF antigens. In an ELISA for IgG antibodies, 13 (87%) of 15 serum samples had antibodies to recombinant antigens; reactivity to p22, p39, p41-G, OspC, and OspF antigens was frequent. By both ELISAs, serum specimens positive for OspB, OspE, and p37 were uncommon. Analyses of sera obtained from persons who were suspected of having human granulocytic ehrlichiosis (HGE) but who lacked antibodies to ehrlichiae revealed IgM antibodies to all recombinant antigens of B. burgdorferi except OspB and IgG antibodies to all antigens except OspE. Immunoblotting of sera from the study group of individuals suspected of having HGE reaffirmed antibody reactivity to multiple antigens of B. burgdorferi. There was minor cross-reactivity when sera from healthy subjects or persons who had syphilis, oral infections, or rheumatoid arthritis were tested by ELISAs with p37, p41-G, OspB, OspC, OspE, and OspF antigens. Although the results of class-specific ELISAs with recombinant antigens were comparable to those recorded for assays with whole-cell antigen and for individuals with confirmed clinical diagnoses of Lyme borreliosis, immunoblotting is still advised as an adjunct procedure, particularly when there are low antibody titers by an ELISA. PMID- 10790091 TI - Molecular epidemiology of Staphylococcus epidermidis in a neonatal intensive care unit over a three-year period. AB - Coagulase-negative staphylococci, especially Staphylococcus epidermidis, are increasingly important nosocomial pathogens, particularly in critically ill neonates. A 3-year prospective surveillance of nosocomial infections in a neonatal intensive care unit (NICU) was performed by traditional epidemiologic methods as well as molecular typing of microorganisms. The aims of the study were (i) to quantify the impact of S. epidermidis on NICU-acquired infections, (ii) to establish if these infections are caused by endemic clones or by incidentally occurring bacterial strains of this ubiquitous species, (iii) to evaluate the use of different methods for the epidemiologic typing of the isolates, and (iv) to characterize the occurrence and the spread of staphylococci with decreased glycopeptide susceptibility. Results confirmed that S. epidermidis is one of the leading causes of NICU-acquired infections and that the reduced glycopeptide susceptibility, if investigated by appropriate detection methods such as population analysis, is more common than is currently realized. Typing of isolates, which can be performed effectively through molecular techniques such as pulsed-field gel electrophoresis but not through antibiograms, showed that many of these infections are due to clonal dissemination and, thus, are potentially preventable by strict adherence to recommended infection control practices and the implementation of programs aimed toward the reduction of the unnecessary use of antibiotics. These strategies are also likely to have a significant impact on the frequency of the reduced susceptibility of staphylococci to glycopeptides, since this phenomenon appears to be determined either by more resistant clones transmitted from patient to patient or, to a lesser extent, by strains that become more resistant as a result of antibiotic pressure. PMID- 10790092 TI - Contamination and sensitivity issues with a real-time universal 16S rRNA PCR. AB - A set of universal oligonucleotide primers specific for the conserved regions of the eubacterial 16S rRNA gene was designed for use with the real-time PCR Applied Biosystems 7700 (TaqMan) system. During the development of this PCR, problems were noted with the use of this gene as an amplification target. Contamination of reagents with bacterial DNA was a major problem exacerbated by the highly sensitive nature of the real-time PCR chemistry. This was compounded by the use of a small amplicon of approximately 100 bases, as is necessary with TaqMan chemistry. In an attempt to overcome this problem, several methodologies were applied. Certain treatments were more effective than others in eliminating the contaminating DNA; however, to achieve this there was a decrease in sensitivity. With UV irradiation there was a 4-log reduction in PCR sensitivity, with 8 methoxypsoralen activity facilitated by UV there was between a 5- and a 7-log reduction, and with DNase alone and in combination with restriction digestion there was a 1.66-log reduction. Restriction endonuclease treatment singly and together did not reduce the level of contaminating DNA. Without the development of ultrapure Taq DNA polymerase, ultrapure reagents, and plasticware guaranteed to be free of DNA, the implementation of a PCR for detection of eubacterial 16S rRNA with the TaqMan system will continue to be problematical. PMID- 10790093 TI - Detection of and discrimination between gram-positive and gram-negative bacteria in intraocular samples by using nested PCR. AB - A nested PCR protocol has been developed for the detection of and discrimination between 14 species of gram-positive and -negative bacteria in samples of ocular fluids. First-round PCR with pan-bacterial oligonucleotide primers, based on conserved sequences of the 16S ribosomal gene, was followed by a gram-negative organism-specific PCR, which resulted in a single 985-bp amplification product, and a multiplex PCR which resulted in two PCR products: a 1,025 bp amplicon (all bacteria) and a 355 bp amplicon (gram-positive bacteria only). All products were detected by gel electrophoresis. The sensitivity of the assay was between 10 fg and 1 pg of bacterial DNA, depending on the species tested, equivalent to between 24 and 4 live bacteria spiked in water. The identification was complete in 3.5 h. The molecular techniques were subsequently applied to four samples of intraocular fluid, (three vitreous and one aqueous) from three patients with clinical signs of bacterial endophthalmitis (test samples) and two samples of vitreous from a patient with chronic intraocular inflammation (control samples). In all culture positive samples (two of three vitreous and one of one aqueous), a complete concordance was observed between molecular methods and culture results. PCR correctly identified the gram stain classification of the organisms. The bacterial etiology was also identified in a culture-negative patient with clinical history and signs highly suggestive of bacterial endophthalmitis. Furthermore, control samples from a patient with chronic intraocular inflammation remained PCR negative. In summary, this protocol has demonstrated potential as a rapid diagnostic test in confirming the diagnosis of infection and also determining the Gram status of bacteria with high specificity and sensitivity. PMID- 10790094 TI - Zinc salts inactivate clinical isolates of herpes simplex virus in vitro. AB - Using a standard plaque assay and clinical isolates of herpes simplex virus (HSV), we have tested the ability of zinc salts to inactivate HSV. Virus was treated by incubation at 37 degrees C with zinc salts in morpholinepropanesulfonic acid-buffered culture medium and was then diluted and plated onto CV-1 cells for detection and quantitation of remaining infectious virus. Of 10 randomly chosen clinical isolates (five HSV type 1 [HSV-1] isolates and five HSV-2 isolates), seven were inactivated >98% by treatment in vitro with 50 mM zinc gluconate for 2 h and nine were inactivated >97% by treatment with zinc lactate. The effect was concentration dependent. With an HSV-1 isolate, 50 mM zinc gluconate or zinc lactate caused 100% inactivation, 15 mM caused 98 to 99% inactivation, and 5 mM caused 63 to 86% inactivation. With an HSV-2 isolate, 50 and 15 mM zinc gluconate caused 30% inactivation and 5 and 1 mM caused less than 9% inactivation, whereas 50 and 15 mM zinc lactate caused greater than 92% inactivation and 5 and 1 mM caused 37 and 26% inactivation, respectively. The ability of the zinc salts to inactivate HSV was not related to pH in the pH range of 6.1 to 7.6 since inactivation by zinc gluconate or zinc lactate in that pH range was 99.7 to 100% with a 2-h treatment with 50 mM zinc salt. Short (5-min) treatments of selected isolates with zinc gluconate, zinc lactate, zinc acetate, or zinc sulfate yielded inactivation rates of 0 to 55%. PMID- 10790095 TI - Distribution of genes encoding putative transmissibility factors among epidemic and nonepidemic strains of Burkholderia cepacia from cystic fibrosis patients in the United Kingdom. AB - In the last 15 years, Burkholderia cepacia has emerged as a significant pathogen in cystic fibrosis (CF) patients, mainly due to the severity of infection observed in a subset of patients and the fear of transmission of the organism to noncolonized patients. Although patients who deteriorate rapidly cannot be predicted by microbiological characteristics, three genetic markers have been described for strains that spread between patients. These are the cblA gene, encoding giant cable pili; a hybrid of two insertion sequences, IS1356 and IS402; and a 1.4-kb open reading frame known as the B. cepacia epidemic strain marker (BCESM). The latter two are of unknown function. An epidemic strain lineage was previously identified among CF patients in the United Kingdom that apparently had spread from North America and that was characterized by a specific random amplified polymorphic DNA (RAPD) pattern. We searched for the described genetic markers using specific PCR assays with 117 patient isolates of B. cepacia from 40 United Kingdom hospitals. Isolates were grouped according to genomovar and epidemic strain lineage RAPD pattern with a 10-base primer, P272. A total of 41 isolates from patients in 12 hospitals were classified as the epidemic strain, and 40 of these were distributed in genomovars IIIa (11 isolates), IIIb (1 isolate), and IIIc (28 isolates). All isolates of the epidemic strain were positive for the cblA gene and BCESM, but two lacked the insertion sequence hybrid. None of the 76 sporadic isolates contained cblA or the insertion sequence hybrid, but 11 of them were positive for BCESM. Nonepidemic isolates were distributed among genomovars I or IV (9), II (49), IIIa (11), IIIb (3), and IIIc (4). There were three clusters of cross-infection (one involving two patients and two involving three patients) with isolates of genomovar II. We conclude that in the United Kingdom, a single clonal lineage has spread between and within some hospitals providing care for CF patients. The presence of the cblA gene is the most specific marker for the epidemic strain. We recommend that all isolates of B. cepacia from CF patients should be screened by PCR to influence segregation and infection control strategies. PMID- 10790096 TI - Identification of enterotoxigenic Escherichia coli harboring longus type IV pilus gene by DNA amplification. AB - DNA amplification of lngA, the structural gene of longus type IV pilus produced by human enterotoxigenic Escherichia coli (ETEC) was achieved by the use of specific oligonucleotide primers designed from the nucleotide sequence of lngA. A 630-bp fragment representing the entire lngA gene was amplified in eight prototype strains previously characterized as longus positive. Five ETEC strains producing colonization factor antigen III (CFA III) (also a type IV pilus) were also positive by PCR, confirming the DNA homology between CFA III and longus. None of the non-ETEC and non-E. coli enteropathogens studied showed the 0.63-kbp amplicon. The procedure thus detected only ETEC strains harboring type IV pili genes with or without other colonization factors. Except for five lngA PCR positive, probe-positive strains, all lngA PCR-positive strains produced the pilin as demonstrated by immunoblotting. To test the amplification procedure in a clinical setting, a collection of 264 fresh clinical E. coli strains isolated from 88 Mexican children with diarrhea was screened by PCR. Among 82 ETEC isolates found, 30 (36.5%) were lngA PCR-positive. Twenty-seven percent of the children shed ETEC that possessed lngA. In parallel with DNA probes or PCR protocols to detect enterotoxin genes, the lngA PCR method may prove useful for detection of ETEC harboring type IV pilus genes in epidemiological studies. PMID- 10790097 TI - A PCR-colorimetric microwell plate hybridization assay for detection of Mycobacterium tuberculosis and M. avium from culture samples and Ziehl-Neelsen positive smears. AB - Differentiation between Mycobacterium tuberculosis and M. avium is essential for the treatment of mycobacterial infections. We have developed an easy and rapid detection assay for the diagnosis of mycobacterial diseases. This is a PCR hybridization assay based on selective amplification of a 16S rRNA gene sequence using pan-Mycobacterium primers followed by hybridization of the amplification products to biotinylated M. tuberculosis and M. avium-specific probes. A total of 55 mycobacterial isolates were tested. For all isolates, results concordant with those of conventional identification methods were obtained. Moreover, we developed a method for extraction of DNA from Ziehl-Neelsen-positive smears which allows the recovery of intact target DNA in our PCR-hybridization assay. Our method was able to confirm all culture results for 59 Ziehl-Neelsen-positive smears from clinical specimens (35 sputum, 11 lymph node biopsy, 6 stool, 4 pus, 2 urine, and 1 pericardial fluid specimens). These data suggest that our PCR hybridization assay, which is simple to perform and less expensive than commercial probe methods, may be suitable for the identification of M. tuberculosis and M. avium. It could become a valuable alternative approach for the diagnosis of mycobacterial infections when applied directly to DNA extracted from Ziehl-Neelsen-positive smears as well. PMID- 10790098 TI - Immunoblot analysis of humoral immune response to Helicobacter pylori in children with and without duodenal ulcer. AB - Several studies have demonstrated that enzyme-linked immunosorbent assay is not a sensitive and specific method to diagnose Helicobacter pylori infection in children, especially in the younger ones. Since serum immune response can also be determined by immunoblotting and it permits the detection of antibodies to virulence factors such as CagA and VacA, we evaluated the accuracy of a commercial immunoblotting test to diagnose H. pylori infection and to assess the humoral immune response to different H. pylori antigens in 122 children who underwent upper gastrointestinal endoscopy. The presence of H. pylori was determined in antral biopsy specimens by culture, preformed urease test, and histological analysis. H. pylori was identified by microbiological and histopathological methods in 66 children (including all of the 21 who had duodenal ulcer). Antibodies to H. pylori were detected in 63 infected children and in 8 noninfected ones. The sensitivity, specificity, and positive and negative predictive values of the immunoblotting test were 95.5, 85.7, 88.7, and 94.1%, respectively. The number of immunoreactive bands increased with age (P = 0.003), and the bands of 35 kDa (P = 0.013); 89 kDa, the VacA antigen (P = 0.001); and 116 kDa, the CagA antigen (P = 0.00004) were more frequently observed in older children. The frequency of the bands of 89 kDa (P = 0.001) and 116 kDa (P = 0.03) was higher in children with duodenal ulcer than in H. pylori-positive children without the disease. In conclusion, the immunoblotting test appears to be useful for the diagnosis of H. pylori infection in children, even in the younger ones. PMID- 10790099 TI - Evaluation of the VITEK 2 system for rapid identification of yeasts and yeast like organisms. AB - The new VITEK 2 system is a fully automated system dedicated to the identification and susceptibility testing of microorganisms. In conjunction with the VITEK ID-YST card the VITEK 2 system allows the identification of clinically important yeasts and yeast-like organisms in 15 h due to a sensitive fluorescence based technology. The ID-YST card consists of 47 biochemical reactions. The database comprises 51 taxa, including newly described species. In this study we evaluated the reliability of the VITEK ID-YST card for the identification of yeasts and yeast-like organisms encountered in a clinical microbiology laboratory. A total of 241 strains representing 21 species were studied. The strains were isolated from clinical samples within a period of 60 days prior to the identification. The tests were performed using 24-h to 55-h subcultures on Sabouraud-gentamicin-chloramphenicol agar. Each strain was tested in parallel using the ID 32C strip as a comparison method combined with microscopic morphology and an agglutination test for C. krusei. Overall, 222 strains (92.1%) were unequivocally identified including 11 isolates (4.6%) identified with low discrimination resolved by simple additional tests. Ten strains (4. 1%) for which results were given with low discrimination could not be unequivocally identified with supplemental tests, 4 strains (1. 7%) were misidentified and 5 strains (2.1%) could not be identified. In conclusion, we found that the VITEK 2 system is a rapid and accurate method for the identification of medically important yeasts and yeast-like organisms. PMID- 10790100 TI - Sequence diversity of the Escherichia coli H7 fliC genes: implication for a DNA based typing scheme for E. coli O157:H7. AB - Flagellar (H) antigens are mostly encoded by genes at the fliC locus in E. coli. We have sequenced 11 H7 fliC genes from Escherichia coli strains that belong to seven O serotypes. These sequences, together with those of nine other H7 fliC genes (from strains of three different O serotypes) sequenced recently (S. D. Reid, R. K. Selander, and T. S. Whittam, J. Bacteriol. 181:153-160, 1999), include 10 different sequences. The differences between these 10 sequences range from 0.06 to 3.12%. By comparison with other E. coli flagellin genes, we have identified primer length sequences specific for H7 genes in general and others specific for H7 genes of O157 and O55 strains: the specificity was confirmed by PCR testing the type strains for all 53 E. coli H types. We have previously identified genes specific for the E. coli O157 antigen, and use of the combination of O157- and H7-specific primers allows the sensitive and rapid detection of O157:H7 E. coli strains, which cause the majority of hemorrhagic colitis cases. PMID- 10790101 TI - Occurrence and detection of AmpC beta-lactamases among Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis isolates at a veterans medical center. AB - AmpC beta-lactamases are cephalosporinases that confer resistance to a wide variety of beta-lactam drugs and that may thereby create serious therapeutic problems. Although reported with increasing frequency, the true rate of occurrence of AmpC beta-lactamases in Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis remains unknown. We tested a total of 1,286 consecutive, nonrepeat isolates of these three species and found that, overall, 45 (3.5%) yielded a cefoxitin zone diameter less than 18 mm (screen positive) and that 16 (1.2%) demonstrated AmpC bands by isoelectric focusing. Based on the species, of 683 E. coli, 371 K. pneumoniae, and 232 P. mirabilis isolates tested, 13 (1.9%), 28 (7.6%), and 4 (1.7%), respectively, demonstrated decreased zone diameters and 11 (1.6%), 4 (1.1%), and 1 (0.4%), respectively, demonstrated AmpC bands. Cefoxitin resistance was transferred for all but 8 (E. coli) of the 16 AmpC producers. We also describe a three-dimensional extract test, which was used to detect phenotypically isolates that harbor AmpC beta-lactamase. Of the 45 cefoxitin-resistant isolates, the three-dimensional extract test accurately identified all 16 AmpC producers and 28 of 29 (97%) isolates as non-AmpC producers. Interestingly, most (86%) isolates in the latter group were K. pneumoniae isolates. These data confirm that, at our institution, E. coli, K. pneumoniae, and P. mirabilis harbor plasmid-mediated AmpC enzymes. PMID- 10790102 TI - Oxygen and carbon dioxide regulation of toxic shock syndrome toxin 1 production by Staphylococcus aureus MN8. AB - The production of toxic shock syndrome toxin 1 (TSST-1) by Staphylococcus aureus MN8 exposed to a range of oxygen concentrations (0 to 21% [vol/vol]) was examined in batch and thin-film cultures. The response of S. aureus to this range of oxygen concentrations was studied in the absence and in the presence of 7% (vol/vol) carbon dioxide. In the absence of carbon dioxide, TSST-1 production in batch cultures increased from negligible levels in the presence of oxygen concentrations of 1% or less to 500 ng/ml in the presence of 2% oxygen and then decreased to 70 ng/ml or less in the presence of oxygen concentrations of 6% and higher. In the presence of carbon dioxide, however, toxin production increased from negligible levels in the presence of 1% oxygen to 1,900 ng/ml in the presence of 21% oxygen. In thin-film cultures, TSST-1 production increased from nearly undetectable levels under anaerobic conditions to 1 and 10 microg/ml under 21% oxygen in the absence and presence of carbon dioxide, respectively. This study demonstrates the controlling effects of both oxygen and carbon dioxide on TSST-1 production. PMID- 10790104 TI - Phylogeny of Porphyromonas gingivalis by ribosomal intergenic spacer region analysis. AB - Periodontitis has been associated with the presence of Porphyromonas gingivalis, and previous studies have shown phenotypic differences in the pathogenicities of strains of P. gingivalis. An accurate and comprehensive phylogeny of strains of P. gingivalis would be useful in determining if there is an evolutionary basis to pathogenicity in this species. Previous phylogenies of P. gingivalis strains based on random amplified polymorphic DNA (RAPD) analysis and multilocus enzyme electrophoresis (MLEE) show little agreement. While the 16S ribosomal gene is the standard for phylogenetic reconstruction among bacterial species, it is insufficiently variable for this purpose. In the present study, the phylogeny of P. gingivalis was constructed on the basis of the sequence of the most variable region of the ribosomal operon, the intergenic spacer region (ISR). Heteroduplex analysis of the ISR has been used to study the variability of P. gingivalis strains in periodontitis. In the present study, typing by heteroduplex analysis was compared to ISR sequence-based phylogeny and close agreement was observed. The two strains of P. gingivalis whose heteroduplex types are strongly associated with periodontitis were found to be closely related and were well separated from strains whose heteroduplex types are less strongly associated with disease, suggesting a relationship between pathogenicity and phylogeny. PMID- 10790103 TI - Distribution of rotavirus VP4 genotypes and VP7 serotypes among nonhospitalized and hospitalized patients with gastroenteritis and patients with nosocomially acquired gastroenteritis in Austria. AB - To assess the potential benefits of a reassortant tetravalent rotavirus vaccine, we investigated stool specimens from children in three different groups by reverse transcription-PCR (RT-PCR) for rotavirus G and P types: (i) children not hospitalized with community-acquired rotavirus-acute gastroenteritis (RV-AGE), (ii) children hospitalized for RV-AGE, and (iii) children with nosocomially acquired RV-AGE. From a total of 553 samples investigated, 335 were positive by enzyme-linked immunosorbent assay, of which 294 (88%) were positive by RT-PCR. Among the RT-PCR-positive samples, the predominant types were G1P[8] (84%), followed by G4P[8] (9%) and G3P[8] (2%). No differences between the three groups were observed, suggesting that community vaccination will diminish the most cost relevant cases of hospitalizations and nosocomial infections. PMID- 10790105 TI - Helicobacter mesocricetorum sp. nov., A novel Helicobacter isolated from the feces of Syrian hamsters. AB - A spiral-shaped bacterium with bipolar, single, nonsheathed flagella was isolated from the feces of Syrian hamsters. The bacterium grew as a thin spreading film at 37 degrees C under microaerobic conditions, did not hydrolyze urea, was positive for catalase and alkaline phosphatase, reduced nitrate to nitrite, did not hydrolyze hippurate, and was sensitive to nalidixic acid but resistant to cephalothin. Sequence analysis of the 16S rRNA gene and biochemical and phenotypic criteria indicate that the novel bacterium is a helicobacter. The novel bacterium is most closely related to the recently described mouse enteric helicobacter, Helicobacter rodentium. This is the first urease-negative Helicobacter species with nonsheathed flagella isolated from feces of asymptomatic Syrian hamsters. We propose to name this novel helicobacter Helicobacter mesocricetorum. The type strain is MU 97-1514 (GenBank accession number AF072471). PMID- 10790106 TI - Comparison of agar diffusion methodologies for antimicrobial susceptibility testing of Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - Pseudomonas aeruginosa is the most common pathogen infecting the lungs of patients with cystic fibrosis (CF). Improved antimicrobial chemotherapy has significantly increased the life expectancy of these patients. However, accurate susceptibility testing of P. aeruginosa isolates from CF sputum may be difficult because the organisms are often mucoid and slow growing. This study of 597 CF isolates of P. aeruginosa examined the correlation of disk diffusion and Etest (AB BIODISK, Solna, Sweden) results with a reference broth microdilution method. The rates of interpretive errors for 12 commonly used antipseudomonal antimicrobials were determined. The disk diffusion method correlated well (zone diameter versus MIC) for all of the agents tested. However, for mucoid isolates, correlation coefficients (r values) for piperacillin, piperacillin-tazobactam, and meropenem were <0.80. The Etest correlation with reference broth microdilution results (MIC versus MIC) was acceptable for all of the agents tested, for both mucoid and nonmucoid isolates. Category interpretation errors were similar for the disk diffusion and Etest methods with 0.4 and 0.1%, respectively, very major errors (false susceptibility) and 1.1 and 2.2% major errors (false resistance). Overall, both agar diffusion methods appear to be broadly acceptable for routine clinical use in susceptibility testing of CF isolates of P. aeruginosa. PMID- 10790107 TI - Standardization of immunoglobulin M capture enzyme-linked immunosorbent assays for routine diagnosis of arboviral infections. AB - Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) is a rapid and versatile diagnostic method that readily permits the combination of multiple assays. Test consolidation is especially important for arthropod borne viruses (arboviruses) which belong to at least three virus families: the Togaviridae, Flaviviridae, and Bunyaviridae. Using prototype viruses from each of these families and a panel of well-characterized human sera, we have evaluated and standardized a combined MAC-ELISA capable of identifying virus infections caused by members of each virus family. Furthermore, by grouping antigens geographically and utilizing known serological cross-reactivities, we have reduced the number of antigens necessary for testing, while maintaining adequate detection sensitivity. We have determined that a 1:400 serum dilution is most appropriate for screening antiviral antibody, using a positive-to-negative ratio of >/=2.0 as a positive cutoff value. With a blind-coded human serum panel, this combined MAC-ELISA was shown to have test sensitivity and specificity that correlated well with those of other serological techniques. PMID- 10790108 TI - Detection of anti-arboviral immunoglobulin G by using a monoclonal antibody-based capture enzyme-linked immunosorbent assay. AB - Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from the Alphavirus, Flavivirus, and Bunyavirus genera. Tests were optimized and standardized so that maximum homology could be maintained among working parameters for the different viral agents, enabling a wide range of viruses to be easily tested for at one time. MAbs were screened for suitability as capture vehicles for antigens from the three genera. The final test configuration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capture the specific inactivated viral antigens. Serum IgG was detected by using alkaline phosphatase conjugated anti-human IgG (Fc portion). A dilution of 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false-positive results. IgG ELISA results correlated with those of the standard plaque-reduction neutralization assays. As expected, some test cross-reactivity was encountered within the individual genera, and tests were interpreted within the context of these reactions. The tests were standardized for laboratory diagnosis of arboviral infections, with the intent that they be used in tandem with the corresponding IgM antibody capture ELISAs. PMID- 10790109 TI - Transfer of erythromycin resistance from poultry to human clinical strains of Staphylococcus aureus. AB - The transfer of ermA and ermC genes, the two most common resistance determinants of erythromycin resistance, was studied with Luria-Bertani broth in the absence of additional Ca(2+) or Mg(2+) ions. Fifteen human and five poultry isolates of Staphylococcus aureus, which were resistant to erythromycin but carried different genetic markers for erythromycin resistance, were used for conjugation. Since both the donors (Amp(s)-Tet(r)) and recipients (Amp(r)-Tet(s)) were resistant to erythromycin, the transconjugants were initially picked up as ampicillin- and tetracycline-resistant colonies. The resistance transfer mechanisms of the chromosomally located erythromycin rRNA methylase gene ermA and the plasmid-borne ermC gene were monitored by a multiplex PCR and gene-specific internal probing assay. Four groups of transconjugants, based upon the transfer of the ermA and/or ermC gene, were distinguished from each other by the use of this method. Selective antibiotic screening revealed only one type of transconjugant that was resistant to ampicillin and tetracycline. A high frequency of transfer (4.5 x 10( 3)) was observed in all of the 23 transconjugants obtained, and the direction of tetracycline and erythromycin resistance marker transfer was determined to be from poultry to clinical isolates. The transfers of the ermA and ermC genes were via transposition and transformation, respectively. PMID- 10790110 TI - Rapid phenotypic characterization method for herpes simplex virus and Varicella Zoster virus thymidine kinases to screen for acyclovir-resistant viral infection. AB - A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella zoster virus (VZV) thymidine kinase (TK) genes was developed for monitoring acyclovir-resistant viruses. This method determines the biochemical phenotype of the TK polypeptide, which is synthesized in vitro from viral DNA using a procedure as follows. The TK gene of each sample virus strain is amplified and isolated under the control of a T7 promoter by PCR. The PCR products are transcribed with T7 RNA polymerase and translated in a rabbit reticulocyte lysate. Using this method, enzymatic characteristics and the size of the TK polypeptides encoding HSV and VZV DNA were defined in less than 2 days without virus isolation. The assay should be a powerful tool in monitoring drug-resistant viruses, especially in cases in which virus isolation is difficult. PMID- 10790112 TI - Development of a fluorescence polarization-based diagnostic assay for equine infectious anemia virus. AB - The control of equine infectious anemia virus (EIAV) infections of horses has been over the past 20 years based primarily on the identification and elimination of seropositive horses, predominantly by a standardized agar gel immunodiffusion (AGID) assay in centralized reference laboratories. This screening for EIAV seropositive horses has been to date hindered by the lack of a rapid diagnostic format that can be easily employed in the field. We describe here the development of a rapid solution-phase assay for the presence of serum antibodies to EIAV based on fluorescence polarization (FP) (patent pending). Peptides derived from antigenic regions of EIAV core and envelope proteins were initially screened for their utility as probes in an FP assay to select the best peptide antigen candidates. The FP assay was optimized to detect the presence of EIAV-specific antibodies by a change in the FP of a fluorescein-labeled immunoreactive peptide diagnostic antigen. The most sensitive and specific peptide probe was a peptide corresponding to the immunodominant region of the EIAV transmembrane protein, gp45. This probe was tested for its reactivity in the optimized FP assay with 151 AGID-positive horse sera and 106 AGID-negative serum samples. The results of these studies demonstrated that the FP assay reactivity correlated with reported AGID results in 106 of 106 negative serum samples (100% specificity) and in 135 of 151 positive serum samples (89.4% sensitivity). The FP assay was also found to have a very low background reactivity and to readily detect antibodies produced early in infection (90.0, >70.0, and >80.0%, respectively, for all four assays. The mean times to first HCMV detection after bone marrow transplantation were 37.7 +/- 15.4 days for detection of IE mRNA by NASBA, 39.6 +/- 15.6 days for quantitation of antigenemia, 40.9 +/- 15.2 days for quantitation of DNAemia, and 43.7 +/- 16.3 or 43.7 +/- 17.5 days for quantitation of viremia and detection of pp67 mRNA by NASBA, respectively. On the whole, 31 BMT recipients received preemptive therapy by using confirmed antigenemia positivity as a cutoff, while 35 patients could have been treated by using NASBA positivity as a cutoff and 31 could have been treated by using quantitation of DNAemia as a cutoff. In single patients, IE mRNA was detected in every episode of active HCMV infection, mostly preceding and sometimes accompanying antigenemia and DNAemia, whereas pp67 mRNA was detected only concomitantly with the highest peaks of infection. HCMV IE mRNA detection may represent a useful parameter for initiation of preemptive therapy in BMT recipients. PMID- 10790114 TI - Performance of the ImmunoCard STAT! E. coli O157:H7 test for detection of Escherichia coli O157:H7 in stools. AB - ImmunoCard STAT! E. coli O157:H7 (Meridian Diagnostics, Inc., Cincinnati, Ohio) is a novel rapid (10-min) test for the presence of Escherichia coli O157:H7 in stools. The test may be performed either directly on stool specimens or on an overnight broth culture of stool. In a multicenter prospective study, 14 of 14 specimens positive by culture for E. coli O157:H7 were positive by the ImmunoCard STAT! O157:H7 test, and there were no false positives from 263 culture-negative specimens. In a retrospective study, the test was positive in 339 (81%) of 417 stored culture-positive specimens and the specificity was 95% (98 of 103 specimens). No false positives were associated with alternate stool pathogens. The ImmunoCard STAT! O157:H7 test has high sensitivity and specificity. PMID- 10790113 TI - Genetic relatedness of Salmonella isolates from nondomestic birds in Southeastern United States. AB - Salmonella infections have been implicated in large-scale die-offs of wild birds in the United States. Although we know quite a bit about the epidemiology of Salmonella infection among domestic fowl, we know little about the incidence, epidemiology, and genetic relatedness of salmonellae in nondomestic birds. To gain further insight into salmonellae in these hosts, 22 Salmonella isolates from diseased nondomestic birds were screened for the presence of virulence and antibiotic resistance-associated genes and compared genetically using pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA analysis. Of the 22 Salmonella isolates examined, 15 were positive for the invasion gene invA and the virulence plasmid-associated genes spvC and pef. Most (15 of 22) were generally sensitive to antibiotics. However, two Salmonella isolates from pet birds were identified as Salmonella enterica serovar Typhimurium DT104. Despite the general susceptibility of these Salmonella isolates to most antimicrobial agents, antibiotic resistance-associated genes intI1, merA, and aadA1 were identified in a number of these isolates. Five distinct XbaI and nine distinct BlnI DNA patterns were observed for the 22 Salmonella isolates typed by PFGE. PFGE analysis determined that Salmonella isolates from passerines in Georgia and Wyoming were genetically related. PMID- 10790115 TI - Molecular differentiation of seven Malassezia species. AB - A system based on PCR and restriction endonuclease analysis was developed to distinguish the seven currently recognized Malassezia species. Seventy-eight strains, including authentic culture collection strains and routine clinical isolates, were investigated for variation in the ribosomal DNA repeat units. Two genomic regions, namely, the large subunit of the ribosomal gene and the internal transcribed spacer (ITS) region, were amplified by PCR, and products were digested with restriction endonucleases. The patterns generated were useful in identification of five out of seven Malassezia species. M. sympodialis was readily distinguishable in that its ITS region yielded a 700-bp amplified fragment, whereas the other six species yielded an 800-bp fragment. M. globosa and M. restricta were very similar in the regions studied and could be distinguished only by performing a hot start-touchdown PCR on primers for the beta-tubulin gene. Primers based on the conserved areas of the Candida cylindracea lipase gene, which were used in an attempt to amplify Malassezia lipases, yielded an amplification product after annealing at 55 degrees C only with M. pachydermatis. This specific amplification may facilitate the rapid identification of this organism. PMID- 10790116 TI - Distinguishing species of the Burkholderia cepacia complex and Burkholderia gladioli by automated ribotyping. AB - Several species belonging to the genus Burkholderia are clinically relevant, opportunistic pathogens that inhabit major environmental reservoirs. Consequently, the availability of means for adequate identification and epidemiological characterization of individual environmental or clinical isolates is mandatory. In the present communication we describe the use of the Riboprinter microbial characterization system (Qualicon, Warwick, United Kingdom) for automated ribotyping of 104 strains of Burkholderia species from diverse sources, including several publicly accessible collections. The main outcome of this analysis was that all strains were typeable and that strains of Burkholderia gladioli and of each species of the B. cepacia complex, including B. multivorans, B. stabilis, and B. vietnamiensis, were effectively discriminated. Furthermore, different ribotypes were discerned within each species. Ribotyping results were in general agreement with strain classification based on restriction fragment analysis of 16S ribosomal amplicons, but the resolution of ribotyping was much higher. This enabled automated molecular typing below the species level. Cluster analysis of the patterns obtained by ribotyping (riboprints) showed that within B. gladioli, B. multivorans, and B. cepacia genomovar VI, the different riboprints identified always clustered together. Riboprints of B. cepacia genomovars I and III, B. stabilis, and B. vietnamiensis did not show distinct clustering but rather exhibited the formation of loose assemblages within which several smaller, genomovar-specific clusters were delineated. Therefore, ribotyping proved useful for genomovar identification. Analysis of serial isolates from individual patients demonstrated that infection with a single ribotype had occurred, despite minor genetic differences that were detected by pulsed-field gel electrophoresis of DNA macrorestriction fragments. The automated approach allows very rapid and reliable identification and epidemiological characterization of strains and generates an easily manageable database suited for expansion with information on additional bacterial isolates. PMID- 10790117 TI - Effects of bovine herpesvirus type 1 infection in calves with maternal antibodies on immune response and virus latency. AB - The presence of maternally derived antibodies can interfere with the development of an active antibody response to antigen. Infection of seven passively immunized young calves with a virulent strain of bovine herpesvirus type 1 (BHV-1) was performed to determine whether they could become seronegative after the disappearance of maternal antibodies while latently infected with BHV-1. Four uninfected calves were controls. All calves were monitored serologically for 13 to 18 months. In addition, the development of a cell-mediated immune response was assessed by an in vitro antigen-specific gamma interferon (IFN-gamma) production assay. All calves had positive IFN-gamma responses as early as 7 days until at least 10 weeks after infection. However, no antibody rise was observed after infection in the three calves with the highest titers of maternal antibodies. One of the three became seronegative by virus neutralization test at 7 months of age like the control animals. This calf presented negative IFN-gamma results at the same time and was classified seronegative by enzyme-linked immunosorbent assay at around 10 months of age. This calf was latently infected, as proven by virus reexcretion after dexamethasone treatment at the end of the experiment. In conclusion, this study demonstrated that BHV-1-seronegative latent carriers can be obtained experimentally. In addition, the IFN-gamma assay was able to discriminate calves possessing only passively acquired antibodies from those latently infected by BHV-1, but it could not detect seronegative latent carriers. The failure to easily detect such animals presents an epidemiological threat for the control of BHV-1 infection. PMID- 10790118 TI - Direct molecular typing of Borrelia burgdorferi sensu lato species in synovial samples from patients with lyme arthritis. AB - Since Lyme arthritis was first described in the United States, it has now been reported in many countries of Europe. However, very few strains of the causative bacterium, Borrelia burgdorferi, have been isolated from synovial samples. For this reason, different molecular direct typing methods were developed recently to assess which species could be involved in Lyme arthritis in Europe. We developed a simple oligonucleotide typing method with PCR fragments from the flagellin gene of B. burgdorferi sensu lato, which is able to differentiate seven different Borrelia species. Among 10 consecutive PCR-positive patients with Lyme arthritis from the northeastern France, two species were identified in synovial samples: B. burgdorferi sensu stricto in 9 cases and B. garinii in 1 case. Conversely, all B. burgdorferi sensu lato species detected in 10 consecutive PCR-positive biopsies from a second set of patients with erythema migrans from the same geographical area were identified as either B. afzelii or B. garinii (P < 0.001). These results indicate that B. burgdorferi sensu stricto is the principal but not the only Borrelia species involved in Lyme arthritis in northeastern France. PMID- 10790119 TI - Technical assessment of the affymetrix yeast expression GeneChip YE6100 platform in a heterologous model of genes that confer resistance to antimalarial drugs in yeast. AB - The advent of high-density gene array technology has revolutionized approaches to drug design, development, and characterization. At the laboratory level, the efficient, consistent, and dependable exploitation of this complex technology requires the stringent standardization of protocols and data analysis platforms. The Affymetrix YE6100 expression GeneChip platform was evaluated for its performance in the analysis of both global (6,000 yeast genes) and targeted (three pleiotropic multidrug resistance genes of the ATP binding cassette transporter family) gene expression in a heterologous yeast model system in the presence and absence of the antimalarial drug chloroquine. Critical to the generation of consistent data from this platform are issues involving the preparation of the specimen, use of appropriate controls, accurate assessment of experiment variance, strict adherence to optimized enzymatic and hybridization protocols, and use of sophisticated bioinformatics tools for data analysis. PMID- 10790120 TI - Distribution and molecular characterization of Porphyromonas gingivalis carrying a new type of fimA gene. AB - Fimbriae of Porphyromonas gingivalis are filamentous appendages on the cell surface and are thought to be one of the virulence factors. The fimA gene encoding the subunit protein of fimbriae, fimbrillin (FimA), was classified into four typeable variants (types I to IV). We previously examined the distribution of P. gingivalis in terms of fimA genotypes in periodontitis patients using a fimA type-specific PCR assay. However, some patients harbored P. gingivalis with untypeable fimA. In this study, we have cloned a new type (type V) of fimA from dental plaque samples. P. gingivalis with type V fimA was isolated from dental plaque of a periodontitis patient, and the isolate was named HNA-99. The deduced amino acid sequences were compared with those of type I P. gingivalis ATCC 33277, type II strain HW24D1, type III strain 6/26, and type IV strain HG564, and the homologies were found to be 45, 44, 43, and 55%, respectively. Southern blot analysis showed that the clinical isolate HNA-99 possessed P. gingivalis-specific genes sod and kgp. However, in terms of serological specificities, type V FimA showed a difference from other types of FimA. In addition, type V P. gingivalis bacteria were detected in 16.4% (12 of 73) of the P. gingivalis-positive patients with periodontitis by PCR assay using specific primers. Thus, a new type of fimA gene is now established, and the fimA genotyping could be useful in determining the disease-associated genotypes of P. gingivalis involved in the development of adult periodontitis. PMID- 10790121 TI - Evaluation of the LiPA MYCOBACTERIA assay for identification of mycobacterial species from BACTEC 12B bottles. AB - The LiPA MYCOBACTERIA (Innogenetics NV, Ghent, Belgium) assay was used to identify mycobacterial isolates using culture fluid from positive BACTEC 12B bottles. The LiPA method involves reverse hybridization of a biotinylated mycobacterial PCR fragment, a 16 to 23S rRNA spacer region, to oligonucleotide probes arranged in lines on a membrane strip, with detection via biotin streptavidin coupling by a colorimetric system. This system identifies Mycobacterium species and differentiates M. tuberculosis complex, M. avium-M. intracellulare complex, and the following mycobacterial species: M. avium, M. intracellulare, M. kansasii, M. chelonae group, M. gordonae, M. xenopi, and M. scrofulaceum. The mycobacteria were identified in the laboratory by a series of tests, including the Roche AMPLICOR Mycobacterium tuberculosis (MTB) test, the Gen-Probe ACCUPROBE, and a PCR-restriction fragment length polymorphism (PCR RFLP) analysis of the 65-kDa heat shock protein gene. The LiPA MYCOBACTERIA assay detected 60 mycobacterium isolates from 59 patients. There was complete agreement between LiPA and the laboratory identification tests for 26 M. tuberculosis complex, 9 M. avium, 3 M. intracellulare complex, 3 M. kansasii, 4 M. gordonae, and 5 M. chelonae group (all were M. abscessus) isolates. Three patient samples were LiPA positive for M. avium-M. intracellulare complex, and all were identified as M. intracellulare by the PCR-RFLP analysis. Seven additional mycobacterial species were LiPA positive for Mycobacterium spp. (six were M. fortuitum, and one was M. szulgai). The LiPA MYCOBACTERIA assay was easy to perform, and the interpretation of the positive bands was clear-cut. Following PCR amplification and gel electrophoresis, the LiPA assay was completed within 3 h. PMID- 10790123 TI - Coinfection with multiple TT virus strains belonging to different genotypes is a common event in healthy Brazilian adults. AB - Testing of the DNA of TT virus (TTV) was done with serum samples obtained from 191 persons working in a public hospital of the city of Rio de Janeiro, Brazil. TTV DNA was detected by PCR in the sera of 125 (65.4%) individuals. PCR products were cloned, and sequences with a length of 159 bases surrounding the TATA signal region were determined for 100 clones derived from 31 individuals. One clone from each of 23 subjects was sequenced, while 7 to 19 clones from eight individuals were sequenced. None of the sera contained a viral sequence identical to that of any other individual. Phylogenetic analysis revealed the existence of a divergent TTV genotype possessing a single-base deletion at position 140. Among the eight persons for whom various sequences were analyzed, six were coinfected with between two and seven TTV strains belonging to different genotypes. The results suggest that coinfection with multiple TTV strains belonging to different genotypes is a common event in healthy Brazilian adults. PMID- 10790122 TI - Detection of human cytomegalovirus pp67 late gene transcripts in cerebrospinal fluid of human immunodeficiency virus type 1-infected patients by nucleic acid sequence-based amplification. AB - This study examined the clinical correlation between the presence of human cytomegalovirus (HCMV) pp67 mRNA in cerebrospinal fluid (CSF) and active HCMV central nervous system (CNS) disease in patients with human immunodeficiency virus type 1 (HIV-1). In total, 76 CSF specimens collected from 65 HIV-1-positive patients diagnosed with HCMV CNS disease, other non-HCMV-related CNS diseases, or no CNS disease were tested for the presence of HCMV pp67 mRNA using the NucliSens cytomegalovirus (CMV) pp67 assay (Organon Teknika, Durham, N.C.). The results were compared to those of a nested PCR for the detection of HCMV glycoprotein B DNA and to those obtained by viral culture (54 samples). CSF specimens collected from patients without HCMV CNS disease yielded the following results: pp67 assay negative, 62 of 62 specimens; culture negative, 41 of 41 specimens; and PCR negative, 56 of 62 specimens (6 specimens were positive). CSF specimens collected from patients with HCMV CNS disease yielded the following results: pp67 assay positive, 9 of 13 specimens; PCR positive, 13 of 13 specimens; and culture positive, 2 of 13 specimens. After resolution of the discordant results, the following positive and negative predictive values (PPV and NPV, respectively) for the diagnosis of HCMV CNS disease were determined. The PPV for PCR, pp67 assay, and culture were 68.4, 100, and 100%, respectively, and the NPV for PCR, pp67 assay, and culture were 100, 97.0, and 82. 7%, respectively. The sensitivities for DNA PCR, pp67 assay, and culture for the detection of HCMV were 100, 84.6, and 18%, respectively, and the clinical specificities were 90.5, 100, and 100%, respectively. This study indicates that the detection of HCMV pp67 mRNA in CSF has good correlation with active HCMV CNS disease, whereas CSF culture is insensitive and qualitative DNA PCR may detect latent nonreplicating virus in CSF from patients without HCMV CNS disease. PMID- 10790124 TI - Application of pulsed-field gel electrophoresis and binary typing as tools in veterinary clinical microbiology and molecular epidemiologic analysis of bovine and human Staphylococcus aureus isolates. AB - Thirty-eight bovine mammary Staphylococcus aureus isolates from diverse clinical, temporal, and geographical origins were genotyped by pulsed-field gel electrophoresis (PFGE) after SmaI digestion of prokaryotic DNA and by means of binary typing using 15 strain-specific DNA probes. Seven pulsed-field types and four subtypes were identified, as were 16 binary types. Concordant delineation of genetic relatedness was documented by both techniques, yet based on practical and epidemiological considerations, binary typing was the preferable method. Genotypes of bovine isolates were compared to 55 previously characterized human S. aureus isolates through cluster analysis of binary types. Genetic clusters containing strains of both human and bovine origin were found, but bacterial genotypes were predominantly associated with a single host species. Binary typing proved an excellent tool for comparison of S. aureus strains, including methicillin-resistant S. aureus, derived from different host species and from different databases. For 28 bovine S. aureus isolates, detailed clinical observations in vivo were compared to strain typing results in vitro. Associations were found between distinct genotypes and severity of disease, suggesting strain-specific bacterial virulence. Circumstantial evidence furthermore supports strain-specific routes of bacterial dissemination. We conclude that PFGE and binary typing can be successfully applied for genetic analysis of S. aureus isolates from bovine mammary secretions. Binary typing in particular is a robust and simple method and promises to become a powerful tool for strain characterization, for resolution of clonal relationships of bacteria within and between host species, and for identification of sources and transmission routes of bovine S. aureus. PMID- 10790125 TI - Computer-assisted analysis and epidemiological value of genotyping methods for Campylobacter jejuni and Campylobacter coli. AB - For epidemiological tracing of the thermotolerant Campylobacter species C. jejuni and C. coli, reliable and highly discriminatory typing techniques are necessary. In this study the genotyping techniques of flagellin typing (flaA typing), pulsed field gel electrophoresis (PFGE), automated ribotyping, and amplified fragment length polymorphism (AFLP) fingerprinting were compared. The following aspects were compared: computer-assisted analysis, discriminatory power, and use for epidemiological typing of campylobacters. A set of 50 campylobacter poultry isolates from The Netherlands and neighboring countries was analyzed. Computer assisted analysis made cluster analysis possible and eased the designation of different genotypes. AFLP fingerprinting was the most discriminatory technique, identifying 41 distinct genotypes, while PFGE identified 38 different types, flaA typing discriminated 31 different types, and ribotyping discriminated 26 different types. Furthermore, AFLP analysis was the most suitable method for computer-assisted data analysis. In some cases combining the results of AFLP fingerprinting, PFGE, and flaA typing increased our ability to differentiate strains that appeared genetically related. We conclude that AFLP is a highly discriminatory typing method and well suited for computer-assisted data analysis; however, for optimal typing of campylobacters, a combination of multiple typing methods is needed. PMID- 10790126 TI - Development and evaluation of a molecular viability assay for Pneumocystis carinii. AB - Despite recent declines in incidence, Pneumocystis carinii pneumonia (PCP) remains the most commonly occurring opportunistic illness among persons with AIDS in the United States. While P. carinii DNA has been detected in patient respiratory specimens and in air samples collected from various indoor environments housing PCP patients, the viability of these organisms is unknown. For this reason, we have developed and evaluated a molecular viability assay for P. carinii. This method is based upon the detection of P. carinii mRNA by a reverse transcription-PCR that employs specific primers from a member of the heat shock protein 70 family. Under optimal assay conditions, these primers were capable of detecting as few as 100 viable trophozoites as determined by ethidium bromide staining, while no signal was obtained from 10(6) trophozoites killed by heat, desiccation, or UV radiation. This assay was also capable of distinguishing P. carinii from other common fungi present in the air. Therefore, this molecular viability assay may be useful in conjunction with standard bioaerosol collection devices and procedures for the detection of viable P. carinii collected from various indoor environments. It may also be useful in confirming the presence of viable trophozoites in respiratory specimens collected by noninvasive techniques from putatively infected individuals. PMID- 10790127 TI - Rapid identification of Yersinia enterocolitica in blood by the 5' nuclease PCR assay. AB - Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5' nuclease TaqMan PCR assay was developed to detect Y. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolitica cells, and total chromosomal DNA was extracted. When the TaqMan PCR assay was performed, as few as six bacteria spiked in 200 microliter of blood could be detected. The assay was specific and did not detect other Yersinia species. The TaqMan assay is easy to perform, takes 2 h, and has the potential for use in the rapid detection of Y. enterocolitica contamination in stored blood units. PMID- 10790128 TI - Evaluation of commercially available preservatives for laboratory detection of helminths and protozoa in human fecal specimens. AB - Formalin and mercuric chloride-based low-viscosity polyvinyl alcohol (LV-PVA) are widely used by most diagnostic parasitology laboratories for preservation of helminth eggs and protozoan cysts and trophozoites in fecal specimens. Concerns about the toxicity of formalin and the difficulty of disposal of LV-PVA are powerful incentives to use alternate preservatives. Such alternatives have been marketed by several companies and are often presented as one-vial, non-mercuric chloride fixatives that aim at performing the same role as formalin and PVA combined. We compared five, one-vial commercial preservatives, two from Meridian Diagnostics, Inc. (Ecofix and sodium acetate-acetic acid-formalin), and one each from Scientific Device Laboratories, Inc. (Parasafe), Alpha Tec Systems, Inc. (Proto-fix), and Streck Laboratories, Inc. (STF), with 10% formalin and LV-PVA. Fecal specimens obtained from patients in a Brazilian hospital were aliquoted within 12 h of collection into the seven preservatives mentioned above and were processed after 1 month at the Centers for Disease Control and Prevention. Direct and concentrated permanent smears as well as concentrates for 20 positive specimens (a total of 259 processed samples) were prepared, stained according to the manufacturers' instructions, examined, and graded. Positive specimens contained one or more parasites with stages consisting of eggs, larvae, cysts, and a few trophozoites of Giardia intestinalis. Criteria for assessment of the preservatives included the quality of the diagnostic characteristics of helminth eggs, protozoan cysts, and trophozoites, ease of use, and cost. Acceptable alternatives to formalin for wet preparations were found. Ecofix was found to be comparable to the traditional "gold standard" LV-PVA for the visualization of protozoa in permanent stained smears. This study suggests that more acceptable alternatives to the traditional formalin and LV-PVA exist. PMID- 10790129 TI - Acute cerebral phaeohyphomycosis due to Wangiella dermatitidis accompanied by cerebrospinal fluid eosinophilia. AB - We report a case of cerebral phaeohyphomycosis due to Wangiella dermaitidis in an immunocompetent adult man. His cerebrospinal fluid (CSF) showed pleocytosis with a high eosinophil count but without peripheral blood eosinophilia. The present case suggested that this black yeast-like fungus should be included when the causes of CSF eosinophilia are considered, even though it is an extremely rare pathogen. PMID- 10790130 TI - Macrorestriction fragment profiles reveal genetic variation of Cowdria ruminantium isolates. AB - Macrorestriction profile analysis by pulsed-field gel electrophoresis (PFGE) was used to distinguish between seven isolates of Cowdria ruminantium from geographically different areas. Characteristic profiles were generated for each isolate by using the restriction endonucleases KspI, SalI, and SmaI with chromosomal sizes ranging between 1,546 and 1,692 kb. Statistical analysis of the macrorestriction profiles indicated that all the isolates were distinct from each other; these data contribute to a better understanding of the epidemiology of this pathogen and may be exploited for the identification of genotype-specific DNA probes. PMID- 10790131 TI - Evidence from a nine-year birth cohort study in Japan of transmission pathways of Helicobacter pylori infection. AB - We examined the longitudinal changes of Helicobacter pylori infection within 46 families with children and 48 couples without children living in Japan. The study cohort was monitored from 1986 to 1994. H. pylori status was assessed by the presence of anti-H. pylori immunoglobulin G antibodies. At study entry, H. pylori prevalence in children with positive mothers was 23% versus 5% in children with negative mothers (odds ratio = 5.3; 95% confidence interval = 0.6 to 42.8). Seroconversion (rate of 1.5%/year) was evident only among children living with positive mothers and did not differ among adults living with or without children. These data strongly support the cluster phenomenon of H. pylori infection among families, the key role of the infected mothers in the transmission within families, and the importance of adult-child transmission and not vice versa. PMID- 10790132 TI - Prevalence of Cryptococcus neoformans var. neoformans (Serotype D) and Cryptococcus neoformans var. grubii (Serotype A) isolates in New York City. AB - Analysis of 40 New York City Cryptococcus neoformans isolates revealed that 39 were typeable, of which 85 and 12.5% were Cryptococcus neoformans var. grubii (serotype A) and Cryptococcus neoformans var. neoformans (serotype D), respectively. The prevalence of serotype D isolates in New York City appears to be significantly higher than indicated by previous studies of North American isolates. PMID- 10790133 TI - Evaluation of quantitative PCR and branched-chain DNA assay for detection of hepatitis B virus DNA in sera from hepatocellular carcinoma and liver transplant patients. AB - This study evaluated the applicability of quantitative PCR (Q-PCR) and branched chain DNA assays for detection of hepatitis B virus (HBV) DNA in sera. For 42 samples, the detection rates were 81 and 41%, respectively, with a correlation coefficient of 0.633. The Q-PCR is useful for early monitoring of HBV load in high-risk patients. PMID- 10790135 TI - Growth supplements for Helicobacter pylori. AB - The growth response of Helicobacter pylori in broth was determined in the presence of ferrous sulfate, sodium pyruvate, and mucin (porcine stomach). The addition of either ferrous sulfate and sodium pyruvate or mucin to brain heart infusion broth with 7% horse serum (BHI-HS) enhanced the growth of H. pylori. The best growth of strain NB2-1, which was the slowest growing of 10 H. pylori strains evaluated, occurred in the presence of 0.05% ferrous sulfate and 0.05% sodium pyruvate. The addition of 0.3% mucin to BHI-HS reduced the lag time of H. pylori by 48 h and enhanced the growth. On the basis of the results for 10 H. pylori strains, the combination of ferrous sulfate (0.025%), sodium pyruvate (0.025%), and mucin (0.15%) in BHI-HS counteracted the inhibitory effects of the antibiotics used in culture media for selective growth of H. pylori. Results suggest that these supplements may be useful for enhancement of the growth of H. pylori in enrichment media. PMID- 10790134 TI - Sensitive method for detection of human herpesviruses 6 and 7 in saliva collected in field studies. AB - To facilitate studies of the epidemiology and natural history of human herpesviruses 6 and 7 in infants, a practical method for collecting and quantifying the DNA of these viruses was developed. Saliva was collected using small strips of filter paper, and virus was detected using a real-time quantitative fluorescent-probe PCR assay. The sensitivity and specificity of this method even after prolonged drying of the specimens compared favorably to those of our traditional method of collecting and assaying saliva. PMID- 10790136 TI - Evaluation of the MB/BacT mycobacterium detection system for susceptibility testing of Mycobacterium tuberculosis. AB - The MB/BacT mycobacterium detection system was evaluated for its performance in the susceptibility testing of Mycobacterium tuberculosis. Eighty-three M. tuberculosis isolates were processed. Results for all isoniazid-, rifampin- and streptomycin-susceptible, isoniazid-resistant, and rifampin-resistant M. tuberculosis isolates with the MB/BacT system agreed 100% with those obtained by the agar proportion method. The agreements between the two methods for streptomycin- and ethambutol-resistant isolates were 96.4 and 90.4%, respectively. The susceptibility test results were obtained in 7 days, on average. These data demonstrate that the MB/BacT system is an accurate, nonradiometric method for rapid susceptibility testing of M. tuberculosis. PMID- 10790137 TI - Bartonella henselae infection as a cause of fever of unknown origin. AB - Fourteen of 41 patients (34%) with a serological diagnosis of Bartonella henselae infection were found to have prolonged fever or fever of unknown origin, suggesting that generalized systemic B. henselae infection is not rare in immunocompetent healthy individuals. PMID- 10790138 TI - Quantitation of cell-free and cell-associated Kaposi's sarcoma-associated herpesvirus DNA by real-time PCR. AB - A real-time PCR assay for quantitation of Kaposi's sarcoma-associated herpes virus (KSHV or human herpesvirus 8) DNA was evaluated. The linear dynamic range was 10 to 10(5) copies of KSHV DNA (r(2) > 0.99). The accuracy of DNA purification and quantitation was less than +/-0.4 log(10) copies for samples that contained from 10 to 10(5) copies of KSHV DNA. Cell-associated KSHV DNA was quantitated over a range of infected cell frequencies from 0. 1 to 10(-5), and cell-free KSHV DNA in plasma was quantitated over a range of 100 to 10(6) copies/ml. Real-time PCR provides a convenient method for quantitation of cell free and cell-associated KSHV DNA in laboratory and clinical specimens. PMID- 10790140 TI - A three-year study of Campylobacter jejuni genotypes in humans with domestically acquired infections and in chicken samples from the Helsinki area. AB - Campylobacter jejuni isolates from stool samples of patients with domestically acquired sporadic infections and from chicken from retail shops were studied during seasonal peaks from June to September over a 3-year period from 1996 to 1998. A large number of pulsed-field gel electrophoresis (PFGE) genotypes (a combined SmaI-SacII pattern) were identified each year. Certain genotypes persisted for the whole study period, and predominant genotypes represented 28 to 52% of the strains during a restricted period of time. The peak level of positive chicken samples was between July and August of each study year, when 10 to 33% of the samples were positive for campylobacter. The same PFGE genotypes found in humans were also detected in the chicken samples. This suggests that common genotypes were circulating in the area. PMID- 10790139 TI - Occurrence of Bacteroides fragilis enterotoxin gene-carrying strains in Germany and the United States. AB - Ninety-three Bacteroides fragilis isolates from different geographic locations were analyzed for the presence of an enterotoxin-encoding gene. It was shown that blood culture isolates were more likely to carry this gene than strains from other sources. All enterotoxin-positive strains belonged to the PCR fingerprint group I. PMID- 10790141 TI - Multiplex PCRs for identification of Escherichia coli virulence genes. AB - PCRs were developed to detect 11 Escherichia coli virulence genes. Primers amplified the respective genes without cross-reaction with other genes. Specificity was maintained in multiplex reactions; excellent amplification of target genes was possible with a minimum of four multiplex reactions. These reactions successfully identified genes in E. coli from the feces of four dogs. PMID- 10790142 TI - Brucella abortus infection acquired in microbiology laboratories. AB - We report an outbreak of laboratory-acquired Brucella abortus infection originating in the accidental breakage of a centrifuge tube. A total of 12 laboratory workers were infected (attack rate of 31%), with an incubation time ranging from 6 weeks to 5 months. Antibody titers were evaluated weekly in all personnel exposed, allowing the diagnosis of the infection in most cases before the onset of clinical symptoms, so that specific therapy could be administrated. PMID- 10790143 TI - Direct PCR of Cryptococcus neoformans MATalpha and MATa pheromones to determine mating type, ploidy, and variety: a tool for epidemiological and molecular pathogenesis studies. AB - Cryptococcus neoformans MATalpha and MATa pheromones were amplified by direct PCR. Nucleotide sequence analyses revealed unique restriction enzyme sites. Sixty strains were used to devise a restriction fragment length polymorphism typing scheme that yielded three variety-specific patterns. Additionally, pheromone specific PCR allowed easier identification of diploid C. neoformans strains than flow cytometry. PMID- 10790144 TI - Bilateral pulmonary aspergilloma caused by an atypical isolate of Aspergillus terreus. AB - A case of bilateral pulmonary aspergilloma caused by an atypical isolate of Aspergillus terreus is described. The diagnosis was established by the presence of septate, dichotomously branched fungal elements in freshly collected bronchoalveolar lavage and sputum specimens and by repeated isolation of the fungus in culture. Specific precipitating antibodies against the A. terreus isolate were demonstrated in the patient's serum. The isolate was atypical as it failed to produce fruiting structures on routine mycological media, but it did so on extended incubation on potato flake agar and produced globose, relatively heavy-walled, hyaline accessory conidia (formerly termed aleurioconidia) on both vegetative and aerial mycelia. Also, it produced an intense yellow diffusing pigment in the medium. The report underscores the increasing importance of A. terreus in the etiology of pulmonary aspergillosis. It is suggested that A. terreus antigen be included in the battery of serodiagnostic reagents to facilitate the early diagnosis of infections caused by this species. PMID- 10790145 TI - Infective endocarditis complicated with progressive heart failure due to beta lactamase-producing Cardiobacterium hominis. AB - We describe a 66-year-old woman with infective endocarditis due to Cardiobacterium hominis whose condition, complicated by severe aortic regurgitation and congestive heart failure, necessitated aortic valve replacement despite treatment with ceftriaxone followed by ciprofloxacin. The blood isolate of C. hominis produced beta-lactamase and exhibited high-level resistance to penicillin (MIC, >==256 microgram/ml) and reduced susceptibility to vancomycin (MIC, 8 microgram/ml). PMID- 10790146 TI - Isolation of an unusual Mycobacterium species from an AIDS patient with acute lymphadenitis. AB - A nonchromogenic Mycobacterium species was isolated from an AIDS patient with acute lymphadenitis. On the basis of the results of conventional tests, the strain appeared to be an atypical nonphotochromogenic Mycobacterium kansasii strain. Sequencing of the 16S rRNA gene revealed a unique nucleic acid sequence, suggesting that the isolate represents an undescribed pathogenic species. PMID- 10790148 TI - Deletion of phenylalanine 508 causes attenuated phosphorylation-dependent activation of CFTR chloride channels. AB - In cell-attached patches stimulated with cAMP agonists, the single-channel open probability (Po) of the phenylalanine 508-deleted cystic fibrosis transmembrane conductance regulator (DeltaF508-CFTR) channel, the most common disease associated mutation in cystic fibrosis, was abnormally low (a functional defect). To investigate the mechanism for the poor response of DeltaF508-CFTR to cAMP stimulation, we examined, in excised inside-out patches, protein kinase A (PKA) dependent phosphorylation activation and ATP-dependent gating of wild-type (WT) and DeltaF508-CFTR channels expressed in NIH3T3 mouse fibroblasts. For WT-CFTR, the activation time course of CFTR channel current upon addition of PKA and ATP followed a sigmoidal function with time constants that decreased as [PKA] was increased. The curvilinear relationship between [PKA] and the apparent activation rate suggests an incremental phosphorylation-dependent activation of CFTR at multiple phosphorylation sites. The time course of PKA-dependent activation of DeltaF508-CFTR channel current also followed a sigmoidal function, but the rate of activation was at least 7-fold slower than that with WT channels. This result suggests that deletion of phenylalanine 508 causes attenuated PKA-dependent phosphorylation of the CFTR chloride channel. Once DeltaF508-CFTR channels were maximally activated with PKA, the mutant channel and WT channel had indistinguishable steady-state Po values, ATP dose-response relationships and single-channel kinetics, indicating that DeltaF508-CFTR is not defective in ATP dependent gating. By measuring whole-cell current density, we compared the number of functional channels in WT- and DeltaF508-CFTR cell membrane. Our data showed that the estimated channel density for DeltaF508-CFTR was approximately 10-fold lower than that for WT-CFTR, but the cAMP-dependent whole-cell current density differed by approximately 200-fold. We thus conclude that the functional defect (a decrease in Po) of DeltaF508-CFTR is as important as the trafficking defect (a decrease in the number of functional channels in the plasma membrane) in cystic fibrosis pathogenesis. PMID- 10790147 TI - Regulation of volume-activated chloride channels by P-glycoprotein: phosphorylation has the final say! AB - P-glycoprotein (Pgp) is a transmembrane transporter causing efflux of a number of chemically unrelated drugs and is responsible for resistance to a variety of anticancer drugs during chemotherapy. Pgp overexpression in cells is also associated with volume-activated chloride channel activity; Pgp is thought to regulate such activity. Reversible phosphorylation is a possible mechanism for regulating the transport and chloride channel regulation functions of Pgp. Protein kinase C (PKC) is a good candidate for inducing such phosphorylation. Hierarchical multiple phosphorylation (e.g. of different serines and with different PKC isoforms) may shuttle the protein between its different states of activity (transport or channel regulation). Cell volume changes may trigger phosphorylation of Pgp at sites causing inhibition of transport. The possible regulation of chloride channels by Pgp and the potential involvement of reversible phosphorylation in such regulation is reviewed. PMID- 10790149 TI - Structural domains of the human GABAA receptor 3 subunit involved in the actions of pentobarbital. AB - This study was conducted to search for the residues of the beta3 subunit which affect pentobarbital action on the gamma-aminobutyric acid type A (GABAA) receptor. Three chimeras were constructed by joining the GABAA receptor beta3 subunit to the rho1 subunit. For each chimera, the N-terminal sequence was derived from the beta3 subunit and the C-terminal sequence from the rho1 subunit, with junctions located between the membrane-spanning regions M2 and M3, in the middle of M2, or in M1, respectively. In receptors obtained by the coexpression of alpha1 with the chimeric subunits, in contrast with those obtained by the coexpression of alpha1 and beta3, pentobarbital exhibited lower potentiation of GABA-evoked responses, and in the direct gating of Cl- currents, an increase in the EC50 together with a marked decrease in the relative maximal efficacy compared with that of GABA. Estimates of the channel opening probability through variance analysis and single-channel recordings of one chimeric subunit showed that the reduced relative efficacy for gating largely resulted from an increase in gating by GABA, with little change in efficacy of pentobarbital. A fit of the time course of the response by the predictions of a class of reaction schemes is consistent with the conclusion that the change in the concentration dependence of activation by pentobarbital is due to a change in pentobarbital affinity for the receptor. Therefore, the data suggest that residues of the beta3 subunit involved in pentobarbital binding to GABAA receptors are located downstream from the middle of the M2 region. PMID- 10790150 TI - GABAergic mIPSCs in rat cerebellar Purkinje cells are modulated by TrkB and mGluR1-mediated stimulation of Src. AB - Whilst protein tyrosine kinase (PTK) activity can modulate expressed GABAA receptors in cell culture, the physiological consequences on synaptic GABAA receptors are unknown. This was examined using whole-cell recording of bicuculline-sensitive mIPSCs in Purkinje cells (PCs) in cerebellar slices. Postsynaptic application of a peptide activator of the non-receptor PTK Src (Src peptide) enhanced mIPSC amplitudes by 39 % in the presence of brain-derived neurotrophic factor (BDNF) only; neurotrophin-3 (NT-3) was ineffective in this regard. Thus Src and TrkB (the receptor for BDNF) can physiologically interact to modulate synaptic GABAA receptors. In the presence of BDNF, pharmacological activation of metabotrophic glutamate receptor subtype 1 (mGluR1) by (S)-3, 5 dihydrophenylglycine (3,5-DHPG) also lead to a 32 % enhancement of mIPSCs. This enhancement was blocked by intracellular dialysis of PCs with PP1, a selective inhibitor of Src. It is concluded that, whilst GABAA receptors are not constitutively regulated by endogenous PTK activity in PCs, co-activation of TrkB by BDNF and Src by mGluR1 is required to modulate GABAergic synapses in PCs. PMID- 10790151 TI - The subunit dominates the relaxation kinetics of heteromeric neuronal nicotinic receptors. AB - The ACh-induced voltage-jump relaxation currents of the nicotinic receptors formed by pair-wise expression of the rat alpha2, alpha3, or alpha4 subunits with the beta2 or beta4 subunit in Xenopus oocytes were fitted best by the sum of two exponentials and a constant between -60 and -150 mV. As the ACh concentration approached zero, the relaxation time constants approached limiting values that should equal the single-channel burst duration at low ACh concentrations and the synaptic current decay time constants. beta4 co-expression prolonged the zero ACh concentration limits for the relaxation time constants. The fast beta4 zero ACh concentration limits ranged from 40 to 121 ms between -60 and -150 mV, and the slow beta4 zero ACh concentration limits ranged from 274 to 1039 ms. In contrast, the fast beta2 limits were 4-6 ms over the same voltage range and the slow beta2 limits were 30-53 ms. Expression with the beta4 subunit increased the voltage sensitivity of the alpha2, alpha3 and slow alpha4 relaxation time constants but not that of the fast alpha4 relaxation time constant. Reducing the temperature from 22 C to 8-9 C increased the alpha4beta2 and alpha3beta4 relaxation time constants 2.3- to 6.6-fold and reduced the fractional amplitude of the fast relaxation component. It also increased the voltage dependence of the fast alpha3beta4 relaxation time constant and decreased that of the slow time constant. The Q10 for alpha4beta2 and alpha3beta4 relaxation time constants ranged from 1.9 to 3.9 between 10 and 20 C. The beta subunit appears to have a dominant influence on the voltage-jump relaxation kinetics of heteromeric neuronal nicotinic receptors. PMID- 10790152 TI - Histamine-induced Ca2+ oscillations in a human endothelial cell line depend on transmembrane ion flux, ryanodine receptors and endoplasmic reticulum Ca2+ ATPase. AB - Using single cell microfluorometry to monitor changes in bulk Ca2+ concentration ([Ca2+]bulk) and the whole-cell configuration of the patch clamp technique to measure K+ currents (voltage clamp) and membrane potential (current clamp), the mechanisms of histamine-induced Ca2+ oscillations in the umbilical vein endothelial cell-derived cell line EA.hy926 were studied. In single cells, histamine (10 microM) evoked sinusoidal Ca2+ oscillations in low extracellular Ca2+ concentrations ([Ca2+]o = 10-30 microM). In contrast, histamine did not initiate Ca2+ oscillations either in the absence of extracellular Ca2+ (10 microM EGTA) or in the presence of 2.5 mM extracellular Ca2+. Ca2+ oscillations were accompanied by rhythmic activation of Ca2+-activated K+ (KCa) channels and membrane hyperpolarization of 18.1 +/- 3.9 mV. Hence, cell depolarization with 70 mM extracellular K+ or the inhibition of non-selective cation channels (NSCCs) and KCa channels by 10 microM Loe 908 and 10 mM tetrabutylammonium prevented histamine-evoked Ca2+ oscillations. Preventing Na+-Ca2+ exchange (NCX) by 10 microM 2', 4'-dichlorobenzamil, or removal of extracellular Na+, abolished histamine-induced Ca2+ oscillations. Lowering the extracellular Na+ concentration and thus promoting the reversed mode of NCX (3Na+ out and 1Ca2+ in) increased the amplitude and frequency of histamine-induced Ca2+ oscillations by 25 and 13 %, respectively. Hence, in the absence of extracellular Ca2+, 10 microM histamine induced an elevation of intracellular Na+ concentration in certain subplasmalemmal domains. The inhibitor of sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) 2,5-di-tert-butyl-1, 4-benzo-hydroquinone (15 microM) prevented histamine induced Ca2+ oscillations. In addition, blockage of ryanodine-sensitive Ca2+ release (RsCR) by 25 microM ryanodine blunted Ca2+ oscillations. In endothelial cells that were treated for 16 h with 10 microM nocodazole to collapse the superficial endoplasmic reticulum (sER), no histamine-induced Ca2+ oscillations were found. We conclude that in low [Ca2+]o conditions histamine-induced Ca2+ oscillations depend on transmembrane Na+ loading through NSCCs that leads to Ca2+ entry via NCX. Cation influx is controlled by KCa channel activity that triggers membrane hyperpolarization and, thus, provides the driving force for cation influx. Hence, the Ca2+ entering needs to be sequestrated via SERCA into sER to become released by RsCR to evoke Ca2+ spiking. These data further support our previous work on localized Ca2+ signalling as a key phenomenon in endothelial Ca2+ homeostasis. PMID- 10790153 TI - Subplasmalemmal ryanodine-sensitive Ca2+ release contributes to Ca2+-dependent K+ channel activation in a human umbilical vein endothelial cell line. AB - The whole-cell configuration of the patch clamp technique was used to assess the involvement of ryanodine-sensitive Ca2+ release (RsCR) in histamine-activated Ca2+-dependent K+ (KCa) channels in the human umbilical vein endothelial cell line EA.hy926. Histamine (10 microM) induced a transient outward current that reached 18.9 +/- 5.5 pA pF-1 at +20 mV. This current was diminished by 1 mM tetraethylammonium or 50 nM iberiotoxin, by 90 % and 80 %, respectively, suggesting that this current results from the stimulation of large-conductance KCa (BKCa) channels. In about 50 % of the cells tested, stimulation of RsCR with 200 nM ryanodine initiated a small outward current that was also sensitive to iberiotoxin. Following the ryanodine-mediated RsCR, the potency of 10 microM histamine to activate KCa channels was reduced by about 60 %. In agreement, an inhibition of RsCR with 25 microM ryanodine diminished KCacurrent in response to histamine by about 70 %. The effect of 100 microM histamine on KCa channel activity was not reduced by previous RsCR with 200 nM ryanodine, or by an inhibition of RsCR by 25 microM ryanodine. Histamine (10 microM)-induced Ca2+ elevation was reduced by 30 % following ryanodine-mediated RsCR, whereas no inhibition occurred in the case of 100 microM histamine stimulation. In cells treated with 10 microM nocodazole for 16 h to collapse the superficial endoplasmic reticulum, 200 nM ryanodine failed to initiate any KCa current. Furthermore, the inhibitory effect of previous RsCR on 10 microM histamine induced KCa current was not obtained in nocodazole-treated cells. Our data suggest that during moderate cell stimulation (10 microM histamine), subplasmalemmal RsCR greatly contributes to the activation of KCa channels in endothelial cells. Thus, the function of the subplasmalemmal Ca2+ control unit (SCCU) described previously must be extended as a regulator for KCa channels. PMID- 10790154 TI - Modulation of kir4.1 and kir5.1 by hypercapnia and intracellular acidosis. AB - CO2 chemoreception may be mediated by the modulation of certain ion channels in neurons. Kir4.1 and Kir5.1, two members of the inward rectifier K+ channel family, are expressed in several brain regions including the brainstem. To test the hypothesis that Kir4.1 and Kir5. 1 are modulated by CO2 and pH, we carried out experiments by expressing Kir4.1 and coexpressing Kir4.1 with Kir5.1 (Kir4.1 Kir5. 1) in Xenopus oocytes. K+ currents were then studied using two-electrode voltage clamp and excised patches. Exposure of the oocytes to CO2 (5, 10 and 15 %) produced a concentration-dependent inhibition of the whole-cell K+ currents. This inhibition was fast and reversible. Exposure to 15 % CO2 suppressed Kir4.1 currents by approximately 20 % and Kir4.1-Kir5.1 currents by approximately 60 %. The effect of CO2 was likely to be mediated by intracellular acidification, because selective intracellular, but not extracellular, acidification to the measured hypercapnic pH levels lowered the currents as effectively as hypercapnia. In excised inside-out patches, exposure of the cytosolic side of membranes to solutions with various pH levels brought about a dose-dependent inhibition of the macroscopic K+ currents. The pK value (-log of dissociation constant) for the inhibition was 6.03 in the Kir4.1 channels, while it was 7.45 in Kir4.1-Kir5.1 channels, an increase in pH sensitivity of 1.4 pH units. Hypercapnia without changing pH did not inhibit the Kir4.1 and Kir4.1-Kir5.1 currents, suggesting that these channels are inhibited by protons rather than molecular CO2. A lysine residue in the N terminus of Kir4.1 is critical. Mutation of this lysine at position 67 to methionine (K67M) completely eliminated the CO2 sensitivity of both the homomeric Kir4. 1 and heteromeric Kir4.1-Kir5.1. These results therefore indicate that the Kir4.1 channel is inhibited during hypercapnia by a decrease in intracellular pH, and the coexpression of Kir4.1 with Kir5.1 greatly enhances channel sensitivity to CO2/pH and may enable cells to detect both increases and decreases in PCO2 and intracellular pH at physiological levels. PMID- 10790155 TI - Slow modal gating of single G protein-activated K+ channels expressed in Xenopus oocytes. AB - The slow kinetics of G protein-activated K+ (GIRK) channels expressed in Xenopus oocytes were studied in single-channel, inside-out membrane patches. Channels formed by GIRK1 plus GIRK4 subunits, which are known to form the cardiac acetylcholine (ACh)-activated GIRK channel (KACh), were activated by a near saturating dose of G protein betagamma subunits (Gbetagamma; 20 nM). The kinetic parameters of the expressed GIRK1/4 channels were similar to those of cardiac KACh. GIRK1/4 channels differed significantly from channels formed by GIRK1 with the endogenous oocyte subunit GIRK5 (GIRK1/5) in some of their kinetic parameters and in a 3-fold lower open probability, Po. The unexpectedly low Po (0.025) of GIRK1/4 was due to the presence of closures of hundreds of milliseconds; the channel spent approximately 90 % of the time in the long closed states. GIRK1/4 channels displayed a clear modal behaviour: on a time scale of tens of seconds, the Gbetagamma-activated channels cycled between a low-Po mode (Po of about 0.0034) and a bursting mode characterized by an approximately 30-fold higher Po and a different set of kinetic constants (and, therefore, a different set of channel conformations). The available evidence indicates that the slow modal transitions are not driven by binding and unbinding of Gbetagamma. The GTPgammaS activated Galphai1 subunit, previously shown to inhibit GIRK channels, substantially increased the time spent in closed states and apparently shifted the channel to a mode similar, but not identical, to the low-Po mode. This is the first demonstration of slow modal transitions in GIRK channels. The detailed description of the slow gating kinetics of GIRK1/4 may help in future analysis of mechanisms of GIRK gating. PMID- 10790156 TI - Characterisation of a cell swelling-activated K+-selective conductance of ehrlich mouse ascites tumour cells. AB - The K+ and Cl- currents activated by hypotonic cell swelling were studied in Ehrlich ascites tumour cells using the whole-cell recording mode of the patch clamp technique. Currents were measured in the absence of added intracellular Ca2+ and with strong buffering of Ca2+. K+ current activated by cell swelling was measured as outward current at the Cl- equilibrium potential (ECl) under quasi physiological gradients. It could be abolished by replacing extracellular Na+ with K+, thereby cancelling the driving force. Replacement with other cations suggested a selectivity sequence of K+ > Rb+ > NH4 approximately Na+ approximately Li+; Cs+ appeared to be inhibitory. The current-voltage relationship of the volume-sensitive K+ current was well fitted with the Goldman Hodgkin-Katz current equation between -130 and +20 mV with a permeability coefficient of around 10(-6) cm s(-1) with both physiological and high-K+ extracellular solutions. The class III antiarrhythmic drug clofilium blocked the volume-sensitive K+ current in a voltage-independent manner with an IC50 of 32 microM. Clofilium was also found to be a strong inhibitor of the regulatory volume decrease response of Ehrlich cells. Cell swelling-activated K+ currents of Ehrlich cells are voltage and calcium insensitive and are resistant to a range of K+ channel inhibitors. These characteristics are similar to those of the so called background K+ channels. Noise analysis of whole-cell current was consistent with a unitary conductance of 5.5 pS for the single channels underlying the K+ current evoked by cell swelling, measured at 0 mV under a quasi physiological K+ gradient. PMID- 10790158 TI - O2-sensitive K+ channels: role of the Kv1.2 -subunit in mediating the hypoxic response. AB - One of the early events in O2 chemoreception is inhibition of O2-sensitive K+ (KO2) channels. Characterization of the molecular composition of the native KO2 channels in chemosensitive cells is important to understand the mechanism(s) that couple O2 to the KO2 channels. The rat phaeochromocytoma PC12 clonal cell line expresses an O2-sensitive voltage-dependent K+ channel similar to that recorded in other chemosensitive cells. Here we examine the possibility that the Kv1.2 alpha-subunit comprises the KO2 channel in PC12 cells. Whole-cell voltage-clamp experiments showed that the KO2 current in PC12 cells is inhibited by charybdotoxin, a blocker of Kv1.2 channels. PC12 cells express the Kv1.2 alpha subunit of K+ channels: Western blot analysis with affinity-purified anti-Kv1.2 antibody revealed a band at approximately 80 kDa. Specificity of this antibody was established in Western blot and immunohystochemical studies. Anti-Kv1.2 antibody selectively blocked Kv1.2 current expressed in the Xenopus oocyte, but had no effect on Kv2.1 current. Anti-Kv1.2 antibody dialysed through the patch pipette completely blocked the KO2 current, while the anti-Kv2.1 and irrelevant antibodies had no effect. The O2 sensitivity of recombinant Kv1.2 and Kv2.1 channels was studied in Xenopus oocytes. Hypoxia inhibited the Kv1.2 current only. These findings show that the KO2 channel in PC12 cells belongs to the Kv1 subfamily of K+ channels and that the Kv1.2 alpha-subunit is important in conferring O2 sensitivity to this channel. PMID- 10790157 TI - mu-Opioid receptor inhibits N-type Ca2+ channels in the calyx presynaptic terminal of the embryonic chick ciliary ganglion. AB - A study was made on the mechanisms by which enkephalins inhibit synaptic transmission at calyx-type presynaptic terminals in the ciliary ganglion of chick embryos at stages 39-40. Excitatory postsynaptic currents (EPSCs) were recorded by nystatin-perforated patch clamp at low [Ca2+]o and high [Mg2+]o. [Leu5]enkephalin (L-ENK, 1-10 microM) reduced the quantal content (m) without changing the quantal size (q). This effect was antagonized by naloxone (1 microM). Similar results were observed under conventional whole-cell clamp of the postsynaptic neuron. A specific agonist of the mu-opioid receptor, [D-Ala2, M-Me Phe4,Gly5]enkephalin-ol (DAMGO) reduced m without changing q. A specific agonist of the delta-opioid receptor, [d-Pen2, d-Pen5]enkephalin (DPDPE) also reduced m without changing q. Both L-ENK and [Met5]enkephalin (M-ENK) reduced the stimulus dependent increment of the intraterminal Ca2+ concentration (Delta[Ca2+]t) without affecting the decay time constant of the intraterminal Ca2+ concentration and basal Ca2+ level. This effect was antagonized by naloxone. DAMGO reduced Delta[Ca2+]t more effectively than DPDPE. When extracellular Ca2+ was replaced by Ba2+, the stimulus-dependent increment of the intraterminal Ba2+ concentration (Delta[Ba2+]t) was also reduced by L-ENK or DAMGO. L-ENK reduced Delta[Ca2+]t even in the presence of 4-aminopyridine (4-AP), which blocks the transient K+ conductance during the falling phase of the presynaptic action potential. When N type Ca2+ channels were blocked by omega-conotoxin GVIA (omega-CgTxGVIA), the Delta[Ca2+]t was no longer sensitive to L-ENK and DAMGO. It is suggested that enkephalins reduce the transmitter release through presynaptic opioid receptors. The mu-opioid receptor may suppress presynaptic Ca2+ influx by selectively inhibiting N-type Ca2+ channels. PMID- 10790159 TI - Functional coupling between glycolysis and excitation-contraction coupling underlies alternans in cat heart cells. AB - Electromechanical alternans was characterized in single cat atrial and ventricular myocytes by simultaneous measurements of action potentials, membrane current, cell shortening and changes in intracellular Ca2+ concentration ([Ca2+]i). Using laser scanning confocal fluorescence microscopy, alternans of electrically evoked [Ca2+]i transients revealed marked differences between atrial and ventricular myocytes. In ventricular myocytes, electrically evoked [Ca2+]i transients during alternans were spatially homogeneous. In atrial cells Ca2+ release started at subsarcolemmal peripheral regions and subsequently spread toward the centre of the myocyte. In contrast to ventricular myocytes, in atrial cells propagation of Ca2+ release from the sarcoplasmic reticulum (SR) during the small-amplitude [Ca2+]i transient was incomplete, leading to failures of excitation-contraction (EC) coupling in central regions of the cell. The mechanism underlying alternans was explored by evaluating the trigger signal for SR Ca2+ release (voltage-gated L-type Ca2+ current, ICa,L) and SR Ca2+ load during alternans. Voltage-clamp experiments revealed that peak ICa,L was not affected during alternans when measured simultaneously with changes of cell shortening. The SR Ca2+ content, evaluated by application of caffeine pulses, was identical following the small-amplitude and the large-amplitude [Ca2+]i transient. These results suggest that the primary mechanism responsible for cardiac alternans does not reside in the trigger signal for Ca2+ release and SR Ca2+ load. beta-Adrenergic stimulation with isoproterenol (isoprenaline) reversed electromechanical alternans, suggesting that under conditions of positive cardiac inotropy and enhanced efficiency of EC coupling alternans is less likely to occur. The occurrence of electromechanical alternans could be elicited by impairment of glycolysis. Inhibition of glycolytic flux by application of pyruvate, iodoacetate or beta-hydroxybutyrate induced electromechanical and [Ca2+]i transient alternans in both atrial and ventricular myocytes. The data support the conclusion that in cardiac myocytes alternans is the result of periodic alterations in the gain of EC coupling, i. e. the efficacy of a given trigger signal to release Ca2+ from the SR. It is suggested that the efficiency of EC coupling is locally controlled in the microenvironment of the SR Ca2+ release sites by mechanisms utilizing ATP, produced by glycolytic enzymes closely associated with the release channel. PMID- 10790160 TI - Endothelin-1 and photoreleased diacylglycerol increase L-type Ca2+ current by activation of protein kinase C in rat ventricular myocytes. AB - The amphotericin B-perforated whole-cell patch clamp technique was used to determine the modulation of L-type Ca2+ channels by protein kinase C (PKC) mediated pathways in adult rat ventricular myocytes. Application of 10 nM endothelin-1 (ET-1) increased peak Ca2+ current (ICa) by 28.2 +/- 2.5 % (n = 13) and slowed current decay. These effects were prevented by the endothelin receptor antagonist PD145065 (10 microM) and by the PKC inhibitor chelerythrine (8 microM). To establish if direct activation of PKC mimicked the ET-1 effect, the active and inactive phorbol esters (phorbol-12-myristate-13-acetate and 4alpha phorbol-12, 13-didecanoate) were tested. Both phorbol esters (100 nM) resulted in a small (approximately 10%) increase in ICa, suggesting PKC-independent effects. Bath application of dioctanoylglycerol (diC8), a diacylglycerol (DAG) analogue which is capable of directly activating PKC, caused a gradual decline in peak ICa (50.4 +/- 6.2 %, n = 5) and increased the rate of current decay. These effects were unaffected by the PKC inhibitor chelerythrine (8 microM). Intracellular photorelease of caged diC8 with 3 or 10 s exposure to UV light produced a concentration-dependent increase in peak ICa (20. 7 +/- 8.5 % (n = 8) for 3 s UV and 60.8 +/- 11.4 % (n = 13) for 10 s UV), which could be inhibited by chelerythrine. Our results demonstrate that both ET-1 and intracellularly photoreleased diC8 increase ICa by a PKC-mediated pathway, which is in direct contrast to the PKC-independent inhibition of ICa produced by bath-applied diC8. We conclude that specific cellular pools of DAG are crucially important in the regulation of ICa by PKC. PMID- 10790161 TI - Ca2+ signalling in rat vascular smooth muscle cells: a role for protein kinase C at physiological vasoconstrictor concentrations of vasopressin. AB - Physiological vasoconstrictor concentrations of Arg8-vasopressin (AVP, 10-100 pM) stimulate oscillations (spikes) in cytosolic free Ca2+ concentration ([Ca2+]i) in A7r5 rat vascular smooth muscle cells. These Ca2+ spikes are dependent on L-type voltage-sensitive Ca2+ channels and increase in frequency with increasing AVP concentration. The signal transduction pathway responsible for this effect was examined in fura-2-loaded A7r5 cell monolayers. The serine/threonine phosphatase inhibitor calyculin A (5 nM) sensitized A7r5 cells to AVP, resulting in the stimulation of Ca2+ spiking by 1-10 pM AVP. Calyculin A alone did not stimulate Ca2+ spiking. The protein kinase C (PKC) activator 4beta-phorbol 12-myristate 13 acetate (PMA, 100 pM to 200 nM), also stimulated Ca2+ spiking and this effect was additive with a submaximal concentration of AVP (50 pM). The PKC inhibitors Ro-31 8220 (1 microM) and calphostin C (250 nM) completely blocked the stimulation of Ca2+ spiking by either PMA or AVP. alpha, beta, gamma, delta, epsilon, zeta and &lamdda; isoforms of PKC were detected in A7r5 cells by Western blot analysis. Time-dependent redistribution of PKC-alpha, -delta and -epsilon isoforms between the membrane and cytosolic fractions occurred in response to 100 pM AVP. Pretreatment for 24 h with 1 microM PMA downregulated expression of PKC-alpha and -delta, but not PKC-epsilon, and prevented the Ca2+-spiking responses to either 1 nM PMA or 100 pM AVP. Neither the release of intracellular Ca2+ by 1 microM AVP nor the increase in [Ca2+]i in response to elevated extracellular [K+] was prevented by the PMA pretreatment. We conclude that PKC activation is a necessary step in the signal transduction pathway linking low concentrations of AVP to Ca2+ spiking in A7r5 cells. PMID- 10790162 TI - Serotonin suppresses subthreshold and suprathreshold oscillatory activity of rat inferior olivary neurones in vitro. AB - The effect of serotonin on membrane potential oscillations of inferior olivary neurones was studied in brainstem slices from 10- to 19-day-old rats. Serotonin at 50 and 5 microM induced a mean depolarization of 9.4 and 7.7 mV, respectively, that was preceded by a reversible suppression of subthreshold membrane potential oscillations. These effects were not changed by 1 microM tetrodotoxin and the suppression of subthreshold oscillations persisted after current-mediated restoration of resting potential. In spontaneously active neurones, serotonin abolished the rhythmicity of action potential firing without affecting spike frequency. Serotonin reduced the slope of the calcium-mediated rebound spike and both the duration and amplitude of the subsequent afterhyperpolarization. Serotonin also shifted the voltage dependence of the rebound spike to more negative values. Hyperpolarizing current pulses (200 ms) revealed that serotonin increased the pre-rectification and steady-state components of membrane resistance by 37 and 38 %, respectively, in 66 % of neurones, but decreased these parameters by 14 and 20% in the remaining cells. The serotonin effects were antagonized by 5 microM methysergide or 1-5 microM ketanserin and were mimicked by 10-20 microM dimethoxy-4-iodoamphetamine but not 10 microM 8-hydroxy-2-(di-N propylamino)-tetralin. The data indicate that serotonin suppresses the rhythmic activity of olivary neurones via 5-HT2 receptors by inhibition of the T-type calcium current in combination with membrane depolarization due to activation of a cation current (Ih) and block of a resting K+ current (fast IK(ir)). This modulatory action of serotonin may account for the differential propensity of olivary neurones to fire rhythmically during different behavioural states in vivo. PMID- 10790163 TI - Evidence for myosin light chain kinase mediating noradrenaline-evoked cation current in rabbit portal vein myocytes. AB - The role of myosin light chain kinase (MLCK) in the activation of the noradrenaline-evoked non-selective cation current (Icat) was examined with the whole-cell recording technique in single rabbit portal vein smooth muscle cells. Intracellular dialysis with 5 microM MLCK(11-19)amide, a substrate-specific peptide inhibitor of MLCK, markedly reduced the amplitude and rate of activation of noradrenaline-evoked Icat. A similar result was obtained when the cells were dialysed with 10 microM AV25, which also inhibits MLCK by an action at the auto inhibitory domain of MLCK. Inhibitors of binding of ATP to MLCK, wortmannin and synthetic naphthalenesulphonyl derivatives (ML-7 and ML-9), at micromolar concentrations, also reduced the amplitude of noradrenaline-evoked Icat. ML-7 and ML-9 (both at 5 microM) reduced the amplitude of Icat induced by both guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) and 1-oleoyl-2-acetyl-sn-glycerol (OAG). MLCK(11-19)amide, AV25 and ML-9 did not inhibit the noradrenaline-evoked Ca2+ activated potassium current at a holding potential of 0 mV. In addition, MLCK(11 19)amide and AV25 did not reduce the non-selective cation current induced by ATP in rabbit ear artery cells. Intracellular dialysis with 2 microM Ca2+ and 9 microM calmodulin activated Icat, which developed over a period of about 5 min. Intracellular dialysis with the non-hydrolysable analogue of ATP, 5' adenylylimidodiphosphate (AMP-PNP), reduced the amplitude and rate of activation of noradrenaline-evoked Icat. The results indicate that MLCK mediates noradrenaline-activated Icat in rabbit portal vein smooth muscle cells. PMID- 10790164 TI - cGMP-mediated phosphorylation of heat shock protein 20 may cause smooth muscle relaxation without myosin light chain dephosphorylation in swine carotid artery. AB - Nitrovasodilators such as nitroglycerine, via production of nitric oxide and an increase in [cGMP], can induce arterial smooth muscle relaxation without proportional reduction in myosin light chain (MLC) phosphorylation or myoplasmic [Ca2+]. These findings suggest that regulatory systems, other than MLC phosphorylation and Ca2+, partially mediate nitroglycerine-induced relaxation. In swine carotid artery, we found that a membrane-permeant cGMP analogue induced relaxation without MLC dephosphorylation, suggesting that cGMP mediated the relaxation. Nitroglycerine-induced relaxation was associated with a reduction in O2 consumption, suggesting that the interaction between phosphorylated myosin and the thin filament was inhibited. Nitroglycerine-induced relaxation was associated with a 10-fold increase in the phosphorylation of a protein on Ser16. We identified this protein as heat shock protein 20 (HSP20), a member of a family of proteins known to bind to thin filaments. When homogenates of nitroglycerine relaxed tissues were centrifuged at 6000 g, phosphorylated HSP20 preferentially sedimented in the pellet, suggesting that phosphorylation of HSP20 may increase its affinity for the thin filament. We noted that a domain of HSP20 is partially homologous to the 'minimum inhibitory sequence' of skeletal troponin I. The peptide HSP20110-121, which contains this domain, bound to actin-containing filaments only in the presence of tropomyosin, a characteristic of troponin I. High concentrations of HSP20110-121 abolished Ca2+-activated force in skinned swine carotid artery. HSP20110-121 also partially decreased actin-activated myosin S1 ATPase activity. These data suggest that cGMP-mediated phosphorylation of HSP20 on Ser16 may have a role in smooth muscle relaxation without MLC dephosphorylation. HSP20 contains an actin-binding sequence at amino acid residues 110-121 that inhibited force production in skinned carotid artery. We hypothesize that phosphorylation of HSP20 regulates force independent of MLC phosphorylation via binding of HSP20 to thin filaments and inhibition of cross bridge cycling. PMID- 10790165 TI - Response characteristics and receptive field widths of on-bipolar cells in the mouse retina. AB - Voltage-clamp and current-clamp recordings were made from bipolar cells in dark adapted mouse retinal slices. Light-evoked responses fell into three groups corresponding to the rod bipolar cells, on-cone bipolar cells and off-cone bipolar cells. The morphology of the recorded cells confirmed this classification. Intensity-response relations were well fitted by a Michaelis saturation function with Hill coefficients of 1.15 +/- 0.11 (n = 6) for rod bipolar cells and 2.33 +/- 0.06 (n = 4) for cone inputs onto on-cone bipolar cells. In the absence of antagonists for GABA or glycine receptors, light-evoked synaptic currents for all cells displayed linear current-voltage relations that reversed near 0 mV, indicating that very little inhibition was activated under dark-adapted recording conditions. Saturating light stimuli evoked conductances of 0.81 +/- 0.56 nS (n = 4) in rod bipolar cells and 1.1 +/- 0.8 nS (n = 4) in on cone bipolar cells. Receptive field widths were estimated by flashing a vertical light bar at various locations along the slice. Rod and on-cone bipolar cells had receptive field widths of 67 +/- 16 micrometer (n = 6) and 43 +/- 7 microm (n = 5), respectively. The maximum spatial resolution of an array of such cone bipolar cells was estimated to be 0.3 cycles deg-1, compared with a maximum resolution of 0.5 cycles deg-1 obtained from behavioural studies in mice. Our results suggest that this limit to spatial resolution could be imposed early in the visual system by the size of the bipolar cell receptive fields. PMID- 10790166 TI - P2X purinoceptor-mediated excitation of trigeminal lingual nerve terminals in an in vitro intra-arterially perfused rat tongue preparation. AB - A novel in vitro intra-arterially perfused adult rat tongue-nerve preparation was used to explore the possible actions of P2X purinoceptor agonists (ATP and alpha,beta-methylene ATP (alpha, beta-meATP)) on sensory nerve terminals innervating the rat tongue. We made whole-nerve recordings of the trigeminal branch of the lingual nerve (LN), which conducts general sensory information (pain, temperature, touch, etc.), and the chorda tympani (CT), which conducts taste information. Changes in LN and CT activity following intra-arterial application of P2X agonists were compared. In seven preparations, bolus close arterial injection of ATP (30-3000 microM, 0.1 ml) or alpha,beta-meATP (10-300 microM, 0.1 ml) induced a rapid (< 1 s after injection), dose-related increase in LN activity that decayed within a few seconds. The minimal concentration of ATP (100 microM) required to elicit a response was about 10-fold higher than that of alpha,beta-meATP (10 microM). Bolus injection of ATP or alpha,beta-meATP induced a moderate decrease in firing frequency in three of seven CT preparations. LN responses to P2X agonists showed signs of rapid desensitisation with the peak frequency of discharge being smaller when the agonists were applied at short intervals. Suramin (200 microM) or PPADS (200 microM) applied by intra-arterial perfusion each antagonised the rapid increase in LN activity following application of alpha,beta-meATP (100 microM). Capsaicin (10 microM, 0.1 ml, n = 5 preparations) was injected intra-arterially to desensitise nociceptive fibres. This was found to block (n = 2) or greatly reduce (n = 3) the excitatory effects of alpha,beta-meATP (100 microM, 0.1 ml) on LN activity, implying that only capsaicin-sensitive nociceptive fibres in LN were responsive to P2X agonists. In contrast to the consistent excitatory responses in LN activity following fast application of P2X agonists as bolus, a variable and moderate change in discharge rate of LN and no change in CT activity (n = 5) was observed after applying ATP (100-300 microM, n = 21) or alpha,beta-meATP (100-300 microM, n = 14) by intra arterial perfusion. The variable responses in LN activity to slow perfusion in contrast to close-arterial bolus injection are consistent with activation of the rapidly desensitising P2X3 receptors. In summary, ATP and alpha,beta-meATP preferentially activate general sensory afferent fibres (LN) but not taste fibres (CT). We suggest that the increase in whole-nerve activity of LN following application of P2X agonists represents activation of nociceptive fibres which possess P2X3 receptors. Our data indicate that ATP and P2X3 receptors may play a role in nociception, rather than taste sensation in the tongue. PMID- 10790167 TI - Cardiovascular responses to carotid chemoreceptor stimulation in the dog: their modulation by urinary bladder distension. AB - Respiratory, heart rate and hindlimb vascular responses were studied in response to increasing levels of stimulation of the carotid body chemoreceptors, together with an examination of the modulation of their effects by distension of the urinary bladder in the dog anaesthetized with a mixture of chloralose and urethane. The vascularly isolated carotid bifurcation regions were perfused with blood, stimulation of the carotid bodies being carried out by three different levels of hypoxic isocapnic blood (PO2 approximately 58, 40 and 22 mmHg) obtained from a donor animal. A vascularly isolated hindlimb was autoperfused at constant blood flow through its femoral artery. In spontaneously breathing animals, increasingly intense hypoxic stimulation of the carotid bodies caused a progressive augmentation of respiratory minute volume. Superimposition of distension of the bladder increased ventilation further, by the same amount during hypoxic as during normoxic blood perfusion of the chemoreceptors. Prevention of the effects of lung stretch afferent stimulation by artificial ventilation modified the heart rate and hindlimb vascular responses to excitation of the carotid bodies by revealing or accentuating the primary cardiovascular responses, bradycardia and vasoconstriction. In contrast, no such respiratory modulation was apparent in the cardiovascular responses to bladder distension. When, under conditions of artificial ventilation and in the absence of changes in the arterial baroreceptor input, the primary cardio-inhibitory and vasoconstrictor responses to carotid chemoreceptor stimulation predominated, the heart slowed progressively as the stimulus was increased. At the same time the cardio-accelerator effects of bladder distension progressively diminished, indicating an interaction between the cardiac reflex responses evoked by the two inputs. In contrast, the reflex vascular responses resulting from stimulation of the two inputs were additive, at least for PO2 levels of carotid body perfusate down to approximately 40 mmHg. In conclusion these experiments demonstrate the differential nature of the integration of respiratory and cardiovascular responses evoked by stimulation of the carotid chemoreceptors and bladder distension. PMID- 10790168 TI - Post-exercise adipose tissue and skeletal muscle lipid metabolism in humans: the effects of exercise intensity. AB - One purpose of the present experiments was to examine whether the relative workload or the absolute work performed is the major determinant of the lipid mobilization from adipose tissue during exercise. A second purpose was to determine the co-ordination of skeletal muscle and adipose tissue lipid metabolism during a 3 h post-exercise period. Six subjects were studied twice. In one experiment, they exercised for 90 min at 40% of maximal O2 consumption (VO2,max) and in the other experiment they exercised at 60% VO2,max for 60 min. For both experiments, catheters were inserted in an artery, a subcutaneous abdominal vein and a femoral vein. Adipose tissue metabolism and skeletal muscle (leg) metabolism were measured using Fick's principle. The results show that the lipolytic rate in adipose tissue during exercise was the same in each experiment. Post-exercise, there was a very fast decrease in lipolysis, but it began to increase about 1 h post-exercise and remained elevated for the following 2 h. The increase in post-exercise non-esterified fatty acid (NEFA) mobilization was greater after 60% exercise than after 40 % exercise. It is concluded that the lipolytic rate in abdominal subcutaneous adipose tissue during exercise is the same whether the relative workload is 40% or 60% of maximum. Post-exercise, there is a substantial lipid mobilization from adipose tissue and only a small fraction of this is taken up in the lower extremities. This leaves a substantial amount of NEFAs for either NEFA/TAG (triacylglycerol) recirculation post-exercise or immediate oxidation. PMID- 10790170 TI - Prevention and treatment of painful neuromas of the superficial radial nerve by the end-to-side nerve repair concept: an experimental study and preliminary clinical experience. AB - This article studies the utilization of the end-to-side neurorrhaphy concept in the prevention and treatment of painful neuromas. A total of 20 rats were divided into 2 groups (10 rats per group). In group A, the tibial nerve was divided and left lying in the subcutaneous tissue. In group B, the cut ends of the tibial nerve were sutured to the adjacent peroneal nerve in an end-to-side fashion. Evaluation was performed 90 days after nerve injury. For group A, the proximal end of the tibial nerve formed a "classic" neuroma and the distal end showed a degenerated nerve. In group B, the proximal end of the tibial nerve formed a "non classic" neuroma and the nerve healed into the peroneal nerve with continuity of the epineurium of the 2 nerves. The distal end of the tibial nerve in group B showed evidence of axonal regeneration. Preliminary clinical experience utilizing the same technique in the prevention and treatment of painful neuromas of the superficial radial nerve is presented and other techniques of nerve-to-nerve implantation are discussed. PMID- 10790169 TI - Attenuation of vasodilatation with skeletal muscle fatigue in hamster retractor. AB - We tested the hypothesis that muscle fatigue would attenuate vasodilatory responsiveness throughout the resistance network. The retractor muscle of anaesthetized hamsters was contracted (once per 2 s for 1 min) at duty cycles of 2.5, 10 and 20 % before and after fatiguing contractions that diminished peak tension and muscle glycogen by >50 %. Arterioles and feed arteries (FA) dilated maximally during fatiguing contractions. Resting vasomotor tone consistently recovered following contractions. Peak blood flow was proportional to integrated tension (tension x time, expressed in mN mm-2 s); both increased with duty cycle and decreased with fatigue. Total integrated vasodilatory responses (diameter x time, expressed in microm s) increased with duty cycle and decreased with fatigue. Vasodilatation during contractions plateaued at approximately 50 % of peak integrated tension. Post-contraction vasodilatation increased with integrated tension and both were attenuated with fatigue. As integrated tension increased, distal arterioles dilated first and to the greatest extent relative to proximal arterioles and FA. Fatigue had little effect on dilatation of distal arterioles whereas dilatation of proximal arterioles and FA was suppressed. Latency of onset for vasodilatation decreased as duty cycle increased and was unaffected by fatigue. Vasodilatation and blood flow increase in proportion to integrated tension, with an ascending locus of vasomotor control and prolongation of post-contraction vasodilatation. With muscle fatigue, the locus of flow control resides in distal arterioles; both ascending and post-contraction vasodilatations are attenuated despite normal vasomotor tone. PMID- 10790171 TI - Tongue replantation in an animal model. AB - Although the tongue is not a vital organ in sustaining life, it may be a vital organ in sustaining the will to live in many people. As carcinoma of the tongue represents the majority of the 30,000 oral cavity cancers diagnosed per year in the United States, many patients face the potential consequences of resection of part or all of the tongue for cure. To date, reconstructive options do not restore optimal tongue function including articulation, swallowing, taste, or sensation. With the ultimate goal of improving tongue reconstruction, we report on a successfully performed autograft transplantation of the tongue in an animal model. Before undertaking allograft transplantation of the tongue, an autograft tongue transplant would be attempted to identify the feasibility of such a procedure and to determine the similarity of an animal model with human techniques. The dog's neck, tongue, and oral anatomy represent an excellent animal model for tongue reconstruction. This procedure can be performed successfully in an animal model. The only previously published replantation of the tongue involved the reattachment of the anterior portion of a human tongue after physical trauma. To our knowledge, the enclosed report represents the first successful total excision and replantation of the tongue in either a human or animal model. PMID- 10790172 TI - Rat multiple organ blocks: microsurgical technique of removal for ex vivo aerobic organ preservation using a fluorocarbon emulsion. AB - A multiple organ block (MOB) is composed of en bloc removed organs (heart, lungs, liver, pancreas, kidneys, and bowel), connected by the vascular system, of which blood circulation is maintained by the heart and oxygenation by the lungs under artificial ventilation. The aim of this study is the description of a surgical technique of MOB removal in the rat. Ninety-five MOBs were removed from Wistar rats. The rats were anesthetized, a tracheotomy was performed, and the cannula was connected to a pressure-regulated respirator. A colectomy was performed. Ureters, vena cava inferior, aorta, and bile duct were cannulated using an operative microscope. The vessels that joined the MOB to the carcass were tied or coagulated to make removal of the MOBs possible. Once removed, the MOBs were placed in a vaseline oil bath at 37 degrees C and the aorta and vena cava were connected to an accessory vascular circuit to stabilize arterious pressure. Success rate (ex vivo survival of more than 10 min) after the 30th attempt was 90% and after the 60th attempt was 95% (global success rate 82%). Ex vivo survival of MOBs at 37 degrees C ranged from 1 to 450 min. Rat MOBs allows us to study the normothermic preservation of all the organs susceptible of being transplanted in one single series of experiments. We showed that removal of rat MOBs is feasible. This microsurgical technique is codified. Rat MOBs are suitable if perfusion liquids are difficult to obtain or if a great number of experiments are required. As MOBs are composed of synergically functioning organs in the absence of striated muscle, bone, and nervous system, they also could be useful for physiologic and pharmacologic studies. PMID- 10790173 TI - Establishment of a parabiotic rat model by anastomosis of common carotid artery. AB - A cross circulation model was established by replacing mutual blood between two rats by anastomoses of each common carotid artery. In this model, parabiotic rats were shown to replace their mutual blood completely without any artificial materials or instruments, such as silicon tubes or pumps. The rate of blood exchange was measured by intravenous injection of Evans blue dye without the use of the radioactive materials that were so far commonly used. The partial alteration of blood circulation through microvascular surgery may reveal the mechanisms of organ tropism of bacteria, protozoa, and parasites. Microvascular surgery is available for reconstruction and analysis of organ functions as well as for clinical use in organplasty. PMID- 10790175 TI - Repair of nerve gap with the elongation of Wallerian degenerated nerve by tissue expansion. AB - We have found previously that expansion of the Wallerian degenerated nerve was accompanied by accelerated Schwann cell proliferation. In this study, we investigated the usefulness of the elongation of Wallerian degenerated nerve for the repair of short nerve gap. Male Wistar rats were used. After the left sciatic nerve was transected the rats were divided into 4 groups. In the control group, nerve coaptation was not performed. In group 1, tensionless coaptation was performed immediately. In group 2, delayed tensionless coaptation was performed with the elongation of Wallerian degenerated nerve. In group 3, coaptation was performed immediately with autologous interposition nerve graft. The ideal tensionless nerve repair of group 1 was considered to produce the best result. Rats in group 2 showed functional recovery as good as rats in group 1. On histologic assessment, in group 2, a fibrous capsule that was very rich in vascularity was formed around the tissue expander. After 14 weeks, the capsule was diminished markedly in size, but the vascularity was rich around the sciatic nerve. We think that the excellent functional recovery seen in group 2 can be attributed to the increased activity of Schwann cell proliferation and increased vascularity. PMID- 10790174 TI - Endothelium during microarterial graft procurement and transplantation. AB - Microarterial grafts are prone to mechanical endothelial injury that can have profound effects on the transplanted vessel, a factor neglected in studies on vascular changes post-transplantation. The aim of this study was to document the integrity of the endothelial lining after both procurement and transplantation of rat aortic grafts, using a minimal touch technique with and without the use of topical papaverine. It was found that procurement using a simple minimal touch technique preserved the endothelium and transplantation could be performed without endothelial injury. The appearance of "normal" endothelium varied with the degree of distension of the artery, suggesting a dynamic endothelial architecture to accommodate changes in the surface area of the artery during pulsation. These findings indicate that transplantation of an arterial segment without injury to the intima is possible and stress the importance of technical controls after both procurement and transplantation to prevent the use of injured grafts and misleading results. PMID- 10790176 TI - Nonpenetrating vascular clips for small-caliber anastomosis. AB - In the search for better anastomosing techniques, an improved vascular stapler device (VCS clip applier system(R)) has been introduced. The system uses nonpenetrating clips to approximate everted vessel walls. The objective of this study was to determine the effects of nonpenetrating vascular clips on endothelial wound healing. Aortic end-to-end anastomoses were performed in male Wistar rats. A comparison was made between clipped (n = 12) and conventional hand sewn (n = 6) anastomoses. Patency rates were verified at different time intervals (after 1, 4, and 8 weeks), after which the anastomotic sites were removed. Morphological evaluation was carried out using scanning electron microscopy. All rats survived the procedure. Closure with clips took less time than closure with conventional sutures, with decreasing aortic clamping times for the clipped procedures during the course of the experiments. Patency rates were 100% in both the "clipped" and "sutured" groups. Microscopic examination showed favorable endothelial healing at the clipped anastomotic sites, with less inflammatory reaction at 1 week, and a more complete endothelial regeneration at 4 and 8 weeks follow-up, as compared with the sutured anastomoses. The clip applier holds the promise of a useful device in anastomosing small-caliber vessels, since clip closure takes less time than suturing, while patency rates are identical, and morphological results are favorable. Training is mandatory to obtain technical skills and to achieve optimal results. PMID- 10790177 TI - Improved technique for infrahepatic vena cava reconstruction in guinea pig-to-rat liver transplantation. AB - Guinea pig-to-rat orthotopic liver transplantation is associated with serious technical problems contributing to impaired graft perfusion and primary graft failure. In order to shorten the procurement procedure and thereby minimize liver damage before flushing, a simplified technique for infrahepatic caval reconstruction was developed. Dissection of the infrahepatic vena cava (IHVC) from adrenal glands and renal and lumbar veins represents the most difficult and time-consuming part of the donor operation, which is often not well tolerated by the animal; we avoided this step by using an isogeneic vena cava interposition graft (VCIG) following in situ perfusion. This graft is connected with the IHVC transsected just below the liver with a cuff technique. Donor operations lasted 15 to 20 minutes with the new technique (n = 7) compared to 52 to 76 minutes with conventional technique (n = 7). Reduced operating time was associated with markedly improved graft perfusion and significantly better graft survival. This modification of the donor procedure for the guinea pig-to-rat liver xenograft using a VCIG significantly reduces operating time and improves reperfusion and recipient survival. PMID- 10790178 TI - Osteomyocutaneous flap as a preclinical composite tissue allograft: swine model. AB - Composite tissue allotransplantation (CTA) constitutes one of the last frontiers of microsurgery. Prior to its clinical application, the long-term efficacy of modern immunotherapy must be tested in a pre-clinical CTA model. Based on the concept of osteomyocutaneous forearm flap, we developed a CTA flap model in swine. After identifying the vascular territory of the flaps in six pigs (vascular casting), flaps were transplanted from mismatched donor to recipient pigs (n = 6). Rejection was assessed daily by visual inspection and histopathology of biopsy specimens. Recipient pigs were able to ambulate immediately following surgery. There were no flap failures owing to technical or surgical complications. Rejection occurred over a period of 7 days as manifested by edema, cellular infiltration, epidermalysis, and thrombosis. This pre-clinical flap model is excellent for evaluating the effectiveness of modern immunotherapy because it is anatomically and immunologically relevant and because the minimal morbidity caused to the animal permits long-term studies. PMID- 10790179 TI - Effect of CO(2)-Milliwatt laser on peripheral nerves: part II. A histological and functional study. AB - In order to further explore the role of laser for microneural repair, the early and late effects of CO(2) laser irradiation on intact rat sciatic nerves were investigated. A total of 48 rat sciatic nerves were exposed to 100-mW laser power with a pulse duration of 1.0 s and a spot size of 320 microm. In one-half of the nerves, albumin solder was applied to the nerve followed by laser irradiation. The results were evaluated up to 94 days after surgery with functional toe spreading test, and light and transmission electron microscopy. Irradiation of the nerve resulted in almost no deficit in the motor function. A subperineurial degeneration of myelinated and unmyelinated axons is observed in the first 2 weeks after laser irradiation, while the central part of the nerve remains undamaged. The degeneration is followed by axonal regeneration with subsequent maturation of nerve fibres in time. No excessive intraneural or extraneural scarring was seen. In the soldered nerves, the solder elicits an inflammatory reaction upon the epineurium in the first week after irradiation. By week 1, the solder is completely absorbed. After 2 weeks, the inflammatory reaction ceases and by week 4, no residual reaction is seen. At 12 weeks, only minimally disarranged epineurium is seen with otherwise normal neural architecture. In conclusion, CO(2) laser irradiation at 100 mW with pulses of 1.0 s has no long term negative effects on nerve function and morphology. Therefore, these laser settings can be safely applied for laser-assisted nerve repair. PMID- 10790180 TI - Neural inhibition of the corpus allatum in the last larval instar differs from that in adults in the blood-feeding insect, Rhodnius prolixus. AB - An active corpus allatum (CA) in the blood feeding insect, Rhodnius prolixus, releases a substance that inhibits metamorphosis in larvae, and activates egg production in adults. In adults, transecting the nervus corporis cardiacum II's (NCCII), which are attached posteriorly to each protocerebral lobe, greatly increases egg production indicating that the adult CA is activated and receives neural inhibition from cells associated with the NCCII [Chiang, Arch. Insect. Biochem. Physiol. 39:126-131 (1998)]. In the present study, the NCCII's in fifth instar larvae were transected immediately before or after feeding to determine if these nerves normally inhibit CA activity in the last larval instar. Approximately 20 to 25 days following ingestion of a blood meal, L5's with transected NCCII's emerged as fully-formed adults with no larval characteristics. Examination of the brain in these recently emerged adults revealed that the NCCII's were absent. Since fifth instar larvae with transected NCCII's emerged with no juvenile characteristics, cutting the NCCII's did not activate the CA, indicating that the mechanism for inhibition of the CA differs in the last larval instar and adult animals. PMID- 10790181 TI - Rhythmic autocrine activity in cultured insect epidermal cells. AB - It is now well established that ecdysteroids can be produced in insects in the absence of prothoracic glands. In this respect, it has been shown that cells in culture can produce ecdysteroids. Our aims were: (1) to determine whether ecdysteroid target cells of epidermal origin could also be the source of ecdysteroids; (2) to monitor more accurately the kinetics of ecdysteroid production; and (3) to check for possible relationships between this synthetic activity and dynamics of cell division. An insect cell line (IAL-PID2) established from imaginal discs of the Indian meal moth, Plodia interpunctella, with wild-type sensitivity to ecdysteroids was used in our study. Our results showed that the Plodia cell line exhibited autocrine activity. When division of IAL-PID2 cells was synchronized, a rhythmic production of ecdysteroids was observed. However, further experiments indicated that this rhythmicity could be cell autonomous. This led us to anticipate the existence of two cell subpopulations that would be able to produce ecdysteroids rhythmically, a minor one that would be cell cycle serum-independent population, and a major population that would need serum growth factors to proliferate and produce ecdysteroids. Qualitative study of the ecdysteroid content of the media clearly showed that ecdysone was the major immunoreactive product. Taken together, our findings clearly show that an insect cell line of epidermal origin is capable of rhythmic autocrine production of ecdysteroids. These results support the hypothesis that alternate sites for ecdysteroid production in vivo may exist and could play a role in local regulation of development. We now plan to determine the cellular basis of this rhythmic autocrine activity and to confirm the existence of growth factor-autonomous cells in the culture as well as the potent role played by ecdysteroids in the cross-talk between various cell subpopulations. PMID- 10790182 TI - A juvenile hormone-repressible transferrin-like protein from the bean bug, Riptortus clavatus: cDNA sequence analysis and protein identification during diapause and vitellogenesis. AB - We found several juvenile hormone-responsive cDNAs in the bean bug, Riptortus clavatus, by using mRNA differential display (Hirai et al., 1998). One of them, a juvenile hormone-repressible cDNA, JR-3, was cloned, sequenced, characterized and identified as a transferrin (RcTf). RcTf cDNA encoded 652 amino acids with a calculated molecular weight of 71,453 Da. The deduced amino acid sequence showed significant homology with the transferin genes of several insects, Manduca sexta (43% identity), Blaberus discoidalis (43%), Aedes aegypti (43%), Drosophila melanogaster (36%), Sarcophaga peregrina (36%) and the human (25%). Antiserum was prepared by using recombinant RcTf protein expressed in Escherichia coli as an antigen. The antiserum reacted specifically with both the recombinant protein and the native protein from the bugs, with sizes of 70 and 75 kDa, respectively. The 75 kDa protein was partially purified from hemolymph of diapausing female bugs and the first ten amino acids were found to be identical to that of RcTf cDNA, indicating that the 75 kDa protein is RcTf. The tissue distribution of RcTf in the bug was examined by Western blot analysis. In diapausing animals, RcTf was detected in the fat body, hemolymph and ovary but not in the gut. In the post diapause stage, RcTf was also detected in eggs, in addition to the fat body and ovary. These results indicate that RcTf is incorporated into the oocytes during vitellogenesis, and suggest that it may provide iron for the developing embryos. PMID- 10790183 TI - Calcium homeostasis in larval and adult Drosophila melanogaster. AB - Calcium homeostasis in Drosophila melanogaster was examined in response to the challenges imposed by growth, reproduction and variations in dietary calcium content. Turnover time for calcium, calculated as the time for (45)Ca(2+)to accumulate to half the steady state value of 3.46 nmol/fly, was 3.3 days. Although larvae weighed 2x as much as adults, they contained 3-4x as much calcium. Anterior Malpighian tubules (Mts) contain much more calcium than posterior Mts, accounting for 25-30% of the calcium content of the whole fly. In response to a 6.2-fold increase in dietary calcium level, calcium content of whole flies increased only 10%. Hemolymph calcium concentration ( approximately 0.5 mM) was similar in males and females and in animals raised on diets differing in calcium content. Fluid secretion rate, secreted fluid calcium concentration, and transepithelial calcium flux in tubules isolated from flies raised on high and low calcium diets did not differ significantly. Malpighian tubules secrete calcium at rates sufficient to eliminate whole body calcium content in 0.5 and 3 days for tubules secreting fluid at basal and maximal rates, respectively. It is suggested that flies absorb high quantities of calcium from the diet and maintain homeostasis through the combined effects of elimination of calcium in fluid secreted by the Malpighian tubules and the sequestration of calcium in granules, especially within the distal segment of the anterior pair of Malpighian tubules. PMID- 10790184 TI - DNA-binding proteins of the malaria vector Anopheles stephensi: purification and characterization of an endonuclease. AB - DNA-binding proteins present in fourth instar larvae of Anopheles stephensi were isolated by affinity chromatography on native and denatured DNA cellulose columns and analyzed by electrophoresis on polyacrylamide gels. A denatured DNA-specific protein with an approximate molecular weight of 30 kDa was the predominant DNA binding protein of larvae. This protein was purified to electrophoretic homogeneity by ammonium sulfate fractionation followed by phosphocellulose chromatography. The purified 30 kDa binding protein showed an endonucleolytic activity capable of converting pBR 322 supercoiled DNA to the circular form. Maximum endonucleolytic activity was observed in the presence of 5 mM Mg(2+) at pH 7.4. Enzyme activity was completely inhibited by EDTA. PMID- 10790185 TI - CD'99 7th international conference on circular dichroism PMID- 10790186 TI - Determination of absolute configurations and conformations of organic compounds by theoretical calculations of CD spectra AB - The usefulness and limitations of the CNDO/S method for calculations of rotational strengths of inherently chiral chromophores are illustrated with examples from bridged biphenyls. The more empirical Schellman matrix method is employed to calculate CD spectra of compounds containing two chirally disposed chromophores. Analysis of the CD spectra of a compound containing two interacting thioamide groups gives detailed conformational information. Analysis of the CD spectra of two closely analogous 1,2,3, 4-tetrahydro-2-aryl-N-formyl-pyridines with spiro structure shows how a small structural change can lead to the appearance of a nearly mirror image CD spectrum with retained configuration at the spiro atom. Copyright 2000 Wiley-Liss, Inc. PMID- 10790187 TI - Determination of the structure of chiral molecules using ab initio vibrational circular dichroism spectroscopy. AB - We discuss the theoretical prediction of vibrational circular dichroism (VCD) spectra using ab initio density functional theory (DFT) and the application of this methodology to the determination of the absolute configurations and conformations of chiral molecules. PMID- 10790188 TI - Theoretical and experimental studies for the interpretation of vibrational circular dichroism spectra in the CH-stretching overtone region AB - Two theoretical models for the interpretation of the existing data of CH stretching overtones' vibrational circular dichroism data are presented. The first model is based on the quantum mechanical Van Vleck contact transformation theory and is applied to the full vibrational problem, the second is based on classical trajectories calculations, by which we treat a simplified three-degrees of freedom Hamiltonian. The latter allows one to derive a qualitative but efficacious picture of the behavior of coupled anharmonic oscillators. In this framework, we analyze the Poincare Surfaces of Section, and calculate the Fourier Cross Spectra of coupled CH-stretchings. Values for the harmonic frequencies and anharmonicities are derived from absorption spectra in the near infrared on partially deuterated compounds. The effect of large amplitude, low-frequency puckering or twisting modes on the ensemble of coupled CH-stretching is taken into account. Copyright 2000 Wiley-Liss, Inc. PMID- 10790189 TI - Vibrational circular dichroism of tetraphenylporphyrin in peptide complexes? A computational study. AB - The Raman and absorption spectra of tetraphenylporphyrin (TPP) were calculated and compared to experiment. The computation was based on the harmonic molecular force field and electric tensors obtained ab initio at the BPW91/6-31G* level. Good agreement was found between experimental and calculated frequencies and intensities. In order to estimate whether induced optical activity in chiral complexes interferes with the signal of peptide vibrations, the vibrational circular dichroism (VCD) spectra of TPP were simulated. The magnetic field perturbation theory (MFP) and the gauge-invariant atomic orbitals (GIAO) were used for the simulation. Such spectra were compared to theoretical VCD intensities of a model tripeptide as well to experimental spectra of a complex of the peptide and tetrakis(p-sulfonatophenyl)porphyrin (TSPP). No significant contribution to VCD signal from the TPP residue was found in experimental spectra. Thus, possible peptide conformational changes occurring during the complexation can be monitored directly in the amide I frequency region. PMID- 10790190 TI - Measurements of concentration dependence and enantiomeric purity of terpene solutions as a test of a new commercial VCD spectrometer AB - Vibrational circular dichroism (VCD) spectra of (+)-alpha-pinene solutions in carbon tetrachloride have been measured in the range of volume fractions 5-100% (v/v) in the mid-infrared region. The concentration dependence measured was statistically analyzed with the aim of obtaining a reliable correlation between the VCD band areas and the concentrations of individual enantiomers. The quality of the spectra was estimated by means of noise spectra which were defined as half the difference of the two following blocks of scans. In addition to this, the enantiomeric purity was studied. This study was carried out for both (+)- and (-) alpha-pinene enantiomers in the range of the percent enantiomeric excess in the interval 10-100%. The relationship between VCD intensity and enantiomeric purity was determined by least-square regression and statistically evaluated. All measurements performed in this study were intended as a basic tool for testing of a new commercial VCD setup from Bruker. Copyright 2000 Wiley-Liss, Inc. PMID- 10790191 TI - Relative steric size of SCH(3), OCH(3), and CH(3) groups from circular dichroism measurements. AB - The relative steric size of methyl, methoxy, and methylthio groups was determined from circular dichroism (CD) spectroscopy using a sensitive system based on the bilirubin model. In the cyclohexane model, equatorial vs. axial orientation and conformational analysis led to quantitative measurements of orientation preference or steric demand: conformational A-values CH(3) > SCH(3) > OCH(3). A more sterically demanding model for assessing group size has been found in bilirubin analogs, which are yellow pigments that adopt a ridge-tile shape stabilized by a matrix of intramolecular hydrogen bonds. Optically active bilirubins have been shown to exhibit intense bisignate CD Cotton effects from exciton coupling of their two dipyrrinone chromophores held in either of two enantiomeric ridge-tile conformations. Interconversion of these M and P conformational enantiomers of helical chirality is rapid at room temperature but may be displaced toward either enantiomer by intramolecular nonbonded steric interactions that arise when substituents are introduced at equivalent sterically demanding sites, viz., the alpha or beta carbons of the pigment's propionic acid chains. Such substituents shift the conformational equilibrium toward the M or the P-chirality conformer, depending only on the S or R stereochemistry at the alpha and beta sites, and the resulting exciton CD for the approximately 430 nm transition(s) was used to evaluate the relative steric size, SCH(3) > CH(3) > OCH(3). PMID- 10790192 TI - Contribution of alpha140Tyr and beta37Trp to the near-UV CD spectra on quaternary structure transition of human hemoglobin A. AB - The CD band of human adult hemoglobin (Hb A) at 280 approximately 290 nm shows a pronounced change from a small positive band to a definite negative band on the oxy (R) to deoxy (T) structural transition. This change has been suggested to be due to environmental alteration of Tyrs (alpha42, alpha140, and beta145) or beta37 Trp residues located at the alpha1beta2 subunit interface by deoxygenation. In order to evaluate contributions of alpha140Tyr and beta37Trp to this change of CD band, we compared the CD spectra of two mutant Hbs, Hb Rouen (alpha140Tyr-->His) and Hb Hirose (beta37Trp-->Ser) with those of Hb A. Both mutant Hbs gave a distinct, but smaller negative CD band at 287nm in the deoxy form than that of deoxyHb A. Contributions of alpha140Tyr and beta37Trp to the negative band at the 280 approximately 290 nm region were estimated from difference spectra to be 30% and 26%, respectively. These results indicate that the other aromatic amino acid residues, alpha42Tyr and beta145Tyr, at the alpha1beta2 interface, are also responsible for the change of the CD band upon the R-->T transition of Hb A. PMID- 10790193 TI - DNA structure control by polycationic species: polyamine, cobalt ammines, and di metallo transition metal chelates. AB - Many polycationic species bind to DNA and induce structural changes. The work reported here is the first phase of a program whose long-term aim is to create a class of simple and inexpensive sequence-selective compounds that will enable enhanced DNA structure control for a wide range of applications. Three classes of molecule have been included in this work: the polyamine spermine (charge: 4(+)) and spermidine (charge: 3(+)) (which are known to induce a wide range of DNA conformational changes but whose binding modes are still not well understood); cobalt (III) amine transition metal complexes as potential polyamine mimics and [Fe(H(2)O)(6)](3+); and the first member of a new class of di-metallo tris chelated cylinders of helical structure (charge 4(+)). Temperature-dependent absorption, circular dichroism, linear dichroism, gel electrophoresis, and molecular modeling data are presented. The cobalt amines prove to be effective polyamine mimics, although their binding appears to be restricted to backbone and major groove. All the ligands stabilize the DNA, but the 4(+) di-iron tris chelate does so comparatively weakly and seems to have a preference for single stranded DNA. All the molecules studied bend the DNA, with the di-iron tris chelate having a particularly dramatic effect even at very low drug load. PMID- 10790195 TI - Conformational study by CD of chirally tethered naphthalene moieties: toward an understanding of the asymmetric intramolecular coupling reaction? AB - The carbonate (R,R)-1 and the diester (R,R)-2, precursors of optically active 1,1'-binaphthyl derivatives through asymmetric intramolecular coupling, were prepared and fully characterized. The absorption and CD spectra, together with the cholesteric induction measurement of (R,R)-1, indicate that it assumes a conformation in which the naphthalene rings are M-skewed. Since the intramolecular coupling of (R,R)-1 affords a P-twisted (S) binaphthyl derivative, a change of twist from M to P must occur during the reaction. Moreover, this conformation allows the coupling to occur along only one pathway, thus explaining the high stereoselectivity. The CD analysis of (R,R)-2 indicates that it assumes a rigid conformation as well, with the naphthalene rings fixed relative to each other. However, in this compound the naphthalene moieties can give rise to a coupling reaction following two different pathways, leading to oppositely configured binaphthyls with almost the same probability so that low stereoselectivity results. Copyright 2000 Wiley-Liss, Inc. PMID- 10790194 TI - Porphyrins and metalloporphyrins: versatile circular dichroic reporter groups for structural studies. AB - During the last few years, porphyrins and metalloporphyrins have attracted widespread attention as chromophores for studies in circular dichroism (CD), an indispensable chiroptical tool for monitoring chiral interactions. This review summarizes the multifaceted properties of porphyrins and metalloporphyrins, powerful CD chromophores that are characterized by their intense and red-shifted Soret band, propensity to undergo pi-pi stacking, facile incorporation of metals, and ease in varying solubility. Such attributes make porphyrins one of the most attractive and sensitive chromophores used in CD studies. They offer possibilities for studying the stereochemistry of chiral porphyrin assemblies, large organic molecules, biopolymers, and compounds available in miniscule quantities. The tendency of porphyrins to undergo pi-pi stacking and zinc porphyrins to coordinate with amines enable the CD exciton chirality method to be extended to configurational assignments of flexible compounds containing only one stereogenic center. Various artificial porphyrin receptors have been synthesized for the recognition of biologically important chiral guests such as carbohydrates, amino acids, and their derivatives. The induced CD of the host porphyrin Soret band reflects the identity of guests and binding modes of host/guest complexation with high sensitivity. PMID- 10790196 TI - Chirality of aromatic bis-imides from their circular dichroism spectra AB - It is demonstrated that chirality of molecules composed of 1,2,4, 5 benzenetetracarboxydiimide (pyromellitic diimide) or 1,4,5, 8 naphthalenetetracarboxydiimide units is reflected by their exciton Cotton effects. The analysis is based on the calculated (ZINDO/S) excited states of the model diimide chromophores 1a and 2a. Rotation of the diimide chromophores around the C-N bond in diimides 3-5 is evaluated from the dynamic (1)H NMR data. A comparison of chiroptical properties of bis-diimides 3-5 with the CD spectra of bis-imides 6-8 is also presented. Copyright 2000 Wiley-Liss, Inc. PMID- 10790197 TI - CD spectra of solid-state samples AB - Solid-state CD spectroscopy can provide valuable information on conformation, intermolecular interactions, including chirality induction, and enantioselective reactions which occur specifically in the solid state. The technique can, however, suffer from serious artifacts. Some examples of solid-state CD are given and potential artifacts are discussed. Copyright 2000 Wiley-Liss, Inc. PMID- 10790198 TI - Chirality and circular dichroism of oriented molecules and anisotropic phases AB - The results of the "Chirality measurement" circular dichroism along different viewing directions within molecules and phases-the anisotropy of circular dichroism can give suitable information in order to check helicity rules or to analyze the suprastructural chirality of films of organic materials, respectively. These results of the ACD spectroscopy (the CD of anisotropic phases and oriented molecules) show that in an oriented state different information about "chirality" will be gained from different viewing directions. ACD measurements of alpha,beta-unsaturated ketosteroids, TADDOLes and binaphthol derivatives were analyzed. The chiral induction of cholesteric phases, the helical twisting power, is introduced as another chirality measurement which may possibly be a new relative method for the determination of the absolute configuration. Copyright 2000 Wiley-Liss, Inc. PMID- 10790199 TI - Synthesis of the rac-6-hydroxy-3-methoxycyclohex-2-enone and resolution through recrystallization of its diastereomeric N-tosyl-(S)-proline esters AB - The synthesis and resolution of rac-6-hydroxy-3-methoxycyclohex-2-enone, by fractional crystallization of diastereomeric esters of N-protected (S)-proline was shown to be an effective method for preparation of both pure diastereomers (>95% d.e., according to (1)H NMR data). The absolute configuration of the asymmetric center in cyclohexene ring was determined based on the inversed octant rule for the sign of circular dichroism for these cyclohexenones (n-pi* transition band at approximately 300 nm) and further substantiated by chemical conversion of the (-)-(6R,2'S)-diastereomer into the known (+)-(R)-4 hydroxycyclohex-2-enone. Copyright 2000 Wiley-Liss, Inc. PMID- 10790200 TI - An integrating sphere to measure CD from difficult samples AB - Integrating spheres are widely used with UV-Vis and occasionally with infrared spectrophotometers to measure different types of samples, either in transmission mode (scattered transmission accessories) or in total/diffuse reflectance mode. We built a prototype sphere of the demountable type, which fits easily the sample compartment of a commercial CD spectropolarimeter, requiring neither any alignment nor the use of a dedicated photomultiplier. Samples can be inserted either at the sphere entrance (for scattered transmission mode) or in the center of the sphere (for total reflectance experiments). Selected experimental data are presented to evaluate sphere efficiency, its wavelength range and results with a single sample in different forms. Copyright 2000 Wiley-Liss, Inc. PMID- 10790201 TI - MTM1 mutations in X-linked myotubular myopathy. AB - X-linked myotubular myopathy (XLMTM; MIM# 310400) is a severe congenital muscle disorder caused by mutations in the MTM1 gene. This gene encodes a dual specificity phosphatase named myotubularin, defining a large gene family highly conserved through evolution (which includes the putative anti-phosphatase Sbf1/hMTMR5). We report 29 mutations in novel cases, including 16 mutations not described before. To date, 198 mutations have been identified in unrelated families, accounting for 133 different disease-associated mutations which are widespread throughout the gene. Most point mutations are truncating, but 26% (35/133) are missense mutations affecting residues conserved in the Drosophila ortholog and in the homologous MTMR1 gene. Three recurrent mutations affect 17% of the patients, and a total of 21 different mutations were found in several independent families. The frequency of female carriers appears higher than expected (only 17% are de novo mutations). While most truncating mutations cause the severe and early lethal phenotype, some missense mutations are associated with milder forms and prolonged survival (up to 54 years). PMID- 10790202 TI - Ryanodine receptor mutations in malignant hyperthermia and central core disease. AB - Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle that manifests in response to anesthetic triggering agents. Central core disease (CCD) is a myopathy closely associated with MH. Both MH and CCD are primarily disorders of calcium regulation in skeletal muscle. The ryanodine receptor (RYR1) gene encodes the key channel which mediates calcium release in skeletal muscle during excitation-contraction coupling, and mutations in this gene are considered to account for susceptibility to MH (MHS) in more than 50% of cases and in the majority of CCD cases. To date, 22 missense mutations in the 15,117 bp coding region of the RYR1 cDNA have been found to segregate with the MHS trait, while a much smaller number of these mutations is associated with CCD. The majority of RYR1 mutations appear to be clustered in the N-terminal amino acid residues 35 614 (MH/CCD region 1) and the centrally located residues 2163-2458 (MH/CCD region 2). The only mutation identified outside of these regions to date is a single mutation associated with a severe form of CCD in the highly conserved C-terminus of the gene. All of the RYR1 mutations result in amino acid substitutions in the myoplasmic portion of the protein, with the exception of the mutation in the C terminus, which resides in the lumenal/transmembrane region. Functional analysis shows that MHS and CCD mutations produce RYR1 abnormalities that alter the channel kinetics for calcium inactivation and make the channel hyper- and hyposensitive to activating and inactivating ligands, respectively. The likely deciding factors in determining whether a particular RYR1 mutation results in MHS alone or MHS and CCD are: sensitivity of the RYR1 mutant proteins to agonists; the level of abnormal channel-gating caused by the mutation; the consequential decrease in the size of the releasable calcium store and increase in resting concentration of calcium; and the level of compensation achieved by the muscle with respect to maintaining calcium homeostasis. From a diagnostic point of view, the ultimate goal of development of a simple non-invasive test for routine diagnosis of MHS remains elusive. Attainment of this goal will require further detailed molecular genetic investigations aimed at solving heterogeneity and discordance issues in MHS; new initiatives aimed at identifying modulating factors that influence the penetrance of clinical MH in MHS individuals; and detailed studies aimed at describing the full epidemiological picture of in vitro responses of muscle to agents used in diagnosis of MH susceptibility. PMID- 10790203 TI - RET and GDNF gene scanning in Hirschsprung patients using two dual denaturing gel systems. AB - Hirschsprung disease (HSCR) is a congenital disorder characterised by intestinal obstruction due to an absence of intramural ganglia along variable lengths of the intestine. RET is the major gene involved in HSCR. Mutations in the GDNF gene, and encoding one of the RET ligands, either alone or in combination with RET mutations, can also cause HSCR, as can mutations in four other genes (EDN3, EDNRB, ECE1, and SOX10). The rare mutations in the latter four genes, however, are more or less restricted to HSCR associated with specific phenotypes. We have developed a novel comprehensive mutation detection system to analyse all but three amplicons of the RET and GDNF genes, based on denaturing gradient gel electrophoresis. We make use of two urea-formamide gradients on top of each other, allowing mutation detection over a broad range of melting temperatures. For the three remaining (GC-rich) PCR fragments we use a combination of DGGE and constant denaturing gel electrophoresis (CDGE). These two dual gel systems substantially facilitate mutation scanning of RET and GDNF, and may also serve as a model to develop mutation detection systems for other disease genes. In a screening of 95 HSCR patients, RET mutations were found in nine out of 17 familial cases (53%), all containing long segment HSCR. In 11 of 78 sporadic cases (14%), none had long segment HSCR. Only one GDNF mutation was found, in a sporadic case. PMID- 10790205 TI - Russian mutational spectrum differs from that of their Western counterparts. AB - It has been previously noted that the hypoxanthine-guanine phosphoribosyltransferase (HPRT) mutant frequency of Russian subjects is significantly higher than age-matched Western counterparts. To further explore this difference, approximately 100 mutants collected from Russian twins reported in a previous study have been sequenced and compared to an aged-matched Western mutant dataset. The mutational spectrum of the Russian subjects was significantly different (Adams and Skopek Monte Carlo test, P = 0. 004). Curiously, this younger Russian spectrum resembles that recovered from older individuals in the West. Specifically, A:T-->C:G transversions are significantly over-represented (Fisher's Exact test, P = 0.003) in the twin spectrum as compared to the young (age A(S22X). Patient 1 was homozygous for a complex mutant allele containing two alterations, including a 194C>T transition (P65L) and 739A>C transversion (K247Q); in the case of P65L, the amino acid change does not reduce enzyme activity. However, the nucleotide change resulted in exon 3 skipping, whereas the latter K247Q mutation had a drastic effect on enzyme activity. We verified these events by in vivo splicing experiments and transient expression analysis of mutant cDNAs. The P65L mutation locates 11 bases upstream of a splice donor site of intron 3. This is an example of an exonic mutation which affects exon-splicing. PMID- 10790206 TI - Novel mutations in 13 probands with galactokinase deficiency. AB - Galactokinase is an essential enzyme in the metabolism of galactose. Patients with deficiencies in galactokinase exhibit early-onset cataracts. We examined the sequence of the human galactokinase gene (GK1) from 13 patients exhibiting galactokinase deficiency and identified 12 novel mutations. One of the mutations occurred in six of the 13 probands examined, and the remaining 11 were unique mutations. Expression of each of the mutant GK1 genes in Xenopus oocytes resulted in very low galactokinase activity levels. These results provide important information regarding the types of GK1 mutations that occur in the human population. PMID- 10790207 TI - Mutational analysis of ATP7B and genotype-phenotype correlation in Japanese with Wilson's disease. AB - The gene ATP7B responsible for Wilson's disease (WD) produces a protein which is predicted to be a copper-binding P-type ATPase, homologous to the Menkes disease gene (ATP7A). Various mutations of ATP7B have been identified. This study aimed to detect disease-causing mutations, to clarify their frequency and distribution, to determine whether genotype correlates with phenotype, and to determine the rate of abnormal findings in heterozygotes for the WD gene. We analyzed 41 unrelated Japanese WD families, including 47 patients. Twenty-one mutations, including nine novel ones, were identified. 2871delC (15.9%), 1708-5T-->G (11. 0%), and Arg778Leu (13.4%) were the most common mutations. 2871delC was detected mainly in eastern Japan and 1708-5T-->G in western Japan. The homozygotes for the 1708-5T-->G, 2871delC, or Arg778Leu mutations did not show a correlation with their phenotypes. Ceruloplasmin and copper levels were abnormally low in 28.6% and 35. 0% of heterozygotes, respectively. When patients and their families are screened for WD, a high rate of abnormal laboratory data in heterozygotes must be taken into account. PMID- 10790208 TI - Optimization of a simple and rapid single-strand conformation analysis for detection of mutations in the PROS1 gene: identification of seven novel mutations and three novel, apparently neutral, variants. AB - Anticoagulant protein S (PS) deficiency is a known risk factor for thrombophilia. The structure and high allelic heterogeneity of the PS gene (PROS1), together with the presence of a 97% homologous pseudogene, complicates PROS1 analysis. We have optimized a simple, fast, and non-isotopic Single-Strand Conformation Analysis (SSCA or SSCP) method for PROS1 mutation detection. This is accomplished through the analysis of the single-stranded and heteroduplex DNA fragments corresponding to 15 PCR segments that include part of the 5'-upstream region and the 15 PROS1 exons with their intron boundaries. To standardize the method, 13 known PROS1 mutations or allele variants in 10 different fragments were analyzed under different electrophoretic conditions. The results indicated that, using a combination of two different electrophoretic settings, all the allele variants could be detected as a single-strand band shift and/or by the presence of a heteroduplex. This method was used to analyze the PROS1 gene in 31 propositi with different types of PS deficiency and thrombosis. Ten different cosegregating mutations, seven of which are novel (143C->G, L-27H, G96X, M599T, P626L, 1418delA, and 1877delT), were identified in the five families suffering from type I or quantitative PS deficiency and in four of the nine families with coexistence of type I and type III phenotypes. No clearly co-segregating PROS1 mutations were identified in any of the 17 type III propositi analyzed, although eight of them were heterozygotes for the uncommon P460 allele of the S/P460 variant. Furthermore, five apparently neutral allelic variants, three of which are novel ( 296C->T, 182G->C and T57S), were identified in a normal control, two type I/III and two type III PS-deficient pedigrees. PMID- 10790211 TI - Mutations in the human TWIST gene AB - The authors wish to correct a mistake in the amino acid change stemming from a 490C>T nucleotide change. In this article, the nucleotide change is stated to lead to a Q165X amino acid change. The 490C>T nucleotide change should actually cause a Q164X amino acid change. PMID- 10790209 TI - Detection of novel NF2 mutations by an RNA mismatch cleavage method. AB - Mutations in the neurofibromatosis type 2 gene (NF2) cause benign nervous system tumors. Common methods for detecting NF2 mutations (such as single stranded conformational polymorphism analysis and denaturing gradient gel electrophoresis) are laborious and time-consuming. We adapted and improved a commercial assay, the Non-Isotopic RNase Cleavage Assay (NIRCAtrade mark, Ambion, Austin, TX) for rapid, non-isotopic, high-sensitivity screening for NF2 mutations in tumors. We improved the assay by: 1) extending the typical NIRCAtrade mark template size of < 500 bp to 1.3 kb without decreasing detection efficiency; 2) modifying the transcription step of the original protocol so that transcription of PCR products was increased by up to 50%; 3) optimizing the combination of cleavage enzymes and reaction time. With these modifications, mutations were found in 15 of 20 patients (75%) using NIRCAtrade mark. Seven of the point mutations detected (two nonsense, two missense, and three splice-site) are novel. All mutations were confirmed by direct sequencing and no mutations were found using direct sequencing in patients that were negative by NIRCAtrade mark. The 75% NF2 mutation detection rate using this design is similar to detection rates in tumors using other mutation detection methods. PMID- 10790212 TI - Molecular analysis of the hydroxymethylbilane synthase (HMBS) gene in Italian patients with acute intermittent porphyria: report of four novel mutations. AB - Acute intermittent porphyria (AIP) is an autosomal disorder caused by molecular abnormalities in the hydroxymethylbilane synthase (HMBS) gene coding for the third enzyme in the heme biosynthetic pathway. So far, more than 160 different mutations responsible for AIP have been identified in this gene. We have now identified seven mutations in eight unrelated Italian patients with AIP: two splicing defects (IVS7+2T-->C, 612G-->T), three small deletions (308-309delTG, 730-731delCT, 182delA) and two missense mutations (134C-->A, 541C-->T). The splicing defects were responsible for activation of splicing cryptic sites respectively within intron 7 (15 bp insertion) and exon 10 (9 bp deletion). The small deletions resulted in frameshifts leading to the formation of premature stop codons. The 134C-->A and 541C-->T mutations caused the formation of stop codons likely to be responsible for drastic disruption of the HMBS structure (Ser45Ter, Gln181Ter). This is the first molecular study in AIP patients of Italian origin leading to the identification of four new mutations and three molecular defects that have already been described. PMID- 10790214 TI - Characterization of the CYP21 gene 5' flanking region in patients affected by 21 OH deficiency. AB - In order to test the hypothesis that mutations in the 5' non-coding region of CYP21 gene could contribute to the various spectrum of disease presentation due to 21-OH deficiency, the 400bp nucleotide sequence upstream of the ATG codon of CYP21 gene has been characterized in 28 CAH patients who have previously been genotyped by screening for the ten most frequent CYP21 mutations. Six specific sequence variations (-4C-->T, -73C-->T, -295T-->C, -294A-->C, -283A-->G, -281T- >G) have been identified in this region of CYP21 gene in 3 out of 28 21-OH deficient patients for whom the coding region mutations have been previously identified. Three of these mutations, -295T-->C, -294A-->C, -283A-->G, are apparently generated by a gene-conversion event, thus giving first evidence that this mechanism also applies to the 5' untranslated region of CYP21 gene in 21-OH deficiency. Four other sequence changes, identified at nucleotide position -279, 331, -350 and -353, could be referred to as normal since they are present also in healthy subjects. It may not be excluded that some of the newly-identified single nucleotide changes in the regulatory region could have a modulatory effect on the CYP21 gene transcriptional activity thus affecting the clinical outcome. PMID- 10790213 TI - Founder BRCA1 mutations and two novel germline BRCA2 mutations in breast and/or ovarian cancer families from North-Eastern Poland. AB - Germline mutations in the BRCA1 and BRCA2 genes account for the majority of high risk breast/ovarian cancer families, depending on the population studied. Previously, BRCA1 mutations were described in women from Western Poland. To further characterize the spectrum of BRCA1 mutations and the impact of BRCA2 mutations in Poland, we have analyzed 25 high-risk breast and/or ovarian cancer families from North-Eastern Poland for mutations in all coding exons of the BRCA1 and BRCA2 genes, using combined heteroduplex analysis/SSCP followed by direct DNA sequence analysis. Out of 25 probands a total of five (20%) carried three recurrent BRCA1 mutations (300T>G, 3819del5, 5382insC). The 300T>G mutation accounted for 60% (3/5) of BRCA1 mutations and allelotyping suggested a common founder of this mutation. No unique mutations were found. In addition, we identified three BRCA2 (12%) mutations, one recurrent 4075delGT, and two novel frameshift mutations, 7327ins/dupl19 and 9068delA. We conclude that 30% of high risk families from North-Eastern Poland may be due to recurrent BRCA1 and unique BRCA2 mutations. Intriguingly, the BRCA1 mutation spectrum seems to be different within subregions of Poland. PMID- 10790215 TI - Identification of seven novel SNPS (five nucleotide and two amino acid substitutions) in the connexin31 (GJB3) gene. AB - Connexin31 (GJB3) has been associated with hearing impairment and erythrokeratodermia variabilis. We have analyzed this gene in samples from patients with peripheral neuropathies, deafness and controls and have found several single nucleotide polymorphisms (SNPs). In the noncoding exon 1 of GJB3 two small deletions, 581del2 and 632del4 (GenBank accession number AF052692), were found at frequencies of 30% and 14%, respectively. In exon 2 we found two amino acid changes, R32W (1227C-T) and V200I (1731G-A), and three nucleotide variants not affecting the amino acid sequence, 1610G-A, 1700C-T and 1931C-T. Most of these changes were found at similar frequencies in patients with deafness, patients with peripheral neuropathies and control subjects. V200I, 1700C-T and 1610G-A were found associated in three unrelated patients with deafness and in a fourth patient with peripheral neuropathy, but were not detected in control subjects. PMID- 10790217 TI - Detection of point mutations of BCL10 gene in hepatocellular carcinoma tissues: report of 46 cases. AB - BCL10 was found to have truncated mutations at a high frequency in MALT (mucosa associated lymphoid tissue) B cell lymphomas. We examined the mutations of BCL10 gene in human primary liver cancer using non-isotopic PCR-SSCP. Three exons were examined in both cancer and non-HCC adjacent liver tissues. For each exon, six PCR products with abnormal bands were sequenced to verify those mutations. 56.5% samples were revealed a C to G mutation at position 5744 (g5744C>G) of the first exon of BCL10 gene; 54.3% samples were revealed a T deletion mutation at position 11311 (g11311delT) of the second exon of BCL10 gene; 45.7% samples were revealed a C to T mutation at position 14116 (g14116C>T) of the third exon of BCL10 gene. Similar mutation types were found in tumor-adjacent tissues at a lower frequency. The single base changes result in a truncated BCL10 protein expression. Serum alpha-fetoprotein (AFP) level, the tumor size had no significant relationship with BCL10 mutation. PMID- 10790216 TI - Identification of three novel mutations (E196K, V203I, E211Q) in the prion protein gene (PRNP) in inherited prion diseases with Creutzfeldt-Jakob disease phenotype. AB - Inherited prion diseases are characterized by mutations in the PRNP gene encoding the prion protein (PrP). As the other sporadic or infectious prion disease forms, they are almost all characterized by the accumulation in the brain of an abnormal misfolded form of the patient's PrP. Brain extracts can often transmit the disease once inoculated in a recipient animal. Inherited prion diseases with Creutzfeldt-Jakob disease (CJD) phenotype are autosomal forms, although sporadic cases have been reported. We report three novel mutations of the PRNP gene in unrelated patients with clinical and histopathologic features of CJD. The three mutations were missense: c635G>A (E196K), c656G>A (V203I) and c680G>C (E211Q). Familial history of neurologic disorders was evidenced for patients carrying the E196K and E211Q mutations. E196K would be predicted to have more severe effects on protein stability than V203I and E211Q. These mutations expand the spectrum of mutations in PRNP and reduce the proportion of CJD patients in whom genetic alterations have not been found. PMID- 10790218 TI - Analysis of the human KCNH2(HERG) gene: identification and characterization of a novel mutation Y667X associated with long QT syndrome and a non-pathological 9 bp insertion. AB - Long QT (LQT) syndrome is a potentially life-threatening disorder, characterized by a distinct cardiac arrhythmia known as torsades de pointes. Mutations within a number of genes linked to the familial form, including that coding for a cardiac potassium channel called KCNH2 (HERG), have been described based on the characterized genomic organization. A standardized method was developed to screen the entire gene for gene variants. We report a single base pair substitution, introducing a premature STOP codon at codon 667 of the gene in a healthy individual with an extended QTc interval (460 msec). In vitro expression of the codon Y667X variant in Xenopus oocyte suggests that the autosomal dominant variant does not function in a dominant/negative manner and cannot co-assemble to form a channel, resulting in a reduction of the KCNH2 current, and an extension of the QT interval. This indicates that pathogenic LQT gene variants exist in the apparently normal population, the prognosis and clinical consequences of which remain to be determined. The assays described should facilitate future studies into this area. PMID- 10790219 TI - Mutation analysis in 36 unrelated Spanish subjects with familial hypercholesterolemia: identification of 3 novel mutations in the LDL receptor gene. AB - We used the single strand conformation polymorphism (SSCP) method to investigate 36 apparently unrelated Spanish patients with familial hypercholesterolemia (FH) for mutations in the promoter region and the 18 exons and their flanking intron sequences of the low density lipoprotein receptor (LDLR) gene. Nineteen aberrant SSCP patterns were found, and the underlying mutations were characterized by DNA sequencing. In addition, we tested all patients for the presence of mutations in the gene coding for apolipoprotein B (apo B). Five missense mutations (Q71E, S156L, E256K, N543H and T705I), four nonsense mutations (W(-18)X, E10X, Q133X and C255X), six frameshift mutations (211delG, 518delG, 1045delC, 2085del19, 2207insT and 2393del9) and five splicing mutations (313+1G->C, 1061-8T->C, 1845+1G->C, 2140+5G->A and 2390-1G->C) were identified in the LDLR gene. In total, we detected 20 mutations, 3 of which, designated 1045delC, 1845+1G->C and 2207insT, have not been previously described. Seven patients were found to carry two different mutations in the same allele: W(-18)X and E256K (one patient), Q71E and 313+1G->C (two patients), 1061-8T->C and T705I (two patients), 518delG and 2140+5G->A (one patient) and N543H and 2393del9 (one patient). As we expected, there is a broad spectrum of mutations in the LDLR gene, given the genetic heterogeneity of the Spanish population. PMID- 10790220 TI - Severe cystic fibrosis in a Japanese girl caused by two novel CFTR (ABCC7) gene mutations: M152R and 1540del10. PMID- 10790222 TI - A novel missense mutation (D110E) in exon 4 of CFTR (ABCC7) in a CF infant presenting with hypochloraemic metabolic alkalosis. PMID- 10790221 TI - A new BRCA1 germline mutation (E879X) in a Malaysian breast cancer patient of Chinese descent. PMID- 10790223 TI - Two novel polymorphisms, c1086T>C and c1798C>T, in the MADH4/DPC4 gene. PMID- 10790224 TI - Identification of a IVS4-58delATG polymorphism in the human phosphomannomutase 2 (PMM2) gene. PMID- 10790225 TI - A novel missense mutation (Y89C) in exon 3 of the CFTR (ABCC7) gene in a young male. PMID- 10790227 TI - Three novel somatic mutations in the NF2 tumor suppressor gene [g816T>A; g1159A>G; gIVS11-1G>T]. PMID- 10790226 TI - A new polymorphism (N21D) in the exon 2 of the human MDR1 gene encoding the P glycoprotein. PMID- 10790229 TI - Novel intronic polymorphism (+1675G/A) in the human angiotensin II subtype 2 receptor gene. PMID- 10790228 TI - Two novel mutations in the WASP gene in Wiskott-Aldrich patients of Chile origin: W64R and A124E. PMID- 10790230 TI - E-cadherin expression is a sensitive and specific method for detection of carcinoma cells in fluid specimens. AB - The distinction between reactive mesothelial cells and carcinoma in pleural, peritoneal, and pericardial fluids is often difficult. We and others previously showed that E-cadherin, an epithelial-specific adhesion protein, can be useful for this distinction. In this study we tested the sensitivity and specificity of E-cadherin compared to, and in combination with, conventional cytology for assessment of carcinoma in fluids. Cytyc ThinPreptrade mark slides (Marlborough, MA) from 102 sequential fluids were evaluated for E-cadherin expression by routine immunologic techniques. No evidence of E-cadherin staining was seen in 71 cases, while 31 showed unequivocally positive staining. Sensitivity and specificity were independently determined for E-cadherin alone (72% and 97%, respectively), cytomorphology alone (62% and 100%, respectively), and both together (92% and 100%, respectively). We conclude that assessment of E-cadherin expression has sensitivity and specificity comparable to, or better than, conventional cytomorphology. If both cytomorphology and E-cadherin are used together, a definitive and correct diagnosis could have been made on nearly every case in this study. PMID- 10790231 TI - P53 mutations analysis in benign and malignant breast lesions: using needle rinses from fine-needle aspirations. AB - The fine-needle aspiration (FNA) technique is a widely used method for diagnostic assessment of breast diseases. In the current study we investigated the feasibility of sampling material for genetic studies from the same FNA samples as would be used for breast cytology. After making smears for cytological examination, the needle was rinsed into phosphate-buffered saline (PBS) solution. The material gained was sufficient for a polymerase chain reaction (PCR)-based study. As the FNA samples reflect a broad range of breast diseases, it is possible to study genetic changes at various stages of the neoplastic process. We looked for mutations in the p53 tumor suppressor gene in 198 FNA needle rinses, 42 from carcinomas and 156 from cytologically benign lesions. In the malignant samples, 22% carried mutations in the p53 gene. We also looked for p53 mutations in matching tissue sections from tumors and found the FNA needle rinses to represent the tumor well. In addition, three mutations in cytologically benign lesions were found, but none of these 3 patients were diagnosed with malignant tumors in the time frame of the study. The clinical significance of p53 mutations in benign breast tissue remains to be determined. PMID- 10790232 TI - Utility of blasts with a clear halo around the nucleolus as a predictive indicator for disease progression in patients with myelodysplastic syndromes and aplastic anemia. AB - This study was designed to evaluate the utility of blasts with a clear halo around their nucleoli (BCHN) as a predictive indicator of disease progression in myelodysplastic syndromes (MDSs) and aplastic anemia (AA). Bone marrow aspirates from 75 patients with MDSs and 18 with AA were fixed in 95% ethanol solution or 10% neutral formalin and stained with the Papanicolaou method. BCHNs were detected in 57 of 75 patients with MDSs and in 10 of 18 AA patients. Disease progression was restrictedly observed in 17 patients with MDSs who had BCHNs at onset and in 1 patient with AA. The proportion of BCHNs increased with disease progression in these 16 of 17 patients with MDSs. The presence of BCHNs at onset and the increase in proportion of BCHNs during the clinical course of MDSs were significant indications for predicting disease progression. PMID- 10790233 TI - Pleural fluid cytology: immunocytochemistry usage patterns and significance of nondefinitive diagnoses. AB - There has been little study of how pleural fluids are interpreted in actual practice, including the use of immunocytochemistry and nondefinitive diagnoses. Pleural fluid reports (n = 1,330) from 1991-1997 and the University of Iowa cancer database were retrospectively reviewed to determine the cytologic diagnosis, requisition form history, patient survival, and use of immunocytochemistry. Nondefinitive diagnoses were made in 11.3% of cases. Immunocytochemistry was used in 2.6% of cases and aided in making a definitive diagnosis in 71.9% of cases. For patients with a clinical suspicion of malignancy, the percentages of patients who had a nondefinitive, benign, and malignant diagnosis and died of disease were 81.6%, 94.0%, and 90.6%, respectively. In conclusion, if patients had a history of malignancy and a clinical suspicion of recurrence, patient survival was dismal, regardless of the cytologic diagnosis. Immunocytochemistry was used sparsely but often aided in making a definitive diagnosis. PMID- 10790234 TI - Incidence of apocrine cells in fine-needle aspirates of gynecomastia: a study of 100 cases. AB - While the presence of apocrine cells in fibrocystic conditions of the female breast is a common finding in a fine-needle aspirate (FNA) of the breast, such a finding in gynecomastia has only been reported recently in a single case report. This study was undertaken in 100 cases with adequate cellular samples which had been diagnosed as gynecomastia from aspirates which were obtained using a 22 gauge needle and a 5-ml syringe maintaining negative pressure. The material was collected as needle and syringe washings in 30% ethyl alcohol in physiologic saline and the cytologic preparations were made on 3-microm Schleicher and Schuell filters and were stained by the Papanicolaou method. In 3 of the 100 cases, sheets of apocrine cells were noted in addition to clusters of hyperplastic ductal cells. The findings were further confirmed histologically in cell blocks which were made in the 3 cases. Based on our findings, it was concluded that apocrine cells in gyneco- mastia are found in about 3% of cases, and such a finding should be regarded as nonneoplastic, thus avoiding the need of surgical excision. PMID- 10790235 TI - Fine-needle aspiration of dedifferentiated chondrosarcoma of the larynx. AB - We present a case of dedifferentiated chondrosarcoma of the larynx in which the dedifferentiated component was initially diagnosed by fine-needle aspiration (FNA). The patient was a 74-yr-old man who presented with difficulty breathing and an anterior neck mass. A CT scan demonstrated a 4.5-cm cartilaginous lesion involving the left thyroid cartilage, with an anterior soft-tissue component. Nine years prior, the patient had an incomplete resection of a low-grade chondrosarcoma at the same site. FNA was performed on the current lesion, demonstrating a high-grade spindle-cell sarcoma with a storiform pattern. The cytomorphology together with immunocytochemistry and electron microscopy were diagnostic of malignant fibrous histiocytoma (MFH), and synthesis of the clinical, radiographic, and cytomorphologic features resulted in a diagnosis of dedifferentiated chondrosarcoma. The cytologic diagnosis was histologically confirmed by laryngectomy. Although rare, dedifferentiated chondrosarcoma should be included in the differential diagnosis of high-grade sarcomas of bone and cartilage assessed by FNA. PMID- 10790236 TI - Fine-needle aspiration cytologic diagnosis of intrahepatic biliary papillomatosis (intraductal papillary tumor): report of three cases and comparative study with cholangiocarcinoma. AB - Biliary papillomatosis is a rare tumor of the intrahepatic and extrahepatic biliary tree, and its FNA findings have not been reported. The cytologic features of 3 cases of intrahepatic biliary papillomatosis were studied and compared with 5 cases of cholangiocarcinoma. The distinctive features include: 1) hypercellular smear, 2) very broad and often double-cell layered sheets of ductal columnar epithelium, 3) papillary configuration, 4) preserved honeycomb pattern with even nuclear spacing, and 5) dysplastic but not frankly malignant nuclear features. The constellation of these features is highly characteristic of biliary papillomatosis and helpful in distinguishing it from cholangiocarcinoma and other differential diagnoses. A firm preoperative diagnosis can thus be achieved, allowing better planning in management of this borderline malignant tumor. PMID- 10790237 TI - Fine-needle aspiration cytology of a case of oncocytic adrenocortical carcinoma. AB - We report on the results of fine-needle aspiration cytology of a case of oncocytic adrenocortical carcinoma in a 39-yr-old man. The tumor invaded the inferior vena cava and extended up to the right atrium. Aspirate smears were very cellular and showed a monomorphic population of large polyhedral cells with abundant granular cytoplasm, predominantly distributed singly. Mitotic activity was inconspicuous, and there was no necrosis. Immunohistochemically, the tumor cells were positive for vimentin, cytokeratin, and p53, and negative for synaptophysin, chromogranin, inhibin, and S-100. Ultrastructurally, the cytoplasm of the tumor cells was packed with mitochondria. The patient underwent left radical nephrectomy as well as a combined cardiopulmonary bypass, with atriotomy and resection of the tumor from the right atrium and inferior vena cava. Three months of postoperative follow-up were uneventful. PMID- 10790238 TI - Pemphigus vulgaris of the uterine cervix revisited: case report and review of the literature. AB - Pemphigus vulgaris is an autoimmune disease characterized by acantholytic blisters and erosions involving the oral mucosa, skin, and less frequently other mucosal surfaces. Although the cytology of scrapings from the cutaneous and oral lesions has been well-documented, there are relatively few reports in the literature of the cytologic appearance of pemphigus on cervicovaginal smears. This report documents a case of pemphigus involving the cervix, in which the diagnosis was not known at the time of the cervical smear and biopsy. The cytologic features of this case and those in the literature are described in detail, highlighting the necessity of awareness of the disease and its presentation on cervicovaginal smears, in preventing an overdiagnosis of neoplasia. PMID- 10790239 TI - Fine-needle aspiration cytology of mucous retention cyst of the tongue: distinction from other cystic lesions of the tongue. AB - Mucous retention cyst (MRC) is a common submucosal lesion of the oral cavity that, when deeply seated, simulates a neoplasm. This report describes the fine needle aspiration cytology findings of a lingual MRC of complex architecture and with metaplastic epithelium. In addition, we emphasize its cytologic differential diagnosis from other benign and malignant cystic lesions of the tongue. To the best of our knowledge, this is the first report of aspiration cytology of a complex MRC of the tongue. PMID- 10790240 TI - Fine-needle aspiration cytology of unsuspected metastatic thyroid carcinoma initially presenting in bone: report of three cases and a review of the literature. AB - We present a series of three cases of unsuspected thyroid carcinoma presenting as bony lesions and initially studied by fine-needle aspiration (FNA). This series consists of two cases of follicular carcinoma and a third case of Hurthle cell carcinoma. FNA was performed on all three lesions. In two of these cases, a definitive diagnosis of metastatic thyroid carcinoma was made based on the FNA material. FNA smears of both follicular carcinomas displayed cohesive clusters of atypical round cells with nuclear overlapping and positive immunoperoxidase staining for thyroglobulin. The aspirates of the Hurthle cell carcinoma were composed of sheets and individually scattered oval oncocytic cells with prominent nucleoli. All three cases demonstrated the presence of marginal vacuoles consistent with "flame" cells. To the best of our knowledge, this is the first series discussing the cytologic features and differential diagnosis of unsuspected thyroid carcinoma initially presenting as metastatic bone lesions. PMID- 10790241 TI - Semiquantitative analysis of the cellular preservation quality of Normosol and Carbowax solutions for thyroid fine-needle aspiration specimens. AB - The present study was designed to compare cytomorphologic features and diagnostic value of thyroid fine-needle aspiration (FNA) specimens stored in Saccomano's (Carbowax(R) solution) fixative and Normosol(R) solutions. Equivalent amounts of aspirate material obtained from nonlesional areas of 17 freshly resected thyroidectomy specimens were placed in room temperature Carbowax(R) solution and refrigerated Normosol(R) solution for a time period of up to 60 hr. Millipore(R) filter preparations were made from both fluids for each of the 17 thyroidectomy specimens. Slides were examined by three independent observers who rated the quality of cellular preservation based on a four-part semiquantitative scale. Statistical analysis of the data obtained from the scores of the three observers showed no statistically significant difference in the quality of specimen preservation. Normosol(R) preserved specimens consistently rated superior to Carbowax(R) in nuclear detail, lack of obscuring background artifact, and diagnostic value. These findings, along with a considerably less cost, make refrigerated Normosol(R) an attractive alternative for the short-term storage of FNA specimens placed in fluid. PMID- 10790242 TI - Detection of circulating tumor cells and micrometastases in stage II, III, and IV breast cancer patients utilizing cytology and immunocytochemistry. AB - Evaluation for circulating tumor cells and bone marrow micrometastases has generated considerable interest due to a potential association with disease recurrence and poor prognosis. In this study, we examined bone marrow and apheresis samples from Stage II, III, and IV patients (n 120) enrolled in various clinical breast cancer trials at the National Institutes of Health/National Cancer Institute. For each patient sample, two Diff-Quik-stained cytospins were reviewed for morphology, and approximately 1 x 10(6) cells were analyzed for the expression of cytokeratins using an avidin-biotin immunoperoxidase method. Keratin-positive malignant cells appearing as single cells or in small clusters were detected in bone marrow samples from Stage IV patients only (9/68, 13%) and detected in apheresis samples from both Stage III and IV patients (13/245, 5%). These findings indicate that the combination of cytomorphology with immunocytochemistry can be utilized for the investigation of circulating tumor cells and bone marrow micrometastases, and that positive results appear to correlate with high tumor stage/burden. PMID- 10790243 TI - Aspiration cytology of metanephric adenoma of the kidney. PMID- 10790244 TI - Parental factors affecting respiratory function during the first year of life. AB - In a prospective, longitudinal, population-based cohort study of familial and environmental influences on the development of wheezing respiratory illness in early childhood, we identified infant length, weight, gender, and exposure to maternal cigarette smoking as significant determinants of lung function during the first year of life. A cohort of 237 infants (106 females: 131 males) was evaluated, and 496 lung function measurements were made between the ages of 1-12 months. Respiratory function was assessed using the rapid thoracic compression technique to obtain maximum expiratory flow at functional residual capacity (V'maxFRC). Parental history of asthma and smoking habits during pregnancy were obtained by questionnaire. Data were analyzed using a longitudinal random effects model. Infants with a parental history of asthma and/or in utero passive smoke exposure were compared to a reference group of infants who had no parental history of asthma and in whom neither parent smoked pre- or postnatally. Boys were found to have a consistently lower V'maxFRC (-21.05 mL.s(-1)) throughout the first year of life in comparison to girls (P < 0.05). Maternal smoking during pregnancy was associated with a lower V'maxFRC in both genders in comparison to unexposed infants (P < 0.05). V'maxFRC was unaffected by parental history of asthma. Gender-specific normative equations for V'maxFRC throughout the first year of life were derived for the infant cohort as a whole and also for subgroups of infants, based on parental asthma and smoking history. We conclude that lung function during the first year of life differs between genders and is adversely affected by in utero passive tobacco smoke exposure. Gender-specific predictive equations for V'maxFRC should be used during infancy. PMID- 10790245 TI - Clinical course of postinfectious bronchiolitis obliterans. AB - We performed a prospective observational study to define the clinical course and the prognostic factors of 31 patients with postinfectious bronchiolitis obliterans. All patients presented with an episode of acute bronchiolitis in the first 2 years of life, and respiratory symptoms and signs persisted since then. Other diseases which may cause chronic airflow obstruction were excluded. The patients were followed after their inclusion in the study and the clinical findings were recorded in a standardized questionnaire and form. Repeated chest radiographs and lung perfusion scans were obtained in all 31 patients and semiannual spirometry was performed in 8 older patients. Eight patients had lung biopsies. The clinical course varied in the 31 patients during a mean of 3.5 years of follow-up. The outcome of the patients included clinical remission (22.6%), persistence of respiratory symptoms and signs (67.7%), and death (9.7%). An older age at onset of illness and presence of atopy as suggested by an elevated serum IgE appeared to predispose to a poor prognosis. PMID- 10790247 TI - Dynamic changes in EEG spectra during obstructive apnea in children. AB - Children are less likely to demonstrate EEG arousal during obstructive sleep apnea (OSA) than adults. We hypothesized that changes in spectral EEG characteristics occur during REM-associated OSA in the absence of arousal. Eight snoring children underwent overnight polysomnography. OSA events during REM periods not associated with EEG or behavioral arousal were identified. EEG signals from C3A2 and C4A1 leads corresponding to 1) < or =10-sec epochs preceding OSA (PRE), 2) the obstructed period (OSA), and 3) < or =10-sec epochs following airflow resumption (POST) were subjected to fast Fourier transform (FFT) routines. Seventy-two isolated OSA, and 14 clusters of > 4 OSA events were analyzed. In single OSA, delta OSA amplitude was lower than in PRE (P < 0.01) and in POST (P < 0.001). Furthermore, POST delta amplitude was higher than PRE (P < 0.01). In contrast, in OSA clusters, the dynamic differences in delta amplitude disappeared after the second OSA. Reciprocal increases and decreases occurred for the theta frequency domain during OSA and post-OSA, while sigma and beta frequency power did not change. We conclude that during isolated OSA episodes without arousal, significant decreases in power selectively occur for delta frequency, and are followed by a rebound increase upon termination of apnea. The delta changes are progressively attenuated during repeated OSA. We postulate that delta changes may reflect ongoing adaptations in sleep pressure which are necessary to relieve the respiratory compromise, and may represent subtle evidence for arousal and consequent sleep fragmentation in children with OSAS. PMID- 10790246 TI - Cardiac rhythm disturbances among children with idiopathic congenital central hypoventilation syndrome. AB - The objective of this study was to determine whether subjects with congenital central hypoventilation syndrome (CCHS) had an increased frequency of cardiac arrhythmias and decreased heart rate variability when compared to subjects without a known deficit in control of breathing, and that these abnormalities would be exaggerated by anesthesia. Continuous ambulatory Holter recordings were obtained in patients with CCHS and compared to two otherwise healthy control groups without a deficit in control of breathing: one with an intact airway (n = 11) and a second group with a tracheostomy (n = 6). Holter recordings were obtained before, during (under general anesthesia), and after bronchoscopy. Fourteen children with CCHS (age: 9.3 +/- 4.4 years mean +/- S.D.) were studied, and 7 underwent bronchoscopy. Seventeen control children were studied (age 6.6 +/ 3.6 years): 11 without a tracheostomy, and 6 with a tracheostomy who also underwent bronchoscopy. Maximum heart rate during baseline recording was significantly lower in the CCHS subjects as compared to controls (P = 0.0001). At baseline the difference in the number of arrhythmias/24 hr/subject in all CCHS vs. all control subjects was significant (P = 0.0002); for the subjects who had bronchoscopy, CCHS vs. control, the difference was also significant (P = 0.03). In addition, there was a significant decrease in the number of events/24 hr/subject among the CCHS subjects between baseline and post-bronchoscopy (P = 0.0288). The predominant arrhythmias were sinus bradycardia and transient asystole. The longest asystole in a CCHS subject was 6.50 sec, and in a control subject, 1.42 sec (at baseline the means of the longest asystole were 2.69 +/- 1.4 vs. 1.24 +/- 0.13; P = 0.003 in the CCHS vs. control groups). Other indices of heart rate variability were significantly reduced in the CCHS subjects (P < 0.05). These results substantiate our hypothesis that subjects with CCHS have more arrhythmias than controls, an increased frequency of bradyarrhythmias, and decreased cyclical sinus arrhythmia. PMID- 10790248 TI - Inhaled tobramycin and bronchial hyperactivity in cystic fibrosis. AB - The use of inhaled tobramycin for prophylaxis and treatment of respiratory symptoms in cystic fibrosis (CF) is now widespread. There have been concerns that inhaling the intravenous (I.V.) formulation of tobramycin causes bronchoconstriction. Previous studies using this formulation have either not specified the nebulizing equipment, or studied older, more severely affected patients. This study investigated the incidence of bronchoconstriction with tobramycin inhalation in children with mild to moderate CF. We studied 26 patients between the ages of 7 and 17 years, with mild to moderate CF (20 female). Prior to being placed on prolonged inhaled tobramycin therapy, they underwent a "tobramycin challenge." FEV(1) was measured pre and post challenge. For the test, standard I.V. solution (80 mg/2 mL) diluted with 2 mL of normal saline was nebulized, using the Hudson (Temecula, CA) RCI Updraft II nebulizer. The nebulization lasted 2 min. There was a 3-min "quiet period," following which FEV(1) was measured. A decrease in FEV(1) by at least 10% post-tobramycin inhalation was considered to be a positive test. Results were analyzed using the Pearson Chi-square test. Five of 26 (19%) had a positive reaction to tobramycin. Sixteen of 26 (61.5%) were using salbutamol on a daily basis at the time of testing but not for 48 hr before the challenge, and 16 of 26 (61.5%) had a pre tobramycin FEV(1) of < or =80%. Neither an FEV(1) of <80% (P = 0.93) nor regular use of salbutamol (P = 0. 34) were associated with a positive tobramycin challenge. This study suggests that, while bronchoconstriction does occur, many patients do not exhibit bronchoconstriction in response to the standard I.V. preparation and, as prior work suggests, this may be reduced further by pretreatment with salbutamol. PMID- 10790249 TI - Benefits of music therapy as an adjunct to chest physiotherapy in infants and toddlers with cystic fibrosis. AB - Routine chest physiotherapy (CPT) is an important component of prophylactic therapy in children with cystic fibrosis (CF) and requires a significant commitment of time and energy. It is important, therefore, to establish CPT as a positive experience. In this study, we evaluated the effect of recorded music as an adjunct to CPT. Specifically, we compared the use of newly composed music, familiar music, and the family's usual routine on children's and parents' enjoyment of CPT and the parents' perception of time taken to complete CPT. Enjoyment and perception of time were evaluated via questionnaires designed specifically for this study. Participants were caregivers of one or more children with CF who were aged between 4(1/2) months and 24 months at the commencement of the clinical trial and required CPT on a daily basis. Participants were randomly allocated into control and treatment groups. Control group participants experienced two conditions consecutively: no audiotape (NT; control) and familiar music tape (FT; placebo control). Treatment group participants were given the treatment music tape (TT), which was composed and compiled by a music therapist. After baseline assessment, evaluation occurred at two 6-weekly intervals. Children's enjoyment increased significantly after use of the TT (+1. 25 units) compared to NT (-0.5 units; P = 0.03), as did parents' enjoyment (+1.0 vs. 0.0 units, P = 0.02). Children's enjoyment did not change significantly after use of the FT (+0.75 units) compared to NT (n.s.). Likewise, parents' enjoyment did not change significantly after use of the FT (+1.0 units, n.s.). There was no change in perception of time after use of the TT (-4.5 vs. +0.2 min, n.s.) or the FT (+3.3 min, n.s.). These results indicate that children's and parents' enjoyment of CPT significantly increased after the use of specifically composed and recorded music as an adjunct. We therefore recommend that recorded music, such as that provided in this study, be given to parents to use as an adjunct to CPT when their young children are diagnosed with CF, in order to assist the establishment of a positive routine. PMID- 10790250 TI - Pulmonary renal syndrome in childhood: a report of twenty-one cases and a review of the literature. AB - In adults, the term specific pulmonary renal syndrome describes disorders with pulmonary and glomerular manifestations and includes Wegener's granulomatosis, Goodpasture disease, and systemic lupus erythematosus. Nonspecific pulmonary renal syndrome refers to either pulmonary disease complicating glomerular disease, or glomerular diseases following pulmonary disease. Since little is known regarding pulmonary renal syndrome in childhood, we reviewed the charts of 21 pediatric patients with pulmonary renal syndromes treated by the Department of Pediatrics, University of Bern between 1991 and 1998; we also reviewed the pediatric literature that deals with specific pulmonary renal syndromes. Specific pulmonary renal syndrome was noted in 3 children with systemic vasculitis (Wegener granulomatosis, N = 2; microscopic polyangiitis, N = 1) and 2 with systemic lupus erythematosus. Nonspecific pulmonary renal syndrome was observed in 12 patients with pulmonary edema (N = 9), pulmonary thromboembolism (N = 2), and pulmonary infection (N = 1) complicating the course of a glomerular disease, and in 4 children with a pulmonary disease followed by a glomerular disease. Review of the literature disclosed 52 cases of specific pulmonary renal syndrome other than systemic lupus erythematosus: Wegener granulomatosis (N = 28), Goodpasture disease (N = 13), and Henoch-Schonlein purpura (N = 11). In addition, hemolytic uremic syndrome complicated pneumococcal pneumonia in 32 cases. We conclude that pulmonary renal syndromes need to be looked for in childhood. Apart from Wegener granulomatosis, Goodpasture disease, and systemic lupus erythematosus, Henoch-Schonlein purpura and hemolytic-uremic syndrome occasionally have both pulmonary and renal features. PMID- 10790251 TI - High-percentage lung delivery in children from detergent-treated spacers. AB - Pressurized metered-dose inhalers attached to spacers are now the most common form of delivery of anti-asthma medication in children. However, no reliable data are available of how much drug reaches the lungs in children of different ages. This information is crucial, as it determines the efficacy of therapy. In this study, we present information on the amount of drug reaching the lungs in children from a pressurized metered-dose inhaler attached to a detergent-coated spacer. We studied 18 asthmatic children inhaling radiolabeled salbutamol through detergent treated spacers to minimize electrostatic charge on the spacer wall. Lung deposition was much higher than expected when using detergent-coated spacers. Mean (SD) lung deposition, expressed as a percentage of the total actuated dose (five actuations), was 16.4% (5.5) in younger children inhaling through a small volume spacer, and 28.2% (6.7) and 41.8% (3. 8) in older children inhaling with different breathing patterns through a large volume spacer. These findings have major implications for dosage regimens for inhaled anti-asthma medication in children. Lower doses may be sufficient for adequate drugs delivered through spacers treated for static to achieve a desired clinical response. PMID- 10790252 TI - Recent advances in understanding the pathogenesis of nitrofen-induced congenital diaphragmatic hernia. AB - In this review, we discuss recent advances in the study of the pathogenesis of congenital diaphragmatic hernia (CDH). Much of the research has involved the use of an animal model of CDH in which diaphragmatic defects are produced in fetal rats by administering the herbicide nitrofen to dams during mid-gestation. The animal model is described and the relevance to the human condition is discussed. The data derived from the animal studies are critically assessed in the context of commonly cited hypotheses proposed for the pathogenesis of CDH. Finally, experimental strategies are proposed for systematically examining the normal and pathological formation of the pleuroperitoneal fold. We conclude that a malformation of the primordial diaphragm, the pleuroperitoneal fold, underlies the muscle defects associated with CDH. PMID- 10790253 TI - Vibrio vulnificus infection complicated by acute respiratory distress syndrome in a child with nephrotic syndrome. AB - A 9-year-old girl with nephrotic syndrome visited a local hospital after developing fever, chills, and edematous changes and multiple hemorrhagic bullae on both legs over 2 days. Cultures of blood and an aspirate from the bullae yielded Vibrio vulnificus. The patient was transferred to our hospital because of persistent fever, generalized edema, acute renal failure, and disseminated intravascular coagulopathy. We treated this patient as a V. vulnificus infection complicated with necrotizing fasciitis. With minocycline and ceftazidime combination therapy was instituted. Emergency fasciotomy and continuous peritoneal dialysis were performed. The patient developed acute respiratory distress syndrome (ARDS) during the hospitalization, requiring intubation and mechanical ventilation. She eventually died. The histopathological findings showed diffuse alveolar damage with lobular pneumonitis. Hyaline membranes, composed of proteinaceous exudate and cellular debris, covered the alveolar surfaces. Microscopic examinations of lung could not distinguish the effects of cytolysin from other insults to lungs that occur in ARDS. This report highlights the postmortem pathological findings in V. vulnificus infection in a child with nephrotic syndrome complicated by ARDS. PMID- 10790254 TI - Potential role of high-frequency ventilation in the treatment of severe congenital pleural effusion. AB - Newborns with severe congenital pleural effusions often present with respiratory failure at birth. We describe two premature infants born at 31 and 33 weeks of gestation with bilateral pleural effusions. Both were drained prior to delivery under ultrasound guidance. The first infant had severe bilateral congenital chylothorax with pulmonary hypertension; the second infant had severe nonimmune hydrops fetalis. Both could be adequately oxygenated but failed to respond to conventional mechanical ventilation (CMV) and chest tube drainage, so that CO(2) elimination could not be accomplished. Both infants were successfully treated with high-frequency ventilation (HFV). We suggest that HFV may be of significant value in establishing adequate ventilation in cases of severe congenital pleural effusions. PMID- 10790255 TI - Selected abstracts PMID- 10790256 TI - NAALADase inhibitors: A novel approach to glutamate regulation. PMID- 10790257 TI - Current approaches to multiple parallel synthesis. PMID- 10790258 TI - beta-Sheet breaking peptide might treat prion disease. PMID- 10790259 TI - Obesity: on the eve of a major conceptual revolution. PMID- 10790260 TI - An SOS for HIV. PMID- 10790262 TI - HIV-1 entry - an expanding portal for drug discovery. AB - The advent of highly active antiretroviral therapy (HAART)-combinations of protease and reverse transcriptase inhibitors-provided a potent and clinically effective method of suppressing viral load in HIV-1- infected individuals. However, although initially successful, a broader clinical experience has revealed limitations in this therapeutic regimen, with up to 40% of treated individuals ultimately failing to sustain control over viral replication. Significant advances in understanding the process by which HIV-1 enters host cells have brought into clear focus a target for drug discovery not represented in the current clinical armamentarium. In this article, the mechanism of HIV-1 entry is reviewed in the context of representative antiviral agents that interfere with key steps in this process. PMID- 10790263 TI - Do antibiotics maintain antibiotic resistance? AB - Important human pathogens resistant to antibiotics result from the human use of antibiotics. Does this imply that reducing their usage or removing antibiotics from medicine and agriculture will restore the effectiveness of these drugs? The authors argue that resistance evolution and susceptibility evolution are not, in a sense, just different sides of the same coin. Resistance genes acquire new functions and the initial costs of resistance can evolve into advantages. Decreasing drug use might not replace a fundamental change in drug design to avoid the evolution of resistant, and encourage the evolution of susceptible, microorganisms. PMID- 10790264 TI - Regulation of clinical trials in Europe. AB - Pharmaceutical companies spend billions of dollars every year on carrying out clinical trials for their potential products. Many other trials are funded by charities, government research councils and other bodies. However, within Europe, national differences in approval procedures and the law relating to clinical trials can lead to increased costs and delays, particularly where trials are conducted in different countries. New European legislation is currently under review that is designed to ensure a common level of patient protection and scientific standards whilst reducing the costs and delays that can occur before trials can commence PMID- 10790265 TI - Monitor: molecules. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 10790266 TI - Combinatorial chemistry. PMID- 10790267 TI - High-throughput screening. PMID- 10790268 TI - Track effects of heavy ions in liquid water. AB - The various types of ionizing radiation can have widely differing radiation effects due to their variation in track structure. The effects of track structure on the radiation chemistry of water are particularly important because of the fundamental aspects and the wide range of practical applications. This review gives an overview of how the physics of energy-loss processes are responsible for the geometry of the particle track and thereby the final product yields. The radiation chemical effects are discussed in qualitative terms to show how the fundamental relationship between the physics and chemistry of particle tracks leads to the observed products. Special phenomena at very high rates of energy deposition are also covered. Finally, a discussion on the future research trends is given. PMID- 10790269 TI - Protection of endogenous vitamin E and beta-carotene by aminoguanidine upon oxidation of human low-density lipoproteins by *OH/O(2)*-. AB - This study was designed to evaluate the antioxidant effect of aminoguanidine toward human low-density lipoproteins (LDLs) initiated by oxygenated free radicals (*OH/O(2)*-) generated by gamma radiolysis. Initial radiolytic yields related to the markers of lipid peroxidation [i.e. decrease in endogenous alpha tocopherol and beta-carotene, formation of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes] were determined in 3 g liter(-1) LDLs (expressed as total LDL concentration) in the absence and presence of 10 different concentrations of aminoguanidine (from 0.04 to 5 mmol liter(-1)). Fluorescence and relative electrophoretic mobility of oxidized LDLs were also studied as markers that indirectly reflect the attack of the protein moiety of LDLs (namely apolipoprotein B). Our data clearly showed the inhibitory effect of aminoguanidine on lipid peroxidation induced in LDLs by *OH/O(2)*- in a concentration-dependent manner. This effect probably resulted from a scavenging activity of aminoguanidine toward *OH. In contrast, aminoguanidine did not appear to react significantly with O(2)*-, which resulted in a poor residual lipid peroxidation. Our data led us to determine an optimum [aminoguanidine]/[LDL] ratio ranging from 250 to 500 to obtain the best in vitro protection of LDLs under our experimental conditions. It is also of great interest that aminoguanidine was able to protect endogenous alpha-tocopherol and beta-carotene of LDLs upon *OH/O(2)*(-)-induced oxidation. PMID- 10790270 TI - Relative contribution of bystander and targeted cell killing to the low-dose region of the radiation dose-response curve. AB - Human keratinocytes show a bystander effect when exposed to low doses of low-LET radiation. In this paper, data are presented showing a method of correcting the overall survival curve to enable analysis of the relative contributions of the bystander effect and the effect attributable to direct interaction of the radiation with the target cell. The technique used is to obtain a standard clonogenic survival curve using the assay of Puck and Marcus and, with a different set of flasks containing cloning densities of unirradiated cells, to assay the cell killing caused by medium harvested from 2 x 10(5) cells irradiated with the same doses. The data show that for this human epithelial cell line, doses of 0.01-0. 5 Gy show clonogenic death by the bystander effect only, if maximum potential bystander killing is assumed. The magnitude of the effect is relatively constant, and it appears to saturate at doses in the range of 0.03 0.05 Gy. After doses greater than 0.5 Gy, the curves for clonogenic death are the result of a dose-dependent non-bystander effect and a dose-independent bystander effect. If these particular dose-response effects occur in epithelial cells in vivo, they may have important consequences for therapy and studies of low-dose risk. PMID- 10790272 TI - Induction and repair of DNA double-strand breaks under irradiation and microgravity. AB - The influence of microgravity on induction and repair of double-strand breaks was studied in the yeast mutant rad54-3, which is temperature-conditional for the repair of DNA double-strand breaks. The experiment was performed on the shuttle Atlantis flight STS-84. Cell samples were kept at 0-4 degrees C until they reached orbit, where they were transferred to 22 (permissive temperature for repair) and 37 degrees C (restrictive temperature). They were exposed to graded doses of beta particles from an in-built (63)Ni source during the repair period. After 152 h in microgravity, the radiation exposure was stopped, and the samples were returned to low-temperature conditions, where they remained until final evaluation in the home laboratory. The amount of double-strand breaks remaining was estimated from the differences in survival after plating and incubation at the restrictive temperature. The results show that there is no significant difference for both the induction and the repair of double-strand breaks between microgravity and terrestrial conditions. PMID- 10790271 TI - Induction of DNA single-strand breaks by 131I and 99mTc in human mononuclear blood cells in vitro and extrapolation to the in vivo situation. AB - The radionuclides (131)I and (99m)Tc are frequently used for therapy of benign and malignant thyroid disease ((131)I) and for diagnosis of thyroid and other diseases ((99m)Tc). However, the levels of DNA single-strand breaks (SSBs) induced in cells of patients after administration of (131)I and (99m)Tc are not known. In this study, we measured the number of SSBs per cell induced by (131)I and (99m)Tc in vitro, extrapolated the results to the clinical situation, and assessed their biological relevance by comparing levels of SSBs induced after therapeutic administration of (131)I and (99m)Tc to those induced by endogenous processes or by occupational exposure to genotoxic substances. A linear dose response relationship between the radioactivity concentrations of (131)I and (99m)Tc and SSBs in human mononuclear blood cells (determined by alkaline elution) was obtained after incubation at 4 and 37 degrees C. At 4 degrees C, where almost no repair of SSBs takes place, (131)I and (99m)Tc induced 81 and 7 SSBs per cell per hour/(MBq/ml), respectively. At 37 degrees C, only 20 and 1.6 SSBs per cell per hour/(MBq/ml) were observed after incubation with (131)I and (99m)Tc. To estimate the induction of SSBs in vivo in cells of patients after administration of 3700 MBq (131)I (oral) or 60 MBq (99m)Tc (i.v.), the rates of induction of SSBs obtained in vitro were extrapolated to the concentrations of (131)I and (99m)Tc measured in blood of patients. The total number of SSBs (mean +/- standard deviation) accumulated after oral administration of 3700 MBq (131)I up to 70 h after administration was calculated as 200 +/- 59 SSBs/cell. After administration of 60 MBq (99m)Tc (i.v.), 0.032 +/- 0.009 SSBs per cell (total SSBs up to 2 h after administration) were cumulated. The induction of SSBs by endogenous processes (estimated 2,000 SSBs per cell per hour) and by occupational exposure to genotoxic substances (125-430 SSBs per cell) has been estimated in earlier studies. In conclusion, the frequency of SSBs induced by thyroid diagnosis with 60 MBq (99m)Tc is approximately 5 orders of magnitude smaller than the frequency of spontaneous SSBs and thus is most probably without biological relevance. Since the frequency of induction of SSBs by therapy with (131)I (3700 MBq) is about 6000-fold higher compared to thyroid diagnosis by (99m)Tc, its biological relevance is more difficult to assess. Nevertheless, the number of SSBs induced by therapy with (131)I is substantially lower than that induced by endogenous processes. PMID- 10790273 TI - Assessing exposure to cosmic radiation during long-haul flights. AB - The assessment of exposure to cosmic radiation on board aircraft is one of the concerns of organizations responsible for radiation protection. Cosmic-particle flux increases with altitude and latitude and depends on solar activity. To illustrate the effect of these parameters, exposure has been estimated on several airlines operating subsonic and supersonic aircraft on transatlantic, Siberian and transequatorial routes. Measurements have been made with a tissue-equivalent proportional counter using the microdosimetric technique. This type of system provides the absorbed dose, the ambient dose equivalent, the mean quality factor, and the dose distribution as a function of lineal energy. Data were collected at maximum solar activity in 1991-1992 and at minimum activity in 1996-1998. The lowest mean dose rate measured was 3 microSv h(-1) during a Paris-Buenos Aires flight in 1991. The highest rates were 6.6 microSv h(-1) during a Paris-Tokyo flight on a Siberian route and 9.7 microSv h(-1) on Concorde in 1996-1997. The mean quality factor is around 1.8. The corresponding annual effective dose, based on 700 h of flight for subsonic aircraft and 300 h for Concorde, can be estimated at between 2 mSv for the least-exposed routes and 5 mSv for the more-exposed routes. PMID- 10790274 TI - Role of mast cells in early and delayed radiation injury in rat intestine. AB - Mast cell hyperplasia is a characteristic feature of many inflammatory and fibrotic conditions, including intestinal radiation injury (radiation enteropathy). This study used mast cell-deficient rats to define the role of mast cells in the mechanisms underlying early radiation-induced mucosal injury and delayed intestinal wall fibrosis. Mast cell-deficient (Ws/Ws) mutant rats and mast cell-competent (+/+) littermates were used. A 4-cm loop of ileum was exposed to 21 Gy single-dose radiation. Irradiated and unirradiated intestine were examined at 2 or 26 weeks using quantitative histology and morphometry. Quantitative immunohistochemistry was used to assess transforming growth factor beta (Tgfb), myeloperoxidase, and epithelial and smooth muscle cell proliferation. Collagen content was measured colorimetrically, and steady-state Tgfb1 mRNA was determined with fluorogenic probe RT-PCR. Compared to +/+ rats, Ws/Ws animals exhibited strikingly exacerbated mucosal injury but minimal reactive intestinal wall fibrosis. Ws/Ws rats exhibited less radiation-induced intestinal smooth muscle cell proliferation and collagen accumulation than +/+ littermates. Tgfb expression increased to a similar extent in Ws/Ws and +/+ rats. Unirradiated intestine from Ws/Ws and +/+ rats did not differ significantly. Mast cells protected the intestinal mucosa during the early phase of radiation enteropathy and promoted intestinal fibrosis after the breakdown of the mucosal barrier. Mast cells may be required for Tgfb to exert its full fibrogenic effect in radiation enteropathy. PMID- 10790275 TI - Induction of growth factors in rat cardiac tissue by X irradiation. AB - To investigate the relationship between angiogenic growth factors and endothelial enzyme activity in capillaries after injury of rat cardiomyocytes caused by X irradiation, 7-week-old male Wistar rats were anesthetized with pentobarbitone and their hearts irradiated (X rays, 20 Gy) through a hole in the lead casing in which they were enclosed. The hearts were excised at 1 h, 1 week and 3 weeks after irradiation. Left ventricular cross sections were stained for capillary enzymes by double staining for two endothelial enzymes, alkaline phosphatase (AP) and dipeptidylpeptidase IV (DPP), immunohistochemically stained for basic fibroblast growth factor (Fgf, also known as bFgf) and vascular endothelial growth factor (Vegf), and stained for nick end-labeling of DNA by the TUNEL method. Staining for distribution of AP in the arteriolar portion was reduced at both 1 and 3 weeks after irradiation with 20 Gy, but staining for DPP in the venular portion was unchanged, suggesting a close relationship between growth factors and injury of the arteriolar capillary portion. Fgf and Vegf proteins were present within the cytoplasm of the cardiomyocytes, or around capillaries, 1 h, 1 week and 3 weeks after irradiation. Many TUNEL-stained cardiomyocyte nuclei were observed at 1 h, but they had decreased markedly at 1 week and had almost disappeared by 3 weeks after irradiation. Thus Fgf and Vegf were induced concomitantly with the decrease in the staining for endothelial AP by 20 Gy X irradiation, which also caused microeffects as indicated by TUNEL staining of many nuclei at 1 h postirradiation. PMID- 10790276 TI - Radiation-induced DNA damage in tumors and normal tissues. VI. Estimation of the hypoxic fraction of experimental tumors. AB - For several years, we have concentrated our efforts on validating the use of radiation-induced DNA strand breaks and DNA-protein crosslinks to assess the oxygenation status of tumors and normal tissues. We have demonstrated that (1) the oxygen dependence of strand break formation is identical to that of radiation induced cell killing; (2) the oxygen dependence of DNA-protein crosslink formation is the mirror image of that of radiation-induced cell killing; and (3) the formation of these radiation-induced DNA lesions is predominantly dependent on the oxygen concentration near the DNA and is independent of the cell type, metabolic status, proliferative status, pH of the surrounding environment, and composition or properties of the proteins tightly associated with the DNA. In the present study, the hypoxic fraction of three experimental tumors was estimated using our assay of radiation-induced DNA damage. The average hypoxic fraction of a large number of tumors estimated with this assay of radiation-induced DNA damage for (1) WiDR human colorectal carcinoma xenografts (40.8 +/- 4.2%), (2) 66 mouse mammary adenocarcinoma tumors (41.8 +/- 3.1%), and (3) subcutaneous tumors grown from 9L rat brain tumor cells (95% CI =-8.2-4.2%) was not statistically different from that of a large number of tumors measured for each of these tumor models by the paired survival curve method (38.3 +/- 6. 3%, 28.9 +/- 5.5%, 95% CI = 2.2-4.4%, respectively). When the hypoxic fraction measured by the alkaline elution method on one half of an individual tumor was compared to that measured by the paired survival curve method on the other half of the same tumor, no statistical correlation was found for either 66 or WiDR tumors. Although this assay of radiation-induced DNA damage can be used effectively in the laboratory to answer a number of important questions about the oxygenation status of animal tumors and normal tissues, failure to reliably estimate the hypoxic fraction of individual tumors and technical considerations make it unlikely that the assay can be used in the clinic to estimate the hypoxic fraction of human tumors. PMID- 10790277 TI - The effects of fractionated gamma irradiation on induction of mammary carcinoma in normal and estrogen-treated rats. AB - The effects of dose fractionation on induction of mammary carcinoma were studied in normal and estrogen-treated female rats of the inbred WAG/Rij strain. Groups of 40 animals received total-body doses of 1 or 2 Gy of (137)Cs gamma radiation, administered in fractions of 2.5, 10 or 40 mGy with intervals of 12 h, or in fractions of 10 mGy with intervals of 2, 5 or 24 h. The irradiations were started at the age of 8 weeks. Estrogen treatment was accomplished by implantation of a pellet containing estrogen at the age of 6 weeks. All mammary tumors were resected and classified histologically as carcinoma or fibroadenoma. The age specific incidence of mammary carcinoma was compared with that in control groups of unirradiated normal or estrogen-treated rats and was expressed as excess normalized risk, using lifetime statistical analysis with both parametric and nonparametric methods. The data were also compared to the results of single-dose experiments reported in previous papers. Fractionated irradiation increased the risk of mammary cancer in both normal and estrogen-treated rats compared to the corresponding unirradiated control group. The excess normalized risk per unit of total dose was approximately equal with or without estrogen treatment. Without estrogen treatment, the effects of the single-dose and fractionated irradiations were approximately equal. In estrogen-treated animals, however, single-dose irradiation was up to 15 times more carcinogenic than the fractionated exposures. This fractionation effect appeared to vanish for total doses below approximately 0.3 Gy. With estrogen treatment, the excess normalized risk was significantly higher for dose fractions of 40 mGy than for fractions of 10 mGy. The risk was also markedly higher for fractionation intervals of 2 or 5 h than for intervals of 12 or 24 h. The results of these experiments show that the effects of dose fractionation on the induction of mammary carcinoma may depend on hormonal status, the total dose delivered, the dose per fraction, and the fractionation interval. PMID- 10790278 TI - Detection of individual differences in radiation-induced apoptosis of peripheral blood lymphocytes in normal individuals, ataxia telangiectasia homozygotes and heterozygotes, and breast cancer patients after radiotherapy. AB - Quantification of radiation-induced apoptosis in peripheral blood lymphocytes (PBLs) has been proposed as a possible screening test for cancer-prone individuals and also for the prediction of normal tissue responses after radiotherapy. We have used the TUNEL assay (terminal transferase nick-end labeling) 24 h after irradiation with 4 Gy at high dose rate to assess interindividual differences in radiation-induced apoptosis between (1) a panel of normal individuals, (2) ataxia telangiectasia (AT) homozygotes and heterozygotes, and (3) breast cancer patients who had received radiotherapy 8-13 years ago, including a number of patients who had suffered adverse responses to radiation. With this protocol, we show clear differences in radiation-induced apoptosis between individuals, and good reproducibility in the assay. In agreement with previous reports using EBV-transformed lymphoblasts, we show a very poor induction of apoptosis in AT homozygotes and a reduced level in AT heterozygotes compared to normal individuals. A similar reduced level compared to normal individuals was seen in the breast cancer patients. Despite a wide range of values in the breast cancer patients and good reproducibility on repeat samples, there was no correlation of rates of apoptosis with the severity of breast fibrosis, retraction or telangiectasia. The reduced rate of apoptosis observed in the breast cancer cases may be associated with genetic predisposition to breast cancer; however, we conclude that assays of lymphocyte apoptosis are unlikely to be of use in predicting normal tissue tolerance to radiotherapy. PMID- 10790279 TI - Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes. AB - We have compared dose-rate effects for gamma-radiation-induced apoptosis and micronucleus formation in human lymphocytes. Long-term assessment of individual radiation-induced apoptosis showed little intraindividual variation but significant interindividual variation. The effectiveness of radiation exposure to cause apoptosis or micronucleus formation was reduced by low-dose-rate exposures, but the reduction was apparent at different dose rates for these two end points. Micronucleus formation showed a dose-rate effect when the dose rate was lowered to 0.29 cGy/min, but there was no accompanying cell cycle delay. A further increase in the dose-rate effect was seen at 0.15 cGy/min, but was now accompanied by cell cycle delay. There was no dose-rate effect for the induction of apoptosis until the dose rate was reduced to 0.15 cGy/min, indicating that the mechanisms or signals for processing radiation-induced lesions for these two end points must be different at least in part. There appear to be two mechanisms that contribute to the dose-rate effect for micronucleus formation. One of these does not affect binucleate cell frequency and occurs at dose rates higher than that required to produce a dose-rate effect for apoptosis, and one affects binucleate cell frequency, induced only at the very low dose rate which coincidentally produces a dose-rate effect for apoptosis. Since the dose rate at which cells showed reduced apoptosis as well as a further reduction in micronucleus formation was very low, we conclude that the processing of the radiation-induced lesions that induce apoptosis, and some micronuclei, is very slow in quiescent and PHA stimulated lymphocytes, respectively. PMID- 10790281 TI - Relative biological effectiveness of neutrons: induction of malformations in a mouse strain with genetic predisposition. AB - Female hybrid mice (C57BL/6J x HLG) were exposed to 0.5 Gy of cyclotron neutrons (mean energy 5.8 MeV) after mating with HLG males when embryos were at the early zygote stage. Uterine contents were examined on gestation day 19 for prenatal mortality and malformed development. The results were compared to those obtained after irradiation of zygotes with 1.0 and 1.7 Gy X rays. The number of malformed fetuses was greater than for the unirradiated controls. Gastroschisis was the main observation, but exencephaly was also observed. The neutron relative biological effectiveness (RBE) for the induction of lethal events and malformations in the [(C57BL/6J x HLG)F(1)x HLG] mice was estimated. This value is higher than that for the RBE calculated for the HLG strain, which is known to be more sensitive to radiation than the hybrids. PMID- 10790282 TI - The immunobiology of low-dose total-body irradiation: more questions than answers. AB - Low-dose total-body irradiation (TBI) is used in the treatment of chronic lymphocytic leukemia and low-grade non-Hodgkin's lymphoma. The usual practice is to give very low individual fractions (0.1 to 0.25 Gy) several times a week, to a total dose of 1.5 to 2 Gy. Despite this low dose, low-dose TBI can induce long term remissions in the majority of patients. Immune enhancement, rather than direct radiation cell killing, is one of the suggested mechanisms by which low dose TBI can exert its effect. Data from animal experiments have shown that low dose TBI could enhance the immune response through (1) augmenting the proliferative reactive response of the T cells to mitogenic stimulation; (2) altering cytokine release, particularly the activation of interferon gamma and Il2 production; (3) increasing the expression of Il2 receptors on the T-cell surface; (4) facilitating signal transduction in T lymphocytes; (5) increasing splenic catecholamine content and lowering the serum corticosterone level; and (6) eliminating a particularly radiosensitive subset of the suppressor T cells. Data for humans, though scarce, suggest that at least some of these mechanisms occur in patients treated with low-dose TBI. Whether these immunomodulatory effects are responsible for the clinical outcome is not yet clear. Much is still unknown about the immunobiology of low-dose TBI, its clinical potential, and the possible synergism with chemotherapy, biological response modifiers, or immunotherapy. This lack of comprehensive knowledge hampers the optimal and widespread use of this intriguing and potentially useful treatment modality in clinical practice. PMID- 10790280 TI - Acute effects of whole-body proton irradiation on the immune system of the mouse. AB - The acute effects of proton whole-body irradiation on the distribution and function of leukocyte populations in the spleen and blood were examined and compared to the effects of photons derived from a (60)Co gamma-ray source. Adult female C57BL/6 mice were exposed to a single dose (3 Gy at 0.4 Gy/min) of protons at spread-out Bragg peak (SOBP), protons at the distal entry (E) region, or gamma rays and killed humanely at six different times thereafter. Specific differences were noted in the results, thereby suggesting that the kinetics of the response may be variable. However, the lack of significant differences in most assays at most times suggests that the RBE for both entry and peak regions of the Bragg curve was essentially 1.0 under the conditions of this study. The greatest immunodepression was observed at 4 days postexposure. Flow cytometry and mitogenic stimulation analyses of the spleen and peripheral blood demonstrated that lymphocyte populations differ in radiosensitivity, with B (CD19(+)) cells being most sensitive, T (CD3(+)) cells being moderately sensitive, and natural killer (NK1.1(+)) cells being most resistant. B lymphocytes showed the most rapid recovery. Comparison of the T-lymphocyte subsets showed that CD4(+) T helper/inducer cells were more radiosensitive than the CD8(+) T cytotoxic/suppressor cells. These findings should have an impact on future studies designed to maximize protection of normal tissue during and after proton radiation exposure. PMID- 10790283 TI - The electric and magnetic fields research and public information dissemination (EMF-RAPID) program. AB - In the United States, public concern that exposure to power-line fields was linked to cancer led to the establishment of a Congressionally mandated program, the Electric and Magnetic Fields Research and Public Information Dissemination (EMF-RAPID) Program. A major goal of the program was to "determine whether or not exposures to electric and magnetic fields produced by the generation, transmission, and use of electrical energy affect human health". Between 1994 and 1998, the EMF-RAPID program spent approximately $41 million on biological research. Much of the work funded by the EMF-RAPID program has not yet been published in the peer-reviewed literature. The U.S. National Institute of Environmental Health Sciences (NIEHS) asked that Radiation Research publish this special issue in an attempt to remedy this publication gap. The issue includes reviews of studies that were done to assess the biological plausibility of claims that power-frequency fields caused leukemia and breast cancer. The issue continues with two teratology studies and one immunology study. The section of the issue covering in vitro studies begins with an overview of the efforts NIEHS made to replicate a wide range of reported effects of power-frequency fields and continues with four papers reporting the absence of effects of power-frequency fields on the expression of stress-response genes and oncogenes. Other reports of in vitro studies and studies of mechanisms cover cytotoxicity, gap junction intracellular communication, calcium ion transport across the plasma membrane, and intracellular electric fields. PMID- 10790284 TI - Magnetic fields and mammary cancer in rodents: a critical review and evaluation of published literature. AB - Epidemiological data suggesting a possible increase in breast cancer risk in male electricians have raised concerns about the relationship between exposure to power-frequency magnetic fields and breast cancer. In this paper, we review the results of animal studies that are relevant to identifying possible increases in breast cancer risk resulting from exposure to 50 or 60 Hz magnetic fields. Three large-scale chronic bioassays of carcinogenesis in rats or mice exposed to magnetic fields for 2 years demonstrated no increases in the incidence of mammary cancer; it is generally accepted that power-frequency magnetic fields have little or no activity as a complete carcinogen in the rodent mammary gland. Findings from one laboratory, though inconsistent, suggest that magnetic fields may stimulate mammary neoplasia in rats treated with a chemical carcinogen. However, studies conducted in two other laboratories failed to confirm these findings; rats exposed to magnetic fields demonstrated patterns of tumor incidence, multiplicity, size and latency that were generally similar to those in sham exposed controls. Where differences were seen, the groups exposed to magnetic fields generally had fewer mammary tumors than did sham-exposed controls. On this basis, evaluations of the activity of 50 or 60 Hz magnetic fields in models of multistage mammary cancer in rodents have generally been negative; positive findings have been reported from only one laboratory. The totality of rodent data does not support the hypothesis that power-frequency magnetic-field exposure enhances mammary cancer in rodents, nor does it provide experimental support for possible epidemiological associations between magnetic-field exposure and increased breast cancer risk. PMID- 10790285 TI - Leukemia and lymphoma incidence in rodents exposed to low-frequency magnetic fields. AB - A weak association between residential or occupational exposure to electric and magnetic fields (50/60 Hz fields) and an increased incidence of leukemia has been reported. Numerous animal studies have evaluated the potential association between magnetic-field exposure and leukemia. These include long-term (up to 2(1/2) years) bioassays, initiation/promotion studies, investigations in transgenic models, and tumor growth studies. Exposure to 60 Hz circularly polarized magnetic fields at 1,400 microT for 28 months did not affect lymphoma incidence in mice. The study included over 2000 C57BL/6J mice. In another study, 1000 B6C3F(1) mice exposed to 60 Hz magnetic fields up to 1000 microT for 2 years showed no increase in lymphomas. Approximately 400 transgenic Emu-Pim1 mice exposed to 50 Hz fields up to 1000 microT for up to 18 months had no increased incidence of leukemia. Similarly, Trp53(+/-) mice and Pim1transgenic mice exposed to 60 Hz magnetic fields for 23 weeks showed no increased incidence of lymphoma. Three studies in F344 rats exposed to 50 or 60 Hz magnetic fields up to 5 mT showed no increased incidence of leukemia. The combined animal bioassay results are nearly uniformly negative for magnetic-field exposures enhancing leukemia and weaken the possible epidemiological association between magnetic-field exposures and leukemia in humans as suggested by epidemiological data. PMID- 10790286 TI - Evaluation of the developmental toxicity of 60 Hz magnetic fields and harmonic frequencies in Sprague-Dawley rats. AB - Experimental data suggest that exposure to the 50 and 60 Hz sinusoidal components of power-frequency magnetic fields (MFs) does not have an adverse impact on fetal development. However, the possible developmental toxicity of MF harmonics has not been investigated. This study was designed to determine whether exposure to 180 Hz MFs (third harmonic), alone or in combination with 60 Hz MFs, induces birth defects in Sprague-Dawley rats. Groups of sperm-positive dams (> or =20/group) were exposed for 18.5 h per day from gestation days 6 through 19 to (1) ambient MFs only (<0.0001 mT; sham controls); (2) 60 Hz MFs at 0.2 mT; (3) 180 Hz MFs at 0.2 mT; or (4) 60 Hz + 180 Hz MFs (10% third harmonic; total field strength = 0.2 mT). Litter size, litter weight, percentage live births, sex ratio, and number of resorption sites were determined for each dam, and gross external, visceral, cephalic and skeletal examinations were performed on all fetuses. MF exposure had no significant effects on litter size, litter weight, or fetal development. With the exception of common rib variants, the incidence of fetal anomalies was comparable in all groups. A small increase in the incidence of rib variants was seen in the group exposed to 60 Hz + 180 Hz MFs; however, the incidence of rib variants in this group was similar to that in historical controls from our laboratory. These data extend the existing database on developmental toxicity of MFs by demonstrating that exposure to 180 Hz MFs, either alone or superimposed on an underlying 60 Hz signal, does not induce biologically significant developmental toxicity. These data do not support the hypothesis that exposure to power-frequency MFs is an important risk factor for fetal development. PMID- 10790287 TI - Developmental profiles of growth-associated protein (Gap43), Ngfb, Bndf and Ntf4 mRNA levels in the rat forebrain after exposure to 60 Hz magnetic fields. AB - Fischer 344 rats were exposed to 60 Hz magnetic fields (EMFs) during gestation and lactation. Rats received continuous exposure to 2-, 200- or 1000-microT magnetic fields for 18.5 h per day, 7 days a week, or sham exposure (sham controls). During postnatal development, on postnatal days 1, 3, 6, 9, 15 and 20, forebrain tissue from male pups was examined for alterations in mRNA level for developmentally regulated central nervous system-specific proteins. Alterations in these factors during critical periods of development could result in alterations in the final neural network. Gap43 (growth-associated protein 43) mRNA was measured by Northern hybridization as a developmental indicator of axonal growth during the development of the neuron. Between postnatal days 1 and 9, detectable levels of Gap43 mRNA displayed a similar pattern across all sham control and exposure groups. In addition to Gap43, mRNA levels for the nervous system-specific growth factors ciliary neurotrophic factor (Cntf), brain-derived neurotrophic factor (Bdnf), beta nerve growth factor (Ngfb), neurotrophin-3 (Ntf3), and neurotrophin-4 (Ntf4) were examined by RNase protection assay. While there is public concern for developmental neurotoxicity associated with exposure to EMFs, these data, generated from animals exposed to 2-, 200- or 1000-microT magnetic fields during both gestational and lactational periods of development, suggest that under these conditions no significant alterations in these critical factors for brain development occur. PMID- 10790288 TI - Evaluation of in vitro effects of 50 and 60 Hz magnetic fields in regional EMF exposure facilities. AB - A weak association between magnetic-field exposure and increased incidences of cancer has been reported. While alterations in cellular processes after in vitro magnetic-field exposures have also been reported to provide plausibility for this association, other laboratories have been unable to repeat the findings. As part of an accelerated electric- and magnetic-field (EMF) research program, the National Institute of Environmental Health Sciences with the Department of Energy identified the replication of the published positive effects as a priority. Regional EMF exposure facilities were established to investigate major in vitro effects from the literature. These included effects on gene expression, intracellular calcium, colony growth in soft agar, and ornithine decarboxylase activity. The laboratories that first reported these effects provided experimental protocols, cell lines, and other relevant experiment details. Regional facility studies included sham/sham exposures (no applied field in either chamber) and were done in a blinded fashion to minimize investigator bias. In nearly all experiments, no effects of magnetic-field exposure were found. The effort provided insight into dealing with the difficulty of replication of subtle effects in complex biological systems. Experimental techniques provided some clues for the differences in experimental results between the regional facility and the original investigator. Studies of subtle effects require extraordinary efforts to confirm that the effect can be attributed to the applied exposure. PMID- 10790289 TI - Exposure to low-frequency electromagnetic fields does not alter HSP70 expression or HSF-HSE binding in HL60 cells. AB - Environmental exposure to extremely low-frequency electromagnetic fields (ELF EMFs) has been identified as a possible contributor to increased cancer incidence and other diseases. In vitro studies designed to probe for biological mechanisms that might explain such relationships have included several studies of gene expression. While gene expression studies have focused on MYC, effects of ELF EMFs on the expression of beta-actin, histone H2B, beta-tubulin, SRC, FOS and JUN have also been reported. In addition, some investigators have reported both an induction of HSP70 expression and an increase in HSF-HSE binding in HL60 (human promyelocytic leukemia) cells after exposure to a 60 Hz magnetic field. In this study, HL60 cells were exposed to a weak 60 Hz magnetic field (6.3 or 8.0 microT) or to a positive control heat shock (42 or 44 degrees C). While heat shock led to reproducible induction of HSP70 expression and HSF-HSE binding, no significant effect of exposure to ELF EMFs on either of these end points was observed. PMID- 10790290 TI - Exposure of Daudi cells to low-frequency magnetic fields does not elevate MYC steady-state mRNA levels. AB - The effect of extremely low-frequency electromagnetic field (ELF EMF) exposures to human health has been widely debated. Epidemiological studies have found a possible correlation between increased cancer incidence and environmental ELF EMF exposures. Results from in vitro studies performed to examine the possible underlying bioeffects of ELF EMFs have varied greatly. Reported effects range from robust and reproducible effects to undetectable. In this study, Daudi cells were exposed to 60 Hz magnetic fields for 20, 40 or 60 min at flux densities of 12.5, 50, 100 or 500 microT. Exposures were performed in the Regional ELF-EMF Exposure Facility (Rockville, MD) to minimize variables that might contribute to a false positive effect. Exposures included sham/sham, exposed/sham or sham/exposed, and were performed with blinding with respect to type of exposure. 12-O-Tetradecanoylphorbol-13-acetate (TPA) treatment was used as a positive control. Total cellular RNA was isolated using a single-step technique. Human MYC expression was measured by northern blot hybridization as an indicator of the responsiveness of Daudi cells to experimental conditions. Beta-2-microglobulin (B2M) expression was measured simultaneously as an internal control. Exposure to a 60 Hz magnetic field did not significantly alter MYC expression in Daudi cells under any of the exposure conditions. PMID- 10790291 TI - Expression analysis of human HL60 cells exposed to 60 Hz square- or sine-wave magnetic fields. AB - A total of 960 complementary DNA (cDNA) clones from an HL60 cell cDNA library were screened to discover genes that were differentially expressed in HL60 cells exposed to 60 Hz square-wave magnetic fields (MFs) compared to sham-exposed cells. Square-wave fields are rich in odd harmonic frequency content. We used a two-gel cDNA library screening method (BIGEL) to identify treatment-induced alterations in gene expression. Four cDNA clones were tentatively identified as differentially expressed after exposure to square-wave MFs at 2 mT for 24 h. BIGEL-identified genes (GenBank accession number) corresponding to these clones were: TI227H (D50525), EST Homo sapiens partial cDNA (Z17814), human ribosomal protein S13 (L01124), and AICAR transformylase mRNAs (D82348). The differences in mRNA levels were not confirmed in test compared to experimental cells by Northern analysis. In other experiments, we used concurrent exposure to 60 Hz sine- or square-wave MFs (0 or 2 mT, duration of 3 or 24 h, no postexposure delay). In addition to the four BIGEL genes, we also investigated MYC, HSP70, RAN and SOD1. In the case of MYC and HSP70, square-wave MFs appeared to exhibit more marked alterations when compared to sinusoidal waveforms, but the overall results indicated no effect of possible differential magnetic-field-induced expression of all eight genes. In contrast, alterations of mRNA levels were observed for seven genes after exposure to X irradiation, hyperthermia and TPA. These results are contrary to previously proposed similarities between the action of these agents and MF effects on gene transcription. PMID- 10790293 TI - 60 Hz magnetic fields do not enhance cell killing by genotoxic chemicals in Ataxia telangiectasia and normal lymphoblastoid cells. AB - Ataxia telangiectasia (AT) is an inherited autosomal recessive disease characterized by increased risk of cancer, immune deficiency, and neurodegeneration. Cells cultured from AT patients are highly sensitive to genotoxic agents and are deficient in cell cycle arrest after exposure to ionizing radiation. In consideration of their sensitivity to both ionizing and nonionizing radiation, AT cells may provide a sensitive model system to study the biological activity of other components of the electromagnetic spectrum. To characterize the effects of power-frequency (60 Hz) magnetic fields (MFs) in AT cells, we compared responses of AT and normal lymphoblast cells to sinusoidal MFs at 1.0 mT, either alone or in combination with the genotoxic agents mitomycin C or streptonigrin. The MF alone had no effect on cell growth or survival in a clonogenic assay in either AT or normal cells. The MF also had no effect on induction of cell death by mitomycin C or streptonigrin in either cell type. AT cells do not demonstrate differential sensitivity to MF exposure. These results do not support the hypothesis that MFs interact with genotoxic agents to induce adverse biological effects in either normal or genetically susceptible human cells. PMID- 10790292 TI - Expression of cancer-related genes in human cells exposed to 60 Hz magnetic fields. AB - Exposure to 60 Hz magnetic fields (MFs) may be a risk factor for human cancer. One mechanism through which MFs could influence neoplastic development is through alterations in the expression of cancer-related genes. Previous molecular studies of the action of MFs have measured effects on a limited number of genes. In the present studies, arrays containing cDNAs for 588 cancer-related genes were used to approach the hypothesis that the biological activity of MFs is mediated by alterations in gene expression. Cultures of normal (HME) and transformed (HBL 100) human mammary epithelial cells and human promyelocytic leukemia (HL60) cells were exposed to MFs at field strengths of 0, 0.01 or 1.0 mT for 24 h. Several genes were identified in MF-exposed cells whose expression was increased by at least twofold or decreased by 50% or more. However, no gene was found to be differentially expressed in each of three independent exposures for any cell type, and no relationship between exposure intensity and differential gene expression was found. These studies failed to identify a plausible genetic target for the action of MFs in human cells, and they provide no support for the hypothesis that MF exposure alters the expression of genes that are involved in cancer development. PMID- 10790294 TI - Cellular communication in clone 9 cells exposed to magnetic fields. AB - Magnetic-field exposure (45 Hz B(a.c.) over a flux density range of 7.7 to 49.9 microT r.m.s. with parallel B(d.c.) of 36.6 microT) has been reported by Blackman and coworkers to inhibit gap junction intercellular communication in Clone 9 cells treated with chloral hydrate for 24 h prior to field exposure in accord with predictions of the ion parametric resonance model. The study reported here is an attempt to reproduce this effect. Baseline experiments showed that growth in culture and state of confluence at time of addition of chloral hydrate were comparable in both laboratories. PMA inhibited cell-cell communication in a dose dependent manner, similar to the results of Blackman and coworkers, whereas cells in the present study were somewhat more sensitive to chloral hydrate than reported by Blackman and coworkers. A total of 38 exposure experiments were undertaken using a 45 Hz magnetic field with a flux density of 23.8 microT r.m.s., in parallel with a 36.6-microT static magnetic field for 40 to 45 min, after pretreatment with 2.5 mM chloral hydrate for 24 h. In 14 unblinded experiments, a small but statistically significant effect of magnetic-field exposure was observed, but due to the subjective nature of the assay, it was deemed essential to carry out blinded experiments. The remaining 24 experiments were blinded. In 15 blinded experiments, cells purchased from the American Type Culture Collection and grown only in this laboratory were used, while in 9 experiments, the cells had originally been grown in Blackman's laboratory and were subsequently sent to this laboratory. There was no statistically significant effect of magnetic-field exposure on gap junction intercellular communication in these blinded experiments using either cell line. PMID- 10790295 TI - Power-frequency electromagnetic fields and the capacitative calcium entry system in SV40-transformed Swiss 3T3 cells. AB - The present study was designed to test the hypothesis that a 60 Hz electromagnetic field could affect the influx of calcium ions across the plasma membrane through the so-called capacitative calcium entry system. Recordings of cytosolic calcium-ion concentrations in SV40-transformed Swiss 3T3 cells were obtained in real time during exposure to magnetic fields ranging from 0.3-50 mT or to sham conditions using the calcium-sensitive photoprotein aequorin. This was done for cell populations whose capacitative entry system was activated by either bradykinin or thapsigargin under a variety of experimental conditions. No effects of the magnetic field were observed on bradykinin-induced calcium transients and, with the exception of a small but statistically significant increase observed in experiments performed at 50 mT, no effects of the fields were observed on baseline calcium levels prior to or after such transients. The magnetic fields also had no effects on the size or kinetics of any of the thapsigargin-induced calcium transients. Overall, the data fail to support the hypothesis tested in this work. PMID- 10790296 TI - Suppression of a differentiation response in MC-3T3-E1 osteoblast-like cells by sustained, low-level, 30 Hz magnetic-field exposure. AB - Extremely low-frequency (ELF) magnetic fields have been reported to be capable of influencing both tissue remodeling and cell phenotypic expression in culture. However, whether the cells or tissues respond directly to the magnetic flux or to the electric field induced by the time-changing magnetic flux remains a controversial topic. To address this question, we developed an osteoblast cell assay based on the activity of alkaline phosphatase, an enzyme whose activity is up-regulated during the differentiation of bone cells. MC-3T3-E1 cells plated at a confluent density were allowed to proceed through the differentiation process for 3 days, after which they were exposed to a 30 Hz, 1.8-mT r.m.s. magnetic field inducing a spatially varying electric field with a maximum intensity of 0.9 mV/m r.m.s. In situ assays of alkaline phosphatase activity at 4, 8, 16 and 64 h demonstrated a progressive inhibition of enzyme activity, the pattern of which maps to the intensity of the induced electric field (R(2) = 0.5, P<0.001). We interpret these results to indicate that cells are capable of responding to ELF induced electric fields at intensities below 1 mV/m, and that the principal effect on cells is an inhibition of differentiation. PMID- 10790297 TI - Calculated electric field induced in bundles of long cells in the human body when it is exposed to low-frequency electric fields. AB - A theoretical analysis is made of the electric field induced in the interior of long cylindrical cells arranged in bundles of parallel cells embedded in the saline fluid in the human body exposed to extremely low-frequency (ELF) and very low-frequency (VLF) electric fields parallel to the length of the cells. Such bundles characterize muscle. The analysis parallels in many respects the earlier study of isolated long cells (King and Wu, Phys. Rev. E 58, 2363-2369, 1998) but has important added complications. It leads to a similar result: The field inside the membrane of each long cell is the same as the field just outside the membrane in the saline fluid; i.e., the membrane provides no shielding. While one might expect the presence of the neighboring cells to provide a shielding effect for each cell, the results of this study show quantitatively that this is not the case. In fact, the electric field in the interior of each of a bundle of long cells is seen to increase very slightly (of the order of 0.4%) when the cells in the bundle are moved close enough to be practically in contact. PMID- 10790298 TI - Modulation of natural killer cell function after exposure to 60 Hz magnetic fields: confirmation of the effect in mature B6C3F1 mice. AB - In a previous study, we demonstrated that subchronic exposure to pure, linearly polarized 60 Hz magnetic fields (MFs) at flux densities ranging from 0.002 to 1.0 mT induced a modest but statistically significant and reproducible suppression of NK cell activity in young adult B6C3F(1) mice. NK cell activity in mice declines with age and is known to be suboptimal in older animals. The present study was designed to determine if the same MF exposure regimens will suppress NK cell activity in mature (i.e. more than 1 year old) animals. Extending our previous findings, a modest suppressive effect of MFs on NK cell activity in B6C3F(1) mice was found when subchronic exposure was initiated in animals held in quarantine for 1 year prior to exposure. These data demonstrate that MF exposure suppresses NK cell activity in both young and mature adult B6C3F(1) mice. However, because chronic exposure to the same MF parameters used in the NK function studies does not increase the incidence of neoplasia in B6C3F(1) mice, this statistically significant inhibition of NK cell function appears to be of limited biological significance. PMID- 10790299 TI - Cell viability and growth in a battery of human breast cancer cell lines exposed to 60 Hz magnetic fields. AB - Epidemiological data suggest that exposure to power-frequency (50/60 Hz) magnetic fields (MFs) may be a risk factor for breast cancer in humans. To determine whether MFs affect human breast cancer cells, we measured viability, growth and cytotoxicity in a battery of breast cancer cell lines after in vitro MF and sham exposure. Cells of three estrogen receptor-positive human breast cancer cell lines (MCF-7, ZR-75-1 and T-47D) and one estrogen receptor-negative human breast cancer cell line (MDA-MB-231) and normal (nontransformed) human breast epithelial cells were exposed to MFs (1 mT) or sham fields (<0.0001 mT) for 72 h. Cell viability was determined using the sulforhodamine B (SRB) assay at 0 and 72 h after the MF exposure period. Cell growth was measured as the change in SRB dye uptake over 72 h after MF exposure. MF exposure had no effect on cell viability or growth in any cell type examined. Similarly, MF exposure had no effect on cytotoxicity induced by exposure to the retinoid N-(4-hydroxyphenyl)retinamide. These data do not support the hypothesis that MF exposure stimulates growth of breast cancer cells. PMID- 10790300 TI - Selenium determination in biological fluids using Zeeman background correction electrothermal atomic absorption spectrometry. AB - Procedures for the direct determination of total selenium in urine, serum, and blood using electrothermal atomic absorption spectrometry are presented. In the selected experimental conditions, Zeeman correction is mandatory to compensate for the high background signals. The sample diluted and containing 0.1% (w/v) Triton X-100 is introduced directly into the electrothermal atomizer. A solution containing 15% (w/v) hydrogen peroxide, 0.65% (w/v) nitric acid, and 0.5% (w/v) nickel is injected separately into the atomizer. Calibration is carried out using the standard additions method. The detection limit is 30 pg selenium. If palladium, instead of nickel, is used as the chemical modifier, calibration can be carried out against aqueous standards, and the detection limit is 45 pg. In this case, three separate injections are required to prevent precipitation problems in the automatic injector. The reliability of the procedures is checked by analyzing three certified reference materials and by recovery studies. Mean recoveries are 99.7% for serum, 99.4% for urine, and 100.8% for blood samples. Relative standard deviation values are +/-4.0% for serum, +/-3.9% for urine, and +/-4.5% for blood. PMID- 10790301 TI - Synthesis of peptidyl methylcoumarin esters as substrates and active-site titrants for the prohormone processing proteases Kex2 and PC2. AB - Peptidyl methylcoumarin amides are well established as model substrates for understanding protease specificity, but the corresponding methylcoumarin esters have attracted scant attention despite their potential utility in active-site titration and mechanistic characterization. We have devised techniques for the synthesis and deprotection of extended peptidyl methylcoumarin esters in good to moderate yields, and we have demonstrated their suitability for steady-state characterization and active-site titration of the Saccharomyces cerevisiae processing protease Kex2. Additionally, we have used one of these compounds to active-site titrate the homologous enzyme PC2, which had not previously been feasible using other types of substrates. These compounds should thus prove widely suitable for use as substrates and active-site titrants not only for proteases of the prohormone processing family but also for a wide range of other serine proteases. PMID- 10790302 TI - Estimation of receptor-ligand interactions by the use of a two-marker system in affinity capillary electrophoresis. AB - The study of receptor-ligand interactions by affinity capillary electrophoresis (ACE) requires an accurate form of analysis. Here, we examine the use of two noninteracting standards (markers) in the analysis of binding constant data in ACE studies. This concept is demonstrated using two model systems: carbonic anhydrase B (CAB, EC 4.2.1.1) and arylsulfonamides, and vancomycin (Van) from Streptomyces orientalis and the dipeptide N-acetyl-d-Ala-d-Ala. In this procedure a plug of receptor and noninteracting standards is injected, and analysis of the change in the relative migration time ratio of the receptor, relative to the noninteracting standards, as a function of the concentration of the ligand yields a value for the binding constant. The findings described here demonstrate that data from ACE studies can best be analyzed using two noninteracting standards, yielding values comparable to those estimated using other binding and ACE techniques. PMID- 10790303 TI - Use of bidirectional blots in differential display analysis. AB - We have used bidirectional transfer methods in concert with SMART total cDNA complex probes to sequentially screen differential display arrays. In this report we show the utility of this methodology in examining a manganese superoxide dismutase cDNA fragment which we detected while evaluating the effects of the proinflammatory cytokines IL1-beta, TNF-alpha, and IL6 on human umbilical vein endothelial cell (HUVEC) gene expression. By using parallel hybridization of the bidirectional blots with SMART total cDNA (32)P probes derived from untreated or cytokine-treated HUVECs, differential expression between cell treatments can be clearly evaluated. Subsequent screening using this bidirectional blot method results in detection of modulated cDNA clones. Northern and total cDNA blot hybridization with the cDNA clonal fragment confirmed both modulated expression and the efficacy of this screening method. These procedures allow one to use bidirectional blots to evaluate band modulation on agarose gels which are initially run to evaluate the reamplification of display fragments or to confirm cloned cDNA fragments. Thus, bidirectional blot analysis using SMART total cDNA probes allows direct evaluation of differential display bands from the initial reamplification through plasmid insert cloning, increasing the investigator's ability to eliminate false-positive bands during each step of analysis. PMID- 10790304 TI - Amperometric glucose sensor based on coimmobilization of glucose oxidase and Poly(p-phenylenediamine) at a platinum microdisk electrode. AB - A miniaturized glucose biosensor in which glucose oxidase (GOD) and poly(p phenylenediamine) (poly-PPD) were coimmobilized at the surface of a platinum microdisk electrode was developed and used successfully for amperometric determination of glucose. The performance of sensors prepared at different monomer concentrations and polymerization potentials with different media was investigated in detail. It was found that similarly to poly(o-phenylenediamine) (poly-OPD), (poly-PPD) noticeably eliminated the electrochemical interference of ascorbic acid, uric acid, and l-cysteine. The amperometric response of glucose with the biosensor under optimal conditions exhibited a linear relationship in the range of 5.0 x 10(-5) to 3.0 x 10(-3) M with correlation coefficient 0.9995. According to the Michaelis-Menten equation, the apparent Michaelis constant for glucose and the maximum steady-state current density of the poly-PPD/GOD-modified microelectrode were 3.94 mM and 607.5 microA cm(-2), respectively. The current density of the sensor responding to glucose in the linear range can reach 160 microA cm(-2) mM(-1), which is far greater than that obtained using poly-OPD and poly(phenol) film. In addition, the stability of the sensor was examined over a 2 month period. PMID- 10790305 TI - Fluorescence lifetime microscopy of the sodium indicator sodium-binding benzofuran isophthalate in HeLa cells. AB - The behavior of the sodium indicator sodium-binding benzofuran isophthalate (SBFI) is investigated in HeLa cells by time-resolved fluorescence microscopy. The fluorescence relaxation of SBFI in HeLa cells can be described by a triexponential for intracellular sodium concentration ([Na(+)](i)) between 0 and 90 mM. Changes in [Na(+)](i) affect neither the fluorescence relaxation times (0.21, 0. 60, and 2.7 ns) nor the average decay time (2.2 ns). The preexponential factor of the shortest decay time is negative. However, the ratio of the fluorescence excitation signal at 340 nm to that at 380 nm increases with [Na(+)](i). To elucidate the behavior of SBFI in cells, experiments are performed on SBFI in buffer at various concentrations of sodium, potassium, and bovine serum albumin (BSA) and at various viscosities. The fluorescence decay is triexponential only in the presence of BSA. The relaxation times are independent of [Na(+)] and [BSA]. The preexponential factor of the shortest decay time is negative from a certain [BSA] on, which depends on [Na(+)]. The data indicate that interactions with intracellular components rather than microviscosity influence the SBFI behavior in cells. A model is suggested in which the fluorescence intensities are mainly determined by the signals from the Na(+) subsetSBFI and SBFI subsetprotein complexes. PMID- 10790306 TI - Measurement of phenyllactate, phenylacetate, and phenylpyruvate by negative ion chemical ionization-gas chromatography/mass spectrometry in brain of mouse genetic models of phenylketonuria and non-phenylketonuria hyperphenylalaninemia. AB - Phenylketonuria (PKU) (OMIM 261600) is the first Mendelian disease to have an identified chemical cause of impaired cognitive development. The disease is accompanied by hyperphenylalaninemia (HPA) and elevated levels of phenylalanine metabolites (phenylacetate (PAA), phenyllactate (PLA), and phenylpyruvate (PPA)) in body fluids. Here we describe a method to determine the concentrations of PAA, PPA, and PLA in the brain of normal and mutant orthologous mice, the latter being models of human PKU and non-PKU HPA. Stable isotope dilution techniques are employed with the use of [(2)H(5)]-phenylacetic acid and [2,3, 3-(2)H(3)]-3 phenyllactic acid as internal standards. Negative ion chemical ionization (NICI) GC/MS analyses are performed on the pentafluorobenzyl ester derivatives formed in situ in brain homogenates. Unstable PPA in the homogenate is reduced by NaB(2)H(4) to stable PLA, which is labeled with a single deuterium and discriminated from endogenous PLA in the mass spectrometer on that basis. The method demonstrates that these metabolites are easily measured in normal mouse brain and are elevated moderately in HPA mice and greatly in PKU mice. However, their concentrations are not sufficient in PKU to be "toxic"; phenylalanine itself remains the chemical candidate causing impaired cognitive development. PMID- 10790307 TI - An enzyme-linked lectin assay for alpha1,3-galactosyltransferase. AB - UDP-Gal:Galbeta1-4GlcNAc alpha1,3-galactosyltransferase (alpha3GalT) is responsible for the synthesis of carbohydrate xenoantigen Galalpha1-3Galbeta1 4GlcNAc. In this work a convenient and sensitive assay system for quantification of alpha3GalT activity by enzyme-linked lectin assay (ELLA) with colorimetric detection is described. Microtiter plate wells whose surface had been coated with the polyacrylamide conjugate of the disaccharide Galbeta1-4GlcNAc (acceptor) are incubated with alpha3GalT in the presence of "cold" UDP-Gal as glycosyl donor. Formation of product by enzymatic extension of the glycan chain is detected by the biotinylated plant lectin Viscum album agglutinin. The standard curve for correct quantification of alpha3GalT activity is completed after running standard assays with no (background) or known quantities of enzyme activity. Product formation detected in this manner is proportional to enzyme activity and the concentrations of the acceptor and the glycosyl-donor UDP-Gal. In accordance with the known specificity of alpha3GalT, no enzymatic conversion of Le(x) into GalalphaLe(x) was observed using this assay. Human alphaGal antibodies were isolated using a disaccharide-exposing affinity adsorbent and their specificity was studied. Relative to the application of these natural immunoglobulins as product-detecting tool, the ELLA proved to be more sensitive. PMID- 10790308 TI - Quantitation of arachidonic acid metabolites in small tissue biopsies by reversed phase high-performance liquid chromatography. AB - Arachidonic acid metabolites exert a variety of distinct biological effects on the initiation and resolution of inflammatory diseases and their measurements in tissue can be critical to evaluate their regulatory function during the course of inflammation and to supplement in vitro experiments. The aim of this study was the detection and quantitative analysis of four arachidonic acid metabolites in small-sized biopsies of human periodontal tissues. The biopsies were homogenized and injected directly into a single analytical column of a RP-HPLC system. Detection was performed by a photodiode array detector. Calibration was established by dilutions of authentic standards of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), 12(R)-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), and 15(S)-hydroxy-5,8,11,13-eicosatetraenoic acid (15-HETE). A total of 38 specimens weighing between 19 and 191 mg (wet tissue) were analyzed (mean = 59.9 +/- 30.2 mg). The detection limits were 1 pg for LTB4 and 12-HETE, 0.5 pg for 15 HETE, and 10 ng for PGE2. The concentrations of PGE2 and LTB4 were significantly higher in inflamed than in healthy periodontal tissues (P = 0.0079; P = 0. 0114). 12-HETE was detected in one biopsy (30 pg/g); 15-HETE was not detected. This method of homogenization, extraction, and analysis of arachidonic acid metabolites by RP-HPLC appears to be well suited for studies of human oral biopsies. Only small tissue samples and minimal laboratory equipment were required for a sensitive analysis. PMID- 10790309 TI - Measuring dissociation constants of RNA and aminoglycoside antibiotics by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) has been used to determine the dissociation constants (K(D)s) and binding stoichiometry for tobramycin and paromomycin with a 27-nucleotide RNA construct representing the A-site of the 16S ribosomal RNA. K(D) values determined by holding the ligand concentration fixed are compared with K(D) values derived by holding the RNA target concentration fixed. Additionally, the effect of solution conditions such as the amount of organic solvent present and the amount of salt present in the solution on the K(D) measurement is investigated. It is shown that the preferred method for determining dissociation constants using ESI-MS is holding the RNA target concentration fixed below the expected K(D) and titrating the ligand. K(D) measurements should also be carried out at as high as possible salt concentration to minimize nonspecific binding due primarily to electrostatic interactions. For tobramycin, two nonequivalent binding sites were found with K(D1) = 352 nM and K(D2) = 9 microM. For paromomycin, there is only one binding site with K(D) = 52 nM. PMID- 10790310 TI - Red laser-induced fluorescence energy transfer in an immunosystem. AB - We describe two near-infrared fluorescent squaraine dyes (Sq635 and Sq660), their spectra, their covalent linkage to proteins, and their use as donor and acceptor, respectively, in a fluorescence resonance energy transfer (FRET) immunoassay based on the use of red lasers. The dyes show quantum yields of around 10% in the free form and up to 68% when bound to proteins. If converted into their N hydroxysuccinimide esters, they can be linked to free amino groups of proteins. To improve water solubility, two sulfo groups were introduced. The emission spectrum of Sq635 overlaps the absorption spectrum of Sq660, a fact that makes them a useful pair of dyes for use in FRET immunoassay which is demonstrated for human serum albumin/anti-human serum albumin. PMID- 10790311 TI - Determination of 3-nitrotyrosine by high-pressure liquid chromatography with a dual-mode electrochemical detector. AB - 3-Nitrotyrosine, a product of tyrosine nitration, is useful as a marker for the generation of reactive nitrogen oxide species with short half-lives such as peroxynitrite. A reverse-phase high-pressure liquid chromatographic method using a dual-mode electrochemical detector in series with a photodiode array detector has been developed to determine the levels of 3-nitrotyrosine in biological samples. The principle of this method involves reduction of 3-nitrotyrosine at an upstream gold amalgam electrode and oxidation of the resulting product(s) at a downstream glassy carbon electrode. 3-Nitrotyrosine is quantified by the amount of the current generated at the downstream electrode, and a femtomole detection level can be achieved. The disappearance of the corresponding peak when the electrochemical detector is used only in the single oxidative mode provides additional evidence for the identity of 3-nitrotyrosine in the sample. Tyrosine from the same sample is determined by its UV absorption at 280 nm, thus eliminating the need for an internal standard. With this method a dose-dependent increase of 3- to 10-fold in the levels of protein 3-nitrotyrosine was observed in the blood plasma, and a 2- to 4-fold increase in the lung cytosols, of rats treated with the lung carcinogen and nitrating agent tetranitromethane. PMID- 10790312 TI - An assay for angiotensin-converting enzyme using capillary zone electrophoresis. AB - A sensitive and rapid method was developed for angiotensin-converting enzyme (ACE) activity determination by capillary zone electrophoresis. Hippuryl-l histidyl-l-leucine, a synthetic tripeptide, was used as the ACE-specific substrate. Capillary zone electrophoresis was employed to separate the products of the enzymatic reaction and the ACE activity was determined by quantification of hippuric acid, a result of the enzymatic reaction on the tripeptide. The capillary electrophoresis was performed in a 27 cm x 75 micrometer i.d. fused silica capillary using 200 mM boric acid-borate buffer (pH 9.0) as a run buffer with an applied voltage of 8.1 kV at a capillary temperature of 23 degrees C. The electrophoresis was monitored at 228 nm. Each electrophoretic run requires only a nanoliter of the enzymatic reactant solution, at only 6 min, rendering a powerful tool for the ACE assay. PMID- 10790313 TI - Development of a sensitive chemiluminescent neuraminidase assay for the determination of influenza virus susceptibility to zanamivir. AB - Determination of the sensitivity of influenza viruses to neuraminidase (NA) inhibitors is presently based on assays of NA function because, unlike available cell culture methods, the results of such assays are predictive of susceptibility in vivo. At present the most widely used substrate in assays of NA function is the fluorogenic reagent 2'-O-(4-methylumbelliferyl)-N-acetylneuraminic acid (MUN). A rapid assay with improved sensitivity is required because a proportion of clinical isolates has insufficient NA to be detectable in the current fluorogenic assay, and because some mutations associated with resistance to NA inhibitors reduce the activity of the enzyme. A chemiluminescence-based assay of NA activity has been developed that uses a 1,2-dioxetane derivative of sialic acid (NA-STAR) as the substrate. When compared with the fluorogenic assay, use of the NA-STAR substrate results in a 67-fold reduction in the limit of detection of the NA assay, from 200 pM (11 fmol) NA to 3 pM (0.16 fmol) NA. A panel of isolates from phase 2 clinical studies of zanamivir, which were undetectable in the fluorogenic assay, was tested for activity using the NA-STAR substrate. Of these 12 isolates with undetectable NA activity, 10 (83%) were found to have detectable NA activity using the NA-STAR substrate. A comparison of sensitivity to zanamivir of a panel of influenza A and B viruses using the two NA assay methods has been performed. IC(50) values for zanamivir using the NA-STAR were in the range 1.0-7.5 nM and those for the fluorogenic assay in the range 1. 0-5.7 nM (n = 6). The NA-STAR assay is a highly sensitive, rapid assay of influenza virus NA activity that is applicable to monitoring the susceptibility of influenza virus clinical isolates to NA inhibitors. PMID- 10790314 TI - Biotinylated phosphatidylinositol 3,4,5-trisphosphate as affinity ligand. AB - Phosphatidylinositol 3,4,5-trisphosphate (PIP(3)), a primary output signal of phosphoinositide (PI) 3-kinase, plays a crucial role in diverse cellular processes. Evidence indicates that PIP(3) exerts downstream signaling, in part, by recruiting effector proteins to plasma membranes. Consequently, identification of signaling enzymes with PIP(3)-binding motifs represents a viable approach to understand the mechanism by which specificity of the PI 3-kinase-mediated signaling network is maintained. To address this issue, we have developed biotinylated derivatives of PIP(3) as affinity probes for the purification and characterization of PIP(3)-binding proteins. Considering the relaxed requirement for the acyl moiety in PIP(3) recognition, these biotinylated PIP(3) analogues display two structural features. First, they contain short acyl side chains (C(4) and C(8)), allowing them to be soluble in aqueous milieu. This desirable feature avoids the formation of lipid aggregates, which minimizes nonspecific hydrophobic interactions with proteins. Second, the appended biotin is located at the terminus of the sn-1 acyl side chain, thereby maintaining the integrity of the phosphoinositol head group essential for selective recognition. The utility of these affinity ligands is validated by the purification of recombinant PIP(3) binding proteins, expressed as GST fusion proteins, to homogeneity from bacterial lysates. These include the C-terminal SH2 domain of the p85 subunit of PI 3 kinase and the N-terminal PH domain of PLCgamma1. The efficiency of biotinylated PIP(3) analogues in the purification of these recombinant proteins was approximately 20% of that of glutathione beads PMID- 10790315 TI - A continuous, nonradioactive assay for histone acetyltransferases. AB - Histone acetyltransferases (HATs) catalyze the acetyl-group transfer from acetyl CoA to the epsilon-amino group of specific lysine residues within core histone proteins. HATs and other chromatin-remodeling enzymes have been recently shown to regulate gene activation within specific loci. To facilitate mechanistic studies, we have developed two continuous, nonradioactive assays for the prototypical GCN5 HAT. The CoASH generated in the HAT reactions was continuously measured by using a coupled enzyme system with either alpha-ketoglutarate dehydrogenase or pyruvate dehydrogenase. The CoASH-dependent oxidation of alpha-ketoglutarate or pyruvate is accompanied by the reduction of NAD to NADH, which was measured spectrophotometrically at 340 nm. The steady-state rate constants with substrates acetyl-CoA and a synthetic peptide (corresponding to the first 20 amino acids of H3 histone) were determined. The resulting rate constants were not significantly different between the two coupled assays, providing strong validation of these methods. Rate constants were also determined using the commonly employed radioactive filter-binding assay and compared. The 1.5- to 5-fold lower values obtained in the radioactive end-point assay are discussed in terms of the technical problems and limitations of this assay. The coupled assays should be widely applicable since the production of CoASH is common to all HAT enzymes, regardless of protein substrate. PMID- 10790316 TI - Assay for identification of inhibitors for bacterial MraY translocase or MurG transferase. AB - Bacterial peptidoglycan synthesis is a well-characterized system for targeting new antimicrobial drugs. Formation of the peptidoglycan precursors Lipid I and Lipid II is catalyzed by the gene products of mraY and murG, which are involved in the first and second steps of the lipid cycle reactions, respectively. Here we describe the development of an assay specific for identifying inhibitors of MraY or MurG, based on the detection of radiolabeled [(14)C]GlcNAc incorporated into Lipid II. Assay specificity is achieved with the biotin tagging of the Lipid I precursor UDP-MurNAc-pentapeptide. This allows for the separation and identification of lipid products produced by the enzymatic activity of the MraY and MurG proteins, and thus identification of specific inhibitors. PMID- 10790317 TI - Development of an Aspergillus fumigatus cell-free protein synthesis assay. PMID- 10790318 TI - Detection of DNA using a visible dye, Nile blue, in electrophoresed gels. PMID- 10790319 TI - The effect of cycle number and target size on polymerase chain reaction amplification of polymorphic repetitive sequences. PMID- 10790320 TI - Refractive index of proteins in aqueous sodium chloride. PMID- 10790321 TI - An enzymatic fluorometric assay for fructose. PMID- 10790322 TI - Toward neuroanatomical models of analogy: a positron emission tomography study of analogical mapping. AB - Several brain regions associated with analogical mapping were identified using (15)O-positron emission tomography with 12 normal, high intelligence adults. Each trial presented during scanning consisted of a source picture of colored geometric shapes, a brief delay, and a target picture of colored geometric shapes. Analogous pictures did not share similar geometric shapes but did share the same system of abstract visuospatial relations. Participants judged whether each source-target pairing was analogous (analogy condition) or identical (literal condition). The results of the analogy-literal comparison showed activation in the dorsomedial frontal cortex and in the left hemisphere; the inferior, middle, and medial frontal cortices; the parietal cortex; and the superior occipital cortex. Based on these results as well as evidence from relevant cognitive neuroscience studies of reasoning and of executive working memory, we hypothesize that analogical mapping is mediated by the left prefrontal and inferior parietal cortices. PMID- 10790323 TI - Organization and components of psychomotor ability. AB - We investigated the organization and components of psychomotor abilities by administering a diverse set of cognitive and psychomotor tasks to a group of recent high school graduates (N = 161). Confirmatory factor analyses identified two psychomotor factors: a general factor associated with all psychomotor tests and an orthogonal psychomotor learning factor associated exclusively with practiced psychomotor tests. Path analyses suggested that the general psychomotor factor could be largely accounted for by two cognitive factors, general working memory capacity (r = .67) and an orthogonal time estimation factor (r = .32). Most of the psychomotor learning factor variance was unique, but psychomotor learning was somewhat related to processing speed (r = .49). We conclude that initial psychomotor performance is constrained by working-memory limits and the ability to keep track of time. Practiced psychomotor skill is additionally limited by processing speed, consistent with the literature on the development of process automaticity. The Discussion addresses (a) the small dimensionality of the psychomotor abilities space, (b) our discovery of the importance of time estimation and other cognitive factors in psychomotor learning, and (c) the changing nature of psychomotor skill with practice. PMID- 10790324 TI - On the reliability of implicit and explicit memory measures. AB - Functional dissociations between implicit and explicit memory tests often take the form of large differences between groups or experimental conditions (e.g., amnesics and controls, elderly and younger persons, or persons learning with and without a distracting secondary task) when performance is assessed using explicit memory tests, whereas no difference is observed with implicit memory tests. We argue that the interpretation of such dissociations in terms of the memory processes or systems involved in performance is problematic because the same data pattern would emerge as a result of a mere methodological artifact, that is, the situation that implicit memory tests have low reliability whereas explicit memory tests are fairly reliable measurement instruments. We present reasons for such a reliability difference, and we demonstrate it empirically in Experiments 1a, 1b, and 2. However, our analysis also shows, and Experiment 3 confirms empirically, that implicit memory tests need not necessarily be less reliable measurement instruments than explicit memory tests. PMID- 10790325 TI - Regeneration in vertebrates. AB - One way or another, all species possess the ability to regenerate damaged tissues. The degree of regeneration, however, varies considerably among tissues within a body and among species, with urodeles being the most spectacular. Such differences in regenerative capacity are indicative of specific mechanisms that control the different types of regeneration. In this review the different types of regeneration in vertebrates and their basic characteristics are presented. The major cellular events, such as dedifferentiation and transdifferentiation, which allow complex organ and body part regeneration, are discussed and common molecular mechanisms are pinpointed. PMID- 10790327 TI - CHE-3, a cytosolic dynein heavy chain, is required for sensory cilia structure and function in Caenorhabditis elegans. AB - Forward genetic screens using novel assays of nematode chemotaxis to soluble compounds identified three independent transposon-insertion mutations in the gene encoding the Caenorhabditis elegans dynein heavy chain (DHC) 1b isoform. These disruptions were mapped and cloned using a newly developed PCR-based transposon display. The mutations were demonstrated to be allelic to the che-3 genetic locus. This isoform of dynein shows temporally and spatially restricted expression in ciliated sensory neurons, and mutants show progressive developmental defects of the chemosensory cilia. These results are consistent with a role for this motor protein in the process of intraflagellar transport; DHC 1b acts in concert with a number of other proteins to establish and maintain the structural integrity of the ciliated sensory endings in C. elegans. PMID- 10790326 TI - Participation of a K(+) channel modulated directly by cGMP in the speract-induced signaling cascade of strongylocentrotus purpuratus sea urchin sperm. AB - Speract, a decapeptide from Strongylocentrotus purpuratus sea urchin eggs, transiently stimulates a membrane guanylyl cyclase and activates a K(+)-selective channel that hyperpolarizes sperm. However, previous studies of sperm and of sperm membrane vesicles reached conflicting conclusions about the mechanisms that open these channels. We find that speract hyperpolarizes and increases the cGMP content of flagellar vesicles. We confirm previous findings that intravesicular GTPgammaS and GTP enhance this hyperpolarization, but not GDPbetaS. The G protein activators AlF(-)(4) and mastoparan also are ineffective. Thus, it is unlikely that a G protein participates in the speract response. In contrast, hyperpolarization responses to speract are increased by 3-isobutyl-1 methylxanthine, which preferentially inhibits cGMP-selective phosphodiesterases of sperm, and the 8Br-cGMP derivative hyperpolarizes vesicles in the absence of speract. The responses to speract and to 8Br-cGMP have similar ionic selectivities (K(+) > Rb(+) > > Li(+) > Na(+)) and sensitivities to the channel blockers 4-aminopiridine and 3, 4-dichlorobenzamil, indicating that they likely result from opening of the same K(+) channel. Inhibitors that preferentially inhibit cAMP-selective phosphodiesterases do not alter responses to speract, and permeant cAMP analogs do not hyperpolarize vesicles. In addition, inhibitors of protein kinases and phosphatases fail to alter vesicle hyperpolarization by speract. The increase in vesicular cGMP content produced by speract therefore may directly mediate opening of the channel that hyperpolarizes sperm membrane vesicles. Similar mechanisms presumably operate in intact sperm. PMID- 10790328 TI - An essential role of the neuronal cell adhesion molecule contactin in development of the Xenopus primary sensory system. AB - Contactin is a glycosylphosphatidylinositol-anchored immunoglobulin-like neuronal cell adhesion molecule that has been implicated in cellular interaction during development of the vertebrate central nervous system. Here we report evidence for an essential role of contactin in development of the Xenopus nervous system. Contactin mRNA is detectable by in situ hybridization in subsets of neurons in the brain, primary sensory neurons in the spinal cord, and cells along the trigeminal nerves of tailbud embryos. Contactin immunoreactivities preferentially distribute on axon tracts of the brain, the spinal cord, and the trigeminal sensory nerves. Most prominently, cell bodies and peripheral and spinal axons of primary sensory neurons, Rohon-Beard (RB) cells, are strongly contactin positive. Injection of the contactin overexpression vector into one blastomere of two-cell stage embryos leads to misdirected elongation of the peripheral axons of RB neurons in the injected half. Overexpression of antisense transcript causes depletion of contactin mRNA accumulation and abnormal development of RB neurons. In 52.3% of the injected embryos, RB neurons decrease in number and their peripheral axons in dorsal lateral tracts are defasciculated. These results demonstrate that contactin plays an essential role in development of the Xenopus primary sensory system. PMID- 10790329 TI - More than 95% reversal of left-right axis induced by right-sided hypodermic microinjection of activin into Xenopus neurula embryos. AB - In recent years, genes that show left-right (L-R) asymmetric expression patterns have been identified one after another in vertebrate gastrula-neurula embryos. However, we still have little information about when the irreversible L-R specification is established in vertebrate embryos. In this report, we show that almost 100% of the embryos develop to be L-R-inverted larvae after microinjection of activin molecules into the right lateral hypodermic space of Xenopus neurula embryos. After right-side injection of 10-250 pg activin protein, both early neurulae just after gastrulation movement (stage 13-14) and late neurulae just before neural tube closure (stage 17-18) showed almost 100% reversal of the heart and gut L-R axes. At higher doses of activin, more than 90% of the L-R-inverted embryos showed L-R reversal of both heart and gut. The survival ratio of the right-injected 4-day embryos was 90% on average. In the left-injected embryos, the occurrence of L-R inversion was less than 2% as observed in normal untreated siblings (1.7%). When the same amount of activin (1-50 pg) was microinjected into both sides of neurula embryos, the incidence of L-R inversion was reduced to 58%. The injection of activin along the dorsal midline in the trunk region also randomized the visceral L-R axis. Injection of activin into the right side changed normal left-handed expression of Xnr-1 to right-handed or bilateral expression. In contrast, left-handed expression of Pitx2 was switched to the right side by right activin injection. This is the first report of a method that achieves complete inversion of the visceral L-R axis by treatment of embryos at the neurula stage. Activin not only acts on the neurulae to cancel the original L R specification up to the late neurula stage, but also rebuilds a new L-R axis whose left side coincides with the injection side. It is suggested that the left and right halves of neurulae have equal potential for L-R differentiation. PMID- 10790330 TI - Induction of early transcription in one-cell mouse embryos by microinjection of the nonhistone chromosomal protein HMG-I. AB - In the mouse embryo, the onset of zygotic transcription occurs at the end of the first cell cycle, upon completion of DNA replication. We show that the nonhistone chromosomal protein HMG-I, whose translocation into the pronuclei of one-cell embryos is linked to this first round of DNA synthesis, plays a critical role in the activation of zygotic transcription. Indeed, microinjection of purified HMG-I results in a higher nuclear accumulation of the protein and triggers an earlier activation of zygotic transcription, an effect which is abolished by the preincubation of the protein with a specific antibody directed against its AT hook DNA-binding motifs. Significantly, microinjection of this antibody also prevents the normal onset of transcription in the embryo, suggesting that endogenous HMG-I is similarly involved in this process. Finally, microinjection of the exogenous protein modifies chromatin structure as measured by in situ accessibility to DNase I. We propose that general chromosomal architectural factors such as HMG-I can modulate the accessibility of chromatin to specialized regulatory factors, thereby promoting a transcriptionally competent state. PMID- 10790331 TI - Functional analysis of an eye enhancer of the Drosophila eyes absent gene: differential regulation by eye specification genes. AB - Genes involved in eye development are highly conserved between vertebrates and Drosophila. Given the complex genetic network controlling early eye development, identification of regulatory sequences controlling gene expression will provide valuable insights toward understanding central events of early eye specification. We have focused on defining regulatory elements critical for Drosophila eyes absent (eya) expression. Although eya has a complex expression pattern during development, analysis of eye-specific mutations in the gene revealed a region selectively deleted in the eye-specific alleles. Here we have performed detailed analysis of the region deleted in the eye-specific eya(2) allele. This analysis shows that this region can direct early eya gene expression in a pattern consistent with that of normal eya in eye progenitor cells. Functional studies indicate that this element will restore appropriate eya transcript expression to rescue the eye-specific allele. We have examined regulation of this element during eye specification, both in normal eye development and in ectopic eye formation. These studies demonstrate that the element was activated upon ectopic expression of the eye specification genes eyeless and dachshund, but does not respond to ectopic expression of eya or sine oculis. The differential regulation of this element by genes involved during early retinal formation reveals new aspects of the genetic hierarchy of eye development. PMID- 10790332 TI - Larval antigen molecules recognized by adult immune cells of inbred Xenopus laevis: two pathways for recognition by adult splenic T cells. AB - During anuran metamorphosis, larval cells of the tadpole are completely eliminated and replaced by adult cells in the corresponding tissues of the frog for the adaptation to terrestrial life from an aquatic life. Before the metamorphic climax, most of the cells have already transformed from larval cells into adult-type cells, but the tail cells remain as larval cells even at the climax stages of metamorphosis. In our previous works, we demonstrated that larval skin grafts are rejected by an inbred strain of adult Xenopus and that the larval cells are recognized and made apoptotic by splenocytes obtained from adults and/or metamorphosing tadpoles in vitro (Y. Izutsu and K. Yoshizato, 1993, J. Exp. Zool. 266, 163-167; Y. Izutsu et al., 1996, Differentiation 60, 277-286). In the present study, it was found that there were two types of larval epidermal cells, classified according to the presence of major histocompatibility complex (MHC); one is the apical cell expressing both MHC classes I and II, and the other is the skein cell, which expresses no MHC. By a Percoll gradient, we were able to separate these two types of cells and examined the proliferative response of adult T cells to each of them. It was revealed that the apical cells (MHC positive) were recognized directly by adult splenic T cells, whereas the skein cells (MHC-negative) were recognized by the T cells via the antigen presentation by adult splenocytes. Both of these proliferative responses were restricted to MHC class II. This is the first report showing how the larval-specific antigens present in different forms in epidermal cells are recognized as immunological targets by syngeneic adult T lymphocytes. PMID- 10790333 TI - A transient specialization of the microtubule cytoskeleton is required for differentiation of the Drosophila visual system. AB - Drosophila beta3-tubulin is an essential isoform expressed during differentiation of many cell types in embryos and pupae. We report here that during pupal development transient beta3 expression demarcates a unique subset of neurons in the developing adult visual system. beta3 is coassembled into microtubules with beta1, the sole beta-tubulin isoform in the permanent microtubule cytoskeleton of the adult eye and brain. Examination of beta3 mutant phenotypes showed that beta3 is required for axonal patterning and connectivity and for spatial positioning within the optic lobe. Comparison of the phenotypes of beta3 mutations with those that result from disruption of the Hedgehog signaling pathway shows that beta3 functions early in the establishment of the adult visual system. Our data support the hypothesis that beta3 confers specialized properties on the microtubules into which it is incorporated. Thus a transient specialization of the microtubule cytoskeleton during differentiation of a specific subset of the neurons has permanent consequences for later cell function. PMID- 10790334 TI - Spatially restricted factors cooperate with notch in the regulation of Enhancer of split genes. AB - Expression of the Drosophila Enhancer of split [E(spl)] genes, and their homologues in other species, is dependent on Notch activation. The seven E(spl) genes are clustered in a single complex and their functions overlap significantly; however, the individual genes have distinct patterns of expression. To investigate how this regulation is achieved and to find out whether there is shared or cross regulation between E(spl) genes, we have analysed the enhancer activity of sequences from the adjacent E(spl)mbeta, E(spl)mgamma and E(spl)mdelta genes and made comparisons to E(spl)m8. We find that although regulatory elements can be shared, most aspects of the expression of each individual gene are recapitulated by small (400-500 bp) evolutionarily conserved enhancers. Activated Notch or a Suppressor of Hairless-VP16 fusion are only sufficient to elicit transcription from the E(spl) enhancers in a subset of locations, indicating a requirement for other factors. In tissue culture cells, proneural proteins synergise with Suppressor of Hairless and Notch to promote expression from E(spl)mgamma and E(spl)m8, but this synergy is only observed in vivo with E(spl)m8. We conclude that additional factors besides the proneural proteins limit the response of E(spl)mgamma in vivo. In contrast to the other genes, E(spl)mbeta exhibits little response to proneural proteins and its high level of activity in the wing imaginal disc suggests that wing-specific factors cooperate with Notch to activate the E(spl)mbeta enhancer. These results demonstrate that Notch activity must be integrated with other transcriptional regulators and, since the activation of target genes is critical in determining the developmental consequences of Notch activity, provide a framework for understanding Notch function in different developmental contexts. PMID- 10790335 TI - Drosophila PTEN regulates cell growth and proliferation through PI3K-dependent and -independent pathways. AB - The control of cell and organ growth is fundamental to the development of multicellular organisms. Here, we show that dPTEN, a Drosophila homolog of the mammalian PTEN tumor suppressor gene, plays an essential role in the control of cell size, cell number, and organ size. In mosaic animals, dPTEN(-) cells proliferate faster than their heterozygous siblings, show an autonomous increase in cell size, and form organs of increased size, whereas overexpression of dPTEN results in opposite phenotypes. The loss-of-function phenotypes of dPTEN are suppressed by mutations in the PI3K target Dakt1 and the translational initiation factor eif4A, suggesting that dPTEN acts through the PI3K signaling pathway to regulate translation. Although activation of PI3K and Akt has been reported to increase rates of cellular growth but not proliferation, loss of dPTEN stimulates both of these processes, suggesting that PTEN regulates overall growth through PI3K/Akt-dependent and -independent pathways. Furthermore, we show that dPTEN does not play a major role in cell survival during Drosophila development. Our results provide a potential explanation for the high frequency of PTEN mutation in human cancer. PMID- 10790336 TI - Drosophila arc encodes a novel adherens junction-associated PDZ domain protein required for wing and eye development. AB - Loss of arc function results in a downwardly curved wing and smaller eyes with a reduced number of ommatidia. Consistent with this phenotype, molecular analysis shows that arc mRNA and protein are expressed in the wing imaginal disc and in clusters of cells in the morphogenetic furrow of the eye imaginal disc. The 36-kb arc transcription unit contains 10 exons that are spliced to form a 5. 5-kb mRNA. The encoded Arc protein is 143,000 Da and contains two PDZ (PSD-95, Discs large, ZO-1) domains; there is no close structural similarity to other PDZ proteins. In addition to its expression in imaginal discs, arc is expressed during embryogenesis in epithelia undergoing morphogenesis, including the invaginating posterior midgut, evaginating Malpighian tubule buds, elongating hindgut, invaginating salivary glands, intersegmental grooves, and developing tracheae. Arc protein colocalizes with Armadillo (beta-catenin) to the apical (luminal) surface of these developing epithelia, indicating that it is associated with adherens junctions. Genes that are required for patterning of embryonic epithelia (e.g., tailless, Kruppel, fork head, and brachyenteron) or for progression of the morphogenetic furrow (i. e., hedgehog) are required to establish or maintain the regional expression of arc. Misexpression of arc in the eye imaginal discs results in rough and larger eyes with fused ommatidia. We propose that arc affects eye development by modulating adherens junctions of the developing ommatidium. PMID- 10790337 TI - Localization of gurken RNA in Drosophila oogenesis requires elements in the 5' and 3' regions of the transcript. AB - During Drosophila oogenesis, signaling between the germline and the soma leads to the establishment of polarity in the egg and embryo. This process involves the interaction of gurken (grk), a TGFalpha-like protein, with torpedo (top), the Drosophila EGF receptor (Egfr). In early stage egg chambers, grk RNA is present predominantly along the posterior cortex of the oocyte, and in mid stage egg chambers, the grk transcript becomes tightly localized to the future dorsal anterior corner of the oocyte. This localization of grk RNA restricts the distribution of Gurken protein and is critical in defining both the anterior posterior and dorsal-ventral axes of the egg. We have determined the genomic sequence of the grk gene. By testing the requirement of various fragments of grk RNA in the localization process, we find localization signals present in both the 5' and 3' regions of the gene. Sequences in the 5' noncoding region allow for accumulation of the transcript within the oocyte in early stage egg chambers, while signals in the coding region and the 3'UTR are necessary for localization in mid to late stage egg chambers. Active translation is not required for localization of the grk RNA. The mechanism of gurken RNA localization, therefore, differs from that of other localized RNAs studied to date. PMID- 10790338 TI - : may 2, 2000 PMID- 10790339 TI - Defining diastolic dysfunction. PMID- 10790340 TI - Differential effects of the cyclin-dependent kinase inhibitors p27(Kip1), p21(Cip1), and p16(Ink4) on vascular smooth muscle cell proliferation. AB - BACKGROUND: The cyclin-dependent kinase inhibitors (CKIs) have different patterns of expression in vascular diseases. The Kip/Cip CKIs, p27(Kip1) and p21(Cip1), are upregulated during arterial repair and negatively regulate the growth of vascular smooth muscle cells (VSMCs). In contrast, the Ink CKI, p16(Ink4), is not expressed in vascular lesions. We hypothesized that a variation in the inactivation of cdk2 and cdk4 during the G(1) phase of the cell cycle by p27(Kip1), p21(Cip1), and p16(Ink4) leads to different effects on VSMC growth in vitro and in vivo. METHODS AND RESULTS: The expression of p27(Kip1) and p21(Cip1) in serum-stimulated VSMCs inactivated cdk2 and cdk4, leading to G(1) growth arrest. p16(Ink4) inhibited cdk4, but not cdk2, kinase activity, producing partial inhibition of VSMC growth in vitro. In an in vivo model of vascular injury, overexpression of p27(Kip1) reduced intimal VSMC proliferation by 52% (P<0.01) and the intima/media area ratio by 51% (P<0.005) after vascular injury and gene transfer to pig arteries, when compared with control arteries. p16(Ink4) was a weak inhibitor of intimal VSMC proliferation in injured arteries (P=NS), and it did not significantly reduce intima/media area ratios (P=NS), which is consistent with its minor effects on VSMC growth in vitro. CONCLUSIONS: p27(Kip1) and p21(Cip1) are potent inhibitors of VSMC growth compared with p16(Ink4) because of their different molecular mechanisms of cyclin-dependent kinase inhibition in the G(1) phase of the cell cycle. These findings have important implications for our understanding of the pathophysiology of vascular proliferative diseases and for the development of molecular therapies. PMID- 10790341 TI - Noninvasive, transthoracic, low-frequency ultrasound augments thrombolysis in a canine model of acute myocardial infarction. AB - BACKGROUND: Limitations of coronary thrombolysis include the time to reperfusion, patency rate, and bleeding. We evaluated the use of noninvasive transcutaneous ultrasound to augment coronary thrombolysis. METHODS AND RESULTS: In 24 dogs, a thrombotic occlusion of the left anterior descending coronary artery was induced and documented by 12-lead ECG and coronary angiography. After >/=60 minutes of occlusion, tissue-type plasminogen activator (t-PA; 1.42 mg/kg) was given intravenously over 90 minutes. A total of 12 of the 24 dogs had concomitant transcutaneous application of low-frequency ultrasound (27 kHz) over the chest. At 90 minutes, the mean TIMI grade flow in the t-PA alone group was 0.92+/-1.4 compared with 2. 42+/-1.9 in the t-PA plus ultrasound group (P=0.006). TIMI 2 to 3 flow was present in 4 of 12 cases receiving t-PA alone compared with 10 of 12 cases receiving t-PA plus ultrasound (P=0.003). At 180 minutes, mean TIMI grade flow was 0.75+/-1.4 in the t-PA alone group versus 2.58+/-0.9 in the t-PA plus ultrasound group (P=0.001). Pathological examination confirmed the angiographic patency rate and did not reveal injury secondary to ultrasound in the skin, soft tissues, heart, or lungs. CONCLUSIONS: In vivo, the noninvasive transthoracic application of low-frequency ultrasound (1) greatly augments the efficacy of t-PA mediated thrombolysis, (2) seems safe, and (3) has substantial potential as a noninvasive adjunct to improve coronary patency without increasing the risk of bleeding. PMID- 10790342 TI - Inhibition of rho-associated kinase results in suppression of neointimal formation of balloon-injured arteries. AB - BACKGROUND: Rho-associated kinase (ROCK), an effector of small GTPase Rho, regulates vascular tone via a calcium sensitization mechanism and plays a key role in the pathogenesis of hypertension. However, its role in vascular growth remains unclear. METHODS AND RESULTS: Y-27632, a specific ROCK inhibitor, and the overexpression of dominant-negative ROCK suppressed the mitogen-induced DNA synthesis of cultured vascular smooth muscle cells (VSMCs), which indicates the essential role of ROCK in the control of VSMC proliferation in vitro. Y-27632 also suppressed the chemotaxis of VSMCs. Male Wistar rats were systemically given Y-27632 (35 to 70 mg. kg(-1). day(-1)) through an intraperitoneal infusion. The neointimal formation of balloon-injured carotid arteries was significantly suppressed in Y-27632-treated rats (intima/media ratio, 0.22+/-0.02) compared with vehicle-treated rats (intima/media ratio, 0.92+/-0.21) or hydralazine treated rats with a similar blood pressure decrease (intima/media ratio, 1.03+/ 0.15). The phosphorylation of myosin phosphatase and myosin light chain was elevated in injured arteries in a Y-27632-sensitive manner, indicating the augmentation of ROCK activity in neointimal formation. The downregulation of the cyclin-dependent kinase inhibitor p27(kip1) in injured vessels was reversed by Y 27632 treatment, reflecting the antiproliferative effect of ROCK inhibition in vivo. CONCLUSIONS: We conclude that ROCK plays a key role in the process of neointimal formation after balloon injury. Thus, the inhibition of ROCK may be a potential therapeutic strategy for treating vascular proliferative disorders and hypertension. PMID- 10790343 TI - Anger proneness predicts coronary heart disease risk: prospective analysis from the atherosclerosis risk in communities (ARIC) study. AB - BACKGROUND: Increased research attention is being paid to the negative impact of anger on coronary heart disease (CHD). METHODS AND RESULTS: This study examined prospectively the association between trait anger and the risk of combined CHD (acute myocardial infarction [MI]/fatal CHD, silent MI, or cardiac revascularization procedures) and of "hard" events (acute MI/fatal CHD). Participants were 12 986 black and white men and women enrolled in the Atherosclerosis Risk In Communities study. In the entire cohort, individuals with high trait anger, compared with their low anger counterparts, were at increased risk of CHD in both event categories. The multivariate-adjusted hazard ratio (HR) (95% CI) was 1.54 (95% CI 1.10 to 2.16) for combined CHD and 1.75 (95% CI 1.17 to 2.64) for "hard" events. Heterogeneity of effect was observed by hypertensive status. Among normotensive individuals, the risk of combined CHD and of "hard" events increased monotonically with increasing levels of trait anger. The multivariate-adjusted HR of CHD for high versus low anger was 2.20 (95% CI 1.36 to 3.55) and for moderate versus low anger was 1.32 (95% CI 0.94 to 1.84). For "hard" events, the multivariate-adjusted HRs were 2.69 (95% CI 1.48 to 4.90) and 1.35 (95% CI 0.87 to 2.10), respectively. No statistically significant association between trait anger and incident CHD risk was observed among hypertensive individuals. CONCLUSIONS: Proneness to anger places normotensive middle-aged men and women at significant risk for CHD morbidity and death independent of the established biological risk factors. PMID- 10790344 TI - Type II diabetes abrogates sex differences in endothelial function in premenopausal women. AB - BACKGROUND: Obesity is a more potent cardiovascular risk factor (CVRF) in men than in women. Because traditional CVRFs cannot fully account for this sex difference, we tested the hypothesis that compared with men, women exhibit more robust endothelial function independent of obesity and that this sex difference is abrogated by diabetes. METHODS AND RESULTS: We studied leg blood flow (LBF) responses to graded intrafemoral artery infusions of the endothelium-dependent vasodilator methacholine chloride (Mch) and the endothelium-independent vasodilator sodium nitroprusside (SNP) in groups of lean, obese (OB), and type II diabetic (DM) premenopausal women and age- and body mass index-matched men. LBF response to intrafemoral administration of L-NMMA, an inhibitor of nitric oxide synthase, was also assessed in normal men and women. Maximum LBF increments in response to Mch were 347+/-57% versus 231+/-22% in lean women versus men (P<0.05) and 203+/-25% versus 111+/-17% in OB women versus men (P<0.01), respectively. In DM, maximum LBF increments in response to Mch were 104+/-24% and 138+/-33% in women and men, respectively, (P=NS). LBF decrements in response to L-NMMA were 34.9+/-4.1% and 17.1+/-4.2% in women and men, respectively (P<0.01). The response to SNP was not different between sexes and groups. CONCLUSIONS: Premenopausal nondiabetic women exhibit more robust endothelium-dependent vasodilation owing to higher rates of nitric oxide release than men. Given the protective vascular action of nitric oxide, this difference may partially explain the lower incidence of macrovascular disease in women. In premenopausal women, DM causes impairment of endothelial function beyond that observed with obesity alone and leads to endothelial dysfunction similar to that observed in DM men. These findings may help explain the similar rates of coronary artery disease and mortality in diabetic men and women. PMID- 10790345 TI - Low fasting plasma glucose level as a predictor of cardiovascular disease and all cause mortality. AB - BACKGROUND: Although medical textbooks usually classify fasting plasma glucose <70 or 80 mg/dL (<3.89 or 4.44 mmol/L) as abnormal, the prognosis for patients with low fasting plasma glucose is unclear. METHODS AND RESULTS: We conducted prospective cohort studies among 40 069 men and women to investigate the association between fasting plasma glucose levels and cardiovascular disease and all-cause mortality. We documented a U-shaped relation between fasting plasma glucose and mortality. In addition to diabetes and impaired fasting glucose levels, low fasting plasma glucose levels were also associated with high mortality. After multivariate adjustment for age, sex, study population, ethnicity, current smoking status, high blood pressure, total cholesterol, body mass index, triglycerides, history of cardiovascular disease and cancer, and a family history of cardiovascular disease, patients with fasting plasma glucose <70 mg/dL (<3.89 mmol/L) had a 3.3-fold increased risk of cardiovascular disease mortality, and patients with fasting plasma glucose 70 to 79 mg/dL (3.89 to 4.43 mmol/L) had a 2.4-fold increased risk compared with the risk in patients with fasting plasma glucose 80 to 109 mg/dL (4.44 to 6.05 mmol/L) (tests for trend P<0.0001). Participants with low fasting plasma glucose levels also had increased risk of all-cause mortality (test for trend P<0.0001). CONCLUSIONS: Participants with low fasting plasma glucose levels had a high risk of cardiovascular disease and all-cause mortality. PMID- 10790346 TI - Reducing cardiac filling pressure lowers norepinephrine spillover in patients with chronic heart failure. AB - BACKGROUND: We studied the cardiac sympathetic response to selective unloading of cardiopulmonary baroreceptors in subjects with normal left ventricular (LV) function and congestive heart failure (CHF). METHODS AND RESULTS: Eight patients with normal LV function (age 57+/-5 years, ejection fraction 58+/-2%) and 8 patients with CHF (age 60+/-2 years; ejection fraction 19+/-2%) were studied. Instrumentation consisted of an arterial line, a pulmonary artery catheter, and a coronary sinus thermodilution catheter. The radiotracer technique was used for measurement of cardiac norepinephrine spillover (CANESP) and total-body norepinephrine spillover. Lower-body negative pressure (LBNP) was applied at 2 levels: nonhypotensive and hypotensive LBNP. Nonhypotensive LBNP reduced filling pressures significantly in both groups. Arterial pressure did not change. This reduction in filling pressures caused a significant reduction in CANESP in the CHF group (from 167+/-53 to 125+/-37 pmol/min, P<0.05) but no change in the normal LV function group. Hypotensive LBNP caused a significant increase in CANESP in the normal group (73+/-13 vs 122+/-27 pmol/min, P<0.05) but no significant change in those with CHF. CONCLUSIONS: We conclude that selective reduction in filling pressures lowers cardiac norepinephrine spillover in patients with CHF. These findings suggest that a goal of CHF management should be to reduce cardiac filling pressures while avoiding systemic hypotension. PMID- 10790347 TI - Deletion allele of angiotensin-converting enzyme gene increases risk of essential hypertension in Japanese men : the Suita Study. AB - BACKGROUND: The Framingham Study recently revealed that the homozygous deletion polymorphism of the angiotensin-converting enzyme gene (ACE DD) is associated with increased risk for essential hypertension in a male-specific manner. However, this association has not been confirmed in races other than whites. METHODS AND RESULTS: Using a large number of Japanese subjects (n=5014) that were randomly selected from the general population (the Suita Study), we examined the association between ACE DD and hypertension. The frequency of DD (17.1%) in hypertensive men was significantly higher (P<0.0015) than that (11.8%) in other mildly hypertensive or normotensive men, and the estimated odds prevalence for hypertension (DD vs II) was 1.75 (95% CI 1.21 to 2.53). In contrast, no significant association was confirmed in women (OR 1.17, 95% CI 0.79 to 1.72). CONCLUSIONS: Despite the lower frequency of the DD genotype in Japanese than in whites, the ACE gene polymorphism was associated with increased risk for hypertension, suggesting that this polymorphism is a mild but certain genetic risk factor for essential hypertension in men. PMID- 10790348 TI - Improvement in exercise capacity with nitric oxide inhalation in patients with precapillary pulmonary hypertension. AB - BACKGROUND: Patients with precapillary pulmonary hypertension (PH) exhibit a poor exercise capacity due to an impaired vasodilatory response of their pulmonary arteries. By causing the pulmonary artery to dilate, inhaled nitric oxide (NO) may allow an increase in exercise capacity in patients with PH. METHODS AND RESULTS: On 2 separate days, 3 days apart, 14 patients with precapillary PH (10 primary PH, 4 residual PH after correction of an intracardiac shunt; age, 40+/-12 years; mean pulmonary artery pressure, 60+/-23 mm Hg) performed exercise, with and without inhalation of 20 ppm NO, on a cycle ergometer. The work rate was increased 15 W/min until their symptom-limited maximum, with breath-by-breath gas analysis. Patients were randomly and blindly selected to inhale NO on either their first or second test. Peak exercise load and anaerobic threshold tended to increase, but not significantly. Peak oxygen consumption (f1.gif" BORDER="0">O(2)) and Deltaf1.gif" BORDER="0">O(2)/DeltaW ratio increased significantly, by 18% and 22%, respectively (peak f1.gif" BORDER="0">O(2), 13.6+/ 3.6 to 16.0+/-4. 1 mL. kg(-1). min(-1); Deltaf1.gif" BORDER="0">O(2)/DeltaW ratio, 5. 8+/-2.4 to 7.1+/-2.3 mL. kg(-1). min(-1). W(-1); both P<0.01). Peak f1.gif" BORDER="0">O(2) increased >10% in 12 of the 14 patients. However, respiratory quotient at peak exercise decreased from 1. 22+/-0.15 to 1.09+/-0.15 (P<0.01). CONCLUSIONS: Inhaled NO substantially increases oxygen consumption at the same workload during exercise. This finding supports the possibility of ambulatory NO inhalation therapy in patients with precapillary PH. PMID- 10790349 TI - Fenfluramine and phentermine and cardiovascular findings: effect of treatment duration on prevalence of valve abnormalities. AB - BACKGROUND: The combination of fenfluramine and phentermine was a widely used obesity treatment before the withdrawal of fenfluramine for an association with heart valve regurgitation. The prevalence and clinical significance of regurgitation among patients treated with these medications has yet to be fully established. METHODS AND RESULTS: To evaluate the potential association between the duration of treatment and the prevalence of heart valve abnormalities, we examined 1163 patients who had taken fenfluramine-phentermine and 672 control patients who had not taken the drug combination within 5 years. Mild or greater aortic regurgitation was present in 8.8% of treated patients and 3.6% of control patients (P<0.001). Moderate or greater mitral regurgitation was present in 2.6% of treated patients and 1.5% of control patients (P=0.18). The adjusted odds ratio compared with controls of aortic regurgitation of mild or greater severity increased according to duration of treatment: 90 to 180 days, 1.5 (P=0.23); 181 to 360 days, 2.4 (P=0.002); 361 to 720 days, 4.6 (P<0.001); >720 days, 6.2 (P<0.001). CONCLUSIONS: This is the largest study to demonstrate a relation between the length of treatment with fenfluramine-phentermine and the prevalence of valvular abnormalities. These findings suggest that valvular abnormalities in patients who took fenfluramine-phentermine primarily involve those who had taken these medications for >6 months and predominantly results in mild aortic regurgitation. The valve regurgitation identified by this study was not accompanied by significant differences in cardiovascular symptoms nor physical findings other than a higher prevalence of heart murmurs. PMID- 10790350 TI - Electron-beam computed tomography in the assessment of coronary artery disease after heart transplantation. AB - BACKGROUND: Our aim was to compare the electron-beam CT (EBCT) features of coronary arteries in heart transplant recipients with those of biplane coronary angiography and intracoronary ultrasound (ICUS). METHODS AND RESULTS: We examined 112 heart transplant recipients (25 female; age, 17 to 69 years; median, 52 years) 1 to 153 months (median, 46 months) after surgery by EBCT to detect coronary artery calcifications. Calcifications were quantified by the Agatston scoring system. EBCT scores were compared with coronary angiography in all patients and ICUS of the left anterior descending coronary artery (LAD) in 100 patients. Coronary artery calcifications were found in 84 patients (75%). Angiographically, 16 patients displayed >50% coronary artery stenoses, all of whom had some degree of coronary artery calcification and only 1 of whom had a score of <55 (P<0.0001). With this threshold, EBCT had a sensitivity of 94%, a specificity of 79%, a positive predictive value of 43%, and a negative predictive value of 99% for detecting stenosis. ICUS confirmed the presence of calcified plaques in all patients with an LAD score >9. EBCT total calcium score was associated with the degree of intimal proliferation in that patients without ICUS features of allograft vasculopathy had a median score of 0 (25th percentile, 0; 75th percentile, 0), whereas patients with Stanford class IV vasculopathy had a median score of 41 (9 to 98, P<0.0001). CONCLUSIONS: EBCT is a promising noninvasive test for the detection of coronary heart disease in cardiac transplant recipients. PMID- 10790351 TI - Analysis of the human G protein-coupled receptor kinase 2 (GRK2) gene promoter: regulation by signal transduction systems in aortic smooth muscle cells. AB - BACKGROUND: Desensitization of G protein-coupled receptors (GPCR) is emerging as an important feature of several cardiovascular diseases. G protein-coupled receptor kinase 2 (GRK2) plays a key role in the regulation of a variety of these receptors, and its cardiac expression levels are altered in pathological situations such as chronic heart failure. However, very little is known about the signals and mechanisms that modulate GRK2 expression in cardiovascular cells. METHODS AND RESULTS: We have studied the transcriptional activity of the 1.6-kb long proximal genomic region of the human GRK2 gene. In an aortic smooth muscle cell line, agents that lead to physiological vasoconstriction and hypertrophy, such as phorbol esters, increased GRK2 promoter activity. Activation of signaling pathways by cotransfected G(alphaq) subunits or alpha(1)-adrenergic receptors also markedly enhanced the expression of the GRK2 promoter constructs. Conversely, proinflammatory cytokines, such as interleukin-1beta, tumor necrosis factor-alpha, or interferon-gamma, led to the opposite effect, decreasing the activity of the GRK2 promoter. CONCLUSIONS: Our results suggest that the expression of GRK2 in vascular cells is tightly controlled at the transcriptional level by the interplay between several extracellular messengers, which may trigger alterations of normal GRK2 levels in some physiopathological circumstances, thus promoting changes in the efficacy of the GPCR signal transduction. PMID- 10790352 TI - ATP synthesis during low-flow ischemia: influence of increased glycolytic substrate. AB - BACKGROUND: Our goals were to (1) simulate the degree of low-flow ischemia and mixed anaerobic and aerobic metabolism of an acutely infarcting region; (2) define changes in anaerobic glycolysis, oxidative phosphorylation, and the creatine kinase (CK) reaction velocity; and (3) determine whether and how increased glycolytic substrate alters the energetic profile, function, and recovery of the ischemic myocardium in the isolated blood-perfused rat heart. METHODS AND RESULTS: Hearts had 60 minutes of low-flow ischemia (10% of baseline coronary flow) and 30 minutes of reperfusion with either control or high glucose and insulin (G+I) as substrate. In controls, during ischemia, rate-pressure product and oxygen consumption decreased by 84%. CK velocity decreased by 64%; ATP and phosphocreatine (PCr) concentrations decreased by 51% and 63%, respectively; inorganic phosphate (P(i)) concentration increased by 300%; and free [ADP] did not increase. During ischemia, relative to controls, the G+I group had similar CK velocity, oxygen consumption, and tissue acidosis but increased glycolysis, higher [ATP] and [PCr], and lower [P(i)] and therefore had a greater free energy yield from ATP hydrolysis. Ischemic systolic and diastolic function and postischemic recovery were better. CONCLUSIONS: During low-flow ischemia simulating an acute myocardial infarction region, oxidative phosphorylation accounted for 90% of ATP synthesis. The CK velocity fell by 66%, and CK did not completely use available PCr to slow ATP depletion. G+I, by increasing glycolysis, slowed ATP depletion, maintained lower [P(i)], and maintained a higher free energy from ATP hydrolysis. This improved energetic profile resulted in better systolic and diastolic function during ischemia and reperfusion. These results support the clinical use of G+I in acute MI. PMID- 10790353 TI - Endothelin-1 vasoconstriction during swine cardiopulmonary resuscitation improves coronary perfusion pressures but worsens postresuscitation outcome. AB - BACKGROUND: Vasoconstriction during cardiopulmonary resuscitation (CPR) improves coronary perfusion pressure (CPP) and thereby outcome. The combination of endothelin-1 (ET-1) plus epinephrine improved CPP during CPR compared with epinephrine alone in a canine cardiac arrest model. The effect of the combination on outcome variables, such as successful resuscitation and survival, has not been investigated. METHODS AND RESULTS: Twenty-seven swine were randomly provided with 1 mg epinephrine (Epi group) or 1 mg epinephrine plus 0.1 mg ET-1 (ET-1 group) during a prolonged ventricular fibrillatory cardiac arrest. ET-1 resulted in substantially superior aortic relaxation pressure and CPP during CPR. These hemodynamic improvements tended to increase initial rates of restoration of spontaneous circulation (8 of 10 versus 8 of 17, P=0.12). However, continued intense vasoconstriction from ET-1 led to higher aortic diastolic pressure and very narrow pulse pressure after resuscitation. The mean pulse pressure 1 hour after resuscitation was 7+/-8 mm Hg with ET-1 versus 24+/-1 mm Hg with Epi, P<0.01. Most importantly, the postresuscitation mortality was dramatically higher in the ET-1 group (6 of 8 versus 0 of 8 in the Epi group, P<0.01). CONCLUSIONS: These data establish that administration of ET-1 during CPR can result in worse postresuscitation outcome. The intense vasoconstriction from ET-1 improved CPP during CPR but had detrimental effects in the postresuscitation period. PMID- 10790354 TI - beta-adrenergic blockade in developing heart failure: effects on myocardial inflammatory cytokines, nitric oxide, and remodeling. AB - BACKGROUND: Whether beta-adrenergic blockade modulates myocardial expression of inflammatory cytokines and nitric oxide (NO) in heart failure is unclear. METHODS AND RESULTS: We administered oral metoprolol or no therapy to rats for 12 weeks after large myocardial infarction and subsequently examined left ventricular (LV) remodeling; myocardial tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 expression; and NO. In untreated rats, echocardiography revealed significant (P<0.001) LV dilatation and systolic dysfunction compared with sham. Papillary muscle studies revealed isoproterenol hyporesponsiveness to be unaltered by NO synthase (NOS) inhibition. Circulating NO metabolites were undetectable. In noninfarcted myocardium, although inducible NOS (iNOS) mRNA was absent, TNF-alpha, IL-1beta, and IL-6 mRNA and protein were markedly elevated compared with sham (P<0.001), with 2-fold higher expression (P<0.025) of IL-6 compared with TNF-alpha or IL-1beta. Metoprolol administration starting 48 hours after infarction (1) attenuated (P<0.02) LV dilatation and systolic dysfunction, (2) preserved isoproterenol responsiveness (P<0.025) via NO-independent mechanisms, and (3) reduced myocardial gene expression and protein production of TNF-alpha and IL-1beta (P<0. 025) but not IL-6, which remained high. CONCLUSIONS: During heart failure development, adrenergic activation contributes to increased myocardial expression of TNF-alpha and IL-1beta but not IL-6, and one mechanism underlying the beneficial effects of beta-adrenergic blockade may involve attenuation of TNF-alpha and IL-1beta expression independent of iNOS and NO. PMID- 10790355 TI - Surface potentials from the region of the atrioventricular node and their relation to dual pathway electrophysiology. AB - BACKGROUND: Clinical applications of the principles of dual atrioventricular nodal (AVN) electrophysiology in the treatment of AVN reentrant tachycardias rely on empirical findings, such as discontinued conduction curves or the presence of specific catheter-recorded signals. However, neither the shape of the conduction curve nor the surface electrograms have been validated as functionally related to the presence of slow or fast wavefronts. METHODS AND RESULTS: We performed in vitro studies using 10 rabbit atrial-AVN preparations. A bipolar roving electrode was used to explore the endocardial surface of the triangle of Koch during programmed electrical stimulation. Microelectrodes were impaled in AVN cells to correlate surface and intracellular responses. In 7 preparations, a specific area near the compact cell region produced surface electrograms that were dissociated in 2 distinct components, with progressive shortening of prematurity. Similar dissociation was demonstrated during Wenckebach periodicity and increased vagal tone. Cellular recordings supported the presence of early ("fast") and late ("slow") wavefronts, with different refractory properties. Although the fast-slow transition was a basis for discontinued propagation, the AVN conduction curves were smooth in the majority of cases. CONCLUSIONS: Exploration of the triangle of Koch during programmed pacing reveals the presence of dual-wavefront surface potentials. Clinical confirmation of these AVN potentials could provide a new, sensitive tool in defining dual AVN electrophysiology. PMID- 10790356 TI - Defining diastolic heart failure: a call for standardized diagnostic criteria. PMID- 10790357 TI - Images in Cardiovascular Medicine. Neurogenic stunned myocardium. PMID- 10790358 TI - Primary cardiac lymphoma diagnosed with transesophageal echocardiography-guided endomyocardial biopsy. PMID- 10790359 TI - EMBO WORKSHOP REPORT: Molecular and cellular gerontology Serpiano, Switzerland, September 18-22, 1999. PMID- 10790360 TI - EMBO Workshop Report: Molecular genetics of muscle development and neuromuscular diseases Kloster Irsee, Germany, September 26-October 1, 1999. PMID- 10790361 TI - Dependence and independence of [PSI(+)] and [PIN(+)]: a two-prion system in yeast? AB - The [PSI(+)] prion can be induced by overproduction of the complete Sup35 protein, but only in strains carrying the non-Mendelian [PIN(+)] determinant. Here we demonstrate that just as [psi (-)] strains can exist as [PIN(+)] and [pin(-)] variants, [PSI(+)] can also exist in the presence or absence of [PIN(+)]. [PSI(+)] and [PIN(+)] tend to be cured together, but can be lost separately. [PSI(+)]-related phenotypes are not affected by [PIN(+)]. Thus, [PIN(+)] is required for the de novo formation of [PSI(+)], not for [PSI(+)] propagation. Although [PSI(+)] induction is shown to require [PIN(+)] even when the only overexpressed region of Sup35p is the prion domain, two altered prion domain fragments circumventing the [PIN(+)] requirement are characterized. Finally, in strains cured of [PIN(+)], prolonged incubation facilitates the reappearance of [PIN(+)]. Newly appearing [PIN(+)] elements are often unstable but become stable in some mitotic progeny. Such reversibility of curing, together with our previous demonstration that the inheritance of [PIN(+)] is non Mendelian, supports the hypothesis that [PIN(+)] is a prion. Models for [PIN(+)] action, which explain these findings, are discussed. PMID- 10790362 TI - Lipophosphoglycan is not required for infection of macrophages or mice by Leishmania mexicana. AB - Cell surface lipophosphoglycan (LPG) is commonly regarded as a multifunctional Leishmania virulence factor required for survival and development of these parasites in mammals. In this study, the LPG biosynthesis gene lpg1 was deleted in Leishmania mexicana by targeted gene replacement. The resulting mutants are deficient in LPG synthesis but still display on their surface and secrete phosphoglycan-modified molecules, most likely in the form of proteophosphoglycans, whose expression appears to be up-regulated. LPG-deficient L.mexicana promastigotes show no significant differences to LPG-expressing parasites with respect to attachment to, uptake into and multiplication inside macrophages. Moreover, in Balb/c and C57/BL6 mice, LPG-deficient L.mexicana clones are at least as virulent as the parental wild-type strain and lead to lethal disseminated disease. The results demonstrate that at least L. mexicana does not require LPG for experimental infections of macrophages or mice. Leishmania mexicana LPG is therefore not a virulence factor in the mammalian host. PMID- 10790363 TI - MEF2 responds to multiple calcium-regulated signals in the control of skeletal muscle fiber type. AB - Different patterns of motor nerve activity drive distinctive programs of gene transcription in skeletal muscles, thereby establishing a high degree of metabolic and physiological specialization among myofiber subtypes. Recently, we proposed that the influence of motor nerve activity on skeletal muscle fiber type is transduced to the relevant genes by calcineurin, which controls the functional activity of NFAT (nuclear family of activated T cell) proteins. Here we demonstrate that calcineurin-dependent gene regulation in skeletal myocytes is mediated also by MEF2 transcription factors, and is integrated with additional calcium-regulated signaling inputs, specifically calmodulin-dependent protein kinase activity. In skeletal muscles of transgenic mice, both NFAT and MEF2 binding sites are necessary for properly regulated function of a slow fiber specific enhancer, and either forced expression of activated calcineurin or motor nerve stimulation up-regulates a MEF2-dependent reporter gene. These results provide new insights into the molecular mechanisms by which specialized characteristics of skeletal myofiber subtypes are established and maintained. PMID- 10790364 TI - A putative endosomal t-SNARE links exo- and endocytosis in the phytopathogenic fungus Ustilago maydis. AB - We identified a temperature-sensitive mutant of the plant pathogenic fungus Ustilago maydis that is defective in the polar distribution of cell wall components and shows abnormal morphology. The affected gene, yup1, was cloned by complementation. It encodes a putative target soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (t-SNARE), suggesting a function in membrane fusion. A Yup1-GFP fusion protein localized to vesicles that showed rapid saltatory motion along microtubules. These vesicles are part of the endocytic pathway and accumulate at sites of active growth, thereby supporting the expansion of the hyphal tip. In yup1(ts) cells, endocytosis is impaired and accumulation of Yup1-carrying endosomes at cell poles is abolished, resulting in apolar distribution of wall components and morphological alterations. This suggests that a membrane recycling process via early endosomes supports polar growth of U. maydis. PMID- 10790365 TI - MafB is an inducer of monocytic differentiation. AB - The bZip transcription factor MafB is expressed specifically in the myeloid lineage of the hematopoietic system and is up-regulated successively during myeloid differentiation from multipotent progenitors to macrophages. Here we report that this induction reflects an essential role of MafB in early myeloid and monocytic differentiation. We observed that the expression of MafB in transformed chicken hematopoietic precursors dramatically increases the proportion of myeloid colony formation at the expense of multipotent progenitor type colonies. In addition, the overexpression of MafB in transformed myeloblasts stimulates the rapid formation of macrophages, as judged by morphology, surface marker expression and functional criteria. MafB-induced macrophages exhibit typical levels of phagocytic activity and nitric oxide release after activation by lipopolysaccharide. By contrast, overexpression of the myeloid transcription factor PU.1 in these cells does not induce macrophage differentiation. Furthermore, a dominant-negative allele of MafB inhibits both myeloid colony formation and the differentiation of myeloblasts into macrophages. Taken together, our results indicate that MafB induction is a specific and essential determinant of the monocytic program in hematopoietic cells. PMID- 10790367 TI - Adhesion to the extracellular matrix regulates the coupling of the small GTPase Rac to its effector PAK. AB - The small GTPase Rac regulates cytoskeletal organization, cell cycle progression, gene expression and oncogenic transformation, processes that depend upon both soluble growth factors and adhesion to the extracellular matrix (ECM). We now show that growth factors and adhesion to the ECM both contribute independently and approximately equally to Rac activation. However, activated Rac in non adherent cells failed to stimulate the Rac effector PAK. V12 Rac or Rac activated by serum translocated to the membrane fraction of adherent cells but remained mainly cytoplasmic in suspended cells. An activated Rac mutant lacking a membrane targeting sequence did not activate PAK in adherent cells, while mutations that forced membrane targeting restored PAK activation in suspended cells. In vitro, V12 Rac showed greater binding to membranes from adherent relative to suspended cells, indicating that cell adhesion regulated membrane binding sites for Rac. These results show that ECM regulates the ability of Rac to couple with PAK via an effect on membrane binding sites that facilitate their interaction. PMID- 10790366 TI - The Id4 HLH protein and the timing of oligodendrocyte differentiation. AB - An intracellular timer is thought to help control the timing of oligodendrocyte differentiation. We show here that the expression of the helix-loop-helix gene Id4 in oligodendrocyte precursor cells decreases in vivo and in vitro with a time course expected if Id4 is part of the timer. We also show that Id4 expression decreases prematurely when the precursor cells are induced to differentiate by mitogen withdrawal. Both Id4 mRNA and protein decrease together under all of these conditions, suggesting that the control of Id4 expression is transcriptional. Finally, we show that enforced expression of Id4 stimulates cell proliferation and blocks differentiation induced by either mitogen withdrawal or treatment with thyroid hormone. These findings suggest that a progressive fall in Id4 transcription is part of the intracellular timer that helps determine when oligodendrocyte precursor cells withdraw from the cell cycle and differentiate. PMID- 10790368 TI - NKX2.3 is required for MAdCAM-1 expression and homing of lymphocytes in spleen and mucosa-associated lymphoid tissue. AB - Targeted disruption of the transcription factor NKX2.3 gene in mice results in anatomical defects of intestine and secondary lymphoid organs. Here, we report that spleen and Peyer's patches of NKX2. 3-deficient mice are considerably reduced in size and lack the ordered tissue architecture. T and B cells are misplaced within the spleen and mesenteric lymph nodes and fail to segregate into the appropriate T and B cell areas. Furthermore, splenic marginal zones, characterized by specific B cells and various types of macrophage-derived cells around the marginal sinus, are absent in mutants. Homozygous NKX2.3 mutants lack the mucosal addressin cell adhesion molecule-1 (MAdCAM-1) that is normally expressed in mucosa-associated lymphoid tissue (MALT) and spleen. We provide evidence that NKX2.3 can activate MAdCAM-1 transcription directly, suggesting that MAdCAM-1 is at least partly responsible for the migration and homing defects of lymphocytes and macrophages in mutants. Therefore, expression of MAdCAM-1 seems to be required for building functional structures in spleen and MALT, a prerequisite for unimpaired migration and segregation of B and T cells to and within these organs. PMID- 10790369 TI - The tight junction protein ZO-1 and an interacting transcription factor regulate ErbB-2 expression. AB - Epithelial tight junctions regulate paracellular diffusion and restrict the intermixing of apical and basolateral plasma membrane components. We now identify a Y-box transcription factor, ZONAB (ZO-1-associated nucleic acid-binding protein), that binds to the SH3 domain of ZO-1, a submembrane protein of tight junctions. ZONAB localizes to the nucleus and at tight junctions, and binds to sequences of specific promoters containing an inverted CCAAT box. In reporter assays, ZONAB and ZO-1 functionally interact in the regulation of the ErbB-2 promoter in a cell density-dependent manner. In stably transfected overexpressing cells, ZO-1 and ZONAB control expression of endogenous ErbB-2 and function in the regulation of paracellular permeability. These data indicate that tight junctions directly participate in the control of gene expression and suggest that they function in the regulation of epithelial cell differentiation. PMID- 10790370 TI - TCRalpha enhancer activation occurs via a conformational change of a pre assembled nucleo-protein complex. AB - The TCR alpha enhancer (Ealpha) has served as a paradigm for studying how enhancers organize trans-activators into nucleo-protein complexes thought to recruit and synergistically stimulate the transcriptional machinery. Little is known, however, of either the extent or dynamics of Ealpha occupancy by nuclear factors during T cell development. Using dimethyl sulfate (DMS) in vivo footprinting, we demonstrate extensive Ealpha occupancy, encompassing both previously identified and novel sites, not only in T cells representing a developmental stage where Ealpha is known to be active (CD4(+)CD8(+)-DP cells), but surprisingly, also in cells at an earlier developmental stage where Ealpha is not active (CD4(-)CD8(-)-DN cells). Partial occupancy was also established in B lymphoid but not non-lymphoid cells. In vivo DNase I footprinting, however, implied developmentally induced changes in nucleo-protein complex topography. Stage-specific differences in factor composition at Ealpha sequences were also suggested by EMSA analysis. These results, which indicate that alterations in the structure of a pre-assembled nucleo-protein complex correlate with the onset of Ealpha activity, may exemplify one mechanism by which enhancers can rapidly respond to incoming stimuli. PMID- 10790371 TI - GATA-dependent recruitment of MEF2 proteins to target promoters. AB - The myocyte enhancer factor-2 (MEF2) proteins are MADS-box transcription factors that are essential for differentiation of all muscle lineages but their mechanisms of action remain largely undefined. In mammals, the earliest site of MEF2 expression is the heart where the MEF2C isoform is detectable as early as embryonic day 7.5. Inactivation of the MEF2C gene causes cardiac developmental arrest and severe downregulation of a number of cardiac markers including atrial natriuretic factor (ANF). However, most of these promoters contain no or low affinity MEF2 binding sites and they are not significantly activated by any MEF2 proteins in heterologous cells suggesting a dependence on a cardiac-enriched cofactor for MEF2 action. We provide evidence that MEF2 proteins are recruited to target promoters by the cell-specific GATA transcription factors, and that MEF2 potentiates the transcriptional activity of this family of tissue-restricted zinc finger proteins. Functional MEF2/GATA-4 synergy involves physical interaction between the MEF2 DNA-binding domain and the carboxy zinc finger of GATA-4 and requires the activation domains of both proteins. However, neither MEF2 binding sites nor MEF2 DNA binding capacity are required for transcriptional synergy. The results unravel a novel pathway for transcriptional regulation by MEF2 and provide a molecular paradigm for elucidating the mechanisms of action of MEF2 in muscle and non-muscle cells. PMID- 10790372 TI - Cell cycle-dependent variations in c-Jun and JunB phosphorylation: a role in the control of cyclin D1 expression. AB - The transcription factor AP-1, composed of Jun and Fos proteins, is a major target of mitogen-activated signal transduction pathways. However, little is known about AP-1 function in normal cycling cells. Here we report that the quantity and the phosphorylation state of the c-Jun and JunB proteins vary at the M-G(1) transition. Phosphorylation of JunB by the p34(cdc2)-cyclin B kinase is associated with lower JunB protein levels in mitotic and early G(1) cells. In contrast, c-Jun levels remain constant while the protein undergoes N-terminal phosphorylation, increasing its transactivation potential. Since JunB represses and c-Jun activates the cyclin D1 promoter, these modifications of AP-1 activity during the M-G(1) transition could provide an impetus for G(1) progression by a temporal increase in cyclin D1 transcription. These findings constitute a novel example of a reciprocal connection between transcription factors and the cell cycle machinery. PMID- 10790373 TI - Targeted disruption of Skp2 results in accumulation of cyclin E and p27(Kip1), polyploidy and centrosome overduplication. AB - The ubiquitin-proteasome pathway plays an important role in control of the abundance of cell cycle regulators. Mice lacking Skp2, an F-box protein and substrate recognition component of an Skp1-Cullin-F-box protein (SCF) ubiquitin ligase, were generated. Although Skp2(-/-) animals are viable, cells in the mutant mice contain markedly enlarged nuclei with polyploidy and multiple centrosomes, and show a reduced growth rate and increased apoptosis. Skp2(-/-) cells also exhibit increased accumulation of both cyclin E and p27(Kip1). The elimination of cyclin E during S and G(2) phases is impaired in Skp2(-/-) cells, resulting in loss of cyclin E periodicity. Biochemical studies showed that Skp2 interacts specifically with cyclin E and thereby promotes its ubiquitylation and degradation both in vivo and in vitro. These results suggest that specific degradation of cyclin E and p27(Kip1) is mediated by the SCF(Skp2) ubiquitin ligase complex, and that Skp2 may control chromosome replication and centrosome duplication by determining the abundance of cell cycle regulators. PMID- 10790375 TI - Measuring motion on DNA by the type I restriction endonuclease EcoR124I using triplex displacement. AB - The type I restriction enzyme EcoR124I cleaves DNA following extensive linear translocation dependent upon ATP hydrolysis. Using protein-directed displacement of a DNA triplex, we have determined the kinetics of one-dimensional motion without the necessity of measuring DNA or ATP hydrolysis. The triplex was pre formed specifically on linear DNA, 4370 bp from an EcoR124I site, and then incubated with endonuclease. Upon ATP addition, a distinct lag phase was observed before the triplex-forming oligonucleotide was displaced with exponential kinetics. As the distance between type I and triplex sites was shortened, the lag time decreased whilst the displacement reaction remained exponential. This is indicative of processive DNA translocation followed by collision with the triplex and oligonucleotide displacement. A linear relationship between lag duration and inter-site distance gives a translocation velocity of 400+/-32 bp/s at 20 degrees C. Furthermore, the data can only be explained by bi-directional translocation. An endonuclease with only one of the two HsdR subunits responsible for motion could still catalyse translocation. The reaction is less processive, but can 'reset' in either direction whenever the DNA is released. PMID- 10790374 TI - DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p. AB - Cdc45p assembles at replication origins before initia tion and is required for origin firing in Saccharomyces cerevisiae. A heat-inducible cdc45 degron mutant was constructed that promotes rapid degradation of Cdc45p at the restrictive temperature. Consistent with a role in initiation, loss of Cdc45p in G(1) prevents all detectable DNA replication without preventing subsequent entry into mitosis. Loss of Cdc45p activity during S-phase blocks S-phase completion but not activation of replication checkpoints. Using density substitution, we show that after allowing replication fork establishment, Cdc45p inactivation prevents the subsequent progression of individual replication forks. This provides the first direct functional evidence that Cdc45p plays an essential role during elongation. Thus, like the large T antigen in SV40 replication, Cdc45p plays a central role in both initiation and elongation phases of chromosomal DNA replication. PMID- 10790376 TI - Hybrid mouse-prokaryotic DNA (cytosine-5) methyltransferases retain the specificity of the parental C-terminal domain. AB - The mouse (cytosine-5) DNA methyltransferase (Dnmt1) consists of a regulatory N terminal and a catalytic C-terminal domain, which are fused by a stretch of Gly Lys dipeptide repeats. The C-terminal region contains all of the conserved motifs found in other cytosine-5 DNA methyltransferases including the relative position of the catalytic Pro-Cys dipeptide. In prokaryotes, the methyltransferases are simpler and lack the regulatory N-terminal domain. We constructed three hybrid methyltransferases, containing the intact N-terminus of the murine Dnmt1 and most of the coding sequences from M.HhaI (GCGC), M.HpaII (CCGG) or M.SssI (CG). These hybrids are biologically active when expressed in a baculovirus system and show the specificity of the parental C-terminal domain. Expression of these recombinant constructs leads to de novo methylation of both host and viral genomes in a sequence-specific manner. Steady-state kinetic analyses were performed on the murine Dnmt1-HhaI hybrid using poly(dG-dC).poly (dG-dC), unmethylated and hemimethylated oligonucleotides as substrates. The enzyme has a slow catalytic turnover number of 4.38 h(-1) for poly(dG-dC). poly(dG-dC), and exhibits 3-fold higher catalytic efficiency for hemimethylated substrates. PMID- 10790377 TI - Rcl1p, the yeast protein similar to the RNA 3'-phosphate cyclase, associates with U3 snoRNP and is required for 18S rRNA biogenesis. AB - RNA 3'-terminal phosphate cyclases are evolutionarily conserved enzymes catalysing conversion of the 3'-terminal phosphate in RNA to the 2',3'-cyclic phosphodiester. Their biological role remains unknown. The yeast Saccharomyces cerevisiae contains a gene encoding a protein with strong sequence similarity to the characterized cyclases from humans and Escherichia coli. The gene, named RCL1 (for RNA terminal phosphate cyclase like), is essential for growth, and its product, Rcl1p, is localized in the nucleolus. Depletion or inactivation of Rcl1p impairs pre-rRNA processing at sites A(0), A(1) and A(2), and leads to a strong decrease in 18S rRNA and 40S ribosomal subunit levels. Immunoprecipitations indicate that Rcl1p is specifically associated with the U3 snoRNP, although, based on gradient analyses, it is not its structural component. Most of Rcl1p sediments in association with the 70-80S pre-ribosomal particle and a 10S complex of unknown identity. Proteins similar to Rcl1p are encoded in genomes of all eukaryotes investigated and the mouse orthologue complements yeast strains depleted of Rcl1p. Possible functions of Rcl1p in pre-rRNA processing and its relationship to the RNA 3'-phosphate cyclase are discussed. PMID- 10790379 TI - Kay Wilson and the NIH genetics study section. PMID- 10790378 TI - Late events of translation initiation in bacteria: a kinetic analysis. AB - Binding of the 50S ribosomal subunit to the 30S initiation complex and the subsequent transition from the initiation to the elongation phase up to the synthesis of the first peptide bond represent crucial steps in the translation pathway. The reactions that characterize these transitions were analyzed by quench-flow and fluorescence stopped-flow kinetic techniques. IF2-dependent GTP hydrolysis was fast (30/s) followed by slow P(i) release from the complex (1.5/s). The latter step was rate limiting for subsequent A-site binding of EF-Tu small middle dotGTP small middle dotPhe-tRNA(Phe) ternary complex. Most of the elemental rate constants of A-site binding were similar to those measured on poly(U), with the notable exception of the formation of the first peptide bond which occurred at a rate of 0.2/s. Omission of GTP or its replacement with GDP had no effect, indicating that neither the adjustment of fMet-tRNA(fMet) in the P site nor the release of IF2 from the ribosome required GTP hydrolysis. PMID- 10790380 TI - Negligible genetic diversity of mycobacterium tuberculosis host immune system protein targets: evidence of limited selective pressure. AB - A common theme in medical microbiology is that the amount of amino acid sequence variation in proteins that are targets of the host immune system greatly exceeds that found in metabolic enzymes or other housekeeping proteins. Twenty-four Mycobacterium tuberculosis genes coding for targets of the host immune system were sequenced in 16 strains representing the breadth of genomic diversity in the species. Of the 24 genes, 19 were invariant and only six polymorphic nucleotide sites were identified in the 5 genes that did have variation. The results document the highly unusual circumstance that prominent M. tuberculosis antigenic proteins have negligible structural variation worldwide. The data are best explained by a combination of three factors: (i) evolutionarily recent global dissemination in humans, (ii) lengthy intracellular quiescence, and (iii) active replication in relatively few fully immunocompetent hosts. The very low level of amino acid diversity in antigenic proteins may be cause for optimism in the difficult fight to control global tuberculosis. PMID- 10790381 TI - A STE12 homolog is required for mating but dispensable for filamentation in candida lusitaniae. AB - Candida lusitaniae is a dimorphic yeast that is emerging as an opportunistic fungal pathogen. In contrast to Candida albicans, which is diploid and asexual, C. lusitaniae has been reported to have a sexual cycle. We have employed genetic approaches to demonstrate that C. lusitaniae is haploid and has a sexual cycle involving mating between MATa and MATalpha cells under nutrient deprivation conditions. By degenerate PCR, we identified a C. lusitaniae homolog (Cls12) of the Ste12 transcription factor that regulates mating, filamentation, and virulence in Saccharomyces cerevisiae, C. albicans, and Cryptococcus neoformans. Comparison of the CLS12 DNA and protein sequences to other STE12 homologs and transformation experiments with selectable markers from S. cerevisiae (URA3, KanMX, HphMX) and C. albicans (CaURA3) provide evidence that the CUG codon encodes serine instead of leucine in C. lusitaniae, as is also the case in C. albicans. The C. lusitaniae CLS12 gene was disrupted by biolistic transformation and homologous recombination. C. lusitaniae cls12 mutant strains were sterile but had no defect in filamentous growth. Our findings reveal both conserved and divergent roles for the C. lusitaniae STE12 homolog in regulating differentiation of this emerging fungal pathogen. PMID- 10790382 TI - Inhibition of mRNA turnover in yeast by an xrn1 mutation enhances the requirement for eIF4E binding to eIF4G and for proper capping of transcripts by Ceg1p. AB - Null mutants of XRN1, encoding the major cytoplasmic exoribonuclease in yeast, are viable but accumulate decapped, deadenylated transcripts. A screen for mutations synthetic lethal with xrn1Delta identified a mutation in CDC33, encoding eIF4E. This mutation (glutamate to glycine at position 72) affected a highly conserved residue involved in interaction with eIF4G. Synthetic lethality between xrn1 and cdc33 was not relieved by high-copy expression of eIF4G or by disruption of the yeast eIF4E binding protein Caf20p. High-copy expression of a mutant eIF4G defective for eIF4E binding resulted in a dominant negative phenotype in an xrn1 mutant, indicating the importance of this interaction in an xrn1 mutant. Another allele of CDC33, cdc33-1, along with mutations in CEG1, encoding the nuclear guanylyltransferase, were also synthetic lethal with xrn1Delta, whereas mutations in PRT1, encoding a subunit of eIF3, were not. Mutations in CDC33, CEG1, PRT1, PAB1, and TIF4631, encoding eIF4G1, have been shown to lead to destabilization of mRNAs. Although such destabilization in cdc33, ceg1, and pab1 mutants can be partially suppressed by an xrn1 mutation, we observed synthetic lethality between xrn1 and either cdc33 or ceg1 and no suppression of the inviability of a pab1 null mutation by xrn1Delta. Thus, the inhibition of mRNA turnover by blocking Xrn1p function does not suppress the lethality of defects upstream in the turnover pathway but it does enhance the requirement for (7)mG caps and for proper formation of the eIF4E/eIF4G cap recognition complex. PMID- 10790383 TI - Point mutations identify a conserved region of the saccharomyces cerevisiae AFR1 gene that is essential for both the pheromone signaling and morphogenesis functions. AB - Mating pheromone receptors activate a G protein signal pathway that leads to the conjugation of the yeast Saccharomyces cerevisiae. This pathway also induces the production of Afr1p, a protein that negatively regulates pheromone receptor signaling and is required to form pointed projections of new growth that become the site of cell fusion during mating. Afr1p lacks strong similarity to any well characterized proteins to help predict how it acts. Therefore, we investigated the relationship between the different functions of Afr1p by isolating and characterizing seven mutants that were defective in regulating pheromone signaling. The AFR1 mutants were also defective when expressed as fusions to STE2, the alpha-factor receptor, indicating that the mutant Afr1 proteins are defective in function and not in co-localizing with receptors. The mutant genes contained four distinct point mutations that all occurred between codons 254 and 263, identifying a region that is critical for AFR1 function. Consistent with this, we found that the corresponding region is very highly conserved in the Afr1p homologs from the yeasts S. uvarum and S. douglasii. In contrast, there were no detectable effects on pheromone signaling caused by deletion or overexpression of YER158c, an open reading frame with overall sequence similarity to Afr1p that lacks this essential region. Interestingly, all of the AFR1 mutants showed a defect in their ability to form mating projections that was proportional to their defect in regulating pheromone signaling. This suggests that both functions may be due to the same action of Afr1p. Thus, these studies identify a specific region of Afr1p that is critical for its function in both signaling and morphogenesis. PMID- 10790384 TI - TUP1, CPH1 and EFG1 make independent contributions to filamentation in candida albicans. AB - The common fungal pathogen, Candida albicans, can grow either as single cells or as filaments (hyphae), depending on environmental conditions. Several transcriptional regulators have been identified as having key roles in controlling filamentous growth, including the products of the TUP1, CPH1, and EFG1 genes. We show, through a set of single, double, and triple mutants, that these genes act in an additive fashion to control filamentous growth, suggesting that each gene represents a separate pathway of control. We also show that environmentally induced filamentous growth can occur even in the absence of all three of these genes, providing evidence for a fourth regulatory pathway. Expression of a collection of structural genes associated with filamentous growth, including HYR1, ECE1, HWP1, ALS1, and CHS2, was monitored in strains lacking each combination of TUP1, EFG1, and CPH1. Different patterns of expression were observed among these target genes, supporting the hypothesis that these three regulatory proteins engage in a network of individual connections to downstream genes and arguing against a model whereby the target genes are regulated through a central filamentous growth pathway. The results suggest the existence of several distinct types of filamentous forms of C. albicans, each dependent on a particular set of environmental conditions and each expressing a unique set of surface proteins. PMID- 10790385 TI - Genetic interactions between GLC7, PPZ1 and PPZ2 in saccharomyces cerevisiae. AB - GLC7 encodes an essential serine/threonine protein type I phosphatase in Saccharomyces cerevisiae. Three other phosphatases (Ppz1p, Ppz2p, and Sal6p) share >59% identity in their catalytic region with Glc7p. ppz1 ppz2 null mutants have no apparent growth defect on rich media. However, null alleles of PPZ1 and PPZ2, in combination with mutant alleles of GLC7, confer a range of growth defects varying from slow growth to lethality. These results indicate that Glc7p, Ppz1p, and Ppz2p may have overlapping functions. To determine if this overlap extends to interaction with targeting subunits, Glc7p-binding proteins were tested for interaction in the two-hybrid system with the functional catalytic domain of Ppz1p. Ppz1p interacts strongly with a number of Glc7p regulatory subunits, including Glc8p, a protein that shares homology with mammalian PP1 inhibitor I2. Genetic data suggest that Glc8p positively affects both Glc7p and Ppz1p functions. Together our data suggest that Ppz1p and Ppz2p may have overlapping functions with Glc7p and that all three phosphatases may act through common regulatory proteins. PMID- 10790386 TI - A transmembrane guanylyl cyclase (DAF-11) and Hsp90 (DAF-21) regulate a common set of chemosensory behaviors in caenorhabditis elegans. AB - Caenorhabditis elegans daf-11 and daf-21 mutants share defects in specific chemosensory responses mediated by several classes of sensory neurons, indicating that these two genes have closely related functions in an assortment of chemosensory pathways. We report that daf-11 encodes one of a large family of C. elegans transmembrane guanylyl cyclases (TM-GCs). The cyclic GMP analogue 8-bromo cGMP rescues a sensory defect in both daf-11 and daf-21 mutants, supporting a role for DAF-11 guanylyl cyclase activity in this process and further suggesting that daf-21 acts at a similar step. daf-11::gfp fusions are expressed in five identified pairs of chemosensory neurons in a pattern consistent with most daf-11 mutant phenotypes. We also show that daf-21 encodes the heat-shock protein 90 (Hsp90), a chaperone with numerous specific protein targets. We show that the viable chemosensory-deficient daf-21 mutation is an unusual allele resulting from a single amino acid substitution and that the daf-21 null phenotype is early larval lethality. These results demonstrate that cGMP is a prominent second messenger in C. elegans chemosensory transduction and suggest a previously unknown role for Hsp90 in regulating cGMP levels. PMID- 10790388 TI - Molecular evolution of odorant-binding protein genes OS-E and OS-F in Drosophila. AB - The Drosophila olfactory genes OS-E and OS-F are members of a family of genes that encode insect odorant-binding proteins (OBPs). OBPs are believed to transport hydrophobic odorants through the aqueous fluid within olfactory sensilla to the underlying receptor proteins. The recent discovery of a large family of olfactory receptor genes in Drosophila raises new questions about the function, diversity, regulation, and evolution of the OBP family. We have investigated the OS-E and OS-F genes in a variety of Drosophila species. These studies highlight potential regions of functional significance in the OS-E and OS F proteins, which may include a region required for interaction with receptor proteins. Our results suggest that the two genes arose by an ancient gene duplication, and that in some lineages, one or the other gene has been lost. In D. virilis, the OS-F gene shows a different spatial pattern of expression than in D. melanogaster. One of the OS-F introns shows a striking degree of conservation between the two species, and we identify a putative regulatory sequence within this intron. Finally, a phylogenetic analysis places both OS-E and OS-F within a large family of insect OBPs and OBP-like proteins. PMID- 10790387 TI - Regulatory elements required for development of caenorhabditis elegans hermaphrodites are conserved in the tra-2 homologue of C. remanei, a male/female sister species. AB - The Caenorhabditis elegans hermaphrodite is essentially a female that produces sperm. In C. elegans, tra-2 promotes female fates and must be repressed to achieve hermaphrodite spermatogenesis. In an effort to learn how mating systems evolve, we have cloned tra-2 from C. remanei, the closest gonochoristic relative of C. elegans. We found its structure to be similar to that of Ce-tra-2 but its sequence to be divergent. RNA interference demonstrates that Cr-tra-2 promotes female fates. Two sites of tra-2 regulation are required for the onset of hermaphrodite spermatogenesis in C. elegans. One, the MX region of TRA-2, is as well conserved in C. remanei as it is in C. briggsae (another male/hermaphrodite species), suggesting that this control is not unique to hermaphrodites. Another, the DRE/TGE element of the tra-2 3' UTR, was not detected by sequence analysis. However, gel-shift assays demonstrate that a factor in C. remanei can bind specifically to the Cr-tra-2 3' UTR, suggesting that this translational control is also conserved. We propose that both controls are general and do not constitute a novel "switch" that enables sexual mosaicism in hermaphrodites. However, subtle quantitative or qualitative differences in their employment may underlie differences in mating system seen in Caenorhabditis. PMID- 10790390 TI - Genetic and molecular complexity of the position effect variegation modifier mod(mdg4) in Drosophila. AB - mod(mdg4), also known as E(var)3-93D, is involved in a variety of processes, such as gene silencing in position effect variegation (PEV), the control of gypsy insulator sequences, regulation of homeotic gene expression, and programmed cell death. We have isolated a large number of mod(mdg4) cDNAs, representing 21 different isoforms generated by alternative splicing. The deduced proteins are characterized by a common N terminus of 402 amino acids, including the BTB/POZ domain. Most of the variable C termini contain a new consensus sequence, including four positioned hydrophobic amino acids and a Cys(2)His(2) motif. Using specific antibodies for two protein isoforms, we demonstrate different distributions of the corresponding proteins on polytene chromosomes. Mutations in the genomic region encoding exons 1-4 show enhancement of PEV and homeotic transformation and affect viability and fertility. Homeotic and PEV phenotypes are enhanced by mutations in other trx-group genes. A transgene containing the common 5' region of mod(mdg4) that is present in all splice variants known so far partially rescues the recessive lethality of mod(mdg4) mutant alleles. Our data provide evidence that the molecular and genetic complexity of mod(mdg4) is caused by a large set of individual protein isoforms with specific functions in regulating the chromatin structure of different sets of genes throughout development. PMID- 10790389 TI - The Drosophila fl(2)d gene, required for female-specific splicing of Sxl and tra pre-mRNAs, encodes a novel nuclear protein with a HQ-rich domain. AB - The Drosophila gene female-lethal(2)d [fl(2)d] interacts genetically with the master regulatory gene for sex determination, Sex-lethal. Both genes are required for the activation of female-specific patterns of alternative splicing on transformer and Sex-lethal pre-mRNAs. We have used P-element-mediated mutagenesis to identify the fl(2)d gene. The fl(2)d transcription unit generates two alternatively spliced mRNAs that can encode two protein isoforms differing at their amino terminus. The larger isoform contains a domain rich in histidine and glutamine but has no significant homology to proteins in databases. Several lines of evidence indicate that this protein is responsible for fl(2)d function. First, the P-element insertion that inactivates fl(2)d interrupts this ORF. Second, amino acid changes within this ORF have been identified in fl(2)d mutants, and the nature of the changes correlates with the severity of the mutations. Third, all of the phenotypes associated with fl(2)d mutations can be rescued by expression of this cDNA in transgenic flies. Fl(2)d protein can be detected in extracts from Drosophila cell lines, embryos, larvae, and adult animals, without apparent differences between sexes, as well as in adult ovaries. Consistent with a possible function in posttranscriptional regulation, Fl(2)d protein has nuclear localization and is enriched in nuclear extracts. PMID- 10790391 TI - Drosophila wee1 has an essential role in the nuclear divisions of early embryogenesis. AB - In Drosophila, the maternally expressed mei-41 and grp genes are required for successful execution of the nuclear division cycles of early embryogenesis. In fission yeast, genes encoding similar kinases (rad3 and chk1, respectively) are components of a cell cycle checkpoint that delays mitosis by inhibitory phosphorylation of Cdk1. We have identified mutations in a gene encoding a Cdk1 inhibitory kinase, Drosophila wee1 (Dwee1). Like mei-41 and grp, Dwee1 is zygotically dispensable but is required maternally for completing the embryonic nuclear cycles. The arrest phenotype of Dwee1 mutants, as well as genetic interactions between Dwee1, grp, and mei-41 mutations, suggest that Dwee1 is functioning in the same regulatory pathway as these genes. These findings imply that inhibitory phosphorylation of Cdk1 by Dwee1 is required for proper regulation of the early syncytial cycles of embryogenesis. PMID- 10790392 TI - Offsetting effects of Wolbachia infection and heat shock on sperm production in Drosophila simulans: analyses of fecundity, fertility and accessory gland proteins. AB - Infection in Drosophila simulans with the endocellular symbiont Wolbachia pipientis results in egg lethality caused by failure to properly initiate diploid development (cytoplasmic incompatibility, CI). The relationship between Wolbachia infection and reproductive factors influencing male fitness has not been well examined. Here we compare infected and uninfected strains of D. simulans for (1) sperm production, (2) male fertility, and (3) the transfer and processing of two accessory gland proteins, Acp26Aa or Acp36De. Infected males produced significantly fewer sperm cysts than uninfected males over the first 10 days of adult life, and infected males, under varied mating conditions, had lower fertility compared to uninfected males. This fertility effect was due to neither differences between infected and uninfected males in the transfer and subsequent processing of accessory gland proteins by females nor to the presence of Wolbachia in mature sperm. We found that heat shock, which is known to decrease CI expression, increases sperm production to a greater extent in infected compared to uninfected males, suggesting a possible link between sperm production and heat shock. Given these results, the roles Wolbachia and heat shock play in mediating male gamete production may be important parameters for understanding the dynamics of infection in natural populations. PMID- 10790393 TI - Genetic analysis of a Y-chromosome region that induces triplosterile phenotypes and is essential for spermatid individualization in Drosophila melanogaster. AB - The heterochromatic Y chromosome of Drosophila melanogaster contains approximately 40 Mb of DNA but has only six loci mutable to male sterility. Region h1-h9 on YL, which carries the kl-3 and kl-5 loci, induces male sterility when present in three copies. We show that three separate segments within the region are responsible for the triplosterility and have an additive effect on male fertility. The triplosterile males displayed pleiotropic defects, beginning at early postmeiotic stages. However, the triplosterility was unaffected by kl-3 or kl-5 alleles. These data suggest that region h1-h9 is complex and may contain novel functions in addition to those of the previously identified kl-3 and kl-5 loci. The kl-3 and kl-5 mutations as well as deficiencies within region h1-h9 result in loss of the spermatid axonemal outer dynein arms. Examination using fluorescent probes showed that males deficient for h1-h3 or h4-h9 displayed a postmeiotic lesion with disrupted individualization complexes scattered along the spermatid bundle. In contrast, the kl-3 and kl-5 mutations had no effect on spermatid individualization despite the defect in the axonemes. These results demonstrate that region h1-h9 carries genetically separable functions: one required for spermatid individualization and the other essential for assembling the axonemal dynein arms. PMID- 10790394 TI - The Drosophila melanogaster sex determination gene sisA is required in yolk nuclei for midgut formation. AB - During sex determination, the sisterlessA (sisA) gene functions as one of four X:A numerator elements that set the alternative male or female regulatory states of the switch gene Sex-lethal. In somatic cells, sisA functions specifically in sex determination, but its expression pattern also hints at a role in the yolk cell, a syncytial structure believed to provide energy and nutrients to the developing embryo. Previous studies of sisA have been limited by the lack of a null allele, leaving open the possibility that sisA has additional functions. Here we report the isolation and molecular characterization of four new sisA alleles including two null mutations. Our findings highlight key aspects of sisA structure-function and reveal important qualitative differences between the effects of sisA and the other strong X:A numerator element, sisterlessB, on Sex lethal expression. We use genetic, expression, clonal, and phenotypic analyses to demonstrate that sisA has an essential function in the yolk nuclei of both sexes. In the absence of sisA, endoderm migration and midgut formation are blocked, suggesting that the yolk cell may have a direct role in larval gut development. To our knowledge, this is the first report of a requirement for the yolk nuclei in Drosophila development. PMID- 10790395 TI - Genetic modifiers of the Drosophila NSF mutant, comatose, include a temperature sensitive paralytic allele of the calcium channel alpha1-subunit gene, cacophony. AB - The N-ethylmaleimide-sensitive fusion protein (NSF) has been implicated in vesicle trafficking in perhaps all eukaryotic cells. The Drosophila comatose (comt) gene encodes an NSF homolog, dNSF1. Our previous work with temperature sensitive (TS) paralytic alleles of comt has revealed a function for dNSF1 at synapses, where it appears to prime synaptic vesicles for neurotransmitter release. To further examine the molecular basis of dNSF1 function and to broaden our analysis of synaptic transmission to other gene products, we have performed a genetic screen for mutations that interact with comt. Here we report the isolation and analysis of four mutations that modify TS paralysis in comt, including two intragenic modifiers (one enhancer and one suppressor) and two extragenic modifiers (both enhancers). The intragenic mutations will contribute to structure-function analysis of dNSF1 and the extragenic mutations identify gene products with related functions in synaptic transmission. Both extragenic enhancers result in TS behavioral phenotypes when separated from comt, and both map to loci not previously identified in screens for TS mutants. One of these mutations is a TS paralytic allele of the calcium channel alpha1-subunit gene, cacophony (cac). Analysis of synaptic function in these mutants alone and in combination will further define the in vivo functions and interactions of specific gene products in synaptic transmission. PMID- 10790397 TI - Genetic analysis of the hybrid male rescue locus of Drosophila. AB - Several hybrid rescue mutations-alleles that restore the viability of normally lethal hybrids-have been discovered in Drosophila melanogaster and its relatives. Here we analyze one of these genes, Hybrid male rescue (Hmr), asking two questions about its role in hybrid inviability. (1) Does the wild-type allele from D. melanogaster (Hmr(mel)) cause hybrid embryonic inviability? (2) Does Hmr(mel) cause hybrid larval inviability? Our results show that the wild-type product of Hmr is neither necessary nor sufficient for hybrid embryonic inviability. Hmr(mel) does, however, appear to lower the viability of hybrid larvae. The data further suggest (though do not prove) that Hmr(mel) acts as a gain-of-function poison in hybrids. These findings support previous claims that hybrid embryonic and larval lethalities are genetically distinct and suggest that Hmr(mel) is at least one of the proximate causes of hybrid larval inviability. PMID- 10790396 TI - Defective pigment granule biogenesis and aberrant behavior caused by mutations in the Drosophila AP-3beta adaptin gene ruby. AB - Lysosomal protein trafficking is a fundamental process conserved from yeast to humans. This conservation extends to lysosome-like organelles such as mammalian melanosomes and insect eye pigment granules. Recently, eye and coat color mutations in mouse (mocha and pearl) and Drosophila (garnet and carmine) were shown to affect subunits of the heterotetrameric adaptor protein complex AP-3 involved in vesicle trafficking. Here we demonstrate that the Drosophila eye color mutant ruby is defective in the AP-3beta subunit gene. ruby expression was found in retinal pigment and photoreceptor cells and in the developing central nervous system. ruby mutations lead to a decreased number and altered size of pigment granules in various cell types in and adjacent to the retina. Humans with lesions in the related AP-3betaA gene suffer from Hermansky-Pudlak syndrome, which is caused by defects in a number of lysosome-related organelles. Hermansky Pudlak patients have a reduced skin pigmentation and suffer from internal bleeding, pulmonary fibrosis, and visual system malfunction. The Drosophila AP 3beta adaptin also appears to be involved in processes other than eye pigment granule biogenesis because all ruby allele combinations tested exhibited defective behavior in a visual fixation paradigm. PMID- 10790399 TI - The mitochondrial genome of the brachiopod Laqueus rubellus. AB - The complete nucleotide sequence of the 14,017-bp mitochondrial (mt) genome of the articulate brachiopod Laqueus rubellus is presented. Being one of the smallest of known mt genomes, it has an extremely compact gene organization. While the same 13 polypeptides, two rRNAs, and 22 tRNAs are encoded as in most other animal mtDNAs, lengthy noncoding regions are absent, with the longest apparent intergenic sequence being 54 bp in length. Gene-end sequence overlaps are prevalent, and several stop codons are abbreviated. The genes are generally shorter, and three of the protein-coding genes are the shortest among known homologues. All of the tRNA genes indicate size reduction in either or both of the putative TPsiC and DHU arms compared with standard tRNAs. Possession of a TV (TPsiC arm-variable loop) replacement loop is inferred for tRNA(R) and tRNA(L tag). The DHU arm appears to be unpaired not only in tRNA(S-tct) and tRNA(S-tga), but also in tRNA(C), tRNA(I), and tRNA(T), a novel condition. All the genes are encoded in the same DNA strand, which has a base composition rich in thymine and guanine. The genome has an overall gene arrangement drastically different from that of any other organisms so far reported, but contains several short segments, composed of 2-3 genes, which are found in other mt genomes. Combined cooccurrence of such gene assortments indicates that the Laqueus mt genome is similar to the annelid Lumbricus, the mollusc Katharina, and the octocoral Sarcophyton mt genomes, each with statistical significance. Widely accepted schemes of metazoan phylogeny suggest that the similarity with the octocoral could have arisen through a process of convergent evolution, while it appears likely that the similarities with the annelid and the mollusc reflect phylogenetic relationships. PMID- 10790398 TI - A screen for modifiers of cyclin E function in Drosophila melanogaster identifies Cdk2 mutations, revealing the insignificance of putative phosphorylation sites in Cdk2. AB - In higher eukaryotes, cyclin E is thought to control the progression from G1 into S phase of the cell cycle by associating as a regulatory subunit with cdk2. To identify genes interacting with cyclin E, we have screened in Drosophila melanogaster for mutations that act as dominant modifiers of an eye phenotype caused by a Sevenless-CycE transgene that directs ectopic Cyclin E expression in postmitotic cells of eye imaginal disc and causes a rough eye phenotype in adult flies. The majority of the EMS-induced mutations that we have identified fall into four complementation groups corresponding to the genes split ends, dacapo, dE2F1, and Cdk2(Cdc2c). The Cdk2 mutations in combination with mutant Cdk2 transgenes have allowed us to address the regulatory significance of potential phosphorylation sites in Cdk2 (Thr 18 and Tyr 19). The corresponding sites in the closely related Cdk1 (Thr 14 and Tyr 15) are of crucial importance for regulation of the G2/M transition by myt1 and wee1 kinases and cdc25 phosphatases. In contrast, our results demonstrate that the equivalent sites in Cdk2 play no essential role. PMID- 10790401 TI - Evidence for recent invasion of the medaka fish genome by the Tol2 transposable element. AB - Tol2 is a transposable element of the terminal-inverted-repeat class, residing in the genome of the medaka fish Oryzias latipes. The genus Oryzias contains more than 10 species for which phylogenetic relationships have previously been estimated. To infer the history of Tol2 in this genus we performed genomic Southern blots and PCR analyses of 10 of the species. It was revealed that Tol2 occurs in 2 of the 10 species (O. curvinotus and O. latipes) and that the length and the restriction map structure of Tol2 are identical in the two cases. Further, sequencing analysis revealed an extremely low level of divergence compared with that in a nuclear gene. These results suggest recent incorporation of Tol2 into one or both of the two species, implying horizontal transfer of Tol2 from one species to the other or into them both from a common source. PMID- 10790400 TI - Analysis of chromosomal rearrangements induced by postmeiotic mutagenesis with ethylnitrosourea in zebrafish. AB - Mutations identified in zebrafish genetic screens allow the dissection of a wide array of problems in vertebrate biology. Most screens have examined mutations induced by treatment of spermatogonial (premeiotic) cells with the chemical mutagen N-ethyl-N-nitrosourea (ENU). Treatment of postmeiotic gametes with ENU induces specific-locus mutations at a higher rate than premeiotic regimens, suggesting that postmeiotic mutagenesis protocols could be useful in some screening strategies. Whereas there is extensive evidence that ENU induces point mutations in premeiotic cells, the range of mutations induced in postmeiotic zebrafish germ cells has been less thoroughly characterized. Here we report the identification and analysis of five mutations induced by postmeiotic ENU treatment. One mutation, snh(st1), is a translocation involving linkage group (LG) 11 and LG 14. The other four mutations, oep(st2), kny(st3), Df(LG 13)(st4), and cyc(st5), are deletions, ranging in size from less than 3 cM to greater than 20 cM. These results show that germ cell stage is an important determinant of the type of mutations induced. The induction of chromosomal rearrangements may account for the elevated frequency of specific-locus mutations observed after treatment of postmeiotic gametes with ENU. PMID- 10790402 TI - Heritability of the maternal meiotic drive system linked to Om and high resolution mapping of the Responder locus in mouse. AB - Matings between (C57BL/6 x DDK)F(1) females and C57BL/6 males result in a significant excess of offspring inheriting maternal DDK alleles in the central region of mouse chromosome 11 due to meiotic drive at the second meiotic division. We have shown previously that the locus subject to selection is in the vicinity of D11Mit66, a marker closely linked to the Om locus that controls the preimplantation embryo-lethal phenotype known as the "DDK syndrome." We have also shown that observation of meiotic drive in this system depends upon the genotype of the sire. Here we show that females that are heterozygous at Om retain the meiotic drive phenotype and define a 0.32-cM candidate interval for the Responder locus in this drive system. In addition, analysis of the inheritance of alleles at Om among the offspring of F(1) intercrosses indicates that the effect of the sire is determined by the sperm genotype at Om or a locus linked to Om. PMID- 10790403 TI - Genomic, transcriptional and mutational analysis of the mouse microphthalmia locus. AB - Mouse microphthalmia transcription factor (Mitf) mutations affect the development of four cell types: melanocytes, mast cells, osteoclasts, and pigmented epithelial cells of the eye. The mutations are phenotypically diverse and can be arranged in an allelic series. In humans, MITF mutations cause Waardenburg syndrome type 2A (WS2A) and Tietz syndrome, autosomal dominant disorders resulting in deafness and hypopigmentation. Mitf mice thus represent an important model system for the study of human disease. Here we report the complete exon/intron structure of the mouse Mitf gene and show it to be similar to the human gene. We also found that the mouse gene is transcriptionally complex and is capable of generating at least 13 different Mitf isoforms. Some of these isoforms are missing important functional domains of the protein, suggesting that they might play an inhibitory role in Mitf function and signal transduction. In addition, we determined the molecular basis for six microphthalmia mutations. Two of the mutations are reported for the first time here (Mitf(mi-enu198) and Mitf(mi-x39)), while the others (Mitf(mi-ws), Mitf(mi-bws), Mitf(mi-ew), and Mitf(mi-di)) have been described but the molecular basis for the mutation not determined. When analyzed in terms of the genomic and transcriptional data presented here, it is apparent that these mutations result from RNA processing or transcriptional defects. Interestingly, three of the mutations (Mitf(mi-x39), Mitf(mi-bws), and Mitf(mi-ws)) produce proteins that are missing important functional domains of the protein identified in in vitro studies, further confirming a biological role for these domains in the whole animal. PMID- 10790404 TI - Complete repopulation of mouse mitochondrial DNA-less cells with rat mitochondrial DNA restores mitochondrial translation but not mitochondrial respiratory function. AB - By the fusion of mtDNA-less (rho(0)) cells of Mus musculus domesticus with platelets from different species, mtDNA repopulated cybrids were obtained for finding the mtDNA species that could induce mitochondrial abnormalities. Expression of mitochondrial dysfunction might be expected in these cybrids due to incompatibility between nuclear and mitochondrial genomes from different species. The results showed that mouse rho(0) cells could receive mtDNA from a different mouse species, M. spretus, or even mtDNA from the rat, Rattus norvegicus, and that the introduced rat mtDNA, but not M. spretus mtDNA, caused mitochondrial dysfunction, even though rat mtDNA could restore normal mitochondrial translation in the cybrids. Considering that mitochondrial respiratory complexes consist of nuclear DNA- and mtDNA-coded polypeptides, these observations suggest that the nuclear and mitochondrial interactions required for replication, transcription, and translation of introduced rat mtDNA must be less stringently controlled than those required for formation of normal respiratory complexes. As no procedure for introduction of mutagenized mouse mtDNA into living cells has yet been established, these findings provide important insights into generating mtDNA knockout mice. PMID- 10790405 TI - Comparative genetics of nucleotide binding site-leucine rich repeat resistance gene homologues in the genomes of two dicotyledons: tomato and arabidopsis. AB - The presence of a single resistance (R) gene allele can determine plant disease resistance. The protein products of such genes may act as receptors that specifically interact with pathogen-derived factors. Most functionally defined R genes are of the nucleotide binding site-leucine rich repeat (NBS-LRR) supergene family and are present as large multigene families. The specificity of R-gene interactions together with the robustness of plant-pathogen interactions raises the question of their gene number and diversity in the genome. Genomic sequences from tomato showing significant homology to genes conferring race-specific resistance to pathogens were identified by systematically "scanning" the genome using a variety of primer pairs based on ubiquitous NBS motifs. Over 70 sequences were isolated and 10% are putative pseudogenes. Mapping of the amplified sequences on the tomato genetic map revealed their organization as mixed clusters of R-gene homologues that showed in many cases linkage to genetically characterized tomato resistance loci. Interspecific examination within Lycopersicon showed the existence of a null allele. Consideration of the tomato and potato comparative genetic maps unveiled conserved syntenic positions of R gene homologues. Phylogenetic clustering of R-gene homologues within tomato and other Solanaceae family members was observed but not with R-gene homologues from Arabidopsis thaliana. Our data indicate remarkably rapid evolution of R-gene homologues during diversification of plant families. PMID- 10790406 TI - The developmental expression of the maize regulatory gene Hopi determines germination-dependent anthocyanin accumulation. AB - The Hopi gene is a member of the maize r1 gene family. By genetic and molecular analyses we report that Hopi consists of a single gene residing on chromosome 10 approximately 4.5 cM distal to r1. Hopi conditions anthocyanin deposition in aleurone, scutellum, pericarp, root, mesocotyl, leaves, and anthers, thus representing one of the broadest specifications of pigmentation pattern reported to date of all the r1 genes. A unique feature of the Hopi gene is that seeds are completely devoid of pigment at maturity but show a photoinducible germination dependent anthocyanin accumulation in aleurone and scutellum. Our analysis has shown that the Hopi transcript is not present in scutellum of developing seeds but is induced only upon germination and that the simultaneous presence of both C1 and Hopi mRNAs is necessary to achieve A1 activation in scutella. We conclude that the expression pattern of the Hopi gene accounts for the germination dependent anthocyanin synthesis in scutella, whereas the developmental competence of germinating seeds to induce anthocyanin production in scutella results from the combination of the light-inducible expression of C1 and the developmentally regulated expression of the Hopi gene. PMID- 10790407 TI - Whole-genome characterization of embryonic stage inbreeding depression in a selfed loblolly pine family. AB - Inbreeding depression is important in the evolution of plant populations and mating systems. Previous studies have suggested that early-acting inbreeding depression in plants is primarily due to lethal alleles and possibly epistatic interactions. Recent advances in molecular markers now make genetic mapping a powerful tool to study the genetic architecture of inbreeding depression. We describe a genome-wide evaluation of embryonic viability loci in a selfed family of loblolly pine (Pinus taeda L.), using data from AFLP markers from an essentially complete genome map. Locus positions and effects were estimated from segregation ratios using a maximum-likelihood interval mapping procedure. We identified 19 loci showing moderately deleterious to lethal embryonic effects. These loci account for >13 lethal equivalents, greater than the average of 8.5 lethal equivalents reported for loblolly pine. Viability alleles show predominantly recessive action, although potential overdominance occurs at 3 loci. We found no evidence for epistasis in the distribution of pairwise marker correlations or in the regression of fitness on the number of markers linked to deleterious alleles. The predominant role of semilethal alleles in embryonic inbreeding depression has implications for the evolution of isolated populations and for genetic conservation and breeding programs in conifers. PMID- 10790408 TI - Analysis of extrachromosomal Ac/Ds transposable elements. AB - The mechanism of transposition of the maize Ac/Ds elements is not well understood. The true transposition intermediates are not known and it has not been possible to distinguish between excision models involving 8-bp staggered cuts or 1-bp staggered cuts followed by hairpin formation. In this work, we have analyzed extrachromosomal excision products to gain insight into the excision mechanism. Plasmid rescue was used to demonstrate that Ds excision is associated with the formation of circular molecules. In addition, we present evidence for the formation of linear extrachromosomal species during Ds excision. Sequences found at the termini of circular and linear elements showed a broad range of nucleotide additions or deletions, suggesting that these species are not true intermediates. Additional nucleotides adjacent to the termini in extrachromosomal elements were compared to the sequence of the original donor site. This analysis showed that: (1) the first nucleotide adjacent to the transposon end was significantly more similar to the first nucleotide flanking the element in the donor site than to a random sequence and (2) the second and farther nucleotides did not resemble the donor site. The implications of these findings for excision models are discussed. PMID- 10790409 TI - Highly recombinogenic regions at seed storage protein loci on chromosome 1DS of Aegilops tauschii, the D-genome donor of wheat. AB - A detailed RFLP map was constructed of the distal end of the short arm of chromosome 1D of Aegilops tauschii, the diploid D-genome donor species of hexaploid wheat. Ae. tauschii was used to overcome some of the limitations commonly associated with molecular studies of wheat such as low levels of DNA polymorphism. Detection of multiple loci by most RFLP probes suggests that gene duplication events have occurred throughout this chromosomal region. Large DNA fragments isolated from a BAC library of Ae. tauschii were used to determine the relationship between physical and genetic distance at seed storage protein loci located at the distal end of chromosome 1DS. Highly recombinogenic regions were identified where the ratio of physical to genetic distance was estimated to be <20 kb/cM. These results are discussed in relation to the genome-wide estimate of the relationship between physical and genetic distance. PMID- 10790410 TI - Spontaneous mutational effects on reproductive traits of arabidopsis thaliana. AB - A study of spontaneous mutation in Arabidopsis thaliana was initiated from a single inbred Columbia founder; 120 lines were established and advanced 17 generations by single-seed descent. Here, we report an assay of reproductive traits in a random set of 40 lines from generations 8 and 17, grown together at the same time with plants representing generation 0. For three reproductive traits, mean number of seeds per fruit, number of fruits, and dry mass of the infructescence, the means did not differ significantly among generations. Nevertheless, by generation 17, significant divergence among lines was detected for each trait, indicating accumulation of mutations in some lines. Standardized measures of mutational variance accord with those obtained for other organisms. These findings suggest that the distribution of mutational effects for these traits is approximately symmetric, in contrast to the usual assumption that mutations have predominantly negative effects on traits directly related to fitness. Because distinct generations were grown contemporaneously, each line was represented by three sublines, and seeds were equal in age, these estimates are free of potentially substantial sources of bias. The finding of an approximately symmetric distribution of mutational effects invalidates the standard approach for inferring properties of spontaneous mutation and necessitates further development of more general approaches that avoid restrictions on the distribution of mutational effects. PMID- 10790411 TI - Two independent loci control agamospermy (Apomixis) in the triploid flowering plant Erigeron annuus. AB - Asexual seed production (agamospermy) via gametophytic apomixis in flowering plants typically involves the formation of an unreduced megagametophyte (via apospory or diplospory) and the parthenogenetic development of the unreduced egg cell into an embryo. Agamospermy is almost exclusively restricted to polyploids. In this study, the genetic basis of agamospermy was investigated in a segregating population of 130 F(1)'s from a cross between triploid (2n = 27) agamospermous Erigeron annuus and sexual diploid (2n = 18) E. strigosus. Correlations between markers and phenotypes and linkage analysis were performed on 387 segregating amplified fragment length polymorphisms (AFLPs). Results show that four closely linked markers with polysomic inheritance are significantly associated with parthenogenesis and that 11 cosegregating markers with univalent inheritance are completely associated with diplospory. This indicates that diplospory and parthenogenesis are unlinked and inherited independently. Further, the absence of agamospermy in diploid F(1)'s appears to be best explained by a combination of recessive-lethal gametophytic selection against the parthenogenetic locus and univalent inheritance of the region bearing diplospory. These results may have major implications for attempts to manipulate agamospermy for agricultural purposes and for interpreting the evolution of the trait. PMID- 10790412 TI - Quantitative trait loci mapping in F(2) crosses between outbred lines. AB - We develop a mixed-model approach for QTL analysis in crosses between outbred lines that allows for QTL segregation within lines as well as for differences in mean QTL effects between lines. We also propose a method called "segment mapping" that is based in partitioning the genome in a series of segments. The expected change in mean according to percentage of breed origin, together with the genetic variance associated with each segment, is estimated using maximum likelihood. The method also allows the estimation of differences in additive variances between the parental lines. Completely fixed random and mixed models together with segment mapping are compared via simulation. The segment mapping and mixed-model behaviors are similar to those of classical methods, either the fixed or random models, under simple genetic models (a single QTL with alternative alleles fixed in each line), whereas they provide less biased estimates and have higher power than fixed or random models in more complex situations, i.e., when the QTL are segregating within the parental lines. The segment mapping approach is particularly useful to determining which chromosome regions are likely to contain QTL when these are linked. PMID- 10790413 TI - Selective mapping: a strategy for optimizing the construction of high-density linkage maps. AB - Historically, linkage mapping populations have consisted of large, randomly selected samples of progeny from a given pedigree or cell lines from a panel of radiation hybrids. We demonstrate that, to construct a map with high genome-wide marker density, it is neither necessary nor desirable to genotype all markers in every individual of a large mapping population. Instead, a reduced sample of individuals bearing complementary recombinational or radiation-induced breakpoints may be selected for genotyping subsequent markers from a large, but sparsely genotyped, mapping population. Choosing such a sample can be reduced to a discrete stochastic optimization problem for which the goal is a sample with breakpoints spaced evenly throughout the genome. We have developed several different methods for selecting such samples and have evaluated their performance on simulated and actual mapping populations, including the Lister and Dean Arabidopsis thaliana recombinant inbred population and the GeneBridge 4 human radiation hybrid panel. Our methods quickly and consistently find much-reduced samples with map resolution approaching that of the larger populations from which they are derived. This approach, which we have termed selective mapping, can facilitate the production of high-quality, high-density genome-wide linkage maps. PMID- 10790414 TI - Fine mapping of quantitative trait loci using linkage disequilibria with closely linked marker loci. AB - A multimarker linkage disequilibrium mapping method was developed for the fine mapping of quantitative trait loci (QTL) using a dense marker map. The method compares the expected covariances between haplotype effects given a postulated QTL position to the covariances that are found in the data. The expected covariances between the haplotype effects are proportional to the probability that the QTL position is identical by descent (IBD) given the marker haplotype information, which is calculated using the genedropping method. Simulation results showed that a QTL was correctly positioned within a region of 3, 1.5, or 0.75 cM in 70, 62, and 68%, respectively, of the replicates using markers spaced at intervals of 1, 0.5, and 0.25 cM, respectively. These results were rather insensitive to the number of generations since the QTL occurred and to the effective population size, except that 10 generations yielded rather poor estimates of the QTL position. The position estimates of this multimarker disequilibrium mapping method were more accurate than those from a single marker transmission disequilibrium test. A general approach for identifying QTL is suggested, where several stages of disequilibrium mapping are used with increasingly dense marker spacing. PMID- 10790415 TI - Codon-substitution models for heterogeneous selection pressure at amino acid sites. AB - Comparison of relative fixation rates of synonymous (silent) and nonsynonymous (amino acid-altering) mutations provides a means for understanding the mechanisms of molecular sequence evolution. The nonsynonymous/synonymous rate ratio (omega = d(N)d(S)) is an important indicator of selective pressure at the protein level, with omega = 1 meaning neutral mutations, omega < 1 purifying selection, and omega > 1 diversifying positive selection. Amino acid sites in a protein are expected to be under different selective pressures and have different underlying omega ratios. We develop models that account for heterogeneous omega ratios among amino acid sites and apply them to phylogenetic analyses of protein-coding DNA sequences. These models are useful for testing for adaptive molecular evolution and identifying amino acid sites under diversifying selection. Ten data sets of genes from nuclear, mitochondrial, and viral genomes are analyzed to estimate the distributions of omega among sites. In all data sets analyzed, the selective pressure indicated by the omega ratio is found to be highly heterogeneous among sites. Previously unsuspected Darwinian selection is detected in several genes in which the average omega ratio across sites is <1, but in which some sites are clearly under diversifying selection with omega > 1. Genes undergoing positive selection include the beta-globin gene from vertebrates, mitochondrial protein coding genes from hominoids, the hemagglutinin (HA) gene from human influenza virus A, and HIV-1 env, vif, and pol genes. Tests for the presence of positively selected sites and their subsequent identification appear quite robust to the specific distributional form assumed for omega and can be achieved using any of several models we implement. However, we encountered difficulties in estimating the precise distribution of omega among sites from real data sets. PMID- 10790416 TI - The coalescent with gene conversion. AB - In this article we develop a coalescent model with intralocus gene conversion. The distribution of the tract length is geometric in concordance with results published in the literature. We derive a simulation scheme and deduce a number of analytical results for this coalescent with gene conversion. We compare patterns of variability in samples simulated according to the coalescent with recombination with similar patterns simulated according to the coalescent with gene conversion alone. Further, an expression for the expected number of topology shifts in a sample of present-day sequences caused by gene conversion events is derived. PMID- 10790417 TI - Quantitative trait loci: a meta-analysis. AB - This article presents a method to combine QTL results from different independent analyses. This method provides a modified Akaike criterion that can be used to decide how many QTL are actually represented by the QTL detected in different experiments. This criterion is computed to choose between models with one, two, three, etc., QTL. Simulations are carried out to investigate the quality of the model obtained with this method in various situations. It appears that the method allows the length of the confidence interval of QTL location to be consistently reduced when there are only very few "actual" QTL locations. An application of the method is given using data from the maize database available online at http://www. agron.missouri.edu/. PMID- 10790418 TI - The Mre11p/Rad50p/Xrs2p complex and the Tel1p function in a single pathway for telomere maintenance in yeast. AB - The Mre11p/Rad50p/Xrs2p complex is involved in the repair of double-strand DNA breaks, nonhomologous end joining, and telomere length regulation. TEL1 is primarily involved in telomere length regulation. By an epistasis analysis, we conclude that Tel1p and the Mre11p/Rad50p/Xrs2p complex function in a single pathway of telomere length regulation. PMID- 10790419 TI - Modulating phagocyte activation: the pros and cons of Helicobacter pylori virulence factors. PMID- 10790420 TI - T cells and the regulation of herpes simplex virus latency and reactivation. PMID- 10790421 TI - CD8(+) T cells can block herpes simplex virus type 1 (HSV-1) reactivation from latency in sensory neurons. AB - Recurrent herpes simplex virus type 1 (HSV-1) disease usually results from reactivation of latent virus in sensory neurons and transmission to peripheral sites. Therefore, defining the mechanisms that maintain HSV-1 in a latent state in sensory neurons may provide new approaches to reducing susceptibility to recurrent herpetic disease. After primary HSV-1 corneal infection, CD8(+) T cells infiltrate the trigeminal ganglia (TGs) of mice, and are retained in latently infected ganglia. Here we demonstrate that CD8(+) T cells that are present in the TGs at the time of excision can maintain HSV-1 in a latent state in sensory neurons in ex vivo TG cultures. Latently infected neurons expressed viral genome and some expressed HSV-1 immediate early and early proteins, but did not produce HSV-1 late proteins or infectious virions. Addition of anti-CD8alpha monoclonal antibody 5 d after culture initiation induced HSV-1 reactivation, as demonstrated by production of viral late proteins and infectious virions. Thus, CD8(+) T cells can prevent HSV-1 reactivation without destroying the infected neurons. We propose that when the intrinsic capacity of neurons to inhibit HSV-1 reactivation from latency is compromised, production of HSV-1 immediate early and early proteins might activate CD8(+) T cells aborting virion production. PMID- 10790423 TI - Alymphoplasia (aly)-type nuclear factor kappaB-inducing kinase (NIK) causes defects in secondary lymphoid tissue chemokine receptor signaling and homing of peritoneal cells to the gut-associated lymphatic tissue system. AB - Alymphoplasia (aly) mice, which carry a point mutation in the nuclear factor kappaB-inducing kinase (NIK) gene, are characterized by the systemic absence of lymph nodes and Peyer's patches, disorganized splenic and thymic architectures, and immunodeficiency. Another unique feature of aly/aly mice is that their peritoneal cavity contains more B1 cells than normal and aly/+ mice. Transfer experiments of peritoneal lymphocytes from aly/aly mice into recombination activating gene (RAG)-2(-/-) mice revealed that B and T cells fail to migrate to other lymphoid tissues, particularly to the gut-associated lymphatic tissue system. In vivo homing defects of aly/aly peritoneal cells correlated with reduction of their in vitro chemotactic responses to secondary lymphoid tissue chemokine (SLC) and B lymphocyte chemoattractant (BLC). The migration defect of aly/aly lymphocytes was not due to a lack of expression of chemokines and their receptors, but rather to impaired signal transduction downstream of the receptors for SLC, indicating that NIK is involved in the chemokine signaling pathway known to couple only with G proteins. The results showed that the reduced serum levels of immunoglobulins (Igs) and the absence of class switch to IgA in aly/aly mice are due, at least in part, to a migration defect of lymphocytes to the proper microenvironment where B cells proliferate and differentiate into Ig-producing cells. PMID- 10790422 TI - The neutrophil-activating protein (HP-NAP) of Helicobacter pylori is a protective antigen and a major virulence factor. AB - Helicobacter pylori infection induces the appearance of inflammatory infiltrates, consisting mainly of neutrophils and monocytes, in the human gastric mucosa. A bacterial protein with neutrophil activating activity (HP-NAP) has been previously identified, but its role in infection and immune response is still largely unknown. Here, we show that vaccination of mice with HP-NAP induces protection against H. pylori challenge, and that the majority of infected patients produce antibodies specific for HP-NAP, suggesting an important role of this factor in immunity. We also show that HP-NAP is chemotactic for human leukocytes and that it activates their NADPH oxidase to produce reactive oxygen intermediates, as demonstrated by the translocation of its cytosolic subunits to the plasma membrane, and by the lack of activity on chronic granulomatous disease leukocytes. This stimulating effect is strongly potentiated by tumor necrosis factor alpha and interferon gamma and is mediated by a rapid increase of the cytosolic calcium concentration. The activation of leukocytes induced by HP-NAP is completely inhibited by pertussis toxin, wortmannin, and PP1. On the basis of these results, we conclude that HP-NAP is a virulence factor important for the H. pylori pathogenic effects at the site of infection and a candidate antigen for vaccine development. PMID- 10790424 TI - M11L: a novel mitochondria-localized protein of myxoma virus that blocks apoptosis of infected leukocytes. AB - M11L, a novel 166-amino acid membrane-associated protein expressed by the poxvirus, myxoma virus, was previously found to modulate apoptosis after infection of rabbit leukocytes. Furthermore, infection of rabbits with an M11L knockout virus unexpectedly produced lesions with a profound proinflammatory phenotype. We show here that M11L is antiapoptotic when expressed independently of other viral proteins, and is directed specifically to mitochondria by a short COOH-terminal region that is necessary and sufficient for targeting. This targeting region consists of a hydrophobic domain flanked by basic amino acid residues, adjacent to a positively charged tail. M11L blocks staurosporine induced apoptosis by preventing mitochondria from undergoing a permeability transition, and the mitochondrial localization of this protein is essential for this function. We show that M11L is specifically required to inhibit the apoptotic response of monocytes/macrophages during virus infection, as cells of this lineage undergo apoptosis when infected with the M11L knockout virus. As monocyte apoptosis is uniquely proinflammatory, we propose that this observation reconciles the paradoxical proapoptotic and proinflammatory phenotypes of the M11L knockout virus. We suggest that apoptosis of tissue macrophages represents an important antiviral defense, and that the inhibition of apoptosis by viral proteins can be directed in a cell-specific fashion. PMID- 10790425 TI - Analysis of successful immune responses in persons infected with hepatitis C virus. AB - Although hepatitis C virus (HCV) infection is very common, identification of patients during acute infection is rare. Consequently, little is known about the immune response during this critical stage of the disease. We analyzed the T lymphocyte response during and after acute resolving HCV infection in three persons, using interferon (IFN)-gamma enzyme-linked immunospot (ELISPOT) and human histocompatibility leukocyte antigen (HLA) peptide tetramer assays. Acute infection was associated with a broadly directed T helper and cytotoxic T lymphocyte (CTL) response, which persisted after resolution of clinical hepatitis and clearance of viremia. At the earliest time point studied, highly activated CTL populations were observed that temporarily failed to secrete IFN-gamma, a "stunned" phenotype, from which they recovered as viremia declined. In long-term HCV-seropositive persons, CTL responses were more common in persons who had cleared viremia compared with those with persistent viremia, although the frequencies of HCV-specific CTLs were lower than those found in persons during and after resolution of acute HCV infection. These studies demonstrate a strong and persistent CTL response in resolving acute HCV infection, and provide rationale to explore immune augmentation as a therapeutic intervention in chronic HCV infection. PMID- 10790426 TI - Cytoplasmic processing is a prerequisite for presentation of an endogenous antigen by major histocompatibility complex class II proteins. AB - Biochemical and functional studies have demonstrated major histocompatibility complex (MHC) class II-restricted presentation of select epitopes derived from cytoplasmic antigens, with few insights into the processing reactions necessary for this alternate pathway. Efficient presentation of an immunodominant epitope derived from glutamate decarboxylase (GAD) was observed regardless of whether this antigen was delivered exogenously or via a cytoplasmic route into human histocompatibility leukocyte antigen class II-DR4(+) antigen-presenting cells. Presentation of exogenous as well as cytoplasmic GAD required the intersection of GAD peptides and newly synthesized class II proteins. By contrast, proteolytic processing of this antigen was highly dependent upon the route of antigen delivery. Exogenous GAD followed the classical pathway for antigen processing, with an absolute requirement for endosomal/lysosomal acidification as well as cysteine and aspartyl proteases resident within these organelles. Presentation of endogenous GAD was dependent upon the action of cytoplasmic proteases, including the proteasome and calpain. Thus, translocation of processed antigen from the cytoplasm into membrane organelles is necessary for class II-restricted presentation via this alternate pathway. Further trimming of these peptides after translocation was mediated by acidic proteases within endosomes/lysosomes, possibly after or before class II antigen binding. These studies suggest that processing of exogenous and cytoplasmic proteins occurs through divergent but overlapping pathways. Furthermore, two cytoplasmic proteases, the proteasome and calpain, appear to play important roles in MHC class II-restricted antigen presentation. PMID- 10790427 TI - Degradation of transcription factor RFX5 during the inhibition of both constitutive and interferon gamma-inducible major histocompatibility complex class I expression in chlamydia-infected cells. AB - We have previously shown that the obligate intracellular pathogen chlamydia can suppress interferon (IFN)-gamma-inducible major histocompatibility complex (MHC) class II expression in infected cells by degrading upstream stimulation factor (USF)-1. We now report that chlamydia can also inhibit both constitutive and IFN gamma-inducible MHC class I expression in the infected cells. The inhibition of MHC class I molecule expression correlates well with degradation of RFX5, an essential downstream transcription factor required for both the constitutive and IFN-gamma-inducible MHC class I expression. We further demonstrate that a lactacystin-sensitive proteasome-like activity identified in chlamydia-infected cell cytosolic fraction can degrade both USF-1 and RFX5. This proteasome-like activity is dependent on chlamydial but not host protein synthesis. Host preexisting proteasomes may not be required for the unique proteasome-like activity. These observations suggest that chlamydia-secreted factors may directly participate in the proteasome-like activity. Efforts to identify the chlamydial factors are underway. These findings provide novel information on the molecular mechanisms of chlamydial evasion of host immune recognition. PMID- 10790429 TI - Differential regulation of B cell development, activation, and death by the src homology 2 domain-containing 5' inositol phosphatase (SHIP). AB - Although the Src homology 2 domain-containing 5' inositol phosphatase (SHIP) is a well-known mediator of inhibitory signals after B cell antigen receptor (BCR) coaggregation with the low affinity Fc receptor, it is not known whether SHIP functions to inhibit signals after stimulation through the BCR alone. Here, we show using gene-ablated mice that SHIP is a crucial regulator of BCR-mediated signaling, B cell activation, and B cell development. We demonstrate a critical role for SHIP in termination of phosphatidylinositol 3,4,5-triphosphate (PI[3,4,5]P(3)) signals that follow BCR aggregation. Consistent with enhanced PI(3,4,5)P(3) signaling, we find that splenic B cells from SHIP-deficient mice display enhanced sensitivity to BCR-mediated induction of the activation markers CD86 and CD69. We further demonstrate that SHIP regulates the rate of B cell development in the bone marrow and spleen, as B cell precursors from SHIP deficient mice progress more rapidly through the immature and transitional developmental stages. Finally, we observe that SHIP-deficient B cells have increased resistance to BCR-mediated cell death. These results demonstrate a central role for SHIP in regulation of BCR signaling and B cell biology, from signal driven development in the bone marrow and spleen, to activation and death in the periphery. PMID- 10790428 TI - The serpin proteinase inhibitor 9 is an endogenous inhibitor of interleukin 1beta converting enzyme (caspase-1) activity in human vascular smooth muscle cells. AB - Interleukin-1beta-converting enzyme (ICE, caspase-1) regulates key steps in inflammation and immunity, by activating the proinflammatory cytokines interleukin (IL-)1beta and IL-18, or mediating apoptotic processes. We recently provided evidence for the regulation of caspase-1 activity via an endogenous inhibitor expressed by human vascular smooth muscle cells (SMCs) (Schonbeck, U., M. Herzberg, A. Petersen, C. Wohlenberg, J. Gerdes, H.-D. Flad, and H. Loppnow. 1997. J. Exp. Med. 185:1287-1294). However, the molecular identity of this endogenous inhibitor remained undefined. We report here that the serine proteinase inhibitor (serpin) PI-9 accounts for the endogenous caspase-1 inhibitory activity in human SMCs and prevents processing of the enzyme's natural substrates, IL-1beta and IL-18 precursor. Treatment of SMC lysates with anti-PI-9 antibody abrogated the caspase-1 inhibitory activity and coprecipitated the enzyme, demonstrating protein-protein interaction. Furthermore, PI-9 antisense oligonucleotides coordinately reduced PI-9 expression and promoted IL-1beta release. Since SMCs comprise the majority of cells in the vascular wall, and because IL-1 is implicated in atherogenesis, we tested the biological validity of our in vitro findings within human atheroma in situ. The unaffected arterial wall contains abundant and homogeneously distributed PI-9. In human atherosclerotic lesions, however, PI-9 expression correlated inversely with immunoreactive IL 1beta, supporting a potential role of the endogenous caspase-1 inhibitor in this chronic inflammatory disease. Thus, our results provide new insights into the regulation of this enzyme involved in immune and inflammatory processes of chronic inflammatory diseases, and point to an endogenous antiinflammatory action of PI-9, dysregulated in a prevalent human disease. PMID- 10790430 TI - Molecular basis for leukocyte integrin alpha(E)beta(7) adhesion to epithelial (E) cadherin. AB - Cadherins are expressed in tissue-restricted patterns and typically mediate homophilic adhesion. Cadherins also mediate lymphocyte adhesion, providing the opportunity for lymphocyte attachment to parenchymal cells. The best characterized example of lymphocyte adhesion to a tissue-specific cell adhesion molecule, as opposed to a vascular endothelial adhesion molecule, is the interaction between integrin alpha(E)beta(7) on intraepithelial lymphocytes and E cadherin on epithelial cells. However, the molecular basis for an integrin cadherin interaction is not well defined. Realization that the cadherin domain adopts a topology similar to the immunoglobulin (Ig) fold suggested that integrin recognition of E-cadherin might be similar to recognition of Ig superfamily ligands. Thus, we modeled domain 1 of human E-cadherin and studied the role of solvent-exposed loops that connect Ig-like core-forming beta strands. Mutational analyses localized the integrin alpha(E)beta(7) recognition site to the top of domain 1 at the face formed by the BC and FG loops, a site distinct from the region recognized in intercellular adhesion molecule (ICAM)-1, -2, and -3, mucosal addressin cell adhesion molecule 1 (MAdCAM-1), vascular cell adhesion molecule 1 (VCAM-1), and fibronectin by their integrin ligands. Moreover, the integrin alpha(E)beta(7) binding site is distinct from the homophilic binding site on E-cadherin. These studies provide a conceptual basis for integrin cadherin binding and extend the model that an Ig-like fold can serve as a scaffold for recognition. PMID- 10790431 TI - Enterocyte expression of interleukin 7 induces development of gammadelta T cells and Peyer's patches. AB - The intestinal mucosa is suggested to support extrathymic T cell development, particularly for T cell receptor (TCR)-gammadelta intraepithelial lymphocytes (IELs). TCR-gammadelta cell development requires interleukin (IL)-7; IL-7(-/)- or IL-7 receptor(-/)- mice lack TCR-gammadelta cells. Using the intestinal fatty acid binding protein (iFABP) promoter, we reinstated expression of IL-7 to mature enterocytes of IL-7(-/)- mice (iFABP-IL7). In iFABP-IL7 mice, TCR-gammadelta IELs were restored, as were cryptopatches and Peyer's patches. TCR-gammadelta cells remained absent from all other tissues. Likewise, T cell development in thymus and B cell maturation in the bone marrow and spleen retained the IL-7(-/)- phenotype. Thus, IL-7 expression by enterocytes was sufficient for extrathymic development of TCR-gammadelta cells in situ within the intestinal epithelium and was crucial for organization of mucosal lymphoid tissue. PMID- 10790432 TI - Inhibition of CD83 cell surface expression during dendritic cell maturation by interference with nuclear export of CD83 mRNA. AB - Dendritic cells (DCs), nature's adjuvant, must mature to sensitize T cells. However, although the maturation process is essential, it is not yet fully understood at the molecular level. In this study, we investigated the course of expression of the unique hypusine-containing protein eukaryotic initiation factor 5A (eIF-5A), which is part of a particular RNA nuclear export pathway, during in vitro generation of human DCs. We show that eIF-5A expression is significantly upregulated during DC maturation. Furthermore, an inhibitor of the hypusine modification, GC7 (N(1)-guanyl-1, 7-diaminoheptane), prevents CD83 surface expression by apparently interfering with nucleocytoplasmic translocation of the CD83 mRNA and, importantly, significantly inhibits DC-mediated T lymphocyte activation. The data presented suggest that CD83 mRNA is transported from the nucleus to the cytoplasm via a specific nuclear export pathway and that hypusine formation appears to be essential for the maturation of functional DCs. Therefore, pharmacological interference with hypusine formation may provide a new possibility to modulate DC function. PMID- 10790433 TI - Phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG), a novel ubiquitously expressed transmembrane adaptor protein, binds the protein tyrosine kinase csk and is involved in regulation of T cell activation. AB - According to a recently proposed hypothesis, initiation of signal transduction via immunoreceptors depends on interactions of the engaged immunoreceptor with glycosphingolipid-enriched membrane microdomains (GEMs). In this study, we describe a novel GEM-associated transmembrane adaptor protein, termed phosphoprotein associated with GEMs (PAG). PAG comprises a short extracellular domain of 16 amino acids and a 397-amino acid cytoplasmic tail containing ten tyrosine residues that are likely phosphorylated by Src family kinases. In lymphoid cell lines and in resting peripheral blood alpha/beta T cells, PAG is expressed as a constitutively tyrosine-phosphorylated protein and binds the major negative regulator of Src kinases, the tyrosine kinase Csk. After activation of peripheral blood alpha/beta T cells, PAG becomes rapidly dephosphorylated and dissociates from Csk. Expression of PAG in COS cells results in recruitment of endogenous Csk, altered Src kinase activity, and impaired phosphorylation of Src specific substrates. Moreover, overexpression of PAG in Jurkat cells downregulates T cell receptor-mediated activation of the transcription factor nuclear factor of activated T cells. These findings collectively suggest that in the absence of external stimuli, the PAG-Csk complex transmits negative regulatory signals and thus may help to keep resting T cells in a quiescent state. PMID- 10790434 TI - Primary role for Gi protein signaling in the regulation of interleukin 12 production and the induction of T helper cell type 1 responses. AB - We explored the role of Gi protein signaling in the regulation of interleukin (IL)-12 production and T helper cell type 1 (Th1) T cell differentiation. In initial studies, we showed that treatment of normal mice with pertussis toxin (PT), which inhibits Gi protein signaling, enhanced the capacity of splenocytes to produce IL-12 in response to both microbial and nonmicrobial stimuli. In addition, PT treatment increased the production of tumor necrosis factor (TNF) alpha and IL-10 by stimulated cells. These findings were corroborated by the fact that untreated Gi2alpha(2/-) mice exhibited enhanced production of IL-12 and TNF alpha by splenocytes, and of IL-12 p40 by purified spleen CD8alpha(+) lymphoid dendritic cells. Finally, we showed that while normal BALB/c mice infected with Leishmania major exhibited a nonhealing phenotype, those treated with PT when infection was initiated exhibited a healing phenotype along with an enhancement of leishmania-specific Th1 responses in draining lymph nodes. Further, healing was prevented by coadministration of anti-IL-12 and PT. These data demonstrate that endogenous Gi protein signaling has a primary role in the regulation of IL 12 production and the induction of Th1 responses in vivo. PMID- 10790435 TI - Induction and suppression of collagen-induced arthritis is dependent on distinct fcgamma receptors. AB - Receptors for immunoglobulin (Ig)G (FcgammaRs) are important for the antibody mediated effector functions of the immune system. FcgammaRI and FcgammaRIII trigger cell activation through a common gamma chain, whereas FcgammaRII acts as a negative regulator of antibody production and immune complex-triggered activation. Here we describe the in vivo consequences of FcgammaR deficiency in a mouse model of human rheumatoid arthritis. FcRgamma chain-deficient mice on arthritis-susceptible DBA/1 background were immunized with collagen for induction of collagen-induced arthritis. The DBA/1 mice lacking FcRgamma chain were protected from collagen-induced arthritis in contrast to wild-type mice, although both groups produced similar levels of IgG anticollagen antibodies. In comparison, DBA/1 mice lacking FcgammaRII developed an augmented IgG anticollagen response and arthritis. These observations suggest a crucial role of FcgammaRI and FcgammaRIII in triggering autoimmune arthritis. PMID- 10790437 TI - Scientific rationale for the use of antenatal glucocorticoids to promote fetal development. PMID- 10790436 TI - A processed pseudogene codes for a new antigen recognized by a CD8(+) T cell clone on melanoma. AB - The M88.7 T cell clone recognizes an antigen presented by HLA B*1302 on the melanoma cell line M88. A cDNA encoding this antigen (NA88-A) was isolated using a library transfection approach. Analysis of the genomic gene's sequence identified it is a processed pseudogene, derived from a retrotranscript of mRNA coding for homeoprotein HPX42B. The NA88-A gene exhibits several premature stop codons, deletions, and insertions relative to the HPX42B gene. In NA88-A RNA, a short open reading frame codes for the peptide MTQGQHFLQKV from which antigenic peptides are derived; a stop codon follows the peptide's COOH-terminal Val codon. Part of the HPX42B mRNA's 3' untranslated region codes for a peptide of similar sequence (MTQGQHFSQKV). If produced, this peptide can be recognized by M88.7 T cells. However, in HPX42B mRNA, the peptide's COOH-terminal Val codon is followed by a Trp codon. As a result, expression of HPX42B mRNA does not lead to antigen production. A model is proposed for events that participated in creation of a gene coding for a melanoma antigen from a pseudogene. PMID- 10790438 TI - Clinical use of antenatal corticosteroids: benefits and risks. PMID- 10790439 TI - The Gold Medal Prize in the Conjoint M.Orth. exam of the Royal College of Surgeons of Edinburgh held in Hong Kong, 1996. PMID- 10790440 TI - How much space is created from expansion or premolar extraction? AB - The aim of this study was first to investigate the relationship between maxillary arch expansion and change in arch depth (overjet), and secondly to quantify the reduction in maxillary arch depth following extraction of 4?4 with complete space closure. A model of maxillary typodont teeth was constructed to allow expansion and premolar removal. Arch dimensions were recorded using a reflex microscope. A linear relationship was found between arch expansion and reduction of the arch depth. When the premolars were removed, there was a greater reduction in arch depth than the mesio-distal width of these teeth. PMID- 10790441 TI - Skeletal muscle function and fibre types: the relationship between occlusal function and the phenotype of jaw-closing muscles in human. AB - Mammalian skeletal muscle cells are composed of repeated sarcomeric units containing thick and thin filaments of myosin and actin, respectively. Excitation of the myosin ATPase enzyme is possible only with presence of Mg-ATP and Ca(2+). Skeletal muscle fibres may be classified into several types according to the isoform of myosin they contain. Nine isoforms of myosin heavy chain are known to exist in mammalian skeletal muscle including type I, IIA, IIB, IIX, IIM, alpha, neonatal, embryonic, and extra-ocular. Healthy adult human limb skeletal muscle contains type I, IIA, IIB, and IIX myosin heavy chains. The jaw-closing muscles of most carnivores and primates have tissue-specific expression of the type IIM or 'type II masticatory' myosin heavy chain. Adult human jaw-closing muscles, however, do not contain IIM myosin. Rather, they express type I, IIA, IIX (as in human limb muscle), and myosins typically expressed in developing or cardiac muscle. The morphology of human jaw-closing muscle fibres is also unusual in that the type II fibres are of smaller diameter that type I fibres, except in cases of increased function and hypertrophy. This paper describes the relationship of fibre types and motor unit function to changes in human occlusion and masticatory activity. Refereed Scientific Paper PMID- 10790442 TI - Three-dimensional facial growth studied by optical surface scanning. AB - The objective of the investigation was to study the three-dimensional growth of the face, and to examine the hypothesis that there are three-dimensional differences between the faces of boys and girls. The subjects comprised 132 British Caucasians aged 5-10 years measured by optical surface scanning in this cross-sectional study. Average scans for each age and sex subgroup were superimposed to assess the differences with age and sex. Males were generally larger than females. The greatest difference was between the facial heights and the least in the mid-facial dimensions. The face height of both sexes increased by an average of 3-4 mm annually. Mid-face prominence and width altered little. Mandibular width increased by 1-3 mm a year, rising to 3-5 mm in some years at the inferior areas of the mandibular region. Mandibular prominence also increased. Nose height and prominence and alar base width increased by 2 mm per year on average. Dorsum width changed little. Boys were generally larger than girls. Growth in facial height was greatest. Mid-face prominence and width changed little with age, whilst the prominence and width of the lower face increased more. Nasal prominence and alar base width increased at most ages. Dimensions changed more than reported by cephalometric studies, possibly as this study included the soft tissues. Refereed Scientific Paper PMID- 10790443 TI - An evaluation of the stresses generated in a bonded orthodontic attachment by three different load cases using the Finite Element Method of stress analysis. AB - The objective of the investigation was to develop a clinically valid three dimensional computer model of the orthodontic bracket-cement-tooth continuum, and determine the magnitude and distribution of stresses generated by three different load cases. A three-dimensional finite element model of the bracket-cement-tooth system was constructed consisting of 15,324 nodes and 2,971 finite elements. The stresses induced in the bracket-tooth interface by a masticatory load, a peel force and a twisting couple were recorded. The maximum principal stresses resulting from occlusal and 'twisting' forces are distributed toward the lute periphery. Peel forces, applied to the bracket tie wing, are concentrated beneath the bracket stem. Twisting forces result in the highest enamel stresses. The quality of orthodontic attachment can be explained by the magnitude and distribution of major principal stresses within the cement and impregnated bracket base. Shear and shear/peel forces are most likely to induce crack propagation within the adhesive layer. However, when a twisting action is used to remove orthodontic brackets, enamel failure is most likely. A clearer insight into the complexity of the bracket-cement-tooth system has been provided by numerical and finite element investigations. Further investigations, evaluating the influence of bracket base designs and orthodontic cement physical and geometric properties are indicated. Refereed Scientific Paper PMID- 10790444 TI - The reliability of the Index of Orthodontic Treatment Need over time. AB - The aim of this investigation was to establish whether the Index of Orthodontic Treatment Need is reliable over time between the ages of 11 and 19 years. It consisted of a longitudinal sample of 314 11- and 15-year-old and 142 19-year-old subjects who had not received orthodontic treatment or extractions. The changes in the aesthetic component (AC) and the dental health component (DHC) of the Index of Orthodontic Treatment Need (IOTN) were measured between the ages of 11 and 19 years. The results suggested that the dental health component of IOTN was reliable over time between the ages of 11-19 years despite temporal changes in the separate occlusal traits that comprise the index. The aesthetic component of IOTN tended to show an improvement over time. The Index of Orthodontic Treatment Need is a reliable index over time when taking into account occlusal changes that are occurring during the 11-19-year age range. The study provides some reassurance to clinicians that an IOTN grading at age 11 years is unlikely to change by the time the patient is 19 years. Refereed Scientific Paper PMID- 10790445 TI - The effect of food simulants on enamel-composite bond strength. AB - The effect of food simulants upon the enamel-composite bond strength of two orthodontic adhesive composites was studied. Thirty extracted premolars were used in each experimental group. Orthodontic brackets were bonded with either Transbond (3M Unitek) or Right-On (TP orthodontics) and were then exposed to either distilled water (control), 8 per cent ethanol (aqueous food), 50 per cent ethanol (alcoholic food), buffered lactic acid pH4 (acidic food), or corn oil (fatty food) for 12 weeks. Bond strength and Adhesive Remnant Index was recorded. It was found that 50 per cent ethanol had a significant effect upon the bond strength of both Transbond and Right-On. Right-On was affected to a greater extent. The clinical significance of this finding is that alcoholic mouth rinses, and alcohol-containing foods and drinks may be a causative factor in bond failure. However, the use of an alcoholic mouthrinse late in treatment may aid bracket removal. This potential use requires more research. Refereed Scientific Paper PMID- 10790447 TI - 'Journal of Orthodontics'--a change of outlook. PMID- 10790446 TI - Prevalence of signs and symptoms of temporomandibular disorders in young Nigerian adults. AB - The objective of this study was to determine the prevalence of signs and symptoms of TMD in young Nigerian adults and to establish a baseline for comparison with other studies. It consisted of a cross-sectional study at Obafemi Awolowo University, Ile-Ife, Nigeria in 1997. The subjects consisted of 308 medical and dental students (207 males and 101 females) randomly selected from a Nigerian University. Their ages ranged from 17 to 32 years with a mean age of 23 +/- 3.0 years. They were assessed according to the criteria of Helkimo (1974). Whilst 26.3 per cent of the subjects reported mild symptoms (Ai I) of TMD, only 2.9 per cent reported severe symptoms (Ai II). Similarly, 46 per cent showed mild dysfunction signs (Di I), whilst 16.5 and 0.3 per cent exhibited moderate (Di II) and severe (Di III) signs of TMD, respectively. No significant relationships were found between sex, anamnestic index, and the clinical dysfunction index scores. However, there were low but significant correlations between ananmestic index scores (Ai) and the recorded signs (Di), as well as the clinical dysfunction scores (CDS). There is some evidence to show that signs and symptoms of TMD occur amongst Nigerians, although restricted lateral and protrusive mandibular movements contributed significantly to clinical dysfunction scores. This report contrasts with what is found in western societies regarding the low prevalence of TMJ pain. Refereed Scientific Paper PMID- 10790448 TI - Reporting of clinical trials in the JO--the CONSORT Guidelines. PMID- 10790450 TI - A review of some orthodontic journals. PMID- 10790449 TI - Evidence-based orthodontics: where do I find the evidence? PMID- 10790451 TI - Series of reports on European orthodontics. The EURO-QUAL Biomed 2 Project. Original report series. PMID- 10790452 TI - A survey of undergraduate orthodontic education in 23 European countries. AB - This paper reports on a survey of teaching contents and time allocation within the undergraduate orthodontic curriculum in European countries in 1997, and on whether or not these countries set a formal undergraduate examination in orthodontics. A questionnaire and an explanatory letter were mailed to all members of the EURO-QUAL BIOMED II project. Answers were validated during a meeting of project participants and by fax when necessary. Completed questionnaires, which were subsequently validated, were returned by orthodontists from 23 countries. They indicated that orthodontics was taught in all undergraduate curriculums of the countries surveyed. The number of hours in the undergraduate curriculum devoted to orthodontics was reported as varying from 135 to 500 hours with a mean of 245 hours. The time reported as allocated to theory, clinical practice, laboratory work, diagnosis, and treatment planning varied widely. In general, clinical practice and theory were reported as being allocated most curriculum hours, whilst diagnosis, laboratory work, and treatment planing were reported as receiving relatively less time. Removable appliances were reported to be taught in 22 of the 23 countries, functional appliances in 21 countries and fixed appliances in 17 countries. An undergraduate examination in orthodontics was reported by 20 countries. It was concluded that orthodontics occupies a small proportion of the undergraduate curriculum in dentistry in most countries, the emphasis is on theory and clinical work, and that removable appliances, functional appliances, and certain aspects of fixed appliances are taught in the majority of countries that responded to the questionnaire PMID- 10790453 TI - A survey of postgraduate (specialist) orthodontic education in 23 European countries. AB - This paper reports on a survey of the duration, funding, and assessment of postgraduate specialist orthodontic training, the requirement for postgraduate training prior to entering specialist orthodontic training and registration of specialist orthodontists in Europe. A questionnaire and explanatory letter were mailed to all members of the EURO-QUAL BIOMED II project. Answers were validated during a meeting of project participants and by fax, when necessary. Completed questionnaires which were subsequently validated, were returned by orthodontists from 23 countries. The results indicated that a period of postgraduate training, prior to entering specialist orthodontic training was required in 12 of the responding countries. Specialist orthodontic training was reported as lasting 2 years in three countries, 3 years in 17, and for 4 years in three. Part-time training was reported as a possibility in four countries. In 21 of the 23 countries specialist training was reported to take place in full or part within universities, with some training taking place in government clinics in four countries. In five countries some or all training was reported to take place in specialist practices. Training was said to be funded solely or partially by governments in 15 of the 23 countries, to be solely self-funded in five countries, and partly or solely funded by universities in six countries. A final examination at the end of specialist training was reported to be held in 21 of the 23 countries. The nature of this examination varied widely and there was no such examination in two countries. Twelve of the 23 countries reported that they had a specialist register for orthodontics; 11 that they had no register. In none of the countries surveyed was there a requirement for those on a register to undergo periodic reassessment of competence once they are on the register. It was concluded that there was wide diversity in all aspects of specialist orthodontic training and registration within the countries surveyed. PMID- 10790454 TI - Orthodontics around the world: orthodontics in Brazil: excellence for a minority. AB - Brazil is the largest country of South America, with an area of 8.511.965 km(2) and 150 million people. It has 113 dental schools and several orthodontic postgraduate courses variously at Certificate, Master, and Doctoral levels. The current article gives an overview of the speciality in Brazil. The discussion puts the delivery of orthodontic care within the context of social conditions in Brazil. Included is a description of two full-time orthodontic courses located in the city of Rio de Janeiro. PMID- 10790455 TI - Alternating antipyretics: is this an alternative? AB - OBJECTIVES: To identify current fever management strategies and their basis, and to assess the frequency of alternating acetaminophen and ibuprofen. BACKGROUND: Practicing pediatricians confront the dilemma of elevated temperature and the anxiety this creates for parents. An informal survey of pediatricians revealed a variety of management strategies, including alternating acetaminophen and ibuprofen. There are no scientific data regarding alternating these 2 products. Design. During professional meetings, pediatricians voluntarily filled out a 15 item questionnaire. RESULTS: One hundred sixty-one completed surveys were reviewed. Respondents were mostly general pediatricians (67.7%), with >/=20 years in practice (55.9%). Most respondents chose a temperature of 101(o)F to start antipyretic treatment (61.9%). A small percentage used discomfort alone as the indication (13%). The antipyretic of choice was temperature-dependent in 50% of respondents; 57% used ibuprofen for temperature >/=102 degrees F. Fifty percent of respondents advised parents to alternate acetaminophen and ibuprofen. The method of alternation varied. The most common answers given for choosing a particular antipyretic regime were recommendations of the American Academy of Pediatrics (29%) and opinions of colleagues and mentors (25%). Physicians with <5 years of practice were significantly more likely to alternate antipyretics (69.7%). CONCLUSION: Acetaminophen and ibuprofen are commonly being used in an alternating manner for management of fever. There is presently no scientific evidence that this combination is safe or achieves faster antipyresis than either agent alone. There is evidence that the improper use of these agents may cause harm. Despite 29% of participants citing American Academy of Pediatrics recommendations as the basis for fever management, no such policy or recommendations exist. The observation that this practice is more common in younger practitioners may reflect their continued anxiety about fever (fever phobia). Until properly controlled studies have assessed the risk of combining these 2 products, practitioners should proceed with caution. PMID- 10790456 TI - Diminished inducible nitric oxide synthase expression in fulminant early-onset neonatal pneumonia. AB - OBJECTIVE: Fulminant early-onset neonatal pneumonia is associated with ascending intrauterine infection (IUI), prematurity, persistent pulmonary hypertension (PPHN), and septicemia. Nitric oxide (NO) as an inflammatory mediator is included in antimicrobial defense and has a role in pathogenesis of septic shock. The aim was to study the role of inflammatory NO in neonatal pneumonia. METHODS: Lungs from 36 autopsies were studied: 12 had fulminant early-onset neonatal pneumonia, 5 pneumonia of later onset, and 19 controls had similar gestational and postnatal age. In addition, airway specimens from 21 intubated newborns were analyzed: 7 with fulminant early-onset pneumonia, 7 apparently noninfected infants born prematurely attributable to IUI, and 7 premature infants of similar gestation. Specimens were analyzed for inducible NO synthase (NOS2) and nitrotyrosine, an indicator of NO toxicity. The degree of staining was analyzed. RESULTS: In fulminant pneumonia, alveolar macrophages (AM) showed significantly less NOS2 immunoactivity than the controls. In the airway specimens, the infants with fulminant pneumonia 0 to 2 days after birth had significantly lower intracellular NOS2 and nitrotyrosine and significantly lower interleukin-1beta and surfactant protein-A than apparently noninfected IUI infants. NOS2 and the other indices increased significantly during the recovery. CONCLUSIONS: For the first time, we report NOS2 expression by macrophages from human neonates. In fulminant early onset neonatal pneumonia, delayed production rather than excess of pulmonary inflammatory NO is associated with severe symptoms. PMID- 10790457 TI - Quality of care and use of the medical home in a state-funded capitated primary care plan for low-income children. AB - OBJECTIVE: To evaluate the quality of care and use of the medical home in a state funded capitated insurance plan for low-income children-the Colorado Child Health Plan (CCHP). DESIGN: A retrospective cohort study using medical record review at pediatric and family practice offices in 4 geographic areas of Colorado. At each practice, CCHP-enrolled children (6 months to 6.5 years) and 2 controls were selected, 1 with Medicaid (MK) and 1 with private insurance (PI), matched by date of birth to the CCHP-enrolled child (N = 596). CCHP-enrolled children with a diagnosis of asthma, aged 3 to 18 years, and asthmatic children with MK and PI, matched by age, were also selected from each practice (N = 139). RESULTS: Quality of preventive services were comparable in the 3 groups. CCHP-enrolled children made more health maintenance visits than MK-enrolled children (1.3 CCHP vs.9 MK vs 1.1 PI) and were more frequently screened for lead (8.1% CCHP vs 3.4% MK vs 1.2% PI) and anemia (5.0% CCHP vs 4.4% MK vs 2.4% PI) than children in either control group. Documented immunization rates were similar in the 3 groups, but a shift in location of immunization from public health clinics to the primary care site was seen in the CCHP group. CCHP-enrolled children made more office visits for acute care than did MK-enrolled children (4.1 CCHP vs 3.1 MK vs 3.4 PI), but a higher proportion of these visits took place at the medical home rather than the emergency department for the CCHP group (.04) as compared with the MK (.07) or PI (.06) groups. Asthmatic children in the CCHP group made more preventive office visits for maintenance therapy and more frequently used the primary care site rather than the emergency department for acute exacerbations than did children with PI (mean ratio of emergency department visits to total acute visits.04 CCHP vs.06 MK vs.19 PI). CONCLUSIONS: Despite capitated reimbursement for primary care services, CCHP provided children from low-income families with preventive, acute, and chronic care services of comparable quantity and quality to those received by children with MK or PI. The program was associated with a shift of immunization location to the primary care site and increased health maintenance care for new enrollees. CCHP-enrolled children used their medical home for the majority of acute health needs and were not high utilizers of emergency department or hospital services. PMID- 10790458 TI - Outcomes evaluation of a comprehensive intervention program for asthmatic children enrolled in medicaid. AB - OBJECTIVES: To evaluate health care and financial outcomes in a population of Medicaid-insured asthmatic children after a comprehensive asthma intervention program. DESIGN: Controlled clinical trial. SETTING: Pediatric allergy clinic in an urban, tertiary care children's hospital. SUBJECTS: Eighty children, 2 to 16 years old, with a history of frequent use of emergent health care services for asthma. Intervention. Children in the intervention group received asthma education and medical treatment in the setting of a tertiary care pediatric allergy clinic. An asthma outreach nurse maintained monthly contact with the families enrolled in the intervention group. OUTCOME MEASURES: Emergency department (ED) visits, hospitalizations, and health care charges per patient in the year after enrollment. RESULTS: Baseline demographics did not differ significantly between the 2 groups. In the year before the study, there were no significant differences between intervention and control children in ED visits (mean, 3.5 per patient), hospitalizations (mean,.6 per patient) or health care charges ($2969 per patient). During the study year, ED visits decreased to a mean of 1.7 per patient in the intervention group and 2.4 in controls, while hospitalizations decreased to a mean of.2 per patient in the intervention group and.5 in the controls. Average asthma health care charges decreased by $721/child/year in the intervention group and by $178/patient/year in the control group. CONCLUSIONS: A comprehensive asthma intervention program for Medicaid insured asthmatic children can significantly improve health outcomes while reducing health care costs.asthma education, health care outcomes, Medicaid, asthma outreach, utilization. PMID- 10790459 TI - Does candidemia predict threshold retinopathy of prematurity in extremely low birth weight (3 days of age, and 5 were discharged or transferred before ROP screening. A total of 449 infants were included in the study; 58 (13%) developed threshold ROP. Candidemia occurred in 58 (13%) infants before developing the worst stage of ROP. Candidemia occurred in 27 of 73 (37%) at 23 to 24 weeks' gestational age (GA), 25 of 197 (13%) at 25 to 26 weeks' GA, and 6 of 129 (5%) at 27 to 28 weeks' GA, 0 of 50 >28 weeks' GA. Similarly, threshold ROP occurred in 25 of 73 (34%) at 23 to 24 weeks' GA, 26 of 197 (13%) at 25 to 26 weeks' GA, and 6 of 129 (5%) at 27 to 28 weeks' GA, and 1 of 50 (2%) >28 weeks' GA. Threshold ROP developed in 19 of 58 (33%) infants with a history of candidemia and 39 of 391 (10%) without candidemia. Proportional hazards analysis indicated that GA in weeks (hazard ratio =.75; 95% confidence interval [CI]:. 61,.93) and non-black ethnicity (hazard ratio = 1.8; 95% CI: 1.05, 3. 08) were significantly associated with threshold ROP. After controlling for GA and other factors, candidemia did not remain significantly associated with threshold ROP (hazard ratio = 1.6; 95% CI:.89, 2.89). CONCLUSION: Candidemia may not be an independent risk factor for threshold ROP in extremely low birth weight infants. The magnitude of the previously reported association between candidemia and threshold ROP (more than fivefold) is unlikely and much of the clinically observed association appears to be mediated by gestational age. PMID- 10790460 TI - Systemic candidiasis in extremely low birth weight infants receiving topical petrolatum ointment for skin care: a case-control study. AB - BACKGROUND AND OBJECTIVE: An increase in the incidence of systemic candidiasis (SC) followed a change in skin care for extremely low birth weight (ELBW) infants in our neonatal intensive care unit (NICU). We sought to determine whether the use of topical petrolatum ointment (TPO) for skin care of ELBW infants was associated with risk for SC. STUDY DESIGN: Case-control study. SETTING: A 48-bed NICU in a private hospital in Houston, Texas. PATIENTS: Ten ELBW infants with and 30 without SC admitted to the NICU from December 1, 1997 through July 31, 1998. METHODS: ELBW infants with SC were identified using hospital microbiology and infectious disease consultation databases. A case was defined as an infant weighing 30 mL/kg (n = 29), a lower number of transfusions was received by infants in the early group, compared with late group, from the second week to the end of the treatment (early: 3.4 +/- 1.1 vs late: 5.4 +/- 3.7 transfusion/patient). No clinical adverse effects were observed. Thrombocytosis was detected during the treatment with rHuEPO in 31% of the infants. CONCLUSIONS: In the whole population, the early administration of rHuEPO induced a rise of reticulocyte counts, but not enough to reduce the transfusion requirement. The most severely ill infants (BW <800 g and phlebotomy losses >30 mL/kg) seemed to benefit from early use of rHuEPO, and this deserves additional study. PMID- 10790465 TI - Factors associated with age at operation for children with congenital heart disease. AB - OBJECTIVE: Previous studies have shown that children with congenital heart disease (CHD) who live in nonurban areas or who do not have private insurance are at risk for delayed referral to a pediatric cardiologist. However, the effect of these factors on the age at which cardiac surgery is performed has not been evaluated. This study is designed to evaluate the factors that influence the age at which definitive surgical repair is performed. METHODS: Data on hospital discharges for 1995 and 1996 in California were obtained from the Office of Statewide Health Planning and Development database. Children <18 years who underwent surgical repair for atrial septal defect (ASD), ventricular septal defect (VSD), tetralogy of Fallot (TOF), or atrioventricular canal (AVC) were included in the study. Age at surgery was evaluated using type of CHD, gender, race, type of insurance, surgical centers, urban or rural home location, and distance between home and surgical center as independent variables. RESULTS: In 1995-1996, 666 children underwent ASD closure (mean age: 5.1 years; median: 4.0 years), 582 VSD closure (mean age: 2.8; median: 1.1 years), 394 TOF repair (mean age: 1.7; median:.9 years), and 177 AVC repair (mean age: 1.1; median:.6 years). Comparing median and mean age at surgery, we found: AVC/=16) reported significantly poorer prevention practices for car seat use, covering electrical plugs, and having syrup of ipecac in the home. High depressive symptoms were also related to a lower likelihood of daily reading, but only for those mothers presently living with a male partner. Engagement in all prevention practices, except having syrup of ipecac in the home, were less likely if the mother reported high levels of depressive symptoms at both time points versus a single time point. CONCLUSION: Maternal depression may significantly impede parental prevention practices. As maternal depression is a treatable condition, screening and treating this disorder may contribute to improvement in childhood prevention practices and ultimately child health. PMID- 10790468 TI - Structural and functional brain integrity of fetal alcohol syndrome in nonretarded cases. AB - PURPOSE: To determine the structural and functional integrity of the brain in a sample of nonretarded individuals diagnosed with fetal alcohol syndrome. SUBJECTS: The sample consisted of nineteen individuals who met the diagnostic criteria for fetal alcohol syndrome. METHODS: Intellectual function was assessed using the Wechsler Adult Intelligence Scale-Revised. Structural integrity of the brain was assessed using magnetic resonance imaging whereas functional integrity was assessed using positron emission tomography and (18)F-fluorodeoxyglucose. RESULTS: The mean Full Scale IQ was 80. 2 (range: 66-92). Only 1 magnetic resonance imaging was found to be abnormal. This abnormality was found for the subject with the lowest IQ. Decreases in relative regional cerebral metabolic rates were found in 5 brain regions comprising the thalamus and basal ganglia. CONCLUSION: These results when coupled with previous findings suggest a continuum of neuropathology in fetal alcohol syndrome. For cases with relatively mild intellectual deficits, the cause of the deficit is at a micro-level rather than a macro-level. PMID- 10790469 TI - Electrocardiographic findings in adolescents with eating disorders. AB - OBJECTIVES: We sought to determine the prevalence and clinical correlates of electrocardiographic findings in adolescents with eating disorders. STUDY DESIGN: We undertook matched case-control study of electrocardiographic findings at initial assessment in 62 adolescents with anorexia nervosa, 9 with bulimia nervosa, and 26 with eating disorder not otherwise specified presenting from March 1995 to September 1996. RESULTS: Mean (+/- 1 standard deviation) age was 15.0 +/- 1.4 years (95% were female). Patients with anorexia nervosa had significantly lower heart rates (mean case-control difference: -20 +/- 17 beats per minute), lower R in V(6) (-2.6 +/- 5.5 mm), longer QRS interval (+.004 +/ .010 seconds), shorter mean QTc (-.0136 +/-.033 seconds) and lesser QTc dispersion (-.010 +/-. 031 seconds). The bulimia nervosa group had slightly longer mean QTc (.019 +/-.020 seconds), with no significant case-control differences in the eating disorder not otherwise specified group. CONCLUSION: Electrocardiographic findings are abnormal in adolescents with anorexia nervosa but not in adolescents with bulimia nervosa or eating disorder not otherwise specified. PMID- 10790470 TI - Echocardiographic predictors of outcome in newborns with congenital diaphragmatic hernia. AB - OBJECTIVE: Despite new treatments, congenital diaphragmatic hernia (CDH) still has high mortality. The aim of this study was to identify echocardiographic predictors of outcome in newborns with an isolated CDH. METHODS: We reviewed medical charts and echocardiograms of 40 newborns who presented with CDH in the first 24 hours of life, from 1992 to 1996. We compared the cardiac valves and great arteries diameters, left-ventricular volume and mass, Apgar scores, and modified McGoon index (the combined diameter of hilar pulmonary arteries, indexed to the descending aorta) of survivors and nonsurvivors. We performed Student's t test and multiple logistic regression analysis between the 2 groups. RESULTS: Fourteen patients died 1 to 33 days after birth (median: 3 days), including 8 from progressive hypoxemia without operation; 26 have survived up to 5 years (median: 2 years) after successful operations. Nonsurvivors had significantly smaller diameters of right and left hilar pulmonary arteries, more frequent right sided CDH, and lower mean Apgar scores at 1 and 5 minutes. The most significant prognostic factor was the modified McGoon index. A modified McGoon index 0.05) between sites, although activities of benzyloxy-O-dealkylase and ethoxyresorufin-O-dealkylase were somewhat elevated in 3 of the 15 embryos collected from Pea Patch Island. Concentrations of 21 organochlorine pesticides and metabolites were relatively low at both sites, with p,p'-DDE values well below the threshold associated with eggshell thinning. Although total PCB concentration was modestly elevated (p < 0.05) in Pea Patch Island heron embryos, levels of arylhydrocarbon receptor-active PCB congeners, polychlorinated dibenzodioxins and dibenzofurans, and toxic equivalents were low and did not differ between sites. Concentrations of Cd and Mn in pipping embryos from Pea Patch Island were slightly greater (p < 0.05) than values observed in Middle Island embryos, but levels of these and the other metals and metalloids (e.g., Hg and Se) were below values associated with toxicity. In conclusion, it seems unlikely that chlorinated hydrocarbon and metal contaminant exposure constitutes a direct threat to the reproductive success of black-crowned night herons at Pea Patch Island. However, low-level exposure to these contaminants may constitute one of many stressors that in combination could adversely affect the stability of the wading bird population at this large heronry. PMID- 10790502 TI - A survey of size-specific mercury concentrations in game fish from Maryland fresh and estuarine waters. AB - A survey of size-specific mercury (Hg) concentrations in game fish from a subset of Maryland fresh and estuarine waters was conducted, in which Hg concentrations in 112 fish from seven freshwater impoundments and three tidal and four estuarine locations in Chesapeake Bay and its tributaries were measured. Striped bass (Morone saxatilis) was the most intensively examined species. Of the fish examined, the largest freshwater sportfish contained the highest Hg concentrations. Striped bass and largemouth bass (Micropterus salmoides) from Chesapeake Bay and its tributaries contained less Hg at the same size than the same species in fresh waters. Large striped bass, chain pickerel (Esox niger), and walleye (Stizostedion vitreum) from Deep Creek Lake and Liberty Reservoir exceeded the FDA action level of 1 mg Hg/kg. Striped bass, largemouth bass, and white crappie (Pomoxis annularis) in other impoundments equaled or exceeded a common advisory level of 0.5 mg Hg/kg. Large differences in size-normalized Hg concentrations among lakes and particularly between fresh and salt waters highlight the large differences in MeHg production and bioaccumulation among ecosystems. This work indicates that a more comprehensive study of Hg in Maryland fish is warranted to protect human and wildlife health. PMID- 10790501 TI - The effect of bioaccumulated selenium on mortalities and deformities in the eggs, larvae, and fry of a wild population of cutthroat trout (Oncorhynchus clarki lewisi). AB - To determine if elevated concentrations of waterborne selenium (Se), caused by coal mining, in the Elk River in southeastern British Columbia, may be causing reproductive or teratogenic effects in wild cutthroat trout (Oncorhynchus clarki lewisi), fertilized eggs from exposed and reference fish were raised in the laboratory. Eggs from each female were reared separately and the percent mortalities and deformities were related to the selenium content of the eggs. Selenium concentrations in females from the exposed site were highest in the liver (36.6 +/- 22.5 microg/g dry weight, range: 18.3 to 114), followed by the eggs (21.0 +/- 18.3 microg/g, range: 8.7 to 81.3) and the muscle (12.5 +/- 7.7 microg/g, range: 6.7 to 41). Despite these elevated egg Se concentrations, there was no significant effect on fertilization; time to hatch; percent hatch; or egg, larvae, and fry deformities or mortalities. Reproductive failure and embryonic terata have been reported at much lower egg Se concentrations in other fish species. The lack of any toxic response in this study may be due to an evolved tolerance to higher tissue Se concentrations in a population of fish living in a seleniferous river system. PMID- 10790503 TI - Soil dissipation of diuron, chlorotoluron, simazine, propyzamide, and diflufenican herbicides after repeated applications in fruit tree orchards. AB - In a pear tree orchard planted on loam soil, each plot was treated in April 1998 with either one of the ureas diuron or chlorotoluron, or triazine simazine herbicides applied at 3, 4, and 2 kg AI ha(-1), respectively. Some plots had not been previously treated with one of these herbicides. Other plots had been treated annually during the past 12 years with the same herbicide. One herbicide, and always the same, was thus applied to each plot. In the plots treated for the first time with either diuron, chlorotoluron, or simazine, the soil half-lives of these herbicides in the 0-10 cm surface soil layer were 81, 64, and 59 days, respectively. In the plots treated with the same herbicide for 12 years, the corresponding soil half-lives were 37, 11, and 46 days. Diuron thus produced a moderately enhanced biodegradation, chlorotoluron a high one, and simazine a low but significant one. In another pear tree orchard planted on sandy loam soil, each plot was treated in April 1998 with one of the amide propyzamide (1.25 or 1.0 AI kg ha(-1)) or diflufenican (250 g AI ha(-1)) herbicides. In the plots not previously treated with propyzamide, the propyzamide soil half-life was the same for both doses, i.e., about 30 days. In the plots treated annually for 3 or 14 years with propyzamide, the soil half-life was 12 and 10 days, respectively. In the plots treated for the first time with diflufenican and in those treated annually with diflufenican for 3 years, the diflufenican soil half-life was the same, i.e., 65 days. Propyzamide thus already showed a highly accelerated biodegradation after 3 years of repeated annual applications. Diflufenican, however, did not show enhanced biodegradation after 3 years of repeated annual applications. PMID- 10790504 TI - Chemistry of rain water in Riyadh, Saudi Arabia. AB - Twenty-three rain water samples were collected from 15 sampling locations in Riyadh during four rain events in 1994 and analyzed for pH, conductivity, TDS, alkalinity, Cl, NO(3), F, SO(4), Na, Mg, Ca, and 12 trace metals. The pH ranged between 6.83 and 8.21. The major cation and anion were Ca and SO(4), contributing with 30.5% and 20. 8%, respectively, to TDS. Moderate variation in major ions concentration was found to occur between different sites. The concentrations of different metals varied considerably from one sampling location to another. The dominant trace metal ions in the rain water samples were Zn, Pb, and Al. Samples collected from a site near a cement factory in Riyadh showed maximum concentrations of most of the major ions and metals. The concentrations of various ions in the first rain event were comparatively higher than the concentrations of these ions in the later rain events. PMID- 10790505 TI - The effect of pH on the kinetics of intra- and extracellular uptake of Al in Fontinalis antipyretica: changes in the cellular contents of K, Mg, and Ca. AB - The effect of acidity on the accumulation of Al in the aquatic bryophyte Fontinalis antipyretica was studied. The main interest of this study was in characterizing the relationship between the two parameters, as Al is typically mobilized in acidic environments, where it becomes extremely toxic. With this aim, a series of laboratory experiments were carried out where samples of F. antipyretica were held in tanks of water with different concentrations of Al (0.1 3 mg L(-1)) and levels of acidity (pH 3-5. 8). The incubation time varied between 1 and 24 h. In general, the greatest accumulations of both intra- and extracellular Al were found at a pH of 4.4. With the results obtained, the kinetics of both intra- and extracellular Al uptake was characterized, and models constructed for different combinations of Al and acidity. The study was completed by examining the relationship between the pH of the holding water and bioaccumulation of Al and the variation in concentration of cations essential to plant life, which are also indicators of changes in cellular physiology (K, Mg, Ca). In general, losses of these elements were found at intra- and extracellular locations, especially at high pH. PMID- 10790506 TI - Increased abundance of terrestrial isopod populations in terrestrial ecosystems contaminated with petrochemical wastes. AB - Arthropods are integral components of the food chain, and because many reside in close association to the soil and its contaminants, their communities may be sensitive indicators of ecotoxicity. We examined the influence of petrochemical contaminants on the abundance of several taxonomic groups of macroarthropods, with an emphasis on isopods, residing on two reference sites and three petrochemical-contaminated sites at an abandoned oil refinery site in southwestern Oklahoma. Relative densities of surface-dwelling macroarthropod assemblages were significantly greater on contaminated sites than reference sites. Differences in terrestrial isopod populations were particularly remarkable and consistent across study sites, with densities approaching 180-fold greater on contaminated than reference sites. Similarity comparisons indicated that abundances and composition of macroarthropod assemblages sampled on the contaminated sites were more similar to one another than to the communities residing on reference sites. Although preliminary, these initial results support the hypothesis that macroarthropod assemblages, in particular isopods, are sensitive to soil contaminants and could be a valuable approach to evaluating the effects of petrochemical contamination of soils on terrestrial ecosystems. PMID- 10790507 TI - Long-term effects of pesticide exposure at various life stages of the southern leopard frog (Rana sphenocephala). AB - Amphibian larvae are commonly exposed to low levels of pesticides during their development. Chronic studies generally examine the effects of long-term exposure, but they often disregard the importance of the individual life stage at which tadpoles are exposed. I determined the point during development at which carbaryl effects are manifested by exposing southern leopard frog tadpoles (Rana sphenocephala) to the pesticide carbaryl at five different times during development. Metamorphs exposed throughout the tadpole stage and throughout development (egg, embryo, tadpole) experienced significant mortality at all chemical levels. Although the length of the larval period was the same for all experimental groups, metamorphs exposed during the egg stage were smaller than their corresponding controls, independent of whether they were exposed at any other stage. Nearly 18% of individuals exposed to carbaryl during development exhibited some type of developmental deformity (including both visceral and limb malformities), compared to a single deformed (< 1%) control tadpole, demonstrating that a chemical hypothesis for amphibian deformities remains viable. Because exposure to nonpersistent chemicals may last for only a short period of time, it is important to examine the long-term effects that short-term exposure has on larval amphibians and the existence of any sensitive life stage. Any delay in metamorphosis or decrease in size at metamorphosis can impact demographic processes of the population, potentially leading to declines or local extinction. PMID- 10790508 TI - Field evaluation of lead effects on Canada geese and mallards in the Coeur d'Alene River Basin, Idaho. AB - Hatch year (HY) mallards (Anas platyrhynchos) in the Coeur d'Alene (CDA) River Basin had higher concentrations of lead in their blood than HY Western Canada geese (Branta canadensis moffitti) (geometric means 0.98 versus 0.28 microg/g, wet weight). The pattern for adults of both species was similar, although geometric means (1.77 versus 0. 41 microg/g) were higher than in HY birds. HY mallards captured in the CDA River Basin in 1987 contained significantly lower lead concentrations in their blood than in 1994-95 (0.36 versus 0.98 microg/g); however, some very young mallards were sampled in 1987, and concentrations in adults were not significantly different in 1987, 1994, or 1995 (1.52, 2.07, 1.55 microg/g, respectively). Both species in the CDA River Basin in 1994-95 showed significantly reduced red blood cell delta-aminolevulinic acid dehydratase (ALAD) activity compared to the reference areas: Canada geese (HY -65.4 to -86.0%, adults -82.3%), and mallards (HY -90.7 to -95.5%, adults -94. 1%). Canada goose goslings were divided into size classes, and the two smaller classes from the CDA River Basin had significantly elevated free erythrocyte protoporphyrin (protoporphyrin) levels compared to the reference area (15.2x and 6.9x). HY and adult mallards both had significantly elevated protoporphyrin (5.9x and 7. 5x). Recognizing that interspecific differences exist in response and sensitivity to lead, it appears (at least for hemoglobin and hematocrit) that Canada geese were more sensitive to lead than mallards, i.e., adverse hematologic effects occur at lower blood lead concentrations. Only Canada geese from the CDA River Basin, in spite of lower blood lead concentrations, had significantly reduced mean hemoglobin and hematocrit values. No euthanized Canada geese (all HYs) from CDA River Basin were classified as clinically lead poisoned, but 38 Canada geese found dead in the CDA River Basin during a concurrent study succumbed to lead poisoning between 1992 and 1997. Only 6 (15.8%) of these 38 contained ingested lead shot, which contrasts greatly with the 75-94% incidence of ingested lead shot when mortality was due to lead shot ingestion. Lead from other contaminated sources (i.e., sediments and vegetation) in the CDA River Basin was strongly implicated in most Canada goose deaths. Based on the 31 live mallards and Canada geese collected in the CDA River Basin, which were representative of the live populations blood sampled only, the prevalence of subclinical and clinical lead poisoning (as determined by liver lead concentrations, excluding birds with ingested lead shot) was higher in mallards: subclinical (4 of 8, 50% HYs and 6 of 11, 55% adults); clinical (0% HYs and 4 of 11, 36% adults), with less data available for Canada geese (only 1 of 9, 11% HYs marginally subclinical). The clinically lead-poisoned mallards had extremely high concentrations of lead in blood (2.69-8. 82 microg/g) and liver (6.39-17.89 microg/g). Eight mallards found dead in the CDA River Basin during a concurrent study were diagnosed as lead poisoned, and only one (12.5%) contained ingested lead shot, which again strongly implicates other lead sources. The finding of dead lead poisoned Canada geese together with the high percentage of live mallards classified as subclinically or clinically lead poisoned, in combination with the low incidence of ingested lead shot causes us concern for both of these species, which live in association with lead-contaminated sediment in the CDA River Basin. PMID- 10790510 TI - Variability of mercury and selenium levels in clutches of Audouin's gulls (Larus audouinii) breeding at the Chafarinas Islands, Southwest Mediterranean. AB - We determined mercury and selenium in 43 eggs (eggshell, albumen, and yolk) which belong to different clutch sizes of Audouin's gull from the Chafarinas Islands. The results were compared with those obtained previously with the same species at the Ebro Delta. Both, the intra- and the interclutch sources of variability have been examined. There is an effect of the female on mercury and selenium concentrations in a clutch, which supports the use of eggs as monitoring tools. The distribution pattern of mercury among albumen, yolk and eggshell, the dynamics of this element during the laying process, as well as data concerning egg formation strategies suggest that the mercury in the albumen corresponds mainly to the mercury acquired by the female while feeding in the breeding area. The mercury and selenium levels of the eggs from the Chafarinas Islands were lower than those of the Ebro Delta, which can be due to differences in both the marine contamination and the diet in the two colonies. PMID- 10790509 TI - Utility of Caenorhabditis elegans for assessing heavy metal contamination in artificial soil. AB - There is an increasing need for the development of soil bioassay protocols. Currently the only internationally standardized soil test organism is the lumbricid earthworm Eisenia fetida. Many alternate soil test organisms have been proposed. This work compares Caenorhabditis elegans to several other test organisms, including E. fetida, for heavy metals in soil. In this evaluation, such factors as ease of testing and culturing, duration of testing, soil volume needed, and the sensitivity of the organism were considered. Results show that C. elegans is more sensitive than most other organisms evaluated and is similar in response to E. fetida. The second issue compares C. elegans LC(50) values to heavy metals criteria specified in the US EPA regulations for land application of sewage sludge. Currently, the regulations are set on total metals in the soil and do not consider bioavailability of the metals. Regulations do not consider soil physiochemical properties, such as organic matter content, clay content, and cation exchange capacity, which have been shown to affect the availability of metals to soil organisms. While the C. elegans LC(50) values are above standard values in artificial soil, work in our lab indicates that the LC(50)s are below regulation values for other soil types. Due to the ease of culturing and testing, good sensitivity, along with the wealth of biological information and ecological relevance, C. elegans is a good organism for use in soil bioassays. PMID- 10790511 TI - Effects of mercury on the isolated perfused rat tail vascular bed are endothelium dependent. AB - The effects of mercury on vascular smooth muscle results in vasoconstriction, but the mechanism of this action is not elucidated yet. To investigate this issue we examined the effects of HgCl(2) in the isolated rat tail vascular bed. The tail artery was dissected, cannulated, and perfused at a constant flow (2.5 ml/min) with Krebs solution plus EDTA 0.03 mM at 36 degrees C. After equilibration for 30 min the effects of increasing concentrations of HgCl(2) (0.5, 1, 2, 5, and 10 microM) on the perfusion pressure were investigated. Concentrations of HgCl(2), 2 microM and above, significantly increased perfusion pressure. Blockade of alpha receptors (prazosin 84 ng/ml) did not alter the responses to HgCl(2), suggesting that the metal does not induce the release of neurotransmitters from sympathetic nerve terminals. To investigate the possible role of endothelium on the vasoconstriction produced by HgCl(2), preparations were precontracted with 10(-7) M phenylepherine or perfused with 5 microM HgCl(2) for 20 min. Acetylcholine vasodilated preparations precontracted with phenylepherine demonstrating the integrity of the endothelial nitric oxide-releasing mechanism. In contrast, after perfusion with 5 microM HgCl(2), the vasodilation produced by acetylcholine was abolished. In the presence of either phenylephrine or HgCl(2) the effects of sodium nitroprusside remained unchanged. Pretreatment with 30 microM indomethacin fully prevented the HgCl(2)-induced vasoconstriction. However, the endothelium dependent vasodilation in response to acetylcholine was significantly reduced after indomethacin plus HgCl(2) treatment, meanwhile the vasodilation produced by nitroprusside remained unchanged. Pretreatment with L-arginine (1 mM) did not prevent the vasoconstriction induced by HgCl(2), nor did it restore the ability of acetylcholine to produce vasodilation, and it did not alter the response to sodium nitroprusside. The possibility of HgCl(2)'s actions mediated by the formation of free radicals was also investigated. The administration of 10 mM histidine significantly reduced the vasoconstrictor response if used before HgCl(2) treatment without improving the reduced vasodilation produced by acetylcholine. These results are consistent with the hypothesis that the vasoconstriction produced by HgCl(2) may be mediated by the formation of superoxide anions, stimulating the production of a COX-derived vasoconstrictor agent and by reducing the endothelial vasodilator activity. PMID- 10790512 TI - Plant Species and Plant Incubation Conditions Influence Variability in Epiphytic Bacterial Population Size. AB - The influences of plant species and plant incubation conditions on the variability in bacterial population sizes among leaves were investigated in field and growth chamber studies. Pseudomonas syringae strains TLP2 and Cit7 were inoculated onto plants and population sizes were measured at intervals after inoculation. Total bacterial population sizes were also assessed in field studies. Levels of leaf-to-leaf variability in both P. syringae population size and bacterial community size differed significantly among plant species. For all plant species, variability among leaves in population sizes of inoculated bacteria was consistently greater than the leaf-to-leaf variability in numbers of total bacteria. Considering levels of variability in population size immediately prior to and following incubation under either wet or dry physical conditions, leaf-to-leaf variability in the population sizes of inoculated P. syringae strains increased significantly following incubation under dry, but not under wet, conditions. Measurements of leaf-to-leaf variability immediately prior to and following incubation were positively correlated regardless of whether the incubation was under wet or dry conditions, though the correlation was greater following dry incubation. These data provide insight into the biological and physical factors that may be important in generating variability in bacterial population sizes among leaves, and they have important implications for the design of appropriate strategies for sampling leaf surface microbial populations. 10 y) nutrient addition plots. Nitrogen fixation activity was assayed in each short-term treatment and control plot using an acetylene reduction technique. The control and nutrient addition DGGE profiles were very similar throughout the short-term experiments. One DGGE band that was prominent in the control plots was not found in the long-term nutrient addition plots. Diazotrophy may provide a competitive advantage for some species in this system, as indicated by results from the long-term nutrient amended plots. However, the rhizosphere environment in situ appears to limit the immediate impacts of increased nutrient availability on the diazotroph assemblage composition. Results from the short term nutrient amended plots showed no measurable effect on the diazotroph assemblage. These results indicate substantial short-term stability of the diazotroph assemblage composition, but the potential for change in the face of long-term changes in nutrient availability. 95 kg, 85+/-22 min. Previous abdominal surgery did not affect operative time. Nine patients (16%) required conversion from 2- to 5-mm ports because of adhesions, wall thickening, or need for better retraction. Time in these patients was 95+/ 26 min vs. 68+/-21 min in other patients (p<0.01). No patient was converted to an open procedure. Three patients (5%) had a positive cholangiogram and common bile duct exploration that required placement of an extra 5-mm trocar. Five patients (9%) required insertion of an additional 2-mm port. All patients received patient controlled analgesia (PCA). Morphine use was 0.21+/-0.19 mg/kg (range, 0-0.8). Hospital stay was 1.31 days (range, 0.5-4). Subjective satisfaction was excellent because of smaller incisions. No additional morbidity was seen with MLC. CONCLUSION: MLC is a feasible and safe approach that provides similar times to SLC with better cosmesis, a less painful recovery, and possibly an earlier return to normal activity. PMID- 10790551 TI - Pain after microlaparoscopic cholecystectomy. A randomized double-blind controlled study. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is traditionally performed with two 10-mm and two 5-mm trocars. The effect of smaller port incisions on pain has not been established in controlled studies. METHODS: In a double-blind controlled study, patients were randomized to LC or cholecystectomy with three 2-mm trocars and one 10-mm trocar (micro-LC). All patients received a multimodal analgesic regimen, including incisional local anesthetics at the beginning of surgery, NSAID, and paracetamol. Pain was registered preoperatively, for the first 3 h postoperatively, and daily for the 1st week. RESULTS: The study was discontinued after inclusion of 26 patients because five of the 13 patients (38%) randomized to micro-LC were converted to LC. In the remaining 21 patients, overall pain and incisional pain intensity during the first 3 h postoperatively increased in the LC group (n = 13) compared with preoperative pain levels (p<0.01), whereas pain did not increase in the micro-LC group (n = 8). CONCLUSIONS: Micro-LC in combination with a prophylactic multimodal analgesic regimen reduced postoperative pain for the first 3 h postoperatively. However, the micro-LC led to an unacceptable rate of conversion to LC (38%). The micro-LC instruments therefore need further technical development before this surgical technique can be used on a routine basis for laparoscopic cholecystectomy. PMID- 10790552 TI - Prospective randomized blinded trial of pulmonary function, pain, and cosmetic results after laparoscopic vs. microlaparoscopic cholecystectomy. AB - BACKGROUND: The size of laparoscopic instruments has been reduced for use in abdominal video endoscopic surgery. However, it has yet to be proven that microlaparoscopic surgery will actually result in clinically relevant benefits for patients. METHODS: Fifty patients were randomized in a blinded fashion to receive either elective laparoscopic (MINI), (n = 25) or microlaparoscopic (MICRO) (n = 25) cholecystectomy. Pulmonary function (FVC, FEV (1)), analgesic consumption during patient-controlled analgesia (PCA), pain perception by visual analogue score (VAS), and the cosmetic result (by the patient's self-assessment) were evaluated postoperatively as clinically relevant end points. RESULTS: Age, sex, body mass index (BMI), preoperative pulmonary function, pain perception, and operative time were similar for the two groups. At 8:00 PM on the day of surgery, FVC (MINI: 1.96 L [range, 1.48-2.48]; MICRO: 2.13 L) [(range, 1.61.-2.50)] and FEV (1) (MINI: 1.17 L/sec) [range, 0.87-1. 48]; MICRO: 1.34 L/sec [range, 1.05. 2.14] were also similar (each p = 0.5). From surgery to the 3rd postoperative day, cumulative PCA morphine doses were comparable (MINI: 0.15 mg/kg bw [range, 0.09-0. 23]; MICRO: 0.21 mg/kg bw [range, 0.10-0.42]; p = 0.4), but overall VAS scores for pain while coughing were higher in the laparoscopic group (406 [range, 358-514]) than in the microlaparoscopic group (365 [range, 215-427]; p = 0.02). The cosmetic result was judged to be slightly superior by the microlaparoscopic patients (10 [range, 9-10]), as compared to those in the laparoscopic (9 [range, 8-10]) group (p = 0.04). CONCLUSION: Because microlaparoscopic cholecystectomy has some minor advantages over laparoscopic surgery, it should be considered for use in selected patients. PMID- 10790553 TI - Minimally invasive surgery for posterior gastric stromal tumors. AB - BACKGROUND: Because involvement is extremely rare, surgery for gastric stromal tumors consists of local excision with clear resection margins. The aim of this study was to report the results of a consecutive series of nine patients with posterior gastric stromal tumors that were excised using a minimally invasive method. METHODS: Patients received a general anesthetic before placement of three laparoscopic ports- a 10-mm (umbilical) port for the telescope and two working ports, a 12-mm port (left upper quadrant) and a 10-mm port (right upper quadrant). Grasping forceps were placed through an anteriorly placed gastrotomy to deliver the tumor through the gastrotomy into the abdominal cavity, thus allowing an endoscopic linear cutter to excise the tumor with a cuff of normal gastric tissue. RESULTS: Nine consecutive patients with a median age of 73 years (range, 47-83) were treated. In seven patients, laparoscopic removal of the tumor was achieved. Two patients required conversion to an open operation because the tumor could not be delivered into the abdominal cavity. The median length of postoperative stay for the seven patients in whom the procedure was completed laparoscopically was 3 days (range, 2-6). CONCLUSIONS: Posterior gastric stromal tumors can be removed safely using this minimally invasive method. Delivery of the tumor through the gastrotomy is essential for success. PMID- 10790554 TI - More than two structures in Calot's triangle. A postmortem study. AB - BACKGROUND: Large laparoscopic cholecystectomy series often fail to report the rate at which a third structure is encountered in Calot's triangle. METHODS: During a 6-month period, the liver and hepatoduodenal ligament of 90 consecutive human cadavers underwent corrosion casting (n = 50), postmortem arteriography (n = 20), and postmortem cholangiography (n = 20). RESULTS: Third structures within Calot's triangle were arteries (0.6-5.7 mm diameter) in 36.2% (early division of the right hepatic artery, 8.6%; caterpillar hump right hepatic artery, 12.9%; liver branch of the cystic artery, 10%; double cystic arteries, 5.7%), bile ducts (0.3-1.6 mm diameter) in 5. 7% (small-caliber sectoral ducts, 1.4%; right posterior hepatic ducts, 4.3%), and veins (0.9-1.6 mm diameter) merging with the portal vein in 4% of the specimens. CONCLUSION: Knowledge of the aforementioned anatomy is critical to surgeons facing more than two structures within Calot's triangle during laparoscopic cholecystectomy. PMID- 10790555 TI - Laparoscopic cholecystectomy in acute cholecystitis. A prospective comparative study in patients with acute vs. chronic cholecystitis. AB - BACKGROUND: The aim of this prospective study was to compare the outcome of laparoscopic cholecystectomy (LC) in patients with acute cholecystitis versus those with chronic cholecystitis and to determine the optimal timing for LC in patients with acute cholecystitis. METHODS: From January 1991 to July 1998, 796 patients (542 women and 254 men) underwent LC. In 132 patients (67 women and 65 men), acute cholecystitis was confirmed via histopathological examination. These patients were divided into two groups. Group 1 (n = 85) had an LC prior to 3 days after the onset of the symptoms of acute cholecystitis, and group 2 (n = 47) had an LC after 3 days. RESULTS: There were no mortalities. The conversion rates were 38.6% in acute cholecystitis and 9.6% in chronic cholecystitis (p<10(-8)). Length of surgery (150.3 min vs. 107.8 min; p<10(-9)), postoperative morbidity (15% vs. 6.6%; p = 0.001), and postoperative length of stay (7.9 days vs. 5 days; p< 10( 9)) were significantly different between LC for acute cholecystitis and elective LC. For acute cholecystitis, we found a statistical difference between the successful group and the conversion group in terms of length of surgery and postoperative stay. The conversion rates in patients operated on before and after 3 days following the onset of symptoms were 27% and 59.5%, respectively (p = 0.0002). There was no statistical difference between early and delayed surgery in terms of operative time and postoperative complications. However, total hospital stay was significantly shorter for group 1. CONCLUSIONS: LC for acute cholecystitis is a safe procedure with a shorter postoperative stay, lower morbidity, and less mortality than open surgery. LC should be carried out as soon as the diagnosis of acute cholecystitis is established and preferably before 3 days following the onset of symptoms. Early laparoscopic cholecystectomy can reduce both the conversion rate and the total hospital stay as medical and economic benefits. PMID- 10790556 TI - Effects of pneumoperitoneum on cardiac autonomic nervous activity evaluated by heart rate variability analysis during sevoflurane, isoflurane, or propofol anesthesia. AB - BACKGROUND: The effects of pneumoperitoneum on the activity of the cardiac autonomic nervous system have not been completely understood. METHODS: In this study, 45 unpremedicated adult patients who underwent laparoscopic cholecystectomy were anesthetized with either 3.5% sevoflurane, 2% isoflurane, or 8 mg/kg/h propofol (15 patients in each group). The status of cardiac autonomic nervous activity was evaluated by heart rate variability analysis three times: once when the patient was awake, once after induction of general anesthesia, and once after insufflation for pneumoperitoneum. Intra-abdominal pressure was maintained automatically at 10 mm Hg by a carbon dioxide (CO(2)) insufflator. For each measurement, electrocardiogram was recorded for 256 s and played back offline to detect R-R intervals. Power spectral analysis of heart rate variability was applied, and the low-frequency (LF, 0.04-0.15 Hz) and high frequency (HF, 0.15-0. 40 Hz) bands of the spectral density of the heart rate variability were obtained from a power spectra of R-R intervals using the fast Fourier transform algorithm. The HF/LF ratio also was analyzed. RESULTS: Measurements of heart rate variability in the three groups showed similar change. Although the power of HF, which represents parasympathetic nervous activity, did not change, the power of LF, which represents both sympathetic and parasympathetic nervous activity, decreased during the anesthetized stage and increased during the insufflated stage. The HF/LF ratio, which represents the balance of parasympathetic and sympathetic activity, increased after induction of general anesthesia, and decreased after insufflation. CONCLUSIONS: Our results suggest that pneumoperitoneum increases sympathetic cardiac activity. The choice of general anesthetic did not seem to have a major influence on the change in the cardiac autonomic nervous system after induction of pneumoperitoneum for laparoscopic cholecystectomy. PMID- 10790557 TI - Effect of pressure and gas type on intraabdominal, subcutaneous, and blood pH in laparoscopy. AB - BACKGROUND: According to the literature, the number of port-site metastases in laparoscopic surgery varies considerably depending on the type of gas used for the pneumoperitoneum. In order to investigate this observation we studied the changes in blood, subcutaneous, and intra-abdominal pH during laparoscopy with helium, CO(2) and room air in a rat model. In addition, we looked at the influence of intra-abdominal pressure and duration of pneumoperitoneum on the pH during the laparoscopy. METHODS: pH was measured by tonometry, intra-abdominally and subcutaneously. A pH electrode was additionally placed into the subcutaneous tissue and the results compared to those measured by tonometry. Blood samples were taken from a catheter in the carotid artery. The intra-abdominal pressure was 0, 3, 6, 9 mm Hg for 30 min in each case. We investigated the effect of pneumoperitoneum with CO(2), helium and air in randomized groups of 5 rats. In an additional series the pressure was held constant at 3 mm Hg and the pH was measured every 30 min. RESULTS: Due to the different absorption capacity of the peritoneum, laparoscopy with CO(2) decreases the subcutaneous pH from 7.35 to 6.81. Blood pH is reduced from 7.37 to 7.17 and the intra-abdominal pH from 7.35 to 6.24. Other, less absorbable gases induce smaller changes of blood and subcutaneous pH (only 10% of CO(2)). In a variance analysis the p value is less than 0.001. The influence of duration of laparoscopy (30 min vs. 90 min) on the subcutaneous pH is less compared to the influence of intra-abdominal pressure (0, 3, 6, 9 mm Hg). CONCLUSIONS: Depending on the type of gas (CO(2), air, helium) used for laparoscopy blood, subcutaneous and intra-abdominal pH are influenced differently. Because lower pH is known to impair local defense mechanisms, these results may be one explanation for the higher incidence of port-site metastasis in laparoscopy with CO(2) than with other gases, as reported in the literature. PMID- 10790560 TI - The Tubingen balloon. A new method for adjusting the tension of the fundic wrap during laparoscopic Nissen fundoplication. AB - An adequate fundic wrap is fundamental to the success of conventional and laparoscopic Nissen fundoplications. Nevertheless, up to now there has been no standardized method for the surgeon to determine intraoperatively the width and tension of the fundic wrap according to objective criteria. With the support of Rusch (Kernen, Germany), we developed a measurement balloon for use in laparoscopic Nissen fundoplication. The balloon allows the surgeon to define the width of the wrap and predetermine its length, as well as to measure its tension. Depending on the measured balloon pressure, the surgeon can perform fundic sutures more or less tightly. On the basis of 41 fundoplication model tests, we found that a fundic wrap typically described as "loose and floppy" produced a balloon pressure of 50-60 mmHg. In 10 laparoscopic Nissen fundoplications on domestic pigs, we were able to adjust the fundic wrap intraoperatively to a balloon pressure in this range (mean; 53.5; SD; 2.25). After the optimal intraoperative balloon pressure in humans has been investigated in a prospective study that is in progress, the Tubingen balloon is expected to serve as an instrument for quality assurance in reflux surgery. PMID- 10790559 TI - Robotics and telemanipulation technologies for endoscopic surgery. A review of the ARTEMIS project. Advanced Robotic Telemanipulator for Minimally Invasive Surgery. AB - In endoscopic surgery, the ability to guide the instrument is significantly decreased compared with open surgery. Rigid laparoscopic instruments offer only four of the six degrees of freedom required for the free handling of objects in space. Robotics technology can be used to restore full mobility of the endoscopic instrument. Therefore, we designed a master-slave manipulator system (ARTEMIS) for laparoscopic surgery as a prototype. The system consists of two robotic arms holding two steerable laparoscopic instruments. These two work units are controlled from a console equipped with two master arms operated by the surgeon. The systems and its components were evaluated experimentally. Laparoscopic manipulations were feasible with the ARTEMIS system. The placement of ligatures and sutures and the handling of catheters were possible in phantom models. The surgical practicability of the system was demonstrated in animal experiments. We conclude that robotic manipulators are feasible for experimental endoscopic surgery. Their clinical application requires further technical development. PMID- 10790561 TI - The clinical suitability of laparoscopic instrumentation. A prospective clinical study of function and hygiene. AB - On the basis of experience gained from 6,000 laparoscopies (73% cholecystectomies) at the Moabit Hospital in Berlin, we carried out a cohort study to analyze the failure rate and decontamination of labeled "tracer" instruments processed in three test trays that were each subjected to 100 cycles. The majority of repairs focused on the functional parts of separable scissors and damaged or lost components. At 4%, the repair index after laparascopic use was less than that of a previously documented investigation period covering 1990 to 1996. A comparison of the costs of disposable and reusable instruments showed that reusable instruments were more cost-effective by a factor of > or =10, indicating that the price gap reported in our previous calculation for 1992 and 1994 has closed only slightly. After 100 cycles, we found traces of proteinaceous material in the eluate on every fourth instrument inspected (eight of 32); half of them (four) gave a positive reading when tested with a hemoglobin pseudoperoxidase test stick. It must be said, however, that similar residual contamination has been found on instruments used in conventional open surgery, with no indication of clinical relevance. This study was designed to examine the clinical suitability of laparoscopic instruments in terms of function and hygiene. Improvements in instrument design and cleanability must focus in particular on the reproducibility of cleaning results, because cleaning is the most important step in processing sterile supplies. As the number of minimally invasive operations has risen considerably, a mere visual check no longer meets the requirements prescribed by modern quality assurance. A multicenter study of residual proteins found on tracer instruments in all surgical fields is now in progress. PMID- 10790562 TI - A new remote-controlled endoscope positioning system for endoscopic solo surgery. The FIPS endoarm. AB - In the field of endoscopic solo surgery, the assistance received by the surgeon from ergonomical positioning devices is extremely important. They aid in both the retracting of instruments and the positioning of the endoscope. However, passive systems derived from open surgery have not proved satisfactory. Therefore, we set out to develop a remote-controlled arm capable of moving a rigid endoscope with about four degrees of freedom, while maintaining an invariant point of constraint motion coincident with the trocar puncture site through the abdominal wall. The system is driven by means of speaker-independent voice control or a finger-ring joystick clipped onto the instrument shaft close to the handle. When the joystick is used, the motion of the endoscope is controlled by the fingertip of the operating surgeon, which is inserted into the small ring of the controller in such a way as to make the motion of the fingertip correspond directly to the motion of the tip of the endoscope. A study was performed to compare the two different interfaces available for the system. With both interfaces, the guiding system allows for transparent and intuitive operation. Its set-up is easy; it is safe and reliable to use during the intervention; and it is faster than human assistance. With its improved ergonomy, this new generation of remote-controlled endoscope positioning system represents a further step toward the diffusion of solo surgery techniques in minimally invasive therapy. In our opinion, this prototype creates a valid compromise between human and robotic control of rigid endoscopes. PMID- 10790563 TI - Laparoscopic radiofrequency ablation of primary and metastatic liver tumors. Technical considerations. AB - BACKGROUND: Radiofrequency thermal ablation is a new technology for the local destruction of liver tumors. Since we first described laparoscopic radiofrequency ablation (LRFA) for the treatment of liver tumors, much has been learned about patient selection, laparoscopic ultrasound (LU) guided placement of the ablation catheter, monitoring of the ablation process, and patient follow-up. METHODS: Since January 1996 we have performed LRFA of 250 tumors in 67 patients including 85 adenocarcinomas, 107 neuroendocrine tumors, 34 sarcomas, 1 melanoma, and 11 hepatomas. We used LU to guide placement of the ablation catheter and to monitor the ablation process. Most of the patients had two trocars (camera and laparoscopic ultrasound) with the 15-gauge ablation catheter (RITA Medical Systems, Mountain View, CA, USA) placed percutaneously. RESULTS: The LRFA procedure was completed successfully in all patients, with 1 to 14 lesions per patient, ranging in size from 0.5 to 10 cm in diameter. The entire liver could be examined by LU via right subcostal ports. Criteria for successful ablation were 5 min ablation times at 100 degrees C with 1-min cool-down temperatures of 60 degrees to 70 degrees C. Outgassing of dissolved nitrogen, monitored by ultrasound, was useful in confirming the zone of ablation. Intralesional color flow Doppler, seen before ablation, was eliminated after ablation. Placement of the grounding pad closer to the lesion on the back rather than the thigh resulted in more efficient energy delivery to the tumor. Lesions larger than 3 cm in diameter required overlapping ablations to achieve a 1-cm margin of normal liver. Most patients required overnight hospitalization, with no coagulopathy or electrolyte disturbances noted. CONCLUSIONS: The LRFA procedure is a novel, minimally invasive technique for treatment of liver tumors that have failed conventional therapy. This study documents the technical aspects of targeting lesions and performing reproducible zones of ablation. Familiarity with these techniques should lead to more widespread application. PMID- 10790564 TI - Teamwork in digestive intervention. PMID- 10790565 TI - A simple technique for delivery of bulky gallbladder during laparoscopic cholecystectomy. A cautionary note. PMID- 10790566 TI - What is the appropriate mesh for laparoscopic intraperitoneal repair of abdominal wall hernia? PMID- 10790570 TI - Principles of treatment of choroidal neovascularization with photodynamic therapy in age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a common cause of legal blindness in the developed countries in people older than 50 years of age. AMD complicated by choroidal neovascular membranes (CNV) accounts for 12% of AMD, but for 88% of legal blindness cases. Because of the progressive aging of the population, it is expected that AMD will be one of the greater public health problems in ophthalmology in the first half of the next century. Laser photocoagulation remains the only proven treatment for CNV in AMD, but unfortunately, is applicable only to a minority of patients presenting with CNV in AMD. Photodynamic therapy (PDT) is a new experiment treatment for CNV that combines the application of low-intensity light with a photosensitizing agent in the presence of oxygen to produce tissue effects. It uses the noninvasive potential of the laser light to cause a nonthermal localized chemotoxic reaction and obtain highly selective occlusion of the neovascular channels, with sparing of the overlying photoreceptors. Animal studies showed that PDT accounts for the effective closure of experimentally induced CNV. Phase I-II clinical studies showed that PDT using BPD can safely stabilize leakage from CNV in a majority of patients for up to 3 months. Phase III clinical studies to assess the long term prognosis of PDT-treatment of CNV in AMD are ongoing. PMID- 10790572 TI - Laser treatment of retinal diseases by subthreshold laser effects. AB - The retinal pigment epithelium (RPE) may play an important role in the therapeutic effect of laser photocoagulation. This suggests a benefit from targeting the RPE instead of the sensory retina. Calculating the actual temperature profile after laser photocoagulation, it can be shown that pulse durations have to be in the order of microseconds (micros) to spare the retina from damage. This is consistent with histological findings after laser exposure with repetitive, 5 micros Nd:YLF laser pulses (527 nm). Therapeutic effects have been shown in patients with diabetic macular edema, drusen, and central serous retinopathy. PMID- 10790571 TI - Transpupillary thermotherapy. AB - Hyperthermia has long been recognized as potentially useful in the treatment of human neoplasms. Only recently has technology allowed hyperthermic treatment to be delivered to ocular structures in the form of ultrasound, microwave, or ferromagnetic energy. A novel technique, transpupillary thermotherapy, allows the direct application of hyperthermic energy to posterior segment ocular structures. The treatment of two posterior segment diseases, choroidal melanoma and choroidal neovascularization attributable to age related macular degeneration, are reviewed in this article. PMID- 10790573 TI - Indocyanine green angiography and retinal sensitivity after photodynamic therapy of subfoveal choroidal neovascularization. AB - Photodynamic therapy (PDT) is currently being evaluated in clinical trials for the treatment of choroidal neovascularization (CNV). Preliminary results indicate that PDT achieves immediate absence of leakage from CNV while maintaining visual acuity. Indocyanine green angiography reveals a reduction in CNV size and a persistent decrease in leakage activity after PDT. PDT appears to be characteristically accompanied by choroidal perfusion changes that regularly resolve within 3 months. Microperimetry shows an improvement of the central visual field with a decrease in scotoma size and intensity. Repeated PDT applications do not cause additional damage to the treated area, but might further enhance the recovery of macular function. A placebo-controlled, multi center trial (TAP trial) evaluating the benefit of repeated PDT treatments in 3 month intervals is currently underway. PMID- 10790574 TI - Laser photocoagulation of eyes with drusen: will It help? AB - Drusen are deposits located deep in the sensory retina that are seen characteristically in eyes with macular degeneration (AMD). As multiple large drusen are a risk factor for the development of visual loss from AMD, laser strategies to cause drusen to resorb have been devised. Two large, randomized controlled trials using laser photocoagulation prophylactically for the treatment of AMD are underway to determine the efficacy of such treatment. PMID- 10790575 TI - Prostaglandin analogs in the treatment of glaucoma. AB - Prostaglandin (PG) analogs are some of the most recent additions to the list of ocular hypotensive medications. Two analogs of naturally occurring PGs are available commercially, isopropyl unoprostone (Rescula [Ciba Vision, Atlanta, GA]) and latanoprost (Xalatan [Pharmacia & Upjohn, Bridgewater, NJ]). Presently, latanoprost 0. 005% is the only PG analog commercially available in the United States. These agents have been shown to be the most effective topical medications for reducing intraocular pressure. They have a different mechanism of action than other ocular hypotensives, and act primarily by increasing uveoscleral outflow. Because of this, PGs have a substantial additive effect when used with agents that reduce aqueous production (eg, beta blockers or carbonic anhydrase inhibitors) or that increase trabecular outflow facility (eg, pilocarpine). Local side effects include mild conjunctival hyperemia and local irritation, darkening of iris color, increased growth of eyelashes, and a possible association with cystoid macular edema or iritis in some patients with other risk factors. No systemic side effects have been proven to be caused by latanoprost. Recommended dosing is once daily at bedtime. PMID- 10790576 TI - Cataract surgery in the glaucoma patient: advances and modifications. AB - Cataract removal in the glaucoma patient is a less arduous task today than it was a decade ago, because of technological advances and modifications in surgical technique. These technological advances include excellent phacoemulsification instrumentation. Useful modifications in surgical technique include the use of combined phacotrabeculectomy with antimetabolite supplementation, when necessary. Anticipated advances in the next decade include superior viscoelastics and the widespread availability of capsular tension rings. PMID- 10790577 TI - Advances in glaucoma diagnosis and therapy for the next millennium: new drugs for trabecular and uveoscleral outflow. AB - Advances in our understanding of the physiology and molecular biology of the trabecular and uveoscleral outflow pathways of the eye will lead to the development of new approaches for glaucoma therapy. Therapies of the future will target the structures and enzymes involved in maintaining cell shape and cell cell and cell-extracellular matrix interactions. Altering the extracellular matrix in the ciliary muscle has been important in the intraocular pressure lowering effects of prostaglandins and will be developed further as an approach to enhancing outflow through the trabecular meshwork. Gene therapy may be used to enhance or suppress the endogenous targets that are ultimately responsible for the outflow enhancement triggered by these agents. PMID- 10790578 TI - Antimetabolite use in revising failing filtering blebs. AB - This article evaluates the efficacy of a subconjunctival injection of mitomycin-C (MMC) followed by needling of the scar tissue to restore bleb function. By means of preoperative ultrasonic biomicroscopy, all the eyes in this study were categorized into one of three groups: Group I (bleb present), Group II (minimal bleb), and Group III (no bleb present). This classification was carried out to try to control for differences in bleb appearance that may correlate with bleb function. PMID- 10790579 TI - Saving the nerve from glaucoma: memantine to caspaces. AB - Three recent discoveries regarding the pathophysiology of human glaucoma are that retinal ganglion cells die by apoptosis, that nitric oxide synthase levels are altered, and that glutamate is elevated in the vitreous. These findings provide encouraging new avenues for the development of neuroprotective strategies to alleviate ganglion cell loss and blindness that accompanies this disease. In this article, we discuss some of these data, as well as potential therapies that may arise from these findings. PMID- 10790580 TI - New aqueous inflow inhibitors. AB - Several new aqueous inflow inhibitors have been introduced over the past few years. These new agents include two alpha agonists, two topical carbonic anhydrase inhibitors, and a novel combination agent. All of these agent compare favorably with timolol in terms of aqueous suppression, though each has its own relative advantages and disadvantages. PMID- 10790581 TI - Optic nerve blood flow in glaucoma. AB - Vascular disorders of the anterior optic nerve, historically, have been described as potential etiologic factors in the development of glaucomatous optic neuropathy. During the past several decades, clinical and experimental evidence of the involvement of vascular aberrations as a potential causative factor or associated risk factor have increased. However, the direct evidence that optic nerve ischemia contributes to glaucomatous optic neuropathy remains limited. Several questions about our current knowledge arise. Can ischemia alone or in combination with other factors cause glaucomatous optic neuropathy? Does our current knowledge of the vascular anatomy and physiology of the optic nerve allow us to understand vascular changes observed in individuals with glaucoma? Can we rely on current measurement techniques to assess and monitor the vascular beds of the optic nerve? This article summarizes the complex nature of the microcirculation in the anterior optic nerve, discusses the possible contribution of vascular factors in the development of glaucoma optic neuropathy, and examines the measurement techniques used in current studies. PMID- 10790582 TI - Advances in molecular genetics of glaucoma: a perspective for the clinician. AB - Extraordinary progress has been made over the last 5 years in improving our understanding of the molecular causes of various glaucomas. Identification of the myocilin gene and of 5 additional genetic loci that play a role in development of primary open angle glaucoma provides us with an opportunity to reclassify this complex phenotype based on underlying cause of disease. Clinicians who obtain a detailed family history from their patients and who have referred families with glaucoma to laboratory investigators are at the forefront of this revolution in molecular genetics. As a result, all clinicians will soon be poised to profit from genetic discoveries by being able to order diagnostic tests that will identify specific genetic mutations. These mutations will provide a detailed understanding of the natural history of the patient's glaucoma, as well as information for genetic counseling. Individuals at risk in the family will be able to be identified, permitting closer follow-up and earlier institution of glaucoma therapy. A new understanding of pathophysiology is developing and will provide the basis for more rational pharmacological and gene therapy approaches in the future. The hope of primary prevention for various glaucomas, once a distant dream, may become reality within our lifetimes. PMID- 10790583 TI - Advances in optic nerve head analysis in glaucoma. AB - Assessment of the optic nerve head for structural changes is critical for the detection of glaucoma and for following progression in patients with established glaucoma. Qualitative optic nerve analysis can be obtained via methods such as disc drawings and stereo disc photography. However, these methods are limited by significant interobserver and intraobserver variability. Disc photographs can also vary greatly according to the camera angle, photographic technique, lighting, and magnification. The need for accurate, reproducible, and quantitative, cost-effective techniques of assessing the optic disc and appreciation of the limitations of subjective clinical observation stimulated the development of new technologies. PMID- 10790584 TI - Nonpenetrating filtering procedures: viscocanalostomy and collagen wick. AB - Unroofing of Schlemm's canal without entering the anterior chamber has been performed previously to lower intraocular pressure. Initial results were good, but long-term results were discouraging, and the techniques were abandoned. Recently, two variations have surfaced. Visco canalostomy is a technique devised by Robert Stegmann of South Africa. The glaucoma drainage device (Collagen Wick) was initially introduced by Koslov in Russia. These techniques are described in this article. PMID- 10790585 TI - [Heart-kidney-adrenal triangle interactions]. AB - The heart-kidney-adrenal theme at the 42(nd) Journees Internationales d'Endocrinologie Clinique - Henri-Pierre Klotz held in Paris in April 1999 was chosen for at least two reasons. First, cardiovascular signs and symptoms are part of the most important and frequent endocrinopathies whose course is often characterized by cardiovascular complications. Secondly, the treatment of hypertension, heart failure and vascular nephropathies, as well as the prevention of atherosclerosis, call for homonal treatment or therapeutic intervention with agents acting upon endocrine systems that regulate cardiac, renal and vascular functions. The endocrinology specialist should therefore have full knowledge of the latest progress in the fields of the physiological and pathophysiological function of these systems, of the cellular mode of action of the hormones targeting the vardiovascular system, their genesis in classical endocrine organs, and also their local formation and action in tissues. Finally, it is important to be aware of the new therapeutic approaches opened by these recent developments. PMID- 10790586 TI - [Synthesis and local and distant actions of vasoactive peptides]. AB - In this short review, we compare the structure, biosynthetis and processing pathways, physiological actions, receptor systems and signaling pathways of 3 vasoactive peptides: vasopressin, angiotensin II and endothelins. This comparison shows clearly that all these peptides are synthesized as preproproteins and are processed by proteolytic enzymes. They all act via different types of membrane bound G protein coupled receptors. Their actions on cardiovascular target tissues are either of an endocrine nature (vasopressin) or a paracrine nature (endothelins) or both (angiotensin II). PMID- 10790587 TI - Growth hormone and cardiac function. AB - Impaired cardiovascular function, which may reduce life expectancy, has recently been demonstrated both in GH deficiency and excess. Moreover, experimental and clinical studies support the evidence implicating GH and/or IGF-I in the regulation of heart development. The existence of a specific acromegalic cardiomyophathy characterized by myocardial hypertrophy with interstitial fibrosis, lympho-mononuclear infiltration and areas of monocyte necrosis which often result in biventricular concentric hypertrophy has been recenty demonstrated. By contrast, patients with childhood or adulthood-onset GH deficiency (GHD) present with abnormalities of structure and function of the left ventricle. In these patients, a significant increase in the vascular intima-media thickness and an increased number of atheromatous plaques have also been reported. The abnormalities of cardiovascular system could be partially reverted by suppressing GH and IGF-I levels in acromegaly or after GH remplacement therapy in GHD patients. The evidence that GH is able to increase cardiac mass suggested its use in the -treatment of chronic heart failure of diverse etiologies. GH administration was -demonstrated to induce an improvement in hemodynamics and clinical status in some patients. Although these data should be confirmed in double-blind placebo controlled studies in larger series, the promising results open new perspectives for GH treatment in humans. PMID- 10790588 TI - The growth hormone and insulin-like growth factor 1 axis in heart failure. AB - Experimental data suggests that growth hormone and IGF-1 have beneficial effects on myocardial function in animal models of heart failure. Preliminary evidence suggests an abnormality in the growth hormone-IGF-1 axis in heart failure with relative growth hormone resistance. Beneficial effects of growth hormone and IGF 1 include vasodilatation, stimulation of cardiac hypertrophy, increase in calcium sensitivity of cardiac myofilaments and prevention of apoptosis. Recently, cardiac cachexia has been shown to be a powerful negative predictive factor in heart failure. Cachectic patients have higher angiotensin II levels. In the rat there is an important interaction between the renin-angiotensin system and IGF-1. Thus, angiotensin II infusion causes weight loss in part through a catabolic effect. This effect results from increased protein degradation. Angiotensin II reduces circulating and skeletal muscle IGF-1 but increases IGF-1 and the IGF-IR expression in cardiac muscle. Preliminary data suggest a potential beneficial effect of growth hormone in heart failure. Further trials are necessary to test the potential beneficial effect of growth hormone and/or IGF-1 in heart failure. PMID- 10790589 TI - Growth hormone-releasing peptides and the cardiovascular system. AB - Growth Hormone (GH)-releasing peptides (GHRPs) and their non peptidyl analogues are synthetic molecules which exhibit strong, dosedependent and reproducible GH releasing activity but also significant PRL- and ACTH/cortisol-releasing effects. An influence of these compounds on food intake and sleep pattern has been also shown. The neuroendocrine activities of GHRPs are mediated by specific receptors subtypes that have been identified in the pituitary gland, hypothalamus and various extra-hypothalamic brain regions with (125)I-Tyr-Ala-hexarelin, an octapeptide of the GHRP family. In addition, GHRP receptors were also present in different peripheral tissues such as heart, adrenal, ovary, testis, lung and skeletal muscle, with a density significantly higher than that found in the hypothalamo-pituitary -system. A remarkable specific (125)I-Tyr-Ala-hexarelin binding was observed in the human cardiovascular system where the highest binding levels were detected in ventricles, followed by atria, aorta, coronaries, carotid, endocardium and vena cava. The binding of the radioligand to cardiac membranes was inhibited by unlabeled Tyr Ala hexare lin and hexarelin as well as by GHRP-6, GHRP-1 and GHRP-2 but not by MK-677, a non peptidyl GHRP analog. In other experiments on H9c2 myocytes, a fetal cardiomyocytes-derived cell line, specific GHRP binding was found and hexarelin showed an anti-apoptotic activity. On the other hand, in vivo studies in animals and in humans showed that GHRPs possess direct cardiotropic actions. In fact, hexarelin protects from ischemia induced myocardial damage in aged and GH deficient rats while hexarelin shows a positive inotropic effect in normal subjects as well as in patients with GH deficiency. In conclusion, GHRPs possess extra--neuroendocrine biological activity and, particularly, show direct GH-independent cardiotropic effects. PMID- 10790590 TI - [Functional compartmentation of the endocrine action of cardiac natriuretic peptides]. AB - The endocrine function of the heart is to secrete Atrial and Brain natriuretic peptides (ANP and BNP). These peptides are biologically active via particulate guanylate cyclases which generate cyclic GMP, the second intracellular messenger. A polysaccharide antagonist, HS-142-1 has been recently described by a Japanese Group. Cyclic GMP is partly secreted from the target cells into the extra cellular medium in which its accumulation is proportional to the concentration of the natriuretic peptide. Neutral Endopeptidase (NEP) is a zinc ectoenzyme involved in the catabolism of natriuretic peptides. NEP is absent in plasma but present on the surface of endothelial and smooth muscle cells. NEP is mainly expressed at the apical pole of the epithelial cells of the proximal tubule in the nephron. Chronic increase in volume and pressure within the cardiac cavities is associated with the oversecretion of natriuretic peptides. This chronic phenomenon involves the recruitment of all the cardiac myocytes to express natriuretic peptide genes. The clinical application of this hyperplasic phenomenon is congestive heart failure, in which the plasma levels of natriuretic peptides correlate with the level of the -hemodynamic stress. Therefore the plasma levels of natriuretic peptides are good pronostic markers in both experimental and human heart failure. The degree of congestive heart failure as well as the plasma levels of ANP and BNP are also -correlated with the plasma and urinary levels of cyclic GMP. The plasma level of -cyclic GMP is correlated with the endothelial concentration of cyclic GMP but not with the cyclic GMP concentration in smooth muscle cells. From these experimental data, we can conclude that plasma cyclic GMP originates from endothelial cells and is related to particulate guanylate cyclase activity. In contrast natriuretic peptides do not modulate vascular wall cyclic GMP content. The natriuretic action of ANP is probably due to the interaction of the filtered peptide with the particulate guanylate cyclase at the apical pole of the epithelial cells. The apparition of peptiduria associated with natriuresis during NEP inhibition provides evidence of the action of the peptide in the urinary compartment. It is also by a urinary pathway via the macula densa that ANP, and its potentiation by NEP inhibition, decreases renin secretion. The fact that plasma levels of ANP and plasma and urine levels of cyclic GMP correlate with the degree of salt retention in congestive heart failure, provides evidence for chronic desensitization of the system. An up-regulation of Na(+), K(+), 2Cl(-) expression associated with experimental congestive heart failure has recently been shown. Similarly, a modulation of the different sodium transporter systems along the nephron could be one of the counter-regulations leading to desensitization to natriuretic peptides. In conclusion, natriuretic peptides are true endocrine peptides, secreted by the heart, transported in the plasma, filtered by the glomeruli and active at the nephron level. The molecular effector of ANP and cyclic GMP in the epithelial cells is probably the G-kinase II, isoform phosphorylating the cystic fibrosis transmembrane conductance regulator (CFTR). The exact mechanism of desensitization remains to be elucidated. PMID- 10790591 TI - Identification and role of aldosterone receptors in the cardiovascular system. AB - The cardiovascular system is now recognized as an important mineralocorticoid target. All -components required for specific and selective aldosterone effects are present in the cardiovascular system. Mineralocorticoid receptors (MR) are expressed in the heart and large blood vessels together with the 11 B hydroxysteroid dehydrogenase type II, which ensures the enzymatic protection of MR against glucocorticoids. The recent description of local vascular and cardiac aldosterone biosynthesis strongly supports an autocrine/paracrine hormonal action. Establishment of transgenic mice models of targeted overexpression of the mineralocorticoid receptor should facilitate new insights into the molecular and cellular mechanisms of aldo-sterone actions in the cardiovascular system. PMID- 10790592 TI - Therapeutic applications of angiotensin II receptor antagonists. AB - The renin-angiotensin system (RAS) represents today one of the most strategic targets of the therapy of cardiovascular diseases. During the last 30 years a number of more or less successful approaches to inhibit the activity of the RAS have been attempted. In particular, the use of ACE-inhibitiors has led to significant improvments in the outcom/treatment of hypertension, congestive heart failure, ischemic heart disease and nephropathies. On the other hand, Ace inhibitors are not specifically targeted to RAS since they interfere with an enzyme with multiple different substrates. Furthermore, the inhibition of ACE does not prevent the formation of angiotensin II through alternative pathways, and thus the inhibition of RAS is often incomplete, especially under pathologic conditions stimulating RAS. For these reasons, the recent discovery of angiotensin II receptors antagonists, which selectively inhibit the action of angiotensin II at the level of the AT1 subtype receptor, is particularly attracting. This article reviews the background, the rationale and some of the clinical findings and potential applications with this new class of compounds. PMID- 10790593 TI - Aldosterone antagonists in hypertension and heart failure. AB - Spironolactone, a competitive aldosterone receptor antagonist (ARA), has traditionally been the treatment of first choice in idiopathic hyperaldosteronism (IHA) and for preoperative management of aldosterone producing adenoma (APA). Spironolactone is partially absorbed, is extensively metabolized mainly by the liver and its therapeutic properties are attributable to active metabolite canrenone. At therapeutic doses of 25 to 400 mg per day, spironolactone effectively controls blood pressure and hypokalemia in the majority of cases. Endocrine side effect are often associated and mainly consist of gynecomastia, decreased libido and impotence in man and menstrual irregularities in women. Canrenone and the K+ salt of canrenoate are also in clinical use: they avoid the formation of intermediate products with anti-androgenic and progestational actions, resulting in a decreased incidence of side effects. Furthermore, a relatively new selective ARA compound (eplerenone) with reduced affinity for androgen and progesterone receptors, is currently undergoing clinical trials. In essential hypertension aldosterone can contribute to hypertension and increases the incidence of myocardial hypertrophy and cardiovascular events. On the other hand, inhibition of Renin-Angiotensin-Aldosterone System (RAAS) is associated with a decrease in blood pressure, with a regression of left ventricular hypertrophy and a reduction of target organ damage. Thus, ARA have been proposed as complementary treatment associated to ACE inhibitors and angiotensin receptor antagonists. Aldosterone is also known to play an important role in pathophysiolgy of congestive heart failure (CHF). In vitro and in vivo evidences suggest that aldosterone promotes myocardial fibrosis. This effect reflects direct, extra-epithelial actions of aldosterone via cardiac MR which are counteracted by ARAs in animal models. The RAAS is chronically activated in CHF. Non potassium-sparing diuretics further stimulate the RAAS and cause hypokalemia. Thus, use of ARAs in CHF was first proposed to correct potassium and magnesium depletion. At present ARAs are indicated in the management of primary hyperaldosteronism, in oedematous conditions in patients with CHF, in cirrhosis of the liver accompanied by oedema and ascites, in essential hypertension and in hypokalemic states. Its indication as adjunctive therapy of heart failure is currently under investigation. In fact, it is well known that even high doses of ACE inhibitors may not completely suppress the RAAS; aldosterone 'escape' may occur through non angiotensin II dependent mechanisms. Addition of spironolactone to an ACE inhibitor causes marked diuresis and symptomatic improvement. During the last few years, the RALES study (Randomized Aldactone Evaluation Study) was organized to explore the efficacy of combination therapy with spironolactone and ACE inhibitor in patients with CHF, class III or IV NYHA. The study was stopped 18 months early because the results were so statistically and clinically significant that it would be unethical to continue the trial. It is reported a 30 percent decrease in mortality and hospitalisation for cardiac causes in spironolactone-treated group vs placebo group. PMID- 10790594 TI - [Experimental angiogenesis : strategy for the functional study of the transcription factors of the Ets family during morphogenesis of the vascular tree]. AB - During morphogenesis of the vascular tree, the massive outgrowth of primitive capillaries is followed by the development and the maturation of some capillary branches whereas others regress. The direct observation and the manipulation of in vivo models, including a series of recent knock-out experiments, allow to delineate the mechanisms controlling this process, and to identify factors involved in the formation of a mature capillary, surrounded with a basal lamina and pericytes. The expression of several members of the Ets family of transcription factors, Ets1, Erg and Fli, correlates with the occurrence of invasive processes, such as angiogenesis during normal and pathological development. The description of the phenotype of cultured endothelial cells expressing the DNA binding domain of Ets1 suggests that members of the Ets family take part in the morphogenesis of the -vascular tree. Although transient transfection experiments allowed the identification of putative targets genes for Ets1 during angiogenesis, deciphering the Ets1 regulation networks remains a major goal for the future. PMID- 10790595 TI - Vascular endothelial growth factors and angiogenesis. AB - VEGF was discovered in 1989. Research -conducted over the past 10 years has demonstrated that VEGF is a major regulator of angiogenesis and vasculogenesis. This paper reviews the molecular data on the multiple forms of VEGF, their signalling and accessory receptors. Five genes encoding VEGF-like proteins have been identified; the different isoforms of each VEGF molecule are generated by alternative splicing mechanisms. The different VEGF's recognize signalling tyrosine kinase receptors (Flt-1, Flk-1 and Flt-4) and accessory receptors. VEGF expression is stimulated by hypoxia-dependent and -independent mechanisms. Hypoxic responses are mediated by specific transcription factors that are expressed in a tissue dependent fashion and that are developmentally regulated. VEGF is thought to play a role in tumor angiogenesis and may contribute to cardioprotection in ischemic heart -diseases. Its role in pulmonary hypertension induced by chronic hypoxia is discussed. This review also stresses the difficulty of applying results from in vitro -studies to in vivo situations. PMID- 10790596 TI - [The pharmacology of endothelin and its antagonist bosentan]. AB - Endothelin is one of the most potent vasoconstrictors known, but it also plays a role in inflammation, fibrosis and cellular hypertrophy. Endothelin is a key pathogenic factor in congestive heart failure, essential hypertension and pulmonary hypertension. Bosentan is a specific mixed antagonist of endothelin receptors. Preclinical and clinical data suggest that bosentan might become a new approach for the chronic treatment of these cadiovascular and pulmonary diseases. PMID- 10790597 TI - [Estrogens and the arterial wall]. AB - Two isoforms of oestrogens receptor (alpha and B) have been identified in the cells of the arterial wall, and an heterogenity of their expression according to the animal species, to the vascular bed and to sex has been reported. Estrogens can thus directly influence the vascular physiology through a genomic mechanism, but extra-genomic mechanisms responsible for a short-term effect have also been suggested. Endothelium appears to be an important target for estradiol, because this hormone potentiates endothelium-dependant relaxation through an increase in NO bioavailability, and accelerates endothelial regrowth. In the model of apolipoprotein E-deficient mice, as the atrhroprotective effect deposit. The immune system appears to play a key role, as the athroprotective effect of estradiol is absent in mice deficient in T and B lymphocytes. Estrogens potentiate the endothelium-dependant relaxation through the increase in nitric oxide bioavailability. Endothelial dysfunction (abnormality of the endothelium dependent vasodilation) occurs in atheromatous arteries. Estrogens prevent and even correct this endothelial dysfunction. In monkeys, this beneficial effect of estrogens is not altered by coadministration of progesterone, but is abolished. PMID- 10790598 TI - Gene therapy for coronary disease. AB - Gene transfer offers an approach to the study and treatment of coronary disease. The localized nature of vascular diseases like restenosis has made the application of genetic material an attractive therapeutic option. Viral and non viral vectors have been developed to facilitate the entry of foreign DNA into cells. Vector improvement and production, demonstration of vector safety and therapeutic efficacy are among the main present challenges. Therapeutic angiogenesis using gene transfer is a new strategy for the treatment of coronary disease. This approach is currently being investigated in clinical trials in patients with coronary diseases. Other potential targets for genetic treatment in cardiovascular diseases include thrombosis, reendothelialization, and extracellular matrix synthesis. PMID- 10790599 TI - [Pain management (meeting organized by the 2nd section of the National Academia of Pharmacy, 16 June 1999)]. PMID- 10790601 TI - [Difficulties in child pain management]. AB - Paediatric pain has been recently recognized as a main unsolved topic. Medical knowledge provides enough data to treat most of paediatric pain. Good practices for pain control are not easy to apply because of reluctance from caregivers. Pain assessment tools are reliable but not enough used within paediatric wards. PMID- 10790600 TI - [Pain and its main transmitters]. AB - The pain message originates peripherally from a great variety of substances either released from preformed stores or extemporaneously synthetized. They stimulate or sensitize nociceptors which are associated with the peripheral endings of sensitive protoneurones. Their central endings release many types of transmitters in the dorsal horn of medulla (substance P, NO, CGRP.). At this level their release, triggered by the firing rate, is modulated by the stimulation of various presynaptic receptors operated by transmitters produced by either interneurones (enkephalins) or medullar descending neurons (dopamine, norepinephrine, serotonine). These modulations correspond to the so-called gate control. The sensitive consecutive neurones which climb towards various brain areas are submitted to contradictory influences. Several of them enhance the pain perception (nociceptin, cholecystokinin, neuropeptide FF.) whereas several other reduce it (endorphines, neurotensin, neuromedin N, anandamide.). PMID- 10790602 TI - [The hospital pharmacist and pain management]. AB - In the field of pain control, the aim of the hospital pharmacist is to establish in cooperation with clinicians the guidelines allowing an optimal pharmacological management of pain. This difficulty is increased by age or resistance to treatment condition. In such case, the hospital may encourage clinical research programs incitating to develop knowledge and extend the availability of new drugs as it is the case for opioid rotation. Hospital pharmacis are also competent in pharmacokinetics, a fruitful research objective in the field of pain control. Thus, morphine metabolite studies and efficacy versus inefficacy parameters of drug treatments are very useful for a complete understanding of opioid pharmacology. Hospitals may encourage new therapeutic approaches optimizing clinical outcomes, minimizing costs and risks. PMID- 10790603 TI - [Pain management: a priority for public health]. AB - In France, the actual anti-pain triennial plan (1998-2000) combines: the listening and the follow-up of the patient, the reinforcement of the fight against pain in health structures and care networks, the permanent information of medical and paramedical staffs, the information of the public. PMID- 10790605 TI - [Recent pharmacologic approaches to pain]. AB - The identification of compounds that can effectively and safely treat chronic pain is a major challenge of biomedical research. One approach is to optimize currently active analgesic treatments, notably by reducing their side effects. An example of this approach is the recent development of a novel class of nonsteroidal antiinflammatory drugs (NSAIDs), the cyclo-oxygenase 2 inhibitors, that lack the limiting gastrointestinal side effects of the traditional NSAIDs. Another approach, based on the recent development of molecular biology, is to develop analgesic compounds acting on new targets. These include notably ion channel blockers (TTX-resistant sodium channel blockers specific for nociceptors, N-type calcium channel blockers), nicotine receptor agonists, peptide receptor antagonists, N-methyl-D-aspartate (NMDA) receptor antagonists, vanilloids, new opioid compounds, cannabinoids, selective alpha2-adrergic agonists, purinergic modulators. Most of these compounds are currently in preclinical or early clinical investigation. However, the development of more predictable in vitro and in vivo tests and the broadening use of clinical models of experimental pain, will hopefully help increase the proportion of drugs that will become successful analgesics in clinical practice. PMID- 10790604 TI - [Pain not associated with hypernociception. A physiopathologic, clinical and therapeutic approach]. AB - It would be simplistic to consider the reality of pain as nothing more than heightened sensitivity to the pain. There are actually five types of pain, and it is often difficult to distinguish between them. Whereas hypernociceptive pain is mainly treated with analgesics belonging to one of the three categories as defined by the WHO, the mechanisms and treatment of the other four types of pain are very different It is obvious that close co-operation between the various healthcare professionals is indispensable in the treatment of these patients, while keeping in mind that the physician is "a symbol, an intermediary, a medicine that's better than medicine: the medicine works because it's a symbol - it's a symbol because it works". (P. Queneau and G. Ostermann.) PMID- 10790606 TI - [Secure prescriptions]. AB - In an attempt to facilitate access to pain relief treatments, the registered prescription sheets formerly used in France for the prescription of narcotics have been replaced by protected prescription forms complying with the technical specifications established by the AFNOR standards. When these new prescription forms were implemented, there was no modification in the legal regulations controlling narcotic supply to pharmacists. PMID- 10790607 TI - [Round table]. PMID- 10790608 TI - [ [In Process Citation] PMID- 10790609 TI - [ [In Process Citation] PMID- 10790610 TI - [ [In Process Citation] PMID- 10790611 TI - [Should we abandon internship?]. PMID- 10790612 TI - [Mechanical sutures in digestive surgery. Guidelines of the French Society of Digestive Surgery]. PMID- 10790613 TI - [Radiotherapy and chemotherapy in the treatment of cancer of the rectum]. AB - The North American consensus conference held in 1990 concluded that the best currently available adjuvant treatment for cancer of the rectum (T3, N1 to N3) was postoperative combination radiotherapy and chemotherapy. In 1994, the consensus conference held in Paris concluded that the benefit observed after preoperative irradiation warranted assessment of the effect of preoperative radiochemotherapy. To decide between these two consensus conclusions, it would be most logical to compare preoperative radiotherapy with postoperative radiochemotherapy in a group of patients with similar echo-endoscopic or imaging findings. PMID- 10790614 TI - [Surgical treatment of acute sigmoiditis]. AB - Virtually unknown at the beginning of the century, diverticulosis has become very prevalent in western countries. The natural history of the disease is notable for its acute, sometimes recurrent, attacks of diverticulitis and the significant risk of serious complications : abscess, fistula, peritonitis. CT scanning predominates in the diagnostic evaluation. It can provide both diagnostic and prognostic information. Most mild attacks of diverticulitis respond well to medical therapy while surgical treatment is indicated in the complicated forms of the disease. Surgical therapy has gradually evolved from a complicated three stage approach to two-staged, single-staged, or even minimally invasive procedures. The current trend is to intervene surgically well after the resolution of the acute attack. To achieve this, liberal use of percutaneous drainage of collections and medical treatment are advocated. The decision to perform prophylactic sigmoid resection must be based on a balance assessment of risk factors including age, severity of attacks and their recurrence. PMID- 10790615 TI - [Cephalic duodenopancreatectomy]. PMID- 10790616 TI - [Extended colpohysterectomy with pelvic lymph node dissection]. PMID- 10790617 TI - [Mobilization of the left colonic angle]. PMID- 10790618 TI - [Gastric tricobezoard]. PMID- 10790619 TI - [Prevention of anastomotic dehiscence in colorectal surgery]. AB - Anastomotic leakage is a major cause of mortality in colorectal surgery. Some variables associated with a high-risk level for anastomotic leakage have been identified, including denutrition, obesity, smoking, level of the anastomoses, qualification of the surgeon. Several methods have been evaluated in order to prevent anastomotic leakage. Prophylactic antibiotic therapy has been recommended, but has no effect on the rate of anastomotic leakage. Optimal preoperative colonic preparation appears to be obtained with povidone iodine enemas associated with a low residue diet. Stapling gives better results than sutures only for difficult anastomoses. Colostomy does not prevent leakage but minimizes its consequences. Omentoplasty, peritoneal drainage, gastric tube, biofragmentable anastomotic ring, and Coloshield have demonstrated their efficacy. PMID- 10790621 TI - [ [In Process Citation] PMID- 10790622 TI - [ [In Process Citation] PMID- 10790620 TI - [Surgery for ampulloma in a 46-year-old man]. PMID- 10790623 TI - [Gestational trophoblastic diseases. Classification, epidemiology and genetic data]. AB - Gestational trophoblastic diseases amalgamate several entities with a common denominator which is a hypersecretion of hCG: complete mole, invasive or not, partial mole or triploid syndrome, gestational trophoblastic carcinoma and trophoblastic carcinoma from the implantation site. These entities differ by their origins, their morphology, their evolution and their treatments. Complete moles are diploid and in 80% of cases, chromosomes are only from paternal origin (diandry or dispermy). Their evolution is unpredictable whatever the molecular biology or cytogenetical methods are. Partial moles, generally triploid, are much more frequent (10-20% of miscarriages) than the number of cases diagnosed during the pregnancy. In 85% of cases, two sets of chromosomes are of paternal origin. The gestational trophoblastic carcinoma is diploid and its genetic material comes from both parents. This probably excludes a direct filiation between complete mole and gestational trophoblastic carcinoma. The trophoblastic tumor from the implantation site comes from the trophoblast of the implantation site which explain why its evolution and its prognosis are totally distinct from the previous one. In this report, we successively discuss the natural history of gestational trophoblastic diseases, their epidemiology and the genetic data explaining their origins. PMID- 10790624 TI - [Heterotopic pregnancy: a report of 5 cases and review of the literature]. AB - Heterotopic pregnancy, also called combined pregnancy, associates extra-uterine pregnancy and intra-uterine pregnancy. We report five cases of heterotopic pregnancy illustrating this condition and reviewed the recent literature from 1994 to 1999. We compared the results of this review with earlier data in the literature from 1971 to 1993. We observed that therapeutic management of the extra-uterine pregnancy and course of the intra-uterine pregnancy have changed little over this period. Conversely, the frequency of heterotopic pregnancy has steadily increased since the development of medically assisted reproduction, although spontaneous cases continue to occur. The diagnosis of heterotopic pregnancy is often difficult as the symptomatology is often misleading. Transvaginal ultrasound generally gives the diagnosis which may be confirmed by laparoscopy, allowing treatment of the extra-uterine pregnancy. Maternal and intra-uterine fetal prognosis depends on early diagnosis which should be made, if possible, prior to termination of the extra-uterine pregnancy. PMID- 10790625 TI - [Stereotactic excisional nonpalpable breast lesions using the advanced breast biopsy instrumentation]. AB - Today, the widespead use of mammography has increased the detection of suspected nonpalpable lesions. Accordingly, the number of surgical biopsies is higher. PURPOSE: Evaluation of Advanced Breast Biopsy Instrumentation (ABBI) system as a promising alternative to conventional surgical biopsy in stereotactic excisional breast biopsies less than 20 mm. METHODS: From October 1998 and May 1999, 33 women have had: stereotactic excisional breast biopsies less than 20mm using ABBI. Epidemiologic, mammographic and histologic datas were collected. RESULTS: Mammographic finding were 23 microcalcifications and 10 nonpalpable solid nodular densities. In 25 cases, pathology were benign (79%), 7 cases (21%) were cancer. In these cases, the cancer could be treated later, with a single surgical procedure. No complication occurred and tolerance was excellent in most cases. CONCLUSION: Stereotactic excisional breast biopsies less than 20 mm using ABBI is a very good alternative (reliable, reproducible, ambulatory, very weak morbidity) to surgical biopsy. PMID- 10790626 TI - [Comparison of the Bologna and Ingelman-Sundberg procedures for stress incontinence associated with genital prolapse: ten-year follow-up of a prospective randomized study]. AB - OBJECTIVE: Our purpose was to evaluate and compare the long-term results of the Bologna and the Ingelman-Sundberg procedures for the treatment of stress urinary incontinence in women with genital prolapse. PATIENTS AND METHODS: Forty-seven women underwent surgery at the gynecologic division at Dunkirk Hospital, France between January 1989 and August 1990. All patients presented a genital prolapse with a cystocele of at least degree 2 associated with urinary stress incontinence. The subjects were randomly allocated to one of the two procedures. In the clinical incontinence group (28 patients), 12 procedures were Bologna operations and 16 were Ingelman-Sundberg operations. In the potential incontinence group (19 patients), 11 procedures were Bologna operations and 8 were Ingelman-Sundberg operations. Physical examination and urodynamic explorations were performed preoperatively, and 3 months and 1 year postoperatively. A questionnaire was sent to all participating women during the tenth year of follow-up. We obtained 46 answers. RESULTS: Mean follow-up was 9.7 years. The result of the 1-year postoperative evaluation has been previously published. At 1 year, complete cure was achieved in 91.7% of the patients who underwent the Bologna procedure and 93.7% in those who had the Ingelman-Sundberg procedure. At 10 years, the cure rate was 72.7% and 56.2% (p<0.05) respectively. After the first year, the decline in cure rate was twice as fast with the Ingelman-Sundberg procedure than with the Bologna operation. CONCLUSION: The longevity of the Bologna procedure is greater than that of the Ingelman-Sundberg procedure. Recovery rate declines by 20% in 9 years. All results of urinary stress incontinence surgery were good after the first year. One has to wait 5 to 10 years before reliable informative results can be obtained. This fact should be taken into consideration before accepting to use of new procedures. PMID- 10790628 TI - [Ultrasonographic diagnosis and prenatal management of fetal ovarian cysts]. AB - OBJECTIVE OF THE STUDY: To evaluate the outcome of fetal ovarian cysts in relation to their ultrasonic appearance and size. To define, on that basis, the contribution of intrauterine aspiration. MATERIAL AND METHODS: Retrospective study of all ovarian fetal cysts detected by prenatal ultrasound examination (n = 25) during a 4 year period. All these cysts were followed during pregnancy and after delivery until spontaneous or surgical resolution. RESULTS: At the time of prenatal sonographic detection at the mean gestational age of 32 1/2 weeks, the mean cyst diameter was 43+/-17 mm and 36% of all these cysts were already complicated (fluid-debris level, septa or finding of a retracting clot). Among the 16 non-complicated cysts (echolucent and thin-walled) 44% became twisted during the perinatal period irrespective of the size or the time of discovery. At birth, all these complicated cysts underwent surgical treatment and needed oophorectomy or adnexectomy. Surgery was therefore performed in a total of 56% of neonates. The pathologic reports confirm in all cases the benign follicular or follicular lutein nature of the cysts. CONCLUSION: Due to this high rate of mechanical complications, cyst decompression may be considered at the time of diagnosis in case of an anechoic fetal ovarian cyst. The safety and efficacy of this approach, on the cases reported in the literature seem encouraging. Although, a prospective randomized evaluation is needed. PMID- 10790627 TI - [Laparoscopic surgery and "massive" obesity]. AB - OBJECTIVE: Massive obesity is an important risk factor in gynecology surgery. The traumatic effect of traditional laparotomy on the parietal wall is responsible for important per and postoperative morbidity. We evaluated the feasibility and advantages of the laparoscopic approach in these patients, both in terms of surgical procedure and anesthesia. METHODS: To evaluate the technique, we reviewed four patients with massive obesity (BMI > or =40 kg/m(2)) who underwent laparoscopic surgery in our department. For each patient, we studied the cardiovascular risk factors, indications for operation, surgical technique, anesthesia conditions and follow-up. RESULTS: From the surgical point of view, certain technical difficulties were noted such as the problem of exposure and coagulation difficulties for the vascular pedicles enveloped in a layer of fatty tissue. No conversion to laparotomy was necessary. From the anesthesiology point of view, unlike what was previously feared, there was a reduction in the high operative risk due to obesity, especially due to postoperative benefits. Follow up in these four patients was uneventful. CONCLUSION: Patients who suffer massive obesity are in a high risk category for surgery and anesthesia. This high risk group can benefit most from the advantages of laparoscopic surgery compared with open surgery. PMID- 10790629 TI - [Prenatal diagnosis of congenital diaphragmatic hernia: evaluation of the prognosis]. AB - OBJECTIVE: The aim of this study was to evaluate prognostic prenatal factors of congenital diaphragmatic hernia. MATERIAL AND METHODS: We designed a retrospective study of 34 patients with congenital diaphragmatic hernia. The infants were delivered at the Edouard Herriot Hospital between September 1, 1994 and June 30, 1998. We excluded cases of pregnancy termination. After eliminating 4 cases, we studied 30. The factors studied were: polyhydramnios, transverse abdominal diameter, hepatic and umbilical vein deviation, mediastinal deviation, intrathoracic location of the stomach. Prenatal karyotype and echocardiography were systematically obtained. RESULTS: Total perinatal mortality was 53% and was 48% in case of unique diaphragmatic hernia. Prognosis was poorer if the diaphragmatic hernia was associated with another malformation (perinatal mortality: 80%, 4/5 cases), or in case of preterm delivery (83%, 5/6 cases). Factors associated with poor prognosis were: polyhydramnios, transverse abdominal diameter below the 5(th) percentile and major hepatic deviation. Diagnosis before 25 weeks was not associated with poor prognosis. Intrathoracic stomach was a good diagnostic sign, but did not allow an assessment of prognosis. CONCLUSION: Diagnosis of congenital diaphragmatic hernia was made before 25 weeks in 77% of the cases. It was not a factor of poor prognosis. As other authors, we found that prenatal association with another malformation (especially cardiac malformation) polyhydramnios, deviation of the liver, and abdominal transverse diameter below the 5(th) percentile were factors of poor prognosis. But it was difficult to determine the prenatal prognosis. Improvement is needed. PMID- 10790630 TI - [Percutaneous umbilical blood sampling in pregnant women with thrombocytopenia. Apropos of 35 cases]. AB - OBJECTIVE: To determine the interest of percutaneous umbilical blood sampling in maternal thrombocytopenia. PATIENTS AND METHOD: We performed a retrospective study of 36 percutaneous umbilical samples in pregnant women with thrombocytopenia. We included patients with a platelet count less than 100,000/mm(3) or a previous diagnosis of idiopathic thrombocytopenic purpura. Patients were divided into two groups. Group A consisted of women whose thrombocytopenia was caused by idiopathic thrombocytopenic purpura. The other patients were included in the group B. Percutaneous umbilical blood samples were performed by direct ultrasound guidance under sterile conditions, after 37 weeks gestation. Cesarean section was indicated if the fetal platelet count was < or =50 000/mm(3), in other cases obstetric indications were followed. RESULTS: Percutaneous umbilical blood samples were performed at a mean age of 38.5 gestational weeks. Fetal blood sampling was successful in 34 of the 36 cases. There was no fetal complication. In group A, two fetuses had low cord-blood platelet counts (20,000 and 98,000 per cubic millimeter). There was no fetal thrombopenia in group B. Three cesarean sections were performed, in one case because of severe fetal thrombocytopenia and in 2 cases because of an unsuccessful percutaneous umbilical blood sampling (in both, neonatal platelet count was normal). There was no significant complication in infants. There was a significant correlation between fetal and neonatal platelet count. CONCLUSION: There is no maternal predictive factor for fetal thrombocytopenia. Percutaneous umbilical blood sampling is feasible and has a good diagnostic value. The overall risk of fetal loss is low. But there is no indication of percutaneous umbilical blood sampling in the management of thrombocytopenia in pregnancy because severity of thrombopenia is not directly related to the severity of maternal disease and there are no series large enough to assess the association between delivery method and intracranial hemorrhage in thrombocytopenic infants born of mothers with idiopathic thrombocytopenic purpura. More subjectively than on scientific arguments a percutaneous umbilical blood sampling could be performed for patients with idiopathic thrombocytopenia purpura and a past history of neonatal intracranial hemorrhage. A cesarean section could be done if the platelet count is less than 100 000/mm(3), no intracranial hemorrhage has been described above this level. PMID- 10790631 TI - [Medical termination of pregnancy for fetal anomaly: the patient's point of view]. AB - OBJECTIVES: To analyze the patient's point of view concerning pregnancy termination for fetal anomaly. METHODS: A questionnaire concerning the different steps of medical termination of pregnancy was given to 103 women on day 2 after termination. RESULTS: Most women thought that they were the ones who should make the decision (67%). Complete information prior to the procedure was greatly appreciated (81%). Physical pain remained one of the main concerns for patients given Dilapan. 94% of the women had epidural anesthesia before induction. Various mourning patterns were observed. Only 41% of the women wished to see their baby after termination; there was a correlation with age of pregnancy and social environment. Psychological assistance involved the entire team and a consultation with a pedopsychiatrist (81%). The most painful moment was the moment when breaking the new of the fetal anomaly. CONCLUSION: The women were very much in need of expressing their sorrow very soon after the event. Team work and lack of rigidity in care taking enhances the expression of individual resources, both by the medical team and the patients. Three points were highlighted by the patients. -the desire to participate in the decision making;--the importance of in-depth information on technical aspects of the procedure;--initial new breaking is recognized as a major trauma. PMID- 10790632 TI - [Newborn shoulder width: physiological variations and predictive value for shoulder dystocia]. AB - OBJECTIVES: The purpose of this study was to determine maternal and infant characteristics affecting newborn shoulder width (NSW) and to evaluate the predictive value of NSW measurement in cases of shoulder dystocia. DESIGN: NSW was systematically measured at birth during a period of 18 months. SETTING: Department of Obstetrics and Gynaecology of Saint-Antoine University Hospital (Paris-France). POPULATION: A total of 2.222 NSW measurements were performed and 22 cases of true shoulder dystocia occurred during the study period. METHODS: NSW measurements were reviewed and correlated with maternal age, parity, nonpregnant weight, weight gain during pregnancy, height, race, fasting glucose and one hour glucose levels, gestational age, birthweight and sex of the neonate. A Receiver Operating Characteristics (ROC) curve was constructed to evaluate NSW as a test for predicting shoulder dystocia. RESULTS: The mean NSW was 122.06 mm +/-0.50 SD. Stepwise multiple regression showed that NSW was significantly associated with birthweight (p<0.001), parity (p = 0.04), and nonpregnant weight (p = 0.04). We estimated that the best cut-off for shoulder dystocia prediction was a NSW measurement with a low false positive rate (<10%) in association with a high sensitivity rate. Therefore, NSW measurement above or equal to 140 mm was selected. This measurement should have a low sensitivity of 27.27%, a specificity of 91.82%, a positive predictive value of 4.02%, and a predictive negative value of 99.01% for shoulder dystocia prediction. Nevertheless, birthweight above or equal to 4000 g should retrospectively have a better predictive value for shoulder dystocia. CONCLUSIONS: NSW measurement, which is strongly correlated with birth weight, still remains a poor predictor for shoulder dystocia, even when this evaluation is correct antenatally. PMID- 10790633 TI - [The vaginal plastron for cure of cystocele]. AB - We describe a new surgical technique (the vaginal plastron) for the treatment of cystocele by the vaginal route. The technique is based on bladder support by a vaginal strip, isolated from the anterior colpocele, left attached to the bladder, associated with a suspension of this strip by its fixation to the tendinous arch of the pelvic fascia by six lateral sutures (3 on each side of the plastron). Proposed as a curative treatment of cystocele and combined with the Richter fixation, the plastron provides a surgical solution to the problem of cystocele relapse arising after vaginal treatment of prolapse by sacro-spino fixation alone (10 to 20% according to Richter). PMID- 10790635 TI - [ [In Process Citation] PMID- 10790634 TI - [Opioid spinal anesthesia at the end of labor for potentially difficult deliveries]. AB - We report two cases of difficult delivery, one vaginal breech presentation and one vaginal twin delivery in agitated parturients who were not given epidura analgesia. Maternal agitation was caused by pain and led to the risk of difficult fetal extraction. Epidural analgesia is not useful during the second stage of labor because of delayed onset of action of the administered drugs. Spinal injection of 10 microg of sufentanil was followed by rapid analgesia, maternal sedation and atraumatic deliveries. The usefulness of this technique is discussed for analgesia during the second stage of labor when epidural analgesia has not been performed. PMID- 10790636 TI - [In Process Citation] PMID- 10790637 TI - [ [In Process Citation] PMID- 10790638 TI - [Neurological outcome of 152 surgical patients with spinal metastasis]. AB - BACKGROUND AND PURPOSE: The aim of this study is to evaluate retrospectively the benefit of the surgical spinal decompression in 152 patients with spinal metastasis. METHODS: Based on clinical notes and GP inquiry, we determined the actuarial survival curve, and assessed the pain level one month post-operatively and the motor, the sensory and sphincter distrurbances three months after surgery. RESULTS: Thoraco-lumbar lesions were usually treated using a posterior approach, with a laminectomy and if necessary an osteosynthesis. Cervical lesions were treated with an anterior approach, i.e. a corporectomy and a methylmetacrylate stabilization. Sixty eight percent of patients (103/152) had pre or postoperative radiotherapy. After the surgical decompression for a spinal metastasis, our study demonstrated an improvement in sensory status (31 % of the patients), in motor ability (56 %), in sphincter function (51 %), and a decrease in the pain intensity in 47 % of the patients. Among 83 patients who could not walk on admission (grade A, B and C of Frankel), 52 % recovered a gait function 3 months post operatively. The best benefit after surgery concerned grade C patients, of which 71 % recovered the gait function. Two percent of the patients had postoperative worsening of their motor strength. No operative mortality was noted, and the postoperative mortality rate was 3 % at 7 days and 9 % at 30 days. The analysis of the actuarial survival curve demonstrated a mean follow up of 3.7 years. The mean survival time was 12 months with 25 % of patients surviving 2 years. CONCLUSION: Surgical decompression is effective in relieving neurological symptoms from spinal metastasis. In our experience a complete motor deficit does not seem to be a good surgical indication because of the lack of postoperative improvement. PMID- 10790639 TI - [Practical role of functional MRI in neurosurgery]. AB - INTRODUCTION: Since its description in the early 1990's, functional magnetic resonance imaging (fMRI) has been used first for research purposes, and after in clinical applications in the field of neurosurgery. The purpose of this article was to critically review the literature on fMRI to achieve a better understanding of the usefulness of fMRI in brain surgery for tumors, epilepsy surgery, and radiosurgery. MATERIAL AND METHODS: Medical literature databases containing peer review articles dating from 1990 were queried for key words related to fMRI. RESULTS: On 3,065 references scanned for relevance, 256 articles with direct or indirect relationships between fMRI and neurosurgery were analyzed. DISCUSSION: fMRI can be useful in the selection of patients for whom a surgical resection is attempted and could aid in the decision-making whether to operate on a patient who has been previously considered inoperable. fMRI is a useful tool in the decisional scheme of treatment of low-grade astrocytomas or arteriovenous malformations in the rolandic area in intact or slightly impaired patients. fMRI can be repeated in selected patients with slow growing brain tumors or congenital lesions such as AVM to study cortical reorganization phenomena. In epilepsy surgery, the Wada-test could be substituted by fMRI to determine the hemispheric dominance of language. The choice of standardized tasks and a better understanding of analysis problems in the treatment of fMRI images must be achieved before drawing more accurate conclusions on fMRI and brain tumors. PMID- 10790640 TI - [Radiotherapy of high grade glioma: use of fast neutrons, therapy and enhancement by neutron capture]. AB - Among high linear energy transfer (LET) irradiations techniques, those using fast neutrons are able to eradicate glioblastoma cells. At least a 13 grays (Gy) irradiation dose has to be used, but high morbidity is observed in case of over 11 Gy irradiation. So, no therapeutic windows have been found despite the fact that more than 900 patients were included in clinical trials. Boron neutron capture therapy (BNCT) uses alpha emitting nuclear reactions, produced within tumoral cells by boron neutron captures. (10)B is specifically loaded inside tumoral cells via a boronated molecule, and the tissues are then irradiated with thermal or epithermal neutrons. Although this type of irradiation is yet considered as a regular method in Japan, USA and Europe have started clinical trials, currently in progress, in order to define the BNCT place in the post operative care of high grade glioma. Non-removable tumors may benefit from boron neutron capture enhancement of fast neutron irradiation, i.e. the combination of these two methods. Preliminary studies show that a "biological" dose enhancement of 20 % could be obtained within the tumor when a concentration of 100 microg/g of (10)B is targeted into it. These concentrations are achievable by intra arterial administration of (10)boronophenylalanine (BPA) or borosulfhydryl (BSH). Recently, some publications have also demonstrated that the thermal neutron flux yielded within the irradiated tissues could be increased. Clinical trials, using this technique, are planned in USA and Europe. PMID- 10790641 TI - [Suprasellar arachnoid cyst associated with syringomyelia. Case report]. AB - We present a case of suprasellar arachnoid cyst which was revealed by visual impairement and hypopituitarism. Neuroradiological imaging showed the peculiar association of the suprasellar cyst with cerebellar tonsillar herniation and a large asymptomatic cervical syringomyelic cavity. Surgical treatment of the suprasellar cyst allowed the reduction of both the cyst and the syrinx. A common pathophysiological mechanism of these lesions is discussed. We suggest the possibility that an initial obstruction of the basal cisterns caused the suprasellar cyst formation which led to medullar cavity formation. PMID- 10790642 TI - [Dermoid cyst of the lateral ventricle associated with ethmoidal dermal sinus. Report of a case]. AB - We report a case of a ventricular dermoid cyst associated with a dermal sinus connected with the ethmoidal cells in a patient who developed rapid symptoms of raised intracranial pressure. Computed tomography showed a cystic mass in the right lateral ventricle with a hydrolipidic image in the left frontal horn of the ventricle and associated hydrocephalus. Magnetic resonance imaging showed a heterogeneous T1 hyperintense mass with a fistulous tract communicating with the ethmoid cells. A cerebrospinal fluid ventriculo-peritoneal shunt was initially established, which required further revision. A right sided transventricular approach was undertaken in a second stage, allowing resection of a dermoid cyst. Obliteration of the dermal sinus tract was obtained using pericranial duraplasty. Clinical and imaging features are discussed. The need for total resection including the tumor capsule and occlusion of the fistula are emphasized if recurrence and infection are to be prevented. PMID- 10790643 TI - [Solitary plasmacytoma of the cranial vault. Case report and review of the literature]. AB - We report a case of solitary plasmacytoma of the cranial vault. A 58-year-old man presented with a frontal soft tissue mass. X-Ray of the skull showed a lytic lesion of the frontal bone. CT scan showed the lesion extending intra and extracranially and cerebral angiography allowed embolization of afferent arteries. Complete removal of the lesion was performed without additional radiotherapy. Two years after surgery the patient is alive, and asymptomatic. Until 1997, 35 cases of solitary plasmacytoma of the cranial vault are reported, of which only five had frontal localisation. Solitary plasmacytoma of the cranial vault has a good outcome but progression towards a multiple myeloma is possible and deserves clinical and biological follow-up. PMID- 10790644 TI - [Vertebral osteomyelitis with epidural abscess in a child with sickle cell disease]. AB - A case of paraplegia due to a thoracic epidural abscess in a 6-year-old black girl with sickle cell anemia is reported. MRI and CT scan showed abnormalities involving T6 vertebra which were consistent with osteomyelitis. A laminotomy, associated to antibiotic administration, permitted rapid neurological improvement. Although cultures were negative, the infection was probably due to salmonellae. Salmonellosis is a well-known complication in children with sickle cell disease but spinal localization is unusual. Its pathogenesis is unclear but immunological abnormalities associated to vascular obstructions by abnormal blood red cells have been advocated. The underlying etiology of vertebral abnormalities in these children is difficult to determine, but early diagnosis with modern investigations, as CT scan and MRI, is crucial to promptly begin therapy for osteomyelitis, minimizing the risk of spinal cord compression. PMID- 10790645 TI - [Sphenotemporal aneurysmal bone cyst. A new case and review of the literature]. AB - Aneurysmal bone cysts (ABC) are benign tumors of the bones, formed by multiple cysts filled with blood and eroding the bone. They are rarely located at the skull. We report a case of a right temporal ABC in a 5-year-old girl. On admission, the physical examination showed a tough and fixed tumor, with no inflammatory signs nor vascular characteristics. The neurological examination was normal. Imaging studies showed an heterogeneous extracerebral mass, eroding the temporal bone and the greater wing of the sphenoid bone. The treatment consisted in an extensive tumor resection, associated with reconstructive surgery. The outcome was uneventful (follow-up 27 months). We underline the pathogenetic, diagnostic and therapeutic features of this condition. PMID- 10790646 TI - [Anterior and antero-lateral surgery for degenerative cervical spine diseases. Role of graft and osteosynthesis. Analysis of practices of the French-speaking european neurosurgeons]. AB - The neurological complications of cervical spondylosis depend on the direct or indirect effects of the mechanical compression of the spinal cord and roots. Nobody questions the interest of a surgical decompression, when the medical treatment is ineffective, but there is no consensus on the choice of the surgical approach and on the necessity of performing a graft with or without osteosynthesis after an anterior decompression. On the initiative of the French Speaking Neurosurgical Society (SNLF), we have analyzed the practices of its European members in 1998 and report here the results of this study which collects 3 645 surgical procedures for degenerative pathology of the lower cervical spine responsible for radiculopathy or myelopathy. An anterolateral approach was used in 85.3 % of all cases, among which 87.9 % of extensive discectomy (Smith Robinson or Cloward technique) and 12.1 % of median somatotomy. In case of discectomy, no grafting was performed in 34.7 % of cases, grafting without fixation in 25.7 %, grafting with plating in 14.9 %, interbody cages in 21.9 %, other techniques in 2.8 %. In case of somatotomy, no grafting was performed in 17.8 % of cases, grafting without fixation in 15.5 %, and grafting with plating in all other cases. When a graft was performed, an autograft was used in 59.5 % of cases, hydroxyapatite in 24.4 %, and many other bone substitutes in rare cases every time. For the fixation, monocortical screwing was used in 75 % of all cases, and bicortical in 25 %. The reasons for these practices are equivocal and not based on scientific attitude. The reasons given are : to avoid discal plucking, to prevent or treat kyphosis and postoperative spinal instability, to prevent cervical postoperative pain, to help eliminate osteophytes and hypertrophy of the ligamentum flavum. PMID- 10790647 TI - Alternatives to Autogenous Bone Graft: Efficacy and Indications. AB - Bone grafting is frequently used to augment bone healing with the numerous approaches to reconstructing or replacing skeletal defects. Autologous cancellous bone graft remains the most effective grafting material because it provides the three elements required for bone regeneration: osteoconduction, osteoinduction, and osteogenic cells. Autologous cortical bone graft provides these three components to a limited extent as well and also provides the structural integrity important in reconstruction of larger defects. However, because autogenous grafting is associated with several shortcomings and complications, including limited quantities of bone for harvest and donor-site morbidity, alternatives have been used in a wide range of orthopaedic pathologic conditions. Grafting substitutes currently available include cancellous and cortical allograft bone, ceramics, demineralized bone matrix, bone marrow, and composite grafts. No single alternative graft material provides all three components for bone regeneration. The clinical applications for each type of material are dictated by its particular structural and biochemical properties. Composite grafts consisting of several materials are often used to maximize bone healing, especially where the grafting site is compromised. PMID- 10790648 TI - Isolated Medial Collateral Ligament Injuries in the Knee. AB - The management of ligament injuries in the knee has progressed significantly over the past two decades as a result of both laboratory and clinical studies that better define the healing capacity of these supporting structures. The intracapsular ligaments (the anterior and posterior cruciate ligaments) appear to have limited potential for spontaneous healing and frequently require surgical reconstruction. The extracapsular ligaments (the medial and lateral collateral ligaments), however, appear to have a fairly robust potential for healing. As a result, the need for surgical intervention is limited to specific clinical situations. PMID- 10790650 TI - Scapular Fractures and Dislocations: Diagnosis and Treatment. AB - Traumatic injuries of the scapula and the scapulothoracic articulation have received little attention in the literature, since they are uncommon. Scapular fractures constitute only 1% of fractures in general, and scapulothoracic dissociations and dislocations are extremely rare. The vast majority can and should be managed nonoperatively. However, recent experience has shown that injuries that involve significant displacement can have long-term adverse functional consequences for both the shoulder complex and the upper extremity as a whole. In these situations, surgery should at least be considered. Various scapular fractures and dislocations are discussed, with particular emphasis on those requiring operative care. PMID- 10790649 TI - Revision Total Hip Arthroplasty: The Acetabular Component. AB - Intermediate and long-term results of revision total hip arthroplasty performed with the use of a cemented acetabular component have been disappointing, with high rates of radiographic and clinical failure. Other methods of acetabular revision involving the use of threaded cups and bipolar implants have also met with high failure rates. Although the long-term results of revision arthroplasty with uncemented acetabular components, especially in terms of polyethylene wear and pelvic osteolysis, are not yet available, the intermediate results have been excellent. PMID- 10790651 TI - Disorders of the First Metatarsophalangeal Joint. AB - The two most common disorders of the first metatarsophalangeal (MTP) joint are hallux valgus and hallux rigidus. The hallux valgus deformity has been the subject of numerous clinical studies in the past decade. This information has enabled the creation of an algorithm to assist the clinician in evaluating the patient with hallux valgus and selecting the appropriate surgical procedure. The technical aspects of various operative procedures and the most common complications are reviewed. The other major disorder of the first MTP joint is arthrosis, which results in hallux rigidus. As the arthrosis progresses, there is often proliferation of bone on the dorsal aspect of the metatarsal head, which results in impingement of the proximal phalanx during dorsiflexion. The impingement causes jamming, instead of gliding, of the proximal phalanx on the metatarsal head, which results in pain. The treatment for this condition consists of debridement of the MTP joint to relieve the dorsal impingement and, in most cases, the pain. If the arthrosis is advanced in an active individual, arthrodesis is indicated. PMID- 10790652 TI - Flexor Tendon Injuries: I. Foundations of Treatment. AB - During the past 20 years, the difficult process of reestablishing satisfactory function after primary repair of flexor tendons has evolved from scientifically unsupported trial-and-error efforts to protocols based on sound laboratory and clinical investigations. Enhanced appreciation of tendon structure, nutrition, and biomechanical properties and investigation of factors involved in tendon healing and adhesion formation have had significant clinical applications. In particular, it has been found that repaired tendons subjected to early motion stress will increase in strength more rapidly and develop fewer adhesions than immobilized repairs. As a result, new and stronger tendon repair techniques have evolved, permitting the application of early passive and even light active forces. The author reviews the most recent and clinically pertinent research in flexor tendon surgery and discusses repair techniques and rehabilitation protocols based on the information provided by these studies. PMID- 10790653 TI - Flexor Tendon Injuries: II. Operative Technique. AB - The repair of flexor tendons (zones I and II) is a technique-intensive surgical undertaking. It requires a strong understanding of the anatomy of the tendon sheath and the normal relationship between the pulleys and the flexor digitorum superficialis and flexor digitorum profundus tendons in the digit. Meticulous exposure, careful tendon retrieval, and atraumatic repair are extremely important, and the repair should be of sufficient strength to resist gapping and permit the early postrepair application of motion forces. Whenever possible, the tendon sheath should be preserved or repaired, and a smooth gliding surface should be reestablished. The author describes an effective method of tendon retrieval and a simplified technique for a four-strand tendon repair with a supplementary peripheral running-lock suture. The repair is considered to maintain sufficient strength throughout healing to allow a postrepair rehabilitation protocol that will impart passive and modest active stress forces to the repaired tendons. Complications include tendon rupture, digital joint flexion contractures, and adhesions that restrict tendon gliding and ultimately necessitate tenolysis. PMID- 10790654 TI - Physeal Fractures About the Knee. AB - The knee is the most common site of injury in childhood sports, and with increased participation in organized sports, the potential for knee injuries has accordingly increased. The epiphyses and apophyses provide a site of injury unique to the immature patient. The distal femoral and proximal tibial physes and the tubercle apophysis respond differently to acute and repetitive load and often provide less resistance to traumatic forces than do surrounding ligament and bone. Treatment of displaced physeal fractures about the knee remains one of the more difficult problems in orthopaedics. Even with appropriate conservative or surgical treatment, a successful outcome is not ensured. The Salter-Harris classification system provides general guidelines regarding the risk of growth disturbance, but there are no clinical methods for quantifying the true extent of physeal damage in an acute injury. Ultimately, the value of a treatment method must be evaluated on the basis of not only the restoration of articular congruity and physeal anatomy but also the restoration of physeal function, as evidenced by the continuation of normal growth. The mechanism of injury, clinical evaluation, treatment, and outcomes for all epiphyseal injuries about the knee are discussed, as well as differences from adult injuries. PMID- 10790655 TI - Fractures and Dislocations of the Forefoot: Operative and Nonoperative Treatment. AB - Effective treatment of common bone injuries of the forefoot is dependent on a clear understanding of both the osseous anatomy of the foot and the biomechanics of gait. Obtaining a thorough history and performing a careful physical examination are especially important because the complex anatomy of the region often makes radiographic diagnosis difficult. The keys to making the correct diagnosis in the injured forefoot are detailed, with emphasis on obtaining the appropriate radiographic studies. Included in the discussion are injuries to Lisfranc's joint and the metatarsophalangeal and sesamoid joints, as well as metatarsal and phalangeal fractures. Guidelines for operative and nonoperative management of these injuries are presented. PMID- 10790656 TI - Revision Total Hip Arthroplasty: The Femoral Component. AB - The initial results with cemented femoral revision stems were disappointing, with high early loosening rates. The application of second-generation cementing techniques improved results markedly, with loosening rates of 10% at 10 years in a number of series. Bone quality and patient age also appear to be important factors in predicting the success with a cemented revision stem. The use of a long stem is not necessary to obtain these improved results. On the basis of early reports, a recently described technique in which a revision stem is cemented into impacted cancellous allograft appears promising. Results with proximally coated uncemented revision stems have been variable, with failure rates of 4% to 10% reported at only 2 to 4 years. Initial stability must be obtained if proximal coating is to be utilized. Extensively coated revision stems provide initial stability through an interference fit between the porous coating and the diaphysis. With this technique, 10-year survival rates of 90% have been achieved, and the clinical results appear to be equivalent to those obtained with cemented revision stems and modern cementing techniques. Diaphyseal fixation has also been achieved without porous coating, either with modular revision stems or with long, textured titanium stems. Isolated early reports with such stems have been promising, but 10-year results are not yet available. PMID- 10790657 TI - Tibial Plateau Fractures: Evaluation and Treatment. AB - The goal of tibial plateau fracture management is a stable, well-aligned, congruent joint, with a painless range of motion and function. Minimally displaced stable fractures should be treated with protected mobilization. The treatment of displaced tibial plateau fractures, however, remains controversial. Surgical reduction and stabilization of displaced tibial plateau fractures, when indicated, requires careful evaluation of both the "personality" of the fracture and the soft-tissue envelope. The timing of surgery and the handling of the soft tissue in this region are critical to treatment success. After restoration of a congruent joint surface, bone grafting and buttress plating are usually needed to allow early range of motion and optimize treatment outcome. PMID- 10790658 TI - The Distal Radioulnar Joint: Problems and Solutions. AB - Disorders of the distal radioulnar joint are a major source of ulnar-sided wrist pain. Fortunately, our understanding of the anatomy, joint mechanics, and pathophysiology of this area has increased greatly in recent years, making resolution of many of these problems feasible. In most cases, an accurate diagnosis can be made, and successful treatment can then be prescribed. This review covers various problems affecting the distal radioulnar joint, including fractures and dislocations, triangular fibrocartilage pathology, arthritis, and other disorders. PMID- 10790659 TI - Fractures of the Proximal Fifth Metatarsal: Selecting the Best Treatment Option. AB - Because of circulatory differences in the three zones of the proximal fifth metatarsal, the location of a fracture must be considered when selecting treatment. The most proximal portion of the base of the fifth metatarsal has good blood supply. Fractures in this zone usually extend into the fifth metatarsocuboid joint. The second zone is associated with Sir Robert Jones, who in 1902 first asserted that fractures of the fifth metatarsal are commonly caused by indirect violence. Fractures in this zone take longer to heal than more proximal fractures, and treatment should be individualized. Whether to use a functional metatarsal brace, a stiff-soled shoe, a short-leg cast, or even internal fixation with a screw depends on the patient's lifestyle and desired activity level. Fractures in the third zone occur between the distalmost portion of the metaphysis and the proximal 1.5 cm of the diaphyseal tubular bone. This zone begins just distal to the ligamentous complex holding the proximal fourth and fifth metatarsals together. In active athletes, fractures in this zone often are stress injuries. For anatomic and mechanical reasons, such fractures are the most difficult to heal. Without surgical treatment, they may take 2 to 21 months to unite and are therefore more likely to need aggressive treatment. PMID- 10790660 TI - Hip Arthroscopy: Applications and Technique. AB - Hip arthroscopy is infrequently performed in North America. The anatomic constraints of the joint and the lack of equipment specifically designed for this application have contributed to the lack of surgical experience. Because of the potential for significant neurovascular injury, familiarization with precise portal placement is essential. In properly selected patients, hip arthroscopy allows diagnosis of a variety of disorders. The authors believe this technique has a significant role in the treatment of acetabular labral tears, loose bodies, chondral injuries to the joint, and septic arthritis. PMID- 10790661 TI - Lumbar Spine Fusion in the Treatment of Degenerative Conditions: Current Indications and Recommendations. AB - The role of arthrodesis in the treatment of degenerative disorders of the lumbar spine is controversial. Most patients with these conditions can be successfully treated nonoperatively. Lumbar fusion, with or without instrumentation, is associated with more complications, especially in the elderly population. Therefore, the potential benefits to be obtained by means of arthrodesis must be measured against the risks. Arthrodesis is indicated as an adjunct to decompression for patients with spinal stenosis associated with degenerative or iatrogenic spondylolisthesis and in the treatment of progressive degenerative lumbar scoliosis and iatrogenic instability resulting from extensive decompression. The occurrence of two or more episodes of disk herniation at the same segment is a relative indication for arthrodesis. In patients with incapacitating nonradicular back pain, arthrodesis should be a consideration only after failure of a trial of nonoperative treatment lasting more than 12 months and after secondary gain issues (e.g., workmen's compensation) have been adequately resolved. Arthrodesis has a poor success rate when used to treat back pain associated with multilevel disk degeneration seen on magnetic resonance images. PMID- 10790662 TI - The Foot in Running. AB - Injuries to the foot and ankle are often encountered in runners, be they highlevel competitors or recreational joggers. Many of these injuries are due to overuse syndromes and training errors; others are related to the running surface or the athlete's footwear. Only with a rational approach to diagnosis can the primary underlying cause be identified so that appropriate treatment can be prescribed. Conservative measures, which include rest, cross-training, orthotic changes, and altering training methods, are often curative. Surgery is usually indicated only after conservative measures have been exhausted. Careful preoperative planning is needed to minimize dissection, thereby optimizing the chance of a return to the preinjury activity level. Preoperative counseling of the patient is also important, so that expectations about the outcome and the rehabilitation requirements are realistic. PMID- 10790663 TI - Anterior Cruciate Ligament Injury in the Skeletally Immature Patient: Diagnosis and Treatment. AB - Significant intra-articular knee injuries, including tears of the anterior cruciate ligament (ACL), are now being recognized more frequently in skeletally immature patients. Previously reported data on ACL tears in this age group are sparse, and studies have been of limited quality. Improvements in diagnostic techniques (e.g., physical examination signs, arthrometric testing, magnetic resonance imaging, and arthroscopy) have facilitated identification of such injuries. Hemarthrosis must be regarded as a herald of a major intra-articular injury. Surgical reconstruction options vary according to the specific diagnosis and the stage of maturity, and the available options for the very skeletally immature patient are limited. Therefore, treatment must be predicated on assessment of maturity, as determined on the basis of chronologic, radiologic, and physiologic criteria. An ACL injury in this age group is not a surgical emergency; therefore, time for discussion with the patient and his or her parents is available, so that all appropriate options can be considered. PMID- 10790664 TI - Shoulder Injuries in the Throwing Athlete. AB - The throwing athlete with shoulder pain presents a diagnostic and treatment challenge to the orthopaedic surgeon. Because pitching a baseball requires the arm to accelerate at 7,000 degrees per second, tremendous forces are experienced at the shoulder joint. Electromyographic studies have shown that the larger scapular and trunk muscles are primarily responsible for arm acceleration. The smaller and more fragile rotator cuff muscles play a significant role in decelerating the arm. During the entire throwing mechanism, the rotator cuff and the capsulolabral complex act to stabilize the humeral head on the glenoid fossa. As a result, the labrum, the capsule, and the rotator cuff are frequently the site of shoulder injury in throwers. The diagnosis of injury to these structures is based on the findings from the history, physical examination, and imaging studies. The majority of throwing injuries respond well to a carefully designed rehabilitation program. Athletes who do not improve within 6 months are candidates for surgical repair. The procedure is planned so as to minimize the amount of surgical trauma and thereby to facilitate an early return to sport. Arthroscopy is a valuable first step to confirm the pathologic diagnosis. The arthroscope alone is used to perform subacromial debridement, labral repair, or debridement of undersurface partial-thickness rotator cuff tears. If the athlete has clinical evidence of shoulder instability and arthroscopic evidence of capsular stretch, an open stabilization procedure is performed. PMID- 10790665 TI - Disorders of the Lesser Metatarsophalangeal Joints. AB - Pain in the region of the lesser metatarsophalangeal joints (often termed metatarsalgia) is a common complaint. It can be due to a variety of causes, and accurate diagnosis is essential for effective treatment. Understanding the anatomy and functions of the extrinsic and intrinsic musculature and the plantar plate, ligaments, and fat pad is important in evaluating metatarsophalangeal joint disorders. Claw toe is a hyperextension deformity of the metatarsophalangeal joint in combination with a hammer toe. Pathologic changes involving an isolated metatarsophalangeal joint may be due to monarticular synovitis. Systemic inflammatory disorders can cause variable degrees of instability, resulting in sub-luxation or dislocation. Other specific disorders at the lesser metatarsophalangeal joints include discrete and diffuse intractable plantar keratoses, Freiberg's infraction, and cock-up fifth toe. Once the specific pathologic entity has been determined, the appropriate course of nonsurgical or, if necessary, operative treatment can be instituted. PMID- 10790666 TI - Skeletal Dysplasias: An Approach to Diagnosis. AB - Skeletal dysplasias are the result of aberration in the growth and development of the skeleton. While they are individually rare, they are important in that they provide an insight into the mechanism of skeletal development. This article offers an approach to the diagnosis of skeletal dysplasias, rather than an exhaustive account of all the possible diagnoses. Dysplastic conditions are suspected on the basis of abnormal stature, disproportion, dysmorphism, or deformity. Diagnosis requires simple measurement of height and calculation of proportionality, combined with a complete physical examination, appropriate radiographs, an investigation of the family pedigree, and occasionally laboratory studies. An accurate diagnosis can usually be made on the basis of these data and a review of descriptive sources. A definitive diagnosis allows the treating physician to project the patient's ultimate height and to prognosticate about likely deformities and the risk of the recurrence of the condition in the family. PMID- 10790667 TI - Flatfoot in the Adult. AB - Flatfoot in the adult has long been a poorly understood "wastebasket" diagnosis, often used to unfairly deny asymptomatic individuals equal employment opportunities in our society. Now that flatfoot has been classified into a variety of congenital and acquired conditions, the parameters for assessment have been well defined, and rational treatment protocols have been established. Clearly, if the foot painlessly supinates/inverts to become a rigid lever for push-off and pronates/everts to absorb stress during stance, then it "functions normally" no matter what the height of the arch. However, the biomechanically offset position of pes planus with excessive heel valgus coupled with rigidity or instability can alter the connected interplay of the bones of the foot and weaken the entire kinetic chain of the lower extremity. Careful clinical and radiographic evaluation, coupled with a thorough understanding of the anatomy and biomechanics of the foot, will allow accurate evaluation and appropriate treatment. PMID- 10790668 TI - Coxa Saltans: The Snapping Hip Revisited. AB - Coxa saltans, or "snapping hip," has several causes. These can be divided into three types: external, internal, and intra-articular. Snapping of the external type occurs when a thickened area of the posterior iliotibial band or the leading anterior edge of the gluteus maximus snaps forward over the greater trochanter with flexion of the hip. The internal type has a similar mechanism except that it is the musculotendinous iliopsoas that snaps over structures deep to it (usually the femoral head and the anterior capsule of the hip). Intra-articular snapping is due to lesions in the joint itself. Diagnosis of the external and internal types is usually made clinically. Radiography can be useful in confirming the diagnosis, particularly when bursography shows the iliopsoas tendon snapping with hip motion. Other radiologic modalities, such as computed tomography, magnetic resonance imaging, and arthrography, may also be helpful, especially when there is an intra-articular cause. Most cases of snapping hip are asymptomatic and can be treated conservatively. However, if the snapping becomes symptomatic, surgery may be necessary. There may also be a role for arthroscopy in the treatment of intra-articular lesions. PMID- 10790669 TI - Running Injuries to the Knee. AB - Approximately one third of serious runners will incur an injury in a given year, and approximately one third of the injuries will involve the knee. Biomechanical studies on running reveal the tremendous cyclic forces to which the knee is subjected. There are several etiologic factors involved, including training errors, anatomic and biomechanical variations, and differences in shoes and running surfaces. Training errors, particularly rapid transitions in training, are responsible for two thirds of injuries. Knee injuries in runners must be approached with a thorough review of the training program and a complete examination of the lower extremity. Several conditions can cause anterior knee pain commonly involving the extensor mechanism, and these must be differentiated. Most knee injuries can be resolved with conservative treatment, but occasionally surgery is indicated. After a layoff due to injury or surgery, appropriate rehabilitation, including a graduated return to running, is essential to avoid reinjury. PMID- 10790670 TI - Scapular Winging. AB - Scapular winging, one of the more common scapulothoracic disorders, is caused by a number of pathologic conditions. It can be classified as primary, secondary, or voluntary. Primary scapular winging may be due to neurologic injury, pathologic changes in the bone, or periscapular soft-tissue abnormalities. Secondary scapular winging occurs as a result of glenohumeral and subacromial conditions and resolves after the primary pathologic condition has been addressed. Voluntary scapular winging is not caused by an anatomic disorder and may be associated with underlying psychological issues. The evaluation and treatment of these three types are discussed. PMID- 10790671 TI - Genu Varum in Children: Diagnosis and Treatment. AB - Genu varum is a relatively common finding in children. Physiologic bowing, which is seen most often, has a well-documented favorable natural history. Idiopathic tibia vara is the most common of the pathologic conditions that are associated with bowed legs; treatment strategies vary with the patient's age and the stage of disease and deformity. Genu varum may also accompany systemic conditions, such as achondroplasia, vitamin D-resistant rickets, renal osteodystrophy, and osteogenesis imperfecta-all of which can result in short stature. Indications for intervention are not always well defined. A rare disorder, focal fibrocartilaginous dysplasia, usually requires no treatment. Standing radiographs of the entire lower limbs are necessary for surgical planning, as the deformity can sometimes affect the distal femur rather than the proximal tibia. Restoration of the mechanical axis of the limb is the principal goal of treatment; the particular type of internal fixation is of secondary importance. PMID- 10790672 TI - Paget's Disease of Bone: Pathophysiology, Diagnosis, and Management. AB - Paget's disease of bone is a common geriatric disorder of skeletal remodeling, which may have a viral etiology. Safe and effective treatments are now available for associated complications of symptomatic involvement. The orthopaedic surgeon should have a fundamental understanding of the complications of Paget's disease and should be familiar with the indications for treatment, as well as available medical and surgical therapies. PMID- 10790673 TI - Thoracolumbar Spine Trauma: I. Evaluation and Classification. AB - A timely and thorough evaluation of thoracolumbar injuries and rational treatment based on a complete understanding of the mechanism of bone, soft-tissue, and nerve injury is essential for maximizing the patient's neurologic and functional recovery and minimizing associated complications, the time to recovery, and the problems of long-term pain and deformity. The initial evaluation includes both clinical and radiologic assessment. Clinical evaluation includes the general trauma examination as well as a detailed spinal and neurologic examination to determine the level (or levels) of spinal injury. Radiologic evaluation includes both plain radiography and the appropriate use of advanced imaging modalities. A review of the evolution of thoracolumbar injury classifications is presented. PMID- 10790674 TI - Thoracolumbar Spine Trauma: II. Principles of Management. AB - The care of patients with thoracolumbar spine trauma with or without neurologic deficits has evolved dramatically over the past 20 years with the emergence of tertiary-care spinal injury centers and the development of more effective spinal instrumentation and anesthesia techniques. Despite these advances, the majority of patients with thoracolumbar injuries are still treated nonoperatively with cast or brace immobilization and early ambulation. More aggressive treatment is guided by the use of classification systems that detail the mechanism of injury, the degree of compromise of spinal structures, and the potential for late mechanical instability or neural injury. The goal of treatment remains attainment of spinal stability with protection or improvement of the patient's neurologic status, allowing rapid and maximal functional recovery. PMID- 10790675 TI - Clavicular Nonunion and Malunion: Evaluation and Surgical Management. AB - Nonunions and malunions of the clavicle are uncommon but can be disabling. Pain, limitation of shoulder mobility, or local compression of the brachial plexus can produce profound functional impairment. Nonunions usually are associated with more severe fractures, open injuries, or failures of operative treatment. Reconstructive procedures are focused on gaining union and restoring clavicular anatomy-most often achieved with plates and screws and autogenous bone graft. Salvage procedures include excision of a bony prominence, partial or total clavicular resection, and resection of the first rib. While most patients with a malunited clavicular fracture are asymptomatic, osteotomy and correction of the deformity should be considered when there is associated functional or neurovascular impairment. PMID- 10790676 TI - Legg-Calve-Perthes Disease. AB - Legg-Calve,-Perthes disease is a self-limited disease of the femoral head that presents in the first decade. The pathogenesis is thought to involve bone necrosis, collapse, and repair. The presenting complaint is often a painless limp or hip pain, with decreased abduction and internal rotation of the hip. Factors that are believed to correlate with a poor prognosis are onset of symptoms after age 8 years, lateral head subluxation, involvement of over 50% of the femoral head with collapse of the lateral pillar, and the combination of an aspherical femoral head and an incongruent joint. The current cornerstones of treatment are maintenance of hip motion, relief of symptoms, and containment. Containment may be achieved by bracing or surgical means. The literature remains inconclusive on the indications for and effects of treatment. A long-term study has suggested that disabling arthritis of the hip develops in the sixth decade of life in 50% of untreated patients. PMID- 10790677 TI - Evaluating multiple treatment courses in clinical trials. AB - In oncology, a patient's treatment often involves multiple courses of chemotherapy. The most common medical practice in choosing treatments for successive courses is to repeat a treatment that is successful in a given course and otherwise switch to a different treatment. Patient outcome thus consists of a sequence of dependent response variables and corresponding treatments. Despite the widespread use of such adaptive 'play-the-winner-and-drop-the-loser' algorithms in medical settings involving multiple treatment courses, most statistical methods for treatment evaluation characterize early patient outcome as a single response to a single treatment, resulting in a substantial loss of information. In this paper, we provide a statistical framework for multi-course clinical trials involving some variant of the play-the-winner-and-drop-the-loser strategy. The aim is to design and conduct the trial to more closely reflect actual clinical practice, and thus increase the amount of information per patient. The proposed design is similar to a multi-stage cross-over trial, with the essential difference that here all treatments after the first course are assigned adaptively. We illustrate the method by application to a randomized phase II trial for androgen independent prostate cancer. We consider the goals of selecting one best treatment, or selecting a best ordered pair of treatments with the second given if the first fails to achieve a patient success. A simulation study is reported, and extensions to trials involving toxicity or regimen-related death are discussed. PMID- 10790678 TI - On statistical analysis for placebo-challenging designs in clinical trials. AB - In clinical trials, appropriate designs are often chosen to address scientific/medical questions of particular interest to the investigator. For a chosen statistical design, however, standard statistical procedures may not be applicable owing to the nature of the design. In this paper we examine statistical methods for analysis of data collected from a placebo-challenging design which is often considered for assessment of the efficacy of drug products for indication of erectile dysfunction. An example concerning a clinical trial conducted with 120 male patients with erectile dysfunction is used to illustrate the derived statistical methods. Some recommendations to the randomization procedure for the study design of this kind are also made. PMID- 10790679 TI - Non-parametric covariance methods for incidence density analyses of time-to-event data from a randomized clinical trial and their complementary roles to proportional hazards regression. AB - The principal response criteria for many clinical trials involve time-to-event variables. Usual methods of analysis for this type of response criterion include product-limit estimators of cumulative survival for the treatment groups, (stratified) logrank tests to compare treatments, and proportional hazards regression models with treatment and relevant covariates. When adjustment for covariates is of some importance, the relative roles of these methods may be of some concern, particularly for confirmatory clinical trials which must provide convincing findings to regulatory agencies. Unadjusted methods may have lower power, but there are issues regarding adjustment for covariates that may be controversial. These issues include applicability of proportional hazards assumptions, whether the correct model has been specified, and whether there is parallelism between treatments for relationships with covariates. One way to address these issues is to use non-parametric analysis of covariance strategies with extensions to log incidence density estimation. The principal basis for this method is no association between covariates and treatment groups as provided by randomized assignment of patients to groups. The background theory and strategies for computation are described for this method. Aspects of its application are illustrated for a clinical trial with two treatment groups and 722 patients. The objective of analysis for this clinical trial is evaluation of treatment effects with and without adjustment for 22 a priori covariates and a stratification for three geographical regions. PMID- 10790680 TI - Prognostic modelling with logistic regression analysis: a comparison of selection and estimation methods in small data sets. AB - Logistic regression analysis may well be used to develop a prognostic model for a dichotomous outcome. Especially when limited data are available, it is difficult to determine an appropriate selection of covariables for inclusion in such models. Also, predictions may be improved by applying some sort of shrinkage in the estimation of regression coefficients. In this study we compare the performance of several selection and shrinkage methods in small data sets of patients with acute myocardial infarction, where we aim to predict 30-day mortality. Selection methods included backward stepwise selection with significance levels alpha of 0.01, 0.05, 0. 157 (the AIC criterion) or 0.50, and the use of qualitative external information on the sign of regression coefficients in the model. Estimation methods included standard maximum likelihood, the use of a linear shrinkage factor, penalized maximum likelihood, the Lasso, or quantitative external information on univariable regression coefficients. We found that stepwise selection with a low alpha (for example, 0.05) led to a relatively poor model performance, when evaluated on independent data. Substantially better performance was obtained with full models with a limited number of important predictors, where regression coefficients were reduced with any of the shrinkage methods. Incorporation of external information for selection and estimation improved the stability and quality of the prognostic models. We therefore recommend shrinkage methods in full models including prespecified predictors and incorporation of external information, when prognostic models are constructed in small data sets. PMID- 10790681 TI - Are standardized mortality ratios valid for public health data analysis? AB - Standardized mortality ratios (SMRs) have been criticized as lacking validity, and it has been recommended to use standardized rate ratios (SRRs) instead. A review of the epidemiology literature and standard epidemiology textbooks showed disagreement concerning the validity of SMRs and a lack of data to support claims concerning their validity. Therefore, we sought to determine the validity of SMRs in public health data analysis. Simulations were carried out using widely disparate study population age distributions and disease rates encountered in public health data analysis. We compared SMRs and SRRs as absolute measures of increased mortality in a population, and for ranking mortality in different populations. The simulations showed that SMRs changed by 6 per cent to 8 per cent when the age distribution was changed from that of a 'young' age distribution to that of an 'old' age distribution. In comparison, SRRs changed by 4 per cent to 5 per cent when the age-adjustment standard was changed from the 1940 U.S. Census population to the 1990 U.S. Census population. County rankings by SRR were somewhat more similar among themselves than when compared with rankings by SMR, but the differences were not large. Based on our findings, SMRs are of similar usefulness to SRRs in public health data analysis, will lead to similar conclusions, and may be used to compare different geographic areas. PMID- 10790682 TI - A comparison of several procedures to estimate the confidence interval for attributable risk in case-control studies. AB - The estimation of a confidence interval for attributable risk from the logistic model based on data from case-control studies is a problem for which an accepted solution is lacking. Two methods, one based on the delta method and one bootstrap on the population base, have been described but their accuracy has not been compared. We present two other methods, one based on a jack-knife approach and the other using a bootstrap on two samples (cases and controls). The four methods are compared in a simulation study. The four methods are also applied to a case control study on risk factors for preterm delivery; the confidence intervals are obtained assuming normality and by logarithmic transformation. When attributable risk is not smooth (for example, when exposure prevalence is low) both the jack knife and the delta method tend to fail. If attributable risk is close to zero or one, normality cannot be assumed and log-transformed confidence intervals must be used. Finally, the extension to matched studies is analysed using a case-control study on risk factors of cutaneous malignant melanoma. In this situation, the population-based bootstrap is not available. PMID- 10790683 TI - Tests for gaussian repeated measures with missing data in small samples. AB - For small samples of Gaussian repeated measures with missing data, Barton and Cramer recommended using the EM algorithm for estimation and reducing the degrees of freedom for an analogue of Rao's F approximation to Wilks' test. Computer simulations led to the conclusion that the modified test was slightly conservative for total sample size of N=40. Here we consider additional methods and smaller sample sizes, Nin?12,24?. We describe analogues of the Pillai Bartlett trace, Hotelling-Lawley trace and Geisser-Greenhouse corrected univariate tests which allow for missing data. Eleven sample size adjustments were examined which replace N by some function of the numbers of non-missing pairs of responses in computing error degrees of freedom. Overall, simulation results allowed concluding that an adjusted test can always control test size at or below the nominal rate, even with as few as 12 observations and up to 10 per cent missing data. The choice of method varies with the test statistic. Replacing N by the mean number of non-missing responses per variable works best for the Geisser-Greenhouse test. The Pillai-Bartlett test requires the stronger adjustment of replacing N by the harmonic mean number of non-missing pairs of responses. For Wilks' and Hotelling-Lawley, an even more aggressive adjustment based on the minimum number of non-missing pairs must be used. PMID- 10790684 TI - Strategies for comparing treatments on a binary response with multi-centre data. AB - This paper surveys methods for comparing treatments on a binary response when observations occur for several strata. A common application is multi-centre clinical trials, in which the strata refer to a sample of centres or sites of some type. Questions of interest include how one should summarize the difference between the treatments, how one should make inferential comparisons, how one should investigate whether treatment-by-centre interaction exists, how one should describe effects when interaction exists, whether one should treat centres and centre-specific treatment effects as fixed or random, and whether centres that have either 0 successes or 0 failures should contribute to the analysis. This article discusses these matters in the context of various strategies for analysing such data, in particular focusing on special problems presented by sparse data. PMID- 10790685 TI - Dynamic in vivo (31)P nuclear magnetic resonance study of Saccharomyces cerevisiae in glucose-limited chemostat culture during the aerobic-anaerobic shift. AB - The purpose of this work was to analyse in vivo the influence of sudden oxygen depletion on Saccharomyces cerevisiae, grown in glucose-limited chemostat culture, using a recently developed cyclone reactor coupled with (31)P NMR spectroscopy. Before, during and after the transition, intracellular and extracellular phosphorylated metabolites as well as the pHs in the different cellular compartments were monitored with a time resolution of 2.5 min. The employed integrated NMR bioreactor system allowed the defined glucose-limited continuous cultivation of yeast at a density of 75 g DW/l and a p(O(2)) of 30% air saturation. A purely oxidative metabolism was maintained at all times. In vivo (31)P NMR spectra obtained were of excellent quality and even allowed the detection of phosphoenolpyruvate (PEP). During the switch from aerobic to anaerobic conditions, a rapid, significant decrease of intracellular ATP and PEP levels was observed and the cytoplasmic pH decreased from 7.5 to 6.8. This change, which was accompanied by a transient influx of extracellular inorganic phosphate (P(i)), appeared to correlate linearly with the decrease of the ATP concentration, suggesting that the cause of the partial collapse of the plasma membrane pH gradient was a reduced availability of ATP. The complete phosphorous balance established from our measurement data showed that polyphosphate was not the source of the increased intracellular P(i). The derived intracellular P(i), ATP and ADP concentration data confirmed that the glycolytic flux at the level of glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase and enolase enzymes is mainly controlled by thermodynamic constraints. PMID- 10790686 TI - Biotransformation of monoterpene alcohols by Saccharomyces cerevisiae, Torulaspora delbrueckii and Kluyveromyces lactis. AB - Monoterpenoids are important flavour compounds produced by many plant species, including grapes (Vitis vinifera) and hops (Humulus lupulus). Biotransformation reactions involving monoterpenoids have been characterized in filamentous fungi, but few examples have been observed in yeasts. As monoterpenoids are in contact with yeasts during beer and wine production, biotransformation reactions may occur during the fermentation of these beverages. This paper describes the biotransformation of monoterpene alcohols, of significance in the alcoholic beverage industries, by three yeast species. All three species analysed had the ability to convert monoterpenoids. Saccharomyces cerevisiae and Kluyveromyces lactis reduced geraniol into citronellol, whilst all three yeasts produced linalool from both geraniol and nerol. Monocyclic alpha-terpineol was formed from both linalool and nerol, by all three yeasts. alpha-Terpineol was then converted into the diol cis-terpin hydrate. K. lactis and Torulaspora delbrueckii also had the ability to form geraniol from nerol. Finally, the stereospecificity of terpenoid formation was analysed. Both (+) and (-) enantiomers of linalool and alpha-terpineol were formed in roughly equal quantities, from either geraniol or nerol. PMID- 10790687 TI - A comprehensive web resource on RNA helicases from the baker's yeast Saccharomyces cerevisiae. AB - Members of the RNA helicase protein family are defined by several motifs that have been widely conserved during evolution. They are found in all organisms-from bacteria to humans-and many viruses. The minimum number of RNA helicases present within a eukaryotic cell can be predicted from the complete sequence of the Saccharomyces cerevisiae genome. Recent progress in the functional analysis of various family members has confirmed the significance of RNA helicases for most cellular RNA metabolic processes. We have assembled a web resource that focuses on RNA helicases from the budding yeast Saccharomyces cerevisiae. It includes descriptions of RNA helicases and their functions, links to sequence- and yeast specific databases, an extensive list of references, and links to non-yeast helicase web resources. PMID- 10790688 TI - Two-dimensional gel analysis of the proteome of lager brewing yeasts. AB - Modern lager brewing yeasts used in beer production are hybrid strains consisting of at least two different genomes. To obtain information on the identity of the parental strains that gave rise to industrial lager yeasts, we used two dimensional (2-D) gel electrophoresis and analysed the proteomes of different Saccharomyces species isolated from breweries. We found that the proteome of lager brewing yeasts and of the type strains of S. carlsbergensis, S. monacensis and S. pastorianus can be interpreted as the superimposition of two elementary patterns. One originates from proteins encoded by a S. cerevisiae-like genome. The other corresponds to a divergent Saccharomyces species whose best representative is a particular S. pastorianus strain, NRRL Y-1551. A map of industrial lager brewing yeasts has been established, with the individual origin of proteins and with identification of protein spots by comparison to known S. cerevisiae proteins. This 2-D map can be accessed on the Lager Brewing Yeast Protein Map server through the World Wide Web. This study provides the first example of the use of proteome analysis for investigating taxonomic relationships between divergent yeast species. PMID- 10790689 TI - Cyclic AMP regulates cell size of Schizosaccharomyces pombe through Cdc25 mitotic inducer. AB - Nutritional state modulates the cell size of the fission yeast Schizosaccharomyces pombe, such that cells grown in rich medium are larger in size than those in poor medium. This signal is transduced partly through the cyclic AMP-dependent protein kinase cascade. However, little is known about how cyclic AMP interacts with the central cell cycle machinery, Cdc2, the cyclin dependent kinase that induces mitosis. We show here that cyclic AMP regulates mitosis and cell size, in part, through regulation of protein stability of the Cdc2-activating phosphatase, Cdc25. However, our analysis demonstrates that cyclic AMP can negatively regulate mitosis independently of dephosphorylation of Cdc2 at Tyr(15). PMID- 10790690 TI - Molecular cloning of the CRM1 gene from Candida albicans. AB - In a screen for Candida albicans genes capable of supressing a ste20Delta mutation in Saccharomyces cerevisiae, a homologue of the exportin-encoding gene CRM1 was isolated. The CaCRM1 gene codes for a protein of 1079 amino acids with a predicted molecular weight of 124 029 and isoelectric point of 5.04. Crm1p from C. albicans displays significant amino acid sequence homology with Crm1p from Saccharomyces cerevisiae (65% identity, 74% similarity), Schizosaccharomyces pombe (55% identity, 66% similarity), Caenorhabditis elegans (45% identity, 57% similarity), and Homo sapiens (48% identity, 59% similarity). Interestingly, CaCRM1 encodes a threonine rather than a cysteine at position 533 in the conserved central region, suggesting that CaCrm1p is leptomycin B-insensitive, like S. cerevisiae Crm1p. CaCRM1 on a high copy vector can complement a thermosensitive allele of CRM1 (xpo1-1) in S. cerevisiae, showing that CaCrm1p and S. cerevisiae Crm1p are functionally conserved. Southern blot analysis suggests that CaCRM1 is present at a single locus within the C. albicans genome. The nucleotide sequence of the CaCRM1 gene has been deposited at GenBank under Accession No. AF178855. PMID- 10790691 TI - Identification by functional analysis of the gene encoding alpha-isopropylmalate synthase II (LEU9) in Saccharomyces cerevisiae. AB - The function of the open reading frame (ORF) YOR108w of Saccharomyces cerevisiae has been analysed. The deletion of this ORF from chromosome XV did not give an identifiable phenotype. A mutant in which both ORF YOR108w and LEU4 gene have been deleted proved to be leucine auxotrophic and alpha-isopropylmalate synthase (alpha-IPMS)-negative. This mutant recovered alpha-IPMS activity and a Leu(+) phenotype when transformed with a plasmid copy of YOR108w. These data and the sequence homology indicated that YOR108w is the structural gene for alpha-IPMS II, responsible for the residual alpha-IPMS activity found in a leu4Delta strain. The leu4Delta strain appeared to be very sensitive to the leucine analogue trifluoroleucine. In the absence of leucine, its growth was not much impaired in glucose but more on non-fermentable carbon sources. PMID- 10790692 TI - Systematic disruption of 456 ORFs in the yeast Saccharomyces cerevisiae. AB - In the framework of the European Network for Functional Analysis (EUROFAN), five packages of 96 ORFs from chromosomes III, IV, VII, XIII, XIV and XV were subjected to systematic deletions in an isogenic derivative of strain S288c. Deletions were constructed in diploid and haploid strains. Two questionable ORFs overlapping with larger ORFs and seven TY ORFs were discarded. A total of 456 heterozygous and 385 homozygous deletant diploids were obtained. Sixty-nine deletions, 25 of which had never been published before, were lethal in haploid strains and 30 caused slow cellular growth. PMID- 10790693 TI - A counterselection for the tryptophan pathway in yeast: 5-fluoroanthranilic acid resistance. AB - The ability to counterselect, as well as to select for, a genetic marker has numerous applications in microbial genetics. Described here is the use of 5 fluoroanthranilic acid for the counterselection of TRP1, a commonly used genetic marker in the yeast Saccharomyces cerevisiae. Counterselection using 5 fluoroanthranilic acid involves antimetabolism by the enzymes of the tryptophan biosynthetic pathway, such that trp1, trp3, trp4 or trp5 strains, which lack enzymes required for the conversion of anthranilic acid to tryptophan, are resistant to 5-fluoroanthranilic acid. Commonly used genetic procedures, such as selection for loss of a chromosomally integrated plasmid, and a replica-plating method to rapidly assess genetic linkage in self-replicating shuttle vectors, can now be carried out using the TRP1 marker gene. In addition, novel tryptophan auxotrophs can be selected using 5-fluoroanthranilic acid. PMID- 10790695 TI - Current awareness on yeast. PMID- 10790694 TI - The products of YCF1 and YLL015w (BPT1) cooperate for the ATP-dependent vacuolar transport of unconjugated bilirubin in Saccharomyces cerevisiae. AB - Since bilirubin-like pigments are present in the environment as degradation products of heme-containing proteins, yeast could have developed a detoxifying system to transport these compounds into their vacuoles. Vacuoles from Saccharomyces cerevisiae showed an ATP-dependent, saturative transport of unconjugated bilirubin (UCB) that was reduced by 60% and 40% in YCF1 and YLL015w deleted cells, respectively; the double deletant showed no UCB uptake. Conversely, the transport of bile acids (taurocholate) was comparable in wild and deleted stains. These data identify YCF1 and YLL015w, named BPT1 (Bile Pigment Transporter), as the genes responsible for ATP-dependent UCB transport in yeast. Since YCF1 and YLL015w are rather homologous with multidrug resistant proteins (MRPs), they also suggest the involvement of this class of transporters in the ATP-dependent transport of unconjugated bilirubin. PMID- 10790697 TI - Health economics: an evolving paradigm. PMID- 10790698 TI - Centre-specific or average unit costs in multi-centre studies? Some theory and simulation. AB - Costing issues are increasingly being addressed in multi-centre studies. In this paper, two methods for collecting costing information are compared within a simulated clinical trial setting. One method estimates average treatment costs by applying unit costs averaged across treatment centres to centre-specific volumes of resource use. The second uses centre-specific information for both the unit costs and the resource volumes, and then averages across centres. Using a pre specified production relation between the different volumes of resource use, and simulating changes in unit costs, it is shown that these two methods result in statistically different estimates of average treatment costs. This finding holds, regardless of the degree of substitutability between the resource volumes, except when considerable uncertainty surrounds treatment centre responses to relative changes in unit costs. The findings suggest that a more cautious approach should be adopted in the collection, calculation and interpretation of treatment costs in multi-centre studies. PMID- 10790699 TI - The effect of capitation on GPs' referral decisions. AB - In the Norwegian capitation trial, the payment system for general practitioners (GPs) has been changed; a practice allowance component has been replaced by a capitation component and the fee-per-item component constitutes a smaller part of a GPs practice income than previously. From the theoretical modelling of GPs' referral decisions, we predict that the replacement of the practice allowance by a capitation component will increase the rate of referrals to specialists. This hypothesis is supported by an exploratory empirical study with data from a sample of GPs participating in the experiment. PMID- 10790700 TI - Can cost shifting continue in a price competitive environment? AB - Both Medicare and Medicaid are reducing payments to hospitals, and there is widespread concern that hospitals may respond by increasing prices to privately insured patients. Theoretical models of hospital behaviour have ambiguous predictions as to whether, and under what circumstances, hospitals will shift costs to private payers. This paper extends previous theoretical models and then tests empirically using data from California for the 1983-1991 period, a time of increasingly intense price competition. Hospitals did increase their prices to private payers in response to reductions in Medicare rates; they had far smaller and generally insignificant responses to changes in Medicaid reimbursement. Hospital ownership and the competitiveness of the hospital market both affected this behaviour, but there was no significant change over time. The results suggest the need to broaden our models of hospital behaviour to 'embed' them in their local markets. PMID- 10790701 TI - Power and sample assessments for tests of hypotheses on cost-effectiveness ratios. AB - We address the issue of statistical power and sample size for cost-effectiveness studies. Tests of hypotheses on the cost-effectiveness ratio (CER) are constructed from the net cost and incremental effectiveness measures. When the difference in effectiveness is known, we derive formulae for statistical power and sample size assessments for one- and two-sided tests of hypotheses of the CER. We also construct a test of the joint hypothesis of cost-effectiveness and effectiveness and derive an expression connecting power and sample size. Our methods account for the correlation between cost and effectiveness and lead to smaller sample size requirements than comparative methods that ignore the correlation. The implications of our formulae for cost-effectiveness studies are illustrated through numerical examples. When compared with trials designed to demonstrate effectiveness alone, our results indicate that a trial appropriately powered to demonstrate cost-effectiveness might require sample sizes many times greater. PMID- 10790703 TI - Uncertainty aversion: a reply to the paper by Andersson and Lyttkens. PMID- 10790704 TI - Uncertainty aversion-a reply to oliver PMID- 10790702 TI - Development of WHO guidelines on generalized cost-effectiveness analysis. AB - The growing use of cost-effectiveness analysis (CEA) to evaluate specific interventions is dominated by studies of prospective new interventions compared with current practice. This type of analysis does not explicitly take a sectoral perspective in which the costs and effectiveness of all possible interventions are compared, in order to select the mix that maximizes health for a given set of resource constraints. WHO guidelines on generalized CEA propose the application of CEA to a wide range of interventions to provide general information on the relative costs and health benefits of different interventions in the absence of various highly local decision constraints. This general approach will contribute to judgements on whether interventions are highly cost-effective, highly cost ineffective, or something in between. Generalized CEAs require the evaluation of a set of interventions with respect to the counterfactual of the null set of the related interventions, i.e. the natural history of disease. Such general perceptions of relative cost-effectiveness, which do not pertain to any specific decision-maker, can be a useful reference point for evaluating the directions for enhancing allocative efficiency in a variety of settings. The proposed framework allows the identification of current allocative inefficiencies as well as opportunities presented by new interventions. PMID- 10790705 TI - Uncertainty aversion: a discussion of critical issues in health economics. PMID- 10790706 TI - Announcement: health economics letters PMID- 10790707 TI - The veil of experience: do consumers prefer what they know best? AB - There is growing interest from health policy makers in eliciting consumer preferences for health care services. This is particularly the case when assessing the likely impact of innovations. Some people may be wary of innovations because they prefer the service they have previously experienced. Consumer preferences for an existing and a hypothetical new bowel cancer testing programme were measured using a discrete choice experiment questionnaire. The results showed that consumers had a statistically significant preference for the existing service (status quo) when all other factors remained constant. It suggested that consumers make decisions under a 'veil of experience'. Possible explanations for this result include the endowment effect, status quo bias and loss aversion. Future evaluations of health service innovation should be aware of this tendency to favour the status quo. PMID- 10790708 TI - A. V. Hernandez, 'Duplicate coverage and demand for health care. The case of Catalonia'. Health economics 8(7) 1999, 579-598 AB - The original article to which this Erratum refers was published in Health Economics 8(7) 1999, 579-598. PMID- 10790709 TI - Bridging the policy-practice gap. PMID- 10790710 TI - Balancing stakeholder needs: a review of ENB 100 and 415 courses. English National Board for Nursing, Midwifery and Health Visiting. AB - This paper reports the findings of a documentary analysis and literature review of general and paediatric intensive care unit (ICU) courses (ENB 100 and ENB 415). The findings are part of a larger review of critical care courses commissioned by the English National Board for Nursing, Midwifery and Health Visiting (ENB), also incorporating operating department, coronary care and accident and emergency courses. It was important to set the curriculum review in the context of intensive care practice and education, hence the study also comprised interviews with lecturers and ICU managers. The study findings reveal diversity in major aspects of the critical care courses, including the academic level of the programmes and credits they attracted; the assessment strategies for theory and practice, the extent of shared learning and the amount of student effort. Many factors influenced this diversity including contrary opinion among stakeholders about the purpose of the course: to prime the students for working in the specialty; or to consolidate previous experience (in some cases up to 15 years). Course structure and content have changed in response to local university requirements and directives from the statutory bodies, as well as in response to the higher level of academic credit awarded for pre-registration programmes (qualification inflation). The perceived shift in course content as well as the diversity across programmes had led a group of ICU managers to define their own list of competencies (Crunden 1998). However, the majority of the managers interviewed for this study (63% of General ICU managers (n = 19) and 83% (n = 6) of Paediatric ICU managers) were generally satisfied with the competencies and skills of the nurses who had undertaken the ENB course. The authors conclude from the diverse nature of the courses that there is little national comparability in the courses although this finding might be an artefact of documentary analysis. The extent to which this (apparent) diversity results in different levels of competence in practice requires further exploration. PMID- 10790711 TI - Developing specialty knowledge: the case of Australian critical care nursing. AB - The generation of new knowledge is important to the evolution of the nursing profession and its specialties and is a fundamental requirement for the development of the discipline. This study documents the growth of empirical knowledge in Australian critical care nursing by examining the research published in one critical care specialty journal and a second generalized nursing research journal between 1994 and 1998. The proportion of research published in the specialty journal increased from 25 to 69%, with almost one-half focusing on patient therapies and treatments. The proportion of research in the general journal was consistently high, approximately 80 to 90%, with one to two critical care research articles per year. Nurses were first authors on the vast majority of these research articles. Approximately half of the critical care research focused on patient care. The results support the claim that Australian critical care nurses are conducting research to promote best practice in patient care and are thus contributing to the development of the discipline of nursing. PMID- 10790712 TI - Quality of life and the high-dependency unit. AB - This study was designed to identify and measure the patients' perspective of the concept quality of life within the context of a high-dependency unit (HDU). Data were collected in two phases. In phase one, 55 patients were interviewed, which resulted in the concept clarification of quality of life as: physical, social, psychological and family/friends. In phase two, 51 patients undertook quality-of life assessment using validated instruments the Quality of Life Index (Ferrans & Powers 1985) and the Global Quality of Life Scale (Hyland & Sodergren 1997). Post HDU patients demonstrated improvements upon pre-admission scores in both instruments (the social domain in the quality of life index being the exception), although this only reached statistical significance P<0.05 in the overall index score and within the domains of health and family. When exploring variables of age and severity of illness (Apache 2 score, Knaus et al. 1980) it was the people who were older and physiologically compromised to an increased extent (P<0.05) who demonstrated higher levels of satisfaction with their resulting quality of life. This study has been successful in providing patients with an opportunity to participate more actively in service evaluation and has identified the need for future evaluation of HDUs to move beyond physiological measures, to incorporate the impact that illness leading to admission to an HDU has upon the patients' ability to function and their resulting quality of life. PMID- 10790713 TI - Optimizing oxygen delivery: haemodynamic workshop. Part 2. PMID- 10790714 TI - Meeting the needs of the family: the role of the specialist nurse in the management of brain death. AB - Clinical nurse specialist roles have existed in the USA for many years, yet here in the UK such roles are a relatively new concept for nurses and the profession of nursing. In the UK clinical nurse specialists have worked primarily in outpatient settings, however, the role is rapidly developing in primary care and specialized care settings. One specific dimension of the clinical nurse specialist role in critical care relates to the needs of the family when brain stem death is suspected. This concept of death--brain death--is very difficult for most people to comprehend as it challenges all our previous beliefs concerning death and dying. During this traumatic time nurses and other health care professionals face the daunting task of supporting the family through this enforced emotional crisis as they attempt to come to terms with the diagnosis of clinical brain death. To enable families to understand brain death and the implications and outcomes of such a diagnosis it is vital they receive information, support and guidance from experienced health care personnel. The following account is a discussion of the key role of the specialist nurse in relation to meeting the needs of the families of brain-stem dead patients. This review seeks to highlight the clear dimensions of the clinical nurse specialist role within a critical care setting. It relates specifically to meeting the needs of families who have experienced clinical brain death in an immediate family member. It suggests that the infinite value of the role must be acknowledged and utilized to ensure the provision of holistic care for patients and families at this traumatic time. PMID- 10790717 TI - Chest X-ray quiz. PMID- 10790715 TI - Our journey with clinical supervision in an intensive care unit. AB - In 1991, the nursing profession began to pursue the idea of reflective practice with vigour, despite little evidence that reflective practice improved nursing care (Burnard 1991; Hunt 1993). A small qualitative study on the experiences of nurses writing reflective journals was undertaken in the intensive care unit at Newham General Hospital. (A Price 1995). This highlighted the need for clear initial guidance and support when introducing reflective practice. It also demonstrated a lack of knowledge about reflection within the unit. During this time, a group of nurses who were interested in developing clinical supervision as a method of support for staff was formed. Kohner's (1994) definition of clinical supervision, adopted by the group after review of literature, makes it clear that this involves reflection on practice. Johns' (1993) Model of Structured Reflection was amended and used to promote in-depth analysis of situations, and journal writing was encouraged within the group. A continuing process of group clinical supervision was started, initially with the help of a facilitator experienced in it. On the basis of this experience, a strategy for implementation of clinical supervision in the intensive care unit was developed and put into action by the group members. Consideration of the progress and problems experienced leads to the conclusion that continuing staff motivation and commitment, and adequate time are essential for implementation of clinical supervision. PMID- 10790718 TI - Temozolomide (Temodal) for treatment of primary brain tumours. AB - Although the outlook for patients with malignant gliomas continues to be very poor, progress in neurosurgery and radiotherapy, supported by new development in chemotherapy, offer some hope for the future. Temozolomide has recently been introduced by Schering-Plough Ltd (Welwyn Garden City, UK) as a new treatment which merits further investigation in this situation. PMID- 10790719 TI - A preliminary understanding of mania: roles for melatonin, vasotocin and rapid eye-movement sleep. AB - Speculation about mania links melatonin, vasotocin and rapid-eye-movement (REM) sleep. Normal REM sleep can have the loss of reality testing and this feature intruding into the wake period could be the cause of the distorted reality present in manic delusions. REM sleep has a role in memory formation. Abnormal levels and/or rhythms of melatonin are thought to be involved in mania. Decreased melatonin production with malfunction of REM sleep is proposed to be a cause of sudden infant death syndrome and Alzheimer's disease, because the loss of the antioxidant function of melatonin allows brain injury to occur. PMID- 10790720 TI - Mechanics of nucleocytoplasmic exchanges mediated by vertebrates' nuclear pore complex. AB - The paper proposes a mechanism for regulating nucleo-cytoplasmic exchanges of macromolecules via NPC in vertebrates. Bending of NE toward the transport vector significantly contributes to the dynamics. PMID- 10790721 TI - Activation of human neutrophils by electronically transmitted phorbol-myristate acetate. AB - We report the transfer of the activity of 4-phorbol-12-beta-myristate-13-acetate (PMA) by electronic means. Neutrophils were placed at 37 degrees C on one coil attached to an oscillator, while PMA was placed on another coil at room temperature. The oscillator was then turned on for 15 min, after which cells were usually further incubated for up to 45 min at 37 degrees C before measurement of reactive oxygen metabolites (ROMs) production. In 20 blind experiments, PMA thus 'transmitted' induced ROM production. ROM were not induced when: (1) PMA vehicle or 4-alpha-phorbol 12,13-didecanoate (an inactive PMA analogue) were transmitted; (2) the oscillator was switched off; (3) superoxide dismutase or protein kinase C inhibitors were added to cells before transmission. These results suggest that PMA molecules emit signals that can be transferred to neutrophils by artificial physical means in a manner that seems specific to the source molecules. PMID- 10790722 TI - Electrical and chemical synaptic transmission as an interacting system. AB - It is proposed that presynaptic potassium efflux triggered by the nerve impulse may generate either excitatory or inhibitory responses depending on the neurotransmitter which more or less steadily impregnates the postsynaptic membrane. The jelly intersynaptic matrix may potentiate the efficiency of inoic intersynaptic signals. The synaptic vesicles are proposed to shuttle mitochondrial ATP towards the presynaptic membrane, thereby supplying the energy necessary to restore the membrane polarity after synaptic transmission. Plain structural data and currently accepted functional antecedents appear to justify the proposal. PMID- 10790723 TI - Repositioning of entangled chromatin during separation of sister chromatids by eversion. AB - We investigate how chromatin entanglements may subvert the topology of DNA segments (and of genes therein) whenever sister chromatids separate by eversion. Such subversions offer interesting ideas on how identical genes may be differently or identically expressed after segregation of sister chromatids. PMID- 10790724 TI - Theoretical aspects of the neurobiological integration of memory. AB - Whenever a peripheral structure like the visual system captures information, the input signal reverberates in circuits of neurons, which send it thereafter towards: (a) the motor system, triggering a specific response, evoked by a short term memory mechanism; and (b) the hippocampus, to produce long-term potentiation or depression. Two different processes regulate short-term memory: (1) Homosynaptic depression that inhibits neurotransmitter release by means of a decrease in Ca++ inflow, and an increase in calmodulin affinity for synaptic vesicles; and (2) Heterosynaptic facilitation that triggers neurotransmitter release, whenever serotonin activates a proteinkinase A. Besides carrying out a brief review on the matter, we support two different physiological explanations with regard to: (a) ion exchange process and the interstitial pH during habituation; and (b) the possibility of a sensitive presynaptic neuron interaction within the habituated reverberant circuit, to trigger dishabituation. We also propose the term 'time-mediated stimulatory action dependent' to name those serotonin receptors that may lead to a rapid or a delayed postsynaptic onset responses. PMID- 10790725 TI - The symptoms of chronic fatigue syndrome are related to abnormal ion channel function. AB - The pathogenesis of chronic fatigue syndrome (CFS) is unknown but one of the most characteristic features of the illness is fluctuation in symptoms which can be induced by physical and/or mental stress. Other conditions in which fluctuating fatigue occurs are caused by abnormal ion channels in the cell membrane. These include genetically determined channelopathies, e.g. hypokalemic periodic paralysis, episodic ataxia type 2 and acquired conditions such as neuromyotonia, myasthenic syndromes, multiple sclerosis and inflammatory demyelinating polyneuropathies. Our hypothesis is that abnormal ion channel function underlies the symptoms of CFS and this is supported also by the finding of abnormal cardiac thallium201 SPECT scans in CFS, similar to that found in syndrome X, another disorder of ion channels. CFS and syndrome X can have identical clinical symptoms. CFS may begin after exposure to specific toxins which are known to produce abnormal sodium ion channels. Finally, in CFS, increased resting energy expenditure (REE) occurs, a state influenced by transmembrane ion transport. The hypothesis that ion channels are abnormal in CFS may help to explain the fluctuating fatigue and other symptoms. PMID- 10790726 TI - Regulation of herpesvirus replication by subcellular compartmentalization. AB - The transcriptional regulation of herpesvirus gene expression has been well documented. A second model is proposed that is superimposed on regulation at the transcriptional level. The regulation is post-translational in nature. Three examples of the model are found in viral DNA replication, capsid assembly, and the cleavage and packaging of DNA into capsids. For each example, at least one viral protein depends upon an interaction with a second viral protein for transport into the nucleus. A model is proposed whereby these protein-protein interactions control the efficiency of these processes by the formation of the appropriate protein complexes in the cytoplasm. The model predicts that these interactions impose a necessary control and that mechanisms to bypass this control would deleteriously affect virus replication. It is probable that level of regulation extends for each of these processes among other herpesviruses. PMID- 10790727 TI - Prion diseases: a typical Kuhnian abnormality in a molecular paradigm. AB - As a new class of pathogens with unusual properties, prions have been implied in several spongiform encephalopathies mainly affecting farm animals (scrapie, mad cow disease) and humans (kuru, Creutzfeldt-Jacob disease, fatal familial insomnia) (1). The term prions underlines the feature that the agents would merely consist of a protein (2), perhaps a pathological counterpart of a cellular constituent with poorly defined functions (3-5). In reviewing relevant literature, in order to test the validity of the 'protein-only hypothesis' for coherence and consistency with a molecular paradigm, we have met evident discrepancies, making prion diseases a typical abnormality in a Kuhnian sense. This paper summarizes and analyses the main features of the protein-only model. PMID- 10790728 TI - Bronchial asthma: information on phytotherapy with essential fatty acids. Interactions between essential fatty acids and steroid hormones. AB - The pharmacological treatment of bronchial asthma is based on the inhalation of bronchodilative beta2 agonists and steroid hormones. In view of the therapeutic successes obtained with evening primrose oil in the treatment of neurodermatitis in children, the question arises whether this phytotherapeutic substitution therapy could at least partly replace symptomatic pharmacotherapy in bronchial asthma. It is shown that this will only be successful if one dispenses with the anti-inflammatory effect of the treatment with steroid hormones. PMID- 10790729 TI - 5-Lipoxygenase gene polymorphism and onset of Alzheimer's disease. AB - Recently, inflammatory pathways have been recognized as possible pathophysiological mechanisms of aging-associated neurodegenerations, and slowing of the progression of Alzheimer's disease can be achieved with anti-inflammatory drugs. The formation of endogenous inflammatory lipid mediators, leukotrienes, is initiated by 5-lipoxygenase (5-LOX), which is also expressed in neurons. We recently reported that aging is associated with a significant increase in neuronal 5-LOX gene expression and with increased, 5-LOX inhibitor-sensitive, vulnerability of neurons to degeneration. On the basis of these findings we have proposed that the 5-LOX pathway may influence the progression of aging-associated diseases, such as Alzheimer's. In humans, mutations of the promoter of the 5-LOX gene occur in a normal population with a frequency of about 25%. These mutations result in a decreased expression of the 5-LOX gene. Thus, it is hypothesized here that the onset of Alzheimer's disease will be delayed in subjects who display a mutation in the 5-LOX gene promoter, and that consequently the frequency of occurrence of the three known 5-LOX mutated alleles will be greater in subjects with onset of Alzheimer's at a very old age than in subjects with an earlier onset. PMID- 10790731 TI - A unifying hypothesis for the renin-angiotensin system and hematopoiesis: sticking the pieces together with the JAK-STAT pathway. AB - JAK-STAT pathway is a recently encountered intracellular signal transduction system. The pathway is utilized by numerous cytokines, growth factors, and hormones for gene expression and a variety of biological activities. Hematopoiesis is regulated by many cytokines and growth factors that support the proliferation and differentiation of progenitor cells in the bone marrow. JAK STAT pathway arises as the most common signalling cascade of a wide range of cytokines and/or growth factors in propagation of physiological and pathological/neoplastic hematopoiesis. On the other side, renin-angiotensin system (RAS) includes not only the classic circulating endocrine system controlling blood pressure and electrolyte homeostasis, but also tissue-specific RASs with autocrine and/or paracrine functions. Preliminary data suggest the involvement of the RAS components in normal and pathologic hematopoiesis, although the precise mechanism of action has not been elucidated yet. We have hypothesized, in this report, that JAK-STAT pathway serves as a point of crosstalk between the components of the locally present RAS in the bone marrow and hematopoiesis. Demonstration of a local RAS in the bone marrow with clarification of the postreceptor signalling events may play a consequential role not only for further clarification of normal hematopoiesis but also novel therapeutic approaches in pathologic/neoplastic conditions. PMID- 10790730 TI - For ischemic brain damage, is preclinical evidence of neuroprotection by presynaptic blockade of glutamate release enough? AB - Some drugs blocking glutamate release produce reduced brain injury in some animal models of cerebral ischemia whereas others lack a clear effect. Meta-analysis is a widely used technique in clinical and epidemiological studies. However, it has never been used in the analysis of preclinical studies. In order to estimate quantitatively the current state of the knowledge concerning the neuroprotective effect of drugs inhibiting glutamate release and to attempt to resolve the apparent controversy in relation to the neuroprotective properties of these drugs, a meta-analysis was performed. It identified a significant difference between drugs blocking glutamate release and controls. Therefore, we hypothesize that inhibition of presynaptic glutamate release could be a major goal in neuroprotection when ischemic brain damage is present and that meta-analysia could be a useful tool for preclinical studies, PMID- 10790732 TI - Sudden infant death syndrome: hypothalamic failure to sense elevated blood pyrogens. AB - Sudden infant death syndrome (SIDS) is frequently associated with a mild infection, the incidence peaking during the third month of life. We hypothesize that the neonatal immaturity of both the acute febrile response and hypothalamus promote neonatal protection from SIDS. Vagal afferents modify the febrile response. Vagotomized rodents displayed a loss of febrile responsiveness in a 'non-sensing' brain. The failure of a 'non- sensing' brain to react to elevated blood pyrogens leads to failure of the febrile response and to a shock-like state. SIDS infants may appear well yet, within hours of this observation, may be found dead. There is a mismatch between the acute febrile response and hypothalamic hypoactivation. The discrepancy increase wtih development. There is an elevated cytokine response in endothelial cells which induces nitric oxide (NO) production and retarded development of the hypothalamus. Cigarette smoke also induces NO production and retards hypothalamic development by augmented apoptosis. Zinc inhibits this effect in mouse thymocytes. Fetal haemoglobin (HbF) induces hypoxia which is a stimulator of the immune response, while vasodilator gases (carbon monoxide (CO), NO) reduce hypothalamic function. The hypothalamic failure to sense elevated blood pyrogens induces toxic shock--a feature of SIDS. PMID- 10790733 TI - Highly pleomorphic staphylococci as a cause of cancer. AB - An extensive historical literature exists suggesting that bacteria and other non virus microorganisms cause cancer. Much of this literature stresses the likely involvement of highly pleomorphic bacteria in carcinogenesis. Pleomorphic bacteria exhibit a variety of morphological types, some of which are identical to other bacteria. In particular, bacteria that can express more than one morphology, including that normally associated with common species of Staphylococcus, have frequently been isolated from cancers. Not surprisingly, this has led to considerable confusion and ridicule. The literature linking highly pleomorphic bacteria with carcinogenesis is presented here in an attempt to add weight to the view that bacteria, notably those expressing the morphology of common species of staphylococci, cause cancer. PMID- 10790734 TI - Theory of malignant cell transformation by superoxide fate coupled with cytoskeletal electron-transport and electron-transfer. AB - Signaling and tumor promoting functions have been experimentally assigned to the cytoskeleton, many of them linked to oxygen free radicals like superoxide. Superoxide and other reactive oxygen species (ROS) have been associated for many years with oncogenesis, and they are emerging as important signaling molecules connected to the classical signaling pathways, the cytoskeleton, the cell cycle control, and tumor initiation and promotion. Complex and multifunctional relationships between these entities are being discovered and attributed to specific protein-protein interactions. Theoretical analysis and experimental data indicate that small electronic currents may be carried by semiconduction electron transport along biopolymers. Therefore, it is proposed in this paper that the tumor-promoting effects mentioned above might be under control or modulation of these tiny electronic currents originated in relation to ROS and transported through the cytoskeletal actin microfilament network. PMID- 10790735 TI - Multiple sclerosis--in need of a critical reappraisal. AB - Multiple sclerosis (MS) is a disease that currently defies clinical and scientific definitions. Despite intensive clinical and basic research, very little is known about its possible cause(s) or pathogenesis, and the course and prognosis of MS practically remain unchanged. The aim of the present article is to outline some of the reasons for the constant failure to improve the therapy of MS. It also attempts to offer several guidelines which may enable a fresh and different approach to this devastating condition. PMID- 10790736 TI - Elevated, sustained peroxynitrite levels as the cause of chronic fatigue syndrome. AB - The etiology of chronic fatigue syndrome (CFS) has been both obscure and highly contentious, leading to substantial barriers to both clear diagnosis and effective treatment. I propose here a novel hypothesis of CFS in which either viral or bacterial infection induces one or more cytokines, IL-1beta IL-6, TNF alpha and IFN-gamma. These induce nitric oxide synthase (iNOS), leading to increased nitric oxide levels. Nitric oxide, in turn, reacts with superoxide radical to generate the potent oxidant peroxynitrite. Multiple amplification and positive feedback mechanisms are proposed by which once peroxynitrite levels are elevated, they tend to be sustained at a high level. This proposed mechanism may lower the HPA axis activity and be maintained by consequent lowered glucocorticoid levels. Similarities are discussed among CFS and autoimmune and other diseases previously shown to be associated with elevated peroxynitrite. Multiple pharmacological approaches to the treatment of CFS are suggested by this hypothesis. PMID- 10790737 TI - The malaise theory of depression: major depressive disorder is sickness behavior and antidepressants are analgesic. AB - The malaise theory of depression constitutes a re-conceptualization and re definition of major depressive disorder (MDD). It is proposed that the state or emotion of malaise should be considered the core symptom of depression, rather than sadness of mood. The syndrome of MDD is identified as a consequence of inappropriate sickness behavior mediated by immune activation including abnormalities in cytokines. Antidepressants are suggested to exert their specifically beneficial effects through an analgesic effect on the core dysphoric symptoms of malaise. These ideas are consistent with a substantial body of published literature and lead to a wide range of testable predictions. PMID- 10790738 TI - It is hazardous to treat HIV patients with interferon-alpha. AB - Interferon-alpha is shown to play a key role in the progressing immunodysfunction that characterizes HIV infection. In particular, interferon-alpha is responsible for the development of an autoimmune state that is prone to serious complications. Therefore, treatment of HIV patients with IFN-alpha is hazardous. There are results suggesting that anti-interferon-alpha immunization might be a method for prophylaxis and treatment of the autoimmune state in HIV patients. This treatment may prevent complications of HIV infection and the transition to AIDS. PMID- 10790739 TI - Association of sudden infant death syndrome with grossly deranged iron metabolism and nitric oxide overload. AB - Sudden infant death syndrome (SIDS) occurs silently usually during sleep and, though remaining unexplained after autopsy, leaves footprints creating a pattern analogous to that which follows a flood of nitric acid (NO). These footprints in SIDS are associated with serious pathological changes, viz. elevated hepatic iron, bone marrow hyperplasia, hypomyelinated respiratory control centres, elevated lung immunoglobulins, cerebral hypoperfusion resembling lesions induced by chronic hypoxemia, ischemia, congenital heart disease and congenital myopathy. Hypoxia stimulates the immune response and the over-arousal of the immune response triggers a flood of NO. Adenosine triggers sleep. NO and adenosine are additive as dilators of coronary blood vessels. Blood pressure collapses. Selenium increases the activity of the enzyme ferrochelatase during incorporation of heme into cytochrome oxidase. NO binds to cytochrome oxidase, inhibiting respiration. When NO reaches dangerous levels, the cell turns on production of heme oxygenase. Heme is broken down to iron (Fe) carbon monoxide (CO) and bile pigments. NO has a huge affinity for hemoglobin which catalyses NO degradation to nitrate. Furthermore, NO is a product of smoke and SIDS incidence is higher in smoking mothers. PMID- 10790740 TI - Urinary tract anomalies and dysfunctional voiding: a spectrum dictated by the influence of amniotic pressure upon fetal urodynamics. AB - The etiologic basis of urinary tract anomalies and dysfunctional voiding largely remain unknown. However, the travel of urine from renal pelvis into the amniotic cavity under the pressures exerted by amniotic fluid satisfactorily explains the etiologic basis. Amniotic pressure is affected by the changes in maternal intraabdominal pressure. The intraabdominal pressure of the fetus is also dictated by the amniotic pressure. Amniotic pressure compresses the urethra throughout the length, and may increase both bladder leak point pressure and urethral resistance. Furthermore, the urine is propelled against amniotic pressure. These factors closely simulate outflow obstruction. Since the pressure within the bladder with minimal urine content reflects the intraabdominal pressure, intravesical pressure is also elevated during fetal life. Additionally, elevated intravesical pressures impair ureteral drainage. The compressive effect exerted by fetal intraabdominal pressure upon ureters, further elevate the pressure within the renal pelvis. While forwarding the urine against these pressures may result in anomalies of the urinary tract, the increased work load of the detrusor may act as a state of injury that forms the basis of dysfunctional voiding. PMID- 10790741 TI - The toxic mind: the biology of mental illness and violence. AB - The continual suppression of emotions during fight or flight reactions results in atrophy and endogenous toxicosis in noradrenergic neurons. Toxic metabolites interfere with neurotransmission, causing depression. During periodic detoxification crises, excess norepinephrine floods synapses overexcite postsynaptic neurons, and cause symptoms ranging from mild anxiety to violent behavior. When toxic metabolites, which may include excess dopamine, epinephrine, serotonin, gamma-aminobutyric acid, peptides, amino acids, and various metabolic waste products, are bound to noradrenergic receptor sites, these sites become unavailable to norepinephrine. Excitation of postsynaptic neurons is diminished and depression returns. The diverse proteins in receptor sites presumed to be specific for false neurotransmitters may instead encode specific memories. The shift in depressive and excitatory behavior is characteristic of most psychiatric disorders, addictions, Alzheimer's disease, Parkinson's disease, and psychosomatic disorders. Recovery is a detoxification process and can be facilitated by therapy that involves re-experiencing childhood traumas, releasing, and redirecting repressed emotions. PMID- 10790742 TI - Schizophrenia: a purinergic hypothesis. AB - Knowledge of the physiological roles of the purinergic system and its influence on other neurotransmitter systems has greatly advanced. In this article, a purinergic model is proposed as an attempt to integrate several findings in schizophrenia. According to this hypothesis, a purinergic system dysfunction would mainly result in reduced adenosinergic activity. This model also addresses the systemic aspects of schizophrenia, based on peripheral roles of purines, such as modulation of the immune system. PMID- 10790743 TI - Reading for empathy. AB - As our preoccupation with empathy grows, we cast about for means of inculcating it. Literature has been proposed as an enabling factor for physicians in developing a creative imagination. This essay, using a number of examples, points out that reading for pleasure in areas not necessarily related to medicine, can yield insights into its practice. Thus, sensitizing students and young physicians to the nuances of human interaction described by great writers could constitute part of basic and continuing medical education and might be one way of instilling empathy. PMID- 10790744 TI - 'Receptor component' and 'active component' in the psychology and psychopathology of perception. AB - Sensory perception should not be regarded as one starting only (and solely) when the receptor is being stimulated. Along with the receptor component, the active component plays an essential role in the perception process. The main elements forming the active component are: expectation of a certain signal, prediction of the signal most probable in this particular situation, and an affective distinction determining the necessity of the signal for the individual perceiving the signal. Hence, a sequence of sensory phenomena can be formed (both for the normal and pathological states) depending on the relation between the receptor and active components. PMID- 10790745 TI - An interesting observation of the position of some electronegative atoms in the taxol molecule. AB - A study of the absolute energy-minimized configuration of the taxol molecule revealed that many oxygen atoms in this molecule and a nitrogen atom on the side chain are located on the same plane. A comparison of these electronegative atoms with information provided from existing literature indicated that most of these noted atoms conform with the reported structure--activity relationship (cytotoxicity, antineoplastic activity, tubulin assembly property, etc.). This observation may assist investigators in their future design of simpler, possibly clinically efficacious molecules related to taxol. PMID- 10790746 TI - Selective treatment of neoplastic cells using ferritin-mediated electromagnetic hyperthermia. AB - A new method of cancer treatment is proposed, based on the unique magnetic properties of ferritin iron core which, in alternating magnetic field of frequency approximately 100 kHz, is easily heated to temperatures sufficiently high to destroy neoplastic cells containing an excess of this protein, without damaging the normal cells. PMID- 10790747 TI - Gene transfer-mediated intracellular photodynamic therapy. AB - The main limitation of photodynamic therapy is a very short penetrance length of the light in tissues. To overcome this shortcoming, a new method is proposed, where first a gene encoding protein luciferase is delivered using, e.g. adenovirus vector to the neoplastic cells and, after its expression, the photosensitizer, which should be activated, together with luciferin are injected to the organism. After their accumulation in the cancer cells, the photosensitizer activation is accomplished via light produced by chemiluminiscent reaction of luciferase and luciferin. PMID- 10790748 TI - Obesity and osteoarthritis of the knee: hypotheses concerning the relationship between ground reaction forces and quadriceps fatigue in long-duration walking. AB - The principal risk factors of osteoarthritis of the knee are: age, obesity and gender. It is hypothesized that long-duration walking (e. g. 20 min) in the elderly obese will lead to quadriceps fatigue. Changes in the gait pattern due to fatigue will lead to altered knee kinematics at heelstrike and consequently decreased shock absorption. This scenario will result in an increased rate of loading and possibly an increase in the overall magnitude of peak ground reaction forces, both of which could cause articular cartilage degeneration. Obese females are at an overall higher risk of developing osteoarthritis than males. This gender discrepancy may be explained by the fact that females have a higher percentage of body fat content (lower proportion of lean mass) that may increase the rate of quadriceps fatigue. These biomechanical hypotheses can be examined by studying continuous periods of walking in which ground reaction forces, knee kinematics and electromyography data are recorded. PMID- 10790749 TI - Transmissible spongiform encephalopathies and the 'rogue share-holder protein' hypothesis. AB - Transmissible spongiform encephalopathies(TSEs) or prion diseases are a closely related group of diseases, whose exact etiology is unknown, but is generally accepted to be related to protease-resistant prion protein PrP. PrPc is normally present in cells and its disease counterpart PrPsc is postulated to occur due to a rare stochastic change. The selfish gene hypothesis is a generally well accepted concept in evolutionary biology. Genes can be likened to the board of a company and proteins can be likened to share-holders. Here it is being hypothesized that a rogue share-holder protein's 'selfish' replicatory tendency might be the explanation for TSEs. The present hypothesis predicts existence of other examples of rogue share-holder protein and also predicts that examples would be found in lower life-forms as well. PMID- 10790750 TI - Is mitral valve prolapse a manifestation of adolescent growth spurt? AB - The exact aetiology of mitral valve prolapse (MVP) is unknown, although this is the most common cardiac valvular abnormality currently detected. MVP has high incidence in young individuals, particularly during the second and third decades. These individuals are usually of a slender body habitus indicating higher rates of linear growth, reflective of the adolescent growth spurt. MVP might represent the imbalance in the growth dynamics of the mitral valve apparatus especially between the leaflets, chordae tendineae and the rest of the heart. Several reports suggest the transient nature of MVP and even complete disappearance. MVP with systolic click, but without a systolic murmur signifying regurgitation may be considered as a manifestation of adolescent growth spurt and a normal variant transiently occurring during particular periods of lives of otherwise normal individuals. Strategies of identification of subsets of individuals likely to harbor the more sinister and progressive form of MVP are important and need to be developed. PMID- 10790751 TI - Cell phenotype as a target of drug therapy in chronic inflammatory diseases. AB - Many diseases share common pathological changes which could in principle be targets for new drugs. Vascular leakage of plasma and migration of cells into perivascular tissues are common to chronic inflammatory diseases such as asthma, atherosclerosis, arthritis, and proliferative nephropathy as well as some non inflammatory proliferative disorders such as diabetes mellitus. Individual components of plasma have been shown to stimulate cellular proliferation, matrix deposition and phenotypic change, leading to tissue-damaging structural changes. Whereas most anti-inflammatory drugs either downregulate expression of inflammatory mediators or inhibit their actions on cells, there are alternate potential therapeutic strategies described here based on moderating vascular leakage or its consequences in chronic diseases. The hypothesis is that drugs that can modify a cell's phenotype could be used to limit structural changes which accompany inflammation and thus reduce permanent debility resulting from these diseases. Such drugs include the differentiating agents being developed for cancer therapy. PMID- 10790752 TI - Microglia dysfunction in schizophrenia: an integrative theory. AB - Schizophrenia is a devastating illness of unknown etiology. It is characterized by increased brain ventricular volume, suggesting a progressive neurodevelopmental condition. There is evidence suggesting a correlation between in utero viral exposure and subsequent occurrence of schizophrenia. Many neurotransmitter systems have been implicated as being dysfunctional in schizophrenia. There are also data suggesting immune system dysfunction in schizophrenia, and a negative correlation between schizophrenia and rheumatoid arthritis. Microglia are phagocytic immune cells in the central nervous system (CNS) derived from peripheral blood monocytes. They are involved in brain development, neuroproliferative and neurodegenerative activities, several CNS illnesses, and CNS viral immunity. They may also be involved in neurotransmitter regulation. The current theory postulates microglial dysfunction initiated by early CNS viral exposure results in the abnormal neural development and neurotransmitter dysfunction seen in schizophrenia. PMID- 10790753 TI - The 'rotational preconstraint'. AB - This model provides a novel view of the etiology of some scolioses and can answer some of the biomechanical questions regarding pathogenesis of dorsal curves. According to our findings, paravertebral muscular imbalance is likely to favour such a pathological condition which, with the interference of the postural reflexes and the body weight-related vertical loading, might lead to the formation of a true scoliotic curve. Review of earlier research studies in the light of our findings reveals controversy in some authors' reported results and their own interpretations and seems to generally support our theory. PMID- 10790754 TI - Psychiatric diagnoses and inconsistent results of association studies in behavioral genetics. AB - Case-control association studies investigating polymorphisms of candidate genes in behavioral disorders have produced a lot of positive and negative findings with few consistent replications. The inconsistent results of association studies in behavioral genetics are usually explained by the fact that significant appearing relationships may be found as an artifact of genetic differences between the cases and controls because population stratification (or admixture) due to ethnic variation or other confounding factors can generate considerable population differences in marker allele frequencies. The author suggests that a major problem of association studies in psychiatric genetics is that psychiatric diagnoses are not biologically real disease entities: syndromal psychiatric diagnostic categories such as depression or schizophrenia include etiologically, pathologically, and prognostically heterogenous disorders. The author further suggests that extensive clinical research and observations are necessary to classify heterogeneous diagnostic categories into more scientific and more homogenous disease entities. PMID- 10790756 TI - Oral vaccine in otosclerosis. AB - Otosclerosis is a genetically based disease in which the development of autoimmune mechanisms can lead to its clinical expression. Anti-type II collagen antibodies have been found in otosclerotic patient serum. The first clinically unexpressive phase of the disease can be diagnosed from a family history and ON OFF stapedial reflexes with evidence of an autoimmune disorder. The use of antigenic oral hyposensitization is accepted, based on autoimmune pathogenesis. Hyposensitization should be initiated during this subclinical period of otosclerosis with a type II collagen oral vaccine. PMID- 10790755 TI - Is inhibition of cyclooxygenase required for the chemopreventive effect of NSAIDs in colon cancer? A model reconciling the current contradiction. AB - NSAIDs are powerful chemopreventive agents for colon cancer, but their mechanism of action remains unknown. Their best recognized pharmacological property is inhibition of the enzyme cyclooxygenase (COX), which catalyzes the synthesis of prostaglandins; however, additional effects are well documented. Current studies on the mechanism of the chemopreventive effect of NSAIDs lead to two contradictory conclusions: NSAIDs prevent colon cancer either by inhibiting the activity of COX, or through mechanisms that do not require COX inhibition. To resolve this apparent conflict, after examining several alternatives, we propose a model, which assumes that both mechanisms are correct but that they exert their effect either on different steps of the multistep process of colon carcinogenesis or on different control mechanisms. This postulated dual action of NSAIDs may explain their remarkable effectiveness in colon cancer prevention. Unraveling these mechanistic details can be very rewarding for the design of more refined approaches to cancer chemoprevention and for a deeper understanding of colorectal carcinogenesis. PMID- 10790757 TI - Potential role for Ginkgo biloba extract in the treatment of glaucoma. AB - Glaucoma is becoming recognized as a condition for which not only elevated intraocular pressure, but also non-pressure-dependent risk factors are responsible. New avenues of treatment into which investigations are being initiated include agents which could possibly improve blood flow to the eye and neuroprotective drugs. Only calcium channel blockers are presently available for such treatment in glaucoma, and these have not been widely adopted, in contrast to clinical trials involving a number of neuroprotectants in other neurologic disorders. Ginkgo biloba extract is freely available and has several biological actions which combine to make it a potentially important agent in the treatment of glaucoma: improvement of central and peripheral blood flow, reduction of vasospasm, reduction of serum viscosity, antioxidant activity, platelet activating factor inhibitory activity, inhibition of apoptosis, and inhibition of excitotoxicity. The effect of Ginkgo biloba extract as a potential antiglaucoma therapy deserves intensive scrutiny. PMID- 10790759 TI - On the distribution of pivots among C units of mammalian chromatids. AB - The paper discusses how a DNA strand contributes pivots that distributively interconnect the jaws of two or more C units of a chromatid stack. Rather than being static, the location of these pivots may be shifted according to alternative modes without compromising the chromatid's physical integrity. PMID- 10790758 TI - Pathogenesis of anaemia in hyperparathyroidism. AB - It is suggested that parathyroid hormone (PTH), when in excessive amounts, interferes with normal erythropoiesis by downregulating the erythropoietin receptors on erythroid progenitor cells in the bone marrow. Therefore, physiologic concentrations of EPO can no longer sustain normal red cell counts, so normocytic and normochromic anaemia ensues. In primary hyperparathyroidism (HPT), this effect is observed with very high concentrations of PTH. In secondary HPT during chronic renal failure, this effect is more pronounced because erythropoietin synthesis is impaired. PMID- 10790760 TI - Cardiac afferents and neurohormonal activation in congestive heart failure. AB - Cardiac chambers have afferent connections to the brainstem and to the spinal cord. Vagal afferents mediate depressor responses and become activated by volume expansion, increased myocardial contractility and atrial natriuretic factor. Sympathetic afferents, on the contrary, are activated by metabolic mediators, myocardial ischemia and cardiac enlargement. These opposite behaviors may lead to activation or suppression of the sympathetic nervous system and of the renin angiotensin-aldosterone system. As cardiac diseases progress, the heart dilates, plasma norepinephrine increases, atrial natriuretic factor is released and the renin-angiotensin-aldosterone system is suppressed to maintain water and sodium excretion. This dissociation of the neurohormonal profile of cardiac patients, may be explained by coactivation of sympathetic afferents, by cardiac dilatation, and of vagal afferents by atrial natriuretic factor. In more advanced stages, atrial natriuretic factor suppression of the renin-angiotensin-aldosterone system is overridden by overt sympathetic activation and sodium and water retention ensues. Digitalis, angiotensin-converting enzyme inhibitors and beta-blockers selectively decrease cardiac adrenergic drive. A common mechanism of action, to all three groups of drugs, would be attenuation of sympathetic afferents and partial normalization of vagal afferents. Consequently, heart size and cardiac afferents emerge as the key factors to understand the pathophysiology and treatment of the syndrome of congestive heart failure. PMID- 10790761 TI - Effects of food processing on the thermodynamic and nutritive value of foods: literature and database survey. AB - One of the goals of our society is to provide adequate nourishment for the general population of humans. In the strictness sense, the foodstuffs which we ingest are bundles of thermodynamic energy. In our post-industrial society, food producers provide society with the bioenergetic content of foods, while stabilizing the food in a non-perishable form that enables the consumer to access foods that are convenient and nutritious. As our modern society developed, the processing of foodstuffs increased to allow consumers flexibility in their choice in which foods to eat (based on nutritional content and amount of post-harvest processing). The thermodynamic energy content of foodstuffs is well documented in the literature by the use of bomb calorimetry measurements. Here, we determine the effects of processing (in most cases by the application of heat) on the thermodynamic energy content of foods in order to investigate the role of processing in daily nutritional needs. We also examine which processing procedures affect the nutritive quality (vitamin and mineral content) and critically assess the rational, advantages and disadvantages of additives to food. Finally, we discuss the role of endogenous enzymes in foods not only on the nutritive quality of the food but also on the freshness and flavor of the food. Our results show that a significant decrease in thermodynamic energy content occurs in fruits, vegetables, and meat products upon processing that is independent of water content. No significant change in energy content was observed in cereals, sugars, grains, fats and oils, and nuts. The vitamin content of most foods was most dramatically decreased by canning while smaller effects were observed upon blanching and freezing. We found that most food additives had very little effect on thermodynamic energy content due to their presence in minute quantities and that most were added to preserve the foodstuff or supplement its vitamin content. The endogenous food enzymes while aiding in digestibility of some foods (yogurt or grains) also helped some foods have a more palatable taste. Our conclusions are there is some scientific merit to the idea that enzymes in food can act synergistically with those in the human body to facilitate maximum nutritive value of foods. PMID- 10790762 TI - NF-kappaB, nitric oxide and opiate signaling. AB - NF-kappaB, a DNA binding factor, has been implicated in inflammatory cytokine activation. NF-kappaB is activated by IkappaBalpha, its inhibitor, which is phosphorylated and proteolytically degraded. In this regard, NF-kappaB is also responsive to reactive oxygen intermediates and calcium. Reports also have emerged that demonstrate that nitric oxide inhibits NF-kappaB transcriptional activation in a variety of cells, including monocytes and endothelial cells. Recently, we have demonstrated that morphine, not opioid peptides, via the mu3 opiate receptor is coupled to constitutive nitric oxide release in these same cells. In this regard, we provide a scenario whereby morphine modulates NF-kappaB activation via nitric oxide. This pathway appears to be the key step in regulating inducible nitric oxide synthase expression, controlling the balance between constitutive nitric oxide synthase and the inducible form. PMID- 10790763 TI - Molecular mechanism of monoamine toxicity in Parkinson's disease: hypothetical cell death model. AB - Although there have been experimental approaches to understanding the etiology of Parkinson's disease, the cause of cell degeneration in this neurological disorder remains a mystery. Herein, a hypothetical model is proposed to explain the mechanism leading neurons to die. The model is based on recent experimental evidence and it attempts to dissect the actions of dopamine and metal ions as potential triggers for the activation of an ordered cascade of events of the cell death machinery. PMID- 10790764 TI - Helicobacter infection and cirrhosis in hepatitis C virus carriage: is it an innocent bystander or a troublemaker? AB - Since it has been shown that Helicobacter hepaticus causes both chronic hepatitis and hepatocellular carcinoma (HCC) in mice, it is suggested that differences in the progression of chronic hepatitis C may be due to a cofactor stemming from co infection by bacteria, especially Helicobacter pylori, and/or other Helicobacter species. An assessment was made of the prevalence of H. pylori infection in HCV positive cirrhotic patients. The presence of Helicobacter species (spp). was evaluated in resected liver tissue from HCC patients. Serum anti-H. pylori IgG antibodies were determined in 70 males with a clinical and/or histological diagnosis of cirrhosis and HCV infection and in 310 age-matched male blood donors. The prevalences of H. pylori antibody were 77% (54/70) and 59% (183/310) (P 0.004). Primers identifying 26 Helicobacter species were used to determine the presence of the genomic 16S rRNA of this genus in liver tissue resected from 25 cirrhotic HCC patients. Genomic sequences corresponding to H. pylori and H. pullorum were identified in 23 of these 25 livers. Together, these findings support the proposal that H. pylori is implicated in the pathogenesis and progression of cirrhosis, particularly in HCV-infected individuals. Involvement of Helicobacter spp. in HCC also seems highly possible. PMID- 10790766 TI - Tachycardia: an autosomal, monogenic, biallelic, recessive trait. AB - A brief commentary of the genetics of blood pressure is presented. The importance of the mechanisms of blood pressure regulation, among which heart rate is relevant, is emphasized. The analysis of a small population to test the quantitative model of the trait heart rate, considered as a metric character, is presented. The analysis of heart rate fitting to a qualitative model of inheritance is carried out. The results displayed might support the hypothesis that tachycardia could be an autosomal, monogenic, biallelic recessive trait. PMID- 10790765 TI - Ecosystems supporting clusters of sporadic TSEs demonstrate excesses of the radical-generating divalent cation manganese and deficiencies of antioxidant co factors Cu, Se, Fe, Zn. Does a foreign cation substitution at prion protein's Cu domain initiate TSE? AB - Analyses of food chains supporting isolated clusters of sporadic TSEs (CWD in N Colorado, scrapie in Iceland, CJD in Slovakia) demonstrate a consistent 2 1/2+ fold greater concentration of the pro-oxidant divalent cation, manganese (Mn), in relation to normal levels recorded in adjoining TSE-free localities. Deficiencies of the antioxidant co factors Cu/Se/Zn/Fe and Mg, P and Na were also consistently recorded in TSE foodchains. Similarities between the clinical/pathological profile of TSEs and Mn delayed psycho-neurotoxicity in miners are cited, and a novel theory generated which suggests that sporadic TSE results from early life dependence of TSE susceptible genotypes on ecosystems characterised by this specific pattern of mineral imbalance. Low Cu/Fe induces an excessive absorption of Mn in ruminants and an increased oxidation of Mn2+ into its pro oxidant species, Mn3+, which accumulates in mitochondria of CNS astrocytes in Mn SOD deficient genotypes. Deficiencies of scavenger co factors Cu/Zn/Se/Fe in the CNS permits Mn3+ initiated chain reactions of auto-oxidant mediated neuronal degeneration to proliferate, which, in turn, up-regulates the expression of the Cu-metalloprotein, prion protein (PrP). Once the rate of PrP turnover and its demand for Cu exceeds the already depleted supply of Cu within the CNS, PrP can no longer bind sufficient Cu to maintain its conformation. Mn3+ substitutes at the vacated Cu domain on PrP, thus priming up a latent capacity for lethal auto oxidative activity to be carried along with PrP like a 'trojan horse'; where Mn 3+ serves as the integral 'infectious' transmissible component of the misfolded PrP-cation complex. The Mn overactivation of concanavalin A binding to glycoprotein and Mn-initiated autoxidation results in a diverse pathological profile involving receptor capping, aggregation/modification of CNS membrane/cytoskeletal proteins. TSE ensues. The BSE/nv CJD strain entails a 'synthetic' induction of the same CNS mineral disturbance, where 'in utero' exposure to Cu-chelating insecticides/Mn supplements accelerates the onset of a more virulent 'strain' of adolescent TSE. PMID- 10790767 TI - Diseases of plants transmissible between plants and man (Phytonoses) exist- follow-up paper. AB - The purpose of this paper is to revise the original paper by qualifying a statement made therein and making two sections more inclusive. The qualification is the insertion of the word 'adult' before the word 'volunteers' in one sentence. The scope of the first section is increased by: (i) drawing attention to the possibility that besides other organisms (i.e. non-mycoplasma-like organisms (non-MLOs)) and viruses that could be inadvertently transmitted between members of the plant and animal kingdoms, other infectious entities including viroids and prions and/or prion-like elements should also be considered in this regard; and (ii) the need to consider the possible transmission of either antibiotic-resistant MLOs and/or MLOs harbouring entities that are either naturally infectious and pathogenic for man or may opportunistically infect man. The scope of the second section is increased by proposing additional possible mechanisms of transmission of MLOs between plants and man. PMID- 10790768 TI - Theoretical mechanistic basis of oxidants of methaemoglobin formation. AB - The mechanism of oxidation or reduction using the electron method was investigated for (I) aniline; (II) nitrobenzene; (III) nitrate; (IV) sulphanilamide; (V) hydrogen peroxide; (VI) hydroxyl free radical; (VII) ferricyanide; (VIII) acetylphenylhydrazine; (IX) nitrite; (X) chlorate and (XI) hydroxylamine respectively. Substances (II), (III), (V), (VI), (VII), (IX), (X) and (XI) evolved as oxidants, with (II), nitrobenzene and (X), chlorate as the most powerful oxidants (number of moles of HbFe(2+)(haemoglobin) of 6 reacting with 1.0 mole of the substance). Substances (I), (IV) and (VII) evolved as reductants of equal reducing power (number of moles of HbFe(3+)(methaemoglobin) of 4 reacting with 1.0 mole of the substance). Using the following equations, the impact of oxidants and reductants on glutathione (GSH) peroxidase, glutathione (GSSC) reductase and NADHmetHb reductase respectively on methaemoglobinaemia generation was investigated. [Equation in text]. Redox potential change (DeltaE' (o)) of 1.77, -1.77 and 1.86 volt and free energy change (DeltaG(o)') of -81, 81 and -85.8 kcal/mol were calculated for GSH peroxidase, GSSG reductase and NADHmetHb reductase systems respectively. In sustained methaemoglobinaemia, these mechanisms predict low levels of NADHmetHb reductase and glutathione peroxidase respectively, but high levels of glutathione reductase in red blood cells on exposure to oxidants. The significance of these mechanisms was investigated in cord blood, neonatal, adult red blood cells and other biological systems. It was concluded that any reaction with a positive DeltaE(o)' and negative DeltaG(o)' with the Fe(3+): Fe(2+)couple will indicate methaemoglobin oxidizing power. The effects on red blood cells and white blood cells were manifested in the biochemical toxicology of nitroso (PhN = 0), arylamine glucuronide (PhNHG) and arene imine respectively. PMID- 10790769 TI - Etiology of multiple sclerosis: role of superoxide dismutase. AB - The etiology of multiple sclerosis is still unknown, despite intensive research for many decades. It has, however, been shown recently that defects in the genes may be responsible for conditions such as amyotrophic lateral sclerosis (1). PMID- 10790770 TI - Sublethal oxygen deficiency--the primary cause of cancer. PMID- 10790771 TI - Shattered: Medawar's test tubes and their enduring legacy of chaos. AB - Medawar's 1952 paper 'An Unsolved Problem of Biology' underlies most subsequent theoretical work regarding the evolution of aging; it concludes that aging is accidental and could not have evolved; this prevents reconciling the growing body of evidence suggesting the existence of multiple, evolved, aging systems. The paper features a well-known thought experiment using test tubes to show why aging could not evolve. Medawar assumes that constant, random, breakage sufficiently represents lethal forces of nature; however, famine, drought, predation, disease, and accidents each uniquely affect populations. Predation is the only evolving force that continually invents new ways to kill members of the prey populations; thus all prey defenses to predation will eventually be defeated. Defenses to non evolving or non-obligate lethal forces, however, should quickly evolve. Thus unevolving, identical test tubes cannot adequately represent biological populations. The example also ignores population booms and busts which often occur in nature. By ignoring these issues, Medawar examines only one population age distribution skewed towards younger individuals in predator-dominated environments while ignoring predator-free populations skewed towards older individuals after population crashes. Further, Medawar's test tubes lack meaningful competition for finite resources, and ignore declining fertility which occurs in all aging species. Medawar concludes that older individuals are too few in number to influence the population's gene pool for or against aging. This conclusion is found to be incorrect when variations in the age of reproductive senescence are introduced into a predator-free population.A new thought experiment with competing strains of algae corrects for these issues and shows that aging evolved and is retained so that groups retain enough genetic variability to allow for rapid evolution of a defense to novel predation. The example shows reasons why the rate of aging is directly linked to the reproductive rate, litter size, metabolic rate, reproductive senescence, and fixed body size. It also suggests that in the absence of predation, immortality would quickly evolve if not for the evolution of highly-conserved aging systems. Prior analysis of aging evolution is incorrect due to theorists' rejection of the idea of group selection. It is believed to be 'impossible' to select for mutations that are bad for the individual but good for the group. However, mutations that are neutral to young individuals which are only deleterious if expressed at older ages can accumulate in early-mortality, predator-dominated environments. Removing the predator allows deleterious mutation expression. Positive group selection then occurs amongst traits that are negative to the individual. Further, group selection is a universal force that occurs between local, non-breeding groups and not, as theorists propose, between distant groups of potentially interbreeding species. Local survivors migrate to replace extinct, related species. The antagonistic pleiotropy theory, which was created to salvage the idea of accidental aging, is examined and shown to be untenable. The hypothetical antagonistic pleiotropy genes that are beneficial to young while detrimental to old individuals, predicted to exist in the 1950s, are unlikely to exist, have not, and likely will not be found in sufficient quantity to participate in the aging process. PMID- 10790772 TI - Short note: a novel approach for treating early-stage acute leukemias. AB - A novel approach to treating acute leukemias is suggested by hypothesizing a focal unique origin in a focal area of bone marrow and this area can be localized (and ablated) before the leukemia stem cells have seeded other bone marrows. PMID- 10790773 TI - Melatonin, psoralens, and the genetic basis of seasonal affective disorder and seasonality. PMID- 10790774 TI - Electrochemical treatment of tumours. AB - The electrochemical treatment (EChT) of tumours implies that tumour tissue is treated with a continuous direct current through two or more electrodes placed in or near the tumour. The treatment offers considerable promise of a safe, simple and relatively noninvasive anti-tumour therapy for treatment of localised malignant as well as benign tumours. Although more than 10,000 patients have been treated in China during the past 10 years, EChT has not yet been universally accepted. The reason for this is the lack of essential preclinical studies and controlled clinical trials. Uncertainties regarding the destruction mechanism of EChT also hinder the development of an optimised and reliable dose-planning methodology. This article reviews the collected Chinese and occidental experiences of the electrochemical treatment of tumours, alone and in combination with other therapies. The current knowledge of the destruction mechanism underlying EChT is presented along with different approaches towards a dose planning methodology. In addition, we discuss our view of different important parameters that have to be accounted for, if clinical trials are to be initiated outside of China. PMID- 10790775 TI - Non-linear dielectric spectroscopy: antifouling and stabilisation of electrodes by a polymer coating. AB - Non-linear dielectric spectroscopy (NLDS) has previously been shown to produce quantitative information that is indicative of the metabolic state of various organisms, by modeling the non-linear effects of their membranous enzymes on an applied oscillating electromagnetic field using supervised multivariate analysis methods. However, the instability of the characteristics of the measuring apparatus rendered the process temperamental at best in the laboratory and impractical for field use. The main practical problem, of the non-stationarity of the electrode-solution interface and the ease with which the electrode surfaces are subject to protein fouling. It is addressed by applying a thin, electrically transparent antifouling coat to the electrodes. This reduces the interminable cleaning procedures previously required to prepare the electrodes for use, increases their usable lifetime before recleaning, and also improves the precision and linearity of multivariate models on NLDS data. PMID- 10790777 TI - Measurement of proton release and uptake by analogs of bacteriorhodopsin. AB - Proton release and subsequent uptake by several forms of bacteriorhodopsin (bR), including 4-keto analogs of wild-type (WT) and D96N and D85N mutants as well as the 9-demethylretinal analog of WT and D96N mutants, have been measured using a highly sensitive electrochemical technique. Release and uptake of protons by bR in membrane patches on a tin oxide electrode produce a current transient whose amplitude is proportional to the rate of pH change at the electrode surface. Profiles of proton release by the analogs vs. pH are substantially different from the profiles of the native proteins. PMID- 10790776 TI - Attachment of bacterial cells to carbon electrodes. AB - Anodic stripping method was applied to analyze the process of bacterial attachment to the surface of carbon-paste electrodes (CPE). The electrode was immersed for various times in a bacterial cell suspension to allow the cells to attach to its surface. The number of bacterial cells attached to the electrode surface increased along with time. On the other hand, the current derived from the oxidation of a dye, Hoechst, which was adsorbed to the surface after attaching the bacterial cells, decreased along with time. It was considered that the current output, correlated with the amount of dye, adsorbed onto regions where no bacterial cell attached. These results indicate that the bacterial attachment process can be analyzed by measuring the electric current derived from the dye instead of counting the number of attached cells. PMID- 10790778 TI - Electrochemical study of the effect of ADP and AMP on the kinetics of glutamate dehydrogenase. AB - A chronoamperometric method based on the 'diffusion' layer concept of the convective system was used to assay the glutamate dehydrogenase (GLDH) activity. Once the reaction was initiated by adding the enzyme GLDH into a well-stirred nicotinamide adenine dinucleotide (NADH, coenzyme) solution, the steady-state oxidation limiting current of NADH would decrease linearly in a short time. The major advantage of this method is that it directly indicates the continuous in situ change of the coenzyme concentration, thus, the real initial reaction rate of enzyme-catalyzed reaction, V0, can be determined. Using this method, the effect of adenosine-5'-monophosphate (AMP) and adenosine-5'-diphosphate (ADP) on the GLDH activity has been monitored. The results showed that ADP and AMP could increase the activity of GLDH. This activation mechanism was proposed by the voltammetric study. PMID- 10790779 TI - Perturbation of human skin due to application of high voltage. AB - Electroporation is believed to be the effect that greatly enhances the transport of water-soluble molecules across the stratum corneum (SC) by application of short high voltage pulses. However, electroporation was originally a phenomenon investigated at the level of cell and model membranes, which is only partially comparable to the complicated structure of the stratum corneum. Here, we show, that electroporation is accompanied by other effects, which may be primarily involved in creation of new pathways and altering existing pathways, respectively. Experimental evidence shows that the dramatic increase in skin permeability is due to synergistic effect of electric field and heating by high local current density. Heating starts at small spots, not related to a visible skin structure and results in a propagating heat front. The phase transition of the SC lipids plays a major role in skin permeability during the pulse. The permeability after a high voltage pulse correlates well with the surface area showing a permanent low electrical resistance after pulsing. The main transport of water-soluble molecules is facilitated by the electric field due to the electrophoretic driving force in conjunction with the high permeability due to the breakdown of the multilamellar system of the SC lipids. PMID- 10790780 TI - Correction of the influence of baseline artefacts and electrode polarisation on dielectric spectra. AB - The deconvolution of biological dielectric spectra can be difficult enough with artefact-free spectra but is more problematic when machine baseline artefacts and electrode polarisation are present as well. In addition, these two sources of anomalies can be responsible for significant interference with dielectric biomass measurements made using one- or two-spot frequencies. The aim of this paper is to develop mathematical models of baseline artefacts and electrode polarisation which can be used to remove these anomalies from dielectric spectra in a way that can be easily implemented on-line and in real-time on the Biomass Monitor (BM). We show that both artefacts can be successfully removed in solutions of organic and inorganic ions; in animal cell and microbial culture media; and in yeast suspensions of varying biomass. The high quality of the compensations achieved were independent of whether gold and platinum electrodes were used; the electrode geometry; electrode fouling; current density; the type of BM; and of whether electrolytic cleaning pulses had been applied. In addition, the calibration experiments required could be done off-line using a simple aqueous KCl dilution series with the calibration constants being automatically calculated by a computer without the need for user intervention. The calibration values remained valid for a minimum of 3 months for the baseline model and indefinitely for the electrode polarisation one. Importantly, application of baseline correction prior to polarisation correction allowed the latter's application to the whole conductance range of the BM. These techniques are therefore exceptionally convenient to use under practical conditions. PMID- 10790781 TI - Electric-field effects in dry films of D85N and D85,96N mutant bacteriorhodopsin. AB - In the D85N mutant of the protein bacteriorhodopsin (BR), the Schiff base, by which the retinal chromophore is bound to the protein, exhibits an abnormally low proton affinity (pKa approximately 8.9). Recent experiments on thin films of this protein have shown that this causes the protonation state of the Schiff base, and thus the visible absorption spectrum, to be sensitive to external electric fields. In this paper, we explore the dependence of this effect on parameters such as pH, humidity, and film thickness. The results of these experiments point to the importance of water molecules bound in the acceptor part of the proton channel as sources and donors in field-induced proton-transfer reactions. We describe additional results obtained with the D85,96N mutant, which also exhibits a low Schiff-base pK. The similar behavior of the two mutants under applied electric fields at high pH implies that the residue Asp-96 plays no role in field induced Schiff-base protonation. PMID- 10790782 TI - Overcoming electrically induced artifacts in penetration studies with fluorescent tracers. AB - The use of model molecules in transdermal transport studies reveals transport behavior while providing an economical setup by detection of readily measured quantities such as fluorescence, radioactivity or absorbance of low-cost substances. Water soluble fluorescent tracers such as calcein have been repeatedly used as model molecules in transdermal transport studies. However, if electrically enhanced calcein transport across the human skin barrier is measured, artifacts due to interaction between calcein and electrode byproducts influence the result. Here, we describe an experimental setup which avoids known artifacts and makes calcein or other fluorescent tracers a suitable model for transdermal transport studies. PMID- 10790783 TI - Metastatic alveolar rhabdomyosarcoma with increased serum creatine kinase MB and cardiac troponin T and normal cardiac troponin I. PMID- 10790784 TI - Revolutionizing health: communication can make a difference. PMID- 10790785 TI - Impact of the integrated Radio Communication Project in Nepal, 1994-1997. AB - The Radio Communication Project (RCP) in Nepal is an ongoing, theory-based, multimedia reproductive health campaign which began in 1995. It consists of two entertainment-education radio serials (a soap opera for the general public and a dramatized distance education serial for health workers), additional radio spot advertisements and promotions, and complementary print materials. This paper examines impact data from a variety of sources, including a pre- and postpanel survey of currently married women (N = 1905), three waves of clinic-based observations of client-provider interactions (N = 240 per wave) and client exit interviews (N = 240 per wave), and 2 years of clinic service statistics, in order to draw inferences about the separate and combined effects of the RCP components. The study found increased health worker interpersonal interaction skills, improved quality of client-provider interactions, increased client self-efficacy in dealing with health workers, improved client attitudes toward health services and toward the practice of family planning, increased adoption of family planning, and increased family planning service utilization, all attributable to the RCP. The panel data allowed statistical control of the influence of predisposing factors before the campaign on postcampaign ideation and behavior. The effect of the RCP on contraceptive behavior was largely indirect through its influence on ideation. Implications for the design of integrated, multimedia, entertainment-education campaigns and integrated evaluation designs are discussed. PMID- 10790786 TI - Power distance and collectivist/individualist strategies in alcohol warnings: effects by gender and ethnicity. AB - This research examined the responses of 73 Mexican American and Anglo young adults to four televised drinking-and-driving warnings. Warnings were manipulated into collectivist (emphasizing risks to family and friends) and individualist (emphasizing risks to self) appeals, and into high and low power distance appeals by attributing or not attributing warnings to the Surgeon General. Females rated the collectivist warnings, and males the individualist warnings, more believable. Respondents on average responded to the collectivist warnings most positively, regardless of gender or ethnicity. Anglos rated warnings without the Surgeon General as the source more believable than warnings with the Surgeon General as the source; the opposite was true for Latinos. Other interactions were also found. PMID- 10790787 TI - Testing different formats for communicating colorectal cancer risk. AB - This study assessed the extent to which different formats of informing men and women age 50 and over of the risks of colorectal cancer (CRC) affected their perceptions of their absolute and comparative (self versus other) 10-year and lifetime risks; emotional reactions about getting CRC; and screening intentions. Forty-four men and 78 women received information about the absolute lifetime risk of getting CRC. In addition, participants either did or did not receive information about (1) lifetime risk of getting CRC compared with other cancers, and (2) risk factors for CRC (age and polyps). Participants who received risk factors information were more likely to increase their perceived absolute 10-year and lifetime risks of getting CRC compared with participants who did not receive risk factors information. In addition, participants who received risk factors information were more likely to believe age was related to getting CRC and felt at greater risk for having polyps compared with participants who did not receive this information. None of the experimental conditions affected how worried, anxious, and fearful participants felt about getting CRC, nor did they affect screening intentions. Independent of experimental condition, participants tended to increase their intentions to get screened for CRC in the next year or two. Intention to be screened was more pronounced among participants who had been screened via a fecal occult blood test (FOBT) or sigmoidoscopy (SIG). Implications for the design of interventions involving the communication of CRC risks are discussed. PMID- 10790788 TI - An assessment of the health communication job market across multiple types of organizations. AB - This study seeks to answer three questions: (1) What is the employment outlook for health communication practitioners? (2) What specialized knowledge and skills should a competent health communication practitioner possess? and (3) How much academic training or professional experience is necessary to become a competent health communication practitioner? To this end, 104 employers of health communication practitioners, representing different types of large, medium, and small companies and organizations from various regions of the United States, were interviewed by telephone. The interview protocol was based on nine core health communication responsibilities identified by a working group of health communication academicians and practitioners. The study suggests a positive employment outlook, where those seeking jobs in health communication before the year 2000 could enjoy varied job opportunities in the wake of an anticipated moderate expansion in the field. Those with 1 to 10 years of experience are most in demand. While an undergraduate degree provides an academic background to perform most responsibilities, for six of the nine core responsibilities an advanced degree was preferred by at least one-third of respondents. PMID- 10790790 TI - Listen for the "whispering disease". PMID- 10790789 TI - Health communication and consumer behavior on meat in Belgium: from BSE until dioxin. AB - This article focuses on the impact of mass media meat-health information on consumer perception, attitude, and behavior toward fresh meat in Belgium. In a situation similar to that which occurred in most other European countries, Belgian fresh meat consumption fell considerably during 1995-1999. A multitude of messages linking meat consumption to human health risks were reported by mass media. Bovine Spongiform Encephalopathy (BSE) since 1996 and dioxin in 1999 constituted the major issues. Empirical research, conducted in April 1998, revealed the tremendous negative impact of mass media coverage of meat-health issues on consumer risk perception, health concern, and attitude and behavior toward fresh meat. Oppositely, personal communication through butchers had only a small effect on consumer decision-making in this era dominated by alarming meat health press. Implications are threefold. First, mass media should be aware of its social responsibilities, which include spreading reliable and correct information to the society. This is especially the case as human health risks are involved. Second, the meat industry urgently needs to reorient itself toward quality, safety, and transparency. Finally, future communication dealing with similar crises situations requires cooperation across the meat chain, government, and those who are responsible for public health promotion and communication. PMID- 10790791 TI - Keeping in touch. Learn how to examine your breasts, then do it every month for a lifetime. PMID- 10790793 TI - Pain management in the addicted patient. PMID- 10790792 TI - Do I have breast cancer? PMID- 10790794 TI - Stop cervical cancer in its tracks. PMID- 10790795 TI - Monitoring albumin levels in phenytoin therapy. PMID- 10790796 TI - Nutrition matters. PMID- 10790797 TI - Evaluating neck masses. AB - Neck masses often present a diagnostic challenge to the primary care provider: Etiologies range from benign inflammations to life-threatening malignancies. Categorizing the etiologies into three broad categories is helpful when considering the extensive differential diagnosis for a neck mass: congenital, inflammatory/infectious, and neoplastic causes. This article discusses the evaluation and subsequent determination of whether a neck mass is significant and warrants further evaluation or is insignificant and may simply be observed. By performing a through history and physical examination, primary care providers can narrow the possibilities, differentiate between significant and insignificant neck masses, and select the appropriate treatment. PMID- 10790798 TI - Managing atopic dermatitis in children and adults. AB - Although the etiology of atopic dermatitis is not well understood, it appears to be linked to a combination of genetic and environmental factors, and it is usually associated with other atopic diseases such as asthma and hay fever. A definitive diagnosis in children and adults depends on identifying the nature and distribution of the lesions and on eliciting a personal or family history of the disease. Although no cure exists, atopic dermatitis often resolves spontaneously and can be controlled through proper management. Avoiding factors that precipitate or exacerbate inflammation is key to preventing disease flares. In children and adults, hydration and topical corticosteroids are the mainstays of therapy. Current advances in understanding the immunologic basis of the disease have led to the development of highly effective new treatments. Using patient education and support, the clinician can help adults and children successfully manage their disease. PMID- 10790799 TI - A primary care approach to cutaneous T-cell lymphoma. AB - Approximately 1,000 new cases of cutaneous T-cell lymphoma are definitively diagnosed each year. Mycosis fungoides and Sezary syndrome are the primary lymphomas in this group. Mycosis fungoides can begin in the patch, plaque, or tumor stage or in a combination. Less commonly, its initial presentation is erythrodermic. Because the initial appearance of cutaneous T-cell lymphoma can be subtle and the histopathologic evidence nonspecific, the disease is commonly misdiagnosed as a common dermatologic condition such as chronic eczema. Misdiagnosis can severely affect treatment and prognosis. Clinicians must be able to recognize this disease and know when to include it in the differential diagnosis. This article provides an overview of cutaneous T-cell lymphoma, discusses differential diagnoses, and outlines management considerations. PMID- 10790800 TI - Acute pharyngitis with lifelong implications. AB - A 32-year-old man presents with exudative pharyngitis and constitutional symptoms. This article discusses diagnostic pitfalls and explores the intricacies of therapeutic options. PMID- 10790801 TI - The evolving role of the acute care nurse practitioner. PMID- 10790802 TI - Homocysteine: a risk factor of atherosclerosis. PMID- 10790803 TI - Implementing the 1998 Canadian asthma guidelines. PMID- 10790804 TI - Interdisciplinary collaboration and primary health care reform Statement from the Ontario Chairs of Family Medicine and the Council of Ontario University Programs in Nursing. PMID- 10790805 TI - Travelers: know your medical history. PMID- 10790807 TI - Practising preventive medicine. PMID- 10790806 TI - Medical history in New Brunswick. PMID- 10790808 TI - Drug information incorrect. PMID- 10790809 TI - The College and the groves of Academe. PMID- 10790810 TI - Caffeine during pregnancy? In moderation. AB - QUESTION: Many of my female patients, those who plan pregnancy or have conceived, are afraid of any intake of caffeine. This often makes their lives miserable during pregnancy. Is this justified scientifically? ANSWER: Motherisk's recent meta-analysis suggests that the risks for miscarriage and fetal growth retardation increase only with daily doses of caffeine above 150 mg/d, equivalent to six typical cups of coffee a day. It is possible that some of this presumed risk is due to confounders, such as cigarette smoking. PMID- 10790811 TI - Ophthaproblem. Eales' disease. PMID- 10790812 TI - Emergency case. Renal colic. PMID- 10790813 TI - Practice tips. Deep venous thrombosis: before and after. Improving diagnosis and adjusting duration of therapy. PMID- 10790814 TI - Don't keep that ACE (inhibitor) up your sleeve! Is ramipril effective for secondary prevention of cardiovascular disease and stroke? PMID- 10790815 TI - Tretinoin and cutaneous photoaging. Guaranteed adverse effects! AB - * A cream containing 0.05% tretinoin (Retinova((R)) is approved for treatment of sun-induced skin damage ("photoaging").* Three trials comparing tretinoin with the excipient show that the effects of tretinoin cream are at best limited and slow to occur. Furthermore, they disappear on treatment cessation, necessitating long-term use.* The 0.05% tretinoin cream has poor local tolerability: most subjects develop irritation and fragile skin and require longer intervals between each application. Systemic adverse effects occur in some circumstances.* There are persistent doubts about whether it is safe to use tretinoin during pregnancy. PMID- 10790816 TI - Continuing educational needs in computers and informatics. McGill survey of family physicians. AB - OBJECTIVE: To describe family physicians' perceived educational needs in computers and informatics. DESIGN: Mailed survey. SETTING: General or family practices in Canada. PARTICIPANTS: Physicians (489 responded to a mailing sent to 2,500 physicians) who might attend sessions at the McGill Centre for CME. Two duplicate questionnaires were excluded from the analysis. METHOD: Four domains were addressed: practice profile, clinical CME needs, professional CME needs, and preferred learning formats. Data were entered on dBASE IV; analyses were performed on SPSS. MAIN FINDINGS: In the 487 questionnaires retained for analysis, "informatics and computers" was mentioned more than any other clinical diagnostic area, any other professional area, and all but three patient groups and service areas as a topic where improvement in knowledge and skills was needed in the coming year. Most physicians had no access to computer support for practice (62.6%); physicians caring for neonates, toddlers, or hospital inpatients were more likely to report some type of computer support. CONCLUSIONS: Family physicians selected knowledge and skills for computers and informatics as an area for improvement in the coming year more frequently than they selected most traditional clinical CME topics. This educational need is particularly great in small towns and in settings where some computerized hospital data are already available. PMID- 10790817 TI - Pilot study for appropriate anti-infective community therapy. Effect of a guideline-based strategy to optimize use of antibiotics. AB - OBJECTIVE: To determine whether a community-wide, multi-intervention educational strategy (CoMPLI model) could enhance adoption of clinical guidelines and improve the use of antibiotics. DESIGN: Before-after trial using baseline and study periods with a control group. SETTING: A small community in central Ontario. PARTICIPANTS: Health professionals, the general public, and the pharmaceutical industry. INTERVENTIONS: The educational strategy (CoMPLI), carried out during 6 winter months, consisted of continuing medical education sessions for health professionals and pharmaceutical representatives and a parallel public education campaign that included town hall meetings and pamphlets distributed by local pharmacists. The two main messages were: do not use antibiotics for viral respiratory infections, and use drugs recommended in the publication, Anti infective Guidelines for Community-Acquired Infections. MAIN OUTCOME MEASURES: Total number of antibiotic claims and adjusted odds ratios (OR) were used to measure the likelihood of physicians prescribing first- or second-line agents compared with the previous year and compared with control physicians. RESULTS: Claims in the study community decreased by nearly 10% during the 6-month study period compared with the baseline period from the previous year. Study physicians were 29% less likely (OR-1 = 0.71, range 0.67 to 0.76) to prescribe second-line antibiotics during the study period than physicians in the rest of the province. CONCLUSIONS: Physicians participating in the pilot study were more likely to follow drug recommendations outlined in published guidelines. PMID- 10790818 TI - Seeking medical advice if HIV symptoms are suspected. Qualitative study of beliefs among HIV-negative gay men. AB - OBJECTIVE: To identify beliefs associated with seeking medical advice promptly when symptoms of HIV infection are suspected among HIV-negative gay men. DESIGN: Qualitative study of beliefs among focus group participants. SETTING: Quebec city, Que, metropolitan area. PARTICIPANTS: Referred sample of 20 HIV-negative gay men 18 to 45 years old who attended bars, university, or gay associations in Quebec city. METHOD: Three focus groups of five to seven subjects were formed and each 2-hour session was tape-recorded. MAIN OUTCOME FINDINGS: Participants thought that seeking medical advice promptly when symptoms of HIV are suspected would help them be informed about their health status and would eliminate unnecessary anxiety and fear, but would force them to face reality and make major changes. Barriers were the quality of the relationship with their physicians and concern about discussing their sexual lives. Normative beliefs were sought from members of community groups, circles of close friends, health providers, and the media. CONCLUSIONS: Several beliefs could influence the motivation of seronegative gay men to seek medical advice promptly when symptoms of HIV infection are suspected. These beliefs should be integrated into programs promoting early consultation with physicians and into clinical counseling, as integration could facilitate early treatment and care. Physicians should give special attention to establishing relationships of trust with these patients. PMID- 10790820 TI - Case report: allergic bronchopulmonary aspergillosis in asthma. PMID- 10790819 TI - Leukotriene-receptor antagonists. Role in asthma management. AB - OBJECTIVE: To examine the role of leukotriene-receptor antagonists (LTRAs) in management of asthma. QUALITY OF EVIDENCE: Most data were derived from randomized, double-blind, controlled trials. MAIN MESSAGE: Leukotrienes appear to have an important role in the pathophysiology of asthma, including airway inflammation. Leukotriene-receptor antagonists are effective in improving asthma control end points, such as allergen, ASA, and exercise challenge, in clinical models of asthma. In chronic asthma, LTRA administration reduces asthma symptoms and rescue beta 2-agonist use, changes that are paralleled by improvements in lung function. Both zafirlukast and montelukast decrease circulating levels of eosinophils and could have other useful anti-inflammatory properties. Administration of LTRAs allows doses of inhaled corticosteroids to be reduced. Currently available LTRAs are free of serious side effects and are available as oral formulations. CONCLUSIONS: Leukotriene-receptor antagonists belong to a new class of asthma medication. While inhaled corticosteroids remain first-line therapy for managing chronic asthma, LTRAs should be considered for patients with ASA-sensitive asthma; as adjunct therapy when low to moderate doses of inhaled steroid alone provide incomplete control; or as adjunct therapy to allow reduction in doses of inhaled corticosteroids. PMID- 10790821 TI - Endometrial cancer. Prevention, detection, management, and follow up. AB - OBJECTIVE: To review risk factors for uterine cancer; to discuss strategies for detecting uterine cancer; to outline prognostic factors and treatment; and to review the role of follow up for patients who have completed primary therapy. QUALITY OF EVIDENCE: MEDLINE was searched from January 1996 to June 1998 using the terms endometrial neoplasms, estrogen replacement therapy, hormone replacement therapy, tamoxifen, and screening. Only English language articles were reviewed. Study types included reviews. Bibliographies of articles found were searched for further relevant titles. Causation literature is available from well conducted cohort trials. Treatment recommendations are based in part on prognostic information and a few randomized controlled trials. MAIN MESSAGE: Risk factors, both intrinsic and extrinsic, are associated with uterine cancer. Family physicians have a role in preventing disease by ensuring that all women with uteri in situ using hormone replacement therapy (HRT) have progesterone therapy as part of the HRT regimen. Detection is crucial; abnormal uterine bleeding or undiagnosed postmenopausal bleeding warrants investigation with endometrial biopsy. The goal of surgery is to remove the uterus and ovaries and identify factors that make the disease at high risk of recurrence. Although adjuvant radiation therapy does not prolong survival, it does alter the pattern of disease recurrence. The goal of follow up after primary therapy is to identify recurrent disease while it is still curable. CONCLUSIONS: Family physicians play an important role in preventing uterine cancer, initiating early diagnosis of disease, and in the future, might be more actively involved in caring for patients following primary therapy. PMID- 10790823 TI - Estimation of in situ elastic properties of biphasic cartilage based on a transversely isotropic hypo-elastic model. AB - Articular cartilage is known to behave nonlinearly for large deformations. Mechanical properties derived from small strain experiments yield excessively large deformations in finite element models used in the study of severe blunt impact to joints. In this manuscript, a method is presented to determine the nonlinear elastic properties of biphasic cartilage based on a transversely isotropic hypo-elastic model. The elastic properties were estimated by fitting two force-displacement curves (in rapid loading and at equilibrium) obtained from large deformation indentation relaxation tests on cartilage using a nonporous spherical indentor. The solid skeleton of the cartilage was modeled as a transversely isotropic hypo-elastic material and a commercial finite element program was employed to solve the problem of a layer indented by a rigid sphere. Components of the hypo-elasticity tensor were made dependent on deformation according to the variations defined by a transversely isotropic hyperelastic formulation given earlier by others. Material incompressibility was assumed during the initial stage of rapid loading. The analysis was utilized for the determination of in situ properties of rabbit retropatellar cartilage at large deformations. The model was able to fit the material response to rapid loading and equilibrium indentation test data to approximately 50 percent strain. This material model suggested even higher percentage of stress supported by the fluid phase of cartilage than given earlier by small deformation theories of biphasic cartilage. PMID- 10790822 TI - Controlling asthma. Highlights of the 1999 Canadian Asthma Consensus Report. PMID- 10790824 TI - Quantifying skeletal muscle properties in cadaveric test specimens: effects of mechanical loading, postmortem time, and freezer storage. AB - Investigators currently lack the data necessary to define the state of skeletal muscle properties within cadaveric specimens. The purpose of this study is to define the temporal changes in the postmortem properties of skeletal muscle as a function of mechanical loading and freezer storage. The tibialis anterior of the New Zealand white rabbit was chosen for study. Modulus and no-load strain were found to vary significantly from live after eight hours postmortem. Following the changes that occur during rigor mortis, a stable region of postmortem, post-rigor properties occurred between 36 to 72 hours postmortem. A freeze-thaw process was not found to have a significant effect on the post-rigor response. The first loading cycle response of post-rigor muscle was unrepeatable but stiffer than live passive muscle. After preconditioning, the post-rigor muscle response was repeatable. The preconditioned post-rigor response was less stiff than the live passive response due to a significant increase in no-load strain. Failure properties of postmortem muscle were found to be significantly different from live passive muscle with a significant decrease in failure stress (61 percent) and energy (81 percent), while failure strain was unchanged. These results suggest that the post-rigor response of cadaveric muscle is unaffected by freezing but sensitive to even a few cycles of mechanical loading. PMID- 10790825 TI - Linear and quasi-linear viscoelastic characterization of ankle ligaments. AB - The objective of this study was to produce linear and nonlinear viscoelastic models of eight major ligaments in the human ankle/foot complex for use in computer models of the lower extremity. The ligaments included in this study were the anterior talofibular (ATaF), anterior tibiofibular (ATiF), anterior tibiotalar (ATT), calcaneofibular (CF), posterior talofibular (PTaF), posterior tibiofibular (PTiF), posterior tibiotalar (PTT), and tibiocalcaneal (TiC) ligaments. Step relaxation and ramp tests were performed. Back-extrapolation was used to correct for vibration effects and the error introduced by the finite rise time in step relaxation tests. Ligament behavior was found to be nonlinear viscoelastic, but could be adequately modeled up to 15 percent strain using Fung's quasilinear viscoelastic (QLV) model. Failure properties and the effects of preconditioning were also examined. PMID- 10790826 TI - Biaxial mechanical properties of the natural and glutaraldehyde treated aortic valve cusp--Part I: Experimental results. AB - To date, there are no constitutive models for either the natural or bioprosthetic aortic valve (AV), in part due to experimental complications related to the AV's small size and heterogeneous fibrous structure. In this study, we developed specialized biaxial testing techniques for the AV cusp, including a method to determine the local structure-strain relationship to assess the effects of boundary tethering forces. Natural and glutaraldehyde (GL) treated cusps were subjected to an extensive biaxial testing protocol in which the ratios of the axial tensions were held at constant values. Results indicated that the local fiber architecture clearly dominated cuspal deformation, and that the tethering effects at the specimen boundaries were negligible. Due to unique aspects of cuspal fiber architecture, the most uniform region of deformation was found at the lower portion as opposed to the center of the cuspal specimen. In general, the circumferential strains were much smaller than the radial strains, indicating a profound degree of mechanical anisotropy, and that natural cusps were significantly more extensible than the GL treated cusps. Strong mechanical coupling between biaxial stretch axes produced negative circumferential strains under equibiaxial tension. Further, the large radial strains observed could not be explained by uncrimping of the collagen fibers, but may be due to large rotations of the highly aligned, circumferential-oriented collagen fibers in the fibrosa. In conclusion, this study provides new insights into the AV cusp's structure-function relationship in addition to requisite data for constitutive modeling. PMID- 10790828 TI - A quantitative model of cellular elasticity based on tensegrity. AB - A tensegrity structure composed of six struts interconnected with 24 elastic cables is used as a quantitative model of the steady-state elastic response of cells, with the struts and cables representing microtubules and actin filaments, respectively. The model is stretched uniaxially and the Young's modulus (E0) is obtained from the initial slope of the stress versus strain curve of an equivalent continuum. It is found that E0 is directly proportional to the pre existing tension in the cables (or compression in the struts) and inversely proportional to the cable (or strut) length square. This relationship is used to predict the upper and lower bounds of E0 of cells, assuming that the cable tension equals the yield force of actin (approximately 400 pN) for the upper bound, and that the strut compression equals the critical buckling force of microtubules for the lower bound. The cable (or strut) length is determined from the assumption that model dimensions match the diameter of probes used in standard mechanical tests on cells. Predicted values are compared to reported data for the Young's modulus of various cells. If the probe diameter is greater than or equal to 3 microns, these data are closer to the lower bound than to the upper bound. This, in turn, suggests that microtubules of the CSK carry initial compression that exceeds their critical buckling force (order of 10(0)-10(1) pN), but is much smaller than the yield force of actin. If the probe diameter is less than or equal to 2 microns, experimental data fall outside the region defined by the upper and lower bounds. PMID- 10790827 TI - Prevention of mechanical stretch-induced endothelial and smooth muscle cell injury in experimental vein grafts. AB - Vein grafts are subject to increased tensile stress due to exposure to arterial blood pressure, which has been hypothesized to induce endothelial cell (EC) and smooth muscle cell (SMC) injury. This study was designed to verify this hypothesis and to develop a tissue engineering approach that can be used to prevent these pathological events. Two experimental models were created in rats to achieve these goals: (1) a nonengineered vein graft with increased tensile stress, which was created by grafting a jugular vein into the abdominal aorta using a conventional end-to-end anastomotic technique; and (2) an engineered vein graft with reduced tensile stress, which was created by restricting a vein graft into a cylindrical sheath constructed using a polytetrafluoroethylene membrane. The integrity of ECs in these models was examined by using a silver nitrate staining method, and the integrity of SMCs was assessed by using a fluorescein phalloidin-labeling technique. It was found that nonengineered vein grafts were associated with early EC denudation with a change in EC coverage from 100 percent in normal jugular veins to 36 +/- 10, 28 +/- 12, 18 +/- 9, 44 +/- 15, 80 +/- 13, and 97 +/- 6 percent at 1 and 6 hours and 1, 5, 10, and 30 days, respectively. Similarly, rapid SMC actin filament degradation was found during the early period with a change in SMC coverage from approximately 94 percent in normal jugular veins to 80 +/- 10, 41 +/- 17, 25 +/- 9, 51 +/- 15, 79 +/- 15, 98 +/- 2 percent at 1 and 6 hours and 1, 5, 10, and 30 days, respectively, in nonengineered vein grafts. In engineered vein grafts with reduced tensile stress, EC denudation and SMC actin filament degradation were prevented significantly. These results suggested that mechanical stretch due to increased tensile stress contributed to EC and SMC injury in experimental vein grafts, and these pathological events could be partially prevented when tensile stress was reduced by using a biomechanical engineering approach. PMID- 10790829 TI - A finite element method for mechanical response of hair cell ciliary bundles. AB - This paper describes the development of a methodology for performing a mechanical analysis of hair cell ciliary bundles. The cilia were modeled as shear deformable beams, and interconnections were modeled as two-force members. These models were incorporated into software, which performs a finite element analysis of a user defined bundle. The algorithm incorporates aspects of the bundle such as geometric realignment and buckling of compressed side links. A sample bundle is introduced and results of modeling it are presented. PMID- 10790830 TI - Supraphysiological thermal injury in Dunning AT-1 prostate tumor cells. AB - To investigate the potential application of thermal therapy in the treatment of prostate cancer, the effects of supraphysiological temperatures (40-70 degrees C) for clinically relevant time periods (approximately 15 minutes) were experimentally studied on attached Dunning AT-1 rat prostate cancer cells using multiple assays. The membrane and reproductive machinery were the targets of injury selected for this study. In order to assess membrane injury, the leakage of calcein was measured dynamically, and the uptake of PI was measured postheating (1-3 hours). Clonogenicity was used as a measure of injury to the reproductive machinery 7 days post-injury after comparable thermal insults. Experimental results from all three assays show a broad trend of increasing injury with an increase in temperature and time of insult. Membrane injury, as measured by the fluorescent dye assays, does not correlate with clonogenic survival for many of the thermal histories investigated. In particular, the calcein assay at temperatures of < or = 40 degrees C led to measurable injury accumulation (dye leakage), which was considered sublethal, as shown by significant survival for comparable insult in the clonogenic assay. Additionally, the PI uptake assay used to measure injury post-thermal insult shows that membrane injury continues to accumulate after thermal insult at temperatures > or = 50 degrees C and may not always correlate with clonogenicity at hyperthermic temperatures such as 45 degrees C. Last, although the clonogenic assay yields the most accurate cell survival data, it is difficult to acquire these data at temperatures > or = 50 degrees C because the thermal transients in the experimental setup are significant as compared to the time scale of the experiment. To improve prediction and understanding of thermal injury in this prostate cancer cell line, a first-order rate process model of injury accumulation (the Arrhenius model) was fit to the experimental results. The activation energy (E) obtained using the Arrhenius model for an injury criterion of 30 percent for all three assays revealed that the mechanism of thermal injury measured is likely different for each of the three assays: clonogenics (526.39 kJ/mole), PI (244.8 kJ/mole), and calcein (81.33 kJ/mole). Moreover, the sensitivity of the rate of injury accumulation (d omega/dt) to temperature was highest for the clonogenic assay, lowest for calcein leakage, and intermediate for PI uptake, indicating the strong influence of E value on d omega/dt. Since the clonogenic assay is linked to the ultimate survival of the cell and accounts for all lethal mechanisms of cellular injury, the E and A values obtained from clonogenic study are the best values to apply to predict thermal injury in cells. For higher temperatures (> or = 50 degrees C) indicative of thermal therapies, the results of PI uptake can be used as a conservative estimate of cell death (underprediction). This is useful until better experimental protocols are available to account for thermal transients at high temperature to assess clonogenic ability. These results provide further insights into the mechanisms of thermal injury in single cell systems and may be useful for designing optimal protocols for clinical thermal therapy. PMID- 10790831 TI - Biomechanical study of pediatric human cervical spine: a finite element approach. AB - Although considerable effort has been made to understand the biomechanical behavior of the adult cervical spine, relatively little information is available on the response of the pediatric cervical spine to external forces. Since significant anatomical differences exist between the adult and pediatric cervical spines, distinct biomechanical responses are expected. The present study quantified the biomechanical responses of human pediatric spines by incorporating their unique developmental anatomical features. One-, three-, and six-year-old cervical spines were simulated using the finite element modeling technique, and their responses computed and compared with the adult spine response. The effects of pure overall structural scaling of the adult spine, local component developmental anatomy variations that occur to the actual pediatric spines, and structural scaling combined with local component anatomy variations on the responses of the pediatric spines were studied. Age- and component-related developmental anatomical features included variations in the ossification centers, cartilages, growth plates, vertebral centrum, facet joints, and annular fibers and nucleus pulposus of the intervertebral discs. The flexibility responses of the models were determined under pure compression, pure flexion, pure extension, and varying degrees of combined compression-flexion and compression-extension. The pediatric spine responses obtained with the pure overall (only geometric) scaling of the adult spine indicated that the flexibilities consistently increase in a uniform manner from six- to one-year-old spines under all loading cases. In contrast, incorporation of local anatomic changes specific to the pediatric spines of the three age groups (maintaining the same adult size) not only resulted in considerable increases in flexibilities, but the responses also varied as a function of the age of the pediatric spine and type of external loading. When the geometric scaling effects were added to these spines, the increases in flexibilities were slightly higher; however, the pattern of the responses remained the same as found in the previous approach. These results indicate that inclusion of developmental anatomical changes characteristic of the pediatric spines has more of a predominant effect on biomechanical responses than extrapolating responses of the adult spine based on pure overall geometric scaling. PMID- 10790832 TI - A reconciliation of local and global models for bone remodeling through optimization theory. AB - Remodeling rules with either a global or a local mathematical form have been proposed for load-bearing bones in the literature. In the local models, the bone architecture (shape, density) is related to the strains/energies sensed at any point in the bone, while in the global models, a criterion believed to be applicable to the whole bone is used. In the present paper, a local remodeling rule with a strain "error" form is derived as the necessary condition for the optimum of a global remodeling criterion, suggesting that many of the local error driven remodeling rules may have corresponding global optimization-based criteria. The global criterion proposed in the present study is a trade-off between the cost of metabolic growth and use, mathematically represented by the mass, and the cost of failure, mathematically represented by the total strain energy. The proposed global criterion is shown to be related to the optimality criteria methods of structural optimization by the equivalence of the model solution and the fully stressed solution for statically determinate structures. In related work, the global criterion is applied to simulate the strength recovery in bones with screw holes left behind after removal of fracture fixation plates. The results predicted by the model are shown to be in good agreement with experimental results, leading to the conclusion that load-bearing bones are structures with optimal shape and property for their function. PMID- 10790833 TI - Numerical simulation of pulsatile flow in a compliant curved tube model of a coronary artery. AB - The endothelial cells (ECs) lining a blood vessel wall are exposed to both the wall shear stress (WSS) of blood flow and the circumferential strain (CS) of pulsing artery wall motion. These two forces and their interaction are believed to play a role in determining remodeling of the vessel wall and development of arterial disease (atherosclerosis). This study focused on the WSS and CS dynamic behavior in a compliant model of a coronary artery taking into account the curvature of the bending artery and physiological radial wall motion. A three dimensional finite element model with transient flow and moving boundaries was set up to simulate pulsatile flow with physiological pressure and flow wave forms characteristic of the coronary arteries. The characteristic coronary artery curvature and flow conditions applied to the simulation were: aspect ratio (lambda) = 10, diameter variation (DV) = 6 percent, mean Reynolds number (Re) = 150, and unsteadiness parameter (alpha) = 3. The results show that mean WSS is about 50 percent lower on the inside wall than the outside wall while WSS oscillation is stronger on the inside wall. The stress phase angle (SPA) between CS and WSS, which characterizes the dynamics of the mechanical force pattern applied to the endothelial cell layer, shows that CS and WSS are more out of phase in the coronaries than in any other region of the circulation (-220 deg on the outside wall, -250 deg on the inside wall). This suggests that in addition to WSS, SPA may play a role in localization of coronary atherosclerosis. PMID- 10790834 TI - The influence of out-of-plane geometry on the flow within a distal end-to-side anastomosis. AB - This paper describes a computational and experimental investigation of flow in a proto-type model geometry of a fully occluded 45 deg distal end-to-side anastomosis. Previous investigations have considered a similar configuration where the centerlines of the bypass and host vessels lie within a plane, thereby producing a plane of symmetry within the flow. We have extended these investigations by deforming the bypass vessel out of the plane of symmetry, thereby breaking the symmetry of the flow and producing a nonplanar geometry. Experimental data were obtained using magnetic resonance imaging of flow within perspex models and computational data were obtained from simulations using a high order spectral/hp element method. We found that the nonplanar three-dimensional flow notably alters the distribution of wall shear stress at the bed of the anastomosis, reducing the peak wall shear stress peak by approximately 10 percent when compared with the planar model. Furthermore, an increase in the absolute flux of velocity into the occluded region, proximal to the anastomosis, of 80 percent was observed in the nonplanar geometry when compared with the planar geometry. PMID- 10790835 TI - Rapid three-dimensional segmentation of the carotid bifurcation from serial MR images. AB - The current trend in computational hemodynamics is to employ realistic models derived from ex vivo or in vivo imaging. Such studies typically produce a series of images from which the lumen boundaries must first be individually extracted (i.e., two-dimensional segmentation), and then serially reconstructed to produce the three-dimensional lumen surface geometry. In this paper, we present a rapid three-dimensional segmentation technique that combines these two steps, based on the idea of an expanding virtual balloon. This three-dimensional technique is demonstrated in application to finite element meshing and CFD modeling of flow in the carotid bifurcation of a normal volunteer imaged with black blood MRI. Wall shear stress patterns computed using a mesh generated with the three-dimensional technique agree well with those computed using a mesh generated from conventional two-dimensional segmentation and serial reconstruction. In addition to reducing the time required to extract the lumen surface from hours to minutes, our approach is easy to learn and use and requires minimal user intervention, which can potentially increase the accuracy and precision of quantitative and longitudinal studies of hemodynamics and vascular disease. PMID- 10790836 TI - Microcirculation impedance analysis in cat lung. AB - An analysis of pulsatile microcirculation in cat lung, with special attention to the pulmonary microvascular impedance, is presented. A theoretical calculation is made on the basis of a complete set of experimental data on the morphology and elasticity of cat's pulmonary capillary sheets. The transfer matrix of the pulmonary microvascular impedance is obtained. The input impedance at the capillary entrance and exit are determined. The input impedance at the pulmonary arterial trunk is compared under various physiological conditions. It is shown that although the impact of pulmonary microcirculation on the relationship between the steady mean flow and pressure in the pulmonary arteries and veins is decisively large, the influence of the alveolar microcirculation on the input impedance at the pulmonary arterial trunk is small. PMID- 10790837 TI - The role of process evaluation in the training of facilitators for an adolescent health education program. AB - This article reports on the process evaluation of the training of facilitators for the Adolescent Social Action Program, a health education program in Albuquerque, New Mexico that trained college students and adult volunteers to work with middle school students. From the process evaluation data collected throughout a four-year period (1995-1998), data relevant to training are described: facilitator characteristics, facilitator training, curriculum implementation, and use of the program's model designed to promote critical thinking and dialogue. Results indicated that, though most facilitators reported the training was sufficient to enable them to implement the curriculum, they did not completely do so, especially as groups reached their final sessions. Facilitators covered the core curriculum content, but often failed to follow through with the more abstract activities. The need to perform and report the process evaluation in time to provide ample opportunity for trainers and curriculum designers to make appropriate adjustments is discussed. PMID- 10790838 TI - The SiteLegend: twelve components of a new strategy for providing website documentation. AB - The World Wide Web continues to evolve dramatically with an estimated 57 million users in the United States. Some two-thirds of Internet users in the United States searched for health information the past year, with 90% indicating they found the information they sought. Thus, health and medical information represents the single most popular topic on the Web, surpassing all other categories. Yet, few measures exist to ensure the accuracy of information provided to children and adults through such sites. Some existing quality control measures such as assessment instruments, authoritative guidelines, and seal of approval systems provide screening mechanisms, but none of these measures guarantees protection for users. This article proposes a new approach to ensuring full and accurate disclosure of information about a website. Referred to as a SiteLegend, the approach uses a hyperlink to fully inform consumers regarding 12 essential components of website. This article describes each of the 12 essential components, then provides an example of an active online link that illustrates actual use of the SiteLegend hyperlink approach. PMID- 10790839 TI - An in-school CD-ROM asthma education program. AB - Computer-based education in schools is not novel. However, only a few computer games have been introduced into school-based health education programs. This study describes inclusion of an asthma education space adventure game into fourth grade classrooms. Using the game improved asthma knowledge in the intervention classes. Children in the active participation classroom gained significantly more asthma knowledge over the observation period compared to the classroom that did not play the computer game. Knowledge gained was retained over a four-week period and the addition of physician-led talks appeared to add little to the knowledge gained. "Air Academy: The Quest for Airtopia" is an easy and successful tool for elementary grade asthma-related health education in schools. PMID- 10790840 TI - Entropy measurements and the kinetic method: a statistically meaningful approach AB - In the literature, data obtained using the kinetic method have been analyzed to provide both ion affinities and relative entropies for the competitive dissociations involved. In this work, the procedure used to extract this information is shown to be statistically flawed. Using more rigorous statistical procedures, we outline alternative methods of acquiring the same information, including straightforward means of analyzing the uncertainties in the thermodynamic quantities obtained. Fortunately, it is expected that the central values reported in previous work need not be changed, but the uncertainties are much larger than has been previously detailed. The validity of the assumptions involved in the extraction of entropy effects is discussed in some detail. PMID- 10790841 TI - Protonation sites in methyl nitrate and the formation of transient CH4NO3 radicals. A neutralization-reionization mass spectrometric and computational study AB - Protonation sites in methyl nitrate (1) were evaluated computationally at the Gaussian 2(MP2) level of ab initio theory. The methoxy oxygen was the most basic site that had a calculated proton affinity of PA = 728-738 kJ mol-1 depending on the optimization method used to calculate the equilibrium geometry of the CH3O(H) NO2+ ion (2+). Protonation at the terminal oxygen atoms in methyl nitrate was less exothermic; the calculated proton affinities were 725, 722, and 712 kJ mol-1 for the formation of the syn-syn, anti-syn, and syn-anti ion rotamers 3a+, 3b+, and 3c+, respectively. Ion 2+ was prepared by an ion-molecule reaction of NO2+ with methanol and used to generate the transient CH3O(H)-NO2. radical (2) by femtosecond collisional electron transfer. Exothermic protonation of 1 produced a mixture of 3a(+)-3c+ with 2+ that was used to generate transient radicals 3a-3c. Radical 2 was found to be unbound and dissociated without barrier to methanol and NO2. Radicals 3a-3c were calculated to be weakly bound. When formed by vertical neutralization, 3a-3c dissociated completely on the 4.2 microseconds time scale of the experiment. The main dissociations of 3a-3c were formations of CH3O. + HONO and CH3ONO + OH.. The gas-phase chemistry of radicals 3a-3c and their dissociation products, as studied by neutralization-reionization mass spectrometry, was dominated by Franck-Condon effects on collisional neutralization and reionization. The adiabatic ionization energies of 3a-3c were calculated as 7.54, 7.57, and 7.66 eV, respectively. PMID- 10790842 TI - Single isolated droplets with net charge as a source of ions AB - A levitation device for charged particles has been used to position a single isolated droplet at a time at atmospheric pressure near the sampling orifice of a vacuum chamber. Following a brief desolvation period (550 ms), a series of coluomb fission events were initiated. Several atmospheric pressure electrode designs were evaluated with respect to guiding the progeny droplets/ions, to the sampling orifice. The best design tested consisted of a series of four annular electrodes of decreasing radius positioned above the levitation ring electrodes, and, on average, 40 ions were counted per single isolated droplet. The ion utilization (charge detected versus charge in the original droplet) with this electrode design has been estimated to be 5 x 10(-6), a substantial improvement over the current utilization measured to be < or = 1 x 10(-9) with a conventional electrospray ion source using the same vacuum apparatus. PMID- 10790843 TI - An innovative method for measuring hydrogen and deuterium: chemical reaction interface mass spectrometry with nitrogen reactant gas. AB - A new method for measuring deuterium isotopic enrichment with CRIMS (chemical reaction interface mass spectrometry) is described. Using nitrogen as the reactant gas in a chemical reaction interface generates molecular hydrogen that provides the H2 and HD from which the deuterium content can be analyzed with a benchtop quadrupole mass spectrometer. Samples of deuterated leucine in unlabeled leucine were used as the primary test species. Detection of deuterium enrichment was accurate, precise, and linear. We used this scheme to evaluate the results of a process to acetylate lysine residues in a peptide-neurotensin. With separation on a C18 column, we found a 61% yield of the desired monoethylated product that had a D/H ratio very close to the theoretical one. Isotope ratio monitoring for deuterated species will be important in metabolism studies where CRIMS generates a comprehensive and quantitative view of products of deuterated precursors. Where concerns about metabolic isotope effects of deuterium are absent, the use of deuterium will enable these studies to be performed with simpler syntheses and at less cost than if using 13C or 15N. PMID- 10790844 TI - Mass resolution of 11,000 to 22,000 with a multiple pass quadrupole mass analyzer AB - CO+ and N+2 are separated with resolution of 11,000 [full width half maximum (FWHM)] using a conventional quadrupole mass spectrometer by applying square wave voltages to the entrance and exit lenses to trap or reflect the ions for multiple passes. A resolution of 22,000 (FWHM) with 63% of the total signal remaining is attained using multiple passes when ions are stored between injection pulses. Gated ion extraction also reduces the mass shift and number and intensity of artifact peaks and permits better resolution compared to the performance obtained when the ions are injected continuously. PMID- 10790845 TI - Obtaining more accurate Fourier transform ion cyclotron resonance mass measurements without internal standards using multiply charged ions. AB - Space-charge effects produce frequency shifts in Fourier transform ion cyclotron resonance (FTICR) mass spectrometry and correction for these shifts is necessary for obtaining accurate mass measurements. We report a novel method for obtaining accurate mass calibration to correct for space-charge induced mass shifts without the requirement for internal calibrants. The new approach is particularly well suited for electrospray ionization-FTICR mass spectra that contain multiple charge states of the same molecular species. This method, deconvolution of Coulombic affected linearity (DeCAL), is described and presented with several examples demonstrating the increased mass measurement accuracy obtained. DeCAL provides the basis for more routinely obtaining higher mass accuracy measurements in conjunction with chromatographic separations for complex mixture analysis, and obviates the need for internal calibration in many applications. PMID- 10790846 TI - Method for screening peptide fragment ion mass spectra prior to database searching. AB - A methodology is described for screening fragment ion spectra of peptides prior to database searching for protein identification. A software routine written in the Perl programming language was used to analyze data from previous Sequest database searches and develop a set of statistical descriptors that could be used to identify spectra not likely to yield useful results in a database search. A second Perl program used an evolutionary algorithm to optimize the criteria for each statistical descriptor and generate a formula for determining spectral quality. This formula was used by a third Perl program to screen data sets from four independent liquid chromatography tandem mass spectrometry runs. On the average, use of the screening program reduced the time required for a database search by 1/2 with little loss of useful information from the database search results. PMID- 10790847 TI - Structure and fragmentation of b2 ions in peptide mass spectra. AB - In a number of cases the b2 ion observed in peptide mass spectra fragments directly to the a1 ion. The present study examines the scope of this reaction and provides evidence as to the structure(s) of the b2 ions undergoing fragmentation to the a1 ion. The b2 ion H-Ala-Gly+ fragments, in part, to the a1 ion, whereas the isomeric b2 ion H-Gly-Ala+ does not fragment to the a1 ion. Ab initio calculations of ion energies show that this different behavior can be rationalized in terms of protonated oxazolone structures for the b2 ions provided one assumes a reverse activation energy of approximately 1 eV for the reaction b2 ->a2; such a reverse activation energy is consistent with experimental kinetic energy release measurements. Experimentally, the H-Aib-Ala+ b2 ion, which must have a protonated oxazolone structure, fragments extensively to the a1 ion. We conclude that the proposal by Eckart et al. (J. Am. Soc. Mass Spectrom. 1998, 9, 1002) that the b2 ions which undergo fragmentation to a1 ions have an immonium ion structure is not necessary to rationalize the results, but that the fragmentation does occur from a protonated oxazolone structure. It is shown that the b2-->a1 reaction occurs extensively when the C-terminus residue in the b2 ion is Gly and with less facility when the C-terminus residue is Ala. When the C terminus residue is Val or larger, the b2-->a1 reaction cannot compete with the b2-->a2 fragmentation reaction. Some preliminary results on the fragmentation of a2 ions are reported. PMID- 10790848 TI - Structural determination of sphingomyelin by tandem mass spectrometry with electrospray ionization. AB - Alkaline metal adduct ions of sphingomyelin were formed by electrospray ionization in positive ion mode. Under low energy collisionally activated dissociation (CAD), the product ion spectra yield abundant fragment ions representative of both long chain base and fatty acid which permit unequivocal determination of the structure. Tandem spectra obtained by constant neutral loss scanning permit identification of sphingomyelin class and specific long chain base subclass in the mixture. The fragmentation pathways under CAD were proposed, and were further confirmed by source CAD tandem mass spectrometry. The total analysis of sphingomyelin mixtures from bovine brain, bovine erythrocytes, and chicken egg yolk is also presented. PMID- 10790849 TI - Gas-phase stability of double-stranded oligodeoxynucleotides and their noncovalent complexes with DNA-binding drugs as revealed by collisional activation in an ion trap. AB - The intrinsic (gas-phase) stabilities of duplex, self-complementary oligonucleotides were measured in a relative way by subjecting the duplex precursor ions to increasing amounts of collision energy during the collisional activated decomposition (CAD) events in an ion-trap mass spectrometer. The results are displayed as a dissociation profile, an s-shaped curve that shows the dependence of the relative abundance of the duplex on the applied collision energy. The total number of charges, the total number of base pairs, and the location of the high proton-affinity bases (i.e., G and C) are the main factors that affect the intrinsic stability of the duplex oligonucleotides. If the charge state is the same, the stability, as measured as a half-wave collision energy, E1/2, correlates well with the total number of H bonds for the duplex. The intrinsic stabilities of noncovalent complexes between duplex oligonucleotide and some DNA-binding drugs were also measured by using the newly developed method. Although duplexes are stabilized in the gas phase when they bind to drug molecules, correlations between gas-phase stabilities and the solution-binding affinities have not yet been obtained. Complexes in which the drug is bound in the minor groove must be joined tightly because they tend to dissociate in the gas phase by breaking covalent bonds of the oligonucleotide to give base loss and small sequence-ion formation. Complexes in which the drug is known to favor intercalation dissociate by breaking weak, noncovalent bonds to form single stranded oligonucleotides although cleavage of covalent bonds of the oligonucleotide also occurs. PMID- 10790850 TI - Accurate mass measurement of low molecular weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - We report the application of matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS) for the accurate measurement of mass of low molecular weight compounds (smaller than 1500 Da), a linear peptide, two types of cyclic depsipeptides, a polyhydroxy-macrocyclic lactone, and two prenylated flavonoids, with delayed extraction in the reflector mode. The performance of the MALDI-TOF instrument was less than those of fast atom bombardment and Fourier-transform ion cyclotron resonance mass spectrometry instruments and insufficient to give acceptable accuracy for literature reporting. Nevertheless, when combined with NMR spectrometry and/or amino acid analysis to give information on the numbers of carbon atoms and index of hydrogen deficiency, MALDI was useful for determination of the elemental composition of the low molecular weight compounds available in small quantities. PMID- 10790851 TI - Generation of electrospray from a solution predeposited on optical fibers coiled with a platinum wire. AB - This study examines the feasibility of generating electrospray directly from the tip of two optical fibers bound together with Teflon tape. This approach does not require a capillary and syringe pump. The electrospray source is simply constructed by coiling the two optical fibers with a platinum (Pt) wire. The optical fibers extend beyond the Pt coil for approximately 1 cm. The sample solution is predeposited on the Pt coil by a micropipette. As the high voltage required for electrospray is applied to the coil, the sample solution moves along the grooves between the two optical fibers. A stable electrospray is subsequently generated at the tip of the fibers. The mass spectra of insulin, lysozyme, and ubiquitin are exactly the same as those obtained by conventional electrospray using a capillary and syringe pump. Rapid determination of the active ingredient in a tablet by this technique is demonstrated. PMID- 10790852 TI - Hyperoxia-enhanced activation-induced hemodynamic response in human VI: an fMRI study. AB - The effect hyperoxia had on the hemodynamic response to visual stimulation (black and white checkerboard alternating at a frequency of 8 Hz) of human VI was investigated using a blood oxygenation level-dependent (BOLD) contrast with an fMRI technique. Data were acquired with a 5 on/5 off block paradigm using single shot gradient-echo echo-planar imaging. Using a two-tailed paired t-test (p < 0.05, n = 13) it was found that the mean percentage signal change and the mean number of activated pixels was significantly increased for hyperoxia (5.7 +/- 0.9, 187 +/- 73, mean +/- SD) relative to those for normoxia (5.4 +/- 0.9, 168 +/ 58). We believe that these results indicate that hyperoxia enhances the activation-induced hemodynamic response in human VI. PMID- 10790854 TI - Brain stem auditory evoked fields in response to clicks. AB - Magnetoencephalographic correlates of brain stem auditory evoked potentials (BAEP) have been identified by applying the model of a current dipole with invariant location and orientation (fixed dipole) to data recorded with a 37 channel first-order gradiometer system. In all three subjects studied, the dominating wave of the recorded brain stem auditory evoked field (BAEF) coincided exactly with wave V in the contralateral BAEP (maximal field amplitude of the order of 2-3 fT). In one subject, also a wave preceding wave V was observed, which was associated with basically the same spatial pattern as wave V itself, but had an opposite polarity. The study suggests that the supplementary information provided by BAEF measurements could be decisive for a better understanding of human auditory evoked brain stem activity. PMID- 10790853 TI - PLC beta 4-independent Ca2+ rise via muscarinic receptors in the mouse suprachiasmatic nucleus. AB - Cholinergic regulation of the suprachiasmatic nucleus (SCN) has been extensively studied although the intracellular signaling mechanisms are not well understood. We examined immunostaining for phospholipase C-beta (PLC-beta) families that couple to muscarinic acetylcholine receptors (mAChR) and demonstrated the expression of PLC-beta 1 and beta 4 in the mouse SCN. Ca2+ imaging analysis indicated that the MI-mAChR antagonist, pirenzepine blocked carbachol-induced Ca2+ elevation in the SCN and the response was equivalent between the wild type and the PLC-beta 4-knockout mice. In addition, the knockout mice displayed locomotor and temperature rhythms coupling to 24 h light/dark cycles. Therefore, it was proposed that PLC-beta 1 but not PLC-beta 4 was involved in the mAChR mediated Ca2+ signaling in the SCN. PMID- 10790855 TI - Frontal processing and auditory perception. AB - Disordered processing of the pattern in sound over time has been observed in a number of clinical disorders, including developmental dyslexia. This study addresses the brain mechanisms required for the perception of such a pattern. We report the systematic evaluation of temporal perception in a patient with a single intact right auditory cortex and a large right frontal lobe lesion. A striking dissociated deficit was demonstrated in the perception of temporal pattern at the level of tens or hundreds of milliseconds. This proves that, contrary to common belief, mechanisms in the pathway up to and including the primary auditory cortex are not sufficient for the normal perception of temporal pattern. This work suggests a need for frontal processing for the normal perception of auditory pattern. PMID- 10790856 TI - Treatment with delta opioid peptide enhances in vitro and in vivo survival of rat dopaminergic neurons. AB - A major problem in neural transplantation therapy is poor survival of grafted cells, which may be due to low cell viability prior to transplantation or scarce trophic factors available to the cells following transplantation. Recently, the delta enkephalin analogue [D-Ala(2),D-Leu(5)]-enkephalin (DADLE) has been demonstrated to protect against, as well as to reverse methamphetamine-induced loss of dopamine transporters. Here, we show that pretreatment with DADLE (0.0025, 0.005, 0.01 g/ml) dose-dependently enhanced cell viability of cultured primary rat fetal mesencephalic cells. In addition, DADLE administration in adult rats (4 mg/kg every 2 h, 4 injections, i.p.) prior to 6-hydroxydopamine lesions of the medial forebrain bundle, significantly reduced the severity of loss of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra 1 month post lesion. This is the first report suggesting that DADLE can be used as a supplement factor for improving the cell viability of fetal mesencephalic cells and as a protective agent against neurotoxicity in a Parkinson's disease model. PMID- 10790857 TI - Axonal transport of TREK and TRAAK potassium channels in rat sciatic nerves. AB - The recent cloning, functional expression and brain localization of two new potassium channels, TREK and TRAAK, led us to examine whether both channels are present in peripheral nerves and can move along axons by means of axonal transport mechanisms. Using specific antibodies directed against TREK and TRAAK peptides, we found that immunoreactivity for both potassium channels accumulates above and below a ligature in rat sciatic nerves. The process was rapid and bidirectional suggesting that the channels are associated with vesicles. This represents the first report on the axonal transport of potassium channels. PMID- 10790858 TI - Differential vulnerability of primary cultured cholinergic neurons to nitric oxide excess. AB - Many neuronal nitric oxide synthase (nNOS)-expressing brain neurons, including some cholinergic populations, are resistant to disease or to certain forms of excitotoxicity. Vulnerability to NO excess of forebrain (medial septal/diagonal band; MS-ACh) and brainstem (pedunculopontine/laterodorsal tegmental nuclei; BS ACh) cholinergic neurons was compared in E16-E18 primary rat brain cultures. MS ACh cells were approximately 300-fold more sensitive to the NO donor S-nitro-N acetyl-D,L-penicillamine (SNAP) than were BS-ACh cells. Most (69%) MS-ACh cells contained nuclear DNA fragments by 2 h after addition of SNAP, while only 21% BS ACh cells were TUNEL-positive after NO excess. Depletion of glutathione content did not potentiate the effect of SNAP on MS-ACh cells, but sensitized BS-ACh cells to the NO donor. Caffeic acid, a putative NF-kappa B inhibitor, enhanced the toxicity of SNAP to cholinergic neurons in both preparations. Our experiments show that cholinergic neurons in mixed primary cultures from different brain regions possess biochemical differences with respect to their vulnerability to NO excess. PMID- 10790859 TI - Amyloid beta-protein length and cerebral amyloid angiopathy-related haemorrhage. AB - The relationship between amyloid beta-protein (A beta) length and the apolipoprotein E (APOE) epsilon 2 allele, which is over-represented in cerebral amyloid angiopathy-related haemorrhage (CAAH), has not previously been examined. Of 57 CAA patients studied, 37 had CAAH. All patients, particularly those with CAAH had more blood vessels immunoreactive for A beta 40 than A beta 42 in both the leptomeninges and cerebral cortex. CAAH patients had more A beta 40 immunoreactive blood vessels in the leptomeninges (p < 0.001) and cortex (p = 0.027) than had non-haemorrhage patients. Cortical blood vessels, the usual source of haemorrhage in CAAH, were more frequently A beta 42 immunoreactive in APOE epsilon 2 carriers than in non-epsilon 2 carriers (p = 0.022). The APOE epsilon 2 allele may predispose to CAAH by increasing the seeding of cortical blood vessels by A beta 42. PMID- 10790860 TI - GABAB-RI receptors in serotonergic neurons: effects of baclofen on 5-HT output in rat brain. AB - The activation of GABAB receptors hyperpolarizes 5-HT neurons and reduces cell firing. In situ hybridization showed the presence of the GABAB-RI receptor transcript in virtually all 5-HT neurons of the dorsal and median raphe nuclei (DR and MnR, respectively) whereas the GAD transcript was present mainly outside these nuclei. The systemic administration of baclofen increased the in vivo 5-HT release in the DR, MnR and several projection areas. As shown previously in the DR, the application of baclofen in the MnR increased the local 5-HT output. Thus, although 5-HT neurons contain inhibitory GABAB-RI receptors, baclofen increased 5 HT release in some brain areas, likely by a preferential action on terminal GABAB autoreceptors in inhibitory inputs to 5-HT neurons. The scarcity of GAD expressing cells in the DR and MnR suggests that these inputs originate mainly outside these nuclei. PMID- 10790861 TI - 7-Nitroindazole blocks nicotine-induced conditioned place preference but not LiCl induced conditioned place aversion. AB - We have shown previously that the neuronal nitric oxide synthase (nNOS) is involved in the rewarding effect of cocaine as determined by the conditioned place preference (CPP) paradigm. In the present study we investigated the effect of the nNOS inhibitor 7-nitroindazole (7-NI) on nicotine-induced CPP and LiCl induced conditioned place aversion (CPA) in Swiss Webster mice. Mice treated with nicotine (0.5 mg/kg) or saline every other day for 8 days (four drug and four saline sessions) developed CPP that was completely blocked by pretreatment with 7 NI (25 mg/kg). Mice treated with LiCl (150 mg/kg) developed marked aversion to the LiCl-paired compartment. LiCl-induced CPA was not affected by the pretreatment with 7-NI. These findings suggest that nitric oxide (NO) plays a role in the acquisition of reward but not of aversion and that the blockade of nicotine-induced CPP is probably not due to impairment of learning and memory. PMID- 10790862 TI - Phosphorylation and destabilization of human period I clock protein by human casein kinase I epsilon. AB - Period (PER), a central component of the circadian clock in Drosophila, undergoes daily oscillation in abundance and phosphorylation state. Here we report that human casein kinase I epsilon (hCKI epsilon) can phosphorylate human PER I (hPER I). Purified recombinant hCKI epsilon (but not a kinase negative mutant of hCKI epsilon, hCKI epsilon-K38R) phosphorylated hPER I in vitro. When co-transfected with wild-type hCKI epsilon, in 293T cells, hPER I showed a significant increase in phosphorylation as evidenced by a shift in molecular mass. Furthermore, phosphorylation of hPER I by hCKI epsilon caused a decrease in protein stability in hPER I. Whereas phosphorylated hPER I had a half-life of approximately 12 h, unphosphorylated hPER I remained stable in the cell for > 24 h. hPER I protein could also be co-immunoprecipitated with transfected hCKI epsilon as well as endogenous hCKI epsilon, indicating physical association between hPER I and hCKI epsilon proteins in vivo. PMID- 10790863 TI - Activation of metabotropic GABAB receptors inhibited glutamate responses in spiral ganglion neurons of mice. AB - The lateral olivocochlear efferent (LOC) fibers form axodendritic synapses with auditory nerve dendrites synapsing on inner hair cells to affect postsynaptic responses GABA is one of the primary transmitters used by the LOC efferent. However, the molecular nature and developmental expression of the GABA receptors in spiral ganglion (SG) neurons are poorly understood. We have identified the GABAB receptors in SG neurons of mice by pharmacological criteria and polymerase chain reaction (PCR) detection. Applications of GABA or baclofen increased intracellular Ca2+ concentrations and inhibited glutamate responses. The responses were blocked by saclofen, but not by bicuculline. Relative-quantitative PCR showed that GABAB receptors were expressed prenatally, and their levels did not change significantly during the maturation of SG neurons. PMID- 10790864 TI - Expression of IL-6 in the ischemic penumbra. AB - We examined the expression of IL-6 within the ischemic penumbra at various time points after transient (3 h) middle cerebral occlusion (MCA-O) in rats. The animals were killed at 1, 3, 7 or 14 days following operation. Coronal brain sections were processed for immunohistochemistry with antibodies against GFAP, OX 42, IL-6 and Nissl-staining. Glial activation within the penumbra started on day one after ischemia and persisted up to day 14. Expression of IL-6 was not present in sham-operated controls. One day after MCA-O there were several IL-6-positive cells in the penumbra. This expression of IL-6 increased on day 3 and remained elevated up to day 14. According to the shape of the IL-6-positive cells they seem to be microglia and neurons. The present results demonstrate a longlasting expression of IL-6 in the ischemic penumbra. PMID- 10790865 TI - Electrophysiological studies in spinocerebellar ataxia type 6: a statistical approach. AB - In spinocerebellar ataxia type 6 (SCA6), the cerebellum is predominantly affected, but several electrophysiological studies have revealed subclinical disorders other than cerebellar lesions. We conducted statistical analyses by comparing SCA6 patients and age-matched normal controls to asses whether electrophysiological abnormalities are directly associated with SCA6 because late onset of SCA6 may involve senile changes. We performed brain stem auditory evoked potentials (BAEP), visual evoked potentials, somatosensory evoked potentials and nerve conduction studies in 10 SCA6 patients. The BAEP latencies of wave I was prolonged and compound muscle action potentials of peroneal nerve and sensory nerve action potentials of sural nerve reduced in SCA6 patients. Our results suggest an existence of peripheral impairment in the auditory pathway and axonal neuropathy in SCA6. PMID- 10790866 TI - Interactions of inflammatory mediators and low pH not influenced by capsazepine in rat cutaneous nociceptors. AB - The rat skin-saphenous nerve preparation was used to record from mechano-heat sensitive C-fibers whose receptive fields were superfused with various solutions of low pH and of bradykinin, serotonin and prostaglandin E2. Only synchronous application of protons and mediators resulted in a significant nearly three-fold augmentation of the nociceptive pH response, and capsazepine (10(-5) M) did not block this short-lived enhancement. Thus, it does not seem to involve the capsaicin receptor (VRI) which is in contrast to a previous finding from cultured sensory neurons. PMID- 10790867 TI - ERPs elicited by a cognitive incongruity paradigm: a semantic memory study. AB - The cognitive incongruity paradigm consists in presenting congruous or incongruous factual information to subjects, who are instructed to judge the truth of this information by comparing it with the factual knowledge stored in their semantic long-term memory. The factual information tested was whether a city presented after the name of a country, belongs (congruity) or not (incongruity) to that country. Results revealed that the recognition of incongruous factual information is characterized by a negative wave (N400), while the processing of congruous factual knowledge is characterized by a positive wave (P300) about 600 ms post-stimulus. In addition, these two components were sensitive to subject performance and subject confidence levels. The neurophysiological brain pattern observed during the cognitive incongruity paradigm reveals that the N400 and P300 are jointly affected by the task and reflect the recognition processes of factual knowledge. PMID- 10790868 TI - Increased levels of expression of an NMDARI splice variant in the superior temporal gyrus in schizophrenia. AB - Expression patterns of mRNAs for the NMDARI subunit (NRI) carboxy-terminus isoforms were investigated in postmortem brain tissue using isotopic in situ hybridization. Three brain regions (superior temporal, middle frontal and visual cortices) were examined in patients with schizophrenia (n = 6) and control subjects (n = 6). A 22% higher level of expression of the NRI isoform that contains neither spliced exon was observed in the superior temporal gyrus of patients with schizophrenia compared with controls (p = 0.01). No differences were observed in the expression of the other isoforms in the three regions studied. These data suggest that NRI alternative splicing might be abnormal in schizophrenia and reinforce previous findings implicating the superior temporal gyrus as a site of neural dysfunction in schizophrenia. PMID- 10790870 TI - P2X3 receptor in injured human sensory neurons. AB - The ATP-gated cation channel P2X3 is expressed selectively by rat sensory neurones, and may play a role in nociception by binding ATP released from damaged or inflamed tissues. However, the distribution of this channel in human sensory neurons is not known. Using a specific antibody, we have demonstrated intense P2X3 immunoreactivity within a subset (60%) of small/medium diameter sensory neurones and fine nerve fibres in intact post-mortem human dorsal root ganglia (DRG). Co-localization studies showed < 15% overlap with the trkA immunostaining in DRG, indicating that P2X3 was expressed predominantly in sensory neurons that are also isolectin B4 positive. There was a significant decrease in numbers of P2X3-like immunoreactive neurons in human DRG after central axotomy (to 36%), similar to the decrease in rat DRG after peripheral axotomy. However, Western blotting demonstrated a specific 66 kDa band in human DRG and peripheral organs, including intestine, where histochemistry showed P2X3 immunoreactivity in myenteric plexus neurons. Thus P2X3 antagonists may be analgesic, but are unlikely to have a selective effect on pain in humans. PMID- 10790869 TI - Tracking Alzheimer's disease in transgenic mice using fluorodeoxyglucose autoradiography. AB - While transgenic mice have great promise in the study of Alzheimer's disease (AD), uncertainties remain about the extent to which they provide a model of the disorder or the best way to characterize disease progression. Using fluorodeoxyglucose (FDG) autoradiography, we found that transgenic mice over expressing a mutant form of the human amyloid precursor protein have preferentially and progressively reduced activity in the posterior cingulate cortex and relatively spared activity in visual cortex, sensorimotor cortex, cerebellum and brain stem, a pattern previously demonstrated in FDG PET studies of persons with Alzheimer's disease, Brain imaging of posterior cingulate activity could provide an indicator of AD in suitable animals, helping to clarify disease mechanisms and screen candidate treatments. PMID- 10790871 TI - High-frequency synaptic stimulation induces association of fyn and c-src to distinct phosphorylated components. AB - Signaling via tyrosine kinases appears necessary for regulation of synaptic efficacy. Interactions of the src-family kinases with phosphorylated proteins were studied in area CAI of rat hippocampal slices 10 min after induction of long term potentiation (LTP) by 100 Hz/l s stimulation (HFS). HFS enhanced association of the src-family kinases fyn and c-src with an approximately 120 kDa tyrosine phosphorylated component containing the focal adhesion kinase (FAK) and its homologue PYK2. Association of fyn with FAK and of c-src with PYK2 was increased following the HFS. Further, increase in tyrosine phosphorylation of PYK2 was detected following the HFS. These results suggest that fyn and c-src are involved in distinct signaling pathways and provide evidence for activation of FAK and PYK2 following synaptic stimulation inducing LTP in vitro. PMID- 10790872 TI - Transient over-expression of NGFI-A gene suppresses NGF-induced neurite outgrowth in PCI2 cells. AB - Using an inducible gene expression system (Tet-ON system), the role of NGFI-A gene during the neuronal differentiation of PCI2 cells was examined. When NGFI-A was transiently over-expressed, no obvious effects on cell proliferation or neurite outgrowth were observed. Interestingly, however, NGFI-A over-expression resulted in significant retardation in NGF-induced neurite outgrowth. Similar suppressive effects were observed also on the v-K-ras-induced neurite outgrowth. These results raise the possibility that NGFI-A protein may play some negative role in NGF signaling. PMID- 10790873 TI - Changes in cortical activity during suppression in stereoblindness. AB - Patients with strabismus or anisometropic amblyopia fixate and attend with one eye and suppress the image from the other eye. Here we use a visual evoked potential technique to show that patients who lack normal stereopsis retain suppressive binocular interactions but lack a characteristic form of non-linear binocular interaction that is present in normal observers. Oscillating grating targets presented at different temporal frequencies in the two eyes evoke a strong response in normal observers at a frequency equal to the sum of the two input frequencies for fusable targets but not for rivalrous ones. However increasing contrast in one eye reduces the response amplitude from the other eye under either fusable (dichoptic masking) or rivalrous conditions. Stereo deficient observers lack the sum-frequency response, but retain dichoptic masking interactions. Dichoptic masking is stronger when the masker is presented to the patients' dominant rather than non-dominant eyes, suggesting that a subset of preserved binocular inhibitory interactions form the basis of clinical suppression. PMID- 10790874 TI - Redistribution of bisbenzimide Hoechst 33342 from transplanted cells to host cells. AB - Identification of transplanted cells within host tissue is an important component of many transplantation experiments. In this study, Schwann cells labelled with the fluorochrome bisbenzimide (Hoechst 33342, H33342) and transduced with the lac Z gene were introduced into normal white matter and their distribution was examined 2 h, 24 h and 4 weeks after transplantation. At 2 and 24 h following transplantation, H33342-labelled cells were more widely distributed than lac-Z labelled cells in both longitudinal and transverse directions. By 4 weeks following transplantation, no lac-Z-labelled cells could be found. However, H33342-labelled cells were observed in and around the glial scar. Therefore, labelling of host cells by transfer of H33342 dye from transplanted cells has to be considered whenever this dye is used as a transplant marker. PMID- 10790875 TI - Peripheral electric stimulation inhibits morphine-induced place preference in rats. AB - Conditioned place preference (CPP) is a commonly used model to detect rewarding effect of drugs. To observe the effect of peripheral electric stimulation (PES) on morphine-induced CPP, we trained the rats with morphine in a CPP paradigm. Twelve hours before the testing phase, rats were given PES via stainless-steel needles with frequencies of 2, 100, or 2/100 Hz, respectively. PES of 2 and 2/100 Hz significantly decreased CPP in morphine-trained animals in a naloxone reversible manner, while PES of 100 Hz, foot shock, needle insertion, or plain restraining, showed no effect. Thus, PES with a low-frequency component (2 Hz) could specifically inhibit the expression of morphine-induced CPP, presumably via activation of opioid receptors. PMID- 10790876 TI - Serotonin depletion decreases light induced c-fos in the rat suprachiasmatic nucleus. AB - The suprachiasmatic nucleus (SCN) is the locus of the biological clock in mammals. Daily light cycles entrain the endogenous circadian rhythms in mammals through direct and indirect neural pathways from the retinae to the suprachiasmatic nucleus. We have studied the effect of serotonin depletion on the photic induction of the early response gene c-fas in the SCN of rats. Serotonin depletion, verified by immunohistochemistry, produced a significant decrease (42%) in the number of c-FOS positive cells in the ventrolateral portion of the SCN. These results support the involvement of serotonin as a mediator of photic information to the SCN through the retinal projection to the dorsal raphe nucleus. PMID- 10790877 TI - Are A3 adenosine receptors expressed in the brain? AB - An increasing number of reports suggest a role for A3 adenosine receptors (A3ARs) in mediating adenosine action in the central nervous system. However, studies of A3AR localization in the brain have yet to be performed. To provide insights into the central sites of A3AR action, we compared patterns of A1 and A3AR mRNA and binding site expression in the brains of rats, mice and humans. We also assessed whether A3 agonists are selective for A3ARs. Whereas it was possible to detect high-level A1AR expression in many brain regions, it was not possible to detect either A3AR gene or binding site expression in the central nervous system. When we examined the affinities of the A3AR agonists CI-IAB-MECA and IAB-MECA for A1ARs, we found that these compounds bound to A1ARs with high affinity. These observations suggest that studies using A3-agonists need to consider potential effects of A1ARs activation, as A1ARs are abundantly expressed in the nervous system whereas A3AR expression in the brain cannot be directly demonstrated. PMID- 10790878 TI - Quantitative aspects of the state-dependent co-variation of cat lateral geniculate and perigeniculate visual activity. AB - The quantitative relationship between EEG-related changes in the visual activity of perigeniculate (PGN) and lateral geniculate (LGN) neurons with overlapping receptive fields was analyzed in the anesthetized and paralyzed cat. While transient response peaks were independent of the EEG state, we found opposite changes in spontaneous activity and tonic visual responses, with PGN cells increasing and LGN cells decreasing their spontaneous/tonic activity with increasing EEG delta activity in most cases. The tonic firing rates of PGN and LGN cell pairs were clearly correlated with a slope of about -0.5. Thus, LGN firing was low when PGN activity was high and vice versa. With a change from low to high EEG delta activity the difference between the tonic responses of PGN and LGN cells increased on average by 50 spikes/s, both for the whole population and at the single cell level, indicating that state-dependent changes in retino geniculate transmission are regulated by a distinct ratio of PGN-LGN activity. PMID- 10790879 TI - A lesion of cortical area V2 selectively impairs the perception of the direction of first-order visual motion. AB - Lesions of area MT/V5 in monkeys and its presumed homologue, the motion area, in humans impair motion perception, including the discrimination of the direction of global motion in random dot kinematograms. Here we report the results of similar tests on patient TF, who has a discrete and very small, unilateral infarct in the medial superior part of the right occipital cortex. Structural MRI, co-registered in software with a standardized human brain atlas, reveals that the lesion involves area V2. The patient was impaired in his retinotopically corresponding left lower quadrant on several motion tasks including discrimination in random dot kinematograms of direction, speed and motion-defined discontinuity. He was also impaired on tasks selectively involving first-order motion based on luminance contrast but not on second-order motion based on texture contrast. The results show that even though area MT/V5 is intact, motion perception is abnormal and, in particular, his perception of first-order motion is impaired. PMID- 10790880 TI - Population coding of movement dynamics by cerebellar Purkinje cells. AB - In order to accomplish desired movements, the nervous system must specify the movement dynamics: it must provide a signal that compensates for the mechanical constraints encountered during movement. Here we tested whether the population activity of Purkinje cells in the cerebellum specifies the dynamics for pursuit eye movements. We first estimated the population activity by computing weighted averages of Purkinje cell firing on a millisecond time scale. We then generated predicted eye movements by transforming this pooled neural activity with a description of eye mechanics. We found that the equally weighted average of Purkinje cell outputs produced a close match between the predicted and actual eye movements. These findings demonstrate that neural circuits through the cerebellum are capable of providing the dynamic compensation necessary to achieve desired movements. PMID- 10790881 TI - Olfactory based spatial learning in neonatal mice and its dependence on CaMKII. AB - Spatial learning and memory involves the ability to encode geometric relationships between perceived cues and depends critically on the hippocampus. Visually guided spatial learning has been demonstrated in adult animals. As infant animals rely heavily on olfaction, olfactory based spatial learning was assessed in infant mice. When 12-day-old pups were displaced from their nest, they learned within a few training trials to use the spatial pattern of odor cues to move back to the nest. However, mouse pups that over-expressed Ca2+/calmodulin dependent protein kinase (CaMKII) in hippocampal neurons were impaired in olfactory based spatial learning. PMID- 10790882 TI - Methamphetamine induces cytosolic Ca2+ oscillations in the VTA dopamine neurons. AB - Methamphetamine (METH) induces a schizophrenia-like psychosis. The dopamine neurons in the ventral tegmental area (VTA) have been implicated in schizophrenia and drug abuse. The present study investigated direct effects of METH on VTA dopamine neurons. We treated adult SD rats with METH (5 mg/kg/day) or saline for 7 days, isolated single VTA neurons, and monitored neuronal activities by measuring cytosolic Ca2+ concentration ([Ca2+]i) in immunocytochemically identified dopamine neurons. Acutely administered METH increased [Ca2+]i in dopamine neurons from METH- and saline-treated rats and induced oscillations of [Ca2+]i in dopamine neurons only from METH-treated rats. The METH-induced [Ca2+]i oscillations were inhibited by Ca(2+)-free conditions and Ca2+ channel blockers. The results indicate that acute METH increases [Ca2+]i in VTA dopamine neurons and that subchronic METH treatment sensitizes them to this drug, resulting in induction of [Ca2+]i oscillations. The activation of VTA dopamine neurons may be related to psycho-stimulant effects of METH. PMID- 10790883 TI - Chondroitinase ABC promotes axonal regeneration of Clarke's neurons after spinal cord injury. AB - We examined whether enzymatic digestion of chondroitin sulfate (CS) promoted the axonal regeneration of neurons in Clarke's nucleus (CN) into a peripheral nerve (PN) graft following injury of the spinal cord. After hemisection at T11, a segment of PN graft was implanted at the lesion site. Either vehicle, brain derived neurotrophic factor (BDNF) or chondroitinase ABC was applied at the implantation site. The postoperative survival period was 4 weeks. Treatment with vehicle or BDNF did not promote the axonal regeneration of CN neurons into the PN graft. Application of 2.5 unit/ml chondroitinase ABC resulted in a significant increase (12.8%) in the number of regenerated CN neurons. Double labeling with Fluoro-Gold and NADPH-diaphorase histochemistry showed that the regenerated CN neurons did not express nitric oxide synthase (NOS). Our results suggest that CS is inhibitory to the regeneration of CN neurons following injury of the spinal cord. PMID- 10790884 TI - Cortical development and saccade planning: the ontogeny of the spike potential. AB - The spike potential is a sharply timed positivity which precedes eye movements in adults, and is thought to indicate cortical planning of saccades. While the spike potential is observed under most conditions in adults, it has not been reported in young infants. In the present study we shed light on the ontogeny of the spike potential by demonstrating for the first time its existence in a group of older infants (12 months). This result is consistent with a relatively delayed onset of cortical control over saccades during development. PMID- 10790885 TI - Novel expression of hypothalamic neuropeptide Y during postnatal development in the rat. AB - Neuropeptide Y (NPY) is a potent orexigenic peptide. In the normal adult rat, hypothalamic NPY mRNA expression is limited to the arcuate nucleus (ARH). The purpose of this study was to characterize the developmental expression of NPY mRNA in the hypothalamus of the rat. In contrast to the normal adult rat, NPY mRNA was observed in the ARH, the dorsomedial hypothalamic nucleus (DMH) and the perifornical region (PFR) during development. NPY mRNA expression in all three regions increased progressively from postnatal days 0-4 (P0-4) to reach maximum levels at P16 and subsequently decreased to near adult levels by P30. The unique expression of NPY mRNA in the PFR and DMH may be important in establishing the proper management of energy homeostasis and body weight in the adult animal. PMID- 10790886 TI - Neurons and ECM regulate occludin localization in brain endothelial cells. AB - We report that extracellular matrix and neurons modulate the expression of occludin, one of the main components of tight junctions, by rat brain endothelial cells (RBE4.B). Of the three extracellular matrix proteins which we tested (collagen I, collagen IV, and laminin), collagen IV stimulated at the best the expression of occludin mRNA. The corresponding protein, however, was not synthesized. Significant amounts of occludin accumulated only when RBE4.B cells were cultured on collagen IV-coated inserts, in the presence of cortical neurons, plated on laminin-coated companion wells. Finally, occludin segregated at the cell periphery, only when endothelial cells were co-cultured with neurons for > or = 1 week. PMID- 10790887 TI - Heroin-induced neuronal activation in rat brain assessed by functional MRI. AB - The present study demonstrates the application of fMRI technology to neuropharmacology and the interaction of drug/receptor in the rat brain. Specifically, we have observed two different types of fMRI signal changes induced by acute i.v. heroin administration in rat brains under conditions of spontaneous and artificial respiration. Under spontaneous respiration, a global decrease in fMRI signal was observed; under artificial respiration, a region-specific increase in fMRI signal was identified and the activation sites are consistent with the distribution of opiate mu-receptors in rat brain as previously reported by autoradiography. Both heroin-induced fMRI signal changes were suppressed by pretreatment of naloxone, an opiate mu-receptor antagonist, and reversed by injection of naloxone following heroin infusion. These results suggest that fMRI has specific advantages in spatial and temporal resolution for studies of neuropharmacology and drugs of abuse. PMID- 10790888 TI - Evidence for resistance to MPTP in C57BL/6 x BALB/c F1 hybrids as compared with their progenitor strains. AB - The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is capable of producing a syndrome in mice which shares major characteristics with human Parkinson's disease. There is evidence for a genetic influence on the degree of damage exerted by MPTP, since different strains of mice can dramatically differ in their response to MPTP. We produced reciprocal F1 hybrids by crossbreeding the MPTP-susceptible C57BL/6 strain with resistant BALB/c. These hybrids were compared to the parental strains using neural and behavioral measures in order to characterize the genetic transmission of MPTP-susceptibility. The F1 generation as a whole had a lower depletion of neostriatal dopamine levels than even found in BALB/c. Furthermore, there was no significant loss of tyrosine hydroxylase positive cells in the substantia nigra and quick recovery from deficits in motor behavior in F1, herein resembling BALB/c. We suggest that several loci are involved in susceptibility to MPTP, and that the trait is under control of recessive susceptibility and/or dominant resistance alleles, which interact in F1, leading to extremely low susceptibility. PMID- 10790889 TI - Multiple time-frequency components account for the complex functional reactivity of P300. AB - Consecutive and overlapping time-frequency (TF) components of auditory event related brain potentials (ERPs) were analyzed to examine whether multiple co existing components may account for the complex functional reactivity of P300. Auditory ERPs of 14 adult subjects were decomposed by means of the wavelet transform (WT), and TF components within P300 were tested in a systematic manner for the effects of major P300 determinants: stimulus probability, active discrimination, and mental count task. The results demonstrated that several partly or fully simultaneous delta, theta, and alpha TF components significantly depend on the factors eliciting P300, and also manifest distinct patterns of task reactivity and scalp distribution. Thus, specific functional processes that underlie the P300 ERP can be distinguished that help to account for its responsiveness to task variables. PMID- 10790890 TI - Glutamate release and neuronal injury after intrathecal injection of local anesthetics. AB - High concentrations of local anesthetics are neurotoxic, but the mechanism for this neurotoxicity is obscure. Here, we report increased concentrations of glutamate in the cerebrospinal fluid after intrathecal injections of high concentrations of tetracaine (a local anesthetic). The peak concentrations of glutamate after administration of 1%, 2%, and 4% tetracaine were 4-fold, 6-fold, and 10-fold higher than baseline values, respectively. Animals in the 1% group were all neurologically normal one week after tetracaine injection. In the group receiving 4%, no animal was able to hop and vacuolation of the white matter and/or central chromatolysis of the motor neurons were observed. Because high concentrations of glutamate are known to be neurotoxic, our results may provide some insight into the mechanisms for neurotoxicity of intrathecal local anesthetics. PMID- 10790891 TI - Expression of nerve growth factor-induced type 1 plasminogen activator inhibitor (PAI-1) mRNA is inhibited by genistein and wortmannin. AB - Nerve growth factor (NGF), which acts as a neurotrophic factor in a rat pheochromocytoma cell line (PC12), stimulated type 1 plasminogen activator inhibitor (PAI-1) mRNA expression from 1 to 5 h, after addition at 5 ng/ml. PAI-1 antigen in culture medium, which was measured using an enzyme linked immunosorbent assay (ELISA), was also increased dose dependently by the addition of NGF. Neither epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), phorbol myristate acetate (PMA) nor forskolin increased PAI-1 mRNA expression in PC12 cells. Genistein, an inhibitor of tyrosine protein kinase, completely inhibited NGF induced PAI-1 mRNA in the presence of 100 microM. Wortmannin, a potent and specific inhibitor of phosphatidylinositol 3-kinase (PI 3 kinase), decreased induction of PAI-1 mRNA level at doses of > or = 10(-7) M. PMID- 10790892 TI - Selective vulnerability of spinal cord motor neurons to non-NMDA toxicity. AB - We previously reported that alpha-motor neurons in organotypic cultures of rat spinal cord (OTC-SC) are resistant to excitotoxicity induced through NMDA receptors. Here we describe the effects of non-NMDA glutamate receptor agonists kainic acid (KA) and quisqualic acid (QUIS) on motor neurons in OTC-SC. Large ventral horn acetylcholinesterase-positive neurons (VHANs), most of which are motor neurons, were quite sensitive to QUIS and KA toxicity and displayed losses of 95% and 94%, respectively. Small VHANs were reduced by 41% and 61% only. Identical results were obtained in cultures stained for non-phosphorylated neurofilaments. These observations demonstrate that alpha-motor neurons are considerably more sensitive to KA and QUIS than to NMDA toxicity. The proposed excitotoxic mechanism of ALS, therefore, is most likely mediated through non-NMDA glutamate receptors. PMID- 10790894 TI - Dissociable ERP profiles for processing rules vs instances in a cognitive sequencing task. AB - A hotly debated question in cognitive neuroscience is whether individual instances of perceptual sequences, and the rules that describe them, are processed by the same brain mechanisms. We tested the hypothesis that such rules and instances are processed by dissociable brain mechanisms. We analyzed event related brain potentials (ERPs) evoked during cognitive sequencing tasks that assessed surface (instance) vs abstract (rule) structure learning. Sequence instances ABCBAC and DEFEDF have different serial order or surface structure, but share the same rule or abstract structure, 123213. Nine healthy subjects were first trained to learn a set of surface and abstract structures in sequences of visually presented stimuli. During the subsequent ERP recording, for surface and abstract structures, they then discriminated between acceptable and unacceptable sequences, based on the pre-learned regularities. Abstract structure processing evoked a late positivity around 500 ms, which was not seen in the surface structure processing, supporting our hypothesis of dissociable processes. We discuss implications for the rule vs instance debate, and similarities between this late positivity and the P600 observed in previous studies of syntactic processing. PMID- 10790893 TI - In vitro differentiation of multipotent human neural progenitors in serum-free medium. AB - Stem cells are exciting candidates for therapeutic strategies in neurodegenerative diseases, due to their multipotency and migratory capabilities. We show that stem cell-like embryonic normal human neural progenitors (HNPs) are capable of proliferating in response to mitogenic growth factors and differentiate into diverse CNS cell types in vitro. We present evidence that HNPs differentiate to beta III-tubulin-, glial fibrillary acidic protein- and O4 immuno-positive cells, in both a fetal bovine serum-containing and a non supplemented, serum-free basal medium. From these findings, we propose that HNPs may differentiate not only in response to exogenous differentiation factor(s) contained in the serum, but also in response to some endogenous factor(s) released from the HNPs, which may regulate the differentiation pathway of these cells. PMID- 10790895 TI - Polymorphisms of genes controlling homocysteine/folate metabolism and cognitive function. AB - Elevated concentrations of the amino acid homocysteine and/or folate deficiency have been reported to affect neural development/function in both human patients and animal models. We have investigated the distribution of functional polymorphisms in genes involved in homocysteine/folate metabolism in children with high IQ and in children with average IQ. No differences in the frequencies of genetic variants in the methionine synthase or methylenetetrahydrofolate reductase genes were found. However, the cystathionine beta-synthase (CBS) 844ins68 allele was significantly underrepresented in children with high IQ. The mechanism by which a functional genetic variant in the CBS gene may influence cognitive function remains to be determined. PMID- 10790896 TI - GABA-A antagonist causes dramatic expansion of tuning in primary auditory cortex. AB - Responses from 80 neurons in primary auditory cortex of anesthetized chinchillas were investigated before and during iontophoresis of bicuculline (BIC), a GABA-A antagonist. BIC caused a significant increase in the spontaneous discharge rate (SR) and driven firing rate in most neurons. Threshold decreased in many neurons and the excitatory response area expanded significantly above and below the neuron's characteristic frequency (CF). These results indicate that many neurons in primary auditory cortex receive excitatory inputs from a much broader frequency range than previously believed. GABA-A mediated inhibition can significantly alter the excitability and tuning of auditory cortex and could contribute significantly to cortical plasticity due to experience and cochlear pathology. PMID- 10790897 TI - Strange bedfellows: a critical view of pathological gambling and addiction. PMID- 10790898 TI - Community effects of the opening of the Niagara casino. AB - AIMS: The impacts on the community of the opening of a casino in Niagara Falls are studied. DESIGN: The study uses a pre/post design for the community data, with pre/post data from Ontario as a whole as a comparison. SETTING: The study site is the city of Niagara Falls, Ontario, where a casino opened in early December, 1996. PARTICIPANTS: Using random-digit dialing, telephone interviews were conducted with adult residents of Niagara Falls in 1996 and 1997, and with adult residents of Ontario in 1995 and 1997. MEASUREMENTS: Aside from demographic variables, measures included general attitudes to gambling, expectations about (1996) and experiences with (1997) the casino's opening, extent of participation in 11 types of gambling and 18 items on problems with gambling: five key items from a standard gambling problems score (SOGS), five life-area problems items, and items on pressures from others concerning the respondent's gambling and on gambling problems among family and friends. FINDINGS: Attitudes to gambling remained stable in Niagara Falls, while there was some evidence of decline in approval in Ontario as a whole. While strong majorities of 1996 respondents had expected many positive and negative effects on The Community of the Casino's opening, Significantly fewer respondents in 1997 reported actually experiencing most of these effects. While a small increase in employment was found, it fell far below projections, a result probably reflecting displacement effects. The rate and level of casino gambling increased in Ontario, but increased even more in Niagara Falls, with little displacement of other gambling. Reported gambling problems increased significantly in Niagara Falls for two of 10 gambling problem items and for the short SOGS score, while rates were generally stable or declining in the province. Pressure from others about gambling rose significantly in Niagara Falls (in contrast to the province), and reported rates of family members or friends with gambling problems also rose substantially. There was an increasing trend in Niagara Falls for all 18 problem indicators. CONCLUSIONS: The casino's opening brought more gambling by local residents, and an increase in reported gambling problems; yet support for the casino, already strong, if anything grew. At least in the short term, problems from the increased availability of gambling manifested themselves not in the public arena but rather in the arena of private life. PMID- 10790899 TI - Alcohol consumption and alcohol-related problems: Chinese experience from six area samples, 1994. AB - AIMS: To understand the drinking frequency, mean yearly volume of consumption and drinking-related problems in individuals living in six areas in China. DESIGN AND MEASUREMENTS: By two-stage sampling, 23,513 community household residents (15-65 years old) were assessed with data collected by trained psychiatrists using structured questionnaires and DSM-III-R criteria. FINDINGS: The male, female and total drinking rates were 84.1%, 29.3% and 59.5%, respectively. More than half of male drinkers and 90.0% of female drinkers used alcohol once a week or less. More than 16.1% of male drinkers and 2.5% of female drinkers drank once a day or more. The males reported a greater amount of alcohol consumed per session. The mean yearly volume of consumption of pure alcohol per year for all sample was 3.60 l; males consumed 18.6 times more alcohol than females. The rate of alcohol-related problems was higher in males than females. The male, female and total alcohol dependence point prevalence rates were 6.632%, 0.104% and 3.428%, respectively. Six-month incidence rates of acute intoxication were 5.162% 0.017% and 2.637%, respectively. CONCLUSION: Drinking problems in China will continue to be a public health problem in the next century. PMID- 10790900 TI - Heavy alcohol consumption and marital status: disentangling the relationship in a national study of young adults. AB - AIMS: To investigate why alcohol consumption varies by marital status, assessing (i) differences in heavy consumption prior to changes in marital status (indicating selection) and increases or decreases in heavy consumption associated with changes in marital status (indicating causation), (ii) whether such increases or decreases are transient, and (iii) the possible mediating effect of parental status. DESIGN: Longitudinal cohort. SETTING: Great Britain. PARTICIPANTS: Data from the 23- and 33-year surveys of the 1958 British birth cohort (all born in England, Wales and Scotland, 3-9 March 1958). MEASUREMENTS: Heavy drinking, defined as more than 35 (men) and 20 (women) units/week; changes between ages 23 and 33 in consumption and marital status. FINDINGS: The divorced had the highest consumption levels at both ages, the married had the lowest. Selection effects were minimal in both sexes. Overall, heavy drinking declined between ages 23 and 33 (21.4-13.0% in men, 6.4-3.4% in women), but increased among individuals who divorced, compared to the continuously married (adjusted OR = 2.05, 95% CI = 1.49, 2.83 for men; OR = 2.61, 95% CI = 1.67, 4.09 for women), most strikingly for recent divorces (adjusted OR = 4.97, 95% CI = 2.86, 8.57 and OR = 5.25, 95% CI = 2.60, 10.65). High rates of heavy drinking persisted for never married men (19.1%) and women (5.2%). CONCLUSIONS: The heavy drinking level of divorced young adults was not due to selection. Marital separation was accompanied by increases in heavy drinking, with pronounced short-term effects. Adverse alcohol-related health consequences may occur in the immediate period around divorce. Individuals who never marry appear to have a chronic heavy consumption pattern that may contribute to their increased mortality. PMID- 10790901 TI - Psychiatric disorders in adult children of problem drinkers: prevalence, first onset and comparison with other risk factors. AB - AIMS: (1) To confirm the increased risk of psychiatric disorders in Adult Children of Alcoholics (ACOAs); (2) to test if the age of onset of the disorders differs for ACOAs versus non-ACOAs; (3) to estimate the weight of being an ACOA compared to other risk factors including childhood traumas, other parental problem behaviours and current risk factors. DESIGN, SETTING, AND PARTICIPANTS: A random sample of the Dutch population (N = 7147) was interviewed (response rate: 69.7%). MEASUREMENTS: Psychiatric disorders were assessed using the CIDI. Parental problem drinking, other parental problem behaviours and childhood traumas were assessed using self-report measures. FINDINGS: (1) ACOAs had a significantly higher life-time, 12-month and 1-month prevalence of mood, anxiety and abuse/dependence disorders. Sons of problem drinkers also had a higher prevalence of eating disorders and schizophrenia. The prevalence rates were particularly high for the children of fathers with drinking problems. (2) The first onset of the mood and anxiety disorders took place at a younger age in ACOAs than in non-ACOAs. (3) Relative to other parental problem behaviours and childhood traumas, parental problem drinking is a strong predictor of psychiatric disorders, in particular abuse/dependence disorders. CONCLUSIONS: Children of fathers with a drinking problem are a high-risk group for psychiatric disorder. From a public health perspective it is an important target to break through this continuing circle. The further development of prevention and early treatment interventions at schools, youth care and addiction treatment centres is an important issue. PMID- 10790902 TI - Brief intervention for alcohol use in pregnancy: a randomized trial. AB - AIMS: To assess the impact of a brief intervention on antepartum alcohol consumption. DESIGN: A randomized clinical trial. SETTING: The obstetrics practices of the Brigham and Women's Hospital in Boston, MA, USA. PARTICIPANTS: Two hundred and fifty eligible women initiating prenatal care. INTERVENTION: A comprehensive assessment of alcohol use (assessment only, AO) or the same comprehensive assessment with a brief intervention (BI). MEASUREMENT: Demographic background and obstetric history of subjects, current and lifetime use of alcohol and substances, composite Addiction Severity Index scores, and antepartum alcohol use. FINDINGS: Of the 250, 247 (99%) subjects provided information on their antepartum drinking. Both the AO and BI groups had reductions in antepartum alcohol consumption, but differences in reductions by group were not statistically significant (p > 0.05). Risk of antepartum drinking after either the AO or BI was increased nearly threefold if the subject had any prenatal alcohol consumption before assessment (p = 0.0001). For the 143 subjects who were abstinent pre-assessment, however, those who received the BI maintained higher rates of abstinence (86% versus 72%, p = 0.04). CONCLUSIONS: After a comprehensive assessment of alcohol use, subjects in both the AO and BI groups reduced their antepartum alcohol consumption. The importance of screening for prenatal alcohol use is underscored by the findings that any prenatal alcohol consumption increases the risk of continued antepartum drinking. PMID- 10790903 TI - Quantitative and qualitative evaluations of brief interventions to change excessive drinking, smoking and stress in the police force. AB - AIMS: To evaluate the effects of a brief intervention to reduce excessive drinking, smoking and stress among police. DESIGN: (1) Controlled intervention trial with pre and post-intervention assessment approximately 8 months apart; (2) focus group identification of relevant factors). SETTING, PARTICIPANTS: Assessment was carried out of 954 NSW (Australia) police at 19 stations within two matched districts in the Sydney metropolitan area. Five focus groups were carried out with 43 randomly selected police from the matched districts. MEASUREMENTS: Weekly alcohol consumption and binge drinking, smoking and symptoms of stress were measured by a self-administered Health and Fitness Questionnaire. Recorded responses to set questions provided qualitative data. RESULTS: Participation was high (89%) at both quantitative assessments. Alcohol consumptions, particularly among men, was high at both baseline and follow-up assessments, although comparisons between groups across occasions showed no significant intervention effects. Excessive drinkers and those reporting moderate to severe stress levels reported more sick leave days (p < 0.05, p < 0.05). A significant increase in awareness of alcohol policies in the work-place showed in both experimental and control groups over time (p < 0.01). The percentage of smokers declined significantly in both intervention and control groups. Overall, women had significantly more symptoms of stress than men. Only 20% of police thought they would seek advice from work-place staff about alcohol consumption, 14% for smoking and 61% for stress. In the qualitative study, employees generally distrusted their organization's involvement in health unless work performance was affected. Seeking professional assistance for life-style issues was viewed as a sign of weakness. Alcohol use was seen as a way of obtaining information or group membership, self-medication and socializing. CONCLUSIONS: The brief interventions did not produce significant improvements in three life-style factors beyond positive trends in alcohol consumption among women and general reductions in smoking among both study groups. Combining quantitative and qualitative approaches helped identify interactive individual and organizational factors which influence behavioural and cultural norms. PMID- 10790904 TI - The use of acute and preventative medical services by a general population: relationship to alcohol consumption. AB - AIMS: To investigate the hypothesis that increasing alcohol consumption is accompanied by increasing use of acute, but decreasing use of preventative, medical services among the general population. DESIGN AND PARTICIPANTS: Health and life-style survey of 41,000 randomly-sampled adults in SE England who self completed a validated questionnaire covering socio-demographics, alcohol and tobacco usage and use of acute (A&E department and general practitioner) and preventative (dental, optician, mammography and cervical cytology) services: the response rate was 60%. MEASUREMENTS: Comparative use of acute and preventative health care services by patients with varying consumption of alcoholic beverages. This was estimated by the odds ratio for service use, after correcting for the following confounding variables; age, social class, ethnic group, employment status, whether lives with children or with other adults, whether is a career, limiting long-term illness, depression status, smoking habit and use of private health insurance. FINDINGS: There was increased use of accident and emergency services by the harmful and intermediate drinking groups compared with the safe drinking group. Male abstainers attended their A&E departments more frequently than 'safe limit' drinkers. With respect to preventative services, both male and female abstainers and harmful drinkers used dental services less than safe limit drinkers. For females, mammography and cervical cytology services were less frequently used by abstainers and by harmful drinkers. CONCLUSIONS: This study supports the generally held view that heavy alcohol consumers are disproportionate users of acute medical services but they are relative under users of preventative medical care services. Alcohol abstainers are also over users of acute services, but under-users of preventative services. These latter observations are relevant to the claims that moderate alcohol consumers have lower apparent morbidity and mortality rates compared to abstainers. PMID- 10790905 TI - Correlates of benzodiazepine abuse in methadone maintenance treatment. A 1 year prospective study in an Israeli clinic. AB - AIMS: This study addressed the following questions for patients after 1 year of methadone maintenance treatment (MMT); (1) What are the demographic features and past history of drug use of benzodiazepine (BZD) abusers? (2) Do BZD abusers abuse more heroin, cocaine and/or cannabis and do they receive a higher methadone dosage level? (3) Do BZD abusers suffer more from hepatitis C (HCV) and do they have more HIV/HCV risk-taking behaviors than non-abusers? (4) Do BZD abusers have more psychopathology and more emotional distress than non-abusers? DESIGN: All 148 patients who completed 1 year of MMT underwent random and twice-weekly observed urine analysis for various drugs of abuse, responded to self-report questionnaires (SCL-90-R; POMS; HIV/HCV risk-taking behaviors), interviews (ASI) and underwent testing for hepatitis C. Abuse in this study is defined as any use during the 12th month of treatment. FINDINGS: After 1 year of MMT, more BZD abusers (n = 63) were single, had spent time in prison, were unemployed and had at least one parent with an addiction problem or mental illness in comparison to non-abusers (n = 85). They had started using heroin and cocaine earlier and currently abused more cocaine, heroin and cannabis. They had significantly more psychopathology and negative mood. They had significantly more HCV and reported more HIV/HCV risk-taking behavior. IMPLICATIONS: We suggest that this group of patients is in need of more intensive pharmacological and psychological treatment. PMID- 10790906 TI - The 12-month prevalence of substance use and ICD-10 substance use disorders in Australian adults: findings from the National Survey of Mental Health and Well Being. AB - AIMS: To present the prevalence of substance use and ICD-10 substance use disorders in the adult Australian population using data from the National Survey of Mental Health and Well-Being (NSMHWB). DESIGN: A cross-sectional survey assessing substance use and ICD substance use disorders (harmful use and dependence). SETTING AND PARTICIPANTS: A household survey of a nationally representative sample of 10,641 Australian adults (aged 18 years or older). MEASUREMENTS: Trained survey interviewers administered a structured interview based on the Composite International Diagnostic Interview (CIDI). FINDINGS: In the past 12 months 6.5% of the sample had an ICD-10 alcohol use disorder (95% CI: 6.2, 6.9), and 2.2% had another drug use disorder (95% CI: 2.0, 2.4). More males than females had substance use disorders: 9.5% of males (95% CI: 8.5, 10.5) and 3.6% of females (95% CI: 3.2, 4.0) met criteria for an alcohol use disorder, and 3.2% of males (95% CI: 2.8, 3.6) and 1.3% of females (95% CI: 0.9, 1.7) met criteria for another drug use disorder within the past 12 months. The prevalence of substance use disorders decreased with increasing age: 10.5% of respondents aged 18-34 years met criteria for an alcohol use disorder and 4.8% met criteria for a drug use disorder. The rates of these disorders among those aged 55 years or older were 1.8% and 0.1%, respectively. Substance use disorders were more prevalent among the unemployed, those who had never married and those who were Australian-born. CONCLUSIONS: The prevalence of substance use disorders in the Australian population is comparable to that in other English-speaking countries. PMID- 10790907 TI - Exploring the dose-response relationship between alcohol consumption and the risk of several alcohol-related conditions: a meta-analysis. AB - OBJECTIVE: To compare the strength of the evidence provided by the epidemiological literature on the association between alcohol consumption and the risk of six cancers (oral cavity, oesophagus, colorectum, liver, larynx, breast), hypertension, cerebrovascular diseases, gastric and duodenal ulcer, liver cirrhosis and other chronic liver diseases, pancreatitis and injures and adverse effects. METHODS: A search of the epidemiological literature from 1966 to 1998 was performed by several bibliographic databases. Meta-regression models were fitted considering fixed and random models and linear and non-linear effects of alcohol intake on the risk of each condition. The effects of some characteristics of the studies including an index of their quality were considered as putative sources of heterogeneity of the estimates. Publication bias was also investigated by asymmetry of funnel plots. RESULTS: Of the 397 initially reviewed studies, 200 were selected for meta-analysis. Since qualitative characteristics of the studies were often significant sources of heterogeneity among them, the estimates of the pooled dose-response slopes were based only on the 123 studies with higher quality score and/or reporting adjusted estimates of relative risks. Higher alcohol-related risks were found for liver cirrhosis, neoplasms of the upper respiratory and digestive tracts, haemorrhagic stroke and injuries and adverse effects. Weaker but significant associations were found for colorectum, liver and breast cancers, essential hypertension and chronic pancreatitis. For all these conditions, low intakes, corresponding to daily consumption of two drinks or two glasses of wine (25 g/day), have shown significant risks. Ischaemic stroke and gastric and duodenal ulcer seem independent of alcohol intake. The area in which the study was performed, the study's design and the outcome variable differently affected the slopes. CONCLUSIONS: The small number of sufficiently reliable studies, the strong indications of heterogeneity across them and the suspicion of publication bias suggest that there is a great need for well-conducted epidemiological studies performed in several countries, to examine the dose response relationship between alcohol intake and the risk of several alcohol related conditions, as well as the role of drinking pattern in determining the risk. PMID- 10790908 TI - Misuse of diphenhydramine soft gel capsules (Sleepia): a cautionary tale from Glasgow. PMID- 10790910 TI - A man with heart failure refuses to be admitted. PMID- 10790909 TI - When is a DEXA scan indicated? PMID- 10790911 TI - Key developments in rheumatology. PMID- 10790912 TI - The patient with osteoarthritis. PMID- 10790913 TI - Managing the painful shoulder. PMID- 10790914 TI - Controlling hyperuricaemia in general practice. PMID- 10790916 TI - A century of change. PMID- 10790915 TI - The challenge of paper 2. PMID- 10790917 TI - General practice into the next century. PMID- 10790918 TI - The relationship between patient and doctor at the turn of the new century leaves a lot to be desired. PMID- 10790919 TI - Improved communication and higher clinical quality dominate the future of the medical profession. PMID- 10790921 TI - Progress in the management of COPD. PMID- 10790920 TI - Key developments in respiratory medicine. PMID- 10790922 TI - Education for asthma self-management. PMID- 10790924 TI - Tackling the oral exam. PMID- 10790923 TI - Helping smokers to stop: an evidence-based approach. PMID- 10790925 TI - The need to build community autonomy in public health. PMID- 10790926 TI - Public health ethics and the limits of evidence. PMID- 10790927 TI - What value is public health? PMID- 10790928 TI - Public health: just processes? PMID- 10790929 TI - The values of public health: hopefully not a final word... PMID- 10790930 TI - A thorough going over: evidence for health assessments for older persons. AB - OBJECTIVE: To review published randomised controlled trials of health assessments for older people; consider the effects of assessments in maintaining health and quality of life for older people; and identify those factors associated with more successful health assessment programs. METHODS: A systematic literature search and methodological review of published studies of health assessments for people aged 65 years and over, living in the community. RESULTS: Twenty-one trials were identified. They were widely heterogeneous in terms of methodological quality, assessment content and outcome variables. While the studies' results are inconsistent, the majority of the more methodologically sound studies report improvements in health. The studies reporting positive health outcomes were not specifically targeted to particular groups at high need, but were applied to all people in the source population who had reached a set age, usually 75 plus. In the majority of studies reviewed, the assessments were conducted by non-medical personnel (nurse, lay interviewer/volunteer or office staff). CONCLUSIONS: Health assessments have been associated with improved health outcomes for older people. An evidence base for specific components to be included in the assessments is yet to be derived. IMPLICATIONS: In November 1999, new Medicare items to provide for health assessments for persons in Australia aged 75 years and over were introduced. The acceptability, adoption and effectiveness of these items needs careful monitoring. PMID- 10790931 TI - Birthweights and growth of infants in five Aboriginal communities. AB - OBJECTIVES: To improve, by culturally appropriate means, birthweights and growth of children up to three years of age over 14 months in five Aboriginal communities in north-western Australia. METHODS: Frequent individual nutritional assessment of infants and children with counselling of mothers and carers and of pregnant women and the introduction of the Aboriginal-controlled Strong Women, Strong Babies, Strong Culture maternal support program. Outcomes compared with those in the same communities for the five years preceding intervention. RESULTS: By international standards, pre-intervention birthweights of full-term infants (37-42 weeks) were only moderately depressed and recovered to exceed standard weight-for-age within two weeks of birth. Growth of full-term infants slowed abruptly after six months. Prevalence and duration of breastfeeding were very high. Prevailing low average birthweight was chiefly attributable to a prevalence of pre-term birth approaching 20%. Intervention was not accompanied by any change in full-term birth weight but was associated with increased weight gain after six months. From 12 to 36 months growth rose by 30 g per month (p = 0.001). Average birthweights of pre-term infants were < 2,500 g and average weight-for-age did not improve during intervention. CONCLUSIONS: Both low birthweight and a disproportionate part of intransigent failure to grow by Aboriginal infants were associated with pre-term birth. Depressed average growth of full-term infants appeared to respond to nutritional counselling accompanied by a community support program. IMPLICATIONS: Investigation of the causes of the exceptionally high rate of Aboriginal pre-term birth in the region is urgently required. PMID- 10790932 TI - Effectiveness of a community-directed 'healthy lifestyle' program in a remote Australian aboriginal community. AB - OBJECTIVE: To assess the sustainability and effectiveness of a community-directed program for primary and secondary prevention of obesity, diabetes and cardiovascular disease in an Aboriginal community in north-west Western Australia. METHOD: Evaluation of health outcomes (body mass index, glucose tolerance, and plasma insulin and triglyceride concentrations) in a cohort of high-risk individuals (n = 49, followed over two years) and cross-sectional community samples (n = 200 at baseline, 185 at two-year and 132 at four-year follow-ups), process (interventions and their implementation) and impact (diet and exercise behaviour). RESULTS: For the high-risk cohort, involvement in diet and/or exercise strategies was associated with protection from increases in plasma glucose and triglycerides seen in a comparison group; however, sustained weight loss was not achieved. At the community level, significant reductions were observed in fasting insulin concentration but no change in prevalence of diabetes, overweight or obesity. Weight gain remained a problem among younger people. Sustainable improvements were observed for dietary intake and level of physical activity. These changes were related to supportive policies implemented by the community council and store management. CONCLUSIONS: Community control and ownership enabled embedding and sustainability of program, in association with social environmental policy changes and long-term improvements in important risk factors for chronic disease. IMPLICATIONS: Developmental initiatives facilitating planning, implementation and ownership of interventions by community members and organisations can be a feasible and effective way to achieve sustainable improvements in health behaviours and selected health outcomes among Aboriginal people. PMID- 10790933 TI - Aboriginal deaths in Western Australia: 1985-89 and 1990-94. AB - OBJECTIVE: To examine death data for Aboriginal and non-Aboriginal persons in Western Australia (WA) in 1985-89 and 1990-94. METHODS: Population estimates were provided by the Health Information Centre of the WA Health Department based on data from the Australian Bureau of Statistics (ABS). Death data came from the WA Registrar-General's Office. Standard methods were used to obtain rates and levels of significance. RESULTS: Main causes of deaths among Aboriginal males in 1990-94 were circulatory conditions, respiratory, injury and poisoning, neoplasms and endocrine diseases; in Aboriginal females they were circulatory, neoplasms, endocrine diseases, respiratory diseases, and injury and poisoning. From 1985-89 to 1990-94, the Aboriginal male all-cause age-standardised death rates fell 3% (ns) while the non-Aboriginal male rate fell 11% (p < 0.05). The Aboriginal female all-cause death rate rose 11% (ns) while the non-Aboriginal rate fell 5% (p < 0.05). The all-cause death rate ratio (Aboriginal:non-Aboriginal) changed from 2.4 to 2.6 (males) and 2.5 to 2.9 (females). There was a major increase in deaths from endocrine diseases among Aborigines and non-Aborigines. This increase was proportionally much greater among Aborigines. In non-Aborigines there was a significant decrease in deaths from circulatory diseases (mainly ischaemic heart disease); this did not occur among Aborigines. CONCLUSIONS: Over the study period, Aboriginal health standards, as reflected by death rates, apparently worsened relative to non-Aboriginal standards. IMPLICATIONS: Better health promotion, disease prevention and disease care are required to help achieve acceptable health standards among Aboriginal peoples. PMID- 10790934 TI - Comparison of trends in method-specific suicide rates in Australia and England & Wales, 1968-97. AB - OBJECTIVE: To compare secular trends in method-specific suicide rates among young people in Australia and England & Wales between 1968 and 1997. METHODS: Australian data were obtained from the Australian Bureau of Statistics, and for England & Wales from the Office for National Statistics. Overall and method specific suicide rates for 15-34 year old males and females were calculated using ICD codes E950-9 and E980-9 except E988.8. RESULTS: In both settings, suicide rates have almost doubled in young males over the past 30 years (from 16.8 to 32.9 per 100,000 in Australia and from 10.1 to 19.0 in England & Wales). Overall rates have changed little in young females. In both sexes and in both settings there have been substantial increases in suicide by hanging (5-7 fold increase in Australia and four-fold increase in England & Wales). There have also been smaller increases in gassing in the 1980s and '90s. In females, the impact of these increases on overall rates has been offset by a decline in drug overdose, the most common method in females. CONCLUSIONS: Rates of male suicide have increased substantially in both settings in recent years, and hanging has become an increasingly common method of suicide. The similarity in observed trends in both settings supports the view that such changes may have common causes. Research should focus on understanding why hanging has increased in popularity and what measures may be taken to diminish it. PMID- 10790935 TI - Injury hospitalisation rates in Victoria, 1987-97: trends, age and gender patterns. AB - OBJECTIVE: To estimate the injury hospitalisation trends and the age and gender patterns in Victoria between 1987/88 and 1996/97. METHOD: The Victorian Injury Surveillance System maintains a collection of injury hospitalisation records in Victoria. We used loglinear models for rates to estimate the temporal trends and patterns by age group and gender of all-injury and cause-specific injury hospitalisation rates. RESULTS: The all-injury hospitalisation rate appeared stable until 1992/93, after which it increased substantially (1993/94 was 1.13 times that of 1987/88). While motor vehicle crash and fire/burn/scald injury hospitalisation rates decreased substantially, suicide attempt and self-inflicted injury hospitalisation rates more than doubled. The all-injury hospitalisation rate was highest for persons 75 years of age and older. Females had a lower all injury hospitalisation rate than males up to 60-64 years of age and a higher rate thereafter. The cause-specific injury hospitalisation rates showed various age and gender patterns. CONCLUSION: Substantial increases in the all-injury hospitalisation rate were observed after the introduction of the casemix funding policy to public hospitals. There were remarkable variations in temporal trend and age and gender patterns of injury hospitalisation rates by cause of injury. IMPLICATIONS: Hospitalised injuries generally are serious and incur high cost. Certain injury causes and age and gender groups should receive particular attention for prevention efforts. The use of injury surveillance based on hospitalisation records should be improved and further expanded. PMID- 10790936 TI - Perceived needs of women diagnosed with breast cancer: rural versus urban location. AB - OBJECTIVE: To assess the prevalence and predictors of perceived unmet needs among women diagnosed with breast cancer, with a particular focus on the impact of location (rural/urban). METHOD: A random sample of women in NSW diagnosed with breast cancer was surveyed during 1995-96. The Breast Cancer Patients' Needs Questionnaire (BR-CPNQ) explores patients' perceived needs across five domains: psychological, health information, physical/daily living, patient care/support and interpersonal communication. Items assessing breast cancer specific needs and access to services and resources were included. Of 235 eligible rural women, 134 (57%) consented to participate and 129 (55%) returned completed surveys. Of 196 eligible urban women, 102 (52%) consented to participate and 100 (51%) returned completed surveys. RESULTS: For 12 of the 52 items, at least one-third of the sample reported ever having a moderate/high need for help. Of the 15 highest moderate or high unmet needs, 10 related to health information and three to psychological needs. There were few differences in the prevalence of needs reported by rural and urban women. Only in the physical/daily living domain were rural women more likely than urban women to report some need for help (OR = 2.59, 95% CI 1.21-5.52). CONCLUSIONS: Unmet needs, particularly in the information and psycho-social domains, are reported by both rural and urban women with breast cancer, with sub-groups experiencing different types of needs. IMPLICATIONS: There is a need to develop and trial targeted and innovative strategies to meet the health information and psychological needs of women with breast cancer, and the physical/daily living needs of rural women diagnosed with breast cancer. PMID- 10790937 TI - Acute effects of bushfires on peak expiratory flow rates in children with wheeze: a time series analysis. AB - OBJECTIVES: To determine the effects of the January 1994 Sydney bushfire on evening peak expiratory flow rates (PEFR) in children with wheeze. METHODS: Children with a history of wheeze were enrolled in the longitudinal study and completed a daily asthma diary. We obtained daily air pollution, meteorological, pollen and alternaria data. We then used generalised estimating equation techniques to determine associations between the bushfire period and particulate matter less than 10 microns (PM10) and PEFR. RESULTS: The maximum daily PM10 level peaked at 210 ug/m3, which was nearly seven times the usual PM10 level for the rest of January and February 1994. There was no significant association between mean PM10 and PEFR (beta-coefficient = -0.009, p = 0.86). Children without bronchial hyper-reactivity had a significant negative association between PEFR and PM10 (beta-coefficient = -0.1029, p = 0.03). The bushfire period was not significant in any of the models. CONCLUSIONS: We did not find an association between the bushfire period or PM10 and evening PEFR, although in a subgroup of children without bronchial hyper-reactivity, a significant negative association was present between PM10 and evening PEFR. IMPLICATIONS: We conclude that the high levels of particulate pollution caused by the Sydney bushfires did not lead to any clinically significant reductions in PEFR in symptomatic children. Our results have implications for community risk communication during future bushfires. PMID- 10790938 TI - Sun exposure and primary prevention of skin cancer for infants and young children during autumn/winter. AB - OBJECTIVE AND METHOD: Excessive sun exposure in childhood has been associated with the development of skin cancer, and appropriate levels of sun protection during the early years of a child's life can significantly reduce the risks. A survey of 133 mothers was undertaken in south-east Queensland to examine the levels of sun exposure and skin protection of infants and young children. RESULTS: Among this sample of 133 mothers, use of appropriate skin protection was relatively high for themselves and their children. However, even by six months of age, a third had been sunburnt and 15% had experienced painful sunburn. By three years of age, 82% had been sunburnt and one-third had experienced painful sunburn, though mothers' knowledge levels of sun safety issues were very high. The predictors associated with primary prevention varied across the type of prevention behaviour, but indicated that interventions should focus on susceptibility to sunburn and history of sunburn. Similar programs could be applicable across a broad range of socio-demographic groups, but require attention to mother's country of birth as a factor that significantly influences their use of protection for their young children. CONCLUSIONS: Ongoing public education that targets specific groups and settings may contribute to adoption of appropriate sun protective behaviours for young children. IMPLICATIONS: Results of the study provide some of the needed baseline data to assist future skin cancer prevention campaigns for young children and infants. A similar study of levels of sun exposure and skin protection for children during summer is under way. PMID- 10790939 TI - Patterns of alcohol consumption in young Australian women: socio-demographic factors, health-related behaviours and physical health. AB - OBJECTIVE: To determine which socio-demographic factors, health-related behaviours and physical health conditions are associated with non-drinking, binge drinking and hazardous/harmful drinking in young Australian women. METHODS: Cross sectional data were obtained from the baseline survey of 14,762 young women (aged 18-23 years) enrolled in the Women's Health Australia study in 1996. Associations between a range of drinking patterns and socio-demographic factors, health related behaviours and health conditions were examined. RESULTS: Half the women were 'low intake' drinkers, a third 'rarely drank' and 9% were non-drinkers; however, 70% reported binge drinking with one-quarter of the binge drinkers doing so at least weekly. Non-drinkers were more likely than drinkers to be married, pregnant, non-smokers, born in non-English speaking countries, to live in the Northern Territory, and to have lower levels of education, employment, and private health insurance. 'Low intake/binge weekly' drinkers (12%) and 'hazardous/harmful' drinkers (5%) were more likely than 'low risk' drinkers to be unmarried; to live in shared accommodation, alone or with their parents; to live in rural or remote areas; to have ever had any sexually transmitted infection; to be current smokers or ex-smokers and to have used unhealthy weight-control practices. CONCLUSIONS: The results confirm findings from other countries about the importance of social conditions as determinants of alcohol consumption by young women. IMPLICATIONS: Health promotion to reduce young women's alcohol consumption needs to be carefully targeted to take account of their demographies, living environments and beliefs. PMID- 10790940 TI - A case study of health goals in New Zealand. AB - OBJECTIVE: This paper outlines the New Zealand experience in using health goals and examines its strengths and weaknesses from an 'insiders's' perspective. METHOD: This paper reports on a review of the New Zealand health goals framework conducted in 1996-97. The review centred on a discussion paper, written submissions on it, and consultation meetings with the public, the public health sector and relevant government agencies. RESULTS: It is argued that the framework usefully shaped public health activity in New Zealand and should be retained with a focus on strengthening public health action. Health goals have been developed in New Zealand at a time of considerable change in the health sector. Although this change has been disruptive, it has also provided benefits such as the emergence of new providers. The strengths of the New Zealand framework have included: its inclusiveness, the consultation that occurred in developing it, and the monitoring and reporting system. Ongoing challenges, such as reorienting the health sector and developing a formal intersectoral strategy, are also identified. CONCLUSION: The paper concludes that the current health goals framework has the potential to frame future public health action in New Zealand, but that the increasing mainstreaming of the public health function poses some risk. IMPLICATIONS: The insight provided by the New Zealand case on the implementation of a health goals framework may assist public health planners in other jurisdictions. PMID- 10790941 TI - Public health surveillance of hepatitis C: can it identify incident cases? AB - OBJECTIVES: To evaluate a follow-up system to identify incident cases among individuals notified with the hepatitis C virus (HCV). METHOD: A cross-sectional survey of medical practitioners treating individuals notified to the NSW Health Department as having HCV between August 1996 and August 1997 was conducted. RESULTS: Five hundred and fifty-four new notifications were received during the study period (70.7 per 100,000 people). Ninety-six per cent of notifications were followed up with 54 individuals (9.7%) identified as incident cases. Incident cases were significantly younger than prevalent cases (median age 30 vs. 39, p < 0.001) with drug and alcohol notifications being more likely to be incident cases. CONCLUSION: HCV transmission is continuing at relatively high levels with incident cases being significantly younger than prevalent cases. IMPLICATIONS: An efficient notification follow-up strategy that identifies incident cases could be routinely used to assess the effectiveness of population-based initiatives aimed at reducing HCV transmission. PMID- 10790942 TI - The Australian National Death Index: an assessment of accuracy. AB - OBJECTIVE: The Australian National Death Index (NDI) provides a comprehensive and accessible source of mortality information for epidemiological research. Use of the index requires a probabilistic matching process that inevitably results in some inaccuracy. In this paper, accuracy is assessed. METHODS: Results of a matching process against the NDI performed by the Australian Institute of Health and Welfare in Canberra were compared with information provided by the Medical Device Outcomes study cohort and their families (n = 2,990). Indices of accuracy for the NDI were calculated. RESULTS: For this particular study, the NDI has sensitivity 88.8% (84.9-92.8) and specificity 98.2% (97.4-98.7). CONCLUSIONS AND IMPLICATIONS: The relatively low sensitivity is of some concern to those using the NDI for health outcomes research. The importance of such a national database is evident; however, to improve accuracy the introduction of a national unique patient identifier is necessary. PMID- 10790943 TI - Parental use of a paediatric emergency department as an ambulatory care service. AB - OBJECTIVE: This qualitative study explored the parental attitudes, perceptions and beliefs that play a role in the use of a tertiary paediatric emergency department (PED) when a child has a non-urgent illness. METHOD: Semi-structured, in-depth interviews of 25 parents of children with non-urgent illnesses were conducted in the waiting room of a tertiary PED in Western Sydney in 1998. Inductive analysis was used to identify dominant themes. RESULTS: Parents used their own system of triage to choose the appropriate service for their sick child. The perceived expertise of the tertiary PED, access and parental expectations all appeared to be major factors in parental use of a PED. CONCLUSIONS: The parental choice to attend a PED is a dynamic, complex and unique process and the parental views that underpin this process often diverge from those of health professionals about the most 'appropriate' use of a PED. IMPLICATIONS: A clearer understanding by health professionals of the factors influencing parental choice will promote more effective collaboration with parents and ultimately assist in the decision on the best management option for sick children. PMID- 10790945 TI - Reconciling pedagogy and health sciences to promote indigenous health. AB - OBJECTIVES: To increase knowledge and skills regarding Indigenous learning styles. To raise awareness within the tertiary education sector that Aboriginal students learn differently and that Indigenous cultures and pedagogy have validity and strength. To examine pedagogical strategies that assist both tertiary students capacity for learning and university lecturers' delivery and evaluation of teaching and learning strategies. METHODS: A qualitative, ethnographic framework using personal observations, field and classroom experience, interviews and review of literature in the fields of education, public health and Indigenous cultural perspectives. RESULTS: Aboriginal people are the receivers of services and programs that will be delivered, in the majority of cases, by university-educated, non-Aboriginal, professional health care providers. Indigenous students face specific challenges in obtaining an effective education for working in the Aboriginal and wider community in the field of public health; the challenges relate to culture, health paradigms and community. CONCLUSION: Lecturers in health and human science courses for Aboriginal students need to both examine and appreciate the cultural constraints on learning faced by their students within the context of mainstream curriculum, and to build on the large pool of knowledge and learning styles that Aboriginal society bequeaths to Aboriginal students. IMPLICATIONS: Academics can apply the cultural differences and knowledge base of the Indigenous community as a force to promote health through learning. PMID- 10790944 TI - The Child Health Questionnaire in Australia: reliability, validity and population means. AB - OBJECTIVE: To provide reliability, validity and population means for the Australian Authorized Adaptation of the parent-report Child Health Questionnaire (CHQ). METHOD: We surveyed a representative sample of Australian parents of school-aged children (5-18 years) in Victoria between July and December 1997, using a school-based cluster sample design stratified by educational sector and age. RESULTS: Some 5,414 parents responded (72%). Good psychometric performance was observed for the CHQ in Australia. Population means demonstrated differences in health on domains of functioning and well-being by age and gender. This population-derived sample demonstrated high ceiling values on Physical Functioning and Social Role scales. IMPLICATIONS: The CHQ appears to be a reliable and valid measure of child and adolescent functional health and well being for the Australian population. Child health outcomes of children and adolescents with particular conditions or within population subgroups can be compared with these age and gender benchmarks. Appropriate uses for the CHQ may be to discriminate between children who are generally healthy and children with health problems, or in population surveys partnered with measures that extend the range of physical functioning and social functioning. PMID- 10790946 TI - What can a trial contribute to the debate about supervised injecting rooms? AB - Outlines the areas for evaluation of supervised injecting rooms, the limitations of any evaluation and how the evaluation results can be buttressed against misuse. Argues that forceful advocacy for the role of evaluation in political decision-making is needed. PMID- 10790947 TI - TV towers and childhood leukaemia (continued) PMID- 10790948 TI - Co-ordinated Care Trials and change in rural health systems. PMID- 10790949 TI - I.Q.H., information and quality healthcare: improving the care of patients with acute myocardial infarction and congestive heart failure. AB - Acute myocardial infarction and congestive heart failure are prevalent conditions in Mississippi associated with significant mortality and morbidity. It is critically important that physicians and hospital staffs work together to implement system and process changes that will facilitate the appropriate, timely evaluation and use of indicated therapies in both AMI and CHF patients. Change, while never easy, can be beneficial when properly motivated, designed and implemented. Physicians can and do make a difference not only in the quality of care given but also in the quality of life lived by their patients. I.Q.H. is encouraging all Mississippi physicians and healthcare facilities to examine their treatment processes for AMI and CHF. PMID- 10790950 TI - The ABC's of AMI. PMID- 10790951 TI - Overview of current chronic heart failure therapy. PMID- 10790952 TI - Coronary care 101. PMID- 10790954 TI - Thank you very much PMID- 10790953 TI - Mississippi State Hospital opens a museum: a history of the treatment of mental illness in Mississippi. PMID- 10790955 TI - [Gastric lesions induced by non-steroidal antiinflammatory drugs (part I)]. PMID- 10790956 TI - [Gout crisis]. PMID- 10790957 TI - [Music therapy: history and perspectives]. PMID- 10790958 TI - [Nitric oxide synthesis in patients with myocardial infarction]. AB - The findings of the author demonstrate that the system of nitric oxide (NO) generation and metabolism is an element of multicomponent response of the organism to myocardial infarction (MI). This response consists in MI patients' systemic ability and, in particular, their peripheral blood mononuclears' ability to produce NO as well as the absence of iNOS activation in peripheral blood of MI patients. It is important for a practitioner to understand that a fall in the urine and plasma concentration of NO metabolism end product reflects low activity of NO generation while NO is a powerful regulatory factor in the cardiovascular system. Thus, low levels of NO and its metabolites in the urine and plasma of MI patients indicate depletion of compensatory coronarodilatating potential and, eventually, poor prognosis. Relevant measurements will provide additional parameters in assessment of body reserves in MI patients and in MI prognosis within the first hours of its onset. PMID- 10790959 TI - [Characteristics of microcirculation and vascular responsiveness in elderly patients with hypertension and ischemic heart disease]. AB - Microcirculation and vascular responsiveness were studied in 52 patients with arterial hypertension and ischemic heart disease versus 48 healthy elderly persons. The patients were found to have defects of the end blood flow in all links of microcirculation, longer and more severe vasoconstriction of conjunctival and skin vessels in response to norepinephrine and cold stimulation tests. PMID- 10790960 TI - [Risk factors of ischemic heart disease and lipid metabolism in cardiorespiratory diseases]. AB - Clinicobiochemical parameters and factors of risk of ischemic heart disease in patients with cardiorespiratory diseases (CRD) were examined for correlations. The risk factors, impairment of lipid spectrum and antioxidant defense were assessed in patients aged 27 to 70 years with CRD. The patients were divided into three groups: patients with chronic nonspecific pulmonary diseases (CNSPD), ischemic heart disease patients (IHD), those with CNSPD plus IHD. The results suggest that in CNSPD patients such risk factors as smoking, low physical activity, stress, arterial hypertension, hereditary predisposition to IHD occurred in more than 50% of cases. Compared to IHD patients, these patients' dyslipoproteinemia was characterized by greater frequency of hypercholesterolemia and less frequency of hypoalphacholesterolemia. Combination of atherosclerosis with CNSPD aggravates the course of vascular disorders. PMID- 10790961 TI - [Program of examination for patients with isolated mitral valve defects to evaluate the degree of pulmonary hypertension]. AB - 98 patients with isolated rheumatic mitral valve defects of the heart entered the study. Their central hemodynamics was studied intraoperatively before surgical correction of the defect. The results were compared to those provided by conventional noninvasive methods: chest x-ray, ECG, echo-CG, external respiration function test. Correlation analysis using computer intellect PolyAnalyst v.1.01R helped to design a program and practical nomogram which can quantitatively determine the degree of pulmonary hypertension on the basis of digital values of Rv1 wave amplitude (ECG) and right ventricular cavity size (echoCG). PMID- 10790962 TI - [Mental status of patients with iodine-deficiency goiter]. AB - Personality and intellectual characteristics were examined in 76 patients with iodine-deficiency goiter (IDG). Subclinical hypothyroidism forming as early as IDG of the 1st degree impairs abstractive and integral thinking, psychological maladaptation to the disease, high anxiety, neurotism, diminished intellectual performance, deterioration of memory, attention, predominance of thinking of concrete-action type. To prevent intellectual disturbances, IDG patients should receive replacement therapy with adequate doses of thyroid hormones preparations. PMID- 10790964 TI - [Significance of chemiluminescent tests in assessing efficacy of ulcer treatment with microwave resonance]. AB - To study prognostic value of chemiluminescent (CL) test of duodenal ulcer treatment efficacy with microwave resonance therapy (MWRT), 39 duodenal ulcer patients were divided into two groups differing only by the size of ulcer defect. In 23 patients of the first group scarring occurred after 10 MWRT procedures. In 16 patients of the second group scarring of the ulcer occurred after 10-20 procedures. By the level of lipid hydroperoxides and antioxidant activity it was found that rapid healing of ulcer is accompanied with a positive change in the total antioxidant activity whereas slow healing in MWRT occurs in reduced total antioxidant potential and seems to depend on the size of ulcer defect. PMID- 10790963 TI - [Biochemical markers of inflammation and bronchial obstruction]. AB - Bronchoalveolar lavage (BAL) was studied in 139 patients with chronic obstructive pulmonary diseases (COPD). 90 of them had chronic bronchitis, 35--bronchial asthma. Overall metabolites of nitric oxide, histamine, serotonin, malonic dialdehyde were measured. Correlations were studied between concentrations of biologically active compounds and lung ventilation in exacerbation and remission. It was found that levels of indolalkylamines change in one direction both in inflammation and bronchial obstruction. Lipid peroxidation in BAL is closely connected with inflammation in both the diseases and is not related with disorders in lung ventilation. Nitric oxide metabolites differed: gas secretion in bronchial asthma exacerbation is higher than in chronic obstructive bronchitis. In its remission, nitric oxide activity is low, while in bronchial asthma it persisted higher than in healthy subjects. PMID- 10790966 TI - [Josamycin combination with ranitidine in therapy of gastroduodenal ulcer: clinical efficiency]. AB - Efficacy of a novel eradication treatment with ranitidine, trichopol and jozamycin was compared to that of the standard three-component therapy and monotherapy with ranitidine in 76 patients with exacerbation of gastric and duodenal ulcer. Therapeutic monitoring of ranitidine was performed. In relation to healing of ulcer defect and eradication of Helicobacter pylori, jozamycin is not less effective and more safe than standard three-component therapy. Interaction of the scheme components increases plasma concentration of ranitidine especially in patients with concomitant hepatic and renal diseases. PMID- 10790965 TI - [Effects of angiotensin-converting enzyme (ACE) inhibitors on water-salt homeostasis in hypertensive patients living in Far North]. AB - The aim of the study was evaluation of ACE inhibitors (captopril and ramipril) effect on water-salt homeostasis in the treatment of patients with arterial hypertension (AH) living in the Far North of Russia. 100 male patients with mild and moderate AH were examined 2 weeks, 3 and 6 months after administration of captopril or ramipril. The drugs are shown to correct water-salt metabolism. This is explained by better renal function due to speeding up glomerular filtration and increased sodium excretion with urine, and by activity of humoral mechanisms (inhibited activity of plasma renin, low plasma concentration of aldosterone and its 24-h excretion). Comparison of captopril versus ramipril demonstrates advantages of prolonged ramipril in respect to regulation of water salt metabolism in the treatment of essential hypertension in the Far North. PMID- 10790967 TI - [Immunostimulating activity of eubiotic bioflor in intestinal dysbacteriosis of various origin]. AB - The data are reported on novel eubiotic biofor's effects on immune status of 213 patients with intestinal dysbacteriosis (39 surgical gastroenterological patients, 43 patients with diarrheal infections, 50 AIDS patients, 81 cancer patients exposed to radiation and chemotherapy). It was found that bioflor acts as an antidysbacteriosis drug and as an immunostimulator. It stimulates T-cell immunity, phagocytosis, B-cell immunity. Immunostimulating effect of bioflor depends on dysbacteriosis degree and is maximal at its first stages. PMID- 10790968 TI - [Gastrointestinal diseases in patients with bronchial asthma]. PMID- 10790969 TI - [Optimization of rehabilitation system of ischemic heart disease patients after coronary artery bypass surgery]. PMID- 10790970 TI - [Helicobacter pylori in chronic gastroduodenal ulcer]. PMID- 10790971 TI - Using ROC curves to test models of recognition memory: the relationship between presentation duration and slope. AB - Many theories of memory predict that the slope of the z-transformed receiver operating characteristic (ROC) curve should decrease with increases in the memory strength of old items. While several prior studies have failed to demonstrate this effect, the results of two experiments demonstrate that increasing presentation duration can reduce the slope of the z-transformed ROC curve. These results raise questions about the generality of prior results and of dual-process theories designed to accommodate those results. We close by emphasizing that determining the experimental circumstances that affect whether or not the slope of the z-transformed ROC curve decreases will provide important constraints on theories of recognition memory. PMID- 10790972 TI - Recollection-based recognition eliminates the revelation effect in memory. AB - The revelation effect refers to the tendency to call an item on a recognition test old if it is preceded by a cognitive task that involves the processing of a similar stimulus (Watkins & Peynircioglu, 1990). It has been proposed that the revelation effect occurs because of an increase in the familiarity of the test items in the revelation condition (Luo, 1993; Westerman & Greene, 1998). In the present experiments, the revelation effect was investigated in recognition tasks that were not based solely on the familiarity of the test items but, also, on a recall-like retrieval process. A revelation effect was not found on an associative-recognition task or on a plurality recognition task. The results of this study show that the revelation effect does not occur when the contribution of familiarity to recognition decisions is reduced by factors that encourage the recollection of the study episode. PMID- 10790973 TI - The revelation effect for item and associative recognition: familiarity versus recollection. AB - The revelation effect occurs when items on a recognition test are more likely to be judged as being old if they are preceded by a cognitive task that involves the processing of similar types of stimuli. This effect was examined for item (single word) and associative (word-pair) recognition. We found, in Experiments 1 and 2, a revelation effect for item, but not for associative recognition under normal study conditions. A revelation effect for both item and associative recognition was observed in Experiments 3 and 4 when study time was extremely brief, thus limiting the encoding of information that would support recall or recollection. In Experiment 5, we demonstrated that the revelation effect for item recognition is eliminated when item recognition decisions are made in the context of a study item. The results show that the revelation task influenced recognition decisions based on familiarity, but not decisions that involved recall or recollection. PMID- 10790974 TI - Visual encoding of patterns is subject to dual-task interference. AB - Two stimuli were presented at varying stimulus onset asynchronies (SOAs), with each stimulus associated with a distinct task. The first stimulus was a tone at one of either two or three frequencies. In two conditions, the task associated with a tone was either a speeded two-alternative discrimination (2AD), or a speeded three-alternative discrimination (3AD) based on the pitch of the tone. In a third condition, subjects were told to ignore the tone. The second stimulus was a briefly exposed study matrix of red and black squares followed by a mask. After a fixed delay, the mask was replaced by a test matrix that was either the same or different from the study matrix. The task associated with the matrices was to indicate, with no speed pressure, whether the study matrix and the test matrix were the same or different. Results from each speeded AD condition showed that subject's accuracy in the matrix task decreased as the SOA between the tone and the study matrix decreased. This effect was larger for the 3AD tone task than with a 2AD tone task. In addition, within each speeded AD condition, longer RTs in the tone task were associated with lower accuracy in the matrix task. None of these effects was evident when the subjects were told to ignore the tone. These results suggest that encoding visual information can be subject to significant capacity limitations imposed by cross-modal multitasking. PMID- 10790975 TI - Testing instance models of face repetition priming. AB - Two experiments examining repetition priming in face recognition are reported. They employed eight rather than the more usual two presentation trials so that the prediction made by Logan's (1988) instance model of power function speedup of response time (RT) distributions could be examined. In Experiment 1, we presented the same photograph on each trial; in Experiment 2, we presented photographs of varying poses. Both experiments showed repetition priming effects for familiar and unfamiliar faces, power function speedup for both the mean and the standard deviation of RT and the power function speedup of the quanties of the RT distributions. We argue that our findings are consistent with the predictions made by the instance model and provide an explanatory challenge for alternative theoretical approaches. PMID- 10790977 TI - Do film cuts facilitate the perceptual and cognitive organization of activity sequences? AB - Film depictions of activities possess two kinds of structures--namely, the structural features of the depicted activities themselves and a formal structure defined by film cuts. The former structure is used by everyday observers for perceptually and cognitively unitizing the continuous flow of events into comprehensible entities. It seems conceivable that cuts can serve a similar unitizing purpose for film viewers. For each of two different activity sequences, two film versions were produced. Throughout each film version, cuts were placed either at breakpoints or at nonbreakpoints. In a 2 x 2 (activity sequence x film version) factorial design, 40 subjects segmented the film during viewing and recalled the film content after viewing in a detailed protocol. Segmentation behavior depended primarily on the occurrence of breakpoints and was largely unaffected by the occurrence of cuts. Cuts accompanying a breakpoint lead to more detailed recall protocols for these sections of the film. PMID- 10790976 TI - On the interaction between linguistic and pictorial systems in the absence of semantic mediation: evidence from a priming paradigm. AB - Use of the letter search task in the context of the priming paradigm has proved to be an invaluable tool for the investigation of the strategic control of processes involved in word recognition. In particular, previous findings that letter search on a prime word interferes with the priming of semantically/associatively related targets, but not with the priming of either identical or morphologically related targets, suggests that letter search may selectively interfere with semantic processing, leaving other levels of processing intact. In the present experiments, this investigation was extended by exploring the priming of pictures following letter search of either a same concept word (repetition priming) or a semantically/associatively related word (semantic priming). There was significant repetition priming of picture categorization following both silent reading and letter search of the prime word (Experiments 1 and 2). In contrast, semantic priming of pictures was found only following silent reading of the prime; there was no semantic priming following letter search of the prime (Experiment 2). This pattern of results suggests that focusing attention at the letter level during prime processing selectively attenuates activation of the semantic system by the prime. It does not prevent the spread of activation between the lexical and pictogen levels of representation of a given concept. PMID- 10790978 TI - Priming effects in attentional gating. AB - Three experiments were conducted to determine the effects of amount of prior target information (Experiment 1) and semantic priming (Experiment 2) in an attentional gating task. The goal was to determine some causes of the processing deficits commonly observed in perceiving successive visual stimuli. Items in a rapid serial visual presentation (RSVP) stream are subject to processing deficits before they are processed to the level of recognition (early selection) and after they have been recognized (late selection). Deficits in the former case presumably are due to an early filter that prevents complete recognition and semantic analysis, whereas deficits in the latter case arise from interference or response competition, producing forgetting among a set of recognized items. The semantic-priming effects found between a cue and a target (Experiment 2) and between two successive targets (Experiment 3) indicate that top-down processes can increase the subjective availability of related items. The results are consistent with the idea that most processing deficits observed in search through an RSVP sequence are due to limited capacity in our ability to form episodic representations of all the items in the sequence. PMID- 10790979 TI - The influence of global discourse on lexical ambiguity resolution. AB - The influence of global discourse on the resolution of lexical ambiguity was examined in a series of naming experiments. Two-sentence passages were constructed to bias either the dominant or the subordinate meaning of a homonym that was embedded in a locally ambiguous sentence. The results provided evidence for the immediate (0-msec interstimulus interval) resolution of lexical ambiguity and were subsequently replicated in Experiment 2, in which an 80-msec stimulus onset asynchrony exposure duration was employed for the homonyms. Strong dominant and subordinate biased discourse contexts activated only the contextually appropriate sense of a homonym. In Experiment 3, each sentence of the discourse was presented in isolation. The pattern of activation obtained in Experiments 1 and 2 was found to be contingent on the integration of the two sentences to construct an overall global discourse representation of the text. The results support a context-sensitive model of lexical ambiguity resolution. PMID- 10790980 TI - The basis of transfer in artificial grammar learning. AB - In two experiments, we examined the extent to which knowledge of sequential dependencies and/or patterns of repeating elements is used during transfer in artificial grammar learning. According to one view of transfer, learners abstract the grammar's sequential dependencies and then learn a mapping to new vocabulary at test (Dienes, Altmann, & Gao, 1999). Elements that are repeated have no special status on this view, and so a logical prediction is that learners should transfer as well after exposure to a grammar without repetitions as after exposure to a grammar with them. On another view, repetition structure is the very basis of transfer (Brooks & Vokey, 1991; Mathews & Roussel, 1997). Learners were trained on grammars with or without repeating elements to test these competing views. Learners demonstrated considerable knowledge of sequential dependencies in their training vocabulary but did not use such knowledge to transfer to a new vocabulary. Transfer only occurred in the presence of repetition structure, demonstrating this to be the basis of transfer. PMID- 10790981 TI - The temporality effect in counterfactual thinking about what might have been. AB - When people think about what might have been, they undo an outcome by changing events in regular ways. Suppose two contestants could win 1,000 Pounds if they picked the same color card; the first picks black, the second red, and they lose. The temporality effect refers to the tendency to think they would have won if the second player had picked black. Individuals also think that the second player will experience more guilt and be blamed more by the first. We report the results of five experiments that examine the nature of this effect. The first three experiments examine the temporality effect in scenarios in which the game is stopped after the first contestant's card selection because of a technical hitch, and then is restarted. When the first player picks a different card, the temporality effect is eliminated, for scenarios based on implicit and explicit negation and for good outcomes. When the first player picks the same card, the temporality effect occurs in each of these situations. The second two experiments show that it depends on the order of events in the world, not their descriptive order. It occurs for scenarios without preconceptions about normal descriptive order; it occurs whether the second event is mentioned in second place or first. The results are consistent with the idea that the temporality effect arises because the first event is presupposed and so it is immutable; and the elimination of the temporality effect arises because the availability of a counterfactual alternative to the first event creates an opposing tendency to mutate it. We sketch a putative account of these effects based on characteristics of the mental models people construct when they think counterfactually. PMID- 10790982 TI - Illusions in modal reasoning. AB - According to the mental model theory, models represent what is true, but not what is false. One unexpected consequence is that certain inferences should have compelling, but invalid, conclusions. Three experiments corroborated the occurrence of such illusions in reasoning about possibilities. When problems had the heading "Only one of the premises is true," the participants considered the truth of each premise in turn, but neglected the fact that when one premise is true, the others are false. When two-premise problems had the heading "One of the premises is true and one is false," the participants still neglected the falsity of one of the premises. As predicted, however, the illusions were reduced when reasoners were told to check their conclusions against the constraint that only one of the premises was true. We discuss alternative explanations for illusory inferences and their implications for current theories of reasoning. PMID- 10790983 TI - Knowledge partitioning: context-dependent use of expertise. AB - Although exceptional performance is a defining attribute of expertise, experts sometimes exhibit striking errors and performance limitations. This article reports two experiments in which experts predicted the spread of bush fires, a domain characterized by complex but well-understood physical dynamics. Although accuracy was typically high, large errors were observed when two primary predictor variables were in opposition. In a second study, the experts' behavior- in contrast to that of novices--was additionally shown to depend on problem context. In one context, experts again committed errors, whereas in another, equally domain-relevant context, the correct predictions were made. Critically, when comparing performance across contexts, completely opposing predictions were made under identical physical conditions. We therefore suggest that expertise may comprise separate, and sometimes even mutually exclusive, components of knowledge. PMID- 10790984 TI - The initial segment strategy: a heuristic for route selection. AB - People often choose one route when traveling from point A to point B and a different route when traveling from point B to point A. To explain these route asymmetries, we propose that people rely on a heuristic (the initial segment strategy, or ISS) during route planning. This heuristic involves basing decisions disproportionately on the straightness of the initial segments of the routes. Asymmetries arise because the characteristics that favor selection of a particular route in one direction will usually differ from those that favor selection when traveling in the opposite direction. Results from five experiments supported these claims. In the first three experiments, we found that subjects' decisions were asymmetric and involved a preference for initially straight routes. In Experiment 4, we confirmed that the ISS is a heuristic by demonstrating that people rely on it more when under time pressure. However, people can choose the optimal route when instructed to do so. In Experiment 5, we generalized the findings by having subjects select routes on maps of college campuses. Taken together, the results indicate that the ISS can account for asymmetries in route choices on both real and artificial maps. PMID- 10790985 TI - Smallpox: a possible public health threat, again. PMID- 10790986 TI - Toxic epidermal necrolysis secondary to vancomycin. PMID- 10790987 TI - Searching for respite: effective pain management. PMID- 10790988 TI - Steps forward and backward within the fluoroquinolone class. PMID- 10790989 TI - Adaptation to childhood parental loss: the experience of growing up in an "orphanage" and its meaning over time. AB - Intrigued by a newspaper ad in the Argus Leader, placed by The Sioux Falls Children's Home Society in their hope of reuniting adults formerly institutionalized there as children, I wondered what we might learn from the experiences of people having grown up as "orphans." After exploration, I was invited to attend their reunion to facilitate some of the discussion of former residents who were "welcomed home" from various parts of the country. The participants expressed a wide range of emotions as they rediscovered one another- sometimes from childhood memory, sometimes contrasting childhood faces in old photos with the faces of people now present in middle or old age. As the group reminisced, one man told of his calling out the home's upper-story window each evening to his older sister who lived a floor below. He recalled his sadness on the day when there was silence in response, as she had been sent to a foster home. A woman "alumni" sang word for word, with others joining in, an old Christian hymn that they had sung decades before. And, an elderly woman described how the church across the street from the original children's home facility on tenth street, had a large stain glass window depicting the outstretched arms of God. She told how every night, when it was time for her to go to bed, the window would be lit up. Movingly, she related how she would lie in her bed and look through the window of her room, feeling "held" until she fell asleep. Over the many months that followed, men and women who had been institutionalized there throughout childhood entrusted to me their life stories. This article is a summary of their narratives. In general, it seemed that the participants were set apart by the enormity of the poverty of their early years, the enduring quality of their losses, and by a certain tenacity, decency, and striving for restoration. PMID- 10790991 TI - [Cancer therapy and processing information]. AB - The National Cancer Center Japan started to provide free, computer-based cancer information, through a service called the National Cancer Center--Cancer Information Service (NCC-CIS), to patients and their families, physicians and other health professionals via fax (telephone) and the Internet in 1996. NCC-CIS in Japanese is modeled on the Physician Data Query service produced by the National Cancer Institute in the United States and is produced from an independent standpoint adopted to the medical and social environment in Japan. NCC-CIS provides up-to-date cancer treatment information (staging, prognosis and state-of-the-art treatment by stage of disease), supportive care for each type of cancer, an image reference database, and a directory of organizations and lecture meeting reports in Japan in order to facilitate better understanding of cancer among people and support the decision-making process for physicians in order to achieve a reduction in cancer deaths. PMID- 10790990 TI - Treatment of attention-deficit/hyperactivity disorder. AB - OBJECTIVES: To determine (a) the long-term and short-term effectiveness and safety of pharmacological and nonpharmacological interventions for attention deficit/hyperactivity disorder (ADHD) in children and adults and (b) whether combined interventions are more effective than individual interventions. SEARCH STRATEGY: MEDLINE (from 1966), CINAHL (from 1982), HEALTHStar (from 1975), PsycINFO (from 1984), EMBASE (from 1984), and the Cochrane Library searches were completed in November 1997. Reference lists of eligible studies and files of members of the research team and partner organizations were also searched. SELECTION CRITERIA: Studies were selected if they focused on the treatment of ADHD in humans and were published in any language as a full report in peer reviewed journals. Studies including conditions other than ADHD were reported if separate subgroup analyses for patients with ADHD were provided. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data for 41 variables on general characteristics, along with detailed information on interventions, outcomes, and tests. Differences were resolved by consensus or by a third researcher. Studies were not combined quantitatively because the quality of reporting was low and heterogeneity existed across outcome measures and tests. MAIN RESULTS: Seventy-eight studies (77 randomized controlled trials) met the inclusion criteria. Twenty-three studies compared drugs and showed few, if any, differences among methylphenidate (MPH), dextroamphetamine (DEX), and pemoline; studies comparing stimulants with tricyclic antidepressants (2) were inconclusive. Six studies compared drugs with nondrug interventions and showed consistently that stimulants, particularly MPH, may be more effective than nonpharmacological interventions. Twenty studies compared combination therapies with a stimulant or a nondrug intervention alone; no additional beneficial effects for combination therapies were shown. Nine studies compared tricyclic antidepressants with placebo and showed that desipramine may be more effective than placebo; no consistent effect was shown for imipramine. Fourteen studies (13 in school children and 1 in adults) evaluated long-term therapy (> or = 12 weeks) and showed a trend to general improvement regardless of treatment, but the length of followup was inadequate. MPH may reduce behavioral disturbance in children with ADHD while it is taken. Academic performance does not appear to be improved with stimulants. Twelve studies evaluated treatment in adults with ADHD. For MPH vs. placebo, the results were contradictory. Antidepressants may be effective in adults, but no beneficial effect was seen with pemoline, nicotine, or phenylalanine compared with placebo. Thirty-two reports (29 studies) evaluated adverse effects of drug therapy; many of the side effects associated with stimulant use appear to be relatively mild and of short duration and to respond to dosing or timing adjustments. Data are inadequate on the long-term effects and severity of adverse effects of most interventions. CONCLUSIONS: This report describes rigorous systematic reviews on the treatment of ADHD, ready for incorporation into evidence-based clinical practice guidelines or performance measures. The report also provides a detailed description of the many limitations of the evidence available and provides recommendations to fill existing knowledge gaps. Studies on ADHD have low reporting quality, methodological flaws, and heterogeneity across outcome measures and tests. A detailed description is included of the many limitations of the available evidence plus recommendations to fill existing knowledge gaps. Fulfilling such knowledge gaps will not be easy and will require genuine collaboration among decisionmakers. PMID- 10790992 TI - [Treatment and prognosis of testicular seminoma]. AB - Although testicular germ cell tumor is a relatively uncommon disease, it is a relatively common type of malignant tumor among young men. Seminoma accounts for approximately 50% of all germ cell testicular tumors. Since the vast majority of patients with seminoma present with early-stage disease and the disease responds well to treatment, almost all of the patients are cured. Patients who have stage I disease without obvious metastatic lesions have two treatment options, surveillance or adjuvant retroperitoneal radiotherapy, following inguinal orchiectomy. Stage IIA disease with a relatively small retroperitoneal lymph node metastasis is generally treated by retroperitoneal radiation therapy, although systemic chemotherapy with carboplatin is an alternative treatment. For patients with bulky retroperitoneal lymph node or distant metastases (stage IIB, III), systemic chemotherapy including cisplatin and etoposide appears to be the standard approach. Recently, 85% to 90% of patients with stage III disease are cured. Thus, the current therapeutic goal is cure of the disease with the minimum of treatment sequelae. PMID- 10790993 TI - [Management of non-seminomatous germ cell tumors of the testes]. AB - Since the advent of cisplatin-based chemotherapy in the 1970s, testicular cancer has become a model for curable cancer. We review the current treatment options for non-seminomatous germ cell tumors (NSGCT) of the testes. Surveillance is considered to be the most widely accepted option for stage I NSGCT in Japan. Because of the high relapse rate of about 30% in this setting, the establishment of a risk-adapted treatment protocol is warranted in stage I NSGCT. In early stage II NSGCT, both the primary RPLND and the primary chemotherapy show equivalent cure rates. These options give different balances of toxicity; namely, ejaculation disorders and drug induced infertility. In the treatment for NSGCT with good prognosis, both three courses of BEP and four courses of etoposide and cisplatin are appropriate and standard chemotherapy regimens. PMID- 10790994 TI - [Treatment for ovarian germ-cell tumors and the prognosis]. AB - This article reviews the major advances in therapy for malignant ovarian germ cell tumors. Most of the important developments such as greatly improved survival and the preservation of reproductive potential in most young patients have occurred during the past two decades. We will also describe the major unresolved issues in this field. PMID- 10790995 TI - [Extratesticular germ cell tumor focusing on treatment and prognosis]. AB - About 3% to 5% of all germ cell tumors are thought to be primary extratesticular tumors, including a small group of benign tumors. Though the testes are clinically normal, diagnosing whether the tumor is primary or metastatic is problematic. Developments in ultrasonography have enabled the detection of tiny testicular lesions, and ultrasonography is applied to determine the biopsy sites. Nevertheless, burned-out tumor and carcinoma in situ are not always easy to find. Extratesticular germ cell tumors are usually treated using methods similar to those for advanced testicular tumors; however, the results are discouraging. Only limited cases are suitable for a monotherapy of surgery, so a combined treatment of cisplatin based chemotherapy and surgery is prevalent. Against seminomas, radiotherapy is effective. In addition to the advent of such pharmaceuticals as G CSF and serotonin receptor antagonists, recent development of peripheral blood stem cell transplantation is expected to produce improved prognoses among patients burdened with this refractory disease. A prognostic factor-based staging system developed by the International Germ Cell Cancer Collaborative Group is reasonable and useful. Non-seminomas are classified as having good or intermediate prognoses if retroperitoneal tumors having no non-pulmonary visceral metastases show good or intermediate markers. A poor prognosis is associated with extratesticular tumors with mediastinal primary or non-pulmonary visceral metastases or poor markers. Seminomas with normal AFP are classified as having good or intermediate prognoses if no non-pulmonary visceral metastases are seen or discarded. No seminomas are classified in the poor prognosis group. PMID- 10790996 TI - [Salvage chemotherapy in refractory testicular cancer]. AB - A treatment strategy for patients with refractory testicular cancer who failed in the initial therapy has not been established. Patients with metastatic testicular cancer are treated with BEP (BLM, etoposide, CDDP) therapy, which is recognized as the standard first line chemotherapy regimen. About 80% of patients attain complete remission (CR) with BEP therapy and following salvage surgery. The patients who fail to achieve CR or have recurrences during the period of follow up will be candidates for salvage chemotherapy. Salvage chemotherapies include VIP (ETP, ifosfamide, CDDP) therapy or high dose chemotherapy with peripheral blood stem cell auto-transplantation (PBSCT); however, the effectiveness of these therapies is limited. Those who fail in these salvage therapies are treated with irinotecan hydrochloride (CPT-11) in Japan. Clinical trials with paclitaxel and gemcitabine have recently been started in the United States and Europe. Further investigations are necessary to develop more useful regimens with these novel anticancer agents for refractory testicular cancer. PMID- 10790997 TI - [Current status of "low-dose cisplatin-5-FU therapy" for solid tumors (2nd report)--from a nationwide questionnaire on its adverse effects]. AB - A nationwide questionnaire survey was carried out on low-dose cisplatin-5-FU therapy for solid malignant tumors (mostly stomach and colon cancer) regarding its antitumor as well as adverse effects. The regimen was defined as 4 weeks administration of cisplatin 2-5 mg/body, 5 days a week, which was used in 47% of 82 institutions studied. Cases were classified into an Ab regimen (86 cases) with cisplatin 3-5 mg/body, and a Bb regimen (122 cases) with cisplatin 6-30 mg (mostly 10 mg)/body, both of which were given 5-FU 300-500 mg/body daily. The antitumor effect (CR + PR) was shown to be slightly higher in the Bb regimen than in the Ab regimen, and was 35% overall. Very few adverse effects appeared with either regimen for mostly major digestive, hepatic and renal functions, except for myelodepressions (leucopenia and thrombocytopenia), which were more than grade 3 in only a few percent with the Ab regimen, and a little more than 10% with the Bb regimen. The advantages and disadvantages of the A/Bb regimens appeared to be counter-balanced in terms of their antitumor and adverse effects. Most institutions using the Ab regimen favored repeating the regimen cycle as much as possible. Those using more than 6 cycles accounted for about 20% of hospitals. PMID- 10790998 TI - [Combined effect of S-1 and CDDP as a modulator for colon 26 liver metastasis]. AB - S-1 is a novel oral anticancer drug, composed of tegafur (FT), gimestat (CDHP) and otastat potassium (Oxo) in a molar ratio of 1:0.4:1, based on the biochemical modulation of 5-fluorouracil (5-FU). In this study the combined effect of S-1 and low-dose CDDP as a modulator for colon 26 liver metastasis in mice was evaluated. In an experiment with S-1 (5 mg/kg/day: po) and CDDP (0.25 mg/kg/day: i.p.) for 14 days, the combined effects for both liver metastasis and tumor of spleen were not superior to those with S-1 or CDDP alone group. Body weight loss was not greater in the S-1 + CDDP group than in the control group. In an experiment with S-1 (5 mg x 2/kg/day: po) and CDDP (0.25 mg/kg/day: i.p.) for 7 days, the inhibitory effects of S-1 + CDDP of liver metastasis and tumor of the spleen were remarkable compared with the S-1 alone group. However a greater loss of body weight was seen in the S-1 + CDDP group than in other groups. This study suggests that low-dose CDDP might be a modulator of S-1 for colon 26 liver metastasis. Further study is needed to determine the optimum dose and duration of treatment. PMID- 10790999 TI - Theoretical considerations and in vitro concentration response studies with two human colorectal carcinoma cell lines. The rational experimental base for clinical studies in regional chemotherapy. AB - The theoretical advantage of regional vs. systemic chemotherapy was calculated, based on pharmacokinetic considerations. The relevance of exposure time and high local concentrations of chemotherapeutic drugs for regional chemotherapy was elucidated in time dependent concentration response curves with two human cell lines. The theoretical pharmacological advantage of regional vs. systemic chemotherapy was defined by the formula Rd = (AUCi. a. 1 + AUCi. a. 2)/(AUCi. v.) and in hepatic artery infusion is for adriamycin (ADM) 5.8-.6, cis-platinum (CDDP) 8, epirubicine (EPI) 6.3, 5-fluorouracil (5-FU) 22-58, mitomycin C (MMC) 4.6, mitoxantrone (NOV) 6.3. All drugs but 5-FUDR exerted concentration response behaviour in the cell line-experiments. In the cell lines cytotoxicity depended on exposure time so that concentration chi time products at (IC50), (c chi t (IC50)), were calculated to determine an optimal in vitro exposure time. Based on these results and clinical considerations, optimal clinical exposure times could be defined for regional chemotherapy. The results may be of high relevance for e.g. hepatic artery infusion at the lower and chemoembolization or intraperitoneal instillation at the higher test concentration, respectively. PMID- 10791000 TI - [Clinical efficacy of low-dose weekly docetaxel combined with oral 5'-deoxy-5 fluorouridine (5'-DFUR) in advanced or metastatic breast cancer: a pilot trial]. AB - Docetaxel (TXT) has been shown to be an up-regulator of human pyrimidine nucleoside phosphorylase (PyNPase) in tumors. We have tried to use the combination of low-dose weekly TXT with 5'-DFUR (LD + D) in patients with advanced or metastatic breast cancer. In this study, we compared the clinical efficacy of LD + D with that of conventional full-dose TXT (FD) and that of low dose weekly TXT (LD). Twenty-one patients received 3 or 4 cycles of FD 60 mg/m2 every 3 or 4 weeks (group I), 14 patients received 8 cycles of LD 20-30 mg/m2 every week (group II) and 25 patients received 8 cycles of LD 20-30 mg/m2 weekly with oral 5'-DFUR 600-1,200 mg per day (group III). The overall response rates of groups I, II and III were 29%, 29% and 52% (p = 0.24), respectively. Grade 3-4 neutropenia was observed in 91% of group I, 6% of group II and 3% of group III. Nausea was present in 27% of group I, 28% of group II and 40% of group III. Higher incidence of gastrointestinal symptoms was found in LD + D, but the symptoms abated when the doses of 5'-DFUR were reduced. Low-dose weekly TXT with oral 5'-DFUR produced a higher response rate, but less hematologic toxicity than full-dose TXT, suggesting that this combination therapy is clinically useful and may be effective for patients with advanced or metastatic breast cancer. PMID- 10791001 TI - [A pilot trial of high dose CEP (cyclophosphamide, epirubicin, cis-platinum) therapy in patients with advanced and recurrent ovarian cancer]. AB - We evaluated the feasibility of high-dose CEP (cyclophosphamide 750 mg/m2, epirubicin 90 mg/m2, cis-platinum 70 mg/m2) therapy, with granulocyte colony stimulating factor support every 21 days, in 18 patients with advanced and recurrent ovarian cancer. Ten patients (56%) received 6 cycles of this regimen as planned. Toxicities more than grade 3/4 on' the WHO scale of neutropenia and thrombocytopenia were observed in all cases. Nausea, vomiting, mucositis, malaise, alopecia, hepatotoxicity, and fever were common adverse effects. The average relative dose intensity of cyclophosphamide, epirubicin, cis-platinum was 0.77, 0.77, 0.79 respectively, and each RDI decreased in the last two cycles. These data suggest that this regimen could be performed safely with careful consideration on hepatotoxicity and thrombocytopenia. PMID- 10791003 TI - [Effect of radiotherapy for inoperable remnant and recurrent gastric cancer- focusing of improvement in quality of life]. AB - We investigated six cases of inoperable remnant and recurrent gastric cancer treated with radiotherapy with regard to quality of life (QOL). The radiation dose was from 22 to 70 Gy, with a mean dose of 46 +/- 18 Gy. Four cases could be evaluated. Two were PR and two were NC. No patients could ingest food before radiotherapy, but two became able to eat whole gruel, one half gruel and three water. The mean survival time was 5.5 months and prognosis was not improved. Subjective symptoms such as food passage disturbance were decreased by radiotherapy and QOL was improved. Radiotherapy for patients with inoperable gastric cancer seemed to be effective in improving QOL. PMID- 10791002 TI - [Effects of oral 5-HT3 antagonists on chemotherapy-induced emesis in patients with gynecologic cancers]. AB - The efficacy of an intravenous 5-HT3 antagonist (granisetron) and four oral 5-HT3 antagonists (granisetron, ondansetron, tropisetron and ramosetron) on chemotherapy-induced emesis were investigated in 21 gynecologic cancer patients (63 courses). The severity of emesis after chemotherapy was classified in 4 grades (0: none, 1: slight loss of appetite, 2: severe loss of appetite, but tolerable, and 3: untolerable). The effect of 5-HT3 antagonists was judged by both the score for the severity of the emesis and the frequency of vomiting. The four oral 5-HT3 antagonists were almost the same in efficacy for 5 days after chemotherapy. Oral 5-HT3 antagonists were almost equipotent to intravenous granisetron for JT (paclitaxel + carboplatin) therapy or T (paclitaxel) therapy for 5 days after chemotherapy. However, they were ineffective for CAP (cisplatin + adriamycin + cyclophosphamide) therapy. From these results, oral 5-HT3 antagonists were proved to have a sufficient anti emetic effect after chemotherapy in cases of JT or T therapy. However, in cases of CAP therapy, intravenous 5-HT3 antagonists were thought to be preferable for the control of emesis due to chemotherapy. PMID- 10791004 TI - [A case of successful management of nonresectable advanced cholangiocellular carcinoma by intermittent hepatic arterial infusion at home]. AB - An 82-year-old female was referred to our hospital because a 16 x 8 cm tumor detected in her liver by abdominal ultrasonography (echo, hereafter) and CT. The patient was diagnosed as having highly advanced cholangiocellular carcinoma (CCC) by abdominal angiography. Since excision of the tumor was impossible due to the patient's age, a reservoir was indwelled for intra-arterial injection into the liver. Continuous injection of 1,000 mg 5-FU over 24 hours was performed every 2 weeks using a portable disposable pump 70 times. The tumor has been markedly reduced since the start of chemotherapy, with a reduction rate (PR) of 98% over the 3 years until the present. Since the frequency of administration was low, only twice a month, the patient had few side effects despite her old age, and injections could be performed in the outpatient department. Usually, the prognosis for CCC is poor. However, the patient has maintained a good QOL with the periodic intra-arterial injection of the carcinostatic into the liver, and this treatment has had a strong antitumor effect. This chemotherapy is thus considered useful for CCC which can not be resected. PMID- 10791005 TI - [A patient with advanced gastric cancer who obtained downstaging and underwent radical surgery by neoadjuvant chemotherapy]. AB - The patient was a 74-year-old man with extremely advanced gastric cancer. A CT scan of the abdomen showed enlargement of many huge abdominal para-aortic lymph nodes. Neoadjuvant chemotherapy (NAC) was planned in order to reduce or eliminate the tumor. Two cycles of FLP combination therapy (5-fluorouracil, leucovorin, cisplatin) were given. After NAC, a CT scan revealed marked shrinkage of the No. 16 lymph nodes, and a distal gastrectomy with extended radical lymph node dissection including the No. 16 nodes was performed. The histological effect was judged to be grade 2. There were no viable cancer cells in the No. 16 lymph nodes. The FLP combination therapy as NAC was so effective that it induced downstaging from stage IVb to IIIb. PMID- 10791006 TI - [A case of multiple liver metastases from colon cancer successfully operated after intraarterial chemotherapy performed in home]. AB - A 60-year-old male patient had unresectable multiple liver metastases from a sigmoid colon cancer that had been resected, and thus hepatic arterial infusion therapy was planned. A heparin coated catheter was inserted from the left thoracoacromial artery to the proper hepatic artery. 5-fluorouracil (1,000 mg) was administered via the catheter 24 hours/week using an implantable vascular device and a small disposable pump in his home. After 59 weeks, the metastatic liver tumors had decreased remarkably in size, and all tumors could be resected completely through surgery. Three weeks after the operation, the same intraarterial chemotherapy was restarted to prevent the recurrence in the liver. However, the patient died of lymph node recurrences. The intraarterial chemotherapy is thought to be useful for neoadjuvant therapy in patients with inoperable liver metastasis from colon cancer. PMID- 10791007 TI - [Absence of cross resistance between an irinotecan plus cisplatin combination and raltitrexed (ZD1694) in two patients with advanced colorectal cancer]. AB - Two patients with advanced colorectal cancer were consecutively administered two different chemotherapeutic regimens. The first patient showed an initial response to combination treatment with irinotecan plus cisplatin, but then progressed. He subsequently responded to treatment with a novel thymidylate synthase (TS) inhibitor, raltitrexed. The second patient, who progressed after exhibiting a response to raltitrexed, subsequently responded to irinotecan/cisplatin combination therapy. In conclusion, no clinical cross resistance between the regimens of irinotecan/cisplatin combination therapy and raltitrexed was observed in these patients with advanced colorectal cancer. PMID- 10791008 TI - [A case of epithelial cancer of the alveoli which responded favorably to the additional administration of UFT for refractory cancer after administration of carboplatin and docetaxel]. AB - Epithelial cancer of the alveoli is considered to be a pulmonary non-small cell carcinoma which responds poorly to carcinostatics. In one case of epithelial cancer of the alveoli which metastasized to both lungs and caused breathing to deteriorate rapidly, chemotherapy was applied with 500 mg of carboplatin (CBDCA) and 90 mg of docetaxel (TXT). Although the tumor was reduced initially, it was found to have been aggravated again three weeks after the start of the chemotherapy. In the second and third courses of the chemotherapy, CBDCA and TXT were administered in the same dosage as in the initial course, but with the oral administration of UFT (600 mg/day). The results were favorable, as evidenced by the absence of recurring aggravation. Currently, the patient has been followed on an outpatient basis for over six months with the administration of UFT. Good QOL is being maintained without any repeated aggravation of the tumor. PMID- 10791009 TI - [A case of recurrent breast cancer with carcinoma erysipeloides responding to sequential therapy with docetaxel (TXT) and doxifluridine (5'-DFUR) accompanied by leucovorin (LV)]. AB - A left radical mastectomy was performed on a 53-year-old woman, diagnosed with left inflammatory breast cancer, after local arterial chemotherapy with cyclophosphamide (CPA), doxorubicin and 5-fluorouracil (5-FU). Adjuvant therapy was added with irradiation and ECF. Four months after the operation, a red eruption was detected on the left upper chest wall. The lesion was diagnosed by skin biopsy as a recurrent breast cancer with carcinoma erysipeloides. Tumor marker levels suggested the recurrent cancer was ECF resistant, so we changed the chemotherapy regime to a single dose of TXT. Although tumor marker levels and the skin eruptions improved at the beginning of the therapy, pleuritis carcinomatosa was found. We changed the regime again to a continuous dose of 5'-DFUR and LV for day 1 to 7. With this regime the clinical symptoms improved, and 2 courses of this modified FL therapy were carried out. After the therapy, the tumor seemed resistant to this modified FL therapy. Therefore, we tried a sequential therapy with TXT and the modified FL, which induced an improvement in clinical symptoms. Two years later, the patient died from the breast cancer. Therefore, we conclude that the sequential therapy may be beneficial in managing untreatable carcinoma erysipeloides of recurrent breast cancer. PMID- 10791011 TI - [Present status of developmental therapeutics for anticancer agents in Japan]. PMID- 10791010 TI - [Pain relief with alendronate therapy in a breast cancer patient with bone metastasis]. AB - A 66-year-old woman developed a bone metastasis from breast cancer to the sternum in September, 1997. She received alendronate therapy, consisting of biweekly intravenous administrations of 10 mg-alendronate 6 times and monthly 20 mg alendronate infusions 15 times. The first alendronate administration markedly alleviated her bone pain. She obtained complete pain relief after the 4th alendronate infusion. However, an elevation of tumor marker levels in serum without any pain increase forced us to treat her with medroxyprogesterone acetate and doxifluridine in addition to the alendronate therapy. With these therapies, she has shown an objective response (PR) of the bone metastasis for 8 months. In conclusion, alendronate therapy was effective against bone pain due to metastasis of breast cancer. PMID- 10791012 TI - Prevention and treatment of non-insulin-dependent diabetes mellitus. AB - The benefits of exercise training in the prevention and treatment of insulin resistance, impaired glucose homeostasis, and NIDDM are strongly supported by current research. The actual mechanisms involved have not been completely identified but occur at the systemic, tissue, and cellular levels. The adaptations that are responsible for the prophylactic effects of exercise training, however, start to subside rapidly once training ceases and are completely lost within 1 to 2 weeks of detraining [4, 17, 37, 68, 161]. Thus, the benefits of exercise training must be renewed on a regular basis. In addition, many of the systemic and cellular adaptations that are responsible for an improved skeletal muscle insulin action occur in only those muscles involved in the training program [4, 28]. Therefore, exercise training programs that consist of various modes of exercise, and which require the use of a large muscle mass, such as swimming, power walking, and strength training, may be the most advantageous for the prevention and treatment of insulin resistance and associated diseases. PMID- 10791013 TI - Cardiovascular consequences of exercise hyperpnea. AB - In summary, evidence shows that the respiratory muscles demand a significant portion of the cardiac output during maximal exercise. Estimates of both animal and human blood flow and VO2 to the respiratory muscles during maximal exercise approximate 14-16% of the total cardiac output and VO2. During heavy exercise, this metabolic demand of the respiratory muscles affects the distribution of cardiac output between the respiratory muscles and the legs such that leg vascular conductance and blood flow increases with respiratory muscle unloading and decreases with respiratory loading. The reflex effects underlying this blood flow redistribution remain unknown; however, these data do clearly support the existence of a significant sympathetic effect output to working skeletal muscle in heavy exercise. These data also invite the exciting (although speculative) prospect of important chemo- or mechano-induced reflexes emanating from respiratory muscle under load.Finally, while not yet completely understood or investigated, it appears that respiratory muscle work during strenuous exercise affects exercise performance. PMID- 10791015 TI - Moment arm of a muscle force. PMID- 10791014 TI - Skeletal muscle myosin II structure and function. AB - Recent experimental advances in structural biology, biophysics, and molecular biology have dramatically increased our understanding of the molecular mechanism of muscle contraction, as well as the assembly of myosin filaments. Future studies are required to detail, for example, the molecular cause of the conformational change during the power stroke and ATP hydrolysis, as well as the nature of the communication between nucleotide and actin binding sites. Based on the structural and functional homology between myosin and other molecular motors, these findings have implications not only for understanding muscle contraction, but for understanding numerous aspects of motility in all cellular systems as well. PMID- 10791016 TI - Exercise and pain: the neurobiology, measurement, and laboratory study of pain in relation to exercise in humans. PMID- 10791017 TI - Influence of body water and blood volume on thermoregulation and exercise performance in the heat. PMID- 10791018 TI - Integrative biology of nitric oxide and exercise. PMID- 10791019 TI - Rapid spinal mechanisms of motor coordination. PMID- 10791020 TI - Exercise training and cellular adaptations of normal and diseased hearts. PMID- 10791021 TI - Cardiovascular adaptations to endurance exercise training in older men and women. PMID- 10791022 TI - Transcriptional regulation in response to exercise. AB - Much progress has been made in recent years into understanding molecular mechanisms by which transcription is regulated following changes in physiological stimuli. This review has tried to focus on what is known about four specific physiological challenges--mechanical load, intracellular calcium, hypoxia, and redox state. Because of our biased interest in exercise, it was our goal to review these relatively well-studied systems so that we might provide insight into potential mechanisms that govern exercise-induced transcriptional changes. What becomes obvious, when reaching the end of this review, is that there are many common themes among the different physiological responses described. Some examples include the activation of IEGs, such as c-jun and c-fos, the phosphorylation of the transcription factor CREB, and the importance of the serum response element and the serum response factor. These commonalities across the different physiological systems suggest a certain redundancy or shared mechanism(s) for regulating transcription in response to physiological stimuli. While very little is known at this time about how exercise regulates transcription, it is an exciting time in this field of research. The recent growth in the molecular biological research literature of more physiologically based studies provides exciting new molecular and cellular tools for those researchers willing to take on the challenge of understanding the complex mechanisms of exercise-induced adaptations. PMID- 10791023 TI - RNA secondary structure and compensatory evolution. AB - The classic concept of epistatic fitness interactions between genes has been extended to study interactions within gene regions, especially between nucleotides that are important in maintaining pre-mRNA/mRNA secondary structures. It is shown that the majority of linkage disequilibria found within the Drosophila Adh gene are likely to be caused by epistatic selection operating on RNA secondary structures. A recently proposed method of RNA secondary structure prediction based on DNA sequence comparisons is reviewed and applied to several types of RNAs, including tRNA, rRNA, and mRNA. The patterns of covariation in these RNAs are analyzed based on Kimura's compensatory evolution model. The results suggest that this model describes the substitution process in the pairing regions (helices) of RNA secondary structures well when the helices are evolutionarily conserved and thermodynamically stable, but fails in some other cases. Epistatic selection maintaining pre-mRNA/mRNA secondary structures is compared to weak selective forces that determine features such as base composition and synonymous codon usage. The relationships among these forces and their relative strengths are addressed. Finally, our mutagenesis experiments using the Drosophila Adh locus are reviewed. These experiments analyze long-range compensatory interactions between the 5' and 3' ends of Adh mRNA, the different constraints on secondary structures in introns and exons, and the possible role of secondary structures in RNA splicing. PMID- 10791024 TI - Two novel regulatory genes, fliT and fliZ, in the flagellar regulon of Salmonella. AB - The flagellar operons of Salmonella are divided into three classes with respect to their transcriptional hierarchy. Expression of the class 2 operons requires the class 1 gene products, FlhD and FlhC, and is increased by mutation in the flgM gene, which encodes a class 3-specific anti-sigma factor. Here we report the identification of two novel regulatory genes for class 2 transcription. Presence of the fliZ and fliT genes on multicopy plasmids enhanced and inhibited, respectively, transcription from a chromosomal class 2 promoter. Disruption of the fliZ and fliT genes on the chromosome decreased and increased, respectively, class 2 expression. These results suggest that the fliZ and fliT genes may encode positive and negative regulatory factors, respectively, for class 2 expression. Enhancement of class 2 expression by the flgM mutation was cancelled by the coexisting fliZ mutation, indicating that FliZ is essential for this enhancement. PMID- 10791025 TI - Geographical variation of the alleles at the two prolamin loci, Pro1 and Pro2, in foxtail millet, Setaria italica (L.) P. Beauv. AB - Allelic variation at the two prolamin loci (Pro1 and Pro2) and its geographical distribution in 560 local cultivars of foxtail millet (Setaria italica) mainly from Eurasia were studied using SDS-polyacrylamide gel electrophoresis (SDS PAGE). Genetic analysis of a newly detected polymorphic band, band 6, indicated that it is controlled by an allele at the Pro2 locus, which was designated as Pro2f. Two alleles (Pro1a and Pro1null) at the Pro1 locus and six alleles (Pro2a, Pro2b, Pro2c, Pro2d, Pro2e and Pro2f) at the Pro2 locus were detected among the cultivars examined. Although the frequency of the Pro1a allele varied from 0% in the Nansei islands of Japan and Africa to 66% in Afghanistan, no apparent trend was observed in geographical distribution. In contrast, two common alleles at the Pro2 locus, Pro2b and Pro2f, had clear differential geographical distribution. The Pro2b allele was most frequent in Europe and decreased in frequency eastwards. The Pro2f allele occurred frequently in subtropical and tropical regions including the Nansei islands of Japan, the Philippines, Nepal, India, Pakistan and Africa. All eight alleles at the Pro1 and Pro2 loci occurred in China, suggesting China is a center of diversity. The origin of geographical differentiation of local cultivars into a "tropical group" characterized by the Pro2f allele and other genes was discussed. PMID- 10791026 TI - Identification and phylogenetic analysis of gypsy-type retrotransposons in the plant kingdom. AB - PCR was performed with degenerate primers which hybridized to the homologous sequences in the reverse transcriptase (rt) genes of gypsy-type retrotransposons from rice (RIRE3, RIRE8 and RIRE2), using total DNA samples from various plants (monocots, dicots, pine, ginkgo, horsetail, liverwort and algae) as templates. Cloning and sequencing showed that the amplified fragments had various degrees of homology to the rt sequences of rice retrotransposons. Phylogenetic analysis showed that these retrotransposon homologues and some additional gypsy-type retrotransposons previously identified from plants could be classified into two families, A and B. In each family, the retrotransposons were further classifiable into several subfamilies. Interestingly, retrotransposons from a single or related plant species were clustered in each subfamily. This indicates that sequence divergence during vertical transmission has been a major influence on the evolution of gypsy-type retrotransposons in plants. The retrotransposons isolated from one plant species could often be classified into the two families. This indicates that the gypsy-type retrotransposons of a family evolved independently within a species without affecting the evolution of retrotransposons of the other family. Retrotransposons in each subfamily are characterized by the lengths of LTR, by the nucleotide sequences in the terminal regions of LTRs, and by the PBS (primer binding site) sequence complementary to the 3' sequence of a particular tRNA species. PMID- 10791027 TI - N-terminal amino acid sequences of prolamins encoded by the alleles at the Pro1 and Pro2 loci in foxtail millet, Setaria italica (L.) P. Beauv. AB - N-terminal amino acid sequences of six prolamins encoded by seven alleles at two loci, Pro1 and Pro2, of foxtail millet (Setaria italica (L.) P. Beauv.) were analyzed and compared with other prolamins of subfamily Panicoideae. Based on the N-terminal amino acid sequences, band 3 (the prolamin purified from band 3) which is controlled by an allele at the Pro1 locus and bands 1, 2, 4, 5 and 6 which are controlled by alleles at the Pro2 locus could be classified into three groups. Band 3 was found to be homologous to the prolamin of pearl millet (Pennisetum americanum) and is designated as the "pennisetin-like prolamin". Bands 2 and 4, and bands 1, 5 and 6 were subdivided into "x-type prolamin" and "y-type prolamin". Both of the x-type and y-type prolamins showed homology with prolamin of Echinochloa crus-galli and alpha-zein-like prolamins of maize, sorghum and Job's tears. Therefore, these prolamins were designated as "alpha-zein-like prolamin". These results suggest that alleles at the Pro1 locus and those at the Pro2 locus have not arisen from an identical ancestral gene, and that the Pro2 locus comprise two tightly linked genes, which encode similar prolamins. Hypotheses on the diversification of alleles at the Pro2 locus are discussed based on the N-terminal amino acid sequences of the respective bands, combinations of bands controlled by the alleles, and frequencies of the alleles. PMID- 10791028 TI - [Detecting carriers of a deletion in the dystrophin gene in families with a single case of Duchenne/Becker muscular dystrophy]. AB - A search for female mutation carriers was performed in 40 families with an isolated case of Duchenne/Becker muscular dystrophy due to a deletion in the dystrophin gene. Intragenic restriction sites and microsatellite sequences (CA repeats) were analysed in females possible carriers of the deletion. Application of this approach enabled us the detection of the deletion in 19 females in 9 families and exclusion of the deletion in 41 females in 23 families. The results of DNA analysis in the remaining 8 families were not informative. PMID- 10791029 TI - [Carrier's detection in families affected by Duchenne/Becker muscular dystrophy in which DNA from affected individuals is not available]. AB - Carrier/noncarrier status of the mutated dystrophin gene was established in 9 females from four families with Duchenne/Becker muscular dystrophy, in which samples of DNA from the affected members were not available. Analysis of extra- and intragenic polymorphic segments of the dystrophin gene enabled identification of two female carriers and exclusion of carriership in four females. In three cases the results were not informative because of recombination in the analysed segment of the gene. PMID- 10791030 TI - [Platelet membrane fluidity and receptor exposition in patients with Alzheimer's disease]. AB - The majority of reports indicate that blood platelet membrane fluidity is increased in Alzheimer's disease (AD) patients. The increased membrane fluidity implies that platelet membrane receptors are less exposed to the external environment. To verify this hypothesis we tried to estimate platelet membrane fluidity at its two different depths and receptors' exposure after platelet activation in 12 AD sufferers. Platelet membrane fluidity was measured by the EPR with the use of 2 spin-labelled markers. In AD patients both "near surface" (p < 0.04) and "deeper depth" (p < 0.005) fluidity was significantly increased. The exposure of platelet granule membrane protein--P-selectin and membrane glycoprotein receptors: alpha subunit of glycoprotein Ib and beta3 subunit of GP complex IIb/IIIa were measured by flow cytometry with the use of human platelet monoclonal antibodies labelled with fluorescein and ficoerithrin. The exposure of GPIb alpha subunit was significantly decreased both in resting state (p < 0.0001) and after thrombin activation (p < 0.005). In thrombin-activated platelets the expression of P-selectin and beta3 subunit of fibrinogen receptor were also significantly decreased (p < 0.00001 and p < 0.04, respectively). The authors conclude that both platelet membrane fluidity and receptor exposure might serve as an adjunct marker of in vivo AD diagnosis. PMID- 10791031 TI - [The usefulness of MRI of the central nervous system in certain diseases of blood and hemopoietic system with neurological symptoms]. AB - The results are presented of MRI studies of the brain and spinal cord in patients with neurological symptoms in the course of certain diseases of the haemopoietic system (leukaemias, lymphomas). The analysis showed that MR images, although not specific, can be very useful for the evaluation of the intensity and topography of haematological lesions in the central nervous system. However, the diagnosis of these processes is not easy. In the differential diagnosis focal lesions of vasogenic origin, infections and radiation-induced lesions should be considered, and in cases of lymphoma--metastases are a possibility. PMID- 10791033 TI - [Is there any correlation between cerebral atrophy, EEG abnormalities and epileptic attacks in patients with multiple sclerosis?]. AB - Among 100 patients with MS, in 65 patients CT examination revealed cerebral atrophy (in 58 cortical and subcortical atrophy, in 7 patients only cortical). In 25 (25%) patients with cerebral atrophy EEG findings were focal type of paroxysmal activity. Only five patients with paroxysmal discharges had epileptic fits. Mean age in these 5 patients was 47 years, mean duration of the disease 17 years. This means that seizures in MS patients appeared in late period of the disease and no correlation was between cerebral atrophy and epileptic seizures. Paroxysmal discharges occurred more often in EEG examination, than clinical seizures. PMID- 10791032 TI - [Apomorphine in treatment of Parkinson's disease with fluctuations]. AB - Apomorphine is a non-specific dopamine agonist, most similar to it, with a strong action on D2, D3, D4 receptors and weaker action on D1 and D5 receptors. It has been known since 100 years, and in Parkinson's disease it was used first in 1970 by Schwab and Cotzias. Apomorphine is used in Parkinson's disease with high-grade fluctuations of symptoms which cannot be controlled by oral drugs, especially in off" periods resistant to levodopa. After subcutaneous administration it changes the "off" to "on" period within 5-10 minutes. Unfortunately, its effect is short lasting and wears off after 40-90 minutes. Apomorphine is administered in repeated single subcutaneous injections or in continuous subcutaneous infusion, if more than 7-9 single injections are required daily. Before beginning of treatment the optimal dose of apomorphine should be determined. For counteracting its emetic action domperidon (Motilium) is given additionally 20 mg t.d.s. Apomorphine produces no tolerance and is not losing its effectiveness with continued treatment. The most frequent adverse effects during long-term treatment are local cutaneous reactions, increased intensity of dyskinesia during the "on" period, visual hallucinations whose illusory character is clear to the patient, psychoses, orthostatic hypotension. The authors treated 8 patients with marked fluctuations in Parkinson's disease treated with levodopa. In 7 cases the effects was good--6 of them received 2-3 mg s.c. 3-4 times in 24 hours for 7-12 days. One patient has been treated 9 months with good result. In one case the intensity of dyskinesia made impossible treatment continuation. PMID- 10791034 TI - [Community-based rehabilitation of patients with nervous system damage]. AB - 495 patients with stroke were rehabilitated after discharge from hospital. From January 1st 1993 to December 31st 1997 they were included in community based rehabilitation (CBR) programme. We rehabilitated 12-14 patients monthly. In the therapeutic team were: physician, two physiotherapist and social worker. We received financial support from PHARE programme, National Fund of Rehabilitation for Disabled, Health Ministry and the local authorities. Now it is the most cost effective home-based rehabilitation system in Poland. PMID- 10791035 TI - [Brain temperature during craniotomy in general anesthesia]. AB - Mild hypothermia may occur spontaneously or, because of its putative neuroprotective effect, may be induced purposefully during neurosurgical procedures. Though the brain is the organ targeted for the purpose of neuroprotection, little is known about its temperature during general anaesthesia and craniotomy. The purpose of this study was to define the relations between core, skin and brain temperature during craniotomy and to compare two modes of inducing thermal insulation in patients during operative procedures. To achieve this we recorded core: rectal (Tre), oesophageal (Tes) and tympanic (Try) temperature, brain temperature in the subdural space (Tsd), and skin temperature on the thigh (Tfe), forehead (Tfr) and sternum (Tst) in 15 patients undergoing standard procedure for aneurysm clipping. In 13 patients the core temperature decreased, whereas skin temperature increased, after induction of general anaesthesia with isofluran. Nevertheless the mean body temperature remained unchanged, thus supporting the view that the cause of the resultant core hypothermia was heat redistribution between the thermal core and the periphery. Special thermofoil proved to be only as effective as a plain cotton blanket in preventing further heat loss during the later phases of the operation. Brain temperature was found to be the lowest core temperature throughout the procedure. It differed by as much as 0.1-1.2 degrees C from rectal temperature (mean 0.75 +/ 0.41 degree C) and reached the level of mild hypothermia (below 35 degrees C) even in those patients in whom rectal temperature indicated the state of normothermia. Furthermore tympanic and oesophageal temperature was on average 0.5 degree C higher than brain temperature. In conclusion, temperature measurements obtained in standard sites do not reflect brain temperature reliably during craniotomy and general anaesthesia. This indicates that the direct measurement of intracranial temperature is necessary for correct estimation of brain hypothermia. PMID- 10791036 TI - [8-oxoguanosine as a marker of neoplastic process in brain]. AB - Reactive oxygen species are toxic and cancerogenic factors to living organisms. They are suggested to cause DNA damage (modification) that triggers cancer development. It seems that oxidative damage product 8-oxo-deoxyguanosine (8-oxo dG) which induces transversion of G to T could be a good chemical marker for cancerogenesis. The aim of our studies was to use 8-oxo-dG as a probe for brain tumour in 17 patients operated on for intracranial neoplasm. Among the patients there were 7 female and 11 male aged from 14 to 60 year. Mean age was 42.88 +/- 16.14 yrs. Several types of tumours were selected histopathologically: from neuroepithelial tissue--6 cases, meningeomas--4, metastases--3, lymphomas--2, neurinoma--1 and chondrosarcoma--1. The tumour tissue was collected from removed material and stored at -20 degrees C. DNA from the neoplastic tissues was isolated by salt method. After hydrolysis of DNA with nuclease P1 and dephosphorylation with bacterial alkaline phosphatase, the mixture of nucleosides was analysed by liquid chromatography method connected with electrochemical detector (HPLC-ECD) working at potential 400 mV. We found higher level of 8-oxo dG in DNA of patients with malignant tumour (glioblastoma). However, at the present stage of these studies there was no proportional correlation between the level of 8-oxo-dG in DNA in tumour tissue and its malignancy. PMID- 10791037 TI - [Own experience with the treatment of intracranial aneurysms using "Perneczky" - Zeppelin clips]. AB - The basic design and the mechanism of clip application have not changed for many years. However in cases of aneurysms located deeply inside brain structure, in narrow spaces, multiple aneurysms or those which require applying of several clips the surgeon may have problems with visual control of the position of the clamping arms on the neck of aneurysm, nearby blood vessels and nerve structures. The above mentioned problems motivated prof. Axel Perneczky to construct a new model of clip with an applying and a removing holder whose mechanism is based on the principle of inverted--spring. Between February and August 1998 in the Neurosurgical Clinic in Katowice, 12 Perneczky's clips were applied in 9 patients. Intraoperative complications were not observed in any of the cases. In two cases of aneurysms located on the bifurcation of the basilar artery, application of two clips enabled precise intraoperative control of clip position. PMID- 10791038 TI - [Repeated operations for recurrent lumbar disc herniations]. AB - A study of 73 patients who underwent reoperation for persistent or new complaints following initial lumbar discectomy is presented. 32 patients showed true recurrence of disc herniation (6 on the contralateral side) and 41 at a different level. 14 patients needed 3rd operation. Final results were evaluated using the Functional Economic Rating Scale elaborated by Prolo. Mean score for group operated on twice was 8.1 according to Prolo and 7.1 for group operated on 3 times. PMID- 10791039 TI - [GABA-ergic system and antiepileptic drugs]. AB - gamma-Aminobutyric acid (GABA) belongs to the main inhibitory neurotransmitters in the central nervous system and activates three types of specific receptors- GABAA, GABAB i GABAC. At present, little is known about GABAC-mediated events. GABAB receptors are metabotropic, whilst stimulation of ionotropic GABAA receptors results in opening the chloride channel, followed by influx of chloride ions and hyperpolarization. The GABAA receptor possesses also binding sites for benzodiazepines and barbiturates which, via these sites, enhance GABAA-mediated events. Another antiepileptic drug potentiating GABA-ergic inhibition is valproate, which increases synthesis of GABA and reduces its metabolism. Among new antiepileptic drugs associated with the GABA-ergic system are tiagabine, vigabatrin, and to a certain degree--gabapentin. Tiagabine blocks neuronal and glial uptake of GABA whilst vigabatrin increases the synaptic concentration of GABA by inhibition of GABA aminotransferase. Gabapentin, probably through the activation of glutamic acid decarboxylase, leads to the increase in synaptic GABA. However, this antiepileptic drug also binds to specific sites within voltage-dependent calcium channels, which results in reduced intraneuronal concentration of calcium ions. Presumably, tiagabine and vigabatrin possess only one mechanism of action, associated with increased GABA-ergic inhibition. Although topiramate and felbamate were shown to enhance GABA-mediated events, they have additional mechanisms of action, including blockade of voltage dependent sodium channels and inhibition of glutamatergic neurotransmission. PMID- 10791041 TI - [Paroxysmal dyskinesis and epilepsy]. AB - Based on the available literature, we outline the classification criteria of paroxysmal involuntary movements with instant onset and termination. Four types of those movements are currently distinguished: dystonic Mount-Reback paroxysmal choreoathetosis, kinesigenic paroxysmal choreoathetosis, exercise-induced paroxysmal dystonia, and hypnogenic paroxysmal dystonia. We present the clinical characteristics of those entities, as well as diagnostic and therapeutic aspects. An association with epilepsy is emphasised, which seems to predominantly apply to hypnogenic dystonia and kinesigenic paroxysmal choreoathetosis. PMID- 10791040 TI - [Muscular dystrophies with dystrophin glycoprotein complex deficit]. AB - Progressive muscular dystrophies with primary or secondary dystrophin glycoprotein complex components' deficit are described. Some variants of limb- girdle and congenital muscular dystrophies as well as Duchenne/Becker muscular dystrophy belong to this group. The structure of dystrophin glycoprotein complex (DGC) which is localised within muscle sarcolemma is presented. Usefulness of the assessment of the particular DGC components is underlined and importance of the resulting prognostic as well as diagnostic implications are stressed. PMID- 10791042 TI - [Screw fixation of craniocervical junction]. AB - In recent years an increase has been observed of the use of screw techniques for the fixation of the craniocervical junction. For clinical use two techniques have been introduced: (1) transarticular screw fixation, and (2) transpedicular screw fixation. In the former the screw is inserted through the C2 lateral mass, the fissure of the C1-C2 joint, and the C1 lateral mass. (2) in the latter the screw is inserted into the C2 pedicle and anchored in C2 vertebral body. Transarticular or pedicle screws can be easily connected to longitudinal elements such as rods or plates, and combined with lateral mass screws of the remaining cervical vertebrae or occipital screws. In comparison to sublaminar wiring or interlaminar clamping the screw techniques: (a) strengthen the stiffness of the construct and speed up fusion, (b) allow fixation in the absence or deficiency of laminae as a result of trauma or laminectomy, and (c) can selectively include only the affected segments. Increased construct stiffness is due to deep anchorage of the screw in bone providing thus a solid grip on the vertebra. Both techniques require preoperative assessment of the course of the vertebral artery using imaging methods. In about 18% of cases abnormal course of the artery precludes screw use. Pedicle screw insertion requires direct control of the medial and superior walls of C2 pedicle with dissector introduced into the vertebral canal, which requires removal of the atlantoaxial ligament. Additional control can be achieved with lateral fluoroscopy. The entry point for transarticular screw is on the lateral mass of the odontoid 2-3 mm laterally to the medial margin of C2 facet and 2-3 mm above the C1/C2 articular fissure. The screw trajectory is 0-10 degrees in horizontal plane and towards the anterior C1 tuberculum in sagittal plane. PMID- 10791043 TI - [Primitive neuroectodermal tumor of rare localization in two members of one family]. AB - A 26-year-old man with primitive neuroectodermal tumour (PNET) is reported. The tumour originated from the cervical spinal cord and was resected partially. Few months later dissemination of the tumour to the meninges occurred. Familial history revealed that the first daughter of the patient had died in age of 14 months three years earlier of a tumour of the right cerebral hemisphere, also diagnosed as PNET. PMID- 10791044 TI - [Myelomatous meningitis: a case report]. AB - Multiple myeloma is characterized by the production of a monoclonal immunoglobulin. Myelomatous meningitis is a rare occurrence in multiple myeloma. The signs of meningeal myelomatosis include impairment of consciousness, cranial nerve palsies, convulsions and plasmatic cells in the cerebrospinal fluid. Atypical, plasmatic cells in the cerebrospinal fluid are an important finding for the diagnosis of meningeal myeloma. A combination of radiation therapy and chemotherapy is the usual treatment. Patients with meningeal myelomatosis can have a good response to treatment initially, but their prognosis is poor and death occurs within a few months. We present the case of a woman with myelomatous meningitis. Multiple myeloma was diagnosed by serum protein electrophoresis and bone marrow aspiration. This case and a review of the literature show that clinical manifestations of meningeal myeloma are non-specific. PMID- 10791046 TI - [Severe sensorimotor polyneuropathy in primary hyperparathyroidism]. AB - A case is reported of severe sensorimotor polineuropathy in primary hyperparathyreoidism caused by presence of parathyroid adenoma and cancer in a 63 years old man. The question is discussed of paraneoplastic syndrome. PMID- 10791045 TI - [A case of neurogenic dysphagia responding to nitrates]. AB - We described a 47-year-old man with ischemic stroke who developed a brainstem syndrome with persistent dysphagia. He was fed by the nasogastric tube placed intermittently by himself for almost 7 months after the stroke. Elective feeding via percutaneous endoscopic gastrostomy (PEG) was not accepted by the patient. All treatment attempts with benzodiazepines, antidepressants and spasmolytic agents were unsuccessful. Videofluoroscopic investigation revealed excessive and long-lasting spasm of the upper esophageal sphincter which was associated with the massive aspiration of the contrast. The patient dramatically improved after treatment with nitroglycerin and long-acting nitrates with almost complete recovery of normal swallowing. A strikingly good effect of nitrates in the treatment of oropharyngeal dysphagia is emphasized by the authors. PMID- 10791048 TI - [Report on the 17th Congress of the Polish Neurological Society "Advances in the treatment of nervous system disease", September 1-4, 1999, Lublin]. PMID- 10791047 TI - [A case of cervical myelopathy treated by "open-door laminoplasty]. AB - A case of cervical myelopathy is reported. In the treatment "open-door" laminoplasty was applied using titanium implants after laminoplasty for stabilization of the posterior bone block. The "open-door" laminoplasty is an easy and safe method making possible avoiding of such complications after laminectomy as instability, kyphotic deformity and development of scar compressing the spinal cord. PMID- 10791049 TI - [Report on the educational visit to the neurosurgical department at George-August University in Gottingen, Germany]. PMID- 10791050 TI - [Melanin: structure, biosynthesis, biological functions]. AB - Melanins are the group of natural black pigments. The structure of melanin macromolecules is irregular network arising from phenolic precursors in consequence of enzymatic and autooxidation. Melanin is stable polyradical, contain some semiquinone radicals and accumulate the exogene radicals and other active oxygen species, heavy metals, electrophyl toxic compounds. Some this properties determine the antioxidant, antitoxic, antiradiation and antitumour activity of melanins. On the base of natural melanins is possible creation of some effective prophylactic and curative preparations. PMID- 10791051 TI - Structural transitions in polyadenylic acid and hypothesis on biological role of its double-stranded forms. AB - Under various conditions poly(A) exists in different forms such as single stranded helix, two double-helical forms and others. The formation of double stranded helices is induced by adenine protonation. Under physiological ionic strengths they are formed at acidic pH, but under the same conditions methylated poly(A) has double-stranded structure at alkaline pH. Since the shift of adenine protonation pKa to alkaline region may be caused not only by chemical modification of poly(A) but also its interaction with proteins, it is quite probable that double oligo(A)-helices are formed in the living cell as well. In this article the hypothesis on possible biological role of poly(A) double stranded forms has been discussed in details. The models of involvement of double oligo(A)-sequences of RNA in such intracellular processes as termination of mRNA poly(A) tails synthesis and autoregulation of poly(A)-binding protein synthesis are suggested as an example. PMID- 10791052 TI - [Placenta--a source of biologically active substances]. AB - In the review there are some data displaying the scientific and patent literature presenting placenta as a rich source of some biologically active substances (BAS): proteins, lipids, enzymes, glycolipids and glycolipoproteins, hormones. This information is capable to be used while developing the techniques of BAS producing from placenta in order of creating the organospecific complex preparations with medically-preventive properties. PMID- 10791053 TI - [Correction of the activity of certain enzymes in the rat liver mitochondrial electron transport chain by derivatives of alpha-tocopheryl acetate in toxic damage to the liver]. AB - Correcting action of vitamin E and it's short chain derivative on the activity of some mitochondria electron transport chain enzymes were investigated on models of acute and chronic toxic hepatitis. Inhibition of NADH- and succinate-cytochrome c oxidoreductase complexes activity was established in short term action of xenobiotics. Treatment of rats with CCl4 during 60 days lowered activity of NADH cytochrome c oxidoreductase complex and significantly increased activity of succinate-cytochrome c oxidoreductase complex and succinate dehydrogenase. Obviously, as a result of long term influence of hepatotoxic agents switching over in rat mitochondria electron transport from NAD-dependent way of substrate oxidation to succinate-dependent way took place. This event could be a part of the body adaptation mechanisms. Vitamin E and its short chain analogue corrected activities of investigated enzymes of mitochondria liver in the animals with acute and chronic hepatitis. PMID- 10791055 TI - [Kinetic mechanisms of linoleic acid oxidation by 5-lipoxygenase from Solanum tuberosum L]. AB - Linoleic acid oxidation by 5-lipoxygenase from Solanum tuberosum has been studied as affected by sodium dodecylsulfate (Ds-Na). The reaction system consisted of 5 lipoxygenase and mixed micelles of linoleic acid and Lubrol PX. It contained varying amounts of the enzyme effector--Ds-Na. The enzyme showed a pronounced cooperativity, and the reaction was governed by the Hill equation with h = 3.7. On the other side, increasing amounts of Ds-Na added to the system caused a tremendous increase of enzyme activity and simultaneous decline of h, with was proportional to Ds-Na concentration. Ds-Na had dual effect on 5-lipoxygenase- there was an optimal concentration of the compound (0.34 mM Lubrol PX; 0.2 mM LA; 0.13 mM Ds-Na; pH = 6.3) causing the 4-fold highest activation and h = 1.6. The further increase of Ds-Na led to the enzyme inhibition. If Ds-Na was 0.5 mM, h became 1. At this point, each molecule of 5-lipoxygenase bound 3 molecules of Ds Na and 1 molecule of linoleic acid, thus the total number of occupied binding sites was 4. A kinetic scheme of 5-lipoxygenase reaction has been proposed. It was found that the enzyme's kinetic behaviour could be explaine if assumed an existence of a special noncatalytic binding centre capable of binding several (up to 3) molecules of either substrate, or effector. Such a centre can serve as an anchoring site facilitating the enzyme binding to the surface of lipid aggregates containing insolubilized substrate molecules. Replacing linoleic acid in the binding site, Ds-Na activates the enzyme, possibly due to the much more effective translocation of 5-lipoxygenase to the surface of lipid aggregates. This mechanism can be an universal alternative to the FLAP-type regulation of 5 lipoxygenase activities. PMID- 10791054 TI - [Antioxidant properties and membranotropic effect of certain derivatives of 1,4 dihydropyridine]. AB - Interaction of alpha-tocopherol and 1,4-dihydropyridine with endoplasmic reticulum membranes and model systems, human serum albumin and phospholipid bilayer, was studied using the microcalorimetry and fluorescent probes procedures. Dependence of microviscosity changes in the endoplasmic reticulum membranes on the place of antioxidants localization (protein structures or phospholipid phase) was shown. Increase of membrane structuralization under the influence of 1,4-dihydropyridines blocked their antioxidant action in spontaneous and induced lipid peroxidation. PMID- 10791056 TI - [Effect of aromatic compounds and Mn2+ on the ligninolytic enzyme complex from the fungus Pleurotus floridae (FRIES) Kummer--a white-rot wood fungus]. AB - The influence of aromatic compounds and Mn ions on activities of ligninolityc enzymes from white-rot fungus Pleurotus floridae has been studied. The specific inducers: vanillic acid and vanillyl alcohol--for activity of manganese-dependent peroxidase; vanillyl alcohol--for activity of cellobiose: quinone oxidoreductase during submerged, fermentation of Pleurotus floridae in Kirk's medium have been revealed. The inducers of laccase activity among studied aromatic compounds have not been revealed. The influence of Mn2+ in concentration range 0.4-68.4 mM on activities of ligninolytic enzymes of submerged culture of fungus P. floridae has been studied. Concentration of Mn ions 32.4 mM was optimal for manganese dependent peroxidase activity. PMID- 10791057 TI - [Interaction of thiamine with rat brain synaptosomes]. AB - Thiamine has been shown to be bound specifically by a synaptosomal plasmatic membrane and transported inside to the nervous ending. Apparent K[symbol: see text] and Km for processes of binding and transport have been determined as equal 2.34 +/- 0.55 MKM and 3.92 +/- 1.3 MKM, respectively. The thiamine uptake by the isolated nervous endings (synaptosomes) at its physiological concentration is reduced in presence of metabolic inhibitors and partially depends on Mg2+ and Ca2+ ions, that can testify about the interrelation between endogenic thiamine phosphorilation and its transport through the membrane. Thiamine binding with synaptosomes is inhibited by ouabain and neurotoxins such as, latrotoxin and most significantly--with veratridin; tetrodotoxin fail to be efficient practically. In the conditions of synaptic membranes depolarisation their ability to bind thiamine is reduced and output of already uptaken with synaptosomes thiamine is observed. PMID- 10791058 TI - [Role of monoaminergic system in the mechanisms of developing a dependence on morphine]. AB - The influence of morphine chronic administration on the neuromediators contents in various structures of brain has been studied. It has been shown that the development of physical dependence on the morphine and abstinence syndrome is accompanied by reduction of activity of serototinergic system, which is manifested in decrease of serotonin and its precursor tryptophane levels. Adrenergic and dophaminergic systems do not react to the development and abstinence by changes of the contents of monoamines that testifies of interaction of opioid and neuromediator systems disturbance. PMID- 10791059 TI - [Characteristics of H2O2-initiated oxidation of blood serum lipids by kinetic parameters of chemiluminescence]. AB - H2O2-initiated free radical oxidation of blood serum lipids was investigated by the chemiluminescence method. The first flash of the chemiluminescence was stimulated by H2O2 decomposition on the reaction, similar to Fenton reaction. The time of the chemiluminescence flash second maximum was correlated with the contents of antioxidants. This dependence had a linear kind and was characterized by the correlation coefficient--0.9898. With increases of concentration of such antioxidant as a hydroquinone, the time of chemiluminescence flash second maximum grew. PMID- 10791060 TI - [Lipid peroxidation and Ca2+,Mg2+-ATPase activity of uterine myocyte cell membranes in hyperplastic processes]. AB - It was shown that intensity of endogenous, Feascorbic- and NADPH-dependent lipids peroxidation in the border zone was 2-2.5-fold as higher as in the normal tissues. In the central part of the knot the intensity of all these above mentioned forms of lipids peroxidations is much lower than in the normal tissue. Plasma membrane Ca2+, Mg(2+)-ATPase activity of the border zone of myomic knot is 2-fold as lower as in the normal tissue. 10(-6) M malonal dialdegide decreases the Ca2+, Mg(2+)-ATPase activity up to the level of this enzyme activity in the border zone of myoma knot. PMID- 10791061 TI - [Characteristics of the effect of implantation of the human ApoA1 gene on the condition of hepatocytes in rats of varying age]. AB - The results obtained show the essential changes in functional state of hepatocyte's plasmatic membrane due to the implantation of human ApoA1 gene to the rat liver. The changes in phospholipid composition, hyperpolarization, increase in activity of membrane bound enzymes, cytochrome P-450 and biosynthesis of liver total proteins have been found. The essential changes characterizing cell effect were more marked in the adult rats, and membrane effect in the old ones. PMID- 10791062 TI - [Effect of the antineoplastic substance brotheophine on the insertion of precursors into nucleic acids and proteins from Plisse lymphosarcoma]. AB - The effect of antitumor medicine Brotheophine to insertion of the marked precursors into DNA (14[C]-timidine), RNA (3[H]-orotic acid) and proteins (14[C] leucine) of tumor cells (Pliss lymphosarcoma) as the indices of biosynthetic processes was investigated. It has been shown that Brotheophine acts as an antimethabolite of purine exchange and inhibits the vertical genetic information transfer in tumor cell (DNA-RNA-protein). Namely, a significant inhibition of 14[C]-timide insertion to DNA has been observed. less distinct is the inhibition of 3[H]-orotic acid to RNA und 14[C]-leucine to proteins. The above revealed allows to assume that during Brotheophine treatment certain changes in DNA biosynthesis take place leading to synthesis of slightly transformed RNA with subsequent impairment of proteins synthesis. PMID- 10791063 TI - [Effect of alcohol intoxication and aldehyde dehydrogenase inhibitors on lipid peroxidation in rat liver]. AB - A single intraperitoneal administration of ethanol (3.5 g/kg) to rats induced a marked increase in lipid peroxidation and a decrease of antioxidative activity in the liver after 1 h when assessed by chemi-luminescence in liver homogenates. The pretreatment with aldehyde dehydrogenase inhibitor, disulfiram (200 mg/kg 24 hr before ethanol), caused a 10-fold elevation of the blood acetaldehyde levels, with no effect on the hepatic lipid peroxidation compared to control. Cyanamide (50 mg/kg, 2 h before the ethanol) increased approximately 100-fold the acetaldehyde levels, however, the changes in lipid peroxidation were not significantly different from that produced by ethanol alone. The present results suggest, that the metabolism of acetaldehyde and not acetaldehyde itself is responsible for the in vivo activation of lipid peroxidation during acute alcohol intoxication. Disulfiram prevents the ethanol-induced lipid peroxidation in the rat liver. PMID- 10791064 TI - [Investigation of oxosubstances in plant leaf lipids]. AB - Surface lipids of plant leaves of different groups were investigated by spectral, chromatographic and functional analysis methods. Absence of oxosubstances in surface lipids of coniferous and their characteristics in tropical species were shown. Functions of oxosubstances are discussed. PMID- 10791065 TI - [Some biochemical indices in chicken skin tissue during ontogenesis and under the effect of ionizing radiation]. AB - The research shows some biochemical indices in ontogenesis in broiler hen skin tissues and also its reaction to inner introduction of different doses of radioactive 137Cs. Namely, the irradiation ionizing irradiation influences actively to the indices of nuclein changes with 1 day and 1 week aged chickens. The quantity of phosphorus of nuclein acids is less with the chickens of the third group (500 Bk pr day) than with the second group (3000 Bk/day). The quantity of soluble protein, activity of the estimated fragments, the quantity of inorganic phosphorus, the concentration of ions of magnesium with chickens of the controlled group is larger than in the studied ones. It proves the influence of radionucleid 137Cs on indices which characterize the protein-nuclein change. PMID- 10791066 TI - [Change in the structure of rat liver mitochondrial membranes under the effect of ionizing radiation]. AB - The effect of gamma-irradiation (under 0-10(3) Gy) on the intensity of lipid peroxidation, the microviscosity of annular and free lipids and the polarization of membrane proteins tryptophan fluorescence was studied in the outer and inner mitochondrial membranes. Some specific individual peculiarities of the mitochondrial membranes post-radiation changes were established. PMID- 10791067 TI - [Lipid peroxidation in chicken digestive organs during postnatal ontogenesis under the effect of radioactive cesium]. AB - The activity of superoxide dismutase, catalase and content of malondialdehyde, hidroperoxide lipids was studied in the tissues of liver, duodenum, pancreas of 1 day and 2-10 weeks age chicken. It was found, that tissues of 1-day chicken ages characterized by high contents of products of hydroperoxide lipids and activity of superoxide dismutase. The content of lipid peroxide oxidation in chicken tissues has an age peculiarity. The insertion of radioactive ceziy acts as an antioxidant and influences the lipid peroxide oxidation. PMID- 10791068 TI - [Radiation-induced changes in the genome of pine seeds, forming under conditions of chronic irradiation in the Chernobyl exclusion zone]. AB - Comparative study of long-term existing genome damages in the chronically irradiated pine seeds from different parts of Chernobyl zone have been performed by a pulsed field gel electrophoresis and hydroxyapatite chromatography. Some differences were established for DNA degradation kinetic from protoplasts with various accumulated doses of irradiation. Results obtained could testify the existence of long-living changes in the experimental plants genome. PMID- 10791069 TI - Preparation of fibrin des-AA by thrombin. AB - The active thrombin is formed in the blood stream when the blood coagulation system is activated. It attacks fibrinogen, splits off two fibrinopeptides A and fibrinogen is transformed into des-AA fibrin which is able to polymerize spontaneously forming protofibrils. At high thrombin concentration the enzyme splits off two fibrinopeptides B and des-AA fibrin units are transformed into des AABB fibrin. These two forms of fibrin are widely used in the biological experiments. However des-AA fibrin is obtained usually from fibrinogen using the snake poisons (such as reptilase). Des-AA fibrin was obtained also by physiological enzyme thrombin, but that des-AA fibrin samples had the contamination of des-AABB fibrin. At the present paper we have described the method of the des-AA fibrin preparation by thrombin without any contamination of des-AABB fibrin. PMID- 10791070 TI - [Val709-Glu724 streptokinase binding site on plasminogen interacts with streptokinase sequence Thr361-Arg372 during plasminogen-streptokinase complex formation]. AB - Localization of the human plasminogen binding site on the streptokinase of complementary Val709-Glu724 plasminogen being crucial one in providing for the plasminogen streptokinase complex activity has been investigated. Experiments were performed with streptokinase fragments and synthetic decapeptides, antiplasminogen monoclonal anti-body IV-1c and synthetic peptide corresponding to Val709-Gly718 sequence of human plasminogen. It was found that plasminogen sequence Val709-Glu724 interacted with Thr361-Arg372 sequence of strepto-kinase. PMID- 10791071 TI - [Catalytic properties of Glu-plasminogen in a complex with monoclonal antibody IV 1c]. AB - Earlier it was shown that anti-plasminogen monoclonal antibody IV-1c was able to induce a catalytic activity in plasminogen. IV-1c activates plasminogen by binding to plasminogen protease domain with antigen binding site and to lysine binding sites by C-terminal lysines of gamma-chains. The effect of plasminogen and IV-1c concentration on rate of catalytic activity induce in Pg-IV-1c complex has been investigated. It was found that IV-1c inhibited an activation reaction at concentrations higher of equimolar to Glu-Pg. Glu-Pg did not inhibit reaction of activation in higher to IV-1c concentrations. Role of IV-1c gamma-chain C terminal lysine concentration in Pg activation is discussed. PMID- 10791072 TI - [Effector action of ethanol on the accumulation of Ca2+ in intracellular structures of uterine smooth muscle]. AB - The work is devoted to the investigation of ethanol direct effect on the transmembrane Ca2+ metabolism in the intracellular structures of myometrium. In the experiments in vitro it has been shown that the Mg2+, ATP-dependent system for Ca2+ accumulation in endoplasmic reticulum is more sensitive then Ca(2+) accumulating system in mitochondria. It has also been found that the oxytocin insensitive part of Mg2+, ATP-dependent Ca2+ accumulation of the endoplasmic reticulum is less resistant to ethanol inhibition than the oxytocin sensitive one. The data above revealed allow to discuss mechanism of ethanol action on the intracellular Ca2+ homeostasis in myometrium. PMID- 10791073 TI - [A new type of electrochemical immunosensor based on ion-selective field effect transistors]. AB - A new type of an electrochemical sensor based on the use of ion-selective field effect transistors (ISFETs) and conjugates of horse radish peroxidase with specific monoclonal antibodies was developed for the express determination of myoglobine in a solution. For this purpose a simple method of covalent immobilization of myoglobine on the surface of ISFET gate was worked out, an appropriate biochemical approach which allowed potentiometrical registration of the peroxidase activity was used, and an immune chemical analysis was accomplished in competitive way. It was shown that the sensitivity of the analysis with the help of the electrochemical immune sensor corresponds to the demands of medical practice to reveal early stages of myocardial infarction. This sensitivity was significantly higher then that which can be obtained by the traditional ELISA-method. Moreover, overall time of the analysis by the immune sensor was almost one order shorter than this by the ELISA-method. It is concluded that the proposed electrochemical immune sensor based on the ISFETs was very perspective for the express analysis of the level of different types of antigens and antibodies. PMID- 10791074 TI - [Distribution of N-([1-14C]-palmitoyl)ethanolamine in rat tissues]. AB - N-([1-14C]-palmitoyl)-ethanolamine distribution was studied in the rat tissues. The following sequence of the label inclusion into tissues by the way of decreasing the radio activity: adrenal >> diaphragm > spleen > kidney > testis > lung > liver > heart > brain > plasma > erythrocytes was obtained. PMID- 10791075 TI - [Current and future aspects of mitochondrial diseases]. AB - Though mitochondria have been a major source of energy production in eukaryotae since 15-20 billion years previously, existence of disorders due to primary abnormalities of their DNA has not been known until very recent years. In 1962, Luft et al reported the first case of such myopathy, and another case reported in 1967 by Shy et al was also the first case of generalized disorder with mitochondrial abnormalities. Since then, many case reports have followed including MELAS and other encephalomyopathies. Finally, in 1989, deletion of mitochondria DNA was found by Folt et al. Today, these disorders were able to be classified as follows: 1) LHON and A1555G type deafness as strictly limited non syndromic type, 2) encephalomyopathies and their incomplete forms due to common and other deletions of mitochondria DNA, 3) encephalomyopathies and their incomplete forms including MIDD, diabetes mellituis, cardiomyopathy, deafness due to point mutations of mitochondria DNA related MELAS and others, 4) Neurodegenerative types including Parkinson's disease, Alzheimer's disease, cerebellar degeneration, and amyotrophic lateral sclerosis, or neurologic disorders mimic to such diseases, 5) Mitochondrial involvement not due to primary abnormalities of mitochondria DNA. Possible mechanisms were discussed, but sufficient knowledge is lacking so far to clarify pathophysiology of these disorders and the role of deleterious DNA in aging. Possible effective therapeutic strategies were also discussed, but further development of research works on these disorders in the 21st century are needed to answer these questions. PMID- 10791076 TI - [Pathogenesis and treatment of neuropathy--recent advances]. AB - Wide variety of underlying mechanisms are present for pathogenesis and pathophysiology of neuropaties, some of the which are being elucidated recently. Remarkable recent advances are discovery of causative gene mutations of various hereditary neuropathies and elucidation of pathogenic mechanism of inflammatory demyelinating neuropathy. Particularly, causative gene mutations for Charcot Marie-Tooth (CMT) disease and pathogenic role of glycolipid antigens for Guillain Barre syndrome have recently been documented. Moreover, efficacious treatment based on the background pathophysiology have also been developed, such as intravenous immunoglobulin infusion and plasmapheresis for inflammatory demyelinating neuropathy, and liver transplantation for familial amyloid polyneuropathy. These advances on the understanding of pathogenic mechanisms and development of treatment have changed the clinical approach to peripheral neuropathies. In this review, I overviewed the recent advances on three neuropathies, familial amyloid polyneuropathy, chronic inflammatory demyelinating polyneuropathy, and Charcot-Marie-Tooth disease with special reference to pathogenic mechanism and treatment. PMID- 10791077 TI - [Vascular dementia]. AB - The vast majority of senile dementia consists of Alzheimer's disease and vascular dementia. However, there appear to be geographic differences in the world in the ratio of these two disorders and the types of vascular dementia prevalent in the regions: multi-infarct dementia, lacunar infarct dementia or Binswanger disease. Partly because of those factors, there have been considerable confusions in the classification and diagnostic criteria of vascular dementia. In order to clarify the confusions, it is necessary to recognize the presence of both dementias caused by multiple infarcts as the results of atherothrombosis and embolism, and those caused by multiple lacunar infarcts and periventricular white matter ischemic lesions. Since the clinical manifestations of different types of vascular dementia may be different, it is also necessary to establish the diagnostic criteria which is applicable to all different types of vascular dementia. In order to comprehend the clinicopathologic characteristics of vascular dementia, it is always necessary to analyze the location of arterial occlusion and the mechanism leading to the arterial occlusion. PMID- 10791079 TI - [Neuropathology of amyotrophic lateral sclerosis--from basic findings to topics]. AB - Amyotrophic lateral sclerosis (ALS) is a relentlessly progressive neurodegenerative disease that selectively affects upper and low motor neurons (UMNs and LMNs). The remaining LMNs show, in addition to normal appearance, a variety of cell pathology such as central chromatolysis, atrophy of the cell body and processes, Bunina bodies, and ubiquitinated hyaline and skein-like inclusions. Ultrastructural studies of the last two profiles indicate their processing by lysosomes. Whether apoptosis is responsible for motoneuronal death in ALS or not remains to be determined. In Kluver-Barrera (KB) staining, the pyramidal tracts in ALS usually appear normal at the pons in spite of their obvious pallor at the lower levels, leading to the dying-back hypothesis. However, axonal staining in such cases reveals obvious loss of large axons in the pontine pyramidal tracts, a finding inconsistent with the hypothesis. Similarly, in cases of motoneuron disease with the pyramidal tracts well stained by KB method, therefore, 'spinal progressive muscular atrophy (SPMA)', axonal staining demonstrated patent loss of large axons there. Thus, reappraisal seems to be required for previously reported SPMA cases, whose pathological diagnosis used to be made by KB staining alone. PMID- 10791078 TI - [The recent advances of HAM/TSP research]. AB - The ninth international conference on HTLVs and related disorders was held on April 5-9, 1999 at Kagoshima, Japan under the conference chairperson, Dr. Mitsuhiro Osame. In this meeting, world-wide epidemiological data on HTLV-I carriers, ATL patients, and HAM/TSP patients were summarized as shown in the table. The total number of them was supposed to be more than 2.2 millions, 1,200, and 3,000, respectively. To elucidate the localization of HTLV-I proviral DNA directly, double staining using immunohistochemistry and PCR in situ hybridization in the spinal cords of HAM/TSP patients were performed. HTLV-I proviral DNA was localized only to OPD 4-positive cells (Matsuoka et al, 1998). The localization of HTLV-I messenger RNA was the same (Moritoyo et al, 1996). A novel technique to detect HTLV-I tax protein was also developed. In HAM/TSP patients, 0.04-1.16% of the CSF cells and 0.02-0.54% of PBMCs were positive for HTLV-I tax protein (Moritoyo et al, 1999). It was also hypothesized that HLA alleles control HTLV-I proviral load and thus influence susceptibility to HAM/TSP. Two hundred and thirty-two cases of HAM/TSP were compared with 201 randomly selected HTLV-I seropositive asymptomatic blood donors. It was shown that, after infection with HTLV-I, the class I allele HLA-A*02 halves the odds of HAM/TSP (p < 0.0001), preventing 28% of potential cases of HAM/TSP. Furthermore, HLA-A*02 positive healthy HTLV-I carriers have a proviral load one-third that (p = 0.0114) of HLA-A*02 negative HTLV-I carriers. An association of HLA-DRB1*0101 with disease susceptibility was also identified, which doubled the odds of HAM/TSP in the absence of the protective effect of HLA-A*02 (Jeffery and Usuku et al, 1999). PMID- 10791080 TI - [The significance of the study of aphasia, apraxia and agnosia at present time]. AB - Neuropsychology initiated by the study of aphasia, apraxia and agnosia had been regarded as one division of psychiatry. This is one of the reasons why neuropsychology is minor territory in neurology. Some people say that neuropsychology seems difficult to understand, not scientific, not useful and is rather a man's taste than a medicine. I try to respond in this paper to these critics by way of analyzing our own cases while insisting on the today's importance of the study of aphasia, apraxia and agnosia. Our personal cases consist of the following three categories. 1) Primary progressive apraxia: proposal of the symptomatic concept suggested by the investigation of our own eight cases and those in the literature. 2) Musical alexia with agraphia in a trombone player: study of the cerebral localization in a person with special ability. 3) Disturbance of reaching proposal of intracerebral pathways in kinesthesia. Therefore it is highly possible that the significance of the study of aphasia, apraxia and agnosia should become enlarged now and after; and it seems urgent to answer its need that younger neurologists participate in this area. PMID- 10791081 TI - [Myotonia and channelopathy]. AB - Myotonia occurs with abnormalities of the muscle membrane, and there have been reports of various channel abnormalities including Cl, Na, and Ca channels. Historically in 1966, Lipicky and Bryant found the abnormality of the muscle Cl channel in Thomsen's disease and hereditary myotonia of goat. Thereafter, Na channel abnormality was found in myotonia associated with familial hyperkalemic periodic paralysis and paramyotonia congenita by using patch clamp technique. Myotonic dystrophy is the most common myotonic disorder and genetic studies disclosed the presence of abnormal triplet repeat in the chromosome 19q13.1 and the gene product is known as myotonin protein kinase. However, pathophysiological mechanism of myotonia in myotonic dystrophy is not yet clear except that there are mild decrease of Cl conductance in 5 out of 8 muscle fibers and late opening of Na channels also occurs. As for the treatment of myotonia, medications which work as Na channel blockers have been used and they are effective to reduce myotonia, but they do not improve muscle weakness. Recently dehydroepiandrosterone sulfate which is a male hormone secreted from the adrenal cortex has been used for myotonic dystrophy and the drug reduces myotonia and improves activities of daily living. The author confirmed that myotonia can be induced in the muscle preparations of mice by either anthracene-9-carboxylic acid (Cl channel blocker), or by anemone toxin (blocker of inactivation of Na channel), and dehydroepiandrosterone sulfate can reduce myotonic bursts in both of these muscle preparations by intracellular recordings. PMID- 10791082 TI - [Treatment of myasthenia gravis]. AB - The strategy of myasthenia gravis has been progressed during recent 30 years. Extended thymectomy and alternate-day prednisolone administration remarkably improved therapeutic effect in myasthenia gravis. Besides of these therapies, plasmapheresis and immunoadsorption plasmapheresis extremely improved mortality in myasthenia gravis. 1. Corticosteroid therapy Usually initial doses of PSL is 20-30 mg every other day in the morning after thymectomy, and increased 5 mg every on-day. Maximum PSL doses is 80-100 mg alternate-day, then this doses will be maintained over 4 weeks. After these procedures, PSL is gyradually decreased and discontinued within 2-3 years. 2. Thymectomy Extended transsternal thymectomy is the most useful method, because many authors have reported that re-thymectomy reveals re-appearance of thymus. 3. Invasive thymoma Although this type of thymoma has been called malignant thymoma, it should be called as invasive thymoma since any thymoma can invade surrounding tissues for a long subclinical period. Strategy of treating invasive thymoma is PSL administration, radiation and chemotherapy. Chemotherapy should be performed depend on histopathological findings of thymoma. CHOP therapy is useful for the lymphocytic cell predominant type of thymoma, however CHOP + cis-platin therapy is useful for the epithelial cell predominant type. Treatment of myasthenia gravis has been improved, however it is still unclear that how to treat invasive thymoma, and how to prevent re appearance of symptoms by decrease or discontinuity of PSL administration. PMID- 10791083 TI - [Brain and rhythm: from neurological aspect] [In Process Citation] PMID- 10791084 TI - [Pathogenetic mechanism of experimental autoimmune neuropathy induced by sensitization with a ganglioside]. AB - Antiganglioside antibodies are frequently present in sera from patients with autoimmune neuropathies. To elucidate the pathogenetic mechanisms of autoimmune neuropathies mediated by antiganglioside antibodies, we established a rabbit model of sensory ataxic neuropathy induced by sensitization with ganglioside GD1b (GD1b-SAN). Degeneration of primary sensory neurons extending the central axons to the dorsal column of spinal cord was observed pathologically. No lymphocytic cell infiltration was seen. Anti-GD1b antibody therefore should be an essential factor to induce GD1b-SAN. In sera from rabbits immunized with GD1b, two types of antibodies were present; antibodies monospecific to GD 1 b and those cross reactive with GM1. Of 22 rabbits sensitized with GD1b, 12 developed GD1b-SAN. The level of IgG antibody monospecific to GD1b was higher in the sera from affected rabbits than in those from unaffected ones. The GD1b-positive neuronal cytoplasms of rabbit dorsal root ganglia had larger diameters than the GD1b-negative ones. Markedly reduced expression of trkC in dorsal root ganglia from rabbits with GD1b SAN in acute phase was found. IgG antibody monospecific to GD1b may cause GD1b SAN by preferentially binding to large primary sensory neurons mediating proprioceptive sensation. Anti-GD1b antibody-mediated downregulation of trkC expression could be one of the pathogenesis of GD 1 b-SAN. PMID- 10791085 TI - [Treatment of anti-glycolipid antibody-associated neuropathy]. AB - The therapeutic effect of plasma exchange in Guillain-Barre syndrome (GBS) is related to the removal of circulating factors. Plasma concentrations of interleukin and other cytokines are elevated in GBS, but because their circulating half-lives are only a few hours, the effect of plasma exchange on their plasma levels would be short term. In contrast, the half-life of IgG is 3 weeks, much longer than that of other plasma proteins. This indicates that IgG is the most important factor in the development of GBS. Results of a large clinical study suggest that 2 exchanges are sufficient for mild cases of GBS, and for all others 4 exchanges are optimal. We showed that a significant IgG decrease was obtained in the first 2 sessions. Our result supports that at least 2 exchanges are needed for treating GBS. In Japan, patients with GBS often receive not only plasma exchange but also double-filtration plasmapheresis or immunoadsorption. We compared their abilities to remove immunoglobulins and anti-ganglioside antibodies. Plasma exchange decreased the IgG concentration much more than double filtration plasmapheresis. The ability to remove IgG anti-ganglioside antibodies in plasma exchange was significantly superior to that of the other plasmaphereses. PMID- 10791086 TI - [Antibodies directed to voltage-gated potassium channels in sera from acquired neuromyotonia and related disorders]. AB - Anti-voltage-gated potassium channel (VGKC) antibodies were measured in sera from 21 patients with acquired neuromyotonia (Isaacs syndrome) and related disorders. Seventeen of 21 sera suppressed potassium currents of neuroblastoma cell line (NB 1) using the patch clamp method, and the finding is mostly correlated with myokymic discharge in electromyography. Regarding the pathophysiology of suppression of VGKC, we investigated the effects of sera from 2 ANM patients on channel kinetics using the patch clamp method. The activation and inactivation kinetics of the potassium current were not altered by patients' immunoglobulins. We also investigated the effect of patients' sera on different VGKC subtypes using hKv1.1 and 1.6 transfected CHO cells. The same patients' sera suppressed the expressed VGKCs. Our results suggest that anti-VGKC antibodies may be positive not only in patients with classical ANM (Isaacs syndrome) but also in patients with related disorders who did not fulfill the characteristic symptoms. These disorders may be categorized as potassium channelopathy of the peripheral nervous system. The pathophysiology of the suppression of VGKC by antibodies is not the direct effect on channel kinetic but due to increased degradation or reduced production of VGKC. Epitopes of Anti-VGKC antibodies are still unclear. PMID- 10791087 TI - [The Lambert-Eaton myasthenic syndrome: a study of 110 Japanese cases]. AB - This study was undertaken to analyse the clinical and immunological features in a large group of Lambert-Eaton Myasthenic Syndrome (LEMS) patients (n = 110). In the Japanese LEMS patients studied, there was a male predominance with a male to female ratio of 3:1. The age at onset of neurological symptoms ranged between 17 and 80 years with a mean of 62 years. Malignancy was detected in 69% of the patients, of whom 61% had small cell lung carcinoma (SCLC). Neurological symptoms preceded a diagnosis of malignancy in 84% of cases. The neurological findings were similar in all patients and consisted of lower limb weakness in 97%, upper limb weakness in 80%, hyporeflexia in 85%, autonomic dysfunction in 37% (dry mouth in 31%, constipation in 11%, impaired sweating, urinary disturbance, impotence, and blurred vision in less than 10%), blepharoptosis in 28% and ophthalmoplegia in 5%. Signs of cerebellar involvement are noted in 9% and all of these patients had SCLC. Of 110 patients with LEMS, 85% had detectable antibodies against P/Q-type voltage-gated calcium channel (P/Q-type VGCC). Seronegative patients (15%) had similar neurological findings, but a lower incidence of SCLC than seropositive patients. The clinical features of our patients were very similar to those observed in British LEMS patients (n = 50), but autonomic features in our study were less prevalent than reported in British patients. PMID- 10791088 TI - [Nitric oxide-induced neurotoxicity versus neuroprotection; relationship with selective motor neuronal death]. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by selective motor neuronal death. In addition to elucidate the "cell death mechanism", we think it is also important to clarify the "cell survival mechanism", to understand the pathogenesis of this intractable disease. Glutamate (Glu) is an excitatory neurotransmitter in the central nervous system, and is implicated in the pathogenesis of ALS. In this report, we presented our current research, investigating the mechanism of Glu-induced selective motor neuronal death, derived from the study of primary culture of rat embryonic spinal cord. In brief, 1) motor neurons are selectively injured by long-term exposure to low-dose Glu through the activation of nNOS to generate NO and ONOO-: 2) nonmotor neurons are protected by cGMP which is formed by NONdependent guanylyl cyclase: 3) chronic exposure of spinal neurons to Glu increases nNOS positive neurons only in nonmotor neurons. These results indicate the cascade of Glu-calcium influx-NO generation is toxic to motor neurons and protective to nonmotor neurons. The different effect of cGMP on motor neurons and nonmotor neurons against Glu induced excitotoxicity may explain the selective motor neuronal death of ALS. Further investigation might advance the possibility of new therapy against ALS. PMID- 10791089 TI - [A motor unit number estimate (MUNE)--a quantitative and pathophysiologic parameter for amyotrophic lateral sclerosis (ALS)]. AB - We performed rate studies to develop a new method to obtain a MUNE reflecting the number of spinal alpha-motor neurons (MN). In the physiological part, six unitary muscle action potential (uMAPs) of the medial gastrocnemius muscle (MG) were averaged to obtain the mean uMAP, and a MUNE was calculated by dividing the area of its compound muscle action potential (CMAP) by the area of the mean uMAP. In the anatomical part, cholera toxin was injected into the MG muscle, and we counted the number of MG MNs identified by immunohistochemical staining of the toxin in their somata. We found that a MUNE of the MG muscle was consistent with the number of MG MNs. In the clinical application, we obtained MUNEs in 10 patients with ALS and 20 control subjects. In each patient we recorded 10 uMAPs and CMAP at each of the hypothenar and extensor digitorum brevis muscles using fine needle electrodes placed in these nerves. MUNEs for these muscles were calculated in the same way as described above. In the ALS patient group at one year post-onset of symptoms, the MUNEs for these two muscles were decreased to about 30% of the norm. We concluded that progression of ALS could be quantified by MUNEs. PMID- 10791090 TI - [Magnetic resonance axonography (MRX)]. AB - Neuronal fibers, especially axonal fibers, have empirically been implicated in the generation of the observed anisotropism in apparent diffusion within the nervous system. Fast axonal flow is believed to play a main, if not sole, role in producing this observed anisotropy. Given that apparent diffusivity follows a Gaussian distribution, "averaged" diffusion length may be calculated as a function of diffusion coefficient (eigenvalue) and diffusion time. This method can be developed into a non-invasive imaging technique for obtaining an estimate of fast axonal flow in humans, in vivo. In this lecture, one of such technique, magnetic resonance axonography (MRX) is briefly described. PMID- 10791091 TI - [Cotreatment of amyotrophic lateral sclerosis patients]. AB - In the treatment of cancers and infectious diseases, several drugs are administered simultaneously. Riluzole is a only drug which prolongs survival in amyotrophic lateral sclerosis (ALS) patients, however its effect is modest. We review the preclinical data supporting combination treatment for ALS and discuss the possible combination treatment in patients with ALS. In vitro studies showed favorable results in combination of neurotrophic factors. The combination of BDNF and GDNF reduced motor neuron death after axotomy for more than either factor alone. CNTF and BDNF combination treatment in wobbler mice arrested paw deformity. In the treatment of ALS, riluzole is the only drug available by prescription. When combination treatment is considered in ALS, the most effective combination need to be addressed. PMID- 10791092 TI - [Parkin gene and its function; a key to understand nigral degeneration]. AB - In most patients with Parkinson's disease (PD), the contribution of genetic factors as well as environmental factors remains to be elucidated. But, it has become clear that genetic factors contribute to the pathogenesis of PD after identification of the distinct genetic loci for certain forms of familial PD. We recently identified the novel large gene "parkin" responsible for an autosomal recessive form of familial parkinsonism (AR-JP). AR-JP is a distinct clinical and genetic entity characterized by early onset before 40 years. Pathological changes in this form revealed selective degeneration of the pigmented neurons in the substantia nigra and locus coeruleus, but no Lewy bodies were found. The parkin gene encodes a novel protein of 465 amino acids. The parkin gene is mildly homologous to ubiquitin at the N-terminal portion and has a RING-finger motif at the C-terminal portion. We found variable different homozygous deletions involving exons 3, 4, 5, 3 to 4, 3 to 5, and 3 to 7 in AR-JP families from Japan. In addition to exonic deletions, we identified a one base deletion in exon 5 in two AR-JP families. Although we have identified several mutations in parkin gene, characterization of its gene product, "Parkin protein" has not yet been established. To elucidate the molecular mechanism underlying the disease, we have analyzed the subcellular localization of the Parkin protein by immunohistochemical and immunoblotting studies on patients with AR-JP and sporadic PD using two antibodies. Parkin protein was absent in all regions of the brains of AR-JP patients. Parkin protein was not decreased in brains of sporadic PD patients. Parkin protein was located in both Golgi complex and cytosol. Taken together, the Parkin protein may play a role in vesicular transport system in association with the Golgi complex. PMID- 10791093 TI - [Familial parkinsonism and dementia with ballooned neurons, argyrophilic neuronal inclusions, atypical neurofibrillary tangles, tau-negative astrocytic fibrillary tangles, and Lewy bodies]. AB - We reported a new type of familial Parkinson's disease (familial PD) previously. After that, we examined this family by both the immunohistochemical staining using anti-alpha-synuclein antibody and the analysis of alpha-synuclein gene. We reported these results, and briefly reviewed both familial PD and "familial frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17)". Immunohistochemical staining with anti-alpha-synuclein antibody revealed that the argyrophilic neuronal inclusions and atypical barely tau-positive neurofibrillary tangles were strongly immunoreactive, whereas the ballooned neurons and astrocytic fibrillary tangles were unreactive. DNA analysis of the leukocytes obtained from one live patient of our family did not show any mutations in the entire exons of alpha-synuclein gene. Our results indicated that our family had familial diffuse Lewy body disease with atypical features and without alpha synuclein gene abnormalities. Features of familial PD included 1) autosomal dominant inheritance, 2) not uncommon atypical clinical features, 3) variable symptomatology and dopa-responsiveness, and 4) low incidence of alpha-synuclein gene abnormalities. Our familial PD showed similar features, and neuropathological findings of our patients also resembled FTPD-17 in the presence of frequent ballooned neurons and neurofibrillary tangles in the cerebral cortex, but were different in the presence of Lewy bodies and paucity of tau pathology. PMID- 10791094 TI - [Therapeutic effect of repetitive transcranial magnetic stimulation in Parkinson's disease]. AB - The therapeutic effect of repetitive transcranial magnetic stimulation (r-TMS) on clinical performance was studied in 8 patients with Parkinson's disease (PD). Seven patients were used as controls and underwent sham stimulation. The modified Hoehn and Yahr (H & Y) Staging Scale, Schwab and England Activities of Daily Living (ADL) Scale and Unified Parkinson's Disease Rating Scale (UPDRS) were used to assess changes in clinical performance. Eight patients were assessed prior to and following 3, 6 and 9 months of R-TMS. R-TMS was applied manually 60 times (30 times each side) to the frontal areas using a large circular coil, a pulse intensity of 700 V, and a frequency of 0.2 Hz. Sessions were performed once weekly for 9 months. The 7 control patients showed no differences in clinical symptoms between initial evaluations and evaluations after 3 months of sham R TMS. In all 8 patients, the modified H & Y staging and UPDRS scores decreased significantly, and the Schwab and England ADL Scale increased significantly after 3, 6 and 9 months of R-TMS therapy. These results suggest that R-TMS is beneficial for the treatment of Parkinsonian symptoms. PMID- 10791095 TI - [Positional cloning of the gene for Miyoshi myopathy and limb-girdle muscular dystrophy]. AB - Miyoshi myopathy (MM) is autosomal recessive distal muscular dystrophy that we have mapped to chromosome 2 p13. We constructed a 3 Mb P 1-derived artificial chromosome contig spanning the MM candidate region. Using this and new polymorphic markers within it, we recently identified a novel, full-length 6.9 kb muscle cDNA, whose corresponding protein we designated "dysferlin" (Nature Genet, 1998: 20: 31-36). We described eighteen mutations in the dysferlin gene with MM or limb-girdle muscular dystrophy type 2 B (LGMD 2 B). Most are predicted to block translation of dysferlin protein. In some cases, corresponding Western immunoblotting reveals absence of dysferlin in muscle biopsies. Identical mutations in the dysferlin gene can produce more than one myopathy phenotype (MM, limb-girdle dystrophy, distal myopathy with anterior tibial onset). PMID- 10791096 TI - [Sarcoglycanopathy]. AB - Sarcoglycanopathy is a group of four autosomal recessive muscular dystrophies whose symptoms are similar to Duchenne muscular dystrophy (DMD). These dystrophies are caused by mutations on anyone of the genes encoding four subunits of sarcoglycan complex which are transmembranous and dystrophin associated proteins. When the protein product of the mutated gene is absent, entire sarcoglycan complex is absent or greatly reduced in amount. This further gives rise to the weak connection between dystrophin and dystroglycan complex. These cause Duchenne-like phenotype. In DMD, dystrophin is absent and sarcoglycan complex is greatly reduced. These similarities in molecular defects in these diseases may cause the similarity in symptoms. PMID- 10791097 TI - [Formation of neuronal inclusions and neurodegeneration: with special reference to dentatorubral-pallidoluysian atrophy]. AB - A new entity--the conformational disease--an otherwise diverse group of disorders, includes several currently incurable brain diseases such as Alzheimer's disease, prion encephalopathies and inherited polyglutamine diseases (PGDs) in which it is proposed that conformational changes in certain proteins and their subsequent aggregation represent the common pathogenic mechanism of neurodegeneration. In several PGDs including dentatorubral-pallidoluysian atrophy (DRPLA), neuronal intranuclear inclusions (NIIs) have been identified. Here I describe our recent studies on DRPLA. Formation of both intranuclear and intracytoplasmic inclusions by mutant DRPLA proteins with an expanded polyglutamine stretch was demonstrated in an in vitro experiment. Inclusion containing non-brain cells were found to undergo apoptosis. In human DRPLA, similar intranuclear inclusions were detected in neurons in various brain regions. However, intracytoplasmic inclusions were found exclusively in neurons in the cerebeller dentate nucleus. The pontine nuclei neurons with NIIs, for instance, appeared to be small (atrophic), but healthy. The question of whether these NIIs actually kill brain cells in the human disease, remains unclear. Recently, NIIs were studied immunohistochemically using antibodies against transcription factors, and were found to contain TAFIIp130, Sp 1 and CREB-1. These results are of great interest when considering not only how the neurodegenerative mechanism progresses but also why the inclusions are located in nuclei in PGDs. Finally, Tsuji and colleagues recently established a transgenic mouse model of DRPLA. In these mice, NIIs, which were identical to those seen in the human disease, were confirmed in various brain regions. Further detailed studies of this model will contribute to a better understanding of the formation of neuronal inclusions and neurodegeneration in DRPLA. PMID- 10791098 TI - [Familial amyotrophic lateral sclerosis associated with mutant Cu/Zn superoxide dismutase as a conformational disease]. AB - To investigate the molecular mechanism of mutant Cu/Zn superoxide dismutase (SOD) associated with familial amyotrophic lateral sclerosis (FALS), mutant (A1a4Thr, Gly85Arg, Gly93Ala, and two base-pair deletion in the 126th codon), as well as wild-type (wt), Cu/Zn SODs were expressed in COS7 cells. The formation of granular cytoplasmic aggregates accompanied by collapse of the cytoplasm was observed in cells expressing mutant Cu/Zn SODs, but not in cells expressing mutant Cu/Zn SODs. The aggregates contained ribosome-like particles and endoplasmic reticulum. These results suggest the possibility that mutant Cu/Zn SODs promote the formation of aggregates which are toxic to cells. PMID- 10791099 TI - [Parkinson's disease, dementia with Lewy bodies, multiple system atrophy and alpha-synuclein]. AB - Lewy bodies (LBs) are hallmark lesions of degenerating neurons in the brains of patients with Parkinson's disease (PD) and dementia with Lewy bodies (DLB). DLB is the second most common neurodegenerative dementia after Alzheimer's disease, which is characterized clinically by fluctuating cognitive impairments, visual hallucinations and parkinsonism, and pathologically by the appearance of cortical LBs. To characterize the components of LBs, we have developed a purification procedure for LBs from cortices of patients with DLB using sucrose density separation followed by fluorescence-activated particle sorting. We then raised monoclonal antibodies (mAbs) to purified LBs, and obtained a mAb (LB509) that intensely immunolabeled LBs and specifically reacted with a approximately 18kDa brain protein, which was identified as alpha-synuclein. LB509 as well as other antibodies to alpha-synuclein, but not to beta-synuclein, immunostained brainstem and cortical LBs in sporadic PD and DLB brains. Recently, a point mutation in alpha-synuclein gene was identified in some autosomal-deminantly inherited familial PD pedigrees. Moreover, glial cytoplasmic inclusions in the brains of patients with multiple system atrophy (MSA) were shown to be alpha-synuclein positive. Taken together, our data strongly implicate alpha-synuclein in the formation of LBs and the selective neuronal degeneration in PD, DLB and MSA. PMID- 10791100 TI - [Microcirculatory derangement and apoptosis in ischemia-reperfusion injury]. AB - This study was undertaken to clarify the involvement of microcirculatory failure and apoptosis in the pathophysiology of cerebral ischemia using ICAM-1 knockout mice (K/O) and BCL-2 transgenic mice, respectively. In both permanent and transient focal ischemia, infarcted size of cerebral cortex in ICAM-1 K/O mice was significantly smaller than that in wild type mice. Microcirucaltaory disturbance in the cerebral cortex after permanent and transient focal ischemia was mitigated in ICAM-1 K/O mice compared with that in wild type mice. However, the number of granulocytes in the infarcted tissue was similar between K/O and wild mice, and neutrophil depletion in K/O mice showed further reduction of cortical infarction after transient focal ischemia. In contrast, neuronal overexpression of BCL-2 in mice showed protective effect on selective neuronal death observed in the hippocampus after transient global forebrain ischemia for 12 min. The present study supported the notion that (1) microcirculatory disturbance mediated through interaction of ICAM-1 and leukocytes played an important role in expansion of cerebral infarction after focal ischemia and (2) apoptosis inhibited in part by overexpression of BCL-2 was involved in selective neuronal vulnerability after transient global ischemia. PMID- 10791102 TI - [Alteration of cAMP-mediated signal transduction in cerebral ischemia--binding activity of PKA and phosphorylation of CREB]. AB - Binding of cAMP to the regulatory subunit of cAMP-dependent protein kinase (PKA) is an essential step for cAMP-mediated signal transduction including phosphorylation of cAMP response element binding protein (CREB). In the present study, binding activity of PKA with cAMP and CREB phosphorylation were examined in rat focal brain ischemia induced by occlusion of the middle cerebral artery for 1.5 hours followed by various time of recirculation. Binding activity of PKA with cAMP was progressively inhibited during the acute phase of ischemia from the ischemic core to peri-ischemia area. Phosphorylated CREB-positive cells in the ischemic core revealed a significant, but transient increase in number at 3.5 hours of recirculation, followed by a rapid decrease below the control level during the subsequent period. On the other hand, in the peri-ischemia area, the number of phosphorylated CREB-positive cells showed a more marked increase as compared to that in the ischemic core, and the increase continued until 48 hours of recirculation with a tendency for gradual decline. Persistent enhancement of CREB phosphorylation may thus be closely related to the neuronal viability and neuroprotective mechanisms, whereas rapid disappearance of CREB phosphorylation following ischemic insult may clearly precede neuronal death. PMID- 10791101 TI - [Peroxynitrite production in cerebral ischemia]. AB - Peroxynitrite, generated by the reaction of nitric oxide and superoxide, has toxic effects including oxidation of sulfhydryls, lipid peroxidation and nitration of amino acid residues. So far peroxynitrite has not yet been detected in the ischemic brain because of its short half-life. Recently, we have succeeded in detecting 3-nitro-L-tyrosine, which is considered to be a footprint of peroxynitrite, in ischemic brain. Production of nitrotyrosine started during the ischemic period, increased after reperfusion, peaked at 48 hours, then declined up to 72 hours. Nitrotyrosine level was highest in the peri-infarct region, second highest in the core-of-infarct region, and lowest in the caudoputamen and the non-infarct region. Studies using pharmacological agents including MK-801, 7 nitroindazole and aminoguanidine suggest that peroxynitrite production originates from nNOS in the early phase of reperfusion, and from iNOS in the later phase of reperfusion. Further, the immunohistochemical study indicates that iNOS, located mainly in vascular cells, is predominantly responsible for nitrotyrosine production. Thus, peroxynitrite production depends on the stage of evolution of the ischemic process and on the cell type producing NO. These findings have important implications for the therapeutic time window and choice of NOS inhibitors in patients with cerebral infarction. PMID- 10791103 TI - [Studies on brain pyruvate dehydrogenase (PDH) activity and energy metabolites during ischemia and reperfusion]. AB - Pyruvate dehydrogenase is one of the mitochondrial enzymes considered important in the regulation of oxidative metabolism. To further understand the relationship between its activity and ischemic brain damage we conducted three experiments. We studied the effects of (1) duration of cerebral ischemia, (2) the Ca2+ channel blocker, nicardipine, and (3) the immunosuppressant, FK506, on PDH activity and energy metabolites during ischemia and reperfusion. In the first study we also measured regional cerebral blood flow (rCBF). (1) Increasing the duration of the ischemic insult delayed the deactivation of PDH, slowed the resynthesis of high energy phosphates and the clearance of lactate, and impaired recovery of rCBF. Additionally, (2) nicardipine normalized PDH activities and improved the impaired metabolism after reperfusion, and (3) FK506 did not effect PDH activity, but significantly improved the impaired metabolism during the early phase of reperfusion. From these studies we conclude that PDH plays a role in the recovery of metabolism during reperfusion, and both nicardipine and FK506 improve metabolism during the early phase of reperfusion. PMID- 10791104 TI - Monitoring ambient air quality for health impact assessment. PMID- 10791106 TI - Hyperthyroidism exacerbating asthma. PMID- 10791105 TI - Hyperthyroidism complicating asthma treatment. AB - Asthma is one of the most common chronic medical conditions. The usual treatment includes quick relief bronchodilator medications of the sympathomimetic class and controller medications that may include the long-acting inhaled bronchodilator salmeterol. Mild adverse cardiac and central nervous system effects are common with these medications, requiring modifications in dose or occasionally switching to a different medication. Both asthma and thyroid disease are common disorders that occasionally occur together. Hyperthyroidism may exacerbate asthma. Many symptoms of hyperthyroidism are identical to the adverse effects of the commonly used inhaled bronchodilators and include tremor, nervousness, tachycardia, wide pulse pressure, palpitations, emotional lability, agitation, nightmares, aggressive behavior, and diarrhea. In this report we describe a patient with hyperthyroidism whose symptoms initially were thought to be adverse effects of the inhaled bronchodilator medications. PMID- 10791107 TI - Dual-phase response model for bronchial asthma. AB - The authors have successfully developed an animal model of dual-phase bronchial responses and very high IgG titer by sensitizing Hartley-strain male guinea pigs. Specific airway resistance, which was determined in a two-chamber body plethysmograph, was elevated to sevenfold during immediate response, followed by a late phase response with a smaller but marked elevation in resistance. Furthermore, hematological and histological examinations revealed that the total cell count increased in BAL obtained during both immediate and late bronchial responses as compared to pre-OVA challenges. A significant increase in BAL eosinophils was present only for the late bronchial samples, and this finding was supported by histological examination. PMID- 10791108 TI - The risk of sinusitis in children with allergic rhinitis. AB - It is said that bacterial sinusitis occurs often in patients with allergic rhinitis, but this correlation has not yet been determined. We studied this issue for 5 years in 413 children, of whom 215 had perennial allergic rhinitis (PAR) and 198 had seasonal allergic rhinitis (SAR). The prevalence of sinusitis in patients with PAR compared with those with SAR was determined with respect to the following factors: age, season, and sensitivity to airborne allergens. We found that the prevalence of sinusitis was significantly higher among patients with PAR than among those with SAR regardless of age or season. When the risk of sinusitis was compared between patients with PAR and SAR, patients with mold allergy PAR had a higher risk than those with non-mold allergy (PAR relative risk 2.49 versus 1.50, respectively). We conclude that mold allergy is an important risk factor for sinusitis, and that its management may help to reduce the occurrence of sinusitis. PMID- 10791110 TI - ECP level in nasopharyngeal secretions and serum from children with respiratory virus infections and asthmatic children. AB - Infection with respiratory virus has been shown to exacerbate asthma in humans. However, the role of a respiratory virus in the pathogenesis of chronic asthma and/or wheezing in young children has not been clearly defined. It has also been debated whether virus-induced wheezing in young children is one entity and allergic asthma another, or whether they are different expressions of the same disease. The present study was done to compare ECP concentrations in nasopharyngeal secretions and serum from 32 nonasthmatic wheezing children with viral infections (RSV in 15 children; influenza B virus in 17 children detected by immunofluorescence antibody technique), 8 asthmatic children without viral infections, and 13 normal children as the controls to understand the role of eosinophil inflammation. The geometric mean of ECP in nasopharyngeal secretions was significantly higher in asthmatic children than in children with virus induced wheezing (p < 0.05). ECP levels of nasopharyngeal secretions from children with the virus-induced wheezing were significantly greater than those of the controls. However, there were no significant differences in ECP levels in serum among subjects. PMID- 10791109 TI - Increase of the soluble IL-4 receptor (IL-4sR) and positive correlation between IL-4sR and IgE in nasal fluids from school children with allergic rhinitis. AB - Soluble cytokine receptors (SCR) can either act as inhibitors, by competitively inhibiting cytokines from binding to their membrane-bound receptors, or as enhancers, by serving as cytokine carriers. We have previously found that the levels of the Th2 cytokines interleukin (IL)-4, IL-5, IL-6, and IL-10 were positively correlated to eosinophils and IgE in nasal fluids from 60 children with seasonal allergic rhinitis. In this study, nasal fluids were reexamined to analyze IL-4sR, IL-6sR, IL-1 beta, TNF-alpha, IL-1sR2, TNF-sR1, and TNFsR2 in relation to eosinophils, neutrophils, ECP, and IgE. In allergic patients IL-4sR increased significantly during the pollen season, and weak, but positive correlations with IgE and eosinophils were found (r = 0.45, P < 0.001 and r = 0.4, P < 0.001 respectively). By contrast, none of the other SCR showed increases or correlations with IgE. However, positive correlations between IL1 beta, TNF alpha, IL-6sR, IL-1sR2, TNF-sR1, TNF-sR2, and either neutrophils or ECP were found. Also, in healthy controls, these cytokines and their receptors were positively correlated to neutrophils or ECP. Thus, increased levels of the soluble IL-4 receptor, as well as IgE, were specifically associated with allergic rhinitis, whereas all other SCR correlated with either inflammatory cells or their products, in both allergic and healthy subjects. These results may suggest that SCR in vivo act as cytokine enhancers, rather than inhibitors. PMID- 10791111 TI - Corticosteroid therapy in an additional 13 cases of Stevens-Johnson syndrome: a total series of 67 cases. AB - Stevens-Johnson syndrome (SJS) is a severe cutaneous eruption that can be a life threatening emergency. Previously, we have reported our favorable experience in treating 54 patients with SJS with systemic corticosteroids. We continued our prospective analysis of consecutive patients with SJS treated with corticosteroids. Possible etiologic factors and clinical outcomes of the patients are described. All 13 patients improved with initiation of systemic corticosteroid therapy. There was no mortality or permanent sequelae attributable to SJS. Drugs were the offending agents in all 13 cases. There was one death unrelated to SJS. In conclusion, prompt treatment with systemic corticosteroids reduces morbidity and improves outcome of SJS patients. This analysis extends our series to 67 consecutive patients with SJS who were treated with corticosteroids and had a favorable outcome. PMID- 10791113 TI - Venom immunotherapy: when to start, when to stop. AB - Over the past 25 years, major advances have been made in the diagnosis and treatment of insect sting allergy. Controlled clinical trials have demonstrated the efficacy of venom immunotherapy (VIT) in the prevention of subsequent systemic reactions in allergic individuals. We have refined our criteria for selection of patients for VIT. Studies on selections of venoms, rush immunotherapy, and interval between VIT injections have been performed. Finally, much work has been done to try to define criteria for the discontinuation of VIT. PMID- 10791112 TI - The role of atypical organisms in asthma. AB - Atypical organisms (Chlamydia pneumoniae, Mycoplasma pneumoniae) have been recently linked to asthma in various ways: an infection with these organisms may precede asthma onset, exacerbate asthma, or make asthma control more difficult. Their ability to elicit a TH2 response and promote airway inflammation may be the common pathway in the development of an atopic inflammatory response. This article presents a summary of the evidence that infection with Chlamydia pneumoniae or Mycoplasma pneumoniae may play a significance role in asthma. PMID- 10791114 TI - From immunity to autoimmune disease, a historic trail. Part II. PMID- 10791115 TI - Vicarious liability: relocating responsibility for the quality of medical care. PMID- 10791116 TI - Iceland's Health Sector Database: a significant head start in the search for the biological grail or an irreversible error? PMID- 10791117 TI - First nursing homes, next managed care? Limiting liability in quality of care cases under the False Claims Act. PMID- 10791118 TI - The Patients' Bill of Rights: women's rights under managed care and ERISA preemption. PMID- 10791119 TI - A review on melioidosis with special respect on molecular and immunological diagnostic techniques. AB - Melioidosis is an 'emerging' tropical disease which causes diagnostic problems in endemic and especially in nonendemic areas e.g. Germany when imported. In the last decade, many efforts have been made to develop new molecular and immunological techniques for diagnostic use. However, an actual comprehensive review does not exist. Apart from classical microbiological procedures (microscopy, culture and biochemical identification) efforts have been made to identify Burkholderia pseudomallei using specific antibodies and PCR. The direct antigen detection can be done within a few hours leading to diagnosis days before cultural proof. ELISA and immunoblot techniques were examined for their ability to replace indirect hemagglutination and immunofluorescence test in serology. The diagnostic value of serological procedures for early detection of melioidosis is limited, however. The rapid and reliable diagnosis of melioidosis is required for an adequate onset of therapy. But no evaluated test kit based on the detection of specific antibodies, specific antigens, or on the amplification of species specific DNA sequences is commercially available up to now. Even PCR testing- primers can easily be ordered by many gene technology companies--can not be recommended as B. pseudomallei DNA for positive controls is not available. Therefore, only microscopy and biochemical identification systems like the API 20NE can be used in the routine laboratory up to now. At this point, it has to be stressed that B. pseudomallei is a level 3 agent in many countries e.g. Germany and that only laboratories with high containment are allowed to handle it. In all cases the help of an experienced reference laboratory is adviced. PMID- 10791120 TI - Evaluation of a sensitive immunoluminometric assay for the determination of C reactive protein (CRP) in serum and plasma and the establishment of reference ranges for different groups of subjects. AB - A sensitive immunoluminometric assay originally designed to measure C-reactive protein (CRP) in neonates and minimal serum volumes was adapted to measure this protein in a routine method without prior sample dilution. The concentration range covered without prior dilution was 10 micrograms/l to 20 mg/l using a sample volume of 5 microliters serum and a total assay time of less than 2 h. Serum samples were assayed from participants in a community medicine programme (SHIP--Study of Health in Pomerania) of the University of Greifswald, Germany (n = 414), as well as from mother-child pairs at birth (n = 30) and women attending the infertility clinic (n = 36). The validation of the assay was compared with a commercial latex-enhanced turbidimetric immunoassay (Roche Diagnostics--Integra 700) using routine serum samples (n = 60) from hospital patients. Comparison was made with the routine assay used in the SHIP study (Roche Diagnostics--Hitachi 717/Tina Quant). From 414 SHIP samples measured in the immunoluminometric assay, 289 were below the detection level in the turbidimetric (Tina Quant) assay. A significant positive correlation (p < 0.01) between log C-reactive protein concentration with age was found, both in the non-screened (all CRP values) (n = 414, r = 0.222) and selected (CRP < 5.00 mg/l = 90th percentile) (n = 370, r = 0.242) SHIP participants. Women were found to have significantly higher CRP levels than men (women: median age 47 a, median CRP 1.29 mg/l; men: median age 55 a, median CRP 1.00 mg/l--p = 0.016) in the non-selected SHIP participants. The situation was different in the selected group, (median age: men 54 a, women 48 a) where no significant difference in median CRP values between the sexes was seen (men: 0.874 mg/l, women 0.951 mg/l, p = 0.206). The distribution of CRP values in a "Normal Healthy Population" is skewed (mean/median--SHIP: all--2.08; selected- 1.49). From the 414 SHIP samples measured in the immunoluminometric assay, 289 were below the detection level (2.5 mg/l) in the turbidimetric (Tina Quant) assay. From the 125 remaining samples the correlation between both methods was acceptable (r = 0.813), the regression line y = a + bx being: CRP (ILMA) = 1.83 + 0.842*CRP (Tina Quant). The Tina Quant assay gave values significantly higher than the ILMA in the range 2.5-25 mg/l CRP (p < 0.001). The total information loss in 289/414 subjects with a CRP < 2.5 mg/l with the Tina Quant assay makes it no longer suitable for epidemiological studies in which CRP is to be studied as a risk factor for cardiovascular events. The comparison between the immunoluminometric assay and the latex-enhanced immunoturbidimetric assay (Roche Integra) was much better. The latter measured down to less than 0.3 mg/l, thus being more suitable for epidemiological studies than the Tina Quant assay from the same producer. The correlation and regression data between the ILMA (x) and the Roche Integra assay (y) were: r = 0.971; CRP (Roche Integra) = 0.635 + 0.984*CRP (ILMA); n = 50.10 sera with CRP levels between 25 and 460 mg/l showed no high-dose hook effect in either assay. The remaining 50 sera were measurable in both assays. The turbidimetric assay gave rise to marginally but significantly higher values than the immunoluminometric assay (p = 0.004). The mothers at birth had a median CRP of 3.64 mg/l (range 1.49-12.6 mg/l), the neonates a median CRP of 34 micrograms/l (range 4-288 micrograms/l). All births were without complications, with gestational periods between 38 and 42 weeks. There was no correlation between maternal and neonatal CRP at birth. Mothers at birth had significantly higher CRP levels than healthy non-pregnant women (p < 0.001). Women attending the infertility clinic had CRP-values similar to age-matched healthy non-pregnant women (median 0.698 mg/l, range 0.05-9.97 mg/l). Interassay coefficients of variation at CRP concentrations of 0.85 and 7.9 mg/l were 8.99 and 7.93%, respectively, for the immunoluminometric PMID- 10791121 TI - Homocysteine in renal disease. PMID- 10791122 TI - Lipids and stability of menopausal status in middle-aged women within two years. AB - In the whole population total cholesterol (TC) was unchanged within two years. The triglyceride (TG) level increased significantly, but HDLC increased too between 1996 and 1998. The increase of TG concentration was significantly in the stable group only, whereas the increase of HDLC concentration was observed in both groups. "Depressed mood" and "anxiety" seem to have a positive influence on the TC levels. PMID- 10791123 TI - Evaluation of a centrifuge with rapid turnaround time for the preparation of plasma samples for measurement of common STAT markers on the ACS: 180 system. AB - Reported is the evaluation of a new centrifugation method, Statspin, that addresses both time and sample separation integrity. The method can successfully separate the plasma fraction from the cellular material in 2 minutes as compared to 20 minutes for the conventional centrifuge method. The Statspin, combined with the ACS:180 system, can generate test results in less than 30 minutes, exclusive of transport to the laboratory. This study demonstrated that the combined technologies offer timing-saving improvements for clinical laboratories offering STAT immunoassays for cardiac markers, endocrine molecules, and therapeutic drugs. PMID- 10791124 TI - The detection of recombinant hepatitis B surface antigen from "vaccine escape mutants" in two HBsAg immunoassays. PMID- 10791125 TI - Development of a novel ELISA for 1,25-dihydroxyvitamin D. PMID- 10791126 TI - External evaluation of LIAISON tumour marker assays on the fully automated chemiluminescent LIAISON immunoassay analyser. AB - The LIAISON immunoassay analyser was tested in a multicentre evaluation performed by 8 laboratories. The analytes evaluated were CA 15-3, CA 19-9, CA 125II, AFP, CEA, NSE and PSA. Excellent results were obtained for within-run and between-run precision with most assays showing within-run CVs < 5% and between-run CVs between 4 and 8%. The linearity of all assays was acceptable, however, for PSA, NSE and CA 19-9 a recovery > 110% was obtained for some of the samples tested. None of the assays revealed a high-dose hook effect. Method comparisons were performed by using the routine method of the respective study centre. Results generally showed an acceptable agreement between the LIAISON system and the different methods of comparison. The reference ranges for all assays were found to be in accordance with data known from the literature. All assays showed similar results for serum, heparinised plasma and EDTA plasma. Additionally, two experiments were performed with only one of the analytes tested: the sample-to sample carry-over, using the CA 19-9 assay (3.3 x 10(-6)-2.3 x 10(-5)) and the functional sensitivity for the PSA assay (0.2 ng/ml). PMID- 10791128 TI - Molecular risk factors for thrombosis and risk factors in venous thrombotic disease. PMID- 10791127 TI - Evaluation of the LIAISON thyroid chemiluminescence immunoassays. AB - The LIAISON thyroid hormone assays TSH, FT4, FT3, T4 and T3 were evaluated by determining the imprecision, the reference ranges, the functional sensitivity (TSH), the dilution characteristics (accuracy) (FT4, FT3), and the recovery after spiking (TSH, T4, T3). Furthermore, inter-method comparisons were performed with following methods: Elecsys (Roche Diagnostics; TSH), AxSYM (Abbott Diagnostics; TSH, FT4, FT3, T4), ACS:180 (Bayer Diagnostics; all analytes), Amerlex-M (Johnson & Johnson; T4) and LISO-Phase (Techno Genetics; FT4). The fully automated LIAISON random access analyser is based on microparticle immunoassays and chemiluminescence. The coefficients of variation (CV) of intra-assay imprecision were between 0.2-6.0%, except for the control sample with extremely low TSH concentrations and low T3 concentrations. Inter-assay imprecision was performed by measuring controls covering the measuring range over a period of 9 to 20 days, with CVs ranging from 2.3-16.0%. The suitability of the sample material was determined by analysing serum and samples treated with EDTA, citrate or heparin in parallel. The results showed good correlations of the thyroid hormone concentrations between serum and plasma samples except for LIAISON FT3, for which lower results were observed with EDTA-plasma. The regression analysis of correlation studies gave slopes from 0.849 to 0.957 for TSH, from 1.023 to 1.375 for FT4, from 0.670 to 0.911 for FT3, from 0.917 to 1.166 for T4 and 1.00 for T3 depending on the concentration range and the method of comparison. The LIAISON FT4 assay showed a trend towards higher values in the high concentration range when compared with the ACS:180. The ranges of thyroid hormone concentrations determined in serum taken from apparently healthy subjects were found to be in accordance with published data. The clinical sample study confirmed that the LIAISON thyroid hormone assays are sensitive methods for the differentiation of euthyroid subjects and patients with hyper- and hypothyroidism. In conclusion, the automated thyroid hormone immunoassays on the random-access LIAISON immunoassay analyser proved to be very satisfactory, both from the analytical and the clinical point of view. PMID- 10791129 TI - Heparin-induced thrombocytopenia. PMID- 10791130 TI - Neurologic manifestations of Mycoplasma pneumoniae infections: diverse spectrum of diseases. A report of six cases and review of the literature. AB - Mycoplasma pneumoniae is a common cause of upper and lower respiratory tract infections of varying severity. It is also responsible for producing a wide spectrum of nonpulmonary manifestations including neurologic, hepatic, cardiac, and hematologic diseases. The neurologic manifestations are reported to be the most common nonpulmonary manifestations. We describe six patients demonstrating the protean neurologic manifestations of Mycoplasma pneumoniae infections. Four patients presented with the central nervous system manifestations of pyramidal and extrapyramidal tract dysfunction, seizures, cognitive abnormalities, and cerebellar dysfunction. Two patients presented with transverse myelitis. The outcome of this condition ranges from normal to severe residual deficits. Increased awareness of this disease entity may facilitate early diagnosis and thereby expedite starting appropriate therapy that may modify the outcome. PMID- 10791132 TI - Diagnosis and prospective treatment of enteroviral infections in children. PMID- 10791131 TI - Detection of enteroviruses in the cerebrospinal fluid by polymerase chain reaction: prospective study of impact on the management of hospitalized children. AB - A polymerase chain reaction kit (AMPLICOR EV) for the detection of enteroviruses (EV-PCR) in the cerebrospinal fluid (CSF) was evaluated in clinical conditions in a prospective blinded-intention study. Forty-three children (mean age 2.7 years) hospitalized for suspected meningitis or fever of unclear etiology were enrolled. EV-PCR was performed on a daily basis. Results were available in less than 2 days in 72% of cases. EV-PCR was positive in nine (21%) children, including three infants without CSF pleocytosis. Knowing their EV-PCR result would have allowed a saving of 18 hospital days and 12 days of antibiotic therapy. The EV-PCR in the CSF can thus be practically useful for children hospitalized for meningitis or fever if available on-site on a daily basis. PMID- 10791133 TI - Prednisolone plus albuterol versus albuterol alone in mild to moderate bronchiolitis. AB - To evaluate combination therapy of mild to moderate bronchiolitis with bronchiodilators and corticosteroids, we treated 51 young children with first time wheezing and symptoms of respiratory tract infection with albuterol plus either prednisolone or placebo for 5 days. Disease severity was scored on days 0, 2, 3, and 6. On day 2, prednisolone resulted in significantly lower scores (2.7 +/- 1.4 vs. 4.0 +/- 1.5 in all patients evaluated, p < 0.05) than placebo, whereas there was no detectable difference on day 6, suggesting that addition of prednisolone to albuterol transiently accelerates recovery from bronchiolitis. The clinical significance of this effect needs to be evaluated in further studies. PMID- 10791135 TI - The association of celiac disease, diabetes mellitus type 1, hypothyroidism, chronic liver disease, and selective IgA deficiency. PMID- 10791134 TI - Establishing a pediatric hospitalist program at an Academic Medical Center. AB - Academic medical centers have been challenged to respond to a rapidly changing and increasingly competitive health care environment. The Pediatric Consultation and Referral Service (PCRS) at Rainbow Babies & Children's Hospital (RB&C)/University Hospitals of Cleveland was established in 1993 with the goal of providing rapid access to community-based physicians for the referral of patients requiring urgent hospitalization within the broad scope of general pediatrics. We describe our initial 3-year experience in the planning, implementation, and evaluation of a pediatric hospitalist program. PCRS provided care to 2,740 patients during the first 3 years of operation, 63% (1,716) of whom were under age 3 years. Leading primary diagnoses in order of decreasing frequency were asthma, pneumonia, bronchiolitis, febrile illness, gastroenteritis, seizures, croup, apnea, and cellulitis. Third-party payer mix was: Medicaid 42%, managed care 42%, indemnity insurance 10%, self-pay 6%, and Bureau for Children with Medical Handicaps 1%. From survey data, referring physicians and pediatric residents assessed perceptions of access, collegiality, and quality of care in a highly favorable manner. Subspecialty colleagues perceived access and collegiality very favorably but rated quality of care substantially lower than referring physicians and residents did. Our experience demonstrates that a pediatric hospitalist program is logistically and economically feasible and may contribute to the patient care, education, and research missions of academic medical centers. A well-structured program can provide community physicians with excellent access and support collegial relationships. Beyond increasing a medical center's patient referral base, a hospitalist program can potentially enhance the esteem of the discipline of general pediatrics and, it is hoped, promote general pediatrics as a viable career option for trainees. PMID- 10791136 TI - Postnatal developmental delay and chromosomal abnormalities. PMID- 10791137 TI - Digital desquamation--a new finding in perianal streptococcal dermatitis. PMID- 10791138 TI - Sickle cell anemia: severe ischemic colitis responding to conservative management. PMID- 10791139 TI - Pediatric ibuprofen use increases while incidence of Reye's syndrome continues to decline. PMID- 10791140 TI - Prevalence and clinical outcome of hepatitis C infection in children who underwent cardiac surgery before the implementation of blood-donor screening. PMID- 10791141 TI - Interstitial nephritis in children with Crohn's disease. PMID- 10791142 TI - Unusual location of subcutaneous fat necrosis in a term newborn. PMID- 10791143 TI - Parietal whorl (crown): how many in Saudi children? PMID- 10791144 TI - Introduction. The magnitude of skin disease in the United States. AB - Skin disease is common in the United States and accounts for a considerable fraction of all outpatient visits. Dermatologists care for more skin disease visits than any other single specialty, while still accounting for a minority of visits overall. The data presented in this introduction, from 1997, do not show a drastic drop in the number of visits to dermatologists as might be feared to occur with the growth in managed care. These visits for skin disease represent a considerable disease burden in the United States. Skin disease is not simply a cosmetic problem. As the data on psoriasis show, skin disease has a tremendous impact on HRQL. The magnitude of this impact on the individual patient is comparable to the impact of other medical disorders. PMID- 10791145 TI - Dermatology in the developing world. Needs and solutions. AB - Dermatologists, who have assets far in excess of most of the people of the world, have the opportunity to contribute globally to the forces of social progress. This article addresses the dermatologic needs of the developing world and discusses how the needs of these areas, including rural areas in the United States, should be addressed. PMID- 10791146 TI - Economic aspect of health care systems. Advantage and disadvantage incentives in different systems. AB - European health care delivery systems illustrate the effect of economic incentives on health care delivery. Each country faces the issue of trying to balance the desire for economic efficiency with comprehensive, quality medical care. Without careful use of economic incentives achievable with central control, one gets to pick only two of the three desired goods--high quality, low cost, and comprehensive coverage. In the United States, payment approaches for health care have been undergoing tremendous changes since the early 1980s. These changes have escalated during the 1990s. The basic approach for reimbursing hospital care has been completely restructured by many payers for care, and payment approaches for physicians and long-term care providers also are being restructured. Financing approaches vary from provider to provider and payer to payer, and financing approaches will continue to evolve over time. In the traditional fee-for-service reimbursement system, the incentive to physicians is to do more because more services lead to more revenue. The use of incentives to influence health care practitioners' behavior is common. Incentives are generally financial in nature and expose health care providers to some risk or reward for certain patterns of behavior. Some common incentives used in managed care include capitation payment, in which a physician is paid a fixed fee, regardless of the number of services administered; bonus distribution; and withhold accounts, through which a practitioner stands to gain or lose some amount of money for overuse or underuse of medical resources against budget. In many countries, a strengthening of the position of primary care providers can be observed: Finland, Germany, Greece, Italy, the Netherlands, Norway, Sweden, the United Kingdom, and now the United States. General practitioners are assumed to function as a gatekeeper to second line care, such as specialist care, prescription drugs, and hospital care. A further step is to give the primary care providers financial responsibility for the costs of the follow-up care provided by others to their patients. By examining the health care systems of other countries, the potential negative impact of such an approach on the use of specialists can be seen. The negative impact of these approaches on patient care is clear when dermatologists and general practitioners are compared in the delivery of dermatologic health care. PMID- 10791147 TI - The primary care physician and the treatment of patients with skin disorders. AB - Scientific advances have altered the nature of primary care medicine. Primary care providers are increasingly asked to care for a broad spectrum of common maladies, including cutaneous disease. We review studies that have compared primary care physicians with dermatologists with respect to the diagnosis and treatment of skin disorders. Though primary care providers play a crucial role in the delivery of health care, the judicious and appropriate use of dermatology consultation should be encouraged. PMID- 10791148 TI - Role of the dermatologist in the delivery of dermatologic care. AB - Dermatology faces an uncertain future because of changes in the mechanisms of health care delivery. The Westwood-Squibb Center for Dermatology Research was created to foster the care of patients through research and education that supports the delivery of optimal dermatologic health care. The center performs health care research to effect policy decisions that impact dermatologic health care delivery. Results from a broad range of these studies clearly demonstrate the value of dermatologists as primary care providers for patients with skin disease. Future studies at the Westwood-Squibb Center for Dermatology Research expand on these findings, demonstrate the cost-effectiveness of care by a dermatologist, and address the expertise of dermatologists at management of surgical problems of the skin. PMID- 10791149 TI - Issues in dermatologic health care delivery in minority populations. AB - It is well known that African Americans are at low risk for one of the most serious diagnoses in dermatology--skin cancer. A consequence of this lower cancer risk has been an undervaluing of the importance of appropriate dermatologic care in this population. In fact, there are many other skin diseases and disorders that not only cause significant morbidity among this population but also emotional and psychological sequelae. This article examines barriers and challenges to providing dermatologic care among African Americans and provides recommendations and suggestions for improving care delivery in this population. PMID- 10791150 TI - The role of dermatologists in the delivery of occupational dermatologic care. AB - This article reviews the role of dermatologists in occupational dermatologic care by reviewing the types of skin diseases that may be work-related, the diagnostic testing, and evidence of quality of care delivery and outcomes provided by dermatologists. PMID- 10791151 TI - Delivery of dermatopathologic health care in the twenty-first century. AB - Dermatology and dermatopathology has experienced significant growth over the last century and continues to burgeon. Upheavals as a consequence of the advent of managed care have threatened both specialties, especially with regard to direct access and choice of consultant. The demand for dermatology and dermatopathology services continues to increase, and for patients to be served appropriately, these issues must be addressed. PMID- 10791152 TI - Issues in the delivery of dermatologic surgery for skin cancer. AB - Significant financial resources need to be dedicated to treating the large numbers of skin cancers in the United States. The goal of the surgical treatment of skin cancer is to provide a high cure rate in a manner that is minimally morbid, convenient, and inherently of high value. Cost-effective delivery of this skin cancer care is critical given the prevalence of cutaneous neoplasia. Dermatologists are the best-trained physicians in the identification and management of melanoma and NMSCs. They are the most highly skilled judges of when intervention is indicated, of which lesions warrant biopsy, and of which treatment techniques are applicable for each individual tumor. These skills take years to develop and are necessary to provide the highest standard of medical and surgical management of skin cancer. As dermatologic surgery continues to evolve and dermatologists in residency receive even more surgical training, the dermatologist will prove to be the surgical skin cancer specialist well into the new millennium. PMID- 10791153 TI - Evidence-based medicine in dermatology. AB - EBM is the use of the best current evidence in making decisions about the care of individual patients. Practicing EBM requires five steps: (1) formulating well built clinical questions, (2) finding the best evidence to answer the questions, (3) critically appraising the evidence, (4) applying the evidence to specific patients, and (5) saving the critically appraised evidence. The Cochrane Library is the best source for finding the best evidence about treatment. The second best method for finding evidence about treatment and the best source for finding most other types of best evidence in dermatology is by searching the MEDLINE database using the PubMed Clinical search engine of the National Library of Medicine (http://++www.ncbi.nlm.nih.gov/PubMed/clin ica l.html). MEDLINE searches have inherent software and operator limitations that make their reliability quite variable. The quality (strength) of evidence is based on a hierarchy of evidence: results of systematic reviews of well-designed clinical studies, results of one or more well-designed clinical studies, results of large case series, and expert opinion. Once the best evidence has been found, the EBM approach involves critically appraising the quality of the evidence, determining its magnitude and precision, and applying it to the specific patient. Guidelines to appraise critically and apply evidence are available. The clinical question, best evidence, and its critical appraisal should be saved in a format that can be easily retrieved for future use. PMID- 10791154 TI - Patient advocacy groups. A key prescription for dermatology. AB - In addition to medical care, patient education and support is important to successful treatment of chronic dermatologic diseases. For assistance, dermatologists and patients should turn to patient advocacy groups, which provide educational materials, programs, support groups, and advocacy for patients with chronic skin diseases. PMID- 10791155 TI - The roles of the pharmaceutical industry and drug development in dermatology and dermatologic health care. AB - Drug development is becoming shorter, more high-tech and strategic, more costly, and more complicated. The pharmaceutical, biotech, and cosmetic companies, along with regulatory agencies such as the FDA, are struggling to cope with and master the scientific, medical, and economic implications of this new environment. There are rapidly growing new classes of drugs, including biologicals, genomics, antibodies, and novel receptor-ligand antagonists. Dermatologic drug development has several idiosyncrasies, including the vehicle in topical drugs. Development for dermatology is much cheaper than for other therapeutic areas but also generates much less sales. The pharmaceutical industry's search for blockbusters threatens to leave dermatology without access to these new technologies, therapeutic modalities, and drug classes. The pharmaceutical industry is interested, invested, and intertwined, at many different levels, in the efforts and practices of academic dermatology, dermatology specialty organizations, and clinical dermatologists. PMID- 10791156 TI - The emerging role of physician assistants in the delivery of dermatologic health care. AB - The NAMCS provides a wealth of information on use of PAs in all practices, including dermatology. Two important points regarding the NAMCS and SDPA data are addressed here: the number of visits to PAs for dermatologic symptoms and the expected growth of PA use in dermatologists' offices. Dermatologic symptoms were evaluated frequently by PAs, accounting for 14% of PA visits. These statistics do not address the number of referrals those PAs made to dermatologists. Perhaps PAs as a group should be targeted for increased dermatologic education, particularly stressing the need for appropriate referral to a dermatologist. PAs could increase the number of dermatology referrals from primary care offices with improved understanding of the importance of the dermatologist in the management of patients' overall skin health. At projected growth rates, the number of PAs employed by dermatologists should exceed 500 by the end of 2000. Most of this growth has been in private practices and rarely in HMOs or in large multispecialty clinics. There are a number of reasons for this growth, as follows: A PA may help reduce the patient load on the dermatologist, especially with sameday appointments and drop-ins. Some dermatologists are moving away from clinical dermatology into cosmetics, which not only leaves a vacuum in clinical dermatology, but also creates job opportunities for PAs in cosmetic dermatology. Regarding managed care growth, PAs can have a positive impact on the problem of having to see more patients for less money. PAs are cost-effective. In the 1998 SDPA survey, the ratio of billings generated (production) to gross income for the average dermatology PA ranged from 3:1 to 6:1. Even with inexperienced PAs new to dermatology, this ratio was usually at least 2:1 at the end of the first year. PAs can cover satellite offices, allowing for practice expansion. Effective with the new Medicare laws of January 1, 1998, PAs can now see new Medicare patients or Medicare patients with new conditions without the physician being on site, opening up the possibility for satellite offices in remote areas. Just as dermatologists may move toward specialization in surgery, cosmetics, or medical dermatology, PAs may do the same, filling a niche in a particular practice. As in other specialties, patient acceptance of seeing dermatology PAs has not been a significant problem. Continued access to the dermatologist remains unfettered, but, over time, many patients become willing to see either. Are PAs likely to become future competitors of dermatologists? Genuinely concerned dermatologists worry that a dermatology-trained PA will become part of a gatekeeper system that impedes patient access to dermatologists. This is not happening and is not at all likely to become a trend, for a number of reasons. First, primary care cannot compete with dermatology practices in remuneration for PAs. Just as financial benefits in high-production specialty practices entice physicians, the same benefits entice PAs as well. Second, according to member surveys of the SDPA, virtually 100% of fellow members work with dermatologists. Although PAs can work in any type of practice and evaluate dermatologic symptoms just as a general practitioner would, PAs who specialize in dermatology primarily practice with dermatologists, a collegial association most PAs seek out. PAs have steadfastly maintained their dependent, noncompetitive relationship with physicians and would not have it any other way. Although PAs see a good number of patients (2.8 million) with dermatologic symptoms, the NAMCS data indicate that most (72%) of these patients are also seen by a physician. Third, physicians are ultimately responsible for the actions of their PA employee. A general practitioner not trained to perform excisions or manage certain dermatologic conditions should not allow a PA to perform such duties. Similar to much of medicine, the PA profession continues to evolve, with many members moving awa PMID- 10791157 TI - Dermatology manpower needs. AB - The future need for dermatologists is difficult to predict, as it depends on the structure of our health care delivery system, which is still rapidly evolving, and whose future shape is unknown. The best strategy to deal with this unpredictable situation is to de-emphasize trying to predict the future and focus on responding as rapidly as possible to whatever changes in dermatologic manpower needs the future may bring. From that perspective, it is best to train fewer rather than more dermatologists, as it is much quicker to rectify a shortage than an excess in medical manpower. Other issues that impact on dermatologist manpower include a very rapid increase in nonphysicians taking care of skin diseases, the maldistribution of dermatologists, the decreasing proportion of common skin diseases treated by dermatologists, and the need to upgrade the residency training to master the explosion in medical knowledge which has occurred, and to gain a greater expertise in the medical and cosmetic aspects of the specialty. PMID- 10791158 TI - Growing impediments to the delivery of dermatologic care. AB - Government regulations have a broad and deep impact on the delivery of dermatologic health services. It is incumbent on dermatologists to be involved in the development and discussions of these issues and to educate government officials about the effects of existing and proposed regulations on the delivery of dermatologic health care. PMID- 10791159 TI - Organizing state dermatologic societies. AB - Each year state legislatures, boards of medicine, departments of insurance, and other authorities promulgate laws and rules that affect the daily practice of dermatology. It is critical that dermatologists in each state form and maintain a strong state dermatologic society. Specific suggestions for forming and strengthening such societies are described and available resources are reviewed. PMID- 10791160 TI - Unusual, innovative, and long-forgotten remedies. AB - A compendium of uncommon dermatologic treatments, both medical and surgical, are presented. This article is based on a survey of the members of the Noah Worcester Dermatologic Society with subsequent additions, including treatment pearls from the Internet and other contributions from practicing dermatologists, as well as some published material. Most of the therapies discussed in this article are based on anecdotal information. PMID- 10791161 TI - Treatment of common nail disorders. AB - Treatment of nail diseases is not always easy; topical treatment is often ineffective because the nail plate is not penetrated by topical drugs, and the nail matrix is located under the proximal nail fold. The most frequent nail disorders are nail psoriasis and onychomycosis. In these cases, systemic treatment is often necessary to cure the nail changes. Systemic treatment with steroids is mandatory for patients with nail lichen planus because the disease may cause definitive destruction of the nail matrix. Environmental nail abnormalities are frequent and include nail brittleness, onycholysis, and chronic paronychia. Preventive measures should always be associated with treatment in these cases. PMID- 10791162 TI - Establishing a practical and effective psoriasis treatment center. AB - Running an effective psoriasis treatment center requires 1) capable phototherapists who have received the appropriate education and training; 2) a full range of equipment, including light boxes, hand-foot units, scalp debridement machines, and a whirlpool bath; 3) adequate space; 4) economic considerations, including reimbursements so patients can receive treatment and the center can be paid; and 5) the medical expertise necessary to care for the most severe psoriatic patients. These and other issues are discussed in this article. PMID- 10791163 TI - Nontuberculous mycobacterial infections of the skin. AB - Nontuberculous mycobacteria are playing an increasingly important role in human disease owing to higher prevalence of antibiotic resistance and immunodeficiency. These organisms cause a variety of cutaneous findings which are often misdiagnosed by the clinician. Compounding this problem is the fact that most mycobacteria require special culture conditions, which if not specifically requested, are frequently not used. Recognition of susceptible patients is imperative and is not limited to the immunocompromised. Successful treatment of mycobacterial infections requires knowledge of currently available and recommended antibiotics followed by tailoring of the antimicrobial regimen after sensitivity testing is performed. PMID- 10791164 TI - Escharotomy and fasciotomy of the burned upper extremity. AB - Escharotomy and fasciotomy are performed in the burned upper extremity to prevent and treat the sequelae of circumferential full-thickness burns and high-voltage electrical burns. Indications to perform these procedures are determined primarily by clinical examination but can be supplemented by measurements of subfascial pressures. The techniques are designed to avoid further complications. Options for wound closure are discussed. The use of allograft as temporary coverage of fasciotomy incisions may allow delayed primary closure. PMID- 10791165 TI - Management of acute burn injuries of the upper extremity in the pediatric population. AB - Burns of the upper extremity occur frequently in children. Because of differences in development and anatomy, patterns of burn injury are different in children compared to adults. Immediate goals after these injuries are to prevent compartment syndromes and minimize progressive damage. The second decision is whether the burn requires conservative care or grafting. If the injury heals within 2 weeks, then scarring is minimized. If the wound has not healed in that time period, then grafting should be considered. Grafting techniques that optimize function and cosmetic appearance are outlined. PMID- 10791166 TI - Management of the acutely burned upper extremity. AB - In summary, thermal injury is a frequent from of upper extremity trauma that often affects young working-class males. Its management must be designed with preserving life as a priority, recognizing the additional risk factor of inhalation injury as well as the traditional predictors of mortality--age and proportion of the TBSA burned. The aim of care in dorsal hand burns is to restore wound coverage and movement within 14 days of injury. Determining the depth of thermal injury to the upper extremity is notoriously difficult and requires serial assessment over time to most accurately predict the time of healing, still considered the standard measure of burn depth despite newly developing technical assessment tools. Tangential excision and split-thickness autograft coverage are predictably uniformly successful for deep dermal burns in restoring function in both the short and long term; however, burns exposing underlying tendons, bones, or joints require individualized reconstructive procedures, including local and distant flaps, free-tissue transfers, or combinations of these procedures. Early TAM is the most successful modality for recovery of function and to minimize any one of many potential complications associated most commonly with deep wounds and prolonged periods of healing. PMID- 10791168 TI - Management of chemical injuries to the upper extremity. AB - Chemical burns are interesting and challenging to treat. When the practitioner is comfortable with the pathophysiology of chemical injury and treatment based on these principles, most patients can be treated effectively with good outcome (Fig. 2). Early treatment with water irrigation, followed by diligent wound care, will allow our patients to benefit to the maximum from our medical skills. PMID- 10791167 TI - Modern techniques for wound coverage of the thermally injured upper extremity. AB - The use of flaps; synthetic, dermal analogues; cultured skin substitutes; or a combination of these may one day help return severely burned arms to normal function and appearance. The complexity and expense of these alternatives limits their use in commonplace burns. As the techniques are refined with increased experience, so also will the roles for each of these options. Fortunately, an excellent result is still possible in most situations using simple autograft. The availability of these numerous choices to acutely cover the thermally injured upper extremity should not distract us from the gold standard of split-thickness skin graft (Fig. 7). PMID- 10791169 TI - Upper extremity electrical injury. AB - Electrical injuries to the upper extremity are far ranging in extent and vary in magnitude. Proper management requires an appreciation for the pathophysiology, clinical manifestations, and therapeutic options. Persistent neurologic and psychiatric problems further impact the high incidence of disability in the electrical injury patient. Loss is significant for victims, their families, and employers, in part because of the relative youth of those injured. PMID- 10791170 TI - Frostbite of the upper extremity. AB - Human capacity for physiologic adaptation to cold is minimal; we survive by insulating ourselves with protective clothing. In addition to the irreversible direct injury caused by ice crystallization, the authors have outlined four possible mechanisms by which indirect injury may damage tissue. Other than rapid rewarming, there is no uniformly accepted protocol for the treatment of frostbite injury. Attempting to sort out the world's literature on frostbite in an effort to present a comprehensive treatment protocol is a daunting task. In addition to the probably irreversible direct injury caused by ice crystallization, the authors have outlined at least four possible mechanisms by which indirect injury may damage tissue. The literature is full of various treatment protocols that allegedly are beneficial despite addressing different mechanisms. Mills described 10 different categories of medications, each addressing one of four possible mechanisms, used in the clinical treatment of frostbite injury over a 30-year period. Analyzing this information is even more confusing when one realizes that there is little uniformity in animal models employed to generate these data. This is further complicated by the lack of clinical correlation with the most common experimental model--liquid nitrogen rapid freezing. The risk of frostbite is highest when psychiatric disturbance, intoxication, or unplanned circumstances lead to cold exposure without adequate protective clothing. As tissue freezes, both direct and indirect factors cause injury. Most therapies have been aimed at limiting indirect injury, in an attempt to limit progressive tissue loss. Rapid rewarming is universally accepted, but the benefits of other modalities are still controversial. Traditionally, observation and delayed amputation have been employed to manage frostbite. More recently, triple-phase bone scans have been used to distinguish between tissue that is irreversibly destined for necrosis and tissue that is at-risk for necrosis, but potentially salvageable. Early operation can be used to provide at-risk tissue with a new blood supply and preserve both function and length in the upper extremity. PMID- 10791171 TI - Reconstruction of the pediatric burned hand. AB - It is clear that reconstruction of pediatric hand burns is a complicated task (Fig. 8), and attention to details during the acute phase of injury may be surgeons' greatest ally in subsequent functional rehabilitation of the hand. Reducing edema, maintaining digital circulation, limiting inflammation, and mobilizing the limb early are key parameters to assure return of function during the acute phase of injury. Although children may present with neglected contractures at a later date, even grade IV contractures can be surgically approached with hopes of improving hand function. Loss of the thumb in children with very large total body surface area burns can be approached reliably by lengthening of the thumb using first-to-second metacarpal transfers, as described by Littler. Although electrical injuries represent only a small fraction of patients admitted to our hospitals, children who present with viable insensate hands can have reliable sensory return more than 1 year after injury using nonvascularized cable nerve grafts if soft tissue coverage is adequate. As surgical procedures continue to evolve in delayed reconstruction of the hand, one would expect children to show better results than those seen in adult patients. A more aggressive surgical approach, with increased optimism, therefore is required when addressing children with complex hand burns requiring reconstruction. PMID- 10791172 TI - Microsurgical reconstruction of the thermally injured upper extremity. AB - The use of free flaps for coverage of the deeply burned hand has advantages that include the salvage of the exposed vessels, nerves, tendons, joints, and bone; a single operation to obtain wound closure, minimizing the risk of infection; and earlier physical therapy. This article focuses on the choice of suitable free flaps for the coverage of the deeply burned hand; and it also presents some case reports. PMID- 10791173 TI - Molecular and cellular aspects of fibrosis following thermal injury. AB - The pathogenesis of hypertrophic scars following thermal injury remains a complex and incompletely understood process but recent investigations into the composition of the tissue itself, the activities of the scar fibroblasts, and the effects of various cytokines and growth factors, have all contributed to the emergence of an increasingly clear picture. Although it may be considered just one example of a broad range of fibroproliferative disorders that afflict many different organs, often in response to diverse environmental insults, the nature of the burn injury and the special properties of skin probably play important roles in promoting the development of this especially troublesome variety of excessive connective tissue. This knowledge has provided the rationale for a number of experimental therapies that, individually or in some combination, may augment or one day supplant the more commonly employed surgical or physical treatments. PMID- 10791175 TI - Rehabilitation of the burned upper extremity. AB - With the advancement in medical technology and more effective life-sustaining measures, the rehabilitation therapist is faced with the immense task of effectively restoring functional ROM, strength, and mobility and producing a cosmetic result acceptable to the patient. Rehabilitation therapists have a very significant role to play in achieving these goals. The patients and their families come to rely very heavily on the therapists for advice, support, and information both in the acute phase of burn management and, potentially, for years to follow. A concerted team approach is necessary for a satisfactory functional outcome following burn injury. PMID- 10791174 TI - Control of wound contraction. Basic and clinical features. AB - Although a substantial amount of molecular and cellular data have been generated in an effort to understand the process of wound contraction and scar contracture formation, questions remain. What seems apparent is that the myofibroblast is not the only cell that generates contractile forces within wounds, but it does appear to be intrinsically linked to the development of hypertrophic scars. The supposition that the formation of scar contractures is solely the result of a continuation of wound contraction is an oversimplification. Figure 4 provides a model of the possible evolution of contractile forces during the wound healing process and their role in the development of scar contractures. Migration of fibroblasts into and through the extracellular matrix during the initial phase of wound healing, prior to the expression of alpha-SMA, appears to be a fundamental component of wound contraction. During this migration, the pulling of collagen fibrils into a streamlined pattern in their wake, and the associated production of collagenase, may facilitate a more normal arrangement of collagen. Once the wound has been repopulated and the chemotactic gradient that was established by inflammatory cells is decreased, fibroblast migration will cease. It is at this point that myofibroblasts appear and play a key role in the production of hypertrophic scars, given that their prolonged presence and over-representation are hallmarks of this pathology. One of the pivotal differences between wounds that proceed to normal scar compared with those that develop hypertrophic scars and scar contractures may be a lack (or late induction) of myofibroblast apoptotic cell death. The combined contribution of fibroblasts and myofibroblasts to abnormal extracellular matrix protein production results in an excessive and rigid scar. The isometric application of contractile forces by myofibroblasts probably contributes to the formation of the whorls, nodules, and scar contractures characteristic of hypertrophic scars. Because the prolonged presence of myofibroblasts, producing an imbalance in extracellular matrix proteins and proteases, probably exacerbates hypertrophic scars and wound contraction, accelerating the rate of apoptotic cell death to reduce the cell number to that seen in normal scar may be a useful strategy for providing effective and efficient treatment of scar contracture. PMID- 10791176 TI - Rectal cancer. PMID- 10791177 TI - Screening for rectal cancer. AB - Although screening for rectal cancer remains a controversial topic, the case for screening has been strengthened by the demonstration that screening can reduce mortality from the disease. Three randomized controlled trials based on fecal occult blood testing have shown a significant reduction in overall colorectal cancer mortality in those offered testing. The case for screening based on flexible sigmoidoscopy is not as strong. Evidence from randomized controlled trials will not be available for another 8-10 years. Whatever screening method is chosen, factors such as the level of participation and the cost-effectiveness of protocols for adenoma follow-up will have major effects on the success of screening. PMID- 10791178 TI - Prognostic factors in colorectal cancer. AB - Prognostic factors should always be taken into consideration before planning treatment. As reported in the literature, and also in our experience, the extent of intestinal wall invasion and the lymph node status must be considered the most important prognostic factors. In the present paper the data from literature are reported and the impact of biologic factors is analyzed. We are convinced that in the future the biological factors of the tumor as well as of the patient should be considered in planning treatment. PMID- 10791179 TI - Genetics of colorectal cancer. AB - The multistep nature of cancer has been well-documented through molecular genetic studies of colorectal cancer. The information provided by basic, translational and clinical science highlighted several genes associated with different hereditary syndromes predisposing to colorectal cancer. Due to these and other findings, the molecular pathogenesis of this cancer has been clarified also with respect to genetic pathways which can lead to an otherwise undistinguishable disease. As a result of both the available and continuously added data, our knowledge and understanding of colorectal cancer is subject to constant changes. Ultimately, these are expected to result in multiple and strategical managements based on molecular genetics which will likely change our approach to this cancer in the incoming millennium. PMID- 10791180 TI - Results of coloanal anastomosis for rectal cancer. AB - It is now accepted that, in the absence of direct invasion of the anal sphincter, cancers of the middle and lower rectum can be successfully treated with sphincter preserving surgery. Conservation of the sphincter mechanism should never compromise the oncologic outcome of surgery and the method of neorectum construction must provide acceptable function for patients. This review describes the results of coloanal anastomosis following rectal excision. The oncological and functional results of both straight coloanal and colonic-J-pouch anal anastomosis are presented in detail. The authors discuss recent evidence supporting the functional superiority of colonic-J-pouch reconstruction after rectal excision. PMID- 10791181 TI - Local resection of rectal cancer. AB - Stringent patient selection and careful preoperative evaluation are important factors for the successful treatment of all rectal cancers. The lack of histopathologic evaluation is the parameter that even in the best scenario, allows for a 5% risk of nodal involvement that cannot be assessed and therefore treated. However, in appropriately selected situations, local treatment of rectal cancer allows for effective and safe curative therapy. PMID- 10791182 TI - Which type of follow-up? AB - Most patients and many surgeons assume that proactive follow-up after rectal cancer surgery is useful in most cases and therefore an essential part of surgical care. The evidence base for this contention is sparse. What evidence there is suggests that there is little effect on the supervivability of recurrent cancer however intense, costly--or the opposite--the follow-up program may be. This review examines some of the issues in this difficult area, and suggests some reasonable conclusions on what degree of follow-up might be applied in the absence of a wide and easily applicable evidence base. PMID- 10791184 TI - Laparoscopic versus open cholecystectomy: effect on pulmonary function tests. AB - BACKGROUND/AIMS: Operations often cause impairment in respiration due to pain. This study was designed to compare the changes in pulmonary function tests after open and laparoscopic cholecystectomy. METHODOLOGY: Two groups of 35 patients were randomly set up. Each patient had 3 pulmonary function tests performed and 2 postero-anterior grid chest roentgenograms taken. All of these data were evaluated by the same group of investigators. RESULTS: After taking into consideration the difference between pulmonary function tests, values were not significant (P < or = 0.05). All pulmonary function test values decreased significantly on the 1st postoperative day (P < or = 0.05). When postero-anterior chest roentgenograms were compared no clinically evident atelectasis except 3 lineary was seen in the laparoscopic cholecystectomy group, whereas 5 lineary, 7 focal, and 3 segmentary atelectasia were encountered in the open cholecystectomy group (P < or = 0.05). CONCLUSIONS: We believe that laparoscopic cholecystectomy has more advantages when speaking of postoperative pulmonary function tests and atelectasia. PMID- 10791183 TI - Malignancy in choledochal cysts. AB - BACKGROUND/AIMS: Malignancy in choledochal cysts is a rare condition. This study presents our experience with this condition, with emphasis on the clinical presentation, management and outcome. METHODOLOGY: Subjects included 80 adults with choledochal cysts treated from January 1979 to December 1995. Of these patients, 8 were found to have malignancy in the cyst and formed the basis of this study. RESULTS: Four patients had synchronous and 4 had metachronous carcinoma lesions arising in the choledochal cyst. The clinical presentation was: biliary tract infection in 5 patients, gastric outlet obstruction in 2 and right upper quadrant pain and body weight loss in 1. Operations for bile duct malignancy included total excision in 2 patients, choledochotomy with T-tube drainage in 2 patients, gastrojejunostomy in 2 patients, percutaneous transhepatic biliary drainage and gastrojejunostomy in 1 patient and metastatic lymph node biopsy only in 1. One patient died due to septic shock within 30 days of the operation (operative mortality). Postoperative survival time ranged from 4 13 months with a mean of 6.2 months. CONCLUSIONS: The frequency of malignancy in the choledochal cysts was 10% in the present series. Malignancy in the choledochal cyst should be highly suspected in patients with cholangitis symptoms, body weight loss and anemia. Prognosis in this disease entity is poor. PMID- 10791185 TI - Relation between the risk of gallstone pancreatitis and characteristics of gallstone in Korea. AB - BACKGROUND/AIMS: Pancreatitis is a serious complication of patients with gallstones. However, risk factors of gallstone pancreatitis were unpredictable until recently. In Korea, characteristics of gallstones are different from Western countries. The present study was designed to determine differences in the risk of gallstone pancreatitis and characteristics of gallstones in Korean patients. METHODOLOGY: Clinical data were collected on patients undergoing laparoscopic cholecystectomy. The physical characteristics of gallstones recovered at surgery were also recorded. Patients with gallstone pancreatitis were compared with patients who had uncomplicated biliary pain. RESULTS: In a logistic regression model, acute gallstone pancreatitis was associated with a stone diameter of less than 5 mm (odds ratio: 3.3695; P = 0.0352) and with stone number of more than 20 (odds ratio: 3.8686; P = 0.0361). No other variable, including pigment stone, age, and sex, remained statistically significant in the adjusted analysis (P > 0.05). CONCLUSIONS: Patients with at least 1 gallstone smaller than 5 mm in diameter and stone number more than 20 each have a more than 3-fold increased risk of presenting with acute gallstone pancreatitis. The composition of gallstones, especially pigment stones, was not an important risk factor in gallstone pancreatitis in Korean patients with stones having a different composition than those from Western countries. PMID- 10791186 TI - Effect of omeprazole on gallbladder contraction in humans. AB - BACKGROUND/AIMS: Omeprazole causes hypergastrinemia because of the effects of prolonged complete suppression of acid secretion and also gastrin has an excitatory effect on gallbladder contraction. Therefore, we investigated the meal induced gallbladder emptying in healthy subjects receiving omeprazole and compared them to controls. METHODOLOGY: Twenty healthy volunteers participated in this study. Gallbladder volume was measured by ultrasonography. After basal measurement, the volunteers received saline intravenously (i.v.) 2 cc (no:10) or omeprazole 20 mg i.v. (no:10). After 15 min the gallbladder volume was scanned at 15 min intervals for 60 min for each of the subjects. At the end of the period, all the subjects received a standard test meal (ensure 250 cal/250 mL), after 1 hour the gallbladder volumes were rescanned at 15 min periods for 60 min. RESULTS: Mean gallbladder volume in the omeprazole group was not significantly different during a 45 min period as compared to the baseline value. The residual gallbladder volume at the end of the 15th minute (43.9 +/- 5.6 mL), 30th minute (45.4 +/- 5.9 mL), 45th minute (40.5 +/- 6.1 mL) and 60th minute (40.5 +/- 6.1 mL) showed no significant differences in both the omeprazole group and the controls. Mean gallbladder volumes of both groups after meal intake were significantly lower during the 1-hour period as compared to the baseline value (P < 0.05). The mean volumes did not show any significant differences between the omeprazole group and the control subjects. CONCLUSIONS: Omeprazole did not change the gallbladder volume during fasting and the postprandial period as compared to the control group. PMID- 10791187 TI - Extrahepatic bile duct carcinoma: a western experience with 118 consecutive patients. AB - BACKGROUND/AIMS: This study was designed to evaluate the effectiveness of a diagnostic and therapeutic approach adopted in 118 consecutive patients affected by primary malignancy of the extrahepatic bile duct. METHODOLOGY: After diagnostic procedures were performed (ultrasound examination, endoscopic retrograde cholangiopancreatography and computed tomography scan) 25 patients underwent surgical resection. For the remaining 93 patients considered unresectable for cure, stenting by endoscopic means was almost always the only palliative treatment performed. RESULTS: The morbidity and mortality rates were 28% and 8% for patients treated with surgical resection and the curative and overall 3-year survival rate was 30% and 22% respectively. The procedure related morbidity and mortality rates were 13% and 4% for patients endoscopically treated and the median survival rate was 7.3 months. The quality of life evaluated in 68 patients out of 93 was good in 57% of cases, fair in 19% and poor in the remaining 22%. CONCLUSIONS: The results of the present study demonstrate the safety and efficacy of endoscopic stenting in the palliative treatment of extrahepatic bile duct cancer while potential cure can only be achieved by radical surgical resection. PMID- 10791188 TI - Effects of vitamin E on neutrophil phagocytosis during experimental obstructive jaundice. AB - BACKGROUNDS/AIMS: Depression of non-specific immunity is one of the systemic complications of biliary obstruction. Vitamin E, which decreases during prolonged obstructive jaundice, may be beneficial to diseased function of neutrophils. In this study we want to investigate changes in neutrophil phagocytosis and the effect that vitamin E supplementation has on this function METHODOLOGY: Rats were divided into 5 groups as follows: the control group and 4 other groups that underwent double ligation and division of the common bile duct. Two of these 4 groups (Group 3 and 5) received vitamin E during the experiment. Alkaline phosphatase, aspartate aminotransferase, bilirubin serum levels, white blood cell count and neutrophil phagocytosis index were determined for group 2 and 3 at the end of the 15 days and for group 1, 4 and 5 at the end of the 21 days. RESULTS: There was a significant increase in white blood cell counts and biochemical parameters in group 2, 3, 4, and 5 (P < 0.05). Neutrophil phagocytosis index significantly increased 15 days after bile duct ligation (P < 0.001) (Group 2) and significantly decreased 21 days after bile duct ligation (P < 0.001) (Group 4). Neutrophil phagocytosis index in vitamin E pretreated groups were significantly decreased at the end of the 15 days (P < 0.001) (Group 3) and increased at the end of the 21 days (P < 0.001) (Group 5). CONCLUSIONS: Finally, If vitamin E is administered for further days and weeks of prolonged jaundiced, neutrophil phagocytosis index improves. PMID- 10791189 TI - Hypereosinophilic sclerosing cholangitis: findings using half-Fourier magnetic resonance imaging. AB - Hypereosinophilic sclerosing cholangitis is a rare disease caused by eosinophilic infiltration of the gallbladder and biliary tract seen in the idiopathic hypereosinophilic syndrome. We report a 42-year-old woman who presented with symptoms of cholecystitis and obstructive cholangitis. Imaging with magnetic resonance cholangiography using a half-Fourier spinecho sequence, we were able to visualize rapidly and non-invasively a severely abnormal gallbladder, evidence of liver parenchymal inflammation, and biliary duct dilatation. PMID- 10791190 TI - Resection of a hilar cholangiocarcinoma in a patient with absent portal bifurcation. AB - Absence of the bifurcation of the portal vein is an extremely rare anomaly and the misled ligation of the portal vein would result in the lethal hepatic failure. In this paper, a resected case of hilar cholangiocarcinoma with this anomalous portal venous system is first presented. Preoperative percutaneous transhepatic portography disclosed an absence of portal bifurcation and a transversely running portal vessel from the right anterior segment to the left lateral segment. The patient underwent left hepatic lobectomy with caudate lobectomy, in which the portal trunk was successfully preserved. PMID- 10791191 TI - Modulation of apoptosis, tumorigenesity and metastatic potential with antisense H ras oligodeoxynucleotides in a high metastatic tumor model of hepatoma: LCI-D20. AB - BACKGROUND/AIMS: To investigate the effect of antisense H-ras DNA on tumorigenesity, apoptosis and metastasis of a high metastatic tumor model of human hepatocellular carcinoma in nude mice LCI-D20. METHODOLOGY: LCI-D20 cells in primary culture were treated with 10 microns/L antisense oligodeoxynucleotide (ODN) drugs in vitro. 1.5 x 10(6) LCI-D20 cells with or without pretreatment were inoculated into each elevated subcutaneous (s.c.) flap in 14 nude mice, 6 animals for antisense H-ras oligodeoxynucleotide treated cells, 4 for H-ras non-specific antisense oligodeoxynucleotide treated cells, and the rest 4 for cells without pretreatment. RESULTS: In in vitro cell culture study, 5-day continuous suppression of H-ras expression by antisense H-ras oligodeoxynucleotide resulted in significant inhibition of the proliferation of LCI-D20 cells (t = 31.529, P < 0.01). In situ end-labeling detection showed that apoptotic cell death was significantly increased in cells with 5-day treatment of antisense H-ras oligodeoxynucleotide (34.0 +/- 4.5%) in comparing with cells without treatment (2.5 +/- 1.2%, t = 13.434, P < 0.01) or treated with non-specific antisense oligodeoxynucleotide (4.8 +/- 1.4%, t = 12.453, P < 0.01) at the corresponding time. In the in vivo experiment, at week 6, no palpable tumor could be found in 50% (3/6) of animals receiving cells with pretreatment of antisense H-ras oligodeoxynucleotide, while 100% (4/4, 4/4) of animals in the 2 control groups developed palpable tumors. Tumor growth in antisense H-ras treated animals was significantly retarded in comparison with that of the untreated (t = 3.509, P < 0.01) or non-specific antisense oligodeoxynucleotide treated animals (t = 3.452, P < 0.01). 75% to 100% of animals in the 2 control groups developed lung metastases, while in antisense H-ras treated animals lung metastasis foci could not be found by random serial section and microscopy (u = 2.536, P < 0.01; u = 3.162, P < 0.01, respectively). CONCLUSIONS: Specific inhibition of H-ras expression by antisense H-ras oligodeoxynucleotides could not only induce apoptotic cell death, inhibit the growth rate of LCI-D20 cells in vitro and in vivo, but also alter in vivo tumorigenesity and metastatic potential of LCI-D20 cells. PMID- 10791192 TI - Systematic analysis of the effects of hepatocyte transplantation on rats with acute liver failure. AB - BACKGROUND/AIMS: Acute liver failure either after liver resection or as part of underlying liver disease is still associated with high mortality. Hepatocyte transplantation in various forms has attracted attention recently. However, none of those reports have investigated the thorough and systematic analysis of effect of hepatocyte transplantation on acute liver failure induced by 90% hepatectomy. Therefore, we investigated systematic analysis of effect of hepatocyte transplantation on rats with acute liver failure. METHODOLOGY: Male Sprague Dawley rats were used. Group I rats (n = 26) received intrasplenic injection of 2 x 10(7) hepatocytes in 0.3 mL Dulbecco's modified Eagle's medium (DMEM) 24 hours prior to 90% hepatectomy. Group II rats (n = 24) received intrasplenic injection of DMEM only. Twenty-two rats from group I and 20 from group II were observed for the determination of survival time. The remaining 8 (4/each group) rats were used to assess the liver function and regeneration. RESULTS: The hepatocyte bearing spleen revealed active glucose-6-phosphatase activity. In group I rats, the survival was longer and that group had more long-term survivors than those of group II controls. In group I, there was significant increase in the ratio of weight of remnant liver lobes to body weight. At 24 hours after hepatectomy, group I rats had improved biochemical parameters compared to those of group II rats. CONCLUSIONS: In rats with acute liver failure, intrasplenic hepatocyte transplantation acts as a bridge to support experimental rats from acute liver failure to liver regeneration, prolong survival in rats with acute liver failure and improve biochemical parameters. PMID- 10791193 TI - Image-navigated endoscopic surgery: introduction of the face-mounted display system with picture-in-picture capabilities. AB - BACKGROUND/AIMS: The face-mounted display system with picture-in-picture capabilities was evaluated for clinical usefulness in endoscopic surgery. METHODOLOGY: The device mounts two crystal liquid color panels, being shaped like a pair of glasses. It can provide us with an ultra-large virtual screen of the endoscopic view. Furthermore, a picture-in-picture capability can materialize a multi-image environment. Twenty-five patients were enrolled in this study. RESULTS: The view obtained by the surgeon consisted of the FMD screen of the upper two-thirds and the direct view of the lower one-third. The surgeon could concentrate more on the procedure, because he could face the target organ regardless of the display position. No complications attributable to the device were encountered. With a multi-image environment, endoscopic procedure was executed without taking one's eyes off the operative field. CONCLUSIONS: The face mounted display system will conduct us to another sphere of endoscopic surgery, i.e., an image-navigated endoscopic surgery. PMID- 10791195 TI - Multiple small lesions of hepatocellular carcinoma controlled by percutaneous and laparoscopic ethanol injection--a case report. AB - A 61-year-old man was admitted to our clinic for a liver examination. Ultrasonography revealed multiple echo-rich lesions in both lobes. A laparoscopy showed a liver with an irregular surface, and a 3-mm-sized dark reddish lesion on the inferior surface of the right lobe. alpha-Fetoprotein and plasma protein induced by vitamin K absence or antagonist were normal. A liver biopsy specimen obtained from the small lesion by laparoscopy-guide showed a well-differentiated hepatocellular carcinoma with bile formation. Biopsy specimens obtained later from the 2 echo-rich lesions by ultrasonographic-guide were histologically similar to the lesion laparoscopically observed. Laparoscopic ethanol injection and percutaneous ethanol injection were performed as therapeutic procedures. Recurrence of hepatocellular carcinoma at the treated sites was not observed during the 6-year observation period. Thus, laparoscopy might play an important role in the early detection and treatment of hepatocellular carcinomas on the surface of the liver. PMID- 10791194 TI - Stop-flow in mediastinum and thorax for resistant lymphoma. AB - BACKGROUND/AIMS: Management of patients with heavily pretreated malignant lymphoma failing frontline treatment and salvage high-dose chemotherapy and autologous peripheral stem cell rescue, is problematic. A pilot study was conducted to evaluate isolated thoracic perfusion of drugs by means of stopflow technique. METHODOLOGY: Six patients were enrolled in the study; diagnoses included 4 advanced Hodgkin's disease, 1 primary mediastinal B-cell lymphoma, and 1 anaplastic large cell lymphoma. Patients were aged 18-37 years; 4 presented with bulky mediastinum. They had never achieved a complete response since all had progressed from front-line treatment, and 3 had even failed salvage high-dose chemotherapy with autologous peripheral stem cell rescue. Cisplatin (100 mg/m2) and melphalan (35 mg/m2) were used. Carmustine (100 mg/m2) were added to these 2 drugs and cytarabine (2000 mg/m2) in patients not previously treated by carmustine, etoposide, cytarabine, and melphalan. Epidoxorubicin (70 mg/m2) was added in patients who previously received a suboptimal dosage of antracycline. Drugs were delivered monthly via aortic perfusion performed by means of Aigner's stop-flow technique. RESULTS: Overall 13 cycles of perfusional chemotherapy were administered with a median number of 2 cycles. During the procedures there were no technical, hemodynamic, or vascular complications, and no deaths occurred during surgery. After 1 month, 6 (100%) objective responses after isolated thoracic perfusion were recorded, 3 (50%) of which were complete. Tolerance to therapy was excellent. Hematological toxicity was mild and transfusional support was needed only in one course. At the last follow-up, 2 patients are alive (1 complete response and 1 very good partial response, maintained). CONCLUSIONS: This new therapeutical approach seems very active in recurrent/refractory malignant lymphoma and may play an important role in this setting. PMID- 10791196 TI - A comparative study of immunochemical fecal tests for detection of colorectal adenomatous polyps. AB - BACKGROUND/AIMS: This study was conducted to assess the diagnostic validity of new different immunochemical fecal occult blood tests for colorectal adenomatous polyps, including reversed passive hemagglutination test (Immudia-HemSp), combination monoclonal antibody guaiac test (Monohaem), latex agglutination inhibition test (Iatro Hemcheck), and latex agglutination tests (LA Hemochaser, OC-Hemodia). METHODOLOGY: Two hundred and fifty patients with colorectal adenomatous polyps 1.0 cm or larger in diameter and the same number of healthy controls served as subjects for the study. Each subject received a set of 5 immunochemical tests within 3 consecutive days, and sensitivities and specificities of these tests were evaluated. RESULTS: Mean sensitivity and specificity in a set of 5 immunochemical tests were 47.6% and 95.8%, respectively. Sensitivities and specificities of 5 different immunochemical tests were calculated as 47.6% and 96.8% for Immudia-Hem Sp, 46.8% and 95.2% for Monohem, 46.4% and 95.2% for Iatro Hemchek, 48.0% and 96.4% for LA Hemochaser, and 49.2% and 95.6% for OC-Hemodia, respectively, showing no significant difference in sensitivity and specificity among the 5 tests. CONCLUSIONS: These findings suggest that there is no difference in the degree of accuracy for colorectal adenomatous polyps among several types of immunochemical fecal occult blood test. PMID- 10791197 TI - Effects of elemental diet (ED) on surgical treatment in Crohn's disease. AB - BACKGROUND/AIMS: Patients with Crohn's disease are at risk of developing short bowel syndrome if they are treated with repeated operations. We examined the effect of preoperative nutritional therapy with elemental diet on the interval until and the outcome after surgical treatment in patients with Crohn's disease. METHODOLOGY: We reviewed the records for 73 patients with Crohn's disease who underwent intestinal resection in our department between January 1, 1974 and December 31, 1996. Thirty-two of them had not received elemental diet (No ED group) and 41 had received elemental diet therapy (ED group) before the first resection. RESULTS: The median interval between the onset and first resection was 19.3 months for the NoED group and 67.5 months for the ED group (P = 0.0005). All patients received ED therapy after the first resection. The incidence of second resection was significantly different between the ED and the NoED group after the first operation. (P = 0.045). CONCLUSIONS: Elemental diet therapy is effective in prolonging the interval between the onset and first resection and in reducing the incidence of second resection. The initial introduction of elemental diet therapy is useful in the management of the clinical course of Crohn's disease minimizing surgical intervention. PMID- 10791198 TI - First clinical data of a natural immunomodulator in colorectal cancer. AB - BACKGROUND/AIMS: MSC (trade-name AVEMAR) is a per os applicable complex of multiple, biologically active molecules obtained from fermented wheat-germ extract. Preclinical studies suggest potent anti-metastatic activity and it has a favorable toxicity profile. It has been aimed in a pilot-scale, phase II clinical study to document whether or not MSC as a support to surgery or plus chemotherapy adds any therapeutic benefit compared to the same combination without MSC in colorectal cancer. METHODOLOGY: From 1998 to June 1999, 18 control patients and 12 consecutive colorectal cancer patients respectively, were enrolled into this study. All patients underwent curative surgery. The control group (18 patients) received no other therapy or adjuvant chemotherapy alone. The MSC group (12 patients) received MSC alone or plus adjuvant chemotherapy. Until now, the median follow-up has been 9 months. RESULTS: Interim data of the study document that in the MSC group no new metastases, neither hepatic nor other, have occurred, so far. On the contrary, several new metastases have developed in the control group. CONCLUSIONS: Orally administered MSC is a potent candidate to be regarded as a supportive therapy to surgery or plus chemotherapy for colorectal cancer patients. PMID- 10791199 TI - Cost-effectiveness analysis of immunochemical occult blood screening for colorectal cancer among three fecal sampling methods. AB - BACKGROUND/AIMS: To evaluate the optimal sampling times of stool in immunochemical occult blood screening from the viewpoint of the cost effectiveness. METHODOLOGY: A colorectal cancer screening was conducted in 5 municipalities (Matsumoto, Hata, Yamagata, Asahi, Sakai), Nagano prefecture, Japan in 1996. Each participant received a fecal occult blood test with 3 consecutive days. For the economic assessment of testing methods, the results of the 1st day, those of the 1st and 2nd days and those of the 3 consecutive days were used for a single-day method, a 2-day method, and a 3-day method. The average costs per detection of one cancer patient and diagnostic validity were evaluated among 3 months. RESULTS: The average costs for one cancer case detected were calculated to be $5924 for a single-day method, $6014 for a 2-day method, and $7123 for a 3-day method, respectively. The sensitivity and specificity were calculated to be 58% and 96% for a single-day method, 89% and 95% for a 2-day method, and 100% and 94% for a 3-day method, respectively, indicating a significant difference in the sensitivity between a single-day method and a 2-day as well as a 3-day method (P < 0.05), and in the specificity among 3 testing methods (P < 0.001). CONCLUSIONS: The present analysis suggests that a 2-day collection method is recommended in the immunochemical occult blood screening from the viewpoint of the cost-effectiveness as well as the diagnostic accuracy. PMID- 10791200 TI - Peptidergic nerves in the colon of patients with ulcerative colitis. AB - BACKGROUND/AIMS: The cause of impaired motility, such as diarrhea and toxic megacolon, in patients with ulcerative colitis is unknown. Neuropeptides have recently been shown to be a neurotransmitter in the non-adrenergic non cholinergic inhibitory and excitatory nerves in the human gut. To clarify the physiological significance of vasoactive intestinal polypeptide, substance P and neurotensin in the colon of patients with ulcerative colitis, we investigated the enteric nerve responses on lesional and normal bowel segments derived from patients with ulcerative colitis and patients who underwent colon resection for colonic cancers. METHODOLOGY: Twenty-four specimens were obtained from the lesional colon of 6 patients with ulcerative colitis (4 male, 2 female; ages 14 51 years, mean: 40.3 years). The patients with ulcerative colitis had chronic disease (4 with moderate disease, 2 with severe disease). Seventy-two specimens were obtained from the normal colon of 10 patients with colonic cancer (8 men and 2 women; ages 40-56 years, mean: 51.2 years). A mechanographic technique was used to evaluate in vitro muscle responses to these peptides of adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers. RESULTS: (1) Peptidergic nerves such as vasoactive intestinal polypeptide, substance P, and neurotensin nerves were found to act on both normal colon and ulcerative colitis colon; (2) the colon with ulcerative colitis was more strongly innervated by vasoactive intestinal polypeptide nerves than the normal colon; (3) Substance P and neurotensin nerves act more weakly in the UC colon that the normal colon. CONCLUSIONS: These findings suggest that peptidergic nerves play an important role in the impaired motility observed in patients with UC. PMID- 10791201 TI - Detection of colorectal carcinoma by anti-CEA monoclonal antibody (IOR-CEA1) labeled with 99mTc scintigraphy. AB - BACKGROUND/AIMS: This study shows that a new monoclonal antibody (IOR-CEA1) labeled with technetium-99m has high diagnostic efficacy for colorectal adenocarcinoma. This immunoscintigraphy is helpful in clinical management especially for patients whose serum carcinoembryonic antigen and computed tomography are questionable for recurrent diseases. The study aims to evaluate the efficiency of a new monoclonal antibody (IOR-CEA1) labeled with technetium 99m in the detection of colorectal carcinoma. METHODOLOGY: Forty colorectal carcinoma patients were examined. They were divided into 2 groups: Group I (9 patients) with untreated primary tumor; and Group II (31 patients) who were suspected of recurrent or residual diseases from 1) equivocal computed tomography or magnetic resonance imaging, or 2) rising serum carcinoembryonic antigen but normal imaging or clinical findings. One milligram of the antibody labeled with 25mCi of technetium-99m was slowly infused intravenously and images were obtained by nuclear medicine techniques. Sensitivity, specificity, accuracy, positive and negative predictive values were determined. RESULTS: 99mTc-IOR-CEA1 had 86% sensitivity, 71% specificity, 83% accuracy, 94% positive predictive value and 50% negative predictive value for the detection of colorectal cancer in 42 studies (2 patients had repeated studies). Serum carcinoembryonic antigen had only 33% sensitivity for detection of the primary cancer and 58% sensitivity in detection of recurrent diseases. Carcinoembryonic antigen had 100% positive predictive value but only 31.3% negative predictive value for diagnosis of the recurrence of tumor. Fifty-two percent of the antibody scans provided more information than computed tomography scans with clinical impact on further management in group II patients. CONCLUSIONS: The 99mTc-IOR-CEA1 scintigraphy is a promising investigative method which is safe and has high accuracy in the detection of recurrent colorectal carcinoma, especially in the patients whose serum carcinoembryonic antigen and computed tomography findings are equivocal for recurrent diseases. PMID- 10791202 TI - Characteristics and survival rate of elderly patients with colorectal cancer detected by immunochemical occult blood screening. AB - BACKGROUND/AIMS: A comparative study was carried out to clarify the clinicopathological features and survival rate of elderly patients with colorectal cancer diagnosed by immunochemical fecal occult blood screening. METHODOLOGY: The study subjects included 208 patients with colorectal cancer, who were divided into 2 groups based on their ages: 52 patients aged 75 years and over (older group) and 156 patients aged 50-74 years (younger group). The 2 groups were compared in terms of their individual factors, including site, size, Dukes' classification, histo-pathological type of their cancer lesions and 5-year survival rate. RESULTS: There was a significant difference in prevalence of tumor sites between the older and younger groups (P < 0.05). Particularly, the frequency of right colon cancers was higher in the older group (34.6%) than in the younger group (20.5%), but there were no differences found in any other factors such as size, Dukes' classification, and histopathological type. Survival rate was almost the same between the 2 groups. CONCLUSIONS: These results suggest that elderly patients are susceptible to right colon cancer but there is no difference in the clinicopathological characteristics and 5-year survival rate between older and younger patients with colorectal cancer detected by immunochemical fecal occult blood test. The older as well as younger people should be encouraged to attend the screening program for colorectal cancer by immunochemical fecal occult blood test. PMID- 10791203 TI - Immunohistochemical status of the p53 protein and Ki-67 antigen using biopsied specimens can predict a sensitivity to neoadjuvant therapy in patients with esophageal cancer. AB - BACKGROUND/AIMS: The prediction of an effective adjuvant therapy is essential in order to increase the number of esophageal cancer cases demonstrating an excellent response. Correlation of the immunohistochemical expression of p53 protein and Ki-67 antigen were studied using biopsied specimens of advanced esophageal cancer. The combined staining status of both can help predict the efficacy of neoadjuvant therapy in patients with esophageal cancer. METHODOLOGY: The overexpression of p53 protein and the Ki-67 labeling percentage were immunohistochemically investigated in 95 biopsied specimens of advanced esophageal carcinoma resected between 1988 and 1996. All patients preoperatively received 1 course of either chemoradiotherapy or hyperthermo-chemoradiotherapy. RESULTS: Thirty-nine specimens were positive for p53 protein staining (41.1%), and the treatment modalities were histopathologically effective in 71.8% of these 39 patients (28/39), while the efficiency rate was 58.9% (33/56) in patients with p53(-). On the other hand, the efficiency rate of patients with a high Ki-67 labeling percentage (> or = 30%) was 73.9%, which was significantly higher than that of those with a low Ki-67 labeling percentage (< 30%) (38.5%, P = 0.0013). The efficacy rate of the patients with both a high Ki-67 labeling percentage and p53(+) was 80%, while that of the patients with a low Ki-67 labeling percentage and p53(-) was only 35.3% (P = 0.0098). The combination of a high Ki-67 labeling percentage and p53(-) particularly showed a high sensitivity to hyperthermo chemoradiotherapy since the efficiency rate of these patients with hyperthermo chemoradiotherapy was 81.5%, while the rate for those with chemoradiotherapy was 41.7% (P = 0.0129). CONCLUSIONS: An immunohistochemical analysis of p53 and the Ki-67 labeling percentage using biopsied specimens was thus found to effectively predict the efficacy of neoadjuvant therapies in patients with esophageal cancer. PMID- 10791204 TI - Results of sclerotherapy for bleeding esophageal varices in patients with schistosomal liver disease. A retrospective study. AB - BACKGROUND/AIMS: The aims of the study were to evaluate results of injection sclerotherapy in patients with liver schistosomiasis inducing bleeding esophageal varices and to review ultrasonographic features of the liver disease as well as endoscopic characteristics of the esophageal disease in order to assess any interrelationship between them. METHODOLOGY: A total of 34 patients with active or recent history of hematemesis and Schistosoma mansoni infection had emergency or elective endoscopic sclerotherapy. Each underwent ultrasound examination to assess hepatosplenic involvement and staging, and were followed-up with upper digestive endoscopy every 4 months. RESULTS: Obliteration or reduction of the varices in small columns was achieved in 82.3% of cases. During the follow-up period (mean: 10.4 +/- 2.1 months; range: 4-16 months) rebleeding was noted in 2 patients and 2 patients died due to variceal hemorrhage. The relationship between the ultrasonographic periportal fibrosis grade and the endoscopic variceal grade or varices localization was very strong (P < 0.001). A significant difference between grade 1 vs. 3 and 1 vs. 2 of periportal fibrosis and the presence of red signs was also found (P < 0.008). CONCLUSIONS: In view of the results obtained in terms of success rate in obliterating varices, rebleeding and mortality rates, a longitudinal study could be justified to assess the usefulness of prophylactic sclerotherapy for the prevention of the first variceal hemorrhage and in the attempt to prolong survival in patients with Schistosoma-induced esophageal varices. PMID- 10791205 TI - Superficial undifferentiated small cell carcinoma of the esophagus showing an interesting growing pattern in histology. AB - We report a case of superficial undifferentiated small cell carcinoma of the esophagus. The histology of the tumor was interesting; there was squamous cell differentiation within the epithelial layer and undifferentiated small cells growing within the submucosal layer. The tumor had a negative Grimelius reaction, suggesting no differentiation into a hormone-producing carcinoma known as an apudoma (Amine Precursor Uptake and Decarboxylation (APUD)). The serum levels of ACTH and calcitonin were within normal limits. As the patient was elderly and had a history of pleural tuberculosis causing poor pulmonary function, and owing to the fulminant nature of this carcinoma, he underwent blunt dissection of the esophagus by posterior mediastinal gastric pull-up. The patient recovered quickly and was able to be discharged with a good quality of life until the tumor recurred as a liver metastasis 6 months later. PMID- 10791206 TI - Increased risk of infection with Helicobacter pylori in spouses of infected subjects: observations in a general population sample from the UK. AB - BACKGROUND/AIMS: To investigate risk factors for infection with Helicobacter pylori in a general population sample who accepted an invitation to be screened. In particular, to determine whether being the spouse of an infected subject is an independent risk factor for infection after adjustment for other risks. METHODOLOGY: A serological screening test for Helicobacter pylori was offered to 4015 randomly selected subjects registered at a single general practice in Market Harborough, UK. Those invited were also asked to complete a questionnaire if attending for screening; this included information relating to possible risk factors for infection. Results were analyzed in the overall sample of attenders and a subset of married couples, using univariate and multivariate analyses. RESULTS: Complete results were obtained for 1431 (36%) subjects. Age, childhood social class and household crowding in childhood were independent risk factors after adjustment for other variables. Positive status of spouse was an independent risk factor in a subset of 389 married couples (P = 0.005, Odds Ratio: 2.65, 95% Confidence Interval: 1.34-5.25). CONCLUSIONS: Our study confirmed the correlation between childhood living conditions and infection with Helicobacter pylori. In the study population, having a Helicobacter pylori positive spouse was associated with a significant increased risk of being infected. PMID- 10791207 TI - Changing clinicopathological profile of intussusception in Nigeria--a 20-year review. AB - BACKGROUND/AIMS: Intussuception was a common cause of intestinal obstruction at the University College Hospital, Ibadan. A peculiar type characterized by being ceco-colic, and found most commonly in adults, was the commonest variety found, but over the last few years this type of intussusception appears to have reduced in incidence. The aim of the study is to define the relative incidence of intussusception and the contribution of the various types of the overall incidence as seen at the University College Hospital, Ibadan, Nigeria. METHODOLOGY: Review of case notes and the surgical pathology records of all cases of Intussusception seen at the University College Hospital, Ibadan between 1975 and 1994 was done. RESULTS: There was a 48.1% decline in the absolute number of cases seen during the period of the study, and this decline was more among adult cases than infantile intussusception. The mean age of presentation of infantile intussusception was 8 months, while it was 42 years for adults. The classical triad of vomiting, pain and bloody stool was seen in only 15% of cases. Morbidity and mortality rates were high at 18 and 8.5% respectively and this was related to delayed presentation. CONCLUSIONS: The incidence of intussusception has fallen in the community studied and this decline has affected the adult age group and the ceco-colic type of intussusception more. Late presentation is a feature of most cases and is related to the high mortality and morbidity rates seen. PMID- 10791208 TI - Retroperitoneal malignant fibrous histiocytoma surgically removed six times using a harmonic scalpel. AB - This paper describes a patient with retroperitoneal malignant fibrous histiocytoma who underwent surgical removal of the tumor 6 times. It is difficult to remove locally recurrent retroperitoneal malignant fibrous histiocytoma many times by repeat surgery, since the tumor tends to adhere tightly to the surrounding scar tissue. In the 5th and 6th operations we conducted on the patient, a harmonic scalpel was useful for separating the tumor from the scar tissue at the previous surgical site. PMID- 10791210 TI - Better post-resectional survival in female cirrhotic patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Hepatocellular carcinoma is notably more prevalent in male. The purpose of this study was to assess the surgical results in male and female cirrhotic patients. METHODOLOGY: The surgical outcomes of 129 hepatocellular carcinoma patients with cirrhosis, including 109 males and 20 females, who had undergone hepatic resection were studied. The clinical, histologic features, DNA ploidy and proliferative phase fraction of tumor and cirrhotic liver were compared between male and female patients. RESULTS: Female patients had significantly lower incidences of history of smoking (5.6% vs. 52.9%, P < 0.001), alcohol intake (5.6% vs. 42.3%, P = 0.003) and hepatitis B surface antigen positivity (47.1% vs. 73.5%, P = 0.028) than male. Cell-cycle analysis of tumor part revealed female had a significant lower G2M phase fraction (3.4%) than male (5.7%) (P = 0.027). The 1-, 3-, and 5-year disease-free survival rates in male and female patients were 65.5% and 88.2%, 36% and 64.4%, and 29.7% and 64.4%, respectively. Female patients had a significantly better disease-free survival than male (P = 0.034, log-rank test). CONCLUSIONS: Female hepatocellular carcinoma with cirrhosis had lower incidences of hepatitis B surface antigenemia, alcohol abuse and lower DNA postsynthetic phase fraction in tumor tissue than male. Consequently, female hepatocellular carcinoma with cirrhosis had better survival than male. PMID- 10791209 TI - Breath and blood ammonia in liver cirrhosis. AB - BACKGROUND/AIMS: Hyperammonemia causes dysfunction of multiple organs in patients with cirrhosis, including hepatic encephalopathy. Blood ammonia concentrations are monitored with respect to disease progression and efficacy of treatment. Fetor hepaticus, the characteristic breath odor in hepatic encephalopathy has called little quantitative attention to breath ammonia. We studied the dynamics of ammonia metabolism in cirrhosis in terms of the relationship between breath and blood ammonia. METHODOLOGY: Breath and blood ammonia levels were measured simultaneously in 20 cirrhotic patients and in 10 healthy volunteers. Breath ammonia was measured using ammonia electrodes in collected expired air. Helicobacter pylori serum antibody titers were also measured, since the organism produces ammonia. RESULTS: Blood ammonia correlated positively with breath ammonia in patients with cirrhosis. Breath ammonia levels were significantly higher in cirrhotic patients (0.745 ppm) than in controls (0.278 ppm), and higher in cirrhotic patients with hyperammonemia (0.997 ppm) than in those without (0.558 ppm). Breath and blood ammonia decreased together with treatment of hyperammonemia. H. pylori seropositivity was 20% in controls, 27.3% in cirrhotic patients with normal blood ammonia, and 66.7% in those with hyperammonemia. CONCLUSIONS: Breath ammonia measurement may be useful in diagnosis, treatment assessment, and follow-up in hepatic encephalopathy. PMID- 10791211 TI - Ultrastructural study of the remnant liver after extensive hepatectomy in dogs; especially morphological alterations of sinusoidal endothelial cells. AB - BACKGROUND/AIMS: To investigate the role of sinusoidal endothelial cells in the mechanism of the remnant liver dysfunction after extensive hepatectomy, morphological alteration of sinusoidal endothelial cells and cytoprotective effects of prostaglandin I2 analog were studied by ultrastructural analysis. METHODOLOGY: Mongrel dogs (n = 24) were divided into 3 groups according to the extent of hepatectomy and the use of prostaglandin I2 analog preoperatively. Group 1: 70% hepatectomy (n = 8), Group 2: 84% hepatectomy (n = 8), and Group 3: 84% hepatectomy with administration of prostaglandin I2 analog, preoperatively (n = 8). Morphology of hepatic sinusoidal endothelial cells was examined after hepatectomy in all cases. RESULTS: Group 2: Enlargement of the sinusoidal endothelial fenestrations was transitorily occurred immediately after hepatectomy, but were dramatically decreased in size 24 h after the operation. Most sinusoidal endothelial cells were destroyed and separated from the sinusoidal wall. Fibrin deposits and sludge were found in the Disse's space and sinusoidal lumens, resembling liver failure at 24 h after the operation. Group 1 and 3: sinusoidal endothelial fenestrations remained slightly enlarged at 24 h after the operation, but fibrin deposits or sludge were not observed in sinusoidal lumens. CONCLUSIONS: Our data suggest that extensive hepatectomy changed the morphology of sinusoidal endothelial cells. Administration of prostaglandin I2 analog helped maintain structural integrity of sinusoidal endothelial cells, improved hepatic microcirculation and thus served to reduce remnant liver dysfunction. PMID- 10791212 TI - Correlation between a psychometric test and biochemical indices of hepatic encephalopathy in alcoholics. AB - BACKGROUND/AIMS: In alcohol abusers an alteration of responses to psychometric tests has been reported, even when clinical symptoms of hepatic encephalopathy (HE) are absent. Our research was intended to individualize a simple psychometric test, easy enough to be performed also at the patient's home, able to reveal an impending encephalopathy and, consequently, to facilitate earlier treatment. METHODOLOGY: Twenty-six consecutive male alcoholics were engaged and, after informed consent, the following schedule was applied: administration of a psychometric test, followed by a drawing of blood for the determination of many blood parameters. After 15 days of treatment to detoxicate patients, psychometric tests and blood examinations were repeated. RESULTS: The results confirmed that common blood examinations are not useful to monitor brain damage in chronic alcoholism, that a psychometric test is able to demonstrate a therapeutic improvement and that a positive and significant correlation has been observed between BBCA/AAA ratio and WAIS Score. CONCLUSIONS: These preliminary results suggest that it is possible to suspect dangerous biochemical changes by means of a simple psychometric test. PMID- 10791213 TI - Effects of prostaglandin E1 administration during hepatectomy for cirrhotic hepatocellular carcinoma. AB - BACKGROUND/AIMS: Prostaglandin E1 has been used in hepatectomy based on a few limited clinical studies suggesting that PGE1 improves liver function. The aim of this study was to evaluate the effects of PGE1 administration during hepatectomy for cirrhotic hepatocellular carcinoma. METHODOLOGY: Forty-three patients undergoing hepatectomy for cirrhotic hepatocellular carcinoma were divided into 2 groups: hepatectomy with Prostaglandin E1 treatment (PG group; n = 19) and without Prostaglandin E1 treatment (control group; n = 24). Prostaglandin E1 (0.02-0.07 microgram/kg/min) was administered intravenously from beginning to end of surgery in the PG group. RESULTS: There were no significant differences between groups with respect to age, gender, preoperative liver and renal function, or intraoperative variables such as blood loss, weight of resected liver and total clamping time by the Pringle maneuver. No patient had severe postoperative complications. Initial postoperative maximum concentrations of serum total bilirubin, creatinine, and blood urea nitrogen in the PG group were significantly lower than those in the control group. CONCLUSIONS: Prostaglandin E1 administration during hepatectomy for cirrhotic heptocellular carcinoma resulted in improved renal and hepatic function. PMID- 10791214 TI - Intraportal insulin therapy after partial hepatectomy for hepatoma patients with insulinopenia. AB - BACKGROUND/AIMS: This prospective study was carried out in order to assess the role of intraportal insulin infusion after partial hepatectomy in hepatoma patients with insulinopenia. METHODOLOGY: Of the 38 patients who underwent hepatic resection for hepatocellular carcinoma from August 1994 to August 1995, 21 patients took an oral glucose tolerance test with insulin measurement preoperatively. Five patients were treated postoperatively with intraportal insulin infusion, as follows: A 16-gauge catheter was inserted into a reopened umbilical vein and fixed in place by a few absorbable stitches. Regular insulin was administered just after the operation at the rate of 2 units/hour for 2-3 weeks. Blood glucose levels were maintained between 150 and 200 mg/dL. Arterial ketone body ratio was used in monitoring the function of the liver during the perioperative periods. RESULTS: Ten patients (47.6%) were found to be insulinopenic. In the insulin therapy group (n = 5), the arterial ketone body ratio of 3 patients recovered within a few hours after the operation, and that of the remaining 2 patients recovered on the 1st postoperative day. But in the control group (n = 5), none of the 5 cases had an arterial ketone body ratio of more than 0.7 on the day of operation. The arterial ketone body ratio returned to normal on the 1st postoperative day in 2 cases, on the 2nd postoperative day in 1 case, and on the 5th day in 1 case. The arterial ketone body ratio recovery time was shorter in the intraportal insulin therapy group than in the control group (P = 0.042). CONCLUSIONS: Intraportal insulin infusion after hepatectomy via the reopened umbilical vein may be a very simple and safe means of promoting recovery of remnant liver function after hepatectomy in hepatoma patients with insulinopenia. PMID- 10791215 TI - The role of vitamin D3 receptor mRNA in the proliferation of hepatocellular carcinoma. AB - BACKGROUND/AIMS: 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] exhibits effects on cell proliferation and differentiation. METHODOLOGY: We examined the anti proliferative effect of 1,25-(OH)2D3 on human hepatocellular carcinoma cell lines (PLC/PRF/5, HCC-T) and the role of vitamin D receptors in modulating this effect. We also investigated the gene expression of vitamin D receptors in tissues from patients with hepatocellular carcinoma. RESULTS: 1,25-(OH)2D3 inhibited proliferation of PLC/PRF/5 cells that express prominent vitamin D receptor mRNA with time and dose dependent manner. On the other hand 1,25-(OH)2D3 had no anti proliferative effect on hepatocellular carcinoma T cells in which gene expression of vitamin D receptor was little. We next examined vitamin D receptor gene expression in surgically obtained 10 hepatocellular carcinoma tissues by reverse transcriptase-polymerase chain reaction analysis. In all 10 cases, the level of vitamin D receptor mRNA in tumorous tissue was expressed on tumorous tissue. CONCLUSIONS: These results indicate that 1,25-(OH)2D3 may clinically exert its anti-proliferative effect through vitamin D receptors. PMID- 10791216 TI - Orthograde whole gut irrigation with mannite versus paromomycine + lactulose as prophylaxis of hepatic encephalopathy in patients with cirrhosis and upper gastrointestinal bleeding: results of a controlled randomized trial. AB - BACKGROUND/AIMS: In patients with liver cirrhosis and upper gastrointestinal bleeding development of hepatic encephalopathy is a major problem. The aim of the present study was to evaluate the efficacy of the mannite lavage in a controlled randomized trial with respect to the Child-Pugh classification. METHODOLOGY: After initial gastroscopy (+/- sclerotherapy) 39 patients with cirrhosis (18 F, 21 M; age: 57.5 +/- 11.9 yr; Child A: 6, Child B: 16, Child C: 17) and upper gastrointestinal-bleeding were classified according to the Child-Pugh-criteria (A,B,C) and randomized in 2 groups (A,B) for each Child-Pugh level. Patients in group A (n = 18) were initially treated with 2000 mL mannite solution (10%) during the first 2 hours using a naso-gastric tube. Treatment was continued using 2000 mL mannite solution (10%) per day until no rectal blood could be observed. Patients in group B (n = 21) were treated with paromomycine ter in die (1 g tid) and lactulose (10 mL tid). There were no statistical differences between both groups concerning age, sex, Child-Pugh-scores, severity or source of bleeding, initial hemoglobin-levels, number of given blood-transfusions or number of patients with sclerotherapy. RESULTS: Patients in group A were treated with a total of 3325 +/- 1897 mL mannite solution. The application was well tolerated. In addition, kinetics of serum creatinine, potassium and sodium levels did not show any significant changes. No significant differences between both groups could be shown with respect to clinical criteria of encephalopathy according to O'Grady and the length of intensive care unit treatment. Moreover, kinetic of ammonia-levels showed a pronounced decrease (P = 0.05) on day 2 versus day 1 in group A (110.0 +/- 24.2 vs. 156.4 +/- 98 mg/dL) as compared to group B (210.0 +/- 52.7 vs. 162.0 +/- 45 mg/dL). In group A, 6 patients (33.3%) died during the study as compared to 3 patients (14.3%) in group B (P > 0.05). The lethality rate was strongly associated with the larger proportion of Child-C-patients in group A. CONCLUSIONS: The data indicate that whole gut irrigation with mannite is equally efficacious as compared to standard treatment for prophylaxis of hepatic encephalopathy after upper gastrointestinal bleeding in liver cirrhosis. In contrast to previously published controlled studies, no impact of the lavage on the mortality rate or duration of intensive care unit treatment could be shown. With respect to the lower costs for the mannite solution as compared to paromomycine and lactulose (ROTE LISTE, Germany), the mannite lavage should be recommended for the prophylaxis of hepatic encephalopathy after upper gastrointestinal bleeding in patients with liver cirrhosis. PMID- 10791217 TI - Postoperative complications of repeat hepatectomy for liver metastasis from colorectal carcinoma. AB - BACKGROUND/AIMS: When a repeat hepatectomy is possible, it is the most effective treatment modality for recurrent colorectal liver metastasis. The aim of this study was to evaluate the surgical risks of repeat hepatectomy for liver metastasis from colorectal carcinoma. METHODOLOGY: Between 1986 and 1996, 60 patients with hepatic metastasis from colorectal carcinoma underwent surgery in the Department of Surgery I, Oita Medical University. Ten of them underwent a repeat hepatectomy. The cases of these 10 patients were studied retrospectively; in particular, postoperative complications and intraoperative blood loss were compared between the initial and second operation. RESULTS: During the second surgery, recurrence was detected adjacent to the hepatic stump in 9 of the 10 patients. During the initial surgery, 6 underwent non-anatomic resections, and 4 had anatomic resection, including 1 extended lobectomy, 1 lobectomy, and 2 segmentectomies. For the second surgery, 3 had anatomic resections, including 2 lobectomies, and 1 segmentectomy, and 7 underwent non-anatomic resections. There were no mortalities during the initial or second operation. There was no morbidity following the initial surgeries and 7 postoperative complications (intraabdominal abscess, 4 cases; biloma, 3 cases) following the second surgeries. Mean blood loss during the second operation (1044 mL) was significantly greater than during the initial operation (561 mL). CONCLUSIONS: The present results show that repeat hepatectomy for recurrent liver metastasis from colorectal carcinoma resulted in significantly greater intraoperative blood loss and postoperative complications than those of the initial surgeries. The blood loss and complications in the second operation, the one for the recurrence, were directly associated with the fact that the recurrence was so close to the hepatic stump. Since the resection line in the second surgery was adjacent to the hepatic hilus, resection of the lesion caused much more injury to the main bile duct and main portal vein than that caused by the. PMID- 10791218 TI - Highly aggressive policy of hepatic resections for neuroendocrine liver metastases. AB - BACKGROUND/AIMS: Neuroendocrine tumors are usually slow growing and carry a prolonged prognosis. The presence of liver metastases significantly impairs long term survival. The clinical experience with 28 patients admitted since 1981 for liver metastases from neuroendocrine tumors was retrospectively reviewed to analyze the clinical and surgical management and to evaluate their outcome. METHODOLOGY: Surgery was indicated in 25 (89.2%) patients. Three had metachronous metastases. A correct diagnosis of these liver metastases was achieved before laparotomy in 15 (68.1%) of the remaining 22. The primary tumor site, unknown in 14/22 patients, was located during surgery only in 8 (57.1%). RESULTS: Due to tumoral spread, surgery was limited to exploration in 3 cases. Liver resections were performed in 19/22 patients (3 for palliation): 11/19 (57.9%) were major hepatectomies and in 8/19 (42.1%) cases they were accomplished by procedures for removing the primary tumor. Overall, curative procedures were carried out in 16/28 (57.1%). Resections were performed in 6 cases without the knowledge of the primary site. There was no operative mortality. Overall recurrence rate was 50.0%. Four-year actuarial survival was 92.6% after resection and 18.5% for patients that did not receive surgery (P < 0.001). CONCLUSIONS: Our experience confirms that the small number of patients makes the management of liver metastases from neuroendocrine tumors difficult to plan. In consideration of the satisfactory results achieved with an aggressive policy of resection, we advise referral of these patients to specialized liver units where major hepatic procedures, even if extended, can be safely performed. PMID- 10791219 TI - Interleukin-2 as a modulator of 5-fluorouracil in hepatic arterial immunochemotherapy for liver metastases. AB - BACKGROUND/AIMS: Hepatic arterial infusion of interleukin-2-based immunochemotherapy has yielded a high response rate (> 75%) in patients with unresectable liver metastases. In order to clarify the mechanisms that underlie the apparent benefit of combination treatment, the role of IL-2 as a modulator of 5-fluorouracil metabolism was investigated. METHODOLOGY: A single dose of 5 fluorouracil (50 mg/kg) with or without IL-2 (3500 Japan Reference Units/kg) was given via the hepatic artery to rats bearing liver metastases. Thirty minutes later samples of liver metastatic foci or surrounding normal liver tissue were removed for the measurement of thymidylate synthase, thymidine kinase and dihydropyrimidine dehydrogenase activity, and 5-fluorouracil content. RESULTS: 5 fluorouracil levels in tumor were significantly higher than in normal liver. Although the addition of IL-2 reduced 5-fluorouracil levels in both tumor and normal liver tissues by the activation of dihydropyrimidine dehydrogenase, the ratio of tumor/normal liver 5-fluorouracil levels was unchanged. Both thymidylate synthase and thymidine kinase activities were significantly inhibited in tumor tissue when the combination of 5-fluorouracil and IL-2 was administered. CONCLUSIONS: IL-2 increases 5-fluorouracil cytotoxicity through the inhibition of thymidylate synthase/thymidine kinase activities in the hepatic arterial infusion. PMID- 10791220 TI - Transjugular intrahepatic portosystemic shunt versus distal splenorenal shunt--a comparative study. AB - BACKGROUND/AIMS: No general consensus exists regarding the proper surgical management of recurrent variceal bleeding due to hepatic cirrhosis. Transjugular intrahepatic portosystemic shunt and distal splenorenal shunt are increasingly being performed in the management of these patients. The present study was undertaken to compare the efficacy, complications and survival rate of these two procedures. METHODOLOGY: Sixty-seven patients with alcoholic liver cirrhosis of Child-Pugh's class A (n = 22) and class B (n = 45) with recurrent variceal bleeding not controlled by conservative means underwent either transjugular intrahepatic portosystemic shunt placement (n = 35) or a distal splenorenal shunt operation (n = 32). These patients were followed for a mean of 887 +/- 189 days. Both groups were compared with respect to the rates of survival, recurrence of gastrointestinal bleeding, encephalopathy, ascitis, shunt blockade and other relevant biochemical parameters. RESULTS: Patients who underwent a distal splenorenal shunt operation had lower rates of recurrence of gastrointestinal bleeding (6.25% vs. 25.71%), encephalopathy (18.75% vs. 42.86%) shunt blockade (6.25% vs. 68.57%) and lower mean fasting blood ammonia levels (56.70 +/- 7.10 mumol/L vs. 61.70 +/- 5.70 mumol/L). However the rate of ascitis was higher amongst these patients (40.63% vs. 11.43%). There was no significant difference in the midterm survival rates between these groups (81.25% vs. 80.00%). Both procedures were effective in controlling functional renal failure, splenomegaly and features of hypersplenism. CONCLUSIONS: Distal splenorenal shunt operation is a better therapeutic option than transjugular intrahepatic portosystemic shunt placement for control of recurrent variceal bleeding due to hepatic cirrhosis. PMID- 10791221 TI - Clinicopathological characteristics of surgically resected minute hepatocellular carcinomas. AB - BACKGROUND/AIMS: The multistep development of overt hepatocellular carcinoma from very well-differentiated early hepatocellular carcinoma, and of early hepatocellular carcinoma from adenomatous hyperplasia has been strongly suggested. The clinicopathologic and immunohistochemical characteristics of solitary minute hepatocellular carcinomas smaller than 1 cm in size have yet to be clarified. METHODOLOGY: Fourteen minute hepatocellular carcinomas were divided into 2 groups consisting of: 1) hepatocellular carcinoma of hepatitis B surface antigen positive patients (B-HCC) (n = 5), and 2) hepatocellular carcinoma of hepatitis C virus antibody positive patients (C-HCC) (n = 9), then they were all analyzed histopathologically and clinicopathologically. Immunohistochemical studies were also performed using the antibodies against p53 protein. RESULTS: Six of the 14 minute hepatocellular carcinoma were demonstrated to be moderately or poorly differentiated tumors. Among the 8 well-differentiated minute hepatocellular carcinomas, 2 tumors already contained less differentiated components. B-HCC tended to be less differentiated than C-HCC (P < 0.05). Adenomatous hyperplasia was detected in only 2 cases of C-HCC. Small cell liver dysplasia was detected significantly more frequently in C-HCC than in B-HCC (P < 0.05). The prognosis of the 14 minute hepatocellular carcinomas varied considerably. Immunohistochemically, some tumor cells were positive for p53 in 3 cases. CONCLUSIONS: Our study suggests that 1) the multistep carcinogenesis through adenomatous hyperplasia may not be so frequent, 2) De novo carcinogenesis from not only well-differentiated hepatocellular carcinoma, but also from less differentiated hepatocellular carcinoma, especially B-HCC, may be present, 3) the carcinogenesis in the B-HCC cases may behave differently from that in C-HCC cases, and 4) minute hepatocellular carcinomas demonstrate varying prognoses after hepatectomy. PMID- 10791222 TI - Thoracoabdominal approaches versus inverted T incision for posterior segmentectomy in hepatocellular carcinoma. AB - BACKGROUND/AIMS: The purpose of the present study was to clarify the indications for a thoracoabdominal right oblique approach and an inverted L incision in posterior segmentectomy of the liver for hepatocellular carcinoma. METHODOLOGY: Forty-six patients with hepatocellular carcinoma who underwent posterior segmentectomy with thoracotomy were divided into 2 groups according to the incision: thoracoabdominal right oblique incision (group A, n = 17) and inverted L incision (group B, n = 29). The perioperative factors of the patients in the 2 groups were compared retrospectively. RESULTS: No significant differences were found in the preoperative factors between the groups. However, a longer operation time (P = 0.01) and more blood loss (P = 0.02) were noted in group B. Although there was no significant difference between the groups in the overall morbidity rate (P = 0.36), a higher rate of pleural effusion was recognized in group A (P = 0.01). CONCLUSIONS: Thoracoabdominal right oblique incision may be suitable for use as an approach for posterior segmentectomy when manipulation of the left lobe is not required. PMID- 10791224 TI - A case of living-related partial liver transplantation using the right gastroepiploic artery for hepatic artery reconstruction. AB - A 13-year-old boy with liver cirrhosis underwent living-related partial liver transplantation with a left lobe from his mother. A standard hepatic artery reconstruction using the recipient right hepatic artery was anticipated. Unfortunately, the recipient hepatic artery was found to be severely arteriosclerotic and was unsuitable for reconstruction. Instead, the right gastroepiploic artery, measuring 2.0 mm in diameter, was mobilized and was anastomosed to the left hepatic artery of the graft in an end-to-end fashion. Arterial blood flow was satisfactory. The patient's postoperative course was uneventful, and he was transferred to a floor bed on the 5th postoperative day. PMID- 10791223 TI - Treatment of solitary small hepatocellular carcinoma: consideration of hepatic functional reserve and mode of recurrence. AB - BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) recurs frequently after initial treatment. The subsequent prognosis varies with the mode of recurrence. Some patients die of hepatic failure even though the HCC is controlled. We consider the clinical stage (CS), using the modified Child-Pugh classification, to be an important factor influencing the prognosis of these patients. METHODOLOGY: To determine the most effective treatment for HCC, we examined 105 patients with solitary small HCC who were followed-up for more than 1 year after initial treatment. All of them were judged to be cured according to imaging or histological studies. The initial treatments were hepatic resection (n = 43), percutaneous ethanol injection therapy (PEIT, n = 33), and percutaneous microwave coagulation therapy (PMCT, n = 29). The modes of recurrence were divided into intrahepatic metastasis (IM) and multicentric occurrence (MO). RESULTS: Prognosis of MO was superior to that of IM in CS I patients, but there was no difference in prognosis between these modes in CS II. The hepatic resection group had more MO recurrences in CS I patients and more IM recurrences in CS II patients. IM developed frequently after PEIT and PMCT, regardless of the CS. Prognosis with hepatic resection was superior to that of the other treatments in CS I patients, but there was no difference in prognosis among the 3 treatment modalities in CS II patients. CONCLUSIONS: These data indicate that hepatic resection is the first choice for treating HCC in CS I patients, and that PEIT or PMCT is preferable for CS II patients. PMID- 10791225 TI - Control of large defect of inferior vena cava during hepatectomy. AB - Torrential bleeding from a large defect in the inferior vena cava is a dangerous complication of hepatectomy. Instead of mobilization, encircling and clamping of the inferior vena cava for control, the author described a simpler technique of controlling the defect using the surgeon's fingers. In two such cases, the bleeding was successfully controlled. PMID- 10791226 TI - Doppler sonography in hepatolithiasis: a case report. AB - In the treatment of hepatolithiasis, it is important to not only remove all stones and eliminate bile stasis in the biliary tract, but also to remove atrophic hepatic tissue, as such tissue may cause recurrent cholangitis as well as latent cholangiocarcinoma. A 75-year-old woman was diagnosed as having hepatolithiasis with stenosis at the branching of the bile ducts in segment II and segment III by sonography, computed tomography, endoscopic retrograde cholangiography, and magnetic resonance cholangiography. Portograms did not reveal portal branches in segment III but did demonstrate the portal branches in segment II. During the operation, Doppler sonography was done. Doppler sonography did not reveal the portal blood flow in segment III, but did demonstrate a "to and-fro" pattern in the portal branches in segment II, indicating atrophic hepatic tissue with disrupted portal blood flow. Lateral segmentectomy (resection of segments II and III and lithotomy with choledo-chotomy were done. The patient is in good health 2 years 6 months after the operation. Doppler sonography is useful in the detection of disrupted portal blood flow and the diagnosis of hepatic atrophy. This is clearly advantageous in the decision-making about whether to perform a liver resection with hepatolithiasis. PMID- 10791227 TI - Carcinoid tumor of the hepatic duct presenting as a Klatskin tumor in an adolescent and review of world literature. AB - This is a case presentation of a 14-year-old boy with the radiological diagnosis of cholangiocarcinoma occluding the hepatic duct bifurcation. His only symptom at presentation was jaundice and further workup confirmed a mass at the porta hepatis. Surgical treatment resulted in a resection of the hepatic bifurcation tumor with a final pathological diagnosis of a carcinoid tumor of the hepatic duct bifurcation. To our knowledge, this is only the second case presented of a resected carcinoid tumor in adolescence. In this communique we present the above case and review of the world literature of biliary neuroendocrine tumors. PMID- 10791228 TI - Multiple macroregenerative nodules in liver cirrhosis due to Budd-Chiari syndrome. Case reports and review of the literature. AB - Clinical, radiological, histological and immunohistochemical methods were used to define the nature of multiple rapidly growing hepatic nodules in 2 young patients with Budd-Chiari syndrome due to myeloproliferative diseases. In one patient, the arterial hyperperfusion of large nodules was demonstrated by dynamic computed tomography and angiography. The explanted livers of these patients showed multiple well-demarcated nodules up to 3 cm in diameter on the background of liver cirrhosis resulting from chronic hepatic congestion. Histologically, these nodules covered a spectrum ranging from adenoma-like lesions to nodules resembling focal nodular hyperplasia. They consisted of essentially normal hepatocytes, and variably contained fibrous septa including neoductules and large, mostly dysmorphic arteries. Sometimes, they were located close to still patent or recanalized veins. These rapidly growing hepatic nodules are best defined as macroregenerative nodules. The knowledge of this entity may help the physician to avoid misinterpretation of such nodules as carcinomas. PMID- 10791229 TI - Primary hepatic carcinoid tumor: a case report. AB - A case of primary hepatic carcinoid tumor is reported. A 57-year-old woman had no endocrine symptoms. Light microscopic findings revealed tumor cells forming trabecular or insular structures and demonstrating diffuse positive staining by the Grimelius method. Mucin stained with Periodic acid-Schiff (PAS) and alcian blue was partially present in the tumor cells. Immunohistochemically, most tumor cells stained positive with chromogranin A and cytokeratin stains. Intensive and careful investigation before and after the operation revealed no other origin of the tumor. We present this rare case, describe the clinical features of hepatic carcinoid, and discuss its diagnosis. PMID- 10791230 TI - Transient chylous ascites following a distal splenorenal shunt. AB - The development of chylous ascites following abdominal surgery is an infrequent yet alarming complication. We present a patient in whom chylous ascites was diagnosed 6 days after a distal splenorenal shunt. Ten days following bed rest, sodium restriction, and a low-fat diet with medium-chain triglyceride supplementation the ascites resolved. PMID- 10791231 TI - Partial portal arterialization in complete en bloc resection of the hepatoduodenal ligament and left lobe of the liver for hepatic hilar cancer. AB - The complete resection of the hepatoduodenal ligament is associated with enormous surgical invasion, which frequently results in postoperative hepatic dysfunction secondary to interruption of the reconstructed artery. We administered partial portal arterialization by anastomosis of the gastroduodenal artery to the portal vein without reconstruction of the hepatic artery in the complete resection of the hepatoduodenal ligament with resection of the left lobe of the liver in a patient with hilar bile duct carcinoma. After division of the proper hepatic artery, the gastroduodenal artery was anastomosed in an end-to-side fashion to the trunk of the portal vein. After division of the portal vein, to prevent ischemia, a single catheter bypass was inserted into a branch of the mesenteric vein and the another side of the catheter was attached to the hepatic end, of the portal vein. The portal vein was reconstructed with the superficial femoral vein graft. The blood supply to the remaining liver was interrupted for only 15 min during which the proximal end of the superficial femoral graft was anastomosed to the hepatic end of the portal vein. Postoperative liver function has been stabilized and his postoperative course is uneventful without portal hypertension. One month postoperatively, angiography through the vessels nourishing the raised jejunum visualized intrahepatic arteries. PMID- 10791232 TI - Strategy for pancreatic endocrine tumors. AB - BACKGROUND/AIMS: The endocrine tumors of the pancreas are rare diseases and there is no established standard therapy for the liver metastasis of pancreatic endocrine tumors. In this study, the therapy for the pancreatic endocrine tumors was evaluated. METHODOLOGY: The endocrine pancreas tumors of 13 patients had been surgically treated. All primary tumors were completely resected. The liver metastasis was recognized in 4 patients. Partial resection of the liver was performed in 2 patients. Lipiodol-transcatheter arterial embolization was performed for synchronous unresectable liver metastases in the other 2 patients. RESULTS: The patients with no liver metastases survived without recurrence (max: 18.8 yr; mean follow-up: 9.2 yr). The patient with resected synchronous solitary liver metastasis died of recurrent multiple liver metastases 5 months after surgery. The other patient with the metachronous liver metastasis completely resected survived 13.9 years. In the 2 patients with unresectable numerous liver metastases, after lipiodol-transcatheter arterial embolization, tumor necrosis rate was more than 90% in both cases and serum gastrin level was normalized. CONCLUSIONS: Complete resection of liver metastasis is favorable, whereas lipiodol-transcatheter arterial embolization is effective for unresectable liver metastases from pancreatic endocrine tumors as palliation. Complete resection of the primary site is recommended even in the cases with unresectable numerous liver metastases. PMID- 10791233 TI - Pancreaticoduodenectomy for metastatic ampullary and pancreatic tumors. AB - BACKGROUND/AIMS: To report the clinical presentation, diagnosis and results of aggressive surgical management in patients with metastatic ampullary and pancreatic tumors. METHODOLOGY: Twelve patients underwent pancreaticoduodenectomy for ampullary or pancreatic metastases from January 1, 1987, to June 30, 1998, in 2 institutions. The primary cancer was renal cell carcinoma (n = 5), melanoma (n = 2), venous leiomyosarcoma (n = 1), carcinoid tumor (n = 1), colon carcinoma (n = 1), breast carcinoma (n = 1) and small-cell lung carcinoma (n = 1). The mean interval between primary treatment and metachronous pancreatic metastasis was 88 months. In 3 cases, pancreatic metastases were synchronous with the primary tumor. The main symptoms were jaundice (n = 8) and upper gastrointestinal tract bleeding (n = 2). The principal investigations were computed tomography scan (n = 9), arteriography (n = 7), duodenoscopy (n = 6) and fine-needle aspiration (n = 4). A correct preoperative diagnosis was made for 8 patients. RESULTS: In all cases, the pancreatic tumor was resected with intention to cure or provide useful palliation, using pancreaticoduodenectomy for isolated tumors (n = 11) or total pancreatectomy for multiple lesions (n = 1). Three out of 12 patents had positive lymph nodes, and the resection margin was free of disease in all cases. There was no postoperative mortality. Survival after pancreaticoduodenectomy averaged 26 months. Overall survival of patients undergoing pancreaticoduodenectomy was 35% at 2 years and 17% at 5 years. One patient is still alive more than 10 years after pancreaticoduodenectomy. CONCLUSIONS: Pancreaticoduodenectomy can be performed safely, representing a suitable option for resection in patients with symptomatic or late isolated pancreatic metastases in the absence of widely metastatic disease. The best indications are solitary metastases from renal cell carcinoma, sarcoma and neuroendocrine tumors. However, there is no evidence of survival benefit after pancreaticoduodenectomy for synchronous tumors or metachronous tumors from melanoma or colon carcinoma. PMID- 10791234 TI - Combined resection of the portal vein for pancreatic cancer: preoperative diagnosis of invasion by portography and prognosis. AB - BACKGROUND/AIMS: Pancreatic cancer often invades the portal vein because of the anatomical position. Pancreatic cancer with portal vein invasion was not considered operable, and thus the resectability rate was low. METHODOLOGY: Between March 1976 and February 1994, 140 of 243 patients underwent resection, a resectability rate of 58%. A total of 81 (58%) of these patients underwent portal vein resection. We assessed 56 patients in whom the depth of invasion had already been determined histopathologically and whose superior mesenteric arterial portograms were readable. The 56 patients were classified into 4 groups: normal (Type I), stricture on one side of the portal vein (Type II), stricture on both sides of the portal vein (Type III), complete obstruction (Type IV). The length of the longitudinal lesions on portograms was also measured. RESULTS: In 93% (27/29 cases) of portographic Type I or II lesions with longitudinal lesions of 2 cm or less, portal vein invasion was limited to the tunica media. No patients with cancer invasion into the lumen survived more than 1 year. CONCLUSIONS: For patients with pancreatic cancer Type I or II, preoperative portography findings and longitudinal lesions of 2 cm or less, portal vein resection is indicated, and long-term survival can be expected. PMID- 10791235 TI - Recognition of ERC-induced pancreatitis in patients with choledocholithiasis by an analysis of laboratory findings. AB - BACKGROUND/AIMS: This study retrospectively analyses the relative value of various laboratory tests in the diagnosis of procedure-related pancreatitis in patients submitted to endoscopic retrograde cholangiography with sphincterotomy due to choledocholithiasis. METHODOLOGY: A detailed laboratory assessment was performed on 238 patients 4, 10, 24, 48 and 72 hours after undergoing endoscopic sphincterotomy. RESULTS: Of the 238 patients analyzed, 16 (6.7%) had a complication, including pancreatitis in 6 (2.5%), hemorrhage in 4 (1.7%), acute cholangitis in 3 (1.3%), and perforation in 3 (1.3%). In patients developing acute pancreatitis a significant rise was noted not only in serum amylase but also in the white blood cell count and aspargine transaminase at 24 hours following endoscopic sphincterotomy. Endoscopic procedure (difficulty in cannulating the bile duct and/or obtaining cholangiogram) was a significant risk factor for complications. CONCLUSIONS: It is possible to stratify patients according to their risk of developing pancreatitis according to serum amylase, white blood cells and aspargine transaminase levels monitored for at least 24 hours following ERC. Patients for whom the procedure was difficult and selective cholangiogram was not obtained are at increased risk of developing procedure related complications. PMID- 10791236 TI - Metastasis from colon carcinoma in the dorsal pancreas of a patient with pancreas divisum: report of a case. AB - The authors report the case of a 45-year-old woman who suffered from epigastric pain and was operated on 3 years before for colon cancer. The clinical investigation revealed that the patient had colon cancer metastasis to the dorsal pancreas of a pancreas divisum. PMID- 10791237 TI - Pancreatic leiomyosarcoma: clinicopathohistological presentation of a rare tumor. AB - A smooth muscle tumor originating from the pancreas is an extremely rare neoplasm characterized by peculiar histopathologic features and progressive clinical course. We report a case of such a pancreatic neoplasm occurring in a 57-year-old female who underwent surgical resection. The tumor was histologically as well as immunocytochemically analyzed. The spindle cell pattern, vimentin and actin reactivity confirmed smooth muscle differentiation. On the basis of the degree of cellular atypias and the mitotic counts, the malignant potential of the tumor was assessed. Sharp demarcation of the growth appearing characteristic for this type of pancreatic neoplasia with no invasion into the surrounding organs enabled complete excision in spite of the large dimensions of the tumor. PMID- 10791238 TI - Evaluation of pancreas and other abdominal organs by colonoscopic ultrasound. AB - We report a case where colonoscopic ultrasound was used to evaluate the pancreas. In this case the usual method of evaluating the body of the pancreas by upper gastrointestinal ultrasound was unsuccessful because of the presence of a large hiatal hernia. The other abdominal organs evaluated by colonoscopic ultrasound included the ileo-cecal valve, kidney, liver, spleen and prostate. To our knowledge this is the first case where ultrasonic colonoscope has been used to evaluate the body of the pancrease. PMID- 10791239 TI - Is extended lymphadenectomy valuable in palliatively gastrectomized patients with gastric cancer and simultaneous peritoneal metastasis? AB - BACKGROUND/AIMS: The optimal extent of lymphadenectomy in patients with gastric cancer and simultaneous peritoneal metastasis upon non-curative resection remains unclear. The aim of the present study was to evaluate the efficacy of extended lymphadenectomy in palliatively gastrectomized patients with gastric cancer and simultaneous peritoneal metastasis. METHODOLOGY: The significance and limit of extended lymphadenectomy according to the extent of peritoneal metastasis was analyzed retrospectively in 110 patients with gastric cancer and simultaneous peritoneal metastasis who had undergone palliative gastrectomy. RESULTS: Of the 47 patients with P1 metastasis, the median survival period of the 23 patients who underwent extended lymphadenectomy and the 24 patients who underwent limited lymphadenectomy was 21.7 months and 17.2 months, respectively. Of the 63 patients with P2 or P3 metastasis, the median survival period of the 16 patients who underwent extended lymphadenectomy and the 47 patients who underwent limited lymphadenectomy was 10.4 months and 12.8 months, respectively. No significant differences in survival time based on extent of lymphadenectomy were observed either in the patients with P2 or P3 metastasis (P = 0.262) or in those with P1 metastasis (P = 0.277). CONCLUSIONS: The results of the present study demonstrate that extended lymphadenectomy in gastric cancer yields no positive impact on survival upon non-curative resection either in patients with gastric cancer and simultaneous metastases to the adjacent peritoneum (P1) or to the distant peritoneum (P2 or P3). PMID- 10791240 TI - Helicobacter pylori of remnant stomach and optimal dose of amoxicillin for eradicating Helicobacter pylori. AB - BACKGROUND/AIMS: The optimal dose of antibiotics for Helicobacter pylori eradication is not known. The aim of this study was to evaluate optimal dose of antibiotics (amoxicillin) for eradication of H. pylori in the remnant stomach. METHODOLOGY: Biopsy specimens were obtained from 77 patients who underwent gastrectomy for gastric cancer. H. pylori was subsequently diagnosed by rapid urease test and culture. Gastritis was assessed by scoring. Patients with positive H. pylori were eligible for the eradication study. Amoxicillin 750 mg per day for 2 weeks and omeprazole 20 mg per day for 8 weeks were administered to them. Endoscopic reexamination and 13C-urea breath test were performed 12 weeks after the initiation of treatment. RESULTS: The positive rate of H. pylori was 38.9% in the remnant stomach. Eradication rate was 50.0%. Mean dose of amoxicillin in effective cases was 14.1 +/- 1.5 mg/kg/day. This was significantly higher than that in non-effective cases (12.5 +/- 1.5 mg/kg/day). Remnant gastritis was significantly improved after complete eradication. CONCLUSIONS: H. pylori was present in 38.9% of patients who underwent gastrectomy for gastric cancer. The optimal dose of amoxicillin was 15.6 mg/kg/day for 14 days with omeprazole-amoxicillin therapy. PMID- 10791241 TI - Postoperative results of left upper abdominal evisceration for advanced gastric cancer. AB - BACKGROUND/AIMS: In spite of the improvement of surgical techniques, the prognosis of patients with advanced gastric cancer still remains poor. With the aim of achieving en bloc resection of primary tumor, peritoneal dissemination on the greater omentum and lymph node metastasis, left upper abdominal evisceration (LUAE) was performed for 75 patients. In this report, we investigated the prognostic difference between the LUAE group and standard gastrectomy (total gastrectomy + pancreatosplenectomy). METHODOLOGY: In the LUAE group, total gastrectomy was performed with the en bloc resection of the transverse colon, pancreas body and tail, spleen and left adrenal gland. In addition, omental bursa, covering retroperitoneum and pancreas body and tail was resected in combination with greater omentum, transverse mesocolon, and lesser omentum. RESULTS: There were 3 (4.1%) postoperative death in the LUAE group, and 2 (1.7%) in the control group. However, there was no statistical difference in the incidence of postoperative complications between these 2 groups. The overall 5 year survival rates of the LUAE and control groups were 33% and 39%, respectively. There was no statistical survival difference between these 2 groups. Survival difference between the LUAE and control group was not found in terms of tumor location, wall invasion, lymph node status, peritoneal dissemination, and macroscopic type. CONCLUSIONS: From these results, LUAE cannot improve the survival of patients with advanced gastric cancer. Consequently, LUAE should be indicated for T4 tumors, which directly invade into the transverse colon. PMID- 10791243 TI - Differential effects of radical D2-lymphadenectomy and splenectomy in surgically treated gastric cancer patients. AB - BACKGROUND/AIMS: Radical surgery with extended lymph node dissection (D2-LA) is the treatment of choice for gastric cancer patients in Japan. In Western countries results after D2-LA are controversially discussed, as increased D2-LA related complications are reported. The aim of this retrospective study was to analyze the influence of D2-LA and/or splenectomy on the clinical course, morbidity and long-term survival of gastric cancer patients. METHODOLOGY: Included in this series are 243 consecutively treated gastric cancer patients with a median age of 63.8 years. RESULTS: Though 56% of the patients presented with stage III and IV tumors, a resection rate of 95.5% with R0-resections in 73.7% was achieved. D2-LA rate was 66.3% with simultaneous splenectomy in 48.7%. Global morbidity was 41.6%. Surgical morbidity, leakage and abscess rates after curative R0 resections were not influenced by D2-LA (22.5/6.5/6.5% vs. 21.9/9.1/3.0% +/- LA) but by splenectomy (31.6/13.2/11.8% vs. 14.7/2.1/1% +/- Sx). Simultaneous splenectomy was associated with 7 of 9 leakages, 7 of 9 abscesses, and 4 of 5 cases with postoperative pancreatitis. Long-term survival (Kaplan-Meier) was 40.6% for all, and 58.1% for R0 resected patients. It was not influenced by splenectomy (56/60.3% +/- Sx). CONCLUSIONS: Radical D2-LA has no impact on the morbidity of gastric cancer patients, but splenectomy is always linked with elevated complications without improving survival rates. Therefore splenectomy as potential part of D2-LA can only be advocated in patients with direct infiltration of pancreas or spleen, in stage IV or whenever metastatic spread is supposed in the nodes along splenic artery or hilum. PMID- 10791242 TI - Local injection of anti-cancer drugs bound to carbon particles for early gastric cancer--a pilot study. AB - BACKGROUND/AIMS: A new dosage formulation consisting of an anti-cancer drug bound to activated carbon particles was developed for a local injection against early gastric cancer so that the dosage formulation yields chemotherapeutic effects selectively to the lymph node metastases as well as to the primary lesion. METHODOLOGY: As a pilot study, the new dosage formulation, total of 50-200 mg of methotrexate only or total of 200 mg of methotrexate plus 8 mg of mitomycin C, was injected into the primary lesions and the adjacent gastric wall of 8 patients with early gastric cancer, guided by a gastrofiberscope before gastrectomy. The surgically resected specimens were examined histologically for the therapeutic effects on the primary lesion and its nodal metastasis. RESULTS: The therapeutic effects were seen in 2 of 4 lymph node metastases (50%) and 5 of 8 of the primary lesions (63%), as confirmed histologically with degeneration and/or necrosis. CONCLUSIONS: Preoperative local injection of the new dosage formulation will be useful to give chemotherapeutic effects on the potential metastases in the regional nodes as well as to the primary lesion. PMID- 10791244 TI - Indications for paraaortic lymph node dissection in gastric cancer patients with paraaortic lymph node involvement. AB - BACKGROUND/AIMS: The surgical results for stage IVb gastric cancer remain very poor. The purpose of the current study is to reveal indications for paraaortic lymph node dissection in stage IVb gastric cancer patients with paraaortic lymph node involvement by analyzing prognostic factors for 3-year survival of stage IVb gastric cancer patients followed by curative B resection. METHODOLOGY: The 3-year survival in clinicopathologic variables were compared by univariate analysis. Using Cox proportional hazards regression model, independent prognostic factors were identified from 11 variables. RESULTS: Overall 5-year survival in stage IVb was 26.5%; mean survival was 19 months. Using univariate analysis, p53 expression significantly influenced 3-year survival. Using Cox proportional hazards regression model, the number of total positive lymph nodes, the number of positive paraaortic lymph nodes, and p53 expression were independent prognostic factors. CONCLUSIONS: In stage IVb, paraaortic lymph node dissection should be indicated in patients with < or = 10 total positive lymph nodes, and < or = 3 positive paraaortic lymph nodes or p53 expression < or = 50%. This indication can be applied according to the preoperative imaging, the staining of p53 by endoscopic biopsy specimens and the intraoperative microscopic evaluation of dissected lymph nodes. PMID- 10791245 TI - Postoperative evaluation of pylorus-preserving gastrectomy for early gastric cancer. AB - BACKGROUND/AIMS: There have been many reports proposing some advantages of pylorus-preserving gastrectomy for gastric ulcer compared to the conventional distal gastrectomy. However, it is not clear whether similar results will be obtained from the patients with early gastric cancer. METHODOLOGY: Of 50 patients with early gastric cancer, 25 underwent pylorus-preserving gastrectomy under strict criteria and the other 25 underwent distal gastrectomy with Billroth I anastomosis by the same surgeon. The subjects were then interviewed and examined periodically to assess symptoms, food intake, body weight and serum nutritional parameters. Endoscopy and a radioisotope gastric emptying test was performed 1 year after the operation. RESULTS: Many of the patients with pylorus-preserving gastrectomy complained of gastric fullness after meals, resulting in poor food intake; a significant between-group difference was found up to 1 year after the operation. A low incidence of reflux gastritis and slow gastric emptying were confirmed in the patients after pylorus-preserving gastrectomy. CONCLUSIONS: Pylorus-preserving gastrectomy has advantages over distal gastrectomy in terms of the avoidance of dumping syndrome and protection against duodeno-gastric reflux. However, more time was necessary for improved gastric fullness or food intake. Pylorus-preserving gastrectomy should be applied in younger patients with early gastric cancer expecting long survival. PMID- 10791246 TI - Drug-induced gastropathy in elderly Taiwanese. AB - BACKGROUND/AIMS: Drug-induced gastropathy was common in developing countries in which drug consumption was heavy. The aim of this study is to evaluate the difference in the distribution of clinical features and endoscopic findings between elderly and non-elderly and to determine the risk factors of drug-induced gastrointestinal mucosal damage in elderly. METHODOLOGY: Four hundred and fifty patients with gastrointestinal mucosal damages were recruited from the outpatient clinic or emergency room. All patients were confirmed by endoscopic examination with Olympus Videoscope QX-200 or GIF-p20. Patient's clinical symptoms, endoscopic findings and risk factors were collected. Data was analyzed by chi 2 test and expressed as Odds ratio. RESULTS: The age distribution of gastropathy was predominant at 60-69 years old, and the case number gradually declined as age increased or decreased, respectively. The clinical presentation of asymptomatic bleeding was significantly higher in the elderly than in the non-elderly, while epigastric pain combined with dyspepsia or bleeding was not different between elderly and non-elderly groups. The endoscopic findings of gastric ulcer and erosions were significantly predominant in the elderly group, while no difference was found for duodenal ulcer between these 2 groups. Non-steroidal anti inflammatory drugs were the popular drug which lead to gastropathy in both elderly and non-elderly groups. Drugs, especially steroids, history of arthritis or peptic ulcer, and alcohol consumption were found to be the risk factors associated with increased risk of gastropathy in the elderly. Stress was also significantly associated with increased risk of gastropathy in the non-elderly. There was no significant difference in smoking habit and use of other drugs between these 2 groups. CONCLUSIONS: The clinical features of symptoms, endoscopic findings and risk factors of gastropathy varied significantly between the elderly and the non-elderly. Drug-induced gastropathy, especially steroid treatment for arthritis, was a significant risk factor in the elderly. Program for assessment and management of these elderly patients under treatment is important. PMID- 10791247 TI - Malignant endocrine carcinoma of the stomach. AB - Small cell carcinoma and small portions of poorly differentiated adenocarcinoma of the stomach sometimes differentiate into secretory cells. We experienced a pure endocrine carcinoma of the stomach. The patient, who had a giant mass in the greater curvature of the body of the stomach and regional lymph node metastasis, underwent total gastrectomy. Immunohistologic staining of the resected specimens revealed that the tumor was an endocrine carcinoma. The tumor cell shape in the present case was polygonal or oval and the cell size was relatively large. The cell nucleus was coarse chromatic and nucleoli were seen, whereas, the nucleus of small cell carcinoma often shows a fine chromatic pattern and nucleoli are not seen. On the basis of these findings, the present case was considered to be a pure endocrine carcinoma which was not classified as a small cell carcinoma. PMID- 10791248 TI - [Evaluation of hemodynamic reserve capacity of a chronic cerebral ischemia model in rat by reflectance spectrophotometric technique]. AB - The hemodynamics of chronic cerebral ischemia has not well been studied compared to that of acute cerebral ischemia. We assessed the capacity of the hemodynamic reserve of chronic cerebral ischemia in a rat carotid-jugular arteriovenous shunt model. Eight to 12 weeks after creation of the arteriovenous shunt, the sequential changes of concentration and oxygen saturation of hemoglobin (Hb) in the both parietal cortices under resting and hypercapnic conditions were spectrophotometrically measured through a thinned skull window. The relative blood flow index (BFI) was calculated from the chronological concentration curve of indocyanine green (ICG) for evaluation of regional cortical blood flow (rCBF). These procedures were repeated after the shunt closure. Hb oxygen contents for saturation, total Hb concentration during hypercapnia, and BFI on the shunt side were lower than those on the contralateral side. They increased, however, after the closure. These findings showed that capacity of hemodynamic reserve impaired due to chronic cerebral ischemia could be improved by an increase of perfusion pressure. PMID- 10791250 TI - [Oculomotor deficits produced by unilateral chemical deactivation of the cerebellar flocculus in alert cats]. AB - We have reported earlier that in alert cats the cerebellar flocculus contains many Purkinje cells that respond to pitch (vertical) rotation and that the majority of them receive excitation from the contralateral posterior canal. To understand the role of the flocculus in oculomotor function, I first examined the simple-spike activity of floccular Purkinje cells responding to sinusoidal pitch rotation and vertical optokinetic stimuli, and then injected the GABA agonist muscimol into the flocculus to examine oculomotor deficits. The great majority of Purkinje cells responded to upward pitch rotation and downward optokinetic stimuli, with their response phases near the resultant eye velocity. Following muscimol injection into the up-pitch areas (1 microgram), the down vestibulo ocular reflex (VOR) induced by up-pitch was depressed at low (not high) stimulus frequencies. Down optokinetic responses were virtually abolished. The cats failed to maintain downward gaze position, and postsaccadic exponential centripetal drift appeared with the mean time constant of 0.7 s. Ocular torsion also appeared on the eye ipsilateral to the injection side in 2 of 3 cats examined. These results indicate the involvement of the flocculus in down and torsional slow eye movements. PMID- 10791249 TI - [The role of beta 2-adrenoceptor gene polymorphisms in asthma]. AB - It has been reported that beta 2-adrenoceptor polymorphisms at nucleotide 46 (Arg16 to Gly) and nucleotide 79 (Gln27 to Glu) enhance and suppress receptor downregulation after prolonged exposure to a beta 2-agonist in vitro, respectively. To elucidate the role of these polymorphisms in bronchial asthma, the author studied the following three points: (1) the prevalence of these polymorphisms in the Japanese population, (2) whether these polymorphisms influence the clinical characteristics of asthma and (3) whether Arg16 to Gly polymorphism causes changes in baseline airway tone, methacholine airway responsiveness, and bronchoprotective and non-pulmonary effects of a beta 2 agonist after 2 weeks of regular, inhaled beta 2-agonist therapy in mild and moderate asthmatic patients. The allele frequencies of Gly16 and Glu27 in the Japanese healthy population were 0.49 and 0.09, respectively; both were significantly lower than those of Caucasians. The percentage of the Gly16/Gly genotype was significantly higher in moderate to severe asthma than in mild asthma (14.0 and 33.3%, respectively, p < 0.05). After regular inhaled salbutamol, a significant decrease of FEV1 (p < 0.05) and a significant increase of airway responsiveness (p < 0.02) were observed in the patients with Gly16/Gly. Although there were no significant changes in the bronchoprotective effect of salbutamol among the three genotypes, the increase in plasma cAMP induced by inhaled salbutamol was significantly attenuated in the patients with Gly16/Gly (p < 0.05). These results suggest a racial difference in the prevalence of beta 2 adrenoceptor polymorphisms that may be important modifiers of clinical characteristics of bronchial asthma. PMID- 10791251 TI - [Molecular and pathological analyses of newly established transgenic rats carrying human endogenous retrovirus gene, ERV3]. AB - Endogeneous retroviruses are known to be integrated in eukaryotic genome as proviral DNA similarly to infectious retroviruses. They are present in many kinds of living things, but their functions, especially those in humans, are unclear. To investigate the function of human endogeneous retroviruses, we chose endogeneous retrovirus type 3 (ERV3) which is a single copy type of human endogeneous retrovirus with mRNA expression in organ tissues in vivo. The full provirus genome of ERV3 was subcloned as a transgene and two lines of transgenic rats carrying ERV3 gene (ERV3 rats) were established. One line of ERV3 rats, ETR5, showed tandem insertion of multiple copies of the transgene and expressed ERV3 mRNA in various organs. High levels of the mRNA expression were detected in both lacrimal and salivary glands. In placentas, where ERV3 is expected to express at high levels in humans, the mRNA expression was evident from 12 days gestation in ETR5 rats. Another line, ETR16, showed a single copy insertion and expressed ERV3 mRNAs only in the lacrimal and salivary glands. By Northern analysis, the expected size (3.5 kb) of ERV3 env transcription as was already shown in human tissues was confirmed in ETR16, but high expression of an additional large transcript (4.0 kb) was detected in ETR5. The result of RT-PCR analysis of the transcript in ETR5 indicated that the tandem insertion in ETR5 genome probably caused mis-promoting and mis-terminal poly(A) splicing in the 3'LTR, resulting in the extension of ERV3 env transcript to 4.0 kb mRNA with an addition of nontranslated LTR sequence. By Western blot using an antiserum against oligopeptides synthesized from ERV3 env sequences, protein product of the transgene was shown to be an 85 kDa band in the lacrimal gland of ETR5. Although no pathological significance was evident in these transgenic rats under conditions without any treatment, ERV3 rats may be a suitable model for analysis of the ERV3 function in vivo. PMID- 10791252 TI - [Chronological analyses of neuropathological and molecular biological changes in affected spinal cord of HTLV-I-infected rat (HAM rat disease)]. AB - The author chronologically analyzed neuropathological aspects of the demyelination process in spinal cords of a rat model of human T-cell leukemia virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) from early asymptomatic to late disease stage for clarifying the pathogenetic roles of HTLV-I in central nervous system. There was no significant difference in histopathological and immunohistochemical findings between the rats within 7 months after the HTLV-I infection and age-matched controls. The first sign of demyelination was the appearance of apoptotic cell death beginning at 7 months after the infection and the apoptotic cell number gradually increased to 12 cells per a whole horizontal section of the spinal cord, in contrast to 5 cells in the control rats. The majority of the apoptotic cells were shown to be oligodendrocytes by immunohistochemical stain with an anti-myelin oligodendrocyte glycoprotein antibody. Increment of activated microglia/macrophages started at 9 months after the infection and they rapidly increased from 15 months to reach 600 cells per a whole horizontal section, in contrast to 300 cells in the control rats. Rapid increase of gemistocytic astrocytes was found from 20 months after the infection (the late disease stage). Molecular analysis of the spinal cords revealed that HTLV-I provirus DNA was evident as early as 4 months after the infection, and massages of HTLV-I pX and tumor necrosis factor (TNF)-alpha began to be expressed at 7 months, just before or at the same time as the appearance of the apoptotic cells. The collective evidence suggests that the apoptotic death of oligodendrocytes, which may be induced either directly by the local expression of HTLV-I or indirectly by upregulated cytotoxic humoral mediators, such as TNF alpha, through the transactive function of p40 Tax, is the major cause of chronic progressive myelopathy in WKAH rats with HTLV-I infection. PMID- 10791253 TI - Using HOS for characterization of periodical and time-varying waveform. AB - We applied an approach based on high order statistics (HOS) to classification of time series. This is different from most of the prior analyses in which signal features have been compared. In our approach, we investigate an input signal deformed non-linearly in time and contaminated with additional different forms of noise, e.g. Gaussian, non-Gaussian and their mixed form. The bispectrum-averaging technique (third order statistics) which preserves information about both amplitude and phase of the Fourier transform is studied for the reconstruction of the predictable waveform. The technique does require neither an initial estimate nor a peak-to-peak synchronization of the realizations. Thus, the normal waveform is considered ideally to be smooth and entirely predictable. The residual is characterized by subtracting the predictable part from the original waveform. Under non-linearity assumption, we propose that the Volterra process tool is suitable for analysis and differentiation between the patterns contained in the time series residual. We present and classify some results of the output of the linear transfer function of the process. The approach is numerically tested with the use of computer-generated data. Its effectiveness in dealing with electrocardiogram (ECG)-related data is also demonstrated. PMID- 10791254 TI - Immune mechanisms of uveitis: insights into disease pathogenesis and treatment. PMID- 10791255 TI - HLA testing in patients with uveitis. PMID- 10791256 TI - Diagnostic testing in patients with ocular inflammation. PMID- 10791257 TI - Treatment strategies in patients with anterior uveitis. PMID- 10791258 TI - Diagnostic and therapeutic approach to patients with retinal vasculitis. PMID- 10791259 TI - Advances in diagnosis and management of herpetic uveitis. AB - Herpetic eye disease is common and is frequently associated with intraocular inflammation or uveitis. Despite recent advances in measuring anti-herpes virus antibodies and viral DNA in ocular fluids, diagnosis remains largely clinical. The two more common syndromes include anterior uveitis, often associated with keratitis, and the acute retinal necrosis (ARN) syndrome. Treatment is complex and requires careful monitoring to provide the appropriate balance of antiviral medication and corticosteroids. Long-term prophylaxis with oral antiviral agents may be required in selected patients to help prevent the vision-compromising complications associated with recurrences. PMID- 10791260 TI - Ocular neovascularization in patients with uveitis. AB - Neovascularization is an infrequent but serious complication of uveitis. The retina and optic disk appear to be affected most often, although new blood vessels may arise from the iris, ciliary body, and choroid as well. Although neovascularization can usually be identified on careful clinical examination, some patients may require fluorescein angiography or UBM. Numerous neovascular growth and inhibitory factors have been identified experimentally. Clinically, however, uveitic neovascularization appears to be determined most directly by the severity of the inflammation and the presence of retinal nonperfusion. Virtually all patients with uveitic neovascularization deserve a trial of local or systemic corticosteroids. Laser photocoagulation can be considered in those patients who fail to respond to corticosteroid therapy, but only when retinal nonperfusion has been demonstrated on fluorescein angiography. Surgical excision of newly formed vessels is reserved for selected patients with CNV and uveitis, but should only be considered when corticosteroids and focal photocoagulation are ineffective or are otherwise contraindicated. PMID- 10791261 TI - Cytomegalovirus retinitis in the era of highly active antiretroviral therapy. PMID- 10791262 TI - Infectious causes of uveitis in the developing world. AB - Infectious causes of uveitis are common in the developing world and include some causes that are rarely encountered in industrialized nations, such as tuberculosis, leptospirosis, leprosy, onchocerciasis, and cystercicosis. Ocular toxoplasmosis occurs in all countries but is more common in Central and South America, the South Pacific, and western Europe. AIDS-related opportunistic infections occur wherever HIV infection is prevalent, including North and South America, western and eastern Europe, the former Soviet Union, sub-Saharan Africa, and South and Southeast Asia. Physicians who care for patients in the developing world should consider these infectious possibilities whenever their patients develop uveitis. PMID- 10791263 TI - Treatment strategies in patients with posterior uveitis. PMID- 10791264 TI - Cataract surgery in patients with uveitis. PMID- 10791266 TI - New and emerging treatments for patients with uveitis. PMID- 10791265 TI - Appraisal and management of ocular hypotony and glaucoma associated with uveitis. PMID- 10791267 TI - [Significance of exercise-induced ST segment depression in patients with myocardial infarction involving the left circumflex artery: evaluation by exercise thallium-201 myocardial single photon emission computed tomography]. AB - The significance of exercise-induced ST segment depression in patients with left circumflex artery involvement was investigated by comparing exercise electrocardiography with exercise thallium-201 single photon emission computed tomography(Tl-SPECT) and the wall motion estimated by left ventriculography. Tl SPECT and exercise electrocardiography were simultaneously performed in 51 patients with left circumflex artery involvement(angina pectoris 30, myocardial infarction 21). In patients with myocardial infarction, exercise-induced ST depression was frequently found in the V2, V3 and V4 leads. In patients with angina pectoris, ST depression was frequently found in the II, III, aVF, V5 and V6 leads. There was no obvious difference in the leads of ST depression in patients with myocardial infarction with ischemia and without ischemia on Tl SPECT images. In patients with myocardial infarction, the lateral wall motion of the infarcted area evaluated by left ventriculography was more significantly impaired in the patients with ST depression than without ST depression(p < 0.01). Exercise-induced ST depression in the precordial leads possibly reflects wall motion abnormality rather than ischemia in the lateral infarcted myocardium. PMID- 10791268 TI - [Clinical significance of pressure measurement in the infarct-related coronary artery in acute myocardial infarction: evaluation of variables predicting recovery of left ventricular function in the convalescent stage]. AB - Early reperfusion and good antegrade flow are essential in restoring better regional left ventricular function in acute myocardial infarction, but they do not always correlate with the extent of recovery. This study evaluated coronary circulation using the new "pressure wire" technique to measure the direct pressure of the coronary circulation including antegrade and collateral flow before and after reperfusion in patients with acute myocardial infarction, and to clarify the influence of these variables on recovery of left ventricular function in the convalescent stage. Fifty six consecutive patients with first acute myocardial infarction underwent percutaneous transluminal coronary angioplasty(PTCA) for totally occluded or severely narrowed infarct-related lesion and evaluation of coronary circulation using pressure wire. Left ventriculography was analyzed at 1 month after the onset in 41 patients. Treatment variables including reperfusion time, reperfusion modality, Thrombolysis in Myocardial Infarction(TIMI) grade after PTCA, and pressure wire variables were compared with parameters of left ventricular function. Reperfusion time was not related to regional wall motion evaluated by the SD chord of left ventriculography in the infarcted zone. Pressure wire measurements showed a correlation between fractional flow reserve measured after PTCA and infarcted regional wall motion(r = 0.558, p < 0.01). Patients with infarct-related lesion in the right coronary artery showed the magnitude of left ventricular regional wall motion was related to fractional collateral flow reserve(maxQc/Qn) during PTCA(r = 0.768, p < 0.05), but no such relationship was observed in patients with infarct-related lesion in the left anterior descending artery. Fractional flow reserve measured after PTCA varied widely in patients with the same TIMI flow grade, so did not vary with it. The pressure wire technique enables assessment of the collateral circulation distal to infarct-related lesion quantitatively before reperfusion in patients with acute myocardial infarction. The fractional flow reserve derived by coronary pressure after reperfusion was significantly related to the recovery of regional wall motion in the infarcted area in the convalescent stage. The fractional flow reserve after reperfusion with PTCA is a better parameter than TIMI flow grade for predicting recovery of regional left ventricular function after myocardial infarction. PMID- 10791269 TI - [Risk factors and effect of reperfusion therapy on left ventricular free wall rupture following acute myocardial infarction]. AB - Reperfusion therapy is one of the most effective treatments for acute myocardial infarction, but the effect on left ventricular free wall rupture remains to be determined. This study tried to clarify the risk factors and effect of reperfusion therapy on the risk of free wall rupture following acute myocardial infarction. 2,671 consecutive patients with acute myocardial infarction admitted to our hospital were examined. Incidence of free wall rupture showed no degenerative change(0 to 5.8%; mean 2.1%). The 1,269 consecutive patients from 1985 to 1995 were examined closely to evaluate risk factors and the effect of reperfusion therapy on the risk of free wall rupture. Fourteen patients who underwent emergent coronary artery bypass surgery were excluded. Free wall rupture was found in 25 patients (2.0%). Multivariate analysis confirmed that high age(> or = 70 years) and first acute myocardial infarction were independent risk factors of free wall rupture (odds ratio 3.62, p = 0.003; odds ratio 7.69, p = 0.046, respectively). The incidence of free wall rupture in the conservative therapy group(n = 799) was 2.1%, successful reperfusion group(n = 373) was 0.5%, and unsuccessful reperfusion group(n = 83) was 7.2% with significant statistical differences(p < 0.01). There was no statistical difference between the direct percutaneous transluminal coronary angioplasty group(n = 84, 3.6%) and the thrombolysis group(n = 372, 1.3%). Successful reperfusion was the only independent factor in the reperfusion therapy group that reduced the incidence of free wall rupture(odds ratio = 0.07, p = 0.001). We conclude that reperfusion of the infarct-related artery and more intensive management of unsuccessful reperfusion is important to prevent free wall rupture following acute myocardial infarction. PMID- 10791270 TI - [Acute myocardial infarction in elderly patients: medical and social problems]. AB - The number of elderly patients with acute myocardial infarction has been increasing. However, the choice of treatment remains controversial. Medical records of 310 consecutive patients with acute myocardial infarction were reviewed. Two retrospective analyses were performed. 1) Patients were divided into the elderly group(70 years or more) and the younger group(under 70 years). In-hospital course and outcome were compared. 2) Pre-hospital performance status and living status were reviewed in the elderly group. Acute phase reperfusion therapy was performed in fewer patients in the elderly group(60.8% vs 71.9%, p < 0.01). The difference was most pronounced in cases of direct coronary angioplasty(28.6% vs 54.7%, p < 0.05). As a result, the rate of reperfusion success(74.8% vs 86.8%, p < 0.01) was lower in the elderly group. Moreover, the rates of in-hospital death(23.6% vs 6.8%, p < 0.005), pulmonary edema(20.3% vs 10.8%, p < 0.05), cardiogenic shock(11.9% vs 6.0%, p < 0.005), pneumonia(17.3% vs 3.0%, p < 0.005), and delirium(29.4% vs 12.0%, p < 0.001) were higher in the elderly group. Five patients in the elderly group and 3 patients in the younger group required rehabilitation because of worsened performance status. Six of them were non-reperfused patients. Elderly patients considered likely to become bed ridden because of pre-existing physical disability at admission accounted for 28.9% of the total. Moreover, many elderly patients had poor support systems (8.4% were living alone, 21.0% were living only with their spouse or a child, 30.1% were widows or widowers). These results show that a lower acute phase reperfusion rate(especially angioplasty) resulted in a poor prognosis and worse performance status in elderly patients. Also 30% of patients were not good candidates for conventional treatment because of delirium, and that self-help in daily life is a fundamental goal for most elderly patients. Rapid and simple acute phase reperfusion, subsequent immediate mobilization, and early discharge are recommended for elderly patients with acute myocardial infarction. PMID- 10791272 TI - [Clinicopathologic study of congenital bicuspid aortic valve in the aged]. AB - Congenital bicuspid aortic valve was found in 25 patients(18 men, 7 women, mean age 78 years) of 7,000 consecutive autopsy cases over 60 years old(0.36%). Nine cases showed aortic stenosis(Group AS), 7 cases showed aortic stenosis and regurgitation(Group ASR) (4 cases with stenosis dominancy, 3 cases with regurgitation dominancy), 9 cases showed no aortic stenosis or regurgitation(Group Non-ASR) and no cases showed pure aortic regurgitation. The groups were compared clinically and pathologically. Cardiac death was frequently found in Groups AS and ASR. The aortic valve was the thickest in Group AS(6.0 +/- 2.1 mm), and thicker in Group ASR (4.6 +/- 1.8 mm) than in Group Non-ASR(2.3 +/- 1.1 mm). Seventeen cases(68%) had right-left cusps of the aortic bicuspid valve and 8 cases(32%) had anterior-posterior cusps. Raphe was seen in 18 cases(72%) and was more frequent in Groups ASR and Non-ASR than in Group AS. Especially, all cases had raphes in Group Non-ASR. Calcification was severe in Group AS and in cases with stenosis dominancy in Group ASR. However, calcification was limited in cases with regurgitation dominancy in Group ASR. Calcification was limited or not present in Group Non-ASR. In this study, the frequency of the bicuspid aortic valve in the aged, the ratio of stenosis to regurgitation, and the distribution of calcification in the valve were established. The bicuspid aortic valve did not necessarily become stenotic, and there were cases having no stenosis or regurgitation. Especially, it was shown that cases without raphe were prone to have marked calcification and to become stenotic. On the other hand, cases with raphe were not prone to become stenotic. PMID- 10791271 TI - [Effect of cholesterol reducing therapy with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor on secondary prevention after myocardial infarction in Japanese patients]. AB - Lowering the blood cholesterol level is a safe method to improve survival for primary and secondary prevention of coronary heart disease. However, there is no evidence for any effectiveness in Japanese. This study was designed to evaluate the effect of cholesterol lowering therapy with 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase inhibitor on cardiac events(death and reinfarction) in Japanese patients after myocardial infarction. A total of 290 patients after myocardial infarction were studied retrospectively. The patients were divided into 2 groups with or without HMG-CoA reductase inhibitor therapy for lowering blood cholesterol levels. The cumulative cardiac events and percentage change of cholesterol levels[total cholesterol and low-density lipoprotein (LDL) cholesterol level] were compared between the 2 groups. HMG-CoA reductase inhibitor therapy lowered plasma cholesterol levels significantly (total cholesterol level--11 +/- 20%, LDL cholesterol level--23 +/- 26%) in patients with hypercholesterolemia, whereas there was no change(total cholesterol level 4.3 +/- 22%, LDL cholesterol level--7.2 +/- 24%) in patients without hypercholesterolemia. HMG-CoA reductase inhibitor therapy reduced cardiac events significantly compared in patients with hypercholesterolemia(p = 0.0008), but there was no benefit in patients without hypercholesterolemia. We suggest that treatment with HMG-CoA reductase inhibitor therapy for lowering cholesterol levels was effective for secondary prevention after myocardial infarction in Japanese patients with hypercholesterolemia. PMID- 10791274 TI - [Usefulness of fluorine-18 fluoro-2-deoxyglucose positron emission tomography for diagnosis and therapy assessment of cardiac sarcoidosis]. PMID- 10791273 TI - [Minimal incision cardiac surgery for atrial septal defects]. AB - Minimal incision cardiac surgery for atrial septal defects was performed in 4 consecutive patients. No special devices or new techniques were used to perform this operation. Clamping of the aorta and snaring of the inferior vena cava could be simply omitted from this procedure. A limited(6 to 7 cm) median skin incision was made, followed by a mini-sternotomy from the second intercostal space to right side of the xiphoid process. Through this limited approach, atrial septal defect closure was performed conventionally with a patch under hypothermic circulatory arrest(less than 10 min). The scar was short(5.5 to 7.3 cm) and cosmetically acceptable in all patients. This mini-incisional procedure is a useful option for atrial septal defect closure that can be completed without sophisticated instruments. PMID- 10791275 TI - [Newly developed heart murmur in a 48-year-old man with heart failure]. PMID- 10791276 TI - Organizational ethics' greatest challenge: factoring in less-reachable patients. PMID- 10791277 TI - Dementia and advance-care planning: perspectives from three countries on ethics and epidemiology. PMID- 10791278 TI - End-of-life decision making: when patients and surrogates disagree. PMID- 10791279 TI - Optimizing ethics services and education in a teaching hospital: rounds versus consultation. PMID- 10791280 TI - Death and remembrance: addressing the costs of learning anatomy through the memorialization of donors. PMID- 10791282 TI - Rational suicide and predictive genetic testing. PMID- 10791281 TI - Attitudes of healthcare professionals toward clinical decisions in palliative care: a cross-cultural comparison. PMID- 10791283 TI - Organ donation and the anguish of failure. PMID- 10791284 TI - The limits of reproductive technology: who decides? PMID- 10791286 TI - Teaching hospitals and Medicare cuts. PMID- 10791285 TI - The need for original ethical analyses for women. PMID- 10791287 TI - Medicare reimbursements for clinical trials. PMID- 10791288 TI - Drug abuse in women. PMID- 10791290 TI - Inhibition of proliferation of human peripheral blood mononuclear cells by calcium antagonists. Role of interleukin-2. AB - To evaluate the role of interleukin-2 (IL-2) in the inhibition of the proliferation of human peripheral blood mononuclear cells (PBMC) by calcium channel blockade, the effect of nifedipine, which blocks the L-type calcium channel on the proliferation, the IL-2 expression and the IL-2 production in human PBMC, was compared with the effect of mibefradil, which blocks both L- and T-type calcium channels with a more selective blockade of T-type channels. The rate of [3H]-thymidine incorporation into control and concanavalin A-induced PBMC in the presence or absence of the calcium channel blockers nifedipine or mibefradil (1, 10 or 50 microM) was assayed in the cells cultured for 3 days. The cellular cytotoxicity and the cell number in growing cultures was also determined in nifedipine- or mibefradil-treated control or stimulated cells. Restoration of the proliferative response in nifedipine- or mibefradil-treated cells was investigated by addition of exogenous IL-2. IL-2 receptor expression in the cells was monitored using antiactivated T-cell antigen (Tac) antibody, and the IL-2 production in the cell supernatants of the cultures was determined by an enzyme amplified sensitive immunoassay. Nifedipine and mibefradil concentration dependently reduced the cell number and [3H]-thymidine incorporation or the do novo DNA synthesis in control and concanavalin A-stimulated human PBMC. The proliferative response of nifedipine- or mibefradil-treated cells was restored by addition of exogenous IL-2. The normal expression of IL-2 receptors was preserved while the IL-2 production was blocked in the presence of nifedipine or mibefradil. PMID- 10791289 TI - Study of the effects of aging on macromolecular synthesis in mouse steroid secreting cells using microscopic radioautography. AB - The effects of aging on DNA, RNA and protein synthesis in adrenal gland cortical cells and testicular Leydig cells of ddY mice at various ages (from prenatal day 19 to postnatal days 1, 3, 7, 14, months 1 and 6 and 1 and 2 years after birth) were examined using light and electron microscopic (EM) radioautography after labeling with [3H]-thymidine, [3H]-uridine and [3H]-leucine. The percentage of the labeled cells in the adrenal glands after [3H]-thymidine injection was greatest in the zona glomerulosa of the cortex and the medulla on embryonic day 19, in the zona fasciculata and zona reticularis of the cortex on postnatal day 1 and gradually decreased with aging. EM radioautography revealed that well developed cell organelles such as smooth surfaced endoplasmic reticulum, Golgi apparatus, mitochondria with tubular cristae and lipid droplets were more frequently observed in the cytoplasm of unlabeled cells as compared to cells labeled with [3H]-thymidine in the three zones of the cortex. The effects of aging on RNA synthesis in adrenal glands after [3H]-uridine injection revealed that all types of cells of the adrenal gland were labeled. In each cell, silver grains were localized over the nuclei and cytoplasm and were more dense in the nuclei than the cytoplasm. Grain counts were highest in the cortex and medulla on fetal day 19 and then gradually decreased with aging from postnatal day 14 to 1 year after birth. In the cortex, the number of silver grains was higher in the zona glomerulosa than in the other zones from fetal day 19 to 1 year and grain counts were higher in the medulla in the embryonic stage as compared to the postnatal stages. However, the number of labeled mitochondria and the mitochondrial labeling index increased with aging. The [3H]-thymidine labeling index in the Leydig cells of the testis was low at embryonic and early postnatal stages, increased slightly at 6 months and reached a peak at 9 months which was maintained at a relatively high level in senescence. The number of the silver grains over the nuclei and cytoplasm of Leydig cells due to [3H]-uridine labeling was observed from embryonic day 19 and increased from month 3 onwards. From adult to senescence, [3H]-uridine incorporation was maintained at high levels in the nuclei and was relatively low in the cytoplasm. The effects of aging on [3H] leucine incorporation in Leydig cells were also examined. The labeling indices between embryonic and early postnatal stages showed no obvious differences although the number of the silver grains in both cytoplasm and nucleus increased from 6 months onwards and was maintained at high levels until senescence. From these results, it was concluded that the effects of aging on DNA, RNA and protein synthesis in steroid secreting cells such as adrenal gland cortical cells and testicular Leydig cells of mice correlated with the hormonal changes observed with aging. PMID- 10791292 TI - Nonadrenergic, noncholinergic responses of the rabbit detrusor smooth muscle and the role of L-arginine/nitric oxide pathway in these responses. AB - In the present study, nonadrenergic, noncholinergic (NANC) responses of rabbit detrusor smooth muscle and the possible involvement of the L-arginine/nitric oxide (L-ARG/NO) pathway was investigated. In the presence of atropine (10(-6) M) and guanethidine (10(-5) M), frequency-response curves were obtained by stimulating tissue with 10-sec trains at increasing frequencies (1-10 Hz) with 3 min intervals between stimulations. Electrical field stimulation (EFS) evoked a biphasic response in rabbit detrusor smooth muscle, consisting of an initial contraction followed by relaxation. ATP desensitization significantly inhibited contractions. L-NAME (10(-5) M) increased the contractions by a maximum of 33 +/- 5% at 1 Hz, 37 +/- 5% at 2 Hz, 18 +/- 4% at 4 Hz, 20 +/- 3% at 8 Hz and 15 +/- 4% at 10 Hz. In detrusor preparations, exposure to L-NAME (NG-nitro-L-arginine methyl ester, 10(-5) M) significantly reduced the maximal relaxation to electrical stimulation to 7 +/- 3% of the control. In the presence of L-ARG (10( 4) M), contractions induced by electrical field stimulation (EFS) at 1, 2, 4, 8 and 10 Hz were reduced by 20 +/- 4, 24 +/- 3, 25 +/- 3, 20 +/- 3 and 30 +/- 5%, respectively. Exposure to L-ARG (10(-4) M) significantly increased the maximal relaxation to electrical stimulation to 52 +/- 5% of the control. Exogenously applied ATP (10(-5)-10(-2) M) to rabbit detrusor muscle resulted in contractions while sodium nitroprusside (10(-7)-10(-3) M) caused concentration-dependent relaxation. These results suggest that nitric oxide (NO) acts as an inhibitory NANC neurotransmitter in the rabbit detrusor smooth muscle. Additionally, NANC contractions mediated by ATP released from NANC nerves, may be masked by the L ARG/NO pathway in this tissue. PMID- 10791291 TI - Effects of genistein and daidzein on enhanced vascular contractile reactivity and Ca2+ sensitivity in cardiomyopathic hamsters. AB - We investigated the contribution of tyrosine kinase activity to the enhanced vasoreactivity in cardiomyopathic (CM) hamsters. The contractile response of the aorta to phenylephrine was greatly enhanced in CM in comparison to control hamsters. Genistein, a tyrosine kinase inhibitor, did not affect the maximum contractile response to phenylephrine in control aortas, although the sensitivity to phenylephrine was significantly diminished. In contrast, genistein markedly inhibited both the sensitivity and the maximum contractile response to phenylephrine in CM aortas. Daidzein, an inactive form of genistein, also inhibited the sensitivity to phenylephrine in both strains although the maximum contractile response was not altered even in the CM aorta. The Ca2+ sensitivity of tension was significantly augmented in alpha toxin-permeabilized smooth muscle from the mesenteric artery of CM hamsters. Furthermore, phenylephrine enhanced myofilament sensitivity to Ca2+ more in CM than control hamsters. The enhancement of myofilament Ca2+ sensitivity by phenylephrine was markedly inhibited by genistein only in CM hamsters. The inhibition by daidzein of the phenylephrine effect on the Ca2+ sensitivity in CM hamsters was less pronounced. These results suggest that an increase in tyrosine kinase activity may lead to enhanced vascular reactivity in CM hamsters possibly due to an increased Ca2+ sensitivity of the contractile apparatus. PMID- 10791293 TI - Dopaminergic involvement in adenosine A1 receptor-mediated antinociception in the tail flick latency model in mice. AB - The interaction between the adenosinergic and dopaminergic systems in nociception was assessed in the tail flick latency (TFL) test in mice. Adenosine exhibited qualitatively different responses depending on the dose: Adenosine 10 and 100 mg/kg i.p. caused antinociception as evidenced by an increase in TFL while the middle dose of 30 mg/kg decreased TFL. On the other hand, the specific adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA) at doses of 0.05, 0.1, 0.25 and 0.5 mg/kg (i.p.) resulted in dose-dependent antinociception. The antinociceptive effect of CPA was reversed by classical albeit nonspecific adenosine receptor antagonist theophylline (5 mg/kg) at a dose which had no effect on TFL per se. A low dose (1 mg/kg i.p.) of the dopaminergic agonist apomorphine caused an early mild hyperalgesic response while the high dose (10 mg/kg i.p.) had no significant effect on TFL. The antinociceptive effect of CPA was attenuated by pretreatment with low dose apomorphine while pretreatment with the high dose caused mild but insignificant potentiation. Theophylline, when administered prior to apomorphine failed to modify the nociceptive response. The results suggest that an interaction between adenosine and dopamine may be involved in nociception. PMID- 10791294 TI - Effects of N6-cyclohexyl adenosine (CHA) on isolation-induced aggression in male mice. AB - Several studies have suggested adenosine receptor involvement in the modulation of aggressive behavior; however, the influence of adenosine A1 agonists on aggression is scarcely known. In this study, we examined the effect of N6 cyclohexyl adenosine (CHA; 0.025-0.4 i.p.), a selective adenosine A1 receptor agonist, on agonistic behavior elicited by isolation in male mice. Individually housed mice were exposed to anosmic standard opponents 60 min after drug administration, and the encounters were videotaped and evaluated using an ethologically based analysis. CHA exhibited an ethopharmacological profile characterized by a selective decrease of offensive behaviors (threat and attack) at the intermediate dose (0.1 mg/kg), without impairment of motor activity. In contrast, the antiaggressive action of the highest doses used (0.2 and 0.4 mg/kg) was accompanied by a marked increase of immobility. It is concluded that the behavioral effects observed in this study could be related to an adenosine modulatory action on other neurotransmitter systems (dopamine/serotonin). PMID- 10791295 TI - Development of apomorphine-induced aggressive behavior: comparison of adult male and female Wistar rats. AB - The development of apomorphine-induced (1.0 mg/kg s.c. once daily) aggressive behavior of adult male and female Wistar rats obtained from the same breeder was studied in two consecutive sets. In male animals, repeated apomorphine treatment induced a gradual development of aggressive behavior as evidenced by the increased intensity of aggressiveness and shortened latency before the first attack toward the opponent. In female rats, only a weak tendency toward aggressiveness was found. In conclusion, the present study demonstrates gender differences in the development of the apomorphine-induced aggressive behavior and indicates that the female rats do not fill the validation criteria for use in this method. PMID- 10791296 TI - Changes in psychic and somatic well-being and cognitive capabilities of peri- and postmenopausal women after the use of a hormone replacement drug containing estradiol valerate and levonorgestrel. AB - A multicenter, prospective, open-label postmarketing surveillance study examined to what extent 2-month oral hormone replacement therapy (estradiol valerate and levonorgestrel; Klimonorm) could produce changes in psychosomatic well-being, self esteem and cognitive capabilities in 78 peri- and postmenopausal women. The women included were 42-58 years of age and had approached the physician due to climacteric symptoms. The following tests were used: Kupperman index, Menopause Rating Scale (MRS II), General Depression Scale (ADS), Zerssen's Symptom List (B L), Frankfurt Self-Concept Scales (FSAL, FSAP, FSEG, FSSW), Digit Symbol Substitution Test (DSST), d2 Test of Attention and Number Square Test. The results showed a clear improvement in subjective psychosomatic well-being and improvements to a lesser extent in the concentration and cognitive capabilities in women in the third treatment cycle. PMID- 10791297 TI - The recovery of cerebrospinal fluid acetylcholinesterase activity in Alzheimer's disease patients after treatment with metrifonate. AB - We investigated the relationship between peripheral and central cholinesterase (ChE) inhibition levels after chronic treatment of Alzheimer's disease (AD) patients with metrifonate (MTF). In a 6-month, double-blind, placebo-controlled trial in AD patients treated with a weekly 2.9 mg/kg MTF dose, we observed 17.15 +/- 23.43, 66.92 +/- 7.30 and 60.80 +/- 12.20% inhibition (n = 6) of cerebrospinal fluid (CSF) and red blood cell (RBC) acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BuChE), respectively. In another study, AD patients were treated with daily MTF to achieve RBC AChE inhibition levels of 85.90%. The CSF AChE inhibition was 67.93 +/- 13.69% (n = 3) at 3-4 h after the last treatment and 6.62 +/- 9.36% (n = 2) at 8 days after dosing. The recovery half time of CSF AChE was 2.21 +/- 1.22 days. These data show that CSF AChE recovers faster than the peripheral plasma and RBC enzymes. Under conditions of chronic weekly dosing with MTF, RBC AChE inhibition does not reflect CSF, and arguably, brain AChE inhibition. Our data do not support continuous central neuronal AChE inhibition as the mechanism for the long-term efficacy of metrifonate for the treatment of AD. PMID- 10791298 TI - Developmental biology protocols. Overview I. PMID- 10791299 TI - Rearing larvae of sea urchins and sea stars for developmental studies. PMID- 10791300 TI - Large-scale culture and preparation of sea urchin embryos for isolation of transcriptional regulatory proteins. PMID- 10791302 TI - Culture of avian embryos. PMID- 10791301 TI - The chick embryo as a model system for analyzing mechanisms of development. PMID- 10791303 TI - Exo ovo culture of avian embryos. PMID- 10791304 TI - Culture of preimplantation mouse embryos. PMID- 10791305 TI - In vitro culture of rodent embryos during the early postimplantation period. PMID- 10791306 TI - Cryopreservation of mouse embryos. PMID- 10791307 TI - Studying head and brain development in Drosophila. PMID- 10791308 TI - Bioassays of inductive interactions in amphibian development. PMID- 10791309 TI - Gastrulation and early mesodermal patterning in vertebrates. PMID- 10791310 TI - Craniofacial development of avian and rodent embryos. PMID- 10791311 TI - Examination of the axial skeleton of fetal rodents. PMID- 10791312 TI - Cardiac morphogenesis and dysmorphogenesis. An immunohistochemical approach. PMID- 10791314 TI - Confocal microscopy of live Xenopus oocytes, eggs, and embryos. PMID- 10791313 TI - Application of plastic embedding for sectioning whole-mount immunostained early vertebrate embryos. PMID- 10791315 TI - Whole-mount immunolabeling of embryos by microinjection. Increased detection levels of extracellular and cell surface epitopes. PMID- 10791317 TI - Embryo/fetal topographical analysis by fluorescence microscopy and confocal laser scanning microscopy. PMID- 10791316 TI - Confocal laser scanning microscopy of morphology and apoptosis in organogenesis stage mouse embryos. PMID- 10791318 TI - Magnetic resonance imaging analysis of embryos. PMID- 10791319 TI - Optical coherence tomography imaging in developmental biology. PMID- 10791320 TI - Ultrasound backscatter microscopy of mouse embryos. PMID- 10791321 TI - Use of Doppler echocardiography to monitor embryonic mouse heart function. PMID- 10791322 TI - Calcium imaging in cell-cell signaling. PMID- 10791323 TI - Acquisition, display, and analysis of digital three-dimensional time-lapse (four dimensional) data sets using free software applications. PMID- 10791324 TI - Cell lineage analysis. Applications of green fluorescent protein. PMID- 10791325 TI - Cell lineage analysis. X-inactivation mosaics. PMID- 10791326 TI - Retroviral cell lineage analysis in the developing chick heart. PMID- 10791327 TI - Dynamic labeling techniques for fate mapping, testing cell commitment, and following living cells in avian embryos. PMID- 10791328 TI - Cell lineage analysis. Videomicroscopy techniques. PMID- 10791330 TI - Photoactivatable (caged) fluorescein as a cell tracer for fate mapping in the zebrafish embryo. PMID- 10791329 TI - Cell lineage analysis in Xenopus embryos. PMID- 10791331 TI - Carboxyfluorescein as a marker at both light and electron microscope levels to follow cell lineage in the embryo. PMID- 10791332 TI - Transplantation chimeras. Use in analyzing mechanisms of avian development. PMID- 10791333 TI - Interspecific chimeras in avian embryos. PMID- 10791334 TI - Quail-chick transplantation in the embryonic limb bud. PMID- 10791335 TI - Mouse chimeras and the analysis of development. PMID- 10791336 TI - Cell grafting and fate mapping of the early-somite-stage mouse embryo. PMID- 10791337 TI - Interspecific chimeras. Transplantation of neural crest between mouse and chick embryos. PMID- 10791338 TI - Interspecific mouse-chick chimeras. PMID- 10791340 TI - Local application of bone morphogenic protein on developing chick embryos using fibrous glass matrix as a carrier. PMID- 10791339 TI - Mosaic analysis in Caenorhabditis elegans. PMID- 10791341 TI - Laser ablation and fate mapping. PMID- 10791342 TI - Photoablation of cells expressing beta-galactosidase. PMID- 10791343 TI - Exo utero surgery. PMID- 10791344 TI - Methods for constructing and producing retroviral vectors. PMID- 10791345 TI - Retroviral gene transduction in limb bud micromass cultures. PMID- 10791346 TI - Construction of adenoviral vectors. PMID- 10791347 TI - Application of adenoviral vectors. Analysis of eye development. PMID- 10791348 TI - The application of adenoviral vectors in the study of mammalian cardiovascular development. PMID- 10791349 TI - Liver tests. Simple blood tests can reveal a lot. PMID- 10791351 TI - Hysterectomy may improve sexual function for some women. PMID- 10791350 TI - Health tips. Care of an incision. PMID- 10791352 TI - A daily walk may help prevent diabetes. PMID- 10791354 TI - Warfarin. Proper use of this common anticoagulant is important. PMID- 10791353 TI - Hip-area pain. Narrowing the location of pain helps diagnosis. PMID- 10791356 TI - I tend to feel cold a lot, but my husband is always comfortable. Why the difference? PMID- 10791355 TI - Bioterrorism. How real is the threat? PMID- 10791357 TI - My fingernails tend to split and are soft and thin. Is there anything I can do to make them stronger? PMID- 10791358 TI - Birth outcomes of immigrant women in the United States, France, and Belgium. AB - OBJECTIVES: To compare maternal characteristics and birth outcomes of Mexico-born and native-born mothers in the United States and those of North African mothers living in France and Belgium to French and Belgian nationals. METHODS: We examined information from single live birth certificates for 285,371 Mexico-born and 3,131,632 U.S.-born mothers (including 2,537,264 U.S.-born White mothers) in the United States, 4,623 North African and 103,345 Belgian mothers in Belgium, and a French national random sample consisting of 632 North African and 11,185 French mothers. The outcomes were mean birthweight, low birthweight, and preterm births. Differences between native/nationals and foreign-born mothers in each country were assessed in bivariate and multivariate analyses controlling for maternal risk factors. RESULTS: The adjusted odds for low birthweight were lower for immigrants than native/nationals by 32% in the United States, by 32% in Belgium, and by 30% in France. The adjusted odds for preterm births were lower for immigrants compared with native/nationals by 11% in the United States and by 23% in Belgium. In France, the odds for preterm births were comparable for immigrants and naturalized mothers. Infants of immigrant mothers also had higher mean birthweights in all three countries. CONCLUSION: Despite their disadvantaged status, Mexico-born and North African-born women residing in the United States, France, and Belgium show good birth outcomes. These cannot be explained solely by traditional risk factors. Protective factors and selective migration may offer further clues. PMID- 10791359 TI - Racial differences in the patterns of singleton preterm delivery in the 1988 National Maternal and Infant Health Survey. AB - OBJECTIVES: To determine if the association between race and preterm delivery would persist when preterm delivery was partitioned into two etiologic pathways. METHODS: We evaluated perinatal and obstetrical data from the 1988 National Maternal and Infant Health Survey and classified preterm delivery as spontaneous or medically indicated. Discrete proportional hazard models were fit to assess the risk of preterm delivery for Black women compared with White women adjusting for potential demographic and behavioral confounding variables. RESULTS: Preterm delivery occurred among 17.4% of Black births and 6.7% of White births with a Black versus White unadjusted hazard ratio (HR) of 2.8 (95% CI = 2.4-3.3). The adjusted HR for a medically indicated preterm delivery showed no racial difference in risk (HR = 1.0, 95% CI = 0.4-2.6). However, for spontaneous preterm delivery between 20 and 28 weeks gestation, the Black versus White adjusted hazard ratio (HR) was 4.9 (95% CI = 3.4-7.1). CONCLUSIONS: Although we found an increased unadjusted HR for preterm delivery among Black women compared with White women, the nearly fivefold increase in adjusted HR for the extremely preterm births and the absence of a difference for medically indicated preterm delivery was unexpected. Given the differences in the risks of preterm birth between Black and White women, we recommend to continue examining risk factors for preterm delivery after separating spontaneous from medically indicated preterm birth and subdividing preterm delivery by gestational age to shed light on the reasons for the racial disparity. PMID- 10791360 TI - The Pregnancy Risk Assessment Monitoring System (PRAMS): methods and 1996 response rates from 11 states. AB - OBJECTIVES: To determine if the Pregnancy Risk Assessment Monitoring System (PRAMS) is a unique and valuable MCH data source and an effective mechanism for states to collect MCH data, and to assess if recent changes in it have improved efficiency and flexibility. METHODS: Each component of the PRAMS methodology is described: sampling and stratification, data collection, questionnaire, and data management and weighting. To assess effectiveness, we calculated response rates, contact rates, cooperation rates, refusal rates, and questionnaire completion rates. Logistic regression was used to examine the relationship between maternal and infant characteristics and the likelihood of response. Four criteria were defined to measure improvement in PRAMS functioning. RESULTS: Overall response rates for the 11 states in 1996 ranged from 66% to 80%. Cooperation rates were high (85-99%), with contact rates somewhat lower (73-87%). Response rates were higher for women who were older, White, married, had more education, were first time mothers, and had a normal-birthweight infant. In all states, parity and education were the most consistent predictors of response, followed by marital status and race. Between 1988-1990 and 1996-1999, the number of states and areas participating in PRAMS increased from 6 to 23, response rates improved, and the time for a state to start data collection and to obtain a weighted dataset both decreased. CONCLUSIONS: PRAMS is a unique and valuable MCH data source. The mail/telephone methodology used in PRAMS is an effective means of reaching most women who have recently given birth in the 11 states examined; however, some population subgroups are not reached as well as others. The system has become more efficient and flexible over time and more states now participate. PMID- 10791362 TI - Effective MCH epidemiology in state health agencies: lessons from an evaluation of the Maternal and Child Health Epidemiology Program (MCHEP). AB - OBJECTIVES: The Maternal and Child Health Epidemiology Program (MCHEP), jointly sponsored by the Centers for Disease Control and Prevention (CDC) and Health Resources and Services Administration (HRSA), was evaluated in 1996-1997. As part of this evaluation, an effort was undertaken to identify components of effective MCH epidemiology, to examine their prevalence across participating states, and to assess differences with respect to these components between MCHEP and non-MCHEP states. METHODS: A case-study evaluation was undertaken in which nine states (five MCHEP and four non-MCHEP) rated themselves on a benchmark questionnaire and participated in interviews conducted during site visits. At the completion of the evaluation, 16 components of effective MCH epidemiology in state health agencies were identified. The nine states were rated by the evaluation team on each component. Ratings across all states and between MCHEP and non-MCHEP states were compared. RESULTS: There was a great deal of variability among the nine states with respect to the presence of the components of effective MCH epidemiology. Components on which the states appeared weakest overall were the presence of adequately trained personnel, the presence of adequate management information systems to support MCH programs, and whether the state health agency's epidemiologic unit understands the MCH planning cycle. States with an MCHEP assignee had a higher overall mean score than non-MCHEP states across all components. There were seven components on which the two groups of states differed. These include whether the MCH director is empowered in the state health agency, whether the state health agency has identified internal epidemiologic capacity building as a priority, and whether analytic leadership is available for MCH epidemiologic activities. CONCLUSIONS: Building and maintaining MCH epidemiologic capacity in state health agencies requires attention to a variety of factors. While the presence of an MCH epidemiologist is important, this is only one of many components that must be considered as both the federal and state governments seek to promote and institutionalize effective MCH epidemiology in state health agencies. PMID- 10791361 TI - Using Medicaid data to estimate state- and county-level prevalence of asthma among low-income children. AB - OBJECTIVES: Asthma is one of the most common illnesses among children, yet there is little reliable information on the number of children at the state and county level who are living with asthma. This study examines the prevalence of asthma among low-income children in North Carolina using Medicaid paid claims and enrollment data. METHODS: Claims paid by Medicaid during state fiscal year 1997 1998 with a diagnosis of asthma or for a prescription drug used to treat asthma are examined to estimate prevalence among children ages 0-14 years. Percentages of enrolled children with asthma are presented by age, race, and rural/urban residence, and the costs of asthma treatment are calculated. RESULTS: More than 12% of North Carolina children ages 0-14 years on Medicaid had an indication of asthma. Prevalence rates were found to be highest among younger children, some minority groups, and residents of rural areas. More than $23 million was paid by Medicaid during the fiscal year for asthma-related services for children ages 0 14 years. CONCLUSIONS: State Medicaid databases are a useful means of studying the prevalence of asthma and other health conditions in low-income populations. Strengths and weaknesses of the proposed methodology are discussed. Existing administrative data systems can provide quick updates of prevalence rates at the state and county level, enhancing the ability to study trends in illness over time. PMID- 10791363 TI - 1994-1996 U.S. singleton birth weight percentiles for gestational age by race, Hispanic origin, and gender. AB - OBJECTIVES: Establishing and comparing race, ethnic, and gender-specific birth weight percentiles for gestational age is requisite for investigating the determinants of variations in fetal growth. In this study, we calculate percentiles of birth weight for gestational age for the total 1994-1996 U.S. population and contrast these percentiles by racial/ethnic and gender groups. METHODS: Single live births to U.S. resident mothers were selected from the 1994 1996 U.S. Natality Files. After exclusions, 5,973,440 non-Hispanic Whites, 1,393,908 non-Hispanic African Americans, 1,683,333 Hispanics, 80,187 Native Americans, and 510,021 other racial/ethnic groups were used to calculate distribution percentiles of birth weight for each gestational age for which there were at least 50 cases to calculate the 50th percentile and 100 cases to calculate the 10th percentile. RESULTS: Fetal growth patterns among the four U.S. racial/ethnic groups varied markedly and, across the gestational age range, there was considerable oscillation in the relative ranking of any one group's birth weight percentile value in comparison to the others. Males had relatively higher birth weight percentile values than females. The proportion of infants with a birth weight value less than 1994-1996 U.S. population's 10th percentile value of birth weight for their corresponding gestational age was 7.87 for non-Hispanic Whites, 15.43 for non-Hispanic African Americans, 9.30 for Hispanics, and 8.81 for Native Americans. CONCLUSIONS: While the factors underlying trends and population subgroup differences in fetal growth are unclear, nutrition, smoking habits, health status, and maternal morbidity are possible precursors for part of the variations in patterns of fetal growth. As prenatal care has been touted as a means to reduce the risk of fetal growth restriction at term, assuring the availability and accessibility of comprehensive prenatal care services is viewed as an essential corollary in the effort to improve fetal growth patterns in the United States. PMID- 10791364 TI - Multiple deliveries in North Carolina: effects on birth outcomes. AB - OBJECTIVES: The purpose of this study is to examine the trends in multiple deliveries in North Carolina and assess their effect on the rates of low birth weight, fetal mortality, and infant mortality. METHODS: Using North Carolina vital statistics files, trends in multiple births, categorized by race, maternal age, and birth weight, were examined for the period 1980-1997. A partitioning method was used to estimate the contribution of maternal age distribution and age specific multiple birth rates to the overall increase in multiple births, and the contribution of the changing multiple birth rate to observed trends in low birth weight and fetal and infant mortality. RESULTS: Between 1980 and 1997, the state's multiple birth rate increased by 40%. Most of the increase was due to a rise in the age-specific multiple birth rates, rather than a shift in the maternal age distribution. The increase in the multiple birth rate accounted for a substantial proportion of the increase in low birth weight among Whites and Blacks. The rise in multiple births also hindered further declines in fetal and infant mortality during this time. CONCLUSIONS: Multiple births are an increasingly important contributor to perinatal outcomes, and warrant greater consideration in research aimed at evaluating trends in low birth weight and infant mortality. PMID- 10791365 TI - Validity of a food frequency questionnaire in assessing nutrient intakes of low income pregnant women. AB - OBJECTIVE: In 1989, a validation study of eight nutrients was performed on a modified food frequency questionnaire (FFQ) specifically designed for low-income pregnant women. The purpose of this study was to broaden the scope of the previous study by assessing the validity of the FFQ for 17 additional nutrients. METHODS: The Pregnancy Food Frequency Questionnaire (PFFQ) was administered to a sample of 295 low-income, pregnant women aged 14-43 years living in Massachusetts. A randomly selected subsample of 101 women who provided at least one diet recall and reported intake of less than 4,500 calories were included in this analysis. RESULTS: Mean intake of 25 nutrients as assessed by one administration of the PFFQ and up to three diet recalls collected over 1 month were similar. Unadjusted correlation coefficients between nutrient intake measured by diet recalls and the questionnaire ranged from .28 (carotene) to .61 (folate). After adjusting for energy intake the correlations ranged from .03 (B12) to .46 (folate). The correlations corrected for day-to-day variation were higher, ranging from .07 (B12) to .90 (zinc). The mean correlation was .47 and there were 54% over .40. CONCLUSIONS: A food frequency questionnaire for English speaking, low-income, pregnant women can provide maternal and child health practitioners and researchers a valid estimate of diet across a wide range of nutrients. PMID- 10791366 TI - Nursing as a priority. PMID- 10791367 TI - Brain natriuretic peptide as bridge to therapy for heart failure. PMID- 10791368 TI - Origins of tremor. PMID- 10791369 TI - Temozolomide for malignant brain tumours. PMID- 10791370 TI - Research on intergenerational transmission of violence: the next generation. PMID- 10791371 TI - Women, pregnancy, and varicose veins. PMID- 10791372 TI - Haematology for non-haematologists. PMID- 10791373 TI - Comparison of radiotherapy alone with radiotherapy plus hyperthermia in locally advanced pelvic tumours: a prospective, randomised, multicentre trial. Dutch Deep Hyperthermia Group. AB - BACKGROUND: Local-control rates after radiotherapy for locally advanced tumours of the bladder, cervix, and rectum are disappointing. We investigated the effect of adding hyperthermia to standard radiotherapy. METHODS: The study was a prospective, randomised, multicentre trial. 358 patients were enrolled from 1990 to 1996, in cancer centres in the Netherlands, who had bladder cancer stages T2, T3, or T4, NO, MO, cervical cancer stages IIB, IIIB, or IV, or rectal cancer stage M0-1 were assessed. Patients were randomly assigned radiotherapy (median total dose 65 Gy) alone (n=176) or radiotherapy plus hyperthermia (n=182). Our primary endpoints were complete response and duration of local control. We did the analysis by intention to treat. FINDINGS: Complete-response rates were 39% after radiotherapy and 55% after radiotherapy plus hyperthermia (p<0.001). The duration of local control was significantly longer with radiotherapy plus hyperthermia than with radiotherapy alone (p=0.04). Treatment effect did not differ significantly by tumour site, but the addition of hyperthermia seemed to be most important for cervical cancer, for which the complete-response rate with radiotherapy plus hyperthermia was 83% compared with 57% after radiotherapy alone (p=0.003). 3-year overall survival was 27% in the radiotherapy group and 51% in the radiotherapy plus hyperthermia group. For bladder cancer, an initial difference in local control disappeared during follow-up. INTERPRETATION: Hyperthermia in addition to standard radiotherapy may be especially useful in locally advanced cervical tumours. Studies of larger numbers of patients are needed for other pelvic tumour sites before practical recommendations can be made. PMID- 10791374 TI - Treatment of heart failure guided by plasma aminoterminal brain natriuretic peptide (N-BNP) concentrations. AB - BACKGROUND: There is currently no objective practical guide to intensity of drug treatment for individuals with heart failure. We hypothesised that pharmacotherapy guided by plasma concentrations of the cardiac peptide aminoterminal brain natriuretic peptide (N-BNP) would produce a superior outcome to empirical trial-based therapy dictated by clinical acumen. METHODS: 69 patients with impaired systolic function (left-ventricular ejection fraction <40%) and symptomatic heart failure (New York Heart Association class II-IV) were randomised to receive treatment guided by either plasma N-BNP concentration (BNP group) or standardised clinical assessment (clinical group). FINDINGS: During follow-up (minimum 6-months, median 9.5 months), there were fewer total cardiovascular events (death, hospital admission, or heart failure decompensation) in the BNP group than in the clinical group (19 vs 54, p=0.02). At 6 months, 27% of patients in the BNP group and 53% in the clinical group had experienced a first cardiovascular event (p=0.034). Changes in left-ventricular function, quality of life, renal function, and adverse events were similar in both groups. INTERPRETATION: N-BNP-guided treatment of heart failure reduced total cardiovascular events, and delayed time to first event compared with intensive clinically guided treatment. PMID- 10791375 TI - Time from HIV-1 seroconversion to AIDS and death before widespread use of highly active antiretroviral therapy: a collaborative re-analysis. Collaborative Group on AIDS Incubation and HIV Survival including the CASCADE EU Concerted Action. Concerted Action on SeroConversion to AIDS and Death in Europe. AB - BACKGROUND: We used data from Europe, North America, and Australia to assess the effect of exposure category on the AIDS incubation period and HIV-1 survival and whether the effect of age at seroconversion varies with exposure category and with time since seroconversion. METHODS: 38 studies of HIV-1-infected individuals whose dates of seroconversion could be reliably estimated were included in the analysis. Individual data on 13030 HIV-1-infected individuals from 15 countries were collated, checked, and analysed centrally. We calculated estimates of mortality and AIDS incidence rates and estimated the proportions of individuals surviving and developing AIDS at each year after seroconversion from the numbers of observed deaths or cases of AIDS and the corresponding person-years at risk. Analyses were adjusted for age at seroconversion, time since seroconversion, and other factors as appropriate. FINDINGS: Mortality and AIDS incidence increased strongly with time since seroconversion and age at seroconversion. Median survival varied from 12.5 years (95% CI 12.1-12.9) for those aged 15-24 years at seroconversion to 7.9 years (7.4-8.5) for those aged 45-54 years at seroconversion, whereas for development of AIDS the corresponding values were 11.0 years (10.7-11.7) and 7.7 years (7.1-8.6). There was no appreciable effect of exposure category on survival. For AIDS incidence, the exposure category effect that we noted was explained by the high incidence of Kaposi's sarcoma in those infected through sex between men. We estimated that among people aged 25-29 years at seroconversion 90% (89-91) and 60% (57-62) survived to 5 years and 10 years, respectively, after seroconversion, whereas 13% (12-15) and 46% (44-49), respectively, developed AIDS (excluding Kaposi's sarcoma). INTERPRETATION: Before widespread use of highly-active antiretroviral therapy (before 1996), time since seroconversion and age at seroconversion were the major determinants of survival and development of AIDS in Europe, North America, and Australia. PMID- 10791376 TI - Consequences of discharges from intensive care at night. AB - BACKGROUND: It is generally believed that pressure for beds on intensive-care units (ICUs) has increased in the UK. This study used discharge at night as a proxy measure to investigate pressure. METHODS: Night was defined in two ways: "out of office hours' from 2200 to 0659 h and "the early hours of the morning" from 0000 to 0459 h. The rate of discharge at night was compared for 21 295 adult admissions to 62 ICUs covering the period 1995-98 with 10806 admissions to 26 ICUs covering the period 1988-90. With data solely from 1995-98, the consequences of discharge at night and premature discharge were investigated. FINDINGS: Overall, 2269 (21.0%) admissions did not survive the ICU in 1988-90 compared with 4487 (21.1%) in 1995-98. Of ICU survivors, 2.7% were discharged at night (2200 0659 h) in 1988-90 compared with 6.0% in 1995-98. In 1995-98, night discharges (2200-0659 h) had a higher crude (odds ratio 1.46, 95% CI 1.18-1.80) and case-mix adjusted (1.33, 1.06-1.65) ultimate hospital mortality. Higher odds ratios were observed when the definition of night was 0000-0459 h. Premature discharge was commoner at night, 42.6% vs 5.0% and its importance was apparent when incorporated into the logistic-regression model (premature discharge 1.35, 1.10 1.65; night discharge 1.17, 0.92-1.49). INTERPRETATION: Night discharges from ICU are increasing in the UK. This practice is of concern because patients discharged at night fare significantly worse than those discharged during the day. Night discharges are more likely to be "premature" in the view of the clinicians involved. The implication of these results is that many hospitals have insufficient intensive-care beds. In deciding whether or not to invest more resources in intensive care we must, however, consider the cost-utility of this particular service compared with other ways that additional resources could be used. PMID- 10791377 TI - Compartmentalised inducible nitric-oxide synthase activity in septic shock. AB - BACKGROUND: Previous experimental studies support a role for inducible nitric oxide synthase (iNOS) in the pathogenesis of severe sepsis. The aim of the study was to characterise iNOS activity in different tissues in patients with septic shock. METHODS: 13 consecutive patients with septic shock caused by cellulitis were enrolled. Skin, muscle, fat, and artery samples were obtained from normal, inflamed, and putrescent areas to measure iNOS activity, and concentrations of tumour necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta). In two patients, iNOS activity was also assessed in peripheral blood mononuclear cells (PBMC) incubated with microorganisms causing the sepsis, or in macrophages isolated from suppurating peritoneal fluid incubated with IL-1beta. FINDINGS: Compared with normal and inflamed areas, iNOS activity was increased in putrescent areas for muscle (71-fold [95% CI 20-259] vs normal areas, 69-fold [19 246] vs inflamed areas; p<0.01 for each) and for fat (68-fold [23-199] and 49 fold [18-137], respectively; p<0.01), but not for skin. Compared with normal areas, putrescent areas of arteries showed increased iNOS expression (1280-fold [598-3153]; p<0.01). Compared with normal areas, TNFalpha and IL-1beta were increased in putrescent areas of arteries (223-fold and 41-fold, respectively; p<0.01 for each). PBMCs and tissue macrophages expressed iNOS. Plasma nitrite/nitrate concentrations inversely correlated with mean arterial pressure and systemic vascular resistance. INTERPRETATION: In human septic shock we found that iNOS activity is compartmentalised at the very site of infection and parallels expression of TNFalpha and IL-1beta. PBMCs and tissue macrophages can be a cellular source for iNOS. PMID- 10791378 TI - Coherence between low-frequency activation of the motor cortex and tremor in patients with essential tremor. AB - BACKGROUND: In healthy people, rhythmic activation of the motor cortex in the 15 30 Hz frequency range accompanies and contributes to voluntarily-generated postural contractions of contralateral muscle. In patients with Parkinson's disease, an abnormal low-frequency activation of the motor areas of the cortex occurs and has been directly linked to the characteristic 3-6 Hz rest tremor of this disease. We therefore investigated whether the motor cortex is involved in the transmission of the rhythmic motor drive responsible for generating essential tremor. METHODS: Non-invasive recordings of activity from the hand area of the motor cortex were made from six patients with essential tremor by magnetoencephalography. The recordings were made simultaneously with the electromyogram recorded from contralateral finger muscles during periods of postural tremor. A statistical spectral analysis was done to determine at which frequencies the two signals were correlated. FINDINGS: Spectral analysis of the electromyogram signals showed a significant low-frequency component at the frequency of the tremor bursts. However, there was no coherence between magnetoencephalogram and electromyogram recordings at the tremor frequency, indicating that no correlation existed between the tremor signal and low frequency activity recorded from the primary motor cortex in individuals with essential tremor. Coherence at frequencies higher than the tremor frequency was similar to that in healthy individuals performing voluntary postural contractions. INTERPRETATION: The absence of significant coherence between the magnetoencephalogram and electromyogram at tremor frequencies suggests that in essential tremor the tremor is imposed on the active muscle through descending pathways other than those originating in the primary motor cortex. These findings challenge the model widely used to explain the efficacy of neurosurgical treatment of essential tremor, are in contrast to those of previous studies of parkinsonian rest tremor, and highlight an important difference in the pathophysiology of essential and parkinsonian tremor. PMID- 10791379 TI - Reversible dementia due to two coexisting diseases. PMID- 10791380 TI - Association of venous thromboembolism and clozapine. AB - Data from the Swedish Adverse Reactions Advisory Committee suggest that use of clozapine is associated with venous thromboembolic complications. We summarise 12 cases of thromboembolism during clozapine treatment. In five cases the outcome was fatal. PMID- 10791381 TI - Diabetes-associated autoantibodies in aboriginal children. AB - Type-2 diabetes is increasing in aboriginal children and adolescents and must be distinguished from type-1 diabetes in this population. The absence of diabetes associated autoantibodies supports the clinical impression of type-2 diabetes in the affected members of this population. PMID- 10791382 TI - Fetal origins of reduced arterial distensibility in the donor twin in twin-twin transfusion syndrome. AB - Twin-twin transfusion syndrome permits investigation of vascular programming independent of genetic influence. Arterial distensibiity was lower in the donor twin during infancy, implying the intrauterine vascular remodelling might result in raised cardiac afterload and could influence later cardiovascular health. PMID- 10791383 TI - Survival after introduction of HAART in people with known duration of HIV-1 infection. The CASCADE Collaboration. Concerted Action on SeroConversion to AIDS and Death in Europe. AB - We compared survival from HIV-1 seroconversion before and during the periods of highly-active antiretroviral therapy (HAART). Survival expectations were substantially better for HIV-1-infected individuals in 1997-98 (HAART period) compared with those in 1986-96 (pre-HAART period). Estimated survival 10 years from seroconversion was also substantially better in the later period. PMID- 10791385 TI - Combined ultrasonographically guided drainage and methotrexate administration for treatment of endometriotic cysts. AB - We report an effective alternative for the management of endometriotic cysts with transabdominal drainage under ultrasonographic control, followed by injection of 30 mg of methotrexate. PMID- 10791384 TI - Response of glassy-cell carcinoma of the cervix to cisplatin, epirubicin, and mitomycin C. AB - Among patients with uterine cervical cancer, glassy-cell carcinoma is very rare. The disease occurs mainly in younger patients, is rapidly progressive, and has a poor prognosis. We describe a case of glassy-cell carcinoma of the uterine cervix, in which preoperative intra-arterial chemotherapy was surprisingly effective. PMID- 10791387 TI - "Milestone" in meningococcal research: N. meningitidis A genome sequenced. PMID- 10791386 TI - USA to improve care of children with ADHD. PMID- 10791388 TI - Suzanne Cory: combining individuality with teamwork. PMID- 10791389 TI - Abortion returns to haunt US presidential campaign. PMID- 10791390 TI - Concern over supermarket sales of drugs in Spain. PMID- 10791391 TI - Questioning of HIV theory of AIDS causes dismay in South Africa. PMID- 10791392 TI - Human-rights groups express alarm at prosecution of Turkish doctors. PMID- 10791393 TI - WHO calls for international public hearing on tobacco control. PMID- 10791394 TI - Red cells I: inherited anaemias. AB - Examination of the genetic mechanisms underlying the thalassaemias has led to a clearer understanding of the control of eukaryotic genes in general. Inherited disorders of haemoglobin synthesis are an important cause worldwide of morbidity and mortality, and place a large burden on patients, families, and ultimately communities. The haemoglobin disorders can be controlled, by counselling and prenatal diagnosis. Treatment is usually symptomatic, though bone-marrow transplantation for beta-thalassaemia may be successful in suitable patients. PMID- 10791395 TI - The taxanes: an update. AB - The taxanes are anticancer cytotoxics that stabilise cellular microtubules. Two members, paclitaxel and docetaxel have substantial activity. One or both agents are widely accepted as evidence-based components of therapy for advanced breast, lung, and ovarian carcinomas. Paclitaxel has recently been approved in the USA for the adjuvant treatment of early stage node-positive breast carcinoma. PMID- 10791396 TI - The antioxidant paradox. PMID- 10791397 TI - The HOPE study and diabetes. Heart Outcomes Prevention Evaluation. PMID- 10791398 TI - The HOPE study and diabetes. Heart Outcomes Prevention Evaluation. PMID- 10791399 TI - The HOPE study and diabetes. Heart Outcomes Prevention Evaluation. PMID- 10791400 TI - The HOPE study and diabetes. Heart Outcomes Prevention Evaluation. PMID- 10791402 TI - Discontinuation symptoms and psychotropic drugs. PMID- 10791401 TI - The HOPE study and diabetes. Heart Outcomes Prevention Evaluation. PMID- 10791403 TI - Control of meningococcal disease in west Africa. PMID- 10791404 TI - Control of meningococcal disease in west Africa. PMID- 10791405 TI - Heterogeneity of juvenile dermatomyositis. PMID- 10791406 TI - Interpretation of Dutch BOA trial. Dutch Bypass Oral anticoagulants or Aspirin study group. PMID- 10791407 TI - Nosocomial influenza infection. PMID- 10791408 TI - Nosocomial influenza infection. PMID- 10791409 TI - Is Japan losing the fight against tobacco? PMID- 10791410 TI - Is Japan losing the fight against tobacco? PMID- 10791411 TI - Homicide, novel antipsychotics, and non-compliance. PMID- 10791412 TI - The homocysteine question. PMID- 10791413 TI - Social dis-ease of collective irresponsibility. PMID- 10791414 TI - High-voltage electron microscopy in neurocytology. AB - The function of the living matter from biomolecules to the whole body of the organism is based on the structure. Consequently, structural information is essential for the understanding of the function of living organisms. The biological structures from biomolecules to cells and tissues are intimately related to each other, and changing their morphological, biochemical, and physiological properties dynamically according to the developmental and functional status of the organism. The molecular dynamics of living matter should be related to the function of whole body. We employed HVEM stereoscopy and morphometry for the purpose of combining the structural information at nanometer level with those of the micrometer level. Novel aspects of three-dimensional organization of neuronal and glial cell processes have been presented. This structural information together with morphometrical data could contribute to the elucidation of brain function. PMID- 10791415 TI - Morphology and structure of various phases at the bonding interface of Al/steel formed by explosive welding AB - The bonding interface of explosively-welded aluminium and steel in three explosive conditions have been investigated by using scanning electron microscopy, transmission electron microscopy, electron diffraction and electron probe microanalysis methods. The results show that all the interfaces have the shape of waves with curled front formed by process of superplasticity and some discontinuous reacted zones. They consist of amorphous and nano sized crystals and quasi-crystals as well as the compounds such as AlFe, Al2Fe, Al3Fe and Al6Fe with various shapes. The basal steel crystal near the interface has structure of martensite and perlite crystals which are deformed by the process of superplasticity. The size of reacted zone becomes large with increasing amount of explosive charge powder and separation of the driver Al plate from the basal steel plate. PMID- 10791416 TI - Amplitude correction in image deconvolution for determining crystal defects at atomic level AB - A method is proposed to improve the image deconvolution technique based on the weak-phase object approximation for determining the crystal defects at atomic level. Originally, both the amplitude and phase modulation caused by the contrast transfer function (CTF) was removed for all reflections in the diffractogram of the image and then the deconvoluted image was obtained by carrying out the inverse FT. In this paper after removing the CTF modulation the amplitudes of reflections are further corrected by constraining the integral amplitudes to be equal to the corresponding structure factor amplitudes of perfect crystals. [110] high-resolution electron microscope images were simulated for a structure model of Si crystal containing the 60 degrees dislocation with different thickness. A comparison between deconvoluted images with the amplitudes corrected by the present method and those uncorrected is given to show the effectiveness of the method. The amplitude-crystal thickness dependence for images simulated close to the Scherzer defocus condition has been analysed and the result serves as an empirical basis of the method. The validity and limitation of the method are discussed. PMID- 10791417 TI - Crystal structure of the low-temperature phase of beta Cu1.75Se analysed by electron diffraction AB - The beta phase of Cu1.75Se has an anti-fluorite structure, where one in eight tetrahedral copper sites are vacant. Powder X-ray diffraction measurements on Cu1.75Se showed that the beta phase transformed to a two-phase mixture of (alpha + beta) at approximately 250 K, and then the beta phase changed to a new phase at approximately 180 K. Powder X-ray and electron diffraction measurements revealed that the low-temperature phase of the beta phase has a superstructure of (2alpha(fcc) x 2alpha(fcc) x 2alpha(fcc)) type, where alpha(fcc) is the lattice parameter of the original beta phase. The superstructure was interpreted to originate from the ordering copper vacancies. The new phase was referred to as the beta' phase. PMID- 10791418 TI - High precision phase-shifting electron holography AB - Today's information-oriented society requires high density and high quality magnetic recording media. The quantitative observation of fine magnetic structures by electron holography is greatly anticipated in the development of such new recording materials. However, the magnetic fields around particles <50 nm have not been observed, because the fields are too weak to observe in the usual way. Here we present a highly precise phase measurement technique: improved phase-shifting electron holography. Using this method, the electric field around a charged polystyrene latex particle (100 nm in diameter) and the magnetic field around iron particles (30 nm in diameter) are observed precisely. A precision of the reconstructed phase image of 2pi/300 rad is achieved in the image of the latex particle. PMID- 10791419 TI - Digital high-resolution electron microscopy of atomic disordering in YB56 AB - Despite of the difficulty in determining the atomic arrangement of light elements and doping atoms in the solid clusters, it is believed that the high-resolution images should have 'some' information on disordering, ordering, and doping atomic positions in these materials. In the present work, the information for disordering of atomic positions in YB56 was obtained from digital high-resolution images. The arrangement of Y atoms was also directly imaged and quantified. The present work indicates that the atomic detection by digital high-resolution images is very useful for evaluation of local structure in the ceramic materials with light elements such as boron. PMID- 10791420 TI - A new method for dynamical HOLZ calculations AB - An exact scheme for dynamical HOLZ calculations has been developed with a matrix formula based on the modified Bethe's theory and current flow without the small angle approximation. The HOLZ intensity must be calculated with the current flow for electron beams. For a quantitative intensity calculation at large illumination, not only the current flow in diffracted beams from an exit surface but also the current flow in a primary beam plays an important role. PMID- 10791421 TI - Bloch waves and weak-beam imaging of crystals AB - The influence of the number of diffracted beams on weak-beam contrast simulations of thickness contour lines and dislocation images is investigated. For large deviation parameters s(g)-->thickness contour lines from two-beam simulations are similar to those from many-beam simulations. In many-beam simulations of wedge shaped bent samples extra thickness contour lines appear at locations with ((g)- >, 3(g)-->). These extra lines occur between the imaging condition ((g)-->, -(g)- >) and ((g)-->, 3(g)-->). Therefore, in the case of a more symmetrical imaging condition many-beam simulations are mandatory. In bent samples the contributions of different Bloch waves to weak-beam images change as a function of the imaging conditions ((g)-->, x(g)-->). Near ((g)-->, 3.5(g)-->) two Bloch waves dominate. In the case of x <3 two other Bloch waves with different wavelengths are most important for the image contrast. The 'classical' ((g)-->, 3(g)-->) weak-beam condition is not suitable to determine signs and magnitudes of Burgers vectors from terminating thickness contour lines. Higher deviation parameters s(g)--> are necessary, especially for dense dislocation arrangements. PMID- 10791422 TI - On the grain boundary segregation of Sn in indium-tin-oxide thin films AB - The determination of Sn segregation by means of electron energy loss spectroscopy and X-ray photoelectron spectroscopy has been attempted on indium-tin-oxide thin films prepared by various processes: (a) as-deposited at room temperature; (b) after annealing at 533 K in vacuum; (c) after annealing in air; and (d) as deposited at 473 K. The occurrence of Sn segregation both at the surface and at the grain boundary was proved. The degree of segregation detected, varied depending on the preparation process of the films. PMID- 10791424 TI - TEM specimen preparation of a phase-change optical disk AB - It has been popular to use transmission electron microscopy (TEM) observation for investigating the microstructure of a phase-change optical disk. In the present work, a new method to prepare a plan-view TEM sample from a disk has been developed. In this method, a copper mesh is placed on a specific area of interest in the disk in advance and then the material is thinned down. By employing this procedure, it becomes possible for the first time to obtain foils that contain the specific area. Furthermore, an advanced method to prepare a cross-sectional TEM sample has also been developed, in which elimination of the polymer substrate is followed by ion milling. With this method, it is possible to prepare cross sectional foils for high-resolution and analytical electron microscopy observations. PMID- 10791423 TI - Electron microscopy analysis of the boundary layer structure of SrTiO3 semiconducting ceramic AB - In a boundary layer (BL) semiconducting SrTiO3-based ceramic condenser, the BL structure has been investigated using high-resolution field-emission scanning electron microscopy (FE-SEM) and field-emission (scanning) transmission electron microscopy (FE-(S)TEM). In an initial TEM observation, a double layered structure was observed at the grain boundary region. It consisted of a grain boundary (second phase) and a pair of the metal diffusion layers of up to several nanometres in width across the grain boundary where the change of the crystal lattice distance was undetected by the high-resolution TEM image. A facet structure was often observed on the grain boundaries. It was particularly formed on (020) plane of the grain crystal. High resolution SEM showed a jagged striped structure on the surface of the bulk material and on the inside grain as revealed by fracture. Using the similarity in shape and size, it can be identified to correspond to the facet boundary structure. Its formation mechanism can be explained as that during the reoxidization process when the oxide flux of the mixture of Bi2O3, PbO and CuO, painted on the bulk material, surfaces migrates into the ceramic along the grain boundary. The oxide corrodes the grain surfaces including the bulk surfaces. This corrosion particularly occurs on (020) plane of the grain so that the facet structure is produced. In this paper, by using the atomic scale high angle annular dark field STEM, it has been determined that Bi atoms preferentially replace Sr atoms on (020) in the diffusion layers. The atom position displacement was also detected at the grain surfaces and this altered atomic assignment can be determined as an origin of production of Sr2Bi4Ti5O18 at the grain boundary. Also, it was observed that the layer width of the metal diffusion layers was often different between the both grains and changed locally so that the ribbon of the diffusion layers meandered around the straight grain boundary. Its possible mechanism is also proposed. PMID- 10791425 TI - Characterization of lattice defects in strontium titanate single crystals by X ray topography and transmission electron microscopy AB - Lattice defects in SrTiO3 single crystals were characterized by X-ray topography and transmission electron microscopy. We examined two groups of crystals whose lapped faces were (001) and (011), respectively. After taking X-ray topographs, crystals which included relatively many defects were chosen for detailed investigation by transmission electron microscopy, which gave the following results: (i) some subgrain boundaries observed by X-ray topography were small angle tilt boundaries; and (ii) many dislocations were found in the region where thick line contrast was observed in X-ray topographs. Most of them had <100> type Burgers vectors. PMID- 10791426 TI - Microstructures in CoPtC magnetic thin films studied by superpositioning of micro electron diffraction AB - Cross-sectional transmission electron microscopy observation of CoPtC thin films showed that 10 nm sized ultrafine particles of CoPt typically were elongated along the substrate normal. Analysis of the superposition of 40 micro-electron diffraction patterns showed that there was no preferred crystal orientation of CoPt particles. This superpositioning technique can be applied to thin films, whose X-ray diffraction analysis is difficult due to the small size of the crystals. PMID- 10791427 TI - Image contrast of ultra HVEM and future ultra HREM AB - Review of the measurements and interpretation of energy dependence of extinction distance and transmissive power of electrons, which have appeared in the electron microscope images of crystals at the voltages ranging from 50 to 3000 kV, is presented. Applications of two beam dynamical theory of electron diffraction considering absorption effect of electrons made clear the importance of relativistic effect of the mass of electron to the variation of extinction distance and transmissive power at higher voltage and noted that they are proportional to (v/c) and (v/c)2 respectively, where v and c are the velocity of illuminating electrons and light. With elevating accelerating voltage of illuminating electrons, it was noted that many diffracted waves are excited and thus, for understanding of the image contrast and the improvement of transmissive power of electrons, many beam dynamical theory considering absorption of electron waves has to be applied. Detailed analysis of the excitation and absorption of component Bloch waves was carried out and flux (electron current) flow of Bloch waves and total waves was deduced. Improvements of image contrast and visibility of the imperfection in crystalline materials both in thin and thick regions of ultra-high voltages were realized by using the multi-beam imaging (MBI) method. In MBI of very thick crystals, it was noted that most of the inelastically scattered electrons which appeared in the background of the diffraction patterns contribute to the image contrast by the successive elastic scattering after inelastic scattering. It was noted that the improvement of maximum usable thickness at high voltage is highly dependent on the contribution of inelastically scattered electrons. The calculated flux flows of total electrons of various materials in various orientations were calculated for not only ultra high voltages but low voltages such as 100 kV. It was suggested that the intensity distribution of such flux flow will be recorded by the ultra HREM in the near future and will become useful to identify types of atoms, such as the interface consisting of different kinds of atoms. PMID- 10791429 TI - Practical SEM system based on the montage technique applicable to ultralow magnification observation, while maintaining original functions AB - A newly developed SEM system has been utilized for obtaining ultralow magnification SEM images. It is a successful combination of the modern SEM equipped with a motor drive stage fully controlled with PC and digital image processing techniques for automatic montage. In order to accomplish a practical system, several problems peculiar to the field of SEM, i.e. raster rotation, peripheral distortion and charging effects, are discussed and solved. The function of ultralow-magnification (whole area) observation is important during a scanning electron microscopy session. PMID- 10791428 TI - The discovery of the division apparatus of plastids and mitochondria. AB - Mitochondria and plastids contain distinct genomes and multiply by binary division of existing organelles. Mitochondrial and plastid division can be clearly separated into two main events: division of the organelle nuclei (nucleoids), and subsequent division of the rest of the organelles, the process of organellokinesis. Organellokinesis makes use of organelle dividing apparatuses such as plastid-dividing ring (PD ring) and mitochondrion-dividing ring (MD ring). The plastid-dividing apparatus (PD apparatus) is composed of three electron-dense rings (the outer, middle and inner), while the mitochondrion dividing apparatus (MD apparatus) is a pair of electron-dense rings in cytoplasm and inner ring in the mitochondrial matrix. The behaviour of both the PD and MD apparatuses throughout organelle division in Cyanidioschyzon merolae has been studied in detail by electron microscopy. When cells enter mitosis, the inner PD ring forms first, followed by the outer and middle rings and finally the MD rings. The PD rings begin to contract before the MD rings. However, the MD rings start to contract at about 4 times the speed of the PD rings and catch up to the PD rings. The cross-sectional areas of both the outer PD and MD rings increase as contraction in the plane of division progress. This suggests that the outer rings of organelle dividing apparatuses (OD apparatus) provide the motive force for contraction. FtsZ protein is located on the bacterial contractile ring at the equator of dividing bacteria, and controls bacterial division. Since FtsZ contains a tubulin motif, and host eukaryotic organisms and chloroplasts evolved from bacteria, there is debate whether that tubulins found in the cytoskeleton and the inner or outer PD ring evolved from FtsZ protein during eukaryogenesis. PMID- 10791430 TI - Electrostatic Russian quadruplet with high demagnification and minimum influence of geometric aberrations AB - Based on a combined analytical and numerical modelling, a high-quality probe forming system is proposed and developed. The Matrizant method is applied to minimize the beam spot size for a fixed beam current. For quadrupoles consisting of four metallic rods, the electric field inside of the quadruplet is computed using an accurate version of the boundary element method. The influence of the rod diameter on the beam spot size is studied for different emittances. The sensitivity of the system to energy spread and mechanical defects is also investigated. PMID- 10791431 TI - Atomic displacements in the modulated structure of Bi2Sr2(Ca(1 x)Pr(x))Cu2O(8+delta) and their effect on HRTEM reverse contrast image AB - The amplitude of transverse atomic displacement wave along the direction of incident electron beam exerts a significant effect on the brightness of spots in a high-resolution transmission electron microscope (HRTEM) reverse contrast image. The bright spot is associated with the projection of the modulated atomic chain along the beam direction. In general, the spot brightness is roughly inversely proportional to the amplitude of the displacement wave in an appropriate region of defocus, specimen thickness and amplitude. Monotonic decrease in the spot brightness with increasing amplitude can be attributed to the decrease in the degree of interference between scattered electrons. A complicated domain configuration has been found by the analysis of the spot brightness modulation observed in a [0 0 1] HRTEM reverse contrast image. PMID- 10791432 TI - Static capacitance contrast of LSI covered with an insulator film in low accelerating voltage scanning electron microscope AB - A new image contrast is reported for LSIs covered with an insulator film in a low accelerating voltage scanning electron microscope. The surface region above the conducting lines is often observed brighter than that without conducting lines. This contrast is quasi-stationarily observed contrary to well-known capacitive coupled voltage contrast, and is called static capacitance contrast. The optimum irradiation conditions for the maximum image contrast is studied and its mechanism is discussed. PMID- 10791433 TI - Plan-view imaging of oxygen-induced reconstruction on Ag(110) surface. I. The possibility of imaging surface oxygen AB - Surface reconstruction on Ag(110) induced by oxygen adsorption was studied by the plan-view imaging technique of high-resolution electron microscopy (HREM). Systematic multislice simulations were carried out to find the optimal experimental conditions for imaging the surface oxygen. It was found theoretically that there exist a series of characteristic values of objective lens defoci and sample thicknesses under which the surface and the bulk can be imaged clearly and simultaneously. These optimal imaging conditions were used to interpret the experimental micrographs. While the basic features of the surface reconstruction of O/Ag(110) in the HREM images are consistent with those revealed by scanning tunnelling microscopy (STM), there are some new features that have not been reported with STM. These phenomena give some evidence to the existence of subsurface oxygen. PMID- 10791434 TI - Plan-view imaging of oxygen-induced reconstruction on Ag(110) surface. II. Effect of high-energy electron thinning AB - The feasibility and the limitation of the 'high-energy electron thinning' method for the production of surface-science-grade samples in situ in the electron microscope are studied. Exploiting the electron beam supplied by high-voltage electron gun in electron microscope, this method can be readily realized. An obvious advantage of this method is that we can monitor the sample surface concurrently. However, this sample preparation method depends strongly on the sample material and the local environment within the electron microscope. Factors relating to the electron thinning speed are briefly discussed. PMID- 10791435 TI - Simulation of the effect of magnification and image-rotation variations on three dimensional reconstruction AB - Tilting a specimen may change the magnification and the image rotation for electron-microscopic images, because re-focusing with an objective lens is required to correct for the height variation of the tilted specimen. Computer simulation is performed to analyse the effect of variations in the magnification and the image rotation on the three-dimensional reconstruction in electron tomography. PMID- 10791436 TI - Morphological changes of capillaries in the rat soleus muscle following experimental tenotomy. AB - We examined the structural changes of capillaries in the rat soleus muscle 4, 7, 14, and 35 days after experimental limb tenotomy. In the soleus muscles after tenotomy, muscle fibres degenerated and some of them were destroyed; the muscle did not seem to recover until the 35th day. On the 14th day, some small muscle fibres, probably regenerating muscle fibres, started forming within the basal lamina tube and remained after necrosis of a pre-existing muscle fibre. Most capillaries at each stage were of the continuous type. However, about 10% of the capillaries around degenerated muscle fibres at days 4, 7 and 14 consisted of endothelial cells with a small number of fenestrae bridged by a single-layered diaphragm. On the 14th day, capillaries around small regenerating muscle fibres also often had a small number of fenestrations. Even on the 35th day, capillaries occasionally had fenestrations. Additionally, some of the fenestrated capillaries formed small pores at the fenestrated portion of the endothelial cells. The untreated muscles contained only continuous capillaries. These findings suggest that fenestrations in the endothelial cells may occur in intramuscular capillaries not only around degenerated muscle fibres but also around regenerating muscle fibres after tenotomy. PMID- 10791437 TI - Scanning electron microscopy subsequent to a combined treatment of NaOCl and EDTA in some non-collagenous calcified matrixes. AB - Using backscattered electron (BSE) imaging and scanning electron microscopy, subsequent to a combined treatment of sodium hypochlorite (NaOCl) and ethylenediamine tetra-acetic acid (EDTA) or only with EDTA etching, we observed some structures of non-collagenous calcified matrixes with the aim of revealing the correlation of deposition between calcification degree and organic amount. In human tooth enamel, the NaOCl-EDTA method eroded more intensively the hypocalcified prisms of enamel tufts containing a relatively large amount of EDTA insoluble organic matter than the hypercalcified normal prismatic enamel containing a small amount of the organic matter. Afibrillar cementum, one of the non-collagenous calcified tissues similar to the enamel, has been reported to consist of organic-rich and poor incremental lamellae. The BSE imaging showed an alternation pattern of hypocalcification and hypercalcification. The hypocalcified lamellae were retained by EDTA etching, while the hypercalcified lamellae showed a resistance against the NaOCl-EDTA method. In the non collagenous calcareous concretions of human pineal body, organic-rich and poor, and hyper- and hypocalcified incremental lamellae have been reported. The deposition pattern of calcification degree and organic amount was similar to that in afibrillar cementum, and the hypercalcified lamellae showed a resistance against the NaOCl-EDTA method. In conclusion, the high and the lower calcified regions of non-collagenous calcified matrixes contained smaller and larger amounts of EDTA-insoluble organic matter respectively. Moreover, scanning electron microscopy subsequent to the NaOCl-EDTA method corresponding to the BSE imaging clearly showed fine calcified structures compared with the BSE imaging. PMID- 10791438 TI - A re-examination of the cellular reticulum of fibroblast-like cells in the rat small intestine by scanning electron microscopy. AB - The organization and arrangement of fibroblast-like cells in the rat small intestine were re-examined by scanning electron microscopy after removal of the epithelium and underlying connective tissue components by HCl hydrolysis. In the villi, the fibroblast-like cells had numerous slender processes, and formed a dense and elaborate network like a sieve. It consisted of a large number of circular structures (circles) with various diameters ranging from 0.3 microm to 5 microm formed by the twining of slender processes. In the upper area of villi, numerous fragmental protrusions which were considered to be mainly parts of immune-related cells such as lymphocytes, eosinophils and macrophages extended from the circles. The cells around each tubular gland enclosed it like a basket. These findings suggest that in addition to the function as a skeleton for the villi and glands, the fibroblast-like cells in the upper area of villi may play an important role in regulating the migration of the immune-related cells between the epithelial layer and the underlying lamina propria by their cellular sieve like structure and contractile ability. PMID- 10791439 TI - Compression or expansion of morbidity--what does the future hold? PMID- 10791440 TI - Measuring quality of care. PMID- 10791441 TI - Caring for older hospital-at-home patients. PMID- 10791442 TI - Thiamine deficiency in elderly people. PMID- 10791443 TI - Treatment of Helicobacter pylori infection in elderly subjects. PMID- 10791444 TI - Is thiamine deficiency in elderly people related to age or co-morbidity? AB - OBJECTIVES: to compare erythrocyte thiamine pyrophosphate concentrations in elderly people with those in healthy younger people; to determine if any differences can be attributed to age or to co-morbidities. DESIGN: cross sectional and 3-year longitudinal surveys. SETTING: primary care. PATIENTS: 100 volunteer blood donors and 222 elderly people from a general practice register. MEASUREMENTS: thiamine pyrophosphate concentrations using high performance liquid chromatography; physical examination, medical and medication history; grip strength, body mass index and plasma albumin. RESULTS: the mean [95% confidence interval (CI)] thiamine pyrophosphate concentration was 152 nmol/l (147-158) in the elderly group and 224 (213-235) nmol/l in the younger group (P < 0.001). Ninety-six (43.4%) of the elderly subjects had thiamine pyrophosphate concentrations below the fifth percentile of the younger subjects (140 nmol/l). Over 3 years thiamine pyrophosphate concentrations fell in the elderly cohort by 20% (95% CI: 14.5-24.5%; P < 0.01). Thiamine pyrophosphate concentrations in 39 healthy older people were no different from those in elderly people with co morbidity but were significantly lower than those in the younger people. Elderly people with absent vibration sense in their feet had a lower thiamine pyrophosphate concentration than the rest of the group [129 (117-142)nmol/l compared with 156 (150-162)nmol/l; P < 0.01)]. Thiamine pyrophosphate concentrations were not related to prevalent diseases, common medications, body mass index, grip strength or plasma albumin. CONCLUSION: lower thiamine pyrophosphate concentrations in elderly people appear to be related more to age itself than to co-existent illnesses. PMID- 10791445 TI - A comparison of fluconazole oral suspension and amphotericin B oral suspension in older patients with oropharyngeal candidosis. Multicentre Study Group. AB - BACKGROUND: the optimum treatment for oropharyngeal candidosis, particularly in older patients, has not been established. Local treatment with nystatin and amphotericin B can be problematic. The oral suspension formulation of fluconazole may offer a good alternative to these conventional agents. OBJECTIVE: to compare the safety and efficacy of fluconazole oral suspension with amphotericin B oral suspension in the treatment of older patients with oropharyngeal candidosis. DESIGN: randomized open-label study. PATIENTS: three hundred and five patients, aged 62 or older, with at least one sign or symptom of oropharyngeal candidosis. METHODS: we evaluated patients for the signs and symptoms of candidosis before receiving the study drug and on days 4, 7 and 14. We assessed patients who were cured or improved after 7-14 days of treatment 2 weeks after the end of treatment (follow-up). We obtained specimens from buccal lesions for microscopic examination (baseline only) and culture at baseline and on days 7 and 14. Patients were evaluated for adverse events on days 4, 7 and 14. RESULTS: one hundred and fifty patients received fluconazole and 155 received amphotericin B. There were no statistically significant differences in clinical or mycological response between fluconazole and amphotericin B at the end of treatment or at follow-up. At the end of treatment, 122 (81%) of 150 fluconazole-treated and 135 (87%) of 155 amphotericin B-treated patients were clinically cured or improved. Mycological cure rates were 35% and 46% for fluconazole and amphotericin B, respectively. The symptoms of burning sensation and buccal pain resolved significantly sooner (P < 0.05) in fluconazole-treated patients. The presence of dentures did not affect the response to antifungal therapy. The incidence of adverse events was 46% in the fluconazole group and 50% in the amphotericin B group (not statistically significant). CONCLUSION: fluconazole oral suspension is a good therapeutic alternative to amphotericin B oral suspension in the treatment of older patients with oropharyngeal candidosis. PMID- 10791446 TI - Symptom severity and oesophageal chemosensitivity to acid in older and young patients with gastro-oesophageal reflux. AB - INTRODUCTION: elderly patients appear to have a more aggressive form of gastro oesophageal reflux disease than younger patients. Reduced pain perception with age is a possible underlying mechanism. AIMS: to compare the extent of oesophageal mucosal injury, acid exposure, symptom severity and perception thresholds for acid infusion in older (aged 60 or older) and younger patients with gastro-oesophageal reflux. METHODS: twenty-five younger and 23 older patients completed the study. We determined acid exposure and oesophageal mucosal injury by ambulatory 24-h oesophageal pH monitoring and upper endoscopy, respectively. We determined chemosensitivity by infusing 0.1 N hydrochloric acid into the mid-oesophagus for 10 min at 10 ml/min after a 2-min infusion of normal saline at a similar rate. We quantified acid perception thresholds by the lag time to initial typical symptom perception, intensity rating at the end of acid infusion and an acid perfusion sensitivity score, calculated from the fractional duration of symptom perception and intensity rating. RESULTS: mean percentage of total time with pH <4 was higher in the older (15.8+/-2.4) than in the younger patients (11.9+/-1.8; P = 0.18). Of the older group, 74% had erosive oesophagitis versus 64% in the younger group. Frequency of symptoms (heartburn, acid regurgitation and dysphagia) was lower in the elderly group. Older patients perceived heartburn and acid regurgitation as much less severe than younger patients (P < 0.05).Younger patients had a significantly shorter lag time to initial symptom perception (P < 0.05) and a higher sensory intensity rating (P < 0.08). The acid perfusion sensitivity score was significantly lower in the older group (P < 0.05). CONCLUSIONS: older patients with gastro-oesophageal reflux disease have reduced symptom severity for heartburn despite a tendency towards increased severity of oesophageal mucosal injury and acid exposure. Age-related reduction in chemosensitivity to acid is a possible underlying mechanism. PMID- 10791447 TI - Uptake of breast cancer screening in older women. AB - OBJECTIVES: to determine breast screening uptake in older women and to ascertain from previous non-attenders whether they would accept screening if invited. DESIGN: a random sample of older women randomly selected from three Family Health Service Authorities were interviewed in their homes. PARTICIPANTS: 1604 women aged 65 years and over living at home, a response rate of 94%. RESULTS: 120 respondents (8%) had previously been screened. Rates were higher among those who were married, separated or divorced than those who were single (P < 0.01). Of those who had not previously been screened, 742 (50%) reported that they would attend if invited. Age influenced potential attendance: 67% of those aged 65-69 would accept compared with 27% of those aged 80 and over (P < 0.0001). Future attenders were significantly more likely to belong to the upper social class and to be currently married, and were significantly less likely to be disabled or depressed, but more likely to be anxious (P < 0.05). CONCLUSIONS: it is unjustifiable to exclude women over 65 from breast screening on the basis of assumed low uptake rates. Certain categories of women--such as those who were physically disabled, depressed, single or from lower social classes--could be targeted to achieve maximum uptake rates. PMID- 10791448 TI - How great a burden does early discharge to hospital-at-home impose on carers? A randomized controlled trial. AB - OBJECTIVE: to assess the effects of an early discharge hospital-at-home scheme on self-reported carer strain and quality of life. DESIGN: a randomized controlled trial SETTING: Bristol, UK. SUBJECTS: 133 carers of patients receiving either early discharge from hospital to hospital-at-home (n = 93) care or usual hospital care and discharge (n = 40). OUTCOME MEASURES: modified 12-item Carer Strain Index, COOP-WONCA charts and EuroQol EQ-5D at 4 weeks and 3 months post randomization. RESULTS: the mean age of carers was 65 years; 56% were women. There were no marked differences between the groups in any of the outcomes used at either 4-week or 3-month follow-up. CONCLUSION: there was no evidence of increased self-reported burden imposed on carers of patients discharged early from hospital. Decisions on the implementation of hospital-at-home schemes should be influenced by considerations of cost and effectiveness rather than effects on carers. The effects on carers may, however, differ for other forms of home-based care. PMID- 10791449 TI - Healthy ageing is associated with reduced and delayed disability. AB - OBJECTIVE: to identify subgroups within the population with reduced or delayed disability during healthy ageing. DESIGN: a longitudinal, community-based study. SETTING: Dubbo, New South Wales, Australia. PARTICIPANTS: 2805 men and women 60 years and older, first examined in 1988-89. OUTCOME MEASURES: activities of daily living assessed serially every 2 years over 8 years (scored in the range 0-6, least to most impaired); scores related to subsequent hospital admissions and to demographic, clinical and psychosocial characteristics at baseline. RESULTS: 1973 men and women provided complete follow-up data. Mean disability score at entry was low at 0.18 and increased to 0.69 by the final survey. Those having three or more hospital admissions (40% of the sample) had minimum disability (disability score approximately 0.3) around 5 years earlier than those with fewer admissions. Those with dementia or other mental illness had the most severe disability (mean disability scores of 3.15 and 2.13 respectively), but their numbers were very small. Those with a stroke or respiratory illness were more numerous and they had major physical disability (mean disability scores of 1.44 and 1.32 respectively). In a regression model, the statistically significant baseline predictors of disability at the final survey were age, body mass index, use of anti hypertensive medication, history of stroke, depression score, peak expiratory flow and physical disability. CONCLUSIONS: the findings confirm reduced or delayed disability in older citizens requiring little or no hospitalization. Age, impaired peak expiratory flow and physical disability at study entry were most strongly predictive of disability, while stroke and respiratory illness were relatively common causes of severe disability. PMID- 10791450 TI - Socio-demographic differences in the onset and progression of disability in early old age: a longitudinal study. AB - OBJECTIVES: to analyse socio-demographic differences in the onset and progression of disability. DESIGN: analysis of a cohort of people aged 55-69 in 1988-9 and in 1994. SUBJECTS: a representative sample of 3543 adults. METHODS: we measured severity of disability at baseline and follow-up. We analysed variations in incidence and progression of disability by using logistic regression. RESULTS: Baseline severity of disability was similar for men and women but varied by age group, social class, educational qualifications and housing tenure. At follow-up, 36% had worse disability, 12% better and 53% the same as at baseline. Increased severity of disability and new incidence of disability were associated with lower socioeconomic status, baseline self-rated health status, age and gender. High initial levels of disability were associated with improvement at follow-up. CONCLUSION: disability can be dynamic, although deterioration is more usual than improvement. The reasons for the associations found between disability and socio economic status are unclear. PMID- 10791451 TI - Does treatment of constipation improve faecal incontinence in institutionalized elderly patients? AB - OBJECTIVE: to evaluate whether faecal incontinence can be improved by treatment of constipation in elderly patients with faecal incontinence associated with impairment of rectal emptying. DESIGN: a prospective randomized study with a 2 month follow-up. SETTING: five long-term care units. SUBJECTS: 206 patients with daily faecal incontinence associated with chronic rectal emptying impairments such as faecal impaction received either a single osmotic laxative (group I) or an osmotic agent along with a rectal stimulant and weekly enemas (group II). MEASUREMENTS: episodes of faecal incontinence and associated details of soiled laundry (used as indicators of the workload for caregivers). We performed periodic digital rectal examinations on group II patients to evaluate whether treatment resulted in complete and long-lasting rectal emptying. We compared data between groups and in group II between persistently constipated patients and patients with complete rectal emptying. RESULTS: the frequency of faecal incontinence did not significantly differ between the two groups. The 23 patients in group II who had complete rectal emptying had 35% fewer episodes of faecal incontinence and 42% fewer incidents of soiled laundry than the rest of the group. CONCLUSIONS: when long-lasting and complete rectal emptying is achieved by laxatives, the number of episodes of faecal incontinence as well as the workload for caregivers is reduced. PMID- 10791452 TI - Development of a minimum data set-based depression rating scale for use in nursing homes. AB - BACKGROUND: depression is common but under-diagnosed in nursing-home residents. There is a need for a standardized screening instrument which incorporates daily observations of nursing-home staff. AIM: to develop and validate a screening instrument for depression using items from the Minimum Data Set of the Resident Assessment Instrument. METHODS: we conducted semi-structured interviews with 108 residents from two nursing homes to obtain depression ratings using the 17-item Hamilton Depression Rating Scale and the Cornell Scale for Depression in Dementia. Nursing staff completed Minimum Data Set assessments. In a randomly assigned derivation sample (n = 81), we identified Minimum Data Set mood items that were correlated (P < 0.05) with Hamilton and Cornell ratings. These items were factored using an oblique rotation to yield five conceptually distinct factors. Using linear regression, each set of factored items was regressed against Hamilton and Cornell ratings to identify a core set of seven Minimum Data Set mood items which comprise the Minimum Data Set Depression Rating Scale. We then tested the performance of the Minimum Data Set Depression Rating Scale against accepted cut-offs and psychiatric diagnoses. RESULTS: a cutpoint score of 3 on the Minimum Data Set Depression Rating Scale maximized sensitivity (94% for Hamilton, 78% for Cornell) with minimal loss of specificity (72% for Hamilton, 77% for Cornell) when tested against cut-offs for mild to moderate depression in the derivation sample. Results were similar in the validation sample. When tested against diagnoses of major or non-major depression in a subset of 82 subjects, sensitivity was 91% and specificity was 69%. Performance compared favourably with the 15-item Geriatric Depression Scale. CONCLUSION: items from the Minimum Data Set can be organized to screen for depression in nursing-home residents. Further testing of the instrument is now needed. PMID- 10791453 TI - Quality of care from the perspective of elderly people: the QUOTE-elderly instrument. AB - BACKGROUND: patient views on the quality of care are usually assessed by means of patient satisfaction questionnaires. AIM: to develop an instrument that would: (i) produce data related to the expectations and experiences of noninstitutionalized elderly people, (ii) contain items that had been formulated in collaboration with elderly people, (iii) measure quality from the perspective of the users of health care services and (iv) produce data on generic quality aspects and quality aspects specifically related to the needs of elderly people. METHODS: we developed the instrument for measuring quality of care from the perspective of non-institutionalized elderly people (QUOTE-Elderly) by using a combination of qualitative and quantitative methods. We obtained empirical data on the opinions and experiences of 338 elderly people. We evaluated the taxonomy of the instrument, internal consistency of (sub)scales and the feasibility of the instrument using explorative and confirmative factor analyses and reliability analysis. RESULTS: using scale optimization, we produced a self-administered questionnaire on quality of health care from the perspective of elderly people. This contains scientific characteristics and provides specific information for practical quality-assurance policies. PMID- 10791454 TI - A question of competence. PMID- 10791455 TI - Age-related geriatric medicine or integrated medical care. PMID- 10791456 TI - Age-related geriatric medicine or integrated medical care. PMID- 10791457 TI - Randomized trials and the use of anticoagulants for atrial fibrillation. PMID- 10791458 TI - Won't drink? Can't drink. PMID- 10791460 TI - The other side of hip protectors. PMID- 10791459 TI - Additional risk factors in atrial fibrillation patients not receiving warfarin. PMID- 10791461 TI - Seasonal effects on cardiovascular mortality in older patients. PMID- 10791463 TI - Hip fracture in the 1930s. PMID- 10791462 TI - Metformin and ageing diabetic patients. PMID- 10791464 TI - Committee's concerns over feedingstuffs regulations. PMID- 10791465 TI - Cases of sheep scab widely encountered during February. PMID- 10791466 TI - Scrapie surveillance in Great Britain: results of an abattoir survey, 1997/98. AB - A randomised sample of 2,809 apparently healthy sheep, 55 per cent of them less than 15 months of age, which were slaughtered for human consumption at abattoirs in Great Britain in 1997/98, was taken to establish the prevalence of scrapie infection. The medulla oblongata of each sheep was examined histopathologically at the level of the obex, and fresh brain tissue was examined for scrapie associated fibrils (SAF) to establish whether there was evidence of scrapie. In addition, histological sections of the medulla from 500 of the sheep were immunostained with an antiserum to PrP, and the same technique was also applied to any animal found positive or inconclusive by the histological or SAF examinations. Any sheep which was positive by any of these diagnostic methods was also examined by Western immunoblotting, for the detection of the disease specific protein PrP(Sc). A total of 2,798 sheep (99.6 per cent) were negative by all the methods applied. Ten animals were SAF-positive but negative by all the other methods, and in one animal there was immunohistochemical staining which could not be interpreted unequivocally as disease-specific. A mathematical model was used to estimate the prevalence of scrapie infection in the national slaughtered sheep population which would be consistent with these results. By this model, the absence of unequivocally substantiated cases of scrapie in the sample was consistent with a prevalence of infection in the slaughter population of up to 11 per cent. PMID- 10791467 TI - Repeated oestrus synchronisation of beef cows with progesterone implants and the effects of a gonadotrophin-releasing hormone agonist at implant insertion. AB - A group of 97 spring-calving beef cows were initially oestrus synchronised with controlled internal drug release (CIDR) intravaginal progesterone implants inserted for nine days and a prostaglandin injection on day 7. Approximately half the cows were given 10 microg buserelin when the implants were inserted, and they all received a single fixed-time artificial insemination (AI) 56 hours after the withdrawal of the implants. The overall pregnancy rate to the first synchronised AI was 55 per cent, the buserelin-treated cows having a pregnancy rate of 63 per cent compared with 47 per cent in the untreated cows (P>0.05). Sixteen days after the first synchronised AI all the cows were re-implanted with used CIDR implants which were removed five days later, and the cows received a second synchronised AI on days 23 to 24. Cows which received the second AI were implanted with new CIDR devices 16 days later and these were removed after five days and the non pregnant cows received a third synchronised AI. The pregnancy rates to the second and third synchronised services were 74 per cent and 75 per cent, respectively. PMID- 10791469 TI - Immunohistochemical study of hormonal receptors and cell proliferation in normal canine mammary glands and spontaneous mammary tumours. AB - The expression of hormone receptors and their relationship to cell proliferation in six samples of normal canine mammary tissue, and 11 benign and 10 malignant mammary neoplasms from female dogs were assessed by immunohistochemistry in formalin-fixed paraffin-embedded samples, by using monoclonal antibodies against progesterone and oestrogen receptors, and nuclear antigen Ki-67 (MIB-1). Malignant tumours negative for progesterone receptors proliferated at higher rates than progesterone receptor-positive tumours, suggesting that the progression towards malignancy in spontaneous mammary tumours is accompanied by a decrease in hormonal steroid dependency. Only one malignant tumour was positive for oestrogen receptors. PMID- 10791468 TI - Prevalence of Brachyspira species isolated from diarrhoeic pigs in Brazil. AB - Pathogenic intestinal spirochaetes of pigs include Brachyspira (formerly Serpulina) hyodysenteriae, the cause of swine dysentery, and Brachyspira pilosicoli, the cause of porcine colonic spirochetosis (PCS). The purpose of this study was to assess the relative importance of Brachyspira species in diarrhoeal disease of growing pigs on farms in southern Brazil. The intensity and pattern of haemolysis, the production of indole and the hydrolysis of hippurate by reference and field porcine intestinal spirochaetes were compared with 16S-ribosomal RNA (mRNA)- and 23S-rRNA-based polymerase chain reaction assays for the identification of B hyodysenteriae and B pilosicoli. Between July and October 1998, 206 rectal swabs were taken from pigs on 17 farms with a history of diarrhoea developing within 30 days after they had been moved from nursery to growing facilities. Of 49 beta-haemolytic spirochaetes that were cultured, 29 (59.2 per cent) were grown in pure culture for phenotypic and genotypic characterisation, leaving 20 untyped. Of the 29 typed isolates, eight isolates obtained from six farms were identified as B hyodysenteriae, and 15 isolates obtained from seven other farms were identified as B pilosicoli; the remaining six isolates were identified as weakly beta-haemolytic commensal spirochaetes. There was complete agreement between the results of the phenotypic and genotypic analyses. PMID- 10791470 TI - Bovine tuberculosis in domestic cats. PMID- 10791471 TI - NVQ training scheme. PMID- 10791472 TI - NVQ training scheme. PMID- 10791473 TI - Plesiomonas shigelloides--an uncommon cause of diarrhoea in cats? PMID- 10791474 TI - Exotic pets. PMID- 10791475 TI - Comparison of eating attitudes and behaviors among adolescent and young women with type 1 diabetes mellitus and phenylketonuria. AB - The purpose of this study was to assess the eating attitudes and behaviors associated with two chronic diseases that have strong dietary treatment components: type 1 diabetes mellitus and phenylketonuria (PKU). Participants consisted of female campers and staff members who were attending one of two summer camps that specialize in the care of females with type 1 diabetes mellitus (N = 54) and PKU (N = 30). Eating attitudes and behaviors, psychological adjustment, and disease-specific knowledge were assessed using standardized and nonstandardized self-report questionnaires. There was no overall difference in the presence of disordered eating symptomatology between those with diabetes and those with PKU. However, differences in patterns of eating attitudes and behaviors were observed. The data suggest that living with chronic diseases which are treated with dietary management may adversely affect eating attitudes and behaviors and may increase susceptibility to the development of eating disturbances. PMID- 10791476 TI - Young women with physical disabilities: risk factors for symptoms of eating disorders. AB - Women with disabilities are at risk for poor psychological adjustment and unhealthy weight-control practices. This study was conducted to determine the prevalence of symptoms of eating disorders in a clinic-based sample of women who had two disabling conditions and to identify condition-related risk factors that were linked to these symptoms. A cross-sectional survey of 71 women (mean age = 23 yr) with spina bifida or rheumatologically related illnesses was conducted to assess the symptoms of eating disorders, condition-related characteristics, and weight-control practices. Symptoms of eating disorders were measured by the Eating Disorders Inventory (EDI). Eight percent of the respondents reported a sufficient number of symptoms of eating disorders to indicate a possible clinical disorder. More than 20% of the respondents scored at or above the clinical cut point on at least one of the eight EDI subscales. Selected condition-related characteristics (multiple conditions, condition-affected driving, and uncertainty of illness course) were associated with EDI scores after adjusting for demographic variables, family factors, and weight-control practices. PMID- 10791477 TI - Mothers' and fathers' perceptions of three year olds' attachment behavior. AB - This investigation reports on the antecedents and cross-sectional predictors of young children's attachment security based on Q-sort methodology, with particular consideration of the convergence of parental reports. Mothers' and fathers' ratings of child temperament and child behavior problems, as well as each parent's construction of the marital relationship and his/her affective state, were examined in relation to Q-sort assessments of mother-child and father-child attachment security. Generally, a moderate degree of convergence between maternal and paternal perceptions of attachment security was found; this association was stronger than the correlations reported based on studies using the Strange Situation procedure at 12 to 18 months of age. Antecedent relationships between ratings of parental affective symptoms, as well as ratings of marital quality and security of attachment, were stronger for fathers than for mothers. These findings may suggest closer links for fathers than for mothers between parental nurturing responses and psychological state, or they may reveal a stronger subjective bias for fathers inherent in the Q-sort method. PMID- 10791478 TI - Very preterm children who do not cooperate with assessments at three years of age: skill differences at five years. AB - This study examined skill differences at 5 years of age for very preterm children who were or were not cooperative with developmental testing at 3 years of age. All children born between 1986 and 1991 who were less than 30 weeks of gestation were followed prospectively. Two hundred one children were seen at both the 3- and 5-year assessments. Of the 201 children, 24 (11.9%) who had been uncooperative in the assessment at 3 years were seen at 5 years. Uncooperative children were matched to a group of cooperative children for sex, gestation, and/or birth weight. Nonparametric analyses revealed that scores on the Binet Pattern Analysis (p < .01) and the Bead Memory (p < .01) subtests were significantly different between the groups. The uncooperative children scored significantly more often in the at-risk range for tests of minor neurological dysfunction (MND; p < .01) compared with cooperative matched controls. The authors speculate that in very preterm children, uncooperative behavior shown at 3 years of age associated with poor visual/spatial skills and a high level of MND at 5 years of age may reveal children at risk for the development of nonverbal learning disabilities. PMID- 10791480 TI - Feeding interactions for children with cerebral palsy: contributions of mothers' psychological state and children's skills and abilities. AB - This study examined the relative importance of children's feeding skills and abilities and mothers' psychological state regarding their child's diagnostic status in predicting the observed quality of mother-child interaction during a naturalistic feeding situation. The study subjects were 69 children with cerebral palsy (CP), aged 14 to 53 months, and their mothers. Functional impairments of the children ranged from mild to severe. Mother-child interaction was observed during 20 minutes of feeding and was scored using global ratings of mothers' and children's socioemotional behavior. In addition, mothers' feeding interactions were dichotomously coded for sensitive and responsive behaviors. Children's feeding skills and abilities were assessed by maternal report and by observation. Mothers' resolution of grief associated with the child's diagnosis was assessed using standardized interview procedures. The findings strongly indicate that, contrary to expectations, the mothers' resolution status was not related to qualities of the feeding interactions (either dyadic or mothers' behavior) and that feeding interactions are best predicted for children with CP by their skills and abilities. The results are discussed in relation to the nature, determinants, and consequences of feeding in children with and without disabilities. PMID- 10791479 TI - Maternal child-rearing behaviors and correlates in rural minority areas of Yunnan, China. AB - An epidemiological survey of maternal child-rearing behaviors was conducted in the rural minority areas of Yunnan, China. An interview of 2019 rural mothers of Hani, Yi, Hui, Miao, and Han and 544 urban mothers with children younger than 7 years of age showed that 51.7% of these rural mothers placed their infants in swaddling clothes. In the rural sample, exclusive breastfeeding occurred at frequency rates of 76.7% and 34.2% during 3 and 6 months postpartum, respectively. Maternal training and playing behaviors were reported less frequently, whereas physical punishment for children's misbehavior was more commonly found in the child-rearing behavior of rural mothers. Maternal child rearing behaviors differed significantly among ethnic groups as well as between urban and rural areas. Maternal child-rearing behaviors were significantly associated with maternal education level, family type, family income, maternal age at delivery, and number of children in the family. Children's gender had no significant association with maternal child-rearing behaviors. PMID- 10791481 TI - Subspecialty certification in developmental-behavioral pediatrics: past and present challenges. PMID- 10791482 TI - Developmental-behavioral pediatrics: the American Board of Pediatrics' perspective. PMID- 10791483 TI - "What can I do to enhance the development of a premature infant with chronic lung disease?". AB - CASE. Timmy was born at 32 weeks of gestation after an uncomplicated pregnancy until there was a spontaneous rupture of the membranes and preterm labor associated with chorioamnionitis. A 2-month hospitalization in the neonatal intensive care unit (NICU) was associated with pneumonia, a Grade II intraventricular hemorrhage, chronic lung disease, and a slow weight gain in the nursery. He was discharged to home with plans for ongoing care by his pediatrician. The primary care pediatrician attended a multidisciplinary conference with the NICU staff and Timmy's parents. At the time of discharge from the nursery, at 38 weeks postconceptual age, Timmy still required oral diuretics and supplemental oxygen, as well as other medications such as iron. Timmy's respiratory rates were between 40 and 60 breaths per minute at rest, with mild intercostal retractions. He was discharged with a cardiorespiratory monitor. The discharge examination revealed mild to moderate symmetrical hypotonia with intact deep tendon reflexes, shoulder girdle weakness, and a mild head lag. Timmy would regard a human face and a bright object and would follow them briefly. He became active and would thrash his extremities with minimal tactile, bright light, or auditory stimulation. Typically, he settled slowly with swaddling and a pacifier. Nursing was slow to develop; he was currently receiving one half of his calories at the breast and the remainder of his calories from bottle-feeding of fortified expressed breast milk. As she prepared for the first office visit with Timmy and his parents, the pediatrician asked herself, "What can I do to enhance the developmental outcome for this child?" PMID- 10791484 TI - Methodological issues in analyzing psychological test scores in pediatric clinical trials. AB - Clinical trials to address drug dosing, safety, and efficacy issues in the pediatric population are becoming more common. In some studies, tests of mental ability are administered at regular intervals in drug trials for treatment of children with HIV, certain types of cancer, sickle cell anemia, and diabetes. The test scores are used to examine differences between treatments in efficacy and safety over time. In addition to the well-known problems of analyzing repeated measures with incomplete data profiles, the analyses of these data are complicated by a number of unique features, including that children can be so ill that their raw scores cannot be mapped to a normed scaled score, and that children may be in the studies long enough that they transition between the age appropriate instruments. These issues are often ignored in data analyses and can potentially cause incorrect conclusions regarding treatment efficacy and safety. This article describes these issues and their possible consequences. A simple approach to determine their impact on the statistical analysis of a particular clinical trial is suggested. The approach is illustrated with data from a Phase III trial in HIV-infected children. PMID- 10791485 TI - George L. Engel, M.D. December 10, 1913-November 26, 1999. PMID- 10791486 TI - A piece of my mind. Holding the bag. PMID- 10791488 TI - Delay in treating schizophrenia may narrow therapeutic window of opportunity. PMID- 10791487 TI - Antarctic medicine. PMID- 10791489 TI - Initiative seeks answers to cancer disparities. PMID- 10791490 TI - Consensus panel considers osteoporosis. PMID- 10791491 TI - From the Centers for Disease Control and Prevention. Community indicators of health-related quality of life--United States, 1993-1997. PMID- 10791492 TI - From the Centers for Disease Control and Prevention. Use of hospital discharge data to monitor uterine rupture--Massachusetts, 1990-1997. PMID- 10791493 TI - From the Centers for Disease Control and Prevention. Alternate two-dose hepatitis B vaccination schedule for adolescents aged 11-15 years. PMID- 10791494 TI - Access to health care for adolescents. PMID- 10791495 TI - Intra-arterial prourokinase for acute ischemic stroke. PMID- 10791496 TI - Intra-arterial prourokinase for acute ischemic stroke. PMID- 10791497 TI - Intra-arterial prourokinase for acute ischemic stroke. PMID- 10791498 TI - Preventing harm from thimerosal in vaccines. PMID- 10791499 TI - Preventing harm from thimerosal in vaccines. PMID- 10791500 TI - Mortality among elderly caregivers. PMID- 10791501 TI - Tobacco and alcohol advertisements in magazines: are young readers being targeted? PMID- 10791502 TI - A prospective study of diet quality and mortality in women. AB - CONTEXT: Most studies of diet and health care have focused on the role of single nutrients, foods, or food groups in disease prevention or promotion. Few studies have addressed the health effects of dietary patterns, which include complex mixtures of foods containing multiple nutrients and nonnutrients. OBJECTIVE: To examine the association of mortality with a multifactorial diet quality index. DESIGN AND SETTING: Data from phase 2 (1987-1989) of a prospective cohort study of breast cancer screening, the Breast Cancer Detection Demonstration Project, with a median follow-up of 5.6 years. PARTICIPANTS: A total of 42,254 women (mean age, 61.1 years) who completed the food frequency questionnaire portion of the survey. MAIN OUTCOME MEASURE: All-cause mortality by quartile of Recommended Food Score (RFS; the sum of the number of foods recommended by current dietary guidelines [fruits, vegetables, whole grains, low-fat dairy, and lean meats and poultry] that were reported on the questionnaire to be consumed at least once a week, for a maximum score of 23). RESULTS: There were 2065 deaths due to all causes in the cohort. The RFS was inversely associated with all-cause mortality. Compared with those in the lowest quartile, subjects in the upper quartiles of the RFS had relative risks for all-cause mortality of 0.82 (95% confidence interval [CI], 0.73-0.92) for quartile 2, 0.71 (95% CI, 0.62-0.81) for quartile 3, and 0.69 (95% CI, 0.61-0.78) for quartile 4 adjusted for education, ethnicity, age, body mass index, smoking status, alcohol use, level of physical activity, menopausal hormone use, and history of disease (chi2 for trend = 35.64, P<.001 for trend). CONCLUSIONS: These data suggest that a dietary pattern characterized by consumption of foods recommended in current dietary guidelines is associated with decreased risk of mortality in women. PMID- 10791503 TI - Problems with the interpretation of pharmacoeconomic analyses: a review of submissions to the Australian Pharmaceutical Benefits Scheme. AB - CONTEXT: Pharmacoeconomic analyses are being used increasingly as the basis for reimbursement of the costs of new drugs. Reports of these analyses are often published in peer-reviewed journals. However, the analyses are complex and difficult to evaluate. OBJECTIVE: To describe the nature of problems encountered in the evaluation and interpretation of pharmacoeconomic analyses used as a basis for reimbursement decisions. DATA SOURCES: All major submissions to the Department of Health and Aged Care (DHAC) by the pharmaceutical industry for funding made under the Australian Pharmaceutical Benefits Scheme. Specifically, the DHAC's database of submissions that were received between January 1994 and December 1997 were reviewed. STUDY SELECTION: Of a total of 326 submissions, 218 had serious problems of interpretation and were included in the analysis. The nature of the serious problems reviewed were classified as estimates of comparative clinical efficacy, comparator issues, modeling issues, and calculation errors. DATA EXTRACTION: All submissions in the DHAC's database were reviewed and data were extracted if both the DHAC evaluators and technical subcommittee considered problems to have a significant bearing on the decisions of the parent committee. DATA SYNTHESIS: Of a total of 326 submissions, 218 (67%) had significant problems and 31 had more than 1 problem. Of the 249 problems identified, 154 (62%) related to uncertainty in the estimates of comparative clinical efficacy, and 71 (28.5%) related to modeling issues, which included clinical assumptions or cost estimates, used in the construction of the economic models. There were 15 instances of disagreement over the choice of comparator, and serious calculation errors were found on 9 occasions. Overall, 159 problems (64%) were considered to be avoidable. CONCLUSIONS: Significant problems were identified in these pharmacoeconomic analyses. The intensive evaluation process used in the Australian Pharmaceutical Benefits Scheme allowed for identification and correction of pharmacoecomomic analysis problems, but the resources that are required may be beyond the capacity of many organizations, including peer reviewed journals. PMID- 10791504 TI - Impaired cerebral vasoreactivity and risk of stroke in patients with asymptomatic carotid artery stenosis. AB - CONTEXT: Standards for treating patients with asymptomatic carotid artery stenosis have been difficult to establish because of the lack of evidence for factors influencing these patients' prognoses. However, preliminary evidence suggests that an alteration in cerebral hemodynamic function may play a relevant role in the occurrence of stroke in patients with carotid artery disease. OBJECTIVE: To investigate the relationship between cerebrovascular reactivity to hypercapnia and cerebrovascular events in patients with severe unilateral asymptomatic carotid artery stenosis. DESIGN AND SETTING: Prospective, blinded longitudinal study conducted in an outpatient neurovascular department in Italy between June 1996 and April 1998, with a median follow-up of 28.5 months. PATIENTS: Ninety-four patients with asymptomatic carotid artery stenosis of at least 70% (74 men; mean age, 71 years). MAIN OUTCOME MEASURES: Subsequent occurrence of cerebral ischemic events (transient ischemic attack or stroke) or death, analyzed by cerebrovascular reactivity to hypercapnia (measured by transcranial Doppler ultrasonography and calculated by the breath-holding index values in the middle cerebral arteries). RESULTS: The overall annual rate for all ischemic events was 7.9%. Seventeen patients (18%) had ischemic events, all but 1 of which were ipsilateral to the carotid artery stenosis. Among factors considered, only lower breath-holding index values in the middle cerebral artery ipsilateral to carotid artery stenosis were significantly associated with the risk of an event (hazard ratio, 0.09; 95% confidence interval, 0.02-0.38; P=.001, by multivariate analysis). Based on data from previously studied healthy subjects, the cutoff of the breath-holding index for distinguishing between impaired and normal cerebrovascular reactivity was determined to be 0.69. Using this cutoff, the annual ipsilateral ischemic event risk was 4.1% in patients with normal and 13.9% in those with impaired breath-holding index values. CONCLUSIONS: These results suggest a link between impaired cerebrovascular reactivity and the risk of ischemic events ipsilateral to severe asymptomatic carotid stenosis. PMID- 10791505 TI - Acute onset of decreased vision and hearing traced to hemodialysis treatment with aged dialyzers. AB - CONTEXT: A recent event in which 7 patients at 1 hospital developed decreased vision and hearing, conjunctivitis, headache, and other severe neurologic symptoms 7 to 24 hours after hemodialysis drew attention to the issue of the long term integrity of dialysis machines and materials. OBJECTIVE: To determine the cause of the adverse reactions that occurred during this event. DESIGN, PATIENTS, AND SETTING: Retrospective cohort study of all 9 patients who received hemodialysis at hospital A on September 18, 1996, the day of the outbreak. A case patient was defined as any hospital A patient with acute onset of decreased vision and hearing and conjunctivitis after dialysis on that day. Non-case patients were all others who underwent dialysis at hospital A on that day but did not develop adverse reactions. In an attempt to reproduce the conditions of the event, cellulose acetate dialysis membranes of various ages were retrieved from other sources and tested for physical and chemical degradation, and degradation products were identified, characterized, and injected intravenously into rabbits. MAIN OUTCOME MEASURES: Clinical signs and symptoms, time to resolution of symptoms, mortality, and dialyzer type and age, for case- vs non-case-patients. RESULTS: Seven of the 9 patients met the case definition. In addition to diminished vision and hearing, conjunctivitis, and headache, some case-patients had blood leak alarm activation (n=6), confusion/lethargy (n=5), corneal opacification (n=4), cardiac arrest (n=2), or other neurologic signs and symptoms. One case-patient died during hospitalization after the event; 5 of 7 case-patients died within 13 months. Resolution of signs and symptoms varied but persisted more than 3 years or until death in 3 of the 6 patients who survived hospitalization. All case-patients but no non-case-patients were exposed to 11.5 year-old cellulose acetate dialyzers (all of these dialyzers were discarded by the hospital before our investigation). Laboratory investigation of field retrieved 0- to 13.6-year-old dialyzers of similar type indicated significant chemical degradation in the older membranes. In vivo injection of extracts of membrane degradation products produced iritis and hemorrhages in rabbits' eyes. CONCLUSIONS: Severe patient injury was associated with exposure to aged cellulose acetate membranes of dialyzers, allowing cellulose acetate degradation products to enter the blood. Clinicians should be aware that aged cellulose acetate membranes may cause severe adverse reactions. PMID- 10791506 TI - Factors associated with caregivers' choice of infant sleep position, 1994-1998: the National Infant Sleep Position Study. AB - CONTEXT: The success and simplicity of the 1994 national "Back to Sleep" campaign to reduce sudden infant death syndrome provides an opportunity to study which elements determine whether a behavior will change in the desired direction in response to a public health intervention. OBJECTIVE: To examine sociodemographic characteristics, motivation, and message exposure to ascertain which factors influenced a caregiver's choice of infant sleep position after implementation of the campaign. DESIGN: Annual nationally representative telephone surveys conducted between 1994 and 1998. SETTING: The 48 contiguous United States. PARTICIPANTS: Nighttime caregivers of infants born within the 7 months prior to interview between 1994 and 1998. Approximately 1000 interviews were conducted each year. MAIN OUTCOME MEASURES: The position the infant was usually placed in for sleep, sleep position recommendations received from specific sources, and reasons reported for position choice. RESULTS: Between 1994 and 1998, prone placement declined from 44% to 17% among white infants and from 53% to 32% among black infants. Supine placement increased from 27% to 58% among white infants and from 17% to 31% among black infants. During this period, reports of supine recommendations from at least 1 source doubled from 38% to 79%. From 1995 to 1998, 86% of caregivers who placed the infant prone reported receiving only nonprone recommendations. Infant comfort was given as a reason for prone placement by 82% of these caregivers. In multivariate analysis, physician recommendation of "supine not prone" had the strongest influence and was associated with decreased prone placement (odds ratio [OR], 0.25 [95% confidence interval [CI], 0.16-0.39]) and increased supine placement (OR, 3.37 [95% CI, 2.38 4.76]). Recommendations from all 4 sources (the physician, neonatal nurse, reading materials, and radio/television) further increased the probability of supine placement (OR, 6.01 [95% CI, 4.57-7.90]). Other factors independently associated with increased prone and decreased supine placement included maternal black race, parity of more than 1, and living in a southern or mid-Atlantic state. CONCLUSIONS: According to our study, as of 1998, approximately one fifth of infants were still placed prone, and only half were placed supine. Recommendations of supine placement during infancy by physicians at well-baby checks and by neonatal nursery staff and print and broadcast media have increased the proportion of infants placed supine. Caregiver beliefs regarding perceived advantages of prone sleeping should be addressed to attain further reduction in prone placement. PMID- 10791507 TI - A 48-year-old man with recurrent sinusitis. PMID- 10791508 TI - A 69-year-old man with anger and angina, 1 year later. PMID- 10791509 TI - Mortality among men using homeless shelters in Toronto, Ontario. AB - CONTEXT: Homeless persons in US cities have high mortality rates. However, few comparison data exist for death rates among homeless persons in other developed countries. OBJECTIVES: To compare mortality rates among men using homeless shelters and the general population in Toronto, Ontario, and to determine whether mortality rates differ significantly among men using homeless shelters in Canadian and US cities. DESIGN: Cohort study conducted from 1995 through 1997, with a mean follow-up of 2.6 years. PARTICIPANTS: Men aged 18 years or older who used homeless shelters in Toronto in 1995 (n=8933). MAIN OUTCOME MEASURE: Mortality rate ratios comparing age-specific mortality rates among men using homeless shelters in Toronto with those of men in the general population of Toronto and of men using homeless shelters in New York, NY; Boston, Mass; and Philadelphia, Pa. RESULTS: Men using homeless shelters in Toronto were more likely to die than men in the city's general population. Mortality rate ratios were 8.3 (95% confidence interval [CI], 4.4-15.6) for men aged 18 to 24 years, 3.7 (95% CI, 3.0-4.6) for men aged 25 to 44 years, and 2.3 (95% CI, 1.8-3.0) for men aged 45 to 64 years. In most cases, however, the risk of death was significantly lower for men using homeless shelters in Toronto than for those in US cities. For men aged 25 to 44 years using homeless shelters, mortality rate ratios were 0.52 (95% CI, 0.41-0.65) for Toronto compared with Boston and 0.61 (95% CI, 0.44-0.85) for Toronto compared with New York City. For men aged 35 to 54 years using homeless shelters, the mortality rate ratio was 0.42 (95% CI, 0.27 0.66) for Toronto compared with Philadelphia. CONCLUSIONS: Mortality rates among men who use homeless shelters in Toronto, while higher than in the general population of Toronto, are much lower than mortality rates observed among men using homeless shelters in 3 major US cities. Further study is needed to identify the reasons for this disparity. PMID- 10791510 TI - Pharmacoeconomic analyses: making them transparent, making them credible. PMID- 10791511 TI - Switching health plans to obtain drug coverage. PMID- 10791513 TI - JAMA Patient Page: healthy diet. PMID- 10791512 TI - Exhaustion of drug benefits and disenrollment of medicare beneficiaries from managed care organizations. AB - CONTEXT: Many Medicare beneficiaries enroll in managed care health plans to obtain outpatient drug benefits. Increasing pharmaceutical utilization and costs and decreasing drug benefits increase the likelihood that medication use by such enrollees will exceed drug benefits, which may lead to health plan disenrollment. OBJECTIVE: To test the hypothesis that exhaustion of managed care drug benefits by Medicare beneficiaries is associated with disenrollment from the health plan. DESIGN: Retrospective cohort study followed up for 1 year (1998) using an enrollment/claims database. SETTING: Four geographically diverse network-model health plans that had annual drug benefits of $300, $500, $600, or $1000. PARTICIPANTS: A total of 61,412 elderly Medicare beneficiaries. MAIN OUTCOME MEASURE: Voluntary disenrollment from health plans by members who did or did not exhaust their drug benefits. RESULTS: The likelihood of exhausting 1998 drug benefits ranged from 17% to 25% across health plans (P<.001). The relative hazards of disenrollment from the 4 plans when drug benefits had been exhausted were 2.5 (95% confidence interval [CI], 2.3-2.8), 1.9 (95% CI, 1.7-2.1), 2.7 (95% CI, 2.0-3.6), and 2.1 (95% CI, 1.9-2.4). Statistical adjustments for age, sex, prior enrollment, hospital admissions, physician visits, and county of residence did not alter these estimates. CONCLUSIONS: Exhaustion of drug benefits was associated with a significant increase in the likelihood of disenrollment of Medicare beneficiaries. This finding arouses concern that Medicare beneficiaries must change plans to have financial access to medications, which can lead to discontinuity in care and diversion of resources from care to administrative matters. Policymakers should strive to avoid fragmented systems of providing drug benefits. PMID- 10791514 TI - Measles, MMR, and autism: the confusion continues. PMID- 10791515 TI - Which surfactant for treatment of respiratory-distress syndrome. PMID- 10791516 TI - Who benefits from radiotherapy in treatment of endometrial cancer and at what price? PMID- 10791517 TI - Effect of hepatitis B virus mutants on efficacy of vaccination. PMID- 10791518 TI - Pulvinar sign on MRI images in variant Creutzfeldt-Jakob disease. PMID- 10791519 TI - Inhaled corticosteroids, bone density, and risk of fracture. PMID- 10791520 TI - Anatomical science in Vienna, 1938-45. PMID- 10791521 TI - Pumactant and poractant alfa for treatment of respiratory distress syndrome in neonates born at 25-29 weeks' gestation: a randomised trial. AB - BACKGROUND: Exogenous surfactant preparations vary in their constitution and biophysical properties. Synthetic and animal-derived preparations lower the rate of death compared with controls. No significant differences in mortality or important long-term clinical outcomes have been shown between them in randomised trials. We did a randomised controlled trial to compare pumactant, a synthetic surfactant, with poractant alfa, an animal-derived surfactant, both of which are widely used in the UK. METHODS: We enrolled 212 neonates born between 25 weeks' and 29 weeks and 6 days' gestation who were intubated for presumed surfactant deficiency and were free from life-threatening malformations. We randomly assigned 105 neonates poractant alfa, and 107 pumactant. The primary outcome was duration of high-dependency care and mortality was a secondary outcome. Analysis was by intention to treat. FINDINGS: Outcome data were analysed for 199 babies. The trial was stopped on the recommendation of the data and safety monitoring committee because mortality assumed a greater importance than the primary outcome. Predischarge mortality differed significantly between groups, in favour of poractant alfa (14.1 vs 31.0%, p=0.006; odds ratio 0.37 [95% CI 0.18-0.76). This difference was sustained after adjustment for centre, gestation, birthweight, sex, plurality, and use of antenatal steroids. INTERPRETATION: Mortality was unexpectedly lower among neonates who received poractant alfa than among those who received pumactant, and was independent of all the variables we investigated. Stopping the trial early may have widened the difference between the treatment groups. PMID- 10791522 TI - Effect of centre on outcome of bone-marrow transplantation for acute myeloid leukaemia. Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation. AB - BACKGROUND: There is increasing pressure for the recognition and replication of good clinical practice. We undertook a study to assess the variability in outcome of allogeneic bone-marrow transplantation among major European centres. METHODS: We studied 13 centres, including 522 patients (aged 16-55 years), which had undertaken more than 30 bone-marrow transplantations between Jan 1, 1987, and Dec 31, 1995, for acute myeloid leukaemia in first complete remission. We undertook a (global) multivariate analysis of all factors known previously to influence outcome and a stratified analysis that initially defined, by multivariate analysis, significant variables in this study and then used a proportional-hazard model including centres. FINDINGS: The overall results at 3 years were 57% (95% CI 53-61) for leukaemia-free survival (LFS), 23% (19-27) for relapse incidence (RI), and 26% (22-30) for treatment-related mortality (TRM) with a range for centres of 36-75%, 10-37%, and 8-54%, respectively. Both methods of analysis showed the centre effect to be highly significant for LFS and TRM, but not for RI. Variables associated with a significantly poor outcome were age over 43 years (p=0.01), time from diagnosis to first complete remission longer than 65 days (p=0.02), and centre (p=0.013) for LFS, and age over 43 years (p=0.023), time from first complete remission to transplantation of longer than 93 days (p=0.03), and centre (p=0.001) for TRM. Moreover, different centres had different prognostic criteria for good-risk or bad-risk patients indicating that risk factors do not have the same impact in each individual centre. INTERPRETATION: The outcome of bone-marrow transplantation for acute myeloid leukaemia in first complete remission is influenced by the centre in which the procedure is done, even with adjustment for known prognostic risk factors. Significant prognostic factors vary among centres, which means that the relative risk is not the same in each individual centre. However, centres may treat populations with different risks of as yet unidentified prognostic factors. Experience may partly account for the difference in outcome among centres. PMID- 10791523 TI - Inhaled corticosteroid use and bone-mineral density in patients with asthma. AB - BACKGROUND: Inhaled corticosteroids are absorbed into the systemic circulation, but the extent to which they have adverse effects on bone is uncertain. The question is important since 3% of the European population take an inhaled corticosteroid regularly and may do so for many years. METHODS: We studied the dose-response relation between cumulative inhaled corticosteroid dose and bone mineral density at the lumbar spine and proximal femur in 196 adults (119 women) with asthma aged 20-40 years. Patients had taken an inhaled corticosteroid regularly for at least 6 months, and had had limited exposure to systemic steroids. Cumulative dose of inhaled corticosteroid was calculated from questionnaires and computerised and written general-practice records, and its effect on bone-mineral density was estimated by multiple regression analysis. FINDINGS: Median duration of inhaled corticosteroid treatment was 6 years (range 0.5-24), and median cumulative dose was 876 mg (87-4380). There was a negative association between cumulative dose of inhaled corticosteroid and bone-mineral density at the lumbar spine (L2-L4), femoral neck, Ward's triangle, and trochanter, both before and after adjustment for the effects of age and sex. A doubling in dose of inhaled corticosteroid was associated with a decrease in bone mineral density at the lumbar spine of 0.16 SD (95% CI 0.04-0.28). Similar decreases were found at the femoral neck, Ward's triangle, and trochanter. Adjustment for potential confounding factors including physical activity and past oral, nasal, dermal, and parenteral corticosteroids did not weaken the associations. INTERPRETATION: This study provides evidence of a negative relation between total cumulative dose of inhaled corticosteroid and bone-mineral density in patients with asthma. PMID- 10791524 TI - Surgery and postoperative radiotherapy versus surgery alone for patients with stage-1 endometrial carcinoma: multicentre randomised trial. PORTEC Study Group. Post Operative Radiation Therapy in Endometrial Carcinoma. AB - BACKGROUND: Postoperative radiotherapy for International Federation of Gynaecology and Obstetrics (FIGO) stage-1 endometrial carcinoma is a subject of controversy due to the low relapse rate and the lack of data from randomised trials. We did a multicentre prospective randomised trial to find whether postoperative pelvic radiotherapy improves locoregional control and survival for patients with stage-1 endometrial carcinoma. METHODS: Patients with stage-1 endometrial carcinoma (grade 1 with deep [> or =50%] myometrial invasion, grade 2 with any invasion, or grade 3 with superficial [<50%] invasion) were enrolled. After total abdominal hysterectomy and bilateral salpingo-oophorectomy, without lymphadenectomy, 715 patients from 19 radiation oncology centres were randomised to pelvic radiotherapy (46 Gy) or no further treatment. The primary study endpoints were locoregional recurrence and death, with treatment-related morbidity and survival after relapse as secondary endpoints. FINDINGS: Analysis was done according to the intention-to-treat principle. Of the 715 patients, 714 could be evaluated. The median duration of follow-up was 52 months. 5-year actuarial locoregional recurrence rates were 4% in the radiotherapy group and 14% in the control group (p<0.001). Actuarial 5-year overall survival rates were similar in the two groups: 81% (radiotherapy) and 85% (controls), p=0.31. Endometrial-cancer-related death rates were 9% in the radiotherapy group and 6% in the control group (p=0.37). Treatment-related complications occurred in 25% of radiotherapy patients, and in 6% of the controls (p<0.0001). Two-thirds of the complications were grade 1. Grade 3-4 complications were seen in eight patients, of which seven were in the radiotherapy group (2%). 2-year survival after vaginal recurrence was 79%, in contrast to 21% after pelvic recurrence or distant metastases. Survival after relapse was significantly (p=0.02) better for patients in the control group. Multivariate analysis showed that for locoregional recurrence, radiotherapy and age below 60 years were significant favourable prognostic factors. INTERPRETATION: Postoperative radiotherapy in stage-1 endometrial carcinoma reduces locoregional recurrence but has no impact on overall survival. Radiotherapy increases treatment-related morbidity. Postoperative radiotherapy is not indicated in patients with stage-1 endometrial carcinoma below 60 years and patients with grade-2 tumours with superficial invasion. PMID- 10791525 TI - The pulvinar sign on magnetic resonance imaging in variant Creutzfeldt-Jakob disease. AB - BACKGROUND: There is a need for an accurate non-invasive diagnostic test for variant Creutzfeldt-Jakob disease (vCJD). We investigated the sensitivity and specificity of bilateral pulvinar high signal on magnetic resonance imaging (MRI) for the diagnosis of vCJD. METHODS: MRI from patients with vCJD and controls (patients with suspected CJD) were analysed. Scans were reviewed on two separate occasions by two neuroradiologists and scored for the distribution of changes, and likely final diagnosis. Scans from vCJD cases were reassessed to reach a consensus on all abnormalities. FINDINGS: We analysed 36 patients and 57 controls. vCJD patients were correctly identified based on bilateral pulvinar high signal in 29 of 36 and 32 of 36 cases on the first assessment by the two radiologists, and 32 of 36 and 31 of 36 on their second assessment. Bilateral increased pulvinar signal was identified in one of 57 and one of 57 controls on the first assessment and two of 57 and three of 57 controls on the second assessment. These reported changes in controls were graded as minimal/equivocal in six of seven patients and moderate in one (<0.5% of all control assessments). 80% of the assessments in vCJD cases were graded as moderate or substantial. On consensus review, 28 of 36 cases and none of 57 controls had prominent bilateral pulvinar signal-sensitivity 78% (95% CI 60-90%) and specificity 100% (95% CI 94 100%). Other common MRI features of vCJD were medial thalamic and periaqueductal grey matter high signal, and the notable absence of cerebral atrophy. Pulvinar high signal correlated with histological gliosis. INTERPRETATION: In the appropriate clinical context the MRI identification of bilaterally increased pulvinar signal is a useful non-invasive test for the diagnosis of vCJD. PMID- 10791526 TI - Efficacy of new, concise schedule for melarsoprol in treatment of sleeping sickness caused by Trypanosoma brucei gambiense: a randomised trial. AB - BACKGROUND: African trypanosomiasis is a fatal disease caused by protozoan parasites of the species Trypanosoma brucei. The disease has reached epidemic dimensions in various countries of central Africa. Treatment of the second stage is long and complicated, and is hampered by severe adverse reactions to the first line drug, melarsoprol. Despite these problems, melarsoprol is likely to remain the drug of choice for the next decade. We therefore did a randomised trial comparing the standard treatment schedule with a new, concise regimen. METHODS: The safety and efficacy of the new schedule were assessed in patients presenting to a hospital in Kwanza Norte, Angola with sleeping sickness. The control group followed the 26-day standard Angolan schedule of three series of four daily injections of melarsoprol at doses increasing from 1.2 to 3.6 mg/kg within each series, with a 7-day interval between series. The new treatment schedule comprised 10 daily injections of 2.2 mg/kg. Primary outcomes assessed were elimination of parasites, deaths attributed to treatment, and rate of encephalopathy. Analysis was by intention to treat. FINDINGS: Of 767 patients with second-stage disease, 500 were enrolled: 250 were assigned the standard schedule, and 250 the new schedule. 40 patients on the standard schedule and 47 on the new schedule had adverse events which resulted in treatment disruption or withdrawal. 50 patients on the standard regimen deviated or withdrew from treatment, compared with two on the new regimen. Parasitological cure 24 h after treatment was 100% in both groups; there were six deaths (all due to encephalopathy) 30 days after treatment in each group. The number of patients with encephalopathic syndromes was also the same in each group (14). Skin reactions were more common with the new treatment, but all could be resolved by additional medication or withdrawal of treatment. INTERPRETATION: Considering the economic and practical advantages of the new 10-day schedule over the standard 26 day treatment schedule, and the similarity of treatment outcome, the new schedule is a useful alternative to the present standard, especially in epidemic situations and in locations with limited resources. PMID- 10791527 TI - Cryptococcal meningitis, an endocrine emergency? PMID- 10791528 TI - Autoagglutination of malaria-infected red blood cells and malaria severity. AB - Red blood cells infected with Plasmodium falciparum can adhere to each other and so form large autoagglutinates. We show that this phenotype is common in field isolates and is strongly associated with severe malaria. PMID- 10791529 TI - Effect of local radiotherapy for bone pain on urinary markers of osteoclast activity. AB - Urinary markers of bone resorption, pyridinoline and deoxypyridinoline were measured before and at 4 weeks after radiotherapy for metastatic bone pain. An association was shown between relief of metastatic skeletal pain by radiotherapy and low marker concentrations before and after treatment, lending support to the hypothesis that relief of metastatic bone pain by radiotherapy relates to an effect on bone, rather than tumour physiology. PMID- 10791530 TI - Scotopic sensitivity in dyslexia and requirements for DHA supplementation. AB - Much interest is shown in reduced scotopic sensitivity in dyslexia and the possible role of docosahexaenoic-acid deficiency as a causative factor. However, we found that significant decreases in scotopic sensitivity are not a general characteristic of dyslexia, which may cast doubt on the value of DHA supplementation. PMID- 10791532 TI - Mobile workstation for anaesthesia and intensive-care medicine. AB - Our mobile monitoring and treatment station, for use in anaesthesia and intensive care, allows transport of patients around the medical unit without disconnection from medical equipment, with a maximum level of safety, and low workload and costs. PMID- 10791531 TI - Blood pressure in young adulthood and mortality from cardiovascular disease. AB - Blood pressure measured in young adult men is positively associated in later life with increased cardiovascular disease mortality. The implication of this finding is that risk of cardiovascular disease starts in early life. PMID- 10791533 TI - Report bodes ill for antioxidant supplementation. PMID- 10791534 TI - Salvage therapy for HIV-1 infection--the challenge grows. PMID- 10791535 TI - Chronic Lyme disease: the debate goes on. PMID- 10791536 TI - Women's rights, a tourist boom, and the power of khat in Yemen. PMID- 10791537 TI - Measles epidemic sweeps through Afghanistan. PMID- 10791538 TI - Epilepsy in elderly people. AB - The prevalence and incidence of epilepsy are highest in later life with around 25% of new cases occurring in elderly people, many of whom will have concomitant neurodegenerative, cerebrovascular, or neoplastic disease. Difficulties accepting the diagnosis are frequently compounded by its unpredictable nature. Those affected commonly lose confidence and independence. Seizures in older people can result in physical injury, adding to low morale. Complete control is achievable in around 70% of patients with antiepileptic drug treatment. Optimum management requires rapid investigation, accurate diagnosis, effective treatment, sympathetic education, and assured support. The emergence of seizure disorders in old age places an increasing burden on health-care facilities and costs. A coordinated programme among health-care workers is advised to maintain the independence and improve the quality of life of this vulnerable patient population. PMID- 10791539 TI - White cells 2: impact of understanding the molecular basis of haematological malignant disorders on clinical practice. AB - The molecular basis of many leukaemias is now known, allowing precise diagnosis. Treatment of chronic myeloid leukaemia is now possible by targeting of the BCR ABL tyrosine kinase. The underlying molecular abnormalities in acute leukaemias allow the outlook for individual patients to be assessed at diagnosis and therapy tailored accordingly. Analysis of V(H) genes in B-cell malignant disorders allows these to be placed in the hierarchy of B-cell development and may provide prognostically valuable information. PMID- 10791540 TI - Anatomical science at University of Vienna 1938-45. Senate Project of the University of Vienna. PMID- 10791541 TI - Adrenal response to glucocorticoid treatment. PMID- 10791542 TI - Adrenal response to glucocorticoid treatment. PMID- 10791543 TI - Adrenal response to glucocorticoid treatment. PMID- 10791544 TI - Adrenal response to glucocorticoid treatment. PMID- 10791545 TI - Prenylation inhibitors to treat proliferative renal disease. PMID- 10791546 TI - Nucleic acid amplification testing of hepatitis B virus. PMID- 10791547 TI - Dopamine-transporter density in patients with ADHD. PMID- 10791548 TI - Dopamine-transporter density in patients with ADHD. PMID- 10791549 TI - Human bornaviruses and laboratory strains. PMID- 10791550 TI - Human bornaviruses and laboratory strains. PMID- 10791551 TI - A marker for hepatocellular damage. PMID- 10791552 TI - Hepatitis B booster vaccination for healthcare workers. PMID- 10791553 TI - Surveillance of communicable diseases in the Balkans. Committee of the Balkan Network for Surveillance of Communicable Diseases. PMID- 10791554 TI - Killing Iraq's children. PMID- 10791555 TI - Syphilis increases in Manchester, UK. PMID- 10791556 TI - Publishing surgery. PMID- 10791557 TI - Sickle hemoglobin (HbS) allele and sickle cell disease: a HuGE review. AB - Sickle cell disease is caused by a variant of the beta-globin gene called sickle hemoglobin (Hb S). Inherited autosomal recessively, either two copies of Hb S or one copy of Hb S plus another beta-globin variant (such as Hb C) are required for disease expression. Hb S carriers are protected from malaria infection, and this protection probably led to the high frequency of Hb S in individuals of African and Mediterranean ancestry. Despite this advantage, individuals with sickle cell disease exhibit significant morbidity and mortality. Symptoms include chronic anemia, acute chest syndrome, stroke, splenic and renal dysfunction, pain crises, and susceptibility to bacterial infections. Pediatric mortality is primarily due to bacterial infection and stroke. In adults, specific causes of mortality are more varied, but individuals with more symptomatic disease may exhibit early mortality. Disease expression is variable and is modified by several factors, the most influential being genotype. Other factors include beta-globin cluster haplotypes, alpha-globin gene number, and fetal hemoglobin expression. In recent years, newborn screening, better medical care, parent education, and penicillin prophylaxis have successfully reduced morbidity and mortality due to Hb S. PMID- 10791558 TI - N-acetyltransferase polymorphisms and colorectal cancer: a HuGE review. AB - The two expressed genes coding for N-acetyltransferase (NAT) activity, NAT1 and NAT2, are located on chromosome 8 at 8p21.3-23.1 and are polymorphic. Both enzymes are capable of N-acetylation, O-acetylation, and N,O-acetylation and are implicated in the activation and detoxification of known carcinogens. Single base pair substitutions in NAT2 tend to occur in combination with other substitutions within the gene. As yet, less work has been done to characterize NAT1 allelic variants. Various methods for the detection of the reported polymorphisms exist. It is important to select a method that is appropriate to the population being studied. The functional significance of many NAT allelic variants has not been determined. Geographic and ethnic variation in the frequency of NAT2 genotypes associated with fast or intermediate acetylation has been observed. Insufficient data for NAT1 genotypes are available to reveal a clear geographic pattern. No consistent association has been found between acetylator phenotype or genotype and colorectal cancer. The lack of consistency can in part be accounted for by methodological factors, including limited statistical power. Possible interactions between the NAT genes and either environmental exposures or other polymorphic genes encoding xenobiotic metabolizing enzymes have been investigated in only a minority of these studies, and these studies have lacked statistical power to detect interactions. PMID- 10791560 TI - Occurrence of congenital heart defects in relation to maternal mulitivitamin use. AB - The purpose of this study was to assess the relation between maternal multivitamin use and risk for cardiac defects in the offspring, using a population-based approach. The Atlanta Birth Defects Case-Control study is a population-based case-control study of infants born between 1968 and 1980 to mothers residing in metropolitan Atlanta, Georgia. The 958 case infants with nonsyndromic cardiac defects were actively ascertained from multiple sources. The 3,029 infants without birth defects (control infants) were selected from birth certificates by stratified random sampling. Periconceptional multivitamin use, defined as reported regular use of multivitamins from 3 months before pregnancy through the first 3 months of pregnancy, was contrasted with no use during the same time period. Periconceptional multivitamin use was associated with a reduced risk for nonsyndromic cardiac defects in the offspring (odds ratio (OR) = 0.76; 95% confidence interval (Cl): 0.60, 0.97). The risk reduction was strongest for outflow tract defects (OR = 0.46; 95% Cl 0.24, 0.86) and ventricular septal defects (OR = 0.61; 95% Cl: 0.38, 0.99). No risk reduction was evident when multivitamin use was begun after the first month of pregnancy. If these associations are causal, the results suggest that approximately one in four major cardiac defects could be prevented by periconceptional multivitamin use. PMID- 10791559 TI - 5,10-Methylenetetrahydrofolate reductase gene variants and congenital anomalies: a HuGE review. AB - The enzyme 5,10-methylenetetrahydrofolate reductase (MTHFR) is involved in folate metabolism. The MTHFR gene is located on chromosome 1 (1p36.3), and two common alleles, the C677T (thermolabile) allele and the A1298C allele, have been described. The population frequency of C677T homozygosity ranges from 1% or less among Blacks from Africa and the United States to 20% or more among Italians and US Hispanics. C677T homozygosity in infants is associated with a moderately increased risk for spina bifida (pooled odds ratio = 1.8; 95% confidence interval: 1.4, 2.2). Maternal C677T homozygosity also appears to be a moderate risk factor (pooled odds ratio = 2.0; 95% confidence interval: 1.5, 2.8). The A 1298C allele combined with the C677T allele also could be associated with an increased risk for spina bifida. Some data suggest that the risk for spina bifida associated with C677T homozygosity may depend on nutritional status (e.g., blood folate levels, intake of vitamins) or on the genotype of other folate-related genes (e.g., cystathionine-beta-synthase and methionine synthase reductase). Studies of the C677T allele in relation to oral clefts, Down syndrome, and fetal anticonvulsant syndrome either have yielded conflicting results or have not been yet replicated. PMID- 10791561 TI - Relation of abdominal height to cardiovascular risk factors in young adults: the Bogalusa heart study. AB - Obesity and fat patterns are important predictors of coronary heart disease risk. The relations of abdominal height (sagittal diameter) and various obesity measures to coronary heart disease risk factors were examined in a community based sample of 409 Blacks and 1,011 Whites aged 20-38 years in Bogalusa, Louisiana (1995-1996). Obesity measures used included weight, waist circumference, waist:hip ratio, waist:height ratio, abdominal height, triceps and subscapular skinfold thicknesses, body mass index, and conicity index. Abdominal height was highly correlated with other obesity measures, especially waist circumference (0.937-0.944, p < 0.001), and was least correlated with height. In multivariate analysis, abdominal height was an independent predictor of levels of total cholesterol, triglycerides, very low density lipoprotein cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, glucose, and insulin and of systolic and diastolic blood pressures (p < 0.05 to p < 0.001), with total R2 values ranging from 0.13 to 0.52. Abdominal height contributed more to the prediction of blood pressure than did other measures of central obesity. In canonical analysis, abdominal height was correlated more strongly with the coronary disease risk factor variables as a group than were other obesity measures. These results suggest that abdominal height adds another dimension to measures of obesity in that it may help to assess a component of visceral fat that other measures miss. PMID- 10791562 TI - Prospective study of diet, lifestyle, and intermittent claudication in male smokers. AB - The association between dietary and lifestyle factors and intermittent claudication was investigated in the Finnish Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study. The cohort comprised 26,872 male smokers aged 50-69 years who were free of claudication at study entry. At baseline (1985-1988), subjects completed a diet history questionnaire. During a median follow-up period of 4 years (ending in spring 1993), 2,578 men reported symptoms of claudication on the Rose questionnaire, which was administered annually. Smoking status was assessed every 4 months. Smoking, systolic blood pressure, serum total cholesterol, and diabetes mellitus were positively associated with risk for claudication, whereas serum high density lipoprotein cholesterol, education, and leisure time exercise were inversely associated with risk. Dietary carbohydrates, fiber, and n-6 polyunsaturated fatty acids were inversely associated with risk for claudication, as were some dietary and serum antioxidants: dietary vitamin C (highest quartile vs. lowest: relative risk (RR) = 0.86; 95% confidence interval (CI): 0.77, 0.97), dietary gamma-tocopherol (RR = 0.89; 95% CI: 0.79, 1.00), dietary carotenoids (RR = 0.82; 95% CI: 0.73, 0.92), serum alpha-tocopherol (RR = 0.88; 95% CI: 0.77, 1.00), and serum beta-carotene (RR = 0.77; 95% CI: 0.68, 0.86). Smoking cessation reduced subsequent risk for claudication (RR = 0.86; 95% CI: 0.75, 0.99). The authors conclude that classical risk factors for atherosclerosis are associated with claudication. High intakes of antioxidant vitamins may be protective. Further research is needed before antioxidants can be recommended for the prevention of intermittent claudication. PMID- 10791563 TI - Occupational risk factors for lung cancer in the French electricity and gas industry: a case-control survey nested in a cohort of active employees. AB - The main aim of this study was the analysis of occupational lung cancer risk factors in the French national electricity and gas company (Electricite de France Gaz de France (EDF-GDF)). A case-control survey nested in a cohort of male employees was undertaken. The study population consisted of all male staff who were active at EDF-GDF between January 1, 1978, and December 31, 1989. During this period, 310 cases of lung cancer were identified in the cancer register set up by the medical department of the company. For each case, four age-matched controls who were free of cancer at the time of occurrence of the case's lung cancer were randomly selected. Occupational exposures to 21 chemical agents were assessed for each subject using a job exposure matrix. The associations between lung cancer and the different agents were estimated using conditional logistic regression analysis. After adjustment for various occupational confounding factors, the analysis showed increased lung cancer risks linked to exposure to crystalline silica (highest exposure class: odds ratio = 2.27; 95% confidence interval: 1.10, 4.68) and creosotes (highest exposure level: odds ratio = 2.14; 95% confidence interval: 1.06, 4.31), with significant dose-response relationships for both exposures. PMID- 10791565 TI - Editing data: what difference do consistency checks make? AB - In 1998, the Florida Department of Health undertook a self-administered school based survey of tobacco use, attitudes, and behaviors among nearly 23,000 public school students in grades 6-12. The survey design did not use skip patterns; therefore, students had multiple opportunities to contradict themselves. By using examples from the high school portion (grades 9-12) of the survey, the authors examined five possible approaches to handling data inconsistencies and the effect that each has on point estimates. Use of these approaches resulted in point estimates of current cigarette use ranging from 25.6% to 29.7%. The number of missing respondents varied from 33 (less than 1%) to 1,374 (13%), depending on which approach was used. After stratification by gender and race, the prevalence estimates changed marginally for girls but strikingly for boys. Non-Hispanic White students were substantially more likely than non-Hispanic Black students to report current cigarette use, but the magnitude of this difference varied significantly according to the analytical approach used. The approach used to check data consistency may influence point estimates and comparability with other studies. Therefore, this issue should be addressed when findings are reported. PMID- 10791566 TI - Bias associated with study protocols in epidemiologic studies of disease familial aggregation. AB - The effect of selection bias has not been well evaluated in epidemiologic studies which focus on familial aggregation. The authors illustrate this type of bias for a reconstructed cohort study. With the reconstructed cohort design, cases and controls are first selected from the population and their relatives form the exposed and unexposed cohorts, respectively. The recurrence risk ratio (RRR) is calculated to assess and measure familial aggregation. The ways of utilizing information from relatives affects the estimate of RRR, and the authors show that a traditional method used in epidemiologic studies can yield a severely biased estimate of the RRR. However, this traditional approach can give approximately unbiased estimates under special conditions. A novel selection approach is proposed which yields an unbiased estimate of RRR. In conclusion, when relatives are identified through cases or controls, they should be included and counted in the study cohorts each time a case or control is selected, even if they or other family members have already been included. PMID- 10791564 TI - Trace element levels in drinking water and cognitive function among elderly Chinese. AB - The relation between trace element levels in drinking water and cognitive function was investigated in a population-based study of elderly residents (n = 1,016) in rural China in 1996-1997. Cognitive function was measured using a Chinese translation of the Community Screening Interview for Dementia. A mixed effects model was used to evaluate the effect of each of the elements on cognitive function while adjusting for age, sex, and educational level. Several of the elements examined had a significant effect on cognitive function when they were assessed in a univariate context. However, after adjustment for other elements, many of these results were not significant. There was a significant quadratic effect for calcium and a significant zinc-cadmium interaction. Cognitive function increased with calcium level up to a certain point and then decreased as calcium continued to increase. Zinc showed a positive relation with cognitive function at low cadmium levels but a negative relation at high levels. PMID- 10791567 TI - Re: "Geographic variation in sarcoidosis in South Carolina: its relation to socioeconomic status and health care indicators". PMID- 10791568 TI - Adrenergic regulation of Rana balcanica erythrocyte pyruvate kinase. AB - The regulation of pyruvate kinase activity by noradrenaline was investigated in Rana balcanica red cells. Thirty minutes of noradrenaline incubation induced a significant increase in the Vo/Vmax ratio of pyruvate kinase. The S0.5 for phosphoenolpyruvate of the enzyme significantly increased in the presence of noradrenaline while the Km for ADP decreased. In response to hormonal stimulation the Na +/H+ exchange was activated as was shown by the increase in Na+ and cyclic adenosine monophosphate from the 3rd min of incubation. All these effects were specific to alpha1 and beta antagonists. High concentrations of fructose diphosphate significantly activated the enzyme in the presence of noradrenaline but not in its absence. Furthermore, the presence of noradrenaline partially released the inhibition of the enzyme by adenosine triphosphate, inorganic phosphate and 2,3-diphosphoglycerate. The results suggest that noradrenaline stimulates glycolysis through pyruvate kinase activation. The mechanism of stimulation may is through Na+/H+ exchange activation, cyclic adenosine monophosphate concentration and Na(+)-K(+)-ATPase activation. PMID- 10791569 TI - Resistance to freshwater exposure in White Sea Littorina spp. I: Anaerobic metabolism and energetics. AB - Anaerobic metabolism and changes in the osmotic concentration of extravisceral fluid were studied in the White Sea periwinkles (Littorina littorea, Littorina saxatilis and Littorina obtusata) during freshwater exposure. Resistance to hypoosmotic stress increased in the order: L. obtusata < L. saxatilis < L. littorea. Our data suggest that osmotic shock is not a primary reason for mortality of the periwinkles under these conditions. During environmental anaerobiosis, considerable succinate accumulation (up to 10(-19) micromol g(-1) wet weight), and depletion of phosphagen and ATP pools were found in the studied species. Other metabolic end products (alanopine, strombine, lactate, acetate or propionate) were not detected. Succinate accumulation and net ATP breakdown were the fastest in the least resistant species, L. obtusata, and slowest in the most resistant, L. littorea. Rate of ATP turnover decreased during freshwater exposure in L. littorea and L. saxatilis, but not in L. obtusata. Our data suggest that differential resistance of three studied Littorina spp. to extreme hypoosmotic stress may be related to their different abilities to reduce metabolic rate and ATP turnover during sustained anoxia. Species-specific variations in anaerobic capacity of Littorina spp. are discussed in relation to their vertical distribution, size and ecology. PMID- 10791570 TI - Resistance to freshwater exposure in White Sea Littorina spp. II: Acid-base regulation. AB - Parameters of acid-base and energy status were studied by in vivo 31P-nuclear magnetic resonance spectroscopy in three White Sea Littorina spp. (L. littorea, L. saxatilis and L. obtusata) during prolonged anaerobiosis in freshwater. Intracellular pH decreased significantly, especially during the early period of anaerobiosis, but later the decrease in intracellular pH slowed down considerably, suggesting a capacity for intracellular pH regulation in all three species. There was a trend for intracellular pH to fall most rapidly in the least freshwater-resistant species, L. obtusata, as compared to the most resistant, L. littorea. Non-bicarbonate, non-phosphate buffer values estimated by the homogenate technique were similar in the three studied species (28-37 mmol pH(-1) kg(-1) wet weight) and did not change during freshwater exposure. The CaCO3 buffer value of the foot tissues was considerably higher (171-218 mmol pH(-1) kg( 1) wet weight) and decreased significantly during freshwater exposure. The contribution of the multiple tissue buffering systems to intracellular pH regulation in Littorina spp. shifts between different stages of freshwater exposure. Initially, the non-bicarbonate, non-phosphate tissue buffering system seems to be of major importance for metabolic proton buffering at intracellular pH between 7.5 and 7.0. During later stages of anaerobiosis and at lower intracellular pH, the CaCO3 buffer is involved in proton buffering. Decrease in the CaCO3 buffer value during freshwater exposure was in quantitative agreement with the amount of metabolic protons buffered, thus suggesting that CaCO3 tissue stores may serve as a major buffering system during prolonged anaerobiosis in Littorina spp. PMID- 10791571 TI - Thermal acclimation of locomotor performance in tadpoles and adults of the aquatic frog Xenopus laevis. AB - Among amphibians, the ability to compensate for the effects of temperature on the locomotor system by thermal acclimation has only been reported in larvae of a single species of anuran. All other analyses have examined predominantly terrestrial adult life stages of amphibians and found no evidence of thermal acclimatory capacity. We examined the ability of both tadpoles and adults of the fully aquatic amphibian Xenopus laevis to acclimate their locomotor system to different temperatures. Tadpoles were acclimated to either 12 degrees C or 30 degrees C for 4 weeks and their burst swimming performance was assessed at four temperatures between 5 degrees C and 30 degrees C. Adult X. laevis were acclimated to either 10 degrees C or 25 degrees C for 6 weeks and their burst swimming performance and isolated muscle performance was determined at six temperatures between 5 degrees C and 30 degrees C. Maximum swimming performance of cold-acclimated X. laevis tadpoles was greater at cool temperatures and lower at the highest temperature in comparison with the warm-acclimated animals. At the test temperature of 12 degrees C, maximum swimming velocity of tadpoles acclimated to 12 degrees C was 38% higher than the 30 degrees C-acclimation group, while at 30 degrees C, maximum swimming velocity of the 30 degrees C acclimation group was 41% faster than the 12 degrees C-acclimation group. Maximum swimming performance of adult X. laevis acclimated to 10 degrees C was also higher at the lower temperatures than the 25 degrees C acclimated animals, but there was no difference between the treatment groups at higher temperatures. When tested at 10 degrees C, maximum swimming velocity of the 10 degrees C-acclimation group was 67% faster than the 25 degrees C group. Isolated gastrocnemius muscle fibres from adult X. laevis acclimated to 10 degrees C produced higher relative tetanic tensions and decreased relaxation times at 10 degrees C in comparison with animals acclimated to 25 degrees C. This is only the second species of amphibian, and the first adult life stage, reported to have the capacity to thermally acclimate locomotor performance. PMID- 10791572 TI - Characterization of ATP-dependent proteolysis in embryos of the brine shrimp, Artemia franciscana. AB - Under anoxia, embryos of Artemia franciscana enter a state of quiescence. During this time protein synthesis is depressed, and continued degradation of proteins could jeopardize the ability to recover from quiescence upon return to favorable conditions. In this study, we developed an assay for monitoring ATP/ ubiquitin dependent proteolysis in order to establish the presence of this degradation mechanism in A. franciscana embryos, and to describe some characteristics that may regulate its function during anoxia-induced quiescence. For lysates experimentally depleted of adenylates, supplementation with ATP and ubiquitin stimulated protein degradation rates by 92 +/- 17% (mean +/- SE) compared to control rates. The stimulation by ATP was maximal at concentrations > or =11 micromol x l(-1). In the presence of ATP and ubiquitin, ubiquitin-conjugated proteins were produced by lysates during the course of the 4-h assays, as detected by Western blotting. Acute acidification of lysates to values approximating the intracellular pH observed under anoxia completely inhibited ATP/ubiquitin-dependent proteolysis. Depressed degradation was also observed under conditions where net ATP hydrolysis occurred. These results suggest that ATP/ubiquitin-dependent proteolysis is markedly inhibited under cellular conditions promoted by anoxia. Inhibition of proteolysis during quiescence may be one critical factor that increases macromolecular stability, which may ultimately govern the duration of embryo survival under anoxia. PMID- 10791573 TI - RNA turnover and protein synthesis in fish cells. AB - Protein synthesis in fish has been previously correlated with RNA content. The present study investigates whether protein and RNA synthesis rates are similarly related. Protein and RNA synthesis rates were determined from 3H-phenylalanine and 3H-uridine incorporation, respectively, and expressed as % x day(-1) and half lives, respectively. Three fibroblast cell lines were used: BF-2, RTP, CHSE 214, which are derived from the bluegill, rainbow trout and Chinook salmon, respectively. These cells contained similar RNA concentrations (approximately 175 microg RNA x mg(-1) cell protein). Therefore differences in protein synthesis rates, BF-2 (31.3 +/- 1.8)>RTP (25.1 +/- 1.7)>CHSE 214 (17.6 +/-1.1), were attributable to RNA translational efficiency. The most translationally efficient RNA (BF-2 cells), 1.8 mg protein synthesised x microg(-1) RNA x day(-1), corresponded to the lowest RNA half-life, 75.4 +/- 6.4 h. Translationally efficient RNA was also energetically efficient with BF-2 cells exploiting the least costly route of nucleotide supply (i.e. exogenous salvage) 3.5-6.0 times more than the least translationally efficient RNA (CHSE 214 cells). These data suggest that differential nucleotide supply, between intracellular synthesis and exogenous salvage, constitutes the area of pre-translational flexibility exploited to maintain RNA synthesis as a fixed energetic cost component of protein synthesis. PMID- 10791575 TI - Torpor, thermal biology, and energetics in Australian long-eared bats (Nyctophilus). AB - Previous studies have suggested that Australian long-eared bats (Nyctophilus) differ from northern-hemisphere bats with respect to their thermal physiology and patterns of torpor. To determine whether this is a general trait of Australian bats, we characterised the temporal organisation of torpor and quantified metabolic rates and body temperatures of normothermic and torpid Australian bats (Nyctophilus geoffroyi, 7 g and N. gouldi, 10 g) over a range of air temperatures and in different seasons. The basal metabolic rate of normothermic bats was 1.36 +/- 0.17 ml g(-1) h(-1) (N. geoffroyi) and 1.22 +/- 0.13 ml g(-1) h(-1) (N. gouldi), about 65% of that predicted by allometric equations, and the corresponding body temperature was about 36 degrees C. Below an air temperature of about 25 degrees C bats usually remained normothermic for only brief periods and typically entered torpor. Arousal from torpor usually occurred shortly after the beginning of the dark phase and torpor re-entry occurred almost always during the dark phase after normothermic periods of only 111 +/- 48 min (N. geoffroyi) and 115 +/- 66 min (N. gouldi). At air temperatures below 10 degrees C, bats remained torpid for more than 1 day. Bats that were measured overnight had steady state torpor metabolic rates representing only 2.7% (N. geoffroyi) and 4.2% (N. gouldi) of the basal metabolic rate, and their body temperatures fell to minima of 1.4 and 2.3 degrees C, respectively. In contrast, bats measured entirely during the day, as in previous studies, had torpor metabolic rates that were up to ten times higher than those measured overnight. The steady-state torpor metabolic rate of thermoconforming torpid bats showed an exponential relationship with body temperature (r2 = 0.94), suggesting that temperature effects are important for reduction of metabolic rate below basal levels. However, the 75% reduction of metabolic rate between basal metabolic rate and torpor metabolic rate at a body temperature of 29.3 degrees C suggests that metabolic inhibition also plays an important role. Torpor metabolic rate showed little or no seasonal change. Our study suggests that Australian Nyctophilus bats have a low basal metabolic rate and that their patterns of torpor are similar to those measured in bats from the northern hemisphere. The low basal metabolic rate and the high proclivity of these bats for using torpor suggest that they are constrained by limited energy availability and that heterothermy plays a key role in their natural biology. PMID- 10791574 TI - Gastrointestinal blood flow in the red Irish lord, Hemilepidotus hemilepidotus: long-term effects of feeding and adrenergic control. AB - Cardiac output, blood flow to the coeliac and mesenteric arteries, dorsal aortic blood pressure and heart rate were recorded simultaneously at rest and postprandial for 6 days in a teleost, the red Irish lord (Hemilepidotus hemilepidotus). We anticipated that gastrointestinal blood flow would increase postprandially, supported by an increase in cardiac output. However, we had no predictions for either the exact time-course of this response, or for the regional distribution of blood flow between to the two major arteries comprising the splanchnic circulation. In resting, unfed animals, blood flow to the coeliac artery and mesenteric artery was 4.1 +/- 0.6 ml min(-1) kg(-1) and 4.9 +/-1.3 ml min(-1) kg(-1), respectively (mean +/- SEM, n = 7), which together represented 34% of cardiac output. Feeding increased blood flow to the coeliac and mesenteric arteries in a time-dependent manner. The increase in coeliac artery blood flow preceded that in the mesenteric artery, a finding that is consistent with the coeliac artery supplying blood to the liver and stomach, while the mesenteric artery supplies blood to the stomach and intestine. Coeliac blood flow had increased by 84 +/- 18% after 1 day and had a peak increase of 112 +/- 40% at day 4 postprandial. Mesenteric blood flow was not significantly elevated at day 1, but had increased by 94 +/- 19% at day 4 postprandial. Cardiac output also increased progressively, increasing by a maximum of 90 +/- 30% at day 4. Because the increase in cardiac output was adequate to meet the postprandial increase in gut blood flow, the postprandial decreases in vascular resistance for the coeliac and mesenteric circulations mirrored the increases in blood flow. Intra-arterial injections of adrenaline and noradrenaline into resting fish more than doubled coeliac and mesenteric vascular resistances, and blood flow decreased proportionately. This adrenergic vasoconstriction was totally abolished by pretreatment with the alpha-adrenoceptor antagonist phentolamine, which in itself approximately halved coeliac and mesenteric vascular resistances. These observations indicate a significant alpha-adrenergic tone in the gastrointestinal circulation of the red Irish lord, the loss of which could not entirely account for the post-prandial increase in gastrointestinal blood flow. Other control mechanisms are suggested. PMID- 10791576 TI - Excretion of corticosteroids in urine and faeces of hares (Lepus europaeus). AB - Increased production of glucocorticoids by the adrenal cortex is found in mammals under stress. As cortisol itself is absent in the faeces, an enzyme immunoassay (11-oxoaetiocholanolone) measuring 11,17-dioxoandrostanes has already been established to measure faecal cortisol metabolites in ruminants for non-invasive monitoring of adrenocortical activity. The aim of this study was to establish route and delay of excretion of glucocorticoids in hares and to determine whether a cortisol-, corticosterone- or this new enzyme immunoassay is best suited to detect faecal glucocorticoid metabolites. In the first experiment radioactive labelled glucocorticoids (14C-cortisol and 3H-corticosterone) were administered intravenously to two groups of three hares in metabolic cages. All voided urine and faecal samples were collected for 4 days. Metabolites of both steroids were found predominantly in the urine (91 +/- 4%). Peak concentrations were observed in the first urinary sample following infusion (13 +/- 6 h) and in the faeces with a delay of about 1 day (23 +/- 7 h). Most of the radioactivity was not extractable with diethylether, indicating that the metabolites excreted in urine and faeces are mainly conjugated or polar unconjugated ones. This was confirmed by reverse-phase high-performance liquid chromatography separations of the metabolites, which also revealed marked differences concerning the metabolism of the two glucocorticoids injected. Compared with the cortisol and the corticosterone enzyme immunoassay, only the group-specific enzyme immunoassay for 11,17-dioxoandrostanes detected high quantities of immunoreactive metabolites. In a second experiment hares (n = 20) were stressed by rousing them three times (5 min, 10 min and another 5 min) with a 20-min break in-between. Faecal samples were collected 2 days before until 4 days after stress and analysed using the 11 oxoaetiocholanolone enzyme immunoassay. After stress significantly (P < 0.001) increased 11,17-dioxoandrostane concentrations were found. Based on these results, measuring 11,17-dioxoandrostanes in faeces enables non-invasive monitoring of disturbances in hares and thus provides a tool for field investigations elucidating the role of stress in hare populations. PMID- 10791578 TI - Radiologic case study. Post-transfusional reticuloendothelial system iron overload of sickle cell disease, secondary hemochromatosis. PMID- 10791577 TI - Modulation of lipogenic enzymes, fatty acid synthase and delta9-desaturase, in relation to migration in the western sandpiper (Calidris mauri). AB - Long-distance migration in birds is characterized physiologically by periods of rapid fattening and lipogenesis, and increased desaturation of fatty acids stored in adipose tissue. We investigated seasonal, age- and sex-related differences in activities of two lipogenic enzymes, fatty acid synthase and delta9-desaturase, in relation to migration in the small, Arctic-nesting western sandpiper (Calidris mauri). Migration, and associated lipogenesis and fattening, involved marked upregulation of these enzymes in this species. However, this increase in enzyme activity was only seen in actively migrating birds during spring migration, when fatty acid synthase and delta9-desaturase levels increased by 53% and 113%, respectively, compared to non-migrating birds. There was no change in fatty acid synthase enzyme activity during the premigration period, even though body mass of adult birds increased significantly during this period. Similarly, there was no increase in delta9-desaturase activity during premigration, despite the fact that birds increase the proportion of monounsaturated fatty acids in their fat stores at this time. We suggest that upregulation of lipogenic enzymes is required to support high rates of mass gain (0.4 g day(-1)) during short (1-4 day) periods at stop-over sites. However, slower rates of mass gain (0.09 g day(-1)) over several weeks prior to migration can be achieved without any increase in tissue-specific enzyme activity. PMID- 10791579 TI - Listen to your doctor. PMID- 10791580 TI - Posterior stabilization of unstable sacroiliac injuries with the Texas Scottish Rite Hospital spinal instrumentation. AB - This article describes a new alternative technique for stabilization of unstable sacroiliac injuries using the Texas Scottish Rite Hospital (TSRH) instrumentation. The TSRH instrumentation was applied in 14 consecutive patients with unstable posterior pelvic ring disruptions. All patients underwent follow-up for an average of 29 months. Clinical results, using the rating system of d'Aubigne, were good in 9 and satisfactory in 5 patients. No pressure sores, infection, significant loss of correction, or hardware failures were observed postoperatively. The use of the TSRH instrumentation for stabilization of sacroiliac injuries offered sufficient stabilization of the posterior pelvic ring and permitted early mobilization without loss of correction. PMID- 10791581 TI - Primary lesions of the patella. PMID- 10791582 TI - Rotator cuff tears and associated nerve injuries. AB - A series of 15 patients with concomitant rotator cuff tears and infraclavicular brachial plexus injuries treated between 1980 and 1989 were reviewed. There were 6 men and 9 women with a mean age of 65 years. Seventeen nerve injuries were identified, including 12 axillary nerves, 4 suprascapular nerves, and 1 musculocutaneous nerve. One patient had an injury to all three nerves. Thirteen patients underwent operative repair of the torn rotator cuff, and 2 patients who refused surgery were treated conservatively. The average time from injury to surgery was 7.7 months. Follow-up averaged 5.5 years (range: 2-10 years). Clinical results were graded according to pain, range of motion, and strength. Postoperatively, mean active forward elevation was 137 degrees and mean active external rotation was 40 degrees. Clinically, 8 patients achieved complete nerve recovery and 7 had an incomplete recovery. Satisfactory pain relief was achieved in 87% of patients with 60% having excellent or good function. Overall, the results of rotator cuff repair with concurrent nerve injury are less favorable than those of isolated cuff repairs. Careful preoperative assessment of concomitant nerve injury should be performed to better predict outcome. PMID- 10791583 TI - Cost effectiveness of outpatient anticoagulant prophylaxis after total hip arthroplasty. AB - Guidelines for deep venous thrombosis (DVT) and pulmonary embolism (PE) prophylaxis have been developed for patients undergoing total hip arthroplasty (THA). Studies suggest that risk for developing these complications may exist for as long as 3 months following surgery. Cost-effectiveness analyses were performed on three pharmacoprophylaxis regimens administered over a 30-day period using literature-reported values for incidences of DVT and PE in patients postdischarge following THA. A cost savings of $21,466.89 will occur for each thromboembolic event avoided if low-dose warfarin daily is used routinely compared to enoxaparin 40 mg daily. Additionally, a cost savings of $18,618.10 is experienced if enoxaparin 40 mg daily for 4 days plus low-dose warfarin daily is administered versus enoxaparin 40 mg daily. Clinicians may choose to continue prophylaxis postdischarge with enoxaparin 40 mg daily for 4 days in combination with warfarin for 30 days in these patients until results of more definitive studies become available. PMID- 10791584 TI - The use of elastic nails for intramedullary fixation of humeral fractures and nonunions. AB - From May 1993 to January 1997, a total of 130 humeral fractures and nonunions were treated using an "elastic" unreamed nail. The elastic nail consists of a cylindrical proximal component tapered for the application of the impactor extractor, distally containing the proximal ends of four or five secondary nails preloaded to diverge and held together by a retaining system. The surgical approach is through the olecranic fossa apex. Once introduced, the proximal end of the nail locks automatically by diverging the secondary nails in the proximal humeral epiphysis. Distally, the elastic nail is locked with a crossbolt. The elastic nail enables stable fixation of fractures or nonunions and allows early rehabilitation. PMID- 10791585 TI - Prevalence, clinical features, and risk factors of osteonecrosis of the femoral head among adults with sickle cell disease. AB - This prospective study examined the prevalence, clinical features, and risk factors of osteonecrosis of the femoral head among adult sickle cell disease patients in Guadeloupe. Screening of osteonecrosis of the femoral head was performed using radiography, bone scintigraphy, and tomodensitometry. One hundred thirteen adults with sickle cell disease (67 SS and 46 SC patients) comprised the study population. Forty-two (37.2%) patients had osteonecrosis of one or both hips (67 [29.6%] hips) without association to a particular genotype, although bilateral involvement was more frequent among SS patients. While the prevalence of femoral head osteonecrosis increased with age, patients of all ages were affected, particularly young SC adults. Osteonecrosis of the femoral head was diagnosed at preradiographic stages (stage I) in 30% of hips and was frequently asymptomatic (60% of all cases; 95% and 90% of stages I and II, respectively). Osteonecrosis of the femoral head was significantly associated with a history of leg ulcer and osteonecrosis of the humeral head. SS patients with higher hemoglobin levels had an increased risk of osteonecrosis of the femoral head. PMID- 10791586 TI - Magnetic resonance imaging evaluation and arthroscopic resection of localized pigmented villonodular synovitis of the knee. AB - Diagnosis of localized pigmented villonodular synovitis is clinically difficult, and plain radiographs are usually normal. This article presents five patients with localized pigmented villonodular synovitis of the knee. Symptoms suggested a meniscal lesion in three patients and a loose body in two. Magnetic resonance imaging performed prior to arthroscopic resection revealed a preoperative diagnosis of a tumor in all five patients. Magnetic resonance imaging is a valuable clinical tool for the assessment of intra-articular soft-tissue tumors of the knee that may otherwise be misdiagnosed. PMID- 10791587 TI - Computed tomography in the determination of transarticular C1-C2 screw length. AB - This study evaluated the significance of computed tomographic (CT) measurements of the upper cervical vertebrae and their clinical implications in transarticular C1-C2 screw placement. In the first part of the study, analysis of axial CT scans of the atlas of 46 patients who had a normal C1-C2 region was performed. Measurements included the vertical distance between the middle of the ventral cortex of the lateral mass and the anterior-most point of the anterior tubercle, and the angle of the anterior ring of C1 relative to the frontal plane. In the second part, axial CT scans of the upper cervical spine were performed in seven cadaveric cervical spines and analyzed using the same criteria. Using the Magerl technique of transarticular C1-C2 screw placement, one screw was placed in each cervical spine. Following each placement, a strict lateral radiograph was taken and the distance between the tip of the screw and the anterior-most point of the anterior tubercle of C1 was measured. Analysis of the cervical cadaveric specimens showed the vertical distance between the middle of the ventral cortex and the anterior-most part of the anterior tubercle when measured on CT scan corresponded to the distance measured on lateral radiographs after placement of the C1-C2 transarticular screw. The study of the 46 patients with normal C1-C2 region had shown the mean values of linear and angular measurements to be greater in males than in females, although no significant difference was found between the two groups (P>.05). The mean distance between the anterior-most point of the anterior tubercle and the middle of the ventral cortex of the lateral mass was 6.5+/-1 mm, and the mean transverse angle of the anterior ring relative to the frontal plane was 22 degrees+/-3.1 degrees. Axial CT evaluation of the individual anatomic relationships of the atlas is simple and may be a useful guide in the determination of the length of the transarticular screw when performed during surgery under lateral fluoroscopic control. PMID- 10791588 TI - Ewing's sarcoma of the middle finger in an infant. PMID- 10791589 TI - Intrathoracic paraspinal desmoid with intracanalicular extension: computed tomography and magnetic resonance imaging findings. PMID- 10791590 TI - Obturator internus pyomyositis. PMID- 10791591 TI - The efficacy of school screening for scoliosis. PMID- 10791592 TI - The German Environmental Survey 1990/1992 (GerES II): a representative population study. AB - The German Environmental Survey (GerES) is a large-scale population study which has repeatedly been carried out in Germany. GerES I was conducted in 1985/1986 followed by GerES IIa in 1990/1991 (West Germany) and GerES IIb in 1991/1992 (East Germany). GerES III is currently run in both parts of Germany. The main goal of the surveys is to analyse and document the extent, distribution and determinants of the exposure to environmental pollutants of the German general population. Field work is conducted using a combination of several tools, including questionnaires, interviews, human biomonitoring, and indoor and outdoor environmental samplings. This paper describes the design of GerES II, and gives a general outline of the field work and the analytical procedures used. In GerES II, about 4000 adults were representatively selected from the German population with regard to age, gender and community size. Approximately 700 children were also included. Arsenic, cadmium, copper, chromium, lead, and mercury were determined in blood, morning urine and/or scalp hair. In addition, samples were taken and analysed to characterise exposure in the domestic environment (indoor air, house dust, drinking water). The contribution to exposure of food was also studied. Exposure to volatile organic compounds (VOCs) was determined by personal sampling. PMID- 10791593 TI - The German Environmental Survey 1990/92 (GerES II): sources of personal exposure to volatile organic compounds. AB - In the framework of the second German Environmental Survey carried out in the Western part of Germany in 1990/91 (GerES IIa) 113 adults aged 25-69 years were selected at random from the total study population of about 2500 to investigate personal exposure to about 70 volatile organic compounds (VOCs). Each subject wore a diffusive badge-type sampler for 1 week. The VOCs determined included alkanes, aromatics, aliphatic halocarbons, terpenes, and oxygen-containing compounds. Multivariate regression analysis was carried out to determine and quantify the major sources of personal exposure to various VOCs. In this paper, results are given for benzene, and C8- and C9-aromatics. Being subject to environmental tobacco smoke was found to be the most important determinant of benzene exposure, but automobile-related activities such as driving a car or refuelling, were also associated with significantly increased levels of benzene. The major determinant of C8- and C9-aromatics concentrations was occupational exposure. Emissions from paints, lacquers, newspapers, magazines and print-works were also important contributors to C8-aromatics exposure. Renovation, painting and smoking were associated with a significant increase of the exposure to C9 aromatics. PMID- 10791594 TI - The German Environmental Survey 1990/1992 (GerES II): cadmium in blood, urine and hair of adults and children. AB - As a follow-up of an earlier population study carried out in West Germany in 1985/1986 (GerES I), a nationwide Environmental Survey was conducted in Germany in 1990-1992 (GerES II). It was the aim of these studies to obtain representative data on the population's body burden and the quality of their indoor environment and immediate surroundings. The present paper reports on cadmium levels in blood, urine and hair of 4021 adults aged 25-69 and 736 children aged 6-14. The statistical analysis included both descriptive and inferential methods. The data were classified according to social factors, lifestyle characteristics, and environmental exposures. Moreover, regression analysis was used to determine the predictors of cadmium concentrations in the types of sample studied. Active cigarette smoking was found to be dominant in affecting blood and urine cadmium levels in adults, but less important for cadmium levels in hair. Age and creatinine level in urine were additional important factors influencing the cadmium concentration in urine, especially in women. Environmental and occupational exposures to cadmium played only a minor role in the exposure models for German adults. The cadmium concentration in blood and urine was generally lower in children than in adults, while no significant difference for cadmium in hair could be detected. The cadmium concentrations in all three samples were significantly higher in East German children than in West German children. PMID- 10791595 TI - Assessment of indoor fine aerosol contributions from environmental tobacco smoke and cooking with a portable nephelometer. AB - Personal monitoring studies have indicated that environmental tobacco smoke (ETS) and cooking are major indoor particulate sources in residential and nonindustrial environments. Continuous monitoring of fine particles improves exposure assessment by characterizing the effect of time-varying indoor sources. We evaluated a portable nephelometer as a continuous monitor of indoor particulate levels. Simultaneous sampling with the nephelometer and PM2.5 impactors was undertaken to determine the relationship between particle light scattering extinction coefficient (sigma(sp)) and particle mass concentration in field and environmental chamber settings. Chamber studies evaluated nephelometer measurements of ETS and particles produced from toasting bread and frying foods. Field measurements were conducted in 20 restaurants and bars with different smoking restrictions, and in five residential kitchens. Additional measurements compared the nephelometer to a different mass measurement method, a piezobalance, in a well-characterized residence where various foods were cooked and ETS was produced. Since the piezobalance provides 2-min average mass concentration measurements, these comparisons tested the ability of the nephelometer to measure transient particle concentration peaks and decay rate curves. We found that sigma(sp) and particle mass were highly correlated (R2 values of 0.63-0.98) over a large concentration range (5-1600 microg/m3) and for different particle sources. Piezobalance and gravimetric comparisons with the nephelometer indicated similar sigma(sp) vs. mass slopes (5.6 and 4.7 m2/g for piezobalance and gravimetric comparisons of ETS, respectively). Somewhat different sigma(sp) vs. particle mass slopes (1.9-5.6 m2/g) were observed for the different particle sources, reflecting the influence of particle composition on light scattering. However, in similar indoor environments, the relationship between particle light scattering and mass concentration was consistent enough to use independent nephelometer measurements as estimates of short-term mass concentrations. A method to use nephelometer measurements to determine particulate source strengths is derived and an example application is described. PMID- 10791597 TI - Pesticide storage and use patterns in Minnesota households with children. AB - As part of the National Human Exposure Assessment Survey (NHEXAS), residential pesticide storage and use patterns were evaluated in a population-based sample of Minnesota households with children aged 3-13. In-home interviews and inventories were conducted to identify pesticide products stored and used in and around 308 households. This statistically based sample represents more than 49,000 urban and rural households in the census tracts sampled. More than 850 unique products were identified using Environmental Protection Agency (EPA) registration numbers. Pesticide products were found in 97% and reported used in 88% of study households. Population-weighted mean values for pesticide storage and use were 6.0 and 3.1 products per household, respectively. The most common active ingredients found were diethyl toluamide (DEET) and related compounds, piperonyl butoxide, pyrethrins, dimethylamine 2-[2-methyl-4-chlorophenoxyl propionate (MCPA) and chlorpyrifos. Household socio-demographic characteristics explained little of the variability in pesticide storage and use patterns, and there were no significant differences in residential storage and use patterns between households located in urban versus non-urban census tracts. Although the prevalence of households with pesticide products was similar to recent national surveys, observed storage and use rates were almost twice those obtained in recent national studies, reflecting improved inventory techniques used by this study and/or increased rates of pesticide presence and use in study households. PMID- 10791596 TI - Design strategy for assessing multi-pathway exposure for children: the Minnesota Children's Pesticide Exposure Study (MNCPES). AB - Although children are exposed to a variety of environmental hazards, including pesticides, there is a scarcity of information available to estimate exposures realistically. This article reports on one of the first attempts to measure multi pathway pesticide exposures in a population-based sample of urban and non-urban children. A design strategy was developed to assess multi-pathway pesticide exposures in children using personal exposure measurements in combination with complimentary measurements of biological markers of exposure, concentrations in relevant environmental media, and time spent in important microenvironments and participating in exposure-related activities. Sample collection and analysis emphasized measurement of three insecticides (i.e., chlorpyrifos, diazinon, and malathion) and one herbicide (i.e., atrazine). These compounds were selected because of their frequent use, presence in multiple environmental media, expected population exposures, and related hazard/toxicity. The study was conducted during the summer of 1997 in Minnesota and involved a stratified sample of households with children ages 3-12 years. Participants resided in either (a) the cities of Minneapolis and St. Paul (urban households), or (b) Rice and Goodhue Counties just south of the metropolitan area (non-urban households). Results from a residential inventory documenting storage and use of products containing the target pesticides were used to preferentially select households where children were likely to have higher exposures. The study successfully obtained pesticide exposure data for 102 children, including measurements of personal exposures (air, hand rinse, duplicate diet), environmental concentrations (residential indoor/outdoor air, drinking water, residential surfaces, soil), activity patterns (obtained by questionnaire, diary, videotaping), and internal dose (metabolites in urine). PMID- 10791598 TI - Spatiotemporal analysis of environmental exposure-health effect associations. AB - The goal of this work is to discuss a general methodology for studying associations between environmental exposures and health effect by means of the spatiotemporal random field theory. This theory is the tool of choice for rigorously accounting for important spatiotemporal variations and uncertainties related to exposures and effect. Within the framework of the random field theory, the Bayesian maximum entropy model neatly synthesizes various sources of physical and epidemiological knowledge into spatiotemporal analysis. Therefore, unlike technical statistics, this approach relies on the blending of substantive physical knowledge with powerful mathematical techniques and a coherent rationale. Given the well-founded fact that certain health effects may be caused by environmental exposures, the significance of these exposures is assessed in terms of a criterion that is based on the joint stochastic representation of exposure and health-effect distributions in space/time. In view of this criterion, the strength and consistency of the exposure-effect association are evaluated on the basis of the health-effect predictions that the combined physico epidemiologic analysis generates in space/time. The main features of the approach are demonstrated by a simulation example and a real case study involving mortality and cold temperatures in North Carolina. The studies demonstrated the practical usefulness of the stochastic human exposure analysis in assessing the exposure-effect association. The results reported here emphasize the links between spatiotemporal models of physical systems and population health-effect distributions, thus suggesting directions for improving the current understanding of quantitative "exposure-health effect" functions. PMID- 10791599 TI - Sources of nitrate exposure in residents of rural areas in Quebec, Canada. AB - Nitrate exposure was investigated in a group of 187 people using well water and living in four areas of rural Quebec (Canada) with intensive agricultural activities. Nitrate intake was evaluated using a 24-h dietary recall and a food frequency questionnaire, in conjunction with a validated food database and measurements of nitrate concentrations in private wells. The total internal dose was estimated by means of the 24-h urinary nitrate excretion, while taking into account risk factors for endogenous nitrate formation. Mean (geometric) 24-h urinary nitrate excretion was 16.9 mg N for the 100 people with low groundwater contamination (mean nitrate concentration=0.18 mg N/l) and 23.3 mg N in the 87 individuals with moderate groundwater contamination (mean nitrate concentration=7.1 mg N/l). A multivariate analysis revealed that dietary nitrate intake during the last 24 h was the principal source of exposure, followed by water intake during the last 24 h. The Quetelet index was also a significant predictor of urinary excretion. The total predictive model explained only 29% of the variability in urinary nitrate excretion (R2=0.286). Neither the inflammatory status as indicated by elevated C reactive protein, the presence of Helicobacter pylori antibodies nor the occurrence of diarrhea during the last 24 h prior to urine collection were associated with urinary nitrate excretion. In conclusion, food and to a lesser extent water contribute to nitrate exposure in this rural setting with moderate water contamination. Better predictors of endogenous nitrate production are needed to improve our ability to model nitrate body burden and estimate associated health risks. PMID- 10791601 TI - A comparison of recent and long-term average measurements of nitrate in drinking water. AB - We evaluated the usefulness of a recent measure of drinking water nitrate as a predictor of long-term average nitrate exposure calculated from historic data. Exposure estimates were calculated for 214 study participants who used public water supplies between 1947 and 1980 in Minnesota. Long-term average nitrate was calculated by linking residential histories to historical nitrate data. For recent exposures, we averaged nitrate measurements in 1980, or the next closest year with measurements. The Spearman correlation coefficient for the relationship between the two measures was 0.54 (95% confidence interval [CI]=0.44-0.63). Agreement was highest among those residing 34 or more years in their town as of 1980 (r(s)=0.70; 95% CI=0.55-0.80). These findings suggest that taking into account the study participants' duration of residence may enhance the validity of using a recent measure as an indicator of past exposures. PMID- 10791600 TI - Longitudinal investigation of exposure to arsenic, cadmium, chromium and lead via beverage consumption. AB - Semi-quantitative food checklists and duplicate beverage samples were collected from up to 80 individuals in Maryland in 1995-1996 in as many as six approximately equally spaced sampling cycles as part of a pilot longitudinal exposure investigation. The duplicate beverage samples were homogenized and analyzed for arsenic (As), cadmium (Cd), chromium (Cr) and lead (Pb) using inductively coupled plasma mass spectrometry (ICP-MS). Metal concentrations (microg/kg) and weights of the duplicate beverage samples (kg/day) were used to derive average daily exposure (microg/day) for each metal. Mixed models and generalized linear models were used to evaluate temporal and population variability of the beverage consumption rates, the log-transformed metal concentrations in the beverage samples, and the associated exposures. The mean number of beverage servings consumed per day was 3.4 (SD 1.9). The temporal variability of the total beverage consumption rates was found to be significant (p = 0.0476). As, Cd, Cr, and Pb were present at quantifiable levels in 93.5, 76.0, 93.5, and 96.7% of the beverage samples, respectively. The mean concentration in the samples was 2.0 (SD 4.4) microg/kg for As, 0.9 (1.6) for Cd, 29.2 (138.5) for Cr, and 2.0 (2.4) for Pb. The mean log-transformed concentrations for As, Cr and Pb and exposure for As varied by as much as a factor of 3 across sampling cycles and were statistically significantly different (p<0.05). Concentrations and exposures of all four metals varied significantly among participants. These findings are discussed with respect to the data collection methods, results from comparable studies, and implications for exposure and risk assessment. PMID- 10791602 TI - Learning and creative thinking. PMID- 10791603 TI - Health and disease in social perspective. PMID- 10791604 TI - Early medicine in Mesopotamia. PMID- 10791605 TI - Sexually dimorphic effect of an acute smoking manipulation on skin resistance but not on heart-rate during a cognitive verbal task. AB - In a two-day, two-session experiment where smokers male and female college student subjects worked on a cognitive verbal task during either the first or second day, and on a cognitive spatial task on the second or first day, smoking was manipulated as an acute independent variable by requiring 10+ hours of pre experimental abstention, and providing a cigarette during the 15-minute rest period between the two sessions. Non-smoker female and male subjects underwent the same experiment, and hence served as controls for the effects of this acute smoking manipulation. Overall adaptation (decreased arousal) to the experiment was manifested in a significant increase in skin resistance level (SRL) in all subjects, but when this adaptation effect was statistically controlled, there was a significant smokers by sex interaction during the verbal task only, such that SRL was increased by the cigarette in males, but decreased in females. In contrast, the same analysis indicated only a marked increase in heart-rate (HR) due to smoking, which was unaffected either by sex or by whether the task was the verbal or the (easier) spatial one. We interpret the SRL results as reflecting a sex difference in the direction of transient psychological arousal, and discuss it in relation to evidence in the literature based on self reports, and to evidence (based on HR in this study and on blood pressure in other studies) on physiological (cardiovascular) arousal. Key Words: Electrodermal activity, heart rate, psychological vs. physiological, verbal and spatial cognitive tasks. PMID- 10791606 TI - Gender differences in acute and chronic stress in Wistar Kyoto (WKY) rats. AB - While females are considered more susceptible to depressive behavior, this assertion is not strongly supported by the experimental literature. Since stress contributes to depressive behavior, male and female Wistar Kyoto (WKY) rats were exposed to either one session (acute stress) or 5 sessions (chronic stress) of restraint plus cold in order to study depressive behavior in male and female rats. After their respective treatment exposure, rats were tested in the open field test (OFT) and for retention of a passive-avoidance (P-A) task. One stress session resulted in significant immobility in the OFT for males, whereas 5 sessions were required to produce similar immobility in female rats. Acute stress interfered with the retention of the P-A response for males, while both acute and chronic stress produced poor P-A responses in female rats. Food consumption decreased progressively, as a function of stress sessions, in female rats, whereas feeding in males returned to control levels after five stress days. Both acute and chronic stress exacerbated the stress ulcer response in male rats, but not in female rats. Chronic, but not acute, stress resulted in an increase in serotonin transporter mRNA levels in the dorsal raphe nucleus of both male and female rats. The general consensus from these data suggested that female rats were more vulnerable to chronic stress and consequently supported the notion that females may be more susceptible to stress-induced behavioral depression. Key Words: WKY rats, acute and chronic stress, gender, passive avoidance, open field behavior, stress-ulcer, adrenal weight, serotonin, dorsal raphe nucleus PMID- 10791607 TI - Some hazards of invasive cardiology. AB - Since the introduction of cardiac catheterization by Andre Cournand and Dickinson Richards, the valuable diagnostic and therapeutic device has encouraged many action-minded physicians to use cardiac catheterization to develop a new specialty, invasive cardiology. The data to be presented here derive from a catastrophe that occurred during an invasive treatment of a 54-year-old man who had experienced an ordinary myocardial infarction. PMID- 10791608 TI - The impact of an emotional self-management skills course on psychosocial functioning and autonomic recovery to stress in middle school children. AB - Unmanaged emotional reactions to stress not only lead to behavior problems in young people but also create physiological conditions that inhibit learning and potentially increase the risk of disease later in life. For these reasons, the integration of emotional self-management skills training programs has become an increased priority in some schools. In this study, middle school students enrolled in a course in emotional competence skills learned techniques designed to intercept stressful responses during emotionally challenging situations. Behavioral outcomes were assessed using the Achievement Inventory Measure and autonomic function was measured by heart rate variability (HRV) analysis during and after a stressful interview. Following the program, students exhibited significant improvements in areas including stress and anger management, risky behavior, work management and focus, and relationships with family, peers and teachers. These improvements were sustained over the following six months. Students using the skills taught in the course to recover from acute emotional stress were also able to positively modulate their physiological stress responses. As compared to a control group, trained students demonstrated significantly increased HRV and more rhythmic, sine wave-like heart rhythm patterns during recovery. This response pattern reflects increased parasympathetic activity, heart rhythm coherence, and entrainment of other biological oscillatory systems to the primary heart rhythm frequency. Increased physiological coherence is associated with improved cognitive performance, emotional balance, mental clarity and health outcomes. These physiological shifts could promote the sustained psychological and behavioral improvements associated with the use of emotional management skills. It is suggested that learning emotional competence skills in childhood establishes healthier physiological response patterns which can benefit learning and long-term health. Results provide support for the integration in school curricula of courses designed to teach effective self-management skills to children. PMID- 10791609 TI - Somatovisceral interactions in visceral perception: abdominal masking of colonic stimuli. AB - Clinical and experimental evidence on referred pain and spinal-afferent convergence demonstrates a close relationship between visceral and somatosensory perception, which is important for current models of symptom perception and central body representation. The study uses a psychophysical approach to quantify these interactions at the perceptual level, taking into account problems of comparable intermodal scaling and the role of awareness. An experiment on somatosensory masking of distension stimuli in the colon is reported in which a multiple staircase method of forced choice discrimination with concurrent sensation ratings was employed. Results showed perceptual masking of visceral by abdominal stimuli but not vice versa. The masking effect was not enhanced by intratomal placement of the abdominal stimulus in the lower left quadrant. This contradicts the spinal sensory convergence model and points to perceptual interactions at higher brain levels. Loglinear analysis of relations between discrimination and subjective sensation revealed qualitative differences of somatovisceral perception at the preconscious as compared to the conscious level. This argues for a two-process model of integrative body perception. PMID- 10791610 TI - The relationship between external events and affect states in older people. AB - Long-term and moderately short-term effects of bereavement and marriage on psychological well-being (PWB) among older people were investigated. The aspect of PWB that was examined was the prevalence of six affects, rated in terms of their frequency during the past year. Affect frequency of four groups was tested: Recently widowed, recently married, and widowed and married elders unselected for length of time in those marital statuses. As predicted, both length of time in the marital status and congruence between the positive event (marriage) and positive affect and between congruence of the negative event (bereavement and negative affect) were associated with group differences. Depressive affect was greatest among the recently bereaved but the recently-married, long-married, and longer-bereaved groups did not differ in depression. Positive affect was greatest among the recently married and other groups did not differ in this respect. Hostility, anxiety, shyness, and contentment were not predicted to differ among groups; in fact, contentment was least in the bereaved; shyness was least among the recently-married, and hostility was lowest among the long-widowed. Results are discussed in terms of the joint influences of time since a life event and the differential relevance of positive and negative affect states to positive and negative events. Continued research attention to the covariation of these factors in relation to the affective aspects of PWB is needed to understand the conditions of stability and change. PMID- 10791611 TI - Coronary catherization patient and wife's perceptions of social support: effects due to characteristics of recipient, provider, and their interaction. AB - This investigation explored relationships between coronary catherization patient and wife's characteristics and their perceptions of social support. Participants were 124 male patients undergoing diagnostic catherization to detect coronary artery disease (CAD) and their wives. Patients and wives' mean ages were fifty nine and fifty-six years, respectively. While visiting the hospital both patient and wife independently completed a brief questionnaire that assessed psychological and physical health characteristics and perceptions of social support. Congruent with the interdependent and overlapping contexts, and reciprocal relationships assumptions of Revenson's (1994) ecological/contextual framework, results suggested characteristics of patients and wives (e.g., age, mental health, social functioning, hostility, depression, general perceived health) to interactively moderate their perceptions of social support. Overall, perceptions of social support were found to be influenced by characteristics of the perceiver, the support provider, and their interaction. Concerns for supportive and helpful caregiving are discussed. PMID- 10791612 TI - Derivation and pilot assessment of a health promotion program for Mandarin speaking Chinese older adults. AB - As the percentage of older adults of diverse ethnicities increases in the United States, the call for culturally sensitive health care service strategies that target the special needs of older people grows. The present report describes methods used to adapt a health care program so that it would better meet the needs of a group of well, older Mandarin-speaking Chinese residents of Los Angeles. The specific qualitative research procedures that we used to adapt the treatment program are described, along with the particular adaptations that emerged. Additionally, outcomes from a randomized pilot experiment are presented that are consistent with the notion that the adapted program was effective in reducing health-related declines among older Mandarin-speaking men and women. The overall outcome of this project is in agreement with other reports in the health care literature that address the importance of providing culturally sensitive health care service for elders. PMID- 10791614 TI - Rheumatology and the Bone and Joint Decade 2000-2010 ILAR UMER 2000 project. International League of Associations for Rheumatology. Undergraduate Medical Education in Rheumatology. PMID- 10791613 TI - The extended effects of the life review in nursing home residents. AB - To date, there is little information on the therapeutic effects of the life review beyond one year. This analysis followed fifty-two of 256 subjects who lived for at least three years in a nursing home. These participants received either a life review or friendly visit and took part in four repeated testings to determine the lasting effects of the life review at two and three years. Measures of integrity (life satisfaction, psychosocial well being, self-esteem) and despair (depression, hopelessness, and suicide intent) were used as pretest, posttest, and retest. Results showed a trend toward continued and by year three significant improvement over time in those who received the life review on measures of depression (t = -2.20, p < .03), life satisfaction (t = 2.51, p < .02), and self-esteem (t = -2.31, p < .03). PMID- 10791615 TI - The outcome of knee synovitis in early arthritis provides guidelines for management. AB - The aim of the study was to examine the clinical outcome of patients presenting to an early arthritis clinic with synovitis of the knee. The patients were assessed at presentation for evidence and pattern of joint inflammation. These patients were then reassessed at 3, 6 and 12 months and thereafter annually to determine clinical outcome. One thousand six hundred and thirty-three consecutive referrals were examined, 903 of whom had early synovitis. One hundred and thirty had knee synovitis at presentation, of whom 73 fulfilled ACR criteria for rheumatoid arthritis (RA) during the study. All 73 presented with a symmetrical polyarthritis that included the small joints and had persistent disease at 1 year. Of the remaining 57 patients, 61% of those presenting with an oligoarthritis and 33% with a polyarthritis (including knee synovitis) were in remission at 1 year. None of those presenting as a monoarthritis of the knee had inflammation at 1 year or fulfilled ACR criteria for RA at any time. It was concluded that patients presenting with knee synovitis in the absence of a small joint polyarthritis usually have a benign course following standard therapy. No patient who presented with monoarthritis developed RA. Knee synovitis as part of a polyarthritis (even when not fulfilling ACR criteria) probably justifies disease-modifying antirheumatic drug at presentation. PMID- 10791616 TI - Avascular areas on nailfold capillary microscopy of patients with Wegener's granulomatosis. AB - Over the past 25 years, nailfold capillary microscopy (NCM) has gained diagnostic value in the field of rheumatology based on descriptive data from patients with distinct connective tissue diseases (CTD). We prospectively analysed NCM findings from 116 patients selected for NCM by one of the following indications: (1) suspected diffuse or limited scleroderma or dermatomyositis, (2) evaluation of Raynaud's phenomenon (RP) or (3) suspected small-vessel vasculitis. Nailfold haemorrhages, and enlarged and tortuous nailfold capillaries ('lupus pattern') were found to comparable degrees in patients with CTD and primary RP. Only giant loops, bushy capillaries and avascular areas indicated CTD; 92% of patients with Wegener's granulomatosis (WG) had avascular areas. From all nailfold capillary abnormalities, only bushy capillaries, giant loops and avascular areas support a suspected CTD. A lupus-like pattern is not diagnostic. Avascular areas are a typical abnormality in patients with WG, for which NCM findings have not been described previously. PMID- 10791617 TI - Tissue concentration of the active substance of Voltaren SR 75 in articular cartilage, synovial membrane and bone. AB - The tissue concentration of diclofenac sodium, the active substance of Voltaren SR 75 mg tablets, was studied in 10 patients with coxarthrosis. Voltaren SR 75 mg tablets were administered twice daily for 5 days, then prosthetic joint replacement of the hip was performed. Tissue levels of diclofenac were determined by HPLC in specimens of articular cartilage, synovial membrane and bone obtained during surgery. The results demonstrate that diclofenac was present in all three tissue types after 5 days of therapy. PMID- 10791618 TI - Biochemical bone turnover markers in patients with ankylosing spondylitis. AB - In this study, bone formation markers [alkaline phosphatase (ALP), bone-specific alkaline phosphatase (BALP) and osteocalcin (BGP)] and bone resorption markers [pyridinium cross-links: pyridinoline (Pyd) and deoxypyridinoline (Dpyd)] were studied in 44 patients with ankylosing spondylitis (AS) and in a control group of 41 subjects. The AS group was classified according to duration of disease, erythrocyte sedimentation rate (ESR) values and gender. Urinary Pyd and Dpyd concentrations of the patients were higher than in the control group, concomitant with the lower T-score values of the patients in both the anteroposterior lumbar spine and femoral neck. Although a correlation between markers of disease activity [ESR and C-reactive protein (CRP)] and bone turnover markers was not observed, urinary excretion of Dpyd and Pyd was enhanced in the ESR >20 mm/h subgroup compared with the ESR < or =20 mm/h subgroup. A significant elevation of urinary Dpyd was also observed in the female subgroup and long disease duration subgroup. Serum ALP, BALP and BGP levels of the patients and control group were not statistically different (p>0.05). No significant differences were observed between mineral and calcitropic hormone levels in either group, and total testosterone levels of the patients were within the reference range. According to this study, urinary Pyd levels are elevated in patients with AS. Gender, duration of disease and ESR also have an impact on urinary excretion of these collagen compounds. It can be concluded that bone turnover in patients with AS reflects normal osteoblastic activity and high osteoclastic activity. PMID- 10791620 TI - Maximal isometric muscle strength: normative values and gender-specific relation to age. AB - To date, there have been very few studies on the age dependence of maximal isometric muscle strength (MIMS) in healthy subjects aged 20-80 years, based upon measurements of a large number of functional muscle groups (FMGs). Using a hand held pull gauge it is possible to measure MIMS of nearly every FMG. The objectives of this study were to obtain normative values for MIMS, to evaluate differences in MIMS in relation to gender and body side and to compare the age dependence of muscle strength between women and men. In a convenience sample of 290 healthy women (aged 20-82 years) and 253 men (aged 21-79 years), MIMS of 51 FMGs was measured. For each FMG the age dependence of MIMS was depicted, side and gender specific, as percentile curves and was analysed using linear quantile regression analysis. MIMS was found to be significantly higher in men than in women and higher on the right than on the left side. A biphasic model with linear equations for strength medians was derived for each gender. The age at transition from phase 1 to phase 2 was 55 years (SD 8) for women and 49 years (SD 13) for men. During phase 1, MIMS did not decrease significantly. During phase 2, MIMS decreased in all FMGs in both genders with a steeper slope in women (-0.92) than in men (-0.63). The age dependence of MIMS differed significantly between women and men. The present study gives gender-specific equations which enable one to calculate normative values for MIMS, as measured with a pull gauge, based upon age. These normative values will allow an objective assessment of patients with diminished muscle strength as, for example, in myositis, rheumatoid arthritis and nerve root compression syndromes or in the elderly. PMID- 10791619 TI - Oxaceprol is a well-tolerated therapy for osteoarthritis with efficacy equivalent to diclofenac. AB - The therapeutic equivalence and safety of treatment for 21 days with 400 mg t.i.d. oxaceprol (n = 132) and 50 mg t.i.d. diclofenac (n = 131) were assessed in a multicentre, randomised, double-blind study of a mixed population of patients with osteoarthritis of the knee and/or hip. In a per-protocol analysis of efficacy, the mean Lequesne index decreased by 2.5 points in the oxaceprol group (n = 109) and by 2.8 points in the diclofenac group (n = 109). The 95% confidence interval for the end-point difference revealed therapeutic equivalence. This was confirmed by assessments (visual analogue scale) of pain at rest, weight-bearing pain, pain on standing and pain on movement, all of which decreased to a similar extent under both treatments. The pain-free walking time increased in both groups from 10 min to 25 min by the end of the treatment period. Mobility was also increased to a similar extent by both drugs. The physicians assessed treatment as good or very good in 45-46% of patients in both groups. In all patients who received treatment, 28 and 37 adverse events were reported by 25 out of 132 (18.9%) and 33 out of 131 (25.2%) patients treated with oxaceprol and diclofenac, respectively. In 15 patients (11.4%) with 15 adverse events in the oxaceprol group and 25 patients (19.1%) with 27 adverse events in the diclofenac group, a relation to the medication was considered probable. The difference between the groups was statistically significant (p = 0.04106) for the number of these adverse events. Oxaceprol is therapeutically equivalent to diclofenac, but better tolerated than diclofenac in the treatment of osteoarthritis. PMID- 10791621 TI - Efficacy of methotrexate in the treatment of ankylosing spondylitis: a three-year open study. AB - The aim of the study was to evaluate the efficacy of methotrexate treatment in patients with ankylosing spondylitis in a 3-year open trial. Seventeen patients, 14 men and three women (mean age 32.7+/-8.9 years), suffering from ankylosing spondylitis and non-responders to treatment with sulphasalazine, were enrolled in our study. Sixteen of them were evaluable at the end of the study. Methotrexate (7.5-10 mg/week) was administered for 3 years. Efficacy was evaluated on the basis of clinical and laboratory variables, radiographic signs of disease progression and daily dosage of indomethacin. We obtained a good and relatively prompt clinical response except for peripheral arthritis and iridocyclitis; in fact, after 3 months of methotrexate treatment a significant amelioration of the following parameters was observed: visual analogue scale for the evaluation of both night pain and general well-being, Shober's test, occiput-wall distance, fingertip to floor, erythrocyte sedimentation rate, C-reactive protein level and daily dose of indomethacin. A further improvement was obtained during the subsequent period. Radiographs of the spine and sacroiliac joints did not show any signs of disease progression. Side-effects were a transitory elevation of transaminases (four cases) and slight hypogammaglobulinaemia (one case). Methotrexate treatment may be useful in ankylosing spondylitis, but a combined treatment might be indicated for patients with peripheral arthritis. PMID- 10791622 TI - Coeliac disease in spondyloarthropathy: usefulness of serological screening. AB - The aim of our study was to find unidentified or neglected cases of coeliac disease by using serological screening in a group of patients with spondyloarthropathies. Altogether, 74 consecutively hospitalised patients (28 females, 46 males, from 15 to 72 years of age, mean age 40.4+/-1.6 years) with spondyloarthropathies were investigated by serological screening tests for coeliac disease. IgA- and IgG-type antigliadin antibodies were determined using an enzyme-linked immunosorbent assay, and IgA- and IgG-type antireticulin and IgA type antiendomysium antibodies were measured by an indirect immunofluorescence method. An increased level of antigliadin antibodies was found in nine (12%) of the studied patients and in one of them, antiendomysium antibodies were revealed. In this HLA B8-positive patient, typical villous atrophy with crypt hyperplasia was found in the small bowel biopsy specimen, which confirmed the diagnosis of coeliac disease. None of the patients had IgA- and IgG-type antireticulin antibodies. We found an association of spondyloarthropathy with coeliac disease in one patient out of 74. Clinicians need to be aware of this association, which has important implications for the correct management of patients. PMID- 10791623 TI - Anticentromere antibody as a risk factor for cancer in patients with systemic sclerosis. AB - This study has estimated the cancer risk among patients with systemic sclerosis (SSc) using a population-based analysis. Using the inpatient and outpatient registries for patients at Kyushu University Hospital between 1982 and 1996, standardised incidence rates (SIRs) (ratio of observed-to-expected cancers) were calculated in 43 patients with SSc, 24 patients with polymyositis (PM) and 17 patients with dermatomyositis (DM). Risk factors predisposing to cancers were also investigated in the SSc patients. Compared with the Japanese general population, the SIR for developing cancer in SSc patients was 5.1 (95% confidence interval (CI), 1.7-10.8), while the SIRs for cancer in the PM and DM groups were 4.7 (95% CI, 1.5-10.3) and 61.2 (95% CI, 46.8-77.6), respectively. A statistically significant risk factor for cancers in the SSc patients was positivity for anticentromere antibody (ACA) (p<0.05), while the erythrocyte sedimentation rate, serum lactate dehydrogenase concentration, serum gamma globulin concentration, titre of antinuclear antibody and positivity for antitopoisomerase I antibody were not associated with cancer in SSc. Our population-based study confirms the increased risk of cancer among patients with SSc in Japan and provides new evidence that positivity for ACA should be considered as a risk factor for cancer in future monitoring of patients. PMID- 10791624 TI - Different articular outcomes of Still's disease in Chinese children and adults. AB - The clinical manifestations, treatment and course, and articular outcomes of 24 children with juvenile-onset Still's disease (JOSD) and 21 adults with adult onset Still's disease (AOSD) were compared retrospectively. There was no significant difference in the initial clinical and laboratory manifestations except that more adults presented with a sore throat (81% vs. 46%, p = 0.03). Although serum ferritin was almost always elevated in both diseases, adults had significantly higher serum ferritin concentrations compared with those of children. Steroid treatment was required in 71% of children and 52% of adults, while disease-modifying antirheumatic drugs were used in 42% of children and 24% of adults during the course. Chronic arthritis (>6 months) occurred in comparable proportions of patients with JOSD and AOSD (46% vs 38%, p = 0.82), irrespective of the disease pattern (monocyclic or polycyclic). However, severe deforming arthritis with marked functional limitation occurred only in JOSD, particularly with polyarthritis at disease onset (more than five affected joints). In contrast, AOSD patients with chronic arthritis had a favourable functional outcome at the end of the follow-up. Our study suggested different articular outcomes of Still's disease in Chinese children and adults. PMID- 10791625 TI - Clinical assessment of spinal mobility measurements in ankylosing spondylitis: a compact set for follow-up and trials? AB - Different spinal ranges of motion (ROM) were measured and the results of 17 repeated tests correlated with spinal radiological changes in 52 male patients with ankylosing spondylitis (AS). Both Schober tests and measurements of lumbar and cervical rotations (TRi, TR, CR, CRt) and lateral flexions (LFLf, LFLx, CLFLt, CLFLm), together with thoracolumbar flexion (ThFL), cervical flexion extension measurements (CFL, CExt), and tragus - wall and occiput - wall distances (OWD,TWD), showed significant correlations with detailed radiological spinal changes. Cervical rotation (CRm, CRt) and flexion (CFLm) correlated only with cervical changes, and thoracolumbar rotation as assessed by instrument (TRi) correlated only weakly with lumbar changes, while chin-chest distance (CCD) and chest expansion (CE) showed no correlation. Inter- and intratester reliability was good in all tests (the intraclass correlation coefficients ranged from 0.84 to 0.98). Three new tape methods for measuring thoracolumbar and cervical rotations and cervical lateral flexion also proved to be valid and reliable, as did the Schober-S1 modification. We conclude that the thoracolumbar segment (Schober), whole (ThFL) and lateral (LFL) flexions and rotation (TR), and chest expansion (CE) (after careful standardisation) together with cervical rotation (CR), extension (CExt) and/or lateral flexion (CLFL) comprise the set of mobility tests for the follow-up and assessment of disease progression in AS. On the other hand, cervical (forward) flexion (CFL), chin-chest distance (CCD) and an instrument method for thoracolumbar rotation (TRi) are not approaches to be recommended. PMID- 10791626 TI - Efficacy of early treatment of severe juvenile dermatomyositis with intravenous methylprednisolone and methotrexate. AB - A pilot study was conducted to assess the efficacy of early treatment of severe juvenile dermatomyositis (JDMS) patients with intravenous methylprednisolone (IVMP) and methotrexate (MTX). Twelve children diagnosed with severe JDMS were treated with IVMP and MTX. Six patients were treated early (within 6 weeks of the diagnosis) while in the other six patients, MTX was started 5-72 months after the diagnosis was made. The clinical responses of the patients to treatment, including alterations in muscle strength, muscle enzyme levels and corticosteroid dosage as well as the development of side-effects, were recorded. The indications for starting the treatment were defined and documented. The primary measures of response were resolution of the clinical indications for treatment, decreased activity of the disease manifestations and tapering of the corticosteroids to the minimal dose or discontinuation without clinical or biochemical flare. The main indications for starting IVMP and MTX were dysphagia and severe cutaneous vasculitis. All the patients received MTX orally for at least 8 months, as well as IVMP (30 mg/kg/dose), but none of the patients was on additional second-line treatments. The six patients who were treated early with MTX showed a significant clinical improvement. In five out of the six, the corticosteroid dosage was eventually reduced to <5 mg/day. None of them developed calcinosis. In contrast, two of the six patients who were treated late with MTX developed calcinosis. This study suggests that MTX and IVMP are a useful combination in the early treatment of severe JDMS. Given the fact that our sample was small, further studies in a controlled trial are necessary to confirm these findings. PMID- 10791627 TI - Clinical manifestations of Korean female gouty patients. AB - In an endeavor to analyse the clinical characteristics of female gout, we reviewed 36 women with gout. Twenty-seven (75%) developed the first symptomatic episode of gout after the onset of the menopause. The mean age at onset of gout was 54.3 years (range 15-87 years). Twenty-two patients (61%) had hypertension, 17 (47%) had renal insufficiency, 13 (36%) used diuretics and 10 (28%) were taking cyclosporine for a renal allograft. Tophaceous gout occurred in 10 patients (27%) and polyarticular involvement was seen in 16 (44%) at initial presentation. Five of nine premenopausal patients were taking cyclosporine and four had renal insufficiency. A comparison with a control group of 72 randomly selected male patients with gout showed that the female patients were frequently receiving diuretics at the time of the attack and had significantly lower mean uric acid excretion, whereas significantly more male patients showed heavy alcohol consumption and precipitating events for an acute attack compared with the female patients. There were no significant differences between the sexes for onset age, hypertension, renal insufficiency, distribution of joint involvement, tophi and mean serum uric acid concentration. The female patients in this study had a lower mean age at onset of gout than in previous studies, which was attributed to the inclusion of renal transplantation patients. Transplantation gout patients receiving cyclosporine lower the mean age at onset of female gout and this is an emerging problem in female gout. PMID- 10791628 TI - Induction of an acute attack of calcium pyrophosphate dihydrate arthritis by intra-articular injection of hylan G-F 20 (Synvisc). AB - Little is known about the induction of acute calcium pyrophosphate dihydrate arthritis after the intra-articular injection of hylan G-F 20 (Synvisc). Two reports have documented this adverse effect after the intra-articular injection of hyaluronan. Our patient, a 60-year-old man with osteoarthritis in both knees, presented with a history of an arthroscopy with meniscus shaving 7 years previously. He was given an injection of hylan G-F 20 in the right knee joint. Two days after the second injection, pain and swelling of the knee occurred. There was a severe loss of physical function. Systemic inflammatory reactions such as fever were not observed. A microscopic investigation of the synovial fluid showed evidence of calcium pyrophosphate dihydrate crystals. Bacterial contamination was not detected. There was no indication for calcium pyrophosphate dihydrate in the history of the patient. Some days after receiving nonsteroidal anti-inflammatory drugs and an intra-articular injection of steroids, the symptoms disappeared. PMID- 10791629 TI - Bilateral hip arthroplasty for ochronotic arthropathy. AB - Ochronosis is a musculoskeletal manifestation of alkaptonuria, a rare hereditary metabolic disorder characterised by the absence of the enzyme homogentisic acid oxidase and associated with various systemic abnormalities related to the deposition of homogentisic acid pigment (ochronotic pigment). In this report, we describe a 53-year-old, HLA-B27(+) woman with ochronotic arthropathy. In addition to the typical clinical features of the disorder, she had bilateral hip involvement, which was improved by cementless total hip prosthesis. PMID- 10791630 TI - Charcot joint in idiopathic sensorimotor neuropathy. AB - Charcot joint or neuropathic arthropathy is described in certain neurological conditions. We report the case of a man who presented with a swollen ankle 10 days after a long walk, which rapidly progressed to a Charcot joint. A neurological examination revealed areflexia and insensitivity to temperature and pain. Electromyographic analysis showed a mixed sensorimotor polyneuropathy. Besides axonal loss and demyelinisation on sural nerve biopsy, prominent loss of unmyelinated fibres was demonstrated. Despite extensive investigations, no definite cause for this neuropathy could be found. PMID- 10791631 TI - Involvement of the entire spinal cord and medulla oblongata in acute catastrophic onset transverse myelitis in SLE. AB - A 30-year-old Caucasian male with systemic lupus erythematosus suffered acute catastrophic-onset transverse myelitis. Two years earlier aseptic meningitis, another rare CNS lupus manifestation, had been diagnosed. MRI showed involvement of the medulla oblongata and the entire spinal cord. Therapy with intravenous high-dose methylprednisolone and intravenous cyclophosphamide is discussed. PMID- 10791632 TI - Periaortitis and aortic dissection due to Wegener's granulomatosis. AB - We describe here a patient with abdominal periaortitis and intramural dissection as early manifestations of Wegener's granulomatosis (WG). Surgical biopsies taken from the retroperitoneal inflammatory tissue surrounding the aorta showed granulomatous vasculitis. The patient had antiproteinase-3 antibodies and suffered from nasal, pulmonary, nervous and renal WG involvement. Although being a vasculitis of medium size and small vessels, WG should be included in the systemic vasculitides which can give rise to (peri)aortic inflammation. PMID- 10791633 TI - Acute-phase proteins in patients with systemic sclerosis. AB - Acute-phase proteins were determined in serum of 20 women with systemic sclerosis and 10 age-matched healthy controls. All the patients had had symptoms of the disease less than five years. An increase in only a few proteins (haptoglobin, alpha1-acid glycoprotein, complement component C3 and alpha2-macroglobulin) was found. The results indicate for the impaired acute phase response in patients with systemic sclerosis. PMID- 10791634 TI - Psoriatic arthritis exacerbated by Salmonella infection. AB - Psoriatic arthritis (PsA) is an inflammatory joint disease in which environmental factors, particularly trauma and infections, are thought to play an important role. The authors describe the case of a patient with a mild and long-untreated form of PsA which was severely exacerbated by Salmonella typhimurium infection. This case confirms the importance of infectious agents in the occurrence and course of PsA. PMID- 10791635 TI - Achilles tendinitis as the presentation form of Lofgren's syndrome. AB - Lofgren's syndrome is characterised by bilateral hilar adenopathy arthritis and erythema nodosum. Achilles tendinitis as the presentation form of Lofgren's syndrome is very unusual. Herein we present a case of bilateral achilles tendinitis as the presentation form of Lofgren's syndrome. PMID- 10791636 TI - Trauma care and trauma surgeons. PMID- 10791637 TI - The influence of the timing of surgery on soft tissue complications and hospital stay. A review of 84 closed ankle fractures. AB - BACKGROUND AND AIMS: Open reduction and internal fixation of an extensively swollen ankle may lead to wound closure problems, blistering, wound edge necrosis and infection. Accordingly, internal fixation should be accomplished either before or after the period of critical soft tissue swelling. The object of the study was to investigate if the timing of surgery had any influence upon soft tissue complications and hospital stay. PATIENTS AND METHODS: The clinical course of the first 6 postoperative weeks of 84 closed ankle fractures treated by open reduction and internal fixation were reviewed. Seventeen patients were not operated on early due to lack of operative capacity and were thus operated on after 5 days or more. These patients were compared to the patients operated on within 8 hours (n = 67). The groups were comparable with respect to age, gender and fracture types. RESULTS: Despite a higher incidence of primary soft tissue injuries in the early group, the patients operated on delayed had a higher incidence of wound infections (17.6% vs. 3.0%) and hospital stay was prolonged with 12.4 days compared to early surgery. All wound infections were found in grossly displaced fractures despite adequate closed reduction immediately after arrival in the hospital. CONCLUSIONS: Delayed surgery of closed ankle fractures increases the risk of soft tissue complications and prolongs hospital stay. Immediate surgery is particularly indicated in the severely displaced ankle fracture, and if not achievable, temporary reduction and immobilization is recommended. PMID- 10791638 TI - Reliability of initial chest radiographs in the diagnosis of blunt diaphragmatic rupture. AB - BACKGROUND AND AIMS: Preoperative diagnosis of blunt diaphragamatic rupture is difficult and missed injuries can lead to severe late complications. The aim of this study was to assess the value and reliability of initial chest radiographs in diagnosing blunt diaphragmatic rupture. MATERIAL AND METHODS: A retrospective analysis of initial radiographs of 18 patients with blunt diaphragmatic rupture treated at two Greek Trauma Centers was performed. The chest radiograph was the primary diagnostic radiological tool in all of cases, and the findings were confirmed at operation in all cases. RESULTS: The preoperative diagnosis of blunt diaphragmatic rupture on the basis of chest radiographs was made in 16 out 18 patients (89%). The presence of air-containing viscera and an elevated nasogastric tube above the level of the left hemidiaphragm were the most specific signs. Although elevation of the hemidiaphragm was seen in all 18 cases, it is non-specific. A marked elevation of the right hemidiaphragm (more than 6 cm above the level of the left diaphragm), however, was a strong sign of right diaphragmatic rupture. CONCLUSIONS: In spite of the availability of newer diagnostic imaging techniques, the initial chest radiograph is very reliable in detecting most cases with blunt diaphragmatic rupture, and together with high index of suspicion and sound clinical assessment remain the cornerstone in diagnosing these challenging injuries. PMID- 10791639 TI - Computed tomography for staging of oesophageal cancer. AB - BACKGROUND AND AIMS: Computed tomography (CT) is still widely used in the staging of patients with carcinoma of the oesophagus. The aim of the study was to evaluate the diagnostic accuracy of CT in a series of patients with carcinoma of the oesophagus or the cardia. Results were compared with findings at surgery or autopsy. MATERIAL AND METHODS: 47 consecutive patients of whom 30 underwent operation while 17 patients were not candidates for surgery due to advanced disease. RESULTS: Demonstration of tumour invasion on CT in 30 patients who underwent operation had an accuracy of 63%, a nosographic sensitivity of 10% and a specificity of 90%. Diagnosis of metastases to abdominal lymph nodes had an accuracy of 57%, and a nosographic sensitivity and specificity of respectively 37% and 90%. The assessment of tumour invasion and metastases to lymph nodes in patients not candidates for surgery was expectedly more accurate (100 and 67%, respectively, in nine autopsies). CONCLUSION: CT may provide valuable information in pretherapeutic staging of oesophageal cancer by identifying patients with advanced disease, who are not candidates for surgery. In patients without signs of dissemination on CT additional information may be obtained from endoscopic ultrasonography and laparoscopy. PMID- 10791640 TI - Expandable metallic stents in the management of malignant oesophageal obstruction. AB - BACKGROUND: Malignant oesophageal obstruction with an advanced disease presents a difficult challenge. A new class of metal stents have been developed to overcome the limitations of existing treatment modalities. METHODS: We present our first 58 patients, who have been treated with self-expandable metallic stents, using sedation anaesthesia, with fluoroscopic and endoscopic control. Both kinds of stents, covered and uncovered, were applied. RESULTS: There was no procedure related mortality. The immediate relief of dysphagia was 98%. All four oesophageal fistulas were successfully sealed with covered stents. Due to stent migration, tumour overgrowth, or ingrowth, twelve (21%) of the patients needed re intervention. Restenting or laser therapies were used against recurrent dysphagia. CONCLUSION: The palliation of oesophageal malignant obstruction with metal stents is a rapid, effective, and relatively safe single treatment which can be employed as part of a multimodal treatment program. PMID- 10791641 TI - Video-assisted thoracoscopic surgery in the diagnosis of lung disease. The Cretan experience. AB - BACKGROUND: Video-assisted thoracoscopic surgery (VATS) has been recently utilised in the diagnosis and management of thoracic diseases. In this report we reviewed our VATS experience for biopsy of diffuse or localised lung diseases in 51 cases focusing on indications, operative procedures, complications or failures rates. PATIENTS AND METHODS: Over the last 5 years we performed 51 VATS procedures for diagnostic purposes in 32 men and 19 women. The specific indications for VATS were lung biopsy for undiagnosed diffuse or localised lung disease. In all patients the postoperative pain was controlled with the use of non-narcotic analgesics and was measured according the visual analogue scale (VAS). RESULTS: There was no operative mortality. Postoperative non-fatal complications were seen in 3 cases (6%). The overall median duration of chest tube drainage was 2 days and the mean postoperative stay 3 days. In the diffuse lung disease a tissue diagnosis was obtained in all the cases. Conversion to thoracotomy was needed in 1 case (2%), owing to extensive adhesions. All patients expressed a postoperative pain control effect of less than 50% of VAS. CONCLUSIONS: VATS should be considered as a safe and effective procedure, with low postoperative pain and morbidity. Should be recommended in patients who require a histological diagnosis of diffuse or localised lung diseases. PMID- 10791642 TI - Functional results of anterior levatorplasty and external sphincter plication for faecal incontinence. AB - BACKGROUND AND AIMS: The aim of this study was to assess results of anterior levatorplasty combined with external anal sphincter plication for faecal incontinence. MATERIAL AND METHODS: The study involved 44 female patients, 27 with idiopathic and 17 with traumatic faecal incontinence. All underwent anterior levatorplasty and external sphincter plication in our institution between 1986 and 1997. The patients were followed up clinically for a mean 12 (range 2-54) months and 39 (89%) patients also underwent pre-and postoperative anal manometry. RESULTS: In the idiopathic group 5 patients (19%) estimated that the result of the operation had been good and another 18 (67%) reported feeling better than before; the corresponding figures in the traumatic group were 4 (24%) and 10 (59%). The Wexner incontinence score decreased significantly after the operation in both groups. Nineteen patients (70%) in the idiopathic and 14 (82%) in the traumatic group showed improvement of one or more scores on the Kirwan scale. Three patients regained continence completely, one in the traumatic and two in the idiopathic group. There were no significant improvements in mean resting anal pressure or functional anal canal length in either group. Mean squeeze pressure improved significantly only in the traumatic group. CONCLUSIONS: Although the results of anterior levatorplasty combined with external sphincter plication are not perfect, the approach seems to be a valuable alternative in the treatment of patients with idiopathic or traumatic faecal incontinence. Complete continence is seldom achieved, but the grade of incontinence is reduced in the majority of patients. PMID- 10791643 TI - Conduction defects after coronary artery bypass grafting--a disappearing problem? AB - BACKGROUND AND AIMS: To evaluate the incidence of conduction defects (CDs) following coronary artery bypass grafting (CABG) in three different patient populations, to assess the etiologic factors associated with CDs, and to find out their effect on immediate postoperative outcome of the patient. MATERIAL AND METHODS: Three patient populations were prospectively studied: cohort A consisted of 180 CABG-patients operated between 1990-91, cohort B of 100 patients operated during the year 1993 and cohort C of 118 patients operated from April 1997 to June 1997. Cold crystalloid cardioplegia was used throughout the study years. In the first cohort A, two separate cavae were cannulated and clamped, venting through the right upper pulmonary vein was used, iced cold saline was used in pericardium, and cardioplegia was given until a myocardial temperature of 10-15 degrees of Celcius was attained. In the two later cohorts, two-stage venous cannula and aortic root venting were used and cardioplegia was given only until the activity of the myocardium stopped. Proximal anastomoses were performed after aortic declamping in cohort A, and during aortic occlusion in the two later cohorts. RESULTS: The incidence of permanent CDs in cohort A was 36%, in cohort B 5% and in cohort C 1%. Permanent atrioventricular (AV-) and left-sided blocks disappeared first. Left main coronary artery stenosis and low myocardial temperatures were associated with CDs. Patients with permanent CDs had more often low cardiac output after the operation, their values of cardiac enzymes were higher, and they had more often postoperative infarction than patients without CDs. CONCLUSIONS: The disappearance of all long lasting AV- and left-sided blocks simultaneously with decreasing plasma levels of cardiac enzymes is evidence that protection of both conduction tissue and myocardium had considerably improved in the two later cohorts. Giving cardioplegia in smaller amounts and more often at the same time when raising the general temperature during perfusion were the main reasons for the disappearance of postoperative CDs. PMID- 10791644 TI - Axillary-coronary artery bypass reconstruction as an alternative in coronary artery reoperations. AB - BACKGROUND AND AIMS: The increasing incidence of reoperations in coronary surgery associated with higher perioperative risks is a challenge for refinement of the surgical methods. The aim of the work is to prove the feasibility and satisfactory intermediate results of minimally invasive axillary-coronary artery bypass reconstruction in redo coronary surgery in case the left internal mammary artery had already been harvested. MATERIAL AND METHODS: Three patients (six months, two and six years after primary coronary artery bypass grafting) admitted for redo coronary surgery because of a recurrence of angina and proven malfunction of the left internal mammary artery-left anterior descending coronary artery anastomosis. An axillary-coronary venous graft was performed via left anterior small thoracotomy (LAST) on a beating heart in all three cases. RESULTS: Excellent patency of the graft was noted on control angiography within 9 days after the procedure together with good clinical improvement in midterm follow-up. CONCLUSION: Minimally invasive axillary-coronary artery bypass via LAST access in redo coronary surgery is a good alternative in cases where the left internal mammary artery cannot be used. PMID- 10791647 TI - Longitudinal split of peroneus brevis tendon. A report on two cases. AB - BACKGROUND AND AIMS: To describe the clinical findings and surgical treatment of peroneus brevis split. MATERIAL AND METHODS: Two cases of longitudinal split of the peroneus brevis tendon are reported. One of the patients was a healthy middle aged woman, who had fallen out of a car in a traffic accident and sprained her right ankle. Lateral ankle sprain was diagnosed and treated with a compression bandage. Lateral ankle pain persisted, however, with some swelling in the peroneal tendon region. MRI revealed a longitudinal partial rupture of the peroneus brevis tendon, which was treated surgically 12 months after the trauma. The second case was a 53-year-old woman, who had been suffering from rheumatoid arthritis for 2 years. Chronic pain and swelling in the peroneal tendon region were treated with 6 local corticosteroid injections without significant relief. Preoperative ultrasonography showed effusion of the peroneal tenosynovium, but the operation revealed a longitudinal split in the peroneus brevis tendon. RESULTS: In the first case, a single central peroneus brevis split was repaired with side-to-side suturation. After four weeks with a below-knee cast the patient was allowed to walk freely. At follow-up 12 months postoperatively, she was satisfied, although she still had some exertion pain in her ankle. In the second case, the torn fragment of the peroneus brevis tendon was excised and the ankle was mobilized early. Healing was complicated by a wound fistula, which was treated with antibiotics. Subluxation of the peroneus longus tendon necessitated a reoperation, which revealed a rerupture and a defect of the peroneus brevis tendon. The subluxation was repaired and the ruptured tendon ends were revised, followed by four weeks of below-knee cast immobilization, after which the patient was allowed to walk freely. The outcome was good. CONCLUSION: Peroneus brevis split easily goes unrecognised or misdiagnosed. It must be considered in patients with a history of single or recurrent ankle sprain or a chronic inflammatory disease. Lateral ankle pain, diffuse or local swelling in the peroneal tendon region, and a stable or instable ankle with no peroneal weakness are the main symptoms and findings. MRI is the most exact method for diagnosing tendon split. Surgical treatment usually gives good results. PMID- 10791645 TI - Treatment of subcapital femoral neck fractures with bioabsorbable or metallic screw fixation. A preliminary report. AB - BACKGROUND AND AIMS: Cancellous bone fractures and arthrodeses have been treated successfully with bioabsorbable polyglycolide and poly-L-lactide implants. In this study bioabsorbable poly-L-lactide lag screws and metallic screws were compared in the fixation of subcapital femoral neck fractures. PATIENTS AND METHODS: Forty patients with femoral neck fractures were treated by internal fixation using three bioabsorbable self-reinforced poly-L-lactide (SR-PLLA) lag screws of 6.3 mm in diameter and 38 patients using three metallic screws of 7 mm in diameter. In addition, one patient was operated on using two and one using four metallic screws. The patients did not differ in age, body weight or primary dislocation of the fracture. There were six males in the lactide group versus 14 in the metallic fixation group. There were two Garden Stage I, 27 Garden Stage II, nine Garden Stage III, and two Garden Stage IV fractures in both groups. RESULTS: In the Garden I and II fractures there were 5/29 redislocations after SR PLLA fixation and 8/29 after metallic fixation. In the Garden III fractures there were 4/9 and in the Garden IV fractures 2/2 redislocations in both groups. The ability to walk and the range of movement were better after bioabsorbable fixation. CONCLUSION: Self-reinforced poly-L-lactide lag screws can be used safely to fix subcapital femoral neck fractures in Garden Stage I and II fractures and in younger patients in Garden III fractures. PMID- 10791646 TI - Achilles tendon ruptures in South-East Finland between 1986-1996, with special reference to epidemiology, complications of surgery and hospital costs. AB - BACKGROUND AND AIMS: The incidence of achilles tendon (AT) ruptures is increasing. The aim of the present study was to evaluate annual incidence, aetiology, operative complications and direct hospital costs of AT ruptures. MATERIAL AND METHODS: A retrospective study of 93 consecutive patients operated on for AT rupture from January 1986 to December 1996 at Kuusankoski District Hospital (area with 92,500 inhabitants) was performed. During the observation period no patient with an AT rupture was treated conservatively. RESULTS: 95 AT ruptures were treated including one rerupture (1%) and one patient with two ruptures. There were 7 (7%) patients with an open AT rupture. The total annual incidence in the hospital area was 8.6 (+/- 4.3) and for closed AT ruptures 8.0 (+/- 3.8). The total incidence was 9.3 (+/- 4.6)/10(5) and for closed AT ruptures 8.6 (+/- 4.1)/10(5) inhabitants per year. Most of the injuries were sport related, the most frequent sport being volleyball. Patients operated for closed AT rupture had major surgical complications in 4.5% of the cases and the total complication rate was 11%. The average direct hospital costs per patient was USD 1375. CONCLUSIONS: The incidence of AT ruptures is increasing in South-East Finland. The rate of major surgical complication was low (4.5%) and comparable with earlier studies. PMID- 10791648 TI - Apoptosis in acute pancreatitis. AB - BACKGROUND: Apoptosis may be involved in the mechanism of acinar cell injury in acute experimental pancreatitis. AIM: This study was to investigate whether apoptosis also is involved in human acute pancreatitis. METHOD: A needle biopsy pancreatic specimen was obtained from a patient with acute oedematous pancreatitis. The specimen was stained with In Situ Cell Death Detection Kit. Similar specimen from a patient undergoing pancreatoduodenectomy for bile duct cancer served as a control. RESULTS: Extended acinar cell apoptosis was found in the pancreatitis specimen. No single apoptotic cell was found in the control pancreas. CONCLUSION: Apoptosis probably is involved not only in acute experimental pancreatitis but also in human acute oedematous pancreatitis. The induction and role of apoptosis in pancreatis is discussed based on literature. PMID- 10791649 TI - Abdominal vascular injuries. The trauma surgeon's challenge. PMID- 10791650 TI - The birth cohorts grow up: new opportunities for epidemiology. PMID- 10791651 TI - Overheated news of a 'medical breakthrough'. PMID- 10791652 TI - The changing prevalence of neural tube defects: a population-based study in the north of England, 1984-96. Northern Congenital Abnormality Survey Steering Group. AB - Using data from the Northern Congenital Abnormality Survey, we describe trends in the prevalence of neural tube defects (NTDs), maternal age-specific prevalence, outcome of pregnancy and the sensitivity of antenatal diagnosis of NTD-affected pregnancies in the Northern Region during 1984-96. This population-based sample consisted of 934 NTDs: there were 403 (43.1%) with anencephaly, 472 (50.5%) with spina bifida and 59 (6.3%) with encephalocele. The total prevalence at birth was 17.9 per 10,000 births and terminations (95% confidence interval [CI] = 16.7, 19.0) with a significant reduction over time (chi(2) for trend = 7.2; P = 0.007). The overall birth prevalence was 5.6 per 10,000 births [95% CI = 5.0, 6.3]; there was also a significant reduction in birth prevalence with time (chi(2) for trend = 68.3; P < 0.0001). Maternal age-specific prevalence rates decreased with increasing age. The proportion of NTD pregnancies terminated increased from 60.3% (325 cases) during 1984-90 to 78.6% (293 cases) during 1991-96, whereas the proportion of livebirths declined from 31.7% (171 cases) to 15.0% (56 cases) (P < 0.001). The sensitivity of antenatal diagnosis was consistently high for anencephaly (98%) and increased significantly for spina bifida from 60% during 1984-90 to 85% during 1991-96 (P < 0.05). Ascertainment of all cases of NTD in the Northern Region revealed a twofold reduction in birth prevalence between 1984 90 and 1991-96. This has resulted from improvements in the accuracy of antenatal detection of NTD-affected pregnancies with an increase in terminations of pregnancy. PMID- 10791653 TI - The Dutch 'Folic Acid Campaign'--have the goals been achieved? AB - Periconceptional folic acid use considerably reduces the risk of neural tube defects. The aim of this study was to measure the effect of the national and the local 'Folic Acid Campaign' on periconceptional folic acid use. Before (1995 survey) and 1 year after the campaign (1996 survey), the awareness and use of folic acid was measured among pregnant women in four regions of the Netherlands. To this end, pregnant women who visited the midwife, general practitioner or obstetrician for the first or second prenatal visit were asked to complete a questionnaire. The results showed that use of folic acid for any period around conception increased from 25.1% in 1995 to 53.5% in 1996. Appropriate use (4 weeks before until 8 weeks after conception) increased from 4.8% in 1995 to 21.0% in 1996. No additional effect of the local Folic Acid Campaign was found (adjusted odds ratio= 1.0; 95% confidence interval = 0.7, 1.4). It was possible to conclude that folic acid use at the recommended time increased considerably as a result of the national and the local Folic Acid Campaign, but the target (use in 46% of women wishing to conceive) was not achieved. New health education programmes are needed to increase further its use at the appropriate times. PMID- 10791654 TI - Maternal smoking affects fetal growth more in the male fetus. AB - This study investigated the association between maternal cigarette smoking and fetal growth, evaluated by longitudinal ultrasound examinations and by neonatal anthropometric measurements. The investigation was carried out in a healthy population of affluent Scandinavian women, parity 1 and 2, who were selected consecutively and prospectively, and with term, normal pregnancies. Three hundred and six non-smoking, 242 light-smoking and 308 heavy-smoking mothers and their newborns were examined. Ultrasound measurements were performed in pregnancy weeks 17, 25, 33 and 37. Biparietal diameter (BPD), mean abdominal diameter (MAD) and femur length were recorded. The negative effect on fetal growth from maternal smoking was found to affect the male fetus proportionally more than the female. Boys born to heavy-smoking mothers had a weight reduction of 8.2% and a lower fat accretion (as measured by subscapular skinfold) of 12%, whereas girls had a weight and fat reduction of 4.8% and 2% respectively. In boys (but not girls) born to smokers, head circumference was significantly smaller, also reflected by significantly smaller mean BPD measurements recorded from pregnancy week 18 onwards. The MAD measurements became successively more negatively affected in the second half of pregnancy in both males and females. A greater intrauterine growth velocity and a different hormonal milieu are suggested as possible explanations of the greater male susceptibility. PMID- 10791656 TI - Pre-eclampsia and premature labour among pregnant wowen with haematuria. AB - The authors conducted a retrospective cohort study of pregnant women to estimate the prevalence and associated risk of adverse pregnancy outcomes among pregnant women with microhaematuria. Between 1 January 1993 and 15 March 1993 at University of Washington Medical Center, 328 consecutive births were identified, and demographic data, medical history, laboratory data and pregnancy outcomes were abstracted from hospital charts. The presence or absence of haematuria was determined in 276/328 cases. Sixteen per cent (44/276) of the women were found to have haematuria. Those with haematuria were at increased risk of developing pre eclampsia (odds ratio [OR] = 9.5, 95% confidence interval [CI] 3.1, 28.2) and premature labour (OR = 3.8, 95% CI 1.5, 9.7). These associations persisted after controlling for age, race, and urinary tract infection (UTI) (pre-eclampsia OR=9.1, 95% CI 2.5, 33.7; premature labour OR = 4.2, 95% CI 1.2, 15.3). Infants of women with haematuria were at a non-significantly increased risk of low Apgar scores (OR = 2.8, 95% CI 0.7, 11.9) and low birthweight (OR=1.9, 95% CI 0.7, 4.7). In this observational study, microscopic haematuria was prevalent among pregnant women and was independently associated with an increased risk of adverse maternal complications. PMID- 10791655 TI - Stress and pregnancy among African-American women. AB - Research on stress tends to support an adverse effect on pregnancy outcomes, and suggests that the impact of these stressors is modified by social class and/or race. This study explicitly examined social factors such as experiences of discrimination, either racial or sexual, and neighbourhood crime as predictors of stress. We also examined cortisol and stress as predictors of blood pressure. A subsample of 94 African-American pregnant women, aged 18-39 years, who were enrolled in a longitudinal study of pregnancy and exposure to lead in the environment were used in this analysis. The women were patients at an obstetrics clinic at Magee Women's Hospital in Pittsburgh, PA, USA. Younger age, higher income, lower education and experiences of discrimination, both racial and sexual, were related to greater perceived stress; however, life events were not related to perceived stress. Higher income and urinary cortisol adjusted for creatinine were related to systolic blood pressure after the 36th week. As a body of evidence suggests that stress can have deleterious effects in both pregnant and non-pregnant women, future research should examine these forms of discrimination, especially racial discrimination, as a possible reason for the disparity in adverse pregnancy outcomes between African-American and white women. PMID- 10791657 TI - Maternal pre-eclampsia/eclampsia and the risk of sudden infant death syndrome in offspring. AB - To determine whether maternal exposure to pre-eclampsia/eclampsia during pregnancy increases the risk of sudden infant death syndrome (SIDS) in offspring, we conducted a population-based case-control study using the California linked birth and death certificate data. All infants who died of SIDS (ICD-9 code 798.0) during 1989-91 were identified as cases. More than 96% of the identified SIDS cases were diagnosed through autopsy. Ten controls who did not die from SIDS were randomly selected for each case from the birth certificate matched to the case on the year of birth. Among 2,029 cases and 21,037 controls included in the final analysis, mothers of 49 cases (2.4%) and 406 controls (1.9%) had a diagnosis of either pre-eclampsia or eclampsia noted on the birth certificate. After adjustment for maternal age, prenatal smoking, race/ethnicity, parity, maternal education, gestational age at the initial visit for prenatal care, infant year of birth and infant sex, maternal pre-eclampsia/ eclampsia during pregnancy was associated with a 50% increased risk of SIDS in the offspring (odds ratio = 1.5, 95% confidence interval 1.1, 2.0). Potential under-reporting of pre eclampsia/eclampsia on the birth certificates was likely to be non-differential and is unlikely to explain the finding. Fetal hypoxia resulting from pre eclampsia/ eclampsia or immunological aetiology affecting the risk of both pre eclampsia/eclampsia and SIDS may explain the finding. PMID- 10791658 TI - Maternal age and preterm births in a black population. AB - Babies born to teenagers aged 15-19 years have a substantial risk of dying within the first year of life. Although associated socio-demographic factors may account for an increase in the risk of adverse reproductive outcomes for teenagers, there is a concern that young maternal age may also be a biological risk factor. We examined the effects of maternal age of primiparous black women on the incidence of preterm births using data from 6,072 black women delivering between 1989 and 1995 at an urban perinatal network of 17 hospitals and health centres serving residents in a well-defined geographical area. Maternal age was grouped as: < or = 15, 16-17, 18-19, 20-24, 25-29 or > or = 30 years age groups. The 20-24 age group with the highest number of births and lowest preterm rate was used as the reference age. Preterm birth was defined as delivery < 37 completed weeks of gestation. Of the 6,072 infants born to the cohort, 1,170 (19.3%) were preterm. The unadjusted odds for a preterm birth for the < or = 15-year-olds (odds ratio [OR] = 0.97; 95% confidence interval [CI], 0.69,1.36), for the 16- to 17-year olds (OR=1.21; CI=0.94, 1.57) and for the 18- to 19-year olds (OR=1.15, CI = 0.92, 1.43) were not significantly different from that for the reference group. The risk for the 25-to 29-year-old mothers was 1.26 times [CI = 1.05, 1.50] and for the > 30-year-old mothers 1.28 times [CI = 1.07, 1.52] that for the reference group. Adjustments using logistic regression analysis for the effects of maternal smoking, drug abuse during pregnancy, insurance status, having prenatal care and median family income from census tract of residence did not result in a significantly increased risk for preterm birth or low birthweight for the teenage groups compared with the reference group. We conclude that primiparous teenage black mothers do not have an inherent biologically increased risk for preterm births. PMID- 10791659 TI - Racial differences in temporal changes in newborn viability and survival by gestational age. AB - This study examines trends in the rates of very preterm, moderately preterm and gestational age-specific neonatal mortality, and in the gestational age limit of viability in South Carolina (SC) from 1975 to 1994. We also investigate whether trends were similar between African-Americans and Whites. We hypothesised that disproportionate reductions in gestational age-specific mortality, rather than any major changes in the gestational age distributions of either race group, underlie any increasing racial disparity in overall mortality rates. During 1975 94, single livebirths, who were born to mothers resident in SC and were either White or African-American based on recorded maternal race, were selected for the investigation. We define the gestational age limit of viability as the gestational age at which > or = 50% of infants in the population died within 28 days of life. Although preterm percentages have not improved, there was a marked decline in neonatal mortality. Gestational age-specific neonatal mortality decreased for both race groups, although there were greater reductions for White preterm infants. By the end of the study period, the African-American neonatal mortality rate was 2.3 times that of Whites and the gestational age at which 50% of newborns died within 28 days of life was 24.5 weeks for Whites and 23.9 weeks for African-Americans. The ongoing decline in neonatal mortality continues to be mainly due to reductions in gestational age-specific neonatal mortality, probably related to high-risk obstetric and neonatal care. Technological developments in these areas may have differentially benefited Whites, resulting in an increasing racial disparity in neonatal mortality rates. Preterm African-American infants no longer have a marked survival advantage over White infants, even at the gestational age limit of viability. PMID- 10791660 TI - Birthweight and infant mortality in Bulgaria's transition crisis. AB - Between 1988 and 1991, years of political and economic crisis, Bulgaria reported a 25% increase in infant mortality. From 1991 to 1995, the rate then dropped slightly. Analysis of detailed unpublished vital statistics shows that the reported increase could not have been a result of more complete reporting of early neonatal mortality for very low-weight births. Old reporting practices continue unchanged. Rather, these statistics reveal an increase in the proportion of low-weight births, combined with rising death rates, particularly for low weight births in the postneonatal period, but also for normal-weight babies. PMID- 10791661 TI - Survival of infants born with Down's syndrome: 1980-96. AB - The improved life expectancy of people with Down's syndrome as a result of the greater availability of surgery and advances in medical care has been widely documented. However, there has been no evaluation of survival in the Australian Down's syndrome population since the 1980s. This study aimed to evaluate the changes in survival from birth in cases of Down's syndrome notified to the Birth Defects Registry in Western Australia. Babies born with Down's syndrome between 1980 and 1996 (inclusive) and registered with the Birth Defects Registry were studied. Survival status was obtained in several ways. Cases were stratified into three cohorts for comparison. Survival curves were constructed using the methods of Kaplan and Meier. For infants born during 1980-96, survival to 1 year is now > 91%, and 85% can expect to survive until the age of 10 years. Although survival in those with heart disease showed improvement over the period studied, overall this was still a strong predictor of mortality. Survival in Aboriginal children with Down's syndrome was significantly poorer than in non-Aboriginal children, mirroring the pattern in the general population. Mortality was greater in females and in those with a low birthweight. There was no statistically significant difference in the survival between those born in metropolitan and in rural areas. There has been a considerable improvement in survival of infants born with Down's syndrome in Western Australia. This improvement is similar to findings in recent international studies. The difference in survival between Aboriginal and non Aboriginal children is particularly disturbing. These findings are useful for both clinicians and families who need to plan for the long-term care of these children. PMID- 10791662 TI - Methods to improve the reliability of histopathological diagnoses in the placenta. AB - The interobserver reliability of histopathological features in the placenta was examined. Two pathologists independently reviewed slides from 250 placentas. The pathologists were given a morphological description of the placenta, but were blinded to clinical status, gestational age and original diagnoses. A protocol for diagnosis and grading of features was first developed and pilot-tested. Definitions and criteria were refined and elaborated. A range of features was examined including inflammatory lesions, features indicative of reduced uterine blood flow and other miscellaneous histopathological changes. Weighted kappa coefficients were calculated. The effect of multiple features on reliability was examined by stratifying on the presence of a second feature and calculating stratum-specific kappa coefficients. Results indicated good to excellent agreement for diagnoses of chorioamnionitis, cord vasculitis, funisitis and villitis (kappa(w) range 0.70-0.83). Agreement between observers was more variable for the diagnosis of reduced uterine blood flow states. Excellent agreement was observed for the diagnosis of meconium staining of the placenta (kappaw = 0.79). In general, lower levels of agreement were observed for features in the presence of a second feature. Reproducible measures are a prerequisite to using placental histopathology for diagnostic and prognostic information. This study demonstrated reliable placental diagnoses can be achieved through a standardised protocol. PMID- 10791663 TI - Evaluation of the Dinamap 8100 and Omron M1 blood pressure monitors for use in children. AB - The aim of this study was to evaluate the Dinamap 8100 and the Omron M1 (the test devices) against standard criteria for use in children in a fieldwork setting. Device calibration and validation were carried out in accordance with the British Hypertension Society protocol for special groups such as children. A total of 102 children, recruited from infant and junior schools in Southampton, had six sequential measurements made of their blood pressure-four measurements with a mercury sphygmomanometer and two with one of two test devices, 55 children with the Dinamap 8100 and 47 with the Omron M1. Systolic and diastolic readings with the Dinamap 8100 were on average 11 mmHg higher (95% confidence interval [CI] +9, +12 mmHg) and 3 mmHg lower (95% CI -5, -1 mmHg), respectively, than measurements with the mercury sphygmomanometer, overestimating systolic pressures and underestimating diastolic pressures across the whole range observed. The Omron M1 gave readings lower by 1 mmHg on average for systolic pressures and 2 mmHg for diastolic pressures compared with the sphygmomanometer (95% CIs -4, +1 mmHg and 5, +1 mmHg respectively), specifically overestimating higher pressures and underestimating lower pressures. According to the criteria of the British Hypertension Society, neither the Dinamap 8100 nor the Omron M1 can be recommended for use in children in clinical situations in which accuracy of the absolute measurement is required. In epidemiological surveys, in which differences in blood pressure between groups of people are more important than absolute levels, it may be more appropriate to use these devices. Of the two, its more consistent performance supports the Dinamap 8100 as the instrument of choice in such studies of children. PMID- 10791664 TI - Well-leg compartment pressures during hemilithotomy position for fracture fixation. AB - OBJECTIVE: To evaluate the relationship between the well-leg compartment pressures and time during hemilithotomy position for fracture fixation. DESIGN: Prospective. SETTING: Level 1 trauma center. PATIENTS/PARTICIPANTS: Ten patients who underwent intramedullary nailing of a fractured femur in the hemilithotomy position (with a well-leg holder). INTERVENTION: Continuous pressure monitoring was achieved with in-dwelling slit catheters inserted into the calf compartments of the well leg. Baseline measurements were obtained in the supine position. After the leg was placed in the hemilithotomy position, compartment pressures were monitored throughout surgery. MAIN OUTCOME MEASUREMENTS: Calf compartment pressures at baseline, during hemilithotomy position, and post-hemilithotomy were compared. The association between body mass index and compartment pressure was analyzed. RESULTS: A consistent pattern was observed between compartment pressures and time. The curve was that of a step function in which the pressure increased as soon as the leg was placed in the well-leg holder and remained elevated until the leg was taken down. The pressure jumped from a baseline of 9.2 to 27.3 millimeters of mercury (mm Hg) (p<0.0001). While in the hemilithotomy position, the leg pressure trended slightly upward. Once the leg was taken down, the pressure immediately returned to a near-baseline level of 8.1 mm Hg (p<0.0001). A significant correlation was also found between the body mass index and leg pressure (R2 = 0.713; F = 0.002). CONCLUSIONS: The use of the well-leg holder to maintain hemilithotomy position increases the calf compartment pressures dramatically and significantly. Therefore, we recommend avoiding this position for fracture fixation in at-risk patients. PMID- 10791665 TI - Randomized prospective study of humeral shaft fracture fixation: intramedullary nails versus plates. AB - OBJECTIVES: To compare the clinical and radiographic results for locked intramedullary (IM) nails and plates used in the treatment of humeral diaphyseal fractures. DESIGN: Prospective randomization by sealed-envelope technique of eighty-four patients into two study groups: those treated by intramedullary nailing (IMN group; n = 38) and those treated by compression plating (PLT group; n = 46). SETTING: Patients admitted consecutively to a university-affiliated Level I trauma center. PATIENT/PARTICIPANTS: All skeletally mature patients admitted to Harborview Medical Center with acute humeral shaft fractures requiring surgical stabilization. Fractures of the diaphysis were defined as being at least three centimeters distal to the surgical neck and at least five centimeters proximal to the olecranon fossa. INTERVENTION: Treatment with locking antegrade intramedullary humeral nails (Russell-Taylor design [Smith and Nephew Richards]) or with 4.5-millimeter dynamic compression and limited contact dynamic compression plates (AO design [Synthes]). MAIN OUTCOME MEASUREMENTS: Clinical outcome measurements included fracture healing, radial nerve recovery, infection, and elbow and shoulder discomfort. Radiographic measurements included fracture alignment, time to healing, delayed union, and nonunion. RESULTS: Follow-up averaged thirteen months. Forty-two fractures (93 percent) in the PLT group were healed by sixteen weeks versus thirty-three fractures (87 percent) in the IMN group (p = 0.70). Shoulder pain and a decrement in shoulder range of motion (ROM) were significant associations with IMN (p = 0.007 for both variables) but not with PLT. A decrement in elbow ROM was significantly associated with PLT (p = 0.03), especially for fractures of the distal third of the diaphysis, whereas elbow pain was not (p = 0.123). The sum of other complications demonstrated nearly equal prevalence for both treatment groups. CONCLUSIONS: For patients requiring surgical treatment of a humeral shaft fracture, intramedullary nailing and compression plating both provide predictable methods for achieving fracture stabilization and ultimate healing. PMID- 10791666 TI - Somatosensory evoked potential monitoring during closed humeral nailing: a preliminary report. AB - OBJECTIVES: To assess the role of intraoperative somatosensory evoked potential (SSEP) monitoring of the radial and median nerves in preventing iatrogenic nerve injury during closed, locked intramedullary (IM) nailing of the humerus. DESIGN: Prospective clinical study. SETTING: Pacific Northwest Level One trauma center and Southern California military medical center. PATIENTS: Thirteen patients with indications for surgical stabilization of fractures of the humeral diaphysis and either unknown neurologic status of the affected limb or anticipated difficult reduction maneuvers due to fracture complexity or displacement. INTERVENTION: Closed, antegrade or retrograde locked IM nailing of the humerus was attempted while intraoperative monitoring of the radial and median nerves with SSEP was performed. MAIN OUTCOME MEASUREMENTS: Intraoperative radial and median nerve SSEP changes during closed fracture manipulation, guide rod insertion, reaming, and humeral nail placement. RESULTS: Baseline recordings were obtained in twelve of thirteen patients for both the radial and median nerves. An absence of radial nerve signal in one patient with a closed head injury prompted an open procedure, revealing entrapment of the radial nerve in the fracture. Intraoperative SSEP changes were observed in two of the twelve remaining patients during fracture manipulation and distal interlocking. The signal amplitude returned after discontinuation of manipulation and traction, and alteration of the interlocking maneuver. No neurologic deficits were noted in these two patients. CONCLUSIONS: Intraoperative radial nerve SSEP monitoring appears to reliably reflect the status of the radial nerve in those patients with a humerus fracture. In three of eleven patients, intraoperative signal changes prompted a change in surgical plan. In no patient did there appear to be evidence of iatrogenic nerve injury. PMID- 10791667 TI - Fracture of the radial head with associated elbow dislocation: results of treatment using a floating radial head prosthesis. AB - OBJECTIVES: To assess elbow function, complications, and problems of radial head fractures associated with elbow dislocation receiving surgical treatment with a floating prosthesis. DESIGN: Prospective clinical study. SETTING: University Hospital, Orthopaedic Department, Sart Tilman, Liege, Belgium. PATIENTS: Eleven consecutive adult patients were treated with a floating prosthesis for acute radial head fractures associated with elbow dislocation from January 1994 to September 1996. INTERVENTION: The floating radial head prosthesis (Tornier SA, Saint-Ismier, France) was used in all our patients. The implant is in two parts: a radial head made of high-density polyethylene enclosed in a cobalt-chrome cup, which articulates in a semiconstrained manner with the spherical end of a cemented intramedullary stem. The implants were inserted within the first week following the injury (range 2 to 7 days). Three cases also required internal fixation of the coronoid process of the ulna; in one case plate fixation of an olecranon fracture was also performed. MAIN OUTCOME MEASUREMENTS: Patients were assessed by physical examination, a functional rating index (Morrey et al.), and radiographs. The parameters evaluated were motion, stability, pain, and grip strength. Potential complications such as infection, prosthetic failure, or dislocation were investigated. RESULTS: The minimum follow-up time was two years (mean 32 months, range 24 to 56 months). Four patients were considered to have excellent results, four patients were considered to have good results, two patients had fair results, and one patient had a poor result. There were no cases of infection, prosthetic failure, or dislocation. No patient required prosthetic revision. CONCLUSION: The basic principle of maintaining anatomic and physiologic relationships applies when deciding on treatment for radial head fractures with associated elbow dislocation. The loss of lateral osseous support will render the elbow grossly unstable. We believe that a floating prosthesis may be indicated in Mason Type III radial head fractures associated with elbow dislocation, especially in the presence of associated destabilizing fractures. Well-controlled comparative randomized studies will be needed to delineate the optimal treatment for a given situation. PMID- 10791668 TI - Success of exchange reamed intramedullary nailing for femoral shaft nonunion or delayed union. AB - OBJECTIVES: To investigate the success of exchange reamed femoral nailing in the treatment of femoral nonunion after intramedullary (IM) nailing, and to analyze factors that may contribute to failure of exchange reamed femoral nailing. DESIGN: Retrospective consecutive clinical series. SETTING: Level I trauma center and tertiary university hospital. PATIENTS: Twenty-three patients were identified whose radiographs failed to show progression of healing for four months after treatment with a reamed IM femoral nail. Nineteen patients had undergone primary IM nailing of an acute femoral shaft fracture, one patient had been converted to an IM nail after initially being treated in an external fixator, and three patients had previously undergone an unsuccessful exchange reamed nailing. INTERVENTION: All patients were treated by exchange reamed femoral nailing. The diameter of the new nail was one to three millimeters larger than that of the previous nail (the majority were two millimeters larger). The intramedullary canal was overreamed by one millimeter more than the diameter of the nail. Most of the nails were statically locked, and care was taken to avoid distraction of the nonunion site by reverse impaction after distal interlocking was performed or by applying compression with a femoral distractor. MAIN OUTCOME MEASUREMENTS: Radiographic evaluation of union was determined by the presence of healing on at least three of four cortices. Factors reviewed included the patient's age, smoking history, mechanism of injury, associated injuries, whether the initial fracture was open or closed, the pattern and location of the fracture, the type of nonunion, the increase in nail diameter, whether the nail was dynamically or statically locked, and the results of any intraoperative cultures. RESULTS: Tobacco use was found to have a detrimental impact on the success of exchange reamed nailing. All eight of the nonsmokers healed after exchange reamed nailing, whereas only ten of the fifteen smokers (66.7 percent) healed after exchange reamed nailing. Overall, exchange reamed femoral nailing was successful in eighteen cases (78.3 percent). Three patients achieved union with additional procedures. Intramedullary cultures were positive in five cases; all of these achieved successful union. CONCLUSIONS: Exchange reamed nailing remains the treatment of choice for most femoral diaphyseal nonunions. Exchange reamed IM nailing has low morbidity, may obviate the need for additional bone grafting, and allows full weight-bearing and active rehabilitation. Tobacco use appears to have an adverse effect on nonunion healing after exchange reamed femoral nailing. PMID- 10791669 TI - Comparison of magnetic resonance imaging with operative findings in acute traumatic dislocations of the adult knee. AB - OBJECTIVES: To compare magnetic resonance imaging (MRI) with examination under anesthesia and with surgical findings in evaluating soft tissue injuries in acute traumatic knee dislocations in adults. DESIGN: Retrospective analysis. SETTING: Level I trauma center. PATIENTS: For a single surgeon, all patients who underwent MRI before surgical treatment for knee dislocations (ten individuals). INTERVENTION: Incompetent ligaments were repaired or reconstructed. MAIN OUTCOME MEASUREMENTS: MRI of knee dislocations was compared with clinical examination under anesthesia and with intraoperative findings at arthrotomy in ten cases. Pertinent positive and negative findings were recorded, and accuracy, sensitivity, specificity, and positive and negative predictive values were calculated. RESULTS: Two mid-grade sprains of the anterior cruciate ligament were erroneously read as complete tears. One rupture or avulsion of each the biceps tendon, the lateral collateral ligament, and the posterolateral and posteromedial corners were considered intact on MRI. The MRI studies erroneously identified tears of the lateral collateral ligament and medial meniscus in one case each. Otherwise, the study was highly accurate. CONCLUSIONS: MRI is useful for defining the presence of ligamentous injuries in knee dislocations; however, clinical examination under anesthesia is more accurate. PMID- 10791670 TI - A prospective, randomized study of intramedullary nails inserted with and without reaming for the treatment of open and closed fractures of the tibial shaft. AB - OBJECTIVES: To determine if there are differences in healing, complications, or number of procedures required to obtain union among open and closed tibia fractures treated with intramedullary (IM) nails inserted with and without reaming. DESIGN: Prospective, surgeon-randomized comparative study. SETTING: Level One trauma center. PATIENTS: Ninety-four consecutive patients with unstable closed and open (excluding Gustilo Grade IIIB and IIIC) fractures of the tibial shaft treated with IM nail insertion between November 1, 1994, and June 30, 1997. INTERVENTION: Interlocked IM nail insertion with and without medullary canal reaming. MAIN OUTCOME MEASURES: Time to union, type and incidence of complications, and number of secondary procedures performed to obtain union. RESULTS: For open fractures, there were no significant differences in the time to union or number of additional procedures performed to obtain union in patients with reamed nail insertion compared with those without reamed insertion. A higher percentage of closed fractures were healed at four months after reamed nail insertion compared with unreamed insertion (p = 0.040), but there was not a difference at six and twelve months. More secondary procedures were needed to obtain union after unreamed nail insertion for the treatment of closed tibia fractures, but the difference was not statistically significant given the limited power of our study (p = 0.155). Broken screws were seen only in patients treated with smaller-diameter nails inserted without reaming, and the majority occurred in patients who were noncompliant with weight-bearing restrictions. There were no differences in rates of infection or compartment syndrome. CONCLUSION: Our findings support the use of reamed insertion of IM nails for the treatment of closed tibia fractures, which led to earlier time to union without increased complications. In addition, canal reaming did not increase the risk of complications in open tibia fractures. PMID- 10791671 TI - Tibial shaft fractures with an associated infrapopliteal arterial injury: a survey of vascular surgeons opinions on the need for vascular repair. AB - OBJECTIVE: To determine current practice for tibial arterial revascularization in trauma patients with tibial shaft fractures associated with infrapopliteal arterial injuries. DESIGN: Nationwide survey of board-certified vascular surgeons, proportionally stratified by geographic region. METHODS: We surveyed a random sample (probability sample stratified by geographic region) of 200 members of the 729 active members of the Society for Clinical Vascular Surgery. We designed a twenty-one-item questionnaire to elicit opinions on the need for vascular repair for a variety of clinical scenarios. RESULTS: Completed questionnaires were returned by fifty-one vascular surgeons. For the eight isolated and combined infra-popliteal arterial injuries we studied, agreement between vascular surgeons as to the need for arterial repair was better for scenarios with clinical evidence of limb ischemia than for those without clinical evidence of limb ischemia. For injuries with clinical evidence of limb ischemia, excellent agreement (90 percent or more of respondents agreeing) was seen for five of the eight injuries, good agreement (80 to 89 percent of respondents agreeing) was seen for two injuries, and poor agreement (less than 70 percent of respondents agreeing) was seen for one injury. For injuries without clinical evidence of limb ischemia, excellent agreement was seen for one injury, good agreement was seen for two injuries, fair agreement (70 to 79 percent of respondents agreeing) was seen for three injuries, and poor agreement was seen for two injuries. CONCLUSIONS: A review of the literature and results of our study suggest that no standardized protocol exists in the current practice of revascularization of infrapopliteal arterial injuries with concomitant tibial shaft fractures; disagreement among vascular surgeons was particularly common for cases where a vessel was known to be injured but there was no clinical evidence of limb ischemia. Our study highlights the need for randomized prospective studies so that standardized protocols can be developed for these serious injuries. Because of the relatively small numbers of this type of injury and the wide variety of injury patterns seen, a study such as this would best be designed as a multi-center study. PMID- 10791673 TI - Evaluation of standard surgical preparation performed on superficial dermal abrasions. AB - OBJECTIVES: To determine the difference, if any, between the reduction of bacteria on contaminated normal skin and contaminated superficially abraded skin following standard surgical preparations at clinically relevant time points after injury. DESIGN: Prospective animal study. SETTING: Laboratory. SUBJECTS: Thirty two New Zealand white rabbits. INTERVENTION: Two sites, two by two centimeters, one abraded and one nonabraded (control), were studied on each rabbit. Both were inoculated with encapsulated Staphylococcus aureus strain Wood 46. Four six millimeter punch biopsies were obtained after inoculation, immediately before surgical scrub, and five minutes and then two hours after completion of the surgical scrub. The rabbits were divided into four cohort groups with surgical scrubs performed at six, twelve, twenty-four, and forty-eight hours after inoculation. Bacterial counts were determined. MAIN OUTCOME MEASUREMENTS: Numbers of bacteria on surgical sites. RESULTS: Before surgical preparation, the amount of bacteria on the normal skin (control sites) dropped significantly (p<0.02) except in the six-hour group (p<0.20). At the abraded skin sites, the bacteria flourished. The surgical scrub dropped bacterial counts at both the abraded and nonabraded skin sites significantly (p<0.05) except for the abraded site in the twenty-four-hour group (p<0.08). However in the twelve-, twenty-four-, and forty eight-hour groups, the bacterial counts (colony-forming units) were still markedly elevated (>1x10(5) at abraded sites) when compared with the nonabraded skin sites (p<0.008) at the respective time intervals. Only at the six-hour interval were the bacterial counts reduced similarly at both the abraded and nonabraded skin sites. CONCLUSIONS: In a rabbit model the standard surgical preparation using povidone-iodine at six hours after inoculation is effective in reducing the bacterial count on abraded skin to that of surgically prepared nonabraded skin. Beyond that time, the standard surgical preparation is ineffective in reducing counts to those of nonabraded skin at similar time intervals. PMID- 10791672 TI - On the behavior of closed tibial fractures: clinical/radiological correlations. AB - OBJECTIVE: To review the treatment of closed diaphyseal tibial fractures with functional braces and to identify patient and fracture characteristics, as well as possible correlations, that could be used to determine final outcomes. DESIGN: A comprehensive review of data obtained from 1,000 diaphyseal tibial fractures. SETTING: Major university teaching hospital. INTERVENTION: Identification of patient and fracture characteristics, including type and level of fracture, initial shortening and angulation, and severity of injury, to be correlated with final shortening and angulation and speed of healing. MAIN OUTCOME MEASUREMENTS: Degree of shortening and angulation; speed of healing. RESULTS: Neither the level nor the type of tibial fracture significantly influenced healing time, although there was a considerably higher probability of delayed union in fractures produced from vehicular accidents, as well as in comminuted and segmental fractures. Any delay in application of the functional brace resulted in slower healing. Maximum shortening of the fractures occurred at the time of the initial injury, with no additional shortening experienced after the introduction of graduated weight-bearing ambulation. Compared with a mean initial shortening of 4.25 millimeters, the overall final shortening of fractures was 4.28 millimeters. CONCLUSIONS: Correlations exist between patient and fracture characteristics that influence the final outcome for closed diaphyseal fractures treated with functional braces. PMID- 10791674 TI - Retained broken wound drains: a preventable complication. AB - A simple technique is described to avoid inadvertent suturing of a drain during wound closure. Drains that have been inadvertently sutured frequently break during removal, leaving a retained fragment in the wound. Use of the technique described herein will prevent this potentially avoidable complication. PMID- 10791675 TI - Gerdy's tubercle osteotomy for the treatment of coronal fractures of the lateral femoral condyle. AB - Coronal fractures of the femoral condyle (Hoffa fractures) are intraarticular fractures that are commonly treated surgically by open reduction and internal fixation. Surgical fixation is demanding because anatomic reduction is mandatory and adequate exposure is often difficult. Herein we describe a new technique that permits excellent visualization and fixation of lateral Hoffa fractures based on osteotomy of Gerdy's tubercle and reflection of the attached iliotibial band. PMID- 10791676 TI - Bilateral peroneal nerve injuries in a patient with bilateral femur fractures: a case report. AB - The second reported case in the current literature of peroneal nerve palsy in bilateral femur fractures is described. This is the first case report of bilateral nerve palsies occurring in bilateral femoral fractures and the first report of bilateral peroneal nerve palsy associated with bilateral skeletal traction. PMID- 10791677 TI - Nonunion in a fracture of the proximal phalanx of the thumb. AB - We describe the management of atrophic nonunion in a fracture of the proximal phalanx of the thumb in a patient who presented one and a half years after the injury. By using threaded external fixators applied across the fracture nonunion, stability for active mobilization of the metacarpophalangeal joint was achieved. The gradual distraction resulted in correction of the deformity and the shortening, as well as creation of an adequate space between the bone ends. Bone grafting and compression of the fracture with the same fixators followed. Consolidation of the fracture occurred within five months. Follow-up examination two and a half years postoperatively revealed complete remodeling of the fracture and excellent function of the hand. PMID- 10791678 TI - Intraoperative fluoroscopy. PMID- 10791679 TI - Plate osteosynthesis. PMID- 10791680 TI - Multiple equivalent current dipole source localization of visual event-related potentials during oddball paradigm with motor response. AB - Event-related potentials (ERPs) during a visual oddball paradigm with button pressing responses were recorded in 12 right-handed subjects from 32 scalp electrodes. The single equivalent current dipole (ECD) of the target C1 (weak occipito-parietal negativity from 30-80 ms) was consistently located at the primary visual cortex. From the 4-ECD localization of the target P1/N1 (temporally coincident frontal positivity and occipito-temporal negativity), it was suggested that this complex reflected activities from distributed sources along both dorsal occipito-parietal and ventral occipito-temporal areas. The stable multiple ECD solutions for the target P3b were chosen as those including the left primary motor and/or sensorimotor dipole and satisfying goodness-of-fit (GOF) of more than 98% and confidence limit (CL) of less than 1 mm. The obtained frontal dipoles were discussed in terms of visual working memory and sustained attention in reference to the previous PET, fMRI and MEG studies. The distributed multiple ECDs may suggest that P3 should be interpreted as being the embodiment of the cortico-limbic-thalamic network which involves Halgren and Marinkovic's emotional and behavioral model and Mesulam's attentional circuit. PMID- 10791681 TI - Mu and beta rhythm topographies during motor imagery and actual movements. AB - People can learn to control the 8-12 Hz mu rhythm and/or the 18-25 Hz beta rhythm in the EEG recorded over sensorimotor cortex and use it to control a cursor on a video screen. Subjects often report using motor imagery to control cursor movement, particularly early in training. We compared in untrained subjects the EEG topographies associated with actual hand movement to those associated with imagined hand movement. Sixty-four EEG channels were recorded while each of 33 adults moved left- or right-hand or imagined doing so. Frequency-specific differences between movement or imagery and rest, and between right- and left hand movement or imagery, were evaluated by scalp topographies of voltage and r spectra, and principal component analysis. Both movement and imagery were associated with mu and beta rhythm desynchronization. The mu topographies showed bilateral foci of desynchronization over sensorimotor cortices, while the beta topographies showed peak desynchronization over the vertex. Both mu and beta rhythm left/right differences showed bilateral central foci that were stronger on the right side. The independence of mu and beta rhythms was demonstrated by differences for movement and imagery for the subjects as a group and by principal components analysis. The results indicated that the effects of imagery were not simply an attenuated version of the effects of movement. They supply evidence that motor imagery could play an important role in EEG-based communication, and suggest that mu and beta rhythms might provide independent control signals. PMID- 10791682 TI - Stationarity and redundancy of multichannel EEG data recorded during generalized tonic-clonic seizures. AB - To improve our understanding of the physiology of generalized tonic-clonic (GTC) seizures, we have investigated the stationarity and redundancy of 21-electrode EEG data recorded from ten patients during GTC seizures elicited by electroconvulsive therapy (ECT). Stationarity was examined by calculating probability density functions (pdfs) and power spectra over small equal-length non-overlapping time windows and then by studying, visually and quantitatively, the evolution of these quantities over the duration of the seizures. Our analysis shows that some seizures had no demonstrable stationarity, that most seizures had time intervals of at least a few seconds that were statistically stationary by several criteria, and that, in some seizures, there were leads which were delayed in manifesting the statistical changes associated with seizure onset evident in other leads. The redundancy analysis demonstrated for the first time posterior-to anterior time delays in the mid-ictal region of GTC seizures. The implications of these findings are discussed for the analysis of GTC seizure EEG data, for the physiology of GTC seizures, and for ECT research. PMID- 10791683 TI - Two dimensional inverse imaging (2DII) of current sources in magnetoencephalography. AB - A new magnetoencephalographic (MEG) technique for imaging the cortical distribution of neuronal activity is described. An iterative algorithm is employed, which successively alters an initial estimate of cortical source structure until it corresponds to the measured magnetic field data. In this new technique, the continuum of electrical activity across the cortical surface is modeled as a dense grid of thousands of single equivalent current dipoles. MEG imaging of both compact and extended sources is facilitated by a wavelet-like transformation of the source space into a sequence of successively smaller composite source structures. Two of these composite source structures are combined during each iterative step to generate an improved estimate of the cortical source structure. Thus, inversion of the complete gain matrix corresponding to thousands of cortical sources is not performed. The technique requires only moderate PC based resources even for very large source grids. In contrast to minimum norm MEG imaging methods, this new algorithm is insensitive to random noise in the data. If available, prior knowledge of source structure from other imaging techniques, such as PET, MRI and fMRI, is easily incorporated as additional constraints on the source structure solution. Source images solutions corresponding to simulated data are presented. In addition, the technique is applied to source imaging of real MEG data incorporating cortical structure from volumetric MRI data. These results demonstrate the capability of our new technique for imaging combinations of compact and extended source structures. PMID- 10791684 TI - Estimating scalp MEG from whole-head MEG measurements. AB - Studies based on whole-head MEG recordings are providing more and more impressive results. In such recordings, the MEG sensors are several centimeters away from the scalp and the positions of the MEG sensors with respect to the head differ from subject to subject, and from session to session for the same subject. In this paper, a method is presented and tested to estimate the scalp MEG distributions from whole-head MEG measurements. The goal is to remove the discrepancy of MEG measurements caused by the various sensor positions with respect to the head, as well as to reduce the smearing effect caused by the distance of the MEG sensors from the scalp. The MEG measurement was first projected to a hypothetical dipole layer within the head volume conductor model using the inverse solution. The scalp MEG estimation was then obtained from the resultant dipole layer by the forward solution. The results from simulation studies, phantom experiments, and the auditory evoked field analysis demonstrated that, with reasonable signal to noise ratios, this method is a feasible way to achieve our goals. PMID- 10791685 TI - Differences in resting EEG related to ability. AB - The aim of the present study was to investigate the relationship between different EEG measures (mean power, mean frequency, approximated entropy and coherence), and ability (creativity and intelligence). For that purpose the EEG of 115 student-teachers (Intelligence: M= 115.17; SD = 12.78; IQ(min)= 82; IQ(max)= 136; Creativity - standardized scores: M = 55.97; SD = 10.67; C(min)= 38; C(max)= 84) was recorded while they were resting with eyes open and closed. The study showed only weak correlations between measures based on the level of activity in different areas (mean power, mean frequency and approximated entropy) and creativity. The correlations with IQ scores were even less pronounced. On the other hand, coherence measures showed a much more intense relationship both with creativity as well as with intelligence. In the eyes-open state these differences were mainly distributed over the right hemisphere. The results are discussed in the light of different theories relating brain functioning and ability. PMID- 10791686 TI - Growth factors in ulcer healing: lessons from recent studies. AB - Growth factors such as epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) and more recently vascular endothelial growth factor (VEGF) have been used extensively to heal experimental gastric, duodenal and colonic ulcers in animal models. Encouraging results have been reported in clinical trials with EGF and bFGF. Since our laboratory has been involved with the initial ulcer healing studies with bFGF, PDGF and VEGF, we summarize here the major lessons from these studies and from literature data. These conclusions relate to the role of: 1) gastrointestinal (GI) secretion; 2) epithelial versus vascular components of the healing; 3) efficacy in the upper and lower GI tract; 4) quality of ulcer healing; as well as 5) the endogenous origin; and 6) molar potency of growth factors. Namely, among these growth factors only EGF inhibits gastric acid and stimulates duodenal bicarbonate secretion, while chronic administration of bFGF slightly enhances gastric secretion and PDGF has no effect demonstrating that potent ulcer healing can be achieved without influencing acid base and mucus secretion. This might be related to the fact that these growth factors stimulate with varying potency virtually all the cellular elements needed for ulcer healing, e.g., epithelial cell proliferation and migration by EGF > bFGF > PDGF, fibroblast proliferation by bFGF > PDGF and angiogenesis by VEGF > bFGF >> PDGF >> EGF. Conceptually, the most interesting results were obtained recently with VEGF which is virtually specific for angiogenesis, illustrating that stimulation of vascular factors is sufficient for ulcer healing because epithelial cells apparently spontaneously proliferate and migrate over a dense granulation tissue to complete the healing process. Since these growth factors directly stimulate the cell components of ulcer healing, it is probably not surprising that they are active in both upper and lower GI tract lesions, produce good quality of ulcer healing in comparison with spontaneously healed duodenal ulcers which are hypovascular and muscle regeneration is not part of natural healing. Contrary to other antiulcer drugs, these growth factors are endogenously derived and play a role in the natural history of ulcer healing, and since these relatively large peptides (18-45 kDa) are active in ng quantities, their molar potency is 2-7 million times superior to cimetidine-like drugs. Thus growth factors are endogenously derived very potent antiulcer drugs which act independently of GI secretion, are active in upper and lower GI lesions, and since they stimulate virtually all the cells of the healing process, they produce an excellent quality of ulcer healing. PMID- 10791687 TI - Expression of cyclooxygenase (COX)-1 and COX-2 in adaptive cytoprotection induced by mild stress. AB - Prostaglandins (PG) derived from COX-1 play an important role in the maintenance of mucosal integrity but the role of COX-2-derived products in mucosal defence mechanism has not been fully explained. Mild stress is known to prevent gastric mucosal lesions induced by severe stress via the phenomenon of adaptive cytoprotection but it remains unknown which COX is involved in this adaptation. In this study, the mucosal expression of COX-1 and COX-2 was examined and the inhibitors of these enzymes were used to determine the contribution of these enzymes in adaptive cytoprotection induced by mild stress. Male Wistar rats were exposed to mild water immersion and restraint stress (WRS) at various time intervals ranging from 5 min up to 2 h followed 1 h later by exposure to severe 3.5 h WRS with or without pretreatment with: 1) NS-398 (10 mg x kg(-1) i.g.), a selective COX-2 inhibitor; 2) resveratrol (5 mg x kg(-1) i.g.), a selective COX-1 inhibitor; 3) meloxicam (2 mg x kg(-1) i.g.), preferential COX-2 inhibitor; and 4) indomethacin (5 mg x kg(-1) i.p), non-selective inhibitor of COX. The number of WRS lesions was counted, gastric blood flow (GBF) was measured by H2-gas clearance technique, mucosal biopsy samples were taken for the assessment of PGE2 by radioimmunoassay, and the expression of COX-1 and COX-2 mRNA by RT-PCR. WRS for 3.5 h produced numerous gastric lesions, decreased GBF by 48% and inhibited formation of PGE2 by 68% as compared to intact mucosa. Exposure to mild WRS during 5-30 min by itself failed to affect mucosal integrity but significantly attenuated gastric lesions induced by exposure to severe 3.5 h stress; the maximal protective effect being achieved with mild WRS during 15 min. This protective effect was accompanied by the rise in GBF and the generation of PGE2 in the gastric mucosa. After extension of mild WRS from 15 min up to 1 or 2 h before more severe 3.5 h WRS, the loss of cytoprotective effect of mild WRS against severe stress accompanied by significant fall in the GBF were observed. Pretreatment with NS-398 (10 mg x kg(-1) i.g.) that failed to affect mucosal PGE2 generation, reduced significantly the protection and accompanying rise in GBF produced by mild WRS whereas resveratrol partly reduced the protection and the rise in GBF induced by mild WRS. Meloxicam or indomethacin significantly inhibited PGE2 generation and completely abolished the hyperemia and protection induced by mild WRS against more severe stress. The protective and hyperemic effects of mild WRS were completely restored by the addition of 16,16 dm PGE2 (5 microg x kg(-1) i.g.) to NS-398 or resveratrol, while the deleterious effects of meloxicam and indomethacin were significantly attenuated by the concomitant treatment with this PGE2 analogue. We conclude that PG derived from both, COX-1 and COX-2 appear to be involved in adaptive cytoprotection developed in response to mild stressors. PMID- 10791688 TI - Antacid talcid activates in gastric mucosa genes encoding for EGF and its receptor. The molecular basis for its ulcer healing action. AB - In previous studies [Gut 35 (1994) 896-904], we demonstrated that antacid talcid (TAL) accelerates gastric ulcer healing and provides better quality of mucosal restoration within the scar than the omeprazole (OME). However, the mechanisms of TAL-induced ulcer healing are not clear. Since growth factors promote cell proliferation, re-epithelization, angiogenesis and ulcer healing, we studied whether TAL and/or OME affect expression of epidermal growth factor (EGF) and its receptors (EGF-R) in both normal and ulcerated gastric mucosae. Rats with or without acetic acid-induced gastric ulcers (n = 64) received i.g. twice daily 1 mL of either: A) placebo (PLA); B) TAL 100 mg; or C) OME 50 mg x kg(-1) for 14 d. Studies of gastric specimens: 1) ulcer size; 2) quantitative histology; 3) expression of EGF mRNAs was determined by RT/PCR; 4) gastric sections were immunostained with antibodies against EGF and its receptors. In non-ulcerated gastric mucosa of placebo or omeprazole treated group, EGF expression was minimal, while EGF-R was localized to few cells in the mucosal proliferative zone. Gastric ulceration triggered overexpression of EGF and its receptor in epithelial cells of the ulcer margin and scar. In ulcerated gastric mucosa TAL treatment significantly enhanced (versus PLA and omeprazole) expression of EGF and EGF-R. OME treatment reduced expression of EGF in ulcerated mucosa by 55 +/- 2% (P < 0.01). It is concluded that: 1) treatment with TAL activates genes for EGF and its receptor in normal and ulcerated gastric mucosae; 2) since EGF promotes growth of epithelial cells and their proliferation and migration, the above actions of TAL provide the mechanism for its ulcer healing action and improved (versus OME) quality of mucosal restoration. PMID- 10791689 TI - The antidepressant effect of an antiulcer pentadecapeptide BPC 157 in Porsolt's test and chronic unpredictable stress in rats. A comparison with antidepressants. AB - Various antidepressants have antiulcer activity. Likewise, the models currently used in ulcers and depression disorders research have a considerable degree of similarity. Therefore, the possibility that depression disorders could be effectively influenced by a primary antiulcer agent with a cyto/organoprotective activity, such as the novel stomach pentadecapeptide BPC 157, was investigated in two rat depression assays. First, a forced swimming test (a Porsolt's procedure) was used. As a more severe procedure, chronic unpredictable stress (after 5 d of unpredictable stress protocol, once daily drug application during stress procedure, open field-immobility test assessment at fourth or sixth day of medication) was used. In a forced swimming test, a reduction of the immobility time in BPC 157 (10 microg, 10 ng x kg(-1) i.p.) treated rats corresponds to the activity of the 15 mg or 40 mg (i.p.) of conventional antidepressants, imipramine or nialamide, respectively, given according to the original Porsolt's protocol. In chronic unpredictable stress procedure, particular aggravation of experimental conditions markedly affected the conventional antidepressant activity, whereas BPC 157 effectiveness was continuously present. The effect of daily imipramine (30 mg) medication could be seen only after a more prolonged period, but not after a shorter period (i.e., 4-d protocol). In these conditions, no delay in the effectiveness was noted in BPC 157 medication and a reduction of the immobility of chronically stressed rats was noted after both 4 and 6 d of BPC 157 (10 microg, 10 ng) medication. PMID- 10791690 TI - Gastric mucosal lesions induced by complete dopamine system failure in rats. The effects of dopamine agents, ranitidine, atropine, omeprazole and pentadecapeptide BPC 157. AB - Up to now, for gastric lesions potentiation or induction, as well as determination of endogenous dopamine significance, dopamine antagonist or dopamine vesicle depletor were given separately. Therefore, without combination studies, the evidence for dopamine significance remains split on either blockade of dopamine post-synaptic receptor or inhibition of dopamine storage, essentially contrasting with endogenous circumstances, where both functions could be simultaneously disturbed. For this purpose, a co-administration of reserpine and haloperidol, a dopamine granule depletor combined with a dopamine antagonist with pronounced ulcerogenic effect, was tested, and the rats were sacrificed 24 h after injurious agent(s) administration. Haloperidol (5 mg x kg(-1) b.w. i.p.), given alone, produced the lesions in all rats. Reserpine (5 mg x kg(-1) b.w. i.p.), given separately, also produced lesions. When these agents were given together, the lesions were apparently larger than in the groups injured with separate administration of either haloperidol or reserpine alone. Along with our previous results, when beneficial agents were co-administered, all dopaminomimetics (bromocriptine 10 mg, apomophine 1 mg, amphetamine 20 mg x kg( 1) i.p.) apparently attenuated the otherwise consistent haloperidol-gastric lesions. Likewise, an apparent inhibition of the reserpine-lesions was noted as well. However, if they were given in rats injured with combination of haloperidol and reserpine, their otherwise prominent beneficial effects were absent. Ranitidine (10 mg), omeprazole (10 mg), atropine (10 mg), pentadecapeptide BPC 157 (Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val) (10 microg or 10 ng x kg(-1) i.p.) evidently prevented both haloperidol-gastric lesions and reserpine-gastric lesions. Confronted with potentiated lesions following a combination of haloperidol and reserpine, these agents maintained their beneficial effects, noted in the rats treated with either haloperidol or reserpine alone. The failure of dopaminomimetics could be most likely due to more extensive inhibition of endogenous dopamine system activity, and need for remained endogenous dopamine for their salutary effect, whereas the beneficial activities of ranitidine, omeprazole, atropine, pentadecapeptide BPC 157 following dopamine system inhibition by haloperidol+reserpine suggest their corresponding systems parallel those of dopamine system, and they may function despite extensive inhibition of endogenous dopamine system activity. PMID- 10791691 TI - Stimulation of gastric acid secretion by progesterone metabolites as neuroactive steroids in anesthetized rats. AB - The effect of neuroactive progesterone metabolites, 5alpha- and 5beta-pregnan 3alpha-ol-20-one, and their stereoisomers at the 3 C site, 5alpha- and 5beta pregnan-3beta-ol-20-one, on gastric acid secretion was investigated in urethane anesthetized rats. Both 5alpha- and 5beta-pregnan-3alpha-ol-20-one dose dependently (0.3-3 mg x kg(-1), i.v.) stimulated gastric acid secretion with an early onset of action. Their potency and efficacy were almost the equivalent of one another. In contrast, their stereoisomers did not have a significant effect even at 10 mg x kg(-1) (i.v.). The 5beta-pregnan-3alpha-ol-20-one (3 mg x kg(-1), i.v.)-stimulated gastric acid secretion was remarkably inhibited by bilateral vagotomy or pretreatment with atropine (1 mg x kg(-1), i.v.). An antagonist of the GABA(A) receptor, picrotoxin, at 3 and 6 mg x kg(-1) (i.v.), significantly inhibited the 5beta-pregnan-3alpha-ol-20-one (3 mg x kg(-1), i.v.)-stimulated gastric acid secretion. These results indicate that naturally occurring neuroactive steroids, 5alpha- and 5beta-pregnan-3alpha-ol-20-one, stimulate gastric acid secretion in a stereoselective and dose-dependent manner in urethane anesthetized rats. It is likely that the action of these neuroactive steroids is of central origin and that interaction with GABA(A) receptors and stimulation of vagal pathway are involved in its mechanism of action. PMID- 10791692 TI - Alpha-2 adrenergic and opioid receptor-mediated gastroprotection. AB - Clonidine inhibited the development of gastric mucosal lesions induced by either acidified ethanol or indomethacin. The ED50 values were: 7.1 and 5.2 microg x kg( 1) orally, respectively. The gastroprotective effect was antagonised by the pre synaptic alpha-2 antagonist yohimbine, the more selective alpha-2 antagonist Ch 38083 and the pre-synaptic alpha-2B antagonist prazosin. Moreover, the non selective opioid receptor antagonist naloxone, the delta receptor selective naltrindole also reversed the clonidine-induced mucosal protective action. Clonidine was also effective following intracerebroventricular administration with the ED50 of 37 ng/rat against ethanol-induced mucosal damage. Our results suggest that: 1) the gastroprotective effect of clonidine is likely to be mediated by alpha-2B adrenoceptor subtype; 2) there is an interaction between pre synaptic alpha-2 adrenoceptors and opioid system; and 3) clonidine can induce gastroprotection by central mechanism. PMID- 10791693 TI - Vesicular monoamine transporters in the rat stomach. AB - Cellular distribution of vesicular monoamine transporters (VMATs), known to regulate vesicular storage and release of biogenic amines (i.e., catecholamines, serotonin, histamine, etc.), have been studied in the rat stomach using in situ hybridization histochemistry (ISHH) and immunohistochemical (IHC) techniques. 35S UTP labeled riboprobes showed that mRNAs of both VMATs are expressed in the gastric mucosa. A combination of ISHH and IHC verified that most of the parietal cells (among other epithelial cells) express mRNA of the peripheral type transporter (VMAT1) while enterochromaffin-like cells (ECL) of the fundic mucosa express mRNA of the central type (VMAT2). In addition, with double fluorescent IHC we detected VMAT1 protein in serotoninergic enterochromaffin cells (EC) of the stomach and in gastrin producing G cells of the antral mucosa. Similarly to the fundus, VMAT2 protein was present in ECL cells and in the enteric plexus. Surprisingly, serotonin- and/or histamine-containing cells in the connective tissue compartments of the stomach (i.e., lamina propria and submucosa), immunoreactive for a mast cell specific antigen, displayed neither VMATI nor VMAT2 immunoreactivity. Distribution of VMATs in the rat stomach support our previous observations on aminergic properties of two important gastrointestinal (GI) epithelial cell populations primarily known for other specific secretory products, i.e. dopaminergic properties of acid producing parietal cells and histaminergic properties of gastrin producing G cells. These data emphasize the existence of a non-neuronal, intrinsic aminergic system in the GI tract. PMID- 10791694 TI - Vagus-mediated activation of mucosal mast cells in the stomach: effect of ketotifen on gastric mucosal lesion formation and acid secretion induced by a high dose of intracisternal TRH analogue. AB - TRH analogue, RX 77368, injected intracisternally (i.c.) at high dose (3 microg/rat) produces gastric mucosal lesion formation through vagal-dependent pathway. The gastric mucosal hyperemia induced by i.c. RX 77368 was shown to be mediated by muscarinic vagal efferent fibres and mast cells. Furthermore, electrical vagal stimulation was observed to induce gastric mucosal mast cell degranulation. The aim of the study was to assess the influence of ketotifen, a mast cell stabilizer, on RX 77368-induced gastric lesion formation and gastric acid secretion. RX 77368 (3 microg, i.c.) or vehicle (10 microL, i.c.) was delivered 240 min prior to the sacrifice of the animals. Ketotifen or vehicle (0.9% NaCl, 0.5 mL) was injected intraperitoneally (i.p.) at a dose of 10 mg x kg(-1) 30 min before RX 77368 injection. The extent of mucosal damage was planimetrically measured by a video image analyzer (ASK Ltd., Budapest) device. In the gastric acid secretion studies, the rats were pretreated with ketotifen (10 mg x kg(-1), i.p.) or vehicle (0.9% NaCl, 0.5 mL, i.p.), 30 min later pylorus ligation was performed and RX 77368 (3 microg, i.c.) or vehicle (0.9% NaCl, 10 microL, i.c.) was injected. The rats were killed 240 min after i.c. injection, and the gastric acid secretion was measured through the titration of gastric contents with 0.1 N NaOH to pH 7.0. RX 77368 (3 microg, i.c.) resulted in a gastric mucosal lesion formation involving 8.2% of the corpus mucosa (n = 7). Ketotifen elicited an 85% inhibition on the development of mucosal lesions (n = 7, P < 0.001) whereas ketotifen alone had no effect on the lesion formation in the mucosa (n = 7). The RX 77368 induced increase of gastric acid secretion was not influenced by ketotifen pretreatment in 4-h pylorus-ligated animals. Central vagal activation induced mucosal lesion formation is mediated by the activation of mucosal mast cells in the stomach. Mast cell inhibition by ketotifen does not influence gastric acid secretion induced by i.c. TRH analogue in 4-h pylorus ligated rats. PMID- 10791695 TI - Modulation by GABA(B) and delta opioid receptors of neurally induced responses in isolated guinea-pig taenia coli and human colonic circular muscle. AB - The GABA-ergic and opioid modulation of neurally induced muscle responses was studied in isolated guinea-pig taenia coli and human colonic circular muscle, using identical field stimulation parameters (rectangular pulses of 0.5 ms duration, 9 V x cm(-1) intensity, trains of 3 pulses at 0.5 Hz, repeated every 1/3/5 min). The stimulation-induced contractions were inhibited in both preparations by GABA and baclofen; the IC50 values in human colonic circular muscle were approximately 100 and 31.0 microM, respectively. In guinea-pig taenia coli, the inhibition by 10(-4) M GABA was dose-dependently reversed by 10(-4)-10( 3) M of GABA(B) receptor antagonist CGP 35348; antagonism by phaclofen was less effective in the same concentration range. In human colonic circular muscle, inhibition by 3 x 10(-5) M baclofen was fully reversed by 10(-3) M CGP 35348. With the exception of caecum, the delta 2 opioid receptor agonist deltorphin II was a potent inhibitor in human colonic circular muscle. 10(-8) M Deltorphin caused a 74.4 +/- 9.6% (n = 4) inhibition which was reversed by 10(-6) M of delta receptor selective peptide antagonist BOC-Tyr-Pro-Gly-Phe-Leu-Thr(OtBu). Deltorphin II was ineffective in guinea-pig taenia coli even at 10(-6) M; the same concentration caused an 84.3 +/- 7.9 (n = 4) inhibition in human preparations. It is concluded that: 1) GABA-ergic modulatory mechanisms are present both in human colonic circular muscle and guinea-pig taenia coli; 2) the GABA receptors involved are of type B; and 3) delta opioid receptor-mediated modulation functions only in human colonic circular muscle in regions other than the caecum. PMID- 10791696 TI - Helicobacter pylori. One bacterium and a broad spectrum of human disease! An overview. AB - Since the historical rediscovery of gastric spiral Helicobacter pylori in the gastric mucosa of patients with chronic gastritis by Warren and Marshall in 1983, peptic ulcer disease has been largely viewed as being of infectious aetiology. Indeed, there is a strong association between the presence of H. pylori and chronic active gastritis in histology. The bacterium can be isolated in not less than 70% of gastric and in over 90% of duodenal ulcer patients. Eradication of the organism has been associated with histologic improvement of gastritis, lower relapse rate and less risk of bleeding from duodenal ulcer. The bacterium possesses several virulence factors enabling it to survive the strong acid milieu inside the stomach and possibly damaging host tissues. The sequence of events by which the bacterium might cause gastric or duodenal ulcer is still not fully elucidated and Koch's postulates have never been fulfilled. In the majority of individuals, H. pylori infection is largely or entirely asymptomatic and there is no convincing data to suggest an increase in the prevalence of peptic ulcer disease among these subjects. An increasingly growing body of literature suggests an association between colonization by H. pylori in the stomach and a risk for developing gastric mucosa-associated lymphoid tissue (MALT), MALT lymphoma, gastric adenocarcinoma and even pancreatic adenocarcinoma. The bacterium has been implicated also in a number of extra-gastrointestinal disorders such as ischaemic heart disease, ischaemic cerebrovascular disease, atherosclerosis, and skin diseases such as rosacea, but a causal role for the bacterium is missing. Eradication of H. pylori thus seems to be a beneficial impact on human health. Various drug regimens are in use to eradicate H. pylori involving the administration of three or four drugs including bismuth compounds, metronidazole, clarithromycin, tetracyclines, amoxycillin, ranitidine, omeprazole for 1-2 weeks. The financial burden, side effects and emergence of drug resistant strains due to an increase in the use in antibiotics for H. pylori eradication therapy need further reconsideration. PMID- 10791697 TI - 13C-Urea breath test is superior in sensitivity to detect Helicobacter pylori infection than either antral histology or rapid urease test. AB - There is no single technique which fulfils the criterion for a reference method to detect Helicobacter pylori (Hp) infection. The aim was to compare the results of antral histology (H), rapid urease test (U) and urea breath test (UBT) from antral biopsy samples in patients having gastric or duodenal lesions during upper GI endoscopy. We used the following methods: 1) biopsy specimens for histology (Warthin-Starry staining); 2) rapid urease test; and 3) 13C-urea breath test with infrared spectrometry. The total number of patients was 166 examined by H, U, and UBT. H, U and UBT were negative (-) in 64 patients and positive (+) in 51. The true positivity and false negativity (%, number of patients in parentheses) of each method based upon the positivity of the other two tests were: H+, U+ (54): UBT+, 94.4% (51) and UBT-, 5.6% (3); H+, UBT+ (57): U+, 89.5% (51) and U-, 10.5% (6); U+, UBT+ (65): H+, 78.5% (51) and H-, 21.5% (14). If Hp infection is considered to be positive when at least two tests detect the presence of Hp, UBT shows the highest sensitivity in comparison to histology of biopsy specimens and urease test. UBT is highly recommended as a screening test for Hp infection in patients presenting upper GI endoscopic alterations. PMID- 10791699 TI - Decrease of serum carotenoids in Crohn's disease. AB - Crohn's disease (CD) is frequently complicated by various nutritional disturbances. Although it is important to correct these disturbances, the nutritional status of CD patients has been poorly documented, especially concerning vitamin status. The aims of this study were (a) to measure the serum concentrations of vitamin A and six other carotenoids (lutein, zeaxanthin, alpha , beta-carotene, alpha-, beta-cryptoxanthin) in patients with CD and to compare them with those in healthy controls and (b) to follow the changes of serum carotenoid levels in CD patients during treatment. Twenty-eight patients with CD and 23 healthy persons were included in this study. The results of twelve patients were followed up through one year. The patients were free of any nutritional treatment. The serum concentrations of carotenoids were measured with high-pressure liquid chromatography (HPLC). The serum concentrations of five carotenoids were significantly lower in the patients than in the controls (vitamin A, zeaxanthin: P < 0.001; alpha-, beta-carotene: P < 0.01; lutein: P < 0.05). The carotenoid status of the followed patients advanced to the normal range, but this increase was not significant. These findings suggest that there is a deficiency of vitamin A and its provitamins in Crohn' s disease prior to treatment. However, because we did not evaluate the vitamin intake in this study, we could not conclude which of the factors--poor intake, increased requirement, or malabsorption--was more important in decreasing of carotenoid levels. PMID- 10791698 TI - Probiotic bacteria and intestinal health: new methods of investigation. AB - This paper highlights some new methods in the probiotic research based on the use of colonic biopsies and molecular biological techniques for strain identification. PMID- 10791700 TI - The tumoricidal effect of Trypanosoma cruzi: its intracellular cycle and the immune response of the host. AB - Many experimental evidences indicate that infection with Trypanosoma cruzi delays or inhibits the growth of malignant tumors in different strains of mice and in rats. These facts were verified by different workers. Although earlier workers proposed that this effect would be due to a toxin of T. cruzi, most of the accumulated evidences do not agree with such proposal. This present hypothesis agrees with the experimental data and proposes that the liberation of many endocellular antigens by destruction of some cancer cells, infected with T. cruzi, gives rise to an autoimmune response against antigens of analogous cancer cells, which limits or inhibits tumor growth. This point of view is supported by experimental studies on Chagas' disease which showed the role of T. cruzi, to induce autoimmune reactions against target organs of the disease. On the basis of this hypothesis I postulate a new way to stimulate the immune system of the host against cancer. PMID- 10791701 TI - Transactional conflict between psychobiology and culture in the etiology of postpartum depression. AB - In vulnerable individuals, postpartum depression may be triggered by the disruption of neuropsychological processes during pregnancy and parturition. It is proposed that in Western societies pre- and postnatal psychological and instrumental needs are poorly met and this crisis may interact with neurophysiological factors in compromising maternal psychological functioning. The influence of developmental factors and psychological and physical stressors are discussed in relation to women's neuropsychological health during the peripartal transition. It is hypothesized that the environmental milieu may support, neglect or derail the mother's psychobiological transition from pregnancy to effective postnatal parenting and resumption of family and work roles. PMID- 10791702 TI - What are the implications of the interaction between DDT and estrogen receptors in the body? AB - The organochlorine pesticide, dichlorodiphenyltrichloroethane (DDT), which is fat soluble and persistent in the body and environment, has estrogenic activity. There has been an apparent association with breast cancer, which has implicated DDT binding with estrogen receptors (ERs). The mechanism of DDT-ER interaction at target sites is similar to estrogen, with protein synthesis resulting in an estrogenic response. Other than the female reproductive sites, DDT could possibly bind to ERs present in other body systems. The recent discovery of a beta receptor has introduced a new understanding of estrogen and DDT binding. An understanding of the molecular biology of the DDT-ER interaction in breast tissue could possibly explain the risk of breast cancer. Estrogen and other estrogenic compounds compete with DDT by their estrogenic potential. DDT-ER interaction in the body has wider implications in terms of its genotoxic potential and role in carcinogenesis. PMID- 10791704 TI - Herpesviruses in human periodontal disease. AB - Recent studies have identified various herpesviruses in human periodontal disease. Epstein-Barr virus type 1 (EBV-1) infects periodontal B-lymphocytes and human cytomegalovirus (HCMV) infects periodontal monocytes/ macrophages and T lymphocytes. EBV-1, HCMV and other herpesviruses are present more frequently in periodontitis lesions and acute necrotizing ulcerative gingivitis-lesions than in gingivitis or periodontally healthy sites. Reactivation of HCMV in periodontitis lesions tends to be associated with progressing periodontal disease. Herpesvirus associated periodontitis lesions harbor elevated levels of periodontopathic bacteria, including Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteriodes forsythus, Prevotella intermedia, Prevotella nigrescens and Treponema denticola. It may be that active periodontal herpesvirus infection impairs periodontal defenses, thereby permitting subgingival overgrowth of periodontopathic bacteria. Alteration between latent and active herpesvirus infection in the periodontium might lead to transient local immunosuppression and explain in part the episodic progressive nature of human periodontitis. Tissue tropism of herpesvirus infections might help explain the localized pattern of tissue destruction in periodontitis. Absence of herpesvirus infection or viral reactivation might explain why some individuals carry periodontopathic bacteria while still maintaining periodontal health. Further studies are warranted to delineate whether the proposed herpesvirus-periodontopathic bacteria model might account for some of the pathogenic features of human periodontal disease. PMID- 10791703 TI - Antigen of erythroblast (Ag-Eb): a membrane protein that may be an erythroid specific transferrin receptor. AB - Only a limited number of erythroid cell surface markers have been described in the literature. Ag-Eb was originally described as an erythroid-specific cell surface glycoprotein and could be used as an erythroid differentiation marker, but more recent studies suggest this localization is more widespread. From the data summarized in this review, it is hypothesized that Ag-Eb is a member of a subset of the transferrin receptor family and that it functions together with these receptors in the uptake and metabolism of iron, particularly at histo hematic barriers. PMID- 10791705 TI - Herpesvirus in localized juvenile periodontitis. AB - Herpesvirus genomic sequences can be detected in gingival crevicular fluid of adult periodontitis lesions. Herpesviruses are immunosuppressive and may facilitate establishment of subgingival pathogens. Electron microscopic studies have identified nuclear and cytoplasmic virus-like inclusions in gingival inflammatory cells from localized juvenile periodontitis (LJP). The present study aimed to determine if herpesviruses occur in LJP lesions and if human cytomegalovirus (HCMV) activation is associated with elevated levels of subgingival Actinobacillus actinomycetemcomitans, the putative bacterial pathogen of LJP. Eleven systemically healthy patients exhibiting LJP (10-23 yr) were studied. In each patient, subgingival samples were pooled from 3 periodontitis lesions around first molar and incisor teeth (5-11 mm periodontal pocket depth) and from 3 gingivitis/healthy sites around canines (2-3 mm periodontal pocket depth). Polymerase chain reaction (PCR) was used to detect herpesvirus DNA and HCMV cDNA of major capsid protein transcripts, indicative of viral activation. Selective culture and 16S rRNA PCR were used to identify A. actinomycetemcomitans. Of 11 deep periodontal samples, 8 showed HCMV, 7 showed Epstein-Barr virus type 1 (EBV-1), 1 showed EBV type 2, 6 showed herpes simplex virus (HSV) and 8 showed viral co-infection. Of 11 shallow periodontal samples, 2 showed HCMV, 2 showed EBV-1, 1 showed HSV and 2 showed viral co-infection. The difference in occurrence of HCMV and viral co-infection between deep and shallow periodontal sites was statistically significant (p =0.031). HCMV activation was detected in deep pockets of all 5 virally positive patients with early LJP (aged 10-14 years) but only in 1 of 3 virally positive LJP patients older than 14 years, and not in any shallow pocket tested. HCMV activation appeared related to absence of radiographic crestal alveolar lamina dura, a possible indication of periodontal disease progression. A. actinomycetemcomitans tended to be more prevalent in samples showing active than latent HCMV infection. The present findings are consistent with the notion that periodontal herpesvirus infection and possibly HCMV activation constitute important features of the etiopathogenesis of LJP. PMID- 10791706 TI - Morphology of azurophil lysosomes in polymorphonuclear leukocytes from humans with rapidly progressive periodontitis. AB - Previous investigations have shown that subjects with rapidly progressive periodontitis (RPP), an early-onset aggressive form of periodontitis, have polymorphonuclear leukocytes (PMNs) with increased intracellular levels of beta glucuronidase, a characteristic enzyme of azurophil lysosomes. The current study attempted to account for that increase. Ten healthy controls and 10 otherwise healthy subjects with RPP participated. PMNs from peripheral blood were separated, fixed and reacted for peroxidase to identify azurophil lysosomes. Using transmission electron microscopy, 20 PMNs per subject were photographed at 10,000x. Photographs were subsequently digitized and analyzed by computer. RPP PMNs had a higher percentage of the area of the cell profile occupied by azurophil lysosomes compared to control subjects' PMNs. The RPP subjects also had greater absolute numbers of azurophil lysosomes per cell. Lysosome shape was assessed visually. There were no differences between RPP and control groups for lysosome shape, with the majority of lysosomes in each group exhibiting a round or oval shape. RPP lysosomes did exhibit a significantly greater mean size. PMID- 10791707 TI - Profile of subgingival microbiota in children with primary dentition. AB - Eruption of primary teeth has a great influence on the oral environment by providing suitable niches for bacterial colonization. The aim of the study was to investigate the composition of the subgingival microbiota of primary incisors, canines and molars in 40 systemically healthy children aged 4-5 yr, chosen randomly. Subgingival plaque samples were taken from the mesiobuccal sites of primary incisors (61, 81), canines (53, 73) and molars (64, 84). The samples were cultured for bacterial isolation anaerobically and in 10% CO2 plus air using selective and non-selective media. Forty-one different microbial species were isolated. Gemella morbillorum and Peptostreptococcus magnus were statistically significantly more frequently detected in incisors while P. micros, Streptococcus intermedius, Bacteroides forsythus, Fusobacterium nucleatum, Prevotella loeschei, P. melaninogenica and Selenomonas sputigena were more frequently detected in molars. The bacterial species S. constellatus, G. morbillorum and P. magnus were isolated in greater numbers in incisors and P. micros, S. intermedius, Campylobacter concisus, Bacteroides egertheii, B. forsythus, P. oralis and S. sputigena were isolated in greater numbers in molars, respectively. Cluster analysis revealed 4 clusters in which 6-7 bacterial species were elevated above mean levels. Cluster I was predominated by S. constellatus, S. mitis, S. sanguis, G. morbillorum, P. melaninogenica and P. oralis; cluster II was predominated by S. sanguis, Actinomyces naeslundii, Capnocytophaga gingivalis, C. ochracea and P. intermedia; cluster III was predominated by S. mitis, C. ochracea, F. nucleatum, P. loeschei, P. melaninogenica and P. oralis; and finally cluster IV was predominated by S. sanguis, C. gingivalis, Veillonella parvula, Campylobacter gracilis, F. nucleatum and P. intermedia. The bacterial species S. constellatus, P. micros, Pseudoramibacter alactolyticus, Eikenella corrodens and F. nucleatum were associated with non-bleeding sites while S. intermedius, C. concisus, P. intermedia and P. loescheii were found more frequently in bleeding sites. PMID- 10791708 TI - Storage conditions of avulsed teeth affect the phenotype of cultured human periodontal ligament cells. AB - After severe injury to the periodontal ligament (PL), the phenotypes of cells recolonizing root surfaces influence the extent and type of repair processes. In teeth that are replanted following avulsion injury, recolonization of the PL space by osteogenic cells instead of by PL fibroblasts may favor bone formation (i.e. ankylosis) instead of PL regeneration. We consider here that recolonization processes depend in part on the storage conditions of the teeth following avulsion. We used an in vitro cell culture model to assess the effect of storage conditions on immunohistochemical staining of several marker proteins that are expressed by osteogenic cells (osteopontin and alkaline phosphatase) and fibroblasts (alpha-smooth muscle actin, type III and XII collagens). Prior to cell culture, extracted human premolar teeth were stored in air ("dry") or in alpha-MEM ("wet") for either 30 or 120 min as surrogate conditions for the variations of extra-alveolar tooth storage that may occur following avulsion. Collagenase/trypsin-digested suspensions of PL cells were prepared from the tissue adherent to the extracted root surface. Passage #2 or #3 cultures were immunostained and examined by fluorescence microscopy. For type XII collagen, cells from wet samples displayed perinuclear staining while cells from 30-min dry samples showed only isolated foci. The staining for 120-min dry samples was weak and non-specific. alpha-Smooth muscle actin was not incorporated into stress fibers in wet samples, whereas dry samples demonstrated prominent stress fibers stained for alpha-smooth muscle actin. Detached cytoplasmic fragments resembling cell processes that stained for alpha-smooth muscle actin were abundant in dry samples, indicating the presence of highly contractile cells. The staining for osteopontin was mainly perinuclear but was more intense in dry samples. The focal adhesion pattern of osteopontin staining in 120-min dry samples resembled that of migrating osteogenic cells. The pattern of staining did not vary for type III collagen or alkaline phosphatase, although staining for alkaline phosphatase was more intense in samples stored under dry conditions. We conclude that prolonged extra-alveolar dry storage favors increased in vitro growth of contractile cells expressing osteogenic cell markers while storage in cell culture medium favors growth of cells with the classical phenotype of PL fibroblasts. PMID- 10791709 TI - Cyclosporin A inhibits production and activity of matrix metalloproteinases by gingival fibroblasts. AB - Cyclosporin A (CyA) is a potent immunosuppressor used in organ transplantation and in the management of various autoimmune diseases. Gingival overgrowth is one of the side-effects of the CyA-treatment, affecting the attached gingiva of 25 81% of treated patients. To investigate the production and activity of matrix metalloproteinases (MMPs) in the CyA-induced gingival overgrowth, 2 well documented models were utilized: the in vivo CyA-induced rat gingival overgrowth and primary cultures of human gingival fibroblasts treated with CyA. Our results obtained from the Western blot assays demonstrated clearly that the production of MMP-1 and MMP-3 was significantly inhibited by CyA at similar concentrations found in the serum of patients undergoing CyA-treatment. Moreover, the gelatinolytic activity of MMP-2 was also reduced both in cultured fibroblasts and in the rat CyA-induced gingival overgrowth. Taken together, the data presented here suggest that these inhibitory effects may contribute to the extracellular matrix (ECM) components accumulation in the CyA-induced gingival overgrowth. PMID- 10791710 TI - The heat shock response of Escherichia coli. AB - A large variety of stress conditions including physicochemical factors induce the synthesis of more than 20 heat shock proteins (HSPs). In E. coli, the heat shock response to temperature upshift from 30 to 42 degrees C consists of the rapid induction of these HSPs, followed by an adaptation period where the rate of HSP synthesis decreases to reach a new steady-state level. Major HSPs are molecular chaperones, including DnaK, DnaJ and GrpE, and GroEL and GroES, and proteases. They constitute the two major chaperone systems of E. coli (15-20% of total protein at 46 degrees C). They are important for cell survival, since they play a role in preventing aggregation and refolding proteins. The E. coli heat shock response is positively controlled at the transcriptional level by the product of the rpoH gene, the heat shock promoter-specific sigma32 subunit of RNA polymerase. Because of its rapid turn-over, the cellular concentration of sigma32 is very low under steady-state conditions (10-30 copies/cell at 30 degrees C) and is limiting for heat shock gene transcription. The heat shock response is induced as a consequence of a rapid increase in sigma32 levels and stimulation of sigma32 activity. The shut off of the response occurs as a consequence of declining sigma32 levels and inhibition of sigma32 activity. Stress-dependent changes in heat shock gene expression are mediated by the antagonistic action of sigma32 and negative modulators which act upon sigma32. These modulators are the DnaK chaperone system which inactivate sigma32 by direct association and mediate its degradation by proteases. Degradation of sigma32 is mediated mainly by FtsH (HflB), an ATP-dependent metallo-protease associated with the inner membrane. There is increasing evidence that the sequestration of the DnaK chaperone system through binding to misfolded proteins is a direct determinant of the modulation of the heat shock genes expression. A central open question is the identity of the binding sites within sigma32 for DnaK, DnaJ, FtsH and the RNA polymerase, and the functional interplay between these sites. We have studied the role of two distinct regions of sigma32 in its activity and stability control: region C and the C-terminal part. Both regions are involved in RNA polymerase binding. PMID- 10791711 TI - Survival of bacteria during oxygen limitation. AB - Regulatory mechanisms that enable bacteria associated with food, drinks and the human body to adapt to changes in the availability of oxygen are reviewed. Excess oxygen induces two adaptive responses to oxidative stress. Five or more control circuits enable enteric bacteria to generate energy and grow well in anaerobic environments. Two sets of enzymes catalyse both nitrate and nitrite reduction, and dual two-component regulatory systems sense and respond to the available nitrate and nitrite in the environment. The periplasmic nitrate reductase enables bacteria to scavenge low concentrations of nitrate: similar systems are found in food-borne and other pathogens. PMID- 10791712 TI - Starvation and osmotic stress induced multiresistances. Influence of extracellular compounds. AB - Growth restriction due to stasis and/or hyperosmolarity is a common situation encountered by microorganisms in nature. Therefore, they have developed defence systems allowing them to withstand these periods. Bacteria respond to these conditions by a metabolic reprogramming which leads to a cellular state of enhanced resistance. This communication reviews recent advances in knowledge of the molecular basis of this phenomenon in different bacteria. PMID- 10791713 TI - Regulation of stress response in Oenococcus oeni as a function of environmental changes and growth phase. AB - Oenococcus oeni is a lactic acid bacterium which is able to grow in wine and perform malolactic fermentation. To survive and grow in such a harsh environment as wine, O. oeni uses several mechanisms of resistance including stress protein synthesis. The molecular characterisation of three stress genes hsp18, clpX, trxA encoding for a small heat shock protein, an ATPase regulation component of ClpP protease and a thioredoxin, respectively, allow us to suggest the existence in O. oeni of multiple regulation mechanisms as is the case in Bacillus subtilis. One common feature of these genes is that they are expressed under the control of housekeeping promoters. The expression of these genes as a function of growth is significantly different. Surprisingly, the clpX gene, which is induced by heat shock, was highly expressed in the early phase of growth. In addition to stress protein synthesis, adaptation to the acid pH of wine requires efficient cellular systems to extrude protons. Using inhibitors specific for different types of ATPases, we demonstrated the existence of H+-ATPase and P-type ATPase. PMID- 10791714 TI - Trehalose reserve in Saccharomyces cerevisiae: phenomenon of transport, accumulation and role in cell viability. AB - Strains of Saccharomyces cerevisiae deleted for TPS1 encoding trehalose-6 phosphate synthase still accumulate trehalose when harbouring a functional MAL locus. We demonstrate that this accumulation results from an active uptake of trehalose present in the 'yeast extract' used to make the enriched culture media and that no accumulation is observed in mineral media. The uptake of trehalose was shown to be mediated by the alpha-glucoside transporter encoded by AGT1, the expression of which is linked to the presence of a functional MAL locus. Deletion of this gene in a MAL+ tps1 mutant abolished trehalose accumulation on a maltose or galactose mineral medium. However, small amounts of disaccharide were still detected in a agt1 tps1 double mutant when the medium was supplemented with 10 g trehalose l(-1), indicating the existence of a non-concentrative low-affinity sugar transporter. The presence of the high-affinity trehalose permease allowed us to investigate the effect of increasing exogenous trehalose from 0 to 10 g(-1) on intracellular accumulation. A maximum of ca. 10% (wt/wt dry cells) trehalose was attained in the presence of only 1 g l(-1) of disaccharide in the medium. The capability to monitor the intracellular content of trehalose by varying its extracellular concentration, independent of genetic alterations of the trehalose metabolic machinery, allowed the remarkable contribution of this molecule in stress tolerance to be demonstrated, as the higher the trehalose content, the longer the cell survival to a severe heat shock and to glucose starvation. PMID- 10791715 TI - Basic features of the stress response in three species of bifidobacteria: B. longum, B. adolescentis, and B. breve. AB - The presence of the dnaK heat shock gene could be demonstrated for B. longum NCC481, B. longum NCC490, B. longum NCC585, B. adolescentis NCC251, and B. breve NCC298. Induction of dnaK on the transcriptional level was shown for NCC251 and NCC481 by increasing temperatures. NCC251 showed an additional chaperone induction after salt or bile-salt treatment. In both strains preconditioning with bile-salts protected against otherwise lethal concentrations thereof. NCC251 when subjected to a heat stress was able to survive an otherwise lethal temperature (55 degrees C). Cross-protection was demonstrated for NCC251 since salt pretreatment resulted in increased tolerance after freeze-thawing cycles or lethal heat stress. PMID- 10791716 TI - Fatty acid membrane composition and activation of glycine-betaine transport in Lactococcus lactis subjected to osmotic stress. AB - Lactococcus lactis subsp. cremoris NCDO763 accumulates glycine-betaine (betaine) when submitted to an osmotic stress with NaCl. Betaine transport activity increases with the extent of the osmotic upshock but also with growth temperature, and supplementation of the medium by Tween-80. Fatty acid analysis of the lipid fraction of L. lactis NCDO763 reveals significant modifications of the fatty acid composition of the membrane when cells are submitted to osmotic stress, high temperature or Tween-80 medium supplementation. The main modification in L. lactis membrane fatty acid composition in response to high osmolality is the increase of Cyclopropane Fatty Acid (CFA) deltaC19:0, whereas Unsaturated/Saturated ratio remains unchanged. PMID- 10791717 TI - Impact of thermal variations on biochemical and physiological traits in Pectinatus sp. AB - The influence of temperature on cellular fatty acid composition and on heat stress tolerance was studied in the two species of Pectinatus, an anaerobic gram negative bacterium. Cellular fatty acid (FA) patterns were determined for Pectinatus species cultivated in MRS medium at various defined conditions of temperature and pH. Our study shows that fluctuations of growth temperature and pH induced important changes in the ratio of unsaturated FAs (UFAs) to saturated FAs (SFAs). The major differences in the FA composition as a function of growth temperature concerned C15:0 and C17:0 for the SFAs and C15:1 and C17:1 for the UFAs. The most significant adaptation of lipid composition to lower growth temperatures was the strong increase of UFAs, particularly for C15:1 and C17:1 concomitantly with a decrease of SFAs (C15:0 and C17:0). When the pH of the culture medium was lowered from 6.2 to 4.0, a notable drop in the synthesis of the UFAs C15:1 and C17:1 was observed together with an important increase of C18 cyclopropane (C18-cyc) and high carbon number SFAs. Thermal modifications also provoked changes in Pectinatus behaviour. We observed that P. cerevisiiphilus was more heat sensitive than P. frisingensis. Mild exponential phase cells were treated for 1 h, at 40 degrees C for P. cerevisiiphilus or at 41 degrees C for P. frisingensis. This thermal adaptation induced tolerance against heat challenge (49 and 50 degrees C for P. cerevisiiphilus and P. frisingensis, respectively). Survival of P. cerevisiiphilus and P. frisingensis adapted cells was, respectively, 3400- and 790-fold higher than control. Interestingly, adapted cells of P. cerevisiiphilus were more thermotolerant than P. frisingensis pretreated cells. PMID- 10791718 TI - Demonstration of a protein synthesis in starved Campylobacter jejuni cells. AB - For many years, environmental microbiologists working on water samples, have reported differences between bacterial counts performed by culture and by microscopy. These observations have led to the demonstration of the viable but non-culturable (VNC) state in bacteria. Some hygienist specialists underlined the risk presented by pathogenic bacteria in the VNC state. The VNC state in bacteria has been studied by a number of authors, but the relation between VNC state and bacterial stress response has not been established yet, while the VNC state is generally described in responses to adverse conditions. Campylobacter jejuni enter the VNC state in response to starvation. In our study, we searched for a protein synthesis in the first hours of the cell starvation exposure. Three Campylobacter jejuni strains were suspended in filtered, sterilized, distilled water, and incubated at 4 degrees C with gentle shaking (100 rpm). After 1, 2, 3, 4 and 5 h of starvation, C. jejuni cells were removed and subjected to a heat shock (55 degrees C, 3 min) and to a conductimetric assay. Results obtained showed that a protein synthesis occurred in the onset of the starvation period, and that these improved the nutrient assimilation and enhanced the heat resistance in starved cells. PMID- 10791719 TI - Increase of the ATP-dependent phosphoenolpyruvate carboxykinase activity in Sinorhizobium meliloti (Rhizobium meliloti) during hypothermic environmental conditions. AB - Sinorhizobium meliloti growth is affected when the incubation temperature is lower than 22 degrees C. In culture media containing glucose or fructose (1%, w/v), the doubling time at 19 degrees C was about 6.25 h during the exponential growth phase, while it was 2.75 h at 30 degrees C; at 17 degrees C it was three fold higher than at 30 degrees C. Modifications in the bacterial metabolism explain the doubling time increase when bacteria are incubated at low temperature. We determine here, the phosphoenolpyruvate carboxykinase (PEPCK) activity increases when S. meliloti cells first grown at 30 degrees C are shifted at 17 degrees C and incubated for 10 h at this low temperature; we noted the PEPCK activity was three-fold higher in cells incubated in media containing glucose and shifted from 30 to 17 degrees C than in cells maintained at 30 degrees C, while it was only 1.5-fold higher in cells grown in media containing fructose. PMID- 10791720 TI - Effect of temperature on growth characteristics of Bacillus cereus TZ415. AB - The effect of temperature on the maximal specific growth rate was studied in Bacillus cereus between 5 and 40 degrees C cultivated in courgette broth and rich medium (J broth). B. cereus grown from 5 to 38 degrees C in rich medium. No growth was observed in courgette broth below 10 degrees C. The Arrhenius plot was fitted from experimental data of B. cereus grown in rich medium and at regulated pH, oxygen and temperature. Two domains which are separated by a critical temperature around 13 degrees C can be distinguished with regard to temperature dependence of maximal specific growth rate. Over the cold domain from 5 to 13 degrees C, the temperature characteristic was 2.6 fold higher than over the sub optimal domain from 13 to 38 degrees C suggesting that the growth temperature regulates several metabolic pathways. PMID- 10791721 TI - Effect of temperature and physiological state on the fatty acid composition of Pseudomonas aeruginosa. AB - The influence of temperature and physiological state on fatty acid profiles of cell membranes of a gram-negative bacteria was studied in this work. It has been shown that fatty acid composition is largely modified by these two parameters. Lipids play an important role in the composition and the function of cell membranes. These modifications of membrane structures are very important to understand because of their consequences on cell viability. PMID- 10791722 TI - Lactococcus lactis, a bacterial model for stress responses and survival. AB - The dairy organism, Lactococcus lactis, is continuously exposed to stress conditions generated during industrial processes. To identify the mechanisms that confer resistance to the lethal effects of oxygen and thermal stress, we isolated resistant strains by insertional mutagenesis. Mutated genes were identified and mutations were shown to confer resistance to multiple stresses (including non selected stresses such as carbon starvation). Our results revealed that metabolic flux plays an important role in L. lactis stress response, and suggested that phosphate and guanine pools may be intracellular stress sensors. As previously shown, we also observed an increase of stress resistance during the stationary phase. We have evidence that stationary phase actually initiates very early during growth. Taken together, these data show that the stationary phase is a very complex system with multiple participants interacting altogether. These results reinforce the idea of the interdependence of stress response and the intimate relation between metabolic flux and stress responses in L. lactis. PMID- 10791723 TI - Identification of general stress genes in Enterococcus faecalis. AB - The susceptibility and the acquisition of tolerance of E. faecalis ATCC 19433 to heat, ethanol, bile salts, NaCl, H2O2 and pH shifts were determined. During exposure to these environmental stresses, protein synthesis analysed by 2-D electrophoresis revealed 167 stress proteins. Six stress protein were found to be induced by at least six of the eight treatments and considered to be general stress proteins (Gsp). Western blotting identified two of these Gsp as DnaK and GroEL. Analysis of the four other Gsp revealed that at least two of them were not previously described. PMID- 10791724 TI - Quantifying the hurdle concept by modelling the bacterial growth/no growth interface. AB - The hurdle concept described eloquently over many years by Professor Leistner and his colleagues draws attention to the interaction of factors that affect microbial behaviour in foods. Under some circumstances these effects are additive. Under others the implication is that synergistic interactions lead to a combined effect of greater magnitude than the sum of constraints applied individually. Predictive modelling studies on the combined effects of temperature and water activity and temperature and pH suggest that the effect of these combinations on growth rate is independent. Where the effect of the two factors is interactive rather than independent is at the point where growth ceases--the growth/no growth interface. An interesting and consistent observation is that a very sharp cut off occurs between conditions permitting growth and those preventing growth, allowing those combinations of factors to be defined precisely and modelled. Growth/no growth interface models quantify the effects of various hurdles on the probability of growth and define combinations at which the growth rate is zero or the lag time infinite. Increasing the stringency of one or more hurdles at the interface by only a small amount will significantly decrease the probability of an organism growing. Understanding physiological processes occurring near the growth/no growth interface and changes induced by moving from one side of the interface to the other may well provide insights that can be exploited in a new generation of food preservation techniques with minimal impact on product quality. PMID- 10791725 TI - Characterization and analysis of a new gene involved in glucose starvation response in Enterococcus faecalis. AB - The genome sequence of Enterococcus faecalis, led us to discover that gls24, encoding a general stress protein, seems to be in the second last position of a putative six-gene operon structure. Interestingly, another gene named orf4 located just upstream from gls24 shows strong identity (72%) with this last. To determine the role of the orf4 gene in E. faecalis, we have constructed a mutant strain by homologous recombination. Phenotypic analysis of these cells, reveals that Orf4 is probably involved in cell structure in stationary phase. PMID- 10791726 TI - A specific mutation in Saccharomyces cerevisiae adenylate cyclase, Cyr1K176M, eliminates glucose- and acidification-induced cAMP signalling and delays glucose induced loss of stress resistance. AB - The cAMP-protein kinase A (PKA) pathway in the yeast Saccharomyces cerevisiae plays a major role in the control of metabolism, proliferation and stress resistance. Derepressed cells show a rapid increase in the cAMP level (within 1 min) after addition of glucose or after intracellular acidification. A specific mutation in adenylate cyclase, the enzyme that catalyzes the synthesis in cAMP, largely prevents both cAMP responses. The responsible mutation was originally called lcr1 (for lack of cAMP responses); lcr1 was later identified as allelic with CYR1/CDC35. The mutation was introduced into the CYR1 gene of a W303-1A wild type strain, which resulted in a large decrease in cAMP signalling. Furthermore, there was a strong reduction in GTP/Mg2+-stimulated but not in Mn2+-stimulated adenylate cyclase activity in isolated plasma membranes, which is consistent with the absence of signalling through adenylate cyclase in vivo. Glucose-induced activation of trehalase was reduced and mobilization of trehalose and glycogen and loss of stress resistance were delayed in the lcr1 mutant. Because of the absence of cAMP signalling during exponential growth on glucose, it was concluded that glucose-induced cAMP signalling is restricted to the transition from gluconeogenic/respiratory to fermentative growth. Activation of the PKA pathway is mediated by a G protein (either Ras1/Ras2 or Gpa2). Constitutive activation of the pathway by Ras2val19 or Gpa2val132 has a negative effect on glycogen and trehalose accumulation and heat shock survival. The lcr1 mutation partially suppresses this effect indicating that the target sites of the two G-proteins on adenylate cyclase might have at least a part in common. PMID- 10791727 TI - Cloning, sequencing and characterization of the ccpA gene from Enterococcus faecalis. AB - Enzymes involved in the metabolism of complex carbon and energy sources are unnecessary under conditions of abundant, readily metabolisable nutrients such as glucose or fructose. The repression of these enzymes by glucose has been termed carbon catabolite repression. Mechanisms involved in the carbon catabolite repression in gram-positive bacteria are known to differ from those of gram negative bacteria such as Escherichia coli. It appears to be mediated by transcriptional repression, requiring trans-acting CcpA, a member of the LacI GalR family of bacterial regulatory proteins and a cis-acting consensus sequence, designated cre. Here, we report the cloning and characterisation of the chromosomal ccpA gene from Enterococcus faecalis JH2-2. This gene is predicted to encode a 333 amino acids protein with nearly 75% identity to CcpA of Lactobacillus casei. PMID- 10791728 TI - Effects of high hydrostatic pressure on membrane proteins of Salmonella typhimurium. AB - Salmonella typhimurium is a leading cause of foodborne diseases. Today high hydrostatic pressure treatments are considered as alternative methods of preservation. To select optimal conditions of treatment, we have to characterize the cell targets of pressure. In this study the action of pressure on the bacterial membrane proteins is analysed. The total membrane extract is obtained by lysis of cells separated by equilibrium density gradient centrifugation. Protein content is analysed by electrophoresis SDS-PAGE and visualised by silver stain. Electrophoretic profiles reveal the presence of three major outer membrane proteins and 12 minor proteins in control bacteria outer membranes. Outer membrane protein content is drastically modified after treatments. In some cases, except for the major proteins OmpA and LamB, other outer membrane proteins seem to totally disappear. LamB is more resistant to hyperbaric exposure when the pH of the media is acidic. This behaviour could be explained by a different conformation adopted by the LamB protein depending on the extracellular pH. This work allows us to define membrane proteins as a target of high hydrostatic pressure treatments. Knowledge of the behaviour of these bacterial membrane proteins subjected to pressure under different conditions (pH, temperature, a(w)...) could allow an increase in the efficiency of treatments. PMID- 10791729 TI - Acid responses of Listeria monocytogenes. AB - The acid response and the correlated protein synthesis in Listeria monocytogenes were studied. The lowest pH value which L. monocytogenes could resist was dependent on the strain and the kind of acid used. Previous adaptation to an intermediary pH augmented bacterial resistance to a subsequent lethal acidic pH. The acid tolerance was also growth phase dependent. Organic volatile acids exerted a more deleterious effect on L. monocytogenes than inorganic acids, because weak acids infer a lower intracytoplasmic pH. The responses to acid adaptation (mildly acidic pH) and acid stress (lethal acidic pH) shared the majority of acid-induced proteins. The bacteria required more stress proteins to face severe acidic conditions. In order to obtain insights into the role these acid-induced proteins play in the mechanism of acid resistance, the proteins were analysed by two-dimensional electrophoresis and the most abundant were identified by peptide mass fingerprinting and MALDI-TOF mass spectrometry. PMID- 10791730 TI - Growth of Lactococcus lactis strains at low water activity: correlation with the ability to accumulate glycine betaine. AB - Lactococcus lactis strains were divided into two groups based on their ability to grow in the presence of an upper limit of either 2% w/v NaCl (sensitive) or 4% w/v NaCl (tolerant). Growth inhibition of NaCl tolerant strains was substantially relieved by glycine betaine which was accumulated in significant amounts when growing at low water activities (a(w)). Very little accumulation of glycine betaine occurred during growth of the NaCl sensitive strains. The NaCl tolerant strains had substantial levels of glycine betaine transport activity in vitro, whereas the NaCl sensitive strains had little or no such activity. A low a(w) sensitive mutant of L. lactis subsp. cremoris MG1363 (NaCl tolerant) was isolated following ISS1 insertional mutagenesis. This mutant was inhibited at an a(w) of 0.988 produced by addition of 2% w/v NaCl or the equivalent glucose concentration (0.58 M). The mutant did not accumulate glycine betaine when growing at low a(w), and did not transport glycine betaine when assayed in vitro. PMID- 10791731 TI - Mechanism of action of pulsed high electric field (PHEF) on the membranes of food poisoning bacteria is an 'all-or-nothing' effect. AB - Salmonella typhimurium (CRA 1005) was more sensitive than Listeria monocytogenes (NCTC 11994) to pulsed high electric field (PHEF) treatment in distilled water (10, 15 and 20 kV/cm), 10 mM tris-maleate buffer pH 7.4 (15 kV/cm) and model beef broth (0.75% w/v: 15 kV/cm). Sublethal injury could not be detected using a selective medium plating technique, indicating that bacterial inactivation by PHEF may be an 'all-or-nothing' event. PHEF-induced membrane permeabilization resulted in increased UV-leakage and a decreased ability of L. monocytogenes to maintain a pH gradient. PMID- 10791732 TI - Biochemical and biophysical studies of Bacillus subtilis envelopes under hyperosmotic stress. AB - The behaviour and state of the envelopes from B. subtilis cultures grown in Luria Bertani (LB) medium with and without 1.5 M NaCl are compared. Under hypertonic conditions, the hydrophobicity of the cultures increases. The phospholipid and fatty acid (FA) compositions show important differences: a higher cardiolipin (CL) content [at the expense of phosphatidylglycerol (PG)], and a higher unsaturated and straight chain FA content. The fluidity of the membranes, determined with fluorescent probes, indicates an increase in viscosity of the cytoplasmic membrane. The consequences of these variations in membrane permeability and osmotolerance are discussed. PMID- 10791733 TI - Influence of different factors on the inactivation of Salmonella senftenberg by pulsed electric fields. AB - The influence of growth phase, cell concentration, pH and conductivity of treatment medium on the inactivation of Salmonella senftenberg by high electric field pulses (HELP) was studied. Cells were more resistant to HELP treatments at the beginning of the logarithmic phase and at the stationary phase. Microbial inactivation was not a function of the initial cell concentration. At constant input voltage, electric field strength obtained in the treatment chamber depended on medium conductivity. At the same electric field strength, conductivity did not influence S. senftenberg inactivation. At the same conductivity, inactivation of S. senftenberg was bigger at neutral than acidic pH. PMID- 10791734 TI - The effect of osmo-induced stress on product formation by Zymomonas mobilis on sucrose. AB - The intensification of biosynthesis of fructooligosaccharides in the presence of high salt concentrations was observed during sucrose (10%) fermentation by Zymomonas mobilis 113S. A 0.6 M NaCl concentration led to an increase of oligosaccharide productivity by 3.5-fold. Sorbitol formation was increased in the presence of 0.16 M NaCl and was inhibited at highest salt concentrations. In a medium with high (65%, w/w) sucrose content the salts gave inhibitory effects on fructooligosaccharide production by lyophilised Z. mobilis cells. Influence of salts on gluconic acid and sorbitol formation under these conditions was studied. The ratio of oligosaccharides and gluconic acid productivity (Qolig./Qglucon.) was increased approximately 2 times at 1% KCl. Sorbitol formation was not significantly influenced in the presence of KCl (up to 2%). PMID- 10791735 TI - Proteins variations in Listeria monocytogenes exposed to high salinities. AB - Listeria monocytogenes Scott A grown in the minimal chemically defined medium M6LT was challenged to a concentration of either 35 or 65 g l(-1) of NaCl for 1 h in the presence of a [35S]cysteine-[35S]methionine labelling mix. The protein patterns were analysed by 2D-electrophoresis in the two conditions and isoosmotic condition (5 g l(-1) of NaCl in M6LT). A great number of proteins which were synthesized under isoosmotic conditions were either completely repressed or expressed at a reduced level, at 65 g l(-1) and to a lesser extent at 35 g l(-1) of NaCl. At 35 g l(-1) of NaCl, six proteins were up-regulated, five proteins showed no change in expression level and five were repressed. Among the proteins up-regulated at 35 g l(-1) of NaCl, a single one (18.7 kDa, pI 5.05) was up regulated at 65 g l(-1) too. We observed 21 proteins which were repressed at 65 g l(-1) of NaCl, among which 11 completely disappeared. Some of the up-regulated proteins have characteristics of molecular weight and isoelectric point close to those of stress proteins reported elsewhere: the protein induced both at 35 and 65 g l(-1) might correspond to a previously proposed universal stress protein of Listeria. Some proteins which were repressed at 65 g l(-1) have molecular weights close to those of virulence proteins. PMID- 10791736 TI - Quaternary ammonium compound stresses induce specific variations in fatty acid composition of Pseudomonas aeruginosa. AB - The involvement of cell membrane fatty acids in resistance of Pseudomonas aeruginosa to Quaternary Ammonium Compounds (QACs) stresses was investigated. The strain was grown in a medium with increasing concentrations of different biocides: two QACs, and two non-QACs. In the presence of two QACs only, the strain was able to grow with increasing concentrations. During cellular adaptation to QACs, the resistance to the same biocide increased. A principal component analysis was performed with whole of fatty acid compositions which highlighted a specific variation for the cultures in presence of QACs. These modifications gave evidence of the outer membrane involvement in cellular response to the presence of QACs. PMID- 10791737 TI - Metabolism of Lactococcus lactis subsp. cremoris MG 1363 in acid stress conditions. AB - The metabolism of glucose by Lactococcus lactis subsp. cremoris MG 1363 remains homolactic whatever the pH of the culture medium. The growth rate decreased with the acidification of the medium until a limit pH value of 4.0 for which no growth was observed. In contrast, the specific rate of glucose consumption decreased only for very low pH values, i.e., below 4.5. The efficiency of biomass synthesis relative to the energy supply decreased when the medium pH diminished, as illustrated by Y(ATP) values. This observation was related to the increase in both components of the proton-motive force when the pH decreased. The growth stopped when the internal pH reached a limit value of 5.4 due to organic acid accumulation. PMID- 10791738 TI - Glycerol metabolism in Lactobacillus collinoides: production of 3 hydroxypropionaldehyde, a precursor of acrolein. AB - Lactobacillus collinoides is a lactic acid bacterium commonly found in fermenting apple juice. Although this bacterium is not particularly involved in malolactic conversion, the presence of L. collinoides in cider may have serious consequences on the product. L. collinoides is indeed considered to be responsible for the transformation of glycerol to 3-hydroxypropionaldehyde (3-HPA), a precursor of acrolein that spoils the product quality by generating bitter tastes. The purpose of our work was to evaluate the influence of environmental and culture conditions on the conversion of glycerol to 3-HPA in L. collinoides, and to obtain a DNA probe of the gene coding for glycerol dehydratase, the enzyme responsible for this conversion. PMID- 10791739 TI - Is the accumulation of osmoprotectant the unique mechanism involved in bacterial osmoprotection? AB - Sucrose, trehalose, maltose, cellobiose, gentiobiose, turanose and palatinose are very unusual osmoprotectants for Sinorhizobium meliloti, because these compounds, unlike other bacterial osmoprotectants, do not accumulate as cytosolic osmolytes in salt-stressed S. meliloti cells. Rather, these compounds were catabolized during early exponential growth, and contributed to enhance the cytosolic levels of the two endogenously-synthesized osmolytes: glutamate and the dipeptide N acetylglutaminylglutamine amide (NAGGN). Furthermore, all of the disaccharides that acted as powerful osmoprotectants shared the same uptake routes in S. meliloti. Here, we show that these disaccharides, in fact, belong to a new family of non-accumulated sinorhizobial osmoprotectants and that two mechanisms of osmoprotection coexist in S. meliloti. PMID- 10791740 TI - Taking injuries of surviving bacteria into account for optimising heat treatments. AB - The main assets of early conventional models applied in the field of canned food industries are their simplicity and their robustness. Moreover, a certain standardisation of these models allows the intrinsic quantification of a food process like sterilisation, regardless of the nature of concerned microbial populations. However, a first drawback of conventional models is their monofactorial nature: only temperature is considered for the evaluation of microbial heat resistance. A second limit of early survival models is that conventional estimates of heat resistance are made by recovering heated surviving cells at optimal incubation conditions. However, many investigators observed that culture medium and incubation temperature influence both the ratio of injured cell recovery and estimated heat resistance values. Mafart and Leguerinel [Mafart, P., Leguerinel, I., 1998. Modeling combined effects of temperature and pH on heat resistance of spores by a linear-Bigelow equation. J. Food Sci. 63, 6 8] developed a model describing the heat resistance of spores as a function of temperature and pH which is an extension of the Bigelow equation. A short while later, they added a further term to their model in order to consider the water activity of the heating medium (Gaillard, S., Leguerinel, I., Mafart, P., 1998. Model for combined effects of temperature, pH and water activity on thermal inactivation of Bacillus cereus spores. J. Food Sci. 63, 887-889). From their model, the authors proposed an extension of the concept of biological destruction value (BDV) noted L(T, pH, a(w)), that they called primary BDV. More recently, we developed a second multifactorial model describing the effect of the temperature, the pH and the water activity of the recovery medium on the estimated D-value of heated spores (unpublished). From this new model we propose the concept of secondary BDV noted L'(T, pH, a(w)). We show that, for calculations of heat processes, the effective BDV, M, to be considered is the overall function M = LL'. From another model [Mafart, P., Leguerinel, I., 1997. Modelling the heat stress and the recovery of bacterial spores. Int. J. Food Microbiol. 37, 131 135], it can be shown that the secondary BDV is also the corrective factor to be considered to assess the overall decimal reduction ratio q: q = L'n where n is the conventional decimal reduction ratio without taking into account the effect of environmental factors of the recovery medium on the effective heat resistance of injured spores. PMID- 10791741 TI - Basic aspects of food preservation by hurdle technology. AB - Hurdle technology is used in industrialized as well as in developing countries for the gentle but effective preservation of foods. Previously hurdle technology, i.e., a combination of preservation methods, was used empirically without much knowledge of the governing principles. Since about 20 years the intelligent application of hurdle technology became more prevalent, because the principles of major preservative factors for foods (e.g., temperature, pH, a(w), Eh, competitive flora), and their interactions, became better known. Recently, the influence of food preservation methods on the physiology and behaviour of microorganisms in foods, i.e. their homeostasis, metabolic exhaustion, stress reactions, are taken into account, and the novel concept of multitarget food preservation emerged. In the present contribution a brief introduction is given on the potential hurdles for foods, the hurdle effect, and the hurdle technology. However, emphasis is placed on the homeostasis, metabolic exhaustion, and stress reactions of microorganisms related to hurdle technology, and the prospects of the future goal of a multitarget preservation of foods. PMID- 10791742 TI - Characterization of a new set of mutants deficient in fermentation-induced loss of stress resistance for use in frozen dough applications. AB - In frozen dough applications a prefermentation period during the preparation of the dough is unavoidable and might also be important to obtain bread with a good texture. A major disadvantage of the prefermentation period is that it is associated with a rapid loss of the freeze resistance of the yeast cells. A major goal for the development of new baker's yeast strains for use in frozen dough applications is the availability of strains that maintain a better freeze resistance during the prefermentation period. We have isolated mutants that retain a better stress resistance during the initiation of fermentation. Some of these showed the same growth rate and fermentation capacity as the wild type cells. These mutants are called 'fil', for deficient infermentation induced loss of stress resistance. First we used laboratory strains and heat stress treatment, given shortly after the initiation of fermentation, as the selection protocol. The first two mutants isolated in this way were affected in the glucose activation mechanism of the Ras-cAMP pathway. The fil1 mutant had a partially inactivating point mutation in CYR1, the gene encoding adenylate cyclase, while fil2 contained a nonsense mutation in GPR1. GPR1 encodes a member of the G protein coupled receptor family which acts as a putative glucose receptor for activation of the Ras-cAMP pathway. In a next step we isolated fil mutants directly in industrial strains using repetitive freeze treatment of doughs as selection protocol. Surviving yeast strains were tested individually for maintenance of fermentation capacity after freeze treatment in laboratory conditions and also for the best performing strains in frozen doughs prepared with yeast cultivated on a pilot scale. The most promising mutant, AT25, displayed under all conditions a better maintenance of gassing power during freeze-storage. It was not affected in other commercially important properties and will now be characterised extensively at the biochemical and molecular level. PMID- 10791744 TI - Modelling the growth of Listeria monocytogenes in dynamic conditions. AB - A recurrent neural network for the prediction of Listeria monocytogenes growth under pH and a(w) variable conditions was developed. The use of this model offered the possibility to take into account the consequences of the variations of the factors on L. monocytogenes growth. The effects of solutions, such as NaCl, acetic acid and NaOH, and their interactions on the response of L. monocytogenes cells were studied. Furthermore, the results showed the capacity of the recurrent neural network to predict growths carried out in different experimental conditions without using those used for its elaboration. PMID- 10791743 TI - Assessment of viability of microorganisms employing fluorescence techniques. AB - Viability assessment of microorganisms is relevant for a wide variety of applications in industry, including evaluation of inactivation treatments and quality assessment of starter cultures for beer, wine, and yoghurt production. Usually, the ability of microbial cells to reproduce is considered as the benchmark method for determination of cell viability, and this is most commonly determined by the plate count method. The time needed to form visible colonies, however, is relatively long. Therefore, there is an increasing interest in rapid methods which exploit criteria other than reproduction. In this review the applications of fluorescent probes for, e.g., determination of membrane integrity, enzyme activities, respiration, membrane potential and intracellular pH, are discussed in detail. PMID- 10791745 TI - Characterization of Streptococcus thermophilus strains that undergo lysis under unfavourable environmental conditions. AB - The autolysis of starter lactic acid bacteria appears as a promising way to enhance the flavour of fermented dairy products. The present work was aimed at investigating the autolysis phenomenon in Streptococcus thermophilus, a thermophilic lactic acid bacteria involved in the starters used for the production of yoghurts, Italian and Swiss-type cheeses. Out of 146 strains screened for their aptitude to spontaneously lyse at the end of growth in M17 medium containing lactose in limited concentration, six strains, among which is the type strain CNRZ 1358, were found to be highly autolytic. These autolytic strains are characterized by a typical bell-shaped growth curve. Lysis of the type strain, which was studied as the model, was triggered under unfavourable environmental conditions, such as lactose depletion and NaCl or organic solvents addition. The lysogenic character of this strain was evidenced. Taken together, our results indicate that the autolytic phenotype in S. thermophilus is linked to the lysogenic character but does not result from the massive prophage induction under stressing conditions. PMID- 10791746 TI - Changes in acid tolerance of Lactococcus lactis during growth at constant pH. AB - Cells of Lactococcus lactis MG1363 growing in batch culture in TYG (tryptone, yeast extract, glucose) medium at constant pH 7.0 became gradually more acid sensitive shortly after inoculation until a point of maximum sensitivity was reached in early log-phase. The acid tolerance then gradually increased in the mid- and late-log phase until maximum tolerance was reached at the onset of stationary phase. This pattern has been termed the growth-phase acid tolerance. The variation in acid tolerance seen in pH 7.0 grown cells of L. lactis MG1363 did not result from changes in internal pH or membrane H+ ATPase activity levels. Neither the amount of glucose present during mid-log phase nor the amount of lactate produced by the cells correlated with the pattern of the log-phase acid tolerance. Cells grown in partially spent TYG medium showed a reduced growth rate and increased acid tolerance compared to cells grown in fresh TYG medium. Supplementing the spent medium with tryptone or yeast extract or both restored the growth rate and cells became more sensitive to acid. Fractionation of tryptone yielded a fraction which stimulated the growth of MG1363 in partially spent medium and delayed the acquisition of acid tolerance. The active compound(s) has a putative molecular weight of about 1 kDa and was partially degraded by papain and trypsin. PMID- 10791747 TI - Relationship between the apparent heat resistance of Bacillus cereus spores and the pH and NaCl concentration of the recovery medium. AB - Conventional heat resistance data, D values, were previously established by other workers at optimal condition for spores outgrowth. However, in canned food conditions of outgrowth are generally suboptimal in term of pH, salt concentration, water activity. The combined effects of pH and NaCl level of the recovery medium for the D value and z(pH) value were studied. Spores of Bacillus cereus were heated at 95 degrees C in phosphate-citrate buffer media at pH 7. Cells were recovered at 25 degrees C in Nutrient Agar with pH ranging from 5 to 7 and 1% to 4% (w/w) NaCl concentration. For each condition D' values (decimal reduction time associated with the recovery media characteristics) were determined. The results show a major influence of the recovery pH on the D' values. This effect is characterised by the z'(pH) values, distance of recovery medium pH from optimum recovery pH* medium (6.7) which leads to a tenfold reduction time of D value. The increase of the salt concentration leads to a slight decrease of D' value. However z'(pH) values are not significantly affected by the salt concentration. A simple three parameter model describing the effects of pH and NaCl concentration of the recovery medium upon the heat resistance of spores is proposed. The interaction between pH and salt concentration is sufficiently low to be neglected by the model. PMID- 10791748 TI - Survival of a Pseudomonas fluorescens and Enterococcus faecalis aerosol on inert surfaces. AB - The aim of the work presented here is to study airborne bacteria survival on three inert supports (glass, polyvinyl chloride, and stainless steel). Two Pasteur Institute bacteria were selected: Enterococcus faecalis 10.30.15 and Pseudomonas fluorescens 56.90. We have observed that bacterial aerosol lethality increased proportionally with the relative humidity of the environment, the gram negative rod appearing more fragile than the cocci. A significant difference in survival rate is measured dependent on the supports tested, the greatest lethality being observed on the PVCs. PMID- 10791749 TI - Influence of culture pH on survival of Lactobacillus reuteri subjected to freeze drying. AB - L. reuteri was cultivated at pH 5 and 6. The cells were harvested at 0.5, 2.5 and 4.5 h after entering the stationary phase and their viability after freeze-drying was compared with their viability prior to freeze-drying. The highest viability- approximately 80% of the viability prior to freeze-drying--was obtained for cells from the pH 5 cultures harvested 2.5 h after entering the stationary phase. The time after entering the stationary phase did not influence the viability of the cells from the pH 6 cultures where the viability was approximately 50% irrespective of harvest time. Product formation was the same for pH 5 and pH 6 grown cells, whereas the pH 6 grown cells exhibited a more elongated morphology. PMID- 10791750 TI - Evaluation of biohazards in dehydrated biofilms on foodstuff packaging. AB - Plastic materials used for food packaging are clean but not sterile when the food is just packaged. Accidental wet contamination may occur at every moment between packaging and opening by the consumer: on polyethylene (PET), bacteria may adhere strongly and constitute a biofilm in less than 24 h. By rolling on themselves, PET sheets may contaminate food. We tried to show that contact with salted foodstuffs favoured microbial recovery. Four strains were chosen to perform biofilms on PET: Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Escherichia coli. Biofilms were dried up 24 h. Biofilm bacteria were stressed by adhesion, by starvation and by dehydration. However, they were capable of recovery in salted solutions or media, probably because one (or more) stress protected them against another stress. Stress was demonstrated by stress protein production, by mean of electrophoresis, and membrane lesions by mean of flow cytometry. Stress recovery was performed in aqueous salted solutions or salted brain-heart infusion with NaCl 9, 15, 20 and 30 g/l. Staphylococci were more sensitive to these stresses and recovery was a function of salt concentration. Gram-negative bacteria were little affected by stresses; salt effects were less important. If all these biofilms were capable of recovery from stresses in salted media, flexible PET could possibly lead to a health hazard when it is used for wet salt meats, e.g. PMID- 10791751 TI - Cellular activities in ultra-violet killed Escherichia coli. AB - In this work we analyze the physiological state of cells after lethal-UV dose disinfection using independent metabolic markers. Through the detection of some metabolic activities we proved that cell lysis does not immediately follow death in UV-irradiated Escherichia coli K12 cells. PMID- 10791752 TI - Adaptation of Lactobacillus alimentarius to environmental stresses. AB - Lactobacillus alimentarius BJ33 has been tested for its biopreservative capacities to improve quality and safety in many meat products. The combination of different preservatives such as NaCl, glucono-delta-lactone and citric acid with this protective culture during the manufacture of sausages represent an interesting alternative to control microbial spoilage and to extend product shelf life. The use of these preservatives may also limit the growth of L. alimentarius. In this study, the sublethal doses of these preservatives were determined and tested in combination to verify if the organism was able to adapt to these stresses. The sublethal doses of gluconic acid, citric acid, and NaCl were 100-110 mM, 50-55 mM and 8%, respectively. When the culture was first grown in MRS broth containing citric acid (50 or 55 mM) or gluconic acid (100 or 110 mM) and then transferred in MRS broth containing NaCl (8%), only limited growth was observed (O.D.(600 nm) = 0.2-0.3) after 6 days at 30 degrees C. However, when the culture was first grown in NaCl and then transferred in MRS broth containing gluconic or citric acid, growth was observed after 1 day (O.D.(600 nm) = 0.4-0.5) and after 5 days an O.D.(600 nm) of 0.8 was reached. Cell filamentation was also observed under electron microscopy when cells were grown for 2 days in presence of gluconic and citric acid at their sublethal doses and with a combination of 18 mM gluconic acid and 37 mM citric acid, but cellular elongation was not observed with cultures exposed to 8% NaCl. These results suggest that two different adaptation mechanisms are induced in L. alimenatrius when treated with organic acids and NaCl. PMID- 10791754 TI - Identification of genes responsible for improved cryoresistance in fermenting yeast cells. AB - Using repetitive freezing and thawing, different mutant industrial Saccharomyces cerevisiae strains with increased freeze resistance have been isolated. To get a better insight in the mechanisms responsible for this elevated resistance and to give us the opportunity to modify other strains so that they become more suitable for use in frozen dough preparations, we applied the microarray technology in order to identify genes that are differentially expressed in a freeze-resistant mutant when compared to a freeze-sensitive industrial yeast strain. PMID- 10791753 TI - Synergistic lethal combination of nitrite and acid pH on a verotoxin-negative strain of Escherichia coli O157. AB - This study was concerned with the possible consequences of reducing the nitrite concentration of a fermented sausage environment on the survival of the pathogen E. coli O157:H45, a verotoxin-negative relative of E. coli O157:H7. A liquid medium, FM, was constructed with a liquid phase, a(w) and pH similar to fermented sausage. Survival of E. coli O157:H45 in FM depended on both pH and nitrite concentration. In trials in which the pH was decreased by growing Pediococcus acidilactici in FM, survival of E. coli O157:H45 was clearly dependent on nitrite concentration; at least 100 ppm nitrite was required to inhibit growth and the number of survivors after 2 days with 200 ppm nitrite was 1000-fold less than in the absence of nitrite. In laboratory-scale sausage fermented with P. acidilactici, E. coli O157:H45 failed to grow in the absence of nitrite and the numbers slowly declined over 14 days. However, the rate of decline was much faster with nitrite present even at 50 ppm; at 200 ppm nitrite, the E. coli O157:H45 population declined 100 times faster than in the absence of nitrite. PMID- 10791755 TI - Survival and recovery of viable but noncultivable forms of Campylobacter in aqueous microcosm. AB - Previous studies suggesting that the persistence of thermotolerant Campylobacter in water, especially as a viable but non-cultivable form (VNC), was involved in human campylobacteriosis, the capacities of survival and resuscitation of a significant collection of 85 strains in aqueous microcosms were investigated. Two thirds of these strains (68%) were not detectable on agar medium after a stay of 14-21 days, whereas 21% reached this state before 14 days and 11% were non cultivable after a stay of 21 days. Some strains remained cultivable after 35 days in a shaken aqueous microcosm and beyond 60 days without shaking. After 30 days, 51% of the non-detectable strains by conventional culture were recovered after injection in 9-day fertilised chicken eggs. A kinetic study showed that the age of the viable but non-cultivable forms and characteristics of the strains could explain the variations of recovery. These results suggest that viable but non-cultivable forms of Campylobacter could be a potential risk of colonisation of human or animals and that an embryonic factor seems to be essential to allow resuscitation. PMID- 10791756 TI - Membrane fluidity of stressed cells of Oenococcus oeni. AB - The determination of membrane fluidity in whole cells of Oenococcus oeni was achieved by membrane probe 1,6-diphenyl-1,3,5-hexatriene fluorescence anisotropy measurements. The results demonstrated instantaneous fluidity variations with cells directly stressed during the measure. Heat (42 degrees C) or acid (pH 3.2) shocks decreased the anisotropy values (fluidising effects), whereas an ethanol shock (10% ethanol, v/v) increased the membrane rigidity. The velocities of fluidity variation with non-adapted or adapted cells (incubation in inhibitory growth conditions) were compared. The adaptation of the cells to acid conditions had no effect on the membrane fluidity variation after an acid shock. In contrast, the rates of membrane fluidity variation of adapted cells were 5- and 3 fold lower after a heat shock and an ethanol shock, respectively. These results suggest the positive effect of an adaptation on the membrane response and can help to explain the mechanisms of stress tolerance in Oenococcus oeni. PMID- 10791757 TI - Influence of thermal and osmotic stresses on the viability of the yeast Saccharomyces cerevisiae. AB - This work studies the effect of thermal and dehydration kinetics on the viability of Saccharomyces cerevisiae. The influence of the rate of temperature (T) and osmotic pressure (pi) increases are first investigated. Results showed that yeast viability is preserved by slow variations of temperature or osmotic pressure in a precise range of T or pi. The influence of a previous thermal stress on the resistance to a hyperosmotic stress is also studied. Temperatures equal to or lower than 10 degrees C allowed the preservation of viability after an osmotic stress whereas temperatures above 10 degrees C did not preserve yeast survival. PMID- 10791758 TI - Problems associated with the direct viable count procedure applied to gram positive bacteria. AB - Despite the numerous advantages of fluorescent in situ hybridization (FISH) for identifying a single bacterial cell with 16S rRNA probes, problems are encountered with starving bacteria in natural samples. The original direct viable count procedure (DVC) includes a revivification step in the presence of an antibiotic inhibiting cell division. Cells elongate and accumulate ribosomes. This results in a natural amplification of 16S rRNA molecules (target of FISH). However, it is limited to gram-negative bacteria which are sensitive to nalidixic acid. The objective of this study was to develop a procedure for estimating the number of metabolically active gram-positive Staphylococcus aureus and Enterococcus faecalis cells by the use of a method which combines the number of substrate-responsive cells and their identification by FISH. It was observed that no single published DVC method could apply to taxonomically different gram positive bacteria. Since cells were not counted, the revivification step in presence of nalidixic acid will be referred to as revivification without cell division. For each species, different low-nutrient media and complex media, different fluoroquinolones and beta-lactam antibiotics, concentrations of antibiotics, combinations of antibiotics, temperature and time were evaluated using bacteria in different physiological states and in natural samples. Enumeration of bacteria by plate counts and direct FISH were compared. The improved procedure should yield information about the physiological state, the taxonomic identity, and the enumeration of viable gram-positive bacteria. The application of DVC to an entire ecosystem is presently still a challenge. PMID- 10791759 TI - Modifications to methods for the enumeration and detection of injured Escherichia coli O157:H7 in foods. AB - Reliable methods are required for the detection and enumeration of potentially injured E. coli O157 in food in the presence of outnumbering competing bacteria. Selective agents can prevent or inhibit the recovery and subsequent multiplication of injured cells and direct inoculation, either into selective enrichment broths or onto selective agar plates is still used in many methods for E. coli O157 detection and enumeration. When compared with tryptone soya agar (TSA), sorbitol MacConkey agar (SMAC) was shown to underestimate the concentration of viable E. coli O157:H7 subjected to low pH and high NaCl concentration. Using a resuscitation stage on TSA followed by membrane transfer to SMAC improved recovery to levels obtained on TSA. The membrane method was used to monitor the numbers of artificially contaminated E. coli O157:H7 during the fermentation of a meat product and demonstrated better survival when compared to counts on SMAC. Six rapid methods for the detection of E. coli O157 in food (BAX E. coli O157, Reveal 8 E. coli O157-H7 screening test, VIP EHEC, VIDAS E. coli O157 (ECO), EHEC-Tek and Tecra E. coli O157 visual immunoassay), were evaluated using beetburgers, parsley and fermented meat artificially contaminated with injured cells. Methods using direct selective enrichment, with or without an elevated incubation temperature gave false-negative results. The incorporation of a non-selective pre-enrichment medium improved the detection rates of these assays by up to ten fold. PMID- 10791760 TI - Fluorescence assessment of Lactococcus lactis viability. AB - The reproduction and activity of lactic acid bacteria (LAB) are essential in their applications in the dairy industry and other fermentations. Traditionally used methods like plate counting and acidification tests require long incubation times and provide limited information. Fluorescence techniques provide possibilities for rapid assessment of cell physiology. We used traditional and fluorescence assays to assess the physiological condition of L. lactis subsp. lactis ML3 cultures that were exposed to various stress conditions. After exposure to some of the stress conditions, carboxyfluorescein (cF) labelling did not agree with plate counts. Therefore, a two-step method was developed in which cF labelling was followed by a lactose-energized efflux assay. The combined assay proved to be a good and rapid indicator for reproduction and acidification capacity of stressed L. lactis. This novel assay has potential for physiological research and dairy applications related to LAB. PMID- 10791761 TI - Resistance of Rhodococcus equi to acid pH. AB - Rhodococcus equi is an important gram-positive intracellular facultative pathogen in foals of less than 3 months of age, that causes suppurative bronchopneumonia, lymphadenitis and/or enteritis. The disease in young foals mainly occurs in spring and summer when weather conditions are favorable for survival and multiplication of the bacteria in the environment. R. equi is widespread in the environment of horsebreeding farms: it has been isolated from the soil of paddocks and from the feces of adult horses and foals. Aerosol infection via dust of paddocks seems to be the major route of foal infections. The molecular mechanisms associated with the pathogenesis are not well understood and little is known about the markers or factors associated with virulence of R. equi. However, the discovery of a large plasmid in virulent strains and its association with virulence in mice and in young foals was reported. In this report, we studied the acid resistance of virulent R. equi in comparison with its avirulent plasmid-free isogene. PMID- 10791762 TI - Bibliography of food microbiology. PMID- 10791763 TI - Introduction: IGFs and IGFBPs in the normal mammary gland and in breast cancer. PMID- 10791764 TI - IGF-I: an essential factor in terminal end bud formation and ductal morphogenesis. AB - Growth hormone (GH) is essential for rodent mammary gland development during puberty. It binds to GH receptors in the stromal compartment of the mammary gland and stimulates IGF-I mRNA expression. These findings lead to the hypothesis that GH acts through locally produced IGF-I, which in turn, causes development of terminal end buds (TEBs), the structures that lead the process of mammary gland development during puberty. Subsequent studies have in large measure proven this hypothesis. They include the observations that mammary development was grossly impaired in female mice deficient in IGF-I (IGF-I(-/-) knockout mice), and treatment of these mice with IGF-I plus estradiol (E2) restored pubertal mammary development while treatment with GH + E2 did not. Thus, the full phenotypic action of GH in mammary gland development is mediated by IGF-I. We have demonstrated one effect of GH on the mammary gland that does not appear to be mediated by the action of IGF-I. GH increased the level of estrogen receptor (ER) mRNA and protein in the nuclei of mammary fat pad cells, but IGF-I did not. In addition to the critical role of the GH/IGF-I axis during pubertal mammary development, other data suggest that IGF-I might also be of importance during pregnancy and lactation. In summary, the earliest phase of pubertal mammary development (formation of TEBs) requires IGF-I or GH in IGF-I sufficient animals. No other hormones have been shown to stimulate formation of TEBs unless GH or IGF I is present. GH-induced IGF-I is of major importance in ductal morphogenesis, and may, in fact, be necessary for later stages of mammary development, as well. PMID- 10791765 TI - IGF and insulin action in the mammary gland: lessons from transgenic and knockout models. AB - Transgenic and knockout mice have become valuable experimental systems with which to study specific molecular events within the mammary gland of an intact animal. These models have provided a wealth of information about the effects of a number of oncogenes and growth factors. This review focuses on results obtained from the application of transgenic and knockout models to determine the roles of insulin and insulin-like growth factors (IGF) in the regulation of mammary gland development, lactation and tumorigenesis. Transgenic models which overexpress IGF I or -II display specific alterations in mammary gland development and an increased incidence of mammary tumors. Analysis of mammary gland development in knockout mice which are deficient in IGF-I or the IGF-I receptor supports the conclusion that the IGF system is important for normal mammary gland development. This review discusses these observations in detail and attempts to fit them into a larger picture of IGF and insulin action in the mammary gland. PMID- 10791767 TI - Local IGF-I axis in peripubertal ruminant mammary development. AB - The regulation of mammary growth and development in heifers is accomplished by complex interactions of hormones, growth factors, and extracellular matrix molecules. Many of these growth stimulators are believed to be locally produced in the mammary gland and to be affected by developmental and nutritional status. Although estrogen and growth hormone are considered critical to pubertal mammogenesis, results summarized in this review suggest that IGF-I and IGF binding proteins are especially important locally-produced growth regulators in peripubertal ruminants. This assertion is supported by studies of ovariectomized heifers, in which increased stromal IGFBP-3 and reduced IGF-I correspond with a failure of udder development. Similarly, reduced mammary development with overfeeding coincides with reduced mitogenic activity of mammary tissue extracts and altered concentrations of IGF-I and IGFBPs. In vitro studies convincingly demonstrate that much of the mitogenic activity of mammary extracts or serum can be attributed to IGF-I and that alterations in IGFBP-3 modulate its effectiveness. Thus by analogy to second messenger mechanisms of action for protein hormones, local mammary-derived growth factors likely explain many of the effects attributed to the classic mammogenic hormones. PMID- 10791766 TI - The insulin-like growth factors (IGFs) and IGF binding proteins in postnatal development of murine mammary glands. AB - The insulin-like growth factors are mitogens and survival factors for normal mammary epithelial cells in vitro. Data reviewed here demonstrate that mRNAs for IGF-I and IGF-II, the IGF type I receptor and the IGFBPs are expressed locally in mammary tissue during pubertal and pregnancy-induced growth and differentiation of murine mammary glands. IGF-I, IGF-II and the IGF-IR were expressed in terminal end buds (TEBs) in virgin glands during ductal growth. In addition, IGF-II and IGF-IR mRNAs were expressed in ductal and alveolar epithelium in glands throughout postnatal development. Consistent with these results, IGF-I promoted ductal growth and proliferation in mouse mammary glands in organ culture. In addition to endogenous expression of the IGFs and IGF-IR, the IGFBPs showed a varied pattern of expression in mammary tissue during postnatal development. For example, IGFBP-3 and -5 mRNAs were expressed in TEBs and ducts while IGFBP-2 and 4 mRNAs were expressed in stromal cells immediately surrounding the epithelium. These results support a role for the IGFs and IGFBPs as local mediators of postnatal mammary gland growth and differentiation. PMID- 10791768 TI - Insulin-like growth factor (IGF) system in the bovine mammary gland and milk. AB - Primary bovine mammary cells express the two IGF receptors (IGF-IR, IGF-IIR), insulin receptor, and four IGFBPs (IGFBP-2, -3, -4, and -5). Examination of the IGF-IR during the mammary gland lactation cycle shows that IGF-IR number declines at parturition, a change that coincides with decreases in the blood level of its ligand, IGF-I. IGF-II and IGF-IIR are largely unchanged. IGFBP-3 is the predominant mammary IGFBP and its concentration also declines in blood and milk during lactation compared to prepartum and involution periods. Time of lactation and pregnancy were the main determinants of milk but not blood IGFBP-3 levels. IGFBP-3 binds to membrane proteins of bovine mammary tissue; an IGFBP-3 binding protein has been identified as bovine lactoferrin. Lactoferrin has the capacity to compete with IGF binding to IGFBP-3. Appearance of both IGFBP-3 and lactoferrin in conditioned media of primary cultures of bovine mammary cells was stimulated by all trans retinoic acid (atRA). Furthermore, atRA was necessary for the entry of exogenously added lactoferrin into the mammary cell nucleus, while IGFBP-3 entry into the nuclei of atRA treated cells required the presence of lactoferrin. These findings reveal a novel role for lactoferrin, suggesting that lactoferrin is critically involved in the regulation of the IGF system during the involution period. PMID- 10791769 TI - Insulin-like growth factor binding proteins: IGF-dependent and -independent effects in the mammary gland. AB - The insulin-like growth factor binding proteins (IGFBPs) are a family of proteins which bind to the IGFs with high affinity. Their expression within the mammary gland is species specific; it has thus been difficult to determine the biological roles of these binding proteins during lactation. In this article we propose a role for IGFBP-5 in the mammary gland involving the initiation of apoptosis induced by sequestration of IGF-1, an important survival factor for the mammary gland. We have shown that this binding protein retains its high affinity for IGF 1 and that it is present in extremely high concentrations compared with the growth factor. These observations make it likely that IGFBP-5 is capable of preventing interaction of IGF-1 with its receptor on the epithelial cells synthesizing milk. We have also demonstrated that IGFBP-5 interacts with alpha(s2)-casein and that this interaction implicates it in the regulation of plasminogen activation in the mammary gland. The generation of plasmin is a key initiating event in the remodeling of the extracellular matrix during mammary involution. As such, IGFBP-5 may play a key role in coordinating cell death and tissue remodeling processes. Many of the molecules involved in embryological development are also expressed in the developing and involuting mammary gland. We believe that our studies may offer mechanistic explanations for apoptotic events in a wide variety of tissues. We have recently shown that IGFBP-5 is apoptotic in the chick embryonic limb bud, adding further support to our belief that IGFBP-5 serves this function in the mammary gland. We hope to be able to explore the role of this binding protein in the mammary gland with a transgenic mouse model expressing IGFBP-5 on the beta-lactoglobulin promoter. PMID- 10791770 TI - Insulin-like growth factor binding proteins (IGFBPs) in breast cancer. AB - The IGFBP family comprises six proteins with high affinity for the IGFs and several lower affinity IGFBP-related proteins. Their production in the breast is controlled by hormones, other local regulators and in tumors relates to the estrogen receptor status. Their functional activity can also be affected by various post-translational modifications. The IGFs are generally present at levels far in excess of that required for maximal receptor stimulation, and the IGFBPs are critical regulators of cellular action. IGFBPs can affect cell function in an IGF-dependent or independent manner. IGF bioactivity locally in the breast is influenced not only by local tissue expression and regulation of IGFs, IGFBPs, and IGFBP proteases, but also by these factors delivered from the circulation. Changes in the balance of the components of the IGF system may lead to a disruption of tissue homeostasis. PMID- 10791771 TI - Mannose 6-phosphate/insulin-like growth factor 2 receptor, a bona fide tumor suppressor gene or just a promising candidate? AB - The mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R) is considered a "candidate" tumor suppressor gene. This hypothesis has been provoked by the identification of loss of heterozygosity (LOH) at the M6P/IGF2R locus on chromosome 6q26 in breast and liver cancer, accompanied by point mutations in the remaining allele. Somatic mutations in coding region microsatellites have also been described in replication error positive (RER+) tumors of the gastrointestinal tract, endometrium and brain. These genetic data are compelling, but a tumor suppressor gene candidate has to meet functional as well as genetic criteria. This review weighs the evidence and discusses the observations that are necessary to promote M6P/IGF2R from candidate to bona fide tumor suppressor gene. PMID- 10791772 TI - Function of the IGF-I receptor in breast cancer. AB - The insulin-like growth factor-I receptor (IGF-IR) is a transmembrane tyrosine kinase regulating various biological processes such as proliferation, survival, transformation, differentiation, cell-cell and cell-substrate interactions. Different signaling pathways may underlie these pleiotropic effects. The specific pathways engaged depend on the number of activated IGF-IRs, availability of intracellular signal transducers, the action of negative regulators, and is influenced by extracellular modulators. Experimental and clinical data implicate the IGF-IR in breast cancer etiology. There is strong evidence linking hyperactivation of the IGF-IR with the early stages of breast cancer. In primary breast tumors, the IGF-IR is overexpressed and hyperphosphorylated, which correlates with radio-resistance and tumor recurrence. In vitro, the IGF-IR is often required for mitogenesis and transformation, and its overexpression or activation counteract effects of various pro-apoptotic treatments. In hormone responsive breast cancer cells, IGF-IR function is strongly linked with estrogen receptor (ER) action. The IGF-IR and the ER are co-expressed in breast tumors. Moreover, estrogens stimulate the expression of the IGF-IR and its major signaling substrate IRS-1, while antiestrogens downregulate IGF-IR signaling, mainly by decreasing IRS-1 expression and function. On the other hand, overexpression of IRS-1 promotes estrogen-independence for growth and transformation. In ER-negative breast cancer cells, usually displaying a more aggressive phenotype, the levels of the IGF-IR and IRS-1 are often low and IGF is not mitogenic, yet the IGF-IR is still required for metastatic spread. Consequently, IGF-IR function in the late stages of breast cancer remains one of the most important questions to be addressed before rational anti-IGF-IR therapies are developed. PMID- 10791774 TI - Adjunct cultures: recent developments and potential significance to the cheese industry. AB - Several previous reviews have described different ways to enhance the flavor and texture of cheese, including use of live cells and nonviable attenuated cells as adjunct cultures. However, comparisons between viable and nonviable cultures were never discussed in these reviews. In addition, recent publications on adjunct cultures have not been covered in previous reviews. This article will survey the more recent work on adjunct cultures--with particular attention to whether the adjuncts contained viable or nonviable cells--and propose areas where additional research is needed. PMID- 10791775 TI - Characterization of mesophilic mixed starter cultures used for the manufacture of aged cheddar cheese. AB - Seventy-one different Lactococcus lactis subsp. cremoris strains were isolated from seven mesophilic mixed starters used in the manufacture of aged Cheddar cheese in Canada. Based on plasmid profiles and growth in milk (with or without glucose, Casamino Acids or both), two mixed starters were highly heterogeneous, containing at least 18 to 24 distinct L. lactis strains. Three mixed starters were comprised of seven to nine strains, whereas two starters were relatively homogeneous, containing two or three strains. Many strains with similar plasmid profiles behaved differently during growth in milk, indicating variability in the phenotypes. Only 20% of the strains could grow in plain milk, whereas 30% could not grow in milk supplemented with glucose and Casamino Acids. Twenty-five lactococcal bacteriophages were also isolated from whey samples with single strains as hosts. Eighteen phages belonged to the 936 species and seven to the c2 species. Thirteen strains were insensitive to all 25 phages. Almost all sensitive strains were phage species-specific. The 936-like phages had a broader host range. PMID- 10791773 TI - Crosstalk between the insulin-like growth factors and estrogens in breast cancer. AB - Once it was recognized that breast tumor growth was stimulated by estrogens, successful therapeutic strategies based on depriving the tumor of this hormone were developed. Since the growth stimulatory properties of the estrogens are governed by the estrogen receptor (ER), understanding the mechanisms that activate ER are highly relevant. In addition to estrogens, peptide growth factors can also activate the ER. The insulin-like growth factors (IGFs) are potent mitogens for ER-positive breast cancer cell lines. This review will examine the evidence for interaction between these two pathways. The IGFs can activate the ER, while ER transcriptionally regulates genes required for IGF action. Moreover, blockade of ER function can inhibit IGF-mediated mitogenesis and interruption of IGF action can similarly inhibit estrogenic stimulation of breast cancer cells. Taken together, these observations suggest that the two growth regulatory pathways are tightly linked and that a further understanding of the mechanism of this crosstalk could lead to new therapeutic strategies in breast cancer. PMID- 10791776 TI - Comparison of newly isolated strains of Lactobacillus delbrueckii subsp. lactis for hydrogen peroxide production at 5 degrees C. AB - Isolates of Lactobacillus delbrueckii subsp. lactis obtained from raw milk samples were compared for the ability to produce hydrogen peroxide (H2O2) at 5 degrees C. Nineteen out of 101 lactobacilli isolated were identified as L. delbrueckii subsp. lactis. The isolates of L. delbrueckii subsp. lactis from most raw milk samples produced more H2O2 than did isolates of other species of lactobacilli from the same samples. Seven isolates of L. delbrueckii subsp. lactis, which produced the highest levels of H2O2 at 5 degrees C were selected for comparison with a laboratory strain, L. delbrueckii subsp. lactis I. In 24 h, isolate RM2-5 produced 7.0 microg/10(9) cfu in buffer containing 5 mM sodium lactate and 4.4 microg/10(9) cfu in buffer containing 5 mM glucose. Three other isolates also produced more H2O2 on sodium lactate than on glucose. However, three remaining new isolates produced more H2O2 on glucose than on sodium lactate. All seven of the most active new isolates of L. delbrueckii subsp. lactis produced significantly higher concentrations of H2O2 than did L. delbrueckii subsp. lactis I in both solutions. Strain RM2-5 produced more H2O2 than did the other six most active newly isolated strains of L. delbrueckii subsp. lactis in this comparison. PMID- 10791777 TI - Growth associated exopolysaccharide expression in Lactococcus lactis subspecies cremoris Ropy352. AB - A natural lactococcal isolate, Lactococcus lactis ssp. cremoris Ropy352, has been previously shown to express two phenotypically distinct exopolysaccharides (ropy and mucoid). This natural isolate was cultured on various media to explore the carbon requirements for exopolysaccharide expression. Ropy exopolysaccharide expression was optimal when grown in defined media rather than on M17-based media. Ropy352 was examined for inducible lysogenic phages. No lytic burst was observed in Ropy352 with ultraviolet light or mitomycin C for phage induction. The sugar compositions of the two phenotypically distinct exopolysaccharides were determined. The ropy exopolysaccharide is composed of galactose and glucose in the molar percents of 42 and 58%, respectively. The mucoid exopolysaccharide is composed of galactose, glucose, and mannose in the molar percents of 58, 29, and 13%, respectively. Mutational analysis revealed that mutations impairing ropy exopolysaccharide expression but not affecting mucoid exopolysaccharide expression could be isolated. PMID- 10791778 TI - Preliminary observations on influence of dairy products on biofilm removal from silicone rubber voice prostheses in vitro. AB - We determined oropharyngeal biofilm removal from silicone rubber voice prostheses in an artificial throat after perfusion with different commercially available dairy products, including buttermilk, Lactobacillus casei Shirota fermented milk (Yakult, Yakult Netherlands BV, Almere, The Netherlands), low-fat milk, and three types of yogurt. Buttermilk removed both yeasts and bacteria from biofilms on voice prostheses, but this was not observed with a pasteurized buttermilk product. Some of the other products, most notably Yakult fermented milk drink, reduced the number of bacteria, whereas growth of yeasts in the biofilms was not inhibited. This study demonstrates that there may be health benefits associated with the consumption of certain dairy products when applied to the upper digestive tract. PMID- 10791779 TI - Effect of milk preacidification on low fat mozzarella cheese. I. Composition and yield. AB - The effect of milk preacidification with acetic or citric acid on the composition and yield of low fat Mozzarella cheese was determined. Two cheese manufacturing trials were conducted. In trial 1, three vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control) and preacidification to pH 6.0 and pH 5.8 with citric acid. In trial 2, four vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. Cheese manufacture was repeated on three different days in trial 1 and four different days in trial 2 using a randomized-complete block design. Preacidification to pH 5.8 with citric acid decreased cheese calcium more than preacidification to pH 5.8 with acetic acid. Preacidification with citric acid in trial 1 decreased protein recovery in the cheese, and there was a trend for decreased protein recovery in the cheese for trial 2. Differences in fat recovery due to preacidification varied, sometimes being lower than the control other times being higher than the control. The reduction in calcium and protein recovery in the cheese caused by preacidification lowered composition adjusted cheese yield and yield efficiency. Yield efficiency was reduced by about 2.5 and 5.5%, respectively, with preacidification to pH 6.0 and 5.8 with citric acid. Yield efficiency was reduced by about 2.2 and 3.4%, respectively, with preacidification to pH 6.0 and pH 5.8 with acetic acid. PMID- 10791780 TI - Pyroglutamic acid in cheese: presence, origin, and correlation with ripening time of Grana Padano cheese. AB - Pyroglutamic acid is present in many cheese varieties and particularly in high amounts (0.5 g/100 g of cheese) in extensively ripened Italian cheeses (Grana Padano and Parmigiano Reggiano) that are produced with thermophilic lactic acid bacteria as starters. The mechanism of pyroglutamic acid formation in cheese seems to be mostly enzymatic, as demonstrated by the presence of only L pyroglutamic acid enantiomer. Thermophilic lactobacilli are involved in pyroglutamic acid production, as suggested by the low pyroglutamic acid content found in Bagos, a ripened Italian mountain cheese produced without addition of starter. Because milk pasteurization did not influence the pyroglutamic acid content in the ripened Grana Padano cheese, the formation of pyroglutamic acid mainly depends on the whey starter microflora rather than that of raw milk. Pyroglutamic acid concentration is linearly correlated (R2 = 0.94) with the age of Grana Padano cheese. PMID- 10791781 TI - Probiotic culture survival and implications in fermented frozen yogurt characteristics. AB - Low-fat ice cream mix was fermented with probiotic-supplemented and traditional starter culture systems and evaluated for culture survival, composition, and sensory characteristics of frozen product. Fermentations were stopped when the titratable acidity reached 0.15% greater than the initial titratable acidity (end point 1) or when the pH reached 5.6 (end point 2). Mix was frozen and stored for 11 wk at -20 degrees C. The traditional yogurt culture system contained the strains Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. The probiotic-supplemented system contained the traditional cultures as well as Bifidobacterium longum and Lactobacillus acidophilus. We compared recovery of Bifodobacterium by three methods, a repair-detection system with roll-tubes and plates on modified bifid glucose medium and plates with maltose + galactose reinforced clostridial medium. Culture bacteria in both systems did not decrease in the yogurt during frozen storage. The roll-tube method with modified bifid glucose agar and repair detection system provided at least one-half log10 cfu/ml higher recovery of B. longum compared with recoveries using modified bifid glucose agar or maltose + galactose reinforced clostridial agar on petri plates. No change in concentrations of lactose or protein for products fermented with either culture system occurred during storage. Acid flavor was more intense when product was fermented to pH 5.6, but yogurt flavor was not intensified. The presence of probiotic bacteria in the supplemented system seemed to cause no differences in protein and lactose concentration and sensory characteristics. PMID- 10791782 TI - High pressure inactivation of microorganisms inoculated into ovine milk of different fat contents. AB - High hydrostatic pressure inactivation of Escherichia coli, Pseudomonas fluorescens, Listeria innocua, Staphylococcus aureus, and Lactobacillus helveticus were studied. These microorganisms were inoculated at a concentration between 10(7) and 10(8) cfu/ml in Ringer solution and in ovine milk adjusted to 0, 6, and 50% fat content to evaluate the baroprotective effect of fat content on inactivation of microorganisms. Treatments of pressurization consisted of combinations of pressure (100 to 500 MPa) and temperature (4, 25, and 50 degrees C) for 15 min. Gram-negative microorganisms were more sensitive than were Gram positive ones (more destruction P. fluorescens > E. coli > or = List. innocua > Lb. helveticus > S. aureus). Pressurizations at low temperature (4 degrees C) produced greater inactivation on P. fluorescens, List. innocua, and Lb. helveticus than at room temperature (25 degrees C), whereas for E. coli and S. aureus the results were opposite. Ovine milk per se (0% fat) showed a baroprotective effect on all microorganisms, but percentage of fat (6 and 50%) did not show a progressive baroprotective effect in all pressurization conditions or for all microorganisms. PMID- 10791783 TI - Presence of a glycan at a potential N-glycosylation site, Asn-281, of bovine lactoferrin. AB - This work was performed to clarify the differences in glycan moieties between multiple molecular mass forms of bovine lactoferrins (bovine lactoferrins-a and b). After digestion of both bovine lactoferrins with cyanogen bromide and V8 protease, glycopeptides were successively purified by concavalin A affinity chromatography and HPLC on an octadecylsilyl column. Four glycopeptides glycosylated at Asn-233, -368, -476, and -545 were obtained from both hydrolysates of bovine lactoferrins-a and -b. On the other hand, a glycopeptide glycosylated at Asn-281 was only detected in hydrolysate of bovine lactoferrin-a, indicating that bovine lactoferrin-a possessed five N-glycosylated sites. The glycan linked to Asn-281 of bovine lactoferrin-a was found to consist of fucose, galactose, and N-acetylgalactosamine in addition to mannose and N acetylglucosamine. HPLC analysis of this glycan on a normal phase column showed that peaks of several glycans were detected. These glycans changed to one major glycan consisting of only mannose and N-acetylglucosamine on exoglycosidase digestion. From these results, this glycan seemed to be of the complex type and possess heterogeneous structure. PMID- 10791784 TI - Effects of different forms of canola oil fatty acids plus canola meal on milk composition and physical properties of butter. AB - Twenty multiparous Holstein cows were used in a 16-wk trial. A block of 10 cows received a control diet, based on corn silage, and the other block of 10 cows successively received four diets with 1) an extruded blend of canola meal and canola seeds, 2) canola meal and whole canola seeds, 3) canola meal and ground canola seeds, or 4) canola meal and calcium salts of canola oil fatty acids. Canola fat represented about 2% of dietary dry matter. Compared to control cows, treated cows had similar dry matter intake, milk production, and daily milk output of true protein or fat. Protein contents of milk was decreased by all treatments, with a lower effect of extruded or whole canola seeds. Milk fat contents was lowered by all treatments, extruded seeds and calcium salts resulting in most important effects. All treatments lowered the percentage of fatty acids with 12 to 16 carbons in milk fat, increased C18:0 and cis-C18:1 percentages, and the proportion of liquid fat in butter between 0 and 12 degrees C. Calcium salts and, to a lesser extent extruded seeds, resulted in most important improvements of milk fatty acid profile and butter softness, whereas whole seeds had low effects. PMID- 10791785 TI - Hock lesions and free-stall design. AB - We compared the prevalence and severity of skin lesions on the hocks of lactating dairy cows in southern British Columbia, comparing 20 farms using three common bedding surfaces: sawdust, sand, and geotextile mattresses. Skin lesions were scored at five positions on the hock. For each position we noted if the lesion showed inflammatory attributes, and then assigned a severity score. Of the 1752 lactating cows scored, 1267 cows (73%) had at least one hock lesion. Of those cows with lesions, 87% had lesions on both legs, 76% had lesions on more than one location on the hock, and 78% had a lesion of at least moderate severity (i.e., evidence of skin breakage or an area of hair loss >10 cm2). Lesions were most prevalent on farms that used geotextile mattresses (91% of cows) and least common on farms that used sand (24% of cows). Moreover, lesions on cows from farms using mattresses were more numerous and more severe than those on cows from sand-bedded farms. The prevalence and severity of lesions on farms using sawdust was intermediate. Lesions also varied in relation to location on the hock. For farms using geotextile mattresses, lesions were more common and more severe on the lateral surfaces of both the tuber calcis and the tarsal joint. On farms using sawdust, lesions were common on the dorsal surface of the tuber calcis and the lateral surfaces of both the tuber calcis and the tarsal joint. Lesions were rare on all five positions for cows from sand-bedded farms. Among the 10 farms sampled using sawdust, we found a significant negative relationship between the length of the stall and severity of lesions. For cows with lesions, the number and severity of lesions increased with age. PMID- 10791786 TI - Impact of dietary protein amount and rumen undegradability on intake, peripartum liver triglyceride, plasma metabolites, and milk production in transition dairy cattle. AB - Feeding strategies of transition dairy cows contribute to the risk factors associated with metabolic disorders that limit production in the ensuing lactation. To investigate the effects of prepartum dietary crude protein (CP) concentration and amount of rumen-undegradable protein (RUP) on postpartum health and production, 44 multiparous Holstein cows were blocked by expected calving date and assigned to one of four isoenergetic prepartum rations beginning 28 d prior to expected calving date. Prepartum rations were: 12% CP and 26% RUP, 16% CP and 26% RUP, 16% CP and 33% RUP, or 16% CP and 40% RUP on a dry matter basis. All cows were fed the same postpartum diet (18% CP, 40% RUP) from 1 to 56 d in milk (DIM). Prepartum dry matter intake (DMI) was not different among dietary treatments. Mean postpartum intakes (kg/d) were higher through 56 DIM (P<0.05) for cows fed the 12% CP:26% RUP diet prepartum compared with any of the 16% CP diets (21.8 vs. 19.8, 18.6 and 18.6; 12% CP:26% RUP vs. 16% CP:26% RUP, 16% CP:33% RUP and 16% CP:40% RUP). There was a DIM x prepartum diet interaction (P<0.05) with the greatest effect of the 12% CP:26% RUP diet evident during the first 35 DIM. Cows fed the 12% CP:26% diet during the transition period tended to produce more milk (kg/d) (P = 0.08) than did cows fed any of the 16% CP diets (40.8 versus 37.8, 38.7, and 37.4; 12% CP:26% RUP vs. 16% CP:26% RUP, 16% CP:33% RUP, and 16% CP:40% RUP). Additional protein (12 vs. 16% CP) in the prepartum diet tended to decrease milk protein (P = 0.10) and milk fat yield (P = 0.08) but did not alter percent milk fat, percent milk protein, or MUN. Liver triglyceride (TG) expressed as milligrams of TG per microgram of DNA or percentage of dry matter (DM) on d -28, -14, +1, +28, and +56 relative to calving were not significantly different among treatments. Maximal (P<0.05) infiltration of TG in liver was observed on +1 d when expressed as a percentage of DM and on +28 d when expressed as milligrams of TG per microgram of DNA. Plasma glucose, calcium, urea nitrogen, beta-hydroxybutyrate, and nonesterified fatty acids were not different (P<0.05) among treatments. The data indicate carryover effects of prepartum dietary protein on postpartum intake and milk production, pointing to beneficial effects of maintaining dietary protein for dairy cows in late gestation at 12% CP. PMID- 10791787 TI - A computerized mastitis decision aid using farm-based records: an artificial neural network approach. AB - A computer module was developed and tested that used field survey and Dairy Herd Improvement Association (DHIA) data to broadly classify bacterial causes of mastitis in dairy herds. Further development of the computer model could aid interpretation of DHIA data by dairy record processing centers and herd consultants. This diagnostic module was developed with an artificial neural network, a technology that processes complex data in a manner similar to human brain function. Information describing herd management practices, quarter milk samples, and monthly DHIA data was collected from Pennsylvania dairy herds with moderate to high somatic cell counts. This information was used to develop or train an artificial neural network model that discriminated among four categories of bacterial organisms (contagious, environmental, no significant growth, and other) associated with clinical and subclinical mastitis. After training the model, new DHIA and management data were presented to the model to assess its ability to classify bacteriological etiology. When the artificial neural network was used, the probabilities of diagnosing the bacteriologic status from three randomly selected cow groups and from new untested herds ranged from 57 to 71%. Performance of the artificial neural network model was best in herds with higher frequency of minor and contagious pathogens. Prediction results for the same test data with linear discriminant analysis were less successful, ranging from 42 to 57%. PMID- 10791788 TI - A stochastic model simulating the feeding-health-production complex in a dairy herd. AB - A dynamic, stochastic, and mechanistic Monte Carlo model, simulating a dairy herd with focus on the feeding-health-production complex is presented. By specifying biological parameters at cow level and a management strategy at herd level, the model can simulate the technical and economic consequences of scenarios at herd level. The representation of the feeding-health-production complex is aimed to be sufficiently detailed, to include relationships likely to cause significant herd effects, and to be sufficiently simple to enable a feasible parameterization of the model and interpretation of the results from the model. Consequently, diseases are defined as four disease types: two metabolic disease types, an udder disease type, and a reproductive disease type. Risk factors for the diseases were defined as parity, yield capacity, disease recurrence, disease interrelationships, lactation stage, and season. Direct effects of the diseases were defined according to milk yield, feed intake, feed utilization, conception, culling, involuntary removal, and death. Scenarios differing in base risks of milk fever and ketosis, heat detection rate, and culling strategy were simulated for describing the model behavior. Annual milk yield per cow was decreased by increased risk of ketosis and by increased risk of milk fever, even though no direct effect of milk fever on milk yield was modeled at the cow level. The indirect effect from milk fever is a consequence of increased replacement rate (relatively lower milk yield from younger cows). By ignoring the history of milk fever in insemination and replacement decisions, a significantly reduced net income per cow was found in some herds. We concluded that important benefits from using such a herd model are the capability of accounting for herd management factors and the advantage of avoiding to double count the indirect effects from disease, such as increased risk of other diseases, poorer reproduction results, and increased risk of culling and death. PMID- 10791789 TI - Plasma concentration of urea, ammonia, glutamine around calving, and the relation of hepatic triglyceride, to plasma ammonia removal and blood acid-base balance. AB - Two experiments were conducted to test the following two hypotheses: 1) fatty liver could hamper hepatic conversion of ammonia to urea and increase circulating ammonia or Gln% [Gln% = Gln x 100/(Gln + Glu)] in cows around parturition; 2) decreased ureagenesis might cause alkalosis and in turn reduce blood Ca. In the first experiment, 14 Holstein cows were monitored from 27 d prepartum to 35 d postpartum. There was a rise in circulating ammonia and Gln% at calving, suggesting an increase in ammonia passing to and through the liver. Stepwise regression analysis revealed the following relationship for plasma samples at 22 h and liver triglyceride at 2 d postpartum: ammonia (microM) = 32.1+/-0.89 triglyceride (% DM), Gln% = 71.2 + 0.23 triglyceride (% DM) + 1.31 urea (mM). The positive correlation between liver triglyceride and plasma ammonia and Gln% suggests that hepatic triglyceride accumulation might inhibit ureagenesis, thereby increasing ammonia concentration at the perivenous hepatocytes where Gln synthesis occurs and increasing ammonia concentration in blood leaving the liver. In the second experiment, 28 rats were used to determine whether hepatic triglyceride accumulation, induced by choline deficiency, affects urinary ammonia N and blood pH homeostasis. There was a trend for a positive correlation between urinary ammonia N and liver triglyceride. No correlation between liver triglyceride and blood pH, bicarbonate, pCO2 or plasma Ca was found. In conclusion, hepatic triglyceride accumulation may inhibit ureagenesis and result in increased circulating ammonia, Gln% and urinary ammonia N in vivo. Hepatic triglyceride accumulation did not affect blood pH homeostasis. PMID- 10791791 TI - Sites of digestion and bacterial protein synthesis in dairy heifers fed fresh oats with or without corn or barley grain. AB - Six Holstein-Friesian heifers fitted with ruminal, duodenal, and ileal cannulas were used in a replicated 3x3 Latin square to study the effects of partial replacement [1:1 dry matter (DM) basis] of fresh winter oats (WO) by ground corn (C) or barley (B) on digestion and bacterial protein synthesis. Supplemented diets contained 24% starch, and all diets were fed indoors at 2.5% of body weight (DM basis). Ruminal and total tract digestibilities of organic matter and neutral detergent fiber were similar for all treatments. Ruminal and total tract starch digestibility was similar for C and B diets. Nitrogen intake was greater for WO than for supplemented diets. However, duodenal flows of nonammonia N and bacterial N did not differ among treatments. The efficiency of bacterial protein synthesis was similar for the three diets, suggesting that the fermentation of high quality fresh forage DM provided sufficient energy for the microorganisms in the rumen. Ruminal ammonia-N concentration was greater for WO than for supplemented diets, and for C than for B diet. Ruminal fluid pH and concentrations of total volatile fatty acid were not different among diets, but concentration of acetate was higher and that of propionate was lower for WO than for supplemented diets and for C than for B diet. Supplementation of WO with barley rather than with corn decreased C2:C3 ratio without affecting fiber digestion. Supplements increased N utilization relative to N intake but did not increase duodenal nonammonia N flow. PMID- 10791790 TI - Efficacy of two modified nonantibiotic formulations (Victory) for treatment of papillomatous digital dermatitis in dairy cows. AB - A field trial was conducted to compare the efficacy of the original and two modified formulations of Victory and oxytetracycline among dairy cows affected with papillomatous digital dermatitis. Seventy-eight cows with papillomatous digital dermatitis lesions were randomly allocated to one of four treatment groups (A, B, C, D). Cows in group A (n = 19) were treated with an oxytetracycline solution; cows in group B (n = 22) were treated with the original formulation of Victory containing soluble copper, peroxide compound, and a cationic agent; cows in group C (n = 17) were treated with a modified formulation of Victory containing reduced soluble copper and peroxide compound but increased levels of cationic agent; and cows in group D (n = 20) were treated with a modified formulation of Victory containing levels of soluble copper and cationic agent equivalent to the original formulation but with reduced concentrations of peroxide compound. Cows were examined 7, 14 and 28 d after initial treatment; during each examination, pain and lesion scores were recorded. The modified nonantibiotic formulation used in cows in group C appeared to be the most effective for treatment of papillomatous digital dermatitis. Proportions of cows with signs of pain were significantly lower among cows in group C, compared with cows in group A. Similarly, pain scores were significantly lower among cows in treatment group C, compared to cows in group A. The 2 low efficacy of oxytetracycline was an unexpected result b and may have clinical implications associated with possible antibiotic resistance in dairy cows affected with papillomatous digital dermatitis. PMID- 10791792 TI - Rumen fermentation in lactating cows selected for milk fat content fed two forage to concentrate ratios with hay or silage. AB - Sixteen multiparous cows, including eight rumen fistulated cows, were used in a 4x4 Latin square experiment designed to study dietary effects on rumen and blood parameters and milk production in cows differing in genetic capacity for milk fat content. Diets contained forage to concentrate ratios of 50:50 or 30:70 with either grass hay or silage as the forage. Ruminal fermentation was characterized by a high molar percentage of butyrate, 14 to 17%. Forage to concentrate ratio affected most rumen parameters, with the exception of the molar percentage of propionate (18 to 19%). The silage had a higher fiber degradation rate compared with hay. Compared to hay diets, silage diets had higher ruminal outflow rates, lower acetate:propionate ratios, and greater milk production with no differences in milk composition. Cows selected for low milk fat had higher molar percentages of propionate in the rumen. The low milk fat cows had higher milk production than cows selected for high milk fat but did not differ in milk fat yield. Cows fed the 30:70 diets had higher plasma insulin concentrations in response to a glucose challenge. The low milk fat cows had lower basal concentrations of insulin and lower insulin responses to a glucose challenge. Small changes in nutrient metabolism and supply were sufficient to influence milk production. PMID- 10791793 TI - Amino acid exchange by the mammary gland of lactating goats when histidine limits milk production. AB - The aim of this study was to monitor amino acid (AA) exchange kinetics of the mammary gland in response to an imposed limitation on His supply for milk production. Lactating goats (n = 4, approximately 120 DIM) were fed a low protein ration that provided only 77% of metabolizable protein and 100% of energy requirements for milk production. The protein deficiency was alleviated by infusion into the abomasum of an AA mixture (67 g/d) including (+H; 4.4 g/d) or excluding (-H) His. Goats were assigned to treatments (6 to 7 d) according to a switchback design. On the last day of the first two periods, [U-13C]AA were continuously infused i.v. for 7 h and arterial and mammary vein blood was withdrawn to determine plasma AA concentration and enrichment. Flow probes monitored mammary blood flow. The secretion and enrichments of AA in milk casein were monitored each hour. A three-pool model of the gland was used to derive bi directional rates of plasma AA exchange. Arterial plasma His concentration was lower during -H infusion (8 vs. 73 microM), but those of other AA changed little. Responses to low levels of plasma His were: 1) mammary blood flow increased by approximately 33%; 2) the gland's capacity to remove plasma His increased 43 fold, whereas the gland's capacity for other AA declined by two- to threefold; and 3) influx and efflux of His by the gland decreased. Thus, as the reduction in His efflux was insufficient to offset the reduced influx, milk protein yield decreased from 118 to 97 g/d. PMID- 10791794 TI - Effects of Isotricha, Dasytricha, Entodinium, and total fauna on ruminal fermentation and duodenal flow in wethers fed different diets. AB - The objective was to measure rumen fermentation and duodenal flow of amino acids and nonammonia N components in five groups of five ruminally and duodenally cannulated wethers that were fauna-free, or inoculated with the ciliate protozoa genera of Isotricha, Dasytricha, Entodinium, or a normal population (total) of fauna. They were used in two 25-d periods and fed a haycrop-based diet in the first period and a corn silage-based diet in the second period. Feces, duodenal digesta, and rumen contents were sampled in each period and analyzed. The number of Entodinium in wethers containing the Entodinium monofauna was higher than the total protozoa numbers, including Entodinium, in wethers containing total fauna population. The type of diet or fauna did not affect total volatile fatty acid concentrations in rumen fluid. The ammonia N concentration in rumen fluid was higher in wethers containing total fauna (25 mg/100 ml) than in fauna-free wethers fed the two diets (18 and 12 mg/ 100 ml). In comparison with the respective fauna-free wethers, the concentration of ammonia in wethers containing Entodinium was higher when fed the corn silage diet, but not different when fed the haycrop diet. Ruminal presence of total fauna or Entodinium decreased the nonammonia N by 16 and 17%, and total amino acid flows from the stomach by 20 and 19%, respectively. Flow of bacteria N was decreased in wethers fed the two diets when Entodinium or total fauna were present in the rumen. The presence of Isotricha resulted in higher flow of bacteria N in wethers fed the haycrop diet, but the presence of Dasytricha resulted in higher bacteria N flow in wethers fed the corn silage diet. Entodinium was the most detrimental of ciliate protozoa species concerning protein nutrition of the host ruminant. PMID- 10791795 TI - Effects of hydrolyzed spray dried red blood cells in milk replacer on calf intake, body weight gain, and efficiency. AB - An alternative protein ingredient based on spray-dried, hydrolyzed red blood cells was evaluated in calf milk replacers. Two experiments were conducted to determine the value of the ingredient on intake, growth, and feed efficiency in dairy calves. In experiment 1, Holstein bull calves (n = 120) were fed calf milk replacer containing 0, 11, 22, or 43% of crude protein as spray dried hydrolyzed red blood cells. Calves were fed 454 g/d of experimental milk replacer reconstituted to 12% dry matter plus a conventional calf starter for 28 d. Body weight gain, intake of milk replacer and calf starter, feed efficiency, fecal scores, and days scouring were unaffected by source of protein. In experiment 2, Holstein calves (n = 69) at the University of Minnesota, Crookston and Waseca were fed milk replacer containing 0, 22, or 43% of crude protein as spray dried hydrolyzed red blood cells. Calves were fed 454 g/d of experimental milk replacer reconstituted to 12% dry matter plus a conventional calf starter containing 0 or 25% alfalfa meal for 35 d. No calves died during the study. Body weight gain, feed efficiency, intake of calf starter and milk replacer, fecal scores, and days scouring were unaffected by increasing hydrolyzed red blood cells in milk replacer. Similar performance of all calves indicated that spray dried hydrolyzed red blood cells can replace up to 43% of crude protein from whey protein concentrate without detrimental effects on animal performance. PMID- 10791796 TI - Whole genome scan to detect quantitative trait loci for conformation and functional traits in dairy cattle. AB - A granddaughter design was used to locate quantitative trait loci determining conformation and functional traits in dairy cattle. In this granddaughter design, consisting of 20 Holstein Friesian grandsires and 833 sons, genotypes were determined for 277 microsatellite markers covering the whole genome. Breeding values for 27 traits, regarding conformation (18), fertility (2), birth (4), workability (2), and udder health (1), were evaluated in an across-family analysis using multimarker regression. Significance thresholds were determined using a permutation test. The across-family analysis suggested the presence of 61 quantitative trait loci when 27 (i.e., one for each trait) were expected by chance. The test statistic exceeded the genomewise significance threshold for the following traits and chromosomes: chest width on chromosome 2; gestation length on chromosome 4; stature, body capacity, and size on chromosome 5; dairy character on chromosome 6; angularity on chromosome 12; fore udder attachment on chromosome 13; and fore udder attachment and front teat placement on chromosome 19. The quantitative trait loci for size traits on chromosomes 2, 5, and 6 may also have an effect on calving ease. The quantitative trait loci for udder traits on chromosomes 13 and 19 may also affect somatic cell score and mastitis resistance. If there are no negative effects on other economically important traits, marker assisted selection using markers associated with these quantitative trait loci can be applied. PMID- 10791797 TI - Effect of herd environment on the genetic and phenotypic relationships among milk yield, conception rate, and somatic cell score in Holstein cattle. AB - A total of 248,230 primiparous records of Holstein cows calving from 1987 to 1994 (daughters of 588 sires in 3042 herds) was used to evaluate potential genotype by environment interactions among mature equivalent milk yield, lactation mean somatic cell score, and conception rate at first service. Herds were classified into low and high environmental groups using three different criteria: standard deviation of herd mature equivalent milk yield, a combination of herd mature equivalent milk yield mean and standard deviation, and the herd mean of body weight at first calving divided by age at first calving. Genetic parameters were modeled by using multiple-trait linear mixed models and were fitted using the multiple-trait derivative-free software. Heritabilities for mature equivalent milk yield, lactation mean somatic cell score, and conception rate at first service were 0.221, 0.106, and 0.015 in low environment herds and 0.300, 0.093, and 0.009 in high environment herds, respectively. Genetic (and phenotypic) correlations between mature equivalent milk yield and lactation mean somatic cell score, mature equivalent milk yield and conception rate at first service, and lactation mean somatic cell score and conception rate at first service were 0.277, -0.417, and -0.209, (-0.049, -0.180, and -0.040) and 0.173, -0.318, and 0.144, (-0.087, -0.166, and -0.035) in low and high environment herds, respectively. The genetic correlations between pairs of traits were consistently smaller in high environment herds, suggesting that differences in management between the two environment levels lessened the antagonistic genetic association between the traits studied. A long-range plan for low environment herds should focus on improving the level of management, which would greatly reduce the unfavorable correlated changes in lactation mean somatic cell score and conception rate at first service associated with the genetic improvement of mature equivalent milk yield. PMID- 10791798 TI - A multiple-trait herd cluster model for international dairy sire evaluation. AB - International dairy sire evaluations have traditionally been calculated using a two-step process. Lactation records within each country are used to predict national estimated breeding values, then these national breeding values are transformed to the genetic base, scale, and units of measurement of other countries by using conversion formulae or the multiple-trait, across-country evaluation method. A major limitation of this approach is the need to define environments (traits) according to country borders. Herds located in small, neighboring countries may be much more similar in management, climate, and genetic background than herds located far apart within a single large country. In the present study, international genetic evaluation with herd clusters is proposed. Data consisted of 4.6 million lactation records from 46,000 herds in Austria, Belgium, Czech Republic, Denmark, Estonia, Finland, Israel, Switzerland, and five regions of the US (Midwest, Northeast, Northwest, Southeast, and Southwest). Herds were grouped into clusters based on data of 13 descriptive variables: herd size, calving interval, milking frequency, age at first calving, milk yield, month of calving, predicted transmitting ability of sire for milk, percentage North American genes of sire, latitude, altitude, temperature, rainfall, and percentage of arable land used for pasture. Five clusters were formed; each cluster contained herds from 5 to 11 countries or regions. Genetic correlations between herd clusters ranged from 0.81 to 0.97. The herd cluster model is intuitively appealing, because genetic merit of an animal is predicted for each unique environment or management system, regardless of country borders. This model is parsimonious (the number of traits was reduced from 13 to 5) and is computationally feasible for large data sets. PMID- 10791799 TI - Use of computerized mate selection programs to control inbreeding of Holstein and Jersey cattle in the next generation. AB - The expected role of computerized mate selection programs with regard to inbreeding and lifetime profitability of Holstein and Jersey cattle was examined using data from 25 large registered herds of each breed. Sire selection and mate allocation were carried out using linear programming with the following objectives: 1) minimum inbreeding, 2) maximum net merit subject to a fixed inbreeding threshold, and 3) maximum expected lifetime profit after adjustment for inbreeding depression. Inbreeding of actual matings was similar to inbreeding from random matings, indicating that current inbreeding avoidance programs in these herds are ineffective. Inbreeding was reduced by 1.6 and 1.9% in Holsteins and Jerseys, respectively, when a mate allocation program was applied with service sires and usage levels fixed at the actual values. Benefits of mate selection programs increased when both sire selection and mate pair allocation were considered. Maximization of mean net merit with inbreeding restricted to a fixed level (5% in Holsteins and 8% in Jerseys) led to decreases in inbreeding of 0.9 and 1.4% and increases in lifetime profit of $16.66 and $26.86 in Holsteins and Jerseys, respectively, relative to programs that ignored inbreeding. Maximization of mean expected lifetime profit after adjustment for inbreeding depression decreased inbreeding by 1.8 and 2.8% and increased lifetime profit by $37.37 and $59.77 in Holsteins and Jerseys, respectively. Inbreeding coefficients estimated with pedigree traced to 1985 were inadequate predictors of inbreeding coefficients estimated with pedigrees traced to 1960. Mate selection programs cannot function optimally unless extensive historical pedigree data are available, particularly for service sires. Computerized mate selection programs can reduce inbreeding in the next generation, which will lead to an increase in farm profitability. However, if genetic diversity is to be maintained in the long term, procedures for selecting parents of AI sires must also be considered. PMID- 10791800 TI - Bivariate animal model estimates of genetic, phenotypic, and environmental correlations for production, reproduction, and somatic cells in Jerseys. AB - Correlations were obtained between 18 response variables of a Jersey herd (Florida Agricultural Experiment Station, Gainesville) for 374 first lactations. Estimates were from the use of multivariate, derivative-free, restricted maximum likelihood procedures with the simplex method of partial maximization. Estimates agreed closely with those obtained previously by other methods in this and other dairy populations. All correlations between yields were high and positive; those between yields and days from parturition to first service were negative and near zero. Correlations between yields and somatic cell scores were moderate and negative; those between yields and constituent percentages in general were negative, except for the yield and percentage of the same constituent. Genetic correlations between chloride content and somatic cells and between measures of somatic cells were 1.0. Results suggest that single-trait selection for milk yield should result in correlated increases in constituent yields with slight decreases in percentage composition of constituents and somatic cell counts. PMID- 10791801 TI - Differentiated dairy grazing intensity in the Northeast. AB - As the dairy industry in the Northeast experienced difficult economic conditions in the 1990s, grazing was increasingly viewed as an option for feeding dairy cattle. This analysis used a large sample of dairy farms randomly drawn from three states (Pennsylvania, Vermont, and Virginia) in early 1997 to compare important aspects of the farming operations for four distinct grazing systems: continuous, traditional, moderately intensive, and intensive. Farmers who used intensive grazing tended to be younger, have more cows per acre, and have greater satisfaction with their farming operations. Logit regression results showed that more formal education and a higher debt-to-asset ratio increased the likelihood that a farmer would increase reliance on grazing in the future. PMID- 10791802 TI - Characteristics of veal calves upon arrival, at 28 and 84 days, and at end of the production cycle. AB - Holstein bull calves arriving at five special-fed veal farms (eight production groups) were scored for various physical condition traits and blood-sampled within 2 h after arrival and at 28 d, 84 d, and 1 wk prior to slaughter at 116 to 143 d. Of 1179 total calves in the production groups, 758 calves were scored and evaluated. Blood cell analyses (red and white blood cell counts, hemoglobin, and packed cell volume) were conducted at all four sampling times; total serum protein concentration was obtained at 0 and 28 d. The study was initiated in autumn and ended the following autumn. Mean initial and final body weights were 46.3+/-0.17 and 209.7+/-0.77 kg; mean mortality was 2.5%. Average daily gain of the eight groups ranged from 1.23 to 1.70 kg/d. Subjective scores of 5 = excellent to 1 = very poor condition were used to evaluate 16 different physical condition characteristics. With the exception of leg joint, hoof, and foot scores, most of the physical condition scores exhibited improvement during the first 28 d. Foot and leg impairments did not appear to hinder the ambulatory ability of the calves during the production period. Physical condition scores at d 0 and 28 were generally not related to numbers or types of medical treatments (enteric, respiratory, other, or total) or to average daily gain during the production period. Means for most erythrocytic and leukocytic traits upon arrival (d 0) were within normal ranges, although 27.4% of the calves were clinically or marginally anemic. Final mean hemoglobin and packed cell volume were 8.53 g/dl and 26.1%. Forty-three percent of the calves at d 0 were colostral deficient, assuming that total serum protein concentrations of <5.5 g/dl indicate colostral deficiency. No blood trait was consistently correlated with body weight gain when gain during the production period was divided into quartiles and the blood traits were averaged by gain quartile. Calves in the lowest serum total protein quartile (mean 4.58 g/dl) had more respiratory and total medical treatments than quartiles with higher total protein means. Dairy bull calves arriving at veal production units after transporting from the dairy farm to the auction market (or other collection facility) have several physical impairments. However, most of these physical impairments are improved early in the veal feeding period and are not generally related to subsequent growth rate or medical treatment. PMID- 10791803 TI - Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Europe and the United States. AB - Minimum inhibitory concentrations were determined for 811 strains of Staphylococcus aureus isolated from cases of bovine mastitis in 11 countries. The countries and number of isolates included Denmark (105), England (92), Finland (95), Germany (103), Iceland (22), Ireland (42), Norway (101), Sweden (123), Switzerland (69), United States (53), and Zimbabwe (6). The antimicrobial agents tested were penicillin, ampicillin, oxacillin, cephalothin, ceftiofur, amoxicillin + clavulanate, penicillin + novobiocin, enrofloxacin, premafloxacin, erythromycin, clindamycin, lincomycin, pirlimycin, neomycin, lincomycin + neomycin, and sulfamethazine. The MIC90 for these antimicrobial agents for all strains were 0.5, 1.0, 1.0, 0.5, 1.0, < or =0.06, 0.125, 0.125, < or =0.0078, 0.5, 1.0, 16.0, 1.0, 2.0, 0.5, and 4.0 microg/ml, respectively. Overall, only small variations between countries were seen in the MIC90 for the majority of compounds tested. Of the strains tested, 35.6% were positive for beta-lactamase production on initial testing, with an additional 21.3% positive after induction by penicillin. In conclusion, the overall level of resistance was generally low for all antimicrobial agents tested regardless of country. Given the differences in antimicrobial use in various countries, the widespread adoption of mastitis control programs to prevent infections limits the exposure of S. aureus infected animals to antimicrobial drugs. PMID- 10791804 TI - Invited review: effects of diet shifts on Escherichia coli in cattle. AB - Escherichia coli O157:H7 is a pathogenic bacterium that causes acute illness in humans, but mature cattle are not affected. E. coli O157:H7 can enter the human food supply from cattle via fecal contamination of beef carcasses at slaughter. Previous attempts to correlate the incidence of E. coli O157:H7 with specific diets or feeding management practices gave few statistically significant or consistent findings. However, recent work indicates that cattle diets may be changed to decrease fermentation acid accumulation in the colon. When fermentation acids accumulate in the colon and pH decreases, the numbers of acid resistant E. coli increase; acid-resistant E. coli are more likely to survive the gastric stomach of humans. When cattle were fed hay for a brief period (<7 d), acid-resistant E. coli numbers declined dramatically. Other workers have shown that brief periods of hay feeding can also decrease the number of cattle shedding E. coli O157:H7, and a similar trend was observed if cattle were taken off feed and exposed to simulated transport. These observations indicate that cattle feeding management practices may be manipulated to decrease the risk of foodborne illness from E. coli, but further work will be needed to confirm these effects. PMID- 10791805 TI - Hormones, mammary growth, and lactation: a 41-year perspective. AB - When I was a beginning graduate student 41 yr ago it had been established that estrogen caused mammary duct growth; a combination of estrogen and progesterone was required for lobule-alveolar development of the mammary glands; and prolactin and growth hormone were essential for mammary growth. In laboratory species exogenous prolactin, glucocorticoids, and estrogen would initiate secretion of milk provided the mammary glands had a well-developed lobule-alveolar system. It was not known with certainty that progesterone inhibited the process. For some species, prolactin and thyroxine had been shown to stimulate lactation, while glucocorticoids suppressed lactation. Definitive roles for growth hormone and insulin during lactation had not been established. Studies of hormonal control of mammary growth and function in cattle were few. In vitro methods to study hormonal regulation of the mammary glands were in their infancy. Quantitative measures of changes in mammary cell numbers and specific components of milk in response to hormones were rare. The concepts for quantification of hormone concentrations, hormone receptors, growth factors, and binding proteins in blood; hormonal regulation of nutrient partitioning; and hormonally induced mechanisms of action within mammary cells were waiting to be discovered. And eventually they were. However, lest we become too enamored with our current understanding of the hormones that control mammary growth and lactation, it remains a fact that the greatest physiological stimulus for milk yield is pregnancy, not some cocktail of exogenous hormones, growth factors, receptor agonists/antagonists, or gene therapies. Viva la mom! PMID- 10791806 TI - Photoperiodic effects on dairy cattle: a review. AB - Since the initial report in 1978 of galactopoietic effects of a photoperiod of 16 h of light:8 h of darkness, numerous studies have confirmed long-day stimulation of milk yield. The endocrine factor(s) responsible for increased milk yield, however, has eluded identification. Recent studies suggest that insulin-like growth factor-I (IGF-I) may mediate the galactopoietic response to long day photoperiod. Indeed, long days increase IGF-I in heifers and lactating cows; in the latter case, the response preceded an increase in milk yield. In heifers and cows, the increase in IGF-I is independent of changes in circulating growth hormone. Melatonin feeding to mimic a short-day photoperiod suppressed the increase of IGF-I in heifers induced by long days. However, melatonin feeding had no effect on milk yield in cows. Despite lack of resolution of the endocrine mechanism, dairy producers are interested in how photoperiod management can be integrated with current practices throughout the lactation cycle. There is strong evidence that milk yield responses to long days persist through an entire lactation. Also, long days can be combined with bovine somatotropin (bST) to produce additive increases in milk yield. During the dry period, long days increase the periparturient surge of prolactin. However, relative to long days, short-day treatment during the dry period produces the largest magnitude of response in milk yield during the subsequent lactation. The response to short days during the dry period may be due to a priming effect on the photoperiodic response system. In summary, IGF-I has emerged as a possible mediator of the increase of milk yield in response to long-day photoperiod. Photoperiod can be combined effectively with other management techniques such as bST. Consideration of photoperiod management during the dry period is essential to maximize responses during the subsequent lactation. PMID- 10791807 TI - Probiotic bacteria: selective enumeration and survival in dairy foods. AB - A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus casei. Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods. However, studies have shown low viability of probiotics in market preparations. In order to assess viability of probiotic bacteria, it is important to have a working method for selective enumeration of these probiotic bacteria. Viability of probiotic bacteria is important in order to provide health benefits. Viability of probiotic bacteria can be improved by appropriate selection of acid and bile resistant strains, use of oxygen impermeable containers, two-step fermentation, micro-encapsulation, stress adaptation, incorporation of micronutrients such as peptides and amino acids and by sonication of yogurt bacteria. This review will cover selective enumeration and survival of probiotic bacteria in dairy foods. PMID- 10791808 TI - Development of a PCR marker for rapid identification of the Bt-10 gene for common bunt resistance in wheat. AB - In western Canada, the Bt-10 resistance gene in wheat (Triticum aestivum) is effective against all the known races of common bunt caused by Tilletia tritici and T laevis. The genotypes of 199 F2 plants, originated from a cross between BW553 containing Bt-10 and the susceptible spring wheat cultivar 'Neepawa,' were established in greenhouse and field inoculation studies. A ratio of 1:2:1 resistant : heterozygous : susceptible was observed for bunt reaction, indicating that Bt-10 was expressed in a partially dominant fashion. A polymorphic DNA fragment, amplified using RAPD, and previously shown to be linked to Bt-10 was sequenced and SCAR (sequence characterized amplified region) primers devised. However, SCAR primers failed to amplify the polymorphic fragment. Restriction of PCR products with DraI revealed a polymorphic fragment of 490 bp resulting from a single base pair difference between lines possessing Bt-10 and those lacking the gene. As per the base pair difference, FSD and RSA primers were designed to generate a 275-bp polymorphic DNA fragment. Both 275- and 490-bp polymorphic fragments were present in all of the 22 cultivars known to carry Bt-10, and absent in all 16 cultivars lacking Bt-10. A 3:1 ratio was observed for presence: absence of the 275-bp marker in the F2 population. Using Southern analysis, the 490-bp fragment was effective in differentiating homozygous resistant plants from those heterozygous for Bt-10, based on its presence and the hybridization signal strength. A 1:2:1 resistant : heterozygous : susceptible ratio was also observed for the molecular marker and corresponded to 88% of the phenotypes deduced from the original F2 population. The molecular marker was estimated to be between 1.1 cM and 6.5 cM away from the Bt-10 resistance gene, based on the segregation analysis. Segregation analyses of Bt-10 and the 275-bp marker, evaluated in three different Canada Prairie Spring (CPS) wheat populations, demonstrated a segregation ratio of 3:1 for the molecular marker in two of the populations. These results demonstrated that the PCR marker system using the FSD and RSA primer pair permitted a rapid and reliable identification of individual lines carrying the Bt-10 gene for resistance to common bunt. PMID- 10791809 TI - Inheritance and RAPD tagging of multiple genes for resistance to net blotch in barley. AB - A doubled haploid barley (Hordeum vulgare L.) population that was created from a cross between cultivars 'Leger' and 'CI 9831' was characterized by RAPD (random amplified polymorphic DNA) markers for resistance to isolate WRS857 of Pyrenophora teres Drechs. f. sp. maculata Smedeg., the causal agent of the spot form of net blotch. Resistance, which initially appeared to be conferred by a single gene from the approximate 1:1 (resistant : susceptible) segregation ratio of the doubled-haploid (DH) progeny, was found to be associated with three different genomic regions by RAPD analysis. Of 500 RAPD random primers that were screened against the parents, 195 revealed polymorphic bands, seven showed an association to the resistance in bulks, and these seven markers were mapped to three unlinked genomic regions. Two of these regions, one of which was mapped to chromosome 2, have major resistance genes. The third region has some homology to the chromosome 2 region. This study demonstrates the simultaneous location of markers for more than one gene governing a trait by using RAPD and bulked segregant analysis (BSA). PMID- 10791810 TI - Relationship between pachytene synapsis, metaphase I associations, and transmission of 2B and 4B chromosomes in rye. AB - 2B rye plants selected for high (H) or low (L) B transmission rate were studied at pachytene and metaphase I of meiosis to determine the relationship between synapsis, bivalents at metaphase I, and B transmission rate. The results show that the 2 B chromosomes (Bs) form bivalents at pachytene in both the H and L lines, whereas the frequency of bivalents at metaphase I is much higher in the H than in the L line. This demonstrates that B transmission is mainly related to the proper association of Bs at metaphase I, as well as that synapsis of the 2 Bs in the L line is normal, but the bivalent is not consolidated by a chiasma in most cases. Crosses were made between 2B plants of the H and L lines in all combinations (H x H, H x L, L x H, and L x L) to obtain 4B plants. Similarly, bivalent formation at pachytene and metaphase I was studied. The results show that 4B plants of the H x H and L x L classes differ significantly at pachytene and metaphase I since the former forms more bivalents. The heterozygous 4 Bs of the H x L and L x H classes show intermediate values. The relation H x H > H x L > L x H > L x L was consistently found for the variables transmission rate, bivalents at pachytene, bivalents at metaphase I, and B mean chiasma frequency. A maternal effect was also found. Our data suggest that there are two separate mechanisms acting upon synapsis and chiasma formation in H and L B chromosomes: (i) there is variable efficiency of the control of synapsis at early stages of meiosis; and (ii) there is variable efficiency of the control of the number of chiasmata. PMID- 10791811 TI - Linkage group alignment of sorghum RFLP maps using a RIL mapping population. AB - Several molecular maps have been constructed in sorghum (Sorghum bicolor L. Moench) using a variety of probes from different grass species such as sorghum, maize, sugarcane, rice, oat, and barley. In order to enhance the utility of the existing mapping information by the sorghum research community, alignment and integration of all major molecular maps is necessary. To achieve this objective, a genetic map of 214 loci with a total map distance of 1200 cM was constructed using 98 F7 sorghum recombinant inbred lines (RILs) from a cross between two inbred lines, B35 and Tx7000. Few cDNA clones of sorghum and maize related to photosynthesis and drought stress were mapped on this map for the first time. Five major restriction fragment length polymorphism (RFLP) maps independently developed in this species were used for alignment purpose. The distributions of previously mapped markers were compared with their respective sorghum maps to align each of the linkage groups. In general, consistent linear order among markers was maintained in all the linkage maps. The successful alignment of these RFLP maps will now allow selection of a large number of markers for any region of the sorghum genome with many potential applications ranging from fine mapping and marker-assisted selection to map-based cloning for the improvement of sorghum and related species. PMID- 10791812 TI - Identification of a 5S rDNA spacer type specific Triticum urartu and wheats containing the T. urartu genome. AB - A PCR system was designed to amplify 5S spacer rDNA specifically from homeologous chromosome 1 in a variety of species representative of the Aegilops and Triticum genera. Two polymerase chain reaction (PCR) primer combinations were used, one of which appears to be apomorphic in nature and specific to chromosome 1A in Triticum urartu and tetraploid and hexaploid wheats containing the AA genome donated by T. urartu. The value of studying single repeat types to investigate the molecular evolution of 5S-rDNA arrays is considered. PMID- 10791813 TI - Extraordinary and extensive karyotypic variation: a 48-fold range in chromosome number in the gall-inducing scale insect Apiomorpha (Hemiptera: Coccoidea: Eriococcidae). AB - Chromosome number reflects strong constraints on karyotype evolution, unescaped by the majority of animal taxa. Although there is commonly chromosomal polymorphism among closely related taxa, very large differences in chromosome number are rare. This study reports one of the most extensive chromosomal ranges yet reported for an animal genus. Apiomorpha Rubsaamen (Hemiptera: Coccoidea: Eriococcidae), an endemic Australian gall-inducing scale insect genus, exhibits an extraordinary 48-fold variation in chromosome number with diploid numbers ranging from 4 to about 192. Diploid complements of all other eriococcids examined to date range only from 6 to 28. Closely related species of Apiomorpha usually have very different karyotypes, to the extent that the variation within some species-groups is as great as that across the entire genus. There is extensive chromosomal variation among populations within 17 of the morphologically defined species of Apiomorpha indicating the existence of cryptic species-complexes. The extent and pattern of karyotypic variation suggests rapid chromosomal evolution via fissions and (or) fusions. It is hypothesized that chromosomal rearrangements in Apiomorpha species may be associated with these insects' tracking the radiation of their speciose host genus, Eucalyptus. PMID- 10791814 TI - The genes encoding granule-bound starch synthases at the waxy loci of the A, B, and D progenitors of common wheat. AB - Three genes encoding granule-bound starch synthase (wx-TmA, wx-TsB, and wx-TtD) have been isolated from Triticum monococcum (AA), and Triticum speltoides (BB), by the polymerase chain reaction (PCR) approach, and from Triticum tauschii (DD), by screening a genomic DNA library. Multiple sequence alignment indicated that the wx-TmA, wx-TsB, and wx-TtD genes had the same extron and (or) intron structure as the previously reported waxy gene from barley. The lengths of the three wx-TmA, wx-TsB, and wx-TtD genes were 2834 bp, 2826 bp, and 2893 bp, respectively, each covering 31 bp in the untranslated leader and the entire coding region consisting of 11 exons and 10 introns. The three genes had identical lengths of exons, except exonl, and shared over 95% identity with each other within the exon regions. The majority of introns were significantly variable in length and sequence, differing mainly in length (1-57 bp) as a result of insertion and (or) deletion events. The deduced amino acid sequence from these three genes indicated that the mature WX-TMA, -TSB, and -TTD proteins contained the same number of amino acids, but differed in predicted molecular weight and isoelectric point (pI) due to amino acid substitutions (13-18). The predicted physical characteristics of the WX proteins matched the respective proteins in wheat very closely, but the match was not perfect. Furthermore the exon5 sequences of the wx-TmA, wx-TsB, and wx-TtD genes were different from a cDNA encoding a waxy gene of common wheat previously reported. The striking difference was that an insertion of 11 amino acids occurred in the cDNA sequence that could not be observed in the exons of the A, B, and D genes. It was noted, however, that the 3' end of intron4 of these genes could account for the additional 11 amino acids. The sequence information from the available waxy genes identified the intron4-exon5-intron5 region as being diagnostic for sequence variation in waxy. The sequence variation in the waxy genes provides the basis for primer design to distinguish the respective genes in common wheat, and its progenitors, using PCR. PMID- 10791815 TI - TAS49--a dispersed repetitive sequence isolated from subtelomeric regions of Nicotiana tomentosiformis chromosomes. AB - We have isolated and characterized a new repetitive sequence, TAS49, from terminal restriction fragments of Nicotiana tomentosiformis genomic DNA by means of a modified vectorette approach. The TAS49 was found directly attached to telomeres of N. tabacum and one of its ancestors, N. tomentosiformis, and also at inner chromosome locations. No association with telomeres was detected neither in N. otophora nor in the second tobacco ancestor, N. sylvestris. PCR and Southern hybridization reveal similarities in the arrangement of TAS49 on the chromosomes of 9 species of the genus Nicotiana, implying its occurrence as a subunit of a conserved complex DNA repeat. TAS49 belongs to the family of dispersed repetitive sequences without features of transposons. The copy number of TAS49 varies widely in the genomes of 8 species analyzed being lowest in N. sylvestris, with 3300 copies per diploid genome. In N. tomentosiformis, TAS49 forms about 0.56% of the diploid genome, corresponding to 17400 copies. TAS49 units are about 460 bp long and show about 90% of mutual homology, but no significant homology to DNA sequences deposited in GenBank and EMBL. Although genomic clones of TAS49 contain an open reading frame encoding a proline-rich protein similar to plant extensins, no mRNA transcript was detected. TAS49 is extensively methylated at CpG and CpNpG sites and its chromatin forms nucleosomes phased with a 170 +/- 8 bp periodicity. PMID- 10791816 TI - Varied expression of a Y-linked P[w+] insert due to imprinting in Drosophila melanogaster. AB - During gametogenesis, a gene can become imprinted affecting its expression in progeny. We have used the expression of a Y-linked P[w+]YAL transposable DNA element as a reporter system to investigate the effect of parental origination on the expression of the w+ insert. Expression of w+ was greater in male progeny when the Y chromosome, harboring the insert, was inherited from the parental male rather than from the parental female. Imprinting was not due to a genetic background influence in the males, since the only difference among the males was the parental origin of the Y chromosome. It was also observed that the genetic background can affect imprinting, since w+ expression was also higher in males when the Y was derived from C(1)DX attached-X parental females rather than from C(1)RM attached-X parental females. Though the heterochromatic imprinting mechanism is unknown, a mutated Heterochromatin Protein 1 (HP1) gene, which is associated with suppression of position-effect variegation, increases expression of the w+ locus in the P[w+]YAL insert, indicating that HP1 may play a role in Y chromosome packaging. PMID- 10791817 TI - Microsatellite DNA markers in Populus tremuloides. AB - Markers for eight new microsatellite DNA or simple sequence repeat (SSR) loci were developed and characterized in trembling aspen (Populus tremuloides) from a partial genomic library. Informativeness of these microsatellite DNA markers was examined by determining polymorphisms in 38 P. tremuloides individuals. Inheritance of selected markers was tested in progenies of controlled crosses. Six characterized SSR loci were of dinucleotide repeats (two perfect and four imperfect), and one each of trinucleotide and tetranucleotide repeats. The monomorphic SSR locus (PTR15) was of a compound imperfect dinucleotide repeat. The primers of one highly polymorphic SSR locus (PTR7) amplified two loci, and alleles could not be assigned to a specific locus. At the other six polymorphic loci, 25 alleles were detected in 38 P. tremuloides individuals; the number of alleles ranged from 2 to 7, with an average of 4.2 alleles per locus, and the observed heterozygosity ranged from 0.05 to 0.61, with an average of 0.36 per locus. The two perfect dinucleotide and one trinucleotide microsatellite DNA loci were the most informative. Microsatellite DNA variants of four SSR loci characterized previously followed a single-locus Mendelian inheritance pattern, whereas those of PTR7 from the present study showed a two-locus Mendelian inheritance pattern in controlled crosses. The microsatellite DNA markers developed and reported here could be used for assisting various genetic, breeding, biotechnology, genome mapping, conservation, and sustainable forest management programs in poplars. PMID- 10791818 TI - Chromosomal location of AFLP markers in common wheat utilizing nulli-tetrasomic stocks. AB - Amplified fragment length polymorphism (AFLP) markers with a total of 256 EcoRI + ANN - MseI + CNN primer combinations were investigated employing the common wheat cultivar Triticum aestivum 'Chinese Spring.' On average, 103 fragments per primer combination were amplified, ranging from a maximum of 226 fragments to a minimum of 18 fragments. The primer combinations E + AAA - M + CNN and E + ATT - M + CNN produced very few distinct fragments. By using 15 randomly chosen EcoRI + ANN - MseI + CNN primer combinations, 928 AFLP markers were allocated to wheat chromosomes, of which 131 were assigned to specific chromosome arms. These AFLP markers were locus-specific and randomly distributed on the different chromosomes. In addition, 6 and 41 AFLP markers were simultaneously absent in two nulli-tetrasomics (NTs) of both homoeologous and non-homoeologous groups, respectively, whereas additional fragments were detected in N1BT1A, N5AT5D, and N6BT6A lines. PMID- 10791819 TI - Cloning of two glutamate dehydrogenase cDNAs from Asparagus officinalis: sequence analysis and evolutionary implications. AB - Two different amplification products, termed c1 and c2, showing a high similarity to glutamate dehydrogenase sequences from plants, were obtained from Asparagus officinalis using two degenerated primers and RT-PCR (reverse transcriptase polymerase chain reaction). The genes corresponding to these cDNA clones were designated aspGDHA and aspGDHB. Screening of a cDNA library resulted in the isolation of cDNA clones for aspGDHB only. Analysis of the deduced amino acid (aa) sequence from the full-length cDNA suggests that the gene product contains all regions associated with metabolic function of NAD glutamate dehydrogenase (NAD-GDH). A first phylogenetic analysis including only GDHs from plants suggested that the two GDH genes of A. officinalis arose by an ancient duplication event, pre-dating the divergence of monocots and dicots. Codon usage analysis showed a bias towards A/T ending codons. This tendency is likely due to the biased nucleotide composition of the asparagus genome, rather than to the translational selection for specific codons. Using principal coordinate analysis, the evolutionary relatedness of plant GDHs with homologous sequences from a large spectrum of organisms was investigated. The results showed a closer affinity of plant GDHs to GDHs of thermophilic archaebacterial and eubacterial species, when compared to those of unicellular eukaryotic fungi. Sequence analysis at specific amino acid signatures, known to affect the thermal stability of GDH, and assays of enzyme activity at non-physiological temperatures, showed a greater adaptation to heat-stress conditions for the asparagus and tobacco enzymes compared with the Saccharomyces cerevisiae enzyme. PMID- 10791820 TI - RAPD analysis of seed purity in a commercial hybrid cabbage (Brassica oleracea var. capitata) cultivar. AB - The use of random amplified polymorphic DNA (RAPD) markers for evaluating seed purity in a commercial F1-hybrid cabbage (Brassica oleracea var. capitata) cultivar is demonstrated. Genomic DNA isolated from single ungerminated seed was found to be suitable for RAPD analysis. DNA from F1-hybrid and its parental lines was subjected to RAPD screening with 36 random decamer arbitrary primers. A total of 241 scorable products were observed with 54 (22%) being polymorphic. The RAPD data showed that the parental lines of this commercial cabbage cultivar were not very closely related. Two primers were chosen for purity testing of the F1-hybrid seeds. The sib (inbred seed; seed from self-pollination of parental lines) contamination results obtained by RAPD analysis were comparable to the commonly used grow-out trial and isozyme analysis, hence showing that RAPD analysis can be used for seed purity testing of commercial hybrid cabbage seeds. PMID- 10791821 TI - Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis. AB - Amplified fragment length polymorphism (AFLP) analysis was used to assess the biodiversity of one of the most important dipteran pests of cereals, the Asian rice gall midge (Orseolia oryzae Wood Mason). Larvae and pupae were collected at 15 locations in five Asian countries and preserved in 95% ethanol for storage, shipment, and DNA extraction using cetyltrimethylammonium bromide (CTAB). Although only approximately 1 microg of DNA was extracted from a single pupa or larva, the use of several AFLP primers in various combinations meant that this amount of DNA was sufficient to allow many DNA fingerprints to be made per individual. Fingerprints were sufficiently reproducible, especially during selective amplification, to allow the genetic diversity within a field population to be characterized. Extraction of DNA from a pool of 20 insects yielded AFLP fingerprints in which variation among individuals was sacrificed in favor of detecting differences among populations. For each location, pooled DNA was amplified with three primer pairs. A total of 261 distinct AFLP bands were identified for the 45 fingerprints. Cluster analysis, performed by the unweighted pair-group method (UPGMA), separated the populations into two distinct groups. Group I included two populations from Guangdong province of southern China and one each from Laos and Imphal in northeastern India, while group II was comprised of eleven populations from elsewhere in India (Assam, Orissa, Madhya Pradesh, Andhra Pradesh, and Kerala) and from Nepal and Sri Lanka. AFLP analysis provided insight into the origins of gall midge biotypes. In 1992, the prevailing biotype in Imphal changed from Indian biotype 3 to a new biotype 3M. Our data show that biotype 3M belongs to group I and did not arise by a recent mutation from biotype 3, which belongs to group II. By contrast, Indian biotypes 2 and 4 are likely to have diverged through recent mutation and selection, as are Chinese biotypes 1 and 4. The almost simultaneous emergence of new biotypes in Kerala and Sri Lanka during 1985-1988 was most probably coincidental, because these biotypes are not closely related. AFLP fingerprints were also able to detect sexual dimorphism in the DNA of adult gall midges and to distinguish gall midge from its major parasite Platygaster oryzae. PMID- 10791822 TI - A gene controlling sex in grapevines placed on a molecular marker-based genetic map. AB - Genetic maps of Vitis (2n = 38) have been constructed from an interspecific hybrid population of 58 seedlings of the cross 'Horizon' ('Seyval' x 'Schuyler') x Illinois 547-1 (V. cinerea B9 x V. rupestris B38). The maps were initially constructed based on 277 RAPD (random amplified polymorphic DNA) markers using a double-pseudotestcross strategy. Subsequently, 25 microsatellites, 4 CAPS (cleaved amplified polymorphic sequence), and 12 AFLP (amplified fragment length polymorphism) markers were added to the maps. Another 120 markers, mostly those segregating 3:1, were also assigned but not positioned on the linkage groups in the two maps. The 'Horizon' map consisted of 153 markers covering 1199 cM, with an average map distance of 7.6 cM between markers. The Illinois 547-1 map had 179 markers covering 1470 cM, with an average map distance of 8.1 cM. There were 20 linkage groups in each map, one more than the basic number of chromosomes in grapes. Ten linkage groups in each map were identified as homologous using 16 microsatellite and 2 CAPS markers polymorphic in both parents. A single locus controlling sex in grapes mapped close to a microsatellite marker. These maps provide enough coverage of the genome for QTL (quantitative trait loci) analysis and as a starting point for positional gene cloning in grapes. PMID- 10791823 TI - Organization, nucleotide sequence, and chromosomal mapping of a tandemly repeated unit containing the four core histone genes and a 5S rRNA gene in an isopod crustacean species. AB - A tandemly repeated unit of 6553 bp containing a copy of the four core histone genes H2B, H2A, H3, and H4, and also a 5S rRNA gene, was amplified by PCR from genomic DNA of the isopod crustacean Asellus aquaticus. The linkage between 5S rRNA genes and histone genes has been so far observed in only one other organism, the anostrac crustacean Artemia salina. The gene cluster was cloned and sequenced. The histone genes, in their 3' flanking region, have the interesting feature of possessing two different mRNA termination signals, the stem-loop structure and the AATAAA sequence. A part of the PCR product was used as a probe in FISH experiments to locate the gene cluster on an inter-individually variable number of chromosomes from 6 to 12 per diploid cell, always in a terminal position and never associated with the heterochromatic areas. Fluorescence in situ hybridization (FISH) was also performed on preparations of released chromatin and the reiteration level of the gene cluster was determined as approximately 200-300 copies per haploid genome. PMID- 10791824 TI - Cloning and characterization of an extremely conserved satellite DNA family from the root-knot nematode Meloidogyne arenaria. AB - A new satellite DNA family, named pMaE, has been cloned from the genome of the phytoparasitic nematode, Meloidogyne arenaria (Nematoda: Tylenchida). It is represented as tandemly repeated sequences with a monomeric unit of 172 bp. The monomers are present at approximately 15700 copies per haploid genome, and represent about 5.3% of the total genomic DNA. Twenty-seven independent monomers have been cloned and sequenced. The deduced consensus sequence is 70.9% A + T rich, with frequent stretches of A and (or) T. Several direct or inverted sub repeats are present in the sequence, which may allow the formation of a dyad structure, suggesting some potential role of this repetitive sequence in heterochromatin condensation. The monomers are very homogeneous in sequence, showing on average 1.8% divergence from their consensus sequence. Moreover, Southern blot experiments and sequence analysis of homologous monomers from the genome of geographically distinct M. arenaria populations have shown that this satellite DNA is uniformly distributed and highly conserved within the species. Therefore, it is hypothesized that this unusually low level of variability, either within the genome of a given population or between populations, could be achieved as the result of some highly effective homogenization mechanism acting upon the nematode genome. PMID- 10791825 TI - Sequence analysis of trinucleotide repeat microsatellites from an enrichment library of the equine genome. AB - Microsatellites are useful tools for the construction of a linkage map and parentage testing of equines, but only a limited number of equine microsatellites have been elucidated. Thus, we constructed the equine genomic library enriched for DNA fragments containing (CAG)n repeats. The enriched method includes hybridization-capture of repeat regions using biotin-conjugated oligonucleotides, nucleotide substrate-biased polymerase reaction with the oligonucleotides and subsequent PCR amplification, because these procedures are useful for the cloning of less abundant trinucleotide microsatellites. Microsatellites containing (CAG)n repeats were obtained at the ratio of one per 3-4 clones, indicating an enrichment value about 10(4)-fold, resulting in less time consumption and less cost for cloning. In this study, 66 different microsatellites, (CAG)n repeats, were identified. The number of complete simple CAG repeats in our clones ranged 4 33, with an average repeat length of 8.8 units. The microsatellites were useful as sequence-tagged site (STS) markers. In addition, some clones containing (CAG)n repeats showed homology to human (CAG)n-containing genes, which have been previously mapped. These results indicate that the clones might be a useful tool for chromosome comparison between equines and humans. PMID- 10791826 TI - Cloning and characterization of Vine-1, a LTR-retrotransposon-like element in Vitis vinifera L., and other Vitis species. AB - We report the organization of a grapevine chimeric gene Adhr-Vine-1, composed by an Adhr gene, into which a retroelement, Vine-1, was inserted. Sequence analysis revealed that Adhr is a member of the Adh multigene family, but does not correspond to any other grapevine Adh described to date. Vine-1, albeit defective, is the most complete LTR (long terminal repeat)-retrotransposon-like element described in Vitis vinifera L. It is 2392 bp long, with two almost identical LTRs (287 bp) in the same orientation, and flanked by direct repeats of a 5 bp host DNA. This element presents other features, characteristic of retroviruses and retrotransposons including inverted repeats, a primer binding site, and a polypurine tract. It has a single open reading frame (ORF) of 581 amino acids, potentially encoding for a gag protein and parts of the protease and integrase proteins. Vine-1 is most likely related to the copia-like type family, but with no significant similarity to any previously described plant retrotransposon or inserted element, nor to any eukaryotic element described to date. Vine-1 element has been found in Adhr at the same location in different V. vinifera cultivars, but not in some other analyzed Vitis species. These data suggest that Vine-1 insertion in Adhr is specific to V. vinifera, and has occurred after the Adh isogene separation, but prior to cultivar development. Sequences related to Vine-1 were revealed in multiple copies in the V. vinifera genome and, to a lesser extent, in other analyzed Vitis species. The polymorphism observed prompts us to question the role played by transposition in the evolution of the Vitis genus. PMID- 10791827 TI - Chromosomal location of a Triticum timopheevii--derived powdery mildew resistance gene transferred to common wheat. AB - A dominant powdery mildew resistance gene introduced from Triticum timopheevii in line 146-155-T of common wheat, Triticum aestivum, was located on chromosome 6B by monosomic analysis. Restriction fragment length polymorphism (RFLP) and microsatellite analyses detected the presence of a T. timopheevii segment, translocated to chromosome 6B, with breakpoints between the loci Xpsr8/Xpsr964 on 6BS and Xpsr154/Xpsr546 on 6BL. The novel powdery mildew resistance gene, which has been designated Pm27, was shown to cosegregate with the microsatellite locus Xpsp3131, which is located on the introgressed T. timopheevii segment. The molecular data confirm the location of Pm27 on the translocated 6B chromosome. PMID- 10791828 TI - Sequence conservation and expression of the sex-lethal homologue in the fly Megaselia scalaris. AB - Sex-lethal (Sxl) is Drosophila melanogaster's key regulating gene in the sex determining cascade. Its homologue in Megaselia scalaris, the chromosome 3 gene Megsxl, codes for a protein with an overall similarity of 77% with the corresponding D. melanogaster sequence. Expression in M. scalaris, however, is very unlike that in D. melanogaster. Megsxl transcripts with a long ORF occur in both sexes. Differential splicing is conserved but not sex-specific. There are several splice variants, among them one is common to gonads and somatic tissues of all developmental stages investigated, one is specific for ovaries and embryos, and a third one is not found in ovaries. In the ovary, Megsxl is heavily transcribed in nurse cells and transported into eggs. These results suggest a non sex-determining function during early embryogenesis; the presence of Megsxl RNA in testes and somatic tissues calls for other (or more) functions. PMID- 10791829 TI - Molecular cytogenetic analysis of supernumerary heterochromatic segments in Rumex acetosa. AB - The dioecious plant Rumex acetosa shows intraspecific karyotype variation, caused by supernumerary heterochromatic segments or DAPI (4',6-diamidino-2 phenylindole) bands at the ends of the short arms of three pairs of autosomes. A DNA sequence (RAE730) specific to the supernumerary heterochromatic segments was cloned and sequenced. RAE730 was about 730 bp and AT-rich (71% AT-content). Using fluorescence in situ hybridization (FISH), RAE730 was localized in the supernumerary DAPI-positive heterochromatic segments on several mitotic chromosomes and chromocenters in interphase nuclei, but not in the DAPI-bands of Y or B chromosomes. RAE730 was tandemly arranged in the genome, and the copy number varied between plants from 40000 to 304000 copies per 2C, corresponding to the relative amount of supernumerary heterochromatic segments per genome. These results indicate that the karyotype variation caused by the supernumerary heterochromatic segment was generated by amplification or reduction of the tandem repeats of RAE730. PMID- 10791830 TI - Genomic in situ hybridization (GISH) reveals high chromosome pairing affinity between Lolium perenne and Festuca mairei. AB - Intergeneric hybridizations have been made between species of Lolium and Festuca. It has been demonstrated, largely through conventional cytogenetic analysis, that the genomes of the two genera are related, however, much information is lacking on exactly how closely related the genomes are between the two species. We applied genomic in situ hybridization (GISH) techniques to the F1 hybrids of tetraploid Festuca mairei with a genomic constitution of M1M1M2M2 and diploid Lolium perenne with a genomic constitution of LL. It was shown in the triploid hybrids (LM1M2) that the chromosomes of M1 and M2 from F. mairei could pair with each other, and it was further discovered that L chromosomes of L. perenne paired with M1 and M2 chromosomes. Our results showed that meiocytes of Lolium-Festuca are amenable to GISH analysis, and provided direct evidence for the hypothesis that the chromosomes of Lolium and Festuca may be genetically equivalent and that reciprocal mixing of the genomes may be possible. PMID- 10791831 TI - Identification and chromosomal localization of a transcriptionally active retrotransposon of Ty3-gypsy type in rice. AB - A DNA fragment representing a transcriptionally active retrotransposon of Ty3 gypsy type was isolated and characterized from rice (Oryza sativa L.). The fragment (named RIRE9) includes the coding sequences for the C-terminal part of the RNase H domain and the N-terminal part of the integrase domain in the polyprotein region. Northern blot hybridization indicated that this element was expressed in rice leaves and stems, suggesting that it is potentially active to transpose under normal growth conditions. Using dot-blot hybridization, the copy number of RIRE9 was estimated to be about 1600 copies per haploid rice genome. Five homologous copies of RIRE9 were assigned to five distinct positions of four chromosomes by restriction fragment length polymorphism (RFLP) mapping approach using an indica-japonica rice doubled-haploid (DH) population and its molecular linkage map. PMID- 10791832 TI - Polyploidy in a natural population of mussel, Mytilus trossulus. AB - We have analyzed natural polyploidy in a population of Mytilus trossulus from Vancouver Island (British Columbia, Canada) by means of cytogenetic techniques. Results obtained are the first reporting on this type of numerical chromosome aberrations in mussels. PMID- 10791833 TI - Isolation of large terminal sequences of BAC inserts based on double-restriction enzyme digestion followed by anchored PCR. AB - Isolation of the terminal portions of genomic DNA cloned in bacterial artificial chromosomes (BACs) is an important step in map-based cloning, and several methods have been developed. Here, we present a new method based on double-restriction enzyme digestion followed by anchored PCR. BAC DNA was digested with two enzymes: NotI and one of four enzymes (EcoRV, HpaI, StuI, or XmnI) that produce blunt termini. After dephosphorylation, these digestions were ligated to NotI- and EcoRV-digested pMSK, a new cloning vector developed in this work that is derived from pBluescript SK(+). PCR products representing the left- and right-terminal sequences of BAC inserts were obtained using a primer complementary to pMSK and a primer complementary to sequences in either the left arm or the right arm of the BAC vector pBeloBAC11. We have tested this method with 15 different BAC clones, and PCR products representing both the left- and right-terminal sequences have been obtained from all 15 BAC clones. This method is simple, fast, reproducible, and uses the same set of primers for any restriction enzyme used. With some modifications, it can also be used for isolating the terminal portions of genomic DNA cloned in yeast artificial chromosomes and P1-derived artificial chromosomes. PMID- 10791834 TI - The parahippocampal region: basic science and clinical implications. PMID- 10791835 TI - Hemispheric differences in hippocampal volume predict verbal and spatial memory performance in patients with Alzheimer's disease. AB - Atrophy of the hippocampal formation, a region important for the acquisition of new declarative knowledge, has been well-documented in Alzheimer's disease (AD), although the relation of such atrophy to the extent of memory dysfunction in these patients has been less clear. In the present study, 18 patients with a clinical diagnosis of probable AD were studied with a high-resolution, quantitative magnetic resonance imaging (MRI) protocol, as well as the verbal and spatial versions of the Buschke controlled learning task. The volumes of the hippocampal formation and, as a control for generalized atrophy, parahippocampal gyrus and temporal neocortex were computed from gapless coronal slices taken perpendicular to the long axis of the hippocampus. To correct for individual differences in brain size, volumes of regions of interest were divided by total intracranial volume. Separate stepwise regression analyses (with age, right and left hippocampal, parahippocampal gyrus, and temporal lobe volumes as the independent variables) showed that left hippocampal volume was the best predictor of free recall and delayed free recall of verbal information (P = 0.0042 and P < 0.0001, respectively). Recall and delayed recall of the spatial location of verbal items were best predicted by right hippocampal volume (P = 0.0054 and P = 0.0118, respectively). Memory scores did not correlate either with parahippocampal gyrus or temporal lobe volume. Furthermore, the relation between hippocampal volume and memory function observed in cases with AD did not hold for healthy aged control subjects. PMID- 10791836 TI - Long-term potentiation in the reciprocal corticohippocampal and corticocortical pathways in the chronically implanted, freely moving rat. AB - The hippocampus and adjacent cortical structures, including the entorhinal, perirhinal, and parahippocampal cortices, appear to serve as an integrated memory system. This extended hippocampal system is believed to influence memory and consolidation through an extensive set of reciprocal connections with widespread areas of the neocortex. Long-term potentiation (LTP) has been well-examined in the intrinsic connections of the hippocampus and neocortex. However, LTP in the pathways and structures thought to convey information between the hippocampus and neocortex has received little attention. If these pathways and structures are involved in information storage, and if LTP reflects a general synaptic encoding mechanism, then these systems are also likely to support LTP. In this paper we discuss a series of experiments aimed at investigating LTP in the efferents between the hippocampus and neocortex in chronically implanted animals. In the first experiment, the efferents of the perirhinal cortex were stimulated. LTP in the dentate gyrus (DG) reached asymptote more slowly than is typically seen following perforant path stimulation, whereas the frontal area (M1) reached asymptote more quickly than reported following corticocortical stimulation. The DG and M1 LTP was long-lasting, but entorhinal cortex LTP had decayed to baseline levels after a week. In the second experiment, the hippocampal efferents were stimulated. The perirhinal, entorhinal, and frontal cortex showed a similar slow potentiation, with only the perirhinal cortex levels returning to baseline after a week. In the third experiment, the projections from M1 were tested. The perirhinal cortex and hippocampus showed a long-lasting LTP. Although LTP was found in all pathways examined, there were differences in the induction and decay rate, and these properties may correspond to differences in learning rate and longevity of information storage. PMID- 10791837 TI - Rats with hippocampal damage are impaired on place learning in the water task when overtrained under constrained conditions. AB - To date, numerous investigations have been conducted on the mammalian hippocampus to determine its precise function. This research has implicated a fundamental role for the hippocampus in the formation of a spatial map that an animal can use to appropriately guide behavior in complex relational tasks. Despite substantial evidence to support this view, there have been challenges to this theory of hippocampal function. One alternative view suggests that the hippocampus is involved with the integration and updating of voluntary movement. Therefore, any impairments expressed by rats with hippocampal damage are not due to the inability to form or use a spatial map, but rather arise because they are unable to accurately control and monitor on-line movement. Accordingly, investigators, supporting the latter, claim that animals with hippocampal lesions are able to solve a spatial version of the water task if they are given explicit training on how to get to the hidden platform. In the present study we trained rodents with or without hippocampal damage on a cue/place water task for 40 days. In using behaviorally constraining procedures and by overtraining these animals, we provided them with knowledge of how to get to the hidden platform, and ensured enough time to learn the task. Our findings revealed that although rats with hippocampal lesions showed some place responses, they were significantly impaired on all measures of place learning compared to sham animals under these intensive procedures. Overall, the results of the present study do not support the idea that the hippocampus is not specifically involved in acquisition of place information in the water task. PMID- 10791838 TI - Differential expression of cytoskeletal proteins in the dendrites of parvalbumin positive interneurons versus granule cells in the adult rat dentate gyrus. AB - Parvalbumin-positive interneurons and granule cells of the dentate gyrus exhibit characteristic differences in morphological, cytochemical, physiological, and pathophysiological properties. Several of these defining features, including dendritic morphology, spine density, and sensitivity to insults, are likely to be influenced by the neuronal cytoskeleton. The data in this paper demonstrate striking differences in the expression levels of all three neurofilament triplet proteins, as well as alpha-internexin and beta-tubulin III, between the parvalbumin-positive interneurons and dentate granule cells. Therefore, the molecular composition of intermediate filaments and microtubules in the dendritic domain of parvalbumin-positive dentate interneurons is distinct from the cytoskeleton of neighboring granule cells, indicating the existence of highly cell type-specific cytoskeletal architecture within the dentate gyrus. PMID- 10791839 TI - Cell damage and neurogenesis in the dentate granule cell layer of adult rats after pilocarpine- or kainate-induced status epilepticus. AB - Dentate granule cells are generally considered to be relatively resistant to excitotoxicity and have been associated with robust synaptogenesis after neuronal damage. Synaptic reorganization of dentate granule cell axons, the mossy fibers, has been suggested to be relevant for hyperexcitability in human temporal lobe epilepsy and animal models. A recent hypothesis suggested that mossy-fiber sprouting is dependent on newly formed dentate granule cells. However, we recently demonstrated that cycloheximide (CHX) can block the mossy-fiber sprouting that would otherwise be induced by different epileptogenic agents and does not interfere with epileptogenesis in those models. Here, we investigated cell damage and neurogenesis in the dentate gyrus of pilocarpine- or kainate treated animals with or without coadministration of CHX. Dentate granule cells were highly vulnerable to pilocarpine induced-status epilepticus (SE), but were hardly damaged by kainate-induced SE. CHX pretreatment markedly reduced the number of injured neurons after pilocarpine-induced SE. Induction of SE dramatically increased the mitotic rate of KA- and KA + CHX-treated animals. Induction of SE in animals injected with pilocarpine alone led to 2-7-fold increases in the mitotic rate of dentate granule cells as compared to 5- and 30 fold increases for pilocarpine + CHX animals. We suggest that such increased mitotic rates might be associated with a protection of a vulnerable precursor cell population that would otherwise degenerate after pilocarpine-induced SE. We further suggest that mossy-fiber sprouting and neurogenesis of granule cells are not necessarily linked to one another. PMID- 10791840 TI - Awareness predicts the magnitude of single-cue trace eyeblink conditioning. AB - Studies of differential eyeblink conditioning (CS+ and CS-) have demonstrated that successful conditioning requires awareness of the stimulus contingencies and that delay conditioning does not. Two experiments were carried out to determine whether awareness is also important for single-cue trace eyeblink conditioning. In experiment 1, participants who performed a secondary, attention-demanding task emitted significantly fewer conditioned eyeblink responses than participants who watched a silent movie during the conditioning session. In experiment 2, participants who became aware of the stimulus contingencies early in the conditioning session emitted significantly more conditioned responses during the remainder of the session than participants who became aware later in the session or who never became aware. These results indicate that awareness is important for single-cue trace eyeblink conditioning, just as it is for differential trace conditioning. The relationship between awareness and trace eyeblink conditioning is discussed in the light of these and other recent findings. PMID- 10791841 TI - Cholinergic induction of oscillations in the hippocampal slice in the slow (0.5-2 Hz), theta (5-12 Hz), and gamma (35-70 Hz) bands. AB - Carbachol, a muscarinic receptor agonist, produced three distinct spontaneous oscillations in the CA3 region of rat hippocampal slices. Carbachol concentrations in the 4-13 microM range produced regular synchronized CA3 discharges at 0.5-2 Hz (carbachol-delta). Higher concentrations (13-60 microM) produced short episodes of 5-10 Hz (carbachol-theta) oscillations separated by nonsynchronous activity. Concentrations of carbachol ranging from 8-25 microM also produced irregular episodes of high-frequency discharges (carbachol-gamma, 35-70 Hz), in isolation or mixed with carbachol-theta and carbachol-delta. At carbachol concentrations sufficient to induce carbachol-theta, low concentrations of APV reversibly transformed carbachol-theta into carbachol-delta. Higher concentrations of D,L-2-amino-5-phosphonopentanoic acid (APV) reversibly and completely blocked carbachol-theta. A systematic study of the effects of carbachol shows that the frequency of spontaneous oscillations depended nonlinearly on the level of muscarinic activation. Field and intracellular recordings from CA1 and CA3 pyramidal cells and interneurons during carbachol induced rhythms revealed that the hippocampal circuitry preserved in the slice was capable of spontaneous activity over the range of frequencies observed in vivo and suggests that the presence of these rhythms could be under neuromodulatory control. PMID- 10791843 TI - Why jump on estrogen? PMID- 10791842 TI - Voltage-clamp analysis of the potentiation of the slow Ca2+-activated K+ current in hippocampal pyramidal neurons. AB - Exploring the principles that govern activity-dependent changes in excitability is an essential step to understand the function of the nervous system, because they act as a general postsynaptic control mechanism that modulates the flow of synaptic signals. We show an activity-dependent potentiation of the slow Ca2+ activated K+ current (sl(AHP)) which induces sustained decreases in the excitability in CA1 pyramidal neurons. We analyzed the sl(AHP) using the slice technique and voltage-clamp recordings with sharp or patch-electrodes. Using sharp electrodes-repeated activation with depolarizing pulses evoked a prolonged (8-min) potentiation of the amplitude (171%) and duration (208%) of the sl(AHP). Using patch electrodes, early after entering the whole-cell configuration (<20 min), responses were as those reported above. However, although the sl(AHP) remained unchanged, its potentiation was markedly reduced in later recordings, suggesting that the underlying mechanisms were rapidly eliminated by intracellular dialysis. Inhibition of L-type Ca2+ current by nifedipine (20 microM) markedly reduced the sl(AHP) (79%) and its potentiation (55%). Ryanodine (20 microM) that blocks the release of intracellular Ca2+ also reduced sl(AHP) (29%) and its potentiation (25%). The potentiation of the sl(AHP) induced a marked and prolonged (>50%; approximately equals 8 min) decrease in excitability. The results suggest that sl(AHP) is potentiated as a result of an increased intracellular Ca2+ concentration ([Ca2+]i) following activation of voltage-gated L-type Ca2+ channels, aided by the subsequent release of Ca2+ from intracellular stores. Another possibility is that repeated activation increases the Ca2+ binding capacity of the channels mediating the sl(AHP). This potentiation of the sl(AHP) could be relevant in hippocampal physiology, because the changes in excitability it causes may regulate the induction threshold of the long-term potentiation of synaptic efficacy. Moreover, the potentiation would act as a protective mechanism by reducing excitability and preventing the accumulation of intracellular Ca2+ to toxic levels when intense synaptic activation occurs. PMID- 10791844 TI - Lung cancer surveillance: new technologies and novel strategies. PMID- 10791845 TI - Ambulatory breast cancer surgery. PMID- 10791847 TI - Ambulatory surgery for breast cancer patients. AB - BACKGROUND: Less than two decades ago, early discharge of mastectomy patients was found to be possible while the drains were still in place, without noticeable consequences for patients. Most reported studies focused on surgical complication rates and found no significant evidence of it. The objective of the present study was to compare inpatient to same-day discharge surgery for breast cancer, on unselected patients. METHODS: All interviewed patients (n = 90) had routine level I and II axillary lymph node dissection under general anesthesia, combined with breast surgery for most of them. The outpatient group comprised 55 patients and the inpatient group 35. Psychological distress was assessed, as well as pain, anxiety, quality of life, emotional adjustment, recovery, social relations, stressful life events, and so on. RESULTS: The sociodemographic characteristics of both surgery groups was quite similar, except that time from surgery to interview was about 1 year longer for inpatients. Outpatients and hospitalized patients report similar levels of pain, fear, anxiety, health assessment, and quality of life. Ambulatory patients manifest a significantly better emotional adjustment and fewer psychological distress symptoms. Inpatients reported that it took an average of 27 days to feel that they had recovered from surgery, about 10 days longer than outpatients. Inpatient return to usual activities was also about 11 days later. CONCLUSIONS: Same-day discharge patients are not at a disadvantage compared to hospitalized patients; i.e., they report faster recovery and better psychological adjustment. Outpatient surgery may thus foster patient emotional well-being better than routine hospitalization. PMID- 10791846 TI - Fluorescence bronchoscopic surveillance after curative surgical resection for non small-cell lung cancer. AB - BACKGROUND: Second lung primaries occur at a rate of up to 3% per patient-year after curative resection for non-small-cell lung carcinoma. Postresection patients are often poor candidates for further curative surgery because of their diminished pulmonary reserve. The aim of this study was to evaluate the role of fluorescence bronchoscopy by using the Xillix LIFE-Lung Fluorescence Endoscopy System to identify second lung primaries in patients who have had a previous curative resection of a non-small-cell lung cancer. METHODS: Patients who had no evidence of disease status after resection of a non-small-cell lung cancer were identified from a prospectively collected data base and entered onto a fluorescence bronchoscopy surveillance protocol. All suspicious areas, as well as several areas of apparently normal mucosa, were sampled for biopsy. A single pathologist reviewed all biopsy specimens, with 10% of biopsies re-reviewed, for quality control, by a second pulmonary pathologist. RESULTS: A total of 31 surveillance fluorescence bronchoscopies were performed on 25 patients after conventional bronchoscopy. Four intraepithelial neoplasias or invasive carcinomas were identified in 3 (12%) of 25 patients screened. The addition of the LIFE examination to conventional bronchoscopy increased the sensitivity of screening from 25.0% to 75.0%, which yielded a relative sensitivity of 300% with a negative predictive value of .97. CONCLUSIONS: Use of postresection surveillance with fluorescence bronchoscopy identified intraepithelial or invasive lesions in 12% of non-small-cell lung cancer patients, and the system was three times more sensitive than conventional bronchoscopy to identify these early mucosal lesions. Fluorescence bronchoscopic surveillance of this high-risk, postresection population will help better define the true rate of occurrence and the natural history of second primaries and may assist in monitoring their response to newer, noninvasive treatment methods, such as photodynamic therapy or chemopreventive agents, in future trials. PMID- 10791848 TI - Should internal mammary nodes be sampled in the sentinel lymph node era? AB - BACKGROUND: Controversy exists regarding internal mammary lymph nodes (IMNs) in the staging and treatment of breast cancer. Sentinel lymph node identification with radiocolloid can map drainage to IMNs and directed biopsy can be performed with minimal morbidity. Furthermore, recent studies suggest that IMN drainage of breast tumors may be underestimated. To gain further insight into the prognostic value of IMNs, we reviewed the outcome of patients in whom the IMN status was routinely assessed. METHODS: A retrospective review of 286 patients with breast cancer who underwent IMN dissection between 1956 and 1987 was conducted. RESULTS: Median follow-up is 186 months, age was 52 years (range, 21-85 years), tumor size was 2.5 cm, and number of IMNs removed was 5 (range, 1-22); 44% received chemotherapy, 16% endocrine therapy, and 5% radiotherapy. Presence of IMN metastases correlated with primary tumor size (P < .0001) and number of positive axillary nodes (P < .0001) but did not correlate with primary tumor location or age. Overall, the 20-year disease-free survival is significantly worse for the 25% of patients with IMN metastases (P < .0001). In patients with positive axillary nodes and tumors smaller than 2 cm, there was a significantly worse survival (P < .0001) in the patients with IMN metastases. This difference in survival was not seen in women with tumors larger than 2 cm. CONCLUSIONS: Patients with IMN metastases, regardless of axillary node status, have a highly significant decrease in 20-year disease-free survival. Treatment strategies based on knowledge of sentinel IMN status may lead to improvement in survival, especially for patients with small tumors. At present, sentinel IMN biopsies should be performed in a clinical trial setting. PMID- 10791849 TI - Relation between thymidylate synthase expression and survival in colon carcinoma, and determination of appropriate application of 5-fluorouracil by immunohistochemical method. AB - BACKGROUND: Thymidylate synthase (TS) is regarded as a parameter of 5 fluorouracil (5-FU) chemosensitivity for colorectal carcinoma. Recent researchers indicate that the chemosensitivity of 5-FU for colorectal carcinoma with low expression of TS is better than tumors with high expression of TS. But the relation between TS expression and overall survival of curatively resected colorectal cancer patients has been less studied. METHODS: Specimens of curatively resected colon carcinoma from 148 patients were included in this study. TS expression in the tumor was assessed by immunohistochemical staining technique, and the patients were categorized into TS-(+) and TS-(-) groups. First, the relation between TS expression and survival of patients was examined. Next, for each group, we compared survival between the chemotherapy-(+) and the chemotherapy-(-) subgroup. RESULTS: Overall survival was significantly better in the TS-(-) group (n = 107) than in the TS-(+) group (n = 41) (P = .0003). In the TS-(-) group, there was little difference between the chemotherapy-(+) and the chemotherapy-(-) subgroup. In the TS-(+) group, the survival of the chemotherapy (+) subgroup was significantly better than the chemotherapy-(-) subgroup (P = .0439). CONCLUSIONS: TS, itself, may be a prognostic factor for colon carcinoma; and 5-FU adjuvant chemotherapy may be appropriate for colon carcinoma with high expression of TS. PMID- 10791850 TI - Pleural extension of mucinous tumor in patients with pseudomyxoma peritonei syndrome. AB - BACKGROUND: Pseudomyxoma peritonei syndrome is a rare disease arising from perforation of an adenoma of the appendix. The syndrome is characterized by progressive accumulation of mucinous fluid and tumor within the abdomen and pelvis. Although this tumor is only superficially invasive and does not metastasize, it is a fatal disease. Extra-abdominal spread of pseudomyxoma peritonei is a rare occurrence, with few reports in the medical literature. This review focuses on pleural extension of mucinous tumor in patients with pseudomyxoma peritonei syndrome. METHODS: From December 1983 to April 1999, all patients who underwent cytoreductive surgery for pseudomyxoma peritonei syndrome were assessed for pleural involvement at the time of the presentation or follow up. Clinical information on these patients, including chest computed tomographic scan, was retrospectively reviewed. The mechanisms of extension of mucinous tumor from peritoneal cavity to pleural surface and the results of treatment were of special interest. RESULTS: Twenty-three of 426 patients (5.4%) showed pleural extension of pseudomyxoma peritonei syndrome. In four patients (17%), extension into the chest occurred before cytoreductive surgery. In 18 patients, the pleural space was entered during a subdiaphragmatic peritonectomy; and, in 12 patients, extension of disease from peritoneal to pleural space occurred. In six patients (26%), surgical interventions were required to excise tumor that had invaded the hemidiaphragm; and, in the six other patients (26%), there was a minor penetration during subphrenic peritonectomy, which was closed immediately. Finally, in seven patients (30%), the mechanism of spread was unknown. Twelve patients were treated for pleural thoracotomy. Eight patients had an attempt to completely eradicate pleural mucinous tumor, and five patients are currently disease free in the chest (22%); four of these five had intrapleural cytoreduction plus intrapleural chemotherapy. The median survival for all 23 patients is 55 months. CONCLUSION: Pleural spread of pseudomyxoma peritonei syndrome may be a direct result of cytoreductive surgery and the subphrenic peritonectomy procedure. In some patients, dissecting mucinous tumor may infiltrate through the diaphragm and result in pleural extension. Pleural extension of pseudomyxoma peritonei syndrome carries a poor prognosis. Intrapleural chemotherapy combined with cytoreductive surgery may be of considerable value in treatment and prevention of disease dissemination; it should be considered when pleural extension of mucinous tumor is feared or confirmed at the time of cytoreductive surgery. PMID- 10791851 TI - Biological evaluation of undifferentiated carcinoma of the esophagus. AB - BACKGROUND: Patients with undifferentiated carcinoma of the esophagus (UEC) are rare and have a poor prognosis compared with those with differentiated squamous cell carcinomas (DECs). We compared clinicopathological and biological features of UEC and DEC, with emphasis on markers for epithelial cell origin, proliferation, and cell-cell adhesion. METHODS: Seven patients with UEC were compared with 21 with DEC. Immunohistochemical studies were performed by using monoclonal antibodies to cytokeratin, epithelial membrane antigen, p53, p21WAF1/CIP1, Ki-67, E-cadherin, desmoglein-1, and thrombomodulin. RESULTS: Patients with UEC had a poorer prognosis because of hematogenous metastasis at the time of presentation (mean survival, 6.5 +/- 6.2 vs. 35.5 +/- 28.9 months; P < .05). Immunohistochemical findings for cytokeratin and epithelial membrane antigen suggest that some UECs had epithelial origins. The following immunohistochemical profile of UEC was consistent with its highly malignant properties: (1) reduced or negative expression of cell-cell adhesion molecules such as E-cadherin, desmoglein-1, and thrombomodulin, (2) high positive rate for p53 and Ki-67, and (3) negative expression of p21WAF1/CIP1. CONCLUSIONS: The immunohistochemical findings for UEC showed its high cell-proliferative activity and a high potential for metastasis. Clinical features of UEC were supported by the results of immunohistochemical findings. PMID- 10791852 TI - Survival benefit of D2 lympadenectomy in patients with gastric adenocarcinoma. AB - BACKGROUND: A definite resolution to the controversy on the optimal extension of lymph node dissection (LND) in gastric cancer has not been achieved. Surgical morbidity and survival of D1 and D2 LND are compared by multivariate analysis. METHODS: A retrospective cohort study of 219 patients with gastric cancer and curative resection performed according to Japanese rules. D1 dissection was performed in 106 cases and D2 in 113. The logistic regression model was used to define risk factors for surgical morbidity and the Cox model to determine prognostic factors. RESULTS: Surgical morbidity occurs in 16.9% and 19.5% in D1 and D2 LND, respectively (P = .7). The morbidity determinants were operation blood loss, splenectomy, pancreaticosplenectomy, antrum location, low serum albumin, total gastrectomy, and metastatic nodal ratio (P < .0001), but not D2 LND. Five-year survival was 35.1% for D1 and 64% for D2 LND (P < .039). The prognostic factors were T stage, N stage, serum albumin level, total gastrectomy, D2 LND, and comorbidity (P < .0001). CONCLUSIONS: The increment of surgical morbidity and mortality rates attributed to D2 LND is largely caused by the effect of splenectomy and pancreaticosplenectomy. A significant survival benefit because of D2 LND was found. The results support the value of extended LND in the surgical treatment of gastric cancer. PMID- 10791853 TI - Epithelioid sarcoma: the clinicopathological complexities of this rare soft tissue sarcoma. AB - BACKGROUND: Epithelioid sarcoma is a rare high grade soft tissue sarcoma with a known propensity for locoregional recurrence. The literature is limited on other characteristics such as frequency of multifocal disease at presentation, the relationship of presenting size of the primary lesion to prognosis, and the ability of current staging systems to predict prognosis. METHODS: Review of the Royal Marsden National Health Service Trust (RMH) experience of 37 cases over 21 years. RESULTS: The mean age was 29 years, with male predominance (2.7:1), and distal limb locations were most common (56%). Five patients presented with multifocal local disease. Median follow-up was 88 months in the 19 patients still alive. The 5- and 10-year actuarial overall survival was 70% and 42%, respectively. Tumors deep to the investing fascia had a worse prognosis than superficial tumors. Regional metastasis events were also associated with significantly worse overall survival. Local recurrence, size of 5 cm or larger, and regional metastasis events were predictive of worse distant metastasis-free survival. Tumor size (<5 cm vs. > or =5 cm), local recurrence events, sex, and site were not significant predictors of survival. The American Joint Committee on Cancer/International Union Against Cancer staging systems and the recently proposed RMH staging system of the Royal Marsden National Health Service Trust provided poor differentiation of prognosis in epithelioid sarcoma. The 5-year actuarial local recurrence rate was 35%. The 5-year actuarial regional nodal metastasis rate was 23%. The actuarial 5-year distant metastasis rate was 40%, with pleuropulmonary metastases the most common site of metastatic disease, and 35% of pleuropulmonary metastases presented with pleural effusion. Median post distant metastasis survival was 8 months. CONCLUSIONS: Epithelioid sarcoma has unusual clinical behavior compared with other high grade soft tissue sarcoma. It has a propensity for multifocal disease at presentation, local recurrence, regional metastasis, and particularly poor prognosis after regional or distant metastatic disease. Size and stage according to the American Joint Committee on Cancer/International Union Against Cancer are unreliable predictors of prognosis. PMID- 10791854 TI - Regional nodal basin control is not compromised by previous sentinel lymph node biopsy in patients with melanoma. AB - BACKGROUND: Regional nodal basin control is an important goal of lymphadenectomy in the management of melanoma patients with nodal disease. The purpose of this study was to determine if previous sentinel lymph node (SLN) biopsy compromises the ultimate regional nodal control achieved by subsequent therapeutic lymph node dissection in melanoma patients with microscopic lymph node metastases. METHODS: A surgical melanoma database and hospital records were reviewed for 602 patients with primary cutaneous melanoma who underwent successful lymphatic mapping and SLN biopsy between 1991 and 1997. RESULTS: A total of 105 (17%) of 602 patients had histologically positive SLNs and were offered therapeutic lymphadenectomy; 101 (96%) underwent this procedure. Thirty-six patients (36%) developed recurrent melanoma at one or more sites. The median follow-up period was 30 months. Recurrence in the previously dissected nodal basin was observed in 10 patients (10%); none had recurrence at only that site. Nodal basin disease appeared after local/in-transit (n = 6) or distant (n = 1) failure in seven patients and, as a component of the first site of failure, simultaneously with local/in-transit (n = 2) or distant (n = 1) recurrence in three patients. CONCLUSIONS: Nodal basin failure after lymphadenectomy in patients who underwent previous biopsy of a histologically positive SLN is primarily a function of aggressive locoregional disease rather than of contamination from previous surgery. Because regional nodal control was comparable with that in other series, we conclude that SLN biopsy with selective lymphadenectomy does not compromise regional nodal basin control. PMID- 10791855 TI - Does endogenous immune response determine the outcome of surgical therapy for metastatic melanoma? AB - BACKGROUND: Although the presence of tumor cells in the blood of patients with metastatic melanoma suggests widely disseminated disease, many of these patients enjoy prolonged survival or cure after surgical resection. Our previous study of adjuvant vaccine therapy after complete resection of metastatic melanoma revealed a strong correlation between postoperative survival and elevated antibody titers to a 90-kDa tumor-associated antigen (TA90) expressed by melanoma cells of the vaccine. We hypothesized a similar correlation between postoperative survival and endogenous anti-TA90 antibody titers induced by the patient's melanoma in the absence of postoperative adjuvant immunotherapy. METHODS: From 1970 to 1996, 64 patients underwent complete resection of distant melanoma metastases and did not receive postoperative adjuvant immunotherapy. Serum collected within 4 months after surgery was tested in a coded and blinded fashion for anti-TA90 IgG and IgM by enzyme-linked immunosorbent assay, and for total IgG and IgM (controls) by radial immunodiffusion. RESULTS: Median follow-up for the study population was 19 months (range, 3-147 months). There was no significant correlation between anti TA90 IgG titer and total IgG level (P = .4785), or between anti-TA90 IgM and total IgM (P = .0989). Univariate analysis showed that postoperative anti-TA90 IgM titer as a continuous variable was significantly associated with overall survival (OS); i.e., the higher the anti-TA90 IgM titer, the longer the OS. Using an established cutoff titer of 800, median OS was 42 months for patients with high anti-TA90 IgM titers (n = 28) vs. 9 months for patients with low titers (n = 36) (P = .0001). There was no significant correlation between total IgG/IgM and survival (P = .4107 and .4044, respectively). Multivariate analysis identified anti-TA90 IgM as the most significant independent variable influencing OS after complete resection of distant melanoma metastases (P = .0001). CONCLUSIONS: We conclude that the endogenous immune response to metastatic melanoma determines the outcome after surgical therapy. Enhancement of this specific immune response may prolong the survival of patients with distant melanoma metastases. PMID- 10791856 TI - The clinicopathological significance of preoperative serum-soluble interleukin-2 receptor concentrations in operable non-small-cell lung cancer patients. AB - BACKGROUND: Serum-soluble interleukin-2 receptor (IL-2R) concentrations have been found to be elevated in cancer patients. However, the importance of this finding in patients with non-small-cell lung cancer (NSCLC) has not been previously established. METHODS: Preoperative serum-soluble IL-2R concentrations were determined in 65 consecutive patients with operable NSCLC. The correlation of preoperative serum-soluble IL-2R concentrations with various clinicopathological features of this cancer was evaluated to clarify the clinical significance of this parameter. RESULTS: Although serum-soluble IL-2R concentrations were not significantly higher in operable NSCLC patients than in normal controls (P = .1180), serum-soluble IL-2R concentrations were significantly higher in patients with stage IIIB or IV disease than in normal controls (P = .0001). The presence of intrapulmonary metastasis was the only clinicopathological feature that was significantly correlated to serum-soluble IL-2R concentration (P = .0004). The sensitivity of serum-soluble IL-2R concentration in identifying the presence of intrapulmonary metastasis was 87.5%; specificity was 75%. CONCLUSIONS: Elevated preoperative serum-soluble IL-2R concentrations in patients with operable NSCLC reflect the occurrence of intrapulmonary metastasis. Preoperative examination of serum-soluble IL-2R concentrations may be valuable in the detection of the intrapulmonary metastasis preoperatively. PMID- 10791857 TI - Diagnostic accuracy of four approaches to interpreting neuropsychological test data. AB - The diagnostic accuracy of 4 approaches to interpreting neuropsychological test results are evaluated in 672 cognitively normal and 407 cognitively impaired persons using the Mayo Cognitive Factor Scales (G. E. Smith et al., 1994). The interpretation approaches studied are absolute scores, difference scores, profile variability, and change scores at 1- to 2-year test-retest intervals. All dependent measures were "highly significant" when diagnostic groups were compared on null hypothesis significance testing analyses. In contrast, varied accuracy rates were obtained when each measure's ability to correctly classify individuals was evaluated relative to overall diagnostic accuracy. Odds ratios were also highly varied and ranged from < or = 1.0 (i.e., chance) to 34.9. The clinical usefulness of absolute scores and difference scores in data interpretation is supported. Neither profile variability measures nor measures of change over time were diagnostically useful. PMID- 10791858 TI - Longitudinal changes in cognition, gait, and balance in abstinent and relapsed alcoholic men: relationships to changes in brain structure. AB - Chronic alcoholism is associated with cognitive and motor deficits, and there is evidence for reversibility with sobriety. Alcoholic men were examined after 1 month of sobriety and 2 to 12 months later with cognitive and motor tests and magnetic resonance imaging. In this naturalistic study, 20 alcoholic participants had abstained and 22 had resumed drinking at retesting. Abstainers sustained greater improvement than relapsers on tests of delayed recall of drawings, visuospatial function, attention, gait, and balance. Shrinkage in 3rd ventricle volume across all participants significantly correlated with improvement in nonverbal short-term memory. Additional brain structure-function relationships, most involving short-term memory, were observed when analyses were restricted to alcoholic men who had maintained complete abstinence, were light relapsers for at least 3 months, or had consumed no more than 10 drinks prior to follow-up testing. Thus, alcoholic men who maintain abstinence can show substantial functional improvement that is related to improvement in brain structure condition. PMID- 10791859 TI - Processing speed, working memory, and IQ: a developmental model of cognitive deficits following cranial radiation therapy. AB - IQ decrements following cranial radiation therapy (CRT) for acute lymphoblastic leukemia (ALL) are most apparent years after treatment. The authors examined a developmental model for delayed deficits by evaluating the relationship between processing speed, working memory, and IQ in long-term survivors of childhood ALL (n = 27) compared with demographically matched controls (n = 27). The ALL group treated with CRT showed deficits in IQ, working memory, and processing speed relative to controls. Differences in IQ between the CRT group and controls were mediated by differences in working memory. Processing speed did not fully account for the working memory deficit in the CRT group. Participants with ALL treated only with chemotherapy showed similar working memory and processing speed as matched controls. Data suggest that deficits in processing speed and working memory following CRT may underlie declines in IQ. PMID- 10791860 TI - Evaluation of a model of attention with confirmatory factor analysis. AB - Structural equation modeling (specifically, analysis of moment structures; J. L. Arbuckle, 1996) was used to evaluate the goodness of fit of a model of components of attention (A. F. Mirsky, B. J. Anthony, C. C. Duncan, M. B. Aheam, & S. G. Kellam, 1991) to neuropsychological test data from 2 samples. One sample consisted of psychiatrically normal persons with and without sleep-disordered breathing, and the other sample consisted of the adults studied by A. F. Mirsky et al. (1991), who gave rise to this model. That sample included psychiatric patients as well as normals. An exploratory data reduction procedure, principal components analysis, suggested that attention might be conceptualized as composed of 4 independent elements or components: focus-execute, sustain, shift, and encode. Neither the proposed orthogonal model nor a model permitting correlated factors adequately fit either data set, suggesting that these 4 attention constructs are as yet not clearly validated in the measures used to assess them. PMID- 10791861 TI - Development of psychometrically matched English and Spanish language neuropsychological tests for older persons. AB - Item response theory (IRT) methods were used to develop a neuropsychological test battery with matched English and Spanish language forms. Candidate items for 12 scales measuring core neuropsychological abilities were generated and administered to 200 community-dwelling elderly participants tested in Spanish and 208 tested in English. IRT methods were used to eliminate linguistically biased items and refine scales to assess broad ability ranges. Reasonably good psychometric matching of scales was achieved within and across English and Spanish language forms. All scales were sensitive to cognitive impairment as measured by the Mini-Mental State Examination (MMSE), with highly similar relationships between scale scores and MMSE across English and Spanish groups. The outcome supports the use of IRT methods in cross-cultural and multilingual test development and indicates that this strategy has potential for future neuropsychological test development. PMID- 10791862 TI - The cognitive correlates of white matter abnormalities in normal aging: a quantitative review. AB - Cerebral white matter of asymptomatic people frequently exhibits circumscribed areas of hyperintensity on magnetic resonance (MR) images and hypodensity on computed tomography scans. However, behavioral implications of this phenomenon remain unclear. In this meta-analysis, the authors examine cumulative evidence regarding the cognitive sequelae of white matter abnormalities in adults without dementia. The influence of potential moderator variables, such as neuroimaging technique, location of the lesions, rating scale, and demographic characteristics of the sample on the association between the burden of white matter hyperintensities and cognitive performance was also examined. Results indicate that white matter abnormalities observed on MR images are associated with attenuated performance on tasks of processing speed, immediate and delayed memory, executive functions, and indices of global cognitive functioning. There was no significant link between the white matter hyperintensities and psychometric indices of intelligence or fine motor performance. PMID- 10791863 TI - Recognition memory impairment in Parkinson's disease: power and meta-analyses. AB - Contrary findings notwithstanding, the prevailing notion is that recognition memory is little affected by Parkinson's disease (PD). Both a power analysis and a meta-analysis were conducted to help clarify the degree of recognition memory deficit associated with PD. The power analysis confirmed that, in general, memory studies of PD participants have been underpowered. This analysis indicated the need to pool study results in a subsequent meta-analysis, the main finding of which was that recognition memory deficits do occur with PD. The largest deficit occurs in PD participants with dementia. Nevertheless, deficits also occur in PD participants without dementia on medication, but nondopaminergic central nervous system abnormalities are more likely to underlie this deficit than PD medication itself. Future development of a theory of cognitive dysfunction in PD should take into account these recognition memory deficits, which may increase with disease progression. PMID- 10791864 TI - Working memory functioning in medicated Parkinson's disease patients and the effect of withdrawal of dopaminergic medication. AB - Cognitive impairments in Parkinson's disease (PD) could be explained by a central executive (CE) deficit in A. D. Baddeley's (1986) working memory model. To test this hypothesis, verbal, spatial, and double span tasks were given to 12 medicated PD patients and control participants, with varying recall delays. The double span task was assigned to explore the coordinating and integrating function of the CE, and lengthening the recall delay was expected to implicate more attentional resources. PD patients had lower spans relative to controls in all tasks. However, the more specific implication of the CE was difficult to prove. One reason could be that PD patients were on dopaminergic treatment when tested. To control this effect, 12 PD patients on and off medication were studied in a second experiment using the same tasks. PD patients off medication had lower spans only in the double task; this result underlines the role of dopamine on working memory processes. PMID- 10791865 TI - Responding to threat: hemispheric asymmetries and interhemispheric division of input. AB - This investigation examined how hemispheric asymmetry and interhemispheric processing contribute to attentional biases toward emotional information. Participants (n = 88) named the color of lateralized squares presented concurrently with neutral, positive, or threatening words. A left-hemisphere advantage in color naming was reduced when distractors were emotional, suggesting right-hemisphere priming by emotional stimuli. Furthermore, the advantage of dividing the word and color across visual fields was increased for emotion words when they were frequently presented, indicating a strategic use of interhemispheric division of labor to reduce the distracting effect of emotional words. Finally, participants with high levels of anxious apprehension were most likely to make use of this interhemispheric processing strategy, supporting a processing efficiency theory of cognitive function in anxiety. PMID- 10791866 TI - Extent, pattern, and correlates of remote memory impairment in Alzheimer's disease and Parkinson's disease. AB - Content and contextual memory for remote public figures and events was assessed with a modified version of the Presidents Test in patients with Alzheimer's disease (AD) or Parkinson's disease (PD). Contributions of executive functioning, semantic memory, and explicit anterograde memory to remote memory abilities were also examined. The AD group had temporally extensive deficits in content and contextual remote memory not accountable for by dementia severity. The PD group did not differ from the control group in remote memory, despite anterograde memory impairment. These results support the position that different component processes characterize remote memory, various mnemonic and nonmnemonic cognitive processes contribute to remote memory performance, and anterograde and remote memory processes are dissociable and differentially disrupted by neurodegenerative disease. PMID- 10791867 TI - When false recognition is unopposed by true recognition: gist-based memory distortion in Alzheimer's disease. AB - The authors examined false recognition of semantic associates in patients with probable Alzheimer's disease (AD), older adults, and young adults using a paradigm that provided rates of false recognition after single and multiple exposures to word lists. Using corrected false recognition scores to control for unrelated false alarms, the authors found that (a) the level of false recognition after a single list exposure was lower in AD patients than in controls; (b) across 5 trials, false recognition increased in AD patients, decreased in young adults, and showed a fluctuating pattern in older adults; and (c) all groups showed an increase in true recognition over the 5 trials. Analyses suggested that AD patients built up semantic gist across trials, whereas both control groups were able to use increased item-specific recollection and more conservative response criteria to suppress gist-based false alarms. PMID- 10791868 TI - Acetylcholine affects the spatial scale of attention: evidence from Alzheimer's disease. AB - Location precues were used to manipulate the spatial scale of attention in visual search for a target in an array of letters in patients with dementia of the Alzheimer type (DAT) and in age-matched older controls. Cue size varied in the amount of spatial precision conferred. Scopolamine, a muscarinic antagonist, decreased overall arousal and broadened spatial attention after a precise precue (small and valid) to target location for DAT patients but not for controls, suggesting a selective effect for attentional impairment induced by cholinergic blockade. In contrast, physostigmine, a cholinesterase inhibitor, did not alter the distribution of spatial attention relative to no-drug baseline testing for patients. Results support a differential role for cholinergic mechanisms in the modulation of the spatial scale of visual attention. PMID- 10791869 TI - Aging, episodic memory feeling-of-knowing, and frontal functioning. AB - Groups of normal old and young adults made episodic memory feeling-of-knowing (FOK) judgments and took 2 types of episodic memory tests (cued recall and recognition). Neuropsychological tests of executive and memory functions thought to respectively involve the frontal and medial temporal structures were also administered. Age differences were observed on the episodic memory measures and on all neuropsychological tests. Compared with young adults, older adults performed at chance level on FOK accuracy judgments. Partial correlations indicated that a composite measure of frontal functioning and FOK accuracy were closely related. Hierarchical regression analyses showed that the composite frontal functioning score accounted for a large proportion of the age-related variance in FOK accuracy. This finding supports the idea that the age-related decline in episodic memory FOK accuracy is mainly the result of executive or frontal limitations associated with aging. PMID- 10791870 TI - Intactness of inhibitory attentional mechanisms following severe closed-head injury. AB - Attentional problems are a common sequelae of closed-head injury (CHI). Research in the area of selective attention has pointed to the role of inhibitory mechanisms in the suppression of irrelevant information. In the current study, a negative priming paradigm was used to assess the inhibitory mechanisms of individuals suffering from a severe CHI. Twenty participants with severe CHIs (greater than 1 year postinjury) and 20 matched controls completed a negative priming task, as well as several other standardized tests of cognitive functioning. Within the negative priming task, 2 conditions were used to elicit information regarding facilitation by attended and ignored information and 1 condition was used to elicit inhibition of ignored information, as compared with a neutral control condition. Despite poorer performances on several tests of attention, there were no significant differences in the amount of inhibition displayed by the CHI participants as compared with the controls. Findings suggest that inhibitory processing deficits may not underlie the selective attention difficulties commonly seen following a severe CHI. PMID- 10791871 TI - Genetic subtypes of HIV type 1 in Republic of Congo. AB - To assess the molecular epidemiology of HIV-1 in Republic of Congo (Congo), we investigated 29 HIV-1s obtained from 82 Congolese AIDS and ARC patients in 1996 and 1997. Part of the env region including the V3 loop was phylogenetically analyzed. The genotypes observed were varied: of 29 specimens, 12 (41 %) were subtype A, 1 (3%) was subtype D, 6 (21%) were subtype G, 6 (21%) were subtype H, 2 (7%) were subtype J, and 2 (7%) could not be classified as any known subtypes (U, unclassified). The heterogeneous profile of HIV-1 infection was different from the profiles of neighboring Central African countries. These data show that subtypes G and H as well as subtype A were circulating with high prevalence. The fact that new genetic subtypes (J and U) are circulating indicates a need for a greater surveillance for these subtypes both in Congo as well as in other parts of the world. PMID- 10791872 TI - An improved algorithm for determining HIV type 1 subtypes in a primary laboratory in Uganda. AB - A pilot study was undertaken with the objective of developing a simple, economical, and efficient algorithm through which to subtype HIV-1 in a large epidemiological cohort study in Uganda. A peptide enzyme immunoassay (PEIA) employing both V3 and gp41 regions and a heteroduplex mobility assay (HMA) were evaluated in comparison with DNA sequencing. Of 146 samples selected, 115 (79%) were successfully sequenced. Taking sequence data as the "gold standard," other assays were compared with these data. The HMA correctly identified 95 (83%) of the samples, and only 1 sample was wrongly identified. The V3 PEIA alone and in combination with gp41 peptides correctly identified 76 and 78% of the samples, respectively; however, the number of wrongly identified samples was four times less with the combination compared with V3 peptides alone (4 versus 16%). The sensitivity, specificity, and positive and negative predictive values for serotype A and D samples were greater for the combination than V3 peptides alone. We have described a new algorithm to segregate subtypes A and D. This algorithm uses the two peptide assays followed by HMA and then DNA sequencing for untypable samples, giving an accuracy of 95% at a cost of 37 and 21% for consumables compared with subtyping all the samples by HMA or DNA sequencing, respectively. This proposed approach is suitable for epidemiological studies in Uganda and other regions with a predominance of A and D subtypes. PMID- 10791873 TI - Anti-human immunodeficiency virus activity of novel aminoglycoside-arginine conjugates at early stages of infection. AB - Conjugates of L-arginine with aminoglycosides have already been described as potent in vitro inhibitors of the HIV-1 Tat-trans-activation responsive element interaction. The polycationic nature of these agents leads us to suggest that they may be active against HIV-1 replication by inhibiting earlier stages of the virus life cycle. We have found that R4K and R3G, kanamycin A, and gentamicin C, conjugated with arginine, inhibited HIV-1 NL4-3 replication at EC50 values of 15 and 30 microM for R3G and R4K, respectively, without a detectable tonic effect on MT-4 cells at concentrations higher than 4000 and about 1000 microM, respectively. Both compounds inhibited the binding of a monoclonal antibody (12G5) directed to CXCR4 as well as the intracellular Ca2+ signal induced by the chemokine SDF-1alpha on CXCR4+ cells, suggesting that aminoglycoside-arginine conjugates interact with CXCR4, the coreceptor used by T-tropic, X4 strains of HIV-1. On the other hand, CB4K, a conjugate of kanamycin A with gamma guanidinobutyric acid, structurally similar to R4K, failed to display any anti HIV activity of CXCR4 antagonist activity. An HIV-1 strain that was made resistant to the known CXCR4 antagonist AMD3100 was cross-resistant to both R4K and R3G. However, unlike SDF-1alpha and R4K, R3G inhibited the binding of HIV-1 to MT-4 cells. Aminoglycoside-arginine conjugates inhibit HIV replication by interrupting the early phase of the virus life cycle, namely virus binding to CD4 cells and interaction with CXCR4. R3G and R4K may serve as prototypes of novel anti-HIV agents and should be further studied. PMID- 10791874 TI - Fetal mitochondrial heart and skeletal muscle damage in Erythrocebus patas monkeys exposed in utero to 3'-azido-3'-deoxythymidine. AB - 3'-azido-3'-deoxythymidine (AZT) is given to pregnant women positive for the human immunodeficiency virus type 1 (HIV-1) to reduce maternal-fetal viral transmission. To explore fetal mitochondrial consequences of this exposure, pregnant Erythrocebus patas monkeys were given daily doses of 1.5 mg (21% of the human daily dose) and 6.0 mg (86% of the human daily dose) of AZT/kg body weight (bw), for the second half of gestation. At term, electron microscopy of fetal cardiac and skeletal muscle showed abnormal and disrupted sarcomeres with myofibrillar loss. Some abnormally shaped mitochondria with disrupted cristae were observed in skeletal muscle myocytes. Oxidative phosphorylation (OXPHOS) enzyme assays showed dose-dependent alterations. At the human-equivalent dose of AZT (6 mg of AZT/kg bw), there was an approximately 85% decrease in the specific activity of NADH dehydrogenase (complex I) and three- to sixfold increases in specific activities of succinate dehydrogenase (complex II) and cytochrome-c oxidase (complex IV). Furthermore, a dose-dependent depletion of mitochondrial DNA levels was observed in both tissues. The data demonstrate that transplacental AZT exposure causes cardiac and skeletal muscle mitochondrial myopathy in the patas monkey fetus. PMID- 10791875 TI - Both serum HIV type 1 RNA levels and CD4+ lymphocyte counts predict clinical outcome in HIV type 1-infected subjects with 200 to 500 CD4+ cells per cubic millimeter. AIDS Clinical Trials Group Study 175 Virology Study Team. AB - To evaluate HIV-1 RNA and CD4+ cell responses to therapy as predictors of clinical progression and to evaluate levels and trends of these markers prior to clinical failure, HIV-1 RNA measurements were retrospectively obtained on subjects who progressed to AIDS or death and a random sample of subjects who did not. Samples were taken from AIDS Clinical Trials Group Study 175, a randomized trial comparing nucleoside analog therapies in subjects with CD4+ cell counts of between 200 and 500 cells/mm3. HIV-1 RNA and CD4+ cell count independently predicted clinical progression. Risk of subsequent progression is best captured by the change to the last measured value for CD4+ cell count and the area under the curve minus baseline, a measure of viral replication over time, for HIV-1 RNA. Subjects who failed had lower CD4+ cell counts, greater rates of CD4+ cell decline, and higher HIV-1 RNA levels, but not greater rates of HIV-1 RNA increase than subjects who did not. Subjects who maintained more than 200 CD4+ cells/mm3 and fewer than 10,000 copies of HIV-1 RNA per milliliter had low risk of progression. During the first few months of therapy, treatments are best monitored by regular HIV-1 RNA and less frequent CD4+ cell measurements. Thereafter, both markers should be monitored on a similar schedule to identify rapidly declining CD4+ cell counts, or adverse levels of either. These results further delineate the prognostic significance of HIV-1 RNA and CD4+ cell count and should help to better define their utility in the practice setting. PMID- 10791876 TI - AIDSVAX (MN) in Bangkok injecting drug users: a report on safety and immunogenicity, including macrophage-tropic virus neutralization. AB - A randomized, double-blind, placebo-controlled phase I/II study of AIDSVAX (MN) was conducted among injecting drug users in Bangkok, Thailand. Four doses of vaccine (300 microg of MN-rgp120 in alum) or placebo (alum) were given at study entry and at 1, 6, and 12 months. The objectives of the study were to evaluate (1) the feasibility of conducting vaccine trials in this population; (2) the safety of this candidate AIDS vaccine; and (3) the immunogenicity of this vaccine. Thirty-three volunteers (22 vaccine and 11 placebo recipients) were recruited. None were lost to follow-up during the 18-month study. Mild reactogenicity was noted, which was similar in both vaccine and placebo recipients. The vaccine induced anti-HIV-1 antibody in all vaccine recipients. Maximal titers of binding antibodies of MN-rgp120 and the V3 domain of MN-rgp120 were induced after the third (6 month) dose while maximal neutralizing antibodies followed the fourth (12 month) dose. The vaccine-induced antibodies from several volunteers were capable of neutralizing macrophage-tropic, subtype B viruses (301660 and JRCSF) detected in a PBMC-based assay. Binding and neutralizing antibodies declined about 10-fold in the 6 months after the last boost. Two vaccinees became infected during the trial, both with subtype E viruses. A phase III efficacy trial, using a bivalent gp120 vaccine containing antigens from a subtype B virus (MN) and a subtype E virus (A244), was initiated in March 1999 in injecting drug users in Bangkok. PMID- 10791877 TI - Real-time polymerase chain reaction assay for cell-associated HTLV type I DNA viral load. AB - We have developed a quantitative real-time PCR assay for HTLV-I DNA. This assay approach uses real-time monitoring of fluorescent signal generation as a consequence of Taq-mediated amplification of specific target sequences to allow real-time kinetic analysis of amplicon production. This kinetic approach yields excellent sensitivity and an extremely broad linear dynamic range, and ensures that quantitation is based on analysis during the exponential phase of amplification, regardless of the input template copy number. The HTLV-I DNA assay has a nominal threshold sensitivity of 10 copy Eq/reaction, although single-copy plasmid template can be detected at frequencies consistent with statistical prediction. The linear dynamic range is in excess of 5 logs. Interassay reproducibility averages 14% (coefficient of variation) for control templates over a range of 10(1) to 10(6) copy Eq/reaction and 25%, based on studies of extraction and analysis of replicate aliquots of PBMC specimens from HTLV-I infected subjects. The primer/probe combination targets tax sequences conserved across described HTLV-I and HTLV-II isolates. Parallel quantitation in the same samples of an endogenous sequence present at a known copy number per cell allows normalization of results for potential variation in DNA recovery. Availability of this assay should facilitate studies of basic pathogenesis and clinical evaluation of HTLV-I and HTLV-II infection, as well as assessment of therapeutic approaches. PMID- 10791878 TI - Feline immunodeficiency virus clade C mucosal transmission and disease courses. AB - The transmissibility and pathogenicity of a clade C feline immunodeficiency virus (FIV-C) was examined via the oral-nasal, vaginal, or rectal mucosa. FIV-C was transmissible by all three mucosal routes. Vaginal transmission was most efficient (100%), oral exposure resulted in a 80% infection rate, and rectal transmission was least effective (44%). In contrast to previous intravenous passage studies, a broader range of host-virus relationships was observed after mucosal exposure. Three categories of FIV-C infection were defined: (1) rapidly progressive infection marked by high virus burdens and rapid CD4+ cell depletion (43% of vaginally exposed animals); (2) conventional (typical) infection featuring slowly progressive CD4+ cell decline (61% of all exposed animals); and (3) regressive (transient) infection marked by low and then barely detectable virus burdens and no CD4+ cell alterations (22% of rectally inoculated cats). These disease courses appear to have parallels in mucosal HIV and SIV infections, emphasizing the importance of the virus-mucosa interface in lentiviral pathogenesis. PMID- 10791879 TI - Early activation and proliferation of T cells in simian immunodeficiency virus infected rhesus monkeys. AB - To longitudinally determine T cell activation and turnover in early simian immunodeficiency virus (SIV) infection of macaques, immunological and virological parameters were monitored in 10 SIV-infected animals starting before infection until 40 weeks postinfection (wpi). Lymphocyte subsets in blood and lymph nodes (LNs) were characterized by three-color flow cytometry for expression of markers of activation, proliferation, and differentiation. As early as 1 wpi, CD69 expression was upregulated both on CD4+ and CD8+ T cells, indicative of an early activation of these cells. Whereas this activation led to increased proliferation, determined by expression of Ki-67, and absolute numbers of CD8+ T cells, CD4+ T cells showed a decreased expression of Ki-67 and reduced counts in blood at 2 wpi. Later, the percentage of Ki-67-expressing CD4+ T cells in blood and LNs increased again above preinfection levels in most animals but remained low in two monkeys progressing to AIDS. These findings suggest that T cells are activated after SIV infection, leading to increased T cell proliferation already in the early asymptomatic phase. In addition, we found a correlation between the capacity to regenerate CD4+ T cells by peripheral proliferation and the disease course. Moreover, our data indicate that the increased peripheral T cell proliferation during immunodeficiency virus infection is probably not caused by the effort of the immune system to maintain T cell homeostasis but may be a reflection of the ongoing immune response against the virus. PMID- 10791880 TI - The Kell blood group system: Kell and XK membrane proteins. AB - Two membrane proteins express the antigens that comprise the Kell blood group system. A single antigen, Kx, is carried on XK, a 440-amino acid protein that spans the membrane 10 times, and more than 20 antigens reside on Kell, a 93-kd, type II glycoprotein. XK and Kell are linked, close to the membrane surface, by a single disulfide bond between Kell cysteine 72 and XK cysteine 347. Although primarily expressed in erythroid tissues, Kell and XK are also present in many other tissues. The polymorphic forms of Kell are due to single base mutations that encode different amino acids. Some Kell antigens are highly immunogenic and may cause strong reactions if mismatched blood is transfused and severe fetal anemia in sensitized mothers. Antibodies to KEL1 may suppress erythropoiesis at the progenitor level, leading to fetal anemia. The cellular functions of Kell/XK are complex. Absence of XK, the McLeod phenotype, is associated with acanthocytic red blood cells (RBCs), and with late-onset forms of muscular dystrophy and nerve abnormalities. Kell, by homology, is a member of the neprilysin (M13) family of membrane zinc endopeptidases and it preferentially activates endothelin-3 by specific cleavage of the Trp21-Ile22 bond of big endothelin-3. PMID- 10791881 TI - The Duffy protein: a malarial and chemokine receptor. AB - A major advance towards understanding the Duffy blood group system has been achieved with the cloning of FY, a single-copy gene located in the 1q22->q23 region of chromosome 1. The product of FY Is an acidic glycoprotein (gp-Fy), which spans the plasma membrane seven times and has an exocellular N-terminal domain and an endocellular C-terminal domain. The system consists of four alleles, five phenotypes, and five antigens. FYA, FYB, FYB(ES), and FYB(WK) are the alleles; Fy(a+b-), Fy(a-b+), Fy(a+b+), Fy(a-b+(wK)), and Fy(a-b-), are the phenotypes, and Fy(a), Fy(b), Fy3, Fy5, and Fy6 are the antigens. Fy(a-b-), or Duffy-negative individuals, lack the Duffy protein on erythrocytes and are predominantly African and American blacks. They have the FYB(Es) allele with a mutation in the promoter region, which abolishes the expression of the protein in erythrocytes only. In the few cases of non-black Fy(a-b-) individuals, a nonsense mutation prevents the synthesis of gp-Fy. In Fy(a-b+(wk)) erythrocytes, the Fy(b) antigen is weakly expressed due to a reduced amount of the protein. The Fy5 antigen includes the Rh protein, and the Fy6 antigen is defined by a murine monoclonal antibody. Gp-Fy is produced in several cell types, including endothellal cells of capillary and postcapillary venules, epithelial cells of kidney collecting ducts, and lung alveoli, as well as PurkinJe cells of the cerebellum. The Duffy protein plays a role in inflammation and in malaria Infection. The protein is a member of the superfamily of chemokine receptors and is the receptor to which certain malarial parasites bind to invade red blood cells. The parasite-specific binding site, the binding site of chemokines, and the major antigenic domains are located in overlapping regions at the exocellular N terminus of the Duffy protein. PMID- 10791883 TI - Introduction to porcine red blood cells: implications for xenotransfusion. AB - Advances in the field of xenotransplantation raise the intriguing possibility of using porcine red blood cells (pRBCs) as an alternative source for blood transfusion. The domestic pig is considered the most likely donor species for xenotransplantation. However, identification of xenoantigens on porcine erythrocytes and elucidation of their possible roles in antibody-mediated RBC destruction are necessary for developing clinical strategies to circumvent immunological incompatibility between humans and pigs. Although the alphaGal epitope (Galalpha1,3Galbeta1,4GIcNAc-R) is the major xenoantigen on porcine erythrocytes and is responsible for the binding of the majority of human natural antibodies, other non-alphaGal xenoantigens have been identified. The importance of these non-alphaGal xenoantigens in binding human natural antibodies and subsequently triggering immunological responses cannot be underestimated. Our data suggest that non-alphaGal xenoantigen(s) identified on the porcine erythrocyte membrane are not only recognized by xenoreactive human natural antibodies but are also involved in complement-mediated hemolysis. PMID- 10791882 TI - Red blood cell surface adhesion molecules: their possible roles in normal human physiology and disease. AB - Human erythrocytes express a relatively large number of known adhesion receptors, despite the fact that red blood cells (RBCs) are generally considered to be nonadhesive for endothelial cell surfaces. Some of these adhesion receptors are expressed by many other tissues, while others have more limited tissue distribution. Some adhesion receptors, including CD36 and VLA-4, are only expressed by immature erythroid cells, while others are present on mature erythrocytes. The structure and function of these proteins is reviewed here. LW, CD36, CD58, and CD147 have been shown in other tissues to mediate cell-cell interaction. Other receptors, such as CD44, VLA-4, and B-CAM/LU, can mediate adhesion to components of extracellular matrix. In addition, their roles in normal erythropolesis, as well as in the pathophysiology of human disease, are summarized. The most convincing evidence for a pathophysiologic role for any of these receptors on erythrocytes comes from studies of cells from patients homozygous for hemoglobin S, as RBC adhesion is thought to contribute to vaso occlusion. Thus, receptors such as B-CAM/LU may become targets for future therapy aimed at preventing or ameliorating this thrombotic process. PMID- 10791884 TI - Molecular biology and genetics of the Rh blood group system. AB - The Rh (Rhesus) blood group system is the most complex of the known human blood group polymorphisms. The expression of its antigens is controlled by a two component genetic system consisting of RH and RHAG loci, which encode Rh30 polypeptides and Rh50 glycoprotein, respectively. Over the past decade, there has been a rapid advance in knowledge of the biochemistry, molecular biology, and genetics of the Rh genes and proteins. The primary structures of D and CcEe antigens have become well understood and the molecular genetic basis of a vast array of phenotype polymorphisms has been delineated. The identification of various molecular defects associated with Rh deficiency syndrome clarifies the nature of the amorph, suppressor, and modifier genes. The observed mutation spectrum defines a basic set of components essential for Rh complex assembly in the erythrocyte membrane. The resulting molecular information, combined with new experimental tools, is helping to dissect the fine structure of Rh antigens in terms of epitope mapping. The discovery of novel Rh homologs in primitive organisms and in nonerythroid tissues opens new avenues of research beyond the scope of erythrocytes and Rh antigens. This review provides an update on the Rh family in antigen expression, phenotype diversity, and disease association. PMID- 10791886 TI - The role of blood group antigens in infectious diseases. AB - The medical literature contains a large number of publications attempting to correlate blood groups with disease. Many of these reports are poorly documented and have limited scientific validity. Only a few agents, such as malaria parasites and parvovirus B19, infect red blood cells (RBCs) and precursors. Most other agents use RBCs as carriers to the target tissue. There is an excess of blood group A individuals among cancer patients compared with normal individuals; malignancy has also been associated with the Lewis antigen. Plasmodium vivax only enters RBCs when the Fy6 Duffy protein is present. Certain Escherichia coli organisms will only attach to epithelial cells carrying P or Dr blood group antigens. The P antigen Is also the receptor for parvovirus B19. Le(b) appears to be the receptor for Helicobacter pylori in gastric tissue. The high frequency blood group antigen AnWJ is the receptor for Haemophilus influenzae. Knowledge of the functions of RBC surface molecules Is expanding and the ability to generate experimental animals devoid of certain molecules will clarify their physiological role. PMID- 10791885 TI - Applications of molecular biology techniques to transfusion medicine. AB - Other articles in this issue of Seminars in Hematology have reviewed the results of basic research in relation to the understanding of the genes, the molecular basis of blood group variants, and structural and functional aspects of the proteins carrying blood group antigens. Although molecular techniques are currently being used in a limited fashion in clinical laboratories, their application has far-reaching possibilities and undoubtedly will be soon applied more generally. We focus on two general areas: molecular genotyping for blood group antigens and their expression analysis in heterologous systems. PMID- 10791887 TI - Red blood cell polyagglutination: clinical aspects. AB - Polyagglutination is the term applied to red blood cells (RBCs) that are agglutinated by almost all samples of human sera from adults but not by autologous serum or sera of newborns. The polyagglutinable state may be transient or persistent. Transient polyagglutinability results from the exposure of normally cryptic antigens by bacterial enzymatic activity during the course of an infectious process. RBCs are polyagglutinable because most sera from adults contain agglutinins for the exposed antigens. This type of polyagglutination can often be reproduced in vitro with bacterial culture fluids or isolated enzymes. Persistent polyagglutination may be a consequence of somatic mutation leading to a cellular lineage characterized by an enzyme deficiency that results in exposure of a normally cryptic antigen, Tn. Most human sera contain anti-Tn. Tn polyagglutination is regularly accompanied by leukopenia and thrombocytopenia and has been associated with leukemia. Other forms of persistent polyagglutination are due to the inheritance of rare blood groups or are associated with a hematologic dyscrasia. PMID- 10791888 TI - Summary of the clinical significance of blood group alloantibodies. AB - The knowledge of the potential clinical significance of alloantibodies to blood group antigens is essential for selecting appropriate red blood cell (RBC) products for transfusion. This article provides a summary of clinical and technical aspects of many alloantibodies. PMID- 10791889 TI - Myosin II-independent cytokinesis in Dictyostelium: its mechanism and implications. AB - Similar to higher animal cells, ameba cells of the cellular slime mold Dictyostelium discoideum form contractile rings containing filaments of myosin II during mitosis, and it is generally believed that contraction of these rings bisects the cells both on substrates and in suspension. In suspension, mutant cells lacking the single myosin II heavy chain gene cannot carry out cytokinesis, become large and multinucleate, and eventually lyze, supporting the idea that myosin II plays critical roles in cytokinesis. These mutant cells are however viable on substrates. Detailed analyses of these mutant cells on substrates revealed that, in addition to "classic" cytokinesis which depends on myosin II ("cytokinesis A"), Dictyostelium has two distinct, novel methods of cytokinesis, 1) attachment-assisted mitotic cleavage employed by myosin II null cells on substrates ("cytokinesis B"), and 2) cytofission, a cell cycle-independent division of adherent cells ("cytokinesis C"). Cytokinesis A, B, and C lose their function and demand fewer protein factors in this order. Cytokinesis B is of particular importance for future studies. Similar to cytokinesis A, cytokinesis B involves formation of a cleavage furrow in the equatorial region, and it may be a primitive but basic mechanism of efficiently bisecting a cell in a cell cycle coupled manner. Analysis of large, multinucleate myosin II null cells suggested that interactions between astral microtubules and cortices positively induce polar protrusive activities in telophase. A model is proposed to explain how such polar activities drive cytokinesis B, and how cytokinesis B is coordinated with cytokinesis A in wild type cells. PMID- 10791890 TI - Overexpression of PRA2, a Rab/Ypt-family small GTPase from Pea Pisum sativum, aggravates the growth defect of yeast ypt mutants. AB - A large number of Rab/Ypt-family small GTPases have been identified from higher plants. While some of them can complement yeast ypt mutants, the expression of Arabidopsis Ara4 protein aggravated the growth defect of a subset of ypt mutants, probably because of the titration of common regulator(s) of yeast Ypt proteins [Ueda, T. et al. (1996) Plant Cell, 8: 2079-20911. PRA2 from pea Pisum sativum encodes an interesting Rab GTPase whose expression is regulated by light [Yoshida, K. et al. (1993) Proc. Natl. Acad. Sci. USA, 90: 6636-6640]. We examined whether PRA2 complements any of the yeast ypt mutants and found again that PRA2 does not complement but rather confers the growth defect to some of the ypt mutants. No growth defect was observed when PRA2 was expressed in the wild type yeast cells. Unlike the case of Ara4, neither Arabidopsis nor yeast GDI remedied the growth defect by Pra2, indicating that the mechanism of the exacerbation is different. Mutational analysis of PRA2 suggests that the growth inhibition can be ascribed to unidentified factor(s) which prefers the GTP-bound form of Pra2. This yeast system will be useful for identifying such putative regulatory factor(s) from yeast and plants and analyzing their interactions with Pra2. PMID- 10791891 TI - Murine Delta homologue, mDelta1, expressed on feeder cells controls cellular differentiation. AB - The Delta/Serrate-Notch pathway is involved in intercellular signaling that controls cell fate during the development of invertebrates and vertebrates. Delta is a prototype of Notch ligands and has been studied extensively in Drosophila. In higher vertebrates, four Delta/Serrate homologues and four Notch homologues have been identified. Recent studies showed that the murine Delta homologue, mDelta1, is essential in early embryogenesis. The biological activity of mammalian Delta and its roles in cellular differentiation, however, have remained unclear. In this study, we first surveyed expression of mDelta1 in the adult mouse and found it to be present in a wide range of tissues. For testing biological activity of mDelta1, we expressed a mDelta1 full-length cDNA in L cells using a eukaryotic expression vector. Effects of mDelta1 on cellular differentiation were examined in two independent systems, featuring C2C12 myogenic differentiation and multipotent murine bone marrow cell differentiation. Inhibition of the former was observed with mDelta1 expression on L cells, associated with suppression of myogenin, a myogenic transcription factor. Expression of mDelta1 in conjunction with GM-CSF promoted differentiation of bone marrow cells to myeloid dendritic cells at the expense of other lineages. Although the effects of mDelta1 on two differentiation systems appeared opposing, as inhibition occurring in one and induction in the other, this can be understood by the unifying concept of generation of diverse cell types from equivalent progenitors. Thus, the present study provided evidence that mammalian Delta participates in intercellular signaling, determining the cell fate in a wide variety of tissues. PMID- 10791892 TI - Ser787 in the proline-rich region of human MAP4 is a critical phosphorylation site that reduces its activity to promote tubulin polymerization. AB - p34cdc2 kinase-phosphorylation sites in the microtubule (MT)-binding region of MAP4 were determined by peptide sequence of phosphorylated MTB3, a fragment containing the carboxy-terminal half of human MAP4. In addition to two phosphopeptides containing Ser696 and Ser787 which were previously indicated to be in vivo phosphorylation sites, two novel phosphopeptides, containing Thr892 or Thr901 and Thr917 as possible phosphorylation sites, were isolated, though only in in vitro phosphorylation. The role of phosphorylation at Ser696 and Ser787, which were differently phosphorylated during the cell cycle (Ookata et al., (1997). Biochemistry, 36: 15873-15883), was investigated in MT-polymerization, using MAP4 Ser to Glu mutants, which mimic phosphorylation at each site. Mutation of Ser787 to Glu strikingly reduced the MAP4's MT-polymerization activity, while Glu-mutation at Ser696 did not. These results suggest that Ser787 could be the critical phosphorylation site causing MTs to be dynamic at mitosis. PMID- 10791893 TI - Different responses of polyploidized V79 cells after removal of two drugs, demecolcine and K-252a. AB - To examine whether or not cells polyploidized by different mechanisms behave in a different manner after drug removal, V79 Chinese hamster cells were assessed by flow cytometry (FCM) after their polyploidization by demecolcine and K-252a, inhibitors of spindle-fiber formation and protein kinase, respectively. Cell cycle analysis of DNA histograms of V79 cells before and after the drug release was performed. With both drugs, the ploidy of V79 cells increased just after the drug removal and was maintained for a week. A difference was evident 10 days after the release. Tetraploid cells were the main population from 10 to 18 days after the release of K-252a, but not demecolcine. Cell cycle parameters were almost the same in pseudo diploid and tetraploid V79 cells, except for the tetraploid S phase which was 2h longer. PMID- 10791894 TI - Identification and characterization of two flavohemoglobin genes in Dictyostelium discoideum. AB - Flavohemoglobins are being identified in an expanding number of prokaryotes and unicellular eukaryotes. These molecules consist of an N-terminal hemoglobin domain and a C-terminal oxidoreductase domain, and are considered to function in storage or as sensors for O2, and in defense against oxidative stress and/or NO. However, their physiological significance has not yet been determined. Here, we isolated and analyzed two flavohemoglobin genes of Dictyostelium discoideum, DdFHa and DdFHb, which lie close to each other in the genome. DdFHs were induced by submerged conditions, and enriched in the sexually mature cells of D. discoideum. Although they were not essential for growth or development under standard laboratory conditions, disruption of both genes caused an increase in number of large but uninuclear cells, and hypersensitivity to higher concentrations of glucose and to NO releasers. These results indicate that DdFHs are responsible for transducing NO signals to maintain normal cellular conditions against environmental stresses. PMID- 10791895 TI - Artificial phosphorylation removes Gelsolin's dependence on calcium. AB - Gelsolin is one of the best known actin-binding proteins with several distinct activities regulated by calcium. Using a kinase fraction isolated from mitotic HeLa cells, we found that the plasma form of gelsolin can be phosphorylated at a site located within the NH2-terminus region which does not exist in the cytoplasmic form. After this phosphorylation, gelsolin no longer requires Ca2+ for activity; it severs and subsequently caps actin filaments, and nucleates filament formation in Ca2+-free solution. These findings may clarify the mechanism of gelsolin regulation by Ca2+, and indicate that changes in electrical interactions between the NH2- and COOH-terminal ends are important for this regulation. Moreover, since only a single site is phosphorylated, and since the phosphorylated region does not contribute to this protein's own activity, the results suggest that a single chemical charge modification at a site away from the protein's core structure, such as this phosphorylation site, is sufficient to alter the protein's function. PMID- 10791896 TI - Ex vivo purging and cell expansion. PMID- 10791897 TI - Investigator profile. PMID- 10791898 TI - Implications for Src kinases in hematopoiesis: signal transduction therapeutics. AB - Signal transduction therapeutics is now the dominant theme of drug discovery, and its most immediate impact will be in cancer therapeutics. Blood cell proliferation, differentiation, and activation are controlled by cytokines, whose receptors contain tyrosine kinase catalytic domains or recruit cytosolic tyrosine kinases. Among the most important cytosolic protein tyrosine kinases are the Src and Jak families. Receptor or cytosolic protein tyrosine kinases activate a similar set of intracellular signaling molecules. In blood cells, excessive tyrosine kinase activity is associated with either cancer or autoreactive diseases. Therefore, tyrosine kinases and their substrates serve as excellent candidates for drug intervention. Herceptin has been approved for use in breast cancer. Other agents, such as SU101 and CGP 57418B, are well into phase I-III trials. Newer, more selective tyrosine kinase inhibitors are being evaluated for future use in the treatment of hematologic and solid tumors as well as a wide range of inflammatory or autoimmune diseases. PMID- 10791899 TI - The removal of human breast cancer cells from hematopoietic CD34+ stem cells by dielectrophoretic field-flow-fractionation. AB - Dielectrophoretic field-flow-fractionation (DEP-FFF) was used to purge human breast cancer MDA-435 cells from hematopoietic CD34+ stem cells. An array of interdigitated microelectrodes lining the bottom surface of a thin chamber was used to generate dielectrophoretic forces that levitated the cell mixture in a fluid flow profile. CD34+ stem cells were levitated higher, were carried faster by the fluid flow, and exited the separation chamber earlier than the cancer cells. Using on-line flow cytometry, efficient separation of the cell mixture was observed in less than 12 min, and CD34+ stem cell fractions with a purity >99.2% were obtained. The method of DEP-FFF is potentially applicable to many biomedical cell separation problems, including microfluidic-scale diagnosis and preparative scale purification of cell subpopulations. PMID- 10791900 TI - Distinct effect of retroviral-mediated IFN-alpha gene transfer on human erythroleukemic and CD34+ cell growth and differentiation. AB - Human interferon-alpha (IFN-alpha) has been used in the management of leukemia, but its diverse adverse effects may influence the ability of IFN-alpha to treat this disease. We constructed two retroviral vectors, LSN-IFN-alpha and LNC-IFN alpha, in which IFN-alpha cDNA was driven by viral LTR and CMV promoters, respectively. After transduction into the PA317 and PG13 retroviral packaging cells, high titers of retrovirus were produced and were used to infect K562 and human BM CD34+ hematopoietic cells. The IFN-alpha gene expression in transduced K562 cells was confirmed by Northern blot, RT-PCR, RIA, and biologic assay. Cell proliferation and cell viability in IFN-alpha-transduced K562 cells were significantly suppressed as compared with control K562 cells. Although the IFN alpha expression in K562 cells did not affect BCR/ABL expression, it apparently upregulated the production of adhesion molecules (VLA-4 and Mac-1). We evaluated the effect of IFN-alpha gene transfer on human CD34+ cells infected with LSN-IFN alpha retrovirus with the aid of fibronectin (FN) fragment CH-296 and growth factors. RIA showed that IFN-alpha-transduced CD34+ cells produced 72.2+/-15 U/ml of IFN-alpha compared with 4.3+/-1.2 U/ml in control CD34+ cells. Methylcellulose clonogenic assay indicated that IFN-alpha-transduced CD34+ cells produced similar numbers of burst-forming units-erythrocytes (BFU-E)/colony-forming units-GM (CFU GM) colonies as compared with control CD34+ cells. Selected colonies expressed IFN-alpha and neo(r) mRNA, as measured by RT-PCR. These studies indicate that retrovirus-mediated IFN-alpha gene transfer may provide a useful tool for studying the effect of IFN-alpha gene transfer on leukemic cells and long-lived CD34+ cells. PMID- 10791901 TI - Retroviral-mediated transfer and expression of the multidrug resistance protein 1 gene (MRP1) protect human hematopoietic cells from antineoplastic drugs. AB - Multidrug resistance protein (MRP1) is a member of the ATP-binding cassette (ABC) transmembrane transporter superfamily that confers multidrug resistance. The transfer and expression of the MRP1 gene in human hematopoietic stem cells may be a useful alternative to multidrug resistance (MDR1) gene transfer for protection from the myelosuppressive effects of chemotherapy in cancer patients. We constructed a gibbon ape leukemia virus packaging cell line (PG13) using the human MRP1 cDNA in a Moloney murine leukemia virus (MoMuLV) backbone containing a modified LTR. This PG13-based cell line, designated MRP1-PG13, produces retroviral vectors bearing the MRP1 gene at a titer of 1.7x10(5) viral particles/ml. Transduction of the human leukemic cell line K562 showed that viral MRP1-PG13 supernatants routinely transfer the MRP1 gene to approximately 35% of target K562 cells, of which at least one third are capable of proliferating in the presence of otherwise toxic concentrations of etoposide. Southern blot analyses indicated that most clones had only one proviral integration. Northern blot analysis of expanded K562 clones showed the presence of a major full-length approximately 8-kb MRP1 transcript as well as a minor approximately 6-kb transcript in all clones. Flow cytometric analysis of the producer cells and clones of transduced K562 cells demonstrated significantly increased MRP1 expression in these cells (approximately 30-fold increase). Human bone marrow mononuclear cells and CD34+ cells were also transduced with MRP1-PG13 supernatants on fibronectin-coated culture flasks in the presence of SCF, IL-3, and IL-6. PCR analysis of individual hematopoietic colonies in methylcellulose cultures demonstrated proviral DNA in approximately 10% of unselected human hematopoietic progenitor cells cultured from nonsorted mononuclear cell samples and in up to approximately 75% of progenitors when CD34-enriched cell populations were targeted. To assess functional MRP1 gene expression, normal human hematopoietic progenitors and K562 cells were cultured in methylcellulose assays containing vincristine or etoposide. All transduced samples gave rise to approximately 10% drug-resistant colonies, which were shown to be provirus positive by PCR. Our studies document the development of an amphotropic MRP1 retroviral vector producer cell line and pave the way for large animal and preclinical studies of chemoprotection by MRP1 gene transfer. PMID- 10791902 TI - Human mesenchymal stem cells provide stromal support for efficient CD34+ transduction. AB - Human mesenchymal stem cells (hMSC)-nonhematopoietic cells within the bone marrow microenvironment that can be culture expanded to a uniform population of fibroblastic cells-have been shown to support long-term hematopoiesis of CD34+ cells. Because direct contact between stromal elements and CD34+ cells enhances long-term engraftment, we postulated that hMSC would be a good alternative to the more heterogeneous stroma currently used in gene transfer studies. We used hMSC to support retroviral gene transfer of the G156A MGMT (deltaMGMT) gene encoding an alkyltransferase (AGT), which confers drug resistance to a combination of O6 benzylguanine (BG) plus the alkylating agents BCNU and temozolomide (TMZ) in human hematopoietic progenitors. In the presence of IL-3, IL-6, SCF, or leukemia inhibitory factor (LIF) and Flt-3 ligand, hMSC facilitated expansion and retroviral transduction of human peripheral blood-mobilized CD34+ cells. Furthermore, the transduced cells expressed AGT in 29% of hematopoietic cells and were 5-fold more resistant to BCNU and TMZ than were untransduced cells. Unirradiated hMSC present as support cells were simultaneously transduced and expressed AGT in 26% of the cells. Thus, the homogeneous nature of hMSC, and their ability to support gene transfer and be transduced themselves suggest they may be useful in clinical gene transfer protocols and have broad therapeutic applications. PMID- 10791903 TI - The selective expansion of functional T cell subsets. AB - Adoptive cellular immunotherapy is considered a potential treatment for a wide range of diseases, including cancer, infectious diseases, and autoimmune disorders. We have developed a method using a conditioned medium, XLCM, that selectively expands several different T cell subsets with a view to their use in cell therapy. Primary FBS-free suspension cultures of human peripheral blood low density mononuclear cells treated with XLCM reproducibly expand over 100,000-fold within a period of 4 weeks. CD4+ T cells and CD8+ T cells expand sequentially in the unfractionated cultures, and relatively pure populations of CD4+ and CD8+ T cells may be expanded from populations first enriched in the respective T cell subset. CD4+ T cells cultured in XLCM produce cytokines consistent with the expansion of Th1, Th2, and Th0 subsets, whereas CD8+ T cells cultured in XLCM are cytolytically competent. An interesting feature of T cells cultured in XLCM is the persistence of 5%-10% CD4+CD8+ double-positive T cells in spite of substantial single-positive T cell expansion, suggesting that these cells also proliferate in XLCM. In addition to subsets of TCRalphabeta+ T cells, TCRgammadelta+ T cells are also significantly expanded by XLCM. These results demonstrate that XLCM efficiently expands several functional T cell subsets and provides a means of obtaining selected populations suitable for use in cellular immunotherapy. PMID- 10791904 TI - Expression of the adhesion molecules CD49d and CD49e on G-CSF-mobilized CD34+ cells of patients with solid tumors or non-Hodgkin's and Hodgkin's lymphoma and of healthy donors is inversely correlated with the amount of mobilized CD34+ cells. AB - The yield of CD34+ PBPC and colony-forming units-granulocyte-macrophage (CFU-GM) in leukapheresis products and the expression of the adhesion molecules CD11a, CD31, CD49d, CD49e, CD54, CD58, CD62L, c-kit (CD117), Thy-1 (CD90), CD33, CD38, and HLA-DR on CD34+ PBPC were analyzed in patients with cancer of the testis (n = 10), breast cancer (n = 10), Hodgkin's disease (n = 20), high-grade (n = 20) and low-grade (n = 20) non-Hodgkin's lymphoma, and healthy donors (n = 20) undergoing G-CSF (filgrastim)-stimulated PBPC mobilization. For each disease entity, G-CSF was administered in two different doses, 10 microg G-CSF/kg body weight (BW)/day s.c. vs. 24 microg G-CSF/kg BW s.c./day in steady-state condition. Data were compared for each dose group separately. Patients with cancer of the testis and breast cancer mobilized significantly more CD34+ cells than patients with high grade and low-grade non-Hodgkin's lymphoma and Hodgkin's disease (p<0.05). Correspondingly, expression of CD49d on CD34+ PBPC was significantly lower in the same patients with cancer of the testis compared with high-grade and low-grade non-Hodgkin's lymphoma and Hodgkins' disease and in patients with breast cancer compared with high-grade and low-grade non-Hodgkin's lymphoma, Hodgkins's disease, and healthy donors. Similar results were obtained for CD49e. These data suggest that the expression of the adhesion molecules CD49d and CD49e on G-CSF mobilized CD34+ cells of patients with solid tumors, non-Hodgkin's lymphoma, Hodgkin's disease, and healthy donors is inversely correlated with the amount of mobilized CD34+ cells. PMID- 10791905 TI - Comparative evaluation of commonly used clones and fluorochrome conjugates of monoclonal antibodies for CD34 antigen detection. AB - CD34+ cell enumeration is currently the most appropriate technique for hematopoietic graft quality control. At the same time, numerous CD34 mAb have become commercially available. This study was designed to compare the commonly used clones 8G12 and QBEND-10 with the new clones 581 and BIRMA-K3. All available fluorochrome conjugates were tested: FITC, PE, and PE-Cy5 or PerCP for QBEND, BIRMA, 581, and 8G12 and FITC and PE for 581. Bone marrow from healthy donors (n = 5) and leukapheresis samples (n = 16) were stained, according to each manufacturer's protocol and analyzed using the FACScan. The following parameters were evaluated: % CD34+ cells detected and percentage of deviation from the median within each sample; mean channel fluorescence intensity of the CD34+ cells; resolution index (median channel fluorescence intensity of CD34+ cells/monocytes), % overlapping of CD34+ cell and monocyte fluorescence; proportion of CD34+ events after blocking with the same unlabeled clone; values of compensation requirements. Tables with results for each evaluated parameter separately were created, and rank points were applied. These scores represented the quality performance of the studied clones and fluorochrome conjugates and may be summarized as follows: 581 and 8G12 produced the best results, followed by BIRMA-K3 and QBEND10. The fluorochrome sequence was PE, PE-Cy5, PerCP, and FITC. However, all PE conjugates of the studied clones provided highly comparable results and conditions for CD34+ cell enumeration. When antigen coexpression must be studied and another dye than PE must be applied for CD34+ cell discrimination, the PE-Cy5 conjugates should be preferred. PMID- 10791907 TI - Hematotherapy literatureWatch. PMID- 10791906 TI - Two-day collection and pooling of peripheral blood stem cells with semiautomated density gradient cell separation. AB - Autologous and allogeneic PBSC collection and cryopreservation have been shown to be feasible in the pediatric population. This technique may be associated with complications, including volume overload and DMSO toxicity. To decrease these risks, we developed a technique by which PBSC are collected over a 2-day period and pooled prior to cryopreservation. PBSC are harvested on day 1 and stored with tissue culture medium on a rocker at ambient temperature. On day 2, a second PBSC harvest is performed, and the two harvests are pooled, separated on a Ficoll gradient in a semiautomatic closed system, and frozen with 10% DMSO after a soft spin for volume reduction. Cells from each day's harvest are tested for cell count and viability. A total of 36 collections in 26 patients were performed. This technique resulted in a 73%+/-0.0% reduction in volume (mean +/- SEM) and an 88%+/-0.9% depletion of RBC. Mononuclear cell (MNC) count recovery was 88%+/ 2.6%, and the MNC dose delivered to the patient was 3.1+/-0.6x10(8) cells/kg. Cell viability was >98% before and after processing. Seventeen patients have been transplanted thus far, and all these patients engrafted with minimal toxicity. These data indicate that storing PBSC for up to 24 h after harvest does not decrease PBSC viability or delay engraftment. PMID- 10791908 TI - Fifty years ago: the First International Congress of Biochemistry. PMID- 10791909 TI - The p41 fragment story. AB - The discovery of a fragment from the p41 form of invariant chain tightly bound to cathepsin L provided the first direct link between MHC class II molecules and the regulation of activity of lysosomal cysteine proteases. We recently determined the crystal structure of this p41 invariant chain fragment in complex with cathepsin L [EMBO J. 18, 793-803 (1999)]. This structure explains the specificity of the observed interactions and actually provides a tool, which can be utilized by means of molecular biology, to explore and understand the specificity of thyroglobulin type I domains and thus allow the design of specific inhibitors of papain-like cysteine proteases. The structure further supports the hypothesis that the thyroglobulin type I and II domains present in various proteins, sometimes in multiple repeats, are regulatory elements of the processing of these proteins by proteolytic cleavage. PMID- 10791910 TI - Macromolecular mimicry of nucleic acid and protein. AB - Although proteins and nucleic acids consist of different chemical components, proteins can mimic structures and possibly also functions of nucleic acids. Recently, structural mimicry was observed between two elongation factors in bacterial protein biosynthesis leading to the introduction of the concept of macromolecular mimicry. Macromolecular mimicry has further been proposed among initiation and release factors, thereby adding a new element to the description of protein synthesis in bacteria. Such mimicry has also been observed in other biological processes such as autoimmunity, DNA repair, and gene regulation, at both transcriptional and translational levels. PMID- 10791911 TI - Proteomic strategies in bladder cancer. AB - Here we review our current strategy for identifying phenotypic changes in the urothelium of bladder cancer patients with invasive disease, using a combination of proteomic technologies and immunowalking. This approach, which in principle can be applied to the study of any type of bladder cancer, makes use of proteomic technologies (for more information see http://biobase.dk/cgi-bin/celis) to reveal and identify proteins that are differentially expressed in fresh tumors and normal urothelium. Thereafter, specific antibodies against the differentially expressed proteins are used (immunowalking) to stain serial cryostat sections of cystectomies obtained from tumor-bearing patients who have undergone removal of the bladder for invasive disease. Since bladder cancer is a field disease--that is, a large part of the bladder lining is involved-the urothelium of these patients is expected to exhibit a spectrum of abnormalities, ranging from early stages of transformation to invasive disease. Besides highlighting the problems one faces when applying powerful proteomic technologies to the study of heterogeneous biopsy material, these studies show that it is feasible to study bladder cancer under experimental conditions that closely resemble the in vivo situation. PMID- 10791912 TI - S-nitrosylation of viral proteins: molecular bases for antiviral effect of nitric oxide. AB - Nitric oxide (NO) is considered an important signaling molecule implied in various different physiological processes, including nervous transmission, vascular regulation, and immune defence, as well as the pathogenesis of several diseases. NO reportedly also has an antiviral effect on several DNA and RNA virus families. The NO-mediated S-nitrosylation of viral and host (macro)molecules appears to be an intriguing general mechanism for the control of the virus life cycle. In this respect, NO is able to nitrosylate cysteine-containing enzymes (e.g., proteases, reverse transcriptase, and ribonucleotide reductase). Moreover, zinc-fingers and related domains present in enzymes (e.g., HIV-1-encoded integrase or herpes simplex virus type-1 heterotrimeric helicase-primase complex) or nucleocapsid proteins may be considered as NO targets. Also, NO may regulate both host (e.g., nuclear factor-kappaB) and viral-encoded (e.g., HIV-1 tat protein or Epstein-Barr virus Zta) transcriptional factors that are involved in virus replication. Finally, NO-mediated S-nitrosylation of cysteine-containing glycoproteins and hemagglutinin may also occur. Here, NO targets are summarised, and the molecular bases for the antiviral effect of NO are discussed. PMID- 10791913 TI - Presenilins: multifunctional proteins involved in Alzheimer's disease pathology. AB - Early-onset aggressive forms of Alzheimer's disease (AD) are of genetic nature and have been linked to inherited mutations located on chromosomes 21, 14, and 1. The gene products of chromosomes 14 and 1, which are responsible for most of these familial forms of the disease (FAD), have been recently identified and referred to as presenilin 1 and 2 (PS1, PS2), respectively. Several lines of evidence derived from neuropathological, cell biology, and transgenesis approaches indicate that PS could interfere with the processing of the beta amyloid precursor protein (betaAPP). Thus, FAD-linked mutations in PS exacerbate the production of Abeta42, the readily aggregable Abeta species corresponding to one of the main constituents of the senile plaques that invade the cortical areas of affected brains. Recent studies indicate that PS functions could be intimately related with the susceptibility of PS to further processing by caspase-like enzymes and other unknown proteolytic activities. Here I briefly report on the post-translational events undergone by PS and examine recent advances concerning their possible roles in development, cell signaling, and apoptosis. Possible alterations brought by FAD-linked mutations will be discussed. PMID- 10791914 TI - The broad spectrum of responses to oxidants in proliferating cells: a new paradigm for oxidative stress. AB - Proliferating mammalian cells exhibit a broad spectrum of responses to oxidative stress, depending on the stress level encountered. Very low levels of hydrogen peroxide, e.g., 3 to 15 microM, or 0.1 to 0.5 micromol/10(7) cells, cause a significant mitogenic response, 25% to 45 % growth stimulation. Greater concentrations of H2O2, 120 to 150 microM, or 2 to 5 micromol/10(7) cells, cause a temporary growth arrest that appears to protect cells from excess energy use and DNA damage. After 4-6 h of temporary growth arrest, many cells will exhibit up to a 40-fold transient adaptive response in which genes for oxidant protection and damage repair are preferentially expressed. After 18 h of H2O2 adaptation (including the 4-6 h of temporary growth arrest) cells exhibit maximal protection against oxidative stress. The H2O2 originally added is metabolized within 30-40 min, and if no more is added the cells will gradually de-adapt, so that by 36 h after the initial H2O2 stimulus they have returned to their original level of H2O2 sensitivity. At H2O2 concentrations of 250 to 400 microM, or 9 to 14 micromol/10(7) cells, mammalian fibroblasts are not able to adapt but instead enter a permanently growth-arrested state in which they appear to perform most normal cell functions but never divide again. This state of permanent growth arrest has often been confused with cell death in toxicity studies relying solely on cell proliferation assays as measures of viability. If the oxidative stress level is further increased to 0.5 to 1.0 mM H2O2, or 15 to 30 micromol/10(7) cells, apoptosis results. This oxidative stress-induced apoptosis involves nuclear condensation, loss of mitochondrial transmembrane potential, degradation/down-regulation of mitochondrial mRNAs and rRNAs, and degradation/laddering of both nuclear and mitochondrial DNA. At very high H2O2 concentrations of 5.0 to 10.0 mM, or 150 to 300 micromol/10(7) cells and above, cell membranes disintegrate, proteins and nucleic acids denature, and necrosis swiftly follows. Cultured cells grown in 20% oxygen are essentially preadapted or preselected to survive under conditions of oxidative stress. If cells are instead grown in 3% oxygen, much closer to physiological cellular levels, they are more sensitive to an oxidative challenge but exhibit far less accumulated oxidant damage. This broad spectrum of cellular responses to oxidant stress, depending on the amount of oxidant applied and the concentration of oxygen in the cell culture system, provides for a new paradigm of cellular oxidative stress responses. PMID- 10791915 TI - Identification of a novel cytosolic tocopherol-binding protein: structure, specificity, and tissue distribution. AB - Alpha-tocopherol plays an important role as a lipid-soluble antioxidant. It is present in all major mammalian cell types and shows tissue-specific distribution. This suggests the presence of specific proteins involved in intracellular distribution or metabolism of alpha-tocopherol. A diminution of tocopherol plasma concentrations contributes to the development of diseases such as vitamin E deficiency (AVED), atherosclerosis, and prostate cancer. Further evidence has been obtained for the existence of sites in cellular metabolism and signal transduction where alpha-tocopherol potentially plays a regulatory role. A signal transduction modulation specific for alpha-tocopherol has been described in several model systems. Using radioactively labeled alpha-tocopherol as tracer, we have isolated a new alpha-tocopherol-associated protein (TAP) from bovine liver. This protein has a molecular mass of 46 kDa and an isoelectric point of 8.1. From its partial amino acid sequence, a human gene has been identified with high homology to the newly described protein. Sequence analysis has established that the new TAP has structural motifs suggesting its belonging to a family of hydrophobic ligand-binding proteins (RALBP, CRALBP, alpha-TTP, SEC 14, PTN 9, RSEC 45). Human TAP has been cloned into Escherichia coli, and its tissue specific expression has been assessed by Northern blot analysis. PMID- 10791916 TI - Redox cycles of caffeic acid, alpha-tocopherol, and ascorbate: implications for protection of low-density lipoproteins against oxidation. AB - This study addresses the dynamic interactions among alpha-tocopherol, caffeic acid, and ascorbate in terms of a sequence of redox cycles aimed at accomplishing optimal synergistic antioxidant protection. Several experimental models were designed to examine these interactions: UV irradiation of alpha-tocopherol containing sodium dodecyl sulfate micelles, one-electron oxidations catalyzed by the hypervalent state of myoglobin, ferrylmyoglobin, and autoxidation at appropriate pHs. These models were assessed by ultraviolet (UV) and electron paramagnetic resonance (EPR), entailing direct- and continuous-flow experiments, spectroscopy and by separation and identification of products by HPLC. The alpha tocopheroxyl radical EPR signal generated by UV irradiation of alpha-tocopherol containing micelles was suppressed by caffeic acid and ascorbate; in the former case, no other EPR signal was observed at pH 7.4, whereas in the latter case, the alpha-tocopheroxyl radical EPR signal was replaced by a doublet EPR spectrum corresponding to the ascorbyl radical (A*-). The potential interactions between caffeic acid and ascorbate were further analyzed by assessing, on the one hand, the ability of ascorbate to reduce the caffeic acid o-semiquinone (generated by oxidation of caffeic acid by ferrylmyoglobin) and, on the other hand, the ability of caffeic acid to reduce ascorbyl radical (generated by autoxidation or oxidation of ascorbate by ferrylmyoglobin). The data presented indicate that the reductive decay of ascorbyl radical (A*-) and caffeic acid o-semiquinone (Caf-O*) can be accomplished by caffeic acid (Caf-OH) and ascorbate (AH-), respectively, thus pointing to the reversibility of the reaction Caf-O* + AH- <--> Caf-OH + A* . Continuous-flow EPR measurements of mixtures containing ferrylmyoglobin, alpha tocopherol-containing micelles, caffeic acid, and ascorbate revealed that ascorbate is the ultimate electron donor in the sequence encompassing transfer of the radical character from the micellar phase to the phase. In independent experiments, the effects of caffeic acid and ascorbate on the oxidation of two low-density lipoprotein (LDL) populations, control and alpha-tocopherol-enriched, were studied and results indicated that alpha-tocopherol, caffeic acid, and ascorbate acted synergistically to afford optimal protection of LDL against oxidation. These results are analyzed for each individual antioxidant in terms of three domains: its localization and that of the antioxidant-derived radical, its reduction potential, and the predominant decay pathways for the antioxidant derived radical, that exert kinetic control on the process. PMID- 10791917 TI - Expression and characterization of chicken muscle ecto-ATPase in mammalian COS cells. AB - Chicken muscle ecto-ATPase has unusual enzyme kinetics and properties not found in many other E-type ATPases. To determine whether the unique properties of the chicken ecto-ATPase are inherent in the protein sequence and not mediated by some unique property of the chicken system, we have spliced together two partial cDNAs encoding the ecto-ATPase. The enzymatic properties of the COS (green monkey kidney) cell-expressed protein are indistinguishable from the purified chicken gizzard ecto-ATPase, including a 2- to 3-fold stimulation of membrane-bound activity by crosslinking and lectins, properties not shared by most other E-type ATPases. The expressed enzyme is specific for nucleotide triphosphates (ATPase:ADPase hydrolysis ratio of 26:1) and is inhibited by Cibacron Blue (IC50 = 10 microM). The active, expressed enzyme can be affinity-purified with Cibacron Blue, is relatively resistant to deglycosylation, and is less stable than other E type ATPases. Expression in the presence of tunicamycin resulted in an inactive, unfolded enzyme. PMID- 10791918 TI - Fibroblast growth factor receptor-4 splice variants cause deletion of a critical tyrosine. AB - We have identified two novel isoforms of fibroblast growth factor receptor-4 (FGFR4). They result from alternative splicing of intron 17. Two transcripts, both slightly larger than the one coding for the known mouse FGFR4, are generated. The shortest (FGFR4-17a) includes the 31-most 3'-nucleotides of intron 17; the longest (FGFR4-17b) includes all 114 nucleotides of intron 17. Translation of the FGFR4-17a and FGFR4-17b splice variants predicts that both novel putative FGFR4 isoforms have a truncated C-terminal intracellular tail. The first amino acid residue affected by the insertions in both novel isoforms is Tyr 760, a residue that may play a crucial role in intracellular signaling through stimulation of the phosphatidylinositol-biphosphate pathway. PMID- 10791919 TI - Involvement of histone phosphorylation in thymocyte apoptosis by protein phosphatase inhibitors. AB - Incubation of rat thymocytes with the inhibitors of protein phosphatase such as calyculin A and okadaic acid resulted in an increase in DNA fragmentation. These effects were dependent on the concentration of the inhibitors and the incubation time. Analyses of the fragmented DNA revealed the production of approximately 50 kbp of DNA and a 180 bp DNA ladder. In addition, a laser scanning-microscopic analysis showed that these compounds caused nuclear condensation. Thus, these results demonstrated that protein phosphatase inhibitors induced thymocyte apoptosis. The inhibitors of protein phosphatase increased the phosphorylation of proteins of approximately 15 kDa. The phosphorylation of proteins preceded the DNA fragmentation induced by these inhibitors. Judging from acetic acid-urea Triton X-100 gel electrophoresis, the phosphorylated proteins were histone H1 and H2A/H3. Therefore, these results suggest that phosphorylation of histones triggers the DNA fragmentation of thymocytes undergoing apoptosis. PMID- 10791920 TI - pp60v-src reactivation inhibits serum-induced accumulation of inositol phosphates and phosphatidylethanol in tsNRK. AB - In tsRSV-infected NRK (tsNRK) cells, pp60(v-src) reactivation by temperature shift from a nonpermissive temperature, 39 C, to a permissive one, 32 degrees C, induced the production of inositol phosphates (IPt) and phosphatidylethanol (PEt). This was accompanied by an increase in membrane-associated protein kinase C (PKC) activity in the absence of exogenous growth factors. However, with serum stimulation, the amounts of IPt and PEt at 32 degrees C were less than those at 39 degrees C. Pretreatment with PKC inhibitors, Ro-31-8220 and staurosporine, enhanced the accumulation of IPt but not of PEt at 32 degrees C. The tyrosine phosphorylation of phospholipase Cgamma1 (PLCgamma1) was increased either by serum or by pp60(v-src) reactivation. These results suggest that serum transduces its signal through PLCgamma1 mediation, and that pp60(v-src), possibly through the PKC mediation, negatively affects serum-induced PLCgamma1 activation. PMID- 10791921 TI - A model of EcoRII restriction endonuclease action: the active complex is most likely formed by one protein subunit and one DNA recognition site. AB - To elucidate the mechanism of interaction of restriction endonuclease EcoRII with DNA, we studied by native gel electrophoresis the binding of this endonuclease to a set of synthetic DNA-duplexes containing the modified or canonical recognition sequence 5'-d(CCA/TGG)-3'. All binding substrate or substrate analogues tested could be divided into two major groups: (i) duplexes that, at the interaction with endonuclease EcoRII, form two types of stable complexes on native gel in the absence of Mg2+ cofactor; (ii) duplexes that form only one type of complex, observed both in the presence and absence of Mg2+. Unlike the latter, duplexes under the first group can be hydrolyzed by endonuclease. Data obtained suggest that the active complex is most likely formed by one protein subunit and one DNA recognition sequence. A model of EcoRII endonuclease action is presented. PMID- 10791922 TI - cDNA cloning of the two subunits of phospholipase A2 inhibitor PLIgamma from blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus. AB - Three phospholipase A2 (PLA2) inhibitors (PLI) have been purified from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus; 1 of these, PLIgamma, contains 2 homologous subunits, PLIgamma-A and PLIgamma-B. The cDNAs encoding these 2 subunits of PLIgamma were isolated from a liver cDNA library by using fragments from polymerase chain reaction amplifications as probes and sequenced. The respective nucleotide sequences encoded 19-residue signal sequences, followed by 181-residue proteins. The calculated molecular masses were 20123 and 20150 Da for the PLIgamma-A and PLIgamma-B subunits, respectively; and PLIgamma-A included a N-linked carbohydrate site at Asn-157. The sequences of these subunits contained 2 internal repeats of disulfide-bonding pattern characteristic to those of urokinase-type plasminogen activator receptor and members of the Ly-6 superfamily. A phylogenetic analysis comparing the amino acid sequences of PLIgamma-A and PLIgamma-B with those for other snakes revealed that the gene duplication leading to these 2 subunits occurred before the divergence of Viperidae and Elapidae. PMID- 10791923 TI - Sphingomyelinase activity of livers from control and NCTR-BALB/c mice. AB - NCTR-BALB/c mice have an autosomal recessive disorder involving storage of sphingomyelin and unesterified cholesterol in their tissues and reduced tissue sphingomyelinase activity. With [N-methyl-14C]sphingomyelin as substrate, Vmax for the enzyme in livers from control and mutant mice were, respectively, 29.6 and 11.6 nmol of substrate hydrolyzed h(-1) mg(-1) protein, and the corresponding Km values were 94.6 and 132.3 microM. The control and mutant enzymes showed similar pH profiles and temperature sensitivities. When the control and mutant liver homogenates were mixed in various proportions, the resulting activities were 70-80% of the theoretical values. Cross and straight addition of total lipid extracts of control and mutant livers had minimal effect on their enzyme activities. The results suggest that the reduced sphingomyelinase activity of mutant liver is not due to the presence of inhibitors or absence of activators in this tissue. PMID- 10791924 TI - Effects of genetic and diet-induced obesity on lipid metabolism. AB - C57BL/6J obese (ob/ob) and lean mice fed ad libitum on a normal mouse chow diet (Normal), were compared with lean mice of the same age and strain fed ad libitum on a high-fat diet, consisting of the Normal diet with the addition of beef lard (Lard), from age 3 months for 34 days. The lard-fed mice were seen to have significantly higher (P<0.05) body weight in this 34-day period than that of the other two groups fed on the Normal diet. Epididymal fat depot and adipocyte cell size were significantly larger (P<0.05) in the Lard-fed lean mice and in the obese (ob/ob) mice than were those of the Normal-fed lean mice. Dietary Lard intake did not significantly affect concentrations of plasma triglyceride although those of plasma cholesterol were significantly increased (P<0.05). The development of obesity in these Lard-fed mice appeared to be accelerated and significant. PMID- 10791925 TI - Expression of p21WAF1/CIP1 in bladder cancer: relation to schistosomiasis. AB - Cell cycle regulation is mediated in part through expression of the cyclin dependent kinase inhibitor p21WAF1/CIP1. Loss of p21WAF1/CIP1 expression may, therefore, contribute partially to schistosomal carcinogenesis in the urinary bladder. We compared p21WAF1/CIP1 expression in schistosomal and nonschistosomal bladder cancer to explore possible differences in p21WAF1/CIP1 expression between the two subtypes and the possible association between schistosomiasis and loss of p21WAF1/CIP1 expression. Tumor specimens were obtained from 130 patients who underwent transurethral biopsy or cystectomy. p21WAF1/CIP1 was determined by immunodot blot, Western blot, and enzyme immunoassay (EIA). We validated a highly sensitive quantitative EIA assay for determination of p21WAF1/CIP1 in cell lysates. Precision, analytical recovery, and linearity were all excellent. Our results did not show any correlation between p21WAF1/CIP1 expression and most clinicopathologic variables. Lower expression of p21WAF1/CIP1 was evident in squamous cell carcinoma (SCC) and schistosomal subtype than in transitional cell carcinoma and nonschistosomal tumors. Our data suggest a potential role for p21WAF1/CIP1 alteration in schistosomal carcinogenesis. PMID- 10791926 TI - Suppression of caspase-3 and nitric-oxide synthase-2 during buccal mucosal ulcer healing: effect of chronic alcohol ingestion. AB - In this study, we analyzed the effect of chronic alcohol ingestion on the expression of constitutive (cNOS) and inducible (NOS-2) nitric-oxide synthase and the activity of an apoptotic protease, caspase-3, during buccal mucosal ulcer healing in rats maintained for 5 weeks on alcohol-containing or control liquid diet. In comparison with the controls, the ulcer onset in the alcohol group was characterized by a 2.5-fold greater epithelial cells apoptosis, 2.1-fold greater expression of caspase-3 activity, and a 1.4-fold greater enhancement in NOS-2, but expression of cNOS showed a 1.3-fold decrease. In both groups the ulcer healing was accompanied by a gradual decline in apoptosis, caspase-3, and NOS-2 and a recovery in cNOS activity, but the changes were considerably slower in the alcohol diet group, as manifested by a 40%(4 days) delay in ulcer healing. These results suggest that chronic alcohol ingestion interferes with the suppression of NOS-2 and the apoptotic events propagated by caspase-3 and hence affects the efficiency of oral mucosal repair process. PMID- 10791927 TI - Detection of specific protein bands with melatonin-like immunoreactivity in different cell lines and human brain regions. AB - The pineal hormone melatonin regulates various neural and endocrine processes involved in mammalian circadian rhythms. To understand how melatonin mediates these functions, we investigated melatonin-like immunoreactivity (MLI) in cell extracts and human brain. In Western immunoblots, we detected high-molecular-mass protein bands (85-135 kDa) that specifically reacted with the anti-melatonin antibody. The specific protein bands were present in cell extracts from the human brain and cell lines of different origins. The immunoreactive signal of the 135 kDa protein band was highest in a neuroendocrine PC12 cell line, which was 10 fold higher than the signal observed in any cell extracts used. The commercial antibody employed in the Western blots was further purified against serum proteins and thyroglobulins. We have previously reported that the antibody against melatonin recognizes MLI as detected by a sensitive RIA. In the present report we have detected the putative melatonin-specific binding proteins, which could contribute to the MLI. Our results suggest that melatonin binds with specific proteins in different cellular and brain extracts, the protein(s) being maximally synthesized in PC12 cells. These results may indicate a group of yet unknown proteins sharing a melatonin-like epitope or the presence of melatonin binding protein(s) that regulate availability of free melatonin, or both. PMID- 10791928 TI - Comparison of clinical signs and hemodynamic variables used to monitor rabbits during halothane- and isoflurane-induced anesthesia. AB - OBJECTIVE: To characterize variables used to monitor rabbits during inhalation anesthesia. ANIMALS: 8 male New Zealand White rabbits. PROCEDURE: Rabbits were similarly anesthetized with halothane (HAL) or isoflurane (ISO) in a crossover study; half received HAL followed by ISO, and the protocol was reversed for the remaining rabbits. After induction, minimum alveolar concentration (MAC) was determined for each agent, using the tail-clamp method, and variables were recorded at 0.8, 1.0, 1.5, and 2.0 MAC (order randomized). RESULTS: Mean +/- SEM MAC was 1.42 +/- 0.05 and 2.07 +/- 0.09% for HAL and ISO, respectively. Directly measured auricular mean arterial blood pressure was 52.8 +/- 5.6 and 54.8 +/- 6.1 mm Hg at 0.8 MAC for HAL and ISO, respectively, and decreased from these values in a parallel dose-dependent manner. Respiratory frequency remained constant (range, 69 to 78 breaths/min) over the range of HAL doses but incrementally decreased from a mean of 53 (at 0.8 MAC) to 32 breaths/min (at 2.0 MAC) for ISO. The PaCO2 was similar at 0.8 MAC for HAL and ISO and progressively increased with increasing doses of both agents; PaCO2 at 2.0 MAC for ISO was significantly greater than that at 2.0 MAC for HAL (79.8 +/- 13.7 vs 54.9 +/- 4.0 mm Hg, respectively). Eyelid aperture consistently increased in a dose-dependent manner for both anesthetics. CONCLUSIONS: Arterial blood pressure, PaCO2, and eyelid aperture consistently and predictably changed in rabbits in response to changes in anesthetic doses. The magnitude of respiratory depression was greater for ISO than for HAL. PMID- 10791929 TI - Factors associated with time until first race and career duration for Thoroughbred racehorses. AB - OBJECTIVE: To determine whether sex or foaling period were associated with time to first race start and whether sex, age at time of first race start, or inter race period were associated with career duration for Thoroughbred racehorses in Australia. ANIMALS: 553 Thoroughbred racehorses. PROCEDURE: Race records through to the end of the 6-year-old racing season were obtained for each horse. Product limit survival analysis was used to determine whether sex or foaling period was associated with time to first race start and whether sex or age at first race was associated with career duration. A Cox proportional hazards model was used to determine whether sex, age at first possible race, age at first race, or inter race period was associated with time to first race start or career duration. RESULTS: 472 horses (85.4%) raced. Time to first race start was not associated with sex or foaling period. However, males, who had their first race at a younger age and a greater mean number of rest days between races, had longer careers. The hazard ratio for career duration of males versus females, controlling for age at time of first race and inter-race period (0.27), confirmed that males had a higher probability of a long racing career. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for Thoroughbred racehorses, sex and time of birth during the foaling season were not associated with time to first race. Male Thoroughbred racehorses that were raced less intensively or raced at a younger age were more likely to have long racing careers. PMID- 10791930 TI - Effects of the antimicrobial growth promoter tylosin on subclinical infection of pigs with Salmonella enterica serotype Typhimurium. AB - OBJECTIVE: To determine whether feeding tylosin, an antimicrobial growth promoter, to pigs was associated with increased risk of infection with and excretion of Salmonella enterica serotype Typhimurium. ANIMALS: 17 healthy pigs. PROCEDURE: A commercial pelleted dry feed was given in 2 feeding trials. In trial A, 11 pigs were given feed with tylosin, 11 pigs were given feed without tylosin, and 11 pigs were given feed with tylosin before and feed without tylosin after inoculation with S Typhimurium. In trial B, 44 pigs were given feed that contained tylosin, and 44 pigs were given feed without tylosin. Three weeks after the start of each trial, pigs were orally inoculated with approximately 5 x 10(6) colony-forming units of S Typhimurium. Feces were examined for S Typhimurium, using semiquantitative microbiologic techniques before and for 5 or 6 weeks after inoculation. Serum antibody titers against S enterica were measured by use of ELISA. RESULTS: None of the pigs developed clinical signs of salmonellosis. However, after inoculation, S Typhimurium was isolated from feces of most pigs, and all but 2 pigs developed serum antibodies against S enterica. Significant differences were not detected between experimental and control groups in either trial. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that tylosin fed as an antimicrobial growth promoter to pigs may not be an important factor in promoting infection with or excretion of S enterica serotype Typhimurium. PMID- 10791931 TI - Effect of inhaled nitric oxide on experimentally induced pulmonary hypertension in neonatal foals. AB - OBJECTIVE: To evaluate the efficacy of inhaled nitric oxide (NO) in anesthetized healthy newborn foals with experimentally induced pulmonary hypertension. ANIMALS: Five 1- to 3-day-old foals. PROCEDURE: Anesthesia was induced and maintained with propofol, and foals were intubated and mechanically ventilated. Systemic pressure and pulmonary arterial pressure (P(PA)) were recorded every 30 seconds. Hypertension was induced via a hypoxic gas mixture or chemical vasoconstriction, using the thromboxane mimetic U46619. Nitric oxide was added at a concentration of 80 parts per million (ppm) for 6 minutes under baseline conditions and during pulmonary hypertension-induced alveolar hypoxia (inspired oxygen concentration = 0.08). Nitric oxide (20, 40, 80, and 160 ppm) was evaluated during U46619-induced hypertension. Samples for determination of arterial blood gas tensions were collected before and after each NO treatment. RESULTS: Inhaled NO (approx 80 ppm) did not have an effect on baseline variables. Infusion of U46619 (0.35 +/- 0.04 microg/kg of body weight/min) or alveolar hypoxia resulted in increased P(PA) and decreased arterial oxygenation (PaO2) and hemoglobin saturation (HbSat). The increase in P(PA) was attenuated, in a dose dependent manner, by NO during U46619 infusion and reversed by NO during induced hypoxemia. The PaO2 and HbSat were significantly improved at all NO doses during U44619 infusion but not during alveolar hypoxia. For all inhaled NO concentrations, nitrogen dioxide and methoglobin values were < 5 ppm and 3%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Nitric oxide is a potent, selective vasodilator of the pulmonary circulation in healthy newborn foals. Inhaled NO may have value as a therapeutic agent in foals with pulmonary hypertension. PMID- 10791932 TI - A technique for evaluating analgesia in conscious perching birds. AB - OBJECTIVE: To develop a technique for objective assessment of modulation of nociperception in conscious perching birds. ANIMALS: 31 adult African grey parrots. PROCEDURE: Birds were randomly assigned to receive saline (0.9% NaCl) solution (n = 10), butorphanol tartrate (11), or buprenorphine hydrochloride (10), i.m. Birds were fitted with a surface electrode on the medial metatarsus of 1 leg. An electrical stimulus was delivered to the bird's foot through an aluminum surface on half of the perch. The alternate side of the perch delivered a noxious thermal stimulus. A withdrawal response to either stimulus was recorded when the bird lifted its foot or vigorously flinched its wings. RESULTS: Responses to thermal stimuli were extremely variable during baseline testing and after administration of drugs. Thus, significant differences were not detected after drug injection. In contrast, responses to an electrical stimulus were predictable with much less variation. CONCLUSIONS AND CLINICAL RELEVANCE: This method and device allowed for the reliable determination of withdrawal threshold in perching birds. Use of this technique for objective assessment of modulation of nociperception in conscious perching birds will enable assessment of analgesic drugs. PMID- 10791933 TI - Analgesic effects of butorphanol and buprenorphine in conscious African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh). AB - OBJECTIVE: To evaluate effects of butorphanol tartrate and buprenorphine hydrochloride on withdrawal threshold to a noxious stimulus in conscious African grey parrots. ANIMALS: 29 African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh). PROCEDURE: Birds were fitted with an electrode on the medial metatarsal region of the right leg, placed into a test box, and allowed to acclimate. An electrical stimulus (range, 0.0 to 1.46 mA) was delivered to each bird's foot through an aluminum perch. A withdrawal response was recorded when the bird lifted its foot from the perch or vigorously flinched its wings. Baseline threshold to a noxious electrical stimulus was determined. Birds then were randomly assigned to receive an i.m. injection of saline (0.9% NaCl) solution, butorphanol (1.0 mg/kg of body weight), or buprenorphine (0.1 mg/kg), and threshold values were determined again. RESULTS: Butorphanol significantly increased threshold value, but saline solution or buprenorphine did not significantly change threshold values. CONCLUSIONS AND CLINICAL RELEVANCE: Butorphanol had an analgesic effect, significantly increasing the threshold to electrical stimuli in African grey parrots. Buprenorphine at the dosage used did not change the threshold to electrical stimulus. Butorphanol provided an analgesic response in half of the birds tested. Butorphanol would be expected to provide analgesia to African grey parrots in a clinical setting. PMID- 10791934 TI - Determination of solid- and liquid-phase gastric emptying half times in cats by use of nuclear scintigraphy. AB - OBJECTIVE: To use nuclear scintigraphy to establish a range of gastric emptying half times (t1/2) following a liquid or solid meal in nonsedated cats. ANIMALS: 12 clinically normal 3-year-old domestic shorthair cats. PROCEDURE: A test meal of 75 g of scrambled eggs labeled with technetium Tc 99m tin colloid was fed to 10 of the cats, and solid-phase gastric emptying t1/2 were determined by use of nuclear scintigraphy. In a separate experiment, 8 of these cats plus an additional 2 cats were fed 18 ml (n = 5) or 36 ml (n = 5) of a nutrient liquid meal labeled with technetium Tc 99m pentetate. Liquid-phase gastric emptying t1/2 then were determined by use of scintigraphy. RESULTS: Solid-phase gastric emptying t1/2 were between 210 and 769 minutes (median, 330 minutes). Median liquid-phase gastric emptying t1/2 after ingestion of 18 or 36 ml of the test meal were 67 minutes (range, 60 to 96 minutes) and 117 minutes (range, 101 to 170 minutes), respectively. The median t1/2 determined for cats receiving 18 ml of the radiolabeled liquid was significantly less than that determined for cats receiving 36 ml of the test meal. CONCLUSIONS AND CLINICAL RELEVANCE: The protocol was tolerated by nonsedated cats. Solid-phase gastric emptying t1/2 were prolonged, compared with liquid-phase t1/2, and a major factor governing the emptying rate of liquids was the volume consumed. Nuclear scintigraphy may prove useful in assessing gastric motility disorders in cats. PMID- 10791935 TI - Isolation of bovine respiratory coronaviruses from feedlot cattle and comparison of their biological and antigenic properties with bovine enteric coronaviruses. AB - OBJECTIVE: To isolate bovine coronaviruses from the respiratory tracts of feedlot cattle and compare antigenic and biological properties of these strains with bovine enteric coronaviruses. ANIMALS: 5- to 8-month-old mixed-breed cattle at 4 feedlots. PROCEDURE: Samples were obtained from the nasal passages for testing. The 13 samples with the highest magnitude of positive values for bovine coronavirus (BCV) were cultured. Ten strains of bovine respiratory coronavirus (BRCV) were adapted successfully to serial passage. After observation of cytopathic effects (CPE) and confirmation of BRCV by immune electron microscopy and immunofluorescence testing, cell culture-adapted strains were cloned by limiting dilution. These isolates then were compared with a panel of bovine enteric coronaviruses (BECV), using hemagglutination (HA), receptor-destroying enzyme activity (RDE), hemagglutination inhibition (HI), and virus neutralization (VN) assays. Antigenic relatedness values then were calculated. RESULTS: The BRCV were detected in 105 of 488 (21.5%) of the cattle tested. Of 13 strains tested, 10 were isolated in cell culture. Six of the BRCV strains were similar to 2 strains obtained from neonatal calves with diarrhea and 2 strains from adult cattle with winter dysentery. The other 4 BRCV isolates had high RDE activity against mouse erythrocytes but differed from other strains of BECV Nine of 10 BRCV isolates had properties similar to the 2 BECV subtypes. CONCLUSIONS AND CLINICAL RELEVANCE: The BRCV can be isolated from nasal passages of cattle entering feedlots. Most BRCV were similar to BECV strains, although a few had unique properties. Vaccines developed to protect against enteric strains also may protect against respiratory tract strains. PMID- 10791936 TI - Primary nucleotide structure of predominant and alternate splice forms of equine insulin-like growth factor I and their gene expression patterns in tissues. AB - OBJECTIVE: To isolate, clone, and determine primary nucleotide sequence of equine insulin-like growth factor I (IGF-I) and to examine IGF-I gene expression in tissues and cartilage from horses. ANIMALS: Horses of various ages. METHODS: Total RNA was isolated from tissues and purified. Complementary DNA (cDNA) was derived by reverse transcription and polymerase chain reaction (PCR) amplification and subcloned to plasmid vectors for sequencing and comparison with other species. Total RNA from various tissues was probed with radiolabeled cDNA or complimentary RNA constructs by use of northern blotting, tube hybridization, or PCR procedures to determine IGF-I expression patterns. RESULTS: Nucleotide sequence of equine IGF-I was 90% homologous to that of cows, 88% homologous to that of humans and sheep, and 77% homologous to that of rats. Amino acid sequence was identical to that of humans, cows, dogs, and pigs. A larger PCR product (IGF IB) was consistent with alternate splicing with retention of IGF-I exon 4 sequence, similar to rats and mice. Northern blot analysis revealed multiple IGF I transcripts; predominant sizes were 1.6 and 4.5 kb. The IGF-I message was commonly detected in liver, kidney, and cartilage from young foals and was diminished in cartilage from a 12-month-old horse. CONCLUSIONS: Nucleotide sequences of equine prepropeptides were different from those of other species, but the sequence coding the mature IGF-I peptide was more closely homologous. The larger IGF-IB form differed substantially in the carboxy-terminal. The biological action of the cleaved terminal was speculated to be autocrine feedback. Expression of IGF-I was apparent in many tissues, including cartilage, and was greater in immature horses. PMID- 10791937 TI - Pharmacologic effects of epidural versus intramuscular administration of detomidine in cattle. AB - OBJECTIVES: To determine whether epidural administration of detomidine hydrochloride to cattle induced analgesia of the perineum and to compare analgesic and systemic effects of epidural versus i.m. administration of detomidine at a dose of 40 microg/kg in cattle. ANIMALS: 18 healthy adult cows. PROCEDURE: 6 cows were given detomidine by epidural administration, 6 were given detomidine i.m., and 6 (control group) were not given detomidine. Analgesia was assessed by determining responses to needle pinpricks in the perineum and flank and by applying electrical stimuli to the perineum and flank and determining the voltage that induced an avoidance response. Degree of sedation and ataxia were scored, and mean arterial pressure, heart rate, respiratory rate, and frequency of ruminal contractions were measured. RESULTS: Epidural and i.m. administration of detomidine induced comparable degrees of analgesia of the perineum and flank, accompanied by moderate sedation and ataxia, hypertension, cardiorespiratory depression, and rumen hypomotility. CONCLUSIONS AND CLINICAL RELEVANCE: Epidural and i.m. administration of detomidine at a dose of 40 microg/kg induced similar analgesic and systemic effects in cattle. Epidural administration of detomidine did not appear to be advantageous over i.m. administration. PMID- 10791938 TI - Comparison of three methods for estimation of exercise-related ion losses in sweat of horses. AB - OBJECTIVE: To quantify total fluid loss in sweat of Thoroughbreds during >3 hours of low-intensity exercise in controlled conditions and to calculate and compare estimated ion losses in sweat, according to 3 methods. ANIMALS: 6 exercise trained Thoroughbreds. PROCEDURE: Fluid and ion losses in sweat were measured in 6 horses exercising at 40% of the speed that elicited maximum oxygen consumption for 45 km. Horses were given a 15-minute rest period at the end of three 15-km exercise phases. Horses completed 2 exercise trials. Ion losses in sweat were calculated, using measurements of local sweating rate and sweat ion composition (SWT), change in net exchangeable cation content (CAT), and change in extracellular ion content (PLAS) derived from plasma total solids and ion concentrations. RESULTS: Measurement of SWT revealed a mean (+/- SEM) fluid loss in sweat during 45 km of exercise of 27.5 +/- 1.6 L. Total ion loss in sweat was approximately 241 g or 7.8 mol with higher sodium losses in the second and third phases of exercise compared with the first phase. Losses of sodium and potassium calculated by SWT or CAT were not significantly different from each other, whereas losses of these ions as determined by PLAS were significantly lower. CONCLUSIONS AND CLINICAL RELEVANCE: Calculation of ion losses from a mean whole body sweating rate extrapolated from either local sweating rate and sweat ion composition or from change in net exchangeable cation content provide similar results, whereas ion losses determined by changes in extracellular ion content derived from plasma total solids and ion concentration results in underestimation of actual losses. PMID- 10791939 TI - Analysis of surface antigen expression and host defense function in leukocytes from calves heterozygous or homozygous for bovine leukocyte adhesion deficiency. AB - OBJECTIVE: To analyze surface antigen expression and functional responses of leukocytes from calves heterozygous and homozygous for bovine leukocyte adhesion deficiency (BLAD). ANIMALS: 8 clinically normal calves, 4 calves heterozygous for BLAD, and 4 calves homozygous for BLAD. PROCEDURE: Surface antigen expression was examined by flow cytometric analysis of leukocytes stained with monoclonal antibodies. Neutrophil function analyses included phagocytosis and killing of Candida albicans and measurement of respiratory burst activity using cytochrome c and dihydrorhodamine 123 assays. Differential leukocyte counts also were performed. RESULTS: Leukocytes from heterozygous calves were similar to those of clinically normal calves with respect to surface antigen expression, C albicans phagocytosis and killing, and respiratory burst activity. In contrast, neutrophils from calves homozygous for BLAD had significantly reduced phagocytic and yeast-killing capacity but had higher respiratory burst activity than cells from clinically normal or heterozygous calves. Homozygous calves also had extreme neutrophilia and significantly more immature neutrophils. CONCLUSIONS: The heterozygous BLAD genotype does not cause detectable functional differences in leukocytes, compared with those of clinically normal calves. In contrast, leukocytes from homozygous calves seem to upregulate alternative host defense capabilities (eg, respiratory burst activity) to partially compensate for the lack of typical adherence-dependent host defense functions. PMID- 10791941 TI - Cardiorespiratory and metabolic effects of xylazine, detomidine, and a combination of xylazine and acepromazine administered after exercise in horses. AB - OBJECTIVE: To determine sedative, cardiorespiratory and metabolic effects of xylazine hydrochloride, detomidine hydrochloride, and a combination of xylazine and acepromazine administered i.v. at twice the standard doses in Thoroughbred horses recuperating from a brief period of maximal exercise. ANIMALS: 6 adult Thoroughbreds. PROCEDURE: Horses were preconditioned by exercising them on a treadmill to establish a uniform level of fitness. Each horse ran 4 simulated races, with a minimum of 14 days between races. Simulated races were run at a treadmill speed that caused horses to exercise at 120% of their maximal oxygen consumption. Horses ran until they were fatigued or for a maximum of 2 minutes. One minute after the end of exercise, horses were treated i.v. with xylazine (2.2 mg/kg of body weight), detomidine (0.04 mg/kg), a combination of xylazine (2.2 mg/kg) and acepromazine (0.04 mg/kg), or saline (0.9% NaCl) solution. Treatments were randomized so that each horse received each treatment once, in random order. Cardiopulmonary indices were measured, and samples of arterial and venous blood were collected immediately before and at specific times for 90 minutes after the end of each race. RESULTS: All sedatives produced effective sedation. The cardiopulmonary depression that was induced was qualitatively similar to that induced by administration of these sedatives to resting horses and was not severe. Sedative administration after exercise prolonged the exercise-induced increase in body temperature. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of xylazine, detomidine, or a combination of xylazine-acepromazine at twice the standard doses produced safe and effective sedation in horses that had just undergone a brief, intense bout of exercise. PMID- 10791940 TI - Effects of intravenously administered yohimbine on antinociceptive, cardiorespiratory, and postural changes induced by epidural administration of detomidine hydrochloride solution to healthy mares. AB - OBJECTIVE: To determine effects of i.v. administered yohimbine on perineal analgesia, cardiovascular and respiratory activity, and head and pelvic limb position in healthy mares following epidural administration of detomidine hydrochloride solution. ANIMALS: 8 healthy mares. PROCEDURE: Each mare received detomidine hydrochloride (0.06 mg/kg of body weight), administered in the caudal epidural space, followed 61 minutes later by yohimbine (0.05 mg/kg; test) or sterile saline (0.9% NaCl) solution (control), administered i.v., in a randomized, crossover study design with > or = 2 weeks between treatments. Analgesia was determined by lack of sensory perception to electrical stimulation of perineal dermatomes and needle-prick stimulation of coccygeal to 15th thoracic dermatomes. Arterial pH, PaCO2, PaO2, heart and respiratory rates, rectal temperature, arterial blood pressure, and cardiac output were determined, and mares were observed for sweating and urination. Mean scores obtained for test and control groups were compared. RESULTS: Intravenously administered yohimbine significantly reduced mean scores of detomidine-induced perineal analgesia, head ptosis, changes in pelvic limb position, and sweating and diuresis; antagonized detomidine-induced decreases in heart rate and cardiac output; but did not affect detomidine-induced decrease in respiratory rate. CONCLUSIONS AND CLINICAL RELEVANCE: Most effects of epidurally administered detomidine, except bradypnea, were antagonized by yohimbine, suggesting that detomidine may influence respiratory rate by mechanisms other than stimulation of alpha2-adrenoceptors, or that yohimbine induces respiratory depressant effects. Yohimbine may be an effective alpha2-adrenoceptor antagonist for all but respiratory depression following epidural administration of detomidine to mares. PMID- 10791942 TI - Ranitidine bismuth citrate and clarithromycin, alone or in combination, for eradication of Helicobacter mustelae infection in ferrets. AB - OBJECTIVE: To determine whether ranitidine bismuth citrate, clarithromycin, or a combination of ranitidine bismuth citrate and clarithromycin would be efficacious in eradication of Helicobacter mustelae infection in ferrets. ANIMALS: 60 seven month-old ferrets. PROCEDURE: To determine dosages of clarithromycin and ranitidine bismuth citrate that would suppress growth of, but not eradicate infection with, H mustelae, ferrets (n = 6/group) were treated p.o. with clarithromycin or ranitidine bismuth citrate at various dosages. Efficacy of treatment was then determined by treating ferrets with clarithromycin alone, ranitidine bismuth citrate alone, or clarithromycin and ranitidine bismuth citrate. Gastric biopsy specimens were obtained before, during, and at various times after treatment and submitted for quantitative bacterial culture and histologic evaluation. Minimum concentrations of clarithromycin that inhibited 90% of the growth of isolates obtained before and after treatment were determined. RESULTS: Dosages of clarithromycin and ranitidine bismuth citrate that suppressed growth of H mustelae were 12.5 and 24 mg/kg of body weight, p.o., every 8 hours, respectively. Infection was not eradicated in ferrets treated with ranitidine bismuth citrate alone but was eradicated in all 6 ferrets treated with clarithromycin and ranitidine bismuth citrate and in 4 of 6 treated with clarithromycin alone. A decrease in susceptibility to clarithromycin was detected for H mustelae isolates obtained after treatment. Mild or moderate antral gastritis was observed even in ferrets from which infection was eradicated. CONCLUSIONS AND CLINICAL RELEVANCE: A combination of ranitidine bismuth citrate and clarithromycin was efficacious in eradicating H mustelae infection from ferrets. PMID- 10791943 TI - Electrically elicited blink reflex in horses with trigeminal and facial nerve blocks. AB - OBJECTIVE: To reassess reference values for the components of the electrically induced blink reflex, document reference values for facial motor nerve conduction velocity, and demonstrate usefulness of the blink reflex as a diagnostic tool in peripheral facial and trigeminal nerve dysfunction in horses. ANIMALS: 10 healthy adult horses (8 males, 2 females) without neurologic abnormalities. PROCEDURE: Blink reflex tests were performed by electrical stimulation of the supraorbital nerve and facial (auriculopalpebral) nerve. Reflex and direct muscle-evoked potentials of the orbicularis oculi muscles were recorded from concentric needle electrodes inserted bilaterally in these muscles. Supraorbital and auriculopalpebral nerve blocks were performed by lidocaine hydrochloride injections. RESULTS: Supraorbital nerve stimulation elicited 2 or 3 ipsilateral and 1 contralateral reflex muscle potential in the orbicularis oculi muscles. Auriculopalpebral nerve stimulation elicited a direct and a reflex potential in the ipsilateral orbicularis oculi muscle. After left supraorbital nerve block, no responses could be elicited ipsilaterally or contralaterally upon stimulation of the blocked nerve, but bilateral responses were obtained upon stimulation of the right supraorbital nerve. After right auriculopalpebral nerve block, no responses were recorded from the right orbicularis oculi muscle upon stimulation of left or right supraorbital nerves. CONCLUSIONS AND CLINICAL RELEVANCE: Reference values for the components of the blink reflex and facial motor nerve conduction velocity will allow application of these tests to assist in the diagnosis of equine neurologic disorders involving the trigeminal and facial nerves, the brainstem, and the cranial end of the cervical segment of the spinal cord. This study reveals the usefulness of the blink reflex test in the diagnosis of peripheral trigeminal and facial nerve dysfunction in horses. PMID- 10791944 TI - Sequence of cardiovascular changes leading to pulmonary edema in swine fed culture material containing fumonisin. AB - OBJECTIVES: To determine the sequence of cardiovascular and blood gas changes induced by ingestion of fumonisin-containing culture material in swine and to examine the temporal relationship of these changes to plasma sphinganine and sphingosine concentrations. ANIMALS: 12 healthy castrated pigs (38 to 50 kg). PROCEDURE: Pigs were instrumented to permit cardiovascular monitoring and collection of blood samples. Baseline values were obtained, and pigs were randomly assigned to 1 of 2 groups. Control pigs (n = 6) were fed a standard grower diet, whereas culture material that contained 20 mg of fumonisin B1/kg of body weight was added to the feed of treated pigs (n = 6) each day. Hemodynamic data, results of arterial and mixed venous blood gas analyses, and plasma sphinganine and sphingosine concentrations were recorded every 12 hours until treated pigs were euthanatized because of impending death from pulmonary edema. RESULTS: Sphinganine and sphingosine concentrations were increased in plasma of treated pigs within 24 hours of initial fumonisin exposure and continued to increase dramatically until euthanasia. Fumonisin-treated pigs had increased respiratory rate, mean pulmonary artery pressure, and pulmonary artery wedge pressure, along with decreased heart rate and cardiac output in the 12-hour period before euthanasia. Fumonisin-treated pigs also had systemic arterial hypotension, arterial and mixed venous hypoxemia, metabolic acidosis, decreased oxygen delivery, and increased oxygen consumption immediately before euthanasia. CONCLUSIONS AND CLINICAL RELEVANCE: Fumonisin-induced pulmonary edema in swine is probably caused by acute left-sided heart failure. Onset of hemodynamic changes was associated with plasma sphinganine concentration > or = 2.2 microM/L and plasma sphingosine concentration > or = 1 microM/L. PMID- 10791945 TI - Susceptibility of equine erythrocytes to oxidant-induced rheologic alterations. AB - OBJECTIVE: To evaluate the rheologic responses of equine versus human RBC to oxidant stress induced by superoxide anions. SAMPLE POPULATION: Equine blood samples were obtained from 8 healthy, 3- to 6-year-old various breed horses of either sex; human blood samples were obtained from 8 healthy adults. PROCEDURE: Washed RBC were exposed to superoxide anions generated by the xanthine oxidase (XO)-hypoxanthine system (XO activity of 0 to 0.1 U/ml). Deformability of RBC was assessed by ektacytometry, and RBC aggregation was measured in autologous plasma or 3% solution of dextran 70 via a defined-shear photometric technique. RESULTS: Equine RBC had XO dose-dependent increases in methemoglobin concentration that were greater by 60 to 110% than in human RBC and an enhanced tendency for echinocyte formation (ie, 40% echinocyte formation at highest activity of XO). Oxidant stress reduced deformability (ie, increased rigidity) for equine and human RBC with the effect more prominent for equine RBC. Equine RBC aggregation had a biphasic response with a significant increase in plasma and dextran 70 at low XO activities and inhibition at high activities; echinocytes were incorporated into equine, but not human, RBC aggregates. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with human RBC, equine RBC are more sensitive to oxidant damage as judged by the extent of methemoglobin formation, alteration of aggregation, and reduction of cellular deformability. The high susceptibility of equine RBC to oxidant damage, and the resulting hemorheologic alterations, may have important consequences for tissue perfusion and cardiovascular adequacy in horses; they may be of particular relevance in physiologic or pathophysiologic changes associated with increased oxidant stress. PMID- 10791946 TI - Evaluation of structural and functional alterations of circulating neutrophils in calves with experimentally induced pneumonic pasteurellosis. AB - OBJECTIVES: To determine the structural and functional alterations in circulating neutrophils that may lead to sequestration in lung microvasculature and endothelial injury in calves with experimentally induced pneumonic pasteurellosis. ANIMALS: 10 healthy, 2- to 4-week-old male Holstein calves. PROCEDURES: Holstein calves were anesthetized and inoculated intrabronchially with Dulbecco phosphate buffered saline (0.9% NaCl) solution (DPBSS; 5 control calves) or 1 x 10(9) Pasteurella haemolytica organisms (5 infected calves). Blood samples were collected before and 1, 2, 4, and 6 hours after inoculation. Total and differential WBC count, dilute whole blood leukocyte deformability, neutrophil size distribution, and neutrophil surface CD11b expression were measured in blood samples. RESULTS: A progressive decrease in leukocyte deformability and increase in neutrophil size was detected 1, 2, 4, and 6 hours after inoculation of P haemolytica. Neutrophil surface CD11b expression was greater than baseline values at 6 hours after inoculation of P haemolytica. Two populations of neutrophils with an increase in size were detected in P haemolytica-infected calves. Both subpopulations had increased CD11b expression, compared with neutrophils that were typical in size. CONCLUSIONS AND CLINICAL RELEVANCE: Neutrophils circulate in an activated and nondeformable state in calves with experimentally induced pneumonic pasteurellosis. A decrease in neutrophil deformability and neutrophil aggregation may contribute to neutrophil trapping in the lung microvasculature during pneumonic pasteurellosis in calves. PMID- 10791947 TI - Evaluation of certification in the Milk and Dairy Beef Quality Assurance Program and associated factors on the risk of having violative antibiotic residues in milk from dairy farms in Michigan. AB - OBJECTIVE: To determine whether certification in a Milk and Dairy Beef Quality Assurance Program (MDBQAP) was associated with a reduced risk of having antibiotic residues in milk and to define specific management factors that may have predisposed dairy farms to having violative antibiotic residues in milk. SAMPLE POPULATION: 124 dairy farms in Michigan that had > 1 violative residue in milk during 1993 and 248 randomly selected control farms in Michigan that did not have violative residues in milk during 1993. PROCEDURE: A pretested structured questionnaire was mailed to case and control farms. A conditional multivariate logistic regression model was developed to determine risk factors associated with having a violative antibiotic residue in milk. RESULTS: Certification in the MDBQAP did not significantly reduce the risk of having a violative antibiotic residue. Annual treatment of > 10% of a herd for metritis was associated with a reduced risk of having a violative residue. Evidence suggested that a routine request for a milk processor to perform residue testing was associated with a decreased risk of having had a violative antibiotic residue, but routine on-farm residue testing was associated with an increased risk of having had a residue. CONCLUSIONS AND CLINICAL RELEVANCE: MDBQAP certification was associated, although not significantly, with a reduced risk of having violative antibiotic residues in milk. Risk factors significantly associated with violative antibiotic residues are addressed by various critical control points in the MDBQAP and may be indicators for strengths and weaknesses of MDBQAP. PMID- 10791948 TI - Corneal thickness, intraocular pressure, and optical corneal diameter in Rocky Mountain Horses with cornea globosa or clinically normal corneas. AB - OBJECTIVE: To compare corneal thickness, intraocular pressure, and optical corneal diameter in Rocky Mountain Horses with cornea globosa and those with clinically normal corneas. ANIMALS: 129 Rocky Mountain Horses. PROCEDURE: Ultrasonic pachymetry was used to measure corneal thickness. Applanation tonometry was used to measure intraocular pressure. A Jameson caliper was used to measure optical corneal diameter. RESULTS: The central and temporal peripheral portions of the cornea were significantly thicker in horses with cornea globosa than in horses with clinically normal corneas, but corneal thicknesses in the dorsal, ventral, and medial peripheral portions of the cornea were not significantly different between groups. There were no differences in corneal thickness between male and female horses or between right and left eyes. However, there was a positive correlation between age and corneal thickness. Intraocular pressure was not significantly different between horses with cornea globosa and those with clinically normal corneas, or between right and left eyes, or male and female horses. Optical corneal diameter for horses with cornea globosa was not significantly different from diameter for horses with clinically normal corneas, but optical corneal diameter was positively correlated with age. CONCLUSIONS AND CLINICAL RELEVANCE: Cornea globosa in Rocky Mountain Horses is not associated with increased intraocular pressure. Corneal thickness and optical corneal diameter increase with age in Rocky Mountain Horses. PMID- 10791949 TI - Characterization of swimming motility and identification of flagellar proteins in Salmonella pullorum isolates. AB - OBJECTIVE: To identify swimming motility in Salmonella pullorum isolates and to characterize the flagellar proteins produced by motile isolates. SAMPLE POPULATION: 30 S pullorum isolates and isolates of 7 other Salmonella sp. PROCEDURE: Salmonella pullorum isolates were inoculated into high motility medium to evaluate swimming motility. Putative flagellar proteins were purified from the organisms and analyzed by means of gel electrophoresis and western blotting procedures, using various antisera specific for flagellar proteins. Antisera shown to be reactive with putative flagellar proteins were incorporated into the growth medium to examine their effects on motility of the isolates. RESULTS: All S pullorum isolates had evidence of swimming motility. Two putative flagellar proteins were purified from 2 of the S pullorum isolates: a 60 to 62 kd protein shown to react with antiserum specific for type y flagellar protein, and a 58 to 59 kd protein shown to react with antiserum specific for type d flagellar protein and with antibody reactive to a highly conserved flagellar epitope found on various Enterobacteriaceae. Antiserum specific for type d flagellar protein inhibited swimming motility of S pullorum isolates, but antiserum specific for type y flagellar protein did not. CONCLUSIONS: Results suggest that S pullorum isolates can be induced to manifest swimming motility when grown on medium with a low agar concentration and possess a 58 to 59 kd protein of d serotype and a second protein of 60 to 62 kd that also may be a flagellar protein. PMID- 10791950 TI - Basement membrane zone type XV collagen is a disulfide-bonded chondroitin sulfate proteoglycan in human tissues and cultured cells. AB - Type XV collagen has a widespread distribution in human tissues, but a nearly restricted localization in basement membrane zones. The alpha1(XV) chain contains a highly interrupted collagenous region of 577 residues, and noncollagenous amino and carboxyl-terminal domains of 530 and 256 residues, respectively. Cysteines are present in each domain and consensus sequences for O-linked glycosaminoglycans are situated in the amino terminus and in two large, noncollagenous interruptions. We now report that type XV collagen is a chondroitin sulfate proteoglycan in human tissues and cultured cells, and that the alpha chains are covalently linked by interchain disulfide bonds only between the two cysteines in the collagenous region. Western blotting of tissue extracts revealed a diffuse smear with a mean size >/=400 kDa, which after chondroitinase digestion resolved into a 250-kDa band in umbilical cord, and 250- and 225-kDa bands in placenta, lung, colon, and skeletal muscle. The latter two bands were also directly visualized by alcian blue/silver staining of a purified placenta extract. In a human rhabdomyosarcoma cell line, almost all of the newly synthesized type XV collagen was secreted into the medium and upon chondroitinase digestion just the 250-kDa alpha chain was generated. Chondroitinase plus collagenase digestion of tissue and medium proteins followed by Western blotting using domain-specific antibodies revealed a 135-kDa amino-terminal fragment containing glycosaminoglycan chains and a 27-kDa fragment representing the intact carboxyl terminus. However, a truncated carboxyl peptide of approximately 8-kDa was also evident in tissue extracts containing the 225-kDa form. Our data suggest that the 225-kDa form arises from differential carboxyl cleavage of the 250-kDa form, and could explain the approximately 19-kDa endostatin-related fragments (John, H., Preissner, K. T., Forssmann, W.-G., and Standker, L. (1999) Biochemistry 38, 10217-10224), which may be liberated from the alpha1(XV) chain. PMID- 10791951 TI - Inhibition of the phosphoinositide 3-kinase pathway induces a senescence-like arrest mediated by p27Kip1. AB - A senescence-like growth arrest is induced in mouse primary embryo fibroblasts by inhibitors of phosphoinositide 3-kinase (PI3K). We observed that senescence-like growth arrest is correlated with an increase in p27(Kip1) but that down regulation of other cyclin-dependent kinase (CDK) inhibitors, including p15(INK4b), p16(INK4a), p19( INK4d), and p21(Cip1) as well as other negative cell cycle regulators such as p53 and p19(ARF), implies that this senescence-related growth arrest is independent of the activity of p53, p19(ARF), p16(INK4a), and p21(Cip1), which are associated with replicative senescence. The p27(Kip1) binds to the cyclin/CDK2 complexes and causes a decrease in CDK2 kinase activity. We demonstrated that ectopic expression of p27(Kip1) can induce permanent cell cycle arrest and a senescence-like phenotype in wild-type mouse embryo fibroblasts. We also obtained results suggesting that the kinase inhibitors LY294002 and Wortmannin arrest cell growth and induce a senescence-like phenotype, at least partially, through inhibition of PI3K and protein kinase B/Akt, activation of the forkhead protein AFX, and up-regulation of p27(Kip1)expression. In summary, these observations taken together suggest that p27(Kip1) is an important mediator of the permanent cell cycle arrest induced by PI3K inhibitors. Our data suggest that repression of CDK2 activity by p27(Kip1) is required for the PI3K-induced senescence, yet mouse embryo fibroblasts derived from p27(Kip1-/-) mice entered cell cycle arrest after treatment with LY294002. We show that this is due to a compensatory mechanism by which p130 functionally substitutes for the loss of p27(Kip1). This is the first description that p130 may have a role in inhibiting CDK activity during senescence. PMID- 10791952 TI - Urinary-type plasminogen activator (uPA) expression and uPA receptor localization are regulated by alpha 3beta 1 integrin in oral keratinocytes. AB - Expression of urinary-type plasminogen activator (uPA) and its receptor (uPAR) is correlated with matrix proteolysis, cell adhesion, motility, and invasion. To evaluate the functional link between adhesion and proteolysis in gingival keratinocytes (pp126), cells were treated with immobilized integrin antibodies to induce integrin clustering. Clustering of alpha(3) and beta(1) integrin subunits, but not alpha(2), alpha(5), alpha(6), or beta(4), enhanced uPA secretion. Bead immobilized laminin-5 and collagen I, two major alpha(3)beta(1) ligands, also induced uPA expression. Coordinate regulation of the serpin plasminogen activator inhibitor 1 was also apparent; however, a net increase in uPA activity was predominant. alpha(3)beta(1) integrin clustering induced extracellular signal regulated kinase 1/2 phosphorylation, and both uPA induction and extracellular signal-regulated kinase activation were blocked by the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase inhibitor PD98059. Integrin aggregation also promoted a dramatic redistribution of uPAR on the cell surface to sites of clustered alpha(3)beta(1) integrins. Co-immunoprecipitation of beta(1) integrin with uPAR provided further evidence that protein-protein interactions between uPAR and beta(1) integrin control uPAR distribution. As a functional consequence of uPA up-regulation and uPA-mediated plasminogen activation, the globular domain of the laminin-5 alpha(3) subunit, a major pp126 matrix protein, was proteolytically processed from a 190-kDa form to a 160-kDa species. Laminin-5 containing the 160-kDa alpha(3) subunit efficiently nucleates hemidesmosome formation and reduces cell motility. Together, these data suggest that multivalent aggregation of the alpha(3)beta(1) integrin regulates proteinase expression, matrix proteolysis, and subsequent cellular behavior. PMID- 10791953 TI - Smad1 and Smad4 are components of the bone morphogenetic protein-4 (BMP-4) induced transcription complex of the Xvent-2B promoter. AB - To study the mechanism of transcriptional activation of the Xenopus homeobox gene Xvent-2B, we have delineated the bone morphogenetic protein-4 (BMP-4)-responsive region between -275/-152 in the proximal promoter. Consistent with the BMP-4 inductive nature of this region, this element exhibits transcriptional activation upon ectopic expression of Smad1 and Smad4. Electrophoretic mobility shift assays with total cellular extracts demonstrated that a DNA fragment encompassing this region was competent in the formation of a BMP-4-induced protein-DNA complex containing Smad1. Two different Smad binding regions were localized, a distal binding region for Smad1 containing two GCAT motifs and proximal AGNC binding sites for Smad4, the latter being conserved in other transforming growth factor beta-responsive elements. Mutation of the Smad4 binding motif completely abolished transcriptional activation, whereas mutation or deletion of the Smad1 recognition sequence inhibited Smad1/Smad4 responsiveness. These results provide a functional characterization and identification of a vertebrate Smad1/Smad4 DNA response element induced by BMP-4 signaling and offers insight into the transcriptional regulation of a component essential for dorsoventral patterning in Xenopus embryos. PMID- 10791955 TI - MIP-T3, a novel protein linking tumor necrosis factor receptor-associated factor 3 to the microtubule network. AB - In this study, we report the identification of a novel tumor necrosis factor receptor-associated factor 3 (TRAF3)-interacting protein designated MIP-T3. MIP T3 is a 83-kDa protein with no significant homology to known mammalian proteins. MIP-T3 mRNA and TRAF3 mRNA are ubiquitously expressed, and TRAF3 is the only TRAF protein to interact with MIP-T3. The MIP-T3-TRAF3 interaction requires the coiled coil TRAF-N domain of TRAF3. To our knowledge, this is the first case of a TRAF binding protein that interacts with a single member of the TRAF family specifically through a TRAF-N coiled-coil domain. MIP-T3 binds to Taxol stabilized microtubules and to tubulin in vitro, and MIP-T3 recruits TRAF3 to microtubules when both proteins are overexpressed in HeLa cells. In a 293 cell line stably expressing CD40, TRAF3 is released from the TRAF3.MIP-T3 complex and recruited to the CD40 receptor upon CD40 ligand stimulation. MIP-T3 may provide a novel mechanism in sequestering TRAF3 to the cytoskeletal network. PMID- 10791954 TI - Concentration-dependent effects of nitric oxide on mitochondrial permeability transition and cytochrome c release. AB - The mitochondrial permeability transition pore (PTP) and associated release of cytochrome c are thought to be important in the apoptotic process. Nitric oxide (NO( small middle dot)) has been reported to inhibit apoptosis by acting on a variety of extra-mitochondrial targets. The relationship between cytochrome c release and PTP opening, and the effects of NO( small middle dot) are not clearly established. Nitric oxide, S-nitrosothiols and peroxynitrite are reported to variously inhibit or promote PTP opening. In this study the effects of NO( small middle dot) on the PTP were characterized by exposing isolated rat liver mitochondria to physiological and pathological rates of NO( small middle dot) released from NONOate NO( small middle dot) donors. Nitric oxide reversibly inhibited PTP opening with an IC(50) of 11 nm NO( small middle dot)/s, which can be readily achieved in vivo by NO( small middle dot) synthases. The mechanism involved mitochondrial membrane depolarization and inhibition of Ca(2+) accumulation. At supraphysiological release rates (>2 micrometer/s) NO( small middle dot) accelerated PTP opening. Substantial cytochrome c release occurred with only a 20% change in mitochondrial swelling, was an early event in the PTP, and was also inhibited by NO( small middle dot). Furthermore, NO( small middle dot) exposure resulted in significantly lower cytochrome c release for the same degree of PTP opening. It is proposed that this pathway represents an additional mechanism underlying the antiapoptotic effects of NO( small middle dot). PMID- 10791956 TI - Induction of secreted type IIA phospholipase A2 gene transcription by interleukin 1beta. Role of C/EBP factors. AB - Secreted type IIA phospholipase A(2), which is involved in arachidonic acid release, is abundantly produced by chondrocytes and secreted in the synovial fluids of patients affected by rheumatoid arthritis. Transfection experiments showed that interleukin-1beta stimulates the phospholipase A(2) [-1614; +20] promoter activity by 6-7-fold and that the [-210; -176] fragment is critical for this stimulation. CAAT enhancer-binding protein (C/EBP) beta and C/EBPdelta transcription factors bind to this element as shown by bandshift experiments. Interleukin-1beta increased the levels of C/EBPdelta mRNA as soon as 2 h and up to 24 h without affecting those of C/EBPbeta. Higher amounts of C/EBPdelta proteins correlate with the stimulation of C/EBPdelta mRNA. Mutations or 5' deletions in the upstream [-247; -210] region reduced by 2-fold the basal and interleukin-1beta-stimulated transcription activities. Two types of factors bind to overlapping sequences on this fragment: NF1-like proteins and the glucocorticoid receptor. The glucocorticoid receptor is responsible for a moderate stimulation of the promoter activity by dexamethasone and may interact with C/EBP factors to achieve a full transcription activity in basal conditions and in the presence of interleukin-1beta. A [-114; -85] proximal regulatory element forms three complexes in bandshift experiments, the slowest mobility one involving the Sp1 zinc finger factor. Mutation of this sequence reduced to 2-fold the stimulation of the promoter activity by interleukin-1beta or the C/EBP factors. Induction of the transcription of secreted type IIA phospholipase A(2) gene by interleukin-1beta in chondrocytes absolutely requires C/EBPbeta and C/EBPdelta factors but does not involve NF-kappaB. PMID- 10791957 TI - Biotinylation of substituted cysteines in the nicotinic acetylcholine receptor reveals distinct binding modes for alpha-bungarotoxin and erabutoxin a. AB - Although previous results indicate that alpha-subunit residues Trp(187), Val(188), Phe(189), Tyr(190), and Pro(194) of the mouse nicotinic acetylcholine receptor are solvent-accessible and are in a position to contribute to the alpha bungarotoxin (alpha-Bgtx) binding site (Spura, A., Russin, T. S., Freedman, N. D., Grant, M., McLaughlin, J. T., and Hawrot, E. (1999) Biochemistry 38, 4912 4921), little is known about the accessibility of other residues within this region. By determining second-order rate constants for the reaction of cysteine mutants at alpha184-alpha197 with the thiol-specific biotin derivative (+) biotinyl-3-maleimidopropionamidyl-3,6-dioxaoctanediamine , we now show that only very subtle differences in reactivity (approximately 10-fold) are detectable, arguing that the entire region is solvent-exposed. Importantly, biotinylation in the presence of saturating concentrations of the long neurotoxin alpha-Bgtx is significantly retarded for positions alphaW187C, alphaF189C, and reduced wild type receptors (alphaCys(192) and alphaCys(193)), further emphasizing their major contribution to the alpha-Bgtx binding site. Interestingly, although biotinylation of position alphaV188C is not affected by the presence of alpha Bgtx, erabutoxin a, which is a member of the short neurotoxin family, inhibits biotinylation at position alphaV188C, but not at alphaW187C or alphaF189C. Taken together, these results indicate that short and long neurotoxins establish interactions with distinct amino acids on the nicotinic acetylcholine receptor. PMID- 10791958 TI - Ca2+-free calmodulin and calmodulin damaged by in vitro aging are selectively degraded by 26 S proteasomes without ubiquitination. AB - The ubiquitin-proteasome pathway is believed to selectively degrade post synthetically damaged proteins in eukaryotic cells. To study this process we used calmodulin (CaM) as a substrate because of its importance in cell regulation and because it acquires isoaspartyl residues in its Ca(2+)-binding regions both in vivo and after in vitro "aging" (incubation for 2 weeks without Ca(2+)). When microinjected into Xenopus oocytes, in vitro aged CaM was degraded much faster than native CaM by a proteasome-dependent process. Similarly, in HeLa cell extracts aged CaM was degraded at a higher rate, even though it was not conjugated to ubiquitin more rapidly than the native species. Ca(2+) stimulated the ubiquitination of both species, but inhibited their degradation. Thus, for CaM, ubiquitination and proteolysis appear to be dissociated. Accordingly, purified muscle 26 S proteasomes could degrade aged CaM and native Ca(2+)-free (apo) CaM without ubiquitination. Addition of Ca(2+) dramatically reduced degradation of the native molecules but only slightly reduced the breakdown of the aged species. Thus, upon Ca(2+) binding, native CaM assumes a non-degradable conformation, which most of the age-damaged species cannot assume. Thus, flexible conformations, as may arise from age-induced damage or the absence of ligands, can promote degradation directly by the proteasome without ubiquitination. PMID- 10791960 TI - Identification of CYP4F8 in human seminal vesicles as a prominent 19-hydroxylase of prostaglandin endoperoxides. AB - A novel cytochrome P450, CYP4F8, was recently cloned from human seminal vesicles. CYP4F8 was expressed in yeast. Recombinant CYP4F8 oxygenated arachidonic acid to (18R)-hydroxyarachidonate, whereas prostaglandin (PG) D(2), PGE(1), PGE(2), PGF(2alpha), and leukotriene B(4) appeared to be poor substrates. Three stable PGH(2) analogues, 9,11-epoxymethano-PGH(2) (U-44069), 11, 9-epoxymethano-PGH(2) (U-46619), and 9,11-diazo-15-deoxy-PGH(2) (U-51605) were rapidly metabolized by omega2- and omega3-hydroxylation. U-44069 was oxygenated with a V(max) of approximately 260 pmol min(-)(1) pmol P450(-1) and a K(m) of approximately 7 micrometer. PGH(2) decomposes mainly to PGE(2) in buffer and to PGF(2alpha) by reduction with SnCl(2). CYP4F8 metabolized PGH(2) to 19-hydroxy-PGH(2), which decomposed to 19-hydroxy-PGE(2) in buffer and could be reduced to 19-hydroxy PGF(2alpha) with SnCl(2). 18-Hydroxy metabolites were also formed (approximately 17%). PGH(1) was metabolized to 19- and 18-hydroxy-PGH(1) in the same way. Microsomes of human seminal vesicles oxygenated arachidonate, U-44069, U-46619, U 51605, and PGH(2), similar to CYP4F8. (19R)-Hydroxy-PGE(1) and (19R)-hydroxy PGE(2) are the main prostaglandins of human seminal fluid. We propose that they are formed by CYP4F8-catalyzed omega2-hydroxylation of PGH(1) and PGH(2) in the seminal vesicles and isomerization to (19R)-hydroxy-PGE by PGE synthase. CYP4F8 is the first described hydroxylase with specificity and catalytic competence for prostaglandin endoperoxides. PMID- 10791959 TI - Stalk segment 4 of the yeast plasma membrane H+-ATPase. Mutational evidence for a role in the E1-E2 conformational change. AB - In the P(2)-type ATPases, there is growing evidence that four alpha-helical stalk segments connect the cytoplasmic part of the molecule, responsible for ATP binding and hydrolysis, to the membrane-embedded part that mediates cation transport. The present study has focused on stalk segment 4, which displays a significant degree of sequence conservation among P(2)-ATPases. When site directed mutants in this region of the yeast plasma membrane H(+)-ATPase were constructed and expressed in secretory vesicles, more than half of the amino acid substitutions led to a severalfold decrease in the rate of ATP hydrolysis, although they had little or no effect on the coupling between hydrolysis and transport. Strikingly, mutant ATPases bearing single substitutions of 13 consecutive residues from Ile-359 through Gly-371 were highly resistant to inorganic orthovanadate, with IC(50) values at least 10-fold above those seen in the wild-type enzyme. Most of the same mutants also displayed a significant reduction in the K(m) for MgATP and an increase in the pH optimum for ATP hydrolysis. Taken together, these changes in kinetic behavior point to a shift in equilibrium from the E(2) conformation of the ATPase toward the E(1) conformation. The residues from Ile-359 through Gly-371 would occupy three full turns of an alpha-helix, suggesting that this portion of stalk segment 4 may provide a conformationally active link between catalytic sites in the cytoplasm and cation-binding sites in the membrane. PMID- 10791961 TI - Cytochrome c methyltransferase, Ctm1p, of yeast. AB - Cytochromes c from plants and fungi, but not higher animals, contain methylated lysine residues at specific positions, including for example, the trimethylated lysine at position 72 in iso-1-cytochrome c of the yeast Saccharomyces cerevisiae. Testing of 6,144 strains of S. cerevisiae, each overproducing a different open reading frame fused to glutathione S-transferase, previously revealed that YHR109w was associated with an activity that methylated horse cytochrome c. We show here that this open reading frame, denoted Ctm1p, is specifically responsible for trimethylating lysine 72 of iso-1-cytochrome c. Unmethylated forms of cytochrome c but not other proteins or nucleic acids are methylated in vitro by Ctm1p produced in S. cerevisiae or Escherichia coli. Iso-1 cytochrome c purified from a ctm1-Delta strain is not trimethylated in vivo, whereas the K72R mutant form, or the trimethylated Lys-72 form of iso-1 cytochrome c, are not significantly methylated by Ctm1p in vitro. Like apocytochrome c, but in contrast to holocytochrome c, Ctm lp is located in the cytosol, consistent with the view that the natural substrate is apocytochrome c. The ctm1-Delta strain lacking the methyltransferase did not exhibit any growth defect on a variety of media and growth conditions, and the unmethylated iso-1 cytochrome c was produced at the normal level and exhibited the normal activity in vivo. Ctm1p and cytochrome c were coordinately regulated during anaerobic to aerobic transition, a finding consistent with the view that this methyltransferase evolved to act on cytochrome c. PMID- 10791962 TI - CaV2.2 and CaV2.3 (N- and R-type) Ca2+ channels in depolarization-evoked entry of Ca2+ into mouse sperm. AB - As sperm prepare for fertilization, surface Ca(2+) channels must open to initiate required, Ca(2+)-mediated events. However, the molecular identity and functional properties of sperm Ca(2+) channels remain uncertain. Here, we use rapid local perfusion and single-cell photometry to examine the kinetics of calcium responses of mouse sperm to depolarizing stimuli. The linear rise of intracellular [Ca(2+)] evoked by approximately 10-s applications of an alkaline high [K(+)] medium directly reports activity of voltage-gated Ca(2+) channels. Little response occurs if external Ca(2+) is removed or if external or internal pH is elevated without depolarization. Responses are inhibited 30-40% by 30-100 micrometer Ni(2+) and more completely by 100-300 micrometer Cd(2+). They resist the dihydropyridines nitrendipine and PN200-110, but 1-10 micrometer mibefradil inhibits reversibly. They also resist the venom toxins calciseptine, omega conotoxin MVIIC, and kurtoxin, but omega-conotoxin GVIA (5 micrometer) inhibits approximately 50%. GVIA also partially blocks transient, low voltage activated Ca(2+) currents of patch-clamped spermatids. Differential sensitivity of sperm responses to Ni(2+) and Cd(2+) and partial blockade by GVIA indicate that depolarization opens at least two types of voltage-gated Ca(2+) channels in epididymal sperm examined prior to capacitation. Involvement of a previously undetected Ca(V)2.2 (N-type) channel, suggested by the action of GVIA, is substantiated by immunodetection of Ca(2+) channel alpha(1B) subunits in sperm and sperm extracts. Resistance to dihydropyridines, calciseptine, MVIIC, and kurtoxin indicates that Ca(V)1, Ca(V)2.1, and Ca(V)3 (L-, P/Q-, and T-type) channels contribute little to this evoked response. Partial sensitivity to 1 micrometer mibefradil and an enhanced sensitivity of the GVIA-resistant component of response to Ni(2+) suggest participation of a Ca(V)2.3 (R-type) channel specified by previously found alpha(1E) subunits. Our examination of depolarization-evoked Ca(2+) entry indicates that mature sperm possess a larger palette of voltage-gated Ca(2+) channels than previously thought. Such diversity may permit specific responses to multiple cues encountered on the path to fertilization. PMID- 10791964 TI - Free cholesterol loading of macrophages induces apoptosis involving the fas pathway. AB - Macrophage death is an important feature of atherosclerosis, but the cellular mechanism for this process is largely unknown. There is increasing interest in cellular free cholesterol (FC) excess as an inducer of lesional macrophage death because macrophages accumulate large amounts of FC in vivo, and FC loading of macrophages in culture causes cell death. In this study, a cell culture model was used to explore the cellular mechanisms involved in the initial stages of FC induced macrophage death. After 9 h of FC loading, some of the macrophages exhibited externalization of phosphatidylserine and DNA fragmentation, indicative of an apoptotic mechanism. Incubation of the cells with Z-DEVD-fluoromethylketone blocked these events, indicating dependence upon effector caspases. Macrophages from mice with mutations in either Fas or Fas ligand (FasL) demonstrated substantial resistance to FC-induced apoptosis, and FC-induced death in wild-type macrophages was blocked by an anti-FasL antibody. FC loading had no effect on the expression of cell-surface Fas but caused a small yet reproducible increase in cell-surface FasL. To determine the physiological significance of this finding, unloaded and FC-loaded Fas-deficient macrophages, which can only present FasL, were compared for their ability to induce apoptosis in secondarily added Fas bearing macrophages. The FC-loaded macrophages were much more potent inducers of apoptosis than the unloaded macrophages, and this effect was almost completely blocked by an inhibitory anti-FasL antibody. In summary, during the early stages of FC loading of macrophages, a fraction of cells exhibited biochemical changes that are indicative of apoptosis. An important part of this event is FC-induced activation of FasL that leads to Fas-mediated apoptosis. In light of recent in vivo findings that show that apoptotic macrophages in atherosclerotic lesions express both Fas and FasL, we present a cellular model of Fas-mediated death in lesional foam cells. PMID- 10791963 TI - Cytoplasmic, nuclear, and golgi localization of RGS proteins. Evidence for N terminal and RGS domain sequences as intracellular targeting motifs. AB - RGS proteins comprise a family of proteins named for their ability to negatively regulate heterotrimeric G protein signaling. Biochemical studies suggest that members of this protein family act as GTPase-activating proteins for certain Galpha subunits, thereby accelerating the turn-off mechanism of Galpha and terminating signaling by both Galpha and Gbetagamma subunits. In the present study, we used confocal microscopy to examine the intracellular distribution of several RGS proteins in COS-7 cells expressing RGS-green fluorescent protein (GFP) fusion proteins and in cells expressing RGS proteins endogenously. RGS2 and RGS10 accumulated in the nucleus of COS-7 cells transfected with GFP constructs of these proteins. In contrast, RGS4 and RGS16 accumulated in the cytoplasm of COS-7 transfectants. As observed in COS-7 cells, RGS4 exhibited cytoplasmic localization in mouse neuroblastoma cells, and RGS10 exhibited nuclear localization in human glioma cells. Deletion or alanine substitution of an N terminal leucine repeat motif present in both RGS4 and RGS16, a domain identified as a nuclear export sequence in HIV Rev and other proteins, promoted nuclear localization of these proteins in COS-7 cells. In agreement with this observation, treatment of mouse neuroblastoma cells with leptomycin B to inhibit nuclear protein export by exportin1 resulted in accumulation of RGS4 in the nucleus of these cells. GFP fusions of RGS domains of RGS proteins localized in the nucleus, suggesting that nuclear localization of RGS proteins results from nuclear targeting via RGS domain sequences. RGSZ, which shares with RGS-GAIP a cysteine-rich string in its N-terminal region, localized to the Golgi complex in COS-7 cells. Deletion of the N-terminal domain of RGSZ that includes the cysteine motif promoted nuclear localization of RGSZ. None of the RGS proteins examined were localized at the plasma membrane. These results demonstrate that RGS proteins localize in the nucleus, the cytoplasm, or shuttle between the nucleus and cytoplasm as nucleo-cytoplasmic shuttle proteins. RGS proteins localize differentially within cells as a result of structural differences among these proteins that do not appear to be important determinants for their G protein regulating activities. These findings suggest involvement of RGS proteins in more complex cellular functions than currently envisioned. PMID- 10791965 TI - Selective stabilization of the high affinity binding conformation of glucagon receptor by the long splice variant of Galpha(s). AB - To analyze functional differences in the interactions of the glucagon receptor (GR) with the two predominant splice variants of Galpha(s), GR was covalently linked to the short and the long forms Galpha(s)-S and Galpha(s)-L to produce the fusion proteins GR-Galpha(s)-S and GR-Galpha(s)-L. GR-Galpha(s)-S bound glucagon with an affinity similar to that of GR, while GR-Galpha(s)-L showed a 10-fold higher affinity for glucagon. In the presence of GTPgammaS, GR-Galpha(s)-L reverted to the low affinity glucagon binding conformation. Both GR-Galpha(s)-L and GR-Galpha(s)-S were constitutively active, causing elevated basal levels of cAMP even in the absence of glucagon. A mutant GR that failed to activate G(s) (G23D1R) was fused to Galpha(s)-L. G23D1R-Galpha(s)-L bound glucagon with high affinity, but failed to elevate cAMP levels, suggesting that the mechanisms of GR mediated Galpha(s)-L activation and Galpha(s)-L-induced high affinity glucagon binding are independent. Both GR-Galpha(s)-S and GR-Galpha(s)-L bound the antagonist desHis(1)[Nle(9),Ala(11),Ala(16)]glucagon amide with affinities similar to GR. The antagonist displayed partial agonist activity with GR Galpha(s)-L, but not with GR-Galpha(s)-S. Therefore, the partial agonist activity of the antagonist observed in intact cells appears to be due to GRs coupled to Galpha(s)-L. We conclude that Galpha(s)-S and Galpha(s)-L interact differently with GR and that specific coupling of GR to Galpha(s)-L may account for GTP sensitive high affinity glucagon binding. PMID- 10791966 TI - Quo vadis: polarized membrane recycling in motility and phagocytosis. PMID- 10791967 TI - A kinase-anchoring protein (AKAP)95 recruits human chromosome-associated protein (hCAP)-D2/Eg7 for chromosome condensation in mitotic extract. AB - Association of the condensin multiprotein complex with chromatin is required for chromosome condensation at mitosis. What regulates condensin targeting to chromatin is largely unknown. We previously showed that the nuclear A kinase anchoring protein, AKAP95, is implicated in chromosome condensation. We demonstrate here that AKAP95 acts as a targeting protein for human chromosome associated protein (hCAP)-D2/Eg7, a component of the human condensin complex, to chromosomes. In HeLa cell mitotic extract, AKAP95 redistributes from the nuclear matrix to chromatin. When association of AKAP95 with chromatin is prevented, the chromatin does not condense. Condensation is rescued by a recombinant AKAP95 peptide containing the 306 COOH-terminal amino acids of AKAP95. Recombinant AKAP95 binds chromatin and elicits recruitment of Eg7 to chromosomes in a concentration-dependent manner. Amount of Eg7 recruited correlates with extent of chromosome condensation: resolution into distinct chromosomes is obtained only when near-endogenous levels of Eg7 are recruited. Eg7 and AKAP95 immunofluorescently colocalize to the central region of methanol-fixed metaphase chromosomes. GST pull-down data also suggest that AKAP95 recruits several condensin subunits. The results implicate AKAP95 as a receptor that assists condensin targeting to chromosomes. PMID- 10791969 TI - Keratins turn over by ubiquitination in a phosphorylation-modulated fashion. AB - Keratin polypeptides 8 and 18 (K8/18) are intermediate filament (IF) proteins that are expressed in glandular epithelia. Although the mechanism of keratin turnover is poorly understood, caspase-mediated degradation of type I keratins occurs during apoptosis and the proteasome pathway has been indirectly implicated in keratin turnover based on colocalization of keratin-ubiquitin antibody staining. Here we show that K8 and K18 are ubiquitinated based on cotransfection of His-tagged ubiquitin and human K8 and/or K18 cDNAs, followed by purification of ubiquitinated proteins and immunoblotting with keratin antibodies. Transfection of K8 or K18 alone yields higher levels of keratin ubiquitination as compared with cotransfection of K8/18, likely due to stabilization of the keratin heteropolymer. Most of the ubiquitinated species partition with the noncytosolic keratin fraction. Proteasome inhibition stabilizes K8 and K18 turnover, and is associated with accumulation of phosphorylated keratins, which indicates that although keratins are stable they still turnover. Analysis of K8 and K18 ubiquitination and degradation showed that K8 phosphorylation contributes to its stabilization. Our results provide direct evidence for K8 and K18 ubiquitination, in a phosphorylation modulated fashion, as a mechanism for regulating their turnover and suggest that other IF proteins could undergo similar regulation. These and other data offer a model that links keratin ubiquitination and hyperphosphorylation that, in turn, are associated with Mallory body deposits in a variety of liver diseases. PMID- 10791968 TI - Epsin 1 undergoes nucleocytosolic shuttling and its eps15 interactor NH(2) terminal homology (ENTH) domain, structurally similar to Armadillo and HEAT repeats, interacts with the transcription factor promyelocytic leukemia Zn(2)+ finger protein (PLZF). AB - Epsin (Eps15 interactor) is a cytosolic protein involved in clathrin-mediated endocytosis via its direct interactions with clathrin, the clathrin adaptor AP-2, and Eps15. The NH(2)-terminal portion of epsin contains a phylogenetically conserved module of unknown function, known as the ENTH domain (epsin NH(2) terminal homology domain). We have now solved the crystal structure of rat epsin 1 ENTH domain to 1.8 A resolution. This domain is structurally similar to armadillo and Heat repeats of beta-catenin and karyopherin-beta, respectively. We have also identified and characterized the interaction of epsin 1, via the ENTH domain, with the transcription factor promyelocytic leukemia Zn(2)+ finger protein (PLZF). Leptomycin B, an antifungal antibiotic, which inhibits the Crm1- dependent nuclear export pathway, induces an accumulation of epsin 1 in the nucleus. These findings suggest that epsin 1 may function in a signaling pathway connecting the endocytic machinery to the regulation of nuclear function. PMID- 10791971 TI - Nucleolar localization of human methionyl-tRNA synthetase and its role in ribosomal RNA synthesis. AB - Human aminoacyl-tRNA synthetases (ARSs) are normally located in cytoplasm and are involved in protein synthesis. In the present work, we found that human methionyl tRNA synthetase (MRS) was translocated to nucleolus in proliferative cells, but disappeared in quiescent cells. The nucleolar localization of MRS was triggered by various growth factors such as insulin, PDGF, and EGF. The presence of MRS in nucleoli depended on the integrity of RNA and the activity of RNA polymerase I in the nucleolus. The ribosomal RNA synthesis was specifically decreased by the treatment of anti-MRS antibody as determined by nuclear run-on assay and immunostaining with anti-Br antibody after incorporating Br-UTP into nascent RNA. Thus, human MRS plays a role in the biogenesis of rRNA in nucleoli, while it is catalytically involved in protein synthesis in cytoplasm. PMID- 10791970 TI - Partitioning of the 2-microm circle plasmid of Saccharomyces cerevisiae. Functional coordination with chromosome segregation and plasmid-encoded rep protein distribution. AB - The efficient partitioning of the 2-microm plasmid of Saccharomyces cerevisiae at cell division is dependent on two plasmid-encoded proteins (Rep1p and Rep2p), together with the cis-acting locus REP3 (STB). In addition, host encoded factors are likely to contribute to plasmid segregation. Direct observation of a 2-microm derived plasmid in live yeast cells indicates that the multiple plasmid copies are located in the nucleus, predominantly in clusters with characteristic shapes. Comparison to a single-tagged chromosome or to a yeast centromeric plasmid shows that the segregation kinetics of the 2-microm plasmid and the chromosome are quite similar during the yeast cell cycle. Immunofluorescence analysis reveals that the plasmid is colocalized with the Rep1 and Rep2 proteins within the yeast nucleus. Furthermore, the Rep proteins (and therefore the plasmid) tend to concentrate near the poles of the yeast mitotic spindle. Depolymerization of the spindle results in partial dispersion of the Rep proteins in the nucleus concomitant with a loosening in the association between plasmid molecules. In an ipl1-2 yeast strain, shifted to the nonpermissive temperature, the chromosomes and plasmid almost always missegregate in tandem. Our results suggest that, after DNA replication, plasmid distribution to the daughter cells occurs in the form of specific DNA-protein aggregates. They further indicate that the plasmid partitioning mechanism may exploit at least some of the components of the cellular machinery required for chromosomal segregation. PMID- 10791972 TI - Association of yeast RNA polymerase I with a nucleolar substructure active in rRNA synthesis and processing. AB - A novel ribonucleoprotein complex enriched in nucleolar proteins was purified from yeast extracts and constituents were identified by mass spectrometry. When isolated from rapidly growing cells, the assembly contained ribonucleic acid (RNA) polymerase (pol) I, and some of its transcription factors like TATA-binding protein (TBP), Rrn3p, Rrn5p, Rrn7p, and Reb1p along with rRNA processing factors, like Nop1p, Cbf5p, Nhp2p, and Rrp5p. The small nucleolar RNAs (snoRNAs) U3, U14, and MRP were also found to be associated with the complex, which supports accurate transcription, termination, and pseudouridylation of rRNA. Formation of the complex did not depend on pol I, and the complex could efficiently recruit exogenous pol I into active ribosomal DNA (rDNA) transcription units. Visualization of the complex by electron microscopy and immunogold labeling revealed a characteristic cluster-forming network of nonuniform size containing nucleolar proteins like Nop1p and Fpr3p and attached pol I. Our results support the idea that a functional nucleolar subdomain formed independently of the state of rDNA transcription may serve as a scaffold for coordinated rRNA synthesis and processing. PMID- 10791973 TI - The interaction of the chaperonin tailless complex polypeptide 1 (TCP1) ring complex (TRiC) with ribosome-bound nascent chains examined using photo-cross linking. AB - The eukaryotic chaperonin tailless complex polypeptide 1 (TCP1) ring complex (TRiC) (also called chaperonin containing TCP1 [CCT]) is a hetero-oligomeric complex that facilitates the proper folding of many cellular proteins. To better understand the manner in which TRiC interacts with newly translated polypeptides, we examined its association with nascent chains using a photo-cross-linking approach. To this end, a series of ribosome-bound nascent chains of defined lengths was prepared using truncated mRNAs. Photoactivatable probes were incorporated into these (35)S- labeled nascent chains during translation. Upon photolysis, TRiC was cross-linked to ribosome-bound polypeptides exposing at least 50-90 amino acids outside the ribosomal exit channel, indicating that the chaperonin associates with much shorter nascent chains than indicated by previous studies. Cross-links were observed for nascent chains of the cytosolic proteins actin, luciferase, and enolase, but not to ribosome-bound preprolactin. The pattern of cross-links became more complex as the nascent chain increased in length. These results suggest a chain length-dependent increase in the number of TRiC subunits involved in the interaction that is consistent with the idea that the substrate participates in subunit-specific contacts with the chaperonin. Both ribosome isolation by centrifugation through sucrose cushions and immunoprecipitation with anti-puromycin antibodies demonstrated that the photoadducts form on ribosome-bound polypeptides. Our results indicate that TRiC/CCT associates with the translating polypeptide shortly after it emerges from the ribosome and suggest a close association between the chaperonin and the translational apparatus. PMID- 10791974 TI - Caspase-2 is localized at the Golgi complex and cleaves golgin-160 during apoptosis. AB - Caspases are an extended family of cysteine proteases that play critical roles in apoptosis. Animals deficient in caspases-2 or -3, which share very similar tetrapeptide cleavage specificities, exhibit very different phenotypes, suggesting that the unique features of individual caspases may account for distinct regulation and specialized functions. Recent studies demonstrate that unique apoptotic stimuli are transduced by distinct proteolytic pathways, with multiple components of the proteolytic machinery clustering at distinct subcellular sites. We demonstrate here that, in addition to its nuclear distribution, caspase-2 is localized to the Golgi complex, where it cleaves golgin-160 at a unique site not susceptible to cleavage by other caspases with very similar tetrapeptide specificities. Early cleavage at this site precedes cleavage at distal sites by other caspases. Prevention of cleavage at the unique caspase-2 site delays disintegration of the Golgi complex after delivery of a pro apoptotic signal. We propose that the Golgi complex, like mitochondria, senses and integrates unique local conditions, and transduces pro-apoptotic signals through local caspases, which regulate local effectors. PMID- 10791975 TI - Dual role of caspase-11 in mediating activation of caspase-1 and caspase-3 under pathological conditions. AB - Caspase-11, a member of the murine caspase family, has been shown to be an upstream activator of caspase-1 in regulating cytokine maturation. We demonstrate here that in addition to its defect in cytokine maturation, caspase-11-deficient mice have a reduced number of apoptotic cells and a defect in caspase-3 activation after middle cerebral artery occlusion (MCAO), a mouse model of stroke. Recombinant procaspase-11 can autoprocess itself in vitro. Purified active recombinant caspase-11 cleaves and activates procaspase-3 very efficiently. Using a positional scanning combinatorial library method, we found that the optimal cleavage site of caspase-11 was (I/L/V/P)EHD, similar to that of upstream caspases such as caspase-8 and -9. Our results suggest that caspase-11 is a critical initiator caspase responsible for the activation of caspase-3, as well as caspase-1 under certain pathological conditions. PMID- 10791976 TI - Pro-apoptotic apoptosis protease-activating factor 1 (Apaf-1) has a cytoplasmic localization distinct from Bcl-2 or Bcl-x(L). AB - How Bcl-2 and its pro-survival relatives prevent activation of the caspases that mediate apoptosis is unknown, but they appear to act through the caspase activator apoptosis protease-activating factor 1 (Apaf-1). According to the apoptosome model, the Bcl-2-like proteins preclude Apaf-1 activity by sequestering the protein. To explore Apaf-1 function and to test this model, we generated monoclonal antibodies to Apaf-1 and used them to determine its localization within diverse cells by subcellular fractionation and confocal laser scanning microscopy. Whereas Bcl-2 and Bcl-x(L) were prominent on organelle membranes, endogenous Apaf-1 was cytosolic and did not colocalize with them, even when these pro-survival proteins were overexpressed or after apoptosis was induced. Immunogold electron microscopy confirmed that Apaf-1 was dispersed in the cytoplasm and not on mitochondria or other organelles. After the death stimuli, Bcl-2 and Bcl-x(L) precluded the release of the Apaf-1 cofactor cytochrome c from mitochondria and the formation of larger Apaf-1 complexes, which are steps that presage apoptosis. However, neither Bcl-2 nor Bcl-x(L) could prevent the in vitro activation of Apaf-1 induced by the addition of exogenous cytochrome c. Hence, rather than sequestering Apaf-1 as proposed by the apoptosome model, Bcl-2-like proteins probably regulate Apaf-1 indirectly by controlling upstream events critical for its activation. PMID- 10791977 TI - Isoforms of alpha-actinin from cardiac, smooth, and skeletal muscle form polar arrays of actin filaments. AB - We have used a positively charged lipid monolayer to form two-dimensional bundles of F-actin cross-linked by alpha-actinin to investigate the relative orientation of the actin filaments within them. This method prevents growth of the bundles perpendicular to the monolayer plane, thereby facilitating interpretation of the electron micrographs. Using alpha-actinin isoforms isolated from the three types of vertebrate muscle, i.e., cardiac, skeletal, and smooth, we have observed almost exclusively cross-linking between polar arrays of filaments, i.e., actin filaments with their plus ends oriented in the same direction. One type of bundle can be classified as an Archimedian spiral consisting of a single actin filament that spirals inward as the filament grows and the bundle is formed. These spirals have a consistent hand and grow to a limiting internal diameter of 0.4-0.7 microm, where the filaments appear to break and spiral formation ceases. These results, using isoforms usually characterized as cross-linkers of bipolar actin filament bundles, suggest that alpha-actinin is capable of cross-linking actin filaments in any orientation. Formation of specifically bipolar or polar filament arrays cross-linked by alpha-actinin may require additional factors that either determine the filament orientation or restrict the cross-linking capabilities of alpha-actinin. PMID- 10791978 TI - Drosophila beta spectrin functions independently of alpha spectrin to polarize the Na,K ATPase in epithelial cells. AB - Spectrin has been proposed to function as a sorting machine that concentrates interacting proteins such as the Na,K ATPase within specialized plasma membrane domains of polarized cells. However, little direct evidence to support this model has been obtained. Here we used a genetic approach to directly test the requirement for the beta subunit of the alphabeta spectrin molecule in morphogenesis and function of epithelial cells in Drosophila. beta Spectrin mutations were lethal during late embryonic/early larval development and they produced subtle defects in midgut morphology and stomach acid secretion. The polarized distributions of alphabeta(H) spectrin and ankyrin were not significantly altered in beta spectrin mutants, indicating that the two isoforms of Drosophila spectrin assemble independently of one another, and that ankyrin is upstream of alphabeta spectrin in the spectrin assembly pathway. In contrast, beta spectrin mutations had a striking effect on the basolateral accumulation of the Na,K ATPase. The results establish a role for beta spectrin in determining the subcellular distribution of the Na, K ATPase and, unexpectedly, this role is independent of alpha spectrin. PMID- 10791979 TI - Calcineurin activity is required for the initiation of skeletal muscle differentiation. AB - Differentiation of skeletal muscle myoblasts follows an ordered sequence of events: commitment, cell cycle withdrawal, phenotypic differentiation, and finally cell fusion to form multinucleated myotubes. The molecular signaling pathways that regulate the progression are not well understood. Here we investigate the potential role of calcium and the calcium-dependent phosphatase calcineurin in myogenesis. Commitment, phenotypic differentiation, and cell fusion are identified as distinct calcium-regulated steps, based on the extracellular calcium concentration required for the expression of morphological and biochemical markers specific to each of these stages. Furthermore, differentiation is inhibited at the commitment stage by either treatment with the calcineurin inhibitor cyclosporine A (CSA) or expression of CAIN, a physiological inhibitor of calcineurin. Retroviral-mediated gene transfer of a constitutively active form of calcineurin is able to induce myogenesis only in the presence of extracellular calcium, suggesting that multiple calcium-dependent pathways are required for differentiation. The mechanism by which calcineurin initiates differentiation includes transcriptional activation of myogenin, but does not require the participation of NFAT. We conclude that commitment of skeletal muscle cells to differentiation is calcium and calcineurin-dependent, but NFAT independent. PMID- 10791980 TI - Roles of autocrine TGF-beta receptor and Smad signaling in adipocyte differentiation. AB - TGF-beta inhibits adipocyte differentiation, yet is expressed by adipocytes. The function of TGF-beta in adipogenesis, and its mechanism of action, is unknown. To address the role of TGF-beta signaling in adipocyte differentiation, we characterized the expression of the TGF-beta receptors, and the Smads which transmit or inhibit TGF-beta signals, during adipogenesis in 3T3-F442A cells. We found that the cell-surface availability of TGF-beta receptors strongly decreased as adipogenesis proceeds. Whereas mRNA levels for Smads 2, 3, and 4 were unchanged during differentiation, mRNA levels for Smads 6 and 7, which are known to inhibit TGF-beta responses, decreased severely. Dominant negative interference with TGF-beta receptor signaling, by stably expressing a truncated type II TGF beta receptor, enhanced differentiation and decreased growth. Stable overexpression of Smad2 or Smad3 inhibited differentiation and dominant negative inhibition of Smad3 function, but not Smad2 function, enhanced adipogenesis. Increased Smad6 and Smad7 levels blocked differentiation and enhanced TGF-beta induced responses. The inhibitory effect of Smad7 on adipocyte differentiation and its cooperation with TGF-beta was associated with the C-domain of Smad7. Our results indicate that endogenous TGF-beta signaling regulates the rate of adipogenesis, and that Smad2 and Smad3 have distinct functions in this endogenous control of differentiation. Smad6 and Smad7 act as negative regulators of adipogenesis and, even though known to inhibit TGF-beta responses, enhance the effects of TGF-beta on these cells. PMID- 10791982 TI - Focal exocytosis of VAMP3-containing vesicles at sites of phagosome formation. AB - Phagocytosis involves the receptor-mediated extension of plasmalemmal protrusions, called pseudopods, which fuse at their tip to engulf a particle. Actin polymerizes under the nascent phagosome and may propel the protrusion of pseudopods. Alternatively, membrane extension could result from the localized insertion of intracellular membranes into the plasmalemma next to the particle. Here we show focal accumulation of VAMP3-containing vesicles, likely derived from recycling endosomes, in the vicinity of the nascent phagosome. Using green fluorescent protein (GFP) as both a fluorescent indicator and an exofacial epitope tag, we show that polarized fusion of VAMP3 vesicles precedes phagosome sealing. It is therefore likely that targeted delivery of endomembranes contributes to the elongation of pseudopods. In addition to mediating pseudopod formation, receptor-triggered focal secretion of endosomes may contribute to polarized membrane extension in processes such as lamellipodial elongation or chemotaxis. PMID- 10791981 TI - Analysis of notch lacking the carboxyl terminus identified in Drosophila embryos. AB - The cell surface receptor Notch is required during development of Drosophila melanogaster for differentiation of numerous tissues. Notch is often required for specification of precursor cells by lateral inhibition and subsequently for differentiation of tissues from these precursor cells. We report here that certain embryonic cells and tissues that develop after lateral inhibition, like the connectives and commissures of the central nervous system, are enriched for a form of Notch not recognized by antibodies made against the intracellular region carboxy-terminal of the CDC10/Ankyrin repeats. Western blotting and immunoprecipitation analyses show that Notch molecules lacking this region are produced during embryogenesis and form protein complexes with the ligand Delta. Experiments with cultured cells indicate that Delta promotes accumulation of a Notch intracellular fragment lacking the carboxyl terminus. Furthermore, Notch lacking the carboxyl terminus functions as a receptor for Delta. These results suggest that Notch activities during development include generation and activity of a truncated receptor we designate NDeltaCterm. PMID- 10791983 TI - Role of a new mammalian gene family in the biosynthesis of very long chain fatty acids and sphingolipids. AB - Whereas the physiological significance of microsomal fatty acid elongation is generally appreciated, its molecular nature is poorly understood. Here, we describe tissue-specific regulation of a novel mouse gene family encoding components implicated in the synthesis of very long chain fatty acids. The Ssc1 gene appears to be ubiquitously expressed, whereas Ssc2 and Cig30 show a restricted expression pattern. Their translation products are all integral membrane proteins with five putative transmembrane domains. By complementing the homologous yeast mutants, we found that Ssc1 could rescue normal sphingolipid synthesis in the sur4/elo3 mutant lacking the ability to synthesize cerotic acid (C(26:0)). Similarly, Cig30 reverted the phenotype of the fen1/elo2 mutant that has reduced levels of fatty acids in the C(20)-C(24) range. Further, we show that Ssc1 mRNA levels were markedly decreased in the brains of myelin-deficient mouse mutants known to have very low fatty acid chain elongation activity. Conversely, the dramatic induction of Cig30 expression during brown fat recruitment coincided with elevated elongation activity. Our results strongly implicate this new mammalian gene family in tissue-specific synthesis of very long chain fatty acids and sphingolipids. PMID- 10791984 TI - Direct observation of membrane tethers formed during neutrophil attachment to platelets or P-selectin under physiological flow. AB - Adhesion and subsequent aggregation between neutrophils and platelets is dependent upon the initial binding of P-selectin on activated platelets to P selectin glycoprotein ligand 1 (PSGL-1) on the microvilli of neutrophils. High speed, high resolution videomicroscopy of flowing neutrophils interacting with spread platelets demonstrated that thin membrane tethers were pulled from neutrophils in 32 +/- 4% of the interactions. After capture by spread platelets, neutrophil membrane tethers (length of 5.9 +/- 4.1 microm, n = 63) were pulled at an average rate of 6-40 microm/s as the wall shear rate was increased from 100 250 s(-1). The average tether lifetime decreased significantly (P < 0.001) from 630 to 133 ms as the shear rate was increased from 100 s(-1) (F(bond) = 86 pN) to 250 s(-1) (F(bond) = 172 pN), which is consistent with P-selectin/PSGL-1 bond dynamics under stress. Tether formation was blocked by antibodies against P selectin or PSGL-1, but not by anti-CD18 antibodies. During neutrophil rolling on P-selectin at 150 s(-1), thin membrane tethers were also pulled from the neutrophils. The characteristic jerking motion of the neutrophil coexisted with tether growth (8.9 +/- 8.8 microm long), whereas tether breakage (average lifetime of 3.79 +/- 3.32 s) caused an acute jump in the rolling velocity, proving multiple bonding in the cell surface and the tether surface contact area. Extremely long membrane tethers (>40 microm) were sometimes pulled, which detached in a flow-dependent mechanism of microparticle formation. Membrane tethers were also formed when neutrophils were perfused over platelet monolayers. These results are the first visualization of the often hypothesized tethers that shield the P-selectin/PSGL-1 bond from force loading to regulate neutrophil rolling during inflammation and thrombosis. PMID- 10791985 TI - Electromechanical coupling between skeletal and cardiac muscle. Implications for infarct repair. AB - Skeletal myoblasts form grafts of mature muscle in injured hearts, and these grafts contract when exogenously stimulated. It is not known, however, whether cardiac muscle can form electromechanical junctions with skeletal muscle and induce its synchronous contraction. Here, we report that undifferentiated rat skeletal myoblasts expressed N-cadherin and connexin43, major adhesion and gap junction proteins of the intercalated disk, yet both proteins were markedly downregulated after differentiation into myo-tubes. Similarly, differentiated skeletal muscle grafts in injured hearts had no detectable N-cadherin or connexin43; hence, electromechanical coupling did not occur after in vivo grafting. In contrast, when neonatal or adult cardiomyocytes were cocultured with skeletal muscle, approximately 10% of the skeletal myotubes contracted in synchrony with adjacent cardiomyocytes. Isoproterenol increased myotube contraction rates by 25% in coculture without affecting myotubes in monoculture, indicating the cardiomyocytes were the pacemakers. The gap junction inhibitor heptanol aborted myotube contractions but left spontaneous contractions of individual cardiomyocytes intact, suggesting myotubes were activated via gap junctions. Confocal microscopy revealed the expression of cadherin and connexin43 at junctions between myotubes and neonatal or adult cardiomyocytes in vitro. After microinjection, myotubes transferred dye to neonatal cardiomyocytes via gap junctions. Calcium imaging revealed synchronous calcium transients in cardiomyocytes and myotubes. Thus, cardiomyocytes can form electromechanical junctions with some skeletal myotubes in coculture and induce their synchronous contraction via gap junctions. Although the mechanism remains to be determined, if similar junctions could be induced in vivo, they might be sufficient to make skeletal muscle grafts beat synchronously with host myocardium. PMID- 10791987 TI - Prodrug-activating systems in suicide gene therapy. PMID- 10791988 TI - Gene delivery from replication-selective viruses: arming guided missiles in the war against cancer. PMID- 10791986 TI - Matrix survival signaling: from fibronectin via focal adhesion kinase to c-Jun NH(2)-terminal kinase. AB - Most transformed cells have lost anchorage and serum dependence for growth and survival. Previously, we established that when serum is absent, fibronectin survival signals transduced by focal adhesion kinase (FAK), suppress p53 regulated apoptosis in primary fibroblasts and endothelial cells (Ilic et al. 1998. J. Cell Biol. 143:547-560). The present goals are to identify survival sequences in FAK and signaling molecules downstream of FAK required for anchorage dependent survival of primary fibroblasts. We report that binding of the SH3 domain of p130Cas to proline-rich region 1 of FAK is required to support survival of fibroblasts on fibronectin when serum is withdrawn. The FAK-p130Cas complex activates c-Jun NH2-terminal kinase (JNK) via a Ras/Rac1/Pak1/MAPK kinase 4 (MKK4) pathway. Activated (phospho-) JNK colocalizes with FAK in focal adhesions of fibroblasts cultured on fibronectin, which supports their survival, but not in fibroblasts cultured on collagen, which does not. Cells often survive in the absence of extracellular matrix if serum factors are provided. In that case, we confirm work of others that survival signals are transduced by FAK, phosphatidylinositol 3'-kinase (PI3-kinase), and Akt/protein kinase B (PKB). However, when serum is absent, PI3-kinase and Akt/PKB are not involved in the fibronectin-FAK-JNK survival pathway documented herein. Thus, survival signals from extracellular matrix and serum are transduced by FAK via two distinct pathways. PMID- 10791989 TI - Immunomodulation of cancer: potential use of selectively replicating agents. PMID- 10791990 TI - Regulatable systems: applications in gene therapy and replicating viruses. PMID- 10791992 TI - Regulation of adrenocorticotropic hormone secretion: lessons from mice deficient in corticotropin-releasing hormone. PMID- 10791991 TI - Of metals, mice, and men: what animal models can teach us about body iron loading. PMID- 10791993 TI - A role for heterologous gap junctions between melanoma and endothelial cells in metastasis. AB - F10 and BL6 sublines of B16 mouse melanoma cells are metastatic after intravenous injection, but only BL6 cells are metastatic after subcutaneous injection. We found that connexin (Cx) 26 is upregulated in BL6 cells. To examine gap junction formation, we devised a coculture system, in which an opened vein segment was placed at the bottom of a culture dish and then dye-labeled melanoma cells were seeded onto it. Immunohistochemistry indicated that the vein segment preserved the integrity of the endothelial monolayer. In this system, BL6 cells could transfer dye into endothelial cells but F10 cells could not. Transfection with wild-type Cx26 rendered F10 cells competent for coupling with endothelial cells and as spontaneously metastatic as BL6 cells. Conversely, transfection with a dominant-negative form of Cx26 rendered BL6 cells deficient in coupling and less metastatic. In human melanoma lesions, the level of Cx26 expression was low in melanoma cells residing in the basal layer, but significantly upregulated in melanoma cells invading the dermis. The results suggested that Cx26 plays a role in intravasation and extravasation of tumor cells through heterologous gap junction formation with endothelial cells. PMID- 10791994 TI - Proinflammatory consequences of transgenic fas ligand expression in the heart. AB - Expression of Fas ligand (FasL) renders certain tissues immune privileged, but its expression in other tissues can result in severe neutrophil infiltration and tissue destruction. The consequences of enforced FasL expression in striated muscle is particularly controversial. To create a stable reproducible pattern of cardiomyocyte-specific FasL expression, transgenic (Tg) mice were generated that express murine FasL specifically in the heart, where it is not normally expressed. Tg animals are healthy and indistinguishable from nontransgenic littermates. FasL expression in the heart does result in mild leukocyte infiltration, but despite coexpression of Fas and FasL in Tg hearts, neither myocardial tissue apoptosis nor necrosis accompanies the leukocyte infiltration. Instead of tissue destruction, FasL Tg hearts develop mild interstitial fibrosis, functional changes, and cardiac hypertrophy, with corresponding molecular changes in gene expression. Induced expression of the cytokines TNF-alpha, IL-1beta, IL 6, and TGF-beta accompanies these proinflammatory changes. The histologic, functional, and molecular proinflammatory consequences of cardiac FasL expression are transgene-dose dependent. Thus, coexpression of Fas and FasL in the heart results in leukocyte infiltration and hypertrophy, but without the severe tissue destruction observed in other examples of FasL-directed proinflammation. The data suggest that the FasL expression level and other tissue-specific microenvironmental factors can modulate the proinflammatory consequences of FasL. PMID- 10791995 TI - Genes that modify the hemochromatosis phenotype in mice. AB - Hereditary hemochromatosis (HH) is a prevalent human disease caused by a mutation in HFE, which encodes an atypical HLA class I protein involved in regulation of intestinal iron absorption. To gain insight into the pathogenesis of hemochromatosis, we have bred Hfe knockout mice to strains carrying other mutations that impair normal iron metabolism. Compound mutant mice lacking both Hfe and its interacting protein, beta-2 microglobulin (B2m), deposit more tissue iron than mice lacking Hfe only, suggesting that another B2m-interacting protein may be involved in iron regulation. Hfe knockout mice carrying mutations in the iron transporter DMT1 fail to load iron, indicating that hemochromatosis involves iron flux through DMT1. Similarly, compound mutants deficient in both Hfe and hephaestin (Heph) show less iron loading than do Hfe knockout mice, indicating that iron absorption in hemochromatosis involves the function of Heph as well. Finally, compound mutants lacking Hfe and the transferrin receptor accumulate more tissue iron than do mice lacking Hfe alone, consistent with the idea that interaction between these two proteins contributes to the control of normal iron absorption. In addition to providing insight into the pathogenesis of HH, our results suggest that each of these genes might be a candidate modifier of the human hemochromatosis phenotype. PMID- 10791996 TI - Arginine, citrulline, and nitric oxide metabolism in end-stage renal disease patients. AB - The kidneys are thought to be a major site of net de novo arginine synthesis, but the quantitative status of arginine metabolism and its substrate precursor relationship to nitric oxide (NO) synthesis in end stage renal disease (ESRD) patients have not been characterized. We have investigated kinetic aspects of whole body arginine metabolism in six patients with ESRD. They received two pre- and two post-hemodialysis intravenous tracer infusion studies with L-[guanidino (15)N(2)]arginine and L-[(13)C]leucine during the first study, and L-[5 (13)C]arginine and L-[5-(13)C-ureido,5,5, (2)H(2)]citrulline during the second study. Arginine homeostasis in ESRD patients was found to be associated with a lower rate of arginine oxidation, and despite the decrease in renal function, the rate of de novo arginine synthesis appeared to be preserved. Plasma citrulline concentrations and flux were also elevated in these subjects compared with healthy adults. The rate of whole body NO synthesis was increased in the ESRD patients, but apparently not different pre- and post-hemodialysis therapy. The anatomic site(s) responsible for the maintenance of net de novo arginine synthesis and for the elevated NO synthesis and its pathophysiological importance in ESRD remain to be established. PMID- 10791997 TI - The heat-stable antigen determines pathogenicity of self-reactive T cells in experimental autoimmune encephalomyelitis. AB - Induction of myelin-specific CD4 T cells is a pivotal event in the development of experimental autoimmune encephalomyelitis (EAE). Other checkpoints in EAE pathogenesis have not been clearly defined, although multiple genetic loci are known to influence EAE development. We report here that targeted mutation of the heat-stable antigen (HSA) abrogates development of EAE despite a complete lack of effect on induction of autoimmune T cells. To test whether T-cell expression of HSA is sufficient, we created transgenic mice in which HSA is expressed exclusively in the T-cell lineage. We found that these mice remain resistant to EAE induction. Adoptive transfer studies demonstrate that both T cells and non-T cells must express HSA in order for the pathogenic T cells to execute their effector function. Moreover, HSAIg, a fusion protein consisting of the extracellular domain of the HSA and the Fc portion of immunoglobulin, drastically ameliorates the clinical sign of EAE even when administrated after self-reactive T cells had been expanded. Thus, identification of HSA as a novel checkpoint, even after activation and expansion of self-reactive T cells, provides a novel approach for immunotherapy of autoimmune neurologic diseases, such as multiple sclerosis. PMID- 10791998 TI - Epidermal growth factor receptor expression in neurofibromatosis type 1-related tumors and NF1 animal models. AB - We have found that EGF-R expression is associated with the development of the Schwann cell-derived tumors characteristic of neurofibromatosis type 1 (NF1) and in animal models of this disease. This is surprising, because Schwann cells normally lack EGF-R and respond to ligands other than EGF. Nevertheless, immunoblotting, Northern analysis, and immunohistochemistry revealed that each of 3 malignant peripheral nerve sheath tumor (MPNST) cell lines from NF1 patients expressed the EGF-R, as did 7 of 7 other primary MPNSTs, a non-NF1 MPNST cell line, and the S100(+) cells from each of 9 benign neurofibromas. Furthermore, transformed derivatives of Schwann cells from NF1(-/-) mouse embryos also expressed the EGF-R. All of the cells or cell lines expressing EGF-R responded to EGF by activation of downstream signaling pathways. Thus, EGF-R expression may play an important role in NF1 tumorigenesis and Schwann cell transformation. Consistent with this hypothesis, growth of NF1 MPNST lines and the transformed NF1(-/-) mouse embryo Schwann cells was greatly stimulated by EGF in vitro and could be blocked by agents that antagonize EGF-R function. PMID- 10791999 TI - Pathophysiological role of leptin in obesity-related hypertension. AB - To explore the pathophysiological role of leptin in obesity-related hypertension, we examined cardiovascular phenotypes of transgenic skinny mice whose elevated plasma leptin concentrations are comparable to those seen in obese subjects. We also studied genetically obese KKA(y) mice with hyperleptinemia, in which hypothalamic melanocortin system is antagonized by ectopic expression of the agouti protein. Systolic blood pressure (BP) and urinary catecholamine excretion are elevated in transgenic skinny mice relative to nontransgenic littermates. The BP elevation in transgenic skinny mice is abolished by alpha(1)-adrenergic, beta adrenergic, or ganglionic blockers at doses that do not affect BP in nontransgenic littermates. Central administration of an alpha-melanocyte stimulating hormone antagonist causes a marked increase in cumulative food intake but no significant changes in BP. The obese KKA(y) mice develop BP elevation with increased urinary catecholamine excretion relative to control KK mice. After a 2 week caloric restriction, BP elevation is reversed in nontransgenic littermates with the A(y) allele, in parallel with a reduction in plasma leptin concentrations, but is sustained in transgenic mice overexpressing leptin with the A(y) allele, which remain hyperleptinemic. This study demonstrates BP elevation in transgenic skinny mice and obese KKA(y) mice that are both hyperleptinemic, thereby suggesting the pathophysiological role of leptin in some forms of obesity-related hypertension. PMID- 10792000 TI - Adrenalectomy reduces neuropeptide Y-induced insulin release and NPY receptor expression in the rat ventromedial hypothalamus. AB - Chronic central administration of neuropeptide Y (NPY) causes hyperphagia, hyperinsulinemia, and obesity, a response that is prevented by prior adrenalectomy (ADX) in rats. The basis of NPY's effect and how the acute responses to this peptide are affected by ADX remain unknown. This study investigates the role of glucocorticoids in acute NPY-stimulated food intake, acute NPY-induced insulin release, and hypothalamic NPY-receptor mRNA expression levels. NPY-induced food intake was similar in ADX and control rats after acute intracerebroventricular injection of NPY. Injection of NPY caused a significant increase in plasma insulin in control rats, but this effect was completely absent in ADX rats in which basal plasma insulin levels were also lower than controls. In addition, ADX significantly reduced the number of neurons expressing NPY receptor Y(1) and Y(5) mRNAs in the ventromedial hypothalamus (VMH), without affecting Y(1)- or Y(5)-mRNA expression in the paraventricular hypothalamus or the arcuate nucleus. These data indicate that glucocorticoids are necessary for acute NPY-mediated insulin release and suggest that the mechanisms involve glucocorticoid regulation of Y(1) and Y(5) receptors specifically within the VMH nucleus. PMID- 10792001 TI - Regulation of expression of the multidrug resistance protein MRP1 by p53 in human prostate cancer cells. AB - The expression of several drug-resistance genes, including MRP and p53, increases with advancing stage of human prostate cancer. Altered transcription could account for the genotypic alterations associated with prostate cancer progression, and it was recently reported that the promoter of MRP1 is activated in the presence of mutant p53. To determine whether there is a relationship between p53 status and the expression of MRP1, a human, temperature-sensitive p53 mutant (tsp Val(138)) was transfected into LNCaP human prostate cancer cells. In the transfected cell line (LVCaP), the wild-type p53 produced growth arrest at the G1/S interface of the cell cycle, inhibited colony formation, and induced p21(waf1/cip1). Temperature shifting to 38 degrees C (p53 mutant) produced a time dependent increase in expression of MRP1. This change in MRP1 expression was also seen in isogenic cell lines in which p53 was inactivated by human papilloma virus (HPV)16E6 protein or by a dominant-negative mutant. Functional assays revealed a decrease in drug accumulation and drug sensitivity associated with mutant p53 and increased MRP1 expression. These results provide the first mechanistic link between expression of MRP1 and mutation of p53 in human prostate cancer and support recent clinical associations. Furthermore, these data suggest a mechanism tying accumulation of p53 mutations to the multidrug resistance phenotype seen in this disease. PMID- 10792002 TI - Corticotropin-releasing hormone links pituitary adrenocorticotropin gene expression and release during adrenal insufficiency. AB - Corticotropin-releasing hormone (CRH)-deficient (KO) mice provide a unique system to define the role of CRH in regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Despite several manifestations of chronic glucocorticoid insufficiency, basal pituitary proopiomelanocortin (POMC) mRNA, adrenocorticotrophic hormone (ACTH) peptide content within the pituitary, and plasma ACTH concentrations are not elevated in CRH KO mice. The normal POMC mRNA content in KO mice is dependent upon residual glucocorticoid secretion, as it increases in both KO and WT mice after adrenalectomy; this increase is reversed by glucocorticoid, but not aldosterone, replacement. However, the normal plasma levels of ACTH in CRH KO mice are not dependent upon residual glucocorticoid secretion, because, after adrenalectomy, these levels do not undergo the normal increase seen in KO mice despite the increase in POMC mRNA content. Administration of CRH restores ACTH secretion to its expected high level in adrenalectomized CRH KO mice. Thus, in adrenal insufficiency, loss of glucocorticoid feedback by itself can increase POMC gene expression in the pituitary; but CRH action is essential for this to result in increased secretion of ACTH. This may explain why, after withdrawal of chronic glucocorticoid treatment, reactivation of CRH secretion is a necessary prerequisite for recovery from suppression of the HPA axis. PMID- 10792004 TI - Th1 and Th2 mediate acute graft-versus-host disease, each with distinct end-organ targets. AB - STAT4 and STAT6 are transcription factors that play crucial roles in responding to IL-12 and IL-4, respectively. STAT4 gene knockout (STAT4(-/-)) mice have markedly reduced Th1 responses and enhanced Th2 responses. STAT6(-/-) mice show the inverse phenotype. We compared the ability of bone marrow transplantation (BMT) with the inclusion of spleen cells from STAT6(-/-), STAT4(-/-), and wild type (WT) mice to produce graft-versus-host disease (GVHD) in lethally irradiated MHC-mismatched recipients. Acute GVHD mortality was more rapid when induced by cells from STAT6(-/-) mice than when induced by STAT4(-/-) cells. However, cells from STAT4(-/-) and STAT6(-/-) donors both induced delayed GVHD mortality compared with WT controls, or compared with combined STAT4(-/-) and STAT6(-/-) cells, indicating a contribution of both Th1 cells and Th2 cells to acute GVHD. Recipients of STAT6(-/-) BMT showed evidence of acute GVHD with severe diarrhea and marked weight loss. Recipients of STAT4(-/-) BMT showed signs of GVHD with only initial transient weight loss and later development of severe skin GVHD. Histopathology showed that Th2 responses were required for the induction of both hepatic and severe skin GVHD. In contrast, both Th1 cells and Th2 cells were capable of causing intestinal pathology of GVHD. Our studies demonstrate an additive role for Th1 and Th2 cells in producing acute GVHD, and suggest a cytokine-directed approach to treating end-organ manifestations of GVHD. PMID- 10792003 TI - M-CSF neutralization and egr-1 deficiency prevent ovariectomy-induced bone loss. AB - Increased stromal cell production of M-CSF, an event caused by enhanced phosphorylation of the nuclear protein Egr-1, is central to the mechanism by which estrogen (E2) deficiency upregulates osteoclast (OC) formation. However, the contribution of enhanced M-CSF production to the bone loss induced by E2 deficiency remains to be determined. We found that treatment with an Ab that neutralizes M-CSF in vivo completely prevents the rise in OC number, the increase in bone resorption, and the resulting bone loss induced by ovariectomy (ovx). We also found that adult, intact Egr-1-deficient mice, a strain characterized by maximally stimulated stromal cell production of M-CSF, exhibit increased bone resorption and decreased bone mass. In these mice, treatment with anti-M-CSF Ab restored normal levels of bone resorption, thus confirming that increased M-CSF production accounts for the remodeling abnormalities of Egr-1-deficient mice. Consistent with the failure of ovx to further increase M-CSF production in Egr-1 deficient mice, ovx neither increased bone resorption further, nor caused bone loss in these animals. In summary, the data demonstrate that E2 deficiency induces M-CSF production via an Egr-1-dependent mechanism that is central to the pathogenesis of ovx-induced bone loss. Thus, Egr-1 and M-CSF are critical mediators of the bone sparing effects of E2 in vivo. PMID- 10792005 TI - Extracellular calcium elicits a chemokinetic response from monocytes in vitro and in vivo. AB - Recruitment of macrophages to sites of cell death is critical for induction of an immunologic response. Calcium concentrations in extracellular fluids vary markedly, and are particularly high at sites of injury or infection. We hypothesized that extracellular calcium participates in modulating the immune response, perhaps acting via the seven-transmembrane calcium-sensing receptor (CaR) on mature monocytes/macrophages. We observed a dose-dependent increase in monocyte chemotaxis in response to extracellular calcium or the selective allosteric CaR activator NPS R-467. In contrast, monocytes derived from mice deficient in CaR lacked the normal chemotactic response to a calcium gradient. Notably, CaR activation of monocytes bearing the receptor synergistically augmented the transmigration response of monocytes to the chemokine MCP-1 in association with increased cell-surface expression of its cognate receptor, CCR2. Conversely, stimulation of monocytes with MCP-1 or SDF-1alpha reciprocally increased CaR expression, suggesting a dual-enhancing interaction of Ca(2+) with chemokines in recruiting inflammatory cells. Subcutaneous administration in mice of Ca(2+), MCP-1, or (more potently) the combination of Ca(2+) and MCP-1, elicited an inflammatory infiltrate consisting of monocytes/macrophages. Thus extracellular calcium functions as an ionic chemokinetic agent capable of modulating the innate immune response in vivo and in vitro by direct and indirect actions on monocytic cells. Calcium deposition may be both consequence and cause of chronic inflammatory changes at sites of injury, infection, and atherosclerosis. PMID- 10792006 TI - PAC1 receptor-deficient mice display impaired insulinotropic response to glucose and reduced glucose tolerance. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a ubiquitous neuropeptide of the vasoactive intestinal peptide (VIP) family that potentiates glucose-stimulated insulin secretion. Pancreatic beta cells express two PACAP receptor subtypes, a PACAP-preferring (PAC1) and a VIP-shared (VPAC2) receptor. We have applied a gene targeting approach to create a mouse lacking the PAC1 receptor (PAC1(-/-)). These mice were viable and normoglycemic, but exhibited a slight feeding hyperinsulinemia. In vitro, in the isolated perfused pancreas, the insulin secretory response to PACAP was reduced by 50% in PAC1(-/-) mice, whereas the response to VIP was unaffected. In vivo, the insulinotropic action of PACAP was also acutely reduced, and the peptide induced impairment of glucose tolerance after an intravenous glucose injection. This demonstrates that PAC1 receptor is involved in the insulinotropic action of the peptide. Moreover, PAC1(-/-) mice exhibited reduced glucose-stimulated insulin secretion in vitro and in vivo, showing that the PAC1 receptor is required to maintain normal insulin secretory responsiveness to glucose. The defective insulinotropic action of glucose was associated with marked glucose intolerance after both intravenous and gastric glucose administration. Thus, these results are consistent with a physiological role for the PAC1 receptor in glucose homeostasis, notably during food intake. PMID- 10792008 TI - Osteopenia and decreased bone formation in osteonectin-deficient mice PMID- 10792007 TI - IL-10-producing T cells suppress immune responses in anergic tuberculosis patients. AB - The lethality of Mycobacterium tuberculosis remains the highest among infectious organisms and is linked to inadequate immune response of the host. Containment and cure of tuberculosis requires an effective cell-mediated immune response, and the absence, during active tuberculosis infection, of delayed-type hypersensitivity (DTH) responses to mycobacterial antigens, defined as anergy, is associated with poor clinical outcome. To investigate the biochemical events associated with this anergy, we screened 206 patients with pulmonary tuberculosis and identified anergic patients by their lack of dermal reactivity to tuberculin purified protein derivative (PPD). In vitro stimulation of T cells with PPD induced production of IL-10, IFN-gamma, and proliferation in PPD(+) patients, whereas cells from anergic patients produced IL-10 but not IFN-gamma and failed to proliferate in response to this treatment. Moreover, in anergic patients IL-10 producing T cells were constitutively present, and T-cell receptor-mediated (TCR mediated) stimulation resulted in defective phosphorylation of TCRzeta and defective activation of ZAP-70 and MAPK. These results show that T-cell anergy can be induced by antigen in vivo in the intact human host and provide new insights into mechanisms by which M. tuberculosis escapes immune surveillance. PMID- 10792009 TI - Loss of chromosome 14 increases the radiosensitivity of CGL1 human hybrid cells but lowers their susceptibility to radiation-induced neoplastic transformation. AB - Loss of active tumor suppressor alleles on fibroblast chromosomes 11 and 14 are involved in radiation-induced neoplastic transformation of human hybrid CGL1 cells. Loss of either chromosome 11 or 14 alone is not sufficient for neoplastic transformation. To gain insight into the potential functions of these tumor suppressor loci, we have investigated the effects of chromosome 11 or 14 loss on radiation-induced neoplastic transformation. We recently demonstrated that loss of chromosome 11 increases the susceptibility to X-ray induced cell killing, neoplastic transformation and the expression of delayed death. The data suggested that one possible function of the chromosome 11 tumor suppressor gene may be to help maintain genome stability after radiation damage. We postulated that if the chromosome 14 allele is functioning in a similar manner, then the loss of chromosome 14 may also make the hybrid cells more susceptible to radiation induced cell killing and neoplastic transformation. A hybrid cell line which has lost one copy of chromosome 14 was isolated and designated CON3(-14). CON3(-14) cells were more sensitive to X-ray-induced cell killing when compared with parental CGL1 cells. However, the susceptibility to radiation-induced neoplastic transformation was significantly reduced (by a factor of two) compared with the parental CGL1 cells. The expression of delayed death in the progeny of the irradiated CON3(-14) cells, growing in transformation flasks, was similar to CGL1 cells during the 21 day assay period. Taken together, the data indicate that loss of chromosome 14 alone increased the X-ray sensitivity of the hybrid cells but reduced their susceptibility to radiation-induced neoplastic transformation. These data suggest that the tumor suppressor alleles on chromosomes 11 and 14 may be functionally distinct in terms of their regulation of genomic instability and neoplastic transformation after radiation exposure. PMID- 10792011 TI - The bleomycin amplification assay in V79 cells predicts frameshift mutagenicity of intercalative agents. AB - We have recently reported on the use of a cell-based bleomycin amplification assay for the detection of DNA intercalating agents. In order to further validate this assay, two series of proprietary compounds were evaluated for frameshift mutagenesis in the Ames bacterial reversion system and for bleomycin amplification in the Chinese hamster V79 micronucleus system. It is shown that 10 of 11 frameshift-positive compounds were bleomycin amplifiers. These studies indicate that positive frameshift mutagenicity findings are consistent with expectations from the results of the bleomycin amplification assay, providing additional validation of the amplification assay for the detection of DNA intercalating agents. The studies also demonstrate that intercalation is necessary but not sufficient for frameshift mutagenesis since bleomycin amplifiers lacking frameshift mutagenic activity were also identified. PMID- 10792010 TI - In vitro and in vivo evaluation of the antihypertensive drug atenolol in cultured human lymphocytes: effects of long-term therapy. AB - The genotoxicity of atenolol, a beta-blocker antihypertensive drug, both in vitro and in vivo, was cytogenetically tested for its ability to induce sister chromatid exchange (SCE) and micronuclei (MN) in cultured peripheral lymphocytes. Also, fluorescence in situ hybridization (FISH) with a centromeric probe was performed to determine the origin of the induced MN. The in vivo study was carried out, on the one hand, on four patients under antihypertensive treatment with atenolol and, on the other hand, on four matched control individuals taking an oral dose of atenolol. The in vitro study was performed on the control individuals by adding the drug to the culture medium at a final concentration similar to the levels found in plasma. When a comparison was made, the frequency of SCE did not show significant differences in any case. A statistically significant increase in the frequency of MN was detected in patients but not in control individuals either in vitro or in vivo. FISH analysis revealed statistically significant differences between patients and control individuals without the drug with respect to the frequency of centromeric signals in MN. Taking all these observations together, our data suggest that chronic exposure to atenolol resulted mainly in the induction of chromosome loss, so an aneugenic activity could be predicted. Different sensitivity to the compound was observed among control individuals. Nevertheless, all of them responded to the presence of atenolol in the same way in both assays. Interindividual variability was also reported. The intervariability seen in patients suggested an adaptive response to the chemical after long-term therapy. PMID- 10792012 TI - Vanillin (3-methoxy-4-hydroxybenzaldehyde) inhibits mutation induced by hydrogen peroxide, N-methyl-N-nitrosoguanidine and mitomycin C but not (137)Cs gamma radiation at the CD59 locus in human-hamster hybrid A(L) cells. AB - We have investigated the ability of the naturally occurring plant essence vanillin (3-methoxy-4-hydroxybenzaldehyde) to inhibit mutation at the CD59 locus on human chromosome 11 by hydrogen peroxide, N-methyl-N-nitrosoguanidine, mitomycin C and (137)Cs gamma-radiation in human-hamster hybrid A(L) cells. Previous studies using vanillin have suggested that it can inhibit chromosome aberrations induced by hydrogen peroxide and mitomycin C, as well as inhibiting X ray- and UV-induced mutations at the hprt locus. Other studies with vanillin have shown that it can increase both the toxicity and mutagenicity of ethyl methane sulfonate and increase the induction of sister chromatid exchange by mitomycin C and a variety of other mutagens. The increased sensitivity of the A(L) assay, which is due in part to its ability to detect both small (single locus) and large (multilocus) genetic damage, allows us to measure the effect of vanillin at low doses of mutagen. Vanillin is shown, in these studies, to inhibit mutation induced by hydrogen peroxide, N-methyl-N-nitrosoguanidine and mitomycin C, as well as to enhance the toxicity of these agents. Vanillin had no effect on either toxicity or mutation induced by (137)Cs gamma-radiation. The vanillin-induced potentiation of H(2)O(2) toxicity is shown not to involve inhibition of catalase or glutathione peroxidase. These results show that vanillin is able to inhibit mutation at the CD59 locus and modify toxicity in a mutagen-specific manner. Possible mechanisms to explain the action of vanillin include inhibition of a DNA repair process that leads to the death of potential mutants or enhancement of DNA repair pathways that protect from mutation but create lethal DNA lesions during the repair process. PMID- 10792013 TI - Evaluation and characterization of micronuclei induced by the antitumour agent ASE [3beta-hydroxy-13alpha-amino-13, 17-seco-5alpha-androstan-17-oic-13, 17 lactam-p-bis(2-chloroethyl)amino phenylacetate] in human lymphocyte cultures. AB - 3beta - Hydroxy - 13alpha - amino - 13, 17 - seco - 5alpha - androstan - 17 -oic 13,17-lactam-p-bis(2-chloroethyl)amino phenylacetate (ASE) is a homo-aza steroidal ester of p-bis(2-chloroethyl) amino phenyl acetic acid and has been shown to display antineoplastic, mutagenic and genotoxic activity. In the present study an effort has been made to evaluate the ability of ASE to induce micronuclei (MN) in human lymphocytes treated in vitro using the cytokinesis block assay. Lympocytes were treated with different concentrations of ASE (0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 20 microg/ml) at two different cell culture times, 21 and 41 h after culture initiation. ASE treatment lasted until cell harvest, for 51 and 31 h, respectively. Two types of cultures were used, whole blood and isolated lymphocyte cultures. The content of induced MN was identified by FISH analysis, using an alpha-satellite DNA probe, in binucleate cells. Our results suggest that ASE is capable of increasing MN frequencies in human lymphocytes under both culture conditions. This increase is related to the concentration in a linear dose-dependent manner and is also dependent on the duration of treatment. FISH analysis has shown that the induced MN resulted mainly from breakage events. Additionally, a weak aneugenic effect was found at the higher concentrations in whole blood cultures as well as in isolated lymphocyte cultures. Cytotoxic effects of ASE were observed under both cell culture conditions with a linear dose-dependent relationship according to CBPI evaluation and were more pronounced in isolated lymphocyte cultures. PMID- 10792014 TI - Protection against Trp-P-2 mutagenicity by purpurin: mechanism of in vitro antimutagenesis. AB - Purpurin (1,2,4-trihydroxy-9,10-anthraquinone) is a natural pigment isolated from madder root (Rubia tinctorum) which inhibits the mutagenicity of a number of heterocyclic amines in the Ames mutagenicity test. Two effects were observed in the presence of purpurin. The rate of degradation of 3-hydroxyamino-1-methyl-5H pyrido?4,3-bindole ?Trp-P-2(NHOH) at neutral pH was increased. The major product of this purpurin-dependent degradation was identified as the parent amine 3-amino 1-methyl-5H-pyrido?4,3-bindole (Trp-P-2). Secondly, the rate of Trp-P-2 N hydroxylation, the major route of bioactivation, by PCB-treated rat hepatic microsomes was markedly decreased. Cytochrome P450-dependent O-dealkylation of methoxy-, ethoxy- and pentoxyresorufin by these microsomes was also significantly inhibited by purpurin. The nature of this inhibition was competitive. Spectrophotometric investigations suggest no direct interaction between Trp-P-2 and purpurin. Furthermore, no evidence for Trp-P-2 binding was observed with carminic acid, a structural analog of purpurin, when it was immobilized on omega aminohexyl agarose. Therefore, in vitro the proposed mechanism by which purpurin protects against heterocyclic amine-induced mutagenesis involves competitive inhibition of cytochrome P450-dependent bioactivation and accelerated degradation of the N-hydroxylamine to the parent amine. PMID- 10792015 TI - Induction of micronuclei and chromosomal aberrations by the mycotoxin patulin in mammalian cells: role of ascorbic acid as a modulator of patulin clastogenicity. AB - Patulin is a mycotoxin produced by several species of Penicillium, Aspergillus and BYSSOCHLAMYS: Patulin is a common contaminant of ripe apples used for the production of apple juice concentrates and is also present in other fruits, vegetables and food products. Patulin has been reported to have mutagenic, carcinogenic and teratogenic properties. Nevertheless, these properties are still a matter of debate. In this report, we further investigated the genotoxicity of patulin in mammalian cells by two different approaches. Firstly, we evaluated the induction of micronuclei in cytokinesis-blocked human lymphocytes. This approach is important because available data concerning the genetic toxicity of patulin in human cells is sparse. Secondly, we chose an established model for patulin genotoxicity, i.e. the chromosomal aberration assay in V79 Chinese hamster cells, to clarify whether concomitant exposure to ascorbic acid with the mycotoxin modulates or not the clastogenicity of patulin. The results unequivocally show induction of DNA-damaged cells by patulin as assessed by both cytogenetic assays. In addition, an almost complete abolition of patulin (0.8 microM) clastogenicity was observed in the presence of 80 microM ascorbic acid (P < 0.05), showing that although a genetic risk is present, ascorbic acid could somehow partially modulate this problem. PMID- 10792016 TI - Supra-additive genotoxicity of a combination of gamma-irradiation and ethyl methanesulfonate in mouse lymphoma L5178Y cells. AB - While testing for genotoxicity is usually performed on single chemicals, exposure of humans always comprises a number of genotoxic agents. The investigation of potentially synergistic effects of combinations therefore is an important issue in toxicology. Combinations of 511 keV gamma-radiation with the chemical alkylating agent ethyl methane-sulfonate were investigated in the in vitro micronucleus test in mouse lymphoma L5178Y cells. With combinations in the low dose linear effect range for the individual agents (0. 25-2 Gy and 0.8-3.2 mM, respectively), supra-additivity by 34-86% was seen. The synergism was more pronounced at the higher dose levels. Supra-additivity was confirmed in experiments using cytochalasin B and analyzing binucleate cells only, to control for putative effects on the cell cycle. Statistical significance was shown by a 2 factor analysis of variance with interaction. The results indicate that damage to DNA by gamma-radiation and alkylation could affect different rate limiting steps in the formation of micronuclei. Further investigations will have to show whether the observations are of general validity, in particular, whether other end-points of genotoxicity produce the same results and whether the degree of supra additivity is always dose dependent. The latter would have a strong impact on risk assessment for mixtures at low doses. PMID- 10792017 TI - DNA breakage in asbestos-treated normal and transformed (TSV40) rat pleural mesothelial cells. AB - Asbestos has been shown to induce cell cycle arrest, DNA repair and some abnormalities consistent with DNA damage but not DNA breakage. The purpose of the study was to investigate DNA breakage in asbestos-exposed rat pleural mesothelial cells (RPMC). RPMC were compared with their transformed counterparts, RPMC-TSV40 (i.e. p53-inactivated by infection with a retroviral recombinant encoding the SV40 large T antigen), as in the latter cells the cell cycle does not arrest and DNA repair is deficient due to ineffective p53-dependent cell cycle control. RPMC and RPMC-TSV40 were exposed to chrysotile and crocidolite asbestos and also to camptothecin for comparison. The presence of DNA breakage was determined using the single cell gel (Comet) assay with alkaline electrophoresis and quantified by measuring comet tail length (TL) and the percentage of total DNA in the tail and calculating tail moment (TM). We found that comets were generated by both types of asbestos in RPMC and in RPMC-TSV40 as well as by camptothecin in RPMC. On a per weight basis, chrysotile induced more abnormalities in comet parameters than did crocidolite. The comet TL and TM increased with fibre concentration, although less so with crocidolite than with chrysotile. When exposed to chrysotile at similar concentrations, RPMC consistently showed more abnormal comet parameters than did RPMC-TSV40. We concluded that asbestos causes DNA breakage and suggest that some of the DNA breakage measured was due to repair mechanisms in the normal RPMC. PMID- 10792018 TI - Cytogenetic monitoring of hospital workers occupationally exposed to ionizing radiation using the micronucleus centromere assay. AB - A cytogenetic study was performed in lymphocytes of hospital workers occupationally exposed to X- and gamma-rays using the micronucleus centromere assay. A comparison of the data for the exposed group and an age-matched group of non-exposed hospital workers showed a significant (P < 0.05) increase in centromere-positive micronuclei for the radiation workers, while no effect on centromere-negative micronuclei was present. The observed systematic increase in micronucleus frequency with age was mainly due to increased chromosome loss, reflected in the centromere-positivity of the micronuclei. The micronucleus frequencies were 40% higher in females than in males, which can again be attributed to higher chromosome loss. Two exposed individuals showed exceptionally high micronucleus yields, 90% of which were centromere-positive. In situ hybridization with a centromeric probe for chromosome X shows that X chromosome loss is responsible for these high micronucleus yields. In the studied population, smoking had no significant effect on the micronucleus yields. The results obtained indicate that in contrast to the predominantly clastogenic action of acute exposure to ionizing radiation, the aneugenic properties of radiation may be important after long-term chronic low dose exposure. PMID- 10792019 TI - Segregation of sex chromosomes in human lymphocytes. AB - Centromeric FISH was used to investigate the segregation of sex chromosomes in human lymphocytes. The aim of the study was to evaluate the effects of cell culture, cytokinesis block, age and sex on segregation and to compare the behaviour of the X and Y chromosomes. In uncultured T lymphocytes of five elderly women, the mean frequencies of nuclei hyperdiploid and hypodiploid for the X chromosome were not significantly affected by culturing the cells or by cytokinesis block. In cultured binucleate lymphocytes of two age groups of men, the X chromosome showed significantly higher mean frequencies of hyperdiploidy, hypodiploidy and reciprocal gain and loss than the Y chromosome. Reciprocal gain and loss of the Y chromosome was statistically significantly higher in the older than the younger men. In four women, studied in the same series, the rates of X chromosome aneuploidy did not significantly differ from those obtained in men. In conclusion, malsegregation of the X chromosome is common in lymphocytes of both men and women and more frequent than Y chromosome malsegregation. However, there is no clear sex difference for X chromosome reciprocal gain and loss. This would suggest that the high loss of the X chromosome in women, documented in metaphase studies, is due to micronucleation. PMID- 10792020 TI - Increased frequency of LOH on chromosome 9 in sporadic primary melanomas is associated with increased patient age at diagnosis. AB - We carried out statistical analysis of the frequency of loss of heterozygosity (LOH) at 10 microsatellite markers on chromosome 9. In 44 microdissected sporadic primary melanomas a comparison of LOH frequency data with other patient data, like age at diagnosis and tumour thickness, showed an interesting correlation between patient age at diagnosis and frequency of LOH on chromosome 9. The patient group with age >72 years at diagnosis (n = 22, mean age 82.3 +/- 6.0 years, mean LOH 3.4 +/- 2.3) showed significantly increased LOH frequency (OR 3.1, 95% CI 1.8-5.3; chi(2) test, P < 0.0001) compared with age group 0.05), and the absolute rate of GSH synthesis was 747 +/- 216 and 579 +/- 135 micromol x liter(-1) x day(-1), respectively (P < 0.05). Thus, a restricted dietary supply of SAA slows the rate of whole blood GSH synthesis and diminishes turnover, with maintenance of the GSH concentration in healthy subjects. PMID- 10792034 TI - The long-range supraorganization of the bacterial photosynthetic unit: A key role for PufX. AB - Bacterial photosynthesis relies on the interplay between light harvesting and electron transfer complexes, all of which are located within the intracytoplasmic membrane. These complexes capture and transfer solar energy, which is used to generate a proton gradient. In this study, we identify one of the factors that determines the organization of these complexes. We undertook a comparison of the organization of the light-harvesting complex 1 (LH1)/reaction center (RC) cores in the LH2(-) mutant of Rhodobacter sphaeroides in the presence or absence of the PufX protein. From polarized absorption spectra on oriented membranes, we conclude that PufX induces a specific orientation of the reaction center in the LH1 ring, as well as the formation of a long-range regular array of LH1-RC cores in the photosynthetic membrane. From our data, we have constructed a precise model of how the RC is positioned within the LH1 ring relative to the long (orientation) axis of the photosynthetic membrane. PMID- 10792035 TI - Signals transducers and activators of transcription (STAT)-induced STAT inhibitor 1 (SSI-1)/suppressor of cytokine signaling-1 (SOCS-1) suppresses tumor necrosis factor alpha-induced cell death in fibroblasts. AB - Signal transducers and activators of transcription (STAT)-induced STAT inhibitor 1 [SSI-1; also known as suppressor of cytokine signaling-1 (SOCS-1)] was identified as a negative feedback regulator of Janus kinase-STAT signaling. We previously generated mice lacking the SSI-1 gene (SSI-1 -/-) and showed that thymocytes and splenocytes in SSI-1 -/- mice underwent accelerated apoptosis. In this paper, we show that murine embryonic fibroblasts lacking the SSI-1 gene are more sensitive than their littermate controls to tumor necrosis factor-alpha (TNF alpha)-induced cell death. In addition, L929 cells forced to express SSI-1 (L929/SSI-1), but not SSI-3 or SOCS-5, are resistant to TNF-alpha-induced cell death. Furthermore L929/SSI-1 cells treated with TNF-alpha sustain the activation of p38 mitogen-activated protein (MAP) kinase. In contrast, SSI-1 -/- murine embryonic fibroblasts treated with TNF-alpha show hardly any activation of p38 MAP kinase. These findings suggest that SSI-1 suppresses TNF-alpha-induced cell death, which is mediated by p38 MAP kinase signaling. PMID- 10792036 TI - Aberrant B cell receptor signaling from B29 (Igbeta, CD79b) gene mutations of chronic lymphocytic leukemia B cells. AB - Chronic lymphocytic leukemia (CLL) B cells characteristically exhibit low or undetectable surface B cell receptor (BCR) and diminished responses to BCR mediated signaling. These features suggest that CLL cells may have sustained mutations affecting one or more of the BCR proteins required for receptor surface assembly and signal transduction. Loss of expression and mutations in the critical BCR protein B29 (Igbeta, CD79b), are prevalent in CLL and could produce the hallmark features of these leukemic B cells. Because patient CLL cells are intractable to manipulation, we developed a model system to analyze B29 mutations. Jurkat T cells stably expressing micro, kappa, and mb1 efficiently assembled a functional BCR when infected with recombinant vaccinia virus bearing wild-type B29. In contrast, a B29 CLL mutant protein truncated in the transmembrane domain did not associate with mu or mb1 at the cell surface. Another B29 CLL mutant lacking the C-terminal immunoreceptor tyrosine activation motif tyrosine and distal residues brought the receptor to the surface as well as wild-type B29 but showed significant impairment in anti-IgM-stimulated signaling events including mitogen-activated protein kinase activation. These findings demonstrate that B29 mutations previously identified in CLL patients can affect BCR-dependent signaling and may contribute to the unresponsive B cell phenotype in CLL. Finally, the features of the B29 mutations in CLL predict that they may be generated by somatic hypermutation. PMID- 10792038 TI - Separation of the glucose-stimulated cytoplasmic and mitochondrial NAD(P)H responses in pancreatic islet beta cells. AB - Two-photon excitation microscopy was used to image and quantify NAD(P)H autofluorescence from intact pancreatic islets under glucose stimulation. At maximal glucose stimulation, the rise in whole-cell NAD(P)H levels was estimated to be approximately 30 microM. However, because glucose-stimulated insulin secretion involves both glycolytic and Kreb's cycle metabolism, islets were cultured on extracellular matrix that promotes cell spreading and allows spatial resolution of the NAD(P)H signals from the cytoplasm and mitochondria. The metabolic responses in these two compartments are shown to be differentially stimulated by various nutrient applications. The glucose-stimulated increase of NAD(P)H fluorescence within the cytoplasmic domain is estimated to be approximately 7 microM. Likewise, the NAD(P)H increase of the mitochondrial domain is approximately 60 microM and is delayed with respect to the change in cytoplasmic NAD(P)H by approximately 20 sec. The large mitochondrial change in glucose-stimulated NAD(P)H thus dominates the total signal but may depend on the smaller but more rapid cytoplasmic increase. PMID- 10792037 TI - Conversion of a c type cytochrome to a b type that spontaneously forms in vitro from apo protein and heme: implications for c type cytochrome biogenesis and folding. AB - Cytochrome c(552) from Hydrogenobacter thermophilus, a thermophilic bacterium, has been converted into a b type cytochrome, after mutagenesis of both heme binding cysteines to alanine and expression in the cytoplasm of Escherichia coli. The b type variant is less stable, with the guanidine hydrochloride unfolding midpoint occurring at a concentration 2 M lower than for the wild-type protein. The reduction potential is 75 mV lower than that of the recombinant wild-type protein. The heme can be removed from the b type variant, thus generating an apo protein that has, according to circular dichroism spectroscopy, an alpha-helical content different from that of the holo b type protein. The latter is readily reformed in vitro by addition of heme to the apo protein. This reforming suggests that previously observed assembly of cytochrome c(552), which has the typical class I cytochrome c fold, in the E. coli cytoplasm is a consequence of spontaneous thioether bond formation after binding of heme to a prefolded polypeptide. These observations have implications for the general problem of c type cytochrome biogenesis. PMID- 10792039 TI - Bepridil opens the regulatory N-terminal lobe of cardiac troponin C. AB - Cardiac troponin C (cTnC) is the calcium-dependent switch for contraction in heart muscle and a potential target for drugs in the therapy of congestive heart failure. This calmodulin-like protein consists of two lobes connected by a central linker; each lobe contains two EF-hand domains. The regulatory N-terminal lobe of cTnC, unlike that of skeletal troponin C (sTnC), contains only one functional EF-hand and does not open fully upon the binding of Ca(2+). We have determined the crystal structure of cTnC, with three bound Ca(2+) ions, complexed with the calcium-sensitizer bepridil, to 2.15-A resolution. In contrast to apo- and 3Ca(2+)-cTnC, the drug-bound complex displays a fully open N-terminal lobe similar to the N-terminal lobes of 4Ca(2+)-sTnC and cTnC bound to a C-terminal fragment of cardiac troponin I (residues 147-163). The closing of the lobe is sterically hindered by one of the three bound bepridils. Our results provide a structural basis for the Ca(2+)-sensitizing effect of bepridil and reveal the details of a distinctive two-stage mechanism for Ca(2+) regulation by troponin C in cardiac muscle. PMID- 10792040 TI - Inverse radiation dose-rate effects on somatic and germ-line mutations and DNA damage rates. AB - The mutagenic effect of low linear energy transfer ionizing radiation is reduced for a given dose as the dose rate (DR) is reduced to a low level, a phenomenon known as the direct DR effect. Our reanalysis of published data shows that for both somatic and germ-line mutations there is an opposite, inverse DR effect, with reduction from low to very low DR, the overall dependence of induced mutations being parabolically related to DR, with a minimum in the range of 0.1 to 1.0 cGy/min (rule 1). This general pattern can be attributed to an optimal induction of error-free DNA repair in a DR region of minimal mutability (MMDR region). The diminished activation of repair at very low DRs may reflect a low ratio of induced ("signal") to spontaneous background DNA damage ("noise"). Because two common DNA lesions, 8-oxoguanine and thymine glycol, were already known to activate repair in irradiated mammalian cells, we estimated how their rates of production are altered upon radiation exposure in the MMDR region. For these and other abundant lesions (abasic sites and single-strand breaks), the DNA damage rate increment in the MMDR region is in the range of 10% to 100% (rule 2). These estimates suggest a genetically programmed optimatization of response to radiation in the MMDR region. PMID- 10792041 TI - Computation, prediction, and experimental tests of fitness for bacteriophage T7 mutants with permuted genomes. AB - We created a simulation based on experimental data from bacteriophage T7 that computes the developmental cycle of the wild-type phage and also of mutants that have an altered genome order. We used the simulation to compute the fitness of more than 10(5) mutants. We tested these computations by constructing and experimentally characterizing T7 mutants in which we repositioned gene 1, coding for T7 RNA polymerase. Computed protein synthesis rates for ectopic gene 1 strains were in moderate agreement with observed rates. Computed phage-doubling rates were close to observations for two of four strains, but significantly overestimated those of the other two. Computations indicate that the genome organization of wild-type T7 is nearly optimal for growth: only 2.8% of random genome permutations were computed to grow faster, the highest 31% faster, than wild type. Specific discrepancies between computations and observations suggest that a better understanding of the translation efficiency of individual mRNAs and the functions of qualitatively "nonessential" genes will be needed to improve the T7 simulation. In silico representations of biological systems can serve to assess and advance our understanding of the underlying biology. Iteration between computation, prediction, and observation should increase the rate at which biological hypotheses are formulated and tested. PMID- 10792042 TI - Misactivated amino acids translocate at similar rates across surface of a tRNA synthetase. AB - Certain aminoacyl-tRNA synthetases have a second active site that destroys (by hydrolysis) errors of amino acid activation. For example, isoleucyl-tRNA synthetase misactivates valine (to produce valyl adenylate or Val-tRNA(Ile)) at its active site. The misactivated amino acid is then translocated to an editing site located >25 A away. The role of the misactivated amino acid in determining the rate of translocation is not known. Valyl-tRNA synthetase, a close homolog of isoleucyl-tRNA synthetase, misactivates threonine, alpha-aminobutyrate, and cysteine. In this paper, we use a recently developed fluorescence-energy-transfer assay to study translocation of misactivated threonine, alpha-aminobutyrate, and cysteine. Although their rates of misactivation are clearly distinct, their rates of translocation are similar. Thus, the rate of translocation is independent of the nature of the misactivated amino acid. This result suggests that the misactivated amino acid per se has little or no role in directing translocation. PMID- 10792043 TI - Rapid transition in the structure of a coral reef community: the effects of coral bleaching and physical disturbance. AB - Coral reef communities are in a state of change throughout their geographical range. Factors contributing to this change include bleaching (the loss of algal symbionts), storm damage, disease, and increasing abundance of macroalgae. An additional factor for Caribbean reefs is the aftereffects of the epizootic that reduced the abundance of the herbivorous sea urchin, Diadema antillarum. Although coral reef communities have undergone phase shifts, there are few studies that document the details of such transitions. We report the results of a 40-month study that documents changes in a Caribbean reef community affected by bleaching, hurricane damage, and an increasing abundance of macroalgae. The study site was in a relatively pristine area of the reef surrounding the island of San Salvador in the Bahamas. Ten transects were sampled every 3-9 months from November 1994 to February 1998. During this period, the corals experienced a massive bleaching event resulting in a significant decline in coral abundance. Algae, especially macroalgae, increased in abundance until they effectively dominated the substrate. The direct impact of Hurricane Lili in October 1996 did not alter the developing community structure and may have facilitated increasing algal abundance. The results of this study document the rapid transition of this reef community from one in which corals and algae were codominant to a community dominated by macroalgae. The relatively brief time period required for this transition illustrates the dynamic nature of reef communities. PMID- 10792044 TI - Single-molecule protein folding: diffusion fluorescence resonance energy transfer studies of the denaturation of chymotrypsin inhibitor 2. AB - We report single-molecule folding studies of a small, single-domain protein, chymotrypsin inhibitor 2 (CI2). CI2 is an excellent model system for protein folding studies and has been extensively studied, both experimentally (at the ensemble level) and theoretically. Conformationally assisted ligation methodology was used to synthesize the proteins and site-specifically label them with donor and acceptor dyes. Folded and denatured subpopulations were observed by fluorescence resonance energy transfer (FRET) measurements on freely diffusing single protein molecules. Properties of these subpopulations were directly monitored as a function of guanidinium chloride concentration. It is shown that new information about different aspects of the protein folding reaction can be extracted from such subpopulation properties. Shifts in the mean transfer efficiencies are discussed, FRET efficiency distributions are translated into potentials, and denaturation curves are directly plotted from the areas of the FRET peaks. Changes in stability caused by mutation also are measured by comparing pseudo wild-type CI2 with a destabilized mutant (K17G). Current limitations and future possibilities and prospects for single-pair FRET protein folding investigations are discussed. PMID- 10792045 TI - Rapid regulated dense-core vesicle exocytosis requires the CAPS protein. AB - Although many proteins essential for regulated neurotransmitter and peptide hormone secretion have been identified, little is understood about their precise roles at specific stages of the multistep pathway of exocytosis. To study the function of CAPS (Ca(2+)-dependent activator protein for secretion), a protein required for Ca(2+)-dependent exocytosis of dense-core vesicles, secretory responses in single rat melanotrophs were monitored by patch-clamp membrane capacitance measurements. Flash photolysis of caged Ca(2+) elicited biphasic capacitance increases consisting of rapid and slow components with distinct Ca(2+) dependencies. A threshold of approximately 10 microM Ca(2+) was required to trigger the slow component, while the rapid capacitance increase was recorded already at a intracellular Ca(2+) activity < 10 microM. Both kinetic membrane capacitance components were abolished by botulinum neurotoxin B or E treatment, suggesting involvement of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)-dependent vesicle fusion. The rapid but not the slow component was inhibited by CAPS antibody. These results were further clarified by immunocytochemical studies that revealed that CAPS was present on only a subset of dense-core vesicles. Overall, the results indicate that dense-core vesicle exocytosis in melanotrophs occurs by two parallel pathways. The faster pathway exhibits high sensitivity to Ca(2+) and requires the presence of CAPS, which appears to act at a late stage in the secretory pathway. PMID- 10792046 TI - Cyclin A activates the DNA polymerase delta -dependent elongation machinery in vitro: A parvovirus DNA replication model. AB - Replication of the single-stranded linear DNA genome of parvovirus minute virus of mice (MVM) starts with complementary strand synthesis from the 3'-terminal snap-back telomere, which serves as a primer for the formation of double-stranded replicative form (RF) DNA. This DNA elongation reaction, designated conversion, is exclusively dependent on cellular factors. In cell extracts, we found that complementary strand synthesis was inhibited by the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) and rescued by the addition of proliferating cell nuclear antigen, arguing for the involvement of DNA polymerase (Pol) delta in the conversion reaction. In vivo time course analyses using synchronized MVM-infected A9 cells allowed initial detection of MVM RF DNA at the G(1)/S phase transition, coinciding with the onset of cyclin A expression and cyclin A-associated kinase activity. Under in vitro conditions, formation of RF DNA was efficiently supported by A9 S cell extracts, but only marginally by G(1) cell extracts. Addition of recombinant cyclin A stimulated DNA conversion in G(1) cell extracts, and correlated with a concomitant increase in cyclin A-associated kinase activity. Conversely, a specific antibody neutralizing cyclin A-dependent kinase activity, abolished the capacity of S cell extracts for DNA conversion. We found no evidence for the involvement of cyclin E in the regulation of the conversion reaction. We conclude that cyclin A is necessary for activation of complementary strand synthesis, which we propose as a model reaction to study the cell cycle regulation of the Pol delta-dependent elongation machinery. PMID- 10792047 TI - Phosphorylation of protein kinase N by phosphoinositide-dependent protein kinase 1 mediates insulin signals to the actin cytoskeleton. AB - Growth factors such as insulin regulate phosphatidylinositol 3-kinase-dependent actin cytoskeleton rearrangement in many types of cells. However, the mechanism by which the insulin signal is transmitted to the actin cytoskeleton remains largely unknown. Yeast two-hybrid screening revealed that the phosphatidylinositol 3-kinase downstream effector phosphoinositide-dependent protein kinase-1 (PDK1) interacted with protein kinase N (PKN), a Rho-binding Ser/Thr protein kinase potentially implicated in a variety of cellular events, including phosphorylation of cytoskeletal components. PDK1 and PKN interacted in vitro and in intact cells, and this interaction was mediated by the kinase domain of PDK1 and the carboxyl terminus of PKN. In addition to a direct interaction, PDK1 also phosphorylated Thr(774) in the activation loop and activated PKN. Insulin treatment or ectopic expression of the wild-type PDK1 or PKN, but not protein kinase Czeta, induced actin cytoskeleton reorganization and membrane ruffling in 3T3-L1 fibroblasts and Rat1 cells that stably express the insulin receptor (Rat1-IR). However, the insulin-stimulated actin cytoskeleton reorganization in Rat1-IR cells was prevented by expression of kinase-defective PDK1 or PDK1-phosphorylation site-mutated PKN. Thus, phosphorylation by PDK1 appears to be necessary for PKN to transduce signals from the insulin receptor to the actin cytoskeleton. PMID- 10792048 TI - Mapping the interface between calmodulin and MARCKS-related protein by fluorescence spectroscopy. AB - MARCKS-related protein (MRP) is a myristoylated protein kinase C substrate that binds calmodulin (CaM) with nanomolar affinity. To obtain structural information on this protein, we have engineered 10 tryptophan residues between positions 89 and 104 in the effector domain, a 24-residue-long amphipathic segment that mediates binding of MRP to CaM. We show that the effector domain is in a polar environment in free MRP, suggesting exposure to water, in agreement with a rod shaped structure of the protein. The effector domain participates in the binding of MRP to CaM, as judged by the dramatic changes observed in the fluorescent properties of the mutants on complex formation. Intermolecular quenching of the fluorescence emission of the tryptophan residues in MRP by selenomethionine residues engineered in CaM reveals that the N-terminal side of the effector domain contacts the C-terminal domain of CaM, whereas the C-terminal side of the effector domain contacts the N-terminal domain of CaM. Finally, a comparison of the fluorescent properties of the myristoylated and unmyristoylated forms of a construct in which a tryptophan residue was introduced at position 4 close to the myristoylated N terminus of MRP suggests that the lipid moiety is also involved in the interaction of MRP with CaM. PMID- 10792049 TI - A universal scaling law between gray matter and white matter of cerebral cortex. AB - Neocortex, a new and rapidly evolving brain structure in mammals, has a similar layered architecture in species over a wide range of brain sizes. Larger brains require longer fibers to communicate between distant cortical areas; the volume of the white matter that contains long axons increases disproportionally faster than the volume of the gray matter that contains cell bodies, dendrites, and axons for local information processing, according to a power law. The theoretical analysis presented here shows how this remarkable anatomical regularity might arise naturally as a consequence of the local uniformity of the cortex and the requirement for compact arrangement of long axonal fibers. The predicted power law with an exponent of 4/3 minus a small correction for the thickness of the cortex accurately accounts for empirical data spanning several orders of magnitude in brain sizes for various mammalian species, including human and nonhuman primates. PMID- 10792050 TI - The tomato ethylene receptors NR and LeETR4 are negative regulators of ethylene response and exhibit functional compensation within a multigene family. AB - The plant hormone ethylene is involved in many developmental processes, including fruit ripening, abscission, senescence, and leaf epinasty. Tomato contains a family of ethylene receptors, designated LeETR1, LeETR2, NR, LeETR4, and LeETR5, with homology to the Arabidopsis ETR1 ethylene receptor. Transgenic plants with reduced LeETR4 gene expression display multiple symptoms of extreme ethylene sensitivity, including severe epinasty, enhanced flower senescence, and accelerated fruit ripening. Therefore, LeETR4 is a negative regulator of ethylene responses. Reduced expression of this single gene affects multiple developmental processes in tomato, whereas in Arabidopsis multiple ethylene receptors must be inactivated to increase ethylene response. Transgenic lines with reduced NR mRNA levels exhibit normal ethylene sensitivity but elevated levels of LeETR4 mRNA, indicating a functional compensation of LeETR4 for reduced NR expression. Overexpression of NR in lines with lowered LeETR4 gene expression eliminates the ethylene-sensitive phenotype, indicating that despite marked differences in structure these ethylene receptors are functionally redundant. PMID- 10792051 TI - Estimating the probability for a protein to have a new fold: A statistical computational model. AB - Structural genomics aims to solve a large number of protein structures that represent the protein space. Currently an exhaustive solution for all structures seems prohibitively expensive, so the challenge is to define a relatively small set of proteins with new, currently unknown folds. This paper presents a method that assigns each protein with a probability of having an unsolved fold. The method makes extensive use of protomap, a sequence-based classification, and scop, a structure-based classification. According to protomap, the protein space encodes the relationship among proteins as a graph whose vertices correspond to 13,354 clusters of proteins. A representative fold for a cluster with at least one solved protein is determined after superposition of all scop (release 1.37) folds onto protomap clusters. Distances within the protomap graph are computed from each representative fold to the neighboring folds. The distribution of these distances is used to create a statistical model for distances among those folds that are already known and those that have yet to be discovered. The distribution of distances for solved/unsolved proteins is significantly different. This difference makes it possible to use Bayes' rule to derive a statistical estimate that any protein has a yet undetermined fold. Proteins that score the highest probability to represent a new fold constitute the target list for structural determination. Our predicted probabilities for unsolved proteins correlate very well with the proportion of new folds among recently solved structures (new scop 1.39 records) that are disjoint from our original training set. PMID- 10792052 TI - The crystal structure of the Rev binding element of HIV-1 reveals novel base pairing and conformational variability. AB - The crystal and molecular structure of an RNA duplex corresponding to the high affinity Rev protein binding element (RBE) has been determined at 2.1-A resolution. Four unique duplexes are present in the crystal, comprising two structural variants. In each duplex, the RNA double helix consists of an annealed 12-mer and 14-mer that form an asymmetric internal loop consisting of G-G and G-A noncanonical base pairs and a flipped-out uridine. The 12-mer strand has an A form conformation, whereas the 14-mer strand is distorted to accommodate the bulges and noncanonical base pairing. In contrast to the NMR model of the unbound RBE, an asymmetric G-G pair with N2-N7 and N1-O6 hydrogen bonding, is formed in each helix. The G-A base pairing agrees with the NMR structure in one structural variant, but forms a novel water-mediated pair in the other. A backbone flip and reorientation of the G-G base pair is required to assume the RBE conformation present in the NMR model of the complex between the RBE and the Rev peptide. PMID- 10792053 TI - Preselection of retrovirally transduced bone marrow avoids subsequent stem cell gene silencing and age-dependent extinction of expression of human beta-globin in engrafted mice. AB - Transcriptional silencing of genes transferred into hematopoietic stem cells poses one of the most significant challenges to the success of gene therapy. If the transferred gene is not completely silenced, a progressive decline in gene expression as the mice age often is encountered. These phenomena were observed to various degrees in mouse transplant experiments using retroviral vectors containing a human beta-globin gene, even when cis-linked to locus control region derivatives. Here, we have investigated whether ex vivo preselection of retrovirally transduced stem cells on the basis of expression of the green fluorescent protein driven by the CpG island phosphoglycerate kinase promoter can ensure subsequent long-term expression of a cis-linked beta-globin gene in the erythroid lineage of transplanted mice. We observed that 100% of mice (n = 7) engrafted with preselected cells concurrently expressed human beta-globin and the green fluorescent protein in 20-95% of their RBC for up to 9.5 mo posttransplantation, the longest time point assessed. This expression pattern was successfully transferred to secondary transplant recipients. In the presence of beta-locus control region hypersensitive site 2 alone, human beta-globin mRNA expression levels ranged from 0.15% to 20% with human beta-globin chains detected by HPLC. Neither the proportion of positive blood cells nor the average expression levels declined with time in transplanted recipients. Although suboptimal expression levels and heterocellular position effects persisted, in vivo stem cell gene silencing and age-dependent extinction of expression were avoided. These findings support the further investigation of this type of vector for the gene therapy of human hemoglobinopathies. PMID- 10792054 TI - Establishment and persistence of photoperiodic memory in hamsters. AB - Long summer days unequivocally stimulate, and short winter days inhibit reproduction in Siberian hamsters. By contrast, intermediate-duration day lengths (12.5-14 h long) either accelerate reproductive development or initiate regression of the reproductive apparatus. Which of these outcomes transpires depends on an animal's photoperiodic history, suggesting that hamsters must encode a representation of prior photoperiods. The duration of nocturnal melatonin secretion is the endocrine representation of day length, but nothing is known about how long it takes to establish photoperiodic histories or how long they endure. Hamsters exposed for 2 or more weeks to long summer day lengths acquired a long-day photoperiodic history that determined subsequent reproductive responses to intermediate-duration day lengths and melatonin signals. The memory for long-day lengths persisted in pinealectomized hamsters for 6.5 weeks, faded significantly after 13 weeks, and was functionally absent after 20 weeks. These findings indicate that hamsters are influenced only by relatively recent day lengths and melatonin signals and ignore earlier ones that might cause them to misinterpret the salience of current day lengths. PMID- 10792056 TI - Automated parallel DNA sequencing on multiple channel microchips. AB - We report automated DNA sequencing in 16-channel microchips. A microchip prefilled with sieving matrix is aligned on a heating plate affixed to a movable platform. Samples are loaded into sample reservoirs by using an eight-tip pipetting device, and the chip is docked with an array of electrodes in the focal plane of a four-color scanning detection system. Under computer control, high voltage is applied to the appropriate reservoirs in a programmed sequence that injects and separates the DNA samples. An integrated four-color confocal fluorescent detector automatically scans all 16 channels. The system routinely yields more than 450 bases in 15 min in all 16 channels. In the best case using an automated base-calling program, 543 bases have been called at an accuracy of >99%. Separations, including automated chip loading and sample injection, normally are completed in less than 18 min. The advantages of DNA sequencing on capillary electrophoresis chips include uniform signal intensity and tolerance of high DNA template concentration. To understand the fundamentals of these unique features we developed a theoretical treatment of cross-channel chip injection that we call the differential concentration effect. We present experimental evidence consistent with the predictions of the theory. PMID- 10792055 TI - Deficits in memory and motor performance in synaptotagmin IV mutant mice. AB - Synaptotagmin (Syt) IV is a synaptic vesicle protein. Syt IV expression is induced in the rat hippocampus after systemic kainic acid treatment. To examine the functional role of this protein in vivo, we derived Syt IV null [Syt IV(-/-)] mutant mice. Studies with the rotorod revealed that the Syt IV mutants have impaired motor coordination, a result consistent with constitutive Syt IV expression in the cerebellum. Because Syt IV is thought to modulate synaptic function, we also have examined Syt IV mutant mice in learning and memory tests. Our studies show that the Syt IV mutation disrupts contextual fear conditioning, a learning task sensitive to hippocampal and amygdala lesions. In contrast, cued fear conditioning is normal in the Syt IV mutants, suggesting that this mutation did not disrupt amygdala function. Conditioned taste aversion, which also depends on the amygdala, is normal in the Syt IV mutants. Consistent with the idea that the Syt IV mutation preferentially affects hippocampal function, Syt IV mutant mice also display impaired social transmission of food preference. These studies demonstrate that Syt IV is critical for brain function and suggest that the Syt IV mutation affects hippocampal-dependent learning and memory, as well as motor coordination. PMID- 10792057 TI - Noise in neurons is message dependent. AB - Neuronal responses are conspicuously variable. We focus on one particular aspect of that variability: the precision of action potential timing. We show that for common models of noisy spike generation, elementary considerations imply that such variability is a function of the input, and can be made arbitrarily large or small by a suitable choice of inputs. Our considerations are expected to extend to virtually any mechanism of spike generation, and we illustrate them with data from the visual pathway. Thus, a simplification usually made in the application of information theory to neural processing is violated: noise is not independent of the message. However, we also show the existence of error-correcting topologies, which can achieve better timing reliability than their components. PMID- 10792059 TI - Myogenic stem cell function is impaired in mice lacking the forkhead/winged helix protein MNF. AB - Myocyte nuclear factor (MNF) is a winged helix transcription factor that is expressed selectively in myogenic stem cells (satellite cells) of adult animals. Using a gene knockout strategy to generate a functional null allele at the Mnf locus, we observed that mice lacking MNF are viable, but severely runted. Skeletal muscles of Mnf-/- animals are atrophic, and satellite cell function is impaired. Muscle regeneration after injury is delayed and incomplete, and the normal timing of expression of cell cycle regulators and myogenic determination genes is dysregulated. Mnf mutant mice were intercrossed with mdx mice that lack dystrophin and exhibit only a subtle myopathic phenotype. In contrast, mdx mice that also lack MNF die in the first few weeks of life with a severe myopathy. Haploinsufficiency at the Mnf locus (Mnf+/-) also exacerbates the mdx phenotype to more closely resemble Duchenne's muscular dystrophy in humans. We conclude that MNF acts to regulate genes that coordinate the proliferation and differentiation of myogenic stem cells after muscle injury. Animals deficient in MNF may prove useful for evaluation of potential therapeutic interventions to promote muscle regeneration for patients having Duchenne's muscular dystrophy. PMID- 10792062 TI - Postnatal management of antenatal hydronephrosis using an observational approach. PMID- 10792058 TI - A neuronal beta subunit (KCNMB4) makes the large conductance, voltage- and Ca2+ activated K+ channel resistant to charybdotoxin and iberiotoxin. AB - Large conductance voltage and Ca(2+)-activated K(+) (MaxiK) channels couple intracellular Ca(2+) with cellular excitability. They are composed of a pore forming alpha subunit and modulatory beta subunits. The pore blockers charybdotoxin (CTx) and iberiotoxin (IbTx), at nanomolar concentrations, have been invaluable in unraveling MaxiK channel physiological role in vertebrates. However in mammalian brain, CTx-insensitive MaxiK channels have been described [Reinhart, P. H., Chung, S. & Levitan, I. B. (1989) Neuron 2, 1031-1041], but their molecular basis is unknown. Here we report a human MaxiK channel beta subunit (beta4), highly expressed in brain, which renders the MaxiK channel alpha subunit resistant to nanomolar concentrations of CTx and IbTx. The resistance of MaxiK channel to toxin block, a phenotype conferred by the beta4 extracellular loop, results from a dramatic ( approximately 1,000 fold) slowdown of the toxin association. However once bound, the toxin block is apparently irreversible. Thus, unusually high toxin concentrations and long exposure times are necessary to determine the role of "CTx/IbTx-insensitive" MaxiK channels formed by alpha + beta4 subunits. PMID- 10792063 TI - Antenatal hydronephrosis and ureteropelvic junction obstruction: the case for early intervention. PMID- 10792060 TI - A functional R domain from cystic fibrosis transmembrane conductance regulator is predominantly unstructured in solution. AB - Phosphorylation of the regulatory (R) domain initiates cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel activity. To discover how the function of this domain is determined by its structure, we produced an R domain protein (R8) that spanned residues 708-831 of CFTR. Phosphorylated, but not unphosphorylated, R8 stimulated activity of CFTR channels lacking this domain, indicating that R8 is functional. Unexpectedly, this functional R8 was predominantly random coil, as revealed by CD and limited proteolysis. The CD spectra of both phosphorylated and nonphosphorylated R8 were similar in aqueous buffer. The folding agent trimethylamine N-oxide induced only a small increase in the helical content of nonphosphorylated R8 and even less change in the helical content of phosphorylated R8. These data, indicating that the R domain is predominantly random coil, may explain the seemingly complex way in which phosphorylation regulates CFTR channel activity. PMID- 10792064 TI - The Monti procedure: applications and complications. PMID- 10792067 TI - Editorial comment PMID- 10792065 TI - Prostate-specific membrane antigen: present and future applications. PMID- 10792066 TI - Laparoscopic radical prostatectomy: preliminary results. AB - OBJECTIVES: To evaluate our preliminary experience with laparoscopic radical prostatectomy. The indications for laparoscopy are currently being extended to complex oncologic procedures. METHODS: Forty-three men underwent laparoscopic radical prostatectomy. We used five trocars. The surgical technique replicates the steps of traditional retropubic prostatectomy, except that the rectoprostatic cleavage plane is developed transperitoneally at the beginning of the procedure. In the first 10 patients, we performed the vesicourethral reconstruction with interrupted sutures; in the remaining 33 patients, we performed it with two hemicircumferential running sutures. The specimen was removed through the umbilical port site. RESULTS: Once the developmental phase with the first 10 patients was concluded, the median operating time was 4.3 hours without pelvic lymphadenectomy, and the median postoperative bladder catheterization was 4 days. Two (4.7%) of 43 patients underwent transfusion. Twelve patients (27.9%) had positive surgical margins; all patients had a postoperative prostate-specific antigen level of less than 0.1 ng/mL at 1 month. Rectal injury occurred in 1 patient, requiring colostomy, and 4 patients had urethrovesical anastomotic leakages requiring surgical repair. One month postoperatively, 36 patients (84%) were fully continent (no leakage). Six patients had had erections, and four stated they had had sexual intercourse. CONCLUSIONS: Laparoscopic radical prostatectomy has evolved to a fully standardized and reproducible procedure. The short-term oncologic and functional efficacy rates are equivalent to those for open surgery. The operating time is reasonable once the learning curve is over, and postoperative morbidity is diminished. Because of the improved visual accuracy, permitting more precise dissection, this technique has the potential to become an important advancement in urologic surgery. PMID- 10792068 TI - Thymosin beta-15 predicts for distant failure in patients with clinically localized prostate cancer-results from a pilot study. AB - OBJECTIVES: To report the results of a pilot study on the prognostic value of a newly identified actin-binding protein, thymosin beta-15 (Tbeta15), in predicting prostate-specific antigen (PSA) and bone failure in patients with Gleason 6/10 clinically localized prostate cancer. METHODS: Thirty-two patients (median age 70 years) with clinically localized, moderately differentiated (Gleason 6/10) prostate cancer treated by external beam radiotherapy alone (68.4 Gy) with available paraffin blocks at the Massachusetts General Hospital were evaluated for this pilot study. All patients had clinical Stage M0 disease at initial presentation, which was documented by bone scan (T1c-4,NX). Their corresponding biopsy specimens were stained immunohistochemically for Tbeta15, which was then correlated with the clinical outcome in a blinded manner. The median follow-up was 6 years (range 1 to 19) for all of the patients. RESULTS: The outcomes of the 32 patients can be grouped into three categories: patients with no evidence of disease (n = 11), patients with PSA failure without documented bone failure (n = 11), and patients with PSA failure and documented bone failure (n = 10). Tbeta15 staining intensity strongly correlated with clinical outcome. Of those patients whose specimens stained 3+ (strongest staining), 62% developed bone failure compared with 13% of those patients whose specimens stained 1+ (weakest staining) (P = 0.01). The 5-year freedom from PSA failure was only 25% for those patients with 3+ staining compared with 83% for those with 1+ staining (P = 0.02). CONCLUSIONS: The results of this pilot study have demonstrated that Tbeta15 staining intensity may be a potentially important marker to identify high-risk patients with moderately differentiated, clinically localized prostate cancer. PMID- 10792069 TI - Evaluation of an implant that delivers leuprolide for 1 year for the palliative treatment of prostate cancer. AB - OBJECTIVES: To evaluate the Viadur implant, which delivers leuprolide acetate for the palliative treatment of advanced prostate cancer. METHODS: Inserted subcutaneously, the 4 x 45-mm implant uses osmotic pressure to deliver leuprolide continuously at a controlled rate for 1 year. This 19-center open-label study enrolled patients with prostate cancer who had had no prior therapy or showed biochemical evidence of treatment failure after prostatectomy or radiotherapy. Each patient received one implant. After 1 year, that implant was removed, another was inserted, and patients were followed up for 2 additional months. The primary efficacy measure was suppression of testosterone to less than the castrate threshold (50 ng/dL). RESULTS: Eighty patients were enrolled. The implant effectively suppressed testosterone in 79 patients (99%) within 2 to 4 weeks and maintained that suppression through the study period. In 1 patient, the testosterone was suppressed to less than 100 ng/dL within 4 weeks but was not less than 50 ng/dL until week 24. Prostate-specific antigen levels normalized (4 ng/mL or less) or a clinically significant decrease occurred in all patients. Leuprolide was rapidly absorbed, resulting in mean serum concentrations of 16.8 ng/mL 4 hours after implant insertion and 2.4 ng/mL at 24 hours; steady mean serum leuprolide concentrations were then maintained throughout the year, at approximately 0.9 ng/mL. Investigators were satisfied with the insertion and removal procedures. All patients reported satisfaction after 1 year of treatment. The safety profile of the implant was consistent with androgen ablation therapy. Most adverse events were mild, and the most common event was hot flashes. CONCLUSIONS: The leuprolide implant effectively suppressed testosterone concentrations to less than the castrate threshold and maintained that suppression throughout the study period. PMID- 10792070 TI - Percutaneous sacral third nerve root neurostimulation improves symptoms and normalizes urinary HB-EGF levels and antiproliferative activity in patients with interstitial cystitis. AB - OBJECTIVES: A highly effective treatment for interstitial cystitis (IC) remains elusive. We determined whether sacral third nerve root (S3) percutaneous neurostimulation (PNS) might be effective in relieving symptoms of IC, as well as in normalizing urinary factors that are specifically altered in IC. METHODS: Six consecutive patients with symptoms and cystoscopic findings compatible with IC underwent 5 days of continuous S3 neurostimulation by way of leads placed percutaneously into the S3 foramen. Patients filled out voiding frequency diaries and pain and urgency questionnaires before PNS and at the end of PNS when the leads were removed. Urine specimens were collected at these two time points and measured for heparin-binding epidermal growth factor-like growth factor (HB-EGF) by enzyme-linked immunosorbent assay and for antiproliferative factor (APF) activity by (3)H-thymidine uptake by normal human bladder urothelial cells. RESULTS: S3 PNS significantly improved all measured parameters toward normal values. Voiding frequency decreased twofold from 23.1 +/- 4.6 to 10.6 +/- 4.0 voids daily during PNS (P = 0.0001). Pelvic pain on a scale of 1 to 10 decreased from 7.0 +/- 1.6 to 2.3 +/- 3.2 (P = 0.05). Urinary urgency on a scale of 1 to 10 decreased from 6.0 +/- 2.2 to 1.8 +/- 1.7 (P = 0. 02). Urinary HB-EGF concentration increased sevenfold from 1.5 +/- 2. 1 to 11.0 +/- 1.7 ng/mL (P <0.0001), and urinary APF activity decreased from -76.1% +/- 31% to -4.5% +/- 8.8% (P = 0.005). CONCLUSIONS: S3 PNS significantly decreased symptoms and normalized urinary HB-EGF and APF activity in patients with IC. These results suggest that permanent S3 PNS may be beneficial in treating IC. PMID- 10792071 TI - Initial assessment of a new preparatory tool for board certification in urology. AB - OBJECTIVES: To establish a Canadian national examination simulating the qualifying Urology examinations of the Royal College of Physicians and Surgeons of Canada (RCPSC) and to survey candidate perceptions regarding the mock examination's utility as a preparatory tool for the RCPSC examinations and the adequacy of examination preparation provided by their residency training program. METHODS: From January 1997 to February 1999, the Queen's Urology Examination Skills Training (QUEST) program was established, consisting of a short answer question component and an objective structured clinical examination. All participants and residency program directors received detailed feedback regarding candidate performance. RESULTS: All 11 Canadian training programs were represented, for a total of 64 participants. The 56 final-year candidates participating in QUEST and the RCPSC examinations in the same calendar year represented 66% of Canadian residents attempting the RCPSC examinations during that period. QUEST participants were surveyed immediately after the RCPSC examinations (response rate 84%). Of the respondents, 96% believed that QUEST was representative of the RCPSC examinations, and 100% said they would recommend it to future residents. Most respondents (92%) believed that the QUEST program improved their performance on the RCPSC examinations. Regarding the examination preparation received, 32 (68%) of 47 believed their program provided inadequate preparation for the RCPSC examinations. CONCLUSIONS: Support is strong among Canadian urology residents for a preparatory examination such as the QUEST program. A significant majority of residents believed that they received inadequate preparation for the RCPSC examinations from their residency training program. PMID- 10792073 TI - Editorial comment PMID- 10792072 TI - Endourologic management of benign ureteral strictures with and without compromised vascular supply. AB - OBJECTIVES: To retrospectively assess the efficacy of balloon dilation, endopyelotomy/ureterotomy, and stenting alone in the management of benign ureteral strictures with intact or compromised vascular supply. METHODS: One hundred fourteen patients with benign ureteral strictures were assessed after at least a 2-year follow-up (range 2 to 16 years, mean 6.3). Balloon dilation was performed in 81, endopyelotomy/ureterotomy with temporary stenting in 27, and ureteral stenting alone in 6 patients. Ureteral strictures were divided into strictures with intact or with compromised vascular supply. RESULTS: Balloon dilation was successful in short ureteral strictures with intact vascular supply in 33 of 37 (89.2%), but only in 3 of 8 (37.5%) long ureteral strictures and in 1 of 2 (50%) recurrent ureteropelvic junction strictures. Balloon dilation was less successful when the vascular supply was compromised in 2 (40%) of 5 short strictures, 1 (16.7%) of 6 long strictures, and 2 (33.3%) of 6 recurrent ureteropelvic junction strictures. Endopyelotomy/ureterotomy was successful in 17 (89.5%) of 19 strictures with compromised vascular supply. CONCLUSIONS: Balloon dilation is recommended for management of short strictures with intact vascular supply. Endoureterotomy with stenting is recommended for all long ureteral strictures, for ureteropelvic junction stenoses, and for short ureteral strictures with compromised vascular supply and benign underlying etiology. PMID- 10792074 TI - Using technetium-99M dimercaptosuccinic acid renal cortex scintigraphy to differentiate acute pyelonephritis from other causes of fever in patients with spinal cord injury. AB - OBJECTIVES: To differentiate acute pyelonephritis (APN) from fever due to other sources in patients with spinal cord injury by using technetium-99m dimercaptosuccinic acid (DMSA) renal cortex scintigraphy (DMSA scan). METHODS: A total of 24 patients with spinal cord injury were admitted with fever. DMSA scans were performed on all patients. The final determination of the cause of the fever was based on the medical history, physical examination, laboratory evaluation, and imaging studies. RESULTS: DMSA scan accurately diagnosed APN in 12 patients with inflammation on the DMSA scan. In 5 patients with scarred lesions on the DMSA scans and in 7 patients with negative DMSA scan, the fever was attributed to other causes. The sensitivity and specificity of the DMSA scan for detecting APN were both 100%. CONCLUSIONS: The DMSA scan is a valuable adjunct in the evaluation of fever in patients with SCI. PMID- 10792076 TI - Predicting extravesical extension of bladder carcinoma: a novel method based on micrometer measurement of the depth of invasion in transurethral resection specimens. AB - OBJECTIVES: Patients with bladder cancer and extravesical extension (Stage T3 or greater) have worse survival than those with organ-confined cancer. We sought to determine whether the depth of invasion in transurethral resection of the bladder (TURB) specimens will predict extravesical extension in patients treated by radical cystectomy. METHODS: We studied 90 patients diagnosed with invasive bladder carcinoma between 1979 and 1984. The 1997 TNM (tumor, lymph node, metastasis) system was used for pathologic staging. The mean patient age was 65 years (range 44 to 78). The male/female ratio was 5:1. All patients had invasive bladder cancer at TURB. Muscle invasion was identified in 35 patients (39%) and lamina propria invasion was present in 55 patients (61%) in the TURB specimens. The depth of invasion in the TURB specimens was measured by an ocular micrometer. All patients were treated by radical cystectomy. The median interval from TURB to cystectomy was 44 days (range 2 to 159). Extravesical extension (Stage T3 or greater) at cystectomy was present in 39 patients (43%). RESULTS: The depth of invasion was associated with final pathologic stage (Spearman correlation r = 0. 58, P <0.001). The overall accuracy of the depth of invasion for the prediction of extravesical extension, measured by the area under the receiver operating characteristic curve, was 0.81 (standard error 0. 045). The mean depth of invasion among patients with extravesical extension at cystectomy was 4.0 mm compared with 2.2 mm for those without extravesical extension. On the basis of a 4.0-mm cutoff point, the sensitivity, specificity, positive predictive value, and negative predictive value for extravesical extension were 54%, 90%, 81%, and 72%, respectively. CONCLUSIONS: Patients with a bladder cancer depth of invasion greater than 4 mm in the TURB specimens, as measured by micrometer, are likely to have extravesical extension, and more aggressive treatment should be considered. PMID- 10792075 TI - Risk factors for the development of bladder cancer after upper tract urothelial cancer. AB - OBJECTIVES: To determine the clinical and pathologic risk factors for initial intravesical recurrence in patients with primary renal pelvic and/or ureteral cancer and to examine the progression in the bladder in patients having high risk factors for intravesical recurrence. METHODS: This study included 69 patients with renal pelvic and/or ureteral cancer. We excluded patients with distant metastases, those with a short period of follow-up, and those having a previous history or concomitance of bladder cancer. The exclusion criteria were chosen to avoid contamination by patients with a poor prognosis who might die of the primary cancer before bladder cancer development. Multivariate analysis by Cox's proportional hazards model was used to determine what clinical and pathologic variables significantly affected the initial intravesical recurrence of cancer. We also studied the stage progression of cancer that recurred in the bladder. RESULTS: Initial intravesical recurrence of the cancer was found in 22 patients during a median follow-up period of 53 months (range 12 to 225). The intravesical disease-free rate after upper tract urothelial cancer was 65% (rate of disease recurrence in bladder 35%) at 5 years by the Kaplan-Meier method. The extent (multifocality) of the upper urinary cancer (P = 0.0038) and pathologic stage (P = 0.0409) independently influenced intravesical recurrence. Age, sex, adjuvant chemotherapy, configuration of the primary tumor, primary cancer size, and pathologic grade did not affect recurrence. The rate of stage progression also was not influenced by the extent of the disease in the upper urinary tract. CONCLUSIONS: The extent and pathologic stage of cancer in the upper urinary tract were significant and independent factors for initial intravesical recurrence of cancer. However, no difference was found in clinical outcome in terms of stage progression between patients having high risk factors for intravesical recurrence and those without them. PMID- 10792077 TI - Long-term results of intravesical bacillus Calmette-Guerin therapy for stage T1 superficial bladder cancer. AB - OBJECTIVES: To examine in a prospective study the incidence of recurrence and progression in patients with Stage T1 bladder carcinoma after complete transurethral resection of the bladder tumor and adjuvant immunotherapy with bacillus Calmette-Guerin (BCG). METHODS: Between July 1987 and April 1999, 126 patients presenting to our clinic with a superficial urothelial carcinoma of the bladder (Stage pT1, grade 1-3) received adjuvant intravesical immunotherapy with BCG after complete transurethral resection of the bladder tumor. In the case of recurrence of superficial tumor (pTa, pT1, or carcinoma in situ), patients received a second cycle of BCG. For muscle-invasive tumor progression (pT2, pT3, or pT4), radical cystectomy was recommended. Six of the patients (5%) presented with Stage pT1,G1 tumor, 74 (59%) with Stage pT1,G2 tumor, and 46 patients (36%) with Stage pT1,G3 tumor. Median follow-up was 53 months (range 3 to 144). RESULTS: One hundred eight patients (86%) remained tumor-free with a retained bladder during the follow-up after one or two 6-week cycles of BCG. Twenty-four patients (19%) had a recurrence of superficial tumor, 13 (10%) had muscle invasive progression after the first BCG cycle, and an additional 4 (3%) had progression after the second BCG cycle. Six patients (5%) underwent radical cystectomy, and 9 patients (7%) died as a result of tumor progression. The tumor free survival rate of all patients was 89% (112 of 126). CONCLUSIONS: Adjuvant immunotherapy with BCG after complete transurethral resection of the bladder tumor represents a highly effective primary treatment for Stage T1 carcinoma of the bladder. Even in Stage pT1,G3 tumor, immediate radical cystectomy does not appear necessary. PMID- 10792078 TI - C-ERBB-2 gene amplification as a prognostic marker in human bladder cancer. AB - OBJECTIVES: To evaluate c-erbB-2 gene amplification and its prognostic significance in transitional cell carcinoma of the bladder. METHODS: Alterations in the gene copy number of c-erbB-2 were detected in 57 bladder tumor samples using a method based on the polymerase chain reaction. RESULTS: Eighteen tumors (32%) showed gene amplification of c-erbB-2, which correlated with tumor grade and stage. A strong association of c-erbB-2 amplification with patient survival was also found. The amplification resulted in a significantly poorer prognosis among the patients with high-grade and/or invasive tumors. Multivariate analysis revealed that c-erbB-2 amplification and tumor grade were independent prognostic factors. CONCLUSIONS: Our data indicate a possible role of the c-erbB-2 gene in the development of aggressive behavior in bladder tumors. Moreover, the use of c erbB-2 gene amplification, together with tumor grade and stage, could provide an accurate basis for determining the prognosis of bladder cancer. PMID- 10792079 TI - Pretreatment with finasteride decreases perioperative bleeding associated with transurethral resection of the prostate. AB - OBJECTIVES: The efficacy of finasteride in the treatment of gross hematuria associated with benign prostatic hyperplasia is well established. We evaluated a regimen of pretreatment with finasteride in decreasing perioperative bleeding associated with transurethral resection of the prostate (TURP). METHODS: A prospective analysis compared 25 patients pretreated with finasteride for 2 to 4 months before TURP with 50 patients without pretreatment. Patients in each group were further separated by the amount of prostate tissue resected. Patients were then followed up for perioperative bleeding, defined as a perioperative blood transfusion requirement or a return visit to the emergency room with gross hematuria or clot retention. RESULTS: None of the patients with less than 30 g of prostate tissue resected experienced perioperative bleeding. In patients with 30 g or more resected, several episodes of bleeding occurred. In the patients pretreated with finasteride, 1 (8.3%) of 12 experienced perioperative bleeding; in the control group, 7 (36.8%) of 19 had an episode of bleeding. CONCLUSIONS: In patients with large prostate glands undergoing TURP, pretreatment with finasteride appears useful in reducing perioperative bleeding. PMID- 10792080 TI - Editorial comment PMID- 10792081 TI - Reply by the authors PMID- 10792082 TI - Use of a prostate model to assist in training for digital rectal examination. AB - OBJECTIVES: To assess the accuracy of prostate size estimation on digital rectal examination (DRE) before and after training with a three-dimensional prostate model relative to prostate volume by transrectal ultrasound (TRUS). METHODS: A total of 100 subjects underwent DRE by one of four family physicians (FP1, n = 34; FP2, n = 26; FP3, n = 22; and FP4, n = 18). One half were examined before any training on DRE prostate size examination and one half after the physicians were trained. Training involved teaching with a three-dimensional prostate model having posterior surface areas corresponding to the average dimensions of six different prostate volumes. The FPs were instructed to estimate the prostate size on the DRE to the nearest 5 g. A single urologist unaware of the DRE results performed TRUS on all patients to measure the prostate volume. RESULTS: Before training, the DRE size estimates ranged from 10 to 100 g (mean +/- SD 32.8 +/- 21.6), with a TRUS volume of 11 to 122 g (mean +/- SD 38.9 +/- 23.1). The correlation between the DRE and TRUS estimates was 0.25, suggesting low agreement (intraclass correlation coefficient [ICC] 0.35, 95% confidence interval 0.31, 0. 38). After training, 50 different patients had DRE size estimates of 10 to 100 g (mean +/- SD 39.4 +/- 19.7) and TRUS volume of 10 to 119 g (mean +/- SD 41.5 +/- 24.1). The correlation between the techniques was higher in patients examined after training (r = 0. 765), suggesting much better agreement between the techniques (ICC 0. 87; 95% confidence interval 0.86, 0.88). Among the physicians, agreement between DRE and TRUS was higher after training (ICC 0.64 to 0.96) than before training (ICC 0.02 to 0.49). CONCLUSIONS: Although the subjects examined before and after training differed, the agreement between TRUS and DRE prostate size estimates by the FPs appeared to be stronger after training with a three dimensional prostate model. This model may be a useful tool to assist in training FPs and medical students to measure prostate size on DRE. PMID- 10792083 TI - Clinical value of human glandular kallikrein 2 and free and total prostate specific antigen in serum from a population of men with prostate-specific antigen levels 3.0 ng/mL or greater. AB - OBJECTIVES: To investigate the clinical value of human glandular kallikrein 2 (hK2) compared with free (f) and total (t) prostate-specific antigen (PSA) in the early detection of prostate cancer (PCa). METHODS: In PCa screening conducted in 1995 to 1996 in Goteborg, Sweden, 5853 of 9811 randomly selected men (aged 50 to 66 years; median 61) accepted PSA testing; those with tPSA levels of 3. 0 ng/mL or greater were offered digital rectal examination, transrectal ultrasound, and sextant biopsies. Serum from 604 of 611 biopsied men (18% with positive digital rectal examinations, tPSA range 3.0 to 220 ng/mL, 144 men with PCa) was analyzed for hK2 (research assay) and tPSA and fPSA (Prostatus). Sera were stored at -20 degrees C for a maximum of 2 weeks for tPSA and fPSA and 3 years for hK2. RESULTS: hK2 levels and hK2 x tPSA/fPSA values were significantly elevated in men with PCa. Receiver operating characteristic data revealed that the area under the curve for hK2 x tPSA/fPSA was significantly greater than that for tPSA and greater, but not significantly greater, than that for percent fPSA. Also, the cancer-detecting sensitivity was significantly improved (P <0.05) using hK2 x tPSA/fPSA compared with tPSA and percent fPSA at specificity levels of 75% to 90%. At 75% specificity, a sensitivity of 74% was obtained compared with 64% or 54% using percent fPSA or tPSA; at 90% specificity, the corresponding sensitivity level was 55%, 41%, and 36%, respectively. CONCLUSIONS: Discrimination of men with and without PCa in a randomly selected population was improved by measuring hK2 in addition to tPSA and fPSA. PMID- 10792084 TI - Comparison of two investigative assays for the complexed prostate-specific antigen in total prostate-specific antigen between 4.1 and 10.0 ng/mL. AB - OBJECTIVES: To determine the ability of complexed prostate-specific antigen (cPSA) levels to diagnose prostate cancer. METHODS: Between September 1998 and March 1999, cPSA levels in 182 consecutive patients with an abnormal digital rectal examination (DRE) or a total PSA (tPSA; Tandem-R assay) level greater than 4.1 ng/mL were examined. Levels of cPSA were measured by the Markit-M PSA-ACT (alpha(1)-antichymotrypsin) assay (cPSA-MM) and Bayer Immuno 1 complexed PSA assay (cPSA-BI). Free PSA (fPSA) was measured by the Tandem-R free PSA assay. RESULTS: Of the 140 patients with tPSA between 4.1 and 10.0 ng/mL, 116 were histologically confirmed as having benign tissue; the remaining 24 were diagnosed with prostate cancer. To ensure a 92% sensitivity of cancer detection, a cutoff value for the tPSA, cPSA-MM, and cPSA-BI assays of 4.8 ng/mL, 2.7 ng/mL, and 4.6 ng/mL, respectively, was determined. The percentage of negative biopsies prevented at these cutoff (ie, specificity) values was 14%, 23%, and 24%. No significant differences among these three assays were found. At 92% sensitivity, the cutoff value for the fPSA/tPSA, fPSA/cPSA-MM, and fPSA/cPSA-BI ratios was 18%, 27%, and 18%, respectively. The specificity was 35%, 49%, and 51%. No significant differences were found among these three fPSA ratios. Significant differences were noted between tPSA and the fPSA/cPSA-MM ratio and between tPSA and the fPSA/cPSA-BI ratio. No differences were seen among the other PSA parameters. CONCLUSIONS: No difference in the ability of cPSA levels to distinguish prostate cancer and noncancer was observed between cPSA-MM and cPSA BI or between their fPSA ratios. Only the fPSA/cPSA-MM and fPSA/cPSA-BI ratios provided significantly enhanced diagnostic performance compared with tPSA. PMID- 10792085 TI - Correlation of prostate-specific antigen before prostate cancer detection and clinicopathologic features: evaluation of mass screening populations. AB - OBJECTIVES: Although prostate-specific antigen (PSA) has become the reference standard for prostate cancer diagnosis, few reports have examined the long-term changes in PSA values before the diagnosis of prostate cancer in a large number of subjects. We investigated serial PSA levels and related values before prostate cancer diagnosis in a mass screening population and analyzed the values in an attempt to discover some values useful in clinical diagnostic science. METHODS: We performed mass screening for prostate cancer in 9671 subjects from 1986 to 1998. The initial screening method was measurement of prostatic acid phosphatase from 1986 to 1991 and measurement of PSA from 1992 to 1998. As a result, 303 cases of prostate cancer were diagnosed. For all the cases diagnosed before 1991, we measured the serum PSA value in preserved frozen serum. RESULTS: The prostate cancer detection rate was 3.1% among all subjects observed during the 13-year period. By measurement of the PSA level using frozen serum during the pre-PSA era, we found that 62% of patients demonstrated a PSA abnormality for more than 1 year (average 2.8) before prostate cancer diagnosis. Prostate cancer that was diagnosed within 1 year after a PSA value became abnormal was not associated with bone metastasis. Concerning the relationship between PSA velocity (PSAV) and clinical stage, the proportion of Stage B cancer was 86% in the subjects whose PSAV level before diagnosis was 0.18 ng/mL/yr or less and it was only 29% in those with PSAV levels of 4.5 ng/mL/yr or more. Only 3 (3.5%) of 86 patients with prostate cancer with PSAV levels of 4.4 ng/mL/yr or less had bone metastasis, and 2 of those 3 patients had poorly differentiated adenocarcinoma. CONCLUSIONS: Although a total of 62% of patients had an abnormal PSA level more than 1 year before prostate cancer diagnosis, no patients with prostate cancer who were diagnosed within 1 year after the PSA level became abnormal had bone metastasis. Among patients who have undergone mass screening twice or more, a clinically useful indicator of the lack of bone metastasis would be a period between the detection of PSA levels of 4.1 ng/mL or more but not more than 10 ng/mL and prostate cancer diagnosis of less than 1 year and a diagnosis of well or moderately differentiated adenocarcinoma or a PSAV of 4.4 ng/mL/yr or less and a cancer diagnosis of well or moderately differentiated adenocarcinoma. PMID- 10792086 TI - Total PSA, free PSA/total PSA ratio, and molecular PSA detection in prostate cancer: which is clinically effective and when? AB - OBJECTIVES: To ascertain when the serum determination of the free prostate specific antigen (PSA)/total PSA (fPSA/tPSA) ratio is clinically useful, and whether the identification of PSA or prostate-specific membrane antigen (PSM) mRNA in circulating cells has diagnostic advantages over the determination of their protein product. METHODS: fPSA, tPSA, and the fPSA/tPSA ratio were determined in the sera of 50 men with benign nonprostatic urologic diseases (EPD), 112 patients with prostate cancer (PCa), and 218 with benign prostatic hyperplasia (BPH). mRNA was extracted from the circulating mononuclear cells of 13 EPD samples, 25 PCa samples, and 38 BPH samples. PSA and PSM mRNA signals were identified in these samples by means of reverse transcriptase-polymerase chain reaction. RESULTS: Overall, at a fixed specificity of 95%, the sensitivity of tPSA was 19% and that of the fPSA/tPSA ratio was 40% in distinguishing PCa from BPH. The fPSA/tPSA ratio allowed the discrimination of PCa from BPH with satisfactory sensitivity and specificity when considering patients less than 60 years of age (100% and 95%, respectively). PSA and PSM mRNA were positive in 1 and 7 of 13 EPD samples, 6 and 13 of 25 PCa samples, and 6 and 17 of 38 BPH samples. The Gleason score did not correlate with tPSA, the fPSA/tPSA ratio, PSA mRNA, or PSM mRNA. CONCLUSIONS: The serum determination of the fPSA/tPSA ratio is an excellent index of PCa for subjects younger than 60 years of age; the clinical utility of PSA mRNA identification in circulating cells needs to be validated by large follow-up studies, and the analysis of PSM mRNA seems to be of no clinical interest. PMID- 10792087 TI - African-American men and intention to adhere to recommended follow-up for an abnormal prostate cancer early detection examination result. AB - OBJECTIVES: To assess the intention of African-American men to have the recommended follow-up in the event of an abnormal prostate cancer early detection examination and to identify the variables that help to explain adherence intention. METHODS: In the spring of 1995, we selected a random sample of 548 African-American men who were patients at the University of Chicago Health Service. The sample included men who were 40 to 70 years of age, did not have a personal history of prostate cancer, and had a working telephone number. A total of 413 men who completed the telephone survey received an invitation to consider undergoing a prostate cancer early detection examination. The survey provided data on personal background characteristics, knowledge, attitudes, and beliefs related to prostate cancer and early detection. Respondents were asked whether they would choose to have the recommended follow-up in the event of an abnormal early detection examination result. Univariate and multivariate analyses of intention to have follow-up were performed. RESULTS: An intention to have the recommended follow-up was reported by 77% of the survey respondents. The results of multivariate analyses revealed that the intention to have the follow-up was positively associated with education beyond high school (odds ratio [OR] 1.9); perceived self-efficacy related to prostate cancer screening (OR 2.1); the belief that prostate cancer can be cured (OR 3.3); the belief that prostate cancer screening should be done in the absence of prostate problems (OR 2.3); and physician support for prostate cancer screening (OR 2.1). CONCLUSIONS: African American men who have a high school education or less may be at risk of nonadherence to recommended follow-up. Adherence also may be low among men who do not have favorable views of early detection or do not perceive strong physician support for early detection. Research is needed to determine whether intention and other factors predict actual adherence to follow-up in this population group. PMID- 10792089 TI - Editorial comment PMID- 10792088 TI - Urolume stent placement for the treatment of postbrachytherapy bladder outlet obstruction. AB - OBJECTIVES: Transurethral resection (TURP) or incision of the prostate is generally not effective for treating bladder outlet obstruction after transperineal brachytherapy for prostate cancer. Furthermore, TURP could compromise full-dose effective radiation delivery to the prostate. We analyzed the efficacy of the UroLume stent in treating the urinary outflow obstruction in such patients. METHODS: Five patients who had undergone brachytherapy (3 with (192)Ir high-dose radiation and 2 with (125)I) subsequently developed one or more episodes of urinary retention 2 weeks to 4 years after treatment. The patients failed or could not tolerate alpha-blockers or clean intermittent catheterization. Three patients subsequently underwent urethral dilation/optical internal urethrotomy for strictures, and 1 patient underwent suprapubic tube placement. Following the failure of these interventions, each of these patients had a UroLume stent placement. A single UroLume stent (2 cm in 3 patients and 2.5 cm in 2 patients) was placed under local/spinal anesthesia. RESULTS: All patients were able to void spontaneously immediately after stent placement. Of the patients with previous urethral strictures, 1 remained continent and 1 had persistent incontinence. Neither of the patients with early postbrachytherapy retention developed incontinence after stent placement. The main complaints following stent placement were referred pain to the head of the penis and dysuria. Stent-related symptoms necessitated stent removal in 2 of 5 patients, 4 to 6 weeks after placement. CONCLUSIONS: The UroLume stent can be used as an alternative form of therapy for managing postbrachytherapy bladder outlet obstruction. The treatment is easily reversible by removing the stent when obstruction resolves. PMID- 10792090 TI - Failure patterns and hazard rates for failure suggest the cure of prostate cancer by external beam radiation. AB - OBJECTIVES: To present patterns of failure and hazard rates for failure that support the concept of cure for patients with prostate cancer treated with external beam radiation (RT). METHODS: Two patient groups are reported: 408 patients treated with RT alone and 63 patients treated with RT and short-term androgen deprivation (RT+AD). All patients were treated between March 1987 and March 1995 and had at least 4 years of prostate-specific antigen (PSA) follow-up. The median follow-up was 69 months for the RT alone group and 60 months for the RT+AD group. For each treatment group, biochemical control and hazard functions were estimated using the ASTRO consensus definition of failure and the life table method. RESULTS: The 5 and 8-year biochemical control estimates were 60% and 59% for the RT alone group, respectively, with only two failures occurring after 5 years (1% of the total failures observed). Hazard function estimates indicated a maximum risk of failure at 12 to 36 months, tapering to a low rate at 4 years, with no failures observed after 6 years. The differences in the patterns of failure by PSA level revealed a maximum risk of failure at 12 to 24 months (median 28) for a pretreatment PSA level of less than 10 ng/mL, 12 to 36 months (median 25) for a pretreatment PSA level of 10 to 19.9 ng/mL, and 12 to 36 months (median 22) for a pretreatment PSA level of 20 ng/mL or greater. The latter group reached low levels of risk at 6 years in contrast to 4 years for the patients presenting with pretreatment PSA levels of less than 20 ng/mL. Similar patterns were observed when stratifying by stage and Gleason score: patients with a worse prognosis had the highest risk of failure earlier and achieved a low risk of failure later than patients with a more favorable prognosis. The patients in the RT+AD group had a different pattern of risk of failure, with the highest risk immediately after treatment, declining to a low risk of failure at 48 months. CONCLUSIONS: Patients treated with RT alone or RT+AD had little risk of failure after 4 to 6 years. Patients with a favorable prognosis achieved a low risk of failure sooner than high-risk patients when treated with RT alone. These results are consistent with the cure of prostate cancer by RT alone or RT+AD. PMID- 10792091 TI - Explaining the difference in prostate cancer mortality rates between white and black men in the United States. AB - OBJECTIVES: Black men have a twofold increased risk of dying of prostate cancer compared to white men. We assessed the extent to which differences in stage and grade at diagnosis, age, multiple primary cancers, and the first course of cancer directed therapy influence the likelihood of prostate cancer death among diagnosed cases. METHODS: Incidence-based mortality (IBM) is a method that involves tracking disease cases from diagnosis to death, such that the mortality event is linked to factors identified with the disease at the time of diagnosis (eg, tumor stage and grade). We applied the IBM method to prostate cancer data from the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute. RESULTS: Of men diagnosed with prostate cancer, blacks have a significantly greater chance of dying from the disease than do whites. However, once adjustment has been made for stage and grade at diagnosis, age, number of primary cancers, and initial treatment, no difference in the likelihood of prostate cancer mortality between the two races was found. Differences in stage and grade at diagnosis, age, number of primary cancers, and initial treatment each contributed significantly to the greater chance of dying of prostate cancer for black men than for white men. CONCLUSIONS: The results of IBM analysis suggest that black men do not have a statistically greater chance of prostate cancer mortality than white men after adjusting for differences in stage and grade, age, number of primary cancers, and treatment. Later stage at diagnosis is the primary reason for the higher likelihood of prostate cancer mortality among black men compared to white men. PMID- 10792092 TI - Quality-of-life comparison of radical prostatectomy and interstitial brachytherapy in the treatment of clinically localized prostate cancer. AB - OBJECTIVES: Interstitial brachytherapy (BT) is increasingly utilized as a curative treatment for localized prostate cancer because it is perceived as less morbid than surgical alternatives. However, to date no studies have directly compared the quality of life and symptoms of patients with localized prostate cancer treated with curative intent by radical prostatectomy with those treated by either BT alone or BT combined with external beam radiation. METHODS: On June 1, 1998, 242 men with clinically localized Stage T1c to T3 adenocarcinoma of the prostate, treated at our institution with curative intent from January 1, 1997 to June 1, 1998, were mailed a questionnaire. Cross-sectional analysis of returned questionnaires was carried out. Patients were treated with either radical prostatectomy (RP), palladium-103 (Pd(103)) brachytherapy (115 Gy) monotherapy (BTM), or Pd(103) combined brachytherapy (90 Gy) and external beam radiation (40 to 45 Gy) (BTC). The primary outcome measures were the Functional Assessment of Cancer Therapy scale (FACT-G), American Urological Association (AUA)/international prostate symptom score (IPSS), "Urinary Function Questionnaire for Men after Radical Prostatectomy," and Brief Sexual Function Inventory. RESULTS: Data from 138 patients were included in the analysis; 27 had RP, 70 had BTM, and 41 had BTC. Total FACT-G and personal well-being scores were significantly lower in the BTC group. Brachytherapy monotherapy and RP had similar scores on the FACT-G, with surgical patients having the lowest IPSS scores. Correlations were noted between total FACT-G and urinary symptom score, degree of sexual function, frequency of diarrhea, and frequency of hot flashes. Bothersomeness of urinary function correlated with the degree of urinary control. The radical prostatectomy and BTM groups had improvement in quality of life, voiding, diarrhea, and sexual function with time, whereas the BTC group experienced a decline. CONCLUSIONS: Patients treated with BTC had an overall lower quality of life compared with those treated by RP and BTM, and RP patients reported fewer irritative or obstructive voiding complaints. Although the consistency and magnitude of these trends require further study, our data suggest that RP remains a well-tolerated and accepted option. PMID- 10792093 TI - Microvessel density, p53, retinoblastoma, and chromogranin A immunohistochemistry as predictors of disease-specific survival following radical prostatectomy for carcinoma of the prostate. AB - OBJECTIVES: Angiogenesis has been shown to be related to p53 and retinoblastoma gene function as well as to neuroendocrine differentiation (as measured by chromogranin A staining) in prostate tumors. Studies have indicated that immunohistochemical assessment of p53, retinoblastoma, and chromogranin A in prostate cancers treated by radical prostatectomy can be useful in predicting disease-specific survival, whereas the degree of microvessel density (MVD), a measure of angiogenesis, correlates with disease recurrence. The ability of MVD, however, to predict disease-specific survival either alone or in conjunction with other prognostic factors has not yet been evaluated. The purpose of our study was to determine the relative importance of p53, retinoblastoma, and chromogranin A as well as MVD in the prediction of disease-specific survival following radical prostatectomy in conjunction with classical pathologic assessment. METHODS: From 1970 to 1984, radical prostatectomy was performed on 75 patients with clinical Stage A2 to B2 adenocarcinoma of the prostate. No neoadjuvant or adjuvant treatments were given, and patients were followed until death. Prostatectomy specimens were examined to evaluate conventional pathologic parameters. In addition, the tissue was immunohistochemically stained for p53, retinoblastoma, chromogranin A, and endothelial cells. A previously described computerized imaging system analyzed the microvessels and computed both "optimized" and "area weighted" MVD scores. Proportional hazard models were used to investigate the simultaneous association of these variables with disease-specific survival. RESULTS: Of the 75 patients, 4 had follow-up of less than 3 months, and 29 patients had inadequate tissue for analysis of all immunohistochemical markers. The analyzed subset of 42 patients was found to be representative of the cohort of 71 patients. Multivariate analysis revealed that p53 and retinoblastoma have the greatest prognostic importance regarding disease-specific survival. Chromogranin A and optimized or area-weighted MVD scores were of no additional value when p53 and retinoblastoma were assessed. CONCLUSIONS: Microvessel density, as a determinant of angiogenesis and chromogranin A, does not seem to add significantly to the prognostic disease-specific survival information provided by conventional pathology combined with p53 and retinoblastoma assessment. PMID- 10792094 TI - Comparison of results and complications of high ligation surgery and microsurgical high inguinal varicocelectomy in the treatment of varicocele. AB - OBJECTIVES: To prospectively compare sperm parameters, pregnancy and recurrence rates, and complications after randomized high ligation surgery versus microsurgical high inguinal varicocelectomy (MHIV). METHODS: Varicocele was diagnosed by physical examination and color Doppler ultrasound in 468 patients who underwent one of two procedures: high ligation surgery (n = 232) or MHIV (n = 236). The high ligation surgery was left unilateral in 142 and bilateral in 90. The MHIV was left unilateral in 128 and bilateral in 108. The patients were postoperatively evaluated by spermiograms and physical examination. The pregnancy rate was monitored for 2 years. RESULTS: One year after surgery, 34.05% in the high ligation group and 46.61% in the MHIV group had a more than 50% increase in their total motile sperm count (P = 0.000). The increase in sperm count was not statistically different between the two groups (P = 0.1), but the difference in the increase in sperm motility in the MHIV group was statistically significant (P = 0.000). Pregnancy rates at the end of 2 years reached 33.57% in the high ligation group and 42.85% in the MHIV group, not a statistically significant difference (P = 0.0571). The postoperative recurrence as detected by physical examination was markedly different between the two techniques. The recurrence rate was 15.51% in the high ligation group and 2.11% in the MHIV group (P = 0.000). Also, the incidence of postoperative hydrocele was significantly different between the two groups (9.09% in the high ligation group and 0.69% in the MHIV group; P = 0.000). CONCLUSIONS: MHIV has lower recurrence and hydrocele rates, a higher increase in sperm motility, and results in higher pregnancy rates. Therefore, it should be the preferred technique for varicocelectomy. PMID- 10792095 TI - Serum dehydroepiandrosterone sulfate concentrations in men with erectile dysfunction. AB - OBJECTIVES: In 1994, the Massachusetts Male Aging Study presented the finding of an inverse correlation of the serum levels of dehydroepiandrosterone sulfate (DHEAS) and the incidence of erectile dysfunction (ED). Prompted by the positive results of a pilot study on the treatment of ED with dehydroepiandrosterone (DHEA), we performed a detailed investigation on the serum DHEAS levels in men with ED according to age category. METHODS: Inclusion criteria included a history of ED for more than 6 months, a body mass index less than 30, and a state of good general health. Serum DHEAS concentrations were determined in 309 patients with ED and 133 healthy volunteers. All participants were carefully screened to assess medical factors known or suspected to alter endocrine function. Questions 3 and 4 of the International Index of Erectile Function were used to evaluate erectile function. RESULTS: The mean serum levels of DHEAS in patients with ED were lower than in healthy volunteers until 60 years of age. The shape of the curve of the patients with ED indicated a quadratic decrease of DHEAS with age in contrast to a more linear decrease of DHEAS with age in the control group. CONCLUSIONS: Our results suggest that until the age of 60 years, the mean serum level of DHEAS is lower in patients with ED than in healthy volunteers. PMID- 10792097 TI - Editorial comment PMID- 10792096 TI - Failure of subureteral bovine collagen injection for the endoscopic treatment of primary vesicoureteral reflux in long-term follow-up. AB - OBJECTIVES: To evaluate the long-term efficacy of subureteral glutaraldehyde cross-linked collagen injection (GAX 35) for endoscopic treatment of primary vesicoureteral reflux (VUR). METHODS: We prospectively studied 36 patients (58 ureteral renal units), 30 girls and 6 boys with a median age of 6 years (range 2 months to 18 years). All patients had primary VUR and were treated with a single subureteral collagen injection (GAX 35). The patients were followed up by voiding cystography. RESULTS: According to the International Reflux Study Classification, we found the following reflux grades preoperatively: grade I, 2 ureteral units; grade II, 21 units; grade III, 28 units; grade IV, 4 units, and grade V, 3 units. All patients were treated with subureteral bovine collagen injection (GAX 35, mean volume 1.7 mL, range 0.7 to 3.5). All but 3 cases of reflux resolved initially. The mean follow-up was 13 months (range 1 to 108). After 37 months of follow-up, only 5 (9%) of 57 treated units remained reflux free. One unit was followed up for 17 months and also remained reflux free. CONCLUSIONS: These data suggest that a single endoscopic subureteral collagen injection is not effective in the long-term follow-up of patients with primary VUR. In the future, it will be important to determine whether the new, currently used, and soon be approved bulking agents show better long-term clinical results to prevent VUR recurrence than bovine collagen does. PMID- 10792098 TI - Penile reconstruction combined with bladder closure in the management of classic bladder exstrophy: illustration of technique. AB - OBJECTIVES: Interest has increased in combining procedures during reconstruction of bladder exstrophy in an effort to reduce the number of procedures required for reconstruction and to improve results. This study illustrates our technique of reconstruction and summarizes our current experience with the combination of epispadias repair and bladder closure during initial reconstruction or following prior failed bladder closure. METHODS: Twenty-four boys with classic bladder exstrophy underwent combined bladder closure and epispadias repair. The mean patient age was 20 months, and 18 boys had a prior failed closure. Osteotomies were performed in all patients. RESULTS: No instances of bladder prolapse or dehiscence were noted on follow-up. Urethrocutaneous fistula developed in 7 patients. Eventual bladder neck reconstruction has been performed in 11 boys (6 boys are dry day and night, 3 are dry during the day with occasional wet episodes at night, 2 have required follow-up bladder augmentation and continent diversion for persistent incontinence), and 1 boy had augmentation at the same time as bladder neck reconstruction. Twelve boys are awaiting adequate capacity for bladder neck reconstruction, and 1 is awaiting bladder augmentation and continent stoma construction. CONCLUSIONS: Epispadias closure can be safely combined with bladder closure in select patients with classic bladder exstrophy. Complication rates and cosmesis approximate that achieved with staging the two procedures. This achievement represents strict patient selection and attentive follow-up. PMID- 10792099 TI - Abdominoscrotal hydrocele: a reliable surgical technique. AB - Abdominoscrotal hydrocele (ASH) consists of two large sacs, both abdominal and scrotal, connecting with the inguinal channel. The diagnosis is made only by ultrasound scan. Surgical treatment is mandatory since no spontaneous resolution has been reported. A new surgical procedure used successfully to treat 11 patients with ASH is described. PMID- 10792100 TI - 8p22 loss concurrent with 8c gain is associated with poor outcome in prostate cancer. AB - OBJECTIVES: A critical issue in the management of prostate cancer is the ability to distinguish patients at risk of disease recurrence. The aim of this study was to determine whether specific physical alterations of chromosome 8 may be associated with disease recurrence and poor outcome using postoperative prostate specific antigen (PSA) values as surrogate end points. METHODS: To test this hypothesis, we examined paired normal and tumor radical prostatectomy tissues from 25 patients with prostate cancer for chromosome 8 alterations using dual fluorescence in situ hybridization with a fluorescein-labeled 8p22-specific (8p) cosmid probe and a rhodamine-labeled 8-centromere-specific (8c) probe. The probes were enumerated in 200 nuclei per tissue. RESULTS: Of the 25 tumors examined, 22 demonstrated distinct classes of genetic alterations, or nuclear types, including disomy for 8p and 8c (1 tumor), loss of 8p and disomy for 8c (10 tumors), or loss of 8p concurrent with gain of 8c (11 tumors). The presence of even a small population of tumor nuclei characterized by the loss of 8p concurrent with the gain of 8c was correlated with poor tumor grade (P = 0.009), preoperative PSA values 11 ng/mL or higher (P = 0.022), high tumor stage (P = 0.086), and detectable, rising postoperative PSA values (P = 0.086). These observations are consistent with the hypothesis that a gain of chromosome 8 is associated with poor outcome in prostate cancer. CONCLUSIONS: 8p loss concurrent with 8c gain may successfully predict disease recurrence and poor clinical outcome before the observation of detectable postoperative PSA values in patients with prostate cancer. PMID- 10792101 TI - Optical coherence tomography as a method for identifying benign and malignant microscopic structures in the prostate gland. AB - OBJECTIVES: Optical coherence tomography (OCT) is a new optical imaging technique capable of providing cross-sectional imaging of tissue microstructure in vivo and in real time. OCT was used in the setting of the human prostate ex vivo, and the images acquired were compared with those obtained using standard histopathologic methods. METHODS: Multiple samples (3 to 6) were obtained from the radical prostatectomy specimens of 7 men with clinically localized (T1c-2, N0, M0) adenocarcinoma of the prostate. These specimens were 1 cm in length and 1 mm x 1 mm in rectangular cross section. Specimens were first imaged using OCT and then embedded and stained in preparation for histopathologic evaluation. Co registration of the images obtained using OCT and standard histopathologic evaluation provided the basis for comparison. RESULTS: Structural architecture on the order of 50 to 150 microm within benign glandular epithelium, fibroadipose tissue, and malignant glandular epithelium could be resolved to a depth of approximately 0.5 mm using OCT. CONCLUSIONS: Microscopic resolution is possible in human prostatic tissue using OCT. Further studies using this technique to improve the detection and staging of adenocarcinoma of the prostate are ongoing. PMID- 10792102 TI - Urethral atresia in a neonate with alveolar capillary dysplasia and pulmonary venous misalignment. AB - Urethral atresia and alveolar capillary dysplasia (ACD) are rare congenital malformations. Urethral atresia is associated with severe pulmonary hypoplasia secondary to oligohydramnios. ACD is associated with pulmonary venous misalignment, results in severe pulmonary hypertension, and is uniformly fatal. We present a case of urethral atresia with successful, early placement of vesicoamniotic shunting, with resolution of the oligohydramnios, in which the neonate rapidly progressed to respiratory failure and death. Postmortem examination confirmed urethral atresia and diagnosed ACD. Given the surprisingly high mortality rate after vesicoamniotic shunting in patients with urethral atresia, we question whether there might be a possible link to ACD. PMID- 10792103 TI - Megacystis microcolon intestinal hypoperistalsis syndrome: bladder distension and pyelectasis in the fetus without anatomic outflow obstruction. AB - We report a case of a 46,XY male infant with a history of normal amniotic fluid levels who was delivered by elective cesarean section at 38.5 weeks' gestation because of progressive bladder distension, hydroureteronephrosis, and what was thought to be a dilated posterior urethra. The child died at 19 days of age of cardiovascular complications. The autopsy revealed megacystis, bilateral megaureters and pyelocaliectasis, congenital absence of ganglion cells in the bladder wall, renal dysplasia, and a microcolon. No dilation or anatomic obstruction of the posterior urethra was found. These findings strongly suggest the diagnosis of megacystis microcolon intestinal hypoperistalsis syndrome. We discuss the ultrasound findings of in utero bladder distension with hydronephrosis and one of its rare etiologies. PMID- 10792104 TI - Successful in utero endoscopic ablation of posterior urethral valves: a new dimension in fetal urology. AB - Fetal lower urinary tract obstructive uropathy, when associated with oligohydramnios, is usually associated with a poor outcome. We present a case of successful in utero endoscopic ablation of posterior urethral valves in which the infant survived the neonatal period without evidence of renal dysplasia. The role, indications, and potential benefits of this novel technique are discussed. PMID- 10792105 TI - Complications of intrauterine intervention for treatment of fetal obstructive uropathy. AB - The intrauterine surgical placement of vesicoamniotic shunts in the treatment of fetal obstructive uropathy associated with prune-belly syndrome to avoid such complications as renal damage and oligohydramnios remains controversial. We present a case of an infant born with prune-belly syndrome at 33 weeks and 5 days of estimated gestational age to a mother of two by vaginal delivery after a pregnancy complicated by fetal obstructive uropathy with attempted intrauterine intervention. After sonographic and laboratory diagnostic and prognostic evaluations, an intrauterine procedure was performed in which a vesicoamniotic shunt was placed under ultrasound guidance. Complications included dislodgment of the initial shunt, with a failed subsequent attempt at placement, oligohydramnios, preterm labor and delivery, and traumatic gastroschisis through the surgical abdominal wall defect. His hospital stay was further complicated by chronic renal insufficiency, prematurity, respiratory distress, bowel malrotation, an episode of gram-negative sepsis with Enterobacter cloacae, signs of liver failure, an exploratory laparotomy for severe enterocolitis, and orchiopexy for bilateral undescended testes. At present, it is unclear whether vesicoamniotic shunt placement can provide any significant improvement in the morbidity or mortality for patients with prune-belly syndrome. A large, prospective, randomized trial is needed to determine its efficacy. PMID- 10792106 TI - Antenatally detected proximal ureteral diverticulum. AB - We present what is to our knowledge the first antenatal detection of a proximal ureteral diverticulum, an extremely rare urologic abnormality, that was successfully reconstructed in the early postnatal period and should henceforth be included in the differential diagnosis of the antenatally detected cystic mass. PMID- 10792107 TI - Dismembered pyeloplasty followed by metachronous ureteropelvic junction obstruction in the contralateral kidney. AB - Hydronephrosis due to congenital ureteropelvic junction obstruction (UPJO) is commonly diagnosed by antenatal sonography. We report the case of an infant who developed new-onset hydronephrosis in the right kidney following uneventful left sided pyeloplasty for a congenital UPJO. The furosemide minus 15-minute diuretic renogram (F-15 DR) was used to confirm obstruction in the affected renal unit when standard diuretic renography was equivocal. The indications for surgery or observation as well as the role of the F-15 DR are discussed. PMID- 10792108 TI - Cloacal outlet obstruction with an ectopic ureter. AB - Cloacal malformation occurs in approximately 1 in 50,000 live female births. Prenatal ultrasound may lead to the diagnosis in selected cases. We report an unusual case of prenatally detected single-system hydronephrosis with a nonvisible bladder and worsening oligohydramnios. Labor was induced at 35 weeks' estimated gestational age. On physical examination, a single perineal opening was noted consistent with cloaca. Endoscopy revealed an obstructed ectopic ureter at the level of the sphincter, an undeveloped bladder and vagina, and a fistula to the rectum. A low loop cutaneous ureterostomy and right upper quadrant loop colostomy were performed. The absence of a typical fluid-filled pelvic structure may confound the prenatal diagnosis of cloaca. PMID- 10792109 TI - Intrarenal cystic mass with pelviureteral junction obstruction. AB - An abdominal mass in the first year after birth most commonly originates from the kidney. Renal masses in this age group are often cystic and may be associated with other abnormalities. We describe an unusual benign unilateral cystic lesion and coexisting pelviureteral junction obstruction in a newborn male with an otherwise morphologically and functionally normal urinary tract. A near normal kidney resulted from subsequent resection of the cyst wall combined with pyeloplasty. PMID- 10792110 TI - Hyperreninemia and congenital mesoblastic nephroma: case report and review of the literature. AB - A 32-week estimated gestational age female infant had elevated systolic blood pressure at birth and a palpable left abdominal mass. Radiologic evaluation revealed a left upper pole renal mass. The renin level was significantly elevated. Left nephrectomy confirmed a diagnosis of congenital mesoblastic nephroma. The postoperative renin level normalized along with the blood pressure. Histologic analysis identified renin production from either the mesoblastic nephroma or secondarily from compression or ischemia. Mesoblastic nephroma should be considered as part of the differential diagnosis for hypertension in the young. The renin concentration and a renal ultrasound scan should be obtained as part of the evaluation. PMID- 10792111 TI - Review article: the clinical role of anti-TNFalpha antibody treatment in Crohn's disease. AB - The recent licensing of anti-TNFalpha antibody treatment offers the potential to radically alter the course of severe Crohn's disease using genetically-engineered drugs directed against a specific inflammatory mediator. Controlled randomized trials have demonstrated clinical benefit associated with tissue healing in patients with active intestinal disease and fistulae, often when conventional therapies were unsuccessful. This therapy is expensive, however, and long-term efficacy and safety data are still awaited. This review considers the nature of this therapy and the current evidence for its clinical benefit and adverse effects. The treatment is also considered in the context of available immunosuppressive agents, with suggestions about its practical application. PMID- 10792112 TI - Review article: pharmacological treatment of the hepatorenal syndrome in cirrhotic patients. AB - Renal failure is common in patients who are dying from end-stage cirrhosis, developing in 40-80% of all patients. Where there is no anatomical or pathological cause for the renal failure, it is termed the hepatorenal syndrome. When the hepatorenal syndrome develops, it will only recover when there is some degree of improvement in liver function. Thus for most patients this will occur only after liver transplantation, although the transplantation mortality is increased in this group. Hepatorenal syndrome is a common complication of alcoholic hepatitis, and this group is unusual in that with time and abstinence, significant recovery of liver function may occur. There is therefore a need for supportive therapy to allow time for some recovery of liver function in patients with alcoholic hepatitis and hepatorenal syndrome. Similarly, patients may need support whilst waiting for liver transplantation. This article reviews the pathophysiology and treatment of hepatorenal syndrome. PMID- 10792113 TI - Review article: new insights into the pathogenesis of radiation-induced intestinal dysfunction. AB - Exposure of the abdomino-pelvic region to ionizing radiation, such as that received during radiotherapy, is associated with the development of a number of untoward symptoms which may limit the course of therapy or which may involve serious chronic intestinal disease. While the mucosal dysfunction surrounding acute radiation enteritis is generally ascribed to the effects of ionizing radiation on the cell cycle of epithelial stem cells of the intestinal crypts and subsequent epithelial loss, recent evidence suggests that other, earlier events also play a role. The severity of these early events may determine the incidence and severity of chronic enteritis. The mechanism for this is unclear, but may relate to radiation-induced compromise of host defence responses to luminal pathogens or antigens. This review will address the current state of knowledge of the pathogenesis of radiation-induced intestinal dysfunction, focusing on events which occur in the mucosa, and will discuss what the future may hold with respect to the treatment of radiation-associated diseases of the intestinal tract. PMID- 10792114 TI - Low-dose misoprostol for the prevention of low-dose aspirin-induced gastroduodenal injury. AB - INTRODUCTION: The harmful effects of non-steroidal anti-inflammatory drugs (NSAIDs) on the gastric mucosa and the prophylactic effects of misoprostol are both dose-dependent. AIM: To investigate whether a low-dose of misoprostol is sufficient to prevent gastric mucosal injury caused by low-dose aspirin. METHODS: We conducted a double-blind placebo controlled parallel group endoscopic study in 32 evaluable volunteers. The main outcome measure was erosive injury (ulcers and superficial erosions) in the gastric mucosa over 28 days. RESULTS: Most subjects developed erosions on aspirin 300 mg daily. This was significantly reduced by misoprostol 100 microg daily. (Odds ratio 0.18, 95% CI: 0.07-0.48). There were no drug-related or gastrointestinal adverse events in subjects receiving misoprostol. CONCLUSION: Misoprostol 100 microg daily can prevent low-dose aspirin induced gastric mucosal injury without causing identifiable adverse effects. PMID- 10792116 TI - Dose-dependent effects of ketoprofen on the human gastric mucosa in comparison with ibuprofen. AB - BACKGROUND: As non-steroidal anti-inflammatory drugs (NSAIDs) become available for over-the-counter use, it is important to define doses that would not cause undue gastroduodenal damage during the short periods for which self-medication with NSAIDs is licensed. AIM: To establish what dose of ketoprofen most closely resembles the maximum dose of ibuprofen (400 mg t.d.s.) licensed for self medication. METHODS: We studied healthy volunteers in a double-blind double-dummy randomized crossover study. Each subject took, over four separate 10-day dosing periods, ibuprofen 400 mg t.d.s., ketoprofen 12.5 mg t.d.s., ketoprofen 25 mg t.d.s. or ketoprofen 50 mg t.d.s. Mucosal injury was assessed by endoscopy at baseline and on the 3rd and 10th day of each dosing period. Ex vivo gastric mucosal prostaglandin (PG) E2 evoked by vortex mixing was measured by radioimmunoassay. Serum thromboxane was also measured by radioimmunoassay. RESULTS: Ketoprofen 50 mg t.d.s. suppressed prostaglandin synthesis to a significantly greater extent than ibuprofen and caused significantly more gastroduodenal injury. The profile of prostaglandin synthesis and injury on ketoprofen 12.5 mg t.d.s. most closely resembled that of ibuprofen 400 mg t.d.s. CONCLUSIONS: Ketoprofen 12.5 mg t.d.s. is an appropriate dose for self medication, which is likely to be similar to ibuprofen 400 mg t. d.s. in its effects on the stomach and duodenum. PMID- 10792115 TI - The effects of omeprazole on healing and appearance of small gastric and duodenal lesions during dosing with diclofenac in healthy subjects. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are associated with gastrointestinal mucosal damage. Omeprazole prevents the formation, and accelerates the healing, of NSAID-induced ulcers. AIM: To test whether omeprazole accelerates healing of standardized gastroduodenal lesions in the presence of diclofenac. METHODS: In a double-blind, double-dummy, placebo-controlled, crossover study, 12 healthy volunteers received consecutive, 2-week courses of omeprazole (40 mg o.d.) and placebo, in random order, with an intervening, 4-week washout period; diclofenac (50 mg t.d.s.), was given for the second week of each course. Five endoscopies were performed, one at the outset and the others before and after each course of diclofenac. Biopsies were taken from the endoscopically normal mucosa of the corpus, antrum and duodenum and also from any new mucosal lesion that developed after diclofenac. The sites of biopsies taken before each course of diclofenac were evaluated endoscopically after each course to assess the extent of healing according to a predetermined healing score scale. RESULTS: The healing scores observed after administration of placebo/diclofenac (median=0; range 0-6) and after omeprazole/diclofenac (median=0; range 0-6; P=0.17) did not differ. Small gastroduodenal lesions developed de novo in six subjects during placebo/diclofenac and in seven during omeprazole/diclofenac. Focal chemical gastropathy was observed only in close proximity to macroscopic lesions. CONCLUSIONS: In healthy subjects, omeprazole does not accelerate the healing of pre-existing mucosal lesions or prevent the development of small diclofenac induced mucosal lesions. PMID- 10792117 TI - Rifaximin versus chlortetracycline in the short-term treatment of small intestinal bacterial overgrowth. AB - BACKGROUND: Bacterial overgrowth of the small intestine is a condition characterized by nutrient malabsorption due to an excessive number of bacteria in the lumen of the small intestine. Current treatment is based on empirical courses of broad spectrum antibiotics; few controlled data, with respect to the duration and choice of antibiotic drug, exist at present. The recent availability of rifaximin, a non-absorbable rifamycin derivative, highly effective against anaerobic bacteria, prompted us to carry out a randomized, double-blind controlled trial in order to compare its efficacy and tolerability to those of tetracycline, currently considered the first-choice drug. METHODS: In 21 patients affected by small intestinal bacterial overgrowth, fasting, peak and total H2 excretion after ingestion of 50 g glucose and severity of symptoms were evaluated before and after a 7-day course of rifaximin, 1200 mg/day (400 mg t.d.s.), or chlortetracycline, 1 g/day (333 mg t.d.s. ). RESULTS: Fasting, peak and total H2 excretion decreased significantly in the group of patients treated with rifaximin whereas chlortetracycline did not modify these parameters. The H2 breath test normalized in 70% of patients after rifaximin and in 27% of patients after chlortetracycline. The improvement in symptoms was significantly higher in patients treated with rifaximin. CONCLUSIONS: Rifaximin is a promising, easily handled and safe drug for the short-term treatment of small intestinal bacterial overgrowth. PMID- 10792119 TI - Comparative effects of levosulpiride and cisapride on gastric emptying and symptoms in patients with functional dyspepsia and gastroparesis. AB - BACKGROUND: The efficacy of several prokinetic drugs on dyspeptic symptoms and on gastric emptying rates are well-established in patients with functional dyspepsia, but formal studies comparing different prokinetic drugs are lacking. AIM: To compare the effects of chronic oral administration of cisapride and levosulpiride in patients with functional dyspepsia and delayed gastric emptying. METHODS: In a double-blind crossover comparison, the effects of a 4-week administration of levosulpiride (25 mg t.d.s.) and cisapride (10 mg t.d.s.) on the gastric emptying rate and on symptoms were evaluated in 30 dyspeptic patients with functional gastroparesis. At the beginning of the study and after levosulpiride or cisapride treatment, the gastric emptying time of a standard meal was measured by 13C-octanoic acid breath test. Gastrointestinal symptom scores were also evaluated. RESULTS: The efficacy of levosulpiride was similar to that of cisapride in significantly shortening (P < 0.001) the t1/2 of gastric emptying. No significant differences were observed between the two treatments with regards to improvements in total symptom scores. However, levosulpiride was significantly more effective (P < 0.01) than cisapride in improving the impact of symptoms on the patients' every-day activities and in improving individual symptoms such as nausea, vomiting and early postprandial satiety. CONCLUSION: The efficacy of levosulpiride and cisapride in reducing gastric emptying times with no relevant side-effects is similar. The impact of symptoms on patients' everyday activities and the improvement of some symptoms such as nausea, vomiting and early satiety was more evident with levosulpiride than cisapride. PMID- 10792118 TI - Treatment of gastroenteropancreatic neuroendocrine tumours with octreotide LAR. AB - BACKGROUND: Octreotide long acting repeteable (LAR) is a new somatostatin analogue whose activity lasts 28 days. AIM: To assess its therapeutic efficacy, tolerability, and safety in patients with gastroenteropancreatic neuroendocrine tumours. METHODS: A total of 16 patients were studied; 10 patients with carcinoid tumours, three with non-functioning pancreatic tumours, two with Zollinger Ellison syndrome associated with multiple endocrine neoplasia type 1, and one with glucagonoma were studied. Octreotide LAR was administered intramuscularly at a dose of 20 mg every 28 days for a mean of 10.7 months (range 6-15 months). RESULTS: In carcinoid tumour patients, octreotide LAR normalized bowel movements in nine out of 10 cases, and flushing episodes disappeared in seven out of eight cases. Even in the remaining six patients the symptoms disappeared. In carcinoid tumour patients, urinary 5-hydroxyindoleacetic acid decreased significantly. In the two patients with Zollinger-Ellison syndrome/multiple endocrine neoplasma type 1 and in the patient with glucagonoma, serum gastrin and plasma glucagon, respectively, decreased considerably. Tumour size remained unchanged in 14 out of 16 patients, and increased in the remaining two. No side-effects were observed. CONCLUSIONS: Octreotide LAR appears to have a good therapeutic efficacy, tolerability and safety in the treatment of neuroendocrine tumours. Its effects are similar to those of octreotide and lanreotide. However, because it only needs to be administered once every 28 days, it is preferable in clinical practice. PMID- 10792121 TI - Beta(beta)3-adrenergic receptors in human pancreatic islet and duodenal somatostatin neuroendocrine cells. AB - BACKGROUND: We previously localized beta3-adrenergic receptors immunohistochemically in human gastrointestinal smooth muscle and incidently found a population of human pancreatic islet cells and duodenal epithelial neuroendocrine cells that also expressed beta3-adrenergic receptors. AIM: To identify the nature of the islet and duodenal cells that stained positive for beta3-adrenergic receptors. METHODS: Paraffin sections of human pancreas and duodenum and Chinese hamster ovary cells transfected with the human beta3 adrenergic receptor were immuno-stained for beta3-adrenergic receptors using an affinity-purified rabbit polyclonal antibody (anti-P12) raised against a 15 amino acid sequence (P12) of the human receptor. Immunohistochemical staining for the receptor was carried out in the presence and absence of P12 peptide and both somatostatin 14 and 18 peptides. beta3-adrenergic receptor-stained sections were also double-immunostained with anti-insulin, -glucagon, -somatostatin and pancreatic polypeptide antibodies. RESULTS: A subpopulation of human pancreatic islet cells and duodenal epithelial cells expressed positive cytoplasmic beta3 adrenergic receptor immunostaining. Using distribution and double-staining techniques, these cells were found to be somatostatin-positive D cells but not A or B cells. The positive staining of D cells with anti-P12 antibody was inhibited by prior incubation of the antibody with P12 peptide but not somatostatin-14 or 28 peptides. Pancreatic vascular smooth muscle and duodenal vascular and non vascular smooth muscle also stained with anti-P12 antibody. Transfected Chinese hamster ovary cells showed positive membrane staining. CONCLUSION: We have identified a population of neuroendocrine cells in the human pancreas and duodenum that express beta3-adrenergic receptors. These cells appear to be somatostatin D cells. PMID- 10792120 TI - Effects of midazolam on small bowel motility in humans. AB - BACKGROUND: Benzodiazepines are used as sedatives for some intestinal procedures and as hypnotics, and this is the reason for studying their effects on duodenojejunal motility. METHODS: Antroduodenojejunal manometry was performed in 13 healthy volunteers on two different occasions, when placebo or midazolam were given intravenously (randomized, double-blind). A bolus dose of midazolam 0.03 mg/kg was followed by 0.015 mg/kg after 1.5, 3 and 4.5 h. After 5 h observation of interdigestive motility, the volunteers were given a test meal and recording continued for another hour. Twenty-eight motility variables were compared. RESULTS: With midazolam the median motility index of phase III in the proximal duodenum was increased by 37% (P < 0.05), which was a consequence of both a longer duration (P < 0.01) and higher pressure amplitudes (P < 0.05), compared with placebo. A longer duration (9%) of phase III was also seen in the distal duodenum (P < 0.05). With midazolam the duration of the migrating motor complex was shortened by 27% (P < 0. 05). No statistically significant difference was found for the number of episodes of phase III registered (P=0.09), or for the other 22 motility variables compared including the duodenal retroperistalsis in late phase III. CONCLUSION: Midazolam does affect some aspects of duodenal motility, especially in the proximal part, but phase III-related retroperistalsis is not affected. PMID- 10792122 TI - The osmotic laxative magnesium sulphate activates the ileal brake. AB - BACKGROUND: Alterations in gastrointestinal motility and hormone secretion, especially activation of the ileal brake, have been documented in malabsorption. AIM: To investigate whether artificially-induced accelerated small intestinal transit activates the ileal brake mechanism. METHODS: Eight healthy volunteers (four female, four male; age 21 +/- 3 years) participated in four experiments: (a) meal with either oral magnesium sulphate (MgSO4) or placebo; and (b) fasting with either oral MgSO4 or placebo. Antroduodenal motility was recorded by perfusion manometry. Duodenocaecal transit time was determined by the lactulose H2 breath test. Gall-bladder volume was measured by ultrasound at regular intervals, and blood samples were drawn for determination of cholecystokinin and peptide YY (RIA). Twenty-four hour faecal weight and fat excretion were determined. RESULTS: MgS04 significantly accelerated duodenocaecal transit time and increased faecal fat and weight in all subjects. MgSO4 significantly delayed the reoccurrence of phase III and affected antroduodenal motility during fasting but not after meal ingestion. Postprandial gall-bladder relaxation and postprandial peptide YY release were significantly increased during the MgSO4 experiment compared to placebo. CONCLUSIONS: The osmotic laxative MgS04 accelerates intestinal transit both in the fasting and fed state. MgS04 activates the ileal brake mechanism only in the fed state, with peptide YY release and inhibition of gall-bladder emptying. PMID- 10792123 TI - Failure of oesophageal acid control in candidates for Barrett's oesophagus reversal on a very high dose of proton pump inhibitor. AB - BACKGROUND: Normalization of oesophageal acid exposure using high dose proton pump inhibitors in patients who are candidates for ablation therapy has been suggested to be essential for successful Barrett's reversal. However, the success rate for achieving pH normalization has not been determined. METHODS: Patients with Barrett's oesophagus (2-6 cm in length) who were found to be eligible for ablation therapy using multipolar electrocoagulation were included in this prospective study. Patients underwent an upper endoscopy to determine Barrett's length and other anatomic characteristics. Biopsies were obtained to rule out dysplasia. Subsequently, patients were treated with omeprazole 40 mg b.d. Twenty four hour oesophageal pH monitoring was performed after a mean period of 8.4 +/- 0.6 days of therapy. RESULTS: Twenty-five patients were enrolled into the study. The pH test was abnormal in four (16%) of the study subjects. An additional two (8%) patients had abnormal supine percentage time with pH less than 4. There was no significant difference in oesophageal acid control between patients with long vs. short segment Barrett's oesophagus. Elderly (> 60 years) patients tended to have less acid control than younger (/= 97% in all patients. PMID- 10792138 TI - The bacterial and viral filtration performance of breathing system filters. AB - The bacterial and viral filtration performance of 12 breathing system filters was determined using test methods specified in the draft European standard for breathing system filters, BS EN 13328-1. All the filters were of two types, either pleated hydrophobic or electrostatic, and these two types differed in their filtration performance. The filtration performance is expressed in terms of the microbial penetration value, defined as the number of microbes passing through the filter per 10 million microbes in the challenge. The geometric mean (95% confidence limits) microbial penetration value was 1.0 (0.5, 3.5) and 2390 (617, 10 000) for the pleated hydrophobic and electrostatic filters, respectively, for the bacterial challenge, and 87 (48, 212) and 32 600 (10 900, 84 900), respectively, for the viral challenge. In general, there was little change in the microbial penetration values following 24 h simulated use. It is concluded that results from the tests specified in the draft standard will allow comparisons to be made between different manufacturers' products enabling an informed choice to be made. PMID- 10792139 TI - Microbial contamination of gum elastic bougies. AB - The gum elastic bougie is a simple device that is used to assist in the management of the difficult intubation. It is not uncommon for a bougie to be re used many times. This study investigated the incidence of microbial contamination of the bougies in one hospital. Potentially pathogenic organisms were identified both on the bougies and in their storage containers. This has implications for their cleaning and maintenance, and raises the question as to whether we should replace them with single-use, disposable devices. PMID- 10792140 TI - Pharyngeal abscess presenting with upper airway obstruction and atlanto-axial subluxation in a small infant. AB - We describe an infant who presented with a combination of upper airway obstruction and atlanto-axial subluxation, secondary to a pharyngeal abscess resulting from cosmetic ear piercing. This combination posed a number of difficulties for the anasthetist and a detailed plan was formulated to prepare the child for anasthesia. PMID- 10792141 TI - Anaesthesia in myotubular (centronuclear) myopathy. AB - A patient with a known history of myotubular myopathy presented for surgery for insertion of a tibial nail. Anasthesia was induced and maintained using an intravenous anasthetic technique. Neuromuscular function was assessed using mechanomyography, which showed a profound reduction in muscle contractility. In view of this, the use of muscle relaxants was avoided altogether. Nerve conduction was normal but electromyography showed small motor units, with generalised distribution, suggesting mild to moderately severe myopathy. The patient made a slow but uneventful recovery. PMID- 10792142 TI - Fentanyl supplementation of sevoflurane induction of anaesthesia. AB - Sevoflurane induction of anasthesia has been examined extensively, but little is known about the usefulness of other drugs as adjuncts to hasten and smooth the process. Sixty patients, undergoing surgery of a type suitable for a spontaneous respiration, laryngeal mask airway anasthetic technique, were randomly allocated to receive 1.0 microgram.kg-1 intravenous fentanyl or the equivalent volume of normal saline, 30 s prior to triple-breath induction with sevoflurane. The study was double-blind. There were no differences between the groups for the times to loss of eyelash reflex, jaw relaxation, insertion of the laryngeal mask airway or regular settled breathing. However, there was a difference in the incidence of adverse airway events (breath-holding, coughing and laryngospasm) between the two groups (16.5% in the fentanyl group and 40% in the placebo group); this did not reach statistical significance. Both groups were haemodynamically stable throughout induction, although the fentanyl group had a statistically significant decrease in systolic blood pressure at 4 min compared with the placebo group, which was not considered clinically relevant. We conclude that fentanyl has no significant influence over the speed and quality of sevoflurane induction. PMID- 10792143 TI - An investigation into the quantity of secretions removed by inflated and deflated laryngeal mask airways. AB - It has been suggested that removal of a laryngeal mask airway with the cuff inflated may remove more secretions than with the cuff deflated. We performed a study to determine whether this suggestion is correct. Patients were randomly allocated to removal of the laryngeal mask airway with the cuff deflated (n = 75) or inflated (n = 74). The laryngeal mask airways were weighed before insertion and after removal, the difference in these two weights being taken to be the mass of secretions adherent to the airways on removal. The mean (SD) increase in laryngeal mask airway weight was 2.45 (1.47) g with the cuff deflated and 3.03 (1.76) g with the cuff inflated (p = 0.03). We conclude that removal of the laryngeal mask airway with the cuff inflated removes approximately 0.5 g more secretions than with the cuff deflated. PMID- 10792144 TI - Carotid endarterectomy. A survey of UK anaesthetic practice. AB - We conducted a postal questionnaire survey of the members of the Vascular Anasthesia Society of Great Britain and Ireland, asking questions about the provision of anasthesia for carotid endartectomy. Of 215 respondents, 187 were currently providing anasthesia for carotid endarterectomy. The majority of respondents (69%) always use general anasthesia for this operation but 99/215 (46%) had some experience of regional anasthesia for carotid endartectomy. Amongst those currently using regional anasthesia, combined deep and superficial cervical plexus block was the technique used by 71%. Other regional techniques used included local infiltration and superficial block alone. During regional anasthesia, most (66%) anasthetists used cerebral monitoring techniques such as stump pressure or transcranial Doppler as well as keeping the patient awake. However, in a significant proportion of cases (37%) under general anasthesia no cerebral monitoring was used. Reported surgical shunt insertion rates were lower in awake (mean 42%) patients than those receiving general anasthesia (61%). Respondents using regional anasthesia were more likely to feel that their technique was appropriate than those using general anasthesia. PMID- 10792145 TI - Difficult airway equipment in English emergency departments. AB - The need for tracheal intubation in the emergency department is often unpredictable and precipitous in nature. When compared with the operating room, a higher incidence of difficult intubation is observed. There are currently no accepted guidelines with respect to the stocking of difficult airway equipment in the emergency department. We have conducted a telephone survey to determine the availability of equipment for the management of the difficult airway in English emergency departments. Overall, the majority of units held a curved laryngoscope blade (100%), gum elastic bougie (99%) and surgical airway device (98%). Of alternative devices for ventilation, a laryngeal mask airway was kept by 65% of departments, a needle cricothyroidostomy kit by 63% and an oesophageal-tracheal twin-lumen airway (Combitube) by 18%. Of alternative devices for intubation, fewer than 10% held a retrograde intubating kit, intubating laryngeal mask, bronchoscope or lighted stylet. Seventy-four per cent of departments held an end tidal carbon dioxide detector. PMID- 10792146 TI - Transfer of critically ill patients in the Northern region. PMID- 10792147 TI - Persistent pruritus after pentastarch infusions in intensive care patients. PMID- 10792148 TI - Cardiac arrest after video-assisted thoracoscopic drainage of pleural effusion. PMID- 10792149 TI - Unilateral pulmonary oedema following general anaesthesia--previous chest wall irradiation as a possible contributing factor. PMID- 10792150 TI - Continuous or intermittent loss of resistance for identifying the epidural space. PMID- 10792151 TI - Epidural analgesia for Caesarean section. PMID- 10792152 TI - The effectiveness of tenoxicam vs isosorbide dinitrate plus tenoxicam in the treatment of acute renal colic. AB - OBJECTIVE: To determine the effectiveness of isosorbide dinitrate in combination with tenoxicam compared with tenoxicam alone for the relief of acute renal colic. PATIENTS AND METHODS: Fifty patients with acute renal colic were randomly and in a double blind fashion divided into two treatment groups of 25 each. Group 1 received tenoxicam (40 mg intravenously) and group 2 received tenoxicam (40 mg intravenously) plus sublingual isosorbide dinitrate (5 mg). The patients were compared for visual pain scores, heart rate and blood pressure between and within the groups before and after treatment. RESULTS: Comparing the groups, group 2 had significantly lower pain scores after treatment (P < 0.05) but no other variables were significantly different (P > 0.05 for each). Within the groups the pain scores were significantly lower after treatment in both groups (P < 0.05). Group 2 also had a significantly lower heart rate and blood pressure after treatment (P < 0.05). There were no side-effects caused by tenoxicam and isosorbide dinitrate in either group. CONCLUSION: The use of tenoxicam alone or combined with isosorbide dinitrate was effective in relieving of renal colic, but the relief obtained with the combination was significantly greater than tenoxicam alone. PMID- 10792153 TI - The relative incidence of detrusor instability and bacterial cystitis detected on the urodynamic-test day. AB - OBJECTIVE: To determine whether patients with detrusor instability (DI) were more likely to have bacterial cystitis or significant bacteriuria on the urodynamic test day than were women with a stable bladder. PATIENTS AND METHODS: A catheter specimen of urine was cultured (overnight in air) from 862 consecutive women at the time of urodynamic testing. The upper urinary tract was imaged, with cystoscopy when indicated, to exclude upper tract lesions or malignancy. The percentage of patients with pure idiopathic DI and those with mixed DI/genuine stress incontinence (GSI), in whom the urine culture was positive, was compared with the percentage who had a stable bladder (pure GSI or urodynamically normal) and a positive urine culture, both for the entire dataset and for women aged > or <65 years. Data were also analysed to detect the converse relationship; in those women found to have bacterial cystitis, the relative risk of being found urodynamically unstable or stable was determined. RESULTS: The likelihood of bacterial cystitis occurring in patients with idiopathic DI (5.6%) was significantly greater than that in patients with GSI (1.1%; P = 0.009, Fisher's exact test). The proportion of patients with DI and significant bacteriuria (15. 4%) was significantly greater than that in patients with GSI (7.9%; P = 0.02). In patients with combined pure and mixed DI, bacterial cystitis was significantly more likely to occur (6.3%) than in GSI (P < 0.001), but bacteriuria was no more likely (12.5%, P = 0.09). Conversely, of those women found to have bacterial cystitis, the relative risk of having an unstable bladder was increased (+1.56), but for those with bacteriuria the relative risk of detrusor instability was not increased. CONCLUSION: There was a significant association between idiopathic DI and bacterial cystitis, and we suggest that in some women with an unstable bladder, urinary infection may enhance detrusor contractility. Nevertheless, large-scale studies are needed of the temporal relationship between the onset of bacterial cystitis and the onset of DI. PMID- 10792154 TI - A randomized controlled trial comparing the efficacy of controlled-release oxybutynin tablets (10 mg once daily) with conventional oxybutynin tablets (5 mg twice daily) in patients whose symptoms were stabilized on 5 mg twice daily of oxybutynin. AB - OBJECTIVE: To compare the efficacy of a controlled-release (CR) formulation of oxybutynin with that of conventional oxybutynin in patients with detrusor instability or detrusor hyper-reflexia whose symptoms were stabilized on conventional oral oxybutynin tablets. PATIENTS AND METHODS: The study comprised a randomized, double-blind, parallel-group trial involving 130 patients drawn from 15 centres in the UK. The study was of 6 weeks' duration, i.e. 2 weeks of screening whilst taking conventional oxybutynin tablets (5 mg twice daily) followed by 4 weeks of double-blind treatment with either CR oxybutynin tablets (10 mg once daily) or conventional oxybutynin tablets (5 mg twice daily). Outcome measures were changes in 24-h frequency and 24-h incontinence episodes recorded throughout the study on diary charts. Adverse events were recorded by patients in their diary charts and serum concentrations of oxybutynin and its active metabolite, N-desethyloxybutynin, were measured at baseline and at completion of the study to detect possible drug accumulation. RESULTS: The treatments did not differ significantly in any of the outcome measures. The primary efficacy criterion was the daytime continence at completion of the study; 53% and 58% of patients were continent on CR and conventional oxybutynin treatments, respectively (the 95% confidence interval of the difference in the proportion being - 22% to 13%; P = 0.62). The total number of side-effects experienced by those patients receiving treatment with the CR formulation was 57% of that for patients receiving treatment with conventional oxybutynin. Individual side effects showed a similar distribution within treatment groups. There was no evidence of the accumulation of oxybutynin or N-desethyloxybutynin during the multiple dosing of CR or conventional oxybutynin tablets. CONCLUSION: The CR and conventional formulations of oxybutynin did not differ in their efficacy, and the CR formulation was associated with fewer side-effects. In addition, CR oxybutynin appeared to maintain therapeutic blood levels over the 24 h dosing interval with no accumulation of oxybutynin or its active metabolite. Once-daily dosing with a CR tablet is seen as convenient for the patient and is expected to result in improved compliance in patients already stabilized on conventional oxybutynin treatment. PMID- 10792155 TI - Bacillus Calmette-Guerin instillation treatment for carcinoma in situ of the upper urinary tract. AB - OBJECTIVE: To analyse the efficacy and safety of bacillus Calmette-Guerin (BCG) perfusion treatment forcarcinoma in situ (CIS) of the upper urinary tract. PATIENTS AND METHODS: Six patients with cytologically diagnosed CIS of the upper urinary tract were treated by BCG instillation via retrograde catheterization using a 6 F ureteric catheter or an 8 F indwelling JJ ureteric stent between the ureter and bladder. BCG (Tokyo 172 strain, 80 mg in 100 mL normal saline) was instilled weekly for 4 or 8 weeks. The efficacy and safety of the treatment was assessed. RESULTS: The mean (range) follow-up was 22 (9-38) months; none of the patients died, and all were negative for cytology in urine collected from the upper urinary tracts. However, one patient had recurrent CIS in the prostatic urethra; the patient was treated by intravesical BCG instillation. In all patients, positive cytology became negative after one or two instillations of BCG. The ureter became stenotic in two patients. Although irritative symptoms occurred in all patients, such side-effects disappeared in a few months and were not clinically significant. CONCLUSION: In these six patients retrograde BCG instillation for CIS of the upper urinary tract was effective and safe. Based on the cytology results after only two BCG treatments, the dose or number of BCG instillations could possibly be reduced, but further study is needed. PMID- 10792156 TI - Urothelium-preserving augmentation cystoplasty covered with a peritoneal flap. AB - OBJECTIVES: To present the results of bladder autoaugmentation covered with a peritoneal flap in patients with bladder dysfunction. PATIENTS AND METHODS: Thirteen patients (seven male and six female, mean age 11.9 years, range 4-25) who underwent autoaugmentation covered with a peritoneal flap were evaluated. Seven had different forms of myelodysplasia, four had spinal cord injury and two had Hinman syndrome as the cause of bladder dysfunction. Indications for augmentation included upper tract deterioration, urinary incontinence and recurrent urinary tract infection, despite anticholin-ergic therapy. RESULTS: The mean bladder capacity increased by 18.6% after surgery and the mean compliance at capacity increased from 3.4 to 5.8 cmH2O/mL. All patients were incontinent before surgery and continence was achieved in only six afterward. Four patients showed no clinical or urodynamic improvement and required re-augmentation using intestinal segments. Only three patients needed no anticholinergic therapy after surgery. All four patients in whom the procedure failed had capacities of < 30% of that expected for their age. There were no metabolic problems. CONCLUSION: Autoaugmentation combined with a peritoneal flap is an easy procedure but the clinical results are poor in some patients, especially those with a small initial bladder capacity. The need for secondary augmentation with enteric segments was common. The use of a peritoneal flap does not appear to increase the capacity and compliance more than is obtained with the classical technique; it may prevent adhesion to the abdominal wall and make a secondary procedure easier. As the increase in capacity and compliance is limited with this technique, a urothelium preserving augmentation should be reserved for those bladders with a relatively good initial capacity. PMID- 10792157 TI - Are dedicated bladder films necessary as part of intravenous urography for haematuria? AB - OBJECTIVE: To determine the use and assess the value of full-bladder films during intravenous urography (IVU) which, despite the widespread availability of flexible cystoscopy, remain part of IVU in many radiology departments. Materials and methods A telephone survey of all Scottish radiology departments where IVU is regularly used showed that half routinely included a full-bladder film in the series. The reports of all IVU over 2 years in the authors' department were analysed retrospectively. The index urogram of all patients with bladder tumours confirmed during this period was reviewed independently by three observers, and together with the initial radiological reports was correlated with the cystoscopic and histological findings. RESULTS: From 2625 patients, 139 (5.2%) IVU reports commented on the bladder; 1423 patients presented with no haematuria. None of the patients without haematuria, where a comment was made about the bladder, had pathological evidence of a tumour. Overall 121 of 464 (26%) new bladder tumours were diagnosed on IVU before cystoscopy. Multiple tumours were always undetected and large tumours were often overlooked. CONCLUSIONS: Despite its continuing popularity, IVU is a poor means of identifying bladder tumours and routine views of the full bladder should be abandoned. PMID- 10792158 TI - A predictive model of survival after radical cystectomy for carcinoma of the bladder. AB - OBJECTIVE: To evaluate the effect of patient and tumour characteristics on the disease-free survival after radical cystectomy for infiltrating bladder cancer, and to use these to help in constructing a meaningful prognostic index. METHODS: The disease-free survival was initially evaluated in 1026 patients (the reference series, 1969-1990). A multivariate analysis showed that the tumour P stage, grade and nodal involvement were the only factors which had an independent and significant association with survival. The computed regression coefficients were then used to classify patients into one of four risk categories and the results then validated by applying the model to a prospective test series (1991-1995). RESULTS: The 5-year disease-free survival of both groups was similar. When the results for the risk categories of the reference series were compared with those of the test series, there was no significant difference. CONCLUSION: This comprehensive prognostic model for the results of radical cystectomy was validated and verified in a prospective group of patients. Adjuvant therapies are indicated for patients with a high risk score. PMID- 10792159 TI - Lymphadenectomy in patients with transitional cell carcinoma of the urinary bladder; significance for staging and prognosis. AB - OBJECTIVE: To determine the need to standardize the number and location of lymph nodes to be removed during radical cystectomy in patients with invasive bladder carcinoma. PATIENTS AND METHODS: The pelvic lymph nodes from 447 patients (mean age 62.8 years) who underwent radical cystectomy between 1986 and 1997 were evaluated. The number of lymph nodes was correlated with the depth of invasion of the primary tumour (pT), occurrence of nodal metastases, clinical outcome, the operating surgeons and the pathologists dissecting the nodes. RESULTS: The clinical follow-up was available for 302 patients (mean follow-up 38.7 months). The mean (range) number of lymph nodes removed was 14.7 (1-46). The number of lymph nodes removed varied significantly among different surgeons but not among pathologists. In pT3 and pT4 tumours, a more extended lymphadenectomy (>/= 16 lymph nodes) correlated with a higher percentage of patients with documented nodal metastases. There was a significant correlation between the number of lymph nodes removed and the tumour-free 5-year survival in patients with pT1, pT2 or pT3 tumours, and in patients with 1-5 positive lymph nodes (P < 0.01). CONCLUSION: Extensive lymphadenectomy significantly improves the prognosis of patients with invasive bladder cancer and represents a potentially curative procedure in patients with nodal metastases, including micrometastases that may escape detection during routine histopathological evaluation. The results indicate the need for a standardized lymph node dissection. PMID- 10792160 TI - Long-term follow-up of noninvasive bladder tumours (stage Ta): recurrence and progression. AB - OBJECTIVE: To evaluate long-term recurrence-free and progression-free survival of noninvasive bladder tumours (stage Ta), and the significance of simple risk factors, including concomitant epithelial dysplasia. PATIENTS AND METHODS: The study included 217 patients with primary noninvasive bladder tumour (stage Ta) who were followed routinely for up to 20 years. Voided urine cytology (VUC) and preselected site biopsies (PSB) were obtained prospectively to evaluate the significance of concomitant epithelial dysplasia. RESULTS: The mean follow-up was 84 months (maximum 238). Of all tumours, 39% did not relapse, a further 20% recurred infrequently (less than once a year) and 41% recurred frequently, amongst which the most frequent were multiple and early recurrent tumours; 42 (19%) tumours progressed to stage T1+ and 23 (11%) progressed further (stage T2+ or metastases). No grade 1 tumours became invasive. Positive VUC or PSB, a short recurrence-free period or multiplicity, and size > 3 cm were significant predictive factors. The treatment and surveillance of epithelium-confined bladder tumours are discussed. CONCLUSION: Concomitant dysplasia and early recurrence are associated with considerable risk of progression in the long-term follow-up in a group of otherwise low-risk superficial bladder tumours (stage Ta). PMID- 10792161 TI - Bcl-2 expression identifies patients with advanced bladder cancer treated by radiotherapy who benefit from neoadjuvant chemotherapy. AB - OBJECTIVE: To assess the prognostic significance of Bcl-2 expression on the clinical outcome after radiotherapy for muscle-invasive bladder cancer, and to determine if it is possible to identify a subgroup of patients to whom neoadjuvant chemotherapy can be targeted to improve survival. PATIENTS AND METHODS: Immunohistochemical staining for Bcl-2 and p53 was performed on the tumours of 51 patients with stage T2-T4a NXM0 transitional cell carcinoma of the bladder who had been included in a randomized clinical trial of radiotherapy with or without neoadjuvant cisplatin. The association between positive staining and salvage cystectomy rate and overall survival was examined, with a median follow up of 12 years. RESULTS: Bcl-2 and p53 expression was positive in 31 (61%) and 39 (76%) of the tumours, with no association between either, or with tumour stage or grade. There was no difference according to Bcl-2 positivity in the salvage cystectomy rate (P = 0.83) or survival (P = 0.68) for the 51 patients as a whole, but Bcl-2-negative patients receiving neoadjuvant cisplatin had a significantly better prognosis, with a median survival of 72 months compared to 17 months in Bcl-2-positive patients, and a 5-year survival rate of 55% (P = 0.03). CONCLUSIONS: Quantifying Bcl-2 in patients undergoing radiotherapy for advanced bladder cancer identifies those who may benefit from neoadjuvant chemotherapy. Further studies of other members of the Bcl-2 family and other proteins controlling both cell proliferation and apoptosis are warranted, to define the roles and the interactions between them that may contribute to oncogenesis and resistance to standard treatments. This may allow the targeting of specific treatments to patients known to be sensitive to them, and aid the future development of novel therapies for bladder cancer. PMID- 10792162 TI - A systematic review of Cernilton for the treatment of benign prostatic hyperplasia. AB - OBJECTIVE: To systematically review the evidence for the clinical effects and safety of the rye-grass pollen extract (Cernilton) in men with symptomatic benign prostatic hyperplasia (BPH). METHODS: Trials were identified by searching Medline, specialized databases (EMBASE, Cochrane Library, Phytodok), bibliographies, and contacting relevant trialists and manufacturers. Randomized or controlled clinical trials were included if: men with symptomatic BPH were treated with Cernilton; a control group received either placebo or pharmacological therapy; the treatment duration was >/= 30 days; and clinical outcomes were reported. RESULTS: In all, 444 men were enrolled in two placebo controlled and two comparative trials lasting 12-24 weeks. Three studies used a double-blind method although the concealment of treatment allocation was unclear in all. Cernilton improved 'self-rated urinary symptoms' (the proportion reporting satisfactory or improving symptoms) vs placebo and another plant product, Tadenan. The weighted mean (95% confidence interval) risk ratio (RR) for self-rated improvement vs placebo was 2.40 (1. 21-4.75) and the weighted RR vs Tadenan was 1.42 (1.21-4.75). Cernilton reduced nocturia compared with placebo or Paraprost (a mixture of amino acids); against placebo, the weighted RR was 2.05 (1.41-3.00), and against Paraprost the weighted mean difference for nocturia was 0.40 times per evening (- 0.73 to 0.07). Cernilton did not improve urinary flow rates, residual volume or prostate size compared with placebo or the comparative study agents. Adverse events were rare and mild; the withdrawal rate for Cernilton was 4. 8%, compared with 2.7% for placebo and 5.2% for Paraprost. CONCLUSIONS: The Cernilton trials analysed were limited by their short duration, limited number of enrolees, omissions in reported outcomes, and the unknown quality of the preparations used. The comparative trials had no confirmed active control. The available evidence suggests that Cernilton is well tolerated and modestly improves overall urological symptoms, including nocturia. Additional randomized placebo and active-controlled trials are needed to evaluate the long term clinical effectiveness and safety of Cernilton. PMID- 10792163 TI - Treatment of symptomatic benign prostatic hyperplasia with beta-sitosterol: an 18 month follow-up. AB - OBJECTIVES: To determine the long-term effects of phytotherapy with beta sitosterol (the trade name for beta-sitosterol used in this study is Harzol(R)) for symptomatic benign prostatic hyperplasia (BPH). Patient and methods At 18 months after enrolment in a 6-month multicentre double-blind placebo-controlled clinical trial with beta-sitosterol (reported previously), patients were re evaluated using the modified Boyarsky score, the International Prostate Symptom Score and quality-of-life index, the maximum urinary flow rate (Qmax) and postvoid residual urine volume (PVR). In this open extension of the original trial (after 6 months of treatment or placebo), patients were free to chose their further treatment for BPH. RESULTS: In all, 117 patients (59%) were eligible for analysis during the follow-up. Of the formerbeta-sitosterol group, 38 patients who continued beta-sitosterol treatment had stable values for all outcome variables between the end of the double-blind study and after 18 months of follow up. The 41 patients choosing no further therapy had slightly worse symptom scores and PVR, but no changes in Qmax. Of the former placebo group, 27 patients who started beta-sitosterol after the double-blind trial improved to the same extent as the treated group for all outcome variables. The 18 patients choosing no further therapy showed no signs of improvement. CONCLUSION: The beneficial effects of beta-sitosterol treatment recorded in the 6-month double-blind trial were maintained for 18 months. Further clinical trials should be conducted to confirm these results before concluding that phytotherapy with beta-sitosterol is effective. PMID- 10792164 TI - The short-term prostate-specific antigen velocity before biopsy can be used to predict prostatic histology. AB - OBJECTIVES: To evaluate whether the short-term prostate-specific antigen (PSA) velocity before biopsy can be used to predict prostatic histology, and to assess the role of a second PSA measurement before prostate biopsy. PATIENTS AND METHODS: The study comprised 197 patients who were referred for transrectal ultrasonography (TRUS) and prostate biopsy. The PSA level was initially measured at the first outpatient assessment; patients with a serum PSA level of < 4 ng/mL and > 50 ng/mL were excluded. Blood samples were taken just before prostate biopsy for the second PSA measurement. The mean interval between the measurements was 2.2 months. The short-term PSA velocity was calculated and correlations between this variable and age, prostate volume and initial PSA levels determined. RESULTS: There was a statistically significant difference between the short-term PSA velocity of patients with benign prostate histology and those with prostate cancer (P < 0.05). The short-term PSA velocity alone had the same diagnostic accuracy as the serum PSA level (area under the receiver-operating characteristic curve 0.612). There was only a weak correlation between the short-term PSA velocity and prostate volume. However, there was no correlation with age and initial PSA level in patients with benign histology. The second PSA measurement had higher specificity without losing sensitivity. CONCLUSION: The short-term PSA velocity estimated before biopsy can be used to predict prostatic histology. By measuring serum PSA 2 months after the first in patients with serum PSA level of 4-10 ng/mL, the number of negative biopsies can be reduced by 17%. PMID- 10792165 TI - Surgical conversion of genitalia in transsexual patients. AB - OBJECTIVE: To describe the techniques and outcome of genital and urethral reconstructive surgery during gender conversion as part of the treatment of transsexuals. PATIENTS AND METHODS: From 1992 to 1999, 82 patients were surgically converted after previous sexual and hormonal therapy. Using the male genital tissue to create new female genitalia, and vice versa, 30 male and 52 female transsexuals were converted. For male-to-female transsexuals, the technique of penile skin inversion was used 29 times and sigmoidocolpoplasty five times (in one patient primarily and in four patients to correct inadequate neovaginal size after penile skin inversion). In female-to-male transsexuals, 28 meta-idoioplasties and seven neophalloplasties were performed using the groin skin-flap technique, with 42 breast reductions also included as a part of the therapy. RESULTS: Surgical gender reassignment of the male transsexuals resulted in replicas of female genitalia which enabled coitus with orgasm. Depending on the technique used in the reverse conversion, the patient maintained the ability to attain orgasm, and in many cases had a satisfactory appearance of the neopenis, with the potential to void while standing. CONCLUSIONS: The morphological proportions of each patient vary, and the different shapes and sizes of the tissues can be used for plastic operations. Thus the modelling of each individual genital in transsexuals can be considered 'original'. PMID- 10792166 TI - Reflex innervation of the bulbocavernosus muscle. AB - OBJECTIVE: To determine the somatic reflex innervation of the bulbocavernosus muscle (BCM) and to illustrate the role of the perineal nerve in BCM contraction, using electrophysiological techniques. Subjects and methods Ten healthy, potent men underwent genital electrophysiological testing. Stimuli were applied to the dorsal nerve of the penis at the base of the penis, glans and anterior urethra, and the perineal nerve branch to the BCM. The electromyographic (EMG) responses of the BCM were recorded and onset latencies measured at baseline, both after anaesthetising the perineal nerve and after recovery from the anaesthetic. RESULTS: A reflex BCM contraction was elicited from all stimulation sites. Anaesthetizing the perineal nerve resulted in alterations of the EMG response. After the anaesthetic effect wore off, the responses returned to baseline values. CONCLUSIONS: Three distinct somatic bulbocavernosus reflexes were detected which are components of normal ejaculation. BCM contraction can be elicited after stimulating the dorsal penile nerve at the glans and the anterior urethra; these are the flexor responses of the bulbocavernosus reflex. BCM contraction can also be induced on stimulating the perineal nerve, which is the pathway through which the stretch and the tendon organ reflexes are carried to mediate muscle tone. The motor innervation of the BCM for all reflexes is carried through a branch of the perineal nerve. The findings may be relevant to the evaluation of ejaculatory disorders. PMID- 10792167 TI - Bilateral germ cell cancer of the testis: a report of 11 patients with a long term follow-up. AB - OBJECTIVE: To describe the incidence, clinical characteristics, treatment methods and long-term follow-up of bilateral germ cell tumours of the testis (GCTT) in patients treated at one institution. PATIENTS AND METHODS: Of 552 patients with GCTT, 11 (2%, mean age 26. 9 years) developed bilateral disease; all 11 underwent radical orchidectomy. Additional treatment was planned according to the histological type and clinical stage of the tumour, and previous treatments. Intramuscular testosterone was administered periodically after total castration. The data on survival, sexual status and treatment complications were reviewed. RESULTS: Of the 11 patients, seven developed a second tumour metachronously (median interval 87 months) and four had synchronous bilateral GCTT. Cryptorchidism, infertility or atrophic testis was associated with the development of bilateral GCTT in seven of the 11 patients. All synchronous tumours and most of the sequential tumours had identical histology on both sides. Although all sequential tumours presented at an early clinical stage, three of four synchronous bilateral GCTTs presented at an advanced stage. Five patients received platinum-based chemotherapy; three patients underwent post- chemotherapy resection of the retroperitoneal residual mass. Sexual libido and potency were conserved in all patients. No significant morbidity was recorded as being caused by any of these treatments. At a median follow-up of 11. 6 years, all patients were alive with no evidence of cancer. CONCLUSIONS: All patients with unilateral GCTT have an increased risk of developing a contralateral testicular tumour, even decades after diagnosis. Management should be adapted to each patient. As all patients in this series survived in the long-term, developing a second germ cell cancer does not necessarily predict a poor prognosis. PMID- 10792168 TI - An epidemiological study of enuresis in Korean children. AB - OBJECTIVE: To estimate the prevalence of enuresis in children of elementary school age, to evaluate the impact of enuresis on these children and their parents, and to identify the methods and effectiveness of managing enuresis. Subjects and methods A randomly selected cross-sectional study was conducted in one elementary school in each urban ward (nine schools) in Pusan, Korea. The parents of these 12 570 children aged 7-12 years were asked to complete questionnaires which included items about the presence and frequency of enuresis, its perceived impact and management. Enuresis was defined as an episode of wetting occurring at least once per month. RESULTS: The overall response rate to the questionnaire was 55.8% (girls 28.2%, boys 27.6%). The prevalence of nocturnal, diurnal and combined enuresis was 9.2%, 2.2% and 1.4%, respectively. The overall prevalence of enuresis declined with age from 20.4% at 7 years old to 5.6% at 12 years old; 342 (57.0%) parents and 318 (55. 6%) children were concerned about enuresis. The common self-help strategies were waking the child at night to void (38.1%) and restriction of water intake (25.7%). Of the enuretic children, only 13.7% had consulted a health worker. CONCLUSION: The prevalence rates for enuresis in Pusan are similar to those reported from European countries. Enuretic children and their parents were moderately concerned about enuresis and the parents primarily used self-management within the family. PMID- 10792169 TI - Paediatric anatrophic nephrolithotomy; stone clearance - at what price? AB - OBJECTIVE: To evaluate the functional outcome of anatrophic nephrolithotomy in children. PATIENTS AND METHODS: All children undergoing anatrophic nephrolithotomy for complex branching and multiple renal calculi over an 11-year period were studied prospectively. Demographic data, treatment details and outcome, as assessed by X-ray, ultrasonography and isotope studies, were recorded. Anatrophic nephrolithotomy was carried out with surface cooling of the kidney followed by nephrostomy drainage for 5-7 days. RESULTS: Nine children (median age 4 years, range 7 months to 9 years) underwent anatrophic nephrolithotomy. Predisposing factors included urinary tract infection (by Proteus mirabilis) in all and hyper-calciuria in two children. The median (range) total ischaemic time at operation was 25 (15-40) min and the operative duration 150 (120-200) min. Three children required a blood transfusion. Stone clearance was incomplete in one child. There was no recurrent stone formation after a long term follow-up (median 32 months, range 14-107) in the other patients. Isotope studies showed impaired split renal function (<40%) in six children before surgery; there was a significant decline (>5%) in divided function in five children (range 6-16%) after surgery. CONCLUSION: Anatrophic nephrolithotomy is an effective means of rendering children with branching calculi stone-free, but this study suggests that it leads to some further parenchymal damage. PMID- 10792170 TI - Bladder calculi in children who perform clean intermittent catheterization. AB - OBJECTIVE: To examine the role of clean intermittent catheterization (CIC) as a possible predisposing risk factor for bladder calculi, assessing risk factors in patients with and without bladder augmentation, and to evaluate management options for bladder calculi in these patients. PATIENTS AND METHODS: The records of 403 patients who were using a regimen of CIC between January 1981 and March 1998 were reviewed to identify those forming bladder calculi; stones were diagnosed in 28 patients. The patients were categorized as: group 1, patients with no bladder augmentation who catheterized urethrally (227, group 1a) or via a Mitrofanoff conduit (18, group 1b); group 2, patients with augmented bladders who catheterized urethrally (100, group 2a) or via a Mitrofanoff conduit (58, group 2b). The incidence of bladder calculi in each group was determined and compared statistically where applicable. The success of the treatment options for stone management was reviewed. RESULTS: Bladder calculi developed in 5% of patients in group 1a, 8% in group 2a, 11% in group 1b, and 10% in group 2b; the incidence of calculi was not significantly different among the groups. Of these patients, 18 (64%) were asymptomatic at the time of diagnosis and significant bacteriuria was found in 23 (88%). Difficulty in catheterizing either the Mitrofanoff conduit or the native urethra was reported in 14 (50%) of these patients. Calculi were more often solitary (71%) and typically composed of struvite or apatite. Calculi were managed by open cystolithotomy in 15 patients (54%) and endoscopically in 13 (46%). Stones recurred in nine patients (32%) after treatment, comprising four of six patients treated endoscopically with electrohydraulic lithotripsy and in five of 15 after open cystolithotomy. The mean interval to recurrence was 22.8 months. CONCLUSION: These results suggest that patients on a regimen of CIC are at risk of developing bladder calculi but the incidence of calculi is not influenced by bladder augmentation. The presence of a Mitrofanoff conduit was associated with a slightly increased incidence of calculus formation. Open cystolithotomy was associated with a lower stone recurrence rate but there were too few patients to draw definitive conclusions. PMID- 10792171 TI - Urinary tract abnormalities in children with acute focal bacterial nephritis. AB - OBJECTIVE: To review the presentation, methods of diagnosis, and incidence of concomitant urological abnormalities in children with acute focal bacterial nephritis (AFBN, also known as lobar nephronia, a severe nonliquefactive infection involving one or more renal lobules). PATIENTS AND METHODS: Sixteen children (age range 6 months to 9 years) diagnosed with AFBN over a 5-year period were retrospectively reviewed. Their age, gender, presenting symptoms, presence of urinary tract abnormalities, radiological imaging and treatment were assessed. RESULTS: All 16 children received intensive intravenous and oral antibiotics. Ten required intervention for predisposing conditions including vesico-ureteric reflux (four), bladder diverticulum (one), renal and peri-rectal abscess (two), ureteric or bladder calculi (two), and renal mass (one). In addition, three children had reflux not requiring surgery; one had multiple renal calyceal diverticula, one had Hinman syndrome and one had leukaemia. CONCLUSION: Acute focal bacterial nephritis is being diagnosed with increasing frequency through increasing awareness and advances in imaging modalities. Active urological evaluation is warranted because of the high incidence of urinary tract abnormalities. In addition to antibiotics, intervention was needed in most children in this series. PMID- 10792172 TI - Pelvic-floor therapy and toilet training in young children with dysfunctional voiding and obstipation. AB - OBJECTIVE: To analyse experience in treating young children (4-5 years old) with urodynamically confirmed voiding dysfunction, using a noninvasive training programme. PATIENTS AND METHODS: Between January 1996 and October 1997, 20 children (all < 5 years old, mean age 4.45 years, 18 girls and two boys, mean ages 4.44 and 4.5 years, respectively) with voiding dysfunction were treated. Three children showed filling phase dysfunction alone (bladder instability), six emptying phase dysfunction alone (dysfunctional voiding) and 11 showed both filling and emptying phase dysfunction. Sixteen children had incontinence problems (three diurnal, two nocturnal and 11 diurnal and nocturnal). Eight children had a history of recurrent urinary tract infections and 12 girls had vaginal irritation. Four children were referred for perineal pain caused by spasms of the pelvic floor. Eight children had encopresis based on chronic obstipation. Therapy consisted of keeping a voiding and drinking chart, instructions on proper toilet posture, daily rules for application at home, and if possible relaxation biofeedback of the pelvic-floor muscles. Therapy was considered successful if incontinence and other urological symptoms resolved. The treatment of encopresis is also discussed. RESULTS: Of the 20 children, 13 had a good result; they all became dry during the day and night, and encopresis resolved. Six children had moderate success; in one, nocturnal incontinence persisted, and in two diurnal and nocturnal incontinence continued. In two children encopresis persisted and in one the faecal incontinence ameliorated. In one child the therapy was prematurely interrupted because of lack of motivation. CONCLUSION: This experience suggests that a noninvasive training programme is applicable in very young children with symptoms of dysfunctional elimination of urine and faeces. PMID- 10792173 TI - Acellular bladder matrix allografts in the regeneration of functional bladders: evaluation of large-segment (> 24 cm) substitution in a porcine model. AB - OBJECTIVES: To evaluate the use of a large-segment (> 24 cm2) bladder substitution with porcine bladder acellular matrix allograft (BAMA) in a large animal model. Materials and methods Bladders were harvested from pigs at the time of necropsy and subjected to detergent and enzymatic extractions to render them acellular. The BAMA produced had the surgical handling and suture-retaining properties of normal bladder tissue. Six pigs had BAMA segments implanted under general anaesthesia, through a low midline abdominal incision and after partial cystectomy. The defect was repaired with a BAMA patch (mean size 43.88 cm2, range 12-72), with no urinary diversion. Two animals each were then killed at 9, 16 and 30 days and the bladders explanted. The native bladder and BAMA patch were analysed morphometrically to evaluate cellular re-population and matrix re organization. RESULTS: All animals survived surgery; there were no urinary leaks and no stones detected in any of the bladders. At 9 days there was a diffuse infiltration with acute inflammatory cells, but no areas of necrosis. There were isolated areas of smooth muscle cell (SMC) infiltration of the BAMA. At 16 days the luminal surface was lined with a single layer of urothelium, there was stromal infiltration with disorganized SMC and angiogenesis, with mature vessels in the BAMA patch. At 30 days the urothelium was multilayered with organizing groups of SMCs and angiogenesis. The highest cell density was at the periphery of the repopulated BAMA patch, decreasing towards the centre. CONCLUSIONS: The implantation of large patches of BAMA is technically feasible and may prove to be a viable surgical alternative to bladder augmentation with intestinal segments. The advantages of BAMA include the potential for complete and functional regeneration of a bladder substitute. This model provides a tool with which to obtain a better understanding of the cellular and molecular aspects of matrix re population. PMID- 10792174 TI - Effect of age on the responses of rat bladder detrusor strips to adenosine triphosphate. AB - OBJECTIVE: To assess age-related changes in bladder function using the contractile responses to ATP of detrusor strips from rats of various ages. Materials and methods Urinary bladders were obtained from male Wistar rats aged 9 weeks (young), 24 weeks (adult) and 24 months (aged). Contractions of urinary bladder muscle strips to ATP were measured isometrically. The size of the initial phasic response and the secondary contractile response that developed after washing out ATP (postwashout contraction) were measured. The magnitudes of the ATP-induced phasic and postwashout contraction were compared among the age groups. During the contractions, prostanoid concentrations in the organ-bath medium were measured using an enzyme immunoassay. RESULTS: The ATP-induced postwashout contraction did not occur after stimulation with KCl or acetylcholine, but was induced by alpha,beta-methylene ATP. Both the phasic and postwashout contractions were concentration-dependent. Although the phasic contraction did not change progressively with age, the magnitude and duration of the postwashout contraction increased substantially with age. Nicardipine (a calcium antagonist) slightly inhibited both contractions. Suramin (a nonselective P2-receptor antagonist) did not significantly inhibit the phasic contraction, but reduced the postwashout contraction. PPADS (a selective P2X receptor antagonist) did not inhibit either contraction. Indomethacin (a prostaglandin synthesis inhibitor) had no effect on the phasic contraction but almost completely blocked the postwashout contraction when added before ATP stimulation, but was less effective when added after ATP. The prostaglandin E2 concentration in the organ bath increased during the postwashout contraction. CONCLUSIONS: These findings suggest that the ATP-induced postwashout contraction is not directly mediated by P2x purinoceptors, but results from the synthesis of prostaglandins, especially E2, which is a sensory autacoid. The age-linked increase in postwashout contraction may be involved in the changes in sensory and voiding mechanisms seen in the aged urinary bladder. PMID- 10792175 TI - Role of C-afferent fibres in the mechanism of action of sacral nerve root neuromodulation in chronic spinal cord injury. AB - OBJECTIVE: To determine whether sacral root neuro-modulation (a promising therapeutic modality in patients with refractory voiding and storage problems) has its effect through the blockade of C-afferent fibres that form the afferent limb of a pathological reflex arc responsible for the dysfunction of bladder storage. MATERIALS AND METHODS: The study comprised 39 female Sprague Dawley rats divided into three equal groups: normal controls (group 1); spinally transected at T10 (group 2); spinally transected and electrically stimulated bilaterally at S1 for 6 h daily (group 3). Three weeks after transection the rats were assessed using urodynamics; substance P, neurokinin A and calcitonin gene-related peptide (CGRP) were extracted from the dorsal root ganglia (DRG) of the L5 and L6 roots and quantified by radioimmunoassay. RESULTS: Spinally transected rats developed urinary bladder hyper-reflexia after 3 weeks. This was associated with a significant increase in the neuropeptide content of the DRG of L6. Electrostimulation of S1 significantly decreased the neuropeptide content of L6. In contrast, transection and S1 neurostimulation did not affect the neuropeptide content of the L5 DRG, except for CGRP, which increased after spinal transection and decreased with neurostimulation. CONCLUSIONS: In spinally transected rats, sacral root neurostimulation abolished bladder hyper-reflexia and attenuated the rise in neuropeptide content of the L6 DRG. These results suggest that the blockade of C-afferent fibre activity is one of the mechanisms of action of sacral root neuromodulation. PMID- 10792176 TI - Induction of apoptosis by mitomycin-C in an ex vivo model of bladder cancer. AB - OBJECTIVE: To examine mitomycin-C (MMC)-induced apoptosis in an ex vivo model of superficial TCC, and relate it to the in vivo response to chemotherapy. Materials and methods Dose- and time-response curves were constructed to determine the optimal conditions for the induction of apoptosis by MMC in an ex vivo model of superficial bladder cancer. Subsequently, 41 individual tumours were exposed to MMC in the model and the effects assessed by measuring of apoptosis before and after chemotherapy. The relationships between tumour grade and stage and the intrinsic and induced apoptotic counts were determined. In tandem, in a clinical study, the relationship between in vivo response of a marker tumour to MMC and the ex vivo induction of apoptosis was determined. RESULTS: In the ex vivo model, apoptosis was induced at a MMC concentration of 0.5 mg/mL after an incubation time of 8 h. In 41 tumours the intrinsic apoptotic index (AI) was higher with increased grade and stage of tumour (P = 0.048). There was no correlation between the intrinsic AI and the AI after treatment with MMC (induced AI). In 21 tumours (51%) the induced AI did not increase above a predetermined response threshold and these tumours were considered resistant to MMC. Resistance to MMC was related to tumour grade (P = 0.037) with a trend for G3 pT1 tumours to be resistant to the therapy. There was a significant association between ex vivo sensitivity and in vivo marker tumour response (P = 0.02). CONCLUSIONS: Apoptosis is differentially induced in an ex vivo incubation model of superficial TCC by MMC and evidence suggests that this response matches that seen in vivo. The measurement of apoptosis before therapy does not predict the apoptotic response of a tumour to chemotherapy. The ability to undergo apoptosis correlates with clinical outcome. PMID- 10792177 TI - Lack of neuropeptide Y receptor detection in human bladder and prostate. AB - OBJECTIVE: To investigate the presence of functional neuropeptide Y (NPY) receptors in human bladder and prostate (both richly endowed with NPY-containing nerve fibers) using peptide YY (PYY) as the agonist. Materials and methods Binding studies were conducted using [125I]PYY as the radioligand. Organ-bath studies were performed on isolated tissue strips for direct (postjunctional) contractile effects and for (prejunctional) inhibition of field stimulation effects. Any possible degradation of PYY was determined using high-performance liquid chromatography (HPLC). RESULTS: In the radioligand binding studies no quantifiable specific [125I]PYY binding was detected in human bladder or prostate, while specific high-affinity binding was readily seen in rat cerebral cortex. In organ-bath experiments, PYY (up to 1 micromol/L) caused no contraction of human prostate or bladder, whereas noradrenaline and carbachol, respectively, were effective; the potency or efficacy of noradrenaline and carbachol were not altered by PYY. Field stimulation-induced contraction was not affected by PYY in either human bladder or prostate, but was readily inhibited in rat vas deferens. HPLC detected no relevant PYY degradation by human bladder or prostate homogenates. CONCLUSION: Human bladder and prostate express only very few if any functional NPY receptors. PMID- 10792178 TI - The role of spinal and peripheral alpha1- and alpha2-adrenoceptors on bladder activity induced by bladder distension in anaesthetized rat. AB - OBJECTIVE: To elucidate the effects of alpha1- and alpha2-adrenoceptor antagonists on the micturition reflex induced by bladder distension in the anaesthetized rat at the peripheral and spinal level. Materials and methods Using continuous cystometry in 80 anaesthetized female Sprague-Dawley rats, changes in basal pressure (BaP), micturition pressure (MP), bladder capacity (BC), micturition volume (MV), interval of bladder contraction (frequency) and residual volume (RV) to the selective intra-arterial and intrathecal administration of drugs were monitored and analysed. RESULTS: The intra-arterial administration of phentolamine, prazosin, doxazosin and tamsulosin significantly decreased the MP; doxazosin increased MV, BC, RV and frequency. MP was inhibited more by intra arterial doxazosin than by prazosin or tamsulosin. Intrathecal phentolamine increased BaP, BC, RV and decreased MP and MV; tamsulosin decreased frequency and increased BC. Clonidine markedly increased BaP and frequency, whereas MV and BC were significantly lower; yohimbine increased MV, BC and decreased RV. Intra arterial yohimbine significantly increased MV. CONCLUSIONS: At the spinal level, the micturition reflex evoked by bladder distension was preferentially suppressed by phentolamine or yohimbine, suggesting the involvement of alpha2-adrenoceptors. In the periphery, alpha1-adrenoceptor blockers preferentially suppressed the reflex. We suggest that both alpha1- and alpha2-adrenoceptors play a role in the distension-evoked micturition reflex in the rat. PMID- 10792179 TI - Phenotypic and genotypic characterization of commonly used human prostatic cell lines. AB - OBJECTIVE: To investigate and catalogue systematically the phenotypic and genotypic characteristics of the commonly used prostatic cell lines using immunocytochemistry and polymerase chain reaction (PCR) of hypervariable sequences within the genome to provide a 'fingerprint' characteristic of each cell line. Materials and methods Malignant (LNCaP, LNCaP-r, PC-3, DU-145) and benign immortalized prostatic cell lines (PNT-1A, PNT-1B, BPH-1) were grown on four-well slides, fixed and subjected to indirect streptavidin-biotin immunocytochemistry. Twenty-three antibodies were used in the following groups: cytoskeletal elements: cytokeratins (CK)-5, -7, -8, -14 (two), -16, -18, -19 (three), -20, vimentin and desmin; MUC1 (three); cell adhesion molecules (E cadherin, alpha-beta-and gamma-catenin); and prostatic associated proteins: prostate specific antigen (PSA), prostatic acid phosphatase (PAP) and androgen receptor (AR). For the PCR, genomic DNA was extracted from the cell lines and from SKOV3 and MCF7 (positive controls). PCR was performed on three variable regions which were then sequenced: AR exon 1 (CAG repeat polymorphism), and two areas of microsatellite instability (MSI): AR exon 8 and hypoxanthine-guanine phosphoribosyl transferase (HPRT) exon 3. RESULTS: All cell lines were CK-8/18 positive and most also expressed CK-7 and -19. Heterogeneous CK-20 expression was detected for the first time in prostatic cell lines. All lines were positive for vimentin and negative for desmin. MUC1 was expressed in one malignant (DU-145) and all immortalized cell lines. E-cadherin expression was low or absent in three lines: PNT1A, 1B and PC-3. Only PC-3 failed to express alpha-catenin; beta- and gamma-catenin were expressed by all lines. PSA, PAP and AR were only expressed by LNCaP and LNCaP-r. On PCR, the CAG repeat lengths in exon 1 of the AR ranged from 19 to 27. Three pairs of cell lines had the same exon 1 CAG repeat length: LNCaP/PC-3 (26 repeats), BPH-1/DU-145 (19 repeats) and PNT1 A/1B (20 repeats). Exon 8 sequences were identical except for LNCaP, which showed a single base mutation, and HPRT exon 3 sequences were all identical. There was no evidence of generalized MSI in any of the cell lines examined. CONCLUSIONS: The cell lines studied fell into three broad groups according to their phenotypic characteristics: (i) prostatic marker positive (LNCaP and LNCaP-r); (ii) high expression of most antigens (DU-145, PC-3 and BPH-1); and (iii) low or absent expression of most antigens (PNT1 A and 1B). Each of the cell lines derived from PC could be identified on the basis of exon 1 and 8 AR sequence variability. DU145 and BPH-1 had identical profiles of the three areas studied, but these cell lines are easily distinguished by their different phenotypic characteristics. PNT1A and 1B had identical genetic and similar phenotypic profiles, which is unsurprising given that they are subclones derived from the same parental line. Even so, these were separable on the basis of CK-19 immunostaining. Using a combination of geno- and phenotypic markers it was possible to derive a 'fingerprint' for each of the cell lines assessed, which will allow meaningful comparison between similar cell lines held in other laboratories. PMID- 10792181 TI - Delayed testosterone replacement restores nitric oxide synthase-containing nerve fibres and the erectile response in rat penis. AB - OBJECTIVE: To elucidate the effect of testosterone on penile innervation. Materials and methods Three groups of six rats each were assessed; two groups (1 and 2) were castrated and the third (group 3) underwent a sham operation (control). Eight weeks after castration, group 2 received a subcutaneous injection with testosterone. At 8 weeks, the rats in group 1 and 3 underwent a final functional analysis while those in group 2 did so at 12 weeks. The evaluation included a subcutaneous injection with apomorphine to study centrally mediated erection, and cavernosal nerve electrostimulation and papaverine injection to study peripherally mediated erection. At death a penile mid-shaft specimen was taken for NADPH-diaphorase staining. RESULTS: In the apomorphine study, castration resulted in significantly fewer yawns and erections than in the control, and those in group 2 significantly better central erectile function than in the controls. The mean (SEM) number of nitric oxide synthase (NOS)-containing nerve fibres in the corpora cavernosa and both dorsal nerves of castrated rats, at 46.2 (9.1) and 203 (32.1), respectively, were significantly lower than in rats in group 2, at 84.1 (11.2) and 300.6 (17.1), and than in the controls, at 88.6 (10.9) and 306.3 (22.9), respectively. The intracavernosal pressure decreased significantly in the absence of testosterone, both after electrostimulation and intracavernosal papaverine injection. However, there was no difference between the control and group 2 rats in either the number of NOS-containing nerve fibres or in the peripheral erectile functional study. CONCLUSIONS: Testosterone acts on the nervous system to mediate erection; when it is absent there may be down regulation of both the production and activity of NO, thereby decreasing the response to peripheral stimulation via the NO pathway. The restoration of erectile function seen in rats in group 2 supports this phenomenon. Delayed testosterone replacement has no detrimental effect on the restoration of the erectile mechanism after castration. PMID- 10792180 TI - Expression and localization of basic fibroblast growth factor in diabetic rat prostate. AB - OBJECTIVE: To assess fibroblast growth factor-2 (FGF2/bFGF), which is important in the development and maintenance of the normal prostate and in the development of human benign prostatic hyperplasia (BPH) and prostatic carcinoma, in an animal model of experimentally induced diabetes. Materials and methods Using Western blotting and immunohistochemical analyses, the expression of FGF2 in prostates from several groups of rats was investigated. Rats had diabetes for 8 or 16 weeks (induced by intravenous injection with 65 mg/kg streptozotocin); rats were also treated with insulin (starting 8 weeks after the induction of diabetes, for 8 weeks), and two further groups acted as age-matched control rats. Immunohistochemical markers for smooth muscle (alpha-actin) and epithelium (cytokeratin) were used to distinguish different cell types in adjacent prostatic sections. RESULTS: Diabetic rats had smaller prostates and lower serum testosterone levels than their controls; insulin treatment of diabetic rats increased prostatic size and testosterone levels. As shown by Western blotting, diabetes caused greater FGF2 expression than in controls, whereas reverse transcriptase polymerase chain reaction studies showed similar levels of prostatic FGF-2 mRNA in all groups. Immuno-histochemical studies showed that FGF 2 was expressed in both stromal and epithelial components of the rat prostate. Furthermore, although the expression of FGF2 was higher in epithelial than stromal cells in control prostates, it was distributed uniformly in the diabetic prostate. CONCLUSION: The differences in the level of expression and pattern of distribution of FGF2 suggests a potential role for FGF2 in the changes observed in prostatic growth in diabetic rats. PMID- 10792182 TI - Affinity of trazodone for human penile alpha1- andalpha2-adrenoceptors. AB - OBJECTIVE: To determine the affinities of human penile alpha-adrenoceptors and cloned human alpha-adrenoceptor subtypes for trazodone, an antidepressant with reported beneficial effects in erectile dysfunction. Materials and methods Competition radioligand binding studies were performed with trazodone in human penile tissue and in cell lines stably expressing all cloned human alpha adrenoceptor subtypes. RESULTS: Trazodone had higher affinity for human alpha1 adrenoceptors (10-130 nmol/L) than for alpha2-adrenoceptors (300-700 nmol/L), but did not discriminate between subtypes of human alpha1- or alpha2-adrenoceptors. CONCLUSION: Trazodone has high and moderate affinity for human alpha1- and alpha2 adrenoceptors, respectively; this might contribute to its reported beneficial effects in erectile dysfunction. PMID- 10792184 TI - Transurethral circum-apical incision of the prostate: minimizing sphinteric injury during transvesical prostatectomy for large prostates. PMID- 10792183 TI - Effects of finasteride and bicalutamide on prostatic blood flow in the rat. AB - OBJECTIVE: To determine whether finasteride and bicalutamide, both currently used in the clinical management of patients with prostate diseases because they have anti-androgenic properties, have any effects on prostatic blood flow in a rat prostate model, as androgens are known to be involved in the regulation of prostatic blood flow and angiogenesis. MATERIALS AND METHODS: Both finasteride and bicalutamide were supplied as oral suspensions in water and given daily to rats for 7 days by tube feeding. Blood flows to the ventral and dorsal prostates, and to the kidneys, were measured using the radioactive microsphere technique. In the bicalutamide experiments, some rats were treated with the Leydig cell toxin ethane dimethane sulphonate (EDS), to obtain a castration-like effect, and one group of these rats received testosterone. RESULTS: Finasteride induced a clear decrease in blood flow to the ventral and dorsal prostates after 7 days of treatment, with no significant changes in blood pressure or kidney blood flow. Bicalutamide inhibited the testosterone-induced increment of prostatic blood flow observed in EDS-treated animals. CONCLUSIONS: Finasteride, a blocker of 5alpha reductase, decreases prostate blood flow after 7 days of administration. The response was slower than that after castration, but was of similar magnitude. Blood flow was also decreased after treatment with the androgen-receptor inhibitor bicalutamide. These observations suggest that prostatic blood flow is increased by dihydrotestosterone, and that the androgen receptor is responsible for mediating this effect. PMID- 10792185 TI - A renal foreign-body stone as a complication of myelography. PMID- 10792186 TI - An unusual cause of testicular ischaemia. PMID- 10792188 TI - Reply PMID- 10792187 TI - Antenatally detected pelvi-ureteric junction obstruction: concerns about conservative management. PMID- 10792189 TI - Penile refracture. PMID- 10792190 TI - Penile refracture. PMID- 10792191 TI - The urology of Pharaonic Egypt. PMID- 10792192 TI - Renal transplantation in children. PMID- 10792193 TI - Antenatal hydronephrosis: changing concepts in diagnosis and subsequent management. PMID- 10792194 TI - Interstitial cystitis: a review of immunological aspects of the aetiology and pathogenesis, with a hypothesis. PMID- 10792195 TI - Ureteric reimplantation: a history of the development of surgical techniques. PMID- 10792196 TI - Within- and between-subject variations in pharmacokinetic parameters of ethanol by analysis of breath, venous blood and urine. AB - AIMS: To evaluate the prerequisites for using ethanol dilution to estimate total body water, we studied the within- and between-subject variation in the parameter estimates of a two-compartment model for ethanol pharmacokinetics with parallel Michaelis-Menten and first-order renal elimination. Because sampling of breath might be preferable in some clinical situations the parameter estimates derived from breath and venous blood were compared. METHODS: On two occasions, ethanol 0.4 g kg-1 was given by intravenous infusion to 16 volunteers after they had fasted overnight. The proposed model was fitted by means of nonlinear regression to concentration-time data measured in the breath, venous blood and urine during 360 min. The model contained six parameters: Vmax and Km (Michaelis-Menten elimination constants), CLd (intercompartmental distribution parameter), VC and VT (volumes of the central and tissue compartment, respectively) and CLR (renal clearance). The volume of distribution, Vss, was calculated as the sum of VC and VT. RESULTS: The mean +/- total s.d. of the parameter estimates derived from blood data were Vmax 95 +/- 25 mg min-1, Km 27 +/- 19 mg l-1, CLd 809 +/- 232 ml min-1, VC 14.5 +/- 4.3 l, VT 21. 2 +/- 4.4 l, CLR 3.6 +/- 2.0 ml min-1 and Vss 35.8 +/- 4.3 l. The variation within subjects amounted to 3%, 9%, 21%, 21%, 17%, 26% and 2%, respectively, of the total variation. Breath samples were associated with a similar or lower variation than blood, both within and between subjects. About 1.5% of the infused ethanol was recovered in the urine. CONCLUSIONS: The low within-subject variation of the key parameter Vss (only 2%) suggests that ethanol dilution analysed by the pharmacokinetic model applied here may be used as an index of the total body water. Breath samples yielded at least as good reproducibility in the model parameters as venous blood. PMID- 10792197 TI - Apparent autoinduction of valproate beta-oxidation in humans. AB - AIMS: The study aimed to show whether autoinduction of valproate (VPA) along its beta-oxidation pathway occurred upon chronic dosing in humans. METHODS: Twelve young volunteers without active illness took sodium valproate (NaVPA) 200 mg orally 12 hourly for 3 weeks. On days 7 and 21, serial blood samples and all urine passed over an interdosing interval from 08.00 to 20.00 h were collected for analysis of VPA and certain metabolites. RESULTS: Plasma AUC(0,12 h) of VPA was significantly lower on day 21 than on day 7 (2.40 vs 2.84 micromol ml-1 h, 95% CI for the difference 0.13-0.81 micromol ml-1 h). Significant differences in plasma AUC(0,12 h) of the beta-oxidation metabolites E-2-en-VPA and 3-oxo-VPA were not found. However, formation clearances of plasma VPA to urinary E-2-en-VPA and 3-oxo-VPA were significantly increased from day 7 to day 21 (0. 010 vs 0.024 and 2.57 vs 3.60 ml kg-1 h-1, respectively, 95% CI for the differences -0.025 to 0.004 and -1.72 to -0.34 ml kg-1 h-1, respectively). Formation clearances to VPA glucuronide (0.534 vs 0. 505 ml kg-1 h-1) and 4-OH-VPA (0.112 vs 0.110 ml kg-1 h 1) were not significantly different. CONCLUSIONS: Regular low dose VPA intake in humans over a period of 3 weeks appears to be associated with a small induction of its metabolism by the beta-oxidation pathway, but not by glucuronidation or 4 hydroxylation. PMID- 10792198 TI - The single dose pharmacokinetics of ribavirin in subjects with chronic liver disease. AB - AIMS: The primary objective of this study was to describe the single dose pharmacokinetics of ribavirin in subjects with normal liver function and those with various degrees of stable chronic liver disease. Additionally this study assessed the safety and tolerability of ribavirin in this population. METHODS: Single oral 600 mg doses of ribavirin were administered to healthy male and female volunteers (n = 6) and patients with stable chronic liver disease (n = 17), in a parallel group study. Pharmacokinetic sampling and tolerability assessments were performed up to 168 h post dose. RESULTS: Single oral doses of 600 mg ribavirin were well tolerated by healthy volunteers and patients with varying degrees of hepatic dysfunction. Although mean Cmax increased with the severity of hepatic dysfunction, there was no change in extent of absorption or renal clearance of ribavirin. CONCLUSIONS: There are no pharmacokinetic reasons for initial dose adjustment of ribavirin in patients with hepatic dysfunction. PMID- 10792199 TI - Cardiovascular effects of melatonin in hypertensive patients well controlled by nifedipine: a 24-hour study. AB - AIMS: As melatonin has been found to play a role in the mechanisms of cardiovascular regulation, we designed the present study to evaluate whether the evening ingestion of the pineal hormone might interfere with the antihypertensive therapy in hypertensive patients well-controlled by nifedipine monotherapy. METHODS: Forty-seven mild to moderate essential hypertensive outpatients taking nifedipine GITS 30 or 60 mg monotherapy at 08.30 h for at least 3 months, were given placebo or melatonin 5 mg at 22.30 h for 4 weeks according to a double blind cross-over study. At the end of each treatment period patients underwent a 24 h noninvasive ambulatory blood pressure monitoring (ABPM) during usual working days; sleeping period was scheduled to last from 23.00 to 07.00 h. RESULTS: The evening administration of melatonin induced an increase of blood pressure and heart rate throughout the 24 h period (DeltaSBP = + 6.5 mmHg, P < 0.001; DeltaDBP = + 4.9 mmHg, P < 0.01; DeltaHR = + 3.9 beats min-1, P < 0.01). The DBP as well as the HR increase were particularly evident during the morning and the afternoon hours. CONCLUSIONS: We hypothesize that competition between melatonin and nifedipine, is able to impair the antihypertensive efficacy of the calcium channel blocker. This suggests caution in uncontrolled use of melatonin in hypertensive patients. As the pineal hormone might interfere with calcium channel blocker therapy, it cannot be considered simply a dietary supplement. PMID- 10792200 TI - Effects of the 5-HT3 antagonist cilansetron vs placebo on phasic sigmoid colonic motility in healthy man: a double-blind crossover trial. AB - AIMS: 5-hydroxytryptamine3 receptor antagonists act antiemetically and slow colonic transit. This study evaluated effects of the high-affinity 5-HT3 antagonist, cilansetron, on fasting, meal-and anticholinesterase-stimulated phasic contractile activity of the human sigmoid colon as well as on bowel habits and stool consistency. METHODS: Five female and seven male healthy volunteers received, during three 7 day periods separated by 7 day wash-out periods, 4 mg cilansetron, 8 mg cilansetron or placebo three times daily orally under random, double-blind, crossover conditions. On day 8 of each treatment period, motility 20-40 cm from the anal verge was recorded using five pressure sensors spaced at 5 cm intervals. After a basal 30 min, subjects swallowed a further dose of the scheduled treatment; 60 min later, blood was taken for the determination of plasma cilansetron levels. Thereafter, subjects ingested a 4200 kJ meal and 250 ml sweetened mallow tea (166 kJ); 90 min after meal onset, 1 mg neostigmine was administered intramuscularly and motility recording was continued for 60 min RESULTS: Phasic contractile activity and intraluminal base-line pressure increased postprandially and more so after neostigmine. With cilansetron, the area under the pressure curve as the primary outcome variable and the number of contractions were significantly greater than with placebo (P = 0.005), amplitude and duration of contractions and base-line pressure were not affected. The effects of the two cilansetron dosages did not differ. With cilansetron, stool tended to become firmer. No adverse effects were observed. Plasma levels were highest with 8 mg cilansetron. CONCLUSIONS: Cilansetron slightly augments meal stimulated and markedly neostigmine-stimulated phasic motility of the sigmoid colon. When administered over 7 days, it tends to increase stool consistency and is well tolerated. PMID- 10792201 TI - In vitro sensitivity of Plasmodium falciparum and clinical response to lumefantrine (benflumetol) and artemether. AB - AIMS: To assess the sensitivity of 103 Plasmodium falciparum isolates to a combination of lumefantrine (benflumetol) and artemether (CGP 56697), with the objective of determining a correlation between in vitro drug sensitivity and therapeutic outcome. METHODS: Patients suffered from uncomplicated falciparum malaria and came from areas of Thailand affected by multidrug resistance. CGP 56697 was given in the form of tablets containing 20 mg artemether and 120 mg lumefantrine. The standard dose regimen, 4 doses of 4 tablets over 48 h, was compared with two lower dose regimens (4 x 2 tablets and 3 x 4 tablets). RESULTS: The parasites showed high resistance to chloroquine, fairly advanced resistance to mefloquine and compromised sensitivity to quinine. Sensitivity to artemisinin and lumefantrine prior to treatment was similar in all treatment groups. The 4 x 4 tablet regimen was more effective than the other regimens in coping with infections with relatively low sensitivity to artemisinin and/or lumefantrine. The EC90 for artemisinin is an important determinant of treatment success. Parasite density at the start of treatment was identified as another critical predictor of treatment outcome. CONCLUSIONS: The results indicate that parasite exposure to the drugs may have been inadequate and/or too short in the cases of treatment failure, particularly marked in the lower dose regimens. This could probably be remedied by expanding the dose regimen in areas affected by multidrug resistance and in the case of relatively high parasitaemia. PMID- 10792204 TI - Lack of adherence to lipid-lowering drug treatment. A comparison of utilization patterns in defined populations in Funen, Denmark and Bologna, Italy. AB - AIMS: The objective was to explore differences in lipid-lowering drug (LLD) prescribing in Italy and Denmark. METHODS: We used two geographical areas with computerized drug prescription records in defined populations, one in Funen, Denmark with 500 000 inhabitants, the other in Bologna, Italy with 400 000 inhabitants. Prescriptions for patients who had purchased a LLD from 1994 until 1996 were retrieved as well as coprescriptions of antidiabetic and cardiovascular drugs as markers for diabetes and cardiovascular disease. Only patients surviving and remaining in the area were included. Compliance was defined as percentage of DDDs purchased divided by the number of days within the time window. The limit between good and poor compliance was set at 82%. RESULTS: In Bologna, LLD consumption measured in DDD increased by 41% and in Funen by 129%. Annual prevalence increased from 36.9 to 46.3 users/1000 inhabitants from 1994 to 1996 and from 3.2 to 6.6 users/1000 inhabitants in Bologna and Funen, respectively. From 1995 to 1996, the incidence of use decreased slightly in Bologna from 19.3 to 18. 8/1000 inhabitants/year, whereas in Funen the incidence increased from 1.8 to 2.3/1000 inhabitants/year. In Bologna 48% and in Funen 91% of users persisted with treatment for 2 years or longer. In Bologna, 7% and in Funen 45% were good compliers. In Bologna, 61% and in Funen, 72% received other drugs indicating cardiovascular or diabetic comorbidity. CONCLUSIONS: Patterns of use differed substantially between the two areas. In contrast with Funen, where long-term use was common, Bologna LLD use was sporadic. Based on a higher rate of coprescription, LLDs seemed to be used for secondary prevention to a higher extent in Funen than in Bologna. In Funen it appeared that the correct patients, but an insufficient number of them, were being treated adequately according to guidelines. The higher discontinuation rate of lipid lowering drugs in the Bologna area indicates that a large proportion of patients use these drugs for too short a period of time to benefit from treatment. Since society's health care resources are limited it is difficult to justify public funding of these medications without at the same time giving appropriate attention to these problems. PMID- 10792202 TI - Pharmacokinetic-pharmacodynamic evaluation of ceftazidime continuous infusion vs intermittent bolus injection in septicaemic melioidosis. AB - AIMS: Experimental studies have suggested that constant intravenous infusion would be preferable to conventional intermittent bolus administration of beta lactam antibiotics for serious Gram-negative infections. Severe melioidosis (Burkholderia pseudomallei infection) carries a mortality of 40% despite treatment with high dose ceftazidime. The aim of this study was to measure the pharmacokinetic and pharmacodynamic effects of continuous infusion of ceftazidime vs intermittent bolus dosing in septicaemic melioidosis. METHODS: Patients with suspected septicaemic melioidosis were randomised to receive ceftazidime 40 mg kg 1 8 hourly by bolus injection or 4 mg kg-1 h-1 by constant infusion following a 12 mg kg-1 priming dose to perform estimation of pharmacokinetic and pharmacodynamic parameters. RESULTS: Of the 34 patients studied 16 (59%) died. Twenty patients had cultures positive for B. pseudomallei of whom 12 (60%) died. The median MIC90 of B. pseudomallei was 2 mg l-1, giving a target concentration CT, of 8 mg l-1. The median (range) estimated total apparent volume of distribution, systemic clearance and terminal elimination half-lives of ceftazidime were 0.468 (0.241-0.573) l kg-1, 0.058 (0.005-0.159) l kg-1 h-1 and 7.74 (1.95-44.71) h, respectively. Clearance of ceftazidime and creatinine clearance were correlated closely (r = 0. 71; P < 0.001) and there was no evidence of significant nonrenal clearance. CONCLUSIONS: Simulations based on these data and the ceftazidime sensitivity of the B. pseudomallei isolates indicated that administration by constant infusion would allow significant dose reduction and cost saving. With conventional 8 h intermittent dosing to patients with normal renal function, plasma ceftazidime concentrations could fall below the target concentration but this would be unlikely with a constant infusion. Correction for renal failure which is common in these patients is Clearance = k * creatinine clearance where k = 0.072. Calculation of a loading dose gives median (range) values of loading dose, DL of 3.7 mg kg-1 (1. 9-4.6) and infusion rate I = 0.46 mg kg h-1 (0.04-1.3) (which equals 14.8 mg kg-1 day-1). A nomogram for adjustment in renal failure is given. PMID- 10792203 TI - Nonspecific binding of drugs to human liver microsomes. AB - AIMS: To characterize the nonspecific binding to human liver microsomes of drugs with varying physicochemical characteristics, and to develop a model for the effect of nonspecific binding on the in vitro kinetics of drug metabolism enzymes. METHODS: The extent of nonspecific binding to human liver microsomes of the acidic drugs caffeine, naproxen, tolbutamide and phenytoin, and of the basic drugs amiodarone, amitriptyline and nortriptyline was investigated. These drugs were chosen for study on the basis of their lipophilicity, charge, and extent of ionization at pH 7.4. The fraction of drug unbound in the microsomal mixture, fu(mic), was determined by equilibrium dialysis against 0.1 M phosphate buffer, pH 7.4. The data were fitted to a standard saturable binding model defined by the binding affinity KD, and the maximum binding capacity Bmax. The derived binding parameters, KD and Bmax, were used to simulate the effects of saturable nonspecific binding on in vitro enzyme kinetics. RESULTS: The acidic drugs caffeine, tolbutamide and naproxen did not bind appreciably to the microsomal membrane. Phenytoin, a lipophilic weak acid which is mainly unionized at pH 7. 4, was bound to a small extent (fu(mic) = 0.88) and the binding did not depend on drug concentration over the range used. The three weak bases amiodarone, amitriptyline and nortriptyline all bound extensively to the microsomal membrane. The binding was saturable for nortriptyline and amitriptyline. Bmax and KD values for nortriptyline at 1 mg ml-1 microsomal protein were 382 +/- 54 microM and 147 +/- 44 microM, respectively, and for amitriptyline were 375 +/- 23 microM and 178 +/- 33 microM, respectively. Bmax, but not KD, varied approximately proportionately with the microsome concentration. When KD is much less than the Km for a reaction, the apparent Km based on total drug can be corrected by multiplying by fu(mic). When the substrate concentration used in a kinetic study is similar to or greater than the KD (Km >/= KD), simulations predict complex effects on the reaction kinetics. When expressed in terms of total drug concentrations, sigmoidal reaction velocity vs substrate concentration plots and curved Eadie Hofstee plots are predicted. CONCLUSIONS: Nonspecific drug binding in microsomal incubation mixtures can be qualitatively predicted from the physicochemical characteristics of the drug substrate. The binding of lipophilic weak bases is saturable and can be described by a standard binding model. If the substrate concentrations used for in vitro kinetic studies are in the saturable binding range, complex effects are predicted on the reaction kinetics when expressed in terms of total (added) drug concentration. Sigmoidal reaction curves result which are similar to the Hill plots seen with cooperative substrate binding. PMID- 10792205 TI - The risk of acute pancreatitis associated with acid-suppressing drugs. AB - AIMS: To assess the risk of acute pancreatitis associated with use of acid suppressing drugs. METHODS: We conducted a retrospective cohort study with a nested case-control design within the General Practice Research Database (GPRD) in the United Kingdom. The cohort included 180 178 persons aged 20-74 years, who had received at least one prescription of cimetidine, famotidine, nizatidine, ranitidine, lansoprazole, or omeprazole from January 1992 to September 1997 and who did not have major risk factors for pancreatic diseases. Patients with a computerized medical history compatible with idiopathic acute pancreatitis were validated through review of medical records. For the nested case-control analysis 1000 controls were randomly selected from the study population. RESULTS: We identified 88 potential cases of idiopathic acute pancreatitis. Medical records were available for 86. After review of these records 36 cases of acute pancreatitis were confirmed. Seven cases occurred during nonuse, corresponding to a background incidence rate (IR) of 4.4/100 000 person-years (PY). Six cases occurred during current use of ranitidine (IR 10.5/100 000 PY), five patients were current users of cimetidine (IR 13.9/100 000 PY), and three were current users of omeprazole (IR 7.8/100 000 PY). There were no cases among current users of famotidine, lansoprazole, or nizatidine. Relative risk (RR) compared with nonuse and corrected for age, gender, calendar year and use of medication known to be associated with acute pancreatitis was 1.3 (95% CI: 0.4,4.1) for ranitidine, 2.1 (95% CI: 0.6,7.2) for cimetidine, and 1.1 (95% CI: 0.3,4.6) for omeprazole. CONCLUSIONS: The results of this study do not support an association between acute pancreatitis and the use of acid-suppressing drugs, although a substantial increase in risk cannot be excluded with confidence. PMID- 10792206 TI - Graft material and results of platelet inhibitor trials in peripheral arterial reconstructions: reappraisal of results from a meta-analysis. AB - AIMS: To investigate the characteristics of published trials in order to establish the origin of the differing results obtained in trials of platelet inhibitors after peripheral bypass procedures. METHODS: Analysis of the information from 11 randomised, controlled trials of platelet inhibitors after peripheral bypass procedures published up until 1999 and involving 2302 patients undergoing peripheral bypass operations, 1250 of whom were treated with platelet inhibitors. RESULTS: There is a significant treatment benefit of platelet inhibitors on meta-analysis of the trials, but a significant heterogeneity amongst the individual trial results. The proportion of patients in a trial with prosthetic grafts was a significant factor in explaining the heterogeneity. Proportion of prosthetic grafts was associated with sample size and with the proportion of grafts above the knee, but these were not found to make an independent contribution to the heterogeneity observed. The platelet inhibitor regimen used, the severity of ischaemic symptoms and the proportion of smokers included were also not found to be important. CONCLUSIONS: The improvement of graft patency by aspirin and related platelet inhibitors in clinical trials in peripheral bypass procedures can be attributed to an effect on patients with prosthetic grafts. There is little evidence that these agents prevent occlusion of vein grafts. The conclusion of an earlier meta-analysis that antiplatelet agents should be used for all bypasses is not supported. PMID- 10792207 TI - A discordance of the cytochrome P450 2C19 genotype and phenotype in patients with advanced cancer. AB - AIMS: To examine the relationship between cytochrome P450 2C19 (CYP2C19) genotype and expressed metabolic activity in 16 patients with advanced metastatic cancer. METHODS: Individual CYP2C19 genotypes were determined by PCR-based amplification, followed by restriction fragment length analysis, and compared with observed CYP2C19 metabolic activity, as determined using the log hydroxylation index of omeprazole. RESULTS: All 16 patients had an extensive metabolizer genotype. However, based on the antimode in a distribution of log omeprazole hydroxylation indices from healthy volunteers, four of the patients had a poor metabolizer phenotype and there was a general shift of the remaining 12 patients towards a slower metabolic phenotype. This suggests a reduction in metabolic activity for all patients relative to healthy volunteers. A careful analysis of patient medical records failed to reveal any drug interactions or other source for the observed discordance between genotype and phenotype. CONCLUSIONS: There are no previous reports of a 'discordance' between genotype and expressed enzyme activity in cancer patients. Such a decrease in enzyme activity could have an impact on the efficacy and toxicity of chemotherapeutic agents and other drugs, used in standard oncology practice. PMID- 10792208 TI - Is the use of conjugated equine oestrogens in hormone replacement therapy still appropriate? PMID- 10792209 TI - Mohs' micrographic surgery for cutaneous squamous cell carcinoma: practical considerations. PMID- 10792210 TI - Thyroid hormone and hair growth. PMID- 10792211 TI - Stealth tactics of staphylococci. PMID- 10792212 TI - Is morphoea caused by Borrelia burgdorferi? A review. AB - The aetiology of morphoea and lichen sclerosus et atrophicus is still unknown. Since the detection of Borrelia burgdorferi (B. burgdorferi) as the causative agent of Lyme disease, there has been debate about a possible association between B. burgdorferi and morphoea. Initial serological and cultural studies showed controversial results. The introduction of polymerase chain reaction (PCR) initially suggested an association between B. burgdorferi and morphoea. We reviewed the literature on B. burgdorferi (specific serology, immunohistology, culture, lymphocyte stimulation and DNA detection by PCR) since 1983, using Medline and Current Contents. Histological and immunohistological detection of B. burgdorferi was reported in 0-40% (20 of 82) of the cases with morphoea and in 46 50% (17 of 36) of the cases with lichen sclerosus et atrophicus. Cultivation of spirochetes from lesional skin succeeded in five patients (five of 68) with morphoea, but failed in patients with lichen sclerosus et atrophicus. In Europe and Asia, serological detection of antibodies against B. burgdorferi was described in 0-60% (138 of 609) of patients with morphoea and in 19% (six of 32) in the U.S.A. For lichen sclerosus et atrophicus 0-25% of the published cases (three of 23) in Europe and Asia were seropositive. DNA from B. burgdorferi was detected by PCR in 0-100% (17 of 82) of the tissues of patients with morphoea in Europe and Asia, but not a single case among 98 patients was reported to be positive from the U. S.A. In Europe and Asia, borrelial DNA was detected in 0 100% (nine of 28) of the cases with lichen sclerosus et atrophicus, whereas in the U.S.A. none of 48 patients was positive. There are two possible explanations for these contradictory findings: the most likely is that B. burgdorferi is not a causative agent for morphoea. Another possible explanation could be that a subset of morphoea is caused by a special subspecies of B. burgdorferi that is present in Europe and Asia but does not occur in the U.S.A. PMID- 10792213 TI - Thyroid hormone receptor beta1 is expressed in the human hair follicle. AB - To understand better the mechanisms by which thyroid hormone can exert its effects on the hair follicle, we looked for the expression of members of the thyroid hormone receptor (TR) family in human hair follicles. Immunoreactive TRs were detected in both dermal and epithelial compartments of the human pilosebaceous unit. Using reverse transcriptase-polymerase chain reaction, we established that TRbeta1 was the predominant form of TR expressed in the human hair follicle. In addition, we investigated the effects of 3,3', 5-triiodo-L thyronine (T3) on the survival of human hair follicles in vitro, to understand the role of this thyroid hormone on hair follicle homeostasis. A physiological level of free T3 significantly enhanced human hair survival in vitro. PMID- 10792214 TI - Protective role of nitric oxide-mediated inflammatory response against lipid peroxidation in ultraviolet B-irradiated skin. AB - Ultraviolet (UV) irradiation is known to induce serious oxidative damage in the skin via lipid peroxidation. Nitric oxide (NO) synthesized by keratinocytes, melanocytes and endothelial cells in response to proinflammatory cytokines and UV radiation, has been reported to prevent UV-induced apoptosis in the skin. We have examined the effects of NO on UVB-induced lipid peroxidation in murine skin in vivo. UVB induced a dose-dependent increase in lipid peroxidation of skin extracts in vitro; however, lipid peroxidation in the skin in vivo remained unaffected at irradiation doses of less than 1.0 J cm-2 and decreased significantly at doses over 1.5 J cm-2 (P < 0.01). Time-delayed inhibition of lipid peroxidation in the skin in vivo was observed after irradiation at 1.5 J cm 2. Administration of N G-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis, enhanced lipid peroxidation (P < 0.05), while it suppressed the ear swelling response (ESR), a biological marker of inflammation. By contrast, administration of sodium nitroprusside, an NO enhancer, suppressed lipid peroxidation (P < 0. 01), while it enhanced the ESR. Expression of inducible nitric oxide synthase (iNOS) was observed from 12 to 48 h postirradiation at doses of 0.4-1.6 J cm-2. The UVB-induced iNOS expression was markedly inhibited by L-NAME, suggesting that iNOS is a major enzyme in the production of NO. These results suggest that NO acts as a mediator of the inflammatory response in UVB irradiated skin, and that lipid peroxidation is inversely regulated with the NO mediated inflammatory response in vivo. PMID- 10792215 TI - Ultraviolet B radiation exerts enhancing effects on the production of a complement component, C3, by interferon-gamma-stimulated cultured human epidermal keratinocytes, in contrast to photochemotherapy and ultraviolet A radiation that show suppressive effects. AB - The present study was designed to investigate by using enzyme-linked immunosorbent assay and reverse transcriptase-polymerase chain reaction methods whether photochemotherapy (PUVA) or ultraviolet (UV) B treatment affects C3 production by interferon (IFN)-gamma-stimulated keratinocytes cultured in serum free medium. The results showed that PUVA and UVA reduced C3 production by IFN gamma-stimulated epidermal keratinocytes dose-dependently, although the effect of PUVA was stronger than that of UVA alone. Interestingly, UVB induced an enhancement of C3 production at doses ranging from 10 to 50 mJ cm-2. This phenomenon was found at both the protein and mRNA levels. In every experiment, changes in C3 mRNA levels preceded those in its protein levels. Reduced C3 production at higher doses of 75 and 100 mJ cm-2 were probably due to cytotoxic effects of UVB. In our experimental system, PUVA, UVA or UVB treatment did not affect C3 production without IFN-gamma stimulation. Our results suggest that a reduction in C3 production by PUVA treatment may in part explain the efficacy of PUVA in the treatment of inflammatory dermatoses such as psoriasis, while the results of the UVB experiments may partially explain the proinflammatory nature of UVB. PMID- 10792216 TI - Atopic dermatitis-like symptoms in hypomagnesaemic hairless rats are prevented and inhibited by systemic or topical SDZ ASM 981. AB - Magnesium deficiency in hairless rats results in a transient erythematous rash within several days, the pathogenetic mechanisms of which are not yet well defined. However, the extremely pruritic rash closely mimics the acute clinical features of atopic dermatitis. Owing to the similarity of clinical signs between hypomagnesaemic rats and patients with atopic dermatitis, this rodent skin condition holds promise as a model for the in vivo evaluation of new treatment modalities against pruritic inflammatory skin conditions. The efficacy of the new ascomycin macrolactam derivative SDZ ASM 981 was tested in hypomagnesaemic rats by systemic or topical administration using prophylactic or therapeutic treatment regimens. Oral treatment of diseased rats with SDZ ASM 981 (12.5 mg kg-1 daily) inhibited the erythematous pruritic rash within 1 day after the start of treatment. This was associated with a clear reduction in histaminaemia, leucocytosis, eosinophilia and serum nitric oxide levels. The same daily oral dose of SDZ ASM 981 administered before the onset of the rash proved to be an efficacious prophylactic treatment regimen to prevent signs. Topical treatment of the ears with 0.4% SDZ ASM 981 locally inhibited and prevented inflammatory changes in a therapeutic and prophylactic treatment regimen, respectively. The histo- and immunopathological skin changes, as well as the numbers of degranulated mast cells in the dermis, were reversed towards normal after oral and topical administration. The pharmacological activity of SDZ ASM 981 reported here corresponds well to its anti-inflammatory and antipruritic activity observed in atopic dermatitis patients, confirming the usefulness of this rat model in drug evaluations. PMID- 10792217 TI - Effects of two novel cationic staphylococcal proteins (NP-tase and p70)and enterotoxin B on IgE synthesis and interleukin-4 and interferon-gamma production in patients with atopic dermatitis. AB - We have characterized the cell-mediated and humoral immune response of patients with atopic dermatitis (AD) and healthy controls in response to two novel staphylococcal antigens (NP-tase, p70) and the superantigen staphylococcal enterotoxin B (SEB). The parameters studied were IgE, interleukin (IL)-4 and interferon (IFN)-gamma synthesis by peripheral blood mononuclear cells (PBMC) after stimulation with NP-tase, p70 and SEB in vitro. Both antigens, as well as SEB, induced IL-4 and IFN-gamma secretion in patients and controls. However, patients with AD showed a significantly diminished IFN-gamma production in response to NP-tase or SEB. Furthermore, we demonstrated a good correlation between antigen-stimulated IgE production and the IL-4/IFN-gamma ratio in vitro. A distinct subgroup of PBMC showed impaired IFN-gamma synthesis and enhanced IL-4 secretion after incubation with p70 or NP-tase. These data support evidence that a subgroup of patients with AD, synthesizing low levels of IFN-gamma after stimulation with staphylococcal antigens, may have impaired abilities to clear Staphylococcus aureus colonization. Persistent staphylococcal antigens could then be responsible for inflammatory and allergic skin reactions in patients with AD. We therefore conclude that, besides superantigens, staphylococcal antigens may also play a part in the pathogenesis of AD. PMID- 10792218 TI - DNA image cytometry in malignant and benign sweat gland tumours. AB - The histopathological differentiation between well-differentiated carcinomas and atypical adenomas of sweat gland origin may be difficult, even if immunohistochemical methods are used. Therefore, additional techniques may be helpful. We previously demonstrated that DNA image cytometry (ICM-DNA) can be useful in distinguishing between malignant and benign clear cell hidradenoma. In the present study, a larger series of sweat gland tumours, with a clear-cut diagnosis as malignant or benign on histopathological criteria, was examined by ICM-DNA. Enzymatic cell separation specimens were prepared from paraffin-embedded tissues of 18 sweat gland carcinomas (14 porocarcinomas, one classic eccrine adenocarcinoma, two microcystic adnexal carcinomas and one mostly ductal apocrine carcinoma) and 47 benign sweat gland tumours (three syringocystadenomas, five spiradenomas, 14 cylindromas, three syringomas, seven nodular hidradenomas, 10 cutaneous mixed tumours, four poromas and one apocrine hidrocystoma). Specimens were examined by ICM-DNA according to the current recommendations of the European Society for Analytical Cellular Pathology with the AutoCyte QUIC-DNA workstation using mesenchymal cells as an internal reference. DNA aneuploidy was detected by the stemline interpretation according to Bocking and/or at least three 5[c] exceeding events. DNA aneuploidy was detected in 16 of 18 (89%) of the sweat gland carcinomas, but in none of the 47 adenomas. These results suggest that the detection of DNA aneuploidy in sweat gland tumours using ICM-DNA is a clear and specific indicator of prospective malignancy. PMID- 10792219 TI - The temporal and spatial distribution of p21WAF expression in skin appendages. AB - p21WAF is a cyclin-dependent kinase inhibitor which is widely expressed in epidermal structures. Using a combination of double immunocytochemical staining and combined in situ hybridization, we show that there is a striking exclusivity between the expression of Ki67 and p21WAF in the hair matrix. Some cells that are Ki67-positive also express p53, but as they exit the cell cycle they assume p21WAF-positive/p53-negative status. By contrast, cells in the interfollicular epidermis of psoriatic lesions, in the sebaceous gland, and in the outer root sheath are p21WAF-positive/p53-positive but Ki67-negative. These results suggest that in some anatomical parts of the epidermis, p21WAF expression can accompany p53 expression, whereas in other parts, the expression of these markers is reciprocal, suggesting that other pathways may be controlling p21WAF expression. In order to define, functionally, the presence of p53-independent p21WAF expression in skin, we examined lesions of Bowen's disease in which both alleles of p53 were inactivated. p21WAF expression was still observed, confirming a role for p53-independent expression of p21WAF in human skin in vivo. PMID- 10792221 TI - Is there a geographical variation in eczema prevalence in the UK? Evidence from the 1958 British Birth Cohort Study. AB - Some studies have suggested that the prevalence of atopic eczema may vary between geographical regions. This descriptive study investigates the regional and subregional geography of reported and examined eczema prevalence at the age of 7, 11 and 16 years in Britain using data from the 1958 birth cohort study (n = 828). Estimates of the relative risk of reported eczema associated with residence in each region of the country were calculated and the regional distribution of reported and examined eczema prevalence was compared. The reported prevalence of eczema was mapped at the smaller county level. Comparisons were made with the county-level distribution of asthma and hay fever prevalence. The study showed a marked and statistically significant variation in eczema prevalence across the regions in Britain which was present for examined as well as reported eczema. The highest risk was associated with four regions: North Midlands; Eastern; London and the South-East; and Southern. This regional pattern was not altered significantly after adjustment for social class and family size. The geographical distribution of eczema prevalence was largely maintained when analysed at the county level. Few similarities were found between the county-level distribution of eczema prevalence and that for asthma and hay fever. Explanations for this strong regional variation now need to be sought in terms of environmental and life-style associations. PMID- 10792220 TI - Dissociation of intra- and extracellular domains of desmosomal cadherins and E cadherin in Hailey-Hailey disease and Darier's disease. AB - In order to clarify the pathomechanism of acantholysis in Hailey-Hailey disease (HHD) and Darier's disease (DD), the distribution of desmosomal and adherens junction-associated proteins was studied in the skin of patients with HHD (n = 4) and DD (n = 3). Domain-specific antibodies were used to determine the cellular localization of the desmosomal transmembrane glycoproteins (desmogleins 1 and 3 and desmocollin), desmosomal plaque proteins (desmoplakin, plakophilin and plakoglobin) and adherens junction-associated proteins (E-cadherin, alpha catenin, beta-catenin and actin). A significant difference in staining patterns between intra- and extracellular domains of desmosomal cadherins and E-cadherin was demonstrated in acantholytic cells in both HHD and DD, but not in those in pemphigus vulgaris and pemphigus foliaceus samples used as controls. In acantholytic cells in HHD and DD, antibodies against attachment plaque proteins and intracellular epitopes of desmosomal cadherins exhibited diffuse cytoplasmic staining, whereas markedly reduced staining was observed with antibodies against extracellular epitopes of the desmogleins. Similarly, membrane staining of an intracellular epitope of E-cadherin was preserved, while immunoreactivity of an extracellular epitope of E-cadherin was destroyed. While the DD gene has been identified as ATP2A2, the gene for HHD has not been clarified. The dissociation of intra- and extracellular domains of desmosomal cadherin and E-cadherin is characteristic of the acantholytic cells in HHD and DD, and not of pemphigus. This common phenomenon in HHD and DD might be closely related to the pathophysiological mechanisms in both conditions. PMID- 10792222 TI - The British Association of Dermatologists audit of atopic eczema management in secondary care. Phase 3: audit of service outcome. AB - Service outcome was examined by a preconsultation (part 1) and a 6-week postconsultation (part 2) patient questionnaire in 29 hospital dermatology departments randomly selected from an original sample of 187 centres across the U.K. The outcome measures were: quality of life as measured by the Dermatology Life Quality Index (DLQI) and Children's DLQI (CDLQI), improvement in sleep loss, improvement in worse aspect of skin disease and return to work or school. Three hundred and fifty-two questionnaires (115 adults, 237 children) were completed for part 1, and 235 (67%) replied to part 2. The mean DLQI at initial consultation was 12.5, dropping to 9.7 at 6 weeks (P = 0.001). The mean CDLQI at initial consultation was 10.5, dropping to 8.7 at 6 weeks (P < 0.001). Forty-nine per cent of adults and 44% of children had a > 25% relative improvement in score, which did not meet the 60% working standard. Forty-four per cent of adults and 47% of children had an improvement in sleep loss at 6 weeks, falling short of the 70% working standard. Sixty-one per cent of adults and 59% of children had an improvement in the worst aspect of their skin condition at 6 weeks, falling short of the 80% working standard. Of the 20 adults and eight children off work/school during part 1, 70% of adults and 87.5% of children had returned to work/school by 6 weeks. This met the 80% working standard for children but not for adults. On a national scale, only one of the eight working standards for service outcome was met, although most of the working standards were met by at least one of the 11 National Health Service administrative areas. This study presents the first national data on the outcomes of a representative sample of atopic eczema patients seen in secondary care. Small sample sizes, instruments which may be insensitive to change, as well as local factors such as case-mix, baseline severity and staff to patient ratios need to be taken into account when interpreting these results. Nevertheless, the results of this baseline audit suggest that the outcome of patients with atopic eczema following secondary care consultation may not be as good as some doctors believe. This suggests that an improvement in practice, a re-evaluation of the working standards, or both, is needed and should be examined in future audit cycles. PMID- 10792223 TI - The Salford Psoriasis Index: an holistic measure of psoriasis severity. AB - We have developed, tested and validated a new scoring system for psoriasis: the Salford Psoriasis Index (SPI). The SPI incorporates the current clinical extent of psoriasis based on the Psoriasis Area and Severity Index (PASI), a score indicating psychosocial disability, and past severity based on treatment history. The resultant three-figure SPI (signs, psychosocial disability, interventions) is a similar paradigm to the TNM (tumour, nodes, metastasis) classification used for cancer staging. The first figure transforms the PASI into a number from 0 to 10 reflecting extent of psoriasis. The second assesses the psychosocial impact of psoriasis on each patient using a 0-10 visual analogue scale. The third figure reflects historical severity of disease as judged by the need for systemic treatment, admission to hospital and number of episodes of erythroderma. The SPI was prospectively employed in assessing 150 consecutive patients with psoriasis. Furthermore, in a separate cohort of 100 patients we tested the Psychosocial Impact Score against a recognized self-report psoriasis-specific measure, the Psoriasis Disability Index. There was a strong correlation between the two (r = 0.59, P < 0.001). However, the Psychosocial Impact Score correlated poorly with clinical extent scores such as the PASI (r = 0.28, P < 0.05) and the Self administered PASI in 72 patients tested (r = 0.19, P = 0.1). There was a high correlation between all six observers in 20 patients for both PASI (r = 0.71; 95% confidence interval, CI 0.51-0.86) and the Extent Score (r = 0.70; 95% CI 0. 56 0.89). We believe that the SPI will be more relevant to real-life categorization of psoriasis severity in that it takes an holistic approach based not only on physician assessment but also psychological disability and treatment resistance. PMID- 10792224 TI - Psoralen-ultraviolet A-induced erythema: sensitivity correlates with the concentrations of psoralen in suction blister fluid. AB - Since the advent of psoralen-ultraviolet A (PUVA) therapy, the value of plasma 8 methoxypsoralen (8-MOP) concentrations to predict PUVA-induced erythema has been widely investigated. Plasma 8-MOP concentrations have not been proportional to, and cannot alone predict, the degree of PUVA-induced erythema. We assumed that PUVA-induced erythema was related more closely to psoralen concentrations in the skin tissue rather than those within blood vessels. This study was designed to investigate the correlations between the 8-MOP concentrations in suction blister fluid (SBF) and in plasma, with the degree of PUVA-induced erythema. 8-MOP concentrations in plasma and SBF were measured in 15 vitiligo patients and 11 volunteers. Blood and SBF samples were collected 2 h after taking 8-MOP, and 8 MOP concentrations in plasma and SBF were quantified using reverse-phase high performance liquid chromatography. Eleven volunteers were phototested using a series of doses of ultraviolet A at the time of sampling. The erythema responses were estimated visually to determine the minimal phototoxic dose (MPD). SBF 8-MOP concentrations showed a weak positive correlation with plasma 8-MOP concentrations, which means that we could not predict the exact SBF 8-MOP concentrations using the plasma 8-MOP concentrations. The MPD showed a better correlation with the log of the SBF 8-MOP concentration than with that of the plasma 8-MOP concentration. These results show that plasma 8-MOP concentration cannot represent the exact SBF 8-MOP concentration, and that SBF 8-MOP concentrations, which are representative of the skin tissue 8-MOP level, are more closely related to the erythemal sensitivity during PUVA therapy. PMID- 10792225 TI - Evaluation of a multicentre study of synchronous application of narrowband ultraviolet B phototherapy (TL-01) and bathing in Dead Sea salt solution for psoriasis vulgaris. AB - The synchronous application of narrowband UVB phototherapy with 311 nm lamps (Philips TL-01) and bathing in Dead Sea salt solution was evaluated in a multicentre trial (n = 60) in outpatients suffering from psoriasis vulgaris. The study design consisted of an initial therapy phase of up to 35 treatments (three to five times a week) followed by maintenance therapy with up to 35 further applications (once or twice a week). Evaluation was performed separately for patients in according-to-protocol (ATP) (n = 280) and intention-to-treat (ITT) (n = 692) groups. An overall significant improvement of the Psoriasis Area and Severity Index (PASI) score (P < 0.05) could be shown for both groups during initial therapy with 71.4% improvement for ATP and 61% for ITT patients. The mean PASI for ATP (values for ITT in parentheses) was 17.7 (18.6) at baseline, 9.5 (10.7) after 20 applications and 5.2 (7.4) at the end of initial therapy. On average, ATP patients received 3.9 (3.5) applications per week with a cumulative irradiation dose of 19.5 J cm-2 (16.2 J cm-2). The most frequent side-effect was erythema, observed in 8.7% of the patients. Subjective evaluation of the therapy by the patients (n = 168) was excellent. Seventy-nine per cent of patients preferred the new treatment strategy in comparison with other previous therapies and 88% regarded this therapy as pleasant and comfortable. In conclusion, we could demonstrate a significant effect of therapy in both the ATP and the ITT groups for this new treatment system which imitates, as far as possible, the Dead Sea climatic conditions, with no severe side-effects and a high acceptance by the patients. PMID- 10792226 TI - Measurement of patient dose in ultraviolet therapy using a phantom. AB - A phantom was developed as a reproducible means of measuring the irradiance in an occupied ultraviolet cabin, by placing the phantom, or replica person, in the cabin, obviating the need for human exposure. The contributions to the patient irradiance measured in the cabin were investigated, looking in particular at the effect of the reflectors. Radiation undergoing single reflection was seen to contribute to a greater extent than multiple reflections. Placing an object in the cabin reduces the measured irradiance due to the blocking of multiple reflections, but variation in the exact shape and size of the object has less effect, which is useful as patients are of all shapes and sizes and a representative phantom was to be developed. The phantom was made of expanded polystyrene blocks with an embedded probe. Measurements were made to verify the equivalence of human and phantom cabin occupancy. It was found that the irradiance measured with the phantom in the cabin lies within the values measured with human occupancy. PMID- 10792227 TI - A retrospective study of outcome of Mohs' micrographic surgery for cutaneous squamous cell carcinoma using formalin fixed sections. AB - The surgical management of recurrent or large squamous cell carcinoma (SCC) can be challenging as tumours often extend beyond visible margins. Micrographic surgery is a potentially effective method of ensuring complete clearance of tumour. A retrospective study of all cases of SCC treated by micrographic surgery in this department between 1986 and 1996 has been done. Sixty-one patients were treated using a formalin-fixed paraffin-embedded tissue technique with a median follow-up of 4 years. In two cases there was local recurrence and in three others metastasis to local lymph nodes. The overall cure rate was 92% (56 of 61), which compares favourably with published series using chemosurgery and frozen tissue techniques. The results show that this technique of micrographic surgery is a satisfactory and cost-effective alternative to conventional frozen section techniques in the treatment of SCC. The formalin-fixed tissue method has the advantage of providing high-quality permanent histological sections using existing conventional pathology services. PMID- 10792228 TI - In vitro susceptibility of the seven Malassezia species to ketoconazole, voriconazole, itraconazole and terbinafine. AB - Fifty-five strains, either authentic or ex-type, of seven Malassezia species were investigated for in vitro susceptibility to various concentrations (0.03-64.0 microg/mL) of three azole drugs, ketoconazole, voriconazole and itraconazole, as well as the allylamine terbinafine, using the agar dilution method. All strains of the seven Malassezia species were susceptible to the three azole drugs at low concentrations. M. furfur, M. sympodialis, M. slooffiae, M. pachydermatis, M. globosa, M. obtusa and M. restricta were most sensitive to ketoconazole and itraconazole, with minimum inhibitory concentrations (MICs) ranging from < or = 0.03 to 0.125 microg/mL. The recently introduced antifungal, voriconazole, was also very effective, with MIC80 values < or = 0.03 microg/mL for 80% of strains. MICs of terbinafine against the seven Malassezia species ranged from 30 x 109/l (P < 0.001) in both studies, and by a haemorrhagic score of 3 in study A (P < 0.001). Although the reduction of early fatal haemorrhages was not significant, a substantial clinical improvement was evident in terms of reduction of the severity of bleeding symptoms, blood product consumption and overall induction mortality when ATRA was combined with idarubicin. PMID- 10792271 TI - Histone deacetylase inhibitor but not arsenic trioxide differentiates acute promyelocytic leukaemia cells with t(11;17) in combination with all-trans retinoic acid. AB - Acute promyelocytic leukaemia (APL) with t(11;17)/PLZF-RARalpha responds poorly to all-trans retinoic acid (ATRA) and arsenic trioxide (As2O3), in contrast to APL with t(15;17)/PML-RARalpha. Molecular studies have shown that histone deacetylase (HDAC) recruited by PLZF-RARalpha is associated with the ATRA resistance. Here, we analysed in vitro the differentiation of APL cells with t(11;17) using ATRA, As203, granulocyte colony-stimulating factor (G-CSF), HDAC inhibitor trichostatin A (TSA), or combinations of these. Although 1 microM ATRA, which stimulated the differentiation of APL cells with t(15;17), was insufficient to induce differentiation, 3 microM ATRA induced terminal differentiation into granulocytes. As203 alone or in combination with ATRA induced neither differentiation nor apoptosis. However, the combination of TSA and 1 microM ATRA had a potent differentiating effect, although TSA alone had little effect. The combination of 1 microM ATRA and G-CSF did not induce differentiation. These results indicate that APL cells with t(11;17) need a higher concentration of ATRA than those with t(15;17) to differentiate and suggest that HDAC inhibitor is a promising differentiation enhancer in APL with t(11;17). PMID- 10792272 TI - P-glycoprotein (PGP), lung resistance-related protein (LRP) and multidrug resistance-associated protein (MRP) expression in acute promyelocytic leukaemia. AB - We analysed the expression of three drug transporter proteins [p-glycoprotein (PGP), lung resistance-related protein (LRP) and multidrug resistance-associated protein (MRP1)] involved in anthracycline resistance that are frequently overexpressed in poor-risk adult acute non-lymphocytic leukaemia (ANLL), in 23 acute promyelocytic leukaemia (APL) patients at onset managed at a single institution. Cellular daunorubicin accumulation was also evaluated. At onset, no case had PGP or MRP1 expression that exceeded that of non-multidrug-resistant (MDR) cell lines. Only one case showed LRP overexpression. No peculiar MDR features distinguished the seven patients who relapsed from those who maintained complete remission. In the onset vs. first relapse, only one patient showed an increased (threefold) PGP expression at relapse. At second relapse, three out of four patients showed a PGP expression two- to threefold higher than baseline values. These results are consistent with the view that low PGP, LRP and MRP1 expression and the absence of defects in intracellular drug accumulation may account for the peculiarly high sensitivity of APLs to anthracycline. It does not support the screening of MDR markers in APL patients at onset as predicting factors of early relapse. The results suggest that no significant changes in PGP, LRP or MRP1 expression are likely to occur at first relapse. In contrast, PGP expression is likely to increase later in the patient history as a result of additional chemotherapy courses. PMID- 10792273 TI - Flow cytometric assessment of CD15+CD117+ cells for the detection of minimal residual disease in adult acute myeloid leukaemia. AB - There is little information available regarding immunophenotypic monitoring of minimal residual disease (MRD) in acute myeloid leukaemia (AML). We investigated leukaemic cells co-expressing CD15 and CD117 (CD15+CD117+) in 72 adult AML cases at diagnosis. In 22 cases (31%) with various AML subtypes, more than 5% of leukaemic cells showed the CD15+CD117+ phenotype (range 5.22-55.48%). These 22 cases were younger and had a higher complete remission (CR) rate than the other AML cases, but the CD15+CD117+ cell percentage at diagnosis showed no correlation with the CR duration among the 72 cases. The CD15+CD117+ cell percentage showed a range of 0.00-0.08% in bone marrow cells from 10 haematologically normal subjects. We also investigated CD15+CD117+ cells in sequential bone marrow samples from 17 AML patients who achieved CR and who had had more than 5% CD15+CD117+ leukaemic cells at diagnosis. Because the CD15+CD117+ cell percentage varied among these AML cases, we calculated the percentage of MRD ?MRD% = [CD15+CD117+ cells (%) in each sequential marrow sample] / [CD15+CD117+ cells (%) at diagnosis of the corresponding case] x 100?. A high MRD% after 10 months of CR was significantly associated with a short CR duration (P = 0.0004), whereas continuation of a well-reduced MRD% was associated with a long CR duration. The leukaemic cells conserved the CD15+CD117+ phenotype in all of the eight cases who relapsed. Flow cytometric monitoring of CD15+CD117+ cells is simple and can be applied to a substantial fraction of AML cases. This monitoring may be useful for predicting relapse of adult AML. PMID- 10792274 TI - Clinicopathological features of aggressive large granular lymphocyte leukaemia resemble Fas ligand transgenic mice. AB - Fas ligand triggers cell death after interaction with its receptor Fas. Altered expression of Fas has been associated with lymphoproliferation and autoimmune disorders in both mice and man. Apoptosis of lung and liver tissue is seen in Fas ligand transgenic mice. It is not known whether constitutive expression of Fas ligand can cause a similar human disease. Four patients with aggressive large granular lymphocyte (LGL) leukaemia involving lung and liver were studied. All four patients were severely ill with pulmonary involvement. Two patients presented with hypoxia and were oxygen dependent; the other two patients had severe pulmonary hypertension. Lung biopsies showed interstitial infiltration by leukaemic LGL. The infiltrating lymphocytes expressed both Fas and Fas ligand, whereas normal pneumocytes expressed only Fas. Similar findings were observed in liver biopsies from these patients. Features mimicking the pathological changes of graft-versus-host disease were observed, including pneumocyte apoptosis. All four patients had high levels of circulating Fas ligand. Successful treatment with oral methotrexate or 2-chlorodeoxyadenosine was associated with disappearance or marked reduction of circulating Fas ligand. These results suggest that dysregulated expression of Fas ligand can lead to human disease with pathological features resembling graft-versus-host disease. PMID- 10792275 TI - Two types of acquired idiopathic sideroblastic anaemia (AISA): a time-tested distinction. AB - In 1982, acquired idiopathic sideroblastic anaemia (AISA) was included by the French-American-British (FAB) Co-operative Group in their classification of myelodysplastic syndromes (MDS). However, the malignant potentiality of AISA has always been a matter of debate. In different series, median survival and rates of transformation into acute myeloid leukaemia (AML) varied extensively. On cytomorphological grounds, AISA can be divided into pure (dyserythropoietic) sideroblastic anaemia (PSA), in which dysplasia is confined to erythropoietic cells, and a true myelodysplastic form (RARS), which is characterized by additional dysplastic features of granulopoiesis and/or megakaryopoiesis. In a previous study, based on retrospective analysis of 94 patients with AISA, we found that both types of sideroblastic anaemia differed considerably in terms of survival and risk of AML transformation. Almost identical results have now been obtained through a prospective study of 232 new patients with AISA. The difference in survival between PSA and RARS remained significant over the whole period of follow-up (survival after 3 years being 77% vs. 56%; P = 0.003), and the incidence of AML did not increase with time in the PSA group, even in the long term. This prospective study strongly supported our conclusion that cytomorphological distinction between PSA and RARS provides valuable prognostic information. PMID- 10792276 TI - Mastocytosis cells bearing a c-kit activating point mutation are characterized by hypersensitivity to stem cell factor and increased apoptosis. AB - Mastocytosis is characterized by abnormal infiltration of mast cells into various organs. An activating mutation in c-kit, involving an A --> T substitution at nucleotide 2648 has recently been described in some patients with mastocytosis. We describe a 12-year-old girl with this mutation in her bone marrow cells at diagnosis with a myelodysplastic syndrome (MDS) without evidence of mastocytosis, and then in peripheral blood mononuclear cells 1 year later after the emergence of mastocytosis. The role of the c-Kit receptor and its ligand stem cell factor (SCF) in the pathogenesis of the disease was analysed in marrow cell clonogenic assays. We show that the genetic abnormalities in the patient resulted in factor independent growth and hypersensitivity of primitive progenitors to SCF, with increased production of mast cells. Increased apoptosis and cluster formation, consistent with the myelodysplastic nature of the disorder, accompanied accumulation of abnormal cells with increasing concentrations of SCF. PMID- 10792277 TI - Waldenstrom's macroglobulinaemia: a prospective study comparing daily with intermittent oral chlorambucil. AB - This prospective study compared continuous and intermittent chlorambucil therapy of untreated Waldenstrom's macroglobulinaemia. The diagnosis was established by the presence of an IgM monoclonal (M-) protein in the serum, an infiltrate of lymphocytes and plasma cells in the bone marrow, anaemia or other laboratory abnormalities, physical findings or constitutional symptoms. Patients were randomized to receive chlorambucil 0.1 mg/kg/d or chlorambucil 0.3 mg/kg/d orally for 7 d, repeated every 6 weeks. Criteria for response included 50% or more reduction of serum M-protein, increase in haemoglobin level of 2 g/dl without transfusion, >/= 50% decrease of urine M-protein, or a reduction of 2 cm in the size of the liver, spleen or lymph nodes. Forty-six patients were randomized to continuous chlorambucil (n = 24) or to intermittent chlorambucil (n = 22). Nineteen (79%) (95% CI = 58-93) of the 24 patients given continuous therapy had an objective improvement by either reduction of serum M-protein or increase in haemoglobin. Fifteen (68%) (95% CI = 45-86) of the 22 patients given chlorambucil intermittently had an objective response. The size of the liver decreased by >/= 2 cm in 55% of patients, and the size of the spleen decreased >/= 2 cm in 67%. Lymphadenopathy decreased in 71%. Acute leukaemia or refractory anaemia developed in four patients. The median duration of survival was 5.4 years, and there was no difference between the regimens. Chlorambucil is effective for the treatment of Waldenstrom's macroglobulinaemia. Patients must be treated for at least 6 months before therapy is abandoned because response is slow. Chlorambucil is an effective agent and should be compared with purine analogues or rituxan (Rituximab) in a prospective study. PMID- 10792278 TI - Megakaryopoiesis in vitro in myelodysplastic syndromes and acute myeloid leukaemia: effect of pegylated recombinant human megakaryocyte growth and development factor in combination with other growth factors. AB - Pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF) can stimulate megakaryopoiesis in vitro in some myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML) patients. We assessed PEG-rHuMGDF combined with granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage CSF (GM-CSF), interleukin 3 (IL-3), IL6, stem cell factor (SCF) or erythropoietin in 40 MDS, 33 AML and 16 normal bone marrow samples. CD61-positive cells in suspension cultures increased with PEG-rHuMGDF alone in 20/25 RA + RAS, 11/14 RAEB + RAEBt and 29/33 AML cases. Further increases when IL-3 and/or SCF were added to PEG-rHuMGDF occurred in 14/20 RA + RAS, 8/13 RAEB + RAEBt and 18/26 AML cases. CFU-Mk growth was poor overall, but could be enhanced by PEG-rHuMGDF combinations in some patients. Stimulation of megakaryopoiesis by PEG-rHuMGDF can be augmented by IL-3 and SCF in many MDS and AML patients. PMID- 10792279 TI - The effect of granulocyte colony-stimulating factor administration in healthy donors before bone marrow harvesting. AB - To investigate whether granulocyte colony-stimulating factor (G-CSF) administration to donors before harvest may lighten the burden imposed on them and accelerate the bone marrow (BM) recovery, we administered 2 microgram/kg/d of G-CSF for five consecutive days before the marrow harvest. All of the donors tolerated the G-CSF administration well without severe adverse events. After 5 d of G-CSF treatment, CD34+ cells and granulocyte-macrophage colony-forming units (GM-CFU) in the donors' BM exceeded baseline values by 4.2-fold (range 0.71-316) and 1.6-fold (0.28-118) respectively. The concentration of total nucleated cells (x 107/ml) in the graft increased from 1.61 (0.95-3.23) to 2.44 (1.27-4.01). Although we collected 1020 ml of BM and obtained 1.50 x 1010 nucleated cells from unprimed donors, 940 ml of BM were sufficient to obtain 2.14 x 1010 nucleated cells from primed donors. However, G-CSF-primed BM did not shorten the time to tri-lineage engraftment and the duration of hospitalization compared with unprimed BM, although primed BM contained more CD34+ cells than baseline values. We consider that the advantages of BM priming are not the acceleration of BM recovery but rather the reduction of blood loss during BM harvesting. PMID- 10792280 TI - Preliminary experience of allogeneic stem cell transplantation for lymphoproliferative disorders using BEAM-CAMPATH conditioning: an effective regimen with low procedure-related toxicity. AB - Autologous transplantation has an established role in the treatment of lymphoproliferative disorders, but allogeneic transplantation remains controversial. In an attempt to reduce the high procedure-related mortality reported with allografting in lymphoma, we have used BEAM (BCNU, etoposide, cytarabine and melphalan), a standard conditioning regimen for autologous transplantation. As BEAM may be insufficiently immunosuppressive to permit durable engraftment in the allogeneic setting, patients received additional pretransplant immunosuppression with the anti-CD52 antibody CAMPATH-1G from day 5 to day -1. Twelve patients (median age 46 years) underwent allogeneic transplantation for lymphoma (n = 11) or chronic lymphocytic leukaemia (n = 1) from HLA-identical (n = 9) or mismatched (n = 3) sibling donors. Cyclosporin A and methotrexate were used as graft-versus-host disease (GVHD) prophylaxis. One patient died of progressive lymphoma at day +12, the remaining 11 patients engrafted rapidly, with eight demonstrating full donor chimerism. One patient had an episode of rejection and received a further stem cell infusion with sustained recovery. Only one patient developed GVHD (grade I). The low incidence of acute GVHD may be in part related to persisting levels of in vivo CAMPATH-IG at the time of transplantation. Of 11 evaluable patients, nine achieved complete remission (CR), and a further patient achieved CR after donor lymphocyte infusion at 5 months. Our preliminary experience is that this regimen was well tolerated with a low risk of GVHD and appears no more toxic than a BEAM autograft. Further follow-up is required to see whether the low incidence of GVHD impacts upon relapse risk. PMID- 10792281 TI - Characterization of lineage-specific chimaerism in patients with acute leukaemia and myelodysplastic syndrome after allogeneic stem cell transplantation before and after relapse. AB - Recently, we have shown that patients with acute leukaemias and myelodysplastic syndromes (MDS), who showed increasing mixed chimaerism (MC) upon serial PCR analysis after transplant, have a significantly increased risk of relapse. To determine whether the increasing MC in these patients is caused by the reappearance of normal recipient haematopoiesis or by the reoccurrence of malignant cells, we purified different leucocyte subpopulations and analysed these subfractions with regard to their donor-recipient ratio by a PCR-based method for the analysis of minisatellite DNA regions. In 14 patients [eight acute lymphoblastic leukaemia (ALL), three acute myelogenous leukaemia (AML) and three MDS] subfractions were analysed when increasing MC was first noted upon serial analysis of the peripheral blood. In seven of these 14 patients (four ALL, two AML and one MDS), subfractions were characterized at the time of frank haematological relapse. In all 14 patients investigated with increasing MC, recipient cells were detected in different mononuclear cell subpopulations. In patients characterized during frank relapse, two distinct distribution patterns were found. Patients who relapsed before day +300 (one ALL, two AML and one MDS) showed recipient-derived (normal) cells in addition to blast populations in different mononuclear subsets as well as granulocytes. In patients with acute leukaemias who relapsed after day +300 (two ALL and one AML), only leukaemic cells were found that were of recipient origin, whereas all other haematopoietic cell lines were donor derived. These data show that persistent MC in the early post-transplant period is caused predominantly by normal recipient haematopoietic cells. This finding further supports the hypothesis that a state of mixed haematopoietic chimaerism may reduce the clinical graft-versus-leukaemia (GVL) effect of alloreactive donor-derived effector cells in patients with acute leukaemias and MDS, and thus facilitate the proliferation of residual malignant cells that may have survived the preparative regimen. PMID- 10792282 TI - High-dose busulphan alone as cytoreduction before allogeneic or autologous stem cell transplantation for chronic myeloid leukaemia: a single-centre experience. AB - Transplant-related morbidity and mortality remain a major problem following stem cell transplantation (SCT). The use of high-dose single-agent busulphan before allogeneic or autologous SCT may be a reasonable compromise between maintaining cytoreductive efficacy and minimizing toxicity in patients with chronic myeloid leukaemia (CML). Seventy patients with CML have received busulphan as the only cytoreductive therapy before SCT on 79 occasions. The probability of survival of the 14 allogeneic recipients (all of whom were undergoing a second transplant from the original donor) was 78% at 5 years. Sixty-five autologous SCT were performed in 56 patients, of whom 40 were in late chronic phase. The actuarial 3 year post-autograft survival was 54% for these 56 first autografts. For patients in chronic phase, the 3-year survival was 76% compared with 30% at 2 years for those with advanced phase disease. Busulphan therapy was well tolerated, and except for mild mucositis little toxicity was experienced. None of these patients developed hepatic veno-occlusive disease (VOD). Nine patients (one allogeneic and eight autologous recipients) received several doses of an intravenous formulation of busulphan with very low toxicity. High-dose busulphan alone appears sufficient to provide adequate cytoreduction and immunosuppression in second allogeneic transplants and is also effective as cytoreduction before autologous SCT in patients with CML. PMID- 10792283 TI - Expression of HLA-C-specific natural killer cell receptors (CD158a and CD158b) on peripheral blood mononuclear cells after allogeneic bone marrow transplantation. AB - We investigated the expression of natural killer cell receptors (NKRs) for HLA-C on peripheral blood mononuclear cells (PBMCs) in 23 allogeneic bone marrow transplantation (allo-BMT) patients to analyse the role of NKRs in alloresponse concerning graft-versus-host disease (GVHD). CD158a expression was low and there was little change in the expression after allo-BMT. Also, there was no difference in the proportion of CD158a+/CD3- after allo-BMT. In contrast, the proportion of CD158b+/CD3- cells, mainly NK cells, increased in the early stage (< 2 months) after allo-BMT and then gradually decreased (3.3 +/- 2.6% before BMT vs. 15.4 +/- 8. 6% in the early stage after BMT, 8.5 +/- 4.9% during the period 3-6 months after BMT and 7.0 +/- 3.0% > 6 months after BMT; P < 0.05). However, CD158b expression on CD3+ T cells increased 3 months after allo-BMT (1.1 +/- 1.1% before BMT vs. 5.1 +/- 7.7% during the period 3-6 months after BMT and 3.0 +/- 2.4% > 6 months after BMT, P < 0. 05). The highest percentages of CD158 expression in patients without chronic GVHD (cGVHD) and those with cGVHD were compared. The percentage of CD158b+/CD3+ cells and also that of CD158b+/CD8+ cells were significantly increased in patients with cGVHD compared with those in patients without cGVHD (2.6 +/- 2.0% vs. 8.0 +/- 11.2% and 2.3 +/- 1.5% vs. 8.3 +/- 11.7% respectively; P < 0.05). The exact clinical relevance of these CD158b-expressing cells is not clear. However, there is an interesting possibility that CD158b expressing cells play some role in the regulation of GVHD after allo-BMT. PMID- 10792285 TI - Flow cytometric and functional characterization of AC133+ cells from human umbilical cord blood. AB - AC133+ cells may represent an alternative source of transplantable haemopoietic progenitor cells to CD34+ cells. Here, we have addressed the characterization of umbilical cord blood (UCB) AC133+ cells and compared their immunophenotypic and functional features with those of UCB CD34+ cells. UCB AC133+ and CD34+ cell fractions were purified by magnetic cell sorting, analysed by flow cytometry, tested for their content in blast cell colony-forming units (CFU-Bl), erythroid and granulocyte-macrophage colony-forming units before and after expansion in the presence of various haemopoietic growth factor combinations. Median AC133+ cell yield was 62.3%, and median AC133+ population purity was 97.9%. AC133+ cells were found to contain significantly more CFU-Bl than CD34+ cells; furthermore, the replating efficiency, i.e. the number of CFU-Bl capable of generating secondary colonies, was higher in the former than in the latter cells. Both AC133+ and CD34+ cells displayed an increased ability to give rise to committed progenitors after 7-day expansion in liquid cultures. These data suggest that the AC133+ cell subset is a heterogeneous pool of immature and more differentiated cells that can be maintained and expanded in well-defined culture conditions. In comparison with CD34+ cells, UCB AC133+ cells appear to contain a higher number of early haemopoietic progenitors. PMID- 10792284 TI - Reduction of intra- and interlaboratory variation in CD34+ stem cell enumeration using stable test material, standard protocols and targeted training. DK34 Task Force of the European Working Group of Clinical Cell Analysis (EWGCCA). AB - The European Working Group on Clinical Cell Analysis (EWGCCA) has, in preparation for a multicentre peripheral blood stem cell clinical trial, developed a single platform flow cytometric protocol for the enumeration of CD34+ stem cells. Using this protocol, stabilized blood and targeted training, the EWGCCA have attempted to standardize CD34+ stem cell enumeration across 24 clinical sites. Results were directly compared with participants in the UK National External Quality Assessment Scheme (NEQAS) for CD34+ Stem Cell Quantification that analysed the same specimens using non-standardized methods. Two bead-counting systems, Flow Count and TruCount, were also evaluated by the EWGCCA participants during trials 2 and 3. Using Flow-Count, the intralaboratory coefficient of variation (CV) was T) at nucleotide 684 in the gamma common gene. This novel case highlights the need for a thorough evaluation of immunological phenotype and genotype in patients with HLH. PMID- 10792292 TI - Extraordinary bone involvement in a gaucher disease type I patient. AB - We report on a 63-year-old patient with Gaucher disease type I who developed severe bone involvement with destructive lesions and huge soft tissue extension in both humeri that appeared to evolve slowly. The clinical course and histopathological findings in our patient suggested a progressive extraosseous extension of the storage cells into the soft tissue, accompanied by a striking increase of fibrotic tissue and resulting in an impressive deformity. The extraordinary bone involvement in this patient expands our knowledge on the most severe skeletal complications of untreated Gaucher disease. PMID- 10792293 TI - Proliferative involvement of ENX-1, a putative human polycomb group gene, in haematopoietic cells. AB - Homeobox genes have important roles in haematopoiesis and are regulated in an activated state by the trithorax group (trxG) of genes. In a repressed state, they are regulated by the Polycomb group (PcG) of genes. ENX-1, a putative human PcG gene product, interacts with the proto-oncogene product Vav. We report an investigation of the role of ENX-1 in human haematopoiesis. CD34+ cells mobilized to peripheral blood strongly expressed ENX-1. When stimulated to proliferate, both T and B lymphocytes rapidly up-regulated ENX-1. ENX-1 was expressed in all cell lines of the various lineages examined. When HL-60 cells were differentiated to mature granulocytes with all-trans retinoic acid, ENX-1 was down-regulated. Moreover, ENX-1 antisense oligodeoxynucleotide suppressed DNA synthesis in HL-60 cells. Our data indicate that ENX-1 is involved in the proliferation of both normal and malignant haematopoietic cells. PMID- 10792294 TI - Granulocyte colony-stimulating factor (G-CSF) induces the production of cytokines in vivo. AB - Granulocyte colony-stimulating factor (G-CSF) is a haematopoietic growth factor required for the proliferation and differentiation of haematopoietic precursors of neutrophil granulocytes and is now used to overcome congenital and acquired neutropenia. In addition to increasing the numbers of neutrophils in vivo and modulating neutrophil functions, G-CSF may induce the production of cytokines such as tumour necrosis factor alpha (TNF-alpha). In the present study, the plasma levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) in six healthy volunteers given G-CSF at 10 microgram/kg once daily for 6 d were measured and found to be elevated. The elevated levels (P < 0.05) were detected on day 2, peaked on days 6-7 and returned to baseline on day 12. In vitro, G-CSF did not enhance the secretion of TNF-alpha and GM-CSF from mononuclear cells, whole blood or endothelial cells. However, in the co-presence of whole blood and endothelial cells, the secretion of TNF-alpha was significantly enhanced by G-CSF at low concentrations. The GM-CSF secretion, however, was unaltered. G-CSF pretreatment of whole blood suppressed lipopolysaccharide (LPS)-induced secretion of TNF-alpha and GM-CSF in a dose-dependent manner. These results together with our previous findings suggest that G-CSF induces the production of TNF-alpha and GM-CSF in vivo, and that this production may be due to the co-effects of endothelial cells and whole blood under the influence of G-CSF through an as yet unknown network of cells and cytokines. Treatment of whole blood with G-CSF suppresses LPS-induced secretion of TNF-alpha and GM-CSF. PMID- 10792295 TI - Mutation analysis in the HFE gene in patients with hereditary haemochromatosis in Saguenay-Lac-Saint-Jean (Quebec, Canada). AB - A mutation analysis of the HFE gene followed, when applicable, by sequencing was performed on 47 patients with hereditary haemochromatosis (HH) living in Saguenay Lac-Saint-Jean. The C282Y and H63D mutations were present on 50% and 20.3% of the HH chromosomes respectively. These frequencies were very different from those found in other populations and could be, at least partially, the result of a founder effect. No new mutation was identified among the remaining 28.1% of the HH chromosomes. Five of the eight probands with no mutation in the HFE gene had a severe and early onset suggestive of juvenile haemochromatosis. PMID- 10792296 TI - In vivo biotinylation studies: specificity of labelling of reticulated platelets by thiazole orange and mepacrine. AB - Animal in vivo biotinylation studies have demonstrated that thiazole orange (TO) labels the youngest cells in the circulation. TO has since been widely used for the measurement of reticulated platelets. As recent findings suggest that at high concentrations TO also labels platelet dense granules non-specifically, the value of previous work is unclear. Mepacrine also labels platelet dense granules and can detect storage pool defects. In this study, a mouse in vivo biotinylation model was used to determine the specificity of TO and mepacrine staining on platelets recently released into the circulation. The mean life span of biotin/TO (low), biotin/TO (high) and mepacrine/TO dual-positive platelets was 1.4 d (SD 0.5), 2.2 d (SD 0.2) and 2.3 d (SD 0.3) respectively (n = 6) compared with a life span for biotin-positive platelets of 4.9 d (SD 1.6). TO (low), TO (high) and mepacrine labelled 8.0% (SD 3.1), 43.9% (SD 8.3) and 40.0% (SD 9.9) of the total platelet population respectively (results of 30 samples from six mice), which decreased to 6.8% (SD 3. 9), 26.6% (SD 6.9) and 25.7% (SD 10.6) after thrombin degranulation. The shorter life span and lack of thrombin sensitivity of TO (low) positive platelets, suggests that TO (low) measures reticulated platelets specifically. The comparative life spans and thrombin sensitivity of TO (high) and mepacrine-positive platelets suggest that TO (high) labels platelet dense granules. These data also suggest that dense granules are lost during platelet ageing. PMID- 10792297 TI - The G1691 --> A mutation of factor V, but not the G20210 --> A mutation of factor II or the C677 --> T mutation of methylenetetrahydrofolate reductase genes, is associated with venous thrombosis in patients with lupus anticoagulants. AB - Arterial and venous thrombosis are the most common manifestations of antiphospholipid syndrome. To investigate whether genetic determinants contribute to their thrombotic risk, we studied the prevalence of the G1691 --> A mutation in the gene coding for factor V, the G20210 --> A mutation in the prothrombin gene and the C677 --> T mutation in the methylenetetrahydrofolate reductase gene in 152 patients with lupus anticoagulants. One hundred and twenty-eight cases (84%) also had increased titres of anticardiolipin antibodies. History of thrombosis was present in 96 patients (63%); 67 suffered from venous thrombosis only, 23 cases had arterial thrombosis only, six patients had both venous and arterial thrombosis. Five patients were heterozygous for the G1691 --> A mutation in the factor V gene (3%). All of them (100%) suffered from venous thrombosis compared with 68 out of the 147 cases without the mutation (46%) (P = 0.0474). The prevalence of the G20210 --> A mutation in the prothrombin gene was evaluated in 145 patients; eight of these patients were heterozygous (5%). Four of these patients (50%) experienced venous thrombosis compared with 65 out of the 137 patients without the mutation (47%) (P = ns). Neither mutation was associated with arterial thrombotic events. No patient carried both mutations. The C677 --> T mutation in the methylenetetrahydrofolate reductase gene was assessed in 83 patients; 15 of them (18%) were homozygous and 37 (44%) were heterozygous. There was no significant association between the status of the mutation and history of venous and arterial thrombosis. No significant correlation was found among the three groups. In conclusion, only the G1691 --> A mutation in the factor V gene was associated with the thrombotic risk of patients with lupus anticoagulants. PMID- 10792298 TI - Antibodies to tissue-type plasminogen activator in plasma from patients with primary antiphospholipid syndrome. AB - A reduction in fibrinolysis has been described in association with thrombosis in the primary antiphospholipid syndrome (PAPS). In this study, we measured anti tissue-type plasminogen activator (t-PA) antibodies and anti-fibrin-bound t-PA antibodies as possible causes of hypofibrinolysis in 39 patients with PAPS. We also evaluated the differences in anti t-PA antibodies between patients without previous thrombosis (20 patients) and patients with previous episodes of thrombosis (19 patients: deep vein thrombosis in nine, ischaemic stroke in six, arterial leg thrombosis in one, hepatic vein thrombosis in one, thrombophlebitis in one and cerebral venous thrombosis in one). Anti-t-PA antibodies were measured by an enzyme-linked immunosorbent assay (ELISA), and anti-t-PA fibrin-bound antibodies were measured by a solid-phase fibrin immunoassay (SOFIA) in 39 patients with PAPS and in 39 controls matched for gender and age. High levels of IgG anti-t-PA were found in three out of 39 patients with PAPS, and all three patients had a history of thrombosis; four other patients, one of whom had a history of thrombotic events, had high titres of antibodies directed against fibrin-bound t-PA. In addition, patients with ischaemic stroke had significantly higher levels of IgG anti-t-PA than patients without thrombosis (P = 0.029). In conclusion, our data showed that, in patients with PAPS, the highest levels of anti-t-PA antibodies were present in subjects with previous thrombotic events. The discrepancy in the results obtained with two methods of detection of anti-t PA antibodies, ELISA and SOFIA, indicates a different interaction of the antibodies with the t-PA molecules, which are directly bound to polystyrene plates in ELISA and bound to fibrin as a bridging molecule in SOFIA. PMID- 10792300 TI - Effects of intravenous immunoglobulin in a patient with intermittent thrombotic thrombocytopenic purpura. AB - We describe a patient with a 9-year history of thrombotic thrombocytopenic purpura (TTP) who exhibited four relapses. Intravenous immunoglobulin (IVIg) was effective for these four episodes. The patient was well and the laboratory findings were within normal ranges between each episode, although unusually large von Willebrand factor multimers were observed during remission. Our results suggest the usefulness of IVIg at the time of relapse in the treatment of patients with TTP who have multiple relapses over a long period. PMID- 10792299 TI - Autosomal dominant type 1 von willebrand disease due to G3639T mutation (C1130F) in exon 26 of von Willebrand factor gene: description of five Italian families and evidence for a founder effect. AB - Twenty-four apparently unrelated Italian patients with autosomal dominant type 1 von Willebrand disease (VWD) and a clear autosomal pattern of inheritance of bleeding symptoms were screened for the C1149R and C1130F mutations. None of the patients had the C1149R mutation; three patients and four affected relatives were heterozygous for the C1130F mutation. The mutation appeared to be linked to a single haplotype, defined by five genetic markers [variable number tandem repeat (VNTR) I and II in intron 40, RsaI in exons 13 and 18 and HphI in exon 28], suggesting a founder effect. The patients responded well to desmopressin infusion. The C1130F mutation might have a dominant negative effect on the secretion of the normal protein that desmopressin would appear to overcome. PMID- 10792301 TI - 'Blood doping' with recombinant erythropoietin (rhEPO) and assessment of functional iron deficiency in healthy volunteers. PMID- 10792303 TI - Reply to drach et al PMID- 10792302 TI - Deletions of the p53 gene in multiple myeloma. PMID- 10792304 TI - Barrett's oesophagus--50 years on. PMID- 10792305 TI - On getting started. PMID- 10792306 TI - A bile duct stone cannot be found. PMID- 10792307 TI - Laparoscopic surgery and surgical infection. AB - INTRODUCTION: It has been observed that the metabolic response to surgical injury is less after laparoscopic surgery than after open surgery. However, the effect of laparoscopic surgery on surgical infection has not been given much attention in the surgical literature, even though it may decrease the incidence of infectious complications. The objective of this study was to assess the influence that laparoscopic surgery has on surgical infection and to highlight certain controversial aspects. METHODS: A review of the literature was undertaken to examine the relationship between laparoscopic surgery and surgical infection. This was achieved primarily by using PubMed Medline as a source of material. RESULTS AND CONCLUSION: Laparoscopic surgery is associated with better preservation of the immune system than open surgery. This results in a decreased incidence of infectious complications. Although carbon dioxide pneumoperitoneum affects the peritoneal response to injury, it seems to have no harmful effect in terms of intra-abdominal infection. Nevertheless, at laparoscopic operation the virulence of intestinal micro-organisms should be recognized and, while knowing the advantages of minimally invasive surgery, the surgeon should consider the complexity of this technique. Furthermore, maintenance of laparoscopic instruments should be governed by the same norms as those used in open surgery; recommendations offered by the manufacturers should be respected. PMID- 10792308 TI - Sportsman's hernia. AB - BACKGROUND: Sportsman's hernia is a debilitating condition which presents as chronic groin pain. A tear occurs at the external oblique which may result in an occult hernia. The definition, investigation and treatment of this condition remain unclear. METHODS: A systematic Medline search was performed and all literature pertaining to chronic groin pain, groin injury, sportsman's hernia and sportsman's groin from 1962 to 1999 was retrieved for analysis. RESULTS: The costs of computed tomography and magnetic resonance imaging are such that their routine use for assessment of patients with groin pain cannot be justified. They may, however, be employed in difficult cases to help define the anatomical extent of a groin injury. Plain radiography, ultrasonography and scintigraphy should be the usual first-line investigations to supplement clinical assessment. Herniography may help in situations of obscure chronic groin and pelvic pain. There is no consensus view supporting any particular surgical procedure for sportsman's hernia. A number of reports have been published describing different repairs of the posterior inguinal wall deficiency. Appropriate repair of the posterior wall results in therapeutic benefit in selected cases. CONCLUSION: The diagnosis of sportsman's hernia is difficult. The condition must be distinguished from the more common osteitis pubis and musculotendinous injuries. Early surgical intervention is usually, although not always, successful when conservative management has failed. PMID- 10792310 TI - DIGEST: digest PMID- 10792309 TI - Exercise training and peripheral vascular disease. AB - BACKGROUND: Conservative management is advocated as a treatment of choice for patients with intermittent claudication. This is a review of the mechanisms behind the improvement following an exercise rehabilitation programme. METHODS: All Medline articles from the National Library of Medicine, USA containing the text words 'claudication' or 'peripheral vascular disease' and 'exercise' were reviewed. Cross-referencing from relevant articles was carried out. RESULTS AND CONCLUSION: The poor physical status of a patient with intermittent claudication is not solely due to a reduction in blood flow to the lower limbs; associated factors, such as metabolic inefficiency, poor cardiorespiratory reserve and exercise-induced inflammation contribute. An exercise programme frequently improves both the physical aspect and quality of life, and the success of such exercise is multifactorial. An increase in the blood flow to the lower extremity is uncommon. Other factors, such as a redistribution of blood flow, changes in oxidative capacity of the skeletal muscles and greater utilization of oxygen, occur and the associated metabolic dysfunction of the skeletal muscles is rectified. Following exercise training, blood rheology improves and exercise induced inflammation is ameliorated; cardiorespiratory status also benefits and the oxygen cost of exercise decreases. PMID- 10792311 TI - Effect of liver blood flow and function on hepatic indocyanine green clearance measured directly in a cirrhotic animal model. AB - BACKGROUND: Peripheral blood clearance of indocyanine green (ICG) has been used as a test of liver function but gives little information on biliary excretion. Hepatic ICG uptake and clearance can be measured directly by near-infrared spectroscopy (NIRS). Direct ICG measurement has not previously been correlated with liver blood flow and function in cirrhosis. METHODS: Two groups of New Zealand white rabbits (n = 12) underwent laparotomy for liver exposure. Cirrhosis was induced by feeding animals (n = 6) with a high-cholesterol (2 per cent) diet for 16 weeks. Hepatic blood flow and microcirculation were measured. Hepatic ICG concentration was measured directly using NIRS probes on the liver surface. From the ICG concentration-time curve, hepatic ICG uptake and excretion rates were calculated by a non-linear least square curve fitting method. RESULTS: There was a significant reduction in ICG uptake rate (mean(s.d.) 0.300(0.130) versus 2.040(0.420) min-1; P = 0.0001) and ICG excretion rate (0.007(0.009) versus 0.227(0.096) min-1; P = 0. 002) in cirrhotic animals. The hepatic ICG uptake rate correlated with hepatic blood flow and flow in the microcirculation (r = 0.81, P = 0.002; r = 0.92, P < 0.001, respectively). The hepatic ICG excretion rate was significantly associated with indicators of impaired liver function including bilirubin (r = - 0.86, P = 0.0004), aspartate aminotransferase (r = - 0.81, P = 0.001) and lactate dehydrogenase (r = - 0.83, P = 0.0008). CONCLUSION: ICG uptake measured directly by NIRS reflects the reduced liver blood flow and perfusion in cirrhosis and its excretion correlates with the degree of liver parenchymal dysfunction. This technique may allow a more accurate method of liver function assessment than peripheral blood ICG clearance. PMID- 10792312 TI - Preoperative staging of rectal cancer allows selection of patients for preoperative radiotherapy. AB - BACKGROUND: Variability in rates of local recurrence following resection of rectal cancer has led to the suggestion that all patients should undergo preoperative radiotherapy. This centre employs a selective policy of radiotherapy only in patients with evidence of advanced local disease determined by preoperative staging. METHODS: A retrospective review was carried out of 114 consecutive patients with rectal cancer. Patients were divided before operation into palliative and curative groups based on preoperative staging. Only patients in the palliative group were offered preoperative radiotherapy. Total mesorectal excision (TME) was performed for all tumours of the middle or lower rectum. RESULTS: The perioperative mortality rate was 0.9 per cent and anastomotic dehiscence occurred in 2.8 per cent. Local recurrence developed in 4 per cent of patients in the 'curative' group and in seven of 15 of those assigned to the palliative group before operation (P < 0.01). Positive lateral resection margins were significantly associated with a risk of subsequent recurrence (ten of 13 versus three (3 per cent) of 93; P < 0.001). CONCLUSION: Preoperative adjuvant radiotherapy can be omitted reasonably in patients in whom there is no evidence of locally advanced disease, provided that adequate surgery, incorporating TME for low tumours, is performed. PMID- 10792313 TI - Reduced endothelin-3 expression in sporadic Hirschsprung disease. AB - BACKGROUND: Enteric aganglionosis in Hirschsprung disease has been linked to genes coding for endothelin-3 (EDN3) and the endothelin B receptor (EDNRB), but there is no such linkage in most patients with sporadic Hirschsprung disease. However, the similarity between the distal colonic aganglionosis in Hirschsprung disease and that due to EDN3 or EDNRB mutations led to the hypothesis that levels of expression of these genes might be affected in the absence of mutation, thus causing the Hirschsprung disease phenotype. The aim of this study was to determine EDN3 and EDNRB messenger RNA (mRNA) levels in tissue samples from patients with sporadic Hirschsprung disease. METHODS: RNA and DNA were isolated from the ganglionic and aganglionic colonic segments of ten children with sporadic Hirschsprung disease, and from the colon of ten age-matched controls. The DNA was analysed for mutations in the genes coding for endothelin-3 (ET-3) and endothelin B receptor (ET-B) proteins. Relative levels of EDN3 and EDNRB mRNA were determined by semi-quantitative transcriptase-polymerase chain reaction. RESULTS: Three children had sequence variants in EDN3 and EDNRB. In the remaining seven patients, EDN3 mRNA levels were reduced in both the ganglionic and aganglionic colon compared with levels in controls; there was no difference in expression of EDNRB between groups. CONCLUSION: In the absence of mutation, EDN3 is downregulated in short-segment Hirschsprung disease, suggesting that this may be a common step leading to aganglionosis. PMID- 10792314 TI - Manometric assessment of an artificial bowel sphincter. AB - BACKGROUND: This study investigated the relationship between functional clinical results after artificial bowel sphincter implantation and manometric assessment in 12 consecutive patients. METHODS: A postoperative manometric study was performed in 12 patients, including measurement of resting and squeeze pressures, opening characteristics and pressure during straining. The relationship between postoperative findings, clinical outcome and preoperative manometric data was investigated. Anal pressure after rectal distension with a closed cuff was studied in seven patients. Results were expressed as mean(s.d.). RESULTS: Continence for solid stool was achieved in all 12 patients. Five patients remained incontinent for gas. Anal resting pressure was 108(22) cmH2O; there was no difference between continent and incontinent patients. Rectal distension induced anal relaxation in six of seven patients. Total duration of cuff opening was 113(8) s with an amplitude of 60(22) per cent; residual pressure was correlated with the preoperative resting pressure. The total duration of the opening phase in patients with defaecation difficulties (47(24) (range 0-65) s) was shorter than that in patients without defaecation difficulties (178(78) (range 100-320) s) (P = 0.0022). CONCLUSION: Postoperative defaecation difficulties after implantation of an artificial bowel sphincter are related to a short duration of opening of the cuff. The anal sphincter played a role in postoperative resting anal pressures and allowed relaxation even if the cuff was closed. PMID- 10792315 TI - Quality of life after total colectomy with ileorectal anastomosis or proctocolectomy and ileal pouch-anal anastomosis for familial adenomatous polyposis. AB - BACKGROUND: Knowledge of postoperative health status is important in decision making about the type of operation necessary in patients with familial adenomatous polyposis (FAP). This study compared the quality of life (QoL) between patients with an ileorectal anastomosis (group 1) and those with an ileal pouch-anal anastomosis (group 2). METHODS: QoL was assessed with both a generic questionnaire (Short Form-36 Health Survey; SF-36) and a disease-specific questionnaire (European Organization for Research and Treatment of Cancer Colorectal QoL Questionnaire; EORTC QLQ-CR38). The SF-36 consists of 36 items representing eight generic health domains, and the EORTC QLQ-CR38 comprises 38 items representing disease-specific health domains. Both questionnaires were distributed among 323 patients with FAP known at the Dutch Polyposis Registry who had previously undergone either operation. The results of the SF-36 were compared with the scores of age- and sex-matched respondents from the general population. RESULTS: Some 279 patients (86 per cent), 161 in group 1 and 118 in group 2, completed the questionnaire. Generic and disease-specific QoL was the same for groups 1 and 2. The SF-36 scores of both groups were significantly lower than those of the general population. CONCLUSION: There were no differences with respect to health status between patients in groups 1 and 2, and preference for either procedure cannot be based on QoL. PMID- 10792316 TI - Intraoperative sentinel lymph node examination by imprint cytology and frozen sectioning during breast surgery. AB - BACKGROUND: The purpose of the present study was to evaluate the usefulness of intraoperative imprint cytology and frozen sectioning of sentinel lymph nodes in patients with clinically node-negative breast cancer. METHODS: Sentinel node biopsy was performed in 101 patients with stage I or II breast cancer with clinically negative nodes using a dye-guided method. Intraoperative evaluation of sentinel node involvement by imprint cytology and frozen sectioning was compared with the final histopathological results of permanent sections. Tumour-negative nodes in paraffin sections stained by haematoxylin and eosin were further studied using an anticytokeratin antibody. RESULTS: The results of imprint cytology and frozen-section analysis were compared with those of haematoxylin and eosin stained sections. The sensitivity, specificity and overall accuracy of imprint cytology were 96.0, 90.8 and 92.1 per cent respectively, and those of frozen section examination were 52.0, 100 and 88.1 per cent. Ten sentinel nodes were tumour positive on imprint cytology and tumour negative on stained paraffin sections. Micrometastasis was found in eight of these nodes on immunohistochemistry. Taking these immunohistological results into consideration, the final sensitivity, specificity and overall accuracy of imprint cytology were 90.9, 98.5 and 96.0 per cent respectively. CONCLUSION: Intraoperative imprint cytology is a useful method for evaluating the status of sentinel nodes and is more accurate than frozen-section analysis. In addition, imprint cytology can detect micrometastasis more accurately than conventional haematoxylin and eosin stained sectioning. PMID- 10792317 TI - Gunshot injuries of the popliteal artery. AB - BACKGROUND: Guidelines for the management of popliteal artery trauma emanate mainly from military experience. This study was undertaken to describe the management of popliteal injuries in a civilian vascular surgical unit with a large trauma workload. METHODS: A retrospective review of records of patients treated between 1983 and 1997 was undertaken. RESULTS: Some 117 popliteal artery gunshot injuries were treated (83 low velocity, 16 high velocity, 18 shotgun). Associated fractures occurred in 44 patients and 40 had popliteal vein injuries. Treatment of the arterial injury included vein graft interposition in 71, primary reanastomosis in 19, prosthetic graft interposition in four, lateral suture in one, vein patch in one and ligation in one patient; 84 fasciotomies were performed. No perioperative deaths occurred. There were 20 primary and 14 secondary amputations. Factors associated with amputation were high-velocity injuries, delay in revascularization in excess of 7 h, arterial transection, associated fracture, and compartment syndrome or muscle infarction. CONCLUSION: Civilian popliteal gunshot injuries are attended by a high amputation rate. Prompt resuscitation and revascularization appear to be the only correctable factors that may improve limb salvage rates. PMID- 10792319 TI - Diagnostic failures in endoscopy for acute upper gastrointestinal haemorrhage. AB - BACKGROUND: Oesophagogastroduodenoscopy (OGD) is the diagnostic tool of choice in acute upper gastrointestinal haemorrhage. However, the factors causing diagnostic failure are not well documented or discussed. METHODS: OGDs performed by a single surgeon for acute upper gastrointestinal haemorrhage were reviewed retrospectively for 'missed' diagnosis. RESULTS: There were nine cases (1.4 per cent) of 'missed' diagnosis in 638 OGDs performed over a 3-year period. Incomplete examination caused by a fundal pool precluded three examinations in which two Dieulafoy's lesions and one chronic gastric ulcer were later found. Other difficult examination sites were the cardia/high lesser curve (three cases), the stomal line of a gastrojejunostomy anastomosis (two) and at the junction of the first and second part of the duodenum (one). Four of the overlooked diagnoses were Dieulafoy's lesions. CONCLUSION: OGD was able to diagnose the cause of bleeding in 98.6 per cent of patients with acute upper gastrointestinal haemorrhage. The factors that caused diagnostic failure were a difficult examination site and failure to recognize lesions (Dieulafoy's). PMID- 10792318 TI - Surgical management of intra-abdominal desmoid tumours. AB - BACKGROUND: Intra-abdominal desmoids are uncommon neoplasms. The aggressive nature of these tumours and the potential for major morbidity secondary to resection can present a difficult surgical dilemma. METHODS: Patients with histologically confirmed intra-abdominal desmoid tumours undergoing laparotomy were identified from a prospective database. Clinical features and outcomes in this group were evaluated. RESULTS: The study group comprised 24 patients. Sixteen patients underwent complete resection of the tumour while eight had biopsy only, with or without intestinal bypass. Small intestinal resection was performed in 12 patients, including three who had a near-total enterectomy. Median follow-up was 62 months, with an actuarial overall survival rate of 73 per cent at 10 years. There was no difference in survival rate between completely resected and unresected patients (P = 0.73). There were seven deaths in the entire group, of which four were in those undergoing complete resection. CONCLUSION: Operation can cure patients with intra-abdominal desmoid tumours, but may result in significant morbidity, especially from loss of small intestine. No other therapy is a predictably good alternative to operation but the natural history of desmoids is often characterized by prolonged periods of stability or even regression. A period of watchful waiting, until significant symptoms develop, may be the most appropriate course in patients who risk mesenteric vascular injury or substantial enterectomy with attempts at resection. PMID- 10792320 TI - Changing pattern of histological type, location, stage and outcome of surgical treatment of gastric carcinoma. AB - BACKGROUND: There are indications that some features of gastric carcinoma are changing, with a possible impact on prognosis. The aim of this study was to examine any changes in type, location, stage, resection rate, postoperative mortality rate or prognosis for patients with gastric carcinoma in a well defined population. METHODS: During 1974-1991, 1161 new cases of gastric adenocarcinoma were diagnosed in Ostergotland County, Sweden. Tumour location, Lauren histological type, tumour node metastasis (TNM) stage, radicality of tumour resection and postoperative complications were recorded after histological re evaluation of tissue specimens and examination of all patient records. Dates of death were obtained from the Swedish Central Bureau of Statistics. Time trends were studied by comparing the intervals 1974-1982 (period 1) and 1983-1991 (period 2). RESULTS: The proportion of diffuse type of adenocarcinoma increased (from 27 to 35 per cent), while that of mixed type decreased (from 16 to 9 per cent) and that of intestinal type was unchanged. The proportion of tumours located in the proximal two-thirds of the stomach increased (from 32 to 42 per cent) and the proportion of patients with tumours in TNM stage IV decreased (from 32 to 25 per cent). Overall tumour resection rates were unchanged, although the proportion of radical total gastrectomies increased (from 36 to 50 per cent). Excluding tumours of the cardia or gastric remnant after previous ulcer surgery, the 5-year relative survival rate after radical resection increased from 25 to 36 per cent and the postoperative mortality rate decreased for both radical (from 11 to 4 per cent) and palliative (from 18 to 6 per cent) resection. CONCLUSION: The patterns of tumour histology, location and stage of gastric carcinoma have changed in the authors' region. These changes were paralleled by a significant improvement in survival and postoperative mortality rates. PMID- 10792321 TI - Prognosis after breast recurrence following conservative surgery and radiotherapy in patients with node-negative breast cancer. PMID- 10792322 TI - Authors' reply PMID- 10792323 TI - Endoscopic technique for the localization of intraduct papillomas. PMID- 10792324 TI - Author's reply PMID- 10792325 TI - Endoscopically assisted, minimally invasive parathyroidectomy. PMID- 10792326 TI - Authors' reply PMID- 10792327 TI - Epidemiology of Warthin's tumour of the parotid gland in an Asian population. PMID- 10792328 TI - Prospective study of chronic pain after groin hernia repair. PMID- 10792329 TI - What's best for the bones in Turner syndrome? PMID- 10792330 TI - A longitudinal study on bone mineral density until adulthood in girls with Turner's syndrome participating in a growth hormone injection frequency-response trial. AB - OBJECTIVE: The aim of this study was to assess the volumetric bone mineral density (BMD) in girls with Turner's syndrome (TS) before and during growth hormone (GH) treatment in combination with low dose oestrogens as well as three years after discontinuation of GH treatment. DESIGN: In a prospective, randomized injection frequency-response study, the effect of GH treatment in combination with low dose ethinyl oestradiol (starting with 0.05 microgram/kg/day), on BMD was evaluated, comparing twice daily (BID) with once daily (OD) injections of a total GH dose of 6 IU/m2/day until adult height was reached. After discontinuation of GH treatment, the dosage of oestrogens was further increased to adult supplementation levels. PATIENTS: Nineteen untreated girls with TS, mean (SD) baseline pretreatment age 13.3 (1.7) (range 11.0-17.6) year. MEASUREMENTS: Before and during GH treatment, measurements of volumetric BMD were performed using phalangeal radiographic absorptiometry. In addition, the BMD measurements were repeated three years after discontinuation of GH treatment. BMD results were adjusted for bone age and sex, and expressed as SD-scores (SDS) using reference values of healthy Dutch girls. RESULTS: At baseline, most individual BMD values of cortical bone as well as those of trabecular bone were within the normal range of healthy girls. However, the mean BMD SDS of the trabecular bone was significantly lower than zero. During treatment, the BMD SDS showed a significant increment to values equal or higher than zero after mean (SD) GH treatment period of 36.6 (7.5) months. The increase in BMD of the cortical bone was significantly higher in the OD group than in the BID group. The BMD SDS in the last year of GH treatment was not significant different between the two injection frequency groups. Three years after discontinuation of GH treatment, the BMD values had increased further similar as in healthy girls, resulting in BMD values all within normal range or even higher. CONCLUSIONS: Most untreated girls with Turner syndrome, age >/= 11 years, have a normal volumetric BMD of the cortical, as well as of the trabecular bone compared to healthy girls. During GH treatment with 6 IU/m2/day in combination with low dose oestrogens, the BMD SDS increases significantly. After discontinuation of GH treatment and the use of oestrogens in an adult dosage, the BMD was as high as in young healthy women. PMID- 10792331 TI - Pre-treatment IGF-I level is the major determinant of GH dosage in adult GH deficiency. AB - OBJECTIVE: Severe GH deficiency in adults is a definite clinical entity, the effects of which can be reversed by administration of subcutaneous recombinant GH. The ideal dosing regimen and determinants of the maintenance dose have, however, yet to be elucidated. PATIENTS: In an open study of GH replacement we treated 65 GH-deficient adults of mixed adult- and childhood-onset, of mean age 35.5 (range 17-72) years, and comprising 38 females and 27 males, using an individualized low-dose titration regimen aimed at normalization of the serum IGF I and induction of clinical improvement. RESULTS: Before initiation of GH therapy, median IGF-I SD was significantly lower in female than male patients (- 3.3 vs. - 1.9, P = 0.007) and in childhood-onset compared with adult-onset patients (- 3.9 vs. - 2.0, P < 0.001). Once maintenance dosage had been achieved, the median GH requirement was significantly greater in female than male patients (1.6 vs. 0.8 IU/day, P = 0.013) and childhood-onset compared with adult-onset patients (1.6 vs. 0.8 IU/day, P = 0.019). The median maintenance GH dose for the cohort overall was 1.2 (range 0.4-2.4) IU/day. By univariate analysis a significant negative correlation was observed between the maintenance GH dose and baseline IGF-I SD (r = - 0.63, P < 0.001). No significant correlation was demonstrated between maintenance GH dose and either age or weight. Multiple linear regression analysis using age, gender, weight, time of onset of GH deficiency, peak GH to the insulin tolerance test (ITT) and baseline IGF-I SD as independent variables demonstrated baseline IGF-I SD to account for 51% of the variation in GH dose required to normalize the IGF-I SD (P < 0.001). Those patients with the lower IGF-I SD at initiation of GH therapy required the greater GH dose. None of the other variables studied significantly influenced the maintenance dose. CONCLUSION: We have demonstrated that the GH dose required in an individual is dependent on the serum IGF-I SD before commencement of replacement. In contrast, the severity of GH deficiency as judged by the peak GH response to an ITT was unrelated to the maintenance GH requirement. The effect of age, gender and age at onset of GH deficiency on the final GH dose are accounted for by the lower pretreatment IGF-I SD in young, female and childhood-onset patients relative to older, male and adult-onset patients, respectively. PMID- 10792332 TI - Increase of adhesion molecules, fibrinogen, type-1 plasminogen activator inhibitor and orosomucoid in growth hormone (GH) deficient adults and their modulation by recombinant human GH replacement. AB - OBJECTIVE: GH deficiency (GHD) is usually associated with a higher incidence of cardiovascular disease (CVD). The aim of this study was to establish whether patients with GHD, like those with CVD, show an increase in fibrinogen (FBG), type-1 plasminogen activator inhibitor (PAI-1), acute phase response proteins (APR), and soluble adhesion molecules. The effect of recombinant human GH (rhGH) replacement, on these parameters was also investigated. PATIENTS AND DESIGN: Concentrations of PAI-1 antigen (Ag), adhesion molecules sE-selectin, sP selectin, and intercellular adhesion molecule-1 (sICAM-1), FBG and levels of APR orosomucoid (ORM), and 'negative APR' transferrin (TRF) were established in 11 panhypopituitary (PHP) patients (eight men and three women, age median 39.0 years, body mass index (BMI) 27. 49 +/- 3.89 kg/m2) before and after 12-month replacement with rhGH. Control values were obtained by examination of 33 healthy age and sex matched subjects (24 men and nine women with BMI 24.16 +/- 1.99 kg/m2). RESULTS: PHP patients had higher concentrations of ORM (0.80 +/- 0.25, vs. 0.61 +/- 0.20 g/l; P = 0.05), FBG (3.22 +/- 0.48, vs. 2.57 +/- 0.47 g/l; P = 0.001), PAI-1 Ag (97.12 +/- 33.23, vs. 44.11 +/- 21.40 microgram/l; P = 0.001), sE-selectin (72.42 +/- 28.35, vs. 42. 80 +/- 12.60 microgram/l; P = 0.004), sP selectin (221.26 +/- 75.12, vs. 104.79 +/- 26.01 microgram/l; P = 0.001) sICAM-1 (409.75 +/- 137.78, vs. 228.10 +/- 37.54 microgram/l; P = 0.001), and lower levels of TRF (2.14 +/- 0.40, vs. 2.76 +/- 0.39 g/l; P = 0.001) than controls. After 12-month rhGH replacement the patients showed an increase of TRF (2.64 +/- 0.84 g/l, P = 0.037) and decrease of soluble adhesion molecules (sE-selectin 57.98 +/- 27.04 microgram/l, P = 0.01, sP-selectin 121.74 +/- 50.42 microgram/l, P = 0.007; and sICAM-1 279.95 +/- 88.32 microgram/l, P = 0.005), which then, similarly to the ORM (0. 67 +/- 0.12 g/l) and FBG level (2.82 +/- 0.51 g/l), did not statistically differ from the values in the control group. CONCLUSION: rhGH replacement led to modulation of the 'inflammatory response' in panhypopituitary patients. This modulation occurred locally at vascular endothelium level where after rhGH replacement, sE-selectin, sP-selectin and sICAM-1 concentrations decreased, a similar effect as in the systemic inflammatory response, as was also apparent from the changes in acute phase response protein levels. PMID- 10792333 TI - Impaired beta-cell function in the presence of reduced insulin sensitivity determines glucose tolerance status in acromegalic patients. AB - OBJECTIVE: Abnormal glucose tolerance is often demonstrated in acromegalic patients. Although insulin resistance is a common feature of acromegaly, it remains unclear whether the extent of insulin resistance per se determines the abnormal glucose tolerance. In order to elucidate this issue, we investigated insulin sensitivity and beta-cell function in acromegalic patients. DESIGN: Twenty-four acromegalic patients were studied in comparison with 24 healthy control subjects. To estimate insulin sensitivity and beta-cell function, we used correct homeostasis model assessment (HOMA) model, a computer-solved model. We also investigated the effects of surgical success on both parameters. RESULTS: HOMA insulin sensitivity (HOMA-%S) in the acromegalic patients was 74 +/- 51 (SD)%, significantly lower than that in 24 healthy controls (144 +/- 49%). HOMA %S in 12 normal glucose tolerance (NGT) patients was 54 +/- 31%, not significantly different from that in impaired glucose tolerance (IGT; n = 11) or diabetes mellitus (DM; n = 1) patients (93 +/- 60%). By contrast, HOMA beta-cell function (HOMA-%beta) in the NGT acromegalic patients was 163 +/- 67%, significantly higher than the IGT/DM acromegalic patients (89 +/- 34%) and the healthy controls (72 +/- 19%). In 11 patients who achieved complete normalization of GH excess after surgery, HOMA-%S significantly increased to control ranges (from 76 +/- 26 to 159 +/- 61%) within 2 weeks after the surgical success. CONCLUSIONS: We conclude that insulin sensitivity is reduced to a similar extent in acromegalic patients with normal glucose tolerance and those with impaired glucose tolerance or diabetes. Compensatory hyperfunction of beta-cells appears to counterbalance the reduced insulin sensitivity in the acromegalic patients with normal glucose tolerance but not in those with impaired glucose tolerance or diabetes. PMID- 10792334 TI - Mitochondrial DNA A3243G mutation in patients with early- or late-onset type 2 diabetes mellitus in Hong Kong Chinese. AB - BACKGROUND AND OBJECTIVES: The mitochondrial DNA A to G mutation at nucleotide 3243 (mt3243) is associated with a subtype of diabetes characterized by maternal transmission and deafness. We have previously reported a 2.7% prevalence of this mutation in a cohort of young patients with either type 1 or type 2 diabetes. In this study, we aimed to confirm this finding by examining for the prevalence of this mutation in a large-scale study. SUBJECTS AND METHODS: Nine hundred and six unrelated Chinese patients with type 2 diabetes and 213 nondiabetic controls were studied. The presence of mt3243 mutation was determined by polymerase chain reaction amplification and ApaI digestion. RESULTS: This mutation was found in four of 133 (3.0%) patients with early onset ( 40 years) diabetes and no family history. Basal pancreatic beta-cell function, as assessed by fasting plasma C-peptide, was variable amongst mutation carriers, and did not correlate with the level of heteroplasmy of mutation. CONCLUSIONS: In agreement with most studies, our results suggest that despite the high prevalence of positive maternal family history of diabetes amongst our type 2 diabetic patients, mt3243 mutation was not a major cause of diabetes in either early- or late-onset diabetic patients in Hong Kong. The role of other genetic, environmental and intrauterine factors needs further investigation. PMID- 10792335 TI - Increased risk for endocrine autoimmunity in Italian type 2 diabetic patients with GAD65 autoantibodies. AB - OBJECTIVE: Glutamic acid decarboxylase (GAD)65 autoantibodies (GAD65Ab) in type 2 diabetic subjects with secondary failure to sulphonylurea treatment identify the so-called latent autoimmune diabetes of the adult (LADA). The aim of our study was to estimate the risk for endocrine autoimmunity in type 2 diabetic subjects with GAD65Ab. DESIGN AND PATIENTS: We analysed serum samples from 600 adult subjects with a clinical diagnosis of type 2 diabetes mellitus for the presence and levels of GAD65Ab and antibodies directed against the islet autoantigen IA 2/ICA512 (IA-2/ICA512Ab). All the patients had been treated initially with hypoglycaemic agents and/or diet for at least 1 year. GAD65Ab+ subjects were studied for the presence of thyroid peroxidase autoantibodies (TPOAb), 21 hydroxylase autoantibodies (21OHAb) and frequency of HLA class II haplotypes. RESULTS: GAD65Ab were found in 67/600 (11%) and IA-2/ICA512Ab in 12/600 (2%) subjects (P < 0.0001). The presence of GAD65Ab, but not that of IA-2/ICA512Ab, was significantly associated with insulin therapy, low BMI (P < 0.0001) and low basal C-peptide (P < 0.01). Islet-cell antibodies (ICA) were detected in 43/67 (64%) GAD65Ab+ and in 10/12 (83%) IA-2/ICA512Ab + subjects. TPOAb occurred more frequently in GAD65Ab+ (16/67, 24%) than in GAD65Ab-subjects (9/174, 5%) (P < 0.0001). 21OHAb were detected only in GAD65Ab+ subjects (3/67, 4.5%) (P = 0.03 vs. GAD65Ab-subjects). None of the 21OHAb+ subjects had metabolic or clinical signs of adrenal dysfunction. HLA-DRB1*03-DQA1*0501-DQB1*0201 (DR3-DQ2) was significantly more frequent in GAD65Ab+ subjects than in healthy controls (OR = 5.42, corrected P < 0.0026). The presence of TPOAb was significantly associated with DR3-DQ2 (P = 0.024). CONCLUSIONS: Our study demonstrates that the presence of GAD65Ab identifies a subgroup of type 2 diabetic patients with high risk for thyroid and adrenal autoimmunity, and that both GAD65Ab and TPOAb are associated with the presence of HLA-DR3-DQ2, in these patients. PMID- 10792336 TI - Vitamin D receptor gene polymorphisms influence susceptibility to type 1 diabetes mellitus in the Taiwanese population. AB - OBJECTIVE: Vitamin D and its receptor have been suggested to play a role in the pathogenesis of type 1 diabetes mellitus. We have therefore studied the influence of vitamin D receptor (VDR) gene polymorphisms on susceptibility to type 1 diabetes, and rates of glutamic acid decarboxylase (GAD65) autoantibody and islet cell autoantibody (ICA512) positivity. SUBJECTS: AND MEASUREMENTS One hundred and fifty-seven type 1 diabetic patients and 248 unrelated normal controls were recruited for this study. Genomic DNA was extracted from peripheral blood leucocytes. All type 1 diabetic patients and controls were genotyped using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP), for three restriction sites in the VDR gene, BsmI, ApaI and TaqI. The chi2 test was used to compare the frequency of the VDR gene polymorphisms in patients and normal controls. The association of VDR gene polymorphisms in type 1 diabetes with the presence of GAD65 and ICA512 autoantibodies were also examined using the chi2 test. RESULTS: The allele frequency of the BsmI and ApaI polymorphisms, but not TaqI polymorphism, differed between patients and controls (BsmI P = 0.015; ApaI P = 0.018; TaqI P = 0.266). However, after correction for the three different polymorphisms tested, only the BsmI was significant (pc = 0.045). CONCLUSIONS: Vitamin D receptor gene polymorphisms were associated with type 1 diabetes in a Taiwanese population. However, functional studies are needed to establish the role of the vitamin D receptor in the pathogenesis of type 1 diabetes mellitus. PMID- 10792337 TI - Oestrogen-receptor-alpha gene polymorphism affects response in bone mineral density to oestrogen in post-menopausal women. AB - OBJECTIVE: An oestrogen-receptor-alpha (ERalpha) gene polymorphism has been variably reported to be related to bone mass. To investigate whether this ERalpha gene polymorphism is associated with a functional difference, we assessed the response in bone mineral density (BMD) to oestrogen therapy in post-menopausal women in relation to ERalpha gene polymorphism. PATIENTS AND MEASUREMENTS: Subjects consisted of 124 Thai post-menopausal women. Sixty-three of the women were less than 6 years post-menopausal and 61 were more than 10 years post menopausal with vertebral or femoral osteoporosis as defined by BMD T-score less than - 2.5. Subjects were randomly allocated to receive 0.3 mg (n = 67) or 0.625 mg (n = 57) of conjugated equine oestrogen (CEE). All subjects also took 5 mg medroxyprogesterone acetate. Vertebral and femoral neck BMD were measured at baseline and 1 year after treatment. Data were expressed as mean +/- SEM. Capital P represents the absence of the restriction site while lower-case p indicates the presence of the restriction site. RESULTS: For subjects on 0.625 mg CEE, BMD at L2-4 increased significantly after 1 year in those with pp (n = 20) Pp (n = 29) and PP genotypes (n = 8) (P < 0.001). However, in subjects on 0.3 mg CEE, BMD at L2-4 increased significantly after 1 year in subjects with Pp (n = 34, + 7.6 +/- 1.5%, P < 0.001) and PP genotypes (n = 13, + 6. 9 +/- 1.0%, P < 0.001), but not in those with pp genotype (n = 20, + 2.3 +/- 2.1%, P = NS). After adjusting for age and years since menopause, the change in vertebral BMD was still lower in those without the P allele compared to those with the P allele (P < 0.05). Femoral BMD did not significantly change regardless of dose of CEE and genotype. CONCLUSIONS: We conclude that ERalpha gene polymorphism affects skeletal response to oestrogen in post-menopausal women. The effect of ERalpha gene polymorphism appears to be site-specific and does not relate to biochemical markers of bone turnover. Determination of ERalpha genotype may help identify post-menopausal women who will have more skeletal benefit from oestrogen therapy. PMID- 10792338 TI - Spironolactone as a single agent for long-term therapy of hirsute patients. AB - OBJECTIVE: To assess the androgen-suppressing effect of spironolactone, and the use of this drug as a single agent in the long-term therapy of hirsute patients with either polycystic ovary syndrome (PCOS) or idiopathic hirsutism (IH). Standard cyproterone acetate (CPA) treatment was used to evaluate the results obtained with spironolactone. DESIGN: Prospective randomized study. PATIENTS: Forty-six hirsute women were separated randomly into two groups, stratified for polycystic ovary syndrome. For 12 months, Group 1 (21 patients, 10 PCOS) received spironolactone only (200 mg/day). Group 2 (23 patients, nine PCOS) received CPA (50 mg/day) with ethinyl oestradiol (35 microgram/day). MEASUREMENTS: Ferriman Gallwey clinical score for hirsutism and serum testosterone, androstenedione, and LH levels. RESULTS: In IH patients, hirsutism regressed equally with spironolactone (21 +/- 2-14.5 +/- 2) and CPA (23 +/- 2-13 +/- 2). In PCOS patients, the mean score for hirsutism after 12 months was significantly lower with CPA (12 +/- 1) than with spironolactone (16 +/- 1). Testosterone levels did not change with spironolactone; with CPA there was a decrease from baseline in PCOS (47% and 51%, 6 and 12 months) and IH patients (31% and 30%). Androstenedione levels also declined from baseline in CPA-treated PCOS patients (38% and 39%, 6 and 12 months). Androgen levels were significantly different between the groups after 6 and 12 months. LH levels decreased with CPA (72%) but not with spironolactone. CONCLUSION: Our results suggest that spironolactone used as a single agent is as effective as cyproterone acetate combined with oestradiol for long-term treatment of patients with idiopathic hirsutism. In PCOS patients, spironolactone is still effective for reducing hirsutism; however, for treatment of the hormonal or metabolic manifestations associated with PCOS, it may be necessary to combine spironolactone with either an antigonadotrophic agent or a drug that improves peripheral insulin sensitivity. PMID- 10792339 TI - Cardiovascular disease in women with polycystic ovary syndrome at long-term follow-up: a retrospective cohort study. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) is associated with higher prevalence of cardiovascular risk factors but the relative prevalence of cardiovascular disease in women with PCOS has not previously been reported. We have compared cardiovascular mortality and morbidity in middle-aged women previously diagnosed with PCOS and age-matched control women. DESIGN: A retrospective cohort study of women diagnosed with PCOS in the United Kingdom before 1979. PATIENTS: Seventy cohort members died before 31 March 1999. Morbidity data were collected from 319 women with PCOS and 1060 age-matched control women. Sixty-one women with PCOS and 63 control women attended a clinical examination. MEASUREMENTS: Data were collected from death certificates, general practitioners' records and questionnaires with measurement of cardiovascular risk factors in a subsample of questionnaire respondents. RESULTS: All-cause and cardiovascular mortality in the cohort were similar to women in the general population (standardized mortality ratios (95% CI): 93 (72-117) and 78 (45-124), respectively). Women with PCOS had higher levels of several cardiovascular risk factors: diabetes (P = 0.002) hypertension (P = 0.04), hypercholesterolaemia (P < 0.001), hypertriglyceridaemia (P = 0.02) and increased waist:hip ratio (P = 0.004). After adjustment for BMI, odds ratios (OR) were 2.2 (0.9-5. 2) for diabetes, 1.4 (0.9-2.0) for hypertension and 3.2 (1.7-6.0) for hypercholesterolaemia. A history of coronary heart disease (CHD) was not significantly more common in women with PCOS (crude OR (95%CI) 1.5 (0.7-2.9)) but the crude OR for cerebrovascular disease was 2.8 (1.1-7.1). CONCLUSION: At long-term follow-up, a history of nonfatal cerebrovascular disease and cardiovascular risk factors including diabetes are more prevalent among women with polycystic ovary syndrome. Morbidity and mortality from of coronary heart disease among women with polycystic ovary syndrome is not as high as previously predicted. This finding challenges our understanding of the aetiology of coronary heart disease in women. PMID- 10792340 TI - Influence of different genotypes on 17-hydroxyprogesterone levels in patients with nonclassical congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - OBJECTIVE: The diagnosis of the nonclassical form of 21-hydroxylase (NC-21OH) deficiency, established before molecular studies, is based on basal 17OH progesterone (17OH-P) values > 15 nmol/l or ACTH-stimulated 17OH-P values > 30 nmol/l. This disease is caused by mutations in the structural gene that can be grouped into three categories: A, B and C, according to the predicted level of enzymatic activity. So, the genotype of the nonclassical form is a combination of mutations that cause moderate impairment of enzymatic activity in one allele and mutations which cause total (A), severe (B: 3%) or moderate (C: 20-60%) impairment of enzymatic activity in the other allele. DESIGN: We analysed the influence of the different genotypes on 17OH-P levels in 58 patients with the nonclassical form of 21OH deficiency. RESULTS: After screening for 18 mutations through Southern blotting, allele-specific polymerase chain reaction (PCR) and enzyme restriction, mutations were identified in 73% of the alleles. Patients with mutations identified in both alleles were divided into groups A/C (n = 18), B/C (n = 3) and C/C (n = 15). The basal and ACTH-stimulated 17OH-P levels in patients with A/C genotype ranged from 1.2 to 153 and 72-363 nmol/l, and in C/C genotype ranged from 0.9 to 72 and 51-363 nmol/l, respectively (P < 0.05 for stimulated levels). The lowest value of ACTH-stimulated 17OH-P levels in fully genotyped patients was 51 nmol/l. Patients with the A/C genotype presented androgen excess symptoms earlier than patients with the C/C genotype. CONCLUSIONS: These data suggest an influence of genotype on phenotype and on 17OH P levels. The high frequency of unidentified mutant alleles in nonclassical 21 hydroxylase deficiency suggests that ACTH-stimulated values of 17OH-P between 30 and 51 nmol/l have overestimated this diagnosis. Genotyping more patients with nonclassical 21-hydroxylase deficiency will help to redefine the cut-off value for ACTH-stimulated 17OH-P for correct diagnosis of this disease. PMID- 10792341 TI - The impact of mild Leydig cell dysfunction following cytotoxic chemotherapy on bone mineral density (BMD) and body composition. AB - BACKGROUND: Overt testosterone deficiency is associated with a reduction in BMD and alteration in body composition. However, there are few data concerning the impact of mild hypogonadism on these parameters. PATIENTS AND METHOD: We have identified a cohort of 36 men aged < 55 years with mild Leydig cell impairment, defined by a raised LH level (LH >/= 8 IU/l) in the presence of a testosterone level in the lower half of the normal range or frankly subnormal (< 20 nmol/l), following treatment with procarbazine-containing chemotherapy regimens or high dose chemotherapy for haematological malignancy. These men underwent measurements of BMD (measured by dual-energy X-ray absorptiometry (DXA), single energy X-ray absorptiometry (SXA) and quantitative CT (QCT)), body composition (DXA), markers of bone turnover, serum lipids and serum IGF-1. To allow for changes that may be directly attributable to the underlying disease or its treatment, results were compared with those obtained in 14 men who had received the same chemotherapy for the same diseases but had normal LH and testosterone levels (controls). RESULTS: When data from all 50 men were considered together there were significant reductions in BMD of the lumbar spine both by DXA (Z = - 0.34, P = 0.01) and QCT (Z = - 1.5, P < 0. 0001), at the femoral neck (Z = - 0.52, P < 0.0001) and distal forearm (Z = - 0.21, P = 0.05). Mean femoral neck BMD was significantly lower in patients compared with controls (Z = - 0.68 vs. Z = - 0.11, P = 0.05) and there was a nonsignificant trend towards lower lumbar spine BMD measured by QCT (Z = - 1.64 vs. Z = - 1.10; P = 0.09). In addition, serum testosterone level and testosterone:LH ratio significantly correlated with femoral neck BMD (r = 0.28, P = 0.05 and r = 0.37, P = 0.008, respectively). There were no significant differences in lean body mass, fat mass and percentage fat between the patients and controls. There was, however, a difference in the distribution of body fat with a propensity for the patients to accrue truncal fat, and the serum testosterone level significantly inversely correlated with percentage of truncal fat (r = - 0.29, P = 0.04). There were no significant differences in lipid levels, IGF-1 levels or markers of bone turnover between the patients and controls. CONCLUSIONS: These data suggest that mild Leydig cell impairment may have significant effects on bone mineral density and may result in subtle body composition changes, although in men who have received cytotoxic chemotherapy, other factors also contribute to the observed osteopenia. Testosterone replacement may be beneficial in some of these men and this requires further evaluation. PMID- 10792342 TI - Oral glucose tolerance testing but not intravenous glucose administration uncovers hyper-responsiveness of hypothalamo-pituitary-adrenal axis in patients with adrenal incidentalomas. AB - OBJECTIVE: Modern imaging techniques are detecting adrenal incidentalomas with increasing frequency. Recent data suggests food-dependent hypercortisolism in a subgroup of patients with bilateral macronodular hyperplasia due to aberrant adrenal responsiveness to gastric inhibitory polypeptide (GIP). We studied the putative influence of food intake on the hypothalamo-pituitary-adrenal (HPA) axis in patients with adrenal incidentalomas and possible mediation by GIP. PATIENTS AND MEASUREMENTS: We examined 15 mildly obese patients with adrenal incidentalomas, eight healthy, lean subjects, and seven obese patients with the metabolic syndrome, who were matched for body weight and age. Each individual underwent oral glucose tolerance testing (OGTT, 75 g glucose), i.v. glucose administration (IVGTT, 30 g glucose over 1 h) and i.v. glucose plus GIP infusion (body weight adapted leading to physiological postprandial GIP serum levels) on three occasions. Plasma glucose, ACTH and cortisol were measured from blood samples taken every 15 minutes from time - 30 minutes to + 75 minutes. RESULTS: OGTT, i.v. glucose administration and GIP infusion led to comparable glucose values within the groups. In contrast to normal subjects and patients with the metabolic syndrome, patients with adrenal incidentalomas had significantly higher mean cortisol values after oral glucose intake as compared to i.v. glucose administration or GIP infusion. The increase in cortisol levels was preceded by a corresponding ACTH increase. No significant effect of GIP administration on cortisol or on ACTH secretion could be detected. CONCLUSIONS: Patients with adrenal incidentalomas showed an abnormal responsiveness of the pituitary-adrenal axis to oral glucose administration. The cortisol peaks in these patients seemed to be ACTH-mediated and were not induced by GIP. PMID- 10792344 TI - Low-dose (1 microgram) adrenocorticotrophin (ACTH) stimulation as a screening test for impaired hypothalamo-pituitary-adrenal axis function: sensitivity, specificity and accuracy in comparison with the high-dose (250 microgram) test. AB - OBJECTIVE: We have shown previously that in contrast to the standard high-dose 250-microgram ACTH test, a low-dose 1-microgram ACTH stimulation test correctly identified all patients with pituitary disease who had impaired hypothalamo pituitary-adrenal (HPA) function. In this study we further compared the performances of these two tests as screening procedures for possible HPA impairment. DESIGN: A comparison of the 1-microgram and the 250-microgram ACTH stimulation tests in healthy controls and in patients with pituitary disease whose HPA axis status was characterized formally by a gold standard test. SUBJECTS: A total of 89 subjects were investigated: 27 healthy normal controls, 43 patients with pituitary disease and normal HPA function, and 19 patients with various pituitary diseases and impaired HPA function. MEASURES: All 89 subjects underwent stimulation with 1 microgram ACTH; 80 also underwent the high-dose 250 microgram ACTH test. A receiver operating characteristic analysis (ROC) was performed to compare the tests. RESULTS: Using a stimulated cortisol > 500 nmol/l as the criterion for a normal response, the 1-microgram ACTH stimulation identified 18 of the 19 subjects with impaired HPA function (94.7% sensitivity with a likelihood ratio of 0.0588 for a negative test). In contrast, 15/16 passed the high-dose test (a 6.2% sensitivity with a likelihood ratio of 0.875 for a negative test). All normal controls, and 36/43 patients with preserved HPA function, passed the 1-microgram ACTH test (90% specificity). This degree of accuracy was unrivalled by the high dose test at all the cut-off levels considered. CONCLUSIONS: More sensitive and accurate, the low-dose 1-microgram ACTH test is as simple and safe as the standard 250-microgram test. We suggest it should replace it in screening for adrenal insufficiency. PMID- 10792343 TI - Growth hormone and prolactin are secreted more irregularly in patients with Cushing's disease. AB - OBJECTIVE: To investigate whether the spontaneous secretion of growth hormone and prolactin in adult patients with pituitary-dependent Cushing's disease is decreased. PATIENTS: Fourteen adult patients (9 women, 5 men; age: 34 +/- 3.4 years, mean +/- SEM) with pituitary-dependent Cushing's disease and 14 controls matched for age, gender and body mass index were studied. METHODS: Blood samples were withdrawn at 10 minutes intervals starting at 0900 h for 24 h. GH and PRL release were quantified with deconvolution methods. The regularity of GH and PRL release was measured with approximate entropy statistics. RESULTS: The number of GH secretory events per 24 h was higher in patients than in controls: 19 +/- 1.3 vs. 14 +/- 1.5 peaks per 24 h, respectively (P = 0.020). GH secretion rate was about one quarter lower in patients (ns), and the 24 h secretion of PRL was unchanged. Total GH production correlated negatively with the urinary excretion of free cortisol (R = 0.729, P = 0.005) and with the plasma cortisol production rate (R = 0.613; P = 0.026). The orderliness of GH and PRL secretion was appraised with the approximate entropy statistic (ApEn). For GH secretion ApEn(1,20%) in patients was 0.952 +/- 0.084 vs. 0.404 +/- 0.047 in controls, P = 1.17 x 10-4, pointing to a markedly disordered secretion in patients. Similar results were obtained for PRL secretion: patients: 1.586 +/- 0.063 vs. 1.003 +/- 0.068 in controls, P = 3.67 x 10-5. No statistically significant differences in secretory dynamics were demonstrated between the 10 patients with a microadenoma and the four with a macroadenoma. CONCLUSION: The amount of GH released spontaneously into the circulation in adult patients with pituitary-dependent Cushing's disease is inversely related to the degree of cortisol hypersecretion. However, except for severe hypercortisolism, GH secretion is relatively preserved. In addition, secretion of GH and PRL is remarkably disordered in patients with Cushing's disease. Since we could not detect differences in GH and/or PRL secretory dynamics between patients with a microadenoma and those harboring a macroadenoma, we speculate that an intrapituitary paracrine mechanism and/or elevated cortisol feedback effects may be responsible for the evident disruption of GH and PRL secretion patterns. PMID- 10792345 TI - Assessment of disease activity in Graves' ophthalmopathy by orbital ultrasonography and clinical parameters. AB - OBJECTIVE: To study if A-mode ultrasonography (US) in combination with the Clinical Activity Score (CAS) and duration of Graves' ophthalmopathy (GO) could predict the response to immunosuppression in Graves' ophthalmopathy. DESIGN: A prospective clinical study. PATIENTS AND METHODS: Fifty-six consecutive patients with moderately severe GO were treated with retrobulbar irradiation (10 fractions of 2 Gy). Before treatment the internal reflectivity in the extraocular eye muscles, the CAS, and the duration of Graves' ophthalmopathy were assessed and related to the therapeutic outcome 26 weeks after irradiation. RESULTS: Twenty eight (50%) of the patients responded favourably to radiotherapy (four patients became worse, and 24 did not change), and the eye muscle echogenecity indeed tended to be lower in the responders than in the non-responders (P = 0.09). From a receiver operator characteristics curve, a cut-off value of 30% yielded a good positive predictive value of 85%, but the negative predictive value was only 60%. The CAS (>/= 4/10) had a positive predictive value of 65%, and a negative predictive value of 56%. When combining the ultrasound with the CAS, the positive predictive value increased to 74% and the negative predictive value to 72%. Adding the duration of eye symptoms (cut-off 18 months), the prediction of response further improved: positive predictive value 79%, and negative predictive value 89%. CONCLUSIONS: A-mode ultrasonography has a rather good positive predictive value, but its poor negative predictive value precludes its use as sole activity parameter. By adding the Clinical Activity Score and duration of Graves' ophthalmopathy, the negative predictive value increased considerably. By using this combination, inactive disease can be identified more precise, permitting rehabilitative surgery at an earlier stage in these patients. PMID- 10792347 TI - Embolization in combination with radioiodine therapy for bone metastases from differentiated thyroid carcinoma. AB - BACKGROUND: The outcome for patients with bone metastases from differentiated thyroid carcinoma is worse compared to the overall prognosis of patients with differentiated thyroid carcinoma. The aim of this study is to evaluate the effect of embolization with concomitant radioiodine treatment on the serum thyroglobulin (Tg) level, pain and neurological symptoms in patients with large bone metastases from differentiated thyroid carcinoma. PATIENTS AND METHODS: Five symptomatic patients, who presented with a large unresectable bone metastasis of differentiated thyroid carcinoma were treated with radioiodine and embolization. The effect of this combined treatment was compared to the effect of radioiodine without embolization in a previously treated control group of 6 patients. Serum Tg levels, pain and neurological symptoms were scored. Both groups were treated similarly with total thyroidectomy followed by ablation with 5.55 GBq 131I and a second dose of 5.55 GBq 131I three months later, except for embolization in the embolization group, which took place between the 2 radioiodine treatments. RESULTS: In the embolization group, serum Tg at the second 131I therapy had decreased by 88.7% (median, range: 77.1-99.3%), which was significantly more compared to the decrease of serum Tg in the control group (18.6%, range: -4.7 95%, P < 0.05). CT-scanning showed a median volume reduction of the metastasis after radioiodine treatment combined with embolization of 52.5% (range: 39-80%). Both strategies resulted in a rapid relief of pain and neurological symptoms. Embolization was not accompanied with severe complications. CONCLUSIONS: This preliminary study suggests that embolization of bone metastases of differentiated thyroid carcinoma in combination with radioiodine treatment results in a significant initial reduction of serum Tg level compared to radioiodine treatment alone. This suggests a beneficial reduction in tumour burden. In this patient category, embolization appears to be a safe and well tolerated procedure. PMID- 10792346 TI - Soluble interleukin-1 receptor antagonist concentration in patients with Graves' ophthalmopathy is neither related to cigarette smoking nor predictive of subsequent response to glucocorticoids. AB - OBJECTIVE: The aim of the present study was to evaluate serum soluble interleukin 1 receptor antagonist (sIL-1RA) concentration and its relationship with the degree of cigarette smoking in patients with Graves' ophthalmopathy (GO). DESIGN AND SUBJECTS: Twenty-two consecutive GO patients (20 women, two men; age range 25 68 years, mean 48 years; 12 smokers, 10 non-smokers) submitted to IV glucocorticoid pulses over a 3-month period. MEASUREMENTS: sIL-1RA levels were measured by an immunoenzymatic assay (sensitivity, 4 ng/l; normal range, 50-290 ng/l) before glucocorticoid treatment, after two months of therapy, and 3 months after drug withdrawal. RESULTS: Thirteen patients responded to treatment (59%; five smokers and eight non-smokers), nine were non-responders (41%; seven smokers and two non-smokers). Baseline median sIL-1RA concentration did not differ in smokers and non-smokers (222 and 173 ng/l, respectively; P = 0.69). Likewise, no significant differences were found between the two groups during treatment (537 and 389 ng/l, respectively; P = 0.28); sIL-1RA concentration after treatment was higher in smokers (258 vs. 94 ng/l; P = 0.02). There was no correlation between basal sIL-1RA levels and the degree of cigarette smoking. Likewise, there was no difference in sIL-1RA levels in responders and non-responders, either at baseline (186 vs. 216 ng/l; P = 0.83), during or after treatment. CONCLUSION: Our study suggests that circulating soluble interleukin-1 receptor antagonist levels, both at baseline and during glucocorticoid treatment, are neither influenced by cigarette smoking nor predictive of subsequent response to glucocorticoid treatment. PMID- 10792348 TI - Combined pituitary hormone deficiency and pituitary hypoplasia due to a mutation of the Pit-1 gene. AB - Several mutations of the pituitary-specific transcription factor Pit-1 have been identified. We describe a girl with a mutation of the Pit-1 gene leading to a complete lack of GH, TSH and prolactin and a marked hypoplasia of the anterior pituitary gland. The patient had a homozygous nonsense-mutation at position 172 (CGA to TGA), converting arginine into a stop codon, leading to an early termination of protein translation. During the infancy period the girl had very conspicuous symptoms of hypothyroidism and the diagnosis of thyroid insufficiency preceded the diagnosis of GH-deficiency by 1.5 years. Treatment with thyroxine and GH resulted in excellent catch-up growth. PMID- 10792350 TI - The role of CD23 in allergic disease. PMID- 10792349 TI - Breast milk as an alternative source of cytokines for offspring. PMID- 10792351 TI - Airway smooth muscle function in asthma. PMID- 10792352 TI - The role of diet in the aetiology of asthma. PMID- 10792353 TI - Urban air pollution and plant-derived respiratory allergy. PMID- 10792354 TI - Defective tumour necrosis factor-alpha production in mother's milk is related to cow's milk allergy in suckling infants. AB - BACKGROUND: The precise role of leucocytes in human milk is still unresolved. OBJECTIVE: To assist in clarifying the immune mechanisms involved in the development of CMA in suckling infants, we studied the role of immunoregulatory leucocytes and their mediators in human breast milk. METHODS: The study population consisted of 43 lactating mothers and their infants, aged 0.25-8.0 months, followed-up prospectively from birth. Of these mothers, 27 had an infant with challenge-proven cow's milk allergy manifested with either skin (n = 23), gastrointestinal (n = 2) or skin and gastrointestinal symptoms (n = 3). Sixteen mothers with a healthy infant served as controls. We evaluated the spontaneous and mitogen-induced tumour necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma) production of human milk leucocytes and isolated peripheral blood lymphocytes in vitro with a commercial ELISA kit. RESULTS: TNFalpha production of breast milk leucocytes was significantly lower in the mothers with a cow's milk allergic infant, whereas IFNgamma production of these cells was comparable in the two groups. CONCLUSION: Our results suggest that in the breast milk of mothers having an infant with cow's milk allergy, the number and function of TNFalpha producing cells is defective. This might lead to a disturbance in the development of oral tolerance and thereby to the development of CMA in suckling infants. These novel results may help in clarifying the etiopathogenesis of CMA. PMID- 10792355 TI - Early BCG vaccination and reduction in atopy in Guinea-Bissau. AB - BACKGROUND: It has been proposed that certain viral and bacterial infections in early childhood may prevent allergic sensitization, by inducing Th1-type immune responses. This has led to speculation that mycobacterial vaccines might, through their Th1-stimulating properties, also protect against atopy. OBJECTIVE: To investigate whether the prevalence of atopy is lower in children who have been vaccinated with BCG in infancy than in children who have not been vaccinated. METHODS: We measured skin test reactivity to three allergens (Dermatophagoides pteronyssinus, D. farinae and cockroach) in 400 children, aged 3-14 years, as part of a follow-up study to examine the immune sequelae of measles in an urban area of Bissau, the capital of Guinea-Bissau in west Africa. Information on childhood vaccinations, including BCG in infancy, was available from child records. Of these children, 271 had been vaccinated with BCG (according to records) and 53 had not been vaccinated (no record and no BCG scar). Atopy was defined in two ways, according to the presence of any allergen reaction > or = 2 mm and any reaction > or = 3 mm. RESULTS: Of the children who had received BCG vaccine, 57 (21%) were atopic (any reaction > or = 2 mm), compared with 21 (40%) of the unvaccinated children [odds ratio, after controlling for potential confounding factors, 0.19 (95% CI 0.06-0.59)]. When atopy was defined using the 3 mm criterion, the reduction in atopy associated with BCG was greater the earlier the age at vaccination, and the largest reduction was seen in children vaccinated in the first week of life. CONCLUSION: BCG vaccination given early in infancy may prevent the development of atopy in African children. PMID- 10792356 TI - Association between HLA genes and susceptibility to toluene diisocyanate-induced asthma. AB - BACKGROUND: Only a small proportion of subjects exposed to isocyanates develop occupational asthma, suggesting individual predisposition. The human leucocyte antigen (HLA) class II molecules may play a crucial role in the development of the immune response to isocyanates. OBJECTIVES: To investigate the role of HLA class II molecules in the development of toluene diisocyanate (TDI)-induced asthma. SUBJECTS: Sixty-seven asthmatic subjects and 27 asymptomatic exposed subjects (controls) were typed at the HLA class II DQA1, DQB1 and DRB1 loci by polymerase chain reaction (PCR)-based techniques. RESULTS: The frequencies of DQA1*0104 and DQB1*0503 were significantly increased in asthmatics compared with asymptomatic exposed subjects, while DQA1*0101 and DQB1*0501 were significantly increased in asymptomatic exposed subjects. No significant difference was found in the distribution of DRB1 alleles between asthmatics and controls. CONCLUSIONS: The results of this study indicate that HLA-regulated immune mechanisms are involved in TDI-induced asthma and that, in exposed subjects, specific factors may increase or decrease the risk of developing disease. PMID- 10792357 TI - Airway inflammation and hyperresponsiveness to adenosine 5'-monophosphate in chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is often accompanied by bronchial hyperresponsiveness (BHR). Measurement of BHR may give information about airway inflammation. OBJECTIVE: To investigate the role of airway inflammation in hyperresponsiveness to adenosine 5'-monophosphate (AMP) in COPD. METHODS: We investigated inflammatory indices in induced sputum, bronchoalveolar lavage (BAL) fluid and bronchial biopsies in subjects with COPD with and without hyperresponsiveness to AMP. Twelve nonatopic subjects with COPD with hyperresponsiveness to AMP (mean +/- SD, age 63 +/- 8 years, FEV1% predicted 56 +/- 13%), six without BHR (age 60 +/- 6 years, FEV1% predicted 65 +/- 11%) and 11 healthy nonatopic controls without BHR (age 58 +/- 8 years, FEV1% predicted 104 +/- 11%) participated in the study. RESULTS: Subjects with COPD with BHR had significantly higher numbers of mucosal CD8 + and higher percentages of sputum eosinophils than those without BHR (median, 550 cells/mm2; range, 30-1340 vs 280 cells/mm2; range, 110-450, P = 0.045; and median, 2.7%; range, 0.5-8.5 vs 0.6%; range, 0-0.8 %, P = 0.0036, respectively). No differences were observed in BAL fluid. CONCLUSION: We conclude that hyperresponsiveness to AMP in COPD is associated with airway inflammation that is characterized by increased numbers of mucosal CD8 + cells and percentages of sputum eosinophils. Hyperresponsiveness to AMP may be used as a marker of airway inflammation in COPD, but its significance in the clinical course remains to be determined. PMID- 10792358 TI - Eosinophils are a feature of upper and lower airway pathology in non-atopic asthma, irrespective of the presence of rhinitis. AB - BACKGROUND: Asthma and rhinitis often co-exist and there are data to suggest that they may be two ends of the same disease spectrum. Immunohistochemical studies have shown that eosinophilia in the airways is a feature of rhinitic patients without asthma. OBJECTIVE: The aim of our study was to examine whether cellular infiltration exists in the nasal mucosa of asthmatics even in the absence of symptoms and signs of rhinitis. METHODS: Nasal mucosa biopsies were taken from 27 non-atopic subjects and comprised nine asthmatic rhinitic patients (AR), eight asthmatic non-rhinitic patients (ANR) and 10 healthy control subjects (N). Bronchial mucosa biopsies were also taken simultaneously from some of the patients (n = 10) to determine whether there was an association between cellular infiltration in the nose and the lungs. The alkaline phosphatase-anti-alkaline phosphatase (APAAP) method was used on 6 microm thick cryostat sections using monoclonal antibodies against T cells (CD4, CD8), eosinophils (EG2) and mast cells (mast cell tryptase). Slides were counted blind and results expressed as cells per field. RESULTS: The results showed that eosinophil counts were higher in both asthma groups compared with control nasal biopsies (median values AR 8.3, ANR 9.2, N 2.1 cells per field, P < 0.01). Furthermore, there was a significant correlation between eosinophil cell counts in the nose and the airways (r = 0.851 P < 0.001). No differences in eosinophil numbers were detected between the two groups of asthmatics. Also, no differences were noted for any other cell type (i.e. CD4, CD8, tryptase) among the three study groups. CONCLUSIONS: These results show that eosinophil infiltration was present in the nasal mucosa of asthmatic patients even in the absence of rhinitis, and add further support to the hypothesis that asthma and rhinitis are clinical expressions of the same disease entity. PMID- 10792359 TI - Skin prick test and serological analysis with recombinant group 2 allergens of the dust mites L. destructor and T. putrescentiae. AB - BACKGROUND: The dust mites Lepidoglyphus destructor and Tyrophagus putrescentiae are important sources of allergen in farming environments. The major allergens of the dust mites L. destructor and T. putrescentiae have been cloned and expressed as recombinant proteins. OBJECTIVE: To evaluate the use of recombinant group 2 allergens of L. destructor (rLep d 2) and T. putrescentiae (rTyr p 2) in skin prick test (SPT), and serological analysis in sensitized and non-sensitized farmers chronically exposed to dust mites. METHODS: Skin prick test with rLep d 2, rTyr p 2 and the corresponding commercial extracts was performed in 44 farmers sensitized to L. destructor and/or T. putrescentiae, and 38 control farmers. IgE and IgG subclass antibodies to the recombinant allergens were analysed by RAST and ELISA, respectively. RESULTS: Out of the 44 subjects positive in SPT to L. destructor and/or T. putrescentiae extract, 26 (59%) displayed a positive SPT to one or the other of the recombinant allergens, whereas 21 (48%) were positive to both. Significant correlations were registered between the sizes of the weals induced by rLep d 2 and rTyr p 2 and the corresponding RAST values (P < 0.001). A majority of subjects positive in SPT to the recombinant allergens had detectable IgG4 antibodies, and the levels were significantly higher in the dust mite sensitized group than in the controls (P < 0.05). No such differences were found in the IgG1 values (P > 0.05). The results obtained with rLep d 2 and rTyr p 2 correlated relatively well with each other with respect to SPT, RAST and IgG4, suggesting that the allergens have similar or shared IgE epitopes. All the control subjects had a negative SPT and RAST to rLep d 2 and rTyr p 2. CONCLUSION: Recombinant group 2 allergens from the dust mite L. destructor and T. putrescentiae represent useful tools for diagnosis of dust mite allergy. PMID- 10792361 TI - Quantitative structural and biochemical analyses of tight junction dynamics following exposure of epithelial cells to house dust mite allergen Der p 1. AB - BACKGROUND: House dust mite allergen Der p 1 is a cysteine peptidase. Previously, we have suggested that the proteolytic activity of this allergen may contribute to asthma by damaging the barrier formed by the airways epithelium. OBJECTIVE: The present study applied novel techniques to compare changes in permeability with quantitative events in tight junctions (TJs) and desmosomes (DMs) of epithelial cells exposed to Der p 1. METHODS: Confluent monolayers of Madin-Darby canine kidney (MDCK) and 16HBE14o-human bronchial epithelial cells were used as experimental models. Permeability was estimated from mannitol clearance. Digital imaging with quantification of TJs and DMs was achieved by fluorescent antibody staining and 2-photon molecular excitation microscopy (2PMEM). Biochemical changes in TJs were studied by immunoblotting, radiolabelling and immunoprecipitation. RESULTS: Der p 1 caused a time-dependent breakage of TJs and reduction in their content of the protein ZO-1. Reduction in ZO-1 immunofluorescence at TJs occurred with a small increase in the amount of diffuse, cytoplasmic immunoreactive ZO-1 staining. Morpho-logical changes in TJs occurred in synchrony with increases in epithelial permeability. DM puncta increased both in size and intensity of staining. Immunoblotting demonstrated that the disruption of TJ morphology was associated with cleavage of ZO-1 and occludin. Cells recovered from allergen exposure by de novo synthesis of occludin. CONCLUSION: Der p 1 could contribute to sensitization and allergic responses by degrading the function of the airway epithelial barrier. PMID- 10792360 TI - Biochemical and immunological characterization of a recombinant precursor form of the house dust mite allergen Der p 1 produced by Drosophila cells. AB - BACKGROUND: The major house dust mite allergen Der p 1 elicits strong IgE antibody responses in patients suffering from mite allergy. OBJECTIVE: This study reports the expression and characterization of a recombinant precursor form of Der p 1 secreted as ProDer p 1 from insect cells. METHODS: The cDNA coding for ProDer p 1 was cloned downstream to the gp67 signal peptide, starting from commercial cDNA encoding Der p 1 and PCR-amplified ProDer p 1 genomic fragment. ProDer p 1, expressed in Drosophila cells and purified from culture medium, was compared to Der p 1 isolated from mite culture, in terms of glycosylation, enzymatic activity as well as IgG- and IgE-binding capacity. RESULTS: Sequence analysis of the genomic clone of ProDer p 1 revealed that, besides two introns in the mature Der p 1 coding sequence, two introns were also present in the propeptide coding sequence. ProDer p 1 was purifed to homogeneity by a combination of ion-exchange, hydroxyapatite and gel filtration chromatographies. The precursor form of Der p 1 could be processed in vitro into mature Der p 1 under acidic and reducing conditions. Carbohydrate analysis clearly indicated that ProDer p 1 expressed from insect cells was glycosylated and that glycan structures were located only in the prosequence. ProDer p 1 displayed a similar immunoreactivity towards IgE, monoclonal and polyclonal IgG antibodies compared to natural Der p 1. Specific activity measurements using synthetic substrates clearly indicated that, contrary to natural Der p 1, ProDer p 1 was totally enzymatically inactive. CONCLUSIONS: The expression of an enzymatically inactive and highly antigenic ProDer p 1 zymogen molecule could be a suitable strategy for the development of in vitro diagnosis test as well as for specific immunotherapy. PMID- 10792362 TI - Characterization of platelet-activating factor-induced cytosolic calcium mobilization in human eosinophils. AB - BACKGROUND: Activated eosinophils play an important role in the pathogenesis of bronchial asthma and other allergic diseases, and platelet-activating factor (PAF) is a potent activator of eosinophils. OBJECTIVE: To characterize the cytosolic Ca2+ ([Ca2+]i) mobilization in human eosinophils in response to PAF. METHODS: [Ca2+]i responses to PAF were examined in human eosinophils using a microscopic fura-2 fluorescence-ratio imaging system. RESULTS: PAF caused a significant and dose-dependent increase in (Ca2+)i, which consisted of an initial rapid rise followed by a sustained elevation. This PAF-induced (Ca2+)i rise was inhibited by WEB 2086, a specific PAF receptor antagonist. The addition of 5 mM EGTA or 1 mM Ni2+ to a nominally Ca2+-free solution did not appreciably reduce the initial rise but significantly inhibited the sustained rise. The application of a protein kinase C inhibitor, Ro31-8220, augmented the sustained increase by PAF. Thapsigargin, a microsomal Ca2+ ATPase inhibitor, induced no appreciable change in a nominally Ca2+-free solution but induced a marked increase in (Ca2+)i when changed to a Ca2+-containing solution. CONCLUSIONS: The initial rapid rise and the following sustained rise in (Ca2+)i by PAF depends on Ca2+ release from the intracellular Ca2+ stores and Ca2+ influx, respectively, which are regulated by protein kinase C in human eosinophils. Furthermore, the so called Ca2+ capacitative entry is possibly involved in the Ca2+ influx from the extracellular solution in human eosinophils. PMID- 10792363 TI - Evidence to suggest that the phosphodiesterase 4 isoenzyme is present and involved in the proliferation of umbilical cord blood mononuclear cells. AB - BACKGROUND: The type 4 phosphodiesterase (PDE) isoenzyme is the main isoenzyme of PDE involved in the control of adult mononuclear cell proliferation. OBJECTIVE: To establish whether PDE isoenzymes are present in umbilical cord blood mononuclear cells by the use of selective PDE inhibitors, and to identify which PDE isoenzymes are involved in controlling the proliferation of cord blood mononuclear cells. METHODS: Cord blood was obtained from normal deliveries and mononuclear cells isolated as described previously [1] with some modifications. Mononuclear cells were then stimulated to proliferate with phytohaemagglutinin (PHA) (2 microg/mL) in the presence of selective PDE inhibitors. Proliferation was measured by [3H]-thymidine incorporation. RESULTS: The type 4 PDE inhibitors (CDP840, rolipram and RO 20-1724), and the mixed PDE3/4 inhibitor, zardaverine, produced a concentration-related inhibition of PHA-stimulated cord blood mononuclear cell proliferation (P < 0.05, ANOVA). The non-selective PDE inhibitor, theophylline, also produced a concentration-related inhibition of proliferation (P < 0.05, ANOVA). In contrast, the PDE1 inhibitor, vinpocetine, the PDE3 inhibitor, siguazodan, and the PDE5 inhibitor, zaprinast, were unable to inhibit cord blood mononuclear cell proliferation. CONCLUSION: PDE4 is present in umbilical cord mononuclear cells and is involved in the control of cord blood mononuclear cell proliferation. PMID- 10792365 TI - Inhibition of CD23 processing correlates with inhibition of IL-4-stimulated IgE production in human PBL and hu-PBL-reconstituted SCID mice. AB - BACKGROUND: CD23, the low affinity serum immunoglobulin E (IgE) receptor, is upregulated on B cells following interleukin (IL)-4 stimulation and is concomitantly cleaved to generate soluble CD23 (sCD23) fragments with cytokine like activity. OBJECTIVE: Compounds that selectively inhibit the proteolytic release of CD23 to generate sCD23 were assessed for their ability to inhibit IgE production in order to evaluate the contribution of sCD23 in the production of human IgE as well as the ability of such compounds to block IgE production. METHODS: IgE production was measured in IL-4-stimulated human peripheral blood lymphocytes (PBL) and PBL-reconstituted SCID mice in the presence of a broad spectrum matrix metalloprotease (MMP) inhibitor, a compound selective for inhibition of CD23 processing over MMPs and an anti-CD23 mAb, MHM6. RESULTS: The two compounds were equipotent in inhibiting IgE production without inhibition of IgG production by IL-4/anti-CD40-stimulated PBL. Soluble CD23 release was also shown to precede IgE accumulation in the cell-free medium. Addition of compound at later times other than day 0 in the 14 day assay resulted in progressively less inhibition of both IgE and sCD23, and exactly paralleled the effect of an anti-CD23 mAb, MHM6 on IgE levels. Both compounds also inhibited the release of CD23 from human RPMI 8866 cells adoptively transferred i. p. to mice. Doses required for inhibition of CD23 correlated well with the doses required for inhibition of IgE production in IL-4-challenged hu-PBL-SCID mice. IgE was selectively inhibited over total IgG in the SCID mice as well. CONCLUSIONS: Inhibition of CD23 processing alone is sufficient to inhibit IL-4-stimulated IgE production both in vitro and in vivo. PMID- 10792364 TI - Effect of aerosolized administration of KF19514, a phosphodiesterase 4 inhibitor, on bronchial hyperresponsiveness and airway inflammation induced by antigen inhalation in guinea-pigs. AB - BACKGROUND: Although phosphodiesterase (PDE) 3 and 4 inhibitors have received much attention for the treatment of bronchial asthma, systemic adverse effects have also been reported. OBJECTIVE: The purpose of this study was to investigate the effect of inhaled olprinone, a newly developed PDE3 inhibitor, and KF19514, a PDE1 and 4 inhibitor, on antigen-induced airway reactions in guinea-pigs. METHODS: Fifteen minutes after inhalation of olprinone (0.1 or 1.0 mg/mL) and KF19514 (0.1 or 0.01 mg/mL), animals were given an antigen challenge. Bronchial hyper-responsiveness and bronchoalveolar lavage fluid cell analysis were performed 24 h after the antigen challenge. RESULTS: Inhalation of olprinone and KF19514 caused a dose-related inhibition of antigen-induced bronchoconstriction. Antigen inhalation significantly increased bronchoconstrictor responses to methacholine, and airway accumulation of neutrophils and eosinophils, 24 h after the antigen challenge. These responses were dose-dependently prevented by KF19514, but not by olprinone. CONCLUSION: The results indicate that inhaled PDE inhibitors might be useful for treatment of bronchial asthma. PMID- 10792366 TI - The role of CD23 on allergen-induced IgE levels, pulmonary eosinophilia and bronchial hyperresponsiveness in mice. AB - BACKGROUND: The role of Immunoglobulin (Ig)E in inflammation is the subject of considerable study and a number of studies have shown conflicting evidence for its role in eosinophil recruitment and bronchial hyperresponsiveness in a number of murine models. The low affinity IgE receptor, CD23, is known to act as a negative regulator of IgE production and we have used knockout mice deficient in CD23 to investigate the role of IgE in eosinophil recruitment and bronchial hyperresponsiveness in a murine model of airway inflammation. OBJECTIVE: To study the role of the low affinity FcepsilonII receptor, CD23 in IgE production, lung inflammation and bronchial hyperresponsiveness. METHODS: Wild-type and CD23 knockout C57Bl/6 mice (CD23-/-) were immunized by intraperitoneal injection with ovalbumin on days 0 and 14 and challenged with aerosolized antigen on day 21 for a period of up to 1 week. Blood samples, bronchoalveolar lavage and lung tissue samples were obtained to determine serum IgE levels and inflammatory cell numbers, respectively. Furthermore, airway resistance was measured to increasing concentrations of aerosolized 5-hydroxytryptamine in order to evaluate the effect of CD23 deficiency on bronchial hyperresponsiveness to antigen challenge. RESULTS: Sensitization of wild-type C57Bl/6 mice to ovalbumin resulted in elevated levels of total serum IgE and ovalbumin-specific IgE, which was significantly augmented in CD23 knockout C57Bl/6 mice (CD23-/-). A significant increase in the percentage of eosinophils recovered in bronchoalveolar lavage fluid from wild-type and CD23-/- mice was observed 24 h following 3 or 7 days aerosol exposure with ovalbumin (10 mg/mL). At 3 days, the increase in the percentage of eosinophils was significantly greater in CD23-/- groups. Immunohistochemical analysis of lungs sections revealed the presence of CD3+, CD4+ and CD23+ cells in wild-type mice but a lack of immunofluorescence of CD23+ cells in CD23-/- mice. In wild-type ovalbumin-immunized mice, bronchial hyperresponsiveness to aerosolized 5-hydroxytryptamine was observed following a 3 day antigen challenge, which was significantly greater in CD23-/- ovalbumin immunized mice. CONCLUSION: These studies demonstrate that CD23-/- mice have increased capacity to produce IgE consistent with the view of a negative feedback role for membrane-bound CD23 and under such conditions, may account for the greater numbers of eosinophils recruited to the airways and bronchial hyperresponsiveness observed following acute but not chronic antigen challenge. PMID- 10792367 TI - Why do some dietary migraine patients claim they get headaches from placebos? AB - BACKGROUND: In six double-blind studies involving 182 tests of dietary migraine patients sensitive to tyramine and beta-phenylethylamine, 18% reported headaches from placebos which were all concealed in gelatin capsules. OBJECTIVE: The purpose of this research was to test a hypothesis: gelatin is partially hydrolysed animal protein; (partially) hydrolysed vegetable protein (PHVP) is known to cause migraine; perhaps the gelatin caused some of the headaches. METHOD: The author tested this hypothesis on himself because he suffers from dietary migraine. He proved this in a double-blind test with tyramine hydrochloride (TYH). The amount required for the test was so small (1 mg) that it was tasteless and capsules were unnecessary. The author then undertook tests with a capsule, PHVP, monosodium glutamate (MSG) aspartame (a dipeptide) and TYH, adjusting quantities to give a moderate headache. Samples were mixed with foods to simulate normal eating: the capsule with potato chips, aspartame with orange juice and the rest with cottage cheese or ricotta cheese. Times were measured from ingestion (1) to start of the headache and (2) to maximum headache intensity. Each experiment was repeated three times. The headaches were relieved with caffeine. RESULTS: Of eight double-blind test samples, the author identified correctly the two placebos and five of the six samples containing tyramine. Quantities giving moderate headaches were: 1 gelatin capsule, 400 mg MSG, 118 mg PHVP, 4.0 mg aspartame and 1.0 mg TYH. Typical times for the three repetitions of the two time periods were 8, 9 and 11 and 17, 19 and 22 min. CONCLUSIONS: Capsules may give headaches to dietary migraine patients that are similar to those from foods. This would explain some of the headaches of patients from placebos. The double-blind test and the repeatability of the time measurements demonstrated the validity of the experiments. PMID- 10792368 TI - Selective IgA deficiency (SIgAD) and common variable immunodeficiency (CVID). PMID- 10792369 TI - Effective vaccination against tuberculosis-a new ray of hope. PMID- 10792370 TI - Cytolytic mechanisms of intraepithelial lymphocytes in coeliac disease (CoD). AB - The effector arm of the mucosal immune system comprises lymphocytes scattered at intraepithelial and lamina propria levels. Intraepithelial lymphocytes (IEL) are a large population of oligoclonal resting cells which exhibit phenotypic and functional characteristics of cytolytic T cells when activated. Several mechanisms have been demonstrated to account for their cytotoxicity. Among them, one is mediated by perforin and granzyme molecules, another is mediated by Fas ligand (FasL) which delivers apoptotic signals through Fas receptor on target cells. There is good evidence that a flat intestinal mucosa may be produced by activated T cells. The aim of our study was to evaluate FasL and perforin expression by IEL, and its possible correlation with the increased enterocyte apoptosis in coeliac mucosa. Endoscopic duodenal biopsy specimens from 10 untreated coeliac patients, 10 treated coeliac patients, and 10 biopsied controls were evaluated for enterocyte apoptosis by terminal deoxynucleotidyl transferase mediated digoxigenin-deoxyuridine triphosphate nick end label method, for perforin expression by immunohistochemistry, and for FasL expression by immunocytochemistry. In untreated CoD there was a significant increase of percentage of both FasL+ and perforin+ IEL which positively correlated with enterocyte apoptosis in comparison with controls. All these parameters were significantly lower in treated CoD, even though they did not normalize. Our study demonstrates that in untreated CoD FasL and perforin expression by IEL is increased, and significantly correlates with the level of enterocyte apoptosis. PMID- 10792371 TI - Increased production of matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 by inflamed mucosa in inflammatory bowel disease. AB - Inflammatory bowel diseases (IBD) are characterized by a sustained inflammatory cascade that gives rise to the release of mediators capable of degrading and modifying bowel wall structure. Our aims were (i) to measure the production of matrix metalloproteinase-3 (MMP-3), and its tissue inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), by inflamed and uninflamed colonic mucosa in IBD, and (ii) to correlate their production with that of proinflammatory cytokines and the anti-inflammatory cytokine, IL-10. Thirty-eight patients with IBD, including 25 with Crohn's disease and 13 with ulcerative colitis, were included. Ten controls were also studied. Biopsies were taken from inflamed and uninflamed regions and inflammation was graded both macroscopically and histologically. Organ cultures were performed for 18 h. Tumour necrosis factor-alpha (TNF-alpha), IL-6, IL-1beta, IL-10, MMP-3 and TIMP-1 concentrations were measured using specific immunoassays. The production of both MMP-3 and the TIMP-1 were either undetectable or below the sensitivity of our immunoassay in the vast majority of uninflamed samples either from controls or from those with Crohn's disease or ulcerative colitis. In inflamed mucosa, the production of these mediators increased significantly both in Crohn's disease (P < 0.01 and 0.001, respectively) and ulcerative colitis (P < 0.001 and 0.001, respectively). Mediator production in both cases was significantly correlated with the production of proinflammatory cytokines and IL-10, as well as with the degree of macroscopic and microscopic inflammation. Inflamed mucosa of both Crohn's disease and ulcerative colitis show increased production of both MMP-3 and its tissue inhibitor, which correlates very well with production of IL-1beta, IL-6, TNF alpha and IL-10. PMID- 10792372 TI - The antagonism of interferon-gamma (IFN-gamma) by heparin: examination of the blockade of class II MHC antigen and heat shock protein-70 expression. AB - IFN-gamma is a pleiotropic cytokine that is primarily involved in the regulation of immune cell activation and the development of tissue inflammation. It is capable of activating a range of non-immune cells, including those of the vascular endothelium. These cells respond by increasing the expression of intracellular and cell-surface molecules such as class II MHC antigens and adhesion molecules that, together, increase the tendency for interaction with immune cells. It is known that IFN-gamma can bind cell surface and extracellular heparan sulphate. Furthermore, soluble heparin can inhibit the function of this cytokine, presumably by competitive displacement from the cell surface, resulting in the failure of normal receptor signal transduction. In this study it is shown that heparin can prevent normal induction of the class II transactivator and heat shock cognate protein-70 in an IFN-gamma-treated endothelial cell line. Both of these molecules are dependent on the activation of intracytoplasmic STAT-1, which is the most receptor proximal component of their respective induction pathways. This provides further evidence for the blockade by heparin of ligand activation of the specific IFN-gamma receptor. PMID- 10792373 TI - Cryofibrinogenaemia: a study of 49 patients. AB - The purpose of this study was to characterize the clinical features and components of 30 patients with isolated cryofibrinogen (CF) versus those of 19 patients with combined CF and cryoglobulins (CG). Secondary forms of cryofibrinogenaemia associated with collagen disorders, infectious or malignant diseases, were significantly more frequent in patients with combined CF and CG than those with isolated CF (79 versus 47%, P = 0.02). Both groups of CF patients presented predominantly cutaneous symptoms (77% in isolated CF; 58% in combined CF + CG), and less frequently venous and/or arterial thrombosis (13% in isolated CF; 3% in combined CF + CG). Patients with idiopathic forms of CF, and particularly those without CG, suffered essentially from recurrent painful skin ulcers, mainly triggered by cold exposure. Patients with isolated CF had higher mean plasma concentrations of CF than those with combined CF + CG (1. 61 +/- 1.26 versus 0.82 +/- 1.18 g/l, respectively; P = 0.004), but there was no correlation between the CF plasma level and either the severity of symptoms or the sensitivity to cold. In patients with isolated CF, fibronectin was suggested (by precipitation analysis) to be a major component of the cryoprecipitate, whereas immunoglobulins were rarely present (in only three out of 30 patients). By contrast, in the majority of patients (78%) with combined CF and CG, the CF consisted mainly of immunoglobulins of the same class as those characterizing the associated CG. Analysis of the CG precipitate revealed the presence of fibronectin but not fibrinogen, alpha1-antitrypsin and alpha2-macroglobulin. In conclusion, isolated and combined cryofibrinogenaemia are associated with different clinical signs requiring different clinical management, but there is no evidence as yet for a causal role of the cryoprecipitates in the differences observed. PMID- 10792374 TI - Proteolytic components of serum IgG preparations. AB - Chemical catalysis, an effector mechanism utilized by fully assembled antibodies, can also be mediated by the isolated antibody subunits. Because trace amounts of free light chains (L chains) are present in IgG preparations, a detailed study was undertaken to identify the constituents responsible for the polyreactive proteolytic activity of IgG purified from human sera, determined as the extent of cleavage of the model peptide substrate Pro-Phe-Arg-methylcoumarinamide. Two proteolytic species with approximate mass of 50 kD and 150 kD were separated by repetitive gel filtration in a denaturing solvent (6 M guanidine hydrochloride). The activity of the renatured 50-kD fraction (in fluorescence units/microg protein) was more than 45-fold greater than of the 150-kD fraction. Both fractions lost the activity following immunoadsorption on immobilized anti-IgG antibody. Fab fragments prepared from the 150-kD IgG fraction retained the activity. Reducing and non-reducing SDS-electrophoresis suggested the 50-kD fraction isolated from the IgG preparations to be a mixture of heavy chain (H chain) monomers and disulphide bonded L chain dimers. Electrophoretically homogeneous monomers of 50-kD H chains and 25-kD L chains were prepared by gel filtration of reduced and alkylated IgG from seven human subjects. Each of the alkylated L chain preparations displayed the proteolytic activity. The activity in alkylated H chains was undetectable or only marginally greater than the background values. L chain dimers appear to be the major species responsible for the polyreactive proteolytic activity of serum IgG preparations, with a smaller contribution furnished by tetrameric IgG. PMID- 10792375 TI - Altered Th1/Th2 commitment in human CD4+ T cells with ageing. AB - The human immune system undergoes continuous remodelling with the advancement of age. Since age-associated functional alterations in the immune system could be caused by a possible change in helper T cell regulation in elderly subjects, we comparatively studied the function of CD4+ T cells in peripheral blood obtained from both young and old healthy volunteers. Upon cell activation by phorbol myristate acetate and ionomycin, the proportion of CD4+ T cells containing interferon-gamma (IFN-gamma) was found to be greater in the old subjects. Utilizing a co-culture system, which activated CD4+ T cells via the TCR/CD3 complex and CD28, we found that CD4+ T cells from the old subjects secreted more IFN-gamma and IL-2, but less IL-4, than those from the young subjects. Upon cell activation by co-culture, CD4+ T cells from the old subjects expressed more CD26, CD40L, and LFA-1, but less CD30, than those from the young. These results together suggest that the microenvironment in which CD4+ T cells develop in older people may cause production of more cells committed to Th1 than that in younger subjects. PMID- 10792376 TI - The 19-kD antigen and protective immunity in a murine model of tuberculosis. AB - The 19-kD antigen is a cell wall-associated lipoprotein present in Mycobacterium tuberculosis and in bacille Calmette-Guerin (BCG) vaccine strains. Expression of the 19-kD antigen as a recombinant protein in two saprophytic mycobacteria-M. vaccae and M. smegmatis-resulted in abrogation of their ability to confer protection against M. tuberculosis in a murine challenge model, and in their ability to prime a DTH response to cross-reactive mycobacterial antigens. Induction of an immune response to the 19-kD antigen by an alternative approach of DNA vaccination had no effect on subsequent M. tuberculosis challenge. These results are consistent with a model in which the presence of the 19-kD protein has a detrimental effect on the efficacy of vaccination with live mycobacteria. Targeted inactivation of genes encoding selected antigens represents a potential route towards development of improved vaccine candidates. PMID- 10792377 TI - The proportion of circulating gammadelta T cells increases after the first week of onset of tularaemia and remains elevated for more than a year. AB - In various human intracellular bacterial diseases, an increase of the proportion of circulating Vgamma9Vdelta2 T cells has been observed. The prevalence of the finding among infected subjects and the time course of the elevation remain to be investigated. In the present study, comprising blood samples from a large number of cases of ulceroglandular tularaemia, the percentage of Vgamma9Vdelta2 T cells within the first week of onset of disease (5.3 +/- 0.7% (mean +/- s.e.m.)) did not differ from that of control subjects (5.3 +/- 0. 8%). Thereafter, percentages increased rapidly and within the interval of 8-40 days mean levels were > 20% (P < 0.001). Of 45 individuals sampled within 3 months of onset, 42 showed a percentage of Vgamma9Vdelta2 T cells of > 10%. Significantly increased levels were still recorded at 18 months (13.8 +/- 2.4%; P < 0.05) but not at 24 months (10.2 +/- 2.1%; P > 0.10). Thus, a consistent increase of circulating Vgamma9Vdelta2 T cells was demonstrated in tularaemia. The initial delay and the prolonged course of elevation may suggest a role in immunoregulation and/or immunological memory. Furthermore, the percentage of gammadelta T cells expressing tumour necrosis factor-alpha in response to phorbol myristate acetate was decreased during the first week and up to 40 days after onset, possibly reflecting the modulation of an inflammatory response. PMID- 10792378 TI - Elevated humoral immune response to heat shock protein 60 (hsp60) family in periodontitis patients. AB - The presence of antibodies to the 60-kD human and Porphyromonas gingivalis GroEL hsp60 in the sera and inflamed gingival tissues of periodontitis patients was examined. In order to obtain the antigens, recombinant plasmids carrying human hsp60 and P. gingivalis GroEL genes were constructed and expressed as histidine tagged recombinant proteins. Immunoreactivities of these proteins were confirmed by MoAbs specific to mammalian hsp60 and cross-reactive with both mammalian and bacterial hsp60. Western blot analysis clearly demonstrated that the number of periodontitis patients showing a positive response to P. gingivalis GroEL was higher than the number of periodontally healthy subjects. Furthermore, anti-P. gingivalis GroEL antibody was detected in all samples of gingival tissue extracts. For human hsp60, a higher frequency of seropositivity was found in the periodontitis patients than in the healthy subjects. In addition, the periodontitis patients demonstrated stronger reactivity compared with the healthy subjects. Quantitative analysis of serum antibodies by ELISA also demonstrated that the levels of antibodies in the sera of patients were significantly higher than those of control subjects. In the gingival tissue extracts, seven out of 10 patients demonstrated a positive response to human hsp60 and tso of these demonstrated strong positivity. Affinity-purified serum antibodies to human hsp60 and P. gingivalis GroEL from selected patients reacted with P. gingivalis GroEL and human hsp60, respectively, suggesting cross-reactivity of antibodies. These results suggest that molecular mimicry between GroEL of the periodontopathic bacterium P. gingivalis and autologous human hsp60 may play some role in immune mechanisms in periodontitis. PMID- 10792380 TI - Specificity of anti-phospholipid antibodies in infectious mononucleosis: a role for anti-cofactor protein antibodies. AB - The antigen specificity of anti-phospholipid antibodies in infectious mononucleosis (IM) was studied using ELISA for the detection of anti-beta2 glycoprotein I (beta2-GPI), anti-annexin V, anti-protein S and anti-prothrombin antibodies and TLC immunostaining for the detection of anti-phospholipid antibodies. This technique enabled us to look at antibodies reacting to 'pure' phospholipid antigens in the absence of protein contamination. Sera from 46 patients with IM, 18 with systemic lupus erythematosus (SLE), 21 with primary anti-phospholipid antibody syndrome (PAPS), 50 with Helicobacter pylori infection and 30 healthy blood donors were tested. This study highlights anti-phospholipid antibodies in patients with IM as specific 'pure' anti-cardiolipin antibodies, while in PAPS and SLE patients anti-phosphatidylserine and anti phosphatidylethanolamine antibodies were also found. This investigation also shows that the anti-cardiolipin antibodies found in IM can be present with anti cofactor protein antibodies. The higher prevalence of anti-cofactor antibodies found in IM sera than in Helicobacter pylori sera may be due to the immunostimulatory effect and/or the polyclonal activation often observed in course of Epstein-Barr virus infection. However, anti-beta2-GPI and, to a lesser extent, anti-prothrombin antibodies occur with a significantly lower prevalence in IM than in PAPS patients. This finding suggests that these antibodies should be regarded as the expression of the broad autoimmune syndrome involving the phospholipid-binding plasma proteins. PMID- 10792379 TI - Characterization of the expanded T cell population in infectious mononucleosis: apoptosis, expression of apoptosis-related genes, and Epstein-Barr virus (EBV) status. AB - Infectious mononucleosis (IM), a manifestation of primary infection with EBV, is characterized by a massive expansion of the T cell population. In this study we examined this expanded T cell population regarding its EBV status, its proliferative and apoptotic activity, and its expression of apoptosis-related genes. Whereas previous studies were performed on ex vivo cultures or on peripheral blood, our investigations included in vivo analysis of IM tonsillectomy specimens (14 cases) by in situ hybridization for viral RNA (EBERs) combined with immunohistochemistry (IHC; CD3, CD45RO, CD20, CD79a, Ki-67, Bcl-2, Bax, Fas, FasL) and the TUNEL method. Of the EBER+ cells 50-70% showed expression of the B cell markers CD20/CD79a. The remainder of the EBER+ cells expressed neither B nor T cell antigens. No co-expression of EBERs and T cell antigens was detected in any of the specimens. In accordance with a high rate of apoptosis (up to 2.37%) within the expanded T cell population, Bcl-2 expression was drastically reduced and FasL expression remarkably increased. The levels of Bax and Fas expression showed no or moderate up-regulation. In conclusion, the massive expansion of IM T cells is not caused by EBV infection of these cells but merely represents an intense immune reaction. Through altered expression of Bcl-2/Bax and Fas/FasL, the activated T cells are subject to enhanced apoptosis while residing within the lymphoid tissue, which eventually allows the efficient silencing of this potentially damaging T cell response. PMID- 10792381 TI - Frequent reversible membrane damage in peripheral blood B cells in human T cell lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP). AB - Apoptosis in peripheral blood lymphocyte populations in HTLV-I-infected people in vivo was examined, to study the lymphocyte dynamics in HTLV-I infection. Freshly isolated lymphocytes from 10 non-infected healthy people, eight asymptomatic HTLV I carriers and 15 patients with HAM/TSP were stained with FITC-labelled annexin V to detect phosphatidylserine (PS) residue exposure at the outer plasma membrane leaflet as an early marker of apoptosis. There was no significant difference in annexin V positivity in CD4+ and CD8+ lymphocytes between non-infected subjects, asymptomatic carriers and HAM/TSP patients, but there was a greatly increased exposure of PS on CD19+ lymphocytes (B cells) detected by FITC-annexin V in 12 out of 15 (80%) HAM/TSP patients, while only two out of eight (25%) asymptomatic carriers and none of the non-infected healthy people showed this aberrant PS exposure on B cells. The intensity of annexin V staining of B cells in HAM/TSP was intermediate, as distinct from the high annexin V staining on advanced apoptotic cells. However, annexin V positivity was decreased when the cells were stained after 24 h of culture, suggesting that the intermediate PS exposure on the B cell in HAM/TSP is not a consequence of an apoptotic process, but rather reflects reversible membrane damage. B cells with PS exposure in vivo might provide a site for coagulation and inflammation, and so contribute to the pathogenesis of HAM/TSP and its complications. PMID- 10792384 TI - Chemokines and soluble adhesion molecules in renal transplant recipients with cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is associated with leucocyte infiltration in various organs, which supports a role for chemokines and adhesion molecules in the pathogenesis of CMV infection. In a prospectively conducted study of renal transplant recipients, 10 patients with CMV disease, five patients with asymptomatic CMV infection and 10 patients who did not have any CMV infection were included. During CMV infection, and in particular during CMV disease, plasma levels of the chemokines IL-8, macrophage inflammatory protein-1alpha (MIP 1alpha) and monocyte chemotactic protein-1 (MCP-1) and the soluble adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and L-selectin increased and were positively correlated with the degree of CMV pp65 antigenaemia. Furthermore, a decrease in plasma levels of these chemokines and adhesion molecules was observed following ganciclovir therapy in the patients with CMV disease. This could suggest a role for these molecules in the pathogenesis of CMV infection. PMID- 10792382 TI - Hydroxyurea inhibits the transactivation of the HIV-long-terminal repeat (LTR) promoter. AB - HIV-1 gene expression is regulated by the promoter/enhancer located within the U3 region of the proviral 5' LTR that contains multiple potential cis-acting regulatory sites. Here we describe that the inhibitor of the cellular ribonucleoside reductase, hydroxyurea (HU), inhibited phorbol myristate acetate- or tumour necrosis factor-alpha-induced HIV-1-LTR transactivation in both lymphoid and non-lymphoid cells in a dose-dependent manner within the first 6 h of treatment, with a 50% inhibitory concentration of 0.5 mM. This inhibition was found to be specific for the HIV-1-LTR since transactivation of either an AP-1 dependent promoter or the CD69 gene promoter was not affected by the presence of HU. Moreover, gel-shift assays in 5.1 cells showed that HU prevented the binding of the NF-kappaB to the kappaB sites located in the HIV-1-LTR region, but it did not affect the binding of both the AP-1 and the Sp-1 transcription factors. By Western blots and cell cycle analyses we detected that HU induced a rapid dephosphorylation of the pRB, the product of the retinoblastoma tumour suppressor gene, and the cell cycle arrest was evident after 24 h of treatment. Thus, HU inhibits HIV-1 promoter activity by a novel pathway that implies an inhibition of the NF-kappaB binding to the LTR promoter. The present study suggests that HU may be useful as a potential therapeutic approach for inhibition of HIV-1 replication through different pathways. PMID- 10792383 TI - Decreased CD95 expression on naive T cells from HIV-infected persons undergoing highly active anti-retroviral therapy (HAART) and the influence of IL-2 low dose administration. Irhan Study Group. AB - The functional recovery of the immune system in HIV-infected persons receiving HAART and the role of adjuvant immune therapy are still matters of intensive investigation. We analysed the effects of HAART combined with cytokines in 22 naive asymptomatic individuals, randomized to receive HAART (n = 6), HAART plus a low dose (1000 000 U/daily) of rIL-2 (n = 8), and HAART plus rIL-2 after previous administration of granulocyte colony-stimulating factor (n = 8). After 3 months of therapy, increased CD4+ T cell counts and diminished viral loads were observed in all patients, independently of cytokine addition. A decreased expression of CD95 (Apo 1/Fas) was evident in all groups when compared with values before therapy. The percentages of peripheral blood mononuclear cells (PBMC) expressing CD95 after therapy decreased by 15%, 22% and 18% in the three treatment groups, respectively (P < 0.05). Analysis of PBMC subsets demonstrated that CD95 expression was significantly reduced on CD45RA+CD62L+ naive T cells (25.3%, 22.4%, and 18.6%, respectively; P < 0.05) in each group, after therapy. Accordingly, all patients showed a reduced rate of in vitro spontaneous apoptosis (P < 0.05). Another effect induced by HAART was a significant increase in IL 2Ralpha expression on total PBMC (P < 0.05), independently of cytokine addition. Altogether, our results suggest that very low dose administration of rIL-2 (1000 000 U/daily) may be not enough to induce a significant improvement in the immune system as regards HAART alone. The employment of higher doses of recombinant cytokines and/or different administration protocols in clinical trials might however contribute to ameliorate the immune reconstitution in patients undergoing HAART. PMID- 10792385 TI - The role of Fcgamma receptor polymorphisms and C3 in the immune defence against Neisseria meningitidis in complement-deficient individuals. AB - Individuals with either a late (C5-9) complement component deficiency (LCCD) or properdin deficiency are at increased risk to develop meningococcal disease, often due to serogroups W135 and Y. Anti-meningococcal defence in both LCCD persons and properdin-deficient individuals without bactericidal antibodies depends mainly on phagocytosis. Three types of opsonin receptors are involved in phagocytosis by polymorphonuclear cells (PMN). These represent the polymorphic FcgammaRIIa (CD32) and FcgammaRIIIb (CD16b) receptors, and the C3 receptor CR3 (CD11b/CD18). When the distribution of FcgammaRIIa and FcgammaRIIIb allotypes was assessed in 15 LCCD and in 15 properdin-deficient patients with/without previous meningococcal disease, we found the combination of FcgammaRIIa-R/R131 with FcgammaRIIIb-NA2/NA2 allotypes to be associated with previous meningococcal disease (odds ratio 13.9, Fisher's test P = 0.036). No such relation was observed in the properdin-deficient patients. The importance of FcgammaRIIa allotypes was also demonstrated using in vitro phagocytosis assays. PMN from FcgammaRIIa-R/R131 homozygous donors internalized IgG2 opsonized meningococci W135 significantly (P < 0.05) less than PMN from FcgammaRIIa-H/H131 donors. When properdin-deficient serum was tested, it was observed that reconstitution with properdin resulted in enhanced PMN phagocytosis of the W135 meningococci (P = 0.001). This enhanced phagocytosis was parallelled by an increase in C3 deposition onto the opsonized meningococci W135 (r = 0.6568, P = 0. 01). We conclude that the occurrence of meningococcal disease in LCCD patients is associated with certain FcgammaR allotypes. Properdin-deficient individuals are susceptible to meningococcal disease because of an insufficient C3 deposition on the surface of meningococci, resulting in insufficient phagocytosis. PMID- 10792386 TI - Kinase mutant Btk results in atypical X-linked agammaglobulinaemia phenotype. AB - X-linked agammaglobulinaemia (XLA) is a B cell humoral abnormality arising from mutations in the gene encoding Bruton's tyrosine kinase (Btk). The phenotype of XLA can be variable, with some individuals having a less severe immunophenotype, although in most cases this cannot be correlated with the Btk mutation or expression of Btk protein. In this study we describe clinical and immunological heterogeneity within the same pedigree. Analysis of the genetic defect identified a missense mutation in the kinase domain of Btk which, unusually, preserved Btk protein expression but at reduced levels, and also considerably diminished autophosphorylation activity. Structural analysis of the effect of this mutation on the kinase domain suggests that this mutation is not an integral part of the ATP or substrate binding domains but may affect the interaction of the kinase domain with its own kinase domain and other substrates. Together, these data may provide an explanation for the variable XLA phenotype. PMID- 10792387 TI - Follow up of patients with chronic granulomatous disease diagnosed since 1990. AB - Outcomes for children with chronic granulomatous disease (CGD) have historically been poor, but significant improvements have occurred with the use of effective antibacterial prophylaxis. The present study aimed to document the clinical course of a cohort of children diagnosed with CGD since 1990 in a single centre. Twenty-one patients were identified, with a median age at last assessment of 4 years and 5 months. A third of these children were diagnosed in infancy because of a positive family history. Of the remaining, there was a median delay between the onset of symptoms and diagnosis of 13 months. No invasive or fungal infections were documented after diagnosis, nor were there any deaths in this cohort. A variety of non-infectious complications were noted, which responded well to steroids. As a group, these children were thriving and weight and height distributions fell within the population norm. All patients were receiving antibacterial prophylaxis, 90% with co-trimoxazole, and all but one patient were receiving a prophylactic anti-fungal agent (itraconazole). Both drugs were well tolerated. In conclusion, this cohort of patients, diagnosed in the last decade, tolerated antibacterial and anti-fungal prophylaxis well and on this regimen have a significantly decreased incidence of infection when compared with historical cohorts. Careful follow up of patients who have had aggressive antibacterial and anti-fungal prophylaxis should continue. The data reported on this cohort of patients should inform the debate about the use of more aggressive treatments, such as bone marrow transplantation, in this disease. PMID- 10792388 TI - Diesel exhaust particles up-regulate expression of intercellular adhesion molecule-1 (ICAM-1) in human bronchial epithelial cells. AB - Epidemiological and experimental studies suggest that diesel exhaust particles (DEP) may play an active role in the increased respiratory mortality and morbidity. We have shown that DEP augmented the production of inflammatory cytokines by human airway epithelial cells in vitro. ICAM-1 has been shown to play an important role in the local accumulation of inflammatory cells. We studied the effect of DEP on ICAM-1 gene expression and surface expression in human bronchial epithelial cell line BEAS-2B. DEP (5-50 microg/ml) showed a stimulatory effect on ICAM-1 mRNA levels as evaluated by reverse transcription polymerase chain reaction (RT-PCR). Flow cytometric analysis demonstrated an increased ICAM-1 expression on the epithelial cell surfaces. The soluble form of ICAM-1 molecules was also increased by the stimulation of DEP. In vitro neutrophil attachment onto DEP-stimulated epithelial cells was augmented, which was partially blocked by anti-ICAM-1 neutralizing antibody. Finally, these events were significantly inhibited by pretreatment with anti-oxidants pyrrolidine dithiocarbamate and N-acetyl cysteine, and p38 mitogen activated protein kinase (MAPK) inhibitor SB203580. These findings suggested that DEP induced up regulation of ICAM-1 gene, and this process might be largely dependent on oxidant mediated NF-kappaB activation and p38-MAPK pathways. PMID- 10792389 TI - Treatment of passively transferred experimental autoimmune myasthenia gravis using papain. AB - Antibody-mediated acetylcholine receptor (AChR) loss at the neuromuscular junction, the main cause of the symptoms of myasthenia gravis, is induced by bivalent or multivalent antibodies. Passive transfer of experimental autoimmune myasthenia gravis (EAMG) can be induced very efficiently in rats by administration of intact MoAbs directed against the main immunogenic region (MIR) of the AChR, but not by their monovalent Fab fragments. We tested whether papain, which has been used therapeutically in autoimmune and other diseases, is capable of preventing EAMG by in vivo cleavage of the circulating anti-AChR antibodies into Fab fragments. EAMG was induced in 4-week-old female Lewis rats by i.p. injection of anti-MIR mAb35. A total of 0.75 mg of papain was given as one or three injections 3-7 h after MoAb injection. The mAb35 + papain-treated animals developed mild weakness during the first 30 h and subsequently recovered, while all animals that received only mAb35 developed severe myasthenic symptoms and died within 24-30 h. Animals treated only with papain showed no apparent side effects for up to 2 months. Serum anti-AChR levels in mAb35 + papain-treated rats decreased within a few hours, whereas in non-papain-treated rats they remained high for at least 30 h. Muscle AChR in mAb35 + papain-treated animals was partially protected from antibody-mediated degradation. These results show that treatment of rats with papain can prevent passively transferred EAMG without any apparent harm to the animals, and suggest a potential therapeutic use for proteolytic enzymes in myasthenia gravis. PMID- 10792390 TI - Immunosuppressant effect of gold on IgG subclasses and IgE; evidence for sparing of Th2 responses. AB - We set out to examine the effect of gold treatment on the Th2-dependent antibodies IgG4 and IgE in relation to other IgG subclasses in patients with rheumatoid arthritis (RA). Eighty-five gold-treated RA patients and 82 RA controls were studied. Serum IgG subclass concentrations were measured by ELISA, IgE was measured by automated enzyme immunoassay. Samples were studied serially in 13 gold-treated patients and in 11 patients with gold-induced adverse events. There was a significant reduction in the concentration of IgG1, IgG2 and IgG3 in gold-treated RA patients compared with RA controls (P 0.004-0.019), whereas IgG4 was less significantly reduced in gold-treated patients (P = 0.044) and there was no difference in IgE. In serial samples there was a significant fall in the concentration of IgG1 (P = 0.001), IgG2 (P = 0.001) and IgG3 (P = 0.026) with time but no change in IgG4 and IgE. The development of gold-induced adverse events was not associated with any change in the concentration of each IgG subclass or IgE. Deficiencies of IgG subclasses were found in 30% of gold-treated RA patients and 8.5% of RA controls, and were associated in gold-treated patients with a longer disease duration (P = 0.003) and with erosive disease (P = 0. 03). IgG2 was affected most frequently and in the majority of these cases subnormal specific IgG2 binding to widespread polysaccharide antigens (Pneumovax II) was found. Gold induces an overall immunosuppressant effect on IgG subclasses, with a deficiency in 21. 5%, adjusted for controls. The effect on the Th2-dependent antibodies IgG4 and IgE is less marked, suggesting a sparing of Th2 responses. PMID- 10792392 TI - Fine specificity of autoantibodies to calreticulin: epitope mapping and characterization. AB - Extracellular calreticulin (CRT) as well as anti-CRT antibodies have been reported in patients with various autoimmune disorders and CRT has been implicated in 'epitope spreading' to other autoantigens such as the Ro/SS-A complex. In addition, antibodies against parasite forms of the endoplasmic reticulum chaperone, CRT, have been found in patients suffering from onchocerciasis and schistosomiasis. In this study, we screened sera for anti-CRT antibodies from patients with active and inactive systemic lupus ertythematosus (SLE) and primary or secondary Sjogren's syndrome. Approximately 40% of all SLE patients were positive for anti-CRT antibodies. The antigenic regions of CRT were determined using full length CRT and fragments of CRT prepared in yeast and Escherichia coli, respectively. Synthetic 15mer peptides corresponding to the major autoantigenic region of CRT (amino acids 1-289), each one overlapping by 12 amino acids, were used to map the B cell epitopes on the CRT protein recognized by autoimmune sera. Major antigenic epitopes were found to be associated with the N-terminal half of the protein in 69% of the SLE sera from active disease patients, while the C-domain was not antigenic. Major epitopes were found to be reactive with antibodies in sera from SLE patients with both active and inactive disease, spanning different regions of the N and P-domains. Sera from both healthy and disease controls and primary Sjogren's syndrome patients were non reactive to these sequences. Limited proteolysis of CRT with two major leucocyte serine proteases, elastase and cathepsin G, demonstrated that an N-terminal region of CRT is resistant to digestion. Interestingly, some of the epitopes with the highest reactivity belong to the fragments of the protein which bind to C1q and inhibit complement activation. Whether C1q association with CRT is a pathological or protective interaction between these two proteins is currently under investigation. PMID- 10792391 TI - Intra-articular IL-10 gene transfer regulates the expression of collagen-induced arthritis (CIA) in the knee and ipsilateral paw. AB - We studied the effects of local IL-10 application, introduced by a recombinant human type 5 adenovirus vector, in the mouse knee joint during the early phase of CIA. One intra-articular injection with the IL-10-expressing virus (Ad5E1mIL-10) caused substantial over-expression of IL-10 in the mouse knee joint, using virus dosages which did not induce distracting inflammation. High expression of IL-10 was noted for a few days, being maximal at day 1. One intra-articular injection of Ad5E1mIL-10 in the knee joints of collagen type II (CII)-immunized mice, before onset of CIA was noted, reduced the incidence of collagen arthritis in that knee. Of high interest, the protective effect of local IL-10 expression by Ad5E1mIL-10 was not restricted to the knee joint alone. The arthritis incidence in the ipsilateral paw was highly suppressed. In contrast, local IL-10 over expression was not effective when treatment was started after onset of CIA. Further analysis in the acute streptococcal cell wall-induced arthritis model revealed that local IL-10 over-expression markedly suppressed the production of tumour necrosis factor-alpha (TNF-alpha) and IL-1alpha, but had no significant effect on IL-1beta and IL-12 production in the inflamed synovium. These data indicate that local over-expression of IL-10 in the knee joint of mice regulates the expression of collagen arthritis, probably through down-regulation of TNF alpha. PMID- 10792394 TI - Autologous stem cell transplantation for malignancy: a systematic review of the literature. AB - A systematic review of the literature was undertaken to assess what published evidence is currently available to support the increasing use of autologous stem cell transplantation (ASCT), and to evaluate the published data with regard to the comparative cost of high-dose and conventional therapy. The review aimed to identify all published, randomized controlled trials (RCTs) comparing high-dose therapy (HDT) with ASCT versus conventional chemotherapy (CC) in acute lymphoblastic leukaemia, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, and breast, lung, testicular and ovarian cancer. The review also aimed to identify all studies that had compared the cost of the two treatment strategies. Reports were identified by systematic searches of Cancerlit, Embase and Medline, and handsearching of several conference proceedings. Where possible, pooled odds ratios (ORs) were calculated according to the fixed-effect model. A total of 18 randomized trials were identified in acute lymphoblastic leukaemia, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, and breast, lung and testicular cancer. Trials were generally small and no disease site had sufficient information to determine reliably whether high-dose therapy with autologous transplant is more effective than CC. Five studies were identified that compared the cost of the two treatments. These found the cost of HDT to be between one and four times higher than that of CC. Further randomized trials are required. Where appropriate, these should include economic assessment and assessments of long term toxicity. PMID- 10792395 TI - Molecular heterogeneity of the A3 subgroup. AB - The molecular characterization of the subgroup A3 remains unclear. Four unrelated A3 blood donors were studied. Family studies were possible in three of them. The A3 subgroup was defined by immunohaematological evaluation with four different commercially available serums. Exons VI and VII of the ABO gene, responsible for 91% of the catalytic active part of the glycosyltransferase, were amplified and subjected to direct sequencing. The results in all samples showed heterozygosity for the G261 deletion. In the A3 allele, the following associations were found: C467T mutation and 1060C deletion in one A3 blood donor and in another G829A and 1060C. In one case, only the 1060C deletion was demonstrated in the A3 allele. One blood donor presented the T646A and the G829A mutations in homozygosity. It was concluded that the A3 blood group is very heterogeneous at the molecular level. PMID- 10792393 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) but not granulocyte colony-stimulating factor (G-CSF) induces plasma membrane expression of proteinase 3 (PR3) on neutrophils in vitro. AB - The theoretical risk of triggering vasculitis resulting from administration of G CSF and GM-CSF to patients with anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitides (AAV), such as Wegener's granulomatosis (WG), who develop agranulocytosis due to cytotoxic therapy, is unknown. Since there is strong evidence that activation of polymorphonuclear neutrophils (PMN) induced by binding of ANCA to PR3 or myeloperoxidase (MPO) expressed on their plasma membrane is involved in the pathogenesis of systemic vasculitides (SV), we studied the surface expression of PR3 and MPO on PMN from healthy donors in response to G-CSF and GM-CSF in vitro by flow cytometric analysis. Increasing doses of G-CSF did not alter PR3 expression on either untreated or tumour necrosis factor-alpha (TNF-alpha)-primed donor PMN significantly. In contrast, GM CSF significantly increased PR3 membrane expression on both intact PMN and neutrophils primed with TNF-alpha. MPO expression was not significantly altered by either G-CSF or GM-CSF. In summary, these data demonstrate that GM-CSF, but not G-CSF, induces plasma membrane expression of PR3 on PMN in vitro. Since in AAV accessibility of the antigen (PR3 or MPO) to the antibody (ANCA) on the plasma membrane of PMN is thought to be essential for neutrophil activation by ANCA, the results of the present study suggest that administration of GM-CSF to patients with WG with neutropenia implies a definite theoretical risk of deterioration of vasculitis via this mechanism. PMID- 10792396 TI - Prevalence of hypochromia (without microcytosis) vs microcytosis (without hypochromia) in iron deficiency. AB - The usefulness of hypochromia (MCH or = 80 (fl) vs. counterparts with MCV < 80 fl (in the presence of MCH > or = 26 pg). Fifteen per cent of the 201 iron deficient subjects were also shown to have coexisting vitamin B12 deficiency. There was a comparable (16%) prevalence of this haematinic deficiency in the subgroup of 31 iron deficient patients with MCH < 26 pg in the presence of MCH > or = 80 fl. PMID- 10792397 TI - Total and differential leucocyte counts in infants at 2, 5 and 13 months of age. AB - Reference ranges for the total and differential leucocyte counts were determined from venous blood collected at 2, 5 and 13 months of age from a cohort of 112 healthy children of north European ancestry. At 2, 5 and 13 months, the ranges for neutrophils were found to be 0.7-4.7, 1.1-5.6 and 1.0-7.6 x 109/l, and for lymphocytes 3.3-10.5, 3.4-11.3 and 3.5-10.4 x 109/l, respectively. The upper limits for monocytes at each age were 1.2, 1.2 and 0.91 x 109/l, and for eosinophils 0.84, 1.0 and 0.88 x 109/l, respectively. Mean counts for all cell types, except monocytes, increased between 2 and 5 months of age. There was little change in mean counts between 5 and 13 months. Statistically significant correlations existed between the numbers of each cell type at 2 months of age, and were still present at 13 months between monocytes and each of the granulocyte series and between basophils and all other cell types. By comparison with older data these findings indicate a lower reference limit for neutrophils at 2 months of age, and a narrower range for this cell type at both 2 and 5 months of age. Reference ranges for lymphocytes and eosinophils are wider than indicated by some previous studies. PMID- 10792398 TI - Standardization of lymphocyte antibody binding capacity - a multi-centre study. AB - As quantitative flow cytometry is being increasingly used to characterize non malignant and malignant disorders, interlaboratory standardization becomes an important issue. However, the lack of standardized methods and process controls with predefined antibody binding capacity values, limits direct interlaboratory comparison. The present study has addressed these issues using a stable whole blood product and a standardized antigen quantification protocol. It was demonstrated that: (i) a standard technical protocol can result in a high degree of interlaboratory concordance; (ii) interlaboratory variation of less than 12% can be achieved for CD4 antibody binding capacity values; and (iii) stable whole blood can be used as a process control with predefined antibody binding capacity values. Furthermore, using such an approach, a normal range was established for CD3, CD4 CD8 and CD19. These antigens appear to be expressed in a hierarchical manner, a factor that could be used as a procedural quality control measure. PMID- 10792399 TI - Rapid detection of BCR/ABL and PML/RARA using fluorescence in situ hybridization in cytospin preparations. AB - Fluorescence in situ hybridization (FISH) is increasingly used as an adjunct to conventional cytogenetic analysis in the diagnosis of haematological malignancies and in monitoring minimal residual disease. FISH, however, is generally performed on slides prepared after short-term sample incubation and therefore, whilst faster than conventional cytogenetics, still requires a minimum of 2 days for a result to be obtained. A simplification of the FISH procedure is reported using uncultured cytospin preparations of bone marrow or peripheral blood for the rapid diagnosis of the BCR-ABL and PML-RARa gene rearrangements. It demonstrates that culturing has no effect on the ratio of normal to abnormal cells in the nondividing population. Data is presented from an analysis of 24 cases in whom unequivocal results were obtained in less than 12 h and in complete concordance with results obtained by conventional cytogenetics and/or interphase FISH. PMID- 10792400 TI - The determination of a reference range for new platelet parameters produced by the Bayer ADVIA120 full blood count analyser. AB - One hundred and twenty-two samples from normal volunteers were processed for full blood count analysis through the Bayer ADVIA120. The samples were simultaneously tested using the marker CD62P to establish the absence of platelet activation. Samples were tested at two time intervals, 20 min and 3.5 h, to simulate the testing practice for routine and urgent samples in our laboratory. From these data the mean and reference ranges, with 95% confidence limits, for platelet parameters were calculated for the ADVIA120. The time difference for some platelet parameters was significant. Hence two ranges have been reported. The age or smoking habit of the volunteer did not significantly affect platelet results but there was a statistically significant sex difference in some platelet parameters. PMID- 10792401 TI - Lupus anticoagulant in a patient with parvovirus B19 infection. AB - We report the case of a patient who presented with a prodromal illness suggestive of viral infection, subsequently confirmed as parvovirus B19, who had a lupus anticoagulant present. Her IgG anti-cardiolipin antibody was normal (7.0 units/ml) but her IgM anti-cardiolipin was elevated (55 units/ml). These later returned to normal. Parvovirus B19 infection may be associated with the presence of lupus anticoagulant. PMID- 10792402 TI - Severe skin rash in two consecutive patients treated with 2-chlorodeoxyadenosine for hairy cell leukaemia at a single institution. AB - Although hairy cell leukaemia was first described 40 years ago, it is only in the last decade that newer therapeutic agents have enabled effective treatment. The purine nucleoside analogue, 2-chlorodeoxyadenosine (2-CdA) is currently considered as first-line therapy with a very high rate of complete remission. Although adverse events with 2-CdA are increasingly recognized, severe cutaneous reactions have been reported rarely. We describe two consecutive patients treated with 2-CdA for hairy cell leukaemia who both suffered extremely severe cutaneous reactions, one of which was life-threatening. PMID- 10792403 TI - Natural killer (NK) cell leukaemia in a patient with a B cell non-Hodgkin's lymphoma. AB - We describe a 74-year-old woman with the diagnosis of natural killer (NK)-cell leukaemia and autoimmune pathology. Four years previously, a diffuse large B cell non-Hodgkin's lymphoma had been diagnosed and treated effectively. Although NK cell leukaemia has been thought to be a distinct highly aggressive clinicopathological entity, our case shows no further evolution at the present time. As far as we know, this association has not been previously described in the literature. PMID- 10792404 TI - Inverse relation between plasma G-CSF levels and neutrophil counts in a patient with autoimmune neutropenia treated with G-CSF. AB - The pharmacokinetic characteristics of granulocyte colony-stimulating factor (G CSF) appears to be related to the neutrophil count. We report the case of an 81 year-old male with acquired antibody induced neutropenia treated with G-CSF. This produced a rapid increase in the neutrophil count which appeared to be associated with diminished trough plasma G-CSF levels. Our data appears to indicate that mature neutrophils may play a part in the clearance of G-CSF from plasma. PMID- 10792405 TI - Association of B-chronic lymphocytic leukaemia and T-large granular lymphocyte leukaemia. PMID- 10792406 TI - Identification of a new Bruton's tyrosine kinase (BTK) mutation associated with a mild phenotype in a child with X-linked agammaglobulinemia (XLA). AB - This study reports a new X-linked agammaglobulinemia (XLA) mutation and its phenotypic features in a 6(1/2)-year-old boy. Different clinically defined subtypes of XLA may exist according to different genetic alterations and to other defect signalling molecules or pathways of B cell maturation. PMID- 10792407 TI - Successful high dose therapy for relapsed mediastinal large B cell lymphoma following surgical repair of anterior chest wall defect. AB - We describe a man with relapsed large B cell mediastinal lymphoma and associated infected large anterior chest wall defect who required high dose salvage therapy for his underlying disease. An initial mediastinotomy wound, associated with recurrent sepsis, had developed into an abscess, then fistula and eventually a large anterior chest wall defect. Safe use of salvage chemotherapy required reconstructive surgery consisting of a pedicled muscle flap. The subsequent high dose chemotherapy was carried out without complications and 15 months later the patient is alive and well. PMID- 10792408 TI - Classification and diagnostic criteria of the erythrocytoses. PMID- 10792409 TI - Effect of Qi-training on blood pressure, heart rate and respiration rate. AB - To examine the physiological effects of Korean traditional Qi-training, we investigated the changes in blood pressure, heart and respiratory rates before, during and after ChunDoSunBup (CDSB) Qi-training. Twelve normal healthy CDSB Qi trainees (19-37 years old; trained for 1.3 +/- 0.2 years; 9 men and 3 women) volunteered to participate in this study. Heart rate, respiratory rate, systolic blood pressure and rate-pressure product were significantly decreased during Qi training. From these results, we suggest that CDSB Qi-training has physiological effects that indicate stabilization of cardiovascular system. PMID- 10792410 TI - Vasoactive neuroendocrine responses associated with tolerance to lower body negative pressure in humans. AB - The purpose of this investigation was to test the hypothesis that peripheral vasoconstriction and orthostatic tolerance are associated with increased circulating plasma concentrations of noradrenaline, vasopressin and renin angiotensin. Sixteen men were categorized as having high (HT, n=9) or low (LT, n=7) tolerance to lower body negative pressure (LBNP) based on whether the endpoint of their pre-syncopal-limited LBNP (peak LBNP) exposure exceeded -60 mmHg. The two groups were matched for age, height, weight, leg volume, blood volume and maximal oxygen uptake, as well as baseline blood volume and plasma concentrations of vasoactive hormones. Peak LBNP induced similar reductions in mean arterial pressure in both groups. The reduction in leg arterial pulse volume (measured by impedance rheography), an index of peripheral vascular constriction, from baseline to peak LBNP was greater (P<0.05) in the HT group (-0.041 +/- 0.005 ml 100 ml-1) compared to the reduction in the LT group (-0. 025 +/- 0.003 ml 100 ml-1). Greater peak LBNP in the HT group was associated with higher (P<0.05) average elevations in plasma concentrations of vasopressin (pVP, Delta=+7.2 +/- 2.0 pg ml-1) and plasma renin-angiotensin (PRA, Delta=+2.9 +/- 1.3 ng Ang II ml-1 h-1) compared to average elevations of pVP (+2.2 +/- 1.0 pg ml-1) and PRA (+0.1 +/- 0.1 ng Ang II ml-1 h-1) in the LT group. Plasma noradrenaline concentrations were increased (P<0.05) from baseline to peak LBNP in both HT and LT groups, with no statistically distinguishable difference between groups. These data suggest that the renin-angiotensin and vasopressin systems may contribute to sustaining arterial pressure and orthostatic tolerance by their vasoconstrictive actions. PMID- 10792411 TI - Physical activity, aerobic capacity and selected markers of oxidative stress and the anti-oxidant defence system in healthy active elderly men. AB - The relationship of oxidative stress and the anti-oxidant defence system to maximal oxygen consumption (VO2max) and habitual physical activity was assessed in 26 elderly men (71.0 +/- 4.2 years) and compared to that of 12 young men (22.1 +/- 5.1 years). Physical activity was assessed by a questionnaire. Malondialdehyde (MDA), plasma total anti-oxidant status (TAS), the levels of red blood cell (RBC) superoxide dismutase (SOD) and glutathione peroxidase (GPX), as well as serum GPX activities were determined under resting conditions. The older and young men had similar TAS and RBC SOD, while MDA, RBC GPX and plasma GPX were higher, and RBC SOD/GPX ratio was significantly lower in the older men. Neither MDA nor anti-oxidants were associated with any of the physical activity/aerobic capacity measures in the elderly men. We conclude that in healthy elderly men with a good nutritional status, indicators of the anti-oxidant defence system are not lower in comparison with young men. Increased RBC and plasma GPX coupled with a high level of lipid peroxidation marker may indicate an adaptation of anti oxidant defences to sustained oxidative stress. Furthermore, the results of the present study suggest that the level of habitual physical activity and aerobic capacity have no major influence on the resting balance between radical generation and blood anti-oxidant potential in healthy older men. PMID- 10792412 TI - Study of the effects of various transcutaneous electrical nerve stimulation (TENS) parameters upon the RIII nociceptive and H-reflexes in humans. AB - Despite over two decades of clinical use, the neurophysiological and anti nociceptive effects of transcutaneous electrical nerve stimulation (TENS) have yet to be definitively described. The current study was designed to examine the effect of TENS on the RIII nociceptive reflex elicited in healthy human subjects; the H-reflex was measured concomitantly to monitor changes in alpha-motoneuron excitability. Following approval from the university's ethical committee, 50 healthy human volunteers (25 male and 25 female) participated in the study. The subjects ranged in age from 18 to 30 years (mean 22, SD 3). Subjects were randomly allocated equally to a control group or one of four TENS groups. In the TENS groups, stimulation was applied for a total of 15 min over the sural nerve in the left leg. Ipsilateral RIII and H-reflexes were recorded five times during the 45 min experimental period. In addition, subjects also rated pain associated with the RIII reflex using a computerized visual analogue scale (VAS). Statistical analysis using two-way repeated-measures ANOVA showed no differences between groups for H-reflex, RIII reflex nor VAS data. These results suggest that TENS does not significantly affect either of the two reflexes, at least using the parameters and application time in the current study. PMID- 10792413 TI - Low-frequency respiratory rhythms in infants during the first six months of life. AB - The aim of the study was to investigate characteristics of low-frequency components in respiration. Sixteen healthy term infants were examined from the first day up to the 6th month of life. The respirogram, instantaneous respiratory frequency and respiratory amplitude of undisturbed segments of quiet sleep phases and periodic breathing (PB) were analysed via fast Fourier transformation. The peak frequency (PF) in the low-frequency range (0.04-0.2 Hz) was determined. PF for PB ranged from 0.056 to 0.1 Hz. Further, low-frequency rhythms (LFR) of the respirogram, which were stable during the recordings as well as during development, were found ranging from 0.045 to 0.067 Hz. The LFR of the respirogram is correlated with rhythmic changes in the relationship between inspiratory and expiratory amplitudes. The frequency of the LFR was significantly lower than that of the PB. The data indicate that LFR and PB are low-frequency respiratory rhythms which are separately controlled and perform independently. PMID- 10792414 TI - Effect of nasal air temperature on FEV1 and specific airways conductance. AB - We investigated whether changes in nasal air temperature affect specific airway conductance (SGaw) and one second forced expiratory volume (FEV1) in 10 asthmatic patients with a history of cold-sensitive asthma and eight healthy subjects. An air-stream (0.6 l s-1) of -15 degrees C, +22 degrees C or +37 degrees C was blown into both nostrils during a Valsalva manoeuvre. Each provocation consisted of 10 puffs of air, each of 15 s duration, at 1 min intervals. Before and at regular intervals after the provocations, SGaw and FEV1 were determined. In asthmatics, after cold air provocation, SGaw fell 23% (P<0.01) and FEV1 8% (P<0.01). After the warm air provocations, SGaw rose 15% (P<0.01) and FEV1 6% (P<0.01). After the ambient air provocations, no significant changes occurred in SGaw or FEV1. In the healthy subjects, the nasal provocations caused no significant changes in lung function. The present study shows that cold air in the nose causes a slight obstruction and warm air possibly a slight dilatation of the lower airways in patients with a history of cold-sensitive asthma but not in healthy subjects. PMID- 10792415 TI - The relation between ejection fraction and mitral annulus motion before and after direct-current electrical cardioversion. AB - Mitral annulus motion (MAM) and the relation between left ventricular ejection fraction (EF) and MAM has been shown to differ between patients with sinus rhythm and patients with atrial fibrillation. However, it has not been investigated how the relation between EF and MAM changes on direct-current (DC) electrical cardioversion to sinus rhythm. Therefore, 31 consecutive patients on the waiting list for DC electrical cardioversion were examined by echocardiography before DC electrical cardioversion, and those who maintained sinus rhythm (13 patients) were examined again 4-8 weeks after cardioversion. The conversion factor (CF) (ratio EF/MAM) decreased from 8.4 +/- 1.7 before to 5.8 +/- 0.8 SD after cardioversion (P<0.001). The EF increased slightly (P<0.05) but the MAM had a much greater increase (P<0.001), resulting in the decrease in CF. There was no significant difference in CF between patients after cardioversion and age- and gender-matched control patients with sinus rhythm, indicating that CF is normalized or almost normalized 4-8 weeks after cardioversion. This indicates that when MAM is used for investigation of the left ventricular function, and the function is expressed as EF, the same CF as in other patients with sinus rhythm can be used 4-8 weeks after DC electrical cardioversion. PMID- 10792416 TI - Isotonic dynamometry for the assessment of power and fatigue in the knee extensor muscles of females. AB - Impairments in muscle power production and recovery following short-duration intense activity could lead to decreased performance and risk of injury. We developed a power test for the knee extensor muscles using torque-velocity testing and moderate isotonic loads. Twenty-eight female volunteers performed three maximal efforts at each of four isotonic loads (27.1, 40.6, 54.2 and 67.8 N. m). If the calculated regression line for the torque-velocity data had an r2 >/= 0.95 (i.e. an acceptable test), maximal power (408 +/- 56 W) was computed from the data. Immediately after torque-velocity testing, the subjects repeated maximal effort knee extensions with 33.9 N. m for three bouts of 15 repetitions with 15 s of rest to produce muscle fatigue, defined as a decrease in power output during isotonic exercise. After a 4 min rest, the torque-velocity test was repeated and power calculated (345 +/- 48 W). For the group, the recovery of maximal power after the fatigue protocol was 85%. The extremes were represented by one subject who recovered only 70% of her maximal power and another who recovered completely (>98%). Physiological differences in muscle power following repeated exercise could have an impact on the outcome of therapeutic interventions for sports injuries, fatigue syndromes and occupational over-use conditions. PMID- 10792417 TI - Autonomic cardiovascular regulation in patients with obstructive sleep apnoea: a study based on spectral analysis of heart rate variability. AB - The aim of this study was to assess the function of the autonomic nervous system in patients with obstructive sleep apnoea syndrome (OSAS). The study was designed as a cross-sectional case-control study. Fifty-one patients were included, and the findings were compared with those in 66 controls. Spectral analysis of heart rate variability (HRV) during supine rest, during controlled breathing and after tilting was performed in each patient and control case. The patients performed overnight sleep recordings the night before the HRV recordings. Individuals with an apnoea-hypopnoea index (AHI) above 20 were regarded as OSAS patients and those with AHI lower than 20 as snorers. Differences in HRV and blood pressure between patients and controls were analysed by multiple linear regression with age, body mass index and sex as independent variables. During free and controlled breathing there was a significant decrease in indices reflecting vagal modulation, indicating parasympathetic dysfunction in OSAS patients compared with controls. The mid-frequency component was also significantly reduced in OSAS patients after tilting but not in the lying position. This may be related to the parasympathetic dysfunction, but could also indicate a decreased sympathetic reserve capacity. We found no significant relation between AHI and indices of vagal modulation in the patient group. Our findings show an autonomic dysfunction in patients with OSAS. The dysfunction involves the parasympathetic system, and may be related to the increased cardiovascular mortality and malignant arrhythmia described in OSAS. PMID- 10792418 TI - Distribution of interstitial fluid pressure and fluid volumes in hind-limb skin of rats: relation to meridians? AB - To determine the distribution of interstitial fluid pressure (Pi) and volume (Vi), and to relate the distribution of these parameters to the distribution of potential meridians located by measurement of electrical impedance, we measured Pi, extracellular fluid (Ve) and plasma volumes (Vp) in 14 pre-defined skin areas, 2 x 2 mm, and in concave and convex regions on the hind-limb and groin of rats in control conditions. Pi was measured with sharpened glass capillaries connected to a servo-controlled counter-pressure system, while Ve and Vp were determined as the extravascular distribution spaces of 51Cr-EDTA and 125I-human serum albumin, respectively. Vi was calculated as Ve - Vp, and Vw as the difference between skin wet and dry weight. Grand mean Pi averaged -0.81 mmHg (SD 0.83, n=95). Pi in skin was significantly higher in lateral and medial parts of the medial aspect of hind-limb compared to pressures in the intermediate area (P<0.05). Pressures in the concave groin and the convex knee area were more negative and positive, respectively, than in the flat intermediate central hind limb area. There was a significantly higher Vi (P<0.05) and Vw (P<0.05) in the lateral side than that in the medial side. Vp was higher medially and laterally than in the intermediate area (P<0.05 for both comparisons), and correlated positively and significantly with Pi (r=0.66, P<0.05). No correlation was found between Pi and electrical impedance. The study suggests that the distribution of Pi, Vi, Vp and Vw is heterogeneous in hind-limb skin at a macroscopic level without obvious relations to potential meridians. PMID- 10792419 TI - Common polymorphisms in the CTLA-4 and CD28 genes at 2q33 are not associated with asthma or atopy. AB - Recently, genetic linkage of the chromosomal region 2q33 with asthma has been shown. The genes coding for CD28 and CTLA-4 have been localized to this chromosomal region. CD28 and CTLA-4 have been shown to be involved as an important costimulatory signal in the regulation of allergic inflammation and TH2 cytokine production, and thus both genes are good candidate genes for asthma and atopy. Two common polymorphisms in the CTLA-4 gene and one polymorphism in the CD28 gene found by single-strand conformation polymorphisms (SSCP) analysis and direct genomic sequencing were tested for association with asthma and atopy phenotypes in a population of 260 largely atopic children and young adults. No association was found between any of the three polymorphisms and asthma or atopy phenotypes. The newly described common CD28 polymorphism is situated in the third intron of the gene. We conclude that neither gene is likely to exert a major influence on the development of asthma or atopy in our population. However, it might prove useful to test for association of these polymorphisms with asthma in populations recruited through asthmatic but not necessarily atopic individuals. PMID- 10792420 TI - A polymorphic major histocompatibility complex class II-like locus maps outside of both the chicken B-system and Rfp-Y-system. AB - Chickens have two major regions encoding major histocompatibility complex (MHC) class Ialpha genes and MHC class IIss genes, the serological and functional B system and the Rfp-Y-system. Recently, they have been shown to assort in a genetically independent way although still located on the same microchromosome. Moreover, the monomorphic MHC class IIalpha gene maps at a third locus located 5 cM from the nearest class IIss genes, located in the B-system (Kaufman et al., 1995). A pedigree family was studied in three generations in order to assign MHC class IIss restriction fragments observed in Southern blot analyses to either the B-system, the Rfp-Y-system or the B-Lalpha locus. In this study, we demonstrate by classical genetic testing of chickens within this fully pedigreed family the existence of an MHC class II-like polymorphic restriction fragment that segregates independently of the B-system, the Rfp-Y-system and of the B-Lalpha locus. PMID- 10792421 TI - Intercellular adhesion molecule-1 gene polymorphisms in Behcet's disease. AB - Intercellular adhesion molecule-1 (ICAM-1) gene polymorphisms have been implicated in the susceptibility to inflammatory diseases, including multiple sclerosis and inflammatory bowel disease. The expression of both soluble and tissue ICAM-1 is increased in Behcet's disease (BD) but the contribution of ICAM 1 gene polymorphisms to this disease remains unknown. Associations with BD have been reported for genes within the MHC, including HLA-B51, TNF and MICA, but the role of non-MHC genes in BD remains largely unexplored. We have investigated the frequency of the R/G 241 and K/E 469 ICAM-1 gene polymorphisms in 83 patients with BD disease and 103 healthy controls, all of Palestinian and Jordanian descent, and demonstrated an association between BD and the ICAM-1 E469 allele (Pc = 0.046, OR = 2.1). Among patients, no association was found between the presence of ocular disease and ICAM-1 polymorphisms. While the functional correlate of this polymorphism remains unclear, this finding indicates that a genetic polymorphism in the ICAM-1 gene domain, which is independent of the MHC, may contribute to disease. PMID- 10792422 TI - CTLA4 exon 1 dimorphism in bullous and cicatricial pemphigoid. AB - The human cytotoxic T-lymphocyte antigen 4 (CTLA4) gene encodes proteins regulating the immune response. The polymorphism of this gene is associated with some autoimmune diseases. In this study, we analysed the distribution of the dimorphisms of exon 1 (+ 49 A/G) in bullous pemphigoid (BP) and cicatricial pemphigoid (CP), two types of autoimmune bullous skin diseases that occur in elderly people. The frequency of the exon 1 A-G genotype was marginally decreased in patients (36.4%; n = 55) compared with controls (52.8%, n = 53), but the results were not statistically significant (P = 0.09). PMID- 10792424 TI - The effect of geographic origin on the frequency of HLA antigens and their association with ageing. AB - The association between HLA antigens and ageing is not clear. Ageing in women was associated with B40 and DR5 in a recent study, but other studies yielded conflicting results. In none of the studies, however, did the young and elderly samples originate from the same homogeneous population. Homogeneity is dependent on geographic origin. The aim of this study was to investigate whether differences in geographic origin between age groups could explain the age associated differences in the frequencies of B40 and DR5. The authors used the new design of a 'birth-place-restricted comparison' in which the origin of all subjects was ascertained. The total study population comprised 1010 young women aged 25-40 years and 660 elderly women aged 85 years and older. The 'birth-place restricted comparison' included 66 young and 285 elderly women from one geographic area (Leiden, the Netherlands). Men were not included because ageing in men was not associated with HLA antigens in a recent study. In the total population, the frequency of B40 in young women of different origin varied between 16 and 28%, and the frequency of DR5 between 11 and 23%. Similar differences were observed in the elderly women. In the 'birth-place-restricted comparison', the frequency of B40 was 15% in the young women and 11% in the elderly women (difference 4%, 95% confidence interval, -5 to 13%). The frequency of DR5 was 20% in the young women, and 28% in the elderly women (difference 8%, 95% confidence interval, -4 to 19%). Thus, marked differences in HLA antigen frequency were found between populations of various geographic origins. Definition and ascertainment of the target population are therefore necessary in genetic studies of ageing. In such a 'birth-place-restricted comparison', the authors confirmed that ageing in women was negatively associated with HLA-B40 and positively associated with HLA-DR5. PMID- 10792423 TI - Analysis of the phenotypic distribution of HLA class I and class II in atopic and non-atopic asthma patients. AB - In several studies the HLA system has been implicated in the development of asthma, but the importance of the associations between HLA genes and asthma remains unclear. The aim of this study was to determine the HLA class I and II phenotypic frequencies in a population of asthmatics, and to analyse the relationship between these phenotypes and any type of asthma. We typed HLA class I and II antigens in a series of 189 asthmatic individuals (102 atopic and 87 non atopic), and in a control population of 150 unrelated healthy Caucasoid donors. When the HLA phenotypic frequencies were compared, no statistical differences were found. Therefore, no definitive HLA association could be established with atopic or non-atopic asthma in the studied population. Abbreviations AA, atopic asthma; FEV1, forced expiratory volume in 1 s; NAA, non-atopic asthma; PCR, polymerase chain reaction; SSOP, sequence-specific oligonucleotide probes; SPT, skin prick test. PMID- 10792425 TI - HLA antigen and gene frequencies in Eskimos of East Greenland. AB - A population of 78 Ammassallik Eskimos was tested for HLA-A, B, Cw, DR and DQ specificities using serological techniques and HLA-Cw, DRB, DQB1 and DPB1 using three different genomic techniques. The application of two new genomic techniques for HLA-Cw (PCR-SSP phototyping) and HLA-DPB1 (PCR-RLFP) typing confirmed serological results and gave a higher resolution, with more heterozygotes than previously found. High gene frequencies of HLA-A24 (0.80), B51 (0. 21), B61 (0.30), B62 (0.21), Cw*0303 (0.27), Cw*0304 (0.51), DRB1*0401 (0.45), DRB1*1402 (0.24), DQ7 (0.88), DPB1*0201 (0.18), DPB1*0401 (0.40) and DPB1*0402 (0.30) were found. A limited number of alleles were found at all HLA loci. The Ammassallik Eskimos of East Greenland are genetically homogenous with little, if any, admixture with European populations. This contrasts with other Greenlandic populations formerly tested. The pattern of alleles indicates a close genetic relationship with other Eskimo populations. PMID- 10792426 TI - Molecular cloning and sequencing of a cDNA encoding the feline cytotoxic T lymphocyte-associated protein 4 (CTLA4) homologue. AB - The feline CTLA4 cDNA encodes a transmembrane protein which shares 87.9% sequence identity with the human CTLA4 molecule. The cytoplasmic region of the feline CTLA4 is identical to that of humans and mice, which suggests conserved function(s) such as the regulation of cell-surface expression of this molecule. PMID- 10792428 TI - Nomenclature for factors of the HLA system, update October/ November 1999. PMID- 10792427 TI - Characterization of a rare sty I polymorphism in exon 1C of the human interleukin 1 receptor type I (IL-1RI) gene. PMID- 10792429 TI - Nomenclature for factors of the HLA system, update December 1999. PMID- 10792430 TI - Regulation of GluR2 promoter activity by neurotrophic factors via a neuron restrictive silencer element. AB - The AMPA glutamate receptor subunit GluR2, which plays a critical role in regulation of AMPA channel function, shows altered levels of expression in vivo after several chronic perturbations. To evaluate the possibility that transcriptional mechanisms are involved, we studied a 1254-nucleotide fragment of the 5'-promoter region of the mouse GluR2 gene in neural-derived cell lines. We focused on regulation of GluR2 promoter activity by two neurotrophic factors, which are known to be altered in vivo in some of the same systems that show GluR2 regulation. Glial-cell line derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) both induced GluR2 promoter activity. This was associated with increased expression of endogenous GluR2 immunoreactivity in the cells as measured by Western blotting. The effect of GDNF and BDNF appeared to be mediated via a NRSE (neuron-restrictive silencer element) present within the GluR2 promoter. The response to these neurotrophic factors was lost upon mutating or deleting this site, but not several other putative response elements present within the promoter. Moreover, overexpression of REST (restrictive element silencer transcription factor; also referred to as NRSF or neuron restrictive silencer factor), which is known to act on NRSEs in other genes to repress gene expression, blocked the ability of GDNF to induce GluR2 promoter activity. However, GDNF did not alter endogenous levels of REST in the cells. Together, these findings suggest that GluR2 expression can be regulated by neurotrophic factors via an apparently novel mechanism involving the NRSE present within the GluR2 gene promoter. PMID- 10792431 TI - Response properties of PMLS and PLLS neurons to simulated optic flow patterns. AB - The processing of optic flow information has been extensively investigated in the medial superior temporal area (MST) of the macaque. In the cat, the posteromedial area and the posterolateral area in the lateral suprasylvian cortex (PMLS and PLLS, respectively) have been suggested as likely participants according to their direction preferences to moving objects. In the present study, 203 PMLS and 123 PLLS neurons were tested with simulated optic flow patterns composed of random dots (including expansion and contraction, clockwise and counter-clockwise rotation, and translation) and moving bar stimuli. About 90% of the neurons were found to be excited by the optic flow stimuli and most of them were multiple responsive to different flow patterns. Only 20-25% of the cells were selective to different optic flow modes, and in general, the direction preference was fairly modest. The selective cells showed stronger directionality to both flow field and moving bar than nonselective cells. However, the optic flow response properties in the PMLS and PLLS were not well correlated with the direction preference to moving bars. In accordance with previous findings, the PMLS was analogous to the middle temporal area of the macaque in many respects. As for the PLLS cells, they were sensitive to fewer types of stimuli, but responded better and more selectively to radial motion. All these results suggest that the two lateral suprasylvian areas are unlikely to be specialized for the analysis or discrimination of different flow patterns, but may play some kind of relay role in optic flow information processing. PMID- 10792432 TI - Developmental expression of amphiphysin in the retinotectal system of the chick: from mRNA to protein. AB - The role of amphiphysin in clathrin-mediated endocytosis of synaptic vesicles is well established. However, it is still uncertain if the protein is also involved in developmental mechanisms, e.g. axon outgrowth and synapse formation. To investigate the developmental changes in the expression of amphiphysin we used the retinotectal system of the chick, a highly ordered and easily accessible primary neuronal pathway. Reverse transcription polymerase chain reaction (RT PCR) of total RNA from chick retina and tectum revealed first transcripts for amphiphysin, dynamin and synaptotagmin at embryonic day 5 (E5) for both regions. Surprisingly, Western blots of the retina revealed an increase of protein expression for amphiphysin only after E11 in the retina and the tectum. Immunofluorescence for amphiphysin was not detectable before E10 in the developing chick retina, while other presynaptic proteins like synaptotagmin showed already intense signals in the inner and outer plexiform layers. Subsequently, amphiphysin immunoreactivity follows the expression of synaptotagmin and synaptic vesicle protein 2 (SV2) as seen in the retina and the tectum, and exhibits the same staining as the other proteins in the mature chick brain. Ultrastructural data revealed for the first time that amphiphysin is not only limited to conventional synapses but is also abundant in retinal ribbon terminals. Taken together our data reveal that: (i) there is a developmental delay between mRNA transcription and protein expression for key proteins involved in endocytosis; (ii) amphiphysin gets upregulated after synapse formation; and (iii) amphiphysin is present in the synaptic vesicle cycle in retinal ribbon synapses. PMID- 10792433 TI - Neurofilament protein distribution in the macaque monkey dorsolateral premotor cortex. AB - Regional and laminar distribution patterns of neurofilament proteins in the dorsolateral premotor cortex (PMd) were studied with monoclonal antibody SMI-32 in five adult macaque monkeys and compared with the cytoarchitectonical features of the PMd. Our goal was to reveal whether the increasing functional diversity of the PMd which electrophysiological studies have unravelled over the last years is reflected on a structural level by differences in the neurochemical phenotype. Differences in size, shape and packing density of immunopositive layer III and V pyramidal cells define areas much more clearly than do differences in cytoarchitecture. The PMd can be subdivided into a rostral and a caudal part at a level slightly anterior to the genu of the arcuate sulcus. The extent of these two areas matches the two cytoarchitectonically defined areas F7 and F2, respectively. Within area F2, differences in layer V immunoreactive neurons define a dorsal (F2d) and a ventral (F2v) region. The border between areas F2d and F2v lies at the superior precentral dimple and cannot be detected cytoarchitectonically in Nissl-stained sections. Neurofilament proteins are involved in the stabilization of the cytoskeleton of the axon and have been correlated with axonal size and conduction velocity of nerve fibres. This regional variability in the neurochemical phenotype of layer V within the caudal PMd may reflect a differential organization of the descending output from this part of the premotor cortex. It might also be related to differences in the motor control of voluntary arm and leg movements. PMID- 10792434 TI - Galanin influences the mechanosensitivity of sensory endings in the rat knee joint. AB - The aim of the present study was to examine the effect of galanin on group III and IV afferent nerve fibres (n = 53) innervating normal and acutely inflamed knee joints in rats. They responded to local mechanical stimulation, movements of the joint and i.a. injections of KCl close to the joint. Single i.a. bolus injections of galanin (0.1 mM, 0.2 mL) caused no direct responses of the units. In normal and acutely inflamed joints, about half of the units did not change the responses to knee joint rotation. A significant reduction of the responses to noxious movements was found in approximately 40% of the units reaching a mean value of 57% in normal joints and 70% in inflamed joints compared with control movements. In approximately 10% the responses increased to 143% in normal joints and 120% in inflamed joints. Injection of a galanin receptor antagonist (M35) doubled the responses to noxious movements in 36% of the units in normal joints and reduced it in 18% to 86% of the control movements, indicating a tonic release and influence on the mechanosensitivity of a proportion of primary afferents by galanin. In conclusion, these data further support the hypothesis that the mechanosensitivity of fine afferent nerve fibres is regulated by a mixture of different substances being released into the innervated tissue. Besides the action of several pro-inflammatory peptides there seems to exist a tonic inhibitory system. PMID- 10792435 TI - MPTP selectively induces haem oxygenase-1 expression in striatal astrocytes. AB - Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta with accompanying evidence of increased oxidative damage, deficits in mitochondrial function and iron deposition. Recently, haem oxygenase-1 levels were reported to be elevated in PD brains. Because this enzyme is involved in the response to oxidative stress and is critical for cellular haem and iron homeostasis, it could play a role in the pathogenesis of PD. Therefore, we investigated the expression of haem oxygenase isoform 1 (HO-1) in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. MPTP triggered a relatively rapid and persistent increase in HO-1 mRNA exclusively in the mouse striatum. In situ hybridization and immunohistochemistry showed HO-1 to be localized to striatal astrocytes. The induction of HO-1 by MPTP was blocked by selegiline and GBR-12909, indicating the protoxin had to be metabolized by monoamine oxidase B and taken up by dopaminergic neurons to exert its action in astrocytes. MPTP did not alter the expression of other enzymes of haem synthesis or degradation nor were the levels of mRNA for haem or iron-binding proteins changed. Thus, expression of HO-1 was not part of a cellular program involving haem biosynthesis or homeostasis. In addition, heat shock proteins were not induced by MPTP. Thus, MPTP elicited a selective transcriptional response in striatal astrocytes. This response appears to be mediated by molecules released from affected dopaminergic nerve terminals in the striatum acting upon neighbouring astrocytes. This signalling pathway and its potential relevance to PD are discussed. PMID- 10792436 TI - Neurotrophin 3 potentiates glutamatergic responses of IHC afferents in the cochlea in vivo. AB - Neurotrophins have traditionally been regarded as slow-acting signals essential for neuronal survival and differentiation. Recent studies with neuronal slices, cultures and nerve ending preparations have shown that neurotrophins generate acute changes in nerve activity. Among the secondary sensory cells are the inner hair cells (IHC) and taste buds, cells which express the neurotrophic factors necessary for the survival of their innervating neurons. If in these cells neurotrophins acutely affect the nerve activity of their afferent neurons, as in the central nervous system (CNS), this may have important functional implications for the corresponding sensory transduction processes. The neurotrophin NT-3 has been reported to be expressed in IHCs. We chose an in vivo application system for the microiontophoretic supply of NT-3 in the subsynaptic region of the IHC. The effect of NT-3 on spontaneous and evoked afferent cochlear nerve activities in adult guinea pig inner ear was studied. We observed that NT-3 rapidly increases the spontaneous and glutamate-evoked firing rate of IHC afferents. Moreover, firing induced by both N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) were specifically enhanced during the presence of NT-3, a process which was selectively blocked by the tyrosine kinase receptor inhibitor K252a. Because we localized NT-3 mRNA not only in IHCs but also in the spiral ganglion, we propose that similar to other sensory systems, afferent and autocrine neurotrophin activities may be responsible for survival of cochlear neurons. In addition, NT-3 in IHCs may operate as a signal-dependent, intrinsic neuromodulator and/or neuroprotector. PMID- 10792437 TI - The distribution and origin of the calretinin-containing innervation of the nucleus accumbens of the rat. AB - The nucleus accumbens of the rat consists of several subregions that can be distinguished on the basis of histochemical markers. For example, the calcium binding protein calbindin D28k is a useful marker of the core compartment of the nucleus accumbens. Calretinin, another calcium-binding protein, is found in a dense fibre plexus in the accumbal shell and septal pole regions. The source of the accumbal calretinin innervation is not known. We examined the distribution of calretinin in the nucleus accumbens and used tract-tracing and lesion methods to determine the source of this calretinin innervation. Intense calretinin immunoreactivity was present in the medial shell, but the density of calretinin axons diminished sharply in the ventrolateral shell. Regions of dense calretinin immunostaining and those areas with calbindin-like immunoreactive cell bodies were generally segregated in the nucleus accumbens, although some overlap in the transition region between the core and shell was seen. Small clusters of calretinin-immunoreactive fibres were seen in the core, where they were restricted to calbindin-negative patches. Injections of the anterograde tracer biotinylated dextran amine into the paraventricular thalamic nucleus labelled fibres in calretinin-rich regions of the accumbens. Conversely, injections of Fluoro-gold into the accumbal shell retrogradely labelled numerous cells in the paraventricular thalamic nucleus that were calretinin-immunoreactive. Electrolytic lesions of the paraventricular thalamic nucleus reduced calretinin levels in the shell by approximately 80%. These data indicate that the calretinin innervation of the nucleus accumbens is derived primarily from the thalamic paraventricular nucleus, and marks accumbal territories that are largely complementary to those defined by calbindin. PMID- 10792439 TI - Progesterone regulation of GABAA receptor plasticity in adult rat supraoptic nucleus. AB - Marked plasticity in GABAA receptor signalling occurs in adult oxytocin neurons of the supraoptic nucleus (SON) through the modulation of GABAA receptor alpha subunits during pregnancy. The present studies were undertaken to examine the potential mechanisms underlying this plasticity. In vivo microdialysis experiments in conscious rats revealed that no significant changes in extracellular GABA concentrations occurred within the SON over the last two days of pregnancy and the time of parturition itself. In situ hybridization studies examined the effects of gonadal steroid manipulation upon the GABAA receptor subunits expressed by SON neurons (alpha1, alpha2, beta2 and gamma2 subunits) and demonstrated that cellular levels of the alpha1 subunit were increased following 8 days oestrogen and progesterone treatment. Estrogen alone or allopregnanolone, the progesterone derivative, had no effect on alpha1 subunit mRNA expression in the SON. Immunocytochemical experiments demonstrated progesterone receptors in many neural populations but not within the SON of late pregnant rats. These studies indicate that alterations in endogenous GABA release within the SON are unlikely to be responsible for the GABAA receptor plasticity exhibited by oxytocin neurons in late pregnancy. Rather, data demonstrate that the fluctuating concentrations of progesterone during pregnancy act indirectly on SON neurons to modulate alpha1 subunit mRNA expression. Together, these experiments provide evidence for the ligand-independent induction of GABAA receptor plasticity in the adult brain by progesterone. PMID- 10792438 TI - Distinct expression patterns of eph receptors and ephrins relate to the structural organization of the adult rat peripheral vestibular system. AB - Eph receptors and their ligands, termed ephrins, have been implicated in axon guidance, neuron-target interactions, regional compartmentalization, and synaptic functions in nervous systems. These activities of the Eph family molecules prompted us to investigate whether these molecules play roles in the maintenance, regeneration and plasticity in the mature peripheral vestibular system. Using reverse transcription-polymerase chain reaction (RT-PCR), Western blot and immunocytochemical analyses, we identified distinct and reciprocal expression patterns of full-length isoforms of EphA5, EphA6, EphA7, EphB1, ephrin-A2 and ephrin-B1 that correlated with structural features of the peripheral vestibular system in adult rats. All of the Eph receptors and ephrins examined were localized in the cell bodies of vestibular ganglion neurons in vivo and in vitro, and were readily detected in their outgrowing neurites in vitro. In the utricle, these molecules were localized in distinct cellular and subcellular compartments corresponding to discrete features of utricular afferent innervation, e.g. defasciculation, branching and synapse formation. Taken together, these results identify the Eph receptors and ephrins as candidate molecular substrates for defining some aspects of the structural organization of the adult peripheral vestibular system. PMID- 10792440 TI - Characterization of calcium currents in functionally mature mouse spinal motoneurons. AB - Motoneurons integrate synaptic input and produce output in the form of trains of action potentials such that appropriate muscle contraction occurs. Motoneuronal calcium currents play an important role in the production of this repetitive firing. Because these currents change in the postnatal period, it is necessary to study them in animals in which the motor system is 'functionally mature', that is, animals that are able to weight-bear and walk. In this study, calcium currents were recorded using whole-cell patch-clamp techniques from large (> 20 microm) ventral horn cells in lumbar spinal cord slices prepared from mature mice. Ninety percent (nine out of 10) of the recorded cells processed for choline acetyltransferase were found to be cholinergic, confirming their identity as motoneurons. A small number of motoneurons were found to have currents with low voltage-activated (T-type) characteristics. Pharmacological dissection of the high-voltage-activated current demonstrated omega-agatoxin-TK- (P/Q-type), omega conotoxin GVIA- (N-type), and dihydropyridine- and FPL-64176-sensitive (L-type) components. A cadmium-sensitive component of the current that was insensitive to these chemicals (R-type) was also seen in these cells. These results indicate that the calcium current in lumbar spinal motoneurons from functionally mature mice is mediated by a number of different channel subtypes. The characterization of these calcium channels in mature mammalian motoneurons will allow for the future study of their modulation and their roles during behaviours such as locomotion. PMID- 10792441 TI - Dendritic L-type calcium currents in mouse spinal motoneurons: implications for bistability. AB - The intrinsic properties of mammalian spinal motoneurons provide them with the capability to produce high rates of sustained firing in response to transient inputs (bistability). Even though it has been suggested that a persistent dendritic calcium current is responsible for the depolarizing drive underlying this firing property, such a current has not been demonstrated in these cells. In this study, calcium currents are recorded from functionally mature mouse spinal motoneurons using somatic whole-cell patch-clamp techniques. Under these conditions a component of the current demonstrated kinetics consistent with a current originating at a site spatially segregated from the soma. In response to step commands this component was seen as a late-onset, low amplitude persistent current whilst in response to depolarizing-repolarizing ramp commands a low voltage clockwise current hysteresis was recorded. Simulations using a neuromorphic motoneuron model could reproduce these currents only if a noninactivating calcium conductance was placed in the dendritic compartments. Pharmacological studies demonstrated that both the late-onset and hysteretic currents demonstrated sensitivity to both dihydropyridines and the L-channel activator FPL-64176. Furthermore, the alpha1D subunits of L-type calcium channels were immunohistochemically demonstrated on motoneuronal dendrites. It is concluded that there are dendritically located L-type channels in mammalian motoneurons capable of mediating a persistent depolarizing drive to the soma and which probably mediate the bistable behaviour of these cells. PMID- 10792442 TI - N-methyl-D-aspartate enhancement of the glycine response in the rat sacral dorsal commissural neurons. AB - The effect of N-methyl-D-aspartate (NMDA) on the glycine (Gly) response was examined in neurons acutely dissociated from the rat sacral dorsal commissural nucleus (SDCN) using the nystatin-perforated patch-recording configuration under voltage-clamp conditions. The application of 100 microM NMDA to SDCN neurons reversibly potentiated Gly-activated Cl- currents (IGly) without affecting the Gly binding affinity and the reversal potential of IGly. A selective NMDA receptor antagonist, APV (100 microM), blocked the NMDA-induced potentiation of IGly, whereas 50 microM CNQX, a non-NMDA receptor antagonist, did not. The potentiation effect was reduced when NMDA was applied in a Ca2+-free extracellular solution or in the presence of BAPTA AM, and was independent of the activation of voltage-dependent Ca2+ channels. Pretreatment with KN-62, a selective Ca2+-calmodulin-dependent protein kinase II (CaMKII) inhibitor, abolished the NMDA action. Inhibition of calcineurin (CaN) further enhanced the NMDA-induced potentiation of IGly. In addition, the GABAA receptor-mediated currents were suppressed by NMDA receptor activation in the SDCN neurons. The present results show that Ca2+ entry through NMDA receptors modulates the Gly receptor function via coactivation of CaMKII and CaN in the rat SDCN neurons. This interaction may represent one of the important regulatory mechanisms of spinal nociception. The results also suggest that GABAA and Gly receptors may be subject to different intracellular modulatory pathways. PMID- 10792443 TI - Cross-modal plasticity of the corticothalamic circuits in rats enucleated on the first postnatal day. AB - Reorganization of the reciprocal corticothalamic connections was studied as a possible anatomical substrate of the cross-modal compensation of the missing visual input of the visual cortex by somatosensory-evoked activities in neonatally enucleated rats. The use of quantitative retrograde tract-tracing techniques revealed that the contribution of the lateral posterior thalamic nucleus (LP) is significantly increased following enucleation, while that of the dorsolateral geniculate and the lateral dorsal nuclei is decreased in the thalamocortical afferentation of a region in visual cortical area 17. In contrast with the control rats, a dense terminal arborization of afferents was labelled in the LP after the injection of anterograde tracer into the barrel cortex of the enucleated rats. The injection of anterograde tracer into the visual cortex also demonstrated a massive afferentation into the LP of the enucleated rats. Visual and somatosensory corticothalamic afferents exhibited similar ultrastructural features in the LP after enucleation, but their synaptic organizations differed as regards the diameter of the postsynaptic dendrites. Taken together with the previous observations, these results suggest a central role for the LP in the transmission of the somatosensory-evoked activities to the visual cortex after early blindness. PMID- 10792444 TI - Expression of novel antioxidant thioredoxin-2 in the rat brain. AB - Thioredoxins are a class of small redox-regulating proteins that have been implicated in the control of various aspects of cellular functions and seem to be one of the key regulators of signalling in the cellular responses to various stresses. Thioredoxin-2 (Trx2) is a novel mammalian thioredoxin which, in contrast to previously known cytosolic thioredoxin (Trx1), has been localized to the mitochondria. Trx2 is abundantly expressed in skeletal muscle, heart and adrenal gland, as well as in some other peripheral tissues with high metabolic activity. Using in situ hybridization and immunohistochemistry, we have studied the distribution and regulation of Trx2 expression in the rat brain. Trx2 mRNA and protein are highly expressed in the neurons in several brain regions, including the olfactory bulb, frontal cortex, hippocampus, some hypothalamic and thalamic nuclei, cerebellum and numerous brainstem nuclei. In addition, the Trx2 mRNA expression in paraventricular hypothalamic nucleus and reticular thalamic nucleus was found to be sensitive to peripheral glucocorticoids, as dexamethasone treatment caused significant elevation of Trx2 mRNA level in this area. No changes in other brain areas were observed after dexamethasone treatment. These findings implicate a significant regulatory and/or protective function of Trx2 in the nervous system. PMID- 10792445 TI - Participation of low-threshold calcium spikes in excitatory synaptic transmission in guinea pig medial frontal cortex. AB - We studied the activation of low-threshold calcium spikes (LTS) by excitatory postsynaptic potentials in pyramidal neurons from guinea pig medial frontal cortex with intracellular recording. We used extracellular bicuculline and phaclofen and intracellular QX-314 to block inhibitory synaptic potentials and sodium currents. Postsynaptic potentials were evoked by stimulation of layer I. We found that large (> 10-15 mV) excitatory synaptic potentials evoked from membrane potentials more negative than -75 mV were able to trigger LTS. The activation of LTS resulted in an increase of the rising slope or amplitude of the synaptic potentials depending on the size of the excitatory postsynaptic potential (EPSP). We used 100 microM NiCl2 to confirm the presence of LTS as part of the EPSPs. The N-methyl-D-aspartate (NMDA) and non-NMDA components of the excitatory synaptic potentials were isolated using (+/-)2-amino-5 phosphonovaleric acid (APV; 50 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM); both components could, independently, trigger an LTS. With recordings made with K+ acetate-filled electrodes, we show that the activation of LTS was critical to allow excitatory synaptic potentials to reach the threshold of action potential firing; also, this amplification of synaptic responses produced the firing of more than a single action potential by the postsynaptic cell. These results demonstrate that in cortical pyramidal neurons the activation of low-threshold calcium spikes results in the amplification of synaptic responses. PMID- 10792446 TI - Seizures induce widespread upregulation of cystatin B, the gene mutated in progressive myoclonus epilepsy, in rat forebrain neurons. AB - Loss of function mutations in the gene encoding the cysteine protease inhibitor, cystatin B (CSTB), are responsible for the primary defect in human progressive myoclonus epilepsy (EPM1). CSTB inhibits the cathepsins B, H, L and S by tight reversible binding, but little is known regarding its localization and physiological function in the brain and the relation between the depletion of the CSTB protein and the clinical symptoms in EPM1. We have analysed the expression of mRNA and protein for CSTB in the adult rat brain using in situ hybridization and immunocytochemistry. In the control brains, the CSTB gene was differentially expressed with the highest levels in the hippocampal formation and reticular thalamic nucleus, and moderate levels in amygdala, thalamus, hypothalamus and cortical areas. Detectable levels of CSTB were found in virtually all forebrain neurons but not in glial cells. Following 40 rapidly recurring seizures evoked by hippocampal kindling stimulations, CSTB mRNA expression showed marked bilateral increases in the dentate granule cell layer, CA1 and CA4 pyramidal layers, amygdala, and piriform and parietal cortices. Maximum levels were detected at 6 or 24 h, and expression had reached control values at 1 week post-seizures. The changes of mRNA expression were accompanied by transient elevations (at 6-24 h) of CSTB protein in the same brain areas. These findings demonstrate that seizure activity leads to rapid and widespread increases of the synthesis of CSTB in forebrain neurons. We propose that the upregulation of CSTB following seizures may counteract apoptosis by binding cysteine proteases. PMID- 10792447 TI - Transplanted CNS stem cells form functional synapses in vivo. AB - An understanding of developmental mechanisms and new cell therapies can be achieved by transplantation into the nervous system. Multipotential stem cells have been isolated from the foetal and adult central nervous system (CNS). Immortalized and primary precursor cells integrate into the developing brain generating both neurons and glia as defined by immunological and morphological criteria. Here we show for the first time that in vitro-expanded CNS precursors, upon transplantation into the brains of rats, form electrically active and functionally connected neurons. These neurons exhibit spontaneous and evoked postsynaptic events and respond to focal glutamate application. Donor cells were grafted into the foetal hippocampus, and the amplitude and frequency of spontaneous synaptic events were monitored in the grafted cells in area CA1 for the first month of postnatal life. The formation of synapses onto grafted neurons indicates that grafted CNS stem cells can be used to study synaptic development in vivo and has important implications for clinical cell replacement therapies. PMID- 10792449 TI - Disconnecting hippocampal projections to the anterior thalamus produces deficits on tests of spatial memory in rats. AB - A disconnection procedure was used to test whether projections from the hippocampus to the anterior thalamic nuclei (AT), via the fimbria-fornix (FX), form functional components of a spatial memory system. The behavioural effects of combined unilateral lesions in the AT and FX were compared when they were either in contralateral hemispheres (AT-FX Contra) or the same hemisphere (AT-FX Ipsi). Other groups received bilateral FX lesions and Sham surgeries. Expt 1 demonstrated that none of these lesions affected performance of an object recognition task, while performance of an object location task, which tests the subjects' preference for an object that has changed location, was impaired in the AT-FX Contra and FX groups. In a T-maze alternation task, however, the FX group was severely impaired while both the AT-FX Ipsi and AT-FX Contra lesion groups showed only a mild impairment. In order to test whether spared crossed projections might support spatial performance in the AT-FX Contra group we then examined the effects of a combined AT-FX Contra lesion coupled with transection of the hippocampal commissure. This combination of lesions produced a severe disruption in spatial memory performance in the water maze, radial arm maze and T maze, which was significantly greater than that produced by ipsilateral and contralateral AT-FX lesions alone. These results support the notion that disconnection of the AT from their hippocampal inputs produces impairments on a range of spatial memory tasks, but indicate that there are an array of different routes that can subserve this function. PMID- 10792448 TI - Involvement of the medial septum in stress-induced relapse to heroin seeking in rats. AB - Intermittent footshock stress has been shown to reinstate extinguished drug taking behaviour in rats, but the brain areas involved in this effect are to a large degree unknown. Here we studied the role of the septum in stress-induced reinstatement of heroin seeking. Rats were trained to self-administer heroin for 9-10 days (three 3-h sessions per day, 0.1 mg/kg per infusion). Following training, extinction sessions were given for 8-13 days by substituting saline for heroin, and then tests for reinstatement of heroin seeking were carried out. Reversible inactivation of the medial septum with tetrodotoxin (TTX; 1-5 ng, infused 25-40 min before the test sessions) reliably reinstated heroin seeking, mimicking the effect of 15 min of intermittent footshock. This effect of TTX was not observed after infusions made 1.5 mm dorsally into the lateral septum. In other experiments, it was found that infusions of a low, subthreshold dose of TTX (0.5 ng) into the medial septum, when combined with 2 min of footshock that in itself was ineffective, reinstated heroin seeking. Furthermore, electrical stimulation (400 microA pulses, 100 micros duration, 100 Hz frequency) of the medial septum during exposure to 10 min of intermittent footshock attenuated footshock-induced reinstatement of heroin seeking. These data suggest a role for the medial septum in stress-induced relapse to drug seeking. The septum is thought to be involved in neuronal processes underlying behavioural inhibition, thus we speculate that stressors provoke relapse by interfering with these processes. PMID- 10792450 TI - Cellular response to texture and form defined by motion in area 19 of the cat. AB - The present study examined the neuronal sensitivity in area 19 of the cat to a motion-defined bar and to texture. Sensitivity was tested in normal, lesioned (areas 17-18) and split-chiasm cats using a kinematogram, as well as a textured bar drifting on a uniform light background and a light bar drifting on a stationary textured background. Texture density was varied. The results indicate that almost all cells of area 19 recorded in the three groups of cats responded to a motion-defined bar or to its edges. Texture density influenced the responses in that the discharge rate increased as density decreased. However, the majority of cells were sensitive to the highest texture density kinematogram. Moreover, the neural responses of all cats were either independent of the density of the textured bar or background, or were modulated by it. These results show that cells in area 19 can signal the presence of a kinetic bar and that the density of either the textured bar, the background or both can influence figure-ground detection. The results are interpreted with respect to how various inputs influence the function of area 19. PMID- 10792451 TI - Coding for auditory space in the superior colliculus of the rat. AB - Although the rat is often used to determine behavioural sound-localization capabilities or neuronal computation of binaural information, the representation of auditory space in the rat brain has not been investigated so far. We obtained extracellular recordings from auditory neurons in the superior colliculus of anaesthetized rats and examined them for spatial tuning characteristics and topographical order. Many neurons (73%) showed significant tuning, with a single peak in the azimuth response profiles based on spike rates and response latencies. Best azimuth values from neurons in one SC were generally tuned to contralateral and rarely to frontal or ipsilateral directions. Tuning width was mostly broad; at supra-threshold sound pressure levels (35 dB SPL), 55% of the units had a tuning width of > 120 degrees in contralateral space. Additionally, tuning width increased with stimulation intensity. A significant but considerably scattered topographical order of best azimuth directions was observed in the deep layers of the superior colliculus with frontal directions being represented closer to the rostral pole. Tuned auditory units in the intermediate layers of the superior colliculus, however, showed no systematic spatial arrangement. This pattern was confirmed by analysing best azimuth directions from simultaneously recorded units. Our results indicate that the rat superior colliculus contains a representation of auditory space which is similar to that described for other small mammals. PMID- 10792452 TI - Neural selectivity for hue and saturation of colour in the primary visual cortex of the monkey. AB - In the inferior temporal (IT) cortex of monkeys, which has been shown to play a critical role in colour discrimination, there are neurons sensitive to a narrow range of hues and saturation. By contrast, neurons in the retina and the parvocellular layer of the lateral geniculate nucleus (pLGN) encode colours in a way that does not provide explicit representation of hue or saturation, and the process by which hue- and saturation-selectivity is elaborated remains unknown. We therefore tested the colour-selectivity of neurons in the primary visual cortex (V1) and compared it with those of pLGN and IT neurons. Quantitative analysis was performed using a standard set of colours, systematically distributed within the CIE (Commission Internationale de l'Eclairage)-xy chromaticity diagram. Selectivity for hue and saturation was characterized by analysing response contours reflecting the overall distribution of responses across the chromaticity diagram. We found that the response contours of almost all pLGN neurons were linear and broadly tuned for hue. Many V1 neurons behaved similarly; nonetheless, a considerable number of V1 neurons had clearly curved response contours and were selective for a narrow range of hues or saturation. The relative frequencies of neurons exhibiting various selectivities for hue and saturation were remarkably similar in the V1 and IT cortex, but were clearly different in the pLGN. Thus, V1 apparently plays a very important role in the conversion of colour signals necessary for generating the elaborate colour selectivity observed in the IT cortex. PMID- 10792453 TI - Differential amygdala responses to winning and losing: a functional magnetic resonance imaging study in humans. AB - The amygdala has been shown to respond to many distinct types of affective stimuli, including reward and punishment feedback in animals. In humans, winning and losing situations can be considered as reward and punishment experiences, respectively. In this study, we used functional magnetic resonance imaging (fMRI) to measure regional brain activity when human subjects were given feedback on their performance during a simple response time task in a fictitious competitive tournament. Lexical stimuli were used to convey positive 'win' or negative 'lose' feedback. The frequency of positive and negative trials was parametrically varied by the experimenters independently from the subjects' actual performance and unbeknownst to them. The results showed that the parametric increase of winning was associated with left amygdala activation whereas the parametric increase of losing was associated with right amygdala activation. These findings provide functional evidence that the human amygdala differentially responds to changes in magnitude of positive or negative reinforcement conveyed by lexical stimuli. PMID- 10792454 TI - Protection against dopaminergic nigrostriatal cell death by excitatory input ablation. AB - The importance of enhanced glutamatergic neurotransmission in the basal ganglia and related structures has recently been highlighted in the development of Parkinson's disease. The pedunculopontine tegmental nucleus (PPN) is the major origin of excitatory, glutamatergic input to dopaminergic nigrostriatal neurons of which degeneration is well known to cause Parkinson's disease. Based on the concept that an excitatory mechanism mediated by glutamatergic neurotransmission underlies the pathogenesis of neurodegenerative disorders, we made an attempt to test the hypothesis that removal of the glutamatergic input to the nigrostriatal neurons by PPN lesions might prevent 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism in the macaque monkey. The PPN was lesioned unilaterally with microinjection of kainic acid, and, then, MPTP was administered systemically. In these monkeys, the degree of parkinsonian motor signs was behaviourally evaluated, and the histological changes in the dopaminergic nigrostriatal system were analysed by means of tyrosine hydroxylase immunohistochemistry. The present results revealed that nigrostriatal cell loss and parkinsonian motor deficits were largely attenuated in the MPTP-treated monkey group whose PPN had been lesioned, compared with the control, MPTP-treated monkey group with the PPN intact. This clearly indicates that the onset of MPTP neurotoxicity is suppressed or delayed by experimental ablation of the glutamatergic input to the nigrostriatal neurons. Such a protective action of excitatory input ablation against nigrostriatal cell death defines evidence that nigral excitation driven by the PPN may be implicated in the pathophysiology of Parkinson's disease. PMID- 10792455 TI - Effects of scopolamine on a novel choice serial reaction time task. AB - Rats were trained on a novel attentional task adapted from the five-choice serial reaction time test first developed by Carli et al. [1983; Behav. Brain. Res., 9, 361-80]. The novel task required rats to detect the occurrence of brief light flashes in one of two spatial locations following trial initiation by a lever press. Blank trials were interleaved with stimulus trials and the rat had to make a different response to indicate the absence of a light. Occasional light-alone trials were also presented in which the visual stimulus appeared without forewarning. Pre-exposure to food for 60 min prior to test increased response omissions for all trials and slowed correct response latency, but failed to significantly alter accuracy. A decrease in light-stimulus duration (1-0.4 s) decreased accuracy, increased the reporting of blank trials and the number of light-alone trial omissions. Scopolamine (0.03-0.1 mg/kg) and scopolamine methylbromide (0.1 mg/kg) failed to affect accuracy, but increased light-alone trial omissions and lengthened correct response latency. The results confirm that the novel task is able to distinguish between motivational and attentional manipulations, and imply that scopolamine affected performance via nonattentional mechanisms. PMID- 10792456 TI - Dopamine-independent action of cocaine on striatal and accumbal neurons. AB - Increasing evidence suggests that dopamine (DA) mechanisms alone cannot fully explain the psychoemotional and behavioural effects of cocaine, including its ability to induce drug-taking behaviour. Although it is known that cocaine, after intravenous administration or smoking, may reach brain levels high enough to inhibit Na+ transport, the role of this action remains unclear. To examine the contribution of local anaesthetic and DA mechanisms to changes in striatal and accumbal neuronal activity induced by cocaine, single-unit recording was combined with iontophoresis in awake, unrestrained rats. Most spontaneously active and glutamate-stimulated neurons were highly sensitive to brief cocaine applications (0-40 nA); cocaine-induced inhibitions occurred at small ejection currents (0-5 nA), were dose-dependent, highly stable during repeated applications and strongly dependent on basal activity rates. These neuronal responses remained almost unchanged after systemic administration of either a selective D1 antagonist (SCH 23390, 0.2 mg/kg) or a combination of SCH-23390 (1 mg/kg) and eticlopride (1 mg/kg), a D2 antagonist. Whereas SCH-23390 alone had a weak attenuating effect, no effect and even a slight enhancement of responses to cocaine occurred in fast firing glutamate (GLU)-stimulated units after the combined blockade of D1 and D2 receptors. Responses to cocaine were mimicked by iontophoretic procaine (0-40 nA), a short-acting local anaesthetic with minimal effect on DA uptake. Procaine induced inhibitions occurred at the same low currents, had a similar time-course, and were also strongly dependent on basal discharge rate. Our data support the existence of a DA-independent mechanism for the action of cocaine involving a direct interaction with Na+ channels. Although further studies are required to clarify this mechanism and its interaction with other pharmacological and behavioural variables, a direct interaction with Na+ channels may contribute to changes in neuronal activity induced by self-injected cocaine, thereby playing a role in mediating the psychoemotional and behavioural effects of this drug. PMID- 10792457 TI - Influence of the predicted time of stimuli eliciting movements on responses of tonically active neurons in the monkey striatum. AB - Changes in activity of tonically active neurons of the primate striatum are determined both by the behavioural significance of stimuli and the context in which stimuli are presented. We investigated how the responses of these neurons are modified by the temporal predictability of stimuli eliciting learned behavioural reactions. Single neurons were recorded from the caudate nucleus and putamen of two macaque monkeys performing a visual reaction time task under conditions in which the timing of the trigger stimulus was made more or less predictable. The monkeys' ability to predict the trigger onset was assessed by measuring arm movement reaction times and saccadic ocular reactions. Of 171 neurons responding to the unsignalled presentation of the trigger stimulus, 32% lost their response when an instruction cue preceded the trigger by a highly practised 1.5 s interval, and the response reappeared when this interval was varied randomly from 1 to 2.5 s or prolonged to 3 or 4. 5 s. Although 43% of the neurons remained responsive irrespective of task condition, the responses were stronger with longer intervals than with the accustomed 1.5 s interval. In addition, a number of neurons responding to the instruction lost their response when the trigger appeared more distant from the instruction. These findings demonstrate that neuronal responses to a movement-triggering signal become more numerous and pronounced when the degree of temporal predictability of that signal was decreased. We conclude that tonic striatal neurons are sensitive to temporal aspects of stimulus prediction. PMID- 10792458 TI - X-ray kinematic analysis of forelimb movements during target reaching and food taking in the cat. AB - We have used a three-dimensional X-ray cinematographic approach to investigate the kinematics of the forelimb during target reaching and food taking in five cats. Measurements of the trajectory of the limb during the reaching movement showed that the movement paths of the metacarpophalangeal joint (MCP) and the wrist were sigmoidal with a long nearly linear segment. The elbow followed a bent movement path with maximal inflection in the middle. The path of the humerus had an ascending parabola-like characteristic. The velocity profiles of the MCP and wrist were nearly bell-shaped and skewed to the left, whereas the profiles of the elbow joint were more or less double peaked with the second peak occurring 60-40 ms before object contact. Several different velocity peaks reflecting specific aspects of the task existed when the bell-shaped velocity profiles were divided in their vectorial components. Angular motion of the elbow consisted of a flexion extension sequence during the reach and a flexion during the subsequent retraction. After an initial flexion during lift-off the wrist was extended. It kept this extended position during orienting towards the food container. During the retraction phase it was further extended. The angle between the wrist axis and the parasagittal plane changed during the movement. It first increased, then decreased during the last 100 ms before the object was reached. During the retraction it increased again to support the object weight against gravity. The position of the wrist was established by radio-ulnar supination and movements of the whole arm around the shoulder joint. We hypothesize that the position of the wrist axis is the controlled variable during protraction and retraction, regardless of whether it is achieved by radio-ulnar supination or by movements around the shoulder. PMID- 10792459 TI - Increased rewarding properties of morphine in dopamine-transporter knockout mice. AB - The activation of dopamine (DA) neurotransmission plays a crucial role in the behavioural responses to drugs of abuse. In particular, increased extracellular levels of DA within the mesolimbic pathway have been implicated in the rewarding and locomotor stimulatory properties of morphine. We investigated the behavioural responses to morphine in mice with a genetic disruption of the DA transporter (DAT), resulting in a constitutively high level of extrasynaptic DA. In the conditioned place preference test, DAT-/- mice exhibited a stronger rewarding response to morphine (5 mg/kg, s.c.) compared with control littermates. However, the same dose of morphine failed to increase locomotor activity in DAT-/- mice, whilst enhancing locomotion in DAT+/- and DAT+/+ animals. Morphine-induced analgesia was unaffected in mutant mice, but the behavioural expression of naloxone-induced withdrawal signs was blunted. In vivo voltammetry in the shell of the nucleus accumbens revealed that morphine was able to stimulate DA neurons in DAT-/- mice, resulting in the accumulation of higher extracellular DA levels compared with control animals. Morphine also induced a higher rate of c-fos transcription in the shell of the nucleus accumbens in mutant mice. We conclude that morphine-induced rewarding responses are firmly established in DAT mutant mice despite a DA transmission that is already tonically activated, and independently of any effect on locomotion. These particular behavioural responses to morphine may be associated with the action of the drug on DA release and c-fos expression in the shell of the nucleus accumbens of DAT-/- mice. PMID- 10792460 TI - Depolarization-dependent survival of cultured mouse cerebellar granule neurons is strain-restrained. AB - Cerebellar granule neurons (CGNs) isolated from the rat are often used as a model system for the analysis of activity-dependent survival of neurons. These cells do not survive in culture without addition of a depolarizing agent (KCl or glutamate) to the medium. However, it has been reported that mouse CGN behave differently. Here we found that the requirement for depolarization for the survival of the mouse CGN was strain-dependent, which may be important for future analyses using transgenic animals. CGNs from the Balb/C mouse could survive without KCl addition, whereas CGNs from the C57Bl/6 mouse could not (similar to the rat CGN). The survival-promoting activity of the Balb/C mouse CGNs was transferable by coculturing. However, the medium of the Balb/C mouse CGN culture was ineffective, suggesting that the neurotrophic substance that is supposed to be released is labile. Although we are yet to specify the substance, it might be independent of neurotrophins. It is necessary to select the strain of mouse in the production of transgenic animals for the analyses of activity-dependent neuronal survival. PMID- 10792461 TI - Modulation of synaptic transmission in the rat ventral septal area by the pharmacological activation of metabotropic glutamate receptors. AB - The ventral septal area (VSA) is considered to be critically involved in the control of the height and duration of fever. The major excitatory input to this region of the brain is glutamatergic, and the aim of this study was to investigate possible modulation of this synapse by metabotropic glutamate (mGlu) receptors. Whole-cell patch recordings were made from individual VSA neurons voltage-clamped at -60 mV. Activation of either group I or group II mGlu receptors (by bath application of 3,5-dihydroxyphenylglycine (DHPG) or (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV), respectively) produced a long-lasting depression of synaptic transmission which in both cases was insensitive to the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino 5-phosphonopentanoate (D-AP5). In contrast, application of (S)-2-amino-4 phosphonobutyric acid (L-AP4), a group III mGlu receptor agonist, had a biphasic effect on synaptic transmission in the VSA, first eliciting a transient depression of transmission during drug application, followed by a marked and sustained potentiation of synaptic transmission upon drug washout. The response elicited by L-AP4 was dependent on NMDA receptor activation, as in the presence of D-AP5 the potentiation was replaced by an underlying long-term depression (LTD) of transmission. These data provide the first evidence that metabotropic glutamate receptor agonists can induce both NMDA receptor-dependent and independent modulation of synaptic transmission in the VSA. PMID- 10792462 TI - Increased responsivity of glutamate release from the substantia nigra pars reticulata to striatal NMDA receptor blockade in a model of Parkinson's disease. A dual probe microdialysis study in hemiparkinsonian rats. AB - Dual probe microdialysis was employed in freely moving 6-hydroxydopamine (6-OHDA) hemilesioned rats to investigate the effects of blockade of N-methyl-D-aspartate (NMDA) receptors in the dorsolateral striatum on glutamate (Glu) release from the ipsilateral substantia nigra pars reticulata (SNr). Perfusion for 60 min with the NMDA antagonist dizocilpine (0.1 and 1 microM) in the dopamine (DA)-denervated striatum stimulated nigral Glu release (peak effect of 139 +/- 7% and 138 +/- 9%, respectively). The lower (0.01 microM) and higher (10 microM) concentrations were ineffective. In sham-operated rats, dizocilpine failed to affect nigral Glu release up to 1 microM but induced a prolonged stimulation at 10 microM (153 +/- 9% at the end of perfusion). The present results show that DA-deficiency in the striatum of hemiparkinsonian rats is associated with increased responsivity of nigral Glu release to striatal NMDA receptor blockade. This suggests that changes of NMDA receptor mediated control of the striatofugal pathways occur during Parkinson's disease (PD). PMID- 10792463 TI - Mechanism of catabolite repression in the bgl operon of Escherichia coli: involvement of the anti-terminator BglG, CRP-cAMP and EIIAGlc in mediating glucose effect downstream of transcription initiation. AB - BACKGROUND: Expression of the bgl operon of Escherichia coli, involved in the regulated uptake and utilization of aromatic beta-glucosides, is extremely sensitive to the presence of glucose in the growth medium. We have analysed the mechanism by which glucose exerts its inhibitory effect on bgl expression. RESULTS: Our studies show that initiation of transcription from the bgl promoter is only marginally sensitive to glucose. Instead, glucose exerts a more significant inhibition on the elongation of transcription beyond the rho independent terminator present within the leader sequence. Transcriptional analyses using plasmids that carry mutations in bglG or within the terminator, suggest that the target for glucose-mediated repression is the anti-terminator protein, BglG. Introduction of multiple copies of bglG or the presence of mutations that inhibit its phosphorylation by Enzyme IIBgl (BglF), result in loss of glucose repression. Studies using crp, cya and crr strains show that both CRP cAMP and the Enzyme IIAGlc (EIIAGlc) are involved in the regulation. Although transcription initiation is normal in a crp, cya double mutant, no detectable transcription is seen downstream of the terminator, which is restored by a mutation within the terminator. Transcription past the terminator is also partly restored by the addition of exogenous cAMP to glucose-grown cultures of a crp+ strain. Glucose repression is lost in the crr mutant strain. CONCLUSIONS: The results summarized above indicate that glucose repression in the bgl operon is mediated at the level of transcription anti-termination, and glucose affects the activity of BglG by altering its phosphorylation by BglF. The CRP-cAMP complex is also involved in this regulation. The results using the crr mutant suggest a negative role for EIIAGlc in the catabolite repression of the bgl genes. PMID- 10792464 TI - Functional interaction of general transcription initiation factor TFIIE with general chromatin factor SPT16/CDC68. AB - BACKGROUND: Transcriptional initiation of class II genes is one of the major targets for the regulation of gene expression and is carried out by RNA polymerase II and many auxiliary factors, which include general transcription initiation factors (GTFs). TFIIE, one of the GTFs, functions at the later stage of transcription initiation. As recent studies indicated the possibility that TFIIE may have a role in chromatin transcriptional regulation, we isolated TFIIE interacting factors which have chromatin-related functions. RESULTS: Using the yeast two-hybrid screening system, we isolated the C-terminal part of the human homologue of Saccharomyces cerevisiae (y) Spt16p/Cdc68p, a general chromatin factor. The C-terminal part of human SPT16/CDC68 directly interacts with TFIIE, and ySpt16p/Cdc68p also interacts with yTFIIE (Tfa1p/Tfa2p), thus indicating the existence of an evolutionarily conserved interaction between TFIIE and SPT16/CDC68. Functional interaction of yTFIIE and ySpt16p/Cdc68p was examined using a conditional yTFIIE-alpha mutant strain. Over-expression of ySpt16p/Cdc68p suppressed the phenotype of cold sensitivity of the yTFIIE-alpha-cs mutant strain, and in vitro binding assays revealed that yTFIIE-alpha-cs mutant protein showed diminished binding affinity to ySpt16p/Cdc68p. CONCLUSIONS: These observations indicate that general transcription initiation factor TFIIE functionally interacts with general chromatin factor SPT16/CDC68, a finding which provides new insight into the involvement of TFIIE in chromatin transcription. This may well lead to a breakthrough in relationships between the transcription initiation process and structural changes in chromatin. PMID- 10792465 TI - Identification and characterization of testis specific ornithine decarboxylase antizyme (OAZ-t) gene: expression in haploid germ cells and polyamine-induced frameshifting. AB - BACKGROUND: Polyamines are known to play important roles in the proliferation and differentiation of many types of cells. However, in the testis, where polyamines such as spermidine and spermine exist in high concentrations, their roles still remains to be elucidated. RESULTS: We have cloned a testis-specific gene encoding an ornithine decarboxylase antizyme known to control intracellular concentrations of polyamines in a feedback manner. The mRNA encoding the protein named ornithine decarboxylase antizyme in testis (OAZ-t) was specifically expressed in haploid germ cells. In contrast, the mRNA level of somatic ornithine decarboxylase antizyme 1 (OAZ1) decreased markedly at the late stages of haploid germ cell differentiation. OAZ-t mRNA was first observed in 23-day-old mice, whereas the OAZ-t protein was detected much later, at 35 days after birth. Further experiments on OAZ-t revealed that polyamines were capable of inducing a frameshifting at the frameshift sequence of OAZ-t mRNA, resulting in the translation of OAZ-t, as was the case with the somatic OAZ1. Transfection of OAZ t cDNA inactivated the ornithine decarboxylase activity in the HEK293 cells. CONCLUSIONS: Results indicate that the expression of OAZ-t is controlled at both transcriptional and translational levels, and that OAZ-t likely plays a key role in spermatogenesis by regulating the intracellular concentration of polyamines in haploid germ cells. PMID- 10792466 TI - Intronic U50 small-nucleolar-RNA (snoRNA) host gene of no protein-coding potential is mapped at the chromosome breakpoint t(3;6)(q27;q15) of human B-cell lymphoma. AB - BACKGROUND: In eukaryotic cells, nucleolar processing of preribosomal RNAs (prerRNAs) is assisted by a large number of small nucleolar RNAs (snoRNAs) that function in the 2'-O-methylation or the pseudouridylation of rRNAs. Most snoRNAs so far characterized are encoded and processed from introns of premRNAs. RESULTS: We found a novel intronic snoRNA gene, named U50HG, located on chromosome 6q15, at the breakpoint of chromosomal translocation t(3;6)(q27;q15). The U50HG gene is composed of six exons, whose spliced transcripts have little potential for coding a protein, and its introns produce both U50 and U50-like (U50') snoRNAs that are localized in nucleoli. It possesses an oligopyrimidine tract that is characteristic of the 5'-terminal oligopyrimidine (5'TOP) class of genes which have been shown to be coordinately regulated in response to cell growth. CONCLUSIONS: U50HG is a member of the nonprotein-coding multiple snoRNA host gene family, as well as of the 5'TOP gene family similar to UHG (U22 host gene), U17HG (U17 host gene), U19HG (U19 host gene) and gas5 (growth arrest-specific transcript 5 gene). It is novel to find that the snoRNA gene is located at the breakpoint of chromosomal translocation t(3;6)(q27;q15) involved in human B-cell lymphoma. PMID- 10792467 TI - Rev-dependent association of the intron-containing HIV-1 gag mRNA with the nuclear actin bundles and the inhibition of its nucleocytoplasmic transport by latrunculin-B. AB - BACKGROUND: A hallmark of HIV-1 gene expression is that unspliced genomic RNA, which also acts as mRNA for the expression of Gag/Pol, is exported to the cytoplasm. Rev directs this transport through the nuclear export signal (NES). RESULTS: Fluorescence in situ hybridization and immunocytochemistry demonstrated that gag mRNA, Rev, and its NES receptor, CRM1, and RanGTPase formed nuclear tracks which were congruent with underlying beta-actin bundles. Actin bundle formation was confirmed electron-microscopically. These bundles were observed upon Rev-containing gag RNP formation. The loss of bundles was associated with the nuclear retention of gag mRNA. Reverse transcription-polymerase chain reaction analysis of both cytoplasmic and nuclear gag mRNAs demonstrated that disruption of nuclear actin filament formation by latrunculin-B (LAT-B), an F actin depolymerizing compound, resulted in the dose-dependent inhibition of gag mRNA export. The differential subtyping of the mRNA-positive cells confirmed morphologically the effect of LAT-B treatment. The export inhibition was specific to gag mRNA and export of fully spliced HIV-1 tat/rev mRNAs as well as cellular GAPDH mRNA was not affected by the compound. CONCLUSIONS: Nuclear beta-actin bundles are suggested to be functionally involved in the Rev-dependent nucleocytoplasmic transport of intron-containing HIV-1 gag mRNA. PMID- 10792468 TI - Blockade of cadherin-6B activity perturbs the distribution of PSD-95 family proteins in retinal neurones. AB - BACKGROUND: Synaptic junctions have cadherin-catenin complexes, but their functions are poorly understood. Using retinal neurones, we investigated the role of this adhesion machinery in synaptic organization. RESULTS: In cultures of chicken retinal cells, cadherin-6B (cad6B) and cadherin-7 (cad7) are expressed by distinct neurones, each being distributed in a punctate pattern along their neurites as well as in the soma. Double-immunostaining for cad6B and PSD-95/SAP90 or other PSD-95 family members, known to localize in the postsynaptic density, showed that their distributions overlapped each other. To assess the role for cad6B, we incubated retinal cells with antibodies that could specifically block cad6B-mediated adhesion. In the antibody-treated neurones, the localization pattern of PSD-95 family proteins was altered, that is, their staining signals tended to be reduced or disarranged. We then examined whether cadherins interacted molecularly with PSD-95: Cadherin immunoprecipitates from brain lysates did not contain PSD-95; nevertheless, this protein was co-precipitated with alphaN- and beta-catenins. When PSD-95 proteins were ectopically expressed in epithelial cells, some of these molecules were concentrated in cell-cell junctions, co-localizing with E-cadherin, and this junctional localization of PSD 95 was abolished by blocking of E-cadherin activity. CONCLUSION: These results suggest that cadherins play a role in the subcellular organization of postsynaptic density components through some, perhaps indirect, molecular interactions. PMID- 10792469 TI - Establishing haemophilia care in developing countries: using data to overcome the barrier of pessimism. PMID- 10792470 TI - Influence of factor VIII:C and factor IX activity in plasmas of haemophilic dogs on the activated partial thromboplastin time measured with two commercial reagents. AB - The present study is based on 145 plasma samples with a reduced activity of factor VIII:C (range: 0.009-0.62 IU mL-1) and 28 samples with a reduced factor IX activity (range: 0.035-0.55 IU mL-1). The samples were collected from dogs with haemophilia A (n=22) or haemophilia B (n=3), some of these during substitution therapy. For all samples the activated partial thromboplastin time (APTT) was measured with two commercial reagents containing kaolin as a contact activator. In each case, the deficiency of factor VIII:C or IX was reflected in abnormal results of the APTT. This was true for both reagents. A significant correlation (P < 0.001) was found between factor VIII:C activity and APTT (reagent 1, Pathromtin(R); Spearman's rank correlation coefficient, rS=-0.731, reagent 2, PTT Reagenz; rS=-0.875) as well as between factor IX activity and APTT (reagent 1, rS=-0.819; reagent 2, rS=-0.955]. In each case, the relationship between coagulation factor activity and APTT could be proven most precisely by geometric regression. The results of this study illustrate the applicability of commercial APTT test kits as a sensitive screening test of factor VIII:C and IX deficiencies in canine plasma. PMID- 10792471 TI - Safety and efficacy of solvent/detergent-treated antihaemophilic factor with an added 80 degrees C terminal dry heat treatment in patients with haemophilia A. AB - Plasma-derived factor VIII concentrates remain an important resource for haemophilia A patients. To improve the safety of these preparations, various methods of viral removal and inactivation have been used that are designed to eliminate both enveloped and non-enveloped viruses. There have been rare reports that some viral inactivation processes altered the immunogenicity of some concentrates, leading to the development of factor VIII inhibitors in previously treated haemophilia A patients. This study evaluated the safety, efficacy and lack of neo-antigenicity of a highly purified factor VIII preparation which undergoes both solvent/detergent treatment and final dry heat treatment at 80 degrees C for 72 h. The study included: (i) a single blind, single-dose crossover pharmacokinetic study in 18 previously treated patients, comparing sibling lots of the unheated preparation (Koate(R)-HP) and the heat-treated preparation (Koate(R)-DVI), and (ii) an extended home treatment programme for 36 patients at two haemophilia treatment centres primarily to assess immunogenicity. Clinical parameters were assessed at regular intervals. The results confirm that Koate(R) HP and Koate(R)-DVI are bioequivalent, and that Koate(R)-DVI is safe and efficacious for treatment of acute bleeding episodes and for surgery. Furthermore, the heat-treated preparation is not associated with the development of inhibitors in previously treated patients. PMID- 10792472 TI - Immune tolerance induction in the treatment of paediatric haemophilia A patients with factor VIII inhibitors. AB - The development of an inhibitor to transfused factor VIII (FVIII) is a serious treatment-related problem in haemophiliac children. The management of patients with high titre FVIII inhibitors is difficult, and immune tolerance induction (ITI) is the only method available for the eradication of these inhibitors. The results of the ITI regimen used at the Children's Hospital of Michigan Haemophilia Treatment Center are described and discussed. ITI was attempted in 14 children with severe haemophilia A (13 high responders, one low responder), with daily doses of FVIII alone. FVIII dosage was chosen according to the patient's historical peak inhibitor titre. ITI included three phases; induction phase, dose reduction phase and maintenance phase. During the first phase, the starting dose was 50 or 100 U kg-1 d-1; during the second phase the FVIII dosage was reduced gradually to 25 U kg-1 every other day according to the inhibitor titre, FVIII recovery and/or half-life study. In the third (maintenance) phase, the children received either prophylactic therapy or episodic therapy for 12 months. The inhibitor elimination was defined as the time taken to achieve a negative inhibitor assay with no anamnestic response and normal FVIII recovery and/or normal half-life. Immune tolerance was achieved in 11 of 14 patients (79%) patients within a median time of 6 months; two children are still on therapy, three failed ITI. We observed either failure or prolongation of immune tolerance if the historical peak titre or the maximum titre during ITI was >200 BU. The success rate of our low dose ITI regimen is not different from that reported by other investigators and the inhibitor elimination time is similar to some of the studies reported previously. PMID- 10792473 TI - Management of factor XI inhibitor for cardiac intervention: successful treatment with immunosuppressive therapy and plasma exchange. AB - Two cases with congenital homozygous factor XI deficiency developed a factor XI inhibitor following repeated plasma transfusions. Case 1 was given cyclophosphamide, intravenous immunoglobulin, and steroids. The factor XI inhibitor disappeared on day 103 and cardiac catheterization was performed without complications after giving fresh frozen plasma. Case 2 was effectively managed by plasma exchange for cardiac catheterization and surgery. However, after five plasma exchange procedures, the same plasma volume exchange was not effective in shortening the activated partial thromboplastin time (APTT). A significant heparin rebound occurred 4 h after heparin neutralization with protamine sulphate for which the patient needed to have a blood clot evacuated from around the heart. PMID- 10792475 TI - The orthopaedic status of severe haemophiliacs in Spain. AB - This paper provides an outline of the results obtained in a cross-sectional study conducted primarily with the aim of ascertaining orthopaedic complications in a group of young severe A and B haemophiliacs, the effects which these complications have on the medical resources used with these patients, and the impact of severe haemophilia on their quality of life. Its secondary aim was to link their current orthopaedic status to the type of treatment they had received prior to the study. Eleven Spanish hospitals took part in this study, monitoring 70 severe haemophiliacs (FVIII:C <2%) without inhibitors who had a mean age of 21.6 years and a median age of 22. Retrospective data collected from birth to the conclusion of the study were used and, for certain variables, data from the last 12 months. The type of treatment given had been on-demand treatment, together with prophylaxis of variable time periods, which in 32 cases (45.7%) were prolonged (>6 months). In 40 cases (57.1%) the patients underwent one or more periods of prophylaxis. Thirty-three patients (47.8%) had over 1000 days of administration of factors VIII and IX. The analysis of the total study group reveal an average of 348 bleeding episodes per patient. The findings of this study revealed that 84.3% of these patients suffer from articular complaints on the Gilbert scale, and 85.7% on the Pettersson scale. In addition, pain was reported in 16.1% of the joints, the most frequently affected being the ankle joints. Twenty-six patients (37%) had undergone orthopaedic surgery from the time of birth to the conclusion of the study. The quality of life of the severe haemophiliacs reviewed seems to have been affected. During the last 12 months, there were 216 outpatient haematological visits and 176 orthopaedic rehabilitation visits, as well as 12 radiological explorations and two hospitalizations. During these 12 months, medical expenditure totalled $55 473 per patient per year, the most important item being factor concentrates VIII/IX ($54 119 per patient per year). The type of treatment given to these patients (on demand and/or as secondary prophylaxis) was found to be incapable of preventing haemophilic arthropathy or guaranteeing an acceptable quality of life, although both the administration of coagulant factor concentrate to such patients and the financial resources dedicated to their treatment was very high. Consequently, a strategic change is to be considered for the prevention of haemophilic arthropathy, based on the introduction of replacement treatment involving continuous administration of factors VIII and IX in primary prophylaxis regimens from the first years of life. PMID- 10792474 TI - MRI findings in haemophilic joints treated with radiosynoviorthesis with development of an MRI scale of joint damage. AB - We hypothesized that magnetic resonance imaging (MRI) scans taken prior to radiosynoviorthesis may be predictive of response to the procedure in persons with haemophilia. Specifically, response would be inversely related to the severity of synovial hyperplasia. Radiosynoviorthesis was administered to 21 joints with recurrent haemorrhage (target joints). A detailed self-report of haemorrhage history, joint evaluation with scoring according to the World Federation of Haemophilia orthopaedic joint and pain scales, plain radiographs, and MRI studies of the joints were performed pre- and post-radiosynoviorthesis. To augment comparison of the MRI findings to those assessed using the Arnold Hilgartner and Pettersson scales, a provisional MRI scale for evaluation of haemophilic arthropathy was designed. We found the MRI findings prior to the procedure were not predictive of clinical response; independent of the severity of synovial hyperplasia, most joints bled less and showed improvement by the WFH orthopaedic score. There was generally no change in the severity of synovial hyperplasia after the procedure. We conclude that MRI evaluation is not routinely indicated prior to radiosynoviorthesis. PMID- 10792476 TI - Use of the dual force system to correct chronic knee deformities due to severe haemophilia. AB - In this study, the use of the dual force system to correct recent or relatively longstanding knee deformities in ten patients is described. (Nine of the patients had severe haemophilia and one had severe von Willebrand's disease.) The mean duration of deformity in these patients was 10 months. The mean range of movement at the affected knee joints increased from 50 degrees at pre-intervention to 110 degrees following 6 weeks of application of the dual force system. In nine of ten patients (90%) the residual flexion deformity ranged from 0 degrees to 10 degrees. The dual force system offers an easily affordable and effective means of correcting a flexion deformity of the knee joint in severely affected haemophilia and allied disorders. More extensive use of this technique in different centres is required to determine its place in the day-to-day management of such patients. PMID- 10792477 TI - Maintenance of safer sexual behaviours: evaluation of a theory-based intervention for HIV seropositive men with haemophilia and their female partners. AB - A theory-based HIV risk-reduction intervention was developed for HIV-positive men with haemophilia and their HIV-negative female romantic partners. The intervention was based on Prochaska and DiClemente's Transtheoretical Model which describes behaviour change as an incremental, stage-based process. The intervention targeted both communication about safer sex and safer sex behaviours (consistent condom use or abstinence from vaginal intercourse). A total of 255 males and 158 females from six funded haemophilia treatment centres or patient organizations (and 27 associated subsites) participated in the study. Baseline and follow-up (15 months after baseline) measures were administered to assess safer sexual behaviours, communication about safer sex and condom self-efficacy. A quasi-experimental, repeated measures design was utilized to compare two naturally occurring groups; those who received the full intervention package and those who received incomplete or no intervention components. Significant intervention effects for safer sex behaviours, communication about safer sex and condom self-efficacy were identified for the male participants, with those receiving the full intervention package demonstrating better outcomes at follow up. Women who received the full intervention package were more likely to report the use of a condom by their male partner during the last act of vaginal intercourse. PMID- 10792478 TI - Drug-induced and traumatic nail problems in the haemophilias. AB - Many persons with haemophilia suffer from HIV and receive highly active antiretroviral therapy. Three patients received indinavir and required surgery due to ingrown toenails. Two patients suffered from a traumatic subungual haematoma. The treatment protocol is described whereby the pressure exerted onto the germinal layer and the nail bed is relieved in order to alleviate pain and nail matrix damage. PMID- 10792479 TI - Diagnosis of two related carriers of severe haemophilia B with no family history. AB - Haemophilia B is an X-linked disease affecting 1 in 30 000 males. Carrier diagnosis is usually carried out only in female relatives of haemophilic males, and the likelihood of discovering a carrier without a haemophilic male is very low. In this report we present the cases of two related women without a family history of haemophilia who were diagnosed as haemophilia B carriers. Following a minor haemorrhage in the proband, she and her mother were thought to be haemophilia B carriers because of a low factor IX level (16 and 23 IU dL-1, respectively; normal values >50 IU dL-1). The non-sense mutation C31118T, which is associated with severe haemophilia B, was detected in both women. This allowed us to diagnose them as being definite carriers of severe haemophilia B and give appropriate genetic counselling. PMID- 10792480 TI - Morphological and immunophenotypic variations in malignant melanoma. AB - A variety of cytomorphological features, architectural patterns and stromal changes may be observed in malignant melanomas. Hence, melanomas may mimic carcinomas, sarcomas, benign stromal tumours, lymphomas, plasmacytomas and germ cell tumours. Melanomas may be composed of large pleomorphic cells, small cells, spindle cells and may contain clear, signet-ring, pseudolipoblastic, rhabdoid, plasmacytoid or balloon cells. Various inclusions and phagocytosed material may be present in their cytoplasm. Nuclei may show bi- or multi-nucleation, lobation, inclusions, grooving and angulation. Architectural variations include fasciculation, whorling, nesting, trabeculation, pseudoglandular/pseudopapillary/pseudofollicular, pseudorosetting and angiocentric patterns. Myxoid or desmoplastic changes and very rarely pseudoangiosarcomatous change, granulomatous inflammation or osteoclastic giant cell response may be seen in the stroma. The stromal blood vessels may exhibit a haemangiopericytomatous pattern, proliferation of glomeruloid blood vessels and perivascular hyalinization. Occasionally, differentiation to nonmelanocytic structures (Schwannian, fibro-/myofibroblastic, osteocartilaginous, smooth muscle, rhabdomyoblastic, ganglionic and ganglioneuroblastic) may be observed. Typically melanomas are S100 protein, NKIC3, HMB-45, Melan-A and tyrosinase positive but some melanomas may exhibit an aberrant immunophenotype and may express cytokeratins, desmin, smooth muscle actin, KP1 (CD68), CEA, EMA and VS38. Very rarely, neurofilament protein and GFAP positivity may be seen. PMID- 10792481 TI - Cytokeratin profiles of the thymus and thymomas: histogenetic correlations and proposal for a histological classification of thymomas. AB - AIMS: Since cytokeratins (CKs) are useful as differentiation markers for histogenetic and classification studies, we investigated the CK profiles of the thymus and thymomas in an attempt to understand the histogenetic correlation and to propose a histological classification. METHODS AND RESULTS: Nine thymuses and 34 thymomas were immunostained for various CKs of different molecular weights and involucrin. Based on cytomorphology and histoarchitecture, thymomas were classified into spindle cell (SC), small polygonal cell (SPC), mixed, organoid, large polygonal cell (LPC) and squamoid (SQ) thymomas for compiling CK profiles. The thymus was shown to comprise four epithelial compartments, each expressing a different CK profile. Different histological types of thymoma expressed different CK profiles. By correlating the CK profiles of the thymus and thymoma, SPC, SC and LPC thymomas appeared to be related to subcapsular, medullary and cortical cells, respectively. Organoid thymoma recapitulated the structure and CK profile of the normal thymus, while SQ thymoma acquired additional squamous type CK. The applicability and usefulness of the proposed histological classification were evaluated on 147 thymomas by correlating the results with their invasive behaviour. One hundred and thirty-nine cases (95%) could be classified and different histological types correlated strongly with their invasive behaviour. CONCLUSIONS: The thymus is a complex epithelial organ composed of heterogeneous cell types giving rise to various related histological types of thymoma. The results of the CK profile study supports the proposed histological classification, which is pathologically applicable and clinically useful in correlating with invasiveness. This cytomorphological classification, supported by the CK expression patterns, is comparable to Muller-Hermelink classification and the new WHO histological classification except that a separate group of SPC thymoma expressing only CK14 and CK19 was identified and separated from mixed thymoma. PMID- 10792482 TI - Expression of thyroid transcription factor-1 in pulmonary and extrapulmonary small cell carcinomas and other neuroendocrine carcinomas of various primary sites. AB - AIMS: The thyroid transcription factor-1 (TTF-1) is a highly specific immunohistochemical marker for the identification of pulmonary adenocarcinomas and non-neuroendocrine large cell carcinomas, especially in patients presenting with metastatic carcinomas of unknown primary site. In this study we tested if anti-TTF-1 can also be used to verify a pulmonary origin of neuroendocrine carcinomas, placing emphasis on the discrimination of pulmonary small cell carcinomas (SCCs) from extrapulmonary SCCs and the distinction of SCCs from Merkel cell carcinomas of the skin. METHODS AND RESULTS: We studied 37 pulmonary SCCs, 15 SCCs of extrapulmonary origin, 4 pulmonary large cell neuroendocrine carcinomas (LCNECs), four extrapulmonary LCNECs, six medullary thyroid carcinomas, 16 Merkel cell carcinomas, and a total of 32 carcinoids/low-grade neuroendocrine carcinomas of pulmonary (12 cases) and extrapulmonary (20 cases) origin. Using the commercially available monoclonal antibody 8G7G3/1, TTF-1 was immunohistochemically detectable in 81% of pulmonary SCCs but also in 80% of extrapulmonary SCCs. Furthermore, anti-TTF-1 showed a positive staining in 50% of all pulmonary carcinoids, in one gastric carcinoid, in 2/4 of pulmonary, and 1/4 of extrapulmonary LCNECs. All medullary thyroid carcinomas were also TTF-1 positive. Merkel cell carcinomas were consistently TTF-1-negative. CONCLUSIONS: Our results suggest that in contrast to non-neuroendocrine carcinomas, anti-TTF-1 cannot be used to prove or to exclude a pulmonary origin of SCCs or LCNECs of unknown or uncertain primary site. Therefore, before using anti-TTF-1 as a marker for pulmonary carcinomas one should be sure to have excluded SCC and LCNEC. On the other hand, anti-TTF-1 might be used to specifically discriminate SCCs of various origin from Merkel cell carcinomas. PMID- 10792484 TI - Transitional cell carcinomas of the ovary and bladder are immunophenotypically different. AB - AIMS: Transitional cell carcinoma (TCC) of the ovary is a subtype of ovarian cancer whose main characteristic is its histological resemblance to TCC of the bladder. Thrombomodulin (TM), a surface glycoprotein commonly expressed in normal and neoplastic urothelium, has been proven to be a good marker for TCC of the bladder. To better define the phenotype of TCC of the ovary, we investigated TM, cytokeratin 20 and carcinoembryonic antigen (CEA) expression in 15 TCCs of the ovary and compared their phenotype with that of 20 TCCs of the bladder, and 20 serous and 10 endometrioid carcinomas of the ovary. METHODS AND RESULTS: Immunostaining was performed on formalin-fixed, paraffin-embedded tissue sections using the avidin-biotin-peroxidase complex method. All 20 TCCs of the bladder stained for TM and cytokeratin 20, and 13 stained for CEA. None of the TCCs of the ovary reacted for TM or cytokeratin 20, and only two expressed CEA. All of the serous and endometrioid carcinomas were negative for TM and cytokeratin 20. CEA positivity was observed in two of the serous carcinomas, but in none of the endometrioid carcinomas. CONCLUSION: The immunophenotype of TCC of the ovary is similar to that of other surface carcinomas of the ovary, but differs from that of TCC of the bladder. Since immunohistochemical procedures are often used in the diagnosis and classification of both primary and metastatic tumours, it is important to be aware of these differences in immunophenotype. PMID- 10792483 TI - Prognostic and immunohistochemical validation of the capella classification of pancreatic neuroendocrine tumours: an analysis of 82 sporadic cases. AB - AIMS: To study the clinical outcome of 82 cases of pancreatic neuroendocrine tumours classified according to the recent histological and prognostic classification of Capella. METHODS AND RESULTS: Eighty-two surgical cases of pancreatic neuroendocrine tumours were examined histologically with immunohistochemical staining of paraffin sections using streptavidin-biotin complex and application of antibodies against chromogranin A and 10 hormonal peptides. Classification in four groups correlated with long follow-up and outcome of these cases. Histological examination showed 30 group I, four group II, 41 group III and seven group IV tumours. Twenty-one (70%) of group I tumours were insulinomas, whereas 25% of group III tumours were glucagonomas and 25% were unclassified. Most group IV tumours were unclassified, showing no immunohistochemical staining with any of the 10 hormonal peptides tested. Outcome was clearly correlated with tumour group. Among the 14 patients who died of the disease, four had group IV and 10 group III tumours. Thus, unclassified asymptomatic tumours without immunohistochemical staining had a poorer prognosis than asymptomatic tumours with staining. CONCLUSION: This study validates the Capella classification as easy to apply and useful in predicting clinical outcome. PMID- 10792485 TI - Acinar morphology in colorectal cancer is associated with survival but is not an independent prognostic variable. AB - AIMS: Microacinar morphology has been reported as a stage-independent prognostic variable for colorectal cancer. We have undertaken a retrospective analysis to test this observation in patients with colonic as well as rectal adenocarcinoma, and to assess interobserver agreement for identifying microacinar morphology. METHODS AND RESULTS: One hundred and eighty patients with colorectal cancer, 144 colonic and 36 rectal, were assigned to either macroacinar (144) or microacinar (36) groups. There was excellent correlation between two observers examining the colorectal cancers independently (kappa statistic, kappa = 0.87). Patients with microacinar tumours had a significantly reduced median survival compared with macroacinar lesions (46 and 87 months, respectively, P = 0.022) and this morphology was significantly associated with higher Dukes' stage (P = 0.007). Microacinar morphology lacked statistical significance when examined in a multivariate analysis with other prognostic variables, both when colonic and rectal tumours were examined either separately or as a combined colorectal group, but small numbers of rectal tumours in this study make it difficult to draw useful conclusions about microacinar morphology in rectal adenocarcinoma. CONCLUSIONS: Microacinar morphology was strongly associated with poor prognosis when examined in isolation but was not found to be an independent factor when examined with other prognostic variables. PMID- 10792486 TI - Malignant angiomyolipoma of the liver: a hitherto unreported variant. AB - AIMS: After their original recognition in the kidney, angiomyolipomas (AMLs) have been reported in the liver for more than 20 years. In the kidney, five cases of malignant AML have been reported. We report the first case of malignant hepatic AML. METHODS AND RESULTS: A 70-year-old female patient presented with abdominal discomfort. Clinical examination revealed a palpable liver. CT scan showed a polymorphous hypervascular lesion in the right liver lobe. A biopsy was taken and resulted initially in a differential diagnosis between a hepatocellular carcinoma, a metastatic tumour (possibly of renal origin) and angiomyolipoma (AML). After immunohistochemistry, a hepatic AML was suggested, given the immunoreactivity for HMB45/NKIC-3. The mass was resected 5 years later because of relapsing abscess formation. Gross examination of the resection specimen showed a focally encapsulated brown mass with focal necrosis. Microscopic examination showed a tumour growing in sheets, separated by sinusoidal-like vessels. Most of the tumour cells had a large, polygonal, clear cytoplasm, often with eosinophilic condensation around the nucleus. There was prominent vascular invasion. Immunohistochemistry (reactivity for HMB-45, NKIC-3, S100 and alpha smooth muscle actin, negativity for cytokeratin and vimentin) and electron microscopy confirmed the diagnosis of monomorphic epithelioid AML with prominent vascular invasion. Seven months after tumour resection, the patient died of recurrent disease. CONCLUSIONS: This case highlights the importance of immunohistochemistry and electron microscopy in diagnosing this type of tumour. Possibly, in the past, malignant AML of the liver has been misdiagnosed as HCC. PMID- 10792487 TI - Monotypic epithelioid angiomyolipoma of the liver. AB - AIMS: Monotypic epithelioid angiomyolipoma is a recently recognized renal tumour, which is composed purely of epithelioid cells coexpressing markers of both smooth muscle differentiation and melanogenesis (HMB45). We report here the first case of monotypic epithelioid angiomyolipoma arising in the liver. CASE DETAILS: A 30 year-old woman without tuberous sclerosis complex (TSC) was incidentally found to have a hepatic mass by ultrasonography. Grossly, the resected tumour showed a nodule-in-nodule appearance, with large areas of haemorrhagic necrosis. Microscopically, the tumour was composed of pleomorphic epithelioid cells with clear, eosinophilic cytoplasm. Neither adipocytes nor abnormal vessels were recognized in the tumour. Immunohistochemically, the tumour cells were strongly positive for HMB45 and S100 protein, focally positive for desmin, vimentin and smooth muscle actin, and negative for epithelial markers (cytokeratins, EMA). Ultrastructural analysis showed numerous dense granules with some striated ones resembling melanosomes, myofilaments and pinocytic vesicles in the cytoplasm. Molecular analysis showed no allelic loss of the TSC2 region or 12 other chromosomal regions. The patient is free of disease over 1 year after the operation. CONCLUSION: We consider that this hepatic tumour is closely related to angiomyolipoma, and a counterpart of renal monotypic epithelioid angiomyolipoma. PMID- 10792489 TI - Towards a histogenetic classification of thymic epithelial tumours? PMID- 10792488 TI - Renal lesions in von Hippel-Lindau disease: immunohistochemical expression of nephron differentiation molecules, adhesion molecules and apoptosis proteins. AB - AIMS: Renal lesions in von Hippel-Lindau disease comprise clear cell simple cysts, atypical cysts and carcinomas. Although histological and molecular studies suggest that cystic lesions may represent precursors of carcinomas, there is no detailed phenotypic evidence of their relationship. METHODS AND RESULTS: To investigate such a possible relationship between cystic lesions and solid carcinomas, we studied the pathological and immunohistochemical features of 328 lesions of 33 kidneys originating from 23 patients with von Hippel-Lindau disease, using a panel of antibodies directed against cytoskeleton proteins, cell surface proteins, integrin subunits, adhesion molecules, lectins, and apoptosis and proliferation markers. Solid carcinomas (n = 175) were all of clear cell type and mostly nuclear grade 1. Cystic lesions (n = 138) consisted of cystic clear cell carcinomas (n = 15), atypical cysts (n = 20) and simple cysts (n = 103). Clear cells of the simple cysts, atypical cysts and solid carcinomas coexpressed cytokeratins (CK8, CK19) and vimentin, and expressed a similar pattern of tubular markers (CD24, tetraglonolobus), integrin subunits (alpha3, alpha5, alpha6, alphav, beta1) and cell adhesion molecules (ICAM 1, VCAM 1). In all lesions studied, proliferation rate (MIB1 index) was low, and apoptosis marker expression (fragmented DNA, p53, bcl-2) inconspicuous. CONCLUSIONS: Phenotypic alterations found in solid renal cell carcinomas are already present in simple and atypical renal cysts of von Hippel-Lindau disease. PMID- 10792490 TI - Collagenous spherulosis versus shadow cell differentiation in endometrioid adenocarcinoma PMID- 10792491 TI - Clear cell change in metastatic colonic adenocarcinoma. PMID- 10792492 TI - Hybrid neurofibroma/schwannoma versus schwannoma with Antoni B areas. PMID- 10792493 TI - Pleomorphic lipoma with pseudopapillary structures: a pleomorphic counterpart of pseudoangiomatous spindle cell lipoma. PMID- 10792495 TI - Forced expression of terminal deoxynucleotidyl transferase in fetal thymus resulted in a decrease in gammadelta T cells and random dissemination of Vgamma3Vdelta1 T cells in skin of newborn but not adult mice. AB - The repertoire of lymphocyte receptor genes encoded in a germline is further diversified by a number of processes, including the template-independent addition of nucleotides (N regions) by means of terminal deoxynucleotidyl transferase (TdT). Normally, mouse gammadelta T cells in the early fetal thymus, whose T-cell receptor (TCR) genes lack N regions and are encoded by Vgamma3-Jgamma1 and Vdelta1-Ddelta2-Jdelta2 with canonical junctions (invariant Vgamma3Vdelta1), are thought to be the precursors of dendritic epidermal T cells (DETC). We generated mutant mice whose endogenous TdT promoter was replaced with the lck promoter through homologous recombination. These mutant mice expressed TdT in fetal thymus, had abundant N regions and infrequent canonical junctions in gamma and delta rearrangements, and showed a decreased number of gammadelta T cells. Various Vgamma3Vdelta1 T cells, most of which had N regions in their TCR genes, were found to disseminate in the skin of newborn mutant mice, whereas normal numbers of DETCs with the invariant Vgamma3Vdelta1 rearrangement were observed in adult mutants. These data demonstrate that the regulation of TdT expression during fetal development is important for the generation of gammadelta T cells, and that Vgamma3Vdelta1 T cells, which have various junctional sequences in their TCR genes, randomly disseminate in skin, but invariant Vgamma3Vdelta1 T cells have a great advantage for proliferation in skin. PMID- 10792494 TI - Interaction of merosin (laminin 2) with very late activation antigen-6 is necessary for the survival of CD4+ CD8+ immature thymocytes. AB - The laminin alpha2-chain is a component of merosin, a member of the laminin family molecules, which is mainly expressed in the basement membranes of striated muscle. It is known that laminin alpha2 gene (lama2) null mutant mice (dy3k/dy3k) exhibit congenital muscular dystrophy (CMD). Because the laminin alpha2-chain is also expressed in the thymus, the role of merosin in the thymus was examined. In association with the onset of muscular dystrophy, CD4+ CD8+ double-positive (DP) thymocytes disappear by apoptotic cell death, while CD4+ CD8- or CD4- CD8+ thymocytes remain. In order to study the mechanisms leading to the selective death of DP cells in the absence of merosin, the role of the interaction between very late activation antigen-6 (VLA-6), a candidate merosin ligand in the thymus, and merosin was examined. The in vitro survival of thymocytes from normal mice was maintained by the addition of either anti-VLA-6 monoclonal antibodies (mAbs) or merosin. Furthermore, when the normal thymocytes were cultured on thymic epithelial cell lines, viable DP cell recoveries on wild-type epithelial cells were better than on cells from null mutant mice. The results suggest that DP cells are more sensitive to an uncharacterized apoptotic death signal, and that survival is supported by the interaction between VLA-6 and merosin. PMID- 10792496 TI - Decreased expression of FcgammaRIII (CD16) by gammadelta T cells in patients with rheumatoid arthritis. AB - Some gammadelta T cells express a receptor for the Fc portion of immunoglobulin G (FcgammaRIII - CD16). The relevance of this Fc receptor to gammadelta T-cell function is at present unclear. Our previous studies have shown that gammadelta T cells express activation markers in patients with rheumatoid arthritis (RA). In this study we have examined the relative proportions of CD16+ gammadelta T cells in the blood and synovial fluid of these patients compared with control blood. CD16+ gammadelta T cells from RA patients were significantly reduced in synovial fluid compared with the circulation. That this was due to blocking of antibody binding to CD16 was unlikely as treatment of blood gammadelta T cells with RA synovial fluid (known to contain immune complexes) failed to alter expression of CD16. Treatment of blood gammadelta T cells with phytohaemagglutinin in vitro, resulted in a time-dependent decrease in expression of CD16, with a concomitant increase in expression of human leucocyte antigen-DR, at the single cell level. We conclude that expression of CD16 by gammadelta T cells is lost in the synovial compartment as the result of activation. PMID- 10792497 TI - Characterization of the porcine gammadelta T-cell receptor structure and cellular distribution by monoclonal antibody PPT27. AB - The T-cell receptor (TCR) is the critical structure involved in antigen recognition of T lymphocytes. Although the pig has a large proportion of circulating T lymphocytes bearing the gammadelta TCR, their study has been impeded due to the lack of specific antibodies. Here a monoclonal antibody (mAb) PPT27 directed to gammadelta TCR is described. Flow cytometry analyses showed that the mAb recognized a subset of T lymphocytes of which the majority expressed no CD2, CD4 and CD8 whilst the minority bore CD2 and CD8. The mAb precipitated a protein of 86 000 MW under non-reducing conditions and a doublet of 43 000 MW under reducing conditions from peripheral blood T lymphocytes lysed in nonidet P 40 buffer, whilst it precipitated the CD3-TCR complex from the cells lysed in digitonin. Further analysis revealed that the antibody recognized the majority, but not all, of the gammadelta T cells, suggesting that there may be more isotypes of gammadelta TCR than currently believed. The antibody was unable to stimulate gammadelta T cells to proliferate in vitro, suggesting that these cells are activated by a different activation mechanism from that of alphabeta T cells. PMID- 10792498 TI - Native IgG2a(b) is barely antigenic to major histocompatibility complex class II restricted T cells owing to inefficient internalization by professional antigen presenting cells. AB - Peptide epitopes derived from immunoglobulin variable regions represent tumour specific antigens on B-cell neoplasms and can be recognized by syngeneic, major histocompatibility complex (MHC) class II-restricted T cells. Immunoglobulin peptide/MHC class II complexes may also be involved in autoimmunity and CD4+ T cell-mediated B-cell regulation. Thus, the IgG2a(b) H-chain allopeptide gamma2a(b) 435-451 presented on I-Ad mimics the epitope implicated in herpes simplex virus-induced autoimmune stromal keratitis and is the target of T helper 1 (Th1) clones that suppress IgG2a(b) production in vivo. We here report that spleen and thymus cells constitutively present the autologous gamma2a(b) epitope to a gamma2a(b) 435-451/I-A(d) reactive T-cell hybridoma as a function of the animal housing conditions (specific pathogen-free or not) and the serum levels of IgG2a(b). Constitutive presentation in the spleen was predominantly performed by dendritic cells. Whereas spleen cells poorly presented native IgG2a(b) to a gamma2a(b) 435-451/I-A(d) reactive T-cell hybridoma, IgG2a(b) in the form of immune complexes were presented > 200-fold more efficiently owing to internalization via low-affinity FcgammaR on macrophages. The antigenicity could also be improved by homotypic aggregation and by targeting IgG2a(b) to complement receptors on the A20 B-cell lymphoma. Mice without detectable IgG2a(b)-containing immune complexes typically exhibited minimal constitutive presentation. Nevertheless, native IgG2a(b) can sensitize antigen-presenting cells in vivo, as mice that were devoid of immune complexes and carried an IgG2a(b)-producing tumour did present constitutively, even at physiological IgG2a(b) serum levels. Whereas the amounts of IgG released from most B-cell lymphomas may be too low to allow spontaneous priming of tumour-specific MHC class II-restricted T cells, administration of tumour immunoglobulin in aggregated form might improve the efficacy of idiotype vaccination. PMID- 10792499 TI - Interleukin-10 does not affect phagocytosis of particulate antigen by bone marrow derived dendritic cells but does impair antigen presentation. AB - Dendritic cells (DC) are important initiators of an immune response so understanding the factors controlling antigen acquisition and presentation has important consequences for the use of these cells in vaccines and other forms of immunotherapy. We investigated the factors that influence phagocytosis by immature bone marrow-derived DC (BMDC) and the effect of interleukin-10 (IL-10) on this process. Two sizes of fluorescent particles and recombinant bacillus Calmette-Guerin expressing the green fluorescent protein (rBCG) were used as particulate antigens. The percentage of cells taking up the antigen was found to be dependent on the size and dose of the particles, and the length of exposure to them. BMDC exposed to IL-10 at various concentrations for different periods exhibited no distinguishable change in antigen uptake. However, if BMDC treated with IL-10 and rBCG were then exposed to a second dose of particulate antigen, uptake was increased compared with those BMDC not treated with IL-10. The expression of major histocompatibility complex class II, CD80, CD86 and CD11c by BMDC after phagocytosing rBCG or inert beads, was inhibited when the BMDC were pretreated with IL-10. In contrast, the expression of CD25 was increased. BMDC that had taken up BCG or purified protein derivative (PPD) were able to stimulate primed T-cell proliferation but this was severely inhibited if the BMDC were cultured with IL-10 before exposure to the antigen. This work suggests that although IL-10 does not affect the phagocytic capacity of BMDC, it does inhibit maturation of the cells and consequently, T-cell activation. PMID- 10792500 TI - Fibronectin synthesis by activated T lymphocytes: up-regulation of a surface associated isoform with signalling function. AB - Fibronectin (FN) is a major constituent of the extracellular matrix. We now provide evidence for a surface-associated isoform of FN that is synthesized by T cells upon activation. The T-cell-derived FN has an unusual splice pattern: an additional domain, EDB, is produced whereas sequences within another domain, IIICS, are spliced out. CS1, the binding domain for very late antigen-4 (VLA-4), however, is still generated. To study the potential function of surface associated FN its synthesis was down-regulated by an antisense oligonucleotide, then proliferation of T cells was induced by cross-linked anti-CD3. Proliferation was reduced as was expression of CD25. Moreover, when T cells were cultured in high density, the synthetic peptide QILDVPST, corresponding to CS1, inhibited proliferation, as did antibodies to VLA-4. We propose that surface-associated FN is a ligand for VLA-4, which by binding to VLA-4 on an adjacent cell, provides a costimulatory signal, thus sustaining T-cell proliferation. PMID- 10792501 TI - Interferon-gamma up-regulates intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 and recruits lymphocytes into the vagina of immune mice challenged with herpes simplex virus-2. AB - Lymphocyte recruitment into tissues involves interactions between adhesion molecules on vascular endothelial cells and corresponding ligands on the lymphocyte surface. In the present study we investigated the expression of four endothelial addressins in the vagina and their possible up-regulation by interferon-gamma (IFN-gamma) in immune mice after vaginal challenge with herpes simplex virus type 2. The adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were minimally expressed in the vagina of non-immune mice with or without vaginal challenge and in immune mice before challenge, but both were up-regulated by IFN-gamma, directly or indirectly, in immune mice after challenge. Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was detected in most vaginas but was not up-regulated by IFN-gamma in immune mice after virus challenge. E-selectin was not detected in any vaginas. The results suggest that ICAM-1 and VCAM-1 may be involved in rapid, IFN-gamma-mediated recruitment of lymphocytes to the vaginal mucosal of immune mice after local virus challenge. PMID- 10792502 TI - Antibody cross-linking of human CD9 and the high-affinity immunoglobulin E receptor stimulates secretion from transfected rat basophilic leukaemia cells. AB - Previous studies have shown that antibody cross-linking of the tetraspanin protein CD9 stimulates the degranulation of platelets and eosinophils, although the mechanism of activation is unclear. In this work we transfected human CD9 into the rat basophilic leukaemia (RBL-2H3) cell line and studied the stimulation of secretion from these cells in response to a panel of anti-CD9 antibodies. Intact immunoglobulin G1 (IgG1) antibodies activated transfected cells whereas F(ab')2 fragments of antibody and an intact IgG2a did not. Stimulation of secretion was inhibited by co-incubation with monomer murine immunoglobulin E (IgE) but not with an IgG1 isotype control, indicating that the response involves the endogenous high-affinity IgE receptor (FcepsilonRI). The anti-CD9 antibody activation curve was biphasic, and supraoptimal antibody concentrations stimulated little or no degranulation, indicating that multivalent binding of human CD9 molecules is necessary for the formation of an active complex with rat FcepsilonRI. Immunoprecipitation of FcepsilonRI under mild detergent conditions co-precipitated CD9, suggesting the presence of pre-existing complexes of CD9 and FcepsilonRI that could be activated by antibody cross-linking. These data are further evidence that tetraspanins are involved in FcepsilonRI signalling and may reflect the participation of tetraspanins in the formation of complexes with other membrane proteins that use components of Fc receptors for signal transduction. PMID- 10792503 TI - Nuclear factor-kappaB and caspases co-operatively regulate the activation and apoptosis of human macrophages. AB - Accumulating evidence suggests that macrophages function as major effector cells in the pathological process of various human diseases. We examined here the role of nuclear factor-kappaB (NF-kappaB) and caspases in the regulation of activation and apoptosis of macrophages. Activation of the human monoblastic leukaemia cell line, U937, by phorbol 12-myristate 13-acetate (PMA) increased the expression of CD14/CD86, and cytokine production. PMA stimulation also increased the expression of both pro-caspase-8 and pro-caspase-3 in U937, but not apoptosis or intracellular caspase-3 activity. PMA also increased the expression of X chromosome-linked inhibitor of apoptosis protein (XIAP) in U937, suggesting an inhibitory action for XIAP on the caspase cascade in PMA-stimulated U937. Electrophoretic mobility shift assay (EMSA) showed a significant increase of nuclear NF-kappaB activity in PMA-stimulated U937. When a potent NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), was added to U937 cell culture in the presence of PMA, apoptosis was triggered by activation of caspase-3, which was induced by caspase-8 activation. XIAP expression was markedly suppressed in PMA-treated U937 in the presence of PDTC. The inhibitors of caspase-8 and caspase 3 mostly inhibited apoptosis of U937 treated with PMA in the presence of PDTC. Furthermore, a phenotype of U937 treated with PMA and PDTC in the presence of caspase inhibitor was almost identical to that of unstimulated U937. Our results suggest that the signalling pathways involved in the activation and apoptosis of human macrophages could be co-operatively regulated by the use of NF-kappaB and caspase inhibitors, thus enabling the control of macrophage function and number. PMID- 10792504 TI - Transcriptional and post-transcriptional regulation of monocyte chemoattractant protein-3 gene expression in human endothelial cells by phorbol ester and cAMP signalling. AB - Monocyte chemoattractant protein-3 (MCP-3) is one of the most broadly active chemokines, potentially inducing chemotaxis of all leucocytic cells. In the present study, we examined the regulation of MCP-3 mRNA and protein production in endothelial cells by protein kinase C (PKC) activator, phorbol 12-myristate 13 acetate (PMA) and cAMP signalling. On stimulation of endothelial cells with 10 nM PMA, MCP-3 mRNA increased to 300-fold the basal level at 3 hr and rapidly declined to 0.2-fold the basal level at 24 hr. PMA-induced MCP-3 mRNA and protein production of human endothelial cells were partially inhibited by pretreatment with the adenylate cyclase activator, forskolin, or membrane-permeable cAMP derivative. The PMA-induced MCP-3 mRNA increase was almost abrogated when cells were pretreated with cycloheximide (CHX). Forskolin inhibited the transcription of PMA-induced MCP-3 gene expression. Following PMA stimulation for 3 hr, subsequent addition of actinomycin D suppressed the rapid decay of PMA-induced MCP-3 mRNA. These results suggest that PMA induces the transcriptional activation of the MCP-3 gene through de novo protein synthesis and the rapid decay of PMA induced MCP-3 mRNA through de novo synthesis of adenosine/uridine (AU)-rich element binding proteins and cAMP signalling inhibits the PMA-induced transcriptional activation of the MCP-3 gene expression. PMID- 10792505 TI - Up-regulation of beta-chemokines and down-modulation of CCR5 co-receptors inhibit simian immunodeficiency virus transmission in non-human primates. AB - A non-cognate mechanism of protection against human immunodeficiency virus-1 (HIV 1) infection involves up-regulation of beta-chemokines, which bind and may down modulate the CCR5 co-receptors, thereby preventing transmission of M-tropic HIV 1. The objective of this investigation was to evaluate this mechanism in vivo in non-human primates. Rhesus macaques were immunized by a modified targeted lymph nodes (TLN) route with recombinant simian immunodeficiency virus (SIV) glycoprotein 120 (gp120) and p27 in alum, and adsorbed recombinant granulocyte macrophage colony-stimulating factor (GM-CSF) with either interleukin (IL)-2 or IL-4. Immunization induced significant increases in the concentrations of CD8 cell-derived suppressor factor (CD8-SF), regulated on activation normal T cells expressed and secreted (RANTES), macrophage inflammatory protein (MIP)-1alpha and MIP-1beta, and down-modulation of the proportion of cells expressing CCR5 (r = 0.737, P<0.05). The macaques were then challenged with SIVmac 220 by the rectal mucosal route. The plasma SIVmac RNA showed a significant inverse correlation with the CD8-SF or the concentration of the three beta-chemokines (r = 0.831 and 0.824, P<0.01), but a positive correlation between the proportion of CCR5+ cells and SIVmac RNA (r = 0.613, P = 0.05). These results demonstrate for the first time in vivo that immunization up-regulates beta-chemokines, which may down modulate CCR5 co-receptors, and both functions are significantly correlated with the viral load. Hence, the non-cognate beta-chemokine-CCR5 mechanism should be considered as complementary to specific immunity in vaccination against HIV. PMID- 10792506 TI - Intercellular communication in the immune system: differential expression of connexin40 and 43, and perturbation of gap junction channel functions in peripheral blood and tonsil human lymphocyte subpopulations. AB - The distribution and function of connexins (integral membrane proteins assembled into gap junction intercellular communication channels) were studied in human lymphocyte subpopulations. The expression of mRNA encoding connexins in peripheral blood and tonsil-derived T, B and natural killer (NK) lymphocytes was examined. Connexin43 (Cx43) mRNA was expressed in peripheral blood and tonsil lymphocytes, but Cx40 mRNA expression was confined to tonsil-derived T and B lymphocytes; Cx26, Cx32, Cx37 and Cx45 were not detected by reverse transcription polymerase chain reaction (RT-PCR). Western blot analysis also demonstrated the presence of Cx40 and Cx43 proteins in T and B lymphocytes in a manner coincidental to the mRNA detection. Stimulation in vitro of T and B lymphocytes with phytohaemagglutinin (PHA) and lipopolysaccharide (LPS), respectively, increased Cx40 and Cx43 protein expression. Flow cytometric analysis, using antibodies to extracellular loop amino acid sequences of connexins, confirmed the surface expression of connexins in all lymphocyte subpopulations. Assembly of connexins into gap junctions providing direct intercellular channels linking attached lymphocytes was demonstrated by using a dye transfer technique. The exchange of dye between lymphocytes was inhibited by a connexin extracellular loop mimetic peptide and alpha-glycyrrhetinic acid, two reagents that restrict intercellular communication across gap junctions. Dye coupling occurred between homologous and heterologous co-cultures of T and B lymphocytes, and was not influenced by their stimulation with PHA and LPS. The connexin mimetic peptide caused a significant decrease in the in vitro synthesis of immunoglobulin M (IgM) by T- and B-lymphocyte co-cultured populations in the presence or absence of stimulation by PHA. The results identify connexins as important cell surface components that modulate immune processes. PMID- 10792507 TI - Replenishment of RANTES mRNA expression in activated eosinophils from atopic asthmatics. AB - Eosinophils have been shown to express the gene encoding regulated upon activation, normal T-cell expressed and secreted (RANTES), a potent eosinophilotactic chemokine. RANTES protein expression in eosinophils has previously been shown to be up-regulated by a number of agonists, including complement-dependent factors (C3b/iC3b) and interferon-gamma (IFN-gamma). We hypothesized that gene expression of RANTES is regulated in these cells by eosinophil-specific agonists. We analysed RANTES mRNA expression by reverse transcription polymerase chain reaction (RT-PCR) in human peripheral blood eosinophils obtained from mild atopic asthmatics following stimulation over time. In resting eosinophils, a low level of RANTES mRNA was found to be constitutively expressed in all the atopic donors tested in this study (n = 6). Following stimulation with C3b/iC3b (serum-coated surfaces), eosinophils released measurable levels of RANTES, while sustained transcript expression was detected for up to 24 hr of stimulation. In contrast, IFN-gamma (5 ng/ml) transiently and significantly (P<0.05, n = 3) depleted relative amounts of RANTES PCR product (compared with beta2-microglobulin) after 1-4 hr of stimulation. RANTES transcript was again detectable after 24 hr of IFN-gamma incubation, suggesting that the pool of RANTES mRNA had been replenished. Other eosinophil-active cytokines, interleukin-3 (IL-3), IL-4, IL-5 and granulocyte-macrophage colony stimulating factor, did not appear to modulate RANTES mRNA expression after 1 hr of incubation. The effect of IFN-gamma on RANTES mRNA was reversed by cycloheximide, suggesting that IFN-gamma may act by increasing the rate of translation of RANTES mRNA. These findings indicate that IFN-gamma may induce a rapid and transient effect on the translation and replenishment of RANTES mRNA in eosinophils. This novel observation supports the notion that eosinophils have the potential to replenish their stored and released bioactive proteins. PMID- 10792508 TI - Analysis of histamine-producing cells at the late phase of allergic inflammation in rats. AB - To identify histamine-producing cells at the late phase of allergic inflammation, the expression of L-histidine decarboxylase (HDC) was examined in the infiltrating leucocytes in the inflammatory locus. HDC activity and HDC mRNA levels in the infiltrating leucocytes in the pouch fluid of the immunized rats (that were injected with the antigen solution into the air pouch) were increased compared with those in the infiltrating leucocytes of the non-immunized rats. When infiltrating leucocytes collected 8 hr after antigen injection were cultured, histamine production by the cells from the immunized rats was higher than that from the non-immunized rats. In situ hybridization of HDC mRNA revealed that almost all the infiltrating leucocytes of the immunized rats, 4 hr after injection of the antigen, expressed HDC mRNA with high intensity, while those of the non-immunized rats showed only a weak intensity of HDC mRNA. In the immunized rats, approximately 90% of leucocytes infiltrating in the pouch fluid at 4 hr were neutrophils and 8% were monocytes/macrophages. Neither mast cells nor basophils were detected in the infiltrating leucocytes. When rat peritoneal neutrophils were incubated in the presence of 12-O-tetradecanoylphorbol 13 acetate, histamine production was significantly increased. These findings suggest that the leucocytes, mainly neutrophils, infiltrating at the inflammatory locus are responsible for histamine production at the late phase of allergic inflammation. PMID- 10792509 TI - Bacterial lipopolysaccharide acts as an adjuvant to induce autoimmune arthritis in mice. AB - We investigated the ability of lipopolysaccharide (LPS) as an adjuvant to induce autoimmune arthritis. LPS from Escherichia coli was intraperitoneally injected into DBA/1J mice together with the joint cartilage component type II collagen (CII) on day 0. Thereafter, the injection of CII and LPS was continued every 2 weeks up to day 56. The results showed that mice injected with CII plus LPS had signs of arthritis on day 55 and the joint inflammation reached a peak on day 75. Injection of CII or LPS alone induced no arthritis. Histologically, marked oedema of synovium and intense infiltration of inflammatory cells, including neutrophils, were observed 3 days after the onset of joint inflammation. Twenty one days later, there were marked proliferation of synovial tissues with many mononuclear cells and destruction of cartilage. Anti-CII immunoglobulin G (IgG) and IgG2a antibodies were markedly produced in mice injected with CII plus LPS. Pronounced secretion of cytokines, including interleukins-12 and -1beta, interferon-gamma and tumour necrosis factor-alpha, was also observed in these animals. Arthritis was passively transferred into naive syngeneic mice with sera but not with lymphoid cells from mice given CII with LPS. Other types of LPS from Salmonella enteritidis, Salmonella typhimurium and Klebsiella pneumoniae as well as lipid A from E. coli, induced inflammation in joints when administered with CII. Polymixin B sulphate mixed with LPS or lipid A blocked the induction of joint inflammation. These results indicate that LPS appears to play an important role as an adjuvant in the induction of arthritis in which autoimmunity to CII is involved. PMID- 10792510 TI - Gene gun-mediated delivery of an interleukin-12 expression plasmid protects against infections with the intracellular protozoan parasites Leishmania major and Trypanosoma cruzi in mice. AB - An interleukin-12 (IL-12) expression plasmid was transferred, using a gene gun, to mice infected with Leishmania major or Trypanosoma cruzi. Transfer of the IL 12 gene to susceptible BALB/c mice resulted in regression of lesion size and reduced the number of parasites in draining lymph nodes (LN) at the site of L. major infection. Coincident with these protective effects, the T-helper type (Th) response shifted towards Th1, as evaluated by cytokine production in vitro and L. major-specific antibody responses. Protective effects of the IL-12 gene were also observed in T. cruzi infection. Treatment of BALB/c mice infected with T. cruzi enhanced the production of interferon-gamma (IFN-gamma) by spleen cells, while suppressed production of interleukin-10 (IL-10) compared with control mice. Administration of anti-CD4 or anti-CD8 monoclonal antibody (mAb) abolished the protective immunity against T. cruzi infection, and treatment with the IL-12 gene could not restore the resistance in these mice. Mice depleted of natural killer (NK) cells with anti-asialo GM1 also became susceptible to infection, while the resistance was restored when these mice were treated with the IL-12 gene. Thus, target cells for the treatment appear to be CD4+ and CD8+ T cells, which are ordinarily activated by NK cells. These results suggest that the transfer of cytokine genes using a gene gun is an effective method for investigating the roles of cytokines and gene therapy in infectious diseases. PMID- 10792511 TI - Common antigenicity between Japanese cedar (Cryptomeria japonica) pollen and Japanese cypress (Chamaecyparis obtusa) pollen, I. H-2 complex affects cross responsiveness to Cry j 1 and Cha o 1 at the T- and B-cell level in mice. AB - Common antigenicity among two purified Japanese cedar pollen allergens (Cry j 1 and Cry j 2) and one Japanese cypress pollen allergen (Cha o 1) was explored at the T-cell and B-cell level in mice of different H-2 haplotypes. Cry j 2 did not show any common antigenicity with Cry j 1 or Cha o 1. B10.S (H-2S) mice immunized with Cry j 1 or Cha o 1 generated T cells and antibodies reactive to both antigens, indicating the common antigenicity of these antigens. C57BL/6 (H-2b) mice were non-responders to Cry j 1. BALB/c (H-2d) mice immunized with Cry j 1 or Cha o 1 and C57BL/6 mice immunized with Cha o 1 generated T cells that were only reactive with the respective immunogen, but produced antibody reactive to both Cry j 1 and Cha o 1, indicating that Cry j 1 and Cha o 1 share their B-cell epitope but not their T-cell epitope. This finding may provide a clue for the clarification of the T-cell and B-cell epitopes of Cry j 1 and Cha o 1, even though the data are influenced by H-2 complex restriction in mice. Considering that H-2 complex restriction affects cross responsiveness to Cry j 1 and Cha o 1 at the T- and B-cell level in mice, we assessed the possible situation in humans exposed sequentially to Japanese cedar pollen and Japanese cypress pollen. PMID- 10792512 TI - Common antigenicity between Japanese cedar (Cryptomeria japonica) pollen and Japanese cypress (Chamaecyparis obtusa) pollen, II. Determination of the cross reacting T-cell epitope of cry j 1 and cha o 1 in mice. AB - We have previously detected common antigenicity between Cry j 1 and Cha o 1 in B10.S mice. B10.S mice immunized with Cry j 1- or Cha o 1-generated T cells and antibodies reactive to both allergens. In the present study, we investigated the cross-reacting and Cry j 1-specific T-cell epitopes in B10.S mice. Lymph node cells from B10. S mice immunized with Cry j 1 recognized Cry j 1 p111-130, p211 230, and p310-330 as well as Cha o 1 p209-228. The existence of the cross reacting T-cell epitope in Cry j 1 and Cha o 1 was confirmed by the response of newly established p211-230-specific and Cha o 1 p209-228-specific T-cell lines. The minimum peptide sequence (p213-224) of the cross-reacting T-cell epitope was identical in Cry j 1 and Cha o 1. These findings clearly demonstrate that common antigenicity at the T-cell level between Japanese cedar and cypress pollen allergens was caused by the existence of an identitical-cell epitope in Cry j 1 and Cha o 1. PMID- 10792513 TI - Role of F1F0-ATPase in the growth of streptococcus mutans GS5. AB - The role of F1F0-ATPase in Streptococcus mutans GS5 was investigated by isolating a mutant (NTS1) defective in enzyme activity by homologous recombination with a plasmid encoding the 5' terminal fragment of the F1F0-ATPase beta-subunit gene. The ATPase activity of NTS1 membranes was 49% that of GS5 membranes. The lag phase of the growth curve of NTS1 was longer than that for GS5, and the lag phase of GS5 and NTS1 were prolonged by the addition of ionophore gramicidin D; at stationary phase, the turbidity of the NTS1 culture was less than that of the GS5 culture. These results suggest that S. mutans F1F0-ATPase contributes to the generation of a stoichiometric electrogenic gradient effectively in the lag phase. PMID- 10792514 TI - Production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from Kimchi. AB - Lactococcus lactis subsp. lactis A164 was isolated from Kimchi (Korean traditional fermented vegetables). The bacteriocin produced by strain A164 was active against closely related lactic acid bacteria and some food-borne pathogens including Staphylococcus aureus, Listeria monocytogenes and Salmonella typhimurium. The antimicrobial spectrum was nearly identical to that of nisin. Bacteriocin activity was not destroyed by exposure to elevated temperatures at low pH values, but the activity was lost at high pH values. This bacteriocin was inactivated by pronase E and alpha, beta-chymotrypsin, but not by trypsin, pepsin, and alpha-amylase. Cultures of L. lactis subsp. lactis A164 maintained at a constant pH of 6.0 exhibited maximum production of the bacteriocin. It was purified to homogeneity by ammonium sulphate precipitation, sequential ion exchange chromatography, and ultrafiltration. Tricine-SDS-PAGE of purified bacteriocin gave the same molecular weight of 3.5 kDa as that of nisin. The gene encoding this bacteriocin was amplified by PCR with nisin gene-specific primers and sequenced. It showed identical sequences to the nisin gene. These results indicate that bacteriocin produced by Lactococcus lactis A164 is a nisin-like bacteriocin. PMID- 10792515 TI - Characterization of an arylesterase from Lactobacillus helveticus CNRZ32. AB - An esterase gene (estA) was isolated from a previously constructed genomic library of Lactobacillus helveticus CNRZ32. The estA gene consisted of a 558 bp open reading frame encoding a putative peptide of 21.3 kDa. Protein sequence homology searches using BLAST revealed that EstA had low amino acid sequence identity with the serine-dependent arylesterases TesI (24%) and EtpA (26%) from Escherichia coli and Vibrio mimicus, respectively. A recombinant EstA fusion protein containing a C-terminal six-histidine tag was constructed and purified to electrophoretic homogeneity. Characterization of EstA revealed that it was a serine-dependent enzyme having a monomeric Mr of 22.6-25.1 kDa. Optimum temperature, NaCl concentration and pH for EstA activity were determined to be 35 40 degrees C, 3.5% NaCl and 7.5-8.0, respectively. EstA had significant activity under conditions simulating those of ripening cheese (10 degrees C, 4% NaCl, pH 5.1). EstA hydrolysed a variety of ester compounds and preferred those with substituted phenyl alcohol and short-chain fatty acid groups. Site-directed mutagenesis suggested that the S10 and H164 residues were essential for EstA activity. PMID- 10792516 TI - Isolation, properties and behaviour of tyramine-producing lactic acid bacteria from wine. AB - Wines containing high levels of biogenic amines were investigated for the presence of tyramine-producing strains. Two different Lactobacillus brevis (IOEB 9809 and IOEB 9901) able to produce the amine were isolated. None of the isolated strains identified as Oenococcus oeni formed tyramine. In addition, other Lact. brevis and Lact. hilgardii strains from our collection (IOEB) and the American Type Culture Collection (ATCC) were strong tyramine producers. Lactobacillus brevis IOEB 9809 and Lact. hilgardii IOEB 9649 were found to produce tyramine and phenylethylamine simultaneously. The conditions that can influence tyramine formation in wine were evaluated for three strains of Lact. brevis (IOEB 9809 and IOEB 9901) and Lact. hilgardii (IOEB 9649). Tyrosine was the major factor affecting tyramine formation and was enhanced by the presence of sugars, mainly glucose. Tyrosine decarboxylase (TDC) activity greatly depended on the presence of the precursor, which suggested that tyrosine induced the TDC system. These results indicate that Lactobacillus could be the lactic acid bacteria responsible for tyramine production in wine. PMID- 10792517 TI - Limitation of adhesion and growth of Listeria monocytogenes on stainless steel surfaces by Staphylococcus sciuri biofilms. AB - The adhesion and subsequent development of Listeria monocytogenes on stainless steel was studied in the absence and in the presence of a Staphylococcus sciuri biofilm. In the three growth media studied, the percentage of adherent cells was reduced to nearly the same extent by the presence of 1-day biofilms of Staph. sciuri for the two strains of L. monocytogenes studied. One-day biofilms of Staph. sciuri exhibited the same exopolysaccharide content per square centimetre, although they colonized from 3.5 to 35% of the stainless steel depending on the growth media. This suggests that extracellular substances rather than cell-to cell interactions were involved in the decreased adhesion. After 3 days of culture, Staphylococcus biofilms prevented the adherent L. monocytogenes population from increasing within the biofilm, leading to an average logarithmic cfu difference of 0.9-2.7 between the pure and mixed culture. A competition for nutrients by Staph. sciuri was observed in one of the three media. A role for extracellular polysaccharides produced by the Staphylococcus biofilm in preventing the adhesion of L. monocytogenes and in modifying the balance existing between its planktonic and biofilm phase is hypothesized. A higher proportion of L. monocytogenes cells was observed in the planktonic phase in mixed cultures, suggesting that the extracellular substances produced by Staph sciuri biofilms and involved in the decreased adhesion of L. monocytogenes could modify the balance existing between planktonic and biofilm populations. In addition, co cultures of L. monocytogenes and Staph. sciuri in broth showed competition for nutrients for Staph. sciuri in one of the three media. PMID- 10792518 TI - Effect of modified atmosphere packaging and water activity on growth of Eurotium amstelodami, E. chevalieri and E. herbariorum on a sponge cake analogue. AB - The combined effects of water activity (aw), oxygen (O2) and carbon dioxide (CO2) levels on growth variables of three species of Eurotium (two isolates of each) on a sponge cake analogue were studied. The use of a multivariate statistical method (PLS) for the analysis of data showed that in general, the fungi had the same behaviour, according to their growth responses, to changes in the three tested factors. However, the maximal growth of E. herbariorum was somewhat more affected by the O2 level at lower CO2 levels. On the other hand, CO2, aw and the interaction between CO2 and O2 were found to be the most significant factors describing growth variables. In addition, the model found in this study had a good predictive power. There was a positive correlation between the CO2 concentration in the bags and the lag phase, and a negative correlation between the growth rate and maximum mycelial growth. In general, no isolates were able to growth when CO2 concentrations were higher than 60% under anaerobic conditions. At lower values of CO2, aw and O2 influenced growth variables. Low levels of O2 (0.02-0. 5%) did not affect the growth variables studied when levels of CO2 in the bags were high. However, when the CO2 concentration decreased, the different O2 levels had an influence on the growth variables, i. e., a small difference in the concentration of O2 can allow fungi to grow. PMID- 10792519 TI - Prevalence, characterization and growth of Bacillus cereus in commercial cooked chilled foods containing vegetables. AB - In cooked-chilled and pasteurized vegetable products, initial numbers of Bacillus cereus were below 10 cfu g-1. Before the appearance of spoilage, numbers reached 6-8 log cfu g-1 at 20 degrees C and 4-6 log cfu g-1 at 10 degrees C. Bacillus cereus was not detected in samples stored at 4 degrees C. Ten percent of strains isolated from the products were able to grow at 5 degrees C and 63% at 10 degrees C. Bacillus cereus strains unable to degrade starch, a feature linked to the production of emetic toxin, did not grow at 10 degrees C and had a higher heat resistance at 90 degrees C. Using immunochemical assays, enterotoxin was detected in the culture supernatant fluid of 97.5% of the strains. All culture supernatant fluids were cytotoxic but important variations in the level of activity were found. Psychrotrophic isolates of B. cereus were unable to grow in courgette broth at 7 degrees C whereas they grew in a rich laboratory medium. At 10 degrees C, these isolates grew in both media but lag time in courgette broth was 20-fold longer than in the rich laboratory medium. PMID- 10792520 TI - Thermal inactivation of Listeria monocytogenes and Yersinia enterocolitica in minced beef under laboratory conditions and in sous-vide prepared minced and solid beef cooked in a commercial retort. AB - D-values were obtained for Listeria monocytogenes and Yersinia enterocolitica at 50, 55 and 60 degrees C in vacuum-packed minced beef samples heated in a laboratory water-bath. The experiment was repeated using vacutainers, which allowed heating of the beef to the desired temperature before inoculation. D values of between 0.15 and 36.1 min were obtained for L. monocytogenes. Pre heating the beef samples significantly affected (P < 0.05) the D60 value only. D values for Y. enterocolitica ranged from 0.55 to 21.2 min and all the D-values were significantly different (P < 0.05) after pre-heating. In general, the D values obtained for core inoculated solid beef samples were significantly higher (P < 0.05) than those generated in minced beef when heated in a Barriquand Steriflow commercial retort. PMID- 10792521 TI - Influence of environmental factors on virus detection by RT-PCR and cell culture. AB - The effects of clay, humic acid, u.v. light and shellfish tissue residues on the detection of poliovirus type 2 from environmental samples by culture and RT-PCR were investigated. RT-PCR showed 10-100 times greater sensitivity for PV2 detection in the absence of sample contaminants than did culture by plaque assay in BGM cell monolayers. Bentonite clay (100-1000 mg l-1) and shellfish tissue residues reduced virus detection by plaque assay, but the effect of bentonite was mitigated by simple elution procedures. Bentonite clay, humic acid (5-150 mg l-1) and mussel tissue reduced virus detection by RT-PCR by between 1 and 8 logs, although this was mitigated in part by elution and Sephadex filtration of extracts. Sephadex filtration of samples reduced culturable PV2 by 32-50%. Exposure of PV2 in water to u.v. light reduced culturability of PV2 but not detection by RT-PCR. This study demonstrates that virus detection in environmental samples is strongly influenced by naturally occurring substances and disinfection approaches. The accuracy of results of viral analyses of this nature should be carefully scrutinized with respect to sample constituents. PMID- 10792522 TI - Biodegradation of didecyldimethylammonium chloride by Pseudomonas fluorescens TN4 isolated from activated sludge. AB - Bacteria that degrade didecyldimethylammonium chloride (DDAC) were isolated from activated sludge from a municipal sewage treatment plant by enrichment culture with DDAC as a sole carbon source. One of the isolates, Pseudomonas fluorescens TN4, degraded DDAC to produce decyldimethylamine and subsequently, dimethylamine, as the intermediates. The TN4 strain also assimilated the other quaternary ammonium compounds (QACs), alkyltrimethyl- and alkylbenzyldimethyl-ammonium salts, but not alkylpyridinium salts. TN4 was highly resistant to these QACs and degraded them by an N-dealkylation process. These data mean that there are some QAC-resistant and QAC-degrading bacteria such as TN4 in the environment. PMID- 10792523 TI - Characterization of Bacillus anthracis strains used for vaccination. AB - Three Bacillus anthracis strains, formerly used as anti-anthrax vaccine strains in Argentina, were characterized from genetic and pathogenic perspectives. Southern blotting and PCR with pXO1 and pXO2 probes and primers, as well as pathogenicity and protection tests in guinea pigs and mice, were performed. Two of the B. anthracis strains contained both pXO1 and pXO2 plasmids, as did the fully virulent strains, while the third was a Sterne-type strain (pXO1+, pXO2-). The three strains were, however, markedly less pathogenic than a wild-type virulent strain. The methodology applied here may be used to characterize other B. anthracis strains. PMID- 10792525 TI - A kinetic model for energy spilling-associated product formation in substrate sufficient continuous culture. AB - It has been demonstrated that excess substrate can cause uncoupling between anabolism and catabolism, which leads to energy spilling. However, the Luedeking Piret equation for product formation does not account for the energy spilling associated product formation due to substrate excess. Based on the growth yield and energy uncoupling models proposed earlier, a kinetic model describing energy spilling-associated product formation in relation to residual substrate concentration was developed for substrate-sufficient continuous culture and was further verified with literature data. The parameters in the proposed model are well defined and have their own physical meanings. From this model, the specific productivity of unit energy spilling-associated substrate consumption, and the maximum product yield coefficient, can be determined. Results show that the majority of energy spilling-associated substrate consumption was converted to carbon dioxide and less than 6% was fluxed into the metabolites, while it was found that the maximum product yield coefficients varied markedly under different nutrient limitations. The results from this research can be used to develop the optimized bioprocess for maximizing valuable product formation. PMID- 10792524 TI - Optimization of high-molecular-weight polycyclic aromatic hydrocarbons' degradation in a two-liquid-phase bioreactor. AB - A microbial consortium degrading the high-molecular-weight polycyclic aromatic hydrocarbons (HMW PAHs) pyrene, chrysene, benzo[a]pyrene and perylene in a two liquid-phase reactor was studied. The highest PAH-degrading activity was observed with silicone oil as the water-immiscible phase; 2,2,4,4,6,8, 8 heptamethylnonane, paraffin oil, hexadecane and corn oil were much less, or not efficient in improving PAH degradation by the consortium. Addition of surfactants (Triton X-100, Witconol SN70, Brij 35 and rhamnolipids) or Inipol EAP22 did not promote PAH biodegradation. Rhamnolipids had an inhibitory effect. Addition of salicylate, benzoate, 1-hydroxy-2-naphtoic acid or catechol did not increase the PAH-degrading activity of the consortium, but the addition of low-molecular weight (LMW) PAHs such as naphthalene and phenanthrene did. In these conditions, the degradation rates were 27 mg l-1 d-1 for pyrene, 8.9 mg l-1 d-1 for chrysene, 1.8 mg l-1 d-1 for benzo[a]pyrene and 0.37 mg l-1 d-1 for perylene. Micro organisms from the interface were slightly more effective in degrading PAHs than those from the aqueous phase. PMID- 10792526 TI - The physiology and pathogenicity of Mycobacterium tuberculosis grown under controlled conditions in a defined medium. AB - A chemically-defined culture medium was developed which supported batch growth of Mycobacterium tuberculosis, strain H37Rv, at a minimum doubling time of 14.7 h. This medium also facilitated chemostat culture of M. tuberculosis at a constant doubling time of 24 h. Chemostat growth was optimized at a dissolved oxygen tension of 20% (v/v) and 0.2% (v/v) Tween-80. Chemostat cultures were dispersed suspensions of single bacilli (1.5-3 microm long), or small aggregates, at a mean density of log10 8.3 cfu ml-1. A limited number of amino acids was utilized (alanine, asparagine, aspartate and serine were depleted by >50%; glycine, arginine, isoleucine, leucine and phenylalanine, by approximately 40%). Chemostat grown cells were pathogenic in aerosol-infected guinea pigs, producing disseminated infection similar to that caused by plate-grown cells. Cells from chemostat culture were significantly more invasive for J774A.1 mouse macrophages than agar- or batch-grown cells. This study demonstrates the suitability of chemostat culture for the growth of pathogenic mycobacteria in a defined physiological state with potential applications for the controlled production of mycobacterial components for therapeutic and vaccine applications. PMID- 10792527 TI - Inactivation of Bacillus subtilis spores on packaging surfaces by u. v. excimer laser irradiation. AB - Ultraviolet (u.v.) laser irradiation has been used to inactivate Bacillus subtilis spores deposited on to planar aluminium- and polyethylene-coated packaging surfaces. Kill kinetics were found to be diphasic, with an initial rapid inactivation phase followed by tailing. Although no definitive evidence was obtained, it is thought that spores located within packaging crevices/pores were primarily responsible for the observed tailing. Surviving spores were also found on the unexposed underside of cards and, to a lesser extent, within clumps. The log count reduction in B. subtilis was dependent on spore loading and total u.v. dose. In comparison, packaging surface composition, fluence (2-18 Jm-2) and frequency (40-150 Hz) had only a negligible effect. By irradiating boards carrying 106 spores, with a dose of 11.5 J cm-2, a log count reduction >5 was obtained. The mode of spore inactivation was primarily through DNA disruption. This was confirmed by the high sensitivity of spores lacking protective, small, acid-soluble proteins, in addition to the high frequency of auxotrophic and asporogenous mutations found amongst survivors. PMID- 10792528 TI - Effect of sausage ingredients and additives on the production of enterocin A and B by Enterococcus faecium CTC492. Optimization of in vitro production and anti listerial effect in dry fermented sausages. AB - Enterocin A and B in Enterococcus faecium CTC492 were co-induced by the different factors assayed in this study (r = 0.93) and followed primary metabolic kinetics. Enterocin production was significantly inhibited by sausage ingredients and additives, with the exception of nitrate. The addition of sodium chloride and pepper decreased production 16-fold. The temperature and pH influenced enterocin production, with optima between 25 and 35 degrees C, and from 6.0 to 7.5 of initial pH. The maximum activity was achieved, under favourable growth conditions, with MRS supplemented with sucrose (2%) plus glucose (0.25%) and Tween-80 (1%). MRS concentration, NaCl plus pepper addition, absence of Tween-80 in the growth medium, incubation at 45 degrees C and an initial pH under 5.5 were detrimental to bacteriocin production. Stress conditions did not favour enterocin production. Desadsorption was Tween-dependent. Enterocin A activity in the crude extracts stored at -80 degrees C was better preserved than enterocin B (when tested against their specific indicator strain), but anti-listerial activity remained intact. Applied as anti-listerial additives in dry fermented sausages, enterocins significantly diminished Listeria counts by 1. 13 log (P < 0.001), while Enterococcus faecium CTC492 added as starter culture did not significantly reduce Listeria counts (P > 0. 1) compared with the standard starter culture (Bac ). Enterocins A and B could be considered as extra biopreservative hurdles for listeria prevention in dry fermented sausages. PMID- 10792529 TI - Iron requirement of Lactobacillus spp. in completely chemically defined growth media. AB - A completely chemically-defined growth medium, containing guanine, thymine, cytidine, 2'-deoxyadenosine and 2'-deoxyuridine as DNA precursors, was developed for Lactobacillus johnsonii, on the basis of statistically designed techniques suitable for other lactobacilli. Particular focus was given to the nucleotide composition of different defined media, and to the specific nucleotide requirements of Lact. johnsonii. Most of the lactobacilli tested grew in a medium containing five free bases, four ribonucleosides or five deoxyribonucleosides. Adenine and guanine were replaceable by inosine. The requirement for thymine and cytosine was satisfied with uracil. The presence of inosine and uracil was identified as being essential for the growth of different Lactobacillus species, displaying their inability to synthesize purines and pyrimidines de novo. Defined recipes with different nucleotide composition were used to investigate iron requirements of lactobacilli. Only marginal differences in growth were observed in iron-depleted media supplemented with five free bases, four ribonucleosides or five deoxyribonucleosides; iron depletion had a greater effect on growth when inosine and uracil were supplied as the only nucleotide sources. The results suggest that iron plays a role in the pyrimidine and purine metabolism of lactobacilli. Lactobacillus spp., particularly Lact. johnsonii, require iron under particular environmental conditions with limited or specific nucleotide sources. PMID- 10792530 TI - The Pseudomonas group as an indicator of potential regrowth in water distribution systems. AB - The regrowth of micro-organisms in the water distribution system is assisted by the presence of biodegradable organic matter (BDOC). The low concentration of available organic carbon in this type of water favours the growth of bacteria belonging to the Pseudomonadaceae family, as this group can grow better at low concentrations of substrate than other bacteria also present in the distribution system. Although the genus Aeromonas has already been adopted as an indicator of this potential regrowth, members of the Pseudomonadaceae have not yet been proposed as indicators of potential bacterial regrowth in the water distribution system. The results are presented of a year-long study of the Barcelona distribution system in which the presence of Pseudomonas and Aeromonas was analysed and the validity of these micro-organisms as indicators of potential regrowth in the distribution system was assessed. It seems that, at least in the drinking waters of the Barcelona area, Pseudomonas is a better indicator of potential bacterial regrowth than Aeromonas. PMID- 10792531 TI - Use of the MIDI-FAME technique to characterize groundwater communities. AB - Fatty acid methyl ester (FAME) profiles were identified directly from groundwater microbial communities concentrated on and extracted with polycarbonate filters. The sensitivity of this direct extraction method was determined using pure cultures of Acinetobacter junii, Pseudomonas putida and Stenotrophomonas maltophilia. A minimum concentration of 107 cells filter-1 was required to identify the predominant fatty acids from each culture. However, at least 3.7 x 109 cells filter-1 were required to obtain fatty acid profiles that matched the signature profiles for pure cultures in a commercial database. While several saturated fatty acids (i.e. 14 : 0, 16 : 0, 18 : 0) were extracted from the polycarbonate filters, they were readily subtracted from microbial fatty acid profiles and did not interfere with the characterization of pure cultures or environmental samples. For the environmental samples, 3 l of groundwater from the Savannah River Site, Aiken, SC, (USA) contained sufficient biomass for direct extraction. A comparative analysis of FAME groundwater profiles demonstrated a qualitative difference among communities sampled from spatially discrete locations, while a groundwater well that was sampled at two time points showed strong similarities over time. Concentration of microbial biomass on polycarbonate filters coupled with the MIDI-FAME extraction of both biomass and filter was a useful technique to characterize microbial communities from groundwater. PMID- 10792532 TI - Modulation of Salmonella infection by the lectins of Canavalia ensiformis (Con A) and Galanthus nivalis (GNA) in a rat model in vivo. AB - The plant lectins, Concanavalin A (Con A) and Galanthus nivalis agglutinin (GNA) have been prefed to rats for 3 d pre- and 6 d postinfection with Salmonella typhimurium S986 or Salm. enteritidis 857. Con A significantly increased numbers of Salm. typhimurium S986 in the large intestine and in faeces, and severely impaired growth of the rats, more severely than is the case of infection with Salmonella typhimurium alone. Con A had much less effect on rats infected with Salm. enteritidis 857 only showing a significant increase in numbers in the colon, accompanied by intermittent increases of Salmonella in the faeces during the study. GNA significantly reduced pathogen numbers in the lower part of the small bowel and the large intestine of rats infected with Salm. typhimurium S986 and significantly improved rat growth. GNA had little effect on infection by Salm. enteritidis 857 with slight decreases in Salmonella numbers in the small intestine and large intestine and transient increases in the faeces. PMID- 10792533 TI - Virulence factors of Escherichia coli O157 and other Shiga toxin-producing E. coli. PMID- 10792534 TI - The stress response is repressed during fermentation in brewery strains of yeast. AB - Yeast cells encounter a variety of environmental stresses during brewing and must respond to ensure cell survival. Cells can respond to stress by inducing a Heat Shock Response in which heat shock proteins (Hsps) are synthesized. In laboratory strains of Saccharomyces cerevisiae, the heat shock protein, Hsp104, plays a major role in the acquisition of tolerance to a variety of stresses such as heat, ethanol and sodium arsenite, and as such acts as an excellent stress indicator. The induction of Hsp104 in bottom-and top-fermenting brewery strains was examined when grown under laboratory and industrial fermentation conditions, and it was found that each brewing strain exhibits its own unique pattern of Hsp104 expression. During industrial fermentations, brewery strains are capable of mounting a stress response at the early stages of fermentation. However, as the fermentation proceeds, the response is repressed. The results suggest that conditions experienced in industrial brewing prevent the activation of the stress response. This study increases our understanding of alterations in gene expression patterns during the brewing process, and yields information that will aid in the definition of best practice in yeast management. PMID- 10792535 TI - Dose-response relationships. A model for describing interactions, and its application to the combined effect of nisin and lactic acid on Leuconostoc mesenteroides. AB - As well as producing bacteriocins, many lactic bacteria produce other potentially toxic compounds or growth inhibitors, especially lactic acid, which may interfere in the assays commonly used to quantify these peptides. A systematic set of modifications is proposed which, when applied to the logistical equation, enable it to describe the combined (but not additive) effects of two or more active principles. The general model thus derived is applied to the interaction of nisin and lactic acid on Leuconostoc mesenteroides. PMID- 10792536 TI - Degradation of 4-nitrocatechol by Burkholderia cepacia: a plasmid-encoded novel pathway. AB - Pseudomonas cepacia RKJ200 (now described as Burkholderia cepacia) has been shown to utilize p-nitrophenol (PNP) as sole carbon and energy source. The present work demonstrates that RKJ200 utilizes 4-nitrocatechol (NC) as the sole source of carbon, nitrogen and energy, and is degraded with concomitant release of nitrite ions. Several lines of evidence, including thin layer chromatography, gas chromatography, 1H-nuclear magnetic resonance, gas chromatography-mass spectrometry, spectral analyses and quantification of intermediates by high performance liquid chromatography, have shown that NC is degraded via 1,2, 4 benzenetriol (BT) and hydroquinone (HQ) formation. Studies carried out on a PNP- derivative and a PNP+ transconjugant also demonstrate that the genes for the NC degradative pathway reside on the plasmid present in RKJ200; the same plasmid had earlier been shown to encode genes for PNP degradation, which is also degraded via HQ formation. It is likely, therefore, that the same sets of genes encode the further metabolism of HQ in NC and PNP degradation. PMID- 10792537 TI - Molecular characterization of the Thermomonospora curvata aglA gene encoding a thermotolerant alpha-1,4-glucosidase. AB - The cloning, sequencing and structural characterization of a gene encoding a thermostable alpha-1,4-glucosidase from Thermomonospora curvata is described. DNA sequence analysis revealed four open reading frames designated aglA, aglR, aglE and aglF. The aglA gene encodes a thermostable alpha-1,4-glucosidase from T. curvata and is situated between two genes, aglR and aglE. Genes aglA, aglE and aglF are transcribed in the same direction, while aglR is transcribed in the opposite direction. By comparing the amino acid sequence of the alpha-1,4 glucosidase from T. curvata with other alpha-glucanases, it appears that the enzyme is a member of the alpha-amylase family. The proteins of this family have an (alpha/beta)8 barrel super secondary structure. The topology of the alpha-1,4 glucosidase was predicted by computer-assisted analysis. The topology of the secondary structures of the alpha-1,4-glucosidase resembles the structure of barley alpha-amylase, but the primary structure resembles most closely the oligo 1,6-glucosidase from Bacillus cereus. Putative catalytic residues (D221, E281 and D343) and calcium binding residues (N116, E179, D191, H224 or G225) are proposed. PMID- 10792538 TI - Susceptibility testing: accurate and reproducible minimum inhibitory concentration (MIC) and non-inhibitory concentration (NIC) values. AB - Measuring the minimum inhibitory concentration (MIC) of a substance by current methods is straightforward, whereas obtaining useful comparative information from the tests can be more difficult. A simple technique and a method of data analysis are reported which give the experimentalist more useful information from susceptibility testing. This method makes use of a 100-well microtitre plate and the analysis uses all the growth information, obtained by turbidometry, from each and every well of the microtitre plate. A modified Gompertz function is used to fit the data, from which a more exact value can be obtained for the MIC. The technique also showed that at certain concentrations of inhibitor, there was no effect on growth relative to a control well (zero inhibitor). Above a threshold value, which has been termed the non-inhibitory concentration or NIC, growth becomes limiting until it reaches the MIC, where no growth relative to the control is observed. PMID- 10792539 TI - Phospholipid analogues of Porphyromonas gingivalis. AB - Porphyromonas has lipids containing hydroxy acids and C16:0 and iso-C15:0 major monocarboxylic acids among others. Nothing is known of its individual phospholipid molecular species. The aim of this study was to determine molecular weights and putative identities of individual phospholipid molecular species extracted from Porphyromonas gingivalis (seven strains), P. asaccharolytica (one strain) and P. endodontalis (two strains). Cultures on Blood-Fastidious Anaerobe Agar were harvested, washed and freeze-dried. Phospholipids were extracted and separated by fast atom bombardment mass spectrometry (FAB MS) in negative-ion mode. Phospholipid classes were also separated by thin layer chromatography (TLC). The major anions in the range m/z 209-299 were consistent with the presence of the C13: 0, C15: 0, C16: 0 and C18: 3 mono-carboxylate anions. Major polar lipid anion peaks in the range m/z 618-961 were consistent with the presence of molecular species of phosphatidylethanolamine, phosphatidylglycerol and with unidentified lipid analogues. Porphyromonas gingivalis differed from comparison strains of other species by having major anions with m/z 932, 946 and 960. Unusually, a feline strain of P. gingivalis had a major peak of m/z 736. Selected anions were studied by tandem FAB MS which revealed that peaks with m/z 653 and 946 did not correspond to commonly occurring classes of polar lipids. They were however, glycerophosphates. It is concluded that the polar lipid analogue profiles obtained with Porphyromonas are quite different from those of the genera Prevotella and Bacteroides but reveal heterogeneity within P. gingivalis. PMID- 10792540 TI - Production and characterization of a thermostable chitinase from a new alkalophilic Bacillus sp. BG-11. AB - An alkalophilic, environmental micro-organism, Bacillus sp. BG-11, has been isolated and characterized. It produced 76 U ml-1 of chitinase in liquid batch fermentation after 72 h of incubation at 50 degrees C using chitin-enriched medium. The molecular weight of purified chitinase was estimated to be 41 kDa by SDS-PAGE. The pH and temperature optima of chitinase immobilized on chitosan and calcium alginate were 8.5 and 50 degrees C, respectively, which were same as that of free enzyme. The pH and thermostability of immobilized chitinase were enhanced significantly. The chitinase immobilized on chitosan was stable between pH 5.0 and 10.0, and the half-life of chitosan-immobilized enzyme at 70, 80 and 90 degrees C was 90, 70 and 60 min, respectively. The end-products formed during the enzyme-substrate reaction were identified by 13C-NMR, and N-acetyl-D-glucosamine was found to be the major end-product. GlcNAc (GlcNAc)2 and (GlcNAc)3 inhibited the chitinase activity by 32, 25 and 18%, respectively, at a concentration of 10 mmol l-1. The shelf-life of chitinase (retained 100% activity) at 4 degrees C was 8 weeks in the presence of either sodium azide (100 microgram ml-1), sodium metabisulphite (0.1% w/v) or KCl (15% w/v). The enzyme was resistant to the action of proteases and allosamidin. PMID- 10792541 TI - Rapid fluorescence assessment of intracellular pH as a viability indicator of Clavibacter michiganensis subsp. michiganensis. AB - The viability of Clavibacter michiganensis subsp. michiganensis (Cmm) was determined by measuring the intracellular pH (pHin) as a viability parameter. This was based on the observation that growth of Cmm was inhibited at pH 5.5 and below. Therefore, viable cells should maintain their pHin above this pH value. The pHin of Cmm was determined using the fluorescent probe 5(and 6-) carboxyfluorescein succinimidyl ester (cFSE). The pHin of Cmm cells exposed to acid treatments was determined using fluorescence spectrofluorometry, and for cells exposed to elevated temperatures, the pHin was determined using fluorescence spectrofluorometry and flow cytometry (FCM). A good correlation was found between the presence of a pH gradient and the number of colony-forming units (cfu) observed in plate counts. However, with the spectrofluorometry technique, the analysis is based on the whole cell population and the detection sensitivity of this technique is rather low, i.e., cell numbers of at least 107 cfu ml-1 are needed for the analysis. Using FCM, heat-treated and non-treated Cmm cells could be distinguished based on the absence and presence of a pH gradient, respectively. The major advantage of FCM is its high sensitivity, allowing analysis of microbial populations even at low numbers, i.e., 102-103 cfu ml-1. PMID- 10792542 TI - Biochemical properties of Streptococcus macedonicus strains isolated from Greek Kasseri cheese. AB - A total of 32 Streptococcus macedonicus strains, isolated from Greek Kasseri cheese, were screened for biochemical properties of technological importance in milk fermentation processing, such as acid production, proteolytic and lipolytic activity, citrate metabolism, exopolysaccharide production, antimicrobial activity and biogenic amines production. All strains were found to be moderate acidifiers in milk. Only four strains could hydrolyse milk casein, while 11 strains showed lipolytic activity against tributyrin. Using amino acid derivatives of 4-nitroaniline as substrates, the highest peptidase activities were determined against phenylalanine- and glycine-proline-4-nitroanilide. Using fatty acid derivatives of 4-nitrophenol, it was shown that all strains exhibited esterase activities up to caprylate, with highest values against butyrate and caproate. Only one showed activity up to palmitate; this was also the most active strain against tributyrin. Five of the 32 strains could metabolize citrate but none of them produced exopolysaccharides. Nine strains displayed antimicrobial activity towards Clostridium tyrobutyricum, while no antimicrobial activity was detected against Listeria innocua and Propionibacterium freudenreichii subsp. shermanii. Finally, none was able to decarboxylize ornithine, histidine or lysine, and only four strains produced tyramine from tyrosine. PMID- 10792543 TI - A fluorescently-labelled r-RNA targeted oligonucleotide probe for the in situ detection of G-bacteria of the genus Amaricoccus in activated sludge. AB - A fluorescently-labelled r-RNAtargeted oligonucleotide probe specific for members of the genus Amaricoccus, which includes one group of the Gram-negative G Bacteria seen in activated sludge systems, is described. These organisms, previously 'identified' on their distinctive morphology of cocci in tetrads, have been associated with poor performance of biological nutrient removal (EBNR) plants, by out-competing the polyphosphate accumulating bacteria. Methods of sample preparation for probing activated sludge are detailed, and preliminary surveys of 46 plants, using this probe, show that G-Bacteria belonging to the genus Amaricoccus are seen not only in large numbers in EBNR systems but also in conventional plants. The presence of single cells of this organism was common, emphasizing the dangers of relying on morphology and cell arrangement to identify these bacteria. PMID- 10792545 TI - Evaluation of the environmental impact of microbial aerosols generated by wastewater treatment plants utilizing different aeration systems. AB - Using three sampler devices (SAS, Andersen Six-Stages and All Glass Impinger), the environmental impact of bacterial and fungal aerosols generated by municipal wastewater treatment plants operating with different methods of sludge oxygenation were evaluated. The highest microbial concentrations were recovered above the tanks (2247 cfu m-3) and in downwind positions (1425 cfu m-3), where a linear correlation (P < 0.05) was found between the quantity of sewage treated and the entities of microbial aerosol dispersion. Moreover, an exponential increase (P < 0.05) in the bacteria recovered from the air occurred at increasing times of treatment. However, after long-term plant operation, high bacterial and fungal concentrations were found in almost all of the sites around the plant. Coliforms, enterococci, Escherichia coli and staphylococci were almost always recovered in downwind positions. Considerable fractions (20-40%) of sampled bacteria were able to penetrate the final stages of the Andersen apparatus and thus, are likely to be able to penetrate the lungs. The plant operating with a fine bubble diffused air system instead was found to generate rather low concentrations of bacteria and fungi; moreover, staphylococci and indicator micro organisms were almost absent. Finally, salmonellae, Shigella, Pseudomonas aeruginosa and Aeromonas spp. were not detected in either of the plants. The results indicate a remarkable dispersion of airborne bacteria and fungi from tanks in which oxygen is supplied via a mechanical agitation of sludge, and suggest the need to convert them to diffused aeration systems which pose a lesser hazard for human health. PMID- 10792544 TI - Effects of heat stress on the antimicrobial drug resistance of Escherichia coli of the intestinal flora of swine. AB - The effects of heat stress on the antimicrobial drug resistance of Escherichia coli of the intestinal tract of swine were studied in animals from a farm that had not been supplementing antimicrobials in feed for the past 10 years. In one study, 10 finisher hogs were heat stressed (34 degrees C) for 24 h. Antimicrobial resistance levels after stress were significantly higher (P < 0.05) when compared with pre-stress levels for amikacin, ampicillin, cephalothin, neomycin and tetracycline from faecal samples. This high level of resistance persisted to slaughter that occurred at 10 days post-stress for most of the antimicrobials mentioned. In a second study, samples of different sections of the gastrointestinal tract were collected after heat stress and compared with control, non-stressed animals. Results indicated that E. coli which colonized the ileum and caecum had a higher level of resistance to ampicillin and tetracycline than the E. coli which colonized the colon and rectum. When animals were exposed to heat stress, resistance to ampicillin and tetracycline of E. coli in the lower digestive tract increased (P < 0.05) to a level similar to that observed in the ileum and caecum. Based on these findings, an investigation was made to test the hypothesis that (a) an increase in intestinal motility increases shedding of resistant E. coli and (b) heat stress induces a reduction in intestinal transit time in swine. For each study, two groups of three, randomly selected finisher hogs each were formed (treated and control groups). In study (a), induction of increased motility and peristalsis was obtained using an intramuscular injection of the cholinergic drug neostigmine methylsulphate. Escherichia coli isolates were obtained from the ileum, caecum, colon and rectum after animals were slaughtered. A higher level of ampicillin-resistant E. coli was found in the caecum (40%) than in other segments of the intestinal tract. In treated animals, level of resistance increased for organisms from the colon and rectum. Similar results were obtained for tetracycline resistance. In study (b), intestinal transit time was measured using chromium-EDTA as a marker. Swine were euthanized and samples were collected throughout the intestinal tract (duodenum to rectum) 8 h after administration of the marker to control and heat-stressed animals. Results indicated a reduced transit time for the stressed group. These findings corroborate the initial hypothesis that an outflow of resistant organisms moves from the upper tract (ileum and caecum) to the lower tract (colon and rectum). PMID- 10792546 TI - Analysis of denitrification by Pseudomonas stutzeri under nutrient-limited conditions using membrane inlet mass spectrometry. AB - Membrane inlet mass spectrometry (MIMS) was used to investigate denitrification by Pseudomonas stutzeri in a static lake water column. Continuous real-time measurement of gases enabled the dynamics of the process to be investigated. Concentrations of 17 mmol l-1 nitrate and 10 mmol l-1 nitrite were identified as optimal for denitrification under nutrient-limited conditions (i.e., produced the highest concentrations of N2). Available carbon was the major rate-limiting factor in lake water when nitrate or nitrite was present. No stratification of the process with depth was observed, and aerobic denitrification was apparent under all the conditions employed. The rate of denitrification was dependent on cell concentration, and possible limitations of the usefulness of MIMS under environmentally modelled conditions were identified for environments containing low numbers of bacteria. PMID- 10792548 TI - Improved detection of Listeria monocytogenes in soft mould-ripened cheese. AB - In comparison with standard methods, enrichment in half-Fraser broth for 24 h at 30 degrees C, followed by plating out onto Listeria monocytogenes blood agar (LMBA) and PALCAM medium combined with an additional streak proved to be the most rapid and specific method for the detection of indigenous L. monocytogenes populations from soft mould-ripened cheese. This procedure, with a high sensitivity (93%) and a low detection limit (1-10 cfu 25 g-1), provided negative and presumptive positive results within 2-3 d. Differences between LMBA, PALCAM and Oxford medium turned out to be highly significant (at 99% significance level); plating on LMBA after standard enrichment protocols giving the best overall results. An improvement in detection was also obtained by modifying the confirmation procedure. A loopful of culture (an additional streak) from PALCAM or Oxford medium was streaked on non-selective medium in addition to streaking only separate colonies as specified in the standards. PMID- 10792547 TI - Development of a genetically modified bacteriophage for use in tracing sources of pollution. AB - Bacteriophage are frequently used as biotracers to identify the source of water pollutants. Genetic manipulation of bacteriophage M13mp18 has been used to enhance this technique by creating a library in which each recombinant bacteriophage genome contains a unique identification sequence. Techniques that identify a recombinant bacteriophage by the presence of the identification sequence, including polymerase chain reaction, restriction site polymorphism and plaque hybridization, have been developed. Recombinant bacteriophage can be used to test a large number of suspected sources simultaneously. The identification sequence also eliminates confusion with natural bacteriophage present in water samples. The performance of the modified bacteriophage and the techniques were assessed in simulated field trials on a restricted site carried out under a consent for environmental release of a genetically modified organism. The techniques were also field tested at sites in northwest England using wild-type M13 bacteriophage. PMID- 10792549 TI - Isolation and characterization of a new bacteriocin from Lactobacillus gasseri KT7. AB - A bacteriocin-producing Lactobacillus gasseri strain, KT7, was isolated from infant faeces. The supernatant fluid showed inhibitory activity not only against some lactic acid bacteria but also, against some pathogenic and food-spoilage species, including Clostridium, Listeria and Enterococcus. An antimicrobial peptide designated gassericin KT7 was isolated from Lactobacillus gasseri KT7. It was purified to homogeneity by a single four-step procedure: a crude supernatant fluid obtained from early stationary-phase culture in MRS medium was subjected to ammonium sulphate fractionation, CM-Sephadex cation-exchange chromatography, Phenyl-Sepharose hydrophobic chromatography and reverse-phase HPLC chromatography. Gassericin KT7 was sensitive to proteolytic enzymes, resistant to heat, active over a wide range of pH, and migrated as a 4.5-5.0 kDa peptide on SDS-PAGE. The bacteriocin was produced constitutively during exponential growth. It was bactericidal to sensitive cells and the bactericidal effect was not produced by cell lysis. The amino acid composition of the bacteriocin was determined and no modified amino acid was found among the residues identified. PMID- 10792550 TI - Effect of steroidal saponin from Yucca schidigera extract on ruminal microbes. AB - The effects of steroidal saponins (SAP) isolated from Yucca schidigera extract on ruminal bacteria and fungi were investigated in pure culture studies. Prevotella bryantii, Ruminobacter amylophilus, Selenomonas ruminantium and Streptococcus bovis were cultured through ten 24-h transfers in ruminal fluid medium containing 0 or 25 microg SAP ml-1 (measured as smilagenin equivalents). The four strains, each non-exposed or pre-exposed to SAP, were then inoculated into medium containing 0 or 250 microgram smilagenin equivalents ml-1 and 24-h growth curves were determined. The cellulolytic ruminal bacteria Ruminococcus flavefaciens, Fibrobacter succinogenes and Rc. albus were cultured for 72 h on Whatman no. 1 filter paper in medium containing 0, 9, 90 or 180 microgram SAP ml-1 for the determination of filter paper digestion and endoglucanase activity. The ruminal bacteria differed in their responses to SAP. Steroidal saponins in the medium reduced the growth of Strep. bovis (P < 0.01 at 2, 3, 4, 5, 6 and 8 h), P. bryantii (P < 0.05 at 4, 5, 6, 8, 10 and 24 h) and Rb. amylophilus (P < 0.05 at 14 and 24 h), but the growth of S. ruminantium was enhanced (P < 0.05) at 10, 14 and 24 h. The growth curves of all four non-cellulolytic species were similar (P > 0.05) between pre-exposed and non-exposed cultures and the concentrations of total SAP and soluble (deglycosylated) SAP in the liquid fraction were unchanged (P > 0.05) over time. Steroidal saponins inhibited the digestion of filter paper by all three cellulolytic bacteria, but F. succinogenes was less (P < 0.05) sensitive to SAP and more (P < 0. 05) effective at deglycosylating SAP than were Rc. flavefaciens or Rc. albus. Transmission electron microscopy revealed that SAP altered the cell walls of the SAP-inhibited non-cellulolytic bacteria. The ruminal fungi, Neocallimastix frontalis and Piromyces rhizinflata, were cultured on filter paper in medium containing 0, 0. 45, 2.25 or 4.5 microgram SAP ml-1. Filter paper digestion by both fungi was completely inhibited by 2.25 microgram SAP ml-1. Steroidal saponins from Y. schidigera inhibit cellulolytic ruminal bacteria and fungi, but their effects on amylolytic bacteria are species dependent and similar to the effects of ionophores. As such, SAP may be useful in nutritional applications targeting starch-digesting ruminal micro-organisms. PMID- 10792552 TI - Disinfection kinetics: a new hypothesis and model for the tailing of log survivor/time curves. AB - A new hypothesis for the understanding of chemical disinfection, which we have termed the Intrinsic Quenching hypothesis, is presented. This mechanistic treatment of disinfection kinetics is based on the hypothesis that the biocide concentration may not be in vast excess over the microbes, as is normally assumed. A mathematical model was developed and found to be useful in describing the observed kinetics of several disinfectants. The model suggested that the reason for the observation of non-linear, log-survivor curves was due to the ability of the microbes, in clean, soil-free conditions, to intrinsically quench the bulk concentration of biocide. PMID- 10792551 TI - Interaction between Aeromonas veronii and epithelial cells of spotted sand bass (Paralabrax maculatofasciatus) in culture. AB - An in vitro fish model to study the interaction between Aeromonas veronii and skin, gill and intestinal epithelial cells was developed using primary cultures of mucosal cells (isolated from healthy organisms). Primary cultures were exposed to Aeromonas veronii strain A186 isolated from a patient with severe gastrointestinal disease. Microbial adherence was assessed by a spectrophotometric evaluation of an enzyme-linked, biotin-streptavidin Aer. veronii cell-adhesion assay to confluent monolayers of epithelial cells on 96 well tissue culture plates. The three primary-culture cells are susceptible to Aer. veronii attachment, with the greatest binding affinity found in gills, and to a lesser extent, in skin and intestine epithelial cells. Aer. veronii adherence was dependent on bacterial load and incubation time. The effect of glycoconjugates on Aer. veronii adhesion was investigated by pre-incubating Aer. veronii cells with monosaccharides, sialic acid-rich glycoproteins and sulphated polysaccharides. In addition, the participation of a 48-kDa Aer. veronii lectin (MCBP - mucosal constituents binding protein), with affinity for mucosal constituents, was evaluated as a putative adhesion factor of Aer. veronii to the mucosal epithelial cells of spotted sand bass by pre-incubating bacterial cells with rabbit polyclonal antibodies to Aer. veronii MCBP. Our study shows that primary-culture fish mucosal cells provide a suitable model for the study of the interactions between Aer. veronii and epithelial cells of the fish mucosa, and to study putative virulence factors of fish pathogens. PMID- 10792553 TI - Multiple chromosomes in Burkholderia cepacia and B. gladioli and their distribution in clinical and environmental strains of B. cepacia. AB - Burkholderia cepacia is found in soils and waters, it can be used in biocontrol and bioremediation but is also a human pathogen. It is not yet clear what differentiates pathogenic from non-pathogenic strains of the organism. In this study the multiple replicon structure was investigated in 28 strains of B. cepacia by pulsed field gel electrophoresis. All strains examined, whether of clinical, environmental or plant pathogenic origin, were found to have two, three or four large (> 500 kbp) replicons. Many strains also contained small replicons. Clinical strains were more likely to have three or four large replicons than non clinical strains. Multiple replicon structure was also demonstrated in B. gladioli and Alcaligenes eutrophus. PMID- 10792554 TI - Prospects for progress in diagnostic imaging. AB - New fast-imaging MRI systems designed specifically for cardiac magnetic resonance enable new applications of noninvasive vascular imaging. The use of functional MRI and diffusion tensor imaging to map brain function and structure offers a new dimension to an understanding of the human condition. Clinical applications of functional MRI will influence many specialties including surgery, education, and rehabilitation. Functional imaging has the potential to visualize the regional concentration of specific proteins. This imaging at the level of molecules may be possible by use of a contrast material whose signal is changed by local enzymatic activity. The three-dimensional digital data collected in modern imaging techniques allow for virtual endoscopy in the respiratory, alimentary, and cardiovascular systems. Virtual endoscopy may replace many of the more invasive diagnostic methods in the near future. The measurement of clinical decision making through observer performance studies better informs both the physician and the patient on how to improve upon the quality of clinical practice. These prospects for progress reinforce diagnostic imaging as a cornerstone in medical informatics. The history of creating images used in medicine reveals the invention of diagnostic tools which may provide new information but premature use can result in improper application of a poorly understood technology. Research into the use of new technology may be as important as the technology itself in improving the human condition. PMID- 10792555 TI - Adverse endothelial function and the insulin resistance syndrome. AB - Type 2 diabetes is characterized by impaired endothelial dependent vasodilatation which may contribute to the high prevalence of vascular disease in such patients. Although hyperglycaemia, dyslipidaemia and hypertension can all independently cause a similar defect, recent data suggest that endothelial dysfunction may be intrinsic to the insulin resistance syndrome that commonly precedes type 2 diabetes. Such abnormalities in endothelial function could represent the impact of subclinical disturbance of metabolism or alternatively the presence of a common cellular defect that influences both nitric oxide bioavailability and insulin mediated glucose disposal. Resolution of this puzzle is likely to lead to important advances in our knowledge and ultimately treatment of vascular disease. PMID- 10792556 TI - The EWA (estrogen in women with atherosclerosis) study: a randomized study of the use of hormone replacement therapy in women with angiographically verified coronary artery disease. Characteristics of the study population. Effects on lipids and lipoproteins. AB - OBJECTIVES: To evaluate the effects of hormone replacement therapy (HRT) on lipids and lipoproteins in postmenopausal women with coronary artery disease. SETTING: In this single-centre, controlled and randomized study taking place in a tertiary referral clinic, patients were examined at baseline, and after 3 and 12 months. All analyses were performed examiner-blind. SUBJECTS: Postmenopausal women (n = 118) with angiographically verified coronary artery disease were recruited consecutively from patients referred for investigational procedures due to coronary artery disease. INTERVENTIONS: The women were randomized to HRT, i.e. transdermal application of continuous 17-beta oestradiol with cyclic medroxyprogesterone actetate tablets every 3rd month for 14 days, or to a control group. MAIN OUTCOMES: Effects on lipids and lipoproteins. RESULTS: After 3 months of unopposed oestradiol, triglycerides decreased significantly compared to the control group (P = 0.006). Sequential administration of medroxyprogesterone caused a decrease in HDL cholesterol (P = 0.01), concomitantly with a decrease in ApoA1 lipoproteins (P = 0.007). No other changes in lipids or lipoproteins were observed. After 12 months of therapy, no significant differences were observed between the two groups in lipid or lipoprotein levels. Concomitant statin treatment did not alter the main findings. CONCLUSIONS: In postmenopausal women with established coronary artery disease in whom the majority is treated with statins, no additional effect of HRT on lipids or lipoproteins could be observed except for a transient decrease in triglycerides in the initial unopposed oestradiol phase. No deleterious effect could be observed during medroxyprogesterone administration except for a small transient decrease in HDL cholesterol and ApoA1 lipoproteins. PMID- 10792558 TI - Ten-year mortality amongst patients with a very small or unconfirmed acute myocardial infarction in relation to clinical history, metabolic screening and signs of myocardial ischaemia. AB - AIM: To evaluate the long-term prognosis amongst patients with a very small or unconfirmed acute myocardial infarction (AMI) in relation to clinical history, metabolic screening and signs of myocardial ischaemia at exercise test. METHODS: Patients with a very small or unconfirmed AMI, aged < 76 years, were selected and given a clinical evaluation, metabolic screening and checked for ischaemia at an exercise test 4 weeks after admittance. The 10-year mortality was related to age, sex, clinical history, body weight, serum (S) cholesterol, S-triglycerides, S gammaglutamyltranspeptidase (GT), S-glucose and various indices of myocardial ischaemia at exercise test. RESULTS: In all, 714 patients participated in the evaluation. The median age was 63 years and 33% were women. The overall 10-year mortality was 33%. In univariate analysis, the following factors appeared as risk indicators for death: age (P < 0.0001), a history of previous AMI (P < 0.0001), angina pectoris (P < 0.001), diabetes mellitus (P < 0.0001), congestive heart failure (P < 0.0001), smoking (P = 0.030), S-triglycerides (P < 0.0001), S-gamma GT (P < 0. 0001) and S-glucose (P < 0.0001). In multivariate analysis, the following remained as independent risk indicators for death: age (P < 0.0001), S gamma GT (P < 0.0001), previous AMI (P < 0.0001), smoking (P < 0.0001) and S glucose (P = 0.010). CONCLUSION: Amongst patients with a very small or a unconfirmed AMI, factors reflecting their clinical history, including age, a history of AMI and current smoking, as well as factors reflecting their metabolic status, including S-gamma GT and S-glucose, were important predictors for the long-term outcome. PMID- 10792557 TI - Hyperhomocysteinaemia is not associated with isolated crural arterial occlusive disease: The Hoorn Study. AB - OBJECTIVES: Hyperhomocysteinaemia is an independent risk factor for peripheral arterial disease (PAD). The localization of peripheral arterial disease is clinically relevant, because proximal (aortoiliac and femoropopliteal) disease is associated with a particularly poor overall prognosis, whereas isolated distal (i.e. crural) disease is associated with a better overall prognosis. The aim of the study was to investigate whether the strength of the association between hyperhomocysteinaemia and peripheral arterial disease differs according to the localization of the anatomical obstruction. DESIGN: Fasting serum total homocysteine (tHcy) was measured in an age-, sex- and glucose-tolerance stratified random sample (n = 631) of a 50- to 75-year-old general Caucasian population. History of a peripheral arterial reconstruction was recorded. Aortoiliac, femoropopliteal and crural arterial obstructions were registered by means of Doppler flow velocity curves. RESULTS: The median serum tHcy level was 12.2 micromol L-1 (interquartile range: 10.0-15.3) in men and 10.7 micromol L-1 (interquartile range: 9.0-13.3) in women. The prevalences of aortoiliac, femoropopliteal and crural obstructions were 2.1%, 2.7% and 11.9%, respectively. After adjustment for age, sex, systolic blood pressure, current smoking, serum cholesterol and diabetes mellitus, the odds ratios (95% confidence interval) per 5 micromol L-1 tHcy increment were 1.41 (1.05-1.89) for aortoiliac, 1.03 (0. 70 1.52) for femoropopliteal and 0.82 (0.59-1.15) for crural obstructions. Finally, diabetes mellitus, HbA1c and current smoking were significantly associated with crural and femoropopliteal disease, whereas systolic blood pressure was significantly associated with aortoiliac obstructions. CONCLUSIONS: The present study indicates that hyperhomocysteinaemia is associated with aortoiliac but not with isolated crural arterial occlusive disease. PMID- 10792559 TI - Mechanisms behind gender differences in circulating leptin levels. AB - OBJECTIVES: To investigate gender differences in circulating leptin levels and adipose tissue production of leptin. DESIGN SETTING AND SUBJECTS: Thirty-two men and 63 women with a large interindividual variation in body mass index (BMI), but otherwise healthy, were investigated after an overnight fast. Body fat (bioimpedance), abdominal subcutaneous adipose tissue secretion of leptin in vitro and serum leptin were determined. RESULTS: Although there was no gender difference in mean BMI or fat cell size, mean percentage body fat was 49 in women and 36 in men (P < 0.001). At each level of BMI, serum leptin levels were about two times higher in women than in men (P < 0.001). Adipose tissue secretion rate of leptin in men was two-thirds of that in women (P < 0.05). The gender differences in body fat content, serum leptin and leptin secretion were observed in obese (BMI > 27 kg m-2) as well as non-obese subjects. Serum leptin levels (P < 0.001) and leptin secretion rate (P < 0.01) correlated positively with body fat content in either sex. However, the gender differences in serum leptin (P < 0.001) and leptin secretion rate (P < 0.01) remained statistically significantly different even when the values were adjusted for body fat. CONCLUSION: The gender difference in circulating leptin concentrations can be due to at least two different mechanisms. A higher proportion of adipose tissue and increased production rate of leptin per unit mass of adipose tissue might explain why women have higher circulating leptin levels than men. PMID- 10792561 TI - Activity of cardiopulmonary baroreceptors, peripheral resistance and cutaneous microcirculation in patients with peripheral obstructive arterial disease. AB - OBJECTIVES: To assess the effects of cardiopulmonary baroreceptors on the haemodynamics of the humeral and common carotid arteries in patients suffering from peripheral artery disease (PAD) and to discover whether the stimulation of these receptors modifies the cutaneous microcirculation in the forearm. DESIGN: We studied a group of patients suffering from peripheral artery disease and two groups as controls. SETTING: Patients were examined at the ambulatory for Vascular Research, Division of Vascular Medicine and Rehabilitation, University of Verona. PATIENTS: We studied 15 patients with peripheral artery disease of the lower limbs at Fontaine stage II (group C), 10 free of arterial pathologies (group B) and 10 young people (group A). MAIN OUTCOME MEASURES: We subjected the patients to passive elevation of the legs and the trunk in a horizontal position with pressure monitoring and measurement of the calibre and flow in the brachial and common carotid arteries using a colourDoppler ultrasound. We also studied the cutaneous microcirculation with laserDoppler flowmetry. RESULTS: During the test, arterial pressure and cardiac frequency remained constant in group A, systolic pressure values showed a slight, but statistically significant increase in group B, whilst the increase in systolic pressure values at this stage was marked in group C. Diastolic pressure values and cardiac frequency remained unchanged in all groups. The calibre of the humeral artery increased in the control groups. Carotid resistance was unchanged in the three groups. Humeral resistance during the test decreased in the two control groups whilst it increased in group C. The number of perfusion units felt in the control groups; no variations in group C. CONCLUSIONS: Our study demonstrates, in patients with peripheral artery disease, a reduction in the activity of the cardiopulmonary baroreceptors with an increase in the humeral resistance during the test and impairment of the mechanisms of cutaneous microcirculatory vasoregulation in the forearm. PMID- 10792560 TI - Hormone replacement therapy in healthy postmenopausal women. Effects on intraplatelet cyclic guanosine monophosphate, plasma endothelin-1 and neopterin. AB - OBJECTIVES: To evaluate beneficial effects of postmenopausal hormone replacement therapy (HRT) on endothelial function, measured as intraplatelet cyclic guanosine monophosphate (cGMP, mediator of nitric oxide), cyclic adenosine monophosphate (cAMP, mediator of prostacyclin) and plasma endothelin-1 (ET-1), and on monocyte activation, measured as plasma neopterin. DESIGN: Part 1: double-blind randomized trial for 3 months; part 2: open study for 9 months. SETTING: The study was performed at the Department of Endocrinology, University Hospital, Malmo, Sweden. SUBJECTS: Fifty-one postmenopausal women participated in part 1 and 46 in part 2. Inclusion criteria included a history of amenorrhoea for at least 6 months before the study and body mass index >/= 24 kg m-2. INTERVENTION: Randomization for either placebo (n = 24) or HRT (n = 27). HRT was given as 2 mg oestradiol valerate for the first 3 months with the addition of 10 mg medroxyprogesterone for 10 days every third month thereafter. MEASUREMENTS: Performed at baseline and after 3 and 12 months of the study. RESULTS: In the HRT group, intraplatelet cGMP increased from 0.56 (0.35-0.94) to 0.61 (0.42-3. 40) and 0.65 (0.43-1.08) pmol (109 platelets)-1 after 3 and 12 months, respectively (P = 0.01), whereas plasma ET-1 decreased from 3.2 (1.1-6.8) to 2.0 (0.8-5.1) and 1.8 (0.4-15.4) pg mL-1 (P < 0. 001). Intraplatelet cAMP and plasma neopterin were unchanged. When smokers (n = 15) and non-smokers (n = 12) in the HRT group were analysed separately, significant effects were seen only amongst smokers. The control group showed unchanged levels of cGMP, cAMP, ET-1 and neopterin. CONCLUSIONS: These data suggest beneficial effects of HRT on endothelial function which may account for anti-atherogenic effects of HRT in postmenopausal women, especially in smokers. No effects of HRT were seen upon monocyte activation. PMID- 10792562 TI - Association of coronary heart disease with age-adjusted aortocoronary calcification in patients with familial hypercholesterolaemia. AB - OBJECTIVES: Existing algorithms of risk of coronary heart disease (CHD) do not pertain to patients with familial hypercholesterolaemia (FH), whose arteries have been exposed to hypercholesterolaemia since birth. We studied a cohort of FH patients to compare four diagnostic models of CHD: traditional risk factors of CHD (age, sex, cholesterol, hypertension, smoking and body mass index), cholesterol year score, and aortic as well as coronary calcium measured by spiral computed tomography (CT). SUBJECTS: We invited 88 individuals with molecularly defined FH of whom 80 (91%) decided to participate. RESULTS: Analysis of receiver operating characteristic curves showed that the age-adjusted coronary calcium score was more strongly associated with clinical manifestations of CHD than were traditional risk factors (P < 0.002), cholesterol year score (P << 0.0001), and the age-adjusted aortic calcium score (P < 0.0004). CONCLUSIONS: Age-adjusted coronary calcium score shows promise as an indicator of CHD in FH patients. PMID- 10792563 TI - Advanced glycation end product in familial amyloidotic polyneuropathy (FAP). AB - OBJECTIVES: Advanced glycation end products (AGE) are present in amyloid deposits in beta2-microglobulin amyloidosis, and it has been postulated that glycation of beta2-microglobulin may be involved in fibril formation. The aim of this paper was to ascertain whether AGE occur in amyloid deposits in familial amyloidotic polyneuropathy (FAP). SETTING: Department of Medicine, Umea University Hospital and First Department of Internal Medicine, Kumamoto University School of Medicine. DESIGN: The presence of AGE was sought immunohistochemically and biochemically in amyloid-rich tissues from patients with FAP. SUBJECTS: Biopsy specimens from nine patients and 10 controls were used for the immunohistochemical analysis. For amyloid preparation, vitreous samples from three FAP patients were used. RESULTS: Immunohistochemical studies using a polyclonal anti-AGE antibody revealed positive immunoreactivity in intestinal materials, but the pattern of reactivity was unevenly distributed; it was often present in the border of amyloid deposits, or surrounding them. Non-amyloid associated immunoreactivity was also observed in a few regions of the specimens, although the AGE-positive structures were situated in areas containing amyloid deposits. Western blotting of purified amyloid from the vitreous body of FAP patients revealed a significant association of AGE with amyloid fibrils. CONCLUSION: The immunoreactivity for the AGE antibody suggests that AGE may be involved in fibril formation in FAP. PMID- 10792564 TI - Primary prophylaxis for individuals with severe haemophilia: how many hospital visits could treatment prevent? AB - OBJECTIVES: To assess how many hospital visits primary prophylaxis with clotting factor could prevent. DESIGN: The potential for reducing hospital visits was assessed by comparing rates of in-patient, Out-patient and day-case visits per patient-year for individuals with severe (< 1 IU dL-1) haemophilia who had never received primary prophylaxis with attendance rates for individuals with mild/moderate (1-50 IU dL-1) haemophilia. Hospital attendance data were collected retrospectively for the period 1988-97 inclusive for individuals who were aged 18 years or over. SETTING: Data were obtained on patients who were registered at the Katharine Dormandy Haemophilia Centre (KDHC), London, UK. OUTCOME MEASURES: In patient stays, Out-patient and day-case visits. RESULTS: Individuals with mild/moderate haemophilia were 45% (31-56), 36% (30-41) and 70% (68-73) less likely to have required in-patient, Out-patient and day-case visits than were individuals with severe haemophilia. HIV serostatus and age were also shown to be significant and independent predictors of the rate of Out-patient and day-case visits, but not the rate of in-patient stays. CONCLUSION: These results suggest that primary prophylaxis for individuals with severe haemophilia could significantly reduce the demand for in-patient stays, and Out-patient and day case visits. PMID- 10792565 TI - Mortality, risk indicators of death, mode of death and symptoms of angina pectoris during 5 years after coronary artery bypass grafting in men and women. AB - AIM: To describe mortality, risk indicators of death, mode of death and symptoms of angina pectoris during 5 years after coronary artery bypass grafting in women and men. SAMPLE: All patients in western Sweden who underwent coronary artery bypass grafting without concomitant valve surgery and without previously performed coronary artery bypass grafting between June 1988 and June 1991. RESULTS: In all, 2000 patients participated in the evaluation, 381 (19%) of whom were women. Compared to men, who had a 5-year mortality of 13.3%, women had a relative risk of death of 1.4 (95% CI 1.0-1.8; P = 0.03). Renal dysfunction interacted significantly (P = 0.048) with gender, in that the differences were more marked in patients without renal dysfunction. When adjusting for differences at baseline, the relative risk of death amongst women was 1.0 (95% CL 0.7-1.3). Compared to men, women had an increased risk of in-hospital death and death associated with stroke. However, amongst the patients who died, the place and mode of death appeared to be similar in women and men. Amongst survivors after 5 years, women had more symptoms of angina pectoris than men. CONCLUSION: During 5 years after coronary artery bypass grafting, women had an increased mortality compared to men; renal dysfunction seemed to interact with female gender regarding mortality. Women had a higher risk of in-hospital death and death associated with stroke. However, the adjusted relative risk of death during 5 years was equal in women and men. Amongst survivors, women suffered more from angina pectoris than men. PMID- 10792566 TI - The marker of cobalamin deficiency, plasma methylmalonic acid, correlates to plasma creatinine. AB - OBJECTIVE: To examine the relationship between the two diagnostic tests, plasma methylmalonic acid and plasma cobalamins, and their association with plasma creatinine, age and sex. DESIGN: Cross-sectional study of simultaneous laboratory measurements. SETTING: County of Aarhus, Denmark. SUBJECTS: Records on 1689 patients who had their first plasma methylmalonic acid measurement during 1995 and 1996, and who had a simultaneous measurement of plasma cobalamins. Plasma creatinine values measured within a week of measurements of plasma methylmalonic acid and plasma cobalamins were available for 1255 of the patients. MAIN OUTCOME MEASURES: Predictors of variation in plasma methylmalonic acid; plasma cobalamins, plasma creatinine, age and sex. RESULTS: Plasma methylmalonic acid was positively correlated with plasma creatinine, even for plasma creatinine within the normal range. These associations remained in a multiple regression analysis. For plasma cobalamins below 200 pmol L-1, there was a strong negative correlation between plasma methylmalonic acid and plasma cobalamins, whilst the association was weak for higher plasma cobalamin levels. Plasma methylmalonic acid increased and plasma cobalamins decreased with age. CONCLUSIONS: The strong correlation between plasma methylmalonic acid and plasma creatinine suggests that plasma creatinine - also within the normal range - must be taken into consideration when interpreting plasma methylmalonic acid. PMID- 10792568 TI - Neurobiology of short stature and prognosis of coronary heart disease in women. PMID- 10792567 TI - Disease-related syncope. Analysis of a community-based hospital registry. AB - Syncope is characterized by sudden and transient loss of consciousness that follows a reduction or interruption of cerebral blood flow. The present study was designed to assess the prevalence of disease-related syncope in a wide sample of in-patients admitted for different diseases. A total of 16 809 patients (age range 18-99 years) were recruited from three hospitals in Florence in 1998. The community-based registry was reviewed to identify all patients suffering from complaints associated with syncope. Each disease-related syncope was matched with the number of patients suffering from that disease. Furthermore, each disease was expressed as a percentage of total cases included in the study. The odds ratio was calculated to determine the index of significant correlation between syncope and occasional diseases. Total syncopes were 775 (prevalence 4.46%), vasovagal syncopes were 336 (1.9% of total sample and 44% of total syncopes), and the disease-related syncopes were 439 (56% of total syncopes). We found a significant association between syncope and orthostatic hypotension, complete heart block, chronic cerebral disease, migraine, acute gastrointestinal haemorrhages and aortic stenosis. Furthermore, we found a significant association with acute gastrointestinal haemorrhage, which has not been described previously. Significant relationships emerged from our data which yield a new insight into the association between syncope and a wide range of systemic diseases. PMID- 10792569 TI - Brooke-Spiegler syndrome locus assigned to 16q12-q13. PMID- 10792570 TI - Desmosome assembly and keratin network formation after Ca2+/serum induction and UVB radiation in Hailey-Hailey keratinocytes. PMID- 10792571 TI - A homozygous missense mutation in the cytoplasmic tail of beta4 integrin, G931D, that disrupts hemidesmosome assembly and underlies Non-Herlitz junctional epidermolysis bullosa without pyloric atresia? PMID- 10792572 TI - Long-term culture of murine epidermal keratinocytes. PMID- 10792573 TI - A novel point mutation of the FALDH gene in a Japanese family with Sjogren Larsson syndrome. PMID- 10792574 TI - Stress, hormones and your brain. PMID- 10792575 TI - Cocaine and amphetamine-regulated-transcript peptide mediation of leptin stimulatory effect on the rat gonadotropin-releasing hormone pulse generator in vitro. AB - Pulsatile gonadotropin-releasing hormone (GnRH) secretion was studied in vitro using explants of the retrochiasmatic hypothalamus from prepubertal male and female rats. Leptin caused a dose-dependent reduction of the GnRH interpulse interval in both sexes. We studied the effects of cocaine- and amphetamine regulated transcript (CART) since this peptide was shown recently to mediate the anorectic effects of leptin in the hypothalamus. CART caused a reduction of the GnRH interpulse interval. This effect was prevented using an anti-CART antiserum which could partially overcome leptin stimulatory effects as well. Using hypothalamic explants from Zucker rats homozygous for the leptin receptor mutation ( fa/fa), GnRH pulse frequency was not affected by leptin, while a significant acceleration was caused by the CART-peptide. In conclusion, leptin involves the hypothalamic CART-peptide to stimulate the prepubertal GnRH pulse generator in vitro. PMID- 10792576 TI - Differential regulation of neuropeptide Y mRNA expression in the arcuate nucleus and locus coeruleus by stress and antidepressants. AB - In rats, circulating corticosterone and insulin are involved in regulation of the hypothalamic neuropeptide Y (NPY) system, which in turn, is involved in regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Since the HPA axis and stress responsivity is altered in diseases such as depression, we investigated interactions between the effects of stress and antidepressant drug treatment on arcuate nucleus and locus coeruleus NPY mRNA expressions using in situ hybridization histochemistry. After acute (2 h) and repeated immobilization (2 h daily, for 14 days), plasma concentrations of corticosterone increased, and those of insulin decreased. The expression of NPY mRNA was significantly increased in the arcuate nucleus, but was unchanged in the locus coeruleus following acute and repeated immobilization. Adrenalectomized rats with systemic corticosterone replacement (ADX+CORT), whose corticosterone concentration was maintained at approximately 50-100 ng/ml during repeated stress, showed a decrease in plasma insulin and an increase in arcuate nucleus NPY mRNA similar to that observed in sham rats, suggesting that changes in NPY mRNA levels are more closely tied to circulating insulin than to circulating corticosterone. In contrast, locus coeruleus NPY mRNA expressions in ADX+CORT rats were significantly higher than those in sham rats after repeated stress. Desmethylimipramine (DMI) treatment for 24 days did not affect basal plasma concentrations of corticosterone or insulin, or arcuate nucleus NPY mRNA expressions, but significantly decreased basal levels of locus coeruleus NPY mRNA compared to saline-treated rats. After repeated immobilization (2 h daily, for 4 days), DMI significantly reduced the stress-induced rise in locus coeruleus NPY mRNA levels, but potentiated the stress-induced rise in arcuate nucleus NPY mRNA expression. These results demonstrate that: (1) the increase in arcuate nucleus NPY mRNA expressions in stressed rats closely follows the decrease in plasma concentrations of insulin; (2) increases in NPY mRNA expressions occur in the absence of changes in plasma corticosterone; and (3) desipramine treatment potentiated the effect of stress on arcuate nucleus NPY mRNA expressions, but blocked the repeated stress-induced increase in locus coeruleus NPY mRNA expressions. Thus, NPY mRNA expression in the arcuate nucleus and the locus coeruleus is sensitive to the effects of stress and to the antidepressant drug desipramine, but the arcuate nucleus NPY system is regulated by different mechanisms than the locus coeruleus NPY system. The results provide further evidence for the importance of circulating insulin in the regulation of the arcuate nucleus NPY system. PMID- 10792577 TI - Calbindin-D28k immunoreactivity is a marker for a subdivision of the sexually dimorphic nucleus of the preoptic area of the rat: developmental profile and gonadal steroid modulation. AB - Calbindin-D28k (calbindin) is a 28 kilodalton calcium binding protein which potentially plays a role in neuroprotection. We report here the normal development and gonadal steroid modulation of a sexually dimorphic group of calbindin immunoreactive cells within the sexually dimorphic nucleus of the preoptic area (SDN) which we call the calbindin-immunoreactive SDN or CALB-SDN. Beginning on PN2, a faintly immunoreactive CALB-SDN is present, however, the volume is not sexually dimorphic. On PN4, the staining of the CALB-SDN appears more robust but the volume is still not sexually dimorphic. By PN8 and extending through PN12 and PN26, the latest age analysed, the volume of the CALB-SDN is larger in males by two- to fourfold. Cresyl violet counterstain reveals a similar developmental profile of the SDN as well as clusters of darkly staining calbindin immunonegative cells which lie around the CALB-SDN. Castration of males on PN0 decreases the volume of the CALB-SDN by PN12 and administration on the day of birth and PN1 of either testosterone propionate or oestradiol benzoate, but not dihydrotestosterone propionate to females increases the volume of the CALB-SDN by PN12. By demonstrating the sexual dimorphism and gonadal steroid modulation of the CALB-SDN, we hereby establish that calbindin is a specific marker of a subdivision of the SDN and can be used as such in future studies. PMID- 10792578 TI - Peripheral or central administration of motilin suppresses LH release in female rats: a novel role for motilin. AB - Motilin is secreted in a clear episodic pattern during fasting or during the interdigestive phase, but feeding promptly stops this secretory pattern, and plasma concentrations of motilin decrease. We have previously determined that fasting markedly suppresses pulsatile luteinizing hormone (LH) secretion in female rats in the presence of oestrogen. In the present study, we wished to learn if motilin may mediate the fasting-induced suppression of LH secretion by determining the effects of motilin administration on LH release and on food intake. Intravenous (i.v.) injection of motilin (37 nmol/rat) suppressed LH release and significantly decreased mean LH concentrations both in ovariectomized (OVX) and oestradiol-implanted ovariectomized (OVX+E2) rats. Food intake was significantly increased by i.v. motilin injection in OVX rats, but not in OVX+E2 rats. It is likely that motilin inhibits LH release via inhibition of the gonadotrophin-releasing hormone (GnRH)-releasing mechanism at the hypothalamic level, because motilin (3.7 nmol/rat) also suppressed LH secretion when centrally administered, and because LH release in i.v. motilin-treated rats increased in response to exogenous GnRH. These results suggest that motilin may be a peripheral signal for the suppression of LH secretion through central sensors. PMID- 10792579 TI - Oxytocin and vasopressin neurones in principal and accessory hypothalamic magnocellular structures express Fos-immunoreactivity in response to acute glucose deprivation. AB - Neurohypophyseal secretion of oxytocin and vasopressin is elevated in response to decreased systemic glucose availability. In these studies, dual-label immunocytochemistry was used to identify hypothalamic neuropeptidergic magnocellular neurones that are transcriptionally activated in response to glucose substrate imbalance. Two h after i.p. injection of the glucose antimetabolite, 2-deoxy-D-glucose (2DG), or the vehicle, saline, groups of adult male rats were anaesthetized by i.p. injection with sodium pentobarbital and killed by transcardial perfusion. Sections (25 microm) through anterior and tuberal levels of the hypothalamus were processed for nuclear Fos- and cytoplasmic neuropeptide immunoreactivity (-ir). A high proportion of oxytocin-ir neurones in the supraoptic, paraventricular, and adjunct structures, including the anterior commissural, periventricular magnocellular, posterior perifornical, recurrent supraoptic, medial forebrain, and circular nuclei, were colabelled for nuclear Fos-ir following administration of 2DG. Large numbers of vasopressin neurones in the supraoptic, circular, posterior perifornical, and medial forebrain nuclei, and posterior magnocellular division and posterior subnucleus of the paraventricular nucleus were also immunostained for Fos in rats injected with the antimetabolite. These results show that decreased glucose metabolism is a stimulus for activation of the Fos stimulus-transcription cascade within oxytocin-and vasopressin-immunopositive neurones in several hypothalamic loci, findings that reflect activation of the Fos-stimulus transcription cascade within large proportions of these cell populations during this metabolic challenge. These data suggest that both peripheral hormonal and central modulatory functions of these neuropeptidergic neurones may be influenced by cellular glucose availability. PMID- 10792580 TI - Insulin-like growth factor I in the anterior pituitary of the clawed frog Xenopus laevis: immunocytochemical and autoradiographic indication for a paracrine action and corelease with prolactin. AB - Insulin-like growth factor I (IGF-I) and its receptor are present in human and rat anterior pituitary. However, few data exist on the potential presence of IGF I or its receptor in the non-mammalian pituitary and the cellular sites of IGF-I production have not been identified in any species. Thus, we investigated the anterior pituitary of the clawed frog Xenopus laevis which is widely used to study growth and differentiation. The study was performed with antisera against mammalian insulin-like growth factor I (IGF-I), prolactin (PRL) and growth hormone (GH) using immunohistochemical and immunocytochemical techniques. IGF-I binding was determined by in-vitro receptor autoradiography. The PRL-and GH immunoreactive cells exhibited distinct distribution patterns. Neither at the light nor the electron microscopical level any colocalization of PRL-and GH immunoreactivities was apparent. The PRL-immunoreactive cells exhibited round granules of medium electron density (mean diameter: 312 nm) and the GH immunoreactive cells spherical granules of medium electron density (mean diameter: 165 nm). By the use of serial semithin sections IGF-I-immunoreactivity was exclusively located in PRL-immunoreactive cells. At the ultrastructural level, IGF-I-immunoreactivity was confined to the secretory granules in coexistence with PRL-immunoreactivity using the double labelling immunogold technique. Specific IGF-I binding sites were localized throughout the pituitary. The results provide evidence for a concomitant release of PRL and IGF-I and suggest autocrine/paracrine actions of IGF-I in the anterior pituitary. PMID- 10792581 TI - Effect of interleukin-1beta on gonadotropin-releasing hormone (GnRH) and GnRH receptor gene expression in castrated male rats. AB - Increasing evidence suggests that interleukin-1beta (IL-1beta) regulates luteinizing hormone (LH) release primarily through modulation of the gonadotropin releasing hormone (GnRH) neuronal activity. This study was undertaken to elucidate the effect of IL-1beta on GnRH as well as GnRH receptor (GnRHR) gene expression in the preoptic area. IL-1beta (100 ng/rat) or saline was administered into the lateral ventricle of castrated rats. RNA samples were isolated from micropunches of the preoptic area and mediobasal hypothalamus from individual brain slices and GnRH mRNA levels in the preoptic area and GnRHR mRNA levels in the mediobasal hypothalamus were determined by competitive reverse transcription polymerase chain reaction (RT-PCR) protocols. Serum LH concentrations were decreased from 1 h to 3 h after IL-1beta treatment, but rebounded at 5 h, while serum concentrations of follicle-stimulating hormone (FSH) and prolactin were not altered. There were no significant changes in GnRH mRNA levels from the micropunched preoptic area, while GnRHR mRNA levels from the preoptic area and mediobasal hypothalamus micropunch samples, but not in the anterior pituitary, showed a pattern similar to the serum LH profile following i.c.v. administration of IL-1beta. We then examined the effect of IL-1beta on the translational efficiency of the GnRH mRNA. After the separation and fractionation of polyribosome-associated cytoplasmic RNA from the hypothalamic fragments containing the preoptic area-anterior hypothalamic area of control (saline treated) and IL-1beta-treated group 3 h after administration, GnRH transcript levels were examined from the each fraction. IL-1beta decreased the translational efficiency of the transcribed GnRH mRNA. These results clearly demonstrate that central administration of IL-1beta suppresses the translational activity of GnRH mRNA. Moreover, GnRHR may play an important role in the modulation of GnRH neuronal activity through GnRHR-expressing neurones (or glia) in the hypothalamus. PMID- 10792582 TI - The active phase-related increase in corticosterone and aggression are linked. AB - Recently we demonstrated that corticosterone exerts an acute facilitatory effect on aggression in male rats. Corticosterone production reaches a maximum at the onset of the dark period, while male rats are more aggressive in the dark. Here we present evidence demonstrating that the corticosterone increase at the beginning of the dark period is causally linked to the increase in aggressiveness. We measured plasma corticosterone and quantified aggressive behaviour of male territorial rats at various time points of the day-night transition. Low aggression levels were observed in the full light period when plasma concentrations of corticosterone were low. An increase in plasma corticosterone occurred just prior to the dark phase, when aggressive responding was the highest. Aggressive behaviour remained high in the early dark period when corticosterone was still high. We found that blocking the high affinity mineralocorticoid receptor (MR) with spironolactone (5 or 10 mg/kg) during the early dark period dramatically and specifically reduced territorial aggression. PMID- 10792583 TI - Isolation and characterization of the promoter region of the rat vasopressin V1b receptor gene. AB - Regulation of pituitary vasopressin V1b receptors plays a critical role in regulating pituitary adrenocorticotropic hormone (ACTH) secretion during adaptation to stress. The objective of this study was to isolate the promoter regulatory region of the V1b receptor gene to better understand the molecular mechanisms involved in V1b receptor regulation. Screening of a rat genomic library using probes directed to the coding region and to the 5'UTR of the rat V1b receptor resulted in the isolation of several clones containing the 5'upstream regions of the V1b receptor cDNA. Sequencing of an 11.2 Kb fragment revealed 8.2 Kb upsteam of the reported cDNA sequence, which contains a putative promoter regulatory region. The 3' end of the clone contained 1472 base pairs corresponding to the recognized cDNA sequence, followed by 1506 bp of unknown sequence located at the end of the sixth transmembrane domain, probably corresponding to an intron, characteristic of these family of receptors. An additional 161 bp intron was found in the 5'UTR, similar to that described in the rat oxytocin receptor gene. 5'RACE and RNase protection analysis mapped two major putative transcription start points at -830 and -861 bp from the starting methionine. Analysis of the putative promoter region showed no indication of a proximal TATA box, but the presence of a CACA box, a GAGA box, several AP-1 and AP-2 sites and a cluster of Sp1 sites upstream of the AP-2 sites. A luciferase construct containing a 2.1-kb of putative promoter, and part of the 5'UTR including the first intron, showed promoter activity when transfected into COS-7, CHO and PC12 cell lines but not in AtT-20 cells. A similar construct without the intron and distal 5'UTR sequence has no promoter activity in the same cell lines. In summary, the V1b receptor gene contains at least 3 exons and 2 introns. The 5'flanking sequence contains several potential sites for transcriptional regulation, and induced luciferace activity only in constructs containing intron 1, suggesting that the latter is important for receptor gene activation. The data provide bases for future analysis of the regulatory elements controlling V1b receptor transcription. PMID- 10792585 TI - Corticosterone facilitates saccharin intake in adrenalectomized rats: does corticosterone increase stimulus salience? AB - Unlike normal rats, adrenalectomized rats do not voluntarily drink sweet saccharin solutions. To test whether this is a function of corticosterone in the circulation, and if corticosterone also increases the impetus for drinking saccharin after a period of withdrawal, we performed the following experiments. Young male rats were sham adrenalectomized (sham) or adrenalectomized (ADX); the ADX rats were provided with subcutaneous pellets containing (percent replacement of corticosterone, %B) 0%B, 15%B, 30%B or 100%B. Sham and ADX rats were immediately provided with saline (0.5%) and saccharin (2 mM) bottles in their home cages. Saccharin was allowed for 4 days on, 3 days off, 4 days on, 3 days off and a final day on, over the 15 days experiment. The dose of corticosterone determined both how much saccharin was voluntarily drunk by the ADX rats and the degree of overshoot after days off. Corticosterone also determined energy balance of the groups of ADX rats. The 30%B pellets restored food intake, body weight gain, insulin and caloric efficiency to the normal levels observed in sham rats. White fat depot weights and uncoupling protein concentration in brown adipose tissue were restored to sham levels by 100%B, suggesting that these variables which depend on activity in the sympathetic nervous system require considerable glucocorticoid receptor occupancy. We conclude that corticosterone increases the willingness to ingest sweetened water in a unimodal, dose-related manner, while moderate doses of corticosterone restore energy balance. PMID- 10792584 TI - Regional and selective effects of oestradiol and progesterone on NMDA and AMPA receptors in the rat brain. AB - We investigated the effect of 10 months ovariectomy and a correction therapy, 2 weeks before the rats were killed, of oestradiol, progesterone or their combination on NMDA and AMPA receptor binding in the hippocampus, dentate gyrus, striatum, nucleus accumbens and frontal cortex of the rat brain as well as on amino acid levels in frontal cortex. NMDA and AMPA binding densities were assayed by autoradiography using, respectively, L-[3H]glutamate and [3H]AMPA; amino acid concentrations were measured by high performance liquid chromatograhy (HPLC) coupled with UV detection. Ovariectomy was without effect on NMDA and AMPA binding density in all brain regions assayed except in the hippocampal CA1 region and dentate gyrus where it decreased NMDA binding density compared to intact rats values. Oestradiol restored and increased NMDA binding density in the CA1 subfield and the dentate gyrus of ovariectomized rats but, by contrast, it decreased binding density in the striatum and in the frontal cortex while having no effect in the CA2/3 subfield of the hippocampus and in the nucleus accumbens. Oestradiol was without effect on AMPA binding density in the hippocampus and the dentate gyrus but it reduced AMPA binding density in the striatum, the frontal cortex and the nucleus accumbens. Progesterone, and oestradiol combined with progesterone, decreased NMDA but not AMPA binding density in the frontal cortex of ovariectomized rats, and they were without effect on these receptors in the other brain regions assayed. Amino acid concentrations in the frontal cortex were unchanged after ovariectomy or steroid treatments. The effect of oestradiol in the hippocampus confirmed in the present study and our novel findings in the frontal cortex, striatum and nucleus accumbens may have functional significance for schizophrenia and neurodegenerative diseases. PMID- 10792586 TI - Voluntary sucrose ingestion, like corticosterone replacement, prevents the metabolic deficits of adrenalectomy. AB - We tested whether corticosterone replacement causes increased sucrose drinking in adrenalectomized (ADX) rats compared to sham-ADX (sham) rats. ADX rats given high doses of corticosterone drank as much sucrose as sham rats, whereas at three lower doses of corticosterone, drinking was similar between groups and was only approximately 40% of that ingested by shams. Compared to sham rats, ADX rats drinking saline, or saline and saccharin, gain weight more slowly, contain less white adipose tissue, and have higher sympathetic outflow as assessed by uncoupling protein content in brown adipose tissue. Allowing sucrose as well as saline to drink restored all of these variables to normal in ADX rats with no- or low-corticosterone. All endpoints from sucrose-drinking ADX rats with no-or low corticosterone were indistinguishable from those in water-drinking shams. By contrast, sucrose-drinking ADX rats that were given high doses of corticosterone exhibited the usual catabolic effects of corticosterone on body weight gain and, unlike sucrose-drinking shams, were obese. We conclude that (i) high corticosterone stimulates the potability of sucrose and inhibits sympathetic stimulation of uncoupling protein; (ii) sucrose, without corticosterone, normalizes metabolic deficits in ADX rats probably through actions mediated both peripherally and by the central nervous system; and (iii) ADX rats have a distinct sucrose appetite. PMID- 10792587 TI - The involvement of the styloid process in head and neck pain--a preliminary study. AB - The styloid process and associated structures have been implicated in a variety of craniomandibular dysfunctions and pain complaints. There have been anecdotal reports that treatment directed at this area can result in a dramatic reduction in referred symptoms, somatic pain and autonomic signs as well as an increase in mandibular range of motion. In the past, an elongation of the styloid process was considered necessary for pain and dysfunction symptoms to arise from this area. The patients in this study did not have elongated styloid processes, yet had orofacial pain and dysfunction symptoms seemingly referred from this area. An injection of local anaesthetic and corticosteroid in the area of the styloid process significantly reduced lateral head pain and improved mandibular function in spite of an absence of any demonstrable pathology at the styloid process. PMID- 10792589 TI - Stress distribution associated with loaded ceramic onlay restorations with different designs of marginal preparation. An FEA study. AB - The design of marginal finish of an inlay or onlay could influence the prognosis of the restoration since this area is subjected to various mechanical or chemical actions during function. This study was carried out to determine the tensile stresses which develop at the marginal area when subjected to vertical and horizontal loads. Onlay restorations with three different types of marginal finish, i. e. shoulder, chamfer and bevel, were modelled in three dimensions using the finite element method, and were then subjected to vertical and horizontal load at three different sites. The results show that horizontal forces acting on the restoration generate the highest tensile stresses whilst the vertical forces generate high tensile stresses at the margins of the chamfer and bevel designs. PMID- 10792588 TI - Comparative evaluation of secondary heat treatment and a high intensity light source for the improvement of properties of prosthetic composites. AB - This study determined the hardness and water solubility of two prosthetic composites polymerized with three curing modes for the purpose of evaluating the influence of secondary heating and a high intensity light source on resulting material properties. Two prosthetic composite materials (Artglass and Dentacolor) were cured with the following methods: (1) exposure by means of a photo-curing unit with a xenon stroboscopic light source (Dentacolor XS) for 120 s; (2) exposure with the xenon unit for 120 s, followed by heating in an oven (KL 100) at 100 degrees C for 15 min; and (3) exposure by means of a photo-curing unit with two metal halide lamps (Hyper LII) for 120 s. Knoop hardness and water solubility were determined according to standardized testing methods. Although post-cure heat treatment considerably elevated the Knoop hardness number for both materials, the greatest hardness number was generated with the use of the metal halide unit. Water solubility of the Dentacolor material was lowest when the material was cured with the metal halide unit, followed by the secondary heated group, whereas, solubility of the Artglass material was unaffected by the curing modes. This suggests that the use of a high intensity light source is more effective than the application of secondary heating for improving the post-curing properties of composites. PMID- 10792590 TI - Kinematics of the human mandible for different head postures. AB - The influence of head posture on movement paths of the incisal point (IP) and of the mandibular condyles during free open-close movements was studied. Ten persons, without craniomandibular or cervical spine disorders, participated in the study. Open close mandibular movements were recorded with the head in five postures, viz., natural head posture, forward head posture, military posture, and lateroflexion to the right and to the left side, using the Oral Kinesiologic Analysis System (OKAS-3D). This study showed that in a military head posture, the opening movement path of the incisal point is shifted anteriorly relative to the path in a natural head posture. In a forward head posture, the movement path is shifted posteriorly whereas during lateroflexion, it deviates to the side the head has moved to. Moreover, the intra-articular distance in the temporomandibular joint during closing is smaller with the head in military posture and greater in forward head posture, as compared to the natural head posture. During lateroflexion, the intra-articular distance on the ipsilateral side is smaller. The influence of head posture upon the kinematics of the mandible is probably a manifestation of differences in mandibular loading in the different head postures. PMID- 10792591 TI - Effects of four mixing methods on setting expansion and compressive strength of six commercial phosphate-bonded silica investments. AB - Six commercial phosphate-bonded silica investments were mixed with four different mixing methods such as hand, two conventional blade-driven vacuum mixers and a new planetary mixing device, and examined for their setting expansion and compressive strength. The two properties often altered, depending upon the mixing method and the investment. The setting expansion of three investments were significantly affected by the mixing method while those of the other three were not. Compressive strength of all six investments varied by changing the mixing method. It is emphasized here that a mixing method and mixing condition must be properly set for each investment to improve the fit of the final metallic castings. This study also suggests that the planetary mixing device is a useful substitute for conventional blade-driven vacuum mixers. PMID- 10792592 TI - Shear bond strength of resin teeth to heat-cured and light-cured denture base resin. AB - The failure of the bond between acrylic resin teeth and denture base material remains a considerable problem. Previous research has indicated that the introduction of a bonding agent to the tooth-resin interface significantly increased the tensile bond strength. To further investigate this finding, and to complement the earlier study, a shear strength assessment was carried out. Both a commercial and an experimental bonding agent were evaluated for tooth retention when applied to heat-cured and visible light-cured (VLC) resin. A significant increase in shear bond strength was obtained when bonding agents were applied. The experimental cement gave the greatest increase in strength, although the VLC resin failed to achieve the same degree of tooth attachment as the heat-cured resin. PMID- 10792594 TI - Effects of three concentrations of carbamide peroxide on the structure of enamel. AB - Carbamide peroxide bleaching agents have raised important questions on their potential adverse effects on the structure of enamel. The purpose of this study was to examine the effects of three carbamide peroxide bleaching agents in different concentrations (10, 16 and 35%) on the structure of enamel. Forty enamel slabs prepared from human third molars were divided equally among four groups. The specimens in the first and second group were subjected to 10% or 16% carbamide peroxide for 8 h per day for 6 weeks. Thirty-five percent carbamide peroxide was applied to the third group for 30 min a day for a 4 day period according to the manufacturers recommendation. The fourth group served as the control and was kept in artificial saliva during the test period. At the completion of the treatment, all the specimens were examined by infrared absorbtion spectroscopy and X-ray diffraction analysis. The results revealed that 10% or 16% carbamide peroxide did not seem to effect the structure of enamel, whereas 35% carbamide did affect the structure. The use of lower concentrations of carbamide peroxide (10-16%) is recommended over higher concentrations (35%) to avoid changes to the enamel. PMID- 10792593 TI - Masticatory movement in two cases with unusual alignment of the maxillary canine. AB - Masticatory function was analysed before and after orthodontic treatment in two cases where tooth alignment remained unusual after treatment. The Sirognathograph Analyzing System was used to analyse the masticatory function. In both cases, the right maxillary first premolar was located where the maxillary canine is normally positioned. The results of orthodontic treatment were satisfactory both morphologically and aesthetically, and masticatory function was greatly improved. However, some problems remained in the mandibular movement due to the abnormal contact between the mandibular teeth and the maxillary first premolar. These findings support the need for examination of masticatory function when treating patients with an unusual tooth alignment even if the results of orthodontic treatment are both morphologically and aesthetically successful. PMID- 10792595 TI - The thermal insulating effects of five dental gloves. AB - On occasions within the case reports of the various medico-legal defence organizations mention is made of burns to the oral soft tissues arising from contact with a heated instrument. Good cross infection control dictates that the dentist should be gloved whilst treating patients. No study has to date examined the thermal insulating effect of wearing dental procedure gloves although double gloving is known to blunt temperature perception. It was the purpose of this work to compare the degree of thermal insulation afforded by five makes of gloves (Biogel-D, Featherlite, Healthline, Microtouch and Tru-Touch). Measurement of temperature rises at 15, 30 and 60 s were made when a copper cylinder, at ambient room temperature, containing an iron/constantan thermocouple was placed upon a 2.35 kg aluminium block maintained at 50 degrees C by a thermostatically controlled electrical heating element. This measurement was initially performed, on 10 separate occasions, with the aluminium block and copper cylinder in direct contact (Control). This arrangement was then modified to investigate the effects of the various gloves by placing a circular mat of each glove material, harvested from the palm of each glove, between the block and cylinder. For each glove, 10 sets of observations were made using a different circular mat of glove material whose thickness had been previously determined. An analysis of variance identified highly significant (P<0.001) differences between the temperature rise of the control and experimental groups. The degree of thermal insulation afforded by each glove type appeared related to the glove thickness. This was confirmed by regression analysis but, although correlation coefficients of at least 0.91 were recorded, no single relationship best related these two quantities. Glove thermal insulating properties should be considered when selecting gloves for use in the surgery. PMID- 10792596 TI - The function of tensor veli palatini muscles in patients with aural symptoms and temporomandibular disorder. An EMG study. AB - This preliminary study investigated Myrhaug's neuromuscular hypothesis by means of audiological and electromyographic assessment. After a clinical examination of the TMJ system, the activity of the tensor veli palatini muscles in the state of rest and during swallowing was investigated in 16 patients with the help of an EMG recording, and the tube-opening function was evaluated endoscopically. Results showed that 6.25% of the patients suffered from mild joint-dysfunction (group I), 81.25% from a moderate dysfunction (group II), and 12.5% from severe dysfunction (group III). Although five patients suffered from tinnitus and 11 from fullness in the ear, both the audio- and tympanogram performed on 15 patients displayed normal recordings. Only one patient suffered from moderate sensorineural hearing loss. In the EMG recording, 11 of the 16 subjects displayed normal EMG patterns in state of rest and during swallowing contraction. In two cases, abnormal EMG recordings were obtained due to incorrect insertion of the EMG needle. The study's results indicate that spasm in the masticatory muscles of TMJ patients neither seems to cause reflex spasm of the tensor palatini muscles nor to effect significant alterations in Eustachian tube function. PMID- 10792597 TI - Resting tongue position and its relation to the state of the dentition: a pilot study. AB - The position of the resting tongue has been reportedly classified as normal or abnormal. The problems introduced by the abnormal tongue position relating to denture retention and stability have been sufficiently underlined. In the present study, the bibliography on the abnormal tongue position and its effect on denture function was reviewed. Additionally, the criteria for characterizing the tongue position as well the causes of the abnormalities, as reported in the literature, were discussed. Finally, a pilot epidemiological study was conducted, with the purpose to determine the incidence of the abnormal tongue position among edentulous patients as opposed to the dentate. The criteria used for the determination were criticized, and the mechanisms potentially leading to this habit were discussed. The findings indicate that the habit development is associated with the loss of natural teeth, therefore the mechanism relates to the functional and anatomical changes of the oral environment resulting from it. An epidemiological project has been designed, with the purpose of uncovering the associations between the observed tongue position and certain functional and anatomical features, as suggested by the present study. PMID- 10792598 TI - Regulation of human jaw tapping force with visual biofeedback. AB - The purpose of this study, which made use of visual biofeedback, was to determine whether jaw tapping force reproduction is related to the strength of tapping and to investigate how jaw tapping force affects the tapping movement curve. Nine healthy examinees were asked to reproduce jaw tapping force. We found that the ease and method of regulating jaw tapping force differed depending on the target force. We also found that jaw tapping force was regulated by alteration of the jaw opening distance, the duration of the tooth contact phase, the duration of the jaw closing phase, the maximum jaw opening velocity, and the maximum jaw closing velocity. However, the duration of the jaw opening phase and cycle time was not affected by force regulation under our experimental conditions. PMID- 10792599 TI - Corrosion resistance of a magnetic stainless steel ion-plated with titanium nitride. AB - This in vitro study evaluated the corrosion resistance of a titanium nitride (TiN) ion-plated magnetic stainless steel (447J1) for the purpose of applying a magnetic attachment system to implant-supported prostheses made of titanium. The surface hardness of the TiN ion-plated 447J1 alloy with varying TiN thickness was determined prior to the corrosion testing, and 2 micrometers thickness was confirmed to be appropriate. Ions released from the 447J1 alloy, TiN ion-plated 447J1 alloy, and titanium into a 2% lactic acid aqueous solution and 0.1 mol/L phosphate buffered saline (PBS) were determined by means of an inductively coupled plasma atomic emission spectroscopy (ICP-AES). Long-term corrosion behaviour was evaluated using a multisweep cyclic voltammetry. The ICP-AES results revealed that the 447J1 alloy released ferric ions into both media, and that the amount of released ions increased when the alloy was coupled with titanium. Although both titanium and the TiN-plated 447J1 alloy released titanium ions into lactic acid solution, ferric and chromium ions were not released from the alloy specimen for all conditions. Cyclic voltamograms indicated that the long-term corrosion resistance of the 447J1 alloy was considerably improved by ion-plating with TiN. PMID- 10792600 TI - Renoprotective benefits of RAS inhibition: from ACEI to angiotensin II antagonists. AB - In landmark clinical trials, pharmacological inhibition of the renin-angiotensin system (RAS) with angiotensin-converting enzyme inhibitors (ACEIs) attenuated the decline in renal function associated with chronic renal disease (CRD). Hemodynamic and nonhemodynamic effects of angiotensin II (Ang II) attest to its central role in the pathogenesis of CRD. Angiotensin II subtype 1 receptor antagonists (AT1RA) differ from ACEI in their effects on the RAS and on bradykinin metabolism. Elevations in bradykinin levels associated with ACEI and stimulation of angiotensin subtype 2 receptors resulting from AT1RA may produce therapeutic effects unique to each class of drug. Nevertheless, in animal models of CRD, ACEI and AT1RA exert equivalent renoprotection, implying that their renoprotective effects result primarily from inhibition of Ang II-mediated stimulation of angiotensin subtype 1 receptors. Clinical data comparing ACEI and AT1RA therapy in renal disease are limited to short-term studies, which indicate that AT1RAs have equivalent effects to ACEI on the major determinants of CRD progression, namely blood pressure and proteinuria. AT1RAs were well tolerated, with side-effect profiles similar to placebo. Taken together, available evidence suggests that AT1RAs will share the renoprotective properties of ACEI in human CRD. Nevertheless, the results of long-term clinical trials are required before AT1RA can be recommended as an alternative to ACEI in renoprotective therapy. PMID- 10792601 TI - Evidence for genetic factors in the development and progression of IgA nephropathy. AB - BACKGROUND: IgA nephropathy (IgAN) is the most common glomerulonephritis in the world among patients undergoing renal biopsy. Once considered a relatively benign condition, longitudinal follow-up studies have revealed that in fact 9 to 50% of patients progress to end-stage renal disease within 20 years of disease onset. In the three decades since its first description by Jean Berger and Nicole Hinglais, clinical, epidemiologic, and immunologic studies of the pathogenesis of primary (idiopathic) mesangial glomerulonephritis with predominant IgA deposits have characterized the features of IgAN as a distinct glomerular disease entity. However, the basic molecular mechanism(s) underlying abnormal IgA deposition in the mesangium with ensuing extracellular matrix expansion and mesangial cell proliferation remains poorly understood. The task of elucidating the molecular basis of IgAN is made especially challenging by the fact that both environmental and genetic components likely contribute to the development and progression of IgAN. METHODS AND RESULTS: We review here the evidence for genetic factors in the development and progression of IgAN, including a reappraisal of earlier conflicting results from small immunogenetic case-control studies, the evidence for racial differences in the prevalence of IgAN, a detailed summary of all reported occurrences of familial IgAN worldwide, and an exhaustive review of new insights gained through the study of two murine models of hereditary IgAN: the ddY and the uteroglobin-deficient mouse. CONCLUSIONS: With the development of powerful molecular genetic approaches to the study of both Mendelian and complex human genetic diseases, and the successful efforts of investigators to identify and clinically characterize large IgAN multiplex families, we propose that genetic analysis of familial IgAN is the most promising approach to the identification of IgAN disease/susceptibility genes. Alternatively, if the case control study design is employed to identify associations between particular candidate genes or markers and the development of IgAN, spurious associations caused by the effects of population stratification should be ruled out by confirming the findings using powerful and sensitive family-based methodologies such as the transmission/dysequilibrium test (TDT). PMID- 10792602 TI - An immunohistochemical study of developing glomeruli in human fetal kidneys. AB - BACKGROUND: In the glomerulonephritis, mesenchymal cells frequently repeat the expression of fetal immunohistochemical phenotypes. However, in human glomerulogenesis the phenotypic alteration of mesangial and other types of glomerular cells has not been clearly defined. Our aim was to clarify the characteristics of fetal mesangial cells and glomerular capillary endothelial cells, as well as their changes during glomerulogenesis using immunohistochemistry. METHODS: We examined the renal tissues of 34 autopsied fetuses and neonates, 5 children, and 5 adults using immunohistochemistry and immunoelectron microscopy, using antibodies for cytoskeletons, contraction associated proteins, and endothelial cell markers. RESULTS: In the V and S stages, there were no cells showing mesangial and endothelial features within the vesicles and the S-shaped bodies. In the S stage, small blood vessels, consisting of endothelial cells (CD31+, CD34+) and primitive perivascular mesenchymal cells (alpha-smooth muscle actin+, low molecular caldesmon+, vimentin+), were branched from developing interlobular arteries and appeared to extend to the lower clefts of the S-shaped bodies. In the C stage, the perivascular mesenchymal cells aggregated at the root of the immature glomeruli. In the M stage, they migrated toward the periphery of immature glomeruli and gradually lost their fetal immunohistochemical features. Similarly, with further maturation, the fetal glomerular capillary endothelial cells gradually lost the immunostaining for CD34, while the strong staining intensity of CD31 remained unchanged, just as that in the adult glomerular capillary endothelial cells. CONCLUSIONS: In human glomerulogenesis, we demonstrate that fetal mesangial and capillary endothelial cells change their immunohistochemical phenotypes with maturation. They gradually lose fetal immunohistochemical phenotypes. Already before birth, the mesangial cells in almost all glomeruli at the late M stage acquire the adult phenotype. PMID- 10792603 TI - Molecular cloning, expression, and distribution of glomerular epithelial protein 1 in developing mouse kidney. AB - BACKGROUND: Glomerular epithelial protein 1 (GLEPP1) is a receptor-like membrane protein tyrosine phosphatase (RPTP) with a large ectodomain consisting of multiple fibronectin type III repeats, a single transmembrane segment, and a single cytoplasmic phosphatase active site sequence. In adult human and rabbit kidneys, GLEPP1 is found exclusively on apical membranes of podocytes and especially on surfaces of foot processes. Although neither ligand nor function for this protein is known, other RPTPs with similar topologies have been implicated in mediating adherence behavior of cells. METHODS: To evaluate potential roles of GLEPP1 further, we cloned the full-length mouse GLEPP1 cDNA and examined its expression patterns in developing kidney by Northern blot analysis, in situ hybridization, and immunofluorescence microscopy. RESULTS: Nucleotide sequencing showed that mouse GLEPP1 was approximately 80% identical to rabbit and human GLEPP1 and approximately 91% identical at the amino acid level. The membrane-spanning and phosphatase domains of mouse GLEPP1 shared> 99% homology with PTPphi, a murine macrophage cytoplasmic phosphatase. Northern analysis identified a single GLEPP1 transcript of approximately 5.5 kb in fetal kidney that became approximately threefold more abundant in adults. In situ hybridization of newborn mouse kidney revealed GLEPP1 mRNA in visceral epithelial cells (developing podocytes) of comma- and S-shaped nephric figures, and expression increased in capillary loop and maturing stage glomeruli. Beginning on embryonic day 14, GLEPP1 protein was first observed on cuboidal podocytes of capillary loop stage glomeruli, but nascent podocytes of earlier comma- and S shaped nephric figures were negative. At later stages of glomerular maturation, where foot process elongation and interdigitation occurs, GLEPP1 immunolabeling intensified on podocytes and then persisted at high levels in fully developed glomeruli. CONCLUSION: Our findings are consistent with a role for GLEPP1 in mediating and maintaining podocyte differentiation specifically. PMID- 10792604 TI - Role of fibronectin deposition in branching morphogenesis of Madin-Darby canine kidney cells. AB - BACKGROUND: Madin-Darby canine kidney (MDCK) epithelial cells grown in collagen gels in the presence of hepatocyte growth factor (HGF) form branching tubules. The tubule-lining epithelial cells are polarized with the basolateral surface in contact with the collagen gel and the apical surface facing the lumen. To delineate whether MDCK branching tubules construct the basal lamina, we characterized the composition of the extracellular matrix deposited by MDCK tubules. The tubule-lining cells produced an apparently incomplete basal lamina containing a discontinuous laminin substratum. In addition, a thick layer of fibronectin surrounded the basal cell surface of the branching tubule. In an attempt to delineate the role of fibronectin deposition in branching morphogenesis, we conducted this study. METHODS: MDCK cells cultured in collagen gel were employed. We first used arginine-glycine-aspartate peptides containing disintegrin rhodostomin to disturb the interactions between fibronectin and cell surface integrins. Furthermore, we established several stable transfectants expressing fibronectin antisense RNA to examine the role of fibronectin in branching morphogenesis directly. RESULTS: Rhodostomin inhibited the formation of branching tubules. The transfectants expressing fibronectin antisense RNA exhibited relatively lower levels of synthesized fibronectin and markedly diminished growth rates of branching tubules than the control clone. An inhibition of branching morphogenesis induced by the overexpression of fibronectin antisense RNA was manifested by the decrease in cell growth rates and cell migration. CONCLUSION: These results indicate that the deposition of fibronectin underlying the tubule-lining epithelium serves to enhance cell proliferation and migration, and hence facilitates the branching tubulogenesis of MDCK cells. PMID- 10792605 TI - WT1 splice-site mutations are rarely associated with primary steroid-resistant focal and segmental glomerulosclerosis. AB - BACKGROUND: Donor splice-site de novo heterozygous mutations in intron 9 of the Wilms' tumor gene (WT1) have been reported in Frasier syndrome, which is defined by the association of focal and segmental glomerulosclerosis (FSGS), male pseudohermaphroditism, and gonadoblastoma. These splice-site mutations alter the WT1 alternative splicing leading to two WT1 isoforms, with (+) or without (-) three amino acids, lysine-threonine-serine (KTS), between zinc fingers 3 and 4. The aim of this work was to investigate the possibility that some cases of primary steroid-resistant nephrotic syndrome associated with FSGS may be caused by WT1 splice-site mutations. METHODS: We analyzed WT1 exons 8 and 9 and the surrounding exon/intron boundary DNA sequences in 37 children with nonfamilial primary steroid-resistant nephrotic syndrome. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the relative ratio of +KTS/-KTS transcripts from immortalized lymphocyte RNA. RESULTS: One boy with FSGS and associated pathologies (diaphragmatic hernia, proximal hypospadias, and unilateral testicular ectopia) was found to carry the heterozygous 1228 +4 C-->T splice-site mutation. RT-PCR quantitation of the +KTS/ KTS transcripts from immortalized lymphocyte RNA of this patient showed a diminution of the +KTS/-KTS isoform ratio (0.43), which is identical to that reported in patients with Frasier syndrome. Using the same approach, healthy control subjects have +KTS/-KTS ratios ranging from 1.50 to 2.00. CONCLUSIONS: This study expands the range of the phenotypic presentation of the intron 9 splice-site WT1 mutations and adds to the already reported heterogeneity of primary steroid-resistant nephrotic syndromes. We suggest that these mutations are not likely to be a common cause of isolated steroid-resistant nephrotic syndrome, and recommend a WT1 exon 9/intron 9 splice-site study in children with primary steroid-resistant nephrotic syndrome if genital or diaphragmatic anomalies are associated. The identification of such WT1 mutations has practical implications for the management of these patients. PMID- 10792606 TI - Expression of p300-truncated fragments results in the modulation of apoptosis in rat mesangial cells. AB - BACKGROUND: Mesangial cell proliferation, apoptosis, and matrix deposition have pivotal roles in the pathogenesis of renal diseases such as diabetic nephropathy and glomerulonephritis. The behavior of mesangial cells depends on the integration of intracellular signals elicited by hormones and cytokines. We hypothesized that p300 is primarily involved in the integration of signal transduction pathways in rat mesangial cells (RMCs) and that interference with p300 function will alter apoptotic signals. METHODS: We established an RMC cell line expressing the Tet-activator (tTA). RMC-tTA cells were transiently transfected with vectors coding for either the N-terminal third or the C-terminal third of p300. Expression was induced by the addition of doxycycline [Dox; 1 microg/mL; 5% fetal bovine serum (FBS)]. The percentage of apoptosis was determined using the TUNEL technique. Specific protein-protein interactions were determined by Western blot analysis of immunoprecipitated complexes. Cells were treated with 5% FBS or with H2O2 (500 micromol/L, 1 h) with and without Dox. RESULTS: The expression of p300-C resulted in increased susceptibility to low serum-induced (20.0 +/- 4.6 vs. 3.0 +/- 1.7%) and to H2O2-induced apoptosis (75.3 +/- 13.3 vs. 50.8 +/- 6.5%) compared with controls. Immunoprecipitation of p300-C showed an interaction with the transcription factor c-Fos, which was enhanced by H2O2 treatment. Expression of the p300-N resulted in a rescue (34.8 +/- 6. 4 vs. 50.8 +/- 6.5%) from H2O2-induced apoptosis compared with controls. P300-N was shown to form a complex with the transcription factor nuclear factor-kappaB (NF kappaB). CONCLUSIONS: The data indicate that endogenous p300 is involved in apoptosis in mesangial cells. We propose that interference or enhancement of endogenous p300 function, by expression of exogenous fragments, can alter interactions with c-Fos or NF-kappaB and modulate signals during cellular stress. PMID- 10792607 TI - Effects of endothelin or angiotensin II receptor blockade on diabetes in the transgenic (mRen-2)27 rat. AB - BACKGROUND: Endothelin (ET) and angiotensin II (Ang II) are vasoactive/trophic peptides that may contribute to the progression of diabetic nephropathy. The transgenic (mRen-2)27 rat exhibits overexpression of Ang II at sites of normal physiological expression. Unlike other rat strains, the streptozotocin-induced diabetic Ren-2 rat develops progressive renal pathology associated with a declining glomerular filtration rate (GFR) and provides a convenient model to evaluate the role of these vasoactive peptides in the nephropathic process. METHODS AND RESULTS: Oral administration of either the endothelin A (ETA) and ETB receptor antagonist bosentan or the angiotensin type 1 (AT1) receptor antagonist valsartan for 12 weeks reduced systolic blood pressure (SBP) of nondiabetic and diabetic Ren-2 rats to normotensive levels. Diabetic renal pathology was associated with intense renin mRNA and protein in the proximal tubules and juxtaglomerular cells along with overexpression of transforming growth factor beta1 (TGF-beta1) and collagen IV mRNA in glomeruli and tubules. With valsartan but not bosentan, renin mRNA and protein in the proximal tubules were not detected. Valsartan but not bosentan reduced TGF-beta1 and collagen IV mRNA and the severity of diabetic renal pathology. A declining GFR with diabetes was attenuated by both treatments. Albuminuria in diabetic rats rose further with bosentan but was reduced with valsartan. CONCLUSIONS: Despite producing normotension, severe diabetic renal pathology was not prevented by bosentan, suggesting dissociation of ET, albuminuria, and hypertension from the structural injury in this diabetic model. The beneficial effects afforded by valsartan therapy strengthen the importance of the local renin-angiotensin system in mediating progressive diabetic renal injury. PMID- 10792608 TI - Exposure of endothelial cells to recombinant human erythropoietin induces nitric oxide synthase activity. AB - BACKGROUND: Anemic patients with chronic renal failure receiving recombinant human erythropoietin (rHuEPO) therapy frequently develop hypertension through an unknown mechanism. We hypothesize that EPO receptors (EPORs) on endothelial cells (ECs) in various sites of vasculature may mediate the activities of nitric oxide synthase (NOS) and/or the release of endothelin-1 (ET-1), contributing to blood pressure changes. We tested this hypothesis using primary cultures of ECs obtained from human coronary artery (HCAEC), pulmonary artery (HPAEC), dermis (HDEC), and umbilical vein (HUVEC). METHODS: EPORs were measured by 125I-EPO binding. The effect of EPO on EPOR, ET-1, and NOS mRNA levels was assessed by quantitative reverse transcription-polymerase chain reaction. Cellular NOS activity and ET-1 release into the medium was measured by the NOSdetect assay and by radioimmunoassay kits. RESULTS: Short-term (4 h) treatment with EPO (4 U/mL) did not change the number or affinity of EPOR per cell. Neither were there any changes in the amount of EPOR, ET-1, and NOS transcripts (cDNA/microg of mRNA) nor in ET-1 release and NOS activity. In HUVEC only, 24-hour exposure to EPO caused a threefold increase in NOS transcript. In other cells, EPO treatment for six days increased NOS activity by twofold to fourfold. CONCLUSIONS: We show that upon extended exposure, EPO induces NOS activity but does not affect ET-1 release. These findings indicate that the hypertensive effect of EPO is not likely to be caused by a direct effect on ECs. PMID- 10792609 TI - Abnormal platelet cytoskeletal assembly in hemodialyzed patients results in deficient tyrosine phosphorylation signaling. AB - BACKGROUND: Uremic patients have a bleeding tendency associated with a platelet dysfunction. We evaluated the impact of a repeated hemodialysis procedure on primary hemostasis by analyzing different aspects of platelet activation in uremic patients. METHODS: Studies were performed in (1) eight patients with end stage renal disease before the hemodialysis program was initiated and after initiating hemodialysis treatment, and in (2) eight patients on maintenance hemodialysis who were transferred to continuous ambulatory peritoneal dialysis. Studies included routine platelet aggregations and evaluation of platelet subendothelium interactions under flow conditions. Contractile proteins and tyrosine phosphorylation associated with the cytoskeleton were analyzed, before and after thrombin activation of platelets, by electrophoresis after Triton X-100 extraction. RESULTS: No changes in the clinical parameters analyzed were observed among the different study groups. Aggregation and platelet adhesion only improved when patients were shifted from hemodialysis to continuous ambulatory peritoneal dialysis (P < 0.05 for both percentage of surface covered by platelets and aggregate formation). The association of cytoskeletal proteins in platelets from patients under hemodialysis treatment was statistically decreased with respect to the corresponding values in platelets from patients not subjected to dialysis (P < 0.01 for actin). However, after two months on peritoneal dialysis, these values increased to almost control values (P < 0.001 for actin, vs. hemodialysis). Similarly, translocation of tyrosine-phosphorylated proteins to the cytoskeletal fraction was impaired in platelets from hemodialyzed patients, and it recovered partially after the patients transferred to continuous ambulatory peritoneal dialysis. CONCLUSIONS: Our present data support the concept that repeated platelet stress during hemodialysis has a deleterious effect on the organization of platelet cytoskeleton, which seems to impair the translocation of signal transduction proteins within platelets compromising the platelet function in uremia. PMID- 10792610 TI - PDGF regulates gap junction communication and connexin43 phosphorylation by PI 3 kinase in mesangial cells. AB - BACKGROUND: Gap junctional intercellular communication (GJIC) plays an important role in the regulation of cell growth, migration, and differentiation. Ultrastructural and histochemical studies indicate the existence of a high density of gap junctions among mesangial cells (MCs), but little is known about their regulation. Because of the close link between growth and GJIC, we examined how platelet-derived growth factor (PDGF) may affect GJIC in cultured MCs. METHODS: MCs were exposed to PDGF in the presence or absence of phosphatidylinositol 3' kinase (PI3K) inhibitors, and GJIC was evaluated by the transfer of Lucifer yellow. The gap junction protein connexin43 (Cx43) was examined by immunohistochemistry, immunoprecipitation, and Western blot. RESULTS: The addition of PDGF into MC culture caused a rapid and transient inhibition of GJIC, with maximal inhibition (80%) occurring 15 minutes after PDGF exposure and returning to control levels after 90 minutes. This action of PDGF could be largely prevented by pretreatment of MCs with the PI3K inhibitor LY294002. Immunochemical staining showed that PDGF did not alter the localization and distribution of Cx43. Immunoprecipitation studies demonstrated that PDGF induced a rapid and transient increase of tyrosine phosphorylation of Cx43 protein, which was dose dependent and in accordance with the time course of the disruption of GJIC. PDGF also elicited activation of extracellular signal-regulated kinase (ERK). Using two structurally unrelated PI3K inhibitors, wortmanin and LY294002, both tyrosine phosphorylation of Cx43 and activation of ERK stimulated by PDGF were largely blocked. CONCLUSION: These results suggest that PDGF abrogates GJIC function in MCs via the PI3K-dependent signaling pathway. Disruption of GJIC by PDGF could be one mechanism by which PDGF modulates MC behavior. Participation of PI3K in the regulation of GJIC demonstrates the complex coordination of molecular events that accompany MC mitogenesis. PMID- 10792611 TI - Estrogen worsens incipient hypertriglyceridemic glomerular injury in the obese Zucker rat. AB - BACKGROUND: The obese Zucker rat (OZR) is a model of glomerulosclerosis and renal failure in the setting of hyperlipidemia, hyperinsulinemia, and obesity. Our prior work in OZRs has shown that ovariectomy attenuates glomerulosclerosis, while added estrogen worsens it. To investigate the mechanism of estrogen's effects on glomerular disease in this model, we evaluated the effects of ovariectomy and estrogen supplementation on seven-week peripubertal OZRs. At this time point, rats exhibit no overt histologic glomerular disease, but are just beginning to show elevated urinary albumin excretion. METHODS: Female OZRs fed ad libitum were ovariectomized at four weeks, with or without estrogen supplementation to raise estrogen levels to just below those of preoestral adults (mean 16.5 pg/mL). Sham-operated controls were included. RESULTS: Ovariectomy normalized albuminuria, lowered total and very low-density lipoprotein triglycerides, and reduced glomerular fibronectin expression. Estrogen supplementation worsened albuminuria and raised total/very low-density lipoprotein triglycerides and total cholesterol. Estrogen-supplemented rats exhibited enhanced glomerular deposition of apo A-IV and apo B, increased glomerular expression of desmin and type IV collagen, and increased interstitial macrophage deposition. CONCLUSION: Estrogen may be permissive for the early development of renal disease in OZRs and may act by increasing triglyceride-rich lipoproteins, which then bind to glomerular cells and initiate or accelerate glomerulosclerosis. PMID- 10792612 TI - Identification of a novel Fcalpha receptor expressed by human mesangial cells. AB - BACKGROUND: IgA nephropathy (IgAN) is characterized by mesangial deposits of polymeric IgA (pIgA). The pathological consequences of IgA deposition are believed to center on direct interaction between IgA and the glomerular mesangial cell (MC). We have characterized a novel mesangial receptor that recognizes the Fc portion of IgA. METHODS: Five primary MC cultures were evaluated for IgA binding by flow cytometry, and specificity of binding was determined by competitive inhibition. Relative affinities of the receptor for all IgA isoforms were also determined, and binding of pIgA1 was compared to monomer. The identified Fc receptor was then compared with CD89, hitherto the only other Fcalpha receptor reported. CD89 protein and mRNA expression were detected by conventional and intracellular flow cytometry, sequencing of reverse transcription-polymerase chain reaction (RT-PCR) products, and Northern blotting. RESULTS: All MCs constitutively expressed a receptor that bound IgA in an Fcalpha dependent fashion. The receptor recognized secretory and serum IgA1 and IgA2 equally, but pIgA bound with much greater affinity than monomer. At no time were we able to detect CD89 synthesis, although three novel CD89-related mRNA transcripts were identified by RT-PCR. CONCLUSIONS: We have clearly demonstrated that MCs consistently express an FcalphaR distinct from the myeloid FcalphaR CD89. This novel receptor binds pIgA with high affinity and may therefore mediate the mesangial injury that follows IgA deposition in IgAN. While immunogenically distinct, the mesangial Fcalpha receptor may share some molecular homology with CD89, as mRNA transcripts with partial identity to CD89 were found in all five MC cultures. PMID- 10792613 TI - Cloning of rat nephrin: expression in developing glomeruli and in proteinuric states. AB - BACKGROUND: Nephrin is identified as a product of the gene mutated in a patient with congenital nephrotic syndrome of the Finnish type. However, its precise localization and function are not yet fully clarified. METHODS: To clone the rat homologue of nephrin, polymerase chain reaction (PCR) was employed. To elucidate the localization and expression of nephrin, immunohistological analysis with a specific antirat nephrin antibody, reverse transcription-PCR, and RNase protection assay were performed. RESULTS: Amino acid sequences of rat and human nephrin are highly homologous (82.2% identity). The domain structure of nephrin is also highly conserved between rats and humans. The rat nephrin was detected only in kidney glomeruli along glomerular capillary walls, and its localization was always identical to that of the anti-slit diaphragm monoclonal antibody (mAb) 5-1-6-recognized antigen in normal matured and fetal rat glomeruli and in the glomeruli of proteinuric states. The nephrin staining pattern was clearly distinguished from that of zonula occludens-1 (ZO-1), alpha3-integrin, or podocalyxin. mRNA expression for nephrin was first detected in the fetal rat kidneys at 18.5 embryonic days. Nephrin mRNA expression decreased just after injection of mAb 5-1-6 (47.4%) or puromycin aminonucleoside (51.2%), and the staining pattern of nephrin shifted from a linear to a granular pattern in both proteinuric states. CONCLUSIONS: Nephrin is localized in slit diaphragm in the matured glomeruli and is identical with mAb 5-1-6 antigen. Nephrin is involved in the development of proteinuria not only in mAb 5-1-6 nephropathy, but also in puromycin aminonucleoside nephropathy. PMID- 10792614 TI - Proximal tubular epithelial hyperplasia in patients with chronic glomerular proteinuria. AB - BACKGROUND: Proteinuria is known to affect the proximal tubular epithelial structure and function. The present study tested the hypothesis that chronic proteinuria leads to hyperplasia of proximal tubular epithelium. METHODS: This hypothesis was tested by morphometric analysis of the renal biopsy specimens in two groups of patients. Group A (N = 15) was composed of patients with chronic glomerular proteinuria who, for clinical indications, underwent renal biopsy of their native kidneys on two separate occasions. The proteinuria was sustained during the first and second renal biopsies in all but two of the patients with minimal change nephrotic syndrome who experienced transient remission. Group B (N = 10) was composed of patients with little or no proteinuria who underwent renal biopsy because of unexplained hematuria and whose renal biopsy showed only thin glomerular basement membrane (GBM) disease. RESULTS: In Group A, the mean number of epithelial cell nuclei per proximal tubule cross-section increased significantly from the first to the second renal biopsy (11.0 +/- 2.7 vs. 13.0 +/ 2.2, P = 0.005, paired t-test). Also, those with severe proteinuria showed proximal tubules with reactive epithelium (large pale nuclei with a high nucleus to cytoplasm ratio) and marked hyperplasia (double and triple layers of epithelium). Such changes were not seen in group B renal biopsies. Compared with group A biopsies, group B biopsies showed a lower mean value for proximal tubular epithelial cell nuclei per tubular cross-section (P = 0.056) and a higher mean proximal tubular volume (P = 0.049). As a consequence, the mean number of nuclei per relative tubular volume was significantly greater in group A compared with group B (0.55 +/- 0.14 vs. 0.40 +/- 0.06, P = 0.003, by Wilcoxon rank sum). CONCLUSIONS: Chronic heavy proteinuria is associated with hyperplasia of proximal tubular epithelium and contraction of proximal tubular volume. These events may impair glomerular filtration and represent another mechanism of progression of renal disease. PMID- 10792615 TI - Increased nitrotyrosine staining in kidneys from patients with diabetic nephropathy. AB - BACKGROUND: Proximal tubular cells produce nitric oxide (NO.). We have shown that under hyperglycemic conditions, cultured proximal tubular cells express cytochrome P450 2E1, which is capable of producing superoxide (O2.). NO. and O2. react to form peroxynitrite (ONOO.), a powerful oxidant. ONOO. nitrosylates tyrosine moieties on proteins causing tissue damage. Our hypothesis is that ONOO. plays a role in early diabetic tubular damage and perhaps disease progression. METHODS: Renal biopsies from patients with diabetic nephropathy (DM), acute allograft rejection (AAR), acute allograft tubular necrosis (ATN), and glomerulonephritis (GN) were obtained. Normal kidney specimens were taken from nephrectomy samples (N = 10 for each group). The tissues were examined for the presence of nitrotyrosine using an immunoperoxidase technique with a polyclonal antibody. Samples were then arbitrarily scored, and the results analyzed (analysis of variance and Student's t-test for unpaired data). The number of apoptotic cells in a sample of tubules in each biopsy was also assessed. RESULTS: The DM biopsies showed increased staining for nitrotyrosine in proximal tubules (P = 0.0001) and in the thin limb of the loop of Henle (P = 0.0006) compared with all other groups. There was increased staining in the ascending and distal tubules in GN as compared to DM and ATN (P = 0.01). Nitrotyrosine was also found in all distal tubules and collecting ducts, including normals. There was no difference in the number of apoptotic tubular cells in diabetics compared with controls. CONCLUSION: To our knowledge, these data provide the first evidence for the presence of nitrotyrosine in both normal and diseased kidneys. The significance of the findings in normals is unclear, but could be due to activation of constitutive NOS. However, the study clearly demonstrates increased production of ONOO. in proximal tubules of patients with DM, and suggests that oxidant injury of the proximal tubules plays an important part in the pathogenesis of DM. PMID- 10792616 TI - Gene transfer targeting interstitial fibroblasts by the artificial viral envelope type hemagglutinating virus of Japan liposome method. AB - BACKGROUND: Tubulointerstitial inflammation and fibrosis are commonly associated with most human glomerular diseases. The degree of tubulointerstitial damage, rather than the glomerular injury, could correlate with the degree of renal functional impairment and accurately predict long-term prognosis. In an effort to understand the pathogenesis of the progressive interstitial fibrosis, we developed a new strategy of gene transfer to the interstitial fibroblasts. METHODS: Either fluorescein isothiocyanate (FITC)-labeled oligodeoxynucleotides (ODNs) or pEBAct-NlacF expression vector was introduced into the kidney of normal rats retrogradely via ureter by using the artificial viral envelope (AVE)-type hemagglutinating virus of Japan (HVJ) liposome method. RESULTS: FITC-labeled ODNs were accumulated diffusely in the nuclei of the interstitial cells in the transfected kidney 10 minutes after transfection, and the interstitial cells were identified as interstitial fibroblasts by immunostaining with ER-TR7. To examine the gene expression in the interstitium, pEBAct-NlacF gene-conjugated HVJ liposome was injected retrogradely through the ureter, and in consequence, nuclear beta-galactosidase activity was continuously observed in interstitial cells at least two weeks after transfection. CONCLUSION: This new strategy of gene transfer to the interstitial fibroblasts is useful for the investigation of the pathophysiology of tubulointerstitial lesion, and furthermore, it may be a promising new therapeutic method for the progression of interstitial fibrosis. PMID- 10792617 TI - ANCA antigens, proteinase 3 and myeloperoxidase, are not expressed in endothelial cells. AB - BACKGROUND: One hypothesis for the pathogenesis of vasculitis associated with antineutrophil cytoplasmic autoantibodies (ANCAs) proposes that ANCAs bind to ANCA antigens, such as proteinase 3 (PR3) or myeloperoxidase (MPO), which are produced by endothelial cells and expressed on their surfaces. There are conflicting reports, however, on whether endothelial cells express the ANCA antigen PR3, and there are no reports on endothelial expression of MPO. The aim of this study was to determine the presence or absence of PR3 and MPO mRNA in both venous and arterial endothelial cells, employing standard reverse transcription-polymerase chain reaction (RT-PCR) techniques and also the quantitative and highly specific method, TaqMan PCR. METHODS: RT-PCR (with 3 primer sets) and TaqMan PCR, a method for detecting low copy transcripts, were used to probe for PR3 and MPO transcripts in human endothelial cells from umbilical vein (HUVEC) and artery (HUAEC) and from lung microvascular (HLMVEC). Cells were treated with interferon-gamma (200 units/mL) or tumor necrosis factor alpha (3 or 10 ng/mL) or both. RESULTS: Transcripts for PR3 and/or MPO were not detected in HUVEC, HUAEC, and HLMVEC by standard RT-PCR. Analyses for PR3 protein confirmed that PR3 is not expressed in HUVEC. HUVEC and HUAEC were negative for PR3 and MPO by TaqMan PCR. CONCLUSIONS: PR3 and MPO are not expressed in HUVEC, HUAEC, or HLMVEC. Endothelial cell presentation of endogenous PR3 and MPO antigens is not involved in the pathogenesis of ANCA-associated vasculitis. Alternative explanations need to be explored to determine the pathogenic effect of ANCAs. PMID- 10792618 TI - IGFBP-5(201-218) stimulates Cdc42GAP aggregation and filopodia formationin migrating mesangial cells. AB - BACKGROUND: We have previously shown that insulin-like growth factor-I (IGF-I) and IGF binding protein-5 (IGFBP-5) induce mesangial cell migration using separate stimulatory and effector pathways. The IGFBP-5 stimulatory pathway is mediated by the serine/threonine kinase IGFBP-5 receptor, which is activated by the carboxy-terminal peptide IGFBP-5201-218. In this study, we examined the direct effects of IGFBP-5201-218 on stimulatory and effector pathways that lead to a change in mesangial cell (MC) phenotype. METHODS: Rapid actin reorganization, formation of filopodia, and characterization of novel substratum attachment structures that develop during IGFBP-5-mediating migration were examined by light, immunofluorescence, and electron microscopy. Using a wounding assay, migration was measured after the addition of stimulants and inhibitors. RESULTS: Stimulation of MCs with IGFBP-5201-218 induces rapid actin reorganization and loss of peripheral focal adhesions. The MCs develop long cellular extensions where f-actin and beta-actin terminate in unique substratum attachments. Fluorescence microscopy of stimulated cells shows that Cdc42GAP aggregates within minutes following treatment with IGFBP-5201-218. In contrast, IGF-I increases staining for Rac-1, but not Cdc42GAP, in association with the formation of prominent leading lamellae without filopodia. Staurosporin inhibits cell migration and Cdc42GAP aggregation only when added within the first hour, suggesting that it inhibits the stimulatory effect of IGFBP-5201-218 by blocking the IGFBP-5 receptor serine/threonine kinase activity. CONCLUSIONS: These data demonstrate that IGFBP-5201-218 preferentially activates Cdc42 and induces the formation of long filopodia with unique substratum attachments that produce a novel mode of locomotion. PMID- 10792619 TI - Expression of lactoferrin in the kidney: implications for innate immunity and iron metabolism. AB - BACKGROUND: Sequestering of free iron by lactoferrin (LF) is important in the defense against bacteria. In a screening for LF expression in various organs, high levels of LF mRNA were detected in human kidney. This indicated that LF is produced by the kidney and that it may participate in innate immunity of this organ. METHODS AND RESULTS: Antibody staining and in situ hybridization of paraffin-embedded kidney sections showed that LF is expressed in cells lining the distal collecting ducts of the medulla. High levels of both protein and mRNA were detected in these cells. However, a clear difference in the distribution of mRNA and protein within the tissue was observed. LF mRNA was detected along a relatively large portion of the tubuli, whereas LF antigen was found mainly in the very distal regions of the same tubuli. This indicates that LF is released by large regions of the tubuli and possibly reabsorbed in the most distal parts. Using enzyme-linked immunosorbent assay, only very low LF levels were detected in urine. CONCLUSION: The present study shows that LF is produced by the kidney and that both LF mRNA and protein are distributed in a highly ordered fashion. This latter finding, together with the very low levels of LF detected in urine, indicates that LF may contribute to the immune defense in the kidney by reduction of available free iron in the urine. Other possibilities are that LF may play a role in the iron metabolism by recovering free iron from urine and making it available for metabolic use, and that LF may participate in the antioxidant defense systems protecting the kidney against nonmicrobial oxidative injury, that is, ischemia, reperfusion and inflammation. PMID- 10792620 TI - Lipopolysaccharide-induced MCP-1 gene expression in rat tubular epithelial cells is nuclear factor-kappaB dependent. AB - BACKGROUND: Endotoxin is an important factor in the development of acute renal failure related to infection and in acceleration of chronic nephritis. Lipopolysaccharide (LPS; the major component of endotoxin) is one of the most potent triggers for renal cells to produce monocyte chemoattractant protein-1 (MCP-1), a key cytokine involved in immune cell recruitment into the renal interstitium in acute and chronic renal diseases. Knowledge about the transcriptional regulation of MCP-1 in renal tubular epithelial cells in response to LPS is incomplete. METHODS: Transcriptional regulation of MCP-1 was investigated in rat proximal tubule cells (PTCs) in primary culture and was exposed to LPS using electromobility shift assay and supershift analysis for nuclear factor-kappaB (NF-kappaB) and Western blot for the NF-kappaB inhibitory protein IkappaB. To prove the role for NF-kappaB, activator protein (AP-1), and sequence-specific transcription factor (Sp1), mutation and deletion analysis was performed using a 3.5 kb fragment of rat MCP-1 5'-flanking region inserted into a luciferase reporter construct transfected into tubular epithelial cell line (NRK 52E). RESULTS: LPS increased NF-kappaB in a dose- and time-dependent manner, which paralleled that of MCP-1 mRNA expression. IkappaBalpha decreased within 30 minutes of LPS treatment, but returned to basal levels by two hours. IkappaBbeta levels were depressed within one hour and remained low throughout the culture period after LPS stimulation. The activity of the transfected 5'-flanking region of the MCP-1 gene increased nearly threefold after LPS stimulation. Mutation or deletion of NF-kappaB binding sites, located in the enhancer region of the 5' flanking region, resulted in a total loss of LPS-induced increase in luciferase activity. Mutation of putative AP-1 and Sp1 sites, located in the proximal promoter region of MCP-1, reduced basal luciferase activity in unstimulated cells, but had no effect on LPS-stimulated luciferase activity. CONCLUSIONS: These studies prove that NF-kappaB is critical for LPS-induced MCP-1 transcription, and AP-1 and Sp1 are essential for basal expression of MCP-1 in rat tubule cells. The species-specific nature of transcriptional regulation of MCP-1 has important implications for the delineation of treatment to prevent endotoxin-mediated renal injury. PMID- 10792621 TI - beta2-microglobulin induces MMP-1 but not TIMP-1 expression in human synovial fibroblasts. AB - BACKGROUND: beta2-Microglobulin (beta2m) amyloidosis is a destructive articular disease that causes significant morbidity in patients undergoing hemodialysis. The amyloid deposits contain beta2m, some of which is altered with advanced glycation end products (AGE-beta2m). The deposits are located principally in joint structures, with adjacent degradation of cartilage and bone. We hypothesized that one of the mechanisms by which beta2m induces joint destruction is to induce the release of matrix metalloproteinase-1 (MMP-1), but not tissue inhibitor of metalloproteinase-1 (TIMP-1), from synovial fibroblasts. METHODS: To test this hypothesis and determine the role of AGE-beta2m, we incubated human osteoarthritic synovial fibroblasts in the presence and absence of beta2m and AGE beta2m and measured the release of interstitial collagenase (MMP-1) and/or TIMP-1 by enzyme-linked immunosorbent assay and Northern blot analysis. RESULTS: beta2m and AGE-beta2m at 10 and 25 microg/mL induced the release of MMP-1 from human osteoarthritic synovial fibroblasts at 24 hours. In contrast, there was no increased release of TIMP-1, leading to an increase in the MMP-1/TIMP-1 ratio indicative of uncontrolled collagenolysis. A similar dose response was observed at 48 hours, except that AGE-beta2m had no effect over control cultures. MMP-1 mRNA expression by Northern blot analysis paralleled these findings. The source of the fibroblasts did not alter the results. Finally, we demonstrated that doxycycline, a treatment for arthritis, can inhibit the release of MMP-1 from synovial fibroblasts incubated with beta2m. CONCLUSION: beta2m, at physiologically relevant concentrations, induces the release of MMP-1 without concomitant release of TIMP-1 from human synovial fibroblasts, leading to uncontrolled collagenolysis. The alteration of beta2m with AGE did not alter this effect at 24 hours, but blocked the effect at 48 hours. These findings may account for the tissue destruction seen in beta2m amyloidosis. PMID- 10792622 TI - Regulation of mouse podocyte process dynamics by protein tyrosine phosphatases rapid communication. AB - BACKGROUND: Effacement of podocyte foot processes occurs early in many glomerular diseases associated with proteinuria and is accompanied by a reorganization of the actin cytoskeleton. The molecular mechanisms regulating these structural changes are poorly understood. METHODS: To address these questions, we analyzed the effect of the polycation, protamine sulfate (PS), and puromycin aminonucleoside (PA) on the morphology, cytoskeleton, and tyrosine phosphorylation of differentiated process-bearing cultured podocytes. RESULTS: PS and PA induced similar profound morphological alterations, including retraction and detachment of podocyte processes from the extracellular matrix (ECM). The effects of PS occurred within six hours, whereas PA showed its most severe effects after 72 hours. Structural changes included reorganization of the actin cytoskeleton and focal contacts and were accompanied by an increase in tyrosine phosphorylation. The same effects were induced by application of vanadate, an inhibitor of protein tyrosine phosphatases (PTPs), suggesting that PTPs regulate podocyte process structure. Since disruption of the actin cytoskeleton with cytochalasin B protected the cells from PS-induced effacement and detachment, cytoplasmic PTPs were implicated in these events. Using reverse transcription polymerase chain reaction (RT-PCR), we demonstrated the expression of four cytoplasmic PTPs in podocytes: SHP-2, PTP-PEST, PTP-1B, and PTP-36. CONCLUSIONS: These studies indicate an important role for cytoplasmic PTPs as regulators of podocyte process dynamics. Future studies will aim at restoring the normal foot process architecture of podocytes in glomerular diseases associated with proteinuria by modulating the activity of cytoplasmic PTPs. PMID- 10792623 TI - Transport of thiamin in rat renal brush border membrane vesicles. AB - BACKGROUND: Unlike intestinal absorption, renal transport of thiamin has received little attention. This study was designed to investigate the reabsorptive steps of thiamin transport in brush border membrane vesicles (BBMVs) from rat kidney proximal tubules using tritiated thiamin with a high specific activity. METHODS: BBMVs prepared from the cortex kidney of rats were suspended in different media, controlling the composition of the intravesicular fluid by prolonged equilibration at 4 degrees C in preincubation buffers of desired composition. Then they were held on ice until used, when they were warmed at 25 degrees C for the uptake experiments. The amount of radioactivity taken up by the vesicles was measured radiometrically after separation with a rapid-filtration procedure. RESULTS: The time course profile of thiamin uptake was Na+ independent; 53% of thiamin taken up was membrane bound. The concentration curve had a biphasic course that was nonlinear (saturable) at physiological concentrations (<1.25 micromol/L) and linear at higher ones. Thiamin uptake was stimulated several-fold by an outwardly directed H+ gradient (pHin 6:pHout 7.5), which caused a transient accumulation of thiamin inside BBMVs against a concentration gradient. The enhanced thiamin uptake was only due to the H+ gradient, which made thiamin binding virtually negligible compared with translocation, and maintained the biphasic course of the concentration curve. The saturable component, however, had kinetic constants 12-fold higher than those in the absence of gradient. Moreover, the saturable component was inhibited by nonlabeled thiamin and its structural analogues, by inhibitors of intestinal thiamin/H+, renal guanidine/H+, and Na+/H+ antiporters, while it remained unmodified by some typical organic cations transported in renal BBMVs. CONCLUSION: The results provide strong evidence for the presence in renal BBMVs of a thiamin/H+ antiport having a 1:1 stoichiometric ratio. PMID- 10792624 TI - Cardiac infarcts increase sodium transporter transcripts (rBSC1) in the thick ascending limb of Henle. AB - BACKGROUND: Enhanced expression of the kidney-specific sodium transporter, rBSC1, in the thick ascending limb of Henle (TAL) and of the renal water channel, aquaporin-2 (AQP2), in collecting duct has been identified in rats with congestive heart failure (CHF) as a cause for enhanced sodium and water retention in this condition. However, the mechanism of impaired urinary sodium excretion observed even in rats with mild cardiac dysfunction remains unknown. METHODS: Male Sprague-Dawley rats with myocardial infarctions measuring 15 to 30% of the left ventricular circumference with no overt CHF were prepared. We measured the amount of rBSC1 or AQP2 mRNA using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of the PCR product for rBSC1 or by deleting 180 bp from the 760 bp PCR product for AQP2, respectively. The results were confirmed by in situ hybridization. rBSC1 protein expression was examined by immunohistochemistry and Western blot analysis using a specific antibody against rBSC1. RESULTS: Although plasma renin activity was slightly elevated in rats with myocardial infarction (MI), no significant differences in lung weight or plasma concentrations for aldosterone and atrial natriuretic peptide were observed between control rats and MI rats. Competitive PCR showed a significant increase in rBSC1 mRNA in the renal outer medulla and cortex of MI rats, which was confirmed by in situ hybridization. However, the AQP2 mRNA of these rats remained unchanged throughout the kidney. Renin-angiotensin II blockade by oral captopril administration did not influence the alteration in rBSC1 mRNA induced by myocardial infarction. Immunohistochemistry and Western blots showed the enhanced expression of rBSC1 protein in TAL of rats with small to moderate cardiac infarcts. CONCLUSIONS: rBSC1 is up-regulated even in rats with small to moderate myocardial infarctions, which may enhance the sodium transport in the TAL in this pathophysiologic condition. PMID- 10792625 TI - Effects of spironolactone on systolic blood pressure in experimental diabetic rats. AB - BACKGROUND: Mineralocorticoid hormones, which maintain electrolyte balance and blood pressure, are thought to be associated not only with the expression of renal 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), but also with that of intracellular mineralocorticoid receptors (MRs). The present study was designed to test whether the mineralocorticoid action of glucocorticoid corticosterone on renal MR is involved in the development of diabetes-associated hypertension by measuring the alterations of renal 11beta-HSD2. METHOD: We measured the mean systolic blood pressure, renal 11beta-HSD1, and mRNA levels in streptozotocin (STZ)-induced diabetic rats that received spironolactone, insulin, or no treatment, and in nondiabetic controls that received spironolactone. RESULTS: Four weeks after an injection of STZ, the renal 11beta-HSD2 and mRNA levels were significantly lower in diabetic rats than in control rats, and the mean systolic blood pressure was 14.8% higher in diabetic rats than in controls. Subcutaneous injections of spironolactone into diabetic rats for three weeks partially reversed the decrease in renal 11beta-HSD2 activity and gene expression, and prevented the mean systolic blood pressure elevation. Spironolactone treatment for one week also resulted in a significant reduction in mean systolic blood pressure during the development of diabetic hypertension. However, treatment with STZ did not significantly decrease the renal 11beta-HSD1 activity and mRNA expression, and spironolactone treatment did not exert a significant effect on this enzyme in STZ-induced diabetic rats. CONCLUSION: In the development of diabetes-induced hypertension, the effect of spironolactone on mean systolic blood pressure may be associated with the mineralocorticoid effects of corticosterone on renal MR, as well as an alteration of renal 11beta-HSD2 activity and its mRNA expression in insulin-dependent diabetic rats. PMID- 10792626 TI - Tobacco, hypertension, and vascular disease: risk factors for renal functional decline in an older population. AB - BACKGROUND: A decline in renal function with age has been noted in some but not all individuals. The purpose of this study was to identify risk factors associated with a clinically significant increase in serum creatinine (of at least 0.3 mg/dL) in an older nondiabetic population. METHODS: A retrospective case-control study was performed analyzing data obtained from 4142 nondiabetic participants of the Cardiovascular Health Study Cohort, all at least 65 years of age, who had two measurements of serum creatinine performed at least three years apart. Cases were identified as participants who developed an increase in serum creatinine of at least 0.3 mg/dL, with controls including participants who did not sustain such an increase. RESULTS: There was an increase in the serum creatinine of at least 0.3 mg/dL in 2.8% of the population. In a multivariate "best-fit" model adjusted for gender, weight, black race, baseline serum creatinine, and age, the following factors were associated with an increase in serum creatinine: number of cigarettes smoked per day, systolic blood pressure, and maximum internal carotid artery intimal thickness. CONCLUSIONS: These data suggest that three very preventable or treatable conditions-hypertension, smoking, and prevalent vascular disease, which are associated with large and small vessel disease-are highly associated with clinically important changes in renal function in an older population. PMID- 10792627 TI - Effects of furosemide on medullary oxygenation in younger and older subjects. AB - Renal medullary hypoxia is characteristic of mammalian kidneys and can be assessed noninvasively in animals and humans by blood oxygen level-dependent magnetic resonance imaging (BOLD MRI). Water diuresis has been shown to improve medullary oxygenation in young human subjects but not in elderly subjects, a difference attributed to a decline in renal prostaglandin production with age. Loop diuretics such as furosemide also increase medullary oxygenation in experimental animals, by inhibiting active transport and oxygen consumption in the medullary thick ascending limb. We examined, using BOLD MRI, this response to furosemide in eight younger (23 to 34 years) and eight elderly (64 to 81 years) healthy women. We also attempted to assess the role of prostaglandins in age related differences, using ibuprofen to inhibit prostaglandin E2 synthesis. Renal medullary oxygenation, initially low, increased during furosemide diuresis in younger subjects. In the older population, however, furosemide usually elicited little or no change in oxygenation of the renal medulla, despite profuse diuresis. Ibuprofen did not inhibit the action of furosemide to improve medullary pO2 in younger subjects. CONCLUSIONS: The action of loop diuretics to improve medullary oxygenation, apparent in younger subjects, is blunted by normal aging. Inhibition of prostaglandin synthesis did not counteract the effect of furosemide in younger subjects, suggesting that a decline in prostaglandin E2 production with age is not the central cause of this age-related defect. PMID- 10792628 TI - Glomerular type IV collagen in patients with diabetic nephropathy with and without additional glomerular disease. AB - BACKGROUND: Type IV collagen is a constituent of mesangial matrix and is increased in amount in many forms of glomerular injury. METHODS: We performed renal biopsies in patients who (1) were donating a kidney to a relative (LRD, N = 6), (2) had diabetic glomerulopathy with or without nephrosclerosis (DM, N = 6), or (3) had diabetic glomerulopathy with a superimposed glomerular lesion (DM+, N = 5). Glomerular collagen alpha2(IV) and control glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNAs were measured, and the former correlated with clinical and morphological data to assess its usefulness in reflecting glomerular injury. RESULTS: Collagen alpha2(IV) mRNA levels were lowest in LRD (2.9 +/- 0.6 attomol/glomerulus), higher in DM (5.9 +/- 1.6, P = 0.05), and highest in DM+ (12.7 +/- 2.8 attm/glomerulus, P < 0.05 vs. LRD and vs. DM). Control GAPDH mRNA levels were not significantly different (P > 0.05). Levels of proteinuria, serum creatinine, and glomerular size did not correlate with collagen alpha2(IV) mRNA levels. The fractional mesangial area and the fractional mesangial area occupied by type IV collagen were higher in both diabetic groups than in LRD (P < 10-6), but the intensity of type IV collagen staining in the diabetic patients was significantly less than that seen in the LRD (P < 0.01). In DM+ patients, extramesangial type IV collagen was present. Fractional mesangial area and glomerular collagen alpha2(IV) mRNA levels correlated (r = 0.45, P < 0.05). CONCLUSION: These data are consistent with a view of diabetic nephropathy as a lesion of increased alpha2 type IV collagen transcription, increased total amount of collagen present, but decreased mesangial density relative to other matrix molecules. These data further demonstrate that glomerular injury superimposed on diabetic nephropathy contributes to additional structural damage by inducing increased synthesis of type IV collagen at extramesangial sites. PMID- 10792629 TI - Multiple measurements of depression predict mortality in a longitudinal study of chronic hemodialysis outpatients. AB - BACKGROUND: The medical risk factors associated with increased mortality in hemodialysis (HD) patients are well known, but the psychosocial factors that may affect outcome have not been clearly defined. One key psychosocial factor, depression, has been considered a predictor of mortality, but previous studies have provided equivocal results regarding the association. We sought to determine whether depressive affect is associated with mortality in a longitudinal study of end-stage renal disease (ESRD) patients treated with HD, using multiple assessments over time. METHODS: Two hundred ninety-five outpatients with ESRD treated with HD were recruited from three outpatient dialysis units in Washington D.C. to participate in a prospective cohort study with longitudinal follow-up. Patients were assessed every six months for up to two years using the Beck Depression Inventory (BDI), age, serum albumin concentration, Kt/V, and protein catabolic rate (PCR). A severity index, previously demonstrated to be a mortality marker, was used to grade medical comorbidity. The type of dialyzer with which the patient was treated was noted. Patient mortality status was tracked for a minimum of 20 and a maximum of 60 months after the first interview. Cox proportional hazards models, treating depression scores as time-varying covariates in a univariable analysis, and controlling for age, medical comorbidity, albumin concentration, and dialyzer type and site in multivariable models, were used to assess the relative mortality risk. RESULTS: The mean (+/- SD) age of our population at initial interview was 54.6 +/- 14.1 years. The mean PCR was 1.06 +/- 0.27 g/kg/day, and the mean Kt/V was 1.2 +/- 0.4 at baseline, suggesting that the patients were well nourished and dialyzed comparably to contemporary U.S. patients. The patients' mean BDI at enrollment was 11.4 +/- 8.1, in the range of mild depression. Patients' baseline level of depression was not a significant predictor of mortality at 38.6 months of follow-up. In contrast, when depression was treated as a time-varying covariate based on periodic follow-up assessments, the level of depressive affect was significantly associated with mortality in both single variable and multivariable analyses. CONCLUSIONS: Higher levels of depressive affect in ESRD patients treated with HD are associated with increased mortality. The effects of depression on patient survival are of the same order of magnitude as medical risk factors. Our findings using both controls for factors possibly confounded with depressive affect in patients with ESRD and time-varying covariate analyses may explain the inconsistent results of previous studies of depression and mortality in ESRD patients. Time-varying analyses in longitudinal studies may add power to defining and sensitivity to establishing the association of psychosocial factors and survival in ESRD patients. The mechanism underlying the relationship of depression and survival and the effect of interventions to improve depression in HD outpatients and general medical inpatients should be studied. PMID- 10792630 TI - Impaired monocyte CD11b expression in interstitial inflammation in hemodialysis patients. AB - BACKGROUND: It is not known to what extent intravascular phenotypic alterations in adhesion molecule expression induced by hemodialysis influence the recruitment of monocytes and their ability to up-regulate CD11b at the local site of inflammation in the interstitium. Using a skin suction chamber technique, we addressed these issues in eight hemodialysis patients and in eight healthy subjects. METHODS: Two skin blisters were raised on the forearm of each individual and blister exudate collected. The blisters were then stimulated with autologous serum (active blister, intense inflammation) or buffer (control blister, intermediate inflammation), respectively. Thereafter the patients were treated with Cuprophan hemodialysis for four hours. After 10 hours, the exudate was aspirated from each chamber in all subjects. Monocyte count and expression of CD11b were analyzed in serum and blister fluid by flow cytometry. Then, monocytes from healthy blood donors were incubated in blister fluid from patients and healthy subjects in order to determine the local chemotactic activity in terms of CD11b up-regulation. Monocyte chemotactic protein-1 (MCP-1), a marker of systemic monocyte chemotactic activity, was also analyzed in serum at 0 and 10 hours in all individuals. RESULTS: The number of monocytes at the site of inflammation in the interstitium in hemodialysis patients correlated with the expression of CD11b on transmigrated cells (r = 0.78, P < 0.001). Monocytes collected in the active blister fluid of dialysis patients expressed equal levels of CD11b as cells collected from healthy subjects. By contrast, monocytes collected from the control blisters of patients expressed lower levels of CD11b than cells from healthy subjects (P < 0.01), despite equal interstitial biological activity of CD11b-mobilizing factors in blister fluid from patients and healthy subjects and the fact that patients had higher systemic chemotactic activity in terms of MCP-1 concentration in serum (P < 0.001). CONCLUSION: Monocytes from hemodialysis patients have the capacity to mobilize CD11b to the same extent as cells from healthy individuals at the inflammatory spot, but more intense stimuli are required for such actions, probably because of a transient refractoriness. PMID- 10792631 TI - Impact of dialyzer membrane selection on cellular responses in acute renal failure: a crossover study. AB - BACKGROUND: When acute renal failure (ARF) is severe enough to require dialysis, in-hospital mortality rates approach 60%. These alarming figures have been ascribed in part to advanced age and the high prevalence of comorbid conditions. In the past several years, a number of attempts have been made to investigate the impact of dialyzer membrane type on clinical outcomes. Unfortunately, to date, clinical studies addressing this question have reported conflicting results. METHODS: This crossover study examined the effect of dialyzer membrane type on cytokine synthesis by peripheral blood mononuclear cells (PBMCs), superoxide release by neutrophils, and apoptosis or programmed cell death of neutrophils in 24 patients with ARF requiring intermittent hemodialysis. The patients were assigned in an alternate order to a low-flux cellulose acetate (CA) or polysulfone (PS) dialyzer. After three consecutive dialysis sessions, patients were crossed over to the second dialyzer for three additional treatments. These cellular responses were measured upon dialyzer assignment and at the third and sixth dialysis session in relationship to the dialyzer type. RESULTS: The results of the study showed no impact of dialyzer biocompatibility on synthesis of tumor necrosis factor-alpha (TNF-alpha) or interleukin 10 (IL-10) by PBMCs, superoxide release by neutrophils, or neutrophil apoptosis. This held true regardless of the initial dialyzer assignment. Furthermore, there was no correlation between dialysis adequacy (measured by single-pool Kt/V) and postdialysis cellular responses. CONCLUSIONS: In summary, this study refines the question of biocompatibility by comparing a substituted cellulose rather than unsubstituted cellulose dialyzer to a PS dialyzer in the setting of ARF. The results failed to demonstrate a dialyzer advantage on the selected cellular responses. PMID- 10792632 TI - QTc dispersion increases during hemodialysis with low-calcium dialysate. AB - BACKGROUND: The risk of ventricular arrhythmias is known to increase during hemodialysis (HD) treatment, but the cause of this phenomenon has remained unidentified. QT dispersion (= QTmax - QTmin) reflects heterogeneity of cardiac repolarization, and increased dispersion is known to predispose the heart to ventricular arrhythmias and sudden cardiac death. METHODS: We studied the effect of dialysate calcium concentration on cardiac electrical stability during HD treatment in 23 end-stage renal disease patients. Three HD treatments were applied with dialysate Ca++ concentrations of 1.25 mmol/L (dCa++1.25), 1.5 mmol/L (dCa++1.5), and 1.75 mmol/L (dCa++1.75). The QTc interval and QTc dispersion were measured before and after the three sessions. RESULTS: With the dCa++1.5 and dCa++1.75 dialyses, serum Ca++ increased and the QTc interval remained stable (dCa++1.5) or decreased (dCa++1.75), but no significant change was noted in QTc dispersion. With dCa++1.25 HD, serum Ca++ decreased (1.24 +/- 0.11 vs. 1.20 +/- 0.09 mmol/L, P < 0. 05), and both the QTc interval (403 +/- 27 vs. 419 +/- 33 ms, P < 0. 05) and QTc dispersion increased (38 +/- 19 vs. 49 +/- 18 ms, P < 0. 05). The change in the QTc interval correlated inversely with the change in serum Ca++ (r = -0.68, P < 0.0001). Except for serum Ca++ and plasma intact parathyroid hormone, predialysis and postdialysis values in other blood chemistry, blood pressure, heart rate, body weight, and total ultrafiltration were equal in the three dialysis sessions. CONCLUSION: This study is the first, to our knowledge, to demonstrate that HD increases QTc dispersion if a low-calcium (dCa++1.25) dialysate is used. This indicates that the use of low-calcium dialysate may predispose HD patients to ventricular arrhythmias and that perhaps it should be avoided, at least when treating patients with pre-existing cardiac disease. PMID- 10792633 TI - Glucose infusion test: a new screening test for vascular access recirculation. AB - BACKGROUND: Vascular access recirculation is an important cause of diminished dialysis efficiency. We propose a new screening test based on glucose infusion as a tracer for recirculation. METHODS: The glucose infusion test (GIT) protocol comprises a basal blood sample (A) from the arterial port, a 5 mL bolus of 20% glucose into the venous chamber (time 0), followed by a second sample (B) in four seconds (from 13 to 17 s with QB 300 mL/min) from the same port. The blood glucose level is determined at the bedside on A and B with a reflectance photometer (CV 1.8%). Interpretation of the test is straightforward: If B = A, there is no recirculation, whereas if B > A, recirculation can be calculated from the regression equation: 0.046 x (B - A) + 0.07, obtained from in vitro tests reproducing artificial recirculation at 0, 5, and 10%. To validate this new method in vivo, we compared GIT and the urea test on 39 hemodialysis patients, obtaining a good correlation (r = 0.93). The two tests were considered positive (recirculation present) when the lower 95% confidence intervals were more than zero. RESULTS: Our patients were divided into two groups: those with (22 out of 39, mean recirculation 11.8%) or without recirculation (17 out of 39, mean 0.06%). The urea test did not recognize 7 out of 22 patients because they had a small recirculation below the urea test limit of detection. CONCLUSIONS: GIT was more sensitive (detection limit 0.3%), simpler, and immediate in showing the results than the urea test. It is an accurate and low-cost technique for screening and follow-up of vascular access in a dialysis unit. PMID- 10792634 TI - Simultaneous pancreas-kidney transplantation reduces excess mortality in type 1 diabetic patients with end-stage renal disease. AB - BACKGROUND: Diabetic renal disease continues to be the most significant cause of end-stage renal disease (ESRD) in the United States. Renal transplantation improves diabetic ESRD patient survival; however, the diabetic state remains associated with poor patient survival. Simultaneous pancreas-kidney (SPK) transplantation can restore normoglycemia and thus may improve outcomes. METHODS: We assessed the impact of SPK on age-range-matched type 1 diabetic patients who underwent renal transplantation at a single center. The observed/expected life span and annual mortality rates (AMRs) were used as measures of survival. A Cox proportional hazards analysis was used to analyze the impact of potential variables on mortality in SPK recipients. RESULTS: SPK transplantation (N = 335) increased the observed/expected life span compared with diabetic cadaveric (DM Cad, N = 147) and live-donor (DM-Live, N = 160) transplant recipients (P = 0.004) and significantly reduced the AMRs (SPK, 1. 5%; DM-Cad, 6.27%; DM-Live, 3.65%, P = 0.008, SPK vs. other DM). Moreover, the SPK observed/expected life span and AMR were not significantly different from that of age-range-matched nondiabetic transplant recipients (N = 492). The only variable that was significantly associated with patient survival was discharge serum creatinine (relative risk 1.16, P < or = 0.0154). CONCLUSION: These data demonstrate that SPK improves the ability for type 1 diabetic patients to live more of their expected life span. This suggests that glycemic control, even as a late intervention in a diabetic patient's lifetime, may beneficially affect survival. PMID- 10792635 TI - Outcomes of renal transplantation among patients with end-stage renal disease caused by lupus nephritis. AB - BACKGROUND: Although the outcomes of renal transplantation among patients with end-stage renal disease (ESRD) caused by lupus nephritis have generally been found to be comparable to those of patients with other causes of ESRD, some studies indicate that cadaveric graft failure is more common among these patients. However, most previous studies examined small numbers of patients and did not adjust for important confounding factors. METHODS: Graft failure and patient mortality after the first cadaveric renal transplantation were compared between 772 adults with ESRD caused by lupus nephritis and 32,644 adults with ESRD caused by other causes who received a transplant between 1987 and 1994 and were included in the United States Renal Data System. The median follow-up times were 4.9 and 5.0 years in the two groups, respectively. Multivariate Cox regression models were used to adjust the risks of graft failure and mortality for group differences in recipient and donor characteristics. Similar comparisons were performed between 390 adults with ESRD caused by lupus nephritis and 10,512 adults with ESRD caused by other causes after first living-related renal transplantation. RESULTS: In an unadjusted analysis, the risk of graft failure after first cadaveric transplant was slightly but significantly greater among patients with ESRD caused by lupus nephritis than among those with ESRD caused by other causes [hazard ratio (HR), 1.13; 95% CI, 1.01 to 1. 26, P = 0.04]. However, after adjustment for potential confounding factors, the risk of graft failure was not increased in patients with ESRD caused by lupus nephritis (HR, 1.08; 95% CI, 0.94 to 1.23, P = 0.28). Mortality after the first cadaveric transplantation did not differ between groups. The adjusted risks of graft failure (HR, 1.06; 95% CI, 0.84 to 1.32, P = 0.62) and patient mortality (HR = 0. 69; 95% CI, 0.45 to 1.05, P = 0.09) after the first living-related renal transplant were also not significantly higher among patients with ESRD caused by lupus nephritis. CONCLUSIONS: Graft and patient survival after first cadaveric and first living related renal transplants are similar in patients with ESRD caused by lupus nephritis and patients with ESRD from other causes. PMID- 10792636 TI - Predicting clinical outcome in the elderly renal transplant recipient. AB - BACKGROUND: The purpose of this study was to evaluate graft and patient survival in first-time kidney transplant recipients 60 years old or older, and to identify pretransplant risk factors that predict clinical outcome. METHODS: We reviewed the clinical course of 206 recipients, 60 years old or older, of first kidney transplants at the University of Minnesota. Patient and graft survival were compared with 1640 patients aged 18 to 59 transplanted during the same time period. Regression analysis was performed to identify risk factors that predicted a poor outcome. RESULTS: In patients 60 years old or older, graft survival at one and five years was 86 and 60%, and patient survival at one and five years was 90 and 68%, respectively. Graft and patient survival were decreased compared with recipients aged 18 to 59, but were similar when censored for patient death as a cause of graft loss. A pretransplant history of nonskin malignancy and vascular disease and a current smoking history were risk factors for decreased graft and patient survival. To determine the potential impact of screening for low-risk patients, we evaluated graft and patient survival in patients age > or =60 without these risk factors versus those with one or more risk factors. In the absence of risk factors, both graft and patient survival were significantly improved compared with patients with these risk factors and were equivalent to that of patients aged 18 to 59. CONCLUSIONS: Renal transplantation is a safe and effective therapy for the older renal failure patient. In the absence of identified risk factors, graft survival is equivalent to that seen in younger patients. PMID- 10792637 TI - Bacteremia associated with tunneled dialysis catheters: comparison of two treatment strategies. AB - BACKGROUND: Tunneled dialysis catheters are often used for temporary vascular access in hemodialysis patients, but are complicated by frequent systemic infections. The treatment of bacteremia associated with infected tunneled catheters requires both antibiotic therapy and catheter replacement. We compared the outcomes of two treatment strategies for catheter-associated bacteremia: exchange of the existing catheter with a new one over a guidewire versus catheter removal with delayed replacement. METHODS: We retrospectively analyzed the outcomes of all cases of tunneled dialysis catheter-associated bacteremia during a two-year period. The infection-free survival time of the subsequent catheter was evaluated in two groups of patients: group A (31 catheters), exchange of the existing infected catheter with a new catheter over a guidewire, and group B (38 catheters), removal of the infected catheter followed by delayed catheter replacement 3 to 10 days later. Patients in both groups received three weeks of systemic antibiotic therapy. Cox proportional hazard models were used to evaluate the factors predictive of infection-free survival time of the replacement catheter. RESULTS: On univariate proportional hazard regression analysis, the infection-free survival time of the replacement catheter was similar for groups A and B (P = 0.72), whereas the hazard of infection was significantly greater for patients with hypoalbuminemia (serum albumin < 3.5 g/dL), as compared with patients with a normal serum albumin (hazard ratio 2.81, 95% CI, 1. 21, 6.53, P = 0.016). The infection-free survival time was not affected by patient age, sex, diabetic status, or type of organism (gram-positive coccus vs. gram-negative rod). CONCLUSIONS: The infection-free survival time associated with the subsequent catheter is similar for the two treatment strategies. However, exchanging the catheter for a new one over a guidewire minimizes the number of separate procedures required by the patient. Hypoalbuminemia is the major risk factor for recurrent bacteremia in the replacement catheter. PMID- 10792638 TI - Franz Volhard and his students' tortuous road to renovascular hypertension. AB - Franz Volhard and his students' tortuous road to renovascular hypertension. Harry Goldblatt's name is irrevocably linked to the phenomenon that renal artery constriction increases blood pressure via renin release, even in the absence of significantly decreased renal function. However, "getting there was more than half the fun." A lively competition took place earlier in this century elucidating the role of the kidney in hypertension, in which Franz Volhard and his disciples played a major role. I have reviewed the extensive German literature of the time, and observed that Franz Volhard and his young associates made major contributions investigating decreases in renal perfusion and resultant increases in blood pressure. Hessel, Hartwich, and Volhard made seminal observations in this regard that even preceded those of Goldblatt. A sojourn into this past history gives a revealing insight into our cumulative-albeit still incomplete-knowledge of the kidney, renin-angiotensin system, and blood pressure regulation. Franz Volhard was a colorful figure whose contributions extended far beyond classifying renal diseases. PMID- 10792639 TI - Development of vascular elements during renal organogenesis. PMID- 10792640 TI - Prospects for gene transfer into renal cells. PMID- 10792641 TI - Proteinase 3: A cofactor for the binding of antineutrophil cytoplasm antibodies (ANCA) to endothelial cells? PMID- 10792642 TI - Kidney biopsy and patient outcome. PMID- 10792643 TI - Laparoscopic donor nephrectomy. PMID- 10792644 TI - Probeliatrade mark PCR system for rapid detection of Salmonella in milk powder and ricotta cheese. AB - The Probeliatrade mark Salmonella sp. PCR amplification and detection kits (Sanofi Diagnostics Pasteur, Marnes La Coquette, France) were evaluated for the rapid and specific detection of Salmonella agona artificially inoculated into skim milk powder and ricotta cheese. The Probeliatrade mark results were compared with those obtained using the Australian Standard Method. Using a pure culture of Salm. agona, the detection limit of Probeliatrade mark was between 8 and 79 cfu ml-1, equivalent to 0.2-2 cfu per PCR reaction. Detection of Salm. agona inoculated in skim milk powder (at 5-10 cfu g-1, stored at 5, 15 or 25 degrees C) and ricotta cheese (at 1-2, 10-20 and 100-200 cfu per 25 g) was effected by using non-selective enrichment prior to the PCR determinations. For all of the 40 milk powder samples and 12 ricotta cheese samples, the Probeliatrade mark results were consistent with those using the Australian Standard Method. Using Probeliatrade mark, Salmonella was detected to genus level in the dairy products within 24-28 h, whereas the cultural technique required 3-4 d for presumptive positive isolates and further time for confirmation. PMID- 10792645 TI - Influence of the growth rate calculation on the relationship between growth rate and temperature. AB - Three calculations of the growth rate (e.g. slope of a plot of the log10 of cfu ml-1 vs time, mum of the Gompertz equation and the reciprocal of time to obtain 108 cfu ml-1) were compared for Escherichia coli TG1 growing in tryptone soy broth medium at temperatures ranging from 14 to 39 degrees C. Up to now, the influence of using such different definitions on the relationship between microbial growth rate and temperature has never been investigated. In order to compare these calculation procedures, a dimensionless analysis based on the following normalized variables, mudim = mu/muopt and Tdim = [T-Tmin]/[Topt-Tmin], was used (Dantigny 1998). The influence of suboptimal temperatures on the growth rate was represented by means of a Belehradek-type model based on a power function law: [mudim] = [Tdim]alpha. The influence of the different growth rate calculations on the model constants was assessed. Despite the great dependence of the raw growth rate values on the calculation procedure, the dimensionless analysis demonstrated that the alpha-value is independent of the growth rate definition. This result suggests that any definition for the growth rate can be utilized in studies aimed at determining the influence of temperature on microbial growth and highlights the interest of using dimensionless variables to overcome differences in the order of magnitude of the growth rate data and to avoid confusion between definitions. PMID- 10792646 TI - Calmodulin mediated activation of acetyl-CoA carboxylase during aflatoxin production by Aspergillus parasiticus. AB - The relevance of Ca2+-calmodulin-mediated processes in channelling acetate for aflatoxin formation was investigated by studying the influence of trifluoperazine (an anticalmodulin agent) on [14C]-acetate incorporation and activity of acetyl CoA carboxylase in Aspergillus parasiticus NRRL 2999. Culturing the organism in presence of 0.14 mmol l-1 trifluoperazine resulted in 55% decrease of [14C] acetate incorporation into aflatoxin B1, along with an 80% decrease in acetyl-CoA carboxylase activity at periods corresponding to maximal aflatoxin production. Concomitant decrement (35%) in the activity of glucose-6-phosphate dehydrogenase indicated decreased availability of reduction potential (NADPH) required for aflatoxin biosynthesis. The ability of calmodulin to activate and trifluoperazine to inhibit acetyl-CoA carboxylase activity in a dose-dependent manner was also noted under in vitro conditions. The combined results suggest calmodulin-mediated activation of acetyl-CoA carboxylase as an important event for aflatoxin production. PMID- 10792647 TI - Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment. AB - Intact conidia of three industrially relevant strains of Trichoderma reesei were effectively transformed by particle bombardment. Transformations were carried out individually with plasmids carrying either the fungal amdS or bacterial hph gene as a selectable marker and by cotransformation with both plasmids. Transformant yields with single plasmids were up to 11 stable transformants per microg DNA at the bombardment distance of 6 cm. Mitotic stability of the transformants was 75 100% and the cotransformation efficiency averaged 92% when the first selection was performed on hygromycin B plates. The entire procedure could be completed in 1 week with the hph marker. PMID- 10792648 TI - Genetic polymorphism in Xanthomonas albilineans strains originating from 11 geographical locations, revealed by two DNA probes. AB - Two DNA fragments from Xanthomonas albilineans were used as probes to study the molecular diversity among strains of this pathogen. Two serologically distinct groups, serovars I and II, could be differentiated by hybridization to the probes. These probes, designated 830 and 838, were cloned after subtractive DNA hybridization of common sequences of Xanthomonas campestris pv. vasculorum from a serovar I strain of X. albilineans. They did not hybridize to the DNA of several other xanthomonads or to sugarcane DNA under the conditions of hybridization used. Faint bands were observed upon hybridization of probe 830 with one strain of X. campestris pv. phaseoli. The same banding patterns were obtained with a strain of X. albilineans from Burkina Faso and the serovar II strains of Mauritius. The serovar I strains from Mauritius and two other strains each from Reunion and South Africa had similar pattern. PMID- 10792649 TI - The outer membrane of Pseudomonas aeruginosa NCTC 6749 contributes to its tolerance to the essential oil of Melaleuca alternifolia (tea tree oil). AB - Pseudomonas aeruginosa is less susceptible to the antimicrobial properties of tea tree oil than many bacteria and its tolerance is considered to be due to its outer membrane. Polymyxin B nonapeptide (PMBN), which has no antibacterial action, was used to permeabilize the outer membrane. The addition of PMBN to Ps. aeruginosa NCTC 6749 markedly increased this organism's susceptibility to tea tree oil and to its normally inert hydrocarbons, p-cymene and gamma-terpinene. PMID- 10792650 TI - The solubility of the ribotoxin alpha-sarcin, produced as a recombinant protein in Escherichia coli, is increased in the presence of thioredoxin. AB - The yield of purified recombinant alpha-sarcin increases approximately three- to fourfold when this toxin is co-expressed in Escherichia coli with thioredoxin. This increased production is attributed to the existence, in the presence of thioredoxin, of a reducing environment which allows rearrangement of incorrect disulphide bonds to produce the soluble native conformation. The protein thus produced retains the structural, spectroscopic and enzymatic features of the natural fungal alpha-sarcin. PMID- 10792651 TI - An evaluation of a laser scanning device for the detection of Cryptosporidium parvum in treated water samples. AB - A laser scanning device, the ChemScan RDI (Chemunex, Paris, France), was compared with manual fluorescence microscopy for the detection of oocysts of Cryptosporidium. Pairs of filters were spiked with approximately 100 oocysts. Over 24 h at least 1000 l of treated water was passed through the filters, then concentrated deposits were subjected to an immunomagnetic separation (IMS) protocol described by the manufacturer (Dynal, Oslo, Norway) and examination by fluorescence microscopy, or an IMS protocol (Chemunex) and detection by ChemScan laser scanning. Subsequently a set of five 1-ml samples containing oocysts over a range of concentrations, including a negative control, were examined blind by the two methods (stage two). In stage 1 the average recovery rates were estimated to be 49% (manual fluorescence microscopy) and 73% (ChemScan). The average ratio of ChemScan to manual fluorescence microscopy counts was 1.54 (range 1.08-2.36). In stage 2, statistical comparison of all but one set of results showed there was no significant difference between methods. Differences for the high count sample may possibly have been caused by duplicate counting of oocysts by manual fluorescence microscopy. PMID- 10792652 TI - Adherence of Fusobacterium necrophorum subsp. necrophorum to ruminal cells derived from bovine rumenitis. AB - Fusobacterium necrophorum subsp. necrophorum strain VPI 2891 was shown to adhere to the surfaces of ruminal cells derived from bovine rumenitis. The strain also attached to bovine type 1 collagen. Treatment of the bacterium with antiserum to bacterial cells reduced attachment. The bacterial attachment was also markedly reduced when the ruminal cells had been pretreated with anticollagen serum. Fluorescence specific for the collagen was demonstrated on the surface of bovine tissue affected with rumenitis. These findings suggest that F. necrophorum subsp. necrophorum strain VPI 2891 adheres to the ruminal cells derived from rumenitis tissue and that the attachment may be mediated by cellular collagen. PMID- 10792653 TI - Rapid detection of malto-oligosaccharide-forming bacterial amylases by high performance anion-exchange chromatography. AB - High performance anion-exchange chromatography with pulsed amperometric detection was applied for the rapid analysis of malto-oligosaccharides formed by extracellular enzyme preparations from 49 starch-degrading bacterial strains isolated from soil and compost samples. Malto-oligosaccharide-forming amylases, indicated by a predominant formation of maltohexaose from starch, were produced by enzyme preparations from four of the isolates growing at pH 7.0 and 10. PMID- 10792655 TI - Comparison of methods for the recovery and detection of low levels of injured Salmonella in ice cream and milk powder. AB - This study compared the ability of four rapid methods and a standard cultural method to detect low levels of heat-injured cells of Salmonella typhimurium in ice cream and skimmed milk powder. The detection of Salmonella in samples contaminated with low levels (< 10 cfu 25 g-1) was significantly greater with the novel broth method than with the other methods (P 10 cfu 25 g-1, there was no significant difference between the methods except for the novel broth method and a dipstick-based immunoassay (P /=12 mm) were assayed for the minimum inhibitory concentration and submitted to phytochemical screening by thin-layer chromatography and bioautography. The results obtained indicate that the diethyl ether extracts were the most efficient antimicrobial compounds. The activity was more pronounced against Gram-positive and fungal organisms than against Gram negative bacteria. Bioautography showed that the antimicrobial activity was probably due to flavonoids and terpenes. PMID- 10792668 TI - Effect of ammonia on the growth of Bacillus species and some other bacteria. AB - The tolerance of 26 Bacillus species isolated from alkaline fermented foods, five other bacilli and nine non spore-forming bacteria to alkaline pH and ammonia was determined. All grew at pH 7, 8 and 9 in the presence of 930 mmol l-1 NH4 + at pH 7.0, and in the presence of NH3 concentrations up to 5 mmol l-1 at pH 7.0 and 8.0. At higher NH3 concentrations, growth of some of the bacteria was inhibited and at 500 mmol l-1 only B. pasteurii and B. pumilus grew. Bacteria from alkaline food fermentations included strains relatively sensitive to NH3 (inhibited by 50 mmol l-1) and relatively tolerant strains (grew in the presence of 300 mmol l-1) and there was no evidence that they were more tolerant to NH3 than bacteria not associated with these fermentations. PMID- 10792669 TI - The relationship between hide cleanliness and bacterial numbers on beef carcasses at a commercial abattoir. AB - Cattle were visually inspected in the lairage of a commercial abattoir and assigned to a category ranging from 1 (very clean) to 5 (very dirty) depending on the observed cleanliness of the hide. Animals from categories 2, 3 and 5 were slaughtered and total viable counts (TVCs) enumerated at five sites (hock, brisket, cranial back, bung and inside round) on the subsequent carcasses. The TVCs at the hock were significantly higher on category 5 than on category 2 carcasses (P < 0.05). Similarly, TVCs at the brisket were significantly higher on categories 3 and 5 than on category 2 carcasses (P < 0.05). There were no differences in counts among the categories at any of the other sites. The TVCs averaged over the five carcass sites were higher for category 5 than for category 2 carcasses (P < 0.05). The TVCs at the brisket were significantly higher than all other sites (P < 0.01). In general, carcasses from category 2 animals slaughtered in a batch with dirtier animals (categories 3 and 5) did not have higher TVCs than carcasses of category 2 animals slaughtered at the beginning of the day in the absence of dirtier animals. The introduction of improved hygienic practices during the dehiding of category 4 animals resulted in reduced TVCs at the brisket (P < 0.001). PMID- 10792670 TI - Microbial degradation and detoxification of high molecular weight polycyclic aromatic hydrocarbons by Stenotrophomonas maltophilia strain VUN 10,003. AB - The ability of Stenotrophomonas maltophilia strain VUN 10,003 to degrade and detoxify high molecular weight polycyclic aromatic hydrocarbons (PAHs) was evaluated in a basal liquid medium. Using high cell density inocula of strain VUN 10,003, the concentration of pyrene, fluoranthene, benz[a]anthracene, benzo[a]pyrene, dibenz[a, h]anthracene and coronene decreased by 98, 45, 26, 22, 22 and 55% over periods ranging from 5 to 42 d. When a PAH mixture containing three- to seven-ring compounds was used, degradation of both low and high molecular weight compounds occurred concurrently. Mutagenicity assays (Ames Test) demonstrated a decrease in the mutagenic potential of dichloromethane culture extracts from all cultures containing single PAH over the incubation period, corresponding to the decrease in the concentration of the PAH. These observations indicate that strain VUN 10,003 could be used for the detoxification of PAH contaminated wastes. PMID- 10792671 TI - Treatment of sanitary-important bacteria by bacteriocin substance V24 in cattle dung water. AB - Quantification of sanitary-important bacteria (e.g. Enterobacteriaceae), as well as indicators of environmental contamination, was assessed in samples of cattle dung from 25 cattle farms in 15 north-eastern Slovakia districts. The inhibitory effect of crude bacteriocin extract CBE V24 from Enterococcus faecalis V24 against Listeria monocytogenes Ohio and Yersinia enterocolitica YE85 was examined in cattle dung water with the aim of finding a new way of eliminating the health risk of the animal slurry. The following bacterial groups were quantified: Salmonella spp., Shigella-like spp. , Proteus spp., Enterobacter spp., Citrobacter spp., Pseudomonas spp. , Escherichia coli, Listeria spp., staphylococci, streptococci and enterococci (the average count ranged from 102 up to 104 cfu ml-1). Antagonistic effect of the crude bacteriocin from Enterococcus faecalis V24 in the range of 100-600 Arbitrary units per ml (AU ml-1) was shown against the following bacteria: Enterobacter cloacae, Ent. asburiae, Proteus spp., Salmonella spp., Acinetobacter lwoffi, L. monocytogenes as well as Y. enterocolitica YE85. During tests performed to study the inhibitory effect of the crude bacteriocin CBE V24 (concentration 800, 1600 AU ml-1) against L. monocytogenes Ohio and Y. enterocolitica YE85 in experimentally contaminated cattle dung, a reduction of 2.03 and 1.44 log cfu ml-1, respectively, was already noted after 1 h after crude bacteriocin CBE V24 addition. PMID- 10792672 TI - Use of Agrobacterium expressing green fluorescent protein to evaluate colonization of sonication-assisted Agrobacterium-mediated transformation-treated soybean cotyledons. AB - Colonization and infection of soybean cotyledons by Agrobacterium tumefaciens and subsequent elimination of bacteria from cotyledons were monitored using bacteria expressing green fluorescent protein (GFP). GFP provided a quick, non-destructive method to evaluate, in real time, Agrobacterium colonization of cotyledon surfaces as well as infection of internal cells. GFP was first detected 7 h following inoculation of the cotyledon. By 36 h, GFP expression was very intense, and was limited to the adaxial surface of the cotyledon. Expression of GFP also served as a useful indicator of successful elimination of the bacterium from plant tissue following antibiotic treatment. PMID- 10792673 TI - Use of 8-hydroxyquinoline-beta-D-glucuronide for presumptive identification of Shiga toxin-producing Escherichia coli O157. AB - 8-hydroxyquinoline-beta-D-glucuronide (HQG) was used to improve the presumptive identification of Shiga toxin-producing Escherichia coli O157 (STEC O157) on sorbitol MacConkey agars (SMAC). Advantages of HQG are (i) that it is less expensive than 5-bromo-4-chloro-3-indoxyl-glucuronide; (ii) that it is visible in normal daylight and (iii) that it does not diffuse into the agar like 4 methylumbelliferryl-beta-D-glucuronide (MUG). Sixteen STEC O157 isolates, 91 bovine mastitis-associated E. coli isolates and 222 faecal E. coli isolates from apparently healthy cattle were used in this study. 4-methylumbelliferryl-beta-D glucuronide detected beta-glucuronidase activity in more isolates than HQG (P < 0.05). On SMAC with HQG, cefixime and tellurite all STEC O157 isolates grew as cream-coloured colonies (100% sensitivity), whereas all non-STEC O157 E. coli except one grew either not at all or as purple or black colonies (99.7% specificity). No difference was found between faecal and mastitis isolates for the proportion of isolates that hydrolysed HQG or MUG or fermented sorbitol. However, significantly more mastitis isolates were able to grow in the presence of the cefixime-tellurite supplement. 8-Hydroxyquinoline-beta-D-glucuronide is a useful substrate for the identification of STEC O157 on SMAC. PMID- 10792674 TI - Citrate can partially replace carbon dioxide required for growth of Lactococcus lactis subsp. lactis biovar diacetylactis. AB - Lactococcus lactis subsp. lactis biovar diacetylactis was grown as batch cultures on a chemically defined medium. No growth was observed when the cultures were sparged with pure nitrogen (1.3 l l-1 min-1) whereas the cultures displayed exponential growth in the presence of minute amounts of carbon dioxide (0.035 mol % of the inlet gas). However, in the former case, the addition of citrate restored growth. This suggested that oxaloacetate required for aspartate biosynthesis can be formed by the carboxylation of pyruvate or by citrate catabolism. When the cultures were heavily sparged with nitrogen (2.6 l l-1 min 1), no growth was observed even in the presence of citrate. This indicated that growth in these conditions was repressed by the absence of carbon dioxide required in some other biosynthetic reaction than in the carboxylation of pyruvate leading to oxaloacetate/aspartate biosynthesis. PMID- 10792676 TI - Taking stock. PMID- 10792675 TI - RAPD-fingerprinting of Serratia marcescens after formaldehyde inactivation of DNase activity. AB - Serratia marcescens strains commonly cause hospital outbreaks. A random amplified polymorphic DNA (RAPD) assay was applied to the epidemiology of Ser. marcescens using formaldehyde fixation and boiling of bacterial cells for DNA extraction. The method preserved bacterial DNA and gave optimal results of RAPD fingerprinting, facilitating the investigation of hospital infections caused by Ser. marcescens. PMID- 10792677 TI - Are scientific papers out of date? PMID- 10792678 TI - Continuing education and professional development for hospital doctors and dentists. PMID- 10792679 TI - The value of collaboration by medical schools: lessons from the united states. PMID- 10792680 TI - What is different about medicine? PMID- 10792681 TI - What use are ethical codes? An analysis of three possible rationales for the use of ethical codes in medical schools and a review of the evidence relating to them. PMID- 10792682 TI - Continuing medical education and primary physicians' job stress, burnout and dissatisfaction. AB - OBJECTIVES: The association between opportunities for continuing medical education (CME) and primary physicians' job stress, burnout and job dissatisfaction has not been investigated. It was hypothesized that participation in CME activities and perceived opportunities at work for keeping up-to-date with medical and professional developments would be correlated negatively with job stress and burnout, and positively with job satisfaction. METHOD: 309 primary care physicians (183 family physicians and 126 paediatricians) employed in health maintenance organizations in Israel responded to a mailed questionnaire. The independent variables were the extent of engagement in CME activities and perceived opportunities at work for professional updating. The dependent variables were job stress, burnout and job satisfaction. Multiple regression analyses were employed. RESULTS: After controlling for age, sex and professional status, participation in CME activities was associated negatively with job stress and positively with job satisfaction, among family physicians. Among paediatricians, CME was associated negatively with burnout. Perceived opportunities for professional updating were associated negatively with burnout and (marginally) with stress among family physicians, and negatively with stress and positively with satisfaction among paediatricians. A third of the paediatricians and a quarter of the family physicians wanted to increase their involvement in CME. DISCUSSION: In this cross-sectional study, causality cannot be established and the CME measures should be refined. However, the results are consistent with the study's hypotheses and suggest that opportunities for CME and professional updating may reduce physicians' job distress and dissatisfaction. PMID- 10792683 TI - Use of scheme-based problem solving: an evaluation of the implementation and utilization of schemes in a clinical presentation curriculum. AB - CONTEXT: The University of Calgary has implemented a new curriculum which is organized according to 120 ways in which patients may present to a physician. Students are taught scheme-based problem solving rather than the more typical hypothetico-deductive or search and scan approach to problem resolution. OBJECTIVE: This study sought to determine the extent to which faculty and students were implementing and utilizing scheme-based problem solving. METHOD: All classes taught within the new clinical presentation curriculum were surveyed at the year end. Participants included four classes of first-year students and three classes of second-year students. Using a 5-point scale, students responded to survey items regarding scheme implementation and utilization. RESULTS: Data were analysed using MANOVA (multivariate analysis of variance) and revealed significant differences among classes in both first- and second-year students. Increments in scheme implementation and utilization by instructors and students were observed, although instructors' utilization of schemes lagged behind that of students. A levelling effect to the benefits of schemes for problem solving was also evident. First-year students reported schemes to be very useful for learning and organizing new information. CONCLUSION: Although it has taken time to implement curriculum change, the student response to schemes has been favourable. Faculty development and further generation of pictorial/spatial representations for all schemes, to ensure that all clinical presentations provide pathways that students can use for both learning and problem solving are recommended. Whether students who utilize schemes are more successful problem solvers is not yet known but will be the subject of study as soon as scheme delivery is predominant. PMID- 10792684 TI - Role modelling: how does it influence teaching in Family Medicine? AB - OBJECTIVE: To undertake a qualitative study to explore the influence of role modelling on teaching by comparing faculty members recollections of their teachers' behaviours with residents perceptions of the same behaviours in a family medicine residency programme in Saudi Arabia. METHOD: Using semi structured interviews of faculty and a questionnaire based on the issues arising from the interviews, faculty members' recollections of their medical teachers' behaviours were compared with residents' current perceptions of the same teaching behaviours. SETTING: Department of Family Medicine, King Fahad National Guard Hospital, Riyadh, Saudi Arabia. SUBJECTS: Faculty and residents. RESULTS: The four best-remembered teacher behaviours were: positive behaviour towards patients, negative behaviour towards junior colleagues, effective presentation of subject content and encouragement to participate in patient care. The residents perceived positive behaviour towards patients, positive behaviour towards junior colleagues, suboptimal skills of subject content presentation, and insufficient encouragement for trainees to actively participate in patient management. Although faculty retained many unhappy memories of teacher behaviour, it was encouraging that there was no evidence of perpetuation of the negatively perceived behaviours which provoked them. CONCLUSIONS: Discernment of the value of technical teaching skills was not a predictor of later proficiency. PMID- 10792685 TI - The Consortium of Graduate Medical Schools in Australia: formal and informal collaboration in medical education. AB - CONTEXT: In 1996-97 three Australian medical schools adopted 4-year graduate medical courses. The schools formed a consortium to establish common admissions processes and to collaborate on curriculum and staff development. OUTCOMES: ADMISSIONS: Outcomes in admissions included the construction of an entry test and agreement on common application procedures. Structured interviews and measures of prior tertiary performance were adopted in each school with some local variations. Formal structures were put into place to manage the development of the test and admissions procedures. OUTCOMES: CURRICULUM AND STAFF DEVELOPMENT: No formal structures were put into place for curriculum and staff development. While the advantages of common approaches were recognised, it was accepted that it was also important to generate local ownership and commitment to the new courses. Outcomes of educational consortia should not be judged solely by the nature of joint ventures but by the way in which membership of the consortium changes the priorities and practices within the institutions. Examples of the operation of this principle within the three schools include: use of visiting experts to refine and modify approaches to problem-based learning; use of staff from partner institutions to facilitate implementation of the courses; sharing information technology expertise; cooperation in the construction of feedback mechanisms, and sharing advice on fulfilling accreditation requirements. CONCLUSION: The Consortium has had an important impact on Australian medical education. There is a need for further analysis of the two methods of operation of the consortium, formal and informal, and their relation to its overall functioning. PMID- 10792686 TI - Medical education - beyond tomorrow? The new doctor - Asclepiad or Logiatros? AB - Against a background of the theoretical basis for the contextual approach to medical education, this paper examines and supports the changes that are occurring in undergraduate medical education throughout the world, before putting up for discussion the suggestion that the changes have not gone far enough. Consideration is given to a model of apprenticeship learning within undergraduate medical education, to the benefits it may offer, and to some of the challenges inherent in its implementation. PMID- 10792687 TI - An evaluation study of the didactic quality of clerkships. AB - CONTEXT: Previous qualitative research at the University of Antwerp revealed dissatisfaction amongst medical students about clinical clerkships. OBJECTIVE: To use quantitative methods to further explore student perceptions of their clinical teaching. METHOD, SAMPLE AND SETTING: Monthly questionnaire administered to final year medical students at the University of Antwerp, Belgium. RESULTS: The response rate was 83.9%. Many of the qualitative findings were reproduced. Educational resources were not optimally used. The junior doctor was the most important clinical teacher. Many activities were passive experiences. Coaching, feedback and supervision associated most with general satisfaction of the clerkship, however, these dimensions were often considered suboptimal. CONCLUSIONS: Clinical clerkships do not automatically provide an ideal learning environment for medical students. PMID- 10792688 TI - Comparison of anonymous student ballots with student debriefing for faculty evaluations. AB - INTRODUCTION: Yearly evaluation of academic faculty teaching is required by institutions for advancement purposes and continued employment. The method in which these evaluations are collected may influence the outcome of that evaluation. We compared the results of three different data collection methods of faculty ratings. METHODS: Diagnostic radiology residents evaluated four behaviour categories of faculty in three different ways during the 1995-96 academic year. The individual anonymous ballot was compared to two student debriefing techniques. RESULTS: Ratings in individual categories and rankings of several of the faculty changed considerably depending upon the data gathering method. Individual anonymous ballots produced a higher average rating in all four categories evaluated. The average ratings were lowest in the closed meeting group. DISCUSSION: The method in which evaluation of faculty are collected influences both the numerical value of the rating as well as the ranking of the teachers within the group. Evaluation outcomes are highly dependent upon the method of data collection. PMID- 10792689 TI - A process evaluation of medical ethics education in the first year of a new medical curriculum. AB - BACKGROUND: Despite the recent increase in activity in the field of medical ethics education, few evaluative studies have been carried out. Most studies have taken place in North America, in curricula where teaching is discipline-based, and have concentrated on outcome rather than on the curricular processes adopted. AIM: To evaluate the process of medical ethics education in the first year of a new learner-centred, problem-based, integrated medical curriculum. METHOD: A qualitative, multi-method approach was adopted using open questionnaires, focus groups and tutor evaluation rating scales. The study involved all 238 students in the first year of the new medical curriculum, and the 30 clinical tutors who facilitated ethics learning. A stratified sampling technique was used to choose focus group participants. RESULTS: Small group teaching proved highly acceptable to both students and tutors. Tutors' teaching skills were central to its effectiveness. Tutors played an important role in promoting students' appreciation of the relevance of medical ethics to clinical practice, and in establishing a climate where constructive criticism of colleagues' actions is acceptable. Course integration, including the provision for students of clinical experiences on which to reflect, was an important aid to learning. Students and tutors were noted to be driving the ethics curriculum towards having a contextual rather than theoretical base. CONCLUSION: This evaluation identified those aspects of the medical ethics course which contributed to its effectiveness and those which detracted from it. This information will be used to inform future development. PMID- 10792690 TI - An internet-based learning portfolio in resident education: the KOALA multicentre programme. AB - CONTEXT AND OBJECTIVES: To describe the Computerized Obstetrics and Gynecology Automated Learning Anaalysis (KOALAtrade mark), a multicentre, Internet-based learning portfolio and to determine its effects on residents' perception of their self-directed learning abilities. METHODS: The KOALA programme allows residents to record their obstetrical, surgical, ultrasound, and ambulatory patient encounters and to document critical incidents of learning or elements of surprise that arose during these encounters. By prompting the student to reflect on these learning experiences, KOALA encourages residents to articulate questions which can be directly pursued through hypertext links to evidence-based literature. Four Canadian residency training programmes participated in the pilot project, from February to May 1997, using a dynamic relational database with a central server. All participants completed the Self-directed Learning Readiness Scale and a learning habits questionnaire. The impact of the KOALA programme on residents' perception of their self-directed learning abilities was measured by comparing KOALA-naive schools (schools 2, 3, and 4) with school 1 (exposed to the KOALA prototype for 1 year). Ordered variables were compared using the Mann-Whitney U test and continuous variables with the Student t test (statistical significance P < 0. 05). RESULTS: During the study period, 7049 patient and 1460 critical incidents of learning were recorded by 41 residents in the four participating universities. Residents at the exposed school (school 1) had a significantly higher perception of their self-directed learning (P < 0.05) and believed their future learning was less likely to be from continuing medical education (P < 0.028), textbooks (P < 0.04), and didactic lectures (P < 0.011) and would be derived from a learning portfolio with online resources. CONCLUSION: This Internet-based, multi-user, multicentre learning portfolio has a significant effect on residents' perception of their self-directed learning abilities. PMID- 10792691 TI - Support and faculty mentoring programmes for medical students in Germany, Switzerland and Austria. AB - OBJECTIVES: To study what is being done at German-speaking universities regarding the counselling and tutoring of students, we carried out a survey among the deans of medical faculties in Germany, Austria and Switzerland. Our main concern was if any such projects were already available to the medical student or whether efforts to this purpose were under way. DESIGN: We focused in particular on faculty mentoring programmes, a continuous tutoring by designated members of the faculty on a person-to-person basis. SETTING: German, Austrian and Swiss medical faculties. SUBJECTS: Medical faculty deans. RESULTS: The return rate was 80%. While general student counselling is, if required, available at nearly all of the faculties, faculty mentoring programmes are offered by only 36.1% of the medical schools, and individualized career counselling by 30.6%. CONCLUSIONS: Compared to other countries, such as the United Kingdom or the United States, counselling and tutoring programmes, e.g. career planning or faculty mentoring, are not generally available to the German medical student. Regional differences are evident, which can be attributed to differences in the universities' legal and financial situation. The medical faculties at German-speaking universities should make it their priority to offer these services to the student on a permanent basis. PMID- 10792692 TI - Exploring the perceived effect of an undergraduate multiprofessional educational intervention. AB - CONTEXT: Improved teamwork and greater collaboration between professions are important factors in effective health care. These goals may be achieved by including interprofessional learning in the undergraduate medical curriculum. The Faculty of Medicine at the University of Liverpool organized a pilot two-day multiprofessional course involving all the health care related disciplines. OBJECTIVE: The present study examined the perceived effect of the multiprofessional course on the work practice of these newly qualified health care professionals. METHOD: The views of former students who took part in the pilot course were collected using a semi-structured interview schedule and analysed using a qualitative data analysis software package QSR NU*DIST. RESULTS: Two main themes emerged. These centred around role knowledge and interprofessional attitudes. Data indicated that participants perceived the course to have increased their knowledge of the other professions and that this effect had persisted. Reported benefits to their working practice included facilitating appropriate referrals, increasing professional empathy and awareness of other professionals' skills, raising confidence and heightening awareness of the holistic nature of patient treatment. Participants reported forming negative attitudes towards other professions during their undergraduate education. They believed these had been partly encouraged by course tutors. The pilot course was perceived to have had had little effect on these attitudes. Changes occurred once the newly qualified professionals started work. CONCLUSIONS: The results support the idea that interprofessional educational interventions must be tailored to specific learning goals to be implemented successfully, and that interprofessional education should be prolonged and widespread to have a real impact. PMID- 10792693 TI - Early effects of a new problem-based clinically oriented curriculum on students' perceptions of teaching. AB - OBJECTIVES: To compare the course experiences of medical students in a new problem-based (PBL) undergraduate medical course with those of their peers in a conventional curriculum. DESIGN: Whole class questionnaire survey using a pre validated research instrument. SETTING: University of Liverpool, UK. SUBJECTS: First and second year medical students RESULTS: New curriculum students were more satisfied with their course when compared to their conventional course peers. Problem solving, team working and motivation scores were significantly higher amongst new course (PBL) students. New course students were more anxious about clarity of objectives and standard of work required. CONCLUSIONS: Early evidence suggests that curriculum reform from conventional teaching to a small group problem analysis programme results in improvement in student satisfaction with teaching and the development of appropriate learning skills. PMID- 10792694 TI - Undergraduate medical students' views on the value of dissecting. PMID- 10792695 TI - Patterns of genetic variation in native grape phylloxera on two sympatric host species. AB - Random amplified polymorphic DNA (RAPD) markers were used to examine population genetic structure in populations of native grape phylloxera. This research asked: (i) do RAPD markers distinguish two groups corresponding to the two host plant species; and (ii) do RAPD markers distinguish groups according to spatial location, independent of host plant association? Forty-nine phylloxera clones were collected from five pairs of adjacent individuals of two sympatric grape species in five sites along a 145 km transect in Missouri, USA. A high level of polymorphism was observed, with some evidence for structuring between host plant species and no evidence for spatial structuring. An analysis of molecular variance (AMOVA) found that 6.52% of the variance in RAPD banding patterns was attributable to host species and 7.96% of the variance was attributable to spatial location. A cluster analysis did not result in two groups corresponding to the two hosts, or to five groups corresponding to the geographical sites sampled. A Mantel test showed a low correlation between genetic similarity and spatial location. Two of the 93 RAPD markers were nonrandomly associated between the hosts. It is suggested that there may be a small host-mediated effect on genetic variation but stochastic dispersal and a highly heterogeneous environment may be the primary influences on the observed polymorphism. PMID- 10792696 TI - Parentage analysis with incomplete sampling of candidate parents and offspring. AB - Many breeding systems include 'multiple mating' in which males or females mate with multiple partners. We identify two forms of multiple mating: 'single-sex', where the next-generation individuals (NGIs) are the product of multiple mating by one sex; and 'two-sex', where the NGIs are the product of multiple mating by both sexes. For both mating systems we develop models that estimate the proportion of NGIs that is fathered (paternity) or mothered (maternity) by the putative parents. The models only require genetic data from the parent or parents in question and the sample of NGIs, as well as an estimate of population allele frequencies. The models provide unbiased estimates, can accommodate loci with many alleles and are robust to violations of their assumptions. They allow researchers to address intractable problems such as the parentage of seeds found on the ground, juvenile fish in a stream, and nestlings in a communal breeding bird. We demonstrate the models using genetic data from a nest of the bluegill sunfish Lepomis macrochirus, where the NGIs may be from multiple females that have spawned with multiple males from different life histories (cuckolder and parental). PMID- 10792697 TI - Statistical confidence in parentage analysis with incomplete sampling: how many loci and offspring are needed? AB - We have recently presented models to estimate parentage in breeding systems with multiple mating and incomplete sampling of the candidate parents. Here we provide formulas to calculate the statistical confidence and the optimal trade-off between the number of loci and offspring. These calculations allow an understanding of the statistical significance of the parentage estimates as well as the appropriate sampling regime required to obtain a desired level of confidence. We show that the trade-off generally depends on the parentage of the putative parents. When parentage is low, sampling effort should concentrate on increasing the number of loci. Otherwise, there are similar benefits from increasing the number of loci or offspring. We demonstrate these methods using genetic data from a nest of the bluegill sunfish (Lepomis macrochirus). PMID- 10792699 TI - Nested cladistic analysis, phylogeography and speciation in the Timarcha goettingensis complex (Coleoptera, chrysomelidae). AB - The Timarcha goettingensis complex is a monophyletic assemblage of closely related leaf beetles (Chrysomelidae), distributed from the north half of the Iberian Peninsula to Central Europe. Oligophagy, mountainous habitat and apterism are factors which are assumed to promote speciation in these beetles. We have used cytochrome oxidase subunit II mitochondrial DNA genealogies obtained from 31 sampling localities and a nested geographical distance analysis to assess the population structure and demographic factors explaining the geographical distributions of the mtDNA haplotypes in the T. goettingensis complex. The results show that there is a significant association between genetic structuring and geography. Inferences about the historical population processes in the species complex are discussed, being in general in accordance with contiguous range expansions and past fragmentations. The use of the cohesion species concept approach suggests the existence of several systematic ranks among the different T. goettingensis populations, which is in part supported by ecological traits such as trophic selection and altitudinal distribution. PMID- 10792698 TI - Origin and evolution of invasive naturalized material of Rhododendron ponticum L. in the British isles. AB - Information concerning the area of origin, genetic diversity and possible acquisition of germplasm through hybridization is fundamental to understanding the evolution, ecology and possible control measures for an introduced invasive plant species. Rhododendron ponticum is extensively naturalized in the British Isles, but it is not known whether native material in Turkey, Spain or Portugal gave rise to the naturalized material, or to what extent introgression has affected this material. Chloroplast (cp) and nuclear ribosomal DNA (rDNA) restriction fragment length polymorphisms (RFLPs) were sought which could distinguish between native material of R. ponticum, and between 15 other Rhododendron species including R. ponticum's closest relatives. Thereafter, a total of 260 naturalized accessions of R. ponticum from throughout the British Isles was examined with respect to informative polymorphisms. It was found that 89% of these accessions possessed a cpDNA haplotype that occurred in native material of R. ponticum derived almost entirely from Spain, while 10% of accessions had a haplotype unique to Portuguese material. These results therefore indicated an Iberian origin for British material. rDNA or cpDNA evidence of introgression from R. catawbiense was found in 27 British accessions of R. ponticum, and such accessions were significantly more abundant in Britain's coldest region, eastern Scotland, than elsewhere. This could indicate that introgression from R. catawbiense confers improved cold tolerance. Introgression from R. maximum and an unidentified species was also detected. PMID- 10792700 TI - An analysis of genetic variation in natural populations of Sticherus flabellatus [R. Br. (St John)] using amplified fragment length polymorphism (AFLP) markers. AB - Amplified fragment length polymorphisms (AFLPs) were used to characterize the genetic diversity within and among natural populations of Sticherus flabellatus. Eight populations within the Sydney region of New South Wales, Australia were surveyed using 11 primer combinations. A total of 1108 reproducible bands were detected of which 469 (42%) were polymorphic. FST estimates averaged over all polymorphic loci indicated that significant genomic differentiation occurs among populations (average = 0.783). Genetic diversity within populations was assessed according to average heterozygosity (H) and percentage polymorphic loci (P) per population. Within-population diversity ranged from H = 0.12 and P = 33.69 to H = 0.04 and P = 15.99. Analysis of genetic similarity among populations suggested that the eight populations studied fall into two groups of four populations, based on population size and the condition of the habitat. Phenetic analysis (AMOVA) indicated that genetic variation is greater among populations (74.34%) than within populations (25.66%). These findings suggest that the breeding system of S. flabellatus is predominantly inbreeding, with genetic diversity maintained by occasional outcrossing in larger populations. The results presented in this study could provide evidence to support the proposal to protect natural stands of S. flabellatus, which has implications for the Australian horticulture industry. PMID- 10792701 TI - Microsatellite and mitochondrial DNA polymorphism reveals life-history dependent interbreeding between hatchery and wild brown trout (Salmo trutta L.). AB - The effects of stocking hatchery trout into wild populations were studied in a Danish river, using microsatellite and mitochondrial DNA (mtDNA) markers. Baseline samples were taken from hatchery trout and wild trout assumed to be unaffected by previous stocking. Also, samples were taken from resident and sea trout from a stocked section of the river. Genetic differentiation between the hatchery strain and the local wild population was modest (microsatellite FST = 0.06). Using assignment tests, more than 90% of individuals from the baseline samples were classified correctly. Assignment tests involving samples from the stocked river section suggested that the contribution by hatchery trout was low among sea trout (< 7%), but high (46%) among resident trout. Hybrid index analysis and a high percentage of mtDNA haplotypes specific to indigenous trout observed among resident trout that were assigned to the hatchery strain suggested that interbreeding took place between hatchery and wild trout. The latter result also indicated that male hatchery trout contributed more to interbreeding than females. We suggest that stronger selection acts against stocked hatchery trout that become anadromous compared to hatchery trout that become resident. As most resident trout are males this could also explain why gene flow from hatchery to wild trout appeared to be male biased. The results show that even despite modest differentiation at neutral loci domesticated trout may still perform worse than local populations and it is important to be aware of differential survival and reproductive success both between life-history types and between sexes. PMID- 10792702 TI - Molecular phylogeography and cryptic speciation in the mosses, Mielichhoferia elongata and M. mielichhoferiana (Bryaceae). AB - Nucleotide sequence variation in the ITS1-5.8S-ITS2 region of nuclear ribosomal DNA (nrDNA) from 70 populations of Mielichhoferia elongata and M. mielichhoferiana, plus two outgroup species, was analysed using maximum parsimony and maximum likelihood methods. High levels of nucleotide substitution and numerous insertion-deletion events were detected within and between the two species. M. elongata is monophyletic with regard to nrDNA variation, but M. mielichhoferiana is paraphyletic. (M. elongata is nested within it.) A clade within M. mielichhoferiana provides evidence of vicariance, with North American and Scandinavian sister groups of populations. Two major clades are resolved in M. elongata by sequence data that are completely congruent with previous isozyme work. One clade includes populations from both North America and Europe whereas the other is strictly North American. These two clades, resolved by multiple independent loci, clearly represent cryptic species within the morphologically uniform M. elongata. Certain geographical areas, most notably southwestern Colorado in Ouray and San Juan Counties, harbour diverse populations of M. elongata with distinct phylogenetic and phylogeographical histories. Morphologically indistinguishable but phylogenetically distant populations were detected a few metres apart at one site. In contrast, all populations collected over hundreds of kilometres in California belong to a single clade. Arctic North American populations belong to a clade that includes disjunct populations in Alaska, northern Ellesmere Island, and the northeastern USA, but not subarctic Swedish populations, which are more closely related to plants from the Rocky Mountains. Morphological uniformity belies complex infraspecific phylogenetic patterns within M. elongata and M. mielichhoferiana. PMID- 10792703 TI - Genetic distinctness of isolated populations of an endangered marsupial, the mountain pygmy-possum, Burramys parvus. AB - The mountain pygmy-possum, Burramys parvus, exists in isolated and fragmented populations in the Australian alps. To examine the degree of interpopulation divergence, mitochondrial cytochrome b and NADH dehydrogenase subunit 2 (NADH2) sequences were obtained from samples representing all populations of B. parvus. Three divergent mitochondrial DNA (mtDNA) lineages were identified which exhibited strong phylogeographical structure. This indicates the presence of three maternal clades corresponding to populations in the northern, central and southern Australian alps. Molecular clock estimates suggest that the mtDNA lineages diverged from one another 420-680 thousand years ago. On this basis it is argued that B. parvus populations have probably been isolated since the mid Pleistocene, and that management should focus on maintaining viable B. parvus populations in each of the three regional localities. PMID- 10792704 TI - Ecological determinants and temporal stability of the within-river population structure in Atlantic salmon (Salmo salar L.). AB - A gene diversity analysis was performed using microsatellite loci in order to (i) describe the extent and pattern of population structure in Atlantic salmon (Salmo salar L.) within a river system; (ii) establish the importance of quantifying the signal:noise ratio in accurately estimating population structure; and (iii) assess the potential usefulness of two evolutionary models in explaining within river population structure from the ecological and habitat characteristics of Atlantic salmon. We found weak, yet highly significant microscale spatial patterning after accounting for variance among temporal replicates within sites. Lower genetic distances were observed among temporal samples at four sampling sites whereas no evidence for temporal stability was observed at the other three locations. The component of genetic variance attributable to either temporal instability and/or random sampling errors was almost three times more important than the pure spatial component. This indicates that not considering signal:noise ratio may lead to an important overestimation of genetic substructuring in situations of weak genetic differentiation. This study also illustrates the usefulness of the member-vagrant hypothesis to generate a priori predictions regarding the number of subpopulations that should compose a species, given its life-history characteristics and habitat structure. On the other hand, a metapopulation model appears better suited to explain the extent of genetic divergence among subpopulations, as well as its temporal persistence, given the reality of habitat patchiness and environment instability. We thus conclude that the combined use of both models may offer a promising avenue for studies aiming to understand the dynamics of genetic structure of species found in unstable environments. PMID- 10792705 TI - Microsatellite loci isolated from Odontesthes argentinensis and the O. perugiae species group and their use in other South American silverside fish. PMID- 10792707 TI - Characterization of microsatellite loci developed for the wattled curassow, Crax globulosa. PMID- 10792706 TI - Microsatellite characterization in the rainbow wrasse Coris julis (Pisces: labridae). PMID- 10792708 TI - Microsatellites in rockfish Sebastes thompsoni (Scorpaenidae). PMID- 10792709 TI - Characterization of microsatellite loci in the endangered freshwater crayfish Austropotamobius pallipes (Astacidae) and their potential use in other decapods. PMID- 10792710 TI - Microsatellite loci from common and thick-billed murres, Uria aalge and U. lomvia. PMID- 10792711 TI - Development of polymorphic microsatellite markers in a tropical tree species, Melaleuca cajuputi (Myrtaceae). PMID- 10792712 TI - Microsatellite markers for investigating population structure in octopus vulgaris (Mollusca: cephalopoda). PMID- 10792713 TI - Characterization of microsatellite loci in bannertailed and giant kangaroo rats, Dipodomys spectabilis and Dipodomys ingens. PMID- 10792714 TI - Ralstonia solanacearum produces hrp-dependent pili that are required for PopA secretion but not for attachment of bacteria to plant cells. AB - As in many other Gram-negative plant pathogenic bacteria, the Ralstonia solanacearum hrp genes are involved in the production of a type III secretion apparatus that allows the translocation of PopA protein to the external medium. Here, we show that hrp genes are also involved in the biogenesis of pili that are mainly composed of the HrpY protein. These pili are produced at one pole of the bacterium and are also released into the external medium where they can form very long straight bundles. An hrpY mutant is defective in pilus production, impaired in interactions with plants and in secretion of the PopA protein but not in attachment to plant cells. PMID- 10792715 TI - Two novel proteins, PopB, which has functional nuclear localization signals, and PopC, which has a large leucine-rich repeat domain, are secreted through the hrp secretion apparatus of Ralstonia solanacearum. AB - The Ralstonia solanacearum hrp gene cluster codes for components of a type III secretion pathway necessary for the secretion of PopA1, a hypersensitive response like elicitor protein. In the present study, we show that several other Hrp secreted proteins can be detected by growing wild-type bacteria in minimal medium in the presence of Congo red. Two of these proteins, PopB and PopC, are encoded by genes located downstream of popA and constitute an operon with popA. popABC mutants retain the wild-type ability to cause disease in hosts and to elicit the hypersensitive response on non-hosts. Expression of the popABC operon is controlled by the hrpB regulatory gene and is induced upon co-culture with Arabidopsis cell suspensions. This plant cell-specific induction depends on PrhA, a putative receptor for plant specific signal(s). The transcription of the popABC operon is not modified by the addition of Congo red to the growth medium and the intracellular pools of PopB and PopC are very similar in the absence or presence of Congo red. Preliminary data suggest that Congo red stabilizes secreted proteins in the extracellular medium. PopB is a 173-amino-acid-basic protein that contains a functional bipartite nuclear localization signal. PopC is a 1024-amino acid protein that carries 22 tandem leucine-rich repeats (LRR). The LRR domain of this protein forms a consensus that perfectly matches the predicted eukaryotic cytoplasmic LRR consensus. We propose that PopB and PopC may be translocated into plant cells via the Hrp pathway. PMID- 10792716 TI - Intrinsic instability of the essential cell division protein FtsL of Bacillus subtilis and a role for DivIB protein in FtsL turnover. AB - Cell division in most eubacteria is driven by an assembly of about eight conserved division proteins. These proteins form a ring structure that constricts in parallel with the formation of the division septum. Here, we show that one of the division proteins, FtsL, is highly unstable. We also show that the protein is targeted to the ring structure and that targeting occurs in concert with the recruitment of several other membrane-associated division proteins. FtsL stability is further reduced in the absence of DivIB protein (probably homologous to E. coli FtsQ) at high temperature, suggesting that DivIB is involved in the control of FtsL turnover. The reduced stability of FtsL may explain the temperature dependence of divIB mutants, because their phenotype can be suppressed by overexpression of FtsL. The results provide new insights into the roles of the FtsL and DivIB proteins in bacterial cell division. PMID- 10792717 TI - Aspergillus SteA (sterile12-like) is a homeodomain-C2/H2-Zn+2 finger transcription factor required for sexual reproduction. AB - Saccharomyces cerevisiae Ste12p plays a key role in coupling signal transduction through MAP kinase modules to cell-specific or morphogenesis-specific gene expression required for mating and pseudohyphal (PH)/filamentous growth (FG). Ste12p homologues in the pathogenic yeasts Candida albicans and Filobasidiela neoformans apparently play similar roles during dimorphic transitions. Here we report the isolation and characterization of the first Ste12 protein from a true filamentous fungus. Aspergillus nidulans steA encodes a protein with a homeodomain 63-75% identical to those of other Ste12 proteins, with greatest similarity to FnSte12alphap. SteAp and Ste12alphap lack the pheromone induction domain found in budding yeast Ste12p, but have C-terminal C2/H2-Zn+2 finger domains not present in the other Ste12 proteins. A DeltasteA strain is sterile and differentiates neither ascogenous tissue nor fruiting bodies (cleistothecia). However, the development of sexual cycle-specific Hulle cells is unaffected. Filamentous growth, conidiation and the differentiation of PH-like asexual reproductive cells (metulae and phialides) are normal in the deletion strain. Northern analysis of key regulators of the asexual and sexual reproductive cycles support the observation that although SteAp function is restricted to the sexual cycle, cross regulation between the two developmental pathways exists. Our results further suggest that while several classes of related proteins control similar morphogenetic events in A. nidulans and the dimorphic yeasts, significant differences must exist in the regulatory circuitry. PMID- 10792718 TI - Identification of an AfsA homologue (BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis, virginiamycin biosynthesis and virginiamycin M1 resistance. AB - Virginiae butanolide (VB)-BarA of Streptomyces virginiae is one of the newly discovered pairs of a gamma-butyrolactone autoregulator and the corresponding receptor protein of the Streptomyces species, and has been shown to regulate the production of antibiotic virginiamycin (VM) in S. virginiae. A divergently transcribed barX gene is situated 259 bp upstream of the barA gene, and the BarX protein has been shown to be highly homologous (39.8% identity, 74. 6% similarity) to S. griseus AfsA. Although AfsA is thought to be a biosynthetic enzyme for A-factor, another member of the family of gamma-butyrolactone autoregulators, the in vivo function of S. virginiae BarX was investigated in this study by phenotypic and transcriptional comparison between wild-type S. virginiae and a barX deletion mutant. With the same growth rate as wild-type S. virginiae on both solid and liquid media, the barX mutant showed no apparent changes in its morphological behaviour, indicating that barX does not participate in morphological control in S. virginiae. However, the barX mutant became more sensitive to virginiamycin M1 than did the wild-type strain (minimum inhibitory concentration, 50 microgram ml-1 compared with > 200 microgram ml-1) and exhibited reduced VB and VM production. The VM production was not restored by exogenous addition of VB, suggesting that BarX per se is not a biosynthetic enzyme of VBs but a pleiotropic regulatory protein controlling VB biosynthesis. DNA sequencing of a 5.6 kbp downstream region of barX revealed the presence of five open reading frames (ORFs): barZ, encoding a BarB-like regulatory protein; orf2, encoding a Streptomyces coelicolor RedD-like pathway specific regulator; varM, encoding a homologue of ATP-dependent transporters for macrolide antibiotics; orf4, encoding a homologue of beta-ketoacyl ACP/CoA reductase; and orf5, encoding a homologue of dNDP-glucose dehydratase. Reverse transcription polymerase chain reaction (RT-PCR) analyses of the downstream five genes together with those of the three upstream genes (barA, barB, encoding a regulatory protein; and varS, encoding a virginiamycin S specific transporter) revealed that, in the barX mutant, the transcriptions of barZ, orf2, varM and orf5 were completely repressed and those of barB and varS were derepressed. Because free BarA (BarA in the absence of VB) in wild-type S. virginiae represses the transcription of bicistronic barB-varS operon through binding to a specific DNA sequence (BarA-responsive element, BARE) overlapping the barB transcriptional start site, the derepression of barB-varS transcription in the barX mutant suggested that the in vivo function of BarA was impaired by the lack of BarX protein. Gel-shift assays revealed that BarA easily lost its DNA-binding activity in the absence of BarX but that the defect was restored by the presence of recombinant BarX as a fusion with maltose-binding protein (MBP-BarX), whereas MBP BarX itself showed no DNA-binding activity, indicating that BarX is likely to be a co-repressor of BarA, enforcing the DNA-binding activity of BarA through protein-protein interactions. PMID- 10792719 TI - Membrane-to-cytosol redistribution of ECF sigma factor AlgU and conversion to mucoidy in Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - The conversion to mucoid phenotype in Pseudomonas aeruginosa during chronic infections in cystic fibrosis (CF) is due to mutations in the algU mucABCD gene cluster. This cluster encodes an extreme stress response system conserved in Gram negative bacteria. The system includes an ECF sigma factor, AlgU (sigmaE), an inner membrane protein, MucA, which inhibits AlgU activity, and MucB, a periplasmic protein that negatively controls AlgU. In this work, we investigated whether and how these factor interact to transduce signals between different cellular compartments. The mutation mucADeltaG440, which renders a large fraction of P. aeruginosa CF isolates mucoid, did not abrogate AlgU-MucA interactions, although it eliminated MucA-MucB interactions in the yeast two-hybrid system. The mucADeltaG440 truncation of the periplasmic C-terminal tail of MucA destabilized the molecule resulting in low or undetectable steady-state levels in P. aeruginosa. Somewhat reduced levels of MucA were also seen in cells with inactivated mucB or with the mucACF53 allele carrying the missense P184S mutation, which mildly affected interactions with MucB. The events downstream from MucA destabilization were also investigated. AlgU was found to associate with inner membranes in mucA+ cells. In mutants destabilizing MucA, a limited redistribution of AlgU from the membrane to the cytosol was observed. The redistribution was spontaneous in mucADeltaG440 cells, while in mucB and mucACF53 mutants it required additional signals. Despite a large reduction in MucA levels in mucADeltaG440 cells, only a small fraction of AlgU was redistributed to the cytosol and a significant portion of this sigma factor remained membrane bound and behaved as a peripheral inner membrane protein. The fraction of AlgU that depended on MucA for association with the membrane also brought RNA polymerase into this compartment. These results are consistent with a model in which MucB MucA-AlgU-RNA polymerase interactions at the membrane allow transduction of potentially lethal stress signals with both rapid reaction times of the preassembled complexes and efficient resupply at the membrane from the prebound components. PMID- 10792720 TI - Differential RNA elongation controls the variant surface glycoprotein gene expression sites of Trypanosoma brucei. AB - The protozoan parasite Trypanosoma brucei develops antigenic variation to escape the immune response of its host. To this end, the trypanosome genome contains multiple telomeric expression sites competent for transcription of variant surface glycoprotein genes, but as a rule only a single antigen is expressed at any time. We used reverse transcription-PCR (RT-PCR) to analyse transcription of different segments of the expression sites in different variant clones of two independent strains of T. brucei. The results indicated that RNA polymerase is installed and active at the beginning of many, if not all, expression sites simultaneously, but that a progressive arrest of RNA elongation occurs in all but one site. This defect is linked to inefficient RNA processing and RNA release from the nucleus. Therefore, functional transcription in the active site appears to depend on the selective recruitment of a RNA elongation/processing machinery. PMID- 10792721 TI - Dual regulation of mucoidy in Pseudomonas aeruginosa and sigma factor antagonism. AB - The conversion to mucoid, exopolysaccharide alginate-overproducing phenotype in Pseudomonas aeruginosa during chronic respiratory infections in cystic fibrosis patients occurs via mutations that activate the alternative sigma factor AlgU (sigmaE). In this study, we demonstrate that conversion to mucoidy can be caused via a second, algU-independent pathway, in which alginate production and transcription of the critical algD promoter depend on another alternative sigma factor, RpoN (sigma54). The algD promoters dependent on sigma54 and sigmaE showed a complete overlap resulting in identical mRNA 5' ends. The two pathways were not independent, as sigma54 also repressed sigmaE-dependent transcription of algD both in vitro and in vivo. The negative regulatory effect of sigma54 on sigmaE dependent algD expression was based on sigma54 binding to the algD promoter and its interference with sigmaE-dependent transcription. This phenomenon, referred to here as sigma factor antagonism, reflects the unique properties of sigma54, which lacks an intrinsic ability to form open transcription initiation complexes. We propose that this peculiar feature of sigma54 has evolved in part to allow its recruitment as a repressor of certain promoter subsets. The repression of algD by sigma54 also depends on environmental conditions, supporting the notion that sigma factor antagonism plays a physiological role in controlling alginate production in P. aeruginosa during adaptation to different ecological sites (e.g. biofilm development, stress and other growth conditions) and unique environments in the chronically infected host. PMID- 10792722 TI - RAS1 regulates filamentation, mating and growth at high temperature of Cryptococcus neoformans. AB - Cryptococcus neoformans is a basidiomycete yeast and opportunistic human pathogen of increasing clinical importance due to the increasing population of immunocompromised patients. To further investigate signal transduction cascades regulating fungal pathogenesis, we have identified the gene encoding a RAS homologue in this organism. The RAS1 gene was disrupted by transformation and homologous recombination. The resulting ras1 mutant strain was viable, but failed to grow at 37 degrees C, and exhibited significant defects in mating and agar adherence. The ras1 mutant strain was also avirulent in an animal model of cryptococcal meningitis. Reintroduction of the wild-type RAS1 gene complemented these ras1 mutant phenotypes and restored virulence in animals. A dominantly active RAS1 mutant allele, RAS1Q67L, induced a differentiation phenotype known as haploid fruiting, which involves filamentation, agar invasion and sporulation in response to nitrogen deprivation. The ras1 mutant mating defect was suppressed by overexpression of MAP kinase signalling elements and partially suppressed by exogenous cAMP. Additionally, cAMP also suppressed the agar adherence defect of the ras1 mutant. However, the ability of the ras1 mutant strain to grow at elevated temperature was not restored by cAMP or MAP kinase overexpression. Our findings support a model in which RAS1 signals in C. neoformans through cAMP dependent, MAP kinase, and RAS-specific signalling cascades to regulate mating and filamentation, as well as growth at high temperature which is necessary for maintenance of infection. PMID- 10792723 TI - Extracellular targeting of choline-binding proteins in Streptococcus pneumoniae by a zinc metalloprotease. AB - A genetic-based search for surface proteins of Streptococcus pneumoniae involved in adhesion identified a putative zinc metalloprotease (ZmpB). ZmpB shared high amino acid sequence similarities with IgA1 proteases of Gram-positive bacteria, but ZmpB had neither IgA1 nor IgA2 protease activity. Analysis of a family of surface-expressed proteins, the choline-binding proteins (Cbp's), in a zmpB deficient mutant demonstrated a global loss of surface expression of CbpA, CbpE, CbpF and CbpJ. CbpA was detected within the cytoplasm. The zmpB-deficient mutant also failed to lyse with penicillin, a sign of lack of function of the Cbp LytA. Immunodetection studies revealed that the autolysin (LytA), normally located on the cell wall, was trapped in the cytoplasm colocalized with DNA and the transformation protein CinA. Trafficking of CinA and RecA to the cell membrane during genetic competence was also not observed in the zmpB-deficient mutant. These results suggest a protease dependent regulatory mechanism governing the translocation of CinA and the Cbp's LytA and CbpA of S. pneumoniae. PMID- 10792724 TI - Sxa2 is a serine carboxypeptidase that degrades extracellular P-factor in the fission yeast Schizosaccharomyces pombe. AB - Stimulating the fission yeast Schizosaccharomyces pombe with mating pheromones brings about responses that lead to cell conjugation. Persistent stimulation does not, however, induce a continuous response as the cells become desensitized to the presence of the pheromone. One mechanism that contributes to desensitization in M-cells is the release of a carboxypeptidase that inactivates the extracellular P-factor pheromone. Production of the carboxypeptidase requires a functional sxa2 gene. In this study, we report the first molecular characterization of the Sxa2 protein and provide direct evidence that it is the carboxypeptidase that degrades P-factor. Sxa2 is synthesized as a precursor that undergoes an internal cleavage event catalysed by a protease with specificity for basic residues. This generates a series of catalytically active N-terminal fragments and an inactive C-terminal fragment. Cleavage is essential for activation of the carboxypeptidase and, although the C-terminal fragment is inactive, it is required for the N-terminal fragment to attain activity. PMID- 10792725 TI - A defined system for hybrid macrolide biosynthesis in Saccharopolyspora erythraea. AB - The biological activity of polyketide antibiotics is often strongly dependent on the presence and type of deoxysugar residues attached to the aglycone core. A system is described here, based on the erythromycin-producing strain of Saccharopolyspora erythraea, for detection of hybrid glycoside formation, and this system has been used to demonstrate that an amino sugar characteristic of 14 membered macrolides (D-desosamine) can be efficiently attached to a 16-membered aglycone substrate. First, the S. erythraea mutant strain DM was created by deletion of both eryBV and eryCIII genes encoding the respective ery glycosyltransferase genes. The glycosyltransferase OleG2 from Streptomyces antibioticus, which transfers L-oleandrose, has recently been shown to transfer rhamnose to the oxygen at C-3 of erythronolide B and 6-deoxyerythronolide B. In full accordance with this finding, when oleG2 was expressed in S. erythraea DM, 3 O-rhamnosyl-erythronolide B and 3-O-rhamnosyl-6-deoxyerythronolide B were produced. Having thus validated the expression system, endogenous aglycone production was prevented by deletion of the polyketide synthase (eryA) genes from S. erythraea DM, creating the triple mutant SGT2. To examine the ability of the mycaminosyltransferase TylM2 from Streptomyces fradiae to utilise a different amino sugar, tylM2 was integrated into S. erythraea SGT2, and the resulting strain was fed with the 16-membered aglycone tylactone, the normal TylM2 substrate. A new hybrid glycoside was isolated in good yield and characterized as 5-O-desosaminyl-tylactone, indicating that TylM2 may be a useful glycosyltransferase for combinatorial biosynthesis. 5-O-glucosyl-tylactone was also obtained, showing that endogenous activated sugars and glycosyltransferases compete for aglycone in these cells. PMID- 10792726 TI - Hyperactive forms of the Pdr1p transcription factor fail to respond to positive regulation by the hsp70 protein Pdr13p. AB - Multidrug resistance in Saccharomyces cerevisiae is commonly associated with the overproduction of ATP-binding cassette transporter proteins such as Pdr5p or Yor1p. The Cys6-Zn(II)2 cluster-containing transcription factors Pdr1p and Pdr3p are key regulators of expression of these pleiotropic drug resistance (PDR) loci. Previous experiments have demonstrated that the Hsp70 protein encoded by the PDR13 gene is a positive regulator of Pdr1p function. We have examined the mechanism underlying the control of Pdr1p by Pdr13p. Expression of deletion, insertion and amino acid substitution mutant variants of Pdr1p suggest that the centre region of the transcription factor is the target for Pdr13p-mediated positive regulation. Immunological and fusion protein analyses demonstrate that Pdr13p is located in the cytoplasm, while Pdr1p is found in the nucleus. Biochemical fractionation experiments indicate that Pdr13p is associated with a high-molecular-weight complex and suggest the association of some fraction of Pdr13p with ribosomes. PMID- 10792727 TI - The G-protein FlhF has a role in polar flagellar placement and general stress response induction in Pseudomonas putida. AB - The flhF gene of Pseudomonas putida, which encodes a GTP-binding protein, is part of the flagellar-motility-chemotaxis operon. Its disruption leads to a random flagellar arrangement in the mutant (MK107) and loss of directional motility in contrast to the wild type, which has polar flagella. The return of a normal flhF allele restores polar flagella and normal motility to MK107; its overexpression triples the flagellar number but does not restore directional motility. As FlhF is homologous to the receptor protein of the signal recognition particle (SRP) pathway of membrane protein translocation, this pathway may have a role in polar flagellar placement in P. putida. MK107 is also compromised in the development of the starvation-induced general stress resistance (SGSR) and effective synthesis of several starvation and exponential phase proteins. While somewhat increased protein secretion in MK107 may contribute to its SGSR impairment, the altered protein synthesis pattern also appears to have a role. PMID- 10792728 TI - Specificity determinants for bacteriophage Hong Kong 022 integrase: analysis of mutants with relaxed core-binding specificities. AB - The integrase (Int) proteins encoded by bacteriophages HK022 and lambda catalyse similar site-specific integration and excision reactions between specific DNA regions known as attachment (att) sites. However, the Int proteins of HK022 and lambda are unable to catalyse recombination between non-cognate att sites. The att sites of both phages contain weak binding sites for Int, known as 'core-type' sites. Negatively acting nucleotide determinants associated with specific core sites (lambda B', HK022 B', HK022 C) are responsible for the barrier to non cognate recombination. In this study, we used challenge phages to demonstrate that the lambda and HK022 Ints cannot bind to core sites containing non-cognate specificity determinants in vivo. We isolated mutants of the HK022 Int, which bind the lambda B' core site. Two mutants, D99N and D99A, have changed a residue in the core-binding (CB) domain, which may be directly contacting the core site DNA. We suggest that binding to the lambda B' site was accomplished by removing the negatively charged aspartate residue, which normally participates in a conflicting interaction with the G4 nucleotide of the lambda B' site. We showed that, although our mutants retain the ability to recombine their cognate att sites, they are unable to recombine lambda att sites. PMID- 10792729 TI - Repair of double-strand breaks by incorporation of a molecule of homologous DNA. AB - An in vitro system based upon extracts of Escherichia coli infected with bacteriophage T7 was used to monitor repair of double-strand breaks in the T7 genome. The efficiency of double-strand break repair was markedly increased by DNA molecules ('donor' DNA) consisting of a 2.1 kb DNA fragment, generated by PCR, that had ends extending approximately 1 kb on either side of the break site. Repair proceeded with greater than 10% efficiency even when T7 DNA replication was inhibited. When the donor DNA molecules were labelled with 32P, repaired genomes incorporated label only near the site of the double-strand break. When repair was carried out with unlabelled donor DNA and [32P]-dCTP provided as precursor for DNA synthesis the small amount of incorporated label was distributed randomly throughout the entire T7 genome. Repair was performed using donor DNA that had adjacent BamHI and PstI sites. When the BamHI site was methylated and the PstI site was left unmethylated, the repaired genomes were sensitive to PstI but not to BamHI endonuclease, showing that the methyl groups at the BamHI recognition site had not been replaced by new DNA synthesis during repair of the double-strand break. These observations are most consistent with a model for double-strand break repair in which the break is widened to a small gap, which is subsequently repaired by physical incorporation of a patch of donor DNA into the gap. PMID- 10792730 TI - Inhibition of pyruvate-ferredoxin oxidoreductase gene expression in Giardia lamblia by a virus-mediated hammerhead ribozyme. AB - Giardia lamblia is a primitive eukaryotic microorganism that derives its metabolic energy primarily from anaerobic glycolysis. In trophozoites, pyruvate ferredoxin oxidoreductase (PFOR) converts pyruvate to acetyl-CoA with the transfer of a pair of electrons to ferredoxin, which can then reduce metronidazole and activate it into a potent antigiardiasis agent. It is unclear, however, whether this anaerobic disposal of electrons is essential for the energy metabolism in Giardia. In the present study, cDNAs encoding hammerhead ribozyme flanked with various lengths of antisense PFOR RNA were cloned into a viral vector pC631pac derived from the genome of giardiavirus (GLV). RNA transcripts of the plasmids showed high cleavage activities on PFOR mRNA in vitro. They were introduced into GLV-infected G. lamblia trophozoites by electroporation and stablized in the transfected cells via serial passages under puromycin selection. PFOR mRNA and enzyme activity in the transfected cells were decreased by 46-60% with the ribozyme PRzS flanked with 20 nt PFOR antisense RNA on each arm and by 69-80% with the ribozyme PRzL flanked with 600 and 1500 nt PFOR antisense RNA. PRzS without the inserted ribozyme or ribozyme flanked with alcohol dehydrogenase E antisense RNA showed no effect on PFOR mRNA and activity. The ribozyme transfected cells demonstrated significantly enhanced resistance to metronidazole and grew equally well under anaerobic and aerobic conditions. In contrast, the wild-type cells grew slightly better anaerobically than the transfectants but did not grow at all in aerobic conditions. Thus, the reduced PFOR expression enables Giardia to grow under molecular oxygen and the presence of PFOR enhances the anaerobic growth of Giardia with an increased susceptibility towards metronidazole. In addition, this study demonstrated for the first time the feasibility of using a viral RNA vector to express a ribozyme targeted at a specific mRNA in G. lamblia to reduce the expression of a specific gene. PMID- 10792731 TI - Effects of local transcription and H-NS on inversion of the fim switch of Escherichia coli. AB - The fim switch of Escherichia coli is responsible for phase-variable expression of type 1 fimbriae. Switching in the ON-to-OFF and OFF-to-ON directions is promoted by the FimB recombinase, while the FimE recombinase directs switching predominantly in the ON-to-OFF direction. The effects of local promoter activity and the H-NS nucleoid-associated protein on inversion of the switch were assessed. In contrast to FimB-mediated inversion, inversion of the switch by the FimE recombinase was unaffected by the H-NS status of the cell. Transcription towards the switch from within a translationally inactivated fimE gene was found to bias the switch strongly in the OFF direction, creating a FimE+-like phenotype in the absence of the FimE protein. This biasing was H-NS dependent and was also contingent on transcription from within the switch. These data show that local transcription and a nucleoid-associated protein both contribute to the modulation of a site-specific recombination event on the bacterial chromosome. PMID- 10792732 TI - Staphylococcal alpha-toxin: repair of a calcium-impermeable pore in the target cell membrane. AB - Staphylococcal alpha-toxin forms heptameric pores that render membranes permeable for monovalent cations. The pore is formed by an amphipathic beta-barrel encompassing amino acid residues 118-140 of each subunit of the oligomer. Human fibroblasts are susceptible to alpha-toxin but are able to repair the membrane lesions. Thereby, toxin oligomers remain embedded in the plasma membrane and exposed to the extracellular medium. In this study, we sought to detect structural changes occurring in the pore-forming sequence during lesion repair. Single cysteine substitution mutants were labelled with the environmentally sensitive fluorochrome acrylodan and, after mixing with wild-type toxin, incorporated into hybrid heptamers on fibroblast membranes. Formation of the lipid-inserted beta-barrel was accompanied by characteristic fluorescence emission shifts. After lesion repair, the environment of the residues at the outer surface of the beta-barrel remained unchanged, indicating continued contact with lipids. However, the labelled residues oriented towards the channel lumen underwent a green to blue shift in fluorescence, indicating reduced exposure to water. Pore closure proceeded in the presence of calmodulin inhibitors and of microtubule disruptors; however, it was prevented by cytochalasin D and by inhibitors of lipid metabolism. Our findings reveal the existence of a novel mechanism of membrane repair that may consist in constriction of the inserted proteinaceous pore within the lipid bilayer. PMID- 10792733 TI - A novel autoregulation mechanism of fnrN expression in Rhizobium leguminosarum bv viciae. AB - The fnrN gene from Rhizobium leguminosarum UPM791 controls microaerobic expression of both nitrogen fixation and hydrogenase activities in symbiotic cells. Two copies of fnrN are present in this strain, one chromosomal (fnrN1) and the other located in the symbiotic plasmid (fnrN2). Their expression was studied by cloning the regulatory regions in lacZ promoter-probe vectors. The fnrN genes were found to be autoregulated: they are expressed only at basal levels under aerobic conditions; they are highly expressed under microaerobic conditions; and they are expressed at basal levels in the double mutant DG2 (fnrN1 fnrN2) under any condition. The promoters of both genes contain two FnrN-binding sequences (anaeroboxes), centred at positions -12.5 (proximal anaerobox) and -44.5 (distal anaerobox). Expression analysis and gel retardation experiments with fnrN1 derivative promoter mutants altered in key bases of the anaerobox sequences demonstrated that binding of FnrN1 to the distal anaerobox is necessary for microaerobic activation of transcription, and that binding of FnrN1 to the proximal anaerobox results in transcriptional repression. The apparent affinity of FnrN1 for the proximal anaerobox was fivefold lower than for the distal anaerobox, resulting in repression of transcription of fnrN1 only at high-FnrN1 concentrations. This positive and negative autoregulation mechanism ensures an equilibrated expression of fnrN in response to microaerobic conditions. PMID- 10792734 TI - PrfA mediates specific binding of RNA polymerase of Listeria monocytogenes to PrfA-dependent virulence gene promoters resulting in a transcriptionally active complex. AB - There is accumulating evidence that the coordinate transcription of the virulence genes in Listeria monocytogenes constitutes a very complex regulation mechanism which might require other factors in addition to PrfA. We previously described an unknown proteinaceous component from crude bacterial cell extracts, which, together with PrfA, formed a specific complex (CI) in electrophoretic mobility shift assays (EMSA) with an hly promoter probe. Here we identify the RNA polymerase (RNAP) of L. monocytogenes as an essential component of the CI complex. Addition of purified RNAP plus PrfA to the hly promoter probe allowed reconstitution of a complex migrating at the same height as CI. By using EMSA and DNaseI footprint experiments it could be shown that PrfA leads to an enhanced and specific binding of RNAP. Transcriptional activity of RNAP in vitro, using the actA promoter, was strictly dependent on PrfA. PMID- 10792735 TI - The use of listeriolysin to identify in vivo induced genes in the gram-positive intracellular pathogen Listeria monocytogenes. AB - Listeria monocytogenes is capable of growth within the cytoplasm of infected host cells. Escape from the host cell phagosome is mediated primarily through secretion of listeriolysin, a haemolytic factor which functions to actively lyse the phagosomal membrane. Listeriolysin negative mutants of L. monocytogenes are non-haemolytic on blood agar plates and demonstrate a significant reduction of virulence in the mouse model of infection. We have developed a system for the identification of in vivo induced genes in L. monocytogenes which utilizes the listeriolysin gene, hly, as both a reporter of gene expression and as a means of selection of promoter elements expressed in vivo. The system is analogous to in vivo expression technology (IVET) first reported for Salmonella, however, as listeriolysin functions in the environment of the host phagosome the loci identified in this study are most likely expressed during residence in the phagosome. The system was successfully tested using the promoter of the inducible virulence gene plcA. A bank was created by fusing a promoterless copy of hly to random promoter elements in a listeriolysin negative IVET host. Sequential inoculations of mice with this bank resulted in the isolation of clones with increased survival potential in the mouse model relative to a negative control, but which remained haemolysin negative on blood agar plates. Nine in vivo induced loci were identified including genes encoding a DNA topoisomerase III, a cellobiose transporter and a fumarase. Two isolates represented fusions to proteins of unknown function and three isolates contained no significant homologues in the database. A mutant in the fumarase gene demonstrated reduced virulence for mice and an inability to grow in cultured mouse phagocytes. PMID- 10792737 TI - Membrane-fusion protein homologues in gram-positive bacteria. PMID- 10792736 TI - Identification of alanine dehydrogenase and its role in mixed secretion of ammonium and alanine by pea bacteroids. AB - N2-fixation by Rhizobium-legume symbionts is of major ecological and agricultural importance, responsible for producing a substantial fraction of the biosphere's nitrogen. On the basis of 15N-labelling studies, it had been generally accepted that ammonium is the sole secretion product of N2-fixation by the bacteroid and that the plant is responsible for assimilating it into amino acids. However, this paradigm has been challenged in a recent 15N-labelling study showing that soybean bacteroids only secrete alanine. Hitherto, nitrogen secretion has only been assessed from in vitro 15N-labelling studies of isolated bacteroids. We show that both ammonium and alanine are secreted by pea bacteroids. The in vitro partitioning between them will depend on whether the system is open or closed, as well as the ammonium concentration and bacteroid density. To overcome these limitations we identified and mutated the gene for alanine dehydrogenase (aldA) and demonstrate that AldA is the primary route for alanine synthesis in isolated bacteroids. Bacteroids of the aldA mutant fix nitrogen but only secrete ammonium at a significant rate, resulting in lower total nitrogen secretion. Peas inoculated with the aldA mutant are green and healthy, demonstrating that ammonium secretion by bacteroids can provide sufficient nitrogen for plant growth. However, plants inoculated with the mutant are reduced in biomass compared with those inoculated with the wild type. The labelling and plant growth studies suggest that alanine synthesis and secretion contributes to the efficiency of N2-fixation and therefore biomass accumulation. PMID- 10792738 TI - Fast tracking the paediatric cardiac surgical patient. PMID- 10792739 TI - Review article: anaesthesia for thoracic surgery in children. PMID- 10792740 TI - Complications of emergency tracheal intubation in severely head-injured children. AB - A high incidence of unsuccessful attempts and complications has been reported when emergency tracheal intubation (ETI) is performed outside the hospital in severely injured children. The aim of this prospective series was to analyse the incidence and related risk factors of complications of emergency tracheal intubation. The time to complete successful ETI and occurrence of incidents, e.g. cough reflex, hypoxia or spasm were related to the experience of the physician performing intubation and the use of drugs to facilitate ETI. The incidence of hypoxia, hypercarbia, postintubation complications such as extubation stridor and long-term sequelae were noted. Of the 188 children, 78% were successfully intubated at the site of the accident, 10% upon arrival at a local hospital from where they were secondarily transferred and 12% upon admission to our trauma centre. The most severely injured children were intubated in the field in 98% of cases without failure, nor life-threatening complications related to ETI. The experience of the operator influenced the number of attempts and the time to complete successful intubation. Immediate incidents were noted in 25% of children, e.g. cough in 18%. The regimen of drugs, but not level of consciousness, influenced the incidence of immediate incidents; without drugs, more than 67% experienced incidents. Early tracheal intubation and controlled ventilation resulted in adequate ventilation upon arrival (mean PaO2 of 35.8+/-24 kPa, mean PaCO2 of 4.35+/-1 kPa). Long-term complications, including transient stridor upon extubation in 33% of the cases, and laryngeal granuloma or tracheal stenosis, were comparable to those in other series. ETI in shocked patients and pulmonary infection in hospital, but not the technique of ETI, increased the risks of long-term complications. Emergency tracheal intubation can be performed safely in the field, and results in adequate ventilation during transportation of severely injured children, provided that it can be performed by trained physicians using adequate drugs to facilitate intubation. PMID- 10792741 TI - Fenoldopam for controlled hypotension during spinal fusion in children and adolescents. AB - Fenoldopam is a dopamine1 agonist whose pharmacological effects include vasodilation of the vascular beds of the kidney, mesentery, skeletal muscle, and coronary systems, resulting in a decrease in systemic vascular resistance and mean arterial pressure. The current retrospective review outlines the use of fenoldopam for controlled hypotension during anterior or posterior spinal fusion in 10 children and adolescents, aged 8-14 years and weighing 22-61 kg. Fenoldopam was infused at a starting dose of 0.3-0.5 microg.kg-1. min-1 and increased incrementally to achieve a mean arterial pressure (MAP) of 50-65 mmHg. The desired MAP was achieved in 4-11 min (7+/-2.5 min). The fenoldopam infusion was administered for 135-225 min (160+/-25 min) in doses ranging from 0.2 to 2.5 microg. kg-1.min-1. The mean fenoldopam infusion rate for the 10 cases varied from 0.5 to 1.4 microg.kg-1.min-1 (1.0+/-0.3 microg.kg-1. min-1). No excessive hypotension or clinically significant adverse effects were noted. Statistically significant, but clinically insignificant, increases in heart rate and decreases in PaO2 were noted during the fenoldopam infusion. The baseline heart rate increased from 87+/-13 b.min-1 to a maximum of 114+/-16 b.min-1 (P < 0.0001) during the fenoldopam infusion. In the six patients undergoing posterior spinal fusion, the baseline PaO2 decreased from 232+/-7 mmHg to a low of 199+/-11 mmHg (P=0.0004) during the fenoldopam infusion. Fenoldopam can be used to provide controlled hypotension during spinal surgery in children and adolescents. Future studies, with direct comparison to other commonly used agents, are needed to better define its advantages and disadvantages as well as its effects on estimated blood loss. PMID- 10792742 TI - Haemodynamic depression by halothane is age-related in paediatric patients. AB - The hypothesis that young infants are more sensitive to the haemodynamic depressant effects of halothane compared with older children was tested. One hundred and sixty unpremedicated, ASA physical status I or II paediatric patients without cardiac or pulmonary disease were divided into five age groups: term neonates, 1-6 months, 6-24 months, 2-6 years and 6-12 years. Anaesthetic induction was achieved with halothane in oxygen and air via mask. Vecuronium 0.1 mg.kg-1 was administered intravenously. During normocapnic manual ventilation by mask, endtidal halothane concentration was maintained at either 2xage-specific MAC (Method I) or 1.7% (Method II) in 20 patients in each age group for 10 min. In both Method I and Method II, systolic and mean blood pressure of term neonates and infants aged 1-6 months decreased significantly (P < 0.01) compared with other age groups. The results of this study demonstrate that neonates and young infants are more susceptible to haemodynamic depression during halothane anaesthesia than are older children, confirming clinical experience. PMID- 10792743 TI - Anaesthesia for manipulation of forearm fractures in children: a survey of current practice. AB - Despite recent recommendations that all children presenting for urgent or emergency surgery should be treated as though they have a full stomach, a local audit had shown a wide variation in technique used for anaesthesia in children after trauma. Therefore, a postal questionnaire was sent to 500 anaesthetists regarding their preferred anaesthetic technique for a 6-year-old child requiring manipulation of a forearm fracture. Four clinical situations were presented differing in the timing of surgery in relation to the injury, starvation times before injury and the administration of opioid analgesia. Rapid sequence induction and tracheal intubation was preferred by 83% of all anaesthetists for surgery on the day of injury if the child had eaten 2 h prior to injury and had received opioid analgesia, but the percentage was significantly lower in experienced anaesthetists (P < 0.05) compared with trainees, and was 34.5% overall if surgery was delayed until the following day. Only 19.3% would perform a rapid sequence induction for surgery on the day of injury if the child had not eaten for 6 h before the injury. We conclude that not all anaesthetists believe that rapid sequence induction is necessary for anaesthesia after forearm fractures, despite recent recommendations. PMID- 10792744 TI - Paediatric lumbar epidurals: a comparison of 21-G and 23-G catheters in patients weighing less than 10 kg. AB - Using the database of our pain management team, we examined the records of 254 patients weighing between 4.5 kg and 10 kg who received an epidural for postoperative analgesia. We looked at the incidence of catheter related problems in two groups of patients in whom either a 21-G (18-G short Tuohy needle) or a 23 G catheter (19-G short Tuohy needle) was used. There was a significantly higher incidence of difficulty in threading the catheter, kinking and occlusion in the 23-G catheter group. There were more leaks in the 21-G catheter group but the difference was not significant and both groups had a comparable incidence of clinically relevant leakages. No dural tap or difficulty in using either needle were reported. PMID- 10792746 TI - Laparoscopic fundoplication in children: anaesthetic experience of 51 cases. AB - We report the anaesthetic management and outcomes of our first 51 laparoscopic fundoplications. Case records of the 50 patients (one redo), median age 6 years (5 months to 20 years), were reviewed. Median duration of anaesthesia was 120 (60 300) min. During the procedure, the heart rate and blood pressure increased by more than 20% over baseline in 18% and 12% cases, respectively. Median increase in PECO2 was 1.0 (0.3-2.3) kPa [7.6 (2.3-18) mmHg]. After surgery, all but one of the patients were managed on a normal surgical ward. Postoperative analgesia requirement was oral or rectal analgesics in 89% of patients and ceased within 48 h of surgery in 95% patients. Median time to discharge home from day of operation was 2 (1-9) days. We conclude that laparoscopic fundoplication in children is well tolerated, there is no requirement for routine postoperative high dependency care and analgesic requirements are minimal. PMID- 10792745 TI - Age-dependent variations in white adipose tissue glycerol and lactate production after surgery measured by microdialysis in neonates and children. AB - In previous studies, we observed that lactate concentrations in interstitial white adipose tissue are higher in small infants than in adults. Moreover, no lipolysis following catecholamine challenge has been reported in neonates and small infants. Our aim was to determine with microdialysis whether the above mentioned age-dependent changes could be detected in situ after surgery. A microdialysis catheter was introduced into the abdominal subcutaneous tissue in 13 neonates and 12 children undergoing surgery. Interstitial concentrations of glucose, lactate and glycerol were measured hourly during the first 20 postoperative hours. The concentrations of lactate in interstitial white adipose tissue were consistently higher in neonates compared to older children, with a significant difference during the 9-18 h postoperative period (P < 0.05). A significant difference in the lactate:glucose ratio was observed at 1-2, 8-10, 15 and 18 h postoperatively (P < 0.05). No significant differences were observed between the two groups with respect to glycerol and glucose concentrations. Interstitial lactate concentrations in white adipose tissue were higher in neonates compared with children in the early postoperative period. No age dependent difference in postoperative lipolysis, measured as interstitial glycerol concentrations, was observed. Thus, an age-dependent difference in interstitial lactate production, but not lipolysis, was detected in the early postoperative period. PMID- 10792747 TI - Increased levels of S-100 protein after cardiac surgery with cardiopulmonary bypass and general surgery in children. AB - The aim of this study was to evaluate changes in concentrations of the neurospecific protein S-100 in relation to cardiac surgery with cardiopulmonary bypass (CPB) and noncardiac general surgery in children below 3 years of age. Seventeen children underwent surgery for congenital heart disease and all survived without clinical signs of neurological complications. Samples for plasma concentrations of S-100 in these patients were taken on three occasions in connection with surgery: before the start of surgery, after CPB and finally 16-20 h after CPB. In the noncardiac group of 31 children, S-100 concentrations were measured on two occasions: before surgery and during surgery. In both groups, a significant increase in S-100 concentrations was observed during surgery, although the increase in the CPB group was significantly higher than in the noncardiac group. The CPB group included four children with Down's syndrome who had higher mean S-100 concentrations on all sampling occasions compared to the remaining patients. The peak S-100 concentrations after cardiac surgery were related to the duration of CPB, the time from the termination of CPB to the first post-CPB sample, as well as mean arterial pressure and cerebral arteriovenous lactate difference during rewarming. All the children studied (Down's patients excluded) had age-dependent plasma concentrations of S-100 measured before surgery. It can be concluded that CPB initiates a marked but transient release of S-100 into the systemic circulation during open heart surgery in children who are not developing clinical signs of neurological sequelae. PMID- 10792748 TI - Analysis of behavioural and physiological parameters for the assessment of postoperative analgesic demand in newborns, infants and young children: a comprehensive report on seven consecutive studies. AB - Many different systems for the assessment of pain in newborns and infants have been tested for validity, rarely for reliability but never for sensitivity or specificity. We aimed to determine whether the assessment of an analgesic demand in the lower age group during the postoperative period is possible by observational methods only. In an repetitive and sequential prospective process for identifying observationable behaviour and measurable physiological parameters as indicators of a postoperative analgesic demand, 584 newborns, infants and young children were studied (7 prospective studies, 4238 observations). Twenty six items were selected as suggested by current literature and for reasons of economy and practicability. The factor analyses resulted in a two-factorial solution with the behavioural items loading on one factor and the physiological parameters on the other (principal component analyses). The physiological parameters blood pressure, respiratory rate and heart rate were found to be unreliable and had no discriminant power to detect an analgesic demand during the postoperative period (discriminant analyses, ROC-curves). In newborns and infants, nine observational items were identified as equally selective, reliable, sensitive and specific to the assessment of postoperative analgesic demand, whereas in young children only five items could be identified (discriminant analyses, ROC-curves). For economic reasons, these five items (crying, facial expression, posture of the trunk, posture of the legs, motor restlessness) were chosen as the basis of an additional pain scale ranging from 0=no pain to 10=maximal (Children's and Infants' Postoperative Pain Scale, CHIPPS). Its internal consistency yielded values for Cronbachs' alpha with 0.92 for toddlers and 0.96 for infants. The coefficient for interrater reliability was 0.93. The scale was validated constructively by the intravenous administration of metamizol, tramadol, nalbuphine, piritramide and ketamine (repeated measures analysis of variance). The Toddler-Preschooler Postoperative Pain Scale and CHIPPS equally identified painfree situations or analgesic demand in 87.4%. In cases with definite pain, the score of CHIPPS was never below 4 points. Seventy one toddlers gave verbal comments on their pain intensity: in 29 painfree situations the CHIPPS score was 3.0 and in 29 painful situations it was 5.7. The values for sensitivity and specificity of CHIPPS were calculated to be 0.92-0.96 and 0.74-0.95, respectively (discriminant analyses). We conclude that it is possible to determine postoperative analgesic demand in the low age group of children by using an observational system such as CHIPPS alone. PMID- 10792750 TI - Anaesthetic management of a 6-week-old child with unilateral pulmonary interstitial emphysema. AB - We report the case of a 6-week-old female infant suffering from respiratory distress related to cystic changes affecting the entire left lung. Anaesthesia was induced with sevoflurane in oxygen and spontaneous ventilation was maintained until intubation of the right main bronchus was secured. A left pneumonectomy was performed and the postoperative course was uneventful. The pathological diagnosis was pulmonary interstitial emphysema. This has not been previously reported in an otherwise normal child delivered uneventfully at term. The anaesthetic management of a child with a cystic lung lesion is discussed. PMID- 10792749 TI - Sedation failures in children undergoing MRI and CT: is temperament a factor? AB - This study examined the relationship between temperament and sedation failure during magnetic resonance imaging (MRI) and computerized tomography (CT). One hundred and two children (aged 3-7 years) who underwent MRI or CT with or without sedation were studied. Demographics, sedatives administered, efficacy of sedation, and adverse events were recorded. Parents completed the Behavioural Style Questionnaire for 3-7 years olds during their wait. Eight children underwent MRI successfully without sedation, 83 were successfully sedated, and 11 procedures were aborted. Children whose sedation failed were less adaptable than children whose sedation was successful (P =0.04). Children who underwent MRI without sedation were more persistent than children who were sedated (P =0.05), and more persistent and less active than those whose sedation failed (P =0.02 and 0.03, respectively). The child's underlying temperament may contribute to sedation outcomes during MRI and CT. A presedation assessment of the child's temperament may therefore assist in decisions regarding sedation. PMID- 10792751 TI - Intramucosal gastric pH during liver transplantation. AB - Intramucosal gastric pH (pHi) was measured during liver transplantation in a 3 year-old girl. The basal value was below that reported as normal for paediatric patients under stable normal ventilatory and haemodynamic conditions. Additional reductions of the pHi were observed during the hepatectomy and anhepatic phase, in the absence of haemodynamic or ventilatory alterations. The reperfusion phase was characterized by a progressive return toward normal values. Because we did not measure oxygen delivery and consumption, we cannot determine whether the reduction of the pHi was due to insufficient oxygen delivery or a result of the vascular anatomical alterations due to the surgical procedure. PMID- 10792752 TI - Dislodgement of bronchial foreign body during retrieval in children. AB - Foreign body aspiration is a leading cause of death in children aged less than 1 year. The removal of a foreign body poses a great challenge to the skill of the anaesthetist. Four cases are presented, analysing the part played by modes of respiration in the dislodgement of a bronchial foreign body during its retrieval. PMID- 10792753 TI - Prolonged somatosensory evoked potential depression following a brief exposure to low concentrations of inhalation anaesthetic in a 3-year-old child. AB - A 3-year-old child was brought to the operating room for removal of a brainstem juvenile pilocytic astrocytoma. Following inhalation induction and intubation, he was maintained on 0.5% isoflurane. Somatosensory evoked potentials (SSEPs) were recorded but unobtainable initially and up to 90 min after all inhalation agents were discontinued. The operation was cancelled and the patient was transported to the paediatric intensive care unit (PICU). Subsequent PICU testing revealed a depression of amplitude with propofol and absence of potentials with 0.5% isoflurane. He returned to the operating room, was induced with propofol, and maintained with a propofol: nitrous oxide:fentanyl technique. This anaesthetic technique allowed adequate tumour resection with appropriate monitoring of SSEPs. These findings suggest that a total intravenous anaesthetic technique may be preferable for resection of spinal cord tumours where SSEPs are monitored. PMID- 10792754 TI - Spinal anaesthesia for insertion of a peritoneal dialysis catheter in a neonate. AB - A 5-day-old neonate born at term presented in renal failure for insertion of a peritoneal dialysis catheter. This was successfully achieved with spinal anaesthesia, with no intra- or postoperative complications. Regional techniques are considered suitable for an increasing variety of procedures in patients in whom the risks of general anaesthesia are significant. PMID- 10792755 TI - Paracetamol-induced fulminant hepatic failure in a child after 5 days of therapeutic doses. PMID- 10792756 TI - The Bair that did not hug. PMID- 10792757 TI - Modification of the LMA no. 1 for diode laser photocoagulation in ex-premature infants. PMID- 10792758 TI - Relative effectiveness of lignocaine-prilocaine emulsion and nitrous oxide inhalation for routine preoperative laboratory testing. PMID- 10792759 TI - Ondansetron - falsely accused? PMID- 10792760 TI - Recombinant chimeric proteins generated from conserved regions of Plasmodium falciparum merozoite surface protein 2 generate antiparasite humoral responses in mice. AB - The merozoite surface protein 2 of P. falciparum is highly polymorphic in nature, but has regions of almost complete conservation at its N- and C-termini. We produced a chimeric recombinant protein comprising these regions only (hereafter termed NC). Mice immunized with the NC antigen produce antibodies at levels comparable to those immunized with 1624, a full-length recombinant protein representing MSP2 from P. falciparum. Antisera raised against NC recognized P. falciparum schizonts by IFA and a P. falciparum protein of Mr 45 kDa by Western blot. However, antibody specificities were observed to differ between anti-NC and anti-1624 sera, and this resulted in differences in parasite recognition, despite similar levels of antibodies having been produced. The response to the NC antigen was also shown to be restricted in some mice (H2-d), but this was overcome by including appropriate T-cell help, which was accomplished by creating recombinant protein chimeras that contained NC and T-helper epitopes from Tetanus toxoid, or MSP119 from P. berghei. PMID- 10792761 TI - Reciprocal cross-protection induced by sporozoite antigens SPAG-1 from Theileria annulata and p67 from Theileria parva. AB - Theileria annulata and Theileria parva both possess a major surface antigen on the sporozoite stage of the life-cycle, called SPAG-1 and p67, respectively. In each case, these antigens are vaccine candidates and have been shown to induce a degree of homologous protection in earlier work. These antigens share sequence homology and are serologically cross-reactive. Here, we confirm that these antigens confer protection against homologous species challenge. More importantly, they mutually confer a degree of cross-species protection raising the prospect of a common vaccine in the future. PMID- 10792762 TI - A new standard for the assessment of disease progression in murine cutaneous leishmaniasis. AB - Infection of mice with Leishmania major has been used both as a model for the cutaneous disease in humans and as a model for the more general control and function of helper T cells in immunity. In both cases, disease patterns and disease progression have been assessed by two complementary methods, lesion size and parasite burden in the draining lymph nodes. We propose a much improved method for the graphical representation of lesion development which conveys more information with better accuracy. We also describe a polymerase chain reaction method for determining parasite burden, which is faster and allows the analysis of larger numbers of experimental animals than the current limiting dilution analysis. Moreover, these methods are equally applicable to other infectious diseases, an obvious one being schistosomiasis. PMID- 10792763 TI - Evaluation of immunization with gut membrane glycoproteins of Ostertagia ostertagi against homologous challenge in calves and against Haemonchus contortus in sheep. AB - Peanut and ConA lectins were used as ligands to isolate glycoproteins from detergent extracts of adult Ostertagia ostertagi membranes. As judged by their profiles following SDS-PAGE, these fractions closely resembled the equivalents from Haemonchus contortus which are derived from the nematode intestinal cell microvillar membranes and which are highly protective when used as antigens. Groups of calves were immunized with the peanut and ConA binding fractions of Ostertagia, either as separate or pooled antigens mixed with QuilA as adjuvant. All calves, including controls immunized with adjuvant only, were challenged with a single dose of infective Ostertagia larvae and faecal egg counts were monitored for 5 weeks. In two experiments where the antigen fractions were pooled, moderate (30-50%), but statistically significant reductions in egg output were observed, but the number of worms was not diminished. No significant protection was observed in a third trial where groups of calves were immunized with peanut or ConA binding proteins given separately. Two further trials were conducted in sheep immunized with the same Ostertagia fractions but challenged with Haemonchus. Irrespective of whether they were administered separately or together, the Ostertagia antigens cross protected efficiently against Haemonchus reducing egg counts by between 81% and 97% and worm numbers by between 57% and 84%. PMID- 10792764 TI - DNA immunization with Onchocerca volvulus genes, Ov-tmy-1 and OvB20: serological and parasitological outcomes following intramuscular or GeneGun delivery in a mouse model of onchocerciasis. AB - The objective of this study was to evaluate whether the distinct immune responses invoked by epidermal and intramuscular DNA immunization could be harnessed to improve upon the levels of protection to Onchocerca volvulus infective larvae achieved previously by recombinant protein immunization. Intramuscular (IM) and epidermal (GeneGun) routes of DNA immunization generally drive T helper1 and Th2 dominant responses, respectively. This dichotomy was used in an attempt to further define the nature of host-protective immunity in a mouse model of onchocerciasis. Mice were immunized with DNA plasmids expressing the O. volvulus antigens, Ov-TMY-1 (tropomyosin) and OvB20 (a nematode specific gene product). While, IM and GeneGun immunization of mice with Ov-tmy-1 induced expected Th1/Th2 associated IgG isotype profiles, mice responded to OvB20 immunization with a Th2 dominant response, irrespective of the delivery route. Despite inducing potent serological responses, neither DNA construct promoted statistically significant levels of protection to L3 challenge infection. We conclude that DNA immunization has good potential for induction of humoral responses against nematode infections and that serological responses alone do not predict vaccination efficacy under the conditions used here to measure host resistance to parasite challenge. PMID- 10792765 TI - Evidence for genomic imprinting of the major QTL controlling susceptibility to trypanosomiasis in mice. AB - Inbred strains of laboratory mice exhibit marked differences in survival time following infection with Trypanosoma congolense, the principal cause of trypanosomiasis in African livestock. The difference in survival time between the relatively resistant C57BL/6 J and more susceptible BALB/c inbred strains has been attributed to three quantitative trait loci (QTLs), Tir1, Tir2 and Tir3. In order to determine whether there was a parent-of-origin effect on this trait, four backcross populations derived from the C57BL/6 J and BALB/c parental strains were bred and inoculated with T. congolense. The two populations with F1 fathers and BALB/c mothers had a significantly greater overall survival rate than the two populations with BALB/c fathers and F1 mothers. This pattern of inheritance suggested the involvement of imprinted genes. Genotyping with markers at the three QTLs controlling susceptibility revealed that the difference in survival time was consistent with genomic imprinting of the QTL of largest effect, Tir1. PMID- 10792766 TI - An animal model for Epstein-Barr virus (EBV)-associated lymphomagenesis in the human: malignant lymphoma induction of rabbits by EBV-related herpesvirus from cynomolgus. AB - It is very important to develop and analyze animal models of Epstein-Barr virus (EBV)-associated tumors in the human. However, only a few reports on the animal models of EBV infection have been reported. Here we review those previous models and describe the details on our newly developed rabbit model of malignant lymphoma induced by EBV-related virus from cynomolgus. In brief, Si-IIA-EBV or Cyno-EBV induced T-cell lymphomas in rabbits inoculated intravenously (77-90%), orally (82-89%), subcutaneously (3/3) and intraperitoneally (2/3) about 2-5 months later. EBV-DNA was detected in peripheral blood by polymerase chain reaction 2 days after oral inoculation of Cyno-EBV while antiviral capsid antigen immunoglobulin G (IgG) was raised 3 weeks after the inoculation. Rabbit lymphomas and their cell lines contained EBV-DNA and expressed EBV-encoded small RNA-1 and EBV-associated nuclear antigen. Rabbit lymphoma cell lines, some of which have specific chromosomal abnormality, showed tumorigenicity in nude mice. The significance and further research subjects of this animal model will be discussed. We believe that the present rabbit model of lymphoma with specific chromosomal abnormalities is very useful for clarifying the role of EBV in human EBV-associated lymphoma and provides a means for studying prophylactic and therapeutic regimens. PMID- 10792767 TI - Fulminant hepatic failure in the elderly: a clinicopathological study of autopsy cases aged over 65 years. AB - The pathology of fulminant hepatic failure (FHF) in the elderly is little known because of its very low frequency. Thirteen autopsy cases, all above 65 years of age (mean +/- SD, 72 +/- 6 years), and 10 younger control cases, all below 40 years of age (30 +/- 7 years), were analyzed. The elderly group comprised 10 cases with subacute FHF and three cases with acute FHF, while the younger group comprised seven cases with subacute FHF and three cases with acute FHF. The most predominant pathological type in the elderly group was submassive hepatic necrosis (10 cases), followed by acute hepatitis with bridging hepatic necrosis (AH-BHN; two cases) and massive hepatic necrosis (one case). In two cases of submassive hepatic necrosis, hepatic regeneration seemed to be insufficient for the suggested history. The underlying diseases and terminal complications were significantly more frequent in the elderly group than in the younger group. In conclusion, the immune response in the elderly group is found to be strong enough to cause massive or submassive hepatic necrosis. However, impaired hepatic regeneration is occasionally observed in the elderly cases and AH-BHN is often lethal because of frequent underlying diseases and severe complications. PMID- 10792768 TI - Autoimmune forms of chronic hepatitis associated with hepatitis C virus (HCV) infection with and without HCV-RNA: histological differences from pure autoimmune hepatitis and chronic hepatitis C. AB - Liver biopsy specimens of pure autoimmune hepatitis (pAIH), autoimmune forms of chronic hepatitis with positivity for anti-hepatitis C virus (anti-HCV) and negativity for HCV-RNA (cAIH-RNA(-)), autoimmune forms of chronic hepatitis with positivity for anti-HCV and HCV-RNA (cAIH-RNA(+)), and chronic hepatitis C (CHC) were compared histologically and statistically to clarify the histological character of the autoimmune form of chronic hepatitis with HCV infection. The following representative histological features were used to investigate: inflammation, fibrosis, plasma cell infiltration, lymphoid aggregates/follicles, non-suppurative destructive cholangitis, and the shape of the enlarged portal tracts. While a considerable overlap in histological features between the pAIH and cAIH-RNA(-) groups and between the CHC and cAIH-RNA(+) groups was recognized, the overlap between the pAIH and CHC groups was small. Significant differences were found between cAIH-RNA(-) and cAIH-RAN(+) groups, especially in necroinflammatory findings. In conclusion, most cases of cAIH-RNA(-) with histological features similar to those of pAIH were shown to be AIH. The remaining cases might be CHC with subsidence of viral duplication. Conversely, many cases of cAIH-RNA(+) with histological findings similar to those of CHC were shown to be CHC clinically mimicking pAIH. The remaining cases might represent coexistence of pAIH and HCV infection. PMID- 10792769 TI - Optically clear nuclei in papillary carcinoma of the thyroid: demonstration of one of the fixation artifacts and its practical usefulness. AB - To examine whether or not optically clear nuclei are one of the fixation artifacts in papillary carcinoma of the thyroid. Experiments using an optimal immersion fixation method by making thin-sliced specimens, 5 x 2 x 2 mm in size were adopted. Fixatives tested were 10% formalin, 10% buffered formalin, mixture of alcohol and formalin (4: 1), 95% alcohol, 4% phosphate-buffered paraformaldehyde, Carnoy's fixative, Bouin's fixative and various concentrations of formalin. Tested thyroid lesions in total were: cases with papillary carcinoma (38); follicular carcinoma (one); follicular adenoma (15); adenomatous goiter (11); Hashimoto's thyroiditis (two); Grave's disease (five); and normal thyroid tissue around tumor (five). Routinely processed papillary carcinoma showed a high occurrence (64.6%) of the optically clear nuclei contrasted by a low occurrence (almost 0%) in other lesions. However, this rate in papillary carcinoma decreased (< 35.3%) in all experimental groups irrespective of the choice of fixatives. Frequent occurrence (61.9-66.6%) of the optically clear nuclei was reproduced in papillary carcinoma fixed in a higher concentration of formalin (40-60%) in the experimental groups. These results demonstrated that the optically clear nuclei are one of the fixation artifacts, although still useful as a diagnostic criterion. PMID- 10792770 TI - A quantitative model using mean and standard deviation for evaluation of interobserver agreement in nuclear atypia scoring of breast carcinomas in a protocol study. AB - A quantitative model was devised for the purpose of evaluating interobserver agreement in the judgment of the nuclear atypia score of cancer cells in protocol studies. In this model, the mean nuclear atypia score (mu) among observers was considered to approximate the 'true' nuclear atypia score of the tumor, and with an identical mu the strength of interobserver agreement was evaluated by the difference between the ideal standard deviation (sigma) and the observed sigma in the slide conference session. This model was applied to the data of a total of 216 invasive ductal carcinomas acquired at eight slide conference sessions held for the standardization of nuclear atypia scoring and monitoring of interobserver agreement level for enrolled cases in a surgical adjuvant protocol on breast cancer. Compared with conventional percentage interobserver agreement and kappa statistics, the present model, named the lambda model, was more sensitive in detecting the improvement of interobserver agreement in nuclear atypia scoring with repetition of slide conference sessions. The lambda model was particularly effective for evaluating the session which included many intermediate cases with a score between 1 and 2 or between 2 and 3. Percentage interobserver agreement and kappa statistics could estimate the agreement irrespective of the number of observers, but the agreement tended to be underestimated when these intermediate cases were in the majority. The present method was considered to be complementary to conventional methods for evaluating interobserver agreement of cancer histology in protocol studies. PMID- 10792771 TI - Atypical hyperplasia of choledocho-pancreatic duct epithelium in an Otsuka Long Evans Tokushima Fatty strain of rats. AB - Epithelial papillary hyperplasia of choledocho-pancreatic duct, associated with cellular atypism, was observed in Otsuka Long Evans Tokushima Fatty (OLETF) rats, a strain originally established as an animal model for non-insulin-dependent diabetes mellitus (NIDDM). To investigate the potential feasibility of OLETF rats as an animal model for pancreatic ductal carcinoma, we examined the pathological characteristics of ductal lesions in OLETF rats aged from 5 to 50 weeks. Hyperplastic lesions in OLETF rats became apparent after 10 weeks of age and increased in severity and frequency of atypical changes in hyperplastic epithelium appearing after 20 weeks. We compared ductal lesions from OLETF rats with those from age-matched Long Evans Tokushima Otsuka (LETO) rats, which share a similar genetic background with OLETF rats but do not develop NIDDM. While LETO rats also display a tendency toward ductal hyperplasia, lesions from OLETF rats were more numerous and larger in size than those from age-matched LETO rats. In addition, lesions from OLETF rats contained a significantly higher number of proliferating cell nuclear antigen-positive cells than those from LETO rats. Finally, lesions in OLETF rats were accompanied by inflammation, and the observed morphological alteration of lesions correlated well with the grade of inflammation. PMID- 10792772 TI - Mosaic-like distribution of endothelial cell antigens in capillaries and juxta alveolar microvessels in the normal human lung. AB - The distribution patterns of endothelial cell antigens, including thrombomodulin and von Willebrand factor (vWf), were studied in normal lung tissues obtained from distant areas of solitary nodules (seven adenocarcinomas and four hamartomas). By single immunoalkaline phosphatase and dual immunofluorescence stainings, the plasma membranes of alveolar capillary endothelium showed linear distribution of thrombomodulin, but their cytoplasm was rarely reactive for vWf (thrombomodulin-dominant pattern). Microvessels with a diameter larger than 10 microm located in the connective tissue zones demonstrated band-like reaction for vWf in their cytoplasm, and their plasma membranes often lacked reactivity for thrombomodulin (vWf-dominant pattern). The juxta-alveolar microvessels located along the borders between the alveolar- and connective-tissue zones showed mosaic like pattern of distribution for these antigens. The pulmonary venules and peribronchial microvessels measuring up to 40 microm in diameter, demonstrated the expression of thrombomodulin along the plasma membrane, and that of vWf in the cytoplasm. Capillaries of the bronchial circulation were also characterized by mosaic-like pattern of distribution. Both antigens were often expressed in a single cytoplasmic segment. The heterogeneous distribution pattern of these antigens suggests topographic difference in endothelial cell function to maintain coagulatory and anticoagulatory balance in the normal human lung. PMID- 10792773 TI - Foregut duplication cyst of the stomach. AB - Foregut duplication cyst of the stomach is an extremely rare disease entity. A 35 year-old Korean man presented with epigastric pain. An abdominal cystic mass, measuring 7 x 6 x 5 cm, was found in the lesser curvature of the stomach. The cyst was unilocular with a grey-white, rubbery wall. Microscopically, the cyst wall was lined by pseudostratified ciliated, columnar epithelium and gastric mucosa with a complete lining of smooth muscle bundles. Although the origin of this lesion remains uncertain, this case suggests that the gastric cyst arose from the embryonic foregut and showed differentiation toward respiratory and gastric structures. PMID- 10792774 TI - Duodenal somatostatinoma: a case report and review of 31 cases with special reference to the relationship between tumor size and metastasis. AB - Somatostatinomas are rare functioning neoplasms usually arising in the pancreas and duodenum. We report a case of somatostatinoma in a 42-year-old male with neither neurofibromatosis nor somatostatinoma syndrome. A large tumor in the descending duodenum had given rise to multiple lymph node metastases. An additional 31 duodenal somatostatinoma cases were also reviewed. Most originated in the descending part of the duodenum, with the ampulla and peri-ampullary area as the most common sites (60%). Frequent manifestations were abdominal pain (25%), jaundice (25%), or cholelithiasis (19%), the latter two reflecting obstruction of the bile duct by tumors. Only two cases showed a possible somatostatinoma syndrome (6%). The tumors with metastases, lymph nodes (10) and liver (2), were significantly larger than average than those without (2.91 +/- 1.49 cm vs 1.36 +/- 0.71 cm, P < 0.05). With a cut-off point of 2.0 cm, diagnostic accuracy for metastasis was 77.78% with 87.50% specificity and 63.64% sensitivity. The smallest tumor with metastases was 0.8 cm and the largest without metastases was 3.0 cm. These results indicate that duodenal somatostatinomas are malignant by nature and the risk of metastasis significantly increases with tumors larger than 2.0 cm. PMID- 10792775 TI - Ossifying fasciitis. AB - A case of ossifying fasciitis occurring in the left femoral region of a 57-year old male is reported. The patient complained of pain in his left lower extremity which was the site of previous catheterization. During a left femoropopliteal by pass, a firm and cylindric mass measuring 5 x 3 x 2.5 cm in size was found and extracted from the left groin. Histologically, a reactional lymph node and an irregularly shaped lesion extended into the perinodal fat tissue which is composed of proliferating fibroblasts with occasional mitotic activity. Within this fibroblastic proliferation, immature woven bone composed of osteoid with calcification and chondroid differentiation were seen. Metaplastic bone is an uncommon finding in cases of nodular fasciitis, parosteal fasciitis, cranial fasciitis and florid reactive periostitis. Ossifying fasciitis is known as an uncommon variant of nodular fasciitis. It is an uncommon post-traumatic benign lesion of subcutaneous tissue with an unclear etiology of ossification which is neither related with a bony structure nor contains any muscle tissue. This lesion has been reported only once previously, in the femoral region after a trauma history of catheterization. PMID- 10792776 TI - Malignant peripheral nerve sheath tumor arising in benign ancient schwannoma: a case report with an immunohistochemical study. AB - A rare example of malignant transformation in an ancient schwannoma arising in the right side of the neck of a 51-year-old man without any clinical manifestations suggesting neurofibromatosis is described. The tumor, approximately 4 cm at its largest dimension, was well circumscribed and had a direct connection with the sympathetic nerve. Microscopically, the central portion of the tumor showed features of ancient schwannoma characterized by extensive hyalinization with cystic degeneration, scattered spindle cells with hyperchromatic and tapered nuclei, and some symplastic changes. However, predominantly in the outer portion, a proliferation of spindle-shaped cells with enlarged nuclei was present. The nuclei of these cells showed irregular contours, coarse granular chromatin texture, and conspicuous nucleoli. Mitotic figures and small necrotic foci with scattered apoptotic bodies were also seen. Immunohistochemically, S-100 protein was almost negative in areas consisting of overtly atypical cells where the mitotic index evaluated with MIB-1 antibody was 30.5%. In contrast, S-100-positive bland spindle cells were scattered in an extensively hyalinized area with a labeling index less than 3%. P53 protein was strongly positive in atypical spindle cells. Although it is a very uncommon event, definite nuclear atypia, frequent mitotic figures, and the existence of small necrotic foci should be recognized as indicating a diagnosis of malignant degeneration of benign schwannoma. Immunohistochemistry would be useful as an ancillary technique in such a setting. PMID- 10792777 TI - Ovarian leiomyosarcoma: an autopsy case report. AB - Primary non-specific sarcoma of the ovary is extremely rare, and only 22 reported cases of pure leiomyosarcoma (LMS) are known to the authors. We present an autopsy case of a primary ovarian leiomyosarcoma in a 73-year-old woman. She had noticed an abdominal mass after difficulty in defecating for several months. The excision of tumor with bilateral salpingo-oophorectomy and hysterectomy was carried out. A diagnosis of pure leiomyosarcoma of the left ovary was made on pathological examination with immunohistochemistry. Adjuvant radio-chemotherapy was not given. At 18 months' follow up, abdomino-pelvic sonography revealed an abdominal tumor and hepatic metastasis. The patient died 3.5 years after the initial surgery. The post-mortem examination revealed a peritoneal recurrent tumor and extensive distant metastases of the liver, lungs, pancreas, gastric mucosa, muscle and skin. The prognosis of the ovarian LMS is poor from the pertinent literature. Several prognostic indicators on histology including mitotic activity, proliferative activity and p53 status of the tumor are discussed. PMID- 10792778 TI - Quantitative estimation of renin-containing cells in the juxtaglomerular apparatus in Bartter's and pseudo-Bartter's syndromes. AB - Our colleagues have previously observed the number of renin-immunoreactive juxtaglomerular apparatus (JGA) in renal specimens from patients with Bartter's and pseudo-Bartter's syndromes and from the normal individual. The question we have is to what extent the renin-containing cells actually increased their cell number. It is easy to count the number of renin-positive JGA in the specimens. The data showed that the number of renin-positive glomeruli was 10-17 times more frequent in both syndromes than in the normal control. A geometrical calculation simply indicated that the probability of detection of a cell mass in a thin cross section is proportional to the length of the target structure. The number of renin-containing cells per JGA is proportional to the increased rate of detection of renin-immunoreactive JGA per unit area, namely 10-17 times. This is true when we assume that renin-containing cells are solely located along the afferent arteriole, where differentiation toward renin-secreting cell occurs in a few layers just outside the arteriole in a longitudinal direction. PMID- 10792779 TI - Review article: acute inorganic mercury vapor inhalation poisoning. AB - Mercury contamination is a serious environmental problem worldwide. Two primary sources of contamination are dumping of large quantities of inorganic mercury and exposure in the mining industry. Although the actual fatal level of mercury vapor is not known, exposure to more than 1-2 mg/m3 of elemental mercury vapor (Hg0) for a few hours causes acute chemical bronchiolitis and pneumonitis. Two hours after exposure, lung injury appears as hyaline membrane formation, and finally, extensive pulmonary fibrosis occurs. Clinical findings correlate with the duration of exposure, the concentration of mercury, and the survival time after exposure. There is no correlation between pathological findings and the concentration of mercury in the tissues. Necrosis of proximal convoluted tubules may be attributed to the disruption of the enzyme systems of Hg2+-sulfhydryl compounds. Metallothionein protein (MT), induced by the accumulation of Hg2+ in the kidneys, may play an important role in detoxication after it forms a non toxic Hg2+-MT compound. Despite the deposition of mercury in the brain, compared with organic mercury, inorganic mercury did not seem to damage the neurons. Drugs such as chelating agents and corticosteroids appear to effectively decrease the inflammation and delay pulmonary fibrosis. PMID- 10792780 TI - Time-specific occurrence of alopecia in neonatal C57BL mice treated with N-methyl N-nitrosourea and the therapeutic efficacy of tacrolimus hydrate. AB - Alopecia was induced in male and female neonatal C57BL mice by a single intraperitoneal injection of 60 mg/kg N-methyl-N-nitrosourea (MNU). MNU administration was most effective in the 8-day-old mice and less effective in the 5-day-old mice (at active and early anagen stages of the first hair cycle, respectively). No alopecia was seen in the day 14 MNU-treated animals (at telogen stage of the first hair cycle). MNU effectively induced hair follicular cell apoptosis at the anagen stage by up-regulation of Bax protein without down modulation of Bcl-2 protein. In day 8 MNU-treated mice, the immunosuppressive agent 0.01% tacrolimus hydrate (FK506), when topically applied for 5 days from 1 day after MNU treatment (before the occurrence of alopecia), decreased the severity of alopecia. However, it did not stimulate hair growth when applied for 5 days from 20 days of age (after occurrence of alopecia). PMID- 10792781 TI - Fine mapping of thymus enlargement gene 1 (Ten1) in BUF/Mna rats. AB - Two polymeric autosomal loci, Ten1 and Ten2, regulate thymus enlargement in BUF/Mna (B) rats. Previously, we mapped Ten1 on chromosome (Chr) 1 to a 20 cM region between Myl2 and D1Mgh11, and Ten2 on Chr 13. To further characterize the precise position of Ten1, 34 and 37 microsatellite markers, that have a polymorphism between the B and WYK (W) and between the B and MITE (M) strains, were used for linkage analysis of thymus enlargement in 105 (WBF1 x B) blackcross (BC) and 78 (B x BMF1) BC rats, respectively. Our data showed that the D1Rat168, D1Rat112, D1Rat323, D1Got186, D1Got187 and D1Got188 markers each gave a peak logarithm of odds (LOD) score of 10.68 for linkage to the thymus ratio in (WBF1 x B) BC rats, and that the D1Rat168, D1Rat197, D1Got184, D1Got186 and D1Got188 markers each gave a peak LOD score of 7.82 in (B x BMF1) BC rats. The two LOD score peaks are coincident in the position of the rat genetic map. All of the markers mentioned above are located in the region between Igf2 and D1Mgh11, in which synteny is conserved with human 11q15.5 and the distal end of mouse Chr 7 or with human 11q13 and the proximal end of mouse Chr 19. Genes existing in these regions are discussed as candidate genes for Ten1. PMID- 10792782 TI - Activation of the TCL1 protein in B cell lymphomas. AB - The TCL1 gene, localized near the break point of chromosome 14q32.1 often involved in T cell leukemias, is also expressed in normal precursor T and B cells, and B cell lymphoma cell lines. We investigated the expression of the TCL1 protein in various types of B cell lymphomas according to the Revised European American Classification of Lymphoid neoplasms. Paraffin-embedded tissue sections of lymphoma specimens were subjected to TCL1 immunohistochemistry, and positivity was scored on a three-tiered scale: - (< 25% cells), + (25-50% cells), and ++ (> 50% cells). The TCL1 protein was expressed in low-grade B cell lymphomas including mucosa-associated lymphoid tissue type in ocular adnexa (18/20, 90%). It was also expressed in follicular, lymphoplasmacytic, and mantle cell lymphoma, but not in high-grade diffuse large B cell lymphoma (2/11, 18%). These data suggest that the expression of the TCL1 gene characterizes low-grade B cell lymphomas, and may be involved in certain processes of lymphomatogenesis. PMID- 10792783 TI - Semiquantitative immunoblot analysis of nm23-H1 and -H2 isoforms in adenocarcinomas of the lung: prognostic significance. AB - Total amounts of nm23 protein and relative levels of H1 and H2 isoforms were studied in 27 fresh-frozen samples of pulmonary adenocarcinoma and adjacent non neoplastic tissues that were obtained at surgery. Semiquantitative immunoblotting with a monoclonal antibody (Pan-242) against nm23 protein demonstrated both isoforms, recognized as 20.5 kDa for H1 and 18.5 kDa for H2, to be present in all cases. Both H1 and H2 levels in neoplastic tissues were higher than in the corresponding non-neoplastic samples. Expression of H2 was usually greater than of H1. The H2/H1 ratio varied from 1.9 to 14.1 (mean value 5.2) in non-neoplastic tissues and 1.0-5.9 (mean value 2.5) in neoplastic tissues, although this ratio did not correlate with any prognostic factor like tumor size, nodal status or distant metastasis (TNM tumor stage). H1 and H2 levels were significantly lower (mean values 4.3 and 2.4) in well-differentiated than in moderately and poorly differentiated adenocarcinomas (8.3 and 3.0) (P < 0.03 and P < 0.05, respectively). These data indicate that H1 and H2 isoform levels correlate with histological differentiation, but not the metastatic potential or stage of pulmonary adenocarcinoma. PMID- 10792784 TI - Quantitative analysis of apoptotic cell death in granulomatous inflammation induced by intravenous challenge with Cryptococcus neoformans and bacillus Calmette-Guerin vaccine. AB - Apoptotic cell death of macrophage has become recognized as a significant mechanism responsible for the resolution of inflammation. The purpose of this study was to examine how the apoptotic cell death involves the formation and resolution of granulomas in rats intravenously inoculated with Cryptococcus neoformans (Cr. neoformans) and Mycobacterium bovis-derived bacillus Calmette Guerin (BCG) vaccine. The number and size of granulomas in the livers obtained on days 5, 10, 15, 20 and 25 after inoculation were examined by morphometric image analysis, as well as the occurrence of apoptotic cell death quantitatively analyzed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL) procedure on tissue sections. In both groups the number and size of granulomas were maximized on day 10, then the granulomas were almost resolved until day 25 when the inoculated Cr. neoformans and BCG almost disappeared. From the induction to the resolving stages of granulomatous inflammation, TUNEL-positive cells constantly appeared in granulomas, and the highest frequency of apoptotic cells in granulomas was observed in the earlier stage of granuloma formation. These results indicate that the maintenance and resolution of infectious granulomas are regulated by the balance between the influx of newly recruited macrophages and the apoptotic elimination of granuloma macrophages. The apoptosis of granuloma macrophages actively involves the cellular turnover in both granuloma formation and resolution. PMID- 10792785 TI - Incidence of P53 and K-ras alterations in ovarian mucinous and serous tumors. AB - Clarification of the pathogenic relationships existing among ovarian cystadenomas, tumors of low malignant potential (LMP) and various adenocarcinoma types, a series of 29 mucinous and 19 serous ovarian tumors including adenomas, LMP tumors and adenocarcinomas were examined. P53 protein was detected by the streptavidin-biotin method and point mutation of K-ras codon 12 was detected by polymerase chain reaction-restriction fragment length polymorphism analysis. P53 overexpression was observed more frequently in serous adenocarcinomas (5/8, 63%) than in mucinous adenocarcinomas (2/9, 22%) and was correlated with the malignant potential of serous tumors. Furthermore, the proportion of P53-positive cells was significantly higher in serous adenocarcinomas than in mucinous adenocarcinomas. P53 overexpression may therefore be closely related to the early events of carcinogenesis in serous tumors. Although mutation of the K-ras oncogene appears to be an important event in the early tumorigenesis of mucinous tumors, mutation of the K-ras oncogene in serous tumors may be dependent on morphology. Different complex pathways of oncogene and/or tumor suppressor gene abnormalities may be involved in the development of mucinous and serous adenocarcinomas. PMID- 10792786 TI - Placental pathology in systemic lupus erythematosus with antiphospholipid antibodies. AB - Systemic lupus erythematosus (SLE) is associated with a poor pregnancy outcome. Antiphospholipid antibodies (APL), which include lupus anticoagulant (LAC) and anticardiolipin antibodies (aCL), are frequently found in patients with SLE, and their presence has been associated with fetal loss. To examine placental pathologic features of SLE patients with APL, we performed a pathologic study on 47 placental tissue samples from 47 pregnant SLE patients with APL (15 patients; four LAC single-positive patients, seven aCL single-positive patients, four LAC and aCL double-positive patients) and without APL (32 LAC and aCL double-negative patients). The incidence of extensive infarction, decidual vasculopathy, decidual thrombosis and perivillous fibrinoid change, which have been thought to be characteristic lesions of APL placenta, was significantly higher in the LAC and aCL double-positive patients than in the patients without APL. Conversely, the above-mentioned lesions between the LAC or aCL single-positive patients and the APL negative patients did not differ significantly. Among the 15 patients with APL, two of the three patients with both decidual vasculopathy and thrombosis had extensive infarction associated with fetal death. Moreover, the patients having fetal death showed LAC and aCL double-positivity. In conclusion, this study indicated that the LAC and aCL double-positivity is an important factor for extensive infarction resulting from decidual vasculopathy and decidual thrombosis in the SLE placenta. Moreover, it was indicated that LAC and aCL double positivity is an important risk factor for fetal death in the SLE patient. PMID- 10792787 TI - Abnormal pigmentation of schwannoma attributed to excess production of neuromelanin-like pigment. AB - Five cases of non-melanotic pigmented schwannoma with excess accumulation of neuromelanin are presented. The tumors were composed basically of spindle or fusiform tumor cells, compatible with those of classical schwannoma, together with varying numbers of tumor cells containing various amounts of light brown or grayish pigment. Fontana-Masson stain demonstrated argentaffin granules in some tumor cells of each tumor and bleaching with potassium permanganate abolished argentaffin reaction. Ultrastructural examination demonstrated the granules contained fine particles with heterogeneous density, occasionally together with coarse granular materials or amorphous high-density areas, indicating lysosome or autophagosome. Neither typical melanosomes nor neurosecretory granules were detected. In immunohistochemistry, neurogenic markers as well as CD68 were expressed in most tumor cells in each case and various numbers of tumor cells were positive for Leu7 and CD34. Lysozyme was also frequently positive in tumor cells, especially in granular cells. HMB45 was not expressed in any of the cases. These findings indicate that these cases are schwannomas with abnormal accumulation of neuromelanin-like pigment. PMID- 10792788 TI - Myoepithelioma of the lacrimal gland: report of a case with potentially malignant transformation. AB - Myoepithelioma of the lacrimal gland is extremely rare and only four cases, one of which was malignant, have been reported in detail. The present report describes a case of lacrimal gland myoepithelioma in a Japanese male with histological features suggestive of potentially malignant transformation. The excised tumor consisted of two components, a central nodular component and a peripheral component surrounding the former. These components were separated by a fibrous tissue. Microscopically, both components were comprised almost entirely of spindle-shaped cells, but with some epithelioid cells containing glycogen granules. Extracellular spaces in the peripheral component were filled with eosinophilic materials with the occasional crystalloid structures, which were immunoreactive for collagen type I. Neoplastic cells were immunoreactive focally for vimentin and S-100, but negative for cytokeratins, epithelial membrane antigen, muscle actin, smooth muscle actin, desmin, myosin, and glial fibrillary acidic protein. The neoplastic cells in the central component showed nuclear pleomorphism and atypia with a higher frequency of mitotic figures, and higher labelings of proliferation markers than those in the peripheral component. Neither invasion, necrosis, nor hemorrhage was observed in the tumor. From these findings we proposed a diagnosis of potentially malignant myoepithelioma. PMID- 10792789 TI - Adrenal rest tumor of the liver: a case report with immunohistochemical investigation of steroidogenesis. AB - A case of adrenal rest tumor arising in the liver of a 62-year-old male with chronic hepatitis type C is reported. The tumor was clinically non-functioning and required distinction from hepatocellular carcinoma. The yellowish-brown tumor measured 25 x 18 x 15 mm and was located in the subcapsular portion of the right hepatic lobe. Histologically, the tumor presented features similar to those of the adrenal cortex and was predominantly composed of pale cells. Electron micrograph revealed lipid droplets and mitochondria with tubulo-vesicular cristae, consistent with the characteristics of steroid-producing cells. Immunohistochemically, the tumor expressed the adrenal 4 binding protein and a number of enzymes involved in the synthesis of adrenocortical steroids. At surgery, the right adrenal gland was present independently from the liver. This hepatic tumor was considered to be an adrenal rest tumor with steroidogenic capability. PMID- 10792790 TI - Adult-onset giant juxtaglomerular cell tumor of the kidney. AB - The juxtaglomerular cell tumor (JGCT) of the kidney is a rare neoplasm which commonly secretes renin. This tumor often occurs in teenagers. This paper documents the 14th adult-onset (over 30-years-old) case with a giant JGCT which measured 9.0 x 8.0 x 7.5 cm. Histologically, this tumor was composed of both vascular and tubular components. Immunohistochemically, the vascular component reacted with renin, cytokeratin 7, ulex europaeus agglutinin-1, vascular endothelial growth factor (VEGF) and Flk-1 (VEGF-R2), whereas the tubular component was positive for renin, epithelial membrane antigen (EMA), cytokeratin 7, alpha-1-antitrypsin, VEGF and Flk-1. This finding suggests that both vascular and tubular components of JGCT may promote neoplastic proliferation via an autocrine mechanism through the action of VEGF. PMID- 10792791 TI - Solitary infantile myofibromatosis in the femur. AB - We report on a case of the solitary type of infantile myofibromatosis of the right femur in an 11-month female. Radiographically, a well-defined osteolytic lesion was seen in the diaphysis of the right femur. Pathological study revealed that the spindle-shaped cells with eosinophilic cytoplasm were arranged in a fascicular and intertwining fashion. The cleft-shaped vascular spaces were observed between tumor nodules. Immunohistochemical staining caused many tumor cells to react for vimentin, alpha-smooth muscle actin and desmin. Ultrastructural study revealed numerous thin and intermediate types of filaments in the cytoplasm of the cells. To our best knowledge, this is the third reported case of solitary infantile myofibromatosis of long bones. PMID- 10792792 TI - Intraplacental choriocarcinoma with fetomaternal transfusion. AB - Intraplacental choriocarcinoma is very rare, and is usually found only after maternal and fetal metastatic disease is identified. The purpose of this case report is to review the incidence and findings of intraplacental choriocarcinoma. A term placenta was investigated because the newborn was born with severe anemia (Hb 3.0 g/dL). A 2 cm nodule was noted on the surface of the amniotic membrane and grossly resembled an infarction. The tumor was examined microscopically with immunohistochemical staining for the alpha- and beta-human chorionic gonadotropin (alpha-hCG, beta-hCG) subunits, human placental lactogen (hPL) and Ki-67. Microscopically, the tumor consisted of necrotic areas with proliferation of atypical trophoblastic cells and destruction of the villi and capillaries. The cells were positive for the alpha-hCG, beta-hCG subunits, hPL and Ki-67, consistent with intraplacental choriocarcinoma. The mother and newborn were investigated for the presence of metastatic disease. Computed tomography scans and magnetic resonance imaging of the mother and infant were negative for metastatic disease. Choriocarcinoma, limited only to the placenta with no evidence of metastatic disease is very rare. Primary intraplacental choriocarcinoma may frequently be overlooked or missed, and choriocarcinoma may possibly arise in the placenta more often than in retained or persistent trophoblast following pregnancy. PMID- 10792793 TI - High-dose systemic corticosteroids can arrest recurrences of severe mucocutaneous erythema multiforme. AB - We treated two children with a total of five recurrences of erythema multiforme major with pulsed dose, systemic corticosteroid therapy on an open basis, and report herein the beneficial effects of this regime. We mean to further reevaluate this therapy on a controlled study basis. PMID- 10792794 TI - Cutaneous markers of primary immunodeficiency diseases in children. AB - Primary immunodeficiency diseases (PIDs) are rare but important conditions found predominantly in children. We studied PIDs in a large pediatric hospital, their association with cutaneous alterations, and the importance of cutaneous alterations as diagnostic markers. Among 382,383 pediatric patients, 130 (0.0003%) had a PID: humoral in 27, cellular and combined in 18, phagocytic in 37, and associated with major defects in 45. An average of two cutaneous alterations were present in 90 (69%) patients: infections in 80, eczema dermatitis in 38, and miscellaneous in 57. In 71 (79%) patients the cutaneous alterations preceded and were the basis for the clinical immunologic diagnosis. Only two PIDs were not associated with cutaneous lesions. PMID- 10792795 TI - Childhood pernio and cryoproteins. AB - Childhood pernio is an uncommon condition described mainly through isolated case reports. We examined the cutaneous spectrum, clinical associations, presence of cryoproteins, and evolution of the condition in children, and performed a retrospective case series evaluation of children with pernio seen at a single ambulatory care university center over a 10-year period. Cases were drawn from a population of 3.2 million. Follow-up was at least 3 years. We found four boys and four girls with pernio. Distribution of skin lesions was on the fingers, toes, and ears. Four children had cryoglobulins or cold agglutinins, two had a positive rheumatoid factor, and none had a positive ANA or ANA profile. All eight cleared within 3 months and did not recur over at least a 3-year period. We concluded that childhood pernio is uncommon and may be associated with the presence of cryoproteins. PMID- 10792796 TI - Cutis marmorata telangiectatica congenita: clinical findings in 85 patients. AB - Cutis marmorata telangiectatica congenita (CMTC) is an uncommon, sporadic, congenital cutaneous condition presenting with persistent cutis marmorata, telangiectasia, phlebectasia and possible ulceration of the involved skin, skin atrophy, and undergrowth of the involved extremity. To further the current understanding of this disorder, we analyzed the clinical features of a large series of patients with CMTC. The files of 85 patients with CMTC who were examined in our center over the last 20 years were reviewed. The data recorded included patient sex, age at onset, nature and distribution of the lesions, associated abnormalities, and course of disease. CMTC was characterized by early recognition of the lesions (94% at birth) and equal sex distribution. The lesion was unilateral in 65% of patients and most commonly involved the limbs (69%). Additional capillary lesions, noted in 20% of patients, may well have been an extension of the CMTC itself. Associated anomalies were found in 18.8% of patients. Almost half of the patients showed a definite improvement in the reticular vascular pattern on follow-up. No familial cases of CMTC were noted. CMTC is more prevalent than previously recognized, affects both sexes equally, and has a mostly localized distribution; additional vascular lesions are associated with the disorder, but other anomalies occur less often. Prognosis is generally good. PMID- 10792797 TI - Scleroderma-like indurations involving fascias: an abortive form of congenital fascial dystrophy (Stiff skin syndrome). AB - Four patients are described with stone-hard indurations of the skin and subcutaneous tissue, predominantly on the buttocks and thighs, in the areas of the thickest fascia lata and glutealis. All cases were sporadic, started in early infancy, were only slightly or not progressive, and showed no visceral involvement or immunologic abnormalities. In all, the hallmark of the disease was strikingly enlarged fascia. In one patient, typical features developed progressively for 9 years, and in two patients the changes remained abortive, limited to some areas, and not symmetrical. The fourth patient showed some similarity to profound morphea with no cutaneous involvement. Recognition of atypical or abortive cases of congenital fascial dystrophy, which is probably a variant of heterogeneous stiff skin syndrome involving exclusively fascia, is of practical importance, since no therapy is required. However, intensive rehabilitation should start in early infancy and continue throughout life. The genetic defect of molecular organization of collagen in the fascia results in formation of giant amianthoid-like collagen fibrils. PMID- 10792798 TI - Staphylococcal septicemia in children with atopic dermatitis. AB - Atopic dermatitis (AD) is frequently complicated by minor bacterial superinfections. Invasive infections such as osteomyelitis have rarely been reported. We describe two children with staphylococcal septicemia during an exacerbation of their AD. Cellulitis and underlying congenital heart disease, respectively, were considered predisposing factors for the development of bacteremia. Identical strains were isolated from the skin, and there was a significant increase in antibodies against Staphylococcus aureus capsular polysaccharide in one child. Our cases demonstrate the potential severity of bacterial skin infections in AD, especially when associated with an underlying condition that increases vulnerability to bacteremia. While their true incidence in children with AD is currently unknown, it is conceivable that systemic staphylococcal infections may be more common than previously thought. Staphylococcal bacteremia has to be considered in the differential diagnosis of fever in children with severe AD. Conversely, episodes of staphylococcal bacteremia should prompt a search for underlying predisposing factors. PMID- 10792799 TI - KID syndrome associated with features of ichthyosis hystrix. AB - Keratitis-ichthyosis-deafness (KID) syndrome is a congenital ectodermal disorder causing erythrokeratoderma, vascularizing keratitis, and neurosensory deafness. Ichthyosis hystrix is a rare cutaneous disease characterized by well-demarcated, spiky, verrucous, linear plaques that is believed to be a clinical and pathologic chimera of two autosomal dominant diseases: epidermal nevus and epidermolytic hyperkeratosis. We present a patient with the classic triad of KID syndrome with clinical and histologic features of ichthyosis hystrix. This case demonstrates that KID syndrome comprises a spectrum of ectodermal disorders which may include diseases such as hystrix ichthyosis and deafness (HID) syndrome. PMID- 10792800 TI - Congenital midline cervical cleft: case report and review of the English language literature. AB - Congenital midline cervical cleft is a rare anomaly of the ventral neck that heretofore has not been reported in the dermatology literature. We present a case of a midline cervical cleft that was diagnosed and managed at an early age. We also review the literature and discuss its clinical and histologic features, treatment, and possible embryology. PMID- 10792801 TI - Disfiguring draining sinus tracts in a female acne patient. AB - A 17-year-old girl with a 9-month history of papulopustular acne developed disfiguring, highly inflammatory, fluctuant nodules in both nasolabial folds within 2 months. Periodically she experienced discharge of pus and blood from these lesions. A diagnosis of disfiguring draining sinus tracts associated with acne vulgaris was made. The therapeutic regimen included intralesional corticosteroid injection, systemic corticosteroids along with a macrolide antibiotic, and systemic isotretinoin to reduce the inflammatory process. Outcome was favorable, with no recurrences during the following 10 months. Draining sinus is a malevolent lesion usually seen in severe forms of acne such as acne conglobata, acne fulminans, and acne inversa. Treatment is difficult and often unsatisfactory. In many cases, excision of the lesion is necessary to provide a permanent cure. PMID- 10792802 TI - Auriculotemporal (Frey) syndrome in late childhood: an unusual variant presenting as gustatory flushing mimicking food allergy. AB - Auriculotemporal or Frey syndrome is characterized mainly by recurrent episodes of facial gustatory flushing and/or sweating, limited to the cutaneous distribution of the auriculotemporal nerve. Although relatively common in adults following injury to the auriculotemporal nerve or parotid disease, the condition has rarely been reported in children. Moreover, in childhood, auriculotemporal syndrome has been described mainly in infancy and early childhood as a sequel of perinatal birth trauma resulting from assisted forceps delivery. We report a 13 year-old girl with a 2-month history of recurrent, painless, preauricular gustatory flushing without sweating, initially suspected to be a food allergy. Detailed inquiry revealed a history of a bicycle accident with mandibular condyle fracture 7 years prior to the onset of symptoms. Our patient demonstrates an unusual presentation of auriculotemporal syndrome in late childhood as gustatory flushing mimicking food allergy. Awareness of this variant is essential for prompt recognition, thus avoiding unnecessary laboratory tests, especially as this condition usually resolves spontaneously. PMID- 10792803 TI - Myxoid liposarcoma in a 12-year-old girl. AB - A 12-year-old girl developed a myxoid liposarcoma on the inner side of her right thigh. Liposarcomas are extremely rare at this age compared to benign lipoblastomas, which are the most frequent tumors of fatty tissue in childhood. The prognosis of myxoid liposarcoma is good, especially when, as in this patient, the tumor is located in the subcutis where a large and deep excision is possible and often curative. PMID- 10792804 TI - Disseminated lupus vulgaris and papulonecrotic tuberculid: case report. AB - The incidence of tuberculosis and extrapulmonary forms of this disease is increasing all over the world. Lupus vulgaris is the most prevalent form of cutaneous tuberculosis in Europe and the Middle East. Papulonecrotic tuberculid, the most common form of hyperergic response to mycobacteria or their fragments, is uncommon in children. We report lupus vulgaris with papulonecrotic tuberculid in a 12-year-old girl who had a 3-year history of slowly growing plaques on her trunk, extremities, and the tip of her nose and papuloulcerative lesions over her entire body. A skin biopsy specimen showed minimally caseating granulomatous inflammation. Staining for acid-fast bacilli was negative in both plaques and papules. Polymerase chain reaction identified Mycobacterium tuberculosis DNA in the patient's sputum, gastric fluid, and plaques and was negative in the papules. She was started on antituberculous therapy with four drugs and her lesions responded rapidly. PMID- 10792805 TI - Congenital linear steatocystoma multiplex of the nose. AB - We describe a case of steatocystoma multiplex with an unusual clinical manifestation. A 5-year-old girl had 8 papulonodular lesions arranged in a linear fashion on the left side of the nose. These had been present at birth. Histologically the lesions showed the typical features of steatocystoma multiplex. There was no family history of similar lesions. This is the second case of linear steatocystoma multiplex, a rare variant of steatocystoma multiplex. PMID- 10792806 TI - Henoch-Schonlein purpura: simultaneous occurrence in two siblings. AB - Simultaneous occurrence of Henoch-Schonlein purpura (HSP) in family members is not well documented. We describe simultaneous onset of HSP in two sisters 1 day after the wearing of new synthetic slippers. Such an occurrence of the disease implies a common cause, however, in most patients the search for a causative agent is usually futile. There was no clear evidence of infection in our patients. The association of the appearance of the disease with the use of the slippers in our patients could indicate a possible, although unlikely, cause for their HSP. PMID- 10792807 TI - Granuloma gluteale infantum: a case report. AB - Granuloma gluteale infantum is a benign disorder with the clinical appearance of "granulomatous" nodules involving the diaper area. It is an uncommon disorder considered a complication of primary irritant diaper dermatitis. A 6-month-old boy with reddish-purple oval nodules located on the right inguinal fold and right buttock diagnosed as granuloma gluteale infantum is presented. A contributing factor in our patient could be a primary irritant dermatitis or a preexisting candidial infection. The name granuloma gluteale infantum is a misnomer since no granulomas are found in these lesions. PMID- 10792809 TI - A scaly macule on the bridge of the nose of a 15-year-old boy. PMID- 10792808 TI - Crusted scabies in an immunocompetent child: treatment with ivermectin. AB - An 11-year-old girl presented to our clinic with recalcitrant crusted scabies despite repeated applications of topical scabicides. She had no history of corticosteroid use prior to onset of the eruption and no evidence of immunodeficiency. A combination of oral ivermectin, topical lindane, and keratolytics cleared the infestation. Our patient is exceptional in that she had no risk factors commonly associated with a propensity to develop crusted scabies. While topical therapy remains the first-line treatment for children with classic scabies, in the unusual instance of a child with recalcitrant, crusted scabies, ivermectin may offer an efficacious alternative, although it should be used with caution. We discuss the use of oral ivermectin for treatment of crusted scabies and the challenging comprehensive management needed for this socially stigmatizing condition. PMID- 10792810 TI - What syndrome is this? Killian-Teschler-Nicola syndrome. PMID- 10792812 TI - Phakomatosis pigmentovascularis type IIA in a Caucasian child. PMID- 10792811 TI - Scabies therapy for the millennium. PMID- 10792813 TI - Treatment of linear IgA bullous dermatosis of childhood with colchicine: in reply. PMID- 10792814 TI - Epidermolysis bullosa acquisita in a 4-year-old boy. PMID- 10792815 TI - Transactivation properties of parsley proline-rich bZIP transcription factors. AB - Light-responsive chalcone synthase (CHS) gene activation requires LRUCHS, a light regulatory promoter unit including the MYB recognition element MRECHS and the ACGT-containing element ACECHS. ACECHS is bound by the parsley basic region/leucine zipper (bZIP) factors CPRF1 and 4. Factors containing the bZIP domain exist in animals, plants and yeast, and recognize DNA sequence specifically after formation of homo- or heterodimers. To determine the potential role of CPRFs in the regulation of CHS promoter activity, we investigated the functions of distinct CPRF domains in a homologous co-transfection system. The proline-rich domains of CPRF1 and CPRF4 activate transcription, indicating that CPRF1 and CPRF4 have transactivating properties. Over-expression of the CPRF1 bZIP domain caused a reduction of LRUCHS-mediated light inducibility, and point mutations throughout ACECHS affected both responsiveness to UV-containing white light and transactivation by CPRF1:VP16. The data suggest that a CPRF1-containing bZIP heterodimer interacts with ACECHS in vivo. We discuss regulatory steps in light-induced CHS transcription that may be influenced by CPRF1 and/or related bZIP factors. PMID- 10792816 TI - Ion currents involved in early Nod factor response in Medicago sativa root hairs: a discontinuous single-electrode voltage-clamp study. AB - Nod factor [NodRm-IV(Ac,S)], isolated from the bacterium Rhizobium meliloti, induces a well-known depolarization in Medicago sativa (cv Sitel) root hairs. Analysis of this membrane response using the discontinuous single-electrode voltage-clamp technique (dSEVC) shows that anion channel, K+ channel and H+ ATPase pump currents are involved in young growing root hairs. The early Nod factor-induced depolarization is due to increase of the inward ion current and inhibition of the H+ pump. It involved an instantaneous inward anion current (IIAC) and/or a time-dependent inward K+ current (IRKC). These two ion currents are then down-regulated while the H+ pump is stimulated, allowing long-term rectification of the membrane potential (Em). Our results support the idea that the regulation of inward current plays a primary role in the Nod-factor-induced electrical response, the nature of the ions carried by these currents depending on the activated anion and/or K+ channels at the plasma membrane. PMID- 10792817 TI - Functional analysis of HD2 histone deacetylase homologues in Arabidopsis thaliana. AB - Post-translational modification of histones, in particular acetylation, is an important mechanism in the regulation of eukaryotic gene expression. Histone deacetylases are enzymes that remove acetyl groups from the core histones and play a key role in the repression of transcription. HD2 is a maize histone deacetylase, which shows no sequence homology to the histone deacetylases identified from other eukaryotes. We have identified two putative HD2-like histone deacetylase cDNA clones, AtHD2A and AtHD2B, from Arabidopsis thaliana by screening the expressed sequence tag database. AtHD2A and AtHD2B encode putative proteins of 246 and 305 amino acids, and share 44% and 46% amino acid identity to the maize HD2, respectively. Northern blot analysis indicated that AtHD2A was highly expressed in flowers and young siliques of Arabidopsis plants, whereas AtHD2B was widely expressed in stems, leaves, flowers and young siliques. AtHD2A repressed transcription when directed to a promoter containing GAL4-binding sites as a GAL4 fusion protein. Deletion of the extended acidic domain or the domain containing predicted catalytic residues of AtHD2A resulted in the loss of gene repression activity, revealing the importance of both domains to AtHD2A function. Arabidopsis plants were transformed with a gene construct comprising an AtHD2A cDNA in the antisense orientation driven by a strong constitutive promoter, 394tCUP. Silencing of AtHD2A expression resulted in aborted seed development in transgenic Arabidopsis plants, suggesting that the AtHD2A gene product was important in the reproductive development of Arabidopsis thaliana. PMID- 10792818 TI - Three ethylene-responsive transcription factors in tobacco with distinct transactivation functions. AB - Ethylene-responsive factors (ERFs) have conserved DNA-binding domains and interact directly with the GCC box in the ethylene-responsive element that is necessary and sufficient for the regulation of transcription by ethylene. ERFs were shown to be localized to nucleus in transient transfection experiments. Transient expression assays using tobacco protoplasts and a heterologous system in yeast were used to examine the transactivation functions of ERFs. ERF2 and ERF4 enhanced the GCC box-mediated transcription of a reporter gene in tobacco protoplasts. When fused to the DNA-binding domain of yeast GAL4, a carboxy terminal region of ERF2, as well as both amino-terminal and carboxy-terminal regions of ERF4, functioned as a transactivation domain in tobacco protoplasts. The amino-terminal regions of ERF2 and ERF4 functioned as transactivation domains in yeast. In contrast to ERF2 and ERF4, ERF3 reduced the transcription of the reporter gene in tobacco protoplasts, indicating that ERF3 functions as a repressor. Thus, it appears that ERFs exert their regulatory functions in different ways, with ERF2 and ERF4 being activators and ERF3 being a repressor of transcription. PMID- 10792819 TI - Understanding in vivo carbon precursor supply for fatty acid synthesis in leaf tissue. AB - The principal supply of carbon precursors for fatty acid synthesis in leaf tissue has been a much debated topic, with some experiments suggesting a direct supply from the C3 products of photosynthetic carbon fixation and colleagues suggesting the utilization of free acetate (for which concentrations in leaves in the range of 0.05-1.4 mM have been reported). To address this issue we first reassessed the in vivo rate of fatty acid synthesis using a new method, that of [13C]carbon dioxide labeling of intact Arabidopsis plants with the subsequent analysis of fatty acids by gas chromatography-mass spectrometry (GC-MS). This method gave an average value of 2.3 mmoles carbon atoms h-1 mg chlorophyll-1 for photosynthetic tissues. The method was extended by isotopic dilution analysis to measure the rate of fatty acid synthesis in the dark. There was negligible fatty acid synthesis (< 5% of the rate in the light) in the dark. In addition, the method allowed an estimate of the absolute rate of fatty acid degradation of about 4% of the total fatty acid content per day. With the in vivo rate of fatty acid synthesis in the light defined, if the bulk tissue acetate concentration available for fatty acid synthesis is 1 mM, this acetate pool can sustain fatty acid synthesis for approximately 60 min. When the leaves of Arabidopsis, barley and pea were given a 5 min pulse of [14C]carbon dioxide, the label rapidly appeared in fatty acids with a lag phase of less than 2-3 min. Continuous labeling with [14C]carbon dioxide, for up to 1 h, showed a similar result. Furthermore, 14C-label in free acetate was less than 5% of that in fatty acids. In conclusion, these data suggest that either the bulk pool of acetate is not involved in fatty acid synthesis or the concentration of acetate must be less than 0.05 mM under strong illumination. PMID- 10792820 TI - Plant viral suppressors of post-transcriptional silencing do not suppress transcriptional silencing. AB - Homology-dependent gene silencing is a regulatory mechanism that limits RNA accumulation from affected loci either by suppression of transcription (transcriptional gene silencing, TGS) or by activation of a sequence-specific RNA degradation process (post-transcriptional gene silencing, PTGS). The P1/HC-Pro sequence of plant potyviruses and the 2b gene of the cucumber mosaic virus have been shown to interfere with PTGS. The ability of these viral suppressors of PTGS to interfere with TGS was tested using the 271 locus which imposes TGS on transgenes under 35S or 19S promoters and PTGS on the endogenous nitrite reductase gene (Nii). Both P1/HC-Pro and 2b reversed PTGS of Nii genes in 271 containing tobacco plants, but failed to reverse TGS of 35S-GUS transgenes in the same plant. P1/HC-Pro expression from a transgene also failed to suppress either the initiation or maintenance of TGS imposed by the NOSpro-silencing locus, H2. These results indicate that PTGS and TGS operate through unlinked pathways or that P1/HC-Pro and 2b interfere at step(s) in PTGS that are downstream of any common components in the two pathways. The data suggest a simple assay to identify post-transcriptionally silenced transgenic lines with the potential to be stably converted to high expressing lines. PMID- 10792821 TI - Heterotrimeric G-proteins of a filamentous fungus regulate cell wall composition and susceptibility to a plant PR-5 protein. AB - Membrane permeabilizing plant defensive proteins first encounter the fungal cell wall that can harbor specific components that facilitate or prevent access to the plasma membrane. However, signal transduction pathways controlling cell wall composition in filamentous fungi are largely unknown. We report here that the deposition of cell wall constituents that block the action of osmotin (PR-5), an antifungal plant defense protein, against Aspergillus nidulans requires the activity of a heterotrimeric G-protein mediated signaling pathway. The guanidine nucleotide GDPbetaS, that locks G-proteins in a GDP-bound inactive form, inhibits osmotin-induced conidial lysis. A dominant interfering mutation in FadA, the alpha-subunit of a heterotrimeric G-protein, confers resistance to osmotin. A deletion mutation in SfaD, the beta-subunit of a heterotrimeric G-protein also increases osmotin resistance. Aspergillus nidulans strains bearing these mutations also have increased tolerance to SDS, reduced cell wall porosity and increased chitin content in the cell wall. PMID- 10792822 TI - A lipochito-oligosaccharide, Nod factor, induces transient calcium influx in soybean suspension-cultured cells. AB - Lipochito-oligosaccharides (Nod factors) produced by Rhizobium or Bradyrhizobium are the key signal molecules for eliciting nodulation in their corresponding host legumes. To elucidate the signal transduction events mediated by Nod factors, we investigated the effects of Nod factors on the cytosolic [Ca2+] of protoplasts prepared from roots and suspension-cultured cells of soybean (Glycine max and G. soja) using a fluorescent Ca2+ indicator, Fura-PE3. NodBj-V (C18:1, MeFuc), which is a major component of Nod factors produced by Bradyrhizobium japonicum, induces transient elevation of cytosolic [Ca2+] in the cells of soybean within a few minutes. This effect is specific to soybean cells and was not observed in the tobacco BY-2 cells. Furthermore, NodBj-V without MeFuc did not induce any cytosolic [Ca2+] elevation in soybean cells. Exclusion of Ca2+ from the medium, as well as pre-treatment of the cells with an external Ca2+ chelator or with a plasma membrane voltage-dependent Ca2+ channel inhibitor, suppressed the Nod factor-dependent cytosolic [Ca2+] elevation. These results indicate that transient Ca2+ influx from extracellular fluid is one of the earliest responses of soybean cells to NodBj-V (C18:1, MeFuc) in a host-specific manner. PMID- 10792823 TI - Heat-shock tagging: a simple method for expression and isolation of plant genome DNA flanked by T-DNA insertions. AB - This paper describes expression profiles of the Arabidopsis HSP18.2 heat-shock gene promoter by using three different reporter genes, and the application of this promoter to a method we have developed to drive heat-shock-dependent transcription of plant genome DNA flanked by T-DNA insertions. We show that, irrespective of the location of the T-DNA insertion, an HSP18.2 promoter towards the left border of the T-DNA effectively induces transcription of flanking genome sequences in response to heat shock. If polyadenylated, tagged transcripts can be easily identified by RT-PCR. PMID- 10792824 TI - Cis-elements and trans-factors that regulate expression of the maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2O2 is the likely intermediary signaling molecule for the response. AB - The mechanisms by which the maize antioxidant Cat1 gene responds to abscisic acid (ABA) and osmotic stress have been investigated. Results show that during late embryogenesis, Cat1 expression in vivo is independent of endogenous ABA levels. However, exogenously applied ABA significantly enhances Cat1 expression. Transient assays using particle bombardment show that the proximal ABRE2 element on the Cat1 promoter is responsible for the induction of Cat1 expression by ABA. We further show that ABA induces the expression of Cat1 via the interaction between ABRE2 and one of its binding proteins, CBF1 (Cat1 binding factor 1). Using ABA-deficient mutant embryos, we show that osmotic stress induces Cat1 expression through two alternate signal transduction pathways: an ABA signaling pathway leading to the interaction between the ABRE2 motif and CBF1, and a pathway via the interaction of ABRE2 and CBF2 (Cat1 binding factor 2) that is independent of ABA. The data presented clearly suggest that hydrogen peroxide (H2O2) plays an important intermediary role in the ABA signal transduction pathway leading to the induction of the Cat1 gene. PMID- 10792825 TI - The two largest chloroplast genome-encoded open reading frames of higher plants are essential genes. AB - The chloroplast genomes of most higher plants contain two giant open reading frames designated ycf1 and ycf2. In tobacco, ycf1 potentially specifies a protein of 1901 amino acids. The putative gene product of the ycf2 reading frame is a protein of 2280 amino acids. In an attempt to determine the functions of ycf1 and ycf2, we have constructed several mutant alleles for targeted disruption and/or deletion of these two reading frames. The mutant alleles were introduced into the tobacco plastid genome by biolistic chloroplast transformation to replace the corresponding wild-type alleles by homologous recombination. Chloroplast transformants were obtained for all constructs and tested for their homoplastomic state. We report here that all transformed lines remained heteroplastomic even after repeated cycles of regeneration under high selective pressure. A balanced selection was observed in the presence of the antibiotic spectinomycin, resulting in maintenance of a fairly constant ratio of wild-type versus transformed genome copies. Upon removal of the antibiotic and therewith release of the selective pressure, sorting out towards the wild-type plastid genome occurred in all transplastomic lines. These findings suggest that ycf1 and ycf2 are functional genes and encode products that are essential for cell survival. The two reading frames are thus the first higher plant chloroplast genes identified as being indispensable. PMID- 10792826 TI - Temporal and spatial regulation of pectic (1-->4)-beta-D-galactan in cell walls of developing pea cotyledons: implications for mechanical properties. AB - Modifications in cell wall pectic polysaccharides are thought to influence cell cell adhesion and the mechanical properties of plant tissues. Monoclonal antibodies to epitopes occurring in homo- galacturonan and side chains of rhamnogalacturonan I have been used in an immunolocalization study of cell wall architecture of developing pea cotyledons. Pectic (1-->4)-beta-D-galactan appears in cotyledon cell walls at a defined stage late in development (approximately 26 30 days after anthesis), whereas homogalacturonan and pectic (1-->5)-alpha-L arabinan are present in cotyledon cell walls throughout development. (1-->4)-beta galactan was restricted to a distinct thin layer at the plasma membrane face of the cell wall. Anion exchange and immunoaffinity chromatography indicated that the (1-->4)-beta-galactan was associated with acidic pectic components. Mechanical compressive testing of pea cotyledons, before and after (1-->4)-beta galactan appearance, indicated that the cotyledons with the galactan-rich cell wall layer were twice as firm as those with no detectable (1-->4)-beta-galactan. PMID- 10792827 TI - Disruption of the Arabidopsis photosystem I gene psaE1 affects photosynthesis and impairs growth. AB - The psae1-1 mutant of Arabidopsis was identified on the basis of a decrease in the effective quantum yield of photosystem II, among a collection of plants subjected to transposon tagging with the Enhancer element. The steady-state redox level and the rate of re-oxidation of P700 are significantly altered in psae1-1 mutants. The responsible mutation was localised to psaE1, one of two Arabidopsis genes that encode subunit E of photosystem I. An additional mutant allele, psae1 2, was identified by reverse genetics. In wild-type plants, the psaE1 transcript is expressed at a higher level than psaE2 mRNA. In the mutants, however, the E1 transcript was barely detectable, and was expressed only in small groups of wild type cells resulting from somatic reversions. As a consequence, the amount of PsaE protein present in the mutant is significantly reduced. Concomitantly, the levels of other stromal photosystem I subunits (PsaC and PsaD) are also affected. Mutant plants showed a marked increase in light sensitivity and photoinhibition. Additional effects of the psae1 mutation include light green pigmentation, an increase in chlorophyll fluorescence and a decrease of approximately 50% in growth rate under greenhouse conditions. PMID- 10792828 TI - Light-induced nuclear import of phytochrome-A:GFP fusion proteins is differentially regulated in transgenic tobacco and Arabidopsis. AB - Phytochromes (phy) are a family of photoreceptors that control various aspects of light-dependent plant development. Phytochrome A (phyA) is responsible for the very low fluence response (VLFR) under inductive light conditions and for the high irradiance response (HIR) under continuous far-red light. We have recently shown that nuclear import of rice phyA:GFP is regulated by VLFR in transgenic tobacco. The import is preceded by very fast, light-induced formation of sequestered areas of phyA:GFP in the cytosol. Here we report that expression of the Arabidopsis phyA:GFP fusion protein in phyA-deficient Arabidopsis plants complements the mutant phenotype. In these transgenic Arabidopsis lines, both light-dependent cytosolic formation of sequestered areas of the phyA:GFP as well as VLFR or HIR-mediated nuclear import of the fusion protein was observed. By contrast, light-dependent nuclear import of the same fusion protein was induced only by continuous far-red light (HIR) but not by pulses of far-red light (VLFR) in transgenic tobacco. These results demonstrate that photoregulation of intracellular partitioning of the Arabidopsis phyA:GFP differs significantly in different genetic backgrounds. PMID- 10792829 TI - Photocontrol of subcellular partitioning of phytochrome-B:GFP fusion protein in tobacco seedlings. AB - Photomorphogenesis of higher plants is regulated by photoreceptors including the red/far-red light-absorbing phytochromes, blue-UV/A sensing cryptochromes and as yet uncharacterized UV/B receptors. Specific phototransduction pathways that are controlled by either individual or interacting photoreceptors mediate regulation. Phytochrome B (phyB) is the major red light-sensing photoreceptor. Phototransduction mediated by this light sensor has been shown to include light dependent nuclear import and interaction of phyB with transcription factor-like proteins in the nucleus. Here we report that nuclear import of phyB and physiological responses regulated by this photoreceptor exhibit very similar wavelength- and fluence rate-dependence. Nuclear import of phyB is insensitive to single red, blue and far-red light pulses. It is induced by continuous red light and to a lesser extent by continuous blue light, whereas far-red light is completely ineffective. The data presented indicate that light-dependent partitioning of phyB exhibits features characteristic of blue light responsiveness amplification, a phenomenon that is thought to be mediated by interaction of phyB with CRY1. PMID- 10792830 TI - Hyperosmotic stress stimulates phospholipase D activity and elevates the levels of phosphatidic acid and diacylglycerol pyrophosphate. AB - In mammalian cells, phospholipase D (PLD) and its product phosphatidic acid (PA) are involved in a number of signalling cascades, including cell proliferation, membrane trafficking and defence responses. In plant cells a signalling role for PLD and PA is also emerging. Plants have the extra ability to phosphorylate PA to produce diacylglycerol pyrophosphate (DGPP), a newly discovered phospholipid whose formation attenuates PA levels, but which could itself be a second messenger. Here we report that increases in PA and its conversion to DGPP are common stress responses to water deficit. Increases occur within minutes of treatment and are dependent on the level of stress. Part of the PA produced is due to PLD activity as measured by the in vivo transphosphatidylation of 1 butanol, and part is due to diacylglycerol kinase activity as monitored via 32P PA formation in a differential labelling protocol. Increases in PA and DGPP are found not only in the green alga Chlamydomonas moewusii and cell-suspension cultures of tomato and alfalfa when subjected to hyperosmotic stress, but also in dehydrated leaves of the resurrection plant Craterostigma plantagineum. These results provide further evidence that PLD and PA play a role in plant signalling, and provide the first demonstration that DGPP is formed during physiological conditions that evoke PA synthesis. PMID- 10792831 TI - Decreased and increased expression of the subunit CHL I diminishes Mg chelatase activity and reduces chlorophyll synthesis in transgenic tobacco plants. AB - The chelation of Fe2+ and Mg2+ ions forms protoheme IX and Mg-protoporphyrin IX, respectively, and the latter is an intermediate in chlorophyll synthesis. Active magnesium protoporphyrin IX chelatase (Mg-chelatase) is an enzyme complex consisting of three different subunits. To investigate the function of the CHL I subunit of Mg-chelatase and the effects of modified Mg-chelatase activity on the tetrapyrrole biosynthetic pathway, we characterized N. tabacum transformants carrying gene constructs with the Chl I cDNA sequence in antisense and sense orientation under the control of the CaMV 35S promoter. Both elevated and diminished levels of Chl I mRNA and Chl I protein led to reduced Mg-chelatase activities, reflecting a perturbation of the assembly of the enzyme complex. The transformed plants did not accumulate the substrate of Mg-chelatase, protoporphyrin IX, but the leaves contained less chlorophyll and possessed increased chlorophyll a/b ratios, as well as a deficiency of light-harvesting chlorophyll binding proteins of photosystems I and II. The expression and activity of several tetrapyrrolic enzymes were reduced in parallel to lower the Mg-chelatase activity. Consistent with the lower chlorophyll contents, the rate limiting synthesis of 5-aminolevulinate was also decreased in the transgenic lines analyzed. The consequence of reduced Mg-chelatase on early and late steps of chlorophyll synthesis, and on the organization of light harvesting complexes is discussed. PMID- 10792832 TI - A pollen tube growth-promoting arabinogalactan protein from nicotiana alata is similar to the tobacco TTS protein. AB - Upon germination on the stigma, pollen tubes elongate in the stylar transmitting tract, aided by female factors, with speed and directionality not mimicked in in vitro pollen tube growth cultures. We have shown that a stylar transmitting tissue arabinogalactan protein (AGP) from Nicotiana tabacum (tobacco), TTS protein, stimulates pollen tube growth in vivo and in vitro and attracts pollen tubes grown in a semi-in vivo culture system. It has been reported that the self incompatible Nicotiana alata produced a stylar glycoprotein, GaRSGP, which had a backbone polypeptide that shared 97% identity with those of TTS proteins but some of its properties were different from those described for TTS proteins. We report here the characterization of a family of stylar transmitting tissue glycoproteins from N. alata that is virtually identical to tobacco TTS proteins and which we refer to as NaTTS proteins. Like their tobacco counterparts, NaTTS proteins are recognized by the traditional AGP-diagnostic reagent beta-glucosyl Yariv reagent, and they are also recognized by JIM13, a monoclonal antibody against AGP. NaTTS proteins also stimulate pollen tube elongation in vitro and attract pollen tubes in a semi-in vivo pollen tube culture system. Biochemical and immunological characterization of NaTTS proteins revealed that they have extraordinary variability in the extent of sugar modifications of their polypeptide backbones. The extent of sugar modifications on NaTTS proteins significantly affects their biochemical properties, influences how they interact with the transmitting tissue extracellular matrix, and affects their solubility from this matrix. Our results suggest that the strategy used to purify GaRSGP only recovered a less glycosylated, more tightly extracellular matrix-bound sub-population of the entire spectrum of N. alata TTS proteins. PMID- 10792833 TI - Chemokine expression during acute rejection of rat kidneys. AB - During acute rejection of fully allogeneic rat renal allografts, few neutrophil granulocytes are detected, whereas an abundant infiltrate of macrophages and T lymphocytes becomes apparent. The mechanisms leading to this specific pattern of infiltration are not understood. We performed a sequential daily Northern blot analysis of the mRNA expression of the CC-chemokines MCP-1, MIP-1alpha and RANTES and of the CXC-chemokines GRO/KC and MIP-2 in rat renal isografts (LEW --> LEW, n = 1 per day) and allografts during acute rejection (DA --> LEW, n = 3 per day). MCP-1 gene expression strongly increased on days 3-4 after allotransplantation and returned to control levels on day 6. The expression of MIP-1alpha and RANTES continuously rose until day 3-4 and remained stable thereafter. Isografts displayed minor changes in CC-chemokine expression. In contrast to CC-chemokines, GRO/KC was expressed in low amounts during rejection and MIP-2 mRNA remained undetectable. In conclusion, the expression of the CC-chemokines MCP-1, MIP-1 and RANTES was clearly upregulated during rejection, whereas the mRNA of the CXC chemokines MIP-2 and GRO/KC was not detected at all or remained at low levels. This pattern of chemokine gene expression is in good accordance with the predominant mononuclear leukocyte infiltrate in allografts. PMID- 10792834 TI - Anti-CD3 induced thymocyte apoptosis in vivo require the antibody Fc domain. AB - We have earlier found that explanted thymic epithelial cells (TEC) can produce glucocorticoid (GC) activity in vitro and that the GC receptor (GR) antagonist RU486 partially inhibit thymic apoptosis induced by the anti-CD3 monoclonal antibodies (MoAb) 2C11, both in vivo and in new-born thymic organ cultures. To explain the inhibitory effect of RU486 in this system we have now investigated the importance of the 2C11 Fc as this MoAb bind with high affinity to cellular FcR. We have found both that whole 2C11 MoAb can bind to explanted TEC in vitro and that F(ab)'2 fragments from this MoAb loose this ability, in addition with the capacity to induce thymic apoptosis in vivo. We interpret our results to indicate that the injected 2C11 MoAb may establish a close contact between GC producing, FcR positive TEC cells and CD3 positive thymocytes and thereby subject the later to high paracrine GC concentrations and subsequent induction of apoptosis. PMID- 10792835 TI - Autoantibodies to cryptic epitopes elicited by infection with lactate dehydrogenase-elevating virus. AB - Lactate dehydrogenase-elevating virus (LDV) produces a permanent infection in mice with a B-lymphocyte polyclonal activation leading to hypergammaglobulinaemia. Since LDV specifically suppressed antibodies to native epitopes in CBA/Ht, but not BALB/c, mice immunized against a protein antigen, we explored the relationship between such a change in antibody specificity and the expression of autoantibodies under the influence of LDV. Again in CBA/Ht, but not BALB/c, mice we observed another effect of LDV: the sera from infected CBA/Ht mice were found by enzyme-linked immunosorbant assay to contain antibodies to various mouse tissue extracts. Immunoblots revealed a large spectrum of autoantigens that differed markedly between animals. Western-blot competition experiments showed that the protein autoantigens had to be denatured to react with most of the autoantibodies. Despite the presence of these autoantibodies directed to cryptic epitopes, no specific tissue lesions could be ascribed to the autoimmune response elicited by LDV infection, since both mouse strains showed mild inflammatory reactions in liver and kidney. PMID- 10792836 TI - B-cell depletion fails to abrogate the induction of oral tolerance of specific Th1 immune responses in mice. AB - Antigen presentation by resting B cells has been shown to induce peripheral tolerance to intravenous (i.v.) administered soluble protein antigens. We further examined the role of resting B cells in the induction of oral tolerance. Mice were treated continuously from birth with rabbit antimouse IgM serum for 5 weeks. Immunohistological studies revealed that anti-IgM treatment depleted B cell aggregated follicles in intestinal Peyer's patches. At 4-weeks-old, B cell depleted mice were fed 25 mg ovalbumin or given 10% chicken egg white to drink for 5 days. Anti-IgM treatment was stopped 2 days after the last feed. Ten weeks later, the mice were immunized with 100 microg ovalbumin emulsified with complete Frund's adjuvant. Their T helper 1 (Th1) cell-regulated systemic delayed-type hypersensitivity, IgG2a antibody responses and spleen cell production of interferon-r and interleukin-2 were suppressed by prior ovalbumin or egg white feeding during anti-IgM treatment. Active suppression of Th1 immune responses was also demonstrated following adoptive transfer of egg white-fed donor spleen cells collected during anti-IgM treatment to naive recipients. Although enormous small resting B cells are aggregated in the mantle zones of follicles of intestinal Peyer's patches, they are not the antigen-presenting cells seen in the induction of oral tolerance. PMID- 10792837 TI - The role of interleukin-4 in the regulation of sequential isotype switch from immunoglobulin G1 to immunoglobulin E antibody production. AB - The immunoglobulin (Ig)E immune response against protein antigens is profoundly influenced by the antigen dose used for immunization. Whereas immunization of CBA/J mice with low antigen doses results in the production of large amounts of IgE antibody, priming with high antigen doses leads to only marginal IgE antibody production in animals. However, in vitro restimulation of spleen cells from mice primed with high antigen doses leads to considerable activation of IgE-producing B cells, which suggests that B cells primed for IgE antibody production do exist among spleen cells. We investigated the modalities of activation of these memory B cells. The data presented here reveal that the anamnestic IgE immune response in vitro is strictly dependent on the presence of IgG1-expressing B cells, which differentiate after a sequential isotype switch into IgE-producing plasma cells with the help of primed CD4+ T cells. The induction of IgE-producing plasma cells requires a cognate interaction between B cells and CD4+ T cells. Interleukin (IL) 4 seems not to be involved in this process, since IgE production in vitro is resistant to suppression by anti-IL-4 monoclonal antibody. Finally, we show that IgG1-expressing B cells represent a relevant memory cell population in vivo also, but in contrast to the in vitro situation the final differentiation into IgE producing plasma cells is dependent on IL-4. PMID- 10792838 TI - The early immune response in the liver of BALB/c mice infected with S. typhimurium. AB - Gram-negative bacteria acquired through gastrointestinal infection can be a serious cause for the development of septic shock especially in immunosuppressed patients. Thus, the aim of this study was to examine the early events of the immune reaction against S. typhimurium. Bacteria were injected into mice at different concentrations. Four animals from each group were killed at five different points of time. Liver cytokine mRNA expression was determined by semiquantitative rt-PCR and liver histology was examined. Serum cytokine levels of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-4 and IL-10 were determined. intravenous (i.v.) infection with 109 bacteria led to lethal septic shock within 24 h. A delayed production of IFN-gamma, TNF alpha, IL-18 and IL-10 and milder histological alterations in the liver were observed in these animals. The highest expression of cytokines in the liver and the strongest histological alterations were seen after infection with 107 bacteria. Here, an increased mRNA expression of all proinflammatory cytokines began 1 h after infection. Animals infected with 1 x 102 bacteria had the highest detectable serum levels of IL-6 and IL-10. These data indicate that the immediate events in the immune reaction within the liver after infection with S. typhimurium are associated with the outcome of the subsequent sepsis. PMID- 10792839 TI - Heat-aggregated immunoglobulins increase in vivo immunogenicity of mouse hapten (TNP)-derivatized macrophages by upregulation of interleukin-12 secretion and expression of B7-1 and B7-2 costimulatory molecules. AB - Antigen-antibody complexes (IC) can up or down regulate immune responses by induction of immunoregulatory cells. We have studied the effect of mouse heat aggregated immunoglobulin (Ig) (HA) which have many biological activities similar to IC on immunogenicity of TNP-substituted macrophages (TNP-Mphi). Our results show that: (1) mouse oil-induced peritoneal macrophages treated with HA produce in vitro significantly higher levels of interleukin (IL-1beta), tumor necrosis factor (TNF)-alpha, IL-6, IL-10 and particularly IL-12 and express more B7-1 and B7-2 and ICAM-1 cell surface costimulatory molecules than Mphi treated with monomeric Ig (MM); (2) Mphi derivatized with TNP, treated or not with MM, induce in vivo antigen-specific unresponsiveness. In contrast TNP-Mphi treated with HA induce significant contact sensitivity reaction even when injected into previously tolerized recipient animals. Treatment of recipients with anti-IL-12 Ab prevents immunization by TNP-Mphi-HA. These results indicate that bypass of tolerance by treatment of TNP-Mphi with HA is a result of an increased production of IL-12 by these cells and an enhanced expression of costimulatory molecules important in T cell-Mphi interactions. We suggest that a similar overcoming of tolerance through the action of IC may be responsible for the generation of autoantibodies of heterologous specificity in pathological conditions in which such complexes are formed. PMID- 10792840 TI - Modulation of neutrophil influx with cell adhesion molecule specific antibodies during nonspecific and immune mediated inflammatory reactions. AB - Neutrophils are essential for the host defence against infection. However, neutrophils may also mediate damage namely during immune mediated pathologies. We therefore tested whether targeting of different cell adhesion molecules with specific monoclonal antibodies might reduce immune mediated neutrophil recruitment but spare the nonspecific accumulation of neutrophils that is essential for the resistance against acute infections. Neutrophil recruitment was induced by either intraperitoneal injection of casein as a nonspecific phlogistic agent or by i.p. injection of antigen in Mycobacterium bovis Bacille Calmette Guerin (BCG) immune mice. Similar degrees of inhibition of neutrophil accumulation were observed in both models of inflammation with antibodies directed at CD11a, ICAM-1 and CD11b with the latter showing the most marked effects. Individual targeting of selectins was without effect in immune mediated responses whereas targeting of L or E selectin inhibited nonspecific recruitment of neutrophils. This was apparently not owing to a dosage effect nor to a kinetic difference. The inhibitory effect of anti-CD11b antibodies was most likely as a result of activation of circulating neutrophils rather than the blocking of receptor-ligand interactions. We were therefore unable to selectively abrogate immune mediated neutrophil recruitment with the use of the antibodies selected in this study. PMID- 10792841 TI - The phosphatase domains of CD45 are required for ligand induced T-cell receptor downregulation. AB - Down-regulation of the T-cell receptor (TCR) plays an important role in modulating T-cell responses, both during T-cell development and in mature T cells. At least two distinct pathways exist for TCR down-regulation: down regulation following TCR ligation; and down-regulation following activation of protein kinase C (PKC). Ligand-induced TCR down-regulation is dependent on protein tyrosine kinase (PTK) activity and seems to be closely related to T-cell activation. In addition, previous studies have indicated that ligand-induced TCR down-regulation is dependent on the expression of CD45, a transmembrane protein tyrosine phosphatase. The role of the different domains of CD45 in TCR down regulation was investigated in this study. We found that the phosphatase domains of CD45 are required for efficient ligand-induced TCR down-regulation. In contrast, the extracellular domain of CD45 is dispensable for ligand-mediated TCR down-regulation. Finally, PKC-mediated TCR down-regulation was found to be independent of both the extra-and intracellular domains of CD45. PMID- 10792842 TI - Multiepitope vaccines intensively increased levels of antibodies recognizing three neutralizing epitopes on human immunodeficiency virus-1 envelope protein. AB - A few neutralizing antibodies against human immunodeficiency virus-1 (HIV-1) envelope proteins have been shown to be highly effective at neutralizing different strains in vitro, and exist at very low levels in the sera of HIV-1 infected individuals. Based on our hypothesis that epitope vaccination may be a novel strategy for inducing high levels of antibodies against HIV-1, we prepared multiepitope vaccines using three neutralizing epitopes (GPGRAFY, ELDKWA and RILAVERYLKD) on HIV-1 envelope proteins. The PI [C-G-(ELDKWA-GPGRAFY)2-K] and PII (CG-GPGRAFY-G-ELDKWA-G-RILAVERYLKD) peptides were synthesized and conjugated to a carrier protein, bovine serum albumin (BSA). After vaccination, both the PI-BSA and PII-BSA multiepitope vaccines induced high levels of epitope-specific antibodies to the three neutralizing epitopes (antibody titre: 1 : 12,800 102,400). The recombinant glycoprotein 160 (rgp160) subunit vaccine induced strong antibody responses to rgp160, but only very weak epitope-specific antibody responses to the three epitopes. The epitope-specific antibodies were isolated from rabbit sera by single epitope-peptide-conjugated sepharose columns. A yield of 51 microg of epitope-specific antibodies/ml of serum (mean value) was obtained and identified to recognize these epitopes, while 0.35 microg of protein was isolated from 1 ml of pooled preserum by C-(ELDKWAG)4- or C-(RILAVERYLKD-G)2-K- and C-(GPGRAFY)4-sepharose columns. The levels of these epitope-specific antibodies induced in rabbits were much greater than 1 microg/ml, a level that is considered to confer long-term protection against some viruses. Moreover, these antibodies recognized the neutralizing epitopes on peptides and rgp41. Based on the fact that a very low level of ELDKWA epitope-specific antibodies exist in HIV 1-infected individuals, these results suggesting that synthetic epitope vaccines could induce high levels of multiepitope-specific neutralizing antibodies indicate a new strategy for developing an effective neutralizing antibody-based epitope/peptide vaccine against HIV-1. PMID- 10792843 TI - Mechanisms involved in protective immune response generated by secretory proteins of Mycobacterium habana against experimental tuberculosis. AB - Live mycobacteria secrete a number of unique proteins early in their multiplication which are important for both the pathogenesis and the stimulation of specific host responses. We have investigated the mechanisms by which the host mounts immune response against tuberculosis after vaccination with secretory proteins (SP) of a vaccine candidate Mycobacterium habana TMC 5135. Mice vaccinated with SP of 10th day growth of M. habana, either alone or emulsified in Freund's incomplete adjuvant (FIA) possessed antituberculous resistance and cellular immune responses against M. tuberculosis H37Rv. These proteins induced a significant cutaneous delayed type hypersensitivity response in guinea pigs vaccinated with heat killed M. tuberculosis H37Rv, which was equivalent to that observed with a standard purified protein derivative (PPD). The splenocytes of these guinea pigs have shown higher proliferative response after stimulation with SP than with PPD. The SP + FIA immunization has been found to exert maximum prophylactic effect by potentiating both the oxygen dependent arms and enzymatic activities of macrophages. Macrophages from mice vaccinated with SP of M. habana produced enhanced levels of interleukin(IL)-2, interleukin-12 and interferon(IFN) gamma. The protective as well as cell mediated immune responses were upregulated in SP immunized animals when compared to whole cell (M. habana) vaccinated animals. SDS-PAGE of SP from M. habana showed the prominent bands of 60, 32, 31 and 30 kDa. Furthermore, the western analysis of SP with pulmonary tuberculosis patient's serum has revealed the presence of immunoreactive antigens of 36, 35, 33/32 kDa. Overall study demonstrated that the secretory antigens released by actively growing M. habana bacilli could activate different arms of effective immune response. PMID- 10792845 TI - Immunological and virological changes in antiretroviral naive human immunodeficiency virus infected patients randomized to G-CSF or placebo simultaneously with initiation of HAART. AB - To determine the efficacy of combined G-CSF and highly active antiretroviral treatment (HAART), a randomized, double blind, placebo controlled study was conducted. Treatment naive human immunodeficiency virus (HIV) infected patients were randomized to receive either placebo or G-CSF (0.3 mg/ml, 3 times a week) for 12 weeks and HAART simultaneously. The trial was terminated prematurely after interim analysis performed because of a case of severe encephalopathia in the G CSF group. At that point 11 HIV infected patients with a CD4+ T cell count < 350/mm3 had been randomized to the G-CSF group (n = 6) or placebo group (n = 5). In both groups plasma HIV RNA decreased significantly in response to HAART. However, plasma HIV RNA changed significantly different between the two groups with the decrease being less pronounced in the G-CSF group (P = 0.02). The concentrations of CD4+ memory T cells and CD8+ naive and memory T cells increased in response to HAART, and there was a trend towards more pronounced increases in several T-cell subpopulations in the G-CSF group. The CD56+ NK cells increased significantly more in the G-CSF group compared with placebo (P = 0. 000). All patients in the G-CSF group reported bone pain. The present data do not support simultaneous administration of G-CSF with initiation of HAART in treatment naive HIV infected patients. PMID- 10792844 TI - T-cell repertoire analysis in acute and chronic human Chagas' disease: differential frequencies of Vbeta5 expressing T cells. AB - Here, we analysed the use of Vbeta-TCR regions by CD4+ and CD8+ T cells from acute and chronic chagasic patients using flow cytometry. We determined the Vbeta expression in cells freshly isolated from patients, as well as after in vitro stimulation with antigens derived from epimastigote (EPI) or trypomastigote (TRYPO) forms of Trypanosoma cruzi. Analysis of Vbeta-TCR expression of T cells freshly isolated from patients showed a decrease in Vbeta5 expression in the CD4+ T-cell population from acutely infected individuals, whereas CD4+Vbeta5+ T cells were found to be increased in chronic patients with the cardiac, but not indeterminate, clinical form. After culturing peripheral blood mononuclear cells (PBMC) from chronic patients with EPI or TRYPO, we found that both antigenic preparations led to a preferential expansion of CD4+Vbeta5+ T cells. EPI stimulation also led to the expansion of CD8+Vbeta5+ T cells, whereas TRYPO led to the expansion of this cell population only if PBMC were from cardiac and not indeterminate patients. We observed that TRYPO stimulation led to an increase in the frequency of CD4+Vbeta17+ T cells in cultures of PBMC from indeterminate patients, whereas an increase in the frequency of CD8+Vbeta17+ T cells was found upon TRYPO stimulation of PBMC from cardiac patients. Despite this increase in the frequency of Vbeta17+ T-cell populations upon TRYPO stimulation, the same antigenic preparation led to a much higher expansion of Vbeta5+ T cells. These results show a differential expression of Vbeta5-TCR in cells freshly isolated from chagasic patients in different stages of the disease and that parasite specific antigens stimulate a portion of the T-cell repertoire with preferential usage of Vbeta5-TCR. PMID- 10792846 TI - The enhancing role of complement in human immunodeficiency virus infection: soluble recombinant CR1 (CD35) inhibits complement-mediated enhancement of infection of a CD4-positive T-cell line with human immunodeficiency virus-1. AB - The present study investigated the effect of soluble recombinant CR1 (srCR1, sCD35) on complement-dependent enhancement of human immunodeficiency virus-1 (HIV 1) infection in vitro. Cells of the human T-cell line HPB-ALL were infected with HIV-1 that had been preopsonized with normal human HIV-seronegative serum in the presence of srCR1. At nanomolar concentrations, srCR1 suppressed complement dependent enhancement of infection of HPB-ALL cells in a dose-dependent manner. Under these conditions, infection was decreased to levels similar to those observed in cells infected with unopsonized virus. These observations provide further evidence to support the role of complement-dependent opsonization facilitating viral entry into target cells. PMID- 10792847 TI - Binding of murine antibodies against whole-cell pertussis vaccine or filamentous haemagglutinin by Bordetella pertussis from patients with whooping cough. AB - In 1996 an unexpected rise in the incidence of whooping cough occurred in the Netherlands, and antigenic divergence between vaccine strains and clinical isolates has been suggested as a cause for this phenomenon. To investigate this assumption, the binding of murine antibodies against the whole-cell pertussis vaccine or filamentous haemagglutinin, an important protective antigen, to a limited number of Bordetella pertussis strains isolated during different time periods (1991-92, 1994 and 1996) was assessed. The results showed that all strains were recognized equally well by these antisera, indicating that filamentous haemagglutinin was unchanged during the time-periods examined. Although over the years changes have occurred in at least two surface proteins of B. pertussis, these changes are too subtle to be recognized by the antibodies raised in mice. Further research is required to assess whether antigenic variation of B. pertussis has an effect on protective immunity. PMID- 10792849 TI - A clinician's guide to the use of gonadotropin-releasing hormone analogues in women. AB - Gonadotropin-releasing hormone (GnRH) and its analogues have been extensively used in clinical medicine since they were identified and synthesized in 1971. Native GnRH stimulates gonadotrophs of the anterior pituitary and has been used for induction of ovulation. The GnRH agonists, which have greater potency and a longer half-life than native GnRH, produce an initial stimulation of pituitary gonadotrophs that results in secretion of follicle-stimulating hormone and luteinizing hormone and the expected gonadal response. This response is followed by downregulation and inhibition of the pituitary-gonadal axis. The GnRH antagonists promptly suppress pituitary gonadotropin by GnRH-receptor competition, thereby avoiding the initial stimulatory phase of the agonists. Discontinuation of GnRH antagonist treatment leads to a rapid and predictable recovery of the pituitary-gonadal axis. The GnRH analogues are potent therapeutic agents that are considerably useful in a variety of clinical indications. These indications include management of endometriosis, uterine leiomyomas, hirsutism, dysfunctional uterine bleeding, premenstrual syndrome, assisted reproduction, and some hormone-dependent tumors. PMID- 10792850 TI - Evaluating and managing premenstrual syndrome. AB - Premenstrual syndrome (PMS), a common disorder in women, refers to physical and/or mood symptoms that appear predictably during the latter half of the menstrual cycle, last until menses begin, and are absent during the early part of the menstrual cycle. A diagnosis of PMS requires that the symptoms be severe enough to affect a woman's ability to function at home or in the workplace or in her relationships with others. Diagnostic assessment entails a thorough medical and psychiatric history and prospective daily ratings. Disorders such as major depression, anxiety, hypothyroidism, and diabetes must be excluded before a diagnosis of PMS can be considered. Treatment strategies include either eliminating the hormonal cycle associated with ovulation or treating the symptom(s) causing the most distress to the patient. Medical therapies are available for both treatment approaches but should be initiated only after behavioral measures have failed; the physician must also carefully weigh the severity of symptoms against the potential for adverse effects of treatment. PMID- 10792848 TI - Distinct innate and acquired immune responses to Leishmania in putative susceptible and resistant human populations endemically exposed to L. (Viannia) panamensis infection. AB - Mechanisms of constitutive and acquired susceptibility/resistance to Leishmania Viannia panamensis (L. (V ) p.) were investigated in endemically exposed human populations presenting either recurrent disease (putative susceptible) or subclinical infection (clinically resistant). Cutaneous delayed type hypersensitivity response to leishmanin was significantly lower among individuals experiencing recurrent leishmaniasis than among those whose skin test converted without developing the disease. Monocyte derived macrophages from individuals with recurrent disease were more permissive in vitro to the entry of parasites than macrophages from subclinically infected individuals. In vitro proliferation of CD4 and CD8 T lymphocytes in response to intracellular amastigotes was significantly lower among individuals with a history of recurrent disease compared with subclinically infected individuals. Linear regression analyses revealed a strong direct relationship between the production of interferon (IFN) gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-10 by peripheral blood mononuclear cells (PBMC) from resistant (subclinically infected) individuals and no correlation in the production of these cytokines by PBMC from individuals who experienced recurrent disease. The results provide evidence of differences in the innate and acquired responses to Leishmania according to the outcome of the natural infection. These findings support the feasibility of identifying the immunological bases of innate and acquired resistance through studies in naturally exposed human populations. PMID- 10792851 TI - Women and schizophrenia. AB - Several important questions emerge from the study of gender differences in schizophrenia: Why does schizophrenia begin later in women? Why is outcome superior in women, at least in the first 15 years after onset? What causes sex differences in symptoms? What can gender differences teach us about the etiology of schizophrenia? Do men and women require substantially different treatments? What interventions during pregnancy and after childbirth ensure optimal health for the children of mothers with schizophrenia? Although complete answers may not yet be forthcoming, it is important to define the questions and keep them in mind when delivering services to women suffering from this severe, persistent mental illness. PMID- 10792854 TI - The agenda PMID- 10792853 TI - Osteoprotegerin and its ligand: A new paradigm for regulation of osteoclastogenesis and bone resorption. AB - In just 3 years, striking new advances have been made in understanding the molecular mechanisms that govern the crosstalk between osteoblasts/stromal cells and hemopoietic osteoclast precursor cells that leads to osteoclastogenesis. Led first by the discovery of osteoprotegerin (OPG), a naturally occurring protein with potent osteoclastogenesis inhibitory activity, rapid progress was made to the isolation of RANKL, a transmembrane ligand expressed on osteoblasts/stromal cells, that binds to RANK, a transmembrane receptor on hemopoietic osteoclast precursor cells. The interaction of RANK and RANKL initiates a signaling and gene expression cascade that results in differentiation and maturation of osteoclast precursor cells to active osteoclasts capable of resorbing bone. Osteoprotegerin acts as a decoy receptor; it binds to RANKL and blocks its interaction with RANK, thus inhibiting osteoclast development. Many of the calciotropic hormones and cytokines, including vitamin D3, parathyroid hormone, prostaglandin E2 and interleukin-11, appear to stimulate osteoclastogenesis through the dual action of inhibiting production of OPG and stimulating production of RANKL. Estrogen, on the other hand, appears to inhibit production of RANKL and RANKL-stimulated osteoclastogenesis. Recently, the results of the first clinical trial with OPG supported its potential as a therapeutic agent for osteoporosis. The new understanding provided by the RANK/RANKL/OPG paradigm for both differentiation and activation of osteoclasts has had tremendous impact on the field of bone biology and has opened new avenues for development of possible treatments of diseases characterized by excessive bone resorption. PMID- 10792855 TI - An analysis of the relationship between the utilization of physical therapy services and outcomes of care for patients after total hip arthroplasty. AB - BACKGROUND AND PURPOSE: The effect of physical therapy intervention on the outcomes of care for patients treated in acute care hospitals has not been widely studied. This study examined the relationship between physical therapy utilization and outcomes of care for patients following total hip arthroplasty. SUBJECTS: The sample consisted of 7,495 patients treated in US academic health center hospitals in 1996 who survived their inpatient stay and received physical therapy interventions. METHODS: The primary data source was the University HealthSystem Consortium Clinical Data Base. Physical therapy use was assessed by examining physical therapy charges. Outcomes of care were assessed in terms of the total cost of care (ie, whether the care was more costly or less costly than expected, taking into account patient characteristics) and in terms of discharge destination (ie, whether the patient was discharged home or elsewhere). Regression analyses were conducted to examine the relationship between physical therapy use and outcomes. RESULTS: Physical therapy intervention was directly related to a total cost of care that was less than expected and to an increased probability of discharge home. CONCLUSION AND DISCUSSION: The results of this study provide preliminary evidence to support the use of physical therapy intervention in the acute care of patients following total hip arthroplasty and indicate the need for further study of this topic. PMID- 10792856 TI - Experiences of older women with cancer receiving hospice care: significance for physical therapy. AB - BACKGROUND AND PURPOSE: The number of older adults with cancer is growing, increasing the need for professionals who are able to meet these patients' special needs. In palliative care settings, physical therapists strive to promote quality of life. Minimal research exists, however, to guide therapists working with patients with terminal illness. The purpose of this study was to gain knowledge that can be used by physical therapists to more effectively assess and treat older people with cancer receiving hospice care. SUBJECTS AND METHODS: A qualitative single-case study with replication was conducted with 3 older women with cancer who were receiving hospice care. Interview data were analyzed using grounded theory techniques. RESULTS: Four themes emerged as central to the experience of the informants: social relationships, spirituality, outlook on mortality, and meaningful physical activity. CONCLUSION AND DISCUSSION: In addition to maintaining physical function, physical therapists, who attend to nonphysical as well as physical aspects of care, may foster social cohesion, help maximize life's meaning, and support stabilizing strategies of older women with cancer who receive hospice care. PMID- 10792857 TI - Effectiveness of wound care products in the transmission of acoustic energy. AB - BACKGROUND AND PURPOSE: Ultrasound is often recommended in the treatment of people with partial and full-thickness wounds. Many treatments are performed over a hydrogel sheet or semipermeable film dressing. The purpose of this in vitro study was to examine the effectiveness of 4 hydrogels (Nu-Gel, ClearSite, Aquasorb Border, and CarraDres) and 4 film dressings (CarraSmart Film, J&J Bioclusive, Tegaderm, and Opsite Flexigrid) in ultrasound transmission. METHODS: The amount of sound energy transmitted through each product and interposed pig tissue was measured using an oscilloscope to display the intensity of sound energy delivered by the transducer. Five intensities at a frequency of 3.3 MHz were studied. RESULTS: Results were expressed as the mean ((SD) percentage of voltage transmitted compared with a gel baseline. Nu-Gel was the most efficient hydrogel (77.2%(4.6%), followed by ClearSite (72.0%(2.2%), Aquasorb Border (45.3%(2.1%), and CarraDres (42.8%(5.9%). The 4 film dressings, in order of efficiency, were CarraSmart Film (60.5%(4.4%), J&J Bioclusive (53.2%(2.4%), Tegaderm (47.1%(2.3%), and Opsite Flexigrid (31.5%(4.0%). CONCLUSION AND DISCUSSION: Transmissivity of wound care products used to deliver acoustic energy during ultrasound treatment of wounds varies greatly among dressing products. We believe that clinicians can use our findings as a part of the clinical reasoning process that they use to select an optimal wound dressing. PMID- 10792858 TI - Spinal cord control of movement: implications for locomotor rehabilitation following spinal cord injury. AB - In recent years, our understanding of the spinal cord's role in movement control has been greatly advanced. Research suggests that body weight support (BWS) walking and functional electrical stimulation (FES), techniques that are used by physical therapists, have potential to improve walking function in individuals with spinal cord injury (SCI), perhaps long after the stage of spontaneous recovery. Walking is one of the most desired goals of people with SCI; however, we are obligated to be judicious in our claims of locomotor recovery. There are few controlled studies that compare outcomes of BWS training or FES with those of conventional interventions, and access to services using BWS training or FES may be restricted under managed care. PMID- 10792859 TI - A guide for use and interpretation of kinesiologic electromyographic data. AB - Physical therapists are among the most common users of electromyography as a method for understanding function and dysfunction of the neuromuscular system. However, there is no collection of references or a source that provides an overview or synthesis of information that serves to guide either the user or the consumer of electromyography and the data derived. Thus, the purpose of this article is to present a guide, accompanied by an inclusive reference list, for the use and interpretation of kinesiologic electromyographic data. The guide is divided into 4 major sections: collecting, managing, normalizing, and analyzing kinesiologic electromyographic data. In the first of these sections, the issues affecting data collection with both indwelling and surface electrodes are discussed. In the second section, data management through alternative forms of data processing is addressed. In the third section, various reasons and procedures for data normalization are discussed. The last section reviews qualitative descriptors once used as the only means of analyzing data, then focuses on more quantitative procedures that predominate today. The guide is intended as a tool for students, educators, clinicians, and beginning researchers who use and interpret kinesiologic electromyographic data. Modifications will likely be needed as alternative forms of collecting, managing, normalizing, and analyzing electromyographic data are proposed, used in various settings, and reported in the literature. PMID- 10792860 TI - Clinical research agenda for physical therapy. AB - The American Physical Therapy Association (APTA) has developed a clinical research agenda that is designed to support, explain, and enhance physical therapy clinical practice by facilitating research that is useful primarily to clinicians. The Clinical Research Agenda was developed through a series of conferences and extensive editorial and review processes and represents input from a large number of physical therapists. The Clinical Research Agenda represents questions that are believed to be important to clinical practice, the profession, and APTA. The themes of the Clinical Research Agenda were developed in an attempt to span the breadth of patient/client management beyond the particulars of any single question and to signal the full emergence of the physical therapist clinician as a scientific practitioner. Furthermore, the Clinical Research Agenda is intended to serve as a benchmark of the systematic progression of the scientific basis of the profession as a whole. As approved by APTA's Board of Directors, the Clinical Research Agenda will serve as the focal point for the research programs of the Foundation for Physical Therapy, as directed by the Foundation's trustees, and will be shared with other funding agencies and researchers outside of physical therapy as well. PMID- 10792861 TI - Outcomes and aging. PMID- 10792862 TI - Blood donation. PMID- 10792863 TI - Was torque measured? PMID- 10792864 TI - Pelvic asymmetry reliability. PMID- 10792865 TI - Occupational injuries in PTs. PMID- 10792866 TI - Michels as motivator. PMID- 10792867 TI - Competence and complacency. PMID- 10792868 TI - Gynecologic Cancer: Screening, Treatment Options, and Quality-of-Life Considerations. PMID- 10792869 TI - Hormone Replacement Therapy in the Gynecologic and Breast Cancer Patient. AB - Hormone replacement therapy has increased in use as a result of its well established benefits as an effective tool against such consequences as osteoporosis and ischemic heart disease and in the management of menopausal symptoms. However, its possible negative impact on the survival of patients with a previous diagnosis of breast, uterine, or ovarian cancer remains controversial. The risk:benefit analysis of hormone replacement therapy in this setting warrants further investigation. In the meantime, patients should be counseled on the risks, benefits, and contraindications of hormone replacement therapy before embarking on its use. PMID- 10792870 TI - Tamoxifen and Endometrial Cancer. AB - Tamoxifen is commonly used in the management of patients with breast cancer. Clinical trials of tamoxifen involving over 75,000 patients demonstrate an improved recurrence-free and overall survival benefit in both pre- and post menopausal women. Large-scale trials also are evaluating the role of tamoxifen as a chemopreventive agent in women considered to be at high risk for developing breast cancer based on family history. Endometrial cancer is an uncommon complication of tamoxifen therapy. Since the majority of these cancers will be detected at an early stage when they are highly curable, however, the overall benefit of tamoxifen treatment in breast cancer patients outweighs this risk. All women receiving tamoxifen who have a uterus should undergo regular gynecologic examinations. PMID- 10792871 TI - Predicting and Treating the Sexual Difficulties of Gynecologic Cancer Survivors. AB - Women treated for gynecologic cancer represent approximately 45% of cancer survivors. Many report significant sexual difficulties during follow-up, and data suggest these difficulties are direct consequences of their cancer treatments. Health care providers can identify at the time of initial therapy those women who will be at greater risk for problem development. Clinical observations are provided on the development of sexual problems for these women as they attempt to resume sexual intercourse, and the psychologic and behavioral therapies that can be used for treatment are discussed. PMID- 10792873 TI - Cytoreductive Surgery in Ovarian Cancer: Why, When, and How? AB - Retrospective evidence supports the value of optimal cytoreductive surgery in the initial therapy of patients with advanced ovarian cancer. Specialized procedures, including radical pelvic surgery, bowel resection, and diaphragm resections, are frequently necessary to accomplish optimal cytoreduction. Cytoreduction and total gross tumor removal are possible more frequently with new surgical instruments such as the Cavitron ultrasonic surgical aspirator and argon beam laser. Pelvic and periaortic lymph node resection is an important aspect of cytoreductive surgery, and systematic removal of grossly uninvolved lymph nodes may improve survival. Secondary cytoreductive surgery appears to benefit a select group of patients. PMID- 10792872 TI - Screening for Ovarian Cancer. AB - Ovarian cancer is difficult to manage because the disease is most often diagnosed at an advanced stage when survival chances are poor. Early detection of ovarian cancer would increase long-term survival, since effective treatment modalities are available for early-onset disease. Screening with transvaginal ultrasound and serum CA 125 suggests promising results, but studies comparing mortality rates for screened vs unscreened populations are needed, and strategies must be developed for prevention or early diagnosis in order to control this disease process. PMID- 10792875 TI - Indolent Lymphomas: Classic Subtypes and Newer Entities. PMID- 10792874 TI - Frontiers of Ovarian Cancer Therapy. AB - Since the majority of patients with ovarian cancer present with advanced stages of disease, more effective systemic approaches are needed to add to the benefits of surgical staging and debulking. New combinations of taxoids with cisplatin have prolonged survival, and other chemotherapeutic agents are being evaluated. Immunotherapy, including intraperitoneal approaches with monoclonal antibodies, cellular therapies and vaccines, hormone therapy with well-known drugs such as tamoxifen, and gene therapy give promise for the future. PMID- 10792876 TI - Pulmonary Pseudotumors: Infections and Other Causes. PMID- 10792877 TI - Screening For Colon and Rectal Cancer. PMID- 10792879 TI - Speed of onset of regional analgesia in labour: a comparison of the epidural and spinal routes. PMID- 10792880 TI - Low-cost prevention of bacterial contamination during continuous epidural treatment by a minimal dose of an antibiotic in the epidural solution. PMID- 10792881 TI - Epidurals--isn't one enough? PMID- 10792882 TI - Probable seizure after remifentanil in a 4-year-old boy. PMID- 10792883 TI - Consider position of patient to prevent ventilator-associated pneumonia. PMID- 10792884 TI - Safe pre-operative fasting times after milk or clear fluid in children. PMID- 10792886 TI - Cognitive function and surgery. PMID- 10792888 TI - Cerebrospinal fluid filtration in Guillain-Barre syndrome. PMID- 10792889 TI - Dystrophia myotonica and succinylcholine. PMID- 10792891 TI - Refractory hypotension during carcinoid resection surgery. PMID- 10792892 TI - Anaesthesia using a laryngeal mask airway in a patient with osteogenesis imperfecta. PMID- 10792893 TI - Propofol and postoperative pancreatitis. PMID- 10792896 TI - Intra-operative monitoring of cardiac output during massive blood loss. PMID- 10792897 TI - A device for oxygen administration during ophthalmic surgery under local anaesthesia. PMID- 10792898 TI - Should we perform a sniff test? PMID- 10792900 TI - Always have a contingency plan. PMID- 10792901 TI - The ECG pad lottery--it could be you! PMID- 10792903 TI - Extending the life of the laryngeal mask airway. PMID- 10792904 TI - 2-D and 3-D reconstruction for tracheal stenosis. PMID- 10792905 TI - The role of physiotherapy in ventilator-associated pneumonia. PMID- 10792906 TI - Change of position relieving upper airway obstruction. PMID- 10792907 TI - Severe epistaxis--throat pack and airway inspection are essential. PMID- 10792909 TI - Airway management--a current training problem? PMID- 10792910 TI - Remifentanil and propofol for tracheal intubation. PMID- 10792912 TI - Remifentanil and intubating conditions. PMID- 10792914 TI - Laryngeal mask airway and the difficult airway. PMID- 10792915 TI - Difficulty in aspiration of gastric contents--a simple solution. PMID- 10792917 TI - Nausea and vomiting: in somno securitas et commoditas. PMID- 10792918 TI - The defibrillator back paddle: use for treatment of arrhythmias during prone position ventilation. PMID- 10792919 TI - Remifentanil for percutaneous tracheostomies in ITU. PMID- 10792920 TI - Pros and cons of manual lung hyperinflation. PMID- 10792922 TI - Ways to test stochastic dynamic programming models empirically. AB - Stochastic dynamic programming (SDP) models are widely used to predict optimal behavioural and life history strategies. We discuss a diversity of ways to test SDP models empirically, taking as our main illustration a model of the daily singing routine of birds. One approach to verification is to quantify model parameters, but most SDP models are schematic. Because predictions are therefore qualitative, testing several predictions is desirable. How state determines behaviour (the policy) is a central prediction that should be examined directly if both state and behaviour are measurable. Complementary predictions concern how behaviour and state change through time, but information is discarded by considering behaviour rather than state, by looking only at average state rather than its distribution, and by not following individuals. We identify the various circumstances in which an individual's state/behaviour at one time is correlated with its state/behaviour at a later time. When there are several state variables the relationships between them may be informative. Often model parameters represent environmental conditions that can also be viewed as state variables. Experimental manipulation of the environment has several advantages as a test, but a problem is uncertainty over how much the organism's policy will adjust. As an example we allow birds to use different assumptions about how well past weather predicts future weather. We advocate mirroring planned empirical investigations on the computer to investigate which manipulations and predictions will best test a model. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792923 TI - To nest communally or not to nest communally: a review of rodent communal nesting and nursing. AB - Communal nesting, the sharing of parental responsibilities between multiple individuals in a nest, is common in many rodents. Upon first glance, this behaviour seems to be selectively disadvantageous. Communal care not only involves energetic costs, but may also be subject to cheating behaviour. Despite abundant literature, many questions remain regarding advantages gained by females that form nesting groups. I discuss the communal nesting of eusocial, singular and plural breeding rodents. I first clarify the distinction between communal nesting and thermoregulatory huddling. I then review two major groups of hypotheses ('ecological constraints' and 'benefits of philopatry') that are used to explain the occurrence of communal nesting in rodents. Most likely, these hypotheses are not mutually exclusive. Regardless of the main selective pressure(s) acting on communal nesting, the indirect components of inclusive fitness that result from nest sharing most likely influence the formation and maintenance of communal nests. Communal nesting and nursing (the sharing of milk with nonoffspring) are under different selective regimes and thus, must be evaluated separately. I review adaptive and nonadaptive hypotheses for rodent communal nursing. I argue that communal nursing may not be adaptive as mothers may be forced to share milk with nonoffspring in large communal nests (i.e. communal nursing may be a cost associated with communal nesting). In conclusion, I propose directions for future study that may improve our understanding of communal nesting and nursing in the wild. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792924 TI - Male infanticide in captive plains zebra, Equus burchelli. AB - On the assumption that infanticide exists in plains zebra, as reported for horses, Equus caballus, we tested the following hypothesis. Introducing a new zebra male into a herd of breeding females should increase foal mortality in comparison with herds in which the sire of the foals is still present. The younger the foal, the more likely infanticide should be. We collected data from five herds in two zoological gardens in the Czech Republic. We found nine records of infanticide in plains zebra and three cases of abortions that were probably induced by forced copulation. We analysed additional indirect data to investigate the possibility of introduced males causing other abortions. Abortions were three times more likely in herds with introduced males than with only fathers present. Postnatal mortality of the foals was four times greater with introduced males than with fathers. No indication of a sex preference was observed for infanticide by a new male for either abortions or postpartum deaths. When we combined all records involving introduced males, the probability of foal death was greatest when the new male joined the herd just after conception and decreased with increasing time between conception and date of the new male introduction (the chance of a foal surviving was less than 5% just after conception and more than 50% at the time of delivery). Mortality of foals did not depend on whether the new male was introduced before or after the foal was born. Survival increased to more than 60% after the foal reached 1 month of age. Our results suggest that captive plains zebra show the highest occurrence of infanticide reported among ungulates. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792925 TI - Copulation duration and fertilization success in a damselfly: an example of cryptic female choice? AB - Copulation duration is highly variable (0.5-3 h) in the damselfly, Ceriagrion tenellum (Coenagrionidae). Using laboratory experiments, we tested four adaptive hypotheses to explain this variation: the effect of time constraints, in-copula mate guarding, sperm displacement and cryptic female choice. Copulation duration was negatively correlated with time of day, as predicted by the first two hypotheses, and positively correlated with male density, as predicted by the mate guarding hypothesis. Males prolonged copulation in response to the volume of sperm stored by females, suggesting they were able to detect and quantify the amount of sperm stored. This behaviour is not explained by mate guarding or time constraint effects. Males removed all the sperm from the bursa copulatrix in just 10 min. Our results also suggest that, because the duct is too narrow to allow male genitalia to enter, males do not remove spermathecal sperm. Therefore, direct sperm removal could not explain long copulations. Prolonged copulations could also have evolved as a result of cryptic female choice if they increase male fertilization success by female-mediated processes. Our results support this idea: male fertilization success was greater after long copulations. Apparently, male copulatory behaviour elicits female responses that increase male fertilization success. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792926 TI - Estimating the energetic cost of fighting in shore crabs by noninvasive monitoring of heartbeat rate. AB - After establishing shore crabs, Carcinus maenas, individually in separate aquaria, we used a noninvasive infrared phototransducer to monitor their heartbeat rate continously before, during and after fights with intruder crabs. We confirmed that heartbeat rate is a reliable indicator of oxygen consumption and then used it to estimate indirectly the energetic cost of fights differing in duration and intensity, and its dependence on prior residence and relative size of opponent. Prior residence in aquaria significantly increased the probability that crabs would initiate fights against intruders. The majority of fights were resolved by aggressive contacts, display being used extensively only against smaller intruders. Fights between evenly sized opponents and between residents and larger intruders involved almost continuous aggression, whereas fights with smaller intruders involved several shorter bouts of aggression. Fight duration was weakly correlated with the relative size of opponents. Heartbeat rate, measured only in residents, was elevated above resting levels throughout fights, hence energy expenditure during fighting increased linearly with fight duration. Contrary to expectation, heartbeat rate was not significantly influenced by relative size of the opponent or by the intensity of aggression. After fighting, heartbeat rate usually returned to resting levels within 30-60 min, recovery taking longer in fights against larger intruders, when the fight was always lost. We propose that prolonged elevation of heartbeat rate in residents that had lost to larger intruders represented a state of alertness, adaptive against impending risks of resource loss or injury. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792927 TI - Assessment of auditory distance in a territorial songbird: accurate feat or rule of thumb? AB - Territorial passerines presumably benefit from their ability to use auditory cues to judge the distance to singing conspecifics, by increasing the efficiency of their territorial defence. Here, we report data on the approach of male territorial chaffinches, Fringilla coelebs, to a loudspeaker broadcasting conspecific song simulating a rival at various distances by different amounts of song degradation. Songs were degraded digitally in a computer-simulated forest emulating distances of 0, 20, 40, 80 and 120 m. The approach distance of chaffinches towards the loudspeaker increased with increasing amounts of degradation indicating a perceptual representation of differences in distance of a sound source. We discuss the interindividual variation of male responses with respect to constraints resulting from random variation of ranging cues provided by the environmental song degradation, the perception accuracy and the decision rules. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792928 TI - Division of labour and seasonality in the ant Leptothorax albipennis: worker corpulence and its influence on behaviour. AB - We address the organization of workers in social insect societies. We distinguish between changes in behavioural role over the nurse to forager role sequence, which may depend on changes in physiology, and potentially more rapid changes of task within role. We investigated the association between role and nutrient status in the ant Leptothorax albipennis. Worker lipid stores were quantified using a new body size-controlled method, and were related to worker behaviour. Worker lipid stores were evenly distributed amongst colony members at the end of winter, splitting rapidly into two distinct modes (replete nurses and lean foragers) in spring. The proportion of lean foragers increased throughout spring and summer, until most colonies contained only workers of this type. Callow workers then eclosed with intermediate lipid stores. We developed a computer vision system that tracks all nest ants to extract detailed behaviour of individuals of known lipid stores. Lipid storage was negatively correlated with a worker's foraging propensity, and with measures of spatial occupation in the nest and of activity. Different colonies showed a similar quantitative correlation between lipid stores and behavioural role, suggesting that lipid stores were not only correlated with the relative organization of individuals within each nest, but may also have influenced their absolute role. We reviewed the literature and found evidence that nutrient status influences role predisposition in social insect workers. We conclude that the distribution of worker roles may be linked to the balance between foraging income and energetic consumption within the colony directly via worker nutrient status. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792929 TI - Food allocation in crimson rosella broods: parents differ in their responses to chick hunger. AB - Food allocation in many asynchronously hatching bird species favours large, competitively superior chicks. In contrast, food is usually distributed equally within broods of crimson rosellas, Platycercus elegans, implying that parents do not simply feed the most competitive chick. We used two temporary removal experiments to manipulate hunger of: (1) individual first- or last-hatched chicks, or (2) the whole brood. When only a single chick was hungry, parents compensated fully and chicks gained the same mass over the day as during controls. Mothers and fathers, however, responded in different ways to chick hunger. Mothers did not strongly alter their food allocation when a single chick was hungry, and controlled the distribution of food by refusing to feed first hatched chicks when they were hungry and by moving more during feeds. In contrast, fathers allocated more food to hungry last-hatched chicks. When the whole brood was hungry, parents were unable to compensate chicks and all chicks lost mass over the day. In these conditions, mothers preferentially fed first hatched chicks, while fathers fed all chicks equally. Our results show that both mothers and fathers were able to discriminate and selectively feed chicks, but that parents responded differently to changes in chick hunger within the brood. Fathers responded more strongly to variation in chick hunger within the brood, suggesting they reallocate food based on short-term changes in hunger. Mothers distributed food preferentially to last-hatched chicks except when the whole brood was hungry, when they switched to favouring first-hatched chicks. This pattern is consistent with a strategy of adaptive brood reduction when food is scarce. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792930 TI - Male-male competition and large size mating advantage in European earwigs, Forficula auricularia. AB - European earwigs are sexually dimorphic in forceps shape and length. Male forceps are thought to be weapons in male contests for access to females, but recent findings suggest that females choose males on the basis of their forceps length. I investigated sexual selection on forceps length and body size and the occurrence of male-male competition. When I controlled for forceps length experimentally and statistically, relatively heavy males had greater copulation success than relatively light males. When I controlled for body size, males with relatively longer forceps had no tendency for greater copulation success than males with shorter forceps. Relatively heavy males more often took over copulations from smaller males than vice versa. Male contests were important for the outcome of mate competition, as males commonly interrupted and took over copulations. My results therefore suggest that intrasexual selection is significant in competition for copulations in male earwigs, and acts on body size. This contrasts with previous findings, which have shown intersexual selection on forceps length to be important. However, both modes of sexual selection may be acting through a two-stage process, where male-male competition first determines which males have access to females, and then through female choice among available males. Morphological measurements supported the conclusion that forceps length and body size are male secondary sexual characters, as these characters had large variance and skewed distributions in males, but were normally distributed in females. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792931 TI - Female baboons do not raise the stakes but they give as good as they get. AB - We used data from four chacma baboon, Papio cynocephalus ursinus, troops, living in two populations, to test the raise the stakes (RTS) strategy of reciprocity. Female baboons did not raise the stakes either within or across grooming bouts. Instead they time-matched grooming contributions and divided grooming into short episodes. In addition, analysis of the grooming behaviour of frequently versus infrequently grooming dyads did not reveal differences in grooming patterns predicted by the RTS strategy. We suggest time constraints preclude the escalation of grooming bout length as required by RTS; the data were more consistent with a strategy of give as good as you get. However, this strategy could not explain all the patterns observed, and we conclude that biological market theory represents a more appropriate framework for investigating female grooming dynamics than dyadic games based on the iterated prisoner's dilemma. We suggest that competitive altruism among individuals acts as a market force influencing an individual's value as a grooming partner. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792932 TI - Chimpanzees know what conspecifics do and do not see. AB - We report a series of experiments on social problem solving in chimpanzees, Pan troglodytes. In each experiment a subordinate and a dominant individual were put into competition over two pieces of food. In all experiments dominants obtained virtually all of the foods to which they had good visual and physical access. However, subordinates were successful quite often in three situations in which they had better visual access to the food than the dominant, for example, when the food was positioned so that only the subordinate (and not the dominant) could see it. In some cases, the subordinate might have been monitoring the behaviour of the dominant directly and simply avoided the food that the dominant was moving towards (which just happened to be the one it could see). In other cases, however, we ruled out this possibility by giving subordinates a small headstart and forcing them to make their choice (to go to the food that both competitors could see, or the food that only they could see) before the dominant was released into the area. Together with other recent studies, the present investigation suggests that chimpanzees know what conspecifics can and cannot see, and, furthermore, that they use this knowledge to devise effective social-cognitive strategies in naturally occurring food competition situations. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792933 TI - Strategic diel regulation of body mass in European robins. AB - Stochastic dynamic programming (SDP) is a computational technique that has been used to model daily routines of foraging in small birds. A diurnal bird must build up its fat reserves towards dusk in order to avoid starvation during the night, when it cannot feed. However, as well as the benefits of avoiding starvation, storing fat imposes costs such as an increased predation risk and higher flight and metabolic costs. There is therefore an optimal level of fat reserves for a bird to reach at dusk in order to survive overnight without being left with excessive fat reserves at dawn. I tested a prediction common to all SDP models of daily foraging routines, that a bird will attempt to reach this level at dusk, regardless of its fat reserves the previous dawn. I provided supplementary food to manipulate the fat reserves at dawn of free-living European robins, Erithacus rubecula. Diurnal changes in body mass (a reliable estimate of fat reserves) were then monitored remotely. Robins provided with an ad libitum food supply reached almost exactly the same body mass at dusk, regardless of their body mass at dawn, supporting the prediction that birds attempt to reach a target level of reserves at dusk. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792934 TI - Temporal shift in the presence of a chemical cue contributes to a diel shift in sociality. AB - Spatial and temporal variation in animal aggregations may be due to variation in the presence of cues for aggregation (or disaggregation) or to variation in the receptivity of the animal to a particular cue or suite of cues. Spiny lobsters, Panulirus argus, forage solitarily but are often found aggregated in their diurnal shelters. An important proximate cause of aggregation among spiny lobsters is a scent they produce that influences shelter choice by conspecifics. We examined how variability in the presence of, or response to, such a chemical cue may contribute to diel shifts in sociality among spiny lobsters. We conducted a series of Y-maze shelter choice experiments using lobsters that were either maintained under altered dark:light schedules in the experimental arena or under natural lighting in the head tanks. Lobsters that were maintained on a light schedule 8 h later than normal chose shelters at their dawn (corresponding to the middle of the night for lobsters in the head tanks); however, their choices of shelter were not influenced by scents of conspecifics. Lobsters that were maintained on a schedule 8 h earlier than normal chose shelters in the middle of their night (corresponding to dawn for the lobsters in the head tanks). Their choices of shelter were significantly influenced by conspecific scents. These results suggest that the chemical cues for aggregation, released by spiny lobsters, are present discontinuously, that spiny lobsters are influenced by conspecific odours continuously, and that aggregation is controlled by temporal variation in the presence of a chemical cue. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792935 TI - Why some male Mongolian gerbils may help at the nest: testosterone, asexuality and alloparenting. AB - In previous studies we have shown that those male Mongolian gerbils, Meriones unguiculatus, that as fetuses resided in intrauterine positions (IUPs) located between two female fetuses (2F males) have lower circulating levels of testosterone, less well-developed genital musculature, and lower reproductive success than males gestated in IUPs between two male fetuses (2M males). We have also found that such 2F males spend more time caring for conspecific young than do 2M males, and that presence of a 2F male, but not of a 2M male, with a lactating female and her litter decreases the cost to a dam of suckling one litter while gestating a second litter conceived in postpartum oestrus. Here we show that some 2F males, those with circulating levels of testosterone similar to those seen in females of their species, show no interest in females in oestrus and fail to impregnate females with which they are paired. Such 'asexual' 2F males spend 30-50% more time caring for nestlings than do sexually active 2F males. We suggest that such asexual, highly parental 2F males are incapable of direct reproduction and are obligate helpers at the nest that can contribute to their own fitness only by assisting to rear collateral kin. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792936 TI - The neuropeptide arginine vasotocin alters male call characteristics involved in social interactions in the grey treefrog, Hyla versicolor. AB - We investigated the effects of different doses (0, 2.5, 25 and 250 ug) of the neuropeptide arginine vasotocin (AVT) on the calling characteristics of the grey treefrog in a chorus in its natural habitat. AVT changed some call characteristics known to influence social behaviour in grey treefrogs. It increased call duration and number of pulses in a call, but not dominant frequency, call rate or pulse effort. Saline injections and handling did not produce significant changes in any of the call characteristics. In addition, individual animals injected with AVT only rarely produced call characteristics that were outside of the range found in the preinjection measurements, suggesting that AVT does not cause abnormal calling behaviour. Other researchers have demonstrated that longer calls with more pulses are produced by males when chorus densities increase, and females display a strong preference for longer calls with more pulses. This suggests that the changes induced by AVT injections may have functional consequences in social interactions. We previously demonstrated that AVT-injected males (25 ug AVT) displaced resident males from calling sites through changes in calling behaviour under natural field conditions. Our results indicate that changes in call duration and pulse number could contribute to the unmanipulated resident male's behaviour towards the AVT-injected intruder, perhaps because the calls are more attractive to females or because the calls are perceived as more aggressive. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792937 TI - Aggressiveness in king penguins in relation to reproductive status and territory location. AB - King penguins, Aptenodytes patagonicus, vigorously defend small territories in very dense colonies. The egg-laying season lasts approximately 4 months, but only pairs that reproduce during the first half of the period succeed in fledging a chick. I examined various factors affecting aggressiveness of king penguins during the breeding season and focused on the differences between central and peripheral territories. Pairs on peripheral territories experienced twice as many encounters with avian predators as did central birds. The vast majority of peripheral birds were late breeders, indicating that reproductive success was very low among penguins defending a territory on the edge of the colony. Time invested in territory defence and rate of agonistic encounters between breeding neighbours increased from the incubation to the brooding period. Parents gave most threat displays to territorial neighbours when the chick was very young and just before creche formation. Distance to colony edge was not related to aggressiveness in incubating birds, however, the rate of pecking and flipper blows increased from the edge to the centre during brooding. In addition, aggressiveness of breeding penguins towards travelling birds trespassing into their territory increased with distance to edge. Early breeders were not more aggressive than late breeders but the proportion of time spent in territory defence increased with the number of days a bird had spent incubating. As expected, I did not detect any sex difference in aggressive behaviour. Birds occupying territories on the beach were generally more aggressive during the incubation period than those located on the valley sides. Reproductive status (incubating versus brooding) and territory location were the main factors explaining the various levels of aggressiveness observed in breeding king penguins. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792938 TI - Nest desertion and cowbird parasitism: evidence for evolved responses and evolutionary lag. AB - Nest desertion with subsequent renesting is a frequently cited response to parasitism by the brown-headed cowbird, Molothrus ater, yet the role of desertion as an antiparasite defence is widely debated. To determine whether desertion represents an evolutionary response to brown-headed cowbird parasitism, we searched the primary literature, yielding data on the desertion frequencies of 60 host populations from 35 species. Species were categorized according to three habitat types (forest, intermediate and nonforest). Because cowbirds prefer open habitat and rarely penetrate deeply into forests, nonforest species have long been exposed to widespread cowbird parasitism, whereas forest species have not. However, due to increased forest fragmentation, forest species are being increasingly exposed to extensive parasitism. The frequency of desertion of parasitized nests was significantly higher in nonforest than forest species, suggesting that the latter experience evolutionary lag. We also considered whether desertion is affected by predation frequency, degree of current or recent sympatry with cowbirds, parasitism frequency, length of host laying season, phylogenetic relationships, and potential cost of cowbird parasitism. None of these variables created biases that could account for the observed difference in desertion frequencies of nonforest and forest species. However, species that incur large costs when parasitized had higher desertion rates among nonforest species but not among forest species. These results indicate that increased nest desertion is an evolved response to cowbird parasitism, as one would otherwise expect no relationship between desertion frequency and thezx costs and length of exposure to cowbird parasitism. Although nearly all hosts have eggs easily distinguished from cowbird eggs, few or none desert in response to cowbird eggs. Instead, desertion may be a response to adult cowbirds. The scarcity of species that desert in response to cowbird eggs suggests that egg recognition is more difficult to evolve than heightened desertion tendencies and that egg recognition quickly leads to ejection behaviour once it does develop. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792939 TI - The role of queens in colonies of the swarm-founding wasp Parachartergus colobopterus. AB - Social insect queens reproduce while workers generally do not. Queens may also have other behavioural roles in the colony. In small, independent-founding colonies of social wasps, the dominant queen physically enforces her interests over those of the workers and serves as a pacemaker of the colony, stimulating workers to forage and engage in other tasks. By contrast, in large-colony, swarm founding wasps, the collective interests of the workers are fulfilled in sex allocation and production of males, whether or not they coincide with the interests of the queens. The behavioural role of the queens in such species has not been extensively studied. We investigated the role of the queens both in regulating worker activity and in reducing the numbers of reproductively active queens in the swarm-founding epiponine wasp Parachartergus colobopterus. We found no evidence that queens regulate worker activity, as they were rarely involved in any interactions. Worker activity may be self-organized, without centralized active control by anyone. Furthermore, we found no evidence that the reduction in queen number characteristic of this tribe of wasps occurs in response to aggression among queens. The reduction in queen number may be a result of worker treatment of queens, although worker discrimination against some queens was not obvious in our data. i Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792940 TI - Lack of pubertal influences on female dispersal in muriqui monkeys, Brachyteles arachnoides. AB - The hormonal mediation of dispersal in female mammals is poorly understood, in part because of the difficulties of detecting the onset of ovarian cycling and puberty in dispersing individuals. We used noninvasive methods of faecal steroid assays to determine the timing of dispersal relative to puberty and ovarian cycling in wild female muriqui monkeys, a species in which males are philopatric and nearly all females transfer from their natal groups. Natal females had a mean+/-SE age of 73.4+/-7.2 months (N=18) at the time of their transfers. Intergroup transfers occurred when one or more sexually active adult females were present, but did not show any seasonal patterns. Faecal progesterone and oestradiol profiles from nine natal females prior to transfer and four non-natal females that transferred into our study group demonstrate unequivocally that dispersal occurs prior to puberty in this species. All females showed baseline oestradiol levels and low progesterone levels compared with cycling adult females. Immigrants were first observed to copulate at 11.2+/-2.2 months of age (N=4), prior to the onset of normal ovarian cycles, and gave birth to their first offspring at 33.8+/-7.3 months (N=4) after transferring. Mean cortisol levels did not differ between natal emigrants or recent immigrants, and were within the range of those of adult males during the nonbreeding season in 10 of the 11 prepubertal females sampled. These results indicate that female dispersal is not triggered by activational hormones associated with puberty or escape from reproductive suppression in this species. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792941 TI - Is there an intraspecific role for density-dependent colour change in the desert locust? AB - Attempts to uncover the adaptive significance of density-dependent colour polyphenism in the desert locust, Schistocerca gregaria (Orthoptera: Acrididae), have been unsuccessful. Desert locust juveniles can change colour as part of a phenotypically plastic response to changes in local population density known as phase polyphenism. They are typically cryptic in colour at low rearing density (solitarious phase), but become conspicuous at high density (gregarious phase). Recent evidence indicates that this colour change functions interspecifically as an aposematic signal. Other recent evidence, however, suggests that previous attempts to demonstrate an intraspecific function of gregarious coloration in mediating group interactions among locusts may have been confounded by the effects of multiple sensory cues. We reinvestigated the intraspecific function of density-dependent colour polyphenism and specifically controlled for potentially confounding olfactory and tactile cues. We found no effect of gregarious phase (yellow and black) coloration as either a gregarizing stimulus to behaviourally solitarious locusts or as a visual aggregation stimulus behaviourally to gregarious locusts. We did, however, find that nonmoving solitarious phase (green) coloration significantly increased the activity levels of behaviourally gregarious locusts. We cannot explain this result and its biological relevance remains unknown. In the absence of support for the intraspecific visual cue hypothesis, we favour an aposematic perspective on the function of density dependent colour polyphenism in the desert locust. The aposematic perspective parsimoniously accounts for density-dependent changes in both colour and behaviour. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792942 TI - Juvenile western toads, Bufo boreas, avoid chemical cues of snakes fed juvenile, but not larval, conspecifics. AB - Previous investigations have demonstrated the importance of predator diet in chemically mediated antipredator behaviour. However, there are few data on responses to life-stage-specific predator diets, which could be important for animals like amphibians that undergo metamorphosis and must respond to different suites of predators at different life-history stages. In laboratory choice tests, we investigated the chemically mediated avoidance response of juvenile western toads, Bufo boreas, to four different chemical stimuli: (1) live conspecific juveniles; (2) live earthworms; (3) snakes fed juvenile conspecifics; and (4) snakes fed larval conspecifics (tadpoles). Juvenile toads avoided chemical cues from snakes that had eaten juvenile conspecifics, but did not respond to the other three stimuli, including chemical cues from snakes fed larval conspecifics. In addition, the response to cues from snakes fed juveniles differed significantly from that of snakes fed larvae. To our knowledge, this is the first study to demonstrate the importance of diet in predator avoidance of juvenile anurans and the ability of juvenile toads to distinguish between chemical cues from predators that have consumed larval versus juvenile conspecifics. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792943 TI - Egg rejection in a passerine bird: size does matter. AB - Avian brood parasites reduce the reproductive success of their hosts, selecting for the evolution of egg discrimination by the host, and potentially creating a coevolutionary arms race between host and parasite. Host egg discrimination ability is crucial in determining whether the arms race results in extinction (of the parasite on a particular host) or stable coevolutionary equilibrium of the host-parasite pair. I examined egg discrimination behaviour in the yellow-browed leaf warbler, Phylloscopus humei, a presumed former host of parasitic cuckoos, to show how discrimination ability has become very strong. Field experiments using model eggs demonstrate that rejection decisions are based on the relative size of eggs in the clutch. Individuals do not learn the particular size of their own eggs, but will accept both large and small eggs as long as all eggs in the clutch are of similar size. Host rejection decisions are continuously modified based on assessment of variation in egg sizes currently in the clutch, making it a difficult strategy for a cuckoo to defeat. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792945 TI - Mother Nature: A History of Mothers, Infants, and Natural Selection. PMID- 10792944 TI - Male affiliation, cooperation and kinship in wild chimpanzees. AB - Long-term field research has revealed that male chimpanzees, Pan troglodytes, affiliate and cooperate in several contexts. Assuming close genetic relationship among males, affiliative and cooperative behaviour have been hypothesized to evolve through the indirect effects of kin selection. We tested the hypothesis that matrilineal genetic relatedness affects patterns of male social affiliation and cooperation in an unusually large community of chimpanzees at the Ngogo study site, Kibale National Park, Uganda. Field observations indicated that six behavioural measures of affiliation and cooperation among 23 adult males were significantly correlated with each other. Sequences of the first hypervariable portion of the mtDNA genome revealed that three pairs of males and one quintet shared mtDNA haplotypes. Matrix permutation tests using behavioural and genetic data showed that males that affiliated and cooperated with each other were not closely related through the maternal line. These findings add to a growing body of empirical evidence that suggest kinship plays an ancillary role in structuring patterns of wild chimpanzee behaviour within social groups. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 10792946 TI - Information Processing in Social Insects. PMID- 10792947 TI - Elastase mimics pancreatitis-induced hepatic injury via inflammatory mediators. AB - BACKGROUND: Recent evidence suggests that pancreatitis-associated hepatic injury is regulated by inflammatory mediator production. Our laboratory demonstrated in vitro that pancreatic elastase is a pancreatic enzyme that can induce inflammatory cell cytokine production. Therefore we now explore the in vivo effects of elastase on the liver. MATERIALS AND METHODS: Elastase (1.5 U) +/- CNI 1493, which attenuates mediator production through p38 MAP kinase inhibition, was administered intraperitoneally to mice while control animals received saline. Acute pancreatitis (AP) was induced with a choline-deficient, ethionine supplemented (CDE) diet. Serum hepatic enzymes and hepatic neutrophil infiltration by myeloperoxidase (MPO) activity were measured as indicators of hepatic insult. Serum tumor necrosis factor (TNF) protein (ELISA), hepatic TNF mRNA (reverse transcription polymerase chain reaction), and hepatic activation of the transcription factor nuclear factor kappa B (electrophoretic mobility shift assay) were also determined. RESULTS: A significant increase in hepatic enzymes and MPO activity was induced by AP and mirrored by intraperitoneal elastase. Both types of liver injury resulted in near identical elevations in serum TNF protein and hepatic TNF mRNA. Elastase-treated animals with mediator production inhibited (CNI-1493) had attenuated hepatic enzymes, MPO activity, TNF protein, and TNF mRNA. Activation of nuclear factor kappa B occurred 30 min after elastase administration. CONCLUSION: Exposure of the liver to pancreatic elastase results in hepatic inflammation and injury which appears identical to that seen during severe AP. Prevention of inflammatory mediator production by intrahepatic leukocytes attenuates injury and supports recent adult respiratory distress syndrome and in vitro data suggesting that elastase is the principal factor that propagates pancreatic inflammation into a systemic illness through direct activation of systemic inflammatory cells. PMID- 10792949 TI - The effects of interleukin-10 in hemorrhagic shock. AB - BACKGROUND: Interleukin-10 (IL-10) counteracts the effects of the proinflammatory cytokines interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF). Experimental data suggest that inhibition of these proinflammatory cytokines improves outcome in sepsis, endotoxemia, necrotizing pancreatitis, and other severe inflammatory states. We hypothesized that the administration of IL 10 would attenuate the release of proinflammatory cytokines after severe hemorrhagic shock. METHODS: To test our hypothesis, male Sprague-Dawley rats (N = 20) were divided into control and experimental groups. We induced hemorrhagic shock by removing a sufficient quantity of blood to maintain a mean arterial pressure of 50 mm Hg or less for 120 min. The animals were then resuscitated with shed blood and an equal volume of 0.9% saline. The experimental group received 10,000 units of IL-10 at the initiation of shock. Serum IL-1, IL-6, TNF, and lactate were measured at baseline, after 120 min of shock, and 60 min after resuscitation. The rats were followed for 72 h to calculate survival. RESULTS: Similar levels of hypoperfusion were obtained in both groups as demonstrated by lactate levels and amount of shed blood. The survival rate (70%) was the same in both groups. Serum levels of IL-1 and IL-6 were not significantly different between the two groups, although there was a trend toward IL-6 suppression. TNF, however, was significantly lower in the IL-10-treated group at the end of shock (Wilcoxon test, P < 0. 025). CONCLUSION: Administration of IL-10 suppresses the TNF surge observed after severe hemorrhagic shock. PMID- 10792948 TI - Dose response to a single intramuscular injection of recombinant adeno-associated virus-erythropoietin in monkeys. AB - BACKGROUND: Anemia is a significant problem in many disease states. Erythropoietin (Epo) has been used in the treatment of anemia associated with numerous chronic diseases. This study investigates the dose-response profiles of a single intramuscular (im) injection of a recombinant adeno-associated virus vector (rAAV) containing the Epo gene with the goal of achieving a sustained elevation of hematocrit (Hct). METHODS: Cynomolgus (cm) monkeys were given single injections of different doses of rAAV-cm-Epo. The biological effect of Epo gene expression was monitored by determining the Hct levels and circulating hormone levels by ELISA. Antibody to the rAAV capsid protein was also measured over the 41-week period of the experiment. RESULTS: Epo expression was noted only when 2 x 10(11) or more particles were injected. Epo was noted to be increased as soon as 1 week postinjection and was maximum in 6 to 8 weeks. This level of expression remained constant for nearly 20 weeks. Animals given the highest dose of rAAV developed a higher Hct over the first 8 weeks postinjection than those given an intermediate dose. However, the maximum levels of hemoglobin were the same. There was a weak correlation between amount of rAAV injected and capsid antibody response. CONCLUSIONS: AAV vectors are able to transduce skeletal muscle and are capable of achieving sustained expression and systemic delivery of a therapeutic protein following a single im administration. Dose responses to rAAV-Epo are achievable, although a threshold inoculum of virus is necessary to produce an effect and the therapeutic window is narrow. PMID- 10792950 TI - Neuroblastoma and hepatocyte coculture conditioned media alter apoptosis. AB - BACKGROUND: Neuroblastoma is a childhood tumor that often displays unusual biological behavior. The tumor may present with widespread metastases that are unresponsive to aggressive treatment. At other times, both the metastases and the primary tumor may spontaneously regress without treatment. Apoptosis, or programmed cell death, is thought to play a role in the dichotomous behavior of neuroblastoma. We hypothesize that neuroblastoma cells will interact with host tissues to release mediators that affect apoptosis. MATERIALS AND METHODS: Human neuroblastoma cells and human Chang hepatocytes are grown in a noncontact, coculture system. After incubation for 4 days, the medium from the coculture system is collected. Neuroblastoma cells and Chang hepatocytes are then plated separately with the conditioned medium and their own standard growth medium as controls. After 4 days, these cells are harvested and cytospins made for immunostaining. Tumor necrosis factor alpha (TNF-alpha), Fas ligand, and Bcl-2, are measured with immunohistochemistry. Apoptosis is detected with the TUNEL method. Immunostaining data are interpreted with computer image analysis and reported as stain index. TUNEL data are reported as percentage apoptotic cells. All data are reported as means +/- SEM. Statistical analysis is performed and P < 0.05 considered significant. RESULTS: Chang hepatocytes grown in the coculture conditioned media have an increase in TNF-alpha and Fas ligand. The neuroblastoma cells have a significant decrease in Fas ligand. There is a significant increase in the number of apoptotic hepatocytes when they are cultured in the conditioned media. In contrast, the neuroblastoma cells grown in the coculture conditioned media show no increase in apoptosis. Finally, Bcl-2 is significantly increased in the neuroblastoma cells cultured in the conditioned media. CONCLUSIONS: Neuroblastoma cells grown in coculture conditioned media show increased expression of Bcl-2 and decreased Fas ligand levels. These changes should diminish apoptosis activity in the tumor cells. In contrast, the conditioned media induce elevated levels of proapoptotic mediators in the Chang hepatocytes. A tumor's ability to successfully metastasize may be dependent on mediators generated in the tumor-host interaction, and may not be just an independent characteristic of the tumor itself. PMID- 10792951 TI - Mechanism of delayed rejection in transgenic pig-to-primate cardiac xenotransplantation. AB - BACKGROUND: Pig-to-primate cardiac xenografts undergo hyperacute rejection (HAR), in which primate IgM bind to porcine endothelial alpha-Gal molecules and activate membrane attack complex (MAC) deposition. Prolonged graft survival can be achieved by using transgenic pig donors, which express human complement regulatory proteins (hCRP) to inhibit MAC. However, these xenografts invariably fail from delayed xenograft rejection (DXR). We sought to investigate the poorly understood DXR process. MATERIALS AND METHODS: Wild-type (n = 3) and transgenic (n = 3) porcine hearts were heterotopically transplanted into baboons. Biopsies were analyzed by histology and by immunohistochemistry for porcine endothelial markers (vWF, alpha-Gal, and beta-Gal) and primate IgM and MAC deposition. RESULTS: Wild-type xenografts survived 60-80 min but succumbed to rapid IgM/MAC deposition and microvascular thrombosis. Transgenic xenografts avoided HAR but showed increasing IgM/MAC deposition before rejection on days 5, 7, and 11. Serum from baboons after transgenic xenograft rejection showed increased activity against porcine endothelial cells, and in vitro incubation of untransplanted porcine cardiac sections with sensitized baboon serum showed elevated microvascular IgM binding. Increased IgM deposition appeared specific to alpha Gal, since it competes specifically with alpha-Gal-specific GS-4 lectin, but not with beta-Gal-specific RCA-1 lectin. Competition with GS-4 was not seen if naive baboon serum was used. CONCLUSION: DXR may be mediated by increasing baboon IgM binding on porcine microvascular endothelial alpha-Gal molecules. PMID- 10792952 TI - In vitro validation of duct differentiation in developing embryonic mouse pancreas. AB - BACKGROUND: Early embryonic pancreatic epithelia have the capacity for either endocrine or exocrine lineage commitment. Recent studies demonstrated the pluripotential nature of these undifferentiated cells. Isolated pancreatic epithelia grown under the renal capsule formed primarily islets. However, when these same epithelia were grown in a basement-membrane-rich gel (Matrigel) they formed mostly ducts. Currently, there is no model for in vitro pancreatic duct formation and therefore, the mechanism of duct morphogenesis has never been described. The purpose of this study was to provide such a model by characterizing the expression of two duct markers, carbonic anhydrase II (CAII) and the cystic fibrosis transmembrane conductance regulator (CFTR), in isolated undifferentiated pancreatic epithelia grown in vitro. MATERIALS AND METHODS: We microdissected embryonic pancreases at Embryonic Days (E)9.5-11.5 and performed RT-PCR for CAII and CFTR on E9.5 whole pancreases, E10. 5 and E11.5 epithelia, as well as E11.5 epithelia grown for 7 days in Matrigel. Next we performed in situ hybridization for CAII and CFTR and immunohistochemistry for CAII on E11.5 epithelia grown for 7 days in Matrigel. RESULTS: Early, undifferentiated embryonic pancreatic epithelium does not express CAII and CFTR by RT-PCR. When E11.5 epithelia were grown for 7 days in Matrigel, however, gene expression for both markers is upregulated as ducts form. Furthermore, CAII was seen by IHC and both CAII and CFTR were seen by in situ hybridization in the ducts after 7 days in Matrigel. CONCLUSIONS: These data validate our in vitro system as a model for studying the mechanism of normal pancreatic duct differentiation and may potentially help us to understand the faulty mechanism involved in pancreatic ductal carcinogenesis. PMID- 10792953 TI - Pinacidil pretreatment extends ischemia tolerance of neonatal rabbit hearts. AB - OBJECTIVES: Activating ATP-sensitive potassium (K(ATP)) channels improves ischemia tolerance of adult rabbit hearts. We hypothesize that (a) endogenous activation of the K(ATP) channel accounts for better ischemia tolerance of neonatal hearts and (b) exogenous K(ATP) channel activation with pinacidil further improves the neonatal heart's tolerance to cardioplegic ischemia. METHODS: Study 1: Seven (control) neonatal rabbits received intraperitoneal saline, whereas five others (Glib) received 0.3 mg/kg glibenclamide 10 min before sacrifice. They were perfused on Langendorff with Krebs-Henseleit buffer (KHB). Baseline left ventricle (LV) performance and coronary flow (CF) were measured. After 20 min of 37 degrees C ischemia and 10 min of reperfusion, recovery was measured. Study 2: Ten (control) neonatal hearts underwent 90 min of normothermic ischemia with St. Thomas' cardioplegia (STCP) solution administered every 30 min. Ten others were pretreated with a 10-min infusion of 1 microM pinacidil in KHB and received 1 microM pinacidil-enriched STCP. Recovery of LV performance and CF were measured after 60 min of reperfusion. RESULTS: Study 1: Glib significantly reduced preischemia LV performance by 28%* compared to control hearts. Recovery of Glib-treated hearts was significantly less (67%*) than controls (81%*). Study 2: Pinacidil-treated hearts had significantly better recovery of LV performance (39%*) and CF (78%*) compared to 23 and 52%, respectively, in untreated controls (*P < 0.05 vs control hearts). CONCLUSIONS: Endogenous K(ATP) channel activation in neonatal hearts contributes to their better tolerance to ischemia. Exogenous K(ATP) channel activation by pinacidil pretreatment and cardioplegic enrichment significantly improved the neonatal rabbit heart's tolerance to cardioplegic ischemia. This may be an important addition to myocardial protection during pediatric cardiac surgery. PMID- 10792955 TI - Improved biochemical preservation of lung slices during cold storage. AB - BACKGROUND: Development of lung preservation solutions typically requires whole organ models which are animal and labor intensive. These models rely on physiologic rather than biochemical endpoints, making accurate comparison of the relative efficacy of individual solution components difficult. We hypothesized that lung slices could be used to assess preservation of biochemical function during cold storage. MATERIALS AND METHODS: Whole rat lungs were precision cut into slices with a thickness of 500 microm and preserved at 4 degrees C in the following solutions: University of Wisconsin (UW), Euro-Collins (EC), low potassium-dextran (LPD), Kyoto (K), normal saline (NS), or a novel lung preservation solution (NPS) developed using this model. Lung biochemical function was assessed by ATP content (etamol ATP/mg wet wt) and capacity for protein synthesis (cpm/mg protein) immediately following slicing (0 h) and at 6, 12, 18, and 24 h of cold storage. Six slices were assayed at each time point for each solution. The data were analyzed using analysis of variance and are presented as means +/- SD. RESULTS: ATP content was significantly higher in the lung slices stored in NPS compared with all other solutions at each time point (P < 0.0001). Protein synthesis was significantly higher in the lung slices stored in NPS compared with all other solutions at 6, 12, and 18 h of preservation (P < 0.05). CONCLUSIONS: This lung slice model allows the rapid and efficient screening of lung preservation solutions and their components using quantifiable biochemical endpoints. Using this model, we have developed a novel solution that improves the biochemical preservation of lung slices during cold storage. PMID- 10792954 TI - Increased pulmonary vascular contraction to serotonin after cardiopulmonary bypass: role of cyclooxygenase. AB - BACKGROUND: Pulmonary vascular resistance is frequently elevated after cardiopulmonary bypass (CPB). We examined if altered pulmonary microvascular reactivity to serotonin (5-HT) is due to altered expression of isoforms of nitric oxide synthase (NOS) or cyclooxygenase (COX). MATERIALS AND METHODS: Pigs (n = 8) were heparinized and placed on total CPB for 90 min and then perfused off CPB for 90 min. Noninstrumented pigs (n = 6) served as controls for vascular studies. Relaxation responses (% of precontraction) of microvessels (60-150 microm in diameter) were examined in vitro in a pressurized (20 mm Hg) no-flow state with video microscopic imaging. Expression of eNOS, iNOS, and inducible (COX-2) and constitutive (COX-1) cyclooxygenase was examined with Western blotting and reverse transcription polymerase chain reaction. RESULTS: Pulmonary vascular resistance (PVR) increased from 316 +/- 39 mm Hg x s/cm(5) at baseline to 495 +/- 53 at 60 min and 565 +/- 62 at 90 min after termination of CPB. 5-HT elicited a relaxation response (46.8 +/- 11. 8%) in precontracted control microvessels. This response was not affected by the NOS inhibitor N(G)-nitro-l-arginine. After CPB, pulmonary microvessels contracted significantly to 5-HT (-29 +/- 27%, P < 0.05 vs control). This response was partially inhibited (7 +/- 20%, P = 0.06) in the presence of the COX-2 inhibitor NS398, but was unaffected by the thromboxane synthase inhibitor U63557A (-20 +/- 19%). Expression of iNOS or COX-1 was not changed after CPB. Protein and mRNA expressions of COX-2 both increased significantly after CPB, while that of eNOS decreased by approximately 50%. CONCLUSIONS: PVR increased after CPB. This was associated with a hypercontractile response of isolated pulmonary microvessels to 5-HT that was in part mediated by the release of prostaglandins (but not thromboxane) and associated with increased expression of COX-2 and with decreased expression of eNOS. PMID- 10792956 TI - Phorbol esters rapidly attenuate glutamine uptake and growth in human colon carcinoma cells. AB - BACKGROUND: The amino acid glutamine, while essential for gut epithelial growth, has also been shown to stimulate colon carcinoma proliferation and diminish differentiation. Human colon carcinomas are known to extract and metabolize glutamine at rates severalfold greater than those of normal tissues, but the regulation of this response is unclear. Previously we reported that phorbol esters regulate hepatoma System ASC/B(0)-mediated glutamine uptake and cell growth. As human colon carcinoma cells use this same transporter for glutamine uptake, the present studies were undertaken to determine whether similar regulation functions in colon carcinoma. MATERIALS AND METHODS: Human colon carcinoma cell lines (WiDr and HT29) were treated with the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) and initial-rate transport of glutamine and other nutrients was measured at specific times thereafter. Growth rates were monitored during culture +/- PMA or an excess of System ASC/B(0) substrates relative to glutamine. RESULTS: PMA treatment induced a rapid inhibition of glutamine uptake rates in WiDr and HT29 cells by 30 and 57%, respectively, after 1 h. Cycloheximide failed to block this response, indicating that the mechanism by which PMA exerts its effects is posttranslational. The inhibition of glutamine uptake by PMA was abrogated by the PKC inhibitor staurosporine, suggesting that this rapid System ASC/B(0) regulation may be mediated by a PKC-dependent pathway. PMA also significantly decreased transport via System y(+) (arginine) and System A (small zwitterionic amino acids). Chronic phorbol ester treatment inhibited WiDr cell growth, as did attenuation of System B(0)-mediated glutamine uptake with other transporter substrates. CONCLUSIONS: System ASC/B(0) uptake governs glutamine-dependent growth in colon carcinoma cell lines, and is regulated by a phorbol ester-sensitive pathway that may involve PKC. The results further establish the link between glutamine uptake and colon carcinoma cell growth, a relationship worthy of further investigation with the goal of discovering novel cancer therapeutic targets. PMID- 10792957 TI - Hepatic integrity dependent on matrix metalloproteinase inhibition, not tumor necrosis factor alpha or different bleeding rates. AB - Discrepancies in the levels of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha) following hemorrhagic shock (HS) may be due to the inconsistent rates of bleeding. The purpose of this study was to investigate the effects of rapid versus slow bleeding rates on TNF-alpha levels and if inhibition of TNF-alpha convertase by a matrix metalloproteinase inhibitor (MMPI) affects hepatic integrity in animals exposed to 35% HS. Sprague-Dawley male rats (n = 24, 300-350 g) were divided into four groups: HS 15 (produced over 15 min), HS 30 (produced over 30 min), and HS with MMPI (2.5 mg/kg British Biotech 1101: HS15 + MMPI, HS30 + MMPI). Mean arterial blood pressure (MAP), serum TNF-alpha,levels, and hepatic resting membrane potentials (E(m)) were obtained. A Student t test was performed. TNF-alpha levels for HS 15, HS15 + MMPI, HS 30, and HS 30 + MMPI were 474, 40, 32, and 50 pg/ml, respectively. The hepatic resting membrane potentials for HS 15, HS15 + MMPI, HS 30, and HS 30 + MMPI were -26, -30, -23, and -31 mV, respectively. In conclusion, circulating TNF-alpha levels are affected by the rate of bleeding in hemorrhagic shock. However, despite the differences in the magnitude of TNF-alpha in untreated animals, hepatic integrity was compromised. Interestingly, MMPI, an inhibitor of TNF-alpha convertase, stabilizes the membrane potential in both types of hemorrhagic shock. PMID- 10792958 TI - The efficacy of thyroidectomy for Graves' disease: A meta-analysis. AB - BACKGROUND: Surgery for Graves' disease was largely replaced in the mid-1900s by radioiodine and antithyroid drugs, due to the belief that they were more safe and effective. Since then, thyroid surgery has improved with preoperative drug therapy and modern operative techniques. Recent clinical studies of thyroidectomy for Graves' disease may not reflect outcomes accurately because of small sample size, especially when estimating ideal thyroid remnant size. The purpose of this study was to combine modern clinical trials and use meta-analysis to determine the overall efficacy of both total (TT) and subtotal thyroidectomy (ST) for Graves' disease, compare thyroid function and complications rates of TT and ST, and determine ideal thyroid remnant size. METHODS: Meta-analysis was performed on published studies in which patients underwent either TT or ST for Graves' disease. Meta-analysis was performed by weighted least-squares linear regression. P < 0.05 was considered significant. RESULTS: There were 35 studies comprising 7241 patients. Mean follow-up was 5.6 years. Overall, persistent or recurrent hyperthyroidism occurred in 7.2% of patients. TT was performed on 538 patients and hypothyroidism occurred in all cases. ST was performed in 6703 patients, 59.7% of whom achieved euthyroidism, 25. 6% became hypothyroid, and 7.9% had either persistent or recurrent hyperthyroidism. Permanent recurrent laryngeal nerve injury occurred in 0.9% of TT patients and 0.7% of ST patients (P = NS). Permanent hypoparathyroidism occurred in 1.6% of TT patients and 1.0% of ST patients (P = NS). There was an 8.9% decrease in hypothyroidism and 6.9% increase in euthyroidism for each gram of thyroid remnant (P < 0.0001 each). CONCLUSIONS: Overall, thyroidectomy successfully treated hyperthyroidism in 92% of patients with Graves' disease. There were no cases of hyperthyroidism following TT. ST achieved a euthyroid state in almost 60% of patients with an 8% rate of persistent or recurrent hyperthyroidism. There was no significant difference in complication rates between TT and ST. PMID- 10792959 TI - Kupffer cell inactivation delays repair in a rat model of reversible biliary obstruction. AB - BACKGROUND: During cholestatic liver injury, Kupffer cells (KC) and activated macrophages modulate cell proliferation and subsequent matrix deposition. The role of KC in the restoration of cell architecture and matrix metabolism during repair following chronic cholestatic liver injury is unknown. MATERIALS AND METHODS: To determine the effect of KC inactivation, adult male Sprague-Dawley rats underwent bile duct suspension (BDS) for 5 days followed by reversal of the obstruction. Saline (control) and gadolinium chloride (10 mg/kg) were administered 1 day prior to BDS and 1 day prior to reversal, to inactivate KC during both injury and repair. Serum bilirubin and quantitative cell morphometry were compared to verify the reversibility of the model. Collagen content of the liver was measured in trichrome-stained paraffin sections using NIH imaging software. RESULTS: Reversibility of the obstruction was verified by normalization of direct serum bilirubin, which peaked at 8.42 +/- 0.76 mg/dL following 5 days of BDS and returned to sham-operated levels 2 days after reversal, 0.36 +/- 0.15 mg/dL. Hematoxylin and eosin (H&E)-stained paraffin-embedded liver sections from gadolinium-treated animals at 4 and 7 days after reversal exhibited persistent bile duct proliferation, matrix deposition, and inflammation. Gadolinium-treated animals had altered collagen metabolism compared to saline controls. Whereas the collagen content in the saline group slowly returned to sham-operated levels over time, the treatment group demonstrated progressive accumulation of collagen during repair which was statistically significant at 7 days following reversal (8.79%/mm(2) +/- 2.17 in gadolinium group vs 2. 33%/mm(2) +/- 0.34 in saline group, P = 0.0003). CONCLUSIONS: These results demonstrate that inactivation of resident hepatic macrophages during liver repair impairs collagen metabolism, inhibits the resolution of fibrosis, and allows the persistence of inflammatory cell infiltrates in the portal areas. This is the first evidence of profibrogenic responses in the absence of an intact KC compartment during repair after cholestatic injury. PMID- 10792960 TI - Sublytic complement attack increases intracellular sodium in rat skeletal muscle. AB - BACKGROUND: Although excessive complement activation and deranged sodium homeostasis in skeletal muscle are characteristic in sepsis, their relationship has not been examined. This study was designed to determine if sublytic complement activation can directly mediate changes in myocellular sodium content. MATERIALS AND METHODS: Fast-twitch extensor digitorum longus muscles were freshly isolated from infant rats. Unsensitized muscles were incubated at 30 degrees C for 60 min in the media containing 10% human or rat serum under conditions of no complement activation, activation by zymosan, inactivation by heat, C7 or C9 deficiency, selective inhibition of complement pathway, and inhibition of Na(+) K(+) ATPase by ouabain. Intracellular sodium ([Na(+)](i)) and potassium ([K(+)](i)) contents of the muscles, myocellular ATP, and LDH release from the muscles were then determined. RESULTS: Normal human serum significantly increased [Na(+)](i) and the [Na(+)](i)/[K(+)](i) ratio in the muscles as well as zymosan activated serum. Heat inactivation, C7 deficiency, and inhibition of the alternative pathway completely abolished the cationic changes. Average LDH release was identical in all groups and less than 6%. Complement activation did not impair ouabain-sensitive Na(+)-K(+) ATPase activity in the muscles or alter myocellular ATP. Thus, the observed alterations are not likely due to dysfunction of Na(+)-K(+) pump or depletion of myocellular energy. Instead, alterations in [Na(+)](i) were dependent upon the amount of C9 added to C9-deficient serum, which suggests that the alterations are likely dependent on transmembrane pores created by membrane attack complexes (MAC). CONCLUSIONS: Sublytic amounts of MAC formed as a result of complement activation can directly alter [Na(+)](i) in ex vivo skeletal muscle. PMID- 10792961 TI - All-trans-retinoic acid decreases vein graft intimal hyperplasia and matrix metalloproteinase activity in vivo. AB - BACKGROUND: Development of vein graft intimal hyperplasia has been associated with increased activity of matrix metalloproteinases (MMPs). All-trans-retinoic acid (atRA) decreases expression and activity of MMPs in tissue culture and has decreased intimal hyperplasia following arterial balloon catheter injury. We examined the effect of oral administration of atRA on intimal hyperplasia and MMP expression in an animal model of vein bypass grafting. MATERIALS AND METHODS: Interposition jugular vein bypass grafts were placed in the carotid artery of New Zealand white rabbits. Animals received either atRA (10 mg/kg/day) or vehicle (corn oil) for a period of 2 weeks. Retinoic acid serum levels were determined by HPLC. Intimal and medial areas were measured using morphometric analysis of perfusion-fixed vein graft specimens, and intimal thickness was calculated using circumferential measurements. Expression of MMP-2, MMP-9, and TIMP-1 in vein grafts and unoperated control veins was determined using Northern analysis, and proteolytic activity was determined using substrate gel zymography. RESULTS: Animals treated with atRA had significantly elevated serum levels of this compound and its metabolites. A decrease in intimal to medial ratio was noted after 28 days in vein grafts from treated animals (0.63 vs 0.88, P < 0.01), and a decrease in calculated intimal thickness was noted at 7 and 28 days. Expression of MMP-2 was decreased in treated animals 7 days following surgery, and expression of both MMP-2 and MMP-9 was decreased at 28 days. A decrease in proteolytic activity was noted on zymography at 68 kDa, 7 and 28 days following surgery in vein grafts from animals treated with atRA, corresponding with a decrease in the active form of MMP-2. Increased expression of TIMP-1 was noted in vein grafts from both the treated and the control groups, 7 and 28 days following graft placement. CONCLUSIONS: Oral administration of all-trans-retinoic acid resulted in decreased intimal hyperplasia in an animal model of vein bypass grafting. This was associated with decreased expression and activity of MMP-2 in treated animals. PMID- 10792962 TI - Adenosine preconditioning reduces both pre and postischemic arrhythmias in human myocardium. AB - INTRODUCTION: Consistently, clinical series record supraventricular tachyarrhythmias in approximately 30% of patients following coronary artery bypass surgery (CABG). Ischemic preconditioning and adenosine preconditioning (Ado-PC) decrease postischemia/reperfusion (I/R) myocardial stunning, infarct size, and pharmacologically induced arrhythmias in all species including man. We hypothesized that adenosine preconditioning would decrease spontaneous pre- and postischemic atrial arrhythmias in human myocardium. The purposes of this study were to determine the effect of in vivo and in vitro Ado-PC on atrial arrhythmias. METHODS: Human atrial trabeculae were harvested from CABG patients, placed in organ baths, and paced (1 Hz). Developed force (DF) was recorded during simulated I/R (30/45 min). Prior to I/R, trabeculae were treated with Ado (125 microM) for 5 min (in vitro), or patients were treated with Ado (12 mg iv) 5 min (in vivo) prior to harvest of trabeculae. Contraction frequency >4 Hz (defined as atrial tachyarrhythmias) was recorded in all groups pre- and postischemia. RESULTS: Control trabeculae exhibited increased tachyarrhythmias pre- and postischemia. In vivo and in vitro Ado-PC suppressed both pre- and postischemic arrhythmias. CONCLUSIONS: Adenosine preconditioning suppresses the frequency of pre- and postischemic tachyarrhythmias against an ischemia/reperfusion insult in human myocardium. This antiarrhythmic effect occurs with both in vitro and in vivo administration of adenosine. Preconditioning with adenosine prior to elective ischemia/reperfusion is a promising strategy of reducing spontaneous atrial arrhythmias in patients undergoing myocardial revascularization. PMID- 10792963 TI - Effects of hydrogen peroxide scavenger Catalase on villous microcirculation in the rat small intestine in a model of inflammatory bowel disease. AB - This study was conducted to quantify the effect of systemic Catalase, a hydrogen peroxide scavenger, on villous microcirculation in the inflamed small intestine of the rat. Intestinal inflammation was induced with s.c. application of Indomethacin. Intravital fluorescence microscopy and FITC-labeled erythrocytes were used to quantify erythrocyte velocity and arteriolar diameter in the main arteriole of the villi in the terminal ileum following i.v. application of Catalase in the inflamed intestine, and the blood flow was calculated. Control groups were formed for Ringer's lactate, Catalase and Indomethacin, respectively. We found that villous blood flow was significantly increased in the in the inflamed intestine. Application of Catalase led to a significant decrease in villous perfusion, but had no effect in the control group. The increase in villous blood flow was accompanied by changes in the diameter of the main arteriole. This effect on arteriolar diameter was reversed by i.v. Catalase. Our results provide evidence that systemic application of Indomethacin leads to vasodilatation of the main arteriole of the villus in the rat ileum and hyperemia in the mucosa. Hyperemia and the vascular diameter of the main arteriole were significantly reduced by H(2)O(2)-scavenger Catalase, suggesting that endogenous H(2)O(2) may be one of the mediators of hyperemia in the mucosa in this animal model of intestinal inflammation. PMID- 10792964 TI - Serum activation unmasks unique sensitivities to adhesion molecule expression in human coronary artery and heart microvascular endothelial cells. PMID- 10792965 TI - Concentration dependence of interstitial flow buffering by hyaluronan in synovial joints. AB - Hyaluronan concentration in synovial fluid spans a 20-fold range, from as low as 0.2 mg ml(-1) in synovitis to as high as 4 mg ml(-1) in healthy joints. The aim was to determine the effect of this on fluid drainage from the joint cavity. The study extends the finding of P. J. Coleman, D. Scott, R. M. Mason, and J. R. Levick (1999, J. Physiol. 514, 265-282) that dissolved hyaluronan at 3.6-4.0 mg ml(-1) (the concentration in young human and rabbit joints) causes the opposition to interstitial fluid drainage to increase with pressure. Hyaluronan was infused into rabbit knees at 0, 0.2, 2.0, and 4.0 mg ml(-1) over a range of intraarticular pressures. Hyaluronan at 2 mg ml(-1) (as in healthy elderly joints and some osteoarthritis) greatly reduced drainage rates and generated a flattening (convex) pressure-flow relation, as observed previously with 4 mg ml( 1). Drainage rates were greater at 2 mg ml(-1) than at 4 mg ml(-1) hyaluronan (P < 0.0001, ANOVA, n = 7). The opposition to outflow (pressure required to drive unit outflow) increased with pressure, but less markedly than with 4 mg ml(-1) hyaluronan. Hyaluronan at 0.2 mg ml(-1) reduced outflow by approximately 50% relative to Ringer solution (P < 0.0001, ANOVA, n = 7) but the pressure-flow relation no longer flattened out with increasing pressure, because there was no significant increase in opposition to outflow with pressure. At 0 mg ml(-1) hyaluronan, outflow opposition decreased with pressure. Viscometry showed a marked transition in the hyaluronan state at >/=1.35 mg ml(-1), indicating that this is the critical concentration for molecular domain overlap and intermolecular coupling. The results broadly supported the concentration polarization hypothesis, which predicts significant osmotic buffering of drainage at >/=1 mg ml(-1) hyaluronan; at 0.2 mg ml(-1) other factors may predominate. It is inferred that hyaluronan at physiological concentrations can conserve synovial fluid when pressures are raised (e.g., flexion): whereas dilution of hyaluronan, as in severe effusions, can effectively abolish buffering and thus facilitate fluid drainage. PMID- 10792966 TI - Prostaglandin E(1) is able to increase migration of leukocytes through endothelial cell monolayers. AB - Leukocyte interactions with endothelial cells play an important role during inflammatory processes. Leukocytes pass a monolayer of endothelial cells (ECM) to migrate into the extravascular space. The aim of the current study was to investigate whether prostaglandin E(1) (PGE(1)) influences the process of leukocyte migration. In a prospective controlled study, the influence of prostaglandin E(1) (50-5000 ng/mL) on leukocyte migration through endothelial cell monolayers (n = 7) was investigated. Human umbilical endothelial cells (HUVEC) and/or leukocytes were preincubated with clinically relevant, higher, and lower concentrations of prostaglandin E(1) and the amount of leukocyte migration after 3 h was measured. HUVEC were cultured on microporous membrane filters until achievement of a monolayer for investigation of leukocyte migration. Polymorphonuclear leukocytes (PMNL) were isolated from healthy volunteers and PMNL migration was studied under the influence of PGE(1). In clinically relevant concentrations, PGE(1) was able to increase significantly leukocyte migration through endothelial cell monolayers (205 +/- 7.8%, P < 0.05 compared to control; when treating PMNL alone, migration rate was 120 +/- 9.2% compared to control, ns; only endothelial cell monolayers treated up to 145 +/- 10.2%, P < 0.05 compared to control) showing a dose-dependent effect. In this assay, both cell types (PMNL and ECM) could be treated simultaneously, simulating the clinical situation after an iv administration. In conclusion, PGE(1) is able to increase leukocyte migration through endothelial cell monolayers when both cell types are pretreated. The treatment of either leukocytes or endothelial cell monolayers in the cell coculture showed no significant increase. These findings support the theory that prostaglandins may play a major role during inflammation. Future clinical studies are warranted to confirm this hypothesis. PMID- 10792967 TI - A new model of lung metastasis for intravital studies. AB - We created anexperimental model of pulmonary metastasis based on subcutaneously implanted Lewis lung cancer in mice and observed in vivo the microcirculation of spontaneously metastasized tumors in the lung. The mice lung was held by a small handmade suction ring to stop cardiac and respiratory movement. Using fluorescent microscopy, tumor microcirculation and normal lung microcirculation in the same lung lobe were compared by measuring microvessel diameter and blood flow velocity [red blood cell (rbc) velocity]. In normal microcirculation, the mean values of microvessel diameter and rbc velocity were 10.4 +/- 2.7 microm and 188 +/- 63 microm/s, respectively. In tumor microcirculation, the mean values of the same were 10.6 +/- 3.3 microm and 105 +/- 40 microm/s. The rbc velocity in normal microcirculation was significantly higher (P < 0.001) than that in tumor microcirculation. The calculated shear rates of normal microcirculation and tumor microcirculation were 73.4 +/- 23.4 (/s) and 41.2 +/- 16.1 (/s), respectively. The shear rate of the tumor microcirculation was significantly slower (P < 0.001) than that of the normal microcirculation. We demonstrated a feasibility of observation and measurement of tumor microcirculation in the lung and confirmed that the physiologic data were compatible to those in the brain or in the liver reported by others. This model might be useful for studying metastatic tumor pathophysiology in the lung microcirculation. PMID- 10792968 TI - Quantitative assessment of angiogenesis and tumor vessel architecture by computer assisted digital image analysis: effects of VEGF-toxin conjugate on tumor microvessel density. AB - Tumor growth is angiogenesis dependent. As a consequence, strategies aimed at disrupting this mechanism are heavily investigated. Several angiogenesis assays are used to directly compare the efficacy of anti-angiogenic compounds. However, objective assessment of new vascular growth has been difficult to achieve. The aim of this study was to test and develop a computer-assisted image analysis method that would give an unbiased quantification of the microvessel density. Human tumors were grown in athymic mice and tumor biopsies were taken after a weeklong treatment with VEGF-toxin conjugate. Frozen tumor sections were prepared and stained with PE-conjugated anti-CD-31 antibodies and vessels were imaged with a fluorescence microscope. Vessel density was analyzed by quantifying PE-positive pixels per recorded field. In addition, images were further processed to investigate morphological differences by an automated binarization and skeletonization protocol. This procedure allowed the computer-assisted estimation of important angiogenic parameters such as total vessel number, length, and branch points. Based on these indices, differences in the angiogenic response between control tumors and those treated with VEGF-toxin conjugate were readily detected (P < 0.007 for all parameters). More importantly, computer-generated measurements correlated well with manual microvessel counts and showed significantly less variation. Our results suggest that computer-assisted image analysis represents a rapid, objective, and alternative method for the quantitative assessment of tumor angiogenesis and vessel architecture. PMID- 10792970 TI - Sodium fluorescein-induced changes in leukocyte-endothelial interaction in an animal model. PMID- 10792969 TI - Isolation and culture of capillary endothelial cells from the eel, Anguilla rostrata. AB - There has been little development of endothelial cell cultures from vertebrates other than mammals. In this report the isolation of capillary endothelial cells from the rete mirabile of the eel, Anguilla rostrata, is described. The cells are isolated with collagenase H and dispase II. The cells are plated into fibronectin hyaluronic acid coated flasks. The culture medium is M199 with Earle's salts supplemented with NaCl, HEPES, NaHCO(3), glutamine, pyruvate, heparin, antibiotics, endothelial cell growth supplement, and 20% serum. Cultures are incubated at 25 degrees C in humidified air. The rete mirabile contains pericytes in addition to endothelial cells. Variations in plating time, serum concentrations, and growth matrices were tried to separate the two cell types. The total number of endothelial cells and the ratio of endothelial cells to pericytes are the most important factors in obtaining pure cultures of capillary endothelial cells. Endothelial cells are isolated also from the endocardium, bulbus arteriosus, and large vessels. The initial isolates usually take 3-6 weeks to grow to confluence with subcultures taking about 2 weeks to confluence. PMID- 10792971 TI - Alterations in skeletal muscle microvascular hematocrit with short-term reduced renal mass hypertension. PMID- 10792972 TI - Identification of tumor angiogenesis-related genes by subtractive hybridization. PMID- 10792973 TI - Lateral gene transfer in prokaryotes. PMID- 10792974 TI - Inference from gene trees in a subdivided population. AB - This paper studies gene trees in subdivided populations which are constructed as perfect phylogenies from the pattern of mutations in a sample of DNA sequences and presents a new recursion for the probability distribution of such gene trees. The underlying evolutionary model is the coalescent process in a subdivided population. The infinitely-many-sites model of mutation is assumed. Ancestral inference questions that are discussed are maximum likelihood estimation of migration and mutation rates; detection of population growth by likelihood techniques; determining the distribution of the time to the most recent common ancestor of a sample of sequences; determining the distribution of the age of the mutations on the gene tree; determining in which subpopulation the most recent common ancestor of all the sequences was; determining subpopulation ancestors, where they were, and times to them; and determining in which subpopulations mutations occurred. A computational technique of Griffiths and Tavare used is a computer intensive Markov chain simulation, which simulates gene trees conditional on their topology implied by the mutation pattern in the sample of DNA sequences. The software GENETREE, which implements these ancestral inference techniques, is available. PMID- 10792975 TI - Population outbreaks in a discrete world. AB - We present and analyze a simple three-patch host-parasitoid model where population growth is discrete. The model gives solutions that are qualitatively similar to the stable large-amplitude patterns in space found in reaction diffusion theory. In the context of host-parasitoid interactions, the large amplitude portions of the solution can be thought of as spatially localized host population outbreaks. Here, we show that the biological requirements for localized population outbreaks in a discrete world are identical to those found in reaction- diffusion theory. Furthermore, the model conveniently allows investigation into the robustness of these population outbreaks under the influence of density-dependent dispersal behavior. We find that localized population outbreaks in space can still occur with modest amounts of pursuit and aggregative behavior by parasitoids. We end by showing that evidence from a real host-parasitoid system is consistent with the predictions of the model. PMID- 10792976 TI - Times on trees, and the age of an allele. AB - In this paper we consider the genealogy of a random sample of n chromosomes from a panmictic population which has evolved with constant size N over many generations. We address two related problems. First we describe how genealogical information may be usefully partitioned into information on the events (mutations and coalescences) which occur in the genealogy, and the times between these events. We show that the distribution of the times given information on the events is particularly simple and describe how this can considerably reduce the computational burden when performing inference for these times. Second we investigate the effect on the genealogy of conditioning on a single mutation having occurred during the ancestry of the sample. In particular we use results from the first part of the paper to derive explicit formulae for the density of the age of a mutant allele, conditional on its frequency in either a sample or the population. PMID- 10792977 TI - Evolving populations with overlapping generations. AB - In this paper, we extend a previously published model of an evolving finite population of multi-locus organisms, where the dynamics of the evolving system are described by the cumulants of the population efficacy distribution. We consider the case of overlapping generations and compare it to the previously studied case where generations are discrete. In the weak selection limit, we can solve the dynamics analytically and show that the changes in population genetic variance due to stochastic effects-genetic drift-is twice as great when generations overlap. The comparison of the dynamics of the two models shows many of the features seen in the comparison, performed by Moran, of simple one-locus genetic models with overlapping and non-overlapping generations. Studying the dynamics of the two models gives some insights into these comparisons. PMID- 10792978 TI - Competition by allelopathy proceeds in traveling waves: colicin-immune strain aids colicin-sensitive strain. AB - Producing toxic chemicals to suppress both the growth and survivorship of local competitors is called allelopathy; some strains of the bacteria Escherichia coli produce a toxin (named colicin) which may kill colicin-sensitive neighbors while they themselves are immune. In a previous paper, the competitive outcome between colicin-producing and colicin-sensitive strains was shown to differ between a spatially structured and a completely mixed population. In this paper, we analyze the role of a third, "colicin-immune," strain, which does not produce colicin but is immune to it. Without spatial structure, the colicin-immune strain suppresses the colicin-producing strain and enables the colicin-sensitive strain to win. In a spatially structured population, modeled as a reaction-diffusion system, we examine the speed of boundaries between areas dominated by different strains in traveling waves and the events after the collision of two such boundaries. The colicin-immune strain passes through the area dominated by the colicin-sensitive strain and drives the colicin-producing strain to extinction. Subsequently the colicin-sensitive strain occupies the whole population. PMID- 10792979 TI - Coexistence of macroparasites without direct interactions. AB - Coexistence of macroparasites is studied by extending the infinite-dimensional model considered by Anderson and May (1978, J. Anim. Ecol. 47, 219-247, 249-267) to several species of parasites that are assumed to interact only by causing the death of a common host. An exact invadability condition is found for this model. By studying when mutual invasibility is possible, the region where two parasite species can coexist is found. The result is that, if there is a trade-off between virulence and transmissibility, then coexistence of two species of parasites is possible, but only when the parameters of the model fall into a very narrow parameter region. If, on the other hand, one parasite is more virulent and less transmissible, then it will be competitively excluded. This latter result, though expected in terms of competition theory, is in contrast with what found in the approximate models so far used for studying interacting macroparasites. The effect of parasite aggregation on coexistence is studied by considering two modifications of the basic model (clumped infections and host population heterogeneity in predisposition to infections) that allow for higher aggregation. It appears that the width of the coexistence region is insensitive to these modifications. PMID- 10792980 TI - The number of lines of descent and fixation probabilities of alleles in the pure genetic drift process: analytical approximations. AB - Analysis of diversity between populations diverged from an evolutionarily small number of generations cannot be done under the assumption that allele frequencies reflect an equilibrium between genetic drift and mutations. An alternative to analysis through coalescence theory is proposed in this situation by developing analytical approximations. Using a Poisson approximation of its distribution, the expected number of genes from one generation whose copies make up the population after a number of generations characterized by fixation index F can be shown to be approximated by 2/F-1, irrespective of population size, and probabilities of fixation of alleles over a finite period of time can be also approximated. These expressions, which were checked numerically, should make it possible to calculate approximate likelihoods for allele frequency distributions promoted by drift. PMID- 10792981 TI - The index of dispersion of molecular evolution: slow fluctuations. AB - The most simple neutral model of molecular evolution predicts that the number of substitutions within a lineage in T generations ought to be Poisson distributed. Therefore, the variance in the number of substitutions ought to equal the mean number. The ratio of the variance to the mean number of substitutions is called the index of dispersion, R(T). Assuming infinite sites, no recombination model of the gene, and a haploid, Moran population structure, R(T) is derived for a general stationary model of molecular evolution. R(T) is shown to be affected by fluctuations in parameters only when they occur on a very slow time scale. In order for parameter fluctuations to cause R(T) to deviate significantly from one, the time between parameter changes must be roughly as large, or larger, than the time between substitutions. PMID- 10792982 TI - Viruses at the edge of adaptation. AB - How vulnerable is the line that separates adaptation from extinction? Viruses, in particular RNA viruses, are well known for their high rates of genetic variation and their potential to adapt to environmental modifications (Drake and Holland, 1999; Domingo et al., 2000). Yet, fitness variations-both increases and decreases can be spectacularly rapid, and the simple genetic stratagem of forcing virus multiplication to go through repeated genetic bottlenecks can induce fitness losses, at times near viral extinction. New information has been recently obtained on the two sides of the survival line: the edge of adaptation and the edge of extinction. PMID- 10792984 TI - The Epstein-Barr virus latent membrane protein 1 (LMP1) enhances TNF alpha induced apoptosis of intestine 407 epithelial cells: the role of LMP1 C-terminal activation regions 1 and 2. AB - Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) can protect some kinds of lymphocytes from apoptotic cell death. In contrast, the present study showed that the expression of LMP1 induced high susceptibility to tumor necrosis factor alpha (TNFalpha)-induced apoptosis in intestine 407 epithelial cells, without affecting expression of TNF receptors I and II. LMP1-deletion mutants lacking either C-terminal activation region (CTAR)-1 or CTAR-2 had ability to enhance TNFalpha-induced apoptosis, whereas the deletion of both activation regions completely abolished the induction of high susceptibility to TNFalpha. Phosphorylation of the NFkB-inhibitory molecule IkB-alpha, another biological activity of TNFalpha, was not enhanced by LMP1-expression. LMP1 upregulated antiapoptotic gene A20 expression, suggesting that A20 can not block TNFalpha induced apoptosis in this cell system. Apoptosis triggered by TNFalpha in LMP1 expressing intestine 407 cells was blocked by inhibitors of caspases-8 and -3. It is therefore concluded that in intestine 407 epithelial cells, LMP1 enhances primarily signal cascade responsible for TNFalpha-induced apoptosis, which occurs at a level upstream of acting site of caspases-8 and -3 and that CTAR-1 and CTAR 2 are involved in enhancement of TNFalpha-induced apoptosis. PMID- 10792983 TI - Minor capsid protein of human genital papillomaviruses contains subdominant, cross-neutralizing epitopes. AB - Vaccination with virus-like particles (VLP), comprising both L1 and L2 of human papillomavirus (HPV) genital types 6, 16, and 18, induces predominantly type specific neutralizing antibodies. L2 polypeptide vaccines protect animals against experimental challenge with homologous papillomavirus and cross-reactive epitopes are present in HPV L2. To assess L2-specific cross-neutralization of HPV genotypes, sheep were immunized with purified, bacterially expressed HPV6, 16, or 18 L2. In addition to neutralizing the homologous HPV type in vitro, antisera to each HPV L2 also cross-neutralized both heterologous HPV types. This suggests that unlike VLP-based prophylactic HPV vaccines, an L2 polypeptide vaccine may provide broad-spectrum protection. PMID- 10792985 TI - Independent introduction of transmissible F/D recombinant HIV-1 from Africa into Belgium and The Netherlands. AB - Most HIV-1 subtype F viruses described so far have been isolated from individuals originating in South America, Romania, or Central Africa. Previous studies have shown that subtype F viruses from these three areas can be distinguished by phylogenetic tree analysis of various parts of the HIV genome. Subtype F strains circulating in Central Africa and classified as subgroup F2 and F3 have relatively large nucleotide distances from strains of subgroup F1, which includes some African strains, along with strains from Romania and South America. Subtype F strains have now appeared in Europe. In this study, we analyzed the complete gag gene and a large fragment of the pol gene of seven strains of African origin that represent the three F subgroups. At least five of the seven strains appear to be intersubtype recombinants. Of four strains circulating in Belgium and the Netherlands, three were F/D mosaics and the fourth harboured a G(gag)/GH(pol)/F3(env) recombinant structure. Two of the three F/D mosaics showed identical breakpoints and were independently introduced in Belgium and the Netherlands. At least two of the mosaics were further transmitted. The remaining three strains of the seven we studied were isolated from individuals in Cameroon. Two included large or smaller F1 fragments in gag and pol. The third strain was subtype D along the entire gag and pol fragment. A parental African subtype F that showed no evidence for recombination was not found. PMID- 10792986 TI - Varicella-zoster virus infection of a human CD4-positive T-cell line. AB - Varicella-zoster virus (VZV) is a human alpha-herpesvirus that causes varicella (chickenpox) at primary infection and may reactivate as herpes zoster. VZV is a T lymphotropic virus in vivo. To investigate the T-cell tropism of VZV, we constructed a recombinant virus expressing green fluorescent protein (VZV-GFP) under the CMV IE promoter. Coculture of VZV-GFP-infected fibroblasts with II-23 cells, a CD4-positive human T-cell hybridoma, resulted in transfer of virus to II 23 cells. II-23 cells are susceptible to VZV-GFP infection as demonstrated by expression of immediate/early (IE62), early (ORF4), and late (gE) genes. Recovery of infectious virus was limited, with only 1 to 3 in 10(6) cells releasing infectious virus by plaque assay, indicating that transfer of virus results in a limited productive infection. In vitro infection of II-23 cells will be useful for further analysis of VZV tropism for T-lymphocytes. PMID- 10792987 TI - Efficient complementation by chimeric Microviridae internal scaffolding proteins is a function of the COOH-terminus of the encoded protein. AB - Microviridae morphogenesis is dependent on two scaffolding proteins, an internal and external species. Both structural and genetic analyses suggest that the COOH terminus of the internal protein is critical for coat protein recognition and specificity. To test this hypothesis, chimeric internal scaffolding genes between Microviridae members phiX174, G4, and alpha3 were constructed and the proteins expressed in vivo. All of the chimeric proteins were functional in complementation assays. However, the efficient complementation was observed only when the viral coat protein and COOH-terminus of internal scaffolding were of the same origin. Genes with 5' deletions of the phiX174 internal scaffolding gene were also constructed and expressed in vivo. Proteins lacking the first 10 amino acids, which self-associate across the twofold axes of symmetry in the atomic structure, efficiently complement phiX174 am(B) mutants at temperatures above 24 degrees C. These results suggest that internal scaffolding protein self associations across the twofold axes of symmetry are required only at lower temperatures. PMID- 10792988 TI - HTLV-1 tax oncoprotein binds to DNA topoisomerase I and inhibits its catalytic activity. AB - HTLV-1 Tax oncoprotein exerts pleiotropic effects on cellular regulatory systems, such as transcription and the cell cycle, through the interaction with various cellular factors. During our search for additional cellular targets of Tax using a yeast two-hybrid screening system, we isolated a cDNA encoding human DNA topoisomerase I. Tax was demonstrated to bind to topoisomerase I in vitro, and the Tax-topoisomerase I complex was also detected in HTLV-1-infected T-cells by immunoprecipitation. Furthermore, Tax inhibited the catalytic activity of topoisomerase I as measured by relaxation of supercoiled DNA, although complete inhibition was not observed under the conditions used. The binding of topoisomerase I to DNA was inhibited by the addition of the wild type of Tax but not by a mutant of Tax that cannot bind to topoisomerase I. Consistent with these observations, expression of Tax induced an in vivo reduction of the covalent association of topoisomerase I with chromosomal DNA, which accumulates in the presence of camptothecin. These results suggest that Tax has a novel potential to affect various cellular processes such as transcription and maintenance of genomic stability, in which DNA topoisomerase I is involved. PMID- 10792989 TI - Human interleukin-6 facilitates hepatitis B virus infection in vitro and in vivo. AB - BACKGROUND AND AIM: Research on hepatitis B virus (HBV) infection in vivo has been limited due to the absence of a suitable animal model. We have developed a human-mouse radiation chimera in which normal mice, preconditioned by lethal total body irradiation and radioprotected with SCID mouse bone marrow cells, are permissive for engraftment of human hematopoietic cells and solid tissues. This resulting human-mouse model, which comprises three genetically disparate sources of tissue, is therefore termed Trimera. This study was aimed at assessing the effect of human IL-6 on HBV infection in vivo in Trimera mice. METHODS: Trimera mice were transplanted with human liver tissue fragments or with HepG2-derived cell lines, which had been previously infected ex vivo with HBV in the presence or absence of human interleukin-6 (hIL-6) and in the presence of anti-IL-6 neutralizing antibodies. RESULTS: HBV sequences appeared in the sera of animals in which the liver tissue was incubated with both HBV and hIL-6 prior to transplantation. A similar result was obtained when a human hepatoblastoma cell line (HepG2), expressing the hIL-6 receptor, was infected ex vivo with HBV in the presence of hIL-6 prior to their injection into spleens of Trimera mice. However, when liver fragments were infected ex vivo and simultaneously treated with neutralizing antibodies against hIL-6 or were incubated with HBV prior to transplantation without hIL-6, the rate of mice positive for HBV DNA in their sera was lower. Human mononuclear cells are also permissive for HBV infection in vitro: in the presence of hIL-6 the infection of these cells is enhanced; and this infection is suppressed by the chimeric protein named Hyper-IL-6, generated by the fusion of hIL-6 to the soluble hIL-6 receptor (sIL-6Ralpha, gp80). CONCLUSION: hIL-6 facilitates HBV infection in vitro and in vivo. PMID- 10792990 TI - Multidrug resistance genotypes (insertions in the beta3-beta4 finger subdomain and MDR mutations) of HIV-1 reverse transcriptase from extensively treated patients: incidence and association with other resistance mutations. AB - Multiple nucleoside resistance involves specific mutational patterns of the HIV-1 pol gene that are independent of the classic mutations conferring resistance to individual dideoxynucleosides. These include a cluster of five mutations in the reverse-transcriptase (RT) coding region (A62V, V75I, F77L, F116Y, and Q151M) generally referred to as multidrug resistance (MDR) mutations, and insertions of one or several amino acid residues between codons 67 and 70 of RT, a flexible region joining two antiparrallel beta sheets (beta3-beta4 insertions). The objectives of this study were (i) to determine the prevalence of multidrug resistance genotypes (MDR mutations and beta3-beta4 insertions) in a cohort of 632 patients who were extensively pretreated with anti-HIV drugs and not responding to their current antiretroviral therapy, and (ii) to analyze the association of multidrug resistance genotypes with other resistance mutations in the RT and protease genes. Among viruses sequenced from these patients, 15 (2.4%) of them contained an insertion and 2 (0.3%) contained a deletion in the beta3 beta4 finger subdomain of RT. In 9 cases, the insertion was associated with a D67S, G, or E mutation. In addition, we identified 13 (2.1%) viruses harboring specific MDR mutations (mainly Q151M and/or A62V, V75I, F116Y). Interestingly, the A62V mutation was found in 6 of the 15 strains with an insertion, whereas the other MDR mutations were not observed in insertion mutant strains. Especially high levels of resistance to zidovudine were observed for viruses with a beta3 beta4 insertion in the background of A62V, L210W, and T215Y. Otherwise, MDR mutations and beta3-beta4 insertions were found in association with the classic mutations conferring resistance to zidovudine, lamivudine, nonnucleoside RT inhibitors, and protease inhibitors, according to treatment history. Finally, we observed a genome with a deletion of codon 70 associated with a Q151M MDR mutation. These data suggest that the emergence of HIV-1 multidrug resistance, which may occur in various genetic contexts, poses a challenging problem in formulating treatment strategies. PMID- 10792991 TI - Characterization of HIV-1-specific cytotoxic T lymphocytes expressing the mucosal lymphocyte integrin CD103 in rectal and duodenal lymphoid tissue of HIV-1 infected subjects. AB - Acute HIV-1 infection depletes CD4(+) T cells in gut-associated lymphoid tissue (GALT). The failure of containment of local viral replication, and consequent CD4(+) T cell depletion, might be due to delayed mobilization of effector CD8(+) T cells or absence of functioning HIV-1-specific CD8(+) T cell effectors within GALT. No studies have addressed human intestinal HIV-1-specific CD8(+) T cell functions. We sought to determine whether functional HIV-1-specific CTL were present in GALT and whether the repertoire differed from HIV-1-specific CTL isolated from peripheral blood mononuclear cells. From three HIV-1-infected subjects, we isolated HIV-1-specific CD8(+) T cells expressing the mucosal lymphocyte integrin CD103 from GALT. These antigen-specific effector cells could be expanded in vitro and lysed target cells in an MHC class I-restricted manner. HIV-1-specific CTL could be isolated from both duodenal and rectal GALT sites, indicating that CD8(+) effectors were widespread through GALT tissue. The breadth and antigenic specificities of GALT CTL appeared to differ from those in peripheral blood in some cases. In summary, we found HIV-1-specific CD8(+) effector T cells in GALT, despite HIV-1-induced CD4(+) T cell lymphopenia. This suggests that HIV-1-specific CTL in gut tissue can be maintained with limited CD4(+) T cell help. PMID- 10792992 TI - Sequences flanking the cAMP responsive core of the HTLV-I tax response elements influence CREB protease sensitivity. AB - The human T cell leukemia virus type I (HTLV-I) Tax protein activates transcription from the viral long terminal repeat and select cellular promoters by interacting with cellular DNA-binding proteins. The HTLV-I promoter contains three copies of a Tax-responsive element (TRE-1), each of which possesses a core cAMP response element (CRE). The cAMP response element-binding protein, CREB, binds TRE-1 and mediates Tax association with, and transactivation of, the viral promoter. These activities depend on DNA sequences that flank the core CRE. Although CREs are found in a variety of cellular promoters, cellular CREs vary in sequence from TRE-1, especially in the flanking regions, and are generally not Tax responsive. The molecular basis for differential Tax responsiveness of viral and cellular CREs has not been determined. Here we demonstrate that the conformation of CREB is influenced by the nucleotide sequence of its DNA-binding element. CREB showed altered sensitivity to V8, chymotrypsin, and trypsin proteases when bound to the HTLV-I TRE-1 element as compared to the rat somatostatin CRE element. The phosphorylation state of CREB did not influence its protease sensitivity on either element. Sequences flanking the core CRE-binding site in each element were found to specify protease sensitivity. Since the TRE-1 flanking sequences also modulate Tax association with CREB, and Tax transactivation of CREB-dependent LTR transcription, these results suggest that CREB conformation may determine the ability of Tax to bind CREB. PMID- 10792993 TI - HIV-1 replication is inhibited by a pseudo-substrate peptide that blocks Tat transactivation. AB - The activation of the HIV-1 long terminal repeat (LTR) by the viral transcriptional transactivator Tat is an essential step in the viral replication cycle. To increase the processivity of RNA polymerase II, Tat interacts with the positive transcription elongation factor b (P-TEFb) and cyclin-dependent kinase (CDK)-activating kinase (CAK). In this study, we demonstrate that a pseudo substrate peptide for CDK7, mC2p, inhibits HIV-1 replication as well as Tat transactivation. Specifically, mC2p blocks only the activity of CAK and not that of P-TEFb. Moreover, mC2p inhibits Tat transactivation and HIV replication. Therefore, the activation of CDK7 by Tat is considered a critical step of Tat transactivation and mC2p and related compounds represent potential candidates for novel anti-HIV therapeutics. PMID- 10792995 TI - Hepatitis B virus assembly is sensitive to changes in the cytosolic S loop of the envelope proteins. AB - Among the three related L, M, and S envelope proteins of the hepatitis B virus (HBV), the L and S polypeptides are required for virion production. Whereas the pivotal function of the pre-S region of L in nucleocapsid envelopment has been established, the contribution of its S domain and the S protein is less clear. In this study, we evaluated the role of the cytosolic S loop, common to L and S, in HBV assembly by performing mutagenesis experiments. To distinguish between the effect of the mutations on either envelope or virion formation, we investigated the ability of the mutants to assemble into secretable subviral empty envelopes and to replace the wild-type proteins in virion maturation, respectively. Virion production was found to be blocked by each of the secretion-competent deletion and substitution mutants SDelta35-39, SDelta40-46, SDelta50-56, and Svarsigma56 59, while an insertion within the loop is tolerated. Surprisingly, single mutations of the arginines terminating the loop had an opposite effect: while a conservative exchange of Arg-73 still allowed virion formation, the same mutation of Arg-79 did not. The critical sequences and/or structural requirements of the cytosolic S loop involved in nucleocapsid envelopment primarily act in the S background. These findings can be related to a model for a synergistical function of both L and S proteins in HBV morphogenesis. PMID- 10792994 TI - The E1 helicase of human papillomavirus type 11 binds to the origin of replication with low sequence specificity. AB - Expression of the human papillomavirus type 11 E1 and E2 genes is necessary and sufficient to support viral DNA replication. The full-length E2 protein is a transcriptional modulator that also interacts with the E1 helicase to form an E1/E2 complex at the viral origin of replication. Previous studies indicated that efficient binding of this complex to the replication origin is site-specific and that the E2 homodimer was required for efficient E1 binding. Human papillomavirus type 11 E2 and E1 proteins have been purified and their cooperative binding to the HPV type 11 viral replication origin has been characterized. Low-affinity E1 binding to the HPV type 11 replication origin was demonstrated and found to be largely nonspecific. DNA binding by E1 does not require complex formation with E2 and appears to be independent of ATP binding or hydrolysis. E1 binding quantitatively increased with the addition of increasing amounts of E2 and mutations in the E2 binding site demonstrated that the E2BS site is required for E1 and E2 to specifically bind as a high-affinity complex at the replication origin. Analysis of the A/T-rich E1 binding site via mutation showed that it was nonessential for high-affinity E1/E2 complex formation. Thus, although the replication functions between the animal and the human papillomaviruses are well conserved, there are subtle differences in the DNA binding requirements for E1, which may portend mechanistic differences among the DNA replication systems of various papillomavirus types. PMID- 10792996 TI - Oligomerization, secretion, and biological function of an anchor-free parainfluenza virus type 2 (PI2) fusion protein. AB - A number of studies indicate that the transmembrane domain, the cytoplasmic domain, or both regions of viral surface glycoproteins are involved in quaternary structure formation. In this report, the transmembrane domain and cytoplasmic tail coding sequence of the fusion (F) glycoprotein gene from parainfluenza type 2 virus was truncated by PCR and the resulting gene (PI2F') was expressed in HeLa T4 cells by using the vaccinia virus-T7 transient expression system. Pulse-chase experiments indicated that the anchor-free PI2F' was expressed and processed into F(1) and F(2) subunits. Both the processed and the unprocessed anchor-free PI2F' proteins were found to be efficiently secreted into the culture medium. Examination of the oligomeric form of the anchor-free PI2F' by chemical cross linking demonstrated that it assembles posttranslationally into dimers and trimers with a pattern similar to that of the wild-type PI2F protein. In an effort to better understand the biological properties of the truncated form of PI2F', we anchored PI2F' by a glycosyl-phosphatidylinositol (GPI) linkage. The GPI-anchored PI2F' protein, when coexpressed with PI2HN, did not induce cell fusion seen as syncytium formation, but was found to initiate lipid mixing (hemifusion) as observed by transfer of R-18 rhodamine from red blood cells to the GPI-PI2F'/PI2HN cotransfected cells. The results therefore indicate that the extracellular domain of the PI2 fusion protein contains not only the structural information sufficient to direct assembly into higher oligomers, but also is competent to initiate membrane fusion, suggesting that the anchor-free PI2F' may be useful for further structural studies. PMID- 10792997 TI - Molecular studies on bromovirus capsid protein. AB - Specific interactions are likely to occur between the highly conserved N-proximal arginine-rich motif (ARM) of Brome mosaic virus (BMV) coat protein (CP) and each of three genomic RNAs and a single subgenomic RNA during in vivo encapsidation. To characterize these interactions, three independent deletions were engineered into a biologically active clone of BMV RNA3 (B3) such that the matured CP of each B3 variant precisely lacks either the entire ARM (B3/Delta919) or two consecutive arginine residues (B3/13DeltaDelta14 and B3/18DeltaDelta19) within the ARM. Analysis of virion RNA for each B3 variant recovered from symptomatic leaves of Chenopodium quinoa revealed that the interactions between the N terminal ARM of BMV CP and each of three genomic RNAs is distinct. Northern blot hybridization of B3Delta919 virion RNA revealed that the deleted ARM region specifically affected the stability of virions containing RNA1. An abundant truncated RNA species recurrently found in the virions of B3Delta919 was identified to be a derivative of genomic RNA1, lacking the 5' 943 nucleotides. Additional Northern blot analysis of virion RNAs from B3/Delta919, B3/13DeltaDelta14, and B3/18DeltaDelta19, and in vitro reassembly assays revealed that the N-terminal ARM region contains crucial amino acids required for RNA4 packaging, independent of genomic RNA3. The significance of these observations in relation to Bromovirus CP-RNA interactions during virion assembly is discussed. PMID- 10792998 TI - Synergistic interactions of antibodies in rate of virus neutralization. AB - Antibodies and antibody combinations are often evaluated only by their potency in inactivating a known quantity of virus in dose-effect assays. However, a crucial additional parameter is the rate at which neutralization takes place, or kinetics. Synergism of certain antibody combinations in dose-effect assays has been previously demonstrated. In the present report, using a battery of murine monoclonal antibodies to herpes simplex virus (HSV), we investigated whether antiviral antibodies can also synergize in neutralization kinetics. To determine whether synergism in dose-effect assays can predict synergism in neutralization rate, the ability of neutralizing antibodies to synergize in neutralization rate (kinetics) was compared to their ability to synergize in dose-effect assays (potency) in cell-free assays. Although certain antibody combinations synergized in both neutralization rate and potency, combinations that did not clearly synergize in potency could still significantly synergize in neutralization rate. Weak neutralizing antibodies could also greatly increase the neutralization rate of more potent antibodies. These results suggest that evaluating antibody combinations in dose-effect assays but not in neutralization kinetics provides a partial picture of neutralizing antibody dynamic interactions and may prevent the identification of certain favorable antibody combinations. These findings also support the importance of establishing defined antibody cocktails for prophylactic and therapeutic purposes. A simple strategy to evaluate antibody interactions in neutralization kinetics is proposed in which a quantitative prediction of additivity is made on the basis of the neutralization rate constants of the individual antibodies in the combination. PMID- 10792999 TI - Retinoic acid resistance at late stages of human papillomavirus type 16-mediated transformation of human keratinocytes arises despite intact retinoid signaling and is due to a loss of sensitivity to transforming growth factor-beta. AB - In our in vitro model of human cell carcinogenesis, normal human foreskin keratinocytes (HKc) transfected with human papillomavirus type 16 DNA (HKc/HPV16) progress toward malignancy through several phenotypically defined and reproducible "steps" that include immortalization, growth factor independence (HKc/GFI), differentiation resistance (HKc/DR), and ultimately malignant conversion. While HKc/HPV16 are very sensitive to growth inhibition by all-trans retinoic acid (RA) at early passages, they lose their sensitivity to RA during progression in culture. However, gel mobility shift assays using the retinoid response elements DR1 and DR5 showed no changes in binding activity of nuclear extracts obtained from HKc/HPV16 at different stages of in vitro progression. Similarly, Western blot analyses for retinoic acid receptor gamma-1 and the retinoid X receptors failed to reveal any decreases in the levels of these retinoid receptors throughout progression. In addition, luciferase activity driven by the SV40 promoter with a DR5 enhancer element was activated following RA treatment of HKc/DR that were resistant to growth inhibition by RA. Since RA induces transforming growth factor-beta2 (TGF-beta2) in normal HKc and HKc/HPV16, we investigated whether this response changed during progression. Again, RA induced TGF-beta2 mRNA in early and late passage HKc/HPV16, HKc/GFI, and HKc/DR approximately to the same extent, confirming that the RA signaling pathways remained intact during in vitro progression despite the fact that the cells become resistant to growth inhibition by RA. We then investigated the sensitivity of HKc/HPV16 to growth inhibition by TGF-beta. While early passage HKc/HPV16 were as sensitive as normal HKc to growth inhibition by TGF-beta1 and TGF-beta2, the cells became increasingly resistant to both TGF-beta isotypes during in vitro progression. In addition, while both RA and TGF-beta produced a decrease in the levels of mRNA for the HPV16 oncogenes E6 and E7 in early passage HKc/HPV16, this effect was also lost at later stages of progression. Finally, blocking anti-TGF beta antibodies partially prevented RA inhibition of growth and E6/E7 expression in early passage HKc/HPV16. Taken together, these data strongly suggest that inhibition of growth and HPV16 early gene expression in HKc/HPV16 by RA is mediated by TGF-beta and that a loss of RA sensitivity is linked to TGF-beta resistance rather than alterations in RA signaling. PMID- 10793000 TI - Loss of transforming growth factor-beta (TGF-beta) receptor type I mediates TGF beta resistance in human papillomavirus type 16-transformed human keratinocytes at late stages of in vitro progression. AB - Human keratinocytes (HKc) immortalized by human papillomavirus type 16 DNA (HKc/HPV16) progress toward malignancy through growth factor-independent (HKc/GFI) and differentiation-resistant stages (HKc/DR). This progression is associated with a loss of sensitivity to growth inhibition by both all-trans retinoic acid (RA) and transforming growth factor-beta (TGF-beta). In the accompanying article (Borger et al., 2000, Virology 270, 397-407), we demonstrate that RA resistance in HKc/HPV16 arises despite functional nuclear retinoid receptors and that TGF-beta mediates growth inhibition by RA. To investigate the basis for the loss of TGF-beta sensitivity during in vitro progression of HKc/HPV16, we explored the expression of TGF-beta receptors type I and type II in independently derived HKc/HPV16 lines and their corresponding HKc/GFI and HKc/DR derivatives. While TGF-beta receptor type II mRNA levels were unchanged during progression, mRNA levels for TGF-beta receptor type I decreased dramatically as the cells became TGF-beta resistant. At the HKc/DR stage, loss of TGF-beta receptor type I mRNA, compared to low-passage cells, ranged from 55 to 87% in four HKc/HPV16 lines examined. Immunohistochemistry, using anti-TGF-beta receptor type I antibodies, confirmed a loss of TGF-beta receptor type I expression in HKc/DR. Reintroduction of the TGF-beta-receptor type I into TGF-beta-resistant HKc/DR completely restored growth inhibition by TGF-beta. Southern blot analysis of DNA extracted from normal HKc, HKc/HPV16, and HKc/DR ruled out any gross changes in the TGF-beta receptor type I gene. The activity of the TGF-beta receptor type I promoter, cloned upstream of a luciferase reporter gene, was decreased in HKc/DR, to an extent comparable to the decrease in mRNA levels for the TGF-beta receptor type I. Thus, TGF-beta resistance at late stages of HPV16 mediated transformation of HKc is the result of a loss of expression of TGF-beta receptor type I. PMID- 10793001 TI - Identification and phylogenetic comparison of Salem virus, a novel paramyxovirus of horses. AB - A virus that could not be identified as a previously known equine virus was isolated from the mononuclear cells of a horse. Electron microscopy revealed enveloped virions with nucleocapsid structures characteristic of viruses in the Paramyxoviridae family. The virus failed to hemabsorb chicken or guinea pig red blood cells and lacked neuraminidase activity. Two viral genes were isolated from a cDNA expression library. Multiple sequence alignments of one gene indicated an average identity of 45% as compared to Morbillivirus N protein sequences. A weaker relationship was found with Tupaia paramyxovirus (TPMV) and Hendra virus (HeV) N proteins. In the second gene, multiple open reading frames (ORFs) were identified, corresponding to the arrangement of the P, V, and C ORFs in the Morbillivirus and Respirovirus viruses. Short stretches in the C-terminal regions of the P and C proteins showed limited homologies to viruses in the Morbillivirus genus but no obvious relationship to viruses in other genera. The V ORF translation product contained a highly conserved, cysteine-rich domain that is common to most viruses in the Paramyxovirinae subfamily. Sequencing of P gene cDNA clones confirmed the use of a cotranscriptional editing mechanism for the regulation of P/V expression. Based on the location of its origin it has been named Salem virus (SalV). PMID- 10793002 TI - The bovine papillomavirus E2 transactivator is stimulated by the E1 initiator through the E2 activation domain. AB - Bovine papillomavirus type 1 (BPV-1) encodes two regulatory proteins, E1 and E2, that are essential for viral replication and transcription. E1, an ATP-dependent helicase, binds to the viral ori and is essential for viral replication, while the viral transcriptional activator, E2, plays cis-dominant roles in both viral replication and transcription. At low reporter concentrations, E1 stimulates E2 enhancer function, while at high reporter concentrations, repression results. An analysis of cis requirements revealed that neither replication nor specific E1 binding sites are required for the initiators' effect on E2 transactivator function. Though no dependence on E1-binding sites was found, analysis of E1 DNA binding and ATPase mutants revealed that both domains are required for E1 modulation of E2. Through the use of E2 fusion-gene constructs we showed that a heterologous DNA-binding domain could be substituted for the E2 DNA-binding domain and this recombinant protein remained responsive to E1. Furthermore, E1 could rescue activation domain mutants of E2 defective for transactivation. These data suggest that E1 stimulation of E2 involves interactions between E1 and the E2 activation domain on DNA. We speculate that E1 may allosterically interact with the E2 activation domain, perhaps stabilizing a particular structure, which increases the enhancer function of E2. PMID- 10793003 TI - Rotavirus VP6 expressed by PVX vectors in Nicotiana benthamiana coats PVX rods and also assembles into viruslike particles. AB - The rotavirus major inner capsid protein (VP6) has been expressed in Nicotiana benthamiana plants using vectors based on potato virus X (PVX). VP6 was expressed either as a fusion with the PVX coat protein or from an additional subgenomic promoter inserted to enable both VP6 and PVX coat protein to be expressed independently. Both approaches yielded VP6, which retained the ability to form trimers. VP6 expressed from the subgenomic promoter assembled into paracrystalline sheets and tubes. Expression as a fusion protein yielded PVX rods that presented an external "overcoat" of VP6, but unexpectedly, some rotavirus protein also assembled into icosahedral viruslike particles (VLPs). The assembly of viral protein into VLPs suggests that prior display of VP6 on the flexuous PVX rod facilitates the subsequent assembly of VP6 into stable icosahedral particles. PMID- 10793004 TI - Immune protection against HSV-2 in B-cell-deficient mice. AB - The role of antibody in protection of the vaginal mucosa and sensory ganglia against HSV-2 infection was examined using HSV- immune, B-cell-deficient muMT mice. Significantly higher virus titers were detected in the vaginal mucosae of immune muMT mice compared to immune C57BL/6J mice 24 h after HSV-2 rechallenge. However, virus was rapidly cleared in immune muMT mice, and the infection was resolved with only a 2-day delay. Passive transfer of immune serum to immune muMT mice prior to rechallenge resulted in HSV-specific vaginal IgG levels comparable to those of immune C57BL/6J mice. Although transferred antibody failed to prevent reinfection of the majority of recipients, vaginal virus titers at 24 h and clearance kinetics were similar to those of immune C57BL/6J controls. Following vaginal rechallenge, HSV-2 did not spread to the sensory ganglia of immune C57BL/6J mice nor was the rechallenge virus detected in the ganglia of the majority of immune muMT mice. However, protection was severely compromised by T cell depletion of immune C57BL/6J mice. These results suggest that HSV-specific antibody limits, but does not prevent, infection of the genital epithelia. Further, prevention of virus spread to the sensory ganglia in immune animals requires vigorous T-cell immune responses. PMID- 10793005 TI - Structure-function analysis of the Sendai virus F and HN cytoplasmic domain: different role for the two proteins in the production of virus particle. AB - The role of the cytoplasmic domain (cytd) of the Sendai virus HN and F glycoproteins in the process of virus assembly and budding are evaluated. Recombinant Sendai virus (rSeV) mutants are generated carrying modifications in the cytd of each of the glycoprotein separately. The modifications include increasing truncations and/or amino acid sequence substitutions. Following steady state (35)[S]methionine/cysteine labeling of the infected cells, the virus particle production is estimated. The radioactive virions in the cell supernatants are measured relative to the extent of the infection, assessed by the intracellular N protein signal. For both the F and HN cytd truncation mutants, the largest cytd deletions lead to a 20- to 50-fold reduction in virion production. This reduction cannot be explained by a reduction of the cell surface expression of the glycoproteins. For the F protein mutants, the virions produced in reduced amount always exhibit a normal F protein composition. It is then concluded that a threshold level of F is required for SeV assembly and budding. The rate or the efficiency with which this threshold is reached up appears to depend on the nature of the F cytd. A minimal cytd length is required as well as a specific sequence. The analysis of HN protein mutants brings to light an apparent paradox. The larger cytd truncations result in significant reduction of virion production. On the other hand, a normal virion production can take place with an underrepresentation of or, even, an undetectable HN in the particles. The HN uptake in virion is confirmed to depend on the previously proposed cytd SYWST signal (T. Takimoto, T. Bousse, E. C. Coronel, R. A Scroggs, and A. Portner. 1998. J. Virol. 72, 9747-9754.). PMID- 10793006 TI - Cleavage of hepatitis C virus nonstructural protein 5A by a caspase-like protease(s) in mammalian cells. AB - Nonstructural 5A protein (NS5A) of hepatitis C virus (HCV) is localized in the cytoplasm although it has a functional nuclear localization signal. To clarify the determinant of NS5A cytoplasmic localization, various N- or C-terminal deleted NS5A mutants were generated and their subcellular localization was analyzed in cell lines after transient expression. N-terminal deleted forms of NS5A were localized in the nucleus, and the sequence of the N-terminal 27 amino acids of NS5A had sufficient function to cause retention of a normally nuclear protein in the cytoplasm. These observations indicated that cytoplasmic localization of NS5A is determined primarily by the N-terminal region of the molecule. In addition, we found proteolytic processing of NS5A in transiently expressing cells. In these cells, cleavage occurred at a few sites located in the N- and C-terminal regions of NS5A. This cleavage in cells was enhanced by apoptotic stimuli and was inhibited by the caspase inhibitor Z-VAD-FMK, suggesting that a caspase-like protease(s) contributes to the cleavages of NS5A. Based on the results of mutational analysis of NS5A, we predicted one cleaved form, which had lost both the N- and the C-terminal portions of NS5A, to be composed of amino acid residues 155 to 389. Peptide containing the same amino acid sequence as this cleaved product was localized in the nucleus. Furthermore, we found that a fusion protein consisting of Gal4 DNA-binding domain fused with this cleaved form showed transcriptional activity only when the alpha-catalytic subunit of protein kinase A (PKA) was coproduced, suggesting that the transcriptional activity of this product was regulated by PKA. These results suggested that the cleavage product of NS5A by a caspase-like protease(s) plays a role in transcriptional regulation of the host cell gene(s) in HCV-infected cells. PMID- 10793007 TI - A gene for hypotrichosis simplex of the scalp maps to chromosome 6p21.3. AB - Hypotrichosis simplex of the scalp (HSS) is an autosomal dominant form of isolated alopecia causing almost complete loss of scalp hair, with onset in childhood. After exclusion of candidate regions previously associated with hair loss disorders, we performed a genomewide linkage analysis in two Danish families and localized the gene to chromosome 6p21.3. This was confirmed in a Spanish family, with a total LOD score of 11.97 for marker D6S1701 in all families. The combined haplotype data identify a critical interval of 14.9 cM between markers D6S276 and D6S1607. Localization of the locus for HSS to 6p21.3 is a first step toward identification of the gene. The gene will give important insights into the molecular and cellular basis of hair growth on the scalp. PMID- 10793008 TI - Phenotypes of patients with "simple" Mendelian disorders are complex traits: thresholds, modifiers, and systems dynamics. PMID- 10793010 TI - Cocaine and the kidney: a synthesis of pathophysiologic and clinical perspectives. AB - Cocaine abuse has reached epidemic proportions in the United States, and several forms of renal disease have been associated with this widespread use. The hemodynamic actions of cocaine, as well as its effects on matrix synthesis, glomerular inflammation, and glomerulosclerosis, may contribute to renal injury. Cocaine abuse has been associated with various forms of acute renal failure and acid-base and/or electrolyte disorders and may also have a role in the progression of chronic renal failure to end-stage renal disease. In utero exposure to cocaine has been associated with urogenital tract anomalies. Medical management of a hypertensive emergency caused by acute cocaine toxicity requires a multisystem approach, with close monitoring of cardiac, neurological, and renal functions. PMID- 10793009 TI - Genomewide search for type 2 diabetes susceptibility genes in four American populations. AB - Type 2 diabetes is a serious, genetically influenced disease for which no fully effective treatments are available. Identification of biochemical or regulatory pathways involved in the disease syndrome could lead to innovative therapeutic interventions. One way to identify such pathways is the genetic analysis of families with multiple affected members where disease predisposing genes are likely to be segregating. We undertook a genomewide screen (389-395 microsatellite markers) in samples of 835 white, 591 Mexican American, 229 black, and 128 Japanese American individuals collected as part of the American Diabetes Association's GENNID study. Multipoint nonparametric linkage analyses were performed with diabetes, and diabetes or impaired glucose homeostasis (IH). Linkage to diabetes or IH was detected near markers D5S1404 (map position 77 cM, LOD = 2.80), D12S853 (map position 82 cM, LOD = 2.81) and GATA172D05 (X chromosome map position 130 cM, LOD = 2.99) in whites, near marker D3S2432 (map position 51 cM, LOD = 3.91) in Mexican Americans, and near marker D10S1412 (map position 14 cM, LOD = 2.39) in African Americans mainly collected in phase 1 of the study. Further analyses showed evidence for interactions between the chromosome 5 locus and region on chromosome 12 containing the MODY 3 gene (map position 132 cM) and between the X-chromosome locus and region near D12S853 (map position 82 cM) in whites. Although these results were not replicated in samples collected in phase 2 of the GENNID study, the region on chromosome 12 was replicated in samples from whites described by Bektas et al. (1999). PMID- 10793011 TI - Antibodies to topical bovine thrombin correlate with access thrombosis. AB - Bovine thrombin is often used topically to promote hemostasis during vascular surgery, including dialysis-access placement. Patients frequently develop antibodies to bovine thrombin preparations, and some may develop antiphospholipid antibodies. We evaluated 88 hemodialysis patients for the presence of antibodies to topical bovine thrombin to determine if elevated antibody levels correlated with vascular access thrombosis. Twenty-seven patients (30.7%) had elevated antibody levels to topical bovine thrombin. More patients with elevated antibody levels had prior vascular access thrombosis than patients with normal antibody levels (13 of 27 versus 5 of 61 patients; P < 0.001). This difference was almost entirely the result of greater levels of thrombosis in patients with polytetrafluoroethylene (PTFE) grafts and elevated antibody levels. In these patients, 11 of 13 patients (84.6%) with elevated antibody levels had a previous thrombosis compared with 2 of 15 patients (13. 3%) with normal antibody levels (P < 0.001). Patients with elevated antibody levels and PTFE grafts also had more prior thromboses (1.92 +/- 1.60 versus 0.133 +/- 0.35 thromboses; P < 0.01) and a greater thrombosis rate (66.89 +/- 63.71 versus 4.65 +/- 12.05 thromboses/100 patient-years; P < 0.01) than patients with normal antibody levels. There were no differences in the frequency of myocardial infarction, coronary artery bypass, access age, presence of diabetes mellitus, platelet counts, anticardiolipin antibody, albumin, lactate dehydrogenase, or C-reactive protein levels. In conclusion, patients with PTFE grafts and elevated antibody levels to topical bovine thrombin had significantly more vascular access thrombosis. PMID- 10793012 TI - Regional citrate anticoagulation in continuous venovenous hemodiafiltration. AB - Over the past several years, continuous venovenous hemodiafiltration (CVVHDF) using pump-driven devices has gained wide acceptance as a form of renal replacement therapy for critically ill patients with acute renal failure. More recently, regional citrate anticoagulation has proven useful as a method of anticoagulating CVVHDF circuits, particularly in those patients at high risk for bleeding. However, an easy and convenient method for guiding the dose of citrate infusion has not previously been described. We describe the use of an algorithm using posthemofilter levels of ionized calcium to guide the dose of administered regional citrate on the survival time and urea and creatinine clearances of 24 Hospal AN69HF hemofilters. Nine patients with acute and chronic renal failure requiring CVVHDF were studied. The median filter survival time when using the postfilter ionized calcium algorithm was 3.4 days, with a survival probability of 46% (95% confidence interval [CI], 17 to 71). Random-effects linear regression analysis did not show a significant decline in blood-side urea clearance (P = 0.041) or creatinine clearance (P = 0. 308). Moreover, definite bleeding complications occurred with an incidence rate of 0.045/person-day on citrate anticoagulation (95% CI, 0.006 to 0.16), and occult bleeding occurred with an incidence rate of 0.091/person-day on citrate anticoagulation (95% CI, 0.03 to 0.23). Guiding regional citrate anticoagulation through the use of posthemofilter ionized calcium levels is a safe and effective method of prolonging filter life during CVVHDF. PMID- 10793013 TI - Seasonal variations of blood pressure and overhydration in patients on chronic hemodialysis. AB - Blood pressure (BP) has a seasonal cycle in the general population and in patients undergoing maintenance dialysis, but the causes remain unclear. We studied the BP measurements recorded at fixed hours three times weekly from 1994 to 1997 in 102 hemodialysis patients. We obtained monthly averages of the following variables: predialysis mean BP, greatest overhydration (OH) estimated by predialysis body weight excess over dry weight, chronic OH estimated by the remaining postdialysis weight excess over dry weight, urea reduction ratio (URR) in dialysis, and monthly means for daylight span and outdoor temperature over the study period. Average BP in the population diminished over the 48-month period, associated with a decrease in chronic OH (r = 0.66; P < 0.0005) but independent of greatest OH. BP and chronic OH presented synchronous seasonal variations, with peaks in late autumn and early winter and troughs in summer. These biological rhythms were inversely related to the seasonal daylight span and outdoor temperature. Both BP and chronic OH periods were synchronous with the daylight annual cycle and preceded the seasonal variations of temperature by 1 month. Multiple regression analysis showed that chronic OH and daylight, but not URR or temperature, had a significant independent association with BP changes. These results show the existence of seasonal variations of BP in dialysis patients that are associated and synchronous with seasonal changes in chronic OH status. Both cycles depend on conditions influenced by the annual daylight span more than by external temperature. PMID- 10793014 TI - Cardiac and hemodynamic effects of hemodialysis and ultrafiltration. AB - Imbalance between cardiac oxygen supply and demand may trigger cardiac events in already vulnerable hemodialysis (HD) patients. We studied the effect of ultrafiltration (UF) and HD in nine chronic HD patients by continuously measuring blood volume (BV; by Critline), blood pressure (BP; by Portapres), and changes in hemodynamics (Modelflow) during isolated UF (iUF) of 500 mL in 30 minutes and subsequent HD combined with UF (HD + UF). Aortic pressure was reconstructed from finger pressure. Changes in cardiac oxygen supply were assessed by calculating the area under the aortic pressure curve during diastole (diastolic pressure time index [DPTI]). Changes in cardiac oxygen demand were assessed by calculating systolic pressure time index (SPTI). BV decreased 4.0% +/- 1.8% during UF and 7.3% +/- 3.3% during HD + UF (both P < 0.01). Systolic BP did not change; diastolic and mean BP increased 11 +/- 7.4 and 11 +/- 8.4 mm Hg during iUF, respectively (both P < 0.01), and stabilized during HD + UF. Overall pulse pressure decreased 19 +/- 11.1 mm Hg (P < 0.01). Heart rate increased 13 +/- 11 beats/min (P < 0.01) and systemic vascular resistance increased 59% +/- 51% (P < 0. 01), whereas stroke volume and cardiac output (CO) decreased by 40% +/- 17% and 30% +/- 13%, respectively (both P < 0.01). Both cardiac oxygen supply (DPTI) and demand (SPTI) increased during iUF, and both decreased during HD + UF. By the end of the procedure, DPTI/SPTI ratio had increased 9% +/- 8% (P < 0.05). Changes in CO correlated closely to changes in BV. Despite large changes in hemodynamics during uncomplicated UF and HD, the balance between cardiac oxygen supply and demand (DPTI/SPTI ratio) did not decrease, but improved slightly. PMID- 10793015 TI - Ondansetron therapy for uremic pruritus in hemodialysis patients. AB - Pruritus is a distressing symptom affecting up to 90% of dialysis patients. Conventional treatment with antihistamines is often ineffective and may have unacceptable side effects. Serotonin (5-hydroxytryptamine type 3 [5-HT(3)]) is known to enhance pain perception and pruritic symptoms through receptors on sensory nerve endings. Antagonism of 5-HT(3) receptors may be of use in treating uremic pruritus. We randomly assigned 16 hemodialysis patients with persistent pruritus to treatment with the 5-HT(3)-receptor antagonist, ondansetron (8 mg), or placebo three times daily for 2 weeks each in a prospective, placebo controlled, double-blind crossover study. Patients scored their intensity of pruritus daily on a 0-to-10 visual analogue scale (0 = no pruritus, 10 = maximal pruritus), and daily use of antihistamines as escape medication was recorded. The median daily pruritus score did not change significantly during active or placebo treatment (preondansetron, 5. 3; interquartile range [IQR], 3.4 to 6.3; during ondansetron, 3.9; IQR, 2.7 to 5.0; P = not significant; preplacebo, 3.7; IQR, 3.0 to 4. 6; during placebo, 3.6; IQR, 2.4 to 4.8; P = not significant). The median daily percentage of escape medication use decreased from 21% (IQR, 9 to 61) to 9% (IQR, 0 to 33) with ondansetron (P = not significant) and from 53% (IQR, 0 to 88) to 5% (IQR, 0 to 31) with placebo (P = not significant). There was no difference in predialysis biochemistry test results or dialysis efficacy during treatment phases. Ondansetron does not improve pruritus in hemodialysis patients. Use of antihistamines decreased with both ondansetron and placebo. PMID- 10793017 TI - Effects of dialyzer reuse on the permeability of low-flux membranes. AB - Little attention has been given to the effects of reuse on the permeability of low-flux membranes, especially regarding middle molecules. We studied two different types of low-flux membranes at reuses 0, 6, and 12 in five patients undergoing hemodialysis with the following combinations of membrane and sterilant: cellulose diacetate membrane and formaldehyde, polysulfone membrane and formaldehyde, cellulose diacetate membrane and peracetic acid, and polysulfone and peracetic acid. The permeability of the membranes was assessed through the hydraulic ultrafiltration coefficient (K(UF)), sieving coefficient for beta(2)-microglobulin (B2M), and vitamin B(12) and albumin concentrations in ultrafiltrate. After 12 reuses, total cell volume (TCV) tended to be reduced in both cellulose diacetate and polysulfone dialyzers irrespective of the sterilant used, but significance was only found for the first set of dialyzers. Cellulose diacetate dialyzers reprocessed with either formaldehyde or peracetic acid showed an important reduction in K(UF) (31% [P < 0.05] and 23% [P < 0.05], respectively). A significant elevation in K(UF) was found in polysulfone membranes reprocessed with peracetic acid (41%; P < 0.05), but no alterations in K(UF) were found in polysulfone membranes reprocessed with formaldehyde. Cellulose diacetate membranes were intrinsically more permeable to B2M than polysulfone membranes (sieving coefficient, 6. 85 +/- 2.53 versus 0.04 +/- 0.02 x 10(-2); P < 0.001), which was not modified by any of the sterilants. Vitamin B(12) levels in ultrafiltrate decreased to an undetectable level in four of five samples collected after 12 reuses in polysulfone membranes reprocessed with peracetic acid (90 +/- 71 to 3 +/- 8 pg/mL; P < 0. 05 versus reuse 0). Albumin leakage occurred in two of five samples after the 12th reuse, but only in polysulfone membranes reprocessed with peracetic acid. Our findings suggest that reuse of low-flux polysulfone dialyzers reprocessed with peracetic acid is associated with structural damage of the membrane and a reduced permeability to middle molecules. PMID- 10793016 TI - Serotyping strip immunoblot assay for assessing hepatitis C virus strains in dialysis patients. AB - Recent accumulated evidence shows that dialysis patients are a high-risk group for hepatitis C virus (HCV) infection. Assessment of HCV genotype distribution among dialysis patients may be important because specific viral genotypes are associated with different clinical manifestations, disease progression, and response to antiviral therapy. However, polymerase chain reaction-based methods are cumbersome and unsuitable for analyzing large cohorts of dialysis patients with HCV. Instead, this information can be obtained by using a novel recombinant immunoblot assay (RIBA) recently developed for determining HCV serotype. The RIBA HCV serotyping strip immunoblot assay (SIA; Chiron Corporation, Emeryville, CA), is based on an immunoblot strip with five lanes of immobilized serotype-specific HCV peptides from the nonstructural (NS4) and core regions of the genomes of HCV types 1, 2, and 3. HCV serotype is deduced by determining the greatest intensity of reactivity to the NS4 serotype-specific HCV peptide band in relation to the internal control band (human immunoglobulin G) intensity on each strip. HCV core peptide reactivity is used only in the absence of NS4 reactivity. We compared RIBA HCV serotyping SIA with genotyping using sera from a large (n = 107) cohort of HCV-infected patients undergoing chronic hemodialysis (HD). We successfully serotyped 79 of 107 patients (74%) undergoing HD. We found a remarkable concordance (65 of 70 results; 93%) between RIBA HCV serotyping SIA and genotyping (line probe assay [LiPA]) techniques (kappa = 0.786) with sera from viremic patients infected with a known genotype. Only 5 of 70 patients (7%) had apparently discordant results. In a subset of patients (28 of 107 patients; 26%) not typed by RIBA HCV serotyping SIA, most (24 of 28 patients; 86%) were successfully genotyped by LiPA technology. It was possible to assess serotype reactivity in some patients (9 of 107 patients; 7%) who could not be genotyped. The distribution of HCV serotypes was associated with the antibody response against HCV proteins and the patterns of reactivity by RIBA HCV 2.0 SIA. In conclusion, (1) we found good agreement between serotyping and genotyping methods in our large cohort of dialysis patients infected with HCV; (2) the impaired immunocompetence conferred by uremia may limit serotyping analysis in some HCV infected patients undergoing HD; (3) RIBA HCV serotyping SIA may be useful in tracking transmission routes for HD patients who cleared the virus and have only anti-HCV antibody; and (4) the distribution of HCV serotypes was associated with the antibody response against HCV proteins and the patterns of reactivity by RIBA HCV 2.0 SIA. Assessment of HCV strains appears to be very useful in the routine clinical activity of nephrologists within HD units because consistent biological differences among HCV strains exist. RIBA serotyping SIA is a simple, inexpensive, and highly reproducible assay to obtain information about HCV types in the HD setting. PMID- 10793018 TI - Sexual dysfunction after renal replacement therapy. AB - The existence of a sexual problem as the subjective evaluation of sexual function was assessed with a simple questionnaire. Those questioned were patients undergoing dialysis treatment (n = 400) or with a functioning renal transplant (RTx; n = 300) and both men and women in the general Dutch population (n = 591). In the Dutch control population, 8.7% of the men and 14.9% of the women reported a sexual problem, showing a significant gender difference but unrelated to age. In patients, the prevalence of a sexual problem was significantly greater (hemodialysis, men, 62.9%; women, 75.0%; peritoneal dialysis, men, 69.8%; women, 66.7%; renal transplantation, men, 48.3%; women, 44.4%). In RTx recipients, sexual problems were significantly less prevalent than in patients undergoing dialysis (P < 0.001). Only in male patients was an association between prevalence of a sexual problem and age found. The results of the simple questionnaire were sufficiently validated when 102 of 104 patients confirmed their responses in a subsequent structured interview. This study shows that the prevalence of sexual problems in patients undergoing renal replacement therapy is high and clinically relevant. PMID- 10793019 TI - Atherogenic lipoprotein phenotype in end-stage renal failure: origin and extent of small dense low-density lipoprotein formation. AB - End-stage renal failure (ESRF) is associated with dyslipidemia and accelerated atherosclerosis. Triglyceride-rich lipoproteins accumulate and qualitative changes take place in low-density lipoprotein (LDL), with a predominance of the small dense LDL phenotype. Increased small dense LDL (LDLIII) is a known risk factor for cardiovascular disease. To assess the extent of LDLIII formation in ESRF and identify factors contributing to LDLIII production, we analyzed LDL subfractions by density-gradient ultracentrifugation, very low-density lipoprotein subfractions, and lipase activity in 75 patients with ESRF (25 hemodialysis [HD], 25 peritoneal dialysis [PD], and 25 predialysis patients) and 40 age- and sex-matched controls. The percentage of LDLIII was increased in all three patient groups compared with controls (PD, 33% +/- 29% [mean +/- SD]; P < 0.005; HD, 30% +/- 22%; P < 0.01; predialysis, 26% +/- 26%; P < 0.01; all versus controls, 14% +/- 10%). Plasma LDLIII concentration was increased only in PD patients (median, 84 mg/dL; interquartile range [IQR], 29 to 160 mg/dL versus controls; median, 31 mg/dL; IQR, 26 to 54 mg/dL). In other patient groups, total LDL level was less, with heterogeneity in LDLIII concentrations. Forty percent of PD patients and 28% of HD and predialysis patients had LDLIII concentrations greater than 100 mg/dL compared with 2.5% of controls (P = 0.002). Plasma triglyceride levels (r(2) = 38.4%; P < 0.001) and hepatic lipase activity (r(2) = 6.7%; P < 0.03) were independent predictors of LDLIII concentration. The strong association between LDLIII concentration and triglyceride level was present in all three patient groups (HD, r(2) = 47.9%; PD, r(2) = 45. 2%; predialysis, r(2) = 25.8%); plasma triglyceride levels greater than 177 mg/dL (2.0 mmol/L) had an 86% specificity and 79% sensitivity for predicting an LDLIII concentration greater than 100 mg/dL. We conclude that the atherogenic lipoprotein phenotype predominates in ESRF, with excess LDLIII particularly prominent in PD patients. Atherogenic levels of LDLIII are found in patients with triglyceride levels greater than 177 mg/dL. This is likely to represent a further cardiovascular risk factor in this population. PMID- 10793020 TI - Tubular injury as a cardinal pathologic feature in human heme oxygenase-1 deficiency. AB - Heme oxygenase (HO) catalyzes degradation of heme to biliverdin, iron, and carbon monoxide. It consists of three isoforms: an inducible form (HO-1), a constitutive form (HO-2), and the third isoform (HO-3), with properties similar to HO-2. There is limited evidence to suggest that the induction of HO-1 may have anti inflammatory effects in an in vivo model of oxidative stress-mediated renal injury. We experienced the first human case of HO-1 deficiency. The patient had persistent proteinuria and hematuria, with biochemical evidence of renal tubular injury. We obtained three consecutive renal specimens: two from renal biopsies at 2 and 5 years of age and the third from autopsy at 6 years of age. The patient had systemic vascular endothelial-cell injury with massive intravascular hemolysis. The serum was loaded with heme and a large amount of heme-conjugated haptoglobin. A high concentration of haptoglobin was also detectable in urine. Mesangial proliferation or change in glomerular capillary-wall thickness was relatively mild to moderate in all specimens. Electron microscopic examination showed widespread endothelial detachment and subendothelial deposits of an unidentifiable material. It was striking that tubulointerstitial injury, with tubular dilatation and/or atrophy, interstitial fibrosis, and inflammatory cell infiltration, advanced progressively. Tubular epithelial cells were injured, and massive deposition of iron and haptoglobin was detectable. Bowman's capsules were dilated significantly, probably secondary to the collapse of atrophic tubuli. This is the first report to show that HO-1 has critical roles in vivo in protecting renal tubuli, in addition to vascular endothelium, from oxidative injury. PMID- 10793021 TI - Possible molecular basis for changes in potassium handling in acute renal failure. AB - Renal potassium excretion is diminished in acute renal failure (ARF). The gastrointestinal tract can compensate for deficient renal potassium excretion in many patients with ARF. For both impaired renal potassium excretion and gastrointestinal compensation in ARF, little is known about the role of potassium channels. We hypothesized that specific changes in the expression of the secretory renal outer medullary potassium channel (ROMK), and the potassium channel regulator, channel-inducing factor (CHIF), in kidney and colon could contribute to changes in potassium handling. Sprague-Dawley rats underwent uninephrectomy and 35 minutes of renal ischemia, followed by varying periods of reperfusion. Renal function, serum and urine potassium levels, and aldosterone levels were measured. The expression of messenger RNA (mRNA) for ROMK and CHIF in the kidney and CHIF in the colon were measured by Northern blot hybridization. Serum creatinine level was increased at 24 hours and started to decline at 48 hours of renal ischemia reperfusion injury (IRI). Serum potassium level was increased at 24 hours, further elevated at 48 hours of IRI, and returned to normal at 7 days of IRI. Urine potassium level was reduced at 24 and 48 hours. Northern blot analysis indicated that the expression of ROMK1 mRNA in the cortex or medulla remained unchanged at 24 hours but profoundly decreased (by 70% to 80%) at 48 hours (n = 4; P < 0.01). The expression of CHIF mRNA in the kidney cortex or medulla decreased by 25% to 30% at 24 hours and 35% to 40% at 48 hours of IRI (n = 4; P < 0.05 for each group). CHIF mRNA expression in the distal colon was moderately increased at 24 hours (approximately twofold) and significantly enhanced at 48 hours (more than threefold; P < 0.01; n = 4) of IRI. Serum aldosterone level was increased approximately threefold at 48 hours of IRI (P < 0.01; n = 6). In conclusion, (1) suppression of ROMK and CHIF in the kidney may contribute to decreased renal potassium excretion in ARF; (2) enhanced expression of CHIF in the distal colon in IRI could be an adaptive response to increase potassium excretion in the colon and help modify hyperkalemia in ARF; and (3) increased aldosterone levels, as a response to hyperkalemia, could be upregulating colonic CHIF. PMID- 10793022 TI - Changing incidence of glomerular diseases in adults. AB - Studies performed at large metropolitan medical centers have reported an increasing incidence of idiopathic focal segmental glomerulosclerosis (FSGS) in adults. To determine whether a similar trend occurs in small urban and rural communities and to determine the role of race in these observations, we reviewed the patient records of all adults who underwent renal biopsies at our institution over the 20-year period from 1974 to 1994. The patients were grouped for analysis in 5-year intervals, 1975 to 1979, 1980 to 1984, 1985 to 1989, and 1990 to 1994, for the following diagnoses: FSGS, membranous nephropathy (MN), minimal change nephropathy (MCN), membranoproliferative glomerulonephritis (MPGN), immunoglobulin A (IgA) nephropathy, chronic glomerulonephritis, diabetic nephropathy, hypertensive nephrosclerosis, and chronic interstitial nephritis. Patients with secondary causes for these lesions were excluded. The relative frequency of FSGS increased from 13.7% during 1975 to 1979 to 25% during 1990 to 1994 (P < 0.05). The relative frequency of MN decreased from 38.3% during 1975 to 1979 to 14.5% during 1990 to 1994 (P < 0.01). There were no changes in the frequencies of MCN, MPGN, IgA nephropathy, chronic glomerulonephritis, diabetic nephropathy, hypertensive nephrosclerosis, or chronic interstitial nephritis over the 20-year period. However, there was a significant increase in the percentage of blacks with FSGS, from 0% in 1975 to 1979 to 22.6% in 1990 to 1994, and an increased percentage of Hispanics with FSGS, from 0% in 1975 to 1979 to 21.3% in 1990 to 1994 (P < 0.05). The modest increase in whites with FSGS did not reach statistical significance. The incidence of MN in blacks and whites decreased over the 20-year period. In the last 5 years, 15 patients per year had FSGS compared with 7 patients per year with MN (P < 0.05). No changes in age or sex between groups or over time accounted for these results. We conclude that FSGS is now diagnosed twice as often as MN and is the most common idiopathic glomerular disease at our hospital. Reasons for this increase include the emergence of FSGS in both Hispanics and blacks, with a modest increase of FSGS in whites. The increase in FSGS in the three most common races in our community suggests that factors other than genetic, perhaps environmental, have a role in the pathogenesis of FSGS. PMID- 10793023 TI - Prevalence of HIV-associated nephropathy in autopsies of HIV-infected patients. AB - Previous studies have reported that approximately 10% of the patients with human immunodeficiency virus (HIV) infection develop HIV-associated nephropathy (HIVAN). However, over the last decade, morbidity and mortality as a result of HIV-1 infection has remarkably decreased with the availability of potent new antiretroviral drugs. We therefore determined the prevalence of HIVAN from autopsy data of HIV-infected patients in more recent years (1992 to 1997). Autopsy reports of 389 patients were reviewed. In reports suggestive of possible HIVAN, slides of renal tissue were retrieved and reviewed again to ensure appropriate classification. The criteria for the diagnosis of HIVAN were focal segmental glomerulosclerosis with collapse of the glomerular tuft in some glomeruli, extensive tubular ectasia, and significant tubulointerstitial disease. Of 389 autopsy reports, 54% of the patients were black, 35% were white, and 11% were Hispanic. Thirty-three percent of the patients had a history of intravenous drug abuse. The mean CD4 count of the patients was 54 +/- 91/microL (mean +/- SD). In 27 cases, typical features of HIVAN were found based on the criteria used, accounting for an overall HIVAN prevalence of 6.9% (27 of 389 autopsies). Because the overwhelming majority of these patients were black (93%), the prevalence in blacks was 12% (25 of 209 autopsies). We conclude that although mortality and morbidity from HIV infection is decreasing, HIVAN remains an important complication of HIV infection in blacks, even in recent years. PMID- 10793024 TI - Significantly high regional morbidity of MPO-ANCA-related angitis and/or nephritis with respiratory tract involvement after the 1995 great earthquake in Kobe (Japan). AB - Within a 3-year period after the Great Earthquake of Kobe (Japan) resulted in more than 6,000 deaths and complete destruction of the central area of Kobe City, 14 patients (group 1 [G1]) with myeloperoxidase (MPO)-antineutrophil cytoplasmic autoantibody (ANCA)-related angitis and/or nephritis presented to Nishi-Kobe Medical Center in western Kobe City. On the other hand, only 15 patients with this disease were encountered between 1990 and 1997 at Kyoto University Hospital in Kyoto City, which is located 80 km from Kobe City and was only minimally affected by the earthquake. These 15 patients and 1 patient who presented to Nishi-Kobe Medical Center before the Great Earthquake were classified as group 2 (G2). Although the average MPO-ANCA titer in G1 was almost the same as that in G2, G1 showed a significantly greater average value for white blood cells than G2 (11,321 +/- 4,369 versus 8,116 +/- 2, 389/microL; P < 0.05). Concerning renal function, a significant elevation in creatinine (Cr) levels at diagnosis (7.4 +/- 3.8 versus 2.1 +/- 1.4 mg/dL; P < 0.01) and rapidly declining rates of reciprocal Cr levels were noted in G1 (0.325 +/- 0.304 versus 0.087 +/- 0.069 dL/mg. wk; P < 0.01). The number of patients who required emergency hemodialysis was significantly greater in G1 than G2 (nine versus three patients; P < 0.02); however, the incidence of renal death and mortality were not significantly different between the groups. The number of patients who reported upper respiratory tract inflammation as an initial symptom was also significantly greater in G1 than G2 (eight versus two patients; P < 0.01). Moreover, patients in G1 experienced a significantly greater rate of severe pulmonary involvement during the hospital course than G2 (pulmonary hemorrhage, five versus no patients; interstitial pneumonitis, four versus two patients, respectively; P < 0.01). The relatively uniform and distinctive clinical features of the disease after the Great Earthquake, in conjunction with a high morbidity, suggest a relationship between disease development and this urban type of earthquake. Severely provoking air pollution caused by massive destruction and reconstruction of the city may have caused high frequencies of upper respiratory tract inflammation as an initial symptom and severe pulmonary involvement. PMID- 10793025 TI - Insertion/deletion polymorphism in ACE gene is not associated with renal progression in Japanese patients with IgA nephropathy. AB - We determined the relationship between the gene polymorphism of angiotensin I converting enzyme (ACE) and the progression of immunoglobulin A (IgA) nephropathy in a large cohort in a multicenter trial of ethnically homogeneous Japanese patients (n = 527). Patients with biopsy-proven IgA nephropathy were recruited from several clinics in Japan. The mean observation period was 8.4 +/- 4.7 years. ACE insertion/deletion (I/D) genotype was determined by polymerase chain reaction amplification using allele-specific primers. Clinical factors investigated in all patients were date of birth, sex, levels of urinary protein excretion, duration of observation, serum creatinine (sCr) level, and creatinine clearance (CCr). ACE genotype distribution did not differ between patients who maintained normal renal function (II, 41%; ID, 44.7%; DD, 14.3%) and those who progressed to renal impairment (II, 41.7%; ID, 40.4%; DD, 17.9%). Kaplan-Meier analysis did not show a significant difference in renal survival rate among the three groups of each genotype. In multivariate analysis, only two variables, proteinuria greater than 1.0 g/d of protein and impaired renal function (sCr >1.2 mg/dL or CCr <70 mL/min) at the time of renal biopsy, were found to be risk factors for disease progression leading to a poor outcome. No association was observed between these variables and ACE genotype. It appears that ACE I/D polymorphism may not affect the progressive deterioration of renal function in patients with IgA nephropathy from our multicenter trial. PMID- 10793026 TI - Factors predictive of outcome in severe lupus nephritis. Lupus Nephritis Collaborative Study Group. AB - In 1992, we published the results of a prospective, controlled trial of aggressive therapy (high-dose prednisone plus oral cyclophosphamide alone or with plasmapheresis) in 86 patients with severe lupus nephritis. During this study, remission (serum creatinine < or =1.4 mg/dL [< or =123 micromol/L] and proteinuria < or =330 mg/d of protein) in renal disease occurred in 37 patients (43%). To assess the long-term effect of remission on patient and renal survival, we now report the results of our extended follow-up of these patients. After an average of 10 years of follow-up in the 86 patients, patient survival rates at both 5 and 10 years were 95% in the group that had a remission and 69% at 5 years and 60% at 10 years in the no-remission group (P < 0.001). Renal survival rates were 94% at both 5 and 10 years in the remission group compared with 46% at 5 years and 31% at 10 years in the no-remission group (P < 0. 0001). Features predictive of remission included stable renal function after 4 weeks on therapy, category IV lesion, lower chronicity index, white race, lower urine protein excretion level at baseline, and lower baseline serum creatinine level. The features predictive of end-stage renal disease were higher baseline serum creatinine level, presence of anti-Ro antibodies, and failure to attain a remission. Thus, in patients with the most severe forms of lupus nephritis, a remission of clinical renal abnormalities is associated with dramatic improvement in long-term patient and renal survival. PMID- 10793027 TI - Access to renal transplantation among patients with end-stage renal disease due to lupus nephritis. AB - Access to living related renal and cadaveric renal transplantation was compared between 5,863 adults with incident end-stage renal disease (ESRD) caused by lupus nephritis and 463,141 adults with other causes of ESRD who were included in the US Renal Data System from 1987 to 1995. Using Cox regression models that adjusted for differences in age, sex, race, region of residence, and year of onset of ESRD, patients with ESRD caused by lupus nephritis were as likely as patients with other causes of ESRD to receive a living related renal transplant (adjusted hazard ratio [HR] = 1.02; 95% confidence interval [CI], 0.93 to 1.10; P = 0.70) but were 20% less likely to receive a cadaveric renal transplant (adjusted HR = 0.80; 95% CI, 0.75 to 0.85; P < 0.0001). Patients with ESRD caused by lupus nephritis were significantly more likely to be entered onto a waiting list for cadaveric renal transplantation (adjusted HR = 1. 15; 95% CI, 1.10 to 1.21; P < 0.0001) but were less likely to receive a cadaveric transplant once entered onto a waiting list (adjusted HR = 0.73; 95% CI, 0.69 to 0.78; P < 0.0001). Patients with ESRD caused by lupus nephritis had equal access to living related renal transplantation and greater enrollment on waiting lists for cadaveric transplantation than patients with ESRD from other causes, indicating that medical ineligibility is not a major barrier to transplantation. Both medical and nonmedical factors may contribute to the decreased likelihood of cadaveric transplantation among patients with ESRD caused by lupus nephritis. PMID- 10793028 TI - Impaired adaptation to renal mass reduction in the polycystic rat. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a serious cause of renal failure. In many renal-disease models, surgical renal mass reduction accelerates disease progression. We explored whether surgical renal mass reduction and the method of renal mass reduction accelerate the course of ADPKD. Studies were conducted in male heterozygous cystic Han:SPRD rats and unaffected littermate controls. Control and cystic rats were subjected to 50% renal mass reduction by uninephrectomy, 50% renal mass reduction by infarction of half of each kidney, or sham operation. Most groups were followed up to the age of 20 weeks, with serial measurements of blood pressure and proteinuria. At 20 weeks, glomerular filtration rate (GFR) and renal plasma flow (RPF) rate were measured. Similar studies to 12 weeks of age were performed in additional groups of control and cystic rats with either sham operation or 50% renal infarction. In noncystic rats, uninephrectomy led to minimal effects on blood pressure and proteinuria and to substantial compensatory renal hypertrophy, hyperfiltration, and hyperperfusion. Similar renal mass reduction by segmental infarction led to greater values for blood pressure and proteinuria and significant compensatory hyperfiltration. In contrast, the cystic rats showed a significant reduction in baseline renal blood flow, more profound increases in blood pressure and proteinuria, and no compensatory increases in GFR and RPF after reduction of renal mass. These studies suggest that the ability of cystic kidneys to respond to acquired loss of nephrons is impaired and that these kidneys are at greater risk when additional renal injury is superimposed. PMID- 10793029 TI - Gender-dependent effect of L-NAME on polycystic kidney disease in Han:SPRD rats. AB - To determine whether the renal nitric oxide (NO) system has a role in the pathogenesis of polycystic kidney disease (PKD) in Han:Sprague-Dawley (SPRD) rats, the NO synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L NAME), 70 mg/L, or L-arginine, 0.5 g/L, was administered to heterozygous diseased (cy/+) and homozygous normal animals. Urine nitrate and nitrite excretion was reduced by L-NAME treatment and, in the male cy/+ rats, increased by L-arginine administration. The administration of L-NAME significantly increased blood pressure in all groups, whereas L-arginine had no effect. L-NAME and L-arginine had a modest but significant overall effect on the severity of cystic disease in male rats, reflected by relative kidney weights and cyst volume densities. This effect was gender dependent because it was not observed in female animals. The administration of L-NAME resulted in a significant increase in plasma creatinine concentration of the cy/+ rats, which was more marked in male than female animals. These observations support the recently reported gender differences in the renal NO system and a small role for NO synthesis that can be inhibited by L NAME in the pathogenesis of PKD in Han:SPRD rats. These observations do not exclude a more important role for the endogenous renal NO production in the pathogenesis of PKD in view of a recent report of a major NOS resistant to conventional inhibitors in the rat kidney. PMID- 10793031 TI - Renal tubular acidosis and vasculitis associated with IgE deposits in the kidney and small vessels. AB - We report a woman with a history of allergies, polyuria, polydipsia, proteinuria, renal loss of electrolytes, renal tubular acidosis, nephrocalcinosis, and palpable purpura. A proximal defect was excluded by a normal bicarbonate reabsorption curve, and a distal tubular defect was shown because urine pH did not decrease to less than 6.4 despite ammonium chloride-induced systemic acidosis. Moreover, furosemide failed to improve urinary acidification. Urine-to blood PCO(2) gradient was less than 14 mm Hg, although the urine bicarbonate level reached values as high as 89 mEq/L. Combining bicarbonate and neutral phosphate infusions increased the urine-to-blood PCO(2) gradient to only 20 mm Hg. These subnormal PCO(2) gradient values point to proton-pump dysfunction in the collecting tubule. Histological evidence of tubulointerstitial disease accompanied the tubular defects. The striking histological feature was the presence of immunoglobulin E (IgE) deposits in glomeruli, tubuli, and vessels. Concurrent with these findings, she had high serum IgE titers and CD23 levels. IgE antibodies from her serum were reactive against human renal tubuli, with binding to two regions that matched two different proteins present in cortex and medulla. One of these proteins corresponded to carbonic anhydrase II (31 kd); the second, to an unidentified protein that seems attached to cell membranes. We suggest that these IgE antibodies could have had a pathogenic role in this patient's glomerular, tubular, and small-vessel disease. PMID- 10793032 TI - Takayasu's arteritis associated with intrarenal vessel involvement. AB - Takayasu's arteritis (TA) is an inflammatory process frequently associated with stenosis and obliteration of the aorta and its primary branches. Renal manifestations of the disease include renal artery stenosis, hypertension, and ischemic kidney disease. To our knowledge, vasculitis involving intrarenal arteries has not been reported in association with TA. We report a case with clinical and arteriographic features of TA associated with involvement of intrarenal vessels. PMID- 10793030 TI - Are selective COX-2 inhibitors nephrotoxic? AB - Nonsteroidal anti-inflammatory drugs are well-known culprits in the development of acute renal insufficiency in high-risk patients. The recent release of the selective cyclooxygenase-2 enzyme inhibitors for the treatment of inflammatory diseases and pain syndromes has been associated with a clear-cut decrease in adverse gastrointestinal effects. However, the nephrotoxic potential of these agents in patients with prostaglandin-dependent states and chronic renal impairment is unknown. Many clinicians commonly wonder if these agents can be safely prescribed to such high-risk patients. We present two cases of acute renal failure complicating the course of therapy with celecoxib in patients with chronic renal insufficiency. PMID- 10793033 TI - Sequential development of pulmonary hemorrhage with MPO-ANCA complicating anti glomerular basement membrane antibody-mediated glomerulonephritis. AB - We report a case of rapidly progressive glomerulonephritis caused by anti glomerular basement membrane (anti-GBM) antibodies that progressed to end-stage renal disease in a 67-year-old woman with diabetes. Intensive combined immunosuppressive therapy with methylprednisolone bolus, oral prednisone, and cyclophosphamide led to negativity of anti-GBM antibodies but was not able to restore renal function. After 28 months of hemodialysis, the patient suddenly presented with pulmonary hemorrhage. In this setting, high levels of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA) and negative anti-GBM antibodies were found. Therapy with oral prednisone and cyclophosphamide led to resolution of pulmonary hemorrhage and negativity of MPO-ANCA. PMID- 10793035 TI - Severe lead-induced peripheral neuropathy in a dialysis patient. AB - Toxic neuropathy caused by lead (manifested as wrist drop) was a frequent phenomenon before 1925. In modern times, it is a distinct rarity. We report herein a Hispanic woman who developed end-stage renal failure, followed by wrist drop, in whom the maximal total blood lead was 69 microg/dL. Measurements of lead in her tibia and calcaneus by K-x-ray fluorescence, however, showed markedly elevated values. The wrist drop cleared after four treatments with intravenous calcium sodium edetate (Ca EDTA). In vitro studies of (210)Pb uptake by red blood cells (RBC) after incubation with normal or uremic plasma indicated that (210)Pb uptake was inhibited by uremic plasma. These studies suggest the presence of a transport inhibitor in uremia that modifies the distribution of lead between plasma and RBC, leading to lower overall blood values. PMID- 10793034 TI - Childhood diethylene glycol poisoning treated with alcohol dehydrogenase inhibitor fomepizole and hemodialysis. AB - Diethylene glycol (DEG), a commonly used solvent, has been implicated in multiple poisoning deaths, the most recent being the Haitian acetaminophen tragedy. Unlike the more commonly seen ethylene glycol ingestion, little is understood of DEG metabolism or kinetics in humans. This has made the clinical presentation, biochemical correlates, and treatment options unclear. Patients presenting less than 12 hours after DEG ingestion may not show metabolic acidosis, whereas those presenting later may show florid metabolic acidosis. Kinetic data lend support to these observations. We report a case of DEG ingestion in a 17-month-old girl who was managed with activated charcoal, fomepizole (a recently available alcohol dehydrogenase inhibitor), and hemodialysis (HD). Pre-HD and post-HD DEG levels support clearance of DEG with HD. PMID- 10793036 TI - Symptomatic hypercalcemia of immobilization in a patient with end-stage renal disease. AB - The phenomenon of hypercalcemia in immobilization is well known, but there is limited awareness of the potential for this complication in patients with end stage renal disease (ESRD) on maintenance hemodialysis with reduced capacity for disposition of calcium. We describe such a patient who showed a calcemic response to just 3 days of immobilization in the setting of an acute illness marked by coma. Despite intensive initial therapy for hypercalcemia, including withdrawal of all calcium products and daily hemodialysis treatments using low calcium baths, her serum calcium rose to 14.0 mg/dL during the hospitalization; this metabolic abnormality appeared to perpetuate her stuporous state. Mobilization as an outpatient was the most effective therapy. Extensive testing was performed to rule out other causes for this patient's hypercalcemia. Greater recognition of acute hypercalcemia in patients with ESRD immobilized by various illnesses would preclude unnecessarily expensive and invasive testing for other causes of hypercalcemia. PMID- 10793037 TI - Prediction of hemodialysis synthetic graft thrombosis: can we identify factors that impair validity of the dysfunction hypothesis? PMID- 10793039 TI - The New Nephrologist. PMID- 10793038 TI - Are COX-2 selective inhibitors nephrotoxic? PMID- 10793041 TI - Major peptide autoepitopes for nucleosome-specific T cells of human lupus. PMID- 10793040 TI - Impact of intermittent hemodialysis variables on clinical outcomes in acute renal failure. AB - When hospital-acquired acute renal failure (ARF) is severe enough to require renal replacement therapy, mortality rates are extremely high, exceeding 50%. The potential impact of renal replacement therapy on clinical outcomes in ARF remains a subject of ongoing investigation and controversy. This article reviews in depth all of the clinical trials that have examined the effect of dialysis-related variables on clinical outcomes in patients with ARF requiring intermittent hemodialysis. In particular, the role of biocompatibility of dialyzer membranes, and timing, intensity, and adequacy of dialysis are discussed. PMID- 10793042 TI - Adult man with primary allograft nonfunction. PMID- 10793043 TI - Kidney vending: "Yes!" or "No!". PMID- 10793044 TI - Venous morphology in hemodialysis patients. PMID- 10793046 TI - Acute Rejection, Type II (Vascular). PMID- 10793045 TI - Medical treatment of cystinuria with vitamin C. PMID- 10793047 TI - Continuing Medical Education Exercise, May 2000. PMID- 10793048 TI - Strategies for improving hemodynamic stability in cardiac-compromised dialysis patients. AB - In hemodialysis patients, structural changes at all levels of the cardiovascular system are common. The presence of these cardiovascular changes is a risk factor for the development of intradialytic hypotension. This explains the clinical observation that the incidence of symptomatic hypotension is high in elderly hemodialysis patients, who often have a history of long-standing hypertension and atherosclerosis, and in hemodialysis patients with cardiovascular disease. With an increasing number of cardiovascular compromised dialysis patients, special attention should be given to this group of patients. As the age of patients on hemodialysis increases steadily, it is a challenge to provide comfortable treatment in these patients by reducing the incidence of symptomatic hypotensive periods. This article describes the use of relatively new and simple clinical maneuvers to reduce the incidence of symptomatic hypotension. PMID- 10793049 TI - Nicardipine is a safe and effective agent in pediatric hypertensive emergencies. AB - Nicardipine is the first dihydropyridine calcium channel blocker capable of intravenous administration. Seven pediatric patients with hypertensive emergencies attributable to various pathological processes were treated with intravenous nicardipine, starting at 1 microg/kg/min. Nicardipine appeared to be safe and effective in controlling hypertension in these patients. Two patients who received nicardipine through peripheral lines developed superficial thrombophlebitis. None of the five patients receiving nicardipine through a central line experienced phlebitis, and no other adverse effects were noted. PMID- 10793050 TI - Successful surgical thrombectomy of renal allograft vein thrombosis in the early postoperative period. AB - We report the successful surgical treatment of two cases of renal vein thrombosis, each occurring early after kidney transplantation. Prompt intervention was a result of accurate diagnosis by color Doppler ultrasonography. Invasive radiological procedures were not useful. Although recent advances in thrombolytic therapy have created several alternatives to open surgery, the rationale for surgical exploration in these cases was to remove the thrombus quickly, to avoid the postoperative bleeding complications of thrombolytic therapy, and to correct any technical or anatomic problems. PMID- 10793051 TI - Spontaneous improvement in a case of C1q nephropathy. AB - A 17-year-old girl showed mild proteinuria accompanied by hematuria and mild hypocomplementemia. A light microscopic study of the first renal biopsy specimen showed diffuse mild to moderate mesangial proliferation and thickening of the glomerular basement membrane (GBM). An immunofluorescence study showed dominant positive staining (3+) of IgG and C1q in the glomerular mesangium and capillary loop. Staining for C3 and fibrinogen was weak or 1+. Staining for IgA and IgM was negative. Electron-dense deposits were present in the mesangial area and also in the subepithelial, subendothelial, and intramembranous space. Urinary findings improved after dipyridamole treatment. The second renal biopsy, which was performed 5 years later, showed histological improvements, and various pictures of washing-out of deposits were also noted in an electron microscopic study. However, dominant positive staining for IgG and C1q was persistent in an immunofluorescence study. The glomerulopathy of this case belongs in the criteria of neither membranoproliferative glomerulonephritis nor lupus nephritis but could be designated as C1q nephropathy. This is the first report of a histological improvement in C1q nephropathy. PMID- 10793052 TI - A familial case of P-ANCA glomerulonephritis presenting in a father and daughter. AB - Antineutrophil cytoplasmic antibodies (ANCA) have proved to be useful serological markers for a subset of vasculitic diseases, including Wegener's granulomatosis, microscopic polyangiitis, and the Churg-Strauss syndrome. The pathogenesis of the ANCA vasculitides remains less clear, including what role, if any, genetic factors play in the expression of ANCA-associated diseases. Familial cases of systemic vasculitis have been reported, and a number of studies have addressed HLA associations of Wegener's and microscopic polyangiitis, but the results have been confusing and inconsistent. We report the first case of P-ANCA-positive vasculitis presenting in a Native American father and daughter. Both patients had systemic vasculitis and were P-ANCA positive with anti-myeloperoxidase (MPO) antibodies. PMID- 10793053 TI - Renal mucormycosis in the HIV patient. AB - Mucormycosis is an increasingly recognized opportunistic infection. It usually affects patients with debilitating conditions such as cancer, diabetes mellitus, renal failure, and extensive burns. Mucor infection has also been described in human immunodeficiency virus (HIV) patients. The most common clinical presentations are the cerebral, cutaneous, and renal forms. We describe a unique case of bilateral renal mucormycosis presenting with renal failure in an HIV infected patient. In the immunosuppressed host, a history of intravenous (IV) drug abuse associated with symptoms of pyelonephritis should alert the clinician to the possibility of mucor infection. Blood and urine culture are often negative. The diagnosis is made histologically in most cases. The treatment of HIV patients with mucormycosis and renal failure includes hemodialysis, nephrectomy, and intravenous amphotericin in addition to antiretroviral therapy. Bilateral renal involvement with Mucor carries a poor prognosis. PMID- 10793054 TI - Heavy chain deposition disease: recurrence in a renal transplant and report of IgG(2) subtype. AB - Heavy chain deposition disease (HCDD) is a rare entity characterized by tissue deposition of monoclonal heavy chains without light chains. Previous reports of HCDD include gamma(1)-, gamma(3)-, gamma(4)-, and alpha-heavy chain subtypes. Renal transplantation for HCDD has not been previously reported. We report a case of gamma(2)-HCDD in a 67-year-old patient who presented with proteinuria, hematuria, and renal insufficiency and progressed to end-stage renal failure after 6 months. The second case involves a 26-year-old woman who had a renal transplant for HCDD and recurrent gamma(1)-HCDD in the transplant. Neither patient had myeloma. The complete spectrum of gamma-HCDD subtypes has now been reported. Further data are required to make conclusive statements about the true recurrence rate of HCDD in renal transplants. PMID- 10793055 TI - Unilateral breast enlargement secondary to right brachiocephalic vein occlusion. AB - We describe a 56-year-old woman who received dialysis through a right jugular catheter and developed a progressive right breast enlargement 1 year after arteriovenous graft shunt construction in the right forearm. Arm edema was not observed. A fistulography showed retrograde long thoracic and lateral thoracic veins flow secondary to a right brachiocephalic vein occlusion. Breast enlargement disappeared completely 2 weeks after a transfemoral balloon angioplasty and stent placement. PMID- 10793057 TI - Angiotensin Receptors: What Is New? PMID- 10793056 TI - Left atrial calcification in a hemodialysis patient with cor triatriatum. AB - Myocardial calcification is a rare manifestation of abnormal calcium metabolism seen in some patients with chronic renal failure. This report describes the transesophageal echocardiographic and spiral computed tomography (CT) findings in a young hemodialysis female with severe secondary hyperparathyroidism. These findings included calcification of the multiperforated membrane of a cor triatriatum and the wall of the left atrium. PMID- 10793058 TI - Regulation of CD30 antigen expression and its potential significance for human disease. PMID- 10793059 TI - Heme oxygenase-1 in tissue pathology: the Yin and Yang. PMID- 10793061 TI - Vascular endothelial growth factor receptor-3 in lymphangiogenesis in wound healing. AB - Vascular endothelial growth factor receptor-3 (VEGFR-3) is essential for embryonic cardiovascular development, but thereafter becomes confined to the lymphatic endothelium in adult tissues. We have here studied VEGFR-3 expression in experimental wounds of pigs and chronic inflammatory wounds of humans. In healing incisional and punch biopsy wounds made in the dorsal skin of pigs, angiogenic blood vessels, identified by use of the blood vascular endothelial markers vWF and PAL-E and the basal lamina protein laminin, developed into the granulation tissue stroma from day 4 onward, being most abundant on days 5 and 6 and regressing thereafter. VEGFR-3-positive vessels were observed in the granulation tissue from day 5 onward. These vessels were distinct from the PAL E/laminin/vWF-positive vessels and fewer in number, and they appeared to sprout from pre-existing VEGFR-3-positive lymphatic vessels at the wound edge. Unlike the blood vessels, very few VEGFR-3-positive lymphatic vessels persisted on day 9 and none on day 14. In chronic wounds such as ulcers and decubitus wounds of the lower extremity of humans, VEGFR-3 was also weakly expressed in the vascular endothelium. Our results suggest that transient lymphangiogenesis occurs in parallel with angiogenesis in healing wounds and that VEGFR-3 becomes up regulated in blood vessel endothelium in chronic inflammatory wounds. PMID- 10793062 TI - Immunohistochemical analysis of cyclin D1 shows deregulated expression in multiple myeloma with the t(11;14). AB - The t(11;14)(q13;q32) chromosomal translocation, the hallmark of mantle cell lymphoma (MCL), is recurrently found in multiple myelomas (MM) by means of conventional cytogenetics. Unlike MCL, recent molecular studies of MM-derived cell lines with t(11;14) have indicated that the breakpoints are highly dispersed over the 11q13 region; however, the fact that cyclin D1 is generally overexpressed in these cell lines suggests that this gene is the target of the translocation. To evaluate further the involvement of cyclin D1 in MM, we used immunohistochemistry and fluorescence in situ hybridization to investigate cyclin D1 expression and the presence of chromosome 11 abnormalities in a representative panel of 48 MM patients (40 at diagnosis and 8 at relapse). Cyclin D1 overexpression occurred in 12/48 (25%) of cases; combined immunohistochemistry and fluorescence in situ hybridization analyses in 39 patients showed cyclin D1 positivity in all of the cases (7/7) bearing the t(11;14), in two of the 13 cases with trisomy 11, and in one of the 19 cases with no apparent abnormalities of chromosome 11. Our data indicate that the t(11;14) translocation in MM leads to cyclin D1 overexpression and that immunohistochemical analysis may represent a reliable means of identifying this lesion in MM. PMID- 10793060 TI - Regulation of tissue injury responses by the exposure of matricryptic sites within extracellular matrix molecules. AB - Extracellular matrix (ECM) is known to provide signals controlling cell shape, migration, proliferation, differentiation, morphogenesis, and survival. Recent data shows that some of these signals are derived from biologically active cryptic sites within matrix molecules (matricryptic sites) that are revealed after structural or conformational alteration of these molecules. We propose the name, matricryptins, for enzymatic fragments of ECM containing exposed matricryptic sites. Mechanisms regulating the exposure of matricryptic sites within ECM molecules include the major mechanism of enzymatic breakdown as well as others including ECM protein multimerization, adsorption to other molecules, cell-mediated mechanical forces, and ECM denaturation. Such matrix alterations occur during or as a result of tissue injury, and thus, the appearance of matricryptic sites within an injury site may provide important new signals to regulate the repair process. Here, we review the data supporting this concept and provide insight into why the increased exposure of matricryptic sites may be an important regulatory step in tissue responses to injury. PMID- 10793063 TI - E-cadherin expression in melanoma cells restores keratinocyte-mediated growth control and down-regulates expression of invasion-related adhesion receptors. AB - In human epidermis, functional symbiosis requires homeostatic balance between keratinocytes and melanocytes. Compelling evidence from co-culture studies demonstrated a sophisticated, multileveled regulation of normal melanocytic phenotype orchestrated by undifferentiated, basal-type keratinocytes. Keratinocytes control cell growth and dendricity, as well as expression of melanoma-associated cell surface molecules of normal melanocytes. In contrast, melanoma cells are refractory to the keratinocyte-mediated regulation. The loss of regulatory dominance by keratinocytes occurs in concert with down-regulation of E-cadherin expression in melanoma cells. To investigate the potential role of E-cadherin in melanoma-keratinocyte interaction, we transduced E-cadherin negative melanoma cells with full-length E-cadherin cDNA using an adenoviral vector. Our results show that functional E-cadherin expression in melanoma cells leads to cell adhesion to keratinocytes rendering them susceptible for keratinocyte-mediated control. In a skin reconstruction model, ectopic E-cadherin expression inhibits invasion of melanoma cells into dermis by down-regulating invasion-related adhesion receptors, MelCAM/MUC18 and beta3 integrin subunit, and by induction of apoptosis. Thus, disruption of the E-cadherin-mediated, normal regulatory control from keratinocytes may represent one of the mechanisms accounting for melanocyte transformation. PMID- 10793064 TI - The indispensability of heme oxygenase-1 in protecting against acute heme protein induced toxicity in vivo. AB - Heme oxygenase (HO) is the rate limiting enzyme in the degradation of heme, and its isozyme, HO-1, may protect against tissue injury. One posited mechanism is the degradation of heme released from destabilized heme proteins. We demonstrate that HO-1 is a critical protectant against acute heme protein-induced toxicity in vivo. In the glycerol model of heme protein toxicity-one characterized by myolysis, hemolysis, and kidney damage-HO-1 is rapidly induced in the kidney of HO-1 +/+ mice as the latter sustain mild, reversible renal insufficiency without mortality. In stark contrast, after this insult, HO-1 -/- mice exhibit fulminant, irreversible renal failure and 100% mortality; HO-1 -/- mice do not express HO-1, and evince an eightfold increment in kidney heme content as compared to HO-1 +/+ mice. We also demonstrate directly the critical dependency on HO-1 in protecting against a specific heme protein, namely, hemoglobin: doses of hemoglobin which exert no nephrotoxicity or mortality in HO-1 +/+ mice, however, precipitate rapidly developing, acute renal failure and marked mortality in HO-1 -/- mice. We conclude that the induction of HO-1 is an indispensable response in protecting against acute heme protein toxicity in vivo. PMID- 10793066 TI - Complement activation after oxidative stress: role of the lectin complement pathway. AB - The complement system plays an important role in mediating tissue injury after oxidative stress. The role of mannose-binding lectin (MBL) and the lectin complement pathway (LCP) in mediating complement activation after endothelial oxidative stress was investigated. iC3b deposition on hypoxic (24 hours; 1% O(2))/reoxygenated (3 hours; 21% O(2)) human endothelial cells was attenuated by N-acetyl-D-glucosamine or D-mannose, but not L-mannose, in a dose-dependent manner. Endothelial iC3b deposition after oxidative stress was also attenuated in MBL-deficient serum. Novel, functionally inhibitory, anti-human MBL monoclonal antibodies attenuated MBL-dependent C3 deposition on mannan-coated plates in a dose-dependent manner. Treatment of human serum with anti-MBL monoclonal antibodies inhibited MBL and C3 deposition after endothelial oxidative stress. Consistent with our in vitro findings, C3 and MBL immunostaining throughout the ischemic area at risk increased during rat myocardial reperfusion in vivo. These data suggest that the LCP mediates complement activation after tissue oxidative stress. Inhibition of MBL may represent a novel therapeutic strategy for ischemia/reperfusion injury and other complement-mediated disease states. PMID- 10793065 TI - Genetic and epigenetic changes in human epithelial cells immortalized by telomerase. AB - Exogenous expression of hTERT, the catalytic component of telomerase, is sufficient for the immortalization of human fibroblasts but insufficient for the immortalization of human foreskin keratinocytes (HFKs) and human mammary epithelial cells (HMECs). These latter cell types can overcome senescence by coexpression of hTERT and human papillomavirus (HPV) E7 or by expression of hTERT and loss of p16(INK4a) expression, indicating that the retinoblastoma (Rb) pathway, along with a telomere maintenance pathway, plays a role in determining the life span of epithelial cells. In this study, we further characterize hTERT immortalized HFKs and human adenoid epithelial cells (HAKs) for genotypic and phenotypic alterations that are associated with immortalization. Of five hTERT immortalized HFK and HAK cell lines examined, four exhibited repression of p16(INK4a) expression by promoter methylation or specific large-scale deletion of chromosome 9p, the location of p16(INK4a). Interestingly, one cell line exhibited complete down-regulation of expression of p14(ARF), with only slight down regulation of expression of p16(INK4a). Yet, all of the immortal cells lines exhibited hyperphosphorylated Rb. Cytogenetic analysis revealed clonal chromosome aberrations in three of the five cell lines. All of the cell lines retained a growth block response with the expression of mutant ras. When grown on organotypic raft cultures, however, the hTERT-immortalized cells exhibited a maturation delay on terminal differentiation. Our results indicate that immortalization of epithelial cells may require both activation of telomerase and other genetic and/or epigenetic alterations that abrogate normal differentiation. PMID- 10793067 TI - Severe destructive autoimmune lesions with aging in murine Sjogren's syndrome through Fas-mediated apoptosis. AB - When we evaluated the age-associated changes in autoimmune exocrinopathy in a NFS/sld murine model for primary Sjogren's syndrome (SS), severe destructive autoimmune lesions developed in the salivary and lacrimal glands in the aged mice, compared with those observed in the younger model. We detected a decreased secretion of saliva and tear flow in the aged group. A significant increase of TUNEL(+)-apoptotic epithelial duct cells in the salivary glands was detected in the aged SS animal model. A higher proportion of mouse salivary gland cells bearing Fas was found in the aged group, whereas no significant changes were seen on tissue-infiltrating CD4(+) T cells bearing FasL in the salivary glands from young and aged mice. We detected an increased cleavage product of organ-specific autoantigen, 120-kd alpha-fodrin, in the aged salivary gland tissues on immunoblotting, and an increase in serum autoantibody production against 120-kd alpha-fodrin by enzyme-linked immunosorbent assay. An increase in the proliferative response of splenic T cells against organ-specific autoantigen was observed, whereas nonspecific concanavalin A responsiveness was decreased in the aged mice. In addition, a decrease in Fas expression was found on splenic CD4(+) T cells in the aged mice, and anti-Fas mAb-stimulated apoptosis was down regulated on CD4(+) T cells. These results indicate that age-associated dysregulation of CD4(+) T cells may play a crucial role on acceleration of organ specific autoimmune lesions in a murine model for primary SS through Fas-mediated apoptosis. PMID- 10793068 TI - p16(INK4a) gene alterations are frequent in lesions of mycosis fungoides. AB - Mycosis fungoides is usually an indolent disease that, after a variable period of time in a stable phase, evolves into a tumoral form with aggressive behavior. The molecular events that mark this progression remain essentially unknown to date, and this prompted us to investigate the hypothetical role of p16(INK4a) silencing in mycosis fungoides progression. We analyzed the three most frequent mechanisms of inactivation of the p16(INK4a) gene (deletion, promoter hypermethylation, and mutation) in nine cases with patch/plaque and tumoral lesions of mycosis fungoides. The existence of alterations was investigated by microsatellite analysis, methylation-specific polymerase chain reaction, and direct sequencing. Alterations of the p16(INK4a) gene were found in four of nine of the plaque lesions (hypermethylation in three samples and allelic loss in one sample) and seven of nine in the tumor lesions (hypermethylation in five samples and allelic loss in three samples). No case presented point mutations. Although a higher incidence of p16(INK4a) hypermethylation was observed in the cases that failed to achieve a complete remission, the limited size of our series prompted us to evaluate this finding cautiously. The results of this study therefore showed a common genetic alteration that is found more frequently in tumoral lesions than it is in plaque lesions of mycosis fungoides. It also offers data that could suggest a relationship between p16(INK4a) hypermethylation and unfavorable clinical outcome. Broader studies are needed to confirm both relationships. PMID- 10793070 TI - Degranulating eosinophils in human endometriosis. AB - Degranulating eosinophils have been described in most endometrial cancers. We hypothesized that endometriosis (ectopic, nonneoplastic endometrial tissue) would be an appropriate model system for determining whether eosinophil degranulation is part of a specific immune response to endometrial cancer or if it is related to the more general phenomenon of tissue remodeling (wound healing) that is common to both disorders. To test this hypothesis, we performed immunohistochemistry and Western blotting to evaluate the presence of eosinophil peroxidase (a marker of eosinophil degranulation) in normal endometrium (n = 20) and endometriosis samples (n = 24) and to define the coexpression of three eosinophil chemoattractants: interleukin-5 (IL-5), eotaxin, and regulated on activator-normal T cell expressed and secreted (RANTES). There was focally intense deposition of eosinophil peroxidase in the fibrotic connective tissue and blood vessels of 21 of 24 human endometriosis specimens; two samples showed weak staining, and only one tissue was negative for eosinophil degranulation. None of the 10 normal proliferative endometrial specimens had evidence of eosinophil degranulation, and four of 10 secretory tissues stained only weakly for eosinophil peroxidase. The presence of degranulating eosinophils was also associated with the presence of eotaxin and IL-5 in some samples and with RANTES in others. We conclude that the abundant presence of degranulating eosinophils in the fibrous regions of endometriosis supports the interpretation that eosinophils are involved in general tissue remodeling and wound healing rather than a tissue directed immune response. PMID- 10793069 TI - Expression of p16/INK4a in posttransplantation lymphoproliferative disorders. AB - It was recently demonstrated that classification of posttransplantation lymphoproliferative disorders (PT-LPDs) into morphological and molecular categories is clinically relevant. It was also reported that PT-LPD not associated with Epstein-Barr virus (EBV) had a more aggressive course than most lesions associated with EBV. Because the cyclin-dependent kinase inhibitor p16/INK4a has been reported to be frequently inactivated in high-grade lymphomas, we evaluated 17 PT-LPD to determine whether p16/INK4a expression could be correlated to morphology, EBV detection, and a Ki-67 labeling index. We demonstrated that tumors with no p16/INK4a expression (n = 8) had a predominantly monomorphic appearance, and most were EBV negative (respectively, 7/8 and 5/8), whereas lesions with p16/INK4a expression (n = 9) were mostly polymorphic PT-LPD (6/9) (P = 0.049) and associated with EBV (9/9) (P = 0.015). In particular, strong p16/INK4a expression was observed in atypical immunoblasts and Reed Sternberg-like cells. Furthermore, the proliferation index was significantly higher in tumors lacking p16/INK4a expression than in other lesions (P = 0.0008). In conclusion, down-regulation of p16/INK4a was mostly observed in PT-LPD lesions known to follow more aggressive courses: monomorphic tumors and EBV-negative PT neoplasms. Conversely, overexpression of p16/INK4a was associated with EBV positive PT-LPD. While p16/INK4a might play a role in the proliferative rate of LP-LPD, further investigations are needed to assess the clinical relevance of p16/INK4a expression in predicting the evolution of tumors and to explain how EBV could favor p16/INK4a protein accumulation in lesions. PMID- 10793071 TI - Contribution of apoptosis and apoptosis-related proteins to the malformation of the primitive intrahepatic biliary system in Meckel syndrome. AB - In the developing liver, the complete or partial persistence of the primitive double-layered cylinder of biliary-type cells that surrounds the branches of portal vein and its mesenchyme gives origin to portal tracts with an increased number of bile duct structures. The term "ductal plate malformation of the liver" was coined to label the insufficient remodeling of the primitive intrahepatic biliary system. Meckel syndrome is an autosomal recessive inherited disease characterized by occipital encephalocele, postaxial polydactyly, diffuse cystic renal dysplasia, and malformation of the ductal plate of the liver. We studied 52 fetuses with Meckel syndrome from five German centers (Berlin, Freiburg, Heidelberg, Mainz, and Marburg). Analysis of apoptosis and cell proliferation (Ki 67) was performed by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) and immunohistochemistry in the liver of 24 normal fetuses of different gestational ages (14-38 weeks of gestation) and in 14 fetuses with Meckel syndrome (17-38 weeks of gestation). The expression of two apoptosis related proteins, Fas (a transmembrane cell surface protein involved in the apoptosis) and Bcl-2 (an anti-apoptotic protein), was studied by immunohistochemistry in the liver of 11 normal fetuses of different gestational ages (14-40 weeks of gestation) and in 40 fetuses with Meckel syndrome (16-38 weeks of gestation). In control fetuses, apoptosis rate and cell proliferation were high in the remodeling ductal plate and moderate in the ductal plate and in remodeled bile ducts. During gestation, expression of Fas and Bcl-2 decreased and increased, respectively. The malformed ductal plates in the fetal livers with Meckel syndrome showed a marked decrease in the apoptotic rate and Fas expression and an increase in proliferative activity and Bcl-2 expression in comparison with control fetuses. Furthermore, by linear regression analysis, we found that both proliferation activity and apoptosis rate in the ductal plate malformation of fetuses with Meckel syndrome were practically constant along the gestation. These results, which represent the first systematic study of apoptosis in ductal plate malformation of the liver, indicate that 1) animals harboring the gene defect of Meckel syndrome could be a good model for the study of the abnormal development of the primitive intrahepatic biliary system, 2) a decreased cell turnover occurs in the ductal plate malformation of fetuses with Meckel syndrome, and 3) the increase of Bcl-2 expression contributes to the pathogenesis of the lack of remodeling of ductal plate of the liver in Meckel syndrome. PMID- 10793072 TI - Characterization and isolation of ductular cells coexpressing neural cell adhesion molecule and Bcl-2 from primary cholangiopathies and ductal plate malformations. AB - It has recently been shown that reactive bile ductules display neuroendocrine features, including immunoreactivity for the neural cell adhesion molecule (NCAM). In this study we have compared the immunohistochemical expression of NCAM with that of HEA-125 (biliary specific) and LKM-1 (hepatocyte specific) and other markers relevant to morphogenesis (Bcl-2, EMA) and cell proliferation (Ki-67) in cryostat sections from different chronic liver diseases and from fetal livers at different gestational ages. In parallel, viable NCAM-positive ductular cells were purified from collagenase digests of cirrhotic livers by immunomagnetic separation and characterized by immunocytochemistry and transmission electron microscopy. We demonstrated that reactive ductules with atypical morphology coexpressed NCAM and Bcl-2 and were found mainly in congenital diseases associated with ductal plate malformation and in primary cholangiopathies. On the contrary, reactive ductules with typical morphology were negative for NCAM/Bcl-2 and positive for EMA. Reactive ductules coexpressing NCAM/Bcl-2 were negative for the proliferation marker Ki-67 and appeared to be directly connected with periportal hepatocytes. In fetal livers NCAM/Bcl-2 was transiently expressed during the early developmental stages of ductal plate (10-16 weeks) and started to disappear as the ductal plate began duplicating. NCAM-positive ductal plate cells were Ki-67 negative, becoming positive in duplicated segments. Thus the histogenesis of ductular reactive cells seems to recapitulate the early stages of biliary ontogenesis. In primary cholangiopathies and ductal plate malformations, these cells do not appear to maturate further, and thus abundant ductular structures coexist with vanishing mature ducts. These NCAM-positive ductular cells were immunopurified from patients with chronic cholestatic liver diseases and showed ultrastructural features consistent with a less differentiated phenotype than mature cholangiocytes. These isolated cells represent a useful model for in vitro studies. PMID- 10793073 TI - Hepatocellular alterations after intraportal transplantation of ovarian tissue in ovariectomized rats. AB - The mechanisms of hepatocarcinogenesis by certain synthetic estrogens seem to involve both nongenotoxic and indirect genotoxic effects. However, the natural estrogen estradiol did not exert any carcinogenic effects in established experimental protocols. To elucidate specific long-term effects of natural estrogens on hepatocytes, small pieces of ovarian tissue were transplanted via the portal vein into the livers of ovariectomized female rats. One week, 3 weeks, and 3 months after transplantation the transplants were found to proliferate and to secrete estradiol. Three weeks after transplantation the hepatocytes of the liver acini downstream of the stimulated transplants already showed a remarkable loss of glycogen, distinct cytoplasmic amphophilia, enlargement of their nuclei, a strong increase in the number and size of peroxisomes, an increase in proliferative activity and apoptotic elimination, and changes in the activity of certain key enzymes of energy metabolism. All hepatocellular alterations could be inhibited by the estrogen receptor antagonist toremifene and are, therefore, attributed to specific effects of estradiol produced by the transplants. The observed alterations resemble in some respects amphophilic preneoplastic liver foci, which particularly occur after long-term administration of nongenotoxic hepatocarcinogens, including the adrenal steroid hormone dehydroepiandrosterone. In a preliminary experiment three of six animals exhibited a hepatocellular carcinoma, and another animal developed a hepatocellular adenoma 18 months after intrahepatic ovarian tissue transplantation. PMID- 10793076 TI - Immunocytochemical localization of metallothionein and its relation to doxorubicin toxicity in transgenic mouse heart. AB - Previous studies using a cardiac-specific metallothionein-overexpressing transgenic mouse model have demonstrated that metallothionein protects the heart from doxorubicin toxicity. The present study was undertaken to determine cellular and subcellular distribution of metallothionein and located the antioxidant action of this protein in the transgenic heart. Using light microscopic immunoperoxidase method, it was identified that the overexpressed metallothionein is localized exclusively in cardiomyocytes. The electron microscopic immunogold method revealed that elevated metallothionein is in nucleus, myofibers, and sarcoplasm. In contrast with these distributions, metallothionein in nontransgenic myocardium was undetectable by immunoperoxidase light microscopy and was seldom found in nucleus and myofibers by immunogold electron microscopy. Treatment with doxorubicin induced cytoplasmic vacuolization and severe damages in myofilaments and nucleus in nontransgenic myocardium. The most prominent injury, however, occurred in mitochondria, including striking size and shape changes, focal swelling and loss of cristae. These damages were rarely found in the doxorubicin-treated transgenic myocardium. In particular, the internal morphology of mitochondria was maintained essentially normal, although metallothionein was not localized in this compartment in transgenic hearts. This study thus demonstrates that although the subcellularly localized action of metallothionein is important, it also plays a significant role in protection against oxidative injury by doxorubicin in remote organelles. PMID- 10793074 TI - The development and compensation of biliary cirrhosis in interleukin-6-deficient mice. AB - In an effort to understand the role of IL-6/gp130 signaling in chronic liver injury, IL-6 deficient (IL-6(-/-)) and wild-type control (IL-6(+/+)) mice were subjected to bile duct ligation (BDL) for 12 weeks. This maneuver causes chronic biomechanical stress and liver injury, fueling sustained biliary epithelial and hepatocyte proliferation. By 12 weeks after BDL, IL-6(-/-) mice develop significantly higher total serum bilirubin levels (23.2 +/- 2.3 versus 14.9 +/- 2.1 mg/dl, P < 0.0001; delta bilirubin subfraction 16.7 +/- 4.0% versus 9.2 +/- 1.8%; P < 0.002), and the majority (15/18) show "black" gallbladder bile, compared to IL-6(+/+) mice (5/16; P < 0.003). The IL-6(-/-) mice also cannot sustain the compensatory liver mass increase commonly seen with chronic obstructive cholangiopathy, because of less hepatocyte proliferation, despite a rate of hepatocyte apoptosis similar to that of IL-6(+/+) mice. Moreover, IL-6(-/ ) mice show a more advanced stage of biliary fibrosis and a higher mortality rate than the IL-6(+/+) controls (51% versus 23%; P < 0.02). These phenotypic changes in the IL-6(-/-) mice are associated with decreased expression and phosphorylation of gp130 and the transcription factor STAT3, compared to IL 6(+/+) mice. Daily treatment with exogenous recombinant IL-6 for 3-6 weeks starting at 6 weeks after BDL significantly lowers the serum total bilirubin in both groups. In the IL-6(-/-) mice, exogenous IL-6 treatment also increases the level of gp130 protein expression and completely reverses the loss of liver mass by increasing the hepatocyte proliferation. In conclusion, IL-6 appears to contribute to biliary tree integrity and maintenance of hepatocyte mass during chronic injury. PMID- 10793075 TI - Genetic, immunohistochemical, and clinical features of medullary carcinoma of the pancreas: A newly described and characterized entity. AB - Medullary carcinomas of the pancreas are a recently described, histologically distinct subset of poorly differentiated adenocarcinomas that may have a unique pathogenesis and clinical course. To further evaluate these neoplasms, we studied genetic, pathological, and clinical features of 13 newly identified medullary carcinomas of the pancreas. Nine (69%) of these had wild-type K-ras genes, and one had microsatellite instability (MSI). This MSI medullary carcinoma, along with three previously reported MSI medullary carcinomas, were examined immunohistochemically for Mlh1 and Msh2 expression, and all four expressed Msh2 but did not express Mlh1. In contrast, all of the medullary carcinomas without MSI expressed both Msh2 and Mlh1. Remarkably, the MSI medullary carcinoma of the pancreas in the present series arose in a patient with a synchronous but histologically distinct cecal carcinoma that also had MSI and did not express Mlh1. The synchronous occurrence of two MSI carcinomas suggests an inherited basis for the development of these carcinomas. Indeed, the medullary phenotype, irrespective of MSI, was highly associated with a family history of cancer in first-degree relatives (P < 0.001). Finally, one medullary carcinoma with lymphoepithelioma-like features contained Epstein-Barr virus-encoded RNA-1 by in situ hybridization. Therefore, because of medullary carcinoma's special genetic, immunohistochemical, and clinical features, recognition of the medullary variant of pancreatic adenocarcinoma is important. Only by classifying medullary carcinoma as special subset of adenocarcinoma can we hope to further elucidate its unique pathogenesis. PMID- 10793077 TI - Up-regulation of AT(1) and AT(2) receptors in postinfarcted hypertrophied myocytes and stretch-mediated apoptotic cell death. AB - To determine whether up-regulation of AT(1) and AT(2) receptors occurred in hypertrophied myocytes after infarction and whether AT(2) played a role in stretch-mediated apoptosis, left ventricular myocytes were dissociated from the surviving portion of the wall 8 days after coronary occlusion and cardiac failure in rats. Control cells were obtained from sham-operated animals. Myocytes were stretched in an equibiaxial stretch apparatus and angiotensin II (Ang II) formation and cell death were measured 3 and 12 hours later. AT(1) and AT(2) proteins were evaluated in freshly isolated myocytes and after stretch. The effects of AT(1) and AT(2) antagonists on stretch-induced Ang II synthesis and apoptosis were also established. Myocardial infarction increased AT(1) and AT(2) in myocytes and stretch further up-regulated these receptors. Ang II levels were higher in postinfarcted myocytes and this peptide increased with the duration of stretch in both groups of cells. Similarly, apoptosis increased with time in control and postinfarcted myocytes. Absolute values of Ang II and apoptosis were greater in myocytes from infarcted hearts at 3 and 12 hours after stretch. Addition of AT(1) blocker to cultures inhibited stretch-activated apoptosis in both myocyte populations as well as the generation of Ang II in postinfarcted myocytes. In contrast, AT(2) antagonists had no impact on these cellular events. In conclusion, Ang II stimulated cell death through AT(1) receptor activation, whereas ligand binding to AT(2) receptor did not alter Ang II concentration and apoptosis in normal and postinfarcted hypertrophied myocytes. PMID- 10793078 TI - RGD-dependent vacuolation and lumen formation observed during endothelial cell morphogenesis in three-dimensional fibrin matrices involves the alpha(v)beta(3) and alpha(5)beta(1) integrins. AB - Recent data have revealed the involvement of the alpha(v)beta(3) integrin in angiogenesis. However, few studies to date have provided a convincing role for this receptor in in vitro assays of endothelial cell morphogenesis where defined steps can be examined. Here, we present data showing that two integrins, alpha(v)beta(3) and alpha(5)beta(1), regulate human endothelial cell vacuolation and lumen formation in three-dimensional fibrin matrices. Cells resuspended in fibrin formed intracellular vacuoles that coalesced into lumenal structures. These morphogenic events were completely inhibited by the simultaneous addition of anti-alpha(v)beta(3) and anti-alpha(5) integrin antibodies. Complete blockade was also accomplished with a combination of the cyclic Arg-Gly-Asp (cRGD) peptide and anti-alpha(5) integrin antibodies. No blockade was observed with the control Arg-Gly-Glu (RGE) peptide alone or in combination with control antibodies. Finally, we were able to demonstrate regression of vacuoles and lumens several hours after the addition of cRGD peptides combined with anti-alpha(5) integrin antibodies. These effects were not observed with control peptides alone or in combination with control antibodies. We report here the novel involvement of both the alpha(v)beta(3) and alpha(5)beta(1) integrins in vacuolation and lumen formation in a fibrin matrix, implicating a role for multiple integrins in endothelial cell morphogenesis. PMID- 10793079 TI - Decreased expression of mouse Rbm3, a cold-shock protein, in Sertoli cells of cryptorchid testis. AB - Physiological scrotal hypothermia is necessary for normal spermatogenesis and fertility in mammals. Human RNA binding motif protein 3 (RBM3) is structurally highly similar to the cold-inducible RNA-binding protein (Cirp), and both mRNAs are induced in human cells at the scrotal temperature (32 degrees C). We report here the cloning of mouse Rbm3 cDNA, which encoded an 18-kd protein with 94% identity in amino acid sequence to that of human RBM3. In the testis of adult mice, Rbm3 mRNA and protein were detected in Sertoli cells, but not germ cells, of seminiferous tubules at all stages. The expression was not observed in Sertoli cells of fetuses, but was observed in newborn and older mice. In the TAMA26 mouse Sertoli cell line, the Rbm3 expression level was increased or decreased within 12 hours after temperature shift from 37 degrees C to 32 degrees C or 39 degrees C, respectively. In contrast to Cirp, the cold-induced growth suppression of TAMA26 cells was not affected by suppression of the Rbm3 expression. When mouse testis was exposed to heat stress by experimental cryptorchidism, the level of Rbm3 was decreased in Sertoli cells. Rbm3 may play important roles distinct from those played by Cirp in spermatogenesis and cryptorchidism by regulating the gene expression in Sertoli cells. PMID- 10793081 TI - Proliferative activity of intratumoral fibroblasts is closely correlated with lymph node and distant organ metastases of invasive ductal carcinoma of the breast. AB - Mitotic figures of fibroblasts are seen within invasive ductal carcinoma (IDC) of the breast. This suggests that the proliferative activity of fibroblasts may play an important role in IDC tumor progression. The purpose of this study was to examine whether the proliferative activity of fibroblasts can predict lymph node metastasis (LNM) or distant-organ metastasis (DOM) of IDCs. Two hundred four consecutive patients with IDC of the breast surgically treated at the National Cancer Center Hospital East constituted the basis of this study. Proliferative activity of fibroblasts was immunohistochemically evaluated by the mouse MIB-1 monoclonal antibody against Ki-67 antigen. The MIB-1 labeling index was the percentage of fibroblasts with positively stained nuclei, and fields for cell counting were selected in inner and outer areas within IDCs. In both areas, 300 fibroblasts were counted in each high-power field. The significance of proliferative activity of fibroblasts on LNM or DOM was compared with well-known prognostic parameters. Multivariate analyses demonstrated that a MIB-1 labeling index of more than 10% of fibroblasts in the inner area of IDCs significantly increased the relative risk of LNM and hazard rate of DOM (P < 0.001 and P = 0.007, respectively). The present study indicated that the metastatic ability of IDCs is closely dependent on proliferative activity of fibroblasts in the inner area. PMID- 10793080 TI - Differential nuclear and cytoplasmic expression of PTEN in normal thyroid tissue, and benign and malignant epithelial thyroid tumors. AB - Germline mutations in PTEN (MMAC1/TEP1) are found in patients with Cowden syndrome, a familial cancer syndrome which is characterized by a high risk of breast and thyroid neoplasia. Although somatic intragenic PTEN mutations have rarely been found in benign and malignant sporadic thyroid tumors, loss of heterozygosity (LOH) has been reported in up to one fourth of follicular thyroid adenomas (FAs) and carcinomas. In this study, we examined PTEN expression in 139 sporadic nonmedullary thyroid tumors (55 FA, 27 follicular thyroid carcinomas, 35 papillary thyroid carcinomas, and 22 undifferentiated thyroid carcinomas) using immunohistochemistry and correlated this to the results of LOH studies. Normal follicular thyroid cells showed a strong to moderate nuclear or nuclear membrane signal although the cytoplasmic staining was less strong. In FAs the neoplastic nuclei had less intense PTEN staining, although the cytoplasmic PTEN-staining intensity did not differ significantly from that observed in normal follicular cells. In thyroid carcinomas as a group, nuclear PTEN immunostaining was mostly weak in comparison with normal thyroid follicular cells and FAs. The cytoplasmic staining was more intense than the nuclear staining in 35 to 49% of carcinomas, depending on the histological type. Among 81 informative tumors assessed for LOH, there seemed to be an associative trend between decreased nuclear and cytoplasmic staining and 10q23 LOH (P = 0.003, P = 0.008, respectively). These data support a role for PTEN in the pathogenesis of follicular thyroid tumors. PMID- 10793082 TI - Expression of the ALK tyrosine kinase gene in neuroblastoma. AB - ALK (anaplastic lymphoma kinase) is a tyrosine kinase receptor, expressed as part of the chimeric NPM-ALK protein, in anaplastic large cell lymphomas (ALCLs) exhibiting the t(2;5)(p23;q35) translocation. As a result of this translocation, the NPM (nucleophosmin) gene is fused to the portion of the ALK gene encoding its intracytoplasmic segment. In normal mouse tissues, mRNA encoding the Alk receptor has been found only in neural cells, suggesting involvement of this receptor in the development of the nervous system. The purpose of the present study was to examine the presence of ALK transcripts and protein in normal human tissues and a variety of cell lines and human tumors. Emphasis was placed on neuroblastomas because other tyrosine kinase receptors are expressed in human neuroblastomas. Fifty-six cell lines, including 29 lines of neural origin, and lymphoid and nonlymphoid tissue specimens, including 24 neuroblastomas, were investigated for ALK expression, using reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry. The results confirmed that mRNA encoding ALK protein was not detectable in any normal or neoplastic hematopoietic tissue tested, except for t(2;5)-positive ALCL. The salient finding was that 13 of the 29 cell lines of neural origin and 22 of 24 neuroblastomas were found to express ALK transcripts and ALK protein. However, no correlation was evident between any known prognostic factors and the level of ALK expression. PMID- 10793083 TI - Involvement of Sp1 and microsatellite repressor sequences in the transcriptional control of the human CD30 gene. AB - CD30, as a member of the tumor necrosis factor (TNF) receptor family, is expressed on the surface of activated lymphoid cells. CD30 overexpression is a characteristic of lymphoproliferative diseases such as Hodgkin's/non-Hodgkin's lymphomas, embryonal carcinoma, and a number of Th2-associated diseases. The CD30 gene has been mapped to a region of the murine genome that is involved in susceptibility to systemic lupus erythematosus. Functionally, CD30 may play a role in the deletion of autoreactive T cells. We were interested in determining the molecular nature of CD30 overexpression. Sequence comparison has revealed significant identity between the TATA-less human and murine CD30 promoters; they share a number of common consensus binding motifs. Transfection assays identified three regions of transcriptional importance; the region between position -1.2 kb and -336 bp, containing a CCAT microsatellite sequence, a conserved Sp1 site at positions -43 to -38, and a downstream promoter element (DPE) at positions +24 to +29. EMSA and DNase I footprinting showed specific DNA-protein interactions of the CD30 promoter with the Sp1 site and the CCAT repeat region. The DPE element was shown to be essential for start site selection. We conclude that the conserved Sp1 site at -43 to -38 is associated with maximum reporter gene activity, the DPE element is required for start site selection, and the CCAT tetranucleotide repeats act to repress transcription. We also have shown that the microsatellite is multiallelic, when we screened a random healthy population. Further studies are required to determine whether microsatellite instability in the repressor predisposes susceptible individuals to CD30 overexpression. PMID- 10793084 TI - Vascular endothelial growth factor (VEGF)-induced retinal vascular permeability is mediated by intercellular adhesion molecule-1 (ICAM-1). AB - Two prominent vascular endothelial growth factor (VEGF)-induced retinal effects are vascular permeability and capillary nonperfusion. The mechanisms by which these effects occur are not completely known. Using a rat model, we show that intravitreous injections of VEGF precipitate an extensive retinal leukocyte stasis (leukostasis) that coincides with enhanced vascular permeability and capillary nonperfusion. The leukostasis is accompanied by the up-regulation of intercellular adhesion molecule-1 expression in the retina. The inhibition of intercellular adhesion molecule-1 bioactivity with a neutralizing antibody prevents the permeability and leukostasis increases by 79% and 54%, respectively. These data are the first to demonstrate that a nonendothelial cell type contributes to VEGF-induced vascular permeability. Additionally, they identify a potential mechanism for VEGF-induced retinal capillary nonperfusion. PMID- 10793085 TI - Strain-dependent vascular remodeling phenotypes in inbred mice. AB - We have recently established a mouse model of arterial remodeling in which flow in the left common carotid artery of FVB mice was interrupted by ligation of the vessel near the carotid bifurcation, resulting in a dramatic reduction of the lumen as a consequence of a reduction in vessel diameter and intimal lesion formation. In the present study we applied this model to various inbred strains of mice. Wide variations in the remodeling response with regard to reduction in vessel diameter, intimal lesion formation, lumen area, and medial hypertrophy were found. On carotid artery ligation SJL/J mice revealed the most extensive inward remodeling leading to an approximate 78% decrease in lumen area while lumen narrowing in FVB/NJ mice was largely due to extensive neointima formation as a result of smooth muscle cell (SMC) proliferation. Significant positive remodeling in the contralateral right carotid artery with a >20% increase in lumen area was observed in SM/J and A/J mice. An in vitro comparison of growth properties of SMC isolated from FVB/NJ mice and a strain that exhibited very little SMC proliferation (C3H/HeJ) demonstrated accelerated growth of SMC from FVB/NJ following serum stimulation. In vivo, SMC proliferation in the FVB/NJ strain was preceded by a 37% loss of medial SMC occurring within the 2 days after ligation, however, cell death was not detectable in C3H/HeJ mice. These findings suggest that the mechanisms leading to lumen narrowing in the vascular remodeling process are genetically controlled. PMID- 10793087 TI - BRCA2 is inactivated late in the development of pancreatic intraepithelial neoplasia: evidence and implications. AB - Patients harboring germline BRCA2 mutations are at an increased risk of developing pancreatic cancer. We investigated the prevalence of biallelic inactivation of BRCA2 in the presumed precursors to invasive pancreatic ductal carcinomas, pancreatic intraepithelial neoplasia (PanIN). Surgical resection specimens from three patients with germline BRCA2 mutations who developed pancreatic ductal adenocarcinoma were studied. Fourteen PanINs were needle microdissected from paraffin-embedded tissue. DNA was isolated from these microdissected tissues and amplified by primer-mediated pre-amplification. Loss of heterozygosity at the BRCA2 locus was determined by polymerase chain reaction amplification and cycle sequencing. The presence of the wild-type alleles was evaluated at the nucleotide positions of the germline BRCA2 mutations. The K-ras gene was sequenced at codon 12 and 13 to confirm the efficacy of microdissection. By histological evaluation the prevalence of PanINs in these patients was not notably elevated. Loss of the wild-type allele of BRCA2 was present in one high grade PanIN (PanIN 3), but in none of 13 low-grade PanINs (PanIN 1). In contrast, K-ras mutations were detectable in 7 of the 14 PanINs. These results suggest that biallelic inactivation of the BRCA2 gene is a relatively late event in pancreatic tumorigenesis. In contrast to classical molecular progression models of tumorigenesis, the inactivation of the wild-type allele in a carrier of a recessive tumor susceptibility gene may not always be the first somatic event during the molecular evolution of a cancer. The necessity for earlier genetic alterations before biallelic inactivation of a recessive tumor susceptibility gene such as BRCA2 may explain why affected carriers have normal numbers of neoplastic precursor lesions, a relatively low phenotypic penetrance, and late age of onset of pancreatic and other cancers. PMID- 10793086 TI - Expression of agrin, dystroglycan, and utrophin in normal renal tissue and in experimental glomerulopathies. AB - The dystrophin-glycoprotein complex, which comprises alpha- and beta dystroglycan, sarcoglycans, and utrophin/dystrophin, links the cytoskeleton to agrin and laminin in the basal lamina in muscle and epithelial cells. Recently, agrin was identified as a major heparan sulfate proteoglycan in the glomerular basement membrane. In the present study, we found mRNA expression for agrin, dystroglycan, and utrophin in kidney cortex, isolated glomeruli, and cultured podocytes and mesangial cells. In immunofluorescence, agrin was found in the glomerular basement membrane. The antibodies against alpha- and beta-dystroglycan and utrophin revealed a granular podocyte-like staining pattern along the glomerular capillary wall. With immunoelectron microscopy, agrin was found in the glomerular basement membrane, dystroglycan was diffusely found over the entire cell surface of the podocytes, and utrophin was localized in the cytoplasm of the podocyte foot processes. In adriamycin nephropathy, a decrease in the glomerular capillary wall staining for dystroglycan was observed probably secondary to the extensive fusion of foot processes. Immunoelectron microscopy showed a different distribution pattern as compared to the normal kidney, with segmentally enhanced expression of dystroglycan at the basal side of the extensively fused podocyte foot processes. In passive Heymann nephritis we observed no changes in the staining intensity and distribution of the dystrophin-glycoprotein complex by immunofluorescence and immunoelectron microscopy. From these data, we conclude that agrin, dystroglycan, and utrophin are present in the glomerular capillary wall and their ultrastructural localization supports the concept that these molecules are involved in linking the podocyte cytoskeleton to the glomerular basement membrane. PMID- 10793088 TI - Genetic and epigenetic modification of MLH1 accounts for a major share of microsatellite-unstable colorectal cancers. AB - Microsatellite instability (MSI) is a hallmark of hereditary nonpolyposis colorectal cancer, and in these patients, results from inherited defects in DNA mismatch repair genes, mostly MSH2 and MLH1. MSI also occurs in 15% of sporadic colorectal cancers, but in these tumors, its basis is less well characterized. We investigated 46 sporadic MSI+ colorectal cancers for changes in MSH2 and MLH1 protein expression, followed by the analysis of somatic mutation, loss of heterozygosity (LOH), and promoter hypermethylation as possible underlying defects. Most cases (36/46, 78%) showed lost or reduced MLH1 expression. Among these, a majority (83%) was associated with MLH1 promoter hypermethylation, whereas the rates of LOH and somatic mutation of MLH1 were 24% and 13%, respectively. Hypermethylation and LOH were inversely correlated, suggesting that they had alternative functions in the inactivation of MLH1. MSH2 expression was lost in 7/46 (15%), and of these, 2 (29%) showed LOH and/or somatic mutation of MSH2. We conclude that most sporadic MSI+ colorectal cancers have an MLH1 associated etiology and that epigenetic modification is a major mechanism of MLH1 inactivation. Moreover, we found a significantly lower prevalence for MLH1 promoter hypermethylation in hereditary nonpolyposis colorectal cancer tumors with MLH1 germline mutations (12/26, 46%), which might explain some differences that are known to occur in the clinicopathological characteristics and tumorigenic pathways between sporadic and hereditary MSI+ colorectal cancers. PMID- 10793089 TI - Reduced leptin levels in starvation increase susceptibility to endotoxic shock. AB - Malnutrition compromises immune function, reducing resistance to infection. We examine whether the decrease in leptin induced by starvation increases susceptibility to lipopolysaccharide (LPS)- and tumor necrosis factor (TNF) induced lethality. In mice, fasting for 48 hours enhances sensitivity to LPS. Decreasing the fasting-induced fall in leptin by leptin administration markedly reduced sensitivity to LPS. Although fasting decreases basal leptin levels, LPS treatment increased leptin to the same extent as in fed animals. Fasting increased basal serum corticosterone; leptin treatment blunted this increase. Fasting decreased the ability of LPS to increase corticosterone; leptin restored the corticosterone response to LPS. Serum glucose levels were decreased in fasted mice and LPS induced a further decrease. Leptin treatment affected neither basal glucose nor that after LPS. LPS induced a fivefold greater increase in serum TNF in fasted mice, which was blunted by leptin replacement. In contrast, LPS induced lower levels of interferon-gamma and no differences in interleukin-1beta in fasted compared to fed animals; leptin had no effect on those cytokines. Furthermore, fasting increased sensitivity to the lethal effect of TNF itself, which was also reversed by leptin treatment. Thus, leptin seems to be protective by both inhibiting TNF induction by LPS and by reducing TNF toxicity. PMID- 10793090 TI - Eosinophilia of dystrophin-deficient muscle is promoted by perforin-mediated cytotoxicity by T cell effectors. AB - Previous investigations have shown that cytotoxic T lymphocytes (CTLs) contribute to muscle pathology in the dystrophin-null mutant mouse (mdx) model of Duchenne muscular dystrophy through perforin-dependent and perforin-independent mechanisms. We have assessed whether the CTL-mediated pathology includes the promotion of eosinophilia in dystrophic muscle, and thereby provides a secondary mechanism through which CTLs contribute to muscular dystrophy. Quantitative immunohistochemistry confirmed that eosinophilia is a component of the mdx dystrophy. In addition, electron microscopic observations show that eosinophils traverse the basement membrane of mdx muscle fibers and display sites of close apposition of eosinophil and muscle membranes. The close membrane apposition is characterized by impingement of eosinophilic rods of major basic protein into the muscle cell membrane. Transfer of mdx splenocytes and mdx muscle extracts to irradiated C57 mice by intraperitoneal injection resulted in muscle eosinophilia in the recipient mice. Double-mutant mice lacking dystrophin and perforin showed less eosinophilia than was displayed by mdx mice that expressed perforin. Finally, administration of prednisolone, which has been shown previously to reduce the concentration of CTLs in dystrophic muscle, produced a significant reduction in eosinophilia. These findings indicate that eosinophilia is a component of the mdx pathology that is promoted by perforin-dependent cytotoxicity of effector T cells. However, some eosinophilia of mdx muscle is independent of perforin-mediated processes. PMID- 10793091 TI - Biomechanical regulation of human monocyte/macrophage molecular function. AB - When the monocyte infiltrates a tissue, adhesion to the extracellular matrix provides structural anchors, and the cell may be deformed through these attachments. To test the hypothesis that human monocytes/macrophages are mechanically responsive, we studied the effects of small cyclic mechanical deformations on cultured human monocytes/macrophages. When monocytes/macrophages were subjected to 4% strain at 1 Hz for 24 hours, neither matrix metalloproteinase (MMP)-1 nor MMP-3 was induced; however, in the presence of phorbol myristate acetate, strain augmented MMP-1 expression by 5.1 +/- 0.7-fold (P < 0.05) and MMP-3 expression by 1. 6 +/- 0.1-fold (P < 0.05). In contrast, MMP 9 expression was not changed by mechanical strain in the presence or absence of phorbol myristate acetate. Deformation rapidly induced the immediate early response genes c-fos and c-jun. In addition, mechanical deformation induced the transcription factor PU.1, an ets family member that is essential in monocyte differentiation, as well as mRNA for the M-CSF receptor. These studies demonstrate that human monocytes/macrophages respond to mechanical deformation with selective augmentation of MMPs, induction of immediate early genes, and induction of the M-CSF receptor. In addition to enhancing the proteolytic activity of macrophages within repairing tissues, cellular deformation within tissues may play a role in monocyte differentiation. PMID- 10793092 TI - Genetic characterization of Trypanosoma cruzi directly from tissues of patients with chronic Chagas disease: differential distribution of genetic types into diverse organs. AB - We have previously shown that a low-stringency single-specific primer-polymerase chain reaction (LSSP- PCR) is a highly sensitive and reproducible technique for the genetic profiling of Trypanosoma cruzi parasites directly in tissues from infected animals and humans. By applying LSSP-PCR to the study of the variable region of kinetoplast minicircle from T. cruzi, the intraspecific polymorphism of the kinetoplast-deoxyribonucleic acid (kDNA) sequence can be translated into individual kDNA signatures. In the present article, we report on our success using the LSSP-PCR technique in profiling the T. cruzi parasites present in the hearts of 13 patients with chagasic cardiopathy and in the esophagi of four patients (three of them with chagasic megaesophagus). In two patients, one with the cardiodigestive clinical form of Chagas disease and the other with cardiopathy and an esophageal inflammatory process, we could study both heart and esophagus and we detected distinct kDNA signatures in the two organs. This provides evidence of a differential tissue distribution of genetically diverse T. cruzi populations in chronic Chagas disease, suggesting that the genetic variability of the parasite is one of the determining factors of the clinical form of the disease. PMID- 10793094 TI - Confirmation of the "old" standard of care for ovarian cancer and a challenge. PMID- 10793093 TI - Interferon-gamma and tumor necrosis factor-alpha determine resistance to Paracoccidioides brasiliensis infection in mice. AB - To investigate the role of interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in the resistance to Paracoccidioides brasiliensis (Pb) infection, mice with homologous disruption of the IFN-gamma (GKO) or TNF-alpha receptor p55 (p55KO) were infected with the parasite. GKO and p55KO, but not wild type (WT) mice, were unable to control the growth of yeast cells and the mice succumbed to infection by days 16 and 90 after infection, respectively. Typical inflammatory granulomas were found only in WT mice. In contrast, knockout mice presented an inflammatory infiltrate composed of a few neutrophils, mononuclear, epithelioid, and multinuclear giant cells forming incipient granulomas in GKO mice and without granuloma formation in p55KO mice. Besides, both groups of knockout mice exhibited elevated numbers of yeast forms in agreement with colony forming unit counts in organs. Compared with WT, splenocytes from infected GKO mice cultured with the Pb F1 fraction produced lower TNF-alpha levels, whereas leukocytes from infected p55KO mice produced similar amounts of TNF-alpha but higher levels of IFN-gamma. Moreover, splenocytes from infected WT mice produced higher levels of nitric oxide (NO) resulting in a lower T-cell proliferative response to Con A than uninfected WT, or infected p55KO and GKO mice. On the contrary, the addition of IFN-gamma to splenocytes from infected GKO mice resulted in higher NO production and lower T cell proliferation. Taken together, these findings suggests that endogenous TNF-alpha, acting through the p55 receptor, and IFN-gamma mediate resistance to Pb infection and induce NO production that determines marked T cell unresponsiveness. PMID- 10793095 TI - Association of human papillomaviruses with a subgroup of head and neck squamous cell carcinomas. PMID- 10793097 TI - HPV in oropharyngeal cancers: new data inspire hope for vaccines. PMID- 10793096 TI - Addressing controversies over Kaposi's sarcoma. PMID- 10793098 TI - Multinational effort aims to record all clinical trials. PMID- 10793099 TI - Eliminating publication bias: the effect of negative trial results. PMID- 10793101 TI - Brain tumor research facility debuts in Montreal. PMID- 10793100 TI - Herceptin in the adjuvant setting: phase III trials begin. PMID- 10793102 TI - Researchers search for link between circadian rhythms, breast cancer. PMID- 10793103 TI - Stat bite: All-cancer mortality in selected countries in the Americas, 1995. PMID- 10793104 TI - Children's inn celebrates 10(th) anniversary PMID- 10793105 TI - Papillomaviruses causing cancer: evasion from host-cell control in early events in carcinogenesis. AB - During the past 20 years, several types of human papillomaviruses (HPVs) have been identified that cause specific types of cancers. The etiology of cancer of the cervix has been linked to several types of HPV, with a high preponderance of HPV16. The role of these virus infections has been established 1) by the regular presence of HPV DNA in the respective tumor biopsy specimens, 2) by the demonstration of viral oncogene expression (E6 and E7) in tumor material, 3) by the identification of transforming properties of these genes, 4) by the requirement for E6 and E7 expression for maintaining the malignant phenotype of cervical carcinoma cell lines, 5) by the interaction of viral oncoproteins with growth-regulating host-cell proteins, and 6) by epidemiologic studies pointing to these HPV infections as the major risk factor for cervical cancer development. In addition to cancer of the cervix, a major proportion of anal, perianal, vulvar, and penile cancers appears to be linked to the same HPV infections. In addition, close to 20% of oropharyngeal cancers contain DNA from the same types of HPV. Recent evidence also points to a possible role of other HPV infections in squamous cell carcinomas of the skin. This review covers recent developments in understanding molecular mechanisms of HPV carcinogenesis, mainly discussing functions of viral oncoproteins and the regulation of viral oncogenes by host cell factors. Modifications in host-cell genes, most likely engaged in the control of HPV gene expression in proliferating cells, emerge as important events in HPV-mediated carcinogenesis. PMID- 10793106 TI - Randomized intergroup trial of cisplatin-paclitaxel versus cisplatin cyclophosphamide in women with advanced epithelial ovarian cancer: three-year results. AB - BACKGROUND: A randomized trial conducted by the Gynecologic Oncology Group (GOG, study #111) in the United States showed a better outcome for patients with advanced ovarian cancer on the paclitaxel-cisplatin regimen than for those on a standard cyclophosphamide-cisplatin regimen. Before considering the paclitaxel cisplatin regimen as the new "standard," a group of European and Canadian investigators planned a confirmatory phase III trial. METHODS: This intergroup trial recruited 680 patients with broader selection criteria than the GOG #111 study and administered paclitaxel as a 3-hour instead of a 24-hour infusion; progression-free survival was the primary end point. Patient survival was analyzed by use of the Kaplan-Meier technique. Treatment effects on patient survival were estimated by Cox proportional hazards regression models. All statistical tests were two-sided. RESULTS: The overall clinical response rate was 59% in the paclitaxel group and 45% in the cyclophosphamide group; the complete clinical remission rates were 41% and 27%, respectively; both differences were statistically significant (P =.01 for both). At a median follow-up of 38.5 months and despite a high rate of crossover (48%) from the cyclophosphamide arm to the paclitaxel arm at first detection of progression of disease, a longer progression free survival (log-rank P =.0005; median of 15.5 months versus 11.5 months) and a longer overall survival (log-rank P =. 0016; median of 35.6 months versus 25.8 months) were seen in the paclitaxel regimen compared with the cyclophosphamide regimen. CONCLUSIONS: There is strong and confirmatory evidence from two large randomized phase III trials to support paclitaxel-cisplatin as the new standard regimen for treatment of patients with advanced ovarian cancer. PMID- 10793107 TI - Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. AB - BACKGROUND: High-risk human papillomaviruses (HPVs) are etiologic agents for anogenital tract cancers and have been detected in head and neck squamous cell carcinomas (HNSCCs). We investigated, retrospectively, an etiologic role for HPVs in a large series of patients with HNSCC. METHODS: Tumor tissues from 253 patients with newly diagnosed or recurrent HNSCC were tested for the presence of HPV genome by use of polymerase chain reaction (PCR)-based assays, Southern blot hybridization, and in situ hybridization. The viral E6 coding region was sequenced to confirm the presence of tumor-specific viral isolates. Exons 5-9 of the TP53 gene were sequenced from 166 specimens. The hazard of death from HNSCC in patients with and without HPV-positive tumors was determined by proportional hazards regression analysis. RESULTS: HPV was detected in 62 (25%) of 253 cases (95% confidence interval [CI] = 19%-30%). High-risk, tumorigenic type HPV16 was identified in 90% of the HPV-positive tumors. HPV16 was localized specifically by in situ hybridization within the nuclei of cancer cells in preinvasive, invasive, and lymph node disease. Southern blot hybridization patterns were consistent with viral integration. Poor tumor grade (odds ratio [OR] = 2.4; 95% CI = 1.2- 4.9) and oropharyngeal site (OR = 6.2; 95% CI = 3.1-12.1) independently increased the probability of HPV presence. As compared with HPV-negative oropharyngeal cancers, HPV-positive oropharyngeal cancers were less likely to occur among moderate to heavy drinkers (OR = 0.17; 95% CI = 0.05-0.61) and smokers (OR = 0.16; 95% CI = 0.02-1.4), had a characteristic basaloid morphology (OR = 18.7; 95% CI = 2.1 167), were less likely to have TP53 mutations (OR = 0.06; 95% CI = 0.01-0. 36), and had improved disease-specific survival (hazard ratio [HR] = 0.26; 95% CI = 0.07-0.98). After adjustment for the presence of lymph node disease (HR = 2.3; 95% CI = 1.4- 3.8), heavy alcohol consumption (HR = 2.6; 95% CI = 1.4-4.7), and age greater than 60 years old (HR = 1.4; 95% CI = 0.8-2.3), all patients with HPV positive tumors had a 59% reduction in risk of death from cancer when compared with HPV-negative HNSCC patients (HR = 0.41; 95% CI = 0.20-0.88). CONCLUSIONS: These data extend recent molecular and epidemiologic studies and strongly suggest that HPV-positive oropharyngeal cancers comprise a distinct molecular, clinical, and pathologic disease entity that is likely causally associated with HPV infection and that has a markedly improved prognosis. PMID- 10793108 TI - Human Kaposi's sarcoma cell-mediated tumorigenesis in human immunodeficiency type 1 tat-expressing transgenic mice. AB - BACKGROUND: The human immunodeficiency virus type 1 (HIV-1) transactivator (Tat) protein has been linked to the development and course of Kaposi's sarcoma (KS) associated with acquired immunodeficiency disease syndrome (AIDS-KS). Tat is an 86-101 amino-acid protein encoded by two exons. To evaluate the growth-promoting effects of Tat in AIDS-KS in vivo, we developed transgenic mice expressing the one-exon-encoded 72 amino-acid protein (Tat(72)) and the two-exon-encoded 86 amino-acid protein (Tat(86)). METHODS: Human KS SLK cells were injected subcutaneously into CD4(+) T-cell-depleted male mice, and the tumors that formed after 3-4 weeks were recovered and analyzed for the expression of Tat protein(s), different cytokine messenger RNAs (mRNAs), and matrix metalloproteinases (MMPs). All statistical tests were two-sided. RESULTS: The average tumor weight was maximum in Tat(86) mice ( approximately 600 mg) compared with Tat(72) ( approximately 200 mg) and nontransgenic ( approximately 100 mg) mice (P<.005). Histologic examination of tumors showed spindle-shaped SLK cells with prominent infiltrates of inflammatory cells. All of the tumors from Tat mice expressed abundant Tat mRNA, suggesting that the infiltrating mouse cells actively expressed Tat. A comparison of the growth-promoting cytokines in the tumors from Tat(86)-transgenic and nontransgenic mice showed that the expression of the following cytokines was substantially increased in the tumors of the Tat(86) mice: tumor necrosis factor-alpha, interleukin 6, interleukin 8, granulocyte macrophage colony-stimulating factor, and basic fibroblast growth factor. Furthermore, these tumors showed abundant expression of a 105-kd MMP activity associated with infiltrates of host leukocytes in the lesions. CONCLUSION: Our in vivo data clearly suggest that extracellular Tat can contribute to the growth and tumorigenesis of human KS cells. PMID- 10793109 TI - Monoclonality or oligoclonality of human herpesvirus 8 terminal repeat sequences in Kaposi's sarcoma and other diseases. AB - BACKGROUND: Infection with human herpesvirus 8 (HHV8), also termed Kaposi's sarcoma (KS)-associated herpesvirus, is associated with all forms of KS, with primary effusion lymphoma (PEL), and with some forms of multicentric Castleman's disease (MCD), but the pathogenic role of HHV8 in these tumors and the clonal nature of KS are still unclear. The purpose of this study was to examine whether the number of terminal repeats (TRs) contained in the fused TR region of HHV8 could be used as a marker of clonality in HHV8-associated tumors. METHODS: Pulsed field gel electrophoresis (PFGE) and multiple-probe Southern blot analysis of the HHV8 TR region were performed on high-molecular-weight DNA obtained from tumoral KS, PEL, and MCD lesions. RESULTS: These analysis showed that the fused TR region contains a large but variable number of TR units (ranging from 16 to 75) and that the viral genome is present as extrachromosomal circular DNA in these tumors in vivo, with occasional ladders of heterogeneous linear termini reflecting lytic replication. All PEL tumors and PEL-derived cell lines as well as some KS tumors contained monoclonal or oligoclonal fused TR fragments; however, the TR region appeared polyclonal in MCD tumors and in a few KS lesions. CONCLUSION: Several KS and PEL lesions are monoclonal expansions of a single infected cell, suggesting that HHV8 infection precedes tumor growth and thus supporting an etiologic role of latent HHV8 in these proliferations. Our finding that nodular KS lesions display all possible patterns of clonality supports the model according to which KS begins as a polyclonal disease with subsequent evolution to a monoclonal process. PMID- 10793111 TI - Biologic characteristics of interval and screen-detected breast cancers. AB - BACKGROUND: Interval breast cancer is defined as a cancer that is detected within 12 months after a negative mammogram. The failure of mammography to detect breast cancer depends on testing procedures, radiologist interpretation, patient characteristics, and tumor properties. Although errors by radiologists explain some interval cancers, another explanation is that the tumor is rapidly growing and was too small to be detected on the last mammogram. To determine whether markers of tumor growth rate are associated with risk of an interval cancer, we conducted a population-based study with the use of data collected statewide by the New Mexico Mammography Project. METHODS: Among women who received a mammographic examination from 1991 throughout 1993, we ascertained records of all patients with breast cancer diagnosed within 12 months of a negative screening mammographic examination (interval cancers) and corresponding tumor samples, when available. We selected an age- and ethnicity-matched comparison group of control patients with screen-detected breast cancers diagnosed during the same period. In tumor samples, p53, bcl-2, and Ki-67 were examined immunologically and the apoptotic index was assessed histologically. We used logistic regression to determine whether interval cancers were associated with selected demographic, radiologic, and biologic characteristics. RESULTS: It is more likely that mammography did not detect tumors with a high proportion of proliferating cells (>20%) than tumors with a low proportion of proliferating cells (<5%) (odds ratio [OR] = 4.09; 95% confidence interval [CI] = 1.14-14.65). The OR for mammographic failure was 2.96 (95% CI = 1.07-8.20) among cancers that expressed p53 compared with cancers that did not. Interval cancers also had fewer apoptotic cells. Approximately 75% of interval cancers appear to have tumors with 5% proliferating cells or more. Younger women had a higher proportion of rapidly proliferating and aggressive cancers. CONCLUSION: Rapidly growing and aggressive tumors account for a substantial proportion of mammographic failure to detect breast cancer, especially among younger women, who have a high proportion of aggressive cancers. PMID- 10793110 TI - Joint effect of insulin-like growth factors and mutagen sensitivity in lung cancer risk. AB - BACKGROUND: We hypothesize that accumulation of genetic damage is dependent on an individual's intrinsic carcinogen sensitivity and on various humoral factors (e.g., insulin-like growth factors [IGFs]) that enhance proliferation, resistance to apoptotic cell death, and clonal outgrowth of genetically damaged cells. We tested this hypothesis by determining whether proliferation potential and genetic instability are associated with the risk of lung cancer. METHODS: In a study of 183 lung cancer patients and 227 matched control subjects, we examined the joint effects of latent genetic instability (measured as mutagen sensitivity) and elevated proliferation potential (assessed by measuring IGFs) in lung cancer risk. Levels of IGF-I, IGF-II, and IGF-binding protein-3 (IGFBP-3) in plasma were measured by use of immunoassay kits. Mutagen sensitivity was assessed by quantitating bleomycin- and benzo[a]pyrene diol epoxide (BPDE)-induced chromatid breaks in peripheral blood lymphocyte cultures. RESULTS: Although not statistically significant, the mean levels of IGF-I and the molar ratio of IGF I/IGFBP-3 were higher in patients with advanced or poorly differentiated disease than in patients with early or well-differentiated disease. Variation in IGFs was not associated with any specific histologic type or tumor stage. High levels of IGF-I and enhanced mutagen sensitivity were individually associated with increased risk of lung cancer: odds ratio (OR) of 2.13 (95% confidence interval [CI] = 1.20-3.78) for IGF-I, 2.50 (95% CI = 1. 49-4.20) for bleomycin sensitivity, and 2.95 (95% CI = 1.72-5.06) for BPDE sensitivity. The OR was statistically significantly elevated to 8.88 for both higher IGF-I and bleomycin sensitivity (95% CI = 3.67-21.50) and to 13.53 for higher IGF-I and BPDE sensitivity combined (95% CI = 4.48-40.89). With all three risk factors considered together, the OR was 17.09 (95% CI = 4.16-70.27). High levels of IGFBP 3 alone were associated with reduced lung cancer risk: OR = 0.59 (95% CI = 0.33 1.05). CONCLUSIONS: Our data suggest that individuals with genetic instability and higher proliferation potential are at enhanced risk for lung cancer. PMID- 10793112 TI - Organochlorine pesticide content of breast adipose tissue from women with breast cancer and control subjects. PMID- 10793113 TI - Re: HRAS1 rare minisatellite alleles and breast cancer in Australian women under age forty years. PMID- 10793114 TI - Unexpected right phrenic nerve injury during 5-fluorouracil continuous infusion plus cisplatin and vinorelbine in breast cancer patients. PMID- 10793115 TI - Re: HRAS1 rare minisatellite alleles and breast cancer in australian women under age forty years PMID- 10793116 TI - RESPONSE: re: HRAS1 rare minisatellite alleles and breast cancer in australian women under age forty years PMID- 10793117 TI - Re: weighing the risks and benefits of tamoxifen treatment for preventing breast cancer. PMID- 10793118 TI - Re: biologic and biochemical analyses of p16(INK4a) mutations from primary tumors. PMID- 10793119 TI - RESPONSE: re: weighing the risks and benefits of tamoxifen treatment for preventing breast cancer PMID- 10793121 TI - RESPONSE: re: biologic and biochemical analyses of p16(INK4a) mutations from primary tumors PMID- 10793120 TI - Re: biologic and biochemical analyses of p16(INK4a) mutations from primary tumors PMID- 10793122 TI - RESPONSE: re: environmental tobacco smoke, genetic susceptibility, and risk of lung cancer in never-smoking women PMID- 10793123 TI - Re: environmental tobacco smoke, genetic susceptibility, and risk of lung cancer in never-smoking women. PMID- 10793124 TI - RESPONSE: re: environmental tobacco smoke, genetic susceptibility, and risk of lung cancer in never-smoking women PMID- 10793125 TI - Characteristics of empirically supported treatments. AB - This study presents a survey of general characteristics of empirically supported treatments (ESTs) identified by the American Psychological Association Division 12 Task Force on the Promotion and Dissemination of Psychological Procedures. Results indicate that the ESTs share the following characteristics: they involve skill building, have a specific problem focus, incorporate continuous assessment of client progress, and involve brief treatment contact, requiring 20 or fewer sessions. Traditional assessment methods, such as intelligence testing, projectives, and objective personality tests such as the MMPI-2, are rarely used in these treatments. Although it is recognized that these findings are in part an artifact of sociological factors present in contemporary psychotherapy development and research, the findings may also serve as a heuristic aid in the development of therapies. PMID- 10793126 TI - Psychotherapy adherence of therapists treating HIV-positive patients with depressive symptoms. AB - Adherence monitoring, a technology to specify research psychotherapies, was used in the NIMH Treatment of Depression Collaborative Research Program (TDCRP). The authors present adherence data from a similar randomized treatment trial of 56 depressed HIV-positive patients, comparing 16-week interventions with cognitive behavioral therapy, interpersonal psychotherapy, and supportive psychotherapy alone or with imipramine. Therapists were certified in manualized treatments. Blind independent raters rated randomly selected taped sessions on an adaptation of the NIMH scale, yielding adherence scores for interventions and for therapist "facilitative conditions" (FC). All therapists were rated adherent. Interrater reliability was 0.89-0.99. The scale discriminated among the four treatments (P<0.0001), with each scoring highest on its own scale. FC, which might measure therapist competence independent of treatment technique, varied by intervention but did not predict treatment outcome. This study demonstrates the ability to reliably train adherence monitors and therapists able to deliver specified treatments. Its adherence findings provide the first replication of those from the landmark NIMH TDCRP study. PMID- 10793128 TI - Time-limited psychotherapy with adolescents. AB - Short-term dynamic therapies, characterized by abbreviated lengths (10-40 sessions) and, in many cases, preset termination dates, have become more widespread in the past three decades. Short-term therapies are based on rapid psychodynamic diagnosis, a therapeutic focus, a rapidly formed therapeutic alliance, awareness of termination and separation processes, and the directive stance of the therapist. The emotional storm of adolescence, stemming from both developmental and psychopathological sources, leaves many adolescents in need of psychotherapy. Many adolescents in need of therapy resist long-term attachment and involvement in an ambiguous relationship, which they experience as a threat to their emerging sense of independence and separateness. Short-term dynamic therapy can be the treatment of choice for many adolescents because it minimizes these threats and is more responsive to their developmental needs. The article presents treatment and follow-up of a 17-year-old youth, using James Mann's time limited psychotherapy method. PMID- 10793127 TI - The use of dreams in psychotherapy: a survey of psychotherapists in private practice. AB - Since the publication of Sigmund Freud's The Interpretation of Dreams, dream interpretation has been a standard technique often used in psychotherapy. However, empirical studies about the frequency of working on dreams in therapy are lacking. The present study elicited, via a self-developed questionnaire, various aspects of work on dreams applied by psychotherapists in private practice. The findings indicate that dreams were often used in therapy, especially in psychoanalysis. In addition, a significant relationship was found between the frequency of the therapists' working on their own dreams and frequency of work on dreams in therapy. Because work on dreams was rated as beneficial for the clients, further studies investigating the effectiveness and the process of working on dreams will be of interest. PMID- 10793129 TI - Can Therapy Affect Physical Health? PMID- 10793130 TI - The microfibrillar proteins MAGP-1 and fibrillin-1 form a ternary complex with the chondroitin sulfate proteoglycan decorin. AB - MAGP-1 and fibrillin-1, two protein components of extracellular microfibrils, were shown by immunoprecipitation studies to interact with the chondroitin sulfate proteoglycan decorin in the medium of cultured fetal bovine chondrocytes. Decorin interacted with each protein individually and with both proteins together to form a ternary complex. Expression of truncated fibrillin-1 proteins in Chinese hamster ovary cells localized proteoglycan binding to an amino-terminal region near the proline-rich domain. A spatially analogous fibrillin-2 truncated protein did not coprecipitate the same sulfated molecule, suggesting that chondroitin sulfate proteoglycan binding in this region is specific for fibrillin 1. An interaction between fibrillin and MAGP-1 was also observed under culture conditions that abrogated decorin secretion, suggesting that the two microfibrillar proteins can associate in the absence of the proteoglycan. Sulfation of matrix proteins is important for elastic fiber assembly because inhibition of sulfation was shown to prevent microfibrillar protein incorporation into the extracellular matrix of cultured cells. PMID- 10793131 TI - Identification of a novel member of the chloride intracellular channel gene family (CLIC5) that associates with the actin cytoskeleton of placental microvilli. AB - The chloride intracellular channel (CLIC) gene family has been implicated in chloride ion transport within various subcellular compartments. We report here the molecular, biochemical, and cellular characterization of a new member of this gene family termed CLIC5. CLIC5 was isolated from extracts of placental microvilli as a component of a multimeric complex consisting of several known cytoskeletal proteins, including actin, ezrin, alpha-actinin, gelsolin, and IQGAP1. We cloned human cDNAs and generated antibodies specific for CLIC5, CLIC1/NCC27, and CLIC4/huH1/p64H1. CLIC5 shares 52-76% overall identity with human CLIC1, CLIC2, CLIC3, and CLIC4. Northern blot analysis showed that CLIC5 has a distinct pattern of expression compared with CLIC1 and CLIC4. Immunoblot analysis of extracts from placental tissues demonstrated that CLIC4 and CLIC5 are enriched in isolated placental microvilli, whereas CLIC1 is not. Moreover, in contrast to CLIC1 and CLIC4, CLIC5 is associated with the detergent-insoluble cytoskeletal fraction of microvilli. Indirect immunofluorescence microscopy revealed that CLIC4 and CLIC5 are concentrated within the apical region of the trophoblast, whereas CLIC1 is distributed throughout the cytoplasm. These studies suggest that CLIC1, CLIC4, and CLIC5 play distinct roles in chloride transport and that CLIC5 interacts with the cortical actin cytoskeleton in polarized epithelial cells. PMID- 10793132 TI - Gaa1p and gpi8p are components of a glycosylphosphatidylinositol (GPI) transamidase that mediates attachment of GPI to proteins. AB - Many eukaryotic cell surface proteins are anchored to the membrane via glycosylphosphatidylinositol (GPI). The GPI is attached to proteins that have a GPI attachment signal peptide at the carboxyl terminus. The GPI attachment signal peptide is replaced by a preassembled GPI in the endoplasmic reticulum by a transamidation reaction through the formation of a carbonyl intermediate. GPI transamidase is a key enzyme of this posttranslational modification. Here we report that Gaa1p and Gpi8p are components of a GPI transamidase. To determine a role of Gaa1p we disrupted a GAA1/GPAA1 gene in mouse F9 cells by homologous recombination. GAA1 knockout cells were defective in the formation of carbonyl intermediates between precursor proteins and transamidase as determined by an in vitro GPI-anchoring assay. We also show that cysteine and histidine residues of Gpi8p, which are conserved in members of a cysteine protease family, are essential for generation of a carbonyl intermediate. This result suggests that Gpi8p is a catalytic component that cleaves the GPI attachment signal peptide. Moreover, Gaa1p and Gpi8p are associated with each other. Therefore, Gaa1p and Gpi8p constitute a GPI transamidase and cooperate in generating a carbonyl intermediate, a prerequisite for GPI attachment. PMID- 10793133 TI - Response of Xenopus Cds1 in cell-free extracts to DNA templates with double stranded ends. AB - Although homologues of the yeast checkpoint kinases Cds1 and Chk1 have been identified in various systems, the respective roles of these kinases in the responses to damaged and/or unreplicated DNA in vertebrates have not been delineated precisely. Likewise, it is largely unknown how damaged DNA and unreplicated DNA trigger the pathways that contain these effector kinases. We report that Xenopus Cds1 (Xcds1) is phosphorylated and activated by the presence of some simple DNA molecules with double-stranded ends in cell-free Xenopus egg extracts. Xcds1 is not affected by aphidicolin, an agent that induces DNA replication blocks. In contrast, Xenopus Chk1 (Xchk1) responds to DNA replication blocks but not to the presence of double-stranded DNA ends. Immunodepletion of Xcds1 (and/or Xchk1) from egg extracts did not attenuate the cell cycle delay induced by double-stranded DNA ends. These results imply that the cell cycle delay triggered by double-stranded DNA ends either does not involve Xcds1 or uses a factor(s) that can act redundantly with Xcds1. PMID- 10793134 TI - Nuclear matrix-like filaments and fibrogranular complexes form through the rearrangement of specific nuclear ribonucleoproteins. AB - The behavior of nuclear pre-mRNA-binding proteins after their nuclease and/or salt-induced release from RNA was investigated. After RNase digestion or salt extraction, two proteins that initially exist as tetramers (A2)(3)B1 in isolated heterogeneous nuclear ribonucleoprotein (hnRNP) complexes quantitatively reassociated to form regular helical filaments ranging in length from 100 nm to >10 microm. In highly magnified preparations prepared for scanning transmission electron microscopy, single filaments have diameters near 18 nm. In conventional negatively stained preparations viewed at low magnification, the diameters of the thinnest filaments range from 7 to 10 nm. At protein concentrations of >0.1 mg/ml, the filaments rapidly aggregated to form thicker filamentous networks that look like the fibrogranular structures termed the "nuclear matrix." Like the residual material seen in nuclear matrix preparations, the hnRNP filaments were insoluble in 2 M NaCl. Filament formation is associated with, and may be dependent on, disulfide bridge formation between the hnRNP proteins. The reducing agent 2-mercaptoethanol significantly attenuates filament assembly, and the residual material that forms is ultrastructurally distinct from the 7- to 10-nm fibers. In addition to the protein rearrangement leading to filament formation, nearly one-third of the protein present in chromatin-clarified nuclear extracts was converted to salt-insoluble material within 1 min of digestion with RNase. These observations are consistent with the possibility that the residual material termed the nuclear matrix may be enriched in, if not formed by, denatured proteins that function in pre-mRNA packaging, processing, and transport. PMID- 10793135 TI - Mitotic regulation of the APC activator proteins CDC20 and CDH1. AB - The ordered activation of the ubiquitin protein ligase anaphase-promoting complex (APC) or cyclosome by CDC20 in metaphase and by CDH1 in telophase is essential for anaphase and for exit from mitosis, respectively. Here, we show that CDC20 can only bind to and activate the mitotically phosphorylated form of the Xenopus and the human APC in vitro. In contrast, the analysis of phosphorylated and nonphosphorylated forms of CDC20 suggests that CDC20 phosphorylation is neither sufficient nor required for APC activation. On the basis of these results and the observation that APC phosphorylation correlates with APC activation in vivo, we propose that mitotic APC phosphorylation is an important mechanism that controls the proper timing of APC(CDC20) activation. We further show that CDH1 is phosphorylated in vivo during S, G2, and M phase and that CDH1 levels fluctuate during the cell cycle. In vitro, phosphorylated CDH1 neither binds to nor activates the APC as efficiently as does nonphosphorylated CDH1. Nonphosphorylatable CDH1 mutants constitutively activate APC in vitro and in vivo, whereas mutants mimicking the phosphorylated form of CDH1 are constitutively inactive. These results suggest that mitotic kinases have antagonistic roles in regulating APC(CDC20) and APC(CDH1); the phosphorylation of APC subunits is required to allow APC activation by CDC20, whereas the phosphorylation of CDH1 prevents activation of the APC by CDH1. These mechanisms can explain the temporal order of APC activation by CDC20 and CDH1 and may help to ensure that exit from mitosis is not initiated before anaphase has occurred. PMID- 10793136 TI - Ca(2+) entry through store-operated channels in mouse sperm is initiated by egg ZP3 and drives the acrosome reaction. AB - Fertilization occurs after the completion of the sperm acrosome reaction, a secretory event that is triggered during gamete adhesion. ZP3, an egg zona pellucida glycoprotein, produces a sustained increase of the internal Ca(2+) concentration in mouse sperm, leading to acrosome reactions. Here we show that the sustained Ca(2+) concentration increase is due to the persistent activation of a Ca(2+) influx mechanism during the late stages of ZP3 signal transduction. These cells also possess a Ca(2+) store depletion-activated Ca(2+) entry pathway that is open after treatment with thapsigargin. Thapsigargin and ZP3 activate the same Ca(2+) permeation mechanism, as demonstrated by fluorescence quenching experiments and by channel antagonists. These studies show that ZP3 generates a sustained Ca(2+) influx through a store depletion-operated pathway and that this drives the exocytotic acrosome reaction. PMID- 10793137 TI - Hsp90 is essential for the synthesis and subsequent membrane association, but not the maintenance, of the Src-kinase p56(lck). AB - Tyrosine kinases of the Src family are synthesized as cytosolic proteins that subsequently translocate to membranes. Little is known of the mechanisms responsible for targeting these proteins to membranes, although a role for the cytosolic chaperone Hsp90 has been proposed. Here, we have studied the involvement of Hsp90 in the synthesis, membrane binding, and maintenance of the Src-kinase Lck. Using specific inhibitors of Hsp90, geldanamycin and radicicol, we found that functional Hsp90 is essential for the stability of newly synthesized, but not mature, Lck. Similar results were obtained for two other Src kinases, c-Src and Lyn. In contrast, LckY505F and LckDeltaSH2, constitutively active Lck mutants lacking the C-terminal regulatory tyrosine or the entire Src homology 2 domain, respectively, required Hsp90 activity to stabilize the mature proteins. Lck synthesized in the absence of Hsp90 activity was degraded within 30 45 min. This unstable Lck was myristoylated normally but did not associate with membranes or CD4, interactions that normally start within minutes of the completion of Lck synthesis. A construct composed of the N-terminal unique domain of Lck fused to green fluorescent protein did not require Hsp90 activity during synthesis. In addition, this protein associated with membranes efficiently in the absence of Hsp90 activity. Together these data suggest that interaction with Hsp90 is necessary for the correct synthesis and subsequent membrane binding of Lck. However, Hsp90 does not appear to play a direct role in Lck membrane, or CD4, association. PMID- 10793138 TI - Activating phosphorylation of the Saccharomyces cerevisiae cyclin-dependent kinase, cdc28p, precedes cyclin binding. AB - Eukaryotic cell cycle progression is controlled by a family of protein kinases known as cyclin-dependent kinases (Cdks). Two steps are essential for Cdk activation: binding of a cyclin and phosphorylation on a conserved threonine residue by the Cdk-activating kinase (CAK). We have studied the interplay between these regulatory mechanisms during the activation of the major Saccharomyces cerevisiae Cdk, Cdc28p. We found that the majority of Cdc28p was phosphorylated on its activating threonine (Thr-169) throughout the cell cycle. The extent of Thr-169 phosphorylation was similar for monomeric Cdc28p and Cdc28p bound to cyclin. By varying the order of the addition of cyclin and Cak1p, we determined that Cdc28p was activated most efficiently when it was phosphorylated before cyclin binding. Furthermore, we found that a Cdc28p(T169A) mutant, which cannot be phosphorylated, bound cyclin less well than wild-type Cdc28p in vivo. These results suggest that unphosphorylated Cdc28p may be unable to bind tightly to cyclin. We propose that Cdc28p is normally phosphorylated by Cak1p before it binds cyclin. This activation pathway contrasts with that in higher eukaryotes, in which cyclin binding appears to precede activating phosphorylation. PMID- 10793139 TI - Glycosylphosphatidylinositol biosynthesis defects in Gpi11p- and Gpi13p-deficient yeast suggest a branched pathway and implicate gpi13p in phosphoethanolamine transfer to the third mannose. AB - Glycosylphosphatidylinositols (GPIs) are critical for membrane anchoring and intracellular transport of certain secretory proteins. GPIs have a conserved trimannosyl core bearing a phosphoethanolamine (EthN-P) moiety on the third mannose (Man-3) through which the glycolipid is linked to protein, but diverse GPI precursors with EthN-Ps on Man-1 and Man-2 have also been described. We report on two essential yeast genes whose products are required late in GPI assembly. GPI11 (YDR302w) encodes a homologue of human Pig-Fp, a protein implicated in the addition of EthN-P to Man-3. PIG-F complements the gpi11 deletion, but the rescued haploids are temperature sensitive. Abolition of Gpi11p or Pig-Fp function in GPI11 disruptants blocks GPI anchoring and formation of complete GPI precursors and leads to accumulation of two GPIs whose glycan head groups contain four mannoses but differ in the positioning and number of side chains, probably EthN-Ps. The less polar GPI bears EthN-P on Man-2, whereas the more polar lipid has EthN-P on Man-3. The latter finding indicates that Gpi11p is not required for adding EthN-P to Man-3. Gpi13p (YLL031cp), a member of a family of phosphoryltransferases, is a candidate for the enzyme responsible for adding EthN-P to Man-3. Depletion of Gpi13p in a Gpi11p-defective strain prevents formation of the GPI bearing EthN-P on Man-3, and Gpi13p-deficient strains accumulate a Man(4)-GPI isoform that bears EthN-P on Man-1. We further show that the lipid accumulation phenotype of Gpi11p-deficient cells resembles that of cells lacking Gpi7p, a sequence homologue of Gpi13p known to add EthN-P to Man-2 of a late-stage GPI precursor. This result suggests that in yeast a Gpi11p deficiency can affect EthN-P addition to Man-2 by Gpi7p, in contrast to the Pig Fp defect in mammalian cells, which prevents EthN-P addition to Man-3. Because Gpi11p and Pig-Fp affect EthN-P transfer to Man-2 and Man-3, respectively, these proteins may act in partnership with the GPI-EthN-P transferases, although their involvement in a given EthN-P transfer reaction varies between species. Possible roles for Gpi11p in the supply of the EthN-P donor are discussed. Because Gpi11p- and Gpi13p-deficient cells accumulate isoforms of Man(4)-GPIs with EthN-P on Man 2 and on Man-1, respectively, and because the GPIs that accumulate in Gpi11p defective strains are likely to have been generated independently of one another, we propose that the yeast GPI assembly pathway is branched. PMID- 10793140 TI - gdt1, a new signal transduction component for negative regulation of the growth differentiation transition in Dictyostelium discoideum. AB - Discoidin I expression was used as a marker to screen for mutants affected in the growth-differentiation transition (GDT) of Dictyostelium. By REMI mutagenesis we have isolated mutant 2-9, an overexpressor of discoidin I. It displays normal morphogenesis but shows premature entry into the developmental cycle. The disrupted gene was denominated gdt1. The mutant phenotype was reconstructed by disruptions in different parts of the gene, suggesting that all had a complete loss of function. gdt1 was expressed in growing cells; the levels of protein and mRNA appear to increase with cell density and rapidly decrease with the onset of development. gdt1 encodes a 175-kDa protein with four putative transmembrane domains. In the C terminus, the derived amino acid sequence displays some similarity to the catalytic domain of protein kinases. Mixing experiments demonstrate that the gdt1(-) phenotype is cell autonomous. Prestarvation factor is secreted at wild-type levels. The response to folate, a negative regulator of discoidin expression, was not impaired in gdt1 mutants. Cells that lack the G protein alpha2 display a loss of discoidin expression and do not aggregate. gdt1( )/Galpha2(-) double mutants show no aggregation but strong discoidin expression. This suggests that gdt1 is a negative regulator of the GDT downstream of or in a parallel pathway to Galpha2. PMID- 10793142 TI - Endoplasmic reticulum quality control of oligomeric membrane proteins: topogenic determinants involved in the degradation of the unassembled Na,K-ATPase alpha subunit and in its stabilization by beta subunit assembly. AB - The molecular nature of determinants that mediate degradation of unassembled, polytopic subunits of oligomeric membrane proteins and their stabilization after partner subunit assembly is largely unknown. Expressing truncated Na,K-ATPase alpha subunits alone or together with beta subunits, we find that in unassembled alpha subunits neither the four N-terminal transmembrane segments acting as efficient alternating signal anchor-stop transfer sequences nor the large, central cytoplasmic loop exposes any degradation signal, whereas poor membrane insertion efficiency of C-terminal membrane domains M5, M7, and M9 coincides with the transient exposure of degradation signals to the cytoplasmic side. beta assembly with an alpha domain comprising at least D902 up to Y910 in the extracytoplasmic M7/M8 loop is necessary to stabilize Na,K-ATPase alpha subunits by favoring M7/M8 membrane pair formation and by protecting a degradation signal recognized from the endoplasmic reticulum (ER) lumenal side. Thus our results suggest that ER degradation of Na,K-ATPase alpha subunits is 1) mainly mediated by folding defects caused by inefficient membrane insertion of certain membrane domains, 2) a multistep process, which involves proteolytic and/or chaperone components acting from the ER lumenal side in addition to cytosolic, proteasome related factors, and 3) prevented by partner subunit assembly because of direct protection and retrieval of degradation signals from the cytoplasm to the ER lumenal side. These results likely represent a paradigm for the ER quality control of unassembled, polytopic subunits of oligomeric membrane proteins. PMID- 10793141 TI - High-resolution FRET microscopy of cholera toxin B-subunit and GPI-anchored proteins in cell plasma membranes. AB - "Lipid rafts" enriched in glycosphingolipids (GSL), GPI-anchored proteins, and cholesterol have been proposed as functional microdomains in cell membranes. However, evidence supporting their existence has been indirect and controversial. In the past year, two studies used fluorescence resonance energy transfer (FRET) microscopy to probe for the presence of lipid rafts; rafts here would be defined as membrane domains containing clustered GPI-anchored proteins at the cell surface. The results of these studies, each based on a single protein, gave conflicting views of rafts. To address the source of this discrepancy, we have now used FRET to study three different GPI-anchored proteins and a GSL endogenous to several different cell types. FRET was detected between molecules of the GSL GM1 labeled with cholera toxin B-subunit and between antibody-labeled GPI anchored proteins, showing these raft markers are in submicrometer proximity in the plasma membrane. However, in most cases FRET correlated with the surface density of the lipid raft marker, a result inconsistent with significant clustering in microdomains. We conclude that in the plasma membrane, lipid rafts either exist only as transiently stabilized structures or, if stable, comprise at most a minor fraction of the cell surface. PMID- 10793143 TI - Identification and characterization of Saccharomyces cerevisiae Cdc6 DNA-binding properties. AB - Recent studies have shown that Cdc6 is an essential regulator in the formation of DNA replication complexes. However, the biochemical nature of the Cdc6 molecule is still largely unknown. In this report, we present evidence that the Saccharomyces cerevisiae Cdc6 protein is a double-stranded DNA-binding protein. First, we have demonstrated that the purified yeast Cdc6 can bind to double stranded DNA (dissociation constant approximately 1 x 10(-7) M), not to single stranded DNA, and that the Cdc6 molecule is a homodimer in its native form. Second, we show that GST-Cdc6 fusion proteins expressed in Escherichia coli bind DNA in an electrophoretic mobility shift assay. Cdc6 antibodies and GST antibodies, but not preimmune serum, induce supershifts of GST-Cdc6 and DNA complexes in these assays, which also showed that GST-Cdc6 binds to various DNA probes without apparent sequence specificity. Third, the minimal requirement for the binding of Cdc6 to DNA has been mapped within its N-terminal 47-amino acid sequence (the NP6 region). This minimal binding domain shows identical DNA binding properties to those possessed by full-length Cdc6. Fourth, the GST-NP6 protein competes for DNA binding with distamycin A, an antibiotic that chelates DNA within the minor groove of the A+T-rich region. Finally, site-direct mutagenesis studies revealed that the (29)KRKK region of Cdc6 is essential for Cdc6 DNA-binding activity. To further elucidate the function of Cdc6 DNA binding in vivo, we demonstrated that a binding mutant of Cdc6 fails to complement either cdc6-1 temperature-sensitive mutant cells or Deltacdc6 null mutant cells at the nonpermissive temperature. The mutant gene also conferred growth impairments and increased the plasmid loss in its host, indicative of defects in DNA synthesis. Because the mutant defective in DNA binding also fails to stimulate Abf1 ARS1 DNA binding activity, our results suggest that Cdc6 DNA-binding activity may play a pivotal role in the initiation of DNA replication. PMID- 10793144 TI - Truncation mutants of the tight junction protein ZO-1 disrupt corneal epithelial cell morphology. AB - The tight junction is the most apical intercellular junction of epithelial cells and regulates transepithelial permeability through the paracellular pathway. To examine possible functions for the tight junction-associated protein ZO-1, C terminally truncated mutants and a deletion mutant of ZO-1 were epitope tagged and stably expressed in corneal epithelial cell lines. Only full-length ZO-1 and one N-terminal truncation mutant targeted to cell borders; other mutants showed variable cytoplasmic distributions. None of the mutants initially disrupted the localization of endogenous ZO-1. However, long-term stable expression of two of the N-terminal mutants resulted in a dramatic change in cell shape and patterns of gene expression. An elongated fibroblast-like shape replaced characteristic epithelial cobblestone morphology. In addition, vimentin and smooth muscle actin expression were up-regulated, although variable cytokeratin expression remained, suggesting a partial transformation to a mesenchymal cell type. Concomitant with the morphological change, the expression of the integral membrane tight junction protein occludin was significantly down-regulated. The localizations of endogenous ZO-1 and another family member, ZO-2, were disrupted. These findings suggest that ZO-1 may participate in regulation of cellular differentiation. PMID- 10793145 TI - HRD gene dependence of endoplasmic reticulum-associated degradation. AB - Work from several laboratories has indicated that many different proteins are subject to endoplasmic reticulum (ER) degradation by a common ER-associated machinery. This machinery includes ER membrane proteins Hrd1p/Der3p and Hrd3p and the ER-associated ubiquitin-conjugating enzymes Ubc7p and Ubc6p. The wide variety of substrates for this degradation pathway has led to the reasonable hypothesis that the HRD (Hmg CoA reductase degradation) gene-encoded proteins are generally involved in ER protein degradation in eukaryotes. We have tested this model by directly comparing the HRD dependency of the ER-associated degradation for various ER membrane proteins. Our data indicated that the role of HRD genes in protein degradation, even in this highly defined subset of proteins, can vary from absolute dependence to complete independence. Thus, ER-associated degradation can occur by mechanisms that do not involve Hrd1p or Hrd3p, despite their apparently broad envelope of substrates. These data favor models in which the HRD gene-encoded proteins function as specificity factors, such as ubiquitin ligases, rather than as factors involved in common aspects of ER degradation. PMID- 10793146 TI - Activation of cdc42, rac, PAK, and rho-kinase in response to hepatocyte growth factor differentially regulates epithelial cell colony spreading and dissociation. AB - Hepatocyte growth factor (HGF), the ligand for the Met receptor tyrosine kinase, is a potent modulator of epithelial-mesenchymal transition and dispersal of epithelial cells, processes that play crucial roles in tumor development, invasion, and metastasis. Little is known about the Met-dependent proximal signals that regulate these events. We show that HGF stimulation of epithelial cells leads to activation of the Rho GTPases, Cdc42 and Rac, concomitant with the formation of filopodia and lamellipodia. Notably, HGF-dependent activation of Rac but not Cdc42 is dependent on phosphatidylinositol 3-kinase. Moreover, HGF induced lamellipodia formation and cell spreading require phosphatidylinositol 3 kinase and are inhibited by dominant negative Cdc42 or Rac. HGF induces activation of the Cdc42/Rac-regulated p21-activated kinase (PAK) and c-Jun N terminal kinase, and translocation of Rac, PAK, and Rho-dependent Rho-kinase to membrane ruffles. Use of dominant negative and activated mutants reveals an essential role for PAK but not Rho-kinase in HGF-induced epithelial cell spreading, whereas Rho-kinase activity is required for the formation of focal adhesions and stress fibers in response to HGF. We conclude that PAK and Rho kinase play opposing roles in epithelial-mesenchymal transition induced by HGF, and provide new insight regarding the role of Cdc42 in these events. PMID- 10793147 TI - Role of actin and Myo2p in polarized secretion and growth of Saccharomyces cerevisiae. AB - We examined the role of the actin cytoskeleton in secretion in Saccharomyces cerevisiae with the use of several quantitative assays, including time-lapse video microscopy of cell surface growth in individual living cells. In latrunculin, which depolymerizes filamentous actin, cell surface growth was completely depolarized but still occurred, albeit at a reduced level. Thus, filamentous actin is necessary for polarized secretion but not for secretion per se. Consistent with this conclusion, latrunculin caused vesicles to accumulate at random positions throughout the cell. Cortical actin patches cluster at locations that correlate with sites of polarized secretion. However, we found that actin patch polarization is not necessary for polarized secretion because a mutant, bee1Delta(las17Delta), which completely lacks actin patch polarization, displayed polarized growth. In contrast, a mutant lacking actin cables, tpm1-2 tpm2Delta, had a severe defect in polarized growth. The yeast class V myosin Myo2p is hypothesized to mediate polarized secretion. A mutation in the motor domain of Myo2p, myo2-66, caused growth to be depolarized but with only a partial decrease in the level of overall growth. This effect is similar to that of latrunculin, suggesting that Myo2p interacts with filamentous actin. However, inhibition of Myo2p function by expression of its tail domain completely abolished growth. PMID- 10793148 TI - Complex regulation of the yeast heat shock transcription factor. AB - The yeast heat shock transcription factor (HSF) is regulated by posttranslational modification. Heat and superoxide can induce the conformational change associated with the heat shock response. Interaction between HSF and the chaperone hsp70 is also thought to play a role in HSF regulation. Here, we show that the Ssb1/2p member of the hsp70 family can form a stable, ATP-sensitive complex with HSF-a surprising finding because Ssb1/2p is not induced by heat shock. Phosphorylation and the assembly of HSF into larger, ATP-sensitive complexes both occur when HSF activity decreases, whether during adaptation to a raised temperature or during growth at low glucose concentrations. These larger HSF complexes also form during recovery from heat shock. However, if HSF is assembled into ATP-sensitive complexes (during growth at a low glucose concentration), heat shock does not stimulate the dissociation of the complexes. Nor does induction of the conformational change induce their dissociation. Modulation of the in vivo concentrations of the SSA and SSB proteins by deletion or overexpression affects HSF activity in a manner that is consistent with these findings and suggests the model that the SSA and SSB proteins perform distinct roles in the regulation of HSF activity. PMID- 10793149 TI - The yeast heat shock transcription factor changes conformation in response to superoxide and temperature. AB - In vitro DNA-binding assays demonstrate that the heat shock transcription factor (HSF) from the yeast Saccharomyces cerevisiae can adopt an altered conformation when stressed. This conformation, reflected in a change in electrophoretic mobility, requires that two HSF trimers be bound to DNA. Single trimers do not show this change, which appears to represent an alteration in the cooperative interactions between trimers. HSF isolated from stressed cells displays a higher propensity to adopt this altered conformation. Purified HSF can be stimulated in vitro to undergo the conformational change by elevating the temperature or by exposing HSF to superoxide anion. Mutational analysis maps a region critical for this conformational change to the flexible loop between the minimal DNA-binding domain and the flexible linker that joins the DNA-binding domain to the trimerization domain. The significance of these findings is discussed in the context of the induction of the heat shock response by ischemic stroke, hypoxia, and recovery from anoxia, all known to stimulate the production of superoxide. PMID- 10793150 TI - Mammalian spindle orientation and position respond to changes in cell shape in a dynein-dependent fashion. AB - In animal cells, positioning of the mitotic spindle is crucial for defining the plane of cytokinesis and the size ratio of daughter cells. We have characterized this phenomenon in a rat epithelial cell line using microscopy, micromanipulation, and microinjection. Unmanipulated cells position the mitotic spindle near their geometric center, with the spindle axis lying roughly parallel to the long axis of the cell. Spindles that were initially misoriented underwent directed rotation and caused a delay in anaphase onset. To gain further insight into this process, we gently deformed cells with a blunted glass needle to change the spatial relationship between the cortex and spindle. This manipulation induced spindle movement or rotation in metaphase and/or anaphase, until the spindle reached a proper position relative to the deformed shape. Spindle positioning was inhibited by either treatment with low doses of nocodazole or microinjection of antibodies against dynein, apparently due to the disruption of the organization of dynein and/or astral microtubules. Our results suggest that mitotic cells continuously monitor and maintain the position of the spindle relative to the cortex. This process is likely driven by interactions among astral microtubules, the motor protein dynein, and the cell cortex and may constitute part of a mitotic checkpoint mechanism. PMID- 10793151 TI - The p21 Rho-activating toxin cytotoxic necrotizing factor 1 is endocytosed by a clathrin-independent mechanism and enters the cytosol by an acidic-dependent membrane translocation step. AB - Cytotoxic necrotizing factor 1 (CNF1), a protein produced by pathogenic strains of Escherichia coli, activates the p21 Rho-GTP-binding protein, inducing a profound reorganization of the actin cytoskeleton. CNF1 binds to its cell surface receptor on HEp-2 cells with high affinity (K(d) = 20 pM). In HEp-2 cells the action of CNF1 is not blocked in the presence of filipin, a drug described to reduce cholera toxin internalization by the caveolae-like mechanism. Moreover, HEp-2 cells, which express a dominant negative form of proteins that impair the formation of clathrin coated-vesicles and internalization of transferrin (Eps15, dynamin or intersectin-Src homology 3), are still sensitive to CNF1. In this respect, the endocytosis of CNF1 is similar to the plant toxin ricin. However, unlike ricin toxin, CNF1 does not cross the Golgi apparatus and requires an acidic cell compartment to transfer its enzymatic activity into the cytosol in a manner similar to that required by diphtheria toxin. As shown for diphtheria toxin, the pH-dependent membrane translocation step of CNF1 could be mimicked at the level of the plasma membrane by a brief exposure to a pH of thromboxane A(2) thromboxane A(2) receptor. PMID- 10793266 TI - Pharmacological effects of novel quinone compounds, 6-(fluorinated-phenyl)amino 5,8-quinolinediones, on inhibition of drug-induced relaxation of rat aorta and their putative action mechanism. AB - Two 6-(fluorinated-phenyl)amino-5,8-quinolinedione derivatives, OQ21 and OQ1, were newly synthesized as potent inhibitors of endothelial-dependent vasorelaxation. The purpose of the present study was to investigate the effects of OQ21 and OQ1 on different types of vasorelaxation and to pursue their action mechanisms. For acetylcholine both compounds, at a low concentration (0.1 microM), reduced the maximal response with increase of EC(50) values. OQ21 is a novel quinone compound and showed a more potent and efficacious inhibitory effect on acetylcholine-induced relaxation of rat aorta than that of LY83583 (6-anilino 5,8-quinolinedione). Relatively high concentrations (1 microM) of OQ21 and OQ1 inhibited the sodium nitroprusside-induced relaxation of endothelium-denuded ring, producing rightward shifts of the curve for sodium nitroprusside without altering the maximal response. They also prevented acetylcholine and sodium nitroprusside-induced elevations of cyclic GMP. In addition, OQ21 and OQ1 (1 microM) significantly decreased (52-72%) the sensitivity of L-arginine-induced relaxation of precontracted endothelium-denuded aortic rings from lipopolysaccaride-treated (20 mg/kg, i.p.) rats. The inhibitory effect of OQ21 on endothelium-dependent vasodilation was enhanced by N(G)-nitro-L-arginine, which inhibits nitric oxide synthase (NOS) by binding the oxygenase domain of the enzyme, but not by diphenylendiodonium, which inhibits NOS by binding to the reductase domain of the enzyme. Treatment of blood vessels with OQ21 or OQ1 showed a significant increase in chemiluminescence output, which was prevented by adding superoxide dismutase, suggesting that superoxide generation is involved in the action mechanism for OQ21. Present results indicate that a novel naphthoquinone compound, OQ21, potently inhibits endothelial NOS, possibly by interacting with the reductase domain of the enzyme, which leads to induce superoxide formation. The new benzoquinone compounds, OQ21 and OQ1, inhibit not only endothelium-dependent vasorelaxation but also endothelium-independent relaxation induced by exogenous NO generated from a nitrovasodilator via the reduction of cyclic GMP. They also reduced L-arginine-induced vasorelaxation in endotoxin-treated rats, indicating their possession of inhibitory effect on inducible NOS. PMID- 10793267 TI - Renal electrolyte and fluid handling in the rat following chloroquine and/or ethanol administration. AB - We postulated that chloroquine and/or ethanol affect plasma arginine vasopressin (AVP) concentrations to alter renal function. Therefore, we studied the effects of chloroquine and/or ethanol on plasma AVP concentrations and fluid, urinary Na(+) and K(+) outputs in separate groups of anaesthetized Sprague-Dawley (SD) rats challenged with a continuous jugular infusion of 0.077 M NaCl at 150 microl.min(-1). After a 3-h equilibration period, vehicle, chloroquine (0.06 microg. min(-1)), ethanol (2.4 or 24 microg.min(-1)) or both chloroquine and ethanol were added to the infusate after 1 h (control) for 1 h 20 min (treatment). The animals were switched back to the infusate alone for the final 1 h 40 min recovery periods. Urine flow Na(+) and K(+) excretion rates were determined at 20-min intervals over the subsequent 4-h postequilibration period. Blood was collected from separate groups of animals at the end of treatment period or equivalent time for control animals for measurement of plasma aldosterone and AVP concentrations by radioimmunoassay. Simultaneous chloroquine and ethanol infusion significantly (p < 0.01) increased plasma chloroquine concentrations in an ethanol dose-dependent manner by comparison with animals administered chloroquine alone. Chloroquine infusion alone (0.06 microg.min(-1)) and/or ethanol (2.4 or 24 microg.min(-1)) elevated plasma AVP concentrations from 9.73 +/- 1.64 fmol.l(-1) in control rats to 15.65 +/- 2.49 fmol.l(-1), 17. 39 +/- 4.21 fmol.l(-1), and 33.87 +/- 6.18 fmol.l(-1), respectively. Separate administration of chloroquine or ethanol at low dose rates increased urinary Na(+) excretion rates. We conclude that the impairment of renal electrolyte handling associated with chloroquine administration may be exacerbated by ethanol. PMID- 10793268 TI - The effects of growth hormone-releasing peptide-2 (GHRP-2) on the release of growth hormone and growth performance in swine. AB - The effects of GHRP-2 (also named KP102), a new growth hormone-releasing peptide, on the release of growth hormone (GH) and growth performance were examined in swine. The single intravenous (i. v.) injection of GHRP-2 at doses of 2, 10, 30 and 100 microg/kg body weight (BW) to cross-bred castrated male swine stimulated GH release in a dose-dependent manner, with a return to the baseline by 120 min. The peak GH concentrations and GH areas under the response curves (GH AUCs) for 180 min after the injections of GHRP-2 were higher (P < 0.05) than those after the injection of saline. The GH responses to repeated i.v. injections of GHRP-2 (30 microg/kg BW) at 2-h intervals for 6 h were decreased after each injection. The chronic subcutaneous (s.c.) administration of GHRP-2 (30 microg/kg BW) once daily for 30 days consistently stimulated GH release. The GH AUCs for 300 min after the injections on d 1, 10 and 30 of treatment in GHRP-2-treated swine were higher than those in saline-treated swine. However, chronic administration of GHRP-2 caused a partial attenuation of GH response between d 1 and 10 of treatment. The chronic s.c. administration of GHRP-2 also increased average daily gain for the entire treatment period by 22.35% (P < 0.05) and feed efficiency (feed/gain) by 20.64% (P < 0.01) over the saline control values, but did not significantly affect daily feed intake. These results indicate that GHRP-2 stimulates GH release and enhancing growth performance in swine. PMID- 10793269 TI - The effect of growth hormone-releasing peptide-2 (KP102) administration on plasma insulin-like growth factor (IGF)-1 and IGF-binding proteins in Holstein steers on different planes of nutrition. AB - This study was conducted to investigate the nutrition-dependent changes in insulin-like growth factor (IGF)-1 and IGF-binding proteins (IGFBPs) with growth hormone releasing peptide-2 (D-Ala-D-betaNal-Ala-Trp-D-Phe-Lys-NH(2); GHRP-2 or KP102) treatment in growing Holstein steers. Eight 13 month-old Holstein steers were grouped on two levels of feed intake (high intake (HI); 2.43% body weight or low intake (LI); 1.22%) and each group was daily injected with KP102 (12.5 microg/kg body weight/day) or saline solution into the jugular vein during 6-day period. The concentration of plasma GH showed an increase after an i.v. bolus injection of KP102 on Day 1 and Day 6 in both the LI and HI groups. Plasma IGF-1 began to increase 10 hr following an i.v. bolus injection of KP102, but this was only observed in the HI group (P < 0.05). Also, the plasma IGF-1 in the HI group with daily injections was significantly greater than the LI group from Day 1 of KP102 administration (P < 0.05). It reached maximum values of 125.1 +/- 7.6 ng/ml after Day 2, and returned to pre-injection levels after Day 4, however, no change in plasma IGF-1 was observed in LI with administration of KP102. During 6 days of treatment, plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 were significantly higher in KP102 treated steers but only in the HI group (P < 0.05). Plasma 34 kDa IGFBP 2 decreased in the HI group and did not show any change following an injection of KP102. In conclusion, the effect of stimulated endogenous GH with KP102 administration increased plasma IGF-1, 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 levels in the HI group of growing Holstein steers, but not in the LI one. Thus, we strongly believe that the plasma IGF-1 and IGFBPs response to KP102 treatment is modulated by the nutritional status of growing Holstein steers and the increased plasma IGF-1 concentration with KP102 treatment may be regulated by plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 in Holstein steers. PMID- 10793271 TI - A chicken leptin-specific radioimmunoassay. AB - Recombinant chicken leptin was used to produce an antiserum in order to develop a specific and sensitive radioimmunoassay (RIA) for chicken leptin in plasma and serum. We have used either murine or chicken leptin as tracer and competition curves were performed using recombinant chicken leptin. Variations in leptin plasma levels in different chicken strains and various nutritional states were correlated with the physiological status. Leptin plasma concentrations were regulated by the nutritional state with higher levels in the fed state as compared to the fasted state (3.36 +/- 0. 13 versus 2.78 +/- 0.11 ng/ml) and being dependent upon the age. Higher leptin levels were found in 22 week-old as compared to 15 week-old layer chickens (2.709 +/- 0.172 versus 1.478 +/- 0.102 ng/ml). We have also shown that the multispecies leptin RIA kit (LINCO Inc.) underestimated leptinemia compared to the chicken leptin- specific RIA reported here. In conclusion the RIA developed in the present study is specific to the chicken and thus may be considered as powerful tool for investigating the physiological significance of leptin in chickens. PMID- 10793270 TI - Transcriptional regulation of pituitary synthesis and secretion of growth hormone in growing wethers and the influence of zeranol on these mechanisms. AB - This experiment evaluated relationships between pituitary messenger RNA levels of the transcription factor Pit-1, the growth hormone releasing-hormone receptor (GHRHR), and synthesis and secretion of GH in growing wethers. The experiment also evaluated the influence of the estrogenic compound, zeranol, on these relationships. Seventy wethers that were 9.5 +/- 1 day of age were randomly assigned to a control group or to one of three zeranol treatment groups that were implanted (12 mg, Ralgro) at 0, 45, and (or) 90 days of age. Twenty-eight days after implantation (i.e., Days 28, 73, 118) and on Day 135, sera were collected serially from wethers (n > or = 5) from each group and then their pituitary was collected. As wethers gained weight with age, the pituitary increased in size and so did the relative message levels of Pit-1 and GH (effect of time, P < 0.01). However, as wethers reached 135 days of age, serum concentrations of GH had declined while concentrations of IGF-I had increased (linear contrast, P < 0.01). Additionally, zeranol increased serum concentrations of GH and IGF-I and this effect on GH appeared to be a consequence of increased pulse amplitude, particularly at 73 and 118 days of age (treatment x time, P 0.05). During follow-up, 3 patients who underwent CND type 3 (1.6%) and 12 who underwent RND (4.1%) had regional recurrences. The difference between recurrence rates was not statistically significant (P > 0. 05). A conclusion was reached that CND type 3 safely provided regional cancer control in N0 laryngeal cancer and that it might be performed to decrease the morbidity of RND. PMID- 10793357 TI - Surgical treatment of labyrinthine fistula caused by cholesteatoma. AB - In a 144-month period, 27 cases of labyrinthine fistula (LF) were seen, and 360 mastoid operations were performed; the LF prevalence was 7.5%. Primary symptoms were hypoacusis, otorrhea, vertigo, tinnitus, and otalgia. All patients underwent preoperative CT scans and preoperative audiometry. LF diagnosis was made before surgery for 93% of patients on the basis of symptoms, signs, and imaging studies. With respect to surgical technique, the canal-wall-down procedure was performed in 92%, and the canal-wall-up procedure was performed in 8%. In 88% of patients the fistula was located in the horizontal semicircular canal. In 96% of patients the cholesteatoma matrix was removed, and the fistula was sealed; in 4% of patients the matrix was left. With a follow-up of 13 years, vertigo disappeared in 96% of patients, and hearing remained unchanged in 70% of patients. Further complications of chronic otitis media existed in approximately half of the patients with LF. Open surgery with removal of the cholesteatoma matrix and sealing of the fistula with temporalis fascia in a canal-wall-down manner is a safe procedure that can make vertigo disappear and helps to preserve cochlear function. PMID- 10793358 TI - Acoustic analysis of the voice in phonatory fistuloplasty after total laryngectomy. AB - A comparative study of the voice with sustained phonation of the vowel /a/ was made in 3 groups of male patients: (1) 20 patients receiving total laryngectomy for epidermoid carcinoma of the larynx who had acquired good voice quality after a phonatory fistuloplasty with a Herrmann voice prosthesis; (2) 20 patients undergoing total laryngectomy for epidermoid carcinoma of the larynx who had learned esophageal speech; and (3) 20 subjects with normal voices. Statistical analysis yielded significant differences in fundamental voice frequency between the 3 groups, with the patients with phonatory prostheses revealing the closest to a normal voice. For other parameters used, such as jitter, shimmer, and harmonics/noise ratio, voice quality with a phonatory prosthesis was similar to that obtained with esophageal speech. PMID- 10793359 TI - Hearing results in the second stage of open mastoidectomy: A comparison of the different techniques. AB - OBJECTIVE: The goal of this study was to compare the hearing results obtained through different hearing-restoration techniques in open-cavity mastoidectomy. METHODS AND MATERIAL: A total of 116 ears operated on during the second stage of open mastoidectomy were studied. The hearing gain was registered as the difference between preoperative and 1-year postoperative conversational frequency means (500, 1000, 2000 Hz). High-pitched frequencies (4000 Hz) were used equally. Two groups of studies were carried out: in one study the stapes was intact, and in the other the stapes arch was absent. The columella techniques we examined, from a hearing point of view, are total and partial ossicular replacement prostheses, ossiculoplasty, cartilage with or without perichondrium, and the myringostapediopexy. RESULTS AND CONCLUSIONS: In the group of patients in whom the stapes was intact, the differences in high-pitched frequencies found between the different techniques were negligible, but the differences were quite significant in conversational frequencies, where the partial ossicular replacement prosthesis turned out to be the technique with the worst results. In ears in which only the footplate was present, the total ossicular replacement prosthesis gave better results than any other technique, both in conversational frequencies and in 4000-Hz frequency. PMID- 10793361 TI - Skin prick test reactivity to foods in adult Malaysians with rhinitis. AB - The aim of the study was to determine the incidence of food and house dust mite (HDM) allergy in patients with nasal congestion and rhinorrhea attending the Otorhinolaryngology Clinic, National University of Malaysia, Kuala Lumpur. This was a prospective matched, controlled study of patients skin prick tested with commercial food and common aeroallergens. The participants were 148 Malaysian adults with symptoms of nasal congestion and rhinorrhea and 113 adult Malaysian control subjects without rhinitis symptoms. The skin prick test (SPT) was used to evaluate 11 foods common to the Malaysian diet and 3 HDM inhalants. Forty-eight percent of the patients with rhinitis had positive SPT results to foods, compared with 4.4% of control subjects (P < 0.05). The most commonly implicated foods were shrimp (48%) and rice (30%), which are common in the Malaysian diet. Seventy-two percent of rhinitis patients had positive SPT results to HDM, compared with 22.2% of control subjects (P < 0.05). Patients with rhinitis also had significantly more gastrointestinal problems than control subjects (P < 0.05). The incidences of HDM and food allergy are significantly greater in Malaysian adults with rhinitis symptoms than in control subjects without rhinitis. The effect of avoidance or immunotherapy awaits further study. PMID- 10793360 TI - Complications of postoperative radiation therapy after partial laryngectomy in supraglottic cancer: A long-term evaluation. AB - This retrospective study, based on a series of 90 patients with invasive squamous cell carcinoma of the supraglottis, was designed to document the functional outcome and complications after postoperative radiation therapy following partial laryngeal surgery. The surgical procedure was a standard supraglottic laryngectomy in 62 patients and a supracricoid partial laryngectomy in 28 patients. All of the patients had an unremarkable postoperative course and achieved locoregional control. The average dose delivered to the remaining larynx was 51.2 Gy (range 25-71 Gy). The average dose delivered to the neck was 50.6 Gy (range 22-70 Gy). The patients were treated at 180-cGy per fractions in a continuous course technique with a cobalt 60 beam. In 5 patients (5.5%) complications led to cessation of postoperative radiation therapy, and the total dose delivered to the remaining larynx and neck was less than 40 Gy. All patients were followed up for a minimum of 10 years or until death. The 5-, 10-, and 15 year actuarial survival estimates were 71. 5%, 44.3%, and 36.3%, respectively. The 5-, 10-, and 15-year actuarial severe complication estimates were all 11.2%. Overall, severe complications occurred in 15 patients. Severe complications led to death in 3 patients (3.3%), permanent gastrostomy in 3 (3.3%), and permanent tracheostomy in 1 (1.1%). A severe complication never resulted in completion of total laryngectomy. In univariate analysis, the mean dose delivered to the larynx was the only variable statistically related to the incidence of a severe complication. The mean dose delivered to the larynx was statistically higher (P = 0.014) in patients who had severe complications (60 Gy) than in patients who did not (50 Gy). PMID- 10793362 TI - Therapeutic dilemmas in the management of thyroid cancer with laryngotracheal involvement. AB - Invasion of the larynx and trachea by thyroid cancer is an uncommon but difficult problem. There is no consensus on indication for or extent of surgery, particularly when there is a requirement for airway reconstruction. From 1989 through 1996, we treated 22 patients with thyroid carcinoma with invasion of the larynx and trachea. Seventeen of these patients had recurrent disease. We applied radioactive iodine therapy after regional ablative surgery to resectable tumors with or without lung metastasis, larynx-preserving surgery to extraluminal or small intraluminal tumors restricted to the short segment of trachea, or total laryngectomy to recurrent tumors deeply invading the cartilage framework of the larynx. We performed arytenoid adduction or thyroplasty in one stage if the recurrent laryngeal nerve was paralyzed or resected intraoperatively. We could get relatively good survival and functional results by aggressive surgical treatment in 20 patients, but the disease was inoperable in 2 patients. It is stressed that head and neck surgeons who have to deal with cancer of the thyroid should not only be familiar with various techniques of airway reconstruction and voice rehabilitation but also must be aware of the biologic behavior of the thyroid carcinoma. PMID- 10793363 TI - Tracheostomy tube obstruction caused by an overinflated cuff. PMID- 10793364 TI - Facial vein thrombophlebitis: A rare but potentially lethal entity. PMID- 10793365 TI - Endoscopic repair of anterior cranial fossa dural defect in an unusual case of frontal sinus trauma. PMID- 10793366 TI - Schwannoma of the larynx presenting with difficult swallowing. PMID- 10793367 TI - Cemento-ossifying fibroma of the upper gingivae. PMID- 10793368 TI - Maxillary sinus mass: the case for the unerupted third molar. PMID- 10793369 TI - Human aural myiasis. PMID- 10793370 TI - Airway obstruction caused by bee sting macroglossia. PMID- 10793371 TI - Experimental topical ototoxicity investigations: can we underestimate unilateral ototoxicity because of cross-hearing? PMID- 10793372 TI - Experimental topical ototoxicity investigations: can we underestimate unilateral ototoxicity because of cross-hearing? PMID- 10793373 TI - Asymmetric sensorineural hearing loss. PMID- 10793374 TI - The association of prosthodontists of canada-founded in 1973 PMID- 10793375 TI - The american academy of esthetic dentistry-founded in 1975 PMID- 10793376 TI - Evidence-based dentistry: prognosis. PMID- 10793377 TI - Bleaching and temporomandibular disorder using a half tray design: a clinical report. AB - A maxillary soft, custom-fitted tray was fabricated for a patient to perform nightguard vital bleaching. Treatment was interrupted after the patient experienced pain in the temporomandibular joint area shortly after wearing the bleaching prosthesis. The tray was trimmed so the labial, incisal and buccal cusps were covered and Sc) the patient had complete tooth-to-tooth contact in the maximum intercuspal position. The prosthesis proved to be retentive even without the presence of the bleaching material. The thick, sticky bleaching material was contained in the half tray design and the tray was held in place. The patient was able to continue the bleaching process for the 2-week duration necessary to achieve successful lightening of the teeth without further TMD symptoms. PMID- 10793379 TI - Prevention of tongue biting with a removable oral device: a clinical report. AB - A method of preventing tongue biting with a removal device has been described in this clinical report. Restraint of the tongue may be necessary to promote healing by preventing repetitive tongue biting or as a preventive measure after a surgery or an injury. Any device that is used to restrain the tongue should be removable to avoid prolonged interference with oral hygiene, swallowing, mastication, and speech. A removable device is desirable for long-term use by comatose or semicomatose patients. PMID- 10793378 TI - Etched porcelain veneer restoration of a primary tooth: a clinical report. AB - The treatment of a retained primary tooth in the adult dentition has always been a difficult clinical problem. In the anterior region of the mouth, the problem is further complicated by esthetic requirements. In this clinical report, a primary canine was recontoured both incisally and interproximally with an etched porcelain veneer to provide an esthetic result. The use of etched porcelain as an indirect veneer material, which has demonstrated clinical success, is an alternative to the requirements of conservative dentistry, namely, preservation of tooth substance and safeguarding primary tooth vitality. The procedure used in this clinical report differs from traditional procedures. The preparation was extended to the lingual surface of the tooth, which allowed for an increase of the incisal length and added a sufficient amount of porcelain. The etching procedure was performed twice by applying 30% phosphoric acid etchant gel to the prepared tooth. PMID- 10793380 TI - Metal reinforcement for implant-supported mandibular overdentures. AB - Complete and implant-supported mandibular overdenture bases can be a problem because they are especially prone to fracture. Metal reinforcement of the denture base can substantially reduce the incidence of fracture. This article describes 2 methods of reinforcing mandibular overdentures. The advantages of these methods and their indications are discussed. PMID- 10793381 TI - Controlled polymerization system for fabricating precise dentures. AB - STATEMENT OF PROBLEM: Most processed resin denture bases do not fit the cast accurately because of polymerization changes in the resin. The fit is more noticeable in the palate between the denture base and the cast. Therefore, development of a polymerization system that will improve the fit of a denture base to the cast is lack of worth considering to improve the fit and retention of the denture. PURPOSE: A newly developed polymerization system for dentures is presented that results in excellent adaptability of the denture base to the cast when compared with 2 other polymerization systems. MATERIAL AND METHODS: Forty five maxillary dentures were made with 3 resins and 3 polymerization systems (a new polymerization system, SR-Ivocap system, original microwave polymerization system). Each denture and cast was sectioned through the second molar area parallel to the posterior border. Discrepancies between denture base and stone cast were measured at several points immediately after polymerization and after 30 days of immersion in water. In addition, disks of 3 materials processed in each system were examined for absorption of water and hardness at 2 measurement stages. Data were analyzed with the split-plot or 2-way ANOVA test. RESULTS: Dentures made with the new polymerization system had significantly smaller discrepancies, compared with dentures processed with the SR-Ivocap system and the microwave method. The amount of water absorption of the resin disks processed by the new system was less than the disks made by the other 2 systems. Dentures made with the new system exhibited significantly better adaptation to the cast than those made with the other 2 systems. CONCLUSION: The new polymerization system produced a more accurate method of processing dentures when fit was compared with the fit of dentures made with the other 2 denture base resins. PMID- 10793382 TI - Relationship between the standards of removable partial denture construction, clinical acceptability, and patient satisfaction. AB - STATEMENT OF PROBLEM: Little is known about the importance of published fabrication standards in determining the outcome of treatment with mandibular distal extension removable partial dentures in patients of community practices. PURPOSE: This study describes mandibular partial dentures worn by patients from King County, Wash., and examines the validity of standards of design and fabrication by relating the standards to measures of clinical acceptability and patient satisfaction. MATERIAL AND METHODS: Eighty-two people treated in private dental practices who responded to a mail survey about satisfaction with a partial denture were examined. Eight standards of fabrication, overall clinical acceptability of the prosthesis, and tissue health were evaluated clinically. Patient satisfaction was assessed by questionnaire. Associations between variables were assessed by contingency tables and odds ratios. RESULTS: Half the prostheses met 4 or fewer of the 8 standards, and these were responsible for nearly all the displacement of tissue by the framework. Forty-three percent of the dentures (35/82) were rated clinically acceptable. Of the remaining dentures, 38 could be made acceptable by modifications. The remaining 9 dentures needed replacement. Sixty-three percent of the patients examined were satisfied with the dentures. There was a relationship between tissue health and the fabrication standards related to rest form, base extension, stress distribution, and framework fit. This study found no relation between tissue health and other design or fabrication features. None of the standards were found to be related to patient satisfaction. CONCLUSION: This study found partial support for the validity of design/fabrication standards for removable partial dentures. The most important standards are rest form, base extension, and stress distribution. The standards appear to be unrelated to patient satisfaction. PMID- 10793383 TI - Shade selection for resin-bonded fixed partial dentures. PMID- 10793384 TI - Long-term clinical results of galvano-ceramic and glass-ceramic individual crowns. AB - STATEMENT OF PROBLEM: Alternatives to metal-ceramic restorations should possess clinical durability before being recommended to the dentist. Longitudinal clinical studies are required for evaluation so innovative types of restorations can meet the expectations of dentists and patients. PURPOSE: This study compared the performance of galvano-ceramic restorations (Auvo Galvano Crown [ACG]) and glass-ceramic individual crowns (Dicor) based on longitudinal clinical trials. MATERIAL AND METHODS: A total of 769 galvano-ceramic single crowns (AGC) in 322 patients and 173 glass-ceramic individual crowns (Dicor) in 88 patients were reviewed for 8 years and a maximum of 11 years, respectively, after cementation. Risk of fracture was determined with use of a survival analysis (Kaplan and Meier). RESULTS: Partial ceramic cracking was observed at the time of the last recording of data in 11 galvano-ceramic crowns, 8 of the units remained in place. Two crowns became dislodged and 1 tooth exhibited a fractured root, despite an intact crown. One restoration was removed because of hypersensitivity and 1 with partial ceramic fracture. Forty-two of the glass-ceramic crowns were completely fractured. After a comparable 7 years under risk, 96.5% (+/-3.4; 95% confidence interval) of the galvano-ceramic premolar and molar crowns and 92% (+/- 8.5) of crowns placed on incisors and canines crowns were intact. The corresponding data for the glass-ceramic restorations were only 70% (+/- 10.6) in posterior and 82.7% (+/- 8. 1) in anterior quadrants. CONCLUSION: Long-term results of electroformed individual crown restorations were superior to glass-ceramic restorations. PMID- 10793385 TI - Marginal fit of metal ceramic restorations subjected to a standardized postsoldering technique. AB - The fit of a fixed partial denture (FPD) is fundamental for the clinical success of a restoration. When a postsoldering procedure is needed, a high-precision laboratory technique is necessary to not affect the fit of the FPD. This article evaluates whether a standardized postsoldering technique affected the marginal fit of a 3-unit high palladium alloy FPD. One hundred and eight measurements were made of 3-unit FPDs, fabricated in a high palladium alloy (2% Au-79% Pd-10% Cu-8% Ga) and constructed on 9 tin dies in vitro. After the castings were fabricated, specimens were measured at 3 specific points per abutment (distal, labial, lingual) before and after soldering. Differences were found in the marginal opening of both copings, before soldering (49.9 microm) and after soldering (48. 3 microm). There was no significant difference in the adaptation of the copings after the soldering procedure (P<.05). PMID- 10793386 TI - Influence of design and mode of loading on the fracture strength of all-ceramic resin-bonded fixed partial dentures: an in vitro study in a dual-axis chewing simulator. AB - STATEMENT OF PROBLEM: In a clinical study, all-ceramic resin-bonded fixed partial dentures showed a high rate of fractures within the first years of service but remained in function as cantilevered restorations. No data are available on the fracture strength of such cantilevered restorations. PURPOSE: This in vitro study evaluated the influence of design and mode of loading on the fracture strength of all-ceramic resin-bonded fixed partial dentures. MATERIAL AND METHODS: Forty eight frameworks were copy milled using the aluminum-oxide ceramic In-Ceram, glass-infiltrated, and circularly veneered with feldspathic porcelain to replace a maxillary incisor on a test cast. The airborne particle-abraded restorations were bonded to acid-etched human abutments using composite. Twenty-four restorations used a conventional 2-retainer design and another 24 restorations used a cantilevered single-retainer design. Subgroups of 8 specimens were subjected to a quasi-static load in the direction of the long axis of the abutments (0 degrees) or in an angle of 45 degrees. Additional subgroups were subjected to dynamic loading under 45 degrees with either 50 or 25 N in a dual axis chewing simulator until fracture. RESULTS: Mean fracture strengths, under 45 degree quasi-static loading, were between 134 and 174 N and under 0-degree loading about 233 N. Samples subjected to dynamic loading fractured after 25 to over 200,000 loading cycles. CONCLUSION: Direction of loading exhibited a significant influence on the fracture strength, regardless of the retainer design. The applied dynamic loading force, regardless of the retainer design, had a significant influence on the loading cycles until fracture. PMID- 10793387 TI - Clinical anatomy and palpability of the inferior lateral pterygoid muscle. AB - STATEMENT OF PROBLEM: The intraoral palpation technique of the inferior belly of the inferior lateral pterygoid (ILP) muscle is a standard diagnostic examination method for temporomandibular joint dysfunction syndrome, although different studies have revealed inconsistent results. PURPOSE: This study assessed the feasibility of the ILP muscle palpation by a simulated clinical setting. MATERIAL AND METHODS: Three dentists performed a bilateral palpation of the ILP muscle in 53 fresh and unfixed human cadavers and decided whether the muscle was palpable or unpalpable. In a second step, it was observed through the dissected infratemporal fossa, whether the examiner's finger did or did not touch the ILP muscle by simulating the performed palpation. Palpatory findings were supplemented by 1-dimensional measurements for determination of topographic relations of the ILP muscle within the infratemporal fossa. For statistical analysis, sensitivity, specificity, and negative and positive predictive values of the palpation technique were calculated. Interexaminer agreement was estimated with the kappa value. RESULTS: In 86 of 106 dissected specimens, a superficial fascicle of the medial pterygoid muscle was found in direct proximity to the ILP muscle. In these cases, a residual distance of 7.8 +/- 3.2 mm remained between the ILP muscle and buccinator fascia indented by the tip of the examiner's finger. In 10 of 20 specimens with an absent superficial fascicle, the finger was able to reach the ILP muscle. CONCLUSION: It is recommended that the ILP muscle palpation technique should no longer be considered as a standard clinical procedure because it is nearly impossible to palpate the ILP muscle anatomically and because the risk of false-positive findings (by palpation of the medial pterygoid muscle) is high. PMID- 10793388 TI - Evaluation of impression accuracy for osseointegrated implant supported superstructures. AB - STATEMENT OF PROBLEM: An often-debated issue still exists concerning implant impression techniques, whether to splint impression copings. Different configurations are available for these copings for a variety of manufacturers' implant systems. PURPOSE: This study evaluated and compared 4 impression techniques in terms of their dimensional accuracy to reproduce implant positions on working casts. MATERIAL AND METHODS: A master model was designed to simulate a clinical situation. Impressions were made using 4 techniques: (1) tapered impression copings not splinted; (2) squared impression copings not splinted; (3) squared impression copings splinted with autopolymerizing acrylic resin; and (4) squared impression copings with a lateral extension on one side not splinted. Reference points machined onto the master model and onto special healing abutments were compared after abutments were transferred to casts using the 4 techniques. Measurements were made using a Reflex microscope, capable of recording in the x-, y-, and z-dimensions. RESULTS: The dimensional accuracy was high and, although statistically significant (P =.022; power > 80%), a maximum distortion difference of only 0.31% was registered. CONCLUSION: The dimensional accuracy of all the techniques was exceptional and the observed differences can be regarded as clinically negligible. PMID- 10793390 TI - In vitro evaluation of the strength of the conical implant-to-abutment joint in two commercially available implant systems. AB - STATEMENT OF PROBLEM: The cone-screw abutment has been shown to diminish micromovement, reducing the burden of component loosening and fracture. However, it is unclear whether the conical taper and joint design influence strength of the interface, with respect to unfavorable bending moments. PURPOSE: This comparative study evaluated the resistance to bending for the ITI Straumann and Astra Tech ST implant systems using an 8- and 11-degree internal cone, respectively. MATERIAL AND METHODS: Assembled units from each system were mounted in a 3-point bending apparatus. High load tests were performed, 4 mm from the joint, and bending moments necessary to induce first point of plastic deformation and ultimate failure were measured. All units were inspected to determine the critical zone of failure. RESULTS: Bending moments necessary to induce first point of plastic deformation were considered well above that expected in clinical function for both systems. However, the critical zones of failure differed in that the solid Astra abutment deformed before the cone joint with its 11-degree taper and smooth transition into the neck of the screw, preventing screw fracture. By contrast, all ITI screws fractured at the head of the screw where it met the base of the 8-degree cone. It is unclear which aspects of the joint design were responsible for the difference observed in mode of failure or if it was a direct result of the experimental design. CONCLUSION: For clinically relevant levels of bending moment, no problems were anticipated with respect to component failure for either system. PMID- 10793389 TI - In vitro comparison of master cast accuracy for single-tooth implant replacement. AB - STATEMENT OF PROBLEM: The inaccuracy in transferring the position of the hexagonal head of a single implant to the working cast can result in a final single tooth crown, which clinically may present occlusal and/or interproximal contacts that are different from those contacts on the master cast obtained by the technician. PURPOSE: This in vitro study evaluated the accuracy of the master casts obtained using square pick-up impression copings for single-tooth replacement. Copings used were (1) copings as sold by the manufacturer, and (2) copings modified by sandblasting and coating with impression adhesive their roughened surfaces before final impression procedures. MATERIAL AND METHODS: A polymeric resin model with a standard single implant was used to simulate a clinical situation. A group of 20 impressions were made using nonmodified impression copings; a second group of 20 impressions were fabricated with modified copings. Master casts fabricated for both groups were analyzed to detect rotational position change of the hexagon on the implant replicas in the master casts in reference to the resin model. RESULTS: The rotational position changes of the hexagon on implant replicas were significantly less variable in the master casts obtained with the modified impression copings than in the master casts achieved with the nonprepared copings. CONCLUSION: Improved precision of the impression was achieved when the adhesive-coated copings were used. PMID- 10793391 TI - Physical properties of a silicone prosthetic elastomer stored in simulated skin secretions. AB - STATEMENT OF PROBLEM: Facial prostheses worn over an extended time are exposed to various environmental factors, including sebaceous oils (sebum) and perspiration. PURPOSE: This study investigated the physical properties of tensile strength and modulus, elongation, tear strength, hardness, weight, and color change, of a silicone facial elastomer after immersion for 6 months in simulated sebum and perspiration at 37 degrees C. MATERIAL AND METHODS: Specimens made of Episil silicone elastomer were immersed in simulated alkaline or acidic perspiration as well as in sebum. Tensile and tear tests were conducted according to ISO specifications no. 37 and 39, respectively, in a Monsanto testing machine. Shore A hardness measurements were run according to ASTM D 2240. Weight changes were followed at 5, 15, 30, and 180 days, and color changes were determined in the CIE LAB system using a tristimulus colorimeter. RESULTS: An improvement of mechanical properties for specimens immersed in acidic perspiration was attributed to facilitation of the propagation of cross-linking reaction during aging of the silicone samples. Some weight increase was observed for the specimens immersed into the aqueous solution, whereas for those immersed in sebum, weight loss was recorded, probably because of extraction of some compounds. In this latter case, the color change was lower than that corresponding to simulated perspiration. CONCLUSION: The silicone specimens aged for a period, which simulates 1.5 years of clinical service, showed minimal changes with respect to the properties studied. PMID- 10793392 TI - Fabrication of a custom silicone tracheostomal prosthesis. AB - Patients with a tracheostomy stoma experience compromised speech due to the associated changes in airflow patterns. Prosthetic obturation of the stoma restores the normal airflow patterns required for proper speech. Standard stent tubes may not adequately obturate the defect and may be uncomfortable to wear. The fabrication of a custom silicone tracheostomal prosthesis, incorporating a speaking valve, provides proper obturation of the defect and improved patient comfort, fit, and function. This article presents the procedures used to make a functional impression of a tracheostomy stoma and to fabricate a custom tracheostomal prosthesis. PMID- 10793393 TI - Creating a vertical stop for interocclusal records. AB - This article describes a method of making an accurate interocclusal record when the most distal tooth is an abutment of a fixed partial denture. The method uses conical stops, prepared in the enamel of the abutment or made of composite or a metal core covered with composite, to maintain the vertical dimension of occlusion and to act as a third point of reference for a stable occlusal relationship when occluding the definitive casts. This article also explains how to use the cones and how to make each variety. PMID- 10793394 TI - Simplified procedure for replacement of retentive clips in a bar-retained implant overdenture. AB - Replacing the retentive elements in a bar-retained overdenture can be a difficult and time-consuming procedure. This article describes an impression procedure that allows the matrices in a bar-retained implant overdenture to be replaced in the laboratory using a heat-cured rebase procedure. Long waxing screws are used, which allows the bar to be removed in the rebase impression and incorporated in a new master cast. After completion of the laboratory phase, the bar is screwed back onto the implant abutments and the denture, with new retentive matrices, is inserted. PMID- 10793395 TI - Procedure for reducing the vertical height of a hollow box interim obturator. PMID- 10793396 TI - Immediate repair for a fractured anterior tooth in a removable prosthesis. PMID- 10793397 TI - Diabetes mellitus in Asia. PMID- 10793398 TI - Obesity is the key determinant of cardiovascular risk factors in the Hong Kong Chinese population: cross-sectional clinic-based study. AB - OBJECTIVES: To examine the interrelationships between obesity and various cardiovascular risk factors, and to investigate the relative importance of insulin and obesity in their associations with various pathophysiologies. DESIGN: Cross-sectional clinic-based study. SETTING: Medical clinics at a university teaching hospital, Shatin, Hong Kong. PARTICIPANTS: A heterogeneous cohort of 767 Hong Kong Chinese subjects with a mean age of 43 (standard deviation, 14) years. MAIN OUTCOME MEASURES: Body mass index, waist circumference, plasma insulin, insulin resistance index, fasting plasma glucose and lipid levels, blood pressure, and 24-hour urinary albumin excretion. RESULTS: Pathophysiological abnormalities and risk factors are frequently clustered to varying degrees. Compared with the control subjects, patients with at least one component of the metabolic syndrome were more obese, hyperinsulinaemic, insulin resistant, hyperglycaemic, hypertensive, dyslipidaemic, and albuminuric (all variables, P<0.001). Increasing degrees of body mass index, waist circumference, plasma insulin level, and insulin resistance index were associated with an increasing number of risk factors after adjusting for age and sex (all variables, P<0.02). Multiple regression analysis showed that obesity, as reflected by either the body mass index or waist circumference, had a closer association than plasma insulin with the fasting plasma glucose concentration, blood pressure, and high-density lipoprotein-cholesterol and triglyceride concentrations. Using 19.0-20.9 kg/m(2) as the reference body mass index interval, the lowest cardiovascular risk was associated with a body mass index of <23.0 kg/m(2). There was an increased risk of 3.1 and 5 times when the body mass index was 23. 0-24.9 kg/m(2) and > or =25 kg/m(2), respectively. CONCLUSIONS: Obesity, hyperinsulinaemia, and insulin resistance are characteristic features of Hong Kong Chinese patients who have various components of the metabolic syndrome. Obesity has a greater effect than plasma insulin on various pathophysiologies. PMID- 10793399 TI - Patterns of referral to the paediatric specialist clinic of a regional hospital: descriptive study. AB - OBJECTIVE: To study the patterns of referral to a paediatric specialist out patient clinic. DESIGN: Descriptive study. SETTING: Regional hospital, Hong Kong. PATIENTS: The 1985 referral letters of patients who were referred to the paediatric specialist clinic during 1998 were studied. MAIN OUTCOME MEASURES: Common referral diagnoses according to sources of referral. RESULTS: The common sources of referral were maternal and child health centres (34.7%), accident and emergency departments (26.9%), the Student Health Service (12.9%), private practitioners (10.5%), and general out-patient clinics (9.8%). The common referral diagnoses from maternal and child health centres (n=689) were growth problems (37. 7%), heart murmurs (16.8%), and neonatal jaundice (10.9%). Asthma or suspected asthma constituted the majority of referrals from accident and emergency departments (227/533; 42.6%). Deviations in growth (41. 4%), problems regarding puberty (13.7%), apparent heart problems (13. 3%), and nocturnal enuresis (11.7%) were the common referral diagnoses from the Student Health Service (n=256). CONCLUSIONS: Studying referral patterns from various sources is helpful in organising medical services and identifying training needs. PMID- 10793400 TI - Failure patterns and factors affecting prognosis of salivary gland carcinoma: retrospective study. AB - OBJECTIVES: To investigate the failure patterns and the prognostic factors following postoperative radiotherapy for salivary gland carcinoma. DESIGN: Retrospective study. SETTING: University teaching hospital, Hong Kong. PATIENTS: Fifty patients who had non-disseminated salivary gland carcinoma and who received primary treatment from 1984 through 1993. MAIN OUTCOME MEASURES: Demographic data, cancer T- and N-stages, histological type, site of origin, completeness of surgery, whether postoperative radiotherapy was given, and the clinical outcome. RESULTS: Two (4%) patients had been treated with radiotherapy alone, six (12%) had undergone radical resection alone, and 42 (84%) had been radically treated by using both modalities. The 5-year overall survival and relapse-free survival rates were 78.4% and 63.1%, respectively. The free from local failure and free from distant metastasis rates at 5 years were 77.2% and 72.8%, respectively. The N-stage was a significant prognostic factor. The site of the primary tumour, T stage, completeness of surgery, and use of postoperative radiotherapy were not significant independent prognosticators; however, among the T-stage tumours, the b-substage carcinomas had significantly fewer local failures (P=0.040) and better survival rates (P=0.038) than the a-substage carcinomas. There were seven (14%) locoregional failures without distant metastasis, seven (14%) cases of distant metastasis without locoregional failures, and four (8%) locoregional failures preceding distant metastasis; isolated regional relapse was rare (1/50; 2%). All regional failures (5/50; 10%) occurred ipsilateral to the primary lesion. There were no deaths due to lymphoepithelioma-like carcinoma or acinic cell carcinoma. CONCLUSIONS: The N-stage is the main prognostic factor of overall survival, relapse- and metastasis-free recovery, and success of treatment for salivary gland carcinoma. Optimal locoregional treatment can help reduce distant metastasis, and the maximal use of postoperative radiotherapy may contribute to improved locoregional control. Elective ipsilateral neck radiotherapy is indicated for lymphoepithelioma-like carcinoma. PMID- 10793401 TI - Autologous bone marrow transplantation for patients with acute myeloid leukaemia: prospective follow-up study. AB - OBJECTIVE: To study the use of autologous bone marrow transplantation to treat acute myeloid leukaemia when complete remission had been achieved and when no human leukocyte antigen matched related donor was available. DESIGN: Prospective follow-up study. SETTING: Government hospital, Hong Kong. PATIENTS: Eight patients (median age, 34 years [range, 16-45 years]) with acute myeloid leukaemia in whom complete remission had been achieved. INTERVENTION: Conditioning regimen of carmustine, amsacrine, etoposide VP-16, cytarabine, and infusion of unpurged marrow. MAIN OUTCOME MEASURES: Median time taken to reach neutrophil and platelet counts of > or =0.5 x 10(9) /L and > or = x 10(9) /L, respectively; mortality and relapse rates; and follow-up regimens used. RESULTS: Engraftment was successfully achieved in all patients and there were no early procedure-related mortalities. The median times required to reach a neutrophil count of > or =0.5 x 10(9) /L and a platelet count of > or =20 x 10(9) /L were 30 days (range, 18-36 days) and 38 days (range, 15-53 days), respectively. The median duration of hospital stay was 37 days (range, 25-43 days). Two patients died of a relapse of leukaemia at 6 and 9 months post-transplantation. Two patients experienced relapses: one at 8 months post-transplantation, for which conventional chemotherapy was restarted, and one at 18 months; treatment with all-trans-retinoic acid and conventional chemotherapy achieved a third complete remission in the latter patient, who had acute promyelocytic leukaemia. Continuous remission has been achieved in four of the eight patients after a median follow-up duration of 26 months (range, 6-43 months). CONCLUSION: Autologous bone marrow transplantation is an acceptable treatment for patients with acute myeloid leukaemia who lack a human leukocyte antigen-matched related donor. PMID- 10793402 TI - The epidemiology of diabetes mellitus in the Asia-Pacific region. AB - The Asia-Pacific region is at the forefront of the current epidemic of diabetes. There are currently more than 30 million people with diabetes in the Western Pacific region alone. The World Health Organization predicts that this number will rise dramatically by the year 2025, by which time India and China may each face the problem of dealing with 50 million affected individuals. The problem in the region results from a combination of large population size with rapidly rising prevalence rates, particularly of type 2 diabetes mellitus. Although much heterogeneity exists, rising prevalence rates are being seen throughout the region and appear to be closely associated with westernisation, urbanisation, and mechanisation. The risk for diabetes appears to result from a combination of genetic predisposition and lifestyle change. The most important lifestyle changes relate to changes in dietary habits and physical activity and diabetes risk, particularly in younger individuals, is associated with the development of obesity and particularly central obesity. In some populations, for example Chinese, the relationship between diabetes and weight gain begins to appear at levels of body weight that would not be conventionally regarded as representing obesity. The increasing trend for type 2 diabetes to develop in young people is of particular concern. In children and adolescents in some parts of the region, type 2 diabetes now outnumbers type 1 diabetes by a ratio of 4:1. In view of the severity of the long-term complications of diabetes, the health consequences of this epidemic will become increasingly devastating and threaten to overwhelm the health care systems in the most vulnerable countries. There is an urgent need for prioritisation of diabetes as a key issue by governments throughout the region. Diabetes prevention programmes can be justified on economic, as well as humanitarian grounds. At the level of primary prevention, such programmes can be linked to other non-communicable disease prevention programmes which also target lifestyle-related issues. PMID- 10793403 TI - Diagnosing diabetes mellitus in the Asian population. AB - Western guidelines have been used to diagnose diabetes mellitus in Asia. The increased availability of data from Asian populations, however, has made it apparent that modifications to western guidelines are needed when they are used in Asia. Both the American Diabetes Association and the World Health Organization have recently modified their diagnostic criteria for diabetes mellitus. The implications of these new criteria in Asia are discussed in this paper. The significance of using fasting plasma glucose measurements and/or oral glucose tolerance tests in the diagnosis of diabetes mellitus is analysed. A simple approach to diagnose diabetes mellitus in the Hong Kong Chinese population is also suggested. PMID- 10793404 TI - Diabetic complications and their implications on health care in Asia. AB - Diabetes mellitus is a growing health problem in the Asia-Pacific region. The acute and chronic complications of diabetes mellitus are major causes of hospital admissions, blindness, renal failure, amputations, stroke, and coronary heart disease in this region. Compared with the general population, the annual per capita health care expenditure is estimated to be four-fold for people with diabetes. Recent prospective studies have provided unequivocal evidence for the crucial role of prolonged hyperglycaemia in the development of chronic diabetic complications. Although the aetiology of hyperglycaemia-induced damage of the kidneys, eyes, nerves, and arteries still remain to be elucidated, observational and interventional studies show that the occurrence and progression of these complications can be prevented by the optimal control of blood glucose, hypertension, and dyslipidaemia. Lifestyle changes such as weight control, increased physical exercise, and smoking cessation are also potentially beneficial in preventing diabetes mellitus and coronary artery disease. Furthermore, the morbidity and mortality caused by diabetes mellitus can be reduced by secondary prevention through regular screening, early detection, and appropriate treatment of chronic complications. Improved diabetes education is needed among health professionals as well as the general and diabetic populations. Government and public health officials should be mindful of the economic impact of this major health problem so that adequate health care resources can be allocated for the primary and secondary prevention of diabetic complications. PMID- 10793405 TI - Genetics of type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a heterogeneous disease that is caused by both genetic and environmental factors. Only a minority of cases of type 2 diabetes are caused by a single-gene defect, such as maturity-onset diabetes of youth (mutated MODY gene), syndrome of insulin resistance (insulin receptor defect), and maternally inherited diabetes and deafness (mitochondrial gene defect). The genetic component of the more common form of type 2 diabetes is probably complex and involves the interactions of multiple genes and environmental factors. The candidate gene approach has identified several genes that regulate insulin signalling and secretion, but their contributions to diabetes are small. Recent genome scan studies have been conducted to identify major susceptibility loci that are linked with type 2 diabetes. This information would provide new insights into the identification of novel genes and pathways that lead to this complex disease. PMID- 10793406 TI - Heterogeneity of diabetes mellitus in the Hong Kong Chinese population. The Chinese University of Hong Kong-Prince of Wales Hospital Diabetes Research and Care Group. AB - Diabetes mellitus is a massive public health problem that has major socio economic implications. There are now local and international data that confirm the high prevalence of diabetes in Chinese populations living in affluent societies such as Hong Kong. The heterogeneity of diabetes mellitus, especially among patients with a young onset of the disease, is being increasingly recognised. Genetic factors appear to be a particularly important factor in these young diabetic patients, who face a long duration of disease. Understanding the pathogenesis and genetic basis of diabetes mellitus and its complications are of fundamental importance to improving the diagnosis and treatment of this heterogeneous disease. Despite its complications, diabetes mellitus is a very treatable and preventable disease. The challenge lies in the effective delivery of quality and affordable care to high-risk individuals. To this end, these concerted efforts among academics, clinicians, scientists, health care professionals, administrators, and policy makers are required to defuse this health care time bomb. PMID- 10793407 TI - Recent advances in the management of depression and psychopharmacology. AB - OBJECTIVE: To review the recent advances in the management of depression and psychopharmacology. DATA SOURCES: Medline and non-Medline literature search. STUDY SELECTION: The following key words were used: depression/therapy, depressive disorders, antidepressant, psychopharmacology, and mental health services. Years of study: 1988 to 1998. DATA EXTRACTION: Original articles, review papers, meta-analyses, and relevant book chapters were reviewed. DATA SYNTHESIS: Recent advances in research on depression have confirmed that it is a common, recurrent, and disabling medical disorder. The latest epidemiological studies from the United States suggest that its lifetime prevalence is more than 17%, while a lower, but still substantial, proportion of Chinese people have the same disorder. The highly recurrent nature and conspicuous morbidity of depression call for the continuation of antidepressant treatment beyond the initial resolution of symptoms. For the first episode of depression, 6 to 9 months of adequate antidepressant treatment is indicated. For patients with recurrent depression, maintenance therapy for several years is needed. The increasingly widespread use of selective serotonin re-uptake inhibitors and other newer antidepressants has increased the direct drug cost, but the total health care expenditure may not be raised and may even be reduced. CONCLUSION: Proper recognition and management of depression at both the clinical and health care policy levels are urgently needed. PMID- 10793408 TI - Using the comprehensive geriatric assessment technique to assess elderly patients. AB - OBJECTIVE: To review the concept, components, and characteristics of the Comprehensive Geriatric Assessment technique. DATA SOURCES: Medline and non Medline literature search. STUDY SELECTION: The following key words were used: Comprehensive Geriatric Assessment; all available years of study were reviewed. DATA EXTRACTION: Studies that assessed the Comprehensive Geriatric Assessment technique's benefits were examined. DATA SYNTHESIS: By using the Comprehensive Geriatric Assessment programme, accurate diagnoses can be made, treatable illness can be screened for, therapeutic plans can be formulated, and the optimal placements of patients can be achieved. Assessment should be performed at each level of geriatric care; various well-validated scales are used to measure the activity of daily living of patients. The Comprehensive Geriatric Assessment programme can improve functional status; reduce the use of medications, nursing homes, and medical services; and reduce mortality rates. Most studies confirm that a successful programme requires careful patient targeting, implementation of the programme by attending physicians, and patient adherence to the recommendations made. CONCLUSION: A well-targeted Comprehensive Geriatric Assessment programme and the control of patients' adherence to recommendations are effective in improving the well-being of elderly patients. PMID- 10793409 TI - Detecting asthma and bronchial hyperresponsiveness in children. AB - The prevalence of asthma in children is increasing worldwide. Although the features of asthma are well documented, defining asthma remains a problem. The clinical definition of asthma does not take into account the concept of airway inflammation. A broader definition that incorporates the inflammatory process, reversibility of airway obstruction, and airway responsiveness needs to be more widely adopted. Bronchial hyperresponsiveness is one of the key features in asthma and it can be documented by using pharmacological or non-pharmacological means. The latter appears to be a more physiological test and more acceptable to children. This article gives an overview of the features of asthma and bronchial hyperresponsiveness and shows how various non-pharmacological bronchial challenge tests can help identify bronchial hyperresponsiveness and thus help diagnose asthma in children. PMID- 10793410 TI - A statement for health care professionals on type 2 diabetes mellitus in Hong Kong. Diabetes division, Hong Kong Society for Endocrinology, Metabolism, and Reproduction. AB - OBJECTIVE: To issue a statement for health care professionals on type 2 diabetes mellitus in Hong Kong. PARTICIPANTS: The Diabetes Division was established under the auspices of the Hong Kong Society for Endocrinology, Metabolism, and Reproduction. The Division consists of medical and paediatric specialists, as well as health educators, including nurses, podiatrists, and dietitians who have a particular interest in diabetes. EVIDENCE: The statement was based on evidence from the available scientific literature on diabetes management from Hong Kong and overseas. CONSENSUS PROCESS: The draft statement was prepared on 16 February 2000 by a working group of diabetologists working in the public and private sectors. It was presented to the Council of the Diabetes Division on 7 March 2000 and approved by the Council of the Hong Kong Society for Endocrinology, Metabolism, and Reproduction on 11 March 2000. CONCLUSIONS: Type 2 diabetes mellitus affects people of all ages and is a massive public health problem. The criteria used to diagnose diabetes mellitus have recently been revised. Physicians require increased vigilance to screen for glucose intolerance in individuals who have risk factors for type 2 diabetes mellitus. There are now recommended cost-effective procedures for the optimal management of type 2 diabetes mellitus, which emphasise regular monitoring, the control of both diabetes and associated risk factors, and self-management. To address this public health problem, concerted efforts by health care professionals and public bodies are needed to increase levels of awareness and improve the standard of care. PMID- 10793411 TI - Use of hyperbaric oxygen therapy in Hong Kong. AB - The Recompression Treatment Centre on Stonecutters Island has been operating in Hong Kong for more than 5 years and has been used to treat a variety of diving related and other conditions by means of hyperbaric oxygen therapy. Up to the end of December 1997, 295 treatment sessions had been conducted for 39 patients. This article reviews the usefulness of and indications for hyperbaric oxygen therapy. PMID- 10793412 TI - Management of carbon monoxide poisoning using oxygen therapy. AB - The management of carbon monoxide poisoning requires an accurate assessment of the extent of blood oxygenation. Measuring the fractional oxyhaemoglobin content by using co-oximetry gives a true picture of the oxygen-carrying capacity of blood in the presence of carboxyhaemoglobin. The use of readings from pulse oximetry or a standard blood gas analyser is insufficient and can be misleading. We report on a case of carbon monoxide poisoning to illustrate this potential pitfall. PMID- 10793413 TI - Polycystic disease: a rare indication for combined liver and kidney transplantation. AB - We report on a 52-year-old woman who presented with polycystic disease. Both of her kidneys had been removed and she had undergone one failed kidney transplantation. She had severe symptoms from the polycystic liver. The diseased liver and kidney were both treated successfully by performing a combined liver and kidney transplantation. PMID- 10793414 TI - Use of oral valaciclovir in a 12-year-old boy with herpes simplex encephalitis. AB - We report on a 12-year-old boy with herpes simplex encephalitis, in whom a severe localised skin reaction developed following the infusion of intravenous acyclovir. Oral valaciclovir was given as continuation therapy to complete the 3 week course of antiviral treatment and resulted in complete recovery without side effects. This report illustrates the advantage of using the polymerase chain reaction to diagnose herpes simplex encephalitis and the potential use of newer antiviral agents, such as valaciclovir, as continuation therapy in the management of the infection. The higher oral bioavailability of newer antiviral agents allows part of the extended treatment period of patients with herpes simplex encephalitis to be carried out as an ambulatory oral regimen. PMID- 10793415 TI - [Current concept in pathogenesis of migraine and its treatments]. PMID- 10793416 TI - [Deep brain stimulation therapy: control of human brain function by chronically implanted electrodes]. AB - During the last decade, it has become increasingly clear that deep brain stimulation(DBS) is useful for controlling various neurological disorders, including intractable pain and involuntary movements. The development of an implantable and programmable pulse-generator has greatly facilitated the clinical application of DBS therapy. The history and clinical data of DBS therapy currently available from the literature, together with our own cases treated by DBS, are reviewed. DBS therapy has many advantages over conventional neuroablative therapies, including reversibility of the procedure, controllability and relative selectivity of the effects, and possible plastic effects on neural function. Further evolution of the stimulation system, such as through integration with neuromonitoring devices, would open up a new field of restorative neurology and neurosurgery. PMID- 10793417 TI - [Evaluation of cerebral metabolism by multi-voxel proton magnetic resonance spectroscopy imaging in chronic unilateral internal carotid artery occlusion]. AB - Proton magnetic resonance spectroscopy(1H-MRS) has less been used to analyze cerebral metabolism in ischemic lesions compared to single photon emission computed tomography or positron emission computed tomography. Recent advances in magnetic resonance imaging apparatus and the related software have made possible obtaining multi-voxel 1H-MRS in a single study. We examined multi-voxel 1H-MRS in patients with unilateral internal carotid artery(ICA) occlusion to study the relationship between cerebral metabolism and cerebral blood flow. Fifteen patients(male 11; female 4, 47-76; average 67.1 year-old) with chronic unilateral ICA occlusion and without any marked infarction were studied. 1H-MRS was obtained using a 1.5 T Siemens Magnetom Vision scanner. Multi-voxel spectra were recorded using a SE-2 D-CSI sequence(TR/TE = 1500/135 ms). The volume of interest was 90 x 90 x 20 mm3, placed axially above the lateral ventricle. The single voxel size was 10 x 10 x 20 mm3. N-acetyl aspartate/creatine ratios(NAA/Cr) were calculated on each voxel and were averaged in view of the cortex and the white matter. The regional cerebral blood flow(CBF) was measured by Xenon-CT method. Eight patients were also examined by acetazolamide challenge to evaluate the cerebrovascular reserve capacity. NAA/Cr ratios in normal subjects were 1.905 +/- 0.090(mean +/- standard deviation) in the cortex and 2.183 +/- 0.258 in the white matter in 40's(n = 6), 2.046 +/- 0.166 in the cortex and 2.039 +/- 0.288 in the white matter in 60's(n = 5). The study revealed 7 patients with normal NAA/Cr ratio and CBF, 5 with reduced NAA/Cr ratio and normal CBF, and 3 with reduced NAA/Cr ratio and CBF in the affected cortex. A low correlation coefficient of 0.46 was noted between NAA/Cr ratio and the cerebrovascular reserve capacity calculated by acetazolamide challenge in the affected cortex. In the range of less than +10%(lower limit) in percentile change of regional CBF after acetazolamide injection, NAA/Cr ratio was distributed between 1.600 and 2.044, which were normal or slightly under the lower limit(mean-2 x standard deviation). Multi voxel 1H-MRS is useful for the evaluation of cerebral metabolism, because it enables to quantify different chemicals in many fields at one time and to compare its distribution with regional CBF. In patients with unilateral ICA occlusion, NAA/Cr ratio of the affected cortex varies depending on the collateral circulation and the contralateral ICA lesions. The Extracranial-Intracranial Bypass should be considered if the case with unilateral ICA occlusion reveals reduced CBF and normal or slightly decreased NAA/Cr ratio in the affected cortex. PMID- 10793418 TI - [Persistent primitive trigeminal artery presenting with Weber's syndrome: report of a case with three-dimensional CT angiographic evaluations]. AB - We report a rare case of persistent primitive trigeminal artery(PPTA) presenting with brain stem infarction known as Weber's syndrome, and document its unique findings of three-dimensional CT angiography(3 D-CTA). A 69-year-old woman was admitted to our hospital because of gait disturbance and blepharoptosis on the right eye. Neurological examination on admission revealed the right oculomotor nerve palsy, left hemiparesis and dysarthria, all of which indicated the signs and symptoms of Weber's syndrome. Initial CT scan revealed no abnormality, but a subsequent 3 D-CTA demonstrated the PPTA originating from the right internal carotid artery penetrate into the clivus directly to the distal basilar artery, on top of which a small saccular aneurysm was incidentally visualized. Right internal carotid angiograms showed the PPTA run between the cavernous segment of the internal carotid artery and the distal portion of the basilar artery with the filling of both the posterior cerebral and superior cerebellar arteries. However, the proximal portion of the basilar artery was visualized through the right vertebral artery and there was no blood flow to its distal portion. Evidence of infarction was finally confirmed at the right midbrain and thalamus by the MRI performed 5 days after the onset. With a conservative treatment including physical therapy, the patient recovered well from the deficits and could walk by herself with a cane. With regard to the pathogenesis of vertebrobasilar insufficiency in a patient with PPTA, it is generally considered that microembolus from an atherosclerotic carotid artery may be its cause because of the presence of direct communication between the anterior and posterior circulations. In the present case, however, this mechanism may not be applied since there was no evidence of atherosclerotic plaque or stenotic lesions on the carotid arteries. Alternatively, an embolic occlusion may have occurred in the paramedian branches of the posterior cerebral artery since a dilated PPTA itself, which resembled fusiform-aneurysm in appearance, may become the origin of microembolus. PMID- 10793419 TI - [A case of hematoma in cisterna magna after mild head injury]. AB - An 18-year-old man suffered severe neck pain, signs of meningeal irritation, and dysesthesia of his left breast and left arm after mild head injury incurred in a traffic accident. On the next day, magnetic resonance imaging(MRI) revealed a hematoma in the cisterna magna, and it extended into the posterior spinal subdural space. Eight days after admission, MRI demonstrated the hematoma on the right posterior aspect of the spinal cord at the level of craniovertebral junction. These MRI findings suggested a ruptured bridging dorsal vein from the spinal cord to the occipital or marginal sinus. The usefulness of MRI for diagnosis of localized hematoma around the craniovertebral junction and the mechanism of this hemorrhage are discussed. PMID- 10793420 TI - [A case of glioblastoma multiforme which indicated the early stage on brain MRI]. AB - A 57-year-old male was urgently carried to our hospital because of sudden loss of consciousness, lasting about 10 minutes. He had resumed consciousness before he arrived at our hospital. Neurologically, he had mild muscle weakness of the right arm. Deep tendon reflexes in the right upper extremity were reduced. In high level functions, speech disturbance, dysgraphia (disturbed ability to write Hiragana), and constructive apraxia were noted. A brain MRI upon admission showed a poorly demarcated, high signal intensity area in the cortical and subcortical layers of the left temporal and parietal lobes. This was visible on T 2 weighted images(T 2 WI), although no abnormalities were visible on T 1 weighted images(T 1 WI). No contrast enhancement was effected by Gd-DTPA. The patient was therefore suspected of having a tumor or degenerative disease and was monitored closely. About 4 months later after onset, his symptoms became aggravated, and brain MRI disclosed a marked low signal intensity area on T 1 WI and a heterogeneous high signal intensity area on T 2 WI. The abnormal signal intensity area was surrounded by extensive edema and mass effect. Ring-shaped, irregular, contrast enhanced areas were also visible. Cerebral angiography revealed a poorly demarcated tumor stain in the area supplied by the middle cerebral artery. The tumor was removed surgically and was histopathologically rated as glioblastoma multiforme(GBM). Because this case represents a valuable example of early stage of GBM, it will be discussed in this paper, along with differential diagnoses. PMID- 10793421 TI - [A case of alcoholic multiple nervous system degeneration]. AB - Chronic ethyl alcohol abuse is associated with different types of neurological involvement. We report a 51-year-old woman with alcoholic encephalopathy, neuropathy and autonomic dysfunction. After the alcohol abuse of about thirty years, gait disturbance, dysphagia and dysarthria progressively worsened. We thought that the disease was caused by poor nutrition due to chronic alcohol abuse and vitamin B1, B12 deficiency. Her neurological symptoms and signs improved after discontinuation of alcohol and nutritional treatment. PMID- 10793422 TI - [A case of asymptomatic meningioma at left frontal convexity: evaluation using of WAIS-R at preoperation and postoperation]. AB - The authors reported a case of 39-year-old man with incidentally found meningioma whose postoperative WAIS-R scores were improved significantly. He got a minor head injury run in a car collision and showed no neurological findings on admission. Computed tomography revealed no traumatic abnormalities but left frontal convexity meningioma. Total removal of the tumor was performed 3 weeks after the admission. Preoperative verbal IQ, performance IQ, lower full IQ were 105, 95, 100 respectively, and improved to 113, 112, 114 postoperatively. An asymptomatic patient with incidentally found meningioma could have higher brain function disturbances, which could be improved after its resection, therefore we should operate on such a patient more positively. PMID- 10793423 TI - [Intracranial vertebral artery stenosis successfully revascularised by cerebral angioplasty and stenting in chronic stage: a case report of prompt improvement of the neurological impairment]. AB - A 72-year-old man with a history of hypertension had a left cerebellar infarction and followed by a right cerebellar infarction within about one and a half months after the initial stroke. Brain magnetic resonance images(MRI) showed infarctions in both middle cerebellar peduncles and in the mid-portion of lower pons. Right veretebral artery(VA) terminated in posterior inferior cerebellar artery(PICA). Left intracranial VA has a high-grade eccentric atherosclerotic stenosis(91%) proximal to the left PICA. No collateral circulation was developed from bilateral carotid arteries. Three months after the final ischemic episode, the patient had remained bed ridden and needed a whole assistance for regular daily life because of severe ataxia of four limbs and truncs and of left hemiparesis. The patient and his family gave us informed written consent, then cerebral angioplasty and stenting(CAS) was performed for the left VA stenosis, which was sufficiently dilated. Iodine-123 iodoamphetamine(123I-IMP) single photo emission computed tomography (SPECT) showed hypoperfusion in both cerebellar hemispheres before CAS. Post CAS 123I-IMP SPECT scans demonstrated improvement of the hypoperfusion in the left cerebellar hemisphere. Ataxia of four limbs, left hemiparesis and his will for physical therapy improved in a short period after the treatment. Ten months later, the left VA had a mild stenosis and patient presented mild truncal ataxia and needed less assistance for regular daily life. The present case indicated that improvement of neurological impairment was expected by the endovascular revascularization even in a chronic stage. PMID- 10793424 TI - [Spinal posterior roots enhancement on MRI in a case of Guillain-Barre syndrome]. PMID- 10793425 TI - [Colloid cyst diagnosed effectively by MRI FLAIR]. PMID- 10793426 TI - [A 60-year-old man with numbness and adynamia of the right lower extremity, and finally diagnosed by craniotomic biopsy]. PMID- 10793427 TI - Should lead explantation be the practice standard when a lead needs to be replaced? PMID- 10793428 TI - Long-term outcome of patients with multiple (> or = 3) noninfected transvenous leads: a clinical and echocardiographic study. AB - To prospectively assess the incidence and clinical significance of thromboembolic complications in patients with multiple (> or = 3) noninfected transvenous leads; 48 consecutive patients were evaluated. Half of the patients had two ventricular leads and one atrial lead, 15 patients had two atrial leads and one ventricular lead, while 9 patients had two ventricular and two atrial leads. No additional care was provided except for aspirin (80 mg bid) and annually performed echo Doppler studies. Clinical follow-up included signs and symptoms of subclavian and/or axillary vein thrombosis, the presence of right congestive heart failure, the number of hospital admissions, and death. Echo-Doppler studies assessed the presence of an enlarged right atrium or ventricle, right atrial or ventricular spontaneous contrast, and the presence of tricuspid regurgitation. During a total follow-up of 7.4 +/- 2.2 years there were no differences in the incidence of clinical variables as compared to age-matched controls with DDD pacemakers. The most common complication was transient venous thrombosis (mostly presenting as venous prominence 1-2 weeks after implantation), which was seen in 17% of the study group versus 15% in controls (NS). Cumulative mortality was not different in both groups (13% in the study group vs 15% in controls). No differences were present with respect to hospital admissions (1.1 +/- 0.27/year in the study group vs 1.2 +/- 0.30/year in the controls). In patients with multiple ventricular leads, tricuspid regurgitation on echo-Doppler studies was more frequent (24%) as compared to controls (4%); however, clinical signs of right heart failure were equally distributed. Thus, patients with multiple (> or = 3) noninfected leads have no clinical adverse outcome during long-term follow-up. PMID- 10793429 TI - Efficacy and safety of bipolar sensing with high atrial sensitivity in dual chamber pacemakers. AB - In dual chamber pacemakers, atrial sensing performance is decisive for maintenance of AV synchrony. Particularly, the efficacy of mode switching algorithms during intermittent atrial tachyarrhythmias depends on the sensitive detection of low potential amplitudes. Therefore, a high atrial sensitivity of 0.18 mV, commonly used in single lead VDD pacemakers, was investigated for its efficacy and safety in DDD pacing. Thirty patients received dual chamber pacemakers and bipolar atrial screw-in leads for sinus node syndrome or AV block; 15 patients suffered from intermittent atrial fibrillation. Pace makers were programmed to an atrial sensitivity of 0.18 mV. Two weeks, 3, 9, and 15 months after implantation, P wave sensing threshold and T wave oversensing thresholds for the native and paced T wave were determined. The myopotential oversensing thresholds were evaluated by isometric contraction of the pectoral muscles. Automatic mode switch to DDIR pacing was activated when the mean atrial rate exceeded 180 beats/min. The patients were followed by 24-hour Holter monitoring. Two weeks after implantation, mean atrial sensing threshold was 1.81 +/- 0.85 mV (range 0.25-2.8 mV) without significant differences during further follow-up. Native T wave sensing threshold was < 0.18 mV in all patients. In 13% of patients, paced T waves were perceived in the atrial channel at the highest sensitivity. This T wave sensing could easily be avoided by programming a postventricular atrial refractory period exceeding 300 ms. Myopotential oversensing could not be provoked and Holter records showed no signs of sensing dysfunction. During a 15-month follow-up, 1,191 mode switch events were counted by autodiagnostic pacemaker function. Forty-two of these events occurred during Holter monitoring. Unjustified mode switch was not observed. In DDD pacemakers, bipolar atrial sensing with a very high sensitivity is efficient and safe. Using these sensitivity settings, activation of the mode switch algorithm almost completely avoids fast transmission of atrial rate to the ventricle during atrial fibrillation. PMID- 10793430 TI - P wave and far-field R wave detection in pacemaker patient atrial electrograms. AB - This study was undertaken to develop and test a morphology-based adaptive algorithm for real-time detection of P waves and far-field R waves (FFRWs) in pacemaker patient atrial electrograms. Cardiac event discrimination in right atrial electrograms has been a problem resulting in improper atrial sensing in implantable devices; potentially requiring clinical evaluation and device reprogramming. A morphology-based adaptive algorithm was first evaluated with electrograms recorded from 25 dual chamber pacemaker implant patients. A digital signal processing (DSP) system was designed to implement the algorithm and test real-time detection. In the second phase, the DSP implementation was evaluated in 13 patients. Atrial and ventricular electrograms were processed in real-time following algorithm training performed in the first few seconds for each patient. Electrograms were later manually annotated for comparative analysis. The sensitivity for FFRW detection in the atrial electrogram during off-line analysis was 92.5% (+/- 10.9) and the positive predictive value was 99.1% (+/- 1.8). Real time P wave detection using a DSP system had a sensitivity of 98.9% (+/- 1.3) and a positive predictivity of 97.3% (+/- 3.5). FFRW detection had a sensitivity of 91.0% (+/- 12.4) and a positive predictivity of 97.1% (+/- 4.2) in atrial electrograms. DSP algorithm tested can accurately detect both P waves and FFRWs in right atrium real-time. Advanced signal processing techniques can be applied to arrhythmia detection and may eventually improve detection, reduce clinician interventions, and improve unipolar and bipolar lead sensing. PMID- 10793431 TI - Safety and tolerability of an aggressive tilt table test protocol in the evaluation of patients with suspected neurocardiogenic syncope. AB - Safety and tolerability of a one-step tilt table test with high dose (5 micrograms/min) isoproterenol (ISO) without intermediate stages were evaluated in a symptomatic population of 300 patients referred for clinical syncope, near syncope, or dizziness. ISO has been used as a provocative test but remains controversial. A population of 118 male and 182 female patients with a mean age of 45 (range 5-90) years underwent 300 tests. Heart rate and blood pressure were monitored continuously. A positive test was one in which clinical symptoms were reproduced or hemodynamic criteria met. Patients were initially supine for 5 minutes followed by head upright tilt (HUT) to an angle of 80 degrees for 10 minutes. Negative tests were repeated with an infusion of ISO at a rate of 5 micrograms/min. HUT was positive in 133 (44.3%) of 300 tests. With a 10-minute HUT alone, only 17 (5.7%) of 300 of tests were positive. Of the initial negative tests, 273 of 283 were tested with ISO. With ISO, 116 (42.5%) of 273 were positive. ISO in high dose (5 micrograms/min) was used in 264 of 273 patients, while low dose (1.0-2.5 micrograms/min) was used in 9 of 273 under special circumstances. High dose ISO was tolerated in 164 (62.1%) of 264 patients, reduced in 87 (33%) of 264, and discontinued in 11 (4.2%) of 264. Reasons for reduction included tachycardia (40 patients), nausea (31 patients), chest pain (2 patients), arrhythmia (5 patients), or other (9 patients). Adverse effects resolved within 1 minute of dose reduction. This one-step high dose ISO protocol reproduced neurocardiogenic syncope in symptomatic patients who tested negative without ISO and was safe, tolerated, and expeditious. PMID- 10793432 TI - Permanent transfemoral pacemaker implantation is the technique of choice for patients in whom the superior vena cava is inaccessible. AB - We describe transfemoral pacemaker implantation in three patients in whom pacing via the superior vena cava was not possible or suboptimal. The first was an 88 year-old man with superior vena cava obstruction presenting with fractured epicardial pacing leads. Recent pneumonia increased the risks of a general anesthetic. The second patient was a 57-year-old man who was intolerant of a pectorally sited pacemaker because of the thinness of his anterior chest wall. The third patient was a 69-year-old woman who presented with an infected eroding pectorally sited pacemaker. Scarring secondary to a previous pacemaker infection rendered the contralateral pectoral site inaccessible. Since the subclavian route was inaccessible (case 1) or suboptimal (case 2 and 3), we implanted transvenous pacemakers via the femoral route, which was safe, and effective, during a 6-month follow-up period. PMID- 10793433 TI - The effect of psychological intervention on patients' long-term adjustment to the ICD: a prospective study. AB - This study prospectively examined the role of cognitive behavioral therapy (CBT) in (1) alleviating psychological and somatic distress, and (2) lowering arrhythmic events requiring shocks. Forty-nine of 61 consecutive patients were randomized into therapy (CBT, n = 25) or no therapy (NT, n = 24) and completed a battery of self-report questionnaires at baseline and at 9-month follow-up. CBT was administered at preimplant, predischarge, and at seven routine follow-up visits. Patients were 65 +/- 10 years old, 65% were men, and 92% Caucasian. Eighteen (72%) CBT patients and 18 (75%) NT patients were retained at follow-up. Compared to CBT patients, NT patients reported higher levels of depression (P = 0.046), more anxiety (P = 0.013), more psychological distress (P = 0.015), poorer overall adjustment (P = 0.009), and poorer sexual functioning (P = 0.003). Mean number of shocks did not differ between the CBT and NT groups (2.85 vs 2.30, respectively); however, more patients in the CBT group (61%) than the NT group (33%) received shocks (P = 0.070). At follow-up, a subgroup analysis revealed that the significant differences observed between the CBT and NT groups were attributable to the patients who received shocks in both groups. In conclusion, CBT was associated with decreased depression, decreased anxiety, and increased adjustment for ICD recipients, particularly among those patients receiving shocks. CBT can be administered effectively at routine follow-up visits or transtelephonically with little added inconvenience to the ICD recipient. PMID- 10793434 TI - Chemoreflexsensitivity in patients with survived sudden cardiac arrest and prior myocardial infarction. AB - For evaluation of patients with an increased risk of sudden cardiac death, the analyses of ventricular late potentials, heart rate variability, and baroreflexsensitivity are helpful. But so far, the prediction of a malignant arrhythmic event is not possible with sufficient accuracy. For a better risk stratification other methods are necessary. In this study the importance of the ChRS for the identification of patients at risk for ventricular tachyarrhythmic events should be investigated. Of 41 patients included in the study, 26 were survivors of sudden cardiac arrest. Fifteen patients were not resuscitated, of whom 6 patients had documented monomorphic ventricular tachycardia and 9 had no ventricular tachyarrhythmias in their prior history. All patients had a history of an old myocardial infarction (> 1 year ago). For determination of the ChRS the ratio between the difference of the RR intervals in the ECG and the venous pO2 before and after a 5-minute oxygen inhalation via a nose mask was measured (ms/mmHg). The 26 patients with survived sudden cardiac death showed a significantly decreased ChRS compared to those patients without a tachyarrhythmic event (1.74 +/- 1.02 vs 6.97 +/- 7.14 ms/mmHg, P < 0.0001). The sensitivity concerning a survived sudden cardiac death amounted to 88% for a ChRS below 3.0 ms/mmHg. During a 12-month follow-up period, the ChRS was significantly different between patients with and without an arrhythmic event (1.64 +/- 1.06 vs 4.82 +/- 5.83 ms/mmHg, P < 0.01). As a further method for evaluation of patients with increased risk of sudden cardiac death after myocardial infarction the analysis of ChRS seems to be suitable and predicts arrhythmias possibly more sensitive than other tests of neurovegetative imbalance. The predictive importance has to be examined by prospective investigations in larger patient populations. PMID- 10793435 TI - 24-hour heart rate variability in patients with vasovagal syncope. AB - Since alterations in the autonomic nervous system are thought to play a major role in the pathogenesis of vasovagal syncope, we characterized the chronic autonomic profile of 44 patients with syncope and 20 healthy subjects by means of heart rate variability using 24-hour Holter recordings (time- and frequency domain indexes), and evaluated whether the different types of responses to tilting (vasodepressive versus cardioinhibitory) could be associated with different cardiac autonomic patterns. Twenty-three patients exhibited a positive response to tilting, which was vasodepressive in 11 patients and cardioinhibitory in 12 patients. All vasodepressive patients had a standard deviation of the averages of NN (SDANN) intervals in all 5-minute segments lower than 100 ms. Patients with vasodepressive syncope also had significantly lower values of RMSSD (the 24-hour square root of the mean of the sum of the squares of differences between adjacent normal RR intervals) than those with cardioinhibitory response, and lacked the day-night rhythm of the low frequency/high frequency ratio. However, only SDANN values correctly identified patients with vasodepressive response to tilting. We conclude that (1) the population of patients with vasovagal syncope is heterogeneous, (2) patients with vasodepressive syncope have a peculiar chronic autonomic profile as assessed by 24-hour heart rate variability analysis, and (3) the evaluation of the autonomic profile in 24-hour Holter recordings could be of value in the diagnosis of patients with syncope. PMID- 10793436 TI - Atrial fibrillation induction and determination of atrial vulnerable period using very low energy synchronized biatrial shock in normal subjects and in patients with atrial fibrillation. AB - The atrial vulnerable periods (AVP) for shock induction of atrial fibrillation (AF) in humans have not been clearly defined. Furthermore, the safety and efficacy of using low energy biatrial shock delivered transvenously for AF induction are unknown. We tested the safety and efficacy of using very low energy biatrial shocks, delivered between the right atrium and the coronary sinus for AF induction and used this technique to characterize the AVP in nine controls and nine patients with AF. Thirty-volt and 60-V 3/3-ms biphasic shocks were delivered, starting from 50 ms before the atrial effective refractory period with 20-ms increments until the end of the QRS interval to determine the AVP front, AVP end, and the AVP duration. Successful AF induction could be achieved in eight (89%) of the nine controls and in nine (100%) of the nine patients with AF without any complication. In patients with AF, the AVP front started significantly earlier within the QRS complex, and the AVP duration and the AVP duration/QRS percent ratios were also significantly greater as compared to controls. Furthermore, a higher induction shock energy in patients with AF was associated with an increase in AF inducibility and significantly increased the AVP duration and AVP duration/QRS percent ratio as compared to the controls. This study demonstrated the safe and efficacy of delivering a very low energy biatrial shock during the AVP within the R wave for AF induction. The characteristics of AVP in patients with AF were significantly different from normal subjects. PMID- 10793437 TI - Age related changes in dual AV nodal physiology. AB - Dual atrioventricular nodal (DAVN) physiology has been reported in up to 63% of pediatric patients with anatomically normal hearts, yet atrioventricular nodal reentrant tachycardia (AVNRT) accounts for only 13%-16% of supraventicular tachycardia (SVT) in childhood. The incidence of AVNRT increases with age and becomes the most common form of SVT by adolescence. We investigated the age related electrophysiological responses to programmed atrial and ventricular stimulation in 14 pediatric patients who underwent intracardiac electrophysiological study prior to radiofrequency catheter ablation for AVNRT and who exhibited DAVN physiology. Single atrial and ventricular extrastimuli were placed following drive trains with cycle lengths of 400-700 ms and 350-500 ms, respectively. Six children (mean age 8.2 years, range 5.2-11.5 years) were compared to eight adolescents (mean age 16.6 years, range 13.3-20.7 years). Adolescents were found to have a significantly longer fast pathway effective refractory period (ERP) (median 375 vs 270 ms, P = 0.03), slow pathway ERP (median 270 vs 218 ms, P = 0.04), atrio-Hisian (AH) during AVNRT (median 300 vs 225 ms, P = 0.007), and AVNRT cycle length (median 350 vs 290 ms, P = 0.03). There was a strong trend for the AH measured at the fast pathway ERP to be longer in adolescents than in children (median 258 vs 198 ms, P = 0.055). The AH at the fast pathway ERP was more strongly correlated with baseline cycle length than with age (r = 0.7, P = 0.01 vs r = 0.5, P = 0.7). There was no significant difference in the retrograde VA conduction between adolescents and children. These results demonstrate an age related difference in AV nodal response to programmed atrial stimuli in pediatric patients with DAVN physiology and AVNRT. These differences are consistent with mechanisms that may explain the increased incidence of AVNRT in adolescents compared to children. PMID- 10793438 TI - Identification of the slow conduction zone in idiopathic left ventricular tachycardia. AB - The mechanism of verapamil sensitive idiopathic left ventricular tachycardia (ILVT) is considered to be reentry. However, the nature of the reentry circuit, including the location of the slow conduction zone, is unclear. We sought the local electrical activity that would reflect slow conduction by precise mapping around the tachycardia exit (TE) in nine patients with ILVT (mean age, 28 +/- 10 years) undergoing radiofrequency catheter ablation (RFCA). The TE was defined as the earliest discrete spiky potential (SP) recorded during the tachycardia, or as a complete configuration-matched pacemap 12-lead electrocardiogram (ECG). In all patients, the TE was located at the mid or inferior distal portion of the septum. The SP at the TE preceded the surface QRS by 20 +/- 9 ms. The pacemap score at the TE was 11.4 +/- 0.6 points. In three patients, fractionated potentials (FP) were recorded during the tachycardia. The onset of the FP preceded the surface QRS by 47 +/- 8 ms and was earlier than the SP at the TE (P < 0.01). The sites where an FP was detectable were restricted to a small area, and were at a distance of 14 +/- 4 mm from the TE. The direction of the FP site from the TE was more basal in two patients and inferior in one. Pacemap ECGs at the sites with an FP showed poor matching (9 +/- 1 points), presumably because of predominant capture of the local ventricular muscle rather than an electrically isolated reentry circuit. Successful RFCA was achieved at the site of the FP in all three patients in which one was recorded, and at the TE in the other six patients. The FP, which has been shown to reflect the slow conduction of the ventricular tachycardia circuit in structural heart disease, was also detected in ILVT in the present study, and it is likely to reflect electrical excitation of the distal rim of the slow conduction zone. PMID- 10793439 TI - Serial lead impedance measurements confirm fixation of helical screw electrodes during pacemaker implantation. AB - The purpose of this study was to determine whether serial measurements of helical screw pacemaker lead impedance could reliably confirm electrode fixation in the right atrium and right ventricle. Fixation is generally assessed fluoroscopically, which can be misleading because the myocardium is radiolucent. Alternatively, because the electrical conductivity of blood is greater than that of myocardium, serial measurements of the lead impedance might be expected to show an impedance increase with appropriate fixation of the pacemaker electrode when the electrode becomes embedded in myocardial tissue. Impedance measurements were made during the placement of 23 atrial and 28 ventricular active fixation electrodes in 31 consecutive patients. Impedance measurements were recorded in unipolar and bipolar electrode configurations with the electrode free floating in the chamber, unfixed (with exposed screws) but touching the endocardial surface, and after fixation. No significant impedance differences were found between free floating and unfixed electrode positions. With fixation, the lead impedance increased significantly in the ventricle (P = 0.0001, unipolar and bipolar) and the atrium (P = 0.0069 unipolar and 0.0052 bipolar). Typical increases, reflected by median values, were 197 ohms unipolar and 203 ohms bipolar in the ventricle and 47 ohms unipolar and 53 ohms bipolar in the atrium for electrodes with permanently exposed or retractable screw designs. Comparing serial measurements of lead impedance before and after electrode fixation is a valid electrical method of confirming appropriate fixation of helical screw electrodes. PMID- 10793440 TI - Tilt training: a treatment for malignant and recurrent neurocardiogenic syncope. AB - The treatment of neurocardiogenic syncope is insufficient in many cases. We hypothesized that the repeated exposure of the cardiovascular system to orthostatic stress could have a therapeutic effect on the regulation of cardiovascular reflex mechanisms. We have started a program of tilt training for heavily symptomatic patients. After hospital admission, patients were tilted daily (60-degree inclination), until syncope, or until a maximum of 45-90 minutes. The patients were instructed to continue a program of daily tilt training at home: two 30-minute sessions of upright standing against a vertical wall. No medication was prescribed. A total of 260 tilt table sessions were performed in 42 patients. The first tilt test was positive after 21 +/- 13 minutes. The syncope was cardioinhibitory in 14 cases, vasodepressor in 19, mixed in 9. At the time of hospital discharge, 41 patients could support 45 minutes of head-up tilting. After a mean follow-up time of 15.1 (SD 7.8) months, 36 patients remained completely free of syncope. Syncope still occurred in one patient and presyncope in four patients. One patient died from an extensive myocardial infarction. The abnormal autonomic reflex activity of neurocardiogenic syncope can be remedied by a program of continued tilt training without the administration of drugs. This new treatment has proven to be effective for the vasodepressor and the cardioinhibitory type of syncope. PMID- 10793441 TI - Successful results of a bipolar active fixation lead for atrial application: an interim analysis. AB - Adequate atrial lead performance consists of stable sensing and pacing properties. To evaluate whether the CPI 4269 bipolar lead, covered with mannitol (Sweet Tip), in the atrial position encounters these properties, we performed a prospective study of this lead. After complete dissolution of the mannitol helix, mapping of the atrium to obtain the highest electrogram and lowest threshold was followed by screw-in into the endocardium. Intraoperative measurements were performed and long-term follow-up was scheduled every 6 to 12 months to measure threshold and perform an intracardial electrogram. Between February 1993 and December 1996, a total number of 73 leads in the atrial position in a consecutive series of patients was implanted. Implantation was performed in 28 patients receiving an AAIR and 45 patients a DDDR pacemaker. Reason for pacemaker implantation was a third-degree AV block in 37% of patients, type II second degree AV block in 25%, sick sinus syndrome in 35%, and drug refractory paroxysmal atrial fibrillation following His-bundle ablation in 3%. The intraoperative bipolar atrial electrogram had a mean voltage of 4.25 +/- 2.1 mV. The acute atrial bipolar threshold was 0.63 +/- 0.43 V, and current was 1.35 +/- 0.81 mA at a 1.0-ms pulse duration. The mean acute resistance of the lead was 572 +/- 86 Ohm. After a mean follow-up of 18.3 months, the bipolar intracardial electrogram was 3.37 +/- 2.00 mV, the mean atrial threshold measured at the last outpatient clinic visit was 0.99 +/- 0.74 V and the mean impedance was 640 +/- 127 Ohm. A sensing problem due to traction of the atrial lead occurred in only one patient. Acute and late dislodgement did not occur. The CPI 4269 (Sweet Tip) lead is manufactured with a dissolvable capsule covering the helix tip electrode, permitting a safe passage through the venous system. This interim analysis shows that this lead in the atrial position has favorable acute and chronic results. PMID- 10793442 TI - Effect of rates of perfusion on dominant frequency and defibrillation energy in isolated fibrillating hearts. AB - This study assessed the influence of rates of reperfusion on excitability of the myocardium using dominant frequency (DF) (in Hz) of VF and the relationship of DF to the minimum defibrillation energy (MDE) (in J). Our hypothesis was that increasing flow during reperfusion increases DF that raises MDE. Initially, six Langendorff perfused swine hearts were serially fibrillated and perfusion arrested for 4 minutes followed by reperfusion and defibrillation to establish reproducibility of the model. The epicardial ECG was analyzed for DF. In subsequent studies (n = 8), no flow VF was followed by 1-minute reperfusion at normal flow or 10% flow (low flow) and shocked with increasing energy via epicardial pads until defibrillation. The DF at onset of no flow VF was 9.5 +/- 1.4 and decreased to 3.6 +/- 1.4 after 4 minutes. Reperfusion at normal flow increased the DF of VF compared to low flow after 1 minute (10.8 +/- 1.1 vs 4.5 +/- 1.1 Hz, P = 0.0002) and was associated with increased defibrillation energy requirements (13.5 +/- 5.0 vs 7.3 +/- 6.2 J, P = 0.047). In summary, defibrillation energy requirements are lower when myocardial excitability is reduced during low flow reperfusion. PMID- 10793443 TI - Influence of acute exposure to high altitude and hypoxemia on ventricular stimulation thresholds in pacemaker patients. AB - Cardiac stimulation threshold of implanted pacemakers may be influenced by a variety of endogenous and exogenous factors. High altitude provokes hypoxemia, which may change stimulation thresholds, besides causing other important physiological changes. The aim of our study was to investigate the influence of high altitude on ventricular stimulation thresholds in pacemaker patients. Thirteen patients (10 men; aged 65.5 +/- 4.8 years) with implanted single chamber pacemakers (nine with Pacesetter Regency SR+ with the Autocapture feature) were exposed to an altitude of 4,000 m above sea level, as simulated in a hypobaric chamber. Stepwise ascension was performed with a speed of 5 m/s starting at 450 m above sea level. A 5-minute rest was performed every 500 m to measure stimulation threshold at each step. After a stay of 30 minutes at 4,000 m stimulation threshold was measured, followed by a stepwise descent. Pacemaker interrogation and arterial blood gas analysis were performed at 450 and at 4,000 m, and a strength-duration curve was determined. Blood pressure, heart rate, and oxygen saturation were monitored continuously during the study. Ascent to 4,000 m above sea level induced a significant decrease in arterial pO2 (10.7 +/- 1.1 vs 5.5 +/- 0.3 kPa), pCO2 (5.3 +/- 0.3 vs 4.7 +/- 0.4 kPa), oxygen saturation measured by arterial blood gas analysis (95.5% +/- 1.2% vs 79.1% +/- 2.5%), and increase in pH (7.39 +/- 0.02 vs 7.45 +/- 0.04) (P < 0.0001). Stimulation thresholds and the strength-duration curve remained unchanged in all patients throughout the study. In conclusion, exposure to an altitude of 4,000 m above sea level with resultant hypobaric hypoxemia has no impact on ventricular stimulation thresholds. Therefore, in regard to the safety of pacing, pacemaker patients can safely be exposed to this altitude. PMID- 10793444 TI - Using intracardiac catheter recordings from the His and proximal coronary sinus to distinguish isthmus conduction block during catheter ablation of type I atrial flutter. AB - Isthmus conduction block, demonstrated with the use of multipolar catheter recordings, is considered the preferred endpoint for ablation of type I atrial flutter. This study investigated the feasibility of using recordings from the His and coronary sinus (CS) to document isthmus conduction block. Isthmus conduction block was produced with linear radiofrequency (RF) ablation in 27 patients with type I atrial flutter. In 13 patients (group I), RF was delivered until bidirectional isthmus conduction block was demonstrated with multipolar Halo catheter recordings. In 14 patients (group II), RF was delivered during pacing from the lateral isthmus at 600 ms until a reversal in activation of the proximal CS and His occurred. At this point, data from the Halo recordings were reviewed to see if reversal correlated with conduction block; if not, further ablation was performed until block was demonstrated. The initial reversal in His and CS activation during RF energy delivery correlated with isthmus block in only 4 (28.6%) of 14 patients in group II. Additional RF delivery produced isthmus block in the other ten patients resulting in a further increase in the St-CS interval of 35 +/- 20 ms. A His-CS interval of at least -40 ms signified isthmus block with a sensitivity and specificity of 48% and 100%, respectively. Reversal in His CS activation during pacing from the lateral margin of the isthmus is not specific for the creation of isthmus block. While activation of the proximal CS bipole > 40 ms after activation of the His appears specific for isthmus block, the low sensitivity of this finding limits its clinical use. PMID- 10793445 TI - Left bundle branch block with changing QRS morphology. PMID- 10793446 TI - The death of my father. PMID- 10793447 TI - Exercise induced atrioventricular block with gap phenomenon in atrioventricular conduction. AB - A 54-year-old man with normal atrioventricular (AV) conduction at rest gave a 4 year history of presyncope during exercise. Treadmill testing showed exercise induced AV block. Electrophysiological study demonstrated rate dependent infranodal AV block and abnormal refractory period of the His-Purkinje system. The gap phenomenon in AV conduction occurred during the programmed stimulation. Supernormal conduction could be considered as the mechanism of the gap phenomenon in this patient. PMID- 10793448 TI - His-Purkinje system reentry as a proarrhythmic effect of flecainide. AB - We report a case of tachycardia due to reentry within the His-Purkinje system (HPS) occurring after introduction of flecainide. The patient presented with a mild mitral regurgitation and normal left ventricular function. He had incomplete left bundle branch block with left-axis deviation. At the electrophysiology study, a prolonged HV interval was observed at baseline, and the tachycardia could be reproduced after ajmaline infusion. Six months after interruption of flecainide, the patient remains free of arrhythmia recurrence. The authors emphasize that proarrhythmic effects of flecainide may include reentry within the HPS in patients with underlying HPS disease. PMID- 10793450 TI - Thrombus formation at the site of radiofrequency catheter ablation. AB - A 55-year-old woman with a history of resected atrial myxoma with residual patched atrial septal defect (ASD) underwent a successful radiofrequency (RF) ablation of reentry atrial tachycardia. She presented with progressive dyspnea 5 days later. Transesophageal echocardiography revealed a 1-cm right atrial mass attached to the intraatrial septum at the ablation site. Repeat study after anticoagulation for 5 weeks showed complete resolution of the thrombus. Thrombus formation at the site of RF ablation is a potential complication that may require aggressive anticoagulation. Patients with patched ASD might be at higher risk. PMID- 10793449 TI - Ventricular tachycardia in a patient with primary hyperparathyroidism. AB - We present a case of primary hyperparathyroidism with hypercalcemia in a patient who had spontaneous attacks of ventricular tachycardia. Right ventricular burst pacing reproducibly induced ventricular tachycardia in the electrophysiological laboratory after intravenous administration of calcium-gluconate, and verapamil could terminate the tachycardia. After resection of the parathyroid adenoma, the calcium level was restored to normal, and ventricular tachycardia did not occur again during the follow-up period. PMID- 10793451 TI - Dissection of the coronary sinus secondary to pacemaker lead manipulation. AB - We report a 64-year-old man with atrial fibrillation and impaired left ventricular function in whom pacemaker lead manipulation during biatrial pacemaker lead removal and biventricular pacemaker lead insertion caused coronary sinus dissection. After further manipulation of the biventricular pacing lead, the true lumen of the coronary sinus could be entered and satisfactory left ventricular pacing undertaken. The dissection was managed conservatively without long-term sequelae. PMID- 10793453 TI - [An investigation on ethylene oxide sterilization management in medical institutions]. AB - The present study describes an investigation on the use and management of ethylene oxide (EO) which is used mainly as a sterilant in medical institutions in a Cabinet-order designated city. Information was obtained from a self administered questionnaire on safety and health matters related to sterilization with EO gas. The questionnaire was sent to 189 medical institutions including major hospitals and randomly selected clinics. 127 questionnaire were returned and the analysis was carried out for 120 respondents, excluding seven respondents whose answers were found to be inappropriate. The proportion of medical institutions, employing more than 50 workers, in which an occupational health physician and/or health supervisor was appointed was 70% and was lower than for other industries. 1.6% of the employees engaged in EO sterilization activities in the hospitals, whereas in clinics the percentage was as high as 20%. Several problems were found both in the management and work methods with EO: operations were not isolated in 46%; gas leaks and back-streaming of exhaust gas were found in 59% and 41%, respectively; and personal protective clothing was not used in 69%. Improvement was found only for three items among the institutions where an occupational health physician was appointed. These results suggest that good workplace controls and practices are essential for medical institutions using hazardous materials including EO for sterilization. PMID- 10793452 TI - Recommendations for extraction of chronically implanted transvenous pacing and defibrillator leads: indications, facilities, training. North American Society of Pacing and Electrophysiology Lead Extraction Conference Faculty. AB - The procedure of lead removal has recently matured into a definable, teachable art with its own specific tools and techniques. It is now time to recognize and formalize the practice of lead removal according to the current methods of medicine and the health care industry. In addition, since at this time the only prospective scientific study of lead extraction is the PLEXES trial, we suggest that studies relating to the techniques of and indications for lead extraction be designed. Recommendations for a common set of definitions, for a framework of training and reviewing physicians in the art, for general methods of reimbursement, and for consistency among clinical trials have been made. Implementation of these recommendations will require additional effort and cooperation from practicing physicians, medical societies, hospital administrations, and industry. PMID- 10793454 TI - [A study on effective implementation of the Shinjuku Regional Occupational Health Center Program in Japan]. AB - The Regional Occupational Health Center (ROHC) Program is aimed at improving occupational health services for small-scale enterprises (SSEs) with less than 50 employees. In 1993 the ROHC program began as a new policy announced by the national government, which entrusted the management to regional medical associations. The establishment of 347 ROHCs has completed in 1997. SSEs are able to use several services, such as workplace visits and health consultations at SSEs, but ROHCs have often been under-utilized. Purpose of this study was to discuss the effective implementation of ROHCs through a couple of surveys of potential and actual ROHCs users. Two surveys were conducted in the area of the Shinjuku ROHC. Survey 1: The subjects were 78 SSEs that used The Shinjuku ROHC between October 1996 and March 1999. A qualitative evaluation was carried out for 78 SSEs about users' positions, opportunities for use and kinds of consultations. Survey 2: The subjects were 50 users of the ROHC between April 1998 and March 1999. The effective response rate was 56.0%. A questionnaire survey was conducted on users' impressions before and after using the ROHC and on requests for further service. Survey 1 showed users' positions were office workers 36.5%, managers 32.0%, directors 17.0%, health nurses 9.0% and employees 3.5%, half of the users were recommend by the Labour Standards Inspection Office. The kind of consultation were on Industrial physicians, 22.9%, Occupational health control systems, 19.1% and ROHC functions, 16.8%. Survey 2 showed that almost but all users had misunderstood the ROHC's function before utilizing of its services, but many users were satisfied with the services provided. First, ROHCs need to make known their existence and the content of their services available to SSEs. Second, ROHCs have to improve their availability to SSEs. Third, ROHCs have to ensure the quality of their services to make users satisfied. And cooperation with other occupational health institutions will become important as the ROHC Program continues to develop. PMID- 10793455 TI - [A study on stress processes for college students during semester-end examinations]. AB - The present study examined individual differences in psychological and physiological stress responses to the same stressor. In Study 1, psychological stress responses were assessed at three time periods: four weeks before, the day before, and the middle day of the semester-end examination period (N = 69). Both stress coping and MPI were assessed once during the study. In Study 2, 15 subjects were exposed to the same psychological stressor in an experiment to measure heart rate. The stressors were memorization of material relevant to the examination (Test 1), and mental calculation irrelevant to the examination (Test 2). Subjects were divided into 2 groups on the basis of their Study 1 psychological stress response scores: the low stress group (LS, N = 10) and the high stress group (HS, N = 5). The results of Study 1 and Test 1 of Study 2 suggested that there were significant differences between LS and HS in stress responses. These findings were discussed in light of cognitive appraisal and controllability of time. Further studies are needed to clarify the aspect of time span in cognitive appraisal. PMID- 10793456 TI - [The time course of the oculogyral illusion and displacement of the subjective midline after self-rotation]. AB - We examined the time course of the apparent motion and displacement of the oculogyral illusion (OGI) after cessation of constant rotation (72 deg/s) as a vestibular stimulation. Subjects scaled the apparent motion of a target presented on the objective midline for 120 s after vestibular stimulation (Experiment 1) and the apparent displacement of the same target from the subjective midline (Experiment 2). The magnitude of apparent motion simply decreased from the maximum value as a function of time. In contrast, the magnitude of displacement was nearly zero, or localized near the subjective midline, immediately after the vestibular stimulation. Then, it increased rapidly in the direction of the acceleration, and decreased gradually moreover after 20 to 30 s. These findings suggest that the apparent motion and displacement in OGI could be controlled by different mechanisms, which have different response characteristics to the same vestibular stimulation. PMID- 10793457 TI - [Deficits of Morris water maze learning in rats with striatal kainic acid lesions]. AB - This study investigated effects of intrastriatal kainic acid administration, which induces selective neuronal cell death in the striatum sparing passing nerve fibers and terminals, on the acquisition of spatial learning in Morris water maze. Rats treated with bilateral kainic acid (0.5-1.0 microgram) into the striatum were trained to escape to a hidden platform under the water. Compared with control rats, kainic acid (1.0 microgram)-lesioned rats showed significantly longer escape latency in the place navigation, and in the following probe test lower rate of swimming within the quadrant where the platform had been placed. In addition, kainic acid (1.0 microgram)-lesioned rats showed longer latency in the cue navigation in which rats were trained to escape to a visible platform above the water. These deficits were caused by their tendency to swim along the wall of pool before approaching the platform and not by their swim difficulty. Results suggest that striatal neurons or neural circuits containing these neurons play an important role in the acquisition of spatial learning task, and the nature of this performance impairment was discussed in terms of both learning and attention deficits. PMID- 10793458 TI - [Attachment representation and a projective test with pictures of parent-child interaction]. AB - The purpose of this study was to assess individual differences in attachment representation. They were assessed, not through direct verbal reports, but indirectly as indicated in a projective test. The test required subjects to tell their impressions of pictures, which depicted daily, routine parent-child interactions. A series of pictures were developed for story-making task, which was named PARS (Picture Attachment Related Study). Three hundred and two (302) undergraduate and vocational students were asked to see the pictures, and freely imagine the situation, think what they would feel, and create the further story. They were then to recall their own experiences with their parents, and fill out a questionnaire of how they see their relationship with others. It was found that those who made a trustful PARS story recalled their own attachment experiences in an autonomous way, and had lower distrust in their relationship with others. Thus, results of the projective test were shown to reflect individual personal attachment experiences, and the test be useful. PMID- 10793459 TI - [Recovery of 3D structure from motion and its statistical efficiency]. AB - In order to investigate characteristics of the "structure from motion" in noise, we compared performances of recovering a three-dimensional structure from motion (3D condition) with those of recovering a two-dimensional structure (2D condition). In addition, we adopted a statistical efficiency approach to analyze the results. We conducted two experiments where we presented to observers a pair of three dots corresponding to three apexes of triangles rotating around an axis projected onto the fronto-parallel plane. A gaussian noise was added to the area of each triangle for each stimulus frame. Observers were required to discriminate areas of two triangles perceived with moving dots. Main results were as follows: (a) Significant difference was found between the efficiencies for 3D condition and 2D condition. We obtained about 50% or less recovery (i.e., a ratio of the 3D efficiency to the 2D one) in Experiment 2. (b) When we removed the constraints, the human efficiency and the percent recovery decreased dramatically. PMID- 10793460 TI - [Japanese anger expression styles and their interpersonal influence]. AB - The purpose of the present study was to investigate Japanese anger expression styles and their influence on interpersonal relationships. In the initial study, 239 undergraduates were asked to complete a questionnaire which assessed seven distinct anger expression styles. The results indicated that the participants most often employed three anger expression styles including hyojo-kucho (nonverbal), tohmawashi (implicit), and itsumodori (none). In the second study, 162 undergraduates were asked to watch these three anger expression styles portrayed on videotape. Participants rated the degree to which the actor in the videotape felt angry and their impressions toward male or female actors in either higher, lower, or equal status, same-sex situations. The findings indicated that the effect on interpersonal relationships varied across the three different expression styles. This study has important implications for understanding Japanese anger expression in interpersonal, and perhaps cross-cultural, relationships. PMID- 10793461 TI - [Perceived distributions of opinions in young generation and their parents' generation regarding sex roles]. AB - This study examined how people perceive the distributions of opinions about sex roles, in particular, how they saw generational differences in the opinions. Undergraduates and their parents were asked to estimate the opinion distributions in young generation as well as in their parents' generation. They were also asked to indicate their own opinions and the degree of their involvement with the issue. Main results were as follows: First, generation gaps were perceived; the respondents estimated that there would be stronger support for relatively liberal opinions in young generation than in their parents' generation, while the generation gaps were perceived in the reverse direction regarding relatively traditional opinions. Second, although this tendency was found for both sexes, it was more pronounced for female respondents. Third, fathers with higher personal involvement with the issue estimated stronger support for liberal opinions, while mothers with higher personal involvement estimated weaker support. This tendency in mothers was more salient in those who themselves supported relatively liberal opinions. PMID- 10793462 TI - [Effects of different explanations for performance on a learned helplessness task in undergraduates]. AB - Two groups of undergraduates were asked to solve anagrams and arithmetic problems. All the anagrams were solvable but part of the arithmetic problems was insolvable. After the preliminary task, one of the two explanations was given; internal attribution participants (Internal group, N = 22) were told that their failure was due to their problem-solving ability, while external attribution participants (External group, N = 21) were told that they failed because some of the problems had no solution. After the explanations, the groups worked on another set of solvable problems. The test task performance of internal group was significantly lower than that of External group (p = .05), and it was significantly lower than the group's preliminary task (p < .01). No deterioration effect was found for External group. These results supported the implications of the reformulated learned helplessness theory by Abramson, Seligman, and Teasdale (1978). In addition, relationships between performance and participants' attitude such as attributional style and optimism were examined with measurement before and after the task, but no significant correlation was found. PMID- 10793463 TI - Meningococcal disease falls in vaccine recipients. PMID- 10793464 TI - Serious hazards of transfusion (SHOT)--report for 1998-99. PMID- 10793465 TI - Exploring the potential for primary prevention. PMID- 10793466 TI - The privilege and responsibility of suicide prevention. AB - Each year about a million people worldwide take their lives, and a further unknown number, but probably no less than 20 million, attempt suicide. In addition, for every person who engages in suicidal behavior, another five or six will be associated with them in some way, making a conservative total of 100 million people worldwide who are affected each year--and to whom we have some degree of responsibility. There is no one approach to suicide prevention, and probably more so than for any other human condition, we are privileged to be able to collaborate with many different disciplines in our endeavors. However, there is a considerable responsibility to ensure that whatever our own area of expertise and interest may be, we should apply universal principals of objective analysis to these diverse contributions. This is addressed by examining research from four broad areas. First, there are studies that irrevocably bring together the sociological and biological approaches to suicide. Second, there are reports that support the notion of the universality of suicide. Third, despite considerable attention paid to the media, its influence on suicide is very limited. And finally, although there have been pessimistic reviews, there are persuasive data from innovative research designs that have documented that we can prevent suicide. PMID- 10793467 TI - Suicidal ideation and suicide attempts among Australian adults. AB - Too little is known about suicidal thoughts and behaviors at a population level to recommend appropriate preventive strategies. Using data from the Australian National Survey of Mental Health and Wellbeing (NSMHWB), this study examined rates of, and risk factors for, suicidal ideation and suicide attempts. For 10,641 respondents, the 12-month and lifetime cumulative incidence rates of suicidal ideation were 3.4% and 16.0%, respectively; the 12-month and lifetime cumulative incidence rates of suicide attempts were 0.4% and 3.6%, respectively. 12-month ideation was associated with anxiety disorders (relative risk ratio [RRR] = 3.51; population attributable risk percent [PAR%] = 19.6%), affective disorders (RRR = 11.94; PAR% = 38.8%) and substance abuse disorders (RRR = 1.85; PAR% = 6.1%). Attempts in the past 12 months were also associated with anxiety disorders (RRR = 7.06; PAR% = 37.0%), affective disorders (RRR = 12.24; PAR% = 39.5%), and substance abuse disorders (RRR = 2.09; PAR% = 7.7%). Age, marital status, and disability were associated with ideation; marital status and employment status with attempts. Approximately 12% of ideators in the sample progressed to making an attempt; employment status was the only significant predictor. Reducing suicidal ideation and attempts requires a multi-faceted approach that targets those with mental illness but also adopts population-based strategies that address other factors, such as unemployment. PMID- 10793468 TI - A study of suicides in Ballarat, Victoria, Australia. AB - OBJECTIVE: To study suicides occurring in Ballarat with regard to incidence, demographic variables, possible causal factors, and association with psychiatric disorders over a period of 5 years. METHOD: A detailed review of the coroner's record of every suicide occurring during 1992-1996 was undertaken. Information was obtained on socio-demographic variables, method and circumstances of suicide, and associated psychiatric disorders in each case and subjected to psychological autopsy. RESULTS: 75 suicides were recorded. The male to female ratio was 4:1 and average age was 43 years. 60% had associated psychiatric illnesses, mainly affective disorders. Carbon monoxide self-poisoning accounted for 40%, firearms for 30%, and hanging, overdose, asphyxia and other methods for the remaining 30%. It was statistically significant that the younger age group preferred firearms to other methods, and that their suicides were precipitated by interpersonal conflicts. Social and personal difficulties were associated in 33%, and triggering factors were present in 40%. Previous suicide attempts were present in 28%, while 32% had manifest behavior changes preceding suicides or verbalized their intent to suicide. CONCLUSIONS: Suicide rates in Ballarat were higher than the average overall Victorian and Australian rates. After a consistent decline over 4 years an increase occurred in 1996. The preferred method of suicide was carbon monoxide, although the young preferred firearms. Demographic and other psychosocial factors were similar to the rest of Australia. Unemployment was not a significant factor. Psychiatric conditions, personal and social problems figured prominently as factors of etiological significance in suicide subjects. PMID- 10793469 TI - The pattern of suicide in Pakistan. AB - There is paucity of information on suicide from Pakistan, an Islamic country in which data collection poses formidable challenges. A variety of social, legal, and religious factors make reporting and diagnosing suicide difficult. Paradoxically, incidents of suicide are regularly reported in newspapers in Pakistan. In the absence of other means these reports serve a useful, though basic information source for suicidal deaths. A 2-year analysis of all such reports in a major newspaper in Pakistan showed 306 suicides reported from 35 cities. Men (n = 208) outnumbered women by 2:1. While there were more single than married men, the trend was reversed in women. The majority of subjects were under 30 years of age and "domestic problems" was the most common reason stated. More than half the subjects used organophosphate insecticides, while psychotropics and analgesics were used infrequently. The study challenges the widely held belief that suicide is a rare phenomena in an Islamic country like Pakistan, and underscores the need for more culture specific research on this important public health problem. PMID- 10793470 TI - Child and adolescent suicide deaths in New Mexico, 1990-1994. AB - The suicide death rate in New Mexico is consistently higher than the national rate. Among adolescents, suicide is the third leading cause of death nationally, but in New Mexico it is the second leading cause of death. This study describes the pattern of adolescent suicide deaths in New Mexico. We conducted a retrospective review of all medical examiner autopsies for adolescent suicides (ages 20 years and younger) in New Mexico from 1990-1994. Records were reviewed for demographics and possible contributing factors such as depression, previous attempts, and alcohol and drug use. We identified 184 suicide deaths among children and adolescents ages 9-20 years for an overall rate of 12.9 per 100,000. Our rates for ages 5-9 years (0.2), 10-14 years (3.8), and 15-19 years (22.3) are over twice the U. S. rates. Suicide deaths resulted primarily from firearms (67%), hanging (16%), poisoning (6%), inhalation (4%), and other methods (7%). Method varied by ethnicity (p = .01) and gender (p = .03); males and non-Hispanic Whites were overrepresented among firearm deaths. Firearm ownership was known in 60 (48%) of the firearm deaths. Of these, 53% of the firearms belonged to a family member, 25% to the decedent, and 22% to a friend. Over one-third of decedents (41%) experienced mental disorders, primarily depressed mood and clinical depression. Previous suicide attempts were noted for 15% of the decedents. Some 50% of the decedents had alcohol or drugs present at the time of death; among American Indians/Alaska Natives, 74% had drugs or alcohol present (p = .003). Targeted interventions are needed to reduce adolescent suicide in New Mexico. We suggest raising awareness about acute and chronic contributing factors to suicide; training physicians to look for behavioral manifestations of depression; and involving physicians, teachers, and youth activity leaders in efforts to limit firearm accessibility, such as advising parents to remove firearms from their households. PMID- 10793471 TI - New approaches towards an understanding of deuterostome immunity. PMID- 10793472 TI - The antimicrobial host defense of Drosophila. PMID- 10793473 TI - Origin and evolution of the complement system. PMID- 10793474 TI - Genome paralogy: a new perspective on the organization and origin of the major histocompatibility complex. PMID- 10793476 TI - Recombination-activating genes, transposition, and the lymphoid-specific combinatorial immune system: a common evolutionary connection. PMID- 10793475 TI - Phylogeny of lower vertebrates and their immunological structures. PMID- 10793477 TI - Transcription factor expression in lymphocyte development: clues to the evolutionary origins of lymphoid cell lineages? PMID- 10793478 TI - The phylogenetic origin of antigen-specific receptors. PMID- 10793479 TI - Immunoglobulin isotypes: structure, function, and genetics. PMID- 10793480 TI - Evolution of vertebrate immunoglobulin variable gene segments. AB - Evolution of Ig V gene segments are generally characterized by (a) evolution by "the birth and death process" and (b) diversifying selection. However, the detailed evolutionary pattern of V gene segments varies among species due to the fact that the humoral immune system itself has changed during vertebrate evolution. The change in somatic diversification system coupled with the change in lymphocyte development has imposed a significant impact on the evolution of Ig genes. In order to understand the evolution of immunological genes it is important to view it in the context of the evolution of the entire immune system itself. PMID- 10793481 TI - The immune system of cartilaginous fish. PMID- 10793482 TI - Immune-type diversity in the absence of somatic rearrangement. AB - Immunoglobulin gene diversity has been characterized to varying degrees in modern representatives of all of the major radiations of cartilaginous fish. A pattern of overall chromosomal relationships of the various types of joined and unjoined Ig gene clusters is suggested in which the essential features are: (a) both Ig heavy and light-chain gene clusters occur on multiple chromosomes, (b) various classes of Ig are interspersed, (c) not all individual gene loci appear to be closely linked (Fig. 2). The cluster-type Ig gene system appears to be a series of (potentially) individually regulated loci analogous in part to the olfactory receptor gene system (BUCK and AXEL 1991) and markedly distinct from Ig loci in other vertebrate groups and TCR genes. Such a system would be ideal for the creation of variation in both form and function in a large number of clusters while preserving or partially preserving specificity in a number of other gene clusters. The full range of joined genes and the relative number of joined genes (as relates to unjoined genes), have yet to be determined. Nevertheless, a number of conclusions can be drawn: (a) four distinct forms of heavy-chain joining have been identified (VDD-J, VD-DJ, V-D-DJ, and VDJ; Fig. 1); (b) light-chain genes, which possess only two recombining elements, can be found in either unjoined (V J) or joined (VJ) forms (Fig. 1); (c) physical linkage between individual joined and unjoined genes has not been established, although such investigations have not been pursued in a significantly rigorous manner as to rule out this possibility; (d) joined light-chain genes are expressed and can be somatically mutated. Can germline joining be viewed as an ancestral character? The answer to this needs to be considered in the context of an overall system in which the level of structural and functional redundancy is extremely high. Joining is an adaptation that is unique to multicluster gene families. The phenomenon overcomes the possibility of not generating a specific form of a receptor, a major shortcoming of conventional rearranging Ig and TCR gene systems. The limitation of encoding specific receptors is compensated through large numbers of additional gene clusters that retain the capacity to rearrange and generate new specificities. Commitment of a V region to diverse, fixed specificity also is a property of the NITR genes, which although not related closely to Ig in a structural sense, may reflect an analogous phenomena. The possibility that immune type diversity is achieved in the absence of somatic rearrangement and that remnants of such systems could be operative in immune recognition in contemporary vertebrates is of extraordinary significance in terms of our overall understanding of the relationships between adaptive and innate immune recognition. PMID- 10793483 TI - Evolution and somatic diversification of immunoglobulin light chains. PMID- 10793484 TI - Evolution of the T cell receptor signal transduction units. PMID- 10793485 TI - A simplified procedure for rapid screening of vibrios for cholera toxin production. AB - Conventional methods for the detection of cholera toxin (CT) production by vibrios are not readily available to most laboratories. A modification is described here of a simplified method standardised earlier and based on the degradation of nicotinamide adenine dinucleotide (NAD) by CT; this is simple and can be carried out in small laboratories also. It is also easy to perform, and gives reproducible results. PMID- 10793486 TI - Isolation of recent variant influenza types A (H3N2) & B strains in Pune, India during 1998. AB - A total of 456 patients with influenza like illness were investigated during the course of continuous surveillance on influenza in Pune, India in 1998. The throat and nasal swab specimens collected from these patients were processed in MDCK cell culture. Influenza type A (H3N2) and type B strains were isolated in MDCK cell culture. They were identified as similar to the recently prevalent globally circulating variant strains; A/Sydney/05/97 (H3N2) and B/Harbin/07/94. PMID- 10793487 TI - Slime production as a virulence factor in Staphylococcus epidermidis isolated from bacterial keratitis. AB - A total of 126 coagulase negative staphylococci (CONS) isolated from corneal scrapings of patients of bacterial keratitis and 50 isolates from healthy eyes (controls) were tested for slime production. Eighty eight (69.84%) of 126 isolates from patients and 11 (22%) of 50 isolates from controls were slime producing (P < 0.001). Of these 88 isolates, 42 were Staphylococcus epidermidis biotype II, 30 were S. epidermidis biotype I, 8 were S. epidermidis biotype III and the rest belonged to CONS other than S. epidermidis. Amongst the corneal ulcer isolates, multidrug resistance (resistance to 3 or more antibiotics) was observed in 82.9 per cent (73/88) slime producing organisms as against only 18.4 per cent (7/38) nonslime producing organisms (P < 0.001). Similarly, of the total 99 slime positive and 77 slime negative isolates, 79 (79.8%) and 22 (28.6%) respectively were multidrug resistant (P < 0.001). Although, slime production is known to be one of the major virulence factors of CONS in extraocular systemic staphylococcal infections, the present study detected slime in isolates from ocular infections. It was found that S. epidermidis I and II were the common biotypes associated with bacterial keratitis and, slime production and multidrug resistance were the two important virulence factors. These observations have clinical and therapeutic significance. PMID- 10793488 TI - Isolation of emetine resistant clones of Entamoeba histolytica by petri dish agar method. AB - Emetine resistant clones of Entamoeba histolytica strain HM1:IMSS were isolated by using petri dish agar method after mutation with ethyl-methanesulphonate. Two emetine resistant clones were obtained and both were resistant to emetine at a concentration of 24 micrograms/ml of emetine. The 50 per cent inhibitory concentration (IC50) for both emetine sensitive and resistant clones was 5 and 14 micrograms/ml respectively. The colony forming efficiency of E. histolytica strain HM1:IMSS varied from 44 to 54 per cent. This method is useful for isolating clones from different strains of the parasite for molecular and immunological studies. PMID- 10793489 TI - Profile of neurologic disorders associated with HIV/AIDS from Bangalore, south India (1989-96). AB - One hundred patients (95 males, 5 females, mean age at presentation 31.6 +/- 9.4 yr) with various neurological disorders associated with HIV infection during 1989 1996 were evaluated at NIMHANS, Bangalore. Eighty patients belonged to group I associated with opportunistic neuroinfections and 20 to group II--non infectious neurological disorders. Cryptococcal meningitis either alone (n = 31) or associated with tuberculous meningitis (n = 6) was the most common (46.3%) followed by neurotuberculosis either alone (n = 24) or with cerebral toxoplasmosis (n = 4) accounting for 35 per cent. Other opportunistic neuroinfections included cerebral toxoplasmosis, herpes zoster, fulminant pyogenic meningitis and neurosyphilis. Clinical characteristics, diagnostic clues, their laboratory and radiological profiles and problems encountered in diagnosis and management of these opportunistic infections are highlighted. In group II (19 males and one female; mean age of 32.6 +/- 9.4 yr), two patients had cortical dementia, three acute brain stem involvement, two epilepsy and one had features suggestive of progressive multifocal leukoencephalopathy. Two patients of group I during follow up developed cortical dementia. Six had peripheral nervous system involvement similar to Guillain-Barre syndrome. Sixty six patients (63 of group I and 3 of group II) progressed to AIDS, 33 patients from group I and one patient from group II succumbed to the disease. With the rapid increase in the incidence of HIV/AIDS and an increase in the neurological manifestations of HIV/AIDS it is important to recognise the magnitude of the problem for health planning in India. PMID- 10793490 TI - A beta-thalassaemia allele with 3 base substitution in codons 4/5 & 6 (ACT CCT GAG-> ACA TCT TAG) detected by denaturing gradient gel electrophoresis & sequencing. AB - We report the analysis of a beta-thalassaemia gene involving three bases in codons 4/5 and 6 (ACT CCT GAG-> ACA TCT TAG) in a confirmed carrier whose child had beta-thalassaemia major. The fragment of the gene carrying the mutation was detected by denaturing gradient gel electrophoresis (DGGE) using GC clamped primers, followed by direct sequencing. DGGE analysis indicated that one gene was the wild type (normal) while the sequence changes observed were all in the other gene causing beta-thalassaemia major in the child. This confirms a single case report from Lucknow (UP) and adds to the beta-thalassaemia mutations identified in the beta-globin gene in India and will help in the thalassaemia control programme. PMID- 10793491 TI - Skeletal muscle endurance is reduced in chronically energy deficient adults. AB - Fifty eight adult males (well nourished; WN = 25, underweight; UW = 13, chronically energy deficient; CED = 20), aged 18 to 30 yr underwent an assessment of skeletal muscle function using a load cell based handgrip dynamometer coupled to a polygraph. WN subjects had higher handgrip strengths than either the UW or CED subjects (P < 0.05), though not when corrected for forearm muscle area or forearm volume. CED subjects fatigued faster than WN subjects during sustained maximal isometric contraction (P < 0.05) as well as during prolonged isotonic exercise (P < 0.05). During the latter, the onset of fatigue in the CED subjects was also faster than that in their anthropometrically similar UW controls. These data are consonant with reports of decreased productivity in undernourished adults during real life tasks. PMID- 10793492 TI - Long-term outcome for right heart function after biventricular repair of pulmonary atresia and intact ventricular septum. AB - OBJECTIVES: The right heart function of the patients with pulmonary atresia and intact ventricular septum was assessed periodically during the process of staged biventricular repair, and the implications for its long-term outcome were analyzed. SUBJECTS AND METHODS: During the period from 1971 to 1990, 21 neonates or infants with pulmonary atresia and intact ventricular septum had undergone initial palliative surgery. There were seven early postoperative deaths and one late death. Of the 13 survivors, 10 patients underwent subsequent biventricular repair and form the basis of this study. Their clinical records of roentgenography, electrocardiography, and catheterization studies at each staged period were reviewed retrospectively. RESULTS: Arrhythmia occurred late in 2 patients, one of whom died by arrhythmia at 11 years after definitive surgical repair. Therefore the actuarial survival rate was 85.7% at 14 years. The catheterization study after the definitive biventricular repair revealed a significant fall in the right heart pressure (p = 0.0005) and significant improvement in the right ventricular ejection fraction (p = 0.0004). In angiocardiography, dilatation of the right atrium was noted in all patients and was more marked in those who developed arrhythmia in conjunction with rapid growth of the right ventricle. Moreover, the serial repeated electrocardiography disclosed progressive and significant prolongation of both PQ interval (p = 0.003) and QRS duration (p = 0.021). CONCLUSIONS: Although biventricular repair for pulmonary atresia and intact ventricular septum proved to attain a satisfactory long-term result, it failed to resolve right heart dysfunction. Postoperative arrhythmia was prone to precipitate progressive dilatation of the right atrium. PMID- 10793493 TI - Autologous tissue-fragmented extracardiac conduit with rapid, stable endothelialization due to angiogenesis. AB - OBJECTIVES: Autologous angiogenic cytokines are known to activated by mincing stimulation, and well regulated in vivo. We applied this tissue fragmentation technique to a low-pressure pulmonary extracardiac conduit to obtain rapid endothelialization and stable neointima formation due to angiogenesis. METHODS: Subcutaneous adipose tissue was obtained, minced, suspended, and sieved through highly porous fabric vascular prosthesis by pressurized injection. The adipose tissue fragmented graft with an autologous fresh pericardial monocusp valve was implanted between the right ventricle and the pulmonary artery in 13 dogs. The same grafts without fragments were implanted in 8 dogs as controls. No anticoagulation therapy was given. RESULTS: Grafts were removed 6 to 1,128 days after implantation. In the developed grafts, angiogenesis occurred throughout the interstices of the graft wall from the adventitial side, and host cells proliferated and migrated. Endothelialization was completed throughout developed grafts at 2 weeks. The intima was still thin up to 1,128 days and free of degenerative changes. In control grafts, however, capillary infiltration was limited to perigraft tissue at 2 weeks and endothelialization was not completed by 3 months. Under the endothelial cell layer, laminal elastic fibers were formed through the developed graft wall by 4 months and still maintained at 1,128 days. CONCLUSIONS: The results demonstrated that adipose tissue fragmented extracardiac conduits induce rapid endothelialization and maintained thin intima with laminal elastic fibers. Long-term durability is expected based on results from using this technique in a low-pressure pulmonary system in dogs. PMID- 10793494 TI - Video-assisted thoracoscopic lobectomy reduces cytokine production more than conventional open lobectomy. AB - OBJECTIVE: We studied cytokine changes after video-assisted thoracoscopic lobectomy and conventional lobectomy in patients with stage IA lung cancer. METHODS: From June, 1997, 20 consecutive patients with stage IA non small-cell lung carcinoma underwent either conventional lobectomy via an open thoracotomy (n = 10) or video-assisted thoracoscopic lobectomy (n = 10). The cytokine concentration in serum and pleural fluid were measured for 6 days postoperatively. RESULTS: Interleukin-6 and interleukin-8 leads peaked at 3 h or 1 day after surgery. Cytokine levels in pleural fluid were more than 100 times higher than corresponding systemic levels. The increase of interleukin-6 in pleural fluid 3 hours after surgery was significantly smaller in video-assisted thoracoscopic lobectomy (3971 +/- 2793 pg/mL for video-assisted thoracoscopic lobectomy vs. 23274 +/- 8426 pg/mL for open lobectomy). There were no significant differences in the serum interleukin-6 and interleukin-8 concentrations between the 2 groups. CONCLUSION: The thoracoscopic approach lessened the increase of cytokines in pleural fluid, but benefits of reduced cytokine production in video assisted thoracoscopy remain to be clarified. PMID- 10793495 TI - Adoptive immunotherapy using specific cytotoxic T lymphocytes against human lung cancer-engrafted severe combined immunodeficiency mice. AB - OBJECTIVE: We studied the ability of human lung-cancer-specific cytotoxic T lymphocytes to suppress the growth of human lung adenocarcinoma (PC-9) engrafted in severe combined immunodeficiency mice. METHODS: PC-9-specific cytotoxic T lymphocytes were generated by multiple stimulation with irradiated PC-9 cells of regional lymph node lymphocytes from lung cancer patients expressing the same human leukocyte antigen-A locus haplotype as PC-9 following expansion due to the administration of immobilized anti cluster of differentiation 3 mAb and interleukin-2. Cytotoxic T lymphocytes showed specific cytotoxicity against PC-9 cells in vitro. Severe combined immunodeficiency mice with a subcutaneous graft of PC-9 were treated with a PC-9-specific cytotoxic T lymphocyte by i.v. injection and/or with interleukin-2 by s.c. injection. RESULTS: Cytotoxic T lymphocyte treatment suppressed PC-9 graft growth significantly an effect, significantly enhanced when combined with interleukin-2 injection. To evaluate the in vivo specificity of anti-PC-9 cytotoxic T lymphocytes, each mouse was subcutaneously inoculated in the right flank with PC-9, and in the left flank with A549 or Sq-1. Cytotoxic T lymphocytes plus interleukin-2 treatment was found to suppress PC-9 growth selectively, but not A549 or Sq-1 growth. CONCLUSIONS: These results provide sufficient rationale for conducting further clinical trials on immunotherapy using cytotoxic T lymphocyte for lung cancer patients. PMID- 10793496 TI - Surgical considerations of occlusive lesions associated with Takayasu's arteritis. AB - OBJECTIVE: Takayasu's arteritis is an inflammatory vascular disease of unknown etiology that affects the aorta and its main branches, requiring surgical intervention due to occlusive lesions. We studied early and late surgical results. METHODS: Between 1979 and December 1998, 46 patients--1 man and 45 women aged 15 to 72 years (mean: 46 +/- 13 years)--with occlusive lesions caused by Takayasu's arteritis underwent surgery. Preoperative steroids were administered to 22 patients having inflammation. Diagnosis indicating surgery included obstructive cervical vessel disease in 13 patients, obstructive coronary artery disease in 19, aortic coarctation in 15, and abdominal branch stenosis in 3. Surgical procedures involved bypass grafting in 31 (cervical vessel bypass in 13, ascending-abdominal aortic bypass in 4, axillary artery-abdominal aortic bypass in 10, descending-abdominal aortic bypass in 1, abdominal branch bypass in 3), coronary artery bypass grafting in 10, and coronary ostial endarterectomy in 9. RESULTS: Four (8.7%) died during hospitalization. Follow-up ranged from 1 to 240 months (mean: 117 months). Eight suffered late deaths and 6 patients died of cardiovascular problems. The total actuarial survival rate was 76.2% at 5 years and 70.5% at 10 years. CONCLUSIONS: Steroid therapy before and after surgery appears to affect the overall prognosis positively in patients with Takayasu's arteritis. PMID- 10793497 TI - Primary lymphoma of bone originating in a rib. AB - Malignant lymphoma originating in the bone is rare and is now recognized as being an independent clinicopathologic entity known as primary lymphoma of bone. A 60 year-old man complaining of right chest and back pain consulted our hospital for further examination. Chest X-ray and computed tomogram revealed osteolysis and a surrounding soft tissue mass in the sixth right rib. An ultrasonically-guided needle biopsy of the tumor was performed, and histologic examination indicated the dense proliferation of similar-sized atypical cells with nucleoli and an irregular nuclear border. A diagnosis of diffuse, medium-sized non-Hodgkin's lymphoma, B-cell type was made. En block resection of the tumor and chest wall was performed. Macroscopically, the tumor measured 7.5 x 4.8 x 3.0 cm in diameter, and the histologic findings were similar to those of the preoperative needle biopsy. Unfortunately, postoperative treatment with radiation therapy and chemotherapy was ultimately unsuccessful, and a local recurrence and metastatic lesions appeared in the stomach and para-aortic abdominal lymph nodes 7 months after the first symptom appeared. The patient died 3 months later. Surgery was chosen as the initial therapy as it was considered that a rib resection would not result in serious respiratory compromise and the complete resection of the tumor would be superior to radiation therapy for local control. Some authors have reported that the surgical resection of a primary lymphoma of the bone originating in a rib can yield a good prognosis. However, it is a systemic disease and a more effective therapeutic strategy should be developed. PMID- 10793498 TI - Aortoesophageal fistula caused by foreign body. AB - Aortoesophageal fistula is rare. A woman who developed aortoesophageal fistula after swallowing a fish bone developed hematemesis. 7 days later, we resected a false aneurysm near the left subclavian artery and repaired this section twice. Despite these measures, the woman died on hospital day 21. The clinical diagnosis was massive hematemesis from an infected aortic wall. The method of diagnosis, control of infection, and operative repair of aortoesophageal fistula are discussed. PMID- 10793499 TI - Profound hypothermia and low flow cardiopulmonary bypass in resectioning a massive facial arteriovenous malformation. AB - A 44-year-old woman underwent resection of a massive facial arteriovenous malformation under profound hypothermia and low-flow cardiopulmonary bypass. A left ventricular vent through a small left anterior thoracotomy avoided ventricular distention associated with peripheral cannulation. Low-dose aprotinin was used to improve hemostasis. These techniques thus show promise for the safe application of profound hypothermic cardiopulmonary bypass in noncardiovascular operations. PMID- 10793500 TI - Hemopneumothorax and hemoperitoneum in a case with large cell carcinoma of the lung. AB - Hemopneumothorax and hemoperitoneum coincide rarely in nontraumatic cases. Here, a 70-year-old male presented a left axillary lymph node and was diagnosed as having metastatic squamous cell carcinoma. Under the same diagnosis, another lesion developed in the right femur and was resected. One year later, computed tomography detected another tumor in the left adrenal gland. Shortly afterwards, left pneumothorax developed and a chest operation revealed hemopneumothorax due to a ruptured cavitary form of large cell carcinoma. The serum showed a human chorionic gonadotropin-beta level of 1,100 ng/ml. At three-months later, he died of hemoperitoneum. The autopsy demonstrated hepatic metastases and a ruptured adrenal metastasis; microscopy showed marked trophoblastic and squamous cell changes in these organs. This patient was unique in that the rupture of the pulmonary and the adrenal lesions caused clinical manifestation. PMID- 10793502 TI - The wisdom of a rural path. PMID- 10793503 TI - Physicians face challenges in the practice of rural medicine. PMID- 10793501 TI - Hilar lymph node metastasis in renal cell carcinoma. AB - A 48-year-old man, who underwent a right nephrectomy for renal cell carcinoma 7 years earlier, was found to have hilar lymph node metastasis alone, without lesions, in the pulmonary parenchyma. Chest X-ray and CT findings showed a left hilar mass about 4 x 2.5 cm in diameter. Left bronchial arteriography showed a hypervascular mass lesion in the left hilum. Macroscopic tumor invasion of the pulmonary artery and left main bronchus indicated left pneumonectomy. The resected specimen was found histologically to involve metastatic renal cell carcinoma in the left hilar lymph node about 3 cm in diameter. Tumor metastasis was limited to the lymph node. The metastatic pathway of renal cell carcinoma to the hilar lymph node was considered lymphogenous via either retrograde lymphatic flow from the thoracic duct or through the lymphatics in the inferior pulmonary ligament. PMID- 10793504 TI - Rural hospitals provide much-needed patient care. PMID- 10793506 TI - Preventive care in daily practice. AB - Advances in medical science have led to effective preventive steps that can be used by all physicians; however, most current preventive evaluations and applications are given in primary care settings. Hence, integrating preventive care into practice has become an important challenge for all primary care physicians. This article reviews the delivery of clinical prevention including implementation, barriers, and management. It is clear that preventive care leads to healthier people and that should be the goal of all medicine. PMID- 10793505 TI - Preventing carpal tunnel syndrome and cumulative trauma disorder: effect of carpal tunnel decompression excercises: an Oklahoma experience. AB - An Oklahoma meat packing company with 286 production workers experienced a high incidence of cumulative trauma disorder (CTD), specifically carpal tunnel syndrome (CTS). The company learned about the Carpal Tunnel Decompression Exercises (exercise program) through the American Academy of Orthopedic Surgeons' official web page. The Orthopaedic & Reconstructive Center assisted the company with implementation of the exercise program. The company reported their one-year follow up findings to the Governor's Council on Safety Policy (State of Oklahoma). The exercise program was the only ergonomic change the company implemented that year. The company's workers' compensation loss ratio improved from 13.89% to 11.61%. The company reported a net financial gain after comparing injury reduction versus work time lost to exercise. The incidence of Cumulative Trauma Disorder, excluding Carpal Tunnel Syndrome, was reduced 37% and Carpal Tunnel Syndrome was reduced 45.4%. The results of the exercise program were significant enough that the company made the exercise program permanent. PMID- 10793507 TI - State of the state's health. Oklahoma State Board of Health. PMID- 10793508 TI - State health agency promotes "Safe at Home" for Child Abuse Prevention Month. PMID- 10793509 TI - Utilization of routine medical services among immigrants from El Salvador. PMID- 10793510 TI - Improving recruitment and retention of medical scholarship recipients in rural Georgia. PMID- 10793511 TI - Achieving a credible health and safety approach to increasing seat belt use among African Americans. AB - African American youth are 60 percent less likely than children from other racial or ethnic backgrounds to be buckled up. Seat belt use among African American males has largely remained stagnant while that for other groups has increased. Overall, African Americans buckle up less often than other racial or ethnic populations. Seat belt use is a preventive health care action within public health. Clearly, a credible health and safety message is not being communicated effectively to African American communities. This paper proposes a number of possible solutions, including: recognition of the role that health care providers play in shaping patient or consumer attitudes and subsequent behavior in terms of prevention of disease and injury, educating physicians and health care providers to routinely recommend seat belt use especially for children, culturally appropriate educational safety programs, an improved relationship between law enforcement and communities, and zero tolerance for nonuse of seat belts. PMID- 10793512 TI - A single source of health care: does it affect health experiences for inner-city children? AB - This study was designed to determine whether having a single source of health care affects the health experiences of inner-city children. Seventy-five women were interviewed about the health experiences of their preschool-aged children (range 10-58 months) and were asked about utilization, emergency room use, preventive care, delays in care, and availability of a provider for the child's various medical needs. While most (94.7 percent) of the mothers identified a regular source of care for their child's preventive care, 74.7 percent named the same source for their child's sick care. There were no differences in most measures between mothers with a single source of care and those with multiple sources. However, those with a single source of care reported feeling their providers were more available (p < 0.05). Thus, naming a single source of health care may not be a useful indicator of better health experiences for inner-city children. PMID- 10793513 TI - Ethnic and racial differences in long-term survival from hospitalization for HIV infection. AB - This prospective cohort study compares 200 hospitalized, HIV-infected patients (Hispanic, African American, and white) from May 1992 to October 1998 to assess mortality (versus survival) over 75 months of follow-up. The relative risk of six year mortality for each ethnic group is compared using Cox proportional hazards models after controlling for sociodemographic and clinical characteristics, access to general medical care, and HIV-specific treatment. The median survival of Hispanics (15.5 months) was significantly (p < 0.05) shorter than that of whites (23.8); survival for African Americans (35.1) did not differ from whites. In multivariate analysis, the adjusted relative risk of six-year mortality for Hispanics compared with whites was 2.14 (95 percent confidence interval = 1.26 3.66). The poor outcomes of Hispanics was not explained by access to general care or by HIV-specific treatment. PMID- 10793514 TI - Health services utilization among Latinos and white non-Latinos: results from a national survey. AB - Utilization patterns may be changing as managed care organizations actively market services to Latinos. This study compares use of any care, emergency services, inpatient hospitalization, nonemergency outpatient care only, and preventive care among 1,001 self-identified Latino and 1,107 white non-Latino adults. Data were from the 1994 Commonwealth Fund Survey of Minority Health. Latinos were less likely than white non-Latinos to have entered the health system for any type of care, to have been admitted to a hospital, or to have used preventive care. Access to a regular source of care along with financial factors reduced the ethnic/racial gap in the use of any care and preventive care, yet cultural and behavioral factors contributed little. Latinos in managed care plans, compared with fee-for-service systems, were twice as likely to receive preventive care. This suggests that managed care has the potential to reduce inequities in preventive care utilization. PMID- 10793515 TI - Medicaid managed care in Chicago: changes in utilization. AB - In 1993, Illinois implemented Healthy Moms/Healthy Kids (HM/HK) in Chicago, a Medicaid managed care program for pregnant women and children. This study examines changes in immunizations for children (n = 134,072), prenatal care use for pregnant women (n = 5,151), and inpatient stays for mothers (n = 5,151) and newborns (n = 2,699) under the HM/HK program as compared with fee-for-service Medicaid in 1992 and 1993. HM/HK children were 10 percent more likely to receive any immunizations, and HM/HK pregnant women were 13 percent more likely to receive some prenatal care. Mothers' inpatient stays at delivery did not change under HM/HK. The length of newborn stays fell between 1992 and 1993, with both the HM/HK and the Medicaid 1993 comparison group deliveries associated with statistically shorter stays. During the early months of the program, improvements in the quantity of expected preventive care received were evident among children and women. PMID- 10793516 TI - Predicting health services utilization among homeless adults: a prospective analysis. AB - This study expands on the Andersen-Newman health services utilization (HSU) model. In a community-based homeless sample (n = 363) baseline predisposing, enabling, and needs-based variables predicted hospitalization and ambulatory outpatient service utilization within 1 year after baseline. Standard predisposing and enabling variables were supplemented with latent constructs representing substance use, mental illness, poor housing status, social support, community support, and barriers to health care. Need is represented by baseline health status. Poor physical health, more barriers, drug use, African American ethnicity, less community support, and less education predicted hospitalization, the least desirable form of HSU. Poor health, female gender, a regular source of care, community support, drug use, and fewer alcohol problems predicted an office visit. Because outpatient visits for acute conditions provide an opportunity for generally neglected preventive services and health screenings, this study suggests convenient multiservice health-related programs for the homeless that include drug and alcohol treatment. PMID- 10793517 TI - Medicare graduate medical education funding and rural hospitals. AB - To assess the importance of medical residents to rural hospitals, and to predict the possible effect of reductions in Medicare graduate medical education (GME) payments, data from Medicare hospital cost reports and from a telephone survey of rural hospitals with residency programs are analyzed. In prospective payment system year 11, 70 rural hospitals received more than $80 million in Medicare GME payments. The presence of rural training programs enhanced staff physician recruitment and retention and led to increased numbers of physicians settling in communities surrounding the facilities. Many survey respondents felt that elimination of GME funds would results in downsizing or outright elimination of their training programs. The results support the contention that rural training programs are important to hospitals and their surrounding communities and provide an essential component of the physician supply pipeline to rural areas. PMID- 10793518 TI - Mexican American infant mortality rate: implications for public policy. AB - Infants of Mexican American descent have lower infant mortality rates (IMRs) than do non-Hispanic blacks and non-Hispanic whites. Because IMR is used in allocation methods for primary health care resources, the result could be discrimination against Mexican American populations in the distribution of resources. This study examined the National Center for Health Statistics' infant birth and death records, as well as unpublished data from the Bureau of Primary Health Care. This study found that the low Mexican American IMRs are real and not simply a data anomaly and that inclusion of birth outcomes has a small and mixed effect on the designation of high-Hispanic areas as being medically underserved or short of primary health care professionals. The authors suggest inclusion of an additional high-Hispanic health indicator in the designation criteria for health resources. PMID- 10793519 TI - [Changes in brain metabolites under combined mild-hypothermia and isoflurane anesthesia in rat with temporal brain ischemia]. AB - The significance of mild-hypothermia as a treatment of brain ischemia-induced neuronal cell death was investigated by measurement of hippocampul excitatory amino acids, brain lactate and energy-charge just after 10 min of transient forebrain ischemia in a rat model with four vessels occluded. After 10 min of ischemia, cerebral circulation was restored. At that time, in the control group transient increases of hippocampul aspartate and glutamic acid levels were observed. Furthermore, brain lactate levels were also elevated but the energy charge was reduced. These significant changes were observed in the non-isoflurane anesthesia and mild-hypothermia rats (control group). However, in rats with treatment of either isoflurane or mild-hypothermia, the excessive amount of amino acids and the significant fluctuation of brain metabolic/energy pathway seen in the control group were suppressed. Particularly, in the combined treatment group, these increased and decreased phenomena induced by brain ischemia were significantly inhibited. In a group of pre- and post-treatment of mild hypothermia, the maximum peak of lactate was significantly less than that seen in the control group, although the sustained increased level of lactate was detected. These results indicate that the combined treatment with isoflurane anesthesia and mild-hypothermia is a suitable treatment for the brain dysfunction induced by ischemia and that the sustained hypothermia may help restore brain lactate levels after brain ischemia because of the lasting anaerobic metabolism. PMID- 10793520 TI - [Intrathecal 2% tetracaine causes mild histological lesions in the spinal cord without detectable sensory deficits on paw stimulation test in rats]. AB - The purpose of this study is to determine if intrathecal 2% tetracaine (TC) causes histological changes by its neurotoxicity, and to examine the relationship between the lesions and neurological functions. Twenty-two rats received either 2% TC or 0% TC dissolved in 10% glucose, via an intrathecal catheter terminated at T 13 level. Neurological deficits were evaluated by rat's behavior and paw stimulation test (UCSF). Five days after drug administration, the L 1 spinal cord with the anterior and posterior roots and cauda equina were excised for light and electron microscopy. Four rats out of 8 in 2% TC group showed mild pathological changes induced by neurotoxicity mainly in the posterior roots and slightly in the posterior column. However, there were no significant differences in sensory and behavioral function between the rats who had received 2% TC with lesion and the others with no lesion. As many rootlets enter one segment of the spinal cord, mild and restricted lesions may be difficult to detect by sensory tests. These findings may explain the fact that the patients with transient neurologic symptoms (TNS) are normal by neurological tests. PMID- 10793521 TI - [Effects of 4-hydroxybenzoic acid methyl ester on the Ca(2+)-related functions of skinned skeletal muscle fibers from the guinea pig]. AB - The amide-linked local anesthetics can be used safely in patients susceptible to malignant hyperthermia (MH). 4-Hydroxybenzoic acid methyl ester contained as a preservative in local anesthetic solution was suspected to have triggered MH in a patient who had received nerve block with lidocaine solution. The author investigated the effects of 4-hydroxybenzoic acid methyl ester on Ca(2+)-related functions of the skeletal muscle using saponin-treated skinned fibers from the extensor digitorum longus muscle of guinea pigs. 4-Hydroxybenzoic acid methyl ester accelerated the Ca2+ induced Ca2+ release (CICR) rate from the sarcoplasmic reticulum (SR) in a dose-dependent manner, and the Ca2+ concentration-CICR rate curves were shifted to the left with 4-hydroxybenzoic acid methyl ester at concentrations above 30 microM. 30 microM of 4-hydroxybenzoic acid methyl ester is associated with a plasma concentration of lidocaine intoxication which induces general convulsion. Ca2+ uptake, initial rate of Ca2+ uptake by the SR and Ca2+ sensitivity of the contractile system were not affected by 100 microM of 4 hydroxybenzoic acid methyl ester. These results suggest that amide-type local anesthetic solutions and agents containing 4-hydroxybenzoic acid methyl ester as a preservative may trigger MH in MH susceptible patients, if they are accidentally administered into the blood vessel. PMID- 10793522 TI - [Continuous monitoring of hemoglobin oxygen saturation of the hepatic vein in patients undergoing laparoscopic cholecystectomy]. AB - We monitored of hemoglobin oxygen saturation of the hepatic vein (ShvO2) continuously in 6 patients undergoing laparoscopic cholecystectomy under inhalation anesthesia only, and in 6 patients under combined epidural and inhalation anesthesia. In inhalation anesthesia only group, arterial blood pressure increased and ShvO2 values decreased immediately after intraperitoneal insufflation with CO2, demonstrating the mean lowest ShvO2 value of 25 percent. In combined anesthesia group, arterial blood pressure and ShvO2 values were unchanged after the intraperitoneal insufflation compared with values obtained before the insufflation. The results indicate that hepatic blood flow may be better preserved with combined epidural and inhalation anesthesia than with inhalation anesthesia only, suggesting that sympathetic block by epidural anesthesia may contribute to this favorable effect on ShvO2. PMID- 10793523 TI - [A review of correlation between transfusion rate of irradiated blood and potassium load]. AB - Transfusion of the irradiated blood has become popular in Japan to prevent the graft-versus-host disease. Case reports, however, of hyperkalemic death or lethal ECG change have been increasing since the irradiated blood was introduced nation wide. The potassium concentration of the supernatant of irradiated blood becomes about 60 mEq.l-1 by the end of storage period. If the potassium concentration is 60 mEq.l-1, the estimated safe transfusion rate would be 6 ml.min-1 and this can not be agreed with by clinicians who transfuse daily in cases of massive bleeding. The calculated safe transfusion rate (10 mEq.hr-1 of potassium load) ranges from 6 to 72 ml.min-1 considering the storage period from the day of gathering and irradiation. This difference could affect the survival rate. On the other hand, only the difference of the potassium concentration in the supernatant between the irradiated blood and the non-irradiated blood could not explain the increasing number of hyperkalemic case report. The mechanical factors may be related to hemolysis that causes extreme hyperkalemia. Control studies of proper transfusion rate of irradiated blood should be performed to establish a safety guideline for rapid transfusion of irradiated blood. PMID- 10793524 TI - [Two cases of extreme hemodilution caused by massive hemorrhage immediately after start of operation]. AB - We describe two cases of extreme hemodilution due to large amounts of fluid infusion for unexpected massive hemorrhage. In both cases, unexpected hemorrhage with difficult hemostasis occurred within 60 min after the start of the operation. For lack of transfused blood, large amounts of fluid infusion using crystalloid and colloid solutions including 5% albumin, plasma expander and lactated Ringer's solution were administered to maintain circulatory blood volume. The hemoglobin concentration and hematocrit had been below 2.0 g.dl-1 and 10% for approximately one hour, respectively. The extreme hemodilution improved by the urgent blood transfusion. In one case, intraoperative autotransfusion with Cell-Saver was performed. In spite of intraoperative extreme hemodilution, their postoperative courses were uneventful. Intraoperative awareness was present in both cases. PMID- 10793525 TI - [Studies on surfactant replacement therapy in pediatric measles pneumonitis]. AB - Measles pneumonitis, as well as encephalitis, is the most important complication associated with mortality in measles. Many medications including steroids and vitamin A have been applied to pediatric measles pneumonitis. However, the efficacy of such medication has not yet been established. This study is aimed at evaluating the effectiveness of surfactant replacement therapy in pediatric measles pneumonitis. Five patients (aged 1-2 years) with measles pneumonitis were transferred to our emergency center. On the transferred day, Surfactant-TA was administered by intratracheal method. After administration of surfactant, PaO2/FIO2 increased from 63.6 +/- 11.0 (mean +/- SE) to 206.2 +/- 54.1 in an hour and to 163.8 +/- 34.8 in 24. At the same time, the CO2 elimination and the dynamic compliance were improved. Because of these effects, the peak inspiratory pressure employed in mechanical ventilation could be reduced. It is concluded that surfactant replacement therapy can prevent the patients with measles pneumonitis from hypoxemia and ventilation-induced lung injury. However, further study is needed to maintain the improved oxygenation. Recently, it is reported that the effect of exogenous surfactant on oxygenation and activity of pulmonary neutrophils is regulated by the amount and/or concentration of administered surfactant. Therefore, it is an urgent issue to find out the optimum amount and concentration of exogenous surfactant used clinically. PMID- 10793526 TI - [Preliminary report: first identification of known mutation in the ryanodine receptor gene in a Japanese malignant hyperthermia pedigree]. AB - In Japan, the rate of Ca-induced Ca release (CICR) using skinned fibers of skeletal muscle has been employed as a diagnostic test for malignant hyperthermia (MH) susceptibility, since most of the typical fulminant MH patients showed an enhancement of CICR rate. Recently the Ca releasing channel responsible for the CICR was reported to be mainly the ryanodine binding Ca protein which was identified as the causative site of MH muscle according to recent genetic studies. Among patients with enhanced CICR rate, one point mutation for Arg 2434 His was recognized in a family. Although this site is related to the central core disease (CCD), this pathological change could not be observed in this patient. This is the first report describing a gene mutation in a MH family identified in Japan. PMID- 10793527 TI - [Anesthesia for MIDCAB (minimally invasive direct coronary bypass) in a patient with Tangier disease: a case report]. AB - Tangier disease is a rare, autosomally-inherited disorder of lipoprotein metabolism characterized by absence or marked deficiency of normal high density lipoprotein (HDL) cholesterol in plasma resulting in the accumulation of cholesteryl esters in various organs. The patient was a 55-yr-old male diagnosed as Tangier disease 16 years before. He had angina on exercise and his coronary angiogram revealed triple vessel disease including left main trunk (LMT) lesion. Stenosis of the right coronary artery was treated by percutaneous transluminal coronary angioplasty (PTCA). He was scheduled for a MIDCAB for further PTCA to be performed to relieve the stenosis of LMT. Preoperative laboratory data and physical examination showed total cholesterol 36 mg.dl-1, HDL-cholesterol 2 mg.dl 1, apoprotein A-I not-detected, pancytopenia, hyperplastic orange tonsils, splenomegaly and hepatomegaly. Clonidine 0.225 mg was orally given as a preanesthetic medication. Anesthesia was induced with fentanyl and midazolam and maintained with propofol, sevoflurane and supplemental fentanyl. Nitroglycerin and diltiazem were infused continuously. ST segment was elevated transiently during the clamping of the left anterior descending branch. Hemodynamic parameters were stable during the operation. He was extubated 2 hours after the end of the operation. No significant changes were found in postoperative EKG, total cholesterol, HDL-cholesterol and triglyceride. Perioperative course was uneventful. PMID- 10793528 TI - [Use of intubating laryngeal mask airway for three patients with difficult intubation]. AB - We used intubating laryngeal mask airway (ILM) for three patients with difficult intubation, and tracheal intubation was successfully performed through the ILM in all three cases. Difficult intubation in the first case was caused by direct invasion of malignant lymphoma into the right maxillary sinus leading to restricted mouth opening. Neck stiffness due to invasion of metastatic cancer into the cervical spine in the second case and facial trauma caused by traffic accident in the third case gave rise to the difficult intubation. Insertion of the ILM was successfully performed in all the patients following induction of general anesthesia, and the ventilation through the ILM was possible without any difficulties. Subsequently, all patients were intubated through the ILM successfully. We realized that the ILM is useful and should be prepared on the induction of anesthesia in patients suspected of difficult intubation. PMID- 10793529 TI - [Anesthetic management of a patient with hypohidrotic ectodermal dysplasia]. AB - We anesthetized a 10-year-old girl with hypohidrotic ectodermal dysplasia for an ophthalmic surgery. Ectodermal dysplasia involves the abnormalities of ectodermal tissues and has a triad; hypohidrosis, a lack of teeth, and the scarcity of hair. Hyperthermia may occur due to the defect of sweat glands. Therefore, the body temperature must be monitored continuously. Respiratory tract infection occurs frequently due to the absence of seromucosal glands. We recommend humidifying the inspired gases during the operation. Tracheal intubation may be difficult because of maxillary and/or mandibular abnormalities. We conclude that the particular care should be taken such as the management of the body temperature, preparation for the difficult airway and the humidification of respiratory tract. PMID- 10793531 TI - [Arytenoid dislocation caused by endotracheal intubation]. AB - We experienced arytenoid dislocation in a 17-year-old man with no significant medical history, who underwent anterior cruciate ligament reconstruction under general anesthesia. After induction of anesthesia, the trachea was easily intubated with an 8.0 mm cuffed endotracheal tube. The operation lasted approximately four hours. The patient was extubated uneventfully. On the first operative day, the patient complained of hoarseness. Four weeks had passed before the diagnosis of right arytenoid cartilage dislocation was made by fiberoptic laryngoscopy. Voice therapy was started. Three weeks after starting voice therapy, the patient's voice was restored almost to the preoperative status. PMID- 10793530 TI - [Anesthetic management for a patient with autoimmune hemolytic anemia]. AB - A 69-year-old woman with autoimmune hemolytic anemia (AIHA) received gastric resection and splenectomy under general anesthesia combined with epidural anesthesia. Her blood type had been determined as type A, Rh type CCDee, with anti-e-antibody and anti-pdl-antibody positive as autoantibodies. We applied the technique of hemodilutional autologous transfusion to supplement the blood loss during the operation to prevent hemolysis or occurrence of new antibody after homologous transfusion. We should pay much attention to a patient with AIHA to find signs of peri-operative hemolysis and to give the treatment for hemolysis in early stage. We consider that the hemodilutional autologous transfusion is a useful technique for the anesthetic management of the patient having a rare blood type. PMID- 10793532 TI - [Investigations on the position and safety of Swan-Ganz catheters]. AB - Swan-Ganz catheters (S-G catheter) may cause many complications. We measured floating distance (FD) of catheter tip with balloon inflation or deflation, and distal movement (DM) before and after the operation. FD and DM were significantly altered with 7 Fr. S-G catheters. These results suggest that careful observation is required in order to prevent unpredictable pulmonary artery perforation and infarction during the use of 7 Fr. S-G catheters. PMID- 10793533 TI - [A patient for craniotomy with ECG abnormality occurring on admission to the operating room]. AB - We report a 72 year old male patient for craniotomy with ECG abnormality occurring on admission to the operating room. His preoperative ECG showed normal sinus rhythm without ST-T change and T wave abnormality. On admission to the operating room, negative T wave was noted on the ECG monitor. Therefore, we applied isosorbide dinitrate tape after the induction of anesthesia. When the dura mater was opened, subarachnoid hemorrhage was noticed. Intraoperative pathological examination revealed a glioblastoma multiforme. The tumor bled from inside and outside. The perioperative T wave abnormality was probably due to subarachnoid hemorrhage or bleeding from the tumor itself. The T wave abnormality disappeared on the 10th postoperative day. PMID- 10793535 TI - [Report of Sata Clinical Fellowship; brain death and organ donation in hospital for sick children, Toronto, Canada]. AB - I have experienced two cases of pediatric organ donation from the brain dead patients in Hospital for Sick Children in three months. First case was a 9-year old boy after a traffic accident. Second case was an 11-year-old boy with intracranial hemorrhage. Brain death is diagnosed by clinical criteria alone in Canada, as in many of developed countries. EEG or brain flow studies are not mandatory. In the first case, brain death was confirmed after additional brain flow study, EEG, and SSEP because of cervical spinal injury. Second case was diagnosed as brain death by clinical criteria alone, and cardiopulmonary resuscitation was performed after brain death diagnosis. MORE (multiple organ receival and exchange program of Ontario), Organ Donation Team (critical care physicians, nurses, organ donation coordinators, social workers and chaplains) in HSC, and volunteers play the important role to help the family and to make the organ transplantation successful. In Canada, pediatric brain death and organ donation are widely accepted, but there remains an imbalance between the demand for transplantation and the number of organs available. PMID- 10793534 TI - [Hypercapnea after minitracheotomy]. AB - Minitracheotomy is a useful procedure for the respiratory management of patients with sputum retention observed frequently in chronic lung disease and postoperative respiratory failure. However, we experienced a case of hypercapnea after minitracheotomy. An 83-year-old woman was scheduled for gastrectomy. She had respiratory failure of Hugh-Jones 5 with fibroid lungs and the infection with atypical Mycobacterium and MRSA. Her sputum was adhesive and showed persistent increase during pre- and intra-operative periods. After the operation, minitracheotomy was carried out against the sputum retention by using Portex Mini Trach II kit. Hypercapnea, however, was observed in spite of no evidence of the surgical complications such as bleeding or inadequate care for sputum from the inserted tube. Bronchofiberscope revealed adhesive sputum stuck to the space between the inserted tube and the trachea. In this case, hypercapnea was improved by opening the top of the inserted tube and frequent care for sputum. In cases with a large amount of adhesive sputum, sputum retention may easily occur not only at the peripheral bronchial branch but also between the inserted tube and the trachea and it can cause hypercapnea which may be overlooked. PMID- 10793537 TI - [Topics in the cataract research]. PMID- 10793536 TI - [Electricity generators for disastrous emergencies]. PMID- 10793538 TI - [Identification of a free non-tryptophan fluorophore in water-soluble fraction of human brunescent cataractous lens nucleus]. AB - PURPOSE: We previously reported that a unique free fluorophore (Fl-Glc), presumably a beta-glucoside, is particularly abundant in human brunescent cataractous lens nuclei. Our preliminary experiments indicated that incubation of low-molecular weight (MW) fraction of non-brunescent lens nuclei causes an increase in a particular fluorophore (Fl-X). This study was undertaken to compare the Fl-Glc with the Fl-X and subsequently to identify the Fl-X. METHODS: Experiment 1. The purified Fl-X and its beta-glucosidase digest (aglycon) were compared with the Fl-Glc and its aglycon, respectively, by high-performance liquid chromatography (HPLC). Experiment 2. i) The Fl-X and its aglycon were analysed by liquid chromatography/mass spectrometry (LC/MS). ii) Authentic xanthurenic acid was analysed by HPLC and LC/MS. RESULTS: Experiment 1. The retention times of the Fl-X and the Fl-Glc exactly coincided. The fluorescence peaks of both disappeared after beta-glucosidase treatment. Experiment 2. i) LC/MS results suggested that the MWs of the Fl-X and its aglycon were 367 and 205, respectively. ii) HPLC and LC/MS results for xanthurenic acid (MW = 205) were exactly the same as those for the aglycon of the Fl-X. CONCLUSIONS: The Fl Glc and the Fl-X are identical, and the Fl-X (= Fl-Glc) is a glucoside of xanthurenic acid. PMID- 10793539 TI - [The effect of intraocular lidocaine in white rabbit eyes]. AB - PURPOSE: Recently, intraocular lidocaine anesthesia has been used in cataract surgery. We studied the toxicity of intraocular unpreserved lidocaine for corneal endothelial cell and retina using Japanese white rabbits. METHOD: They were divided into two groups. One group was injected intracamerally and the other group was injected intravitreally with 0.2 ml of unpreserved lidocaine of 0%, 0.02%, 0.2%, or 2% concentration. The number of corneal endothelial cells was measured 1 week after the injection. The rabbits were killed after measurements, and their corneas were studied histologically. The retina was examined by electroretinogram from before the injection through 1 week after the injection. RESULTS: There was no significant change in number of corneal endothelial cells after injection of 0.2% lidocaine. However, histological abnormality was seen in corneal endothelial cells after 2% lidocaine injection. There was also significant change in electroretinogram with 2% lidocaine injection. No histological abnormality was seen in the retina 1 week after the injection. CONCLUSION: The rabbit cornea and retina manifested no serious changes after the injection of lidocaine at less than 0.2% concentration functionally and histologically. PMID- 10793540 TI - [Immunohistochemical study of apoptosis of lens epithelial cells in human and diabetic rat cataracts]. AB - PURPOSE: To evaluate apoptosis of lens epithelial cells with immunohistochemical methods. METHODS: We performed terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays on capsulotomy specimens (68 eyes in 53 patients) from patients who had undergone cataract surgery and an epithelium of diabetic cataracts in rats (144 eyes in 72 rats). The animal model of diabetic cataracts was prepared by injection of streptozotocin in three-week old rats. The rats were also examined using the proliferating cell nuclear antigen (PCNA) immunohistochemical staining method. RESULTS: Although some TUNEL-positive cells were detected in capsulotomy specimens, we recognized little correlation between its distribution and morphological classification of cataracts. In the animal model of diabetic cataracts, TUNEL-positive cells were seen around the region where epithelial cells had accumulated. In the accumulated region, PCNA labeled cells undergoing DNA synthesis were also detected. CONCLUSION: These results suggest the possibility that apoptosis occurs in human lens epithelial cells and apoptosis and proliferation may be induced by factors such as hyperglycemia. PMID- 10793541 TI - [Effect of carteolol hydrochloride on ocular blood flow dynamics in normal human eyes]. AB - PURPOSE: The effect of topical 2% calteolol hydrochloride on the ocular blood flow dynamics in normal human eyes was studied. MATERIALS AND METHODS: Ten healthy volunteers were studied using color Doppler imaging and scanning laser Doppler flowmetry. In the first experiments, one eye received 30 microliters of 2% carteolol hydrochloride twice daily for 7 days and the fellow eye a placebo in a randomized masked manner. The blood flow velocity of the central retinal artery, the ophthalmic artery, and the short posterior ciliary artery, the blood flow volume of the peripapillary retina (PPR), intraocular pressure (IOP), ocular perfusion pressure (OPP) and pulse rate (PR) were measured before treatment, 2 hours after the initial instillation, and after the last instillation on the 7th day. In the control experiments, 30 microliters of placebo was instillated in both eyes, and the above parameters were measured according to the same time schedule as in the first experiments. RESULTS: In the carteolol-treated eyes, the systolic maximum (p = 0.012) and diastolic minimum (p = 0.019) blood flow velocities of the ophthalmic artery, and the blood flow volume of the superior (p = 0.003), central (p = 0.001) and inferior (p = 0.000) PPR showed significant increases in the first experiments compared with the control experiments. IOP and OPP in both eyes showed significant reduction in the first experiments compared with the control study. CONCLUSION: These results indicated that topical carteolol hydrochroride increases the blood flow of the intraorbital microcirculation and PPR. PMID- 10793542 TI - [The Rayleigh color matches in idiopathic macular holes treated by vitrectomy]. AB - PURPOSE: The Rayleigh color matches were measured to investigate the optical density in the cone photopigment of the central retina in patients with macular holes that had closed after vitrectomy. METHODS: The Rayleigh equation was measured with the IF-2 anomaloscope in 7 patients with macular holes in one eye, that had closed after vitrectomy and gained an improvement in visual acuity of 0.5 or better. Reductions in the pigment optical density of cone photoreceptors of the opposite eyes were estimated with Rayleigh equations obtained in this experiment. RESULT: The Rayleigh equations of the affected eyes were shifted toward protanomalous setting compared with that of the opposite eyes. The optical densities of cone photoreceptors of the affected eyes were lower than those in the opposite eyes, even in patients with visual acuity of 1.0 or better. CONCLUSION: These findings suggest that the recovery of the visual acuity may precede that of the optical density in cone photopigment of central retina. The Rayleigh equation is more sensitive than visual acuity for evaluating visual function after vitrectomy for macular holes. PMID- 10793543 TI - [Vitrectomy for the treatment of expulsive hemorrhage]. AB - PURPOSE: To evaluate the surgical outcome of vitrectomy in the treatment of expulsive hemorrhage associated with intraocular surgery. METHODS: We reviewed 12 eyes from 12 patients with expulsive hemorrhage, occurring after or during cataract extraction (4 eyes), phacoemulsification (2 eyes), glaucoma filtering surgery (4 eyes), or vitrectomy (2 eyes). Mean follow-up period was 21 months. RESULTS: The retina was reattached in 6 eyes (50%) after the initial surgery and ultimately in 9 eyes (75%). Three eyes, which failed to achieve retinal reattachment, resulted in phthisis bulbi. Final visual acuity was 0.1 or belter in 4 eyes and 0.01 to 0.09 in 4 eyes. The incidence of expulsive hemorrhage was 0% for cataract surgery, 0.57% for trabeculectomy, and 0.09% for vitrectomy at Toho University Sakura Hospital. CONCLUSION: In the treatment of expulsive hemorrhage, vitrectomy is an effective surgical procedure to improve the visual function. PMID- 10793544 TI - [The difference in saccadic parameters among several visually guided tasks- saccadic reaction time]. AB - PURPOSE: The saccadic reaction time (SRT) was measured as a saccadic movement parameter using visually guided tasks. SUBJECTS AND METHOD: The SRT was measured for nine healthy adult subjects aged from 28 to 49 years. The visually guided tasks for saccadic movement induction were simultaneous, gap, overlap, delayed memory, and anti-SM tasks. RESULTS: The median of the SRT (msec) for each task in the actual measurements was as follows: 216 for the simultaneous task, 180 for the gap task, 240 for the overlap task, 234 for the delayed-memory task, and 292 for the anti-SM task. The SRT distribution of each subject exhibited two peaks for the gap task but mostly one peak for each of the other tasks. The first peak, observed in the SRT distribution for the gap task was an eysress saccade. CONCLUSION: The SRT for the gap task was the shortest and the SRT got longer in the order of the simultaneous task, and the overlap task, and the anti-SM task. Some subjects easily showed an express saccade in the gap task, but others did not. PMID- 10793545 TI - [Multifocal electroretinogram in central serous chorioretinopathy]. AB - PURPOSE: To assess retinal function topographically in the affected eyes with central serous chorioretinopathy (CSC). PARTICIPANTS AND METHOD: We recorded multifocal electroretimogram (mERG) in 5 patients with unilateral CSC, and analyzed the topographical properties in the central visual field (rings #1 + 2). RESULTS: mERG amplitudes in the CSC eyes were significantly reduced compared with the fellow eyes, for P 1-N 1 (58.5 +/- 23.5%, p < 0.05) and for P 1-N 2 (47.5 +/- 15.0%, p < 0.05). mERG latencies in the CSC eyes were significantly increased compared with those in the fellow eyes, for N 1 (by 9.0 +/- 8.1%, p < 0.05) and for P 1 (by 8.4 +/- 7.0%, p < 0.05). mERGs in the CSC eyes gradually recovered in the follow-up period. However, the responses did not recover to normal value during follow-up, even when the subretinal fluid disappeared ophthalmoscopically. CONCLUSIONS: These results show that a topographical analysis of the mERG is useful for clinical observation of CSC. PMID- 10793546 TI - [A case of reversed ophthalmic artery flow without occlusion of the internal carotid artery]. AB - BACKGROUND: We report a case of reversed ophthalmic artery (OA) flow showing neither occlusion of the internal carotid artery (ICA) nor ophthalmic symptoms. CASE: An 82-year-old man had transient blindness in both eyes. After left carotid endarterectomy, ophthalmic symptoms improved in the right and left eyes. Afterwards, he had a relapse in the left eye. RESULTS: There was no unusual impression in the ophthalmic examinations, but color Doppler imaging showed reduced flow in the left OA, reversal of flow direction in the right OA. The central retinal artery (CRA) and short posterior ciliary arteries (SPCA) were not detected in the left eye but were detected in the right eye. Digital subtraction angiography demonstrated that the right OA was not contrasted with the right ICA, despite there being no stenosis or occlusion of the right ICA. CONCLUSIONS: We suggest that carotid surgery may affect the ocular circulation of the opposite side, and reversed OA flow as a collateral pathway from the external carotid artery may occur in the absence of carotid artery stenosis. Good circulation of the CRA and SPCA may preclude manifestation of ophthalmic symptoms even if the OA flow is reversed. PMID- 10793547 TI - [Twenty-three cases of primary cornea guttata]. AB - PURPOSE: To evaluate the clinical aspects of patients with primary cornea guttata and the morphology of their corneal endothelial cells. METHODS: Twenty-three patients who visited Kanazawa Medical University Hospital or related hospitals in Ishikawa or Fukui prefectures and were diagnosed as having primary cornea guttata by slit lamp microscopy and contact specular microscopy underwent an analysis of corneal endothelial cells as well as their clinical status. RESULTS: In 23 cases, the number of men and young people under 30 years of age was low, with 4 and 2 cases respectively. There was no visual impairment due to cornea guttata. In the specular microscopic findings, the size and density of dark areas varied according to each case and there was asymmetry in some cases. The parameters of the endothelial cells, comprising mean cell area, hexagonality, and the coefficient of variation, were almost within normal limits except for 1 eye of a 79-year-old man who was speculated to be at an early stage of Fuchs' endothelial corneal dystrophy. Three cases had received cataract surgery, but showed no statistical changes in endothelial morphology before or after the surgery. CONCLUSION: Cornea guttata is thought to be a disease which is not uncommon in Japanese people and which does not bring about visual impairment or abnormalities in corneal endothelial cell parameters except for a small number of patients. Such cases with remarkably decreased cell densities should be considered to be at an early stage of Fuchs' dystrophy. PMID- 10793548 TI - [Histopathological features of the trabecular meshwork in a case of presumed bilateral Chandler syndrome]. AB - BACKGROUND: We describe a patient who had notable decrease in the number of corneal endothelial cells in both eyes and developed open angle glaucoma without evident iris atrophy and peripheral anterior synechia. CASE: A 74-year-old woman. The trabeculectomized angle tissue and iris tissue of her left eye were observed under light and transmission electron microscopy. From Schwalbe's line to the anterior chamber angle, degenerated endothelium-like cells were observed and overgrowth of layers of collagen fiber with varied cycles and basement membrane like material were noticed. In addition, the morphology of intra-trabecular space was atrophic and occlusive with marked degeneration and exfoliation of endothelial cells in the trabecular space. On the side of the anterior chamber, degenerated endothelium-like cells were observed, the morphology of which was considered to result from abnormal metabolic function in corneal endothelium. Overgrowth of basement membrane-like material onto iridic tissue was not observed. CONCLUSION: Although slit lamp examination revealed no obvious abnormality, the result of histological examination suggested that this was a case of Chandler's syndrome in a broad sense or was in the early stage of this disease. We also discussed differential diagnosis from other diseases. PMID- 10793549 TI - [Vaccination for elderly persons]. PMID- 10793550 TI - [Present state and future in organ transplantation]. PMID- 10793551 TI - [Prevention of stroke in Japanese]. PMID- 10793552 TI - [Topics in studies on dementia]. PMID- 10793554 TI - [Oral health care for elderly person]. PMID- 10793553 TI - [Aging, oxygen and the gene]. AB - There are several aspects and approaches to aging research, depending on researchers and their situations. We attempted to elucidate the origin of aging phenomenon but not to investigate the apparent aging phenomenon itself. An important thing to be considered at first was to recognize that the most generally adopted approach to aging research, namely comparison between young and aged men or animals, will give nothing about relationship between the cause and the result of aging. So that we started aging research on genetic basis using the nematode, C. elegans, as experimental animals. This paper gives the story of our investigations over the past 20 years: the planning of investigations that we made before starting actual activity, choice of experimental animal (why the nematode?), the summary of the present state of our investigation and future prospects. In brief, after many difficulties we isolated a mutant of C. elegans, which was named as mev-1. The mutant was found to have a shorter life span than the wild type. In addition, it was hypersensitive to the oxygen atmosphere. The mutant gene was found to correspond to succinate-reductase cytochrome b. In view of the widely accepted evidence, showing intervention of oxygen stress (present research) and perhaps of apoptosis in aging. We can safely say that aging, including natural death, is certainly the result of "life" itself. This contradictory situation might be explainable as, God did not give an eternal advantage to life. PMID- 10793555 TI - [Social subjects in the 21st century and role of the aged]. PMID- 10793556 TI - [Clinical implications of ultrasonic tissue characterization for atherosclerotic carotid intima-media]. AB - Current imaging modalities, such as ultrasonic tissue characterization, have enabled accurate determination of the composition of atherosclerotic plaque of excised aortic tissue. The aim of this study is to compare integrated backscatter (IBS) echo signals of the intima-media complex of human carotid arteries between young and elderly subjects, and to evaluate the ability of ultrasonic tissue characterization. We compared the difference in ultrasonic parameters between the carotid arteries of young healthy subjects (n = 27, 25 +/- 1 y.o.) and elderly subjects (n = 55, 75 +/- 4 y.o.). Intima-media thickness (IMT) and calibrated IBS value (C-IBS: Tissue IBS-vessel lumen IBS value) were measured. The IMT values of young and elderly subjects were 0.54 +/- 0.03 and 1.08 +/- 0.07 mm, respectively (p < 0.01). C-IBS of elderly subjects (11.7 +/- 0.8 dB) was significantly higher than that of young subjects (6.9 +/- 0.7 dB). Significantly wider standard deviation of C-IBS value in each individual was observed in elderly subjects (mean +/- SE of individual standard deviation in C-IBS: 4.2 +/- 0.9 dB) compared with that in young subjects (1.7 +/- 0.3 dB). This might reflect varied composition of atherosclerotic plaques in elderly subjects. IBS analysis would become a promising method to distinguish plaque composition quantitatively and to assess the stability of plaques in clinical setting. PMID- 10793557 TI - [Complications of percutaneous endoscopic gastrostomy in the elderly: local skin infection and respiratory infection]. AB - We investigated post-operative management of acute complications of percutaneous endoscopic gastrostomy (PEG) which often caused respiratory infections and local skin infections. The subjects were a total of 341 patients (male 131, female 210, and the mean age was 80.3), they were classified into six groups by method of feeding and use of antibiotics. Patients were divided into three groups based on the time that feeding was started. In Group I, enteral feeding was not started within the first five days. In Group II, sterilized enteral feeding (lactated Ringer's solution for intravenous infusion) using sterilized intravenous infusion kit started within 24 hours after the procedure, and in Group III, feeding of the usual enteral formula started within 24 hours after the procedure. And as for the using of antibiotics, they were also divided into two groups, antibiotics administered[AB (+)] and no antibiotics administered[AB (-)]. Thus, the patients were divided into six groups according to the time of starting nutrition and the use of antibiotics. The rates of incidence of acute respiratory infections and local skin infections in the six groups were compared by the chi-square test and differences in the rates of incidence of complication were also compared between two PEG methods; the Pull/Push method and the Introducer method. The frequency of local skin infection in Group III was significantly higher than in Group I and Group II. As for the PEG methods, the frequency of local skin infection in the Pull/Push method was significantly higher than Introducer methods. Acute respiratory infections occurred significantly less in the AB (+) group than in the AB (-) group. Postoperative administration of antibiotics would seem to be appropriate for prophylaxis of respiratory infection in elderly patients after PEG. On the other hand, local skin infections are not related to administration of antibiotics, and are highly related to the method of feeding. We concluded that nutrition of sterilized enteral feeding immediately after operation using a sterilized intravenous infusion kit and administration of antibiotics are advisable to prevent major complications in elderly patients. PMID- 10793558 TI - [Feeling of well-being and family contacts in community elderly residents]. AB - The aim of this study is to explore the factors associated with feelings of well being in 683 community-dwelling elderly residents, aged 65 years and over using a self-administered questionnaire. The feeling of well-being was rated using a single question item constructed by The Japanese Association for Cerebro cardiovascular Disease control (JACD), and then subjects were divided into two groups according to answer categories: happiness and unhappiness. There was no statistically significant difference in the proportion of subjects with happiness between males and females, or among age groups. Even when interaction among multiple factors were adjusted by logistic regression analysis, a high frequency of conversation with family was statistically significantly related to happiness, and the odds ratio of it was highest among all other factors. The odds ratio of high frequency of conversation with family was higher among the elderly aged 75 and over than among those aged 65-74. These results suggest that the construction of communication between the elderly and their family through conversation could contribute to feelings of well-being in the elderly. PMID- 10793559 TI - [Epirubicin-based combination chemotherapy combined with G-CSF for the elderly patients with non-Hodgkin's lymphoma]. AB - We devised a new epirubicin-based combination chemotherapy (Epi-COP) regimen for the patients with elderly non-Hodgkin's lymphoma and have treated 30 patients aged 66 years and older who had measurable diseases. In Epi-COP therapy, epirubicin was used as a substitute for doxorubicin in the CHOP regimen, and some dose modifications were made for the other agents. Combined modality treatment (CMT; chemotherapy plus radiotherapy) was adopted for 9 patients with localized disease. Complete response was obtained in 21 of all the 30 patients (70%), 8 in 9 (89%) of the CMT group and 13 in 21 (62%) of the patients with chemotherapy only (chemotherapy group). The median follow up time is 350 days, ranging from 2 to 77 months. The 2 year survival rate was 56% in all patients, 67% in the CMT group and 52% in the chemotherapy group. Granulocyte colony-stimulating factor (G CSF) was administered when the leucocyte count decreased below 2,000/microliter, and 16 patients received it in the first course. The regimen could be repeated every three weeks in most cases. Although we encountered two early deaths, the overall toxicity level seemed to be acceptable. Even when we take account of the small number of patients and the short observation period, it might be concluded that Epi-COP was effective in inducing a good remission rate with moderate toxic effect in elderly patients with non-Hodgkin's lymphoma and CMT should be adopted if it is localized. A randomized comparative study with the CHOP regimen is necessary. PMID- 10793560 TI - [Dementia and disability after initial cerebral thrombosis evaluated by MRI and their clinical course]. AB - This study was conducted to clarify the relationship between long-term prognosis of functional status after stroke and the magnetic resonance imaging (MRI) findings as well as complications in the course. A total of 98 patients with initial cerebral thrombosis were enrolled, and 65 patients surviving 5 years after the event, were studied in terms of cognitive function and activity of daily living (ADL). Mean age at registration (3 months after the event) was 72.0 years-old and 44 were male. MRI findings were divided into eight categories including the size and laterality of infarction, in addition to six categories as previously described. The presence of dementia was identified according to the HDS-R and the DSM-III-R scales, and the extent of dementia, assessed with the CDR scale, was divided into 3 grades: none, mild, and severe. The extent of ADL status was also graded into 3 classes: independent, partially dependent, and completely dependent. Recurrence of stroke, pneumonia, and motor disorders (hip joint fracture) were counted as complications during the course. At baseline, dementia was identified in 44, consisting of 30 mild and 14 severe dementias. During 5 years, 11 cases with mild dementia showed deterioration, and 1 case without dementia developed mild dementia. At registration, there were 46 cases with partially dependent ADL status and 22 cases completely dependent, while deterioration of ADL was seen in 17 during 5 years. Multiple regression analyses showed that diffuse PVH and pneumonia were contributory factors to the development of dementia, while dementia, pneumonia and motor disorders were risk factors for deterioration of ADL. These findings suggested that in patients with cerebral thrombosis, especially in patients with diffuse PVH, pneumonia and motor disorders should be taken care of in order to prevent functional decline. PMID- 10793561 TI - [Portable body cleaning system in bed-bound elderly]. PMID- 10793562 TI - [Molecular bacteriology of Helicobacter pylori]. PMID- 10793563 TI - [Evaluation of chemotherapy for gastric cancer by endoscopic ultrasonography]. AB - We judged the efficacy of chemotherapy using endoscopic ultrasonography (EUS) in 26 cases of gastric cancer. Treatment efficacy was evaluated according to the General Rules for the Gastric Cancer Study, based on the reduction of the largest cross-sectional area of the tumor. A reduction of over 50% was rated as U-PR and a reduction of -25-50% was rated as U-NC. Our findings showed U-PR in 11 cases and U-NC in 15 cases. Three cases initially considered to be NC according to the rules were judged to be U-PR based on EUS findings. Marked therapeutic efficacy in these 3 cases was demonstrated clinically and this was confirmed by EUS findings. Generalized Wilcoxon test showed a significant difference in the cumulative survival rate between U-PR and U-NC cases (p < 0.05). EUS provides an objective means of evaluating the efficacy of chemotherapy in gastric cancer patients, including those with lesions that cannot be evaluated by the General Rules for the Gastric Cancer Study. PMID- 10793564 TI - [A patient with gastric cancer accompanied by high levels of serum AFP, CEA, CA19 9 and CA125]. PMID- 10793565 TI - [A case of gastric mucosal carcinoma with remarkable infiltration of lymphoid cells negative for Epstein-Barr virus]. PMID- 10793566 TI - [A case of ulcerative colitis with skipped lesion in right-sided colon and rectum]. PMID- 10793568 TI - [A case of Brunner's gland hamartoma which is consistent of fat tissue and Brunner's gland hyperplasia]. PMID- 10793567 TI - [A case of ulcerative colitis complicated with venous thrombosis and pulmonary embolism]. PMID- 10793569 TI - [A case of early syphilitic hepatitis with repetitive right hypochondriac pain]. PMID- 10793570 TI - [A case of hepatic adenomatous hyperplasia which was enhanced with computed tomography during arterial portography (CTAP)]. PMID- 10793571 TI - [A case of successful TIPS placement for gastrointestinal hemorrhage from portal hypertensive gastropathy due to complete portal venous thrombosis]. PMID- 10793572 TI - [A Japanese case of idiopathic hemochromatosis with analysis of HFE gene mutations and a review of literature on HLA phenotypes in the Japanese cases]. PMID- 10793573 TI - [A case of gallbladder cancer producing granulocyte-colony stimulating factor and possible parathyroid hormone related protein]. PMID- 10793574 TI - [Pulsatile flow detection for evaluation of tumor vessels of hepatocellular carcinoma]. PMID- 10793575 TI - [Identification of two distinct types of four-transmembrane domain proteins, occludin and claudins: towards new physiology in paracellular pathway]. PMID- 10793576 TI - [Chemistry of oxygen reduction by cytochrome c oxidase--resonance Raman spectroscopic study]. PMID- 10793577 TI - [D-amino acids in nature]. PMID- 10793578 TI - [Neuronal survival and glial cells]. PMID- 10793579 TI - [Hematopoietic development--from cytokine research to discovery of hemangioblast]. PMID- 10793580 TI - [Translesion DNA polymerases]. PMID- 10793581 TI - [Physiological function and mechanism of action of neuropeptide PACAP]. PMID- 10793582 TI - [Mechanism of ligand independent activation of Ah receptor]. PMID- 10793583 TI - [Transglutaminase and apoptosis]. PMID- 10793584 TI - [Simple cultivation method for anaerobic hyperthermophiles]. PMID- 10793585 TI - Postoperative residual curarization and evidence-based anaesthesia. PMID- 10793586 TI - Neuropsychometric deficit after cardiac surgery: a new approach for a new millennium. PMID- 10793587 TI - Placement of double lumen tubes--time to shed light on an old problem. PMID- 10793588 TI - Effect of chronic intercurrent medication with beta-adrenoceptor blockade or calcium channel entry blockade on postoperative silent myocardial ischaemia. AB - We have examined observational data from four published studies investigating the incidence of postoperative silent myocardial ischaemia (post-SMI) for the effects of chronic intercurrent therapy with beta-adrenoceptor blockade or chronic calcium channel entry blockade. A total of 453 patients underwent ambulatory ECG monitoring before and for 2 days after non-cardiac surgery; 79 patients were receiving chronic intercurrent beta-adrenoceptor blockade and 70 calcium channel entry blockade for ischaemic heart disease or arterial hypertension. Using logistic regression analysis, we defined a model for post-SMI that included four significant terms: beta-adrenoceptor blockade; calcium channel entry blockade; arterial hypertension; and vascular surgery. Using univariate regression, there was no effect of chronic beta-adrenoceptor blocking therapy on post-SMI (odds ratio 0.94 (95% confidence intervals 0.54-1.65)), but there was a higher incidence of post-SMI in patients receiving chronic calcium channel entry blocking drugs (odds ratio 1.95 (1.15-3.32); P = 0.015). There was no interaction between beta-adrenoceptor blockade and calcium channel entry blockade for postoperative SMI (odds ratio 2.48 (0.71-8.73)), but there was an interaction between beta-adrenoceptor blockade, calcium channel entry blockade, hypertension and vascular surgery (P = 0.0201). These findings are at variance with those which have shown effects of preoperative beta-adrenoceptor blockade on the incidence of post-SMI over the first 7 days after operation, and on mortality rates to 2 yr. There are no comparable data examining the effects of chronic intercurrent calcium channel entry blockade. PMID- 10793589 TI - Effect of midazolam on transfer function between beat-to-beat arterial pressure and inter-beat interval length. AB - Arterial pressure (AP) and inter-beat interval (IBI) length are under autonomic nervous system control. The control mechanisms can be investigated by transfer function analysis. It is not known if this type of analysis may be helpful in monitoring depth of sedation. In an open-label, uncontrolled investigation, the effect of midazolam on the transfer function between AP and IBI, and on spectral indices of AP and heart rate (HR) variability (APV, HRV) were assessed in the absence and presence of the benzodiazepine antagonist flumazenil. We studied 11 healthy male volunteers. After an initial control period of 60 min, we studied three consecutive periods, each of 60 min duration, with progressively increasing concentrations of midazolam (0.02, 0.06, 0.14 mg kg-1 h-1). A final 60-min period during administration of flumazenil 0.004 mg kg-1 h-1 and while the agonist was still present was also studied. To confirm midazolam-induced central nervous system effects, electroencephalography was performed and Ramsay sedation scores were determined. With increasing dose of midazolam, the high frequency (0.15-0.4 Hz) component of the transfer function between AP and IBI decreased progressively (mean 26.5 (SEM 3.7), 19.2 (2.9), 12.8 (1.7), 8.4 (1.6) ms mm Hg-1). This effect was antagonized by flumazenil (21.5 (3.2) ms mm Hg-1). Other indices (e.g. HRV, APV) did not reveal such a clear response to midazolam dose and flumazenil application. Thus in healthy male volunteers, the transfer function between AP and IBI in the parasympathetically dominated high frequency range varies according to benzodiazepine agonism and antagonism. This finding has potential implications for monitoring the effects of benzodiazepines. PMID- 10793590 TI - Biphasic changes in autonomic nervous activity during pregnancy. AB - To understand the sequential response of the autonomic nervous system to pregnancy, we studied heart rate variability in 23 first trimester, 23 second trimester and 21 third trimester pregnant women. Twenty non-pregnant women were recruited as controls. Time and frequency domain measures of heart rate variability in three recumbent positions were compared. We found that normalized high-frequency power in the supine position increased significantly in the first trimester (42.2 (95% confidence interval (CI) 5.4) nu (normalized unit); P < 0.05) compared with non-pregnant controls (33.0 (6.0) nu), and then decreased progressively in the second (27.3 (6.7) nu) and third (21.8 (6.0) nu; P < 0.05) trimesters. The low-/high-frequency power ratio in the supine position decreased significantly in the first trimester (0.8 (0.3); P < 0.05) compared with that of non-pregnant controls (1.1 (0.3)) and increased progressively in the second (1.5 (0.4)) and third (2.1 (0.8); P < 0.05) trimesters. When the position was changed from the supine to the right lateral decubitus, the percentage change in normalized high-frequency power correlated significantly and negatively with normalized high-frequency power in the supine position in non-pregnant controls (r = -0.56, P = 0.01) and in pregnant women in the first (r = -0.44, P = 0.034), second (r = -0.68, P < 0.001) and third (r = -0.68, P < 0.001) trimesters. These results indicate that autonomic nervous activity shifted towards a lower sympathetic and higher vagal modulation in the first trimester, and changed towards a higher sympathetic and lower vagal modulation in the third trimester as gestational age increased. The balance between the haemodynamic changes of pregnancy and aortocaval compression caused by the enlarging gravid uterus may be responsible for the biphasic changes in autonomic nervous activity during pregnancy. PMID- 10793591 TI - Transoesophageal echocardiographic assessment of haemodynamic changes during laparoscopic herniorrhaphy in small children. AB - Laparoscopic techniques for surgery are gradually becoming established in paediatric surgery. Technical aspects, such as the maximum safe gas insufflation pressure, are still open to discussion. We used transoesophageal echocardiography to study the haemodynamic changes in eight small children undergoing laparoscopic herniorrhaphy, with two different levels of intra-abdominal pressure (IAP), 6 and 12 mm Hg. End-tidal carbon dioxide tension was maintained constant at 4.3-4.7 kPa. After baseline measurements, an IAP of 12 mm Hg was applied for 10 min. Next, IAP was decreased to 6 mm Hg, followed by a second period of 12 mm Hg. Haemodynamic measurements were obtained at each stage. A further measurement was obtained 10 min after abdominal deflation at the end of surgery while anaesthesia was unchanged. Cardiac index (CI) decreased significantly only after the first 12 mm Hg level of IAP. The subsequent decrease in IAP to 6 mm Hg caused return of CI to baseline levels. The second increase in IAP did not cause any reduction in CI. The initial reduction in CI, although statistically significant, did not appear to be clinically important. We conclude that an IAP of up to 12 mm Hg appeared to be safe in healthy small children undergoing laparoscopic herniorrhaphy. PMID- 10793592 TI - Oral preanaesthetic medication for children: double-blind randomized study of a combination of midazolam and ketamine vs midazolam or ketamine alone. AB - Anxiolysis and sedation with oral midazolam are common practice in paediatric anaesthesia. However, good or excellent results are seen in only 50-80% of cases. For this reason, we investigated if addition of a low dose of oral ketamine (MIKE: ketamine 3 mg kg-1, midazolam 0.5 mg kg-1) resulted in better premedication compared with oral midazolam 0.5 mg kg-1 or ketamine 6 mg kg-1 alone, in a prospective, randomized, double-blind study. We studied 120 children (mean age 5.7 (range 2-10) yr) undergoing surgery of more than 30 min duration. After oral premedication in the ward and transfer, the child's condition in the induction room was evaluated by assigning 1-4 points to the quality of anxiolysis, sedation, behaviour at separation from parent and during venepuncture (transfer score). On days 1 and 7 after operation, parents were interviewed for changes in behaviour (eating, sleep, dreams, toilet training), recollection and satisfaction, using a standardized questionnaire. The groups were similar in age, sex, weight, intervention and duration of anaesthesia. The transfer score was significantly better in the MIKE group (12.5 (95% confidence interval (CI) 11.9 13.1)) than in the ketamine or midazolam groups (10.6 (9.8-11.4) and 11.5 (10.7 12.3), respectively). Success rates for anxiolysis and behaviour at separation were greater than 90% with the combination, approximately 70% with midazolam and only 51% with ketamine alone. The incidence of salivation, excitation and psychotic symptoms was low in all groups. Vertigo and emesis before induction were significantly more frequent after ketamine premedication. During recovery, there were no differences in sedation or time of possible discharge. After 1 week, parents reported nightmares (ketamine five, midazolam three, MIKE one), restless sleep (five/four/four) or negative memories (three/four/one). There were no major or continuing disturbances in behaviour or development. In summary, significantly better anxiolysis and separation were observed with a combination of ketamine and midazolam, even in awake children (sedation was not successful according to the preset criteria), than with midazolam or ketamine alone. Duration of action and side effects of the combination were similar to those of midazolam. The combination of both drugs in strawberry flavoured glucose syrup (pH 4.5 approximately) is chemically stable for 8 weeks. PMID- 10793593 TI - S(+)-ketamine for caudal block in paediatric anaesthesia. AB - We have evaluated the intra- and postoperative analgesic efficacy of preservative free S(+)-ketamine compared with bupivacaine for caudal block in paediatric hernia repair. After induction of general anaesthesia, 49 children undergoing hernia repair were given a caudal injection (0.75 ml kg-1) of S(+)-ketamine 0.5 mg kg-1 (group K1), S(+)-ketamine 1.0 mg kg-1 (group K2) or 0.25% bupivacaine with epinephrine 1:200,000 (group B). No additional analgesic drugs were required during operation in any of the groups. Haemodynamic and respiratory variables remained stable during the observation period. Mean duration of analgesia was significantly longer in groups B and K2 compared with group K1 (300 (SD 96) min and 273 (123) min vs 203 (117) min; P < 0.05). Groups B and K2 required less analgesics in the postoperative period compared with group K1 (30% and 33% vs 72%; P < 0.05). Postoperative sedation scores were comparable between the three groups. We conclude that S(+)-ketamine 1.0 mg kg-1 for caudal block in children produced surgical and postoperative analgesia equivalent to that of bupivacaine. PMID- 10793594 TI - Pharmacokinetics of tramadol in children after i.v. or caudal epidural administration. AB - We have studied the pharmacokinetics of a single bolus dose of tramadol 2 mg kg-1 injected either i.v. or into the caudal epidural space in 14 healthy children, aged 1-12 yr, undergoing elective limb, urogenital or thoracic surgery. Serum concentrations of tramadol and its metabolite O-demethyl tramadol (MI) were measured in venous blood samples at various intervals up to 20 h by non stereoselective gas chromatography with nitrogen-selective detection. All pharmacokinetic variables were evaluated using a non-compartmental model. After a single i.v. injection (n = 9), the mean elimination half-life of tramadol was 6.4 (SD 2.7) h, with a volume of distribution of 3.1 (1.1) litre kg-1 and total plasma clearance of 6.1 (2.5) ml kg-1 min-1. All of these pharmacokinetic variables were similar to those reported previously in adults. After caudal epidural administration (n = 5), mean elimination half-life was 3.7 (0.9) h, volume of distribution was 2.0 (0.4) litre kg-1 and total clearance was 6.6 (1.9) ml kg-1 min-1. The caudal/i.v. quotient of the AUC was 0.83, which confirms that there is extensive systemic absorption of tramadol after caudal administration. Serum concentrations of MI showed a time course typical of a metabolite after both modes of administration. Serum concentrations of MI after caudal administration were lower than those after i.v. injection. PMID- 10793595 TI - Comparison of compound A concentrations with sevoflurane anaesthesia using a closed system with a PhysioFlex anaesthesia machine vs a low-flow system with a conventional anaesthesia machine. AB - Sevoflurane anaesthesia was conducted using a totally closed circuit PhysioFlex anaesthesia machine (PhysioFlex group) or with a standard Modulus CD anaesthesia machine (Modulus group) (n = 8 in each group). The PhysioFlex was used under closed system conditions and the Modulus was used under low-flow system conditions (flow rate 1 litre min-1). Concentrations of sevoflurane degradation products and the temperature of soda lime were compared. Degradation products in the circuit were measured hourly, and the temperature of soda lime was monitored. The only degradation product detected was CF2 = C(CF3)-O-CH2F (compound A). Maximum concentrations of compound A were significantly lower (median 8.5 (range 5.4-15.9) ppm) in the PhysioFlex than in the Modulus group (21.2 (16.5-27.4) ppm) (P < 0.05). The maximum temperature of soda lime was also significantly lower in the PhysioFlex group (35.3 (32.1-36.3) degrees C vs 44.6 (43.0-47.1) degrees C, respectively) (P < 0.05). Hourly compound A concentrations were lower in the PhysioFlex group than in the Modulus group. End-tidal sevoflurane concentrations during measurement of degradation products were not different between groups. Therefore, use of the totally closed PhysioFlex system may significantly reduce compound A concentrations compared with low-flow anaesthesia using a standard anaesthesia machine. PMID- 10793596 TI - Effects of the sitting position on the distribution of blood volume in patients undergoing neurosurgical procedures. AB - The use of the sitting position in neurosurgery is often associated with decreased arterial pressure (MAP) and stroke volume index (SVI). A shift in blood from the intra- to the extrathoracic compartment may be responsible for this cardiovascular response. However, little is known of the amount of shift in blood volume after transfer from the supine to the sitting position. Therefore, we measured simultaneously changes in intrathoracic blood volume (ITBV) caused by a change in body position in anaesthetized patients. Measurements of cardiac index (CI), ITBV, pulmonary (PBV) and total circulating (TBVcirc) blood volumes were performed in the supine and sitting position. CI, ITBV, PBV and TBVcirc were measured using a thermodye dilution technique. Fluid input was restricted to 14 ml kg-1 before induction of anaesthesia. Change in body position caused a significant decrease in ITBV and was accompanied by a significant decrease in CI, SVI and MAP. Changes in ITBV correlated (r = 0.78) with changes in SVI. Thus a change in blood volume distribution between the intra- and extrathoracic compartment occurred after a change from the supine to the sitting position. Indicator dilution enables quantification of this shift and may be helpful in guiding fluid therapy in selected patients. PMID- 10793598 TI - Effect of cricoid pressure on insertion of and ventilation through the cuffed oropharyngeal airway. AB - We have assessed the effect of cricoid pressure on insertion of and ventilation through the cuffed oropharyngeal airway (COPA) in 53 patients, in a double-blind, randomized study. Two anaesthetists assessed adequacy of ventilation in anaesthetized and paralysed patients at the same time but using different methods. The first assessed ventilation clinically, by observing synchronized chest expansion with gentle manual ventilation and the second noted measurements of tidal volume (VT) and peak inspiratory pressure (PIP). Five mask ventilated breaths ('baseline') were assessed as above. Patients were then allocated randomly to receive cricoid pressure (group A, n = 28) or no cricoid pressure (group B, n = 25). Five further mask ventilated breaths ('after manoeuvre') were again assessed. A COPA was then inserted and five further breaths ('after COPA') were assessed. A COPA was inserted at the first attempt in all patients except for one in group A who required two attempts. COPA placement was difficult in one patient in group B who had a small distance between the incisor teeth. Ventilation was clinically 'adequate' in all patients except for one in the cricoid pressure group. There were no significant differences in measured VT or PIP between 'baseline' and 'after manoeuvre' breaths. Significant differences in VT and PIP were found after COPA insertion in the group that received cricoid pressure, with a mean decrease in VT of 108 ml (P = 0.0049) and a mean increase in PIP of 5.2 cm H2O (P = 0.0111). PMID- 10793597 TI - Effects of fenoterol and ipratropium on respiratory resistance of asthmatics after tracheal intubation. AB - We have studied the effects of a beta-agonist, fenoterol, and a cholinergic antagonist, ipratropium, on post-intubation total respiratory system resistance (Rrs) in asthmatics who developed increased Rrs after tracheal intubation. Sixteen stable asthmatics in whom Rrs increased after intubation were allocated randomly to receive either 10 puffs of fenoterol (group F) or 10 puffs of ipratropium (group IB) via a metered dose inhaler 5 min after intubation. Anaesthesia was induced and maintained with propofol i.v. Rrs was recorded before treatment and again 5, 15 and 30 min after treatment. Rrs decreased significantly from pretreatment values by mean 53 (SD 8)%, 53 (7)% and 58 (6)% at 5, 15 and 30 min, respectively, in group F, but declined by only 12 (6)%, 15 (4)% and 17 (5)% in group IB. At all times after treatment, patients in the fenoterol group had significantly lower Rrs values than those in the ipratropium group. We conclude that increased Rrs after tracheal intubation in asthmatics can be reduced effectively by treatment with fenoterol. A secondary finding of our study was that even after induction of anaesthesia with propofol, patients with a history of asthma may develop high Rrs. PMID- 10793599 TI - Efficacy of ginger for nausea and vomiting: a systematic review of randomized clinical trials. AB - Ginger (Zingiber officinale) is often advocated as beneficial for nausea and vomiting. Whether the herb is truly efficacious for this condition is, however, still a matter of debate. We have performed a systematic review of the evidence from randomized controlled trials for or against the efficacy of ginger for nausea and vomiting. Six studies met all inclusion criteria and were reviewed. Three on postoperative nausea and vomiting were identified and two of these suggested that ginger was superior to placebo and equally effective as metoclopramide. The pooled absolute risk reduction for the incidence of postoperative nausea, however, indicated a non-significant difference between the ginger and placebo groups for ginger 1 g taken before operation (absolute risk reduction 0.052 (95% confidence interval -0.082 to 0.186)). One study was found for each of the following conditions: seasickness, morning sickness and chemotherapy-induced nausea. These studies collectively favoured ginger over placebo. PMID- 10793600 TI - Effects of halothane on respiratory mechanics and lung histopathology in normal rats. AB - It is generally accepted that halothane reduces airway and tissue resistance in lungs with preexisting airway tone. However, under conditions of resting airway tone, pulmonary resistance remains unaltered. In this study, we have determined the effects of halothane on respiratory system, pulmonary and chest wall resistive, elastic and viscoelastic mechanical properties, and related the results to findings from lung histology in intact normal rats. Sixteen adult male Wistar rats were allocated randomly to one of two groups (n = 8 in each group): control or halothane group. In the control group, animals were sedated with diazepam 5 mg i.p. and anaesthetized with pentobarbital 20 mg kg-1 i.p. In the halothane group, the anaesthetic was administered at an end-tidal concentration of I MAC throughout the study. Rats were paralysed and underwent mechanical ventilation. Halothane decreased airway resistance but increased the tissue component of resistance (caused by viscoelastic elements and lung inhomogeneity). Static and dynamic elastance also increased with halothane anaesthesia. Pulmonary resistance remained unchanged. Lung histopathology demonstrated airway dilatation and a greater degree of lung collapse and hyperinflation in the halothane group. We conclude that halothane anaesthesia acts both on airway and lung tissue. In airway tissue, dilatation occurs but the lung periphery stiffens. Consequently, these opposing effects result in no overall apparent change in mechanical properties, although changes are observed during halothane anaesthesia in normal animal and subjects. PMID- 10793601 TI - Central nervous system complications of cardiac surgery. AB - The neurological complications of cardiac surgery are associated with significantly increased mortality, morbidity and resource utilization. The use of new surgical techniques, introduction of wider indications for surgery and increased public expectation has led to an increase in the average age of cardiac surgical patients and an increased incidence of repeat procedures. With these changes has come an increased risk of neurological complications. The likelihood of perioperative stroke varies between 1% and 5% in most published series and is dependent on a multitude of risk factors. Of these, patient age, aortic atheroma, symptomatic cerebrovascular disease, diabetes mellitus and the type of surgery appear to be most important. Cognitive deterioration after cardiac surgery is far more common, affecting as many as 80% of patients a few days after surgery and persisting in one-third. Despite an increase in the age of the cardiac surgical population, the reported incidence of cognitive dysfunction after cardiac surgery seems to have fallen in recent years. Whether this is a real phenomenon or the result of changes in the use of psychometric testing and the definition of cognitive decline remains unclear. Recognition that certain equipment, surgical practices and patient factors contribute to neurological morbidity has prompted 'neuroprotective' interventions. Some of these (e.g. arterial line filtration and alpha-stat management) have been shown to improve outcome. Despite these measures, a small number of patients will inevitably sustain cerebral injury during otherwise successful cardiac surgery. Although pharmacological neuroprotection may, in the future, offer some of these patients an improved outcome, it is unlikely that any single agent will prevent neurological injury. In the meantime, the CNS complications of cardiac surgery remain a fertile area of research. PMID- 10793602 TI - Residual curarization in the recovery room after vecuronium. AB - We have investigated residual block after anaesthesia which included the use of the neuromuscular blocking agent vecuronium but no anticholinesterase, in 568 consecutive patients on admission to the recovery room. The ulnar nerve was stimulated submaximally using TOF stimulation (30 mA). Postoperative residual curarization was defined as a TOF ratio < 0.7. Of the 568 patients, 239 (42%) had a TOF < 0.7 in the recovery room. These patients had received a larger cumulative dose of vecuronium than patients who had full recovery (mean 7.7 (SD 3.6) mg vs 6.2 (2.7) mg; P < 0.05) and a shorter time had elapsed since the last vecuronium dose (117 (70) min vs 131 (80) min; P < 0.05). Of 435 patients whose trachea was extubated, 145 (33%) exhibited inadequate recovery from neuromuscular block. Six of these had one or no response to TOF stimulation and were reintubated. In the remaining 139 patients, neuromuscular block was successfully antagonized. Only 20 patients (3.5%) remembered TOF stimulation when questioned 2 h later in the recovery room, and discomfort associated with it was assessed using a visual analogue scale before discharge. We conclude that it is necessary to antagonize residual block produced by vecuronium. PMID- 10793604 TI - Warm air sensation for assessment of block after spinal anaesthesia. AB - We have evaluated a new method of assessing dermatomal sensory levels after regional anaesthesia based on warm sensation. Sensory levels were assessed in 30 patients after spinal anaesthesia using a respiratory gas humidifier, adapted to deliver a constant flow of warm air at 40 +/- 0.2 degrees C. This was compared with the cold sensation from ethyl chloride spray. The frequency distribution of the dermatomal differences showed 96.6% of the comparisons were between +1 and -1 dermatomes. The median difference in dermatomal levels between the two methods of assessment was 0 (interquartile range 0-1) (P = 0.65). We conclude that the warm air method compares favourably with ethyl chloride spray and both can be used interchangeably. PMID- 10793603 TI - Effect of intraoperative angiotensin-converting enzyme inhibition by quinaprilat on hypertension after coronary artery surgery. AB - Activation of the renin-angiotensin system during cardiopulmonary bypass (CPB) may be involved in early postoperative hypertension after coronary artery bypass grafting (CABG). As hypertensive episodes may be deleterious in the immediate postoperative period, we have assessed the effects of prophylactic treatment with the angiotensin-converting enzyme inhibitor quinaprilat in an open study. During steady state CPB, patients received quinaprilat 0.02 mg kg-1 (group A, n = 10), quinaprilat 0.04 mg kg-1 (group B, n = 10) or saline solution (group C, n = 10) as an i.v. bolus dose. Sodium nitroprusside (SNP) was given after operation when systolic arterial pressure was > 150 mm Hg. Requirements for SNP 1 h after arrival in the ICU were significantly less in groups A (two of 10) and B (two of 10) than in group C (eight of 10). Also, patients in group C had a greater systolic arterial pressure compared with groups A and B. There were no significant differences between groups in diastolic arterial pressure, heart rate, cardiac index or cardiac filling pressures. We conclude that quinaprilat can be used during CABG to reduce the incidence of postoperative hypertension. Further studies of the efficacy and safety of this technique are necessary. PMID- 10793605 TI - Caudal clonidine for postoperative analgesia in adults. AB - We have assessed the analgesic efficacy and side effects of caudally administered clonidine in a prospective, randomized, double-blind, placebo-controlled study. We studied 64 adult patients undergoing elective haemorrhoidectomy. Caudal block was performed in all patients using a mixture of 0.5% bupivacaine 35 mg with 2% lidocaine 140 mg and epinephrine 5 micrograms ml-1. Patients were allocated randomly to one of two groups. Clonidine 75 micrograms was added in group C and saline 1 ml in group S. Median time to first analgesic requirements was significantly longer in group C (mean 729 (SD 120) min) than in group S (276 (131) min) (P = 0.01). Bradycardia occurred in seven patients in group C but did not affect mean arterial pressure. PMID- 10793606 TI - Preparation for regional anaesthesia induces changes in thrombelastography. AB - The effects of crystalloid and colloid infusions on coagulation measured by thrombelastography (TEG) present a confused picture. The influence of environmental factors may explain the disparity between previous studies. We studied two groups of 20 women presenting at term for elective Caesarean section. In the first group, TEG analysis was performed before and after infusion of Gelofusine 500 ml over 15 min. The second group was treated in the same way except that subjects did not receive fluid. We found significant changes in r and k values in both groups, suggesting enhanced coagulation. As hypercoagulable changes were also seen in the group that did not receive fluid preload, the hypothesis that moderate haemodilution causes hypercoagulability must be questioned. The influence of environmental factors can explain differences reported between in vivo and in vitro studies. PMID- 10793607 TI - Rectally administered dimenhydrinate reduces postoperative vomiting in children after strabismus surgery. AB - We have investigated the effectiveness of rectally administered dimenhydrinate on postoperative vomiting in children undergoing strabismus surgery, in a double blind, randomized, placebo-controlled study. In one group, dimenhydrinate 50 mg was administered rectally 30 min before starting anaesthesia, whereas in the control group, placebo suppositories were given. Children who received dimenhydrinate showed a significantly (P < 0.001) lower incidence of vomiting (15%) than those in the control group (75%). We conclude that rectal administration of dimenhydrinate is an effective means of reducing postoperative vomiting in children undergoing strabismus surgery. PMID- 10793608 TI - Postpartum headache after epidural blood patch: investigation and diagnosis. AB - Use of an epidural blood patch to treat spinal headache after accidental dural puncture is well recognized. The high success rate associated with this practice has been questioned and it is not uncommon for patients to suffer recurring headaches after a supposedly successful blood patch. We describe a patient in labour who suffered accidental dural puncture, and whose headache was treated twice with an epidural blood patch. Despite this, the headache persisted. The case highlights the difficulty in the diagnosis of headache in the postnatal period in patients who have had regional analgesia and the importance of considering an alternative pathology, even if epidural blood patching has been successful. In this case, a diagnosis of cortical vein thrombosis was made. The incidence, presentation, aetiology and treatment of this rare condition is described. PMID- 10793609 TI - Patient-controlled analgesia in labour using remifentanil in two parturients with platelet abnormalities. AB - Two term parturients with documented platelet abnormalities presented to the delivery suite in labour. Because regional analgesic techniques were contraindicated, we elected to use patient-controlled i.v. remifentanil for pain relief. The patient-controlled analgesia (PCA) device was programmed to give a bolus dose of remifentanil 20 micrograms over 20 s with a lockout time of 3 min, and no background infusion. Analgesia was reported as very good by the mothers and by the attending midwives. There were no adverse neonatal sequelae. If there are facilities to monitor the neonate and mother, this method of analgesia may prove useful in those patients where regional techniques are not possible, but further research is needed to ascertain its safety and appropriateness in such circumstances. PMID- 10793610 TI - Safe use of remifentanil in a patient treated with the monoamine oxidase inhibitor phenelzine. AB - We report the safe use of remifentanil as part of the anaesthetic technique in a patient undergoing major head and neck surgery who was being treated for depressive illness with the non-specific monoamine oxidase inhibitor (MAOI) phenelzine. PMID- 10793611 TI - Exhaustion of an ABEK nitric oxide absorber. PMID- 10793612 TI - Costs of sevoflurane and propofol anaesthesia. PMID- 10793614 TI - Adverse effects of cannabis and cannabinoids. PMID- 10793613 TI - Costs of sevoflurane and propofol anaesthesia. PMID- 10793616 TI - Acoustic monitoring for neuromuscular block. PMID- 10793615 TI - Pulmonary aspiration of gastric contents in obstetrics. PMID- 10793617 TI - Bradycardia after administration of remifentanil. PMID- 10793618 TI - Pulmonary haemorrhage after percutaneous paravertebral block. PMID- 10793619 TI - Complications of paravertebral block. PMID- 10793620 TI - Induction of superovulation by inhibin vaccine in cyclic guinea-pigs. AB - Experiments were conducted to determine whether neutralizing endogenous inhibin affects follicular development and ovulation rate in guinea-pigs. Eighteen female guinea-pigs bearing 4 week progesterone implants were divided into three groups. At 1 week after removal of the progesterone implants, the animals were given a s.c. injection of 1 ml placebo (saline in oil emulsion; control), or 25 or 50 micrograms inhibin vaccine three times at 4 week intervals. Blood samples were collected once a week throughout the experiment for measuring inhibin antibody titres. After the third injection of inhibin vaccine, blood samples and ovaries were collected on the morning of day 8 after the day of oestrus. Inhibin vaccine increased the ovulation rate in a dose-dependent manner (placebo: 4.2 +/- 0.4; 25 micrograms inhibin vaccine: 6.2 +/- 0.9; 50 micrograms inhibin vaccine: 9.8 +/- 0.9) without any effects on the duration of the oestrous cycle. The results also showed that active immunization against inhibin increased the number of atretic follicles of 300-399 microns in diameter on day 8 after ovulation. The present study is the first to show that the active immunization against inhibin may be a useful method for inducing multiple ovulation in guinea-pigs. PMID- 10793621 TI - Effect of exogenous LH pulses on the fate of the first dominant follicle in postpartum beef cows nursing calves. AB - Prolonged postpartum anoestrus in beef cows is due to failure of early dominant follicles to ovulate. It is hypothesized that this failure to ovulate is due to inadequate LH pulse frequency. The objective of this study was to determine whether administration of hourly LH pulses would cause the first dominant follicle to ovulate. In Expt 1, 16 cows received either saline (n = 8) or porcine LH (pLH; 50 micrograms h-1; n = 8) as hourly pulses for 3-5 days from the second day of dominance of the first dominant follicle (day 0). In Expt 2, 21 cows received either saline (n = 7), or 50 micrograms pLH (n = 7) or 100 micrograms pLH (n = 7) as hourly pulses for 3 days. Appropriate ovarian scanning and assays of blood samples were carried out. In Expt 1, the number of dominant follicles that underwent atresia was not affected by increasing the number of LH pulses, but the duration of dominance (days) of the first and second dominant follicles and maximum size (mm) of the second dominant follicle were increased (P < 0.05). Oestradiol concentrations were higher (P < 0.05) in cows given hourly pLH pulses (3.1 +/- 1.2 pg ml-1) compared with controls (1.2 +/- 0.2 pg ml-1). Four of eight treated cows had an anovulatory LH surge. The number of follicle waves to first ovulation was not different (P < 0.05) between control (4.6 +/- 0.9) and pLH treated cows (3.9 +/- 0.5). In Expt 2, four of seven cows given pulses of 100 micrograms pLH h-1 ovulated the first dominant follicle, and the interval from calving to first ovulation was decreased (P < 0.05). In the remaining three cows, the duration of dominance of the first dominant follicle was increased (P < 0.005), the maximum size of the first dominant follicle was greater (P < 0.05), and the interval (days) from the start of infusion to new wave emergence was greater (P < 0.05) compared with cows that failed to ovulate in either the 50 micrograms pLH h-1 or control treatments. In conclusion, hourly pulses of pLH from day 1 after dominance of the first dominant follicle in postpartum beef cows can either prolong dominance or induce it to ovulate. This finding supports the hypothesis that LH pulse frequency is a key determinant of the fate of the dominant follicle in the early postpartum period. PMID- 10793623 TI - Effects of an organophosphate pesticide, quinalphos, on the hypothalamo-pituitary gonadal axis in adult male rats. AB - The effects of chronic sub-lethal doses (7-14 mg kg-1 a day for 15 days) of quinalphos were evaluated in adult male rats for changes in testicular morphology, circulatory concentrations of hormones (LH, FSH, prolactin and testosterone), activities of acetylcholinesterase (AChE) and angiotensin converting enzyme (ACE) as well as metabolism of biogenic amines (dopamine, noradrenaline and 5-hydroxytryptamine (5-HT)) in the hypothalamus and pituitary. Hormones were assayed by radioimmunoassay or chemiluminescent immunoassay (testosterone). The enzymes were estimated after spectrophotometry and the biogenic amines by HPLC-electrochemistry. Sub-lethal chronic administration of quinalphos resulted in: decreased testicular mass and AChE activity in central as well as peripheral organs; increased serum LH, FSH, prolactin and testosterone concentrations; decreased pituitary or increased testicular ACE activity; severe disruption of spermatogenesis with increasing doses of pesticide; and no significant effects on dopamine, noradrenaline or 5-HT concentrations in the hypothalamus or pituitary. Administration of oestradiol (50 micrograms per rat a day) during pesticide treatment resulted in: a significant decrease in the mass of the testis and accessory sex organs; decreases in serum LH, FSH, testosterone concentrations; an increase in prolactin concentration; and a decrease in dopamine or an increase in noradrenaline and 5-HT in the hypothalamus or pituitary. Oestradiol had a marked effect: in pesticide-treated animals, the pesticide effects were significantly reversed. This indicates that in pesticide toxicity, the hypothalamo-pituitary-gonadal axis is operational. Since many of the observed pesticide effects could be inhibited by oestradiol, it is suggested that the pesticide acts directly on the gonadotrophins. In conclusion, quinalphos decreases fertility in adult male rats by affecting the pituitary gonadotrophins. PMID- 10793622 TI - Involvement of matrix metalloproteinases 2 and 9, tissue inhibitor of metalloproteinases and apoptosis in tissue remodelling in the sheep placenta. AB - Placental growth and development is crucial for successful pregnancy. The aim of this study was to characterize the activity and localization of the matrix metalloproteinase 2 (MMP-2) and MMP-9, which are capable of degrading basement membrane collagen (predominantly collagen type IV), and their endogenous tissue inhibitor of matrix metalloproteinases (TIMPs), in amniotic fluid and in the developing ovine placenta. Cell deletion by apoptosis during placental development was also examined. Zymography with gelatin as substrate indicated that MMP-2 (72 kDa gelatinase A; predominantly latent form) was present in increasing amounts in amniotic fluid from day 70 of gestation to labour (days 140 145), and MMP-9 (92 kDa gelatinase B; predominantly latent form) was detectable from day 125 to labour; there was no increase in MMP-2 or -9 in labour. A broad range of TIMPs was detected in amniotic fluid; the molecular masses corresponded to TIMP-1, -2 and -3. Immunohistochemical techniques localized MMP-2, MMP-9 and TIMP-3 in the sheep placenta, predominantly in the trophoblast layer in uninucleate, but not binucleate, cells. However, MMP-2 and -9 activated proteins in placental homogenates were low throughout pregnancy. Apoptosis was identified by morphological criteria and also by TdT-mediated dUTP nick end labelling. Apoptosis was present in discrete regions in the placenta, predominantly in trophoblast cells near the tips and the basal regions of the fetomaternal interdigitations. During pregnancy the sheep placenta becomes more complex and the area of the fetomaternal interface increases. MMP-2 and -9 are likely to be involved in breaking down basement membranes to allow cell migration during this process. It is suggested that digestion of supporting extracellular matrix may trigger apoptosis and in some way increase the branching pattern in the villi. PMID- 10793624 TI - Effect of acute immunoneutralization of endogenous leptin on prolactin and LH secretion during the afternoon of pro-oestrus or in steroid-treated ovariectomized female rats. AB - Recent data indicate that leptin is involved in the control of reproductive function. Experiments were carried out to analyse the role of endogenous leptin in the regulation of LH and prolactin secretion during the afternoon of pro oestrus and that induced by ovarian steroids in ovariectomized rats. In the first experiment, cyclic female rats were implanted with intra-auricular and intracerebroventricular (i.c.v.) cannulae and, at pro-oestrus, were injected (i.c.v.) with 10 microliters normal rabbit serum or leptin antiserum (at 13:00 and 14:00 h). Blood samples were obtained at 10:00 h and at intervals of 1 h between 13:00 and 20:00 h. In the second experiment, female rats in pro-oestrus were injected with normal rabbit serum or leptin antiserum at 16:00 and 18:00 h and blood samples were taken every 10 min between 18:00 and 20:00 h. In the third experiment, adult female rats that had been ovariectomized 2 weeks before were implanted with intra-auricular and i.c.v. cannulae and treated with oestradiol benzoate (30 micrograms s.c.) at 10:00 h and progesterone (2 mg s.c.) 48 h later. Normal rabbit serum (10 microliters) or leptin antiserum (10 microliters) were injected (i.c.v.) at 13:00 and 14:00 h, and blood samples were obtained at 10:00 h and at intervals of 1 h between 13:00 and 20:00 h. In the fourth experiment, hemipituitaries from ovariectomized steroid-treated female rats were incubated in the presence of leptin116-130 (an active fragment of the native molecule), GnRH or leptin + GnRH. Prolactin and LH secretion during the afternoon of pro-oestrus in females treated with leptin antiserum was similar to that observed in animals injected with normal rabbit serum. In ovariectomized female rats, the steroid induced LH surge increased slightly after administration of leptin antiserum, whereas the prolactin surge remained unchanged. In vitro, leptin116-130 (10(-5) to 10(-8) mol l-1) inhibited LH secretion and modulated the effect of GnRH on LH release, depending on the concentration of GnRH: leptin116-130 (10(-6) mol l-1) reduced the effectiveness of 10(-7) mol GnRH l-1 and increased that of 10(-9) mol GnRH l-1. In conclusion, these experiments indicate that acute immunoneutralization of endogenous leptin does not interfere with spontaneous or steroid-induced LH and prolactin surges. In addition, the finding that leptin116 130 inhibited LH release and modulated the effectiveness of GnRH in vitro provides evidence of the direct modulatory role of leptin on LH secretion acting at the pituitary. PMID- 10793625 TI - Effect of inhibitors and uncouplers of oxidative phosphorylation during compaction and blastulation of bovine embryos cultured in vitro. AB - The effect of inhibiting ATP production via oxidative phosphorylation during pericompaction of in vitro produced bovine embryos was investigated. This was achieved by: (i) varying the atmospheric O2 concentration (0, 1, 2, 4 and 7%); (ii) addition of oxidative phosphorylation inhibitors, NaN3 and antimycin A; and (iii) addition of 2,4-dinitrophenol, an uncoupler of oxidative phosphorylation from electron transport. The development of embryos under various O2 concentrations from day 5 to day 7 of development indicated that an optimal concentration occurred at about 2%. Addition of NaN3 revealed that doses above 100 mumol l-1 were toxic to embryo development, but that concentrations of 5-10 mumol l-1 stimulated embryo development by 10-25%. A similar result was observed after addition of 2,4-dinitrophenol, whereas antimycin A was inhibitory at doses as low as 1 mumol l-1. At concentrations of NaN3 or 2,4-dinitrophenol that stimulated embryo development, the number of cells of the resulting blastocysts was also significantly increased. Addition of NaN3 from day 1 of development inhibited subsequent development. Metabolic data of NaN3-treated embryos revealed that O2 uptake was significantly lower at inhibitory doses (100 mumol l-1). A significant (P < 0.05) log linear increase in glucose uptake was measured between the three concentrations of NaN3 (0, 10 and 100 mumol l-1). These results demonstrate that ATP production via oxidative phosphorylation is essential for bovine embryo development in vitro. However, transient (subacute) inhibition appears to be beneficial to embryo development and the number of cells, perhaps by creating a more favourable intracellular environment. PMID- 10793626 TI - Stimulation of Ca(2+)-dependent exocytosis of the sperm acrosome by cAMP acting downstream of phospholipase A2. AB - Spermatozoa undergo exocytosis in response to agonists that induce Ca2+ influx and, in turn, activation of phosphoinositidase C, phospholipase C, phospholipase A2, and cAMP formation. Since the role of cAMP downstream of Ca2+ influx is unknown, this study investigated whether cAMP modulates phospholipase C or phospholipase A2 using a ram sperm model stimulated with A23187 and Ca2+. Exposure to dibutyryl-cAMP, phosphodiesterase inhibitors or forskolin resulted in enhancement of exocytosis. However, the effect was not due to stimulation of phospholipase C or phospholipase A2: in spermatozoa prelabelled with [3H]palmitic acid or [14C]arachidonic acid, these reagents did not enhance [3H]diacylglycerol formation or [14C]arachidonic acid release. Spermatozoa were treated with the phospholipase A2 inhibitor aristolochic acid, and dibutyryl-cAMP to test whether cAMP acts downstream of phospholipase A2. Under these conditions, exocytosis did not occur in response to A23187 and Ca2+. However, inclusion of dibutyryl-cAMP and the phospholipase A2 metabolite lysophosphatidylcholine did result in exocytosis (at an extent similar to that seen when cells were treated with A23187/Ca2+ and without the inhibitor). Inclusion of lysophosphatidylcholine alone, without dibutyryl-cAMP, enhanced exocytosis to a lesser extent, demonstrating that cAMP requires a phospholipase A2 metabolite to stimulate the final stages of exocytosis. These results indicate that cAMP may act downstream of phospholipase A2, exerting a regulatory role in the exocytosis triggered by physiological agonists. PMID- 10793627 TI - Effects of superovulated heifer diet type and quantity on relative mRNA abundances and pyruvate metabolism in recovered embryos. AB - This study investigated the effects of quantity and type of diet fed to superovulated donor heifers on molecular and metabolic indices of embryonic development. These effects included the relative abundances of mRNAs for the alpha 1 subunit of Na/K-ATPase and the antioxidant enzyme Cu/Zn-SOD, as well as pyruvate utilization in bovine morulae and blastocysts developed in vivo. Heifers were fed a daily ration of either grass silage and a citrus-beet pulp-based concentrate or grass silage and a barley-based concentrate for 116 days, both at 3 kg per day or ad libitum. In embryos derived from heifers fed the pulp-based diets, the relative abundances of the transcripts were not affected by either day of collection or quantity of diet. In embryos derived from heifers fed the barley based diets, the relative abundances of the Na/K-ATPase transcripts were also not changed by these main effects, while the relative abundances of the Cu/Zn-SOD transcripts were affected by day of collection and by the quantity of diet. Pyruvate metabolism was affected by day of collection, and was significantly increased in day 8 embryos compared with day 7 and day 6 embryos. Diet quantity did not affect pyruvate utilization, whereas diet type did increase pyruvate metabolism in the barley group when compared with the pulp group. The results of this study show for the first time that molecular and metabolic variations may exist in embryos derived in vivo and developed in donor heifers on nutritional regimens differing in type and quantity. Differences in embryos collected on different developmental days may be attributed to varying cell numbers. Alterations in the relative abundances of the Cu/Zn-SOD transcripts and pyruvate metabolism caused by the quantity of diet fed to the donor animal were likely to have been due to alterations in metabolic end products that accumulate in reproductive tract fluids, whereas differences in embryonic metabolism caused by type of diet are related to the composition of the diet. These findings characterize embryos produced in vivo at the molecular level, indicating that the molecular markers used in the present study can differentiate between populations of embryos produced under different nutritional regimens and determine conditions conductive to the production of good quality embryos. PMID- 10793628 TI - Evaluation of cholesteryl ester transfer in the seminiferous tubule cells of immature rats in vivo and in vitro. AB - Sertoli cells and germ cells are separated from the interstitial blood capillaries by an extracellular matrix and the peritubular cells, which constitute a barrier to the movement of plasma lipoproteins. The present study was undertaken to evaluate in vivo and in vitro the high density lipoprotein (HDL) cholesteryl ester transfer from plasma to seminiferous tubule cells in the testis of 30-day-old rats. Firstly, the transfer of HDL cholesteryl oleate from plasma to testicular compartments was evaluated and, secondly, the role of apolipoproteins A-I and E in the uptake of cholesteryl ester by Sertoli cells was investigated. At 2 h after the administration of HDL reconstituted with [3H]cholesteryl ester, dimyristoyl phosphatidylcholine and apolipoproteins, the tissue space in the interstitial cells (740 +/- 60 microliters g-1 cell protein) was fourfold higher than that in the seminiferous tubule cells (170 +/- 10 microliters g-1). Sertoli cells were isolated and incubated with [3H]cholesteryl ester HDL reconstituted with apolipoprotein A-I or E to evaluate the mechanisms of cholesteryl ester influx. At the same apolipoprotein concentration (50 micrograms apolipoprotein ml-1 medium), the uptake of [3H]cholesteryl oleate from phospholipid-apolipoprotein E vesicles was twofold higher than that with phospholipid-apolipoprotein A-I vesicles. The presence of heparin reduced the uptake of cholesteryl ester from apolipoprotein E vesicles but not with apolipoprotein A-I vesicles, indicating that uptake of apolipoprotein A-I vesicles via a secretion of apolipoprotein E by the cells themselves was not involved. These results demonstrate that plasma lipoprotein cholesterol is able to cross the testis lamina propria and that Sertoli cells take up cholesteryl ester for seminiferous tubule cell metabolism mainly via an apolipoprotein E pathway. PMID- 10793629 TI - Effects of the Booroola Fec gene on ovarian follicular populations in superovulated Romanov ewes pretreated with a GnRH antagonist. AB - Endocrine control of follicular growth was studied in mature Romanov ewes carrying (RF+) or not carrying (R+2) the Booroola Fec gene during an oestrous cycle after gonadotrophin-dependent follicles were suppressed by treatment with an antagonist of GnRH (Antarelix, 0.5 mg per day) and superovulatory treatment was administered. The left ovary was removed after 10 days of treatment (saline or Antarelix) and the right ovary was removed at the end of the superovulatory treatment. Ewes of both genotypes treated with Antarelix had lower plasma LH concentrations than did controls from day 0 to day 10. The inhibitory effect of Antarelix on LH concentration increased with day of treatment. The variability in FSH concentrations during the initial 10 days was reduced by Antarelix treatment in both genotypes. Plasma FSH concentrations were higher in RF+ ewes than in R+2 ewes. In both genotypes, FSH concentrations varied significantly with day of treatment, with the lowest concentrations at day 8 and the highest concentrations at day 5. RF+ ewes had a greater total and atretic number of antral follicles 0.62-1.12, 1.12-2.00 and 2.00-3.00 mm in diameter (classes 2, 3 and 4) than did R+2 ewes before and after superovulatory treatment. After superovulatory treatment, the total number of atretic and non-atretic follicles > 3.00 mm in diameter (class 5) increased in both genotypes. Superovulatory treatment also increased the number of total and atretic class 4 follicles in RF+ only. Conversely, superovulatory treatment decreased the mean number of class 3 follicles in both genotypes, while the number of atretic follicles was decreased only in R+2 ewes. Antarelix treatment significantly reduced the percentage of follicles > 2.00 mm in diameter in RF+ but not in R+2 ewes. Antarelix treatment before superovulatory treatment increased the total number of class 4 follicles in both genotypes but the increase was more significant in RF+ than in R+2 ewes. These results indicate that Antarelix pretreatment favours a greater superovulatory response in Romanov ewes carrying the Fec gene because ovulatory follicles are recruited from a wider range of follicular size classes. PMID- 10793630 TI - Hysteroscopic insemination of small numbers of spermatozoa at the uterotubal junction of preovulatory mares. AB - Mares were inseminated with motile spermatozoa suspended in 30-150 microliters Tyrode's medium directly onto the uterotubal papilla at the anterior tip of the uterine horn, ipsilateral to the ovary containing a dominant preovulatory follicle of > or = 35 mm in diameter, by means of a fine gamete intrafallopian transfer (GIFT) catheter passed through the working channel of a strobed light videoendoscope. Insemination of 10, 8, 25, 14, 11 and 10 mares with, respectively, 10.0, 5.0, 1.0, 0.5, 0.1 or 0.001 x 10(6) motile spermatozoa resulted in conception rates of, respectively, 60, 75, 64, 29, 22 and 10%. Deposition of 1.0 x 10(6) motile spermatozoa onto the uterotubal papilla began to approach the limit of successful fertilization. These doses are far lower than the 3-15 x 10(9) spermatozoa normally ejaculated by fertile stallions during mating, and the accepted minimum dose of 500 x 10(6) spermatozoa used for conventional uterine body insemination in mares. The simplicity of the technique offers a practical means of exploiting new breeding technologies that require very small numbers of spermatozoa in horse breeding. PMID- 10793631 TI - Reproductive hormone profiles in mares during the autumn transition as determined by collection of jugular blood at 6 h intervals throughout ovulatory and anovulatory cycles. AB - The aim was to define precisely the FSH secretion pattern in mares during the two ovulatory cycles before, and for 24 days after, the last ovulation of the season and to compare this with the profiles of other reproductive hormones and follicular growth to identify changes which may lead to the termination of follicular cycles. Jugular blood was collected every 6 h from ten light horse mares for 6 weeks in autumn. Samples were assayed for FSH, LH, prolactin, inhibin, oestrone conjugates and progesterone. Luteolysis occurred earlier and periovulatory oestrone, but not inhibin, concentrations were significantly lower in the last than in the second to last cycles. In ovulatory and anovulatory cycles, daily mean FSH concentrations were low at the expected time of ovulation and high between days 9 and 11 (day 0 = ovulation), which were usually after luteolysis. However, the periovulatory FSH nadir was prolonged in the last compared with the second to last cycles, and the difference between peak and trough values was not significant in anovulatory cycles. Between day 5 and day 8, the FSH interpulse interval was approximately 2 days, and did not vary in successive cycles. The LH profile also showed progressive changes as mares entered acyclicity; the surge terminated sooner in the last than in the second to last cycles, and failed to occur when expected in acyclicity. Sporadic prolactin pulses occurred at luteolysis in a similar proportion of ovulatory and anovulatory cycles. These results indicate that inadequate gonadotrophin stimulation in early dioestrus may be a critical event leading to suboptimal follicular and luteal development, and eventually acyclicity. Moreover, the time relationships amongst changes in pituitary and ovarian hormones and follicular growth become increasingly disrupted during the autumn transition, which may contribute to the cessation of cyclicity. PMID- 10793632 TI - Effect of nitric oxide synthase inhibitors on ovum transport and oviductal smooth muscle activity in the rat oviduct. AB - The effect of the inhibition of nitric oxide synthase (NOS) on ovum transport and oviductal motility in rats was investigated. Three different NOS inhibitors were injected into the ovarian bursa at oestrus or day 3 of pregnancy. Oviducts and uteri were flushed 24 h later and the presence of ova was recorded. In oestrous and pregnant rats, treatment resulted in accelerated egg transport, as shown by a decrease in the number of ova present in the oviducts. In cyclic rats, intrabursal injection of 1 mg kg-1 of either N-monomethyl-L-arginine (L-NMMA) or N omega nitro-L-arginine methyl ester (L-NAME) elicited a 30% reduction in the number of ova present in the oviducts, whereas in pregnant animals, the same dose of L-NMMA produced a reduction of 40%. Simultaneous administration of the NO donor spermine NONOate (5 mg kg-1) completely reversed the effect of L-NMMA. Tubal motility was assessed by microsphere displacement analysis within the oviduct. Surrogate ova were transferred to the oviductal lumen at oestrus and 24 h later the effect of intraoviductal injection of 1 microgram L-NMMA or vehicle was assessed. The microspheres in the isthmus showed an oscillating motion, and periods in which movement was not detectable. However, L-NMMA treatment produced a 3.6-fold increase in the maximum instant velocities and a significant reduction in the resting periods of the microspheres compared with the control group (P < 0.001). These results provide evidence that NO inhibition increases tubal motility that results in accelerated ovum transport, and indicate that NO could act as a paracrine signal between different layers of the oviductal wall, providing a role for endogenous NO in regulation of tubal function. PMID- 10793633 TI - Effects of activin A and follistatin on developmental kinetics of bovine embryos: cinematographic analysis in a chemically defined medium. AB - The effects of recombinant human activin A and follistatin on the developmental kinetics of bovine presumptive zygotes matured and fertilized in vitro using time lapse cinematography were investigated. The presumptive zygotes were cultured for 9 days in a chemically defined medium (modified synthetic oviduct fluid, control) and modified synthetic oviduct fluid supplemented with activin A or follistatin. Development under cine-recording conditions was similar to that in an incubator. Addition of activin A to modified synthetic oviduct fluid increased, while addition of follistatin decreased, the percentage of zygotes that developed to morulae and blastocysts. Follistatin significantly prolonged the timing of development to the 9-16-cell stage compared with the control and activin A media. Activin A significantly shortened the duration of the third cell cycle compared with the control, but follistatin significantly prolonged the fourth cell cycle compared with the control and activin A. Developmental arrest ('lag-phase') during the 4-8-cell stage was observed in 95% of embryos developed to more than the 9-16-cell stage in all treatments. The greater the number of cells at the onset of the lag-phase, the earlier the onset of the phase and the shorter the duration of the phase, the further embryos were able to develop by day 9 in all treatments. The number of cells at the onset of the lag-phase in the medium containing activin A was significantly higher than it was in control or follistatin-containing media. Moreover, activin A significantly shortened the duration of the lag-phase compared with follistatin. The present results indicate that activin A may enhance in vitro development of bovine embryos by improving developmental kinetics, especially by increasing the number of cells at the onset of the lag-phase and shortening the duration of this phase. PMID- 10793634 TI - Effect of ornidazole on fertility of male rats: inhibition of a glycolysis related motility pattern and zona binding required for fertilization in vitro. AB - The effects of the male antifertility agent ornidazole on glycolysis as a prerequisite for fertilization were investigated in rats. Antifertility doses of ornidazole inhibited glycolysis within mature spermatozoa as determined from the lack of glucose utilization, reduced acidosis under anaerobic conditions and reduced glycolytic enzyme activity. As a consequence, cauda epididymidal spermatozoa from ornidazole-fed rats were unable to fertilize rat oocytes in vitro, with or without cumulus cells, which was not due to transfer of an inhibitor in epididymal fluid with the spermatozoa. Under IVF conditions, binding to the zona pellucida was reduced in spermatozoa from ornidazole-fed males and the spermatozoa did not undergo a change in swimming pattern, which was observed in controls. The block to fertilization could be explained by the disruption of glycolysis-dependent events, since reduced binding to the zona pellucida and a lack of kinematic changes were demonstrated by control spermatozoa in glucose free media in the presence of respiratory substrates. The importance of glycolysis for binding to, and penetration of, the zona pellucida, and hyperactivation in rats is discussed in relation to the glycolytic production of ATP in the principal piece in which local deprivation of energy may explain the reduced force of spermatozoa from ornidazole-fed males. PMID- 10793635 TI - Effect of GnRH antagonist-induced prolonged follicular phase on follicular atresia and oocyte developmental competence in vitro in superovulated heifers. AB - A GnRH antagonist (Antarelix) was used to suppress endogenous pulsatile secretion of LH and delay the preovulatory LH surge in superovulated heifers to study the effect of a prolonged follicular phase on both follicle and oocyte quality. Oestrous cycles were synchronized in 12 heifers with progestagen (norgestomet) implants for 10 days. On day 4 (day 0 = day of oestrus), heifers were stimulated with 24 mg pFSH for 4 days and luteolysis was induced at day 6 with PGF2 alpha (2 ml Estrumate). Animals in the control group (n = 4) were killed 24 h after the last FSH injection. At this time, heifers in group A36h (n = 4) and group A60h (n = 4) were treated with 1.6 mg of Antarelix every 12 h for 36 and 60 h, respectively, and then killed. After dissection of ovarian follicles, oocytes were collected for individual in vitro maturation, fertilization and culture; follicular fluid was collected for determination of steroid concentrations, and granulosa cells were smeared, fixed and stained for evaluation of pycnosis rates. Granulosa cell smears showed that 90% of follicles were healthy in the control group. In contrast, 36 and 58% of the follicles in group A36h showed signs of early or advanced atresia, respectively, while 90% of the follicles in group A60h showed signs of late atresia. Intrafollicular concentrations of oestradiol decreased (P < 0.0001) from healthy follicles (799.14 +/- 40.65 ng ml-1) to late atretic follicles (3.96 +/- 0.59 ng ml-1). Progesterone concentrations were higher (P < 0.0001) in healthy follicles compared with atretic follicles, irrespective of degree of atresia. Oestradiol:progesterone ratios decreased (P < 0.0001) from healthy (4.58 +/- 0.25) to late atretic follicles (0.07 +/- 0.009). The intrafollicular concentrations of oestradiol and progesterone were significantly higher (P < 0.0001) in the control than in the treated groups. The oestradiol:progesterone ratio was higher (P < 0.0001) in the control (4.55 +/- 0.25) than in the A36h (0.40 +/- 0.05) and A60h (0.07 +/- 0.009) groups. Unexpectedly, the cleavage rate of fertilized oocytes, blastocyst rate and number of cells per blastocyst were not significantly different among control (85%, 41% and 95 +/- 8), A36h (86%, 56% and 93 +/- 5) and A60h (88%, 58% and 79 +/- 4) groups. In addition, there were no significant differences in the blastocyst rates from oocytes derived from healthy (45%), early atretic (54%), advanced atretic (57%) and late atretic follicles (53%). In conclusion, the maintenance of the preovulatory follicles in superovulated heifers with a GnRH antagonist induced more atresia and a decrease in oestradiol and progesterone concentrations. However, the developmental potential in vitro to day 8 of the oocytes recovered from these atretic follicles was not affected. PMID- 10793636 TI - Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability. AB - This study was designed to assess whether nucleic acid stains such as propidium iodide and Hoechst 33258 and the cytosolic stain eosin identified equivalent proportions of non-viable cells. Sub-samples of boar spermatozoa stored for up to 72 h, and frozen bull spermatozoa stored in straws and thawed before staining, were exposed to either propidium iodide or Hoechst 33258 alone or in combination. Additional sub-samples were stained with eosin-nigrosin and subsequently with Giemsa. The proportion of non-viable cells identified by propidium iodide alone was equivalent to that observed when it was used in combination with the other fluorescent probe. Similar results were observed for Hoechst 33258. However, direct microscopic examination of sub-samples exposed to both stains revealed that a proportion of spermatozoa stained with propidium iodide did not incorporate Hoechst 33258. This was found consistently in boar and bull spermatozoa under the different experimental conditions used. Quantification showed that the proportion of propidium iodide-positive cells was significantly higher than Hoechst 33258-positive cells. Furthermore, the proportion of propidium iodide-positive cells was higher than cells stained with eosin, but no differences were found between the number of cells stained with Hoechst 33258 or eosin. The proportion of cells stained with propidium iodide was positively correlated with the proportion stained with either Hoechst 33258 or eosin, despite the observation that more cells incorporated propidium iodide. Taken together, these results indicate that there are differences in the ability of fluorescent probes to identify non-viable sperm cells and that this should be considered when staining protocols are used to analyse sperm viability, or when viability is used as a discriminating factor in functional studies, such as those related to acrosomal exocytosis. PMID- 10793637 TI - Relationship between different stages of the corpus luteum and the expression of the peroxisome proliferator-activated receptor gamma protein in bovine large lutein cells. AB - Lutein cells produce progestins that support pregnancy. Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism. Peroxisome proliferator-activated receptors (PPAR) are transcription factors that are central in the regulation of lipid metabolism. Hence, they may play a role in regulation of the development and regression of the corpus luteum. The present study investigated the expression of PPAR-gamma, n during different stages of the corpus luteum. Lutein cells were isolated mechanically from non-pregnant and pregnant heifers on days 5, 12 and 20 of the oestrous cycle (n = 3 for each day). PPAR-gamma in single cells was analysed by flow cytometry. PPAR-gamma 1 and PPAR gamma 2 isoforms were distinguished by immunoblotting. The cell cycle of the lutein cells was measured by the flow cytometric quantification of DNA in single cells, using propidium iodide staining after ethanol fixation and RNAse treatment, and by the detection of the proliferating cell nuclear antigen (PCNA). The response of the cells to PPAR-gamma agonist 15-deoxy-delta 12,14 prostaglandin J2 (15dPGJ2, 200 and 490 nmol l-1) with and without changing the cell cycle by the anti-apoptogenic drug aurintricarboxylic acid (ATA, 10 mumol l 1) was used as an in vitro model to study the relationship between the cell cycle and PPAR-gamma. The concentration of PPAR-gamma per cell from non-pregnant heifers was significantly higher on day 5 (3.40 +/- 0.30 fmol) compared with that on day 12 (1.34 +/- 0.18 fmol, P < 0.05) and day 20 (0.55 +/- 0.2 fmol, P < 0.05). In pregnant heifers, the concentration of PPAR-gamma was significantly (P < 0.01) higher than in non-pregnant heifers. A decrease in the PPAR-gamma 1 isoform relative to PPAR-gamma 2 was observed in cells on day 12 of the oestrous cycle compared with day 5. The cell cycle (S phase portion in cells on days 5, 12 and 20: 16 +/- 4%, 6 +/- 4% and 4 +/- 3%, respectively) and the portion of cells with PCNA correlated with the amount of PPAR-gamma in non-pregnant heifers. ATA promoted the S phase in cells of non-pregnant heifers (day 12) and the endogenous agonist of PPAR-gamma, 15dPGJ2, inhibited the response to ATA in a dose-dependent manner, indicating that PPAR-gamma plays a role in the arrest of the cell cycle in lutein cells to maintain their differentiated state. PMID- 10793638 TI - Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. AB - Cattle, pig and sheep oocytes isolated from healthy cumulus-oocyte complexes were pooled, within species, to provide samples of immature denuded oocytes with intact zona pellucida (n = 1000 per sample) for determination of fatty acid mass and composition in total lipid, constituent phospholipid and triglyceride. Acyl containing lipid extracts, transmethylated in the presence of a reference penta decaenoic acid (15:0), yielded fatty acid methyl esters which were analysed by gas chromatograph. Mean (+/- SEM) fatty acid content in samples of pig oocytes (161 +/- 18 micrograms per 1000 oocytes) was greater than that in cattle (63 +/- 6 micrograms; P < 0.01) and sheep oocytes (89 +/- 7 micrograms; P < 0.05). Of 24 fatty acids detected, palmitic (16:0; 25-35%, w/w), stearic (18:0; 14-16%) and oleic (18:1n-9; 22-26%) acids were most prominent in all three species. Saturated fatty acids (mean = 45-55%, w/w) were more abundant than mono- (27-34%) or polyunsaturates (11-21%). Fatty acids of the n-6 series, notably linoleic (18:2n 6; 5-8%, w/w) and arachidonic acid (20:4n-6; 1-3%), were the most abundant polyunsaturates. Phospholipid consistently accounted for a quarter of all fatty acids in the three species, but ruminant oocytes had a lower complement of polyunsaturates (14-19%, w/w) in this fraction than pig oocytes (34%, w/w) which, for example, had a three- to fourfold greater linoleic acid content. An estimated 74 ng of fatty acid was sequestered in the triglyceride fraction of individual pig oocytes compared with 23-25 ng in ruminant oocytes (P < 0.01). It is concluded that the greater fatty acid content of pig oocytes is primarily due to more abundant triglyceride reserves. Furthermore, this species-specific difference, and that in respect of polyunsaturated fatty acid reserves, may underlie the contrasting chilling, culture and cryopreservation sensitivities of embryos derived from pig and ruminant (cattle, sheep) oocytes. PMID- 10793639 TI - Effect of inhibition of sterol delta 14-reductase on accumulation of meiosis activating sterol and meiotic resumption in cumulus-enclosed mouse oocytes in vitro. AB - Two sterols of the cholesterol biosynthetic pathway induce resumption of meiosis in mouse oocytes in vitro. The sterols, termed meiosis-activating sterols (MAS), have been isolated from human follicular fluid (FF-MAS, 4,4-dimethyl-5 alpha cholest-8,14,24-triene-3 beta-ol) and from bull testicular tissue (T-MAS, 4,4 dimethyl-5 alpha-cholest-8,24-diene-3 beta-ol). FF-MAS is the first intermediate in the cholesterol biosynthesis from lanosterol and is converted to T-MAS by sterol delta 14-reductase. An inhibitor of delta 7-reductase and delta 14 reductase, AY9944-A-7, causes cells with a constitutive cholesterol biosynthesis to accumulate FF-MAS and possibly other intermediates between lanosterol and cholesterol. The aim of the present study was to evaluate whether AY9944-A-7 added to cultures of cumulus-oocyte complexes (COC) from mice resulted in accumulation of MAS and meiotic maturation. AY9944-A-7 stimulated dose dependently (5-25 mumol l-1) COC to resume meiosis when cultured for 22 h in alpha minimal essential medium (alpha-MEM) containing 4 mmol hypoxanthine l-1, a natural inhibitor of meiotic maturation. In contrast, naked oocytes were not induced to resume meiosis by AY9944-A-7. When cumulus cells were separated from their oocytes and co-cultured, AY9944-A-7 did not affect resumption of meiosis, indicating that intact oocyte-cumulus cell connections are important for AY9944-A 7 to exert its effect on meiosis. Cultures of COC with 10 mumol AY9944-A-7 l-1 in the presence of [3H]mevalonic acid, a natural precursor for steroid synthesis, resulted in accumulation of labelled FF-MAS, which had an 11-fold greater amount of radioactivity incorporated per COC compared with the control culture without AY9944-A-7. In contrast, incorporation of radioactivity into the cholesterol fraction was reduced 30-fold in extracts from the same oocytes. The present findings demonstrate for the first time that COC can synthesize cholesterol from mevalonate and accumulate FF-MAS in the presence of AY9944-A-7. Furthermore, AY9944-A-7 stimulated meiotic maturation dose dependently, indicating that FF MAS, and possibly other sterol intermediates of the cholesterol synthesis pathway, play a central role in stimulating mouse oocytes to resume meiosis. The results also indicate that oocytes may not synthesize steroids from mevalonate. PMID- 10793640 TI - Effects of inhibitors of arachidonic acid turnover on the production of prostaglandins by the guinea-pig uterus. AB - The supply of free arachidonic acid from phospholipids is generally regarded as the rate-limiting step for prostaglandin (PG) synthesis by tissues. Two enzymes involved in arachidonic acid uptake into, and release from, phospholipids are acyl-CoA:lysophospholipid acyltransferase (ACLAT) and phospholipase A2 (PLA2), respectively. PGF2 alpha produced by the endometrium induces luteolysis in several species including guinea-pigs. Thimerosal, an inhibitor of ACLAT, and aristolochic acid, an inhibitor of PLA2, both reduced, in a concentration dependent manner, the output of PGF2 alpha from guinea-pig endometrium cultured for 24 h on days 7 and 15 of the oestrous cycle. This study showed that the continual production of PGF 2 alpha by guinea-pig endometrium is not only dependent upon the activity of PLA2 for releasing free arachidonic acid for PGF2 alpha synthesis, but also on the incorporation of arachidonic acid into the phospholipid pool by the activity of ACLAT. The inhibitory effects of thimerosal and aristolochic acid on the outputs of PGE2 and 6-keto-PGF1 alpha were less marked, particularly on day 7 when the low output of PGE2 was unaffected and the output of 6-keto-PGF1 alpha was increased at the lower concentrations of thimerosal. This finding indicates that there are different pools of arachidonic acid bound as phospholipid for the syntheses of PGF2 alpha and 6-keto-PGF1 alpha by guinea-pig endometrium. PMID- 10793642 TI - Ontogeny of oestrogen receptor alpha in gonads and sex ducts of fetal and newborn mice. AB - The distribution of nuclear oestrogen receptor alpha (ER-alpha) in the sex organs of fetal and newborn mice has been investigated immunohistochemically. There was no visible ER-alpha immunoreaction in the sexually undifferentiated gonads, whereas a faint immunoreaction was detected in a few cells surrounding the sex ducts, the Mullerian and Wolffian ducts. After sex differentiation, the immunoreaction of ER-alpha was observed in various cell types, with the exception of both the male and female germ cells. In the fetal ovary, immunoreaction was restricted to the surface epithelium and a few stroma cells without any preferential localization. In the testis, the number of ER-alpha-immunopositive cells, identified as Leydig and peritubular cells, increased with age. Immediately after sex differentiation, cells surrounding the sex ducts were ER alpha-immunopositive, but no immunoreaction was detected in the epithelium in either sex. During development, the epithelium of the sex ducts attained a topographic difference in ER-alpha immunoreaction. In females, immunoreaction was detected in the epithelium of the oviduct, but not in the uterus. In males, the immunoreaction of ER-alpha was intense in the epithelium of the efferent ducts, weak in the epididymis and absent in the vas deferens. ER-alpha immunoreaction in the cells surrounding the sex duct differed between the sexes, being high in all these cells in females, but of varying intensity in males. ER-alpha may not play an important role in the development and function of ovarian cells, but may be involved in the development of Leydig and peritubular cells. Furthermore, substances that react with ER-alpha may influence the male germ cells indirectly through the ER-alpha-immunopositive peritubular cells. In addition, in both sexes, ER-alpha-immunopositive cells surrounding the sex ducts may be involved in the mediation of growth and functional differentiation of the ductal epithelium. PMID- 10793641 TI - Heterogeneity in the bioactivity of LH secreted by peripubertal rats. AB - Treatment of immature rats with 5 iu equine chorionic gonadotrophin (eCG) on day 25 typically stimulates a preovulatory surge of LH on day 27 and ovulation on day 28. In rats weighing > 60 g at the time of treatment, an LH surge and ovulation occurred in 75% of the animals but, in rats weighing < 60 g, only 13% ovulated even though 69% showed an LH surge. Previous findings have shown that exogenous LH can stimulate ovulation in the rats < 60 g, indicating that the anovulation was not due to ovarian immaturity, but rather to an abnormal form of LH. Thus, it was important to determine whether the bioactivity of LH released at the time of the surge differs in rats < 60 g compared with rats > 60 g. Experiments showed that LH from both groups of eCG-treated animals were equipotent in stimulating testosterone production from incubated Leydig cells and progesterone production from cultured granulosa cells. Similarly the surge of progesterone in vivo, which occurs co-incident with the LH surge, was of similar magnitude in both groups of animals. Since prostaglandin synthesis increases at the time of ovulation and is also stimulated by LH, it was investigated whether the activity of ovarian phospholipase A2, the rate limiting enzyme in prostaglandin synthesis, and ovarian prostaglandin E2 concentrations differed in the animals > 60 g and < 60 g. Phospholipase A2 activities were similar in both groups of animals at the time of the LH surge, as were the prostaglandin E2 concentrations. However, in all animals that ovulated (15/20 in rats > 60 g and 2/15 in rats < 60 g), there was a threefold increase in ovarian prostaglandin E2 concentrations. The results show that, in underweight animals, the bioactivity of LH, in terms of its ability to stimulate steroidogenesis and phospholipase A2 activity, is similar to that released by animals > 60 g; however, the LH produced by the underweight animals fails to induce ovulation by failing to increase, either directly or indirectly, prostaglandin E2 production. Comparison of the profiles of plasma LH collected at the time of the LH surge on an anionic ion exchange column indicates that the LH from rats < 60 g possesses significantly less of the neutral or basic glycoform of LH than that from rats > 60 g. This finding provides a further index that the biopotency of LH produced by underweight animals is different from that of rats > 60 g. PMID- 10793643 TI - Effect of long-term selection for early postnatal growth rate on survival and prenatal development of transferred mouse embryos. AB - Reciprocal embryo transfer procedures were performed among mouse selection lines to examine prenatal maternal effects on survival and development of transferred embryos. Mice were from generations 28 and 29 of an experiment to select for (i) increased body weight again from 0 to 10 days (E+); (ii) decreased body weight gain from 0 to 10 days (E-); or (iii) a randomly bred control line (C). A total of 118 embryo transfer procedures performed 12 h after conception resulted in 983 progeny born to 89 litters. There was a 39% overall embryo survival rate and 75% overall pregnancy success rate. Response to superovulation and oestrous synchronization was significantly lower (P < 0.01) in the E+ line. E+ individuals that did superovulate produced an average of 37 oocytes per flush, which was significantly higher than in the control line mice (29 oocytes per flush; P < 0.01). The ability to complete pregnancy successfully was not influenced by uterine environment or embryo-uterine interaction. In contrast, embryo survival in successful pregnancies was significantly affected by uterine environment. There were large maternal effects for body weight and tail length at birth; E+ recipients produced pups that were significantly larger than E- recipient pups (P < 0.01), which in turn were significantly larger than pups gestated by control recipients (P < 0.01). PMID- 10793644 TI - Obesity in North America. An overview. AB - The terms "obesity" and "overweight" mean different things to different people. This article discusses such issues as prevalence, morbidity, mortality, and psychosocial effects. Definitions and various classifications of obesity are discussed also. PMID- 10793645 TI - Genetic and environmental contributions to obesity. AB - Groups that are already concerned about obesity can be linked together. This can generate plans of action and may lead to small-scale interventions. At the same time, awareness needs to be raised among other potential partners, especially federal, state, and local government agencies. As a partnership is built, funding opportunities, from public and private entities, should increase. The ultimate need is a public health campaign to combat an important public health problem. PMID- 10793646 TI - Energy metabolism and obesity. AB - Over the long-term, most adult humans are able to maintain body energy stores through the process of energy balance, which regulates how much energy is consumed to match how much energy is expended. Energy expenditure is required for resting metabolic rate to maintain basic physiologic functions (e.g., heart beat, muscle function, respiration) and metabolize, digest, and store food that is consumed as well as for physical activity. Resting metabolic rate is the largest component of daily energy expenditure, and physical activity-related energy expenditure is the most variable. Cross-sectional studies in children and adults have shown that energy expenditure, including physical activity-related energy expenditure, are similar in lean versus obese subjects, especially after controlling for differences in body composition. A major limitation for most studies that have examined the role of energy expenditure in the cause of obesity is their cross-sectional design. Some longitudinal studies support the idea that reduced energy expenditure is a risk factor for the development of obesity, but most do not. There are several possibilities that could account for such discrepant findings. First, the ambiguous findings in the literature might be explained by the possibility that differences in energy expenditure and physical activity and their impact on the development of obesity are different at the various stages of maturation. Second, there could be individual differences in the impact of altered energy expenditure on the regulation of energy balance. The impact of energy expenditure on the cause of obesity could vary among different subgroups of the population (e.g., boys versus girls and different ethnic groups) and could have a differential effect within individuals at different stages of development. A specific example is the lower energy expenditure in Pima Indians, which predisposes to increased risk of obesity. It remains to be seen whether the lower metabolic rates that have been observed in African-Americans will relate to subsequent weight gain. It is conceivable that susceptible individuals fail to compensate for periodic fluctuations in energy expenditure. Third, given that obesity can arise as a result of a small energy balance over time, it is unlikely that existing techniques are capable of measuring such small differences. Finally, it can be argued that a focus on energy metabolism as a possible explanation of obesity is unlikely to yield interesting information because of the wide range in energy expenditure in the population even after adjusting for body composition. The major dependent variable that needs to be examined in relation to the cause of obesity is not energy expenditure but change in energy balance over time and the ability to regulate body energy stores. Given that the sudden change in obesity prevalence has occurred during a time of rapid environmental and cultural changes, additional focus on the behavioral and environmental effects on regulation of energy balance is warranted. PMID- 10793648 TI - Evaluation of the obese patient. Practical considerations. AB - The evaluation process for obesity uses the same framework of a history, physical examination and interpretation of selected laboratory and diagnostic tests that are used for other chronic care patient encounters. What makes this evaluation process different is knowing how to take an obesity focused history, what to examine, and which tests to order. An assessment of risk status is then determined based on the National Heart, Lung, and Blood Institute Clinical Guidelines on the Identification, Evaluation, and Treatment of Overweight and Obesity in Adults. Other important aspects of the evaluation process include determining the patient's goals, expectations, and motivation for weight loss along with support systems and/or barriers to behavior change. PMID- 10793647 TI - Metabolic complications of obesity. Pathophysiologic considerations. AB - Given a specific research interest in human fatty acid metabolism, this article focuses primarily on the evidence surrounding the hypothesis that dysregulation of the fuel release function of fat cells (lipolysis) is an important contributing factor to the health hazards of obesity. PMID- 10793649 TI - Dietary approaches to the treatment of obesity. AB - According to the National Institute of Health guidelines for the treatment of overweight and obesity, the most important element in a weight loss program is the reduction in energy intake. Reducing the fat content of the diet and increasing physical activity without restricting energy intake are relatively inefficient methods of weight loss. Because individuals tend to consume a constant weight of food, the decrease in energy intake on a reduced-fat diet is likely related to the lower energy density of the diet. Diets of low energy density, which are typically low in fat and rich in complex carbohydrates, allow individuals to consume satisfying portions of food while reducing their energy intake. Because a wide variety of foods can be included in a diet that is low in energy density, this type of diet encourages the adoption of life-long eating habits that are integral to the maintenance of weight loss and the prevention of weight gain. PMID- 10793650 TI - Exercise management of obesity. AB - When dealing with overweight patients, it is important to treat obesity as a chronic condition requiring long-term management. Physicians do not send a patient with hypertension home with just one bottle of pills and expect blood pressure to be managed forever. Similarly, it is naive to think that sending an overweight patient home with only a single word of advice to "lose weight" will solve the problem. Medical staff would be wise to adopt the attitude that obesity requires long-term intervention and management, just as hypertension requires ongoing monitoring and management. Continued attention to patients' physical activity is required. It is important for physicians to spend a few minutes discussing physical activity with their patients. The patient's current and past levels of physical activity should be assessed as well as the barriers to and benefits of activity. Normal-weight patients should accumulate at least 30 minutes of moderate-intensity activity on most days of the week, and overweight patients should accumulate 60 to 80 minutes per day, every day. The physician can provide some simple tools, such as a step counter and self-monitoring forms, to promote increases in activity and can follow up with the patient after the office visit. This information may sound like a lot to cover, especially given the limited time available for each patient. By spending 3 to 5 minutes intervening, however, physicians can play a critical role in promoting health. Physicians should set a goal to address one specific topic (e.g., a brief discussion of exercise benefits and barriers) and try one strategy (e.g., introduce self monitoring) during the next counseling session with patients. It is also important for physicians to remember that their success in changing behavior may be in small increments. These small changes can and do eventually lead to success for many patients and physicians. PMID- 10793651 TI - Behavioral treatment of obesity. AB - The behavioral treatment of obesity consists of a set of principles and techniques to help overweight individuals modify inappropriate eating and activity habits. As provided in University and hospital clinics, this approach produces an average loss of 8.5 kg (9% of initial weight) in approximately 20 weeks. The maintenance of weight loss is facilitated by the provision of long term patient-provider contact as well as by the use of weight-loss medications. The most promising results are likely to be obtained when behavioral and pharmacologic approaches are combined. The article concludes with a discussion of the importance of treating obese individuals with respect and compassion. PMID- 10793653 TI - Bariatric surgery. Surgery for weight control in patients with morbid obesity. AB - Morbid obesity has become a health crisis in the United States. Medical programs developed at nonoperative attempts to lose (and maintain) an adequate weight loss are largely unsuccessful. Bariatric surgery has been proven to be effective at inducing and maintaining a satisfactory weight loss to decrease weight-related comorbidity. Bariatric operations include procedures that decrease mechanically the volume capacitance of the proximal stomach (vertical banded gastroplasty, laparoscopic gastric banding) or decrease the proximal gastric capacitance and establish a partial selective malabsorption (gastric bypass and its modifications, partial biliopancreatic bypass, and duodenal switch with partial biliopancreatic bypass). These operations should induce a loss of at least 50% (or more) of excess body weight. Not all patients are candidates for these procedures, and the best results are obtained by a multidisciplinary team (including nutritionist, physician, dietitian, psychologist or psychiatrist interested in eating disorders, and surgeon). PMID- 10793652 TI - Pharmacotherapy for obesity. AB - This article discusses former and current medications used to treat obesity, reviews efficacy and safety data for current medications, and suggests clinical guidelines for their use. PMID- 10793654 TI - Public health interventions for the prevention and treatment of obesity. AB - Epidemic obesity is one of the few undesirable consequences of humanity's mastery of the environment. This article reviews public health approaches to preventing and treating obesity. The most encouraging efforts to date have been interventions targeting children in both medical and community surroundings. Treating and preventing obesity in adults has been less successful. It is suggested that taking an environmental health perspective on the obesity problem may facilitate the advancement of scientific understanding of this important health issue. PMID- 10793655 TI - Partial purification and characterization of four endodeoxyribonuclease activities from Escherichia coli K-12. AB - Four hitherto undescribed endodeoxyribonucleases, temporarily designated A(1), A(2), A(3), and B, have been isolated from E. coli K-12. Each requires Mg(++) and is not stimulated by ATP or S-adenosylmethionine. A(3) is strongly inhibited by Fe(+++) and weakly inhibited by ATP, S-adenosylmethionine, and DPN, whereas B is inhibited by caffeine. Each can be purified free of exonuclease or DNA-3' phosphatase. A(1) (molecular weight approximately 72,000) cleaves single stranded, circular fd DNA to form 3'-hydroxyl termini and introduces nicks and breaks in the closed, double-stranded replicative form DNA of fd (fd RFI). A(2) (molecular weight approximately 46,000) cleaves fd DNA and introduces nicks and breaks in RFI, forming 3'-hydroxyl- and 5'-phosphoryl termini. A(3) (molecular weight approximately 38,000) cleaves fd DNA to form 3'-hydroxyl termini and introduces only nicks in fd RFI. Irradiation of the RFI with ultraviolet light markedly increases the rate of hydrolysis by A(3). B appears to form 3' phosphoryl termini with fd DNA, but its characterization is highly preliminary due to its instability. PMID- 10793656 TI - Aliphatic analogues of nucleotides: synthesis and affinity towards nucleases. AB - DL-1-(2,3-Dihydroxypropyl)thymine was prepared by Hilbert-Johnson reaction of 2,4 dinethoxy-5-methylpyrimidine with allyl bromide followed by the osmium tetroxide catalyzed hydroxylation of the l-allyl-4-methoxy-5-methylpyrimidin-2-one obtained as an intermediate. The D-glycero enantiomer, R-1-(2,3-dihydroxypropyl)thymine and the corresponding 1-substituted uracil derivative were prepared from 3-O-p toluenesulfonyl-1, 2-O-isopropylidene-D-glycerine and sodium salt of 4-methoxy-5 methylpyrimidin-2-one or 4-methoxypyrimidin-2-one followed by treatment with hydrogen chloride in ethanol. The phosphorylation of the above 2,3 dihydroxypropyl derivatives with phosphoryl chloride in triethyl phosphate afforded the corresponding 3-phosphates which were transformed into the 2',3' cyclic phosphates by the condensation with N,N'-dicyclohexylcarbodiimide. The latter compounds of the D-glycero configuration are split by some microbial RNases to the 3-phosphates. PMID- 10793657 TI - 2'-O-methyl ribothymidine: a component of rabbit liver lysine transfer RNA. AB - One of the lysine transfer RNAs of rabbit liver is shown to contain 2'-O-methyl ribothymidine in place of ribothymidine. This represents the first demonstration of the presence of 2'-O-methyl ribothymidine in a nucleic acid. PMID- 10793659 TI - Polynucleotides. XXII. Synthesis and properties of poly 7-deazainosinic acid. AB - Poly 7-deazainosinic acid has been prepared by the deamination and phosphorylation of tubercidin and the nucleoside diphosphate was polymerised using polynucleotide phosphorylase. The polymer has similar physical properties to poly(I), but has a low thermal stability in the double-stranded complex with poly(C). Poly(7-deaza I), in contrast, forms a more stable triple-stranded complex with poly(A) than 2 poly(I). poly(A), presumably due to the higher pK value. PMID- 10793658 TI - The selective reaction of methoxyamine with cytidine residues in mammalian initiator transfer ribonucleic acid. AB - Methoxyamine reacts selectively with tRNA molecules at certain exposed cytosine residues usually located in non base-paired regions of the two dimensional clover leaf structure. Here methoxyamine is used for the first time in a study of a mammalian tRNA structure. One of the sequence abnormalities of myeloma initiator tRNA is a cytosine instead of the usual uracil immediately preceding the anticodon. A study of the reaction of the cytosine residues with methoxyamine indicates that the accessibility of bases to chemical reagents in the anticodon loop of this mammalian initiator tRNA is very similar to that observed for the bacterial initiator tRNA. PMID- 10793660 TI - Influence of template inactivators on the binding of DNA polymerase to DNA. AB - The agents daunomycin, ethidium bromide, distamycin A and cytochrome c inhibit DNA dependent DNA polymerase I (E. coli) reaction competitively to DNA. The influence of these template inactivators on the binding of DNA polymerase to native as well as denatured DNA has been determined by affinity chromatography. Cytochrome c blocks the binding of the enzyme to double-stranded and to single stranded DNA Sepharose. In contrast to these results daunomycin, ethidium bromide or distamycin A reduce the binding affinity only with denatured DNA Sepharose as matrix. These data are discussed with respect to the modification by template inactivators of the affinity of DNA to the different binding sites of the DNA polymerase. PMID- 10793661 TI - Cleavage of the glycosidic linkage of pyrimidine ribonucleosides by the bisulfite oxygen system. AB - When a solution containing 2 mM uridine, 20 mM sodium bisulfite, 0.1 mM MnCl(2), and 100 mM sodium phosphate buffer of pH 7.0 was incubated aerobically at 37 degrees or 0 degrees , partial cleavage of the glycosidic linkage of uridine took place. About 20% of the uridine was converted to uracil by the incubation for 4 hrs. Cytosine was produced from cytidine by similar treatment with bisulfite. These reactions were caused by free radicals generated by Mn(2+)-catalyzed autoxidation of bisulfite. Glycosidic bond cleavage by the bisulfite-oxygen system was not detected for adenosine, AMP, guanosine, GMP, thymidine, TMP, deoxyuridine, dCMP, dAMP, and dGMP. When poly(U) and poly(C) were treated with 20 mM sodium bisulfite in the same manner, chain fission of the polymer occurred as judged by the elution-pattern change in gel filtration through Sephadex columns. No change in the elution pattern was observed for bisulfite-treated poly(A), poly(U). poly(A) or tRNA. PMID- 10793662 TI - A new approach to the study of nucleotide sequences in DNAs. AB - The composition of the 3'terminal, 5'terminal and 5'penultimate nucleotides of the oligonucleotides released by spleen acid DNase and snail acid DNase from five ;repetitive' DNAs (guinea pig, mouse and crab satellite DNAs, yeast mitochondrial DNA and poly (dAT:dAT)) and four ;eukaryotic' DNAs (calf thymus, guinea pig and mouse liver DNAs, and yeast nuclear DNA) have been investigated and found to deviate in characteristic ways from those expected for bacterial DNAs having comparable base compositions. The deviation patterns obtained represent a novel way of characterizing and comparing different DNAs on the basis of the frequency of the nucleotide sequences they contain. PMID- 10793663 TI - The influence of subcellular fractions on the enzymic methylation of DNA in ascites cell nuclei. AB - Soluble factors appear to be present in both nuclei and cytoplasm of Krebs 2 ascites tumour cells capable of stimulating the enzyme catalysed methylation of DNA in isolated nuclei from these cells. PMID- 10793665 TI - Thermodynamic and kinetic properties of short RNA helices: the oligomer sequence AnGCUn. AB - We studied the thermodynamic, kinetic and optical properties of the double helices formed by the series of self-complementary oligonucleotides, (AP)(n)GpC(pU)(n), 2 5 x 10(3) moles/mole) while high (0.999) water activity is maintained. PMID- 10793725 TI - Correlation between three-dimensional structure and chemical reactivity of transfer RNA. AB - The bases of yeast tRNA(Phe) which react with carbodiimide and methoxyamine have been determined and this information has been combined with chemical modification studies of other workers to produce a composite picture of base accessibility in this tRNA. The results are compared with the three-dimensional structure which we have recently determined. The bases which react chemically lie in exposed positions in the three-dimensional model and those which do not are either in the double helical stem regions or else are involved in maintaining the tertiary structure through pairing or stacking interactions. PMID- 10793726 TI - Heterologous mischarging as a means of tRNA fractionation. I. Behaviour of E. coli phenylalanyl-tRNA(1Val) on BD-cellulose. AB - The chromatographic behaviour of E.coli tRNA(Val) (1), Val-tRNA(Val) (1) and Phe tRNA(Val) (1) was studied on BD-cellulose columns. At pH 4.0 and 4 degrees C the elution position of Phe-tRNA(Val) (1) was not affected by the presence of absence of Mg(2+), whereas Val-tRNA(Val) (1) was slightly retarded when Mg(2+) was ommited. It is postulated that the amino acid and its nature influence the structure that the aminoacyl-tRNA assumes. Under suitable conditions the heteroaminoacylated Phe-tRNA(Val) (1) eluted significantly later than other tRNAs. This fact showed that heterologous mischarging can be a useful step in tRNA purification methods. PMID- 10793727 TI - Urea perturbation and the reversibility of nucleohistone conformation. AB - Urea effect on conformation and thermal stabilities in nucleohistone and NaCl treated partially dehistonized nucleohistones has been studied by circular dichroism (CD) and thermal denaturation. Urea imposes a CD change at 278mm of DNA base pairs in native and NaCl-treated nucleohistones which can be decomposed into two parts: a decrease in Deltaepsilon(278) for histone-free base pairs and an increase for histone-bound base pairs. The reduction by urea of Deltaepsilon(220) of bound histones is approximately proportional to the increase of Deltaepsilon(278) of histone-bound base pairs. Urea also lowers the melting temperatures of base pairs both free and bound by histones. The presence of urea indeed destroys the secondary structure of bound histones, causing changes in the conformation and thermal stabilities of histone-bound base pairs in nucleohistone. Such a urea perturbation on nucleohistone conformation is reversible. PMID- 10793728 TI - The thermal denaturation of DNA: average length and composition of denatured areas. AB - A spectral study of melting curves of DNA ranging from 73 to 32% AT indicates that the base ratio of sequences melting within DNA are a linear function of temperature. A study of partially denatured DNA by electron microscopy, reversible renaturation and fractionation on hydroxylapatite suggests that the melting curve of DNA represents the melting of sequences which average 3-4 million daltons in length. These sequences appear to be a combination of two areas, one which is high in AT and denatures in the first three-quarters of the melting curve, and one which is high in GC and denatures in the final quarter. The length of these sequences appears to vary between 1.5-6 million daltons. PMID- 10793729 TI - The preparation and characterisation of chromatin from third instar larvae of Drosophila melanogaster. AB - A procedure is described for the isolation of large amounts of chromatin from Drosophila third instar larvae. This material is suitable for immunisation of rabbits, and preliminary analysis by immunodiffusion reveals little cross reaction between chromatin and cytoplasm antigens, using antisera prepared against them. In addition to these studies, results from an examination of the exchange of radioactively labelled cytoplasmic proteins with nuclei suggest that no more than 5% of the total chromatin protein is of cytoplasmic origin. Electrophoretic analysis indicates that of this 5%, 20% migrates as a single band, in the same position as f2b histone. The techniques of radioactively labelling larvae which were used, may be suitable for achieving specific activities of up to 100muC/mg larval protein. Contamination of the chromatin by yeast proteins from the larval food is no more than 10%. Of this 30% is represented by one electrophoretic band. Immunological analysis reveals little cross reaction with yeast extracts. Electrophoretic and amino acid analyses of the chromatin proteins confirm earlier reports concerning Drosophila histones. PMID- 10793730 TI - Proton magnetic resonance studies of ultraviolet-irradiated apurinic acid. AB - In apurinic acid, a single-stranded polydeoxyribonucleotide easily obtained upon depurination of DNA, the proton resonances arising from thymine and cytosine are readily observable in aqueous solution of 25 degrees C. Two methyl thymine resonances, centered at 1.88 ppm and separated by 0.045 ppm, are observed. We attribute the downfield methyl resonance to thymines with no pyrimidine nearest neighbors and the upfield methyl resonance to thymines having pyrimidine neighbors in the 3' and/or 5' positions. Upon ultraviolet irradiation, the upfield methyl and thymine H-6 resonances decrease in amplitude and two methyl resoances appear at 1.63 and 1.52 ppm, corresponding, respectively, to cytosine thymine and thymine-thymine cyclobutane dimers. Photoreversal eliminates these two minor methyl resonances from the pmr spectrum. We conclude that apurinic acid provides a suitable model system for pmr studies of chemically modified pyrimidine bases in DNA. PMID- 10793731 TI - The fractionation of t-RNA on N,N'-bis(3-aminopropyl)-piperazine substituted Sepharose. AB - An anion exchange agarose has been prepared by modifying sepharose 6B with N,N' bis (-3-aminopropyl) piperazine. This material (BAPP-Sepharose) has been used for the fractionation of t-RNA from E.coli by column chromatography. The results obtained with gram quantities of crude t-RNA at pH 4.6 and pH 8.0 as measured by the elution patterns of alanyl, arginyl, aspartyl, leucyl, lysyl, methionyl, phenylalanyl, prolyl, seryl, tyrosyl, and valyl t-RNA are described. PMID- 10793732 TI - CD studies on the conformation of some deoxyoligonucleotides containing adenine and thymine residues. AB - CD studies of the deoxyoligomer series d(pT)(n) and d(pA)(n) show increasing CD maxima for oligo (dT)'s with chain length variation from two to seven, while oligo (dA)'s exhibit a decreasing CD maximum. Concentrated solutions of NaClO(4) cause a decrease in the CD of longer oligo (dT)'s towards the CD of d(pT)(2) which is different from oligo dA's. Probably base-sugar interactions are important in the observed conformational effects. The chemically synthesized oligomers dpApApTpT and dpTpApTpA show deviations in their CD spectra which reflect a dominating conformational effect of d(pA)(2) in the former but not in the alternating isomer. PMID- 10793733 TI - Oligonucleotide conformations. 4. Temperature and solvent effects on different uridylyl-uridines. PMID- 10793734 TI - The premelting of nucleoprotein: role of non-histone proteins. AB - In native nucleoprotein, the premelting structural changes of DNA are not observed by circular dichroism measurements. In order to determine which protein fraction of chromatin is responsible for the absence of premelting we have examined a series of nucleoproteins depleted of different protein fractions by treatment with sodium chloride or sodium deoxycholate.The premelting reappears as soon as non-histone proteins are removed or in residual complexes from which the two slightly lysine-rich histone fractions (F2a2+F2b) have been removed. On the other hand, it is shown that histone F1 alone is not able to suppress the premelting phenomenon. It is thus concluded that the absence of premelting is a property of native nucleoprotein where interactions between the different proteins complexed with DNA can occur and especially between the non-histone proteins and the two slightly lysine-rich histone fractions. PMID- 10793736 TI - Different deoxyribonucleases in human lymphocytes. AB - The distribution pattern of deoxyribonuclease activities in human lymphocytes has been examined by micro-disc-electrophoresis. Four groups of deoxyribonuclease activities, differing in their electrophoretic mobility, in the nature of their optimal substrate and in their optimal incubation conditions, are characterized. There are two alkaline DNase-activities. One corresponds to DNase I (EC 3.1.4.5), the other having pH optimum of about pH 9.0, prefers denatured DNA as substrate and is not dependent on divalent cations. The fractions with an acid pH optimum can be subdivided into two groups, which differ in their activity towards native DNA, towards denatured DNA, in their activity when succinate is present and in their pH optimum. PMID- 10793735 TI - Deoxynucleotide substitution: a new technique for sequence analysis of RNA. AB - It is possible to replace in a normal transcription reaction catalyzed by E.coli RNA polymerase one of the four precursors by the corresponding deoxynucleoside triphosphate. These deoxynucleotide-substituted RNA's offer interesting prospects for nucleotide sequence analysis. Indeed by the use of U(2)-RNase with dG-RNA, or pancreatic RNase with dC-RNA or dU-RNA, base specific cleavage can be obtained at any of the four residues. In this way overlap of at least six residues in length can be obtained for any site in the RNA. The technique offers also great benefit for solving the sequence of the more difficult T(1)-oligonucleotides. Some examples in the sequence analysis of SV40 DNA-Hind fragments are reported. PMID- 10793737 TI - Terminal oxidation-reduction of N-acetyl-phenylalanyl-tRNA blocks initiation complex formation with Escherichia coli 30S ribosomal subunits. AB - Terminally oxidized-reduced tRNA(Phe) of yeast, exclusively acylated at the 2' hydroxyl of the 3'-terminal ribose of the tRNA, is a useful model for investigating the stereospecificity of AA-tRNA in protein synthesis. In this work, the ability of N-acetyl-Phe-tRNA(ox-red) to form an initiation complex with Escherichiacoli 30S ribosomal subunits was investigated. Thirty per cent of added control N-acetyl-Phe-tRNA was bound in a reaction dependent on initiation factors, GTP, and poly U, but no binding of the oxidized-reduced analog could be detected. These results imply that initiation complex formation may be specific for the 3'-ester of initiator tRNA. PMID- 10793738 TI - Synthesis of 1,N6-etheno-2-aza-adenosine (2-aza-epsilon-adenosine): a new cytotoxic fluorescent nucleoside. AB - 1,N(6)-Etheno-2-aza-adenosine was synthesized by treating 1,N(6)-etheno-adenosine with alkali, followed by nitrosation. The mechanism of formation of this novel nucleoside was elucidated using adenosine tritiated at C-8 and C-2, and was found to deformylate exclusively at C-2. This new 2-aza nucleoside fluoresces at 494 nm when excited at 358 nm. Toxicity study showed the compound is active in a rat mammary tumor tissue culture line, but inactive in HeLa and Glioma 26 tissue culture lines. It was also found to selectively inhibit the thymidine incorporation into DNA in a rat mammary tumor, but exhibits no ill effect on normal proliferative tissue. The reactive intermediate 3-beta-D-ribofuranosyl-4 amino-5-(imidazol-2-yl) imidazole was identified and was found to be an active agent in tissue culture. PMID- 10793739 TI - Unpaired bases in superhelical DNA: kinetic evidence. AB - Kinetic analysis of the early, fast reaction of superhelical DNA with formaldehyde reveals that this region or regions is 56% "single strand like" in character. Hydrogen-tritium exchange studies coupled with other considerations show that this reaction is not due to a difference in conformational motility between form I and form II molecules, but is due to unpaired or weakly hydrogen bonded, localized region(s) of the form I allomorph of circular DNA. PMID- 10793740 TI - Fractionation and characterization of satellite DNAs of the kangaroo rat (Dipodomys ordii). AB - Nuclear DNA from liver cells of the kangaroo rat species Dipodomysordii was fractionated and characterized with the aid of buoyant density gradients in neutral and alkaline CsCl and in Ag(+)-Cs(2)SO(4). More than one-half of the DNA was present in three density satellites, a greater proportion than in any other species yet reported; the purified satellite DNAs were denser than principal DNA. All satellite fractions revealed sharp isopycnic bands and narrow denaturation profiles. Two had identical buoyant densities but differed substantially in T(m), base composition, and reassociation kinetics. In alkaline CsCl all three satellites, as well as a shoulder of intermediate repetitive DNA on the heavy side of the principal band, revealed unique strand densities. The most highly repetitive satellite was unusually rich in (G + C) and contained 6.7% of 5 methylcytosine. A survey of internal organs and spermatozoa of an adult male revealed no significant differences in distribution of the satellites among tissues. PMID- 10793741 TI - The nucleotide sequence preceding an RNA polymerase initiation site on SV40 DNA. Part 1. The sequence of the late strand transcript. AB - The nucleotide sequence of the transcript of the "late" strand of the region of SV40 DNA preceding the preferred initiation site for Escherichia coli RNA polymerase has been determined to be U-G-U-A-A-C-C-A-U-U-A-U-A-A-G-C-U-G-C-A-A-U A-A-A-C-A-A-G-U-U-A-A-C-A-A-C-A-A-C-A-A-U-U-G-Cp. Hemophilus influenza restriction endonuclease cleaves this region 30 nucleotides (base pairs) before the site of initiation of RNA synthesis by RNA polymerase. PMID- 10793742 TI - The nucleotide sequence preceding an RNA polymerase initiation site on SV40 DNA. Part 2. The sequence of the early strand transcript. AB - The nucleotide sequence of the RNA transcript from the "early" (E) strand of SV40 DNA immediately preceding the preferred E. coli RNA polymerase start site is G-(A A-A-C, -A-U-)-A-A-A-A-U-G-A-A-U-G-C-A-A-U-U-G-U-U-G-U-U-G-U-U-A-A-C-U-U-G-U-U-U-A U-U-G-C-A-G-C-U-U-A-U-A-A-U-G-G-U-U-A-C-Ap. The last nucleotide of the sequence is the first nucleotide transcribed by E. coli RNA polymerase from the "E" strand. The DNA template contains a palindrome of 17 residues that includes the Hemophilus influenza restriction endonuclease cleavage site G-T-T-A-A-Cp. The DNA which gives this transcript lies very close to one end of SV40 DNA segment in the Adeno-SV40 hybrid virus Ad2+ND3 and appears to contain sufficient untranscribed information to specify the E. coli RNA polymerase start. PMID- 10793743 TI - Use of phosphorus oxychloride in synthesizing nucleotides and oligonucleotides. AB - Procedures are described for phosphorylating protected nucleotides, oligonucleotides and phosphoramidate oligonucleotide derivatives at the 3' hydroxyl group. The conditions (phosphorylation with phosphorus oxychloride and pyridine in dioxane followed by hydrolysis with aqueous pyridine) are sufficiently mild that base labile (trifluoroacetylamino; beta-cyanoethyl phosphotriester) and acid labile (O-monomethoxytrityl; phosphoramidate) functions are retained intact. Application of the technique is illustrated by the synthesis of dpT, dTp, d(CF(3)CONH)Tp, dTp(N)Tp, and dTp(N)Tp(N)Tp. In addition, the utilization of phosphorus oxychloride in joining thymidine derivatives and dinucleoside phosphotriester blocks via phosphodiester links is described. PMID- 10793744 TI - Transport of nucleosides in Bacillus subtilis: characteristics of cytidine uptake. AB - In Bacillus subtilis SMYW cytidine and uridine are transported by a common system. Transport of these substances requires metabolic energy. After 60 sec of (3)II-cytidine-uptake practically all accumulated radioactivity was found in phosphorylated products. Addition of ribonucleosides with inhibitory effect upon cytidine-uptake resulted in competitive type of inhibition while interference with deoxyribonucleosides was of hyperbolic competitive type. Adenosine possesses a high affinity to cytidine-transporting site but requires different system(s) for its own transport. Guanosine, adenine, cytosine and 5-nucleotides do not interfere with cytidine-uptake. PMID- 10793745 TI - Transport of nucleosides in Bacillus subtilis: the effect of purine nucleosides on the cytidine-uptake. AB - The structural effects of chemical modifications upon the affinity of purine nucleosides to cytidine-transport system in Bacillus subtilis were investigated using a series of modified derivatives. The interaction involves protein molecule(s) which require the presence and proper orientation of the sugar residue and its hydroxylic functions. Moreover, a specific interaction with the heterocyclic ring system is involved in the process which results in a requirement for an aromatic pi -electron system and an absence of a polarizable function at position 6 of the purine heterocycle. The region in the protein responsible for the latter interaction is rather limited and, consequently, a proper nucleoside conformation is required. PMID- 10793746 TI - Inducible and constitutive nucleoside-binding sites in Escherichia coli: differential inhibition by nucleoside analogues. AB - Constitutive and inducible nucleoside-binding sites on the surface of the cells of E. coli B have different patterns of inhibition by nucleoside analogues. 1(beta-D-Ribofuranosyl)-4-aminopyrimidine-6-one and isoguanosine preferentially inhibit the inducible binding sites while showdomycin and N(4)-dimethylcytidine interfere more strongly with the constitutive function. PMID- 10793747 TI - Isolation and sequence determination of the 3'-terminal regions of isotopically labelled RNA molecules. AB - The method which was developed for the selective isolation of 3'-terminal polynucleotides from large RNA molecules on columns of cellulose derivatives containing covalently bound dihydroxyboryl groups has been modified and adapted for use on radioactively labelled RNAs. The 3'-terminal polynucleotide fragments which result from specific ribonuclease digestion of isotopically detectable quantities of RNA can be selectively obtained in both high yield and purity by the modified procedure and can be subsequently analyzed by standard electrophoretic and chromatographic techniques. In addition, when the extent of enzymatic fragmentation of the RNA is controlled, the procedure permits the selective isolation of discrete "sets" of fragments of variable chain length, all of which derive from the 3'-terminus of the RNA molecule. These overlapping polynucleotides can be used directly to obtain extensive sequence information regarding the primary structure in the 3'-region of the RNA. PMID- 10793748 TI - Mercury(II) binding to s4U in E. coli tRNA(Val). AB - The accessibility of the s(4)U base in native tRNA(Val) from E.coli was monitored by studying the binding of various mercurials. The relative binding order HgBr(2)[unk]HgCl(2)>>CH(3)HgOAc[unk]CH(3)HgCl[unk]PCMB parallels approximately the steric requirements of linear HgX(2) or RHgX compounds for S(N)2 displacement by sulfur, although other factors are operative. Para-chloromercuri-benzoate (PCMB) does not bind the thiolated nucleotide unless the tertiary structure of the tRNA is opened up by removal of Mg(2+) ions and heating to 40 degrees . Under these conditions, equilibrium dialysis measurements using (14)C-labeled PCMB showed one binding site (n = 0.93) with an association constant, K(1), of 9 x 10(4)M(-1). PMID- 10793749 TI - Degradation of 6-methyl-2'-deoxyuridine in the in vivo and in vitro systems of Escherichia coli. PMID- 10793750 TI - Mung bean nuclease: mode of action and specificity vs synthetic esters of 3' nucleotides. AB - Mung bean nuclease hydrolyzes synthetic esters of 3'-nucleotides to nucleosides and phosphate esters; esters of 2'-nucleotides, and 2'--> 5' internucleotide linkages, are resistant. Esters of ribonucleotides are cleaved at 100-fold the rate for deoxyribonucleotides, the increased rate being due to presence of the 2' hydroxyl and not to differences in conformation. Introduction of a 5'-substituent leads to a 3-fold increase in rate. The rates of hydrolysis vary up to 10-fold with the nature of the base, in the order adenine > hypoxanthine > uracil; and up to 6-fold with the nature of the ester radical. This form of cleavage of esters of 3'-nucleotides is also characteristic for nuclease-3'-nucleotidase activities from potato tubers and wheat, suggesting that one type of enzyme is responsible for all these activities. PMID- 10793751 TI - Transfer RNA methylating activity of yeast mitochondria. AB - Mitochondria isolated from Saccharomyces cerevisiae and purified in Urografin or sucrose gradient contain tRNA methylating activity with specificities different from those of the cytoplasm. The main reaction product, using E.coli tRNA as methyl group acceptor, is N(2),-N(2)-dimethylguanine. The corresponding mitochondrial methylase is coded by nuclear DNA. A DNA methylating activity is also associated with yeast mitochondria. PMID- 10793752 TI - Effect of L-methioninyl adenylate on the level of aminoacylation in vivo of tRNA(Met) from Escherichia coli K12. AB - In cells of E.coli K12 grown exponentially in minimal medium, tRNA(met), tRNA(leu) and tRNA(ile) are aminoacylated at 100%, 80% and 64%, respectively. On addition of L-methioninyl adenylate to the growth medium, one observes a specific deacylation of tRNA(met). When more than 35% of tRNA(met) is deacylated, growth rate is reduced and becomes proportional to the amount of methionyl-tRNA formed. PMID- 10793753 TI - The mapping and ordering of fragments of SV40 DNA produced by restriction endonucleases. AB - SV40 DNA is cleaved by the Eco RII and Hae restriction endonucleases to give rise to two different sets of 16 fragments each. These fragments have been ordered by analysis of the products of redigestion of one set of fragments with another restriction enzyme. The cleavage sites have been precisely mapped based on length measurements of the products of each cleavage. This provides a convenient group of small DNA fragments suitable for sequence analysis investigation of the transcripts present in infected cells, or construction of deletion substitution variants of the virus. PMID- 10793754 TI - 'DNA snapback' peptides. AB - Thermal denaturation studies show that 10-15% of the calf thymus DNA in the heat denatured (Tyr-Gly-Tyr-Gly-Tyr)-DNA complex renatures spontaneously after colling. The double-strandness of this DNA was verified by its resistance to single-strand Neurospora endonuclease and by its elution profile on hydroxypatite columns. The renatured DNA isolated by the latter technique was found to contain 56% GC compared to the 41% GC content of the whole thymus DNA. Alternating tryptophanyl-glycyl and histidyl-glycyl peptides also catalyze the same renaturation. A linear correlation was found between the thermal stabilization afforded to the DNA by the various peptides and their ability to "catalyze" DNA strand renaturation. PMID- 10793755 TI - Interaction of tyrosyl, histidyl, and tryptophanyl peptides with DNA: specificity and mechanism of the interaction. AB - In the Tyr-(Gly)(1-4)-Tyr series maximal thermal stabilization of calf thymus DNA (deltaT(m)=10 degrees ) occurred with the Tyr-(Gly)(2)-Tyr peptide, where three base pairs could separate the two tyrosyl residues. Tyr-Gly-Tyr-Gly-Tyr stabilized the DNA by 6 degrees . The alternating Trp-Gly-Trp-Gly-Trp and His-Gly His-Gly-His peptides were equally as effective as the Tyr-Gly-Tyr-Gly-Tyr peptide in stabilizing calf thymus DNA against thermal denaturation. But the alternating Phe-Gly-Phe-Gly-Phe peptide afforded little stabilization, suggesting that a sidechain possessing both a conjugated pi-electron system and an electron donor atom is necessary for DNA stabilization. Introduction of electron withdrawing iodo or nitro group into the tyrosyl sidechains almost completely abolished the stabilizing effect. Although the tyrosyl peptides seem to be specific for GC-base pairs, no correlation was found in natural DNA between% GC and% thermal stabilization. Eukaryotic DNAs showed twice the stabilization of prokaryotic DNAs with the same GC content. PMID- 10793756 TI - New fluorescent hydrazide reagents for the oxidized 3'-terminus of RNA. AB - The synthesis and properties of four new fluorescent reagents capable of forming moderately stable links to the 3' oxidized end of RNA are reported. All are hydrazide derivatives: pyrene butyric acid hydrazide, proflavine monosemicarbazide, proflavine monosuccinic acid hydrazide, and anthracene-9 carboxaldehyde carbohydrazone. In addition, procedures are given for coupling the bifunctional reagent carbohydrazide to the 3' end of RNA. These carbohydrazide adducts can easily be coupled in turn to a wide variety of fluorescent reagents having specificity for aliphatic amino groups, including isothiocyanates and sulfonyl halides. Thus a route exists for the preparation of an enormous variety of 3' fluorescent labeled RNAs. The carbohyrazide adducts are also useful for other synthetic procedures such as preparation of covalent tRNA dimers. PMID- 10793757 TI - Hybrid isolation by recovery of RNA-DNA hybrids from agar using S1 nuclease. AB - A method for recovering RNA-DNA hybrids from agar employing a single strand specific nuclease is described. The procedure is suitable for large scale isolations, and immobilization of the DNA in agar prior to hybridization allows a high yield of hybrid without interference by DNA reannealing. PMID- 10793758 TI - Nucleoside conformations. 15. Flexibility of natural pyrimidine nucleosides around the glycosidic bond. AB - The flexibility of pyrimidine nucleosides has been investigated by measuring their circular dichroism in hydroalcoholic solutions over a large temperature range (-100 degrees to +40 degrees C). It was observed that ss-Uridine (Urd) and ss-Cytidine (Cyd) showed a decrease of the main dichroic band of about 40 percent, while sterically hindered nucleosides (alphaUrd, ara-Uracile, 02-2' anhy dro-Urd, 2'3'-0-isopropylidene-Urd) showed only small decreases. It is concluded that the flexibility of the glycosidic linkage in conjunction with the pseudo rotation of the sugar residue is responsible for these changes; the thermodynamic values which can be deduced from these decreases are compatible with an oscillatory motion around the glycosidic bond, but exclude anti-syn transformations in pyrimidine nucleosides. PMID- 10793759 TI - "Methylation-coupled" transcription by virus-associated transcriptase of cytoplasmic polyhedrosis virus containing double-stranded RNA. AB - S-adenosyl-L-methionine (SAM) activated the virus-associated RNA polymerase of cytoplasmic polyhedrosis virus in vitro. Synthesis of single-stranded viral RNA (mRNA) proceeded depending on the presence of SAM.A methyl residue of SAM was incorporated into an RNA molecule. A ribose moiety of adenylic acid in the 5' terminal region of the nascent RNA was methylated in the very early stage of the transcription. The dependence of the viral transcription on the presence of SAM and the methylation of terminal nucleotide suggests that the transcription of CPV is a "methylation-coupled" reaction. PMID- 10793760 TI - A conformational study of some adenosines by use of nuclear Overhauser effect. AB - Conformations of 8-bromo-2'-[unk]-triisopropylbenzenesulfonyladenosine ([unk]) and its 3'-[unk]-isomer ([unk]) in solution have been determined by the use of intramolecular nuclear Overhauser effects in (1)H NMR spectroscopy. Compound [unk] has been proved to have a conformation in which the adenosine and benzene rings are intramolecularly stacked and compound [unk] an elongated non-stacked conformation in dimethylsulphoxide. The 5'-[unk]-acetyl derivative of [unk] has also been found to adopt the intramolecularly stacked conformation in dimethylsulphoxide, but a non-stacked one in chloroform. Coupling constants observed are discussed in connection with the conformation of the ribose moiety. The (13)C NMR spectra have also been examined, but no effect which could be ascribed to the stacking phenomena was observed in the carbon chemical shifts. PMID- 10793761 TI - Protein synthesis of the sponge Geodia cydonium: characterization of the system. AB - The ribosomal population of the sponge Geodia cydonium has been examined. The monosomes have a sedimentation constant of 80 S, the sizes of the subunits are approximately 60 S and 45 S respectively. The polyribosomes contain up to 40 ribosomal units. Cell free protein synthesizing systems (cell homogenate as well as reconstituted system) have been prepared and characterized with respect to Mg(2+), KCI and ATP concentrations, temperature, pH and time course of the reaction. In the cell-free system and in the cellular system the protein biosynthesis is inhibited by chloramphenicol. It is not affected by cycloheximide. PMID- 10793762 TI - Non ductal-adenocarcinoma neoplasms of the pancreas. AB - Pancreatic Non Ductal-Adenocarcinoma Neoplasms (PNDAN) represent about 20% of pancreatic and periampullary tumors and should be considered in differential diagnosis with ductal adenocarcinoma in the presence of isolated pancreatic mass. From January 1992 to December 1998, 238 patients were operated on for pancreatic and periampullary masses. Fifty-five patients had PNDAN: 24 endocrine tumors, 7 serous cystadenomas, 6 intraductal papillary-mucinous tumors, 5 acinar carcinomas, 4 mucinous cystadenomas, 3 metastatic tumors, 2 cystic papillary tumors, 2 solid cystadenocarcinomas, 1 neurilemmoma, and 1 pancreatoblastoma; 19 were benign and 36 were malignant or borderline tumors. A correct preoperative diagnosis was obtained in 58% of the cases. In all other cases, diagnosis was achieved intraoperatively. Major (18 pancreaticoduodenectomies, 17 left splenopancreatectomies, 1 total pancreatectomy) and minor resections (5 central pancreatectomy, 10 enucleations) were performed; curative surgical operations were carried out on 39/55 patients (curative resectability: 71%). Operative mortality and morbidity were 1.8% and 21.8%, respectively. Three and 5-year actuarial survival for malignant or borderline PNDANs are 65% and 40% versus 31% (3-year) for ductal adenocarcinoma of pancreatic head treated by pancreaticoduodenectomy (p-value = 0.03). We believe that pancreatic masses that are not ductal adenocarcinomas, can be aggressively resected even if large in size, resulting in a better outcome than ductal adenocarcinoma itself. PMID- 10793763 TI - Second primary lung cancer: possible prediction, treatment and prognosis. AB - The incidence of second primary lung cancer (SPLC) seems to have increased in recent years. In our series that included more than 4000 patients with lung cancer and 1600 operations, we observed 26 cases of SPLC up until June 1999. We considered the standard criteria accepted by most authors for the selection. Characteristics of patients and tumors at the time of the first operation were compared with the statistical data of our general series. Age, sex, performance status and smoking were all considered, as well as cancer site, histology, stage and type of first surgical operation. No significant independent factor could be identified for the prediction of a new tumor at that time. A second operation was possible in 16 cases: two patients died in the postoperative period, two survived for 12 and 24 months, respectively, 12 are still alive 8-87 months after the new resection. The high rate of surgical cases (16/26 = 62%) and a good survival rate is clearly correlated with an early diagnosis of SPLC. PMID- 10793764 TI - [Carcinoma of the hypopharynx: reconstruction of the pharyngo-esophageal tract after circular pharyngectomy by the transplantation of a U-shaped jejunal loop to the neck]. AB - Reconstruction of the oro- and hypopharynx has specific difficulties duo to their wide diameters. Thirteen patients underwent reconstruction with a free U-shaped jejunal transplant, after circular pharyngo-laryngectomy for hypopharyngeal cancer invading the oropharynx. This transplant included a side-to-side anastomosis between the two limbs of the jejunal loop and allowed reconstruction of the upper digestive tract after wide carcinologic resection of the pharynx. The U-shaped jejunal transplant facilitated the upper anastomosis, especially the upper part where the resection involved the oropharynx. It formed a reservoir behind the tongue and avoided nasal reflux. Best indication are large resections involving the oropharynx. PMID- 10793765 TI - [Surgical treatment of complicated diverticular disease of the colon]. AB - The present study analyzes the results obtained by the AA with the different types of surgery adopted in the treatment of the complicated diverticulosis of the colon, highlighting, on the basis of data available in literature, the possible treatments in the different clinical settings. A retrospective study analyzing type of complication, the surgical technique adopted, Hinchey stage, mortality and morbidity rates and average hospital stay correlated with the kind of intervention has been carried out on 83 surgical interventions performed between 1984 and 1988. The results show that 43 R.A.P. (R.A.P. = primitive anastomosis resection) (32 cases at the I-II stage and 11 cases at the III-IV stage), 27 Hartmann (11 at the I-II and 16 at the III-IV), 9 colostomies (2 at the I-II and 7 at the III-IV), 2 esteriorizations and 2 simple drains have been carried out on a total of 44 intestinal perforations, 16 recurrent diverticulitis, 13 intestinal occlusions, 2 fistulae, 5 abscesses and 3 hemorrhages. The total mortality rate amounts to 10.6%; the morbidity rate of the R.A.P. interventions to 14.4 (I-II stage-related morbidity = 15.6%, III-IV stage = 63.6%), Hartmann's to 9.6% and that of the colostomies to 3.6%. Furthermore, in this work, we have considered the cases of riconversation after Hartmann interventions (9 cases): in the second operations the mortality and morbility rate amounts to 0 and the hospital stay to 9 days. The AA analyze on the surgical technique adopted in the different cases and the of choice criteria. According to the data obtained and to current literature, it results that the primitive anastomosis resection represents the first choice intervention at the I-II stage, although, in selected cases, it can be carried out also at the III-IV stage. Hartmann surgery confirms its effectiveness while simple colostomy is no longer accepted in literature. PMID- 10793766 TI - [Asymptomatic cholelithiasis: indications for cholecystectomy based on the levels of acute phase proteins]. AB - Prophylactic cholecystectomy for asymptomatic gallstones is still controversial. Aim of the study was to assess whether the determination of serum acute phase proteins (APP) could be utilized as a criterion for cholecystectomy, as they are suggestive of the presence in the blood of cytokines released from the inflamed gallbladder wall, even when clinical signs are missing. In 75 cases of gallstones, free from other coexistent inflammatory processes, red cell sedimentation rate, plasmatic cortisol, immunoglobulin (IgA, IgG, IgM), electrophoresis of the proteins, CPR, fibrinogen, haptoglobin, alfa-1-antitrypsin and bile culture have been detected. The patients have been subdivided into two groups: patients with asymptomatic or mildly symptomatic disease and patients with a clear clinical pattern of acute cholecystitis. In the latter alfa-1 globulin, alfa-2-globulin, ESR, CPR and cortisol turned out to be significantly elevated, while in 20-30% of the former CPR, beta-globulin and cortisol were increased, too. The study demonstrates that among the patients with asymptomatic gallstones there is a population having PFA values higher than normal. This is suggestive of a cytokines activation which, when other inflammatory processes can be excluded, is likely due to gallbladder inflammation and surgery will likely be indicated. PMID- 10793767 TI - [Th1-Th2 cytokine correlates (INF-gamma and IL-4) in splenectomized patients]. AB - This paper attempts to explain if immunodepression in patients who had undergone a splenectomy may be due to altered balance between Th1-Th2 lymphocyte subpopulations, as shown in several studies on phagocyte and lymphocyte cells. This was achieved by dosing serum levels of IFNg, produced by Th1 lymphocytes and IL-4, produced by Th2 lymphocytes. Final analysis showed immunodepression in splenectomized patients but also emphasized that in 70% of all cases, there is functional damage of T-lymphocytes that continues for several years after the surgery involving both cellular and humoral immunity. Immunoglobulin dosage allows the increase of IgE to be seen in 50% of the splenectomized patients studied, all with allergic symptoms that appeared after the operation. The production of IgE is stimulated by Th2 lymphocytes. This leaves one to believe that splenectomy may favour the persistence of allergens in the blood, the appearance of allergic symptoms and the increase of IgE serum levels in patients with normal Th2 functioning and consequently, with normal or increased IL-4 serum levels. PMID- 10793768 TI - [A probiotic as an antagonist of bacterial translocation in experimental pancreatitis]. AB - Infection is the most common cause of death in acute pancreatitis. Earlier studies have demonstrated that early enteral nutrition decreases microbial translocation, upregulates the immune function and reduces septic complications and mortality. Lactobacillus plantarum (Lp) has been shown to be effective in reducing egress of endotoxin and microbial strain that showed very high adherence power to gut mucosa. We adopted a model of acute pancreatitis induced by isolation and ligation of biliopancreatic duct in adult Lewis rats. Three groups were studied: A. control group (sham operation); B. induced pancreatitis, no further treatment; C. Induced pancreatitis + gavage with 5 ml/day of a suspension of Lp 299 v in a dose of 0.5-1.0 x 10(9)/ml during 4 days before and 4 days after induction of pancreatitis. All animals were sacrificed after 96 hours. Histological studies and microbiological analyses were performed. Forty out of 55 animals showed signs of severe pancreatitis on sacrifice after 96 hours. Only these animals were further studied. In group A, we found only 1/20 bacteria in mesenteric nodes (MN). Pathogenic microrganisms were found in the non-treated group in MN in 14/20 and in the pancreatic tissue in 10/20. In contrast, when kept on an umbrella of Lp 299 v, only 4/20 animals demonstrated growth of enteric bacteria in MN and 3/20 in pancreatic tissue. All of these results showed a significant reduction of infection in the treated groups. In our model, Lp 299 v is effective in preventing microbial translocation in experimental pancreatitis. Treatment with probiotic bacteria, such as Lactobacillus spp, seems to be a promising alternative as problems with antibiotic-resistant bacteria seem to accumulate. PMID- 10793769 TI - [Towards a new classification of hemorrhoidal disease]. AB - The generally accepted classification of hemorrhoids presents some rather wide gaps which may either be due to expressions of intrinsic limits or recent moves of the Colonproctology field. The authors of this paper would like to present their proposal for a new classification that considers the results of the National Multicentric Study that was performed in collaboration with nineteen centres and a total of 1,494 patients. In addition, a comparative study was conducted on the two forms of classification. Nineteen patients (1.28%) were considered unclassifiable with the new classification system while 49 were (3.29%) with the presently used one. These results were found significant with the MacNemar Chi-Square test. The description of type 2 and 3 in the new classification system was found useful in 69% of 2nd degree, 77% of 3rd degree and 28% of 4th degree using the present system. Acute events (type 5,6 and 7) proposed with the new classification were found in the present system with several different rates. We have also included the data taken from a national and international survey we conducted on the usefulness of the new classification that provoked intense debate within two of the most important national institutes. The result was the elaboration of a design on the new classification. PMID- 10793770 TI - [Effect of octreotide on the hepatic metastasis of human colorectal cancer: an experimental model in athymic mice]. AB - The aim of this randomized study was to determine the effects of octreotide therapy on the growth and development of experimental liver metastases from a human colonic cancer cell line (HT 29) in nude mice model. No important and significant difference could be found between mice, lungs and liver weights of both groups as well as lung metastatization; indeed, significant was the difference between groups concerning liver, metastases (the majority of them were in treated group): in spite of the small number of data collected, which does not allow to draw any conclusion on the efficacy of this drug on liver metastases, we believe that octreotide therapy does not affect dramatically the growth and development of liver metastases from a human colon cancer. PMID- 10793771 TI - [A case of intestinal occlusion caused by endometriosis of the cecum]. AB - The purpose of this study is to heighten awareness of intestinal endometriosis, a disease that may mimic other abdominal pathologies (bowel carcinoma, intestinal inflammatory disease, diverticulitis), sometimes found in the emergency setting. The Authors report a case of acute bowel obstruction due to coecal endometriosis with appendix mucocele, peritoneal pseudomyxoma and ovarian endometrioma. The patient was operated on in the emergency setting, a right colectomy was performed and she then received pharmacological suppressive treatment with Gn-RH analogues and danatrol. We remark that preoperative diagnosis is very difficult in those cases that do not have a past history of pelvic endometriosis. An accurate anamnesis regarding the chronology of pain onset (typically only during the menstruation at first), but especially intraoperative histopathologic examination are useful for diagnosis. An increased awareness of intestinal endometriosis in reproductive age women with acute bowel obstruction, associated with an accurate anamnesis of menstrual history may allow pre- or intraoperative diagnosis, which is the clue to a less aggressive operation. Postoperative follow up and hormonal therapy are mandatory. PMID- 10793772 TI - [Laparoscopic treatment of varicocele]. AB - The Authors present their experience on the treatment of the varicocele; they performed laparoscopic clamping of the internal spermatic vein in 12 subfertile patients, utilizing only the bipolar coagulation without the use of clips. All patients were available for follow-up, which ranged from 6 to 12 months, including physical examination, Doppler C.W. and study of semen quality. Recurrence of varicocele was not observed, the semen quality demonstrated improvement in semen motility in 9 patients (75%) and normal on 3 patients (25%). The use of bipolar coagulation with smaller ports (phi 5 mm) resulted in less postoperative pain and shorter convalescence than when larger ports were used (phi 10 mm). The Authors conclude that the laparoscopic procedure, used as they do, is safe and costs less than classic ligation with clips as well as radiological occlusion procedures. PMID- 10793773 TI - [The creation of a neovagina with laparoscopic technique]. AB - Vaginal agenesis is sometimes found isolated but more frequently as a part of a malformative syndrome, such as the Rokitansky-Kuster-Hauser or the Morris syndrome. In our department the technique of choice for surgical creation of a neovagina is that described by Vecchietti. A pelviscopic approach for this technique has recently been described. Two cases of vaginal atresia are described; one with Rokitansky-Kuster-Hauser and the other with Morris syndrome, in which the laparoscopic technique was attempted. Due to the aetiopathogenetic and anatomical differences of the two syndromes, the laparoscopic technique resulted easy in the first case but very difficult in the Morris syndrome case, needing laparotomic conversion. We therefore believe that in choosing the surgical approach, a decisive issue is the type of malformation. Indeed in the Morris syndrome, the contiguity between rectum and bladder makes the laparoscopic technique more difficult. PMID- 10793774 TI - [Perihepatic packing combined with wrapping in the treatment of major bi-lobar hepatic trauma]. AB - After the spleen the liver is the most exposed organ to injury in abdominal blunt trauma. The improvement in imaging techniques and in anesthesia and intensive care have contributed to a fallen in mortality rate and an enhanced possibility in non-operative management. In unstable patients the packing may be used to defer the definitive operation. The Authors report a case of liver injury of two lobes where the packing was followed by liver wrapping with an absorbable mesh. The patient, a female aged 67, resulted to have a bilobar liver injury of IV degree in the injury severity score. The delay of reoperation was 15 days. The procedure was complicated by a subphrenic abscess, successfully managed with non operative procedures. Wrapping after liver packing may be considered a good option in unstable patient affected, by severe liver injuries after blunt abdominal traumas. PMID- 10793775 TI - Epidemiological and clinical patterns of diabetes mellitus in Benghazi, Libyan Arab Jamahiriya. AB - The clinical characteristics and prevalence of non-insulin-dependent diabetes mellitus (NIDDM) diagnosed in Benghazi are reported. Data were based on the analysis of records for the period 1981 to 1990. A total of 8922 NIDDM cases (4081 males, 4841 females) were registered during the study period. The overall prevalence rate of NIDDM was 0.19%; it was significantly higher in females (0.21%) than males (0.17%) (P < 0.01). The prevalence rate in patients aged > or = 20 years was 3.8% and was significantly higher in females (4.7%) than in males (2.9%) (P < 0.01). Prevalence rates increased with each higher age group and peaked in the 50-54 years age group. Prevalence of hypertension was 22.5% and prevalence of overweight was 59.4%. The most common complications of diabetes were neuropathy (45.7%), retinopathy (30.5%) and nephropathy (25.2%). PMID- 10793776 TI - Body fat distribution and the risk of non-insulin-dependent diabetes mellitus in the Omani population. AB - Anthropometric measures of overall and central obesity as predictors of non insulin-dependent diabetes mellitus (NIDDM) risk were studied. Data for 4728 Omanis were taken from the 1991 National Diabetes Survey. Diabetes mellitus was assessed using a 2-hour post glucose load. After adjusting for age, sex, family history of diabetes, physical activity and blood pressure, body mass index (BMI) was positively associated with increased risk of diabetes mellitus. Controlling for BMI and other potential confounders, waist-to-hip ratio and waist circumference were positively associated with increased risk of diabetes mellitus. Waist measurement (alone or with hip circumference) is a simple and independent tool for assessing the risk of NIDDM. PMID- 10793777 TI - Psychiatric morbidity in primary care. AB - The prevalence of hidden psychiatric morbidity was assessed using the General Health Questionnaire (GHQ) and Hospital Anxiety Depression Scale (HAD). A total of 149 Bahraini patients aged > or = 16 years were selected randomly from those attending primary health care centres for problems other than psychiatric illness. The prevalence of psychiatric morbidity using GHQ was 45.1% (cut-off > or = 5) and 27.1% (cut-off > or = 9). Using the HAD scale, the prevalence was 44.4% (cut-off > or = 8) and 23.6% (cut-off > or = 11). Psychiatric morbidity was more common in women aged 50-55 years, in divorcees or widows and in lesser educated patients. Either instrument could be used to diagnose psychiatric illness. PMID- 10793778 TI - Psychotropic drugs prescriptions in Al-Qassim region, Saudi Arabia. AB - Psychotropic drugs prescriptions for patients attending psychiatric outpatient clinics were studied. Of the 52,168 prescriptions written in 1996, 18,265 were systematically, randomly selected and evaluated. Incomplete prescriptions were found; the data missing included duration of treatment (18.75%), sex (9.25%), age (8.75%) and diagnosis (7.50%). Antipsychotics (33.1%), antidepressants (23.2%), anticholinergics (22.0%) and anticonvulsants (12.9%) were the most frequently prescribed drugs. Polypharmacy (85%) was the predominant mode of practice. The most common diagnoses were mood (23.1%), anxiety (17.7%) and schizophrenic (16.2%) disorders. Medical education and quality monitoring programmes are suggested to improve the quality of psychotropic prescriptions and modify multiple pharmacotherapy practice. PMID- 10793779 TI - Risk factors of coronary heart disease: attitude and behaviour in family practice in Saudi Arabia. AB - To study risk factors of attitudes and behaviour towards coronary heart disease (CHD), 280 Saudis > or = 20 years attending a family practice answered a structured health and lifestyle questionnaire and had their weight, height, blood pressure and random total cholesterol measured. Significant difference was found between males and females in the mean number of cardiovascular risk factors (t = 3.03, P < 0.01). Few people with high dietary fat intake, obesity or physical inactivity perceived their behaviour as harmful. The number of people who perceived an associated risk to their health increased with incidence of smoking and obesity but not with high fat intake. Physically inactive people were least likely to perceive their behaviour as harmful. PMID- 10793780 TI - Blood pressure patterns among the Omani population. AB - The National Blood Pressure Survey aimed to determine blood pressure levels among Omanis > or = 18 years, study the epidemiological factors most related to blood pressure and suggest a programme for the management and control of hypertension in Oman. Of the 4732 people screened, 1278 (27.01%) had high blood pressure. Multiple regression analysis showed that blood pressure depended on age, body mass index and income per capita simultaneously. In a logistic regression model, adjusted risks associated with obesity and those aged > or = 45 years were significantly increased compared with the relevant different categories. A national plan of action for hypertension control and management is recommended and discussed. PMID- 10793781 TI - Deaths from unintentional injuries in rural areas of the Islamic Republic of Iran. AB - Deaths from accidental injury in the rural areas of 13 provinces in the Islamic Republic of Iran from 1993 to 1994 were investigated. The crude mortality rate was 4.33 per 1000 and the number of deaths from unintentional injuries was 5213 (10.7% of all deaths). There were more deaths among males than females (65.7 per 100,000 versus 26.1 per 100,000). After the age of 1 year, over 65-year-olds had the highest average of deaths resulting from injuries (111.9 per 100,000). The leading causes of death were traffic accidents (55.0%), drowning (10.1%), falls (9.5%) and burns and scalding (9.5%). Since most injuries are preventable, their reduction should be considered a priority. PMID- 10793782 TI - Detection of Mycobacterium tuberculosis complex and non-tuberculous mycobacteria by multiplex polymerase chain reactions. AB - The ability of two-band and three-band multiplex polymerase chain reactions to detect and differentiate Mycobacterium tuberculosis complex from non-tuberculous mycobacteria was evaluated. The polymerase chain reactions differentiated between M. tuberculosis and non-tuberculous mycobacteria when standard strains and clinical isolates of mycobacteria were tested. The sensitivity of the two-band and three-band techniques to detect M. tuberculosis in clinical specimens, compared with smear and/or culture, was 88% and 75% respectively. Although both techniques showed 100% specificity, the superior sensitivity of the two-band technique suggests that it could be more useful in the diagnosis of tuberculosis and in differentiating M. tuberculosis complex from non-tuberculous mycobacteria. PMID- 10793783 TI - Should we routinely check for hepatitis B and C in patients with lichen planus or cutaneous vasculitis? AB - The study aimed to determine the prevalence of HBs-Ag and anti-HCV antibodies in a group of 43 patients with lichen planus and 19 patients with cutaneous vasculitis versus 30 controls. The results showed that 12 (27.9%) patients with lichen planus were positive for HBs-Ag, 9 (20.9%) were positive for anti-HCV antibodies and 3 (7%) were positive for both. In cutaneous vasculitis patients, 3 (15.8%) were HBs-Ag-positive, 7 (36.8%) were anti-HCV-positive and 3 (15.8%) were positive for both. In the control group, 8 (26.7%) were HBs-Ag positive, 3 (10%) were anti-HCV-positive and 1 (3.3%) was positive for both. These values were not statistically significant. PMID- 10793784 TI - Characteristics of visitors to traditional healers in central Sudan. AB - Traditional healing is widespread in Sudan and traditional healers are well respected by the community. This study aimed to assess the characteristics of visitors attending traditional healers, the reasons for visits, the frequency of visits, satisfaction with visits and advantages and disadvantages of visits. The results showed that children under ten years did not take part in visits; most of the visitors were between 21 and 40 years (61%) and were women (62%). Visitors were less educated compared to the general population in the area. The main reasons given for attending traditional healers were treatment (60%) and blessing (26%). Visitors did not mention any disadvantages to visiting traditional healers. PMID- 10793785 TI - [Prevalence of intestinal parasitic disease in three provinces in Morocco]. AB - A study of intestinal parasites was carried out in Taounate, Beni Mellal and Tizinit provinces in Morocco on a sample of 1682 individuals who were representative of the urban and rural area population. For each stool specimen, three microscopic examinations and a Kato test were performed. In the three provinces, about two-thirds of the study participants from rural areas and around 50% of those from urban areas had parasites. The reproducibility of the prevalence figures allowed the study results be generalized to the other provinces of the country. Among the parasite groups encountered, amoebas were the most frequent, followed by flagellates and helminths. Concerning pathogenic species, Entamoeba histolytica, in the amoebic group, was the most commonly found. All the positive cases were treated by specific medication. PMID- 10793786 TI - Infant-feeding practices in Al-Ain, United Arab Emirates. AB - A survey was conducted to study the practices of infant-feeding and the influencing factors in Al-Ain, United Arab Emirates. It involved 375 mothers of different nationalities and backgrounds and 300 healthy infants. The mothers were interviewed at four primary health care clinics in Al-Ain. Results showed that 46% of infants were breastfed for 4-6 months. The mother's nationality and her educational status were significant influences on the mother's decision to exclusively breast-feed the infant, for how long and when to introduce supplementary food. Fresh cow and goat milk were the most common supplements. Inclusion of baby formula as a supplement generally occurred early, perhaps because of advertising and the affluence in Al-Ain. PMID- 10793788 TI - Routine urine analysis in university candidates: is it worthwhile? AB - The study aimed to investigate the abnormalities in urine analysis in university candidates. A total of 247 students (150 males, 97 females) (mean age 20.08 years) had a medical check-up including urine analysis. In all, 38 urine samples had abnormalities (one or more abnormality/urinalysis). Pyuria was found in 20 students (8.1%), haematuria in 15 (6.1%), albuminuria in 12 (4.8%) and casts in 9 (3.6%). No glycosuria was found. Bacteria were found in 22 students (8.9%) and yeasts in two (0.8%). Urinalysis is warranted as a routine test in university students because of the percentage of abnormalities found and its importance in disease surveillance. However, it should not be a criterion for refusal by universities. PMID- 10793787 TI - Setting the scene for an ARI control programme: is it worthwhile in Saudi Arabia? AB - Baseline data on acute respiratory infections (ARI) were collected by a survey questionnaire distributed to physicians of 10% of the health centres randomly selected from each of the five provinces of Saudi Arabia. The physicians estimated that ARI was the cause of sickness in 50% of ill children < 5 years in 1995. None of the physicians had had any training in ARI and they were not aware of any national protocol or programme. Physicians' responses indicated an over use of antibiotics and diagnostic procedures. A national protocol for diagnosis and treatment of ARI has been prepared and distributed and leaders of primary health care and 55 national trainers have been trained. PMID- 10793789 TI - Long-term administration of vitamin A and the process of spermatogenesis. AB - The effect of retinoids on spermatogenesis in adult male gerbils (Gerbillus cheesemani) was studied using light and electron microscopy. Treatment with either 13-cis-retinoic acid or retinol acetate was given for 6 weeks and their effects were compared with controls. It was found that 13-cis-retinoic acid induced almost complete cessation of spermatogenesis and produced alterations in the cytoplasm of Leydig cells. No differences were seen in the testis of animals treated with retinol acetate compared with controls using light microscopy but it appeared to produce noticeable ultrastructural changes in Leydig cells. The changes observed were reversed 12 weeks after stopping treatment. Caution should be exercised regarding the use of dietary retinoids in the prevention of cancer. PMID- 10793790 TI - Isolation of Yersinia enterocolitica from cases of acute appendicitis and ice cream. AB - Seventy (70) appendiceal specimens and 80 ice-cream samples were analysed to detect Yersinia enterocolitica using three different media. Both Y. enterocolitica and Citrobacter freundii were recovered in appendiceal specimens (17.1% and 8.6%) and ice-cream (26.25% and 18.75%) respectively. Thioglycollate medium was more selective and productive in isolating Yersinia. Y. enterocolitica was the major causative agent of acute appendicitis (11/25, 44%). It was sensitive to chloramphenicol, gentamicin, tetracycline and trimethoprim- sulfamethoxazole. PMID- 10793791 TI - The effect of sociodemographic variables on child-spacing in rural Saudi Arabia. AB - Adequate child-spacing is considered a positive factor in the health of mothers and their children. A house-to-house survey of 332 women in Al-Oyaynah village, Saudi Arabia was carried out in April and May of 1995 to determine the existing practice of child-spacing and factors influencing it. The variables examined included age of the mother, age at marriage, education, income, parity, type of infant-feeding and birth order. The age of the mother, age at marriage and education were significantly associated with the length of the birth interval. The current age of mother and her parity were found to be the only significant predictor variables of birth interval. PMID- 10793792 TI - Perspectives on the health care system of the United Arab Emirates. PMID- 10793793 TI - Basic development needs approach in the eastern Mediterranean region. PMID- 10793794 TI - Epidemiological, clinical and haematological profile of schistosomiasis in Yemen. PMID- 10793795 TI - Thrombolytic agents in district hospitals. PMID- 10793797 TI - Mental health services in the Arab world. PMID- 10793796 TI - Case of cryptosporidiosis in an Iraqi woman with ulcerative colitis. PMID- 10793798 TI - Mental health in the Eastern Mediterranean Region of the World Health Organization with a view of the future trends. PMID- 10793799 TI - Relapse among substance-abuse patients in Riyadh, Saudi Arabia. AB - Substance abuse is a recently acknowledged problem in Saudi Arabia. Sociodemographic correlates of substance abuse and relapse rates of a sample of inpatients at Al-Amal Hospital in Riyadh in 1998 were studied. The mean age of patients was 29.5 years with a mean duration of abuse of 9.5 years. Age, unemployment, peer pressure and family and social stresses were factors that showed statistically significant associations with repeat admissions. The most significant predictors of a patient's relapse were unemployment and social stresses. More nation-wide studies are needed to document factors leading to the initiation and continuation of substance abuse and to help treatment and rehabilitative measures. PMID- 10793800 TI - Levels of disability among the elderly in institutionalized and home-based care in Bahrain. AB - We compared the levels of disability between the elderly admitted to an institution and those cared for at home. Of the 74 elderly people in this study, 56 were institutionalized and 18 were living at home. The Clifton Assessment Procedure for the Elderly (CAPE) was used to assess and compare the behavioural disabilities between the two groups. In addition to their younger age, the home cared elderly were less incontinent, more social, better communicators and less confused than the institutionalized group, despite the fact that they had more physical disabilities with regard to bathing and walking. PMID- 10793801 TI - A retrospective audit of electroconvulsive therapy at King Khalid University Hospital, Saudi Arabia. AB - The case records of patients receiving ECT at King Khalid University Hospital over a 10-year period were reviewed for all aspects of ethical and technical administration of ECT. The audit of 127 patients showed some deficiencies in the early years: no seizure monitoring, no training or education for staff (particularly psychiatrists and anaesthetists), and the use of atropine as a premedication for all patients. Recently, regular education and staff training have been organized and seizure timing regularly monitored. Use of an ECT machine with EEG monitoring and the systematic and scaled monitoring of complications is recommended. Results show the necessity of ECT auditing, including an unbiased external audit for effective monitoring of the practical administration of ECT. Similar audits in other mental health institutions in Saudi Arabia are recommended. PMID- 10793802 TI - Motor terminal latency index in carpal tunnel syndrome. AB - We determined the motor terminal latency index (MTLI) of the median nerve across the carpal tunnel in 41 upper extremities of 31 patients with carpal tunnel syndrome. Changes in motor nerve conduction velocity (MNCV), motor terminal latency (MTL), sensory action potential and the amplitude of the compound muscle action potential recorded from the abductor pollicis brevis muscle were all suggestive of proximal and distal segment involvement of the nerve across the carpal tunnel. There was no correlation between forearm MNCV and MTL (r = 0.40), although MTLI was correlated with MTL (r = 0.67) but not with MNCV, indicating a disproportionate conduction across the carpal tunnel. PMID- 10793803 TI - Role of health education programmes within the Libyan community. AB - The effectiveness of existing Libyan health education programmes was evaluated in order to assess how the service may be improved. A representative sample of the general public completed a questionnaire on health knowledge, healthy behaviours and the impact of various health education media. The 872 participants ranked health education media by effectiveness, with television ranked highest and booklets and leaflets lowest. We recommend reorganization of the use of different health education media in future planning. PMID- 10793804 TI - Safer roads for children in the United Arab Emirates. AB - A study was conducted to obtain a representative view of the parents of primary school children regarding which strategies should be employed to reduce the toll of road traffic accidents in the United Arab Emirates. A cross-sectional population-based study was conducted in Al-Ain, UAE, in which parents were asked to give their suggestions on how to make the roads safer for their children. Strategies aimed at reducing vehicle speed, improved personal driving, adherence to existing rules, and driver and pedestrian education were regarded as paramount. There is widespread awareness of road safety problems in the country and the public appears ready to accept major safety initiatives by government agencies. PMID- 10793805 TI - Violence among schoolchildren in Alexandria. AB - Violent behaviour among school students and its predictors were investigated. Selected children (2170) were requested to complete a self-administered questionnaire. Initiating violent assaults in the 18 months prior to the study was reported by 51.0% of boys and 20.9% of girls. Multivariate logistic regression analysis pointed to 16 predictive variables for violent behaviour; few were related to family background whereas the majority were related to the children themselves. Violent assaults were more likely to be initiated by boys and those who were dangerously daring and risk-takers, often fought verbally, threatened to attack others, were cruel to animals, disrupted class discipline, were truant from school or ran away from home and were disciplined by corporal punishment by their parents and their teachers. School-based prevention and intervention programmes addressing modifiable predictors should be considered. PMID- 10793806 TI - Nucleolus organizer region heteromorphism in patients with Down syndrome and their parents. AB - The study aimed to evaluate the role of nucleolus organizer region (NOR) heteromorphism as an etiological factor for parental nondisjunction in Down syndrome by comparing 25 patients affected by Down syndrome, and their parents with a control group of 80 non-affected Egyptians. All parents had normal karyotypes. The average modal number per parent of Ag-positive NORs was significantly higher in parents than controls. A significant difference in the size of the double-NOR variants (dNORs) was found. The mean maternal and paternal ages were significantly lower, with a significant increase in spontaneous abortions, for dNOR(+) couples compared with dNOR(-) couples. PMID- 10793807 TI - Correlates of age at natural menopause: a community-based study in Alexandria. AB - Differences in the age at natural menopause were examined using a retrospective population sample of 289 naturally menopausal women. The mean age at natural menopause was 46.70 +/- 5.44 years. Earlier menopause occurred in women living in semiurban areas, divorced/separated and less educated women, and women who were younger at: first marriage, widowhood, divorce/separation and first or last full term pregnancy. Later menopause occurred in women who had: irregular menstrual periods before 25 years, dysmenorrhoea and mid-cycle spotting. Duration of oral contraceptives use, weight and body mass index were significantly positively correlated with age at natural menopause. Multiple regression analyses indicated that age at last full-term pregnancy, residence, pattern of menstrual cessation and duration of oral contraceptive use were the significant predictors of the end of menstrual activity. PMID- 10793808 TI - The use of medication in infants in Alexandria, Egypt. AB - In Egypt, the determinants and rationale for drug use among infants have not been previously studied. We give an overview of the use of prescribed and non prescribed medication over a 1-month period in infants in Alexandria, Egypt. In 89.6% of cases mothers stated that their infants had suffered some ailments which necessitated intervention; 27.5% used traditional therapies or non-prescribed medication and 72.5% sought medical advice. The non-prescribed medicines used were mainly antispasmodics and antipyretics. This study provides the first overview of the frequency of use of non-prescribed medication in infants and detected some indicators of irrational drug prescribing by physicians. PMID- 10793809 TI - [Antibiotic therapy in general medicine in Monastir, Tunisia]. AB - In order to determine the cost and frequency of antibiotic prescription by general practitioners, we studied 563 outpatients from health centres in Monastir (Tunisia). All patients had acute diseases. Antibiotics were prescribed to 50.4%. Single antibiotics were generally prescribed, but 52.8% of these patients did not have any laboratory tests. The more frequently used antibiotics were penicillin G and A. Antibiotics cost represented 34.7% of medicinal cost borne by patients and 49.7% of the cost borne by the public sector. Rationalization of medicinal prescription would have a positive impact on household and state budgets. PMID- 10793810 TI - Fungal infection of burn wounds in patients with open and occlusive treatment methods. AB - Fungal infection of burn wounds was investigated in a prospective study of 130 patients managed either with open or occlusive treatment methods. In all, 30 fungal isolates were recovered from 26 patients all of whom had bacterial infection also, except for one patient. The predominant fungi recovered were Aspergillus spp. and Candida spp. Fungal infection was more common in patients treated with open dressing (25.5%) than occlusive dressing (16.0%). Fungal culture from tissue specimens gave a better isolation rate of fungi than from cotton swab specimens. PMID- 10793811 TI - Prevalence of malocclusion in a sample of Lebanese schoolchildren: an epidemiological study. AB - The study aimed to provide oral health planners in Lebanon with information on the prevalence of malocclusion in schoolchildren in an attempt to define the target population for orthodontic services in the future. A total of 851 schoolchildren (446 males and 405 females) aged 9-15 years were examined for malocclusion using Angle's classification. In all, 59.5% of the sample had malocclusions, 35.5% of which were of dental origin and 24% had skeletal discrepancy (19% Class II and 5% Class III malocclusions). A statistically significant difference was found between males and females. The findings are compared with those of studies of other ethnic groups and suggestions for future epidemiological research are presented. PMID- 10793812 TI - Iron status of Libyan infants with urinary tract infection. AB - Serum iron, serum total iron binding capacity and tranferrin saturation levels were measured in 45 infants, of whom 19 had urinary tract infections confirmed by positive suprapubic aspiration. The control group comprised 26 healthy infants with negative results. Mean serum iron and transferrin saturation values were significantly lower in infants with urinary tract infection compared with the control group (P < 0.01 and P < 0.001 respectively), especially if the cultured organism was Escherichia coli (P < 0.01, P < 0.02). Total iron binding capacity was within the normal range. We conclude that iron deficiency is not a major factor in the etiopathogenesis of urinary tract infection in Libyan infants. PMID- 10793813 TI - Bronchoconstrictor effect of exercise in healthy Libyan children in Tripoli. AB - To study the effects of short-term exercise on pulse rate and peak expiratory flow rate in healthy Libyan schoolchildren, 650 healthy students (330 boys, 320 girls) aged 4.5 years to 14.9 years were selected from four randomly chosen Tripoli primary schools. Pulse rate and peak expiratory flow rate were measured at rest in standing position and immediately after rhythmic short-term exercise. Exercise markedly increased pulse rate (mean differences being significantly higher in girls than boys) and markedly reduced peak expiratory flow rate (mean difference being significantly higher in boys than girls). We found 10% of the children had a reduction in peak expiratory flow rate > or = 15% from the baseline. PMID- 10793814 TI - Eating disorders: a transcultural perspective. PMID- 10793815 TI - Mental health manpower development in Afghanistan: a report on a training course for primary health care physicians. PMID- 10793816 TI - Integration of mental health into primary care in Al-Qassim Region, Saudi Arabia: planning phase I. PMID- 10793817 TI - Integration of mental health into primary care in Al-Qassim Region, Saudi Arabia: curriculum development II. PMID- 10793818 TI - Secnidazole response in amoebiasis and giardiasis. PMID- 10793819 TI - Vaccine trial against canine visceral Leismaniasis in the Islamic Republic of Iran. PMID- 10793820 TI - The newly defined burden of mental problems. PMID- 10793821 TI - Breast cancer: patient characteristics and survival analysis at Salmaniya medical complex, Bahrain. AB - Breast cancer is the most common malignant neoplasm affecting women in Bahrain. We studied the demographic characteristics and clinical presentation of patients with breast cancer and conducted survival analyses based on various factors. Data on all the 117 breast cancer patients at the Salmaniya Medical Complex who had been discharged during the period 1982-94 were audited. When first seen by a physician 70% of the patients had a lump size > 2 cm, with 51.3% presenting in clinical stage II, 21.4% in clinical stage III and 11.1% in clinical stage IV. Only 6.8% of patients presented in clinical stage I. Cumulative survival rates calculated using the Kaplan-Meier method were 68.8%, 57.33% and 36.44% after 5, 7 and 10 years respectively. Our study highlights the need for increased awareness about breast cancer by women in Bahrain, and the need for early detection through regular screening. PMID- 10793822 TI - First cigarette smoking experience among secondary-school students in Aden, Republic of Yemen. AB - The objectives of this study were first to estimate the prevalence of cigarette smoking among secondary-school students and then to identify and analyse the demographic, social and cultural risk factors associated with the first cigarette smoking experience. A sample comprising 1000 students was selected randomly. Results showed that 19.6% of the total sample smoked; 15.5% among females and 21.9% among males. Family and/or friends appeared to influence the first cigarette experience and thus prevention strategies should involve not only the students themselves but the home, school and social environments also. PMID- 10793823 TI - Maternal smoking and breastfeeding. AB - A 2-month study was undertaken to investigate the effects of maternal smoking on breastfeeding. A total of 500 mothers were interviewed twice postpartum and information about maternal smoking and sociodemographic factors was collected. After adjustment for maternal smoking and other confounders, results suggested that smoking had a direct effect on breastfeeding. The prevalence of breastfeeding reduced significantly among smokers but there was no significant reduction among non-smokers. Education and social class were related positively to continued breastfeeding but negatively to smoking. Because breastfeeding decreased with smoking, smoking should be discouraged, particularly in developing countries where breastfeeding constitutes an essential child survival measure. PMID- 10793824 TI - Haemolytic potential of three chemotherapeutic agents and aspirin in glucose-6 phosphate dehydrogenase deficiency. AB - The potential haemolytic effect of three chemotherapeutic drugs and aspirin was tested in vitro by gluthathione stability tests. Blood was collected from the local population of Basra, Iraq where previous studies had found a high frequency of glucose-6-phosphate dehydrogenase (G6PD) deficiency. Primaquine, chloramphenicol and sulfanilamide caused significant concentration-dependent reductions of glutathione levels in G6PD-deficient red cells when compared to normal red cells. Acetylsalicylic acid had no effect on glutathione level. The G6PD-deficient erythrocytes behaved as previously reported, probably due to similar patterns in the distribution of its variants. Studies on each local variant are warranted and new drugs should be tested for haemolytic potential prior to their introduction in areas where the deficiency is common. PMID- 10793826 TI - A diagnostic clinical genetic study of craniofacial dysmorphism. AB - A diagnostic evaluation of craniofacial anomalies, either isolated or as part of a genetic syndrome was conducted on 25 patients (8 females, 17 males), age range 2 months to 47 years. Complete genetic examination, pedigree analysis, anthropometric measurements and radiological studies were carried out. Cytogenetic studies included fluorescence in situ hybridization (FISH) when indicated. In all, 15 patients had chromosomal abnormalities. Five patients had unbalanced chromosome rearrangements and six had chromosome markers. Three patients were FISH-positive for William syndrome and one was positive for Prader Willi syndrome. Ten patients had monogenic disorders. Five were diagnosed as craniosynostosis syndromes. We conclude that minor features are useful for making a diagnosis of congenital anomalies. PMID- 10793825 TI - Acute graft-versus-host disease in thalassaemic marrow transplantation with low dose antithymocyte globulin. AB - Our unit performed transplantations on 21 classes II and III thalassaemic patients (class II patients had either hepatomegaly or portal fibrosis and class III patients had both). We used busulfan (15 mg/kg) and cyclophosphamide (200 mg/kg). Graft-versus-host disease (GVHD) prophylaxis was cyclosporin, prednisolone and low-dose antithymocyte globulin. Our patient data showed a low incidence of acute GVHD following transplantation. We offer this regimen as an acceptable therapy for thalassaemic patients undergoing allogeneic marrow transplantation as a safe clinical procedure, irrespective of the class of patient. PMID- 10793828 TI - CK13 in craniopharyngioma versus related odontogenic neoplasms and human enamel organ. AB - The monoclonal antibody NCL-CK13 was studied in specimens of craniopharyngioma, ameloblastoma and calcifying odontogenic cyst neoplasms and the mandible and maxillae of normal human fetuses. There was a decrease in NCL-CK13 as the dental lamina developed, with a complete loss in the enamel organ. The neoplastic epithelia of the neoplasms revealed a clear phenotypic and immunohistochemical reactive relationship to the stratified embroyonic mucosa, away from the enamel organ. This suggests that these neoplasms might have their histogenesis from early stage epithelium, the oral part of the dental lamina or its remnants. PMID- 10793827 TI - Immunohistochemical detection of p53 protein in ameloblastoma types. AB - Overexpression of p53 protein in unicystic ameloblastoma (uAB) is denser than in the conventional ameloblastoma (cAB) type, indicating increased wild type p53- suppressing the growth potential of uAB and denoting the early event of neoplastic transformation, probably of a previous odontogenic cyst. Overexpression of p53 in borderline cAB and malignant ameloblastoma (mAB) types might reflect a mutational p53 protein playing an oncogenic role, promoting tumour growth. Overexpression of p53 protein could be a valid screening method for predicting underlying malignant genetic changes in AB types, through increased frequency of immunoreactive cells or increased staining density. PMID- 10793829 TI - The translation into Arabic and revalidation of a fatigue questionnaire. AB - The aim of this study was to translate a fatigue questionnaire, which had been developed in England for use in epidemiological studies and in community settings, into Arabic. It was intended that the translated questionnaire could be used in any setting where Arabic is the first language of the patient. The process of translating the questionnaire and the revalidation method are described. The Arabic translation was shown to be both reliable and valid in the United Arab Emirates setting. PMID- 10793830 TI - Rethinking the health services insurance system: a new model for Iranian railroad households. AB - During 1995 and 1996, a 10% stratified random sample of 2124 people representing permanent Iranian railroad workers in 12 provinces was selected to survey the use of health services by personnel and their families. Data were collected from 2107 workers through interviews using a structured questionnaire. The objective was to determine demographic and health characteristics and user satisfaction in order to improve the present delivery of the health insurance system. A new plan for the delivery of health insurance, which focuses upon social accountability and community orientation and is based on the roles of family doctors and nurses in railroad health posts, is proposed and evaluated. PMID- 10793831 TI - Probability of contraceptive continuation and its determinants. AB - Probability of contraceptive continuation and factors associated with discontinuation were assessed in a retrospective study of women's records in Alexandria. Contraceptive continuation rates were 88/100, 75/100 and 56/100 women at 6, 12 and 24 months respectively. Reasons for discontinuation included side effects and health concerns (30.4%), accidental pregnancy and personal reasons (6.4%) and desire for pregnancy (5.4%). Women who were older and who had several children were most likely to continue contraceptive use. Three significant predictors of discontinuation were side-effects and health concerns, age of the woman and contraceptive method. Women should be fully informed of the contraceptive methods available and their side-effects, and counselling programmes should be offered, particularly during the first 2 years of use. PMID- 10793832 TI - Outbreak of pertussis in Basra, Iraq. AB - A total of 133 pertussis cases were studied during an outbreak in Basra from June to December 1996. Most were females and were immunized. Bordetella spp. was isolated in 48.1% of the cases. The isolation rate was highest among infants and decreased with increasing age, and was highest during the catarrhal stage. B. pertussis was the most common species; however, B. parapertussis infection did occur. There were some severe cases of pertussis among infants caused mainly by B. pertussis and dual Bordetella infection. Infection was transmitted by close contact with a pertussis case. PMID- 10793833 TI - Assessment of periodontal disease using the CPITN index in a rural population in Ninevah, Iraq. AB - To estimate the prevalence of periodontal disease in a rural population and to assess the periodontal treatment needs, a sample of 1418 individuals aged between 7 years and 70 years, from three villages in Ninevah Governorate was examined using the community periodental index of treatment needs to assess their periodontal condition. Only 12.5% of the sample had healthy teeth and gums, and calculus was the most frequently observed periodontal condition. There is a need for dental prophylaxis and instruction in the use of oral hygiene procedures. There was no significant difference between males and females in periodontal health. PMID- 10793834 TI - Sinusitis as a cause of orbital cellulitis. AB - To determine the importance of sinusitis as a cause of orbital cellulitis, the causative organisms and peak age of occurrence, 25 patients hospitalized with orbital cellulitis (ages 8 months to 17 years; 80%, 1-4 years) were studied. Complete blood counts were carried out and radiographic sinus examinations and eye swabs (for culturing) performed prior to antibiotic treatment. Sinusitis was evident in 72% of the patients. Eye swab cultures indicated 80% had streptococcal, staphylococcal or enterococcal infection. The data indicate that sinusitis is an important cause of orbital cellulitis. The most common causative organisms were Streptococcus viridans (44%) and Staphylococcus aureus (32%). Initial antibiotics should therefore cover both organisms. PMID- 10793835 TI - Peak expiratory flow rate nomogram in Libyan schoolchildren. AB - The study aimed to develop a peak expiratory flow rate nomogram for Libyan children. Of 900 children randomly selected from four Tripoli primary schools, 670 (330 girls and 340 boys) with age range 4.5-14.9 years, fulfilled the selection criteria. Peak expiratory flow rate (PEFR) was recorded in a standing position using a mini-Wright peak flow meter. Anthropometric measurements, weight, height, head circumference and mid-upper-arm circumference were recorded and surface area and body mass index were calculated. Our findings showed PEFR to be significantly related to height (r = 0.74), age (r = 0.70), surface area (r = 0.64) and weight (r = 0.62): P < 0.001. The PEFR nomogram in Libyan children differed from the British standard, which highlights the need for a local reference nomogram. PMID- 10793837 TI - Working with families to reduce the risk of home accidents in children. PMID- 10793836 TI - [Value of vaccination data analysis at the district level]. AB - This study aimed to demonstrate the importance of analysing local immunization data to improve performance of national prevention programmes. From the immunization registers kept in basic health centres, we identified 1271 children receiving their first vaccine at 3 months. Examination of the age at the subsequent vaccinations and time intervals between vaccinations showed that 81.9% had received all the vaccines required by the Ministry of Health but only 48.5% had received immunization meeting the requirements for age and time interval. The analysis also helped identify health centres with best performance (fewer children lost to follow-up and better implementation of immunization schedule) and those vaccinations needing the greatest attention from health professionals. PMID- 10793838 TI - Behavioural interventions as aids for asthmatic patients: a review. PMID- 10793839 TI - Twenty years of family medicine education in Saudi Arabia. PMID- 10793840 TI - Review of the history of the teaching of medicine in Arabic. PMID- 10793842 TI - Koro-like syndrome in a Jordanian male. PMID- 10793841 TI - Selected haematological and lipid profiles of Libyan children up to two years of age in Benghazi. PMID- 10793843 TI - Strengthening rural health services project. PMID- 10793844 TI - Plasticizers in total diet samples, baby food and infant formulae. AB - The plasticizers di-n-butylphthalate (DBP), butylbenzylphthalate (BBP), di-2 (ethylhexyl)phthalate (DEHP) and di-2-(ethylhexyl)adipate (DEHA) were analysed in 29 total diet samples, in 11 samples of baby food and in 11 samples of infant formulae. In all of the total diet samples the presence of one or more of the plasticizers was demonstrated. Maximum and minimum mean concentrations in the total diet samples were: 0.09-0.19 mg DBP/kg, 0.017-0.019 mg BBP/kg, 0.11-0.18 mg DEHP/kg and 0.13-0.14 mg DEHA/kg. One or more of the phthalates was also found in about 50% of the samples of baby food as well as in infant formulae. The calculated mean maximum intakes of the individual compounds from the total diet samples were below 10% of the restrictions proposed by the EU Scientific Committee for Food (SCF), and the spread in individual intakes was considerable. DEHP was the plasticizer determined most frequently and contributed the highest fraction of its tolerable daily intake (TDI). Hence, the maximum calculated intake of DEHP from single samples of the foodstuffs analysed could be up to one third of the TDI. The calculated mean intake of DEHA was about 1% of the TDI with a maximum value of 13% of the TDI. Violations of the restrictions proposed by the EU Scientific Committee for Food (SCF) in the form of TDI values or specific migration limits were not found in this investigation. PMID- 10793845 TI - Br concentration as an indication of pre-baking bromation of bread products. AB - Br concentration in bread for baked bread products was shown to be linearly proportional to the amount of Br added per kg of flour used to make the product. Br concentration in bread can be used to help identify those bread products with the greatest likelihood of containing bromate residues. Instrumental neutron activation analysis was used to determine Br in test portions of bread products from commercial bakeries, homemade bread, flour, and unbaked dough. High performance liquid chromatography was used to determine the bromate residue in selected test portions. PMID- 10793846 TI - Beryllium in food and drinking water--a summary of available knowledge. AB - In an overall evaluation of the situation published by IARC in 1993, beryllium and beryllium compounds are identified as carcinogens to humans. This prompted the initiation of this study on beryllium which reviews the situation up to 1998 on the aspects: properties and applications, toxicity, analytical procedures for food and drinking water, reference materials, occurrence in food and drinking water and estimates of daily dietary exposure. Special emphasis is put on analytical aspects and levels of beryllium in food and drinking water. PMID- 10793847 TI - Liquid chromatographic method for fumonisin B1 in sorghum syrup and corn-based breakfast cereals. AB - The fungus Fusarium verticillioides has been found on corn and sorghum, so it is possible that one or more of these toxins may be found in corn products such as breakfast cereals and syrup prepared from sorghum. Published methods when applied to syrups spiked with fumonisins gave low recoveries, less than 50%. A method was therefore developed which would be applicable to the syrup and breakfast cereals as well. Test samples were extracted with methanol-0.1 M sodium phosphate buffer (pH3) (1 + 1). The extract was diluted with water and applied to a 1 g C18 column. The column was washed with acetonitrile-water (2 + 8). Fumonisin B1 (FB1) was eluted with acetonitrile-trifluoroacetic acid (1000 + 1). The purified extract was evaporated and the toxin was derivatized with ophthaldialdehyde mercaptoethanol. The reaction mixture was resolved on a C18 liquid chromatographic column using acetonitrile-water-acetic acid (500 + 550 + 10.5) as the mobile phase at 37 degrees C, and FB1 measured with a fluorescence detector (excitation 335 nm, emission 440 nm). Recoveries of FB1 added to samples of sorghum syrup at levels ranging from 0.1 to 1.0 microgram/g were 94-132%. Recoveries of FB1 added to samples of breakfast cereal (corn flakes) at levels ranging from 0.2 to 1.6 micrograms/g were 96-100%. The method was applied to the analysis of 35 samples of sorghum syrup collected from 15 states in the US. One sample was found to contain FB1 at 0.12 microgram/g. A total of 32 samples of breakfast cereals collected by the Food and Drug Administration inspectors from grocery stores around the Kansas City area were analysed; no FB1 was found in the breakfast cereals (< 0.01 microgram/g). Results of this study indicated that FB1 possibly is not a problem in sorghum syrup and corn-based breakfast cereals in the US. PMID- 10793848 TI - Transfer of ochratoxin A during lactation: exposure of suckling via the milk of rabbit does fed a naturally-contaminated feed. AB - The transfer of ochratoxin A from the blood to the milk of lactating rabbit does and subsequently the exposure of their sucklings to the mycotoxin were investigated. An effective transfer of ochratoxin A from blood to milk was shown in lactating rabbit does fed a naturally-contaminated diet (10-20 g/kg of body weight/day) throughout a lactation period of 19 days. The ochratoxin A concentrations in plasma and in milk did not significantly change throughout the lactation period with a mean milk/plasma concentration ratio of 0.015. These variables were however significantly correlated (p < 0.05), as were the ingested amounts and milk concentrations. At slaughter, the highest concentration of ochratoxin A accumulated in the body of the rabbit does were found in kidney (1.2 g/kg) followed by liver (158 ng/kg), mammary gland (105 ng/kg) and muscle (38 ng/kg). A linear relationship was found between the ochratoxin A concentrations in milk and in the plasma of the sucklings, indicating an effective transfer of the toxin to the sucklings. If the same is true in humans, the exposure of the breast-fed infant to the toxin, which has been largely reported in the literature, should be a major matter of concern for human health. PMID- 10793849 TI - Food Additives and Contaminants started in 1984. PMID- 10793850 TI - Detection of adulteration of locust bean gum with guar gum by capillary electrophoresis and polarized light microscopy. AB - Capillary electrophoresis (CE) and polarized light microscopy (PLM) were utilized in the detection of the adulteration of locust bean gum with guar gum. For CE analyses, standards of locust bean and guar gums were extracted with 30% CH3CN, removing the residual proteins from the gum matrix. A 8.75 mM NaH2PO4-20.6 mM Na2B4O7 buffer, pH 9, was used to separate these proteins and to identify marker proteins that were present in the guar gum. These markers did not co-migrate with components in the extracts of mechanically processed locust bean gum, and are used as indicators of adulteration. Using PLM with toluidine blue and iodine staining techniques, unadulterated locust bean gum samples were distinguished from mixed samples through the differential staining of components in locust bean versus guar and tara gums. These experiments in the use of CE and PLM provide orthogonal and complementary methods for the verification of 'true' positives and the elimination of 'false' positives. PMID- 10793851 TI - Shelf-life extension of cod fillets with an acetate buffer spray prior to packaging under modified atmospheres. AB - Fresh cod fillets (Gadus morhua) were sprayed with a 10% acetate buffer (pH 5.6), packed with an industrial gas-flushing packaging machine under modified atmospheres (50% CO2--45% O2--5% N2, 2 cm3/1 g gas/product ratio) and stored at 7 degrees C for 12 days. Control cod fillets were directly packed and stored under the same conditions. A reduction of the aerobic plate counts was observed immediately after the cod fillets had been sprayed. During storage under modified atmospheres, there was complete inhibition of H2S-producing bacteria and Enterobacteriaceae in the treated cod fillets. Production of total volatile bases and trimethylamine (TMA) was inhibited in treated fillets for 10 days' storage under modified atmospheres. Inhibition of TMA production can be attributed to growth inhibition of H2S-producing bacteria, inhibition of the trimethylamine oxide (TMAO)-dependent metabolism of TMAO-reducing bacteria and the stable pH during storage. The shelf-life, at 7 degrees C, of treated cod fillets, based on cooked flavour score, was almost 12 days, ca 8 days more than shelf-life of the control fillets. PMID- 10793852 TI - Study of several aspects of a general method for the determination of polycyclic aromatic hydrocarbons in liquid smoke flavourings by gas chromatography-mass spectrometry. AB - The effectiveness of the steps of a general method, which includes alkaline treatment of the samples, extraction, clean-up and analysis, for the determination of polycyclic aromatic hydrocarbons in liquid smoke flavourings and smoked foods have been studied, by using mixtures of pure polycyclic aromatic hydrocarbons and of smoke flavouring compounds. All these products were applied to samples of liquid smoke flavourings in order to test the effectiveness of each step of the method, as well as to investigate its effect on real samples, and thus obtain a suitable procedure for the study of polycyclic aromatic hydrocarbons in liquid smoke flavourings. The alkaline treatment has been proved to be necessary because of its capability for removing some smoke components. Both cyclohexane and dichloromethane are able to extract polycyclic aromatic hydrocarbons with high recovery percentages; however, cyclohexane is more adequate due to its lower ability to extract other smoke flavourings components. It is important to note that the elution sequence of polycyclic aromatic hydrocarbons during the clean-up step is quite different depending on the solvent used to dissolve the extract, which determines the ability of this process to isolate polycyclic aromatic hydrocarbons for their accurate identification and quantification by gas chromatography-mass spectrometry in selected ion monitoring mode. Finally, the PAHs present in a liquid smoke flavouring and their concentrations were determined; it was observed that, despite the different extraction solvents and clean-up procedures used, the results were very similar. PMID- 10793853 TI - Trace metals in soft and durum wheat from Italy. AB - A survey was carried out with the aim to assess the levels of some toxic (cadmium, lead) and essential (copper, zinc) trace metals in wheat grown in Italy. A total of 178 samples of soft wheat grain and 239 samples of durum wheat grain from all the Italian wheat-growing regions were pooled into 35 and 38 representative samples respectively. After dry ashing, cadmium and lead were determined by graphite furnace atomic absorption spectrometry (GFAAS), whereas copper and zinc were determined by flame atomic absorption spectrometry (FAAS). In soft wheat the mean and median contents of all samples were (on a dry weight basis) 40 and 33 micrograms kg-1 for cadmium, 16 and 14 micrograms kg-1 for lead, 3.4 and 3.2 mg kg-1 for copper, 33 and 32 mg kg-1 for zinc. Similar levels were found in durum wheat. In this latter case the mean and median were 42 and 39 micrograms kg-1 for cadmium, 15 and 14 micrograms kg-1 for lead, 3.5 and 3.2 mg kg-1 for copper, 34 and 34 mg kg-1 for zinc. Significant differences were detected for some metals in relation to geographical provenance and variety. The average intake of the four selected elements from wheat-based products was estimated for the Italian population. PMID- 10793854 TI - Fumonisin levels in commercial corn products in Buenos Aires, Argentina. AB - Fumonisins B1 (FB1) and B2 (FB2) were determined in 35 samples of corn flour and corn grits destined for human consumption and purchased directly from Buenos Aires food shops and supermarkets from October 1996 to January 1997 and during the month of January 1998. During the first period of sample collecting, 16 out of 19 samples were found to be contaminated. Considering all 19 samples, contamination levels were between not detected and 1860 ng/g FB1, and from not detected to 768 ng/g FB2. During the second period all 16 samples were found to be contaminated with levels ranging from 75 to 4987 ng/g FB1, and from not detected to 1818 ng/g FB2. The levels of FB1 and FB2 in the samples collected during January 1998 were significantly higher than the samples collected during the period from October 1996 to January 1997. No significant difference was found in terms of fumonisin levels between the branded and unbranded samples. PMID- 10793855 TI - Occurrence of aflatoxin M1 in Korean dairy products determined by ELISA and HPLC. AB - The occurrence of aflatoxin M1 (AFM1) in pasteurized milk and dairy products was investigated by using direct competitive enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC). The recoveries of AFM1 from the samples spiked at levels between 5 and 500 pg/ml were 88.0-106.5% for pasteurized milk and 84.0-94.0% for yoghurt by ELISA. By HPLC, the recoveries were 103-120% for pasteurized milk and 87.0-93.0% for yoghurt. The limits of detection were found to be 2 pg/ml by ELISA and 10 pg/ml by HPLC. Among a total of 180 samples collected in Seoul, Korea, the incidence of AFM1 in pasteurized milk, infant formula, powdered milk and yoghurt was 76, 85, 75, and 83%, respectively, with a mean concentration of 18, 46, 200, and 29 pg/g, respectively, when determined by ELISA. These results obtained by ELISA were closely related to those by HPLC for AFM1 (r2 = 0.9783). PMID- 10793856 TI - Evaluation and validation of two fluorometric HPLC methods for the determination of aflatoxin B1 in olive oil. AB - Two methods for the determination of aflatoxin B1 (AFB1) in olive oil were tested and compared. In method A the oil sample was mixed with methanol + water (60 + 40), extracted with hexane and then with chloroform. Chloroform was evaporated and the residue was dissolved with dichloromethane which was then transferred for clean-up onto a silica 'Sep-Pak' cartridge. The cartridge was pre-washed with hexane, ethyl ether and dichloromethane. AFB1 was eluted with chloroform + acetone (9 + 1) and evaporated to dryness. In method B, the oil sample was mixed with methanol + water (80 + 20), shaken and centrifuged. The supernatant was diluted 1:10 with water and 10 ml of the diluted mixture transferred to an 'Aflaprep' immunoaffinity column for the clean-up step. AFB1 was eluted with acetonitrile and evaporated to dryness. AFB1 from both methods was derivatized to its hemiacetal (AFB2a) and then quantitated by HPLC using a C18 (60 A 4.6 x 250 mm) column with fluorescence detection. Both methods are simple, reliable and efficient, but method A showed a lower detection limit (2.8 ng/kg) than method B (56 ng/kg). With a 95% confidence level there was no significant difference in recovery between the two methods, which was 87.2% for method A and 84.8% for method B. In addition, application of a two-tailed F-test to the variances within spiked samples at concentrations 1, 2, 5 and 10 micrograms/kg separately showed that there was no significant difference in the precisions of the two methods. Fifty samples of olive oil of Greek origin produced between 1995 and 1998 were examined with both methods for the presence of AFB1. When analysing the samples with method B, the presence of AFB1 was not detected. The use of method A revealed the presence of AFB1 in 72% of the samples. The range of contamination was generally found to be very low (2.8-15.7 ng/kg), however one sample was contaminated with 46.3 ng/kg. PMID- 10793858 TI - European priorities for research to support legislation in the area of food contact materials and articles. AB - A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years. PMID- 10793857 TI - Furazolidone residues in pigs: criteria to distinguish between treatment and contamination. AB - The use of furazolidone in food-producing animals has been banned in the EU. The ban can most effectively be enforced by monitoring for bound residues containing the 3-amino-2-oxazolidinone (AOZ) moiety. Unlike the parent drug, AOZ residues are stable and can be detected for prolonged periods after cessation of treatment. However, AOZ can be passed from pig-to-pig following brief exposure of unmedicated animals to housing that previously contained medicated pigs. We describe criteria by which a distinction may be drawn between pigs treated illegally with the drug and pigs that contain detectable AOZ residues as a result of exposure to contaminated housing. These criteria are that illegally treated pigs will have a concentration ratio of AOZ in bile:kidney of less than 0.3; while unmedicated pigs will have a concentration ratio of AOZ in bile:kidney of greater than 3.0. Using this criteria, 12 pigs, either treated with the drug or exposed to contaminated housing were analysed in a blind study. The pigs were classified as 'Treated' or 'Contaminated' on the basis of the criteria described above. All 12 pigs were assigned to the correct group. This shows that it is possible to differentiate between furazolidone abuse and contamination. PMID- 10793859 TI - Multiple sclerosis and the eye. PMID- 10793860 TI - Human intraocular pressure and its diurnal variation in healthy subjects. PMID- 10793861 TI - The optometric assessment of the visually impaired infant and young child. PMID- 10793862 TI - Clinical assessment of anterior chamber depth. PMID- 10793863 TI - The immune system and the eye. PMID- 10793864 TI - Silicone oil negated the need for an aphakia contact lens. PMID- 10793865 TI - Assessment of patients with age-related cataract. PMID- 10793866 TI - Launch of the bone and joint decade 2000-2010. PMID- 10793867 TI - Effects of alendronate on bone density in men with primary and secondary osteoporosis. AB - Alendronate has been reported to increase bone mineral density (BMD) and reduce fracture risk in women with osteoporosis. As there are no proven safe and effective treatments available for men with osteoporosis, we compared the effects of alendronate (10 mg/day) on BMD, measured using dual-energy X-ray absorptiometry, in a 12-month prospective, controlled, open label study involving (i) men with primary (n = 23) or secondary osteoporosis (n = 18), (ii) postmenopausal women with primary (n = 18) or secondary (n = 21) osteoporosis, and (iii) 29 male and 14 female untreated controls matched by age, height and weight. The patients had one or more vertebral fractures and ranged in age from 34.6 to 85.1 years. BMD was detectably increased relative to baseline by 6 months, and increased by comparable amounts in males and females with primary or secondary osteoporosis. At 12 months, lumbar spine BMD was 5.4% +/- 1.1% to 7.0% +/- 2.2% higher in the treated groups compared with baseline and controls (p < 0.05 to 0.0001). Trochanteric BMD increased by 2.6% +/- 1.5% and 3.7% +/- 1.7% in treated men with primary and secondary osteoporosis, respectively (p = 0.06 to 0.08), and by 3.9% +/- 1.3% in treated women with primary osteoporosis (p < 0.01) after 12 months. No significant changes were detected at the femoral neck or Ward's triangle. BMD remained unchanged in controls. We infer that alendronate has comparable incremental effects on BMD in men and women with primary and secondary osteoporosis within 12 months of treatment. The changes are in the order of 0.5 SD--effects associated with a clinically worthwhile reduction in fracture risk. The data provide room for optimism regarding the role of alendronate in the treatment of osteoporosis in men. Randomized, double-masked and placebo-controlled trials are needed to confirm these preliminary findings and demonstrate antifracture efficacy using vertebral and nonvertebral fracture rates as the primary endpoint. PMID- 10793868 TI - Sex difference in the validity of vertebral deformities as an index of prevalent vertebral osteoporotic fractures: a population survey of older men and women. AB - Morphometric methods have been developed for standardized assessment of vertebral deformities in clinical and epidemiologic studies of spinal osteoporosis. However, vertebral deformity may be caused by a variety of other conditions. To examine the validity of morphometrically assessed vertebral deformities as an index of osteoporotic vertebral fractures, we developed an algorithm for radiological differential classification (RDC) based on a combination of quantitative and qualitative assessment of lateral spinal radiographs. Radiographs were obtained in a population of 50- to 80-year-old German women (n = 283) and men (n = 297) surveyed in the context of the European Vertebral Osteoporosis Study (EVOS). Morphometric methods (Eastell 3 SD and 4 SD criteria, McCloskey) were validated against RDC and against bone mineral density (BMD) at the femur and the lumbar spine. According to RDC 36 persons (6.2%) had at least one osteoporotic vertebral fracture; among 516 (88.9%) nonosteoporotics 154 had severe spondylosis, 132 had other spinal disease and 219 had normal findings; 14 persons (2.4%) could not be unequivocally classified. The prevalence of morphometrically assessed vertebral deformities ranged from 7.3% to 19.2% in women and from 3.5% to 16.6% in men, depending on the stringency of the morphometric criteria. The agreement between RDC and morphometric methods was poor. In men, 62-86% of cases with vertebral deformities were classified as nonosteoporotic (severe spondylosis or other spinal disease) by RDC, compared with 31-68% in women. Among these, most had wedge deformities of the thoracic spine. On the other hand, up to 80% of osteoporotic vertebral fractures in men and up to 48% in women were missed by morphometry, in particular endplate fractures at the lumbar spine. In the group with osteoporotic vertebral fractures by RDC the proportion of persons with osteoporosis according to the WHO criteria (T-score < -2.5 SD) was 90.0% in women and 86.6% in men, compared with 67.9-85.0% in women and 20.8-50.0% in men with vertebral deformities by various methods. Although vertebral deformities by most definitions were significantly and inversely related to BMD as a continuous variable in both sexes [OR; 95% CI ranged between (1.70; 1.07-2.70) and (3.69; 1.33-10.25)], a much stronger association existed between BMD and osteoporotic fractures defined by RDC [OR; 95% CI between (4.85; 2.30-10.24) and (15.40; 4.65-51.02)]. In the nonosteoporotic group individuals with severe spondylosis had significantly higher BMD values at the femoral neck (p < 0.01) and lumbar spine (p < 0.0004) compared with the normal group. On the basis of internal (RDC) and external (BMD) validation, we conclude that assessment of vertebral osteoporotic fracture by quantitative methods alone will result in considerable misclassification, especially in men. Criteria for differential diagnosis as used within RDC can be helpful for a standardized subclassification of vertebral deformities in studies of spinal osteoporosis. PMID- 10793869 TI - Risk of hip fracture derived from relative risks: an analysis applied to the population of Sweden. AB - Bone mineral density measurements are widely used to estimate the relative risk of hip fracture. In addition, many other risk factors have been identified, some of which are known to add to the risk independently of other risk factors, including bone mineral density measurements. In this paper we develop an algorithm that converts relative risks for hip fracture to absolute (15 years and lifetime) risks, modeled on the population of Sweden. Lifetime risks increased as expected with increments in relative risk. Average lifetime risk in women at the age of 50 years was 22.7%, which increased to 64.9% when the relative risk was 6.0. In men the risk increased from 11.1% to 41.3%. The identification of high risk groups had little effect on the specificity of assessments but increased the sensitivity over a wide range of assumptions. The increment in lifetime risk was relatively stable across all ages, reducing the complexity of computing lifetime risks from relative risk. The derivation of absolute risk from relative risk permits the optimization of selection of individuals or populations either for further risk assessment or for treatment. PMID- 10793870 TI - Changes in levels of biochemical markers and ultrasound indices of Os calcis across the menopausal transition. AB - The aim of this longitudinal study was to investigate the changes in the levels of biochemical markers and ultrasound indices of os calcis across the menopausal transition. One hundred and ten healthy women (age 35-59 years at the 1992 baseline) participated in this 4-year population-based longitudinal study. Serum intact osteocalcin (IOC), urinary pyridinoline (Pyr), urinary deoxypyridinoline (Dpyr) and ultrasound indices were measured at baseline and after 4 years. The percentage changes in biochemical markers (% delta IOC, % delta Pyr and % delta Dpyr) and the percentage decreases in the ultrasound indices (% delta SOS, % delta BUA and % delta Stiffness) were calculated. The values of % delta IOC and % delta Dpyr in the perimenopausal subgroup (-4 to -3 years since menopause) and the values of % delta SOS and % delta Stiffness in the perimenopausal subgroup ( 2 to 0 years since menopause) were significantly higher than those in other groups. Pyr was significantly correlated with % delta SOS (r = -0.467, p < 0.01) and % delta Stiffness (r = -0.330, p < 0.05) and Dpyr was significantly correlated with % delta SOS (r = -0.390, p < 0.05), % delta BUA (r = -0.353, p < 0.05) and % delta Stiffness (r = -0.454, p < 0.05), while % delta IOC was significantly correlated with % delta SOS (r = -0.278, p < 0.05), % delta BUA (r = -0.369, p < 0.01) and % delta Stiffness (r = -0.383, p < 0.01) in the peri- and postmenopausal groups. These results indicate that the increase in bone turnover occurs 4 years before menopause. However, the correlations between biochemical markers and ultrasound indices were too low to allow prediction of bone change in the individual patient. PMID- 10793871 TI - Predictors of fractures in elderly women. AB - In a prospective study of 348 apparently healthy women, aged 70 years and over (mean 80.3 years), we examined bone mineral density (BMD), biochemical markers of bone metabolism, and some easily measurable predictors in relation to hip and osteoporotic fractures. In addition, we constructed risk profiles for hip and osteoporotic fractures. At baseline, BMD at both hips, using dual-energy X-ray absorptiometry, body height and body weight were measured. At the same time, serum and urine samples were obtained for biochemical analysis. Serum samples were analyzed for vitamin D metabolites, sex hormone binding globulin, serum intact parathyroid hormone, osteocalcin, alkaline phosphatase, phosphate, albumin, calcium and creatinine. In 2 h fasting urine, hydroxyproline, type I collagen crosslinked N-telopeptide (NTx) and calcium excretion were measured. Furthermore, easily measurable predictors, such as previous fracture, body mass index (BMI) and mobility were assessed. During the follow-up period (mean duration 5.0 years), hip and any osteoporotic fracture (wrist, humerus or hip fracture) occurred in 16 and 33 participants, respectively. Data were analyzed using Cox regression analysis. BMD of the trochanter (per 1 SD decrease) and previous fracture were most strongly associated with hip fractures (adjusted relative risk (RR) = 3.0, 95% confidence interval (CI): 1.4-6.6; RR = 4.2, 95% CI: 1.5-11.6, respectively) and osteoporotic fractures (RR = 1.8, 95% CI: 1.1 2.8; RR = 2.9, 95% CI: 1.5-5.7, respectively). Previous fracture, BMI and mobility were identified as easily measurable predictors for hip fractures, whereas previous fracture, use of loop diuretics and age were predictors for osteoporotic fractures in the risk profile model. The risk of fractures can be predicted with three easily measurable predictors. This study confirms the importance of previous fracture as a predictor for hip fractures and other fractures. It also shows that the use of loop diuretics is a predictor for osteoporotic fractures. PMID- 10793872 TI - Anorexia nervosa: slow regain of bone mass. AB - In a retrospective study of women aged 18-30 years, aimed at assessing factors associated with peak bone mass (PBM), 13 of 239 study cases reported having had anorexia nervosa. The mean total femoral and lumbar bone mineral density (BMD) values were not significantly lower in women who had had anorexia than in the pooled group (mean Z-scores of -0.60 and -0.48). Cases with less than 6 years since the anorexia had on average a present weight 5.7 kg less than their premorbid weights, while cases with more than 6 years since the eating disorder had an average weight 22.5 kg above their pre-morbid weights. The cases who had not regained their weight had BMD values significantly lower than the pooled material (mean Z-scores -1.15 and -0.9 in the lumbar spine and total femur respectively). Those who had regained their weight had BMD values as predicted from their present anthropometric data, while those who had not regained their weight had BMD values that were substantially below that predicted from their present weight. Anorexia nervosa seems to be associated with a low BMD which is even lower than that which can be predicted from the weight loss alone. This suggests that weight loss and other factors, such as menstrual dysfunction and estrogen deficiency, are independent and thus additive causes of bone loss in anorexia nervosa. Recovery of BMD seems slow, but the BMD may become as predicted from the anthropometric data after restoration of body weight and menses. The potential for recovery of BMD seems intact for several years after menarche. PMID- 10793873 TI - Breastfeeding in early life and bone mass in prepubertal children: a longitudinal study. AB - The aim of this study was to determine whether breastfeeding in early life is associated with bone mass in prepubertal children. We studied 330 8-year-old male and female children from Southern Tasmania representing 47% of those who originally took part in a birth cohort study of risk factors for Sudden Infant Death Syndrome in 1988. Breastfeeding intention and habit were assessed in both 1988 and 1996. Bone mineral density was measured by dual-energy X-ray densitometry. Children who were breastfed had higher bone mineral density at the femoral neck (+0.20 SD, p = 0.07), lumbar spine (+0.25 SD, p = 0.03) and total body (+0.29 SD, p = 0.006) compared with those who were bottle-fed. This association with breastfeeding was present in children born at term (femoral neck: +0.26 SD, p = 0.05; lumbar spine: +0.34 SD, p = 0.007; total body: +0.41 SD, p = 0.0008) but not those born preterm, and remained significant after adjustment for size, lifestyle factors and socioeconomic factors. Breastfeeding for less than 3 months was not associated with increased bone mass at any site. In conclusion, this study has demonstrated a beneficial association between breastfeeding in early life and bone mass in 8-year-old children born at term, particularly those breastfed for 3 months or longer, which appears biological. If this association is confirmed in other populations and persists until the attainment of peak bone mass then the implication would be that osteoporosis prevention programs need to start very early in the life cycle. PMID- 10793874 TI - Do men and women fracture bones at similar bone densities? AB - When the World Health Organization (WHO) guidelines for the definition of osteoporosis in postmenopausal women were identified similar proposals were not developed for men as there was insufficient evidence about the relationship between bone density and fracture in men. We have therefore examined the relationship between bone density and vertebral fracture in men and women attending for assessment of possible osteoporosis. Two hundred and sixty-four women (age 64 [SD 10] years) and 37 men (age 55 [10] years) were studied. Bone density was measured in the lumbar spine and femoral neck by dual-energy X-ray absorptiometry and expressed both as bone mineral density (BMD; g/cm2) and as T scores. In both sexes there was a sigmoid relationship between the cumulative frequency of vertebral fracture and bone density at both sites. There was a linear relationship between the log odds of fracture and bone mass for both sexes and both sites (r = 0.97-0.99; p < 0.0001). The slope of these lines was significantly steeper for men than women. The BMD at which there was 50% risk of fracture was higher in men than women (0.908 vs 0.844 g/cm2). The difference between the slopes was similar when the bone mass was expressed as a T-score. However, the T-score associated with 50% prevalence of fracture was similar in the two sexes (F: -2.77 vs M: -2.60). We conclude that although there is a different relationship between bone density and fracture in the two sexes the current WHO definition of osteoporosis in postmenopausal women can be appropriately applied to men. PMID- 10793875 TI - Axial and total-body bone densitometry using a narrow-angle fan-beam. AB - We assessed a new dual-energy bone densitometer, the PRODIGY, that uses a narrow angle fan-beam (4.5 degrees) oriented parallel to the longitudinal axis of the body (i.e., perpendicular to the usual orientation). High-resolution scans across the body can be stepped at 17 mm intervals. The energy-sensitive array detector uses cadmium zinc telluride, which allowed rapid photon counting. Spine and femur scans required 30 s, and total-body scans required 4-5 min; the dose was only 3.7 mrem and 0.04 mrem respectively, or about 5 to 10 times lower than conventional fan-beam densitometry. We found only a small influence of soft-tissue thickness on bone mineral density (BMD) results. There was also a small (+/- 1%) influence of height above the tabletop on BMD results. A software correction for object height allowed a first-order correction for the large magnification effects of position on bone mineral content (BMC) and area. Consequently, the results for BMC and area, as well as BMD, with PRODIGY corresponded closely to those obtained using the predecessor DPX densitometer, both in vitro and in vivo; there was a generally high correlation (r = 0.98-0.99) for BMD values. Spine and femur values for BMC, area and BMD averaged within 0.5% in vivo (n = 122), as did total-body BMC and BMD (n = 46). PRODIGY values for total-body lean tissue and fat also corresponded within 1% to DPX values. Regional and total-body BMD were measured with 0.5% precision in vitro and 1% precision in vivo. The new PRODIGY densitometer appears to combine the low dose and high accuracy of pencil-beam densitometry with the speed of fan-beam densitometers. PMID- 10793876 TI - Femur BMD bias associated with DPX-IQ reanalysis of DPX-acquired scan files. AB - Bone mineral density values calculated by DPX-IQ software reanalysis of original DPX software analyzed scans were compared at the femur region. Based on the results for 24 subjects each scanned on 12 different occasions, a significant bias at all sites--neck, Ward's triangle and trochanter--ranging from -0.7% (neck) to -2.1% (trochanter) was found. For the trochanter site 16 of the 24 subjects had significantly lower mean BMD values after DPX-IQ reanalysis, a trend also noted at the neck and Ward's triangle. Although it has been recommended that scans analyzed with DPX software do not require DPX-IQ reanalysis, our findings strongly indicate that DPX-IQ reanalysis is essential to minimize total error during follow-up BMD determinations. PMID- 10793877 TI - Intermittent oral disodium pamidronate in established osteoporosis: a 2 year double-masked placebo-controlled study of efficacy and safety. AB - The effect of oral pamidronate on bone mineral density and its adverse effect profile was investigated by a double-masked placebo-controlled study of 122 patients aged 55-75 years with established vertebral osteoporosis. Patients on active therapy received disodium pamidronate 300 mg/day (group A) for 4 weeks every 16 weeks, 150 mg/day (group B) for 4 weeks every 8 weeks or placebo (group C). All patients additionally received 500 mg of calcium and 400 IU vitamin D daily. Dual-energy X-ray absorptiometry measurements of the spine, hip, forearm and total body were performed at baseline and 6-monthly for 2 years using a Hologic QDR 1000 device at two sites. Serum osteocalcin and urinary deoxypyridinoline were measured at the above visits and at 3 months. The percentage change (SEM) in spine bone mineral density (BMD) at 2 years based on intention-to-treat analysis was 4.64 (1.01) in group A, 6.10 (0.87) in group B and 1.13 (1.32) in group C. Analysis of variance showed significant increases in group A and B compared with placebo (p < 0.01). There were also significant rises in femoral neck BMD for group A (p = 0.005), trochanter BMD for groups A and B (p < 0.01) and total-body BMD for groups A and B (p < 0.001). There was a significant reduction in serum osteocalcin and urinary deoxypyridinoline for groups A and B (p < 0.01). There was an excess of gastrointestinal side-effects in the treated groups, particularly group A. We conclude that intermittent pamidronate therapy can prevent bone loss at both the lumbar spine and femoral neck in patients with established vertebral osteoporosis, although due to gastrointestinal side-effects the 300 mg dose in particular does not appear suitable for clinical usage. PMID- 10793879 TI - Progress and paradigm shifts in vision research. PMID- 10793878 TI - A combination of low doses of 17 beta-estradiol and norethisterone acetate prevents bone loss and normalizes bone turnover in postmenopausal women. AB - The effects of 17 beta-estradiol (E2) 1 mg combined with low doses of norethisterone acetate (NETA) on postmenopausal bone loss and turnover were investigated in a 2-year, randomized, double-masked, placebo-controlled trial. A total of 135 postmenopausal women with a lumbar spine bone mineral density (BMD) T-score between -2 and +2 were randomized to daily treatment with an oral tablet of either placebo, E2 1 mg/NETA 0.25 mg, or E2 1 mg/NETA 0.5 mg. Significant (p < 0.001) increases in BMD at the lumbar spine (L1-4) were observed with E2 1 mg/NETA 0.25 mg (5.2%) and E2 1 mg/NETA 0.5 mg (5.4%) compared with placebo ( 0.9%). The total hip BMD increased significantly in the E2 1 mg/NETA 0.25 mg (3.1%) and E2 1 mg/NETA 0.5 mg groups (3.3%) compared with placebo. At the femoral trochanter, the increase in BMD in the E2 1 mg/NETA 0.5 mg group (6.3%) was significantly different from the placebo group (0.8%), while that in the E2 1 mg/NETA 0.25 mg group (3.3%) was not. No statistical differences were found between the active groups and placebo for the change in BMD at the femoral neck. Significant increases in BMD at the distal radius and total body were found for both E2 1 mg/NETA 0.25 mg (0.9% and 2.5%, respectively) and E2 1 mg/NETA 0.5 mg (2.1% and 3.0%, respectively) compared with placebo (-0.7% and 0.4%, respectively). At the end of the treatment, urinary pyridinoline type I collagen C-telopeptide had decreased by 65% and 60% in the E2 1 mg/NETA 0.25 mg and E2 1 mg/NETA 0.5 mg groups, respectively, while the mean serum concentrations of osteocalcin had decreased by 39% and 34%, bone-specific alkaline phosphatase by 32% and 29%, and C-terminal propeptide of type I collagen by 21% and 19% had decreased by 34-39%, 29-32%, and 19-21% in the E2 1 mg/NETA 0.25 mg and E2 1 mg/NETA 0.25 mg groups, respectively. In conclusion, combinations of E2 1 mg and NETA 0.25 or 0.5 mg prevent bone loss in postmenopausal women at the lumbar spine, hip, distal radius and total body, and normalize bone turnover. PMID- 10793880 TI - Trends in European visual science: 1978-1997. AB - European visual science during the past 20 years--with the glaring exception of ophthalmological research--is reflected in the annual programmes of the European Conference on Visual Perception. The number of countries that have participated has increased with time, and the spectrum of topics that have been presented has broadened. The number of multiple-author papers has increased dramatically, as has the number of papers by authors from different institutions and different countries. PMID- 10793881 TI - Progress and paradigm shifts in spatial vision over the 20 years of ECVP. AB - In the beginning there was light, and form, and visual mechanisms. This paper traces developments in research on spatial vision over the 20 years of ECVP, with particular emphasis on (1) hyperacuity, (2) peripheral vision, (3) amblyopia and development, and (4) learning and plasticity. PMID- 10793882 TI - From elements to perception: local and global processing in visual neurons. AB - Gestalt psychologists in the early part of the century challenged psychophysical notions that perceptual phenomena can be understood from a punctate (atomistic) analysis of the elements present in the stimulus. Their ideas slowed later attempts to explain vision in terms of single-cell recordings from individual neurons. A rapprochement between Gestalt phenomenology and neurophysiology seemed unlikely when the first ECVP was held in Marburg, Germany, in 1978. Since that time, response properties of neurons have been discovered that invite an interpretation of visual phenomena (including illusions) in terms of neuronal processing by long-range interactions, as first proposed by Mach and Hering in the last century. This article traces a personal journey into the early days of neurophysiological vision research to illustrate the progress that has taken place from the first attempts to correlate single-cell responses with visual perceptions. Whereas initially the receptive-field properties of individual classes of cells--e.g., contrast, wavelength, orientation, motion, disparity, and spatial-frequency detectors--were used to account for relatively simple visual phenomena, nowadays complex perceptions are interpreted in terms of long-range interactions, involving many neurons. This change in paradigm from local to global processing was made possible by recent findings, in the cortex, on horizontal interactions and backward propagation (feedback loops) in addition to classical feedforward processing. These mechanisms are exemplified by studies of the tilt effect and tilt aftereffect, direction-specific motion adaptation, illusory contours, filling-in and fading, figure--ground segregation by orientation and motion contrast, and pop-out in dynamic visual-noise patterns. Major questions for future research and a discussion of their epistemological implications conclude the article. PMID- 10793883 TI - Single units and sensation: a retrospect. AB - The single-neuron doctrine is reexamined, and the search for causal links between single units and sensation reviewed. Although several decades of single-unit recording have been very successful in elucidating physiological mechanisms, linking signals from a single cell and perception has progressed at a slower rate. Nevertheless, analysing the activity of single neurons has achieved significant gains and remains the most promising level for elucidation of processing algorithms in the visual system. At the subcortical level, the conclusion that signals from just a single cell can carry enough information for some kinds of performance remains (almost) valid. Under carefully controlled conditions, just a few impulses in a few retinal ganglion cells are an adequate signal for the cortex to initiate a behavioural response. Elucidating cortical codes has been more difficult, and evidence exists suggesting the sharing of responsibility for a task among cell assemblies; how large these assemblies are, and how to test for them neurophysiologically, remains a challenge. PMID- 10793884 TI - Paradigm shifts: new techniques to answer new questions. AB - The advent of a multiple-channels approach to spatial vision 20 years ago raised important questions that were difficult to approach empirically, given the technology and analytic tools of the time. These questions concerned the interaction or combination of different components of a stimulus--questions that have recently resurfaced in more complex form. Classical psychophysical methods for assessing whether two stimulus aspects are coded independently (e.g., masking, adaptation, and cue-summation) provide only limited information about the nature of whatever interactions are discovered. In both older work in detection and recent work in complex pattern discrimination, we have used a double-judgment paradigm in which the observer rates two aspects of a stimulus simultaneously. The paradigm provides a rich source of information about the codes underlying each psychophysical decision and which are unique in permitting us to psychophysically investigate effects resulting from neural noise in the system. Our analyses draw on theories of dimensional interaction in signal detection theory and in information theory, and on methods from several branches of statistics, including categorical data analysis and structural equation modeling. We review the theoretical, technological, methodological, and personal influences that led us to develop this approach. PMID- 10793885 TI - Visual perception in 'low vision'. AB - 'Low vision' has an international classification as an impairment of vision that is defined by loss of visual acuity and the size of the visual field. Functional classification that would depict capability to use vision in different activities has not yet been agreed upon. Abnormal visual information leads to changes in visual perception, and brain-damage-related visual impairment may alter or prevent one or several cognitive visual functions. Abnormal visual perception opens a window into brain mechanisms that the psychophysicists otherwise do not have. Together with clinicians, psychophysicists can help visually impaired persons to understand the nature of their disability by thoroughly studying the nature of the altered image and the profile of cognitive visual functions. PMID- 10793886 TI - The generation of lipid signaling molecules in the nucleus. PMID- 10793887 TI - Vitamin E and its function in membranes. AB - Vitamin E is a fat-soluble vitamin. It is comprised of a family of hydrocarbon compounds characterised by a chromanol ring with a phytol side chain referred to as tocopherols and tocotrienols. Tocopherols possess a saturated phytol side chain whereas the side chain of tocotrienols have three unsaturated residues. Isomers of these compounds are distinguished by the number and arrangement of methyl substituents attached to the chromanol ring. The predominant isomer found in the body is alpha-tocopherol, which has three methyl groups in addition to the hydroxyl group attached to the benzene ring. The diet of animals is comprised of different proportions of tocopherol isomers and specific alpha-tocopherol-binding proteins are responsible for retention of this isomer in the cells and tissues of the body. Because of the lipophilic properties of the vitamin it partitions into lipid storage organelles and cell membranes. It is, therefore, widely distributed in throughout the body. Subcellular distribution of alpha-tocopherol is not uniform with lysosomes being particularly enriched in the vitamin compared to other subcellular membranes. Vitamin E is believed to be involved in a variety of physiological and biochemical functions. The molecular mechanism of these functions is believed to be mediated by either the antioxidant action of the vitamin or by its action as a membrane stabiliser. alpha-Tocopherol is an efficient scavenger of lipid peroxyl radicals and, hence, it is able to break peroxyl chain propagation reactions. The unpaired electron of the tocopheroxyl radical thus formed tends to be delocalised rendering the radical more stable. The radical form may be converted back to alpha-tocopherol in redox cycle reactions involving coenzyme Q. The regeneration of alpha-tocopherol from its tocopheroxyloxyl radical greatly enhances the turnover efficiency of alpha tocopherol in its role as a lipid antioxidant. Vitamin E forms complexes with the lysophospholipids and free fatty acids liberated by the action of membrane lipid hydrolysis. Both these products form 1:1 stoichiometric complexes with vitamin E and as a consequence the overall balance of hydrophobic:hydrophillic affinity within the membrane is restored. In this way, vitamin E is thought to negate the detergent-like properties of the hydrolytic products that would otherwise disrupt membrane stability. The location and arrangement of vitamin E in biological membranes is presently unknown. There is, however, a considerable body of information available from studies of model membrane systems consisting of phospholipids dispersed in aqueous systems. From such studies using a variety of biophysical methods, it has been shown that alpha-tocopherol intercalates into phospholipid bilayers with the long axis of the molecule oriented parallel to the lipid hydrocarbon chains. The molecule is able to rotate about its long axis and diffuse laterally within fluid lipid bilayers. The vitamin does not distribute randomly throughout phospholipid bilayers but forms complexes of defined stoichiometry which coexist with bilayers of pure phospholipid. alpha-Tocopherol preferentially forms complexes with phosphatidylethanolamines rather than phosphatidylcholines, and such complexes more readily form nonlamellar structures. The fact that alpha-tocopherol does not distribute randomly throughout bilayers of phospholipid and tends to form nonbilayer complexes with phosphatidylethanolamines would be expected to reduce the efficiency of the vitamin in its action as a lipid antioxidant and to destabilise rather than stabilise membranes. The apparent disparity between putative functions of vitamin E in biological membranes and the behaviour in model membranes will need to be reconciled. PMID- 10793888 TI - Integration of phospholipid and sterol metabolism in mammalian cells. AB - The lethal consequences of imbalances in lipid and sterol metabolism in human diseases such as atherosclerosis and lipid storage disorders underscores our need to know how cholesterol, phospholipid and sphingolipid metabolism is integrated. Accumulation and abnormal localization of lipids and sterol affects cellular function not only by perturbing membrane activity but also by increasing production of bioactive lipids derived from cholesterol, phospholipids and sphingolipids. For example in the NPC mouse model, accumulation of intracellular cholesterol and sphingomyelin is accompanied by increased sphingosine [187], a potent regular of protein kinase C and cell proliferation [152]. Oxidized LDL has an important role in the pathology of atherosclerosis by promoting foam cell formation and cytotoxicity [65]. 7-Hydroxycholesterol and 7-ketocholesterol are involved in many aspects of oxidized LDL activity including initiation of apoptosis in a number of cell types [188, 189] and enhancing cholesterol accumulation by inhibiting efflux [190]. Oxysterols formed intracellularly or from oxidized lipoproteins could have an important role in regulating lipid metabolism in the foam cell. Bioactive metabolites of phospholipids, such as diglyceride, phosphatidic acid and lysolipids, could also increase in circumstances of elevated deposition and have profound and varied effects on cell physiology. In addition to elucidating mechanisms for integration of lipid metabolism, we should determine when these responses go awry and assess the influence of bioactive compounds formed under these circumstances on cell viability and growth. PMID- 10793889 TI - Phospholipid biosynthesis in the yeast Saccharomyces cerevisiae and interrelationship with other metabolic processes. AB - In this review, we have discussed recent progress in the study of the regulation that controls phospholipid metabolism in S. cerevisiae. This regulation occurs on multiple levels and is tightly integrated with a large number of other cellular processes and related metabolic and signal transduction pathways. Progress in deciphering this complex regulation has been very rapid in the last few years, aided by the availability of the sequence of the entire Saccharomyces genome. The assignment of functions to the remaining unassigned open reading frames, as well as ascertainment of remaining gene-enzyme relationships in phospholipid biosynthesis in yeast, promises to provide detailed understanding of the genetic regulation of a crucial area of metabolism in a key eukaryotic model system. Since the processes of lipid metabolism, secretion, and signal transduction show fundamental similarities in all eukaryotes, the dissection of this regulation in yeast promises to have wide application to our understanding of metabolic control in all eukaryotes. PMID- 10793890 TI - The study of storage lipid biosynthesis using microspore-derived cultures of oil seed rape. PMID- 10793891 TI - Acyltransferases of de novo glycerophospholipid biosynthesis. PMID- 10793892 TI - Illusory spatial offset of a flash relative to a moving stimulus is caused by differential latencies for moving and flashed stimuli. AB - A flash that is presented adjacent to a continuously moving bar is perceived to lag behind the bar. One explanation for this phenomenon is that there is a difference in the persistence of the flash and the bar. Another explanation is that the visual system compensates for the neural delays of processing visual motion information, such as the moving bar, by spatially extrapolating the bar's perceived location forward in space along its expected trajectory. Two experiments demonstrate that neither of these models is tenable. The first experiment masked the flash one video frame after its presentation. The flash was still perceived to lag behind the bar, suggesting that a difference in the persistence of the flash and bar, does not cause the apparent offset. The second experiment employed unpredictable changes in the velocity of the bar including an abrupt reversal, disappearance, acceleration, and deceleration. If the extrapolation model held, the bar would continue to be extrapolated in accordance with its initial velocity until the moment of an abrupt velocity change. The results were inconsistent with this prediction, suggesting that there is little or no spatial compensation for the neural delays of processing moving objects. The results support a new model of temporal facilitation for moving objects whereby the apparent flash lag is due to a latency advantage for moving over flashed stimuli. PMID- 10793893 TI - Unconstrained stereoscopic matching of lines. AB - The computation of horizontal binocular disparities used in stereoscopic depth perception depends upon the identification of corresponding features in the two retinal images. In principle, binocular matching is a two-dimensional problem that considers matches in all possible meridians. Normally, constraints such as end points or crossing points limit the direction and magnitude of matches. If matching is unconstrained, such as is the case with long lines, it is completely ambiguous. Under these conditions the default match will be determined by the operating range, or upper disparity limit, of matchable vertical and horizontal disparities. We computed the operating range of vertical matches for stereoscopic depth as a function of line orientation. Our results suggest that the two dimensional operating range is anisotropic for vertical and horizontal disparity and that unconstrained matches are not based upon either epipolar geometry or nearest neighbor constraints, but rather the mean of disparity estimates within the operating range for binocular matches. This operating range can be extended vertically when matches are constrained by image primitives. PMID- 10793894 TI - Spatial scale of visual analysis for vernier acuity does not vary over time. AB - The visual system filters spatial pattern through a range of narrowly tuned spatial mechanisms, but the rules by which the outputs of these mechanisms are combined across time to extract precise geometrical information are not yet clear. One hypothesis is that spatial analysis shifts over time from coarse to finer spatial scales, in order to extract fine spatial information. An alternative hypothesis is that thresholds are determined by the signal-to-noise ratio within an optimal spatial scale. In this study, we measured vernier acuity across exposure duration for equally visible long lines and short lines and found no improvement in spatial precision with time. Using a simultaneous spatial masking paradigm, we determined the active spatial scales at 100 and 1000 ms. The results show no significant changes in spatial scale, or in the size-range of active scales, for the two exposure durations. Furthermore, whereas vernier thresholds vary markedly with line contrast, we find only modest shifts in spatial scale. Taken together, our results suggest that for vernier acuity, spatial scale is selected very early, and that vernier thresholds are predominantly limited by signal strength within that spatial scale. PMID- 10793895 TI - Attention mechanisms for multi-location first- and second-order motion perception. AB - We applied the external noise plus attention paradigm to study attention mechanisms involved in concurrent first-order and second-order motion perception at two spatial locations. Cued to attend to one of the locations, the observer was instructed to independently judge direction of motion of either first-order (Experiment 1) or second-order (Experiment 2) motion stimuli at both locations in every trial. Across trials, systematically controlled amounts of external noise were added to the motion displays. We measured motion threshold at three performance criteria in every attention x external noise condition. We find that observers could, without any loss, simultaneously compute first-order motion direction at two widely separated spatial locations across a broad range of external noise conditions. However, considerable loss occurred at the unattended location in processing second-order motion direction at two separated spatial locations. We conclude that, under the conditions investigated in the current study, (1) in first-order motion perception, the visual system could simultaneously process motion direction at two widely separated locations without any capacity limitation; (2) in second-order motion perception, attending to a spatial location enhances stimulus contrast at that location by a factor of about 1.37 (or equivalently, reduces the internal additive noise by a factor of about 0.73). PMID- 10793896 TI - Motion detection from photopic to low scotopic luminance levels. AB - In this study we quantify the influence of adaptation luminance on the threshold for direction-detection in coherently moving random-pixel arrays (RPAs). Square RPAs of a constant rms-contrast (35%) were used and we determined their 'critical' or threshold-width Wc. Mean retinal illuminances were varied in 13 steps of 0.5 log unit from the low photopic range (screen luminance 0.3 cd/m2) down to 6 log units attenuation, which appeared to be about the absolute threshold of vision under the conditions of our experiment. Moving RPAs were presented at six retinal locations (0, 3, 6, 12, 24 and 48 degrees) from the fovea to the far periphery in the temporal visual field of the right eye of three experienced observers (the authors). In order to ensure an honest comparison between these very disparate conditions, the spatial dimensions (including speed) were scaled according to the acuity, as measured separately for each of the viewing-conditions and observers. Acuity scaling proves to equate the performance for all eccentricities and luminance levels rather well. The fovea is special, but only in the sense that the absolute threshold for light detection is reahed earlier than in peripheral regions. In all other respects foveal results follow the pattern found for peripheral locations. Two different regimes can be discerned in the data, one for high and one for low speeds. In the low speed range Wc is almost constant, regardless of luminance level or eccentricity. The critical 'crossing-time' Tc for any pixel starting at one end of the stimulus and leaving at the opposite end is therefore inversely proportional to velocity in the low-speed range (time-velocity reciprocity). At medium-to-high speeds Wc increases linearly with velocity, so Tc is constant. This constant (minimum) value of Tc differs between subjects, but in all subjects it increases somewhat with decreasing luminance level, even for our acuity-scaled stimuli. The different behaviour for low and high speeds [reported before for photopic viewing conditions by van de Grind, W. A., van Doorn, A. J., & Koenderink, J. J. (1983. Journal of the Optical Society of America, 73, 1674-1683) and van de Grind, W. A., Koenderink, J. J., & van Doorn A. J. (1986. Vision Research, 26, 797-810)] proves to hold from photopic to low scotopic luminance ranges, provided the stimuli are scaled according to acuity. We draw the general conclusion that movement detection is a very robust process that tolerates extremely low retinal illuminance levels. Moreover, the visual system appears to use the same processing principles in combination with an acuity-scaled architecture under all adaptation states and at all eccentricities. PMID- 10793897 TI - A model of the perceived relative positions of moving objects based upon a slow averaging process. AB - We extend the local energy model of position detection to cope with temporally varying position signals and the perception of relative position. The extension entails two main components. First, a form of persistence for the position signal based on the temporal impulse response function of the visual system. Secondly, we hypothesise that the perceived relative position of two objects is determined by a slow average of the difference of the objects' position signals. The model explains why briefly flashed static dots are perceived to lag behind continuously visible moving dots, without the need for a motion extrapolation process [Nijhawan, R. (1994). Nature, 370, 256-257]. The dependence of this illusion on parameters such as the velocity, duration, frequency and number of flashes of the motion trajectories is accurately captured by the model. Furthermore, the model makes two predictions. First, briefly flashed dots on a staircase trajectory should lead dots with a long duration. Secondly, it should be possible to abolish the lag-effect between continuously visible and stroboscopically moving objects by halting the continuously visible dots during the interflash interval of the stroboscopic dots. Both predictions are corroborated in experiments. PMID- 10793898 TI - Perception of three-dimensional shape from texture is based on patterns of oriented energy. AB - This paper presents empirical support for a new observer model of inferring three dimensional shape from monocular texture cues. By measuring observers' abilities to estimate the relative three-dimensional curvature along a textured surface from two-dimensional projected images, and concurrently examining the local spectral changes occurring in the projected image for various texture patterns, we have found that correlated changes in oriented energy along lines corresponding to the lines of maximum and minimum curvature of the surface are crucial for conveying the three-dimensional shape of the surface. Energy along these lines of maximum and minimum curvature can be used to compute the orientation of local surface patches. Texture patterns consisting of simple and complex sinusoidal gratings and plaids, and filtered noise were drawn onto a surface that was corrugated sinusoidally in depth about the horizontal axis and projected in perspective onto an image plane. The perceived relative surface curvature was reconstructed from measurements of local ordinal depth around a central fixation point at 12 different phases of the corrugation. Our results show that: (1) it is neither necessary nor sufficient to identify individual texture elements or texture gradients in order to extract the shape of the surface; (2) one-dimensional frequency modulation is insufficient for conveying complex three-dimensional shape. (3) Veridical ordinal depth is seen only when the projected pattern contains changes in oriented energy along lines corresponding to projected lines of maximum curvature of the surface. (4) For a surface corrugated in depth about the horizontal axis, this pattern of oriented energy arises from energy along the vertical direction in the global Fourier transform of the pre-corrugated pattern. (5) Local orientation changes across lines of minimum curvature can be also critical for conveying shape. (6) These correlated orientation changes along lines of maximum and minimum curvature are entirely lost in parallel projection. Hence texture is a useful cue for shape if the image is a perspective projection. (7) Only some natural textures will provide sufficient monocular cues to support veridical shape inferences, and this can be predicted from their global Fourier transforms. PMID- 10793899 TI - A model of human heading judgement in forward motion. AB - We developed a new computational model of human heading judgement from retinal flow. The model uses two assumptions: a large number of sampling points in the flow field and a symmetric sampling region around the origin. The algorithm estimates self-rotation parameters by calculating statistics whose expectations correspond to the rotation parameters. After the rotational components are removed from the retinal flow, the heading direction is recovered from the flow field. Performance of the model was compared with human data in three psychophysical experiments. In the first experiment, we generated stimuli which simulated self-motion toward the ground, a cloud or a frontoparallel plane and found that the simulation results of the model were consistent with human performance. In the second and third experiment, we measured the slope of the perceived versus simulated heading function when a perturbation velocity weighted according to the distance relative to the fixation distance was added to the vertical velocity component under the cloud condition. It was found that as the magnitude of the perturbation was increased, the slope of the function increased. The characteristics observed in the experiments can be explained well by the proposed model. PMID- 10793900 TI - A hierarchical structure of motion system revealed by interocular transfer of flicker motion aftereffects. AB - Interocular transfer of the motion aftereffect (MAE) has been extensively investigated for the purpose of analysing the binocularity of the underlying motion mechanism. Previous studies unanimously reported that the transfer of the classical static MAE is partial, but there is a controversy as to whether the transfer of the flicker MAE (MAE measured using counterphase gratings) is partial or perfect. To gain insight into the discrepancy between studies, we investigated whether the interocular transfer of the flicker MAE is influenced by the MAE measurement method, retinal eccentricity and attention. Our results showed that the transfer was perfect or nearly so when the MAE duration was measured in the central visual field with observers paying attention to the adaptation stimulus, but the transfer was partial when the MAE nulling strength was measured, when the MAE duration was measured in the peripheral visual field, or when the observers' attention was distracted by a secondary task. These results not only resolve discrepancies between previous studies, but also suggest that the flicker MAE reflects adaptation at multiple stages in the hierarchical architecture of motion processing. PMID- 10793902 TI - Visual memory for random block patterns defined by luminance and color contrast. AB - We studied the ability of human subjects to memorize the visual information in computer-generated random block patterns defined either by luminance contrast, by color contrast, or by both. Memory performance declines rapidly with increasing inter-stimulus interval, showing a half-life of approximately 3 s. We further show that memory performance declines with eccentricity approximately as a Gaussian function of position. Memory decay functions did not depend on whether the patterns were defined by luminance or color contrast. Changing both luminance and color components of block patterns in conjunction did not improve performance suggesting a single memory mechanism is used to store luminance and color derived pattern information. Our results further suggest that color identity (hue, saturation) and pattern information extracted from color- or luminance-contrast are stored independently of each other. PMID- 10793901 TI - Facilitation from collinear flanks is cancelled by non-collinear flanks. AB - Detection of a central Gabor pattern is facilitated by the presence of collinear flanking patterns. We find that this facilitation is greatly reduced when the collinear flanks are combined with non-collinear flanks to form a coherent surround. These results are unlikely to be explained by mechanisms that merely transduce local contrast in a nonlinear fashion. A model wherein the outputs of such mechanisms are combined anisotropically provides a better account for these results. PMID- 10793903 TI - Motion and shape in common fate. AB - We determined how much motion coherence was needed to detect a target group of four moving dots in a dynamic visual noise (DVN) background. The lifetimes of the trajectories of the target and that of the noise dots were the same. In addition to parallel trajectories and collinear dot arrangements, divergent, convergent, or crossing trajectories and non-collinear dot arrangements were also tested. Performance saturated at a lifetime of approximately 600 ms. It was best for parallel trajectories and collinear dots, and worse for crossed trajectories with non-collinear dots, where it approached performance in a no-motion, form-only control experiment. Results illustrate the importance of common fate in motion perception in DVN, when other factors are equated. PMID- 10793904 TI - Sensitivity to contrast modulation depends on carrier spatial frequency and orientation. AB - We consider how the detection of second-order contrast structure depends on the orientation and spatial frequency of first-order luminance structure. For patterns composed of a bandpass noise carrier multiplied by a contrast envelope function, we show that sensitivity to the envelope varies in proportion to the spatial frequency of the carrier. For oriented carriers at low spatial frequencies, detection of the contrast envelope is easier when the envelope and carrier are perpendicular, but this dependency diminishes as the spatial frequency of the carrier increases. These differences are not attributable to either the detection of side-bands, or the presence of spurious contrast structure in unmodulated carrier images. A final experiment measured envelope detection in the presence of noise masks. Results indicate that orientationally and spatially-band pass filtering precedes the detection of second-order structure. PMID- 10793905 TI - Cross-domain adaptation reveals that a common mechanism computes stereoscopic (cyclopean) and luminance plaid motion. AB - Across three experiments, this study investigated the visual processing of moving stereoscopic plaid patterns (plaids created with cyclopean components defined by moving binocular disparity embedded in a dynamic random-dot stereogram). Results showed that adaptation to a moving stereoscopic plaid or its components affected the perceived coherence of a luminance test plaid, and vice versa. Cross-domain adaptation suggests that stereoscopic and luminance motion signals feed into a common pattern-motion mechanism, consistent with the idea that stereoscopic motion signals are computed early in the motion processing stream. PMID- 10793906 TI - Disparity vergence double responses processed by internal error. AB - Disparity vergence eye movements occasionally exhibit two high-velocity components to a single step stimulus (Alvarez, T. L., Semmlow, J. L. & Yuan, W. (1998). Journal of Neurophysiology, 79, 37-44). This research investigates the neural strategy used to trigger the second component of double high-velocity vergence eye movements. Vergence doubles evoked by an experimental protocol that induces post-movement visual error were compared to doubles that occur normally. The second component of a visually evoked response double occurred later, and with slower dynamics, than that of a naturally occurring double. These differences in timing and dynamics indicate that natural double responses are mediated, at least in part, by a mechanism other than visual feedback. The faster dynamics and timing of natural doubles suggest that an internal monitoring process triggers these movements. PMID- 10793907 TI - Tilt dependency of slant aftereffect. AB - Slant aftereffect (SAE), the negative aftereffect of slant induced after prolonged observation of a surface, is considered as evidence that slant is encoded in the visual system. Because slant and tilt are mathematically independent dimensions, Stevens (Stevens, K. A. (1983a). Biological Cybernetics, 46, 183-195) assumed that slant and tilt are processed independently in the visual system. To confirm this assumption, we investigated whether SAE is induced independently of the difference in tilt between the adapting and test stimuli. The stimuli were displayed by simulating the motion disparity of rotating disks. After adaptation to a surface of 60 degrees slant, the subjective 0 degree slants of the test stimulus were measured with the tilt differences of 0, 45, 90, 135 and 180 degrees. The magnitude of SAE was greatest when the tilt difference was zero, and decreased with increasing tilt difference. The results suggest that slant and tilt are not processed independently in the visual system and that the slant detector in the visual system is sensitive not only to slant but also to tilt. PMID- 10793908 TI - Toxicity of drugs used for treatment of rhinitis: a reminder to the otorhinolaryngologist. AB - We retrospectively studied files on toxicity of drugs used for treatment of rhinitis (nasal antihistamines, oral antihistamines, nasal vasoconstrictors, oral decongestants, nasal corticosteroids, nasal anticholinergic agent, nasal mast cell stabilizer, and respiratory antigen preparations for oral use) that occurred in Sao Paulo, Brazil, from 1 July 1996 to 30 June 1998. There were 823 cases of toxicity by these agents, with a predominance of oral decongestants (in combination with antihistamines) (53.4%). There was a significant incidence of toxicity within children aged 1 to 4 years (P < 0.05) and a preponderance among males aged 5-9 years and females aged 10-39 years. Accidental intake of medicines accounted for the majority of toxicity cases (57.2%). PMID- 10793909 TI - The effect of head position on the distribution of drops within the nose. AB - Nose drops are commonly prescribed in the United Kingdom, yet clinicians vary considerably in the instructions they give on how to instill them. This is important because how they are used affects how well they work. In this study, a visual assessment of the intranasal distribution of drops was made using endoscopic examination after the instillation of betamethasone drops containing methylene blue. Drops instilled with the head tipped back were found along the nasal floor and in the nasopharynx, with none visible in the middle meatus or elsewhere on the lateral nasal wall. Drops instilled in Mygind's position or in the praying-to-Mecca position were found predominantly in the middle meatus and on the middle and inferior turbinates. No difference was seen between these two positions in the distribution of drops achieved, and this is likely to lead to similar clinical efficacy. Because Mygind's position is much more comfortable for patients, it should be recommended by all doctors prescribing nose drops. PMID- 10793910 TI - Proliferative activity and cytometric characteristics in polyps of the nasal cavity and paranasal sinuses. AB - Although several investigations have revealed the influence of cytokines, allergy, and environmental factors in polyp development, the etiology of nasal polyps is still unknown. To estimate the biology of this common disease the operative specimens of 50 patients who underwent surgery for polyps of the nasal cavity and the paranasal sinuses were examined; of these, 10 patients had recurrent disease and 23 patients had an allergy. The investigations included routine histology and quantitative DNA measurements, along with immunohistochemical identification of proliferation markers (i.e., MIB-1; proliferating cell nuclear antigen, PCNA). Histologically, most polyps revealed an infiltration with lymphocytes, eosinophilic granulocytes, and plasma cells. Twenty-five percent had a squamous metaplasia of the respiratory epithelium. Quantitative DNA analysis demonstrated diploid stemlines and lack of aneuploid cells with a DNA content exceeding 5c in most cases. Immunohistochemical detection of proliferation markers showed low proliferation rates in all cases. In 27 polyps no MIB-1 expression was detected, and in 7 polyps no PCNA expression was detected. The polyps of the 23 patients with proven allergic diathesis did not reveal higher scores for the parameters of DNA analysis (i.e., ploidy status and percentage of aneuploid cells) and proliferation scores. Nasal polyps of 10 patients with recurrent disease displayed higher scores for proliferation markers, and in five cases aneuploid cells with 5c exceeding rate (5cER) of 1.5 11.7% were detected. According to these results, polyps of the nasal cavity and paranasal sinuses showed low proliferation scores and were diploid. The data demonstrated that there was no increase of proliferation activity or ploidy shift toward aneuploidy in patients with allergy. Nevertheless, in recurrent disease some increase in proliferation activity and some changes in the parameters of the DNA analysis occurred, indicating more aggressive behavior of recurrent polyps in single cases. PMID- 10793911 TI - Identification of intranasal cerebrospinal fluid leaks by topical application with fluorescein dye. AB - The purpose of this paper is to describe a safe new technique for intraoperative identification of the site of cerebrospinal fluid rhinorrhea. Cerebrospinal fluid (CSF) rhinorrhea after intracranial or intranasal surgery is a known potential complication with significant morbidity and mortality. It is currently accepted that endoscopic intranasal management of CSF rhinorrhea is the preferred method of surgical repair, with higher success rates and less morbidity than intracranial surgical repair in selected cases. Accurate identification of the site of CSF leakage is necessary for a successful endoscopic surgical repair. Computer tomography (CT) with or without intrathecal contrast and preoperative nasal endoscopy are frequently used to preoperatively localize the site of the leak. Intrathecal fluorescein administered immediately before surgery has aided in the intraoperative identification of the site of CSF leak in 25-64% of patients undergoing endoscopic repair of CSF rhinorrhea in whom preoperative CT scanning and nasal endoscopy had not identified the site of CSF leak. Intrathecal fluorescein, however, has been associated with severe complications, such as lower extremity weakness, numbness, generalized seizures, opisthotonus, and cranial nerve deficits. We present three cases of CSF rhinorrhea in which fluorescein was applied intranasally during the endoscopic surgical repair. Ten percent fluorescein was applied to the nose with a cotton swab. Under endoscopic visualization the fluorescein changed its fluorescent color from amber/yellow to a dark green and was found streaming from high in the nasal cavity, which led to accurate identification of the site of the CSF leak. PMID- 10793912 TI - The use of acoustic rhinometry in predicting outcomes after sinonasal surgery. AB - Today's healthcare environment demands objective assessment of surgical outcomes. The recent otolaryngologic literature has established acoustic rhinometry (AR) as a valid instrument to objectively document nasal patency. The purpose of this article is to evaluate the utility of AR in predicting outcomes after sinonasal surgery. All patients scheduled for sinonasal surgery at the Tulane University and University of Mainz Departments of Otolaryngology between 10/1/98 and 12/15/98 were enrolled. All subjects underwent AR and completed a sinonasal outcome survey (SNOT-20) one day before and two months after their surgical procedure. Thirty-one patients were enrolled. The SNOT-20 raw scores improved from a mean of 7.93 (+/- 3.78) preoperatively to 3.35 (+/- 2.33) postoperatively (p < 0.05). The I-notch revealed no significant change postoperatively. The mean bilateral predecongestion C-notch increased from 1.257 cm2 to 1.451 cm2 (p < 0.05). Patients with a bilateral C-notch > 1 cm2 were 1.96 times more likely to have a five-point improvement in the SNOT-20 raw score (95% CI = 1.17, 3.27). The mean value of the C-notch is significantly altered (increased) as a result of sinonasal surgery. Patients with a preoperative cross-sectional area < 1 cm2 are less likely to report large postoperative subjective improvement. These results indicate that patients with poor geometry at the area of the C-notch do not fare as well surgically as those with better preoperative measurements. PMID- 10793913 TI - The bacteriology of chronic rhinosinusitis: results using a novel culture device. AB - The bacteriology of chronic rhinosinusitis is an area of great interest among rhinologists. Numerous studies have reported culture results implicating a variety of aerobic and anaerobic organisms in the etiology of this disease process. Sampling is generally accomplished through the nasal cavity, creating the potential for contamination with resident nasal flora. In some reports, strict anaerobic techniques have not been used, possibly accounting for the failure to recover these fastidious organisms. In an attempt to clarify the microbiology of chronic rhinosinusitis, we used a novel culture and transport system in 50 patients undergoing endoscopic sinus surgery. The Accu-CulShure (MLA Systems, Pleasantville, NY, USA) is a self-contained polyethylene culture swab and transport device, capable of collecting a representative sample from the sinus without contamination, and preserving the quality of the material during transport. Our aerobic and anaerobic culture results, as well as pertinent patient data, are presented. The Accu-CulShure device may permit standardization of culture techniques for future studies, and allow for more accurate determination of the microbiology of chronic rhinosinusitis. PMID- 10793914 TI - Endonasal endoscopic marsupialization of paranasal sinus mucoceles. AB - Paranasal sinus mucoceles are relatively common in Japan, especially after prior Caldwell-Luc surgery. Recently developed endonasal endoscopic approaches were used for the drainage and marsupialization of mucoceles. We present the surgical treatment of 97 patients with mucoceles, including 68 maxillary, 13 ethmoid, nine frontal, seven sphenoid, eight frontoethmoid, and one sphenoethmoid sinuses. Six patients with maxillary mucoceles were operated via a sublabial incision. For frontal mucoceles, four patients were treated by a combined external and endonasal endoscopic approach. The other mucoceles underwent complete marsupialization under endonasal endoscopic control. There were no intraoperative or postoperative complications. No evidence of recurrence was observed in any patients during follow-up periods ranging from 36 to 84 months. PMID- 10793915 TI - The endoscopic management of chronic frontal sinusitis associated with frontal sinus posterior table erosion. AB - Expansile inflammatory diseases of the frontal sinuses may produce erosion of the posterior table of the frontal sinus. In these instances, the bone between sinus mucosa and intracranial dura is absent. Over the past decade, endoscopic frontal sinusotomy has emerged as the preferred technique for the treatment of refractory chronic frontal sinusitis. Endoscopic approaches also have a role in the most advanced instances of frontal sinusitis. A retrospective chart review of patients who were treated for frontal sinusitis with erosion of the frontal sinus posterior table was performed. Eight patients were identified. All patients underwent endoscopic frontal sinusotomy; some patients required multiple endoscopic procedures. Complete frontal recess dissection with identification of the frontal ostium was achieved for all involved frontal sinuses. In all cases, this postoperative result was monitored by CT scans (where indicated) and serial nasal endoscopy, which demonstrated good frontal sinus aeration and normal mucociliary clearance. Antibiotics were administered for culture-documented bacterial exacerbations, and systemic steroids were given for management of allergic fungal sinusitis and sinonasal polyposis associated with asthma. No patient underwent frontal sinus obliteration or cranialization. No suppurative intracranial complications were noted during the postoperative period. Endoscopic frontal sinusotomy can be used safely for the definitive management of frontal sinusitis associated with posterior table erosion. In fact, endoscopic techniques may represent the preferred approach for the treatment of this problem. Such an approach avoids the morbidity of more destructive alternatives (such as obliteration), and serves to create a frontal sinus with normal mucociliary clearance. PMID- 10793916 TI - Nasal nitric oxide is not altered by topical anesthesia. AB - This prospective study was undertaken to determine whether topical nasal anesthetic agents affect nasal nitric oxide (NO) output in healthy adults. Seven volunteers (aged: 29-56 (40.6 +/- 10.7) years, six male), were recruited. A topical anesthetic (4% lidocaine or 0.5% tetracaine) was sprayed into the subject's right nostril while the left nostril served as a control. Unilateral nasal NO and nasal volume were measured before administration of the anesthetic and at 15 and 30 minutes after the administration. The mean (+/- SD) unilateral nasal NO output was 307 +/- 45.9 nL/minute from the right nostril (exposure side) before the topical application of lidocaine. At 30 minutes after topical application (n = 6), it was 295.5 +/- 41.5 in the right nostril and 297.5 +/- 39.8 in the left (control side). In the tetracaine group (n = 7), the mean (+/- SD) unilateral nasal NO output was 302 +/- 53.3 before the administration and 307 +/- 39.7 at 30 minutes after the administration in the right nostril. The mean NO output in the left nostril at 30 minutes after the administration was 297.7 +/- 40.75. In neither group was there any significant difference in nasal NO output between either the pre- and postlocal anesthetic application on the exposure side (Group 1, P = 0.76; group 2, P = 0.41) or the two nostrils after topical anesthesia application (group 1, P = 0.83; group 2, P = 0.62). Topical anesthesia with either lidocaine or tetracaine does not alter nasal NO output. NO measurement should not be affected in circumstances that require topical anesthesia of the nasal cavity. PMID- 10793917 TI - Association of bronchopneumonia with sinusitis due to Bordetella bronchiseptica in an experimental rabbit model. AB - An animal model for rhinogenic sinusitis was developed in rabbits naturally colonized with Bordetella bronchiseptica. It was found that ostial occlusion predisposes the sinus to invasion with this opportunistic bacterium and subsequent sinusitis as a result of reduced local host defense. In addition to the inflammatory lesions in the sinus, bronchitis and pneumonia were found in 84% of the experimental rabbits, suggesting that ostial dysfunction can also contribute to infectious disease of the lower respiratory tract. In such a model it is possible to study the significance of asymptomatic carriage of potential pathogens after ostial occlusion. PMID- 10793918 TI - Experimental myospherulosis of the paranasal sinuses: a histologic rabbit study. AB - Myospherulosis or lipogranuloma formation is frequently iatrogenic and is caused by a petrolatum, lanolin, or paraffin-based ointment becoming trapped within tissue. Four different ointments, including the newly available Bactroban nasal, were studied in a rabbit paranasal sinus model to evaluate their proclivity to induce myospherulosis. The maxillary sinuses of 16 New Zealand white rabbits were bilaterally inoculated with Bactroban, Bactroban nasal, tetracycline, or bacitracin ointments and compared to saline controls in two rabbits. Sinus specimens were harvested at 2 and 4-week intervals and processed for histologic study. Myospherulosis formation was uniformly induced with the Bactroban nasal, bacitracin, and tetracycline ointments in 8/8, 7/8, and 8/8 sinuses, respectively. In contrast, myospherulosis was not induced in the 5/8 of the sinuses using Bactroban. The data from this investigation indicate that Bactroban nasal (paraffin vehicle) is similar to bacitracin and tetracycline ointments (petrolatum and petrolatum-lanolin vehicles) in that they all can cause myospherulosis. In contrast, Bactroban (a water-soluble, polyethylene glycol base) causes myospherulosis to a much smaller extent. Our results emphasize the differences between the two types of Bactroban preparations and provide objective data that can be evaluated by otolaryngologists who apply these ointments following nasal cavity surgery. PMID- 10793919 TI - [The fight against pain in children: still precarious results]. PMID- 10793920 TI - [Outcome of children with chronic renal failure]. PMID- 10793921 TI - [Non-medical factors in perinatal health. A study of socioeconomic anc cultural features of women admitted to the maternity hospital in Lens (Pas-de-Calais)]. AB - BACKGROUND: The aim of this study was to determine the influence of medical and non-medical factors on perinatal health in an underpriviledged area, the ex-coal mines of Lens (Pas-de-Calais). POPULATION AND METHODS: This prospective study concerned 1,000 women who delivered in the maternity hospital of Lens between January 5, 1993 and May 13, 1993. The relationships between preterm delivery, low birth weight and maternal socioeconomic and cultural characteristics were analyzed. RESULTS: The women included presented adverse pregnancy outcomes. Preterm delivery, low birth weight and neonatal transfer rates were respectively 6.9%, 7.3% and 12.7% higher than the average regional rates. In contrast with these results, antenatal care could be considered correct or even better. The poor socioeconomic status of the population appeared to be a very significant perinatal risk factor. CONCLUSION: At present, in an unfavourable regional conjuncture, socioeconomic and cultural factors have adverse effects on perinatal morbidity and mortality independent of medical factors like prenatal care. The prevention of poor perinatal outcome should be based on these data. Not the access to the health care system but the manner of dispensing health care should require great consideration. PMID- 10793922 TI - [Neurodevelopmental outcome of premature infants born at less than 33 weeks of gestational age and not cerebral palsy at the age of 5 years]. AB - AIM: To appreciate the impact of prematurity, fetal hypotrophy and familial environment on the neurodevelopmental performances of very premature infants without cerebral palsy at the age of five years. POPULATION AND METHODS: We followed a regional cohort of 171 very premature infants (< or = 32 weeks of gestation) until they were five years of age. Cognitive functions were tested with the WPPSI test and the development quotient was assessed by the ability to draw a "bonhomme". Twenty-two premature infants suffered from cerebral palsy diagnosed before the age of two years. Another infant had a moderate diplegia at the five-year examination. We had no information for 16 prematures (9.3% of survivors). Twenty-eight premature infants were considered as having no severe disability on phone or mailed contact, and another child had a severe isolated mental retardation. We examined 104/148 infants, and 96/148 survivors without cerebral palsy passed the tests. The cognitive functions of these premature infants are compared to the performances of a control group made up of 108 children born at term > or = 37 weeks, matched for birthplace and single or twin characteristics of the pregnancy. RESULTS: The values of the different quotients are significantly decreased in the preterm group. The global IQ and the performance IQ are 0.8 SD, verbal IQ is 0.5 SD and the development quotient is 0.4 SD below the values observed in the control group. A performance IQ less than -2 SD for the mean of the control group is observed three times more than in the controls (13.5% vs 3.7%, P < 0.01). Multiple linear regression shows that prematurity explains, independent of hypotrophy and socioeconomic environment, 8% of the variation of the performance IQ (P < 0.01), 2% of the variation of the verbal IQ and 2% of the development quotient (P < 0.05). CONCLUSION: The five year neurologic outcome of the children born prematurely in this regional study is similar to the results observed in regional studies conducted in Europe: 13.4% of the survivors have cerebral palsy, and the cognitive functions of the children with no cerebral palsy are significantly lower than the term control group. Other risk factors such as hypotrophy, which modulates the developmental quotient, and the socioeconomic status, which modulates the verbal IQ, are underlined. PMID- 10793923 TI - [Liver transplantation in infants and children. Evaluation of the first 40 cases (March 1991-March 1997)]. AB - BACKGROUND: Liver transplantation (LT) is the treatment of end-stage liver disease in children. We report our experience with LT using grafts from living related (LRD) and cadaver donors (CD). POPULATION: From March 1991 to March 1997, 40 children and infants received a total of 42 liver grafts. A reduced-size liver was used in 28 cases. We studied pre-transplantation status, survival rate, and medical and surgical complications in these patients. RESULTS: The survival rate in our series was respectively 85 and 80% at 1 and 7 years after LT. Low weight infants required a prolonged ventilatory assistance. Five of the six deaths noticed during the first three months after LT occurred in children weighing less than 12 kg. One year after LT, no significant difference in the incidence of rejection was found, neither between low-weight children and the others, nor between patients transplanted from CD or LRD. Biliary tract stricture was the major surgical complication. CONCLUSION: This series consisted of a majority of low-weight children. The survival rate in the patients weighting less than 12 kg is lower than in the others. This may be explained by the nutritional status of these patients and early postsurgical complications. The use of grafts from living donors offers more flexibility since the operation is performed electively, but it did not seem to modify the incidence of acute rejections and surgical complications. PMID- 10793924 TI - [Incidence of inflammatory bowel disease in children in Brittany (1994-1997). Breton association of study and research on digestive system diseases (Abermad)]. AB - BACKGROUND: The aim of this work was to determine in Brittany the incidence and main clinical pattern of inflammatory bowel disease (IBD) occurring during childhood. These data are compared to the previous epidemiologic data available from the Northern France registry or around the world. METHODS: Private and public Brittany gastroenterologists (2,836,418 inhabitants including 618,049 children under 17 years of age) referred all patients consulting for inflammatory bowel disease from January 1994 to December 1997. An interviewer-practitioner completed at the gastroenterologist's office a standard questionnaire for each patient. Each case was independently reviewed by four experts in a blind manner and made a final diagnosis of Crohn's disease (CD), ulcerative colitis (UC), or ulcerative proctitis and acute colitis (onset of symptoms < 6 weeks) or unclassified chronic colitis. RESULTS: Among 1,309 cases recorded, 88 were under 17 years of age (6.7%): 43 (49%) had CD (including three possible cases), 14 (16%) had UC (including three proctitis), 24 (27%) acute colitis and 7 (8%) unclassified chronic colitis. The crude mean annual incidence (per 100,000 children) based on definite and probable cases only was 2.5 for IBD, 1.6 for CD and 0.57 for UC, without variation between 1994 and 1997. The male/female ratio was 2.3 for CD and 1.3 for UC. The mean time between onset of disease and diagnosis was equal to 7.2 and 8.6 months for CD and UC respectively (median: 3 and 5 months). A familial history of IBD was present in 5 cases (8%). In CD, the small and large bowel were involved in 58% of patients, whereas an isolated involvement of small or large bowel occurred in 15% and 23% of cases. Among the 14 UC, there were three proctitis and four pancolitis. Among 43 CD, a granuloma was present in 48% of cases. CONCLUSIONS: In Brittany the incidence of CD and UC in childhood was similar to the published data from Northern France. Clinical presentation and symptoms were not different. However, the rate of acute colitis was higher and the accurate incidence of IBD could be underestimated, requiring a follow-up to classify these cases. PMID- 10793925 TI - [Neonatal hemoperitoneum]. AB - PATIENTS AND RESULTS: A series of four cases observed in the department of pediatric surgery in Monastir between 1990 and 1998 are reported. The diagnosis was made in the two first days of life. The clinical features associated paleness, shock and abdominal distension. Ultrasonography revealed that the hemoperitoneum was associated with liver hematoma in three cases and with retroperitoneal hematoma in two cases. Three infants required emergency hemostatic laparotomy. The fourth was stabilized by initial resuscitation. One infant died at the age of six months following subarachnoid hemorrhage and subdural hematoma. No complication occurred in the three remaining patients. CONCLUSION: The pathogeny, clinical features and treatment of the hemoperitoneum in the newborn are discussed. The authors stress preventive measures, the only way to decrease the incidence of this neonatal complaint. PMID- 10793926 TI - [Acute recurrent pancreatitis and heterozygous mutation of the cystic fibrosis gene: a case report]. AB - Recurrent pancreatitis is seldom associated with CFTR (Cystic Fibrosis Transmembrane Regulator) gene mutation. CASE REPORT: A 17-year-old boy presented with isolated idiopathic pancreatitis. CFTR gene mutations study revealed delta F508 heterozygous mutation. CONCLUSION: Mutations for the CFTR gene should be explored in case of recurrent pancreatitis. PMID- 10793927 TI - [Mycoplasma pneumoniae pneumonia in a four-year-old child with transient abscess in the right lower lobe]. AB - The frequency of Mycoplasma pneumoniae infection among community-acquired pneumonia, underestimated for a long time, is now better known. Severe evolution is yet uncommon. Differential diagnosis with Streptococcus pneumoniae is often difficult. CASE REPORT: A 4-year-old child was admitted for a right lower lobe pneumonia, with very high values of white blood cell count and CRP, worsening despite a treatment with high doses of amoxicillin, then with cefotaxime and vancomycin. Diagnosis of M. pneumoniae infection was considered only on the tenth day after admission and confirmed on the thirteenth day. Clinical outcome rapidly improved with macrolide antibiotherapy. Radiologic outcome consisted, two months after the beginning of the pneumonia, in abscess of the right lower lobe, which recovered in one month with continuing oral antibiotherapy. CONCLUSION: Lung abscess is very rare in M. pneumoniae pneumonia, as only two other cases were described in the literature. In all three cases, macrolide therapy was delayed. Those cases highlight the importance of considering M. pneumoniae infection in a beta-lactams-resistant community-acquired pneumonia, whatever its severity may be, and to start macrolide antibiotherapy. Our case also shows the possibility of a conservative treatment in case of pulmonary abscess, if clinical tolerance is good. PMID- 10793929 TI - [Radiology case of the month. A case of congenital osteopetrosis]. PMID- 10793928 TI - [Nutritional management of cystic fibrosis in children]. AB - A major goal in the management of cystic fibrosis patients is to maintain a good nutritional status as it improves long-term survival. A link is clearly established between the degree of malnutrition and the severity of the disease. Clinical and biological follow-up, better knowledge of energy requirements, dietary counseling and nutritional intervention help to optimize the growth of these patients through childhood and adolescence. PMID- 10793930 TI - [Children and illness: psychological aspects of hospitalization]. AB - Hospitalization affects both body image and perception, as well as the social and family relationships of the child. The resulting psychological disturbances may influence the course of the disease and the efficiency of the care. A good quality of hospital reception and psychological supportive measures help to make acceptable the unacceptable notions of physical suffering and disease. PMID- 10793931 TI - [Tolerance of acellular pertussis vaccine booster injection in 15-20-month-old children]. AB - Severe reactions to whole cell pertussis vaccine are the main reason for the poor adhesion of parents and practitioners to pertussis vaccine booster injections in children, leading to a poor immunization in the population, particularly in young adults. The acellular pertussis vaccine appears to be better tolerated while keeping a good immunogenicity. In a randomized multicentric study on pertussis vaccine booster injection tolerance in 386 15-20 month-old children, we confirmed the better local and general tolerance of the acellular vaccine as compared with the whole cell vaccine. This should help to develop a positive opinion toward pertussis vaccine among parents and practitioners, and to improve the level of active pertussis immunization among children, adolescents, and young adults. PMID- 10793932 TI - [Erythrocyte transfusion in the newborn]. AB - The main aim of erythrocyte transfusion in the newborn infant is to improve oxygen transport to the tissues. However, clear evidence for its effectiveness in this objective is lacking. In addition, the potential complications (mainly viral infections) of transfusion have led to a limitation of its use. Indications and modes of application are presented based upon the recommendations of the French national agency for health practice assessment (ANAES). PMID- 10793933 TI - [Brain abscess isolated and verified by tuberculin reactions: diagnostic difficulties]. PMID- 10793934 TI - [Impact of an information campaign on the prescription of ambulatory oral rehydration solutions in infants with acute gastroenteritis]. PMID- 10793935 TI - [Home nutritional rehabilitation of severely malnourished children in Dakar suburbs]. PMID- 10793936 TI - [Neonatal Listeria infection is very rare in Morocco]. PMID- 10793937 TI - [Prevention of respiratory syncytial virus infection in a peripheral hospital: economic problems and ethical questions]. PMID- 10793938 TI - [Severe hypernatremia due to feeding error]. PMID- 10793939 TI - [Neonatal bacterial infections at Owenda Pediatric Hospital in Libreville]. PMID- 10793940 TI - [Respiratory distress in pediatric admission and emergency services. Epidemiology and evaluation criteria]. AB - The medical part of the activity in the emergency units increased and has been ascribed to the raised number of infants or children admitted for acute dyspnea. This review is based on the published reports and the experience from the paediatric emergency unit from the Rouen area with 450,000 inhabitants, in France. We put forward the known epidemiological data and discussed the available means for the practitioner, which could help him in the decision to hospitalize. This review suggests that algorithms of treatment and severity evaluations must be set up in paediatric emergency units in order to validate them and specify the children who requires hospitalisation. PMID- 10793941 TI - [Role of endoscopy in the diagnosis of respiratory distress]. AB - In acute respiratory distress, bronchial endoscopy is useful to determine the nature of dyspnea: inspiratory dyspnea in laryngeal abnormalities and obstructions; mixed inspiratory and expiratory dyspnea in extrinsic and intrinsic compressions of the tracheobronchial tree; overall, foreign bodies. Moreover, it allows the evaluation of the lesions in bronchial contusions, tracheobronchial burns and hemoptysis. Flexible (fiberoptic) and rigid endoscopy are complementary methods. Bronchoalveolar lavage is useful in the microbiological investigation of pneumonias developing in immuno compromised children. PMID- 10793942 TI - [Upper airway obstruction (foreign bodies excluded)]. AB - Upper airway obstruction is a frequent cause of admission to the emergency department and the intensive care unit. Symptoms are mainly represented by dyspnea and stridor. Severity must be rapidly assessed to allow adapted treatment and avoid cardiac arrest and hypoxic encephalopathy. The possible etiologies are numerous, with acquired and congenital ones, but the majority is represented by laryngitis, lymphoid hypertrophy and laryngotracheomalacia. In case of respiratory failure, treatment must first establish airway patency with bag and mask ventilation, and then intubation. If vital prognosis is not threatened, biologic, radiologic or endoscopic examination can be performed to identify the cause of the obstruction and treat it. PMID- 10793943 TI - [Acute bronchiolitis in infants]. AB - Bronchiolitis is the most common disease of the respiratory tract during the first year of life, and occurs in annual epidemics in winter. The etiology is viral, and respiratory syncytial virus (RSV) is the commonest agent. Respiratory symptoms remain generally mild, and treatment just supportive and at home. Certain infants are at high risk of severe illness (age less than 3 months, preterm birth, neonatal respiratory disease, bronchopulmonary dysplasia, underlying chronic diseases), and require hospitalisation. Most treatments are of unproved (corticosteroids), or limited benefit (inhaled bronchodilators, antibiotics). Chest physiotherapy is indicated in case of bronchial secretion. Vaccine are not now available, but prophylaxis with human RSV immunoglobulin or monoclonal antibodies has to be considered for children at increased risk for severe disease. PMID- 10793944 TI - [Acute asthmatic crisis in children]. AB - Acute asthma attack in children is an attack responsible for life-threatening acute respiratory distress with partial or no response to bronchodilator drugs. The severity of the episode needs to be quickly evaluated. This presupposes a perfect knowledge of the clinical signs of severity. Treatment is urgent and first based on the administration of high doses of inhaled short-acting beta 2 agonists. In the more obstructed children, anti-cholinergic drugs can be added to nebulized beta 2-agonists. Because of their delayed effect, systemic steroids require an early prescription. Symptomatic treatments are: urgent hospitalization, oxygen if needed, proper hydratation. Continuous nebulization or intravenous perfusion of beta 2-agonists are prescribed with cardiac monitoring when no objective improvement is noted. Admission into the pediatric intensive care unit when bronchial obstruction continues will permit the association of bronchodilator drugs and the proposal of mechanical ventilation if needed. When the episode is resolved, a prophylactic treatment using inhaled corticosteroids must be prescribed. Clinical and spirometric follow-up has to be organized, and the patient and his/her family have to be educated. PMID- 10793945 TI - [Pleurisy: diagnostic and therapeutic management]. AB - Diagnosis of pleural effusion is difficult in children. The etiologies are numerous; however, infectious agents are more frequent. Thoracocentesis proves to be the first-line diagnostic tool. Light's criteria are the best for distinguishing whether the effusion is a transudate or an exudate. If the patient has an exudative pleural effusion, other tests are indicated to determine the etiology and in some cases the treatment: macroscopic appearance, cytology and differential white cell count (level of glucose, lactate dehydrogenase, adenosine deaminase, pH, bacterial cultures). Others investigations--biopsy of pleura by thoracoscopy or video-assisted thoracoscopy, bronchofibroscopy, CT scan--are sometimes useful. Intrapleural instillation of urokinase appears to be useful and safe. Evaluation is necessary for video-assisted thoracoscopy used early. PMID- 10793946 TI - [Spontaneous pneumothorax in children]. AB - Spontaneous pneumothorax is rare in childhood. Before 12 years of age the main underlying pathologies are asthma, cystic malformations, post infectious bullae, and infectious pneumoniae. After 12 years of age it is mainly associated with cystic fibrosis and constitutional slim morphology. Symptoms vary according to the extent of lung collapse and the diagnosis is confirmed on chest X rays. In mildly symptomatic pneumothorax, spontaneous resolution is achieved within few days. When cardiorespiratory difficulties are present, mechanical evacuation of air from the pleural cavity is necessary through a tube drainage maintained until complete pulmonary reexpansion. Surgical treatment is indicated in case of persisting air leakage after one week of efficient drainage, large cystic malformation or post infectious bullae, recurring or bilateral pneumothorax. PMID- 10793947 TI - [Management of children in respiratory distress]. AB - Conditioning in respiratory distress in childhood requires a rapid evaluation of severity based on clinical examination in order to immediately start helpful care to improve respiratory status. Oximetry measurement is now integrated in this initial evaluation determining oxygen therapy. Etiologic orientation, quick and clinic, is precised by family questions. Adapted therapeutic will begin and we know that most frequently etiologies are: croup, bronchiolitis and asthma attack. PMID- 10793948 TI - [Late revealing of congenital diaphragmatic hernia: diagnostic difficulties]. AB - Two patients, aged four and six months, with congenital diaphragmatic hernias, and who presented outside of the neonatal period, are reported. Clinical presentation was a progressive respiratory failure, and radiographic findings showed a consolidation of the left lung, associated with a pneumothorax in one and an ascites in the other. One had previous normal chest X-rays. All cases with herniated bowel showed connecting bowel segments passing through the diaphragmatic defect. All were repaired via an abdominal approach. Delay in presentation of congenital diaphragmatic hernias is not uncommon. Long-term prognosis is favourable and postoperative morbidity is minimal, despite late presentation and undesirable time delay between first sign and surgery. PMID- 10793950 TI - [Tracheobronchial foreign bodies]. AB - Forty cases of children with an inhaled foreign body (FB) are reviewed over a three-year period. Clinical data, radiologic findings and complications are detailed. The nature and size of FBs are also reported. Diagnosis and management are discussed according to the most recent studies in the literature. Diagnostic flexible bronchoscopy is a useful first step when the diagnosis is unclear (i.e., choking history, unexplained respiratory symptoms), but FB removal is usually not possible during this procedure. Extraction is performed via the rigid bronchoscope under general anesthesia. However, FB could be removed with the flexible bronchoscope in five children in our study. Diagnosis and removal of an inhaled FB are required as quickly as possible in order to prevent respiratory sequelae (bronchiectasis). Prevention is based upon information to be given to families, but to the medical community as well, which often minimizes the seriousness of inhalation hazards. PMID- 10793949 TI - [Role of inhalation therapy in respiratory emergencies in children]. AB - Three modes of inhaled therapy are available for the administration of bronchodilators in acute respiratory diseases in children: spacer devices, nebulizers, inhaled powders. Principles, medications available in France, and dosages, are presented. Acute asthma is the main indication. PMID- 10793951 TI - [Management of hemoptysis in children]. AB - Hemoptysis in children are infrequent and often self-limiting. They are a manifestation of the broader spectrum of pulmonary haemorrhage. Diffuse pulmonary haemorrhages are often associated with diseases of other organs (cardiopathies, systemic diseases). Focal haemorrhages have multiple aetiologies, dominated by bronchopulmonary infections and cystic fibrosis. Fiberoptic bronchoscopy allows one to localise the bleeding, look for local causes and diagnose pulmonary hemosiderosis by BAL. For local lesions and if the medical management fails, bronchial arteriography is indicated to perform the embolisation of the bleeding vessels. PMID- 10793952 TI - [Thoracic injuries in children]. AB - Chest trauma in children is rare but shows that trauma is severe and the mortality rate is high (30%). Multidisciplinary management of children includes an initial evaluation of respiratory distress, freeing the airways, placing an intercostal tube, stabilizing the chest wall, and analgesia. When vital signs are stable, secondary evaluation includes an etiologic, radiologic and biologic check up, ending with the therapeutic strategy. Thoracotomy is rarely required, and for most children, only monitoring will be necessary, though this is important because of the risk of secondary decompensation and late diagnosis of potentially fatal lesions. PMID- 10793953 TI - [Decompensation in chronic respiratory insufficiency in children]. AB - In patients with acute exacerbations of chronic obstructive or restrictive pulmonary disease, noninvasive ventilation can be used in an attempt to avoid endotracheal intubation and complications associated with mechanical ventilation. The main obstructive pathology concerned is bronchopulmonary dysplasia: bronchial hyperreactivity is a main feature of the situation, leading eventually to acute or prolonged assisted ventilation. Usually performed by tracheostomy, ventilation can possibly be managed through a nasal mask. The use of noninvasive ventilation is also indicated when symptoms of hypoventilation and daytime hypercarbia develop in a variety of neuromuscular disorders. PMID- 10793954 TI - [Pulmonary complications in immunosuppressed children]. AB - Pulmonary complications, which are dominated by opportunistic infections, can be first manifestations of inherited or acquired pediatric immune deficiencies. Prompt diagnosis is essential. The epidemiology and natural history of these complications have changed as a result of major advances in prevention, diagnosis and treatment. PMID- 10793955 TI - [Respiratory distress and drepanocytosis]. AB - Acute chest syndrome (ACS) is characterized by chest pain with dyspnea and recent radiologic abnormalities, and is an acute lung complication whose problem is one of etiology. Alveolar hypoventilation linked to infarcts of the thoracic ribs, thoracoabdominal trauma, subdiaphragmatic pain, the administration of analgesics causing respiratory depression, or sleep disturbance, is a frequent cause of ACS. Bronchoalveolar lavage has revealed the frequency of fat embolism following infarcts in the long bones. Pulmonary vascular occlusion, due to thrombosis or emboli, is rare, as are the infectious pneumonia and pulmonary edema. The pathogenetic mechanisms consist of an alteration of the rheological properties of the blood, the existence of an hypercoagulability state, specific interactions between the abnormal sickle cells and the vascular endothelium, and a dysregulation of the vascular reactivity. Research centered around NO biology has led to an expanded understanding of the critical interdependence of NO, hemoglobin, and the microvasculature. An anemic patient with ACS suffers from loss of pulmonary scavenging and hypoxic pulmonary vasoconstriction and loss of peripheral NO delivery. Interruption of this cycle by transfusing normal (hemoglobin A-containing) erythrocytes might improve all the abnormalities. PMID- 10793957 TI - Results of a meta-analysis of cost-benefit research: is this a question worth asking? AB - We attempted a meta-analysis of telemedicine research studies of the costs associated with telemedicine. First, we performed a search of six well known databases with a variety of relevant keywords. After discarding non-English publications, books and duplicate publications resulting from the same study, we were left with 551 articles for analysis. Our second step was to separate the articles into two groups: those with and those without quantitative cost data. Only 38 articles contained any type of real data. Because many of these 38 studies proved to be inadequately designed or conducted, we were unable to perform a traditional meta-analysis. Furthermore, there were a number of disturbing features common to these studies, including the omission of the number of consultations or patients, almost non-existent longitudinal data collection and lack of uniformity in cost analyses. We conclude that it is premature for any statements to be made, either positive or negative, regarding the cost effectiveness of telemedicine in general. PMID- 10793956 TI - A randomized controlled trial to assess the clinical effectiveness of both realtime and store-and-forward teledermatology compared with conventional care. AB - The clinical effectiveness of realtime teledermatology, store-and-forward teledermatology and conventional outpatient dermatological care were evaluated in a randomized control trial. A total of 204 patients took part--102 patients were randomized to the realtime teledermatology consultation, 96 of whose cases were also referred using a store-and-forward technique, and 102 to the conventional outpatient consultation. There were no differences in the reported clinical outcomes of realtime teledermatology and conventional dermatology. Of those randomized to the realtime teledermatology consultation, 46% required at least one subsequent hospital appointment compared with 45% of those randomized to the conventional outpatient consultation. In contrast, the dermatologist requested a subsequent hospital appointment for 69% of those seen by store-and-forward teledermatology. An analysis of costs showed that realtime teledermatology was clinically feasible but more expensive than conventional care, while the store and-forward teledermatology consultation was less expensive but its clinical usefulness was limited. Sensitivity analysis indicated that realtime teledermatology was as economical as conventional care when less artificial assumptions were made about equipment utilization, costs and travel distances to hospital. PMID- 10793958 TI - Supervising trauma life support by telemedicine. AB - The resuscitation room in a community hospital was linked with a main hospital accident and emergency department using telemedicine equipment working at 384 kbit/s. Fifteen simulated casualties replicated the 'moulage' scenarios in the Advanced Trauma Life Support Course Manual of the American College of Surgeons. Each of the 15 scenarios was broken down into three main parts: the primary survey, resuscitation and the secondary survey. While a physician in the community hospital undertook each task, a senior doctor in the accident and emergency department recorded his degree of confidence in the supervision of the task on a five-point scale. There were features of the management which the supervisor found difficult, mainly related to the camera view and the use of a proxy examiner. However, supervising major trauma management by telemedicine was feasible. The average scores were mostly above 3 and often above 4 in the assessment of the primary survey and the resuscitation. The average scores were mostly above 3 for the secondary survey but were less often above 4 than for the primary survey and the resuscitation. Trials of remote trauma management with real patients appear to be justified. PMID- 10793959 TI - How do teleconsultations for remote trauma management change over a period of time? AB - We obtained follow-up information about the new patients seen at a minor accident and treatment service (MATS) staffed by emergency nurse practitioners (ENPs). A previous study, of 150 teleconsultations in a six-month period starting in April 1996, was used for comparison. In the present series, 150 teleconsultations occurring in a four-month period starting in April 1999 were studied; the patients constituted 5.6% of the 2658 new attenders or 3.8% of the 3936 total attenders. In comparison with the study three years before, falling teleconsultation rates were partly offset by increasing numbers of attenders and an extension of the ENPs' roles and skills. Teleconsultation rates rose when the number of consultant-run clinics was curtailed and ENP-run clinics replaced some of them. Eligibility to request and report more radiographs reduced the need for teleconsultations, and subsequently teleconsultations for help with interpretation of radiographs fell as the ENPs became more experienced. Specialty residents trusted ENP judgement and accepted telephoned direct admission of cases to their wards. Fewer teleconsultations were required for soft-tissue injuries. Telemedicine is an excellent educational tool. PMID- 10793960 TI - Increasing the cost-effectiveness of telemedicine by embracing e-health. AB - In 1999 a national study of telemedicine in Australia led to the promotion of the concept of 'e-health', the health sector's equivalent of 'e-commerce'. A new study explored the view that, with the convergence of technologies and the consequent increase in ability to perform multiple functions with those technologies, it is unwise to promote telemedicine in isolation from other uses of technologies in health-care. The major sources of information for the study were the presentations and discussions at five national workshops held to discuss the findings of the original report on telemedicine. Nineteen case studies were identified. The case studies showed that with the convergence of technologies telehealth is becoming part of e-health. The cost-effectiveness of both telehealth and telemedicine improves considerably when they are part of an integrated use of telecommunications and information technology in the health sector. PMID- 10793961 TI - Telepsychiatry evaluation in the north-west of England: preliminary results of a qualitative study. AB - A telepsychiatry referral service for patients suffering from anxiety and depression was evaluated from a user perspective. Low-cost video-phones linked a psychiatrist to two general practices in the north-west of England. Quantitative data were collected using a semistructured interview schedule. Twenty-two patients and 13 doctors were interviewed after a video-link consultation. Some users were very positive about the service and recognized its potential benefits, while others were more ambivalent. Patients saw the service as a means of obtaining additional 'expert' advice. General practitioners felt that the service might adversely affect the doctor-patient relationship in psychiatry. Both patients and clinicians recognized that the video-link modified normal interaction. Users need to adapt to this form of communication. An induction session is recommended for both patients and clinicians. PMID- 10793962 TI - The cost-effectiveness of teleradiology at the Women's and Children's Hospital in Adelaide. AB - The cost-effectiveness of teleradiology was examined in a trial conducted at the Women's and Children's Hospital (WCH) in Adelaide, from February 1998 to February 1999. The trial showed that, with the large distances between remote hospitals and metropolitan hospitals, teleradiology, relative to the transfer of remote patients, could be highly cost-effective, but that a new form of cost justification is required for teleradiology for tertiary hospitals providing second opinions in special cases. A series of compelling case studies proved the economic and social value of teleradiology at the WCH. PMID- 10793963 TI - Telemedicine for intensive support of psychiatric inpatients admitted to local hospitals. AB - A service was developed to treat acute psychiatric inpatients in their local hospitals using telemedicine. This reduced the need for these patients to be transferred to a psychiatric facility in Adelaide. An evaluation of outcomes showed that it was possible to manage acute psychiatric patients in this manner, reducing costs of transport. In addition, patients were treated close to their homes. Patients rated their satisfaction with the service and the use of telemedicine very highly. PMID- 10793964 TI - Training for surgeons using digital satellite television and videoconferencing. AB - The TETRASUR project (TELematics TRAining for SURgeons) has investigated the effectiveness of distance-learning technologies in the delivery of training for doctors who are studying for membership of the Royal College of Surgeons. Digital satellite television receivers and ISDN videoconferencing equipment have been installed in hospitals to deliver the course modules, including a series of live television programmes transmitted by satellite. ISDN videoconferencing was integrated, live, into the satellite broadcasts to bring in guest lecturers and for interactive discussions with the trainee doctors. Videoconferencing was also used for seminars and discussion groups. These methods proved to be effective and popular with the doctors, although there was some dissatisfaction with the visual quality of the ISDN videoconferencing at 128 kbit/s. Efforts are now being made to improve the quality of the video feed from remote sites using ISDN at 384 kbit/s. PMID- 10793965 TI - The AIDMAN project--a telemedicine approach to cardiology investigation, referral and outpatient care. AB - The AIDMAN pilot project will connect health clinics on remote Greek islands with a mainland hospital. We have developed a virtual consultation workstation for the project, together with a satellite communication network. A UK pilot site has been used to test the concepts and applications between a surgery in Chorleywood and St Mary's Hospital in London. PMID- 10793966 TI - Could a low-cost audio-visual link be useful in rheumatology? AB - We have investigated prospectively the diagnostic accuracy, specialist satisfaction and patient-specialist rapport of a low-cost audio-visual link between a junior doctor with a patient and a consultant rheumatologist. Using a telephone link and subsequently a video-phone link, 20 patients, with various rheumatological problems, were presented by a junior doctor to the consultant rheumatologist for provisional diagnosis. All patients were then seen face to face by the consultant, when a final diagnosis was made. An independent consultant rheumatologist made a 'gold standard' diagnosis. Thirty-five per cent of diagnoses were made correctly over the telephone and 40% over the video-phone- there was no significant difference in the diagnostic accuracy between these two methods of communication. Rapport over the video-phone was universally poor. Where it was important, clinical signs could not be visualized over the video phone and in more than 85% of cases small joint swellings could not be seen clearly. PMID- 10793967 TI - A review of telemedicine cost-effectiveness studies. AB - As interest in telemedicine grows, many of its proponents and vendors increasingly suggest that it is now time to move to full-scale implementation of telemedicine services in a variety of contexts throughout the world, and question the need for further evidence of its utility and cost-effectiveness. We have reviewed the published literature relating to cost-effectiveness studies in telemedicine and have identified some important weaknesses. Ten recommendations regarding the design of economic evaluations of telemedicine are suggested. PMID- 10793968 TI - Latitude and noise comparisons between digital cameras and a radiographic film scanner. AB - The test image generated by a radiographic laser printer was used as a test object to measure the latitude and image noise of two digital cameras and a dedicated radiographic film scanner. The film scanner had the widest latitude and least image noise. A Kodak digital camera had a wider latitude but more image noise than an Olympus camera. A blue filter on the Olympus camera increased the latitude without significant increase in image noise. PMID- 10793969 TI - Telemedicine use in two nurse-led minor injuries units. AB - Telemedicine links were established between two nurse-led minor injuries units (MIUs) located in community hospitals and an accident and emergency department in a district general hospital. Telemedicine consultations were logged. During the first year of operation, telemedicine use was surprisingly constant, there being an average of 36 calls a month from one MIU and 16 from the other. Possible reasons for this include the clinical protocols under which nurses could refer patients to hospital. The average duration of teleconsultations was 3.5 min. Although the MIUs dealt with roughly similar numbers of patients, approximately 600-700 per month, the proportion for whom telemedicine was used was very different, at 5.2% and 2.3%. PMID- 10793970 TI - The cost-effectiveness of teleneurology consultations for patients admitted to hospitals without neurologists on site. 1: A retrospective comparison of the case mix and management at two rural hospitals. AB - We are currently evaluating the use of telemedicine for improving the care of patients admitted with neurological symptoms to hospitals that do not have specialist neurologists on site. To do this we have been comparing the outcome of patients admitted to two small hospitals. In one hospital all patients with neurological symptoms are seen by a neurologist at a distance using an interactive video-link transmitting at 384 kbit/s; in the other patients with neurological problems are managed as per usual practices. For the results of this study to be valid, it is essential that the case-mix and process of management for neurological patients are similar at the two hospitals. We therefore compared the case-mix, process of management, and outcome for all patients admitted over a four-month period to either hospital who had been coded using ICD-10 as having a final diagnosis of a neurological condition. No appreciable differences were noted between the two hospitals for measures of case-mix or outcome. Likewise, most measures of process were similar, although there was a significant difference for the overall length of hospital episode between the two hospitals. When patients with prolonged hospital episodes were excluded, or only patients with a diagnosis of headache, epilepsy or transient ischaemic attack were considered (who as a group made up the bulk of neurological admissions), the difference in the length of hospital episode was not significant. It should therefore be possible for us to estimate the effect of telemedicine on the management of patients with neurological problems. PMID- 10793972 TI - Using telemedicine to facilitate training in cardiotocography (CTG) interpretation. AB - Electronic fetal monitoring is a controversial practice in modern obstetric care and is frequently an aspect of medicolegal cases involving the management of labour and delivery. The interpretation of the cardiotocograph (CTG) produced by such monitors is a skill required by those caring for the pregnant woman. Studies have shown that most 'experts' do not interpret CTGs in a consistent manner, when compared with either other experts or themselves. However, it has also been shown that consistency can be improved with training. Telemedicine has been used to advantage in the training of obstetrics and gynaecology registrars in CTG interpretation. PMID- 10793971 TI - Interactive television for an urban adult mental health service: the Guy's Psychiatric Intensive Care Unit Telepsychiatry Project. AB - We carried out a feasibility study of an interactive television (IATV) system to enhance the provision of psychiatric intensive care services to a remote adult acute psychiatric ward in the same National Health Service mental health trust. The system used videoconferencing equipment connected by ISDN at 128 kbit/s. The system was used for patient referral, assessment and monitoring by staff at the remote site 8 km away. PMID- 10793973 TI - Enhancement of an antenatal diagnosis and counselling service (ADACS) through the ready availability of telemedicine services. AB - The early diagnosis of fetal abnormalities and their consequent management comprise a relatively new specialty, which has developed to a large extent from the advances in and availability of ultrasound imaging techniques. The Women's and Children's antenatal diagnosis and counselling service (ADACS) is composed of a multidisciplinary group of health professionals and deals with issues relating to pre-pregnancy counselling, fetal diagnosis of abnormalities and the management of these conditions, including all aspects of pregnancy loss. Detailed minutes are recorded for each case and key information is stored in a relational database. Weekly meetings are held to discuss selected cases and, where possible, are presented to the group by the referring practitioner, either in person or using the telemedicine facilities. Telemedicine has provided a significant enhancement to the ADACS service. PMID- 10793974 TI - Using the law to enhance provision of telemedicine. AB - Reference is often made to the law as a barrier to the use and development of telemedicine. In New South Wales, however, it has been recognized that some of the principles behind these 'legal impediments' can be used to make services better and more cost-effective. The law offers a range of useful tools relating to the emergence of these new techniques. The responsibility of those in a position to regulate is, quite simply, to pick the right tool. PMID- 10793976 TI - Working with telemedicine: user characteristics and attitudes. AB - Combined qualitative and quantitative interviews were conducted with 30 persons involved in four telemedicine specialties. The results showed little technology related anxiety and all viewed the technology as useful. There was a positive general attitude to telemedical work, with a perception of the possibility of job satisfaction being improved and the technology being found to be easy to use. Negative general attitudes to telemedical work were not found. The implementation of telemedicine may therefore have a positive effect within organizations. Such findings may be important for the future widespread adoption of telemedicine. PMID- 10793975 TI - An Internet-based telediagnosis system for Chinese medicine. AB - The Internet is becoming an increasingly popular medium for communication and is being used more and more for telemedicine. Telediagnosis is one of the most developed components of telemedicine. We have designed and partly implemented a telediagnostic system, using the Internet as the means of communication, to enhance access to specialists and health providers in Chinese medicine. PMID- 10793977 TI - Cost-effectiveness of home telemedical cardiotocography compared with traditional outpatient monitoring. AB - We compared the cost of passive sensor telemedical non-stress cardiotocography performed at home and the same test performed by traditional equipment in an outpatient clinic in the Budapest area. The costs were calculated using two years' registered budget data from the home monitoring service in Budapest and the outpatient clinic of the department of obstetrics and gynaecology at the Haynal Imre University of Health Sciences. The traditional test at the university outpatient clinic cost 3652 forint for the health-care and 1000 forint in additional expenses for the patient (travel and time off work). This means that the total cost for each test in the clinic was 4652 forint. The cost of home telemedical cardiotocography was 1500 forint per test, but each test took 2.1 times as long. For a more realistic comparison between the two methods, we adjusted the cost to take account of the extra length of time that home monitoring required. The adjusted cost for home care was 3150 forint, some 32% lower than in the clinic. Passive sensor telemedical non-stress cardiotocography at home was therefore less expensive than the same test performed in the traditional way in an outpatient clinic. PMID- 10793978 TI - Design and initial implementation of a regional tele-oncology project. AB - Two tele-oncology projects have been in progress since 1997 in the Province of Trento in north-east Italy. The common aim of the projects concerns the design and the implementation of a non-surgical tele-oncology system intended to provide a flexible computing environment for the joint management of oncology patients in a wide-area network. The two projects involve both hospital specialists and general practitioners treating oncological patients. The first phase of the project involves the design and implementation of the oncology teleconsultation service. PMID- 10793979 TI - An assessment of the potential effect of a teledermatology system. AB - The potential effect of a commercial teledermatology system was assessed. The system allowed general practitioners to send dermatologists a set of digital images, accompanied by a short patient history. Patients were seen, in the normal way, by consultant dermatologists. The system was then used to capture a set of images. These were viewed by two dermatologists, 13 months later. Reasonably high levels of agreement were found between the dermatologist seeing the patient and the dermatologist using the telemedicine system (77%). The two dermatologists were also asked to indicate whether, had the system been in use, the patient would have been seen urgently or routinely, or whether the general practitioner would have been advised that an outpatient appointment was not required. The results showed that fewer patients would have been called for urgent appointments (32% compared with 64%) and that 31% of cases could have been managed by the general practitioner. Assuming that the introduction of the system would have had no effect on the overall number of referrals, nor on the number of follow-up appointments, these figures suggest that the total number of appointments could be cut by 13%. It took approximately an hour to view 20 cases and it would be necessary to devote one consultant session a week to viewing images. This system would therefore not lead to significant savings, nor reduce the waiting list for outpatient appointments. The idea would have potential if the review of images could be made as easy as the triage of referral letters. PMID- 10793980 TI - The potential for telemedicine and telecare in Deptford. AB - We carried out a study to explore the potential of telemedicine and telecare to improve health-care for the population served by a health centre based in Deptford in south-east London. A wide range of potentially useful telemedicine and telecare applications was identified, nine of which could provide some immediate benefit. Other requiring new approaches to the delivery of care involved some reengineering of services. Some suggestions focused on the need to integrate information technology with service provision. Other suggestions raised strategic questions, the most important of which concerned the routes of access to health-care. PMID- 10793981 TI - Improving quality of life for dialysis patients through telecare. AB - Home dialysis can improve the care and quality of life for patients with renal failure. We have explored the possibility of extending home care to more patients needing continuous ambulatory peritoneal dialysis (CAPD) using telemedicine. We tested videoconferencing support for five CAPD patients using low-cost ISDN equipment (128 kbit/s). Initial results indicated that it was possible to integrate videocommunication into the daily routine of the clinic and the response from patients was surprisingly positive. Selection of appropriate, affordable technology and the ISDN service support by the telecommunications provider proved to be considerably more difficult than anticipated. The first indications also suggest medical advantages for home teledialysis. PMID- 10793982 TI - Postoperative evaluation of patients following ophthalmic surgery. AB - We have used telemedicine to support the postoperative evaluation of patients who have had ophthalmic surgery. Mobile telemedicine workstations connected using three ISDN lines have enabled us to review patients at an outreach clinic on the first postoperative day. Video slit-lamp images of the patient are captured by a trained ophthalmic nurse at the outreach clinic and viewed by surgeons at Moorfields Eye Hospital in central London during live teleconsultations. During the study period, over 80 teleconsultations were carried out on postoperative cataract, trabeculectomy and combined procedures. Preliminary results are very encouraging. Although 8 (33%) of 24 patients were anxious about being involved in the teleconsultation, 20 (83%) had confidence in the system and only one (4%) found the experience unacceptable. PMID- 10793983 TI - Application of business case analysis in planning a province-wide telehealth network in Alberta. AB - A strategy for implementing telemedicine throughout Alberta was developed. The model was based on a comprehensive evaluation of the four clinical specialties chosen as representative telemedicine services--radiology, psychiatry, emergency services and continuing education. The goals of the telemedicine network were to improve access to health services, provide support for rural health-care providers and increase the efficiency of specialized services. The findings showed that the success factors in a national telemedicine programme depend on a clear organizational structure, with appropriate technical standards and support. PMID- 10793984 TI - Evaluating telemedicine for minor injuries units. AB - Low-cost telemedicine equipment consisting of ISDN videoconferencing units and a store-and-forward system was installed in two minor injuries units (MIUs) and a hospital accident and emergency department in Lincolnshire. Over six months, 45 patients were treated using telemedicine in one MIU and 26 in another. Anecdotally, there were no reported radiograph discrepancies or missed diagnoses. Data collected during teleconsultations by both referring and consulting clinicians suggested that in some cases teleconsultation had helped to avoid transfer or onward referral. There were some changes in diagnosis and treatment after using telemedicine, indicating some decision-making value for the remote practitioners. In the context of minor injuries telemedicine, videoconferencing in realtime may prove to be more valuable than store-and-forward interactions. A cost-benefit analysis is being conducted using a pragmatic prospective case control study. PMID- 10793985 TI - Telemedicine services to a county jail. AB - Local and county jails rarely offer telepsychiatry services to their inmates. We have established a telepsychiatry pilot project between the Kansas University Medical Center and the Lyon County Jail in Emporia, Kansas. A total of 264 telepsychiatry consultations were conducted with jail inmates. Of these, 70 were initial evaluations and 194 were follow-up visits; only one inmate refused to be seen. Approximately one-third of all inmates were seen for psychiatric consultation within one week of their incarceration and 68% were seen within one month of incarceration. Among lessons learned during the first year of service were: the monthly demand for consultations was five times greater than projected; moderately to severely ill inmates with a broad range of psychiatric illness can be seen and treated effectively using videoconferencing; and the technology was accepted by the jail personnel and the inmates alike and integrated into the jail's routine in terms of the delivery of psychiatric care. PMID- 10793986 TI - Tele-ophthalmology screening for retinal and anterior segment diseases. AB - A digital indirect ophthalmoscope (DIO) was developed and tested for use in tele ophthalmology screening for posterior and anterior segment diseases. Using custom software, images from the DIO were digitized, compressed, stored and transmitted to a centralized eye clinic for interpretation. A total of 43 subjects were primarily screened for glaucoma using the DIO, a hand-held fundus camera and a stereo fundus camera. The photographic slides from the stereo fundus camera used as gold standards. Images (390 x 300 pixels x 3 bits) were stored using a laptop computer together with patient information. Image quality received at the central eye clinic was good and showed adequate diagnostic information. An ophthalmologist estimated cup-disc ratios and graded the quality of the images. The sensitivity and specificity of each instrument was calculated. A high sensitivity and specificity was found when using the DIO, indicating that it could be used in tele-ophthalmic screenings. Further modifications are needed to make the instrument more user-friendly and to enable it to be used with undilated pupils, so that it can be easily operated by health-care personnel in remote areas. The camera can also be used to image gross external eye pathology. PMID- 10793987 TI - Patient satisfaction with a home televisiting service based on interactive television over a cable network. AB - Experience shows that high-quality audiovisual contact between remote health carers and patients facilitates a telemedicine service. However, the lack of broadband communication to the home usually prevents domestic televisiting. Deployment of cable networks in Spain has allowed the implementation of a home televisiting service designed for patients with chronic diseases. In a trial, 15 patients received televisits by three specialists and three nurses from the Severo Ochoa Hospital in Madrid. Five patients suffered from chronic pain, five were from the nephrology unit and five had been treated at the intensive-care unit after acute myocardial infarction. Each patient participated in three televisiting sessions, two provided by a specialist and the other by a nurse. The average length of a televisit was 12 min (range 5-21 min). The patients expressed their satisfaction with the service. PMID- 10793988 TI - Transforming local care. AB - The prevalence of diabetes mellitus is steadily increasing in Western society, with obvious implications for costs to health-care providers. A project plan to transform local care has been established in Wales to enable diabetic patients to self-manage their disease by improving education and access to information and tests locally. The plan involves the establishment of local diabetes centres providing educational resources to the patients, carers and health professionals. It envisages that the local diabetes community centres will provide all aspects of diabetic care, from diagnosis to continuing management, with the aid of modern communications technology. PMID- 10793989 TI - Contracting with health-care customers and specialists for the provision of telemedicine services across European borders: the TEN-Telemed Legal Project. AB - Two generic contract models have been developed to resolve the legal and ethical issues that arise when contracting for telemedicine services across Europe. Model 1 relates to the risks and responsibilities of the expert providing and specialist opinion and the telemedicine service organization, while model 2 outlines the risks and responsibilities from the perspective of the client seeking the advice and (again) the telemedicine service organization. These contracts express the legal rights and responsibilities of each of the parties involved. PMID- 10793990 TI - The intellectual property cookbook: a guide for the novice health-care telemedicine provider working with industry. AB - Telemedicine is a new field and many health-care providers are developing their own products with the help of industry. Most practitioners are novices in the legal tools necessary to protect their own work with regard to any future commercialization. To summarize these issues for the telemedicine practitioner, a review of intellectual property protection has been performed. Intellectual property can be protected by tools such as copyrights, patents, non-disclosure and confidentiality agreements, integrated circuit topographies and industrial design. Knowledge of the intellectual property background should allow telemedicine providers to protect their own work when working with industry. PMID- 10793991 TI - Five years' experience with the Telemedicine Information Exchange. AB - The Telemedicine Information Exchange (TIE) has provided comprehensive telemedicine information on the World Wide Web (Web) since early 1995. It received major funding from the National Library of Medicine in 1997. Among other things, the TIE contains six major databases: literature citations; active telemedicine programmes; a 'what's new in telemedicine' column; funding opportunities; forthcoming conferences; and a list of vendors of telemedicine equipment and services. Recent additions include a document delivery service, inaugurated in early 1999. More than 1000 other Web sites link to the TIE, and we have 5000 visitors per month from several countries. Given its relative longevity on the Web, TIE researchers have been in a unique position to observe trends in telemedicine. PMID- 10793992 TI - Putting the 'tele' into health-care effectively. AB - Although the introduction of new technologies has been successful and become accepted practice in many areas of industry, traditional methods have tended to prevail in health-care. Telemedicine has been adopted by enthusiasts who recognize the potential benefits of a 'global health service'. However, the more widespread introduction of telemedicine requires considerable organizational change in the way health-care is delivered. More evaluation is required of clinical outcomes, organizational effects, benefits to health-care providers and users, and quality assurance. PMID- 10793993 TI - The need to ensure that the globalization of information and telematics does not destabilize health-care worldwide. AB - Modern telecommunication transcends institutional, organizational and cultural boundaries. In the context of health-care, this means that information, enquiries, demands and service provision can be routed anywhere, free of institutional or natural control. The Internet makes information available to anyone regardless of its quality, source or intention. Organizationally initiated applications such as telemedicine are usually intended to reinforce local systems, but their very design and intention indicate a potential rapidly to transcend inherited but electronically ineffectual boundaries. The consequences of such uncontrolled globalization of health-care activities will range from beneficial empowerment and quality improvement, to detrimental effects such as overloading of experts, and undermining of stable health-care systems. The major unplanned societal re-engineering effects in a paper-based culture are likely to be significant and global institutions need to respond by creating positive global frameworks and policies. PMID- 10793994 TI - A pilot study of radiotelemetry for continuous cardiopulmonary monitoring of patients at home. AB - We have conducted a trial of a wireless device for continuous cardiopulmonary monitoring. Its performance, user acceptance and safety were assessed for monitoring in the patient's home. The study included 20 patients: six with chronic obstructive pulmonary disease, six with chronic heart failure, seven with atrial fibrillation and palpitations, and one with a snoring problem. The system recorded the heart rate and respiratory rate, blood pressure, electrocardiogram and body temperature. The results were transmitted automatically to a central monitoring station. The accuracy of the measurements was checked by a comparison system and also by conventional measurements performed by a nurse. The system was acceptable to patients and functioned satisfactorily in the home. An important facet of home telemonitoring may turn out to be its greater reliability in collecting objective data. PMID- 10793995 TI - Diagnosis of glaucoma using telemedicine--the effect of compression on the evaluation of optic nerve head cup-disc ratio. AB - A photograph of the optic nerve head requires a lot of disk space (over 1 MByte) for storage and may require substantial bandwidth and time for transmission to a remote practitioner for a second opinion. To test whether compression degrades the image quality of the images, 302 slides were digitized at an optical resolution of 2400 pixels/inch (945 pixels/cm) and 30 bit/pixel. The images were saved both in non-compressed TIFF format and in compressed JPEG (compression ratio of 60) format. A blinded observer measured the optic nerve head cup-disc ratio for all three groups: the original slides, uncompressed TIFF and compressed JPEG images. The results showed that digital images were less accurate than slides. However, compression, even up to a ratio of 40, did not make matters worse. PMID- 10793996 TI - Telemedicine in the control of intra-ocular pressure. AB - Glaucoma is one of the most common causes of blindness in the Western world and a major risk factor is increased intra-ocular pressure. We therefore used telemedicine in its control. Patients measured their intra-ocular pressure several times a day with a portable instrument and the values were then entered into a portable digital assistant. These data were transmitted by a modem to a central server. If the intra-ocular pressure was pathologically high, an email message was automatically sent to the ophthalmologist. The pressure curve, including a statistical analysis, was displayed in an easily readable chart format. Ten patients with glaucoma participated in a trial. Self-tonometry with telemedicine enabled continuous evaluation of the patient by the ophthalmologist. This approach offered the advantage of controlling the treatment remotely. Advantages for the patient were that the measurements were easily done at home under normal conditions, and the patient could control when the measurement and data transmission would be performed. Telemedicine is a cost-effective technique enabling the early diagnosis of pathologically increased intra-ocular pressure. PMID- 10793997 TI - School-based telemedicine: teachers', nurses' and administrators' perceptions. AB - In 1997, a telemedicine project began in Kansas which brought health-care directly into elementary schools using interactive video-links. The project initially involved four schools, but was expanded to cover 10 schools, including two middle and one high school. We examined the organizational issues related to telemedicine links to schools. Specific attention was paid to the perceptions of the nurses, teachers and key administrators. The research involved analysing archive data and interviewing participants in the project from the schools and the medical centre. The results showed the difficulties in delivering health care, especially by telemedicine, to under-served urban children. However, the data also revealed that these can be overcome. Once they had experienced it at first hand, almost all concerned began to see telemedicine as an effective and important asset to the delivery of health-care. PMID- 10793998 TI - What do we really know about the cost-effectiveness of telemedicine? AB - Telemedicine is still in its infancy, but undergoing rapid development. It is very difficult to evaluate telemedicine. We performed a literature survey (Medline). During the period 1990-8, over 1500 articles on telemedicine were published. Of these, 246 mentioned economic aspects in the abstract (16%). We selected 29 studies although few had demonstrated cost-effectiveness. Benefits for the patients in the form of reduced travel and waiting time must often be weighed against increased provider costs. Up to now, telemedicine in general has not had any significant effect on medical practice, or the structure and organization of health-care. In order to utilize the potential of telemedicine, its integration with traditional health-care is very important. There are country specific variations in the health systems that make it difficult to generalize the results from one country to another. PMID- 10794000 TI - Legal issues relating to telemedicine in Canada's publicly funded health-care system. AB - The advent of telemedicine poses new legal challenges for Canada's health-care system. Although telemedicine increases access, it does not fit easily into Canada's traditional one-tier approach to health-care. Cross-border telemedicine services may inadvertently contribute to a two-tier system. The current legal system will have to be adapted to accommodate changes instigated by cross-border telemedicine. PMID- 10793999 TI - Partnering through telemedicine to improve the quality of health-care. AB - In order to assist health-care reform in eastern Slovakia, a partnership was established with health-care institutions in Rhode Island. The aim of the partnership was to improve maternal and child health in Slovakia. Thirty-seven telemedicine sessions were conducted via a satellite link. A number of workshops, case studies, demonstrations and interactive discussions took place between various health-care professionals. The use of telemedicine accelerated the adoption of new procedures in Slovakia. PMID- 10794001 TI - The current status of medicolegal issues surrounding telepathology and telecytology in Japan. AB - The medicolegal issues surrounding telepathology and telecytology in Japan have been partly resolved since the government declared that telediagnosis in these fields using a microscope is essentially a medical practice between doctors. Thus it does not contravene article 20 of the law pertaining to medical practice in Japan, which states that there must be an actual face-to-face encounter between the doctor performing any diagnostic or treatment episode and the patient. It is, however, only beginning to be recognized that the medicolegal issues surrounding telepathology and telecytology depend on factors such as the type of system used (e.g. active versus passive). In an active diagnostic system, the telepathologist or telecytopathologist can control a robotic microscope at the remote site, while in a passive diagnostic system the telepathologist or telecytopathologist makes the diagnosis based on the microscope images that have been selected, and transmitted, by someone in the remote hospital. In the former case the interpreting telepathologist or telecytopathologist must assume most of the responsibility for the diagnostic process, while in the latter more responsibility must be assumed by those at the remote site. The duties and responsibilities of all participants in the telediagnostic process, whatever the system employed, must be determined. We have established working guidelines for telepathological and telecytological diagnoses to help achieve this and hence ensure safe and effective clinical practice. PMID- 10794002 TI - Multidimensional telecare strategies for rural residents with brain injury. AB - Rural residents with brain injury have difficulty in accessing care from qualified psychologists for consequent cognitive, emotional and behavioural symptoms. We examined high-quality videoconferencing to enhance care for persons with brain injury in three areas: cognitive assessment, psychotherapy and rural mental health training. The assessment study evaluated 52 outpatients seen for diagnostic visits over videoconferencing, and compared their experiences with those of 52 age- and diagnosis-matched controls seen in person. Persons seen via telemedicine were more likely than controls to want to repeat their experience and more satisfied than were the neuropsychologists who examined them. In the psychotherapy study, neurorehabilitation patients were seen via videoconferencing for therapy related to brain injury or stroke. Persons receiving psychotherapy were less likely than persons receiving assessment services to want to repeat their experience. In the training study, 39 rural mental health providers were trained via videoconferencing, and trainees demonstrated significant improvement on tests of knowledge about brain injury. Trainees formed a network of mental health provider referrals for persons with brain injury in a wide geographic area. Given adequate training and ongoing support, rural clinicians can treat many brain-injury adjustment issues locally, reserving specialist consultation for emergency or complex problems. PMID- 10794003 TI - The design of a practical and reliable fall detector for community and institutional telecare. AB - Falls are one of the greatest obstacles to independent living for frail and elderly people. Their early detection is an important step in providing people with the reassurance and confidence necessary to maintain an active lifestyle. We have investigated a worn fall detector linked to a community alarm system. A worn device is the only one which is satisfactory, provided that it generates few false alarms. The fall detector we have developed is the size of a small radio pager. It uses a two-stage detection process which senses shock and the orientation of the wearer. A fall is detected within 20 s and triggers a radio signal to a community alarm system. Tests were devised using a jointed mannequin to simulate five modes of falling to understand the effects of impact at different parts of the body. This allowed us to select the appropriate trigger threshold and wearing positions for the sensor. Prototypes were evaluated with 20 people to observe false alarms. The final design allowed reliable detection in 180 different falling scenarios. PMID- 10794004 TI - A community-based approach to evaluation of health outcomes and costs for telepsychiatry in a rural population: preliminary results. AB - A pilot trial was established to support visiting psychiatric services and local public and private practitioners through the use of videoconferencing. The purpose of the trial was to determine whether people in the community received better health-care with telemedicine. A community-based approach was used to evaluate health outcomes, costs, utilization, accessibility, quality and needs for such services in a rural community in Queensland. Over a two-year period data were collected from 124 subjects who met the criteria of having a mental health problem or mental disorder. Nine further subjects refused to participate in the study. Only 32 subjects used videoconferencing to receive mental health services. Preliminary results did not show any significant improvements in wellbeing or quality of life, although the time span was relatively short. However, the results confirmed that the people were no worse off from a consumer or a practitioner perspective from using videoconferencing. Most consumers found that videoconferencing with a psychiatrist moderately or greatly helped them in managing their treatment, with 98% of them preferring to be offered videoconferencing in combination with local services. Overall, videoconferencing is a crucial part of enhancing psychiatry services in rural areas. However, it is not necessarily cost-effective for all consumers, general practitioners, psychiatrists, or the public mental health service. PMID- 10794005 TI - Telemedicine in Greenland--from the early beginnings to the making of a business plan. AB - Greenland has a geography and demography that make telemedicine a logical choice when planning a future public health service. Telemedicine was introduced in 1994. Although everybody considered it to be a good idea, it took five years to move from initial enthusiasm to politicians and officials committing the necessary resources. Over such a long period, there is a risk that good ideas will become neglected, although, where resources are limited, it is useful as a time in which to select the best of the new ideas. We have certainly gained a great deal of experience which we will use if and when, over the next two years, we have to argue for an extension of our business plan. PMID- 10794006 TI - Security aspects of teleradiology between the university centre and outlying hospitals in Tyrol. AB - We assessed a data security system using biometric fingerprint techniques and smartcards to control access to a teleradiology system. Clinical cases were initially discussed between the referring physician and radiologist in Tyrol using a video-phone. Subsequent correspondence, including the transmission of images between the referring physician and the radiologist, was carried out by e mail using the security system to prevent unauthorized access to patient information. Seventy-eight teleradiology sessions were conducted using this data security system. Speed and stability of data exchange were unaffected by the additional security feature. The average log-on time to the system was 7.8 s. The average training time on how to use the system was 15 min. The radiologist was able to issue a final patient report using the system within 1.5 of the initial contact. The data security feature was user-friendly and did not hinder the normal teleradiology consultation. PMID- 10794007 TI - Telecardiology: results and perspectives of an operative experience. AB - A telecardiology system has been established between six Italian hospitals. Four of them are connected using three ISDN lines; the remaining two are connected with only a single ISDN line. The telecardiology system was evaluated between two hospitals. Radiographs showing the heart movements of 10 patients were digitized and forwarded to the specialist hospital for expert opinion. The transfer time of a single patient sequence ranged from 6 h 40 min to 22 h 15 min using a single ISDN line at 128 kbit/s. The transfer time decreased considerably using three ISDN lines. Following data transfer, a joint consultation was organized between the cardiologists in realtime. Realtime telecardiology using three ISDN lines was clinically viable and more efficient than the traditional method of delivering this type of specialist care. PMID- 10794008 TI - A cost-benefit analysis of the effect of shipboard telemedicine in a selected oceanic region. AB - Data from a selected oceanic region, the UK search and rescue region, were used to establish the average annual of ship diversions and emergency service call outs arising from urgent medical problems of passengers or crew. During the period 1997-8 there were 228 medical evacuations. An attempt was made to estimate the extent to which some or all of the diversions and call-outs could have been averted if telemedical facilities had been available on-board the ships. The analysis showed that telemedicine would be an expensive alternative to existing evacuation methods, but did not allow for the fact that helicopter and lifeboat evacuations cannot be carried out in bad weather or at distances over 200 nautical miles (370 km) from land. Taking into consideration the cost of ship diversions in such circumstances produces completely different results. Telemedicine could clearly provide substantial cost-savings for the shipping industry, and a separate analysis focused on the shipowner alone would make an overwhelming economic argument for investment in a telemedicine service simply on the strength of diversion avoidance. PMID- 10794010 TI - Global medicine and licensing. AB - Telemedicine embraces the concept of global health-care. In order to realize the internationalization of the practice of medicine, licensing issues need to be addressed. Most countries have their own regulations on licensing to protect the patient and ensure high-quality care. International licensing regulations are likely to result in the standardization of medical practice. PMID- 10794009 TI - The economics of telepathology--a case study. AB - There are several obstacles that slow down the diffusion of telepathology. One is related to uncertainty about the economic consequences of its adoption, possibly more so than in other fields of telemedicine. We have evaluated the economics of telepathology when used to provide a frozen-section service to a mountain hospital, in comparison with three current alternatives. In the specific situations studied, no one model was always less expensive than the others. In particular, owing to the very low cost of the ambulance service provided by the Red Cross, the ambulance model was least expensive when dealing with up to 73 frozen sections a year, while at higher case-loads telepathology was cheaper. If ambulance transfer is neglected, telepathology appears to be the most convenient approach to the remote frozen-section service. Although the consultant pathologist costs more than telemedicine, during free time he/she could perform other (routine) work, thus reducing the real cost of frozen sections. PMID- 10794011 TI - Telemedicine and patients' rights. AB - Telemedicine improves access to both health-care and second opinions. However, the potential risks of telemedicine, such as breaches in patient confidentiality and security, need to be addressed in order to ensure that patients' rights are not violated. Used appropriately, telemedicine has the potential to enhance the rights of patients to high-quality, effective health-care. PMID- 10794012 TI - An occupational therapy Internet school. AB - Four centres in the UK, Belgium, the Netherlands and Sweden have collaborated to provide Internet-based courses for student and professional occupational therapists. The basis of the occupational Therapy Internet School (OTIS) is the concept of the 'virtual college'. This comprises the design and implementation of a sophisticated Internet-based system through which courses can be managed, prepared and delivered on line, and through which students can communicate both with the staff and their peers. The aim is to support and facilitate the whole range of educational activities within a remote electronic environment. PMID- 10794013 TI - A comparison of telemedicine with face-to-face consultations for trauma management. AB - We compared the accuracy of teleconsultations for minor injuries with face-to face consultations. Two hundred patients were studied. Colour change, swelling, decreased movement, tenderness, instability, radiological examination, severity of illness, treatment and diagnosis were recorded for both telemedicine and face to-face consultations. Colour change showed an accuracy of 97%, presence of swelling or deformity of 98%, diminution of joint movement of 95%, presence of tenderness of 97%, weight bearing and gait of 99%, and radiological diagnosis of 98%. The severity of illness or injury was overestimated in one case and underestimated in five cases. Treatment was over-prescribed in one case and under prescribed in three cases. The final diagnosis was correct in all but the two cases in which mistakes were made in the teleradiology. Overall, there was good accuracy using teleconsultations. PMID- 10794014 TI - Transfer of telemedical support to Cornwall from a national telemedicine network during a solar eclipse. AB - During late 1998 and early 1999, planning officers in Cornwall predicted a huge increase in summer visitors to the county to observe the August solar eclipse. There was the possibility that a mass gathering in Cornwall could overload existing arrangements for handling accident and emergency patients. We therefore set up a telemedicine system to support the county's minor injury units (MIUs) from hospitals throughout the UK. Six main hospital accident and emergency departments outside Cornwall with existing links to their own MIUs were twinned with 10 of the 11 MIUs in Cornwall before the expected date of the gathering. The network was live for nine days, starting four days before the eclipse, and 2045 patients were seen in the 10 MIUs. There were 93 telemedicine calls from the 10 MIUs, involving 91 patients. Overall, 4.6% of the patients required a telemedicine consultation. Fifty-seven calls were made during working hours. Thirty-four patients were referred for further management, of whom 18 were referred on the same day. The transfer of telemedical support to a national network was successful. PMID- 10794015 TI - A pilot study of an educational service for rural mental health practitioners in South Australia using telemedicine. AB - Studies have found that rural patients want to be treated in their local hospitals. However, there have been difficulties with recruiting and retaining professionals in rural areas of Australia. Isolation, lack of professional development programmes, lack of academic programmes, lack of peer consultation and the need to travel long distances are some of the reasons identified in the National Rural Health strategy. A pilot service of educational programmes delivered by telemedicine was initiated and then evaluated. Forty-six community mental health workers from nine rural areas and 20 general practitioners from five rural areas participated. High satisfaction with the use of telemedicine was recorded by both groups. Telemedicine reduced the disincentives to rural practice, and also improved the professionals' competence and confidence in managing patients with psychiatric illness. PMID- 10794016 TI - Smart care technologies: meeting whose needs? AB - Recent funding programmes supporting research and development in telecare have argued for a shift in perspective from a technology-driven approach to one that is needs-led. While this is in the interests of both users and technologists, achieving this goal is not straightforward. This paper outlines some of the conceptual, methodological and practical problems that potentially constrain a needs-led approach and illustrates the emergent issues with a case study of the development of an intelligent home monitoring system to support the independent living of older people. The research indicates clear differences between users and technologists in the way problems, needs and requirements are understood and defined. This in turn has consequences for the way assistive technologies are developed and implemented. PMID- 10794018 TI - Values and technology assessment in psychiatry. AB - Economic resources for health care are limited and they should be distributed as fairly and effectively as possible. But the basis for such a distribution is far from clear. What interests are involved? What kind of provision of care is most efficient? How far should the distribution be left to market solutions? What is the proper role of health care politicians and authorities? In technology assessment a promising combination of value premises and empirical knowledge is used in answering such questions. The aim of this introductory article is to explore issues related to values and value conflicts that have implications for technology assessment. Ethical principles are discussed and related to problematic issues in mental health care such as the absence of psychiatry, the abuse of psychiatry, the definition of mental illness, diagnostic activities, treatment decisions, priority setting as well as research and development. PMID- 10794017 TI - A cost measurement study for a home-based telehospice service. AB - A telehospice service is one in which telemedicine is used to provide hospice care in the home. To date, there have been few studies addressing the cost of home-based telemedical care, and none that specifically addresses cost effectiveness for telehospice recipients. We measured costs for traditional hospice care as well as those associated with launching and operating a telehospice service. The costs were tallied over two separate three-month periods. For the first study period, costs were measured for traditional hospice home visits. During the second, expenses were monitored for traditional (in person) and telehospice visits. For traditional care, the cost per visit was $126 and $141, for the first and secod time periods, respectively. The average telehospice visit cost was $29. PMID- 10794019 TI - Political abuse of psychiatry. AB - The abuse of psychiatry in Nazi Germany 60 years ago was the abuse of the 'duty to care'. Its scale was enormous; 300,000 people were sterilized and 100,000 killed in Germany alone and many thousands further afield, mainly in eastern Europe. This episode occurred in a country with a high reputation for its medicine, including psychiatry, and for its interest in the ethics of medical research. The economic conditions which preceded the violent political upheaval had led to increasing concern about 'the burden on the State' of the mentally ill and disabled. These preoccupations are still with us today. There may still be lessons to be learnt from the Nazi episode. PMID- 10794020 TI - The Declaration of Hawaii and Clarence Blomquist. AB - The international code of ethics of psychiatry, the Declaration of Hawaii was in the main the achievement of Clarence Blomquist. There were several prerequisites for the success of this work. 1. The unique profile of the education of Clarence Blomquist, combining training to be a specialist in psychiatry with a doctor's degree in practical philosophy. 2. An outstanding competence in analyzing complicated issues and in putting thoughts into words. 3. The courage to challenge the Hippocratic ethics and adapt the principles of ethics to modern health care. 4. A scholarship at the Institute of Society, Ethics and the Life Sciences, Hastings-on-Hudson, New York, where he could test his ideas in an intellectual interdisciplinary atmosphere. 5. Support from the late Professor Leo Eitinger, Norway and Professor Gerdt Wretmark, Sweden, who together with Clarence Blomquist constituted a task force on ethics of the World Psychiatric Association. 6. A continuous backing-up by Dr Denis Leigh, the then secretary general of the World Psychiatric Association. Denis Leigh was convinced that a code of ethics was the only means to reconcile the various member countries on issues of misuse of psychiatry and, in addition, would raise the quality of psychiatric care throughout the world. PMID- 10794021 TI - From the Hawaii Declaration to the Declaration of Madrid. AB - The Declaration of Hawaii, adopted by the World Psychiatric Association (WPA) in 1977, was a significant event. But the needs and new ethical dilemmas of the 1990s led WPA to develop new recommendations on the duties of psychiatrists resulting in the Declaration of Madrid, adopted by WPA in 1996. It outlines the framework of ethical conduct of psychiatrists, formulates seven general guidelines with an increased emphasis on research and resource allocation, and gives five specific guidelines on euthanasia, torture, the death penalty, selection of sex, and organ transplantation. PMID- 10794022 TI - Human rights law in the field of mental health: a critical review. AB - The 'centrepiece' of international human rights law in the field of mental health is often said to be the United Nations Principles for the Protection of Persons with Mental Illness of 1991. Some observers appreciate the symbolic importance of these principles in providing visibility to the needs of the mentally ill, in stressing the right of access to adequate mental health care and in establishing the principle equivalence between psychiatry and the rest of medicine. However, the Principles appear basically flawed in several respects: (1) they do no have the status of a formal international treaty; (2) States are not required to adopt the Principles as 'minimum standards' for the protection of mentally ill persons; (3) in some respects, notably on the issue of consent to treatment, the Principles remove patients' rights rather than reinforce them; (4) the Principles do not provide for redress nor for any form of monitoring, inspection or supervision by an independent international body. Thus, it appears that even at an international level the deep-seated societal ambivalence towards the mentally ill has taken root and that so called 'human rights' principles have little material effect on the lives of psychiatric patients and create double standards in the exercise of choice. PMID- 10794023 TI - The right to accept and the right to refuse. AB - The main purpose of this paper is to show that the civil commitment acts of the Nordic countries are in conflict with the ideal of symmetry, i.e. the idea that if a person has the competence (and therefore the right) to accept hospitalisation he or she should also be considered competent (and therefore have the right) to refuse hospitalisation. First, we distinguish between one narrow and one wide concept of coercion in relation to hospital admission; second, we demonstrate that the narrow concept of coercion is used or presupposed with reference to hospitalisation in the Nordic countries; and third, we discuss, from a normative point of view, the rather disturbing asymmetrical conception of patient's competence and rights implied by this narrow concept of coercion. Though the ideal of symmetry supports the use of the wide concept of coercion in civil commitment acts, it does not decide the issue. To justify a change from a narrow to a wide concept would also require empirical data about the practical implications. PMID- 10794024 TI - Ethical issues in psychiatric care: geneticisation and community care. AB - This paper will examine two different though related themes in current debates about ethical issues in psychiatric care. There is, first, the general question of who should be the main focus of the debate: the individual, the family, the local community, or the wider society? Secondly, the current controversies about the genetic basis of mental disorders will be explored with reference to their implications for both images and understanding of mental disorders and for psychiatric care. Would the understanding of the genetic causes of mental disorder lead to better treatments and better acceptance or to the potential for increased discrimination and stigmatisation? PMID- 10794025 TI - Rights and responsibilities of the psychiatric profession. AB - Psychiatry as a medical discipline relies on the authority of medicine that is associated with the help to a suffering and deserving individual. If this source of authority is obscured, the discipline will be blamed for serving as a social tool for controlling undesirable phenomena and practices. Psychiatry as a medical science accumulates knowledge on the relationship of biology and psychopathology. It can provide an explanation of the extent to which mental illness participates in socially undesirable behaviour and phenomena. But it cannot explain undesirable social phenomena as a mental illness of sorts, let alone offer an effective treatment for them. We should carefully guard the boundaries of psychiatry to prevent its abuse in the future. PMID- 10794026 TI - The ethical base of mental health service research. Recent developments in mental health service research in the UK. AB - After the developments in mental health services in the UK in the last 40 years there has been an increasing focus on identifying and targeting the needs of the severely mentally ill (SMI). Political concern about the possible risks of community care have resulted in an emphasis on administrative and legal means of ensuring continuity of care and close clinical supervision of this SMI group. A number of experimental community-oriented mental health services have been shown to be cost-effective compared to hospital-based treatment. The further development of comprehensive community mental health services will depend on demonstrating whether this can be generalised to routine clinical settings. In this context we shall present the design of the PRiSM study, a prospective controlled trial of the cost-effectiveness of community mental health teams for the SMI in South London. PMID- 10794027 TI - Psychiatric ethics and health services research. Concepts and research strategies. AB - In a conceptual frame of reference a distinction is made between different aspects of coercion (formal, perceived and evaluated) and different levels of coercion (hospitalization, treatment and other measures). This frame of reference is used to analyse empirical data from interviews of representative samples of 100 formally committed, the 'probands', and 99 voluntarily admitted sex- and age matched 'controls'. The results demonstrate that there are many different aspects of coercion; thus, discussions about psychiatric coercion should not be restricted to formally registered commitment. The data also indicate the need to survey the different perceptions of the persons involved in the process of care, in order to understand whether and how coercion has been applied. The results also exemplify the need to include, in all assessments of health care, questions of respect for patient autonomy and integrity. PMID- 10794028 TI - Implementing the lessons of mental health service demonstrations: human rights issues. AB - Is the failure to implement the lessons of mental health service demonstrations an ethical problem? This paper reviews the experience with evaluating multi-site service demonstration programs in the United States. Some of the lessons have been adopted and others have not. There appears to be a pattern to the adoption of lessons: ideological and inexpensive changes in organization or treatment approach are more likely to be adapted than costly changes in the availability of services or 'transfer' resources (such as disability benefits or rental subsidies) to support independent housing. The paper reviews this experience for demonstrations such as the Community Mental Health Centers program, the Community Support Program, and the Robert Wood Johnson Foundation Program on Chronic Mental Illness, and it discusses prospects for current federal demonstrations. PMID- 10794029 TI - Psychiatric care and communities. AB - Psychiatrists have been concerned with communities both as settings for the lives and treatment of their patients, and as human organizations which have characteristics that can be studied, understood, and modified by the concepts and strategies developed in more traditional psychiatric work. Our growing understanding of psychiatric disorders has increasingly emphasized their chronic course with long periods of premorbid predisposition, inter-episode vulnerability and post-episode sequelae, and the role of the community as a social context with stressors, protectors, and critical variables which often dwarf the power of our therapeutic interventions. PMID- 10794030 TI - Findings from London's Mental Health': a service in crisis. AB - The King's Fund published its second London Commission in July 1997, and had produced a report 'London's Mental Health' in preparation for this, consisting of the most complete account ever published of the present state of the mental health services. They are found to be in a state of crisis, unable to cope with the very high levels of demand seen in the deprived inner city areas. There are now too few beds in the National Health Service to deal with the number of patients who need to be admitted, and too few residential places in the community to take those ready for discharge. The report considers the reasons for the high demand, and finds London to contain the six most socially deprived districts in the country, the highest unemployment rates, and double the number of persons living in single person accommodation. Emergency care is from the Accident and Emergency Departments, and the complete range of community care is not available anywhere. Remedies for these problems include adjustments to the resource allocation formulae, removal of the difficulties preventing new capital building in the public sector, and removing the divide between health and social services. PMID- 10794031 TI - Abuse of psychiatry. AB - The Geneva Declaration of 1948 states that doctors shall not permit considerations of religion, nationality, race, party politics or social standing to intervene between their duty as a doctor and their patients. They shall not use their medical knowledge contrary to the laws of humanity. The UN Principles of Medical Ethics also reminds doctors that their professional relation to persons deprived of their liberty is solely to evaluate, protect or improve their health. Similarly, the Hawaii Declaration from 1983 and the Madrid Declaration from 1996 outline the ethical duties of the psychiatric profession. Thus, there is no doubt as to the professional role as it is delineated in the medical ethics codes. The presence of such codes is, however, only one of the necessary steps to prevent the abuse of psychiatry; others include educational interventions or the establishment of international networks. PMID- 10794032 TI - On the justification for civil commitment. AB - This paper explores some of the controversies in the debate regarding the justification of civil commitment. The sometimes conflicting values reflected in the mental health legislation, human rights principles, moral philosophy and psychiatric professional standards are discussed. In spite of the often substantial use of civil commitment in many countries, there are almost no scientifically sound studies addressing the outcome of coercive treatment. The paper establishes that the traditional arguments in favour of civil commitment, like lack of insight and competence as well as the effectiveness of civil commitment, are poorly founded. The paper concludes that there seems to be a general agreement that civil commitment of patients who are dangerous to themselves or others should be the responsibility of the mental health care system, while civil commitment for treatment purposes is more controversial and hard to justify. PMID- 10794033 TI - Surviving psychiatry in an era of 'popular punitiveness'. AB - In an era when sentencing of mentally disordered offenders has been progressively influenced by protective considerations, the role psychiatrists play in the sentencing process is problematic. Where an offender's legitimate expectation of proportionality in sentencing can be trumped by psychiatric assessments, not of therapeutic need, but of either predictions of risk or untreatability or both, leading to disproportionate and potentially damaging custodial sentences, then psychiatrists should recognize that they are ethically compromised. PMID- 10794034 TI - Forensic psychiatry, ethics and protective sentencing: what are the limits of psychiatric participation in the criminal justice process? AB - As clinicians, psychiatrists are unequivocally dedicated to relieving the suffering of those who are afflicted with mental disorders. However, the public and those individuals, who are assessed, find it difficult to draw a distinction between forensic psychiatrists acting in a clinical role and the very same professionals acting in an evaluative role, on behalf of the state. This paper examines the ethical issues raised by psychiatric involvement in the sentencing process. It rejects the view that a forensic psychiatrist, who undertakes an evaluation for the state, is to be considered as an advocate of justice who is not bound by conventional ethical duties to the individual whom he or she assesses. It contends that the forensic psychiatrist has an important role to play in presenting evidence that may result in the mitigation of the sentence that may be imposed on a person who is mentally disordered. The paper will focus on these issues in the particular context of the situation in England and Wales, Canada and the United States. PMID- 10794035 TI - Rights of persons with mental illness in Central America. AB - This paper reports a diagnostic inquiry on the human rights situation of persons with mental disorders in four Central American countries. The study was conducted by teams constituted of mental health workers and human rights lawyers, using a standard questionnaire prepared by the World Health Organization (WHO) and adapted for regional purposes by the America-based WHO office. The inquiry identified violations in different areas, e.g., involuntary admissions, informed consent and others. In general, violations were not associated with the level of social development of the country but with the affiliation of the care-giving system (private vs. public). This study will be followed by activities aimed at improving the current situation. PMID- 10794036 TI - At the margins of human rights and psychiatric care in North America. AB - The roots and expanse of the rights of psychiatric patients in North America are broad and diverse. This paper focuses on four rights that are pushing at the contemporary margins of patients' rights. First, the right to treatment, a moral position casting about for legal grounding. Second, the rights of psychiatrically hospitalized patients, articulated in statutes, court decisions, organizational standards and patients' bills of right. Third, patient participation in treatment planning, a process involving both rights and responsibilities. Fourth, the right to involuntary outpatient treatment, a process sometimes viewed as a deprivation of and other times as an expansion of rights for patients. These rights are addressed in the context of the question, are we going in the proper direction? PMID- 10794037 TI - Human rights and psychiatric care in Africa with particular reference to the Ethiopian situation. AB - Around 700 million people are estimated to live in the continent of Africa. The majority live far from health facilities and are short of basic supplies. Most African people believe that diseases in general, and mental illness in particular, are afflictions caused by supernatural evil forces. Traditional methods are preferred sources of help for health problems by most people in the continent. Modern psychiatric services are far from adequate. The available asylums are located in the capital cities and very few patients have access to them. There is no mental health legislation in some African countries. In Ethiopia, where the population is 55 million, there is only one mental hospital; and a total of 390 beds for psychiatric inpatients. There are 11 psychiatrists in the country. In the regions of the country, mental health services are provided by psychiatric nurses. Patients usually come to medical services having tried the available local means. Psychotic patients almost always are forced to come to the mental hospital by their families, friends, neighbours, work-mates (and very seldom by the police). Consent is not usually required to initiate treatment or admit such patients. Alleged offenders, who come to the hospital for assessment, stay in the same ward as other patients. Armed prison guards assigned to watch the prisoners also stay in the same room with the prisoners. Care providing procedures in Ethiopia do not seem to be in accordance with the declarations of human rights. However, in a country where the economy cannot provide its citizens with basic needs for survival, it is unlikely that the standard of mental health care will change much in the foreseeable future. PMID- 10794038 TI - Human rights in psychiatric care: an Asian perspective. AB - The foundations of the modern concepts of universal human rights have evolved out of centuries of economic, political and ideological conflicts in the west. There are variations in implementing human rights issues in Asian countries. However, increasing awareness about these issues is evident in the establishment of National Human Rights Commissions in India and Thailand and the greater concern shown by psychiatric care institutions in this region on human rights issues. PMID- 10794039 TI - Pax Iliev: law, life and care in transition. AB - The presentation proceeds from a clinical case of domestic violence, its management in terms of professional roles and institutional involvement and the analysis of the dynamic mental processes. It is argued that regimes of total control abolish legal systems and leave a void behind. In the course of transition to civil society, rule by intimidation and threat at a local community level is the rule. Suffering and trauma are intense and health systems necessarily become involved. A pattern of decontextualizing depression may be common and revealing of institutional defence mechanisms at work. Continuous affirmation of procedures of good practice, e.g. supervision, gives a chance for the preservation of institutional sanity and professional ethics. PMID- 10794040 TI - Reform in psychiatry in post-Soviet countries. AB - All the former Soviet countries are making efforts to reform psychiatry and adopt generally acknowledged international standards. Key issues of reform are changing the role of professional associations and the development of psychiatric legislation. This process is illustrated by the activities of psychiatric associations in Ukraine, Lithuania and Russia. PMID- 10794041 TI - Endometrial histology in long-term users of the once-a-month injectable contraceptive Cyclofem. AB - This study evaluated endometrial histology in women using the once-a-month injectable contraceptive Cyclofem for one year or more. The study received IRB approval. Seventeen Cyclofem users accepted to be submitted to an endometrial biopsy. All the samples were collected with an endometrial suction curette (Z Sampler, ZSI Gynecological Product, Chasworth, CA, USA) in an outpatient clinic, 27-33 days after the last injection. The material was fixed immediately in Bouin solution prior to wax embedding and screened as a routine histological examination by a senior pathologist. The pathologist did not know the bleeding status of each woman or the number of injections each woman had received before the biopsy. All women recorded bleeding and spotting for the last 60 days prior to the biopsy. The mean age of volunteers was 25.9 years (range 21-32) and the mean number of injections received was 24.8 (range 14-47). The results were: 4 out of 17 biopsies were found to be inadequate for diagnosis because they consisted of only blood and mucus. Two of these women were bleeding regularly and the other 2 had amenorrhea. Eight endometrial samples presented a proliferative pattern and, in this group, 3 women had amenorrhea and 5 were bleeding regularly. The other 5 biopsies were reported as secretory endometrium, and 4 of them showed pseudodecidual reaction compatible with the administration of progestin. All of these women were bleeding regularly. The results of the biopsies were not related to the number of injections received nor to the age of the women. In conclusion, long-term administration of the injectable contraceptive Cyclofem did not produce adverse alterations in the endometrium as hyperplasia. PMID- 10794042 TI - Safety of intrauterine device insertion by trained nurse-midwives in the Sudan. AB - Insertion of intrauterine devices (IUDs) by trained health workers other than physicians is increasing, particularly in developing countries. Twenty nurse midwives in government service in the Sudan, called health visitors (HVs), were trained to provide intrauterine contraceptives in a research project designed to evaluate the safety of insertion of IUDs by medical personnel who are not physicians. After training, they inserted 763 IUDs. Independent evaluation of 520 clients was conducted by gynecologists who found that only six devices (1.2%) had been incorrectly inserted. Outcomes for clients of the health visitors, with respect to perforations, infections, expulsions, and pregnancies, compared well with those of eight physicians who participated in the study. The research strongly supports the concept of nurse-midwife training for IUD insertion. This would greatly expand the availability of family planning services and would conserve physician time and skills for problem cases. PMID- 10794043 TI - Reversible antispermatogenic effect of gossypol in langur monkeys (Presbytis entellus entellus). AB - The present investigation reports the antispermatogenic effect of the orally active highly purified gossypol acetic acid at 7.5 mg and 10 mg/day for 180 days in langur monkeys. The results revealed a dose-dependent response in semen analysis as well as testicular morphology. Uniform severe oligospermia was observed in the lower dose (7.5 mg) group, while azoospermia was observed in 2 out of 5 animals in the higher dose (10 mg) group and the remaining animals showed severe oligospermia. Scanning electron microscopy of spermatozoa revealed deleterious abnormalities in the head and midpiece. Testicular morphology revealed a decrease in the seminiferous tubule diameter and arrest of spermatogenesis. The lower dose group had a germ cell population up to primary spermatocytes while the higher dose group had only Sertoli cells and spermatogonia. Withdrawal of treatment for 180 days led to the recovery of all the parameters studied, to normalcy. PMID- 10794044 TI - Does the intrauterine device carry the risk of immunity to sperm? AB - Intrauterine devices (IUDs) are the second most commonly used contraceptive method in the world. A number of mechanisms have been proposed by which IUDs could interfere with reproductive processes. In this study, the relationship between intrauterine devices and the risk of antisperm antibody (ASA) production in the absence of prior sensitization was investigated. Sixty-two IUD users (group 1) and 42 women with no contraceptive use as a control group (group 2) were included in the study. Six months after the IUD insertion, 4 women in group 1 and 2 women in group 2 with lower genital tract infections were excluded from the study. The sera of the remaining 58 women in group 1 and 40 women in group 2 were evaluated again for the presence of ASA. Twelve patients (20.7%) in group 1 and 12 patients (30.0%) in group 2 had ASA positivity. When we compared the ASA levels in the IUD group with those in the control group, there was no statistically significant difference (p>0.05). In summary, our data proved that copper-containing intrauterine devices in the absence of prior sensitization do not significantly affect immunity to sperm in sera of women. PMID- 10794045 TI - Two-year clinical experience with Nova-T 380, a novel copper-silver IUD. AB - In 1992, an open single-group phase III clinical trial was started at three centers to investigate the clinical performance of the high copper surface area Nova-T 380, a modification of Nova-T. This report presents the interim results of the first two years of use. A total of 400 women volunteers were enrolled in the study. The mean age was 31.4 years (SD 5.5) with a minimum of 18 and a maximum of 44 years. At the cut-off date, 259 women had passed the 24-month visit. Gross cumulative life-table rates at the end of the first and second years, respectively, calculated by the Kaplan-Meier method, were as follows: pregnancy 0.5 and 1.6, expulsion 1.6 and 2.8, bleeding 4.7 and 8.7, pain 1.3 and 2.3, removal for other medical reasons 1.7 and 3.9, planning pregnancy 1.1 and 6.0, removal for other personal reasons 0.5 and 1.5, per 100 users. No ectopic pregnancies or PIDs occurred. The continuation rates were 89.0 and 75.5 at 12 and 24 months, respectively. The first two-year performance of the Nova-T 380 was good. Bleeding problems were in the same range as with devices with smaller copper surface areas. No unexpected serious adverse events were encountered. PMID- 10794046 TI - Induced abortion: a method for birth control? AB - Despite the recent improvements in services and an increase in available funds, the expected increase in usage of effective contraceptive methods and decrease in number of induced abortions has not been seen in Turkey. This study investigates the causes of this situation and argues whether induced abortion is being used as a birth control method. Eight-five subjects were involved in the study, all of whom came to a family planning clinic for an induced abortion. Forty-seven (55.29%) of the participants had had at least one induced abortion prior to this. Eight-four (98.8%) had knowledge of at least one contraceptive method, and 80 (94.1%) had knowledge of at least one effective contraceptive method. Eighty (94.1%) participants had used some form of contraception at some time in their lives; 52 (61.17%) had used an effective contraceptive method at some time in their lives. When they became pregnant, 60 (70.6%) were using ineffective methods or were not using any contraceptive method. Both the participant and the spouse wanted the abortion in 68 (80%) cases. PMID- 10794047 TI - Communication and sexuality in a Nigerian community. AB - The traditional lack of interest in discussing sexuality creates a problem in doctor-patient communication, and this can affect patient management adversely. The dearth, ease or comfort in discussing sex was examined by a self-administered questionnaire to respondents, mainly medical students, nurses and paramedics, 20 70 years of age, who were not seeking treatment for sexual problems. The respondents were mainly of Igbo extraction from Eastern Nigeria. Factors considered include age, sex, religion, marital status, and education. The results show that 71.9% of all the respondents indicated that they would like to be able to discuss freely whereas 28.1% never really bothered; 40.9% of all the respondents could discuss sex with anybody whereas 59.1% could not; 75% in the married group discussed sex freely with their spouses or friends and 25% were unable to do so. Education showed a very significant influence on the ability to discuss sex freely. It is suggested that a systematic approach to education, especially sexual health education, may be a major way to combat the prevailing cultural inhibition. PMID- 10794048 TI - European multicenter study of natural family planning (1989-1995): efficacy and drop-out. The European Natural Family Planning Study Groups. AB - BACKGROUND: Effectiveness studies in natural family planning (NFP) published over the past 20 years have shown a wide range of contraceptive efficacy and acceptability. This seems to be due in part to different NFP methodologies. Consequently, we decided to carry out an effectiveness study in Europe to examine one group of the most widely spread NFP methods, the symptothermal methods. METHODS: Between 1989 and 1995, 15 NFP groups from 10 European countries participated in a prospective European multicentre study. This paper reports on 1328 women aged between 19 and 45 years and willing to participate for at least 12 cycles. Two types of symptothermal methods were mainly used, the symptothermal double-check methods (1046 women, 16865 cycles of exposure, 34 unintended pregnancies) and the symptothermal single-check methods (214 women, 1495 cycles of exposure, 13 unintended pregnancies). The study was an observational study with prospectively collected data. The pregnancy rates, drop-out rates and lost to-follow-up rates are presented separately for both subgroups according to the Kaplan-Meier method. RESULTS: For the double-check methods, there was an unintended pregnancy rate of 2.61% at the end of the first 12 cycles of use (standard error or SE 0.55%), a drop-out rate for difficulties or dissatisfaction of 3.9% (SE 0.69%) and a lost-to-follow-up rate of 3.1% (SE 0.62%). In the single check group, there was a total of 13 unintended pregnancies at the end of the first 12 cycles of study participation, giving an unintended pregnancy rate of 8.5% (SE 2.52%), a drop-out rate for difficulties or dissatisfaction of 3.0% (SE 1.76%) and a lost-to-follow-up rate of 23.4% (SE 4.35%). No pregnancy was observed in women over 40 years of age. Most pregnancies occurred because of deliberate unprotected intercourse in the fertile phase ('user failure'). CONCLUSIONS: The symptothermal double-check methods have proved to be effective family planning methods in Europe. The low drop-out-rate for difficulties or dissatisfaction with NFP shows the good acceptability. PMID- 10794049 TI - T-cell proliferation in vivo and the role of cytokines. AB - Unlike typical naive T cells, T cells with an activated (CD44hi) memory phenotype show a rapid rate of proliferation in vivo. The turnover of memory-phenotype CD8+ T cells can be considerably augmented by injecting mice with various compounds, including polyinosinic polycytidylic acid, lipopolysaccharide and immunostimulatory DNA (CpG DNA). Certain cytokines, notably type I (alpha, beta) interferons (IFN-I), have a similar effect. These agents appear to induce proliferation of CD44hi CD8+ cells in vivo by an indirect process involving production of effector cytokines, possibly interleukin-15, by antigen-presenting cells. Although none of the agents tested induces proliferation of naive phenotype T cells, IFN-I has the capacity to cause upregulation of surface markers on purified naive T cells. Depending upon the experimental conditions used, IFN-I can either inhibit or enhance primary responses of naive T cells to specific antigen. PMID- 10794050 TI - The organization of mature T-cell pools. AB - To deal with exogenous pathogens the peripheral T-cell compartment requires diverse repertoires (as those of naive cells) and efficient responses, the latter dependent on the persistence of memory cells. In the present work we show that (i) naive and memory cells differ in the type of interactions required for survival and division; (ii) they are segregated into independent ecological niches; (iii) that the size of each niche is controlled by independent homeostatic mechanisms; and (iv) that naive T cells do not have intrinsic life spans, surviving in the absence of thymus output but being continuously substituted by thymus export. The independent homeostatic regulation of the naive and memory T-cell pools guarantees the maintenance of versatile and efficient repertoires throughout life as well as the persistence of the naive T-cell pool after the thymus atrophies at puberty. PMID- 10794051 TI - A new theory of cytotoxic T-lymphocyte memory: implications for HIV treatment. AB - We use simple mathematical models to examine the dynamics of primary and secondary cytotoxic T-lymphocyte (CTL) responses to viral infections. In particular, we are interested in conditions required to resolve the infection and to protect the host upon secondary challenge. While protection against reinfection is only effective in a restricted set of circumstances, we find that resolution of the primary infection requires persistence of CTL precursors (GTLp), as well as a fast rate of activation of the CTLp. Since these are commonly the defining characteristics of CTL memory, we propose that CTL memory may have evolved in order to clear the virus during primary challenge. We show experimental data from lymphocytic choriomeningitis virus infection in mice, supporting our theory on CTL memory. We adapt our models to HIV and find that immune impairment during the primary phase of the infection may result in the failure to establish CTL memory which in turn leads to viral persistence. Based on our models we suggest conceptual treatment regimes which ensure establishment of CTL memory. This would allow the immune response to control HIV in the long term in the absence of continued therapy. PMID- 10794052 TI - B-cell memory and the persistence of antibody responses. AB - Antigens such as viral envelope proteins and bacterial exotoxins induce responses which result in the production of neutralizing antibody. These responses persist for years and provide highly efficient defence against reinfection. During these antibody responses a proportion of participating B cells mutate the genes that encode their immunoglobulin variable regions. This can increase the affinity of the antibody, but can also induce autoreactive B cells. Selection mechanisms operate which allow the cells with high affinity for the provoking antigen to persist, while other B cells recruited into the response die. PMID- 10794053 TI - Role of B cells in maintaining helper T-cell memory. AB - The cellular interactions involved in maintaining CD4+ T-cell memory have hitherto not been identified. In this report, we have investigated the role played by B cells in this process. We show that that long-lasting helper T-cell memory depends on the presence of B cells, but that direct antigen presentation by B cells is not required. These findings provide new insights into the mechanisms which underlie helper T-cell memory. They also suggest that the efficacy of future vaccines will depend critically on the inclusion of B- as well as T-cell epitopes. PMID- 10794054 TI - Memory in the B-cell compartment: antibody affinity maturation. AB - In the humoral arm of the immune system, the memory response is not only more quickly elicited and of greater magnitude than the primary response, but it is also different in quality. In the recall response to antigen, the antibodies produced are of higher affinity and of different isotype (typically immunoglobulin G rather than immunoglobulin M). This maturation rests on the antigen dependence of B-cell maturation and is effected by programmed genetic modifications of the immunoglobulin gene loci. Here we consider how the B-cell response to antigen depends on the affinity of the antigen receptor interaction. We also compare and draw parallels between the two processes, which underpin the generation of secondary-response antibodies: V gene somatic hypermutation and immunoglobulin heavy-chain class switching. PMID- 10794055 TI - The terminology problem for T cells: a discussion paper. AB - The school of thought that owes allegiance to Ludwig Wittgenstein teaches that language conditions perceptions. When we use the term 'cytotoxic T lymphocyte' or 'helper T cell' we tend to orientate our own thinking processes, and those of listeners or readers, down particular paths. Part of the problem is that we are often describing cell populations by functions that may either be a property of only a proportion of those that are being assayed, or are simply inferred from the expression of various cell-surface markers. The consequence can be a measure of confusion that might be avoided if we could communicate with greater clarity. Is it possible to achieve a better terminology that will be accepted generally? The following are some examples of why there may be some value in thinking about this. PMID- 10794056 TI - Memory CD8+ T cells in HIV infection. AB - Cytotoxic T lymphocytes (CTLs) play a central role in the control of persistent HIV infection in humans. The kinetics and general features of the CTL response are similar to those found during other persisting virus infections in humans. During chronic infection there are commonly between 0.1 and 1.0% of all CD8+ T cells in the blood that are specific for immunodominant virus epitopes, as measured by HLA class I peptide tetramers. These figures are greatly in excess of the numbers found by limiting dilution assays; the discrepancy may arise because in the latter assay, CTLs have to divide many times to be detected and many of the HIV-specific CD8+ T cells circulating in infected persons may be incapable of further division. Many tetramer-positive T cells make interferon-gamma, beta chemokines and perforin, so are probably functional. It is not known how fast these T cells turn over, but in the absence of antigen they decay in number. Impairment of CTL replacement, because CD4+ T helper cells are depleted by HIV infection, may play a major role in the development of AIDS. PMID- 10794057 TI - On immunological memory. AB - Immunological memory may not represent a special characteristic of lymphocytes but simply reflect low-level responses driven by antigen that is re-encountered or persists within the host. T-cell memory is important to control persistent infections within the individual host and cannot be transmitted to offspring because of MHC polymorphism and MHC-restricted T-cell recognition. In contrast, antibody memory is transmissible from mother to offspring and may function essentially to protect offspring during the phase of physiological immuno incompetence before, at and shortly after birth. This physiological immuno incompetence is a result of MHC polymorphism and the dangers of the graft-versus host and host-versus-graft reaction between mother and embryo, which necessitate immunosuppression of the mother and immuno-incompetence of the offspring. One may argue therefore that immunological memory of transmissible immunological experience is the basis on which MHC-restricted T-cell recognition could develop or coevolve. PMID- 10794058 TI - Antiviral CD4 and CD8 T-cell memory: differences in the size of the response and activation requirements. AB - Following acute lymphocytic choriomeningitis virus (LCMV) infection, there is a potent antiviral CD8 T-cell response that eliminates the infection. This initial CD8 T-cell response is followed by a period of memory during which elevated numbers of virus-specific CD8 T cells remain in the mouse. CD4 T cells are also activated after LCMV infection, but relatively less is known about the magnitude and duration of the CD4 response. In this study, we used intracellular staining for interferon-gamma to measure both CD4 and CD8 responses in the same mice at the single cell level. After LCMV infection, there was an increase in the number of activated CD4 T cells and an associated increase in the number of virus specific CD4 T cells. At the peak of this expansion phase, the frequency of virus specific CD4 T cells was 1 in 20 (0.5-1.0 x 10(6) per spleen). Like the CD8 response, long-term CD4 memory could be found up to a year after the infection with frequencies of approximately 1 in 260 (0.5-1.5 x 10(5) per spleen). However, the magnitude of virus-specific CD8 T cells was greater than virus-specific CD4 T cells during all phases of the immune response (expansion, death, and memory). At day 8, there were 20- to 35-fold more virus-specific CD8 T cells than CD4 T cells. This initial difference in cell number lasted into the memory phase as there remained a ten- to 20-fold difference in the CD8 and CD4 responses. These results highlight the importance of the expansion phase in determining the size of the memory T-cell pool. In addition to the difference in the magnitude, the activation requirements of CD8 and CD4 T-cell responses were different: CD8 T responses were not affected by blockade of CD40-CD40 ligand interaction whereas CD4 responses were reduced 90%. So while there is long-term memory in both the CD8 and CD4 compartments, the rules regulating the activation of CD8 and CD4 T cells and the overall magnitude of the responses are different. PMID- 10794059 TI - Immunological memory and acquired immunodeficiency syndrome pathogenesis. AB - Infection with the human immunodeficiency virus results in profound perturbations in immunological memory, ultimately resulting in increased susceptibility to opportunistic infections and acquired immunodeficiency syndrome (AIDS). We have used rhesus macaques infected with the simian immunodeficiency virus (SIV) as a model to understand better the effects of AIDS virus infection on immunological memory. Acute infection with SIV resulted in significant deficits in CD4+ helper responses to cytomegalovirus (CMV) as well as CMV-specific cytotoxic T-lymphocyte and neutralizing antibody responses. Reactivation of CMV was associated with high levels of SIV replication and suppression of both T-helper and cytotoxic responses to CMV. We have also studied the effects of SIV infection on T-cell turnover in non-human primates. T-cell turnover was evaluated using the nucleoside analogue bromodeoxyuridine (BrdU) in combination with five-colour flow cytometric analysis. T cells in normal animals turned over at relatively rapid rates, with memory cells turning over more quickly than naive cells. In SIV infected animals, the labelling and elimination rates of both CD4+ and CD8+ BrdU labelled cells were increased by two- to threefold compared with normal controls. Further analysis of immunological memory in non-human primates should offer the opportunity to extend immunological insights from murine models to the pathogenesis and prevention of AIDS. PMID- 10794061 TI - Differences in the regulation of CD4 and CD8 T-cell clones during immune responses. AB - The functional units of immune response are lymphocyte clones. Analysis of lymphocyte life span in vivo shows that the overall turnover of CD4 and CD8 lymphocytes does not differ greatly. Recently, molecular methods have been developed which allow a global analysis of T-cell clones responding to an antigen in vivo. We have used a sensitive, modified heteroduplex analysis to follow T cell clones responding to Epstein-Barr virus in acute infectious mononucleosis (AIM). Strikingly, all the many large clones detected in freshly isolated AIM blood were found within the CD8 fraction. CD4 clonal populations responding to the soluble recall antigen tetanus toxoid could only be detected after in vitro re-stimulation. These data imply that CD4 responses may be more polyclonal than those of CD8 cells and that the size of CD4 clones is more tightly regulated. Several molecular mechanisms may contribute to this. Up-regulation of telomerase allows very large expansions of CD8 cells to occur without exhaustion of proliferative capacity. PMID- 10794060 TI - T-cell memory: lessons from Epstein-Barr virus infection in man. AB - Epstein-Barr virus offers an ideal opportunity to follow the human T-cell response to a virus infection over time from its acute primary phase, as seen in infectious mononucleosis patients, into the memory phase that accompanies life long virus persistence. Here we review recent evidence on the development and maturation of cytotoxic T-cell memory using this viral system. PMID- 10794062 TI - CD4 T-cell memory can persist in the absence of class II. AB - To understand how memory CD4 T cells are generated we have re-examined the requirements for continuing antigen stimulation in the generation and persistence of this population. We find that specific antigen is only required for a short period during the activation of naive CD4 T cells and is not required for memory generation from activated CD4 T cells or for persistence of resting memory cells generated by transfer of activated CD4 to adoptive hosts. Moreover, transfer of activated CD4 T cells to class-II-deficient hosts, indicates that TcR-class II major histocompatibility interaction is also unnecessary for either the transition from activated CD4 T cell to resting memory cells or for persistence over an eight-week period. Thus the signals regulating generation and maintenance of memory are fundamentally different from those which regulate the expansion of effector CD4 T-cell populations which include antigen itself and the CD4 T-cell autocrine cytokines induced by antigen. PMID- 10794063 TI - Does melatonin modulate beta-endorphin, corticosterone, and pain threshold? AB - Converging lines of evidence suggest that the pineal hormone, melatonin, may regulate changes in pain threshold by modulating fluctuations in opioid receptor expression and levels of beta-endorphin (beta-END). This study investigated whether the circadian oscillation in plasma melatonin is involved in the modulation of plasma beta-END immunoreactivity (beta-END-ir), and whether fluctuations in pain threshold measured using the hotplate test are contingent upon the fluctuation of these two hormones in Rattus Norvegicus. The role of melatonin was explored using light-induced functional pinealectomy (LFPX) to suppress nocturnal melatonin release. Pinealectomized rats were found to have significantly elevated levels of beta-END-ir compared to control animals at both photophase (398 +/- 89 pg/ml versus 180 +/- 23 pg/ml) and scotophase (373 +/- 45 pg/ml versus 203 +/- 20 pg/ml) test-periods, thus supporting the putative melatonin-opioid axis. Similarly, latency to pain threshold of LFPX rats was significantly longer when compared to control animals at photophase (7.3 +/- 1.4 sec versus 4.8 +/- 0.7 sec) and scotophase (6.3 +/- 0.7 sec versus 5.1 +/- 0.7 sec). Previous studies have produced conflicting data regarding the role of the pineal system in modulating levels of corticosterone (CORT). We observed a moderate, but non-significant, increase in the CORT concentration of LFPX rats during the photophase test period. PMID- 10794064 TI - Chronic low dose ethanol intake: biochemical characterization of liver mitochondria in rats. AB - Liver mitochondria were isolated from male rats exposed for 2 months to low doses of ethanol (3% v/v in drinking water), a condition not associated with tolerance or dependence. The results show no significant changes in the content of reduced or oxidized glutathione in the liver mitochondria of ethanol treated rats with respect to controls. However, a slight but significant increase in lipid peroxidation, accompanied by an increased content of oxidized proteins, was found in ethanol exposed animals. Mitochondrial content of cytochrome complexes was not significantly affected by ethanol intake. The specific enzymatic activity of cytochrome oxidase showed, however, a significant decrease in ethanol-treated rats. The slight mitochondrial alterations found in the liver of rats exposed chronically to low doses of ethanol might represent the beginning of a more extensive damage previously observed in rats exposed to high doses of this substance. PMID- 10794066 TI - Body distribution of 3H-labelled dalargin bound to poly(butyl cyanoacrylate) nanoparticles after i.v. injections to mice. AB - The blood-brain barrier (BBB) limits the penetration of substances into the brain. Because many drugs, particularly peptides, therefore can not be delivered to the brain, carrier systems were developed to overcome this problem. In earlier studies we demonstrated central analgesic effects of a peptide, dalargin (dal), after systemic administration when this substance was bound onto the surface of polybutylcyanoacrylate nanoparticles and coated with polysorbate 80 but not when it was given alone. The aim of the present study was to investigate the body distribution of 3H-labelled dal bound to nanoparticles compared to unbound dal after i.v. injection in mice. The radioactivity in several tissues, including the brain, was separated in subcellular preparations and was measured after a single i.v. injection over time. Dal radioactivity level in brain preparations was 3 times higher when the drug was bound to nanoparticles whereas the first pass pathway in liver was reduced. The results support previous data that nanoparticles can be used to transport peptides across the BBB. PMID- 10794065 TI - Unilateral 6-hydroxydopamine lesions of nigrostriatal dopaminergic neurons increased CB1 receptor mRNA levels in the caudate-putamen. AB - It has been recently suggested that the effects of cannabinoids on motor behavior might be different in rats with lesions of nigrostriatal dopaminergic neurons than in controls. In the present study, we examined the possible alteration in the status of cannabinoid CB1 receptors in the basal ganglia of rats with unilateral lesions of those neurons caused by 6-hydroxydopamine. We used two different experimental groups depending on the duration of the period of recovery after the lesion, and comparisons were done between the lesioned and nonlesioned sides at the level of the basal ganglia. Both groups of lesioned rats exhibited a similar marked reduction in tyrosine hydroxylase (TH)-mRNA levels, measured by in situ hybridization, in the substantia nigra of the lesioned side. In the same way, lesioned rats exhibited the characteristic rotational behavior after a single injection of apomorphine and the intensity of this rotation was stable at the two times analyzed after the lesion. Also as expected, lesioned rats exhibited an increase in proenkephalin mRNA levels in the caudate-putamen, whereas mRNA levels of substance P decreased, although differences between the two times of recovery analyzed were observed in this case. We did not find any significant changes in CB1 receptor binding, measured by [3H]WIN-55,212,2 autoradiography, or in the activation of signal transduction mechanisms, measured by WIN-55,212,2-stimulated [35S]GTPgammaS binding autoradiography, between the lesioned and nonlesioned sides at the level of the lateral caudate-putamen, globus pallidus and substantia nigra in both groups of lesioned rats. However, we found a significant increase in levels of CB1 receptor-mRNA transcripts, measured by in situ hybridization, in the lesioned side in both the lateral and medial caudate-putamen. This occurred 7-10 weeks after the lesion, but the increase was markedly waned after 17-18 weeks. In summary, the unilateral 6-hydroxydopamine lesion of nigrostriatal dopaminergic neurons originated a marked increase in CB1 receptor-mRNA levels in cell bodies of striatal efferent neurons, although accompanied by no changes in CB1 receptor binding and activation of signal transduction mechanisms. This supports a critical role for dopamine in the control of CB1 receptor gene expression. However, the magnitude of the effect significantly waned as a function of the duration of the period after lesion. PMID- 10794067 TI - Protective effects of melatonin against ischemia-reperfusion injury in the isolated rat heart. AB - There has been increased interest in melatonin recently, since it was shown to be a potent scavenger of toxic free radicals. Melatonin has been found to be effective in protecting against pathological states due to reactive oxygen species release. The present study was performed in order to determine whether melatonin or 5-methoxy-carbonylamino-N-acetyl-tryptamine (5-MCA-NAT), a structurally related indole compound, protect against ischemia-reperfusion injury in the isolated rat heart. Wistar rats were treated in vivo with either melatonin (1 or 10 mg/kg, i.p.) or 5-MCA-NAT (10 mg/kg, i.p.) or their vehicle, 30 min before their hearts were excised and perfused according to the Langendorff technique. Two different protocols were then applied. In the first one, a regional ischemia (5 min)-reperfusion (30 min) sequence was performed in order to record incidence and duration of reperfusion arrhythmias. In the second one, infarct size was assessed after a regional ischemia (30 min)-reperfusion (120 min) sequence. Results show a spectacular protection against ischemia-reperfusion injuries (on arrhythmias as well as on infarct size) in rats pre-treated with 10 mg/kg of melatonin or 5-MCA-NAT. In conclusion, both melatonin and its structural analog, 5-MCA-NAT, appear to confer protection against ischemia-reperfusion injury in the isolated rat heart. This observation suggests that melatonin could have a potential clinical application in the treatment of myocardial ischemia, even if the mechanisms underlying this protection remain to be determined. PMID- 10794068 TI - G-proteins in kidneys of spontaneously hypertensive rats. AB - G(s alpha)-, total G(i alpha)- and G(q/11alpha)-protein concentrations were investigated by quantitative immunoblotting in membranes of total kidney, renal cortex and medulla as well as in cortical tubules and glomeruli of Spontaneously Hypertensive Rats (SHR) and normotensive Wistar Kyoto rats (WKY), aged 5 weeks, 3 or 8 months. We found that total kidney of 5 week old SHR possess less G(s alpha) , G(i alpha)- and G(q/11alpha)-proteins than controls. For G(s alpha)-proteins, differences found in total kidney were mirrored both in cortex (tubules and glomeruli) and in medulla. Decreased G(i alpha)-concentrations were accompanied by lower tubular but higher glomerular levels, while medullar levels were also increased. Decreased G(q/11alpha)-concentrations were reflected in decreased glomerular and medullary concentrations. Kidneys of 3 month old SHR and WKY possessed similar concentrations of all G(alpha)-species. In 8 month old SHR similar G(i alpha)-, but decreased G(s alpha)-and G(q/11alpha)-concentrations were observed. The G(s alpha)-decrease was reflected in cortex and medulla, the G(q/11alpha)-decrease in the medulla. We conclude that the main strain-related differences in G(alpha)-concentrations are seen in prehypertensive SHR. PMID- 10794069 TI - Effect of two high-oleic oils on the liver lipid composition of spontaneously hypertensive rats. AB - Despite having similar fatty acid composition and plasma lipid composition after ingestion, olive oil, but not high-oleic sunflower oil (HOSO), is capable of reducing blood pressure. HOSO contains mainly triolein, whereas olive oil contains important amounts of dioleoyl-palmitoyl-glycerol. In order to see if its different triacylglycerol (TAG) composition could be related to the hypotensive effect of olive oil, Spontaneously Hypertensive Rats (SHR) were fed with HOSO and olive oil-rich diets. Liver lipid composition was determined. Total lipid, fatty acid and TAG composition was analyzed. Rats fed olive oil (67.24 +/- 4.23) were observed to retain more dioleoyl-acyl-glycerol species in their liver than those fed HOSO (56.6 +/- 3.95), specially triolein (20.69 +/- 1.77 olive oil, vs. 12.54 +/- 1.97 HOSO), in spite of its lower content of this TAG. On the contrary, rats consuming HOSO had higher amounts of dilinoleoyl-acyl-glycerol species (9.26 +/- 1.57 HOSO, vs.4.02 +/- 0.90 olive oil). In conclusion, olive oil provided a more beneficial TAG profile in the liver of SHR rats than HOSO, probably due to the differences in the TAG composition of both oils. PMID- 10794070 TI - Estrogen supplement prevents the calcium hypersensitivity of cardiac myofilaments in ovariectomized rats. AB - Our previous biochemical and mechanical studies have demonstrated an increase in Ca2+ sensitivity of cardiac myofilaments in ovariectomized rats. To test whether the body weight gain associated with ovariectomy contributed some effects to the changes in myofibrillar functions, the relations of pCa (-log Ca2+ molar concentration) to actomyosin adenosine triphosphatase (ATPase) activity of isolated myofibrillar preparations from 10-week pair-fed ovariectomized rats were compared with those from sham-operated controls. Despite similar body weights, the maximum myofibrillar ATPase activity was significantly lower in pair-fed ovariectomized rats as compared to that of sham-operated controls. In addition, the pCa-actomyosin ATPase relationship of pair-fed ovariectomized hearts still demonstrated a significant leftward shift in pCa50 (-log half-maximally Ca2+ activation) from that of sham-operated controls. To find out which hormone was responsible for the observed increase in myofibrillar Ca2+ sensitivity, different sex hormone supplemental regimens were administered to ovariectomized rats. Subcutaneous injection of estrogen (5 microg/rat) or estrogen plus progesterone (1 mg/rat) three times a week could effectively prevent the changes in body weight, heart weight, and uterine weight of the ovariectomized animals. Moreover, supplements of either estrogen or progesterone could prevent a decrease in maximum ATPase activity. In contrast, only the estrogen replacement could abolish the Ca2+ hypersensitivity of the myofilaments in these ovariectomized rats. These results suggest differential cardio-regulatory effects of ovarian sex hormones on the Ca2+ activation of the myofilaments. PMID- 10794072 TI - Iron release and oxidant damage in human myoblasts by divicine. AB - Divicine is an aglycone derived from vicine, a glucosidic compound contained in fava beans (Vicia faba major or broad beans). In this study, we investigated the effect of divicine on cultured human myoblasts from normal subjects, in order to see if the drug may induce signs of oxidant stress in these cells. Myoblasts incubated 24 hours in the presence of 1 mM divicine, showed an increase of carbonyl groups and 4-hydroxynonenal (4-HNE) bound to cell proteins, as well as a significant release of iron and lactate dehydrogenase in the culture medium. Desferrioxamine (DFO), an iron chelator, significantly prevented protein oxidation and formation 4-HNE adducts. Our results can be interpreted as indicating that divicine autooxidizes both at extracellular level and into myoblasts thus inducing the release of free iron, which initiates oxidation of cellular proteins and lipids. DFO protects the cells by subtracting the free iron both at intracellular and extracellular level. PMID- 10794071 TI - Differential effect of clofibrate on acetyl-CoA carboxylase mRNA level in rat white and brown adipose tissue. AB - Regulation of some lipogenic enzyme gene expression by clofibrate was studied in rat white and brown adipose tissue. In white adipose tissue the drug administration for 14 days to rats resulted in the increase in acetyl-CoA carboxylase, ATP-citrate lyase, and glucose 6-phosphate dehydrogenase mRNA levels. Opposing effect of clofibrate on the acetyl-CoA carboxylase, ATP-citrate lyase, and glucose 6-phosphate dehydrogenase mRNA levels was found in brown adipose tissue. These data indicate a tissue specificity of clofibrate action on lipogenic enzyme gene expression. The results presented in this paper provide further evidence that hypolipidaemia caused by the treatment with clofibrate cannot be related to the inhibition of fatty acid synthesis in white adipose tissue in rat. PMID- 10794073 TI - Effects of repeated stress on expression of interleukin-6 (IL-6) and IL-6 receptor mRNAs in rat hypothalamus and midbrain. AB - We examined the effects of single and repeated stress on the expression of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) mRNAs in the rat midbrain and hypothalamus using reverse transcriptase-polymerase chain reaction (RT-PCR). Following a single episode of restraint stress for 4 hours (1R) or 4 hours per day on two (2R) or three (3R) consecutive days, the hypothalamus and midbrain were removed immediately and the levels of IL-6 and IL-6R mRNAs in both regions were determined. Regional differences in stress-related changes in mRNA levels were noted. The expression of IL-6 mRNA in the hypothalamus did not change in 1R group but decreased in 2R and 3R groups. The expression of IL-6R mRNA in the same region significantly diminished in all groups. In the midbrain, the expression of IL-6 mRNA increased in 1R group and decreased in 2R and 3R, while the expression of IL-6R mRNA significantly diminished in 1R and 3R groups but was not different from control in 2R group. Our findings indicate that repeated stress in rats produce changes in IL-6 and IL-6R mRNAs in the midbrain and hypothalamus that are different than those of a single stress episode. PMID- 10794074 TI - Nociceptin receptor activation produces nitric oxide-mediated systemic hypotension. AB - The purpose of the present study was to investigate the effects of L-N5-(1 iminoethyl)ornithine hydrochloride (L-NIO), an inhibitor of nitric oxide (NO) formation, and [Phe1-psi(CH2NH)-Gly2]Nociceptin(1-13)-NH2 (Phe-NOC), a nociceptin receptor antagonist, on the systemic vasodepressor response to nociceptin in the anesthetized rat. The systemic vasodepressor response to bolus intravenous (i.v.) injections of nociceptin was significantly reduced by L-NIO and Phe-NOC. The present data suggest activation of nociceptin receptors dilates the systemic vascular bed through a NO-dependent pathway. These data also demonstrate Phe-NOC is an efficacious and selective nociceptin receptor antagonist in vivo. PMID- 10794076 TI - Chitin oligosaccharides as candidate patterning agents in zebrafish embryogenesis. AB - In this work we investigate the possible function of N-acetyl chitooligosaccharides (NACOs) produced during zebrafish (Danio rerio) development. First, we show that NACOs are synthesized in vivo during early embryogenesis in the zebrafish. Second, we demonstrate that injection of a pure bacterial chitinase into one-cell stage embryos elicits developmental defects in which the posterior trunk and tail of developing embryo are severely affected. In addition, an endogenous chitinase activity detected both intra- and extracellularly is described, suggesting that cells may secrete it into the extracellular space. Moreover, this compartmentalization appears to be functionally relevant as inhibition of the extracellular, but not the intracellular, endogenous chitinase activity causes morphological defects similar to those seen in embryos injected with chitinase 63. Finally, analysis of the expression of the zebrafish ZDG42 gene, which has been suggested to be involved in synthesis of NACOs, is described. Transcripts are detected from late blastula stage, during gastrulation, and move as an anterior-posterior wave of expression in adaxial mesoderm during somitogenesis. PMID- 10794075 TI - X chromosome inactivation revealed by the X-linked lacZ transgene activity in periimplantation mouse embryos. AB - Using H253 mouse stock harboring X-linked HMG-lacZ transgene, we examined X chromosome inactivation patterns in sectioned early female embryos. X-gal staining patterns were generally consistent with the paternal X inactivation in the trophectoderm and the primitive endoderm cell lineages and random inactivation in the epiblast lineages. The occurrence of embryonic visceral endoderm cells apparently at variance with the paternal X chromosome inactivation in 7.5 dpc embryos was explained by the replacement of visceral endoderm cells with cells of epiblast origin. The frequency of cells negative for X-gal staining in 4.5-5.5 dpc XmXp* embryos fluctuated considerably especially in the extraembryonic ectoderm and the primitive endoderm, whereas it was less variable in the embryonic ectoderm. We could not, however, determine whether it is a normal phenomenon revealed for the first time by the use of HMG-lacZ transgene or an abnormality caused by the multicopy transgene. PMID- 10794077 TI - Requirement of protamine for maintaining nuclear condensation of medaka (Oryzias latipes) spermatozoa shed into water but not for promoting nuclear condensation during spermatogenesis. AB - Protamine is an arginine-rich basic protein found in the sperm nuclei of many vertebrates, but its actual roles in spermatozoa remain to be elucidated. In this study, we investigated the physiological roles of protamine by examining protamine-less spermatozoa produced in vitro in the presence of the transcriptional inhibitor actinomycin D. Even under inhibited transcription, medaka spermatocytes underwent meiosis and differentiated into spermatozoa with a condensed nucleus and an elongated flagellum. Using a newly produced anti-medaka protamine antibody, we confirmed the absence of protamine protein in the spermatozoa differentiated in the presence of actinomycin D. These findings clearly indicate that sperm nuclear condensation in medaka is independent of protamine. Since medaka spermatozoa are shed into water upon natural fertilization, we also investigated the roles of protamine by comparing the differences between the nuclear morphology of protamine-equipped and protamine less spermatozoa immersed in water. The nuclei without protamine more rapidly swelled than did those with protamine and completely broke down within 10 min, whereas more than 80% of the sperm nuclei with protamine resisted the disruption under similar conditions. These findings strongly suggest that a physiological role of protamine in medaka spermatozoa is to protect the ejaculated spermatozoa against the disruption by low osmotic pressure until arrival at the eggs for successful fertilization. PMID- 10794078 TI - Colony-stimulating factor-1 (CSF-1) rescues osteoblast attachment, survival and sorting of beta-actin mRNA in the toothless (tl-osteopetrotic) mutation in the rat. AB - We have shown that in the osteopetrotic rat mutation toothless (tl) osteoblasts are absent from older bone surfaces in mutants and that mutant osteoblasts in vivo lack the prominent stress fiber bundles polarized along bone surfaces in osteoblasts from normal littermates. Our recent data demonstrate that in normal osteoblasts in vitro beta- and gamma-actin mRNAs have different, characteristic intracellular distributions and that tl (mutant) osteoblasts fail to differentially sort these mRNAs. Because bone resorption and formation are highly interdependent and injections of CSF-1, a growth factor, increase bone resorption and growth in tl rats, we examined the effects of CSF-1 treatment on osteoblast survival and ultrastructure in vivo and ability to sort actin mRNAs in vitro. Neonatal CSF-1 treatment of mutants restores osteoblasts on older bone surfaces, normalizes the intracellular distribution of stress fibers in osteoblasts in vivo and promotes normal sorting of beta-actin mRNA in mutant osteoblasts in vitro without normalizing gamma-actin distribution. These data suggest the beta- and gamma-actin mRNAs in osteoblasts are sorted by different mechanisms and that the differential sorting of beta-actin mRNA is related to the characteristic polarization of stress fibers in osteoblasts and their survival on bone surfaces. This experimental system can be used to explore the relationships and regulation of these aspects of cell and tissue biology. PMID- 10794079 TI - Endocytosis and transcytosis in growing astrocytes in primary culture. Possible implications in neural development. AB - Endocytosis constitutes an essential process in the regulation of the expression of cell surface molecules and receptors and, therefore, could participate in the neural-glial interactions occurring during brain development. However, the relationship between endocytic pathways in astroglial cells under physiological and pathological conditions remains poorly understood. We analyzed the endocytosis and transcytosis processes in growing astrocytes and the possible effect of ethanol on these processes. Evidence demonstrates that ethanol affects endocytosis in the liver and we showed that ethanol exposure during brain development alters astroglial development changing plasma membrane receptors and surface glycoprotein composition. To study these processes we use several markers for receptor-mediated endocytosis, fluid phase endocytosis and non-specific endocytosis. These markers were labeled for fluorescence microscopy and electron microscopy. 125I-BSA was used to study the effect of ethanol on the internalization and recycling of this macromolecule. The distribution of several proteins involved in endocytosis (caveolin, clathrin, rab5 and beta-COP) was analyzed using immunofluorescence, immunoelectron microscopy and immunoblotting. Our results indicate that growing astrocytes have a developed endocytic system mainly composed of caveolae, clathrin coated pits and vesicles, tubulo-vesicular and spheric endosomes, multivesicular bodies and lysosomes. Ethanol exposure induces a fragmentation of tubular endosomes, decreases the internalization of 125I-BSA, alters the processing of internalized BSA, and decreases the levels of caveolin, clathrin, rab5 and beta-COP. These results indicate that ethanol alters the endocytosis and transcytosis processes and impairs protein trafficking in astrocytes, which could perturb astrocyte surface expression of molecules involved in neuronal migration and maturation during brain development. PMID- 10794080 TI - Rapid disappearance of the medial epithelial seam during palatal fusion occurs by multifocal breakdown that is preceded by expression of alpha smooth muscle actin in the epithelium. AB - Breakdown of the medial epithelial seam (MES) is essential to allow bridging of the mesenchyme during palatal fusion. Evidence exists for three mechanisms for this breakdown that are incompatible at the level of individual cells in the seam. To determine if breakdown of the seam was regionally restricted, 3 dimensional reconstructions were generated using volume rendering software from 1 micron serial sections in the sagittal plane of rat palates fixed during the process of fusion. The earliest break detected in electron micrographs was cell separation and in reconstructions was a discrete defect, with a rounded outline, nearer to the nasal than to the oral margin of the seam. Further breakdown produced a pattern of rounded defects along the nasal margin of the seam resulting in interconnected columns of cells preferentially attached to the oral epithelium. Computer generated slicing of reconstructed seams showed that groups of cells evident in cross-sections as islands at this stage of breakdown of the MES could be artifacts. Unequivocal islands of epithelial cells formed later in fusion had a rounded outline, an incomplete basal lamina and a halo of cells containing phagocytosed apoptotic debris. The pattern of breakdown indicated that the MES breaks down under tension. Laser confocal microscopy of sections and whole-mounts of palates demonstrated alpha-smooth muscle actin preferentially localized in the epithelial cells of the palatal shelves immediately before and during formation of the seam. Expression in epithelial cells of the isoform of actin normally restricted to smooth muscle cells engaged in tonic contraction supported an interpretation that the epithelial cells of the seam may be capable of generating tension during the palatal fusion event. PMID- 10794081 TI - The presence of rudimentary odontogenic structures in the mouse embryonic mandible requires reinterpretation of developmental control of first lower molar histomorphogenesis. AB - In the mouse embryonic maxilla, rudimentary tooth primordia have been identified, which can be mistaken for the first upper molar. In order to determine whether such a situation might exist in the lower jaw as well, tooth development was investigated in the mouse mandibular cheek region during ED 12.5-15.0. A combination of histology, morphometry and computer-aided 3D reconstructions demonstrated the existence of rudimentary dental structures, whose gradual appearance and regression was associated with the segmental progress of odontogenesis along the mesio-distal axis of the jaw: 1) At ED 12.5, the mesial segment (MS) was the most prominent part of the dental epithelial invagination. It included an asymmetrically budding dental lamina. The MS, although generally mistaken for the lower first molar (M1, primordium, regressed and did not finally participate in M1 cap formation. 2) At ED 13.5, a wide dental bud (called segment R2) appeared distally to the MS. Although the R2 segment transiently represented the predominant part of the dental epithelium at ED13.5, it participated only in the formation of the mesial end of the M1 cap. 3) The top of the R2 segment at ED13.5 was not the precursor of the enamel knot (EK), contrary to what has been assumed. 4) The central segment of the M1 cap as well as the EK developed later and distally to the R2 segment. 5) Time-space specific apoptosis correlated with the retardation in growth of the R2 segment as well as with strong regressive changes in the epithelium situated mesially to it. These highlight the need to reinterpret current molecular data on early M1 development in the mouse in order to correlate the expression of signalling molecules with specific morphogenetic events in the appropriate antemolar or molar segments of the embryonic mandible. PMID- 10794083 TI - The cytoskeletal effector xPAK1 is expressed during both ear and lateral line development in Xenopus. AB - xPAK1, a probable effector of stress activated MAP-kinase SAPK1/JNK activation and cytoskeletal dynamics, was found to be ubiquitously expressed within the Xenopus laevis ear and lateral line system during the development and differentiation of these organs. xPAK1 expression was very strong in the otic placode from its condensation, and expression continued in the otic vesicle up until stage 35/36, after which it abruptly ceased. At stage 29/30 expression occurred specifically in the epithelium of the otic vesicle, which includes the prospective sensorial epithelium. Expression of xPAK1 was also observed in the lateral line system from stage 35/36, at which stage the lateral line primordia have begun to migrate from the region of the otic vesicle. Lateral line expression continued at least until stage 37/38, at which time xPAK1 was noted in association with the differentiating lateral line organs. To our knowledge, xPAK1 is the first ubiquitous lateral line marker that is also expressed in the ear. In the context of previous studies, our data suggest that xPAK1 either plays a role in the differentiation of the mechano-sensors of the auditory system or in the formation of the otic vesicle epithelium and the lateral line primordia. PMID- 10794082 TI - The meiotic specific synaptonemal complex protein SCP3 is expressed by female and male primordial germ cells of the mouse embryo. AB - The synaptonemal complex proteins SCP3 and SCP1 are components of the synaptonemal complex, a meiosis-specific protein structure essential for synapsis of homologous chromosomes. Using polyclonal antibodies raised against SCPs of rat testis, we have studied the expression of these proteins in embryonic germ cells of the mouse embryo using immunohistochemistry and immunoblotting. This investigation provided the first description of the sequential appearance of SCP3 and SCP1 during the different stages of the female meiosis in the mouse. Most importantly, we found that also male primordial germ cells express SCP3 for a short time before undergoing G1 arrest. This strongly supports the hypothesis that primordial germ cells are programmed to enter meiosis unrespective of the sex and that foetal testis produces a factor that inhibits such programme. PMID- 10794084 TI - Intestinal fatty acid binding protein gene expression reveals the cephalocaudal patterning during zebrafish gut morphogenesis. AB - Intracellular fatty acid-binding proteins (FABPs) are small and highly conserved cytoplasmic proteins that bind long-chain fatty acids and other hydrophobic ligands. We have examined, as a model for studying intestinal epithelial cell differentiation, the cell-specific and spatio-temporal expression of intestinal fatty acid-binding protein (i-fabp) gene during zebrafish larval development. After molecular cloning of zebrafish I-FABP cDNA, whole-mount in situ hybridization analysis revealed that i-fabp is expressed in the intestinal tube around day 3 postfertilization. By day 4, highest level of i-fabp transcript is encountered in the proximal columnar epithelium. From day 5 onwards, i-fabp is strongly expressed in the anterior intestine and its rostral expansion, slightly expressed in the esophagus mucosa and rectum, while no mRNA could be detected in the posterior intestine. Therefore, the regional differentiation of the intestine precedes first feeding and complete yolk resorption. I-fabp expression in the anterior intestine of the fed larvae is correlated with an intracellular storage of lipid droplets in the enterocytes and the massive synthesis of very low density lipoprotein particles. In conclusion, the cephalocaudal expression pattern of i-fabp demarcates early during zebrafish gut morphogenesis the anterior fat absorbing to posterior cells of the intestine. This gene could be used as a marker for screening for mutations that affect the events of intestinal epithelial differentiation, cephalocaudal patterning, and asymmetric gut looping morphogenesis. PMID- 10794085 TI - International acceptance of blood and blood products. AB - International agreement on the standardisation of blood and blood products is of crucial importance in helping to ensure that those biological medicines are safe and effective. International efforts to establish norms for the acceptability of biological products are accomplished primarily through the WHO, and many international reference materials needed for calibration, such as standards for blood grouping and hepatitis C virus RNA have been established by WHO. In addition to coordinating the development of physical standards, WHO also has been the major player in attempting to generate an international consensus on the basic criteria for the acceptability of biological products. However, requirements for safe blood and blood products are fluid because new risks or new infectious agents continue to be identified. For all those reasons, worldwide harmonization of the quality of blood and blood products remains an elusive goal. However, opportunities to move forward exist, and WHO is in a good position to act as a catalyst for progress. PMID- 10794086 TI - Immunobiology of persistent blood-borne viral infections. AB - Host-virus interactions have co-evolved to play an interactive role in the pathogenesis of viral infections and their disease outcome. Host responses to viral infections, including the cell-mediated and humoral immune responses, have been a subject of intensive research in virology and immunology. Definition of specific cellular receptors for cellular entry of the agents, the rates of their intracellular viral replication, the rates of turnover of circulating virions, persistence of viral infection possibly due to inadequate immune responses, and continued formation of circulating immune complexes provide the framework for our current understanding of the immunopathology of virally induced disease. Among the multiple blood-borne viruses (BBV) transmissible through transfusion, hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency viruses (HIV-1/-2) are relatively more important than several other viruses. Not only do they establish asymptomatic persistent infections with occasional oncogenic sequelae, but they also cause significant morbidity and mortality when transmitted through transfusion of blood and blood products. Molecular characterization of these agents and their in vitro inactivation and removal from blood have become key issues in contemporary transfusion safety since the advent of AIDS. Because many of the BBV are associated with white blood cells that have no therapeutic benefit in haemotherapy, simple filtration-removal of leukocytes from donated blood confers a dual benefit of immunological and virological safety in transfusion medicine. PMID- 10794089 TI - A safer plasma supply from remunerated donors--"The Immuno/Community Bio Resources experiment". AB - With the goal of increasing the safety of plasma used in the manufacture of therapeutic products, Immuno and its subsidiary Community Bio-Resources (now a division of Baxter Healthcare Corporation), have developed a comprehensive plasma quality programme. This programme includes four main safety initiatives: a plasma centre location/appearance programme, a Qualified Donor programme, an Inventory Hold, and the PCR testing of plasma pools. Many of these initiatives have been adopted in part by the plasma collection and fractionation industry. Using a statistical model that takes into consideration the unique donation characteristics of remunerated plasma donors, combined with 1998 CBR virus reactive rates, an estimated residual likelihood of an undetected donation entering a plasma pool was determined. These estimates, for each million donations, were 0, 1.64, and 4.68 donations for HIV, HBV, and HCV, respectively, and were far below those previously reported for remunerated or volunteer donations. These estimates were confirmed by subsequent PCR testing, which allowed for the additional removal of positive units before manufacture. The low virus load of this plasma supply, combined with increasingly effective virus removal and inactivation procedures, has resulted in the safest ever supply of plasma derivatives. PMID- 10794088 TI - Non-enveloped viruses transmitted by blood and blood products. AB - Since the advent of solvent detergent (S-D) treatment for inactivation of enveloped viruses, there has been no transmission of human immunodeficiency virus, hepatitis B virus, or hepatitis C virus by treated blood products. However, shortly after the introduction of S-D treatment, transmission of hepatitis A with S-D treated factor concentrates was reported in Germany, Italy, Ireland, the United States and South Africa, and this raised awareness of the potential for blood transmission of non-enveloped viruses in general. This report summarizes the physical and epidemiological features of three non-enveloped viruses, hepatitis A virus, parvovirus B19, and the recently identified TT virus, and their transmission by blood and blood products. PMID- 10794087 TI - Risk of hepatitis and retroviral infections among blood donors and introduction of nucleic acid testing (NAT). AB - Blood donors represent a carefully selected population of individuals. In particular, they have reduced prevalence and incidence rates for hepatitis and retroviral markers when compared to the general population. Serological testing of all donations further decreases the risk of collecting an infectious blood unit to negligible levels. Nevertheless, a significant fraction of this residual risk will be eliminated by the introduction of nucleic acid testing for viral genomic material in donor samples. This testing, which has already reached the initial phases of implementation, will be performed on pooled samples, at least in the first instance. PMID- 10794090 TI - The risk of blood-borne Creutzfeldt--Jakob disease. AB - With the increasing ability of modern medicine to transfer life-giving tissues and tissue products from one individual to another comes a parallel, often life taking consequence of the unsuspected transfer of infectious passengers. Creutzfeldt-Jakob disease (CJD) has been transmitted by contaminated stereotactic EEG electrodes, neurosurgical instruments, dura mater allografts, and cadaveric pituitary hormone therapy. Widespread concern has been voiced about the possible risks of transmitting CJD through the administration of blood or plasma products, although no such case has so far been identified by epidemiological studies of at risk populations. Recently completed experiments using hamsters and mice showed decreasing levels of infectivity in buffy coat, plasma, cryoprecipitate (the source of anti-haemophilic factor), and fraction I + II + III (the source of immune globulin). Preliminary results from further rodent experiments indicate that little or no infectivity is detectable in plasma during the pre-clinical phase of infection, that plasma infectivity is not reduced by either leukodepletion filtration or high speed centrifugation, and that about 5-10 times more infectivity is needed to transmit disease by the intravenous than intracerebral route. Altogether, the data from these rodent studies, especially when considered together with results from similar studies underway in primates, should provide a rational foundation for policy decisions by government agencies, plasma product manufacturers, and blood banking communities. PMID- 10794091 TI - Residual risk of transfusion transmitted viral infections among seronegative donors: application of the incidence/window period model. PMID- 10794092 TI - Comparative methods of viral gene amplification. PMID- 10794093 TI - Experience with PCR screening. AB - The COBAS AmpliScreen Hepatitis C (HCV) Test, Version 2.0, which is designed for screening pools composed of samples from individual units of blood or plasma, employs a MultiPrep sample processing procedure that simultaneously extracts and concentrates HCV, HIV-1 and Hepatitis B virus particles from plasma. An HCV Internal Control (IC) RNA serves as an extraction and amplification control for each independently processed sample. Processed samples are amplified by RT-PCR using HCV-specific complementary primers and detected by hybridization of the amplified products to HCV- and IC-specific oligonucleotide probes. The analytical sensitivity of the test is 25 International Units (IU) of HCV per mL of pooled plasma; all HCV genotypes are detected with similar efficiency. The test detected HCV RNA 23 to 32 days prior to anti-HCV antibody seroconversion for four of the five seroconversion panels tested. The test had sufficient sensitivity to reproducibly detect a single infected unit containing 2.4 x 10(3) copies of HCV per mL in a pool with 23 uninfected units. COBAS AmpliScreen tests for HIV-1 and HBV now being validated by Roche Molecular Systems also incorporate the MultiPrep specimen processing method, thereby making it possible to use a single processed specimen to screen for all three viruses. PMID- 10794095 TI - Experiences with genetic screening for established and new agents. AB - In England the introduction of PCR for HCV RNA in minipools has been a nationally coordinated project. The current programme is designed to ensure that all frozen components will have been tested for HCV RNA on pools of 96 samples before their release. To meet EEC requirements, the original planning was aimed at the PCR testing of minipools of 480 samples before making final pools of plasma for fractionation. However, with the sourcing of plasma for fractionation from the USA as a precaution against possible risk from new variant CJD, the project was modified to allow release of HCV PCR tested components. PMID- 10794094 TI - Polymerase chain reaction in detecting hepatitis C virus among blood donors. PMID- 10794096 TI - Implementation of HCV-NAT testing in The Netherlands. PMID- 10794098 TI - Inactivation of viruses, bacteria, protozoa and leukocytes in platelet and red cell concentrates. AB - Despite the increased safety of blood achieved through continued improvements in donor testing, concern remains about the safety of blood components. Transfusion of cellular components has been implicated in transmission of viral, bacterial, and protozoan diseases [1]. While it is commonly recognized that hepatitis B virus, hepatitis C virus, cytomegalovirus, and the retroviruses, such as human immunodeficiency virus and the human lymphotrophic viruses can be transmitted through cellular components, other pathogens are emerging as potentially significant transfusion-associated infectious agents. For example, transmission of protozoan infections due to trypanosomes [2-4] and babesia [5] have been reported. In addition to viral and protozoal infectious agents, bacterial contamination of platelet and red cell concentrates continues to be reported [6, 7] and may be an under-reported transfusion complication [8]. More importantly, new infectious agents, such as HIV, may periodically enter the donor population before they can be identified. During the past decade a number of methods to inactivate infectious pathogens in blood components have been investigated. This technology is now in the clinical trial phase. PMID- 10794097 TI - Large scale PCR screening of pooled plasma samples for HIV-1 and HCV. PMID- 10794099 TI - The use of dimethylmethylene blue for virus photoinactivation of red cell suspensions. AB - Phenothiazine dyes and light have been known to have virucidal properties for over seventy years. This review will describe recent progress in the use of one phenothiazine dye, dimethyl-methylene blue, for photo-inactivation of a number of RNA and DNA viruses in red cell suspensions under conditions that minimally affect red cell in vitro properties during 42-day 1-6 degrees C storage. Dimethylmethylene blue has a higher affinity for nucleic acid than the closely related phenothiazine, methylene blue. Virus photoinactivation appears to be mediated by singlet oxygen. The kinetics of photoinactivation depends on the virus studied, but for a given virus, is similar for both intracellular and extracellular forms. The similarity for inactivation of intracellular and extracellular virus suggests that a common target, such as nucleic acid, is involved. Finally, lymphocytes, which can harbour transfusion-associated viruses and can mediate transfusion-associated-graft-versus host disease, are sensitive to dimethylmethylene blue photoinactivation under virucidal conditions. PMID- 10794100 TI - Inactivation of viruses by aziridines. AB - Ethyleneimine (EI) and N-acetylethyleneimine (AEI) have been shown to inactivate viruses belonging to most of the families described by the International Committee for the Taxonomy of Viruses. The mechanism by which they inactivate the viruses has not been established. In this paper, experiments with foot-and-mouth disease virus (FMDV) and poliovirus are described which indicate that the inactivating lesion is on the RNA. PMID- 10794101 TI - Properties of cyanovirin-N (CV-N): inactivation of HIV-1 by sessile cyanovirin-N (sCV-N). AB - Cyanovirin-N (CV-N) is a novel anti-HIV protein isolated and characterized from a cyanobacterium Nostoc ellipsosporum. CV-N protein is a single 101 amino acid chain containing two intrachain disulphide bonds and considerable internal sequence duplication, but no significant homology to previously described proteins or to the transcription products of known nucleotide sequences. In solution, CV-N exists largely as a beta-sheet protein with internal two-fold pseudosymmetry. CV-N irreversibly inactivates diverse laboratory strains, primary isolates and clades of HIV-1, as well as strains of HIV-2 and simian immunodeficiency virus (SIV). CV-N binds with extremely high affinity to highly conserved binding site(s) on the viral envelope glycoprotein gp120, preventing virus-to-cell fusion, viral entry and infection of cells. The CV-N binding site appears to overlap, but is not identical with, the unique carbohydrate-dependent epitope 2G12, and may lie predominantly within an immunologically "silent" region of gp120. CV-N is undergoing preclinical development for topical anti-HIV prophylactic (e.g., microbicidal) applications to prevent sexual transmission of HIV. Since CV-N may be immunogenic in humans, methods for using CV-N for ex vivo inactivation of HIV in blood, plasma, or putative vaccines preferably would allow for its exclusion from biologicals for parenteral use. To explore this concept we biotinylated CV-N (bCV-N) and coupled it to streptavidin coated magnetic beads to provide a product which we termed sessile CV-N (sCV-N). When reacted with a laboratory strain and a primary isolate of HIV- 1, the sCV-N completely inactivated 100 TCID50 of the virus. However RT-PCR of the viral extracts indicated that only a fraction of the virus was removed by the sCV-N, leaving behind a relatively larger fraction of non-infectious virus in the supernatant which we designated as replication incompetent virions (RIV). It would be worthwhile investigating the role of RIV as a putative HIV vaccine. PMID- 10794102 TI - Photochemical decontamination of red blood cell concentrates with the silicon phthalocyanine PC 4 and red light. AB - Various approaches are being developed for virus inactivation of red blood cell concentrates (RBCC) in order to increase the safety of the blood supply. We have been studying the silicon phthalocyanine Pc 4 for this purpose, a photosensitizer activated with red light. Pc 4 targets the envelope of pathogenic viruses such as HIV. To protect RBC during the process two main approaches are used: (i) inclusion of quenchers of reactive oxygen species produced during the treatment. Tocopherol succinate was found to be most effective for this purpose; (ii) formulation of Pc 4, a lipophilic compound, in liposomes that reduce its binding to RBC but not to viruses. As a light source we used a light emitting diode array emitting at 670-680 nm. An efficient mixing device ensures homogenous light exposure during treatment of intact RBCC. Treatment of 50 ml RBCC with 5 microM Pc 4 and 18 J/cm(2) light results in the inactivation of > or = 5.5 log(10) HIV, > or = 6.3 log(10), VSV and > or = 5 log(10) of PRV and BVDV. The relative sensitivities of these viruses based on the slope of virus kill versus light dose are 1.0, 1.25, 1.5 and 1.9 for HIV, VSV, PRV and BVDV, respectively. To achieve the same level of virus inactivation in 350 ml RBCC, the light dose needed is 40 J/cm(2). HIV actively replicating in CEM cells is as sensitive as cell-free and HIV in latently infected cells is 3-4 times more sensitive. Parasites that can be transmitted by blood transfusion (P. falciparum and T. cruzi) are even more sensitive than viruses. Following treatment, RBCC can be stored for 28 days at 4 degrees C with haemolysis below 1%. Previous studies under less favourable conditions showed that baboon RBC circulated with an acceptable 24 hr recovery and half-life. Genetic toxicological studies of Pc 4 with or without light exposure (mutagenicity in bacteria, mammalian cells in vitro and clastogenicity in vivo) were negative. We conclude that a process using Pc 4 and red light can potentially reduce the risk of transmitting pathogens in RBCC. PMID- 10794103 TI - Membrane filtration for virus removal. AB - Incorporation of adequate clearance methodologies for viral safety assurance constitutes an integral part of the manufacturing process for plasma-derived biologicals. Filtration, being non-invasive, non-destructive and a <> mechanism for virus removal is often the method of choice. This paper discusses the virus filter performance characteristics and other process-related issues specifically applying to the Pall Ultipor VF filters (Grade DV50 and Grade DV20), which are operated in the direct flow filtration (DFF) mode and the Pall Filtron Omega VR filters (the Omega 300 K and Omega 100 K), which are operated in the tangential flow filtration (TFF) mode. PMID- 10794105 TI - Reducing the risk of bacterial contamination of cellular blood components. AB - Transfusion-associated septic reactions occurring during or following the transfusion of cellular blood components was one of the earliest recognised complications of allogeneic blood transfusions. The presence of bacteria in cellular blood products thus has been a problem for many decades and currently it is the most common microbiological cause of transfusion-associated morbidity and mortality. Transfusion-associated septic reactions due to contaminated platelet concentrates appear to be much more common than those due to red cell concentrates. The prevalence of contaminated cellular blood products (red cells and platelets) is approximately 1 in 2,000. However, the transfusion to a recipient of a contaminated blood product may not be associated with morbidity, because many contaminated blood product units contain only few bacteria and such transfusions may be innocuous to the recipient. In other instances, contaminated blood product units may contain large numbers of virulent bacteria and endotoxins, and their transfusion may be associated with significant morbidity and may even be lethal to the recipient. The prevalence of severe episodes of transfusion-associated sepsis has not been clearly established, but is probably of the order of 1 in 50,000 for platelet units and 1 in 500,000 for red blood cell units transfused. As a result of the increased recognition that such transfusion-associated septic episodes can occur, a variety of measures have been proposed to try to prevent and/or control the rate of contamination of blood products. PMID- 10794104 TI - Safety and utility of blood substitutes. PMID- 10794106 TI - Transfusion safety in developing countries and the Indian scenario. AB - The AIDS pandemic has brought into focus the importance of safe blood transfusion. The management of an effective transfusion service is an expensive endeavour even in the most developed countries, and is therefore a monumental challenge for developing countries with limited budgets and other priorities. HIV prevalence in the Indian population has shown a steady rise from 0.5% in 1990 to 1.2% in 1997 with the highest prevalence in cities. When the HIV infection was discovered in India in 1986, the health authorities set up the National AIDS Control Organisation (NACO) with a primary focus on ensuring a safe blood supply. NACO was funded by the World Bank and technically supported by WHO. The supreme Court of India has also taken up the issue of blood safety by banning paid donations by the end of 1997 and established the autonomous National Blood Transfusion Council and the State Transfusion Councils. The Drugs Controller of India and State F.D.A. have issued licences to all blood banks to streamline them after all requirements are met. However, there are a number of blood banks which are operating without licences. While India collects three million units of blood, barely 10% is available as blood components, and only a percentage of the blood is being screened for infectious markers. Nevertheless, there is a general recognition that an improved transfusion service is required in India. PMID- 10794107 TI - Worldwide perspective in transfusion safety: Europe. PMID- 10794109 TI - Achievement of universal blood safety: introduction to the International Consortium for Blood Safety (ICBS). PMID- 10794110 TI - The FDA approach to standardization and quality assurance of nucleic acid tests used to screen blood and plasma donations. PMID- 10794108 TI - Cost-effectiveness of new blood safety technologies. AB - Decision analysis techniques can be used to project the health benefit obtained by commitment of a given amount of resources. Such analyses can help physicians choose approaches offering the greatest benefit with the least risk, and can help health planners direct resources to where they will provide the greatest benefit. As the risks of allogeneic transfusion continue to decline, the yield of additional safety improvements necessarily diminishes, making their cost effectiveness astronomically poor. While society may choose to direct our efforts in this direction, the value obtained by focussing attention on these areas of minimal yield should be clarified. PMID- 10794111 TI - Cost and public perception. PMID- 10794113 TI - Studies on the psychosomatic functioning of ill-health according to Eastern and Western medicine. 4. The verification of possible links between ill-health, lifestyle illness and stress-related disease. AB - In our previous reports on the same general topic, we exploited research in Oriental psychosomatic medicine to obtain diagnoses through visual observation of the sublingual vein (useful for the early detection of vital energy stagnation and blood stasis) from a computerized color analysis of the tongue proper, its coating, and the sublingual vein. This led us to develop the concept of the anxiety-affinitive constitution, based on unbalanced qi, blood, and body fluid in ill-health as a causative factor for stress-related diseases and lifestyle illnesses. As a development of this research, the present report examines the verification of possible links of ill-health, lifestyle illness, and stress related disease through the diagnosis and treatment of functional subclinical psychosomatic disorders detected as a clinical expression of the anxiety affinitive constitution. First, a diagnosis of functional subclinical psychosomatic disorders (ill-health) in 197 medical school students, including 156 first-year students at medical and nursing schools made it clear that the trait anxiety forming the core of the anxiety-affinitive constitution is linked to Dr. Lester Breslow's seven good health habits, and the manner of respiration. Second, it was revealed during the treatment of functional subclinical psychosomatic disorders in eight medical students that kampo medication and relaxation training (RT) produce lowered scores for STAI trait anxiety, and transform respiration from thoracic pattern to a balanced thoracic and abdominal respiratory pattern. This was particularly true for RT. Finally, we concluded that a high score for trait anxiety correlates to the formation of inappropriate health habits, and the habituation of inadequate respiration (thoracic pattern). Therefore, lifestyle illness or stress-related diseases will develop unless an anxiety-affinitive constitution is improved with kampo medication and/or RT. PMID- 10794112 TI - Seeds induced to germinate rapidly by mentally projected 'qi energy' are apparently genetically altered. AB - Mentally controlled qi energy can induce crop seeds to sprout and root for several cm within about 20 min. The RAPD method was used to compare treated groups of wheat and pea seeds and their controls using 11 selected primers. Seven primers amplified polymorphisms in wheat seeds and 5 in pea seeds. It was thought preliminarily that qi energy changed the structure of a germination-correlated gene site speeding up expression and advancing it in time. PMID- 10794114 TI - Effect of acupuncture on pain management in patients before and after lumbar disc protrusion surgery--a randomized control study. AB - Management of acute and chronic low back and leg pain often includes the use of acupuncture. The effectiveness of this form of therapy is dependent upon compliance, which in turn is dependent on availability, response, treatment of proper acupoints, and the placebo effect. We hypothesized that classical acupuncture would be more effective than placebo acupuncture. One hundred and thirty-two patients with acute and chronic low back and leg pain were examined before and after surgery for lumbar disc protrusion. Diagnosis was based on CT and MRT findings. Patients received acupuncture drug-free throughout the study period. The visual analogue scale was used to assess pain intensity before and after (i.e. 30 min. 60 min. 2 h and 6 h) acupuncture. Classical acupuncture resulted in a significant reduction in pain that become increasingly stronger during the 6h study period. Placebo acupuncture lead to same early pain relief that did not reach statistic significant and then declined thereafter. PMID- 10794115 TI - The effect of acupuncture on uterine contraction induced by oxytocin. AB - Preterm labor (PTL) is one of the main causes of fetal mortality and morbidity in obstetrical medicine. Current methods of treatment are not very effective and often have significant side effects. For this reason new methods of preventing PTL are currently being sought. In Western medicine the newest development is oxytocin antagonists. In Oriental medicine acupuncture and moxibustion are being utilized for the purpose of stopping PTL. The goals of this study were to determine if acupuncture in pregnant rats can suppress oxytocin induced uterine contractions and to compare these results with those inhibited by an oxytocin antagonist. Uterine contractions were induced by continuous infusion of exogenous oxytocin. The first fetus in one uterine horn near the ovarian end was removed and distilled water-filled catheter was inserted into that vacated amniotic sac to measure uterine contractions as intrauterine pressure changes. Two acupoints of Ho-Ku (LI-4) and San-Yin-Chiao (Sp-6) were selected for acupuncture and Kuan Yuan (Co-4) was used for moxibustion. The oxytocin-induced uterine contractions were significantly suppressed by acupuncture on the LI-4 (p < 0.05), but not by Sp-6. Stimulation of Co-4 by moxibustion had no significant (p > 0.05) tocolytic effect. The administration of oxytocin antagonist eliminated all the uterine contractions induced by oxytocin. The application of acupuncture to re-stimulate the activity that was suppressed by the oxytocin antagonist did not produce any positive results. However, prostaglandins did cause the uterus to contract. In conclusion, acupuncture on LI-4 was found to suppress uterine contractions induced by oxytocin in the pregnant rat. If acupuncture is similarly effective in counteracting the effects of oxytocin in women, then this may an alternative medical treatment for women in preterm labor. PMID- 10794116 TI - Effect of acupuncture at Hsien-Ku (St-43) on the pulse spectrum and a discussion of the evidence for the frequency structure of Chinese medicine. AB - We investigated the pulse spectrum variation of the human radial artery when Hsien- Ku (St 43), an acupoint on the stomach meridian, was needled and compared the results with the acupuncture effects of two other acupoints, Tsu-San-Li (St 36) and Tai-Shih (K-3), reported previously. For Hsien-Ku, the harmonic proportions were redistributed: the second harmonic (C2) decreased, C3, C5, C6, C7, C8 and C9 increased, C3, C6 and C9 became the relative peaks to their neighboring harmonics and C2, C4 became the relative minimums. The phase angles of the 2nd harmonic (P2) and 5th harmonic (P5) decreased, propagating faster. These effects were similar to that of Tsu-San-Li which is also on the stomach meridian. A totally different pattern was found for Tai-Shih on the kidney meridian. These results strengthen the theory that a meridian can be classified according to its effects on the pulse spectrum, and that all the meridian related effects such as those caused by acupuncture or meridian specific herbs are frequency specific. PMID- 10794117 TI - Inhibition of human smooth muscle cell proliferation by gamigeonsim-tang through the transcriptional regulation of cell cycle-controlling genes. AB - The effects of gamigeonsim-tang (GGT) on cellular proliferation and expression of cell cycle-related genes were investigated in human smooth muscle cell HISM. HISM cells were treated with an aqueous extract of GGT. Cellular proliferation was investigated by an immunocytometric analysis of PCNA expression and a flow cytometric analysis of the cell cycle progression. Reduced expression of PCNA and a significant accumulation of G1 phase cells were observed following treatment, indicating that GGT inhibits cellular proliferation of human smooth muscle cells. To explore whether GGT affects the transcription of cell cycle-regulating genes, we evaluated mRNA expression of p53, p21Waf1 PCNA, Cyclin D1, Cdc2, Histone H3, c Myc, and c-Fos using a quantitative RT-PCR analysis. While increased expressions of two negative cell cycle regulators, p53 and p21Waf1 were found, reduced expressions of cell cycle stimulators, PCNA, c-Fos, and c-Myc, were identified following treatment. Taken together, our study demonstrates that GGT inhibits cellular proliferation of human smooth muscle cell through the up- and down regulation of growth-inhibiting and growth-promoting genes, respectively. PMID- 10794118 TI - Inhibition of mast cell-mediated anaphylaxis by sochungryong-tang. AB - According to traditional Asian philosophy, sochungryong-tang (S-Tang) is a prescription for treating exterior syndrome. In this study, we investigated the effect of S-Tang on mast cell-mediated anaphylaxis. S-Tang completely inhibited compound 48/80-induced systemic anaphylactic shock at a dose of 100 mg/kg. When S Tang was given as pretreatment at concentrations ranging from 1 to 1000 mg/kg, the serum histamine levels induced by compound 48/80 were reduced in a dose dependent manner. S-Tang inhibited the local anaphylaxis activated by anti dinitrophenyl (DNP) IgE antibody, and also inhibited the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. These results indicate that S-Tang may contain substances with actions that inhibit mast cell degranulation. PMID- 10794119 TI - Anti-aging effects of the traditional Chinese medicine bu-zhong-yi-qi-tang in mice. AB - Among the "alternative medicines," which may ably supplement modern Western medicine in the treatment of certain diseases, the holistic approach and mild nature of the majority of Traditional Chinese Medicine (TCM) may make it particularly suitable for the treatment of diseases associated with old age, as the general health of elderly patients is already compromised. The TCM formulation of bu-zhong-yi-qi-tang (B.Z.Y.Q.T.), prescribed mainly for the improvement of circulation and in particular that to the gastroenteric regions, may have anti-aging effects. In the present study, possible anti-aging effects of B.Z.Y.Q.T. were studied using normal (ICR) mice and the Dull, P/8 and R/1 strains of the Senescence Accelerated Mouse (S.A.M.). Following repeated oral administrations of B.Z.Y.Q.T. at 250 and 500 mg/kg the test mice were assessed for (1) endurance (2) learning and memory (3) neuromuscular coordination and (4) changes in the levels of monoamines in the brain. The results indicated that B.Z.Y.Q.T. improved endurance in all strains in a dose-dependent manner. At the higher dose of 500 mg/kg, it improved memory in the R/1 and P/8 S.A.M. mice. In prolonged rota-rod tests, which assessed both motor coordination and endurance, B.Z.Y.Q.T. significantly improved performance in the P/8 S.A.M. mice. Elevated dopamine and noradrenaline were observed in cortical tissues of the S.A.M./Dull and ICR mice respectively with the high dose of 500 mg/Kg, B.Z.Y.Q.T. Taken together, the results indicated that B.Z.Y.Q.T. appeared to exert anti-aging effects in mice and elevation in certain monoamines in brain cortical tissues. How and whether the monoamines changes after B.Z.Y.Q.T. treatment might be related to the behavioral effects await further investigation. PMID- 10794120 TI - Antioxidant and hepatoprotective effects of Anoectochilus formosanus and Gynostemma pentaphyllum. AB - Anoectochilus formosanus Hay. and Gynostemma pentaphyllum Makino are popular folk medicines that have been used for treating hepatitis, hypertension and cancer in Taiwan. Our previous studies showed that these crude drugs exert antiinflammatory activity and hepatoprotective activity against CC14-induced liver damage. In this study, the antioxidant effect of these crude drugs and their hepatoprotective activity on acetaminophen-induced liver injury in rat was evaluated. Our results suggest that A. formosanus and G. pentaphyllum do have antioxidant effects. On acetaminophen-intoxicated model, the increased levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) by acetaminophen administration were reduced by treatment with these two herbs. In histological observation, gross necrosis in the centribular area, sinusoidal congestion, infiltration of the lymphocytes and Kupffer cells around the hepatic central vein, and loss of cell boundaries and ballooning degeneration were reduced with herbal treatment. However, the effect of A. formosanus and G. pentaphyllum is biphasic. Methanol extract (100 and 300 mg/kg) and water extract (300 and 500 mg/kg) of A formosanus and water extract (100, 300 and 500 mg/kg) of G. pentaphyllum enhanced the recovery of liver injury while treatment with 500 mg/kg of A. formosanus methanol extract resulted in serious hepatic injury. PMID- 10794122 TI - The hepatoprotective effects of Solanum alatum Moench. on acetaminophen-induced hepatotoxicity in mice. AB - Solanum alatum Moench. has been shown to have a protective effect against carbon tetrachloride (CCl4)-induced liver injury. Solanum alatum treatment (100 mg/kg, p.o.) decreased the elevation of serum alanine aminotransferase (ALT; GPT) and aspartate aminotransferase (AST; GOT) induced by acetaminophen (paracetamol) (600 mg/kg, i.p.) administration. It also decreased the extent of visible necrosis in liver tissue. In addition, Solanum alatum treatment restored hepatic glutathione (GSH) depletion induced by acetaminophen (600 mg/kg, i.p.) administration. Microsomal enzyme levels such as P-450, reductase, and aniline hydroxylation enzyme were also restored to normal levels after Solanum alatum administration. The hepatoprotective mechanism may function through direct binding with acetaminophen toxic metabolites, decreasing the attraction of acetaminophen metabolites for other cellular GSH or thiol protein. Additionally, Solanum alatum treatment increased the concentration of hepatic GSH and maintained a high level activity of GSTase, which led to acceleration of the excretion of toxic acetaminophen metabolites. PMID- 10794121 TI - The effects of traditional tonics on fatigue in mice differ from those of the antidepressant imipramine: a pharmacological and behavioral study. AB - The present studies were undertaken to investigate the differences between the antidepressant drug, imipramine, and liquid nutritive and tonic drugs (NTDs) that consist of Ginseng radix, Epimedii herba, Holen and an additional eight to twelve crude drugs. After preloading forced swimming, the NTD (applied orally, 0.1 ml/10 g) significantly increased the duration time of swimming and decreased the duration time of immobility, while the administration of imipramine (5, 10 and 20 mg/kg, i.p.) under the same conditions and after the same treatment did not produce these positive effects. After pretreatment with 100 mg/kg tetrabenazine, the NTDs also elicited both the increased locomotor activity and the decreased duration time of immobility. The behavioral effect was similar to treatment with imipramine. The NTDs showed a long lasting effect on swimming behavior in the forced swimming test for 15 min, indicating a prolonged efficacy, not like the short effect of imipramine. The present results indicate that the effect of NTDs on fatigued subjects is different from that of imipramine, probably due to involvement of another factor in addition to the antidepressant effect. PMID- 10794124 TI - A study on the toxicology of Acacia nilotica. AB - The potential toxicity of Acacia nilotica was investigated in rats maintained on 2% and 8% acacia diet for 2 and 4 weeks. A significant reduction in body weight in all acacia-fed groups and a significant decrease in the levels of hemoglobin, serum total protein and total cholesterol in animals fed 8% acacia diet for up to 4 weeks were observed. These effects were, however, reversed one week after treatment termination. No significant changes in serum parameters of hepatic and renal functions, fasting glucose and triglycerides were observed. Further, no deaths among treated animals and no significant histopathological changes in liver sections were noted. It is concluded that A. nilotica, at 2% and 8% levels, has a low toxicity potential. PMID- 10794123 TI - Hepatoprotection by "dangqui-long-hwei-wan" in male mice. AB - Dangqui-long-hwei-wan (D.L.H.W.) is a traditional Chinese prescription for treatment of hepatitis. The hepatoprotective effects of D.L.H.W. and its constituents were investigated on carbon tetrachloride-induced liver damage mice. The hepatoprotective effect is more prominent for aqueous extracts of the complete formula of D.L.H.W., especially the one cited in the Chinese medical book Hsuan-Ming-Lun (H.M.L.). Our results are in accordance with those described in the Chinese medical literature and the indications suggested in clinical treatment (Wang, 1982). The results further indicate that Scutellariae Radix plays an important role in the hepatoprotective activity. Moschus could be omitted from D.L.H.W. with no significant influence on its effect. The underlying mechanism for the hepatoprotective effect of D.L.H.W. possibly results from the inhibition of the formation of *CCl3 and the enhancement of immunity of hepatitis carrying patients. PMID- 10794125 TI - The choice between Chinese medicine and Western medicine practitioners by Hong Kong adolescents. AB - A survey of 3,355 Hong Kong adolescent students showed that the prevalence of TCM clinic consultation in a three-month period was 8.6%. Among students who sought doctor consultation in a three-month period, 17.8% consulted a TCM doctor. Students who sought doctor consultation for fall-related injuries were more likely to have consulted TCM doctors only, while those for skin disease were more likely to have consulted both TCM and western doctors. Perceived cost, efficacy, trust of western doctors and socioeconomic factors were not associated with choice of doctor type. Our findings are different from those on Hong Kong adults' use of health care services as reported in other studies. PMID- 10794126 TI - Preventing and resolving post-laparotomy intestinal obstruction: an effective shiatsu method. PMID- 10794127 TI - Anesthetic and analgesic practices in Avicenna's Canon of Medicine. AB - Anesthetic and analgesic practices during the Middle Ages and the Renaissance are presented from a translation of the relevant sections of the Canon of Medicine by Avicenna (980-1037) one of the most widely read and authoritative textbooks of the period. Avicenna described tracheotomy, oropharyngeal intubation and a method for clearing upper airway secretions for the treatment of stridor and respiratory distress. He also identified certain plants with pharmacological action such as mandragora or nightshade, opium and henbane and gives various recipes for inducing both anesthesia and analgesia before surgery. PMID- 10794129 TI - Taurine release modified by GABAergic agents in hippocampal slices from adult and developing mice. AB - In order to characterize the possible regulation of taurine release by GABAergic terminals, the effects of several agonists and antagonists of GABA receptors on the basal and K+-stimulated release of [3H]taurine were investigated in hippocampal slices from adult (3-month-old) and developing (7-day-old) mice using a superfusion system. Taurine release was concentration-dependently potentiated by GABA, which effect was reduced by phaclofen, saclofen and (1,2,5,6 tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA) at both ages, suggesting regulation by both GABA(B) and GABA(C) receptors. The involvement of GABA(A) receptors could not be excluded since the antagonist bicuculline was able to affect both basal and K+-evoked taurine release. Furthermore, several GABA(B) receptor effectors were able to inhibit K+-stimulated taurine release in the adults, while the GABA(C) receptor agonists trans-4-aminocrotonic acid (TACA) and cis-4-aminocrotonic acid (CACA) potentiated this release. The potentiation of taurine release by agents acting on the three types of GABA receptors in both adult and developing hippocampus further indicates the involvement of transporters operating in an outward direction. This inference is corroborated by the moderate but significant inhibition of taurine uptake by the same compounds. PMID- 10794128 TI - Regulation of growth hormone (GH) secretion by different glutamate receptor subtypes in the rat. AB - It has been firmly established that excitatory amino acids (EAAs), such as glutamate, are pivotal elements in the hypothalamic circuitry involved in the control of pituitary function. The actions of EAAs are mediated by different postsynaptic receptor subtypes, which include N-methyl D-aspartate (NMDA), kainate (KA), 2-amino-3-hydroxy-5 methyl-4-isoxazol propionic acid (AMPA) and metabotropic receptors. In this review, we summarize our experimental work on the role of EAA neurotransmission in the control of GH secretion in the rat. Detailed characterization of the effects of agonists and antagonists of glutamate receptors on GH release revealed that activation of NMDA, KA and AMPA receptors at different age-points resulted in clear-cut stimulation of GH secretion, although age- and sex-dependent differences were detected in the pattern of response to the different agonists. This stimulatory action was proven nitric oxide (NO)-dependent and not exerted at the pituitary level. In addition, evaluation of the role of hypothalamic GH-releasing hormone (GHRH) in the stimulatory action of NMDA by means of immunoneutralization of endogenous GHRH or destruction of GHRH producing neurons suggested the involvement of signals other than GHRH in this response. Further, evidence was obtained on the modulation of the EAA system by gonadal factors, and on the physiological relevance of EAA pathways in the regulation of pulsatile GH release. In conclusion, our data using the rat as animal model provide evidence for a pivotal role of glutamate pathways in the regulation of GH secretion throughout the life-span. PMID- 10794130 TI - Cerebral organic acid disorders induce neuronal damage via excitotoxic organic acids in vitro. AB - Glutaryl-CoA dehydrogenase deficiency (GDD), which is one of the most frequent organic acid disorders, is characterized by a specific age- and regional dependent neuropathology. We hypothesized that the distinct brain damage in GDD could be caused by the main pathologic metabolites, the organic acids glutaric (GA) and 3-hydroxyglutaric (3-OH-GA) acids, through an excitotoxic sequence. Therefore, we investigated the effects of 3-OH-GA and GA on primary neuronal cultures from chick embryonic telencephalons. Here we report that 3-OH-GA and GA decreased cell viability concentration- and time-dependently, which could be only totally prevented by preincubation with MK-801, ifenprodil and NR2B antibodies. Furthermore, cell viability decreased in parallel with the increasing expression of NR2B subunit on cultured neurons from 2nd to 6th DIV. We conclude that GA and 3-OH-GA act as excitotoxic organic acids (EOA) specifically through NR1/NR2B and that the extent of induced neurotoxicity is dependent on NR1/NR2B expression during maturation. PMID- 10794131 TI - Synthesis and biological activity of canavanine hydrazide derivatives. AB - The canavanine derivatives L-canavanine hydrazide (CH), L-canavanine-bis-(2 chloroethyl)hydrazide (CBCH) and L-canavanine phenylhydrazide (CPH) were synthesized and evaluated for biological activity in microorganisms, plants and tumor cells using canavanine as a positive control. (1) In microbial systems, the compounds exerted activity, as assessed in 14 bacterial strains. The effect of canavanine was easily removed by equimolar concentrations of arginine or ornithine, while the effect of CBCH or CPH was abolished by 10-fold excess of arginine or 10- to 100-fold excess of ornithine. (2) In plants, the activity of CH and CBCH were relatively low, whereas the inhibitory potential of CPH was comparable or even superior to that of canavanine, resulting at 1 mM concentration in a nearly complete block of tomato cell growth, and reducing by up to 80% the length of radicles of cress, amaranth, cabbage and pumpkin. (3) In pumpkin seeds, CPH or canavanine induced the synthesis of four small heat shock proteins of hsp-17 family in the pH range of 6 to 7.5. The proteins exhibited in both cases a similar profile, but differed in the timing of their expression and/or accumulation. With canavanine, the highest hsp-17 expression was found after 48 h of drug treatment, while with CPH this maximum was shifted to 24 h. (4) CPH proved to be highly cytotoxic against Friend leukemia cells in culture, exceeding by one order of magnitude the cytotoxicity of canavanine. The effect of canavanine was completely removed in the presence of equimolar amounts of arginine, while a 20-fold excess of arginine failed to abolish the cytotoxicity of CPH. Thus, a proper hydrazide modification of canavanine may lead to a significant increase in its growth-inhibitory activity and to a change in the mode of action of the parent compound. PMID- 10794132 TI - Identification of an oxidation product of aminoethylcysteine ketimine dimer. AB - In continuation of our previous work dedicated to the detection of the oxidation products of aminoethylcysteine ketimine dimer by oxygen reactive species, we give here data for the identification of the alpha, beta unsaturated sulfoxide as the main product of interaction of the dimer with H2O2. Identification has been done on the basis of mass spectrometry and NMR analyses of the product isolated by preparative chromatography. PMID- 10794133 TI - The binding of amino acids to the herbicide 2,4-dichlorophenoxy acetic acid. AB - The interaction of amino acids with the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was studied by charge-transfer chromatography carried out on diatomaceous layers covered with different amount of 2,4-D and the effect of salts on the strength of interaction was elucidated. It was established that Arg, His, Lys, Orn, Phe and Trp binds to 2,4-D, the binding process is of saturation character. Principal component analysis proved that the concentration of 2,4-D exerts the highest impact on the interaction and the effect of salts is of secondary importance. The results suggest that these amino acid residues may account for the binding of 2,4-D to proteins and can play a considerable role in the detoxification processes by forming conjugates with 2,4-D. PMID- 10794134 TI - Stereospecific synthesis of alpha-methylated amino acids. AB - Both 2,5-trans and 2,5-cis disubstituted 2-tert-butyl-5-(indol-3 yl)methylimidazolidin-4-ones were synthesised and their enolates were prepared using LDA. While the enolate of the 2,5-trans disubstituted derivative could not be methylated, the enolate of the cis-2,5-disubstituted derivative was successfully methylated with methyl iodide to a product which on hydrolysis gave enantiomerically pure alpha-methyl-L-tryptophan. PMID- 10794136 TI - N-acyl homoserinelactone-mediated gene regulation in gram-negative bacteria. AB - The view of bacteria as unicellular organisms has strong roots in the tradition of culturing bacteria in liquid media. However, in nature microbial activity is mainly associated with surfaces where bacteria form highly structured and cooperative consortia which are commonly referred to as biofilms. The ability of bacteria to organize structurally and to distribute metabolic activities between the different members of the consortium demands a high degree of coordinated cell cell interaction. Recent work has established that many bacteria employ sophisticated intercellular communication systems that rely on small signal molecules to control the expression of multiple target genes. In Gram-negative bacteria, the most intensively investigated signal molecules are N-acyl-L homoserine lactones (AHLs), which are utilized by the bacteria to monitor their own population densities in a process known as 'quorum sensing'. These density dependent regulatory systems rely on two proteins, an AHL synthase, usually a member of the LuxI family of proteins, and an AHL receptor protein belonging to the LuxR family of transcriptional regulators. At low population densities cells produce a basal level of AHL via the activity of an AHL synthase. As the cell density increases, AHL accumulates in the growth medium. On reaching a critical threshold concentration, the AHL molecule binds to its cognate receptor which in turn leads to the induction/repression of AHL-regulated genes. To date, AHL dependent quorum sensing circuits have been identified in a wide range of gram negative bacteria where they regulate various functions including bioluminescence, plasmid conjugal transfer, biofilm formation, motility, antibiotic biosynthesis, and the production of virulence factors in plant and animal pathogens. Moreover, AHL signal molecules appear to play important roles in the ecology of complex consortia as they allow bacterial populations to interact with each other as well as with their eukaryotic hosts. PMID- 10794135 TI - Derivatives of glutamic acid as new surfactants. AB - Starting from glutamic acid, different types of surfactants have been synthesised by using original trimodular strategies. Monosubstituted zwitterionic amides of glutamic acid obtained with excellent yields show good surface activity. The grafting of a second hydrophobic side-chain leads to bicatenar cationic surfactants or to disubstituted nonionic cyclic compounds. In order to reduce the hydrophobic character of the bicatenar surfactants, a second synthetic method has been developed, allowing the introduction of a polar sugar group into these molecules. The surfactant properties of several of the products have been determined by physico-chemical methods such as surface tension measurements and compression isotherm studies by means of a Langmuir balance. PMID- 10794138 TI - Induction and characterization of Pediococcus acidilactici temperate bacteriophage AB - Mitomycin C was used to induce temperate bacteriophage from three strains of Pediococcus acidilactici. The new bacteriophage, designated pa97, pa40, and pa42, were characterized based on morphology, DNA homology, and major protein profiles. Morphological attributes (small isometric heads with non-contractile tails) place these bacteriophages within the B1 group of the family Siphovirdae. Restriction endonuclease digests suggested that the bacteriophage genomes were linear molecules without cohesive ends, and between 33 and 37 kilobases in length. All three bacteriophages possessed one major protein with an estimated mass of 30 to 35 kilodaltons. Bacteriophage pa42 also contained a second major protein of approximately 47 kilodaltons. DNA-DNA hybridization showed bacteriophages pa40 and pa42 were homologous to each other, but not to pa97, suggesting that Pediococcus acidilactici bacteriophage fall into at least two different species. PMID- 10794137 TI - Influence of lactose-citrate co-metabolism on the differences of growth and energetics in Leuconostoc lactis, Leuconostoc mesenteroides ssp. mesenteroides and Leuconostoc mesenteroides ssp. cremoris AB - The biodiversity of growth and energetics in Leuconostoc sp. has been studied in MRS lactose medium with and without citrate. On lactose alone, Ln. lactis has a growth rate double that of Ln. cremoris and Ln. mesenteroides. The pH is a more critical parameter for Ln. mesenteroides than for Ln. lactis or Ln. cremoris; without pH control Ln. mesenteroides is unable to acidify the medium under pH 4.5, while with pH control and as a consequence of a high Y(ATP) its growth is greater than Ln. lactis and Ln. cremoris. In general, lactose-citrate co metabolism increases the growth rate, the biomass synthesis, the lactose utilisation ratio, and the production of lactate and acetate from lactose catabolism. The combined effect of the pH and the co-metabolism lactose-citrate on the two components of the proton motive force (deltap = deltapsi - ZdeltapH) has been studied using resting-cell experiments. At neutral pH deltap is nearly entirely due to the deltapsi, whereas at acidic pH the deltapH is the major component. On lactose alone, strains have a different aptitude to regulate their intracellular pH value, for Ln. mesenteroides it drastically decreases at acidic pH values (pH, = 5.2 for pH 4), while for Ln. lactis and Ln. cremoris it remains above pH 6. Lactose-citrate co-metabolism allows a better control of pH homeostasis in Ln. mesenteroides, consequently the pHi becomes homogeneous between the three strains studied, for pH 4 it is in an interval of 0.3 pH unit (from pHi = 6.4 to pHi = 6.7). In this metabolic state, and as a consequence of the variation in deltapH, and to some extent in the deltapsi, the difference of deltap between the three strains is restricted to an interval of 20 mV. PMID- 10794141 TI - Serological and molecular size characterization of flagellins of Pseudomonas syringae pathovars and related bacteria AB - Flagella from a total of 118 strains representing mostly pathovars of the phytopathogenic group Pseudomonas syringae, but also P. chlororaphis, P. cichorii, P. corrugata, P. fluorescens, P. fuscovaginae, P. stutzeri, P. viridiflava, as well as related phytopathogenic genera (Burkholderia cepacia and Ralstonia solanacearum) were characterized by immuno-fluorescent staining, SDS PAGE, and immunoblotting. Eighty-six strains of the P. syringae group pathovars, P. cichorii and P. viridiflava were shown to possess flagella of serotypes H1 or H2, composed of a unique flagellin, whose molecular size varied between 31 and 31.5 kDa. Similarities between the P. syringae flagellin and a 31 kDa surface protein involved in pathogenicity are pointed out. The distribution of H1 and H2 antigens in the nine recently described genomospecies of P. syringae-P. viridiflava group suggested that flagellin would represent a phylogenetic marker within the arginin-dihydrolase-negative fluorescent pseudomonads. The characterization of flagellin was proposed as an identification tool at a level situated between genus and species. PMID- 10794139 TI - Genetic organization of the mobilization region of the plasmid pHE1 from Halomonas elongata. AB - The mobilization (mob) region of the non-self transmissible 4.2-kb plasmid pHE1 from the moderately halophilic bacterium Halomonas elongata ATCC 33174 has been identified and characterized. Analysis of the sequence revealed the presence of four open reading frames (mobCABD) which show a complex organization with two of them (mobB and mobD) entirely overlapped by a third (mobA). The deduced proteins appeared to have a high degree of homology to Mob proteins of CoIE1 and closely related plasmids. To assess the functionality of the mob region, the hybrid vector pHS134 was constructed, consisting of the complete plasmid pHEI, the E. coli vector pKS(-) and a streptomycin-resistance gene for positive selection in Halomonas. Vector pHS134 was found to be mobilizable from E. coli to H. elongata assisted by pRK600. Upstream of the mob genes, an oriT region with a putative nick sequence highly homologous to that of CoIE1 plasmids was identified. To our knowledge, this is the first mobilizable plasmid found in moderate halophiles. This property, together with its small size, the availability of its complete sequence, and its broad host range in moderately halophilic strains, makes pHE1 a good candidate for the construction of cloning and expression vectors for these extremophiles. PMID- 10794142 TI - Occurrence of neutral and alkaline cellulases among alkali-tolerant micromycetes AB - 600 different natural strains of alkali-tolerant micromycetes belonging to 4 classes, 38 genera and 119 species were tested on their ability to produce neutral and alkaline cellulases. Frequency of occurrence of neutral and alkaline cellulases producers among the studied strains was estimated. It is concluded that 72% of all tested strains produce cellulases at neutral and alkaline pH values. But the largest number of neutral and alkaline cellulases producers belong to the following classes which are tolerant to extreme environmental factors: namely Hyphomycetes (genera Acremonium, Alternaria, Cylindrocarpon, Fuzarium, Penicillium, Verticillium, Myrothecium, Ulocladium, Gliocladium), Ascomycetes (genus Chaetomium) and Coelomycetes (genera Phoma, Microsphaeropsis, Aposphaeria). PMID- 10794140 TI - Identification of a lipoarabinomannan-like lipoglycan in Gordonia rubropertincta. AB - Lipoarabinomannans are well characterised lipoglycans present in the cell envelopes of mycobacteria and closely related bacteria. To define further the distribution of these lipoglycans we have investigated the mycolic acid containing bacterium Gordonia rubropertincta and, by analysis of carbohydrate composition and antigenic cross-reactivity, demonstrated the presence of a lipoarabinomannan-like lipoglycan. A fraction that appeared to contain higher phosphatidylinositolmannosides was also isolated. PMID- 10794143 TI - Halomonas campisalis sp. nov., a denitrifying, moderately haloalkaliphilic bacterium. AB - The isolation and characterization of a denitrifying bacterium that is both moderately halophilic and alkaliphilic is described. The organism was isolated for use in the development of a bioprocess that could potentially reduce the costs of ion exchange resin regenerant disposal. The process of ion exchange, after resin regeneration, produces a briny, alkaline waste that is difficult and expensive to dispose. The biological removal of nitrate and subsequent reuse of these brines can potentially provide a cost-saving alternative to disposing of this waste product. To achieve our objective, a moderately halophilic, alkaliphilic bacterium was isolated from sediment samples taken from the salt plain of Alkali Lake in Washington State (USA). The haloalkaliphilic bacterium, designated strain 4A, is motile with rod-shaped cells that are 3 to 5 microm long and 1 microm wide. Electron acceptors used include oxygen, nitrate, and nitrite. In addition, it has similar specific nitrate reduction rates and biomass yields as non-halophilic denitrifying bacteria. It is capable of using a variety of electron donors. This organism can grow at NaCl concentrations ranging from 0.2 to 4.5 M with optimum growth occurring at 1.5 M and pH values ranging from 6 to 12 with 9.5 being the optimum pH. The temperature range for growth of strain 4A is 4-50 degrees C with optimal growth occurring at 30 degrees C. The G + C content is 66 mol%. Phylogenetic analyses based upon 16S rDNA gene sequence placed isolate 4A in the genus Halomonas. In addition, DNA-DNA hybridization experiments clearly indicate that it is a unique species. Phenotypic and phylogenetic studies indicate that isolate 4A represents a new species. We propose the name Halomonas campisalis for this species and strain 4A (ATCC 700597) as the type strain. Due to its denitrification ability, broad carbon utilization range and its high salinity and pH tolerance this organism, and similar ones, hold promise for the treatment of saline, alkaline waste. PMID- 10794146 TI - A polyphasic approach towards the identification of strains belonging to Lactobacillus acidophilus and related species. AB - A set of 98 strains belonging to nine species of the Lactobacillus acidophilus rRNA-group have been analysed by SDS-PAGE of cellular proteins, RAPD-PCR and AFLP with fluorescently labeled primers in order to find improved methods for their identification. Strains of the following phenotypically highly similar species were examined: L. acidophilus, L. amylovorus, L. crispatus, L. johnsonii, L. gasseri, L. gallinarum, L. helveticus, L. iners and L. amylolyticus. Although the majority of the species can be differentiated by SDS-PAGE of whole-cell proteins, the latter technique showed poor discrimination between L. gasseri and L. johnsonii strains and between some strains of L. amylovorus and L. gallinarum. However, this study shows that the RAPD-PCR (using at least 3 different primers followed by numerical analysis of the combined patterns) and AFLP are most suitable genomic fingerprinting techniques for the differentiation of all the species listed above, and that databases for identification can be constructed, particularly when commercially available molecular tool-kits are used. The separate species status of the recently described L. amylolyticus and L. iners was fully confirmed. PMID- 10794144 TI - Thermodesulfobacterium hveragerdense sp. nov., and Thermodesulfovibrio islandicus sp. nov., two thermophilic sulfate reducing bacteria isolated from a Icelandic hot spring. AB - Two thermophilic non-sporeforming sulfate-reducing bacteria (SRB) were isolated from microbial mats collected from an Icelandic hot spring. Strain JSP was a gram negative rod, with an average cell size of 2.8 x 0.5 microm. No flagella were found. Growth occurred between 55 and 74 degrees C with an optimum between 70 and 74 degrees C at pH 7.0. The G+C content was 40 mol%. Strain R1Ha3 was a gram negative vibrio-shaped rod with an average cell size of 1.7 x 0.4 microm. Motility was observed mediated by one polar flagellum. The growth optimum at pH 7.0 was 65 degrees C, and growth occurred between 45 and 70 degrees C. The G+C content was 38 mol%. In the presence of sulfate, both strains used lactate, pyruvate and H2 as electron donors. In addition, strain R1Ha3 used formate. Pyruvate was the only substrate supporting fermentative growth of both strains. Growth occurred with sulfate as well as thiosulfate as electron acceptors. Furthermore, strain R1Ha3 reduced nitrate and strain JSP reduced sulfite. Neither of the strains were able to oxidize lactate completely to CO2 and neither of the strains contained desulfoviridin. 16S rDNA sequencing placed strain JSP in the genus Thermodesulfobacterium and strain R1Ha3 in the genus Thermodesulfovibrio. Based on the DNA-DNA hybridization studies and differences in morphology and physiology to their closest relatives the two new isolates were considered as new species. Strain JSP is named Thermodesulfobacterium hveragerdense and strain R1Ha3 Thermodesulfovibrio islandicus. PMID- 10794147 TI - A polyphasic taxonomic study of Chryseobacterium strains isolated from dairy sources. AB - A polyphasic taxonomic study, employing protein electrophoresis (SDS-PAGE), gas chromatographic analysis of cellular fatty acids (FAME), mol% G+C determination and DNA-DNA hybridizations, was undertaken on 103 dairy isolates shown to belong to Chryseobacterium. Reference strains of the Chryseobacterium species, CDC group IIb and Embedobacter brevis were included. SDS-PAGE analysis yielded good differentiation between the investigated species. About half of the strains could be clustered into nine major groups while the other half occupied a separate position. With FAME analysis no clear differentiation of the Chryseobacterium species (except C. meningosepticum) and SDS-PAGE groups could be achieved. FAME analysis, however, gave good differentiation between the Chryseobacterium and Empedobacter strains. The mol% G+C of the isolates tested, ranged between 36.4 and 39.0. The combination of SDS-PAGE and DNA-DNA hybridization identified a large group of dairy isolates as C. indologenes, one isolate as C. gleum and two new genotypic groups, comprising five and 15 dairy isolates respectively, emerged from the polyphasic study. Another large part of strains have a separate or uncertain position in Chryseobacterium and remained classified as Chryseobacterium species CDC group IIb. PMID- 10794145 TI - Methylomonas scandinavica sp. nov., a new methanotrophic psychrotrophic bacterium isolated from deep igneous rock ground water of Sweden. AB - Methane-utilizing bacteria were enriched from deep igneous rock environments and affiliated by amplification of functional and phylogenetic gene probes. Type I methanotrophs belonging to the genera Methylomonas and Methylobacter dominated in enrichment cultures from depths below 400 m. A pure culture of an obligate methanotroph (strain SR5) was isolated and characterized. Pink-pigmented motile rods of the new isolate contained intracytoplasmic membranes as stacks of vesicles, assimilated methane via the ribulose monophosphate pathway and had an incomplete tricarboxylic acid cycle. Phosphatidyl glycerol, methylene ubiquinone and cytochrome c552 were prevailing. The DNA G+C content is 53.3 mol %. Strain SR5 grew at temperatures between 5 and 30 degrees C with optimum at 15 degrees C, close to its in situ temperature. Analyses of 16S rRNA gene, whole cell protein, enzymatic and physiological analyses of strain SR-5 revealed significant differences compared to the other representatives of Type I methanotrophs. Based on pheno- and genotypic characteristics we propose to refer the strain SR5 as to a new species, Methylomonas scandinavica. PMID- 10794148 TI - Genetic relationships within the genus Prevotella analyzed by multilocus enzyme electrophoresis and DNA-DNA hybridization. AB - The genetic diversity and relationships within the genus Prevotella were studied by analyzing twenty-five strains by multilocus enzyme electrophoresis (MLEE) at nine metabolic enzyme loci and DNA-DNA hybridization. MLEE revealed a high genetic diversity with 25 electrophoretic types (ETs) for the 25 strains studied, a mean number of alleles per enzyme locus of 6.8 and a mean genetic diversity per locus of 0.786. The index of association described by Maynard Smith et al. (1993) revealed a clonal structure within the genus Prevotella. A dendrogram generated by cluster analysis of a matrix of ETs showed that species like P. bivia, P. buccae, P. oris, P. oralis, P. nigrescens, and P. denticola form clusters that are consistent with DNA homologies. However, strains identified as P. melaninogenica or P. loescheii by DNA-DNA hybridization did not constitute distinct subpopulations in MLEE. MLEE analysis demonstrated its high power in differentiating closely related strains. It provides an alternative to 16S rRNA analysis for the study of phylogenetic relationships within the genus Prevotella, especially for differentiating strains with high DNA homology or high rRNA homology. PMID- 10794149 TI - Numerical taxonomy of gram-negative, facultative anaerobic bacteria isolated from skin of turbot (Scophthalmus maximus) and surrounding water. AB - A numerical taxonomic study of 473 gram negative heterotrophic facultative anaerobic bacteria isolated from skin of turbot (Scophthalmus maximus) and its culture water was performed. The study included 53 type and reference strains belonging to the genera Vibrio, Aeromonas and Listonella. The strains were characterized using 90 tests and data were examined by Simple Matching coefficient (S(SM)) and Jaccard coefficient (S(J)). UPGMA (unweighted pair group method, arithmetic average) defined 66 phena at S(SM) values > or = 84% and 27 groups at S(SM) > or = 80%. Six phena were defined as Vibrio albensis, V. (Listonella) anguillarum, V. splendidus biotype I, V. fischeri, V. ordalii and V. scophthalmi including reference strains. Some groups clustered different phena for one species, although others as the V. anguillarum related strains and inactive Vibro group required S(SM) > or = 84% to define species. More studies are necessary to identify the Vibrio spp. strains and to confirm some species identifications. PMID- 10794150 TI - Kurtzmanomyces insolitus sp. nov., a new anamorphic heterobasidiomycetous yeast species. AB - A new anamorphic heterobasidiomycetous yeast species, Kurtzmanomyces insolitus, is described using a polyphasic taxonomic approach. The new species has the salient characteristics of the genus Kurtzmanomyces and, additionally, the ability to produce ballistoconidia. Data derived from comparative micromorphological studies, physiological characterisation, ultrastructure and nucleic acid analyses led to assigning the new species to Kurtzmanomyces rather than to the currently accepted genera of ballistoconidia-forming fungi. An emendation of the genus Kurtzmanomyces is proposed to allow the inclusion of the new species. PMID- 10794151 TI - Characterization of microbial communities of biofilters by phospholipid fatty acid analysis and rRNA targeted oligonucleotide probes. AB - The microbial community of a biofilter for waste gas treatment of an animal rendering plant was characterized by the analyses of the phospholipid fatty acids (PLFAs) of the filter material. For these analyses five samples of one filter were taken in intervals between one and two months. The main components of the PLFA profiles were straight chain saturated, monounsaturated and cyclopropyl fatty acids. Terminally branched and 10-methyl branched fatty acids were present in minor amounts. The structure and succession of the microbial community was interpreted by the presence and quantitative changes of diagnostic fatty acids. The stability of diagnostic fatty acids in relation to varying incubation parameters was tested for a number of bacterial isolates from biofilters representing different phylogenetic branches. For two samples, the data from the PLFA-analyses were compared with data obtained by hybridization with fluorescently labeled, rRNA-targeted oligonucleotide probes specific for the alpha-, beta- and gamma-subclass of the Proteobacteria, the Actinobacteria (Firmicutes with high G+C content) and the Firmicutes with low G+C content. These data indicated a dominating number of Proteobacteria (54% and 35% of DAPI-stained cells), in which the gamma-Proteobacteria represented the main fraction. Actinobacteria were detected in minor amounts, the number of Firmicutes with low G+C content was near the detection limit of the method. About half of the cells detected with a probe specific for Bacteria did not hybridize with the probes specific for the alpha-, beta- and gamma subclass of the Proteobacteria and the two subgroups of the Firmicutes. The results of both methods, the fluorescence in situ hybridization (FISH) and the PLFA analyses corresponded well and were best suited to confirm and complement each other. PMID- 10794152 TI - Polyphasic classification of 0.2 microm filterable bacteria from the western Mediterranean Sea AB - The 0.2 microm filtration of sea water samples from the Mediterranean Sea (Bay of Calvi, Corsica), collected from 10 m and 35 m depth led to the isolation of several gram-negative bacterial strains able to grow on full-strength media as well as on diluted media. The analysis of the 16S rRNA gene sequences and estimation of the phylogenetic relationships of these facultative oligotrophic bacteria indicated that they grouped into two phylogenetic branches. The strains RE10F/2, RE10F/5 (10 m depth samples) and RE35/F12 (35 m depth samples) were assigned to the gamma-subclass, while RE35F/1 (35m depth sample) was assigned to the alpha-4-subclass of the Proteobacteria. The strains RE10/F2 and RE10/F5 were most closely related to species and strains of the Pseudoalteromonas group, whereas the strain RE35F/12 placed adjacent to the family Vibrionaceae. The phylogenetic analysis of strain RE35F/1 revealed that this bacterium clusters with marine strains and species of the aerobic anoxygenic phototrophic bacteria Erythrobacter as well as Erythromicrobium and more distantly to Sphingomonas spp. Supplementary to those genotypic classifications the chemotaxonomic signatures including the major respiratory lipoquinone systems, the cellular fatty acid compositions as well as the polyamine contents of the bacteria were investigated. The isolated organisms displayed differences in their physiological and biochemical properties to already described strains belonging to the same genera or families, as revealed by the comparative 16S rRNA analysis. Despite the fact that these bacteria were isolated from a 0.2 microm filtrate, the cultured organisms which were all rod-shaped, displayed width dimensions ranging from 0.4 up to 0.7 microm, indicating that these bacteria were starvation forms at the time of isolation and not ultramicrobacteria as defined by Torella and Morita (1981) or by Schut et al. (1993). Because our isolated strains represent potentially new taxa, this first investigation on 0.2 pm filterable bacteria from the Western Mediterranean Sea supports the hypothesis that this bacterial fraction contributes to the diversity of marine bacteria. PMID- 10794153 TI - Diversity of bradyrhizobia from 27 tropical Leguminosae species native of Senegal AB - We isolated 71 slow-growing bacterial strains from nodules of 27 native leguminous plants species in Senegal (West-Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia playing an ecological role and having agronomic potential in arid regions. The isolates were characterised by restriction fragment length polymorphism (RFLP) analysis of PCR amplified 16S rDNA and comparative SDS-PAGE of whole-cell proteins; reference strains of the different known rhizobial species and groups were included as references. We conclude that these nodule isolates are diverse, and form several phylogenetic subgroups inside Bradyrhizobium. Nodulation tests performed on 5 plant species demonstrated host specificity among the strains studied. PMID- 10794155 TI - Clarity in the clinical medical literature: we are no longer preaching to the choir. PMID- 10794154 TI - In vitro susceptibilities of Aeromonas genomic species to 69 antimicrobial agents. AB - A total of 217 strains representing all 14 currently described genomic species in the genus Aeromonas were tested for susceptibility to 69 antimicrobial agents by a microdilution method. All species were susceptible to tetracyclines, quinolones, chloramphenicol, and most of the aminoglycosides and the cephalosporins, but were resistant to lincosamides, vancomycin, teicoplanin and some penicillins. In general, no significant differences were found that correlated with the taxonomic designation or the origin of the isolates tested. The microdilution method proved to be easy to perform allowing susceptibility testing of extensive strain collections for a large number of antimicrobial agents. PMID- 10794156 TI - Ancillary methods for the detection of recurrent urothelial neoplasia. PMID- 10794157 TI - Subdividing atypical glandular cells of undetermined significance according to the Australian modified Bethesda, system: analysis of outcomes. AB - BACKGROUND: The Australian modification of the Bethesda system (TBS) for reporting endocervical cellular abnormalities differs from TBS in having a "high grade" category for prediction of adenocarcinoma in situ (AIS), an "inconclusive" category requiring that AIS be excluded (AISEX), and a "low grade" category (Enatyp) for when only minor changes are seen in endocervical cells. METHODS: From subsequent follow-up, the authors evaluated the proportion of histologically confirmed high grade lesions (squamous, glandular, or endometrial) in each cytologic reporting category, for cases in which no concurrent squamous abnormality was also predicted. RESULTS: From information was available for 95.9% of 172 AIS predictions, 81.5% of 119 AISEX predictions, and 89.0% of 237 reports of Enatyp. The histologic positive predictive values for high grade lesions were 95.7% for AIS, 71.1% for AISEX, and 7.1% for Enatyp. In the first 2 categories, a minority of lesions proved to be purely squamous (3.8% for AIS and 33.3% for AISEX), whereas in the Enatyp category 87.0% were squamous only. CONCLUSIONS: Compared with TBS, the Australian terminology for reporting of endocervical abnormalities better reflects the risk of underlying high grade abnormalities and may result in more appropriate management recommendations. For Enatyp reports, which constitute over 85% of reported endocervical abnormalities, repeat cytology should be considered appropriate management. However, cone biopsy should be the investigative method of choice for women with reports of AIS and AISEX. PMID- 10794158 TI - Invasive carcinoma in clinically suspicious breast masses diagnosed as adenocarcinoma by fine-needle aspiration. AB - BACKGROUND: Fine-needle aspiration (FNA) biopsy of palpable breast masses along with clinical and radiologic findings can provide rapid distinction between benign and malignant lesions. A preoperative determination of invasive or in situ carcinoma assists in the planning of definitive treatment. Previous studies have concentrated on whether cytologic features adequately distinguish invasion, but to the authors' knowledge the predictive value of clinicopathologic correlation has not been investigated. The authors attempted to determine whether a malignant cytologic diagnosis for a palpable breast mass is sufficient for its definitive surgical management as an invasive neoplasm. METHODS: The authors reviewed 351 FNAs from palpable breast lesions with a cytologic diagnosis of "adenocarcinoma." The presence of invasive disease was determined by histologic demonstration of invasive carcinoma in the corresponding surgical specimen or by identifying metastatic carcinoma in the absence of another primary source. RESULTS: Three hundred forty-three (97.7%) palpable tumors diagnosed as adenocarcinoma by FNA proved to be invasive adenocarcinoma. The remaining eight tumors contained high grade ductal carcinoma in situ, and two of these contained foci suggestive of microinvasion. CONCLUSIONS: A palpable breast mass with an FNA diagnosis of adenocarcinoma usually represents invasive carcinoma. A definitive treatment plan therefore can be planned based on these clinical and FNA findings. PMID- 10794159 TI - Adhesion of aspirated tumor cells to extracellular matrix proteins: a new methodology utilizing fine-needle aspiration. AB - BACKGROUND: Cell adhesion molecules mediate the interactions of cells with other cells and with extracellular matrix components. Such interactions may be important in the development of tumor invasion and metastasis. This article describes a new approach to the evaluation of tumor cell-matrix interactions by utilizing fine-needle aspiration of resected tumors. METHODS: Fine-needle aspiration was performed on 15 fresh surgical specimens of various types of carcinomas. After partial purification by isotonic Percoll centrifugation, tumor cell adhesion to collagen Type IV, laminin, and fibronectin was evaluated by counting cytologically malignant cells adhering to matrix-coated plastic substrates. Frozen tissue sections of the corresponding tumors were studied simultaneously for immunohistochemical expression of alpha-2, alpha-3, alpha-4, and alpha-5 integrin subunit expression. Results of the immunohistochemical staining then were compared with the adhesion data for particular tumors. RESULTS: In general, the majority of the tumors exhibited little or no adhesion to collagen or laminin, but several tumors showed marked adhesion to fibronectin. Striking differences were noted between some tumors of the same histologic subtype. Competitive inhibition studies performed with two of the tumors (a large cell carcinoma and a renal cell carcinoma) showed decreased adhesion to fibronectin in the presence of anti-alpha-5, suggesting at least a partial role for the alpha-5-beta 1 fibronectin receptor in mediating the adhesion of these tumors to fibronectin. All the tumors examined exhibited strong immunohistochemical expression of the alpha-2 and alpha-3 integrin subunits, and all were negative for alpha-4. Three of the tumors showed weak expression of alpha-5, two of which (a squamous cell carcinoma and a renal cell carcinoma) were the tumors that showed the greatest adhesion to fibronectin. CONCLUSIONS: Quantitative adhesion data can be obtained using cell suspensions prepared from fine-needle aspirates, and there are marked differences in adhesive properties between particular tumors. Although two of the tumors showed a correlation between adhesion to fibronectin and immunohistochemical expression of the alpha-5 integrin subunit, matrix adhesion does not necessarily correlate with immunohistochemical expression of adhesion molecule receptors. In the future, this methodology potentially could be of value in determining which patients may benefit from therapies aimed at modifying tumor cell-matrix interactions. PMID- 10794160 TI - Fine-needle aspiration of cystic parotid glands lesions: an institutional review of 46 cases with histologic correlation. AB - BACKGROUND: Many case reports describing various benign and malignant disease entities diagnosed in cystic parotid gland lesions by fine-needle aspiration (FNA) exist in the literature. Very few studies, however, discuss the efficacy and the diagnostic difficulties of FNA cytology in such lesions. The authors report a 5-year institutional experience with FNA cytology of cystic parotid gland lesions and address the accuracy of the procedure and avoidance of diagnostic pitfalls. METHODS: A retrospective review of 46 cases in which patients clinically presented with cystic parotid gland lesions was done from a total of 221 parotid FNAs performed over a 5-year period at the University of Pennsylvania Medical Center. The clinical features, cytology, and follow-up surgical pathology were reviewed to determine diagnostic accuracy and highlight potential pitfalls. RESULTS: Based on subsequent surgical excision in 29 of the 46 cases of cystic parotid gland lesions, a diagnostic accuracy rate of 83% was obtained for FNA. Approximately 20% (9 of 46 cases) were found to be clinically significant tumors and included 4 mucoepidermoid and 1 adenocarcinoma (NOS), 3 benign mixed tumors, and 1 metastatic melanoma. Three major factors were implicated as diagnostic pitfalls in our series: 1) failure to obtain critical clinical information, 2) overinterpretation of paucicellular specimens, and 3) failure to realize that certain tumors generally perceived as solid masses can present clinically as cystic lesions. CONCLUSIONS: FNA cytology is a valuable tool in the primary diagnosis and management of cystic parotid gland lesions. The diagnostic accuracy of this procedure can be significantly improved by acquiring a detailed clinical history, obtaining an adequate cellular specimen, and having knowledge of the variety and frequencies of possible diagnostic entities that may present as cystic parotid gland lesions. PMID- 10794161 TI - Correlation between morphology and telomerase activity in cells from exfoliative lung cytologic specimens. AB - BACKGROUND: Telomerase is a ribonucleoprotein that compensates for the erosion of telomeres (chromosomal termini). Telomerase activity is detected in more than 85% of cancerous lesions and is therefore considered a novel marker of cancer. The authors compared cytologic morphology and telomerase activity at the cellular level to obtain further insight into their association. METHODS: The authors used bronchial washing and brushing materials obtained from 18 patients with lung carcinomas (6 squamous cell, 8 adenocarcinoma, 2 large cell, 1 small cell, and 1 metastasis from colon carcinoma) and 20 patients with nonmalignant disease. An in situ telomeric repeat amplification protocol (TRAP) assay was performed, and routine Papanicolaou-stained slides using the same sample were assessed. RESULTS: Nuclear fluorescent signals at the nuclear area, corresponding to telomerase activity, shown by the in situ TRAP assay were only detected in samples containing morphologically malignant cells. No nuclear fluorescence was seen in the keratinizing component of well-differentiated squamous cell carcinoma. Nuclear staining was not seen in metaplastic or basal hyperplastic cells. Cytoplasmic fluorescence was only found in macrophages and polymorphonuclear leukocytes. CONCLUSIONS: Nuclear fluorescence corresponding to telomerase activity was not demonstrated in metaplastic or basal hyperplastic cells, thus indicating that detection of telomerase activity is closely associated with the presence of malignant cells, but not premalignant lesions, in lung carcinoma patients. Moreover, in some samples with cancer, cells failed to show telomerase activity, suggesting the limitation of this method for the detection of malignant cells in certain lung carcinoma patients. PMID- 10794162 TI - Numeric chromosomal abnormalities in small lymphocytic and transformed large cell lymphomas detected by fluorescence in situ hybridization of fine-needle aspiration biopsies. AB - BACKGROUND: Chromosomal abnormalities in some lymphomas are associated with a poor prognosis. Trisomy 12 and deletions of chromosome 13 have been described in small lymphocytic lymphoma (SLL). To determine whether chromosomal aberrations could be detected by fluorescence in situ hybridization (FISH) on fine-needle aspiration biopsies (FNABs), we analyzed and compared specimens from seven patients with SLL and nine patients with a history of SLL that had transformed to large cell lymphoma. METHODS: DNA probes specific to chromosome 12 and the chromosome 13/RB-1 gene locus were used for in situ hybridization on interphase cell nuclei. The cytologic features, ploidy, proliferation index, and conventional cytogenetic analysis findings were correlated with the FISH results. RESULTS: Trisomy 12 was detected in 2 (29%) of the 7 cases of SLL and 2 (22%) of the 9 cases of transformed large cell lymphoma. Deletion of chromosome 13/RB-1 was present in 3 (43%) of 7 cases of SLL and 3 (33%) of 9 cases of transformed large cell lymphoma. CONCLUSIONS: This study showed that interphase cytogenetic analysis by FISH is feasible on FNAB specimens. Trisomy 12 and deletions of chromosome 13/RB-1 were present in some cases of SLL and transformed large cell lymphoma, and the presence of these chromosomal abnormalities did not correlate with transformation to a higher grade of lymphoma. PMID- 10794163 TI - Alteration of cardiac myofibrillogenesis by liposome-mediated delivery of exogenous proteins and nucleic acids into whole embryonic hearts. AB - A precise organization of contractile proteins is essential for contraction of heart muscle. Without a necessary stoichiometry of proteins, beating is not possible. Disruption of this organization can be seen in diseases such as familial hypertrophic cardiomyopathy and also in acquired diseases. In addition, isoform diversity may affect contractile properties in such functional adaptations as cardiac hypertrophy. The Mexican axolotl provides an uncommon model in which to examine specific proteins involved with myofibril formation in the heart. Cardiac mutant embryos lack organized myofibrils and have altered expression of contractile proteins. In order to replicate the disruption of myofibril formation seen in mutant hearts, we have developed procedures for the introduction of contractile protein antibodies into normal hearts. Oligonucleotides specific to axolotl tropomyosin isoforms (ATmC-1 and ATmC-3), were also successfully introduced into the normal hearts. The antisense ATmC-3 oligonucleotide disrupted myofibril formation and beating, while the sense strands did not. A fluorescein-tagged sense oligonucleotide clearly showed that the oligonucleotide is introduced within the cells of the intact hearts. In contrast, ATmC-1 anti-sense oligonucleotide did not cause a disruption of the myofibrillar organization. Specifically, tropomyosin expression can be disrupted in normal hearts with a lack of organized myofibrils. In a broader approach, these procedures for whole hearts are important for studying myofibril formation in normal hearts at the DNA, RNA, and/or protein levels and can complement the studies of the cardiac mutant phenotype. All of these tools taken together present a powerful approach to the elucidation of myofibrillogenesis and show that embryonic heart cells can incorporate a wide variety of molecules with cationic liposomes. PMID- 10794164 TI - Proliferation and differentiation of pituitary corticotrophs during the fetal and postnatal period: a quantitative immunocytochemical study. AB - To study the proliferation and differentiation of pituitary corticotrophs, we administered bromodeoxyuridine (BrdU) to pregnant rats at 15.5-21.5 days of gestation and to rat pups at 3, 7, and 28 days after birth. The pituitary sections of fetuses and pups were consecutively immunostained with anti-BrdU and anti-adrenocorticotropic hormone (ACTH) to detect proliferating cells and corticotrophs, respectively. The number of cells labeled with BrdU, ACTH, or both were counted. The diameters of their nuclei and the volume of the pituitary were measured. The BrdU-positive cells were around 76,000-96,000/mm3 during the period studied. The corticotrophs were first detected in the fetus at 15.5 days and they increased during the fetal and postnatal periods. The double-labeled cells were first detected in the 17.5-day fetus. They increased markedly at 19.5 days and comprised about one-quarter of the corticotrophs that increased in 24 h at this stage. These results indicate that: (1) at 15.5-18.5 days the corticotrophs were derived almost exclusively from undifferentiated cells; (2) during the later fetal and early postnatal periods, the proliferation of existing corticotrophs contributed, at least in part, to their increase; (3) about 1/20 of proliferating cells differentiated to corticotrophs when their increase was required. PMID- 10794165 TI - Reconstructing the pathway of the tensor veli palatini motor nerve during early mouse development. AB - The motor axons innervating the tensor veli palatini (TVP) navigate a long distance from the trigeminal motor nucleus to their target. The pathway and time course of the TVP motor nerve during this navigation process remain poorly understood. The aim of this study was to elucidate the peripheral development of the TVP motor nerve, and to confirm when the morphological relationship is established between the nerve and target muscle progenitors. Using immunohistochemistry, carbocyanine fluorescent labeling, and computerized three dimensional image-reconstruction methods, we demonstrated the development of the TVP motor nerve in mouse embryos. Further, the morphological relationship between the extending mandibular nerve and myogenic cells stained for MyoD1 was examined. The peripheral pathfinding of the TVP motor nerve was divided into three continuous stages: (1) the earliest trigeminal motor axons leave the metencephalon and enter the primordium of the trigeminal ganglion at E9.5, when MyoD1-positive cells can already be detected in the mesenchymal core of the mandibular arch; (2) converging with the sensory root, the trigeminal motor root excites the trigeminal ganglion and begins to approach the mandibular muscle precursors at E10.5; (3) collateral branching occurs at E12.5. By E13.5, a nerve branch splits from the mandibular nerve to innervate the TVP, which appears as an individual muscle mass. These results suggest that the early process of mandibular motor nerve extension is correlated with the trigeminal ganglion cells, whereas when growing out of the ganglion, the mandibular nerve has a close relationship with target myogenic cells throughout the later process of pathway finding. PMID- 10794166 TI - Study of the olivocochlear neurons using two different tracers, fast blue and cholera toxin, in hypothyroid rats. AB - Congenital hypothyroidism results in deafness that is caused by changes in the auditory receptor, including scanty development of the outer hair cells and a lack of synaptogenesis between these cells and the efferent system. although the afferent population is present. The normal efferent innervation of the cochlea originates in the superior olivary complex, arising from efferent neurons belonging to the lateral or to the medial olivocochlear system. In the rat, the former is constituted by neurons located in the lateral superior olivary nucleus, that project to the inner hair cells, while the later originates in the ventral nuclei of the trapezoid body and project to the outer hair cells. The aim of this work is to study the localization, number and morphology of the olivochochlear neurons in congenital hypothyroid animals by means of the injections of the retrograde tracers, either fast blue or cholera toxin, in the cochlea. The mean total number of labeled olivocochlear neurons after injection of fast blue in hypothyroid animals was 1,016, and in control ones was 1,027. Using cholera toxin, the mean total number of labeled olivocochlear neurons was slightly lower: 863 in hypothyroid animals versus 910 in control ones. Although both tracers showed no significant differences between groups, when the somatic area of the labeled olivocochlear neurons is considered, the size of all of the three different population of cells (lateral olivocochlear neurons, medial olivocochlear neurons and shell neurons) was significantly lower in the hypothyroid rats. This is the first study of the olivocochlear neurons in hypothyroid animals. The conclusion from this work is that in hypothyroid rats the labeled olivocochlear neurons are significantly smaller but that there is not any modification in the localization and number of the labeled olivocochlear neurons, suggesting that thyroid hormones are necessary for the neuronal growth. However, most of the medial olivocochlear neurons do not make contact with their target, so their maintenance suggests that the axons are in contact with other structures of the cochlea. PMID- 10794167 TI - Glycine, glycine receptor subunit and glycine transporters in the rat parabrachial and Kolliker-Fuse nuclei. AB - In the present study, we investigated the expression and distribution of key molecules in the parabrachial (PB) and Kolliker-Fuse nuclei (KF) that determine glycinergic signal transduction. By means of immunocytochemistry, we analyzed the amino acid glycine (Gly), the glycine transporters 1 and 2 (GlyT1, GlyT2), and the ligand binding glycine receptor-subunit alpha 1 (GlyR alpha 1). Gly immunoreactivity (-ir) was mainly found in varicose fibers and presumed terminal boutons; Gly-ir cell bodies were only occasionally seen. Immunoreactivity for GlyT2 was located in axons while GlyT1-staining was diffuse in the neuropil. Immunolabeling for GlyR alpha 1 occurred mostly as granular staining diffusely distributed throughout the neuropil. Only in the superior lateral PB, the lateral crescent of the PB, and caudally in the KF did GlyR alpha 1-ir outline cell bodies and primary and higher-order dendrites. Furthermore, our data demonstrate a distinct codistribution of immunoreactivities for Gly, GlyT2. and GlyR alpha 1 in a specific set of PB nuclei and in the KF. Strong staining was consistently seen in the internal lateral PB, the ventral lateral PB, the lateral crescent, the medial PB adjacent to the superior cerebellar peduncle, and the rostral two thirds of the KF. Moderate to weak immunostaining was present in the superior, central, and dorsal lateral PB, the external medial PB, the medioventral part of the medial PB, and caudally in the KF. In contrast, remaining nuclei such as the external lateral PB and the waist area were essentially devoid of Gly-ir profiles, GlyT2-ir, and GlyR alpha 1-ir. Immunoreactivity for GlyT1 was evenly distributed throughout all nuclei of the medial and lateral PB, including the external lateral PB and the waist area, while the KF was only weakly stained. Our data provide evidence that glycinergic mechanisms might play a role for neural processing in most nuclei of the PB and in the KF. Only the external lateral PB and the waist area are apparently not subject to glycinergic inhibition. PMID- 10794168 TI - The significance of rudimentary nephrostomial tubules for the origin of the vertebrate gonad. AB - Initial gonadal development was studied in 30- to 40-day-old bovine embryos. The results were interpreted in conjunction with findings on pro- and mesonephric organization in larval forms of Ichthyophis kohtaoensis (Gymnophiona, Amphibia). In bovine embryos vestigial nephrostomial tubules are the immediate precursors of the blastemas for adrenocortical, rete, gonadal and Mullerian infundibular development. From the study of Ichthyophis it can be concluded that the vestigial nephrostomial tubules seen in the bovine embryo are pronephric and not mesonephric in nature. As a consequence, the indifferent mammalian gonad is defined as a modified homologue of the pronephros situated in the zone of pro /mesonephric overlapping. Such an overlapping of the two kidney generations in the fully developed state is clearly seen in Ichthyophis. Overlapping of the mesonephros with the modified pronephros (gonad) is necessary to allow intercalation of mesonephric tubules (efferent ductules in mammals) into the male seminal excurrent duct system. PMID- 10794169 TI - Pre-eclampsia associated alterations of the elastic fibre system in umbilical cord vessels. AB - Although pre-eclampsia (PE) is often associated with fetal hypoxia, hypertension and/or disturbed function of the fetal circulation, the effect of these altered hemodynamic parameters on the structure and composition of umbilical vessels has not been systematically investigated before. Therefore, this study focuses on PE associated changes of the elastic fibre system in umbilical cord vessels investigated by light and electron microscopy, immunocytochemistry and biochemistry. In umbilical cord veins, no changes in thickness of the vessel wall or of any sublayer were observed. However, the internal elastic lamina of the veins was split in 80% of the PE-group in contrast to 20% in uncomplicated pregnancies. This effect was significant (alpha <0.01) from 36 weeks of gestation onwards. In umbilical cord arteries, the entire arterial vessel wall was found to be 15% thicker in PE than in uncomplicated pregnancies. The enlargement was caused by an increase of both the tunica intima and tunica media. The thickening of the tunica intima was attributed to a migration of smooth muscle cells towards the endothelium, accompanied by a splitting of the internal elastic lamina. Compared to uncomplicated pregnancies, smooth muscle cells of arteries and veins in PE showed a metabolic activation demonstrated by highly dilated endoplasmic reticulum. A semiquantitative score method as well as a quantitative dot blot assay indicated a PE-associated reduction of elastin expression in the arterial vessel walls. In summary, PE obviously induces a decrease of the elastin content accompanied by a thickening of the vessel wall in umbilical cord arteries. This remodeling of the elastic fibre system, together with an increased migration of smooth muscle cells, might represent part of the functional adaptation system of the umbilical cord arteries on the altered hemodynamic conditions in PE. PMID- 10794170 TI - Structure of the cone photoreceptor mosaic in the retinal periphery of adult humans: analysis as a function of age, sex, and hemifield. AB - We have analyzed variation in the structure of the human photoreceptor mosaic as a function of age, sex, and retinal location using the nearest neighbor and density recovery profile methods. In contrast to most previous work, we have focused our analysis on the mid- and far-peripheral human retina. Video-enhanced differential interference contrast optics was used to characterize differences in the nasal and temporal periphery of unstained wholemounts from 12 males and 12 females ranging in age from 15- to 83-years-old. At sites matched for cone density (approximately 5,000 cones/mm2), the mosaic is far more orderly in the temporal than in the nasal periphery. This is true at all ages and in both sexes. Despite their increased local order, regularity ratios of adjacent temporal fields tend to be much more variable than are those of adjacent nasal fields. These marked nasal-temporal differences are eliminated when eccentricity is held constant and cone density is allowed to vary. There is a mild, statistically significant age-related decline in the regularity of the cone mosaic, but only in the nasal periphery. There are no significant differences in the precision of the cone mosaic between sexes. The equivalence of the regularity of the mosaic at matched eccentricities, but not at matched cone densities, suggests that the irregularity of the mosaic is secondary to developmental gradients and, more generally, to reduced selection for high acuity vision in the retinal periphery. PMID- 10794171 TI - Distribution of fibrillin I in extracellular matrix and epithelia during early development of avian embryos. AB - Fibrillin microfibrils are widely distributed components of extracellular matrices that function in the formation of elastin, serve structural roles and provide substrates for cell adhesion. To determine when and how fibrillin-1 (fib 1) may function in early development we have examined the temporal and spatial distribution of fib-1 in chicken embryos. Using homologous PCR we amplified and cloned a 407 nt fragment of chicken cDNA that appears to code for an orthologue of FBN-1. Bacterially expressed protein was used to prepare two monoclonal antibodies, both of which recognize a 350 kD band in immunoblots or immunoprecipitates in supernatants of chicken embryonic aorta cells or human MG 63 cells. Both antibodies recognize fibrillar material associated with the surfaces of cultured cells. The antibodies appear to be specific for fib- as there was only weak cross reactivity to a bacterially expressed fragment from the corresponding region of fib-2 and the pattern of immunofluorescence in embryonic tissue is distinctly different from that of JB-3, a fib-2 specific antibody (Rongish et al. 1998). In embryos, fib-1 is first detected at stage 6 in the epiblast during gastrulation. In subsequent stages fib-1 fibers appear in all tissues and are present throughout the first 6 days of development. Immunoreactive fibers are present in basal laminae and mesenchyme filled spaces, but they also form random arrays with an apical-basal polarity within epithelia. Using primers specific for FBN-1 and FBN-2 in RT-PCR reactions we confirm the presence of fib- 1 and fib-2 mRNA in early embryonic stages. This temporal and spatial distribution indicates fib-1 has functions in early development that are distinct from fib-2. PMID- 10794172 TI - The Schizosaccharomyces pombe rfc3+ gene encodes a homologue of the human hRFC36 and Saccharomyces cerevisiae Rfc3 subunits of replication factor C. AB - In the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe replication factor C (RF-C) plays key roles both in chromosomal DNA replication and in DNA replication checkpoint function. At the replication fork, the five-subunit RF-C complex functions to load the trimeric polymerase accessory factor PCNA onto DNA. PCNA then acts as a sliding clamp, tethering Pol delta to the DNA to maximise its processivity. Here we describe the cloning of the S. pombe rfc3+ gene, encoding a homologue of the S. cerevisiae Rfc3 and human hRFC36 proteins. The 1026 bp rfc3+ ORF is interrupted by five introns, ranging in size from 49 to 165 bp. The spliced ORF is predicted to encode a 342 amino-acid protein that is approximately 50% identical at the amino acid sequence level to the S. cerevisiae Rfc3 and human hRFC36 proteins. As expected, S. pombe rfc3+ is an essential gene, with rfc3delta cells being defective for DNA replication. Loss of rfc3+ function can be rescued by heterologous expression of either the S. cerevisiae Rfc3 or human hRFC36 proteins in S. pombe. PMID- 10794173 TI - UBC13, a DNA-damage-inducible gene, is a member of the error-free postreplication repair pathway in Saccharomyces cerevisiae. AB - The Ubc13 protein was recently identified for its unique role in ubiquitin (Ub) chain assembly at the Ub Lys-63 residue instead of the conventional Lys-48 residue. This activity requires Ubc13 to form a complex with Mms2 and indeed ubc13 and mms2 mutations have been shown elsewhere to be epistatic with respect to UV sensitivity. The MMS2 gene is known to be a member of the error-free DNA postreplication repair (PRR) pathway. By contrast, the Ub Lys-63 residue has been previously implicated in the error-prone PRR pathway, since yeast cells carrying the ubiK63R mutation are defective in UV-induced mutagenesis. In the present study, we attempted to define the role of UBC13 within the PRR pathway. We found that the ubc13 mutation is epistatic to mms2 and rad6, confirming that UBC13 belongs to the PRR-pathway. We also found that ubc13 is synergistic to the error prone PRR pathway mutation rev3, indicating that UBC13 is in a pathway alternative to REV3 mutagenesis. The ubc13 mutant displays up to a 30-fold increase in the spontaneous mutation rate, and this increase is largely REV3 dependent. In addition, UV-induced mutagenesis is fully functional in the ubc13 mutant. These results together demonstrate that UBC13 is a member of the error free PRR pathway. The involvement of UBC13 in cellular tolerance to DNA-damage is further implicated by our finding that the UBC13 transcript level is increased up to 6-fold in response to DNA-damage. PMID- 10794174 TI - Glutathione, but not transcription factor Yap1, is required for carbon source dependent resistance to oxidative stress in Saccharomyces cerevisiae. AB - Resistance of haploid yeast to hydrogen peroxide and to tert-butylhydroperoxide strongly increases when 4% glucose is replaced by glycerol or ethanol as the carbon source of the complex medium. Using a GSH1-promoter-lacZ-fusion reporter construct we could demonstrate that GSH1 is one of the genes that are up regulated during the shift from fermentative to oxidative metabolism. A gsh1 mutant did not exhibit respiratory growth resistance to H2O2, whereas it was only slightly impaired in acquiring resistance against t-BOOH in the same experimental conditions. An isogenic deltayap1 mutant, although more sensitive to oxidative stress than the wild-type (WT), could increase resistance to both peroxides by a similar factor as observed for the WT when shifted from 4% glucose to a non fermentable carbon source. This indicates that in this case induction of resistance to oxidative stress is independent from Yap1 and from the Yap1 mediated stress response via the STRE motif. PMID- 10794175 TI - Evolutionary significance of an unusual chloroplast DNA inversion found in two basal angiosperm lineages. AB - Two basal lineages of flowering plants possess an intergenic inversion in the chloroplast inverted repeat (IR), a region of the genome from which there have been few previous reports of this class of structural mutation. The size of the inversion (approximately 200 bp) places it in a class not previously seen in the plastid genome. The two lineages with the rearrangement, representatives of the orders Laurales and Nymphaeales, are not closely related and the inversion therefore probably arose independently in each group. The inversion is bordered by short, but highly conserved, inverted repeat motifs that were most likely associated with the inversion process. A stem-loop structure that involves these motifs may play a functional role in mRNA stability. It is seen in all optimal or nearly optimal predicted RNA foldings of the intergenic region. PMID- 10794176 TI - The fungal acl1 and acl2 genes encode two polypeptides with homology to the N- and C-terminal parts of the animal ATP citrate lyase polypeptide. AB - ATP citrate lyase (ACL) catalyzes the formation of cytosolic acetyl-CoA, which is mainly used for the biosynthesis of fatty acids and sterols. In this paper, we show for the first time that in filamentous fungi two different subunits of ACL are encoded by two separate genes. This is in contrast to animals where ACL is encoded by a single gene. Data are presented on acl genes from the filamentous fungi Sordaria macrospora and Gibberella pulicaris. In S. macrospora, both genes, acl1 and acl2, are clustered within a region of 10 kb and are divergently transcribed. PMID- 10794177 TI - Methylation of the foreign transposon Restless in vegetative mycelia of Neurospora crassa. AB - Methylation of foreign and/or repeated sequences in the filamentous fungus Neurospora crassa is believed to be directed against invading transposable elements. To test this hypothesis, the fate of a transposon in N. crassa was investigated. Vectors were constructed which carried the transposon Restless, an active class-II element isolated from the fungus Tolypocladium inflatum. These vectors were introduced into N. crassa strains by protoplast transformation. Two strategies were employed: (1) ectopic multi-copy integration, and (2) site specific single-copy integration at the his-3 locus. All ectopic transformants exhibited strong methylation as confirmed by Southern hybridization of genomic DNA digested with the methylation-sensitive endonuclease Sau3AI and the methylation-insensitive endonuclease NdeII. Single copies of Restless integrated at the his-3 locus were not methylated. These results are discussed with respect to non-RIP methylation and potential consequences for gene-tagging strategies based on the use of Restless. PMID- 10794178 TI - Identification and characterization of PaMTH1, a putative O-methyltransferase accumulating during senescence of Podospora anserina cultures. AB - A differential protein display screen resulted in the identification of a 27-kDa protein which strongly accumulates during the senescence of Podospora anserina cultures grown under standard conditions. After partial determination of the amino-acid sequence by mass-spectrometry analysis of trypsin-generated fragments, pairs of degenerated primers were deduced and used to amplify parts of the sequence coding for the protein. These PCR products were utilized to select specific cDNA and genomic clones from DNA libraries of P. anserina. A subsequent DNA-sequence analysis revealed that the 27-kDa protein is encoded by a discontinuous gene, PaMth1, capable of coding for 240 amino acids. The first three amino-terminal residues appear to be removed post-translationally. The deduced amino-acid sequence shows significant homology to S-adenosylmethionine (SAM)-dependent methyltransferases. We hypothesize that the 27-kDa protein, PaMTH1, is involved in age-related methylation reactions protecting aging cultures against increasing oxidative stress. PMID- 10794179 TI - Carboxin resistance transformation of the homobasidiomycete fungus Pleurotus ostreatus. AB - A novel selection marker gene for transformation of the white-rot basidiomycete Pleurotus ostreatus was developed by introducing a point mutation in a gene which encodes the iron-sulfur protein (Ip) subunit of succinate dehydrogenase. The mutant gene, CbxR, encodes a modified Ip subunit with an amino-acid substitution (His239 to Leu) and confers resistance to the systemic fungicide, carboxin. The DNA sequence was integrated ectopically in the chromosome of the transformants. This is the first report of a homologous marker gene which is available for the molecular breeding of an edible mushroom. PMID- 10794180 TI - Determination of urinary and serum beta-glucuronidase and alkaline phosphatase in various renal disease and kidney rejection transplanted patients. AB - Beta-glucuronidase (beta-Glu) and alkaline phosphatase (ALP) were evaluated in serum and urine in 50 subjects classified into five equal groups. Group I was control healthy subjects, while groups II, III, IV, and V were patients with nephritic syndrome, pyelonephritis, kidney rejection, and end-stage renal disease, respectively. Urinary beta-Glu was significantly elevated in all four groups; while serum enzyme showed no change. On the other hand, serum ALP showed a significant elevation in all abnormal groups. Accordingly, urinary beta-Glu and serum ALP could be used as diagnostic markers for various renal diseases. PMID- 10794181 TI - Recombinant extracellular domain of rabbit growth hormone receptor and biological activity of somatogenic hormones. AB - The cDNA of the extracellular domain of rabbit growth hormone receptor (rbGHR ECD) was cloned in the prokaryotic expression vector pMON, to enable its expression in Escherichia coli after induction with nalidixic acid. The bacterially expressed rbPRLR-ECD protein, contained within the refractile-body pellet, was solubilized in 4.5 M urea, refolded, and purified on a Q-Sepharose column, pH 8, by stepwise elution with NaCl. The bioactive monomeric 28-kDa fraction was eluted in 0.15 M NaCl, yielding 50 mg/2.5 l of induced culture. The purified protein was over 98% homogeneous, as shown by SDS-PAGE in the presence or absence of reducing agent, and by chromatography on a Superdex column. Gel filtration was used to determine the stoichiometry of rbGHR-ECD's interaction with human (h), ovine (o), chicken (ch) and common carp (cc) GHs and with bovine (b) and caprine (c) placental lactogens (PLs). The formation of 2:1 complexes was indicated in all cases. Binding experiments using radiolabelled oGH as a ligand revealed it to be the most effective competitor, followed by bPL, cPL, hGH chGH and ccGH, with respective IC50 values of 0.27, 0.94, 1.55, 2.13, 41.9 and 51.2 nM. Rabbit GHR-ECD inhibited the bPL-inducible proliferation of FDC-P1 cells stably transfected with rbGHR and Nb2 cells possessing rat PRLR. The biological activity of oGH, hGH, cPL, bPL, chGH and ccGH was tested in the FDC-P1 cells stably transfected with rbGHR and yielded the respective EC50 values (in nM) of 0.024, 0.023, 0.021, 0.24, 4.71 and 0.49. These results indicate remarkable discrepancies between the binding capacities and biological activities: the possible reasons for these findings are discussed. PMID- 10794182 TI - A simplified procedure for purification of cytochrome P-450 by preparative ampholine gel for isoelectric focusing. AB - The purification process for cytochrome P450 is very complicated, involving five or more column chromatography steps for the final preparation. This paper describes a reduction in the number of the steps; it can be easily purified from pig testis microsomes with improved the yield. As the first step, DEAE-Toyopearl column chromatography is performed only once and then, as the second step, the partially purified cytochrome P450 is completely purified by a preparative Ampholine PAG-plate Gel for Isoelectric Focusing. The combination reduced the purification to a two-step procedure. PMID- 10794183 TI - Partial preparative purification of leptine I from foliage of the wild potato, Solanum chacoense (Bitt.). AB - Leptine I, a glycoalkaloid only known to occur in the foliage of the wild potato species Solanum chacoense (Bitt.), is a potent feeding deterrent to the economically serious insect pest, the Colorado potato beetle (Leptinotarsa decemlineata Say). In order to demonstrate, systematically, the effectiveness of leptine I, incorporation into synthetic beetle diet trials is necessary. We describe a preparative procedure for the partial purification of leptine I by a series of steps, starting with a solid-phase C18 extraction, followed by sequential silica gel chromatography, and finally reversed-phase preparative HPLC. This preparation yields a white powder, containing leptine I as the sole glycoalkaloid, with an overall purity of greater than 65%, and is entirely suitable for incorporation into synthetic diets. PMID- 10794184 TI - Efficient genetic analysis of fungal samples. AB - We present a method for rapid genetic analysis of small amounts of fungal material. Sterile glass slides, sufficiently small to fit in a standard PCR tube, were placed on agar inside a Petri dish. After a few days, fungal cultures start to overgrow the glass slides. Glass slides with attached mycelium were harvested, analysed microscopically, and placed into a standard PCR tube. Conserved primers flanking the ITS regions of rDNA repeat were used in a direct PCR with the fungal material. Sequence data were generated to be included in phylogenetic analyses to investigate the relationships of the studied mycorrhizal fungi from orchids. The mycelium attached to glass slides was also used for an in situ hybridization experiment using fluorescent labelled oligonucleotides as probes. Fluorescent signal was found throughout the cytoplasm when a probe specific to a site in the nuclear small subunit rRNA is used. PMID- 10794185 TI - Preparation and characterization of milk calcium salts by using casein phosphopeptide. AB - Milk calcium salt solution was prepared by the following procedures using casein phosphopeptides (CPP). To CPP solution, 1 M citric acid, 1 M CaCl2 and 1 M K2HPO4 were added with stirring, while adjusting the pH to 6.7. The prepared solution was left for 12 hr at 25 degrees C and then used for subsequent experiments, or lyophilized. The concentrations of organic phosphate of CPP, calcium, inorganic phosphate, and citrate in the typical milk salt solution were 7, 30, 22, and 10 mM, respectively, which were close to those in bovine milk. The lyophilized sample was easily dissolved in water. No crystal structure of hydroxyapatite was shown in the lyophilized milk calcium salts by X-ray diffraction analysis, although the pattern of KCl crystal was observed. The X-ray diffraction pattern of commercial whey mineral, which was prepared by precipitation at alkaline pH from rennet whey, was similar to that of hydroxyapatite. It was confirmed by high performance gel chromatographic analysis that the form of calcium phosphate in the milk calcium salts was similar to that of casein micelles. PMID- 10794186 TI - Domain-specific knowledge as the "dark matter" of adult intelligence: Gf/Gc, personality and interest correlates. AB - An enduring controversy in intelligence theory and assessment, the argument that middle-aged adults are, on average, less intelligent than young adults, is addressed in this study. A sample of 228 educated adults between ages 21 and 62 years was given an array of tests that focused on a broad assessment of intelligence-as-knowledge, traditional estimates of fluid intelligence (Gf) and crystallized intelligence (Gc), personality, and interests. The results indicate that middle-aged adults are more knowledgeable in many domains, compared with younger adults. A coherent pattern of ability, personality, and interest relations is found. The results are consistent with a developmental perspective of intelligence that includes both traditional ability and non-ability determinants of intelligence during adulthood. A reassessment of the nature of intelligence in adulthood is provided, in the context of a lifelong learning and investment model, called PPIK, for intelligence-as-Process, Personality, Interests, and intelligence-as-Knowledge (Ackerman, 1996). PMID- 10794187 TI - Effects of aging on automatic and effortful processes in bimanual coordination. AB - Two experiments are reported that compared younger and older adults on their performance of two bimanual temporal coordination tasks at varying movement speeds. In many cases, older adults performed as well as younger adults at all speeds of an in-phase coordination pattern and at slow speeds of an anti-phase pattern for both coordination accuracy and stability. Age differences tended to emerge most prominently at high speeds for the anti-phase pattern. These findings are consistent with the aging literature regarding automatic and effortful processing distinctions, suggesting that relative age differences become magnified when effortful resources are required for motor performance. PMID- 10794188 TI - "We had a nice little chat": age and generational differences in mothers' and daughters' descriptions of enjoyable visits. AB - Mother-daughter relationships are generally characterized by frequent contact and high levels of regard. Yet little is known about what mothers and daughters find enjoyable about their contact. Forty-eight pairs of older mothers (M = 76 years) and their middle-aged daughters (M = 46 years) and 44 pairs of middle-aged mothers (M = 47 years) and their young adult daughters (M = 21 years) (N = 182) described recent enjoyable visits. Younger mothers and daughters emphasized the daughters' entry into adulthood and the strength of their ties as agreeable, whereas older mothers and daughters focused on the larger kin network in describing their enjoyable visits. Some older mothers and daughters also made negative comments, suggesting greater individuation and less idealization of the relationship. Mothers in both age groups appeared to be more invested in the relationships than were their daughters. Implications for the strength of the mother-daughter tie are discussed. PMID- 10794189 TI - Positive affect and function as influences on self-assessments of health: expanding our view beyond illness and disability. AB - Longitudinal data from 851 elderly residents of a retirement community (mean age = 73 years) were used to examine the correlates of self-assessments of health (SAH) and the predictors of changes in SAH over several follow-up periods ranging from 1 to 5 years. The authors hypothesized that indicators of positive health, including feelings of energy and positive mood, social support, and active functioning, are as important in determining current and future SAH as negative indicators such as disease history, disability, medication, and negative mood. Results of cross-sectional and longitudinal analyses showed that functional ability, medication use, and negative affect were salient to people judging their health, but positive indicators of activity and mood had an even stronger, independent effect. These findings show the importance of attending to the full illness-wellness continuum in studying people's perceptions of health. PMID- 10794190 TI - Quality of life in assisted living homes: a multidimensional analysis. AB - This study examined the impact of four domains upon the quality of life (QOL) of senior residents living in assisted living homes: (a) demographic characteristics and health status, (b) social involvement, (c) facility characteristics, and (d) the social climate. Participants were 201 residents with functional impairments living in 55 different assisted living facilities in California. QOL was measured with three scales of depression, life satisfaction, and facility satisfaction. Bivariate correlations and ANOVAs found significant relations between at least one of the QOL measures and age, health status, social and family involvement measures, facility characteristics, and social climate measures. Social climate measures of cohesion, conflict, and independence had the strongest zero-order correlations. Regression analyses for the three QOL measures found cohesion to be the strongest predictor in all three regressions. Other QOL predictors in the regression analyses were fewer health conditions, participation in social activities, monthly family contact, and an environment low in conflict. Findings suggest that assisted living homes can improve resident QOL by creating a cohesive social environment, and encouraging social participation and family involvement. PMID- 10794191 TI - Cognitive function and apolipoprotein E in very old adults: findings from the Nun Study. AB - OBJECTIVES: The epsilon4 allele of apolipoprotein E (APOE) has been associated with Alzheimer' s disease and with milder forms of cognitive impairment. We investigated the possibility that the absence of the epsilon4 allele may predict the maintenance of high cognitive function among very old individuals. METHODS: Our data are from the Nun Study, a longitudinal study of aging and Alzheimer's disease in 678 Catholic sisters. All sisters participate in annual functional exams that include the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) battery of cognitive tests. High cognitive function was defined as intact scores on five of the CERAD tests. A total of 241 participants aged 75 to 98 met this criterion at the first exam. RESULTS: Findings showed that 62% of the 241 participants maintained intact scores on the five CERAD tests throughout their participation in the study. Life table analyses indicated that those without the APOE epsilon4 allele spent more time with intact cognitive function than those with the epsilon4 allele (p = .007). Cox regression analyses indicated that those without the epsilon4 allele had half the risk of losing their intact status during the study when compared with those with the epsilon4 allele (p < .01). DISCUSSION: Our findings suggest that the APOE epsilon4 allele may be included among the variables that predict high cognitive function in cognitively intact, very old adults. Although the presence or absence of the epsilon4 allele is known to be related to the risk of dementia, it also appears to be related to maintaining high levels of cognitive function in old age. PMID- 10794192 TI - Racial patterns in disabled elderly persons' use of medical services. AB - OBJECTIVE: This study examines the extent that older persons experience patterns of health service use that vary by race. METHODS: Using the 1989 NLTCS database, researchers estimate ten binomial logistic regressions of community medical service use by disabled White and Black older persons. Chow tests and Oaxaca decomposition analysis inform why racial differences continue to exist, although most elderly persons have Medicare. RESULTS: With similar medical conditions, Blacks are less likely to use services, particularly prescription medications and physician services. Use of some medical services is more likely for elderly Black persons who live in rural areas, small cities, and Western states, or who have more IADLs, joint and breathing problems, and broken parts. An Oaxaca decomposition indicates that differences in personal characteristics (e.g., income and health) do not fully explain racial differences in use of prescriptions and physician services. DISCUSSION: For reasons that are unrelated to financial assets, blacks remain vulnerable in their ability to access services commonly used by older persons. To remedy racial disparities in medical utilization, public policy must expand its focuses beyond health finance issues and consider differences in availability, accessibility, and acceptability. PMID- 10794193 TI - An assessment of the wealth holdings of recent widows. AB - OBJECTIVES: This study examines the wealth holdings of recent widows, how they compare to those of married couple households, and the potential to use this wealth to augment the incomes of widowed women. METHODS: Data from the Survey of Income and Program Participation are used to investigate the amount and composition of wealth held by four different groups: always married women, about to-be-widowed women, recent widows, and long-standing widows. Regression analyses assess the impact of group membership on wealth holding controlling for other sociodemographic factors, and annuity calculations assess the potential for wealth to augment income. RESULTS: About-to-be widowed women have fewer assets than intact couples, and there is a further decline in assets at the time of the husbands' deaths and in the ensuing period. Estimates of the annuity value of widows' wealth show that its liquidation would do little to improve the economic situation of the poorest widows. DISCUSSION: These findings parallel what is known about income changes that surround the death of a spouse. They also point to the need for additional research on the relationship between wealth holdings and mortality as well as the roles that health care costs, life insurance, and bequests may play in altering widows' wealth. PMID- 10794194 TI - Age differences in voluntary association memberships: fact or artifact. AB - OBJECTIVES: To determine if the curvilinear relationship between age and voluntary association memberships is due to compositional differences between age groups. METHODS: Data are drawn from 12 nationally representative surveys covering the period 1974-1994 and including items on membership in 16 types of associations. Age is categorized into 14 groups ranging from 18-24 through 85+. Patterns of age differences in number of association memberships are examined before and after controlling for several variables related both to age and to memberships. RESULTS: At the bivariate level, number of memberships increases through ages 40-44 and then declines fairly steadily. When controls for compositional differences are introduced, the curvilinear pattern of age differences is replaced by one showing an increasing number of memberships through ages 55-59, a slight decrease in the 60-64 age group, and then generally stable membership levels through the 85+ age group. DISCUSSION: The lower levels of voluntary association memberships of older persons consistently identified in bivariate analyses appear to be largely due to differences in compositional characteristics between the older and other age groups. PMID- 10794195 TI - Prevalence and correlates of depressive symptoms among Hispanic elders in Massachusetts. AB - BACKGROUND: Few studies have addressed depression among Puerto Rican and Dominican elders. This study examined the prevalence of depressive symptomatology and associated situational factors, among a representative sample of Hispanic elders in the state of Massachusetts and a neighborhood comparison group of non Hispanic whites (NHWs). METHODS: Participants included 715 Hispanic elders (429 Puerto Rican, 128 Dominican, and 149 other Hispanic) in Massachusetts and 238 NHW elders living in the same neighborhoods. Depressive symptomatology was measured with the Center for Epidemiologic Studies Depression Scale (CES-D). The associations between socioeconomic status, household arrangements, acculturation, health problems, and depression score were examined with multiple linear and logistic regression. RESULTS: Puerto Rican and Dominican elders had significantly greater prevalences of high depression scores when compared with neighborhood NHWs. After controlling for various social, demographic, and health characteristics, the scores of Puerto Ricans remained significantly greater than those of the NHWs, but not "Other Hispanic" groups. Higher CES-D scores were significantly associated with being female, living alone, and having a higher number of health problems, but not with income. Effects of acculturation were only found among Dominican elders. CONCLUSIONS: Puerto Rican elders experience high rates of depressive symptomatology that are associated with, but not fully explained by, high numbers of chronic health conditions. Further investigation is needed to both understand and treat this prevalent problem. PMID- 10794197 TI - Otogenic intracranial inflammations: role of magnetic resonance imaging. AB - The clinical course of acute otitis media is usually short, and the process terminates because of the host's immune system, the infection-resistant properties of the mucosal linings, and the susceptibility of the major organisms (beta-hemolytic streptococcus or pneumococcus) to penicillin. However, a small proportion (1% to 5%) of untreated or inadequately treated patients may experience complications. Prior to the development of an intracranial complication of otomastoiditis, warning symptoms or signs may be evident; these include severe earache, severe headache, vertigo, chills and fever, and meningeal symptoms and signs. Increasing headache, particularly temporoparietal headache near the affected ear, often indicates an impending intracranial complication. This symptom, often the only indication of an epidural abscess, demands prompt investigation and medical and surgical intervention. In our experience, computed tomography (CT) permits accurate diagnosis of acute coalescent or latent (masked) mastoiditis and its associated complications. However, magnetic resonance imaging (MRI) remains the study of choice to evaluate otogenic intracranial complications. This article demonstrates the important role of MRI in diagnosing various stages of acute otomastoiditis and its associated complications. PMID- 10794196 TI - Changes in functional health status of older women with heart disease: evaluation of a program based on self-regulation. AB - OBJECTIVES: This study involving 570 women aged 60 years or older with heart disease, assessed the effects of a disease management program on physical functioning, symptom experience, and psychosocial status. METHODS: Women were randomly assigned to control or program groups. Six to eight women met weekly with a health educator and peer leader over 4 weeks to learn self-regulation skills with physical activity as the focus. Evaluative data were collected through telephone interviews, physical assessments, and medical records at baseline and 4 and 12 months post baseline. RESULTS: At 12 months, compared with controls, program women were less symptomatic (p < .01), scored better on the physical dimension of the Sickness Impact Profile (SIP; p < 0.05), had improved ambulation as measured by the 6-minute walk (p < 0.01), and lost more body weight (p < .001). No differences related to psychosocial factors as measured by the SIP were noted. CONCLUSION: A self-regulation-based program that was provided to older women with heart disease and that focused on physical activity and disease management problems salient to them, improved their physical functioning and symptom experience. Psychosocial benefit was not evident and may be a result of measurement error or due to insufficient program time spent on psychosocial aspects of functioning. PMID- 10794198 TI - Congenital cholesteatoma and cholesterol granuloma of the temporal bone: role of magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) can be of significant benefit in characterizing expansile lesions of the petrous apex. MRI can be of particular help in distinguishing congenital cholesteatoma from cholesterol granuloma. Furthermore, with the advent of faster high-resolution scanning techniques, MRI can define precise spatial relationships of these masses with the middle and inner ear structures, internal carotid artery, jugular vein, and other structures of the skull base. MRI scanning is sensitive in detecting intracranial extension of these masses. MR angiography can provide additional information regarding the relationship of these masses with adjacent vascular structures and confirm patency of the adjacent vessels. In the postoperative period, MR scanning can help evaluate for complete removal, complication, recurrence, or formation of complicating granulation tissue. PMID- 10794199 TI - Cerebellopontine angle tumors: role of magnetic resonance imaging. AB - The cerebellopontine (CP) angle is bound anterolaterally by the posterior aspect of the petrous temporal bone and posteromedially by the cerebellum and pons. It contains important vascular structures and cranial nerves and is subject to a certain gamut of lesions, notably tumors with interesting radiological manifestations. Radiological investigation of these lesions has seen significant improvement in recent decades. Magnetic resonance is the imaging modality of choice for lesions of the CP angle and internal auditory canal. Lesions of the CP angles usually are divided into those native to the angle (vestibular schwannoma, meningioma, epidermoid, arachnoid cyst, metastases, lipoma, etc.) and those extending to the angle from adjacent structures (gliomas, ependymomas, choroid plexus papillomas, vascular malformations). Vestibular schwannomas are by far the most important lesion of the CP angle. PMID- 10794200 TI - Paraganglioma of the temporal bone: role of magnetic resonance imaging versus computed tomography. AB - Paragangliomas, also known as glomus tumors or chemodectomas, are tumors arising from chemoreceptor tissue (paraganglia), which are neural crest in origin and found in higher concentration along the glossopharyngeal and vagal cranial nerve. Three types of paragangliomas are related with the temporal bone: glomus tympanicum, glomus jugulare, and glomus vagale. The role of computed tomography (CT) and magnetic resonance imaging (MRI) in diagnosing these types of tumors is discussed, along with the importance of arteriography in aiding the diagnosis and treatment. This article discusses the choice between CT and MR based on clinical symptoms and tumor location, and illustrates the newest CT, MR, and angiography applications. A brief discussion on treatment options is given. PMID- 10794201 TI - Role of magnetic resonance imaging and diagnostic and interventional angiography in vascular and neoplastic diseases of the skull base associated with vestibulocochlear symptoms. AB - There are many vascular and neoplastic diseases as well as normal variants that produce the vestibulocochlear symptoms of pulsatile tinnitus, hearing loss, dizziness, and ataxia. Magnetic resonance imaging may be diagnostic, and magnetic resonance angiography/magnetic resonance venography have added to the ability of magnetic resonance to image vascular abnormalities. The extent of neoplasms is accurately assessed and complication of vascular lesions are clearly seen. However, detailed vascular anatomy requires high-quality selective angiography. This enables optimal treatment planning. Endovascular therapeutic intervention has a major role to play in conjunction with surgery of skull base lesions and may be curative in certain conditions, avoiding major surgical procedures. The interventionalist, however, must have an excellent knowledge of the external carotid circulation and all of its potential communications with the internal circulation to avoid serious embolic complications. PMID- 10794202 TI - Magnetic resonance imaging of the temporomandibular joint. AB - The spectrum of disease that affects the temporomandibular joint (TMJ) can be varied. To differentiate among the diseases that cause pain and dysfunction, an intimate knowledge of the anatomy, physiology, and pathology of this region is necessary. Due to the joint's complex anatomy and relationship to the skin, it has been difficult to image in the past. Magnetic resonance imaging is ideally suited for visualizing TMJ because of its superb contrast resolution when imaging soft tissues. Magnetic resonance imaging allows simultaneous bilateral visualization of both joints. The ability to noninvasively resolve anatomic detail can be performed easily and quickly using magnetic resonance imaging. The development of magnetic resonance imaging has greatly aided the diagnosis of TMJ disorders. An understanding of TMJ anatomy and pathogenesis of TMJ pain is crucial for interpretation of magnetic resonance imaging and subsequent treatment. PMID- 10794203 TI - Is Argentina ready for managed care? AB - The author describes the evolution and present status of health insurance and prepaid medical care in Argentina, with particular reference to the opportunities for the development of managed care systems and techniques there. He claims that as US health care corporations enter that country's marketplace there are lessons to be learned that will probably be applicable in other countries of the Americas. PMID- 10794204 TI - Decision-making in HIV prevention community planning: an integrative review. AB - Since 1994, the Centers for Disease Control and Prevention has required that the 65 health department grantees that receive funding for HIV prevention interventions engage in a community planning process to involve affected communities in local prevention decision making; to increase the use of epidemiological data to target HIV prevention resources; and to ensure that the planning process takes into account scientific information on the effectiveness and efficiency of different HIV interventions. Local community planning groups are charged with identifying and prioritizing unmet HIV prevention needs in their communities, as well as prioritizing prevention interventions designed to address these needs. Their recommendations, in turn, form the basis for the local health department's request for HIV prevention funding from the Centers for Disease Control and Prevention. Given the community planning process's central role in the allocation of federal HIV prevention funds, it is critical that sound decision-making procedures inform this process. In this article, we review the basics of the community planning prioritization process and summarize the decision-making experiences of community planning groups across the US. We then describe several priority-setting tools and decision analytic models that have been developed to assist in HIV community planning prioritization and discuss their strengths and weaknesses. Finally, we offer suggestions for improving the decision-analytic basis for HIV prevention community planning. PMID- 10794205 TI - Evaluation of a pilot program in rural schools to increase bicycle and motor vehicle safety. AB - Concerned with high bicycle-and motor vehicle-related mortality rates among children, Idaho's South Central District Health Department provided a competition to increase use of 1) seat belts, 2) motor vehicle rear seating, and 3) bicycle helmets among children attending elementary schools in the eight rural counties it serves. Nine of the 44 elementary schools in the health district chose to participate in the contest. Eight schools addressed increasing helmet use, four schools also addressed increasing seat belt use and rear seating, and one school addressed safety in general. A $1,000 prize was awarded to each of four schools judged to have the highest levels of student and community involvement, outreach, creativity, and changes in safety behavior (based on perceptions of outside judges). In 1997, baseline observations were collected for 1) seat belt use and rear seating for children in 28 schools, and for 2) bicycle helmet use among children in 25 schools. In 1998, follow up data were collected for 1) seat belt and rear seating in 42 schools, and 2) bicycle helmet use in 35 schools. Data were analyzed using SAS. Adjusting for differences in baseline rates, regression analysis was used to compare 1997 and 1998 rates for seat belt use, rear seating, and bicycle helmet use for those schools having baseline data. Results showed that although there was no significant difference between participating and non participating schools in rear-seating behaviors, there was an increase in seat belt and bicycle helmet use for participating schools. Since schools self selected participation, it is unknown whether those schools were fundamentally different from nonparticipating schools. PMID- 10794206 TI - Health-related predictors of self-perceived health in a student population: the importance of physical activity. AB - The main objective of the present study was to observe and describe the intercorrelations of a number of psychological and health-related variables with special emphasis put on the predictors of self-perception of health using stepwise linear regression analysis. Data were collected in conjunction with a research project of health risk behavior of the student population of Szeged, Hungary. The number of students participating in our sample was 980, the response rate was 70.5% (n=691). A self-administered questionnaire was used as a method of data collection. The findings suggest that psychological well-being plays a central role in determining self-perceived health. The regression analysis revealed that four health-related variables under study contributed significantly to the self-perception of health. These were the following: psychological well being, physical activity behavior, acute illness episodes and the frequency of psychosomatic symptoms. While some differences were detected between the type and the number of contributing factors among the subgroups by sex and physical activity behavior (i.e., physical activity was far more influential among males and likewise among more active persons) psychological well-being proved to be the strongest predictor. Physical activity behavior correlated positively and the frequency of psychosomatic symptoms negatively with psychological well-being. Moreover the occurrence of harmful habits was positively related both to the frequency of symptoms and physical activity behavior. PMID- 10794207 TI - Public policy involvement by nurse practitioners. AB - The purpose of this study was to determine the level and type of public policy involvement, as well as perceptions regarding public policy involvement of nurse practitioners. A four-page survey was mailed to a sample of 600 certified nurse practitioners, randomly selected from the American Academy of Nurse Practitioners' database. A three-stage mailing procedure yielded a 73% response rate. Most (59.6%) were involved in three or less public policy activities. The most frequently indicated activities included voting (87%) and giving money to a campaign (57%). Lack of time was the most frequently cited barrier, while improving the health of the public was cited most often as a benefit. Overall, nurse practitioners felt they had limited knowledge on how to go about changing public policy, were somewhat interested in public policy issues, believed the actions of public policymakers were very important, and believed these actions influenced the public's health. The majority (79%) had received some information/education on public policy change. Those most active in public policy had high public policy efficacy expectations and perceived a high number of benefits to public policy involvement. PMID- 10794208 TI - Uncontrollable frenzy and a unique temporary insanity plea. AB - On a bright, sunny April 1859 afternoon in Washington, D.C., a brutal murder occurred in Lafayette Park, directly across from the White House. Ironically, a close friend of President James Buchanan shot and killed his wife's lover. Daniel E. Sickles, a cuckolded U.S. Congressman, attacked and killed Philip Barton Key, the U.S. Attorney for the District of Columbia. Key was one of the eleven children of Francis Scott Key, the author of the poem that became The Star Spangled Banner. At the trial, Sickles' seven lawyers presented an insanity plea claiming that an "uncontrollable frenzy" created a 'brainstorm" resulting in temporary insanity. In addition, the defense argued that the "unwritten law" provided immunity to a husband who avenged his honor. Only lay witnesses testified as to Sickles' intense emotional outrage. There was no expert medical witness to support the insanity plea. Prosecutors maintained that the killing was a premeditated murder, or at the very least manslaughter. PMID- 10794209 TI - Laparoscopic total extraperitoneal hernia repair: mesh fixation is unnecessary. AB - BACKGROUND: Inguinal hernia repair contributes significantly to the general surgeon's workload. Since the evolution of laparoscopic inguinal hernia repair, the total extraperitoneal (TEP) repair is the technique most commonly employed by laparoscopic surgeons. This technique involves the placement of a polypropylene mesh in the preperitoneal space. The issue of fixation of this mesh remains unresolved. Surgeons have previously fixed this mesh in place using laparoscopic stapling devices, suturing techniques, or, more recently, polycyanoacrylate adhesives. However, stapling the mesh not only increases the time and expense of the procedure but can cause specific complications such as nerve entrapment syndromes and osteitis pubis. PATIENTS AND METHODS: We report a series of 89 total extraperitoneal laparoscopic repairs in 80 consecutive patients using no means of mechanical or adhesive mesh fixation, irrespective of the size of the hernial defect. RESULTS: Follow-up revealed no increase in morbidity or hernia recurrence. CONCLUSION: Our experience suggests that mechanically fixing the mesh in the preperitoneal space is unnecessary. Not fixing the mesh avoids possible complications and is not associated with any increased risk of hernia recurrence. PMID- 10794210 TI - Physicians' choice for their own hernia repairs. AB - BACKGROUND AND PURPOSE: While the optimal method of inguinal herniorrhaphy is controversial, there is growing acceptance that laparoscopic hernia repair is a legitimate alternative to conventional techniques. This study sought to determine if physicians as patients had different preferences for their own hernia repairs than nonphysician patients. PATIENTS AND METHODS: Total endoscopic preperitoneal (TEP) herniorrhaphy was introduced into the author's practice in 1995. Open herniorrhaphies (OH) were performed under local anesthesia and were almost all tension-free repairs. Patients were given the option of surgical technique after a discussion with the author, although patients with primary unilateral hernias were encouraged to undergo a tension-free OH. A prospective database was kept and subsequently analyzed. RESULTS: In the 3 years from June 1, 1995, to June 1, 1998, a total of 138 OH and 77 TEP repairs were performed. There were 19 physicians among the 215 patients. During the 3-year period, the annual percentage of laparoscopic herniorrhaphies increased from 27% (21/79) to 46% (32/70) (P = 0.024). The shift in physician preference for TEP from 16% (1/6) in 1995 to 75% (6/8) in 1997 was more dramatic than the shift in the population at large: 22% (20/73) to 42% (26/62). All patients undergoing TEP repair for recurrent hernias stated their recovery was easier than after their original OH. Four of seven physicians with recurrent hernias also had bilateral hernias. None required hospitalization. The median time to return to work was 4 days in the TEP physician group and 7 days in the physician OH group. The median time to return to work was 10 days in the TEP nonphysician group and 16 days in the OH nonphysician group. CONCLUSIONS: Physicians cared for by the author are increasingly choosing a laparoscopic approach for their hernia repairs even when they have primary unilateral hernias. Patients return to work more rapidly after TEP repairs than after OH. PMID- 10794211 TI - Laparoscopic ventral hernia repair. AB - PURPOSE: A retrospective study was carried out of patients who underwent laparoscopic ventral abdominal wall hernia repair (excluding groin hernias) between January 1994 and January 1999. PATIENTS AND METHODS: Laparoscopic ventral hernia repair was performed on 202 patients for defects ranging from 1.5 cm to 12 cm in diameter. Of these, 35 patients had multiple hernial defects. After reduction of the hernial contents and adhesiolysis, a polypropylene mesh was used intraperitoneally in all patients, such that there was a margin of at least 3 cm from the edge of the defect as well as the previous scar. RESULTS: The mean operating time decreased from 90 minutes in the initial 3 years to 50 minutes in the last 2 years. Postoperatively, the mean hospital stay was 1.8 days. Patients complained of somatic abdominal pain at the site of mesh insertion for a mean of 7 days. There were two postoperative hernia recurrences at a mean follow-up of 2.9 years. The incidence of seroma formation postoperatively was 32% in the first 3 years but declined to 18% subsequently with postoperative abdominal-wall pressure dressings. There were no postoperative sequelae related to bowel adhesions. Negligible wound sepsis (superficial wound infection in five patients), decreased morbidity, and all the other advantages of a minimally invasive surgical approach were evident in this group of patients. CONCLUSION: These promising early results need to be confirmed by a prospective controlled trial, especially recurrence rates and incidence of postoperative adhesions. PMID- 10794212 TI - Is balloon ablation as effective as endometrial electroresection in the treatment of menorrhagia? AB - BACKGROUND: Hot-fluid balloon therapy is a recently introduced, relatively simple endometrial ablation procedure for menorrhagia. Because it is thought to be safer than other ablation procedures, it would be superior to other types of ablation if it is equally effective. The purpose of the present study was therefore to compare the safety and effectiveness of balloon ablation and transcervical resection of the endometrium (TCRE) for the treatment of menorrhagia. PATIENTS AND METHODS: We performed a prospective cohort study comparing TCRE and hot-fluid balloon ablation in consecutive patients suffering from menorrhagia and not responding to medical treatment. Between 1992 and 1994, all patients had TCRE, whereas from 1995 onward, all patients had balloon therapy. Outcome measures were surgical reintervention, menstrual pattern, and patient satisfaction. Assuming a 9% reintervention rate after TCRE, a series of 150 patients was required to show balloon ablation to be equally effective. RESULTS: Of the 152 patients who were included, 75 underwent TCRE and 77 had balloon ablation. The procedure had to be abandoned in 13 patients in the TCRE group (17%) and in 8 patients in the balloon ablation group (10%). In the TCRE group, four patients underwent a second resection, whereas hysterectomy was performed in 15 patients (3-year cumulative reintervention rate 26%). In the balloon-ablation group, there were no reresections, whereas hysterectomy was performed in 9 patients (3-year cumulative reintervention rate 13%) (log-rank test P = 0.11). The relative risk for any reintervention was 0.36 (95% confidence interval 0.05-2.5). At 3 months' follow up the duration of menstruation was significantly shorter after TCRE than after balloon ablation, but at 6, 12, and 24 months, the duration of menstruation in the two groups appeared to be equal. No difference in patient satisfaction could be detected between the two groups, but there appeared to be a statistically significant decline in patient satisfaction over time for both therapies. This decline was stronger after TCRE than after balloon ablation. CONCLUSIONS: Because endometrial ablation with a hot-fluid balloon seems to be as effective as endometrial resection, with a lower complication rate, balloon ablation might become the procedure of choice for endometrial ablation. PMID- 10794213 TI - Development and initial trial of the minilaparoscopic argon coagulator. AB - BACKGROUND: A major limitation of minilaparoscopic or needlescopic instrumentation is the lack of a proficient mechanism by which to effect coagulation and hemostasis. Our purpose was to test the feasibility of the first minilaparoscopic argon coagulator (MAC). Performance requirements insist on a system that ensures noncontact ignition and coagulation, a low argon gas flow rate, and safety and efficacy at a variety of wattages. MATERIALS AND METHOD: The first MAC was formulated and produced in the laboratory. Among its unique properties was that it contained a corona ignition system. The efficacy of the system was measured in the laboratory by advancing the prototype instrument, while under power and with the argon flowing, toward a brass screen connected to the electrosurgical generator return. The distances at which an arc formed at various wattages were measured via a micrometer translation stage and recorded as the ignition gap. After the successful laboratory trials, an experiment was conducted in four 30-kg pigs. A gastroscope was introduced for vision and insufflation, and transcutaneous, transgastric placement of needlescopic instruments and the MAC was performed. Areas of mucosa 1 x 2 cm were coagulated at 15, 25, 35 and 40 W in each animal. Ulcers of 1 cm were created and coagulated with the MAC. Two animals were sacrificed immediately and one each at 12 and 14 days, and the coagulated areas were examined histologically. RESULTS: In the laboratory, ignition of the beam was induced without surface contact, and the desired 5-mm ignition gap was seen at power as low as 15 W. The argon gas flow rates were limited to 1.5 L/min. At each wattage, the mucosa was entirely ablated. The depth of injury varied (submucosa to 2.2 mm) according to the wattage applied. Ulcerative hemorrhage was controlled in each instance. There were no complications attributable to coagulation effects. Surviving animals tolerated a regular diet on postoperative day 1. Histologic examination of tissue from surviving animals revealed growth of normal mucosa in the superficially coagulated areas and mixed normal mucosa and fibrosis in the more deeply injured regions. CONCLUSIONS: Both in the laboratory and in animal experiments, the first MAC demonstrated noncontact ignition and operation, a low flow rate, and safety and efficacy at all wattage settings applied. It arrested ulcerative bleeding and ablated the mucosal surface at various depths according to the applied power. The MAC may be a solution to the lack of a consistent and reliable means for coagulation in needlescopic or minilaparoscopic surgery. PMID- 10794214 TI - Retroperitoneoscopic lumbar sympathectomy with balloon dissection: clinical experience. AB - PURPOSE: To describe our technique and clinical experience of retroperitoneoscopic lumbar sympathectomy with balloon dissection. PATIENTS AND METHODS: From 1995 to 1999, 23 procedures were performed. RESULTS: No complications were noted. The median hospital stay was 4 days. Of the 23 attempts, 2 had to be converted to open technique. Histologic examination demonstrated interruption of the sympathetic chain in 22 of the 23 cases and absence of nervous tissue in 1. CONCLUSIONS: Once the technique is mastered, it can be performed fast, and it offers patients the benefits of a minimally invasive approach without the inconsistent therapeutic results of chemical sympathectomy. PMID- 10794215 TI - Laparoscopic repair of the leaking percutaneous endoscopic gastrostomy. AB - Peritonitis is an infrequent yet major complication of a percutaneous endoscopic gastrostomy (PEG). Traditionally, patients with peritonitis from leaking PEG tubes underwent open abdominal exploration with repair of the gastrostomy site. We report successful laparoscopic treatment of this significant complication. Surgical techniques and technical aspects of the procedure are discussed. PMID- 10794216 TI - Laparoscopic management of a late-diagnosed major diaphragmatic rupture. AB - A 20-year-old male patient was admitted to our emergency ward because of acute respiratory insufficiency following gastroscopy 2 years after a car accident. The chest radiograph showed migration of the stomach into the left hemithorax. A large diaphragmatic hernia was diagnosed and repaired laparoscopically using slowly resorbable sutures and patches. Diaphragmatic rupture secondary to blunt thoracic or abdominal trauma is a rare injury, whose diagnosis may be delayed. The majority of these defects are diagnosed during laparotomy performed for other major abdominal lesions. If diaphragmatic rupture is suspected, and no lesion of a parenchymatous organ has been diagnosed, there is a role for diagnostic laparoscopy. In the absence of other abdominal injuries, diaphragmatic rupture can be repaired by minimal-access surgery. PMID- 10794217 TI - Laparoscopic pancreatic necrosectomy. AB - We describe a patient with infected pancreatic necrosis who was treated successfully with minimally invasive surgery. Five weeks after an episode of acute uncomplicated pancreatitis, he was found to have infected pancreatic necrosis and splenic vein thrombosis. The patient underwent a laparoscopic pancreatic necrosectomy, splenectomy, and cholecystectomy. Seven days after surgery, the patient was discharged and continued to be asymptomatic for the 6 months of follow-up. PMID- 10794218 TI - Testing surgical gowns for the "anticipated level of exposure". AB - Although the use of the surgeon's gown dates back to the turn of the century, the need for it to be made of a liquid-repellent material was disclosed only in 1952. Because of the relatively poor performance of the products that were introduced early on, the entire textile industry--makers of nonwoven disposable and woven reusable materials alike--was challenged to develop a test method to demonstrate a fabric's capability "under usual conditions of use." A cooperative attempt to do that was abandoned in 1983. With the emergence of HIV, the need to protect the wearer became the gown's priority. However, because there was no standard test method, the manufacturers used any of an array of tests to promote a product's suitability for use under what the Occupational Safety and Health Administration describes as the "level of exposure anticipated." Now, a standard test method has been adopted that describes the results on a pass/fail basis. However, the literature indicates that gowns made of materials that have passed this test have failed "under usual conditions of use." Nevertheless, the Food and Drug Administration is permitting manufacturers to mislead the surgical community by describing products as being "impervious" or "liquid proof." PMID- 10794219 TI - LiteratureWatch. PMID- 10794220 TI - 20-year experience with cemented primary and conversion total hip arthroplasty using so-called second-generation cementing techniques in patients aged 50 years or younger. AB - We present the 20-year experience of 47 hips in 40 patients aged 50 years or younger with cemented primary total hip arthroplasty using second-generation femoral cementing techniques. Average follow-up duration in the 23 patients living at least 17 years was 18.2 years. Overall, 18 hips (38%) had components revised or removed for any reason, at an average duration of 12.6 years. Every revision or reoperation involved removing the acetabular component. Of these 18 acetabular components, 15 (32%) were revised for aseptic loosening. Eleven additional acetabular components were loose by radiographic criteria at final follow-up, yielding prevalence of aseptic acetabular loosening (55%). Four femoral components (8%) were revised for osteolysis without loosening, and 3 (6%) were revised for aseptic loosening. Femoral osteolysis, with or without component loosening, led to revision in 5 ( 11%) hips compared with 6% for aseptic loosening alone. Osteolysis was the primary problem leading to acetabular and femoral component revision in this series of people < or = 50 years old over the first 20 years after the index operation. PMID- 10794221 TI - The hydroxyapatite-ABG hip system: 5- to 7-year results from an international multicentre study. The International ABG Study Group. AB - A proximally hydroxyapatite (HA)-coated hip prosthesis designed for proximal stem bonding and stress transfer was studied in a consecutive series of 398 patients in an ongoing, prospective multicenter study. Follow-up was 5 to 7 years. The clinical results were excellent with a mean preoperative Merle d'Aubigne score of 8.5 and 17.3 at 5 years. Only mild thigh pain not necessitating analgesics or a walking stick was reported in 3.6% at 5 years. Three cups and 3 stems had to be revised, all before the 2-year assessment and mainly for technical reasons. No components needed revision after 2 years. The 5-year HA-coated component survival was 99.2%. Serial radiographs showed excellent osseointegration, which was independent of pre-existing bone stock. The rate of radiographic bone remodeling was highest during the first 3 years, and positive bone remodeling (bone formation) preceded negative bone remodeling (bone resorption). The concept of transitional load transfer from proximal to distal could be acknowledged, with proximal load transfer highest in 77%. The combination of anatomic stem design with distal overreaming and proximal HA coating was thought to be the reason for these excellent results. Proximal circumferential osseointegration seems to be real because no linear or distal osteolysis was observed. PMID- 10794222 TI - Computer-assisted decision analysis in orthopedics: resurfacing the patella in total knee arthroplasty as an example. AB - The purpose of the present study was to illustrate the use of computer-assisted decision analysis in making decisions in the field of orthopaedic surgery, using the choice between resurfacing and not resurfacing the patella in total knee arthroplasty as an example. We used a decision analysis technique based on probability theory and on Bayesian logic, with the help of an especially developed computer software. The process involves building a decision tree, searching for probabilities and utilities in the literature, folding back the tree to compute the baseline result, and running sensitivity analyses. Our literature search provided 26 useful articles, only 3 of which were randomized controlled trials. In the baseline analysis, both options were rated similarly, with resurfacing the patella faring slightly better. Sensitivity analyses revealed that not resurfacing becomes the procedure of choice if the probability of postoperative anterior knee pain with an unresurfaced patella falls below 14%, or if the probability of having pain with a resurfaced patella rises above 8% or if the utility of patellar implant failure falls below 80% of the utility of a perfect health state. Computer-assisted decision analysis is a promising, evidence-based tool to assist clinical decision making in orthopaedic surgery. However, its validity is limited by the poor quality of data found in the orthopaedic literature, especially the scarcity of randomized controlled trials. PMID- 10794223 TI - Balance sheets of knee and functional scores 5 years after total knee arthroplasty for osteoarthritis: a source for patient information. AB - To improve patient information on the results of total knee arthroplasty (TKA) for osteoarthritis, 253 primary TKAs of a prospective study with a mean follow-up of 5.3 years were analyzed. The increase or decrease of the individual variables of the Knee Society knee and function score and the percentage of operated knees in which these variables increased were determined. Improvement in pain rating had the largest increase of all variables, contributing 60% to the knee score increase. Pain improved in 95% of the knees. Alignment improved in about 90% of knees and accounted for 25% of knee score increase. Improvement in level walking contributed more to increase of function score than better stair-climbing abilities. Level walking improved in 80% of knees and stair climbing in 55%. Pain is the most rewarding indication for TKA, followed by deformity and poor walking ability. Key words: total knee arthroplasty, knee score, functional score, patient information. PMID- 10794224 TI - The predictive value of indium-111 leukocyte scans in the diagnosis of infected total hip, knee, or resection arthroplasties. AB - To evaluate the usefulness of the indium-111 scan in detecting actually or potentially infected total hip, knee, and resection arthroplasties, 153 scans were performed on 143 patients who underwent reoperation for a loose or painful total joint arthroplasty or a resection arthroplasty between 1990 and 1996. Scans were interpreted as infected, not infected, or equivocal by an experienced nuclear medicine radiologist. Patients were considered to be infected if they met any 2 of the following criteria: i) positive intraoperative cultures, ii) final permanent histologic section indicating acute inflammation, and iii) intraoperative findings of gross purulence within the joint. Twenty-six patients (17%) met the infection criteria at the time of reoperation. Indium scans were found to have a 77% sensitivity, 86% specificity, 54% and 95% positive and negative predictive values, and 84% accuracy for the prediction of infection. Of 6 equivocal scans, none were infected. The results of this study suggest limited indications for the use of the indium-111 scan in the evaluation of painful hip, knee, or resection arthroplasties. A negative indium scan may be helpful in suggesting the absence of infection in cases in which the diagnosis is not otherwise evident. PMID- 10794225 TI - White blood cell counts and differential in synovial fluid of aseptically failed total knee arthroplasty. AB - A cell count and differential was obtained on synovial fluid samples from 79 total knee arthroplasties undergoing revision for aseptic failure over a 5-year period. The mean white blood cell count (WBC) was 782/mL (range, 11-7,200/mL). The mean percentage of monocyte cells was 87%, and the mean polymorphonuclear leukocyte (PML) cell count was 13%. Eight patients (10.4%) had leukocyte cell counts >2,000. Five knees in 4 patients with rheumatoid arthritis had PML counts >50%. There was a single postoperative infection. In patients with osteoarthritis as a primary diagnosis, a synovial WBC of <2,000 and a differential with <50% PMLs had a 98% negative predictive value for the absence of infection. Key words: synovial fluid, cell count and differential, failed total knee arthroplasty, revision total knee arthroplasty. PMID- 10794226 TI - The spatial location of impingement in total hip arthroplasty. AB - Impingement between acetabular and femoral components produces wear debris and results in abnormal loads on the edge of the implant. To characterize further the spatial location of impingement and the design and alignment factors associated with impingement, we reviewed 111 retrieved acetabular components from a single manufacturer. The location of impingement in the pelvis was determined by combining the location of impingement in the retrieved implants and the spatial orientation of the acetabular components measured from available radiographs. Evidence of impingement was identified in 39% of the retrieved implants and involved the posterior portion of the acetabulum in all cases. Posterior impingement was probably the result of femoral extension and external rotation, a motion that occurs during the toe-off phase of the gait cycle. Cups with impingement were more anteverted than those without impingement (P = .016). There was a significant inverse association between impingement and the size of the femoral head, and the mean head-to-neck diameter ratio for implants with impingement was smaller than that for implants without impingement (P < .0001). Factors that appear to be associated with impingement include i) excessive cup anteversion combined with posterior positioning of the extended rim and ii) femoral components with relatively small head-to-neck diameter ratio. PMID- 10794227 TI - Size and direction of hip joint forces associated with various positions of the acetabulum. AB - When total hip replacement is performed, the position of the acetabular component may affect wear and component survival time. We considered the questions: In what way does displacement of the hip joint center alter (1) the magnitude and (2) the direction of the resultant force? Biomechanical tests were carried out on a human multibody model. After displacement of the hip joint center, the resultant forces were calculated for the single leg stance. With the flexed single leg stance, maximum hip joint forces were observed with lateral, cranial, posterior displacement. The peak forces were affected by the modeling of a gluteus maximus wrapping point at the ischial tuberosity and were overestimated when this was removed. With the straight single leg stance, posterior displacement decreases the total load on the hip joint because of the increased leverage of the rectus femoris. With regard to the direction of the resultant force, medial displacement increases the angles in both planes, cranial displacement increases it in the sagittal plane (cranial, posterior-caudal, anterior), and anterior displacement decreases the angle in the sagittal plane and increases it in the frontal plane (medial, cranial-lateral, caudal). The direction of the force is relatively insensitive to displacement of the hip joint center. The results presented here indicate a marked increase in the force after lateral, cranial, posterior displacement of the center in the flexed single leg stance. To avoid extreme joint loading and to reduce the wear after total hip arthroplasty, the cranial and posterior regions of the acetabulum should be fully reconstructed. A high hip joint center has an adverse effect on the magnitude of the force, although the directions are hardly affected by it. PMID- 10794228 TI - Influence of mechanical behavior on the wear of 4 clinically relevant polymeric biomaterials in a hip simulator. AB - The elastic and large-deformation mechanical behavior of 4 materials with known clinical performance was examined and correlated with the wear behavior in a hip simulator. Acetabular liners of a commercially available design were machined from ultra-high molecular weight polyethylene (UHMWPE), high-density polyethylene (HDPE), polytetrafluoroethylene (PTFE), and polyacetal and wear tested in a multidirectional hip joint simulator. Elastic and large-deformation mechanical behavior was directly measured from the wear-tested liners using the small punch test. The finite element method was used to compute elastic modulus from the measured small punch test initial stiffness, and the contact stress for the liners was calculated using the theory of elasticity solution. Positive, statistically significant correlations were observed between the hip simulator wear rate and the initial peak load, ultimate load, and work to failure from the small punch test. Negative correlations were observed between the wear rate and the elastic modulus and contact stress. The results of this study support the hypothesis that the large-deformation mechanical behavior of a polymer plays a greater role in the wear mechanisms prevalent in total hip replacements than the elastic behavior. PMID- 10794229 TI - Reduction of polyethylene wear by concave dimples on the frictional surface in artificial hip joints. AB - For the purpose of reduction of polyethylene wear generated in artificial hip joints, concave dimples were formed on the surface of the metal femoral head by electrical discharge etching with the constant diameter of 0.5 mm, pitch of 1.2 mm, and depth of 0.1 mm. Using a hip joint simulator, the total amount of polyethylene wear was 7.2 mg in the concave-patterned condition and 23.1 mg in the nonpatterned condition. The reasons for the reduction of polyethylene wear by the concave pattern may be i) a reduction of abrasive wear by providing escape dimples for wear particles and ii) an improvement in the lubricity on the frictional surface by the supply of lubricant that is stored in dimples. PMID- 10794231 TI - Anatomic and biomechanical aspects of pie crusting posterolateral structures for valgus deformity correction in total knee arthroplasty: a cadaveric study. AB - Correction of valgus deformity during total knee arthroplasty is usually carried out by releasing lateral supporting structures from the femoral side of the joint. A new technique has been advocated that involves multiple stabs of the scalpel blade or pie crusting of the posterolateral corner. It is the hypothesis of this study that the correction achieved by using this technique occurs when the lateral collateral ligament is effectively released and that the common peroneal nerve may be at risk. Using a cadaveric model with 6 knees tested, significant differences were determined between 2 separate pie crusting steps as well as between releasing the lateral collateral ligament and popliteus tendons. Anatomic dissection studies also showed that in full extension the peroneal nerve may be less than the depth of a number 11 blade (16 mm) from the posterolateral corner, and the nerve may be at risk during this technique. These results show that major deformity correction obtained using the pie crusting technique is probably through effective release of the lateral collateral ligament. PMID- 10794230 TI - Osteogenic protein 1 device stimulates bone healing to hydroxyapaptite-coated and titanium implants. AB - This study evaluated the effects of osteogenic protein 1 (OP-1) placed in a gap around uncoated and hydroxyapatite (HA)-coated implants. Unloaded cylindrical titanium alloy implants were inserted in the femoral condyles of 16 skeletally mature dogs surrounded by a 3-mm gap. The gap around the implants was filled with 325 microg OP-1 in 130 mg bovine collagen type I as carrier (OP-1 device) or collagen carrier alone or left empty. All groups were tested with both HA-coated and uncoated implants, and the animals were sacrificed after 6 weeks. Implant fixation was determined by push-out test. Bone ongrowth and bone formation were evaluated by quantitative histomorphometry. OP-1 device enhanced mechanical fixation of uncoated and HA-coated implants, resulting in a higher shear strength than implants with collagen matrix and control implants. Bone ingrowth and bone formation in the gap were also stimulated 3-fold for OP-1 groups when compared with empty controls, but no difference was found between OP-1 groups and collagen matrix groups. This study suggests that the OP-1 device placed in a gap around uncoated and HA-coated implants is capable of enhancing mechanical fixation and periimplant bone formation. The collagen carrier alone also enhanced bone ongrowth and fixation significantly. PMID- 10794232 TI - Anatomy of the epicondyles of the distal femur: MRI analysis of normal knees. AB - Knowledge of precise anatomic landmarks and relationships of the distal femur can be helpful in knee surgery, especially primary and revision total knee arthroplasty. We analyzed 104 consecutive routine knee magnetic resonance imaging studies to define useful landmarks and relationships. The epicondyles are described, and the relationship of the epicondyles to the joint line is defined in multiple planes. Some significant gender differences were noted. The distance from the epicondyles to the joint line correlates with the transepicondylar width of the distal femur. This information can be helpful in determining appropriate joint line position intraoperatively. The posterior condylar angle averaged 3.11 degrees for all patients, and a tendency for the posterior condylar angle to increase with age was noted, but further study of this tendency is needed. PMID- 10794233 TI - Negative pressure intrusion cementing technique for total knee arthroplasty. AB - Negative pressure intrusion (NPI) is an alternative cementing technique for the tibial baseplate of total knee arthroplasty that uses a suction cannula in the proximal tibia to remove excess fluids and fat before cementing. This technique was compared with standard third-generation positive pressure intrusion (PPI) techniques in an in vitro implantation and analysis of 6 pairs of cadaveric tibiae. Six matched pairs of fresh frozen tibiae were prepared by cutting the tibial surfaces, standard cleaning and surface drying, then performing PPI and NPI on 1 of each pair. No objective differences were noted on radiographs or direct cement depth measurement analysis. Scanning electron micrograph evaluation revealed that the PPI specimens had consistently more voids in the cement-bone composite, and the NPI specimens had consistently narrower empty spaces between bone and cement, resulting in tighter fill in NPI specimens. NPI was shown to enhance characteristics known to improve tensile and shear strength in cement bone composites. PMID- 10794234 TI - Ordering allograft by weight: suggestions for the efficient use of frozen bone graft for impaction grafting. AB - Impaction grafting of morcellized allograft gives good early results in revision arthroplasty in which there is aseptic endosteolysis. Most surgeons employing this technique use fresh frozen allograft from femoral heads harvested at primary arthroplasty and order bone from the tissue bank in terms of numbers of femoral heads. This article demonstrates that this approach results in great variability in the quantity of graft available for impaction because of the variability in size and density of femoral heads. This variability may compromise the stability of the impacted graft. Surgeons in Raigmore Hospital request allograft by weight not quantity, which predicts more accurately the volume of graft after impaction. Having reviewed the figures to date, we suggest the amount of graft that should be available for certain procedures. PMID- 10794235 TI - Comparison of head penetration into Hylamer and Enduron polyethylene liners: a follow-up report. PMID- 10794236 TI - Acute cauda equina syndrome after total knee arthroplasty as a result of epidural anesthesia and spinal stenosis. AB - An unusual complication of epidural anesthesia performed for routine total knee arthroplasty is presented. Epidural catheter placement or removal resulted in an acute cauda equina syndrome in a patient with asymptomatic high-grade lumbar spinal stenosis at L3-4. The case is presented along with a literature review. PMID- 10794237 TI - Total knee arthroplasty for chronic posterior knee dislocation: report of 2 cases with technical considerations. AB - This article reports our experience in 2 patients with chronic knee dislocations treated with total knee arthroplasty, focusing on the technical difficulties in obtaining a well-balanced, well-aligned knee. PMID- 10794238 TI - Early catastrophic failure of rotating hinge total knee prosthesis. AB - The mechanical failure of a prosthesis component is usually observed as a late complication of total knee arthroplasty (TKA). In knees with severe ligament instability and bone deficiency, either a true hinge prosthesis or a rotating hinge implant is commonly used. Failure of the polyethylene bearing bush in a hinge-type prosthesis is a complication that has not been reported to date. We report the cases of 2 rotating hinge TKA prostheses that dislocated as a result of mechanical failure of the prosthetic component within 5 months of initial implantation. Clinicians should be aware of this potential complication when selecting rotating hinge prostheses for certain patients. PMID- 10794239 TI - Insidious destruction of the hip by Mycobacterium tuberculosis and why early diagnosis is critical. AB - Tuberculosis has re-emerged as an important problem in the United States. More than 10 million people presently are infected with Mycobacterium tuberculosis in the United States alone. The symptoms at first presentation of the disease have become more diverse. With extrapulmonary manifestations, such as musculoskeletal infections, as the sole presenting sign, it often can be difficult to determine the correct diagnosis early in the course of the disease. The presenting symptoms, physical signs, and radiographic findings of intra-articular tuberculosis can mimic those of other intra-articular diseases, such as rheumatoid arthritis, osteoarthritis, and avascular necrosis. In view of the nonspecific findings early in course of the disease, tubercular infection should be considered in the differential diagnosis when there is insidious articular destruction. Failure to consider tuberculosis can lead to devastating outcomes otherwise preventable with today's chemotherapies. PMID- 10794240 TI - Failure after customized curved femoral stems in total hip arthroplasty for Paget's disease. AB - We describe 2 patients with Paget's disease who underwent total hip arthroplasty for osteoarthrosis. In view of the femoral deformity in each case, a custom-made, long, curved femoral stem was used. Both patients have subsequently sustained periprosthetic fractures at the level of the tip of the prosthesis. PMID- 10794241 TI - Reducing spray at the time of joint replacement. PMID- 10794242 TI - Cyclosporine treatment of RPE allografts in the rabbit subretinal space. AB - PURPOSE: To determine the effects of systemic cyclosporine A (CsA) on the survival of retinal pigment epithelial (RPE) allografts in the subretinal space in an animal model using atraumatic transplantation surgery. METHODS: Following pars plana vitrectomy, an RPE cell suspension from brown rabbits was injected with a glass micropipette into the subretinal space of 39 albino rabbits. For immunosuppression, 22 rabbits were given an injection of CsA, 20 mg daily intramuscularly, 17 rabbits with RPE grafts were controls. The grafts were monitored by biomicroscopy, color fundus photography, and fluorescein angiography. Rabbits were sacrificed at 1, 3 and 6 months, respectively, and the eyes processed for light and electron microscopy including immunohistochemistry. RESULTS: After three months, the transplanted RPE cells, in both the CsA group and the controls, formed a monolayer in the subretinal space. Although a few macrophages were encountered, there was no massive cellular infiltration and the photoreceptor layer was well preserved. After six months, however, there was a disruption of grafted RPE cells in both groups, characterized by dispersion of melanin pigment in the subretinal space, and invasion of macrophages with focal photoreceptor damage but no infiltration of lymphocytes in the retina or choroid. No significant differences between the CsA treated and the control eyes were discernible. CONCLUSION: Although the subretinal space has been considered an immunologically privileged site, we found that the survival of RPE allografts was limited. CsA did not prevent RPE allograft destruction in the subretinal space. The transplant seems to be disrupted either by immunological mechanisms that are not inhibited by CsA, or by nonimmunologic events. PMID- 10794243 TI - Immortalization of human corneal endothelial cells using electroporation protocol optimized for human corneal endothelial and human retinal pigment epithelial cells. AB - PURPOSE: In this study we established a protocol for transfection of human corneal endothelial and human retinal pigment epithelial cells. This protocol was used for immortalization of human corneal endothelial cells. METHODS: Transfection was performed by means of electroporation. For immortalization a plasmid encoding large and small SV40 T-antigen was used. RESULTS: The established electroporation protocol was suitable for both cell types. This protocol was used for transfection of human corneal endothelial cells with a plasmid containing the early region of SV40. The transfected cultures exhibited an increased life-span before they entered crisis. One culture recovered from crisis and was cultivated for 300 population doublings. The cells exhibited an in vivo-like morphology usually lost during cell culture. CONCLUSIONS: We describe for the first time a culture of SV40 transfected human corneal endothelial cells which recovered from crisis and can therefore be regarded as immortalized. PMID- 10794245 TI - A new induction-based impact method for measuring intraocular pressure. AB - PURPOSE: An improved induction-based impact method for measuring intraocular pressure is presented. METHODS: A light probe containing a permanent magnet is launched towards the eye using a solenoid. The probe hits the eye and bounces back. The same solenoid, inside which the probe moves, is used to detect the movement and impact of the probe, because the moving magnet induces voltage in the solenoid. In the clinical study, the Goldmann applanation tonometer was used as a standard. Handheld and fixed tonometers were also compared by means of measurements in pressurized pig eyes. RESULTS: At a low intraocular pressure (IOP), there is a longer impact time than at a high IOP. A handheld tonometer causes more variation in impact times, especially at low intraocular pressure (IOP), than a fixed tonometer. CONCLUSIONS: The new induction-based impact method can be used to measure IOP. When a fixed tonometer is used, there is less variation in impact times. PMID- 10794244 TI - Comparison of ranked segment analysis (RSA) and cup to disc ratio in computer assisted optic disc evaluation. AB - PURPOSE: Ranked segment analysis is a new method for evaluation of optic nerve head topography with the Heidelberg Retina Tomograph. This analysis ranks sector measurements around the optic disc and compares these measurements with rank specific significance limits. We evaluated the diagnostic precision of the ranked segment analysis in a large clinical sample and compared with vertical cup/disc ratio measurements. Vertical cup/disc ratio measurements were not corrected for disc size. METHODS: We analysed optic disc images from 153 normal and 75 glaucomatous eyes using the Heidelberg Retina Tomograph (software version 2.01). Ranked segment analyses and vertical cup/disc ratio measurements were obtained from each optic disc image, and ROC curves were plotted. RESULTS: At the 95% specificity level, sensitivity was significantly lower for ranked segment analysis (70%) compared to vertical cup/disc ratio measurements (85%). Almost 30% of the discs in the glaucoma group were classified as normal using the ranked segment analysis. DISCUSSION: Ranked segment analysis yielded unacceptably poor discrimination, in fact, worse than cup/disc ratio with its known clinical limitations Glaucomatous disc damage typically occurs at the vertical poles of the disc. High rank measurements (low sector values) in normal eyes, on the other hand, are more common in the temporal disc sectors. Important spatial information is lost during the ranking procedure. This may partly explain the low sensitivity of the ranked segment analysis observed in our study. The results raise serious concerns regarding the clinical usefulness of ranked segment analysis. PMID- 10794247 TI - NGF administered by microdialysis into rabbit vitreous. AB - PURPOSE: Microdialysis has been used in eye research for almost a decade. We have previously developed and used it mainly for chronic experiments and here it is used in acute experiments. It is highly suitable for both administration and withdrawal of substances from the eye, as it only permits molecules to cross the intraocular membrane without any net fluid movement in or out of the eye. METHODS: We have here investigated the ability of 125I labeled NGF (nerve growth factor) to cross a previously implanted high permeability membrane of a new type (polyether sulphone, PES, cutoff at 100000 daltons) in the rabbit vitreous. It was perfused for different time periods with a solution containing NGF The radioactivity of the entire vitreous was then counted. RESULTS: The results show detectable levels (up to 10(-11)M) of the substance after the perfusions. CONCLUSION: We conclude that microdialysis with PES membranes is a possible method for the intraocular administration of NGF. PMID- 10794246 TI - Identification of androgen, estrogen and progesterone receptor mRNAs in the eye. AB - PURPOSE: Previous research has demonstrated that sex steroids exert a significant influence on the structure and function of numerous ocular tissues. To begin to explore the underlying basis of this hormone action, we examined whether various anterior and posterior tissues of the eye contain androgen, estrogen and progesterone receptor mRNAs. METHODS: Tissue samples were obtained from adult male and female rats, rabbits and humans, processed for the isolation of total RNA and analyzed by RT-PCR, agarose gel electrophoresis and Southern blot hybridization. All PCR amplifications included positive and negative controls. RESULTS: Our findings showed that androgen, estrogen and/or progesterone receptor mRNAs are present in the lacrimal gland, lacrimal gland acinar epithelial cells, meibomian gland, lid, palpebral and bulbar conjunctivae, cornea, iris/ciliary body, lens, retina/uvea, retina/choroid and retinal pigment epithelial cells of rats, rabbits or humans. CONCLUSIONS: Our findings demonstrate that sex steroid receptor mRNAs exist in a variety of ocular tissues and suggest that these sites may represent target organs for androgens, estrogens and/or progestins. PMID- 10794248 TI - Tumors and pseudotumors of the optic disc. AB - There are many lesions that can present as an elevation of the optic disc. Since each of them has different ophthalmic and systemic significance, it is important for the ophthalmologist to recognize them and to plan strategies of management when necessary. Important inflammatory and benign congenital conditions should be differentiated from malignant neoplasms. This review covers the differential diagnosis of lesions that can cause an elevation of the optic disc. It includes selected congenital lesions, inflammatory conditions, non-inflammatory non neoplastic disorders, benign hamartomas, and primary and secondary malignant neoplasms. PMID- 10794249 TI - Colour slides or digital photography in diabetes screening--a comparison. AB - PURPOSE: The aim of this study was to compare digital images with slides in detecting and grading diabetic retinopathy, and to assess the retinopathy screening performed by ophthalmic nurses. METHOD: 283 consecutive patients were examined using digital colour and redfree photography (Topcon Imagenet System 1.53) and 35 mm slides (Topcon TRC-50 VT fundus camera, Kodachrome 64 colour film). The images were graded by the worst eye according to the Wisconsin classification by an ophthalmologist and ophthalmic nurse independently. RESULTS: There was exact agreement between grades obtained from the colour slides and the digital colour images in 82% (weighted kappa 0.88; 95% CI 0.80-0.96), and in 85% when redfree images were used as an adjunct to the digital colour images There was a tendency towards undergrading of the digital colour images and overgrading of the digital redfree images, compared with the colour slides. Inter- and intragrader agreement (weighted kappa) varied between 0.77 and 0.84 for digital photography and between 0.88 and 0.90 for colour slides CONCLUSION: Good to excellent agreement was found between the grading of colour slides and digital colour images, the latter, however, associated with a slightly lower reliability. The adjunct of redfree images seemed to facilitate the detection of retinopathic lesions. PMID- 10794250 TI - Foveal haemorrhages in diabetic retinopathy. Clinical characteristics and visual outcome. AB - BACKGROUND: Haemorrhages from retinal vessels is one of the major clinical characteristics of diabetic retinopathy. Vitreous haemorrhages from retinal neovascularizations may extend to the visual axis and disturb central vision, whereas asymptomatic intraretinal haemorrhages may develop from ruptures of smaller retinal vessels. On rare occasions, however, smaller intraretinal haemorrhages may develop in the fovea, and consequently lead to a reduction in central vision. The clinical characteristics and visual outcome of these lesions have not been described in detail. METHODS: Clinical data of 4724 diabetic patients (31.4% with type 1 diabetes and 68.6% with type 2 or other diabetes types) examined in the screening clinic for diabetic retinopathy at the Department of Ophthalmology, Arhus University Hospital, 1993-1998 were reviewed. Patients who had had a previous foveal haemorrhage were subjected to a full ophthalmological reexamination. RESULTS: Six eyes of six patients with type 1 diabetes had previously had a foveal haemorrhage. The lesion had resulted in a visual reduction of on the average 1.4 visual acuity steps (SD=0.5, range:1-2, n=5), and resolution of the lesion was accompanied by an increase in visual acuity of on the average 1.2 visual acuity steps (SD=0.4, range: 1-2, n=6). Four of the patients had progressed to proliferative diabetic retinopathy and had received pan-retinal photocoagulation. CONCLUSIONS: Foveal haemorrhages in diabetic retinopathy are accompanied by a mild and transient reduction in central vision. The lesions predominate in patients with type 1 diabetes of long duration, and may indicate that retinopathy has developed into a moderate or severe stage. PMID- 10794251 TI - Ophthalmic artery blood flow velocity changes in diabetic patients as a manifestation of macroangiopathy. AB - PURPOSE: The hemodynamic characteristics of ophthalmic artery (OA) blood flow velocity in diabetic patients with ocular involvement were evaluated. METHODS: Changes in OA blood flow of eyes with background diabetic retinopathy (BDR), proliferative retinopathy (PDR) and ocular ischemic syndrome (OIS) were analyzed by Color Doppler imaging. RESULTS: Patients with BDR and PDR had significantly lower diastolic and mean blood flow velocities and higher pulsatility indices compared to controls. Diabetic patients with OIS had significantly lower systolic, diastolic and mean anterograde OA blood flow velocities than the controls. Pulsatility indices were higher in anterograde OA blood flow measurements compared to controls. Systolic blood flow velocities in rubeotic eyes with OIS were significantly lower than in rubeotic eyes with PDR. CONCLUSION: OA blood flow measurements by color doppler imaging may detect macroangiopathies in diabetic patients as manifested by carotid atheromas and arterio- and atherosclerosis of the OA and its branches. PMID- 10794252 TI - Myopisation: the refractive tendency in teenagers. Prevalence of myopia among young teenagers in Sweden. AB - PURPOSE: 1045 children between 12 and 13 years old were examined in a field study in the Goteborg area (Sweden). The aim of this study was to report the prevalence of refractive errors, with special attention to myopia, since there are no previous reports in Sweden about this age group. METHODS: The examination included visual acuity testing and refraction under cycloplegia. RESULTS: We found a prevalence of myopia (> or = -0.5D) of 49.7% and a prevalence of bilateral myopia of 39%. In the whole population, 23.3% were considered to need glasses (> or = -0.75D). We also found a prevalence of high myopia (> or = -5D) in 2.5% of the children. DISCUSSION: No statistically significant difference in myopia with respect to gender was found. The proportion of children needing glasses is considered the best indicator of the prevalence of myopia in this sample. The results confirm that this tendency towards myopisation in a teenage population in Goteborg is similar to the prevalence found in other parts of the world. PMID- 10794253 TI - Retinal detachment after silicone oil removal. AB - PURPOSE: To evaluate the causes of retinal detachment after silicone oil removal, to define possible risk factors and the anatomical and functional prognosis of this complication. METHODS: 112 eyes that underwent silicone oil removal were included. The group of eyes with retinal detachment after oil removal (18/112 eyes, 16.1%) was compared with the group with no postoperative retinal detachment. RESULTS: The most common cause for retinal detachment after oil removal was anterior PVR (77.8%). Initial PVR detachment, advanced PVR stages, anterior PVR, more preceding operations, aphakia/pseudophakia, myopia and shorter duration of the oil tamponade were significantly more represented in the group of eyes with retinal detachment after oil removal. The prognosis of retinal detachment after oil removal is poor. CONCLUSION: Some criteria could be regarded as risk factors for retinal detachment after silicone oil removal. Improvement of the results should be possible by considering these factors and by control of reproliferations. PMID- 10794254 TI - The 30-Hz flicker cone ERG for monitoring the early course of central retinal vein occlusion. AB - PURPOSE: Investigation of the full-field ERG (electoretinography) within 24 hours after the onset of symptoms in central retinal vein occlusion, and repeated examination within the following days and weeks. METHODS: Seven patients with central retinal vein occlusion were examined with full-field ERG within less than 24 hours after the onset of their symptoms and every second to third day during the first 3 weeks. They were then followed for a period of time of six months. RESULTS: The amplitudes as well as the implicit times in the 30 Hz flicker ERG changed considerably within the first three weeks. In the three patients who developed rubeosis, the cone b-wave implicit times as well as the amplitudes were altered considerably. In the patients that did not develop rubeosis, the cone b wave implicit times as well as the amplitudes in the 30 Hz flicker ERG were more stable. CONCLUSION: The full-field cone ERG reflects the retinal function and the ERG parameters change considerably during the first weeks after the onset of the central retinal vein occlusion. The 30 Hz flicker ERG gives important information concerning the early development of the disease. The optimal time to do a predictive ERG could be after three weeks. PMID- 10794255 TI - Surgical effects of combined trabeculotomy and sinusotomy compared to trabeculotomy alone. AB - PURPOSE: To elucidate long-term surgical results and problems of combined trabeculotomy and sinusotomy in comparison with trabeculotomy alone. METHODS: The study included 91 glaucomatous eyes in 57 patients with primary open angle glaucoma, who underwent either combined trabeculotomy and sinusotomy (40 eyes) or trabeculotomy alone (51 eyes). The mean follow-up period (+/-standard deviation) was 46.1+/-10.4 months (range: 30 to 60 months). RESULTS: At the final examination, in 37 (93%) of the 40 eyes after combined trabeculotomy and sinusotomy and in 40 (78%) of the 51 eyes after trabeculotomy alone, intraocular pressures were well controlled. The mean intraocular pressures at the end of the first postoperative year were 15.6+/-3.0 mmHg in the 40 eyes after the combined procedure, and 17.8+/-3.1 mmHg in the 51 eyes after trabeculotomy alone (p=0.0001, unpaired t-test). Complications included Descemet's membrane detachment in 1 eye (3%) and 4 eyes (8%), and transient elevation of intraocular pressure to higher than 30 mmHg (IOP spike) in 6 eyes (15%) and 11 eyes (22%), respectively, after the combined procedure and trabeculotomy alone. CONCLUSION: Combined trabeculotomy and sinusotomy is a useful surgical modality for the treatment of primary open angle glaucoma to obtain lower intraocular pressure levels than those after trabeculotomy alone. PMID- 10794256 TI - Peribulbar anesthesia and sub-Tenon injection for vitreoretinal surgery: 300 cases. AB - BACKGROUND AND OBJECTIVES: We carried out a prospective study in order to evaluate the efficacy and safety of peribulbar anaesthesia supplemented by a sub Tenon injection in case of inadequate analgesia during vitreoretinal surgery. METHODS: We performed 300 consecutive vitreoretinal procedures. Patients received a mean volume of 17+/-4.5 ml of a mixture of etidocaine 1%, bupivacaine 0.50% and hyaluronidase (25 UI/ml). Supplementation was represented by a sub-Tenon infiltration of lidocaine 2% (2 or 3 ml). This volume was not included in the mean volume. RESULTS: Analgesia was adequate throughout surgery without any supplementation in 85% of cases and with a sub-Tenon infiltration in 99%. Akinesia was complete in 82%, mild in 15% and absent in 3% of cases. The sub Tenon injection was performed immediately before starting the procedure in 58% of cases and during the surgery with a delay of 80+/-21 min in 42%. Eleven patients (3.66%) were agitated during surgery and two of them needed a general anaesthesia to allow for the procedure. Generalised epilepsy was encountered in two patients (0.66%) immediately after the peribulbar injection in one patient and 15 min later in the other. The systolic blood pressure severely decreased between 60 to 70 mm Hg 40 min after the accomplishment of the peribulbar in 2 patients and at 90 min in 2 others. CONCLUSION: Our results demonstrate that peribulbar anaesthesia alone offers excellent analgesia in 85% of patients and supplemented by a sub-Tenon injection in 99%. PMID- 10794257 TI - The influence of age, gender, refractive error, and optic disc size on the optic disc configuration in Japanese normal eyes. AB - PURPOSE: To investigate the influence of age, gender, refractive error, and optic disc size on optic disc configuration in Japanese normal eyes. METHODS: Ninety two eyes from 92 visually normal Japanese subjects (mean refractive error+/-SD: 1.26+/-2.25 D, range -8 D to +3 D) were examined using a confocal scanning laser tomograph, TopSS. The following disc parameters were investigated: disc size, total or quadrant C/D area ratio and neuroretinal rim area, half-depth area, volume below, and average cup depth. RESULTS: The disc diameter ( mean+/-SD: 1.84+/-0.16 mm) and disc size showed highly significant correlations with the C/D ratio (p<0.001) and the neuroretinal rim area (p<0.001). No other correlation was observed. CONCLUSION: These results indicate that the optic disc diameter and disc size have higher correlations with the optic disc configuration than age, gender, and refractive error in Japanese. These results are similar to those data reported for eyes of Caucasians or Afro Americans, and should be considered when optic discs are evaluated. PMID- 10794258 TI - Paediatric laser DCR. AB - PURPOSE: Endoscopic laser dacrocystorhinostomy is an alternative procedure to external approach DCR in children. We report our results with 5 patients (6 eyes) with congenital nasolacrimal duct obstruction. METHODS: Six endonasal DCR's were carried out using the KTP laser in patients with congenital nasolacrimal duct obstruction resistant to probing and intubation. RESULTS: The follow-up ranged from 3-8 months. All patients experienced a recurrence of their symptoms postoperatively. CONCLUSION: Endoscopic laser assisted DCR is not a successful procedure in children. PMID- 10794259 TI - Results of transantral orbital decompression in patients with thyroid-associated ophthalmopathy. AB - PURPOSE: To present the results of orbital decompression in patients with thyroid associated ophthalmopathy (TAO). METHODS: Transantral orbital decompression was performed in 63 patients with TAO. In 40 patients (63%) the operation was made because of progressive ophthalmopathy not responding to medical therapy, and in 23 patients (37%) the operation was made for rehabilitative reasons. The long term hypesthesia engaging the infraorbital nerve was assessed with a questionnaire using a Visual Analogue Scale (VAS). RESULTS: The mean proptosis reduction was 3.2 mm (range 0-8 mm). Twenty-one patients had impaired visual acuity preoperatively, and 20 improved. Altogether 30 patients (40%) had worsened ocular motility postoperatively. Forty-three patients did not have diplopia in the primary position preoperatively, and new diplopia developed in 22 of these (51%). Hypesthesia in the infraorbital nerve area was reported for half of the operated sides, but was a major cause of distress (VAS-scoring >5) to eleven patients. CONCLUSIONS: Transantral orbital decompression is indicated in patients with progressive TAO or in patients with prominent exophthalmos, and results in a good proptosis reduction, but the risk of postoperative diplopia is significant. Postoperative hypesthesia is common but often not a major problem. PMID- 10794260 TI - A clinical comparison of the Xpert non-contact tonometer with the Goldmann applanation tonometer after penetrating keratoplasty. AB - PURPOSE: To assess the agreement between the Xpert non-contact tonometer (NCT) and the Goldmann applanation tonometer in patients who have undergone penetrating keratoplasty. METHODS: The study material consisted of 42 consecutive patients (43 eyes) who had undergone penetrating keratoplasty within the previous 13 months. RESULTS: The slope of the linear relationship between the two measurement methods did not differ significantly from 1.0. The mean difference between the methods of 0.96 mmHg was not statistically significant. The range of intraindividual differences between the methods was from -9.8 to 22.8 mmHg. The standard deviation of differences was 6.62 mmHg. The 95% limits of agreement were -12.00 to 13.94 mmHg. There was no significant correlation between the central corneal thickness, astigmatism or transplant size and the difference between the methods CONCLUSION: The Xpert NCT shows considerable variation from the Goldmann values The degree of agreement with the true IOP value remains to be shown. PMID- 10794261 TI - Risk factors for the development of pterygium in Singapore: a hospital-based case control study. AB - PURPOSE: A hospital-based case-control study, designed to evaluate the risk factors for the development of pterygium in Singapore. METHODS: Sixty-one cases and 125 controls were interviewed at the Singapore National Eye Centre. Information on present and past (5 and 10 years ago) exposure to sunlight on weekdays and weekends, sociodemographic data, behavior lifestyle and family history of eye disease was collected. RESULTS: Patients with pterygia were mostly males, smoked and drank alcohol more often, had lower combined family incomes and were more likely to have a family history of eye disease. The adjusted odds ratios for the development of pterygia were 1.27 (95 percent confidence interval 1.06 to 1.54) and 1.31 (95 percent confidence interval 1.09 to 1.57) for participants who had spent more time in the sun five and ten years ago respectively. CONCLUSIONS: Pterygium is an important public health problem in Singapore and its development is related to the amount of sunlight exposure. PMID- 10794262 TI - High myopia, hypertelorism, iris coloboma, exomphalos, absent corpus callosum, and sensorineural deafness: report of a case and further evidence for autosomal recessive inheritance. PMID- 10794263 TI - Malignant changes in a giant orbital keratoacanthoma developing over 25 years. AB - PURPOSE: To report a patient with a history over 25 years of a slowly growing, large, invasive crateriform tumour filling the anterior part of the orbit. METHODS: A 61-year-old male presented with a large tumour of the left orbit. Exenteration was performed with subsequent histological analysis of the excised mass. RESULTS: The main tumour showed the characteristic features of a keratoacanthoma. However, the posterior aspect of the tumour disclosed the morphology of a squamous cell carcinoma. Six months later, the patient presented with metastases to lymph nodes, lung and mediastinal tissue. A leukemoid reaction was diagnosed by fine needle biopsy. CONCLUSION: The giant variety of keratoacanthoma may fail to regress and can transform into a squamous cell carcinoma. In our patient, the development of a chronic lymphoid leukemia raises the possibility that it may be the underlying cause for the transformation of the posterior part of the keratoacanthoma into a frank squamous cell carcinoma. PMID- 10794264 TI - Paediatric chiasmal neuritis--typical of post-Epstein-Barr virus infection? AB - A case of chiasmal neuritis with bilateral, asymmetric optic nerve involvement is presented. Association with recent Epstein-Barr virus infection is suggested. The clinical picture, possible aetiological factors, and causative relation to Epstein-Barr virus are presented. PMID- 10794265 TI - Bilateral necrotizing scleritis and blindness in the myelodysplastic syndrome presumably due to relapsing polychondritis. AB - PURPOSE: The purpose of this study was to report a case of bilateral blindness, bilateral necrotizing scleritis, and bilateral deafness in a patient with myelodysplastic syndrome (MDS). In such a patient, the possibility of relapsing polychondritis (RPC) associated with MDS must be considered. CASE REPORT/METHODS: A 66-year-old patient suffered from myelodysplastic syndrome (MDS). Shortly before his death, he became bilaterally blind and deaf. A biopsy was taken from the conjunctiva and the bone marrow, and both eyes were obtained after death for further investigation. Findings of the clinical and laboratory work-up for the patient's hematologic disorder as well as an examination of the eyes by light microscopy and immunohistochemistry are presented. RESULTS: Ocular sections showed a diffuse necrotizing scleritis with moderate uveitis and no identifiable infectious agent. Neither was there any evidence of a leukemic infiltration. The deafness had been due to inner ear failure, and the patient died of a cardiac failure. CONCLUSIONS: Non-infectious scleritis associated with inner ear deafness is a strong indication of relapsing polychondritis (RPC). Furthermore, RPC can be associated with MDS. Thus, in addition to leukemic infiltration and infection involving ocular structures, ophthalmologists and otolargyngologists should be aware of the association between MDS and RPC and the potential complications. PMID- 10794267 TI - Optic nerve coloboma with retinal degeneration associated with cystic microphthalmia of the other eye. AB - In a seventy-five-year old man an optic nerve coloboma with generalised retinal degeneration associated with a cystic microphthalmia of the other eye is described. The MR imaging revealed the existence of a left microphthalmic eye with a lower lid cyst. From the other eye an optic nerve coloboma with a cystic ectasia of the coloboma area freely open to the vitreous cavity was apparent. The ERG recorded from this eye was extinguished. PMID- 10794266 TI - Central serous chorioretinopathy following pigment epithelium detachment: fluorescein and indocyanine green angiography follow-up. AB - PURPOSE: To evaluate the evolution from idiopathic serous pigment epithelium detachment (PED) to central serous chorioretinopathy (CSC). METHODS: Fluorescein angiography (FA) and indocyanine green angiography (ICGA) were performed using the digital imaging system Topcon IMAGEnet H 1024. RESULTS: A leakage point which later became the typical smokestack image of a CSC was found in the upper margin of the persistent PED. Dilation of the choroidal vessels under the detached neuroepithelium was also seen. CONCLUSIONS: Clinical and angiographic evidence about the relation between the two entities can help us to understand CSC as a potential evolution of PED and to continue searching for the common injury of the pigment epithelium which probably is the primary event. PMID- 10794268 TI - Unexpected intraocular ricochet path of shotgun pellet. PMID- 10794269 TI - A five-dimensional structural investigation of the misfit layer compound AB - The structure of the misfit layer compound [Bi0.87SrO2]2[CoO2]1.82, bismuth strontium cobaltite, was determined by single-crystal X-ray diffraction using the five-dimensional superspace-group formalism. This composite crystal, of monoclinic symmetry, is composed of two subsystems exhibiting incommensurate periodicities along b, the binary axis direction. The first composite part [Bi0.87SrO2] displays an intrinsic modulation of planar monoclinic type characterized by the wavevector q* = 0.293a* + 0.915c*. The second composite part [CoO2] shows two different centered lattice variants. The structure of the misfit layer crystal can be described as an alternation along c of distorted rock-salt type slabs, formed from [BiO] and [SrO] layers (first subsystem), and of [CoO2] layers (second subsystem) displaying a distorted CdI2-type structure. Two main structural results are obtained. First, as a consequence of the intrinsic modulation, disordered zones, characterized by Bi vacancies, are regularly distributed in the [BiO] layers. Second, strong chemical bonds are implied between the strontium atoms of the first subsystem and the oxygen atoms of the second one. PMID- 10794270 TI - Structure of the defect perovskite Ce1/3NbO3: a redetermination by electron and neutron powder diffraction AB - The crystal structure of the defect perovskite Ce1/3NbO3, cerium niobium oxide, has been re-examined by neutron powder and electron diffraction. The results of a powder neutron Rietveld refinement indicate that the structure is monoclinic: space group P2/m with Z = 4, a = 5.5267 (3), b = 7.8824 (2), c = 5.5245 (3) A, beta = 90.294 (1)degrees, V= 240.67 (2) A3 at 298 K with chi2 = 2.570. Previous reports have described the Ce1/3NbO3 structure in a smaller (V/2) orthorhombic cell based solely upon X-ray powder diffraction data. The presence of weak reflections in the electron diffraction pattern provides conclusive evidence for a monoclinic superstructure of the orthorhombic cell. While these superlattice reflections are barely detectable with X-ray radiation, they are clearly visible in the neutron diffraction experiments. The superlattice reflections are shown to arise from a tilting of the NbO6 octahedra which results in the reduction of symmetry from orthorhombic to monoclinic. It is also found that the Ce3+ and Nb5+ cations are displaced from the centres of their respective polyhedra to accommodate the bond-valence requirements of the crystal structure. It is likely that distortions of this type are present in other Ln1/3NbO3 and Ln1/3TaO3 defect perovskites. PMID- 10794271 TI - High-pressure transformations of NbO2F AB - The ReO3-type structure NbO2F, niobium dioxyfluoride, has been studied at high pressures using diamond anvil cells and synchrotron X-ray radiation. High pressure powder diffraction measurements have been performed up to 40.1 GPa. A phase transition from the cubic (Pm3m) ambient pressure structure to a rhombohedral (R3c) structure at 0.47 GPa has been observed. Rietveld refinements at 1.38, 1.96, 3.20, 6.23, 9.00 and 10.5 GPa showed that the transition involves an a-a-a- tilting of the cation-anion coordination octahedra and a change of the anion-anion arrangement to approach hexagonal close packing. Compression and distortion of the Nb(O/F)6 octahedra is also revealed by the Rietveld refinements. At 17-18 GPa, the diffraction pattern disappears and the structure becomes X-ray amorphous. PMID- 10794272 TI - Electron density of KNiF3: analysis of the atomic interactions AB - The topological analysis of the electron density in the perovskite KNiF3, potassium nickel trifluoride, based on the accurate X-ray diffraction data, has been performed. The topological picture of the atomic interactions differs from that resulting from the classic crystal chemistry consideration. The shapes of atoms in KNiF3 defined by zero-flux surfaces in the electron density are, in general, far from spherical. At the same time, their asphericity in the close packed layer is very small. The topological coordination numbers of K and Ni are the same as the geometrical ones, whereas topological coordination for the F atom (6) differs from the geometrical value. The latter results from a specific shape of the Ni-atom basin preventing the bond-path formation between F atoms in the same atomic close-packed layer, in spite of the fact that the closest F-F distance is the same as K-F. Judging by the electron density value and curvature at the bond critical points, the K-F interaction in KNiF3 can be considered ionic, while the Ni-F bond belongs to the polar covalent type. No correlation of the topological ionic radii with crystal or ionic radii was found in KNiF3. Critical points in the electrostatic potential have also been studied. PMID- 10794273 TI - Structure of the alums. I. On the sulfate group disorder in the alpha-alums AB - The crystal structures at room (296 K) and low (173 K) temperature of several alpha-alums have been refined by single-crystal X-ray structure analysis. Many alpha-alums of known structure are disordered, the sulfate anions occupying one of two possible sites. All those studied here exhibited such disorder and the relative occupancies of the two sites are in excellent agreement with those obtained by Raman spectroscopy, where the nu1(SO4) mode is seen as a doublet owing to the presence of two different types of sulfate ion. No phase transitions were noted on cooling but there is less disorder. PMID- 10794274 TI - Two phases of AB - The solvent-bridged caesium phosphide [CsPH(eta6-2,4,6-(t)Bu3C6H2)]2(eta3 toluene)0.5x, catena-[(micro-eta3-toluene)-bis[caesium(2,4,6-tri-tert butylphenylphosphide)]], undergoes a reversible solid-state, order-disorder phase transition characterized by the doubling of the unit-cell volume at low temperature achieved by doubling one unit-cell vector. The unit-cell parameters at 293 (2) K (form A) are: a = 11.147 (4), b = 14.615 (4), c = 14.806 (5) A, alpha = 70.57 (3), beta = 71.85 (3), gamma = 72.93 (2)degrees, V= 2112.5 (12) A3, Z = 2, rho(calc) = 1.362 g cm(-3), R1 = 0.0513 for 5462 reflections, wR2 = 0.0947 for all data. The unit-cell parameters at 173 (2) K (form B) are: a = 14.6241 (3), b = 14.7393 (3), c = 22.0720 (4) A, alpha = 72.2117 (7), beta = 73.3659 (8), gamma = 70.2953 (7)degrees, V = 4174.8 (2) A3, Z = 4, rho(calc) = 1.379 g cm(-3), R1 = 0.0405 for 14,010 reflections, wR2 = 0.1326 for all data. With a minor change, the key structural features discussed previously for form A [Rabe et aL (1998). Inorg. Chem. 37, 4235-4245] remain unchanged. The eta3-toluene ligand is observed to be disordered at 293 (2) K and ordered at 173 (2) K, with the order disorder phase transition occurring at approximately 278 (2) K. PMID- 10794275 TI - X-ray diffraction study of the phase transitions of (CH3)4NCdCl3 between 293 and 80 K: a quantitative analysis of the ferroelastic domains distribution below 118 K AB - X-ray diffraction patterns of [N(CH3)4][CdCl3], tetramethylammonium trichlorocadmate(II), have been investigated in the temperature range 80-293 K, which includes two phase transitions at 118 and 104 K, respectively. The main interest in this compound is to establish the mechanism of the structural phase transitions common to other members of the isostructural family [(CH3)4N][MX3]. It is supposed to be related to the ordering of the organic part together with some small distortion of the inorganic chains. The origin of the order-disorder mechanism would be the orientationally disordered distribution of the tetramethylammonium tetrahedra at room temperature. Maximum Entropy Methods suggest that the most probable distribution of the organic groups can be described through the so-called two-well model, in which one threefold axis of the tetramethylammonium tetrahedron coincides with the crystallographic threefold axis of the structure. Below 118 K the reflections are split. However, the splitting cannot be fully explained by the ferroelastic domains expected to appear after the phase transitions. Recent NMR results [Mulla-Osman et al. (1998). J. Phys. Condensed Matter, 10, 2465-2476] corroborate the existence of more domains than expected from symmetry considerations. A model of ferroelastic domains which is in agreement with both X-ray diffraction diagram and NMR measurements is proposed. PMID- 10794276 TI - Packing effects on the geometry of neutral platinum(II) complexes due to solvate molecules: the structures of trans-dichlorobis(triphenylarsine)-platinum(II) AB - A series of structures of trans-dichlorobis(triphenylarsine)platinum(II), recrystallized from four different solvents, have been characterized by X-ray crystallography and were shown to crystallize as different solvates (same metal complex, different crystallization solvents). Their geometric differences induced by packing and solvent molecules were analysed with half-normal probability plots and root-mean-square deviations. The recrystallization solvents used in the investigation were 1,1,1-trichloroethane, dichloromethane, 1,2-dichloroethane and benzene, and the following crystallization modes were obtained. From 1,1,1 trichloroethane the metal complex crystallizes without solvent as trans [PtCl2(AsPh3)2] in P2(1)/n with Z = 2, a = 9.271 (2), b = 19.726 (4), c = 9.830 (2) A, beta = 111.83 (3)degrees, V = 1668.8 (6) A3, R = 0.0262, and from dichloromethane with two solvent molecules as trans-[PtC12(AsPh3)2].2CH2C12 in Pbca with Z= 4, a = 20.582 (4), b = 8.146 (2), c = 23.491 (5) A, V = 3938.5 (14) A3 and R = 0.0316. From dichloroethane it crystallizes with one solvent molecule as trans-[PtC12(AsPh3)2].C2H4C12 in P1 with Z = 1, a = 9.390 (2), b= 9.548 (2), c = 11.931 (2) A, alpha = 109.70 (3), beta = 108.26 (3), gamma = 98.77 (3) , V= 915.6 (3) A3, R = 0.0390, and from benzene with half a solvent molecule as trans- [PtC12(AsPh3)2].0.5C6H6 in P2(1)/n with Z = 4, a = 11.778 (2), b = 18.712 (4), c = 16.647 (3) A, beta = 104.78 (3) , V= 3547.3 (12) A3 and R = 0.0303. In all four compounds platinum(II) coordinates to triphenylarsine and chloride in a pseudo square-planar trans configuration. The Pt-As distances are in the range 2.4104 (4)-2.3923 (4) A and the Pt-C distances are in the range 2.309 (2)-2.2839 (9) A. The solvents have a large influence on the packing, resulting in different space groups or different occupancies in the same space group. Half-normal probability plots show that the largest geometric differences, within the metal complex, are in the bond and torsion angles around the As-C bonds. Very similar torsion angles were observed around the Pt-As bond for all the structures, except for one AsPh3 ligand in the benzene solvate, which differs by about 10 from the others. The metal-donor bond distance varies by as much as 0.019 and 0.025 A (95% confidence interval) for Pt-As and Pt-C1, respectively. The variations are essentially caused by intermolecular interactions. Packing efficiency is expressed as the volume filled by each metal complex in the unit cell and is calculated by subtracting the sum of the solvent molecule volumes from the total volume of the unit cell and then dividing by Z. The efficiency is largest in the dichloroethane solvate and smallest in the non-solvated compound, with a difference of approximately 22 A3 per metal complex. PMID- 10794277 TI - Dimorphism and inclusion compounds of N,N'-di(benzenesulfonyl)-p-phenylenediamine AB - Crystal structures of a new polymorph of N,N'-(p phenylene)bis(benzenesulfonamide) (C18H16N2O4S2) and of two inclusion compounds with acetone [(CH3)2CO] and dimethyl sulfoxide [2(CH3)2SO], respectively, have been determined at 150 K. For a more reliable comparison, the structure of the already known polymorph of N,N'-di(benzenesulfonyl)-p-phenylenediamine has been redetermined under identical conditions. In addition, the phase transformation behavior has been examined by differential scanning calorimetry and the crystallization conditions of both polymorphs including their formation by weathering of the inclusion compounds were investigated. PMID- 10794278 TI - Direct observation of deuterium migration in crystalline-state reaction by single crystal neutron diffraction. II. 3-1 photoisomerization of a cobal-oxime complex AB - Single crystal neutron diffraction analysis of photo-exposed (3-cyanopropyl d2(alpha,alpha))-[(R)-1-phenylethylamine-d11]bis(dimethylglyoximato d14)cobalt(III) was carried out in order to clarify the mechanism of the crystalline-state photoisomerization of the 3-cyanopropyl group bonded to the Co atom in some cobaloxime complexes. Before irradiation the two H atoms bonded to the C1 atom of the 3-cyanopropyl group were exchanged with the D atoms such as - CH2CH2CD2CN. On exposure to a xenon lamp, the cell dimensions of the crystal were gradually changed. After 7 d exposure the change became insignificantly small. The structure was analyzed by neutron diffraction. The 3-cyanopropyl group was transformed to the 1-cyanopropyl group such as --CD(CN)C(H1/2,D1/2)2CH3 with retention of the single-crystal form. This indicates that one of the D atoms bonded to C1 migrates to either position bonded to C2. The other atoms of the complex remained unchanged. These results indicate that photoisomerization proceeded in two steps: the 3-cyanopropyl group was isomerized to the 2 cyanopropyl group in the first place and then the 2-cyanopropyl group was transformed to the 1-cyanopropyl group. Moreover, it was made clear that the second-step isomerization was irreversible, since one of the D atoms was retained. The disordered structure at C2 is estimated to be caused by the interconversion between the 1-cyanopropyl group produced and its dehydrogenated olefin after the photoisomerization. PMID- 10794279 TI - Short N+--H...Ph hydrogen bonds in ammonium tetraphenylborate characterized by neutron diffraction AB - The crystal structures of ammonium tetraphenylborate, NH4+.BPh4-, are determined by neutron diffraction at 20 and 293 K. At both temperatures, all four N-H vectors of the ammonium ion are time-average-oriented at the midpoints of the phenyl rings of neighboring anions. The N-H...Ph distances, H...M 2.067 and N...M 3.023 A, are exceptionally short (M = aromatic midpoint). Even at 20 K the ammonium ion performs large amplitude motions which allow the N-H vectors to sample the entire face of the aromatic system. PMID- 10794280 TI - Hydrogen-bonding behaviour of benzene-1,2,4,5-tetracarboxylic acid: supramolecular structures of different dimensionality in the 2:1 adducts formed with 4,4'-bipyridyl and hexamethylenetetramine AB - Co-crystallization of benzene-1,2,4,5-tetracarboxylic acid, C10H6O8, with 4,4' bipyridyl, C10H8N2, or with hexamethylenetetramine, C6H12N4, from methanol solutions yields in each case a 2:1 salt, [(C10H9N2)+]2.[(C10H4O8)2-] (1) and [(C6H13N4)+]2.[(C10H4O8)2-] (2). In (1) the carboxylate anions lie across centres of inversion, but they contain no intramolecular O-H...O hydrogen bonds: the cations and anions are linked by strong O-H...N and N-H...O hydrogen bonds into a chain-of-rings, and these chains are further linked into a three-dimensional framework structure by means of C-H...O hydrogen bonds and aromatic pi...pi stacking interactions. Compound (2) contains two independent three-molecule aggregates, comprising a central anion and two cations, linked to the anion by means of short N-H...O hydrogen bonds. One of these aggregates is centrosymmetric, but the other is not, and the two types of anion both form two intramolecular O-H...O hydrogen bonds. The two types of three-molecule aggregate, in which all the anions are virtually parallel, are linked by short C-H...O hydrogen bonds into a molecular staircase. PMID- 10794281 TI - Oligosiloxanediols as building blocks for supramolecular chemistry: hydrogen bonded adducts with amines form supramolecular structures in zero, one and two dimensions AB - The structure of 1,1,3,3,5,5-hexaphenyltrisiloxane-1,5-diol-pyrazine (4/1), (C36H32O4Si3)4.C4H4N2 (1), contains finite centrosymmetric aggregates; the diol units form dimers, by means of O-H...O hydrogen bonds, and pairs of such dimers are linked to the pyrazine by means of O-H...N hydrogen bonds. In 1,1,3,3,5,5 hexaphenyltrisiloxane-1,5-diol-pyridine (2/3), (C36H32O4Si3)2.(C5H5N)3 (2), the diol units are linked into centrosymmetric pairs by means of disordered O-H...O hydrogen bonds: two of the three pyridine molecules are linked to the diol dimer by means of ordered O-H...N hydrogen bonds, while the third pyridine unit, which is disordered across a centre of inversion, links the diol dimers into a C3(3) (9) chain by means of O-H...N and C-H...O hydrogen bonds. In 1,1,3,3 tetraphenyldisiloxane-1,3-diol-hexamethylenetetramine (1/1), (C24H22O3Si2).C6H12N4 (3), the diol acts as a double donor and the hexamethylenetetramine acts as a double acceptor in ordered O-H...N hydrogen bonds and the structure consists of C2(2) (10) chains of alternating diol and amine units. In 1,1,3,3-tetraphenyldisiloxane-1,3-diol-2,2'-bipyridyl (1/1), C24H22O3Si2.C10H8N2 (4), there are two independent diol molecules, both lying across centres of inversion and therefore both containing linear Si-O-Si groups: each diol acts as a double donor of hydrogen bonds and the unique 2,2'-bipyridyl molecule acts as a double acceptor, thus forming C2(2) (11) chains of alternating diol and amine units. The structural motif in 1,1,3,3-tetraphenyldisiloxane-1,3 diol-pyrazine (2/1), (C24H22O3Si2)2. C4H4N2 (5), is a chain-of-rings: pairs of diol molecules are linked by O-H...O hydrogen bonds into centrosymmetric R2(2) (12) dimers and these dimers are linked into C2(2) (13) chains by means of O H...N hydrogen bonds to the pyrazine units. 1,1,3,3-Tetraphenyldisiloxane-1,3 diol-pyridine (1/1), C24H22O3Si2.C5H5N (6), and 1,1,3,3-tetraphenyldisiloxane-1,3 diol-pyrimidine (1/1), C24H22O3Si2.C4H4N2 (7), are isomorphous: in each compound the amine unit is disordered across a centre of inversion. The diol molecules form C(6) chains, by means of disordered O-H...O hydrogen bonds, and these chains are linked into two-dimensional nets built from R6(6) (26) rings, by a combination of O-H...N and C-H...O hydrogen bonds. PMID- 10794282 TI - 3,6,9,16,19,22-Hexaazatricyclo AB - The adduct (1) formed between the hexaaza macrocycle 3,6,9,16,19,22 hexaazatricyclo[22.2.2.2(11,14]triconta-1(26),11(29),12,14(30),24,27-hexaene, C24H38N6, and 4,4'-sulfonyldiphenol, O2S(C6H4OH)2, is a salt [(C24H40N6)2+].2[(HOC6H4SO2C6H4O)-], and the adduct (2) formed by the same macrocyclic amine with 4,4'-biphenol is an aquated salt which also contains neutral biphenol molecules, [(C24H40N6)2+]. 2[(HOC6H4C6H4O) ].(HOC6H4C6H4OH).2H2O. In both compounds the cations lie across centres of inversion: there are two crystallographically distinct cation sites in (1) and the conformations of the cations occupying them are quite different. In (2) the single type of cation site is occupied by a conformationally disordered cation: the major and minor components represent two further distinct conformers. In (1) the anions are linked by O-H...O hydrogen bonds into chains, and each cation is linked by a total of six N-H...O hydrogen bonds to anions in four different chains, so linking the chains into continuous sheets. In (2) the anions and the water molecules are linked into sheets, which are further linked into a continuous three-dimensional framework by both the cations and the neutral biphenol units. PMID- 10794284 TI - Bulk modulus and high-pressure crystal structures of tetrakis(trimethylsilyl)methane C AB - The pressure dependence of the crystal structure of cubic tetrakis(trimethylsilyl)methane C[Si(CH3)3]4 (TC) (P < 16.0 GPa, T = 298 K) is reported using high-resolution angle-dispersive X-ray powder diffraction. The compound has crystal structures with the molecules in a cubic-close-packed (c.c.p.) arrangement. It shows three phase transitions in the measured pressure range. At ambient conditions, TC has space group Fm3m (Z = 4) with a = 12.8902 (2) A, V = 2141.8 (1) A3 (phase I). Between 0 and 0.13 GPa TC exhibits a first order phase transition into a structure with space group Pa3 (phase II). A second first-order phase transition occurs between 0.2 and 0.28 GPa into a structure with space group P2(1)3 (phase III). Under non-hydrostatic pressure conditions (P > 10 GPa) a transformation is observed into a c.c.p. structure that is different from the face-centred-cubic structure at ambient conditions. A non-linear compression behaviour is observed, which could be described by a Vinet relation in the range 0.28-4.8 GPa. The extrapolated bulk modulus of the high-pressure phase III was determined to be K0 = 7.1 (8) GPa. The crystal structures in phase III are refined against X-ray powder data measured at several pressures between 0.49 and 4.8 GPa, and the molecules are found to be fully ordered. This is interpreted to result from steric interactions between neighbouring molecules, as shown by analysing the pressure dependence of intramolecular angles, torsion angles and intermolecular distances. Except for their cell dimensions, phases I, II and III are found to be isostructural to the corresponding phases at low temperatures. PMID- 10794283 TI - Anisotropic crystal structure distortion of the monoclinic polymorph of acetaminophen at high hydrostatic pressures. AB - The anisotropy of structural distortion of the monoclinic polymorph of acetaminophen induced by hydrostatic pressure up to 4.0 GPa was studied by single crystal X-ray diffraction in a Merrill-Bassett diamond anvil cell (DAC). The space group (P2(1)/n) and the general structural pattern remained unchanged with pressure. Despite the overall decrease in the molar volume with pressure, the structure expanded in particular crystallographic directions. One of the linear cell parameters (c) passed through a minimum as the pressure increased. The intramolecular bond lengths changed only slightly with pressure, but the changes in the dihedral and torsion angles were very large. The compressibility of the intermolecular hydrogen bonds NH...O and OH...O was measured. NH...O bonds were shown to be slightly more compressible than OH...O bonds. The anisotropy of structural distortion was analysed in detail in relation to the pressure-induced changes in the molecular conformations, to the compression of the hydrogen-bond network, and to the changes in the orientation of molecules with respect to each other in the pleated sheets in the structure. Dirichlet domains were calculated in order to analyse the relative shifts of the centroids of the hydrogen-bonded cycles and of the centroids of the benzene rings with pressure. PMID- 10794285 TI - Molecular analysis of the beta-polymorphic form of trielaidin: crystal structure at low temperature AB - This work reports on the structure of trielaidin [EEE, 1,2,3-tri(trans-9 octadecenoyl)glycerol], a trans unsaturated triglyceride present in many refined fatty materials (margarines, chocolate products etc.). Firstly, the polymorphism, i.e. the existence of different crystalline forms at various temperature ranges, was defined. Secondly, the crystal growth was examined. By developing a particular growing system, monocrystals of the most stable polymorphic form, i.e. the beta-form, were obtained. To reduce thermal vibrations the X-ray data were collected at low temperature (173 K) and the structure was solved using direct methods. The structure was then analyzed in terms of conformation and crystal packing and compared with those of the other known triglycerides. PMID- 10794286 TI - Structure of the inclusion complex of beta-cyclodextrin with 1,12-dodecanedioic acid using synchrotron radiation data; a detailed dimeric beta-cyclodextrin structure AB - A detailed crystal structure study of the dimeric inclusion complex of beta cyclodextrin (betaCD) with 1,12-dodecanedioic acid is presented [IUPAC name: beta cyclodextrin-1,12-dodecanedioic acid (2/1)]. The structure was solved with synchrotron high-resolution data (0.65 A) at 100 K [crystal data: P1, Z= 1, a = 18.153 (7), b = 15.456 (8), c = 15.251 (4) A, alpha = 102.81 (2), beta = 113.13 (2), gamma = 99.90 (3)degrees, V = 3,673 (3) A3, R = 0.0474 for 25,134 unique reflections with I > 2sigma(I)]. Moreover, the room-temperature structure is used for comparison [crystal data: P1, Z = 1, a = 18.220 (3), b = 15.488 (3), c = 15.409 (3) A, alpha = 102.903 (6), beta = 113.122 (5), gamma = 99.708 (5)degrees, V = 3735.2 (12) A3, R = 0.0828 for 8,235 unique reflections with I > 2sigma(I)]. Combining the high-resolution data and the low-temperature made possible the location of the disordered guest molecule, 1,12-dodecanedioic acid, inside the wide cavity of the macrocycle formed by two betaCD monomers. Moreover, almost all the H atoms of the betaCD macrocycle and many of the water molecules have been located in the low-temperature structure. Thus, for the first time, it has been possible to show in detail, up to now only given by neutron diffraction data, that two betaCD monomers self-assemble through O3...O3 intermolecular hydrogen bonds to form the betaCD dimer, as well as describe the hydrogen-bonding scheme between the dimer's hydroxyl groups among themselves and with water molecules in the lattice. The long guest threads through two host molecules forming a [3]pseudorotaxane. Its polar carboxyl groups, fully hydrated at the primary faces of the betaCD dimers, influence their packing so that those faces are exposed to the solvent. This is in contrast to the packing of the beta-cyclodextrin complexes of the corresponding aliphatic monoacids, where the dimeric complexes form channels in order to isolate the terminal methyl group from the water environment of the lattice. PMID- 10794287 TI - Gamma surgery for vestibular schwannoma. AB - OBJECT: The goal of this study was to assess the results of gamma surgery (GS) for vestibular schwannoma (VS) in 200 cases treated over the last 10 years and to review the role of this neurosurgical procedure in the management of VS. METHODS: Follow-up reviews ranging from 1 to 10 years were available in 153 of these patients. Follow-up images in these cases were analyzed using computer software that we developed to obtain volume measurements for the tumors, and the clinical condition of the patients was assessed using questionnaires. Gamma surgery was the primary treatment modality in 96 cases and followed microsurgery in 57 cases. Tumors ranged in volume from 0.02 to 18.3 cm(3). In the group in which GS was the primary treatment, a decrease in volume was observed in 78 cases (81%), no change in 12 (12%), and an increase in volume in six cases (6%). The decrease was more than 75% in seven cases. In the group treated following microsurgery, a decrease in volume was observed in 37 cases (65%), no change in 14 (25%), and an increase in volume in six (11%). The decrease was more than 75% in eight cases. Five patients experienced trigeminal dysfunction; in three cases this was transient and in the other two it was persistent, although there has been improvement. Three patients had facial paresis (in one case this was transient, lasting 6 weeks; in one case there was 80% recovery at 18 months posttreatment; and in one case surgery was performed after the onset of facial paresis for presumed increase in tumor size). Over a 6-year period, hearing deteriorated in 60% of the patients. Three patients showed an improvement in hearing. No hearing deterioration was observed during the first 2 years of follow-up review. CONCLUSIONS: Gamma surgery should be used to treat postoperative residual tumors as well as tumors in patients with medical conditions that preclude surgery. Microsurgery should be performed whenever a surgeon is confident of extirpating the tumor with a risk-benefit ratio superior to that presented in this study. PMID- 10794288 TI - Gamma surgery for intracranial metastases from renal cell carcinoma. AB - OBJECT: The goal of this study was to evaluate the effectiveness and limitations of gamma surgery (GS) in the treatment of renal cell carcinoma that has metastasized to the brain. METHODS: The authors performed a retrospective analysis of a consecutive series of 21 patients with 37 metastatic brain deposits from renal cell carcinoma who were treated with GS at the University of Virginia from 1990 to 1999. Clinical data were available in all patients. No patient died of progression of intracranial disease or deteriorated neurologically following GS. Eight patients clinically improved. Follow-up imaging studies were available for 23 tumors in 12 patients. Nine patients did not undergo follow-up imaging. One patient lived 17 months and succumbed to systemic disease: no brain imaging was performed in this case. Another patient refused further imaging and lived 7 months. Seven patients lived up to 4 months after the procedure; however, their physicians did not require these patients to undergo follow-up imaging examinations because of their general conditions-all had systemic progression of disease. Of the 23 tumors that were observed posttreatment, one remained unchanged in volume, 16 decreased in volume, and six disappeared. No tumor progressed at any time, and there were no radiation-induced changes on follow-up imaging an average of 21 months after GS (range 3-63 months). CONCLUSIONS: Gamma surgery provides an alternative to surgical resection of metastatic brain deposits from renal cell carcinoma. Neurological side effects were seen in only one case; freedom from progression of disease was achieved in all cases. PMID- 10794289 TI - Incidence and clinical features of asymptomatic meningiomas. AB - OBJECT: The increased use of computerized tomography (CT) and magnetic resonance (MR) technology has led to an increase in the detection of asymptomatic meningiomas, although the surgical indication for these tumors remains undetermined. The authors investigated the incidence of asymptomatic meningiomas and their clinical features. METHODS: An epidemiological survey was conducted of primary intracranial tumors diagnosed in Kumamoto Prefecture between 1989 and 1996. Follow-up neuroradiological imaging and clinical studies for asymptomatic meningiomas were performed. Primary intracranial tumors were diagnosed in 1,563 residents. Of these lesions, 504 (32.2%) were meningiomas, and of these meningiomas 196 (38.9%) were asymptomatic. The incidence of asymptomatic meningiomas was significantly higher in individuals older than 70 years of age. Furthermore, the incidence of asymptomatic meningiomas was significantly higher in female than in male patients. Of the asymptomatic meningiomas in 196 patients, 87 (44.4%) were surgically removed, whereas 109 (55.6%) were treated conservatively. Of these conservatively treated patients, 63 received follow-up care for more than 1 year. In 20 of these 63 cases, the tumors increased in size over the 27.8-month average follow-up period (range 12-87 months), whereas in the other 43 cases, the tumor size did not increase during a 36.6-month average follow-up period (range 12-96 months). There was no significant difference with respect to age, tumor size, and male/female ratio between the patient group in which the tumor size increased and the group in which it did not increase during the follow-up period. Asymptomatic meningiomas that evidenced calcification on CT scans and/or hypointensity on T2-weighted MR images appear to have a slower growth rate. CONCLUSIONS: Among patients older than age 70 years who underwent operation for asymptomatic meningioma, the neurological morbidity rate was 23.3%; it was 3.5% among younger patients. This indicates that the advisability of surgery in elderly patients with asymptomatic meningiomas must be considered very carefully. PMID- 10794290 TI - Stent-assisted angioplasty of intracranial vertebrobasilar atherosclerosis: an initial experience. AB - OBJECT: Patients with intracranial vertebrobasilar artery (VBA) atherosclerotic occlusive disease have few therapeutic options. Unfortunately, VBA transient ischemic attacks (TIAs) herald a lethal or devastating event within 5 years in 25 to 30% of patients. The authors report their initial experience with eight patients in whom medically refractory TIAs secondary to intracranial posterior circulation atherosclerotic occlusive lesions were treated with stent-assisted angioplasty. METHODS: Eight patients (six men), ranging in age from 43 to 77 years, experienced signs and symptoms of VBA insufficiency despite combination therapy with warfarin and antiplatelet agents. Angiographic studies revealed severe distal vertebral (four patients), proximal basilar (one patient), or proximal and midbasilar stenoses (three patients). Aspirin and clopidogrel were administered for 3 days before primary angioplasty and stent placement, and this regimen was maintained by the patients on discharge. Patients underwent heparinization during the procedure and were given a bolus and 12-hour infusion of abciximab. A neurologist specializing in stroke evaluated all patients before and after the procedure. The VBAs in all patients were successfully revascularized with 7 to 28% residual stenosis. Six patients experienced no neurological complications. One patient died the evening of the procedure due to a massive subarachnoid hemorrhage. Two patients had groin hematomas, one developed congestive heart failure, and one had transient encephalopathy. All surviving patients are asymptomatic up to 8 months postoperatively. CONCLUSIONS: Although primary intracranial VBA angioplasty with stent insertion is technically feasible, complications associated with the procedure can be life threatening. As experience is gained with this procedure, it may be offered routinely as an alternative therapy to patients with medically refractory posterior circulation occlusive disease that may develop into catastrophic VBA insufficiency. PMID- 10794291 TI - Bleeding risk of cerebrovascular malformations in hereditary hemorrhagic telangiectasia. AB - OBJECT: Hereditary hemorrhagic telangiectasia (HHT) is a rare autosomal-dominant vascular dysplasia with a high prevalence of cerebrovascular malformations (CVMs), mostly manifested as arteriovenous malformations (AVMs). The natural history and bleeding risk of these CVMs is unknown. The authors investigated the risk of bleeding in conjunction with clinical and radiological features in patients with HHT and proven CVMs. METHODS: Intravenous digital subtraction (DS) angiography was used to screen 196 patients with HHT for the presence of CVMs. Patients with abnormal results on DS angiography were asked to undergo a conventional cerebral angiographic study. All patients with a proven CVM were assessed by a neurologist. The bleeding risk was retrospectively and prospectively calculated for patients with AVMs only, as well as for the whole cohort of patients with CVMs. Twenty-four patients (12.2%; 16 female and eight male), aged 14 to 66 years (mean 35.4 years) with one or more CVMs were identified. Fifteen patients (62.5%) had a CVM and a pulmonary AVM. Eleven patients (45.8%) exhibited no neurological signs of their CVM; six (25%) had headache or migraine; four (16.7%) had seizures; and three (12.5%) had an intracranial hemorrhage. Twenty-two patients had at least one AVM (with a total of 28 AVMs), whereas two patients only had telangiectases. Twenty-seven AVMs were small (96%), 36% were located in eloquent areas of the brain, and 82% had superficial venous drainage. One third of the patients had multiple CVMs. The bleeding risk for patients with at least one AVM ranged from 0.41 to 0.72% per year, and for the whole cohort the range was 0.38 to 0.69% per year. Calculation of the bleeding risk as determined by lesion-years ranged from 0.36 to 0.56% per year for patients with AVMs and from 0.27 to 0.46% per year for all patients with CVMs. CONCLUSIONS: Patients with HHT have a high risk of harboring a CVM, especially in the presence of a pulmonary AVM. These CVMs are mostly low-grade AVMs (Spetzler-Martin Grade I or II), are frequently multiple, and have a lower risk of bleeding than that associated with sporadic AVMs. Female patients are more often affected than male patients. The inherent low sensitivity of DS angiography screening for CVMs may yield false negative results. PMID- 10794292 TI - Relationships between the epileptic focus and hand area in central epilepsy: combining dipole models and anatomical landmarks. AB - OBJECT: When considering resection of epileptic generators near the central sulcus, it is essential to define the spatial relationship between the epileptic generator and the primary sensorimotor hand area. In this study, the authors assessed the accuracy of dipole modeling of electroencephalographic spikes and median nerve somatosensory evoked potentials (SSEPs) in defining this relationship preoperatively and noninvasively. METHODS: Epileptic spikes and SSEPs in patients with focal central area epilepsy were represented by dipole models coregistered onto global magnetic resonance images. In patients who underwent surgery, spike dipoles were also compared with findings of electrocorticography (ECoG) and with the resection area. To improve the accuracy of the dipole models, anatomical landmarks of the hand area were used to assess the error in SSEP dipole location, and this error measure was used to correct the location of spike dipoles. Five patients with central epilepsy were studied, three of whom underwent ECoG-guided surgical resections. The location of SSEP dipoles correlated well with anatomical landmarks of the primary sensory hand area. The relative position of the spike and SSEP dipoles correlated well with the patients' ictal symptoms, ECoG findings, and the location of the epileptic focus (as defined by the resection cavity in patients who became seizure free postoperatively). Corrected spike dipoles were located even closer to the resection cavity. CONCLUSIONS: The calculation of the relative location of spike and SSEP dipoles is a simple noninvasive method of determining the relationship between the primary hand area and an epileptic focus in the central area. The spatial resolution of this technique can be further improved using easily identifiable anatomical landmarks. PMID- 10794293 TI - Evaluation of a method for noninvasive intracranial pressure assessment during infusion studies in patients with hydrocephalus. AB - OBJECT: A mathematical model previously introduced by the authors allowed noninvasive intracranial pressure (nICP) assessment. In the present study the authors investigated this model as an aid in predicting the time course of raised ICP during infusion tests in patients with hydrocephalus and its suitability for estimating the resistance to outflow of cerebrospinal fluid (Rcsf). METHODS: Twenty-one patients with hydrocephalus were studied. The nICP was calculated from the arterial blood pressure (ABP) waveform by using a linear signal transformation, which was dynamically modified by the relationship between ABP and cerebral blood flow velocity. This model was verified by comparison of nICP with "real" ICP measured during lumbar infusion tests. In all simulations, parallel increases in real ICP and nICP were evident. The simulated Rcsf was computed using nICP and then compared with Rcsf computed from real ICP. The mean absolute error between real and simulated Rcsf was 4.1 +/- 2.2 mm Hg minute/ml. By the construction of simulations specific to different subtypes of hydrocephalus arising from various causes, the mean error decreased to 2.7 +/- 1.7 mm Hg minute/ml, whereas the correlation between real and simulated Rcsf increased from R = 0.73 to R = 0.89 (p < 0.001). CONCLUSIONS: The validity of the mathematical model was confirmed in this study. The creation of type-specific simulations resulted in substantial improvements in the accuracy of ICP assessment. Improvement strategies could be important because of a potential clinical benefit from this method. PMID- 10794294 TI - Efficacy of the Ghajar Guide revisited: a prospective study. AB - OBJECT: The goal of this study was to compare the freehand technique of catheter placement using external landmarks with the technique of using the Ghajar Guide for this procedure. The placement of a ventricular catheter can be a lifesaving procedure, and it is commonly performed by all neurosurgeons. Various methods have been described to cannulate the ventricular system, including the modified Friedman tunnel technique in which a soft polymeric tube is inserted through a burr hole. Paramore, et al., have noted that two thirds of noninfectious complications have been related to incorrect positioning of the catheter. METHODS: Forty-nine consecutive patients were randomized between either freehand or Ghajar Guide-assisted catheter placement. The target was the foramen of Monro, and the course was through the anterior horn of the lateral ventricle approximately 10 cm above the nasion, 3 cm from the midline, to a depth of 5.5 cm from the inner table of the skull. In all cases, the number of passes was recorded for successful cannulation, and pre- and postplacement computerized tomography scans were obtained. Calculations were performed to determine the bicaudate index and the distance from the catheter tip to the target point. CONCLUSIONS: Successful cannulation was achieved using either technique; however, the catheters placed using the Ghajar Guide were closer to the target. PMID- 10794295 TI - Thymidine kinase activation of ganciclovir in recurrent malignant gliomas: a gene marking and neuropathological study. AB - OBJECT: The gene therapy paradigm of intratumoral activation of ganciclovir (GCV) following transduction of tumor cells by retroviral vectors bearing the thymidine kinase (tk) gene has produced dramatic remissions of malignant gliomas in animal models. In human trials, although the technique has been deemed safe, little antitumor effect has been demonstrated. To evaluate the basis of this inefficacy in human gliomas, the authors conducted a gene-marking trial involving neuropathological and biochemical studies of treated tumor specimens. METHODS: Five patients with malignant recurrent gliomas underwent stereotactic biopsy sampling and intratumoral implantation procedures with three aliquots of 10(6) vector-producing cells (VPCs) in columns. After 5 days, the tumor was resected and the tumor bed reimplanted with VPCs, and a course of GCV was given. Patients received clinical and radiological follow up for 6 months. Tumor specimens were analyzed neuropathologically and for tk gene expression by anti-TK immunohistochemistry and TK enzymatic activity. Four patients tolerated the treatment well but experienced tumor progression. The other developed an abscess after the second operation and died. Increased TK enzymatic activity was demonstrated in the one tumor specimen analyzed. Immunohistochemical evidence of tk gene expression was limited to VPCs. Transduction of tumor cells was not seen. Viable tumor cells were seen near VPCs containing TK. The lymphocytic immune response was mild. CONCLUSIONS: Except for the risk of infection inherent in reoperation, this tk-GCV paradigm was both feasible and safe. Pathological studies indicated that limited dissemination of VPCs and vector from the infusion site and failure to transduce tumor cells with the tk gene are major barriers to efficacy. PMID- 10794296 TI - Telovelar approach to the fourth ventricle: microsurgical anatomy. AB - OBJECT: In the past, access to the fourth ventricle was obtained by splitting the vermis or removing part of the cerebellum. The purpose of this study was to examine the access to the fourth ventricle achieved by opening the tela choroidea and inferior medullary velum, the two thin sheets of tissue that form the lower half of the roof of the fourth ventricle, without incising or removing part of the cerebellum. METHODS: Fifty formalin-fixed specimens, in which the arteries were perfused with red silicone and the veins with blue silicone, provided the material for this study. The dissections were performed in a stepwise manner to simulate the exposure that can be obtained by retracting the cerebellar tonsils and opening the tela choroidea and inferior medullary velum. CONCLUSIONS: Gently displacing the tonsils laterally exposes both the tela choroidea and the inferior medullary velum. Opening the tela provides access to the floor and body of the ventricle from the aqueduct to the obex. The additional opening of the velum provides access to the superior half of the roof of the ventricle, the fastigium, and the superolateral recess. Elevating the tonsillar surface away from the posterolateral medulla exposes the tela, which covers the lateral recess, and opening this tela exposes the structure forming the walls of the lateral recess. PMID- 10794297 TI - Quantitative description of the far-lateral transcondylar transtubercular approach to the foramen magnum and clivus. AB - OBJECT: The purpose of this study was to evaluate the far-lateral transcondylar transtubercular approach (complete FLA) based on quantitative measurements of the exposure of the foramen magnum and petroclival area obtained after each successive step of this approach. METHODS: The complete FLA was reproduced in eight specially prepared cadaveric heads (a total of 15 sides). The approach was divided into six steps: 1) C-1 hemilaminectomy and suboccipital craniectomy with unroofing of the sigmoid sinus (basic FLA); 2) partial resection of the occipital condyle (up to the hypoglossal canal); 3) removal of the jugular tuberculum; 4) mastoidectomy (limited to the labyrinth and the fallopian canal) and retraction of the sigmoid sinus; 5) resection of the lateral mass of C-1 with mobilization of the vertebral artery; and 6) resection of the remaining portion of the occipital condyle. After each successive step, a standard set of measurements was obtained using a frameless stereotactic device. The measurements were used to estimate two parameters: the size of the exposed petroclival area and the size of a spatial cone directed toward the anterior rim of the foramen magnum, which depicts the amount of surgical freedom available for manipulation of instruments. The initial basic FLA provided exposure of only 21 +/- 6% of the petroclival area that was exposed with the full, six-step maximally aggressive (complete) FLA. Likewise, only 18 +/- 9% of the final surgical freedom was obtained after the basic FLA was performed. Each subsequent step of the approach increased both petroclival exposure and surgical freedom. The most dramatic increase in petroclival exposure was noted after removal of the jugular tuberculum (71 +/- 12% of final exposure), whereas the least improvement in exposure occurred after the final step, which consisted of total condyle resection. CONCLUSIONS: The complete FLA provides wide and sufficient exposure of the foramen magnum and lower to middle clivus. The complete FLA consists of several steps, each of which contributes to increasing petroclival exposure and surgical freedom. However, the FLA may be limited to the less aggressive steps, while still achieving significant exposure and surgical freedom. The choice of complete or basic FLA thus depends on the underlying pathological condition and the degree of exposure required for effective surgical treatment. PMID- 10794298 TI - Telomerase activity and expression of the telomerase catalytic subunit, hTERT, in meningioma progression. AB - OBJECT: In recent reports, 6 to 19% of meningiomas have been classified as atypical or anaplastic/malignant. Some atypical and anaplastic meningiomas appear to arise from benign tumors by progression. Telomerase activation has recently been associated with malignant progression of human tumors. The authors have investigated a series of benign, atypical, and anaplastic/malignant meningiomas for telomerase activity and expression of the telomerase catalytic subunit human telomerase reverse transcriptase (hTERT). METHODS: A quantitative telomeric repeat amplification protocol was used to detect telomerase enzyme activity in seven (21%) of 34 benign, but in nine (75%) of 12 atypical and in seven (100%) of seven anaplastic/malignant meningiomas. Very high levels of telomerase activity were observed only in highly aggressive tumors. Messenger (m)RNA expression of the catalytic subunit hTERT was found in 11 (33%) of 33 benign, 12 (92%) of 13 atypical, and all seven anaplastic/malignant tumors. All telomerase-positive lesions were also positive for hTERT mRNA, whereas no telomerase activity was detected in six (21%) of 29 hTERT-positive tumors. This indicates that upregulation of hTERT is the rate-limiting step for telomerase activation in the majority of meningiomas. Expression of telomerase and hTERT was seen in all four tumors with gross brain invasion. All recurrent tumors or meningiomas recurring during follow up expressed hTERT. CONCLUSIONS: The results are consistent with a role for telomerase activation during the development of malignancy in meningiomas. Hence, expression of telomerase activity and hTERT might prove to be potentially useful markers for the evaluation of these tumors. PMID- 10794300 TI - Neuroprotective effect of postischemic administration of progesterone in spontaneously hypertensive rats with focal cerebral ischemia. AB - OBJECT: Exogenous progesterone has been shown to reduce brain edema and ischemia induced cell damage and to improve physiological and neurological function during the early stage of focal cerebral ischemia. In the present study, the authors assessed the neuroprotective potential of progesterone during the late stage of ischemia in a transient middle cerebral artery (MCA) occlusion model in the rat. METHODS: Forty-eight male spontaneously hypertensive rats were randomly assigned to six groups. Progesterone was dissolved in dimethyl sulfoxide (DMSO). In four groups of rats, the dissolved progesterone (4 mg/kg or 8 mg/kg) was administered for 2 or 7 days after ischemia. In two control groups DMSO was administered for 2 or 7 days after ischemia. Occlusion of the MCA was induced by insertion of an intraluminal suture, and reperfusion was accomplished by withdrawal of the suture. Treatment was initiated on reperfusion, which followed 2 hours of MCA occlusion, and continued once a day. Lesion volume, neurological deficit, and body weight loss were measured 2 or 7 days after ischemia, depending on the animal group. Treatment with a high dose of progesterone (8 mg/kg) resulted in reductions in lesion size, neurological deficits, and body weight, compared with control rats. CONCLUSIONS: Administration of progesterone to male rats 2 hours after MCA occlusion reduces ischemic brain damage and improves neurological deficit even 7 days after ischemia. PMID- 10794299 TI - Usefulness of postischemic thrombolysis with or without neuroprotection in a focal embolic model of cerebral ischemia. AB - OBJECT: Recent studies have shown that the use of thrombolysis in the setting of acute stroke is associated with an increased risk of cerebral hemorrhage. The time of onset of symptoms to initiation of medication and the dose levels of the thrombolytic agents are important determinants for the risk of cerebral hemorrhage. The authors evaluated the time course of thrombolysis-related hemorrhages in experimental settings and tested whether the addition of neuroprotective medication augments the efficacy of thrombolysis and reduces the incidence of hemorrhages. METHODS: Male Wistar rats were subjected to right middle cerebral artery embolization with an autologous thrombus and were then randomly assigned to one of the following groups: Group 1, saline-treated (2 hours after ischemic insult) animals as controls; Groups 2 to 4, high-dose urokinase (5,000 U/kg) at 2, 3, and 6 hours after the insult; Group 5, low-dose urokinase (2,500 U/kg) at 2 hours after the insult; Group 6, 20 mg/kg topiramate (TPM) at 2 hours after the insult; Group 7, a combination of 20 mg/kg TPM at 2 hours and low-dose urokinase (2,500 U/kg) at 6 hours after the insult; and Group 8, 20 mg/kg TPM (20 mg/kg) at 2 hours and high-dose urokinase (5,000 U/kg) at 2 hours after the insult. Neurological behavior and the infarct volume in the brain were assessed following cerebral embolism and the various treatments. All animals in the single therapy and low-dose combination groups survived surgery. Three of eight animals treated with high-dose urokinase alone at 6 hours and three of six animals in the combined high-dose urokinase and TPM group developed fatal intracerebral hemorrhages. There was a significantly better neurological outcome at 24 hours in the animals treated with either medication compared with controls. The volume of the infarct in the saline-treated group was 54.2 +/- 9%. The use of TPM at 2 hours led to a decrease in the infarct to 20.1 +/- 11.2% (p < 0.01). Treatment with urokinase at 6 hours after the occlusion showed a trend toward protection; the infarct volume was 31.9 +/- 14.1% (p < 0.05). The addition of TPM to low- or high-dose urokinase achieved better neuroprotection (8.2 +/- 6% and 11.9 +/- 10.7%, respectively; both p < 0.01). CONCLUSIONS: In this study the authors show that the volume of the infarct can be significantly decreased with 2 to 6-hour delayed intraarterial thrombolysis with urokinase and that the efficacy of thrombolysis may be enhanced by combining neuroprotective agents like TPM. It is also shown that low-dose combination therapy may decrease the likelihood of cerebral hemorrhage. PMID- 10794302 TI - Marinacci communication. Case report. AB - Marinacci or reverse Martin-Gruber communication is a rare anatomical variation. Until now, in all reported cases the branch was diagnosed using electrophysiological studies but not substantiated anatomically. The authors report a case of ulnar-median nerve communication in the distal forearm that was discovered during carpal tunnel release surgery and confirmed postoperatively on electromyographic studies. PMID- 10794301 TI - Significant reduction in brain swelling by administration of nonpeptide kinin B2 receptor antagonist LF 16-0687Ms after controlled cortical impact injury in rats. AB - OBJECT: Identification of new therapeutic agents aimed at attenuating posttraumatic brain edema formation remains an unresolved challenge. Among others, activation of bradykinin B2 receptors is known to mediate the formation of brain edema. The purpose of this study was to investigate the protective effect of the novel nonpeptide B2 receptor antagonist, LF 16-0687Ms, in brain injured rats. METHODS: Focal contusion was produced by controlled cortical impact injury. Five minutes after trauma, the rats received a single dose of no, low- (3 mg/kg body weight), or high- (30 mg/kg) dose LF 16-0687Ms. After 24 hours, the amount of brain swelling and hemispheric water content were determined. Low and high doses of LF 16-0687Ms significantly reduced brain swelling by 25% and 27%, respectively (p < 0.03). Hemispheric water content tended to be increased in the nontraumatized hemisphere. In a subsequent series of 10 rats, cisternal cerebrospinal fluid (CSF) samples were collected to determine whether changes in substances associated with edema formation could clarify why LF 16-0687Ms increases water content. For this, the volume regulator amino acid taurine, the excitatory transmitter glutamate, and the adenosine triphosphate degradation products hypoxanthine and xanthine were measured. In CSF, the levels of taurine, hypoxanthine, and xanthine were significantly decreased following a single administration of LF 16-0687Ms (p < 0.005); the level of glutamate, however, was double that found in control animals (p < 0.05). CONCLUSIONS: Using the present study design, a single administration of LF 16-0687Ms successfully reduced posttraumatic brain swelling. The decreased levels of taurine, hypoxanthine, and xanthine may reflect reduced posttraumatic brain edema, whereas the increased level of glutamate could account for the elevated water content observed in the nontraumatized hemisphere. PMID- 10794303 TI - Enhancement of survival of stored dopaminergic cells and promotion of graft survival by exposure of human fetal nigral tissue to glial cell line--derived neurotrophic factor in patients with Parkinson's disease. Report of two cases and technical considerations. AB - The authors have studied the ability of glial cell line-derived neurotrophic factor (GDNF) to promote survival of human fetal dopaminergic tissue after a storage period of 6 days and subsequent implantation into the human putamen. The results indicate that GDNF promotes survival of stored dopaminergic cells. Cells stored without GDNF had a 30.1% decrease in survival time compared with those exposed to GDNF. Two patients with Parkinson's disease received bilateral putaminal implants of fetal dopaminergic cells exposed to GDNF for 6 days and showed enhancement of graft survival as assessed by positron emission tomography scanning. A mean increase of 107% in putaminal fluorodopa uptake from baseline values was observed 12 months postgrafting. PMID- 10794305 TI - Diagnosis of spontaneous intracranial hypotension by using magnetic resonance myelography. Case report. AB - The authors describe a case of spontaneous intracranial hypotension in which the leakage site was determined by using magnetic resonance (MR) myelography. This technique demonstrated the route of cerebrospinal fluid (CSF) leakage, whereas other methods failed to show direct evidence of leakage. Magnetic resonance myelography is a noninvasive method that is highly sensitive in detecting CSF leakage. This is the first report in which a site of CSF leakage was detected using MR myelography. PMID- 10794304 TI - Adult choroid plexus papilloma of the posterior fossa mimicking a hemangioblastoma. Case report. AB - The authors report a case of cystic choroid plexus papilloma that originated in the posterior fossa. No connection with the ventricular system was found intraoperatively. Magnetic resonance (MR) and computerized tomography imaging did not furnish a diagnosis, but findings of pathological examinations were consistent with those of choroid plexus papilloma. The authors describe the different appearances of the tumor on MR images and discuss the differential diagnosis with other tumors of the posterior fossa. PMID- 10794306 TI - Subfascial and submuscular methods of temporal muscle dissection and their relationship to the frontal branch of the facial nerve. Technical note. AB - The microsurgical anatomy of the temporal and zygomatic branches of the facial nerve are presented along with related local vasculature (frontal and parietal branches of the superficial temporal artery [STA]) as encountered when using subfascial and submuscular temporal muscle dissection techniques for anterolateral craniotomies. Twenty sides were studied in 10 cadaveric specimens that had been previously injected with latex. The rami of the temporal and zygomatic branches of the facial nerve and branches of the STA were dissected out through pterional and orbitozygomatic approaches by using a submuscular or subfascial temporal muscle dissection technique. The three rami of the temporal branch of the facial nerve (the auricularis, frontalis, and orbicularis) were found to run within the galeal plane of the scalp. The zygomatic branch of the facial nerve was found to course deeper than the most caudal extension of the galea, known as the superficial musculoaponeurotic layer. The frontal branch of the STA served as an important landmark for the subfascial or submuscular dissections because excessive reflection of the scalp flap inferior to the level of this vessel would inadvertently injure the frontalis branch of the facial nerve. Subfascial and submuscular dissections of the temporal muscle offer an alternative to the interfascial technique during anterolateral craniotomies. Scalp and temporal dissection performed with careful attention to anatomical landmarks (frontal branch of the STA and the suprafascial fat pad) provides a safe and expeditious alternative to the traditional interfascial technique. PMID- 10794308 TI - Mathematical theory of stereotactic coordinate transformation: elimination of rotational targeting error by addition of a third reference point. Technical note. AB - All frame-based stereotactic procedures require localization of an anatomical target within the coordinate system of the stereotactic frame. If the target is defined by its coordinates given in a stereotactic atlas (indirect localization), the neurosurgeon faces the mathematical task of transforming atlas coordinates into frame coordinates. In the method usually used, the frame coordinates of two reference points (the anterior and posterior commissures) are obtained from computerized tomography or magnetic resonance images, and serve as the basis for the coordinate transformation. This two-point algorithm relies on the additional assumption that the frame sits on the patient's head without exhibiting roll, that is, rotation about the anteroposterior axis (y axis). Usually this assumption is nearly, but not exactly, correct. An amount of roll as small as 3 degrees can cause a targeting error on the order of 1 mm when a two-point algorithm is used. This potential source of error can be eliminated by using a new method of coordinate transformation, the derivation of which is mathematically reported in this article. The new method requires a third reference point located in the midsagittal plane, in addition to the two commissural reference points. PMID- 10794307 TI - Use of gadolinium as an intraarterial contrast agent in digital subtraction angiography of the cervical carotid arteries and intracranial circulation. Technical note. AB - Patients with renal insufficiency or other contraindications to iodine-based contrast agents present a significant management dilemma when conventional arteriography is required. The authors describe the use of gadolinium as an alternative contrast agent for arterial digital subtraction (DS) angiography of the cervical carotid arteries (CAs) and intracranial circulation. Three patients with renal insufficiency presented with symptoms of ischemic cerebrovascular disease and inconclusive noninvasive imaging studies, which necessitated conventional angiography. Traditional transfemoral catheterization of the cervical CAs was performed and DS angiographic studies were obtained using gadolinium as an intraarterial contrast agent. In one case, selective arteriographic examination of the internal carotid arteries and vertebrobasilar system was performed as well. High-quality, diagnostic images essentially indistinguishable from routine angiographic studies were obtained in all cases. No patient suffered a complication related to the use of gadolinium, and no patient demonstrated worsened renal function after the procedure. In the setting of a contraindication to iodine-based contrast agents, gadolinium represents an important alternative contrast material that allows for excellent visualization of the cervical CAs and intracranial circulation. PMID- 10794309 TI - Trigeminal neuralgia due to cerebellopontine angle lipoma. Case illustration. PMID- 10794310 TI - Isolated Rosai-Dorfman disease of the fourth ventricle. Case illustration. PMID- 10794311 TI - Gamma surgery for vestibular schwannoma. PMID- 10794312 TI - Gamma surgery for vestibular schwannoma. PMID- 10794313 TI - Magnetic resonance imaging and brainstem injury. PMID- 10794314 TI - Meningioma recurrence. PMID- 10794315 TI - Oxyhemoglobin and apoptosis. PMID- 10794316 TI - Extradural posterior inferior cerebellar artery. PMID- 10794317 TI - Colloid cysts. PMID- 10794318 TI - Colloid cysts. PMID- 10794319 TI - Colloid cysts. PMID- 10794320 TI - Moyamoya disease and transforming growth factor-beta1. PMID- 10794321 TI - Pupils and coma. PMID- 10794322 TI - Pineal epidermoid cysts. PMID- 10794323 TI - Apology. PMID- 10794324 TI - The role of the pulmonary artery catheter in the year 2000 and beyond. PMID- 10794325 TI - Clinical and economic effects of pulmonary artery catheterization in nonemergent coronary artery bypass graft surgery. AB - OBJECTIVE: To examine the association between use of pulmonary artery catheterization with hospital outcomes and costs in nonemergent coronary artery bypass graft (CABG) surgery. DESIGN: Retrospective cohort study. SETTING: Fifty six community-based hospitals in 26 states. PARTICIPANTS: A total of 13,907 patients undergoing nonemergent CABG surgery between January 1, 1997, and December 31, 1997. MEASUREMENTS AND MAIN RESULTS: Discharge abstracts for each patient were examined. Stratified and multivariate analyses were used to assess the impact of pulmonary artery catheters (PACs) on in-hospital mortality, length of stay in the intensive care unit, total length of stay, and hospital costs. Outcomes were adjusted for patient demographic factors, hospital characteristics, and hospital volume of PAC use in the year of analysis. Fifty-eight percent of the patients received a PAC. After adjustment, the relative risk of in-hospital mortality was 2.10 for the PAC group compared with the patients who did not receive a PAC (95% confidence interval [CI], 1.40 to 3.14; p < 0.001). The mortality risk was significantly higher in hospitals with the lowest third of PAC use (odds ratio, 3.35; 95% CI, 1.74 to 6.47; p < 0.001) and not significantly increased in the highest two thirds of users (odds ratio, 1.62; 95% CI, 0.99 to 2.66; p = 0.09). Days spent in critical care were similar; however, total length of hospital stay was 0.26 days longer in the PAC group (p < 0.001). Hospital costs were $1,402 higher in the PAC group. CONCLUSION: In the setting of nonemergent CABG surgery, pulmonary artery catheterization was associated with an increased risk of in-hospital mortality, greater length of stay, and higher total costs, particularly in hospitals with low volume of PAC use. PMID- 10794326 TI - Is the placement of a pulmonary artery catheter still justified solely for the measurement of cardiac output? AB - OBJECTIVE: The authors compared four clinical techniques of measuring cardiac output (CO) in critically ill patients: pulmonary artery thermodilution (CO[PA]), transpulmonary aortic thermodilution (CO[AORTA]), Fick principle-derived (CO[FICK]), and continuous pulmonary artery (CCO) measurements. DESIGN: Prospective clinical study. SETTING: Surgical intensive care unit of a university hospital. PARTICIPANTS: Twelve adult patients suffering from sepsis or septic shock. INTERVENTIONS: All patients were deeply sedated and mechanically ventilated in a pressure-controlled mode. Each patient received a 7.5F five-lumen pulmonary artery catheter for the continuous measurement of cardiac output and a 4F aortic catheter with an integrated thermistor. The thermistors of the two different catheters were connected to one computer system (COLD-Z021, Pulsion Medical Systems, Munich, Germany). Whole-body oxygen consumption was measured by indirect calorimetry using a metabolic cart (Deltatrac, Datex-Engstroem, Helsinki, Finland) over a 5-minute period, at the end of which arterial and mixed venous blood gases were taken and measured by co-oximetry. During each measuring period, three bolus CO measurements were performed. A total number of 51 CO measurements was analyzed. RESULTS: Linear regression analysis revealed the highest correlation between CO(AORTA) and CO(PA) (r = 0.98), whereas agreement between these two techniques and CCO was lower (r = 0.92 and r = 0.93). All three techniques correlated comparably with CO(FICK) (r = 0.85, r = 0.83, and r = 0.83). CONCLUSION: The correlations among the four CO techniques were high and similar, with CO(PA) and CO(AORTA) techniques showing the highest agreement. Because CO with similar accuracy can be obtained from transpulmonary aortic thermodilution in a less-invasive manner, it appears that the placement of a pulmonary artery catheter solely for the measurement of CO is no longer justified, unless continuous CO measurements are needed. PMID- 10794327 TI - Beat-to-beat measurement of cardiac output by intravascular pulse contour analysis: a prospective criterion standard study in patients after cardiac surgery. AB - OBJECTIVE: To evaluate the accuracy of a new pulse contour method of measuring cardiac output in critically ill patients. DESIGN: A prospective criterion standard study. SETTING: Cardiac surgery intensive care unit in a university hospital. PARTICIPANTS: Nineteen cardiac surgery patients requiring intensive care treatment with pulmonary artery catheters after surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The pulse contour cardiac output monitor uses transpulmonary bolus thermodilution measurements to calibrate the system. In each patient, the pulse contour cardiac output values were compared with conventional thermodilution. The method described by Bland and Altman and linear regression analysis were used for comparison. The mean difference (bias) +/- standard deviation of differences (precision) was 0.31 +/- 1.25 L/min for pulmonary bolus thermodilution cardiac output versus pulse contour cardiac output and 0.21 +/- 0.73 L/min for pulmonary bolus thermodilution cardiac output versus transpulmonary bolus thermodilution cardiac output. Linear regression (correlation) analyses were pulse contour cardiac output = 0.97 thermodilution + 0.53 (r = 0.88), and transpulmonary cardiac output = 0.87 thermodilution + 1.09 (r = 0.96). There was a small increase 60 minutes after recalibration but not a statistically significant difference between pulse contour cardiac output and pulmonary bolus thermodilution cardiac output (p = 0.52). CONCLUSIONS: Bias and precision are acceptable, and the system provides results that agree with conventional thermodilution. This study demonstrates the clinical applicability of the pulse contour cardiac output monitoring system. PMID- 10794328 TI - Transesophageal echocardiography values for left ventricular end-diastolic area and pulmonary vein and mitral inflow Doppler velocities in patients undergoing coronary artery bypass graft surgery. AB - OBJECTIVE: To determine left ventricular end-diastolic area (EDA) and pulmonary vein and mitral inflow Doppler velocities in patients undergoing coronary artery bypass graft (CABG) surgery using transesophageal echocardiography (TEE). To examine the effects of age, sex, and left ventricular function on these values. DESIGN: Prospective observational study; all measurements performed before cardiopulmonary bypass. SETTING: Tertiary referral teaching hospital. PARTICIPANTS: Eighty-six elective CABG surgery patients. INTERVENTIONS: Intraoperative TEE was performed in all patients. MEASUREMENTS AND MAIN RESULTS: The left ventricular EDA was measured at the midpapillary level, excluding the papillary muscles. Mean EDA for patients with normal left ventricular function, defined by fractional area change (FAC) 0.50 or greater, was 10.66 cm2 and when indexed to body surface area was 5.6 cm2/m2. The EDA was greater in patients with impaired left ventricular function (FAC < 0.50). Mean EDA was 14.84 cm2, and EDA/body surface area was 7.8 cm2/m2. In patients with FAC 0.50 or greater, mean peak pulmonary vein Doppler velocities were 46.10 cm/sec (systole), 31.71 cm/sec (diastole), and 1.50 (ratio systole to diastole). Mean peak mitral inflow Doppler velocities were 57.25 cm/sec (early diastole), 57.21 cm/sec (late diastole), and 1.10 (ratio early to late), and deceleration time was 216 msec. Age, sex, and left ventricular function were not significant predictors of Doppler variables. CONCLUSIONS: Values are presented for a predominantly white population undergoing cardiac surgery. Left ventricular dysfunction is associated with increased left ventricular EDA measurements. PMID- 10794329 TI - The effects of transesophageal echocardiography on hemodynamic variables in small infants undergoing cardiac surgery. AB - OBJECTIVE: To assess the effects of transesophageal echocardiography (TEE) on hemodynamic variables during cardiac surgery in small infants. DESIGN: A prospective clinical study. SETTING: A medical college-affiliated tertiary care children's hospital. PARTICIPANTS: Twenty-three infants weighing 2 to 5 kg undergoing cardiac surgery. INTERVENTIONS: Baseline heart rate, arterial pressure, and central venous pressure were recorded. A pediatric TEE probe was inserted, and the hemodynamic variables were again recorded. Postoperatively the hemodynamic measurements were measured again before and after probe removal, with the addition of left atrial pressure and pulmonary artery pressure when available. Hemodynamic parameters were carefully observed during all phases of the TEE examinations for any changes attributable to probe manipulation. MEASUREMENTS AND MAIN RESULTS: No statistically significant changes occurred in this group of patients during TEE. No clinically significant changes in any individual patient occurred during the measurement or during manipulation of the TEE probe for the complete examination. CONCLUSION: Although hemodynamic compromise can occur in small infants, this study suggests that it is infrequent. Fear of hemodynamic compromise should not prevent use of intraoperative TEE in small infants when otherwise indicated. PMID- 10794330 TI - Three-dimensional reconstructions of normal and aneurysmatic left ventricles in vivo using transesophageal echocardiography. AB - OBJECTIVE: To perform three-dimensional surface reconstructions to provide spatial delineations of a normal and an aneurysmatic left ventricle, using transesophageal echocardiography. DESIGN: Prospective study. SETTING: University hospital. PARTICIPANTS: Eight patients in cardiogenic shock admitted to the intensive care unit and two patients undergoing surgery with general anesthesia. INTERVENTIONS: Using a multiplane transesophageal echocardiography probe, nine echocardiographic cross-sectional images of the heart at approximately 20 degrees angular increments were obtained from midesophageal level in each patient for three-dimensional surface reconstructions. Multiple determinations of cardiac output using the thermodilution principle were also made in each patient to verify the accuracy of three-dimensional data sets. MEASUREMENTS AND MAIN RESULTS: End-diastolic and end-systolic volumes were determined from three dimensional data sets using the disc-summation method. Stroke volume was derived as the difference between end-diastolic and end-systolic volumes. Stroke volume was also calculated from thermodilution cardiac output measurements and heart rate. Correlation and limits of agreement between stroke volumes derived by the two methods were determined. Three-dimensional wire-frame models of a normal and an aneurysmatic left ventricle at end-systole were constructed from the nine echocardiographic cross-sectional images. Correlation coefficient between stroke volume derived from three-dimensional data sets using the disc-summation method and that measured by the thermodilution method was 0.91 (p < 0.001). Wire-frame models reveal a normal symmetric cavity and an aneurysmal cavity in sharp relief. CONCLUSIONS: Three-dimensional surface reconstruction can be performed from multiple cross-sectional images obtained using an unmodified commercially available multiplane transesophageal echocardiography probe, to reveal the left ventricular cavity in sharp relief. High correlation between stroke volume calculated from three-dimensional data sets and that measured by the thermodilution method attests to the accuracy of the three-dimensional data sets. PMID- 10794331 TI - Perioperative alterations in plasma endothelin-1 and echocardiographic correlates of right heart function. AB - OBJECTIVE: To determine whether greater changes in plasma endothelin-1 (ET-1) concentrations and right ventricular systolic pressure occur after major thoracic surgery than after major abdominal operations. DESIGN: Prospective study. SETTING: University hospital. PARTICIPANTS: Patients undergoing elective thoracotomies (n = 12) or laparotomies (n = 10). INTERVENTIONS: ET-1 was measured from blood obtained before anesthesia and again on postoperative days 1, 2, 3, and 5 (or 6). Transthoracic echocardiography was performed before surgery and on postoperative day 2 to evaluate right-sided heart function. MEASUREMENTS AND MAIN RESULTS: After abdominal and thoracic surgery, systemic and estimated pulmonary vascular pressures were normal in both groups and unaffected by surgery. Plasma ET-1 concentrations decreased from baseline values during the first postoperative week with no differences between the groups. CONCLUSIONS: In patients without organic heart disease, plasma ET-1 levels do not increase in response to major abdominal or thoracic surgery. Whether or not plasma ET-1 concentrations are elevated in patients developing clinically significant postoperative pulmonary hypertension requires further study. PMID- 10794332 TI - How silent is perioperative myocardial ischemia? A hemodynamic, electrocardiographic, and biochemical study in patients undergoing coronary artery bypass graft surgery. AB - OBJECTIVE: To analyze the relationship among Holter electrocardiogram (ECG) recordings, hemodynamic measurements indicative of global myocardial oxygen balance, and serum cardiac troponin I concentrations (cTnI) in the early postoperative period after coronary artery bypass graft (CABG) surgery. DESIGN: Prospective observational study. SETTING: University teaching hospital. PARTICIPANTS: Thirty patients undergoing CABG surgery. INTERVENTIONS: ECG measurements consisted of Holter and standard ECG recordings. Hemodynamic measurements included heart rate, systolic and diastolic blood pressure (SBP, DBP), pulmonary capillary wedge pressure, and cardiac index (CI). Derived indices included tension time index (TTI), rate-pressure product, pressure work index (PWI), and endocardial viability ratio (EVR). Serial measurements of cTnI concentrations were measured postoperatively; the area under the cTnI concentration time curve was calculated for each patient (AUC cTnI). MEASUREMENTS AND MAIN RESULTS: Episodes of myocardial ischemia were associated with small but significant rises in SBP (p = 0.01), DBP (p = 0.001), and TTI (p = 0.005) compared with periods without ischemia in the same patients. Serum cTnI concentrations 24 hours after cardiopulmonary bypass (p = 0.03) and AUCcTnI (p = 0.01) values were greater in patients who developed ECG myocardial ischemia compared with patients who did not. CONCLUSIONS: The small changes in hemodynamics seen, although statistically significant, are unlikely to be the primary cause of the ischemia. They more likely reflect an independent process that causes or occurs as a result of ischemic episodes. Ischemic episodes detected by the Holter monitor are associated with significant release of cardiac troponin from the myocardium. PMID- 10794333 TI - Minor cardiac troponin T release in patients undergoing coronary artery bypass graft surgery on a beating heart. AB - OBJECTIVES: To determine whether and to what extent coronary artery bypass graft (CABG) surgery without extracorporeal circulation is associated with cardiac troponin T (TnT) release. DESIGN: Prospective study. SETTING: A single university hospital. PARTICIPANTS: Twenty-three patients scheduled for minimally invasive CABG surgery. Sixteen patients received one coronary anastomosis, and seven received two. INTERVENTIONS: TnT and creatine kinase-MB (CK-MB) levels were determined immediately before induction of anesthesia (baseline) and at 0, 12, and 24 hours after surgery. Hemodynamic measurements were made, and 5-lead electrocardiograms with continuous automated ST-segment trends were analyzed. MEASUREMENTS AND MAIN RESULTS: All patients had a good cardiac outcome. Median cumulative coronary artery occlusion time was 27 minutes (range, 10 to 49 minutes). TnT levels were undetectable in 91.3% of patients at baseline when a detection limit of 0.01 ng/mL was employed. TnT and CK-MB showed significant elevations at 12 and 24 hours versus baseline. Postoperatively, TnT was detectable in 91.3% of patients, and 17.4% suffered minor myocardial damage, as evidenced by an abnormal increase in TnT greater than 0.2 ng/mL, excluding those exhibiting myocardial infarction. ST segment changes developed in seven patients, persisting for 13.0 minutes (range, 9.5 to 15.8 minutes) and disappearing immediately after coronary artery clamp release. There were no significant correlations between cumulative coronary occlusion time and peak TnT or CK-MB levels. CONCLUSIONS: TnT was detected after surgery in most patients, and significant TnT levels indicative of myocardial injury (>0.2 ng/mL) were detected in only 17% of patients, probably as a result of brief periods of coronary artery occlusion. PMID- 10794334 TI - Plasma interleukin-4, interleukin-10, and interleukin-13 concentrations and complications after coronary artery bypass graft surgery. AB - OBJECTIVES: To determine whether plasma interleukin (IL)-4, IL-10, and IL-13 concentrations are associated with complications after coronary artery bypass graft (CABG) surgery with cardiopulmonary bypass (CPB). STUDY DESIGN: Prospective descriptive study. SETTING: University teaching hospital. PARTICIPANTS: Thirty two patients during and 24 hours after CABG surgery. MEASUREMENTS AND MAIN RESULTS: Hemodynamic measurements and blood samples were obtained from 32 patients during and after surgery. Coagulation, pulmonary, and cardiovascular functions were specifically assessed during the first 24 hours postoperatively. Plasma IL-4 and IL-13 levels remained unchanged during CABG surgery. In contrast, plasma IL-10 concentrations increased by 117-fold in the immediate postoperative period and returned to pre-CPB values by 24 hours postoperatively. Plasma IL-10 levels were not different in patients with or without cardiovascular impairment, coagulation disorders, and lung injury. Plasma IL-10 levels did not correlate with the leukocyte count, the amount of catecholamines infused, or the duration of CPB. CONCLUSION: The present results suggest that the development of post-CABG surgery complications might be linked to an insufficient production of anti inflammatory cytokines, such as IL-4, IL-10, or IL-13, which are unable to counteract the overproduction of inflammatory cytokines. PMID- 10794335 TI - Reactive hyperemia in skin after cardiopulmonary bypass. AB - OBJECTIVE: To study reactive hyperemia (RH) using a transcutaneous PO2/PCO2 combination electrode heated to 37 degrees C and tissue reflectance spectrophotometry in patients before and after cardiopulmonary bypass (CPB) to determine whether microcirculatory function of skin is altered. DESIGN: Prospective study. SETTING: Anesthesiology and critical care unit of a university hospital. PARTICIPANTS: Eight patients undergoing elective CPB under mild hypothermia. INTERVENTIONS: To produce RH, blood flow to the forearm was prevented by inflation of a cuff to 300 mmHg for an interval of 5 minutes. MEASUREMENTS AND MAIN RESULTS: Measurements were obtained on the day prior to surgery (DPS), on the day of surgery (DOS) rewarmed to 37 degrees C in the intensive care unit (ICU), and on the first (POD 1) and the third postoperative days (POD 3). The following parameters were recorded: preocclusive baseline cutaneous PO2, and PCO2 (B-PtcO2, B-PtcCO2), and microvascular hemoglobin saturation (B-HbO2); postischemic peak of PtcO2, PtcCO2, and HbO2; and 10 minutes after release of the cuff occlusion posthyperemic PtcO2, PtcCO2, and HbO2. B PtcO2 was 3.5 +/- 1.2 mmHg on DPS, 2.6 +/- 0.7 mmHg on DOS, 1.5 +/- 0.3 mmHg on POD 1, and 3.5 +/- 3.5 mmHg on POD 3. B-PtcCO2 increased significantly from 40.1 +/- 2.5 mmHg to 52.2 +/- 2.0 mmHg on DOS (p = 0.01) and to 48.9 +/- 3.6 mmHg on POD 1 (p = 0.02). On POD 3, B-PtcCO2 was 40.6 +/- 2.6 mmHg. B-HbO2 declined from a preoperative value of 42.4% +/- 8.6% to 37.1% +/- 14.7% on DOS and further to 21.7% +/- 4.8% on POD 1, which was significantly different (p = 0.03). On POD 3, B-HbO2 still remained lower (30.7% +/- 6.2%) compared with the preoperative value. RH (deltaPtcO2, deltaHBO2) was quantified as the differences between peak PtcO2, HBO2 and B-PtcO2, B-HBO2. DeltaPtcO2 was 13.0 +/- 2.3 on DPS, 11.3 +/- 2.9 on DOS, 12.6 +/- 2.6 on POD 1, and 11.5 +/- 3.5 on POD 3. DeltaHBO2 was 42.0 +/- 5.6 on DPS, 40.0 +/- 7.1 on DOS, 49.9 +/- 2.5 on POD 1, and 52.9 +/- 6.4 on POD 3. The elimination rate of carbon dioxide from skin (ECO2) was calculated as difference between peak PtcCO2 and PtcCO2 after 3 minutes of reperfusion divided by the difference between peak PtcCO2 and B-PtcCO2. ECO2 was 1.0 +/- 0.2 kPa/min on DPS, 0.7 +/- 0.1 kPa/min on DOS, and 0.8 +/- 0.1 kPa/min on POD 1 and POD 3. CONCLUSION: Cutaneous microcirculation assessed by RH is well preserved during the immediate postoperative period in patients undergoing uncomplicated coronary artery surgery with CPB. PMID- 10794336 TI - Subtle brain damage cannot be detected by measuring neuron-specific enolase and S 100beta protein after carotid endarterectomy. AB - OBJECTIVE: To assess whether subtle brain damage after carotid endarterectomy could be detected using serum levels of neuron-specific enolase (NSE) or S 100beta protein. DESIGN: Prospective noninterventional study. SETTING: University hospital. PARTICIPANTS: Twenty-two patients undergoing carotid endarterectomy and 16 patients undergoing repair of abdominal aortic aneurysm. INTERVENTIONS: Serum levels of NSE and S-100beta protein were measured in all patients before surgery and postoperatively at 12, 24, 36, and 48 hours. In patients undergoing carotid endarterectomy, neuropsychologic testing was performed before surgery and postoperatively at discharge from the hospital and after 3 months using a neuropsychologic test battery. MEASUREMENTS AND MAIN RESULTS: Compared with abdominal aortic surgery patients, the preoperative serum concentration of NSE was significantly higher in carotid artery surgery patients. Postoperatively, the NSE serum level decreased significantly after uncomplicated carotid artery surgery, and the level was then similar to that in the aortic surgery patients. Before operation, the S-100beta protein levels were similar in the two groups, but a significant increase was seen in aortic surgery patients postoperatively. Neuropsychologic testing after uncomplicated carotid artery surgery revealed cognitive dysfunction in 2 of 20 (10%) of the patients after 5 days and 3 of 16 (19%) of the patients after 3 months. There was no correlation between the change in cognitive function and the changes in blood levels of either NSE or S-100 protein. CONCLUSIONS: Subtle brain damage after carotid artery surgery could not be detected by measuring blood levels of NSE and S-100beta protein. The NSE level was significantly higher before carotid artery surgery and decreased postoperatively to the level observed in aortic surgery. PMID- 10794337 TI - Institutional variability in red blood cell conservation practices for coronary artery bypass graft surgery. Institutions of the MultiCenter Study of Perioperative Ischemia Research Group. AB - OBJECTIVE: To assess whether substantial institutional variability exists in red blood cell conservation practices associated with coronary artery bypass graft (CABG) surgery. DESIGN: Prospective, randomized patient enrollment and data collection. SETTING: Twenty-four U.S. academic institutions participating in the Multicenter Study of Perioperative Ischemia. PARTICIPANTS: A well-defined subset of primary CABG surgery patients (n = 713) expected to be at low risk for bleeding and exposure to allogeneic transfusion. INTERVENTIONS: None (observational study). MEASUREMENTS AND MAIN RESULTS: Frequency of use of red blood cell conservation techniques was determined among institutions. Correlation was determined between use of each technique and transfusion of allogeneic red blood cells and between use of each technique and median institutional blood loss. Significant variability (p < 0.01) was detected in institutional transfusion practice with respect to the use of predonated autologous whole blood, normovolemic hemodilution, red cell salvage, and reinfusion of shed mediastinal blood. The frequency of institutional use of these techniques was not associated with allogeneic transfusion (r2 < 0.15) or blood loss (r2 < 0.10) in the low-risk population of patients examined. CONCLUSIONS: Institutions vary significantly in perioperative blood conservation practices for CABG surgery. Further study to determine the appropriate use of these techniques is warranted. PMID- 10794339 TI - Re-evaluation of renal cell carcinoma tumor thrombus extension by intraoperative transesophageal echocardiography. PMID- 10794338 TI - Blood loss from coronary angiography increases transfusion requirements for coronary artery bypass graft surgery. AB - OBJECTIVE: To determine the blood loss associated with coronary angiography and its impact on hemoglobin and transfusion requirements for subsequent coronary artery bypass graft (CABG) surgery. DESIGN: Retrospective chart review. SETTING: Tertiary-care, academic medical center. PARTICIPANTS: A total of 506 adult patients undergoing coronary angiography and CABG surgery. INTERVENTIONS: None (observational study). MEASUREMENTS AND MAIN RESULTS: Coronary angiography was associated with a reduction in hemoglobin of 1.8 g/dL. This reduction in hemoglobin was a significant predictor of allogeneic red blood cell transfusion. CONCLUSION: Coronary angiography contributes to a 1.8 g/dL reduction in hemoglobin concentration before CABG surgery and was associated with increased transfusion of allogeneic blood products. Measures aimed at maintaining red cell volume during coronary angiography, increasing erythropoiesis, or delaying surgery beyond 2 weeks may result in a decrease in transfusion requirements for patients undergoing CABG surgery. PMID- 10794340 TI - Airway obstruction by a transesophageal echocardiography probe in an adult patient with a dissecting aneurysm of the ascending aorta and arch. PMID- 10794341 TI - Cardiac operation in a patient with combined homozygous protein C and protein S deficiency. PMID- 10794342 TI - Differences in brain oxygenation between two sequential cardiac arrests during axillobifemoral bypass in a Stanford type A dissecting aneurysm. PMID- 10794343 TI - Magnesium therapy for refractory ventricular fibrillation. PMID- 10794344 TI - Propofol-induced anaphylactoid reaction during anesthesia for cardiac surgery. PMID- 10794345 TI - Preoperative assessment for pulmonary resection. PMID- 10794346 TI - Case 2-2000. Transesophageal echocardiography-associated gastrointestinal trauma. PMID- 10794347 TI - Pro: Preoperative thallium testing should be routinely performed before vascular surgery. PMID- 10794348 TI - Con: Preoperative thallium testing should not be performed routinely before vascular surgery. PMID- 10794349 TI - An unexpected echocardiographic image in a patient with dyspnea. PMID- 10794350 TI - An interesting finding during transesophageal echocardiographic assessment of the mitral valve. PMID- 10794351 TI - Alternative methods of inserting a new endobronchial blocker. PMID- 10794352 TI - Hemodynamic changes during off-pump coronary artery bypass graft surgery. PMID- 10794353 TI - Embracing the heart: perioperative management of patients undergoing off-pump coronary artery bypass grafting using the octopus tissue stabilizer. PMID- 10794354 TI - Arrayed primer extension: solid-phase four-color DNA resequencing and mutation detection technology. AB - The technology and application of arrayed primer extension (APEX) is presented. We describe an integrated system with DNA chip and template preparation, multiplex primer extension on the array, fluorescence imaging, and data analysis. The method is based upon an array of oligonucleotides, immobilized via the 5' end on a glass surface. A patient DNA is amplified by PCR, digested enzymatically, and annealed to the immobilized primers, which promote sites for template dependent DNA polymerase extension reactions using four unique fluorescently labeled dideoxy nucleotides. A mutation is detected by a change in the color code of the primer sites. The technology was applied to the analysis of 10 common beta thalassemia mutations. Nine patient DNA samples, each of which carries a different mutation, and four wild-type DNA samples were correctly identified. The signal-to-noise ratio of this technology is, on the average, 40:1, which enables the identification of heterozygous mutations with a high confidence level. The APEX method can be applied to any DNA target for efficient analysis of mutations and polymorphisms. PMID- 10794355 TI - Sensitivity of multiple color spectral karyotyping in detecting small interchromosomal rearrangements. AB - Multiple color spectral karyotyping (SKY) has been proven to be a very useful tool for characterization of the complex rearrangements in cancer cells and the de novo constitutional structural abnormalities. The sensitivity of SKY in detecting interchromosomal alterations was assessed with 10 constitutional translocations involving subtelomeric regions. Among the 13 small segments tested, 9 were clearly visualized and 8 were unambiguously identified by SKY. Fluorescence in situ hybridizations (FISH) with subtelomeric probes confirmed the reciprocity in three of the four translocations in which a small segment was not detectable by SKY. On the basis of resolution level of G-banding and the information obtained from the FISH analysis, the minimum alteration that SKY can detect is estimated to be 1,000-2,000 kbp in size with the currently available probes. This study has demonstrated the power, but also the limitations, of SKY in detecting small interchromosomal alterations, particularly those in subtelomeric regions. PMID- 10794356 TI - Efficient mutation detection in mismatch repair genes using a combination of single-strand conformational polymorphism and heteroduplex analysis at a controlled temperature. AB - Single-strand conformational polymorphism analysis (SSCP) and heteroduplex analysis (HD) were tested as methods for mutation screening with respect to experimental variation, sensitivity, and specificity. Thirty-nine fluorescently labeled PCR products covering the two mismatch repair genes, hMLH1 and hMSH2, were tested in 15 patients for pattern changes, using SSCP and HD at two temperatures, in a total of 2340 runs. SSCP was most efficient in detecting base changes, whereas HD was the method of choice when detecting deletions. SSCP and HD at 20 degrees C were most effective (sensitivity 97%, specificity 49%), and SSCP and HD at 10 degrees C gave no additional information, except in one case where an exon had two base changes. Several mutations only showed a small pattern change in one of the two strands, most explicit at 20 degrees C. No correlation between the type of base change and the size or direction of the pattern changes could be found. PMID- 10794357 TI - P53 mutations, polymorphisms, and haplotypes in Pakistani ethnic groups and breast cancer patients. AB - Inactivation of the p53 gene has been found to be associated with the pathogenesis of several neoplasias. Three biallelic polymorphisms in the p53 gene have been linked to predisposition to the development of various malignancies. These include a 16-bp duplication in intron 3 and BstU I and Msp I restriction fragment length polymorphisms (RFLPs) in exon 4 and intron 6, respectively. The prevalence of these polymorphisms was studied in breast cancer patients and nine major ethnic groups of Pakistan. Differences in allele frequencies for all three polymorphisms were observed among the various ethnic groups and breast cancer patients. The absence of the 16-bp duplication was common among the northern ethnic groups, being highest in the Hazara (0.90). The Msp I A1 allele frequency in the southern Makrani population was significantly higher in comparison with the other ethnic groups. In the cancer patients, the absence of the 16-bp duplication in combination with the BstU I Pro and absence of Msp I restriction site were the most frequent. In these patients, ten substitution mutations were found in the p53 gene, seven of which have been reported previously for breast cancer. The remaining three mutations have been found in other malignancies, but not in carcinoma of the breast. PMID- 10794358 TI - Impact of genetic privacy legislation on insurer behavior. PMID- 10794359 TI - Counseling by computer: breast cancer risk and genetic testing. Developed by the University of Wisconsin-Madison Department of Medicine and the Program in Medical Ethics. PMID- 10794360 TI - The predictive value of BRCA1 and BRCA2 mutation testing. AB - Genetic testing for mutations in BRCA1 and BRCA2, two genes predisposing to breast and ovarian cancers, is available to women with a relevant family history. The aim of this study was to estimate the positive and negative predictive value of clinical sequence analysis of these genes. A reference graph showing positive and negative predictive values over a range of pre-test risk was derived, taking into account the sensitivity and specificity of a full-sequence analysis test. High positive and negative predictive values were found for women with pre-test risk between 4% and 40%, a range of risk commonly seen in clinical testing. The predictive value of full sequence and single-site analysis of BRCA1 and BRCA2, therefore, compares favorably with other diagnostic medical tests. Our results provide a numerical estimate of the predictive value of BRCA testing, and as such, provide a valuable tool to healthcare providers and families as they interpret BRCA1 and BRCA2 test results. PMID- 10794361 TI - Exon deletions and duplications in BRCA1 detected by semiquantitative PCR. AB - rearrangements have recently been identified in the BRCA1 gene. Inclusion of a method for identifying such rearrangements should now be a prerequisite for providing a comprehensive mutation detection strategy. We have developed a semiquantitative PCR-based fluorescent assay for the detection of previously identified deletions. This method avoids the need for long PCR or Southern blotting and is suitable for large-scale epidemiological studies. The assay was used to screen 44 high-risk families within the U.K. Yorkshire Health Region. No deletions were detected, but five cases (11%) with an apparent duplication of exon 13 in BRCA1 were identified. The presence of this mutation was confirmed by long PCR. Further developments include extending the assay to include all exons of BRCA1. PMID- 10794362 TI - Null alleles at the Huntington disease locus: implications for diagnostics and CAG repeat instability. AB - PCR amplification of the CAG repeat in exon 1 of the IT15 gene is routinely undertaken to confirm a clinical diagnosis of Huntington disease (HD) and to provide predictive testing for at-risk relatives of affected individuals. Our studies have detected null alleles on the chromosome carrying the expanded repeat in three of 91 apparently unrelated HD families. Sequence analysis of these alleles has revealed the same mutation event, leading to the juxtaposition of uninterrupted CAG and CCG repeats. These data suggest that a mutation-prone region exists in the IT15 gene bounded by the CAG and CCG repeats and that caution should be exercised in designing primers that anneal to the region bounded by these repeats. Two of the HD families segregated null alleles with expanded uninterrupted CAG repeats at the lower end of the zone of reduced penetrance. The expanded repeats are meiotically unstable in these families, although this instability is within a small range of repeat lengths. The haplotypes of the disease-causing chromosomes in these two families differ, only one of which is similar to that reported previously as being specific for new HD mutations. Finally, no apparent mitotic instability of the uninterrupted CAG repeat was observed in the brain of one of the HD individuals. PMID- 10794363 TI - Rapid confirmation of previously detected prenatal mosaicism by fluorescence in situ hybridization in interphase uncultured amniocytes. AB - Fluorescence in situ hybridization (FISH) of chromosome-specific probes to interphase uncultured amniocytes was performed in cases in which follow-up amniocenteses were done for confirmation of previously detected mosaicism. FISH results were informative in all seven cases included in the study, and confirmed by subsequent cytogenetic analysis. FISH analysis provides rapid results for referral physicians and in most cases reassurance for patients within 24 hours of the follow-up aminocentesis. Although FISH studies are not considered accurate in determining a primary diagnosis of mosaicism in uncultured cells, the analysis is accurate and clinically useful when the diagnosis is known and mosaicism involving a specific chromosome needs to be confirmed in follow-up testing. PMID- 10794364 TI - Identification of the Hind III polymorphic site in the PAI-1 gene: analysis of the PAI-1 Hind III polymorphism by PCR. AB - The identification of the Hind III polymorphic site in the 3' end of the plasminogen activator inhibitor 1 (PAI-1) gene and a simple method to identify the Hind III polymorphism rapidly in the PAI-1 gene using PCR is described. The Hind III restriction site was identified by restriction site mapping and sequence analysis from a cosmid DNA clone. Genomic DNA was isolated from individual human umbilical cords and a 754-bp fragment of the human PAI-1 gene was amplified by PCR. Aliquots of the PCR products were digested with Hind III and analyzed by agarose gel electrophoresis. The presence of two fragments, 754 and 567 bp, was identified, and they were designated as 1/1 (750-bp band), 1/2 (754- and 567-bp bands), and 2/2 (567-bp band). The PCR method is considerably less time consuming than the conventional DNA genotyping using Southern blot analysis. To ensure that this new method identified the same PAI-1 genotypes as previously identified by Hind III restriction fragment length polymorphism (RFLP), samples were simultaneously genotyped by PCR and Southern blot analysis. Both methods identified the same Hind III genotypes in all the samples, confirming the reliability of this new PCR method for the rapid identification of the Hind III polymorphism in the human PAI-1 gene. PMID- 10794365 TI - Molecular analysis in Brazilian cystic fibrosis patients reveals five novel mutations. AB - We have performed molecular genetic analyses on 160 Brazilian patients diagnosed with cystic fibrosis (CF). Screening of mutations in 320 CF chromosomes was performed through single strand conformation polymorphism (SSCP) and heteroduplex analyses assay followed by DNA sequencing of the 27 exons and exon/intron boundaries of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The frequency of CFTR variants of T-tract length of intron 8 (IVS8 Tn) was also investigated. This analysis enabled the detection of 232/320 CF mutations (72.2%) and complete genotyping of 61% of the patients. The deltaF508 mutation was found in 48.4% of the alleles. Another fifteen mutations (previously reported) were detected: G542X, R1162X, N1303K, R334W, W1282X, G58E, L206W, R553X, 621+1G-->T, V232D, 1717-1G-->A, 2347 delG, R851L, 2789+5G-->A, and W1089X. Five novel mutations were identified, V201M (exon 6a), Y275X (exon 6b), 2686 insT (exon 14a), 3171 delC (exon 17a), and 3617 delGA (exon 19). These results contribute to the molecular characterization of CF in the Brazilian population. In addition, the identification of the novel mutation Y275X allowed prenatal diagnosis in a high-risk fetus. PMID- 10794366 TI - Evaluation of the prothrombin gene polymorphism in patients with advanced retinopathy of prematurity. AB - It has been reported recently that a common genetic variant in the 3' untranslated region of the prothrombin gene is associated with a significant fraction of premature births. The purpose of this study is to evaluate the prothrombin gene polymorphism in a large cohort of patients with preterm birth and advanced retinopathy of prematurity. For this purpose, the leukocyte DNAs were analyzed for the mutation (20210A) in the 3'-untranslated region of the prothrombin gene by PCR amplification, followed by restriction analysis and DNA sequencing. Our extensive analysis revealed a normal genotype (GG) in all patients as well as controls. These results suggest that the common genetic variant in the 3'-untranslated region of the prothrombin gene is not associated with advanced retinopathy of prematurity. Although more patients' samples should be evaluated, this genetic test does not support a relationship between prothrombin gene mutation and retinopathy of prematurity. PMID- 10794367 TI - Reflections on the European Conference "Molecular Screening of Individuals at High Risk for Developing Cancer: Medical, Ethical, Legal, and Social Issues". AB - The fascinating progress of molecular biology in the last decades has made possible the early identification of persons at risk of developing malignant neoplasms, such as thyroid, colon, and breast cancer. This has changed conventional medical practice and has raised a number of ethical questions. Despite the fact that there has been substantial development throughout the Western countries in legislation dealing with medical information, there is an acute need for further exploration and assessment of the moral and social dimensions of medical practices to achieve more precise and uniform regulations. These topics were discussed in a multidisciplinary European Conference entitled "Molecular Screening of Individuals at High Risk of Developing Cancer: Medical, Ethical, Legal, and Social Issues," which took place in March, 1999, in Athens, Greece. PMID- 10794368 TI - Chromosome 13. PMID- 10794369 TI - Alcohol myopia and condom use: can alcohol intoxication be associated with more prudent behavior? AB - We tested 2 competing theories about the effects of alcohol on intentions to engage in risky behavior. Disinhibition predicts that intoxicated people will exhibit risky behavior regardless of environmental cues, whereas alcohol myopia (C. M. Steele & R. A. Josephs, 1990) predicts that intoxicated people will be more or less likely to exhibit risky behavior, depending on the cues provided. In 4 studies, we found an interaction between intoxication and cue type. When impelling cues were present, intoxicated people reported greater intentions to have unprotected sex than did sober people. When subtle inhibiting cues were present, intoxicated and sober people reported equally cautious intentions (Studies 1-3). When strong inhibiting cues were present, intoxicated people reported more prudent intentions than did sober people (Study 4). We suggest that alcohol myopia provides a more comprehensive account of the effects of alcohol than does disinhibition. PMID- 10794370 TI - Bracing for loss. AB - People find unexpected bad news aversive and often brace themselves by predicting the worst. Three experiments examined whether the pessimism is influenced by personal need. Students who differed in financial need learned that a billing error meant that some students would receive an additional bill from their university. Financially needy students were consistently pessimistic in predicting their likelihood of receiving a bill, whereas non-needy students were not. In addition, the experiments reveal that (a) the pessimism occurred for potential losses but not potential gains, (b) needy students were pessimistic about their own chances but not the chances of a friend, (c) the pessimism was not attributable to needy students' being more readily primed by the news of a possible bill or to needy students' having more experience with billing errors, and (d) the pessimism was specific to monetary losses and did not generalize to other events. PMID- 10794371 TI - Framing interethnic ideology: effects of multicultural and color-blind perspectives on judgments of groups and individuals. AB - In 3 experiments, White American college students received a message advocating either a color-blind or a multicultural ideological approach to improving interethnic relations and then made judgments about various ethnic groups and individuals. Relative to a color-blind perspective, the multicultural perspective led to stronger stereotypes, greater accuracy in these stereotypes, and greater use of category information in judgments of individuals. This increase in between category differentiation occurred both for attributes that favored the in-group and for attributes that favored the out-group and was also paired in some cases with greater overall positivity toward the out-group. The findings lead us to question the implicit assumption driving the majority of social psychological efforts at prejudice reduction: that the categorization process leads to prejudice, and that the relevance of social categories must therefore be de emphasized. PMID- 10794372 TI - Biased information search in group decision making. AB - Research has shown that people prefer supporting to conflicting information when making decisions. Whether this biased information search also occurs in group decision making was examined in three experiments. Experiment 1 indicated that groups as well as individuals prefer supporting information and that the strength of this bias depends on the distribution of the group members' initial decision preferences. The more group members had chosen the same alternative prior to the group discussion (group homogeneity), the more strongly the group preferred information supporting that alternative. Experiment 2 replicated these results with managers. Experiment 3 showed that the differences between homogeneous and heterogeneous groups reflect group-level processes. Higher commitment and confidence in homogeneous groups mediated this effect. Functional and dysfunctional aspects of biased information seeking in group decision making are discussed. PMID- 10794373 TI - Displaced aggression is alive and well: a meta-analytic review. AB - Content analysis of 122 social psychology textbooks confirmed that displaced aggression received a surge of attention immediately following J. Dollard, L. W. Doob, N. E. Miller, O. H. Mowrer, and R. R. Sears (1939), but subsequent interest sharply declined. Contemporary texts give it little attention. By contrast, meta analysis of the experimental literature confirms that it is a robust effect (mean effect size = +0.54). Additionally, moderator analyses showed that: (a) The more negative the setting in which the participant and target interacted, the greater the magnitude of displaced aggression; (b) in accord with N. E. Miller's (1948) stimulus generalization principle, the more similar the provocateur and target, the more displaced aggression; and (c) consistent with the contrast effect (L. Berkowitz & D. A. Knurek, 1969), the intensity of initial provocation is inversely related to the magnitude of displaced aggression. PMID- 10794374 TI - Meta-stereotype activation: evidence from indirect measures for specific evaluative concerns experienced by members of dominant groups in intergroup interaction. AB - Six experiments demonstrated that dominant group members readily frame intergroup interaction in terms of how they themselves are evaluated. The authors used indirect measures of meta-stereotype activation to assess dominant group members' inclination to spontaneously consider an out-group member's (ostensible) stereotypic expectations about them. The necessary conditions for meta-stereotype activation were rather minimal, but the potential for evaluation by an out-group member--as opposed to mere exposure to the person--was required. Individual differences involving the importance accorded to social evaluation (public self consciousness and personal importance of racial attitudes) were associated with meta-stereotype activation, whereas racial attitudes were not. Two studies in which evaluative orientation was manipulated directly demonstrated a link between thinking in terms of how one is viewed and the activation and application of meta stereotypes. PMID- 10794375 TI - Perspective-taking: decreasing stereotype expression, stereotype accessibility, and in-group favoritism. AB - Using 3 experiments, the authors explored the role of perspective-taking in debiasing social thought. In the 1st 2 experiments, perspective-taking was contrasted with stereotype suppression as a possible strategy for achieving stereotype control. In Experiment 1, perspective-taking decreased stereotypic biases on both a conscious and a nonconscious task. In Experiment 2, perspective taking led to both decreased stereotyping and increased overlap between representations of the self and representations of the elderly, suggesting activation and application of the self-concept in judgments of the elderly. In Experiment 3, perspective-taking reduced evidence of in-group bias in the minimal group paradigm by increasing evaluations of the out-group. The role of self-other overlap in producing prosocial outcomes and the separation of the conscious, explicit effects from the nonconscious, implicit effects of perspective-taking are discussed. PMID- 10794376 TI - Interpersonal evaluations following threats to self: role of self-esteem. AB - In 2 studies, the authors used dyadic interactions to assess the influence of ego threat on likability as a function of self-esteem. In both studies, 2 naive participants engaged in a structured conversation; in half of the dyads, 1 participant received an ego threat prior to the interaction. In the 1st study, threatened high self-esteem participants were rated as less likable than were threatened low self-esteem participants. The 2nd study confirmed that ego threats are associated with decreased liking for those with high self-esteem and with increased liking for those with low self-esteem. A mediational analysis demonstrated that decreased liking among high self-esteem participants was due to being perceived as antagonistic. Study 2 also indicated that the findings could not be explained by trait levels of narcissism. These patterns are interpreted in terms of differential sensitivity to potential interpersonal rejection. PMID- 10794377 TI - Retrieving positive memories to regulate negative mood: consequences for mood congruent memory. AB - Several researchers have suggested that mood-incongruency effects are due to a mood-regulatory process in which people retrieve positive memories to repair negative moods. The present studies tested this idea by manipulating mood-repair strategies and examining their impact on positive and negative memory retrieval. Mood-congruent retrieval occurred when participants stayed focused on events associated with their negative mood; mood-incongruent retrieval occurred when participants engaged in positive reappraisal (when they reinterpreted events as having positive outcomes). The effects of these strategies on memory retrieval also interacted with personality traits related to negative mood regulation. Individuals high in such traits showed stronger mood-incongruent memory than did individuals low in negative mood-regulation traits. Discussion focuses on integrating mood-regulatory variables and personality variables into existing mood-congruency theories (e.g., associative network models). PMID- 10794378 TI - Don't mind if I do: disinhibited eating under cognitive load. AB - Past research has shown that strong emotional or motivational states can cause normally restrained eaters to overeat. In this article it is argued that simple cognitive load can also disinhibit eating by restrained eaters. Two studies examined this disinhibition effect. In Study 1, restrained and unrestrained eaters were given the opportunity to consume high-calorie food while performing either a high cognitive-load or low cognitive-load task. Restrained eaters consumed more food when under high cognitive load than when under low cognitive load; unrestrained eaters showed the opposite pattern. Study 2 replicated the disinhibition effect and ruled out stress, diminished awareness of food consumption, and ironic rebound as probable mediators. Results suggest that cognitive load may disinhibit consumption by preventing restrained eaters from monitoring the dietary consequences of their eating behavior. Implications for theories of self-regulation are discussed. PMID- 10794379 TI - Social comparisons of income in one's community: evidence from national surveys of income and happiness. AB - Two studies provide evidence for social comparison effects of income on subjective well-being (SWB). The 1st study of 7,023 persons from nationally representative samples in the United States shows that the range and skew of the income distribution in a community affects a person's happiness, as predicted by range-frequency theory. The 2nd study of 8 nations over a period of 25 years shows that decreasing the skew (inequality) of the income distribution in a country increases average national SWB. Both studies strongly support social comparison effects of income within a community, and both results are predicted by range-frequency theory. These studies are the first to successfully extend earlier results of R. H. Smith, E. Diener, and D. H. Wedell (1989) from the laboratory into naturalistic situations. The magnitude of the social comparison effects is smaller than the main effect of income, which implies that nations can avoid creating a "hedonic treadmill." PMID- 10794380 TI - Video games and aggressive thoughts, feelings, and behavior in the laboratory and in life. AB - Two studies examined violent video game effects on aggression-related variables. Study 1 found that real-life violent video game play was positively related to aggressive behavior and delinquency. The relation was stronger for individuals who are characteristically aggressive and for men. Academic achievement was negatively related to overall amount of time spent playing video games. In Study 2, laboratory exposure to a graphically violent video game increased aggressive thoughts and behavior. In both studies, men had a more hostile view of the world than did women. The results from both studies are consistent with the General Affective Aggression Model, which predicts that exposure to violent video games will increase aggressive behavior in both the short term (e.g., laboratory aggression) and the long term (e.g., delinquency). PMID- 10794381 TI - The relational-interdependent self-construal and relationships. AB - Three studies describe the development and validation of a measure of the relational-interdependent self-construal, which is defined as the tendency to think of oneself in terms of relationships with close others. Study 1 reports the development, psychometric properties, and tests of validity of this new measure. Individuals who scored high on the Relational-Interdependent Self-Construal (RISC) Scale characterized their important relationships as closer and more committed than did individuals who scored low on this measure (Study 1) and were more likely to take into account the needs and wishes of others when making decisions (Study 2). In Study 3, using a dyadic interaction paradigm with previously unacquainted participants, the partners of persons who scored high on the RISC scale viewed them as open and responsive to their needs and concerns; these perceptions were related to positive evaluations of the relationship. PMID- 10794382 TI - Vulnerable processes of nervous system development: a review of markers and methods. AB - The susceptibility of the developing nervous system to damage following exposure to environmental contaminants has been well recognized. More recently, from a regulatory perspective, an increased emphasis has been placed on the vulnerability of the developing nervous system to damage following pesticide exposure. The publication of the National Academy of Sciences (NAS) report on Pesticides in the Diets of Infants and Children (1995) and the passage of the Food Quality Protection Act (FQPA) and Safe Drinking Water Act (SDWA) amendments have significantly escalated the scientific debate regarding age-related susceptibility. Key concerns raised in the NAS report include the qualitative and quantitative differences that distinguish the developing nervous system from that of the adult. It was suggested that neurotoxicity testing on adult animals alone may not be predictive of these differences in susceptibility. The age-related susceptibility of the nervous system is compounded by the protracted period of time over which this complex organ system develops. This temporal vulnerability spans the embryonic, fetal, infant, and adolescent periods. Normal development of the nervous system requires the concomitant and coordinated ontogeny of proliferation, migration, differentiation, synaptogenesis, gliogenesis, myelination and apoptosis to occur in a temporally- and regionally-dependent manner. Perturbations of these processes during development can result in long term irreversible consequences that affect the structure and function of the nervous system and could account for qualitative differences in age-related susceptibility of the developing nervous system as compared to the adult nervous system. A discussion of developmental milestones and the relevance of transient effects on developmental endpoints are presented. Transient effects following developmental perturbations can be missed or dismissed depending on the experimental design or screening strategy employed. This subject is discussed in light of scientific uncertainties regarding perturbation-induced compensation in the developing nervous system. Thus, utilization of age-appropriate tests of these developmental processes may improve the detection and reduce uncertainty about the nature of adverse effects following developmental exposure to environmental neurotoxicants. PMID- 10794383 TI - Neonatal exposure to neurotoxic pesticides increases adult susceptibility: a review of current findings. AB - An environmental mischance commonly occuring in nature is the combination of neonatal exposure and later adult exposure to various toxic substances. During neonatal life, offspring can be affected by toxic agents either by transfer via mother's milk or by direct exposure. In many mammalian species the perinatal period is characterized by a rapid development of the brain--'the brain growth spurt' (BGS). We have observed that exposure to pesticides, such as DDT and bioallethrin, during the BGS in mice can potentiate susceptibility to bioallethrin or paraoxon in adult life. This combined neonatal and adult exposure caused spontaneous behavioural aberrations and changes in muscarinic cholinergic receptors and led to impairment of the faculties of learning and memory. Our studies indicate that neonatal exposure to pesticides--even in low doses--can potentiate and/or modify the reaction to adult exposure to xenobiotics, and thereby accelerate dysfunctional processes. PMID- 10794384 TI - Comparison of the role of esterases in the differential age-related sensitivity to chlorpyrifos and methamidophos. AB - More than 30 years ago, scientists recognized that, at a given dosage, the young rat was more sensitive than the adult to the toxicity of many organophosphorus, anticholinesterase pesticides. This observation went basically unexamined until recently. Renewed interest has emerged in scrutinizing this age-related sensitivity to pesticides, especially in light of the many new pesticides which are currently marketed. Our laboratory has been involved in comparing the age related sensitivity of young and adult rats to chlorpyrifos [Dursban, Lorsban; O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate] and methamidophos (Monitor; O,S-dimethyl phosphoamidothioate). Using chlorpyrifos, there is marked age-related sensitivity: direct oral dosing of the preweanling rat (postnatal day 17; PND17) with chlorpyrifos causes a toxic response (defined either behaviorally or biochemically) at a approximately 5-fold lower dosage than in adults (oral, maximum tolerated dose for the PND17 is 20 mg/kg versus 100 mg/kg for the adult). Other studies have indicated that the rat detoxifies chlorpyrifos and its oxon by binding to carboxylesterases and hydrolysis by A-esterases. The young rat is deficient in both these detoxification enzymes, which may explain the increased sensitivity of the young to chlorpyrifos toxicity. The age-related pattern for methamidophos is distinctly different: the oral, maximum tolerated dose is the same (8 mg/kg) whether the animal is 17 days old or an adult. We present data which indicate that methamidophos is not detoxified appreciably either in vivo or in vitro by A-esterases or carboxylesterases. Therefore, we submit the following hypothesis: organophosphorus pesticides, like chlorpyrifos, which are detoxified via A-esterases or carboxylesterases are more likely to exhibit age-related differences in sensitivity than pesticides which are not detoxified via these routes. PMID- 10794385 TI - A consideration of age-dependent differences in susceptibility to organophosphorus and pyrethroid insecticides. AB - Evidence that neonates are more sensitive than adults to organophosphorus (OP) and pyrethroid insecticides is largely based on studies that compare toxicity at acute lethal doses. Under such circumstances, the greater susceptibility of the neonate appears to be due to limited metabolic capacity rather than an inherent difference in the sensitivity of target sites. For purposes of risk assessment with food-use pesticides, the more relevant issue is whether the neonate is more sensitive than the adult to lower levels of exposure, approximating levels used to establish acceptable residue limits (tolerances) on various food products. If infants and children are not more sensitive to environmentally-relevant levels of exposure, then the existing tolerances for dietary exposure will provide adequate protection. If, on the other hand, they may be more sensitive, then additional studies with young animals or an additional uncertainty factor may be needed for added protection. This paper examines two sets of studies that address this issue. The first involves multi-generation reproduction studies with rats that were treated with OP insecticides (coumaphos, fenamiphos, tribufos, trichlorfon, or oxydemeton-methyl) through the diet and examined for effects, including cholinesterase (ChE) inhibition. The second involves rats that were treated by gavage with an acute dose of a pyrethroid (cismethrin, permethrin, deltamethrin or cypermethrin) to establish relative sensitivity to either a lethal dose or to a low, behaviorally-active dose. The results with the OP insecticides support ChE inhibition as the most sensitive measure of exposure and the critical effect (i.e., the lowest NOEL) for each study was based on ChE inhibition in the adult. The magnitude of ChE inhibition in pups (measured on postnatal day (PND) 4 and 21) was consistently less than for adults at a given dietary level. For the representative Type I pyrethroids, there was no evidence that pups are more sensitive than adults at any dose level. For both Type II pyrethroids, young rats were considerably more sensitive than adults to a lethal dose but not to lower doses. Levels of deltamethrin in whole-brain tissue support kinetics as the basis for the greater sensitivity of young rats to a lethal dose, with the immature systems involved with detoxification being overwhelmed at such high dose levels. These findings indicate that young animals are not more sensitive than adults to lower doses of OP or pyrethroid insecticides. This outcome supports the conclusion that infants and children are protected by existing tolerances, without the need for an additional uncertainty factor. PMID- 10794386 TI - Vulnerability to pesticide neurotoxicity is a lifetime issue. AB - Early development is not the only life stage during which which we see intensified responses to the adverse effects of chemicals. Vulnerability to toxic processes rises again late in life, and in many ways recapitulates the imperfect defenses deployed by the immature organism. One feature common to both early and late phases is a reduced capacity to compensate for impairment. In the first case, the functional mechanisms have yet to evolve. In the second, they have passed into what might be called a post-mature decline. Traced across the life cycle, this progression might be depicted as an inverted U. The developing brain, however, is equipped with immense plastic potential; the aging brain has lost much of its plasticity. The altered function of the aging brain, however, is not simply an outcome of how long the organism has lived. "Aging" is not a mechanistic explanation. Events occurring during life must account for the changes. Older brains are already high-maintenance properties, so that exposure to substances with neurotoxic properties, such as pesticides, may accelerate the process, or exploit its dwindling capacities to resist their effects. From this vantage point, toxicants can act in three ways to depress function during advanced age: they may interfere with brain development, leaving a legacy of diminished redundancy not apparent until it is further compromised during aging; they may hasten the progressive erosion of function observed with certain abilities; they may exert greater effects in the aging brain because the aging nervous system has already undergone a reduction in its ability to withstand toxic challenges. PMID- 10794387 TI - Organophosphate pesticides, cholinergic function and cognitive performance in advanced age. AB - The present communication will address the question of whether older individuals/animals are at greater risk to the cognitive impairment induced by exposure to organophosphates (OPs). There is considerable evidence for a decline in cholinergic indices (choline acetyltransferase, acetylcholinesterase, and muscarinic acetylcholine receptors) with aging. Whether these decreases lead to a greater or smaller response to OPs will depend on the relative rate of change in the specific indices. Some studies have shown that aged animals are more sensitive to cholinergic (muscarinic) agonists, even though the receptors are reduced. These findings suggest that aged individuals may be more sensitive to OPs. There is also much evidence documenting the decline in cognitive function in aged animals, a process which is accentuated in Alzheimer's disease and which has been attributed, in part, to the decline in cholinergic indices. Several studies have reported an improvement in cognitive functioning in aged animals/humans following treatment with acetylcholinesterase inhibitors. Thus, it is possible that the cognitive impairment, which occurs during aging, may be partially counteracted during treatment with certain cholinesterase inhibitors (both OPs and nonOPs). Whether the cognitive impairment reported by younger individuals some time after exposure to OPs would be greater in the elderly cannot be predicted on the basis of the available evidence. However, the findings in an animal model of dietary-induced cholinergic hypofunction developed by Roger Russell and colleagues may shed light on this question. PMID- 10794388 TI - The emerging field of ecogenetics. AB - Genetic differences in biotransformation enzymes and in target proteins can affect the individual susceptibility to drugs and environmental chemicals. The field of ecogenetics has emerged from the older area of pharmacogenetics, and investigates how genetic polymorphisms may represent risk factors for a number of diseases associated with exposure to toxic chemicals. Here, two polymorphisms, aldehyde dehydrogenase and delta-aminolevulinic acid dehydratase, are briefly discussed in relationship to alcohol and lead toxicity, respectively. The role of genetic polymorphisms in neurodegenerative diseases, such as Parkinson's disease, is also discussed. Furthermore, issues related to the functional significance of genetic polymorphisms, their interaction/combination, and ethical and societal considerations, are briefly addressed. PMID- 10794390 TI - Biotransformation enzyme polymorphism and pesticide susceptibility. AB - Numerous specific genetic polymorphisms (PM) in the multi-gene families of cytochromes P450 (CYPs) and glutathione S-transferases (GSTs) have been described in the human population in the past decade. For example, one or more PM have been identified in human CYP1A1, CYP1B1, CYP2C9, CYP2C18, CYP2D6, and CYP2E1. Recent studies using cDNA expressed human CYPs have suggested that CYP3A4 is the principal human CYP involved in the oxidation of parathion and probably other organo(thio)phosphate (OP) insecticides and thus PM in this CYP might influence susceptibility to OP. However, although large (> 10-fold) variability in CYP3A4 activity in human liver has been found, thus far no genetic basis for differences in activity or expression of CYP3A4 have been identified. Three GSTs are also polymorphic in the human population. Approximately 50% of the Caucasian population are homozygous for a gene deletion of the mu class GSTM1, and approximately 20% of Caucasians and over 60% of certain Asian populations are homozygous for a partial deletion of the theta-class GSTT1. Recently, several single nucleotide polymorphisms in human GSTP1 have also been described, and have altered activity toward several substrates. No studies have yet determined the relative activities of human GSTM1, T1 or PI towards methylparathion or other pesticides, and thus the potential significance of the common polymorphisms of these genes on pesticide susceptibility is unknown. Numerous studies have demonstrated that resistance of a variety of insects to several different insecticides, including DDT, has been attributed to the overexpression of theta class GSTs as well as certain CYPs. Thus, it remains possible that genetic PM in human GSTs and/or CYP enzymes could increase or decrease sensitivity to certain pesticides. Few epidemiological studies have examined whether any of the known CYP or GST PMs are associated with adverse outcomes in populations occupationally exposed to pesticides. PMID- 10794389 TI - Genetic and temporal determinants of pesticide sensitivity: role of paraoxonase (PON1). AB - Susceptibility to organophosphorus (OP) insecticides and nerve agents is strongly influenced by genetic and developmental factors. A number of organophosphorothioate insecticides are detoxified in part via a two-step pathway involving bioactivation of the parent compound by the cytochrome P450 systems, then hydrolysis of the resulting oxygenated metabolite (oxon) by serum and liver paraoxonases (PON1). Serum PON1 has been shown to be polymorphic in human populations. The Arg192 isoform (PON1R192) of this HDL-associated protein hydrolyzes paraoxon (POX) at a high rate, while the Gln192 isoform (PON1Q192) hydrolyzes paraoxon at a low rate. The effect of the polymorphism is reversed for the hydrolysis of diazoxon (DZO), soman and particularly sarin. Phenylacetate is hydrolyzed at approximately the same rate by both PON1 isoforms and chlorpyrifos oxon (CPO) slightly faster by the PON1R192 isoform. In addition to the effect of the amino acid substitution on rates of toxicant hydrolysis, two other factors influence these rates. The expression of PON1 is developmentally regulated. Newborns have very low levels of PON1. Adult levels in rats and mice are reached at 3 weeks of age and in humans, sometime after 6 months of age. In addition, among individuals of a given genotype, there is at least a 13-fold difference in expression of PON1 that is stable over time. Dose/response experiments with normal mice injected with purified PON1 and with PON1 knockout mice have clearly demonstrated that the observed differences of in vitro rates of hydrolysis are significant in determining differential sensitivities to specific insecticides processed through the P450/PON1 pathway. Injection of purified rabbit PON1 protects mice from cholinesterase inhibition by chlorpyrifos (CPS) and CPO. Knockout mice are much more sensitive to CPO and DZO than are their PON1+/+ littermates or wild-type mice. A number of recent reports have also indicated that the PON1R192 isoform may be a risk factor for cardiovascular disease. Studies with PON1 knockout mice are also consistent with a role of PON1 in preventing vascular disease. PMID- 10794391 TI - Pesticides and susceptible populations: people with butyrylcholinesterase genetic variants may be at risk. AB - Butyrylcholinesterase (BChE) scavenges low doses of organophosphorus (for example, paraoxon) and carbamate pesticides (for example, carbaryl) and in this way protects people from the toxic effects of these poisons. The protective role of BChE is demonstrated by the finding that pesticide applicators can have reduced BChE activity with no clinical signs of poisoning. The question has arisen whether people with genetic variants of BChE are less protected. Seventy six percent of the population is homozygous for wild-type BChE, while 24% carry at least one genetic variant allele. Most genetic variants of BChE have reduced activity. The clinically most important variant is atypical (D70G) BChE because people with this variant have 2 hours of apnea after receiving a dose of succinylcholine that is intended to paralyze muscles for 3-5 minutes. In test tube experiments the atypical variant reacts more slowly with all positively charged compounds (for example physostigmine, echothiophate). This leaves more toxin available for reaction with acetylcholinesterase in nerve synapses and predicts that people with atypical BChE will be less protected. Variants with low activity, such as silent BChE, are predicted to be at increased risk from organophosphorus pesticides based on experiments in monkeys and rodents where injection of purified BChE protected animals from the toxic effects of nerve agents. More studies are needed to strengthen the hypothesis that people with genetic variants of BChE are at higher risk of intoxication from pesticides. PMID- 10794392 TI - Differential sensitivity of sodium channel isoforms and sequence variants to pyrethroid insecticides. AB - Pyrethroids are commonly regarded as safe insecticides. However, some widely used pyrethroids, particularly single neurotoxic isomers of potent Type II compounds, have acute oral toxicities comparable to many organophosphorus insecticides. The majority of studies of the action of pyrethroids on voltage-sensitive sodium channels, the principal target sites for these compounds, have not considered differences in sodium channel structure as determinants of sensitivity. In mammals, voltage-sensitive sodium channels are encoded by a multi-gene family and exhibit both anatomical and developmental regulation of expression. Studies in this laboratory using cloned rat sodium channel isoforms expressed in Xenopus oocytes have documented profound differences in pyrethroid sensitivity between isoforms. Although the role of sodium channel gene mutations in altering pyethroid sensitivity has not been addressed in the case of the mammalian sodium channel gene family, the potential significance of allelic variation is illustrated in studies of point mutations in a sodium channel gene of the house fly that confer resistance to the lethal actions of pyrethroids and modify the sensitivity of house fly sodium channels expressed in Xenopus oocytes to these compounds. It is of particular interest that some of these resistance-associated mutations in the fly sodium channel occur at amino acid residues that are also the sites of mutations in human skeletal muscle sodium channels that are associated with inherited paralytic disorders. These findings document the pharmacological significance of structural differences between sodium channel isoforms and between genetic variants of an individual isoform as determinants of pyrethroid sensitivity. PMID- 10794393 TI - In vitro and in vivo effects of chlorpyrifos on glutathione peroxidase and catalase in developing rat brain. AB - Preliminary findings of a study on the role of oxidative stress in the developmental neurotoxicity of chlorpyrifos (CPF) indicates that in vitro exposure to 1-100 microM CPF or 1-100 nM CPF-oxon had no effect on the activity of glutathione peroxidase (GSHpx) in brain homogenates from postnatal day (PN) 21 rats, or on the activity of purified GSHpx. A single high-dose acute injection of 45 mg/kg CPF to PN19 rats also did not significantly alter GSHpx activity at PN21, in spite of extensive (72%) brain acetylcholinesterase (AChE) inhibition. However, catalase activity was significantly reduced by 28%. PN21 pups exposed maternally to a lower effective dose of CPF throughout development (dams injected with 50 mg/kg every 3 days) also had normal GSHpx activity, but a 30% increase in H2O2-independent NADPH consumption. Brain catalase activity in these rats was significantly increased by 24%. These preliminary data suggest that specific GSHpx activity is not altered by in vitro or in vivo exposures to CPF-oxon or CPF, but catalase and an unknown H2O2-independent NADPH-consuming factor were affected differentially depending on the type and timing of exposure. PMID- 10794394 TI - Detection of 2,4-dichlorophenoxyacetic acid (2,4-D) residues in neonates breast fed by 2,4-D exposed dams. AB - Knowing early nutritional status have been shown to be an important factor in determining the activity level of rats later in life, we studied offspring of dams which had received 50, 70 or 700 mg/kg of 2,4-Dichlorophenoxyacetic acid (2,4-D) during nursing. Neonatal tissues and the stomach content (milk) were examined up to 16 post natal days to detect body and organs weight alterations and 2,4-D residues after 2,4-D maternal dosing every-other-day, from post natal day 1. We detected 2,4-D residues in stomach content, blood, brain and kidney of 4-day-old neonates breast-fed by 2,4-D exposed mothers and onward. 2,4-D residues were dose- and exposure-time-dependent. The highest dose impaired body growth, as well as low tissue weights and diminished stomach contents. Levels of 2,4-D residues in stomach content, blood, kidney and brain of post natal rats (age PD 4 PD 16) fed through lactation from dams treated with 2,4-D demonstrated that 2,4-D was transferred to the neonates and the diminished body and tissues weight during this developmental period could be due to a diminished milk intake or/and to the direct 2,4-D toxic effect. Besides, when the herbicide treatment (100 mg 2,4 D/kg) was withdrawn from the dams, 2,4-D residues remained in the stomach content of neonates for at least one week. This fact indicated that dams continued depurating the herbicide through their milk. PMID- 10794395 TI - 2,4-Dichlorophenoxyacetic acid in developing rats alters behaviour, myelination and regions brain gangliosides pattern. AB - Neonate rats were exposed to 100 mg/kg from the PND 7 to PND 25 or at 70 mg/kg from PND 12 to PND 25. Treated and control pups were subjected to several behavioral tests (righting reflex, negative geotaxy, forelimbs support and open field) during the period of treatment. At PND 25 the regional effects of 2,4-D on gangliosides composition and myelin deposition were determined. The results indicate that the first design of exposure to 2,4-D produces significant diminutions in body and brain weight from PND 21. Furthermore, these pups showed decrease in GM1 level, diminution in myelin deposition and alterations in all behavioral tests. On the other hand, when treatment was not too severe (minor dose and shorter period of treatment), pups showed alterations in forelimb support and in open field tests without body or brain weight modifications. They also presented diminutions in GM1, mayoritary ganglioside of myelin, and in myelin deposition. These results suggest that in this latter 2,4-D exposure design, undernutrition could not be involved. PMID- 10794396 TI - Brain hypoplasia caused by exposure to trichlorfon and dichlorvos during development can be ascribed to DNA alkylation damage and inhibition of DNA alkyltransferase repair. AB - Treatment of pregnant guinea pigs with trichlorfon causes cerebellar hypoplasia in offspring. The most sensitive period for treatment is days 42-47 of gestation, which coincides with the rapid brain growth spurt and with the development of cerebellar granule cells. When rat granule cells were exposed in vitro to trichlorfon and dichlorvos for 24 hours they died, whereas trichloroethanol had no effect. When the cells were exposed to trichlorfon and dichlorvos for 3 hours, only dichlorvos was lethal indicating that the metabolite dichlorvos was more potent than trichlorfon itself. Cultured cerebellar granule cells were also found to be quite sensitive to other DNA-alkylating agents such as methylazoxymethanol and methylmethane sulphonate and to O6-benzylguanine; a potent and specific inhibitor of the DNA alkyltransferase involved in the repair of DNA alkylation damage. The organophosphorous compounds were also found to cause inhibition of the alkyltransferase and the lethal effects of the tested compounds on granule cell culture correlated well with the potency of inhibition. In a bacterial test system for monitoring alkylation effects on the DNA, dichlorvos was demonstrated to have a strong DNA alkylation effect. These results suggest that alkylation of DNA and inhibition of its repair can contribute to the brain hypoplasia observed after exposure to trichlorfon and dichlorvos during brain development. PMID- 10794397 TI - Interactions between pesticides and glia: an unexplored experimental field. AB - It is now well established that the role of astrocytes extends well beyond passive cytoskeletal structural support to neurons. In fact, astrocytes and neurons establish a highly dynamic reciprocal relationship that influences the growth, morphology, behavior, and central nervous system (CNS) repair. It is also well established that acute exposure to the organophosphorous insecticides leads to inhibition of neuronal acetylcholinesterase activity, an enzyme responsible for the inactivation of the neurotransmitter acetylcholine. Although astrocytes are known to express transport systems for choline, as well as acetylcholinesterase activity, little information is available on the potential interactions between the anticholinesterase class organophosphorous insecticides and these cells. This review will focus on astrocytic cholinergic receptors, choline uptake and metabolism, and address the potential importance of astrocytes in organophosphorous insecticide mediated neurotoxicity. PMID- 10794398 TI - The food quality protection act: a public health perspective. AB - The Food Quality Protection Act (FQPA) includes provisions to consider the combined risks of chemicals that have similar toxicological effects from all sources of exposure. The organophosphate (OP) insecticides are drawing a great deal of attention under this provision. Significant challenges for the regulatory activities at the EPA may develop. Environmentalists will apply political pressure to ban most of these products. Farmers may fear they will be unable to control pests and may exercise their own political muscle. The regulatory decisions in the dilemma may prove to be complex and difficult. Three areas of concern about potential adverse impacts of FQPA on public health are discussed; these include: diseases caused or carried by pests, increased natural toxins because of poor pest control, and increases in food costs because of regulatory action. It is important that the EPA consider not only the risks arising from the use of chemicals, but also the risks of not having those same chemicals. PMID- 10794399 TI - Neurotoxicology risk assessment guidelines: developmental neurotoxicology. AB - EPA's Neurotoxicity Risk Assessment Guidelines were recently published in final form in the Federal Register (1998). This document was developed over a period of nearly ten years and is intended to establish operating principles used in the evaluation of data for neurotoxicity risk assessment. The guidelines contain a number of assumptions and definitions of key concepts, as well as guidance as to the evaluation of various behavioral and structural changes produced by chemical exposure in humans and animals. With regard to developmental neurotoxicity, risk assessors should be aware that chemical-induced neurotoxicity in adults may not always be a good predictor of developmental neurotoxicity. Adverse effects on the developing nervous system can occur prior to conception up to the time of sexual maturity, depend on the time of exposure relative to a critical state of nervous system development, can be seen at any time during the lifespan of the organism, may lead to delayed onset or latent effects, and may elicit compensatory mechanisms that obscure underlying neurotoxicity. Adverse effects include persistent alterations in function or structure of the nervous system or a change in the time or appearance of any endpoint. Relative to neurotoxicity in adult animals, there are several special concerns in hazard characterization of developmental studies, including maternal toxicity, the use of the litter as the statistical unit, and time of exposure relative to the ontogeny of various structural or functional endpoints. Dose-response evaluation of data from developmental studies is similar to that for adults, although a safety factor of 10 may be applied to protect children's health. The guidelines also note that exposure patterns of children differ from those of adults resulting in a greater intake of chemicals on a per body weight basis. The guidelines note several research needs, including more information on mechanisms of developmental neurotoxicity, mechanistically based dose-response models, impact of early exposure to chemicals on late-onset disease, studies on threshold, and experiments on potential interactions between chemicals in mixtures. PMID- 10794400 TI - Do pesticides reduce our total exposure to food borne toxicants? AB - In addition to the long history of disease and death from natural toxicants in plants, there is a growing literature about the abundant levels and varieties of natural toxicants in our foods. These natural chemicals cause mutagenicity, carcinogenicity, teratogenicity, neurotoxicity and visceral organ toxicity in routine laboratory tests. While many of these chemicals occur at rather fixed levels in foods, there is evidence that a large number of natural toxicants increase or decrease with levels of plant stress. When plants are stressed from infection or predation, they characteristically respond with a rapid increase in defensive chemicals. Because increases in natural toxicants are stress mediated, the prevention of infection and predation can reduce the levels of the natural toxicants in our food supply. In addition, fungal infections induce a chemical response from the plant, and also excrete their own array of toxins, sometimes at high levels. Crop protection chemicals are used to reduce plant stress, but there is no program today to balance the risk from crop protection chemicals (which are extensively tested and heavily regulated) against the possible benefit of decreased risk from natural plant and fungal toxicants (which are only sporadically tested and are largely unregulated). Because crop protection chemicals are logically linked to inhibition of the expression of natural toxicants, there may be considerable food safety benefits from an approach to pesticide risk management that evaluates the risk versus risk equation. On the other hand, inattention to the issue of suppression of natural toxicants may lead to an unintended increase in our exposure to this wide array of naturally occurring chemicals. PMID- 10794401 TI - Chlorpyrifos: probabilistic assessment of exposure and risk. AB - FQPA requires determination of the combined potential risk from both dietary (food and drinking water) and non-dietary (residential) routes of exposures to pesticides. Critical to the implementation and ultimately to the impact of FQPA are the validity of the risk assessments. Fundamental to the practice of sound science in risk assessment is accuracy in representing real-world exposures and the resulting risk. Too often, simplistic and overly conservative approaches and exposure data are employed. The resulting assessments are often termed "worst case" and the inherent inaccuracies justified as additional safety factors. A scientific, tiered approach can be employed to improve the accuracy of risk assessments and provide more realistic estimates of risk. A case study using chlorpyrifos, a widely-studied insecticide, will be presented to show the levels of refinement which can be obtained through the use of more relevant exposure data, recognition of the patterns of uses and exposures, and higher-tier probabilistic methodologies. Results show a 7- to 26-fold decrease in estimated acute dietary risk and a 30- to 80-fold decrease in estimated aggregate risk using these refinements. These decreases proved to be critical, since the change in results also change the conclusions about risk and indicate the levels are within acceptable limits. PMID- 10794402 TI - Our changing perspectives on benefits and risks of pesticides: a historical overview. AB - The introduction of chemical pesticides following WW II ushered in the era of the "quick fix" for any aqricultural, forestry and human health problems. Scenarios of use, misuse, abuse and environmental contamination can be presented for any class of pesticide, culminating in dependence on these chemicals for increased production of food and fibre and improved health. With time, sophisticated agents having unique, target-specific mechanisms of action evolved but at increased cost(s) to crop production. Equatorial countries, rapidly becoming "breadbaskets" of the world, are particularly dependent on pesticides as they strive to increase production of nontraditional export products (NTEPS), valuable cash crops in demand in countries having more temperate climates. Developing nations have neither the legislation and regulations necessary to control pesticides nor trained personnel to inspect and monitor use, to analyze residues in produce or to initiate training programs. Their transition from agrarian to industrialized societies has meant that smaller, less well educated populations must shoulder the responsibility of increased traditional food production for consumption by urban populations as well as that of NTEPS. Unfortunately, to attain these goals, many older, more toxic, environmentally persistent and cheap pesticides, long banned in developed countries, are used extensively, creating serious local and global contamination and health problems. PMID- 10794403 TI - Pesticide neurotoxicity in Europe: real risks and perceived risks. AB - The same classes of pesticides are used all over the world, but conditions of use vary widely, and public perceptions of risk vary more widely still. Within Western Europe pesticide residues in commercially traded foodstuffs are subject to international standards and are closely monitored. Hence risks to consumers from such foods are negligible. The major hazards are poisoning associated with high acute/chronic operator exposures due to occasional pesticide misuse. In addition pesticides provide a convenient means of attempting suicide in agricultural areas. In contrast, public perception of risks from pesticides centres on low level exposures, and is heightened by several factors. These are: poisonings associated with pesticide misuse; the indirect nature of the benefit to the consumer (cf. medicines or public health uses); and commercially motivated marketing of pesticide-free produce. In press reports pyrethroid insecticides have been linked to "Multiple Chemical Sensitivity" in Germany, and organophosphates to "Chronic Fatigue Syndrome" in the UK. A number of pressure groups are actively campaigning to ban all uses of organophosphorus pesticides. Unfortunately evaluation of the real risks of pesticide exposure is rendered less certain by the lack of any very useful retrospective exposure measures with which biological effects of uncertain aetiology might be correlated. This means that although we can be sure that pesticides pose no gross threat to health in the general population, subtle effects on more highly exposed sub-populations are, as yet, more difficult to rule out. PMID- 10794404 TI - Organophosphates and their impact on the global environment. AB - Serious intoxications and incidences due to misuse of organophosphorus insecticides (OP) have been reported for over three decades. In this meeting I am talking about the following three topics. (1) Epidemiological studies on the use of OP in Japan, Taiwan and Thailand. The National Research Institute of Police Science (NRIPS) in Japan has published the annual report of the epidemiological studies on the criminal and suicide events. The numbers of the incidence of pesticides are approximately 22%-30% of the total incidences, during 1991 and 1996. The incidence of pesticides has been gradually reduced for past six years. This seems to be influenced by the decrease of the incidences of paraquat having extremely high toxicity. According to the epidemiological data in the National Poison Center in Taiwan, unlike the prevalence of drug poisoning exposures in most western countries, pesticides poisoning exposures are numerous in Taiwan. The number of pesticide intoxication cases is 6,872 out of 23,436 of total case numbers of various intoxications during the 1985 and 1997. The fatality rate is much higher in Taiwan than in most western countries, with 5.65% of all poisoning exposures resulted in death. Paraquot is the leading cause of death in Taiwan. The ratio of death to total OP intoxication cases is 11.5% including accidental exposure and suicide. According to the report of the Division of Epidemiology, Ministry of Public Health, Thailand during the 1990-1995, the overall morbidity due to poisoning was 30 per 100,000 population with 300 deaths per year. The morbidity rate of reported pesticide poisoning in the whole country was 10 per 100,000 population. (2) Sarin victims in Tokyo. On March 20, 1995, a terrorist attack using sarin (isopropylmethylphosphonofluoridate) occurred on the Tokyo subway. Many people inhaled the sarin gas and collapsed in the train. Eventually, 12 people died and over 5,000 were injured. The results of the forensic studies showed that methylphosphonic acid, the hydrolysis product of sarin, bound to AChE in the cerebellums of the victims was separated and identified using GCMS. (3) A sensitive and rapid biomarker of OP-poisoning. Beta-glucuronidase (beta-G) is loosely bound to egasyn which is one of carboxylesterase isozymes in the liver microsomes. The oxon formed from OP in the liver microsomes is covalently bound to egasyn and causes the cleavage of the egasyn-glucuronidase complex, leading to the release of beta-G into the blood. In fact, plasma beta-G activities is a much more sensitive and rapid biomarker than the blood cholinesterase inhibition. In conclusion, OPs cause serious toxicity in both acute and chronic exposures. The epidemiological data in Taiwan and Thailand show that paraquat intoxication cause the most serious irreversible damage in the patients. In 1995, terrorists attack using sarin which is one of the most toxic OP, on the Tokyo subway, and 12 people died. This is an example of the acute intoxication of OP and some people have been still in hospitalized. Recently, the present authors have reported that the plasma beta-G activity is a novel biomarker of OP intoxication. The plasma beta-G is rapidly and significantly increased after OP exposure, and this is a much more sensitive to OP intoxication than ChE inhibition. PMID- 10794405 TI - A helix-turn motif in the C-terminal domain of histone H1. AB - The structural study of peptides belonging to the terminal domains of histone H1 can be considered as a step toward the understanding of the function of H1 in chromatin. The conformational properties of the peptide Ac EPKRSVAFKKTKKEVKKVATPKK (CH-1), which belongs to the C-terminal domain of histone H1(o) (residues 99-121) and is adjacent to the central globular domain of the protein, were examined by means of 1H-NMR and circular dichroism. In aqueous solution, CH-1 behaved as a mainly unstructured peptide, although turn-like conformations in rapid equilibrium with the unfolded state could be present. Addition of trifluoroethanol resulted in a substantial increase of the helical content. The helical limits, as indicated by (i,i + 3) nuclear Overhauser effect (NOE) cross correlations and significant up-field conformational shifts of the C(alpha) protons, span from Pro100 to Val116, with Glu99 and Ala117 as N- and C caps. A structure calculation performed on the basis of distance constraints derived from NOE cross peaks in 90% trifluoroethanol confirmed the helical structure of this region. The helical region has a marked amphipathic character, due to the location of all positively charged residues on one face of the helix and all the hydrophobic residues on the opposite face. The peptide has a TPKK motif at the C-terminus, following the alpha-helical region. The observed NOE connectivities suggest that the TPKK sequence adopts a type (I) beta-turn conformation, a sigma-turn conformation or a combination of both, in fast equilibrium with unfolded states. Sequences of the kind (S/T)P(K/R)(K/R) have been proposed as DNA binding motifs. The CH-1 peptide, thus, combines a positively charged amphipathic helix and a turn as potential DNA-binding motifs. PMID- 10794406 TI - Elucidation of the solution structure of cardiotoxin analogue V from the Taiwan cobra (Naja naja atra)--identification of structural features important for the lethal action of snake venom cardiotoxins. AB - The aim of the present study is to understand the structural features responsible for the lethal activity of snake venom cardiotoxins. Comparison of the lethal potency of the five cardiotoxin isoforms isolated from the venom of Taiwan cobra (Naja naja atra) reveals that the lethal potency of CTX I and CTX V are about twice of that exhibited by CTX II, CTX III, and CTX IV. In the present study, the solution structure of CTX V has been determined at high resolution using multidimensional proton NMR spectroscopy and dynamical simulated annealing techniques. Comparison of the high resolution solution structures of CTX V with that of CTX IV reveals that the secondary structural elements in both the toxin isoforms consist of a triple and double-stranded antiparallel beta-sheet domains. Critical examination of the three-dimensional structure of CTX V shows that the residues at the tip of Loop III form a distinct "finger-shaped" projection comprising of nonpolar residues. The occurrence of the nonpolar "finger-shaped" projection leads to the formation of a prominent cleft between the residues located at the tip of Loops II and III. Interestingly, the occurrence of a backbone hydrogen bonding (Val27CO to Leu48NH) in CTX IV is found to distort the "finger-shaped" projection and consequently diminish the cleft formation at the tip of Loops II and III. Comparison of the solution structures and lethal potencies of other cardiotoxin isoforms isolated from the Taiwan cobra (Naja naja atra) venom shows that a strong correlation exists between the lethal potency and occurrence of the nonpolar "finger-shaped" projection at the tip of Loop III. Critical analysis of the structures of the various CTX isoforms from the Taiwan cobra suggest that the degree of exposure of the cationic charge (to the solvent) contributed by the invariant lysine residue at position 44 on the convex side of the CTX molecules could be another crucial factor governing their lethal potency. PMID- 10794407 TI - Mutational analysis of the major coat protein of M13 identifies residues that control protein display. AB - We have reported variants of the M13 bacteriophage major coat protein (P8) that enable high copy display of monomeric and oligomeric proteins, such as human growth hormone and steptavidin, on the surface of phage particles (Sidhu SS, Weiss GA, Wells JA. 2000. High copy display of large proteins on phage for functional selections. J Mol Biol 296:487-495). Here, we explore how an optimized P8 variant (opti-P8) could evolve the ability to efficiently display a protein fused to its N-terminus. Reversion of individual opti-P8 residues back to the wild-type P8 residue identifies a limited set of hydrophobic residues responsible for the high copy protein display. These hydrophobic amino acids bracket a conserved hydrophobic face on the P8 alpha helix thought to be in contact with the phage coat. Mutations additively combine to promote high copy protein display, which was further enhanced by optimization of the linker between the phage coat and the fusion protein. These data are consistent with a model in which protein display-enhancing mutations allow for better packing of the fusion protein into the phage coat. The high tolerance for phage coat protein mutations observed here suggests that filamentous phage coat proteins could readily evolve new capabilities. PMID- 10794408 TI - A method to predict residues conferring functional differences between related proteins: application to MAP kinase pathways. AB - Physicochemical properties are potentially useful in predicting functional differences between aligned protein subfamilies. We present a method that considers physicochemical properties from ancestral sequences predicted to have given rise to the subfamilies of interest by gene duplication. Comparison between two map kinases subfamilies, p38 and ERK, revealed a region that had an excess of change in properties after gene duplication followed by conservation within the two subfamilies. This region corresponded to that experimentally defined as important for substrate and pathway specificity. The derived scores for the region of interest were found to differ significantly in their distribution compared to the rest of the protein when the Kolmogorov-Smirnov test was applied (p = 0.005). Thus, the incorporation of ancestral physicochemical properties is useful in predicting functional differences between protein subfamilies. In addition, the method was applied to the MKK and MAPK components of the p38 and JNK pathways. These proteins showed a similar pattern in their evolution and regions predicted to confer functional differences are discussed. PMID- 10794409 TI - Backbone dynamics measurements on leukemia inhibitory factor, a rigid four helical bundle cytokine. AB - The backbone dynamics of the four-helical bundle cytokine leukemia inhibitory factor (LIF) have been investigated using 15N NMR relaxation and amide proton exchange measurements on a murine-human chimera, MH35-LIF. For rapid backbone motions (on a time scale of 10 ps to 100 ns), as probed by 15N relaxation measurements, the dynamics parameters were calculated using the model-free formalism incorporating the model selection approach. The principal components of the inertia tensor of MH35-LIF, as calculated from its NMR structure, were 1:0.98:0.38. The global rotational motion of the molecule was, therefore, assumed to be axially symmetric in the analysis of its relaxation data. This yielded a diffusion anisotropy D(parallel)/D(perpendicular) of 1.31 and an effective correlation time (4D(perpendicular) + 2D(parallel))(-1) of 8.9 ns. The average values of the order parameters (S2) for the four helices, the long interhelical loops, and the N-terminus were 0.91, 0.84, and 0.65, respectively, indicating that LIF is fairly rigid in solution, except at the N-terminus. The S2 values for the long interhelical loops of MH35-LIF were higher than those of their counterparts in short-chain members of the four-helical bundle cytokine family. Residues involved in LIF receptor binding showed no consistent pattern of backbone mobilities, with S2 values ranging from 0.71 to 0.95, but residues contributing to receptor binding site III had relatively lower S2 values, implying higher amplitude motions than for the backbone of sites I and II. In the relatively slow motion regime, backbone amide exchange measurements showed that a number of amides from the helical bundle exchanged extremely slowly, persisting for several months in 2H2O at 37 degrees C. Evidence for local unfolding was considered, and correlations among various structure-related parameters and the backbone amide exchange rates were examined. Both sets of data concur in showing that LIF is one of the most rigid four-helical bundle cytokines. PMID- 10794410 TI - Oxygen binding by alpha(Fe2+)2beta(Ni2+)2 hemoglobin crystals. AB - Oxygen binding by hemoglobin fixed in the T state either by crystallization or by encapsulation in silica gels is apparently noncooperative. However, cooperativity might be masked by different oxygen affinities of alpha and beta subunits. Metal hybrid hemoglobins, where the noniron metal does not bind oxygen, provide the opportunity to determine the oxygen affinities of alpha and beta hemes separately. Previous studies have characterized the oxygen binding by alpha(Ni2+)2beta(Fe2+)2 crystals. Here, we have determined the three-dimensional (3D) structure and oxygen binding of alpha(Fe2+)2beta(Ni2+)2 crystals grown from polyethylene glycol solutions. Polarized absorption spectra were recorded at different oxygen pressures with light polarized parallel either to the b or c crystal axis by single crystal microspectrophotometry. The oxygen pressures at 50% saturation (p50s) are 95 +/- 3 and 87 +/- 4 Torr along the b and c crystal axes, respectively, and the corresponding Hill coefficients are 0.96 +/- 0.06 and 0.90 +/- 0.03. Analysis of the binding curves, taking into account the different projections of the alpha hemes along the optical directions, indicates that the oxygen affinity of alpha1 hemes is 1.3-fold lower than alpha2 hemes. Inspection of the 3D structure suggests that this inequivalence may arise from packing interactions of the Hb tetramer within the monoclinic crystal lattice. A similar inequivalence was found for the beta subunits of alpha(Ni2+)2beta(Fe2+)2 crystals. The average oxygen affinity of the alpha subunits (p50 = 91 Torr) is about 1.2-fold higher than the beta subunits (p50 = 110 Torr). In the absence of cooperativity, this heterogeneity yields an oxygen binding curve of Hb A with a Hill coefficient of 0.999. Since the binding curves of Hb A crystals exhibit a Hill coefficient very close to unity, these findings indicate that oxygen binding by T-state hemoglobin is noncooperative, in keeping with the Monod, Wyman, and Changeux model. PMID- 10794411 TI - 15N and 1H NMR study of histidine containing protein (HPr) from Staphylococcus carnosus at high pressure. AB - The pressure-induced changes in 15N enriched HPr from Staphylococcus carnosus were investigated by two-dimensional (2D) heteronuclear NMR spectroscopy at pressures ranging from atmospheric pressure up to 200 MPa. The NMR experiments allowed the simultaneous observation of the backbone and side-chain amide protons and nitrogens. Most of the resonances shift downfield with increasing pressure indicating generalized pressure-induced conformational changes. The average pressure-induced shifts for amide protons and nitrogens are 0.285 ppm GPa(-1) at 278 K and 2.20 ppm GPa(-1), respectively. At 298 K the corresponding values are 0.275 and 2.41 ppm GPa(-1). Proton and nitrogen pressure coefficients show a significant but rather small correlation (0.31) if determined for all amide resonances. When restricting the analysis to amide groups in the beta-pleated sheet, the correlation between these coefficients is with 0.59 significantly higher. As already described for other proteins, the amide proton pressure coefficients are strongly correlated to the corresponding hydrogen bond distances, and thus are indicators for the pressure-induced changes of the hydrogen bond lengths. The nitrogen shift changes appear to sense other physical phenomena such as changes of the local backbone conformation as well. Interpretation of the pressure-induced shifts in terms of structural changes in the HPr protein suggests the following picture: the four-stranded beta-pleated sheet of HPr protein is the least compressible part of the structure showing only small pressure effects. The two long helices a and c show intermediary effects that could be explained by a higher compressibility and a concomitant bending of the helices. The largest pressure coefficients are found in the active center region around His15 and in the regulatory helix b which includes the phosphorylation site Ser46 for the HPr kinase. This suggests that this part of the structure occurs in a number of different structural states whose equilibrium populations are shifted by pressure. In contrast to the surrounding residues of the active center loop that show large pressure effects, Ile14 has a very small proton and nitrogen pressure coefficient. It could represent some kind of anchoring point of the active center loop that holds it in the right place in space, whereas other parts of the loop adapt themselves to changing external conditions. PMID- 10794413 TI - Slow conformational dynamics of an endonuclease persist in its complex with its natural protein inhibitor. AB - The bacterial toxin colicin E9 is secreted by producing Escherichia coli cells with its 9.5 kDa inhibitor protein Im9 bound tightly to its 14.5 kDa C-terminal DNase domain. Double- and triple-resonance NMR spectra of the isolated DNase domain uniformly labeled with 13C/15N bound to unlabeled Im9 contain more signals than expected for a single DNase conformer, consistent with the bound DNase being present in more than one form. The presence of chemical exchange cross peaks in 750 MHz 15N-1H-15N HSQC-NOESY-HSQC spectra for backbone NH groups of Asp20, Lys21, Trp22, Leu23, Lys69, and Asn70 showed that the bound DNase was in dynamic exchange. The rate of exchange from the major to the minor form was determined to be 1.1 +/- 0.2 s(-1) at 298 K. Previous NMR studies have shown that the free DNase interchanges between two conformers with a forward rate constant of 1.61 +/ 0.11 s(-1) at 288 K, and that the bound Im9 is fixed in one conformation. The NMR studies of the bound DNase show that Im9 binds similarly to both conformers of the DNase and that the buried Trp22 is involved in the dynamic process. For the free DNase, all NH groups within a 9 A radius of any point of the Trp22 ring exhibit heterogeneity suggesting that a rearrangement of the position of this side chain is connected with the conformational interchange. The possible functional significance of this feature of the DNase is discussed. PMID- 10794412 TI - Crystal structure of adenosine kinase from Toxoplasma gondii at 1.8 A resolution. AB - Human infection with Toxoplasma gondii is an important cause of morbidity and mortality. Protozoan parasites such as T. gondii are incapable of de novo purine biosynthesis and must acquire purines from their host, so the purine salvage pathway offers a number of potential targets for antiparasitic chemotherapy. In T. gondii tachyzoites, adenosine is the predominantly salvaged purine nucleoside, and thus adenosine kinase is a key enzyme in the purine salvage pathway of this parasite. The structure of T. gondii adenosine kinase was solved using molecular replacement and refined by simulated annealing at 1.8 A resolution to an R-factor of 0.214. The overall structure and the active site geometry are similar to human adenosine kinase, although there are significant differences. The T. gondii adenosine kinase has several unique features compared to the human sequence, including a five-residue deletion in one of the four linking segments between the two domains, which is probably responsible for a major change in the orientation of the two domains with respect to each other. These structural differences suggest the possibility of developing specific inhibitors of the parasitic enzyme. PMID- 10794414 TI - NMR solution structure of the theta subunit of DNA polymerase III from Escherichia coli. AB - The catalytic core of Escherichia coli DNA polymerase III contains three tightly associated subunits (alpha, epsilon, and theta). The theta subunit is the smallest, but the least understood of the three. As a first step in a program aimed at understanding its function, the structure of the theta subunit has been determined by triple-resonance multidimensional NMR spectroscopy. Although only a small protein, theta was difficult to assign fully because approximately one third of the protein is unstructured, and some sections of the remaining structured parts undergo intermediate intramolecular exchange. The secondary structure was deduced from the characteristic nuclear Overhauser effect patterns, the 3J(HN alpha) coupling constants and the consensus chemical shift index. The C terminal third of the protein, which has many charged and hydrophilic amino acid residues, has no well-defined secondary structure and exists in a highly dynamic state. The N-terminal two-thirds has three helical segments (Gln10-Asp19, Glu38 Glu43, and His47-Glu54), one short extended segment (Pro34-Ala37), and a long loop (Ala20-Glu29), of which part may undergo intermediate conformational exchange. Solution of the three-dimensional structure by NMR techniques revealed that the helices fold in such a way that the surface of theta is bipolar, with one face of the protein containing most of the acidic residues and the other face containing most of the long chain basic residues. Preliminary chemical shift mapping experiments with a domain of the epsilon subunit have identified a loop region (Ala20-Glu29) in theta as the site of association with epsilon. PMID- 10794415 TI - An enigmatic peptide ligation reaction: protease-catalyzed oligomerization of a native protein segment in neat aqueous solution. AB - We report an enigmatic peptide ligation reaction catalyzed by Glu-specific Staphylococcus aureus V8 protease that occurs in neat aqueous solution around neutral pH utilizing a totally unprotected peptide substrate containing free alpha-carboxyl and alpha-amino groups. V8 protease catalyzed a chain of ligation steps between pH 6 and 8 at 4 degrees C, producing a gamut of covalent oligomers (dimer through octamer or higher) of a native protein segment TAAAKFE (S39) derived from ribonuclease A (RNAse A). Size-exclusion chromatography suggested the absence of strong interaction between the reacting peptides. The circular dichroism spectra of monomer through pentamer showed length-dependent enhancement of secondary structure in the oligomers, suggesting that protease-catalyzed ligation of a monomer to an oligomer resulted in a product that was more structured than its precursor. The relative conformational stability of the oligomers was reflected in their ability to resist proteolysis, indicating that the oligomerization reaction was facilitated as a consequence of the "conformational trapping" of the product. The ligation reaction proceeded in two phases-slow formation and accumulation of the dimer followed by a fast phase of oligomerization, implying that the conformational trap encountered in the oligomerization reaction was a two-step process. The Gly substitution at any position of the TAAAKFE sequence was deleterious, suggesting that the first step of the conformational trap, namely the dimerization reaction, that proceeded very slowly even with the parent peptide, was quite sensitive to amino acid sequence. In contrast, the oligomerization reaction of an Ala analog, AAAAKFE, occurred in much the same way as S39, albeit with faster rate, suggesting that Ala substitution stabilized the overall conformational trapping process. The results suggest the viability of the product-directed "conformational trap" as a mechanism to achieve peptide ligation of totally unprotected peptide fragments in neat aqueous solution. Further, the study projects the presence of considerable innate synthetic potential in V8 protease, baring rich possibilities of protein engineering of this enzyme to generate a "V8 peptide ligase." PMID- 10794417 TI - Determination of alpha-helix N1 energies after addition of N1, N2, and N3 preferences to helix/coil theory. AB - Surveys of protein crystal structures have revealed that amino acids show unique structural preferences for the N1, N2, and N3 positions in the first turn of the alpha-helix. We have therefore extended helix-coil theory to include statistical weights for these locations. The helix content of a peptide in this model is a function of N-cap, C-cap, N1, N2, N3, C1, and helix interior (N4 to C2) preferences. The partition function for the system is calculated using a matrix incorporating the weights of the fourth residue in a hexamer of amino acids and is implemented using a FORTRAN program. We have applied the model to calculate the N1 preferences of Gln, Val, Ile, Ala, Met, Pro, Leu, Thr, Gly, Ser, and Asn, using our previous data on helix contents of peptides Ac-XAKAAAAKAAGY-CONH2. We find that Ala has the highest preference for the N1 position. Asn is the most unfavorable, destabilizing a helix at N1 by at least 1.4 kcal mol(-1) compared to Ala. The remaining amino acids all have similar preferences, 0.5 kcal mol(-1) less than Ala. Gln, Asn, and Ser, therefore, do not stabilize the helix when at N1. PMID- 10794416 TI - Insect peptides with improved protease-resistance protect mice against bacterial infection. AB - At a time of the emergence of drug-resistant bacterial strains, the development of antimicrobial compounds with novel mechanisms of action is of considerable interest. Perhaps the most promising among these is a family of antibacterial peptides originally isolated from insects. These were shown to act in a stereospecific manner on an as-yet unidentified target bacterial protein. One of these peptides, drosocin, is inactive in vivo due to the rapid decomposition in mammalian sera. However, another family member, pyrrhocoricin, is significantly more stable, has increased in vitro efficacy against gram-negative bacterial strains, and if administered alone, as we show here, is devoid of in vitro or in vivo toxicity. At low doses, pyrrhocoricin protected mice against Escherichia coli infection, but at a higher dose augmented the infection of compromised animals. Analogs of pyrrhocoricin were, therefore, synthesized to further improve protease resistance and reduce toxicity. A linear derivative containing unnatural amino acids at both termini showed high potency and lack of toxicity in vivo and an expanded cyclic analog displayed broad activity spectrum in vitro. The bioactive conformation of native pyrrhocoricin was determined by nuclear magnetic resonance spectroscopy, and similar to drosocin, reverse turns were identified as pharmacologically important elements at the termini, bridged by an extended peptide domain. Knowledge of the primary and secondary structural requirements for in vivo activity of these peptides allows the design of novel antibacterial drug leads. PMID- 10794418 TI - Improving protein crystal quality by decoupling nucleation and growth in vapor diffusion. AB - A simple method for growing protein crystals in the metastable zone using the vapor diffusion technique is described. The coverslips holding the hanging drops are transferred, after being incubated for some time at conditions normally giving many small crystals, over reservoirs at concentrations that normally yield clear drops. Fewer, much larger and better diffracting crystals are obtained, compared with conventional crystallization at similar conditions. To our knowledge, this is the first report of a significant crystal improvement due to "backing off" from nucleation conditions, using the hanging drop method. A correlation of the transfer time with published results for vapor diffusion equilibration of poly(ethylene glycol) solutions is also presented. PMID- 10794419 TI - Specificity in substrate binding by protein folding catalysts: tyrosine and tryptophan residues are the recognition motifs for the binding of peptides to the pancreas-specific protein disulfide isomerase PDIp. AB - Using a cross-linking approach, we recently demonstrated that radiolabeled peptides or misfolded proteins specifically interact in vitro with two luminal proteins in crude extracts from pancreas microsomes. The proteins were the folding catalysts protein disulfide isomerase (PDI) and PDIp, a glycosylated, PDI related protein, expressed exclusively in the pancreas. In this study, we explore the specificity of these proteins in binding peptides and related ligands and show that tyrosine and tryptophan residues in peptides are the recognition motifs for their binding by PDIp. This peptide-binding specificity may reflect the selectivity of PDIp in binding regions of unfolded polypeptide during catalysis of protein folding. PMID- 10794420 TI - Analysis of knowledge-based protein-ligand potentials using a self-consistent method. AB - We propose a self-consistent approach to analyze knowledge-based atom-atom potentials used to calculate protein-ligand binding energies. Ligands complexed to actual protein structures were first built using the SMoG growth procedure (DeWitte & Shakhnovich, 1996) with a chosen input potential. These model protein ligand complexes were used to construct databases from which knowledge-based protein-ligand potentials were derived. We then tested several different modifications to such potentials and evaluated their performance on their ability to reconstruct the input potential using the statistical information available from a database composed of model complexes. Our data indicate that the most significant improvement resulted from properly accounting for the following key issues when estimating the reference state: (1) the presence of significant nonenergetic effects that influence the contact frequencies and (2) the presence of correlations in contact patterns due to chemical structure. The most successful procedure was applied to derive an atom-atom potential for real protein-ligand complexes. Despite the simplicity of the model (pairwise contact potential with a single interaction distance), the derived binding free energies showed a statistically significant correlation (approximately 0.65) with experimental binding scores for a diverse set of complexes. PMID- 10794421 TI - Novel disulfide engineering in human carbonic anhydrase II using the PAIRWISE side-chain geometry database. AB - An analysis of the pairwise side-chain packing geometries of cysteine residues observed in high-resolution protein crystal structures indicates that cysteine pairs have pronounced orientational preferences due to the geometric constraints of disulfide bond formation. A potential function was generated from these observations and used to evaluate models for novel disulfide bonds in human carbonic anhydrase II (HCAII). Three double-cysteine variants of HCAII were purified and the effective concentrations of their thiol groups were determined by titrations with glutathione and dithiothreitol. The effects of the cysteine mutations on the native state structure and stability were characterized by circular dichroism, enzymatic activity, sulfonamide binding, and guanidine hydrochloride titration. These analyses indicate that the PAIRWISE potential is a good predictor of the strength of the disulfide bond itself, but the overall structural and thermodynamic effects on the protein are complicated by additional factors. In particular, the effects of cysteine substitutions on the native state and the stabilization of compact nonnative states by the disulfide can override any stabilizing effect of the cross-link. PMID- 10794422 TI - Structure of an analog of fusion peptide from hemagglutinin. AB - A 20-residue peptide E5 containing five glutamates, an analog of the fusion peptide of influenza virus hemagglutinin (HA) exhibiting fusion activity at acidic pH lower than 6.0-6.5 was studied by circular dichroism (CD), Fourier transform infrared, and 1H-NMR spectroscopy in water, water/trifluoroethanol (TFE) mixtures, dodecylphosphocholine (DPC) micelles, and phospholipid vesicles. E5 became structurally ordered at pH < or = 6 and the helical content in the peptide increased in the row: water < water/TFE < DPC approximately = phospholipid vesicle while the amount of beta-structure was approximately reverse. 1H-NMR data and line-broadening effect of 5-, 16-doxylstearates on proton resonances of DPC bound peptide showed E5 forms amphiphilic alpha-helix in residues 2-18, which is flexible in 11-18 part. The analysis of the proton chemical shifts of DPC bound and CD intensity at 220 nm of phospholipid bound E5 showed that the pH dependence of helical content is characterized by the same pKa approximately 5.6. Only Glu11 and Glu15 in DPC bound peptide showed such elevated pKas, presumably due to transient hydrogen bond(s) Glu11 (Glu15) deltaCOO- (H+)...HN Glu15 that dispose(s) the side chain of Glu11 (Glu15) residue(s) close to the micelle/water interface. These glutamates are present in the HA-fusion peptide and the experimental half-maximal pH of fusion for HA and E5 peptides is approximately 5.6. Therefore, a specific anchorage of these peptides onto membrane necessary for fusion is likely driven by the protonation of the carboxylate group of Glu11 (Glu15) residue(s) participating in transient hydrogen bond(s). PMID- 10794423 TI - Folding of a dimeric beta-barrel: residual structure in the urea denatured state of the human papillomavirus E2 DNA binding domain. AB - The dimeric beta-barrel is a characteristic topology initially found in the transcriptional regulatory domain of the E2 DNA binding domain from papillomaviruses. We have previously described the kinetic folding mechanism of the human HPV-16 domain, and, as part of these studies, we present a structural characterization of the urea-denatured state of the protein. We have obtained a set of chemical shift assignments for the C-terminal domain in urea using heteronuclear NMR methods and found regions with persistent residual structure. Based on chemical shift deviations from random coil values, 3'J(NHN alpha) coupling constants, heteronuclear single quantum coherence peak intensities, and nuclear Overhauser effect data, we have determined clusters of residual structure in regions corresponding to the DNA binding helix and the second beta-strand in the folded conformation. Most of the structures found are of nonnative nature, including turn-like conformations. Urea denaturation at equilibrium displayed a loss in protein concentration dependence, in absolute parallel to a similar deviation observed in the folding rate constant from kinetic experiments. These results strongly suggest an alternative folding pathway in which a dimeric intermediate is formed and the rate-limiting step becomes first order at high protein concentrations. The structural elements found in the denatured state would collide to yield productive interactions, establishing an intermolecular folding nucleus at high protein concentrations. We discuss our results in terms of the folding mechanism of this particular topology in an attempt to contribute to a better understanding of the folding of dimers in general and intertwined dimeric proteins such as transcription factors in particular. PMID- 10794424 TI - Scoring functions in protein folding and design. AB - We present an analysis of the assumptions behind some of the most commonly used methods for evaluating the goodness of the fit between a sequence and a structure. Our studies on a lattice model show that methods based on statistical considerations are easy to use and can capture some of the features of protein like sequences and their corresponding native states, but unfortunately are incapable of recognizing, with certainty, the native-like conformation of a sequence among a set of decoys. Meanwhile, an optimization method, entailing the determination of the parameters of an effective free energy of interaction, is much more reliable in recognizing the native state of a sequence. However, the statistical method is shown to perform quite well in tests of protein design. PMID- 10794425 TI - The sarcosine effect on protein stability: a case of nonadditivity? AB - We have used differential scanning calorimetry to determine the effect of low concentrations (C = 0-2 M) of the osmolyte sarcosine on the Gibbs energy changes (deltaG) for the unfolding of hen-egg-white lysozyme, ribonuclease A, and ubiquitin, under the same buffer and pH conditions. We have also computed this effect on the basis of the additivity assumption and using published values of the transfer Gibbs energies for the amino acid side chains and the peptide backbone unit. The values thus predicted for the slope delta deltaG/deltaC agree with the experimental ones, but only if the unfolded state is assumed to be compact (that is, if the accessibility to solvent of the unfolded state is modeled using segments excised from native structures). The additivity-based calculations predict similar delta deltaG/deltaC values for the three proteins studied. We point out that, to the extent that this approximate constancy of delta deltaG/deltaC holds, osmolyte-induced increases in denaturation temperature will be larger for proteins with low unfolding enthalpy (small proteins that bury a large proportion of apolar surface). The experimental results reported here are consistent with this hypothesis. PMID- 10794426 TI - Immunoglobulin class switch recombination machinery: progress and challenges. PMID- 10794427 TI - A locus closely linked to Mtv7 on mouse chromosome 1 influences development of acute versus chronic graft-versus-host disease in a murine model. AB - The relationship between acute and chronic graft-versus-host disease (GVHD) is not well understood. A murine model of acute and chronic GVHD is the B6D2F1 parent-->F1 model in which transfer of C57BL/6 parental strain lymphoid cells to B6D2F1 recipients results in development of Th1-mediated acute GVHD, whereas transfer of DBA/2 parental strain lymphoid cells to B6D2F1 recipients results in development of Th2-mediated chronic GVHD. Numerous studies have investigated the reason for the differential development of acute versus chronic GVHD in this model but have as yet failed to identify the factor that determines which type of T helper cell will predominate and thereby which type of GVHD will develop. In this report, we demonstrate, using congenic strains of mice, that a locus in the vicinity of the Mtv7 locus on Chromosome 1 of the mouse significantly influences development of acute versus chronic GVHD in the B6D2F1 model. PMID- 10794428 TI - Correlation of loss of CD4 T cells with plasma levels of both soluble form Fas (CD95) Fas ligand (FasL) in HIV-infected infants. AB - Fas and Fas ligand (FasL), members of the TNFR and TNF families of molecules involved in apoptosis, respectively, are expressed in membrane-associated as well as soluble forms. Soluble Fas (sFas) and sFasL were evaluated in sequential samples from 16 HIV-infected and 11 HIV-exposed uninfected infants at ages 0-13 months. Regardless of the state of infection, age-dependent decreases in peripheral CD4 T cell counts and increases in sFas and sFasL were noted. However, decreases of the percentage CD4 T cells were more prominent in HIV-infected infants, and this was correlated significantly with increased plasma levels of sFas and sFasL (P = 0.002 and 0.004, respectively). Moreover, the levels of sFas in HIV-infected infants were found to be directly correlated with plasma HIV RNA (P = 0.03) and were significantly increased as early as age <1 month and prior to the onset of CD4 T cell decline. In uninfected infants, there was no such correlation between CD4 counts and the levels of sFas/sFasL. Plasma levels of sFas and sFasL may thus be important indicators of disease progression in perinatal HIV infection. PMID- 10794429 TI - Fab fragments directed against laminin 5 induce subepidermal blisters in neonatal mice. AB - Patients with one form of cicatricial pemphigoid have IgG autoantibodies directed against laminin 5 (alpha3beta3gamma2), an adhesion protein in epidermal basement membrane. Anti-laminin 5 autoantibodies are not found in patients with other skin or mucosal diseases and hence serve as a specific marker for this autoimmune blistering disorder. The demonstration that experimental and patient anti-laminin 5 IgG are pathogenic in animal models indicated that such autoantibodies are central to disease pathophysiology. To investigate further the role of antibody valence and complement in triggering lesion formation in vivo, rabbit anti laminin 5 (or normal, control) Fab fragments were passively transferred to neonatal BALB/c mice. Mice receiving anti-laminin 5 Fab fragments developed, in a dose-related fashion, circulating anti-basement membrane antibodies, deposits of immunoreactive rabbit IgG (but not murine C3) in epidermal basement membranes, and subepithelial blisters of skin and mucous membranes. Such alterations were not observed in mice treated with equivalent concentrations of normal rabbit Fab fragments. These studies demonstrated that neither complement activation nor cross-linking of laminin 5 in epidermal basement membranes was required for induction of subepidermal blister formation in this animal model of a human autoimmune bullous disease. PMID- 10794430 TI - Efficient detection of thirty-seven new IL2RG mutations in human X-linked severe combined immunodeficiency. AB - X-linked severe combined immunodeficiency (XSCID) is a rare and potentially fatal disease caused by mutations of IL2RG, the gene encoding the interleukin-2 receptor gamma chain, a component of multiple cytokine receptors that are essential for lymphocyte development and function. To date, over 100 different mutations of IL2RG resulting in XSCID have been published. Using nonradioactive, direct DNA sequencing of a single PCR amplicon containing the whole IL2RG gene, we found IL2RG mutations in 78 previously unpublished unrelated cases of XSCID. We report 37 newly identified mutations of IL2RG, including 23 point mutations, 10 small deletions, 3 instances of the same single nucleotide insertion, 1 large deletion, and 2 complex mutations. More than half of the mutations (22 of 37) were predicted to result in unstable IL2RG mRNA. The remaining 14 mutations disrupted conserved functional motifs common to all cytokine receptor family members; changed protein conformation, charge, or hydrophobicity; or altered the intracellular portion of the protein, which is critical for proper interaction with signal-transducing molecules including Janus family tyrosine kinase 3. PMID- 10794431 TI - Development of autologous T lymphocytes in two males with X-linked severe combined immune deficiency: molecular and cellular characterization. AB - We report on two patients affected with severe combined immune deficiency (SCID) with an unusual immunological phenotype and a substantial number of autologous, poorly functioning T cells. Distinct mutations identified at the IL2RG locus in the two patients impaired IL-2-mediated signaling but affected T-cell lymphopoiesis differently, resulting in generation of a polyclonal or oligoclonal T-cell repertoire. These observations add to the growing complexity of the immunological spectrum of SCID in humans and indicate the need for detailed immunological and molecular investigations in atypical cases. PMID- 10794432 TI - Eotaxin and RANTES expression by the dermal endothelium is associated with eosinophil infiltration after ivermectin treatment of onchocerciasis. AB - The roles of eotaxin, RANTES, and MCP-3 expression in eosinophil recruitment to the site of parasite killing that occurs following ivermectin treatment of onchocerciasis were assessed in the skin of 13 Onchocerca volvulus-infected subjects and two noninfected controls before and after ivermectin treatment. Adverse reactions in infected subjects were associated with the appearance of eosinophils in the dermis as part of a perivascular inflammatory infiltrate. Although no expression of RANTES and eotaxin was seen in dermal vascular endothelial cells in biopsies taken before treatment (nor at any time in the skin of uninfected controls), endothelial expression of both eotaxin and RANTES was noted by 24 h following treatment. While RANTES expression was transient, eotaxin expression increased in parallel with increasing eosinophil recruitment up to 60 h posttreatment. These observations indicate that endothelial expression of eotaxin and RANTES may have an important role in eosinophil recruitment into the skin during helminth-killing reactions. PMID- 10794433 TI - Partial impairment of immune functions in peripheral blood leukocytes from aged men with Down's syndrome. AB - Down's syndrome (DS) has been considered a model of accelerated aging and of Alzheimer's disease. We investigated immunologic functions using peripheral blood leukocytes in order to correlate the production of cytokines and development of neuropathological changes of Alzheimer type in aged persons with DS. Cytokine production (IL-1beta, IL-2, IL-6, IL-8, and TNF-alpha), phytohemagglutinin (PHA) stimulated proliferation of nonadherent monocytes, and superoxide anion production from polymorphonuclear leukocytes were measured. PHA-stimulated proliferation in aged individuals (>30 years old) with DS was significantly lower than that of age- and sex-matched controls (DS vs control, 55,707+/-5810 vs 88,310+/-6994 cpm, P < 0.001). PHA-stimulated IL-2 production was also significantly decreased in aged individuals with DS (DS vs control, 7.1+/-2.1 vs 10.7+/-1.3 ng/ml). Interestingly, the decrease of proliferation and IL-2 production in aged males with DS is significantly greater than in aged women with DS. PHA-stimulated proliferation and IL-2 production of nonadherent monocytes in females was not significantly reduced. IL-1beta production by LPS-activated adherent monocytes was significantly decreased in older adults with DS compared with non-DS controls. Other immune parameters measured in DS were not significantly different from that of age-matched controls. We conclude that there is partial impairment of T lymphocytes in aged persons with DS that is significantly greater in males than in females. PMID- 10794434 TI - The complexicity of cytokine treatment in ongoing EAE induced with MBP peptide 68 86 in Lewis rats. AB - IL-10 and TGF-beta1 are important immunoregulatory cytokines associated with clinical remissions in multiple sclerosis and amelioration of experimental allergic encephalomyelitis (EAE). IL-10 and TGF-beta1 have previously been shown to prevent the development of EAE. Here, we study effects of IL-10 and TGF-beta1 in ongoing EAE. When IL-10 or TGF-beta1 was administered by the nasal route from day 0 to day 7 postimmunization (pi), both IL-10 and TGF-beta1 prevented the development of acute EAE in Lewis rats. When IL-10 or TGF-beta1 was administered by the nasal route from day 5 to day 12 pi, both IL-10 and TGF-beta1 failed to influence clinical EAE. The inhibition of clinical EAE severity in IL-10 prevented rats was associated with reduced proliferation, IFN-gamma mRNA expression, and IFN-gamma secretion, while proliferation as well as IFN-gamma mRNA expression and secretion were augmented in TGF-beta1-prevented rats. TGF beta1-prevented rats exhibited high levels of NO production by DC, which may mediate apoptosis of CD4+ T cells and of the DC themselves. For prevention, both IL-10 and TGF-beta1 inhibited infiltration of CD4+ T cells within the CNS, but neither IL-10 nor TGF-beta1 induced immune deviation from Th1 to Th2. Expression of IL-4 mRNA was not altered in IL-10- and TGF-beta1-prevented rats. These results demonstrate that IL-10 and TGF-beta administration by the nasal route can prevent the development of acute EAE, but by different mechanisms. The findings in rats with ongoing EAE have implications for the clinical application of cytokine treatment in autoimmune diseases. PMID- 10794435 TI - On the uniqueness of the positive solution of an integral equation which appears in epidemiological models. AB - In this paper, we show that the positive solution of a nonlinear integral equation which appears in classical SIR epidemiological models is unique. The demonstration of this fact is necessary to justify the correctness of any approximate or numerical solution. The SIR epidemiological model is used only for simplicity. In fact, the methods used can be easily extended to prove the existence and uniqueness of the more involved integral equations that appear when more biological realities are considered. Thus the inclusion of a latent class (SLIR models) and models incorporating variability in the infectiousness with duration of the infection and spatial distribution lead to integral equations to which the results derived in this paper apply immediately. PMID- 10794436 TI - Dynamics of variable-yield nutrient-phytoplankton-zooplankton models with nutrient recycling and self-shading. AB - Several nutrient-phytoplankton-zooplankton models with internal nutrient storage by phytoplankton are derived and analyzed. It is shown that there are thresholds beyond which the system is uniformly persistent. Variable-yield models with self shading of phytoplankton are also considered. With respect to uniform persistence, our result demonstrates that the global dynamics of the system with shading are the same as those for which the self-shading mechanism is ignored. PMID- 10794437 TI - Protected polymorphism in the two-locus haploid model with unpredictable fitnesses. AB - In an unpredictable environment, the distributions of alleles from which polymorphism can be maintained forever belong to a certain set, the C-viability kernel. Such a set is calculated in the two-locus haploid model, as well as the corresponding fitnesses at any time which make this maintenance possible. The dependence of the C-viability kernel on the set U of admissible fitnesses and on the recombination rate r is studied. Notably, the C-viability kernel varies rapidly in the neighborhood of equal fitness of AB and ab; it becomes empty when ab has a fitness below a certain function, which is delineated, of the recombination rate. The properties of the two-locus model under constraints, out of equilibrium and with unpredictable selection are thus presented. PMID- 10794438 TI - Random binding of dimers to chains. AB - We develop a probabilistic model for the binding of a small linear polymer to a larger chain. We assume that we can approximate the energy of interaction of the two chains by summing the pairwise interactions between subunits. Because the energy of interaction between a pair of subunits can depend on neighboring subunits, which we assume vary along the chain, we assign the pairwise energies of interactions according to a specified probability distribution. Thus we develop a statistical model for the binding of two molecules. While such models may not be appropriate for studying the interaction of a particular pair of molecules, they can provide insight into questions that deal with populations of molecules, such as why do MHC molecules bind peptides of a certain size? Here we analyze in detail the special case of a heterodimer binding to a polymer. PMID- 10794439 TI - Intramolecular charge transfer at reverse micelle-water pool interface: p-N,N dimethylaminobenzoic acid in AOT/cyclohexane/water reverse micelle. AB - Photoinduced intramolecular charge transfer (ICT) of p-N,N-dimethylaminobenzoic acid (DMABOA) in AOT/cyclohexane/H2O reverse micelle was investigated and compared with that in CTAB/1-heptanol/H2O reverse micelle. It is proposed that the DMABOA molecule exists at the AOT reverse micelle water pool interface with its carboxylic group heading toward the water pool while the dimethylaminophenyl moiety buried in the micellar phase. Dual fluorescence of DMABOA that is indicative of the ICT reaction in the excited state was observed over the investigated water pool size, W of 3-17, in the AOT reverse micelle. The ICT emission of DMABOA in the AOT reverse micelle-water pool interface was found to be much weaker than that in the CTAB reverse micelle-water pool interface, and was attributed to the parallel direction of the electric field at the AOT reverse micelle-water pool interface to the charge transfer. PMID- 10794440 TI - Heteronuclear diatomic force constants clarified through perturbation theory II. AB - A simple relationship between the heteronuclear diatomic force constant (K(AB)) and the homonuclear diatomic force constants (K(AA), K(BB)), which was proposed in a previous report, has been improved through the second-order perturbation theory as K(AB) = zeta3(K(AA) x K(BB))(1/2); zeta = (R(AA) x R(BB))(1/2)/R(AB) where zeta denotes the correction factor in which R(AB), R(AA), and R(BB) are the equilibrium internuclear distances of diatomic molecules AB, AA, and BB, respectively. To test the above expression, a large number of heteronuclear diatomic force constants have been calculated and compared with those obtained from normal coordinate analyses as well as ab initio quantum mechanical methods (Gaussian 98W). We have found that the above modified expression better reproduces the force constants of most heteronuclear diatomic molecules than the previous expression. It is therefore expected that the expression may also be applied to the prediction of stretching force constants between heteronuclear diatomics in various polyatomic molecules. PMID- 10794441 TI - Principal component analysis of the absorption and resonance Raman spectra of the metallochromic indicator antipyrylazo III. AB - Metallochromic indicators, whose spectral properties are changed in the presence of metal cations, are used mainly in biological studies to monitor Ca2+ and Mg2+ ions. Antipyrylazo III is such indicator, employed for mid-range Ca2+ concentrations (10-1000 microM). The stoichiometry of the interactions of antipyrylazo III with Ca2+, Mg2+, Ba2+, Sr2+ and Zn2+ ions and the relevant binding constants were studied by principal component analysis (PCA) of the absorption spectral changes. The resonance Raman spectra of the above systems were measured as well, and the resolved Raman spectra of the various species were calculated and assigned. The vibrational spectra are more featured, more characteristic of the binding ions and exhibit stronger relative spectral changes upon binding the cations. The basis sets of Raman spectra could thus be used as an analytical tool for these divalent metallic cations. PMID- 10794442 TI - A vibrational study of di-i-propoxyphosphoryl benzylisothiourea. AB - The Fourier transform infrared and Raman spectra of di-i-propoxyphosphoryl benzylisothiourea (DPB) (1) in the solid state and in solutions of CCl4, CHCl3, CHBr3, CH2Cl2, C2H4Cl2, C2H4Br2 and THF were studied. In the IR spectra, the effects of different concentrations were also investigated. The behavior of the nu(NH), delta(NH), delta(HNH), nu(C=N) and nu(P=O) normal modes suggests the existence of a tautomerism between the phosphorylamine (I) and N-phosphorylimine (II) structures: [structures: see text] The data show the presence of different delta(NH) and delta(HNH) bendings and nu(C=N) normal modes in the solid state as a result of inter and intramolecular hydrogen bonding. The experimental approximate frequencies assignments were done for this compound, and were confirmed by a normal coordinate analysis carried out for several fragments of phosphorylamine and N-phosphorylimine structures. PMID- 10794444 TI - Emission spectra of HCN/HNC in the 2-5 microm range of astrophysical interest. AB - Emission from H12C14N observed with a Fourier transform spectrometer from a radio frequency excited plasma is reported in the 2400-3400 cm(-1) spectral range of astrophysical interest. The molecular constants, for 21 vibration-rotation bands are given, as well as estimates of the first-order Herman-Wallis coefficients for 11 bands. These constants are derived from about 900 observed transitions in HCN, and are used to generate a sequential linelist of about 1400 calculated line positions, within a standard deviation equal to 3 x 10(-4) cm(-1). The relative intensities of the observed lines are also reported, as well as those for the nu1 band of H14N12C, at 3650 cm(-1), simultaneously observed from the same plasma. PMID- 10794443 TI - Ab initio studies on the vibrational spectra and conformational isomerism of 2,4 dimethyl-1,3-pentadiene. AB - Ab initio was used to study the structure of various conformational isomers and their vibrational spectra of 2,4-dimethyl-1,3-pentadiene (2,4-PD) in detail. Two stable conformations, s-trans and s-cis, were found in which s-trans is more stable. The geometry of stable conformations and charge distributions were studied, and the effect of different basis sets on geometry optimization is discussed. The results of complete optimization indicate that molecular skeleton is nearly in a plane, its largest deviation is only 0.3 degrees. Therefore, it is reasonable and available to hypothesis that the molecule has Cs symmetry. The thermal dynamics conformations were calculated and compared with experimental values. DeltaH(o) between two conformations of 2,4-PD measured from experiment is 4.36 kJ/mol, deltaS(o) is 2.56 J/mol K., calculated results are slightly different from experimental ones. Vibrational frequencies of 2,4-PD conformers have been studied by ab initio molecular orbital calculations using different basis set. The calculated vibrational frequencies are analyzed and compared with the experimental spectra. PMID- 10794445 TI - Investigation of chelate formation, intramolecular energy transfer and luminescence efficiency and lifetimes in the Eu-thenoyltrifluoroacetone trioctylphosphine oxide-Triton X-100 system using absorbance, fluorescence and photothermal measurements. AB - The formation and the photophysical properties of the europium thenoyltrifluoroacetone (TTA) trioctylphosphine oxide (TOPO)-Triton X-100 chelate were investigated. When the medium is buffered with acetate, there is a strong competition between acetate and TTA for coordination with europium ions. When TOPO is added into the solution, the Eu-TTA-TOPO ternary chelate forms more easily, probably because the coligand acts as a synergic agent and would favour the formation of the enol form of TTA. Although the stoichiometric composition of the chelate is expected to be Eu(TTA)3(TOPO)2, the Eu-TTA and the Eu-TOPO mole ratios may be within 2-3 and 1-2, respectively, depending on the composition of the solution. However, the fluorescent properties of the chelate seem to be mainly dominated by its actual concentration into the solution rather than by its composition. Time resolution of europium emission spectra in the microsecond range has shown that energy transfer occurs from the TTA ligand to the 5D1 level of europium. Then, the emitting 5D0 level is populated through non-radiative deactivation of 5D1. The observed lifetimes of the 5D1 and 5D0 states are 1.25 and 860 micros, respectively. The overall fluorescence quantum yield of the chelate, measured by the photothermal method, is found to be 0.22. On the basis of the time-resolved photothermal experiments, the fluorescence quantum yield of the 5D0 state is expected to be > 0.8 and the energy transfer efficiency < 0.28. PMID- 10794446 TI - UV-vis and Raman spectroelectrochemical investigation of the redox behavior of poly(5-cyanoindole) in acidic aqueous solutions. AB - Spectroelectrochemical properties of conducting poly(5-cyanoindole) films deposited on indium tin oxide (ITO) and platinum electrodes are investigated using UV-vis and resonant Raman spectroscopies. The transitions from undoped to semi-conducting state of P5CN require the partial oxidation of the polymer to create radical-cations by insertion of charge-neutralizing anions into the polymer. In order to obtain detailed structural information from the vibrational spectra, it is necessary to know the vibrational modes of oxidation-sensitive bands. Vibrational assignments were made on the basis of the results obtained on polyindole and P5CN in acetonitrile solution. The drastic changes in optical absorption and Raman spectra observed at various stage of oxidation were explained by the conversions between at least three different structures. On the basis of the Raman spectra, we have identified the vibrational modes associated with neutral and polaronic segments. The perturbation associated with the coexistence of these polaronic segments has been described as a quinoid structure growing on the expense of the benzoid one. The results obtained indicate that the molecular properties of the conducting polymers at various stages of an oxidation are better revealed by in-situ Raman spectra than by ex-situ studies. PMID- 10794448 TI - Internal axis molecular parameters, torsional energies and matrix elements of methyl mercaptan-D2. AB - In this paper an internal axis method Hamiltonian model has been applied to evaluate the torsional rotational molecular parameters of asymmetrically substituted methyl mercaptan (CHD2SH) using previously observed microwave transitions. The torsional potential barrier function V2 has been obtained. The pure torsional energies and matrix elements between various torsional sub-levels up to the fourth excited torsional state in the ground vibrational state have been determined. The matrix elements and the torsional energies will be of great value to researchers seeking the spectrum of this molecule. PMID- 10794447 TI - Multivariate analysis and quantitation of (17)O-nuclear magnetic resonance in primary alcohol mixtures. AB - Multivariate techniques were used to address the quantification of (17)O-nuclear magnetic resonance (NMR) spectra for a series of primary alcohol mixtures. Due to highly overlapping resonances, quantitative spectral evaluation using standard integration and deconvolution techniques proved difficult. Multivariate evaluation of the (17)O-NMR spectral data obtained for 26 mixtures of five primary alcohols demonstrated that obtaining information about spectral overlap and interferences allowed the development of more accurate models. Initial partial least squares (PLS) models developed for the (17)O-NMR data collected from the primary alcohol mixtures resulted in very poor precision, with signal overlap between the different chemical species suspected of being the primary contributor to the error. To directly evaluate the question of spectral overlap in these alcohol mixtures, net analyte signal (NAS) analyses were performed. The NAS results indicate that alcohols with similar chain lengths produced severely overlapping (17)O-NMR resonances. Grouping the alcohols based on chain length allowed more accurate and robust calibration models to be developed. PMID- 10794449 TI - A study by ultraviolet spectroscopy on self-association of purine, 6 methylpurine, benzimidazole, and imidazo [1,2-a]pyridine in aqueous solution. AB - The self-association of four heteroaromatic compounds with fused five- and six membered rings: purine, 6-methylpurine, benzimidazole, and imidazo[1,2 a]pyridine, was studied in aqueous solution at different pH values, by ultraviolet spectroscopy. The variation in molar absorptivity with the concentration of these compounds was measured for the band systems in the mid ultraviolet region. Hypochromic deviations from the Beer-Lambert law with increasing concentration were found for all compounds. These results were interpreted in terms of self-association processes involving the formation of dimers and, in certain cases, also the formation of polymers. From the fitting of the experimental curves of hypochromic effects, self-association constants for dimerization and polymerization were calculated. The results obtained are discussed in relation to the self-association properties of the building blocks of each compound, studied by us in previous works. The relevance of the position of the nitrogen atoms at the five- and six-membered rings was analyzed to elucidate the nature of the self-association processes, in particular the tendency to polymerization. The values of the calculated self-association constants are discussed in relation to the intermolecular interaction energies of these compounds in aqueous solution. PMID- 10794451 TI - Transition probabilities and average cross sections for the Na2 B1pi(u) --> X1sigma(g)+ system using as excitation the 4880 A Ar+ laser line. AB - The fluorescence spectrum of Na2 induced by the 4879.86 A line of an Argon ion laser has been analyzed with special emphasis on determination of accurate relative intensities. We have observed nineteen fluorescence series for the B1pi(u) --> X1sigma(g)+ band system. Some series are reported for the first time. The radiative transition probabilities for the observed fluorescence series were calculated using hybrid potential energy curves for the B1pi(u) and X1sigma(g)+ states constructed up to dissociation and a B-X transition dipole moment function. Radiative lifetimes for the rovibrational levels of the upper states pumped by the laser line have also been calculated. The transition probabilities and lifetimes agree with the corresponding observed measurements usually within the experimental uncertainty. From the rotational satellite structure with deltaJ' = +/- 1, +/- 2...+/- 20, for some nu''-bands of the most intense fluorescence series induced by emission from the vibrational-rotational levels: nu' = 6, J' = 43 and v' = 9, J' = 56, collision-induced transition rates and average cross sections have been obtained. PMID- 10794450 TI - Detection of CN radicals in DC nitrogen plasma used for deposition of CNx layers. AB - Emission spectra from a DC plasma discharge of nitrogen with a graphite cathode used for deposition of CNx layers were investigated in the visible range 350-900 nm. The spectra recorded at low and high resolution from both the negative glow and the positive column of the discharge were studied separately. All spectra are dominated by neutral and ionised N2 emission. In the positive column the violet band of the cyanogen (CN) radical was identified and analysed for vibrational structure. From a computer simulation of the rotationally resolved violet band, vibrational temperatures were derived and found to be in the intensity distribution for the nu = 0, 1 and 2 levels from thermal equilibrium. In the negative glow the strong N2+ features completely mask the spectral region of the violet band of CN. Conclusions were drawn concerning the CN formation by chemical sputtering, and the role of CN radicals in the formation of polymeric CNx layers of 1:1 = C:N stoichiometry. PMID- 10794452 TI - Systematic investigation of absorption, fluorescence and laser properties of some p- and m-oligophenylenes. AB - Absorption, fluorescence and laser properties of ten selected aromatic compounds from the oligophenylene family are studied experimentally at room temperature (293 K). The first eight compounds are arranged in such way that odd numbered compounds reveal 1Lb --> 1A fluorescence, while even numbered compounds show 1La -> 1A fluorescence. All compounds are family related in pi-structure and are of the same degree of planarity and rigidity. The quantum yield of fluorescence, gamma, and the decay times, tau(f), of non-deaerated and deaerated cyclohexane solutions are measured. The oscillator strength, f(e), the fluorescence rate constant, Kf, natural lifetimes, tau(o)T, and intersystem crossing rate constant, K(ST), are calculated. The lowest 1Lb and 1La singlet and 3La, triplet (77 K) levels are determined. Investigations showed that transition from a polyphenyl molecule which shows 1La --> 1A fluorescence to a family related in the pi structure molecule which reveals the 1Lb --> 1A fluorescence is accompanied by certain changes in all the fluorescence parameters. This indicates that gamma decreases, tau(f) increases, Kf and the FWRE of the fluorescence spectrum decrease. Moreover, K(ST) also decreases, sometimes very significantly. The decrease in the K(ST) value is explained by the fact that matrix elements of the spin-orbit coupling of the S alpha and Ti states are much lower in value than analogous elements of the spin-orbit coupling of Sp and Ti states. It is shown that all p-polyphenyles exhibit excellent laser action, while m-polyphenyles do not produce laser oscillation under any conditions. The values of K(ST) and other fluorescence parameters measured can be used for various practical purposes and theoretical considerations. PMID- 10794453 TI - Energy transfer in triton-X 100 micelles: a fluorescence study. AB - The study of fluorescence energy transfer from the phenyl groups of the micellar triton X-100 (TX-100) to solubilised 1-pyrene butyric acid (PBA) has been carried out. Through the analysis of the donor fluorescence quenching energy transfer efficiency has been determined. The observed donor-acceptor separation suggests that pyrene molecules are distributed uniformly in the micellar core. PMID- 10794454 TI - Interaction of cyanine dyes with nucleic acids. XVII. Towards an aggregation of cyanine dyes in solutions as a factor facilitating nucleic acid detection. AB - Spectral properties of newly synthesized cyanine dyes, namely 1-[6-(4-[6-[2,6 dimethyl-4-(3-methyl-2,3-dihydro-1,3-benzothiazol- 2-ylidenmethyl)-1 pyridiniumyl]hexanoyl]piperazino)-6- oxohexyl]-2,6-dimethyl-4-(3-ethyl-2,3 dihydro-1,3-benzothiazol+ ++-2-ylidenmethyl)pyridinium (K-6) (bichromophoric dye) and 1-[5-di(3-[5-[2,6-dimethyl-4-(3-methyl-2,3-dihydro-1,3-benzothiazol++ +-2 ylidenmethyl)-1-pyridiniumyl]pentylcarboxamido]pro pyl) carbamoylpentyl]-2,6 dimethyl-4-(3-methyl-2,3-dihydro-1,3-benzo thiazol-2-ylidenmethyl) pyridinium (K T) (trichromophoric dye) in solutions in the presence of and without deoxyribonucleic acid (DNA) were studied within a wide concentration range. It has been established that absorption, as well as fluorescence of investigated dye solutions, without DNA are mainly determined by H-aggregates of dye molecules. On the contrary, the fluorescence of dye solutions in the presence of DNA gives an intrinsic dye molecular fluorescence. H-aggregates are broken because of binding dye molecules with DNA. It has been suggested that both K-T and K-6 molecules bind mainly with DNA via the interaction of two chromophores. As the ratio of the number of dye molecules to that of DNA base pairs increases with an increase in dye concentration, a formation of dye molecule H-aggregates on DNA molecules are observed. Such aggregates have a different structure than those formed in the solutions without DNA. On the grounds of the data obtained, it is concluded that it is possible to use a dye aggregation capable of obtaining higher values for fluorescence enhancement of the DNA stains. PMID- 10794455 TI - Speciation and structure of copper(II) complexes with histidine-containing peptides in aqueous medium: a combined potentiometric and spectroscopic study. AB - The complex formation between copper(II) and some histidine-containing peptides has been investigated by means of potentiometric and spectroscopic measurements. Due to the interesting co-ordination mode towards copper(II), peptides with histidyl residue located in second position from the N-terminal amino group have been chosen. The stability constants evaluation has been performed by both pH metric and ESR (room temperature) measurements; in this context a suitable computer program for the calculation of both stability constant values and the ESR spectrum for each complex has been written. Visible spectrophotometric and circular dichroism spectra, together with the isotropic ESR parameters, were used in order to propose a structure for each complex having a significant percentage of formation in solution. PMID- 10794456 TI - Conformational and spectroscopic investigation of 3-hydroxyflavone-aluminium chelates. AB - 3-Hydroxyflavone (3HF), which is the simplest molecule of the flavonol class, possesses chelating properties towards Al(III). Spectrophotometric methods have shown that the 3HF molecule forms an Al(3HF)2 complex in pure methanol. The structure of this complex, obtained by quantum semi-empirical AM1 method, indicated that complexed 3HF adopts a pyronium form. Structural and electronic modifications induced by chelation are illustrated by the important frequency shifts observed between free and complexed 3HF FT-Raman spectra and by the chemical shifts variations in the 13C NMR spectra of the two species. Complexes with the same stoichiometry were formed when AcO- or MeO- are present in the medium. However, in acidic medium the chelate composition is Al2(3HF). PMID- 10794457 TI - Structural characterization of glassy phases in the system Na2O-Ga2O3-P2O5 by MAS NMR, EXAFS and vibrational spectroscopy. I. Cations coordination. AB - The cationic coordinations of phosphate based gallium sodium glasses in the system Na2O-Ga2O3-P2O5 have been studied by several techniques (71Ga and 23Na MAS NMR, EXAFS and vibrational spectroscopies) in order to study the relationship between the structure and the chemical composition. We found that three different environments are available for the gallium ions while it is very difficult to get accurate information on the sodium coordinations. Our data show that in orthophosphate glasses, gallium is mainly tetrahedral but when the mean phosphate chain length increases, its coordination becomes more and more octahedral. In these glassy structure, it becomes then possible to dissolve large amounts of typically octahedral cations like Fe3+ or Cr3+. PMID- 10794458 TI - Structural characterization of glassy phases in the system Na2O-Ga2O3-P2O5 by MAS and solution NMR and vibrational spectroscopy: II. Structure of the phosphate network. AB - The structure of the phosphate network of glasses in the system Na2O-Ga2O3-P2O5 has been investigated as a function of the Na/Ga molar ratio and the phosphate composition corresponding to a mono, di, tri, tetra and meta phosphate stoichiometry. The glass is made of phosphate molecular groups of different lengths linked by the cations with rather ionic (Na) or more covalent (Ga) bonds except in the case of the orthophosphate composition for which we only found isolated PO4(3-) ions. The vibrational spectra are sensitive to composition variations but the band width and the couplings between the different groups prevent any quantitative determination. 31P MAS-NMR gives an in-situ information on the P environment in the glass but the signals are often large and ill defined so that the assignment is not at all straightforward. On the other hand, 31P solution NMR gives sharper signals, allowing more quantitative determinations but the dissolution process always introduces some indeterminacy on the real glass structure which can be minimized by a careful preparation of the solution. PMID- 10794459 TI - H-bonding effects on the IR and NMR spectra of N-tosyl-amino acid 2,6 bishydroxylmethyl pyridine monoesters. AB - H-bonding in the title monoesters affects the most utilized types of spectrometries-infrared (IR) and nuclear magnetic resonance (NMR). The shifts of the three kinds of A-H stretching bands to lower wavenumbers in the vibrational spectra, and the 1H chemical shifts, which are to the low field, are observed and reasonably explained in this paper. PMID- 10794460 TI - Pressure-tuning infrared and Raman spectroscopy of group 14 tetraphenyl compounds. AB - The effect of high external pressures on the vibrational spectra of the tetraphenyl Group 14 compounds, Ph4M (M = Si, Ge, Sn, Pb), were examined between ambient pressure and 40 kbar with the aid of a diamond-anvil cell. The four compounds displayed similar behaviour as the pressure was increased and a structural transition at approximately 15 kbar, most probably associated with a phenyl ring rotation, was identified in each case. The pressure dependencies of selected vibrational modes were obtained. PMID- 10794461 TI - A modified diffuse reflectance infrared Fourier transform spectroscopy cell for depth profiling of ceramic fibers. AB - A modified diffused reflectance infrared Fourier transformed spectroscopy (DRIFTS) accessory was used to analyze the surface properties of alumino-silicate fibers. The modifications are simple and involve a different way of performing depth-profiling from traditional DRIFTS by removing approximately 2 mm of salt from a full cup prior to placing the sample in for depth profiling. This method proved successful in elucidating the effects of quenching alumino-silicate fibers in mineral oil versus quenching in an air stream. PMID- 10794462 TI - Spectroscopic studies of the Eu3+ and Er3+ ions in the fluorozirconate LaZr2F11 matrix. AB - An investigation by optical spectroscopy of the Eu3+ and Er3+ active ions in the crystallized fluorozirconate matrix LaZr2F11 is presented. The 5D1-->7F0-5 emission lines of Eu3+ are used to extract the 7F0-5 energy scheme and the observed extinctions permit the deduction of irreducible representations (IRREPS) associated with corresponding sub-levels in the D2 symmetry. The crystal field analysis was carried out on a 387 x 387 basis set, comprising the 7F, 5D(1,2,3) 5F(1,2), 5G(1,2,3) and 3P(1,2,3,4,5,6) terms of the Eu3+ 4f6 configuration. The deviation and rms are 6.8 and 7.9 cm(-1), respectively for 38 levels and ten parameters. The experimental crystal field parameters are in good agreement with the ab-initio ones. Moreover, the relative intensities of the 5D0-->7F2,3,4 emissions are well reproduced by an 'ab-initio' calculation, except for three lines. The Er3+ ions introduced in LaZr2F11, microcrystals also lie in an unique crystallographic site. A total of 31 energy levels were recorded and the crystal field analysis led to 6.6 and 7.8 cm(-1) for the deviation and rms, respectively, for nine variable parameters taken into account. The experimental CF parameters for Er3+ and Eu3+ are very similar, which seems to show that the host lattice contracts around the smaller Er3+ ion. The informations given by both Eu3+ and Er3+ optical probes in LaZr2F11 are very consistent with the structure previously determined for the isotypic PrZr2F11 fluoride. PMID- 10794463 TI - Infrared E-type band profiles of acetonitrile in condensed media: orientational diffusion and free rotation. AB - Infrared spectra of liquid acetonitrile (CH3CN) and its solutions in CCl4, CS2, chloroform, dichloromethane, benzene-d6, acetone-d6 and dimethyl sulfoxide-d6 have been studied. E-type bands under investigation (v5 = 3009, v6 = 1448 and v7 = 1041 cm(-1)) were reproduced by the sum of two Cauchy-Gauss components, the narrower (n) and the broader (b) ones. The different temperature behaviour of the components has been found: the integrated intensities of the narrower components, In, decrease with the temperature, while the intensities of the broader ones, Ib, increase. The narrower components of the bands were explained within the framework of the orientational diffusion mechanism. The broader components of v6 and v7 were attributed to the unresolved gas-like vibration-rotational absorption of the molecules. The enthalpy difference between the molecules absorbing via these two different mechanisms was determined from the dependence of ln(In/Ib) upon T(-1) :deltaH0 = 1.26+/-0.15 kcal mol(-1). The broader component of v5 is assumed to be mainly due to interactions of C-H stretching vibrations with single particle and collective motions of molecular dipoles. The narrower components' widths were used for evaluating the spinning diffusion constant of CH3CN. The absorption in C-H stretching region was found to be strongly affected by solvent. These effects were explained within the framework of hydrogen bond formation between the CH3-group of acetonitrile and H-bond acceptor groups of the solvent molecules. PMID- 10794464 TI - Raman microscopic study at 300 and 77 K of some pegmatite minerals from the Iveland-Evje area, Aust-Agder, Southern Norway. AB - The Raman spectra at 300 and 77 K of beryl, columbite-tantalite and topaz single crystals from pegmatites in the Iveland-Evje area are described in detail. The beryl is shown to contain mainly water type I and less of type II in its channels, while CO2 is only a very minor channel constituent. Cooling to 77 K results in minor shifts towards higher wavenumbers for most bands. The Si-O vibrations at 1009 and 1066 cm(-1) show a doubling to four bands at 1015, 1072, 1087 and 1149 cm(-1) due to structural rearrangements in the hexameric rings forming the channels in the beryl crystal structure. In addition a new band becomes visible around 1155 cm(-1). The tantalite could not be analysed in detail due to strong fluorescence. The Raman spectrum of the yellow topaz from Solas is comparable to that of the colourless topaz from Topaz Mountain, Thomas Range, Utah. Upon cooling to 77 K, two OH-stretching bands become visible around 3644 and 3655 cm(-1), which were not observed at room temperature. PMID- 10794465 TI - Direct observation of intramolecular vibrational energy redistribution under white light excitation. II. Fluorescence spectra of fluorobenzene derivatives by controlled electron impact. AB - The low-resolution (resolution: 0.6 nm) fluorescence spectra of fluorobenzene derivatives, o-, m- and p-fluorotoluene (o-, m- and p-FT) were observed to investigate intramolecular vibrational energy redistribution (IVR) under controlled electron impact. The o- and m-FT spectra mainly consisted of unstructured emission from optically inactive states (bath modes) populated through IVR. The p-FT spectrum consisted of structured emission from optically active states and unstructured emission. The high-resolution (resolution: 0.15 nm) fluorescence spectrum of p-FT was measured to estimate the fraction of the structured emission intensity to the total emission intensity. The IVR rate of p FT under electron impact excitation was faster than that under laser excitation. The fraction did not depend on the incident energy of electrons from 20 to 200 eV, and thus the IVR acceleration is not attributable to breakdown of the Born approximation. PMID- 10794466 TI - Infrared intensity and morphology of l-monolaurin-water systems. AB - The infrared spectra for some metastable states in 1-monolaurin water systems were observed at room temperature, where the relative intensity of bands due to paraffin chains changed considerably, especially in the CH2 rockings, which disappear in some cases. It is considered that the spectral changes result from the morphology change on going from the crystal to the liquid crystal, smectic B phase, so-called gel phase, which consists of the lipid bilayers with ordered paraffin chains alternating with water layers. The model for explaining the intensity change is proposed on the basis of the interaction among oscillating dipoles. PMID- 10794467 TI - X-ray photoelectron spectra of complexes with 1-(D-3-mercapto-2-methylpropionyl) L-proline and Ni(II), Cd(II) and Cu(I): synthesis and LAXS study of Cu(I) derivative. AB - Metal complexes of general formula Na2M(CAP)2xH2O (with M = Cd(II) or Ni(II), x = 7 and 4, respectively, CAP = 1-(D-3-mercapto-2-methylpropionyl)-L-proline) and NaCuCAPx3H2O have been synthesized as amorphous compounds and studied by means of X-ray photoelectron spectroscopy (XPS). Cu(I) derivative has been studied by IR, XPS and large-angle X-ray scattering (LAXS). IR data and the chemical shift of core level signals suggest that CAP is bonded to the metal via the sulphur atom and the carbonylic oxygen. LAXS data confirm this finding and are consistent with a tetrahedral configuration around the copper ion. The CAP molecule is bonded through the sulphur and the carbonylic oxygen and two water molecules complete the coordination around the metal. The sodium ion exhibits a tetrahedral configuration and interacts with the carboxylic group and two water molecules. One of these is bridging bonded between copper and sodium. No metal-nitrogen bonds are present. PMID- 10794468 TI - The identification of vicinally substituted cyclohexane isomers in their mixtures by 1H and 13C NMR spectroscopy. AB - The radical addition reactions of organobromine compounds, XBr (X = CH2COOMe, PhCH2, CHBr2 and CCl3) with cyclohexene afforded mixtures of cis/trans isomer pairs of 1-X-2-Br-cyclohexanes. In addition to benzyl benzoyloxy derivatives are formed also, when benzoyl peroxide is used as an initiator. Owing to the great difficulties in separating these cis/trans isomer pairs, they are identified directly in their mixtures by NMR spectroscopy. In addition to one-dimensional (ID) 1H, proton decoupled 13C and DEPT-135, also two-dimensional (2D) 13C-13C INADEQUATE as well as 1H-13C HMQC experiments have been used in assigning the signals of each compound in their mixtures. The identification of each isomer was based on comparison of experimental 3J(H,H) coupling constants with theoretical ones based on the well-known Karplus type relationship. The more stable conformation for each isomer was estimated using the semiempirical AM1 molecular orbital method. The calculations support the isomer pair elucidations. PMID- 10794469 TI - Fourier transform Raman and infrared spectra and normal coordinate analysis of organo-arsenic(III), -antimony(III) and -bismuth(III) thiolates. AB - The FT-Raman and infrared spectra of (PhS)BiPh2, (PhS)2BiPh, (PhS)3Sb, (PhS)3Sb, (PhS)3As and (PhSe)BiPh2 were measured in the range 3600-100 cm(-1). A normal coordinate analysis was performed for all substances in terms of the calculation and discussion of the force constants which are dependent on the element-sulphur and element-selenium group using a modified valence force field. Furthermore, for all compounds 1H-NMR, 13C-NMR and MS data were also given. The investigation of the microbiological activity of some substances against Escherichia coli was an additional aspect because of their strong bactericidal and fungicidal effects. PMID- 10794470 TI - In situ infrared spectroscopic analysis of the adsorption of aromatic carboxylic acids to TiO2, ZrO2, Al2O3, and Ta2O5 from aqueous solutions. AB - In situ infrared spectroscopy has been used to investigate the adsorption of a range of simple aromatic carboxylic acids from aqueous solution to metal oxides. Thin films of TiO2, ZrO2, Al2O3 and Ta2O5 were prepared by evaporation of aqueous sols on single reflection ZnSe prisms. Benzoic acid adsorbed very strongly to ZrO2, in a bridging bidentate fashion, but showed only weak adsorption to TiO2 and Ta2O5. Substituted aromatic carboxylic acids; salicylic, phthalic and thiosalicylic, were found to adsorb to each metal oxide. Salicylic and phthalic acids adsorbed to the metal oxides via bidentate interactions, involving coordination through both carboxylate and substituent groups. Thiosalicylic acid adsorbed to the metal oxides as a bridging bidentate carboxylate with no coordination through the thiol substituent group. PMID- 10794471 TI - Density functional theory study on fundamental vibrational spectra of disilyl iodide and its isotopomer. AB - Density function theory calculations using B3' exchange functional and LYP' correlation functionals (B3LYP) with the 3-21G** basis set were carried out to study the molecular structure and fundamental vibrational frequencies of Si2H5Br, Si2H5I and their isotopomers. One scale factor used to scale Si-H(D) force constants for Si2H5Br is transferred from Si2H5Br to Si2H5I. Other scale factors of force constants for Si2H5I and Si2D5I are determined by a least-squares fitting to experimental vibrational frequencies. The predictions of fundamental vibrational frequencies for Si2H5I and its isotopomer are given. The scaled DFT force fields using B3LYP/3-21G** reproduced the observed fundamental vibrational frequencies of Si2H5I and its deuterium isotopomer with a mean absolute deviation of 6 cm(-1). Reassignment of some fundamental vibrational modes of Si2H5I and its isotopomer is discussed. PMID- 10794472 TI - Vibrational coupling in oxo-centred trinuclear clusters: oxygen-16/18 isotopic substitution studies of [Fe3III(O)(O2CC(CH3)3)6(py)3][FeCl4] and [Fe2(III)FeII(O)(O2CC(CH3)3)6(py)3]. AB - IR spectra are reported for the compounds [Fe3O(O2CC(CH3)3)6(py)3][FeCl4] and [Fe3O(O2CC(CH3)3)6(py)3]. Using isotopic substitution at the central oxygen atom, the assignments of the in-plane and out-of-plane vibrations of this atom are confirmed, and coupling is demonstrated between the in-plane modes v(as)(Fe3O) and carboxylate deformation modes rho(r)(C-CO2). PMID- 10794473 TI - Spectroscopic studies of copper(II) and iron(II) complexes of adriamycin. AB - The complexes of adriamycin (ADM) with Cu(II) and Fe(II) have been studied by visible absorption, circular dichroism (CD) and fluorescence spectra, respectively. In Tris buffer at pH 7.0, either metal ions forms a single species with adriamycin: Cu(ADM)2 or Fe(ADM)3. Interaction of these two complexes with various biological molecules has been examined. It is shown that some amino acids, glutathione and albumin are able to remove the Cu(II) ion from Cu(II)-ADM complex, releasing the free drug. However, Fe(II)-ADM keeps in an undissociated form under the same conditions. The possibility of Fe(II) ADM as a new alternative drug has been discussed. PMID- 10794474 TI - Theoretical and experimental study of the vibrational spectrum of N-acetyl-L alanine. AB - The energies, vibrational frequencies and IR intensities of cis- and trans-N acetyl-L-alanine (NAAL) are computed using the density functional theory (B3LYP) combined with the 6-311G(d, p) basis set. The trans conformer is characterized by an intramolecular NH ... O hydrogen bond leading to the formation of a five membered ring and is by 23 kJ mol(-1) more stable than the cis conformer. The difference between the vibrational frequencies and IR intensities computed for the two conformers is discussed. The IR spectra at different temperatures and the Raman spectra of solid NAAL and its deuterated counterpart are investigated and discussed. The frequencies of the v(OH) vibration and the isotopic ratio suggest the formation of short OH ... O hydrogen bonds in the solid state. The NH group seems also to be involved in a weak hydrogen bond. PMID- 10794475 TI - NH-S hydrogen bonding in zinc enzyme model complex with S2N2 binding set studied by normal coordinate analysis of vibrational spectra. AB - In order to provide theoretical evidence for the existence and effect of NH-S hydrogen bonding in the zinc enzyme model complex [Zn(S-2-C6H4NHCOC6H5)2(1 MeIm)2] (1-MeIm = 1-methylimidazole) in addition to the spectroscopic and crystallographic investigations, normal coordinate analysis (NCA) was carried out using a modified Urey-Bradley force field. The vibrational frequencies of the complex with and without the NH-S hydrogen bonding as well as the corresponding internal coordinates were obtained. The good agreement found between the calculated and observed frequencies in the presence of the NH-S hydrogen bond, supports the reliability of the analysis. The stretching force constant of the NH S hydrogen bond obtained from the calculation is 0.18 mdyne per A. The calculation shows that the N-H bond is weakened by formation of the NH-S hydrogen bond. The results are indicative of the existence of the NH-S hydrogen bond in the complex. PMID- 10794476 TI - Fluorescence properties and stability of dioxygen--binding functional units from the Rapana thomasiana hemocyanin subunit RHSS2. AB - Molecular aggregates of the Rapana thomasiana hemocyanin are composed of two structural subunits, RHSS1 and RHSS2, each of which contains eight functional units (FUs) reversibly binding dioxygen. Multiunit fragments and individual 50-60 kDa FUs from RHSS2 were isolated and characterized by electron and fluorescence spectroscopy. The units have similar fluorescence parameters demonstrating that the tryptophyl side chains are located in the hydrophobic core of the globular folded regions. The copper-dioxygen system at the binuclear active site stabilizes considerably the native protein structure and quenches the indole emission. The removal of this system decreased the 'melting points' drastically Tm by 13-20 degrees C and increased 2-4 times the fluorescence quantum yields. The individual FUs differ considerably in their thermostability. The activation energy for the thermal deactivation of the excited tryptophyl residues of the apo FUs is lower compared to that of the whole molluscan apo-Hcs. PMID- 10794477 TI - A prospective study of plasma insulin-like growth factor-1 and binding protein-3 and risk of colorectal neoplasia in women. AB - Insulin-like growth factor-1 (IGF-1) is an important mitogen, and IGF binding protein-3 (IGFBP-3) has opposing effects. Acromegalics, who have abnormally elevated levels of IGF-1, are at increased risk of colorectal tumors. Recent studies have found that IGF-1 levels correlate with risk of prostate cancer and colorectal cancer in men, premenopausal breast cancer in women, and lung cancer in men and women. We examined whether prediagnostic plasma levels of IGF-1 and IGFBP-3 influence risk of colorectal cancer and adenoma in women. From 1989 to 1990, a total of 32,826 women from the Nurses' Health Study provided blood specimens that were archived in liquid nitrogen. During 6 years of follow-up from 1989 to 1994, we documented 79 new cases of colorectal cancer, 90 cases of intermediate/late-stage adenoma (> or =1 cm or tubulovillous/villous histology), and 107 cases of early-stage adenoma (<1 cm and tubular histology). After matching controls (2:1 for cancers and 1:1 for adenomas) to cases by age, month of blood draw, fasting status, and indication for endoscopy (for adenoma controls), plasma IGF-1 and IGFBP-3 levels were measured. Controlling for IGFBP-3 level, relative to women in the low tertile of IGF-1, those in the high tertile were at elevated risk of intermediate/late-stage colorectal neoplasia adenoma [multivariate relative risk (RR), 2.78; 95% confidence interval (CI), 0.76-9.76] and cancer (RR, 2.18; 95% CI, 0.94-5.08). Controlling for IGF-1 level, relative to women in the low tertile of IGFBP-3, women in the high tertile of IGFBP-3 were at lower risk of intermediate/late-stage colorectal adenoma (RR, 0.28; 95% CI, 0.09-0.85) and cancer (RR, 0.28; 95% CI, 0.10-0.83). Neither IGF-1 nor IGFBP-3 had any appreciable relation with early-stage adenoma. These analyses indicate that high levels of circulating IGF-1 and particularly low levels of IGFBP-3 are associated independently with an elevated risk of large or tubulovillous/villous colorectal adenoma and cancer. PMID- 10794478 TI - Effect of subacute ibuprofen dosing on rectal mucosal prostaglandin E2 levels in healthy subjects with a history of resected polyps. AB - Nonsteroidal antiinflammatory drugs are among the most promising chemopreventive agents for colorectal cancer. Although the mechanism by which nonsteroidal antiinflammatory drugs exert such effects remains to be further characterized, their best known pharmacological effect is inhibition of prostaglandin synthetase, which leads to decreases in tissue prostaglandin levels. We conducted a randomized, double-blind, controlled study to examine the effect of daily ibuprofen treatment on the rectal mucosal prostaglandin E2 (PGE2) levels in healthy subjects with a history of resected polyps. Study participants (n = 27) completed a 2-week run-in period and were then randomized to take a single, daily dose of ibuprofen (300 or 600 mg) or of a placebo for 4 weeks. Rectal biopsy specimens were taken before and after the run-in period and at 2 and 4 weeks after the ibuprofen/placebo treatment. Notably large between- and within-subject variability in the rectal mucosal PGE2 content was seen. The changes in PGE2 levels after ibuprofen/placebo treatment correlated with the baseline PGE2 content. After adjustment of the baseline values, 2 weeks of 300 mg/day of ibuprofen treatment resulted in significantly more suppression of PGE2 levels than that observed after the placebo treatment (55% versus 22% suppression from baseline; P = 0.033). Although other ibuprofen treatment schedules and doses appeared to result in suppression in the PGE2 levels, the suppression was not statistically significant because of the large variability in this measurement. Because lower doses are associated with fewer adverse effects, a dose of 300 mg of ibuprofen/day should be considered for future Phase II chemoprevention studies. Stratifying study participants, based on their baseline PGE2 levels and inclusion of a larger number of study subjects, are recommended for future trials where the rectal mucosal PGE2 level is to be used as a surrogate end point biomarker. PMID- 10794479 TI - A prospective cohort study on antioxidant and folate intake and male lung cancer risk. AB - Many studies have reported inverse associations between vegetable and fruit consumption and lung cancer risk. The aim of the present study was to elucidate the role of several antioxidants and folate in this relationship. In the Netherlands Cohort Study on Diet and Cancer, 58,279 men of ages 55-69 years at baseline in 1986 returned a questionnaire including a 150-item food frequency questionnaire. After 6.3 years of follow-up, 939 male lung cancer cases were registered. A new Dutch carotenoid database was used to estimate intake of alpha carotene, beta-carotene, lutein + zeaxanthin, beta-cryptoxanthin, and lycopene, completed with the antioxidant vitamins C and E and folate. Using case-cohort analysis, rate ratios were calculated, adjusted for age, smoking, educational level, and family history of lung cancer. Protective effects on lung cancer incidence were found for lutein + zeaxanthin, beta-cryptoxanthin, folate, and vitamin C. Other carotenoids (alpha-carotene, beta-carotene, and lycopene) and vitamin E did not show significant associations. After adjustment for vitamin C, only folate remained inversely associated, and after adjustment for folate, only beta-cryptoxanthin and vitamin C remained significantly associated. Inverse associations were strongest among current smokers and weaker for former smokers at baseline. Inverse associations with carotenes, lutein + zeaxanthin, and beta cryptoxanthin seemed to be limited to small cell and squamous cell carcinomas. Only folate and vitamin C intake appeared to be inversely related to small cell and squamous cell carcinomas and adenocarcinomas. Folate, vitamin C, and beta cryptoxanthin might be better protective agents against lung cancer in smokers than alpha-carotene, beta-carotene, lutein + zeaxanthin, and lycopene. PMID- 10794480 TI - Applicability of induced sputum for molecular dosimetry of exposure to inhalatory carcinogens: 32P-postlabeling of lipophilic DNA adducts in smokers and nonsmokers. AB - The lung is a major target organ for smoking-associated cancer. We examined the applicability of induced sputum for molecular dosimetry of exposure to tobacco smoke-related carcinogens. Sputum induction was performed by inhalation of 4.5% saline delivered from an ultrasonic nebulizer for a period of up to 21 min in a group of smoking (n = 20) and nonsmoking (n = 24) healthy individuals. Samples were analyzed for total and differential cell counts and cell viability. Subsequently, DNA contents of the samples were isolated, and measurement of lipophilic DNA adducts was done by the 32P-postlabeling assay using nuclease P1 (NP1) and butanol enrichment methods. All subjects tolerated the induction procedure without experiencing any troublesome symptoms, and 90% of smokers (18 of 20) and 88% of nonsmokers (21 of 24) succeeded in producing sufficient amounts of sputum. Total cell counts and percentages of viable cells in smokers were higher than those in nonsmokers (6.7+/-6.0 versus 4.7+/-6.0 x 10(6), P = 0.40 and 80+/-15 versus 63+/-17, P = 0.01, respectively). In cell differentials, smokers had lower percentages of bronchoalveolar macrophages and higher percentages of neutrophils (69+/-24 versus 92+/-5, P = 0.002 and 26+/-26 versus 4+/-4, P = 0.008, respectively). Using the NP1 digestion method, all smokers and only one nonsmoker showed a diagonal radioactive zone in their adduct maps; adduct levels in smokers were higher than those in nonsmokers (3.1+/-1.4 versus 0.6+/-0.8/10(8) nucleotides; P = 0.0007), and also, adduct levels were significantly related to smoking indices. Applying the butanol extraction method, however, only half of the smokers and three nonsmokers showed the diagonal radioactive zone in their adduct maps; adduct levels in smokers were higher than those in nonsmokers (4.6+/ 3.7 versus 1.0+/-1.9/10(8) nucleotides; P = 0.02), and the levels of adducts were significantly related to the smoking indices. There was a correlation between the levels of adducts determined by the two enrichment methods (r = 0.7; P = 0.02). Paired comparison showed no differences between the levels of adducts measured by the two methods (P = 0.55). We conclude that induced sputum can serve for molecular dosimetry of inhalatory exposure to carcinogens and that the NP1 version of the 32P-postlabeling assay is a choice of preference for studying smoking-induced DNA adducts in the lower respiratory tract. PMID- 10794481 TI - Biomonitoring of exposure to polycyclic aromatic hydrocarbons of nonoccupationally exposed persons. AB - In a field study with 69 subjects, we investigated the influence of smoking, exposure to environmental tobacco smoke (ETS), diet, and location of residence on biomarkers for polycyclic aromatic hydrocarbons (PAH), including urinary excretion of 1-hydroxypyrene and benzo[a]pyrene (BaP) adducts of hemoglobin and albumin. The self-reported smoking status and the extent of ETS exposure were verified by urinary cotinine measurements. ETS exposure was quantified by nicotine and 3-ethenylpyridine measurements on personal samplers worn by the nonsmokers over 5 or 7 days before blood and urine samples were collected. Smokers (n = 27), on average, excreted 0.346 microg/24 h 1-hydroxypyrene, whereas the corresponding value for nonsmokers (n = 42) was 0.157 microg/24 h. Average BaP adduct levels with hemoglobin and albumin were 0.105 fmol/mg and 0.042 fmol/mg, respectively, for smokers, and 0.068 fmol/mg and 0.020 fmol/mg, respectively, for nonsmokers. The differences, except for the hemoglobin adducts, were statistically significant. Of the 42 nonsmokers, 19 were classified as passive smokers. There was no significant difference in the PAH biomarkers between nonsmokers exposed to ETS and those not or rarely exposed to ETS. Total dietary BaP intake, as calculated from questionnaire data, did not correlate with any of the PAH biomarkers (r < 0.1). Subjects living in the suburbs tended to have higher BaP-protein adduct levels than subjects living in the city. Our findings suggest that diet and smoking are major sources for PAH exposure of persons not occupationally exposed to PAH, whereas the influence of ETS exposure is negligible. The lack of correlation between the dietary PAH intake and the PAH biomarkers may be due to the inaccuracy of the estimate for the dietary PAH intake. PMID- 10794482 TI - Spiritual faith and genetic testing decisions among high-risk breast cancer probands. AB - Despite widespread access to genetic testing for the BRCA1 and BRCA2 breast cancer susceptibility genes, little is known about rates or predictors of test use among individuals from newly ascertained high-risk families who have self referred for genetic counseling/testing. The objective of this study was to examine rates of test use within this population. In addition, we sought to determine whether spiritual faith and psychological factors influenced testing decisions. Participants were 290 women with familial breast cancer. All were offered genetic counseling and testing for alterations in the BRCA1 and BRCA2 genes. Baseline levels of spiritual faith, cancer-specific distress, perceived risk, and demographic factors were examined to identify independent predictors of whether participants received versus declined testing. The final logistic model revealed statistically significant main effects for spiritual faith [odds ratio (OR), 0.2; 95% confidence intervals (CIs), 0.1 and 0.5] and perceived ovarian cancer risk (OR, 2.4; 95% CIs, 1.3 and 4.7) and a statistically significant spiritual faith by perceived risk interaction effect. Among women who perceived themselves to be at low risk of developing breast cancer again, those with higher levels of spiritual faith were significantly less likely to be tested, compared with those with lower levels of faith (OR, 0.2; 95% CIs, 0.1 and 0.5). However, among women with high levels of perceived risk, rates of test use were high, regardless of levels of spiritual faith (OR, 1.2; 95% CIs, 0.4 and 3.0). These results highlight the role that spirituality may play in the decision-making process about genetic testing. PMID- 10794483 TI - TA repeat polymorphism of the 5alpha-reductase gene and breast cancer. AB - There is increasing evidence that androgens play a significant role in the development and progression of breast cancer. 5alpha-Reductase (SRD5A2) is an enzyme that is expressed in androgen-dependent tissues, and it catalyzes the reduction of testosterone to its more bioactive form, dihydrotestosterone, which then transactivates a number of genes. One of these genes encodes for prostate specific antigen (PSA), a favorable prognostic factor in breast cancer. The 3' untranslated region of the SRD5A2 gene contains either no TA repeats [(TA)0] or 9 [(TA)9] or 18 [(TA)18] repeats. Variations in the length of these dinucleotide repeats have been reported to influence the enzymatic activity of SRD5A2. In this study, we determined the TA genotypes in DNA from 141 well-characterized breast tumors and in DNA from whole blood of 70 women without cancer. The presence of TA genotypes was then associated with tumor cytosolic PSA concentrations and with clinicopathological variables, including disease-free survival and overall survival. Three genotypes, (TA)0 homozygote, (TA)0/(TA)9 heterozygote, and (TA)9 homozygote, were identified. No (TA)18 alleles were detected in any of the two patient groups. A statistically significant association between high PSA concentrations and (TA)0/(TA)9 or (TA)9 genotypes was observed (P = 0.004). (TA)0/(TA)9 or (TA)9 genotypes were found less frequently in patients at stage III or IV disease. TA genotypes were not associated with other clinicopathological variables by contingency table analysis. Patients with (TA)0/(TA)9 or (TA)9 repeats, when compared to those with genotypes homozygous for the (TA)0 allele, showed a significant reduction in the risk for relapse (P = 0.043). Long-term studies are needed to investigate the relevance of this polymorphism to breast cancer susceptibility. PMID- 10794484 TI - E2F-1: a proliferative marker of breast neoplasia. AB - E2F-1 is the best known ultimate transcription factor in the cyclin/cyclin dependent kinase/retinoblastoma gene pathway and is probably involved in carcinogenesis and tumor progression. Because E2F-1 can be detected in paraffin sections using immunohistochemical techniques, it could be a useful tumor/proliferation marker. We studied the expression of this gene product in 130 breast tissue specimens from 100 patients and compared it with the expression of Mib-1, the widely used prognostic/proliferative marker, to assess E2F-1 as a new marker of neoplastic proliferation. The percentage of E2F-1-positive cells increased from 1.9% in the normal breast (NB) to 6.3% in ductal carcinoma in situ (DCIS) and to 15.3% in invasive ductal carcinomas (IDC). In addition, higher grade tumors as well as advanced-stage disease correlated with higher expression of E2F-1. A similar tendency of Mib-1 expression was observed. There was a positive correlation between the E2F-1 and Mib-1 indices. In an in vitro experiment, we found that a similar difference in the expression of E2F-1 existed between a nontumorigenic breast cell line and two widely used breast carcinoma cell lines. The breast carcinoma cell lines T-47D and MCF-7 had more E2F-1 positive cells than the nontumorigenic cell line MCF-10F by immunohistochemistry and Western blot analysis. Because E2F-1 expression was significantly higher in IDC and DCIS than in NB, this study indicates that deregulation of E2F-1 may be involved in the development of breast IDC. In addition, E2F-1 expression could also be involved in tumor progression because the increased E2F-1 index correlated with the known prognostic predictors of breast cancer, such as histological grade, stage, metastasis status, estrogen receptor/progesterone receptor and Mib-1 expression. Thus, E2F-1 is a promising candidate to become a new prognostic/predictive marker of breast cancer. PMID- 10794485 TI - Reproducibility studies and interlaboratory concordance for androgen assays in female plasma. AB - We conducted studies to determine the magnitude and sources of variability in androgen assay results and to identify laboratories capable of performing such assays for large epidemiological studies. We studied androstanediol (ADIOL), androstanediol glucuronide (ADIOL G), androstenedione (ADION), androsterone glucuronide (ANDRO G), androsterone sulfate (ANDRO S), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEA S), dihydrotestosterone (DHT), and testosterone (TESTO). A single sample of plasma was obtained from five postmenopausal women, five premenopausal women in the midfollicular phase of the menstrual cycle, and five women in the midluteal phase, divided into aliquots, and stored at -70 degrees. Four sets of two coded aliquots from each woman were then sent to participating labs for analysis at monthly intervals over 4 months. Using the logarithm of assay measurements, we estimated the components of variance and three measures of reproducibility. The usual coefficient of variation is a function of the components that are under the control of the laboratory. The intraclass correlation between measurements for a given individual is the proportion of the total variability that is associated with individuals. The minimum detectable relative difference is important to evaluate study feasibility. Results suggest that a single sample of ADIOL G, DHEA, DHEA S, and ANDRO G (with two lab replicates per sample) can be used to discriminate reliably among women in a given menstrual phase or menopausal status. The results for DHT, TESTO, ADION, and ANDRO S are more problematic and suggest that the present measurement techniques should be used with care, especially with midluteal phase women. The results for ADIOL suggest that this assay is not yet ready for use in epidemiological studies. PMID- 10794486 TI - Urinary pharmacokinetics of the glucuronide and sulfate conjugates of genistein and daidzein. AB - Consumption of soybean-rich diets is thought to provide significant health benefits such as prevention of cancer, primarily because of the high contents of factors such as the isoflavones genistein and daidzein. Isoflavones circulate and are excreted into the urine mainly as glucuronide and sulfate conjugates. This study was conducted to determine the urinary pharmacokinetics of sulfate and glucuronide conjugates of genistein and daidzein. Twelve volunteers consumed a soy beverage providing 1 and 0.6 mg/kg body weight of genistein and daidzein equivalents, respectively. Urine was collected at various times during the 48 h after soy consumption and was digested with either glucuronidase or sulfatase, and the liberated aglycones were extracted and analyzed by liquid chromatography mass spectrometry. Urinary isoflavone sulfate levels were determined by two methods: (a) assessment of aglycone after sulfatase hydrolysis (measured); or (b) calculated by subtracting the aglycone + glucuronide levels from the total urinary isoflavone levels. The apparent terminal half-life for daidzein sulfate (3.9+/-0.5 h) that was determined from sulfatase-treated urine was 32% shorter (P < or = 0.02) than that of the calculated daidzein sulfate (5.7+/-0.08 h). A similar trend was obtained for genistein sulfate (4.5+/-0.7 versus 6.8+/-0.1 h). The apparent terminal half-lives for genistein and daidzein glucuronides were 6.0+/-0.4 and 3.8+/-0.4 h, respectively. These data suggest that the measured urinary isoflavone sulfate values provide a better understanding of the pharmacokinetics than the calculated values. Additional studies are needed to determine whether the apparent terminal half-lives can be attributed to elimination or absorption processes. PMID- 10794487 TI - Plasma xanthophyll carotenoids correlate inversely with indices of oxidative DNA damage and lipid peroxidation. AB - Post hoc analysis of data obtained from a study designed to modulate oxidative damage by dietary intervention revealed consistently strong inverse correlations between plasma xanthophyll carotenoids and oxidative damage indices. Thirty-seven women participated in a 14-day dietary intervention that increased mean vegetable and fruit (VF) consumption to approximately 12 servings/day. An additional 10 subjects participated in an intervention that limited VF consumption to less than four servings per day. 8-Hydroxy-2'-deoxyguanosine (8-OHdG) in DNA isolated from peripheral lymphocytes and 8-OHdG excreted in urine were measured as indices of oxidative DNA damage. Lipid peroxidation was assessed by measuring 8 epiprostaglandin F2alpha (8-EPG) in urine. Plasma levels of selected carotenoids were also determined, with the intention of using a-carotene as a biochemical index of VF consumption. Urinary 8-OHdG and 8-EPG were measured by ELISA, and plasma carotenoids were measured by high performance liquid chromatography. Lymphocyte 8-OHdG was measured by reverse phase high performance liquid chromatography with electrochemical detection. We observed that the structurally related xanthophyll carotenoids, lutein and beta-cryptoxanthin, which occur in dissimilar botanical families, were consistently inversely associated with these oxidative indices. Statistically significant inverse correlations were observed between plasma lutein and/or beta-cryptoxanthin levels and lymphocyte 8-OHdG and urinary 8-EPG. Moreover, an inverse correlation was observed between change in plasma xanthophylls and change in lymphocyte 8-OHdG concentration that occurred during the course of the study. These data lead us to hypothesize that lutein and beta-cryptoxanthin serve as markers for the antioxidant milieu provided by plants from which they are derived. Whether these carotenoids are directly responsible for the observed antioxidant phenomena merits further investigation. PMID- 10794488 TI - Lack of association between the C677T MTHFR polymorphism and colorectal hyperplastic polyps. AB - Colorectal hyperplastic polyps are benign lesions that share many risk factors with colorectal adenomas and cancers. Low folate intakes are associated with an increased risk of colon cancer. The enzyme 5,10-methylene-tetrahydrofolate reductase (MTHFR) may be linked to DNA methylation and nucleotide synthesis and thus play a role in the etiology of colorectal neoplasia. We investigated an association between the common MTHFR polymorphism (C677T) and colorectal hyperplastic polyps within the Minnesota Cancer Prevention Research Unit case control study. Cases (n = 200) were diagnosed with colonoscopically confirmed hyperplastic polyps; controls (n = 645) were derived from the same gastroenterology practice and were polyp-free at colonoscopy. Dietary intakes were estimated from a self-administered food-frequency questionnaire prior to colonoscopy. Multivariate adjusted odds ratios (ORs) and 95% confidence intervals for MTHFR status were 0.8 (0.6-1.2; CT versus CC wild-type) and 0.9 (0.5-1.6; TT versus CC). In subgroup analyses stratified on dietary intakes of folate, vitamin B12, vitamin B6, or methionine, those with the TT genotype and either low intakes of folate or vitamin B6 were at increased risk relative to those with normal or high vitamin intake. However, most 95% confidence intervals included 1.0, and no consistent trends were observed. In contrast to our findings on colorectal adenomas, increasing alcohol consumption was associated with an elevated risk of colorectal hyperplastic polyps, regardless of genotype. The MTHFR (C677T) variant genotype does not appear to be related to risk of colorectal hyperplastic polyps, and there is no convincing evidence that MTHFR shows a different relation to risk, dependent on dietary intakes of nutrients related to its pathway. PMID- 10794489 TI - p53 codon 72 polymorphism does not affect the risk of cervical cancer in patients from northern Italy. AB - A case-control study was performed to investigate the risk of cervical cancer associated with p53 polymorphism at codon 72, encoding either arginine or proline. It has been recently suggested that the arginine isoform increases the susceptibility to invasive cervical cancer; however, data remain controversial. The polymorphism was examined by both allele-specific PCR and RFLP analysis in 101 patients with primary cervical cancer and in 140 healthy women of the same age and from the same geographical area. The distribution of p53 genotypes in cervical cancer patients and in controls was not significantly different (P = 0.445), and homozygosity for arginine at residue 72 was not associated with an increased risk for cervical cancer (odds ratio, 0.81; 95% confidence interval, 0.47-1.42; P = 0.52). Similarly, different genotype distribution and increased risk were not observed when patients versus controls were analyzed according to human papillomavirus status and cancer histotype. Therefore, no evidence of association between homozygosity for p53 arginine and cervical cancer was found in our population sample. PMID- 10794491 TI - A case-control study of use of postmenopausal female hormone supplements in relation to the risk of large bowel cancer. AB - Postmenopausal female hormone use has been associated with a reduced risk of colon cancer. We assessed the relation of use of these supplements to the risk of large bowel cancer. The data were collected in a case-control study of large bowel cancer conducted in Massachusetts. Control subjects were matched to incident cases of carcinoma of the colon or rectum on age, gender, and town precinct. The analysis was restricted to women who experienced a natural menopause or had had a hysterectomy with or without removal of the ovaries (292 colon cancer cases and 112 rectal cancer cases and their matched controls). Use of female hormone supplements was associated with a decreased risk of colon cancer among recent users (odds ratio, 0.6; 95% confidence interval, 0.4-1.0) and long duration (5+ years) of use (odds ratio, 0.5; 95% confidence interval, 0.3 0.9). The association with long duration of use appeared to be independent of recency of use and screening practices and was apparent for late-stage cancer. Hormone supplement use was not associated with a reduced risk of rectal cancer. Our results add to the evidence for a decreased risk of colon cancer associated with use of female hormone supplements. PMID- 10794490 TI - The polymorphic exon 1 androgen receptor CAG repeat in men with a potential inherited predisposition to prostate cancer. AB - Recent studies have provided epidemiological evidence in support of a possible prostate cancer susceptibility locus on the X chromosome. The androgen receptor (AR) gene, located at Xq11-12, has been implicated as a risk factor for the development of prostate cancer. To examine the potential role of the AR locus in prostate cancer susceptibility, the AR CAG repeat length was measured in 270 Caucasian men with prostate cancer from 133 unrelated families. Each of these families has two or more confirmed cases of prostate cancer occurring in first- and/or second-degree relatives. No evidence for linkage of the AR gene to prostate cancer was observed. We tested for the previously reported association of short CAG alleles with prostate cancer using t tests, Pearson's chi2 tests, and logistic regression; analyses were subsequently repeated to incorporate only men with moderate- to high-grade prostate cancer. No association between AR CAG allele length and prostate cancer was detected when either a subset of unrelated patients or a subset of unrelated patients with moderate- to high-grade cancer was compared with a set of unrelated controls. We failed to detect an association between short AR CAG alleles and early age of prostate cancer diagnosis. Once specific hereditary prostate cancer genes have been identified, future studies can more carefully delineate the potential role of this AR polymorphism as a modifier locus in high-risk families. PMID- 10794492 TI - Glutathione S-transferase M1, T1, and P1 polymorphisms as risk factors for renal cell carcinoma: a case-control study. AB - Renal cell carcinoma (RCC) has known environmental risk factors, notably smoking, and enzymes that biotransform carcinogens have high levels of activity in the kidney. However, a possible role of polymorphisms in these enzymes in RCC etiology has received little study. We investigated glutathione S-transferase (GST) polymorphisms in a population-based case-control study of RCC. Subjects completed a structured interview, and DNA was isolated from pathological material or buccal cells for 130 cases, and from blood for 505 controls. Genotypes for GSTM1 and GSTT1 were determined by multiplex PCR, and for GSTP1 by oligonucleotide ligation assay. The frequency of GSTM1 null genotype was 50.0% in cases and 50.5% in controls, with an adjusted odds ratio (OR) of 1.0 [95% confidence interval (CI), 0.6-1.6]. For GSTP1, the frequencies of genotypes AA, AG, and GG representing the Ile104Val variant were: cases, 44.6%, 43.1%, and 12.3%; controls, 43.4%, 44.0%, and 12.6%; OR for AG and GG, 1.0 (95% CI, 0.6 1.6). An excess of the GSTT1 null genotype was observed in cases compared with controls, 28.6% versus 18.5% (OR, 1.9; 95% CI, 1.1-3.4). The association with GSTT1 was present among both smokers and nonsmokers, but was modified by body mass index, a recognized risk factor for RCC; among subjects in the lowest tertile of body mass index, the OR for GSTT1 null was 4.8 (95% CI, 1.8-13.0). The association between GSTT1 null and increased RCC risk in this population-based study suggests that activity of the GSTT1 enzyme protects against RCC. This contrasts with a recent report of reduced risk of RCC associated with GSTT1 null in a cohort of trichloroethene-exposed workers and suggests that specific chemical exposures alter the effect of GSTT1 on cancer risk. PMID- 10794493 TI - Reduced interleukin-1 responsiveness in immune system and central nervous system of inbred polydipsic (STR/N) mice. AB - Inbred polydipsic mice (STR/N strain) have primary polydipsia. The previous studies found abnormalities in the central nervous system (CNS), especially in the hypothalamus and circumventricular organ. As a part of pursuing to find the cause of the polydipsia, we investigated immunological characteristics of STR/N mice, using the ICR strain of mice as control. Their thymic subset cells showed that CD4+CD8+ double positive cells were increased, CD4+ single positive cells were decreased and CD5 expression was deficient, compared to ICR mice. T cell proliferative response and interleukin (IL)-2 production caused by IL-1beta stimulation were reduced in STR/N mice than those in the ICR mice. In in vivo studies the degree of thymic atrophy and the increases in serum level of ACTH and corticosterone induced by intraperitoneal IL-1beta injection were much less in STR/N mice than those in controls. Furthermore, adipsic response also induced by IL-1beta injection was greatly reduced compared to their control mice. All these results suggest that the responsiveness to IL-1 is impaired both in the immune system and the CNS of STR/N mice. PMID- 10794494 TI - Reduced food consumption increases water intake and modulates renal aquaporin-1 and -2 expression in autoimmune prone mice. AB - Aquaporin-1(AQP1) and AQP2 are members of the aquaporin family of cell membrane water channel transport proteins and have been implicated in the regulation of renal water excretion. We have previously shown that calorie restriction (CR) relative to ad libitum (AL) feeding extends lifespan and delays the onset of autoimmune kidney disease in lupus-prone (NZBxNZW)F1 (B/W) mice. To determine if AQP1 and/or AQP2 expression is influenced by CR, mice were fed an AL or CR (40% less food) diet until 4 (young) or 9 (old) months of age when mice were sacrificed. Kidneys were removed and the expression of AQP1 and AQP2 was determined at the protein and mRNA levels using western blotting and RT-PCR respectively. While age did not significantly increase AQP1 expression in the AL groups, CR did increase both the protein (1.4-fold) and mRNA (2.4-fold) levels. In old mice, AQP1 expression was higher (1.8-fold) in CR compared to the AL group while CR had no effect in young mice. In contrast, AQP2 showed an age related decrease (55%) in the AL groups and an increase in the protein (8.4-fold) and mRNA (1.7-fold) levels in the CR groups. Relative to AL, CR decreased AQP2 expression at the protein (90%) and mRNA (50%) levels in the young mice while an increase at the protein (2.9-fold) and mRNA (1.9-fold) levels was evident in the old mice. Interestingly, a significant increase in water intake per gram body weight was found in both young and old CR fed mice when compared to their AL counterparts which may contribute to the prevention of autoimmune disease with age and differences in longevity. These data show, for the first time, significant age and diet influences in renal AQP1 and AQP2 expression at both protein and mRNA levels in lupus-prone mice. PMID- 10794495 TI - Expressions of putative neurotransmitters and neuronal growth related genes in Merkel cell-neurite complexes of the rats. AB - Reverse transcription-polymerase chain reaction using 32P-labeled dCTP with specific primers for putative neurotransmitters related and neuronal growth related genes (GAP-43, NGF and BDNF) were used to search for evidence of such substances in the Merkel cell. Merkel cell samples were made from sinus hairs in the facial skin of rats. The relative amount of mRNA in a tissue sample concentrated in Merkel cells was compared semiquantitatively to that from nearby tissue without Merkel cells. mRNAs for VIP, tyrosine hydroxylase, substance P were found in higher concentration in Merkel cells than in control tissues. mRNA for genes encoding pro-enkephalin, GAP-43, CGRP, NGF and BDNF were detected in the Merkel cell samples at concentration statistically equivalent to those found in the control tissues. It was concluded that the relative concentration of mRNAs for VIP, tyrosine hydroxylase and substance P is consistent with the possibility that Merkel cell acts as a possible transduction element in mechanical excitation of sense organs in which Merkel cells are present. PMID- 10794497 TI - Infant formula ingestion is associated with the development of diabetes in the BB/Wor rat. AB - The association between early exposure to cow's milk products in infancy and risk for insulin dependent diabetes mellitus (IDDM) is controversial. We examined whether the ingestion of cow's milk-based infant formula altered the expression of the diabetic syndrome in the BB/Wor rat, an animal model of IDDM. Pregnant BB/Wor dams were obtained from the NIH contract colony at the University of Massachusetts and housed under semi-barrier conditions. Rat pups were intubated with 1 to 2 ml of commercially available cow's milk-based infant formula (Enfamil or Nutramigen) or sham intubated (controls) daily from day 12 to day 25 of life. Pups were weaned at day 25 and monitored for glucosuria daily through 120 days of life. All rats including dams consumed a milk-free rat chow and acidified water ad libitum throughout the study. The mean age of disease onset was 4 to 10 days earlier in Nutramigen-fed and Enfamil-fed rats relative to controls (84+/-3, 78+/ 2 and 88+/-4 days, respectively); the mean age of disease onset was significantly different between controls and Enfamil-fed animals (p<0.05). At 120 days, 60% (12/20) of control rats developed diabetes versus 100% of animals fed either type of infant formula prior to weaning (15/15:Enfamil-fed; 19/19:Nutramigen-fed) (p<0.05). These data indicate that direct, early ingestion of cow's milk-based formula was related to the expression of diabetes in the BB/Wor rat. PMID- 10794496 TI - Measurements of 1,2-diacylglycerol and ceramide in hearts subjected to ischemic preconditioning. AB - An accumulation of recent evidence suggests that the mechanism in ischemic preconditioning (IPC) may involve the activation of protein kinase C (PKC) regulatory pathway. In this study, we examined whether the content of 1,2 diacylglycerol (1,2-DAG) and ceramide, which are intracellular second messengers regulating PKC activity, change during IPC in isolated perfused rat hearts, and whether the observed change in 1,2-DAG is accompanied with alteration in its fatty acid composition. Hearts subjected to IPC, consisting of 5-min transient global ischemia followed by 5-min reperfusion, presented a significant functional recovery during subsequent 40-min reperfusion following 40-min global ischemia compared with non-preconditioned hearts. An increase in 1,2-DAG content was observed in hearts subjected to 5-min transient ischemia compared with non ischemic control hearts, however this was not seen in hearts harvested after 5 min reperfusion following 5-min ischemia. While fatty acid composition in 1,2-DAG was virtually unchanged in hearts subjected to 5-min ischemia, saturated 1,2-DAG decreased and monounsaturated/polyunsaturated 1,2-DAG increased in hearts reperfused for 5-min following 5-min ischemia compared with the non-ischemic control hearts. Ceramide mass did not change significantly, suggesting that the contribution of ceramide may be small in IPC. These data are in concert with the hypothesis that 1,2-DAG is a second messenger in IPC and the changes in fatty acid composition of 1,2-DAG may add new insight concerning signal transduction pathway in IPC. PMID- 10794498 TI - Contribution of circulating acetylcholine to sensory nerve conduction augmentation. AB - We previously discovered that sensory nerve action potential amplitudes increased during isometric muscle contraction and that this response could be blocked with tourniquet isolation of the contraction source. The hypothesis for this study was that a circulating factor was responsible for this effect. In this prospective study, baseline and post intravenous injection of serial sural nerve action potential recordings were made in the leg of 8 rabbits. The sequence of the injections was randomized: 1) normal saline placebo, 2) 0.01 mg/kg acetylcholine (ACh) 3) 200 mg/kg Na acetate, 4) 260 mg/kg Na lactate, and 5) 20 mg/kg choline. Results showed there was a 3.8 microV increase in the sural nerve response 6 min after ACh injection compared to baseline at rest (p = .01, power = .9, analysis of variance (ANOVA), repeated measures). There were no significant changes in the amplitudes of the sural nerve after injection of the remaining agents or placebo (p = .33 to .81, ANOVA, repeated measures). In conclusion, circulating ACh is the only agent tested thus far that appears to be responsible for this effect. In addition, the amplitude and temporal curve of this response is similar to that seen after exercise in human subjects. The clinical importance of this study is that ACh plays a role in this newly discovered sensory regulatory mechanism controlled by the motor system. PMID- 10794499 TI - L-arginine supplementation in hypercholesterolemic rabbits normalizes leukocyte adhesion to non-endothelial matrix. AB - L-arginine slows the development of atheromatous lesions, improves endothelium dependent relaxation, and reduces the vascular superoxide anion production in hypercholesterolemic rabbits. These beneficial effects have been attributed to L arginine-dependent formation of nitric oxide within the endothelial layer; a direct effect of L-arginine on other cells, however, has not been investigated. We hypothesised that in hypercholesterolemia L-arginine also specifically acts via a direct inhibitory effect on leukocytes, without affecting endothelial cells. The action of L-arginine was compared to vitamin E and the HMG CoA reductase inhibitor lovastatin which are known to attenuate progression of atherosclerosis. Rabbits were fed cholesterol enriched diet and from week five on lovastatin (10 mg/day), vitamin E (300 mg/d) or L-arginine (2% in drinking water) were given. After 16 weeks, blood cholesterol concentration was determined and leukocyte adhesion to cotton wool was measured. In order to exclude any endothelium-mediated effects an adhesion assay to endothelial cells was avoided. Cholesterol-enriched diet increased plasma cholesterol concentration (19+/-3 vs. 1427+/-117 mg/dl). Cholesterol levels were not affected by L-arginine (1344+/-163 mg/dl) or vitamine E (1312+/-243 mg/dl). Lovastatin treatment reduced cholesterol concentration by 35% as compared to the cholesterol group (899+/-51, p<0.05 vs. cholesterol). Cholesterol diet significantly increased leukocyte adhesion to cotton wool (16+/-3% vs 27+/-4%, p<0.05). Lovastatin or vitamine E had no effect on leukocyte adhesion (31+/-4%, 39+/-5), whereas L-arginine completely normalized adhesion (8.8+/-3%). CONCLUSION: Rabbits fed high cholesterol diet have increased leukocyte adhesion, which is not affected by lovastatin or vitamine E treatment, but prevented by L-arginine supplementation. A direct inhibitory effect of L arginine on leukocyte adhesion may contribute to the beneficial effects observed with this substance. PMID- 10794500 TI - Blockage of amyloid beta peptide-induced cytosolic free calcium by fullerenol-1, carboxylate C60 in PC12 cells. AB - Amyloid beta protein (Abeta) alters signal transduction systems, including increases in the cytosolic free calcium ([Ca2+]i) response which have pathophysiological significance in Alzheimer's disease (AD). The purposes of this study were to elucidate the mechanism involved in Abeta's effect on the Ca2+ signal and to evaluate the effect of fullerenol-1, a water-soluble hydroxyl and superoxide radical scavenger, on the Abeta-induced Ca2+ response. Both Abeta and bradykinin (BK) dose-dependently elevated [Ca2+]i in PC12 cells. Fullerenol-1, at a concentration range between 100 nM and 1 microM, dose-dependently reduced the Abeta-induced [Ca2+]i response, but did not alter the subsequent BK-mediated process. Thapsigargin, an inhibitor of Ca2+-ATPase, released Ca2+ from the internal store and diminished the BK-mediated calcium spike but did not affect the Abeta-induced Ca2+ response. In the absence of extracellular calcium, the Abeta-induced, but not BK-induced, calcium spike was completely abolished. The Ca induced by Abeta did not enter through the voltage-dependent calcium channels or ligand gated calcium channels, because the peak of Abeta-evoked Ca2+ was not significantly altered by various Ca2+ channel blockers or a NMDA receptor antagonist MK801. In addition, neither cholera toxin nor pertussis toxin altered the Abeta-induced Ca response. The results demonstrated that Abeta-stimulated [Ca2+]i increase is due to Ca influx from an extracellular source rather than from the intracellular store. Alteration of the membrane lipid structure and permeability by free radicals generated by Abeta may be a major cause of Ca influx. Furthermore, fullerenol-1, a novel antioxidant, may provide therapeutic benefits in neurodegenerative diseases such as AD. PMID- 10794501 TI - Alpha-lactorphin lowers blood pressure measured by radiotelemetry in normotensive and spontaneously hypertensive rats. AB - Cardiovascular effects of subcutaneous administration of synthetic alpha lactorphin, a tetrapeptide (Tyr-Gly-Leu-Phe) originally derived from milk alpha lactalbumin, were studied in conscious spontaneously hypertensive rats (SHR) and in normotensive Wistar Kyoto rats (WKY) with continuous radiotelemetric monitoring. Alpha-lactorphin dose-dependently lowered blood pressure (BP) without affecting heart rate in SHR and WKY. The lowest dose which reduced BP was 10 microg/kg, and the maximal reductions in systolic and diastolic BP (by 23+/-4 and 17+/-4 mm Hg, respectively) were observed at 100 microg/kg dose in SHR. No further reductions were obtained at a higher dose of 1 mg/kg. There were no significant differences in the BP responses to alpha-lactorphin between SHR and WKY. Naloxone (1 and 3 mg/kg s.c.), a specific opioid receptor antagonist, abolished the alpha-lactorphin-induced reduction in BP and reversed it into a pressor response, which provides evidence for an involvement of opioid receptors in the depressor action of the tetrapeptide. PMID- 10794502 TI - Cellular proliferation and telomerase activity in CHRF-288-11 cells. AB - Telomerase activity is detected in many immortalized cell lines. Recent studies suggest that terminal differentiation of some of these cell lines is associated with a reduction in telomerase activity. However, the question remains whether the reduction in telomerase activity results from terminal differentiation or from cessation of cellular proliferation. This was explored in the megakaryocytic cell line CHRF-288-11. Cells were treated with phorbol 12-myristate 13-acetate (PMA), which induces terminal differentiation of CHRF-288-11 cells, EGTA, serum depletion, and okadaic acid. All treatments resulted in cessation of proliferation. Except for okadaic acid, these treatments also induced inhibition of telomerase within 7 days. Restoring the original growth conditions of cells treated with PMA, EGTA and serum depletion resulted in the reversal of telomerase inhibition and an acceleration of proliferation. Apparent inhibition of telomerase was observed to follow the cessation of proliferation, whereas enhanced telomerase activity was noted to precede acceleration in proliferation. Thus, telomerase activity usually reflects the proliferative status rather than the differentiated status of CHRF-288-11 cells. PMID- 10794503 TI - Induction of apoptosis and inhibition of DNA topoisomerase-I in K-562 cells by a marine microalgal polysaccharide. AB - We have previously purified an extracellular polysaccharide, D-galactan sulfate associated with L(+)-lactic acid, produced from a marine microalga Dinoflagellate Gymnodinium sp. A3 (GA3). The GA3 polysaccharide, irrespective of presence or absence of lactic acid, exhibited significant cytotoxicity, which is based on an induction of apoptotic cell death, toward human myeloid leukemia K562 cells. Furthermore, we found that the GA3 polysaccharide with or without lactic acid possesses an inhibitory effect on topoisomerase-I (topo-I). The potent cytotoxic effect of GA3 polysaccharide may result from its inhibitory effect on topo-I, because the topo-I inhibition is known to trigger apoptotic cell death. PMID- 10794504 TI - Differential loss in function of angiotensin II receptor subtypes during tissue storage. AB - In vitro receptor autoradiography was performed on rat brain and kidney sections stored frozen at -20 degrees C for extended time periods (17, 40, 64, 121, 183, 251, and 333 days). The results indicate that prolonged tissue storage has a differential effect upon 125I sar1ile8 angiotensin II binding to AT1 and AT2 receptor sites. Binding at AT1 receptor rich tissues studied (renal medulla, renal cortex, anterior pituitary, ventral hippocampus, spinal trigeminal nucleus, and nucleus of the solitary tract) shows a first order exponential decay pattern. The logarithmic linear regression slope (log(e) specific binding versus time), is significantly different from zero (p<0.05) in all AT1 rich tissues except for nucleus of the solitary tract (p=0.086). There is no detected loss of 125I sar1ile8 angiotensin II binding at the AT2 prominent regions in the superior colliculus, medial geniculate nucleus, and the inferior olivary nucleus. The half lives of AT1 receptors are highly variable, ranging from 36 days in the anterior pituitary to 442 days in the nucleus of the solitary tract, and this might be related to variable stability of AT1A and AT1B receptors. These observations should be taken into account when assessing and comparing AT1 and AT2 receptor subtype densities. PMID- 10794505 TI - Possible requirement of phosphonate moiety for efonidipine effects on the sino atrial node action potential. AB - The effects of efonidipine, a 1,4-dihydropyridine phosphonate, and structurally related compounds on rabbit sino-atrial node action potential were examined with microelectrodes. 3NIC5NZ has a phosphonate moiety identical to that of efonidipine at the C5 position of the dihydropyridine ring and a side chain identical to nicardipine at C3, while 3NZ5NIC has C5 and C3 side chains identical to nicardipine and efonidipine, respectively. All four compounds decreased the slope and prolonged the early and late phases of pacemaker depolarization. The selectivity for the late phase against the early phase was in the order of efonidipine > 3NIC5NZ >> nicardipine > 3NZ5NIC. Thus, the phosphonate moiety at C5 position of the may be important for the characteristic prolongation of the late phase pacemaker depolarization by efonidipine. PMID- 10794506 TI - In vitro stability of polyether and polycarbonate urethanes. AB - The in vitro structural stability of poly-ether-urethanes (PEUs) and poly carbonate-urethanes (PCUs) was examined under strong acidic (HNO3) or alkaline (NaClO) oxidative conditions and in presence of a constant strain state. Polyurethane (PU) samples were represented by sheets solvent-cast from commercial pellets or by tubular specimens cut from commercial catheters. The specimens were strained at 100% uniaxial elongation over appropriate extension devices and completely immersed into the oxidative solutions at 50 degrees C for 7-14 days. The changes induced by the oxidative treatments were then evaluated by molecular weight analysis, tensile mechanical tests, and scanning electron microscopy. In the experiments with solvent-cast samples, the PEU Pellethane was degraded more in the alkaline oxidative conditions and mainly in the absence of an applied uniaxial stress. All the tested PCUs were, on the contrary, more affected by the acidic oxidative agent. All the PCUs proved to have overall better stability than the PEU. The susceptibility to oxidation was also dependent on the shape and bulk/surface organisation acquired by the same polymer during its processing. When the oxidative test was applied to catheters made of a PEU and a PCU, the results confirmed the better stability of poly-carbonate-urethanes. PMID- 10794507 TI - In vivo and in vitro stability of modified poly(urethaneurea) blood sacs. AB - In the present study, we investigate the in vivo and in vitro stability of modified poly(urethaneurea) (BioSpan MS/0.4) blood sacs. Blood sacs were utilized primarily in left ventricular assist devices that were implanted in calves for times ranging from 5 to 160 days. Cyclic testing in vitro was also conducted on similar sacs. Various analytical methods were employed to characterize the sacs after in vivo or in vitro service and corresponding retained "control" sacs. These methods included ATR-FTIR spectroscopy, scanning electron microscopy and gel permeation chromatography. In general, the characteristics of implanted and in vitro cycled sacs were similar to their control sacs. Thermal and microtensile properties were unchanged after testing. The same was true for the ATR-FTIR spectra, indicating relative chemical stability for the time frames explored here. The only significant changes occurred in molecular weight and gross surface morphology. A modest increase in weight average molecular weight was observed for most implanted blood sacs, indicating some type of chain extension or branching reaction in vivo. Although the surface morphologies of implanted blood sacs were often similar to their control sacs, we sometimes observed limited pitting on the nonblood contacting surfaces in regions of the sac that experience maximum bending during service. PMID- 10794508 TI - Hydrophilic polymers derived from vitamin E. AB - Vitamin E containing copolymers for biomedical applications were obtained by copolymerization reaction of vitamin E methacrylate (VEMA) with 2-hydroxyethyl methacrylate (HEMA), N,N-dimethyl acrylamide (DMA) or vinyl pyrrolidone (VP), in different experiments. High molecular weight copolymers prepared by free radical reactions initiated by azobisisobutironitrilo, AIBN, present a random distribution of vitamin E derivatives along the macromolecular chains, and the average composition depends on the initial composition of the reaction medium. The relative flexibility of the polymeric systems was analyzed measuring the glass transition temperature of copolymeric sequences and that of the pure alternating diad (Tg12) obtained by the application of the treatments proposed by Johnston and Barton to all the systems. Tg12 was higher than the average Tg of both homopolymers (Tg) for the VEMA-HEMA system, Tg12 was lower than Tg for the VEMA-DMA system and Tg12 was similar to Tg for the VEMA-VP system. VEMA-HEMA copolymers gave rise to hydrogels in water, acidic and alkaline media. VEMA-DMA copolymers gave rise to hydrogels in acidic medium and dissolved in water and alkaline medium. VEMA-VP copolymers were soluble in all media. The swelling of all the hydrogels fit a second-order kinetics. A VEMA-HEMA hydrogel was selected for in vivo experiments in order to study the influence of vitamin E on the regeneration process of Achilles tendon. The polymeric derivatives of vitamin E stimulated the regenerative process as a consequence of the antiaging effect in the local area of application. PMID- 10794509 TI - Liposphere based lipoprotein-mimetic delivery system for 6-mercaptopurine. AB - Long-circulating lipospheres containing 6-mercaptopurine (6-MP) were prepared by solidification of warm microemulsion at low temperature. Palmitoyl PEG was incorporated in the system to confer stealth-type nature. The size of lipospheres was in the range of 60-70 nm and was inversely proportional to sonication time. The size range was attained after 8 h. of sonication. The entrapped 6-MP contained 0.12 mmol/mole of lipid. The coating efficiency of 63-71% was attained. The zeta potential substantially decreased after PEG coating, however, the lipospheres were stable due to steric repulsion and exhibited no aggregation. The release of 6-MP was found to be 18-25% of administered dose in 24 h. and followed a mixed profile for stealth lipospheres. The percent dose remaining in plasma was found to be high even after 24 h as compared to control, indicating an increase in circulation time of lipospheres. Tissue accumulation of drug correlated with the pharmacokinetic behavior of lipospheres. The system seems to be an ideal carrier for anticancer drug delivery. PMID- 10794510 TI - Full-crown castings made with water-saturated and dry casting ring liners. AB - The carcinogenic potential of asbestos has led to the development of non-asbestos materials for use as casting ring liners. Cellulose paper are replacing asbestos paper. The brass die and crown were made to produce full crowns. The standardized wax pattern was replicated twenty-nine times. Two grooves were marked on the die and wax patterns. The marginal gaps between wax pattern and die shoulder were measured. Then, the wax specimens were divided into three groups. The dry ring liner and the water-saturated ring liner were used, in the third group, any liner was used. The castings were fabricated and were fitted on the die. Measurements were made of the distance separating the gingival margin of the castings from the shoulder on the die. Measurement points (X and Y) were the same points used for wax pattern measurement. The difference between wax pattern and casting was found to be statistically significant in all the groups (p = 0.0051). There was no statistically significant difference between groups on the comparative fit of the castings. The mean marginal space in the water-saturated liner group was 0.51 +/- 0.12 mm (X), 0.51 +/- 0.11 mm (Y), and in the dry liner group, the mean marginal space was 0.47 +/- 0.09 mm (X), 0.49 +/- 0.08 mm (Y). In the unlined group, they were 0.52 +/- 0.18 mm (X), 0.55 +/- 0.17 mm (Y). Use of dry ring lining material resulted in smaller marginal spaces between castings and the die than use of water-saturated cellulose ring lining material. The results demonstrated that the use of dry liners may improve the fit of castings to dies. PMID- 10794511 TI - The amount of free corticosterone is increased during lipopolysaccharide-induced fever. AB - The relation between lipopolysaccharide (LPS)-induced fever and bioavailability of corticosterone (B) was examined in male Wistar rats. Animals were injected with LPS (2.5 mg/kg i.p.) or saline and core temperature and heart rate were monitored continuously using a biotelemetry system. Blood samples were withdrawn from freely moving rats via jugular catheters for estimation of total and free plasma B. LPS induced a long-lasting increase (24-48 h) in core temperature and B secretion and a short-lasting increase (90 min) in heart rate. LPS-induced fever was accompanied by a significant increase in the free/total B ratio. In contrast, an acute injection of B, which resulted in circulating B levels similar to those found after LPS, did not affect the free/total B ratio. The important role of LPS induced fever in the hormone secretion pattern and the equilibrium between free and total B was further demonstrated in an in vitro study showing that an increase in the temperature by 3 degrees C elevated the free B fraction and the free/total B ratio of plasma samples with concentrations of B in the physiological range (5-40 microg/dl). Taken together, these findings indicate that during LPS-induced fever there is an increase in the amount of biologically available B. Exposure of glucocorticoid-sensitive targets to elevated levels of free B could contribute to the restoration of homeostasis that is disturbed during inflammation. PMID- 10794512 TI - Hair analysis for drugs of abuse XXI. Effect of para-substituents on benzene ring of methamphetamine on drug incorporation into rat hair. AB - In order to study the effect of para-substituents on the benzene ring of methamphetamine on drug incorporation into hair from blood, the plasma AUCs and hair concentrations of 7 methamphetamines [methamphetamine(MA), p hydroxymethamphetamine(OHMA), p-bromomethamphetamine (BMA), p aminomethamphetamine (AMA), p-nitromethamphetamine (NMA), p methoxymethamphetamine (MOMA) and 3,4-methylenedioxymethamphetamine (MDMA)] plus propylhexedrine(PHX) in DA rats was determined after intraperitoneal injection at 5 mg/kg, with single dose for the plasma AUC and 10 doses for the hair concentration. Drug incorporation rates into hair (ICRs) were calculated by dividing each hair concentration by each plasma AUC. Comparing the highest value (NMA) to the lowest one (OHMA), the ICR of NMA was 31.7 times larger than that of OHMA. Using the ICR of MA which has no substitute on the benzene ring as a base point, nitro, bromo, methylenedioxy, methoxy and amino groups raised the drug incorporation into rat hair in this order. On the other hand, hydroxy substitution showed a negative effect on the ICR. In comparison between the ICRs of MA and PHX, it was found that the benzene ring shows higher affinity to melanin and less lipophilicity than the cyclohexyl ring. Our results showed that there is a relatively strong effect of the functional groups on drug incorporation into hair. The combination of melanin affinity and lipophilicity are clearly correlated with their ICR. PMID- 10794513 TI - Alcohol up-regulates UDP-glucuronosyltransferase mRNA expression in rat liver and in primary rat hepatocyte culture. AB - The interactions between alcohol and cytochrome P-450 enzymes have been well investigated. However, the data regarding the effect of alcohol on the regulation of UDP-glucuronosyltranferase (UGT) activity are less clear. The aim of the present study was to determine the role of alcohol in the regulation of UGT mRNA expression by using whole animal and primary cultured hepatocytes. Chronic ethanol feeding of rats significantly increased the expression of liver UGT1A1 mRNA to 177% of control. The mRNA levels for UGT1A5, UGT2B1 and UGT2B3 were also enhanced, but did not reach statistical significance. In cultured hepatocytes, treatment with either ethanol or isopentanol significantly increased the expression of UGT1A1, UGT1A5, UGT2B1, and UGT2B3 mRNAs, but to different degrees. The induction of UGT1A1 and UGT2B1 mRNAs by ethanol or isopentanol was time dependent and maximal changes occurred at 48 h. The expression of UGT1A6 mRNA was not significantly modified by either ethanol or isopentanol. In conclusion, ethanol and isopentanol have direct roles in the regulation of UGT. PMID- 10794514 TI - Factors regulating amylase secretion from chicken pancreatic acini in vitro. AB - In mammals, cholecystokinin regulates pancreatic exocrine secretion under physiological conditions. We have shown, however, that cholecystokinin at physiological concentrations does not induce pancreatic amylase secretion in birds. Therefore, we investigated the effects of various neurotransmitters and gut hormones on the pancreatic amylase secretory response in isolated chicken pancreatic acini. Acetylcholine (half-maximal stimulation at 800 nM) and vasoactive intestinal polypeptide (half-maximal stimulation at 40 pM) produced a concentration-dependent increase in amylase secretion at physiological concentrations. The combination of acetylcholine and vasoactive intestinal polypeptide produced an additive response in amylase secretion. Sodium nitroprusside, a spontaneous nitric oxide releaser, and bombesin, induced amylase secretion at concentrations greater than 10 nM and 100 nM, respectively. Gastrin and secretin increased amylase secretion at pharmacological concentrations (10 to 100 nM). Our findings suggest that neural regulation is important for pancreatic enzyme secretion in birds and the contribution of gut hormones seems to be physiologically unimportant. PMID- 10794515 TI - Molecular cloning and expression studies of a prolactin receptor in goldfish (Carassius auratus). AB - A full-length cDNA clone, of a size of 4.6 kb, for the goldfish prolactin receptor has been isolated. This cDNA clone encodes a protein of 600 amino acids homologous to prolactin receptors of other species. A Kyte-Doolittle hydropathy analysis of the receptor indicates that the translated protein consists of a signal peptide of 22 amino acids, an extracellular domain of 228 amino acids, a single transmembrane domain of 24 amino acids, and an intracellular domain of 346 amino acids. Several characteristic landmarks of prolactin receptor could be identified in this clone. These include the four conserved cysteine residues and the WS motif within the extracellular domain, and the box 1 and box 2 regions of the intracellular domain. Among all the prolactin receptor sequences known to date, this clone bears the closest resemblance to the tilapia prolactin receptor, although homology between these two fish prolactin receptors is rather low. There are only 57.4% of nucleotide and 48.3% of amino acid sequence identities between these two fish receptors. This receptor cDNA was transfected into CHO-K1 cells for functional analysis. RT-PCR analysis with a pair of gene specific primers indicate that the receptor was transcribed in the transfected cells. Using a cell proliferation assay based on the reduction of the tetrazolium salt 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, the receptor transfected CHO-K1 cells can be stimulated to proliferate upon the addition of ovine prolactin in the culture medium. The tissue distribution of the prolactin receptor in goldfish was studied by RT-PCR/Southern analysis and by Northern analysis. The results indicated that the receptor is expressed mostly in the kidney, the gill and the intestine of goldfish, corroborating with the osmoregulatory role of prolactin in fish. In addition, an appreciable level of the receptor is also found in the brain and gonads of goldfish. Northern analysis showed that there are two transcript sizes, a major 4.6 kb and a minor 3.5 kb mRNAs, in the kidney, gill and intestine. PMID- 10794516 TI - Positive and negative inotropic effects of muscarinic receptor stimulation in mouse left atria. AB - In isolated mouse left atria, acetylcholine (ACh) produced a biphasic inotropic response; a transient decrease in developed tension was followed by an increase. Both negative and positive responses were concentration dependent and were inhibited by atropine. The negative and positive inotropic responses were also observed with a nonselective muscarinic stimulant, oxotremorine-M, but not with an M1-receptor selective stimulant, McN-A343. Pirenzepine, an M1-receptor antagonist, inhibited both negative and positive inotropic responses at high concentrations. Gallamine, an M2-receptor antagonist, inhibited the negative response. Hexahydro-siladifenidol hydrochloride, p-fluoro analog (p-F-HHSiD), an M3-receptor antagonist, inhibited the positive response with no effect on the negative phase. In pertussis toxin (PTX) treated preparations, negative inotropic response to ACh was not observed. These results suggest that the negative and positive inotropic responses to acetylcholine in mouse atria are mediated by M2 and M3 receptors, respectively. The negative phase, but not the positive phase, was mediated by a PTX-sensitive G protein. PMID- 10794517 TI - Diabetes influences the effect of 17beta-estradiol on mechanical responses of rat urethra and detrusor strips. AB - Estrogen deficiency is one of the factors involved in the stress incontinence in postmenopausal women, and estrogens have been used clinically in the treatment of urinary disorders during menopause. Sex hormones seem to be also involved in the diabetic changes of urinary bladder and urethra, because ovariectomy causes an increase in the micturition of streptozotocin-diabetic rats. In the present study diabetic and healthy female rats were used to investigate the effect of 17beta estradiol on mechanical contractions to norepinephrine and to KCI and relaxations to ATP on isolated proximal urethral preparations as well as on contractions to ACh, ATP and KCl on detrusor smooth muscle strips. The data were compared with those obtained in OVX animals, with or without estradiol replacement. The present study showed that ovariectomy decreased the responses to ATP, NE and KCl in urethral preparations, and responses to ATP, ACh and KCl in bladder strips from both healthy and diabetic rats. Diabetes appeared to potentiate the effect of ovariectomy in both tissues. Estrogen replacement was able to recover functional responses in urethras of healthy rats. In diabetic rats, this treatment partially restored ATP-induced responses in both tissues, almost completely restored those to NE in urethra and those to ACh in bladder. This study clearly indicated that abnormalities of urethra and bladder function caused by ovariectomy can be restored by estrogen treatment also in diabetic animals, at least at an early stage of disease. PMID- 10794518 TI - Four-week ethanol drinking increases both thyrotropin-releasing hormone (TRH) release and content in rat pancreatic islets. AB - Ethanol exerts profound effects on the endocrine and exocrine pancreas. Some effects of chronic alcohol consumption on insulin secretion in response to glucose load are similar to those of TRH gene disruption. TRH is present in insulin-producing B-cells of the islets of Langerhans; its role in this location is still not fully explored. To examine the possible effect of long-term in vivo ethanol treatment on pancreatic TRH we compared three groups of rats: a 10% (wt:vol) ethanol-drinking group (E), absolute controls (AC) and pair-fed (PF) group with solid food intake corresponding to that of E. The fluidity of pancreatic membranes was not affected by chronic in vivo exposure of rats to ethanol, but was significantly decreased in PF group. Four-week treatment resulted in significantly higher TRH content in isolated islets of the E group and increased basal and 80 mM isotonic ethanol-induced secretion compared to AC and PF. Plasma levels of insulin, C-peptide, IGF-I, and glycemia were, however, not affected by ethanol treatment. Cell swelling, which can be induced by the presence of permeants (e.g. ethanol) in an isotonic extracellular medium, is a strong stimulus for secretion in various types of cells. In the present study, isosmotic ethanol (40, 80, and 160 mM) induced dose-dependent release of TRH and insulin from adult rat pancreatic islets in vitro. The same concentrations were not effective when applied in a hyperosmotic medium (addition of ethanol directly to the medium), thus indicating the participation of cell swelling in the ethanol induced secretion. In conclusion, chronic ethanol treatment significantly affected pancreatic TRH and this effect might be mediated by cell swelling. The role of these changes in the profound effect of ethanol on the endocrine and exocrine pancreas remains to be established. PMID- 10794519 TI - Expression of glucose transporter 4 mRNA in adipose tissue and skeletal muscle of ovariectomized rats treated with sex steroid hormones. AB - The effects of 17beta-estradiol (E) and/or progesterone (P) on glucose transporter 4 (GLUT4) expression in the adipose tissue and skeletal muscle of ovariectomized female rats were studied. The Sprague-Dawley rats received daily subcutaneous injections of various doses of E and/or P for 7 days (n=5-6 per dose). The expression of GLUT4 mRNA was assessed by performing ribonuclease protection assays. GLUT4 protein levels were assessed by Western blotting assays. The adipose tissue levels of GLUT4 mRNA were reduced by the administration of 50 microg E, which resulted in unphysiologically high serum E concentrations. Although the GLUT4 mRNA levels did not change after the administration of 10 microg E or 5 mg P, they were reduced significantly to approximately half the control group level by the administration of both hormones (p <0.01). The skeletal muscle GLUT4 mRNA levels were not changed significantly by hormone treatment. These findings suggest that E and P may be involved in the regulation of GLUT4 mRNA expression in adipose tissue. PMID- 10794520 TI - Brain distribution of 6-mercaptopurine is regulated by the efflux transport system in the blood-brain barrier. AB - 6-mercaptopurine (6-MP) has been used clinically for 40 years to maintain remission in patients with acute lymphoblastic leukemia (ALL). However, central nervous system (CNS) relapses frequently occur in patients with ALL who continuously receive anticancer drugs, including 6-MP, during remission maintenance therapy. The cause of such CNS relapse is not well understood. One possible reason may involve the restricted distribution of 6-MP in the brain. This study, therefore, investigates the blood-brain barrier (BBB) transport which largely regulates 6-MP distribution in the brain using a quantitative microdialysis technique and centers on the efflux transport of 6-MP across the BBB. The brain tissue, cerebrospinal fluid (CSF), or hippocampal interstitial fluid (ISF) concentration of 6-MP was very low compared with the unbound plasma concentration, suggesting that 6-MP distribution in the brain is highly restricted. Kinetic analyses of this BBB transport showed that the efflux clearance from brain ISF to plasma across the BBB (CLout) is approximately 20 times greater than the influx clearance from plasma to brain (CLin). The CLout was significantly reduced by 1mM N-ethylmaleimide (NEM), a sulfhydryl-modifying agent, suggesting the participation of transport protein in the efflux of 6-MP across the BBB. In addition, efflux transport was inhibited by an intracerebral infusion of probenecid (1.5 mM), p-aminohippuric acid (PAH, 3.0 mM), benzoate (3.6 mM), or salicylate (3.7 mM) administered through a microdialysis probe, but neither choline (0.8 mM) nor tetraethylammonium (TEA, 0.7 mM) had any effect. These data suggest that the restricted 6-MP brain distribution may be ascribed to efficient efflux from the brain, possibly via both the organic anion transport system, shared with probenecid and PAH, and the monocarboxylic acid transport system, shared with benzoate and salicylate. PMID- 10794521 TI - Pharmacological modulation of fluid secretion in the pigmented rabbit conjunctiva. AB - We determined net fluid secretion rate across the pigmented rabbit conjunctiva in the presence and absence of pharmacological agents known to affect active Cl- secretion and Na+ absorption. Fluid flow across a freshly excised pigmented rabbit conjunctiva mounted between two Lucite half chambers was measured by a pair of capacitance probes in an enclosed cabinet maintained at 37 degrees C and a relative humidity of 70%. Fluid transport was also measured in the presence of compounds known to affect active Cl- secretion (cAMP, UTP, and ouabain), Na+ absorption (D-glucose), or under the Cl--free condition on both sides of the tissue. Net fluid secretion rate across the pigmented rabbit conjunctiva in the serosal-to-mucosal direction at baseline was 4.3+/-0.2 microl/hr/cm2 (mean +/- s.e.m.). Net fluid secretion rate was increased approximately two-fold by mucosally applied 1 mM 8-Br cAMP (8.4+/-0.4 microl/hr/cm2) and 10 microM UTP (9.8+/-0.6 microl/hr/cm2), but was abolished by either serosally applied 0.5 mM ouabain (0.3+/-0.1 microl/hr/cm2) or under the Cl--free conditions (0.06+/-0.04 microl/hr/cm2). Mucosal addition of 20 mM D-glucose decreased net fluid secretion rate to 1.0+/-0.5 microl/hr/cm2. In conclusion, the pigmented rabbit conjunctiva appears to secrete fluid secondary to active Cl- secretion. This net fluid secretion is subject to modulation by changes in active Cl- secretion rate and in mucosal fluid composition such as glucose concentration. PMID- 10794522 TI - Differential upregulation of p53-responsive genes by genotoxic stress in hematopoietic cells containing wild-type and mutant p53. AB - Cells respond to genotoxic stress by activation of many genes, including the tumor suppressor p53. p53 activates transcriptionally target genes, such as p21waf1 and gadd45, which can lead to cell cycle arrest, or bax, which can lead to cell death. We examined the response to genotoxic stress in two hematopoietic cell lines that harbor either wild-type (MOLT-4) or a mutant p53 with a codon 161 mutation (U266). We adapted a multiprobe RNase protection assay (RPA) to determine the steady-state RNA levels, and in combination with nuclear runoff assays, transcriptional rates of multiple stress-induced genes. We found a differential activation of growth arrest and cell death-specific p53 target genes in cells with wild-type or mutant p53. Our results show that genotoxic stress can activate the p21waf1 and gadd45 genes in both cell lines. However, the bax gene was not induced in U266 cells. Bax and gadd45 gene induction could be efficiently blocked by pretreating the cells with the antioxidant compound pyrrolidine dithiocarbamate, suggesting that oxidative stress was involved in these responses. Induction of all three genes in MOLT-4 cells was clearly at the transcriptional level, because we detected transcriptional activity by nuclear runoff RPA assays, and transfection with a consensus p53 binding sequence. U266 cells did not activate the same reporter, in spite of the upregulation of p21waf1 and gadd45 RNA levels. However, the p21waf1-reporter constructs containing 0.9 to 2.4 kb of the native p21 promoter were potently activated in U266 cells. These results indicate a differential regulation of p53 target genes in cells containing wild-type or codon 161 mutant p53. PMID- 10794523 TI - LIM domain-containing protein trip6 can act as a coactivator for the v-Rel transcription factor. AB - The retroviral oncoprotein v-Rel is a transcriptional activator in the Rel/NF kappaB family of eukaryotic transcription factors. v-Rel malignantly transforms a variety of cell types in vitro and in vivo, and its transforming activity is dependent on the ability of v-Rel to bind to DNA and activate transcription. In this report, we used the yeast two-hybrid assay to identify proteins that interact with C-terminal sequences of v-Rel that are needed for transcriptional activation and transformation. One protein, Trip6, that we identified in this screen was previously identified as a thyroid hormone receptor-interacting protein. Trip6 is a member of a subfamily of LIM domain-containing proteins that are thought to transport intracellular signals from the cell surface to the nucleus. By several criteria, we show that sequences from Trip6, which include the LIM domains, behave as a coactivator for transcriptional activation by v-Rel. That is, a GAL4-Trip6 fusion protein can activate transcription in yeast and chicken cells, Trip6 can enable C-terminal sequences of v-Rel to activate transcription in yeast, and Trip6 can enhance activation by v-Rel from a kappaB site reporter plasmid in yeast. Although full-length Trip6 localizes to adhesion plaques, deletion of N-terminal sequences allows human Trip6 to enter the nucleus of chicken cells. Lastly, Northern blotting shows that Trip6 mRNA is expressed in many human tissues. Coexpression of Trip6 does not affect the transforming activity of v-Rel. Taken together, our results indicate that Trip6 may be a protein that is important for the ability of v-Rel to activate transcription and transform cells, and may represent a potential target for blocking Rel-mediated oncogenesis and transcriptional activation. PMID- 10794524 TI - Rel/NF-kappaB represses bcl-2 transcription in pro-B lymphocytes. AB - The mechanisms controlling programmed cell death (PCD) during early B cell development are not well understood. Members of both the Bcl-2 family of apoptosis-related proteins and the nuclear factor-kappa B/Rel (NF-kappaB/Rel) family of transcription factors are expressed differentially during B cell development. To date, however, no direct interactions between these two families have been demonstrated. The FL5.12 cell line represents a model for progenitor B cell development. Such cells reproducibly undergo PCD upon IL-3 withdrawal. The signal to enter the apoptotic pathway is mediated by a shift in the ratio of Bcl 2:Bax. While bax levels remain constant, bcl-2 transcription rate, steady-state mRNA, and protein levels decrease. Analysis of the bcl-2 promoter reveals 3 kappaB sites functionally able to bind kappaB factors from FL5.12 nuclear extracts. Cotransfection studies demonstrate that NF-kappaB factors can repress bcl-2 transcription and that site-directed mutagenesis of the kappaB motifs abolishes this repression. These studies suggest that NF-kappaB mediates PCD in pro-B cells through transcriptional repression of the survival gene bcl-2, thus shifting the bcl-2:bax ratio in favor of death-promoting complexes. PMID- 10794525 TI - Cloning and expression of the mouse deoxyuridine triphosphate nucleotidohydrolase gene: differs from the rat enzyme in that it lacks nuclear receptor interacting LXXLL motif. AB - We have previously reported the cloning of rat deoxyuridine triphosphate nucleotidohydrolase (dUTPase) cDNA and demonstrated that the full-length protein as well as the N-terminal 62-amino acid peptide interacts with peroxisome proliferator-activated receptor alpha (PPARalpha). We now report the cloning of mouse dUTPase cDNA and show that it contains a 162-amino acid open reading frame, encoding a protein with a predicted Mr of 17,400 and differs from rat cDNA, which contains additional 43 amino acids at the N-terminal end. Unlike rat dUTPase, mouse dUTPase failed to bind PPARalpha. An evaluation of 205 amino acid containing rat dUTPase cDNA revealed that the N-terminal 43 extra amino acid segment contains an LXXLL signature motif, considered necessary and sufficient for the binding of several cofactors with nuclear receptors, and its absence in murine dUTPase possibly accounts for the differential binding of these enzymes to PPARalpha. In situ hybridization and immunohistochemical studies revealed that, in the adult mouse, dUTPase is expressed at high levels in proliferating cells of colonic mucosa, and of germinal epithelium in testis. At 9.5-day mouse embryonic development, dUTPase expression is predominantly in developing neural epithelium, and hepatic primordium, and in later developmental stages (11.5-, 13.5-, and 15.5 day embryo), the expression began to be localized to the liver, kidney, gut epithelium, thymus, granular layer of the cerebellum, and olfactory epithelium. We also show that the murine dUTPase gene comprises 6 exons and the 5'-flanking region of -1479 to -27, which exhibited high promoter activity, contains a typical TATA box and multiple cis-elements such as Sp-1, AP2, AP3, AP4, Ker1, RREB, and CREB binding sites. These observations suggest the existence of variants of dUTPase, some of which may influence nuclear receptor function during development and differentiation, in addition to catalyzing the hydrolysis of dUTP to dUMP. PMID- 10794527 TI - His-Asp phosphorelay signaling: a communication avenue between plants and their environment. AB - His-Asp phosphorelay systems have been recently discovered in plants and have emerged as some of the most important signaling systems. The phosphorelay systems in plants include components with sensor (His-protein kinase) domains, His containing phosphotransfer (HPt) domains, and receiver (response regulator) domains. Recent studies implicate phosphorelay systems in sensing and propagating signals from a wide variety of external and/or internal stimuli such as ethylene, cytokinin, and osmolarity. In maize and Arabidopsis, some response regulators are up-regulated by both cytokinins and nitrate. These findings imply that the His Asp phosphorelay may operate in an inorganic nitrogen-signaling pathway mediated by cytokinin in plants. PMID- 10794526 TI - The myogenic regulatory circuit that controls cardiac/slow twitch troponin C gene transcription in skeletal muscle involves E-box, MEF-2, and MEF-3 motifs. AB - We have characterized the specific DNA regulatory elements responsible for the function of the human cardiac troponin C gene (cTnC) muscle-specific enhancer in myogenic cells. We used functional transient transfection assays with deletional and site-specific mutagenesis to evaluate the role of the conserved sequence elements. Gel electrophoresis mobility shift assays (EMSA) demonstrated the ability of the functional sites to interact with nuclear proteins. We demonstrate that three distinct transcription activator binding sites commonly found in muscle-specific enhancers (a MEF-2 site, a MEF-3 site, and at least four redundant E-box sites) all contribute to full enhancer activity but a CArG box does not. Mutation of either the MEF-2 or MEF-3 sites or deletion of the E-boxes reduces expression by 70% or more. Furthermore, the MEF-2 site and the E-boxes specifically bind, respectively, to MEF-2 and myogenic determination factors derived from nuclear extracts. EMSA assays using a MEF-3 containing oligonucleotide revealed indistinguishable separation patterns with extracts from myogenic cells and nonmyogenic cells. These data suggest that expression of the cTnC gene in slow-twitch skeletal muscle is sustained through complex interactions at the 3'Ile enhancer between muscle-specific and nontissue-specific transcription factors: either a myogenic bHLH complex or MEF-2 can activate transcription but only in the presence of a third transcriptional activator that appears not to be muscle specific. We conclude from these observations that the cTnC 3'Ile element is a composite enhancer that functions through the combined interactions of at least five regulatory elements and their cognate binding factors: three or four E-boxes, a MEF-2 site, and a MEF-3 site. The data support the notion that all of these sites contribute to enhancer function in cell systems in an additive way but that none are absolutely required for enhancer activity. The data imply that the levels of transcription of cTnC in myogenic tissues in which the activities of one of the transcriptional factors is lacking would be partially but not wholly suppressed. Our data support the critical role of E-box sites in conjunction with the adjacent elements. Hence, we assign CTnC gene regulation to the "ordinary" rather than to the "novel" category of transcriptional regulation during skeletal myogenesis. PMID- 10794528 TI - AtZFP1, encoding Arabidopsis thaliana C2H2 zinc-finger protein 1, is expressed downstream of photomorphogenic activation. AB - C2H2 zinc-finger proteins play important roles in plant development including floral organogenesis, leaf initiation, lateral shoot initiation, gametogenesis and seed development. The gene for one such protein from Arabidopsis, AtZFP1 (Arabidopsis thaliana zinc-finger protein 1), is expressed at high levels in the shoot apex, including the apical meristem, developing leaves and the developing vascular system. In light-grown seedlings, AtZFP1 expression is induced about three days after germination, before the expansion of the true leaves. Dark-grown plants, in which photomorphogenesis is repressed, have no detectable AtZFP1 expression in the shoot apex. Under conditions which induce or mimic photomorphogenic development including growth in the light, shifting dark-grown plants to continuous light or growth on cytokinin in the dark, high levels of AtZFP1 expression are detected. Furthermore, AtZFP1 expression does not depend on active photosynthesis as shown by analysis of plants grown on the carotenoid biosynthetic inhibitor norflurazon. These results are discussed in relation to a possible role for AtZFP1 in shoot development, downstream of photomorphogenic activation. PMID- 10794529 TI - A LIM-domain protein from sunflower is localized to the cytoplasm and/or nucleus in a wide variety of tissues and is associated with the phragmoplast in dividing cells. AB - LIM proteins are important eucaryotic developmental regulators characterized by the presence of one or several double zinc finger motifs, the LIM domains, which are protein-interacting domains. Using the cDNA of the previously described pollen LIM protein PLIM1 from sunflower as a hybridization probe we have isolated the coding sequence for a related protein from cDNA libraries from various sunflower organs. This protein, WLIM1, is 188 amino acids long and, like the pollen protein PLIM1, contains two LIM domains, separated by a 48 residue spacer region. The two sunflower proteins are structurally related to the animal LIM proteins CRP and MLP. A WLIM1 gene transcript was detected by RT-PCR in all vegetative and reproductive plant organs tested. Polyclonal antibodies raised against the bacterially expressed and affinity-purified protein recognize a polypeptide of ca. 50 kDa in these organs. Immunocytochemical studies detect the protein in many cell types in each of these organs where it is localized either to the cytoplasm, the nucleus, or both. The protein is often associated with plastids and smaller cellular structures or organelles. In late anaphase and early telophase of dividing cells from ovaries, stems and roots it accumulates in the phragmoplast, and may therefore also play a role in cytokinesis. PMID- 10794530 TI - Characterization of the LI818 polypeptide from the green unicellular alga Chlamydomonas reinhardtii. AB - The LI818 gene from Chlamydomonas encodes a polypeptide that is related to the chlorophyll a/b-binding proteins (CAB) of higher plants and green algae. However, despite this relatedness, LI818 gene expression is not coordinated with that of cab genes and is regulated differently by light, suggesting a different role for LI818 polypeptide. We show here that, in contrast to CAB polypeptides, LI818 polypeptide is not tightly embedded into the thylakoid membranes and is localized in stroma-exposed regions. Moreover, during chloroplast development, LI818 polypeptide accumulates before CAB polypeptides. We also show that the LI818 polypeptide forms with certain chlorophyll a/c-binding proteins (CAC) from the haptophyte Isochrysis galbana and the diatom Cyclotella cryptica a natural group that is distinct from those constituted by CAB, CAC and the chlorophyll a/a binding proteins (CAA). Such an association suggests a very ancient origin for this group of polypeptides, which predates the division of the early photosynthetic eukaryotes into green (chlorophyte), red (rhodophyte) and brown (chromophyte) algae. Possible roles for the LI818 polypeptide are discussed. PMID- 10794531 TI - Polygalacturonase gene expression in kiwifruit: relationship to fruit softening and ethylene production. AB - In kiwifruit, much of the softening process occurs prior to the respiratory climacteric and production of ethylene. This fruit therefore represents an excellent model system for dissecting the process of softening in the absence of endogenous ethylene production. We have characterized the expression of three polygalacturonase (PG) cDNA clones (CkPGA, B and C) isolated from fruit of Actinidia chinensis. Expression of CkPGA and B was detected by northern analysis only in fruit producing endogenous ethylene, and by RT-PCR in other tissues including flower buds, petals at anthesis, and senescent petals. CkPGA promoter fragments of 1296, 860 and 467 bp fused to the beta-glucuronidase (uidA) reporter gene directed fruit-specific gene expression during the climacteric in transgenic tomato. CkPGC gene expression was observed in softening fruit, and reached maximum levels (50-fold higher than for CkPGA and B) as fruit passed through the climacteric. However, expression of this gene was also readily detected during fruit development and in fruit harvested prior to the onset of softening. Using RT-PCR, expression of CkPGC was also detected at low levels in root tips and in senescent petals. These results suggest that PG expression is required not only during periods of cell wall degeneration, but also during periods of cell wall turnover and expansion. PMID- 10794532 TI - Identification of a novel four-domain member of the proteinase inhibitor II family from the stigmas of Nicotiana alata. AB - Proteinase inhibitors (PIs) of the potato type II family have been identified in a number of solanaceous species. Most family members have two PI domains which are specific for either chymotrypsin or trypsin. More recently family members have been described with three or six repeated PI domains. Here we describe a novel four-domain family member produced in the stigmas and leaves of the ornamental tobacco, Nicotiana alata, which has high sequence identity with a six domain member from the same species. Both proteins are produced as precursors that enter the secretory pathway and are subsequently processed into a series of 6 kDa Pis. The four- and six-domain precursor proteins were isolated from immature stigmas and characterised by mass spectrometry which revealed that both proteins had been trimmed at the N-terminus, at a position corresponding to the predicted signal peptide cleavage site. Furthermore, no post-translational modifications were apparent. PMID- 10794533 TI - The yeast polyadenylate-binding protein (PAB1) gene acts as a disease lesion mimic gene when expressed in plants. AB - We have expressed the gene (PAB1) encoding the yeast polyadenylate-binding protein (Pab1p) in tobacco. Plants that accumulate the Pab1p display a range of abnormalities, ranging from a characteristic chlorosis in leaves to a necrosis and large inhibition of growth. The severity of these abnormalities reflects the levels of yeast Pab1p expression in the transgenic plants. In contrast, no obvious differences could be seen in callus cultures between the transgene and vector control. Plants that display PAB-associated abnormalities were resistant to a range of plant pathogens, and had elevated levels of expression of a pathogenesis-related gene. These two properties--impairment of growth and induction of defense responses--indicate that the yeast PAB1 gene can act as a disease lesion mimic gene in plants. PMID- 10794534 TI - An Arabidopsis thaliana protein homologous to cyanobacterial high-light-inducible proteins. AB - An Arabidopsis thaliana cDNA clone encoding a novel 110 amino acid thylakoid protein has been sequenced. The in vitro synthesized protein is taken up by intact chloroplasts, inserted into the thylakoid membrane and the transit peptide is cleaved off during this process. The mature protein is predicted to contain 69 amino acids, to form one membrane-spanning alpha-helix and to have its N-terminus at the stromal side of the thylakoid membrane. The protein showed similarity to the LHC, ELIP and PsbS proteins of higher plants, but more pronounced to the high light-inducible proteins (HLIPs) of cyanobacteria and red algae, to which no homologue previously has been detected in higher plants. As for HLIP and ELIP, high light increases the mRNA levels of the corresponding gene. Sequence comparisons indicate that the protein may bind chlorophyll and form dimers in the thylakoid membrane. The level of expression of the protein seems to be far lower than that of normal PSI and PSII subunits. PMID- 10794535 TI - Characterization of chloroplast psbA transformants of Chlamydomonas reinhardtii with impaired processing of a precursor of a photosystem II reaction center protein, D1. AB - One of the photosystem II reaction center proteins, D1, is encoded by the psbA gene and is synthesized as a precursor form with a carboxyl-terminal extension that is subsequently cleaved between Ala-344 and Ser-345. We have generated three psbA transformants of the green alga Chlamydomonas reinhardtii in which Ala-344 or Ser-345 have been substituted with Pro or Glu (A344P, S345E, and S345P) to understand the effects of the amino acid substitutions on the processing of the precursor D1. S345E grew photoautotrophically and showed PSII activity like the wild type. However, A344P and S345P were unable to grow photoautotrophically and were significantly photosensitive. A344P was deficient in the processing of precursor D1 and in oxygen-evolving activity, but assembled photosystem II complex capable of charge separation. In contrast, both precursor and mature forms of D1 accumulated in S345P cells from the logarithmic phase and the cells evolved oxygen at 18% of wild-type level. However, S345P cells from the stationary phase contained mostly the mature D1 and showed a twofold increase in oxygen-evolving activity. The rate of processing of the accumulated pD1 was estimated to be about 100 times slower than in the wild type. It is therefore concluded that the functional oxygen-evolving complex is assembled when the precursor D1 is processed, albeit at a very low rate. These results suggest the functional significance of the amino acid residues at the processing site of the precursor D1. PMID- 10794536 TI - Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana. AB - Mevalonate kinase (MVK), the enzyme that catalyzes the phosphorylation of mevalonate to produce mevalonate 5-phosphate, is considered as a potential regulatory enzyme of the isoprenoid biosynthetic pathway. The Arabidopsis thaliana MVK gene corresponding to the MVK cDNA previously isolated has been cloned and characterized. RNAse protection analysis indicated that the expression of the MVK gene generates three mRNA populations with 5' ends mapping 203, 254 and 355 nt upstream of the MVK ATG start codon. Northern blot analysis showed that the MVK mRNA accumulates preferentially in roots and influorescences. Histochemical analysis, with transgenic A. thaliana plants containing a translational fusion of a 1.8 kb fragment of the 5' region of the MVK gene to the beta-glucuronidase (GUS) reporter gene, indicated that the MVK 5'-flanking region directs widespread expression of the GUS gene throughout development, although the highest levels of GUS activity are detected in roots (meristematic region) and flowers (sepals, petals, anthers, style and stigmatic papillae). The expression pattern of the MVK gene suggests that the role of the encoded MVK is the production of a general pool of mevalonate-5-phosphate for the synthesis of different classes of isoprenoids involved in both basic and specialized plant cell functions. Functional promoter deletion analysis in transfected A. thaliana protoplasts indicated that regulatory elements between positions -295 and -194 of the MVK 5'-flanking region are crucial for high-level MVK gene expression. PMID- 10794537 TI - Cloning of beta-1,3-glucanases expressed during Cichorium somatic embryogenesis. AB - Three different beta-1,3-glucanase cDNA fragments, CG1, CG2 and CG3, were obtained by RT-PCR from RNA isolated from Cichorium hybrid '474' leaf fragments cultured for 11 days under somatic embryogenesis-inducing conditions. When expressed in Escherichia coli the proteins encoded by the three cDNAs were recognized by antibodies raised against 38 kDa extracellular beta-1,3-glucanases studied previously (Helleboid et al., Planta 205 (1998) 56-63). The CG2 and CG3 cDNAs may represent expressed alleles of one gene because their sequences showed a very high identity (98.5%) and are only 70% identical with CG1. Southern blot analysis revealed the presence of 3-4 genes coding for beta-1,3-glucanases in the Cichorium genome. Expression analysis of the genes corresponding to the three clones analysed by semi-quantitative RT-PCR indicated that CG1 mRNAs were only detectable in Cichorium hybrid '474' leaf fragments from day 3 of somatic embryogenesis induction, whereas CG2-CG3 mRNAs were already present in non induced leaf tissue of both the embryogenic hybrid '474' and a non-embryogenic genotype. The level of CG1 mRNAs was particularly high when embryogenic cells were dividing to produce embryos, and when the amount of callose deposited in cell walls surrounding embryogenic cells and young embryos decreased. These results indicate that expression of the CG1 gene is correlated to the somatic embryogenesis process and that it encodes a 38 kDa beta-1,3-glucanase protein that may be involved in the degradation of callose localized around embryogenic cells and young embryos. A full-length CG1 cDNA clone was obtained using 3' and 5' RACE-PCR, and its sequence revealed that it encodes a beta-1,3-glucanase that is equally homologous to both class III and class IV plant beta-1,3-glucanases. PMID- 10794538 TI - Isolation and characterization of two pathogen- and salicylic acid-induced genes encoding WRKY DNA-binding proteins from tobacco. AB - A pathogen- and salicylic acid (SA)-induced DNA-binding activity has been recently identified in tobacco that is related to a previously identified class of WRKY DNA-binding proteins. To identify members of the WRKY gene family associated with this DNA-binding activity, we have attempted to isolate those WRKY genes that are induced by pathogen infection. Using a domain-specific differential display procedure, we have isolated two tobacco WRKY genes, tWRKY3 and tWRKY4, that are rapidly induced in resistant tobacco plants after infection by tobacco mosaic virus (TMV). Both tWRK3 and tWRKY4 encode proteins with a single WRKY domain that contain the conserved WRKYGQK sequence. Unlike other isolated WRKY proteins that contain the Cys2His2 zinc motif, tWRKY3 and tWRKY4 appear to contain the Cys2HisCys zinc motif. Nonetheless, both tWRKY3 and tWRKY4 are capable of binding DNA molecules with the W-box (TTGAC) element recognized by other WRKY proteins. Expression of the tWRKY3 and tWRKY4 genes could be rapidly induced not only by TMV infection but also by SA or its biologically active analogues that are capable of inducing pathogenesis-related genes and enhanced resistance. Interestingly, induction of both genes by TMV infection was still observed in resistant tobacco plants expressing the bacterial salicylate hydroxylase gene (nahG), although the levels of induction appeared to be reduced. Identification of pathogen- and SA-induced genes encoding WRKY DNA-binding proteins should facilitate future studies on the regulation and functions of this novel group of DNA-binding proteins. PMID- 10794540 TI - The preeclampsia enigma and the renin-angiotensin system. PMID- 10794541 TI - Structural symmetry of the metabolic reaction network. I. Carboxylic acid metabolism. AB - A new approach to systematization of the information on metabolism, based on the symmetry observed in the pattern of the metabolic reaction network, is discussed. A theoretical substantiation of symmetry existence is given. A symmetrical metabolic scheme is developed for metabolism of carboxylic acids. The symmetry of the metabolic reaction network can be used for systematization of biochemical, physiological, medical and other data associated with metabolism as well as for prediction of the new information. PMID- 10794539 TI - Regulation of programmed cell death in maize endosperm by abscisic acid. AB - Cereal endosperm undergoes programmed cell death (PCD) during its development, a process that is controlled, in part, by ethylene. Whether other hormones influence endosperm PCD has not been investigated. Abscisic acid (ABA) plays an essential role during late seed development that enables an embryo to survive desiccation. To examine whether ABA is also involved in regulating the onset of PCD during endosperm development, we have used genetic and biochemical means to disrupt ABA biosynthesis or perception during maize kernel development. The onset and progression of cell death, as determined by viability staining and the appearance of internucleosomal DNA fragmentation, was accelerated in developing endosperm of ABA-insensitive vp1 and ABA-deficient vp9 mutants. Ethylene was synthesized in vp1 and vp9 mutant kernels at levels that were 2-4-fold higher than in wild-type kernels. Moreover, the increase and timing of ethylene production correlated with the premature onset and accelerated progression of internucleosomal fragmentation in these mutants. Treatment of developing wild type endosperm with fluridone, an inhibitor of ABA biosynthesis, recapitulated the increase in ethylene production and accelerated execution of the PCD program that was observed in the ABA mutant kernels. These data suggest that a balance between ABA and ethylene establishes the appropriate onset and progression of programmed cell death during maize endosperm development. PMID- 10794542 TI - Clinical implications of dysregulated cytokine production. AB - Cytokines are soluble proteins that are produced and secreted as part of the immune response to a variety of tissue insults including infection, cancer, and autoimmunity. Most cytokines are secreted by cells of the immune system, but some (for example, type I interferons) are released from "nonimmunological" cells such as fibroblasts and epithelial cells. Cytokines have pleiotropic effects, acting on many somatic cell types to modulate the host's immune response. For the most part, cytokines exert their antimicrobial actions locally---they are secreted by cells in the area of infection, and their effects are restricted to neighboring cells. While many of their local effects benefit the host, cytokines are soluble molecules that may act systemically and are often responsible for many of the symptoms of infection (e.g., headache, fever, myalgia). In high concentrations they can be toxic, or even lethal. Human clinical trials involving the systemic injection of purified cytokines such as interleukins 2 and 12 and tumor necrosis factor alpha provide compelling evidence for the toxicity of these molecules. Likewise, studies of septic shock syndrome demonstrate how overproduction/aberrant production of inflammatory cytokines can lead to rapid mortality. The host may attempt to counter high cytokine levels by releasing soluble cytokine receptors (sCR) or by synthesizing high-affinity anti-cytokine antibodies (acAb), and these natural responses have spawned great interest as potential therapeutic approaches for alleviating cytokine-mediated disease. However, recent studies indicate that these in vivo interactions are much more complex than previously realized; administration of sCR or acAb may either inhibit or (paradoxically) enhance cytokine activity. An alternative therapeutic approach is to intervene at the source of cytokine production. T cells initiate cytokine production only upon antigen contact and terminate synthesis almost immediately after this contact is broken. Thus T cells secrete cytokines specifically at sites of infection and do not continuously produce these potentially toxic molecules while migrating through uninfected tissues or the bloodstream. By learning more about the molecular mechanisms involved with on/off regulation of cytokine production we may be able to develop novel therapeutic drugs to protect against cytokine-mediated immunopathology. This review discusses the regulation of cytokine function by sCR and acAb and compares this to the regulatory mechanisms that are associated with antigen-specific cytokine release by T cells. PMID- 10794543 TI - Identification of two polymorphisms in the early growth response protein-1 gene: possible association with lipid variables. AB - Early growth response factor (EGR)-1 may play an important role in the development of atherosclerosis by inducing the expression of several relevant genes which contribute to the complex modulation of vascular structure and function, leading to vascular occlusive lesions. To investigate the possible role of molecular variants in the human EGR-1 gene for the predisposition to atherosclerosis or coronary heart disease we screened the 5'- and 3'- flanking regions and the entire coding sequence for polymorphisms by polymerase chain reaction/single-strand conformation polymorphism analysis and sequencing. Male patients (n=615) with myocardial infarction and 720 age-matched, male control subjects of the Etude Cas-Temoin de l'Infarctus du Myocarde were genotyped for two newly identified polymorphisms in the 5'- (C-151T) and 3'- (T+861C) flanking region of the EGR-1 gene using hybridization with allele-specific oligonucleotides. Allele and genotype frequencies did not significantly differ between patients with myocardial infarction and control subjects without coronary heart disease. In controls not taking hypolipidemic drugs there was a significant association of the -151T allele with lower plasma levels of total cholesterol (P=0.029), low-density lipoprotein cholesterol (P=0.025) and apolipoprotein B (P=0.038) and a higher ratio of high-density to low-density lipoprotein (P=0.049) than with the C-151 allele. We conclude that the C-151T polymorphism of the EGR-1 gene may contribute to modifications of the lipid metabolism. Our findings need to be replicated in independent studies, and in vitro promoter studies should evaluate the functional consequence of the -151T allele, which disrupts a consensus core sequence for the ubiquitous transcription factor activator protein 4. PMID- 10794544 TI - Beta1-adrenoceptor gene variations: a role in idiopathic dilated cardiomyopathy? AB - A substantial body of evidence suggests involvement of the human beta1 adrenoceptor (beta1-AR) gene in the pathophysiology of dilated cardiomyopathy (DCM), a severe heart disease of significant public health impact. Beta1-AR mediated signal transduction is dramatically altered due to downregulation, resulting in an impairment of myocardial response. The important role of genetic factors in idiopathic dilated cardiomyopathy (IDCM) recently recognized, we analyzed this prime candidate gene for genetic variation in carefully selected patients and controls. In this preliminary study, 18 single nucleotide polymorphisms were observed, 17 of which were located in the N-terminal and C terminal region of the coding exon, resulting in 7 amino acid exchanges: Ser-49 Gly, Ala-59-Ser, Gly-389-Arg, Arg-399-Cys, His-402-Arg, Thr-404-Ala, and Pro-418 Ala. These mutations resulted in 11 different beta1-AR genotypes. Importantly, the genotypes carrying the Ser-49-Gly mutation in the N-terminus of the molecule in a heterozygous or homozygous form were observed significantly more frequently in the group of IDCM patients. The present results may provide a clue on the molecular mechanisms involved in IDCM, and add moreover interesting information on nature, distribution, and evolutionary aspects of sequence variation in human adrenergic receptor genes. PMID- 10794545 TI - Differential cell cycle response of nontumorigenic and tumorigenic human papillomavirus-positive keratinocytes towards transforming growth factor-beta1. AB - Human papillomaviruses (HPVs) are causative agents of a number of malignancies in humans, including cervical cancer. Their tumorigenic potential is linked to expression of the viral E6/E7 genes which can interfere with normal cell cycle control by targeting p53, p21WAF1, p27KIP1, and pRb. We show here that nontumorigenic and tumorigenic HPV-positive keratinocytes (HPK) exhibit striking differences in the response of cell cycle regulatory genes towards transforming growth factor beta-beta1. Treatment with this agent led to an efficient induction of p53 and the growth-inhibitory p15INK4 and p21WAF1 genes only in nontumorigenic HPKs and was linked to an efficient reduction in viral E6/E7 oncogene expression. This was associated with increased pRb levels, exhibiting sustained hypophosphorylation, and a permanent growth arrest in the G1 phase of the cell cycle. In contrast, tumorigenic HPKs exhibited only a modest rise in p53 protein levels and a substantially reduced induction of the p15INK4 and p21WAF1 genes, which was linked to a lesser degree of viral oncogene repression. In addition, tumorigenic HPKs rapidly resumed cell growth after a transient G1 arrest, concomitantly with the reappearance of hyperphosphorylated pRb. These results support the notion that the progression of HPV-positive cells to a malignant phenotype is associated with increased resistance to growth inhibition by transforming growth factor-beta1. This is linked in the tumorigenic cells to a lack of persistent G1 arrest, inefficient induction of several cell cycle control genes involved in growth inhibition, and inefficient repression of the growth promoting viral E6/E7 oncogenes. PMID- 10794546 TI - Establishment and characterization of the follicular thyroid carcinoma cell line ML-1. AB - The present study focuses on the establishment and characterization of a new follicular thyroid carcinoma cell line. The human cell line ML-1 was derived from a dedifferentiated follicular thyroid carcinoma relapse, which progressed despite preceding surgery followed by two radioiodine therapies. More than 90% of the cells of this line express thyroglobulin, chondroitin sulfate, and vimentin antigens, but only about 70% show cytokeratin filaments and a negative surface charge density such as human erythrocytes. More importantly, cells of this line are able to take up iodine and/or glucose both in vitro and in vivo and to secrete thyroglobulin, chondroitin sulfate, and fibronectin into the interstitial space. In addition, triiodothyronine is released constitutively into culture supernatants. Moreover, it is also suitable for xenotransplantation studies because it is tumorigenic in NMRI nude mice in vivo. The cell line forms tumors with follicular structures when transplanted to nude mice. Due to these unique features the ML-1 cell line can be considered as a very suitable test model for pharmacological and cell biological studies. Since chemicals may interfere with the production of thyroid hormones, this cell line represents also a tool for toxicological investigations. PMID- 10794547 TI - Increased activity of nm23-H1 gene in squamous cell carcinoma of the head and neck is associated with advanced disease and poor prognosis. AB - The present study was undertaken to determine whether the nm23-H1 gene is expressed in squamous cell carcinoma of the head and neck (SCCHN) and whether the level of nm23-H1 protein or mRNA in cells vary as they progress to a more malignant phenotype. Of the 120 SCCHN studied 54 (45%) stained positively for nm23-H1 protein. Protein expression was significantly higher in more advanced stages of disease. Expression of nm23-H1 was significantly higher in cancer tissues than in normal, adjacent tissue, dysplasia, or carcinoma in situ. The nm23-H1 rate increased with progression of synchronous lesions from dysplasia to carcinoma in situ and finally to carcinoma (P<0.05). Northern blot analyses of tissues with various clinicopathological characteristics also revealed differences in nm23-H1 mRNA expression. When levels of nm23-H1 mRNA were compared to tumor stage, intensity of expression was found to be higher in stages 3 and 4 than stages 1 and 2 (P<0.01). Malignant tumors had a higher level of mRNA nm23-H1 expression than normal or premalignant tissues. The nm23-H1 negative patients survived significantly longer than nm23-H1 positive ones (P<0.05). To study the possible relationship between nm23-H1 gene expression and cell growth rate in tumor cells, the mRNA level in each tumor was compared to proliferative activity. The nm23-H1 gene expression levels were directly related to the [3H]thymidine labeling index in tumor cells (R=0.6681). Our results strongly indicate that the nm23-H1 gene is involved in progression of SCCHN. Together with results obtained on lung cancer, our observations suggest that increased expression of nm23-H1 in cancers of the upper aerodigestive tract may have different implications than elsewhere in the body. PMID- 10794548 TI - Oncogenic osteomalacia. PMID- 10794549 TI - A new classification for cervical vertebral injuries: influence of CT. AB - OBJECTIVE: Computed tomography (CT) has been demonstrated to be superior to radiography in identifying cervical vertebral injuries. However, many of these injuries may not be clinically significant, and require only minimal symptomatic and supportive treatment. It is therefore imperative that radiologists and spine surgeons have criteria for distinguishing between those injuries requiring surgical stabilization and those that do not. The authors propose a new classification of cervical vertebral injuries into two categories: major and minor. DESIGN AND PATIENTS: A data base, acquired on 1052 separate cervical injuries in 879 patients seen between 1983 and 1998, was reviewed. Four categories of injury based on mechanism [hyperflexion (four variants), hyperextension (two variants), rotary (two variants), and axial compression (five variants)] were identified. "Major" injuries are defined as having either radiographic or CT evidence of instability with or without associated localized or central neurologic findings, or have the potential to produce the latter. "Minor" injuries have no radiographic and/or CT evidence of instability, are not associated with neurologic findings, and have no potential to cause the latter. RESULTS AND CONCLUSIONS: Cervical injury should be classified as "major" if the following radiographic and/or CT criteria are present: displacement of more than 2 mm in any plane, wide vertebral body in any plane, wide interspinous/interlaminar space, wide facet joints, disrupted posterior vertebral body line, wide disc space, vertebral burst, locked or perched facets (unilateral or bilateral), "hanged man" fracture of C2, dens fracture, and type III occipital condyle fracture. All other types of fractures may be considered "minor". PMID- 10794550 TI - MR findings of necrotic lesions and the extralesional area of osteonecrosis of the femoral head. AB - OBJECTIVE: To investigate the MR findings of necrotic lesions and the extralesional area of osteonecrosis of the femoral head (ONFH) for each of the radiological stages. DESIGN AND PATIENTS: Forty-nine hips in 29 patients (15 female, 14 male; mean age 38 years, range 17-59 years) were imaged using a 1.0-T superconducting magnet. T2-weighted spin echo pulse sequences (T2WI), spoiled gradient recalled echo pulse sequences (SPGR) and fat suppression SPGR (FS-SPGR), followed by Gd-DTPA enhanced fat suppression SPGR (Gd-FS-SPGR), were all obtained with the aid of a TORSO surface coil. RESULTS AND CONCLUSIONS: While a normal fat intensity area with a low-intensity band on SPGR (band pattern) was seen in 16 of 16 stage 1 (100%), nine of 11 stage 2 (82%), four of 17 stage 3 (24%), and none of five stage 4 hips, all hips showed peripheral rim enhancement on Gd-FS-SPGR (100%). This enhancement band on Gd-FS-SPGR corresponded to histological findings of necrotic trabecular bone, repaired marrow, and fibrous reparative tissue. Bone marrow edema was also clearly demonstrated as a diffuse, high-intensity area outside this enhancement band on Gd-FS-SPGR in two stage 2 (18%), 12 stage 3 (71%), and one stage 4 hip (20%). In cases at stage 2 or more advanced stages with homogeneous or inhomogeneous low intensity on nonenhanced MRI, the reparative process both inside and outside the necrotic lesion, including bone marrow edema, was detected clearly on contrast-enhanced MRI. PMID- 10794551 TI - US and MR imaging of peripheral nerves in leprosy. AB - OBJECTIVE: To analyze peripheral nerves with ultrasonography (US) and magnetic resonance imaging (MR) in leprosy and assess the role of imaging in leprosy patients. DESIGN AND PATIENTS: Fifty-eight nerves with abnormal clinical features or electromyograms were examined in 23 leprosy patients by means of gray-scale US, Doppler US and MR imaging. Image analysis included: measurement of nerve cross-sectional area; assessment of nerve structure and MR signal intensities; identification of nerve compression within osteofibrous tunnels; detection of endoneural color flow signals and Gd-DTPA enhancement. Correlations were made with clinical findings and a control group of 20 subjects. Fourteen nerves in active reversal reaction were followed up after therapy. RESULTS: Leprosy nerves were classified into three groups based on imaging appearance: group I consisted of 17 normal-appearing nerves; group II, of 30 enlarged nerves with fascicular abnormalities; group III, of 11 nerves with absent fascicular structure. Group II nerves were from patients subjected to reversal reactions; 75% of patients with group III nerves had a history of erythema nodosum leprosum. Nerve compression in osteofibrous tunnels was identified in 33% of group II and 18% of group III nerves. Doppler US and MR imaging were 74% and 92% sensitive in identifying active reactions, based on detection of endoneural color flow signals, long T2 and Gd enhancement. In 64% of cases, follow-up studies showed decreased color flow and Gd uptake after steroids and decompressive surgery. CONCLUSIONS: US and MR imaging are able to detect nerves abnormalities in leprosy. Active reversal reactions are indicated by endoneural color flow signals as well as by an increased T2 signal and Gd enhancement. These signs would suggest rapid progression of nerve damage and a poor prognosis unless antireactional treatment is started. PMID- 10794552 TI - Bone density distribution and gender dominate femoral neck fracture risk predictors. AB - OBJECTIVE: To determine whether regional characteristics of the proximal femur could discriminate between a group of patients who had just sustained a first low trauma femoral neck fracture (n=50) from a group of healthy volunteers (n=123). DESIGN: The application of an integral bone measurement (dual-energy X-ray absorptiometry) in conjunction with a volumetric cancellous bone density measurement (quantitative computed tomography) to the proximal femur in vivo provided an estimate of the contribution of the spatial distribution of bone density to hip fracture risk prediction. RESULTS: The primary finding of this study was a significant difference between male and female hip fracture risk predictor variables. In men with femoral neck fracture, a significant decrease in bone density throughout the proximal femur was observed. In women with femoral neck fracture, a combination of local bone deficits (significant decrease in cancellous bone at the site of fracture, and a decrease in cortical bone at the site of impact) and significantly larger proximal femur dimensions (femoral neck and head widths) was evident. CONCLUSIONS: These results imply that effective hip fracture prevention strategies may require separate approaches for men and women. Screening programs for diminished bone density at the proximal femur have proved effective in previous studies. An approach which includes examining these local bone characteristics may further improve our ability to accurately determine hip fracture risk in vivo. PMID- 10794553 TI - Retroisthmic cleft: a stress fracture of the lamina. AB - The retroisthmic cleft is a rarely diagnosed defect in the lamina of the lumbar spine. It has always been considered a congenital anomaly. This is the first report we are aware of showing radiological changes in a retroisthmic cleft over a period of time. The follow-up of this patient over a period of 6 years, the radiological and scintigraphic appearances and a review of the literature suggests that the retroisthmic cleft is a stress fracture of the lamina. PMID- 10794554 TI - Giant distal humeral geode. AB - We describe the imaging features of a giant geode of the distal humerus in a patient with rheumatoid arthritis, which presented initially as a pathological fracture. The value of magnetic resonance imaging in establishing this diagnosis is emphasized. PMID- 10794555 TI - Chondromyxoid fibroma of the acromium with soft tissue extension. AB - Chondromyxoid fibroma is an unusual, benign tumor of cartilaginous origin and represents less than 1% of all primary bone tumors. It usually involves the long bones around the knee joint or the flat bones of the pelvis or ribs. Soft tissue extension is also thought to be rare in these lesions. They are usually eccentrically located in the metaphyses of the long bones and centrally in the flat bones. The radiographic appearances are characteristically those of a single, lytic lesion with lobulated margins, septations, cortical expansion and a sclerotic rim. Histologically, they display a lobulated pattern with spindle shaped cells lying within a myxoid matrix with areas of hyaline cartilage. The differential diagnosis includes giant cell tumor, chondroblastoma or enchondroma as well as chondrosarcoma. The rarity of these lesions may render the diagnosis difficult to make, especially when the lesion involves an unusual site such as the acromium. PMID- 10794557 TI - Metadiaphyseal chondroblastoma of the thumb. AB - Chondroblastoma is typically located in the epiphysis. Predominant metadiaphyseal location is very rare, as is involvement of the digits. We describe a case of chondroblastoma involving the metadiaphysis of the thumb. The patient was a 13 year-old boy who presented with pain and swelling of his left thumb. Radiographs showed an expanded lytic lesion involving the whole metaphysis and diaphysis of the proximal phalanx, which subsequently progressed to involve the epiphysis. Curettage and bone grafting were done. PMID- 10794556 TI - Desmoplastic fibroma of bone: MRI features. AB - Desmoplastic fibroma of bone is a very rare benign tumor, which may be locally aggressive. In contrast to the well-documented radiological appearance, the literature on MR imaging features of this tumor is scarce. The MR imaging characteristics in our case are compared to those previously reported. Although there is a considerable overlap in the MR imaging features with other bone tumors, an interesting MR feature of desmoplastic fibroma is the presence of low to intermediate signal intensity foci on T2-weighted images, which radiographically does not correspond to calcifications. This feature may help narrow the differential diagnosis. PMID- 10794558 TI - Primary angioleiomyoma of the iliac bone: clinical pathological study of one case with flow cytometric DNA content and S-phase fraction analysis. AB - We report on a primary angioleiomyoma of the right iliac bone in a 28-year-old woman. To our knowledge this is the 16th reported case of a primary leiomyoma of bone, the 9th reported primary osseous angioleiomyoma and the first description of a primary angioleiomyoma located in the iliac bone. The problems of differentiating primary leiomyoma of bone from primary or metastatic leiomyosarcoma of bone are stressed. A literature review of primary leiomyomas of bone is presented. PMID- 10794559 TI - A methodology designed to increase accuracy and safety in stereotactic brain surgery. AB - A series of technical tips and devices designed to increase accuracy and safety in stereotactic surgery are presented. We use stereotactic magnetic resonance imaging with three-dimensional magnetization-prepared rapid gradient-echo (MP RAGE) imaging to minimize image distortion, and a three-dimensional stereotactic planning system for accurately registering three-dimensional space. We also developed several technical devices useful for stereotactic intracranial procedures; an applicator system attached to the frame which simulates the fiducial markers in order to keep the target at a suitable position in stereotactic space; a torque wrench to set the torque on the fixing pins to the frame reproducibly at 5 inch pounds in order to keep distortion of the frame to a minimum while maintaining secure fixation; an entry point marker to maintain the calculated trajectory angle; a straightening cannula to prevent the thermo coagulation needle from bending; a microvascular Doppler and its holder to detect significant vessels and to know their precise depth in order to avoid vascular injury from thermocoagulation; a burr hole button device to secure depth electrode cables at the patient's skull. PMID- 10794560 TI - Biportal endoscopic removal of a primary intraventricular hematoma: case report. AB - Primary intraventricular hematomas account for approximately 6% of all intracerebral hematomas. If the clot blocks cerebrospinal fluid (CSF) pathways, surgical intervention, which may be of different types, can be life-saving. In the case reported here, after careful preoperative planning the use of two rigid endoscopes permitted the removal of most of the intraventricular clot and restoration of CSF circulation by creation of a 3rd ventriculostomy within the same procedure and no later treatment was necessary. Repeated CT scans proved that only a small portion of the intraventricular clot remained in the ventricular system. The ventricular size normalised, and the patency of the artificial hole in the floor of the 3rd ventricle was demonstrated both by the rapidly improving clinical picture of the patient and by flow-sensitive MRI studies. For individuals who suffer primary intraventricular hemorrhage and later develop occlusive hydrocephalus, endoscopic removal of the clot and 3rd ventriculostomy might offer a more adequate treatment option than external ventricular drainage. PMID- 10794561 TI - Frameless stereotactically guided catheter placement and fibrinolytic therapy for spontaneous intracerebral hematomas: technical aspects and initial clinical results. AB - OBJECTIVE: Frame-based stereotactic puncture and catheter placement followed by fibrinolytic therapy and drainage is one treatment option in the management of spontaneous intracerebral hemorrhage (sICH). This minimally invasive procedure could even be simplified by frameless stereotaxy. The authors present their experiences with frameless stereotactic image-guided catheter placement for lysis and drainage of sICH, with emphasis on technical aspects. METHOD: In 27 patients with sICH, an infrared-based frameless stereotactic device was used for selecting trajectory and target point of hematoma drainage. A trajectory along the main axis of the hematoma was considered to be optimal for fibrinolytic therapy. An articulated arm served to maintain the predetermined trajectory during surgery and to guide catheter advancement. Clot lysis with recombinant tissue plasminogen activator (rt-PA) was initiated after radiological confirmation of correct catheter positioning. RESULTS: In all cases, selection of the optimal trajectory was not restricted by the frameless stereotactic device. In 25 of the 27 patients, the catheter was placed accurately along the predetermined trajectory into the target point. In two patients, the catheter was positioned at the lateral margin of the hematoma, excluding fibrinolytic therapy in one case. In 24 of 27 patients, the mean hematoma volume could be reduced from initially 52 ml to 17 ml in an average of two days. Hematoma enlargement following rt-PA injection was observed in two patients. Further complications were culture negative pleocytosis of cerebrospinal fluid in two and meningitis in one patient. CONCLUSION: Hematoma puncture and catheter placement for fibrinolytic therapy could be achieved with high accuracy and safety using frameless stereotaxy. This method allows unrestricted trajectory selection with catheter positioning along the main hematoma axis. Further studies are required to investigate if frameless stereotactic puncture and clot lysis could contribute to improve the outcome of patients with sICH. PMID- 10794562 TI - Preoperative embolization of intracranial meningiomas: a 17-years single center experience. AB - The purpose of the present report is to review the evolution of endovascular therapy at our center as utilized for the preoperative embolization of intracranial meningiomas over a 17-years period (1982-1998). This study is based upon a consecutive series of 63 patients who underwent preoperative embolization of intracranial meningiomas. Total or subtotal angiographic devascularization of the tumor parenchyma was accomplished in 38 patients (60.3%) who had tumors with either an external carotid artery supply only (n = 30) or with contributions from the cavernous carotid artery, ophthalmic artery, vertebral artery, or pial feeders which were feasible for selective embolization (n = 8). Partial tumor embolizations were attained in the remaining 25 patients (39.7%) because 1. the remanent feeders were considered easily accessible to surgical control in the early stages of dissection, 2. the feeding branches were inaccessible for a microcatheter approach, or 3. superselective microcatheter positions allowing for safe embolization without reflux of embolic material into physiological branches were not achieved. Overall, 97 of 126 tumor feeders identified angiographically were catheterized to selective embolization (77%). Three embolization related complications occurred early in our experience (1982-1989) using techniques which no longer meet standards of treatment. In light of the remarkable evolution of endovascular techniques over the 17-years study period, however, we conclude that preoperative embolization of intracranial meningiomas can be performed safely with the endovascular tools currently available. PMID- 10794563 TI - Successful treatment of large malignant tumor involving the skull base by radiosurgery combined with intraarterial chemotherapy and embolization. AB - Stereotactic radiosurgery can be used to treat malignant tumors involving the skull base. While it avoids extensive sacrifice of the normal structures surrounding the tumor, radiosurgery does not eradicate the tumor mass immediately. We present a patient with a large hypervascular malignant tumor involving the skull base that resulted in intracranial hypertension. He was successfully treated with stereotactic radiosurgery combined with intra-arterial chemotherapy and embolization of the arteries feeding the tumor. We discuss radiosurgery, chemotherapy, embolization and other therapeutic modalities for treating large malignant tumors involving the skull base. PMID- 10794564 TI - Transsphenoidal supradiaphragmatic intradural approach: technical note. AB - Presellar extension of the bone window combined with removal of the sellar floor results in the transsphenoidal supradiaphragmatic intradural approach. One tuberculum sella meningioma and another suprasellar Rathke's cleft cyst confined to the pituitary stalk were removed via this approach. The presellar extension of the bone window was performed with the sublabial transseptal transsphenoidal technique. Furthermore, the dissection of the anterior intercavernous sinus, diaphragma sella, and arachnoid trabecula has allowed a wide surgical field of pre- and suprasellar areas and facilitates safe removal of lesions without significant surgical complications in selected cases. PMID- 10794565 TI - Endoscopic endonasal transsphenoidal surgery in recurrent and residual pituitary adenomas: technical note. AB - Despite a good cure rate after surgery, the recurrence rate in pituitary adenomas is globally high. The decision making in such cases can be problematic for the nature of the lesion, for the anatomic structures involved, for the different pharmacological, surgical, radiotherapeutic and radiosurgical options nowadays available. In the perspective of an improvement and refinement of the surgical procedure an endoscopic endonasal transsphenoidal approach to the pituitary was recently adopted in the Neurosurgical Department of the University of Naples. Its minimal invasiveness and its wider and direct anatomic control of the operative field has allowed a faster, greater and safer potential of tumour excision, with respect of the sphenoid, sellar and parasellar structures. The authors have examined the advantages provided by this technique in 12 patients with recurrent pituitary adenomas and in 2 craniopharyngiomas already treated via a transnasal transsphenoidal approach (TTA), where the anatomy of surgical field had been distorted by the first operation or the radiation therapy. They conclude that the endoscopic transsphenoidal re-operation might be considered the procedure of choice in case of recurrences and its easiness in such conditions could favour its larger use, before other more aggressive therapeutic solutions. PMID- 10794566 TI - Augmentative treatment of chronic deafferentation pain syndromes after peripheral nerve lesions. AB - Deafferentation pain syndromes developing after peripheral nerve lesions are difficult to treat. According to the follow-up (mean: 39.5 months) of 6 patients suffering from causalgic pain we will present our method of augmentative therapy in chronic neuropathic pain caused by peripheral nerve lesions, i.e., peripheral nerve stimulation (PNS), spinal cord stimulation (SCS) and chronic intrathecal opioid infusion. None of the patients showed intraoperative or follow-up complications. Evaluated by visual analogue scales all patients reported a good to excellent pain relief (75-100%). (1) Regarding the favourable long-term results of PNS, this method should be considered in cases of mononeuropathic pain syndromes. (2) Neuropathic pain syndromes which are not assignable to a singular nerve lesion, can often be managed effectively by SCS. (3) In contrast to the widespread opinion, deafferentation pain syndromes of central or peripheral origin can be treated satisfactorily by intrathecal opiate administration. PMID- 10794567 TI - Multiple oligodendroglioma: case report. AB - An 18-year-old female patient was hospitalized with headache and disturbance of consciousness. Magnetic resonance imaging (MRI) revealed a tumor in the left parieto-occipital lobe. The tumor was totally removed, and postoperative radiation therapy was administered locally at 50 Gy. Ten months later, she experienced sudden onset of unconsciousness and headache. Computed tomography (CT) and MRI demonstrated multiple mass lesions in the whole brain. Following the systemic chemotherapy, removal of the largest tumor was performed. Histological examination proved all excised tumors to be oligodendroglioma without evidence of malignant change. PMID- 10794568 TI - Neuroendoscopic biopsy of tectal glioma: a case report. AB - OBJECTIVE AND IMPORTANCE: A type of intrinsic dorsal midbrain glioma known as benign tectal glioma can cause obstructive hydrocephalus. Because of its slow progress and relatively good prognosis, initial treatment should be cerebrospinal fluid diversion and biopsy. CLINICAL PRESENTATION: We report a 24-year-old man with obstructive hydrocephalus from a tectal glioma, who was admitted to the hospital for malfunction of a shunt placed 16 years previously. INTERVENTION: Magnetic resonance imaging demonstrated enlargement of the dorsal midbrain associated with increased signal intensity on T2-weighted images and fluid attenuation inversion recovery (FLAIR) images. No enhancement occurred with contrast administration. We performed a neuroendoscopic third ventriculostomy and biopsy of the tumor. Histologic examination of the specimen obtained disclosed a low-grade astrocytoma. No postoperative neurologic complications occurred. CONCLUSION: FLAIR images were superior to T2-weighted images in demonstrating abnormal intensity in the dorsal midbrain. Neuroendoscopy confirmed a yellowish tumor inferior to the posterior commissure occluding the entrance of the aqueduct, an observation useful in planning biopsy of the lesion. As the tumor apparently had existed for a number of years, the case supports the notion that the prognosis associated with these tumors is good. PMID- 10794569 TI - Heterogeneity of eukaryotic replicons, replicon clusters, and replication foci. AB - According to the current paradigm, replication foci are discrete sites in the interphase nucleus where assemblies of DNA replication enzymes simultaneously elongate the replication forks of 10-100 adjacent replicons (each approximately 100 kbp). Here we review new results and provide alternative interpretations for old results to show that the current paradigm is in need of further development. In particular, many replicons are larger than previously thought - so large that their complete replication takes much longer (several hours) than the measured average time to complete replication at individual foci (45-60 min). In addition to this large heterogeneity in replicon size, it is now apparent that there is also a corresponding heterogeneity in the size and intensity of individual replication foci. An important property of all replication foci is that they are stable structures that persist, with constant dimensions, during all cell cycle stages including mitosis, and therefore likely represent a fundamental unit of chromatin organization. With this in mind, we present a modified model of replication foci in which many of the foci are composed of clusters of small replicons as previously proposed, but the size and number of replicons per focus is extremely heterogeneous, and a significant proportion of foci are composed of single large replicons. We further speculate that very large replicons may extend over two or more individual foci and that this organization may be important in regulating the replication of such large replicons as the cell proceeds through S phase. PMID- 10794570 TI - Immunofluorescent studies of human chromosomes with antibodies against phosphorylated H1 histone. AB - One of the prominent cell cycle-related modifications of histone proteins whose function remains unresolved is the phosphorylation of linker histone H1. In this work we have used indirect immunofluorescence on human cells with antibodies that are specific for phosphorylated histone H1 to examine the cellular distribution and chromosome association patterns of this protein. With confocal microscopy on whole cells, strong immunofluorescence was seen in association with mitotic chromosomes as well as a prominent punctate pattern of labeling throughout the mitotic cell, whereas interphase cells showed very little, if any, specific fluorescence. Multiple patterns of fluorescence distribution were detected with metaphase chromosomes, ranging from apparent tight colocalization with the DNA to expanded "puffy" mitotic figures to an amorphous network of staining. It was also shown that the ability to label chromosomes could vary drastically with different fixation procedures, adding further complications to interpretation of the potentially complex role of phosphorylated histone H1 in chromatin condensation or decondensation. PMID- 10794572 TI - Architecture of the Trypanosoma brucei nucleus during interphase and mitosis. AB - The structural basis of mitosis, spindle organisation and chromosome segregation, in the unicellular parasite Trypanosoma brucei is poorly understood. Here, using immunocytochemistry, fluorescent in situ hybridisation and electron microscopy, we provide a detailed analysis of mitosis in this parasite. We describe the organisation of the mitotic spindle during different stages of mitosis, the complex ultrastructure of kinetochores and the identification of a potential spindle-organising centre in the mitotic nucleus. We investigate the dynamics of chromosome segregation using telomeric and chromosome-specific probes. We also discuss the problems involved in chromosome segregation in the light of the fact that the T. brucei karyotype has 22 chromosomes in the apparent presence of only eight ultrastructurally defined kinetochores. PMID- 10794571 TI - Unusual core histones specifically expressed in male gametic cells of Lilium longiflorum. AB - We have cloned three novel histone genes using antibodies that recognize only nuclei of the male gametic (generative and sperm) cells of Lilium longiflorum. The deduced amino acid sequence of each clone shows only between 40% and 50% identity with the H2A, H2B and H3 somatic core histones of other plant species. Transcripts of these genes were first detected in bicellular pollen soon after microspore mitosis, and their mRNAs, as revealed by in situ hybridization, were observed only in the cytoplasm of the generative cells. As expression of these three genes was specific to generative cells within the bicellular pollen, we designated the clones gH2A, gH2B and gH3. Immunocytochemistry further revealed that the proteins encoded by these genes accumulated in the elongating and condensing generative nucleus during development of bicellular pollen, and were most abundant in the two sperm nuclei within an elongated pollen tube. We therefore propose that these male gamete-specific core histones contribute to chromatin condensation of male gametes or to chromatin remodeling, and result in the repression of gene expression in male gametes. PMID- 10794573 TI - Intranuclear arrangement of human chromosome 12 correlates to large-scale replication domains. AB - The intranuclear arrangement of human chromosome 12 in G0(G1) nuclei from human myeloid leukemia HL60 cells was analyzed by multicolor fluorescence in situ hybridization (FISH) using band-specific cosmid clones as probes. Pairs of differently colored cosmids were detected on paraformaldehyde-fixed HL60 nuclei, and their relative positions, internal or peripheral, in individual nuclei were scored. Our results suggest that the intranuclear arrangement of human chromosome 12 is not random. Some chromosomal domains, including the centromere, were located in the periphery of the nucleus, while other domains, including the telomeres, were positioned in the internal areas of the nucleus in GO(G1) cells. Based on the replication banding patterns of metaphase spreads, human chromosome 12 was divided roughly into five large domains. Interestingly, the clones in late replicating domains were preferentially localized in the nuclear periphery, whereas clones in early replicating domains were arranged in the internal areas of the nuclei. The DNA replication timing of each cosmid determined by FISH-based assay did not reflect the replication bands, but an overall profile of the replication timing was relatively correlated with these domains on chromosome 12. These results suggest that the intranuclear arrangement of a human chromosome is correlated with the large-scale replication domains, even before DNA replication. PMID- 10794574 TI - Evolution of different subfamilies of mariner elements within the medfly genome inferred from abundance and chromosomal distribution. AB - The abundance and distribution pattern of eight mariner elements from three different subfamilies in the genome of the medfly Ceratitis capitata were determined. The copy numbers, as determined by slot-blot analysis, were very different for these elements. Their abundance did not change significantly within the native, the ancient or the newly derived populations, indicating that the rapid colonization process of the medfly had not affected the copy number of mariner elements. The distribution of the mariner elements was analyzed using fluorescent in situ hybridization (FISH) with charge-coupled device (CCD) camera analysis. The pattern of distribution in euchromatin and heterochromatin varied greatly and was distinctive and specific for each element. The implications of these findings are discussed and it is concluded that they generally support the hypothesis of a transposition/selection model in which the abundance and distribution patterns of these elements are regulated primarily by selection against deleterious effects due to meiotic ectopic recombination, while genetic drift would have played a minor role. PMID- 10794575 TI - Evaluation of common breast problems in family practice. PMID- 10794576 TI - Improving adherence to asthma therapy: what physicians can do. PMID- 10794577 TI - Hiding consumer ads in pharmaceutical samples. PMID- 10794578 TI - Use of intravenous colchicine in patients with acute gout. PMID- 10794579 TI - The evaluation of common breast problems. AB - The most common breast problems for which women consult a physician are breast pain, nipple discharge and a palpable mass. Most women with these complaints have benign breast disease. Breast pain alone is rarely a presenting symptom of cancer, and imaging studies should be reserved for use in women who fall within usual screening guidelines. A nipple discharge can be characterized as physiologic or pathologic based on the findings of the history and physical examination. A pathologic discharge is an indication for terminal duct excision. A dominant breast mass requires histologic diagnosis. A breast cyst can be diagnosed and treated by aspiration. The management of a solid mass depends on the degree of clinical suspicion and the patient's age. PMID- 10794580 TI - Acute knee effusions: a systematic approach to diagnosis. AB - Knee effusions may be the result of trauma, overuse or systemic disease. An understanding of knee pathoanatomy is an invaluable part of making the correct diagnosis and formulating a treatment plan. Taking a thorough medical history is the key component of the evaluation. The most common traumatic causes of knee effusion are ligamentous, osseous and meniscal injuries, and overuse syndromes. Atraumatic etiologies include arthritis, infection, crystal deposition and tumor. It is essential to compare the affected knee with the unaffected knee. Systematic physical examination of the knee, using specific maneuvers, and the appropriate use of diagnostic imaging studies and arthrocentesis establish the correct diagnosis and treatment. PMID- 10794581 TI - Addiction: part II. Identification and management of the drug-seeking patient. AB - The medications most often implicated in prescription drug abuse are opioid analgesics, sedative-hypnotics and stimulants. Patients with acute or chronic pain, anxiety disorders and attention-deficit disorder are at increased risk of addiction comorbidity. It is important to ask patients about their substance-use history, including alcohol, illicit drugs and prescription drugs. Patients who abuse prescription drugs may exhibit certain patterns, such as escalating use, drug-seeking behavior and doctor shopping. A basic clinical survival skill in situations in which patients exert pressure on the physician to obtain a prescription drug is to say "no" and stick with it. Physicians who overprescribe can be characterized by the four "Ds"-dated, duped, dishonest and disabled. Maintaining a current knowledge base, documenting the decisions that guide the treatment process and seeking consultation are important risk-management strategies that improve clinical care and outcomes. PMID- 10794582 TI - Acute otitis media: part II. Treatment in an era of increasing antibiotic resistance. AB - Antibiotic resistance is increasing among the pathogens that commonly cause acute otitis media. This development may merit changes in the traditional antibiotic treatment of acute otitis media. Risk factors for resistant pathogens include recent antibiotic treatment of acute otitis media, children in day care facilities, wintertime infections and acute otitis media in children less than two years of age. Amoxicillin remains the antibiotic of first choice, although a higher dosage (80 mg per kg per day) may be indicated to ensure eradication of resistant Streptococcus pneumoniae. Oral cefuroxime or amoxicillin-clavulanate and intramuscular ceftriaxone are suggested second-line choices for treatment failure. Compliance with antibiotic regimens is enhanced by selecting agents that require less frequent dosing (such as one or two times a day) and by prescribing shorter (five days or less) treatment courses. Selective use of tympanocentesis if the patient does not respond to empiric therapy can help confirm the diagnosis and guide effective therapy. PMID- 10794583 TI - Developing and communicating a long-term treatment plan for asthma. AB - The treatment of asthma, according to current guidelines, requires complex treatment regimens that change as clinical conditions improve or deteriorate. We have developed a practical way to communicate long-term treatment plans in chart form in the primary care setting that is easy for patients to follow and use. The chart has been an important element in two interventions that have resulted in positive changes in health behavior and health outcomes in children with asthma. The plan provides recommendations for patients and families to make adjustments in medication based on changes in symptoms or peak expiratory air flow, or both, that are consistent with the Asthma Guidelines Expert Panel Report 2, 1997. The plan also indicates when the number and dosage of drugs should be increased or decreased and when emergency care should be sought, consistent with the Asthma Guidelines. By placing considerable control in the family's hands and by clearly delineating the conditions under which medicines can be reduced or discontinued, the physician provides incentives for families to adhere to the long-term treatment plan for asthma. PMID- 10794584 TI - Management of herpes zoster (shingles) and postherpetic neuralgia. AB - Herpes zoster (commonly referred to as "shingles") and postherpetic neuralgia result from reactivation of the varicella-zoster virus acquired during the primary varicella infection, or chickenpox. Whereas varicella is generally a disease of childhood, herpes zoster and post-herpetic neuralgia become more common with increasing age. Factors that decrease immune function, such as human immunodeficiency virus infection, chemotherapy, malignancies and chronic corticosteroid use, may also increase the risk of developing herpes zoster. Reactivation of latent varicella-zoster virus from dorsal root ganglia is responsible for the classic dermatomal rash and pain that occur with herpes zoster. Burning pain typically precedes the rash by several days and can persist for several months after the rash resolves. With postherpetic neuralgia, a complication of herpes zoster, pain may persist well after resolution of the rash and can be highly debilitating. Herpes zoster is usually treated with orally administered acyclovir. Other antiviral medications include famciclovir and valacyclovir. The antiviral medications are most effective when started within 72 hours after the onset of the rash. The addition of an orally administered corticosteroid can provide modest benefits in reducing the pain of herpes zoster and the incidence of postherpetic neuralgia. Ocular involvement in herpes zoster can lead to rare but serious complications and generally merits referral to an ophthalmologist. Patients with postherpetic neuralgia may require narcotics for adequate pain control. Tricyclic antidepressants or anticonvulsants, often given in low dosages, may help to control neuropathic pain. Capsaicin, lidocaine patches and nerve blocks can also be used in selected patients. PMID- 10794585 TI - Evaluation and treatment of swallowing impairments. AB - Swallowing disorders are common, especially in the elderly, and may cause dehydration, weight loss, aspiration pneumonia and airway obstruction. These disorders may affect the oral preparatory, oral propulsive, pharyngeal and/or esophageal phases of swallowing. Impaired swallowing, or dysphagia, may occur because of a wide variety of structural or functional conditions, including stroke, cancer, neurologic disease and gastroesophageal reflux disease. A thorough history and a careful physical examination are important in the diagnosis and treatment of swallowing disorders. The physical examination should include the neck, mouth, oropharynx and larynx, and a neurologic examination should also be performed. Supplemental studies are usually required. A videofluorographic swallowing study is particularly useful for identifying the pathophysiology of a swallowing disorder and for empirically testing therapeutic and compensatory techniques. Manometry and endoscopy may also be necessary. Disorders of oral and pharyngeal swallowing are usually amenable to rehabilitative measures, which may include dietary modification and training in specific swallowing techniques. Surgery is rarely indicated. In patients with severe disorders, it may be necessary to bypass the oral cavity and pharynx entirely and provide enteral or parenteral nutrition. PMID- 10794586 TI - Significant FDA approvals in 1999. PMID- 10794587 TI - AHA and ACC outline approaches to coronary disease risk assessment. PMID- 10794588 TI - The role of iron in Parkinson disease and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine toxicity. AB - Parkinson disease (PD) involves the specific degeneration of dopaminergic neurons of the pars compacta of the substantia nigra. Although the cause of the degeneration of nigrostriatal dopaminergic neurons in PD is unknown, there is significant evidence to suggest that oxidative stress may be involved in this process. This review specifically examines the current status of evidence suggesting iron may contribute to oxidative damage associated with PD. PMID- 10794589 TI - Caspases in developmental cell death. AB - Caspases are a family of evolutionarily conserved cysteine proteases that constitute the effector arm of the apoptotic machinery. Studies in Caenorhabditis elegans, Drosophila melanogaster, and mouse point to evolutionarily conserved caspase function in developmentally programmed cell death in metazoans. Whereas in the nematode all developmental cell death is mediated by a single caspase, in Drosophila and the mouse some caspases appear to regulate cell death in a spatio temporally restricted manner. This article reviews what we currently know about the roles of various caspases in the execution of developmentally programmed cell death and what may be expected from future research in this field. PMID- 10794590 TI - Evolutionary conservation of the membrane fusion machine. AB - Recent structural studies of proteins mediating membrane fusion reveal intriguing similarities between diverse viral and mammalian systems. Particularly striking is the close similarity between the transmembrane envelope glycoproteins from the retrovirus HTLV-1 and the filovirus Ebola. These similarities suggest similar mechanisms of membrane fusion. The model that fits most currently available data suggests fusion activation in viral systems is driven by a symmetrical conformational change triggered by an activation event such as receptor binding or a pH change. The mammalian vesicle fusion mediated by the SNARE protein complex most likely occurs by a similar mechanism but without symmetry constraints. PMID- 10794591 TI - Superoxide and iron: partners in crime. AB - Superoxide (O2-) poses multiple threats, which are diminished by a family of metalloenzymes, the superoxide dismutases. Among the damaging effects of O2- are direct oxidation of low-molecular-weight reductants; inactivation of a select group of enzymes; and reaction with NO to yield the strong oxidant, peroxynitrite. Of even greater import is the ability of O2- to univalently oxidize the [4 Fe-4 S] clusters of dehydratases, which causes release of iron. The "free" iron, which is kept reduced by cellular reductants, then reduces hydroperoxides to hydroxyl or alkoxyl radicals. Because the "free" iron will preferentially bind to anionic polymers, such as nucleic acids, or to anionic surfaces, such as cell membranes, these radicals will be generated adjacent to these vital targets and will preferentially attack them. O2- and iron can thus be viewed as partners in crime, and reciprocal regulatory effects between iron and O2- may be anticipated. These are discussed. PMID- 10794592 TI - DNA-dependent RNA polymerase II from Candida species is a multiple zinc containing metalloenzyme. AB - We have purified DNA-dependent RNA polymerase II from Candida albicans, a human pathogenic yeast. The enzyme consists of 9 polypeptides that are unique to C. albicans, their mobility on SDS-PAGE being different from the mobility of the corresponding subunits of RNA polymerase II from Saccharomyces cerevisiae or C. utilis. In the present study we also demonstrate that RNA pol II from C. albican and C. utilis are metalloproteins containing approximately 5 mol of zinc per mole of enzyme. Although prolonged dialysis in 10 or 20 mM EDTA failed to remove Zn(II) from the C. albicans enzyme, in the C. utilis enzyme 3 Zn(II) ions could be removed and then reconstituted in the presence of excess Zn(II). o Phenanthroline (5 mM) removed Zn(II) from C. albicans enzyme irreversibly in a time-dependent fashion with concomitant loss of enzyme activity. Circular dichroism studies revealed structural changes on removal of zinc, thus suggesting a role for Zn in maintenance of structural stability. Further, we demonstrate that the largest subunit of the C. utilis enzyme and the 3 large subunits of the C. albicans enzyme can bind radioactive zinc. PMID- 10794594 TI - Use of novel bone biopsy system to study molecular effects of growth hormone in human bone--a pilot study. AB - In this study, we have examined whether a novel bone biopsy system combined with reverse transcription-polymerase chain reaction (RT-PCR) or differential display PCR (ddPCR) can be used to detect specific mRNAs induced by growth hormone (GH) in human bone. In a 58-year-old man with complete GH deficiency as a result of empty sella, bone biopsies were taken before, and 5 and 24 h after administration of 24 recombinant human GH. Insulin-like growth factor binding protein-3 (IGFBP 3) mRNA levels in this patient, measured in a semiquantitative RT-PCR assay, increased about 40% 24 h after GH administration. This increase was not seen in a healthy control who did not receive GH, suggesting that the increase was an effect of GH rather than of the biopsy itself. Several differentially expressed mRNAs were detected by ddPCR. Thus, this pilot study suggests that our novel bone biopsy system may be suitable for in vivo studies of the molecular effects of substances with essential functions in human bone. PMID- 10794593 TI - Expression of a human serum albumin fragment (consisting of subdomains IA, IB, and IIA) and a study of its properties. AB - Site-directed mutagenesis and a yeast expression system were used to synthesize a human serum albumin (HSA) fragment (amino acids 1-297). The HSA fragment (half HSA) was evaluated with a number of biophysical techniques and found to be similar to the corresponding region in wild-type HSA. Specifically, the circular dichroism spectra of half HSA and wild-type HSA were superimposable, indicating that the highly alpha-helical secondary structure of wild-type HSA is preserved in half HSA. Additionally, half HSA was partially reactive with a polyclonal antibody against authentic HSA. Half HSA, which contains subdomain IIA, had an affinity for thyroxine and several thyroxine analogs, similar to that observed previously for wild-type HSA. This study suggests that the production of recombinant HSA fragments will be useful for the study of HSA ligand interactions. PMID- 10794595 TI - Induction of neutrophil calcium mobilization by phosphatidic acid-enriched plasma membranes. AB - The purpose of this investigation was to determine if phosphatidic acid (PA) confined to biological membranes could induce physiological responses similar to those induced by exogenous PA. Plasma membranes were treated with phospholipase D (PLD) to increase concentrations of PA within the membranes. Membranes were also treated with other phospholipases including phospholipase A2 (PLA2), and phospholipase C (PLC), which degrade phospholipids without generating PA. A phosphatidylinositol (PI) 3'-kinase inhibitor, LY294002, strongly and selectively inhibited intracellular calcium mobilization induced by PLD-treated membranes. This study suggests that PA-enriched plasma membranes, which exert their effects by activating a unique signaling pathway mediated by PI 3'-kinase, are potent, physiologically relevant initiators of neutrophil activation. PMID- 10794596 TI - Allosteric properties of cyanobacterial cytochrome c oxidase: inhibition of the coupled enzyme by ATP and stimulation by ADP. AB - Thorough analysis of the cta operon of Synechocystis sp. PCC6803 (grown in high concentration salt medium to enhance the expression of respiratory proteins) showed that, apart from ctaCDE and Fb genes potentially encoding subunits I, II, III, and a small pseudo-bacteria-like subunit-IV of unknown function, a large mitochondria-like cta-Fm gene and a pronounced terminator structure are additional components of the operon. The deduced cta Fm gene product shows approximately 50% and 20% sequence identity to the Saccharomyces cerevisiae and beef heart mitochondrial COIV proteins, respectively. It also shows amino acid regions (near the N terminus, on the cytosolic side) with conspicuous sequence similarities to adenylate-binding proteins such as ATP synthase beta subunit Walker A and B consensus regions or to adenylate kinase. We suggest that, similar to the situation with beef heart mitochondria, it is the mitochondria-like subunit-IV of the cyanobacterial aa3-type cytochrome-c oxidase that confers allosteric properties to the cyanobacterial enzyme, the H+/e- ratios of cytochrome c oxidation being significantly lowered by ATP (intravesicular or intraliposomal) but enhanced by ADP. Therefore, the antagonistic action of ATP and ADP was in a way that the redox reaction proper, was always significantly less affected than the coupled proton translocation. Evolutionary and ecological implications of the unusual allosteric regulation of a prokaryotic cytochrome-c oxidase is discussed. PMID- 10794597 TI - Site-directed mutagenesis of Asp313, Glu315, and Asp391 residues in chitinase of Aeromonas caviae. AB - Site-directed mutagenesis was used to explore the roles of amino acid residues involved in the activity of chitinase from Aeromonas caviae. Kinetic parameters for 4-methylumbelliferyl-N,N'-diacetyl-chitobiose or 4-methylumbelliferyl-N,N',N" triacetylchitotriose hydrolysis were determined with wild-type and mutant chitinases. Chitinases with the mutations E315D (or Q) and D391E (or N) were severely impaired and had dramatically decreased kcat. However, the effect of the these mutations on the Km values were different. The function of the carboxyl group of Asp313 was partially replaced by the amide of Asn when the 4 methylumbelliferyl-N,N',N"-triacetylchitotriose substrate was used. Results indicated that Asp313, Glu315, and Asp391 might be the best candidates for the catalytic residues of chitinase A from Aeromonas caviae. PMID- 10794598 TI - Association between angiotensin-converting enzyme gene polymorphism and coronary artery disease. AB - An insertion/deletion (I/D) polymorphism in the gene for angiotensin-converting enzyme (ACE) is associated with myocardial infarction and other cardiac pathology. There is evidence for a role of the renin-angiotensin system in cell growth and in the repair of damaged arterial walls, so the ACE gene is postulated to be a candidate gene affecting the important clinical problem of coronary artery disease (CAD). In view of the clinical importance of the ACE as a major marker of cardiovascular diseases, we investigated the I/D polymorphism of the ACE gene in Turkish CAD patients in comparison with control subjects to evaluate a possible association between CAD and the gene encoding ACE. Polymerase chain reaction, restriction fragment length polymorphism, and agarose gel electrophoresis techniques were used to determine the ACE genotype in 58 subjects. The frequencies of ACE D and ACE I allele among the patients with CAD were 62.26% and 37.73 % and in the control subjects were 49.3% and 50.76%, respectively. The greater frequency of deletion allele (D) was in the CAD group than in the control subjects was significant (P < 0.01). PMID- 10794599 TI - Isolation and characterization of a mutant defective in the production of methanol dehydrogenase from a new restricted facultative methanol-oxidizing bacterium. AB - A new restricted facultative methanol-oxidizing bacterium Methylobacillus sp. strain SK1 (DSM 8269) was subjected to insertion mutagenesis studies with the transposon Tn5 to generate mutants deficient in the production of methanol dehydrogenase (MDH). The transposon was conjugally transferred into the methylotroph by using the triparental mating procedure. The mutants induced by Tn5 were selected directly from a plate containing succinate by using the allyl alcohol selection procedure. The transposition of Tn5 to the bacterium showed a moderate transposition frequency (10(-5) - 10(-6)). Southern hybridization analysis confirmed that the transposon Tn5 was inserted into the chromosomal DNA of the mutants. In addition, the mutants had no RNA transcripts produced from the mdh gene, as judged by Northern blot analysis. The mutants could neither grow on methanol nor produce MDH protein, as determined by Western blot analysis with anti-MDH antibody. These results suggest that Tn5 mutagenesis maybe a useful tool for the molecular analysis of a facultative methanol-oxidizing bacterium. PMID- 10794600 TI - Involvement of ArcA and Fnr in expression of Escherichia coli thiol peroxidase gene. AB - To explore the oxygen response regulators involved in thiol peroxidase gene (tpx) expression in Escherichia coli, we constructed a single-copy tpx-lacZ operon fusion and monitored tpx-lacZ expression in various genetic backgrounds. Expression of the tpx-lacZ fusion was increased 4-fold by aerobic growth. Anaerobic expression of tpx-lacZ in either (delta)arcA or delta(fnr) strains was 2.5-fold depressed compared with that of the wild-type strain. The results of immunoblotting experiments also demonstrated that ArcA and Fnr regulatory proteins repressed thiol peroxidase gene expression during anaerobic growth. Inspection of the tpx promoter region revealed putative binding sites for ArcA and Fnr. It thus appears that ArcA and Fnr function as repressors by blocking the binding of RNA polymerase to the tpx promoter in E. coli under anaerobic growth conditions. PMID- 10794601 TI - New chemiluminescence assay for linamarin. AB - A new chemiluminescence assay was developed for the quantitative determination of linamarin, a cyanogenic glucoside present in cassava. The assay involved hydrolysis of linamarin by a specific enzyme, linamarase, to release glucose, which was then quantitated by a chemiluminescence system consisting of glucose oxidase-peroxidase-luminol. The new assay was more sensitive than the conventional spectrophotometric method for quantitating linamarin in cassava extracts. However, the following agents were found to interfere with the new assay: Vanadate, Mg2+, and Cu2+, were inhibitory to the luminescence of the H2O2 peroxidase-luminol system used in the coupling reaction, whereas EDTA and EGTA activated the system. In addition, Hg2+, which inhibits glucose oxidase, and Tris ion, which inhibits linamarase, both interfered with the new assay. PMID- 10794602 TI - Phosphorylation-dephosphorylation of retinoblastoma protein not necessary for passage through the mammalian cell division cycle. AB - Phosphorylation of the retinoblastoma protein (Rb) during the G1-phase of the mammalian cell division cycle is currently believed to be a controlling element regulating the passage of cells into S-phase. We find, however, that the suspension-grown cell lines U937, L1210, and MOLT-4 contain exclusively hyperphosphorylated Rb. Furthermore, when adherent NIH3T3 cells are grown at very low densities to avoid overgrowth and contact inhibition, they also contain only hyperphosphorylated Rb. NIH3T3 cells exhibit hypophosphorylation when the cells are grown at moderate to high cell densities. We propose that cultures of adherent cells such as NIH3T3, when grown to moderate cell densities, are made up of two populations of cells: (a) cells that are relatively isolated and therefore growing exponentially without contact inhibition, and (b) cells that are growth inhibited by local cell density or contact inhibition. The common observation in adherent cell lines, that Rb is both hyper- and hypophosphorylated in the G1 phase and only hyperphosphorylated in the S- and G2-phases, is explained by the effects of cell density and contact inhibition. Thus, phosphorylation dephosphorylation of Rb protein during the G1 phase is not a necessary process during the NIH3T3, L1210, MOLT-4, and U937 division cycles. We propose that phosphorylation-dephosphorylation of Rb is independent of the division cycle and is primarily determined by growth conditions throughout the division cycle. PMID- 10794603 TI - Correlation between EBV DNA and rearrangement and expression of Bcl-2 gene in aggressive non-Hodgkin's lymphoma. AB - Previous in vitro studies have shown that bcl-2 expression can be induced by transfection of Epstein-Barr virus (EBV)-negative non-Hodgkin's lymphoma (NHL) cell lines with EBV. This induced expression of bcl-2 is important for the long survival of EBV-positive cells and might be a first step in tumorigenesis. The purpose of the present study was to investigate the possibility of similar correlation between bcl-2 expression and EBV infection in vivo in a cohort of patients with aggressive NHL, who were uniformly evaluated and treated with effective chemotherapy. The 42 patients included were 25-65 years old. None had prior treatment, discordant lymphoma, or human immunodeficiency virus seropositivity. Fresh biopsied samples were obtained and stored frozen for analysis of bcl-2 gene rearrangement major break point and of EBV DNA by PCR. Bcl 2 protein expression was estimated by Western blot, and enzyme immunoassay. With a median follow-up of 30 months, overall survival (OS) and disease-free survival (DFS) were measured to determine the prognostic significance of these variables. Analyzable DNA was present in all samples, 24% demonstrating bcl-2 rearrangement and 33% showing EBV DNA. Patients with bcl-2 gene rearrangement tended to have shorter DFS, and OS than patients without translocation. Bcl-2 protein expression was not correlated to gene rearrangement and had no significant influence on survival. The presence of EBV DNA in NHL had no prognostic significance but was correlated to bcl-2 expression. EBV-positive tumors showed higher bcl-2 expression than EBV-negative tumors did. Our results suggest a role of EBV infection in inducing bcl-2 expression as a survival factor for EBV-positive cells. PMID- 10794605 TI - Surface functional properties of native, acid-treated, and reduced soy glycinin. 1. Foaming properties. AB - Foaming properties of native and chemically modified glycinin were evaluated. Effects of ionic strength and glycinin composition and concentration on foam formation and stabilization were studied. Glycinin was modified by means of combined treatments: cold or hot acidic treatments, with or without later disulfide bridges reduction. Modified proteins obtained from glycinin present different degrees of dissociation, deamidation, and as consequence, varied surface hydrophobicity and molecular size. Parameters of forming and stabilizing of foam were correlated with both deamidation and dissociation degrees of modified and native glycinin samples. A positive relationship was observed between surface behavior and foaming properties of different protein species. Results show that dissociation, deamidation, and reduction have produced structural changes on glycinin (increased surface hydrophobicity, increased net charge, decreased molecular size) which enhance the adsorption and anchorage of proteins at the air-water interface and, consequently, improve the foam forming and stabilizing capacities. PMID- 10794604 TI - Effects of vanadate on expression of liver arginase in experimental diabetic rats. AB - This work was carried out to study the effects of vanadate on the expression of liver-type arginase in experimentally induced diabetes in the rat. The results showed that the activity and mRNA levels of arginase were increased significantly in the diabetic condition. Vanadate treatment reversed the increased activity and restored mRNA levels of arginase almost to the control values. The reversal effects of vanadate were found to be similar to those of insulin, and this further confirms the insulin-like effects of vanadate. ELISA and slot-blot assay observations were consistent with the biochemical measurements of enzyme activity. The increase in arginase activity and mRNA levels in diabetes and the decrease in insulin- and vanadate-treated animals may be due to the transcriptional regulation of arginase. PMID- 10794606 TI - Surface functional properties of native, acid-treated, and reduced soy glycinin. 2. Emulsifying properties. AB - Emulsifying properties of native and chemically modified soy glycinins were studied. The influence of ionic strength, protein sample composition and concentration, and assay conditions on the flocculation-creaming process and coalescence resistance was analyzed. Differences in these emulsifying properties were exhibited by native glycinins, which have a variable content of 4S, 11S, and 15S forms. Structure and functionality of native glycinin were modified by means of combined treatments: mild acidic treatments without heating or with heating at variable time and with or without disulfide bonds reduction. Modified glycinins presented different degrees of deamidation, surface hydrophobicity, and molecular mass. A slight enhancement of emulsifying stability at moderated deamidation degrees was observed. In different protein samples, a positive relationship between the flocculation-creaming rate constant and equilibrium oil volume fraction of emulsions with surface hydrophobicity was detected. A remarkable difference was observed between reduced and nonreduced samples, mainly with respect to behavior at low or high ionic strength. PMID- 10794607 TI - Heat-induced aggregation of Phaseolus vulgaris L. Proteins: an electron spin resonance study. AB - The mechanism of heat-induced aggregation of Phaseolus vulgaris L. proteins and of subunit interactions of importance for susceptibility of proteins to proteolysis was studied by electron spin resonance (ESR) spectroscopy. The mobility of a spin label bound to lysine residues was monitored at two different pH-induced (neutral and alkaline) association states of proteins extracted from raw and cooked common bean. The molecular weight of the protein complexes was assessed by size exclusion-high performance liquid chromatography (SE-HPLC) of labeled proteins. Upon alkaline dissociation, both native and denatured protein subunits underwent a reassociation process to form soluble complexes of molecular weight higher than the species originally present at neutral pH. However, unlike native proteins, impaired mobility of the spin label was observed in the aggregates that are formed after dissociation of subunits of denatured proteins, indicating a reduced accessibility of lysine residues. Trapping of lysine residues inside protein aggregates may explain limited digestibility in the small intestine of proteins in cooked legumes. PMID- 10794608 TI - Isolation of isolectins from Vitis vinifera L. Cv. Chardonnay grape berries. AB - A lectin fraction from Chardonnay grape juice has been isolated by affinity chromatography on a column of p-aminophenyl beta-D-glucoside-derivatized agarose. The lectin fractions agglutinate rabbit and human erythrocytes without serological specificity. None of the usual monosaccharides, glycosides, or glycoproteins inhibit the hemagglutinating activity. Erythroagglutination is only inhibited by nitrophenyl glycosides, p-nitrophenyl beta-D-glucoside being the strongest inhibitor. In SDS-PAGE in the presence of 2-mercaptoethanol and gel filtration HPLC, the lectin fraction gave a single band or peak corresponding to M(r) 13.2-11.9 kDa, thus indicating it to be a monomer. Three bands were observed by isoelectric focusing with pI values of 4.1, 4. 4, and 4.9. The isolectins seem to be glycoproteins since they are bound on a concanavalin A-Sepharose column. PMID- 10794610 TI - Ultraviolet spectrophotometric evaluation of corn oil oxidative stability during microwave heating and oven test. AB - The effect of microwave heating on the oxidative stability of corn oil was determined by absorptivity in the UV spectrum and by peroxide and acid values. Oil samples with antioxidants BHA/BHT (1:1; 200 mg kg(-)(1)), with and without citric acid, were heated in a microwave oven (800 W, 2450 MHz) for 0-36 min. Absorptivity at 232 and 270 nm increased during microwave exposure. Control values of absorptivity at 232 nm increased from 3.568 to 12.874 after 36 min of heating. Peroxide value showed a significant difference in the initial stage of heating (0-6 min), but after this time, the peroxide value decreased due to the instability of hydroperoxides at high temperatures. Control 232 nm absorptivities after 6 days in the oven test were similar to those after 32-36 min of microwave heating. Effective antioxidants in the oven test did not show any protection during microwave heating. UV spectrophotometry is a suitable tool for microwave oxidation monitoring. PMID- 10794609 TI - Spanish sparkling wines (Cavas) as inhibitors of in vitro human low-density lipoprotein oxidation. AB - Forty-seven dealcoholized sparkling wines (cava) from the Penedes area in Spain were tested for their antioxidant activity in a low-density lipoprotein system. The effect of different quality-related parameters, such as harvest year or grape variety, was investigated. Twenty-two phenolic compounds were separated by high performance liquid chromatography and identified by comparing their retention time and their ultraviolet spectra with those of pure standards. When tested at the same total phenol concentration, the antioxidant activity of these white sparkling wines was found to be similar to that reported for red wines. This activity was positively correlated with the total phenolic content, trans-caffeic acid, coumaric acid, protocatechuic acid, and quercetin 3-glucuronide. The wines made of the classic cava wine coupage had superior antioxidant activity compared to those of other cultivars. PMID- 10794611 TI - Chlorophyll and carotenoid patterns in olive fruits, Olea europaea Cv. arbequina. AB - In olive fruits of the cultivar Arbequina, the chlorophyll pigments decrease significantly throughout ripening, while the carotenoids decrease more gradually and discontinuously. There is no degradation of the carotenoid fraction in stages before complete ripeness. The presence of esterified xanthophylls exclusively in this variety suggests that the chloroplast pigment metabolism is different from that in other olive varieties studied previously. There are increases of specific carotenoids, violaxanthin, neoxanthin, antheraxanthin, lutein epoxide, and esterified xanthophylls between the light green and yellowish green ripening stages. Such increases are related to the detection of precursor carotenoids (phytofluene and xi-carotene) in the yellowish green stage. Chlorophyllides (a and b) and alpha-carotene have also been detected exclusively in this variety. Quantitatively, the drastic change in color between light green and yellowish green ripening stages characteristic of this variety can be explained by the considerable reduction found in the chlorophylls/carotenoids ratio. The study of the pigments present in skin and pulp has shown that the pattern of carotenoid distribution differs depending on the fruit part concerned. PMID- 10794612 TI - Effect of postharvest storage and processing on the antioxidant constituents (flavonoids and vitamin C) of fresh-cut spinach. AB - The effect of modified atmosphere packaging (MAP) and cooking on the flavonoids and vitamin C content (ascorbic + dehydroascorbic acid; AA + DHAA) of fresh-cut spinach was evaluated. The total flavonoid content (approximately 1000 mg kg( )(1) f.w.) remained quite constant during storage in both air and MAP atmospheres, while vitamin C (750 mg kg(-)(1)f.w.) was better preserved in MAP stored spinach. AA was transformed to DHAA during storage, and its concentration was higher in MAP-stored tissues. The free-radical scavenging activity of the isolated flavonoids was tested, and only those flavonoids with either a dihydroxyl grouping or acylated with ferulic acid showed significant activity. A decrease in the total antioxidant activity was observed during storage, particularly important in MAP-stored spinach. The higher content of DHAA and lower content of both AA and antioxidant flavonoids in the MAP-stored samples could explain this antioxidant activity decrease. Boiling extracted 50% of total flavonoids and 60% vitamin C in the cooking water. However, flavonoid glucuronides were extracted more in the cooking water than the other glycosides. The vitamin C content of the cooked tissue was higher in those samples stored in MAP. PMID- 10794613 TI - Biochemical characterization of cystine lyase from broccoli (Brassica oleracea var. italica). AB - Cystine lyase is the enzyme responsible for off-aroma deterioration in fresh unblanched broccoli. In this research, cystine lyase purification from broccoli has been optimized. Only one protein peak with cystine lyase activity was found during purification. Broccoli cystine lyase was purified 100-fold to homogeneity. L-Cystine, L-cysteine-S-sulfate, L-djenkolic acid, and some S-alkyl-L-cysteines and their sulfoxides are substrates, but the enzyme had negligible activity with L-cystathionine. A K(m) value of 81.2 microM was found for L-cystine. Inhibition and K(i) determinations indicated that L-cysteine is a linear noncompetitive inhibitor with a K(i) of 5 mM and DL-homocysteine is a competitive inhibitor with a K(i) of 1.5 mM. The molecular weight of cystine lyase was determined to be 100 kDa by three methods, with two subunits of 48 kDa each and a carbohydrate content of 3%. Further characterization included cofactor quantification, the effects of temperature and pH on activity and stability, and amino acid composition. PMID- 10794614 TI - 2,2-Diphenyl-1-picrylhydrazyl radical-scavenging active components from Polygonum multiflorum thunb. AB - An activity-directed fractionation and purification process was used to identify the antioxidative components of Polygonum multiflorum Thunb. (PM). Dried root of PM was extracted with 95% ethanol and then separated into water, ethyl acetate, and hexane fractions. Among these only the ethyl acetate phase showed strong antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test when compared with water and hexane phases. The ethyl acetate fraction was then subjected to separation and purification using silica gel column chromatography and Sephadex LH-20 chromatography. Three compounds showing strong antioxidant activity were identified by spectral methods ((1)H NMR, (13)C NMR, and MS) and by comparison with authentic samples to be gallic acid, catechin, and 2,3,5, 4' tetrahydroxystilbene 2-O-beta-D-glucopyranoside. PMID- 10794615 TI - Interaction of tannin with human salivary histatins. AB - The ability of all major human salivary histatins to precipitate condensed tannin was demonstrated, and it was found that histatins 3 and 5 share the same condensed tannin-binding region but less tannin bound to histatin 1. The condensed tannin-binding region of histatin 5 includes both the N- and the C terminal parts, although more tannin binding occurs in the C-terminal region. Epigallocatechin gallate (EGCG) showed similar binding characteristics as condensed tannin, but much less EGCG was precipitated. Pentagalloyl glucose (PGG) was precipitated equally well by histatins 1, 3, and 5 and bound equally well to the N- and C-terminal regions of histatin 5. In contrast to condensed tannin, cleaving histatin 5 into N- and C-terminal fragments increased their ability to precipitate PGG. Together, these results show a number of differences in the nature of interaction of histatins with condensed tannin, EGCG, and PGG. Most of the condensed tannin-protein complexes remained insoluble under conditions similar to those in the stomach and the small intestine, suggesting that histatins may act as a defense against dietary tannin in humans. PMID- 10794617 TI - Adsorption of whey protein isolate at the oil-water interface as a function of processing conditions: a rheokinetic study. AB - In this paper we present surface dynamic properties (interfacial tension and surface dilational properties) of a whey protein isolate with a high content of beta-lactoglobulin (WPI) adsorbed on the oil-water interface as a function of adsorption time. The experiments were performed at constant temperature (20 degrees C), pH (5), and ionic strength (0.05 M). The surface rheological parameters and the interfacial tension were measured as a function of WPI concentration (ranging from 1 x 10(-)(1) to 1 x 10(-)(5)% w/w) and different processing factors (effect of convection and heat treatment). We found that the interfacial pressure, pi, and surface dilational modulus, E, increase and the phase angle, phi, decreases with time, theta, which should be associated with WPI adsorption. These phenomena have been related to diffusion of the protein toward the interface (at short adsorption time) and to the protein unfolding and/or protein-protein interactions (at long-term adsorption) as a function of protein concentration in solution and processing conditions. PMID- 10794616 TI - Nitrosation of phenolic compounds: inhibition and enhancement. AB - The nitrosation of phenol, m-, o-, and p-cresol, 2,3-, 3,5-, and 2, 6 dimethylphenol, 3,5-di-tert-butylphenol, 2,4,6-trimethylphenol, o-chlorophenol, and o-bromophenol was studied. Kinetic monitoring of the reactions was accomplished by spectrophotometric analysis of the products at 345 nm. At pH > 3, the dominant reaction was C-nitrosation through a mechanism that appears to consist of an attack on the nitrosatable substrate by NO(+)/NO(2)H(2)(+), followed by a slow proton transfer. The finding of an isokinetic relationship supports the idea that the same mechanism operates throughout the series. The observed sequence of nitrosatable substrate reactivities is explained by (i) the preferred para-orientation of the hydroxyl group for the electrophilic attack of nitrosating agents, (ii) steric hindrance of alkyl substituents, which reduces or prevents attack by nitrosating agents, and (iii) the hyperconjugative effect of the methyl substituent, which causes electronic charge to flow into the aromatic nucleus, as well as the opposite electronic withdrawing effect induced by halogen substituents. The results show that potential nitrosation of widespread environmental species such as chlorophenols is negligible, but more attention should be paid to polyphenols with strongly nucleophilic carbon atoms. PMID- 10794618 TI - Characterization of the heat treatment undergone by milk using two inhibition ELISAs for quantification of native and heat denatured alpha-lactalbumin. AB - Dairy industries are interested to know the heat treatment undergone by milk for controlling the quality of drinking milks or to control their heating systems. The purpose of this work was to develop a specific and sensitive technique for classification of the heat treatment a milk has been submitted to, without disposing of the original raw milk. For this purpose, alpha-lactalbumin was chosen as a bioindicator of heat treatment, and monoclonal antibodies specific for its native or heat-denatured form were raised and used in two inhibition ELISAs. ELISA allowed differentiation among raw, pasteurized, ultrahigh temperature-treated, and sterilized milks without even having to know the alpha lactalbumin concentration of the original raw milk. However, this technique was more suitable for intense heat treatments such as UHT treatment and sterilization because of the heat stability of alpha-lactalbumin. PMID- 10794619 TI - Effect of hexanal and iron on color development in a glucose/phenylalanine model system. AB - Aqueous glucose/phenylalanine (0.1 M with respect to each reactant) systems were heated in an autoclave for 30 min at 140 degrees C, in the presence of hexanal (0.04, 0.1, and 0.2 M) or FeCl(2) (0.01 M). Results show that hexanal significantly inhibited color development at pH 5 and 6 and led to an increase of 5-(hydroxymethyl)furfural. Iron addition had similar effects at pH 5, but only small effects at pH 6. The reaction routes proposed give key roles to the alpha dicarbonyl compounds, to the Strecker aldehyde, and to the Schiff base formed by the reaction between hexanal and phenylalanine. PMID- 10794620 TI - Molecular mechanism of the excellent emulsifying properties of phosvitin galactomannan conjugate. AB - The emulsifying properties of native and N- and C-terminal-deleted phosvitin (protease digests) were compared after conjugation with galactomannan. The emulsifying properties of Maillard-type phosvitin-galactomannan conjugates were greatly improved, whereas those of the protease-digested phosvitin-galactomannan conjugates were not so dramatically improved. Phosvitin was highly glycosylated with galactomannan, whereas the protease-digested phosvitin conjugate consisting of a highly phosphorylated core peptide fragment was not. The results suggest that both N and C termini of the peptide moiety, digested by protease, were essential for the improvement of emulsifying properties of phosvitin galactomannan conjugates. In addition, the role of N and C termini as anchors in oil droplets was supported from the comparative studies of native phosvitin, phosvitin-galactomannan conjugates, and protease-digested phosvitin-galactomannan conjugates. PMID- 10794621 TI - Lipid oxidation in emulsions as affected by charge status of antioxidants and emulsion droplets. AB - The influence of charge status of both lipid emulsion droplets and phenolic antioxidants on lipid oxidation rates was evaluated using anionic sodium dodecyl sulfate (SDS) and nonionic polyoxyethylene 10 lauryl ether (Brij)-stabilized emulsion droplets and the structurally similar phenolic antioxidants gallamide, methyl gallate, and gallic acid. In nonionic, Brij-stabilized salmon oil emulsions at pH 7.0, gallyol derivatives (5 and 500 microM) inhibited lipid oxidation with methyl gallate > gallamide > gallic acid. In the Brij-stabilized salmon oil emulsions at pH 3.0, low concentrations of the galloyl derivatives were prooxidative or ineffective while high concentrations were antioxidative. In SDS-stabilized salmon oil emulsions, oxidation rates were faster and the galloyl derivatives were less effective compared to the Brij-stabilized emulsions. Differences in antioxidant activity were related to differences in the ability of the galloyl derivatives to partition into emulsion droplets and to increase the prooxidant activity of iron at low pH. PMID- 10794622 TI - Content of the flavonols quercetin, myricetin, and kaempferol in 25 edible berries. AB - The amounts of quercetin, myricetin, and kaempferol aglycons in 25 edible berries were analyzed by an optimized RP-HPLC method with UV detection and identified with diode array and electrospray ionization mass spectrometry detection. Sixteen species of cultivated berries and nine species of wild berries were collected in Finland in 1997. Quercetin was found in all berries, the contents being highest in bog whortleberry (158 mg/kg, fresh weight), lingonberry (74 and 146 mg/kg), cranberry (83 and 121 mg/kg), chokeberry (89 mg/kg), sweet rowan (85 mg/kg), rowanberry (63 mg/kg), sea buckthorn berry (62 mg/kg), and crowberry (53 and 56 mg/kg). Amounts between 14 and 142 mg/kg of myricetin were detected in cranberry, black currant, crowberry, bog whortleberry, blueberries, and bilberry. Kaempferol was detected only in gooseberries (16 and 19 mg/kg) and strawberries (5 and 8 mg/kg). Total contents of these flavonols (100-263 mg/kg) in cranberry, bog whortleberry, lingonberry, black currant, and crowberry were higher than those in the commonly consumed fruits or vegetables, except for onion, kale, and broccoli. PMID- 10794623 TI - Volatile components in crabmeats of Charybdis feriatus. AB - The volatile components of different meats (legs with claws, body, and carapace) of a popularly consumed edible crab in Asia, Charybdis feriatus, were investigated. Samples were extracted by simultaneous steam distillation-solvent extraction and analyzed by gas chromatography/mass spectrometry. Among 177 compounds detected, 130 were positively identified. Seventy-six compounds were previously reported in other crab species. A greater number of naphthalenes were detected in this crab compared with other crabs in the literature. Aromatic compounds, alcohols, and sulfur-containing compounds were the three predominant groups with >15 components. Carapace tissue contained a greater number of volatile components in each group, except for sulfur-containing compounds. Most of the common components in the leg meat and the body meat were found at similar levels (p > 0.05). Carapace tissue generally had the highest quantity of common components among the meats. The higher levels of volatile components present in the carapace tissue might account for its stronger flavor compared with the other meats. Furthermore, the higher number of aldehydes and lower number of sulfur containing compounds detected in the carapace meat might contribute to its unique flavor. PMID- 10794624 TI - Arsenic toxicity and accumulation in turnip as affected by arsenic chemical speciation. AB - Arsenic (As) uptake by turnip, growing under soilless culture conditions, was studied. A 4 x 3 factorial experiment was conducted with four As species [arsenite, arsenate, methylarsonic acid (MMAA), dimethylarsinic acid (DMAA)] and three As concentrations (1.0, 2.0, and 5.0 mg L(-)(1)). Arsenic phytoavailability and phytotoxicity were primarily determined by As speciation. Organic arsenicals, especially MMAA, were clearly phytotoxic to this turnip cultivar. Plant As concentrations significantly increased with increasing As application rates. Both organic arsenicals showed a higher upward translocation than their inorganic counterparts, contributing to the greater phytotoxicity and lower dry matter productions of these organic treatments. Both inner root and outer root skin As concentrations were above the maximum limit set for As content in food crops (1.0 mg kg(-)(1)). If turnip plants are exposed to a large pulse of As, as growth on contaminated nutrient solutions, they will accumulate residues at levels that are unacceptable for animal and human consumption. PMID- 10794625 TI - Characterization of residual lignin after SO(2)-catalyzed steam explosion and enzymatic hydrolysis of Eucalyptus viminalis wood chips. AB - The lignin component found in both water insoluble (WI) and water and alkali insoluble (WIA) fractions derived from SO(2)-impregnated steam-exploded eucalyptus chips (SEE) was isolated and characterized. Dioxane lignins with a sugar content lower than 2% (w/w) were obtained after each material was treated with commercial cellulases. The C9 formulas of both SEE-WI and SEE-WIA dioxane lignins were C(9)H(6.83)N(0.04)O(2.24)(OCH(3))(1.21)(OH(aro))(0.56)(OH(ali))(0. 77) and C(9)H(8.65)N(0.29)O(1.97)(OCH(3))(0.90)(OH(aro))(0. 46)(OH(ali))(1.02), respectively. The weight-average molecular weight (M(w)) of the SEE-WI lignin corresponded to 3.85 kDa, whereas the SEE-WIA lignin had an M(w) of 3.66 kDa for the same polydispersity of 2.4. The SEE-WIA lignin was shown to be more thermally stable than the SEE-WI lignin, requiring temperatures in the range of 520 degrees C for complete degradation. FTIR and (1)H NMR analyses of both untreated and peracetylated lignin fractions showed that (a) the alkali insoluble lignin contained a relatively higher degree of substitution in aromatic rings per C9 unit and that (b) alkaline extraction removed lignin fragments containing appreciable amounts of phenolic hydroxyl groups. PMID- 10794626 TI - Use of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole for determining cysteine and cystine in cereal and legume seeds. AB - A new colorimetric method has been developed for the determination of chemically available cysteine and half-cystine in maize and legume seeds. The method is based on the reduction of cystine with aqueous NaBH(4)/urea/EDTA solution and the reaction of cysteine with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) in 0.2 M sodium acetate/HCl buffer (pH 2.0) to form a greenish product showing maximum absorbance at 410 nm. The method is simple, accurate, and highly specific for cysteine, in the presence of other naturally occurring amino acids. The method was applied to the determination of cysteine and of cysteine plus half-cystine in some seed meals. There was a good correlation between the results obtained using this method and those obtained using Ellman's reagent [5, 5-dithiobis(2 nitrobenzoic acid)]. There was also a good correlation between the results obtained using this method and cysteic acid values calculated from amino acid analysis of the samples. PMID- 10794627 TI - Accelerated extraction of the fat content in cheese using a focused microwave assisted soxhlet device. AB - A fast method for hydrolysis and extraction of fat in cheese based on the use of a Soxhlet extractor assisted by microwaves is proposed. The use of this type of energy dramatically reduces the time required for both steps (namely, the hydrolysis time was decreased from 1 h to 10 min with avoidance of the neutralization step, and the extraction time was decreased from 6 h to 40 min), providing similar efficiency and reproducibility comparable to or better than that of the conventional Soxhlet methodology. In addition, the proposed method is cleaner than conventional Soxhlet, as 80-85% of the extractant is recycled. The determination of the fatty acid composition by gas-liquid chromatography and quantification of both polymer formation by high-performance size exclusion liquid chromatography and nonpolar triacylglycerol formation by thin-layer chromatography with flame ionization detection in the extracts obtained by the proposed method, conventional Soxhlet and Weibull-Berntrop procedures (Soxhlet with a prior hydrolysis step), showed that the quality of the extracts obtained by the proposed procedure is better than that of the conventional alternatives, possibly due to the shorter time required by the accelerated method. PMID- 10794628 TI - Specific natural isotope profile studied by isotope ratio mass spectrometry (SNIP IRMS): (13)C/(12)C ratios of fructose, glucose, and sucrose for improved detection of sugar addition to pineapple juices and concentrates. AB - The delta(13)C values of fructose, glucose, and sucrose have been determined in authentic pineapple juices. The sugar fraction is separated from the organic acids by an anionic exchange process. Then the individual components (fructose, glucose, and sucrose) are isolated on a preparative HPLC device using a NH(2) type column. It is demonstrated that no significant isotope fractionation occurs when close to 100% of material is recovered and when the hydrolysis of sucrose is avoided. The control of the recovery rates and of the sucrose hydrolysis rate after purification is recommended for a reliable interpretation of the results. Correlations between the delta(13)C values of fructose (delta(13)Cf), glucose (delta(13)Cg), and sucrose (delta(13)Csu) can be characterized by systematic differences between these values. For the set of measurements on authentic pineapple juices and concentrates, the mean and the standard deviation of the differences are delta(13)Cf - delta(13)Cg = -0.6 +/- 0.6 per thousand, delta(13)Cf - delta(13)Csu = -1.3 +/- 0. 6 per thousand, and delta(13)Cf - delta(13)Csu = -0.7 +/- 0.5 per thousand. The determinations of the (13)C content of fructose, glucose, and sucrose enable a refinement of the detection of added sugars in fruit juices, re-enforcing the SNIP-IRMS method. PMID- 10794629 TI - Gas chromatographic organic acid profiling analysis of brandies and whiskeys for pattern recognition analysis. AB - An efficient gas chromatographic profiling and pattern recognition method is described for brandy and whiskey samples according to their organic acid contents. It involves solid-phase extraction of organic acids using Chromosorb P with subsequent conversion to stable tert-butyldimethylsilyl derivatives for the direct analysis by capillary column gas chromatography and gas chromatography mass spectrometry. A total of 12 organic acids were reproducibly identified in liquor samples (1 mL). When the GC profiles were simplified to their retention index spectra, characteristic patterns were obtained for each liquor sample as well as for each group average. Stepwise discriminant analysis provided star symbols characteristic for each liquor sample and group average. As expected, canonical discriminant analysis correctly classified 23 liquor samples studied into two groups of either brandy or whiskey. PMID- 10794630 TI - Glass transition temperature of honey as a function of water content as determined by differential scanning calorimetry. AB - The glass transition of pure and diluted honey and the glass transition of the maximally freeze-concentrated solution of honey were investigated by differential scanning calorimetry (DSC). The glass transition temperature, of the pure honey samples accepted as unadulterated varied between -42 and -51 degrees C. Dilution of honey to 90 wt % honey content resulted in a shift of the glass transition temperature by -13 to -20 degrees C. The concentration of the maximally freeze concentrated honey solutions, as expressed in terms of honey content is approximately 102-103%, i.e., slightly more concentrated in sugars than honey itself. The application of DSC measurements of and in characterization of honey may be considered, but requires systematic study on a number of honeys. PMID- 10794631 TI - Sampling leaves of young potato (Solanum tuberosum) plants for glycoalkaloid analysis. AB - Earlier attempts to measure potato (Solanum tuberosum) leaf glycoalkaloids indicated variability among similar plants, suggesting that a single small sampling of a young plant might not be a reliable measure of composition. It was also suggested that freeze-dried leaf samples might be less variable than fresh ones. In the present work, variability was minimized by comparing single leaves from the same stem position of each plant. Comparisons involving other leaves indicated that the glycoalkaloid content was not constant with respect to either time or position on the stem. In addition, the possible influence of differences in growing conditions at different times suggests that repeated plantings should include a known variety as a control to which other plants are compared. Variability was reduced by calculating glycoalkaloid concentrations on a dry weight rather than fresh weight basis. The method of drying the samples, however, had no influence on the variability of data. These considerations should be generally applicable to the sampling of replicate leaves of any plant for analysis of any components. PMID- 10794632 TI - Monitoring carbonyl-amine reaction and enolization of 1-hydroxy-2-propanone (Acetol) by FTIR spectroscopy. AB - Infrared absorption bands characteristic of the aldehydo, keto, and enediol forms of 1-hydroxy-2-propanone (acetol) were identified and used to study the effect of solvent on the absorption frequencies and the effect of temperature and acid/base catalysis on the enolization reactions. The data indicated that, in addition to water, acids and bases can catalyze the enolization of 1-hydroxy-2-propanone and that the temperature inversely effects the rate of enolization under basic conditions. However, under acidic conditions, increasing the temperature favors the enolization process. In addition, the reaction of 1-hydroxy-2-propanone with a primary and a secondary amine was also monitored by Fourier transform infrared spectroscopy. The data indicated that at room temperature the rate of amine reaction was faster than the rate of its catalysis of enolization; however, below room temperature, the rate of base-catalyzed enolization became comparable with the rate of carbonyl-amine reaction forming both Heyns and Amadori adducts. PMID- 10794633 TI - Dried green and purple lavers (Nori) contain substantial amounts of biologically active vitamin B(12) but less of dietary iodine relative to other edible seaweeds. AB - Vitamin B(12) concentrations of dried green (Enteromorpha sp.) and purple (Porphyra sp.) lavers (nori) were determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined by using the microbiological method (63.58 +/- 2.90 and 32.26 +/- 1.61 microg/100 g of dry weight) were identical to those found by using the chemiluminescence method (69.20 +/- 2.21 and 25.07 +/- 0.54 microg/100 g of dry weight) in both dried green and purple lavers, respectively. A silica gel 60 thin layer chromatography of both laver extracts shows that non-coenzyme forms (hydroxo and cyano forms) of vitamin B(12) predominate in both dried lavers. The dried lavers contained lesser amounts of dietary iodine ( approximately 4-6 mg/100 g of dry weight) relative to other seaweeds, suggesting that excessive intake of the dried lavers is unlikely to result in harmful intake of dietary iodine. These results indicate that the dried lavers (nori) are the most excellent source of vitamin B(12) among edible seaweeds, especially for strict vegetarians. PMID- 10794634 TI - Rapid purification of fumonisins and their hydrolysis products with solid-phase extraction columns. AB - Fumonisins B(3) and B(4) (FB(3) and FB(4)) were recovered from the 50:50 acetonitrile/water extract of corn cultures of a strain of Fusarium moniliforme that does not make FB(1) or FB(2) by stirring the extract with IRA-68, a weak anion-exchange resin. The fumonisins were desorbed with 5% acetic acid in the same solvent. After dilution with water, the desorbed fumonisins were separated into FB(3) (FB(3) and FA(3)) and FB(4) (FB(4), FC(4), and FA(4)) fractions with a tC(18) solid-phase extraction (SPE) cartridge. The FB(3) fraction was then separated into FB(3) and FA(3) by using an NH(2) SPE cartridge and eluting with 5% acetic acid and increasing amounts of acetonitrile in water. Finally, FB(1) and FA(3) were hydrolyzed with calcium hydroxide. After recovery from the reaction mixture using a tC(18) cartridge, the hydrolyzed and partially hydrolyzed analogues were separated and the unreacted fumonisins recovered by using an NH(2) cartridge, initially in the normal-phase mode with increasing amounts of water in acetonitrile and then in the reversed-phase mode after the addition of 5% acetic acid to the solvent and eluting in the reverse order. PMID- 10794636 TI - Volatile compounds from potato-like model systems. AB - The volatile reaction products of aqueous mixtures comprising combinations of methionine, glucose, linoleic acid, and starch heated in a modified Likens Nickerson apparatus were extracted and analyzed by gas chromatography-mass spectrometry (GC-MS). The majority of volatile compounds were formed from linoleic acid degradation, hexanal, 2,4-decadienal, and 2-pentylfuran being identified in the greatest amounts. Dimethyl disulfide and dimethyl trisulfide were detected in every system containing methionine. 3-(Methylthio)propanal (methional) and other sulfur compounds were detected when methionine was heated with another precursor. No binding of volatile compounds to starch was observed; rather, starch appeared to act as an additional source of reactive carbohydrate. Almost all the components identified have been identified among the aroma components of cooked potato. No pyrazines, pyridines, or thiazoles were identified, probably due to the relatively low temperature/high moisture conditions. PMID- 10794635 TI - Effects of tea polyphenols on the invasion and matrix metalloproteinases activities of human fibrosarcoma HT1080 cells. AB - The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity. PMID- 10794637 TI - Production of seafood flavor from red hake (Urophycis chuss) by enzymatic hydrolysis. AB - Protein hydrolysates were prepared as a natural flavor stock from the red hake (Urophycis chuss) headed-gutted (H&G) mince and frame mince using commercial enzymes, Flavourzyme and Savorase, at the natural pH of fish (6.8) and the water/fish ratio of 2:5. The addition of 1.5% NaCl and 0.4% STPP improved the flavor quality of the hydrolysate by masking bitterness and off-flavor. A 6 h hydrolysis of H&G mince with Flavourzyme yielded a hydrolysate of the highest acceptability. Hydrolysis increased the concentration of most free amino acids except Arg and His. Leu, Lys, and Arg were predominant free amino acids in the hydrolysates, whereas Leu and Arg were major ones in the cooking juice. The concentration of Glu responsible for umami taste was increased by 6-9 times upon hydrolysis. Hydrolysates contained higher percentages of free amino acids giving both umami and sweet tastes than did cooking juice. PMID- 10794638 TI - Distribution of bromophenols in species of marine algae from eastern Australia. AB - Forty-nine species (87 samples) of marine macroalgae from eastern Australia were analyzed by GC/MS for the key seafood flavor components 2- and 4-bromophenol, 2,4 and 2,6-dibromophenol, and 2,4, 6-tribromophenol. All five bromophenols were found in 62% of samples, four in 32% of samples, and three in the remaining 6% of samples. 2, 4,6-Tribromophenol was found in all samples and, with few exceptions, was present in the highest concentrations. The total bromophenol content determined on a wet-weight basis varied widely across species from 0.9 ng/g in the green alga Codium fragile to 2590 ng/g in the red alga Pterocladiella capillacea. Species with the highest concentrations of bromophenols were all collected from sites exposed at low tide. The study demonstrates the wide occurrence of bromophenols in marine algae and provides a possible source of such compounds in fish that feed predominantly on ocean plants. The possible effect that dietary marine algae has on the flavor of omnivorous ocean fish is discussed. PMID- 10794639 TI - Contribution of 3-methylthiopropionaldehyde to the worty flavor of alcohol-free beers. AB - Alcohol-free beers are usually criticized for two major defects: a lack of fruity aroma and a strong worty flavor. 3-Methylbutanal and 2-methylbutanal are described in the literature as being predominantly responsible for the worty taste. Although detected in large amounts in most malt and wort extracts, both compounds have proven unable to confer worty taste to beers. In this work, we extracted volatiles from wort with a Likens-Nickerson microextractor. The resulting extract had a strong worty aroma. Following GC/MS and GC/olfactometry analysis, 3-methylthiopropionaldehyde turned out to be the key feature since it remained the most organoleptically active compound through extract dilution. In vitro assays showed that 3-methylthiopropionaldehyde is a substrate for several Saccharomyces cerevisiae reducing enzymes. Screening of various strains led us to conclude that the higher the 3-methylbutanal reductase activity, the higher the 3 methylthiopropionaldehyde reductase activity. Monitoring of 3 methylthiopropionaldehyde concentration through an in vivo cold contact process has also been undertaken. PMID- 10794640 TI - Aroma comparisons of traditional and mild yogurts: headspace gas chromatography quantification of volatiles and origin of alpha-diketones. AB - A quick headspace GC method for quantification of volatiles was developed, involving only minor sample preparation. Yogurt flavor compounds could be quantified in the micrograms per kilogram to milligrams per kilogram range without any difficulty, despite the complex matrix. Volatiles of traditional acidic and mild, less acidic yogurts were compared, and important differences were found for acetaldehyde, 2,3-butanedione, and 2,3-pentanedione. Concentrations of 2,3-butanedione and 2,3-pentanedione increased 2-3-fold in mild, less acidic yogurts compared to traditional acidic ones. This is due to accumulation of the precursors of the diketones, 2-acetolactate and 2 acetohydroxybutyrate, during fermentation in mild, less acidic yogurt. These precursors are subsequently converted to the corresponding diketones during storage. On the contrary, acetaldehyde formation was reduced in the mild yogurt, due to growth differences between the lactic acid bacteria used for fermentation of the milk. The quantitative results presented in this study validate previous GC sniffing conclusions (Ott et al. J. Agric. Food Chem. 1997, 45, 850-858), showing that yogurt aroma is the superposition of impact flavor compounds generated by fermentation on milk compounds. PMID- 10794641 TI - Effect of various cryostabilizers on the production and reactivity of formaldehyde in frozen-stored minced blue whiting muscle. AB - The production of formaldehyde in frozen-stored minced blue whiting muscle was described by a rectangular hyperbolic model, and the effectiveness of each cryostabilizer is discussed in terms of its parameters. The maltodextrins assayed noticeably inhibited formaldehyde production, this effect being greater at -20 degrees C than at -10 degrees C. Sucrose was only effective at -20 degrees C. It seems that these compounds act by restricting molecular diffusion. The effect of each cryostabilizer on formaldehyde binding was closely regulated by its effect on production. This is discussed in terms of the binding equation parameters. The binding of formaldehyde during frozen storage was dependent on protein rearrangements leading to reactive groups becoming available. The constraints of cryostabilizers on molecular diffusion reduced the exposure of these groups. Consequently, the interpretation of formaldehyde reactivity was biased, leading to conclusions different from those that would be obtained from a study done under standard conditions. PMID- 10794642 TI - Phenolic composition of kiwifruit juice. AB - Phenolic compounds in kiwifruit pulp were separated and characterized by reversed phase HPLC, and the effect of juice processing on the phenolic composition was studied. Fractionation of phenolic compounds was achieved through selective elution from C-18 cartridges prior to preconcentration and subsequent separation by HPLC. Strongly acidic compounds were identified as derivatives of coumaric and caffeic acids, including chlorogenic acid, protocatechuic acid, and a derivative of 3,4-dihydroxybenzoic acid. The weakly acidic fraction contained epicatechin, catechin, and procyanidins (B3, B2, or B4 and oligomers). Flavonols were present as the glycosides of quercetin (glucoside, rhamnoside, and rutinoside) and kaempferol (rhamnoside and rutinoside). Phenolic compounds were present, at levels of <1-7 mg/L, in clarified juice. The concentration of phenolics was highest after high-temperature short-time treatment (HTST) of juice. Hydrolysis of hydroxycinnamic acids occurred after enzyme addition and HTST treatment. The flavonol glycoside composition is the best identifier of kiwifruit juice. PMID- 10794643 TI - Kinetics of chlorophyll degradation and color loss in heated broccoli juice. AB - Degradation of chlorophyll in broccoli juice occurred at temperatures exceeding 60 degrees C. Chemical analysis revealed that degradation of chlorophyll a and b to pheophytin a and b, respectively, followed first-order kinetics and that chlorophyll a was more heat sensitive than chlorophyll b. Temperature dependencies of chlorophyll a and b degradation rate constants could be described by Arrhenius equations with activation energies (E(a)) of 71.04 +/- 4.89 and 67.11 +/- 6.82 kJ/mol, respectively. Objective greenness measurements, using the a value as the physical property, together with a fractional conversion kinetic analysis, indicated that green color degradation followed a two-step process. Kinetic parameters for the first degradation step were in accordance with the kinetic parameters for pheophytinization of the total chlorophyll content, as determined by chemical analysis (E(a) approximately 69 kJ/mol). The second degradation step, that is, the subsequent decomposition of pheophytins, was characterized by an activation energy of 105.49 +/- 4.74 kJ/mol. PMID- 10794644 TI - Catalase in the heat-induced chilling tolerance of cold-stored hybrid Fortune mandarin fruits. AB - Hybrid Fortune mandarins developed chilling injury (CI) upon cold storage, unless the fruits were conditioned at 37 degrees C for 3 days before they were held at low temperature. This heat treatment induced 2.5-, 1.2-, and 1.4-fold increases in the activities of catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD), respectively, and reduced the activity of glutathione reductase (GR). The differences in the activities afforded by the heat treatment were, in general, maintained during cold storage. However, SOD levels in nonconditioned Fortune fruits exhibiting CI were similar to those of conditioned fruits stored for 0 or 6 weeks at 2 degrees C. No difference between APX activity in the conditioned and nonconditioned fruits stored for 6 weeks at 2 degrees C was found. The data indicate that CAT may be a major antioxidant enzyme operating in the heat-induced chilling tolerance of cold-stored Fortune mandarin fruits. PMID- 10794645 TI - Sensory attribute variation in low-temperature-stored roasted peanut paste. AB - Length of sample storage can become significant in sensory studies due to panel fatigue limitations and samples needed for a reasonable expectation of finding significant differences. In roasted peanut sensory studies samples are stored between -10 and -23 degrees C to prevent or retard changes. Studies of up to 13 months' duration have examined stability and slow-rate sensory changes. Sweet taste was relatively stable, whereas bitter and tongue burn attributes increased slightly. Stale taste increased, suggesting lipid oxidation was taking place even at -23 degrees C. Painty attribute did not increase until stale was >3. An increase in fruity attribute was unexpected. With increases in fruity and stale attributes a decrease in roasted peanut was expected. However, storage at -23 degrees C seems to stabilize the roasted peanut lability when compared to storage at -10 degrees C. Fruity and stale interactions with roasted peanut and lability of roasted peanut were shown to be three separate and identifiable effects on roasted peanut. PMID- 10794646 TI - Fate of xanthohumol and related prenylflavonoids from hops to beer. AB - The fate of three prenylated flavonoids of the chalcone type, xanthohumol, desmethylxanthohumol, and 3'-geranylchalconaringenin, was monitored with LC/MS-MS from hops (Humulus lupulus L.) to beer in two brewing trials. The three prenylchalcones were largely converted into their isomeric flavanones, isoxanthohumol, prenylnaringenins, and geranylnaringenins, respectively, in the boiling wort. Losses of prenylflavonoids were due to incomplete extraction from the hops into the wort (13-25%), adsorption to insoluble malt proteins (18-26%), and adsorption to yeast cells (11-32%) during fermentation. The overall yield of xanthohumol, after lagering of the beer and largely in the form of isoxanthohumol, amounted to 22-30% of the hops' xanthohumol. About 10% of the hops' desmethylxanthohumol, completely converted into prenylnaringenins, remained in the beers. 3'-Geranylchalconaringenin behaved similarly to desmethylxanthohumol. Solubility experiments indicated that (1) malt carbohydrates form soluble complexes with xanthohumol and isoxanthohumol and (2) solubility does not dictate the isoxanthohumol levels of finished beers. PMID- 10794647 TI - Metabolism of 7-fluoro-6-(3,4,5,6-tetrahydrophthalimido)-4- (2-propynyl)-2H-1,4 benzoxazin-3(4H)-one (S-53482, flumioxazin) in the rat: II. Identification of reduced metabolites. AB - On single oral administration of (14)C-S-53482 [7-fluoro-6-(3,4,5, 6 tetrahydrophthalimido)-4-(2-propynyl)-2H-1,4-benzoxazin-3( 4H)-one, Flumioxazin] labeled at the 1- and 2-positions of tetrahydrophthaloyl group to rats at 1 (low dose) or 100 (high dose) mg/kg, the radiocarbon was almost completely eliminated within 7 days after administration in both groups with generally very low residual (14)C tissue levels. The predominant excretion route was via the feces. The major fecal and urinary metabolites involved reduction or sulfonic acid addition reactions at the 1,2-double bond of the 3,4,5,6-tetrahydrophthalimide moiety and hydroxylation of the cyclohexene or cyclohexane ring. One urinary and four fecal metabolites were identified using chromatographic techniques and spectroanalyses (NMR and MS). Three of five identified metabolites were unique forms, reduced at the 1,2-double bond of the 3,4,5, 6-tetrahydrophthalimide moiety. On the basis of the metabolites identified in this study, the metabolic pathways of S-53482 in rats are proposed. To specify tissues forming reduced metabolites, an in vitro study was conducted. Reduction was found to take place in red blood cells. PMID- 10794648 TI - Metabolism of [(14)C]flusilazole in the goat. AB - [Phenyl(U)-(14)C] and [triazole(3)-(14)C]flusilazole ([(bis 4 fluorophenyl)]methyl(1H-1,2,4-triazole-1-ylmethyl)silane; I) were extensively metabolized when fed to lactating goats (Capra hircus). The primary metabolites identified in goat tissues and milk were bis(4-fluorophenyl)(methyl)silanol (II) and 1H-1,2,4-triazole (III). Concentrations of total radiolabeled residues in the milk ranged from 0.09 to 0.74 microg/mL. Concentrations of radiolabeled residues found in tissues when the [(14)C] label was in the phenyl or triazole position, respectively, were 13.5 and 3.54 microg/g (liver), 8.74 and 0.75 microg/g (kidney), 0.41 and 0.52 microg/g (leg muscle), and 4.07 and 0.94 microg/g (back fat). Urine contained an additional major metabolite identified as [bis(4 fluorophenyl)](methyl)silylmethanol (IV) and its glucuronic acid conjugate (V). With either labeled form of flusilazole, the majority of the recovered radiolabel was excreted in urine or feces. PMID- 10794649 TI - Volatiles from Fusarium verticillioides (Sacc.) nirenb. and their attractiveness to nitidulid beetles. AB - It is known that sap beetles (Coleoptera: Nitidulidae) can vector the fungus Fusarium verticillioides (Sacc.) Nirenb. (= F. moniliforme Sheldon), which causes an important ear-rot disease in corn and also produces fumonisin mycotoxins. The volatiles produced by this fungus were studied to establish whether they could attract sap beetles. Such an association would suggest more than just an incidental role in transmission of the fungus by the beetles. F. verticillioides consistently produces a blend of five alcohols (ethanol, 1-propanol, 2-methyl-1 propanol, 3-methyl-1-butanol, and 2-methyl-1-butanol), acetaldehyde, and ethyl acetate. Ethanol is the most abundant alcohol. The fungus also produces four phenolic compounds (the most abundant of which is ethylguaiacol), a series of presently unidentified sesquiterpene hydrocarbons, and an unidentified compound that is probably a 10-carbon ketone. Solid-phase microextraction was the key technique used in volatile analysis. The volatile profiles change over time and differ somewhat among fungal strains: The alcohols, aldehyde, and ester always appeared first and were present for each strain. Production of the phenolics lagged by several days, and in some strains these compounds were barely detectable. Volatile production eventually diminished in all strains. All strains were attractive to the sap beetle, Carpophilus humeralis (F.), in wind-tunnel bioassays. Attraction was correlated primarily to the presence of the alcohols, acetaldehyde, and ethyl acetate, rather than to the phenolics. To verify that the identified culture volatiles were responsible for beetle attraction, cultures were quantitatively simulated with synthetic chemicals, and the cultures and corresponding synthetic mixtures were then compared by bioassay. The comparisons were favorable. Volatile emission patterns from cultures were fairly robust with respect to inoculum level or incubation temperature, but some manipulation was possible. For example, after freeze-drying and rehydrating (a rapid simulation of winter/spring conditions), F. verticillioides produced ethyl acetate and other esters at unusually high levels. The fungus produced attractive volatiles following ear inoculation of milk-stage field corn as well as on sterile, mature kernels in the laboratory. PMID- 10794650 TI - Potential environmental impact of effluents from the artichoke (Cynara scolymus L.) byproduct ensiling process using additives. AB - Three treatments have been tested on canned artichoke byproduct after 50 days of ensilage: formic acid at 20% in doses of 2 mL. kg(-)(1) (FA), cane sugar molasses at 50 g.kg(-)(1) (M), and sodium chloride at 30 g.kg(-)(1) (SC). A fourth batch acted as a control group (C). The nutritive value, fermentation characteristics, environmental pollution effect, and total volume of effluents released have been studied. The highest nutritive value recorded was with SC silage. The use of the additives did not significantly improve the fermentation stability of the silage, but the total production of effluents in each treatment-52.7 (FA), 46.9 (M), and 55.2 (SC)-was significantly lower (P < 0.01) than that of the control group (70.1 L.Tm(-)(1)). The chemical oxygen demand (COD), 117300 mg of O(2).L(-)(1), and the conductivity, 46.4 microOmega(-)(1). cm(-)(1), were significantly higher (P < 0.01) in M and SC, respectively, than in the other group. PMID- 10794651 TI - Influence of incubation temperature on the behavior of triethylamine-extractable glyphosate (N-phosphonomethylglycine) in four soils. AB - The behavior of glyphosate, extracted from four soils using aqueous triethylamine, was investigated at two temperatures. For each soil, and at both temperatures, there was a marked loss in the amount of extractable glyphosate immediately after addition of the herbicide to soil. This rapid loss of glyphosate was ascribed to adsorption of the herbicide into a nonextractable form. For three of the four soils used when incubated at 25 degrees C, the rates of loss of extractable glyphosate were similar to previously measured rates of degradation of this herbicide in these soils. However, loss of extractable glyphosate from the Culgoa clay loam was due not only to substrate degradation but also to slow sorption of glyphosate into the nonextractable form in this soil over the experimental period. For the Rutherglen and Walpeup soils, when incubated at 10 degrees C, the rates of loss of extractable glyphosate were half of the previously measured rate of degradation of this herbicide in these soils. However, there was no measured loss of extractable glyphosate from the Wimmera clay. As previous work has shown glyphosate to decompose readily in these soils at this temperature, these findings suggest that desorption of glyphosate may occur at a rate greater than degradation at this temperature and, hence, that temperature may play a pivotal role in sorption processes. Investigations with these soils when sterilized by gamma-irradiation showed that for the Walpeup, Wimmera, and Rutherglen soils, sorption was complete soon after the addition of the herbicide; however, for the Culgoa soil, further adsorption occurred over the entire experimental period. PMID- 10794652 TI - Soybean lipoxygenase inactivation by pressure at subzero and elevated temperatures. AB - Soybean lipoxygenase (LOX) inactivation [0.4 mg/mL in Tris-HCl buffer (0.01 M, pH 9)] was studied quantitatively under constant pressure (up to 650 MPa) and temperature (-15 to 68 degrees C) conditions and kinetically characterized by rate constants, activation energies, and activation volumes. The irreversible LOX inactivation followed a first-order reaction at all pressure-temperature combinations tested. In the entire pressure-temperature area studied, LOX inactivation rate constants increased with increasing pressure at constant temperature. On the contrary, at constant pressure, the inactivation rate constants showed a minimum around 30 degrees C and could be increased by either a temperature increase or decrease. On the basis of the calculated rate constants at 102 pressure-temperature combinations, an iso-rate contour diagram was constructed as a function of pressure and temperature. The pressure-temperature dependence of the LOX inactivation rate constants was described successfully using a modified kinetic model of Hawley. PMID- 10794653 TI - Development of monoclonal antibody-based immunoassays to the N-methylcarbamate pesticide carbofuran. AB - To produce monoclonal antibodies (MAbs) to the pesticide carbofuran, three compounds with carboxylic spacer arms of different lengths introduced at the carbamate group of the analyte structure were synthesized, conjugated to proteins, and used as immunizing haptens in mice. MAbs were subsequently characterized for affinity and specificity in the conjugate-coated format and in the antibody-coated format using newly synthesized compounds as heterologous assay haptens. Depending on the immunoreagent combination and assay format, competitive assays with I(50) values in the 1.2-10.2 nM (0.27-2.27 ng/mL) range were obtained. LIB-BFNB67 MAb in combination with the hapten BFNH, coupled either to horseradish peroxidase or to ovalbumin, was used to develop a direct and an indirect enzyme-linked immunosorbent assay, respectively. Optimized immunoassays displayed very similar analytical characteristics, with an I(50) value around 0.7 ng/mL and a limit of detection around 0.08 ng/mL. Both immunoassays were able to tolerate the presence of methanol up to a 15% concentration. Compounds very similar in structure to carbofuran (benfuracarb, furathiocarb, bendiocarb, and carbofuran-hydroxy) exhibited cross-reactivity values in the 18-37% range, but major N-methylcarbamate pesticides were not recognized by the MAb. These immunoassays should reasonably allow the rapid, low-cost, and sensitive determination of carbofuran in food, in soils, and in the environment at levels of regulatory and practical importance. PMID- 10794654 TI - Characterization of tyrosinase- and polyphenol esterase-catalyzed end products using selected phenolic substrates. AB - The oxidative end products that result from the biocatalysis of tyrosinase (PPO) and/or a polyphenol esterase (PPE) extract have been investigated simultaneously in model systems containing selected phenolic compounds as substrates. The spectrophotometric scanning of brown color, formed in the presence of both PPO and PPE, showed a decrease in the absorbance compared to that obtained with PPO only. Graphical analyses of the iterative spectra of oxidized phenolic end products by PPO confirmed the presence of, at least, three kinetically related absorbing species. HPLC analyses of the end products, obtained by the biocatalysis of PPE or PPO activity, indicated the presence of two main groups of compounds: colored ones of lambda(max) at 294-324 nm and colorless products of lambda(max) at 264-290 nm. PPE produced both compounds when selected substrates were used as substrates, whereas PPO produced only one type of oxidation product. However, when both enzymes were incubated together, the nature of the end products was similar to that obtained with PPE only. PMID- 10794655 TI - Production of organically bound selenium yeast by continuous fermentation. AB - This paper describes a protocol for incorporation of sodium selenite or sodium selenate into Saccharomyces cerevisiae biomass by continuous fermentation in a medium with minimal sulfur and methionine concentrations. Selenium incorporation was followed by atomic absorption analysis and methylene blue reduction time (MBRT). Continuous fermentation at 0.2 h(-1) dilution rate and sodium selenite addition gradient up to 0.69 g/L of Na(2)SeO(3) yielded 1. 89 g/L of biomass with 1904 microg of selenium/g of dry biomass. However, MBRT was 0.1 min, which indicated that the majority of selenium was in the inorganic form. On the other hand, continuous fermentation at 0.2 h(-1) dilution rate and sodium selenate gradient up to 0.28 g/L of Na(2)SeO(4) yielded 0.76 g/L of dry biomass with 687 microg of selenium/g of dry biomass, and MBRT was 26 min, which indicated a high concentration of organically bound selenium. Overall, the results indicate a Se/S ratio of 3.9:1 and a dry biomass/Se ratio of 5.5:1 as optimal for continuous production of organically bound selenium. PMID- 10794656 TI - Enhanced organically bound selenium yeast production by fed-batch fermentation. AB - This study describes a fed-batch fermentation protocol for enhanced production of Saccharomyces cerevisiae containing organically bound selenium. Two levels of sodium selenate concentrations were applied as either a single dose or continuous addition. Fermentations with high sodium selenate (63.2 g/L in cane molasses feeding medium) demonstrated 24 g/L of biomass with 1382 microg of selenium/g of dry biomass for single-dose addition and 40 g/L of biomass with 1491 microg of selenium/g of dry biomass for continuous addition. Low selenium concentration (31.6 g/L in cane molasses feeding medium) demonstrated higher biomass concentration with higher selenium level; 37 g/L of biomass with 2846 microg of selenium/g of dry biomass and 45 g/L of biomass with 2495 microg of selenium/g of dry biomass for single-dose and continuous addition, respectively. Also, two adapted S. cerevisiae strains were evaluated in fed-batch fermentation. A single dose of low concentration demonstrated >3000 microg of selenium/g of dry biomass, but biomass concentration was lower (< or =32 g/L) for these adapted strains. PMID- 10794658 TI - KChAP: a novel chaperone for specific K(+) channels key to repolarization of the cardiac action potential. Focus on "KChAP as a chaperone for specific K(+) channels". PMID- 10794659 TI - Inhibition of phosphatidylinositol 3-kinase does not alter forskolin-stimulated Cl(-) secretion by T84 cells. AB - Wortmannin is a potent inhibitor of phosphatidylinositol 3-kinase (PI3K) and membrane trafficking in many cells. To test the hypothesis that cystic fibrosis transmembrane conductance regulator (CFTR) traffics into and out of the plasma membrane during cAMP-stimulated epithelial Cl(-) secretion, we have studied the effects of wortmannin on forskolin-stimulated Cl(-) secretion by the human colonic cell line T84. At the PI3K inhibitory concentration of 100 nM, wortmannin did not affect significantly forskolin-stimulated Cl(-) secretion measured as short-circuit current (I(SC)). However, 500 nM wortmannin significantly inhibited forskolin-stimulated I(SC). cAMP activation of apical membrane CFTR Cl(-) channels in alpha-toxin-permeabilized monolayers was not reduced by 500 nM wortmannin, suggesting that inhibition of other transporters accounts for the observed reduction in T84 Cl(-) secretion. Forskolin inhibits apical endocytosis of horseradish peroxidase (HRP), but wortmannin did not alter forskolin inhibition of apical HRP endocytosis. In the absence of forskolin, wortmannin stimulated HRP endocytosis significantly. We conclude that, in T84 cells, apical fluid phase endocytosis is not dependent on PI3K activity and that CFTR does not recycle through a PI3K-dependent and wortmannin-sensitive membrane compartment. PMID- 10794660 TI - Chlorpromazine modulates the morphological macro- and microstructure of endothelial cells. AB - Chlorpromazine (CP), an amphipathic, antipsychotic agent, causes concave membrane bending in red blood cells with formation of stomatocytic shapes by modulation of the phospholipid bilayer. This study was designed to investigate the effects of CP on the shape of bovine aortic endothelial cells (BAEC) and their membranes in confluent monolayers with phase-contrast and transmission electron microscopy. Exposure of BAECs to nanomolar levels of CP leads to membrane curvature changes. With increasing CP concentrations, the membrane assumed a shape with enhanced numbers of intracellular caveolae and projection of pseudopodia at all junctions. At higher CP concentrations (up to 150 microM), the endothelial cells assumed almost spherical shapes. The evidence suggests that CP may affect lipid bilayer bending of BAECs in analogy with previous observations on erythrocytes, supporting the formation of caveolae and pseudopodia in BAECs due to the induction of concave membrane bending, as well as an effect on endothelial cell membrane adhesion at higher CP concentrations with loss of cellular attachment at junctions. PMID- 10794661 TI - Ca(2+) and p38 MAP kinase regulate mAChR-mediated c-Fos expression in avian exocrine cells. AB - Muscarinic acetylcholine receptors (mAChRs) in exocrine tissue from the avian nasal salt gland are coupled to phospholipase C and generate inositol phosphate and Ca(2+) signals upon activation. An early effect of receptor activation in the secretory cells is a transient accumulation of c-Fos protein. In cooperation with constitutively expressed Jun, Fos presumably serves as a transcription factor altering gene expression during cell growth and differentiation processes in the gland associated with adaptation to osmotic stress in animals. Nothing is known, however, about the mAChR-dependent signaling pathways leading to Fos expression in these cells. By incubation of isolated nasal gland tissue in short-term culture with activators or inhibitors of signaling pathways and quantitative Western blot analysis of Fos abundance, we have now identified the sustained elevation of the intracellular Ca(2+) concentration and the activation of the p38 mitogen-activated protein (MAP) kinase as intermediate signaling elements for the regulation of c-Fos by muscarinic receptor activation. It is suggested that p38 MAP kinase, rather than exclusively mediating stress responses, is involved in the regulation of cellular growth and differentiation controlled by G protein coupled receptors. PMID- 10794662 TI - Ovine male genital duct epithelial cells differentiate in vitro and express functional CFTR and ENaC. AB - To investigate the biology of the male genital duct epithelium, we have established cell cultures from the ovine vas deferens and epididymis epithelium. These cells develop tight junctions, high transepithelial electrical resistance, and a lumen-negative transepithelial potential difference as a sign of active transepithelial ion transport. In epididymis cultures the equivalent short circuit current (I(sc)) averaged 20.8+/-0.7 microA/cm(2) (n = 150) and was partially inhibited by apical application of amiloride with an inhibitor concentration of 0.64 microM. In vas deferens cultures, I(sc) averaged 14.4+/-1.1 microA/cm(2) (n = 18) and was also inhibited by apical application of amiloride with a half-maximal inhibitor concentration (K(i)) of 0.68 microM. The remaining amiloride-insensitive I(sc) component in epididymis and vas deferens cells was partially inhibited by apical application of the Cl(-) channel blocker diphenylamine-2-carboxylic acid (1 mM). It was largely dependent on extracellular Cl(-) and, to a lesser extent, on extracellular HCO(-)(3). It was further stimulated by basolateral application of forskolin (10(-5) M), which increased I(sc) by 3.1+/-0.3 microA/cm(2) (n = 65) in epididymis and 0.9+/-0.1 microA/cm(2) (n = 11) in vas deferens. These findings suggest that cultured ovine vas deferens and epididymis cells absorb Na(+) via amiloride-sensitive epithelial Na(+) channels (ENaC) and secrete Cl(-) and HCO(-)(3) via apical cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels. This interpretation is supported by RT-PCR data showing that vas deferens and epididymis cells express CFTR and ENaC mRNA. PMID- 10794663 TI - Mechanical strain memory in airway smooth muscle. AB - We investigated the effect of a single rapid stretch on poststretch force and myosin phosphorylation in bovine tracheal smooth muscle. When unstimulated muscle strips were stretched from suboptimal length to optimal length (L(o)), poststretch steady-state force was not significantly different from that of unstretched control at L(o). However, when carbachol-activated muscle strips were stretched from suboptimal length to L(o), poststretch force and myosin phosphorylation were lower than control and significantly correlated with initial length. When poststretch muscle strips were allowed to relax for 1 h and then activated by K(+) depolarization, the developed force remained significantly correlated with initial length. When the same strain was applied in 23 increments to minimize peak stress, poststretch force and myosin phosphorylation increased significantly, approaching the levels expected at L(o). Furthermore, poststretch force development increased after each cycle of contraction and relaxation, approaching the control level after four cycles. These results suggest that activated airway smooth muscle cells can retain relatively precise memory of past strain when they are stretched rapidly with high stress. PMID- 10794664 TI - Vasopressin and PGE(2) regulate activity of apical 70 pS K(+) channel in thick ascending limb of rat kidney. AB - Vasopressin and prostaglandin E(2) (PGE(2)) are involved in regulating NaCl reabsorption in the thick ascending limb (TAL) of the rat kidney. In the present study, we used the patch-clamp technique to study the effects of vasopressin and PGE(2) on the apical 70 pS K(+) channel in the rat TAL. Addition of vasopressin increased the channel activity, defined as NP(o), from 1.11 to 1.52 (200 pM) and 1.80 (500 pM), respectively. The effect of vasopressin can be mimicked by either forskolin (1-5 microM) or 8-bromo-cAMP/dibutyryl-cAMP (8-Br-cAMP/DBcAMP) (200-500 microM). Moreover, the effects of cAMP and vasopressin were not additive and application of 10 microM H-89 abolished the effect of vasopressin. This suggests that the effect of vasopressin is mediated by a cAMP-dependent pathway. Applying 10 nM PGE(2) alone had no significant effect on the channel activity. However, PGE(2) (10 nM) abolished the stimulatory effect of vasopressin. The PGE(2) induced inhibition of the vasopressin effect was the result of decreasing cAMP production because addition of 200 microM 8-Br-cAMP/DBcAMP reversed the PGE(2) induced inhibition. In addition to antagonizing the vasopressin effect, high concentrations of PGE(2) reduced channel activity in the absence of vasopressin by 33% (500 nM) and 51% (1 microM), respectively. The inhibitory effect of high concentrations of PGE(2) was not the result of decreasing cAMP production because adding the membrane-permeant cAMP analog failed to restore the channel activity. In contrast, inhibiting protein kinase C (PKC) with calphostin C (100 nM) abolished the effect of 1 microM PGE(2). We conclude that PGE(2) inhibits apical K(+) channels by two mechanisms: 1) low concentrations of PGE(2) attenuate the vasopressin-induced stimulation mainly by reducing cAMP generation, and 2) high concentrations of PGE(2) inhibit the channel activity by a PKC-dependent pathway. PMID- 10794665 TI - Localization of two distinct type III phosphatidylinositol 4-kinase enzyme mRNAs in the rat. AB - Inositol lipid kinases generate polyphosphoinositides, important regulators of several cellular functions. We have recently cloned two distinct phosphatidylinositol (PI) 4-kinase enzymes, the 210-kDa PI4KIIIalpha and the 110 kDa PI4KIIIbeta, from bovine tissues. In the present study, the distribution of mRNAs encoding these two enzymes was analyzed by in situ hybridization histochemistry in the rat. PI4KIIIalpha was found predominantly expressed in the brain, with low expression in peripheral tissues. PI4KIIIbeta was more uniformly expressed being also present in various peripheral tissues. Within the brain, PI4KIIIbeta showed highest expression in the gray matter, especially in neurons of the olfactory bulb and the hippocampus, but also gave a signal in the white matter indicating its presence in glia. PI4KIIIalpha was highly expressed in neurons, but lacked a signal in the white matter and the choroid plexus. Both enzymes showed expression in the pigment layer and nuclear layers as well as in the ganglion cells of the retina. In a 17-day-old rat fetus, PI4KIIIbeta was found to be more widely distributed and PI4KIIIalpha was primarily expressed in neurons. These results indicate that PI4KIIIbeta is more widely expressed than PI4KIIIalpha, and that the two enzymes are probably coexpressed in many neurons. Such expression pattern and the conservation of these two proteins during evolution suggest their nonredundant functions in mammalian cells. PMID- 10794666 TI - Monocarboxylic acid transporters, MCT1 and MCT2, in cortical astrocytes in vitro and in vivo. AB - We used sequence-specific antibodies to characterize two monocarboxylic acid transporters, MCT1 and MCT2, in astrocytes. Both proteins are expressed in primary cultures of cortical astrocytes, as indicated by immunoblotting and immunofluorescence. Both MCT1 and MCT2 are present in small, punctate structures in the cytoplasm and at the cell membrane. Cells showing very low levels of labeling for glial fibrillary acidic protein (GFAP) also label more dimly for MCT2, but not for MCT1. In vivo, double-label immunofluorescence studies coupled with confocal microscopy indicate that MCT1 and MCT2 are rare in astrocytes in the cortex. However, they are specifically labeled in astrocytes of the glial limiting membrane and in white matter tracts. Both transporters are also present in the microvasculature. Comparison of labeling for MCT1 and MCT2 with markers of the blood-brain barrier shows that the transporters are not always limited to the astrocytic endfeet in vivo. Our results suggest that the level of expression of monocarboxylic acid transporters MCT1 and MCT2 by cortical astrocytes in vivo is significantly lower than in vitro but that astrocytes in some other regions of the brain can express one or both proteins in significant amounts. PMID- 10794667 TI - KChAP as a chaperone for specific K(+) channels. AB - The concept of chaperones for K(+) channels is new. Recently, we discovered a novel molecular chaperone, KChAP, which increased total Kv2.1 protein and functional channels in Xenopus oocytes through a transient interaction with the Kv2.1 amino terminus. Here we report that KChAP is a chaperone for Kv1.3 and Kv4.3. KChAP increased the amplitude of Kv1.3 and Kv4.3 currents without affecting kinetics or voltage dependence, but had no such effect on Kv1.1, 1.2, 1.4, 1.5, 1.6, and 3.1 or Kir2.2, HERG, or KvLQT1. Although KChAP belongs to a family of proteins that interact with transcription factors, upregulation of channel currents was not blocked by the transcription inhibitor actinomycin D. A 98-amino acid fragment of KChAP binds to the channel and is indistinguishable from KChAP in its enhancement of Kv4.3 current and protein levels. Using a KChAP antibody, we have coimmunoprecipitated KChAP with Kv2.1 and Kv4.3 from heart. We propose that KChAP is a chaperone for specific Kv channels and may have this function in cardiomyocytes where Kv4.3 produces the transient outward current, I(to). PMID- 10794668 TI - NO increases permeability of cultured human cervical epithelia by cGMP-mediated increase in G-actin. AB - Human cervical epithelial cells express mRNA for the nitric oxide (NO) synthase (NOS) isoforms ecNOS, bNOS, and iNOS and release NO into the extracellular medium. N(G)-nitro-L-arginine methyl ester (L-NAME), an NOS inhibitor, and Hb, an NO scavenger, decreased paracellular permeability; in contrast, the NO donors sodium nitroprusside (SNP) and N-(ethoxycarbonyl)-3-(4-morpholinyl)sydnonimine increased paracellular permeability across cultured human cervical epithelia on filters, suggesting that NO increases cervical paracellular permeability. The objective of the study was to understand the mechanisms of NO action on cervical paracellular permeability. 8-Bromo-cGMP (8-BrcGMP) also increased permeability, and the effect was blocked by KT-5823 (a blocker of cGMP-dependent protein kinase), but not by LY-83583 (a blocker of guanylate cyclase). In contrast, LY 83583 and KT-5823 blocked the SNP-induced increase in permeability. Treatment with SNP increased cellular cGMP, and the effect was blocked by Hb and LY-83583, but not by KT-5823. Neither SNP nor 8-BrcGMP had modulated cervical cation selectivity. In contrast, both agents increased fluorescence from fura 2-loaded cells in the Ca(2+)-insensitive wavelengths, indicating that SNP and 8-BrcGMP stimulate a decrease in cell size and in the resistance of the lateral intercellular space. Neither SNP nor 8-BrcGMP had an effect on total cellular actin, but both agents increased the fraction of G-actin. Hb blocked the SNP induced increase in G-actin, and KT-5823 blocked the 8-BrcGMP-induced increase in G-actin. On the basis of these results, it is suggested that NO acts on guanylate cyclase and stimulates an increase in cGMP; cGMP, acting via cGMP-dependent protein kinase, shifts actin steady-state toward G-actin; this fragments the cytoskeleton and renders cells more sensitive to decreases in cell size and resistance of the lateral intercellular space and, hence, to increases in permeability. These results may be important for understanding NO regulation of transcervical paracellular permeability and secretion of cervical mucus in the woman. PMID- 10794670 TI - Probing the extracellular release site of the plasma membrane calcium pump. AB - The plasma membrane Ca(2+) pump is known to mediate Ca(2+)/H(+) exchange. Extracellular protons activated (45)Ca(2+) efflux from human red blood cells with a half-maximal inhibition constant of 2 nM when the intracellular pH was fixed. An increase in pH from 7.2 to 8.2 decreased the IC(50) for extracellular Ca(2+) from approximately 33 to approximately 6 mM. Changing the membrane potential by >54 mV had no effect on the IC(50) for extracellular Ca(2+). This argues against Ca(2+) release through a high-field access channel. Extracellular Ni(2+) inhibited Ca(2+) efflux with an IC(50) of 11 mM. Extracellular Cd(2+) inhibited with an IC(50) of 1. 5 mM, >10 times better than Ca(2+). The Cd(2+) IC(50) also decreased when the pH was raised from 7.1 to 8.2, consistent with Ca(2+), Cd(2+), and H(+) competing for the same site. The higher affinity for inhibition by Ni(2+) and Cd(2+) is consistent with a histidine or cysteine as part of the release site. The cysteine reagent 2-(trimethylammonium)ethyl methanethiosulfonate did not inhibit Ca(2+) efflux. Our results are consistent with the notion that the release site contains a histidine. PMID- 10794669 TI - Hypertonic perfusion inhibits intracellular Na and Ca accumulation in hypoxic myocardium. AB - Much evidence supports the view that hypoxic/ischemic injury is largely due to increased intracellular Ca concentration ([Ca](i)) resulting from 1) decreased intracellular pH (pH(i)), 2) stimulated Na/H exchange that increases Na uptake and thus intracellular Na (Na(i)), and 3) decreased Na gradient that decreases or reverses net Ca transport via Na/Ca exchange. The Na/H exchanger (NHE) is also stimulated by hypertonic solutions; however, hypertonic media may inhibit NHE's response to changes in pH(i) (Cala PM and Maldonado HM. J Gen Physiol 103: 1035 1054, 1994). Thus we tested the hypothesis that hypertonic perfusion attenuates acid-induced increases in Na(i) in myocardium and, thereby, decreases Ca(i) accumulation during hypoxia. Rabbit hearts were Langendorff perfused with HEPES buffered Krebs-Henseleit solution equilibrated with 100% O(2) or 100% N(2). Hypertonic perfusion began 5 min before hypoxia or normoxic acidification (NH(4)Cl washout). Na(i), [Ca](i), pH(i), and high-energy phosphates were measured by NMR. Control solutions were 295 mosM, and hypertonic solutions were adjusted to 305, 325, or 345 mosM by addition of NaCl or sucrose. During 60 min of hypoxia (295 mosM), Na(i) rose from 22+/-1 to 100+/-10 meq/kg dry wt while [Ca](i) rose from 347+/-11 to 1,306+/-89 nM. During hypertonic hypoxic perfusion (325 mosM), increases in Na(i) and [Ca](i) were reduced by 65 and 60%, respectively (P<0.05). Hypertonic perfusion also diminished Na uptake after normoxic acidification by 87% (P<0.05). The data are consistent with the hypothesis that mild hypertonic perfusion diminishes acid-induced Na accumulation and, thereby, decreases Na/Ca exchange-mediated Ca(i) accumulation during hypoxia. PMID- 10794671 TI - pH heterogeneity at intracellular and extracellular plasma membrane sites in HT29 C1 cell monolayers. AB - In the colonic mucosa, short-chain fatty acids change intracellular pH (pH(i)) and extracellular pH (pH(e)). In this report, confocal microscopy and dual emission ratio imaging of carboxyseminaphthorhodofluor-1 were used for direct evaluation of pH(i) and pH(e) in a simple model epithelium, HT29-C1 cells. Live cell imaging along the apical-to-basal axis of filter-grown cells allowed simultaneous measurement of pH in the aqueous environment near the apical membrane, the lateral membrane, and the basal membrane. Subapical cytoplasm reported the largest changes in pH(i) after isosmotic addition of 130 mM propionate or 30 mM NH(4)Cl. In resting cells and cells with an imposed acid load, lateral membranes had pH(i) values intermediate between the relatively acidic subapical region (pH 6.3-6.9) and the relatively alkaline basal pole of the cells (pH 7.4-7.1). Transcellular pH(i) gradients were diminished or eliminated during an induced alkaline load. Propionate differentially altered pH(e) near the apical membrane, in lateral intracellular spaces between adjacent cells, and near the basal membrane. Luminal or serosal propionate caused alkalinization of the cis compartment (where propionate was added) but acidification of the trans compartment only in response to luminal propionate. Addition of NH(4)Cl produced qualitatively opposite pH(e) excursions. The microscopic values of pH(i) and pH(e) can explain a portion of the selective activation of polarized Na/H exchangers observed in HT29-C1 cells in the presence of transepithelial propionate gradients. PMID- 10794672 TI - Apoptosis in human cultured trophoblasts is enhanced by hypoxia and diminished by epidermal growth factor. AB - Preeclampsia and fetal growth restriction are associated with placental hypoperfusion and villous hypoxia. The villous response to this environment includes diminished trophoblast differentiation and enhanced apoptosis. We tested the hypothesis that hypoxia induces apoptosis in cultured trophoblasts, and that epidermal growth factor (EGF), an enhancer of trophoblast differentiation, diminishes hypoxia-induced apoptosis. Trophoblasts isolated from placentas of term-uncomplicated human pregnancies were cultured up to 72 h in standard (PO(2) = 120 mm Hg) or hypoxic (PO(2) <15 mm Hg) conditions. Exposure to hypoxia for 24 h markedly enhanced trophoblast apoptosis as determined by DNA laddering, internucleosomal in situ DNA fragmentation, and histomorphology, as well as by the reversibility of the apoptotic process with a caspase inhibitor. Apoptosis was accompanied by increased expression of p53 and Bax and decreased expression of Bcl-2. Addition of EGF to cultured trophoblasts or exposure of more differentiated trophoblasts to hypoxia significantly lowered the level of apoptosis. We conclude that hypoxia enhances apoptosis in cultured trophoblasts by a mechanism that involves an increase in p53 and Bax expression. EGF and enhancement of cell differentiation protect against hypoxic-induced apoptosis. PMID- 10794673 TI - Ca(2+) regulates fluid shear-induced cytoskeletal reorganization and gene expression in osteoblasts. AB - Osteoblasts subjected to fluid shear increase the expression of the early response gene, c-fos, and the inducible isoform of cyclooxygenase, COX-2, two proteins linked to the anabolic response of bone to mechanical stimulation, in vivo. These increases in gene expression are dependent on shear-induced actin stress fiber formation. Here, we demonstrate that MC3T3-E1 osteoblast-like cells respond to shear with a rapid increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) that we postulate is important to subsequent cellular responses to shear. To test this hypothesis, MC3T3-E1 cells were grown on glass slides coated with fibronectin and subjected to laminar fluid flow (12 dyn/cm(2)). Before application of shear, cells were treated with two Ca(2+) channel inhibitors or various blockers of intracellular Ca(2+) release for 0. 5-1 h. Although gadolinium, a mechanosensitive channel blocker, significantly reduced the [Ca(2+)](i) response, neither gadolinium nor nifedipine, an L-type channel Ca(2+) channel blocker, were able to block shear-induced stress fiber formation and increase in c-fos and COX-2 in MC3T3-E1 cells. However, 1, 2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM, an intracellular Ca(2+) chelator, or thapsigargin, which empties intracellular Ca(2+) stores, completely inhibited stress fiber formation and c-fos/COX-2 production in sheared osteoblasts. Neomycin or U-73122 inhibition of phospholipase C, which mediates D myo-inositol 1,4,5-trisphosphate (IP(3))-induced intracellular Ca(2+) release, also completely suppressed actin reorganization and c-fos/COX-2 production. Pretreatment of MC3T3-E1 cells with U-73343, the inactive isoform of U-73122, did not inhibit these shear-induced responses. These results suggest that IP(3) mediated intracellular Ca(2+) release is required for modulating flow-induced responses in MC3T3-E1 cells. PMID- 10794674 TI - IP(3) receptors, IP(3) transients, and nucleus-associated Ca(2+) signals in cultured skeletal muscle. AB - Inositol 1,4,5-trisphosphate (IP(3)) receptors (IP(3)R) and ryanodine receptors (RyR) were localized in cultured rodent muscle fractions by binding of radiolabeled ligands (IP(3) and ryanodine), and IP(3)R were visualized in situ by fluorescence immunocytological techniques. Also explored was the effect of K(+) depolarization on IP(3) mass and Ca(2+) transients studied using a radio-receptor displacement assay and fluorescence imaging of intracellular fluo 3. RyR were located in a microsomal fraction; IP(3)R were preferentially found in the nuclear fraction. Fluorescence associated with anti-IP(3)R antibody was found in the region of the nuclear envelope and in a striated pattern in the sarcoplasmic areas. An increase in external K(+) affected membrane potential and produced an IP(3) transient. Rat myotubes displayed a fast-propagating Ca(2+) signal, corresponding to the excitation-contraction coupling transient and a much slower Ca(2+) wave. Both signals were triggered by high external K(+) and were independent of external Ca(2+). Slow waves were associated with cell nuclei and were propagated leaving "glowing" nuclei behind. Different roles are proposed for at least two types of Ca(2+) release channels, each mediating an intracellular signal in cultured skeletal muscle. PMID- 10794675 TI - Normalization of hyperosmotic-induced inositol uptake by renal and endothelial cells is regulated by NF-kappaB. AB - Hyperosmolarity is a stress factor that has been shown to cause an increase in the transcription of the Na(+)-dependent myo-inositol cotransporter (SMIT). However, regulation of the reversion of SMIT mRNA levels and transporter activity following removal of hyperosmotic stress is less understood. Previously we have shown that postinduction normalization of SMIT mRNA levels and myo-inositol accumulation following removal of hyperosmotic stress is inhibited by actinomycin D and cycloheximide, suggesting that normalization requires RNA transcription and protein synthesis. We now demonstrate that removal of hyperosmotic stress causes an activation of the transcription factor NF-kappaB in renal and endothelial cells. Inhibiting NF-kappaB activation with pyrrolidine dithiocarbamate (PD) blocks the normalization of SMIT mRNA levels and myo-inositol accumulation on removal of the cells from hyperosmotic medium. These studies demonstrate that the downregulation of the myo-inositol transporter following reversal of hyperosmotic induction is regulated via the activation of NF-kappaB. PMID- 10794676 TI - Structure, function, and genomic organization of human Na(+)-dependent high affinity dicarboxylate transporter. AB - We have cloned and functionally characterized the human Na(+)-dependent high affinity dicarboxylate transporter (hNaDC3) from placenta. The hNaDC3 cDNA codes for a protein of 602 amino acids with 12 transmembrane domains. When expressed in mammalian cells, the cloned transporter mediates the transport of succinate in the presence of Na(+) [concentration of substrate necessary for half-maximal transport (K(t)) for succinate = 20+/-1 microM]. Dimethylsuccinate also interacts with hNaDC3. The Na(+)-to-succinate stoichiometry is 3:1 and concentration of Na(+) necessary for half-maximal transport (K(Na(+))(0.5)) is 49+/-1 mM as determined by uptake studies with radiolabeled succinate. When expressed in Xenopus laevis oocytes, hNaDC3 induces Na(+)-dependent inward currents in the presence of succinate and dimethylsuccinate. At a membrane potential of -50 mV, K(Suc)(0.5) is 102+/-20 microM and K(Na(+))(0.5) is 22+/-4 mM as determined by the electrophysiological approach. Simultaneous measurements of succinate-evoked charge transfer and radiolabeled succinate uptake in hNaDC3-expressing oocytes indicate a charge-to-succinate ratio of 1:1 for the transport process, suggesting a Na(+)-to-succinate stoichiometry of 3:1. pH titration of citrate-induced currents shows that hNaDC3 accepts preferentially the divalent anionic form of citrate as a substrate. Li(+) inhibits succinate-induced currents in the presence of Na(+). Functional analysis of rat-human and human-rat NaDC3 chimeric transporters indicates that the catalytic domain of the transporter lies in the carboxy-terminal half of the protein. The human NaDC3 gene is located on chromosome 20q12-13.1, as evidenced by fluorescent in situ hybridization. The gene is >80 kbp long and consists of 13 exons and 12 introns. PMID- 10794677 TI - Pharmacology and modulation of K(ATP) channels by protein kinase C and phosphatases in gallbladder smooth muscle. AB - ATP-sensitive K(+) (K(ATP)) channels exhibit pharmacological diversity, which is critical for the development of novel therapeutic agents. We have characterized K(ATP) channels in gallbladder smooth muscle to determine how their pharmacological properties compare to K(ATP) channels in other types of smooth muscle. K(ATP) currents were measured in myocytes isolated from gallbladder and mesenteric artery. The potencies of pinacidil, diazoxide, and glibenclamide were similar in gallbladder and vascular smooth muscle, suggesting that the regions of the channel conferring sensitivity to these agents are conserved among smooth muscle types. Activators of protein kinase C (PKC), however, were less effective at inhibiting K(ATP) currents in myocytes from gallbladder than mesenteric artery. The phosphatase inhibitor okadaic acid increased the efficacy of PKC activators and revealed ongoing basal activation of K(ATP) channels by protein kinase A in gallbladder. These results suggest that phosphatases and basal kinase activity play an important role in controlling K(ATP) channel activity. PMID- 10794678 TI - Impermeability of the GIRK2 weaver channel to divalent cations. AB - A single amino acid mutation (G156S) in the putative pore-forming region of the G protein-sensitive, inwardly rectifying K(+) channel subunit, GIRK2, renders the conductance constitutively active and nonselective for monovalent cations. The mutant channel subunit (GIRK2wv) causes the pleiotropic weaver disease in mice, which is characterized by the selective vulnerability of cerebellar granule cells and Purkinje cells, as well as dopaminergic neurons in the mesencephalon, to cell death. It has been proposed that divalent cation permeability through constitutively active GIRK2wv channels contributes to a rise in internal calcium in the GIRK2wv-expressing neurons, eventually leading to cell death. We carried out comparative studies of recombinant GIRK2wv channels expressed in Xenopus oocytes and COS-7 cells to determine the magnitude and relative permeability of the GIRK2wv conductance to Ca(2+). Data from these studies demonstrate that the properties of the expressed current differ in the two systems and that when recombinant GIRK2wv is expressed in mammalian cells it is impermeable to Ca(2+). PMID- 10794679 TI - Kinase regulation of hENaC mediated through a region in the COOH-terminal portion of the alpha-subunit. AB - In an effort to gain insight into how kinases might regulate epithelial Na(+) channel (ENaC) activity, we expressed human ENaC (hENaC) in Xenopus oocytes and examined the effect of agents that modulate the activity of some kinases. Activation of protein kinase C (PKC) by phorbol ester increased the activity of ENaC, but only in oocytes with a baseline current of <2,000 nA. Inhibitors of protein kinases produced varying effects. Chelerythrine, an inhibitor of PKC, produced a significant inhibition of ENaC current, but calphostin C, another PKC inhibitor, had no effect. The PKA/protein kinase G inhibitor H-8 had no effect, whereas the p38 mitogen-activated protein kinase inhibitor, SB-203580 had a significant inhibitory effect. Staurosporine, a nonspecific kinase inhibitor, was the most potent tested. It inhibited ENaC currents in both oocytes and in M-1 cells, a model for the collecting duct. Site-directed mutagenesis revealed that the staurosporine effect did not require an intact COOH terminus of either the beta- or gamma-hENaC subunit. However, an intact COOH terminus of the alpha subunit was required for this effect. These results suggest that an integrated kinase network regulates ENaC activity through an action that requires a portion of the alpha-subunit. PMID- 10794680 TI - LabPatch, an acquisition and analysis program for patch-clamp electrophysiology. AB - An acquisition and analysis program, "LabPatch," has been developed for use in patch-clamp research. LabPatch controls any patch-clamp amplifier, acquires and records data, runs voltage protocols, plots and analyzes data, and connects to spreadsheet and database programs. Controls within LabPatch are grouped by function on one screen, much like an oscilloscope front panel. The software is mouse driven, so that the user need only point and click. Finally, the ability to copy data to other programs running in Windows 95/98, and the ability to keep track of experiments using a database, make LabPatch extremely versatile. The system requirements include Windows 95/98, at least a 100-MHz processor and 16 MB RAM, a data acquisition card, digital-to-analog converter, and a patch-clamp amplifier. LabPatch is available free of charge at http://www.fhs.mcmaster.ca/huizinga/. PMID- 10794681 TI - Corticotropin releasing factor (CRF) receptor modulators: progress and opportunities for new therapeutic agents. PMID- 10794682 TI - N-[[(5-methyl-3-phenylisoxazol-4-yl)-phenyl]sulfonyl]propanamide, sodium salt, parecoxib sodium: A potent and selective inhibitor of COX-2 for parenteral administration. PMID- 10794683 TI - Potent derivatives of glucagon-like peptide-1 with pharmacokinetic properties suitable for once daily administration. AB - A series of very potent derivatives of the 30-amino acid peptide hormone glucagon like peptide-1 (GLP-1) is described. The compounds were all derivatized with fatty acids in order to protract their action by facilitating binding to serum albumin. GLP-1 had a potency (EC(50)) of 55 pM for the cloned human GLP-1 receptor. Many of the compounds had similar or even higher potencies, despite quite large substituents. All compounds derivatized with fatty acids equal to or longer than 12 carbon atoms were very protracted compared to GLP-1 and thus seem suitable for once daily administration to type 2 diabetic patients. A structure activity relationship was obtained. GLP-1 could be derivatized with linear fatty acids up to the length of 16 carbon atoms, sometimes longer, almost anywhere in the C-terminal part without considerable loss of potency. Derivatization with two fatty acid substituents led to a considerable loss of potency. A structure activity relationship on derivatization of specific amino acids generally was obtained. It was found that the longer the fatty acid, the more potency was lost. Simultaneous modification of the N-terminus (in order to obtain better metabolic stability) interfered with fatty acid derivatization and led to loss of potency. PMID- 10794684 TI - Substituted benzamide inhibitors of human rhinovirus 3C protease: structure-based design, synthesis, and biological evaluation. AB - A series of nonpeptide benzamide-containing inhibitors of human rhinovirus (HRV) 3C protease was identified using structure-based design. The design, synthesis, and biological evaluation of these inhibitors are reported. A Michael acceptor was combined with a benzamide core mimicking the P1 recognition element of the natural 3CP substrate. alpha,beta-Unsaturated cinnamate esters irreversibly inhibited the 3CP and displayed antiviral activity (EC(50) 0.60 microM, HRV-16 infected H1-HeLa cells). On the basis of cocrystal structure information, a library of substituted benzamide derivatives was prepared using parallel synthesis on solid support. A 1.9 A cocrystal structure of a benzamide inhibitor in complex with the 3CP revealed a binding mode similar to that initially modeled wherein covalent attachment of the nucleophilic cysteine residue is observed. Unsaturated ketones displayed potent reversible inhibition but were inactive in the cellular antiviral assay and were found to react with nucleophilic thiols such as DTT. PMID- 10794685 TI - Molecular determinants of MAO selectivity in a series of indolylmethylamine derivatives: biological activities, 3D-QSAR/CoMFA analysis, and computational simulation of ligand recognition. AB - A series of indolylmethylamine derivatives were assayed toward MAO-A and MAO-B inhibition. The K(i) values of these compounds are in the range from 0.8 to >10(6) nM for MAO-A or from 0.75 to 476000 nM for MAO-B. The most selective MAO-A or MAO-B inhibitors elicit a ratio of K(i) in the order of 1500 or 1000, respectively. Comparison of MAO-A and MAO-B CoMFA models showed that both the steric and electrostatic properties at the 5 position of the indole ring are determinant for MAO selectivity. Computational simulations of the complex between this part of the ligand and Phe-208 of MAO-A or Ile-199 of MAO-B, experimentally identified as responsible for substrate selectivity, allowed us to further characterize the nature of these enzyme-inhibitor interactions. PMID- 10794686 TI - Probing the conformation of the sugar transport inhibitor phlorizin by 2D-NMR, molecular dynamics studies, and pharmacophore analysis. AB - Sodium/D-glucose cotransport, one of the prototypes for sodium gradient-driven symport systems in kidney and intestine, is known to be inhibited by aromatic and aliphatic glucosides (Diedrich, D. F. Biochim. Biophys. Acta 1963, 71, 688-700; Diedrich, D. F. Arch. Biochem. Biophys. 1966, 117, 248-256; Kipp, H.; et al. Biochim. Biophys. Acta 1996, 1282, 124-130; Ramaswamy, K.; et al. Biochim. Biophys. Acta 1976, 433, 32-38). The conformation in which the most potent inhibitor, phlorizin, interacts with the transport protein was investigated with different approaches. Phlorizin consists of the glucose moiety and two aromatic rings (A and B) joined by an alkyl spacer. First the interaction of these various parts of the molecule was determined by two-dimensional (2D) solution NMR. From the 2D-NOESY (nuclear Overhauser effect) measurements spatial distances (up to 5 A) between various interacting H atoms could be detected. Using these values as distance constraints, conformations of phlorizin were calculated and analyzed by the valence force-field method. As a result, a set of conformations could be obtained. The most probable phlorizin conformation shows a nearly perpendicular arrangement of the two aromatic rings (A and B) with the ring B situated above the sugar ring. A very similar conformation could be found by using molecular dynamics simulations when water was chosen as the solvent. This phlorizin conformation in aqueous solution then served as a template for conformational analysis of various phlorizin derivatives. The resulting conformations of derivatives were taken as input to establish a pharmacophore model using the DISCO calculation. As a result, the essential elements of phlorizin for interaction with its binding pocket could be deduced: namely hydrogen bonding via hydroxyl groups of the pyranoside at C(2), C(3), C(4), and C(6) and at C(4) and C(6) of aromatic ring A and hydrophobic interactions via the pyranoside ring and aromatic ring A. Finally, from these conformational features of the pharmacophore the dimension of the phlorizin binding site on the sodium/D-glucose cotransporter was estimated to be 17 x 10 x 7 A(3). PMID- 10794687 TI - 2-amino-2-oxazolines as subtype selective alpha(2) adrenoceptor agonists. AB - Cyclohexylamino oxazoline 1 (AGN 190837), an analogue of 2 (Bay a6781), is a potent alpha(2) adrenoceptor agonist. On the basis of a design generated by receptor-ligand modeling, a number of cyclohexyl and norbornyl analogues were synthesized wherein the propyl group of 1 was replaced by phenylalkyl subsituents. This resulted in compound 6 being an alpha(2c) selective agonist, as well as 7 and 9 being alpha(2a)/alpha(2c) selective. PMID- 10794688 TI - New proline mimetics: synthesis of thrombin inhibitors incorporating cyclopentane and cyclopentenedicarboxylic acid templates in the P2 position. Binding conformation investigated by X-ray crystallography. AB - With the aim to prepare nonpeptidic thrombin inhibitors, the amino acids of the thrombin-inhibiting tripeptide chain D-Phe-Pro-Arg were replaced with isosteres. Arg was replaced with the more rigid P1 truncated p-amidinobenzylamine (Pab), Pro with either cyclopentane-1, 2-dicarboxylic acid or cyclopentene-1,5-dicarboxylic acid, and D-Phe with a series of readily available lipophilic amines. One of the most potent compounds (25, pIC(50) = 6.01) in these series was cocrystallized with thrombin where the X-ray crystal structure provide insight to the structure activity relationship (SAR). PMID- 10794689 TI - Design and synthesis of conformationally constrained glucagon analogues. AB - Glucagon was systematically modified by forming lactam bridges within the central region of the molecule to give conformationally constrained cyclic analogues. Six cyclic glucagon analogues have been designed and synthesized. They are c[Asp(9),Lys(12)][Lys(17,18), Glu(21)]glucagon-NH(2) (1), c[Asp(9),Lys(12)]glucagon-NH(2) (2), c[Lys(12),Asp(15)]glucagon-NH(2) (3), c[Asp(15), Lys(18)]glucagon-NH(2) (4), [Lys(17)-c[Lys(18), Glu(21)]glucagon-NH(2) (5), and c[Lys(12),Asp(21)]glucagon-NH(2) (6). The receptor binding potencies and receptor second messenger activities were determined by radio-receptor binding assays and adenylate cyclase assays, respectively, using rat liver plasma membranes. Most interestingly, analogues 1, 2, 3, and 4 were antagonists of glucagon stimulated adenylate cyclase activity, whereas analogues 5 and 6 were partial agonists in the functional assay. All of the cyclic analogues were found to have reduced binding potencies relative to glucagon. The structural features that might be responsible for these effects were studied using circular dichroism spectroscopy and molecular modeling. These results demonstrated the significant modulations of both receptor binding affinity and transduction (adenylate cyclase activity) that can accompany regional conformational constraints even in larger polypeptide ligands. These studies suggest that the entire molecular conformation, including the flexible middle portion, is important for molecular recognition and transduction at the hepatic glucagon receptor. PMID- 10794690 TI - Toward general methods of targeted library design: topomer shape similarity searching with diverse structures as queries. AB - A promising strategy for selecting synthetic targets is similarity-based searching of very large "virtual libraries", which comprise all structures accessible by linking two or three commercially available building blocks with combinatorial syntheses. To assess the general applicability of this strategy, leading structures taken from each of 34 recent medicinal chemistry publications were used as queries to search a virtual library containing 2.6 x 10(13) products from seven reactions, using a topomer shape similarity metric. Eighty-five percent of these searches succeeded, by yielding, with a search radius no greater than 120 topomer shape units, either at least 400 hits or hits from at least six sublibraries. From these 34 sets of search results, 122 representative structures were selected, illustrating potential "lead hops", or otherwise novel structures. Overall shape similarity to the query structure was confirmed for up to 95% of these representative structures, according to FLEXS, an algorithmically distinct program. Experimentally, there were 28 structures among those reported in the 34 query publications that were identified within the virtual library. Among these, the frequency of high activity was 87% for the 16 structures whose similarity to their query was 90 topomer units or less, compared to a frequency of 50% for the other 12 structures. PMID- 10794691 TI - Solution structures in SDS micelles and functional activity at the bullfrog substance P receptor of ranatachykinin peptides. AB - A set of novel tachykinin-like peptides has been isolated from bullfrog brain and gut. These compounds, ranatachykinin A (RTKA), ranatachykinin B (RTKB), and ranatachykinin C (RTKC), were named for their source, Rana catesbeiana, and their homology to the tachykinin peptide family. We present the first report of the micelle-bound structures and pharmacological actions of the RTKs. Generation of three-dimensional structures of the RTKs in a membrane-model environment using (1)H NMR chemical shift assignments, two-dimensional NMR techniques, and molecular dynamics and simulated annealing procedures allowed for the determination of possible prebinding ligand conformations. RTKA, RTKB, and RTKC were determined to be helical from the midregion to the C-terminus (residues 4 10), with a large degree of flexibility in the N-terminus and minor dynamic fraying at the end of the C-terminus. The pharmacological effects of the RTKs were studied by measuring the elevation of intracellular Ca(2+) in Chinese hamster ovarian cells stably transfected with the bullfrog substance P receptor (bfSPR). All of the RTKs tested elicited Ca(2+) elevations with a rank order of maximal effect of RTKA >/= SP > RTKC >/= RTKB. A high concentration (1 microM) of the neuropeptides produced varying degrees of desensitization to a subsequent challenge with the same or different peptide, while a low concentration (1 pM) produced sensitization at the bfSPR. Our data suggest differences in amino acid side chains and their charged states at the C-terminal sequence or differences in secondary structure at the N-terminus, which do not overlap according to the findings in this paper, may explain the differing degree and type of receptor activation seen at the bfSPR. PMID- 10794692 TI - Characterization of the binding site of the histamine H(3) receptor. 2. Synthesis, in vitro pharmacology, and QSAR of a series of monosubstituted benzyl analogues of thioperamide. AB - A series of monosubstituted benzyl analogues of the histamine H(3) receptor antagonist thioperamide were synthesized and evaluated for their histamine H(3) receptor activity on the guinea pig jejunum. Incorporation of Cl, Br, and I at the ortho position of the benzyl moiety led to an increase of the pA(2) value, whereas the same substituents at the para position led to a decrease. However, a fluorine substituent gave a strong decrease in pA(2), regardless of the position. Molecular modeling revealed a QSAR with a correlation (r = 0.93) between the pA(2) and the dihedral angle between the thiourea and the benzyl moiety and the calculated electron density on the substituted carbon atom. To verify whether this QSAR model had a predictive value, the ortho tert-butyl and methyl analogues were synthesized and evaluated. Indeed it was shown that the predicted pA(2) values of these two compounds were in accordance with the measured pA(2) values. PMID- 10794693 TI - Screening of new antioxidant molecules using flow cytometry. AB - We present a flow cytometry technique to evaluate the antioxidative properties of molecules on living cells, using a stable murine-murine hybridoma (Mark 3) cell line routinely cultured. Using this technique, intracellular superoxide anions and peroxides were evaluated with dihydrorhodamine (DHR-123) and dichlorofluorescein diacetate (DCFH-DA), respectively. When cells were first incubated for 10 min with either H(2)O(2) or the xanthine (X)/xanthine oxidase (XO) system, this flow cytometric technique was capable of evaluating the oxidative stress on cells. Twenty-one new analogues of ellipticine were synthesized and tested for their antioxidative properties compared to vitamin E and Ebselen used as references. A good statistical reflection of the antioxidative activities of these molecules was achieved by analyzing 35 000 cells in each experiment. Among them, the selenated molecule 18 was found to be 10 times more active than Ebselen but 10 000 times less active than vitamin E. Moreover, eight compounds showed glutathione peroxidase-like activities. PMID- 10794695 TI - Nuclear receptor-DNA binding specificity: A COMBINE and Free-Wilson QSAR analysis. AB - Specific binding of transcription factors to DNA is crucial for gene regulation. We derived models for the binding specificity of transcription factors of the nuclear receptor family to DNA using two QSAR methods: a Free-Wilson-like method and COMparative BINding Energy (COMBINE) analysis. The analysis is based on experimental data for the interaction of 20 mutant glucocorticoid receptor DNA binding domains with 16 different response elements in a total of 320 complexes (Zilliacus, J.; Wright, A. P.; Carlstedt-Duke, J.; Nilsson, L.; Gustafsson, J. A. Proteins 1995, 21, 57-67). The predictive abilities of the models obtained by the two methods are similar. The COMBINE analysis indicates that the most important properties for determining binding specificity for this dataset are the changes upon binding of the solvation free energies of the bases that are mutated in the dataset and the electrostatic interactions of the mutated nucleotides with certain charged amino acids. Further important descriptors are the changes of solvation free energy and surface area of the side chain of the mutated residue. It is clear, however, that there are additional features important for the specificity of binding that are not included in the models, such as differences in interfacial hydration of the complexes. PMID- 10794694 TI - Design of a Gag pentapeptide analogue that binds human cyclophilin A more efficiently than the entire capsid protein: new insights for the development of novel anti-HIV-1 drugs. AB - Cyclophilin A (hCyp-18), a ubiquitous cytoplasmic peptidyl-prolyl cis/trans isomerase (PPIase), orchestrates HIV-1 core packaging. hCyp-18, incorporated into the virion, enables core uncoating and RNA release and consequently plays a critical role in the viral replication process. hCyp-18 specifically interacts with a single exposed loop of the Gag polyprotein capsid domain via a network of nine hydrogen bonds which mainly implicates a 7-mer fragment of the loop. As previously reported, the corresponding linear heptapeptide Ac-Val-His-Ala-Gly-Pro Ile-Ala-NH(2) (2) binds to hCyp-18 with a low affinity (IC(50) = 850 +/- 220 microM) but a potentially useful selectivity for hCyp-18 relative to hFKBP-12, another abundant PPIase. On the basis of X-ray structures of Gag fragments:hCyp 18 complexes, we generated a series of modified peptides in order to probe the determinants of the interaction and hence to select a peptidic ligand displaying a higher affinity than the capsid domain of Gag. We synthesized a series of heptapeptides to test the energetic contribution of amino acids besides the Gly Pro moiety. In particular the importance of the histidine residue for the interaction was underscored. We also investigated the influence of N- and C terminal modifications. Hexapeptides containing either deaminovaline (Dav) in place of the N-terminal valine or substitution of the C-terminal alanine amide with a benzylamide group displayed increased affinities. Combination of both modifications gave the most potent competitor Dav-His-Ala-Gly-Pro-Ile-NHBn (28) which has a higher affinity for hCyp-18 (K(d) = 3 +/- 0.5 microM) than the entire capsid protein (K(d) = 16 +/- 4 microM) and a very low affinity for hFKBP-12. Some of our results strongly suggest that the title compound is not a substrate of hCyp-18 and interacts preferentially in the trans conformation. PMID- 10794696 TI - Protease inhibitors: synthesis and QSAR study of novel classes of nonbasic thrombin inhibitors incorporating sulfonylguanidine and O-methylsulfonylisourea moieties at P1. AB - Using benzamidine as a lead molecule, two series of alkyl/aralkyl/arylsulfonylguanidines/sulfonyl-O-methylisoureas+ ++ have been prepared and assayed as inhibitors of two serine proteases, thrombin and trypsin. The study showed that sulfaguanidine and its corresponding O-methylisourea derivative possess moderate but intrinsically selective thrombin inhibitory properties, with K(I)'s around 100 nM against thrombin and 1350-1500 nM against trypsin. Further elaboration of these two molecules afforded compounds that inhibited thrombin with K(I)'s in the range of 12-50 nM, whereas affinity for trypsin remained relatively low. Such compounds were obtained by attaching benzyloxycarbonyl- or 4-toluenesulfonylureido-protected amino acids (such as L- and D-Phe or L-Pro) or dipeptides (such as Phe-Pro, Gly-His, beta-Ala-His, or Pro Gly) to the two leads mentioned above, sulfaguanidine and 4-aminobenzenesulfonyl O-methylisourea. Thus, the present study proposes two novel approaches for the preparation of high-affinity, specific thrombin inhibitors: two novel S1 anchoring moieties in the already large family of arginine/amidine-based inhibitors and novel peptidomimetic scaffolds obtained by incorporating tosylureido amino acids in the hydrophobic binding site(s). The first one is important for obtaining bioavailable thrombin inhibitors, devoid of the high basicity of the commonly used arginine/amidine-based inhibitors, whereas the second one may lead to improved water solubility of such compounds due to facilitated metal (sodium) salts formation (at the relatively acidic SO(2)NHCO protons) as well as increased stability at hydrolysis (in vivo). A QSAR study also explained the activity in terms of global properties of the molecules, electronic properties of the sulfonylguanidine/sulfonylisourea moiety, and novel descriptors, the frontier orbital phase angles (FOPA), that account for the directions of the nodes in the pi orbitals in the aromatic portion of those of the drugs in which the sulfonyl group was bound to a benzene ring. For thrombin inhibition, the size of the molecule was the dominant influence, while for trypsin inhibition the FOPA was the principal determinant of activity. The dependence of activity on the FOPA variables is perhaps the clearest example of a quantum effect in pharmacology and suggests a promising new tool for drug design. PMID- 10794697 TI - Multivariate data analysis using D-optimal designs, partial least squares, and response surface modeling: A directional approach for the analysis of farnesyltransferase inhibitors. AB - We have investigated the combined use of partial least squares (PLS) and statistical design principles in principal property space (PP-space), derived from principal component analysis (PCA), to analyze farnesyltransferase inhibitors in order to identify "activity trends" (an approach we call a "directional" approach) and quantitative structure-activity relationships (QSAR) for a congeneric series of inhibitors: the benzo[f]perhydroisoindole (BPHI) series. Trends observed in the PCA showed that the descriptors used were relevant to describe our structural data set by clearly identifying two well-defined structural subclasses of inhibitors. D-Optimal design techniques allowed us to define a training set for PLS study in PP-space. Models were derived for each biological assay under evaluation: the in vitro Ki-Ras and cellular HCT116 tests. Each of these assay-based sets was subdivided once more into two subsets according to two structural classes in this BPHI series as revealed by the PCA model. The response surface modeling (RSM) methodology was used for each subset, and the corresponding RSM plots helped us identify "activity trends" exploited to guide further analogue design. For more precise activity predictions more refined PLS models on constrained PP-spaces were developed for each subset. This approach was validated with predicted sets and demonstrates that useful information can be extracted from just a few very informative and representative compounds. Finally, we also showed the potential use of such a strategy at an early stage of an optimization process to extract the first "activity trends" that might support decision making and guide medicinal chemists in the initial design of new analogues and/or lead followup libraries. PMID- 10794698 TI - Synthesis and cytostatic activity of substituted 6-phenylpurine bases and nucleosides: application of the Suzuki-Miyaura cross-coupling reactions of 6 chloropurine derivatives with phenylboronic acids. AB - The Suzuki-Miyaura reaction of protected 6-chloropurine and 2-amino-6 chloropurine bases and nucleosides with substituted phenylboronic acids led to the corresponding protected 6-(substituted phenyl)purine derivatives 6-9. Their deprotection yielded a series of substituted 6-phenylpurine bases and nucleosides 10-13. Significant cytostatic activity (IC(50) 0.25-20 micromol/L) in CCRF-CEM, HeLa, and L1210 cell lines was found for several 6-(4-X-substituted phenyl)purine ribonucleosides 12 (X = H, F, Cl, and OR), while the 6-phenylpurine and 2-amino-6 phenylpurine bases 10 and 11, as well as 2-amino-6-phenylpurine ribosides 13, were entirely inactive against these cell lines. PMID- 10794699 TI - Design and synthesis of aryloxyethyl thiocyanate derivatives as potent inhibitors of Trypanosoma cruzi proliferation. AB - As a part of our project directed at the search of new chemotherapeutic agents against American trypanosomiasis (Chagas' disease), several drugs possessing the 4-phenoxyphenoxy skeleton and other closely related structures employing the thiocyanate moiety as polar end group were designed, synthesized, and evaluated as antiproliferative agents against Trypanosoma cruzi, the parasite responsible for this disease. These thiocyanate analogues were envisioned bearing in mind the potent activity shown by 4-phenoxyphenoxyethyl thiocyanate (compound 8) taken as lead drug. This compound had previously proved to be an extremely active growth inhibitor against T. cruzi with IC(50) values ranging from the very low micromolar level in epimastigotes to the low nanomolar level in the intracellular form of the parasite. Of the designed compounds, the ethyl thiocyanate drugs connected to nonpolar skeletons, namely, arylthio, 2,4-dichlorophenoxy, ortho substituted aryloxy, and 2-methyl-4-phenoxyphenoxy (compounds 15, 34, 47, 52, 72, respectively), were shown to be very potent antireplicative agents against T. cruzi. On the other hand, conformationally restricted analogues as well as branched derivatives at the aliphatic side chain were shown to be moderately active against T. cruzi growth. The biological activity of drugs bearing the thiocyanate group correlated quite well with the activity exhibited by their normal precursors, the tetrahydropyranyl ether derivatives, when bonded to the same nonpolar skeleton. Compounds having the tetrahydropyranyl moeity as polar end were proportionally much less active than sulfur-containing derivatives in all cases. Drugs 47 and 72 also resulted to be very active against the amastigote form of the parasite growing in myoblasts; however, they were slightly less active than the lead drug 8. On the other hand, compounds 34 and 52 were almost devoid of activity against myoblasts. Surprisingly, the dithio derivative 15 was toxic for myoblasts. PMID- 10794700 TI - 1-imidazolyl(alkyl)-substituted di- and tetrahydroquinolines and analogues: syntheses and evaluation of dual inhibitors of thromboxane A(2) synthase and aromatase. AB - A series of 1-imidazolyl(alkyl)-substituted quinoline, isoquinoline, naphthalene, benzo[b]furan, and benzo[b]thiophene derivatives was synthesized as dual inhibitors of thromboxane A(2) synthase (P450 TxA(2)) and aromatase (P450 arom). Dual inhibition of these enzymes could be a novel strategy for the treatment of mammary tumors and the prophylaxis of metastases. The most potent dual inhibitors, 5-(2-imidazol-1-ylethyl)-7,8-dihydroquinoline (31) (P450 TxA(2): IC(50) = 0.29 microM; P450 arom: IC(50) = 0.50 microM) and its 5, 6-saturated analogue 30 (P450 TxA(2): IC(50) = 0.68 microM; P450 arom: IC(50) = 0.38 microM), showed a stronger inhibition of both target enzymes than the reference compounds (dazoxiben: IC(50) = 1.1 microM; aminoglutethimide: IC(50) = 18.5 microM). For the determination of the in vivo activity, the influence of selected compounds on serum TxB(2) concentration was examined in rats. Compound 30 (8.5 mg/kg body weight) led to a reduction of the TxB(2) serum level of 78%, 71%, and 51% after 3, 5, and 8 h, respectively (dazoxiben: 60%, 34%, and 36%). Selectivity was studied toward some enzymes of the steroidogenic and eicosanoid pathways. P450 17 was inhibited by selected compounds only at high concentrations. Compound 30 inhibited P450 scc by 13% (25 microM). Compound 31 did not affect cyclooxygenase and lipoxygenase. PMID- 10794701 TI - Structural determinants of opioid activity in the orvinols and related structures: ethers of orvinol and isoorvinol. AB - A series of ethers of orvinol and isoorvinol has been prepared and evaluated in opioid receptor binding and in vitro functional assays. The most striking finding was the very large difference in kappa-opioid receptor activity between the diastereomeric ethyl ethers: 46-fold in binding, 150-fold in GPI, and 900-fold in the [(35)S]GTPgammaS assay in favor of the (R)-diastereomer. Additionally in the (R)-series there was a 700-fold increase in kappa-agonist potency in the [(35)S]GTPgammaS assay when OEt was replaced by OBn. The data can be explained in a triple binding site model: an H-bonding site, a lipophilic site, and an inhibitory site with which the 20-Me group in the (S)-ethers may interact. It appears that kappa-agonist binding of the orvinols avoids the inhibitory site in the intramolecular H-bonded conformation. PMID- 10794702 TI - Protease inhibitors: synthesis of potent bacterial collagenase and matrix metalloproteinase inhibitors incorporating N-4-nitrobenzylsulfonylglycine hydroxamate moieties. AB - A series of compounds was prepared by reaction of alkyl/arylsulfonyl halides with N-4-nitrobenzylglycine, followed by conversion of the COOH to the CONHOH group, with hydroxylamine in the presence of carbodiimides. Other structurally related compounds were obtained by reaction of N-4-nitrobenzylglycine with aryl isocyanates, arylsulfonyl isocyanates, or benzoyl isothiocyanate, followed by the similar conversion of the COOH into the CONHOH moiety. Another subseries of derivatives was prepared from sulfanilyl- or metanilyl-4-nitrobenzylglycine by reaction with arylsulfonyl isocyanates, followed by conversion of the COOH to the hydroxamate moiety. The new compounds were assayed as inhibitors of four matrix metalloproteinases (MMPs), MMP-1, MMP-2, MMP-8, and MMP-9, and of the Clostridium histolyticum collagenase (ChC). Some of the prepared hydroxamate derivatives proved to be very effective collagenase/gelatinase inhibitors, depending on the substitution pattern at the sulfonamido moiety. Substitutions leading to best inhibitors of MMP-1, a short pocket enzyme, were those involving pentafluorophenylsulfonyl or 3-trifluoromethylphenylsulfonyl moieties at P(1') (K(I)'s of 3-5 nM). For MMP-2, MMP-8, and MMP-9 (deep-pocket enzymes), best inhibitors were especially those containing long perfluoroalkylsulfonyl and substituted-arylsulfonyl moieties, such as pentafluorophenylsulfonyl, 3- and 4 protected-aminophenylsulfonyl, 3- and 4-carboxyphenylsulfonyl, arylsulfonylureido, or arylsulfonylureidosulfanilyl/metanilyl moieties, at P(1'). Bulkier groups in this position, such as 1- and 2-naphthyl, substituted-naphthyl, or quinolin-8-yl moieties among others, led to less effective MMP/ChC inhibitors. Best ChC inhibitors were again those containing pentafluorophenylsulfonyl or 3- and 4-protected-aminophenylsulfonyl P(1') anchoring groups, suggesting that this protease is also a short-pocket wider-neck one (more similar to MMP-1). This study also proves that the 4-nitrobenzyl moiety is an efficient P(2') anchoring moiety and that sulfonylureido, ureido, or carboxythioureido substitutions at P(1') are also tolerated for obtaining potent sulfonylated amino acid hydroxamate like MMP/ChC inhibitors. PMID- 10794703 TI - Novel synthetic oleanane and ursane triterpenoids with various enone functionalities in ring A as inhibitors of nitric oxide production in mouse macrophages. AB - We initially randomly synthesized about 60 oleanane and ursane triterpenoids as potential anti-inflammatory and cancer chemopreventive agents. Preliminary screening of these derivatives for inhibition of production of nitric oxide induced by interferon-gamma in mouse macrophages revealed that 3-oxooleana-1, 12 dien-28-oic acid (B-15) showed significant activity (IC(50) = 5.6 microM). On the basis of the structure of B-15, 19 novel olean- and urs-12-ene triterpenoids with a 1-en-3-one functionality having a substituent at C-2 in ring A have been designed and synthesized. Among them, 3-oxooleana-1,12-diene derivatives with carboxyl, methoxycarbonyl, and nitrile groups at C-2 showed higher activity than the lead compound B-15. In particular, 2-carboxy-3-oxooleana-1, 12-dien-28-oic acid (3) had the highest activity (IC(50) = 0.07 microM) in this group of triterpenoids. The potency of 3 was similar to that of hydrocortisone (IC(50) = 0.01 microM), although 3 does not act through the glucocorticoid receptor. Interesting structure-activity relationships of these novel synthetic triterpenoids are also discussed. PMID- 10794704 TI - Design and synthesis of trans-N-[4-[2-(6-cyano-1,2,3, 4-tetrahydroisoquinolin-2 yl)ethyl]cyclohexyl]-4-quinolinecarboxamide (SB-277011): A potent and selective dopamine D(3) receptor antagonist with high oral bioavailability and CNS penetration in the rat. AB - A selective dopamine D(3) receptor antagonist offers the potential for an effective antipsychotic therapy, free of the serious side effects of currently available drugs. Using clearance and brain penetration studies as a screen, a series of 1,2,3, 4-tetrahydroisoquinolines, exemplified by 13, was identified with high D(3) affinity and selectivity against the D(2) receptor. Following examination of molecular models, the flexible butyl linker present in 13 was replaced by a more conformationally constrained cyclohexylethyl linker, leading to compounds with improved oral bioavailability and selectivity over other receptors. Subsequent optimization of this new series to improve the cytochrome P450 inhibitory profile and CNS penetration gave trans-N-[4-[2-(6-cyano-1, 2,3, 4 tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-4-quinolinecarbo xamide (24, SB 277011). This compound is a potent and selective dopamine D(3) receptor antagonist with high oral bioavailability and brain penetration in the rat and represents an excellent new chemical tool for the investigation of the role of the dopamine D(3) receptor in the CNS. PMID- 10794705 TI - Structure-based design, synthesis, and biological evaluation of novel pyrrolyl aryl sulfones: HIV-1 non-nucleoside reverse transcriptase inhibitors active at nanomolar concentrations. AB - Pyrrolyl aryl sulfones (PASs) have been recently reported as a new class of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) inhibitors acting at the non-nucleoside binding site of this enzyme (Artico, M.; et al. J. Med. Chem. 1996, 39, 522-530). Compound 3, the most potent inhibitor within the series (EC(50) = 0.14 microM, IC(50) = 0.4 microM, and SI > 1429), was then selected as a lead compound for a synthetic project based on molecular modeling studies. Using the three-dimensional structure of RT cocrystallized with the alpha-APA derivative R95845, we derived a model of the RT/3 complex by taking into account previously developed structure-activity relationships. Inspection of this model and docking calculations on virtual compounds prompted the design of novel PAS derivatives and related analogues. Our computational approach proved to be effective in making qualitative predictions, that is in discriminating active versus inactive compounds. Among the compounds synthesized and tested, 20 was the most active one, with EC(50) = 0.045 microM, IC(50) = 0.05 microM, and SI = 5333. Compared with the lead 3, these values represent a 3- and 8-fold improvement in the cell-based and enzyme assays, respectively, together with the highest selectivity achieved so far in the PAS series. PMID- 10794706 TI - Subtype-selective N-methyl-D-aspartate receptor antagonists: benzimidazalone and hydantoin as phenol replacements. AB - Previous work in our laboratories investigating compounds with structural similarity to ifenprodil (5) and 6 (CP101,606) resulted in compound 7 as a potent and selective antagonist of the NR1/2B subtype of the NMDA receptors. Replacement of the phenol group of 7 with a benzimidazalone group tethered by a three-carbon chain to 4-benzylpiperidine resulted in a slightly less active, but selective, compound. Lengthening the carbon tether resulted in a decrease in NR1/2B potency. Replacement of the phenol ring with a hydantoin resulted in weak antagonists. Compound 11a was one of the most potent NR1/2B antagonists from this study. Compound 11a also potentiated the effects of L-DOPA in a rat model of Parkinson's disease (the 6-hydroxydopamine-lesioned rat), dosed at 30 mg/kg orally. PMID- 10794708 TI - Intracellular green fluorescent protein-polyalanine aggregates are associated with cell death. AB - Eight diseases, exemplified by Huntington's disease and spinocerebellar ataxia type 1, are caused by CAG-repeat expansion mutations. The CAG repeats are translated into expanded polyglutamine tracts, which are associated with deleterious novel functions. While these diseases are characterized by intraneuronal aggregate formation, it is unclear whether the aggregates cause disease. We have addressed this debate by generating intracellular aggregates with green fluorescent protein (GFP) fused to 19-37 alanines. No aggregates were seen in cells expressing native GFP or GFP fused to seven alanines. Aggregate containing cells expressing GFP fused to 19-37 polyalanines show high rates of nuclear fragmentation compared with cells expressing the same constructs without aggregates, or cells expressing GFP fused to seven alanines. This suggests an association between aggregate formation and cell death. PMID- 10794707 TI - Role of N-oligosaccharide endoplasmic reticulum processing reactions in glycoprotein folding and degradation. AB - The endoplasmic reticulum (ER) is the subcellular site where proteins following the secretory pathway acquire their proper tertiary and, in certain cases, quaternary structures. Species that are not yet properly folded are prevented from exit to the Golgi apparatus and, if permanently misfolded, are transported to the cytosol, where they are degraded in the proteasomes. This review deals with a mechanism, applicable to proteins that are N-glycosylated in the ER, by which the quality control of folding is performed. Protein-linked monoglucosylated glycans, formed by glucosidase I- and glucosidase II-dependent partial deglucosylation of the oligosaccharides transferred from dolichol diphosphate derivatives in N-glycosylation (Glc(3)Man(9)GlcNAc(2)), mediate glycoprotein recognition by two ER-resident lectins, membrane-bound calnexin (CNX) and its soluble homologue, calreticulin (CRT). A still not yet fully confirmed interaction between the lectins and the protein moieties of folding glycoproteins may occur after lectin recognition of monoglucosylated structures. Further deglucosylation of glycans by glucosidase II, and perhaps also by a change in CNX/CRT and/or in the substrate glycoprotein conformation, liberates the glycoproteins from their CNX/CRT anchors. Glycans may be then reglucosylated by the UDP-Glc:glycoprotein glucosyltransferase (GT), and thus be recognized again by CNX/CRT, but only when linked to not yet properly folded protein moieties, as this enzyme behaves as a sensor of glycoprotein conformation. Deglucosylation/reglucosylation cycles catalysed by the opposing activities of glucosidase II and GT only stop when proper folding is achieved. The interaction between CNX/CRT and a monoglucosylated glycan is one of the alternative mechanisms by which cells retain not yet properly folded glycoproteins in the ER; in addition, it enhances folding efficiency by preventing protein aggregation and thus allowing intervention of classical chaperones and other folding-assisting proteins. There is evidence suggesting that both glycoprotein glucosylation and mannose removal, respectively mediated by GT and ER mannosidase I, might be involved in cell recognition of permanently misfolded glycoproteins bound for proteasome degradation. PMID- 10794709 TI - Cloning and characterization of ADAM28: evidence for autocatalytic pro-domain removal and for cell surface localization of mature ADAM28. AB - The metalloprotease disintegrins are a family of membrane-anchored glycoproteins with diverse functions in fertilization, myoblast fusion, neurogenesis and protein ectodomain shedding. Here we report a cDNA sequence, encoding a metalloprotease disintegrin, termed ADAM28 ('a disintegrin and metalloprotease 28'), which was cloned from mouse lung. From protein sequence comparisons, ADAM28 is more closely related to snake venom metalloproteases (SVMPs) than to other ADAMs, and hence may cleave similar substrates to SVMPs, perhaps including components of the extracellular matrix. Northern blot analysis of selected mouse tissues revealed that ADAM28 is expressed highly and in alternatively spliced forms in the epididymis, suggesting a possible role in sperm maturation, and at lower levels in lung. The intracellular maturation of ADAM28 expressed in COS-7 cells resembles that of other ADAMs, in that ADAM28 is made as a precursor and processed to a mature form in a late Golgi compartment of the secretory pathway. Most or all of the mature, and thus presumably catalytically active, form of ADAM28 in COS-7 cells is accessible to cell surface trypsinization, suggesting that ADAM28 functions mainly on the cell surface. A mutation converting the catalytic-site glutamate residue into alanine abolishes pro-domain removal, even though this mutant form of ADAM28 can be transported to the cell surface in a manner similar to the wild-type protein. This suggests that pro-domain removal and maturation of ADAM28 may be, at least in part, autocatalytic. This is in contrast with several other ADAMs, for which furin-like proprotein convertases are involved in pro-domain removal, and in which a glutamate-to-alanine mutation in the catalytic site does not alter pro-domain removal. PMID- 10794711 TI - Roles of individual EF-hands in the activation of m-calpain by calcium. AB - m-Calpain is a heterodimeric, cytosolic, thiol protease, which is activated by Ca(2+)-binding to EF-hands in the C-terminal domains of both subunits. There are four potential Ca(2+)-binding EF-hands in each subunit, but their relative affinities for Ca(2+) are not known. In the present study mutations were made in both subunits to reduce the Ca(2+)-binding affinity at one or more EF-hands in one or both subunits. X-ray crystallography of some of the mutated small subunits showed that Ca(2+) did not bind to the mutated EF-hands, but that its binding at other sites was not affected. The structures of the mutant small subunits in the presence of Ca(2+) were otherwise identical to that of the Ca(2+)-bound wild-type small subunit. In the whole enzyme the wild-type macroscopic Ca(2+) requirement (K(d)) was approx. 350 microM. The mutations did not affect the maximum specific activity of the enzyme, but caused increases in K(d), which were characteristic of each site. All the EF-hands could be mutated in various combinations without loss of activity, but preservation of at least one wild-type EF-hand 3 sequence was required to maintain K(d) values lower than 1 mM. The results suggest that all the EF-hands can contribute co-operatively to calpain activation, but that EF hand 3, in both subunits, has the highest intrinsic affinity for Ca(2+) and provides the major driving force for conformational change. PMID- 10794710 TI - Expression of human hyaluronan synthases in response to external stimuli. AB - In the present study we have investigated the expression of mRNAs for hyaluronan synthase isoforms (HAS1, HAS2 and HAS3) in different cells in response to various stimuli. Human mesothelial cells, which synthesize large amounts of hyaluronan, express mRNAs encoding all three HAS isoforms, whereas their transformed counterparts, mesothelioma cells, which produce only minute amounts of hyaluronan, express only HAS3 mRNA. Human lung fibroblasts and the glioma cell line U-118 MG express only the HAS2 and HAS3 genes. The expression of the transcripts was higher in subconfluent than in confluent cultures and was well correlated with the production of hyaluronan by the cells. Stimulation of mesothelial cells with platelet-derived growth factor-BB induced an up-regulation of mRNA for HAS2 to a maximum after 6 h of stimulation; HAS1 and HAS3 genes were only induced slightly. Transforming growth factor-beta1 reduced HAS2 mRNA slightly, and hydrocortisone reduced it strongly, within 6 h of stimulation in mesothelial cell cultures but did not significantly affect the expression of mRNAs for HAS1 and HAS3. Induction of HAS1 and HAS2 protein levels in response to the stimuli above correlated with HAS transcript levels. Thus the expression of the three HAS isoforms is more prominent in growing cells than in resting cells and is differentially regulated by various stimuli suggesting distinct functional roles of the three proteins. PMID- 10794712 TI - Identification of an involucrin promoter transcriptional response element with activity restricted to keratinocytes. AB - The involucrin proximal promoter was examined for response elements that confer cell-type specificity. Using a segment spanning positions -157 to +41, three possible response elements were identified by their protein-binding activity using DNase I footprinting. From distal to proximal, they were: an activator protein-1 (AP-1) site (previously identified) overlapping an Ets-like site; a second Ets-like site located 13 bp more proximally; and an extended region designated footprinted site A (FPA). Mutation of the distal Ets-like site had essentially no effect on the transcriptional activity in transfections, while mutation of the proximal site reduced the activity by half. FPA was shown by electrophoretic mobility-shift assay (EMSA) to be comprised of two separable binding sites, FPA1 (distal) and FPA2 (proximal). While mutation of FPA2 had only a modest effect on transcriptional activity in transient transfections, mutation of FPA1 reduced transcriptional activity to approx. 20% of that obtained with the intact promoter. Additional mutations of FPA1 indicated that the active region comprises positions -85 to -73 (GTGGTGAAACCTGT). The molecular masses of the major proteins binding to this site were shown by UV cross-linking to be approx. 40 and 50 kDa, while minor bands were observed at 80 and 110 kDa. Since the involucrin promoter exhibits much higher transcriptional activity in keratinocytes than in other cell types in transfection assays (indicating that cell type specificity of expression is retained), the comparative influence of FPA1 was examined. While mutation of the AP-1 site affected transcriptional activity similarly in all cell lines tested, mutation of FPA1 decreased activity substantially in keratinocytes, but not in NIH-3T3 and HeLa cells, evidence for a contribution to cell-type specificity of expression. Furthermore, a correlation between the sensitivity to FPA1 mutation and amount of involucrin expression in different keratinocyte cell lines was evident. EMSA showed that NIH-3T3 and HeLa cells lacked the same FPA1 DNA-protein complex as keratinocytes. However, the amount of complex formed with nuclear extracts from several keratinocyte lines did not correlate well with the level of involucrin expression. Other factors, such as differences in post-translational modification or co-activators, must account for varied transcriptional response mediated by this site among keratinocyte lines. PMID- 10794713 TI - Identification and expression analysis of leptin-regulated immediate early response and late target genes. AB - Using PC12 cells as an in vitro model system, we have identified a series of transcripts induced through activation of the leptin receptor. On the basis of kinetic studies, two distinct gene sets could be discerned: signal transducer and activator of transciption-3 (STAT-3), suppressor of cytokine signalling-3 (SOCS 3), MT-II (metallothionein-II), the serine/threonine kinase fibroblast-growth factor-inducible kinase (Fnk) and modulator recognition factor (MRF-1), which are immediate early response genes, and pancreatitis-associated protein I (PAP I), squalene epoxidase, uridine diphosphate glucuronosyltransferase and annexin VIII, which are late induced target genes. At late time points a strong co-stimulation with beta-nerve growth factor or with the adenylate cyclase activator forskolin was observed. To assess the validity of the PC12-cell model system, we examined the effect of leptin administration on the gene transcription of STAT-3, MT-II, Fnk and PAP I in vivo. Leptin treatment of leptin-deficient ob/ob mice increased the STAT-3, SOCS-3, MT-II and Fnk mRNA, and MT-I protein levels in liver, whereas, in jejunum, expression of PAP I mRNA was down-regulated. Furthermore, administration of leptin to starved wild-type mice enhanced the expression of MT II and Fnk mRNA in liver, but decreased MT-II and PAP I mRNA expression in jejunum. These findings may help to explain the obese phenotype observed in some colonies of MT-I- and MT-II-null mice and/or the observation that leptin protects against tumour-necrosis-factor toxicity in vivo. PMID- 10794714 TI - Exon-skipping in BCR/ABL is induced by ABL exon 2. AB - The BCR/ABL fusion gene is pathognomonic for chronic myelogenous leukaemia (CML). We have previously reported alternative splicing of BCR/ABL, as indicated by the detection of both p190- and p210-encoding transcripts, in about 60% of CML patient samples. These exon-skipping events involved the joining of ABL exon 2 to variable upstream BCR exons. Similarly, ABL exon 2 is alternatively spliced to either of two upstream ABL exons (1a or 1b) in c-ABL. We have constructed BCR and BCR/ABL minigenes to study this phenomenon in more detail. These constructs were transfected into various cell types and splicing was assessed by reverse transcriptase PCR. Whereas the basic BCR minigene expressed exon-inclusive transcripts only, insertion of genomic DNA spanning ABL exon 2 induced exon skipping but only when expressed in the CML cell lines K562 and EM3. In this study we localized the required sequence element to ABL exon 2 itself. These results mimic the splicing phenotype displayed by most CML patients. We propose a model where a trans-factor present in some CML cells interacts with ABL exon 2 pre-mRNA to promote skipping of upstream BCR exons. PMID- 10794715 TI - Sphingosine 1-phosphate stimulates proliferation and migration of human endothelial cells possibly through the lipid receptors, Edg-1 and Edg-3. AB - Sphingosine 1-phosphate (S1P) stimulates thymidine incorporation (DNA synthesis), cell growth and cell migration in human aortic endothelial cells (HAECs). The extent of the S1P-induced responses are comparable to those stimulated by vascular endothelial growth factor, one of the most potent stimulators of angiogenesis. These responses to S1P were mimicked by dihydrosphingosine 1 phosphate, an S1P receptor agonist, and inhibited by pertussis toxin (PTX), an inactivator of G(i)/G(o)-proteins. S1P also induced activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (p38 MAP kinase). The activation of these enzymes was inhibited again by PTX and also by suramin, a non-selective receptor antagonist. S1P-induced DNA synthesis and ERK activation were inhibited by PD98059, an ERK kinase inhibitor, but not by SB203580, a p38 MAP kinase inhibitor. In contrast, cell migration and p38 MAP kinase activation, in response to S1P, were inhibited by SB203580 but not by PD98059. In HAECs, high-affinity S1P binding activity and expression of Edg-1 and Edg-3 mRNA were detected. These results suggest that S1P might be a novel angiogenesis factor and that the lipid-induced proliferation and migration of endothelial cells are possibly mediated through cell-surface S1P receptors, Edg-1 and Edg-3, which are linked to signalling pathways. PMID- 10794716 TI - Mutational analysis of Lys65 of HIV-1 reverse transcriptase. AB - Amino acid Lys(65) is part of the highly flexible beta3-beta4 loop in the fingers domain of the 66 kDa subunit of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT). Recent crystal data show that the epsilon-amino group of Lys(65) interacts with the gamma-phosphate of the bound deoxynucleoside triphosphate ('dNTP') substrate [Huang, Chopra, Verdine and Harrison (1998) Science 282, 1669-1675]. In order to biochemically define the function of RT Lys(65), we have used site-specific mutagenesis to generate RT with a variety of substitutions at this position, including K65E, K65Q, K65A and K65R. Kinetic analyses demonstrate that if Lys(65) in RT is substituted with an amino acid other than arginine the enzyme exhibits dramatic decreases in the binding affinity (K(m)) for all dNTP substrates, in RT catalytic efficiency (k(cat)/K(m)) and in the mutant enzyme's ability to carry out pyrophosphorolysis, the reverse reaction of DNA synthesis. The pH optimum for the DNA polymerase activity of K65E RT was 6.5, compared to 7.5 for the wild-type enzyme, and 8.0 for the K65R, K65A and K65Q mutants. Molecular modelling studies show that mutations of Lys(65) do not affect the geometry of the loop's alpha-carbon backbone, but rather lead to changes in positioning of the side chains of residues Lys(70) and Arg(72). In particular, Glu in K65E can form a salt bridge with Arg(72), leading to the diminution of the latter residue's interaction with the alpha-phosphate of the dNTP residue. This alteration in dNTP-binding may explain the large pH-dependent changes in both dNTP-binding and catalytic efficiency noted with the enzyme. Furthermore, the K65A, K65Q and K65E mutant enzymes are 100-fold less sensitive to all dideoxynucleoside triphosphate ('ddNTP') inhibitors, whereas the K65R mutation results in a selective 10-fold decrease in binding of ddCTP and ddATP only. This implies that mutations at position 65 in HIV-1 RT influence the nucleotide-binding specificity of the enzyme. PMID- 10794717 TI - Chronic insulin effects on insulin signalling and GLUT4 endocytosis are reversed by metformin. AB - Decreases in insulin-responsive glucose transport and associated levels of cell surface GLUT4 occur in rat adipocytes maintained in culture for 20 h under hyperinsulinaemic and hyperglycaemic conditions. We have investigated whether this defect is due to reduced signalling from the insulin receptor, GLUT4 expression or impaired GLUT4 trafficking. The effects of chronic insulin treatment on glucose transport and GLUT4 trafficking were ameliorated by inclusion of metformin in the culture medium. In comparison with the ic insulin treatment attenuated changes in signalling processes leading to glucose transport. These included insulin receptor tyrosine phosphorylation, phosphoinositide 3-kinase activity and Akt activity, which were all reduced by 60 70%. Inclusion of metformin in the culture medium prevented the effects of the chronic insulin treatment on these signalling processes. In comparison with cells maintained in culture without insulin, the total expression of GLUT4 protein was not significantly altered by chronic insulin treatment, although the level of GLUT1 expression was increased. Trafficking rate constants for wortmannin-induced cell-surface loss of GLUT4 and GLUT1 were assessed by 2-N-4-(1-azi-2, 2,2 trifluoroethyl)benzoyl-1,3-bis(D-mannose-4-yloxy)-2-propyla min e (ATB-BMPA) photolabelling. In comparison with cells acutely treated with insulin, chronic insulin treatment resulted in a doubling of the rate constants for GLUT4 endocytosis. These results suggest that the GLUT4 endocytosis process is very sensitive to the perturbations in signalling that occur under hyperinsulinaemic and hyperglycaemic conditions, and that the resulting elevation of endocytosis accounts for the reduced levels of net GLUT4 translocation observed. PMID- 10794718 TI - JNK (c-Jun N-terminal kinase) and p38 activation in receptor-mediated and chemically-induced apoptosis of T-cells: differential requirements for caspase activation. AB - Activation of the stress-activated mitogen-activated protein kinases (MAP kinases), c-Jun N-terminal kinase (JNK) and p38, is necessary for the induction of apoptosis in neuronal cells; however, in other cell types their involvement may be stimulus-dependent. In the present study we investigate the activation of JNK and p38 in a single non-neuronal cell type, undergoing receptor-mediated (tumour necrosis factor-related apoptosis-inducing ligand and CD95) or chemically induced (lactacystin) apoptosis. In Jurkat T-cells, receptor-mediated and chemically-induced apoptosis resulted in a time-dependent activation of the initiator caspases-8 and -9, respectively. Both types of stimuli resulted in a significant activation of JNK and p38, which closely paralleled the time dependent induction of apoptosis. The caspase inhibitor, benzyloxycarbonyl-Val Ala-Asp-(OMe) fluoromethyl ketone (z-VAD.FMK) inhibited receptor-mediated apoptosis and suppressed JNK and p38 activation. In contrast, inhibition of lactacystin-induced apoptosis with z-VAD.FMK, as assessed by phosphatidylserine exposure and poly(ADP-ribose) polymerase cleavage, did not inhibit activation of JNK or p38, demonstrating that during chemically-induced apoptosis, activation of JNK and p38 is independent of effector caspases. The role of p38 in apoptosis was assessed using the specific p38 inhibitor, SB203580. No effect on the induction of apoptosis or caspase activation was observed, although activation of mitogen activated protein kinase-activated protein kinase-2 (MAPKAPK-2), an immediate downstream target of p38, was inhibited. Therefore neither p38 activation nor activation of MAPKAPK-2 is critical for induction of either receptor- or chemically-induced apoptosis. Thus, within a single cell type, (1) the mechanism of p38 and JNK activation during apoptosis is stimulus-dependent and (2) activation of the p38 pathway is not required for caspase activation or apoptosis, assessed by phosphatidylserine exposure, but may still be required to elicit other features of the apoptotic phenotype. PMID- 10794719 TI - Membrane-induced conformational change in human apolipoprotein H. AB - The interaction of apolipoprotein H (Apo H) with lipid membrane has been considered to be a basic mechanism for the biological function of the protein. Previous reports have demonstrated that Apo H can interact only with membranes containing anionic phospholipids. Here we study the membrane-induced conformational change of Apo H by CD spectroscopy with two different model systems: anionic-phospholipid-containing liposomes [such as 1, 2-dimyristoyl-sn glycero-3-phosphoglycerol (DMPG) and cardiolipin], and the water/methanol mixtures at moderately low pH, which mimic the micro-physicochemical environment near the membrane surface. It is found that Apo H undergoes a remarkable conformational change on interaction with liposomes containing anionic phospholipid. To interact with liposomes containing DMPG, there is a 6.8% increase in alpha-helix in the secondary structures; in liposomes containing cardiolipin, however, there is a 12.6% increase in alpha-helix and a 9% decrease in beta-sheet. The similar conformation change in Apo H can be induced by treatment with an appropriate mixture of water/methanol. The results indicate that the association of Apo H with membrane is correlated with a certain conformational change in the secondary structure of the protein. PMID- 10794720 TI - pp60c-src associates with the SH2-containing inositol-5-phosphatase SHIP1 and is involved in its tyrosine phosphorylation downstream of alphaIIbbeta3 integrin in human platelets. AB - SH2-containing inositol-5-phosphatase 1 (SHIP1) was originally identified as a 145 kDa protein that became tyrosine-phosphorylated in response to multiple cytokines. It is now well established that SHIP1 is specifically expressed in haemopoietic cells and is important as a negative regulator of signalling. We found recently that SHIP1 was present in human blood platelets as an Ins(1,3,4, 5)P(4)-phosphatase and a PtdIns(3,4,5)P(3)-5-phosphatase that became tyrosine phosphorylated and was relocated to the cytoskeleton in an integrin-dependent manner. Here we report biochemical and pharmacological evidence that the tyrosine kinase pp60(c-src) is constitutively associated with SHIP1 and is involved in its tyrosine phosphorylation downstream of integrin engagement in thrombin-activated human platelets. The use of cytochalasin D allowed us to demonstrate that the actin cytoskeleton reorganization induced on thrombin stimulation was not required for its integrin-mediated phosphorylation. Moreover, the integrin dependent relocation of SHIP1 to the cytoskeleton did not require its tyrosine phosphorylation. These results suggest that SHIP1 is first recruited to the integrin-linked signalling complexes and then becomes tyrosine-phosphorylated through a Src-kinase-dependent mechanism but independently of the actin cytoskeleton reorganization. PMID- 10794721 TI - Lactoferrin binding to the rat asialoglycoprotein receptor requires the receptor's lectin properties. AB - Lactoferrin binds to rat hepatic lectin 1 (RHL1), the major subunit of the asialoglycoprotein (ASGP) receptor, with high affinity, by a galactose independent mechanism. To better understand the molecular basis of this novel interaction, we compared the binding of lactoferrin and asialo-orosomucoid (ASOR) to isolated rat hepatocytes and to purified ASGP receptors as a function of pH, Ca(2+) and receptor acylation. Binding of (125)I-lactoferrin and (125)I-ASOR to isolated rat hepatocytes at 4 degrees C decreased sharply at pH<6, following similar titration curves. Binding of (125)I-lactoferrin and (125)I-ASOR to hepatocytes was Ca(2+)-dependent. Binding increased progressively at > or =300 microM CaCl(2), in the presence of 1 mM EDTA. Monensin treatment of hepatocytes, which causes hepatocytes to accumulate inactive ASGP receptors, reduced surface binding of (125)I-lactoferrin and (125)I-ASOR by 46 and 49%, respectively, with only a 16% loss of immunodetectable receptor protein from the cell surface. Finally, deacylation of purified ASGP receptors in vitro with 1 M hydroxylamine abolished receptor lectin activity as reflected by the loss of (125)I-ASOR binding as well as the complete loss of specific (125)I-lactoferrin binding. Treatment with 1 M Tris had no effect on binding of either ligand. We conclude from these data that galactose-independent lactoferrin binding to the ASGP receptor requires the receptor's carbohydrate-recognition domain to be in an active configuration. An active configuration is promoted by neutral pH and Ca(2+), and also requires the receptor subunits to be acylated. PMID- 10794723 TI - Net release of individual fatty acids from white adipose tissue during lipolysis in vitro: evidence for selective fatty acid re-uptake. AB - During lipolysis, adipose tissue triacylglycerols (TAG) undergo concurrent breakdown and synthesis because some of the newly hydrolysed and released non esterified ('free') fatty acids (NEFA) can subsequently be taken up and re esterified. The present study examines whether and how the release of individual fatty acids is affected by the re-uptake of some of the newly hydrolysed fatty acids in vitro during lipolysis. To alter fatty acid release and re-uptake, adipose tissue fragments and isolated adipocytes from rats were incubated under various conditions, i.e. several cell concentrations or adipose fragment quantities, with or without glucose. In the various conditions tested, the NEFA/glycerol molar ratio ranged from 1.5 to 2.9. Whatever the incubation conditions, including those resulting in very low, medium or high fatty acid re uptake (as assessed by the NEFA/glycerol ratio), the percentage weight of fatty acids in NEFA was significantly different from that in TAG for 20-24 of the 35 fatty acids that were considered. Thus the greater the fatty acid re-uptake, the higher the proportion of polyunsaturated fatty acids and the lower the proportion of long-chain saturated and monounsaturated fatty acids in NEFA. Moreover, the relative mobilization (%NEFA/%TAG) of the least readily mobilized fatty acid (C(22:1,n-11)) was 6.2-fold lower than that of the most readily mobilized fatty acid (C(20:5,n-3)) under conditions of very low fatty acid re-uptake, and 14.8 fold lower under conditions of high fatty acid re-uptake, indicating a widening of the range of relative mobilizations. We conclude that the composition of the NEFA pool is affected by the rate of fatty acid re-uptake. This provides strong evidence for the selective re-uptake of adipose tissue fatty acids during lipolysis. PMID- 10794722 TI - Autophosphorylation of Tyr397 and its phosphorylation by Src-family kinases are altered in focal-adhesion-kinase neuronal isoforms. AB - In brain, focal adhesion kinase (FAK) is regulated by neurotransmitters and has a higher molecular mass than in other tissues, due to alternative splicing. Two exons code for additional peptides of six and seven residues ('boxes' 6 and 7), located on either side of Tyr(397), which increase its autophosphorylation. Using in situ hybridization and a monoclonal antibody (Mab77) which does not recognize FAK containing box 7, we show that, although mRNAs coding for boxes 6 and 7 have different patterns of expression in brain, FAK+6,7 is the main isoform in forebrain neurons. The various FAK isoforms fused to green fluorescent protein were all targeted to focal adhesions in non-neuronal cells. Phosphorylation-state specific antibodies were used to study in detail the phosphorylation of Tyr(397), a critical residue for the activation and function of FAK. The presence of boxes 6 and 7 increased autophosphorylation of Tyr(397) independently and additively, whereas they had a weak effect on FAK kinase activity towards poly(Glu,Tyr). Src family kinases were also able to phosphorylate Tyr(397) in cells, but this phosphorylation was decreased in the presence of box 6 or 7, and abolished in the presence of both. Thus the additional exons characteristic of neuronal isoforms of FAK do not alter its targeting, but change dramatically the phosphorylation of Tyr(397). They increase its autophosphorylation in vitro and in transfected COS-7 cells, whereas they prevent its phosphorylation when co-transfected with Src family kinases. PMID- 10794724 TI - Fractionation and characterization of oligomeric, protofibrillar and fibrillar forms of beta-amyloid peptide. AB - The beta-amyloid (Abeta) peptide, a major component of senile plaques in Alzheimer's disease brain, has been shown previously to undergo a process of polymerization to produce neurotoxic forms of amyloid. Recent literature has attempted to define precisely the form of Abeta responsible for its neurodegenerative properties. In the present study we describe a novel density gradient centrifugation method for the isolation and characterization of structurally distinct polymerized forms of Abeta peptide. Fractions containing protofibrils, fibrils, sheet structures and low molecular mass oligomers were prepared. The fractionated forms of Abeta were characterized structurally by transmission electron microscopy. The effects on cell viability of these fractions was determined in the B12 neuronal cell line and hippocampal neurons. Marked effects on cell viability in the cells were found to correspond to the presence of protofibrillar and fibrillar structures, but not to monomeric peptide or sheet-like structures of polymerized Abeta. Biological activity correlated with a positive reaction in an immunoassay that specifically detects protofibrillar and fibrillar Abeta; those fractions that were immunoassay negative had no effect on cell viability. These data suggest that the effect of Abeta on cell viability is not confined to a single conformational form but that both fibrillar and protofibrillar species have the potential to be active in this assay. PMID- 10794725 TI - Inhibition by long-chain acyl-CoAs of glucose 6-phosphate metabolism in plastids isolated from developing embryos of oilseed rape (Brassica napus L.). AB - The effects of long-chain acyl-CoA (lcACoA) esters (both added exogenously and synthesized de novo) and acyl-CoA binding protein (ACBP) on plastidial glucose 6 phosphate (Glc6P) and pyruvate metabolism were examined using isolated plastids. The binding of lcACoA esters by ACBP stimulated the utilization of Glc6P for fatty acid synthesis, starch synthesis and reductant supply via the oxidative pentose phosphate (OPP) pathway. Stimulation occurred at low (1-10 microM) concentrations of ACBP. Pyruvate-dependent fatty acid synthesis was not directly affected by ACBP. However, addition of ACBP did increase the Glc6P-dependent stimulation of pyruvate utilization mediated through the OPP pathway. On the basis of these experiments, we conclude that lcACoA esters may inhibit Glc6P uptake into plastids, and that this inhibition is relieved by ACBP. We also suggest that utilization of other substrates for fatty acid synthesis may be affected by lcACoA/ACBP via their effects on the OPP pathway. PMID- 10794727 TI - Identification of amino acids imparting acceptor substrate selectivity to human arylamine acetyltransferases NAT1 and NAT2. AB - The human arylamine N-acetyltransferases NAT1 and NAT2 catalyse the acetyl-CoA dependent N- and O-acetylation of primary arylamine and hydrazine xenobiotics and their N-hydroxylated metabolites. We previously used a panel of recombinant NAT1/NAT2 chimaeric proteins to identify linear amino acid segments that have roles in imparting the distinct catalytic specificities to these proteins [Dupret, Goodfellow, Janezic and Grant (1994) J. Biol. Chem. 269, 26830-26835]. These studies indicated that a conserved central region (residues 112-210) distinct from that containing the active-site cysteine residue Cys(68) was important in determining NAT substrate selectivity. In the present study we have refined our analysis through further chimaera generation of this conserved region and by subsequent site-directed mutagenesis of individual amino acids. Enzyme kinetic analysis of these mutant proteins with the NAT1-selective and NAT2 selective substrates p-aminosalicylic acid (PAS) and sulphamethazine (SMZ) respectively suggests that residues 125, 127 and 129 are important determinants of NAT1-type and NAT2-type substrate selectivity. Modification of Arg(127) had the greatest effect on specificity for PAS, whereas changing Phe(125) had the greatest effect on specificity for SMZ. Selected NAT mutants exhibited K(m) values for acetyl-CoA that were comparable with those of the wild-type NATs, implying that the mutations affected acceptor substrate specificity rather than cofactor binding affinity. Taken together with previous observations, these results suggest that residues 125, 127 and 129 might contribute to the formation of the active-site pocket surrounding Cys(68) and function as important determinants of NAT substrate selectivity. PMID- 10794726 TI - Heterogeneous nuclear ribonucleoprotein D0 contains transactivator and DNA binding domains. AB - Heterogeneous nuclear ribonucleoprotein D0 (hnRNP D0) is an abundant, ubiquitous protein that binds RNA and DNA sequences specifically, and has been implicated in the transcriptional regulation of the human complement receptor 2 gene. We found that in vivo expression of hnRNP D0-GAL4 fusion proteins increased the transcriptional activity of a GAL4-driven reporter gene, providing direct proof that hnRNP D0 possesses a transactivator domain. We found, using truncated hnRNP D0 proteins fused to GAL4, that 29 amino acids in the N-terminal region are critical for transactivation. We established, using a series of recombinant truncated hnRNP D0 proteins, that the tandem RNA-binding domains alone were not able to bind double-stranded DNA. Nevertheless, 24 additional amino acids of the C-terminus imparted sequence-specific DNA binding. Experiments using peptide specific antisera supported the importance of the 24-amino-acid region in DNA binding, and suggested the involvement of the 19-amino-acid alternative insert which is present in isoforms B and D. The N-terminus had an inhibitory effect on binding of hnRNP D0 to single-stranded, but not to double-stranded, DNA. Although both recombinant hnRNP D0B and D0D bound DNA, only the B isoform recognized DNA in vivo. We propose that the B isoform of hnRNP D0 functions in the nucleus as a DNA-binding transactivator and has distinct transactivator and DNA-binding domains. PMID- 10794728 TI - Designing transthyretin mutants affecting tetrameric structure: implications in amyloidogenicity. AB - The molecular mechanisms that convert soluble transthyretin (TTR) tetramers into insoluble amyloid fibrils are still unknown; dissociation of the TTR tetramer is a pre-requisite for amyloid formation in vitro and involvement of monomers and/or dimers in fibril formation has been suggested by structural studies. We have designed four mutated proteins with the purpose of stabilizing [Ser(117)-->Cys (S117C) and Glu(92)-->Cys (E92C)] or destabilizing [Asp(18)-->Asn (D18N) and Leu(110)-->Ala (D110A)] the dimer/tetramer interactions in TTR, aiming at elucidating structural determinants in amyloidogenesis. The resistance of the mutated proteins to dissociation was analysed by HPLC studies of diluted TTR preparations. Both 'stabilized' mutants migrated as tetramers and, upon dilution, no other TTR species was observed, confirming the increased resistance to dissociation. For the 'destabilized' mutants, a mixture of tetrameric and monomeric forms co-existed at low dilution and the latter increased upon 10-fold dilution. Both of the destabilizing mutants formed amyloid in vitro when acidified. This result indicated that both the AB loop of TTR, destabilized in D18N, and the hydrophobic interactions affecting the dimer-dimer interfaces in L110A are implicated in the stability of the tetrameric structure. The stabilized mutants, which were dimeric in nature through disulphide bonding, were unable to polymerize into amyloid, even at pH 3.2. When the amyloid formation assay was repeated in the presence of 2-mercaptoethanol, upon disruption of the S-S bridges of these stable dimers, amyloid fibril formation was observed. This experimental evidence suggests that monomers, rather than dimers, are the repeating structural subunit comprising the amyloid fibrils. PMID- 10794730 TI - Electron-microscopic demonstration of multidrug resistance protein 2 (Mrp2) retrieval from the canalicular membrane in response to hyperosmolarity and lipopolysaccharide. AB - Immunohistochemical studies suggest that canalicular secretion via multidrug resistance protein 2 (Mrp2), a conjugate export pump encoded by the Mrp2 gene, is regulated by rapid transporter retrieval from/insertion into the canalicular membrane. The present study was undertaken in order to investigate this suggestion by means of immunogold electron microscopy. Therefore the effects of lipopolysaccharide (LPS) and osmolarity on Mrp2 localization were studied following immunogold labelling in the perfused rat liver by quantitative electron microscopy and morphometric analyses, and by confocal laser scanning microscopy. Mrp2 activity was assessed in the isolated perfused rat liver by measuring the excretion of dinitrophenyl-S-glutathione as a substrate of Mrp2. Both LPS and hyperosmolarity resulted in a statistically significant decrease in immunogold labelled Mrp2 in the canalicular membrane and canalicular villi, and an increase in labelling in the pericanalicular cytoplasm. Canalicular morphometric parameters were unchanged under these conditions compared with controls. Under hyperosmolar perfusion Mrp2, but not the canalicular protein dipeptidylpeptidase IV, was found inside the cells, as shown by double immunofluorescence and confocal laser scanning microscopy. The findings suggest a selective retrieval of Mrp2 from the canalicular membrane under the influence of hyperosmolarity and LPS, whereas canalicular morphology remains unchanged. PMID- 10794729 TI - Regulation of ATP-induced calcium release in COS-7 cells by calcineurin. AB - Experiments were conducted to examine the role of calcineurin in regulating Ca(2+) fluxes in mammalian cells. In COS-7 cells, increasing concentrations (1-10 microM) of ATP triggered intracellular Ca(2+) release in a dose-dependent manner. Treatment of the cells with calcineurin inhibitors such as cyclosporin A (CsA), deltamethrin and FK506 resulted in an enhancement of ATP-induced intracellular Ca(2+) release. Measurement of calcineurin-specific phosphatase activity in vitro demonstrated a high level of endogenous calcineurin activities in COS-7 cells, which was effectively inhibited by the addition of deltamethrin or CsA. The expression of constitutively active calcineurin (CnADeltaCaMAI) inhibited the ATP induced increase in intracellular Ca(2+) concentration ([Ca(2+)](i)), in both the presence and the absence of extracellular Ca(2+). These results suggest that the constitutively active calcineurin prevented Ca(2+) release from the intracellular stores. In the calcineurin-transfected cells, treatment with CsA restored the calcineurin-mediated inhibition of intracellular Ca(2+) release. Protein kinase C mediated phosphorylation of Ins(1,4,5)P(3) receptor [Ins(1,4,5)P(3)R] was partly inhibited by the extracts prepared from the vector-transfected cells and completely inhibited by those from cells co-transfected with CnADeltaCaMAI and calcineurin B. On the addition of 10 microM CsA, the inhibited phosphorylation of Ins(1,4,5)P(3)R was restored in both the vector-transfected cells and the calcineurin-transfected cells. These results show direct evidence that Ca(2+) release through Ins(1, 4,5)P(3)R in COS-7 cells is regulated by calcineurin mediated dephosphorylation. PMID- 10794731 TI - Cloning of human Ca2+ homoeostasis endoplasmic reticulum protein (CHERP): regulated expression of antisense cDNA depletes CHERP, inhibits intracellular Ca2+ mobilization and decreases cell proliferation. AB - A monoclonal antibody which blocks InsP(3)-induced Ca(2+) release from isolated endoplasmic reticulum was used to isolate a novel 4.0 kb cDNA from a human erythroleukaemia (HEL) cell cDNA expression library. A corresponding mRNA transcript of approx. 4.2 kb was present in all human cell lines and tissues examined, but cardiac and skeletal muscle had an additional transcript of 6.4 kb. The identification in GenBank(R) of homologous expressed sequence tags from many tissues and organisms suggests that the gene is ubiquitously expressed in higher eukaryotes. The gene was mapped to human chromosome 19p13.1. The cDNA predicts a 100 kDa protein, designated Ca(2+) homoeostasis endoplasmic reticulum protein (CHERP), with two putative transmembrane domains, multiple consensus phosphorylation sites, a polyglutamine tract of 12 repeats and regions of imperfect tryptophan and histadine octa- and nona-peptide repeats. In vitro translation of the full-length cDNA produced proteins of M(r) 128000 and 100000, corresponding to protein bands detected by Western blotting of many cell types. CHERP was co-localized in HEL cells with the InsP(3) receptor by two-colour immunofluorescence. Transfection of HEL cells with antisense cDNA led to an 80% decline in CHERP within 5 days of antisense induction, with markedly decreased intracellular Ca(2+) mobilization by thrombin, decreased DNA synthesis and growth arrest, indicating that the protein has an important function in Ca(2+) homoeostasis, growth and proliferation. PMID- 10794732 TI - Structure and function of Humicola insolens family 6 cellulases: structure of the endoglucanase, Cel6B, at 1.6 A resolution. AB - Cellulases are traditionally classified as either endoglucanases or cellobiohydrolases on the basis of their respective catalytic activities on crystalline cellulose, which is generally hydrolysed more efficiently only by the cellobiohydrolases. On the basis of the Trichoderma reesei cellobiohydrolase II structure, it was proposed that the active-site tunnel of cellobiohydrolases permitted the processive hydrolysis of cellulose, whereas the corresponding endoglucanases would display open active-site clefts [Rouvinen, Bergfors, Teeri, Knowles and Jones (1990) Science 249, 380-386]. Glycoside hydrolase family 6 contains both cellobiohydrolases and endoglucanases. The structure of the catalytic core of the family 6 endoglucanase Cel6B from Humicola insolens has been solved by molecular replacement with the known T. reesei cellobiohydrolase II as the search model. Strangely, at the sequence level, this enzyme exhibits the highest sequence similarity to family 6 cellobiohydrolases and displays just one of the loop deletions traditionally associated with endoglucanases in this family. However, this enzyme shows no activity on crystalline substrates but a high activity on soluble substrates, which is typical of an endoglucanase. The three-dimensional structure reveals that the deletion of just a single loop of the active site, coupled with the resultant conformational change in a second 'cellobiohydrolase-specific' loop, peels open the active-site tunnel to reveal a substrate-binding groove. PMID- 10794733 TI - Effects of magnesium and nucleotides on the proton conductance of rat skeletal muscle mitochondria. AB - During oxidative phosphorylation most of the protons pumped out to the cytosol across the mitochondrial inner membrane return to the matrix through the ATP synthase, driving ATP synthesis. However, some of them leak back to the matrix through a proton-conductance pathway in the membrane. When the ATP synthase is inhibited with oligomycin and ATP is not being synthesized, all of the respiration is used to drive the proton leak. We report here that Mg(2+) inhibits the proton conductance in rat skeletal-muscle mitochondria. Addition of Mg(2+) inhibited both oligomycin-inhibited respiration and the proton conductance, while removal of Mg(2+) using EDTA activated these processes. The proton conductance was inhibited by more than 80% as free Mg(2+) was raised from 25 nM to 220 microM. Half-maximal inhibition occurred at about 1 microM free Mg(2+), which is close to the contaminating free Mg(2+) concentration in our incubations in the absence of added magnesium chelators. ATP, GTP, CTP, TTP or UTP at a concentration of 1 mM increased the oligomycin-inhibited respiration rate by about 50%. However, these NTP effects were abolished by addition of 2 mM Mg(2+) and any NTP-stimulated proton conductance was explained completely by chelation of endogenous free Mg(2+). The corresponding nucleoside diphosphates (ADP, GDP, CDP, TDP or UDP) at 1 mM had no effect on oligomycin-inhibited respiration. We conclude that proton conductance in rat skeletal-muscle mitochondria is very sensitive to free Mg(2+) concentration but is insensitive to NTPs or NDPs at 1 mM. PMID- 10794734 TI - Glucokinase and glucokinase regulatory protein: mutual dependence for nuclear localization. AB - Conditional expression of the glucokinase regulatory protein in insulinoma cells, under control of the reverse tetracycline-dependent transactivator, was used to investigate whether expression of this protein de novo would alter the intracellular distribution of glucokinase. The regulatory protein, which was undetectable in the basal state, could be induced by doxycycline to levels comparable to those of liver and was detected mostly in the nucleus. Concomitantly, glucokinase accumulated in the nucleus. Human embryonic kidney cells were transiently transfected to express glucokinase and the regulatory protein, either separately or together. Each protein localized predominantly to the cytoplasm when expressed alone. On co-expression, however, both proteins localized virtually entirely to the nucleus. The enzymic activity of glucokinase was not required for promoting nuclear import of the two proteins, as shown with a glucose-phosphorylation-deficient mutant. Finally, in embryonic kidney cells expressing the regulatory protein alone, treatment with leptomycin B resulted in a partial redistribution of the protein from the cytoplasm to the nucleus, suggesting that this protein can shuttle between the two compartments. PMID- 10794735 TI - Comparative aspects of the diffusion of norfloxacin, cefepime and spermine through the F porin channel of Enterobacter cloacae. AB - In Enterobacteriaceae, the permeability of the outer membrane to hydrophilic antibiotics is associated with the presence of pore-forming proteins. We tested the diffusion of the fluoroquinolone norfloxacin in four Enterobacter cloacae strains: a clinical isolate and three derivatives variously producing or lacking the D and F porins. We analysed the entry of norfloxacin into E. cloacae cells in the presence of either the polyamine spermine or the recently developed cefepime, which are known to penetrate through the Escherichia coli OmpF porin. Uptake of the fluoroquinolone was decreased in both cases; the initial rate of penetration decreased as more spermine blocked the channel. Our results indicate that, like beta-lactam molecules, fluoroquinolones translocate through the outer membrane via the F porin and that cefepime and norfloxacin entries are polyamine sensitive. This suggests that the closure of the F porin channel by polyamines might modulate the susceptibility of E. cloacae to both fluoroquinolone and cephalosporin antibiotics. PMID- 10794736 TI - Novel induction of alpha-lactalbumin-mediated lacdiNAc-R expression in vivo. AB - alpha-Lactalbumin (alpha-LA) is a regulatory protein by which the mammalian beta1,4-galactosyltransferase (beta1,4-galT) is induced to utilize glucose as an acceptor instead of N-acetylglucosamine (GlcNAc) during lactose synthesis in mammary gland. alpha-LA can also modulate beta1,4-galT to utilize UDP-N acetylgalactosamine (UDP-GalNAc) as a donor towards GlcNAc acceptor substrate with high efficiency in vitro [Do, Do and Cummings (1995) J. Biol. Chem. 270, 18447-18451]. In the present study we transfected cDNA encoding bovine alpha-LA into Lec8 cells and examined whether nucleotide sugar switching of UDP-galactose (UDP-Gal) into UDP-GalNAc occurred in vivo and whether the neo-glycosylation of GalNAcbeta1,4GlcNAc-R structure was synthesized in alpha-LA-stable transfectants. Our studies demonstrate that the stable expression of alpha-LA in Lec8 cells induces the formation of GalNAcbeta1,4GlcNAc-R in vivo through the nucleotide sugar switching of beta1,4-galT. PMID- 10794738 TI - Finding a needle in a haystack: detection and quantification of rare mutant alleles are coming of age. PMID- 10794739 TI - Current concepts in biomarker technology for bladder cancers. AB - BACKGROUND: Transitional cell carcinoma of the bladder (TCC) is the second most common malignancy of the urinary tract. More than 70% of treated tumors recur, and 30% of recurrent tumors progress. Currently, pathologic staging and grading are valuable prognostic factors for detecting and monitoring TCC. Urinalysis, cystoscopy, and cytology are either invasive or lack sensitivity and specificity. The availability of a noninvasive, reliable, and simple test would greatly improve the detection and monitoring of patients with TCC. Several biomarkers for bladder cancer have been proposed, but no single marker has emerged as the test of choice. APPROACH: We undertook a comprehensive literature search using Medline to identify all publications from 1980 to 1999. Articles that discussed potential biomarkers for TCC were screened. Only compounds that demonstrated high sensitivity or specificity, significant correlation with TCC diagnosis and staging, and extensive investigation were included in this review. CONTENT: Potential biomarkers of disease progression and prognosis include nuclear matrix protein, fibrin/fibrinogen product, bladder tumor antigen, blood group-related antigens, tumor-associated antigens, proliferating antigens, oncogenes, growth factors, cell adhesion molecules, and cell cycle regulatory proteins. The properties of the biomarkers and the methods for detecting or quantifying them are presented. Their sensitivities and specificities for detecting and monitoring disease were 54-100% and 61-97%, respectively, compared with 20-40% and 90% for urinalysis and cytology. SUMMARY: Although urine cytology and cystoscopy are still the standard of practice, many candidate biomarkers for TCC are emerging and being adopted into clinical practice. Further research and better understanding of the biology of bladder cancer, improved diagnostic techniques, and standardized interpretation are essential steps to develop reliable biomarkers. It is possible that using the current biomarkers as an adjuvant modality will improve our ability to diagnose and monitor bladder cancer. PMID- 10794737 TI - The repression of nuclear factor I/CCAAT transcription factor (NFI/CTF) transactivating domain by oxidative stress is mediated by a critical cysteine (Cys-427). AB - The activity of the nuclear factor I/CCAAT transcription factor (NFI/CTF) is negatively regulated by oxidative stress. The addition of relatively high (millimolar) H(2)O(2) concentrations inactivates cellular NFI DNA-binding activity whereas lower concentrations can repress NFI/CTF transactivating function. We have investigated the mechanism of this regulation using Gal4 fusion proteins and transfection assays. We show that micromolar H(2)O(2) concentrations repress the transactivating domain of NFI/CTF in a dose-dependent manner and are less or not active on other transcription factors' transactivating domains. Studies using deletions and point mutations pointed to the critical role of Cys 427. Indeed, when this cysteine is mutated into a serine, the repression by H(2)O(2) is totally blunted. Mutation of other cysteine, serine and tyrosine residues within the transactivating domain had no clear effect on the repression by H(2)O(2). Finally, treatment of cells with the thiol-alkylating reagent N ethylmaleimide leads to a decrease in the transactivating function, which is dependent on Cys-427. This study shows that transactivating domains of transcription factors can constitute very sensitive targets of oxidative stress and highlights the critical role of these domains. PMID- 10794740 TI - Analysis of the chimeric CYP21P/CYP21 gene in steroid 21-hydroxylase deficiency. AB - BACKGROUND: A single nonfunctional chimeric gene with its 5' and 3' ends corresponding to CYP21P and CYP21, respectively, is caused by unequal gene crossover in the CYP21 genes during meiosis. The presence of the chimeric CYP21P/CYP21 molecule can not be detected by conventional PCR methods and therefore may be lost in PCR amplification. This leads to a false result and diagnostic discordance. METHODS: We developed a rapid and direct method to detect a chimeric CYP21P/CYP21 gene that uses a 3'-specific primer for the CYP21 gene and two different 5' primers for both CYP21 and CYP21P to amplify the wild-type CYP21 and the chimeric CYP21P/CYP21 genes. A secondary PCR that can differentiate the chimeric from the wild-type gene was also performed. The PCR product was directly analyzed on agarose gel. RESULTS: After careful titration, we found that earlier failure to detect the chimeric CYP21P/CYP21 gene could be caused by unequal concentrations of two independent alleles as the PCR template or by the lack of primers to amplify chimeric molecules. We successfully amplified the chimeric gene using our improved method. CONCLUSIONS: The chimeric CYP21P/CYP21 is present in a large portion of congenital adrenal hyperplasia patients. By adding a CYP21P/CYP21-specific primer, we were able to amplify and detect both homozygous and heterozygous chimeric genes. Therefore, our new PCR-based assay is a more effective way to analyze congenital adrenal hyperplasia mutations. PMID- 10794741 TI - Analysis of concentration and (13)C enrichment of D-galactose in human plasma. AB - BACKGROUND: A stable-isotope dilution method for the sensitive determination of D galactose in human plasma was established. METHODS: D-[(13)C]Galactose was added to plasma, and the concentration was measured after D-glucose was removed from the plasma by treatment with D-glucose oxidase and the sample was purified by ion exchange chromatography. For gas chromatographic-mass spectrometric analysis, aldononitrile pentaacetate derivatives were prepared. Monitoring of the [MH 60](+) ion intensities at m/z 328, 329, and 334 in the positive chemical ionization mode allowed the assessment of 1-(12)C-, 1-(13)C-, and U-(13)C(6) labeled D-galactose, respectively. The D-galactose concentration was quantified on the basis of the (13)C-labeled internal standard. RESULTS: The method was linear (range examined, 0.1-5 micromol/L) and of good repeatability in the low and high concentration ranges (within- and between-run CVs <15%). The limit of quantification for plasma D-galactose was <0.02 micromol/L. Measurements in plasma of postabsorptive subjects yielded D-galactose concentrations (mean +/- SD) of 0.12 +/- 0.03 (n = 16), 0.11 +/- 0.04 (n = 15), 1.44 +/- 0.54 (n = 10), and 0.17 +/- 0.07 (n = 5) micromol/L in healthy adults, diabetic patients, patients with classical galactosemia, and obligate heterozygous parents thereof, respectively. These data were considerably lower (3- to 18-fold) than the values of a conventional enzymatic assay. The procedure was also applied successfully in a stable-isotope turnover study to evaluate endogenous D-galactose formation. CONCLUSIONS: The present findings establish that detection of D-galactose from endogenous sources is feasible in human plasma and show that erroneously high results may be obtained by enzymatic methods. PMID- 10794742 TI - Detection of rare mutant alleles by restriction endonuclease-mediated selective PCR: assay design and optimization. AB - BACKGROUND: Restriction endonuclease-mediated selective (REMS)-PCR, allows detection of point mutations, deletions, and insertions. Reactions require concurrent activity of a restriction endonuclease (RE) and a DNA polymerase, both of which must be sufficiently thermostable to retain activity during thermocycling. The inclusion of the RE in REMS-PCR inhibits amplification of sequences containing the RE recognition site, thus producing selective amplification of sequences that lack the RE site. METHODS: Assays were used that allowed the selection of conditions that produce concurrent RE/DNA polymerase activity. The RE thermostability assay involved thermocycling a RE under various conditions and assessing residual cleavage activity at various time points. Conditions found to preserve RE activity during thermocyling were then tested for their compatibility with DNA polymerase-mediated PCR. RESULTS: A range of conditions that preserve activity of the RE BstNI over 30 cycles of PCR was identified. A subset of these conditions was subsequently found to mediate specific amplification using Taq DNA polymerase. These conditions were used to develop a REMS-PCR protocol for the detection of mutations at codon 12 of the K ras gene. This protocol allowed the detection of 1 mutant allele in a background of 1000 wild-type alleles. The presence of primer sets for RE and PCR control amplicons provided unambiguous assessment of mutant status. CONCLUSION: Implementation of the assays described may facilitate development of REMS-PCR assays targeted to other loci associated with disease. PMID- 10794743 TI - DzyNA-PCR: use of DNAzymes to detect and quantify nucleic acid sequences in a real-time fluorescent format. AB - BACKGROUND: DzyNA-PCR is a general strategy for the detection and quantification of specific genetic sequences associated with disease or the presence of foreign agents. The method allows homogeneous gene amplification coupled with signal detection in a single closed vessel. METHODS: The strategy involves in vitro amplification of genetic sequences using a DzyNA primer that harbors the complementary (antisense) sequence of a 10-23 DNAzyme. During amplification, amplicons are produced that contain active (sense) copies of DNAzymes that cleave a reporter substrate included in the reaction mixture. The accumulation of amplicons during PCR can be monitored in real time by changes in fluorescence produced by separation of fluoro/quencher dye molecules incorporated into opposite sides of a DNAzyme cleavage site within the reporter substrate. The DNAzyme and reporter substrate sequences can be generic and hence can be adapted for use with primer sets targeting various genes or transcripts. RESULTS: Experiments using K-ras plasmid as template demonstrated that DzyNA-PCR allows quantification of DNA over at least six orders of magnitude (r = 0.992). Studies with human genomic DNA demonstrated the ability to resolve as little as twofold differences in the amount of starting template. DzyNA-PCR allowed the detection of 10 or fewer copies of the target. The clinical utility of the assay was demonstrated using DzyNA-PCR to analyze DNA that was isolated from human serum. CONCLUSION: DzyNA-PCR is a simple, rapid, and sensitive technique for homogeneous amplification and quantification of nucleic acids in clinical specimens. PMID- 10794744 TI - Rapid detection of point mutations by fluorescence resonance energy transfer and probe melting curves in Candida species. AB - BACKGROUND: The LightCycler(TM) combines rapid amplification of nucleic acids in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection of point mutations in various settings, such as drug resistance and hereditary diseases. Point mutations leading to an altered structure of lanosteroldemethylase, the target enzyme of the fungistatic azoles, are an important mechanism of acquired resistance in Candida albicans. METHODS: We screened 13 fluconazole-resistant C. albicans and 21 fluconazole-resistant C. tropicalis strains (minimum inhibitory concentration >128 mg/L), isolated from patients with AIDS, for the presence of defined point mutations by comparing conventional cycle sequencing with a newly designed LightCycler-based assay. RESULTS: In C. tropicalis, 5 of 21 isolates showed the wild-type sequence, and 8 of 21 showed the homozygous nucleotide exchange thymine to cytosine at position 1554 (T1554C). A heterozygous genotype was detected in 8 of 21 isolates by the LightCycler, but in only 3 of 21 isolates by conventional cycle sequencing. In 2 of 13 C. albicans isolates, a homozygous point mutation leading to an amino acid exchange at position 464 (glycine to serine) was detected in both assays. CONCLUSION: The LightCycler technique offers standardized, fast, sensitive, and reproducible detection of point mutations in different Candida spp. PMID- 10794745 TI - ELISA for urinary trehalase with monoclonal antibodies: a technique for assessment of renal tubular damage. AB - BACKGROUND: alpha,alpha-Trehalase, located on renal proximal tubules, is a glycoprotein that hydrolyses alpha,alpha-trehalose to two glucose molecules. Urinary trehalase reflects damage to renal proximal tubules, but its activity has not been measured routinely because measurement of catalytic activity is rather complicated and because conventional assays for enzyme activity might not reflect all of the trehalase protein because of enzyme inactivation in urinary samples. METHODS: We established novel monoclonal antibodies for human trehalase and a sandwich ELISA for quantification of urinary trehalase. We determined the urinary trehalase protein concentration with this ELISA and trehalase catalytic activity, and the results of these two methods were compared. RESULTS: The ELISA system was more sensitive than the detection of enzyme activity and could detect a subtle difference in the amount of trehalase present in renal diseases. The within- and between-assay CVs in the ELISA were 6.7-7.6% and 6.2-8.2%, respectively. Highly significant increases in both the quantity and activity were seen in patients with nephrotic syndrome (acute phase), Lowe syndrome, and Dent disease. The quantities were 70- to 200-fold greater, whereas enzyme activities were, at most, 10-fold higher than those of control subjects. In the detection of small amounts of trehalase in patients with chronic glomerulonephritis and renal anomalies, quantities were better than enzyme activities. CONCLUSIONS: We have established an ELISA system for quantification of urinary trehalase that uses novel monoclonal antibodies. Our ELISA system is simpler and more sensitive than a conventional activity assay and reflects trehalase protein. This ELISA can be a useful as a common tool for clinical assessment of renal proximal tubular damage. PMID- 10794746 TI - Routine alpha-amylase assay using protected 4-nitrophenyl-1, 4-alpha-D maltoheptaoside and a novel alpha-glucosidase. AB - BACKGROUND: In contrast to numerous methods for measuring alpha-amylase activity, the approved IFCC reference method offers an invariable time-independent constant product pattern, thus avoiding possibly changing stoichiometric calculations. However, reference methods do not lend themselves to routine use, so that such methods need to be modified. METHODS: Ethylidene-blocked 4 nitrophenylmaltoheptaoside (EPS-G7) is degraded to glucose and 4-nitrophenol in a coupled assay with a bacterial alpha-glucosidase under the following measurement conditions: 3.5 mmol/L EPS-G7, 7.1 kU/L alpha-glucosidase, 70 mmol/L sodium chloride, 1 mmol/L calcium chloride, 50 mmol/L HEPES, pH 7.15, at 37 degrees C. The increase of absorbance is continuously monitored for 3 min at 405 nm after a lag phase of 2 min. RESULTS: Catalytic concentrations up to 15-fold higher than the upper reference limit can be determined without dilution. Precision studies in manual performance show CVs of 1.4-2. 6% (within-run) and 1.9-2.8% (day-to day). There was no interference from 100 mmol/L glucose, 30 mmol/L triacylglycerols, 610 micromol/L bilirubin, and 2.95 g/L hemoglobin. The method closely correlates with other chromogenic assays. The preliminary 0.95 reference interval for adults, not dependent on age and sex, is 33.6-96.2 U/L. CONCLUSION: The procedure is a robust adaptation of the reference method to routine use at 37 degrees C with increased sensitivity, fewer interferences, and reduced cost. PMID- 10794747 TI - Cardiac troponins I and T are biological markers of left ventricular dysfunction in septic shock. AB - BACKGROUND: Cardiac depression in severe sepsis and septic shock is characterized by left ventricular (LV) failure. To date, it is unclear whether clinically unrecognized myocardial cell injury accompanies, causes, or results from this decreased cardiac performance. We therefore studied the relationship between cardiac troponin I (cTnI) and T (cTnT) and LV dysfunction in early septic shock. METHODS: Forty-six patients were consecutively enrolled, fluid-resuscitated, and treated with catecholamines. Cardiac markers were measured at study entry and after 24 and 48 h. LV function was assessed by two-dimensional transesophageal echocardiography. RESULTS: Increased plasma concentrations of cTnI (>/=0.4 microgram/L) and cTnT (>/=0.1 microgram/L) were found in 50% and 36%, respectively, of the patients at one or more time points. cTnI and cTnT were significantly correlated (r = 0.847; P <0.0001). Compared with cTnI-negative patients, cTnI-positive subjects were older, presented higher Acute Physiology and Chronic Health Evaluation II scores at diagnosis, and tended to have a worse survival rate and a more frequent history of arterial hypertension or previous myocardial infarction. In contrast, the two groups did not differ in type of infection or pathogen, or in dose and type of catecholamine administered. Continuous electrocardiographic monitoring in all patients and autopsy in 12 nonsurvivors did not disclose the occurrence of acute ischemia during the first 48 h of observation. LV dysfunction was strongly associated with cTnI positivity (78% vs 9% in cTnI-negative patients; P <0.001). In multiple regression analysis, both cTnI and cTnT were exclusively associated with LV dysfunction (P <0.0001). CONCLUSIONS: These findings suggest that in septic shock, clinically unrecognized myocardial cell injury is a marker of LV dysfunction. The latter condition tends to occur more often in severely ill older patients with underlying cardiovascular disease. Further studies are needed to determine the extent to which myocardial damage is a cause or a consequence of LV dysfunction. PMID- 10794748 TI - Dual-label time-resolved immunofluorometric assay of free and total prostate specific antigen based on recombinant Fab fragments. AB - BACKGROUND: Recombinant Fab fragments are attractive as reagents for novel sandwich immunoassays, but no such assays have been described. We developed a dual-label two-site immunoassay based entirely on recombinant Fab fragments and compared it to the same assay with intact monoclonal antibodies. METHODS: The capture Fab fragment, which binds free prostate-specific antigen (PSA) and PSA in complex with alpha(1)-antichymotrypsin on an equimolar basis, is site specifically biotinylated and attached to the solid phase in streptavidin-coated microtitration wells. The Fab fragment that detects only free PSA is site specifically labeled with a fluorescent europium chelate, and the Fab fragment that detects both free and serpin-complexed PSA in an equimolar fashion is labeled with a fluorescent terbium chelate. Time-resolved fluorescence is used to measure both europium and terbium signals in one well. RESULTS: The detection limits of the assay (mean + 3 SD of zero calibrator) were 0.043 and 0.28 microgram/L, respectively, for free and total PSA. The within-run and day-to-day CVs were 2-11% and 4-10%, respectively. Mean recoveries were 93% and 98% in female and male sera, respectively. Compared with the commercial ProStatus PSA Free/Total Assay, the intercepts of the regression equations (r >0. 99) were not significantly different from zero, and the slopes were 0.95-1.01. In one female serum sample, PSA was falsely increased with the monoclonal assay but was undetectable with the recombinant assay. CONCLUSIONS: The performance of this novel assay based on recombinant components is comparable to a conventional assay based on monoclonal antibodies. The more complete control of the essential characteristics of site-specifically derivatized recombinant Fab fragments will be valuable for the design of miniaturized and multianalyte assay concepts where correct antibody orientation, density, and capacity as well as uncompromised binding affinity are required. PMID- 10794749 TI - Remnant-like particle-cholesterol concentrations in patients with type 2 diabetes mellitus and end-stage renal disease. AB - BACKGROUND: Lipid abnormalities contribute significantly to the increased risk of cardiovascular disease in diabetic and end-stage renal disease (ESRD) patients. Accumulating evidence supports a proatherogenic role for remnant lipoproteins. Thus, the aim of the present study was to compare remnant-like particle cholesterol (RLP-C) in type 2 diabetic and ESRD patients with age- and gender matched controls. METHODS: Using an immunoaffinity assay, we measured RLP-C concentrations in 48 type 2 diabetic patients with (n = 24) and without (n = 24) macrovascular complications, and 24 age- and gender-matched controls, as well as in 38 ESRD patients on hemodialysis (n = 19) and peritoneal dialysis (n = 19), and 19 age- and gender-matched controls. RESULTS: RLP-C correlated significantly with plasma triglycerides (TGs; r = 0.8). When compared with controls, RLP-C concentrations were significantly higher in type 2 diabetic patients with and without macrovascular complications (median, 0.22 and 0.17 mmol/L vs 0.14 mmol/L; P <0.0002 and <0.01, respectively); diabetic patients with macrovascular complications also had significantly higher RLP-C than diabetic patients without macrovascular complications (P <0.05). However, when RLP-C/TG ratios were computed, only diabetic patients with macrovascular complications showed significantly higher RLP-C/TG ratios compared with controls (P <0.05). Regarding ESRD, RLP-C concentrations were significantly increased in patients on both hemodialysis and peritoneal dialysis compared with controls (median, 0.23 and 0.21 mmol/L vs 0.13 mmol/L; P <0.0001). Whereas RLP-C was increased in ESRD patients on hemodialysis with TGs <2.26 mmol/L compared with controls, RLP-C/TG ratios were not significantly increased in these patients. CONCLUSIONS: Type 2 diabetic patients with macrovascular disease demonstrated increased RLP-C and RLP C/TG ratios, whereas ESRD patients showed only increased RLP-C concentrations. PMID- 10794750 TI - Proteasome inhibition measurements: clinical application. AB - BACKGROUND: PS-341, a selective inhibitor of the proteasome, currently is under evaluation as an anticancer agent in multiple phase I clinical trials. In animal model studies, PS-341 was rapidly removed from the vascular compartment and distributed widely, quickly approaching the limits of detection. An accurate pharmacodynamic assay has been developed as an alternative or complement to pharmacokinetic measurements. METHODS: Fluorogenic kinetic assays for both the chymotryptic and tryptic activities of the proteasome have been optimized for both whole blood and blood cells. Using the ratio of these activities and the catalytic mechanism of the proteasome, we developed a novel method of calculating percentage of inhibition, using two structurally unrelated inhibitors (PS-341 and lactacystin). RESULTS: This ratio method was demonstrated to be sensitive (detection limit of 13% inhibition with 10 microgram of cell lysate), specific to the proteasome (PS-341 provides >98% inhibition), accurate (112% analyte recovery), and precise (0% +/- 5% inhibition at 0 nmol/L PS-341 and 74.5% +/- 1.7% inhibition at 200 nmol/L PS-341). Using these assays, we found that both erythrocytes and leukocytes contain proteasome at 3 micromol/L. Pharmacodynamic results for PS-341 obtained from the whole-blood ratio method were comparable to those using leukocytes determined by another method. CONCLUSIONS: The described assay provides a reliable method for studying the pharmacodynamics of proteasome inhibitors and is now in use in concurrent phase I clinical trials with PS-341. PMID- 10794751 TI - Short- and long-term effects of ibandronate treatment on bone turnover in Paget disease of bone. AB - BACKGROUND: In Paget disease of bone (PD), serum total alkaline phosphatase (TAP) is a valid marker of disease activity. The aim of the present longitudinal study was to compare TAP with new and potentially more specific markers of bone turnover in bisphosphonate-treated patients with PD. METHODS: Twenty patients with active PD were studied before and after treatment with 2 mg of intravenous ibandronate over a period of 12 months. TAP (by colorimetry), serum bone-specific alkaline phosphatase (BAP; by enzyme immunoassay), serum osteocalcin (OC; by ELISA), serum bone sialoprotein (BSP; by RIA), and urinary total pyridinoline (PYD; by HPLC) and deoxypyridinoline (DPD; by HPLC) were measured as markers of bone turnover. RESULTS: Before treatment, TAP, BAP, and BSP were increased in all 20 patients, whereas OC was increased in 10, PYD in 13, and DPD in 15 patients. Three months post treatment, nine patients showed normalized TAP values, and a >/=25% re-increase (i.e. , relapse) was observed in all patients after 12 months. A normalization of BAP was achieved in six patients only. No significant changes were found for OC. BSP was decreased significantly at 24 h, and DPD at 48 h post treatment. A normalization of BSP was found in 8, of PYD in 18, and of DPD in 16 cases. Both PYD and DPD increased significantly from 9 months post treatment onward. CONCLUSIONS: Most markers of bone turnover show similar long-term changes after treatment of active PD with ibandronate. With regard to cost-effectiveness and assay performance, TAP remains the marker of choice in therapeutic monitoring of PD. However, more specific markers may improve the biochemical assessment of PD in certain situations. PMID- 10794752 TI - The plasma homocysteine concentration is better than that of serum methylmalonic acid as a marker for sociopsychological performance in a psychogeriatric population. AB - BACKGROUND: Cobalamin/folate deficiency in elderly subjects may lead to psychiatric symptoms, but more often it increases the severity of various organic and nonorganic mental diseases. A major clinical problem, however, is the uncertainty and controversy concerning biochemical markers of cobalamin/folate deficiency to be used in the diagnostic evaluation of suspected cobalamin/folate deficiency. METHODS: We measured plasma homocysteine (tHcy), blood folate, serum methylmalonic acid, and serum cobalamin in 80 psychogeriatric patients (age, 77.3 +/- 8.6 years) and 50 controls (age, 76.1 +/- 8. 0 years). We assessed associations of these tests with measures of cognitive and behavior performance by use of regression analyses. RESULTS: Plasma tHcy was increased in 45% of the psychogeriatric population (mean, 20.5 +/- 9.2 micromol/L vs 15.3 +/- 4.7 micromol/L for controls; P <0.01). Plasma tHcy correlated with severity of dementia (r = 0.36; P <0.01), the Katz ADL index (r = 0.29; P <0.05), the Berger scale (r = 0.29; P <0.05), and the score of symptoms (r = 0.39; P <0.001) in the psychogeriatric population. Similarly, blood folate was significantly correlated with these measures, but the concentrations of serum cobalamin and methylmalonic acid were not. In a stepwise multiple regression analysis including the biochemical markers, tHcy was the only significant predictor of the severity of dementia (r(2) = 0.11; P <0.01) and the score of symptoms (r(2) = 0.16; P <0.001). CONCLUSION: Plasma tHcy is the best marker of those measured to investigate suspected tissue deficiency of cobalamin/folate. PMID- 10794753 TI - Influence of glomerular filtration rate on non-(1-84) parathyroid hormone (PTH) detected by intact PTH assays. AB - BACKGROUND: Commercial intact parathyroid hormone (I-PTH) assays detect molecular form(s) of human PTH, non-(1-84) PTH, different from the 84-amino acid native molecule. These molecular form(s) accumulate in hemodialyzed patients. We investigated the importance of non-(1-84) PTH in the interpretation of the increased I-PTH in progressive renal failure. METHODS: Five groups were studied: 26 healthy individuals, 12 hemodialyzed patients, and 31 patients with progressive renal failure subdivided according to their glomerular filtration rate (GFR) into 11 with a GFR between 60 and 100 mL. min(-1). 1.73 m(-2), 12 with a GFR between 30 and 60 mL. min(-1). 1.73 m(-2), and 8 with a GFR between 5 and 30 mL. min(-1). 1.73 m(-2). We evaluated indicators of calcium and phosphorus metabolism and creatinine clearance (CrCl) in the progressive renal failure groups, and the HPLC profile of I-PTH and C-terminal PTH in all groups. RESULTS: Only patients with a GFR <30 mL. min(-1). 1.73 m(-2) and hemodialyzed patients had decreased Ca(2+) and 1,25-dihydroxyvitamin D, and increased phosphate. In patients with progressive renal failure, I-PTH was related to Ca(2+) (r = -0.66; P <0.0001), CrCl (r = -0.61; P <0.001), 1,25-dihydroxyvitamin D (r = -0.40; P <0.05), and 25-hydroxyvitamin D (r = -0.49; P <0.01) by simple linear regression. The importance of non-(1-84) PTH in the composition of I-PTH increased with each GFR decrease, being 21% in healthy individuals, 32% in progressive renal failure patients with a GFR <30 mL. min(-1). 1.73 m(-2), and 50% in hemodialyzed patients, with PTH(1-84) making up the difference. CONCLUSIONS: As I-PTH increases progressively with GFR decrease, part of the increase is associated with the accumulation of non-(1-84) PTH, particularly when the GFR is <30 mL. min(-1). 1.73 m(-2). Concentrations of I-PTH 1.6-fold higher than in healthy individuals are necessary in hemodialyzed patients to achieve PTH(1-84) concentrations similar to those in the absence of renal failure. PMID- 10794754 TI - Vitamin B(1) status assessed by direct measurement of thiamin pyrophosphate in erythrocytes or whole blood by HPLC: comparison with erythrocyte transketolase activation assay. AB - BACKGROUND: The concentration of thiamin diphosphate (TDP) in erythrocytes is a useful index of thiamin status. We describe an HPLC method for TDP and its results in patients at risk of thiamin deficiency. METHODS: We used reversed phase HPLC with postcolumn derivatization with alkaline potassium ferricyanide and fluorescence detection. Samples were deproteinized and injected directly onto a C(18) column. TDP concentrations in erythrocytes were compared with those in whole blood. Reference intervals for erythrocyte TDP (n = 147; 79 males and 68 females; mean age, 54 years) and whole blood TDP (n = 124; 68 males and 56 females; mean age, 54 years) were determined in an apparently healthy population. We compared erythrocyte TDP with results of the erythrocyte transketolase activation test in 63 patients who were considered at risk of thiamin deficiency. RESULTS: The method was linear to at least 200 microgram/L. The between-run CV was <8%. The lower limit of quantification for both whole blood and packed erythrocytes was 300 pg on column with a detection limit of 130 pg on column. Recovery of TDP from blood samples was >90%. TDP in erythrocytes correlated strongly with that in whole blood (r = 0.97). Reference intervals for erythrocyte and whole blood TDP were 280-590 ng/g hemoglobin and 275-675 ng/g hemoglobin, respectively. Of the 63 patients suspected of thiamin deficiency, 46 were normal by both TDP and activation tests, 13 were deficient by both tests, 1 was deficient by the activation test but had normal erythrocyte TDP concentrations, and 4 were normal by the activation test but had low TDP. CONCLUSIONS: The HPLC method is precise and yields results similar to the erythrocyte activation assay. PMID- 10794755 TI - Improved molecular diagnosis of hereditary hemochromatosis using a DNA enzyme immunoassay. PMID- 10794756 TI - Correlation of serum concentrations of cystatin C and creatinine to inulin clearance in liver cirrhosis. PMID- 10794757 TI - Semiautomated PCR-single-strand conformation polymorphism method for detection of a novel sequence polymorphism (Ile1000Val) in human alpha(2)-macroglobulin. PMID- 10794758 TI - Role of heart-type fatty acid-binding protein in early detection of acute myocardial infarction. PMID- 10794759 TI - Serum transferrin receptor and erythrocyte zinc protoporphyrin in patients with anemia. PMID- 10794760 TI - Evaluation of the apo E genotyping kit on the LightCycler. PMID- 10794761 TI - Serum transferrin receptor concentrations during normal pregnancy. PMID- 10794762 TI - Assay for free and total carnitine in human plasma using tandem mass spectrometry. PMID- 10794763 TI - Detection of apoptotic fetal cells in plasma of pregnant women. PMID- 10794764 TI - Distribution of selenium in plasma of French women: relation to age and selenium status. PMID- 10794765 TI - Stability of clinical chemistry analytes in blood collection devices. PMID- 10794766 TI - Efavirenz interference in estradiol ELISA assay. PMID- 10794767 TI - Thiobarbiturates interfere with the Dade Behring aca ammonia test. PMID- 10794768 TI - Interference of iodinated contrast media in serum capillary zone electrophoresis. PMID- 10794769 TI - Stability of clinical chemistry analytes in blood collection devices PMID- 10794770 TI - National center for infectious diseases (NCID). URL: www.cdc. gov/ncidod/ PMID- 10794771 TI - Laboratory automation: smart strategies and practical applications. PMID- 10794772 TI - Development of standards for laboratory automation. AB - In clinical laboratories, the installation of total laboratory automation systems and/or modular systems has grown dramatically in the 1990s, particularly in the US, Japan, and Europe. As the number of installations and level of interest grew, several individuals and corporations active in the automation field recognized that the development of prospective standards might enable customers of such systems or equipment to purchase analyzers, automation systems or devices, and software from different vendors and retain interconnectivity of such equipment. These individuals also believed that the total market for automation systems and equipment would be significantly greater with standards than without standards, especially if customers were not forced to purchase everything from one vendor, and that there might be competitive pricing and new technology fostered via the standards. This early interest in standards development led to the initiation of a program by NCCLS in 1996 to develop prospective standards for laboratory automation. Part of the NCCLS effort has involved interaction and cooperation with other standards organizations in the US and other countries. This report describes the current status of the development of prospective standards for laboratory automation by NCCLS and the relationship of those standards to those of other standards organizations. PMID- 10794773 TI - The role of total laboratory automation in a consolidated laboratory network. AB - BACKGROUND: In an effort to reduce overall laboratory costs and improve overall laboratory efficiencies at all of its network hospitals, the North Shore-Long Island Health System recently established a Consolidated Laboratory Network with a Core Laboratory at its center. METHODS: We established and implemented a centralized Core Laboratory designed around the Roche/Hitachi CLAS Total Laboratory Automation system to perform the general and esoteric laboratory testing throughout the system in a timely and cost-effective fashion. All remaining STAT testing will be performed within the Rapid Response Laboratories (RRLs) at each of the system's hospitals. RESULTS: Results for this laboratory consolidation and implementation effort demonstrated a decrease in labor costs and improved turnaround time (TAT) at the core laboratory. Anticipated system savings are approximately $2.7 million. TATs averaged 1.3 h within the Core Laboratory and less than 30 min in the RRLs. CONCLUSIONS: When properly implemented, automation systems can reduce overall laboratory expenses, enhance patient services, and address the overall concerns facing the laboratory today: job satisfaction, decreased length of stay, and safety. The financial savings realized are primarily a result of labor reductions. PMID- 10794774 TI - Developing an automation concept that is right for your laboratory. AB - BACKGROUND: Trends in laboratory automation and critical project principles and design concepts are presented. APPROACH: MDS AutoLab technology development and automation projects were reviewed. Successful methods and approaches were extracted. ISSUES: Continued pressure on the laboratory to reduce costs and increase productivity has catalyzed dramatic development in laboratory automation. Today, laboratories can choose from a wide range of options. The most effective choices are not always the most obvious and will not be the same for all laboratories. Laboratory automation projects are highly complex and must be planned and managed across clinical, technical, operational, financial, and human dimensions. CONCLUSIONS: Success requires excellent communications, an understanding of the risks and barriers, and a dedicated team supported by strong champions throughout the organization. The automation project team will need to use a variety of skills and techniques to evaluate and reengineer processes to identify the highest value targets for automation. PMID- 10794775 TI - Laboratory automation: trajectory, technology, and tactics. AB - Laboratory automation is in its infancy, following a path parallel to the development of laboratory information systems in the late 1970s and early 1980s. Changes on the horizon in healthcare and clinical laboratory service that affect the delivery of laboratory results include the increasing age of the population in North America, the implementation of the Balanced Budget Act (1997), and the creation of disease management companies. Major technology drivers include outcomes optimization and phenotypically targeted drugs. Constant cost pressures in the clinical laboratory have forced diagnostic manufacturers into less than optimal profitability states. Laboratory automation can be a tool for the improvement of laboratory services and may decrease costs. The key to improvement of laboratory services is implementation of the correct automation technology. The design of this technology should be driven by required functionality. Automation design issues should be centered on the understanding of the laboratory and its relationship to healthcare delivery and the business and operational processes in the clinical laboratory. Automation design philosophy has evolved from a hardware-based approach to a software-based approach. Process control software to support repeat testing, reflex testing, and transportation management, and overall computer-integrated manufacturing approaches to laboratory automation implementation are rapidly expanding areas. It is clear that hardware and software are functionally interdependent and that the interface between the laboratory automation system and the laboratory information system is a key component. The cost-effectiveness of automation solutions suggested by vendors, however, has been difficult to evaluate because the number of automation installations are few and the precision with which operational data have been collected to determine payback is suboptimal. The trend in automation has moved from total laboratory automation to a modular approach, from a hardware-driven system to process control, from a one-of-a-kind novelty toward a standardized product, and from an in vitro diagnostics novelty to a marketing tool. Multiple vendors are present in the marketplace, many of whom are in vitro diagnostics manufacturers providing an automation solution coupled with their instruments, whereas others are focused automation companies. Automation technology continues to advance, acceptance continues to climb, and payback and cost justification methods are developing. PMID- 10794776 TI - Modular robotic workcell for coagulation analysis. AB - BACKGROUND: Total laboratory automation (TLA) has been shown to increase laboratory efficiency and quality. However, modular automation is smaller, requires less initial capital, and requires less planning than TLA. We engineered and performed clinical trials on a modular robotic preanalytical workcell for coagulation analysis. METHODS: Timing studies were used to quantify the efficiency of the manual processes and to identify areas in the processing of coagulation specimens where bottlenecks and long waiting periods were encountered. We then designed our modular robotic system to eliminate these bottlenecks. Our robotic modular workcell was engineered to allow a choice of specimen introduction manually, by conveyor, or by mobile robot. Additional timing studies were performed during clinical trials of the robotic system. RESULTS: Prior to automation, the time required for preanalytical processing time was 18-107 min; after automation, it was 45-50 min. Additional improvements in workcell efficiency could be realized when high quality, prelabeled specimens were introduced into the system. CONCLUSION: Compared with manual methods, modular automation provides more predictable variation in specimen processing. PMID- 10794777 TI - Stand-alone automated solutions can enhance laboratory operations. AB - Clinical laboratory automation has developed over the past decade as one means of consolidating testing, reducing costs, and improving the effectiveness of laboratory testing. Most of the developments have been aimed at core clinical laboratory operations, and have primarily addressed preanalytical and analytical processing of traditional specimens arriving in blood collection or similar aliquot tubes. Much less attention has been given to specialized applications such as processing specimens for urine toxicology, and only recently have vendors attacked the problems associated with sorting and maintaining the laboratory's inventory of specimens. This report highlights selected developments in these areas, describes one approach to cost-effective custom platform development, and discusses the advantages and pitfalls to solving problems with laboratory automation. PMID- 10794779 TI - Compiled by david E. Bruns, editor (dbruns@aacc.org) : the clinical chemist PMID- 10794778 TI - Laboratory automation and optimization: the role of architecture. AB - The increasing automation of laboratory equipment has had far-reaching impacts on the organizational structure and spatial requirements of clinical laboratories. This report explores the changing role of the laboratory in the healthcare environment and shows the architectural impact of these changes, both inside and outside of the laboratory space. PMID- 10794781 TI - Embryology of the circulation PMID- 10794780 TI - Management of severe alloimmune thrombocytopenia in the newborn. PMID- 10794782 TI - Antenatal treatment of a mother bearing a fetus with congenital adrenal hyperplasia. PMID- 10794783 TI - Superior vena cava flow in newborn infants: a novel marker of systemic blood flow. AB - BACKGROUND: Ventricular outputs cannot be used to assess systemic blood flow in preterm infants because they are confounded by shunts through the ductus arteriosus and atrial septum. However, flow measurements in the superior vena cava (SVC) can assess blood returning from the upper body and brain. OBJECTIVES: To describe a Doppler echocardiographic technique that measures blood flow in the SVC, to test its reproducibility, and to establish normal ranges. DESIGN: SVC flow was assessed together with right ventricular output and atrial or ductal shunting. Normal range was established in 14 infants born after 36 weeks' gestation (2 measurements taken in the first 48 hours) and 25 uncomplicated infants born before 30 weeks (4 measurements taken in the first 48 hours). Intra observer and interobserver variability were tested in 20 preterm infants. RESULTS: In 14 infants born after 36 weeks, median SVC flow rose from 76 ml/kg/min on day 1 to 93 ml/kg/min on day 2; in 25 uncomplicated very preterm infants, it rose from 62 ml/kg/min at 5 hours to 86 ml/kg/min at 48 hours. The lowest SVC flow for the preterm babies rose from 30 ml/kg/min at 5 hours to 46 ml/kg/min by 48 hours. Median intra-observer and interobserver variability were 8. 1% and 14%, respectively. In preterm babies with a closed duct, SVC flow was a mean of 37% of left ventricular output and the two measures correlated significantly. CONCLUSIONS: This technique can assess blood flow from the upper body, including the brain, in the crucial early postnatal period, and might allow more accurate assessment of the status of systemic blood flow and response to treatment. PMID- 10794784 TI - Low superior vena cava flow and intraventricular haemorrhage in preterm infants. AB - OBJECTIVES: To document the incidence, timing, degree, and associations of systemic hypoperfusion in the preterm infant and to explore the temporal relation between low systemic blood flow and the development of intraventricular haemorrhage (IVH). STUDY DESIGN: 126 babies born before 30 weeks' gestation (mean 27 weeks, mean body weight 991 g) were studied with Doppler echocardiography and cerebral ultrasound at 5, 12, 24, and 48 hours of age. Superior vena cava (SVC) flow was assessed by Doppler echocardiography as the primary measure of systemic blood flow returning from the upper body and brain. Other measures included colour Doppler diameters of ductal and atrial shunts, as well as Doppler assessment of shunt direction and velocity, and right and left ventricular outputs. Upper body vascular resistance was calculated from mean blood pressure and SVC flow. RESULTS: SVC flow below the range recorded in well preterm babies was common in the first 24 hours (48 (38%) babies), becoming significantly less common by 48 hours (6 (5%) babies). These low flows were significantly associated with lower gestation, higher upper body vascular resistance, larger diameter ductal shunts, and higher mean airway pressure. Babies whose mothers had received antihypertensives had significantly higher SVC flow during the first 24 hours. Early IVH was already present in 9 babies at 5 hours of age. Normal SVC flows were seen in these babies except in 3 with IVH, which later extended, who all had SVC flow below the normal range at 5 and/or 12 hours. Eight of these 9 babies were delivered vaginally. Late IVH developed in 18 babies. 13 of 14 babies with grade 2 to 4 IVH had SVC flow below the normal range before development of an IVH. Two of 4 babies with grade 1 IVH also had SVC flow below the normal range before developing IVH, and the other 2 had SVC flow in the low normal range. In all, IVH was first seen after the SVC flow had improved, and the grade of IVH related significantly to the severity and duration of low SVC flow. The 9 babies who had SVC flow below the normal range and did not develop IVH or periventricular leucomalacia were considerably more mature (median gestation 28 v 25 weeks). CONCLUSIONS: Low SVC flow may result from an immature myocardium struggling to adapt to increased extrauterine vascular resistances. Critically low flow occurs when this is compounded by high mean airway pressure and large ductal shunts out of the systemic circulation. Late IVH is strongly associated with these low flow states and occurs as perfusion improves. PMID- 10794785 TI - A preliminary study of the application of the transductal velocity ratio for assessing persistent ductus arteriosus. AB - OBJECTIVE: To compare the transductal velocity ratio (TVR) of the persistent ductus arteriosus (PDA) with other echocardiographic criteria for haemodynamic significance of a PDA. METHODS: This was a prospective study (from January 1997 to August 1998) in the nurseries of the Royal Women's Hospital, Melbourne. Infants with a clinically suspected PDA were eligible and included if the echocardiogram showed a PDA with a structurally normal heart and the TVR had been measured. The PDA was assessed for evidence of left heart dilatation, the presence of reverse or absent diastolic flow in the descending aorta, the pattern of Doppler flow velocity waveform in the ductus arteriosus, and subjective assessment of ductal diameter on the real time image. The peak systolic velocity (PSV) was obtained from the pulmonary and aortic ends of the PDA, and the TVR calculated by dividing the PSV at the pulmonary end by the PSV at the aortic end. RESULTS: Forty two infants had 59 echocardiographs with their TVR calculated. Mean (SD) birth weight was 1008 (362) g. Mean (SD) gestational age at birth was 27.4 (2.2) weeks with a mean (SD) corrected gestational age of 28.7 (2.7) weeks. The mean TVR was decreased in those infants with a high left atrial diameter/aortic diameter (LA/Ao) ratio (1.9 v 2.8, p = 0.0032) or reverse/absent diastolic flow in the descending aorta (2.1 v 3.0, p = 0.02). This difference was greater if those two criteria were combined (1.7 v 3.4, p = 0.0027). The mean TVR was decreased in infants with a wide open duct seen on two dimensional imaging (1.5 v 3.0, p < 0.0001) or pulsatile flow seen on pulsed Doppler in the PDA (1.9 v 3.4, p = 0.0001). The LA/Ao and left ventricle internal diameter/aortic diameter (LVIDd/Ao) ratios were higher in the group with a TVR < 1.8 than in the other two groups; these differences were statistically significant. CONCLUSIONS: The TVR as a measure of the degree of constriction of a PDA is associated with other echocardiographic criteria for a haemodynamically significant PDA. A low TVR (signifying a poorly constricted duct) is associated with echocardiographic features of a significant left to right shunt, and vice versa. Further research is required to determine the usefulness of the TVR in predicting closure or likely continuing patency of a PDA and the need for treatment. PMID- 10794786 TI - Antenatal and perinatal predictors of infant mortality in rural Malawi. AB - BACKGROUND: The slow pace in the reduction of infant mortality in sub-Saharan Africa has partially been attributed to the epidemic of human immunodeficiency virus (HIV) infection. To facilitate early interventions, antenatal and perinatal predictors of 1st year mortality were identified in a rural community in southern Malawi. METHODS: A cohort of 733 live born infants was studied prospectively from approximately 24 gestation weeks onwards. Univariate analysis was used to determine relative risks for infant mortality after selected antenatal and perinatal exposures. Multivariate modelling was used to control for potential confounders. FINDINGS: The infant mortality rate was 136 deaths/1000 live births. Among singleton newborns, the strongest antenatal and perinatal predictors of mortality were birth between May and July, maternal primiparity, birth before 38th gestation week, and maternal HIV infection. Theoretically, exposure to these variables accounted for 22%, 22%, 17%, and 15% of the population attributable risk for infant mortality, respectively. INTERPRETATION: The HIV epidemic was an important but not the main determinant of infant mortality. Interventions targetting the offspring of primiparous women or infants born between May and July or prevention of prematurity would all have considerable impact on infant survival. PMID- 10794787 TI - Neonatal group B streptococcal infection in South Bedfordshire, 1993-1998. AB - BACKGROUND: Group B streptococcus (GBS) is now the leading cause of neonatal bacterial sepsis in the western world. The incidence of GBS infection in the United States has been determined, and guidelines produced and implemented for the prevention of neonatal infection. Neither incidence nor guidelines are currently established in the United Kingdom. AIM: To define the pattern of neonatal infection within one hospital (Luton and Dunstable Hospital). METHOD: A six year retrospective analysis was performed. RESULT: An incidence of early onset GBS of 1.15 per 1000 deliveries, comparable with that documented in the United States, was found. PMID- 10794789 TI - Thyroid function in very low birthweight infants after intravenous administration of the iodinated contrast medium iopromide. AB - BACKGROUND: Thyroid function disorders have often been observed in preterm infants after intravenous administration of iodinated contrast medium. The effect on thyroid function depends on the dosage, but the choice of the contrast medium may be equally important, as there are appreciable pharmacological differences between them. METHOD: Thyroid function was analysed in 20 very low birthweight infants of gestational age less than 30 weeks after injection of iopromide, a monomeric non-ionic iodinated contrast medium. Levels of free thyroxine and thyroid stimulating hormone were compared with those in 26 control infants. RESULTS: Free thyroxine levels in all study infants ranged from 9.0 to 25.7 pmol/l (days 14-21) and 9.0 to 23.2 pmol/l (days 35-49), and thyroid stimulating hormone levels ranged from 0.13 to 0.26 mU/l (days 14-21) and 0.26 to 11.11 mU/l (days 35-49). These levels were not altered after injection of iopromide. CONCLUSION: The risk of transient hypothyroidism or hyperthyrotropinaemia may be reduced with the use of iopromide compared with other contrast media. PMID- 10794788 TI - Effects of gestation and birth weight on the growth and development of very low birthweight small for gestational age infants: a matched group comparison. AB - AIMS: To investigate the effects of small for gestational age (SGA) in very low birthweight (VLBW) infants on growth and development until the fifth year of life. METHODS: VLBW (< 1500 g) infants, selected from a prospective study, were classified as SGA (n = 115) on the basis of birth weight below the 10th percentile for gestational age and were compared with two groups of appropriate for gestational age (AGA) infants matched according to birth weight (AGA-BW; n = 115) or gestation at birth (AGA-GA; n = 115). Prenatal, perinatal, and postnatal risk factors were recorded, and duration and intensity of treatment were computed from daily assessments. Body weight, length, and head circumference were measured at birth, five and 20 months (corrected for prematurity), and at 56 months. General development was assessed at five and 20 months with the Griffiths scale of babies abilities, and cognitive development at 56 months with the Columbia mental maturity scales, a vocabulary (AWST) and language comprehension test (LSVTA). RESULTS: Significant group differences were found in complications (pregnancy, birth, and neonatal), parity, and multiple birth rate. The AGA-GA group showed most satisfactory growth up to 56 months, with both the AGA-BW and SGA groups lagging behind. The AGA-GA group also scored significantly more highly on all developmental and cognitive tests than the other groups. Developmental test results were similar for the SGA and AGA-BW groups at five and 20 months, but AGA-BW infants (lowest gestation) had lower scores on performance intelligence quotient and language comprehension at 56 months than the SGA group. When prenatal and neonatal complications, parity, and multiple birth were accounted for, group differences in growth remained, but differences in cognitive outcome disappeared after five months. CONCLUSIONS: Being underweight and with a short gestation (SGA and VLBW) leads to poor weight gain and head growth in infancy but does not result in poorer growth than in infants of the same birth weight but shorter gestation (AGA-BW) in the long term. SGA is related to early developmental delay and later language problems; however, neonatal complications may have a larger detrimental effect on long term cognitive development of VLBW infants than whether they are born SGA or AGA. PMID- 10794790 TI - Reference ranges for the linear dimensions of the intracranial ventricles in preterm neonates. AB - AIMS: To establish normal ranges, in preterm infants < 33 weeks' gestation, for measurements of the lateral, third, and fourth ventricles and to assess intra observer and inter-observer reliability. To assess the effect of head position during scanning on lateral ventricle size. To determine whether sex influences ventricle size. METHODS: A prospective study involving infants < 33 weeks' gestational age (GA) at birth. Cranial ultrasound scans were done during the first 3 days of life. Linear dimensions of the anterior horn width and thalamo occipital distance of the lateral ventricles, the width of the third ventricle, and the width and length of the fourth ventricle were measured. Measurements were plotted against GA and reference ranges produced. All measurements were tested for intra-observer and inter-observer reliability. Head position and sex differences were studied. RESULTS: 120 infants with known GA (23(+1) to 32(+6) weeks) had their intracranial ventricles measured. Reference ranges obtained were anterior horn width: 0-2.9 mm; thalamo-occipital distance: 8.7-24.7 mm; third ventricle width: 0-2.6 mm; fourth ventricle width: 3.3-7.4 mm; fourth ventricle length: 2.6-6.9 mm. Dependent and non-dependent lateral ventricles did not differ significantly in size. There was no clinically significant difference in ventricular size between sexes. CONCLUSIONS: Reference ranges for the measurement of the intracranial ventricles in preterm infants from 23 to 33 weeks' GA are provided and can be used in the diagnosis and assessment of ventricular enlargement in preterm infants. All measurements have good intra-observer and inter-observer reliability. Head position at the time of scanning does not influence the asymmetry of the lateral ventricular measurements. The infant's sex does not influence ventricular size. PMID- 10794791 TI - Anoxic ATP depletion in neonatal mice brainstem is prevented by creatine supplementation. AB - BACKGROUND: Sufficient ATP concentrations maintain physiological processes and protect tissue from hypoxic damage. With decreasing oxygen concentration, ATP synthesis relies increasingly on the presence of phosphocreatine. AIM: The effect of exogenously applied creatine on phosphocreatine and ATP concentrations was studied under control and anoxic conditions. METHODS: Pregnant mice were fed orally with creatine monohydrate (2 g/kg body weight/day). Brainstem slices from these mice pups were compared with those from pups of non-creatine supplemented pregnant mice. Measurements were performed under normoxic and anoxic conditions. In addition, brainstem slices from non-creatine treated mice pups were incubated for 3 hours in control artificial cerebrospinal fluid (CSF) (n = 10) or in artificial CSF containing 200 microM creatine (n = 10). ATP and phosphocreatine contents were determined enzymatically in single brainstem slices. RESULTS: ATP concentrations were in the same range in all preparations. However, there was a significant increase of phosphocreatine in the brainstems from pups of creatine fed mice when compared with the brainstems of pups from non-creatine treated mice or in non-incubated brainstems of control animals. After 30 minutes anoxia, ATP as well as phosphocreatine concentrations remained significantly higher in creatine pretreated slices compared with controls. CONCLUSION: The data indicate that exogenous application of creatine is effective in neuroprotection. PMID- 10794792 TI - Therapeutic insulin and hepatic glucose-6-phosphatase activity in preterm infants. AB - BACKGROUND: Hepatic glucose-6-phosphatase activity is low at birth, and in term infants rises rapidly to adult levels. In contrast, in most preterm infants, it remains low postnatally making them vulnerable to repeated hypoglycaemic episodes, resultant cerebral damage, or risk of sudden and unexpected death. AIMS: To investigate the clinical features of preterm infants with low glucose-6 phosphatase enzyme activity to determine the influencing factors. METHODS: Clinical data from 36 preterm infants were correlated by stepwise multiple regression analysis with V(max) of hepatic glucose-6-phosphatase as the dependent variable. RESULTS: The most significant correlation was with the administration of insulin (units/kg/h postnatal life) with lesser effects of respiratory distress syndrome and dopamine administration. The V(max) changes reflected changes in the level of expression of the glucose-6-phosphatase protein. CONCLUSION: In a variety of animal models, hepatic glucose-6-phosphatase levels have been shown to decrease in response to insulin, which also decreases transcription of the glucose-6-phosphatase gene. The association of insulin administration with high levels of hepatic glucose-6-phosphatase activity and protein expression was therefore most unexpected. Results from model systems, or adults, must be extrapolated to the metabolism of preterm infants with caution. PMID- 10794793 TI - Contact lens electroretinography in preterm infants from 32 weeks after conception: a development in current methodology. AB - AIM: To assess the feasibility of using a contact lens electrode to record the electroretinogram (ERG) in preterm infants less than 35 weeks after conception. METHODS: The ERG was recorded from seven very low birthweight preterm infants on a total of 14 occasions using an infant monkey contact lens electrode. Age at recording the first ERG ranged from 23 to 51 days (gestational age 32-34 weeks), and weight ranged upwards from 1100 g. RESULTS: No complications were observed. With advancing age and maturity the dark adapted rod threshold decreased, indicating increased retinal sensitivity. CONCLUSIONS: Contact lens recording of the ERG from extremely small immature preterm infants is a practicable and well tolerated procedure. This method of recording the ERG will enable further evaluation of retinal development in this vulnerable population. PMID- 10794794 TI - Effect of changes in oscillatory amplitude on PaCO(2) and PaO(2) during high frequency oscillatory ventilation. AB - AIMS: To describe the relation between oscillatory amplitude changes and arterial blood gas (ABG) changes in preterm infants receiving high frequency oscillatory ventilation, using a multiparameter intra-arterial sensor (MPIAS). METHODS: Continuous MPIAS ABG data were collected after amplitude changes and stratified according to FIO(2): high (> 0.4) or low (< 0.3). For each amplitude change, the maximum change (from baseline) in PaCO(2) and PaO(2) over the following 30 minutes was determined. In total, 64 oscillatory amplitude changes were measured in 21 infants (median birth weight 1040 g; gestation 27 weeks). RESULTS: All amplitude increases produced PaCO(2) falls (median -0.98 and -1.13 kPa for high and low FIO(2) groups respectively). All amplitude decreases produced PaCO(2) rises (median +0.94 and +1.24 kPa for high and low FIO(2) groups respectively). About 95% of the change in PaCO(2) was completed in 30 minutes. Amplitude changes did not affect PaO(2) when FIO(2) > 0.4. When FIO(2) < 0.3, amplitude increases produced a PaO(2) rise (median = +1.1 kPa; P < 0.001) and amplitude decreases a fall (median = -1.2 kPa; P < 0.001). CONCLUSIONS: After oscillatory amplitude changes, the speed but not the magnitude of the PaCO(2) change is predictable, and a rapid PaO(2) change accompanies the PaCO(2) change in infants with mild lung disease and a low FIO(2). PMID- 10794795 TI - Changes in pulmonary arterial pressure in preterm infants with chronic lung disease. AB - BACKGROUND: Pulmonary arterial pressure (PAP) is raised in preterm infants with respiratory distress syndrome who subsequently develop chronic lung disease. The natural history of pulmonary hypertension in infants with chronic lung disease is unknown. OBJECTIVES: To investigate changes in PAP, assessed non-invasively using Doppler echocardiography, in infants with chronic lung disease during the 1st year of life. METHODS: Serial examinations were performed in infants with chronic lung disease and healthy preterm infants. The Doppler derived acceleration time to right ventricular ejection time ratio (AT/RVET) was calculated from measurements made from the pulmonary artery velocity waveform. RESULTS: A total of 248 examinations were performed in 54 infants with chronic lung disease and 44 healthy preterm infants. The median AT/RVET was significantly lower in infants with chronic lung disease than in healthy preterm infants (0.31 v 0.37). AT/RVET significantly correlated with age corrected for prematurity in both infants with chronic lung disease (r = 0.67) and healthy infants (r = 0.55). There was no significant difference between the rate of change in AT/RVET between the two groups. In infants with chronic lung disease, multivariate analysis showed that AT/RVET was significantly independently associated with age and inversely with duration of supplemental oxygen treatment. Median AT/RVET was significantly lower in infants with chronic lung disease until 40-52 weeks of age corrected for prematurity. CONCLUSIONS: Although PAP falls with increasing age in both infants with chronic lung disease and healthy preterm infants, it remains persistently raised in infants with chronic lung disease until the end of the 1st year of life. PMID- 10794796 TI - Unilateral pleural effusion complicating central venous catheterisation. AB - Acute respiratory distress developed in two preterm babies because of unilateral hydrothorax secondary to the migration of a central venous catheter into the pulmonary vasculature. Prompt recognition of the problem and rapid treatment are essential and life saving. This complication of intravenous alimentation catheters has not been previously reported in the neonatal age group. PMID- 10794797 TI - The fetal and neonatal hypothalamic-pituitary-adrenal axis. PMID- 10794798 TI - Laurent Joubert of Montpellier (1529-82) and his Erreurs Populaires. PMID- 10794799 TI - Genetic immunotherapy for cancer. AB - Genetic immunization refers to treatment strategies where gene transfer methods are used to generate immune responses against cancer. Our growing knowledge of the mechanisms regulating the initiation and maintenance of cytotoxic immune responses has provided the rationale for the design of several genetic immunization strategies. Tumor cells have been gene-modified to express immune stimulatory genes and are then administered as tumor vaccines, in an attempt to overcome tumor cell ignorance by the immune system. With the description of well characterized tumor antigens, multiple strategies have been proposed mainly aimed at optimal tumor antigen presentation by antigen-presenting cells (APC). Among APC, the dendritic cells have been recognized as the most powerful cells in this class, and have become the target for introducing tumor antigen genes to initiate antitumor immune responses. The detailed knowledge of how the immune system can be activated to specifically recognize tumor antigens, and the mechanisms involved in the control of this immune response, provide the basis for modern genetic immunization strategies for cancer treatment. PMID- 10794800 TI - Oral chemotherapeutic agents for colorectal cancer. AB - A number of novel oral chemotherapeutic agents are entering practice or are under development in the United States. Many of these agents display significant clinical activity against colorectal cancer. Many classes of compounds, including fluoropyrimidine analogs, dihydropyrimidine dehydrogenase (DPD) inhibitors, topoisomerase inhibitors, farnesyl transferase inhibitors, and others, are being developed for oral administration. This manuscript describes the progress of clinical development of these agents and also explores the relative merits and challenges of these approaches. Economic issues, patient preference, and patient selection issues surrounding oral chemotherapy for colorectal cancer will also be discussed. PMID- 10794801 TI - MMP-1 is a prognostic marker for hematogenous metastasis of colorectal cancer. AB - BACKGROUND: Degradation of basement membrane and extracellular matrix by matrix metalloproteinases (MMPs) is believed to be an essential step in the complicated process of hematogenous metastasis. MMP-1 is a member of collagenases, a family of MMPs that degrades collagens type I, II, and III, main components of the interstitial stroma. The purpose of this study was to investigate the expression of MMP-1 in colorectal cancer and its correlation with hematogenous metastasis. Patients and Methods. We examined 133 cases of colorectal cancer (Dukes A: 72; Dukes B: 26; Dukes C: 23; Dukes D: 12). Sections were cut from formalin-fixed, paraffin-embedded samples containing the deepest site of cancer invasion and stained immunohistochemically with a monoclonal antibody to MMP-1. According to the area of the tumor that was stained, patients were divided into high- and low MMP-1 expression groups. RESULTS: MMP-1 expression was observed in the cytoplasm of cancer cells, some stromal cells, and a few normal epithelial cells of colonic mucosa. High MMP-1 expression was found in 47 (35.3%) cases and low in 86 (64.7%). Hematogenous metastasis was identified in 14 (29.8%) of high-MMP-1 groups and 12 (13.9%) of low-MMP-1 groups. MMP-1 expression significantly correlated with hematogenous metastasis of colorectal cancer, but no correlation was found between MMP-1 expression and the other clinicopathological features investigated. CONCLUSIONS: MMP-1 expression may be a novel marker for hematogenous metastasis of colorectal cancer, and its inhibition may be a strategy for prevention of metastasis. PMID- 10794802 TI - Non-small cell lung cancer: a study of long-term survival after vinorelbine monotherapy. AB - This study reports the results of 120 patients with inoperable non-small cell lung cancer treated with Navelbine at a dose of 25-30 mg/m(2)/week in a single drug chemotherapy regimen. Surgery was contraindicated due to staging or to concomitant morbidity. Twenty patients achieved survival greater than or equal to 18 months, and one patient obtained exceptional survival of more than 120 months. The mean dose intensity of Navelbine in long-term survivors was 21.61 mg/m(2)/week. Objective response to Navelbine was found by multivariate analysis to be a prognostic factor for survival beyond 18 months. Weight loss of more than 5 kg of corporal weight was an unfavorable prognostic factor in patients with metastatic disease. PMID- 10794803 TI - New drugs and novel targets for treatment of invasive fungal infections in patients with cancer. AB - Invasive fungal infections have emerged as important causes of morbidity and mortality in profoundly immunocompromised patients with cancer. Current treatment strategies for these infections are limited by antifungal resistance, toxicity, drug interactions, and expense. In order to overcome these limitations, new antifungal compounds are being developed, which may improve our therapeutic armamentarium for prevention and treatment of invasive mycoses in high-risk patients with neoplastic diseases. PMID- 10794805 TI - Temozolomide, a novel alkylating agent with activity in the central nervous system, may improve the treatment of advanced metastatic melanoma. AB - Temozolomide (TMZ) is the first new chemotherapy agent to be approved for the treatment of high-grade malignant gliomas in more than 20 years. This novel oral alkylating agent has demonstrated promising activity not only in brain tumors, but in a variety of solid tumors, including malignant melanoma. TMZ is 100% bioavailable when taken orally and, because of its small size and lipophilic properties, it is able to cross the blood-brain barrier. Concentrations in the central nervous system are approximately 30% of plasma concentrations. Once it has entered the central nervous system, TMZ can be spontaneously converted to the active metabolite. These pharmacologic properties make it an ideal agent for treating central nervous system malignancies. In patients with advanced metastatic melanoma, brain metastases are a major cause of treatment failure. In this setting, TMZ has been shown to be as effective as dacarbazine, with a similar safety profile. More importantly, there is evidence to suggest that TMZ treated patients have a lower incidence of central nervous system relapse compared with dacarbazine-treated patients. Therefore, TMZ is actively being investigated for the treatment and prevention of brain metastases in melanoma patients. TMZ may become an important part of treatment regimens for advanced metastatic melanoma. PMID- 10794804 TI - Oral granisetron for the prevention of acute late onset nausea and vomiting in patients treated with moderately emetogenic chemotherapy. AB - PURPOSE: To demonstrate the efficacy of oral granisetron 1 mg twice daily for the prevention of late onset nausea and vomiting after moderately emetogenic chemotherapy that includes cyclophosphamide, carboplatin, or doxorubicin. METHODS: Prior to chemotherapy, patients were stratified by gender and randomized to receive oral granisetron (1 mg tablet twice daily) or prochlorperazine (10 mg sustained release capsule twice daily). Study agents were administered 1 h prior to and 12 h after chemotherapy. Antiemetics were administered for seven consecutive days. Efficacy variables were assessed 48 and 72 h after administration of chemotherapy, and included no emesis, no nausea, no moderate or severe nausea, and no antiemetic rescue. Safety analysis included all patients who received medication. RESULTS: A total of 230 patients were included in the intent-to-treat analysis; 119 patients received granisetron and 111 patients received prochlorperazine. Females, and all patients combined, who received granisetron had significantly higher no-emesis rates at 48 h (p =.010 and p =.016, respectively) than patients who received prochlorperazine. No-nausea rates at 48 h were numerically higher for all patients combined and females who received granisetron rather than prochlorperazine. Response rates for no nausea or mild nausea were also numerically higher in females treated with granisetron, compared to prochlorperazine, at 48 h. Significantly more patients (p <.001) and females (p <.001) in the granisetron group than in the prochlorperazine group did not require rescue antiemetics at 48 h. At 72 h, efficacy results were comparable for granisetron and prochlorperazine. CONCLUSION: Oral granisetron is well tolerated and more effective than prochlorperazine in preventing nausea and vomiting for up to 48 h following treatment with moderately emetogenic chemotherapy. PMID- 10794806 TI - Communicating genetic risk: pros, cons, and counsel. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded The Kenneth B. Schwartz Center at MGH. The Schwartz Center is a non-profit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to the patient, support to caregivers, and encourages the healing process. The center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. This case is of a woman with a personal, and a strong family history of breast cancer, who considered genetic testing for mutations in the BRCA1 and BRCA2 genes. The details of the case have been altered to protect the patient's anonymity. The patient was very anxious and there was disagreement between her healthcare providers about the potential benefits of genetic testing. The discussion of the case focused on several controversial issues, particularly the ownership of genetic information, and who is responsible for disseminating information to the family members at risk. The difficulties in communicating risk, providing emotional support and coping with the continuing uncertainties about screening and intervention are reviewed with an overview of the molecular biology, inheritance, and epidemiology of the BRCA1 and BRCA2 genes. PMID- 10794807 TI - Development of GnRH antagonists for prostate cancer: new approaches to treatment. AB - Prostate cancer has become the most common cancer among American men and is second only to lung cancer as a cause of male cancer-related death. Several treatment options exist for different stages of prostate cancer including observation, prostatectomy, radiation therapy, chemotherapy, and hormone therapy. Hormone therapy has evolved from the use of estrogens to gonadotropin-releasing hormone (GnRH) agonists and recently, investigational GnRH antagonists. GnRH receptor agonists such as leuprolide, bruserelin and goserelin have been used for the treatment of prostate cancer. These agonists eventually cause the inhibition of lutenizing hormone production, which in turn causes a suppression of testosterone and dihydrotestosterone, on which continued growth of prostate cancer cells depend. Several comparative studies of leuprorelin administered as daily injections or monthly depot injections have been reported. Disease progression was prevented in more than 72% of men administered daily leuprorelin, and in 82% to 89% of those receiving monthly depots. Another synthetic GnRH analog, goserelin, has been studied in a similar population of men with daily injections producing partial responses in 60% to 80% of men with previously untreated prostate cancer. Abarelix, a peptide antagonist of GnRH receptor, is also being studied for the treatment of prostate cancer. The discovery and development of GnRH antagonists may provide an important advance for patients with prostate cancer. Clearly the studies described herein, as well as many others, outline an exciting era of research to define the optimal use of hormonal therapy in prostate cancer. PMID- 10794808 TI - The molecular perspective: cyclooxygenase-2. PMID- 10794810 TI - The oncologist news bulletin PMID- 10794809 TI - The fourth biennial cancer meeting in Monaco. PMID- 10794811 TI - Cisplatin plus vinorelbine for patients with advanced head and neck squamous cell carcinoma. PMID- 10794812 TI - Pharmacological characterization of the cloned neuropeptide Y y(6) receptor. AB - Neuropeptide Y has potent appetite stimulating effects which are mediated by hypothalamic receptors believed to be of the neuropeptide Y Y(1) and/or neuropeptide Y Y(5) subtype. In mice, the neuropeptide Y y(6) receptor is also expressed in the hypothalamus, suggesting that it too may function as a feeding receptor in this species. Several laboratories have studied the pharmacology of the neuropeptide Y y(6) receptor, but their results are not in agreement. Using neuropeptide Y and a variety of peptide analogs and small molecule antagonists, we have determined that the pharmacology of the cloned mouse neuropeptide Y y(6) receptor is distinct from that of the other known neuropeptide Y receptors. The rank order of binding affinity for the mouse neuropeptide Y y(6) receptor is [(Ile, Glu,Pro,Dpr,Tyr,Arg,Leu,Arg,Tyr-NH(2))(2)human peptide YY=human, rat neuropeptide Y=human, rat neuropeptide Y-(2-36)=human, rat [Leu(31), Pro(34)porcine (Cys(2))-neuropeptide Y-(1-4)-8-aminooctanoyl-(D-Cys(27)porcine [D Trp(32)rat pancreatic polypeptide=human pancreatic polypeptide. A similar rank order of potency is seen for inhibition of forskolin-stimulated cyclic AMP. The neuropeptide Y Y(5) receptor antagonist trans-naphthalene-1-sulfonic acid ?4-[4 amino-quinazolin-2-ylamino)-methyl]-cyclohexylmethy l?-amide hydrochloride (CGP 71683A) and the neuropeptide Y Y(1) receptor antagonist ((R)-N(2)-diphenylacetyl) N-[(4-hydroxyphenyl)methyl]-argininam ide) (BIBP3226) bind weakly to the neuropeptide Y y(6) receptor (K(i)10, 000 nM, respectively). Although the function of the neuropeptide Y y(6) receptor remains to be elucidated, its pharmacology is not consistent with a role in appetite regulation. PMID- 10794814 TI - Regulation of activator protein-1-DNA binding activity by opioid peptides in estrogen-sensitive cells of rat hypothalamus and uterus. AB - The present studies demonstrate, for the first time, that the binding of activator protein-1 (AP-1)-DNA in rat uterus and the estrogen-sensitive areas of the hypothalamus, as measured by electrophoretic mobility shift assay, is increased 2 h after intraperitoneal injection of [D Met(2),Pro(5)]enkephalinamide. The effect was prevented by the opiate antagonist naltrexone given 30 min before the administration of [D Met(2),Pro(5)]enkephalinamide, suggesting the involvement of opioid peptide receptors in the observed effects. The present findings support the role of opioid peptides in the regulation of transcription in estrogen-sensitive cells. PMID- 10794813 TI - Inhibition of the human intermediate conductance Ca(2+)-activated K(+) channel, hIK1, by volatile anesthetics. AB - Ca(2+)-activated K(+) channels (K(Ca)) regulate a wide variety of cellular functions by coupling intracellular Ca(2+) concentration to membrane potential. There are three major groups of K(Ca) classified by their unit conductances: large (BK), intermediate (IK), and small (SK) conductance of channels. BK channel is gated by combined influences of Ca(2+) and voltage, while IK and SK channels are gated solely by Ca(2+). Volatile anesthetics inhibit BK channel activity by interfering with the Ca(2+) gating mechanism. However, the effects of anesthetics on IK and SK channels are unknown. Using cloned IK and SK channels, hIK1 and hSK1 3, respectively, we found that the currents of hIK1 were inhibited rapidly and reversibly by volatile anesthetics, whereas those of SK channels were not affected. The IC(50) values of the volatile anesthetics, halothane, sevoflurane, enflurane, and isoflurane for hIK1 inhibition were 0.69, 0.42, 1.01 and 1.03 mM, respectively, and were in the clinically used concentration range. In contrast to BK channel, halothane inhibition of hIK1 currents was independent of Ca(2+) concentration, suggesting that Ca(2+) gating mechanism is not involved. These results demonstrate that volatile anesthetics, such as halothane, enflurane, isoflurane, and sevoflurane, affect BK, IK, and SK channels in distinct ways. PMID- 10794815 TI - Selective antagonism by naloxonazine of antinociception by Tyr-D-Arg-Phe-beta Ala, a novel dermorphin analogue with high affinity at mu-opioid receptors. AB - To examine the role of mu-opioid receptor subtypes, we assessed the antinociceptive effect of H-Tyr-D-Arg-Phe-beta-Ala-OH (TAPA), an analogue of dermorphin N-terminal peptide in mice, using the tail-flick test. Intracerebroventricularly (i.c.v.) or intrathecally (i.t.) injected TAPA produced potent antinociception with tail-flick as a thermal noxious stimulus. The selective mu(1)-opioid receptor antagonist, naloxonazine (35 mg/kg, s.c.), or the selective mu-opioid receptor antagonist, beta-funaltrexamine, 24 h before testing antagonized the antinociceptive effect of i.t. or i.c.v. TAPA on the response to noxious stimuli. Pretreatment with beta-funaltrexamine completely antagonized the antinociception by both i.c.v. and i.t. administered TAPA and [D-Ala(2), Me Phe(4), Gly(ol)(5)]enkephalin (DAMGO). Especially in the tail-flick test, pretreatment with naloxonazine produced a marked rightward displacement of the i.t. TAPA dose-response curve for antinociception. Though DAMGO is a highly selective mu-opioid receptor agonist, pretreatment with naloxonazine partially blocked the antinociceptive response to DAMGO after i.c.v., but not after i. t. injection. These results indicate that TAPA can act as a highly selective mu(1) opioid receptor agonist (notable naloxonazine-sensitive receptor agonist) at not only the supraspinal level, but also the spinal level. These data also reveal different antinociceptive mechanisms for DAMGO and for TAPA. PMID- 10794816 TI - An acute i.c.v. infusion of leptin has no effect on hypothalamic histamine and tele-methylhistamine contents in Wistar rats. AB - The actions of intracerebroventricularly (i.c.v.) infused leptin on food intake, body weight and hypothalamic contents of histamine and tele-methylhistamine, the main histamine metabolite in the mammalian brain, were studied in male Wistar rats. The effect of the histamine H(1) receptor blockade on leptin-induced anorexia was also examined. It was found that leptin at the dose of 10 microg i.c.v. reduced 24-h food intake by 48% as compared with the controls (P<0.01). This leptin dose reduced feeding during 2-4 consecutive days. In spite of the marked changes in food consumption and body weight gain, leptin did not alter the hypothalamic contents of histamine and tele-methylhistamine. Furthermore, the blockade of histamine H(1) receptors by mepyramine did not attenuate the effect of leptin on feeding and body weight. The findings indicate that centrally administered leptin suppresses feeding and promotes weight loss through mechanisms that do not require the direct participation of the brain histaminergic neuron system. PMID- 10794817 TI - Antinociceptive and other behavioral effects of the steroid SC17599 are mediated by the mu-opioid receptor. AB - The objective of the present investigation was to evaluate the behavioral effects of SC17599 (17alpha-acetoxy-6-dimethylaminomethyl-21-fluoro-3-ethoxypregna -3, 5 dien-20-one) in mice and to determine if these effects are selectively mediated by opioid receptors. Although less potent than morphine, SC17599 produced dose dependent antinociception in both the acetic acid-induced writhing and warm water tail-withdrawal assays. Pretreatment with the opioid antagonist naltrexone and the noncompetitive mu-opioid receptor-selective antagonist methocinnamox, but not the delta-opioid receptor-selective antagonist naltrindole or the kappa-opioid receptor-selective antagonist nor-binaltorphimine, antagonized the antinociceptive effects of both SC17599 and morphine. Similarly to morphine, administration of SC17599 induced the Straub tail response in a dose-dependent and naltrexone-sensitive manner. At the highest doses studied, unlike morphine, SC17599 did not alter locomotor activity. The steroid SC17599 is structurally a very unusually selective mu-opioid agonist that produces behavioral effects, which are similar, but not identical, to those of morphine. PMID- 10794818 TI - gamma-aminobutyric acid mimetic drugs differentially inhibit the dopaminergic response to cocaine. AB - Dopaminergic activity in the mesocorticolimbic system is associated with reinforcing properties of psychostimulant drugs. We previously demonstrated that increased gamma-aminobutyric acid (GABA)-ergic activity produced by gamma-vinyl GABA [D,L-4-amino-hex-5-enoic acid (Vigabatrin(R))], an irreversible inhibitor of GABA-transaminase, attenuated cocaine, nicotine, heroin, alcohol, and methamphetamine-induced increases in extracellular nucleus accumbens dopamine as well as behaviors associated with these biochemical changes. In the present study, using in vivo microdialysis techniques, we compared three different strategies to increase GABAergic activity in order to modulate cocaine-induced increase in extracellular dopamine. Our data demonstrate that the anticonvulsant 1-(2-(((diphenylmethylene)amino)oxy)ethyl)-1,2,5, 6-tetrahydro-3 pyridinecarboxylic acid hydrochloride (NNC-711), a GABA uptake inhibitor, dose and time dependently diminished increases in extracellular dopamine following acute cocaine challenge. Furthermore, we demonstrated that cyclized analogue of vigabatrin, a competitive reversible GABA-transaminase inhibitor, is a more potent inhibitor of cocaine-induced dopamine increase than vigabatrin. Our data suggest that in addition to irreversible inhibition of GABA transaminase, inhibition of GABA uptake represent another potentially effective, indirect strategy for the treatment of cocaine abuse. PMID- 10794819 TI - Neuroprotective effect of 8-OH-DPAT in global cerebral ischemia assessed by stereological cell counting. AB - The neuroprotective effect of the 5-HT(1A) receptor agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) was tested in a 2-vessel occlusion model in rats. The post-ischemic core temperature was carefully monitored for 24 h. After 7 days of survival, the viable CA1 neurons were counted in an 8-OH-DPAT (125 microg/kg/h) and vehicle-treated group using the optical fractionator method. The vehicle-treated ischemic rats had a median number of dorsal CA1 neurons of 49,900 whereas the 8-OH-DPAT-treated ischemic rats had a significant lower median number of dorsal CA1 neurons 105,200 (P=0. 018). 8-OH-DPAT significantly lowered the core temperature compared to the vehicle-treated group during the 24-h post ischemic period. Hypothermia is proposed as a possible explanation of the neuroprotective effect of 8-OH-DPAT. PMID- 10794820 TI - Effects of WAY 100635 on antipsychotic-induced catalepsy in 5-HT depleted animals: a role for tonic activation of 5-HT(1A) receptors. AB - We recently observed that the 5-hydroxytryptamine (5-HT)(1A) receptor antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)-cycloh exanecarboxamide (WAY 100635) enhanced antipsychotic-induced catalepsy, which we hypothesized to be due to a blockade of tonic 5-HT(1A) receptor activation. Here, we examined this hypothesis by studying the effects of WAY 100635 in animals that were depleted of 5-HT by repeated treatment with the 5-HT synthesis inhibitor p chlorophenylalanine methyl ester. Depletion of 5-HT abolished the enhancement by WAY 100635 of catalepsy induced by low doses of the antipsychotics nemonapride and raclopride, in agreement with the hypothesis that WAY 100635 enhances catalepsy by blocking tonic 5-HT(1A) receptor activation. Given the predictive validity of catalepsy, these findings indicate that 5-HT(1A) receptor blockade may enhance the extrapyramidal side-effects of antipsychotics in humans. PMID- 10794822 TI - Chronic lithium chloride fails to prevent imipramine-induced sensitization to the dopamine D(2)-like receptor agonist quinpirole. AB - Lithium salts, an effective antimanic treatment, are able to prevent the development of the dopaminergic behavioural supersensitivity induced by chronic treatment with neuroleptics, by denervation of the dopaminergic terminal fields and by rapid eye movements (REM) sleep deprivation, which is considered a model of mania. We have studied the effect of a lithium (LiCl) diet, inducing a lithium serum level in the range of therapeutic efficacy, on the development of the supersensitivity to the locomotor effect of the dopamine D(2)-like receptor agonist, quinpirole, induced by chronic treatment with the antidepressant drug, imipramine. The results show that lithium is not able to prevent the development of such behavioural supersensitivity. The present data suggest that antidepressant-induced dopaminergic supersensitivity might provide a useful model of those manic states induced by (or subsequent to) antidepressant treatments. Moreover, the finding is consistent with the view that antidepressant-induced dopaminergic supersensitivity might play a role in the therapeutic effect of these drugs (which is known to be augmented by lithium, and not antagonised). Finally, the results show that the dopaminergic supersensitivity induced by imipramine is qualitatively different from that induced by neuroleptics or denervation of the dopaminergic terminal fields. PMID- 10794821 TI - Improvement by low doses of nociceptin on scopolamine-induced impairment of learning and/or memory. AB - The effects of fmol doses of nociceptin/orphanin FQ on scopolamine-induced impairment of learning and/or memory were examined using spontaneous alternation of Y-maze and step-down type passive avoidance tasks. While fmol doses of nociceptin alone had no effect on spontaneous alternation or passive avoidance behavior in normal mice, administration of nociceptin (10 and/or 100 fmol/mouse) 30 min before spontaneous alternation performance or the training session of the passive avoidance task, significantly improved the scopolamine-induced impairment of spontaneous alternation and passive avoidance behavior. This ameliorating effect was not antagonized by nocistatin (0.5 and 5.0 nmol/mouse, i.c.v.), naloxone benzoylhydrazone (2.3, 11.2, and 56.1 micromol/kg, s.c.) or nor binaltorphimine (4.9 nmol/mouse, i.c.v.). These results indicated that very low doses of nociceptin ameliorate impairments of spontaneous alternation and passive avoidance induced by scopolamine, and suggested that this peptide has bidirectional modulatory effects on learning and memory; impairment at high doses and amelioration at low doses. PMID- 10794823 TI - Involvement of ATP-sensitive K(+) channels in the anti-tussive effect of moguisteine. AB - The effect of glibenclamide, an ATP-sensitive K(+) channel blocker, on the anti tussive effect of moguisteine and of pinacidil, an ATP-sensitive K(+) channel opener in guinea pigs was studied. Pinacidil (1 and 5 mg/kg, subcutaneous (s.c.)) dose-dependently reduced the number of coughs. The anti-tussive effect of pinacidil was significantly and dose-dependently antagonized by pre-treatment with glibenclamide (3 and 10 mg/kg, i.p.). Moguisteine (1 and 5 mg/kg, s.c.) dose dependently reduced the number of coughs. The anti-tussive effect of moguisteine was also reduced by pre-treatment with glibenclamide, in a dose-dependent manner. However, pre-treatment with glibenclamide had no effect on the anti-tussive effects of dihydrocodeine and dextromethorphan. Glibenclamide (10 mg/kg, i.p.), by itself, had no significant effect on the number of coughs. These results suggest that pinacidil and moguisteine may exert their anti-tussive effects through the activation of ATP-sensitive K(+) channels. Furthermore, it is possible that ATP-sensitive K(+) channels may be involved in the anti-tussive effect of peripherally acting non-narcotic anti-tussive drugs. PMID- 10794825 TI - Multixenobiotic resistance as a cellular defense mechanism in aquatic organisms. AB - Multixenobiotic resistance in aquatic organisms exposed to natural toxins or anthropogenic contaminants is a phenomenon analogous to multidrug resistance in mammalian tumor cell lines tolerant of anti-cancer drugs. Multidrug resistance is commonly due to the elevated expression of transmembrane P-glycoproteins (P-gp) which actively transport a wide variety of structurally and functionally diverse compounds. The purpose of this review is to place aquatic ecotoxicological data in context of the larger multidrug resistance field of study. Information on P glycoproteins structure, mechanism of transport, and substrate specificity gained through traditional mammalian and cell culture models is examined in conjunction with recent work on aquatic species exposed to xenobiotics both in the field and in the laboratory. The physiological function of P-glycoproteins is explored through studies of gene knockout models and expression patterns in normal tissues and tumors. The effect of xenobiotic exposures on P-gp activity and protein titer is examined in wild and captive populations of aquatic invertebrates and vertebrates. Substrate overlap and evidence of co-expression of phase I detoxification enzymes (e.g. cytochromes P450) and P-gp are presented. The role of P-gp chemosensitizers as environmental pollutants and the ecotoxicological consequences of P-gp inhibition are highlighted. The overwhelming evidence suggests that P-glycoproteins provide aquatic organisms with resistance to a wide range of natural and anthropogenic toxins. PMID- 10794824 TI - EM574, an erythromycin derivative, improves delayed gastric emptying of semi solid meals in conscious dogs. AB - The gastroprokinetic effects of de(N-methyl)-N-isopropyl-8, 9-anhydroerythromycin A 6,9-hemiacetal (EM574), a non-peptide motilin receptor agonist, were investigated in conscious dogs in a normal state and with experimentally-induced gastroparesis. Gastric emptying of semi-solid meals was assessed indirectly from acetaminophen absorption with simultaneous recording of gastric antral motility. In the normal state, post-prandial intraduodenal administration of EM574 (0.03 mg/kg) [corrected] stimulated antral motility and significantly enhanced gastric emptying as potently as did intravenous porcine motilin (0.003 mg/kg/h). Intraduodenal cisapride at 1 mg/kg denal cisapride at 1 mg/kg elicited antral contractions and tended to accelerate gastric emptying but at 3 mg/kg, gastric emptying was not enhanced despite a further increase in the motor index. In dogs with gastroparesis induced by intraduodenal oleic acid or intravenous dopamine, EM574 (0.03 mg/kg) increased antral motility and reversed the delayed gastric emptying completely. Cisapride (1 mg/kg) partially ameliorated the impaired emptying under these conditions. In atropinized dogs, no acceleration of gastric emptying by EM574 was observed. These results indicate that EM574 potently accelerates gastric emptying of caloric meals in dogs in a normal state and with experimentally-induced gastroparesis, and also suggest that the effect is mediated through stimulation of a cholinergic neural pathway. PMID- 10794826 TI - Hepatic activities of thiamine-dependent enzymes, glucose-6-phosphate dehydrogenase and cytochrome P4501A in Baltic salmon (Salmo salar) yolk-sac fry after thiamine treatment. AB - Sea-run Baltic salmon (Salmo salar) populations have been affected by the M74 syndrome since 1974 causing high yolk-sac fry losses in Swedish compensatory rearing plants. M74 has been shown to be a maternally transmitted thiamine (vitamin B(1)) deficiency. The aim of this study was to investigate possible relationships between thiamine and hepatic activities of the thiamine-dependent enzymes transketolase (TK) and alpha-ketoglutarate dehydrogenase (alpha-KGDH) in addition to glucose-6-phosphate dehydrogenase (G6PDH) and cytochrome P4501A (CYP1A), measured as 7-ethoxyresorufin O-deethylase (EROD), in Baltic salmon yolk sac fry after treatment with thiamine. Thiamine concentrations and activities of TK, alpha-KGDH and EROD were significantly lower (P<0.05) in M74 groups compared to controls (not developing M74) and family groups of thiamine injected females. In M74-developing groups the thiamine immersions reduced the mortality from 86 to 13% and restored thiamine concentrations and activities of TK, alpha-KGDH and EROD to levels slightly lower than the immersed controls. An interesting fact was that the controls showed significantly elevated (P<0.05) TK and alpha-KGDH activities after immersions in thiamine, indicating that they also may have a stressed thiamine metabolism. The TK and alpha-KGDH-activities of unimmersed groups correlated significantly (P<0.05) with the thiamine content. We suggest that the low activities of TK and alpha-KGDH in M74 groups may be an integrative part in the pathogenesis of M74 development. PMID- 10794827 TI - The role of development and duration of exposure to the embryotoxicity of diazinon. AB - Medaka, Oryzias latipes, were used as a laboratory surrogate for species of concern to define the effects of diazinon exposure on teleost embryogenesis. Medaka embryos were placed in a static, non-renewal system and exposures initiated on days 1, 3, or 5 of development. Following initiation of exposure, replicates (n=5) remained in diazinon for a total of 4 days or from the day of initiation to day 9 of development. This exposure scenario was designed to elucidate sensitive periods in development for diazinon-induced toxicity but also shows the effect of added exposure duration on the degree of toxicity. Embryos were observed daily and endpoints recorded included: edema formation, total hatch, mean day of hatch, percentage of larvae with swim bladder inflation, and total length of larvae on day 14, when observations were terminated. Diazinon exposure resulted in decreases in hatch success, swim bladder inflation and the total length of larvae. In addition, dose-response increases in the incidence of edemas of the pericardial sac and vitelline veins were recorded. As expected, severity of embryotoxicity was positively correlated with duration of exposure. While no developmental period was the most sensitive for all toxic effects, for certain endpoints the severity of effects was dependent on exposure timing. Total hatch was greatly affected in embryos exposed from day 1 until day 5 whereas edema was more prevalent in embryos exposed later in development. Finally, among endpoints recorded, total length of larvae was the most sensitive indicator of exposure with all exposure groups showing significant (P<0.05) decreases in length at 5 ppm. PMID- 10794828 TI - Tributyltin induces cytoskeletal alterations in the colonial ascidian Botryllus schlosseri phagocytes via interaction with calmodulin. AB - In the colonial ascidian Botryllus schlosseri, tributyltin (TBT), a powerful antifouling biocide, acts as immunotoxic xenobiotic since, at a sublethal concentration (10 uM), it causes an irreversible and significant decrease in in vitro yeast phagocytosis, associated with considerable changes in the shape of phagocytes, which withdraw their pseudopodia and become spherical, due to structural damage of cytoskeletal components. The addition of TBT to the culture medium causes a significant decrease in the amoebocytic index, i.e. the percentage of amoeboid-shaped haemocytes, and prolonged washing in sea water never succeeds in restoring amoeboid shape. In these cytoskeletal alterations, F actin undergoes extensive depolymerisation, resulting in the absence of FITC phalloidin fluorescence. Microtubules are not recognisable as single filaments with anti-alpha-tubulin immunofluorescence, although the centrosome is not affected. The addition of increasing exogenous calmodulin (CaM) concentrations (from 20 to 120 uM) after incubation in TBT determines a significant increase in the amoebocytic index, although it is not able to bring it to that of controls, suggesting that CaM in the medium in any case externally exerts an influence on haemocytes pretreated with TBT. The copresence of TBT and exogenous CaM at concentrations higher than 80 ug/ml restores the amoebocytic index and cytoskeletal morphology. The latter appears complete for microtubules and partial for microfilaments. Experiments with isodynamic mixtures of TBT and specific CaM inhibitors, i.e. chlorpromazine (CPZ) and N-(6-aminohexyl)-5-chloronaphtalene-1 sulfonamide (W-7), reveal the synergistic effect of antagonism, indicating competition for the same site - a Ca(2+)-CaM hydrophobic region - by both interacting substances and, therefore, the formation of a TBT-CaM complex. Instead, isodynamic mixtures with thapsigargin, an inhibitor of Ca(2+)-ATPase of the endoplasmic reticulum, have an effect of potentiation, suggesting that TBT indirectly interacts with this Ca(2+)-ATPase activity. We hypothesise that the main mechanism of action of TBT in B. schlosseri phagocytes is alteration of Ca(2+) homeostasis by means of direct interaction with endogenous CaM, which induces a conformational change preventing the regulative activity of CaM on Ca(2+)-ATPase. Consequently, an excess of cytosolic Ca(2+) accumulates which, together with the inhibition of CaM-dependent kinases and Ca(2+)-regulated proteins, produces extensive cytoskeletal disorganisation. PMID- 10794829 TI - Biochemical indicators of pollution exposure in shorthorn sculpin (Myoxocephalus scorpius), caught in four harbours on the southwest coast of Iceland. AB - Shorthorn sculpins (Myoxocephalus scorpius) were caught in four Icelandic harbours, differing in size, use and traffic. Biochemical responses in liver were measured and chemicals analysed in bile. Eyrarbakki harbour, which has not been in use for many years was chosen as a control site. Njar partial differentialvik harbour is a small fishing harbour and a marina, Sandger partial differentiali harbour is a large fishing harbour, and Reykjavik harbour is a large fishing harbour and an international transport harbour. Higher levels of DNA-adducts and cytochrome P4501A (CYP1A) in the fish from the harbours in Sandger partial differentiali, Njar partial differentialvik and Reykjavik, compared to Eyrarbakki harbour, indicate PAH exposure. This was confirmed by PAH analysis in bile. The higher activities of the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in fish caught in Sandger partial differentiali, than in fish caught in the other harbours, indicate exposure of sculpin to prooxidative compounds in Sandger partial differentiali harbour. Shorthorn sculpin seems to be a convenient species for monitoring pollution in northern coastal areas. PMID- 10794830 TI - Organochlorine residues in european eel (Anguilla anguilla), crucian carp (Carassius carassius) and catfish (Ictalurus nebulosus) from Vaccares lagoon (French National Nature Reserve of Camargue) - effects on some physiological parameters. AB - European eels (Anguilla anguilla), crucian carps (Carassius carassius) and catfish (Ictalurus nebulosus) were collected in three coastal locations of the Vaccares lagoon (French National Nature Reserve of Camargue). The purpose of this paper is to report results of the first biomonitoring investigation in fish living in this protected coastal wetland. Residues of organochlorine (OC) contaminants (i.e. SigmaPCBs, gammaHCH, HCB, dieldrin, pp'-DDE) were determined in hepatic and muscular tissues, in order to compare geographical and seasonal distribution. Total exposure levels appeared to be more important in fatty fish such as eels than in crucian carps and catfish. The highest OC concentrations in liver (SigmaPCB) and in muscle (gammaHCH) were detected in Spring in some fish coming from a site located near the mouth of a canal draining irrigation waters of rice fields. Morphophysiological parameters (condition factor, organo-somatic indexes and lipid tissue composition) were measured concomitantly. Correlations between the hepatic and muscular burdens of OC and condition factor or organo somatic indexes were infrequent and rather negative. Localization of lipid accumulation (neutral or polar lipids) depended on metabolic rates of different species and appeared related to the fish trophic level. PMID- 10794831 TI - Effects of the mammalian antiandrogen vinclozolin on development and reproduction of the fathead minnow (Pimephales promelas). AB - Previous work with the chlorinated fungicide vinclozolin and its metabolites, 2 {[(3,5-dichlorophenyl)-carbamoyl]oxy}-2-methyl-3-butenoic acid (M1) and 3',5' dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), indicated antiandrogenic properties expressed in vivo as abnormalities in sexual differentiation of male rats after maternal exposures. In this study, we attempted to determine whether vinclozolin might also exhibit antiandrogenic properties in a model fish species, the fathead minnow, Pimephales promelas. In one study, embryonic (<6 h old) fathead minnows were exposed for approximately 34 days to five toxicant concentrations, ranging from 90 to 1200 ug l(-1), delivered via a flow-through diluter. The embryos were periodically sampled to determine survival, growth and gross pathology, and then placed in clean water for 4-6 months to assess long term effects on sexual differentiation and subsequent reproductive success. Except for slightly reduced growth after 34 days in the highest vinclozolin concentration, no adverse effects were noted with respect to any of these endpoints. In a second experiment, adult fathead minnows were exposed to vinclozolin concentrations of approximately 200 or 700 ug l(-1) for 21 days, following which, gonadal morphology was assessed and serum sex steroid concentrations determined. Tissue samples from the exposed adults were assayed for vinclozolin and its metabolites. There was a slight increase in the serum beta-estradiol concentration of the male fathead minnows exposed to 700 ug vinclozolin l(-1), and a marked reduction in gonadal condition of female fish from this treatment. The possibility that vinclozolin and its metabolites would bind to androgen receptors in the fathead minnow was investigated through competitive radioligand binding studies. Vinclozolin, M1 and M2 failed to compete for high-affinity, low-capacity testosterone binding sites in fathead minnow brain and ovary cytosolic fractions, suggesting that these chemicals might not act as antiandrogens in the fathead minnow. More experimentation is necessary to determine whether responses observed in vivo might be due to the effects of vinclozolin (or its metabolites) on some other aspect of endocrine function. PMID- 10794832 TI - Accumulation and distribution of dietary uranium in lake whitefish (Coregonus clupeaformis). AB - We investigated the accumulation and distribution of uranium (U) in adult lake whitefish (Coregonus clupeaformis) fed a commercial diet containing 100, 1000 and 10000 ug U/g for 10, 30, and 100 days. No food avoidance or refusal occurred. The major sites of U accumulation were in the mineralized tissues, bone and scales, and in intestine, liver, kidney, and, in the highest treatment, gonads. Significant accumulation in fish fed 100 ug U/g was observed only in scales. Duration-dependent accumulation was observed in bone, scales, liver, and kidney of fish fed 10000 ug U/g and in scales of fish fed 1000 ug U/g. Dose-dependent accumulation was observed in scales of fish exposed for 100 days. U accumulation in gonads and gill peaked on day 30 when fish gonads were in the most advanced stage of maturation, of the sampling days evaluated. To evaluate the biological availability of U to fish inhabiting contaminated aquatic systems, analyses of U in scales, bone, intestine, kidney, and liver are recommended for biomonitoring programmes. The toxicology of U in these fish is described in the following manuscript (Cooley, H.M., Evans, R.E. Klaverkamp, J.F., 2000. Toxicology of dietary uranium in lake whitefish (coregonus clupeaformis) Aquat. Toxicol., 48, 393-413). PMID- 10794833 TI - Toxicology of dietary uranium in lake whitefish (Coregonus clupeaformis). AB - Using responses ranging from whole animal to the molecular level, the toxicological significance of the accumulation and distribution of uranium (U) in adult lake whitefish fed a commercial diet contaminated with three concentrations of U, 100, 1000 and 10000 ug U/g, for 10, 30, and 100 days, as described in the previous manuscript (Cooley, H.M., Klaverkamp, J.F., 2000. Accumulation and distribution of dietary uranium in Lake whitefish (Coregonus clupeaformis), Aquat. Toxicol., 48, 375-392), was evaluated. Whole organism morphometrics were unaltered by U exposure. Haematological variables were either unchanged or only transiently affected. Liver and kidney metallothionein concentrations were largely unaltered. Concentrations of serum lipid peroxides were significantly elevated in all treatment groups on days 30 and 100, indicating that U may damage cellular and sub-cellular membranes. Dose- and duration-dependent histopathologies were present in liver and posterior kidney of treated lake whitefish. The most consistent and pronounced lesions in liver were focal hepatocyte necrosis and alterations of bile ductule epithelium. Dose- and duration-dependent renal lesions were most evident in proximal tubules. However, we observed a range of histopathologies in this organ, including tubular necrosis, inflammation, haemorrhaging, depletion of haematopoietic tissues, alterations of distal tubules and collecting ducts, tubule dilation, pigmented macrophage proliferation, and glomerular lesions. Pathologies were observed in lake whitefish at all concentrations of dietary U following prolonged exposure. When used in conjunction with U tissue residue analyses, histopathological lesions and indices of lipid peroxidation may prove useful indicators of U toxicity when assessing fish health in U biomonitoring programmes. PMID- 10794834 TI - Morphological and biochemical changes in the gills of Tilapia (Oreochromis mossambicus) to ambient cadmium exposure. AB - Tilapia, Oreochromis mossambicus, were reared in freshwater and exposed to 40, 80 and 160 ppb ambient cadmium for a period of 7 days to investigate the effects of short-term Cd exposure on the cellular function of gill chloride cells. Gills were sampled after 3 and 7 days exposure. The accumulated Cd concentration in gills was analyzed using a graphite furnace atomic absorption spectrophotometer while the morphological changes of pavement and chloride cells were examined using scanning and transmission electron microscopes. Gill Cd concentration was significantly increased in samples (10.03-44.36 ppb) at 3 and 7 days exposure. Accompanying this was an augmentation of microridge in pavement cells, and an increase in the density (1964-3603 /mm(2)) and apical membrane area (11.57-46.32 um(2)) in chloride cells, indicating an adaptational modification of the cell morphology in assisting gaseous transfer and Ca(2+) uptake, respectively. However, biochemical analyses of the gill tissues enumerated a decrease in both the activities of alkaline and high-affinity Ca(2+)-ATPases. This indicated a reduction in the Ca(2+)-transport capacity per unit chloride cell, suggesting chloride cells being the primary target of Cd which subsequently lead to fish hypocalcemia. PMID- 10794835 TI - Cytochrome P450 enzymes in aquatic invertebrates: recent advances and future directions. AB - A variety of enzymes and other proteins are produced by organisms in response to xenobiotic exposures. Cytochrome P450s (CYP) are one of the major phase I-type classes of detoxification enzymes found in terrestrial and aquatic organisms ranging from bacteria to vertebrates. These enzymes metabolize a wide variety of substrates including endogenous molecules (e.g. fatty acids, eicosenoids, steroids) and xenobiotics (e.g. hydrocarbons, pesticides, drugs). Aquatic invertebrates, especially those in marine habitats, occupy every aspect of the environment, from above the surface (intertidal) to below the sediments. In turn, they have extremely diverse physiologies and are exposed to a vast array of potential toxicants. Aspects of aquatic invertebrate cytochrome P450 enzymes have been studied for the last 25 years. In a few phyla, P450 activities have been measured and are responsive to xenobiotic exposures. Until the last several years, little progress had occurred in the identification of P450 gene diversity in aquatic invertebrates. Molecular biology tools have greatly aided this search, and are likely to identify as much diversity for this protein superfamily as is present in higher marine and terrestrial organisms. Recent work has expanded our knowledge of the CYP superfamily, and new developments will rapidly advance the usefulness of these genes into such fields as biomarker research. Advances of the last decade are reviewed and insights are presented from related insect studies. PMID- 10794836 TI - Polycyclic aromatic hydrocarbons inhibit in vitro ovarian steroidogenesis in the flounder (Platichthys flesus L.). AB - The in vitro effect of polycyclic aromatic hydrocarbons (PAHs) on ovarian steroidogenesis of the flounder (Platichthys flesus L.) was determined. Fully vitellogenic ovary tissue was in vitro incubated in the presence of phenanthrene, benzo[a]pyrene or chrysene, using 17alpha-hydroxyprogesterone or androstenedione as precursors. Androstenedione (A), testosterone (T) and 17beta-estradiol (E(2)) synthesised in the presence of PAHs were assayed by radioimmunoassay and results compared with control incubations. In order to establish the effect of PAHs on the steroidogenic enzyme systems cytochrome P450 17,20-lyase (P450-17,20l), 17beta-hydroxysteroid dehydrogenase (17beta-HSD) and cytochrome P450 aromatase (P450-arom), results were also compared with the action of ketoconazole (KCZ) and aminoglutethimide (AMG), wich are, respectively, inhibitors of cytochrome P450 steroidogenic enzymes and of P450-arom. KCZ inhibited secretion of A and E(2) in 65% and T in 40%, as a consequence of inhibited P450-17,20l and P450-arom. AMG inhibited P450-arom, which resulted in decreased E(2) synthesis to approximately 50% of control incubations. All the three PAHs inhibited A secretion by approximately 50% and E(2) from 10 to 40%. Because steroid conjugation was also inhibited by phenanthrene, it could be concluded that PAH action was mediated by an inhibitory effect over P450-17,20l, 17beta-HSD and P450-arom. Except for 17beta-HSD, PAHs resembled KCZ, and P450-17,20l was the most sensitive to their inhibitory effect. In conclusion, PAHs strongly blocked the activity of P450 17,20l, a rate-limiting enzyme for conversion of C21 to C19 steroids, and showed, therefore, the potential to disrupt the reproductive cycle of fish living in polluted environments, due to impairment of steroid biosynthesis. PMID- 10794837 TI - Inhibition of eel enzymatic activities by cadmium. AB - The aim of the present work was to study the in vitro effect of cadmium on enzymes, such as intestinal and branchial carbonic anhydrase (CA) and Na(+)-K(+) ATPase which play a key role in salt- and osmoregulation and acid-base balance in the teleost fish, Anguilla anguilla. Carbonic anhydrase activities in gill and intestinal homogenates were significantly inhibited by CdCl(2), the gill CA being more sensitive to the heavy metal (IC(50) for the branchial CA=9.97+/-1.03x10(-6) M, IC(50) for the intestinal CA=3.64+/-1.03x10(-5) M, P<0.01). With regards to the intestinal CA activity, it has been shown in a previous study (Maffia et al., 1996) that two isoforms exist, a cytosolic and a brush-border membrane bound. These two isoforms show a different sensitivity to cadmium, with the membrane bound enzyme less sensitive with respect to the cytosolic one, since it showed still an incomplete inhibition at the highest cadmium concentration tested. The inhibition of all the CA activity tested revealed a time-dependence since it required at least 10 min (1 h for the membrane-bound isoform) preincubation with the heavy metal to appear. Na(+)-K(+)-ATPase enzymatic activities, measured in intestinal and branchial homogenates, were inhibited by cadmium in a dose dependent manner, with the branchial activity being more sensitive to the action of the heavy metal than the intestinal one (IC(50) for the branchial enzyme=1.38+/-0.09x10(-7) M, IC(50) for the intestinal enzyme=2.86+/-0.02x10(-7) M, P<0.01). The most of inhibition of the enzyme appeared without any preincubation with the heavy metal. Mg(2+)-ATPase activity was not significantly altered by the in vitro cadmium exposure either in the gills or in the intestine. These findings observed in vitro could be useful in the understanding of the toxic effects that cadmium elicits on aquatic organisms in vivo. In fact, the impairment of the activity of enzymes which carry out key physiological roles could cause alterations of the physiology of the whole organism. PMID- 10794838 TI - The role of the primary care physician in the Israeli health care system as a 'gatekeeper'--the viewpoint of health care policy makers. AB - The aim of the study was to determine the attitudes of policy makers in the health care system in Israel to a change in the role of primary care physicians (PCP) and to ascertain the conditions under which they would be ready to adopt the model of PCP as gatekeeper. The study design was qualitative, with analyses of in-depth structured interviews of 20 policy makers from the Ministry of Health, the Sick Funds' central administrations and the Israel Medical Association (IMA) central office. The majority of the respondents claim that they want highly trained PCPs (family physicians, pediatricians and internals) to play a central role in the health care system. They should be co-ordinators, highly accessible and should be able to weigh cost considerations. However, only about half of the respondents support a full gatekeeper model and most of them think that the gatekeeper concept has a negative connotation. They also feel that it would be difficult to implement regulations regarding primary care. The barriers to implementation of the gatekeeper model, as cited by the respondents include loss of faith in PCPs by the general population, dearth of PCPs with adequate training, low stature, lack of availability on a 24-h basis, resistance by specialists, strong competition between the sick funds including promises of direct access to specialists, the medical care habits of the general population many of whom do not settle for only one opinion, and a declared anti-gatekeeper policy by one of the sick funds. Ways to overcome these obstacles include implementation of fundholding clinics, patient education on the importance of having a personal physician, appropriate marketing by family medicine and primary care advocates, and continued training in primary care. Israeli health care policy makers have an ambivalent attitude to strengthening the role of primary care. In theory, they profess support for placing primary care physicians in a central role in the health care system. However, in practice almost half oppose the full gatekeeper model. Therefore, introduction of a gatekeeper model into the Israeli health care system should be implemented gradually, based on incentives rather than regulations. Furthermore, the idea should be marketed by the primary care physicians' professional organizations, the Ministry of Health and the sick funds to physicians as well as to patients, in order to garner their support. In light of the broad consensus that competent primary care physicians are the basis for implementation of the gatekeeper model, board certification should be gradually required by employers of primary care physicians. The process of training physicians currently working in the system should be encouraged and supported by the Ministry of Health. Given the existing opposition of policy makers to giving primary care physicians exclusive referral rights to specialists, the current policy of direct access to a limited number of specialties should be continued but not extended to other specialties. PMID- 10794839 TI - The determinants of the public-private mix in Canadian health care expenditures: 1975-1996. AB - The health care policy issue regarding the balance between public and private health spending is examined. An empirical model of the determinants of the public private mix in Canadian health care expenditures over the period 1975-1996 is estimated for total health care expenditures as well as separate expenditure categories such as hospitals, physicians and drugs. The results find that the key determinants of the split are per capita income, government transfer variables and the share of individual income held by the top quintile of the income distribution. Much of the public-private split is determined by long term economic forces. However, the importance of the federal health transfer variables and the variables representing shifts in fiscal transfer regimes suggest the increase in the private share of health spending since 1975 is also partly the result of the policy choice to reduce federal health transfers. PMID- 10794840 TI - Decentralising the health sector: issues in Brazil. AB - The health sector in Brazil has undergone important changes, particularly with the development of the Unified Health System (SUS). Decentralisation is an important principle of SUS and advances have been made in transferring responsibilities and resources to the local government units, known as municipios. This article describes the changes introduced, focusing on the system of municipio classification and the funding mechanisms introduced through the basic operating rule (BOR) of 1996. The paper then moves on to analysing three key issues of decentralisation in Brazil that are related to the policy process, the system of decentralisation and the output of decentralisation. Firstly, the formal process by which decisions on health sector reform are made is discussed with particular attention being paid to the negotiated and relatively open policy space. Secondly, the role of the states is discussed within the decentralised system. Thirdly, the impact of decentralisation on equity is discussed with particular reference to the resourcing of the Municipal Health Funds. The article concludes by emphasising the political nature of health sector decentralisation and the need to develop the conditions for effectiveness in decentralisation programmes. PMID- 10794841 TI - The cost of prescription medicines to patients. AB - The study compares the cost-sharing (co-payment) arrangements for prescribed medicines in a sample of EU countries. Through a set of typical prescription scenarios, the cost burden to individual patients of prescriptions are examined, in the context of drug price, and from the perspective of therapeutic need. The cost to patients of medicines is consistently lower in some, and higher in other, countries, regardless of the type of prescription charge system. Fixed charge systems, as opposed to graduated co-payment systems, are obviously more likely to lead to similar charges for the treatment of comparable clinical conditions, but depending on the level of the charge, can result in the patient paying a higher charge than the price of the drug to the health organisation. Exemption from charges for prescription medicines, commonly relate to clinical condition and level of income. Some systems also have age-related criteria and apply ceilings to the total prescription cost burden borne by the patient. The impact on patient costs of specific policy formulations is discussed and a proposal is made for cost convergence for comparable therapies. The method used in this study may also provide a route for investigating model systems prior to implementation. PMID- 10794842 TI - Formation of diffuse and fibrillar tangles in aging and early Alzheimer's disease. AB - The changes in tau that are associated with the early formation of tangles in aging and in preclinical and very mild Alzheimer's Disease (AD) were studied with two antibodies against AD-specific tau: PHF-1, which recognizes a phosphorylated epitope at Ser396 through 404, and MC-1, which recognizes a folded, conformational epitope that includes amino acids at both 7 through 9 and 312 through 342. Both antibodies demonstrated cells with diffuse or granular staining (diffuse tangles) and cells with fibrillar staining (fibrillar tangles). The fibrillar tangles corresponded to classical tangles and increase exponentially with age and severity of AD. The diffuse tangles seemed to represent an earlier form of tangles; their density peaked around preclinical AD, and then decreased in more severe stages of AD. MC-1 consistently stained more diffuse tangles than PHF-1, suggesting that the conformational change in tau precedes phosphorylation at the PHF-1 epitope during paired helical filament formation. PMID- 10794843 TI - Choline acetyltransferase activity and cognitive domain scores of Alzheimer's patients. AB - Choline acetyltransferase activity and cognitive domain scores of Alzheimer's patients. Item scores from the Mattis Dementia Rating Scale (MDRS) and the Mini Mental State Examination (MMSE) from 389 patients with probable Alzheimer's disease were submitted to principal component analysis with orthogonal rotation. The optimal solution identified four factors that reflected the cognitive domains of attention/registration, verbal fluency/reasoning, graphomotor/praxis and recent memory. A subgroup of patients was identified for whom both the MDRS and the MMSE had been administered within the 12 months before death. Scores were assigned to these patients for the four factors. These cognitive-domain scores were then correlated with postmortem choline acetyltransferase (ChAT) activity in the medial frontal cortex, inferior parietal cortex, and hippocampus. ChAT activity in both the medial frontal and the inferior parietal cortex significantly correlated with scores on the graphomotor/praxis factor. Medial frontal ChAT also correlated significantly with the attention/registration scores. Hippocampal ChAT correlated significantly only with recent memory scores. These results are consistent with current animal research regarding the effect of selective cholinergic lesions on behavior. PMID- 10794844 TI - Atrophy of the medial occipitotemporal, inferior, and middle temporal gyri in non demented elderly predict decline to Alzheimer's disease. AB - Our goal was to ascertain, among normal elderly and individuals with mild cognitive impairment, which temporal lobe neocortical regions predicted decline to dementia of the Alzheimer's type (DAT). Individuals received an MRI at baseline and a clinical and cognitive evaluation at baseline and follow-up. By using the baseline MRI we assessed the anatomical subdivisions of the temporal lobe: anteromedial temporal lobe (hippocampus and parahippocampal gyrus), medial occipitotemporal (fusiform) gyrus, middle and inferior temporal gyri, and superior temporal gyrus. We studied two groups of carefully screened age- and education-matched elderly individuals: 26 normal elderly (NL) and 20 individuals with mild cognitive impairment (MCI). Fourteen individuals (12 from the MCI group and two from the NL group) declined to DAT within the 3.2-year follow-up interval. We used logistic regression analyses to ascertain whether the baseline brain volumes were useful predictors of decline to DAT at follow-up after accounting for age, gender, individual differences in brain size, and other variables known to predict DAT. After accounting for age, gender, and head size, adding the volume of the anteromedial temporal lobe (the aggregate of hippocampus and parahippocampal gyrus) and an index of global atrophy raised the accuracy of overall classification to 80.4%. However, the ability to detect those individuals who declined (sensitivity) was low at 57%. When baseline medial occipitotemporal and the combined middle and inferior temporal gyri were added to the logistic model, the overall classification accuracy reached 95.6% and, most importantly, the sensitivity rose to 92.8%. These data indicate that the medial occipitotemporal and the combined middle and inferior temporal gyri may be the first temporal lobe neocortical sites affected in AD; atrophy in these areas may herald the presence of future AD among nondemented individuals. No other clinical baseline variables examined predicted decline with sensitivities above 71%. The apolipoprotein APOE epsilon4 genotype was not associated with decline. PMID- 10794845 TI - Decreased high-density lipoprotein cholesterol and serum apolipoprotein AI concentrations are highly correlated with the severity of Alzheimer's disease. AB - Serum apolipoprotein (apo) AI concentration was studied in 98 Alzheimer's disease (AD) patients (77.56+/-8.83 years) and 59 healthy, elderly controls (75.37+/-5.27 years). ApoAI levels were significantly lower (p<10(-7)) in AD patients. An apoAI cutoff value of 1.50 g/L, could distinguish between the two groups with a sensitivity of 71% and a specificity of 69%. ApoAI levels were highly correlated with mini-mental state (MMSE) scores of patients (p<0.0001). These relationships remained significant after adjustment for multiple testing. Our findings raise the question of the potential implication of apoAI in the etiopathology of AD and bring serum apoAI concentration to the fore as an important biochemical marker. PMID- 10794846 TI - Alterations in muscarinic receptor-coupled phosphoinositide hydrolysis and AP-1 activation in Alzheimer's disease cybrid cells. AB - Alzheimer's disease cybrid cells produced by replacing endogenous mitochondria in human neuroblastoma SH-SY5Y cells with platelet mitochondria from subjects with Alzheimer's disease have higher levels of reactive oxygen species than do cybrid cells with mitochondria from control subjects. These cells were used to test if this chronic mild increase in reactive oxygen species affects muscarinic receptor coupled signaling activities. Basal and carbachol-stimulated phosphoinositide hydrolysis were higher, and there was less inhibition by glutathione depletion, in Alzheimer's disease than control cybrid cells. Elevated phosphoinositide hydrolysis in Alzheimer's disease cybrid cells also was evident upon direct activation of G-proteins (Gq/11) linked to phosphoinositide signaling or of phospholipase C, but immunoblot analyses revealed equivalent levels of Gq/11 and phospholipase C in both cell lines. These results indicate that there is up regulation of phosphoinositide signaling in Alzheimer's disease cybrid cells in association with chronic mild oxidative stress, although treatment of cells with H(2)O(2) to induce greater acute oxidative stress caused decreases in carbachol stimulated phosphoinositide hydrolysis that were similar in Alzheimer's disease and control cybrid cells. In contrast to phosphoinositide hydrolysis, carbachol stimulated AP-1 DNA binding activity was lower in Alzheimer's disease than control cybrid cells, and this deficit was associated with deficient protein kinase C-mediated activation of AP-1. Overall, these results demonstrate that chronically elevated reactive oxygen species in Alzheimer's disease cybrid cells are associated with a more robust phosphoinositide signaling system, but lower signaling to activation of AP-1. These alterations may represent adaptations to exposure to oxidants, which precede more widespread deficits in signaling associated with more severe oxidative stress. PMID- 10794847 TI - Microglia, amyloid and dementia in alzheimer disease. A correlative study. AB - To elucidate the role of microglia in Alzheimer's disease, a clinicopathological study was performed involving 26 cases, the mental status of which had been studied pre mortem by the Blessed test score (BTS). We measured the volume density of CD 68 immunoreactive (IR) microglia, congophilic plaques and Abeta deposits, and the numerical density of neurofibrillary tangles (NFT) in a sample of Area 9 (middle frontal gyrus). Dementia was significantly correlated only with the volume density of Abeta deposits and the numerical density of NFT. The volume densities of microglia and congophilic plaques were strongly correlated. With the intellectual status used as a time scale, IR microglia and amyloid deposits appeared almost simultaneously at an early stage in the pathological cascade and decreased, whereas Abeta and NFT were still accumulating. The intellectual deficit seemed to be more significantly related to the latter two lesions than to the microglia-amyloid complex, that was visible at an earlier stage (around BTS = 15). PMID- 10794848 TI - Midlife blood pressure and dementia: the Honolulu-Asia aging study. AB - We studied the association of mid-life blood pressure to late age dementia, specifically Alzheimer's disease and vascular dementia. Data are from the cohort of 3703 Japanese-American men who were followed in the Honolulu Heart Program (HHP;1965-1971), and subsequently re-examined in 1991 for dementia. We assessed the risk (odds ratio (95% CI)) for dementia associated with categories of systolic (SBP) and diastolic blood pressure (DBP), stratified by never/ever treatment with anti-hypertensive medications, and adjusting for age, education, apolipoprotein epsilon allele, smoking and alcohol intake. Among those never treated (57% sample), the risk for dementia was OR 95% CI 3.8 (1.6-8.7) for DBP of 90-94 mm Hg, and 4. 3 (1.7-10.8) for DBP of 95 mmHg and over compared to those with DBP of 80 to 89 mm Hg. Compared to those with SBP of 110 to 139 mm Hg, the risk for dementia was 4.8 (2.0-11.0) in those with SBP 160 mm Hg and higher. Blood pressure was not associated with the risk for dementia in treated men. These results were consistent for Alzheimer's disease and vascular dementia. This study suggests elevated levels of blood pressure in middle age can increase the risk for late age dementia in men never treated with anti-hypertensive medication. PMID- 10794849 TI - Midlife blood pressure and neuritic plaques, neurofibrillary tangles, and brain weight at death: the HAAS. Honolulu-Asia aging Study. AB - Midlife hypertension is associated with later development of cognitive impairment, vascular dementia (VsD), and possibly Alzheimer's disease (AD). Neuropathic cerebrovascular lesions and brain atrophy have been associated with elevated blood pressure (BP), however, to our knowledge there have been no prospective investigations of an association of blood pressure levels measured in midlife with the microscopic lesions of AD. We investigated the relationship of BP level in midlife to development of neurofibrillary tangles (NFT), neuritic plaques (NP), and low brain weight at autopsy among Japanese-American men who were members of the Honolulu Heart Program/Honolulu-Asia aging Study (HHP/HAAS) cohort. The HHP/HAAS is a population-based, longitudinal study of cognitive function and dementia with 36 years of follow-up. Neocortical and hippocampal NFT and NP were counted per mm(2), and fixed brain weight was measured for 243 decedents. Elevated systolic BP, (> or =160 mm Hg) in midlife was associated with low brain weight and greater numbers of NP in both neocortex and hippocampus. Diastolic BP elevation, (> or =95 mm Hg) was associated with greater numbers of NFT in hippocampus. Results indicate that in addition to the accepted association of high BP with neuropathic cerebrovascular lesions, there is a direct relationship with brain atrophy, NP and NFT. PMID- 10794850 TI - Brain structure in men remains highly heritable in the seventh and eighth decades of life. AB - The midsagittal cross-sectional dimensions of the corpus callosum, the coronal cross-sectional area of the lateral ventricles at the level of the pons, and a three-dimensional estimate of intracranial volume were derived from magnetic resonance brain images obtained from 45 monozygotic and 40 dizygotic male twin pairs aged 68 to 78. Univariate genetic analyses indicated strong genetic influences contributing significantly to the variability of each brain structure. The estimated proportion of genetic variance (i.e. heritability) was 81% for intracranial volume, 79% for the midline cross-sectional area of the corpus callosum, and 79% for lateral ventricle size. There was no evidence that shared environmental influences contributed significantly to twin-pair similarities. We further used bivariate genetic modeling to estimate the genetic and environmental correlation between correlated brain structures. Intracranial volume and corpus callosum area was highly correlated, and this relationship was entirely due to shared genetic effects between these two brain structures. By contrast, the relationship between the height of the corpus callosum and the size of the lateral ventricles was due to both genetic and environmental influences in common. Corresponding genetic and environmental correlations were 0.68 and 0.58, respectively, indicating that more than half of the genetic and environmental influences on these two brain structures were shared. The manner in which the brain responds to the environment with advancing age is highly genetically determined, both for the lateral ventricles, which dilate with aging and disease, and for the corpus callosum, which is deformed in shape by age-related ventricular enlargement, whereas its midline cross-sectional area remains unchanged. PMID- 10794851 TI - Effect of the angiotensin I-converting enzyme I/D polymorphism on cognitive decline. The EVA Study Group. AB - Several clinical, epidemiological, and pathological observations suggest that vascular risk factors are associated with cognitive performances. The renin angiotensin system components, major determinants of the cardiovascular system, are expressed in the brain. To estimate their potential impact on cognitive performances, we studied the association between cognitive functioning and an insertion/deletion (I/D) polymorphism of the angiotensin I-converting enzyme (ACE) gene. In a sample of 1168 highly performing subjects (59-71 years), DD homozygotes had the lowest cognitive scores as evaluated by the Mini-Mental State Examination. Cognitive decline at 4-year follow-up (defined as the loss of at least 3 points in Mini-Mental State Examination score) was more prevalent in these subjects, the odds ratio being equal to 1.53 (95% CI: 1.04-2.24) with subjects ID as reference class. Moreover, the combined effect of the presence of at least one APOE epsilon4 allele and ACE DD homozygosity was a risk factor for cognitive decline. This report reinforces the hypothesis of an influence of cardiovascular risk factors on cognitive performances. PMID- 10794852 TI - MRI description of cerebral atrophy in mouse lemur primates. AB - We assessed cerebral atrophy in mouse lemur primates (Microcebus murinus) by estimating CSF volume in their brains from 4.7 Tesla T2-weighted magnetic resonance images. Thirty animals aged from 1 to 10.3 years were imaged, 14 of them were followed for up to 2 years. Seven of these animals were examined for neuropathology. In 12 out of 17 animals older than 3.5 years, CSF volumes were increased. A subgroup of six animals had severe atrophy of the temporal lobe. Another subgroup of five animals displayed diffuse atrophy in addition to the temporal atrophy. One animal had a dilation of the external part of the temporal horn of the lateral ventricle in addition to the temporal atrophy. The three animals with diffuse atrophy that could be studied for neuropathology had diffuse cerebral amyloid deposits detected by immunocytochemistry. The other animals did not display amyloid deposits. Relations between the different types of atrophy as well as their causes will have to be assessed in future studies. PMID- 10794853 TI - Region-specific age at onset of beta-amyloid in dogs. AB - Cortical patterns of beta-amyloid (Abeta) deposition were evaluated in 40 beagle dogs ranging in age from 2 to 18 years. Abeta deposition in the prefrontal, occipital, parietal and entorhinal cortices was visualized by using an antibody against Abeta1-42. A logistic regression was used to estimate differences in age at-onset and rate of deposition of Abeta as a function of brain region. The earliest and most consistent site of Abeta deposition with age was in the prefrontal cortex. Entorhinal Abeta deposition was not consistently observed until the age of 14 years, but was present in a subset of dogs under the age of 14 years. These regional vulnerabilities to Abeta accumulation are similar to those seen in the aging human. By using parameters derived from regression analyses, it may be possible to predict the presence of Abeta within specific brain regions in individual dogs. We propose that these models will be a useful tool to evaluate interventions that delay the age of onset or slow the rate of accumulation of Abeta in the dog. PMID- 10794854 TI - Quantitative age-changes in endoplasmic reticulum and nucleus of cerebellar granule cells. AB - A stereological study was performed on cerebellar granule cells from rats 2 to 24 months of age (eight different ages, five animals per age group) to quantify age related alterations in the rough endoplasmic reticulum (RER). The mean surface density and the mean total surface area of the nucleus, as well as the mean absolute volume of euchromatin per cell, were also estimated to examine whether or not these had quantitative relationships with the RER. The mean surface density and the mean total surface area of RER per cell changed significantly, attaining maximum values at 24 months of 1733 microm(2)/1000 microm(3) (0.06) and 64 microm(2) (0.03), respectively, (coefficients of variation in parentheses). The corresponding values at 2 months were 706 microm(2)/1000 microm(3) (0.20) and 26 microm(2) (0.24). The mean absolute volume of the euchromatin changed significantly, with a minimum value of 57 microm(3) (0.05) occurring at 21 months. We postulate that the increase in RER may be part of a mechanism that compensates for an age-related decrease in euchromatin. An increase in the RER network may improve intracellular transport of proteins, production of which is apparently diminished with aging. The increase may also compensate for the reported decrease in calcium buffer capacity of smooth endoplasmic reticulum. PMID- 10794855 TI - Long-term treatment with estrogen and progesterone enhances acquisition of a spatial memory task by ovariectomized aged rats. AB - Female Sprague-Dawley rats were ovariectomized at 13 months of age. Four groups received different regimens of estrogen or estrogen plus progesterone replacement beginning either immediately, 3 months, or 10 months after ovariectomy and were compared with non-hormone-treated controls. Eight to twelve months after ovariectomy, animals were trained on a delayed matching-to-position (DMP) spatial memory task. Long-term treatment with estrogen or estrogen plus progesterone significantly enhanced acquisition of the DMP task by aged animals after long term loss of ovarian function. Weekly administration of estrogen and progesterone was at least as effective as, if not more effective than, continuous treatment with estrogen alone. In addition, treatment initiated 3 months, but not 10 months, after ovariectomy was as effective at enhancing DMP acquisition as continuous estrogen treatment initiated immediately after ovariectomy, suggesting a window of opportunity after the loss of ovarian function during which hormone replacement can effectively prevent the effects of aging and hormone deprivation on cognitive function. These findings suggest that repeated treatment with estrogen and progesterone initiated within a specific period of time after the loss of ovarian function may be effective at preventing specific negative effects of hormone deprivation on brain aging and cognitive decline. PMID- 10794856 TI - Effects of intra-striatal GDNF on motor coordination and striatal electrophysiology in aged F344 rats. AB - The purpose of this study was to examine whether improvement in motor function could be demonstrated in old rats, and to see if GDNF affected post-synaptic DA function. Aged (20 month old) versus young rats were tested following GDNF treatment for postural control by using an inclined balance beam and a wire grip strength test. Rats were also examined electrophysiologically for spontaneous striatal cell firing rate alone and in the presence of DA receptor agonists, and histologically for the intensity of striatal TH staining, and number of DA containing nigral cells. Behavior was significantly improved in the aged animals who received central GDNF infusions, although the extent of improvement was less than what has been observed in 16-month-old rats. There was no effect of GDNF treatment in the aged animals on spontaneous firing rate in the striatum, or on the post synaptic response to locally applied D(1) and D(2) receptor family agonists. However, there was an effect of age alone on firing rate, and on the response to locally applied SKF 38393 and quinpirole. By using unbiased cell counting we observed no age-related decline in the number of TH positive cells in the substantia nigra. There was no effect of GDNF on the number of TH positive cells in the substantia nigra in either young or aged rats, although there were morphological improvements in DA neurons of the GDNF treated aged rats. These results replicate earlier studies showing an effect of age on striatal firing rate and dopamine receptor function, and suggest that the GDNF mediated improvement in behavior may be located other than post synaptically within the striatum. PMID- 10794857 TI - Cholinergic medial septum neurons do not degenerate in aged 129/Sv control or p75(NGFR)-/-mice. AB - Cholinergic medial septum neurons express TrkA and p75 nerve growth factor receptor (p75(NGFR)) and interactions between TrkA and p75(NGFR) are necessary for high-affinity binding and signaling of nerve growth factor (NGF) through TrkA. In adult p75(NGFR)-deficient (-/-) mice, retrograde transport of NGF and other neurotrophins by these neurons is greatly reduced, however, these neurons maintain their cholinergic phenotype and size. Reduced transport of NGF has been proposed to play a role in Alzheimer's disease. Here, we investigated whether chronic and long-term absence of p75(NGFR) (and possibly reduced NGF transport and TrkA binding) would affect the cholinergic septohippocampal system during aging in mice. In young (6-8 months), middle aged (12-18 months), and aged (19-23 months) 129/Sv control mice the total number of choline acetyltransferase positive medial septum neurons and the mean diameter and cross sectional area of the cholinergic cell bodies were similar. The cholinergic hippocampal innervation, as measured by the density of acetylcholinesterase-positive fibers in the outer molecular layer of the dentate gyrus was also similar across all ages. These parameters also did not change during aging in p75(NGFR) -/- mice and the number and size of the choline acetyltransferase-positive neurons and the cholinergic innervation density were largely similar as in control mice at all ages. These results suggest that p75(NGFR) does not play a major role in the maintenance of the number or morphology of the cholinergic basal forebrain neurons during aging of these mice. Alternatively, p75(NGFR) -/- mice may have developed compensatory mechanisms in response to the absence of p75(NGFR). PMID- 10794858 TI - Effect of tacrine on EEG slowing in the rat: enhancement by concurrent monoamine therapy. AB - A dominant electrophysiological characteristic of Alzheimer's disease (AD) is the loss of desynchronized EEG activity and shift toward low-frequency EEG synchronization. In rats, similar EEG changes resulted from administering the anti-cholinergic scopolamine (1 mg/kg) and the monoamine depletor reserpine (10 mg/kg); amplitude increases between 0.5-20 Hz, with the delta (0.5-4 Hz) and theta (4-8 Hz) bands affected most severely. The acetylcholinesterase inhibitor tacrine, at doses between 10 and 20 mg/kg, reversed these EEG changes; co administration of tacrine and the noradrenaline-serotonin reuptake inhibitor imipramine (10 mg/kg) enhanced tacrine's action to suppress delta activity. Co administration of tacrine and the monoamine-oxidase inhibitor pargyline (20 mg/kg) enhanced EEG restoration by tacrine in all frequency bands between 0.5 to 20 Hz, but co-administration of the selective serotonin reuptake inhibitor fluoxetine (2 mg/kg) was ineffective. These results show that some drug therapies aimed at concurrently stimulating cholinergic and monoaminergic neurotransmission are more effective in reversing EEG slowing than cholinergic therapy alone. Significant monoaminergic deficits occur in Alzheimer's disease, in addition to the atrophy of cholinergic neurons. Thus, combined cholinergic-monoaminergic therapy may provide an enhanced restoration of cortical functioning, in addition to limiting the required treatment dose of cholinesterase inhibitors. PMID- 10794859 TI - Dopamine and senescence in Drosophila melanogaster. AB - Five-day-old Drosophila melanogaster males, when exposed to 2-h-old males, will perform courtship rituals; the intensity and duration of this behavior rapidly diminishes with time. The ability of the older males to habituate to the attractive signals given off by the younger males is a dopaminergic-modulated experience-dependent modification of behavior that is abolished with increasing age. Dopamine-depleted females show increased resistance to copulation; 20-day old females demonstrated an increase in copulation avoidance compared with younger (5-15-day-old) females. These changes in dopaminergic-modulated behaviors observed during aging parallel declines in whole body levels of dopamine. Immunocytochemical analysis of adult brains using an antibody raised against Drosophila tyrosine hydroxylase to visualize catecholaminergic cell bodies revealed increased degeneration of the cell bodies with aging. These results suggest that the deficits seen in dopaminergic-modulated behaviors may arise as a consequence of degenerative changes within the aging brain. PMID- 10794861 TI - Transducer materials and probe construction. PMID- 10794862 TI - Intravascular scanners. PMID- 10794863 TI - 2-D arrays. PMID- 10794865 TI - Color flow mapping. PMID- 10794864 TI - Doppler signal analysis. PMID- 10794866 TI - Nonlinear imaging. PMID- 10794867 TI - Elastographic imaging. PMID- 10794868 TI - Microscopy. PMID- 10794869 TI - Contrast agents. PMID- 10794870 TI - Three-dimensional imaging. PMID- 10794871 TI - Image segmentation in obstetrics and gynecology. PMID- 10794872 TI - Spectroscopy and image texture analysis. PMID- 10794873 TI - Telesurgery. PMID- 10794874 TI - Quality assurance for grey-scale imaging. PMID- 10794875 TI - Intervention and therapy. PMID- 10794876 TI - Lithotripsy. PMID- 10794877 TI - Gene transfection and drug delivery. PMID- 10794878 TI - Standards and measurement. PMID- 10794879 TI - Biophysical aspects of diagnostic ultrasound. PMID- 10794880 TI - Hepatobiliary system. PMID- 10794881 TI - Splanchnic vessels. PMID- 10794882 TI - Urinary system. PMID- 10794883 TI - Endoluminal gastroenterology. PMID- 10794884 TI - Obstetrics and perinatology. PMID- 10794885 TI - Paediatrics. PMID- 10794886 TI - Gynecology. PMID- 10794887 TI - Peripheral vascular system. PMID- 10794888 TI - Extracranial vessels. PMID- 10794889 TI - Pediatric neurology. PMID- 10794890 TI - Adult cardiology. PMID- 10794891 TI - Pediatric cardiology. PMID- 10794892 TI - Breast. PMID- 10794893 TI - Superficial structures. PMID- 10794894 TI - Male genital system. PMID- 10794895 TI - Musculoskeletal system. PMID- 10794896 TI - Ophthalmology. PMID- 10794897 TI - Interventional procedures. PMID- 10794898 TI - Fetal intervention. PMID- 10794899 TI - 2-D and 3-D endoluminal ultrasound. PMID- 10794900 TI - Intraoperative procedures. PMID- 10794902 TI - Industrial perspective. PMID- 10794901 TI - Microgravity. PMID- 10794903 TI - Africa. PMID- 10794904 TI - Asia. PMID- 10794905 TI - Australia and New Zealand. PMID- 10794906 TI - Developing countries. PMID- 10794907 TI - Europe. PMID- 10794908 TI - Latin America. PMID- 10794909 TI - North America. PMID- 10794910 TI - The variation of mass and disinfection by-product formation potential of dissolved organic matter fractions along a conventional surface water treatment plant. AB - Dissolved organic matter (DOM) influences many aspects of water treatment, including the formation of potentially harmful disinfection by-products (DBPs) when disinfectants are applied. DOM from a conventional surface water treatment plant (WTP) in Northern New Jersey was isolated and fractionated using resin adsorption chromatography into six different fractions. These fractions are operationally categorized as hydrophobic acid, hydrophobic neutral, hydrophobic base, hydrophilic acid, hydrophilic neutral and hydrophilic base. The hydrophilic acid fraction was found to be the most abundant fraction in the source water. The hydrophilic neutral, hydrophilic acid and hydrophobic acid fractions had the highest removal efficiency through the WTP (about 65%). The variation and removal effectiveness of each fraction along the WTP was studied. Seven-day chlorine DBP formation potential (FP) tests were performed on all DOM fractions through the WTP. For the source water studied, the hydrophilic acid fraction was found to be the most reactive precursor to the trihalomethane (THM) formation. The hydrophobic neutral fraction was found to be the fraction of concern with respect to the FP of haloacetic acids (HAAs) class of DBPs. The FP of each fraction's class of DBPs was found to be amenable for reduction along the treatment train, specifically by coagulation/sedimentation. The fractionated approach concept showed to be very beneficial in the study of DBP precursors and their effective removal by physical and chemical treatment. PMID- 10794911 TI - Fire and explosion hazards to flora and fauna from explosives. AB - Deliberate or accidental initiation of explosives can produce a range of potentially damaging fire and explosion effects. Quantification of the consequences of such effects upon the surroundings, particularly on people and structures, has always been of paramount importance. Information on the effects on flora and fauna, however, is limited, with probably the weakest area lying with fragmentation of buildings and their effects on different small mammals. Information has been used here to gain an appreciation of the likely magnitude of the potential fire and explosion effects on flora and fauna. This is based on a number of broad assumptions and a variety of data sources including World War II bomb damage, experiments performed with animals 30-40 years ago, and more recent field trials on building break-up under explosive loading. PMID- 10794912 TI - Field monitoring and performance evaluation of an in situ air sparging system at a gasoline-contaminated site. AB - In situ air sparging (IAS) has been used since the mid-1980s, but few carefully designed field studies have been performed to evaluate its effectiveness. In this study, 27 discrete monitoring points (MPs) were installed at a gasoline contaminated site to investigate the efficacy of IAS. Each MP was instrumented with a pressure transducer and a Technalithics dissolved oxygen (DO) probe, and located so they could be used to characterize subsurface changes in total head and DO with depth, distance and orientation around a central injection well. Because the blower over-heated and automatically shut down after approximately 30 min and short-circuiting of air into two MPs occurred within 2 min, the study was designed as three sets of three 30-min trials. Longer trials would not have yielded different nor more insightful results. A volume of soil was not oxygenated during any injection. Instead, air traveled directly to at least four of seven different MPs during eight of the nine trials, probably as a result of an air bubble forming beneath a confining layer. The order of air arrival at the MPs varied during the first few trials, but once a preferential pathway was established, it did not collapse between trials and provided the shortest distance to the vadose zone during subsequent trials. Oxygen uptake rates estimated for MPs that received air during any trial exceeded the consumption rates of the Technalithics DO probes, and indicate that the probes could be used for estimating oxygen transfer during system operation or for oxygen uptake measurements during shut-down tests. The data from the monitoring system indicate that IAS is infeasible for remediation of soil and groundwater at this site due to its low horizontal hydraulic conductivity. Similar behavior is anticipated when IAS is applied at other sites with low hydraulic conductivity materials. PMID- 10794913 TI - Decomposition of isoquinoline and quinoline by supercritical water. AB - The ability of supercritical water (SCW) to decompose heterocyclic compounds (quinoline and isoquinoline) has been explored in this study. The results obtained suggest that water acts as a chemical reagent above its critical point (374 degrees C and 22.1 MPa). Significant proportions of isoquinoline and quinoline were removed during the reaction with SCW. The response of these compounds to pyrolysis was also compared with their reaction with SCW. Both compounds were relatively more reactive in the presence of SCW than during pyrolysis. Because of the different positions of N atom in the two compounds, they reacted with SCW differently. Breaking of C-N bonds during SCW reaction was by hydrogenation and hydrocracking, while pyrolysis was due to thermocracking mainly. PMID- 10794914 TI - Enhanced TCDD degradation by Fenton's reagent preoxidation. AB - The dioxin isomer 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been called the most toxic compound known to man. Because of its poor bioavailability and low biodegradibility, bioremediation technology cannot effectively degrade TCDD when used alone. In this study, chemical pretreatment (partial oxidation) in combination with biodegradation technique was developed to efficiently remediate TCDD-contaminated soils. An oxidizing reagent [Fenton's Reagent (FR)] was applied in a slurry reactor to transform TCDD with a concentration of 96 microg per kg of soil to compounds more amenable to biodegradation. Up to 99% TCDD was transformed after the chemical pretreatment process. The slurry reactor was then converted to a bioreactor for the following biodegradation experiment. The detected TCDD oxidation byproducts including chlorophenols (CPs) and chlorobenzenes (CBs) were transformed in this bioreactor under aerobic conditions. Two other biodegradation experiments were performed in parallel to investigate the biodegradabiliy of TCDD under aerobic and anaerobic conditions without chemical pretreatment. Approximately 53% TCDD was transformed under anaerobic conditions possibly due to the reductive dechlorination process using organic materials contained in the activated sludge as the primary substrates. No TCDD degradation was observed under aerobic conditions. Results show that FR can oxidize TCDD to less chlorinated and less-toxic byproducts, promoting their bioavailability to microbial communities. The bench-scale results indicate that the two-stage (partial oxidation followed by biodegradation) system has the potential to be developed to remediate TCDD-contaminated soils on-site. PMID- 10794915 TI - Behavioral rise in body temperature and tachycardia by handling of a turtle (Clemmys insculpta). AB - Three turtles, Clemmys insculpta, were kept together in a terrarium in a climatic chamber at 18 degrees C, with lights on at 07:00 h and off at 19:00 h. In one corner of the terrarium an infrared lamp produced an operative temperature of 42.5 degrees C, thereby allowing behavioral temperature regulation during the light period. When the turtles were handled only once a day for the purpose of taking cloacal temperature, their body temperature held stable at about 22-23 degrees C. Immediately after being handled the turtles sought the radiant heat and regulated their body temperature at about 4 degrees C higher than before the handling. When repeatedly handled every 15 min for 2 h the turtles maintained a high body temperature by their behavior. When not repeatedly handled the turtles returned to their initial preferred body temperature ca 22-23 degrees C within 2 h. It is hypothesized that handling causes in turtles a fever similar to that observed in stressed mammals. The turtles were equipped with an electrocardiogram radio transmitter and their heart rate was recorded at a distance. Heart rate in undisturbed turtles was 28.3+/-0.6 bt/min. During a 1-min handling, their heart rate rose to 40.2+/-0.8 bt/min. This tachycardia persisted several minutes, then their heart rate returned to the baseline value in ca. 10 min. Stress fever and tachycardia are taken as signs of emotion in turtles. PMID- 10794916 TI - Use of cues by Lipophrys pholis L. (Teleostei, Blenniidae) in learning the position of a refuge. AB - The ability of Lipophrys pholis to remember the position of a refuge was tested in an artificial habitat under the influence of different visual clues. L. pholis learned the position of the refuge in the presence of a clue consisting only of a small black screen. They responded to this clue by moving towards it and pressing themselves up against it. Lego towers and a white screen clue did not provoke such a response. In a further experiment L. pholis continued to respond to the black screen in this way when the screen was moved to another location further from the refuge. After 12 days L. pholis learned to use the black screen in its new position as an indirect clue and navigate to the refuge directly without first approaching the black screen. These results suggested that when placed in a novel habitat the immediate reaction of L. pholis is to move quickly towards the first dark area they see but, with experience, they can use the position of large objects around them to navigate quickly and efficiently to a refuge. PMID- 10794917 TI - Effect of environmental enrichment upon resource holding power in fish in prior residence situations. AB - Resource holding power (RHP), as expressed by gaining dominance, can be affected by extrinsic and intrinsic factors. Extrinsic factors that increase the RHP include e.g. prior exposure to the contest area. The pay-off asymmetry hypothesis was tested according to which there is an asymmetry in the value the resident's territory has for the resident and intruders, i.e. the resident loses more than intruder opponents when losing the contest over a territory in which it has invested much energy during the exploration of food and shelter resources, during the settlement of conflicts with territorial neighbours, etc. This asymmetry will increase the resident's chance to win the fight over intruders. If the resident intruder asymmetry is enhanced by presenting resource-rich territories to e.g. fish (structurally diverse or 'rich' aquariums), the hypothesis predicts that the resident's dominance chance (probability to win the fight) will be higher than in structurally 'poor' aquariums. It was found that residents were proportionately more often dominant (fight winners) than intruders in the rich compared to the poor aquariums in all seven tested fish species. This demonstrates that a high territory resource value (aquarium enrichment) significantly facilitates the expression of the resident's dominance advantage (prior residence effect). In contrast to dominance, aggressive behaviour before the dominance settlement did not generally differ between residents and intruders when compared in rich and poor aquariums. This suggests that dominance in itself and aggression prior to dominance settlement are at least partially guided by different motivational systems. PMID- 10794918 TI - The effect of delayed feeding on the post-feeding behaviour of sows. AB - Previous research into oral persistence in pigs has shown that the relationship between motivation and performance of stereotypy is not a simple one, and that non-specific motivational factors such as arousal may be important. For chronically food-restricted sows the time before food arrives is characterised by increasing excitement and arousal which may be carried over into the post-feeding period facilitating the performance of persistent oral behaviour. To investigate this possibility, arousal was manipulated by delaying feeding. Five and 15 min delays were used and sows experienced delayed feeding 1 day in every 4. Principal components analysis (PCA) was used to group the behaviour data into two factors, ACTIVITY/CHEW and NOSE/ROOT/PEN and these scores were then analysed using ANOVA as well as the appropriate individual behaviours. The delay of feeding increased the rate of eating, presumably reflecting increased arousal (P<0.05). ACTIVITY/CHEW scores were highest in 5 min delay sets, and an interaction between day of set and delay length indicated that ACTIVITY/CHEW scores peaked on different days within a set (P<0.05). Time spent chewing was highest on the day of delay (P<0.05), while standing was highest in days 1 and 2 post-delay (P<0.05). The NOSE/ROOT/PEN scores were lower on the day of delay than on other days (P<0.05), and were lowest in the first delay set than in sets later in the trial (P<0.01). It appeared that increasing arousal in the pre-feeding period does affect activity and oral behaviours in the post-feeding period. However, some of the effects appeared in a more extended and diffuse manner, perhaps due to the general disturbance created by the experimental regime and the sows' experience of long-term food restriction. PMID- 10794919 TI - Acoustic pattern variations in the female-directed birdsongs of a colony of laboratory-bred zebra finches. AB - The acoustic profile of the zebra finch song is characterized by a series of identical repeating units, each comprising a distinctive sequence of acoustic elements, called syllables. Here, we perform an analysis of song pattern deviations caused by variabilities in the production of song syllables. Zebra finches produce four different kinds of syllable variabilities-syllable deletions, single or double syllable insertions, syllable alterations, and syllable repetitions. All these variabilities, with the exception of repetitions, are present in songs of more than two-thirds of the normal adult birds; repetitions are present in less than one-fifth of birds. The frequency of occurrence of these variabilities is independent of the amount of singing, suggesting that they are unlikely to result simply from singing-induced physiological changes such as fatigue. Their frequencies in tutor-deprived birds are not significantly different from those in normal birds, indicating that they are unlikely to be acquired due to deficiencies in tutor-dependent learning. The types, patterns of occurrence and relative frequencies of these song syllable variabilities might reveal insights into the functioning of the song motor control pathway. PMID- 10794920 TI - Preferences of farmed blue foxes for different floor types. AB - In the first experiment, farmed blue foxes were allowed to choose for 1 week between four standard farm cages equipped with different floor materials: plastic coated wire mesh, dry wood, dry sand and wet (summer) or icy (winter) sand. Resting consisted of 10-15 separate bouts/day occupying 50-60% of the total 24-h. There were no other differences in the use of the cages except that the time spent on, and the duration and number of resting bouts were lower on wet or icy sand, resting periods being more affected than activity. In the second experiment, two cages were connected with a 1.2 m tunnel. One cage was always elevated (50 cm) compared to that one which was lower. One cage of each pair had a wire floor whereas the other cage either had sand or wire floor. Sand floor was preferred for active behaviours and wire floor for resting if these were on elevated level. Of two identical wire-floored cages, the elevated one had the priority. Foxes preferred to rest on the same floor where they had finished their previous resting bout, often exclusively and independently of floor material or floor level. PMID- 10794921 TI - Effect of penetration enhancers on isosorbide dinitrate penetration through rat skin from a transdermal therapeutic system. AB - Percutaneous absorption of isosorbide dinitrate (ISDN) from a transdermal therapeutic system (TTS) with or without penetration enhancers was studied. The concentration of ISDN and its metabolites, isosorbide-5-mononitrate (IS-5-MN) and isosorbide-2-mononitrate (IS-2-MN), was determined in rat plasma during a 48 h application of TTS. The increased skin-penetration enhancing effect of oleic acid and propylene glycol in comparison to polyethylene glycol 400 and isopropyl myristate on percutaneous permeation of ISDN was shown. It was expressed by higher values of C(max) and AUC. After the application of TTS, a lower ISDN and molar ratio of its metabolites was observed than after oral administration. PMID- 10794922 TI - Mapping PET-measured triamcinolone acetonide (TAA) aerosol distribution into deposition by airway generation. AB - The three dimensional (3D) distribution of inhaled drugs was measured using Positron Emission Tomography (PET) (Berridge, M.S, Muswick, G.J., Lee, Z., Leisure, G.L., Nelson, A.D., Muzic, R.F. Jr., Miraldi, F., Heald, D.L., 1997. PET evaluation of Azmacort(R) ([C-11]triamcinolone acetonide) dose administration. J. Nucl. Med. 38 (5) Suppl., 4-5). Data analysis was based upon regional ratios or penetration indices. To improve the analytical usefulness and objectivity, labeled drug from dynamic PET images was mapped into 23 airway generations following a general framework from a SPECT-based methodology (Fleming, J.S., Nassim, M.A., Hashish, A.H., Bailey, A.G. , Conway, J., Holgate, S., Halson, P., Moore, E., Martonen, T.B., 1995. Description of pulmonary deposition of radiolabeled aerosol by airway generation using a conceptual three dimensional model of lung morphology. J. Aerosol Med. 8, 341-356). A recently developed airway network model was used in this study. Quantitative PET scans of [C 11]triamcinolone acetonide distribution in the lung were determined following administration of Azmacort(R), a commercial metered dose inhaler with an integrated spacer device. Distributions at varying time periods after drug administration were investigated to explore the dynamics and kinetics of the aerosolized drug. Initially, deposition of labeled drug on conducting airways (generations 1-14) was found to be higher than those on acinar airways (generation 15-23), 64% versus 36%. The distribution pattern changed slowly with time. By 47 min, 51% of the dose remaining in the lung was found on conducting airways while 49% was on acinar airways. This study illustrates the value of PET imaging for the evaluation and design of drug formulations. PMID- 10794923 TI - Factors limiting the oral bioavailability of N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) and enhancement of absorption in rats by delivery in a water-in oil microemulsion. AB - The bioavailability (BA) of radio-labelled N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) was low when administered by oral gavage as an aqueous solution to conscious male Sprague-Dawley rats (8.3+/-4.4% (mean+/-S.D., n=3)). To assess the likely factors contributing to the poor BA of GMDP, the stability of GMDP in the lumen of the gastrointestinal (GI) tract was examined in vitro, using ex vivo GI contents. GMDP was degraded by the contents of the small intestine, caecum and large intestine but was more stable in stomach contents. The permeability coefficient (p(app)) of GMDP in isolated sections of rabbit ileum was 1.67x10(-6) cm/s in the mucosal to serosal direction and was not significantly different in the serosal to mucosal direction, indicating that GMDP is poorly permeable and passively transported across the intestinal wall. First pass metabolism was considered to be unlikely to be the primary limitation to the oral bioavailability of GMDP and therefore, that the oral bioavailability of GMDP was likely limited by instability in the lumen of the gastrointestinal tract and low intestinal permeability. A water-in-oil (w/o) microemulsion formulation subsequently developed to address these problems was trialed in a preliminary bioavailability study in rats and enhanced the bioavailability of GMDP ten-fold when administered intraduodenally, indicating that w/o microemulsions may represent a viable mechanism for enhancing the bioavailability of poorly GI stable and poorly permeable peptide-based molecules. PMID- 10794924 TI - Physical evaluation of a new patch made of a progestomimetic in a silicone matrix. AB - The adhesion of a new Transdermal Therapeutic System (TTS) made of silicone and loaded with a progestomimetic drug was characterised. The goal of this study was to use well-known methods or to adapt them to collect representative data. Individually, methods such as surface tension, peel test and rheology are already widely used. Results show that the choice of a substrate for peel tests can be made in the light of surface tension data and that polymers like poly(tetrafluoroethylene) (PTFE) are good alternatives to skin. Peeling characterisations are made a function of thickness of films, drug content in active, conditions of preparation and conditions of use such as pressure. Dynamic rheology is more difficult to link to other methods as it mainly reflects internal phenomena and properties that arise in the bulk, as opposed on its surface. Master curves enable results to be used more easily, but the theories to interpret the data are still not powerful enough to replace peel testing. PMID- 10794925 TI - In vitro and in vivo evaluation of caffeic and ferulic acids as topical photoprotective agents. AB - Topically-applied antioxidant drugs represent a successful strategy for protecting the skin against UV-mediated oxidative damage. However, they can afford to the skin a satisfactory photoprotection only if able to permeate through the stratum corneum and thus to reach deeper cutaneous layers. Caffeic and ferulic acids, dissolved in saturated aqueous solutions at pH 3 or 7.2, have been tested for their capability to permeate through excised human skin mounted in Franz cells. At both pH values, ferulic and, at a lower degree, caffeic acids appeared able to permeate through the stratum corneum. The known higher lipophilicity of ferulic acid may explain the fact that it permeates through the stratum corneum better than caffeic acid. However, vehicle pH values proved to have no influence on biophenol skin permeation profile; this observed lack of pH effect may reflect the drug higher concentration attainable in saturated solutions at high pH. On the basis of the findings obtained in these in vitro experiments, we designed the schedule of a series of in vivo experiments, carried out to evaluate the ability of caffeic and ferulic acids to reduce, in healthy human volunteers, UVB-induced skin erythema, monitored by means of reflectance spectrophotometry. Caffeic and ferulic acids, dissolved in saturated aqueous solution pH 7.2, proved to afford a significant protection to the skin against UVB-induced erythema. To conclude, we have confirmed, by means of in vitro and in vivo experiments, that caffeic and ferulic acids may be successfully employed as topical protective agents against UV radiation-induced skin damage; however their skin absorption is not influenced by the pH of the formulation. PMID- 10794926 TI - Investigation on the photostability of a tretinoin lotion and stabilization with additives. AB - Tretinoin, a drug that is used in topical preparations for the treatment of acne vulgaris, is known to be very susceptible to degradation under daylight. The objective of this work was to investigate the degradation of a tretinoin lotion placed in front of a xenon lamp. Analysis was performed with HPLC. The tretinoin lotion was degraded to about 20% of its initial concentration within 30 min. Incorporation of tretinoin in beta-cyclodextrin or in some surfactants (Brij(R)s) did not have any effect on the photodegradation of tretinoin. Neither could a UV B sunscreen retard the photodegradation of tretinoin while a UV-A sunscreen had very little effect. Irradiation with selected wavelengths revealed that 420 nm seemed to be the most harmful wavelength for the degradation of tretinoin and not the wavelength of maximum absorption (350 nm) as expected. Then the addition of the yellow colourants chrysoin and fast yellow, absorbing in the region of 420 nm, was tested. These colourants did indeed retard the photo-degradation of tretinoin more or less depending on the concentration of the dye. Finally we only had to select a concentration that was still effective but that did not colour the skin. PMID- 10794927 TI - Conformational analysis of 7-oxoacyl-L-alanyl-D-isoglutamines. AB - In a recent article (Planinsek, O., Srcic, S., 1999. Int. J. Pharm. 87, 199-207) some interesting physicochemical properties of a series N-(7-oxoacyl)-L-alanyl-D isoglutamines with n=0-6 methylene groups between the terminal methyl and 7-oxo group were measured. In view of the practical importance of these N acetylmuramyldipeptide(MDP) immunomodulator analogues and their interesting biological properties a detailed conformational analysis was undertaken for the series with n=3-6 methylene spacers between the 7-oxo and terminal methyl groups. The puzzle posed by the reversal of the measured water solubility and lipophylicity could be resolved by using the Monte Carlo approach to searching the conformational space of the molecules in this series. We have found that the increase in water solubility and drop in lipophylicity when the number of methylene groups is increased from 5 to 6 can be attributed to the change in predominant conformation in the conformational family as described by the Boltzmann distribution of conformations. Notwithstanding this, we point out the changes in biological response coupled to the nonlinearity of the physicochemical behaviour in the series. PMID- 10794928 TI - Rapid-onset intranasal delivery of anticonvulsants: pharmacokinetic and pharmacodynamic evaluation in rabbits. AB - Intranasal (IN) administration is a promising approach for rapid-onset delivery of medications. In order to assess the feasibility of this approach for the emergency treatment of status epilepticus, three anticovulsants, i.e. diazepam (DZ), clonazepam (CZ), and a monocarbamate-based new compound (MCA) were studied in rabbits for the pharmacokinetics (PK) and pharmacodynamic (PD) response following intravenous (IV) and IN administrations. The animals were intranasally dosed with DZ (1 mg/kg), CZ (0.2 mg/kg), and MCA (5 mg/kg), dissolved in 200 microl of vehicle consisting of propylene glycol (PG), ethanol (EtOH), and water in the presence or absence of 1% sodium glycocholate (SGC) using single and repeated dosing schedules. Both DZ and CZ were absorbed very rapidly from 1% SGC/60% PG-30% EtOH-10% Water after IN single application; the T(max)'s were less than 2 min. The absorption rate of MCA was relatively slower with the peak time of 13-32 min. The bioavailability of single IN administration for DZ, CZ, and MCA determined over the first 2 or 4 h was found to be 77, 45, and 79%, respectively. The peak plasma level of DZ increased linearly with increasing the volume fraction of EtOH in the ternary cosolvent (20% to 60%). A repeated IN application of DZ, 5 min after the first dose, doubled the C(max) and AUC(0-2 h) values of the first one, whereas those of CZ and MCA resulted in an increase of 73-94% of the first dose. A single IN application of DZ- and CZ-containing formulations produced a PD response within 1.5 min, which was comparable to that of an IV injection. These results suggest that single or repeated IN applications of DZ, CZ, and MCA in a hydroalcohol-glycolic formulation might represent a viable approach to achieving a rapid systemic absorption of these anticonvulsants during the emergency treatment of status epilepticus and other types of seizures. PMID- 10794929 TI - Application of Carbopol to controlled release preparations I. Carbopol as a novel coating material. AB - We investigated the application of Carbopol(R) (CP) as a novel coating material prepared with various grades of CP having different degrees of cross-linking and molecular weights. Viscosity and spray mist size of CP aqueous solutions at various concentrations of CP were measured. Core tablets containing theophylline (TP), as a model drug, were coated with CP at various coating ratios. The TP release profile from the CP-coated tablets was studied by the JP13 paddle method. CP tablets were prepared by compressing CP powder, and the swelling behavior of the CP tablets in JP 1st fluid, purified water, and JP 2nd fluid was observed. The spray mist size of all CP aqueous solutions was small at a concentration of 1% and below, and drastically increased over a concentration of 1%. This result suggests that the appropriate concentration of the CP solution for coating is 1% or below. Sustained release of TP from the CP-coated tablets at a coating ratio of only 3% was observed in the JP 1st fluid and purified water, although fast release was observed in the JP 2nd fluid. The fast release in the latter fluid may be due to the fact that CP is an acid material. These results suggest that it is feasible to control the drug release by use of an extremely small amount of CP coating and that CP is useful as a novel coating material. PMID- 10794930 TI - Controlled release injectable liposomal gel of ibuprofen for epidural analgesia. AB - The epidural administration is used commonly in the treatment of pain. Nonsteroidal anti-inflammatory drugs, especially ibuprofen, would have potential in epidural use. Like many epidurally useful drugs it, however, has a short duration of action, which is a limiting factor. To improve epidural pain treatment, a long-acting, single-dose gel injection is being developed. In the present study, the possibility of using liposomal systems to control the release and dural permeation of ibuprofen was investigated in vitro. Liposomal solutions of ibuprofen.Na (20 mg/ml) were prepared by high-pressure homogenization from egg phosphatidylcholine. The liposomal gel consisted of poloxamer 407 and the liposomal solution. No signs in the 1H-NMR spectroscopy of line broadenings or chemical shifts were observed. The liposomal formulations were reproducible and stable. Ibuprofen release in phosphate buffer, pH 7.4, at 37 degrees C from the liposomal solution and the liposomal gel were prolonged significantly compared with their respective solution and gel controls. The liposomal gel controlled ibuprofen release and dural permeation in vitro and showed a permeation pattern favourable for maintaining constant drug levels. The liposomal poloxamer gel represents a new formulation approach to increase the local epidural availability of ibuprofen. It appeared to be a promising injectable controlled-release drug delivery system. PMID- 10794931 TI - Defining the drug incorporation properties of PLA-PEG nanoparticles. AB - The drug incorporation and physicochemical properties of PLA-PEG micellar like nanoparticles were examined in this study using a model water soluble drug, procaine hydrochloride. Procaine hydrochloride was incorporated into nanoparticles made from a series of PLA-PEG copolymers with a fixed PEG block (5 kDa) and a varying PLA segment (3-110 kDa). The diameter of the PLA-nanoparticles increased from 27.7 to 174.6 nm, with an increase in the PLA molecular weight. However, drug incorporation efficiency remained similar throughout the series. Incorporation of drug into the smaller PLA-PEG nanoparticles made from 3:5, 15:5 and 30:5 copolymers did not influence the particle size, while an increase was observed for the larger systems comprising 75:5 and 110:5 copolymers. An increase in drug content for PLA-PEG 30:5 nanoparticles was achieved by increasing the theoretical loading (quantity of initially present drug). The size of these nanoparticles remained unchanged with the increasing drug content, supporting the proposed micellar type structure of the PLA-PEG 30:5 nanoparticles. The morphology of these systems remained unchanged both at low and high theoretical drug loadings. Formulation variables, such as an increase in the aqueous phase pH, replacement with the base form of the drug and inclusion of lauric acid in the formulation did not improve the incorporation efficiency of drug into PLA-PEG 30:5 nanoparticles. While poly(aspartic acid) as a complexation agent did not improve the drug incorporation efficiency of procaine hydrochloride, it did so for another water soluble drug diminazene aceturate. This may be attributed to a stronger interaction of diminazene aceturate with poly(aspartic acid) relative to procaine hydrochloride, as confirmed by thermodynamic analysis of isothermal titration calorimetric data. The drug incorporation and physicochemical characterisation data obtained in this study may be relevant in optimising the drug incorporation and delivery properties of these potential drug targeting carriers. PMID- 10794932 TI - Performance of a multiplex PCR for the determination of Haemophilus influenzae capsular types in the clinical microbiology laboratory. AB - To improve tools for the surveillance of invasive H. influenzae in the context of the drastic decrease of type b infections following the implementation of vaccination, a two-step PCR technique was developed to detect the capsule and type specific regions of H. influenzae. The technique of Falla et al (1994) was modified to amplify in a first step the capsule and type b regions by multiplex PCR. For non-b capsulated strains, the type a, c, d, e, and f loci were afterward detected simultaneously by an optimized touch-down PCR technique. An internal control of extraction and amplification (16S rDNA) was included for both PCR techniques. Overall, this technique was shown to perform as efficiently or better than the slide agglutination without risks of interpretation errors. Of the 138 H. influenzae strains tested, seven that had given doubtful results by the agglutination technique were unequivocally typed by PCR. PMID- 10794933 TI - Evaluation of the BacT/Alert microbial detection system with FAN aerobic and FAN anaerobic bottles for culturing normally sterile body fluids other than blood. AB - We evaluated the BacT/Alert Microbial Detection System (Organon Teknika Corporation, Durham, NC, USA) by using FAN bottles compared to conventional culture methods for the recovery of microorganisms from normally sterile body fluids other than blood and dialysates. Clinically significant pathogens were isolated from 116 (11%) of 1, 099 consecutive specimens (80 from both conventional media and FAN bottles; 23 from FAN bottles only; 13 from conventional media only). Gram-positive cocci were more likely to be recovered from FAN bottles than from conventional media (p = 0.04). Contaminants were also more likely to have grown in FAN bottles (3%) than on conventional media (1%) (p = 0.04). The mean time to detection of significant pathogens was 20.9 h using FAN bottles as compared to 30. 9 h using conventional media (p = 0.0001). These results indicate that the BacT/Alert Microbial Detection System using FAN blood culture bottles improves the yield of clinically significant Gram-positive isolates from normally sterile body fluids with a reduced time to detection. PMID- 10794934 TI - Detection of PCR products of the ipaH gene from Shigella and enteroinvasive Escherichia coli by enzyme linked immunosorbent assay. AB - PCR techniques applied to diarrheal stools reliably diagnose Shigella and enteroinvasive Escherichia coli (EIEC) infections. Identification of PCR products using agarose gel electrophoresis (AGE) and hybridization with DNA probes has several shortcomings. Automated methods of identifying PCR products that process larger numbers of specimens can facilitate epidemiologic studies and standardize results. In this study, we used ELISA following PCR to detect ipaH gene sequences of Shigella and EIEC from 89 diarrheal stools. Results of ELISA were compared with AGE with and without DNA probe, and with culture. Two specimen preparation methods were compared as well: boiling/centrifugation, and purification with silicon dioxide (SiO(2)). Both PCR product-detection methods identified significantly more infections than did culture. PCR-ELISA detected significantly more infections than PCR-AGE when processed using SiO2 (P = 0.014). PCR-ELISA allows screening of larger numbers of specimens, automates test results, and avoids use of mutagenic reagents. PCR-ELISA is faster than PCR-AGE when testing large numbers of specimens, although not when testing small numbers of specimens. PMID- 10794935 TI - Detection of Bordetella pertussis in a clinical laboratory by culture, polymerase chain reaction, and direct fluorescent antibody staining; accuracy, and cost. AB - Control of Bordetella pertussis in the community is hampered by slow and insensitive diagnostic tests. We therefore examined the accuracy and cost of culture, direct fluorescent antibody (DFA) staining, and PCR in a routine clinical laboratory. Six hundred thirty seven nasopharyngeal swabs and aspirates in casamino acids transport medium were cultured, stained with polyclonal (Difco), and monoclonal (BL-5 and Accu-Mab) anti-B. pertussis reagents, and amplified by an IS481-specific PCR. PCR products were detected by a hybridization enzyme immunoassay kit (Gen-eti-k DEIA, DiaSorin), with confirmation by a second PCR in a separate laboratory. Sensitivities and specificities of culture, polyclonal DFA, monoclonal DFA, and PCR were 36 and 100%, 11.4 and 94.6%, 8.3 and 98. 4%, and 95.0 and 99.3%, respectively, with a prevalence of 15.7%. The DFA tests were the most economical, and the PCR cost was 31% higher than culture. This study suggests that with minor improvements in economy, pertussis PCR can be implemented in a clinical laboratory with marked improvement in diagnostic accuracy. PMID- 10794936 TI - Comparison of the MB/BacT and BACTEC MGIT 960 system for recovery of mycobacteria from clinical specimens. AB - A total of 543 specimens were cultured in parallel with the MB/BacT and BACTEC MGIT 960 systems and on the conventional solid media. Mycobacteria were identified from 95 (17.5%) specimens, including 63 (66.3%) Mycobacterium tuberculosis and 32 (33.7%) nontuberculous mycobacteria. The recovery rates for the MB/BacT, MGIT 960, and solid media were 91.6, 87.4, and 54.7%, respectively, for all mycobacteria; the recovery rates were 93.6, 88.9, and 63.4%, respectively, for M. tuberculosis complex alone, and 87.5, 84.4, and 37.5%, respectively, for all nontuberculous mycobacteria. The mean times to detection of all mycobacteria by individual systems were 13. 9, 8.7, 31.7 days for the MB/BacT, MGIT 960 and solid media, respectively, 13.9, 9.3, 32.9 days for M. tuberculosis alone, and 14. 1, 8.1, 27.2 days for all nontuberculous mycobacteria. The contamination rates of the MB/BacT and MGIT 960 were 10.2 and 5.4%, respectively. With regard to detection times and recovery rates, both automated systems are superior to the conventional media (all p < 0.005). As compared to the MB/BacT, the MGIT 960 detected mycobacterial growth more rapidly (p < 0.001), and had a lower contamination rate (p = 0.003); however, there was no statistically significant difference in recovery rates between these two systems. These results indicate that both MGIT 960 and MB/BacT systems are rapid, sensitive, and efficient methods for the recovery of mycobacteria from clinical specimens. PMID- 10794937 TI - Evaluation of the BACTEC MGIT 960 system for the recovery of mycobacteria. AB - We evaluated the BACTEC MGIT 960 system, which is a fully automated, non invasive, continuous monitoring system for the growth and detection of mycobacteria. Including respiratory and other specimens, 1,742 specimens were processed and inoculated into the BACTEC MGIT 960 and the BACTEC 460 TB Systems, as well as onto Lowenstein-Jensen (L-J) media. A total of 104 isolates of mycobacteria were recovered from all culture systems. This included Mycobacterium tuberculosis complex, Mycobacterium avium-intracellulare (MAI) complex and other mycobacteria (MOTT). The isolation rates for M. tuberculosis complex and MAI complex were comparable for the BACTEC 460 (54.8% and 13.5%) and the BACTEC MGIT 960 (51.9% and 13.5%). The overall isolation rate was less for BACTEC MGIT 960 (76.9%) which was due to lesser number of MOTT isolates recovered from this system. The mean times to detection (TTD) for all mycobacteria were 9.3 days for the BACTEC MGIT, 14.6 days for the BACTEC 460 and 21.6 days for L-J. A significant difference was observed when TTD was tested in relation to degree of positivity in smears, with the BACTEC MGIT maintaining the short TTD even with less number of bacilli in the smear. The contamination rates were, 6.4% for BACTEC MGIT, 2.9% for BACTEC 460 and 12.1% for L-J medium. The BACTEC MGIT 960 system shows performances comparable to the BACTEC 460 and seems to be a dependable, user friendly system. PMID- 10794938 TI - Performance of para-Pak Ultra ECOFIX compared with Para-Pak Ultra formalin/mercuric chloride-based polyvinyl alcohol for concentration and permanent stained smears of stool parasites. AB - ECOFIX is a mercury and formalin-free fecal preservative that can be used for concentration of stool specimens and preparation of permanently-stained slides. In this study, the standard two-vial ParaPak Ultra system was compared with ECOFIX Ultra for the detection of intestinal parasites. A total of 261 specimens in 92 sets (77 with 3 specimens, 15 with 2 specimens) were collected in ECOFIX, formalin, and low viscosity polyvinyl alcohol (LV-PVA). Concentrations were performed from ECOFIX using Hemo-De and saline and from formalin using ethyl acetate and formalin. To prepare permanently-stained smears, ECOSTAIN (a modification of Wheatley's trichrome stain) was used on ECOFIX material and Wheatley's trichrome stain was used on specimens preserved in PVA. A total of 157 protozoa and helminths were detected; 132 (84.1%) were recovered in formalin/PVA and 129 (82.2%) in ECOFIX. In permanently-stained smears, 139 protozoa were observed, 116 (83.5%) in PVA-preserved material and 117 (84.2%) in ECOFIX. Fecal concentration yielded 111 parasites (103 protozoa and 8 helminths), of which 98 (88.3%) were detected in formalin-fixed stool and 48 (43.2%) in ECOFIX. Significantly fewer ECOFIX-preserved concentrates were positive for Blastocystis hominis (35 versus 15, p-value <0.001) and Endolimax nana (19 versus 2, p-value <0.001). In conclusion, use of the ECOFIX Ultra collection device in combination with ECOSTAIN resulted in largely comparable recovery of enteric parasites to the conventional two-vial ParaPak Ultra system when both sedimentation-concentration and permanently stained smears were performed, and 2-3 specimens per patient were evaluated. PMID- 10794939 TI - Comparative in vitro bactericidal activity between cefepime and ceftazidime, alone and associated with amikacin, against carbapenem-resistant Pseudomonas aeruginosa strains. AB - Fifteen unique isolates of carbapenem-resistant Pseudomonas aeruginosa were selected for time-kill studies to assess the bactericidal activity of cefepime (CFP) and ceftazidime (CZD) (at 4 and 16 microg/mL), alone and associated with amikacin (AMK) (4 microg/mL). CFP proved more active than CZD (p < 0.05, Student's t test). Bactericidal activity after 24-h incubation was only achieved by the combination of CFP (16 microg/mL) plus AMK. The higher in vitro activity of cefepime over that of ceftazidime against imipenem-resistant P. aeruginosa strains highlights the differences of these drugs beyond Enterobacterspp. and Staphylococcus aureus. PMID- 10794940 TI - CPT-EIA assays for the detection of vancomycin resistant vanA and vanB genes in enterococci. AB - Cycling Probe Technology (CPT) was combined with a colorimetric enzyme-immuno assay (EIA) to develop two assays for the detection of vanA and vanB genes in vancomycin resistant enterococci (VRE). The CPT-EIA assay employs a gene-specific fluorescein labeled DNA-RNA-DNA probe that gets cleaved within the probe : target duplex. The cleaved DNA probe fragments dissociate from the target, making it available for further cycling. Following the separation of cleaved probe fragments, anti-fluorescein-horseradish peroxidase antibodies are used for the detection of uncleaved probes. The two CPT-EIA assays were used to screen a collection of 440 clinical isolates (Modrusan et al., 1999). All of the 154 VanA and 131 VanB isolates were correctly identified in the vanA and vanB CPT-EIA, respectively. The VanA and VanB isolates were differentiated from vancomycin sensitive enterococci (VSE) and also from the VanC isolates. In addition, an accurate VRE detection in the CPT-EIA assay was shown with cultures grown on eight different media. PMID- 10794941 TI - Comparison of oral immediate-release (IR) and extended-release (ER) metronidazole bactericidal activity against Bacteroides spp. using an in vitro model of infection. AB - We used an anaerobic infection model capable of simulating human serum pharmacokinetic parameters (C(max), C(min), half-life, and AUC) to compare the activity of oral immediate-release (IR) and extended-release (ER) MTZ formulations. Four oral regimens of MTZ plus a control regimen were simulated in the model: [i] (IR) 500 mg q8h, [ii] ER 750 mg q12h, [iii] ER 750 mg q24h, and [iv] ER 1500 mg q24h. Two clinical Bacteroides fragilis isolates (MICs 0.5, 4.0 microg/mL) and two non-fragilis Bacteroides isolates (MICs 0.5, 3 microg/mL) were studied. All four oral MTZ regimens exhibited rapid, bactericidal activity (> or =3-log(10) decrease from the starting inoculum) within 12 h against both fragilis and non-fragilis Bacteroides isolates. Overall, no appreciable difference in the rate of bacteria killing was noted among the four MTZ formulations against either B. fragilis isolates (P = 0.907, 0.737) or non-fragilis isolates (P = 0.809, 0.768). We conclude that ER MTZ dosed at 12 or 24-h intervals possesses equivalent bactericidal activity to standard IR MTZ dosed every eight hours against susceptible Bacteroides spp. PMID- 10794942 TI - Characterization of isolates associated with emerging resistance to quinupristin/dalfopristin (Synercid) during a worldwide clinical program. AB - Quinupristin/dalfopristin (Synercid) is an i.v. antibiotic active against serious Gram-positive infections. Its unique dual mode of action means that the potential for resistance development is expected to be low. To determine the incidence of in vitro emerging resistance in worldwide clinical studies, susceptibility to quinupristin/dalfopristin was measured for baseline pathogens and corresponding on- or post-study isolates from 880 evaluable patients. In comparative studies of community-acquired pneumonia, complicated skin and skin structure infections, and nosocomial pneumonia, the incidence of emerging resistance was low (1 of 453; 0.22%; 95% CI: 0. 01-1.4%). Resistance development occurred in only one pathogen (methicillin-resistant Staphylococcus aureus). In noncomparative studies, six instances (1.8% of 338 evaluable cases; 95% CI: 0.7 to 4.0%) of emerging resistance (all vancomycin-resistant Enterococcus faecium) were confirmed, accompanied by therapeutic failure in four cases. Molecular typing did not confirm the identity of one pair of strains. Overall, the incidence of emerging resistance to quinupristin/dalfopristin was low. PMID- 10794943 TI - Frequency of occurrence and antimicrobial susceptibility patterns for pathogens isolated from latin american patients with a diagnosis of pneumonia: results from the SENTRY antimicrobial surveillance program (1998). AB - The correct empiric choice of antimicrobial therapy in the treatment of pneumonia in hospitalized patients has established itself as a major therapeutic challenge to clinicians. Selection of an inappropriate antimicrobial agent could lead to increased rates of mortality and morbidity. Characteristics of pathogens responsible for this infection such as species prevalence, overall antimicrobial resistance rates, and mechanisms of detected resistance could serve as an invaluable resource to clinicians in making such therapeutic selections. This report addresses the aforementioned problems/needs by analysis of 712 strains isolated from the lower respiratory tract of patients hospitalized with a diagnosis of pneumonia in 10 Latin American medical centers in the SENTRY Antimicrobial Surveillance Program (1998). The four most frequently isolated pathogens (no/% of total) were: Pseudomonas aeruginosa (191/26.8%), Staphylococcus aureus (171/24.0%), Klebsiella spp. (86/12.1%), and Acinetobacter spp. (75/10.5%); representing nearly 75.0% of all isolates. More than 40 antimicrobial agents (23 reported) were tested against these isolates by reference broth microdilution methodology, and susceptibility profiles were established. The nonfermentative Gram-negative bacteria (P. aeruginosa and Acinetobacter spp.) exhibited high levels of resistance to the agents tested. Amikacin (77.5% susceptible) was the most active drug tested against P. aeruginosa 50.0% against the Acinetobacter spp. isolates. Based on published interpretive criteria, over 22.0% of the Klebsiella spp. and 12.5% of the Escherichia coli were classified as extended spectrum beta-lactamase (ESBL) producers. Of the cephalosporin class compounds tested against the Klebsiella spp. and E. coli isolates, cefepime demonstrated the highest rates of susceptibility (84.9% and 91.7%, respectively). This compound also fared well against the Enterobacter spp. isolates, inhibiting 88.2% of the isolates tested, many of which were resistant to ceftazidime and ceftriaxone. Resistance to oxacillin among the S. aureus isolates was nearly 50. 0%, with vancomycin, teicoplanin, and the streptogramin combination quinupristin/dalfopristin inhibiting all isolates. Several clusters of multiply resistant organisms were also observed, and further characterization by ribotyping and pulsed-field gel electrophoresis established possible patient-to-patient spread. The results of this study indicate that rates of resistance among respiratory tract pathogens continue to rise in Latin America, with specific concerns for the high prevalence of nonfermentative Gram-negative bacteria isolated, oxacillin resistance rates in S. aureus, and the epidemic dissemination of multiply-resistant strains in several medical centers. International surveillance programs (SENTRY) should assist in the control of escalating antimicrobial resistance in this geographic area. PMID- 10794944 TI - Chondritis attributable to Lactobacillus after ear piercing. AB - A diabetic patient was admitted with ear pain and purulent discharge after "high ear piercing" by a friend. After investigation she was found to have chondritis, complicated by an abscess. Wound cultures grew Lactobacillus species. Patient responded to parenteral antibiotics without suffering cosmetic deformity. Sterile technique, adherence to post-piercing ear hygiene, and avoiding piercing the auricular cartilage may prevent complications associated with ear piercing. Lactobacillus should be included in the differential of pathogens involved in post-piercing chondritis. PMID- 10794945 TI - Bacteremic cellulitis caused by Non-01, Non-0139 Vibrio cholerae: report of a case in a patient with hemochromatosis. AB - We report a case of bacteremia associated with hemorrhagic bullous skin lesions on the leg caused by non-01, non-0139 Vibrio cholerae in a 66-year-old man with hemochromatosis developed in an inland region. The organism was isolated from blood and bullae fluid. The patient was treated successfully with cefotaxime and doxycycline. This report emphasizes the potential of this organism to produce bacteremic cellulitis in people with underlying illness in the absence of usual epidemiological risk factors. PMID- 10794946 TI - DANMAP: monitoring antimicrobial resistance in Denmark. AB - The objectives of the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) are to monitor trends in resistance among bacteria from animals, food and humans, to monitor the consumption of antimicrobial agents and to determine the association between consumption and occurrence of resistance and to model transmission of resistance from animals to humans. DANMAP is based on the examination of representative bacterial isolates of animal and human pathogens, of zoonotic bacteria and of indicator bacteria. For food animals, both diseased and the healthy populations are studied. Isolates from all three reservoirs are examined for their susceptibility to a basic panel of antibiotics that includes representatives of the major classes of compounds, making comparison of resistance levels in the reservoirs possible. Isolates are stored in a strain collection and are available for further study. The data are stored in databases as MIC values or mm inhibition zones with all identifiers. A system for recording all use of the veterinary medicines, VETSTAT, is currently under implementation. For production animals, the consumption will be recorded for each herd, providing a basis for detailed modelling of the effect of consumption on resistance. PMID- 10794947 TI - The French antibiotic resistance monitoring programs. AB - Surveillance of antimicrobial resistance in bacteria from animal origin in France is organised by the French Agency for Food Safety (Agence Francaise de Securite Sanitaire des Aliments, AFSSA) through two types of networks. The first collects non-human zoonotic Salmonella strains in one centre (AFSSA, Paris) where they are tested for their antimicrobial susceptibility. The others, managed by AFSSA Lyon, deal with bovine pathogenic strains and are multicentric, that is they collecting antibiotic sensitivity and other data from the local public veterinary diagnostic laboratories. This requires standardisation of the methods used in each partner laboratory. Statistical analysis of any change in French resistance patterns can be monitored by these three networks either as a function of strain pathogenicity and/or of the ecological origin of the isolate. The system also encourages efficient collaboration between veterinarians and the laboratory. Such collaboration improves both the quality of routine antibiotic testing and understanding of the molecular mechanisms underlying resistance. PMID- 10794948 TI - Antibiotic resistance monitoring: the Spanish programme. The VAV Network. Red de Vigilancia de Resistencias Antibioticas en Bacterias de Origen Veterinario. AB - Antimicrobial resistance is a problem in modern public health and antimicrobial use and especially misuse, the most important selecting force for bacterial antibiotic resistance. As this resistance must be monitored we have designed the Spanish network 'Red de Vigilancia de Resistencias Antibioticas en Bacterias de Origen Veterinario'. This network covers the three critical points of veterinary responsibility, bacteria from sick animals, bacteria from healthy animals and bacteria from food animals. Key bacteria, antimicrobials and animal species have been defined for each of these groups along with laboratory methods for testing antimicrobial susceptibility and for data analysis and reporting. Surveillance of sick animals was first implemented using Escherichia coli as the sentinel bacterium. Surveillance of E. coli and Enterococcus faecium from healthy pigs was implemented in 1998. In July 1999, data collection on Salmonella spp. was initiated in poultry slaughterhouses. Additionally, the prevalence of vancomycin resistant E. faecium was also monitored. This network has specific topics of interest related to methods of determining resistance, analysis and reporting of data, methods of use for veterinary practitioners and collaboration with public health authorities. PMID- 10794949 TI - Antibiotic resistance monitoring in bacteria of animal origin: analysis of national monitoring programmes. AB - Methods of antibiotic resistance monitoring of bacteria from animals in 12 European countries were surveyed in 1998. Most laboratories used disk diffusion methods, usually expressing results qualitatively, although a few also expressed the results either as MICs or zone diameters. The number of antibiotics used ranged from 5 to 37 (mean 15) and the most common antibacterials were streptomycin, gentamicin, neomycin, ampicillin, tetracyclines, chloramphenicol and sulphonamides. Salmonellae were monitored by most centres but few-tested campylobacter regularly. Escherichia coli from a wide range of animal species were tested in nine countries. Enterococci were tested on a limited ad hoc basis in six countries. Staphylococci, streptococci and pasteurellae were also frequently monitored but the number of isolates tested showed wide variation. Overall the presentation of the results differed, but most programmes used disk diffusion, control strains and monitored similar bacteria. Thus, it may be possible to harmonise monitoring programmes within the EU. PMID- 10794950 TI - Monitoring of antibiotic resistance in bacteria of animal origin: epidemiological and microbiological methodologies. AB - The occurrence of antibiotic-resistant bacteria in food animals is a major public health threat. Information on the prevalence of resistance to specific drugs in both bacterial and animal species together with changes occurring over time, are necessary to understand the magnitude of the problem and to establish baselines for taking action. The aim of this paper is to define the minimum epidemiological and microbiological requirements for establishing a surveillance of antimicrobial resistance in bacteria of animal origin. Surveillance should involve different bacterial species, veterinary pathogens, zoonotic bacteria and commensal bacteria used as indicators. The collected data should be periodically updated and the reports distributed among practising veterinarians and regulatory authorities. These reports would be a useful tool for developing guidelines for the prudent use of antimicrobial agents in veterinary medicine and for action strategies. PMID- 10794951 TI - Detection of antibiotic resistance in vitro. AB - Testing the susceptibility of bacteria to antimicrobial agents is fundamental to the study of resistance. The qualitative methods that were originally devised evolved separately in different countries and as the need for more precise quantitative information became clear, different solutions to the problem emerged. Consequently, the methods now recommended vary widely between countries and achieving a consensus on standardisation has proved difficult. Sensitivity testing serves two purposes to provide meaningful results to the prescriber and to monitor changes in susceptibility of microbial populations. The diagnostic purpose is adequately served by the diversity of methods used, provided they are performed and controlled efficiently. Highly standardised methods are however needed for surveillance in the national and international context and to provide meaningful comparisons to be made between individual centres. These needs are best met by the use of agreed methods and interpretative criteria in sentinel laboratories. No sensitivity test method can provide the 'correct' answer all the time, if only because the 'correct' answer in therapeutic terms is not known with any certainty. Laboratories providing a sensitivity testing service can influence prescribing--and hence the development of resistance--in a number of different ways and the quest for standardisation at all costs should not obscure these important functions. PMID- 10794952 TI - Antibiotic treatment for animals: effect on bacterial population and dosage regimen optimisation. AB - Concern regarding antimicrobial resistance has led to proposals for the prudent use of antimicrobial agents. Whilst this is appropriate, it is not sufficient. This article proposes that dosage schedules should be developed to provide a basis for the rational use of antimicrobial drugs. This requires knowledge of resistance mechanisms and transfer, the biochemistry and structure of microorganisms and both the pharmacodynamics and pharmacokinetics of antimicrobial drugs. Dosage schedules should maintain concentrations at the site of infection in excess of MIC(90) for bacteriostatic drugs and bactericidal drugs acting primarily by time-dependent mechanisms whilst they should provide high AUIC and C(max)/minimum inhibitory concentration (MIC) values for agents acting mainly by concentration-dependent mechanisms. It is proposed that pharmacodynamic and population pharmacokinetic data should be integrated through use of the sigmoidal E(max) equation, together with mathematical modelling and appropriate statistical analyses, to take account of the natural variation in drug pharmacodynamics and pharmacokinetics. PMID- 10794953 TI - The control of microbial diseases in animals: alternatives to the use of antibiotics. AB - In animal husbandry the control and prevention of infectious diseases is of basic economic importance. The introduction of antibiotics to treat bacterial infections almost 50 years ago led to a dramatic improvement in animal production. The emergence of antibiotic resistant strains demonstrates that the treatment of bacterial infections can not rely on the use of antibiotics without some critical consideration. Special attention has been paid to the use of antibiotics in animals including antimicrobial growth promoters because these can contribute to the problems with antibiotic resistance in humans. This has strongly emphasized the need to introduce disease preventive methods. A theory and methods for the prevention of diseases is presented that is based upon the effect on the target animal population of microbial exposure, defence and immunity to infections and combinations of these. It is emphasized that antibiotics should be an integral part of other disease preventive methods and used only when other methods have failed. They should not be included in the first line of action. PMID- 10794954 TI - Ecological impact of antibiotic use in animals on different complex microflora: environment. AB - Different means of interaction between microecological systems in different animal hosts (including humans) and the environment may occur during the transfer of resistant bacteria and their resistance genes. Spread of resistance takes place in different ways with respect to clonal spread of resistance strains by the spread of wide host range plasmids and translocatable elements. Commensals in ecosystems have a special significance and a pronounced capacity for acquisition and transfer of resistance genes as with Enterococcus faecium and Escherichia coli in the gut flora or Pseudomonas spp. in aquatic environments. The route of transmission from animals to humans by meat products is well established. Other routes via water and food plants (vegetables) have been investigated less, although resistance genes transfer in aquatic environments as evidenced from sequence comparison of such genes (e.g. tetR, floR in Salmonella typhimurium DT104). Whether this is due to rare but important transfer events or whether there is a more frequent exchange in aquatic or terrestrial environments needs further elucidation. PMID- 10794955 TI - Epidemiology of resistance to antibiotics. Links between animals and humans. AB - An inevitable side effect of the use of antibiotics is the emergence and dissemination of resistant bacteria. Most retrospective and prospective studies show that after the introduction of an antibiotic not only the level of resistance of pathogenic bacteria, but also of commensal bacteria increases. Commensal bacteria constitute a reservior of resistance genes for (potentially) pathogenic bacteria. Their level of resistance is considered to be a good indicator for selection pressure by antibiotic use and for resistance problems to be expected in pathogens. Resistant commensal bacteria of food animals might contaminate, like zoonotic bacteria, meat (products) and so reach the intestinal tract of humans. Monitoring the prevalence of resistance in indicator bacteria such as faecal Escherichia coli and enterococci in different populations, animals, patients and healthy humans, makes it feasible to compare the prevalence of resistance and to detect transfer of resistant bacteria or resistance genes from animals to humans and vice versa. Only in countries that use or used avoparcin (a glycopeptide antibiotic, like vancomycin) as antimicrobial growth promoter (AMGP), is vancomycin resistance common in intestinal enterococci, not only in exposed animals, but also in the human population outside hospitals. Resistance genes against antibiotics, that are or have only been used in animals, i.e. nourseothricin, apramycin etc. were found soon after their introduction, not only in animal bacteria but also in the commensal flora of humans, in zoonotic pathogens like salmonellae, but also in strictly human pathogens, like shigellae. This makes it clear that not only clonal spread of resistant strains occurs, but also transfer of resistance genes between human and animal bacteria. Moreover, since the EU ban of avoparcin, a significant decrease has been observed in several European countries in the prevalence of vancomycin resistant enterococci in meat (products), in faecal samples of food animals and healthy humans, which underlines the role of antimicrobial usage in food animals in the selection of bacterial resistance and the transport of these resistances via the food chain to humans. To safeguard public health, the selection and dissemination of resistant bacteria from animals should be controlled. This can only be achieved by reducing the amounts of antibiotics used in animals. Discontinuing the practice of routinely adding AMGP to animal feeds would reduce the amounts of antibiotics used for animals in the EU by a minimum of 30% and in some member states even by 50%. PMID- 10794956 TI - Epidemiology and ecology of enterococci, with special reference to antibiotic resistant strains, in animals, humans and the environment. Example of an ongoing project within the European research programme. AB - The objectives of the present study are to generate knowledge of the ecology and epidemiology of enterococci in the food chain by studying the following: (1) the population structure (in measures of abundance, number of vancomycin resistant strains, antibiotic resistance patterns, diversity, and stability) among enterococcal populations in different geographical regions and in different links of the food chain (2) possible transmission of strains through the food chain and between hospital environments and the food chain (3) the association between vancomycin resistance and individual strains of enterococci and (4) the diversity of the drug resistance genes in enterococci. So far, 1578 samples have been collected from different countries within the EU (Sweden, Denmark, UK and Spain), and from different habitats (pig farms, carcasses in slaughter houses, soil, manure, water, sewage, and humans). Total and vancomycin resistant enterococcal populations in each sample have been enumerated and more than 12000 isolates have been characterised by phenotyping. Representative isolates are further species identified and characterised by genotyping and MIC determination and from antibiotic resistant isolates the resistance genes are characterised. PMID- 10794957 TI - Introduction. PMID- 10794958 TI - Leptin: an essential regulator of lipid metabolism. AB - This paper reviews the general mechanisms by which leptin acts as a regulator of lipid reserves through changes in food intake, energy expenditure and fuel selection, with an emphasis on its direct effects on cellular lipid metabolism. Briefly, when leptin levels increase, food consumption decreases via modulation of hypothalamic neuropeptides. As well, normal decreases in energy expenditures (e.g. with diurnal cycles or reduced caloric intake) do not occur. This is probably caused by an increase in mitochondrial proton leak mediated by leptin via increases in sympathetic nervous system stimulation and thyroid hormone release. The decrease in caloric input coupled with relatively higher energy expenditure, therefore, leads to negative energy balance. Leptin also changes the fuel source from which ATP is generated. Fuel preference switches from carbohydrate (glucose) to lipid (fatty acids). This effect arises through stimulation of triacylglycerol catabolism by leptin. In vitro studies show that leptin is a potent stimulator of lipolysis and fatty acid oxidation in adipocytes and other cell types. Consequently, leptin is also a regulator of cellular triacylglycerol content. Hormonal regulation of leptin, as well as its role in fasting and seasonal weight gain and energy expenditure are also briefly discussed. PMID- 10794959 TI - Living without oxygen: lessons from the freshwater turtle. AB - Freshwater turtles, and specifically, painted turtles, Chrysemys picta, are the most anoxia-tolerant air-breathing vertebrates. These animals can survive experimental anoxic submergences lasting up to 5 months at 3 degrees C. Two general integrative adaptations underlie this remarkable capacity. First is a profound reduction in energy metabolism to approximately 10% of the normoxic rate at the same temperature. This is a coordinated reduction of both ATP generating mechanisms and ATP consuming pathways of the cells. Second is a defense of acid base state in response to the extreme lactic acidosis that results from anaerobic glycolysis. Central to this defense is an exploitation of buffer reserves within the skeleton and, in particular, the turtle's shell, its most characteristic structure. Carbonates are released from bone and shell to enhance body fluid buffering of lactic acid and lactic acid moves into shell and bone where it is buffered and stored. The combination of slow metabolic rate and a large and responsive mineral reserve are key to this animal's extraordinary anaerobic capacity. PMID- 10794960 TI - Effects of capture on adrenal steroid and vasopressin concentrations in free ranging bottlenose dolphins (Tursiops truncatus). AB - Marine mammals are routinely caught in the wild in an effort to monitor their health. However, capture-associated stress could potentially bias various biochemical parameters used to monitor the health of these wild caught animals. Therefore, the effects of capture were quantified by measuring plasma adrenal steroids and arginine vasopressin (AVP) in free-ranging bottlenose dolphins (Tursiops truncatus) (n=31). Total capture and restraint times were also correlated to hormone concentrations to quantify the effects of capture. Significant, positive correlations between corticosterone and cortisol (R=0.752; P<0.0001), and between corticosterone and aldosterone (R=0.441; P=0.045) were demonstrated. Significant correlations between capture and restraint time and hormone levels were not observed. Animals restrained for less than 20 min exhibited hormone levels similar to those for animals restrained for more than 20 min. The positive correlations among the adrenal steroids suggest that release of these steroids was stimulated by adrenocorticotropin (ACTH). The lack of a correlation between cortisol and AVP indicates that AVP did not influence ACTH induced cortisol release in this situation. The study suggests that (1) a typical hypothalamic-pituitary-adrenal axis is present in these animals, and (2) the relatively short capture and restraint times did not induce a significant neuroendocrine stress response. PMID- 10794961 TI - Stress fever magnitude in laboratory-maintained California ground squirrels varies with season. AB - A previous study demonstrated that California ground squirrels (Spermophilus beecheyi) living in the natural environment had, independent of season, a significantly higher mean diurnal body temperature (T(b)) (39.6 degrees C) than either summer (37.5 degrees C) or winter (36.5 degrees C) laboratory maintained animals. Based upon the previous study it has been suggested that California ground squirrels living in the natural environment may have an elevated set-point for body temperature in a manner analogous to a stress fever response. The present study was conducted to determine if season and/or duration of laboratory open-field exposure influenced the magnitude of laboratory open-field stress fever. If stress fever was involved to some extent in the higher body temperature observed in animals from the natural environment, laboratory maintained animals should exhibit a lower magnitude stress fever during the summer months and a higher magnitude stress fever during the winter months. It was hypothesized that laboratory maintained animals would exhibit the same set-point for stress fever T(b) independent of season, and that the duration of open-field exposure would not influence the magnitude of stress fever. Adult California ground squirrels were acclimated to an ambient temperature of 20+/-1.0 degrees C under either LD 14:10 (summer) or LD 10:14 (winter) photoperiod conditions and individuals from both photoperiod conditions were exposed for periods of 2, 4, and 6 h to an open field arena. An analysis of the data with a two-factor ANOVA demonstrated that season (photoperiod) significantly influenced the magnitude of the stress fever response (1.1+/-0.1 degrees C for summer animals; 2.1+/-0.2 degrees C for winter animals) while there was no significant influence of open-field exposure duration on stress fever magnitude. These results demonstrate that although the set-point for body temperature in unstressed laboratory maintained California ground squirrels varies with season, the set-point for body temperature in open-field stressed animals does not vary with season. These data lend support to the hypothesis that something like stress fever may play some role in the higher body temperature observed in California ground squirrels living in the natural environment. PMID- 10794962 TI - The effect of ryanodine on isometric tension development in isolated ventricular trabeculae from Pacific mackerel (Scomber japonicus). AB - An isometric muscle preparation was used to study the inhibitory effect of ryanodine on contractile function in isolated ventricular trabeculae of the Pacific mackerel (Scomber japonicus). Ryanodine (an inhibitor of sarcoplasmic reticulum (SR) function) caused a 20% reduction in peak tension at 20 degrees C, but not 15 degrees C, over the range of frequencies (0.2-3.0 Hz) tested. This indicates that in the absence of a functional SR, the mackerel ventricle can maintain most of its contractile strength utilizing other modes of Ca(2+) delivery to the myofilaments. Ca(2+) flux through the sarcolemmal (SL) L-type Ca(2+)-channels is most likely the predominant pathway for Ca(2+) activation of the myofilaments, although reverse mode Na(+)/Ca(2+) exchange could potentially contribute to a significant extent. High levels of adrenergic stimulation overwhelmed the negative inotropy caused by ryanodine, returning tension to pre ryanodine levels, further suggesting that the mackerel ventricle can maintain contractile function without Ca(2+) contribution from the SR. These results are discussed within the context of what is known about SR Ca(2+) utilization in rainbow trout and tuna hearts. PMID- 10794963 TI - Chromosaponin I stimulates the sugar taste receptor cells of the blowfly, Phormia regina. AB - Chromosaponin I (CSI), a gamma-pyronyl-triterpenoid saponin isolated from pea and other leguminous plants, stimulates the growth of roots in a variety of plants. In the present work, we introduce CSI as a sugar taste substance for the blowfly, Phormia regina. The blowfly has taste chemosensilla on the labellum. The sensory receptor cells in the chemosensillum are highly specialized for the tastes of sugar, salt and water, respectively. Application of CSI induced the feeding response of blowflies including full proboscis extension. CSI also induced impulses of the sugar taste receptor cell in the LL-type sensillum. The optimum concentration of CSI in these responses was 0.1 mM which is much lower than that of sucrose. Based on the comparison of dose-response relationships, CSI is 100 times more effective than sucrose in stimulating the sugar taste receptor cells. CSI-induced impulses appeared after a significant latency compared with sucrose. As far as we know, this is the first report describing that a natural saponin induces sugar responses in insects. CSI is a unique saponin because of its bifunctional property in plants and insects. PMID- 10794964 TI - Involvement of tyrosine kinase and phosphatidylinositol 3-kinase in phagocytosis by ascidian hemocytes. AB - It has been proposed that protein tyrosine phosphorylation plays important roles in signal transduction in mammalian T- and B-cells and monocytes. During our investigations on the ascidian host defense system, we have shown that the monoclonal antibody A74 strongly inhibits both phagocytosis of sheep red blood cells (SRBCs) by hemocytes and hemocyte aggregation, and that the A74 antigen protein has two immunoreceptor tyrosine-based activation motifs and several other motifs that are thought to function in signal transduction in mammals. In this study, we found that the A74 antibody strongly inhibited phagocytosis by ascidian hemocytes of yeast cells, as strongly as that of SRBCs, but not that of latex beads. We also found that herbimycin A and an erbstatin analog, tyrosine kinase inhibitors, and wortmannin, a specific inhibitor for phosphatidylinositol 3 kinase (PI3-kinase), inhibited the phagocytosis of yeast cells. We investigated which hemocyte proteins were specifically tyrosine-phosphorylated during phagocytosis by ascidian hemocytes and found that a protein with a molecular mass of 100 kDa was specifically tyrosine-phosphorylated upon phagocytosis; its tyrosine phosphorylation was inhibited by the A74 antibody. These results strongly suggest that both tyrosine kinase and PI3-kinase play important roles in phagocytosis by ascidian hemocytes. PMID- 10794965 TI - Alpha(1)- and alpha(2)-adrenoceptor mediation in melanosome aggregation in cryptic patterning of Pleuronectes americanus. AB - The dark upper (ocular) surface of winter flounder (Pleuronectes americanus) possesses a chromatophore pattern, with cryptic dark bands and white spots, and a general background component, with melanophores under adrenergic neural control. In vitro responsiveness to the alpha-adrenoceptor agonists phenylephrine and clonidine and to the antagonists yohimbine and prazosin demonstrates that melanosome aggregation in this species is mediated through both alpha(1)- and alpha(2)-adrenoceptors, the alpha(2)-subtype being predominant in each pattern component. However, differences in the potency of agonists and antagonists indicate variability in the balance between the two receptors between pattern components. This paper demonstrates that it cannot be generalized that teleost melanophore alpha-adrenoceptors are universally of one subtype, and also that there is intraspecific variation in subtypes of these receptors associated with the flatfish cryptic patterning mechanism. PMID- 10794966 TI - Time course for cryoprotectant synthesis in the freeze-tolerant chorus frog, Pseudacris triseriata. AB - Increases in liver glycogen phosphorylase activity, along with inhibition of glycogen synthetase and phosphofructokinase-1, are associated with elevated cryoprotectant (glucose) levels during freezing in some freeze-tolerant anurans. In contrast, freeze-tolerant chorus frogs, Pseudacris triseriata, accumulate glucose during freezing but exhibit no increase in phosphorylase activity following 24-h freezing bouts. In the present study, chorus frogs were frozen for 5- and 30-min and 2- and 24-h durations. After freezing, glucose, glycogen, and glycogen phosphorylase and synthetase activities were measured in leg muscle and liver to determine if enzyme activities varied over shorter freezing durations, along with glucose accumulation. Liver and muscle glucose levels rose significantly (5-12-fold) during freezing. Glycogen showed no significant temporal variation in liver, but in muscle, glycogen was significantly elevated after 24 h of freezing relative to 5 and 30 min-frozen treatments. Hepatic phosphorylase a and total phosphorylase activities, as well as the percent of the enzyme in the active form, showed no significant temporal variation following freezing. Muscle phosphorylase a activity and percent active form increased significantly after 24 h of freezing, suggesting some enhancement of enzyme function following freezing in muscle. However, the significance of this enhanced activity is uncertain because of the concurrent increase in muscle glycogen with freezing. Neither glucose 6-phosphate independent (I) nor total glycogen synthetase activities were reduced in liver or muscle during freezing. Thus, chorus frogs displayed typical cryoprotectant accumulation compared with other freeze-tolerant anurans, but freezing did not significantly alter activities of hepatic enzymes associated with glycogen metabolism. PMID- 10794967 TI - Characterization of Na/K-ATPase in Macrobrachium rosenbergii and the effects of changing salinity on enzymatic activity. AB - A ouabain-sensitive Na/K-ATPase kinetic assay system based on the hydrolysis of ATP and the oxidation of NADH was adapted in order to characterize enzymatic activity in gills and examine the effects of changing salinity in Macrobrachium rosenbergii. Maximum inhibition by ouabain occurred at a concentration of 1.4 mM, and the K(m) of the reaction was 0.2 mM. In a first experiment, animals were acclimated to freshwater, 1/3 seawater, 2/3 seawater and full seawater for up to 1 week. Na/K-ATPase activity in front gills was 1. 62+/-0.19 micromol ADP/mg protein per h in freshwater, and was seen to increase slightly in 1/3 seawater (1.88+/-0.19 micromol ADP/mg protein per h) and 2/3 seawater (2.09+/-0.24 micromol ADP/mg protein per h), decreasing slightly in full seawater (1.92+/-0.43 micromol ADP/mg protein per h); however, differences were not significant. Back gills showed slightly higher levels, and a similar pattern of Na/K-ATPase activity. In a second experiment, animals were acclimated to 1/3 seawater and 2/3 seawater, and then transferred to freshwater. However, no changes in activity were seen, indicating that exposure to dilute media did not effect enzymatic activity. Whereas Na/K-ATPase is important in osmoregulatory function in marine euryhaline crustaceans, it may not play a significant role in adaptation in freshwater crustaceans that inhabit a more narrow range of salinities. PMID- 10794969 TI - Branchial chemoreceptors mediate ventilatory responses to hypercapnic acidosis in channel catfish. AB - The effects of hyperoxic hypercapnia on cardiovascular and ventilatory variables and blood gas and acid/base parameters were examined in conscious and anesthetized spontaneously breathing (ASB) channel catfish, Ictalurus punctatus. These separate experiments were designed to determine: (1) if channel catfish show a ventilatory response to hypercapnic acidosis when blood O(2) content is maintained in conscious animals; and (2) whether branchial receptors innervated by cranial nerves IX and X mediate this response. The combination of high O(2) and CO(2) tensions allowed the cardioventilatory effects of hypercapnic acidosis to be assessed independently of Root or Bohr mediated changes in blood O(2) content. In the absence of significant changes in dorsal or ventral aorta O(2) content, hyperoxic hypercapnia significantly stimulated ventilation, relative to hyperoxic exposure. Hypercapnic acidosis, however, had no significant effects on blood pressure or heart rate. Branchial denervation in ASB fish abolished the ventilatory response to hypercapnic acidosis. The results indicate that hypercapnic acidosis independently stimulates ventilation in channel catfish. This response is mediated by CO(2)/pH-sensitive branchial receptors innervated by cranial nerves IX and X. PMID- 10794968 TI - Gastrointestinal stability and absorption of insulin in suckling pigs. AB - Stability and absorption of orally administered fluorescein-isothiocyanate labeled insulin (FITC-insulin) in the gastrointestinal (GI) tract were investigated in newborn and 3-day-old pigs. The uptake of FITC-insulin by the intestinal epithelial cells was visualized using confocal laser scanning microscopy. Following oral administration, 3 h later 56 and 88% of orally administered fluorescence was found in the GI tract in newborn and 3-day-old piglets, respectively. Chromatographic analysis revealed that 15-37% of fluorescence recovered from the gastric and proximal intestinal contents was eluted in the void volume of a Sephadex G-25 column. It was also observed that oral administration of FITC-insulin at a dose of 100 nmol/kg body weight led to a significant decrease in blood glucose in newborn pigs (P<0. 05) but not in 3-day old pigs. Microscopic examination showed that FITC-insulin was taken up via the vesicular transport mechanism throughout the whole small intestine but the ileum appeared to be a preferred site for FITC-insulin transport in newborn pigs. In 3 day-old pigs, the uptake of FITC-insulin occurred only in the distal part of the small intestine. These findings suggest that milk-borne insulin may partially survive in the GI lumen and subsequently act on the gastrointestinal tract in suckling piglets, while GI absorption of milk-borne insulin is limited to newborn pigs. PMID- 10794970 TI - Corrigendum to "Nutrient, fatty acid, amino acid and mineral analysis of natural prey of the Hawaiian monk seal, Monachus schauinslandi". PMID- 10794971 TI - Alcohol sclerotherapy of hepatic cysts: its effect in relation to ethanol concentration. AB - While ultrasonically guided percutaneous aspiration and ethanol injection therapy has proved useful for non-neoplastic giant hepatic cysts that cause symptoms, the optimum quantity of ethanol injected into hepatic cysts has not been established. In eight patients with non-neoplastic giant hepatic cysts who were treated by ethanol injection, ethanol concentration in cystic fluid after treatment was estimated. Ethanol concentrations ranged from 5.9 to 47.6%. Hepatic cyst regressed almost completely in five cases; in all of which, the estimated ethanol concentrations were more than 40%. Our results suggest that it is desirable to drain cystic fluid as much as possible and to inject ethanol as little as possible, so that the estimated ethanol concentration in the cyst exceeds 40% for maximal effect and minimal side effects. PMID- 10794972 TI - Prevalence and risk factors of hepatitis C virus infection among Koreans in rural area of Korea. AB - In a Korean rural area with a high incidence of liver cancer, a molecular epidemiological study was conducted to estimate the prevalence of hepatitis C virus (HCV) infection and to determine the distribution of HCV genotype and transmission routes of HCV infection. The study population is those who volunteered to participate in a health survey in 1993 and were over 10 years of age. Those who were anti-HCV-positive and 20 of their spouses were re-examined in 1995 and tested for HCV genotype. A phylogenetic tree was constructed after sequencing analysis of core and envelope regions from the subjects with genotype 2a without a blood transfusion history. The age- and sex-standardized prevalence of anti-HCV was 5.52%. In a multivariate analysis, transfusion history was not associated with anti-HCV-positivity, but a history of acupuncture (adjusted odds ratio=2.2, 95% confidence interval=1.0, 4.7) and a history of surgical operation (adjusted odds ratio=2.0, 95% confidence interval=1.0, 4.1) were associated. The prevalence of HCV genotype 2a was the highest, and genotype 1b was less frequent. The phylogenetic tree showed strong homology among our samples with Japanese HCV strains. The present study suggests that there is a highly endemic area of HCV infection in Korea and that this endemicity is probably associated with a parenteral source of HCV infection other than blood transfusion. The study also suggests that some of the HCV infection was spread through non-sterilized or non disposable acupuncture needles in this HCV endemic area. PMID- 10794973 TI - Two-year follow-up study after treatment with lamivudine for chronic hepatitis B: seven cases reported. AB - 2 times baseline) and returned to seropositive for HBV DNA and HBeAg after lamivudine cessation. One of these five patients died of liver failure 3 months after treatment. However, in two of five patients whose alanine aminotransferase (ALT) had rebounded, HBV DNA became undetectable, and the ALT levels markedly decreased 2 years after the end of therapy. Since the disappearance of HBV DNA and stabilization of the ALT level were observed within two patients by 2 years after cessation of treatment, the patients whose ALT had rebounded should be followed up for a long-term period. To confirm the effect of lamivudine, long term follow-up in many patients is necessary. PMID- 10794974 TI - A case of acute hepatitis A with marked hemophagocytosis in bone marrow. AB - A previously well 18-year-old female was referred to our hospital because of abnormalities of blood biochemistry and slight jaundice. Because serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were elevated more than 6000 IU/L, the patient was suspected to have acute viral hepatitis. The platelet count on admission was 9.7x10(4)/ul, which was decreased from the initial value of 21x10(4)/ul for 3 days. The coagulation tests revealed marked elevation of D-dimmer, fibrinogen degradation products and thrombin-antithrombin III complex suggesting increase in fibrinolysis. Serum levels of high density lipoprotein cholesterol and ferritin were markedly decreased and increased, respectively. The bone marrow smears revealed proliferation of mature histiocytes ingesting platelets and erythrocytes, these pathological findings were consistent with those of hemophagocytic syndrome (HPS). In addition, anti-hepatitis A IgM antibody in the serum and hepatitis A virus (HAV) RNA in the stool were positive. Therefore, the patient was diagnosed as having acute hepatitis A with probable HPS. Since a fulminant clinical course was suspected, glucocorticoid pulse therapy was started immediately 7 days after onset and a favorable clinical outcome resulted. PMID- 10794975 TI - Contribution of the portal flow on hepatic tissue perfusion increases in reduced hepatic vascular bed. AB - Background: A hepatic artery-portal vein reciprocal response and hepatic hemodynamics have well been investigated under normal condition, but not under pathologic condition with decreased vascular bed. This study was designed to determine the hemodynamic changes in the hepatic blood flow, tissue perfusion and interrelationship between portal venous flow (PVF) and hepatic arterial flow (HAF) after inflow interruption in the various size of hepatic vascular bed. Methods: Anesthetized dogs were used to measure PVF and HAF using a transit time flow meter and hepatic tissue flow (HTF) using a laser Doppler flow meter before and after portal venous (PVO) or hepatic arterial occlusion (HAO) under various range of portal triad occlusion (PTO). Results: The ratio of HAF/TLF (total liver flow) was 38+/-14% under the basal condition. This ratio did not change under the 30% PTO where there was a similar decrease in PVF and HAF, but reduced to 25+/ 12% under the 70% PTO where there was more selective reduction in HAF than PVF. Although a reciprocal HAF increase was observed under any conditions after PVO, the TLF and HTF decreases after PVO were largest under the 70% PTO with the highest PVF/TLF ratio. On the other hand, there was no reciprocal PVF increase in any conditions after HAO, and the TLF and HTF decreases after HAO were minimal under the 70% PTO with the lowest HAF/TLF ratio. Conclusions: With decreasing hepatic vascular bed, dependency of the remnant hepatic hemodynamics and tissue perfusion on the portal blood flow increased. These findings suggest that an integrity of portal venous flow becomes crucial in the remnant hepatic tissue perfusion after extensive hepatic resection. PMID- 10794976 TI - Experimental study on liver regeneration after simultaneous partial hepatectomy and pancreatectomy. AB - In this study, we performed hepatectomy and pancreatectomy to assess the physiological contribution of the pancreas, especially in terms of endocrine function to hepatic regeneration. Group 1 Wistar rats underwent 70% hepatectomy and group 2 rats underwent 70% hepatectomy plus 50% pancreatectomy. The time course assessment of liver regeneration rates obtained by Fishback's formula demonstrated a difference in rates between the two groups as early as day 3 or day 7 after surgery. Since levels of both PCNA-positive cells and serum transforming growth factor-alpha (TGF-alpha) were significantly higher in the hepatectomy only group, we could prove the difference of liver regeneration between the two groups. We have concluded that pancreatectomy retards the liver regeneration initiation processes occurring from 24 h to 3 days after evisceration. Glucagon-insulin molar ratios most significantly differed between the two groups 3 days after evisceration in the present study. This result was due to increased glucagon level of group 2 at day 3 after evisceration. Our findings suggest that 50% partial pancreatectomy inhibits the rate of hepatic regeneration, thereby altering the supply of pancreatic hormones, especially glucagon. PMID- 10794977 TI - Regional differences in cerebral glucose metabolism in cirrhotic patients with subclinical hepatic encephalopathy using positron emission tomography. AB - Although it is recognized that abnormalities of glucose metabolism occur substantially throughout the brain in cirrhotic patients with subclinical hepatic encephalopathy (SHE), little attention has been paid to these abnormalities in situ due to technical difficulties. The aim of this study is to clarify the regional differences of cerebral glucose metabolism (CMRglu) in patients with or without SHE using positron emission tomography (PET). Fifteen patients with posthepatitic cirrhosis and 13 control subjects underwent PET imaging of the brain using 2-[18F]fluoro-2-deoxy-D-glucose. Neurological tests included the digit symbol and block design subtests from the Wechsler Adult Intelligence Scale. Auditory brain stem reaction and electroencephalogram were also used in diagnosis of SHE. Regions were defined as frontal, parietal, occipital, basal ganglia, and the white matter. The CMRglu value in the grey matter of SHE is lower than that of control, particularly in the frontal and temporal regions and basal ganglia (P<0.05, respectively). The CMRglu values in the grey matter of non SHE and control are almost the same, excepting the basal ganglia. The CMRglu value in the basal ganglia of non-SHE is higher than that of control and SHE. The CMRglu value in the grey matter of SHE is depressed as compared to both non-SHE and control. These data suggest that the abnormalities of cerebral glucose metabolism may be a contributing factor in SHE. PMID- 10794978 TI - [Endonasal dacryocystorhinostomy (DCR) with protected drill]. AB - PURPOSE: To analyze the interest of protected drills to perform endoscopic endonasal DCR. MATERIAL AND METHODS: [corrected] Fifty patients suffering from chronic epiphora were operated successively. None of those patients had had prior nasal surgery. The same surgeons (BF and ER) did the surgical procedure for all the cases. The technical steps were: endoscopic visualization of the middle meatus and evaluation of the anatomic variations of the nose; identification of the lacrymal bag with a 1 mm light in the canaliculus; unciformectomy; the maxillary bone was drilled with a 3.2 mm bit (straightshot Xomed), protected and irrigated; the lacrymal bag was opened by cissors and the flap was pushed in the ethmoid and coagulated. RESULTS: The suction-irrigation system was efficient to clean the surgical site, with a good endoscopic control. The protected bit didn't damage the 4-mm optics and the septal wall mucosa, even in narrow noses. The staightshot bit (Xomed) was tough enough to drill the maxillary bone in only 68 % of the surgical cases. In 28 % the drilling was unsatisfactory and in 4 % it was inefficient. The functional results at 6 month were good in 85 %. There was no difference between normal and narrow noses. CONCLUSION: The concept of protected bits perfectly matches the endoscopic endonasal DCR. The visual comfort and security of the surgical procedure are increased. Short term results are equivalent to external DCR, but are rather inefficient and expensive. PMID- 10794979 TI - [Malignant tumors of the eye and adnexa in Congo-Kinshasa]. AB - PURPOSE: To determine the frequency and to study anatomic and clinical features of malignant tumors of the eye and adnexa. METHODS: A retrospective (1962-1990) and prospective (1990-1992) study of 164 medical records of patients with histopathologic confirmation of malignant tumors of the eye and adnexa. All the patients received a conventional ophthalmological examination and underwent surgical biopsy. Histopathologic examinations were performed for a large part at the Department of Ophthalmology, University of Iowa and for a small part at the Department of Histopathology, University Hospital of Kinshasa. RESULTS: There were 99 (60%) male and 65 (40%) female for a sex ratio of 1.5: 1. The mean age of patients was 24.6 +/- 21.4 years. Epibulbar (35%) and intraocular (33%) tumors were the most frequent, followed by orbital tumors (20%). Epidermoid carcinoma and retinoblastoma were the most common histologic forms, representing respectively 33.5% and 31.7% of all malignant tumors of the eye and adnexa. They occurred especially in adults between 20 and 60 years for the first and in children below 5 years for the second. Metastatic tumors accounted for 9.7% of all cancers of the eye and adnexa. Kaposi's angiosarcoma was encountered in 3.6% of cases and in 3% it was associated with AIDS. Burkitt's lymphoma was seen in 3% of cases, basal cell carcinoma as well as in situ carcinoma in 2.4% of cases. Malignant lymphoma and adenocarcinoma both accounted for 1.8% of all cases of cancers of the eye and adnexa while adenoid cystic carcinoma, liposarcoma as well as rhabdomyosarcoma accounted for 1.2% of all cases. Other types of tumors were seen in very small proportion. CONCLUSION: Our results were similar to those of other studies in Africa and were different from those of European and American studies. PMID- 10794980 TI - [Retreatment after PRK for low and medium myopia. Results and study of contrast sensitivity]. AB - PURPOSE: To examine the results of retreatment with eximer laser photorefractive keratectomy (PRK) on refraction, visual acuity, haze, and contrast sensitivity. PATIENTS AND METHODS: Excimer laser photorefractive keratectomy was performed in 18 patients, once for one eye and twice for the other eye due to undercorrection or regression. Outcome was compared between the two eyes. RESULTS: Uncorrected visual acuity was above 20/40 in 94.4% of the retreated eyes. There was no statistical difference between the two eyes for haze and contrast sensitivity. CONCLUSION: Retreatment with laser photorefractive keratectomy is a safe procedure and should be performed when regression or undercorrection occurs after the first laser excimer treatment. PMID- 10794981 TI - [Exploration of retro-chiasmatic visual pathways in human albinism]. AB - PURPOSE: Several research studies have explored the abnormal crossing of the retinogeniculate and geniculocortical optic pathways in human albinos. This prospective study has dealt with visual evoked potentials (VEPs) of human subjects to identify the percentage of albinos with asymmetric VEPs. PATIENTS AND METHODS: A series of 16 albino patients ranging in age from 6 to 37 years were examined. They had measurable visual acuity, with or without nystagmus. Diffusion of flash stimuli not allowing selective study of the two visual pathways (direct and crossed), two stimulation patterns were used for VEP recordings: monocular full open field then hemi-field stimulation to isolate the activity of each visual pathway. ANALYSIS: In the normally pigmented subject, fibers derived from the nasal half of the retina of each eye decussate at the chiasma, while temporal retinal fibers are uncrossed and project to the ipsilateral hemisphere. In albinos, the majority of temporal retinal fibers subserving the nasal field (from fixation to an eccentricity of about 20 degrees ) anomalously cross with the nasal retinal fibers. Therefore with monocular stimulation, the evoked visual response should be obtained only in the contralateral hemisphere. The asymmetry, morphology and latency for the first major positive peak and the amplitude of the VEP were examined and compared with the normal population. CONCLUSION: We managed to demonstrate the characteristic VEP asymmetry only in 3 out of the 16 patients. The results presented herein lead to question the absolute validity of VEP abnormality in diagnosis of albinism for clinical purposes. PMID- 10794982 TI - [Therapeutic photokeratectomy for the treatment of band keratopathy]. AB - PURPOSE: This retrospective study was designed to investigate the therapeutic potential of phototherapeutic keratectomy (PTK) for the treatment of band keratopathy. PATIENTS AND METHODS: PTK was performed with the Excimed UV 200, (Summit Technology, Inc) on 27 eyes of 22 patients and with the Nidek EC5000 on 11 eyes of 10 patients with band keratopathy. Mean patient age was 57 years. The mean follow-up period was 19 months (ranged from 4 to 54 months). When possible, the change in best corrected visual acuity and spherical equivalent was evaluated at 6, 12, 18 and 24 months. RESULTS: Almost 90% of patients with smooth bands achieved visual improvement and 85% of patients with rough bands improved ocular discomfort. The mean hyperopic shift caused by tissue ablation was less than +2.0D after one year, for smooth bands. CONCLUSION: Excimer laser PTK is a safe and effective outpatient treatment and should be used as initial treatment for band keratopathy. PMID- 10794983 TI - [Effect of scleral indentation on the corneal topography and the axial length after retinal detachment surgery. A prospective study in 30 cases]. AB - INTRODUCTION: Scleral indentation is an essential time in conventional retinal detachment surgery. It enables re-establishing retinal contact and to counterbalance vitreal tractions. It can be circular, segmental or radial. Silicone materials are often used. MATERIALS: In a prospective study, we followed up 30 eyes of patients who underwent retinal detachment surgery. We analyzed changes in corneal surface using differential map of the corneal (induced astigmatism, meridian axial deviation) and change in the axial length of the globe. RESULTS AND DISCUSSION: Follow-up examination showed corneal astigmatism: 2.62 diopter (D) at one week, 2.37D at one month and 1.80D at 3 months. Corneal axis meridians changed: 25 degrees at one week and 18.6 degrees at 2 months. Axial length was also modified. We observed a lengthening: 1.7mm during the first week which remained unchanged at 1.63 after 2 and 3 months. These changes were studied according to the different techniques used in our study and were compared with results reported in the literature. CONCLUSION: We think that conventional retinal detachment surgery causes modifications in refractis (induced astigmatism and axial lengthening). PMID- 10794984 TI - [Comparison of visual results and quality of vision between two multifocal intraocular lenses. Multifocal silicone and bifocal PMMA]. AB - PURPOSE: To compare visual results and quality of vision of two multifocal intraocular lenses (IOL): SA 40 N silicone zonal-progressive multifocal IOL and P 359 TUV PMMA bifocal IOL. METHODS: 46 eyes were operated on for cataract. 24 IOL SA 40 N (group 1) and 22 IOL P 359 (group 2) were implanted. Evaluation of visual results was performed between the first and the third month postoperatively. The quality of vision was explored in both groups by using two tests of contrast sensitivity, an automated visual field and a questionnaire dealing with patient satisfaction and presence of halos and glare. RESULTS: Postoperatively, a myopic shift was noted with a statistical significant difference in the group 2 (p<0.05). The predictability was good in both groups (mean refractive error<1 D). Mean corrected distance visual acuity was similar in-groups 1 and 2 (0.6). Mean distance-corrected near visual acuity was statistically better in the group 2 (p<0.05). We noted in both groups a loss of contrast sensitivity statistically more important in the group 2 (p<0.001). Where the automated perimetry is concerned, no statistical differences were encountered. 35% of patients (group 1) and 29% (group 2) were very satisfied with the results of their surgery. Halos and glare were noted in 8% (group 1) and 9% (group 2) of the patients. CONCLUSION: The new concept of multifocality is increasing in practice of cataract surgery because of the reduction of spectacle dependency and the high level of patient satisfaction. However, the occurrence of halos and glare suggest a limitation of these indications of multifocal IOL, particularly for night driving patients. PMID- 10794985 TI - [Slit-lamp laser photocoagulation with a quadraspheric contact lens for the treatment of retinopathy of prematurity]. AB - PURPOSE: To present our experience of slit-lamp laser photocoagulation with a pediatric quadraspheric contact lens for the treatment of retinopathy of prematurity (ROP). MATERIAL AND METHODS: 8 eyes of 4 premature babies with threshold ROP were consecutively treated under general anesthesia by means of blue-green argon laser photocoagulation delivered through a table mounted slit lamp and a wide field contact lens specially designed for infants. Three of the treated eyes had a zone I disease, 2 a posterior zone II disease and 3 a zone II disease. Mean follow-up was 13 months. RESULTS: Early complete regression of the neovascular proliferations and plus disease was observed in all the treated eyes (100%) without apparent sequela. In only one eye that had zone I disease a mild and small peripheral vitreous hemorrhage was observed. Neither cataract nor posterior synechia was observed. A discrete and transient badly hypothermia occurred only in one case. COMMENTS AND CONCLUSIONS: This study shows the efficacy and precision of slit-lamp argon laser photocoagulation through a quadraspheric contact lens in ROP, with a very low rate of complications, and with a magnificent and comfortable visualization of all the structures to be treated. Each retinal burn was easily and very precisely placed from the ridge to the ora serrata in 360 with only a slight to moderate tilting of the wide field lens and without the need of scleral depression. PMID- 10794986 TI - [Temporo-spheno-orbital meningioma. An unusual cause of exophthalmos. Report of one case]. AB - We report the case of a 48-year-old woman with no prior medical history who developed headache and unilateral exophthalmos for 6 months. Clinical and radiographic findings detailed the intracranial extension of a temporo-spheno orbital meningioma. Histological diagnosis was established after curative neurosurgery. This case recalls the characteristic features of temporo-spheno orbital meningiomas. PMID- 10794987 TI - [Xeroderma pigmentosum and its ocular manifestations. The first case in Cameroon]. AB - We report a case of xeroderma pigmentosum in a 9-year-old back Cameroonian boy. The diagnosis was based on typical clinical presentation of the disease: cutaneous atrophy, hypepigmented macules, and areas of depigmentation on sun exposed regions of the skin. Multiple tumoral lesions were localized on the head. Ocular findings were also present: conjunctival hyperemia, peripheral corneal opacification. Excision of the tumors and potoprotection was proposed for this patient. The role of tribal black African marriage traditions in disease transmission is discussed. PMID- 10794989 TI - [Pain in ophthalmology: toward better management]. PMID- 10794988 TI - [Acquired ocular toxoplasmosis (panuveitis) after liver transplantation]. AB - We present one case of acquired Toxoplasma gondii retinochoroiditis after livrer transplantation. Three weeks after receiving liver transplant from a seropositive donor for Toxoplasma gondii, the fundus examination showed a vitreal haze and an edematous retinal lesion with retinal hemorrhages. Routine screening tests for uveitis were performed. The serum toxoplasmosis titers revealed a seroconversion for toxoplasmosis (pre-transplantation tests were negative). The PCR assay on aqueous humor resulted in an amplified product identical to toxoplasmosis. This immunocompromised patient likely received an infected hepatic transplant. The patient's ocular inflammation cleared on 1-month pyrimethamine-sulfadiazine medication. Four months later, the patient developed a retinal detachment with massive proliferative vitreoretinopathy. PMID- 10794990 TI - [Multidisciplinary approach to pain and institution of analgesic treatment]. AB - Pain is a complex sensory response to tissue damage but also mental functioning. Patients with chronic pain require an interdisciplinary model of care after a careful evaluation and understanding of pathophysiology. Therapy may include medications, nerve blocks and behavioural interventions. PMID- 10794991 TI - [Pain and corneal pathology]. AB - Since the development of photorefractive keratectomy, ophtalmologists are concerned by the mechanisms of corneal pain in order to prevent and reduce it. Corneal epithelium and anterior stroma are widely innervated by nasociliary fibers form ophtalmologic nerve (V). Receptors react to a chemical, mechanical and thermal stimulation by an acute and precisely located pain. This pain is developed by release of inflammatory mediators and neuro transmitters. Pain is different regarding the pathology and the treatment: photorefractive keratectomy, Acanthamoeba keratitis, Mooren's ulcer. Topical non steroidal antiinflammatory drugs have a specific role in controlling corneal pain. PMID- 10794992 TI - [Pain and zona ophthalmica]. AB - Pain before and during Herpes Zoster is well known. A new antiviral therapy is suggested. Post-herpetic neuralgia remains more difficult to treat. PMID- 10794993 TI - [Clinical significance of pain after retinal detachment surgery]. PMID- 10794995 TI - [ [In Process Citation] PMID- 10794994 TI - [Per- and postoperative pain]. AB - We have made a huge amount of progress over the last decade, but we must continue to clear a way through the undergrowth. Per- and postoperative pain is typical of acute pain and is characterized by an excess of nociception due to tissue injury, visceral distention or disease. It contrasts with chronic pain which has been going on for 3 to 6 months. PATHWAYS OF PAIN: We know that the spinothalamic cord is involved in nociceptive activity. Two pathways are of great interest. One ends in a nucleus of the hypothalamus and the second in the amygdala. Both are involved in affective and emotional activity. Treatment of postoperative pain requires first that one be aware that pain is a reality and secondly that it must be assessed with various scales. Only then can the therapeutic strategy set about using the different methods available. Strategy must be continuously assessed to improve its efficacy. Different kinds of treatment can be used during the per- and postoperative period including enteral (paracetamol, nonsteroidal analgesic and antiinflammatory drugs (NSAIDs)) or parenteral (paracetamol, NSAIDs, morphine or similar) analgesic drugs. Local anesthesia seems however to be more effective. Several factors are involved in per- and postoperative pain. Its treatment is based on comprehensive team work in which each person represents a link in the therapeutic chain. For the future, we can expect to see the appearance of new drugs, processes or associations able to prolong postoperative analgesia. PMID- 10794997 TI - [ [In Process Citation] PMID- 10794996 TI - [Choroidal melanoma. An anatomic-clinical case report]. AB - A 78-year-old-male-patient had his left eye affected by a 10 mm thick choroidal malignant melanoma. The size of the tumor did not allowed to keep the eye, and no conservative treatment was used for therapy. Enucleation was performed and histopathological study of the eye showed a mushroom-shaped and mixed-cell choroidal melanoma, devoid of transcleral extension, but harboring intrascleral infiltration. A period of 2 year-follow-up without metastasis was observed since the removal of the tumoral eye. PMID- 10794998 TI - [Doppler ultrasonography of carotid atheromatous lesions]. AB - Carotid stenosis is the most frequently factor involved in stroke. The diagnosis of these lesions in routinely based on ultrasonic examination which allows to detect the carotid stenoses which were essentially atheromatous. B-mode evaluates the fine echostructure of plaques and Doppler helps to quantify the degree of stenosis using direct and indirect hemodynamic criteria. Echography with the help of color Doppler imaging identifies the site of stenosis in order to determine the degree of stenosis as a percentage of normal carotid area. Moreover echography is an essential method to appreciate the surface as well as the echostructure of the carotid lesion which have to be taking into account for a well-adapted therapy. The maximal velocities determined at the tight site of the lesion, the spectral changes downstream the stenosis and the hemodynamic changes observed upstream and downsteam of the carotid stenosis allow a precise quantification of this stenosis. The information obtained using sonographic methods (including 2 direct hemodynamic criteria, 2 indirect hemodynamic criteria and, if possible, a B-mode assessment of the area) are essential to appreciate the neurological risk in these patients. PMID- 10794999 TI - [Ultrasound contrast agents]. AB - Ultrasound contrast agents are introducing a new vision to ultrasonography. The increase in the Doppler signal intensity improves difficult examinations. Technical failures in the study of transcranial, liver and renal vessel might be rescued with the injection of these new compounds. The introduction of specific image sequences such as harmonic imaging and pulse inversion imaging allows detection of parenchymal enhancement in liver and renal examinations. The injection of ultrasound contrast agents also provides dynamic quantitative information to develop functional imaging. However, it is mandatory to understand the interaction with the ultrasound beam in order to optimise their use and define the adequate ultrasound sequence. PMID- 10795000 TI - [Echocardiographic diagnosis of left intraventricular thrombi: contribution of high frequency probes]. AB - Two-dimensional echocardiography is the reference technique for the diagnosis of left ventricular thrombi. However, it has some limitations due to poor imaging of the apex; location of many thrombi. Apical resolution improvement is possible using a 5 MHz ultrasonic transducer, which may identify 3.5MHz ultrasonic transducer false positive results. In a series of 53 patients with left ventricular dysfunction, we detected 11 thrombi. Using the 5MHz ultrasonic transducer as a reference the sensitivity of the 3.5MHz ultrasonic transducer was 100%, and much greater than that of the 2.5MHz ultrasonic transducer (55%), which was associated with 4 false positive results. The increased sensitivity associated with transducers of higher frequency was however limited by the echogenicity of patients, all the more since we used a medium instead of a short focalisation. Due to its therapeutic consequences, the detection of left ventricular thrombi must be enhanced using 5 MHz ultrasonic transducer, all the more in echogenic patients. PMID- 10795001 TI - [Internet use for telemedecine: fetal medicine applications]. AB - PURPOSE: Assessment of the transmission of still and moving images via the Internet for teleconsultations in fetal medicine. MATERIAL: and methods: Implementation fo two PC systems linked via the internet for the transmission of fixed ultrasound, radiography and scanner images, as well as moving ultrasound images. The assessment took into account was based upon the compression factor. RESULTS: The commutative telephone network transmitted ultrasound and radiographic images by e-mail without difficulty, rapidly (3min 46s for 4Mo files, after compression), inexpensively, and well. The transmission of video ultrasound files using the same procedure was also of good quality, but and exorbitant amount of time was required to send and receive the files. A more advanced configuration could improve the transmission time only sligthly, to 2h30 min 19s for 15s sequences. Live tranmission of video ultrasound sequences by the internet cannot be considered with anything less than a connection including ISDN lines, the minimum to provide the necessary bandwith. CONCLUSION: The Internet is a valuable tool to allow expert consultation in foetal medicine of live still images and recorded moving images. PMID- 10795002 TI - [Variability of ultrasonography velocity assessment of the carotid arteries]. AB - PURPOSE: To assess the intraobserver variability of echoDoppler sonographic measurement of various carotid arteries parameters. MATERIALS AND METHODS: Repeated bilateral carotid examinations were performed in ten healthy subjects, using three color-coded duplex ultrasound systems (two Acuson 128 XP, one ATL HDI 3000) and different linear probes. Each examination included two successive sets of measurements. RESULTS: The resistive index values decreased from the proximal common carotid artery to the distal common carotid artery (p <10(-7)). According to the set of measurements and to the equipments considered, the range of mean absolute differences of the systolic peak velocity (PSV), the end diastolic velocity (EDV) values, the ICA/CCA PSV and PSV/EDV velocity ratios and the resistive index (RI) values were 8-16cm/s, 3-6cm/s, 0.08-0,15 and 0. 16-0.26, and 0.02-0.06 respectively. Standard deviations of the proportional differences of the RI and PSV measurements averaged respectively 6% and 15% and were always reduced by repeating the measurements. The use of similar ultrasound machines was not associated with a reduced variability. CONCLUSION: The substantial variability of Doppler measurements in carotid arteries, even with the use of the same operator and similar machines could be reduced using "angle independent" parameters. PMID- 10795003 TI - [Carotid restenosis after endarterectomy. Treatment by angioplasty and endoprosthesis]. AB - Endovascular treatment of carotid arteries stenoses is an acceptable therapeutic option to surgery in some selected patients. In this case report a patient successfully underwent carotid stenting in the presence of a severe restenosis following a surgical endarterectomy. PMID- 10795004 TI - [Simultaneous management of coronary and cerebral vascular lesions in patients with multiple atheromatous lesions]. AB - This case report highlights the complementarity of noninvasive imaging modalities in the diagnosis of extracranial cerebral arteries lesions. The need for a strategy in the treatment of patients with combined lesions is discussed. PMID- 10795005 TI - [Bladder candidosis after renal transplantation: contribution of ultrasonography]. AB - Candidosis of the ureter and the bladder is an infrequent complication after renal transplantation. A case occurring early after kidney transplantation is reported in a patient presenting with candiduria. Ultrasound examination detected several fungal bezoars, presenting as a proliferating, avascular lesions, located within the ureter and the bladder. It allowed to demonstrate the rapid extension of lesions, in spite of a medical treatment, leading to a successful endoscopic removal of fungus balls. PMID- 10795006 TI - [Contributions of endosonography in ovarian neoplasms: Report of 3 cases]. AB - Ovarian investigations are necessary to answer to two questions Intraoperative explorations play a main part for that decision. Endoscopic sonography could contribute to distinguish benign from malignant lesions. We present three cases of endoscopic sonography of ovarian disease. In each case, endoscopic explorations induced to change usual surgical procedure. The authors suggest that endoscopic sonographic becomes an intraoperative step of ovarian surgery. PMID- 10795007 TI - [What is your diagnosis? Doppler color of bilateral testicular lymphoma]. PMID- 10795008 TI - [B-flow ultrasonographic imaging of circulating blood]. AB - B-Flow is a new imaging technique which uses General Electric's Digitally Encoded Ultrasound technology to provide direct visualization of blood echoes in grey scale imaging. This allows simultaneous imaging of blood flow and vessel walls with a high frame rate and high spatial resolution without information on velocity and direction. This promising technique which improves early detection of vascular disease could modify our routine practice in a near future. PMID- 10795009 TI - [In Process Citation] PMID- 10795010 TI - [Positron emission tomography to study central pain integration]. AB - The study of pain integration, in vivo, within the human brain has been largely improved by the functional neuro-imaging techniques available for about 10 years. Positron Emission Tomography (PET), complemented by laser evoked potentials (LEP) and functional Magnetic Resonance Imaging (fMRI) can nowadays generate maps of physiological or neuropathic pain-related brain activity. LEP and fMRI complement PET by their better temporal resolution and the possibility of individual subject analyze. Recent advances in our knowledge of pain mechanisms concern physiological acute pain, neuropathic pain and investigation of analgesic mechanisms. The sixteen studies using PET have demonstrated pain-related activations in thalamus, insula/SII, anterior cingulate and posterior parietal cortices Activity in right pre-frontal and posterior parietal cortices, anterior cingulate and thalami can be modulated by attention (hypnosis, chronic pain, diversion, selective attention to pain) and probably subserve attentional processes rather than pain analysis. Responses in insula/SII cortex presumably subserve discriminative aspects of pain perception while SI cortex is particularly involved in particular aspects of pain discrimination (movement, contact.) In patients, neuropathic pain, angina and atypical facial pain result in PET abnormalities whose significance remain obscure but which are localized in thalamus and anterior cingulate cortices suggesting their distribution is not random while discriminative responses remain detectable in insula/SII. Drug or stimulation induced analgesia are associated with normalization of basal thalamic abnormalities associated with many chronic pains. The need to investigate the significance of these responses, their neuro-chemical correlates (PET), their time course, the individual strategies by which they have been generated by correlating PET data with LEP and fMRI results, are the challenges that remain to be addressed in the next few years by physicians and researchers. To advance our knowledge of the mechanisms generating both abnormal pain and analgesia (drugs and surgical techniques) in patients is the main motivation of such anexciting challenge. PMID- 10795011 TI - [Amyotrophic lateral sclerosis: a role for enteroviruses?]. AB - Amyotrophic lateral sclerosis (ALS) is the most frequent motor neuron disease. Although it is a clinically well defined disease, its etiology remains unknown. Among, various hypotheses, the putative role of enteroviruses has been recently suggested by the detection of enteroviral sequences in neurons of spinal cord samples from ALS patients. However, there is a lack of consensus on the role of EV in ALS. In the present paper, we summerized the pathogenic role of these viruses, analyzed the discrepancy between different studies and speculated on the possible role of enteroviruses in ALS. PMID- 10795012 TI - [Interest of a computerized ALS database in the diagnosis and follow-up of patients with ALS]. AB - The aim of the study is to present a computerized database of the Neurology department of Limoges University Hospital and the main results obtained from data of 340 patients suffering from amyotrophic lateral sclerosis (ALS), diagnosed between 1984 and 1997. It is a user friendly and can be accessed by all neurologists at any level of computer knowledge. This database is modular (6 modules) and flexible according to need. The software used, Access 7, is an open relation database, which allows export of data to statistical or other compatible software. One of the reasons, which led to the elaboration of this database was to develop a means of collecting data in an analyzable manner for therapeutic trials. During these trials, a great number of data can be collected during each clinic visit for the evaluation of the degree of impairment, disability, and handicap. We present part of the data from the patients followed, some of whom were treated with riluzole, the current reference molecule for ALS. PMID- 10795014 TI - [Lower motor neuron disease and signs of dysimmunity]. AB - Twenty-two patients (12 men, 10 women, age range 16 to 60) affected with an adult onset, sporadic, lower motor neuron disease were studied. Motor weakness was associated with a severe muscular atrophy but never in a peripheral nerve distribution. Weakness predominated in the proximal parts of the limbs in 3 cases, in distal parts in 10 cases involving predominantly the upper limbs in 10. It was diffuse in all four limbs in six cases and was monomelic in the last 2 two others. Reflexes were generally lost in weak muscles. Electrodiagnostic findings consisted of pure motor axonal features, subtle sensory involvement was present in 3 cases with an IgM monoclonal gammopathy, in only one case the neurological syndrome was associated with a lymphoproliferative disorder despite complete investigations. All patients had dysimmune biological features (MGUS or anti-GM1 antibodies). We studied SMN gene in 12 patients and found no deletion. 16 patients were treated with IVIg and five improved but in 2 cases the improvement was transcient and lasted less than six months. Intravenous cyclophosphamide (1g/m(2) repeated monthly during 6 to 9 months) was used in six patients and three improved. Among these three patients two received also plasma exchanges on two days before the infusion. In all three patients, muscle weakness gradually deteriorated in the months following the end of the treatment. Nor the weakness pattern nor the type of biological marker was predictive of a good response to treatment. Lower motor neuron diseases appear to be much less sensitive to treatment than multifocal motor neuropathy with conduction block. However, treatment with IVIg or cyclophosphamide must be considered in the most severe forms or in case of a young onset. PMID- 10795013 TI - [What's new in primary lateral sclerosis?]. AB - Primary lateral sclerosis as a nosological entity distinct from amyotrophic lateral sclerosis has been the subject of controversy since it was first described in the nineteenth century. Primary lateral sclerosis has been defined as a rare, non-hereditary disease characterized by highly progressive spinobulbar spasticity, related to the exclusive loss of precentral pyramidal neurons, with secondary pyramidal tract degeneration and preservation of anterior horn motor neurons. We carried out a study in nine patients with a diagnosis of primary lateral sclerosis. Our clinical, electrophysiological and pathological investigations provide evidence that the disease has a heterogeneous clinical presentation and that degeneration is not restricted to the central motor system but also affects the lower motor neuron. In view of this similarity with amyotrophic lateral sclerosis, primary lateral sclerosis may represent a slowly progressive syndrome closely related to motor neuron disease and amyotrophic lateral sclerosis. PMID- 10795016 TI - [Bilateral medial medullary infarction]. AB - We report a case of a 42-year old man with a bilateral medial medullary stroke (MMS) with favorable outcome. First described by Spiller in 1908, the MMS accounts for less than 0,5% of all cerebral infarcts. It may be unilateral or more rarely bilateral, and may often be the consequence of the atherosclerosis. The clinical features of MMS classically associate contralateral hemiparesis and lemniscal hypoesthesia accompanied by ipsilateral lingual palsy and sometimes oculomotor disturbances (upbeat nystagmus). With the advent of magnetic resonance imaging, some restricted or unusual clinical manifestations can be attributed to this localization. The benign form of MMS seems much more common than MMS with poor prognosis. PMID- 10795015 TI - [Akinetic mutism and progressive supranuclear palsy-like syndrome after the shunt of an obstructive hydrocephalus. Successful treatment with bromocriptine: 2 cases]. AB - Two cases of obstructive hydrocephalus who suffered multiple shunt failures and shunt revisions are presented. The patients developed after the neurochirurgical treatment a clinical syndrome of akinetic mutism followed by a Progressive Supranuclear Palsy-like (PSP) syndrome. The akinetic mutism and the PSP-like syndrome were remarkably improved with bromocriptine. PMID- 10795017 TI - [Unusual complication of rendu-osler-weber disease: paramedian bulbar syndrome]. AB - We report a case of arteriovenous malformation of the brainstem, revealed by a progressive right pyramidal syndrome and an atrophy of the left hemi-tongue in a man presenting a Rendu-Osler-Weber disease. After embolization the clinical course was stationary. PMID- 10795018 TI - [Retroclival hematoma in a patient taking oral anticoagulants]. AB - Retroclival hematoma associated with anticoagulation. We report a case of retroclival hematoma associated with anticoagulant therapy in a 78-year-old woman, who presented with an isolated left external ophthalmoplegia. The role of the anticoagulant therapy is discussed. There was a spontaneous neurological resolution. PMID- 10795020 TI - [Brainstem tuberculomas: two cases]. AB - We report two cases of brainstem tuberculomas with favorable outcome after medical treatment alone. Tuberculomas should be suspected in all cases presenting space-occupying lesions of the brainstem. PMID- 10795019 TI - [Thoracic myelopathy revealing a chronic ossificans arachnoiditis of the thoraco lumbar spinal cord]. AB - We report a case of chronic spinal, ossificans, thoraco-lumbar arachnoiditis, associated with an arachnoidal cyst at the level of TH7, and revealed by a rapidly progressive thoracic myelopathy. Two years ago, he had presented with a spinal intrathecal haemorrhage of unknown etiology. There was no evidence of spinal traumatism, arterial hypertension or coagulation disorder. The finding of several small cysts and angiomas in the liver of this patient lead us to suspect an associated spinal vascular malformation, not detected on the spinal magnetic resonance imaging. A partial neurologic improvement was observed after laminectomy and cysto-peritoneal derivation. PMID- 10795021 TI - [Bilateral phrenic involvement disclosing Parsonage Turner syndrome]. PMID- 10795022 TI - [Right hemiparesis lymphocytic meningitis and sicca syndrome with a favorable outcome in a 28-years-old man]. PMID- 10795023 TI - [Neuropsychological evaluation: a useful clinical tool for neurodegenerative diseases]. PMID- 10795024 TI - [Discovery of the tendon reflexes Erb-Westphal-1875]. PMID- 10795026 TI - Internal Fixation of the Cervical Spine: Current Indications and Techniques. AB - Numerous techniques for the internal fixation of the cervical spine have been developed in recent years. The indications for surgery must be strictly defined before the optimal type of implant can be selected on the basis of the advantages and potential risks. Wiring techniques are still commonly used for posterior stabilization. Anterior fusion can be accomplished without internal fixation in most cases. The halo vest is still widely used for the conservative management of cervical fractures and for postoperative external immobilization. Posteriorly, traditional wiring methods are still used, as well as newer techniques, such as C1-C2 screw fixation, occipitocervical plate fixation, and lateral-mass lower cervical plating. Anteriorly, meticulous Smith-Robinson surgical approach and grafting techniques are essential. Further studies are needed to define the exact indications for various new techniques, such as anterior odontoid screw fixation and anterior plating. The surgeon must choose an appropriate device on the basis of the mechanism of injury, the pathoanatomy of the lesion, and familiarity with the device, keeping in mind the goals of internal fixation-stabilization, reduction and maintenance of alignment, early rehabilitation, and perhaps enhancement of fusion and avoidance of the need to use an external halo vest. PMID- 10795025 TI - Ankle Pain in Children: Diagnostic Evaluation and Clinical Decision Making. AB - Ankle pain in children can be caused by traumatic injuries to bone, ligament, or tendon or by nontraumatic conditions, such as congenital and developmental anomalies, infections and other inflammatory disorders, neural compression, metabolic derangements, and neoplasia. Evaluation of children with this complaint should include a focused history and an anatomically oriented physical examination. Depending on the findings, further diagnostic workup and laboratory evaluation should be done. Appropriate treatment-whether casting, surgery, antibiotic therapy, or a combination thereof-can then be selected on a rational basis. PMID- 10795027 TI - Femoral-Shaft Fractures in Children and Adolescents. AB - Femoral-shaft fractures are frequent in children and adolescents. Fortunately, most unite rapidly without significant complications or sequelae. Treatment options include spica casting, traction, external fixation, compression plating, and flexible or locked intramedullary nailing, each of which has advantages and dis-advantages. Treatment is determined primarily on the basis of the age of the patient and the presence of associated injuries, but psychological, economic, and environmental factors also must be considered. The appropriate treatment can be determined only by careful consideration of all factors and close consultation with the family. Meticulous attention to technical details will help decrease the incidence of complications from both nonoperative and operative treatment. PMID- 10795028 TI - The Diabetic Foot. AB - Management of foot problems in the patient with diabetes mellitus requires attention to each system affected by the disease. Appropriate treatment of common clinical problems affecting the foot in diabetic patients, such as ulcerations and fractures, depends on a thorough understanding of the pathophysiology of the disease. Treatment of neuropathy is directed at pressure relief and prevention of deformity. Infection is addressed with antibiotics, debridement, and improvement of the vascularity and oxygenation of the tissues. Amputation should be viewed, not as evidence of treatment failure, but as a reconstructive procedure, the goal of which is to regain energy-efficient ambulation. The orthopaedic surgeon can play a critical role in the team approach to the care of the diabetic patient with foot problems. PMID- 10795029 TI - Management of the Mangled Hand and Forearm. AB - In this article the authors describe the methods and procedures that have been used to help patients regain hand and forearm function after mangling injuries. Assessment of the pathophysiologic condition, careful inventory of the injured structures, and early aggressive wound excision and reconstruction, followed by expert rehabilitation, are advocated. The importance of vascular restoration, stable skeletal fixation, and provision of adequate skin cover are stressed. In addition to describing their treatment approach, the authors emphasize the need for the surgeon to be well acquainted with each phase of the treatment process. PMID- 10795030 TI - Osteochondritis Dissecans of the Knee: Treatment of Juvenile and Adult Forms. AB - Osteochondritis dissecans (OCD) and juvenile osteochondritis dissecans (JOCD) are distinct entities that require different management. Although both conditions result from stress fractures of the subchondral bone, JOCD has a much better prog nosis; treated conservatively, 50% of cases will heal, probably providing a normal knee during adult life. In contrast, OCD often is followed by the early onset of degenerative arthritis. The treatment of JOCD and OCD, whether nonoperative or operative, should be based on the principles of fracture treatment. Unfortunately, surgical correction of either of these conditions is unlikely to succeed unless the joint surface is perfectly restored. PMID- 10795031 TI - Infected Total Hip Arthroplasty: Diagnosis and Treatment. AB - The diagnosis of a deep infection complicating total hip arthroplasty is not difficult in most patients. When the diagnosis is not evident on the basis of the medical history, physical examination, routine blood work, and plain radiographs, indium-111-labeled leukocyte scintigraphy can be diagnostic. New immunologic techniques may allow differentiation of aseptic loosening from septic loosening of a painful total hip arthroplasty. Once the diagnosis of a deep infection about a total hip arthroplasty has been established, there are several treatment options. Oral antimicrobial therapy combining rifampin with a fluoroquinolone may prove to be an attractive alternative to surgical intervention in the treatment of some staphy-lococcal infections. If the causal microorganism is considered to be less virulent and does not elaborate glycocalyx, a one-stage procedure for reconstructing the hip with a cemented total hip arthroplasty incorporating antibiotics that are cidal to the microorganism has been successful in as many as 90% of patients. If the causal microorganism is considered to be virulent, a two stage procedure with a prolonged interval between the Girdlestone resection arthroplasty and the second-stage reconstructive procedure is the treatment of choice. PMID- 10795032 TI - Pedicle-Screw Fixation in the Lumbar Spine. AB - This article provides a perspective on the development of pedicle-screw fixation in the lumbar spine, the biomechanics of its application, the possible complications, and the scientific evidence that supports specific applications in selected disorders. The overall goal is to objectify the debate currently surrounding the use of these devices. Lumbar-pedicle fixation devices are currently considered class III med-ical devices. According to the Food and Drug Administration, such devices are investigational or experimental, have not been proved safe and effective, and may potentially pose a risk to patients. PMID- 10795033 TI - Genu Valgum in Children: Diagnostic and Therapeutic Alternatives. AB - Genu valgum is a common orthopaedic problem in children. The vast majority of cases are physiologic variants, which resolve normally. However, there are pathologic entities due to both focal and systemic processes in which the deformity often progresses and usually requires treatment. Differentiating between the two forms is facilitated by a thorough understanding of the natural history of the development of the tibiofemoral angle in children. In this review, an approach to the evaluation and diagnosis of genu valgum is presented, and therapeutic alternatives are discussed. PMID- 10795034 TI - The Dislocated Knee. AB - Knee dislocation, although relatively rare, may be the result of high-or low velocity injuries. Well established is the need for urgent diagnosis and treatment to avoid vascular complications and amputation. The initial evaluation should include objective assessment of arterial circulation by means of Doppler pressure measurements; the finding of any asymmetric pressure warrants an arteriogram. Late arterial occlusion may occur, which mandates careful serial reexamination in all patients, including those with initially symmetric pressure. Injury to the peroneal nerve is also common, and the recovery of neurologic function is unpredictable. An operative approach for the young and otherwise healthy patient is outlined. In the absence of definitive clinical studies, the timing and extent of the repair/reconstruction and the optimum rehabilitation still remain uncertain. Therefore, individual patient management must be dictated by circumstances such as instability, swelling, activity level, and the risk of postoperative joint stiffness. PMID- 10795036 TI - Ankle Arthroscopy: II. Indications and Results. AB - Diagnostic indications for the use of ankle arthroscopy include unexplained pain, swelling, stiffness, instability, hemarthrosis, and locking or popping, as well as a negative workup in a patient with significant ankle symptoms unresponsive to conservative care. Therapeutic indications include injuries of the articular cartilage and soft tissue, bone impingement, debridement of soft-tissue lesions, synovectomy and loose-body removal, arthrofibrosis, ankle fractures, and osteochondral defects. Ankle arthroscopy can also be used in ankle-stabilization procedures and arthrodesis, as well as for irrigation and debridement of septic arthritis. An algorithm has been developed to facilitate selection of the appropriate treatment for a patient with chronic ankle pain of unknown etiology. When used for the appropriate indications, ankle arthroscopy appears to give good results. PMID- 10795035 TI - Ankle Arthroscopy: I. Technique and Complications. AB - Arthroscopic surgery of the ankle allows the direct visualization of all intra articular structures of the ankle without an arthrotomy or malleolar osteotomy. Technological advances and a thorough understanding of anatomy have resulted in an improved ability to perform diagnostic and operative arthroscopy of the ankle. The decreased morbidity and faster recovery times make it an appealing technique compared with open arthrotomy. A keen understanding of the anatomy of the foot and ankle is critical to safe performance of arthroscopic procedures and prevention of complications. PMID- 10795037 TI - Closed Tibial-Shaft Fractures: Which Ones Benefit From Surgical Treatment? AB - Closed tibial-shaft fractures can usually be managed effectively with cast or brace immobilization if acceptable alignment is maintained and cyclic loading (weight-bearing) is initiated early. However, certain tibial fractures are at greater risk for nonunion or malunion and merit consideration for early operative stabilization. Among the tibial fracture characteristics that warrant fixation are instability, metaphyseal-diaphyseal location, significant limb edema, and the need for repeated realignment procedures. Deleterious patient-specific factors, such as obesity, poor compliance, and health conditions favoring immediate function, should also be considered. Absolute criteria for stabilization include coronal angulation exceeding 5 degrees, sagittal angulation greater than 10 degrees, rotation greater than 5 degrees, shortening exceeding 1 cm, displacement greater than 50%, and severe comminution (loss of 50% or more of cortical circumferential continuity). Relative indications for fixation include an inability to bear weight, distal or oblique fractures, prominent edema, and patient-specific considerations necessitating early function. When tibial stabilization is preferable, the authors believe that closed locked intramedullary nailing is the treatment of choice. PMID- 10795038 TI - Halo Skeletal Fixation: Techniques of Application and Prevention of Complications. AB - The halo skeletal fixator provides the most rigid cervical immobilization of all orthoses. However, complications such as pin loosening and infection are common. Appreciation of local anatomy and adherence to established application guidelines should minimize pin-related problems. A relatively safe zone for anterior pin placement is located 1 cm above the orbital rim and superior to the lateral two thirds of the orbit. Posterior pin-site locations are less critical; positioning on the posterolateral aspect of the skull, diagonal to the contralateral anterior pins, is generally desirable. Pins should enter the skull perpendicular to the cortex, with the ring or crown sitting below the widest portion of the skull and passing about 1 cm above the helix of the ear. Pins are inserted at a torque of 8 in-lb and retightened once to 8 in-lb at 48 hours. A loose pin can be retightened to 8 in-lb if resistance is met; otherwise, a loose pin should be replaced at a nearby site. Superficially infected pins are managed with local pin care and oral antibiotics. Persistent or severe infections require pin replacement to a nearby site, parenteral antibiotic therapy, and incision and drainage as needed. In ability to maintain acceptable cervical reduction with a halo fixator is an indication for alternative treatment, such as internal fixation or traction. PMID- 10795039 TI - Ligament Healing: Current Knowledge and Clinical Applications. AB - The treatment of ligament injuries, particularly knee-ligament injuries, has occupied a substantial portion of the orthopaedic literature for several decades. It remains unclear, however, what orthopaedic surgeons can do to optimize the recovery of patients with ligament problems. In this review, the reasons for this lack of clarity are proposed, and the current state of laboratory knowledge about the response of isolated and multiple ligament injuries to various treatment modalities is reviewed for the ligaments that have been studied thus far (all of which are in the knee). In general, it appears that ligaments heal with scar tissue similar to that involved in skin-wound healing. The early controlled motion of stable (or surgically stabilized) joints appears to improve ligament scar behavior, but no treatment identified to date stimulates true ligament regeneration. PMID- 10795040 TI - Fingertip Injuries: Evaluation and Treatment. AB - The primary goal of treatment of an injury to the fingertip is a painless fingertip with durable and sensate skin. Knowledge of fingertip anatomy and the available techniques of treatment is essential. For injuries with soft-tissue loss and no exposed bone, healing by secondary intention or skin grafting is the method of choice. When bone is exposed and sufficient nail matrix remains to provide a stable and adherent nail plate, coverage with a local advancement flap should be considered. If the angle of amputation does not permit local flap coverage, a regional flap (cross-finger or thenar) may be indicated. If the amputation is more proximal or if the patient is not a candidate for a regional flap because of advanced age, osteoarthritis, or other systemic condition, shortening with primary closure is preferred. Composite reattachment of the amputated tip may be successful in young children. The outcome of nail-bed injuries is most dependent on the severity of injury to the germinal matrix. PMID- 10795041 TI - Current Status of Anticoagulation Therapy After Total Hip and Total Knee Arthroplasty. AB - Postoperative venous thromboembolism in the pelvis and lower extremities is a potentially life-threatening complication in patients undergoing elective total hip and total knee arthroplasty. Numerous pharmacologic methods of prophylaxis have been used in the past with varying degrees of success. Warfarin has been proved effective as a prophylactic agent after total hip arthroplasty but has been less efficacious after total knee arthroplasty. The low-molecular-weight heparins have recently been approved for prophylaxis after total hip and total knee arthro-plasty and are an acceptable alternative to warfarin. This new class of drugs appears to have the advantage of predictable subcutaneous bioavailability, which allows less frequent administration and laboratory monitoring and offers a decrease in the occurrence of side effects. PMID- 10795042 TI - Osteochondral Lesions of the Talar Dome. AB - Osteochondral lesions of the talar dome are relatively common causes of ankle pain and disability. Trauma is the most common cause, but ischemic necrosis, en docrine disorders, and genetic factors may have etiologic significance. Medial lesions are usually located posteriorly on the dome of the talus, whereas lateral lesions are most frequently located anteriorly. Although the staging system described by Berndt and Harty remains popular, it may not accurately reflect the integrity of the articular cartilage. Small lesions of the talar dome may be present despite a normal appearance on plain radiography. Bone scintigraphy may show increased radionuclide uptake in the talar dome. Magnetic resonance imaging is also sensitive for identifying intraosseous abnormalities in the talus and has the added benefit of revealing other types of soft-tissue lesions not visible on routine radiographic studies. Computed tomography remains the imaging technique of choice when delineation of a bone fragment is desired. Nonoperative management of osteochondral lesions, including restricted weight-bearing and/or immobilization, is recommended unless a loose fragment is clearly present. Surgical options include drilling (usually reserved for intact lesions), debridement of the lesion with curettage or abrasion of the bone bed, internal fixation of the fragment, and bone grafting. Recent technical advances allow these procedures to be performed arthroscopically, with potential reduction of surgical trauma, length of hospital stay, and complication rates. PMID- 10795043 TI - The Painful Knee: Arthroscopy or MR Imaging? AB - While neither arthroscopy nor magnetic resonance (MR) imaging is perfect, both can delineate pathologic changes in the knee with reasonable accuracy. The greatest advantage of MR imaging is that it is noninvasive and can be used to detect pathologic changes both inside and outside the synovial cavity. Arthroscopy has the distinct advantage of allowing definitive treatment at the time of diagnosis in most cases, but carries with it the potential risks associated with any invasive diagnostic technique. Both modalities are expensive, and their judicious use is therefore dictated, especially in this era of cost containment. The decision to use one or both studies is best made by the orthopaedic surgeon. PMID- 10795045 TI - Periarticular Fractures After Total Knee Arthroplasty: Principles of Management. AB - Periarticular fractures about total knee replacements are sustained by 0.3% to 2% of patients who have undergone knee arthroplasty. The patient with such a fracture is usually a woman in her seventh decade who has osteoporosis and may also have rheumatoid arthritis that is being treated with corticosteroids. The treatment of such fractures is aimed at restoring the patient's functional status to the pre-fracture level. Accomplishing this requires healing of the fracture and retention of a mobile and painless prosthesis in correct alignment. These goals are often difficult to achieve because there is little experience with these uncommon fractures, the healing environment is suboptimal, and knee arthroplasties have a low tolerance for any resulting alteration in alignment. In general, nondisplaced fractures are treated nonoperatively, and displaced fractures require open reduction, rigid internal fixation, and bone grafting. If the prosthesis is loose, or if rigid fixation cannot be obtained, component revision is the treatment of choice. PMID- 10795044 TI - Neuropathic Arthropathy: Review of Current Knowledge. AB - Neuropathic arthropathy is a chronic, progressive degenerative disorder affecting one or more peripheral or vertebral articulations, which develops as the result of a disturbance in the normal sensory (pain or proprioceptive) innervation of joints. Diabetes, syphilis, and syringomelia are the most commonly associated clinical entities. When neuropathic arthropathy is suspected, careful clinical evaluation should be performed to identify an underlying neurologic disorder. Patient education, joint protection, and early recognition of fractures are the most important general management principles. Surgery can be considered in cases of advanced joint destruction when there is significant disability. PMID- 10795046 TI - Acute and Chronic Instability of the Elbow. AB - Elbow instability is not a single entity, but rather a spectrum of injuries, both acute and chronic. While acute instability is usually quite easily diagnosed and treated, in virtually all instances the chronic condition is much more problematic. Nonoperative treatment and early motion are recommended for acute injuries. An associated fracture decreases the likelihood of a good result. A thorough knowledge of the normal and pathologic anatomy, as well as a clear understanding of the osseous and soft-tissue reconstructive options, is essential, particularly for the proper management of recurrent and chronic elbow instability. Even for patients with the latter, however, reasonable treatment options are available. PMID- 10795047 TI - Popliteal Cysts: Historical Background and Current Knowledge. AB - Popliteal cysts were first described in 1840 by Adams, but it is from Baker's writing in 1877 that we derive the commonly used eponymic term "Baker's cyst." Associated intra-articular lesions are very common with popliteal cysts. Ultra sonography, arthrography, and magnetic resonance imaging have all proved useful in distinguishing popliteal cysts from other cysts and from soft-tissue tumors about the knee, as well as in identifying coexisting intra-articular lesions. Cysts in pediatric patients are generally self-limited and should be treated conservatively. In the adult population, treatment is primarily nonsurgical. Arthroscopic evaluation is indicated if an intra-articular lesion is causing mechanical symptoms or if there is no response to appropriate conservative treatment, such as use of nonsteroidal anti-inflammatory drugs and compression sleeves. Surgical excision is reserved for cases in which this approach has been unsuccessful. PMID- 10795048 TI - An Approach to Work-Related Disorders of the Upper Extremity. AB - Management of work-related musculoskeletal disorders has been frustrating for the orthopaedist. The so-called cumulative-trauma disorders have few objective findings, and patients often do not respond to well-established orthopaedic treatments, both nonsurgical and surgical. In some areas of the country the rate of reimbursement is low; that factor, combined with the excessive paperwork and the legal burden, discourages many orthopaedists from treating patients with these conditions. However, the incidence and cost of work-related disorders continue to increase, and the orthopaedic community is being called on to help understand their etiology and to attempt to control the "epidemic" that has significantly affected the survival of certain industries. The authors review the current orthopaedic and occupational medicine literature and suggest a management approach that has been found effective in reducing both costs and disability due to these disorders. PMID- 10795049 TI - Acute Pelvic Fractures: I. Causation and Classification. AB - Acute pelvic fractures are potentially lethal, even with modern techniques of poly-trauma care. The appropriate treatment of such fractures is dependent on a thorough understanding of the anatomic features of the pelvic region and the biomechanical basis of the various types of lesions. Although the anterior structures, the symphysis pubis and the pubic rami, contribute approximately 40% to the stiffness of the pelvis, clinical and biomechanical studies have shown that the posterior sacroiliac complex is more important to pelvic-ring stability. Therefore, the classification of pelvic fractures is based on the stability of the posterior lesion. In type A fractures, the pelvic ring is stable. The partially stable type B lesions, such as "open-book" and "bucket-handle" fractures, are caused by external- and internal-rotation forces, respectively. In type C injuries, there is complete disruption of the posterior sacroiliac complex. These unstable fractures are almost always caused by high-energy severe trauma associated with motor vehicle accidents, falls from a height, or crushing injuries. Type A and type B fractures make up 70% to 80% of all pelvic injuries. Because of the complexity of injuries that most often result in acute pelvic fractures, they should be considered in the context of polytrauma management, rather than in isolation. Any classification system must therefore be seen only as a general guide to treatment. The management of each patient requires careful, individualized decision making. PMID- 10795050 TI - Acute Pelvic Fractures: II. Principles of Management. AB - The past two decades have seen many advances in pelvic-trauma surgery. Provisional fixation of unstable pelvic-ring disruptions and open-book fractures with a pelvic clamp or an external frame with a supracondylar pin has proved markedly beneficial in the resuscitative phase of management. In the completely unstable pelvis, external clamps and frames can act only as provisional fixation and should be combined with skeletal traction. The traction pin is usually used only until a definitive form of stabilization can be applied to keep the pelvic ring in a reduced position. If the patient is too ill to allow operative intervention, the traction pin can remain in place with the external frame as definitive treatment. Symphyseal disruptions and medial ramus fractures should be plated at the time of laparotomy. Lateral ramus fractures can usually be controlled with external frames. A role has been suggested for percutaneous retrograde fixation of the superior pubic ramus; however, the benefits to be gained may not be enough to outweigh the serious risks of penetrating the hip, and this technique should therefore be used only by surgeons trained in its performance. The techniques for posterior fixation are becoming more standardized, but all still carry significant risks, especially to neurologic structures. PMID- 10795051 TI - Congenital Deformities of the Upper Extremity. AB - Congenital deformities of the upper extremity are rare. They are often associated with other, more severe disorders of the cardiovascular, craniofacial, neurologic, and musculoskeletal systems. Most upper-extremity congenital anomalies are minor and cause no functional deficits, and surgical reconstruction is therefore unnecessary. If a severe cosmetic deformity is present or there is significant functional compromise, surgical treatment is indicated. The authors review the common congenital deformities of the upper extremity and offer treatment recommendations. PMID- 10795052 TI - Stable Slipped Capital Femoral Epiphysis: Evaluation and Management. AB - Slipped capital femoral epiphysis (SCFE) has been classified traditionally on the basis of the duration of symptoms, but it has recently been recognized that this classification system may be misleading. It has instead been recommended that slips be classified on the basis of the presence or absence of gross instability between the epiphysis and the metaphysis. An adolescent with chronic SCFE has had symptoms for more than 3 weeks and does not have physeal instability. The first priority of treatment of stable chronic SCFE is to avoid the complications of avascular necrosis and chondrolysis while securing the epiphysis from further slippage. The treatment of choice for stable chronic SCFE is stabilization in situ, which can be most easily achieved with single-screw fixation. Primary realignment procedures, such as osteotomies, are not recommended. PMID- 10795053 TI - The Mangled Extremity: When Should It Be Amputated? AB - Amputation of a mangled extremity is repugnant to the patient and the surgeon. However, prolonged unsuccessful attempts at salvage are costly, highly morbid, and sometimes lethal. Much discussion has taken place regarding which criteria predict successful salvage, and predictive indices have been proposed in an attempt to identify limbs for which attempted salvage is unlikely to succeed. The Mangled Extremity Severity Score, or MESS, system is the most thoroughly validated of the various classification systems, but at present there is no predictive scale that can be used with confidence to determine whether to amputate or attempt to salvage a mangled lower extremity. Therefore, these systems should serve only as guides to supplement the surgeon's clinical judgment and experience. Although salvage for severe injuries below the knee can be difficult and the functional outcome unpredictable, prosthetic function after transtibial amputation is generally good. Conversely, prosthetic function after transfemoral or transradial amputation is often poor, while salvage of some useful function for injuries above the knee is often successful. When limb loss is inevitable, immediate amputation is desirable. If obvious criteria for primary amputation are not met, however, it is reasonable to consider an initial salvage attempt, observation, and subsequent early secondary amputation. PMID- 10795054 TI - Lateral Meniscal Variants: Evaluation and Treatment. AB - The normal lateral meniscus is morphologically more variable than the medial meniscus. The abnormal lateral meniscus also varies with respect to size, shape, and stability. Variations can occur in patients of all ages. The underlying causes of lateral meniscal abnormalities are multifactorial. The spectrum of abnormalities includes the most common variant, discoid lateral meniscus, as well as less common conditions, such as a lateral meniscal variant with absence of the posterior coronary ligament. Treatment should be based on the severity of symptoms and the type of pathologic lesion. PMID- 10795055 TI - Isthmic Spondylolisthesis in the Adult. AB - Isthmic spondylolisthesis is a common condition and is frequently identified in the adult patient with low back pain. Although the natural history of this condition is not well defined, it is a common indication for nonoperative and operative treatment. The authors outline a systematic approach to the evaluation of the adult patient with isthmic spondylolisthesis. If radiologic studies are required, magnetic resonance imaging has improved the visualization of nerve-root compression in the neural foramen and is now widely used. Nonoperative treatment is the preferred approach in most symptomatic patients and is successful in as many as 60%. If nonoperative treatment fails, surgery may be recommended. Arthrodesis continues to be the mainstay of surgical treatment. Nerve-root decompression can be used in selected patients with radiculopathy. Although the Food and Drug Administration still considers the use of pedicle-screw instrumentation investigational or experimental, it has gained wide acceptance as an adjunct to fusion in the adult. It is important to note, however, that such use has not yet been proved safe and effective. Reduction of the spondylolisthetic segment has increased in acceptance for a small subset of patients with defined indications but carries a significant risk of complications. PMID- 10795056 TI - Acute Compartment Syndrome: Update on Diagnosis and Treatment. AB - Acute compartment syndrome can have disastrous consequences. Because unusual pain may be the only symptom of an impending problem, a high index of suspicion, accurate evaluation, and prophylactic treatment will allow the physician to intervene in a timely manner and prevent irreversible damage. Muscles tolerate 4 hours of ischemia well, but by 6 hours the result is uncertain; after 8 hours, the damage is irreversible. Ischemic injury begins when tissue pressure is 10 to 20 mm Hg below diastolic pressure. Therefore, fasciotomy generally should be done when tissue pressure rises past 20 mm Hg below diastolic pressure. PMID- 10795057 TI - Hand Infections: Treatment Recommendations for Specific Types. AB - Hand infections can be associated with considerable morbidity. Expeditious treatment is needed to minimize permanent dysfunction, loss of work, and medical cost. Hand infections can affect the skin, subcutaneous tissues, fascia, subfascial and synovial spaces, joints, and bone. Pathogens include a variety of bacteria, viruses, yeasts, fungi, and mycoplasmata. Management frequently involves rest, elevation, incision and drainage, and appropriate antibiotic therapy. The orthopaedic surgeon must be knowledgeable about the pertinent anatomy and how this influences the behavior of specific types of infections, the role of immunocompromise, and the importance of early mobilization. PMID- 10795058 TI - Meniscal Injuries in Children and Adolescents: Diagnosis and Management. AB - Isolated meniscal injuries are uncommon in children under the age of 14, but the frequency increases thereafter. Meniscal tears in children are frequently associated with congenital meniscal abnormalities, while those in adolescents are often associated with ligamentous injuries of the knee. The combination of recurrent and often dramatic popping and intermittent episodes of locking has been termed the "snapping knee syndrome." This symptom complex is almost invariably associated with a discoid meniscus. Although double-contrast arthrography has proved to be a reliable diagnostic technique, magnetic resonance imaging is now the modality of choice. Treatment options for meniscal injuries in young patients should reflect a preference for repair over excision. The long term clinical results of total meniscectomy in children have demonstrated a high incidence of degenerative joint disease. Partial excision may provide better results. The efficacy of meniscus transplantation in the skeletally immature knee, although attractive, is as yet unproved and is therefore not an acceptable treatment option. PMID- 10795059 TI - Traumatic Anterior Glenohumeral Instability: The Role of Arthroscopy. AB - The static and dynamic stabilizers of the glenohumeral joint act together to ensure joint stability throughout the wide range of normal shoulder motion. These structures are functionally altered after a traumatic anterior shoulder dislocation, often resulting in recurrent instability. The arthroscope has enhanced the surgeon's ability to examine and repair the unstable glenohumeral joint. The rate of recurrence of instability after arthroscopic stabilization has historically been unacceptably high compared with that after open stabilization techniques. The authors offer a treatment algorithm and suggest guidelines for the use of arthroscopic techniques in the treatment of the unstable shoulder. PMID- 10795060 TI - Rheumatoid Arthritis of the Cervical Spine. AB - Cervical involvement in patients with rheumatoid arthritis occurs primarily in the upper cervical spine. The characteristic deformities are atlantoaxial subluxation, vertical settling, and subaxial subluxation. The typical patient complaints are neck pain and occipital pain. Subtle signs of myelopathy may also be present. Useful radiologic studies include plain radiography, tomography, and functional magnetic resonance imaging. The most helpful radiographic measurements are the anterior atlantodens interval, the posterior atlantodens interval, and assessment of vertical settling. Atlantoaxial subluxation greater than 9 mm with vertical settling and a posterior atlantodens interval less than 14 mm correlate with neurologic deficit. Nonoperative management does not change the natural history of cervical disease. Traditional surgical indications include intractable pain and neurologic deficit. The author discusses more controversial indications and proposes a rationale and protocol for treatment. The primary surgical objectives are to achieve stabilization of the affected segments and to relieve neural compression by reduction of subluxations or direct decompression. Arthrodesis provides reliable pain relief. Neurologic recovery occurs more consistently in patients with lower grades of preoperative myelopathy. PMID- 10795061 TI - Septic Arthritis of the Hip in Children. AB - Recent advances in the management of septic arthritis of the hip in children include a better understanding of the effects of infection on articular cartilage; improvements in diagnostic tests, including erythrocyte sedimentation rate, C-reactive protein analysis, and ultrasonography; and more efficacious home intravenous and oral antibiotic therapy. Early diagnosis is essential to successful treatment. Needle aspiration is the most specific diagnostic test; however, false-negative results are possible. Prompt surgical drainage and postoperative antibiotic therapy until signs of infection resolve are necessary to prevent late sequelae. Surgical treatment of limb-length inequality is more useful than attempts to salvage the destroyed or incongruent joint. PMID- 10795063 TI - Surgical Correction of Malunited Fractures of the Distal Radius. AB - Malunion of a distal radius fracture may result in functional limitation and pain. Reestablishing the geometry of the metaphysis and the alignment of the articular surface, restoring the relationship at the distal radioulnar joint, and respecting the soft-tissue envelope are elements of a successful reconstruction. The author describes a systematic approach to evaluation and treatment, including the indications for surgical correction, techniques of reconstruction, and potential complications. Salient aspects of postsurgical rehabilitation and salvage procedures are discussed. PMID- 10795062 TI - Nerve Entrapment Syndromes of the Foot and Ankle. AB - Nerve entrapment at the ankle and midfoot is an important, yet frequently underrecognized, source of foot pain. Familiarity with the nerve anatomy and known entrapment sites facilitates evaluation; local nerve lesions in the foot and ankle must be distinguished from proximal nerve dysfunction and systemic diseases that may affect nerve function. Treatment is directed toward reducing contact pressure from footwear and addressing other underlying problems, such as edema and ankle instability that may be contributing to nerve dysfunction. PMID- 10795064 TI - Hip Osteotomies: A Biomechanical Consideration. AB - Proximal femoral and periacetabular osteotomies for the treatment of osteoarthrosis have produced varying clinical results. Because the underlying pathomechanics of osteoarthrosis are not yet fully understood, it is difficult to predict which type of osteotomy will most improve the biomechanical environment in a given situation. Osteotomies result in relatively small changes in joint load (10% to 20% at most), which probably do not govern tissue responses. However, osteotomies do change the distribution of the load (i.e., stresses) and, perhaps more important, stress gradients. These changes in contact and underlying stresses undoubtedly affect cartilage and bone adaptation. It is likely that the magnitude of stress and strain in cartilage and underlying bone can be altered in such a way that more predictable beneficial remodeling will occur. Emerging imaging and computational technologies may allow patient-specific modeling, which should improve the efficacy and durability of repair. PMID- 10795065 TI - Fatigue Fractures of the Femoral Neck in Athletes. AB - Fatigue fractures of the femoral neck are potentially disabling injuries if the diagnosis is missed or delayed and proper treatment is not provided. Previously considered primarily an injury of military personnel, femoral-neck fatigue fractures are becoming increasingly more common among nonmilitary athletes. The pathogenesis of this condition is multifactorial. Ultimately, fracture is thought to be caused by repetitive submaximal stresses that occur with a frequency that exceeds the adaptive ability of the bone. The clinical hallmark of a femoral-neck fatigue fracture in an impact-loading athlete is nonspecific, activity-related hip pain that is relieved by rest. Plain radiographs obtained initially are often negative. Radionuclide imaging is useful but not specific. Magnetic resonance imaging has demonstrated better specificity, sensitivity, and accuracy than are available with other modalities in the diagnosis of this injury. Treatment of compression-side injuries is generally conservative. Treatment of tension-side injuries remains controversial. The authors recommend aggressive treatment of tension-side injuries to prevent the potentially catastrophic sequelae of displacement, which include osteonecrosis, malunion, and coxarthrosis. PMID- 10795066 TI - Reflex Sympathetic Dystrophy in the Upper Extremity. AB - The diagnosis and treatment of pain are among the most challenging problems facing orthopaedic surgeons, and reflex sympathetic dystrophy is probably the most frustrating and difficult pain syndrome to manage. Pain, swelling, and autonomic dysfunction are cardinal signs of the condition. Although the pathogenesis is still unclear, many theories have been proposed. Because reflex sympathetic dystrophy is sympathetically mediated, diagnosis can be confirmed on the basis of response of the pain to sympathetic blockade. Treatment may include an appropriate exercise program, a-adrenergic blocking agents, mood-elevating drugs, calcium channel blockers, intravenous regional blocks, and stellate ganglion blocks. Recent additions to therapy include electroacupuncture, transcutaneous electrical nerve stimulation, and biofeedback. Prognosis is, at best, guarded with this perplexing condition, but the best response is obtained when diagnosis is made early (within the first 2 or 3 weeks after injury) and treatment is initiated during the first stage of the disease. PMID- 10795067 TI - Patellar Fractures: Contemporary Approach to Treatment. AB - Patellar fractures are a diverse group of injuries with a variety of fracture patterns. The surgical goals are anatomic reconstruction of the articular surface and stable fixation to allow early motion. Contemporary methods of treatment include screws, the modified tension band, and a combination of the two. Screws and wires have demonstrated specific failure patterns in experimental models, and biomechanical studies suggest that the combination of screws and wires provides more secure fixation. Partial patellectomy is the procedure of choice in comminuted fractures that cannot be anatomically reduced. Optimal treatment of patellar fractures requires individualization of surgical technique. The postoperative rehabilitation program should be based on the intraoperative findings and the stability of fixation. PMID- 10795068 TI - Rheumatoid Arthritis of the Hip. AB - The hip joint may be affected in 15% to 28% of all patients with rheumatoid arthritis. Radiographic evidence of involvement includes periarticular osteopenia, cystic changes, and a variable amount of progressive protrusio acetabuli. Histomorphometric study has shown increased bone turnover in acetabular biopsy specimens from rheumatoid patients undergoing total hip arthroplasty. Due to the relative fragility of bone in these patients, there is an increased risk of fracture of the proximal femur due to minor trauma, and a high rate of loss of fixation has been reported. Total hip arthroplasty has been successful in the treatment of severe rheumatoid arthritis of the hip in patients of all ages. Special attention should be paid to the cervical spine and the patient's medical treatment regimen during the preoperative evaluation. Cemented total hip arthroplasty has been associated with a higher prevalence of late infection and acetabular component loosening in rheumatoid patients than in osteoarthritic patients. Loosening of cemented components is accelerated in patients with juvenile rheumatoid arthritis. Several short-term studies have documented successful early results with noncemented components in patients with rheumatoid arthritis; however, longer-term studies are necessary to determine whether the improvements in function and survival are greater than with cemented components. PMID- 10795069 TI - Osteonecrosis of the Humeral Head. AB - Current understanding of osteonecrosis of the humeral head is largely based on previous studies of the femoral head. Similarities between the two sites are numerous, but the shoulder has many unique characteristics. The anatomy of the glenohumeral joint, motion at the scapulothoracic joint, the rich vascular supply of the surrounding soft tissues, and the accommodations for the different forces (shear, compression) exerted across the glenohumeral joint all allow the shoulder to tolerate a greater amount of deformity. A number of pathologic agents can cause bone death by disrupting the blood supply, among them corticosteroid use, trauma, dysbarism, hemoglobinopathies, and various systemic diseases that disrupt the vascular system, such as Gaucher's disease and systemic lupus erythematosus. Management is similar to that of femoral osteonecrosis; the earlier stages respond well to nonoperative approaches, and the more advanced stages require surgical intervention. Hemiarthroplasty and total shoulder arthroplasty have produced good outcomes. Surgical intervention with core decompression, vascular flaps, and arthroscopic debridement have also shown promise, but further studies are necessary to define their optimal use. PMID- 10795070 TI - Oral levofloxacin in the treatment of community-acquired pneumonia. AB - We evaluated the efficacy of oral levofloxacin (500mg/day for 7-21 days) in the treatment of 20 adults patients with community-acquired pneumonia (CAP) requiring hospitalization in an open prospective study. The microbiological cause of the pneumonia was identified in 14/20 patients using lower respiratory tract secretions obtained by bronchoscopy (12) and/or blood culture (2). Eight patients had S. pneumoniae, 2 P.aeruginosa, 1 H.influenzae, 1 S.aureus, 1 mixed S. aureus and K.pneumoniae, and 1 E.coli and Grp.D Streptococcus. All of the patients evaluated were judged to be improved or cured. Levofloxacin is an additional option as monotherapy for the treatment of CAP. PMID- 10795071 TI - Infectious and neoplastic disorders of the thyroid in AIDS patients: an autopsy study. AB - The course of human immunodeficiency virus infection and the acquired immunodeficiency syndrome can be complicated by a variety of endocrine abnormalities. This article describes the findings of a prospective autopsy study of the thyroid in 100 patients who died of complications of AIDS before the advent of the so-called highly active antiretroviral therapy [corrected]. A wide range of bacterial, fungal, viral, and neoplastic disorders were observed. Mycobacterium tuberculosis was recorded in 23% of the patients, cytomegalovirus in 17%, Cryptococcus in 5%, Mycobacterium avium in 5%, Pneumocystis in 4%, and other bacteria or fungi in 7%. Kaposi's sarcoma was recorded in 2% of patients and occult papillary carcinoma in 4%. Four patients had dual infections of the thyroid. The mean weight of the thyroid was lower than normal, and 1 case of thyroid follicular atrophy is presented. A review of the medical literature on thyroid disorders in HIV-infected patients is included. Physicians caring for HIV patients should be aware of the possibility of thyroid dysfunction in their patients. PMID- 10795072 TI - Assessing the antimicrobial prescription request process in a teaching hospital in Brazil: regulations and training. AB - It is known that antimicrobials are often prescribed inappropriately. One method used to deal with this problem is to regulate antimicrobial use by monitoring prescriptions. We report a study of physician compliance with the request for antibiotic process which was prepared and reviewed by a special infection control committee in our hospital the Nosocomial Infection Prevention Service (SCIH). The objective of this study was to identify the profile of inappropriate requests for restricted therapeutic antimicrobials used at Nossa Senhora da Conceicao Hospital (HNSC), in Porto Alegre, Brazil. All 3,389 requests for therapeutic antimicrobials made between May 20, and October 31, 1996, were assessed and classified as appropriate and inappropriate. We determined that 17. 8% of the requests were inappropriate (a total of 720 errors). These were categorized according to 12 reasons for inappropriateness. Of these, the 3 most frequent inappropriate requests were deviation from standard use (26.73%), inappropriate length of treatment (23. 19%), and unfounded justification (13.61%). The reasons for inappropriateness were also arranged in three categories considering the following aspects: I. technical (59%); II. compatibility with the institutional program (32.36%), and III. administrative (8.61%). The 720 requests that were initially rejected were evaluated to see how antibiotic use was affected. In approximately 400 (55%), the forms could be appropriately modified after discussion with the physician. We conclude from this study that most of the inappropriate requests for antimicrobials in our hospital can be remedied by educating the staff since the errors were largely technical in nature. Thus, the SCIH should focus more on its role as an educational rather than as a regulatory body. By expanding this educational role, we anticipate improved physician compliance with our guidelines, and more appropriate antimicrobial prescribing and usage. PMID- 10795073 TI - A retrospective bacteriological study of mycobacterial infections in patients with acquired immune deficiency syndrome (AIDS). AB - The main species of mycobacteria isolated in 51 patients with acquired immune deficiency syndrome (AIDS) admitted to the Clinical Hospital of UNICAMP (Teaching Hospital) in 1996, were studied retrospectively by recording the isolation site and signs of pathogenesis. Of these isolates, 31 (55%) were M.tuberculosis and 20 (45%) M.avium complex. Strains of M.tuberculosis isolated in 1996, 1997, and 1998, originating from 42 patients who had AIDS, were tested for susceptibility to isoniazid and rifampicin. Of these, 34 (81%) were susceptible to both drugs tested, 4 (10%) were resistant to isoniazid and susceptible to rifampicin, 2 (5%) were susceptible to isoniazid and resistant to rifampicin, and 2 (5%) were resistant to both rifampicin and isoniazid. We conclude that, in Brazil, M. avium complex infections in AIDS are more common than has been previously suggested; i.e., almost as frequent as M.tuberculosis infections. Approximately 20% of M.tuberculosis strains showed resistance to rifampicin and/or isoniazid. Further distribution of information regarding how to treat the disease in AIDS patients is needed. PMID- 10795074 TI - Antimicrobial resistance in Brazil: comparison of results from two multicenter studies. AB - To evaluate whether our previous study of the antimicrobial resistance patterns in three centers in Brazil represented the pattern in the country as a whole, the results were compared to new data on 855 isolates from 20 clinical laboratories and 36 hospitals located in different regions of Brazil. Both multicenter studies showed high rates of antimicrobial resistance among Gram-negative bacilli isolated in Brazilian hospitals with the most important problems being: 1) E.coli and K.pneumoniae that produce ESBL; 2) Enterobacter spp. which likely express chromosomally mediated (AmpC) stably derepressed cephalosporinases producing resistance to third-generation cephalosporins and broad spectrum penicillins; 3) carbapenem resistance among Acinetobacter spp. and P.aeruginosa; and 4) fluoroquinolone and aminoglycoside resistance among many Gram-negative species. Our results emphasize the importance of regional antimicrobial resistance surveillance programs in guiding empirical therapy and for focusing intervention controls of antimicrobial resistance. Although the SENTRY Program has only three participating centers in Brazil, its results were validated by a larger Brazilian multicenter study. PMID- 10795075 TI - Primary anetoderma. AB - The authors report a case of primary anetoderma in a 27-year-old black woman. Primary anetoderma is characterized by circumscribed oval plaques with overlying loose, wrinkled skin appearing as a depression or pouch-like protrusion of the skin. PMID- 10795076 TI - Calcinosis cutis following the administration of intravenous calcium therapy. AB - Calcinosis cutis, the cutaneous deposition of calcium salts in the dermis, can occur through a variety of pathogenetic mechanisms, and can be associated with both normal and elevated calcium levels. Iatrogenic causes of calcinosis cutis include extravasation of intravenously administered calcium chloride or calcium gluconate, and traumatic deposition of calcium in the skin, subsequent to electromyography or electroencephalography. We report two cases of calcinosis cutis following intravenous infusion of a calcium-containing salt. PMID- 10795077 TI - Managed care. PMID- 10795078 TI - Lipedematous alopecia. AB - Lipedematous alopecia is an unusual disorder characterized by short, brittle scalp hairs associated with thickening of the scalp by adipose tissue. All except one reported case have been in African-American females. It is our purpose to report the seventh case of this condition. PMID- 10795079 TI - Generalized annular borderline tuberculoid leprosy and update in management of Hansen's disease. AB - We describe a patient with widespread borderline tuberculoid leprosy and significant peripheral nerve involvement. Despite the presence of widespread lesions, Fite stains and polymerase chain reaction studies were initially negative. We discuss the diagnosis and treatment of leprosy including recent changes in treatment regimens and duration. PMID- 10795080 TI - Complete remission of refractory dyshidrotic eczema with the use of radiation therapy. AB - Dyshidrotic eczema is a chronic, enigmatic condition that usually affects the hands and feet. Modern-technique radiation therapy using megavoltage equipment for the treatment of dyshidrotic eczema has not been described in the literature before. A dramatic clinical response to low-dose external beam radiation therapy was observed in a patient refractory to multiple forms of topical and systemic agents. Complete resolution of this severe presentation of dyshidrosis occurred within 1 month following therapy, with a durable response at 6 months. Withdrawal of oral steroids, without flare of disease, was possible after 6 weeks, with the patient remaining free of medication at the 6-month interval. Complete remission of severe dyshidrotic eczema is achievable using low-dose external beam megavoltage therapy in situations where other forms of conventional therapies have failed. Lasting remission may allow for the complete withdrawal of oral or topical agents, which may become harmful with chronic use. PMID- 10795081 TI - Carcinoma erysipeloides from prostate cancer presenting as cellulitis. AB - We report a 72-year-old man with carcinoma erysipeloides presenting as cellulitis of the thigh. Immunohistochemical study of the skin biopsy showed positive staining to prostate-specific antigen, confirming that the tumor originated in the prostate. PMID- 10795082 TI - Verrucous carcinoma occurring in a lesion of oral lichen planus. AB - Verrucous carcinoma of the oral cavity is relatively rare. Well-documented associations include human papillomavirus and carcinogens such as tobacco. Less well understood is the association with chronic inflammation, such as seen in lichen planus. Verrucous carcinoma has previously been described occurring in lesions of lichen planus of the foot and penis. We report the first case, to our knowledge, of verrucous carcinoma occurring in a lesion of oral lichen planus. PMID- 10795083 TI - Erythrocyanotic discoloration of the toes. AB - A 35-year-old woman from Virginia presented with purplish, tender toes. She was diagnosed with pernio and successfully treated, initially with nifedipine and subsequently with management of the affected area's environment. Clinical presentations and treatment strategies are discussed below. PMID- 10795084 TI - The smell of burnt toast: a case report. AB - A 71-year-old woman awoke one morning to find that she perceived all aromas, odors, and fragrances as smelling like burnt toast. Over the next three years, numerous studies and therapeutic trials failed to elicit the cause of her dysosmia or to provide relief. Finally, the demonstration of small infarcts as seen on a brain MRI suggested that an infarct near the olfactory pathway was responsible for the sudden onset and the 11 year persistence of her unique paromia. PMID- 10795085 TI - Lamivudine (3TC)-induced contact dermatitis. AB - The nucleoside analogue lamivudine (3TC) is commonly used in multidrug therapy of human immunodeficiency virus-1 disease because it not only potentiates the antiviral effects of other reverse transcriptase inhibitors, but it is also relatively nontoxic. We present a patient who developed a contact dermatitis to lamivudine after prolonged exposure. PMID- 10795086 TI - An epidemiologic and therapeutic reassessment of scabies. AB - Scabies is a highly contagious infestation that causes considerable discomfort. Newer information concerning therapeutics and epidemiologic dogma needs assessment. An epidemiologic evaluation of available world data is analyzed, as well as an assessment of therapeutic alternatives. Using epidemiologic techniques, the following points are proven: scabies is not primarily a sexually transmitted disease; 30-year cycles of scabies do not exist; scabies spreads in households and neighborhoods in which there is a high frequency of intimate personal contact or sharing of inanimate objects; and fomite transmission is a major factor in household and nosocomial passage of scabies. Epidemiologic evaluation proves the necessity of fomite precautions and of treatment of asymptomatic family members and physical contacts of all cases of scabies. Single oral-dose therapy of ivermectin (Stromectol) appears to be the treatment panacea for this infestation. PMID- 10795087 TI - Acral lipoblastoma. AB - Lipoblastomas are rare, benign childhood tumors. We report a case of lipoblastoma on the heel of a 5-year-old girl who presented with a progressively enlarging, xanthoerythematous plaque. Surgical excision with follow-up for possible recurrence is the treatment of choice. PMID- 10795088 TI - Orthogonal acceleration time-of-flight mass spectrometry. AB - The principles and applications of time-of-flight mass spectrometry involving instruments with independent (orthogonal) axes for ion generation and mass analysis are reviewed. This approach, generally referred to as orthogonal acceleration time-of-flight mass spectrometry, has proved particularly advantageous for the combination of continuous ionization sources with time-of flight mass spectrometry. The history of the technique is briefly discussed along with the instrumental principles pertaining to all the stages of the instrumentation from ion source to detector. The applications of commercial and customized instruments are discussed for several ionization methods including electrospray, matrix assisted laser desorption/ionization, electron ionization, and plasma ionization. PMID- 10795089 TI - [Stroke. Options for primary prevention]. AB - Age, gender, race, and genetic factors count among the nonmodifiable risk factors for stroke. But primary prevention is possible through modification of other vascular risk factors. The treatment of arterial hypertension, with optimal values around 135/85 mmHg, reduces the risk of stroke by 50%. A significant risk reduction for ischemic stroke is possible with at least 30 minutes of physical activity twice a week, cessation of cigarette smoking, and treatment of hypercholesterolemia with statins. Dietary measures should include a reduction of animal proteins, normalization of body weight and a large amount of fruit and vegetables; small amounts of wine are allowed. A hyperhomocystinemia is treated with folic acid. Low dose estrogens (< 50 micrograms) do not increase the risk of ischemic stroke in young females, but are capable of reducing hypercholesterolemia in postmenopausal women if triglycerides are normal. Neither primary prevention of stroke with antiagreggants nor surgery for asymptomatic carotid artery stenoses is recommended as preventive treatment, but low dose anticoagulation is the prophylaxis of choice for atrial fibrillation. PMID- 10795090 TI - [Cortical dysgenesis. Current views on pathogenesis and pathophysiology]. AB - Cortical dysgenesia are a heterogenous group of genetic or acquired disturbances of cortical development which, due to the enormous progress in modern neuroimaging techniques, are increasingly recognized in association with a variety of clinical disorders. The spectrum of clinical manifestations, depending on type and extent of the alterations, includes severe mental retardation and epilepsy as well as neuropsychological deficits and some psychiatric disorders. Although pathogenesis and pathophysiology of cortical dysgenesis are still not fully understood, the recent discovery of responsible genes, growth factors, neurotransmitters, and exogenous factors sheds light on elementary mechanisms. The development of animal models mimicking different types of human cortical malformations helped to increase further the understanding of functional consequences of focal cortical dysgenesis. Several studies on these models reveal widespread alterations of cortical connectivity and excitability which are probably of crucial importance in different clinical disorders. PMID- 10795091 TI - [Neurological complications after organ transplantation]. AB - Following organ transplantation, 30-60% of patients develop neurologic complications which can be classified as pre-existing deficits due to the underlying disease, complications during surgery, metabolic encephalopathies, neurotoxicity of immunosuppressant agents, opportunistic CNS infections, and secondary malignomas as indirect side effects of immunosuppression. While encephalopathies, seizures, or CNS infection can occur in all types of transplantation, some specific neurological complications exist for different types of organ transplantation. In this review, the clinical symptoms and treatment of both the common neurological complications as well as the particular neurological syndromes after liver, heart, and bone marrow transplantation are discussed. PMID- 10795092 TI - [Early rehabilitation after cerebral lesions: indications, methods, results]. AB - Early neurological and neurosurgical rehabilitation after severe brain injury of traumatic, hemorrhagic, ischemic, neoplastic, inflammatory, or hypoxic origin fills the gap between acute treatment and traditional rehabilitation services. During the last years, specific treatment modalities began to develop, directed at the key problems of severely brain injured patients: for example, stabilization of vegetative regulation, treatment of spasticity, swallowing disorders, disturbances of perception and communication, coping, and conditioning of the relatives. Procedures are gradually emerging to facilitate assessment, documentation of progress, and results. Prognosis is still very uncertain, especially in patients in a vegetative state. As a rule of thumb, about three out of four patients will improve substantially during the treatment. PMID- 10795093 TI - [Microsurgery of lumbar disc prolapse. Superior results of microsurgery as compared to standard- and percutaneous procedures (review of literature)]. AB - In a meta-analysis of 69 prospective and retrospective studies, we investigated the value of various surgical techniques in the treatment of lumbar disk herniations. This analysis includes standard diskectomy (5080 patients from nine series), microdiskectomy (5354/23), and comparison of both techniques (2494/10) and furthermore chemonucleolysis (2729/16), laser therapy (881/3), percutaneous nucleotomy (3506/18), comparisons of percutaneous techniques (942/5) with microdiskectomies (561/5) and standard diskectomies (1020/6). Outcomes were rated according to Macnab's criteria: I "excellent", II "good", III "improved", IV "same as before", V "worse", I/II "markedly improved", III "partially improved", IV/V "not improved", I-III "successful", and IV/V "unsuccessful". Results after microdiskectomy were "successful" more often (90% vs. 95%), "good/excellent" more often (82% vs. 73%), and patients left the hospital sooner than with standard diskectomy. Recurrence rates were comparable (4%). Results of both open techniques were clearly superior to those from any type of percutaneous treatment: chemonucleolysis und laser therapy were "successful" in 69% and 70% of cases respectively, with recurrence rates of 17% and 18%, endoscopic nucleotomy was "successful" in 84% of cases, and recurrence rate was 14%. Please ask the author for the reference database. PMID- 10795094 TI - [Coping behaviors in Parkinson's disease]. AB - In 45 patients with Parkinson's disease, we investigated coping behavior and its correlations to demographic and disease-related data, locus of control, depression, and psychosocial adaptation. Active, problem oriented, and self reorganizing strategies were predominantly used and regarded as especially helpful by the 27 men and 18 women (age: 56 years; duration of illness: 9 years). While age and sex were not associated with coping, external locus of control correlated positively to "depressive coping" and duration of illness correlated negatively to "distraction and self-affirmation." Coping strategies regarded as maladaptive and a small degree of internal locus of control correlated to unfavourable results for depression and satisfaction. PMID- 10795095 TI - [MRI spectroscopy, T2 relaxometry, and postoperative prognosis in cryptogenic temporal lobe epilepsy]. AB - The preoperative localization of pharmaco-resistant focal epilepsies before surgery and the prognosis concerning seizure outcome are both of importance. In addition to conventional MRI diagnostics for the detection of small epileptogenic lesions, proton magnetic resonance spectroscopic imaging (HMR spectroscopy) can be useful for assessing the bilaterality of pathological changes in cryptogenic temporal lobe epilepsies. The technique and findings of HMR spectroscopy are reported in patients with cryptogenic temporal lobe epilepsies. The findings indicate that chemical shift imaging (CSI) investigations of the ipsilateral and contralateral hemispheres provide important information for the prediction of seizure outcome after epilepsy surgery. PMID- 10795096 TI - [Projection of the number of multiple sclerosis patients in Germany]. AB - For determining the prevalence of multiple sclerosis (MS) in Germany, the outcomes of several epidemiological studies are available. If these rates are transferred to Germany as a whole, figures result of 67,000 to 138,000 MS patients in all. The differing prevalence rates may be caused by various problems in finding patients, the epidemiological area, and different population structures. To avoid these difficulties, a new approach for calculating the number of patients with diagnosed MS was chosen based on representative samples taken from the groups of physicians involved in MS treatment. The disadvantage of this approach is that it does not allow any questioning of the diagnosis. Projecting the results in this way and their subsequent compilation--taking "double treatments" into consideration--showed a total of 122,000 patients with diagnosed multiple sclerosis in Germany. This figure corresponds to a prevalence rate of 149.1 per 100,000 inhabitants and is within the range found by Poser et al. [19]: 127 per 100,000 excluding the diagnostic category "possible multiple sclerosis" and 170 per 100,000 including it. PMID- 10795097 TI - [Relapsing paraneoplastic cerebellar syndrome and limbic encephalitis in a patient with breast cancer]. AB - Paraneoplastic neurological syndromes are rare diseases that occur in relation to cancer. Supporting the hypothesis of an autoimmune mechanism, specific antineuronal antibodies have sometimes been detected. The current possibilities for treatment are limited. A female patient, aged 57, suffering from a breast cancer, developed a severe paraneoplastic cerebellar syndrome and limbic encephalitis within a few weeks. It is possible that the impressive partial remission that occurred during the ensuing 6 months was not due to therapy. Although the patient was still bound to a wheelchair, discharge from hospital was possible because she was still able to perform daily tasks by herself. A recurrence of the cerebellar symptoms with mild alterations of mental status occurred 2 months later but again showed a good remission. PMID- 10795098 TI - [Cerebral vasculitis as a concomitant neurological illness in Crohn's disease]. AB - In Crohn's disease, some concomitant neurological illnesses such as cerebral ischemia following arterial or venous thrombosis, subacute combined degeneration of the spinal cord following malabsorption of vitamin B12 or folic acid, opticus neuropathy, and polyneuropathy have been described. Cerebral vasculitis secondary to Crohn's disease seems to be a very rare phenomenon. We report on three such cases in three female patients (aged 26, 29, and 61 years). All patients became symptomatic with a hemiparesis; one complained additionally of a speech disorder, headache, and intermittent loss of orientation. In CT and MRI scans, multiple lesions were detected; cerebral angiography showed multiple stenoses of middle- and large-sized vessels that were compatible with cerebral vasculitis. Serologic tests concerning vasculitis were inconspicuous at that time. Under anticoagulation (in two cases) and immunosuppressive therapy, neurologic symptoms disappeared. In the following 6 to 12 months, no new neurological symptoms appeared. In two cases, Doppler sonographic controls showed stationary and, in one case, progressive intracranial stenoses. Since autoimmunologically caused inflammatory bowel diseases might be associated with vasculitis of other organs, the appearance of cerebral vasculitis secondary to Crohn's disease is a possible organ manifestation by inflamed vessels. PMID- 10795099 TI - ["Ghost and mimicry-tumor"--primary CNS lymphoma]. AB - In recent years, the frequency of primary cerebral lymphoma (PCNSL) has increased, even among immunocompetent patients. In order to treat the disease optimally, early diagnosis is important. We present three patients with atypical courses of this disease and stress the importance of PCNSL in the differential diagnosis for optimal treatment. In a 75-year-old man, a space-occupying, radiopaque, enhancing CNS lesion disappeared completely after biopsy and short steroid therapy. One year later, the tumor recurred on the other side and again regressed after steroid therapy. The first biopsy showed signs of a papillary tumor, so a choroid plexus papilloma was suspected initially. A 57-year-old woman developed progressive bilateral hearing dysfunction. Lymphocytic pleocytosis led to a primary diagnosis of chronic lymphocytic meningitis. During the further course of disease, the patient developed multiple space-occupying cerebral lesions. Stereotactic biopsy revealed PCNSL. Despite combined chemo- and radiotherapy, a relapse occurred. A 49-year-old woman rapidly developed memory and concentration disturbances. Computed tomography revealed diffuse edema in both hemispheres and MRI detected severe, diffuse, white matter lesions. The CSF revealed lymphocytic pleocytosis including plasma cells. Herpes encephalitis and, after lack of clinical improvement and progression of the MRI changes, acute disseminated encephalomyelitis (ADEM) were suspected. Corticosteroid treatment was initiated. After enormous clinical improvement, a clinical relapse occurred and MRI detected bitemporal and singular space-occupying lesions in the corpus callosum and hypothalamus. Finally, open biopsy showed PCNSL. However, on examination of the treatment history of patient 1, the initial diagnosis must be revised; a PCNSL seems most probable. The phenomenon of tumor remission under steroid administration is rare. In patients 2 and 3, atypical clinical signs and symptoms delayed diagnosis of PCNSL. This tumor can mimic diverse neurological diseases and remit following corticosteroid treatment alone. In unclear cerebral disease, biopsy should be performed early for exact diagnosis and optimal treatment. PMID- 10795100 TI - [Speech disturbances and dyskinesias as initial symptoms of cortico- basal degeneration]. AB - The article describes the development of symptoms in a 59-year-old patient. Dyskinesia and speech disorder were the only clinical features in the beginning. Increased immunological parameters and only slight hypokinetic-rigid signs for a long time made the diagnostical and therapeutical process more difficult, as well as atypical findings in neuroimaging techniques. Corticobasal degeneration was diagnosed about 6 years after onset of clinical symptoms. PMID- 10795101 TI - [Isolated reversible unilateral paresis of hypoglossal nerve in tonsillitis--case report]. AB - We report on a 30-year-old patient with isolated, left-sided hypoglossal nerve palsy after uncomplicated, presumably streptococci-induced tonsillitis. Needle electromyography (EMG) of the tongue showed denervation changes in the muscles supplied by the left hypoglossal nerve. Cranial CT and MRI, CSF examination, Doppler sonography, visual evoked potential (VEP), and auditory evoked potential (AEP) showed no abnormalities and, in particular, no signs of the carotid artery dissection or brainstem lesion. The symptoms and signs resolved within a few weeks after penicillin V treatment. We suppose that the hypoglossal affection of the uncomplicated tonsillitis in this case was due to an aberrant position of the nerve. PMID- 10795102 TI - [Comments on the contribution by M. Krause et al. Subsequent therapeutic management after stroke--an assessment of current status]. PMID- 10795103 TI - [Concept of interregional and regional stroke treatment in Germany. Report of the recommendation of Commission 1.06 "Stroke Units and Acute Stroke Therapy" of the German Society for Neurology]. PMID- 10795104 TI - [Hans Dieter Lux. Comments on cover portrait]. PMID- 10795105 TI - The role of the Centers for Disease Control and Prevention in dialysis. PMID- 10795106 TI - Role of the CDC in hemodialysis: an historical perspective. PMID- 10795108 TI - An evolving partnership: the role of the Centers for Disease Control in the dialysis community. PMID- 10795107 TI - Centers for Disease Control and Prevention: a clinician's view of the past, present, and future. PMID- 10795109 TI - National surveillance of dialysis-associated diseases in the United States, 1997. PMID- 10795110 TI - CDC investigations of noninfectious outbreaks of adverse events in hemodialysis facilities, 1979-1999. PMID- 10795111 TI - Outbreaks of infection and/or pyrogenic reactions in dialysis patients. AB - These dialysis-related outbreaks demonstrate the ongoing potential for infection related morbidity and mortality among dialysis patients. Many of these outbreaks could have been prevented by adequate water treatment, proper disinfection of water systems and dialysis machines, adherence to recommended reprocessing protocols in centers reusing dialyzers, and more stringent quality control monitoring. Finally, these outbreaks highlight the importance of active surveillance for adverse events among dialysis patients. The incidence of gram negative bacteremia, pyrogenic reactions, and peritonitis should be monitored over time and any increase in incidence investigated. PMID- 10795112 TI - Description of a new surveillance system for bloodstream and vascular access infections in outpatient hemodialysis centers. AB - Bloodstream and vascular access infections are a threat to hemodialysis patients. However, there are few studies of rates of such infections and there are no standardized methods for ongoing data collection. Because of frequent hospitalizations and receipt of antimicrobials, hemodialysis patients are at high risk for infection with drug-resistant bacteria. This article describes a new voluntary national surveillance system. Each month participating dialysis center personnel will record the number of chronic hemodialysis patients that they treat (broken down into four types of vascular access). A one-page form will be completed for each hospitalization or in-unit IV antimicrobial start among these patients. These data will allow calculation, stratified by type of vascular access, of several rates, including hospitalizations, in-unit IV antimicrobial starts, and vascular access infections. For individual dialysis centers, this surveillance system will provide a simple and standardized method for recording data, calculating rates, and comparing rates over time. It is hoped that collection and examination of these data will lead to quality improvement measures. For government and the medical and public health communities, aggregation of these data from many dialysis centers will provide a wealth of information that is not currently available. For further information, or to receive a protocol for this study, contact Elaine R. Miller, RN, MPH, at (404)639 6422 (telephone), (404)639-6459 or 6458 (fax), or erm4@cdc.gov (e-mail:). Information is also available on the CDC website at http:@www.cdc.gov/ncidod/hip/Dialysis/dialysis.+ ++htm. PMID- 10795113 TI - Vaccine recommendations for patients on chronic dialysis. The Advisory Committee on Immunization Practices and the American Academy of Pediatrics. AB - Pediatric patients on dialysis should receive all the vaccines currently recommended by the ACIP and the AAP for healthy children, except the oral polio vaccine (34, 35). Adult patients should receive the hepatitis B vaccine series, pneumococcal vaccine, yearly influenza vaccinations, tetanus-diphtheria toxoids, and varicella vaccine, if they are susceptible (33, 48, 69). Vaccines are well tolerated by these patients (33), but higher doses and/or additional boosters may be required periodically to adequately protect dialysis patients from vaccine preventable diseases (33, 36, 37, 82, 83). Following vaccination, antibody concentrations for hepatitis B vaccine should be measured annually and booster doses administered when antibody concentrations fall below protective levels (33, 38). Although both children and adults on dialysis may show an impaired and/or delayed immunologic response to certain antigens, particularly hepatitis B virus and S. pneumoniae, appropriate immunizations can significantly reduce the risk of serious complications from vaccine-preventable diseases (11, 84). Because the protection these vaccines provide may be incomplete or transient, infection control strategies at hospitals and other health care facilities should be implemented simultaneously. Health care providers are encouraged to assess each patients need for vaccinations individually and formulate immunization strategies early in the course of progressive renal disease, ideally before the patient requires dialysis. PMID- 10795114 TI - Hepatitis G virus infection among hemodialysis patients: true infection or innocent bystander? PMID- 10795115 TI - Hematocrit level associated mortality in hemodialysis patients, by Ma JZ, Ebben J, Xia H, Collins AJ. J Am Soc Nephrol 10:610-619, 1999. PMID- 10795116 TI - [A double-blind comparative study of the effectiveness and tolerance of paroxetine and amitriptyline in treatment of breast cancer patients with clinically assessed depression]. AB - OBJECTIVE: In this double-blind, non-placebo controlled study [corrected], 179 patients with treated breast cancer who fulfilled the ICD-10 criteria for an acute depressive episode underwent an 8-week course of antidepressant treatment with either the tricyclic amitriptyline (75-150 mg, n = 87) or the serotonin reuptake inhibitor paroxetine (20-40 mg, n = 88). METHODS: The change in clinical status relative to baseline was measured with the Montgomery-Asberg Depression Rating Scale (MADRS), the Clinical Global Impression (CGI), the Functional Living Index-Cancer (FLIC) and the Patient Global Evaluation. RESULTS: Both treatment groups showed significant improvement in all parameters at weeks 3, 5 and 8. At no time was there a significant difference in the efficacy of the antidepressants used. Adverse events, most of which were transitory, were reported by 53% of the patients in the paroxetine group and 60% in the amitriptyline group. The 8-week treatment was completed by 81% of the paroxetine and 76% of the amitriptyline patients. CONCLUSIONS: The results of this study show that depression in breast cancer patients can be correctly diagnosed and adequately treated by non psychiatrists. The treatment with both medications was carried out in dose ranges which correspond to that employed in physically well patients. PMID- 10795117 TI - [Effect of menstrual cycle on Doppler measurements of adnexa tumors]. AB - In a collective of 80 premenopausal women presenting with a total of 83 adnexal masses we tried to find out whether there is any influence on Doppler results by the phase of menstrual cycle. All measurements were performed with a 5.0 MHz transvaginal probe of an Acuson 125 Xp10 between April 1993 and September 1998. In 39 tumours measurements were performed during the follicular phase, in 44 tumours during the luteal phase. Histopathological evaluation showed 72 benign and 11 malignant adnexal masses. A cut-off of 0.69 for PI and 0.45 for RI was used to differentiate preoperatively between benign and malignant masses. In the follicular phase was a sensitivity of 40% and a specificity of 76.5% for RI. Referring values for PI were 60% and 70.6%. In the luteal phase RI led to a sensitivity of 83.3% and a specificity of 71.1%. Regarding PI a sensitivity of 100% and a specificity of 68.4% was achieved. There was no statistically significant difference when sensitivity and specificity for both parameters were compared in the two phases of menstrual cycle. These results are in contrast to the hypothesis that Corpora lutea are responsible for a higher degree of false positive results during the luteal phase. We therefore think that there is no advantage in performing Doppler measurements strictly during the follicular phase. PMID- 10795118 TI - [Dose intensified adjuvant chemotherapy in high risk breast carcinoma with 4-9 positive lymph nodes]. AB - OBJECTIVE: Taxanes and anthracyclines represent the two most active groups of agents for the treatment of breast cancer. We evaluated this combination in patients with more than 3 positive lymph nodes in an adjuvant, dose-intensive, sequential therapy in comparison with the standard chemotherapy regimen epirubicin/cyclophosphamide in relation to toxicities. MATERIAL AND METHODS: Since 9/96 127 patients with 4-9/over 9 positive lymph nodes have been recruited from 21 participating centers in an ongoing trial. 67 patients were prospectively randomised for first-line chemotherapy to treatment group A (epirubicin 90 mg/m2 paclitaxel 175 mg/m2; 4 cycles bi-weekly, supported by G-CSF 5 micrograms/kg day 5-13 and 3 sequential cycles of CMF 600/40/600 mg/m2 at 2-weeks interval) and 60 patients to treatment group B (epirubicin 90 mg/m2-cyclophosphamide 600 mg/m2, 4 cycles tri-weekly, and 3 sequential cycles of CMF 600/40/600 mg/m2 at 3-weeks interval). RESULTS: Preliminary safety and toxicity data are evaluable for 679 cycles. Data about response rate and disease-free-survival and overall survival will be delivered later. For the hematological toxicity the main grade 3 and 4 adverse events for A vs. B were: leucopenia 9.8% vs. 8.4%, febrile neutropenia 1.6% vs. 0.8%--anemia (< 5.9 mmol/l), 0.4% vs. 0.2%--thrombopenia 0% vs. 0%. Non hematological toxicity occurred more frequently in group A (grade 2, 3, 4):- neuropathy 4.4% vs. 0%,--nausea/emesis 27.8% vs. 19.3%,--fatigue 14.6% vs. 3.4% and mucositis 2.8% vs. 0.3%. PMID- 10795119 TI - [Maternal CA 125 serum level in intrauterine pregnancy and abortion in the first trimester]. AB - OBJECTIVE: Maternal CA 125 levels are supposed to rise in pregnancies complicated by vaginal bleedings in dependence to the extent of decidual disruption which is directly related to the outcome of pregnancy. MATERIAL AND METHODS: The prognostic value of maternal CA 125 serum measurement was investigated in 239 women with a first trimester intact pregnancy, imminent, incomplete, complete or missed abortion. RESULTS: 43.9% of the CA 125 serum levels were without normal range (> 20 U/ml). Mean CA 125 serum levels were higher in patients with incomplete (52.4 +/- 67.4 U/ml), complete (34.3 +/- 46.1 U/ml), and imminent abortion (33.0 +/- 45.8 U/ml) as compared with normal pregnancies (28.9 +/- 28.8 U/ml) and missed abortion (23.5 +/- 21.5 U/ml). CA 125 levels in first trimester pregnancies tended to be higher in patients with vaginal bleedings than in patients without bleeding (40.5 U/ml +/- 55.0 vs. 28.9 U/ml +/- 28.8; p = 0.65). CONCLUSIONS: For clinical use CA 125 serum measurement is not relevant. First trimester CA 125 measurement can not serve as an accurate predictor of pregnancy outcome due to the wide overlap of ranges. PMID- 10795120 TI - [Room air as the etiology of gas embolism in diagnostic CO2 hysteroscopy]. AB - During a diagnostic CO2-hysteroscopy in general anesthesia, a manifest gas embolism with a resulting drop of the endexpiratory CO2 partial pressure occurred upon insertion of the instrument. By ending the procedure and through appropriate anesthesiological measures, the occurrence was brought under control and the embolism had no clinical consequences. The incidence encouraged us to reconsider the CO2-hysteroscopy examination technique. As a result, we describe an up to now neglected mechanism which may lead to air embolism in gas hysteroscopy: Similar to hysteroscopy with fluid distension, the whole system has to be purged from air by insufflating CO2 prior to examination. If this step is neglected, up to 40 cm3 of room air may be insufflated into the patient, considering a connective tubing of 200 cm length and 0.5 cm lumen. The scientific organisations as well as the endoscopic training centers and the manufacturers of hysteroflators are challenged to deal with this newly described potential cause of complications. PMID- 10795121 TI - [Gynecological and obstetric discussion forums on the internet]. AB - The number of patients, especially with higher education, participating in medical discussion forums, news-groups and chats has increased rapidly during the past years. Physicians should be cognizant of this activity, for it alters the patient's attitude towards the doctor. Medical professionals themselves participate to a much lower degree in medical internet activities. PMID- 10795122 TI - [Information processing in the gynecological endocrinological laboratory. Current concepts and integration strategies]. AB - While computer applications are widely accepted in clinical chemical departments, small numbers of computer systems are used in gynecological endocrinological laboratories. Modern applications should be integrated in local area networks and clinical information systems that improve communication and data exchange within hospitals and the outer resources. Better communication may improve medical quality and will lead to a better therapy for gynecological endocrinological patients. PMID- 10795123 TI - Workplace managed care: collaboration for substance abuse prevention. AB - This article describes the history, purpose, and overall methodology of the Workplace Managed Care (WMC) study sponsored by the Substance Abuse and Mental Health Services Administration, Center for Substance Abuse Prevention (CSAP). This study was initiated to discern best practices for workplaces and managed care organizations integrating their substance abuse prevention and early intervention programs, strategies, and activities for employees and their families. CSAP funded nine WMC grants to study their retrospective and prospective data. Results of the WMC study suggested the addition of substance abuse prevention material to existing workplace health promotion offerings that resulted in improved substance abuse attitudes without jeopardizing existing health promotion programs. Stress management programming was successful at improving substance abuse attitudes indirectly. This study provides a platform for multidisciplinary research in workplace and managed care settings. PMID- 10795124 TI - Substance abuse intervention for health care workers: a preliminary report. AB - The Workplace Managed Care Cooperative Agreement project targets 3,300 health care professionals in hospital, specialty clinic, and primary care settings located in metropolitan New Mexico communities. This project will evaluate whether enhancements to existing substance abuse prevention/early intervention programs can prevent the onset of risky drinking, reduce prevalence of risky drinking, better identify employees who abuse alcohol and drugs, and improve employee wellness. This article describes one such enhancement (Project WISE [Workplace Initiative in Substance Education]), implemented at Lovelace Health Systems. Project WISE includes relatively low-cost elements such as substance abuse awareness training, information on how to reduce drinking, and brief motivational counseling. Evaluation will consist of baseline comparisons of the intervention and comparison sites, a process evaluation, a qualitative analysis using focus groups, and an outcome evaluation using health and work records. Methodological challenges, solutions, and implications for researchers undertaking similar projects are presented. PMID- 10795125 TI - Methodological issues in workplace substance abuse prevention research. AB - Substance abuse among working adults represents billions of dollars in preventable health care costs and industry financial loss. Therefore, it is imperative to develop and test effective substance abuse prevention programs for the workplace. However, applied workplace substance abuse prevention research is fraught with numerous methodological challenges. This article highlights a number of these challenges, which include (1) reaching a broad audience with prevention messages, (2) handling the concerns of the employer, (3) collecting substance use data in the workplace, (4) accessing and using records-based data, and (5) linking survey and records-based data. Using examples from the authors' ongoing research assessing a workplace health promotion and substance abuse prevention program, funded by the Substance Abuse and Mental Health Services Administration's Center for Substance Abuse Prevention, the authors address these challenges. PMID- 10795126 TI - The Employee Stress and Alcohol Project: the development of a computer-based alcohol abuse prevention program for employees. AB - The Employee Stress and Alcohol Project (ESAP) developed an interactive computer based alcohol abuse prevention and early intervention program accessible to employees over the Internet. Behavioral health research recommends that specialists develop and provide comprehensive yet cost-effective approaches to alcohol abuse prevention, early intervention, and treatment within the context of workplace managed care. ESAP is implementing this web site for a diverse 8,567 employee work site. ESAP's web site enables employees to self-assess their stress levels, coping styles, and risk for alcohol-related problems. It provides personalized feedback, recommendations, mini-workshops, a drinking journal, links to other online resources, and an interactive forum for direct participant-to participant communication. ESAP's web site provides resources for employees who are concerned about another individual's drinking as well as information about adolescent alcohol use. This article discusses the implications of incorporating the ESAP web site into worksite alcohol abuse prevention and early intervention programming. PMID- 10795127 TI - Methods for evaluating a mature substance abuse prevention/early intervention program. AB - The authors describe methods for work in progress to evaluate four workplace prevention and/or early intervention programs designed to change occupational norms and reduce substance abuse at a major U.S. transportation company. The four programs are an employee assistance program, random drug testing, managed behavioral health care, and a peer-led intervention program. An elaborate mixed methods evaluation combines data collection and analysis techniques from several traditions. A process-improvement evaluation focuses on the peer-led component to describe its evolution, document the implementation process for those interested in replicating it, and provide information for program improvement. An outcome assessment evaluation examines impacts of the four programs on job performance measures (e.g., absenteeism, turnover, injury, and disability rates) and includes a cost-offset and employer cost-savings analysis. Issues related to using archival data, combining qualitative and quantitative designs, and working in a corporate environment are discussed. PMID- 10795128 TI - Why carve out? Determinants of behavioral health contracting choice among large U.S. employers. AB - Many U.S. employers have carved substance abuse and mental health services out of their medical plans, changing the way millions of people access care. Employers that take this approach contract directly with specialized vendors, bypassing their general health plans. Since carving out may alter access and treatment, there is a need to understand why employers take this approach. This article reviews various hypotheses about why purchasers carve out and tests them using data from a survey of America's largest employers, the Fortune 500 firms. Size is the strongest predictor of an employer's decision to carve out behavioral health once other characteristics are controlled for. Employers that report they value coordination are less likely to carve out, while those that value special expertise are more likely to carve out. Employers are less likely to carve out enrollees in health maintenance organizations (HMOs) than those in other types of plans. PMID- 10795129 TI - Provision of drug treatment services in the juvenile justice system: a system reform. AB - This article proposes a systemic reform of the organizational structure and delivery of substance abuse services for adolescents within the juvenile justice system. It first discusses the impact of substance use on the juvenile justice system and then reviews which drug treatment programs and services are currently available. Following an evaluation of the most effective drug treatment programs and modalities, recommendations for system reform are given. The recommendations are based on a graduated sanctions framework, supported by systems collaboration and comprehensive case management. Systems collaboration between service providers must exist for juveniles to receive appropriate and comprehensive services. Case managers (CMs) both assess juveniles and help them move through and between judicial, drug treatment, and social service systems. In this way, juveniles receive the most suitable and complete services a community can offer while remaining firmly under juvenile justice system supervision. PMID- 10795130 TI - Managed care and fee discounts in psychiatry: new evidence. AB - This article describes the extent of managed care and fee discounting in psychiatric practice using data on 970 randomly sampled American Psychiatric Association members from the 1996 National Survey of Psychiatric Practice. Seventy percent of psychiatrists were found to have some patients in managed behavioral health care programs. The survey data illustrate that psychiatrists' involvement in managed care spans primary practice settings and is fairly evenly distributed across regions of the United States. Nationally, psychiatrists discount fees for 35% of their patients, with significant variation by practice type and extent of involvement in managed behavioral health care. The average level of discount is 25% with little variation by practice type or extent of involvement in managed behavioral health care. There is little evidence that psychiatrists with patients in managed care have higher fee levels than psychiatrists with no patients in managed care. PMID- 10795132 TI - [Not a taboo any more--but still impossible. Sexual dreams of the elderly. 4th Gutersloher Gerontopsychiatric Symposium]. PMID- 10795131 TI - Criminal behavior and emotional disorder: comparing youth served by the mental health and juvenile justice systems. AB - This study explored whether youth involved in joint service systems differed from single-agency users in terms of types of crimes committed and clinical functioning. Data from 4,924 youth involved in one county's public mental health and juvenile justice service systems were examined. Twenty percent of those youth receiving mental health services had recent arrest records, and 30% of youth arrested received mental health services. Of all youth arrested in the county, mental health service users had more arrests than non-mental health service users. A subsample of 94 mental health service users with arrests was matched on demographics with 94 mental health service users without arrests. Youth with arrests had a higher frequency of conduct disorder, higher Child Behavior Checklist Externalizing and Total Problem Scale scores, and more functional impairment on the Child and Adolescent Functional Assessment Scale as compared to youth without arrests. Implications for behavioral health service delivery were discussed. PMID- 10795133 TI - [Per e-mail information about scientific progress. Medical updates to the house for free]. PMID- 10795134 TI - [Magic numbers]. PMID- 10795135 TI - [The surgeon was also thinking]. PMID- 10795136 TI - [Saving reimbursements with psychotherapy?]. PMID- 10795137 TI - [When the patient wakes up during surgery. Help for anesthesia victims. Interview by Dr. Brigitte Moreano]. PMID- 10795138 TI - [Hepatitis C. Saving the liver]. PMID- 10795140 TI - [In Process Citation] PMID- 10795139 TI - [Even in exercise-induced asthma, patients need not be unnecessarily protected!]. PMID- 10795141 TI - [Diagnosis of hypertension without residual doubt. Can increased blood pressure be reliably recognized in general practice?]. PMID- 10795142 TI - [Fat control--an effective antihypertensive strategy. Special recommendations for therapy of the overweight patient]. AB - Some 44% of all patients with elevated blood pressure are overweight. In obesity related hypertension, sympathicotonia is regularly found, together with elevated intracellular calcium, sodium retention, increased cardiac output (per minute) and a sensitivity to salt. The role played by hyperinsulinemia has apparently been overstated. A primary rise in the minimal vascular resistance suffices to reduce the perfusion of the skeletal musculature and, solely on this basis, to induce insulin resistance. For the treatment of obesity-related hypertension, non medicinal approaches to weight reduction predominate. Reducing the daily salt intake to 5 g can also bring about a measurable reduction in blood pressure. For the treatment with antihypertensive drugs, beta blockers and diuretics are the initial choice; in the case of pronounced metabolic syndrome, ACE-inhibitors and alpha-1 receptors. PMID- 10795143 TI - [Are differences between AT1 antagonists clinically relevant? Dosage, selectivity, metabolism--critically evaluated]. PMID- 10795144 TI - [Chronic renal failure--taming the sympathetic nervous system! New approach to delaying progression]. AB - Previous research in end stage renal failure has focused on the renin-angiotensin system. Today, we know, that in chronic renal failure sympathetic overactivity plays a pivotal role for progression of the disease and overall prognosis. The unique finding is that the damaged kidneys send afferent nerve impulses to the central nervous system to increase efferent sympathetic discharge. Systemically this sympathetic overactivity contributes to hypertension and associated cardiovascular complications. Locally in the kidney neurotransmitter release is enhanced, which induces proliferation thereby promoting loss of renal function. Consequently, it has been shown that pharmacological inhibition of sympathetic nerve activity prevents glomerulosclerosis and ameliorates proteinuria in models of chronic renal failure. Thus, the pathomechanism of sympathetic overactivity is likely to open a new therapeutic avenue for the treatment of hypertensive chronic renal failure patients. PMID- 10795145 TI - [New therapeutic recommendations. Lowering blood pressure--but evidence-based, please]. PMID- 10795146 TI - [Nightshade plants act almost like LSD. Poisoning cases are on the rise]. AB - Intoxicated states can be induced not only by the typical narcotic drugs, but also by other psychotropic substances, such as the thorn-apple and angel's trumpet. In recent times there has been an increase in the reported number of cases of such intoxications. Clinical symptoms, toxicological analysis and possible forms of treatment are discussed. PMID- 10795147 TI - [What do general practitioners expect from a psychiatric clinic? Closer cooperation is requested]. PMID- 10795148 TI - [Counseling before and during pregnancy. 4: Pregnancy in liver diseases and after liver and heart transplantation]. PMID- 10795150 TI - [Fischer loses control--Dressler triumphs. War of the roses around health care reform]. PMID- 10795149 TI - [Diagnostic quiz. Dyspnea and bone pain. Metastatic osteolytic-osteosclerotic osteosarcoma with pleural metastases]. PMID- 10795151 TI - [Hamburg KV complains. Social insurance is targeting us]. PMID- 10795152 TI - [License fee for fibromyalgia? Illness with trademark protection]. PMID- 10795153 TI - [East German nostalgia in Brandenburg? Comeback of polyclinics]. PMID- 10795154 TI - [A new procedure improves microcirculation. Controlled fibrinogen depletion should save diabetic feet]. PMID- 10795155 TI - [Inflammatory response in the allergic patient. Lemonade taste improves compliance]. PMID- 10795156 TI - [Prevention of embolism in severely ill internal medicine patients. In heart failure it is better to use low molecular weight heparin]. PMID- 10795157 TI - [Report of experiences. Established prognosis--but pain under control]. PMID- 10795158 TI - [Influenza loses it's fright]. PMID- 10795159 TI - [The last great epidemic of humanity]. PMID- 10795160 TI - [Examining light. On the origins of clinical diagnosis]. PMID- 10795161 TI - [How aggressive can sleepwalkers become? Motor acts of murder]. PMID- 10795162 TI - [Does the "Lion Heart" make donor hearts superfluous?. Interview by Dr. Ch. Berndt]. PMID- 10795163 TI - [Etiology and therapy of chronic fatigue syndrome. Too tired for life. Press Conference: Fatigue--Paralyzing Symptom in Cancer Patients, Cologne, 17 September 1999]. PMID- 10795164 TI - [Decubitus ulcer. Basic treatment intervention is pressure unloading]. AB - With an incidence of between 3 and 34%, decubitus ulcers are common chronic wounds, many of which can be avoided by prophylactic measures. The most effective preventive measure is relieving pressure on the endangered part of the body, and this is most easily achieved by regularly changing the patient's position in bed. Since, however, this is not always possible for staff-shortage or illness-related (e.g. fractures of the spine) reasons, modern pressure-relieving systems are being increasingly used. The range of options extends from simple foam plastic underlays to water-filled cushions to pneumatic cushions or beds filled with tiny glass beads. Selection of the most appropriate system is often difficult. For effective prophylaxis, determination of the individual risk of developing a bedsore with the aid of special scales makes good sense. In this way, the measures required can be adapted to the particular needs of the individual patient. New approaches to decubitus ulcer prevention and wound management may help to ensure effective care of the endangered or affected patient. PMID- 10795165 TI - [Don't let it get that far! Tips and tricks for prevention of decubitus ulcer and therapy in general practice]. AB - Successful prevention and treatment of decubitus ulcer requires a high level of skill and competence on the part of the diverse professional groups involved in providing nursing care and medical treatment. Pressure relief can be achieved in a variety of ways, and the needs of the patients must be integrated in the approach adopted. Constant inspection and care of the skin, and promotion of patient movement/activity, coupled with familiarity with the latest information on wound and pain management are the prerequisites for appropriate treatment. A reduction of the problem to the use of special mattresses and beds must be avoided. PMID- 10795166 TI - [Asthma--compact seminars for the general practitioner]. PMID- 10795167 TI - [Alternative medicine sails under a false flag. Alternative practitioners profit from the good reputation of naturopathy]. PMID- 10795168 TI - [New therapy procedure for localized cartilage defects. Encouraging results with autologous chondrocyte implantation]. AB - Owing to the poor regenerative capacity of cartilage, cartilaginous defects are considered to represent pre-arthrotic factors. In addition to autologous and allogenic osteochondral fragments, proliferative tissue, such as periosteum and perichondrium are increasingly being used as graft material. The aim of treatment is to eliminate the defect and to restore the load-bearing capacity and function of the affected joint. A new, recently introduced, approach aims to stimulate the formation of new cartilage via autologous cultured chondrocyte implantation (ACI). The rationale for this treatment is the restoration of loadable hyaline or hyaline-like articular cartilage. Although long-term results are not yet available, clinical follow-up data obtained so far are encouraging. In addition to existing methods of treating cartilaginous defects, this article describes a modified method of transplantation of autologous chondrocytes. With this method the periosteal flap used to cover a defect is replaced by an absorbable collagenl/III membrane (Chondrogide, Geistlich Wolhusen, Switzerland) that is used as a carrier for the patient's own chondrocytes. After placement in the defect, the membrane is fixed in place with fibrin glue (MACI). PMID- 10795170 TI - [Counseling before and in pregnancy. 3: Disorders of carbohydrate metabolism and thyroid gland function]. PMID- 10795169 TI - [Thrombophilia--often a case of genetic susceptibility. Thrombosis ABC, 6: Protein C deficiency]. PMID- 10795171 TI - [Stomatitis and itchy efflorescences. Lichen planus]. PMID- 10795172 TI - Meningococcal disease, Ethiopia (update). PMID- 10795173 TI - Measles outbreak, Netherlands. PMID- 10795174 TI - Schistosomiasis and soil-transmitted helminth infections. PMID- 10795175 TI - Application of modified CHIP-AE in a vocational high school. AB - Adolescent health behaviors tend to cluster and vary by such factors as grade, age, and racial/ethnic background. A population-based instrument was needed to assess health behaviors of the students in a vocational high school in the City of Chicago. To reduce respondent burden, Child Health and Illness Profile- Adolescent Edition (CHIP-AE) was modified by omitting seven subdomains. Ninety three boys and 245 girls voluntarily participated in the study. Pearson correlation coefficients, MANOVA, and ANOVA were used in data analysis. Modified CHIP-AE was found to have acceptable reliability and detect the expected differences of adolescent health behaviors by gender and age. Modified CHIP-AE can be used to assess adolescent health behaviors in an urban vocational high school. It can also be used to target the subgroups of students with specific needs for health services. PMID- 10795176 TI - Evaluation of a program to promote diabetes care via existing agencies in African American communities. AB - Some African Americans with (or at risk for) diabetes underutilize health care services. We report short-term results of a "training of trainers" workshop designed to address this problem. The training program includes culturally sensitive educational materials, including materials developed for the ADA's African American Program (AAP). Workshops were presented to a) the 1996 national meeting of the predominantly black National Missionary Baptist Convention's Nurses Guilds, b) a minority-owned, TN based managed care organization's "community outreach workers," and c) other interested community organizations. Evaluations were based on program satisfaction and an "intention to change" procedure that assessed participants' actions and the obstacles they faced 6 months later. Sixty-four group representatives from 13 states participated. They completed a satisfaction questionnaire and were asked to complete a form that asked them to check any of 12 diabetes-related actions (distributing ADA risk tests, offering AAP classes, etc.) they intended their church/community group to take within six months. Activities not listed could be added. Follow-up contact information was solicited. Satisfaction surveys were positive. 39 (61%) returned checklists with complete contact information. Intentions included: arrange for congregation/community group to take risk test (71% of respondents), distribute diabetes materials at community health fairs or church services (67%), present AAP modules (59%), promote healthy foods at pot luck suppers (56%) and arrange cooking or exercise classes (38%). Respondents were contacted by telephone 6 months post-workshop and asked whether they had fulfilled their intentions. Contact information for 6 (15%) was no longer valid, and we were unable to reach 7 others despite repeated attempts. Approximately 30% of intentions were fulfilled by nurses guild members, but less than 10% by other groups. Half of all fulfilled intentions occurred in a community served by an active ADA AAP Coalition. Barriers to fulfilling intentions included lack of time/support, group not ready to act or doing other programs, and failure to collaborate with the ADA or others for mutual assistance. Existing agencies, especially churches with nurses guilds, offer a means for promoting diabetes screening and awareness in African American communities. A training workshop was well received and influenced some participant groups' self-reported actions. Participants appear more likely to fulfill intentions to conduct diabetes-related programs when they collaborate with other churches, agencies and/or the ADA. PMID- 10795177 TI - A visit to the Republic of South Africa and Botswana: a modern country with a relic from the past. AB - In a global health and communication focused society, it is evident that there is a growing need for health care professionals, especially nurses, to face challenges beyond traditional nursing care. Cultural awareness and values affect all areas of a person's life, including their beliefs about health and illness. The recent trip to South Africa and Botswana, a land that seemed so far away, was just only 18 short airline hours within reach. The Association of Black Nursing Faculty, Inc., Chi Eta Phi Sorority, Inc., and the Department of Nursing Education (University of Botswana) sponsored the trip to engage in professional dialogue of health concerns of many disadvantaged populations around the world for the 21st century. Although, this dialogue was an assurance that nurses around the globe do face challenges beyond a traditional nursing practice, it was also an assurance that there was a receptiveness toward teaching and the willingness to engage in health promotion behavior within a dominant culture that influences the lives of many people. In a land of sunshine and scenic majesty, were some of the world's strangest and most dramatic landscapes, nature's richest treasure chest of gold and diamonds with a unique wealth of animal and plant life. This land is also the home of big game, a host of birds, and has a legacy that is one of the most scenically dramatic, healthiest and challenging regions of the world. PMID- 10795178 TI - Evolutions in coronary care. PMID- 10795179 TI - Management of coronary artery disease. PMID- 10795180 TI - Assessment of pain and agitation in critically ill infants. AB - Critically ill infants are subjected to many painful experiences that, if inadequately treated, can have severe physiological and psychological consequences. Optimal management of pain relies on the adequacy of nurses' assessment; this, however, is complicated by another common condition, agitation. A multidimensional assessment is therefore necessary to adequately identify pain and agitation. The aim of this descriptive study was to identify the cues that nurses caring for critically ill infants use to assess pain and agitation. A questionnaire, developed from the literature, was distributed to all registered nurses (85) working in the neonatal and paediatric intensive care units of an Australian teaching hospital. Questionnaires were completed by 41 nurses (a 57 per cent response rate). Results revealed that, except for diagnosis, there were no significant differences between the cues participants used to assess pain and those to assess agitation. Nurses used numerous cues from various sources: most importantly, their own judgement (99 per cent); the parents' judgement (90 per cent); the infant's environment; documentation (78 per cent), and the infant's cues (70 per cent). These findings demonstrate the relevance of the nurse's role in assessment of pain and agitation in critically ill infants. Nurses used cues specific to the critically ill rather than the less sick infant. Results of this study also show the difficulty of differentiating between pain and agitation. Further research on ways of distinguishing between the construct of pain and agitation needs to be undertaken. PMID- 10795182 TI - Liquid ventilation: a future modality? AB - Liquid ventilation, an idea currently being trialled in the United States, is increasingly being discussed as a possible future trend in ventilation. A review of the available literature indicates that this treatment provides effective gas exchange and has a number of potential advantages. These include lower airway pressures, decreased alveolar surface tension, alveolar recruitment and removal of pulmonary exudate. While yet to be seen in this country, liquid ventilation may be introduced in the future. If it is, those caring for patients treated in that way will require knowledge of the mechanics and physiological changes involved, as well as the potential hazards of this modality. PMID- 10795181 TI - Taking on the student role: how can we improve the experience of registered nurses returning to study? AB - Regardless of the number of nurses who undertake graduate certificate courses in critical care nursing in Victoria, Australia, there is still a shortage of skilled nurses in the critical care areas. Preparation for practice in this specialist field has moved from hospital-based programs to tertiary-based courses during the past decade. As a consequence, there has been scrutiny of the clinical ability of graduates to perform the role function. Although this is understandable, anecdotal evidence to date indicates that students in these courses experience many difficulties, yet very little is written about the stressors they encounter. As the future of the critical care work force depends on a constant supply of students undertaking courses, it is essential to gain insight into the student experience, with a view to optimising it for future students and perhaps preventing attrition from courses. This paper describes a qualitative research study involving 14 students undertaking a graduate certificate course in critical care nursing. The aim of the study was to document the students' experience in relation to factors they identified as helping or hindering their progress on the course. Data were obtained at three points during the academic component of the course: on entry, at the end of semester 1 and at the end of semester 2. Interviews and questionnaires were used as the data collection methods. Results indicate that for this group of students, the experience of undertaking the course was adversely affected by stress related to three factors: change of role from clinician back to student; unmet expectations, and exhaustion. Recommendations related to preparation for study, curriculum timelines and peer support are included in this paper. PMID- 10795183 TI - A review of intensive care nurse staffing practices overseas: what lessons for Australia? AB - In view of market-driven health-care policies and the move to greater efficiencies within the health-care system, the cost of nursing care is being increasingly scrutinised. Different overseas practices are commonly cited as justification for changing practices within Australia. This study is based on a review of the literature on intensive care nurse staffing requirements in Australasia; specifically, New South Wales, the United States (US) and, to a lesser extent, Europe. It was found that looking to the US for cost-cutting strategies in intensive care units (ICUs) is based on a false premise: that we are comparing like with like. ICUs in the US have a different historical trajectory and culture, service wider constituencies, have technicians and unregistered personnel providing nursing care and do not provide demonstrably better outcomes or significant cost savings. Research indicates that continuous nursing care by trained professionals provides the best outcomes. If costs must be cut, technology, pharmaceuticals and laboratory tests should be targeted. Further, a greater commitment to the development of a 'progressive patient care' model in hospital planning is required, in order to establish or consolidate an intermediate level of nursing care between the ward and the ICU. Programs aiming to improve and continuously monitor patient care, such as adverse event monitoring, the prevention of unplanned extubation and facilitation of early extubation, should be instituted, as these have been shown to not only reduce ICU costs but also improve patient outcomes. PMID- 10795184 TI - Survivors of acute respiratory distress syndrome may experience lower quality of life than other populations. PMID- 10795185 TI - The future in nursing is genetics. PMID- 10795186 TI - Parenting education. Implications for research, practice, and teaching. PMID- 10795187 TI - Infant arousal in an en face exchange with a new partner: effects of prematurity and perinatal biological risk. PMID- 10795188 TI - Perceptions of secondary school staff toward the implementation of school-based activities to prevent weight-related disorders: a needs assessment. PMID- 10795189 TI - The Juvenile Wellness and Health Survey (JWHS-76): a school-based screening instrument for general and mental health in high school students. PMID- 10795190 TI - Availability of day care services for preschool children with special health care needs. PMID- 10795191 TI - Predicting adolescent peer problems and antisocial activities: the relative roles of aggression and dysregulation. PMID- 10795192 TI - Potential for abusive parenting by rural mothers with low-birth-weight children. PMID- 10795193 TI - Is there a role for pediatric nurses in the evolving field of human genetics? PMID- 10795194 TI - I look funny and I feel bad. Measurement of symptom distress. PMID- 10795195 TI - Genetics on the Internet! PMID- 10795197 TI - A meeting of ophthalmology minds from North and South America. PMID- 10795196 TI - Promoting safe and effective genetic testing in the United States. Final report of the Task Force on Genetic Testing. PMID- 10795198 TI - Commonly asked questions about diabetic retinopathy. PMID- 10795199 TI - Introduction to formal visual field testing, Part II: Anatomic correlation to visual field abnormalities. PMID- 10795200 TI - Applanation tonometry. PMID- 10795202 TI - Update on contact lens solutions. PMID- 10795201 TI - Communicating with laser vision correction patients. AB - Laser vision correction or LASIK has a long and excellent history with significant benefits and unlikely risks. Adequately educating patients should allay their fears and allow good candidates to enjoy the life changing benefits of improvement in the natural vision through laser vision correction. PMID- 10795203 TI - Listen to the children. PMID- 10795204 TI - Thoughts on theory. PMID- 10795205 TI - JCAHO accreditation considerations for the home care WOC nurse. AB - Driven by competition and by pressure from managed care groups, home health care organizations are increasingly choosing to seek the accreditation offered by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO). WOC nurses practice in a variety of health care settings, including home health care. Many home health care organizations are now accredited, or they will choose to become accredited in the future. This article reviews home care accreditation standards as set forth by JCAHO and the possible impact of the standards on individual WOC home care nursing practice. PMID- 10795206 TI - Using wound care algorithms: a content validation study. AB - PURPOSE: Valid and reliable heuristic devices facilitating optimal wound care are lacking. The objectives of this study were to establish content validation data for a set of wound care algorithms, to identify their associated strengths and weaknesses, and to gain insight into the wound care decision-making process. METHODS: Forty-four registered nurse wound care experts were surveyed and interviewed at national and regional educational meetings. Using a cross sectional study design and an 83-item, 4-point Likert-type scale, this purposive sample was asked to quantify the degree of validity of the algorithms' decisions and components. Participants' comments were tape-recorded, transcribed, and themes were derived. RESULTS: On a scale of 1 to 4, the mean score of the entire instrument was 3.47 (SD +/- 0.87), the instrument's Content Validity Index was 0.86, and the individual Content Validity Index of 34 of 44 participants was > 0.8. Item scores were lower for those related to packing deep wounds (P < .001). No other significant differences were observed. Qualitative data analysis revealed themes of difficulty associated with wound assessment and care issues, that is, the absence of valid and reliable definitions. CONCLUSION: The wound care algorithms studied proved valid. However, the lack of valid and reliable wound assessment and care definitions hinders optimal use of these instruments. Further research documenting their clinical use is warranted. Research-based practice recommendations should direct the development of future valid and reliable algorithms designed to help nurses provide optimal wound care. PMID- 10795207 TI - Validity and reliability: basic research concepts. PMID- 10795209 TI - Developing and implementing a wound care program in long-term care. AB - Pressure ulcers in residents of long-term care facilities cause avoidable pain and suffering, and they place an additional burden on budgets that are already quite lean. To laypersons, these "bedsores" inaccurately enhance a widely held belief that nursing homes provide inferior care. Although most long-term care administrators recognize the need for better prevention and management of pressure ulcers, they lack specialized knowledge in the field of wound prevention and treatment and lack the necessary time to devote to such an effort. A systematic approach using prevention and treatment protocols and a wound care team and staff education, has proved highly effective in reducing the incidence of pressure ulcerations. This article provides a detailed guide for the development and implementation of a wound prevention and care program and defines the critical role played by the WOC nurse within such a program. PMID- 10795208 TI - Techniques for applying subatmospheric pressure dressing to wounds in difficult regions of anatomy. AB - Subatmospheric pressure dressing (SPD) has been commercially available in the United States since 1995 as the vacuum-assisted closure (VAC) device. SPD increases local blood flow, decreases edema and bacterial count, and promotes the formation of granulation tissue. Despite recent clinical successes with the use of SPD in a variety of wound types, problems may occur with application of VAC system in certain areas of the body. The main limitation occurs when attempting to maintain an airtight seal over irregular surfaces surrounding a wound. For example, application of the adhesive drape and creation of a seal are particularly difficulty in the hip and perineum. In addition, wounds of the lower extremity can occur in multiple sites, posing the problem of providing a vacuum dressing to more than one wound from one suction pump machine. To address these challenging clinical wounds, we have developed techniques to allow the successful application of SPD to sacral pressure ulcers near the anus, and to multiple large lower extremity ulcers. PMID- 10795210 TI - Staff compliance with and ratings of effectiveness of a prompted voiding program in a long-term care facility. AB - BACKGROUND: Despite findings that prompted voiding is effective in reducing urinary incontinent (UI) episodes, the prevalence of UI in this population has remained unchanged. It is estimated that 50% of all nursing home residents have UI. Staff involvement is considered a critical factor, yet little is known about staff perception of the effectiveness of prompted voiding. METHODS: This descriptive study of staff perceptions of a prompted voiding intervention was conducted at a university-affiliated 255-bed geriatrics center. A 12-week prompted voiding program was conducted with 17 residents with UI. Baseline continence status was compared with continence status at the end of the program, and a survey was conducted to assess staff perceptions of the program. The aims of the study were: (1) to determine effectiveness of a prompted voiding program, (2) to assess staff perception of overall effectiveness of the prompted voiding program, (3) to assess staff compliance with the prompted voiding program, and (4) to compare staff perception of prompted voiding effectiveness with actual continence outcomes. RESULTS: Overall UI status improved in 5 residents (31%), remained the same in 6 residents (38%), and declined in 5 residents (31%). Sixty four (73%) of 88 staff members who participated in the program responded to a survey. Staff members were asked to rate their overall impressions of the prompted voiding program, and to rate improvement in continence status for specific residents who participated in the program. Ninety-five percent of staff members thought drier residents were happier; 80% thought the prompted voiding program should continue. Only 52% of staff thought residents participating in the prompted voiding program were better, and 43% perceived no change. A majority (58%) of staff thought compliance with prompted voiding was 80% to 90%; however, unobtrusive observations during the study revealed only 70% compliance. There was no correlation between staff ratings of improvement in continence and actual continence outcomes for individual residents (r = 0.02, P = .709). CONCLUSIONS: Staff reports of compliance with a prompted voiding program were inflated, and they were unable to determine which residents had actually improved and which residents had not improved with regard to UI. These findings suggest that nursing home staff, and particularly nursing assistants, need more meaningful definitions of UI and quantifiable evidence that residents benefit from prompted voiding. PMID- 10795211 TI - Urinary incontinence in a geriatric patient with a complex neurologic history. PMID- 10795212 TI - Management of extensive peristomal ulcers around a retracted, stenotic ileal conduit stoma site. PMID- 10795213 TI - Diagnostic imaging: an overview. PMID- 10795214 TI - Diagnostic testing for coronary artery disease. PMID- 10795215 TI - Abdominal pain protocol: right upper quadrant pain. PMID- 10795216 TI - Ultrasound, magnetic resonance imaging, and computed tomography. PMID- 10795217 TI - Radiology of abdominal pain. AB - Radiology plays an integral role in the evaluation of patients with significant abdominal pain. The cross-sectional modalities (computed tomography, ultrasound, and magnetic resonance imaging) are widely used, but there is sometimes confusion about how to use each test appropriately. We review how each test is done, consider the strengths and weaknesses of each modality, and discuss how to use them in an intelligent, cost-effective manner. PMID- 10795218 TI - Pearls of the plain film. AB - Three case presentations from a general rheumatologic practice show the importance of the plain x-ray in making the diagnosis of arthritis. Clinical history, physical examination, and the plain x-ray are likely to provide the diagnosis most often over laboratory and more technologically advanced imaging studies for arthritis. PMID- 10795219 TI - Radionuclide bone scanning in routine clinical practice. AB - Bone scintigraphy (scanning) is a quick and cost-effective method for evaluating potential bone lesions not visualized on plain radiography. Uses include evaluation of primary and metastatic bone tumors, treatment of bone pain due to tumor invasion, detection of occult fractures and stress injury, evaluation of postprosthesis joint pain, evaluation of metabolic bone disorders, and diagnosis of osteomyelitis. The main advantage of bone scanning is the ability to evaluate the entire skeleton rapidly with high sensitivity. PMID- 10795220 TI - National vocational qualifications. NVQs: its time they got the recognition they deserve. PMID- 10795221 TI - National vocational qualifications. Associate practitioners in the critical care unit. PMID- 10795222 TI - Clinical effectiveness. Implementing clinical effectiveness: a pragmatic approach. PMID- 10795223 TI - Critical appraisal. Finding and appraising cohort studies for causation and prognosis. PMID- 10795224 TI - Making sense of learning contracts. 1. Using a learning contract. PMID- 10795225 TI - Reflective practice. Making reflection a rewarding and productive experience. PMID- 10795226 TI - Professional regulation: the key to practice that is fit for purpose. PMID- 10795227 TI - Critical appraisal. 5. Randomised controlled trials: questions for valid evidence. PMID- 10795228 TI - Information technology. Open learning with Merlin. PMID- 10795229 TI - Clinical governance. 3. Effective clinical practice--1. PMID- 10795230 TI - Effective communication in teaching and learning. 1. Understanding how people learn. PMID- 10795231 TI - Action learning. A beginner's guide to the principles of action learning. PMID- 10795232 TI - Critical appraisal. How to decide if review articles are trustworthy and relevant for practice. PMID- 10795233 TI - Role transition. Supporting newly qualified nurses. PMID- 10795234 TI - Why the biological sciences should be back in the curriculum. PMID- 10795235 TI - Clinical governance. 4. Effective clinical practice--2. PMID- 10795236 TI - Effective communication in teaching and learning. 2. Understanding our agendas. PMID- 10795237 TI - Transformational leadership. PMID- 10795238 TI - Critical appraisal. 8. Questions for surveys. PMID- 10795239 TI - Nurse-teacher roles--1. Nurse teachers' dilemma on how to manage their split role. PMID- 10795240 TI - Clinical governance. 5. Effective staff--1. PMID- 10795241 TI - Making sense of learning contracts. 2. Creating the right learning climate. PMID- 10795243 TI - Renal teaching cards: a visual way to teach patients. PMID- 10795242 TI - Incorporating genetics into nursing practice. PMID- 10795244 TI - Using the Internet to enhance a pharmacology course. PMID- 10795245 TI - Summertime and the learning is easy: making summer classes come alive. PMID- 10795246 TI - Selected response test items. AB - Classroom assessment is complex and challenging. Teachers need to consider the cognitive, affective, and psychomotor levels for achievement of their educational objectives. This series of six articles discusses how to develop testing blue prints; selected-response tests, including multiple-choice, true-false, matching, or other objective tests; completion or essay testing; problem solving/critical thinking activities; performance assessment; and computer-based testing. PMID- 10795247 TI - Spotlight on ... seventeen ways to transform reading assignments into critical thinking experiences. PMID- 10795248 TI - Nightingale's resume. A student career-teaching strategy. PMID- 10795249 TI - Health promotion for farmers at the country market. PMID- 10795250 TI - Professor Plum at the podium: clues for teleteaching. PMID- 10795251 TI - An electronic approach to evaluating healthcare Web resources. AB - Information literacy skills, which include the ability to evaluate electronic healthcare sites, are critical to the decision-making responsibilities of students and professionals. The authors describe the experiences of nursing and library services faculty in the development and implementation of a Web-delivered module for the evaluation of healthcare Web resources. A range of electronic tools was used for both the collaborative creation of the module as well as the instructional delivery of the content. Evaluation of the module revealed its usefulness to the students and the potential of the collaborative development model for other content areas. PMID- 10795252 TI - Rural health content in nursing education. PMID- 10795253 TI - Teaching communication about human sexuality to nurses and other healthcare providers. AB - Nursing and healthcare professional faculty are challenged to prepare graduates who are comfortable and capable of discussing human sexuality issues and concerns with patients who are victims of family violence, AIDS and other sexually transmitted diseases, rape, and abuse, as well as patients who have undergone cardiac surgery and other life-threatening events. The authors recommend using a holistic approach as the basis for teaching strategies. PMID- 10795254 TI - When a student commits suicide. A protocol for colleges of nursing. AB - Nursing faculty are rarely prepared for the suicide of a student. The authors discuss a protocol to be implemented when such an event occurs. It highlights the needs of faculty, fellow students, and significant others during this difficult period and guides the reader through the process of informing the faculty and students about the suicide, conducting support meetings, and managing the media. The protocol helps schools prevent disorganization and unnecessary legal jeopardy associated with such a crisis. PMID- 10795255 TI - Clinical simulation laboratory. An adjunct to clinical teaching. AB - Videotaped and telephone simulations role-played by students were used in a clinical laboratory setting to enhance student learning. The authors discuss the structure, implementation, evaluation, and potential uses of these simulated activities. PMID- 10795256 TI - Using case studies to teach clinical problem-solving. AB - The case study method is a useful student-centered learning strategy that allows students to solve real-world problems in the safe environment of the classroom. The authors discuss the strengths and limitations of this method and strategies for developing and using case studies in the classroom. PMID- 10795257 TI - Cultural competence case scenarios for critical care nursing education. AB - Cultural competence is a part of professional nursing practice and should be integrated into each course of nursing education. In a rural Midwest community, the clinical opportunities for cultural encounters in critical care may be limited. One strategy to promote cultural competence in the critical care nursing class of a baccalaureate program was to assign narrative responses to case scenarios. These case scenarios were discussed in the context of cultural competence. PMID- 10795258 TI - Teaching epidemiology via poster session. PMID- 10795259 TI - Active learning strategies to teach case management. AB - Changes in healthcare delivery require a new understanding of the concepts of case management in managed care. The authors describe the construction and evaluation of a learning module that encourages active engagement and skill development. Students develop an understanding of how to plan for care of populations in an effort to meet managed care demands. This approach to teaching case management can be effective in an educational setting and possibly in clinical settings as well. PMID- 10795260 TI - Call me coach. PMID- 10795261 TI - The myths of acquaintance rape. PMID- 10795262 TI - One practice week at a glance. PMID- 10795263 TI - Whistle-blowing: effective and ineffective coping responses. AB - How do nurses respond when they identify misconduct in patient care settings? The authors used a descriptive survey (N = 95) to identify effective coping strategies of whistle-blowers (n = 70) and non-whistle-blowers (n = 25). Results identified four effective coping strategies used by whistle-blowers. The authors offer recommendations to nurses who encounter misconduct in their workplace. PMID- 10795264 TI - Referencing in transcultural nursing: an ethical analysis. AB - The increasing emphasis placed on transcultural care among clinicians and educators, coupled with the lack of original research in this field, has led to a number of articles in journals and books that are poorly researched and referenced. Authors have a responsibility to critically review primary sources rather than rely on secondary sources that can be erroneous. Having established a unique contribution to the transcultural literature, nurse authors and educators have a responsibility to maintain high professional and ethical standards when disseminating information to readers. PMID- 10795265 TI - Substance abuse disorders in nurses. AB - Substance abuse is a serious concern in the profession of nursing. The American Nurses Association (1997) estimates that 10% to 20% of nurses have substance abuse problems, and that 6% to 8% of registered nurses are impaired due to their abuse of alcohol and other drugs. Chemical dependency is considered a disease that requires treatment. Early identification and treatment of the chemically dependent nurse is important for the safety of the public and for the well-being of the nurse and her profession. This article addresses substance abuse from a biopsychosocial perspective, and includes a description of an approach to treatment and suggestions for the role of nursing administration. PMID- 10795266 TI - Are you an academic cowboy? PMID- 10795267 TI - Nursing, technology and the millennium. PMID- 10795268 TI - Embracing technology: an exploration of the effects of writing nursing. AB - Contemporary societal factors influencing the delivery of healthcare in Canada include loud and persistent demands for accountability around the use of public funds. Interests in fiscal accountability are not only held by managers and politicians. Practitioners too hold interests in this domain, particularly where 'interest' means having a reserved seat at the table where discussions regarding resource allocation will take place. In this light, practitioners are also interested in enrolling forms of technology that enable them to make visible their 'unique contribution' to the care delivery enterprise. Latour's writings on 'technology' as durability offer methods for describing socio-technical processes that contribute to the appearance of domination. Challenging the prevailing view that order results from powerful domination, Latour argues that domination is an effect of contingent processes. A case study is presented where multiple interests in developing a form of nursing that is visibly 'autonomous' and 'cost effective' are brought together with technological forms that promise such visibility. Costs associated with purchasing visibility are critically examined. PMID- 10795269 TI - Understanding technology in contemporary surgical nursing: a phenomenographic examination. AB - The purpose of this paper is to describe the result of phenomenographic research undertaken to identify the qualitatively different ways technology is understood and experienced by contemporary surgical nurses. Twenty surgical nurses were interviewed and conversations were recorded through the use of drawings and audiotape that were analysed using a series of seven analytical steps. There were two major outcomes of the research: eight categories of description were revealed and each one constitutes a conception of the experience and understanding of technology, and an outcome space portraying the logical relations between conceptions was identified. Conceptions revealed demonstrate the importance of technology to the theory, practice, research and education of nurses, and highlight the complexity and significance of technology in contemporary surgical nursing experience. PMID- 10795270 TI - Situating wound management: technoscience, dressings and 'other' skins. AB - This paper addresses the notion of wound care as a technology of skin and other skins imbued with the combined power of technology and science. It presents the discourses of wound care evident in the accounts of patients and nurses concerning this care, and discussions about wounds in wound care interest groups, journals, and advertising material about wound care products. The discussion focuses on wounds and wound dressings as effects immanent in the power relations of discourses of wound care. These effects colour and influence nurses' responses to wounds and wound care products. Moreover, the discourses that portray these practices are evidence of the complex articulation between technoscience and gender. Nurses and patients are fascinated by wound technoscience and lured towards it by its potential for mastery and control over wounds. Such seductions are evident in the texts of nurses, patients, and pharmaceutical advertisements for wound care products. Finally, the ways that these representations are used to talk about and market wound care products are shown as exemplifying the finer points of wound management as a nursing technoscience. PMID- 10795271 TI - Virtual power: gendering the nurse-technology relationship. AB - To date, studies of the relationship between technology and its consumers have used the constructs of traditional paradigms of production and consumption as the foundation for analysis. These studies have served to reinforce traditional concepts of gender and hierarchy in the nursing-technology dichotomy. To propose a new and more relevant framework for analysing the technology-nursing relationship, the analysis of gender within the methodology of the social history of technology will be used. Healthcare will be viewed as a technologic network, and within that network multiple knowledge domains reside and interact. These domains, in turn, are socially constructed and historically contingent. This paper operationalizes this argument by examining the domain of the early nurse practitioner movement of the 1960s as part of a gendered technologic system. The findings of this study illuminate the agency of nurses in the shaping of traditionally male knowledge domains and as a crucial factor for understanding the evolution of not only the particularities of the nurse-technology relationship, but also the generalities of the gendered ways of knowing within the healthcare-technology relationship. Perhaps most importantly, different sets of questions can be formulated to analyse the history of the nurse practitioner movement from a technologic perspective that will provide new standpoints for the nursing-technology dichotomy in the millennium. PMID- 10795272 TI - The problem of dissemination: evidence and ideology. AB - This paper recontextualises research evidence as an example of textually-based social control. It does this by drawing on two areas of theoretical literature; feminist literary theory and the sociology of scientific knowledge. Accounts of literary works as ideological instruments of social control suggest that (at least some kinds of) research literature may fulfil a similar role among a clinical readership. There have also been compelling accounts of scientific writing as expressions of desire on the part of one group to 'act at a distance' upon others. In the light of this literature, it becomes less tenable to see research dissemination as the simple transfer of information, supplemented by organisational work. Research is implicated in the attempt by one group to enrol others in its own project and in the (self-)construction of the identities of the healthcare worker. The accounts that literary theory can provide do not remain focused upon the text, but draw links between the reading process and the experience and place in society, for example the gender, of the writer and reader. As such their explanations create a space for the resisting reader. PMID- 10795273 TI - Troubling distinctions: a semiotics of the nursing/technology relationship. AB - I consider the discursive practices that have served conceptually and ontologically to trouble the boundaries between nursing and technology: between nurse/human/subject and machine/non-human/object. Nursing and technology have been semiotically related largely by two processes: (a) by the metaphor that depicts nursing as technology and (b) by opposition, or as not like and even in conflict with technology. Less frequently but no less significantly, nursing and technology have been semiotically linked (c) by the metaphor that depicts technology as nursing and (d) by metonymy, or by word or picture juxtapositions of nursing with technology. The troubling distinctions between nursing and technology suggest yet another reason why the construction of difference continues to elude nursing. PMID- 10795274 TI - Nursing older people: constructing need and care. AB - As the population of the world 'ages', older people are increasingly viewed as a 'problem' whose needs challenge the resources of care services. However, this view is predicated on the negative and nihilistic attitudes that are held throughout society about older people and the effectiveness of therapeutic intervention. It is these pessimistic views which colour professional understandings of the needs of older people and the most appropriate ways of delivering care to meet those needs. However, any knowledge of need and care is a constructed story, and is limited by the knowledge base on which it rests. For example, western healthcare is dominated by an orientation to identified problems, the minimizing of physical risk or harm, and a reductionalist approach to problem solving. A social critical challenge to prevailing understandings of need and care produces a very different story about older age; one which emphasizes an individual's strengths and their family and community relationships. In exploring these issues in policy and practice two key areas are highlighted. One is the role of critical gerontology in challenging the stories of negativity about old age, and replacing these with positive stories that allow older people to retain their social purpose. The second key area is the need to address issues of older age at both a practice and policy level, and to understand the interrelationship between them. PMID- 10795275 TI - A psycho for every generation. AB - For almost 40 years, thanks to Alfred Hitchcock's seminal film, the word Psycho has conjured up some of the strongest and most enduring, not to say frightening and even misleading images of mental illness in contemporary western culture. It has become a byword for terrifying homicidal impulses, the unpredictability of a deranged mind, and has created, in Norman Bates, an archetype for the Other. However, we must now accept that time has passed. There is now another Psycho with which we have to deal, so close in shot selection and script as to pass for identical. Gus van Sant has produced a Psycho for the 1990s--or so we might consider. This paper will examine the way in which the two films, one produced in 1960 and the second in 1988, deal with the notions of mental illness, madness and the construction of the Other. It will pay particular attention to the central character of Norman Bates, how and why he is so terrifying to us, how his madness is explained and demonstrated and what a deconstructed analysis of that portrayal may signal. It will consider the notion that, just as we have come to accept that every generation may produce its own interpretation of roles from the classical stage repertoire, such as Hamlet, every generation can and will interpret Psycho to mirror its own concerns. PMID- 10795276 TI - Exploring motherhood: when the researcher has multiple roles. PMID- 10795277 TI - India: my mother country. PMID- 10795278 TI - Caring in Alzheimer's disease. PMID- 10795280 TI - Towards a society for all ages. PMID- 10795279 TI - Role of operation theatre staff in prevention of infection. PMID- 10795281 TI - The United Nations and ageing. PMID- 10795282 TI - World Health Day theme. PMID- 10795283 TI - Human rights in the global context. PMID- 10795284 TI - Care provided by 'skill mix' and 'informal care givers' to critically ill patients. PMID- 10795285 TI - Television. The third parent: good or bad for children? PMID- 10795286 TI - Excellence in nursing education. Concepts, strategies and practices. PMID- 10795287 TI - Undergraduate nursing evaluation. The OSCE approach. PMID- 10795288 TI - Prevention of nosocomial infection. The role of nurse. PMID- 10795289 TI - Florence Nightingale. Pioneer of nursing profession. PMID- 10795290 TI - Palliative care. Devising new approaches to pain relief. PMID- 10795291 TI - Care of relatives during a patient's stay in the I.C.U. PMID- 10795292 TI - Learning from the developed world. An experience of nursing in Australia. PMID- 10795293 TI - Safe motherhood. Nurses' role in maternal and child health care. PMID- 10795294 TI - Ethics in nursing research. PMID- 10795296 TI - Hepatitis B. An occupational hazard. PMID- 10795295 TI - A to Z of backache. PMID- 10795297 TI - When your heart wants to remember: person centred dementia care. AB - The traditional approach to dementia care was essentially based around the view that so long as people with dementia were kept 'fed', clean and dry, that was all they needed. This minimalistic approach to care arose, not because of any intention to harm or to deprive people from care, but because of established knowledge, values and beliefs about dementia that have developed in UK society over the past 100 years (Harding and Palfrey 1997). This learning unit aims to enable nurses to develop a better understanding of person centred dementia care so that they might reflect more critically on their working with, and promote wellbeing for, people with dementia. It is necessary to watch the entire video before starting work on this Nursing Update workbook. PMID- 10795298 TI - Nice little earner. PMID- 10795299 TI - Will long division add up? PMID- 10795300 TI - A home for life. PMID- 10795301 TI - Back to basics wound care. PMID- 10795302 TI - Less pain, more gain. PMID- 10795303 TI - Upwardly mobile. PMID- 10795304 TI - Male catheterisation: 1. PMID- 10795305 TI - My working life. Independent living. PMID- 10795306 TI - Cloning of cDNA encoding NtEPc, a marker protein for the embryogenic dedifferentiation of immature tobacco pollen grains cultured in vitro. AB - We partially purified three Nicotiana tabacum L. embryogenic pollen-abundant phosphoproteins (NtEPa to c) which appeared in the cells undergoing a dedifferentiation process from immature pollen grains to embryogenic cells, caused by glutamine-deficiency in vitro. All the NtEPs had a highly conserved N terminal amino acid sequence. Using degenerate oligonucleotide probes designed from the amino acid sequences, the cDNA for NtEPc was isolated from a cDNA library of pollen cultured in glutamine-free medium The cDNA sequence showed moderate homology with several type-1 copper-binding glycoproteins and with a kind of early nodulin though its function could not be predicted. Expression analysis revealed that the level of mRNA for NtEPc was high during the dedifferentiation and also in the very early period of pollen embryogenesis but it was low in the developmental process of microspores/pollen in anthers, in the in vitro maturation process and both in the stational and logarithmic growth phases of tobacco BY-2 cells. Furthermore, an acidic medium pH, which promoted the induction of dedifferentiation increased the level of mRNA for NtEPc, whereas the presence of 6-benzylaminopurine, which inhibited it, decreased the level. These results suggest that the expression of NtEPc gene is correlated with the dedifferentiation but not with pollen development or cell division. PMID- 10795307 TI - Cooperation of photosystems II and I in leaves as analyzed by simultaneous measurements of chlorophyll fluorescence and transmittance at 800 nm. AB - Parallel measurements of CO2 assimilation, Chl fluorescence and 800 nm transmittance were carried out on intact leaves of wild type and cytochrome b6/f deficient transgenic tobacco grown at two different light intensities and temperatures, with the aim to diagnose processes limiting quantum yield of photosynthesis and investigate their adaptations to growth conditions. Relative optical cross-sections of PSII and PSI antennae were calculated from measured gas exchange rates and fluorescence-related losses at PSII and P700 oxidation-related losses at PSI. In nonstress conditions (high light grown wild type and low light grown antisense type) optimal relative optical cross-section of PSII (aII) was 0.48-0.51 and that of PSI (aI) was 0.38-0.40, leaving a non-photosynthetic absorption cross-section (a0) of 0.09-0.14 for nitrite assimilation and absorption in PSII beta and other photosynthetically inactive pigments. Stress conditions (low light grown wild type and high light grown antisense type, elevated growth temperatures) tend to increase a0 and decrease PSII antenna cross section more than that of PSI antenna, but this rule is reversed during senescence. PMID- 10795309 TI - Characterization of a zebra mutant of rice with increased susceptibility to light stress. AB - The rice zebra mutant TCM248 is a single recessive mutant. This mutant develops transverse-striped leaves with green and white sectors under alternate light/dark growth conditions. Mutants that were grown under a higher light intensity during the light period showed a more intense striped phenotype. The white tissues contained abnormal chloroplasts with few internal membrane structures, while the green tissues in the mutants contained normal chloroplasts. The white tissue contained only trace amounts of Chls and carotenoids, and mRNA accumulation of nuclear genes encoding chloroplast proteins (rbcS, cab) was strongly suppressed compared to that in the wild type plants. A series of growth condition shift experiments demonstrated that the mutant displayed the striped phenotype only if it was exposed to the alternate light/dark growth conditions during a limited stage of early leaf development. These data suggest that the zebra gene is involved in the acquisition of photoprotective capacity of the plants and that this gene functions at an early stage of chloroplast differentiation. PMID- 10795308 TI - Cytokinin-binding proteins from tobacco callus share homology with osmotin-like protein and an endochitinase. AB - To study the signal transduction of cytokinins, we characterized cytokinin binding proteins (CBPs) isolated from tobacco callus Nicotiana tabacum. Two high affinity CBPs, CBP1 and CBP2, were isolated from the soluble fraction of tobacco callus BY-2 cells by anion exchange chromatography on a DEAE-cellulose column and affinity chromatography on a benzyladenine (BA)-linked Sepharose 4B column. Cytokinin-binding activity was determined by the equilibrium dialysis method. The degree of purification of CBP1 and CBP2 was 270 and 600-fold, respectively. These proteins had molecular masses of 34 kDa and 26 kDa, and to bind benzyladenine (BA) with dissociation constants (Kd) of 8.9 x 10(-6) M and 1.1 x 10(-6) M, respectively. Binding of BA to CBP2 was inhibited by zeatin and kinetin but not by adenine, adenosine, ATP or IAA. The optimum pH for binding of BA to CBP1 and CBP2 was approximately pH 6.5 and 7.5, respectively. CBP1 showed significant homology (90%) with endochitinase and CBP2 with osmotin-like protein (OLP). These findings and the results of immunoblotting analysis and cytokinin-binding assay of recombinant OLP indicated that CBP2 is OLP, a stress protein. PMID- 10795310 TI - Wound-induced expression of a tobacco peroxidase is not enhanced by ethephon and suppressed by methyl jasmonate and coronatine. AB - In tobacco plants, wounding induces production of a set of defense-related proteins such as basic pathogenesis-related (PR) proteins and proteinase inhibitors (PIs) via the jasmonate/ethylene pathway. Although class III plant peroxidase (POX) is also wound-inducible, the regulatory mechanism for its wound induced expression is not fully understood. Here, we describe that a tobacco POX gene (tpoxN1), which is constitutively expressed in roots, is induced locally 30 min after wounding and then systemically in tobacco plants. Infection of necrotizing virus also induced tpoxN1 gene. The wound-induced expression was not enhanced by known wound-signal compounds such as methyl jasmonate (MeJA) and ethephon in contrast to other wound-inducible genes such as basic PR-1 and PI-II genes. And treatment with MeJA and coronatine, biological analogs of jasmonate, rather suppressed the tpoxN1 expression. Salicylic acid, an antagonist of jasmonate-based wound signaling, did not suppress the wound-induced expression of tpoxN1. Only spermine, which is reported as an endogenous inducer for acidic PR genes in tobacco mosaic virus-infected tobacco leaves, could induce tpoxN1 gene expression. These results suggest that wound-induced expression of the tpoxN1 gene is regulated differently from that of the basic PR and PI-II genes. PMID- 10795311 TI - Reversal of blue light-stimulated stomatal opening by green light. AB - Blue light-stimulated stomatal opening in detached epidermis of Vicia faba is reversed by green light. A 30 s green light pulse eliminated the transient opening stimulated by an immediately preceding blue light pulse. Opening was restored by a subsequent blue light pulse. An initial green light pulse did not alter the response to a subsequent blue light pulse. Reversal also occurred under continuous illumination, with or without a saturating red light background. The magnitude of the green light reversal depended on fluence rate, with full reversal observed at a green light fluence rate twice that of the blue light. Continuous green light given alone stimulated a slight stomatal opening, and had no effect on red light-stimulated opening. An action spectrum for the green light effect showed a maximum at 540 nm and minor peaks at 490 and 580 nm. This spectrum is similar to the action spectrum for blue light-stimulated stomatal opening, red-shifted by about 90 nm. The carotenoid zeaxanthin has been implicated as a photoreceptor for the stomatal blue light response. Blue/green reversibility might be explained by a pair of interconvertible zeaxanthin isomers, one absorbing in the blue and the other in the green, with the green absorbing form being the physiologically active one. PMID- 10795312 TI - Two novel intrinsic annexins accumulate in wheat membranes in response to low temperature. AB - Four immunologically related proteins that belong to the annexin family were identified in cold acclimated wheat (Triticum aestivum). Two soluble forms with molecular masses of 34 and 36 kDa were found to bind phospholipid membranes in a calcium-dependent manner. These two forms are similar to the previously reported doublet in several plant species. The other two forms, with molecular masses of 39 and 22.5 kDa, were found associated with the microsomal fraction. Biochemical analysis showed that both forms are intrinsic membrane proteins and their association with the membrane is calcium independent. This is, to our knowledge, the first report of the presence of these annexin forms in plants. Membrane purification by two phase partitioning demonstrated that the p39 form is localized to the plasma membrane. Immunoblot analysis showed that the protein level of both p39 and p22.5 increases gradually reaching a maximum level after one day of low temperature exposure. The protein accumulation was similar in both hardy and less hardy cultivars, suggesting that the accumulation is not correlated with freezing tolerance. The results are discussed with respect to the possible role of these new intrinsic membrane annexins in low temperature signal transduction pathway. PMID- 10795313 TI - A cysteine protease from maize isolated in a complex with cystatin. AB - We recently purified a latent but SDS-activated protease complex (40, 15- or 13 kDa proteins) from maize [Yamada et al. (1998) Plant Cell Physiol. 39: 106]. Here, we revealed that the complex was composed of a cysteine protease (40 kDa) and a cystatin, cysteine protease inhibitor (15- or 13-kDa). This is the first report on the isolation of a complex consisting of a cystatin and a target cysteine protease from plants. Cloning of the cysteine protease revealed that it had low homology (25-30%) to other maize cysteine proteases cloned to date but was highly homologous to other plant cysteine proteases such as rice oryzain alpha (84%) and the homologs (50-80%). The cysteine protease expressed in Escherichia coli showed the same substrate and inhibitor specificities as the protease of the complex, demonstrating that the isolated cDNA clone exactly encodes the protease of the complex. The protease expressed in E. coli itself was active but not latent, probably because it was not bound to cystatin. It is most likely that in vitro activation of the protease complex by SDS is caused by the release of bound cystatin. The mRNA of protease was expressed in various tissues except for seeds. PMID- 10795314 TI - Overexpression, enzymatic properties and tissue localization of a ferrochelatase of cucumber. AB - Ferrochelatase catalyzes the insertion of Fe2+ into protoporphyrin IX to generate protoheme. A putative mature region of a cucumber ferrochelatase cDNA (hemH) was overexpressed in Escherichia coli and purified to homogeneity (40 kDa). The optimum pH was 7.7, and the apparent K(m) values for deuteroporphyrin IX and Fe2+ were 14.4 microM and 4.7 microM, respectively. The activity of the ferrochelatase was inhibited by N-methylprotoporphyrin IX (I50 = 4 nM). Western blot analysis with a polyclonal antibody raised against the recombinant ferrochelatase showed that the antibody crossreacted with protein extracts from hypocotyls and roots of cucumber but not with that from cotyledons. The antibody did not crossreact with proteins of thylakoid membranes of chloroplasts in cucumber cotyledons, although the ferrochelatase activity was mainly associated with the thylakoid membranes. Northern blot analysis also indicated that the hemH gene was expressed mainly in hypocotyls and roots, but little in cotyledons, and the level of the hemH transcripts was not light-responsive. These results demonstrated that the cucumber hemH gene encodes a ferrochelatase which presumably functions for heme biosynthesis in non-photosynthetic tissues, such as hypocotyls and roots, and suggested the presence of other types of ferrochelatase in cucumber, one of which is located in thylakoid membranes of chloroplasts. PMID- 10795315 TI - Purification and characterization of beta-cyanoalanine synthase and cysteine synthases from potato tubers: are beta-cyanoalanine synthase and mitochondrial cysteine synthase same enzyme? AB - beta-Cyanoalanine synthase (CAS; EC 4.4.1.9) and two kinds of cysteine synthases (CS; EC 4.2.99.8) have been purified from the particulate fraction of potato tubers. By DEAE Sephacel and Resource PHE chromatography, CAS activity was separated from two CS activities, designated as CS-1 and CS-2. The molecular masses of CAS, CS-1 and CS-2 were estimated to be 37, 39 and 34 kDa, respectively, by SDS-PAGE analysis. The purified CAS had CS activity, and both CS 1 and CS-2 had CAS activity. However, CAS and CSs had significant differences in kinetic characters. The antibody raised against purified CAS discriminated CAS from CSs, whereas the antibody raised against purified CS-2 recognized CS-1 and CS-2 but not CAS. The molecular mass and the partial amino acid sequence of CS-2 were similar to those of the cytosolic CS of potato, whereas the molecular mass of CS-1 was similar to that of the plastidic CS. The partial amino acid sequence of CAS was similar to those of CS isozymes, especially the mitochondrial CS isolated from spinach. PMID- 10795316 TI - Kinetic analysis of the K(+)-selective outward rectifier in Arabidopsis mesophyll cells: a comparison with other plant species. AB - This paper gives a kinetic analysis of the K(+)-selective outward-rectifier (IK,out) in the plasma membrane of Arabidopsis thaliana mesophyll cells in terms of the Hodgkin-Huxley formalism. We compared the kinetic characteristics of IK,out in Arabidopsis with IK,out channels in three other plant species that were subjected to a similar analysis: tobacco suspension cells, Vicia faba guard cells and Plantago media root cells. Because the activation kinetics of IK,out shows a clear voltage dependence, the time constant of half-activation (tau 1/2) and the elementary rate constant of channel opening (a) were calculated at the potential of half-activation (V1/2). The Arabidopsis IK,out activates relatively slowly and this is reflected in a tau 1/2 of approximately 1 s. The reason for this slow activation is twofold. Firstly, the value of a is 1.5 s-1 falls at the lower end of the range of values obtained for tobacco, Vicia and Plantago: 1.1 to 3.0 s-1. Secondly, IK,out in Arabidopsis has four closed states, while tobacco and Vicia have only two. As observed in other plant species, the activation kinetics of IK,out in Arabidopsis are sensitive to external K+: V1/2 shifts with EK but remains approximately 50 mV more positive than EK. PMID- 10795317 TI - Tissue and intracellular localization of indican and the purification and characterization of indican synthase from indigo plants. AB - Indican (indoxyl beta-D-glucoside) was found to accumulate only in green leaves of the indigo plant, and not in any other tissues. Comparisons of the indican content of protoplasts and vacuoles showed that indican was stored only in the vacuole of the cell. Indican content appeared and increased with the appearance and growth of leaves. In mature plants, the younger leaves contained larger amounts of indican than the older ones. Cell extracts of young leaves of indigo plant catalyzed the synthesis of indican from UDP-glucose and indoxyl. Indican synthase was extracted and purified from young leaves. The enzyme was separated into two fractions by anion-exchange chromatography. The enzyme in the fraction which was eluted by 0.1 M NaCl had a molecular weight of 53,000 by SDS-PAGE. Optimum pH of the enzyme was at about 10.0, indicating that the enzyme is likely localized in a different intracellular compartment from that of indican storage. The enzyme showed normal Michaelis-Menten kinetics and a K(m) value of 0.13 mM for UDP-glucose. PMID- 10795318 TI - Expression of NADPH-Protochlorophyllide oxidoreductase gene in fully green leaves of cucumber. AB - Photoreduction of protochlorophyllide a to chlorophyllide a that is catalyzed by NADPH-protochlorophyllide oxidoreductase (NPR) is only a light-dependent step in overall processes of biosynthesis of Chl in angiosperms. Unlike many other plants, in fully green leaves of cucumber the expression of only a single NPR gene of cucumber was positively regulated by light and developmental age of leaf, and also by diurnal and circadian rhythms. The single NPR gene was shown to be involved in Chl synthesis throughout of the vegetative growth of cucumber (in cotyledons and fully green leaves). PMID- 10795319 TI - A pulse of blue light induces a transient increase in activity of apoplastic K+ in laminar pulvinus of Phaseolus vulgaris L. AB - A pulse of blue light induced both a transient increase in activity of apoplastic K+ and membrane depolarization in laminar pulvinus of Phaseolus vulgaris L. This shows that blue-light-induced net efflux of K+ from motor cells is closely related to membrane depolarization. PMID- 10795320 TI - Cytochalasin A inhibits the binding of phenylalanine ammonia-lyase mRNA to ribosomes during induction of phytoalexin in pea seedlings. AB - Cytochalasin A (CA) blocked the accumulation of phytoalexin and phenylalanine ammonia-lyase (PAL)-protein in pea tissues treated with a fungal elicitor but scarcely affected the PAL-mRNA content. Further analysis showed that CA decreased the PAL-mRNA bound to ribosomes. These results indicate that actin filaments are tightly associated with the translational process of the PAL gene. PMID- 10795321 TI - Analysis of a mod B mutant in Dictyostelium discoideum using mRNA differential display. AB - Three differential cDNAs of Dictyostelium, not detected in the mod B mutant defective in O-glycosylation, were isolated by using an mRNA differential display. These cDNAs encode a protein tyrosine kinase, an adenylyl cyclase and a putative protein kinase C inhibitor whose expression is developmentally regulated. PMID- 10795322 TI - Time-sequence observations of microtubule dynamics throughout mitosis in living cell suspensions of stable transgenic Arabidopsis--direct evidence for the origin of cortical microtubules at M/G1 interface. AB - Transgenic Arabidopsis thaliana, stably expressing a GFP-TUA6 fusion protein, were subcultured in B5 medium supplemented with 2,4-D and BA. In the cell suspensions, the microtubular changes in the mitotic cells could be monitored by time-sequence observations using a time-lapse system of fluorescence microscopy. We have succeeded in following the microtubule (MT) dynamics in living cells throughout mitosis, from the late G2 phase to early G1 phase, and found that, at the M/G1 interface, the cortical MTs were firstly reorganized in the perinuclear regions and then in the cortex, as we had previously suggested (Hasezawa and Nagata 1991, Nagata et al. 1994). The significance of this observation on the origin of cortical MTs is discussed. PMID- 10795323 TI - [Is there a better treatment for hypothyroidism?]. PMID- 10795324 TI - [Primary parvovirus B19 infection in immunocompetent adults: clinical and biological manifestations. Retrospective study of 16 patients]. AB - PURPOSE: The spectrum of clinical and biological expressions of parvovirus B19 infection is incompletely described. METHODS: We report 16 retrospective cases of primary parvovirus B19 infection that occurred in immunocompetent adults and were diagnosed by serology. RESULTS AND CONCLUSION: Most cases conformed to the classical descriptions of parvovirus-induced arthropathy (transient polyarthritis often accompanied by erythema, dysesthesia and pruritus). However, the diagnosis of primary parvovirus B19 infection should also be considered in other clinical and biological situations such as edema, acute anemia or cytopenia, vascular purpura, neuro-ophthalmologic manifestations, and hepatitis. Parvovirus infection may mimic diseases such as lupus erythematosus or systemic necrotizing vasculitis. The risk for confusion with both diseases is increased, as immunological abnormalities such as hypocomplementemia, when they are systematically searched for, prove to be common. PMID- 10795325 TI - [Mediastinal fibrosis combined with Behcet's disease. Three case reports]. AB - PURPOSE: Intrathoracic manifestations other then vascular aneurysm and thrombosis are rare in the course of Behcet's disease. Mediastinal fibrosis is also unusual. METHODS: Three cases of Behcet's disease with initial mediastinal fibrosis are reported. Diagnosis means, pathogenic mechanisms and therapeutic modalities are also more precisely defined. RESULTS: Three male patients (21, 25 and 27 years of age, respectively) were hospitalized for progressive vena cava superior syndrome accompanied by mediastinal fibrosis demonstrated by MRI and/or CT scan which both showed infiltration of the mediastinal fat. Multiple calcifications were also present in one patient. Histology was performed in one case. Thrombosis of the vena cava superior and innominata veins was associated with fibrosis in all three cases. Thrombosis of venae subclavia and left jugularis was also present in one case. Other manifestations leading to the diagnosis of Behcet's disease were demonstrated by the patient's history and, in one case, occurred on the fourth day of hospitalization. After initiation of a treatment including corticoids, anticoagulants and colchicine, the outcome was favorable in two cases. The third patient died. CONCLUSION: Mediastinal fibrosis combined with Behcet's disease has rarely been described. However, it should be included in manifestations of the disease. The causative process which might be vasculitis has to be demonstrated. PMID- 10795327 TI - [Methicillin-resistant Staphylococcus aureus: factors responsible for its incidence]. AB - INTRODUCTION: How can we explain that the proportion of methicillin-resistant Staphylococcus aureus (MRSA) varies between the European countries, ranging from < 1% in Scandinavia to > 30% in Spain, France and Italy? This paper is aimed at attempting to determine factors at the origin of the spreading of endemic MRSA strains as of the early 1980s. Those strains are characterized by their ability to develop resistance to current antibiotics and make treatment of severe and deep infections more complex. CURRENT KNOWLEDGE AND KEY POINTS: Differences in the virulence of MRSA strains and that of susceptible strains appear unlikely. MRSA prevalence seems to be a growing problem, especially in Southern Europe where rates of resistance to other anti-staphylococcal antibiotics are high. General policies for antibiotic therapy as well as the implementation of strategies for prevention and control of MRSA might be responsible for such rates. Indeed, once MRSA is introduced into a facility without control program, this multiresistant bacteria rapidly spreads within the hospital and becomes endemic, expanding its reservoir. FUTURE PROSPECTS ET PROJECTS: Due to the introduction of new methods in microbiology and communication, infection control measures including procedures for isolation and identification of MRSA reservoirs are still feasible; however, their implementation requires human and material resources. Areas requiring improvement in the detection of MRSA outbreaks are identified in this paper, with particular emphasis on the need for national surveillance of MRSA prevalence and reappraisal of MRSA control strategies in French hospitals. PMID- 10795326 TI - [Retrospective multivariate analysis of radio-pathological correlations of nonpalpable breast lesions. Experience of the Hospital Saint-Louis]. AB - PURPOSE: Several studies have demonstrated that systematic breast cancer screening increases overall survival. We report our experience regarding diagnosis of breast lesions detected using mammography. METHODS: Case reports of patients operated on in either 1992 or 1993 were retrospectively reviewed. A multivariate analysis of the clinico-pathological correlation was performed. RESULTS: Four hundred fifty seven patients representing on total 544 procedures, were included in the study. Mean age was 50.5 years (range 19-80 years). Most of the patients had no previous history of mammary lesion. Mammography was performed with prophylactic intent in more than 60% of the cases. Four hundred twelve (75.7%) benign lesions were diagnosed. Main lesions were: adenofibroma (15.7%), fibrocystic mastopathy (66.3%), adenosis (26.2%), ductal hyperplasia (23.9%), lobular hyperplasia (10.7%), and combined ductal and lobular hyperplasia (8.5%). Hyperplasia accompanied by cytonuclear atypia was observed in 49 (11%) cases. One hundred thirty two (24.3%) malignant lesions were reported, including 69 (52.3%) invasive carcinomas and 63 (47.7%) in situ carcinomas. Only nine axillary lymph node dissections were positive and 75 minimal breast cancers were diagnosed. The multivariate analysis showed that only radiological signs are a risk factor for cancer. The relative risk for cancer when focus of irregular and vermicular microcalcifications are diagnosed is 4.2 (2.0-8.5). It is 5.6 (2.5-12.5) in case of spiculated opacity. CONCLUSION: Exeresis following radiological prophylactic screening allows diagnosis of high-risk benign lesions and low-stage breast cancer. Radiological parameters are the most powerful predictive factors for malignancy. PMID- 10795328 TI - [Vascular manifestations of Behcet's syndrome associated with solitary ulcerations and resolved with immunosuppressants]. AB - INTRODUCTION: Behcet's disease is a systemic inflammatory disorder characterized by vasculitis. Its typical features are recurrent oral and genital ulcerations with uveitis. Although vascular lesions are not listed among the criteria for diagnosis of Behcet's disease, up to 25-35% of the patients develop complications in arterial and venous large vessels. EXEGESIS: We describe the case of a 45-year old French man with Behcet-type vasculopathy. Though only one sign of Behcet's disease, i.e., oral ulcerations, was present, the patient had to undergo emergency surgery three times. The postoperative treatment combined corticosteroids, azathioprine, and oral anticoagulants. Three years later no therapy failure was observed. CONCLUSION: On the basis of 1) recurrent aortic aneurysms, 2) large arterial and venous occlusive lesions, 3) superficial phlebitis, and 4) ulcerations of the aorta in macro- and microscopic examination of resected aortic walls, we concluded that the patient's life threatening vasculopathy was of the Behcet's type, even if several of the diagnostic features of Behcet's disease were lacking. Variations in clinical features of Behcet's disease are observed that might be due to hereditary traits, particularly to the genetic expression of an incomplete phenotype that would lead to the lack of typical clinical features. PMID- 10795329 TI - [A case of hepatic Loeffler's syndrome]. AB - INTRODUCTION: During nematode infection transit of the microorganism in the lung may be associated with transient infiltration lesions accompanied by marked eosinophilia. In the invasion stage of the disease maturation and sexual differentiation of schistosomules is restricted to portal vessels. Liver ultrasonography may show concomitant abnormalities. EXEGESIS: Eosinophilia was diagnosed in a 36-year-old man residing in Central Africa. Clinical examination, chest X-ray and biology were normal. Liver ultrasonography showed numerous hypoechoic lesions, up to 3 cm in diameter. Ultrasonography-directed biopsy demonstrated infiltration of both portal vessels and liver sinusoids, with sometimes microabcesses. No parasite was found in either blood, stool, or rectal biopsy. Immunoelectrophoresis and hemagglutination showed positive bilharziosis serology, whereas serology for other helminthiases was negative. Four months after treatment eosinophilia was still high, liver ultrasonography was normal, and hemagglutination was negative. CONCLUSION: The transient shistosomules maturation-related liver abnormalities detected by ultrasonography should be differentiated from the laying phase of the disease. PMID- 10795330 TI - [Medical responsibility and nosocomial infections]. AB - Modifications of the civil and administrative jurisprudence regarding nosocomial infections reflect the will to pay the patient for compensation for injury. The June 29, 1999 decree of the French Court of Cessation shows the transition from presumption of malpractice to obligation of safety. The authors describe those modifications, including the physician's and hospital liabilities, and assess their consequences, as proof of an external cause is now mandatory while proof of absence of malpractice is no longer sufficient. PMID- 10795331 TI - [Diagnosis of infiltrating breast cancer]. PMID- 10795332 TI - [A swollen knee]. PMID- 10795333 TI - [Thrombotic manifestations revealing a digestive tract disease]. PMID- 10795334 TI - Factors in risk perception AB - Risk perception is a phenomenon in search of an explanation. Several approaches are discussed in this paper. Technical risk estimates are sometimes a potent factor in accounting for perceived risk, but in many important applications it is not. Heuristics and biases, mainly availability, account for only a minor portion of risk perception, and media contents have not been clearly implicated in risk perception. The psychometric model is probably the leading contender in the field, but its explanatory value is only around 20% of the variance of raw data. Adding a factor of "unnatural risk" considerably improves the psychometric model. Cultural Theory, on the other hand, has not been able to explain more than 5-10% of the variance of perceived risk, and other value scales have similarly failed. A model is proposed in which attitude, risk sensitivity, and specific fear are used as explanatory variables; this model seems to explain well over 30-40% of the variance and is thus more promising than previous approaches. The model offers a different type of psychological explanation of risk perception, and it has many implications, e.g., a different approach to the relationship between attitude and perceived risk, as compared with the usual cognitive analysis of attitude. PMID- 10795336 TI - Cultural myths of human and physical nature: integrated or separated? AB - Cultural Theory distinguishes between myths of human and physical nature as two integrated aspects of four cultural biases: hierarchy, egalitarianism, individualism, and fatalism. These biases serve as individuals' key orientations toward, for example, risk perception, public policy, and political preference. Myths of human and physical nature draw upon different intellectual histories, and an epistemological merger between the two aspects is not unproblematic. A self-administered mail survey of organized environmentalists in Norway included the theory's graphical description of myths of physical nature and verbal descriptions of myths of human nature. The respondents understood the logic of the myths of physical nature well and did not have problems in ranking them, thereby disconfirming the theory's claim that any of the myths appear irrational from the perspective of any other. The empirical results show that respondents gave the highest priority to the hierarchical myth of physical nature and that they also endorsed the egalitarian myth of human nature. Although this outcome may appear irrational from a theoretical perspective, the authors argue that (Grid/group) Cultural Theory is best served by treating the myths of physical and human nature as logically independent of one another. PMID- 10795335 TI - Correlates of household seismic hazard adjustment adoption. AB - This study examined the relationships of self-reported adoption of 12 seismic hazard adjustments (pre-impact actions to reduce danger to persons and property) with respondents' demographic characteristics, perceived risk, perceived hazard knowledge, perceived protection responsibility, and perceived attributes of the hazard adjustments. Consistent with theoretical predictions, perceived attributes of the hazard adjustments differentiated among the adjustments and had stronger correlations with adoption than any of the other predictors. These results identify the adjustments and attributes that emergency managers should address to have the greatest impact on improving household adjustment to earthquake hazard. PMID- 10795337 TI - Development of a single-meal fish consumption advisory for methyl mercury. AB - Methyl mercury (meHg) contamination of fish is the leading cause of fish consumption advisories in the United States. These advisories have focused upon repeated or chronic exposure, whereas risks during pregnancy may also exist from a single-meal exposure if the fish tissue concentration is high enough. In this study, acute exposure to meHg from a single fish meal was analyzed by using the one-compartment meHg biokinetic model to predict maternal hair concentrations. These concentrations were evaluated against the mercury hair concentration corresponding to the U.S. Environmental Protection Agency's reference dose (RfD), which is intended to protect against neurodevelopmental effects. The one compartment model was validated against blood concentrations from three datasets in which human subjects ingested meHg in fish, either as a single meal or multiple meals. Model simulations of the single-meal scenario at different fish meHg concentrations found that concentrations of 2.0 ppm or higher can be associated with maternal hair concentrations elevated above the RfD level for days to weeks during gestation. A single-meal fish concentration cutoff of > or = 2.0 ppm is an important consideration, especially because this single high exposure event might be in addition to a baseline meHg body burden from other types of fish consumption. This type of single-meal advisory requires that fish sampling programs provide data for individual rather than composited fish, and take into account seasonal differences that may exist in fish concentrations. PMID- 10795338 TI - Categorizing risks for risk ranking. AB - Any practical process of risk ranking must group hazards into a manageable number of categories. Defining such categories requires value choices that can have important implications for the rankings that result. Most risk-management organizations will find it useful to begin defining categories in terms of environmental loadings or initiating events. However, the resulting categories typically need to be modified in light of other considerations. Risk-ranking projects can benefit from considering several alternative categorization strategies and drawing upon elements of each in developing their final categorization of risks. In principle, conducting multiple ranking exercises by using different categorizations could be interesting and useful. In practice, agencies are unlikely to have either the resources or patience to do this, but other groups in society might. Done well, such additional independent rankings could add valuable inputs to democratic risk-management decision making. PMID- 10795339 TI - Random number generation from right-skewed, symmetric, and left-skewed distributions. AB - Monte Carlo simulations have become a mainstream technique for environmental and technical risk assessments. Because their results are dependent on the quality of the involved input distributions, it is important to identify distributions that are flexible enough to model all relevant data yet efficient enough to allow thousands of evaluations necessary in a typical simulation analysis. It has been shown in recent years that the S-distribution provides accurate representations for frequency data that are symmetric or skewed to either side. This flexibility makes the S-distribution an ideal candidate for Monte Carlo analyses. To use the distribution effectively, methods must be available for drawing S-distributed random numbers. Such a method is proposed here. It is shown that S-distributed random numbers can be efficiently generated from a simple algebraic formula whose coefficients are tabulated. The method is shown step by step and illustrated with a detailed example. (The tables are accessible in electronic form in the FTP parent directory at http:@www.musc.edu/voiteo/ftp/.) PMID- 10795340 TI - Risk analysis of PCB exposure via the soil-food crop pathway, and alternatives for remediation at Serpukhov, Russian Federation. AB - A risk assessment was conducted to determine the likelihood of certain health risks resulting from exposure to soils and food crops contaminated with polychlorinated biphenyl (PCBs). PCBs have contaminated soils, river sediments, and air in the past as a result of industrial activities at a capacitor plant located in the City of Serpukhov, Russian Federation. This risk assessment and suggestions for remediation are designed to aid in decision-making efforts by a joint Russian-American research team developing a community, national, and international response to industrial contamination. Bobovnikova et al. (The Science of the Total Environment 139/140, 357-364, [1993]) have reported that PCBs are elevated in soils and sediments, breast milk, and locally grown foods in the Serpukhov area. Data from these and other investigators have been used in this risk assessment to calculate a potential cancer risk resulting from exposure to PCBs. Our assessment indicates that members of the local population may be at increased risk of cancer, and possibly other adverse health effects, as a result of PCB contamination of their environment. Because previously unassessed environmental contamination is a common problem in the former Soviet Republics, as well as many other areas of the world, we believe this type of evaluation, using known methods, can serve as a model for assessment efforts in other parts of the globe and result in remediative efforts in regions constrained by faltering economies. PMID- 10795341 TI - Cancer risk estimation for mixtures of coal tars and benzo(a)pyrene. AB - Two-year chronic bioassays were conducted by using B6C3F1 female mice fed several concentrations of two different mixtures of coal tars from manufactured gas waste sites or benzo(a)pyrene (BaP). The purpose of the study was to obtain estimates of cancer potency of coal tar mixtures, by using conventional regulatory methods, for use in manufactured gas waste site remediation. A secondary purpose was to investigate the validity of using the concentration of a single potent carcinogen, in this case benzo(a)pyrene, to estimate the relative risk for a coal tar mixture. The study has shown that BaP dominates the cancer risk when its concentration is greater than 6,300 ppm in the coal tar mixture. In this case the most sensitive tissue site is the forestomach. Using low-dose linear extrapolation, the lifetime cancer risk for humans is estimated to be: Risk < 1.03 x 10(-4) (ppm coal tar in total diet) + 240 x 10(-4) (ppm BaP in total diet), based on forestomach tumors. If the BaP concentration in the coal tar mixture is less than 6,300 ppm, the more likely case, then lung tumors provide the largest estimated upper limit of risk, Risk < 2.55 x 10(-4) (ppm coal tar in total diet), with no contribution of BaP to lung tumors. The upper limit of the cancer potency (slope factor) for lifetime oral exposure to benzo(a)pyrene is 1.2 x 10(-3) per microgram per kg body weight per day from this Good Laboratory Practice (GLP) study compared with the current value of 7.3 x 10(-3) per microgram per kg body weight per day listed in the U.S. EPA Integrated Risk Information System. PMID- 10795342 TI - Risk estimation and value-of-information analysis for three proposed genetic screening programs for chronic beryllium disease prevention. AB - Genetic differences (polymorphisms) among members of a population are thought to influence susceptibility to various environmental exposures. In practice, however, this information is rarely incorporated into quantitative risk assessment and risk management. We describe an analytic framework for predicting the risk reduction and value-of-information (VOI) resulting from specific risk management applications of genetic biomarkers, and we apply the framework to the example of occupational chronic beryllium disease (CBD), an immune-mediated pulmonary granulomatous disease. One described Human Leukocyte Antigen gene variant, HLA-DP beta 1*0201, contains a substitution of glutamate for lysine at position 69 that appears to have high sensitivity (approximately 94%) but low specificity (approximately 70%) with respect to CBD among individuals occupationally exposed to respirable beryllium. The expected postintervention CBD prevalence rates for using the genetic variant (1) as a required job placement screen, (2) as a medical screen for semiannual in place of annual lymphocyte proliferation testing, or (3) as a voluntary job placement screen are 0.08%, 0.8%, and 0.6%, respectively, in a hypothetical cohort with 1% baseline CBD prevalence. VOI analysis is used to examine the reduction in total social cost, calculated as the net value of disease reduction and financial expenditures, expected for proposed CBD intervention programs based on the genetic susceptibility test. For the example cohort, the expected net VOI per beryllium worker for genetically based testing and intervention is $13,000, $1,800, and $5,100, respectively, based on a health valuation of $1.45 million per CBD case avoided. VOI results for alternative CBD evaluations are also presented. Despite large parameter uncertainty, probabilistic analysis predicts generally positive utility for each of the three evaluated programs when avoidance of a CBD case is valued at $1 million or higher. Although the utility of a proposed risk management program may be evaluated solely in terms of risk reduction and financial costs, decisions about genetic testing and program implementation must also consider serious social, legal, and ethical factors. PMID- 10795344 TI - Risk of extreme events in multiobjective decision trees. Part 1. Severe events AB - Earlier work with decision trees identified nonseparability as an obstacle to minimizing the conditional expected value, a measure of the risk of extreme events, by the well-known method of averaging out and folding back. This first of two companion papers addresses the conditional expected value that is defined as the expected outcome assuming the exceedance of a threshold beta, where beta is preselected by the decision maker. An approach is proposed to overcome the need to evaluate all policies in order to identify the optimal policy. The approach is based on the insight that the conditional expected value is separable into two constituent elements of risk and can thus be optimized along with other objectives, including the unconditional expected value of the outcome, by using a multiobjective decision tree. An example of sequential decision making for improving highway capacity is given. PMID- 10795343 TI - The assessment of radiation exposures in Native American communities from nuclear weapons testing in Nevada. AB - Native Americans residing in a broad region downwind from the Nevada Test Site during the 1950s and 1960s received significant radiation exposures from nuclear weapons testing. Because of differences in diet, activities, and housing, their radiation exposures are only very imperfectly represented in the Department of Energy dose reconstructions. There are important missing pathways, including exposures to radioactive iodine from eating small game. The dose reconstruction model assumptions about cattle feeding practices across a year are unlikely to apply to the native communities as are other model assumptions about diet. Thus exposures from drinking milk and eating vegetables have not yet been properly estimated for these communities. Through consultations with members of the affected communities, these deficiencies could be corrected and the dose reconstruction extended to Native Americans. An illustration of the feasibility of extending the dose reconstruction is provided by a sample calculation to estimate radiation exposures to the thyroid from eating radio-iodine-contaminated rabbit thyroids after the Sedan test. The illustration is continued with a discussion of how the calculation results may be used to make estimates for other tests and other locations. PMID- 10795345 TI - Risk of extreme events in multiobjective decision trees. Part 2. Rare events AB - Earlier work with decision trees identified nonseparability as an obstacle to minimizing the conditional expected value, a measure of the risk of extreme events, by the well-known method of averaging out and folding back. This second of two companion papers addresses the conditional expected value that is defined as the expected outcome assuming that a random variable is observed only in the upper 100 (1 - alpha) percent of potential outcomes, where alpha is a cumulative probability preselected by the decision maker. An approach is proposed to overcome the need to evaluate all policies in order to identify the optimal policy. The approach is based in part on approximating the conditional expected value by using statistics of extremes. An existing convenient approximation of the conditional expected value is shown to be separable into two constituent elements of risk and can thus be optimized, along with other objectives including the unconditional expected value of the outcome, in a multiobjective decision tree. An example of sequential decision making for remediation or environmental contamination is provided. The importance of the results for risk analyis beyond the minimization of conditional expected values is pointed out. PMID- 10795346 TI - Improving cancer dose-response characterization by using physiologically based pharmacokinetic modeling: an analysis of pooled data for acrylonitrile-induced brain tumors to assess cancer potency in the rat. AB - Historically, U.S. regulators have derived cancer slope factors by using applied dose and tumor response data from a single key bioassay or by averaging the cancer slope factors of several key bioassays. Recent changes in U.S. Environmental Protection Agency (EPA) guidelines for cancer risk assessment have acknowledged the value of better use of mechanistic data and better dose-response characterization. However, agency guidelines may benefit from additional considerations presented in this paper. An exploratory study was conducted by using rat brain tumor data for acrylonitrile (AN) to investigate the use of physiologically based pharmacokinetic (PBPK) modeling along with pooling of dose response data across routes of exposure as a means for improving carcinogen risk assessment methods. In this study, two contrasting assessments were conducted for AN-induced brain tumors in the rat on the basis of (1) the EPA's approach, the dose-response relationship was characterized by using administered dose/concentration for each of the key studies assessed individually; and (2) an analysis of the pooled data, the dose-response relationship was characterized by using PBPK-derived internal dose measures for a combined database of ten bioassays. The cancer potencies predicted for AN by the contrasting assessments are remarkably different (i.e., risk-specific doses differ by as much as two to four orders of magnitude), with the pooled data assessments yielding lower values. This result suggests that current carcinogen risk assessment practices overestimate AN cancer potency. This methodology should be equally applicable to other data-rich chemicals in identifying (1) a useful dose measure, (2) an appropriate dose-response model, (3) an acceptable point of departure, and (4) an appropriate method of extrapolation from the range of observation to the range of prediction when a chemical's mode of action remains uncertain. PMID- 10795347 TI - New British guidelines on the clinical management of drug misuse and drug dependence. PMID- 10795349 TI - Conversation with Joy Mott. PMID- 10795348 TI - Regulation of liquor licensing should be made to work. PMID- 10795350 TI - Beverage-specific alcohol consumption and cirrhosis mortality in a group of English-speaking beer-drinking countries. AB - AIMS: To compare beverage-specific per capita consumption and total alcohol consumption's associations with cirrhosis mortality rates in multiple countries. DESIGN: Pooled cross-sectional time-series analysis. SETTING: Australia, Canada, New Zealand, the United Kingdom and the United States during the years 1953-1993. MEASUREMENTS: National level data on per capita total alcohol, beer, wine and spirits consumption and standardized all-cause cirrhosis mortality rates. FINDINGS: Significant associations with cirrhosis mortality are found for both total ethanol and spirits. Spirits consumption is found to make up the majority of the effect of alcoholic beverage consumption on cirrhosis mortality and the model including only spirits is found to fit the data at least as well as the model including only total ethanol consumption. The lag relationship between all alcohol types and cirrhosis is found to be short with only present and 1 year's lagged consumption having significant associations. CONCLUSIONS: Spirits consumption rather than beer or wine is associated with cirrhosis mortality in this group of primarily beer-drinking countries. This finding offers important clues to understanding the drinking behaviors associated with cirrhosis mortality on the individual level. PMID- 10795351 TI - The epiphenomena of beverage-specific effects. PMID- 10795352 TI - Who fares poorly with alcoholic cirrhosis? PMID- 10795353 TI - Alcohol consumption, time-series methodology and disease outcomes. PMID- 10795354 TI - Explaining trends in liver cirrhosis. PMID- 10795356 TI - Over-serving patrons in licensed premises in Stockholm. AB - AIMS: To study the frequency of alcohol service to intoxicated patrons in licensed premises. DESIGN: Actors were hired to simulate severe intoxication. Following a review of videotaped portrayals of drunken behaviour by the actors, an expert panel of bartenders and restaurant managers determined which scene was least consistent with responsible beverage service. The actors then enacted this scene in the licensed premises, while monitored by project staff. SETTING: Ninety two licensed premises in Stockholm city were visited by the actors. FINDINGS: The actors were served alcohol at 95% of the licensed premises. CONCLUSION: The results show that existing serving policies at licensed premises in Stockholm are very permissive regarding the degree of intoxication among patrons. PMID- 10795357 TI - Fiesta as tradition, fiesta as change: ritual, alcohol and violence in a Mexican community. AB - AIMS: To explore the traditional Mexican religious fiesta as a primary locus for community-based excessive drinking and violence against women. The research argues that the pattern of drinking and violence cannot be placed within explanations that tie a breakdown in social norms to drinking nor drinking to a breakdown in social norms. DESIGN: The data were gathered over 14 continuous months of participant observation in 1995 and 1996, followed by two summer research projects in 1997 and 1998. The researcher documented the activities of the participants at each fiesta and followed-up each event with interviews of the parties involved in violent confrontations. In addition, life stories and archival data on violence were conducted and used to situate current findings against historical data. SETTING: All data were collected in the community of Santa Maria Atzompa. Atzompa is a cabecera or municipal seat, for six colonias (large neighborhoods), and three ranchos (agriculturally based communities) and has a population of just over 5200. The main community of Atzompa, where most of the research was conducted, has a population of 2700+. PARTICIPANTS: Over 1000 individuals participated in community religious fiestas that the researcher attended. More than half of these were men, and almost all the men present participated in the binge drinking. MEASUREMENTS: Sixteen religious fiestas and 13 non-religious fiestas were documented through participant observation and photographs. FINDINGS: In every religious-based fiesta, violence erupted and was directed primarily against women. Husband and wife abuse accounted for 10 of the 16 violent disruptions (63%) while male/male abuse accounted for six (38%). CONCLUSIONS: The perpetuation of binge drinking and violence are part of a historic cycle of male dominance that dates back to the introduction of alcohol distillation during colonization by the Spaniards in the 16th century, compounded today by frustration over their inability to control the economic and political aspects of their households and community. Women's acquiescence to the violence is a form of mediation over male frustrations that allow women to continue in their roles as economic providers. PMID- 10795358 TI - Quality of heroin and heroin-related deaths from 1987 to 1995 in Vienna, Austria. AB - AIMS: To investigate the quality of heroin seized in Vienna between 1987 and 1995 and to examine whether there was a relationship between the quality of heroin and the rate of drug-related deaths. DESIGN: Reports of heroin seizure analysis and post-mortem reports of heroin-related deaths in Vienna from 1987 to 1995 were analysed. RESULTS: There were 386 seizures of heroin comprising a total weight of 25 640.12 g heroin. All the seizures were in the base form. All seizures also contained a diluent, mainly lactose. Additionally, in 95 seizures caffeine, in four seizures paracetamol and in three seizures metaqualon were detected. Of a total of 764 drug-related deaths 506 cases were classified as heroin-related deaths. In 336 cases other central nervous system-depressant drugs and/or alcohol could be detected in the blood in addition to morphine (polydrug heroin-related deaths). There was evidence of a trend towards greater polydrug involvement during the study period. The age of victims of polydrug heroin-related deaths remained unchanged over time while the age of victims of pure heroin-related deaths decreased significantly. The morphine concentration in the medulla oblongata in heroin-related deaths decreased significantly. Finally, there was no statistically significant relationship between the rate of heroin-related deaths per year and the diacetylmorphine concentration of heroin seizures in that year. CONCLUSION: The results did not substantiate the widely held belief that increases in heroin-related deaths could be explained by an increase in the quality of heroin. PMID- 10795359 TI - Prevalence, incidence and risk factors of anaemia in HIV-positive and HIV negative drug users. AB - AIMS: To determine the prevalence of anaemia among HIV-positive and -negative drug users, estimate the incidence of anaemia among HIV-positive drug users and study risk factors for prevalent anaemia with emphasis on HIV infection, gender and drug use behaviour. DESIGN, SETTING AND PARTICIPANTS: Cross-sectional study among HIV-positive and HIV-negative drug users (n = 360) and a longitudinal study among HIV-positive drug users (n = 175, 860 visits) participating in a cohort study of drug users in Amsterdam. MEASUREMENTS: Self-reported data on socio demographics, drug use behaviour and zidovudine and protease inhibitor therapy on regular cohort visits. Laboratory determinations of haemoglobin concentration, CD4(+)-cell counts and HIV-1 RNA. FINDINGS: The prevalence of anaemia was 21.1% among participating drug users according to the WHO definition. The incidence of anaemia in HIV-positive individuals was 33.0/100 person-years. The main risk factor for anaemia was high frequency of injecting drugs. A Quetelet body mass index < or = 20 and female gender were also positively and significantly related to anaemia in the cross-sectional study. In the longitudinal study among HIV positive drug users progression of HIV infection indicated by low CD4(+)-cell count and AIDS diagnosis were predictive of anaemia. CONCLUSION: The results indicate that drug users in the Amsterdam cohort study have a high prevalence of anaemia, which can be explained mainly by high frequency of injecting drugs and HIV infection progression. Reduction or cessation of injecting drug use should reduce anaemia. PMID- 10795360 TI - Diversion of prescribed drugs by drug users in treatment: analysis of the UK market and new data from London. AB - AIMS: To review the available knowledge about the diversion to the illicit market of drugs prescribed to drug users in treatment in the United Kingdom, and to identify aspects of the London market in more detail. METHOD: An analysis of the literature and new data in terms of the extent and nature of the market, the practicalities of trade, motives for selling, reasons for demand and the influence of variations in prescribing practice on diversion. Prices of diverted prescription drugs and details of their availability in London are presented. FINDINGS: The size of the market is substantial and appears to involve a large number of individuals, each diverting small amounts of their own prescribed drugs. Major motives for selling prescribed drugs are to raise funds to buy other, preferred, drugs and/or to pay for a private prescription. Buyers in treatment appear to be motivated by a desire to supplement their own prescriptions because they are dissatisfied with the particular drug prescribed, dosage and formulation. Drug users in treatment can exploit the variations in prescribing practice--such as how much 'take-home' medication they are allowed and whether tests are conducted to ascertain if they are using it themselves--and divert their prescribed drugs. Prices of prescription drugs on the illicit market can fluctuate on a daily basis according to supply and demand. CONCLUSIONS: The results suggest that, to be effective, diversion control must simultaneously involve deterrents from prescribers, drug treatment services, law enforcement agencies and dispensing pharmacists. Finally, some suggestions for further research on this under-studied issue are suggested. PMID- 10795361 TI - The use of antidepressants among injecting drug users in Sydney, Australia. AB - AIMS: To ascertain the prevalence and patterns of antidepressant use among IDU in Sydney and to determine any harm associated with antidepressant use. DESIGN: Cross-sectional survey. SETTING: Sydney, Australia. PARTICIPANTS: Two hundred and one Sydney injecting drug users (IDU) recruited through advertisements, needle exchanges, methadone maintenance clinics and by word of mouth. FINDINGS: Forty per cent of subjects had used antidepressants, 21% in the preceding 6 months. Similar proportions of subjects had used tricyclics (26%) and selective serotonin reuptake inhibitors (SSRIs) (24%), with 8% reporting use of a monoamine oxidase inhibitor. Recent use favoured the SSRIs; however, there was still widespread use of tricyclics. The injection of antidepressants was rare, with only three subjects reporting ever having injected the drugs. Antidepressant use was associated with higher levels of polydrug use, poorer health, higher levels of psychiatric distress and a greater risk of heroin overdose. The excess risk of overdose was specifically associated with tricyclics, rather than SSRIs. CONCLUSIONS: The study confirmed that, like other pharmaceutical products, the use of antidepressants was common among IDU in Sydney. The prescription of tricyclics to heroin users would appear to increase their risk of overdose. If it is considered appropriate to prescribe antidepressants to IDU, it would appear safer to prescribe SSRIs. PMID- 10795362 TI - Consistency of self-administered and interview-based Addiction Severity Index composite scores. AB - AIMS: This study assesses the viability of a self-administered version of the Addiction Severity Index for monitoring substance abuse patients' functioning. DESIGN AND MEASUREMENTS: Patients completed the ASI interview and a self administered questionnaire containing ASI composite items an average of 4 days apart. Composite scores from both formats were compared using correlations and mean differences. PARTICIPANTS AND SETTING: Participants were 316 veterans entering substance abuse treatment in a US Department of Veterans Affairs medical center. FINDINGS: Composite scores for alcohol, drug, psychiatric, family, legal and employment problems correlated 0.59-0.87 across formats. Patients endorsed more drug use and psychiatric symptoms by questionnaire than by interview. Medical composite scores correlated only 0.47 across formats. CONCLUSIONS: This study and previous research suggest that a self-administered questionnaire can be a feasible alternative to ASI interviews for monitoring substance abuse patients' treatment outcomes. PMID- 10795363 TI - A genetic analysis of smoking behavior in family members of older adult males. AB - AIMS: To conduct a genetic study of smoking behavior in 493 three-generation families. DESIGN: Complex segregation analysis and maximum likelihood statistics were used to describe the familial clustering of ever-smoking under several transmission models. SETTING: The Western Collaborative Group Study, an ageing and health study currently in its 39th year of follow-up. PARTICIPANTS: Probands were male participants who were of mean age 71.6 years at the time of the family history interview in 1986-88. MEASUREMENTS: Data were collected via an interview that focused on the family smoking history of participants. Smoking histories of all first-degree relatives were obtained from probands. FINDINGS: Evidence for genetic transmission was indicated by rejection of both the environmental and sporadic models in favor of a Mendelian genetic model with residual familial effects from spouses and both parents. CONCLUSIONS: The best-fitting model was that of a dominant major gene with low estimated frequency and residual familial correlations. This is the first study to date to model the familial transmission of ever-smoking in three-generation families. PMID- 10795365 TI - Recruiting problems for clinical trials. PMID- 10795364 TI - The effects of cigarette consumption on the Sternberg visual memory search paradigm. AB - AIM: To examine the performance of non-smokers (n = 24), light smokers (n = 22, mean 6.5 cigarettes per day) and heavy smokers (n = 19, mean 23 cigarettes per day) on the Sternberg memory search task. DESIGN: A repeated-measures, counterbalanced design was used with one between-subject factor, status (heavy, light or non-smoker) and two within-subject factors, condition (12 hours abstinence or ad libitum smoking) x time (pre- or post-cigarette). FINDINGS: Heavy smokers in the pre-cigarette abstinent session had significantly slower reaction times, movement times and higher intercepts (a measure of factors contributing to performance other than rate of memory scan) than non-smokers. After smoking these differences were removed. CONCLUSIONS: This suggests that rather than improving performance smoking ameliorates a deficit in certain measures of the Sternberg task produced by abstinence. Under ad libitum conditions improvements in performance were attributed to practice. Across all within-subject conditions, there were no significant main effects of smoking status, and this result was consistent with the lack of relationship between measures of saliva continine and expired air carbon monoxide and performance. These data do not support the view that non-abstinent smokers differ from non smokers in the performance of the Sternberg memory search procedure. PMID- 10795366 TI - Meningitis antigen detection: interpretation of agglutination by ultrasound enhanced latex immunoassay. AB - Detailed instructions for performance and interpretation of ultrasound-enhanced latex agglutination tests for the rapid identification of bacteria causing meningitis are described. This recently developed technique, which enhances the sensitivity of most latex immunoagglutination assays, has been studied mainly in the context of detection of antigens of meningitis-causing bacteria. The test concentrates on the Wellcogen bacterial antigen kit (Murex Diagnostics Ltd) that contains five latex suspensions specific for Haemophilus influenzae type b, Neisseria meningitidis ACYW135, N. meningitidis B/Escherichia coli K1, Streptococcus group B and Streptococcus pneumoniae. Light photomicrographs of positive agglutination are shown. Particular attention is paid to the appearance of the latex in negative control samples following exposure to ultrasound. Guidance is given on interpretation and assessment in clinical samples. PMID- 10795367 TI - Release of bacteria during the purge cycles of steam-jacketed sterilizers. AB - The design of the steam-jacketed sterilizer includes an exterior air-gap fixture through which purged chamber aerosols potentially could escape into the ambient environment. Studies of the purge cycle in two sterilizer models tested the potential release of a genetically marked Enterococcus faecalis, together with Bacillus stearothermophilus spores introduced as exposed cultures. Direct plate counts, broth enrichment and polymerase chain reaction analysis were used to confirm any released organisms trapped in an all-glass impinger. From the retrieval of both bacterial strains, an estimated 10(3) organisms can be released from uncontained bacterial loads of 10(11) E. faecalis and 10(7) B. stearothermophilus, even from properly functioning autoclaves. The release of an opportunistic pathogen from sterilizer purge exhausts emphasises the importance of proper sterilizer location, ventilation, containment of heavily contaminated loads, and adequate sterilizer maintenance. PMID- 10795368 TI - Screening for urinary tract infections in a gynaecological setting: validity and cost-effectiveness of reagent strips. AB - This study aims to test the validity and cost-effectiveness of reagent-strip analysis compared with microbiological laboratory testing for mass screening of urine for urinary tract infections in a gynaecological setting. Over a six-month period, urine samples from a convenient group of 228 women presenting in a gynaecological ward of an NHS Trust hospital were tested using Ames 8SG reagent strips to detect leucocyte esterase and nitrite. Total bacterial counts were also carried out; urine culture was recorded as positive if there was > 10(5) organisms/mL. Validity of the dipstick tests was measured, using four criteria (sensitivity, specificity, positive predictive value [PPV] and negative predictive value [NPV]), against the results of laboratory analysis. The combined use of the leucocyte esterase and nitrite tests produced results as follows: sensitivity, 96.4%; specificity, 88.5%; PPV, 54%; and NPV, 99.4%. Cost effectiveness for visible costs was calculated as 48.6% for the 12.3% rate of infection in this study. The use of reagent strips in a mass-screening programme in a gynaecological setting proved both valid and cost-effective. PMID- 10795370 TI - Immunoblot analysis of the antibody response to antigens of Leishmania donovani in Indian kala-azar. AB - When infected with Leishmania donovani, patients develop specific antibodies that constitute the basis of serodiagnosis. Using immunoblot analysis, we examined the antibody response to antigens of L. donovani in 35 kala-azar (KA) patients and 67 controls. Sera from KA patients recognised numerous antigens with molecular weights ranging from 14-110 kDa. Antigens of 40 kDa, 55 kDa, 65 kDa, 70 kDa and 82 kDa were recognised most frequently. All KA patients produce an antibody response to one or more of these antigens. The majority (83%) of KA cases recognised at least four of these five parasite antigens. The 70 kDa antigen showed the greatest sensitivity for Indian KA, and produced a positive reaction in 94% of patients. This antigen gave 10% false-positive reactions in controls comprising patients with related diseases (i.e. tuberculosis, leprosy and malaria) and in healthy controls. Data indicated that the 70 kDa antigen may include a member of the heat shock protein 70 family. Studies with four clinical isolates of L. donovani showed that the 70 kDa component was expressed in all the strains examined. Immunoblot assay (Western blotting) provided a sensitive diagnostic test for KA patients, and identified the 70 kDa parasite antigen that is promising as a potential target antigen for the development of less complex serodiagnostic assays for KA. PMID- 10795369 TI - Familial defective apolipoprotein B-100 (R3500Q) in Northern Ireland. AB - Familial defective apolipoprotein B-100 (FDB) R3500Q is an autosomal co-dominant disorder caused by the substitution of glutamine for arginine at amino acid residue 3500 of the apolipoprotein B-100 gene. It is associated with hypercholesterolaemia of varying severity, and with coronary artery disease. Hypercholesterolaemic patients (n = 158) from Northern Ireland were screened for the defect by polymerase chain reaction-mediated, site-directed mutagenesis. Clinical presentation ranged from moderate hypercholesterolaemia with a family history of hypercholesterolaemia or heart disease (n = 104) to those classified as definitely having familial hypercholesterolaemia (FH) (n = 54). Eight (5.1%) unrelated individuals were found to be heterozygous for the FDB R3500Q mutation, including two (3.7%) of those 54 classified clinically as having FH. Treatment with HMG-CoA-reductase-inhibiting drugs (statins) decreased total cholesterol by 22-44% and low-density lipoprotein cholesterol by 34-46% in all but one FDB heterozygote. PMID- 10795371 TI - Protein C pathway abnormality detection--a national evaluation of three commercial kits. PMID- 10795372 TI - Biological and clinical effects of ozone. Has ozone therapy a future in medicine? AB - Although ozone therapy has been used as an alternative medical approach for four decades, it has encountered scepticism, if not outright objection, by orthodox medicine. This prejudice is not unjustified because ozone therapy often has been used without rational basis or appropriate controls. With the advent of precise medical ozone generators, it is now possible to evaluate some mechanisms of action and possible toxicity. In contrast with the respiratory tract, human blood exposed to appropriate ozone concentrations is able to tame its strong oxidant properties and neither acute nor chronic side effects have ensued in millions of patients treated with ozonated autohaemotherapy. This paper summarises studies aimed at clarifying biological effects, defining any possible damage, the therapeutic window, and suitable doses able to express therapeutic activity. Although an unfashionable and unpopular approach, it is hoped that orthodox medicine will help to critically assess the validity of ozone therapy. PMID- 10795373 TI - Ebola virus. AB - Ebola virus was first identified as a filovirus in 1976, following epidemics of severe haemorrhagic fever in sub-Saharan Africa. Further outbreaks have occurred since, but, despite extensive and continued investigations, the natural reservoir for the virus remains unknown. The mortality rate is high and there is no cure for Ebola virus infection. Molecular technology is proving useful in extending our knowledge of the virus. Identification of the host reservoir, control and prevention of further outbreaks, rapid diagnosis of infection, and vaccine development remain areas of continued interest in the fight against this biosafety level-four pathogen. PMID- 10795374 TI - Facilitative glucose transporter expression in human cancer tissue. AB - Glucose utilisation by cancer cells is greatly enhanced when compared with that by normal tissue. Glucose is taken up by cells and then phosphorylated to glucose 6-phosphate. Facilitative hexose uptake is achieved by five transmembrane transporters, termed glut1-5, which are protein products of their respective GLUT genes. Glut types differ in their kinetics, which are tailored to the requirements of the cell type they serve, although more than one glut may be expressed by a particular cell type. Herein are reviewed the results from approximately 30 studies which examined glut expression in human cancer tissue. These studies measured GLUT messenger RNA (mRNA) either using the reverse transcriptase polymerase chain reaction or by Northern blot analysis, or detected glut proteins using the appropriate antibodies. Tumour tissue is frequently associated with the abnormal and/or over-expression of gluts, especially glut1. Some tumour cells express specific GLUT mRNA but not the respective protein. Some studies have reported associations between glut expression and proliferative indices, whilst others suggest that glut may be of prognostic significance, especially in lung cancer. PMID- 10795375 TI - Dientamoeba fragilis: the unflagellated human flagellate. AB - Dientamoeba fragilis is a pathogenic protozoan parasite with a world-wide distribution. Although originally described as an amoeboid organism, it has been reclassified as a flagellate, on the basis of a number of electron microscopic and immunological findings. Except for its lack of a flagellum, D. fragilis closely resembles Histomonas and Trichomonas. Interestingly, a resistant cyst stage has not been demonstrated and it is unlikely that its trophozoites can survive successfully outside the human host. As a consequence of its higher than anticipated coincidence of infection with Enterobius vermicularis, transmission may occur via ova of this pinworm. D. fragilis infection may be acute or chronic, and has been reported in both children and adults. The most common clinical symptoms include abdominal pain, persistent diarrhoea, loss of appetite, weight loss and flatulence. Occasionally, eosinophilia, urticaria and pruritus have been described. Demonstration of the characteristic nuclear structure of D. fragilis, needed for a definitive diagnosis, cannot be achieved in unstained faecal material; therefore, permanently stained smears are essential. Treatment is recommended in symptomatic cases, and iodoquinol, tetracycline and metronidazole have been used successfully. PMID- 10795376 TI - Choice of controls in the determination of some predictive value parameters. PMID- 10795377 TI - It is often unwise to make predictions about the future. PMID- 10795378 TI - The history of Werner Spalteholz's Handatlas der Anatomie des Menschen. AB - Werner Spalteholz's Handatlas der Anatomie des Menschen is one of the most elegantly illustrated anatomical atlases of all time. Originally published in Leipzig as three volumes from 1895 to 1903, the atlas is still widely used and remains highly regarded by many. The atlas was remarkably popular during the first half of the 20th century, especially the English version in North America and the UK. Unfortunately, the original illustrations and printing plates for the work disappeared following the Second World War and their fate remains a mystery. And, in spite of the atlas's popularity, little is known to the men who prepared the artwork for Spalteholz. It is commonly believed that Max Brodel contributed illustrations to the atlas, but a close examination of the work does not confirm this. A century after its inception, Spalteholz's atlas remains a classic milestone in the history of anatomical illustration. PMID- 10795379 TI - An evaluation of photographic methods to demonstrate the uniformity of sunscreen applied to the skin. AB - Sunscreen is used to help provide some protection from the harmful effects of the sun. Each sunscreen identifies its sun protection factor (SPF) which indicates the degree of protection offered by the product. However, the SPF is dependent upon an internationally agreed amount of 2 mg/cm2 being applied. Studies have shown that people rarely use sufficient sunscreen to offer protection from the sun. A simple photographic technique is described which demonstrates the uniformity of sunscreen applied by users. By employing an ultraviolet fluorescence photographic technique with a sunscreen known to exhibit fluorescent characteristics, it can be shown that in many instances sunscreen is not applied in an even layer. The SPF for the product may therefore not be achieved over the entire area of skin which was intended to be protected by the user. This photographic method could be used in a clinical setting to demonstrate application techniques of skin preparations where uniformity of product on the skin is of utmost importance. PMID- 10795380 TI - Teaching biomedical photographic communications into the next century. AB - The Biomedical Photographic Communications Department of the Rochester Institute of Technology, offers the only Bachelor's Degree in this field in the USA. With over 80 students, the curriculum and its delivery have been continually challenged by the 'New World' of imaging and the demands from the industry where the graduates will work. This paper will examine the programme's recent strategic decisions about curriculum and the future directions for educating tomorrow's professionals. The paper specifically assesses aspects of what is taught, how it is taught, and other relevant issues specific to equipment, networks, ethics, expansion, and extracurricular activities that have been put in place. PMID- 10795381 TI - Digital data and the 19th century teratology collection. AB - The golden age of descriptive teratology (congenital anomalies) was between 1750 and 1850. During that period, the study of human congenital malformations, especially those dramatic examples designated as 'monsters', attracted special attention. One of the finest collections in this field was Museum Vrolikianum, the collection of father and son, Gerardus (1775-1859) and Willem Vrolik (1801 1863). Willem, a specialist in teratology, described many of these specimens in his Handbook of Pathological Anatomy (1844) and his teratology atlas Tabulae ad illustrandam embryogenesin hominis et mammaliam tam naturalem quam abnormem (1849). This collection is still an integral part of the Museum Vrolik in the Academic Medical Center in Amsterdam. From 1991 to 1994, the collection of congenital anomalies was re-catalogued and re-described according to contemporary syndromological views. We diagnosed rare syndromes with multiple congenital anomalies, rare skeletal dysplasias, closure defects of the neural tube and conjoined and acardiac twins. We came to the conclusion that some anomalies of the conditions diagnosed have been reported only a few times. Certain cases demonstrate relationships between different malformations, and provide critical 'missing links' in the teratological series. For the diagnosis of the clinical syndromes in some of the specimens magnetic resonance imaging (MRI), computed tomography (CT), spiral CT and radiographical techniques were of crucial importance and are discussed in this paper. PMID- 10795383 TI - Occupational medicine in the new millennium. PMID- 10795382 TI - How the idea of profession changed the writing of medical history. PMID- 10795384 TI - Physical activity, fitness and body composition of Finnish police officers: a 15 year follow-up study. AB - This study evaluated changes in the physical activity, fitness and body composition of 103 police officers during a 15-year follow-up. The absolute aerobic capacity was similar in 1981 and 1996, muscular performance had declined, and body weight had increased approximately 0.5 kg/year. More than half the subjects (53%) had increased their leisure-time physical activity in 1996. The correlation was significant between physical activity in 1981 and physical fitness in 1996, but weak between physical activity in 1996 and fitness in 1996. It was also significant between waist circumference and waist/hip ratio in 1996 and physical activity during the previous 5 and 15 years. No significant correlations were found between physical activity and work ability or perceived physical or mental job stress. The physical fitness of middle-aged police officers seems to be predicted strongly by physical activity in early adulthood. Therefore health and fitness promotion measures should start at that time. This, together with regular systematic training, should help to sustain work ability of middle-aged police officers. PMID- 10795386 TI - Variation in licensing authority standards for diabetic taxi drivers in the United Kingdom. Driving and Employment Working Party of the British Diabetic Association. AB - In the UK, prospective taxi-drivers with diabetes are licensed locally according to variable criteria. In order to determine the criteria used for licensing, questionnaires were sent to all 422 local licensing authorities. Responses were obtained from 372 authorities (88% response rate). Only 201 (54%) currently issue licences to drivers on insulin treatment, and 260 (70%) to those on diet and/or oral hypoglycaemic agents. Most local authorities would prefer taxi-licensing to operate centrally rather than locally. It is apparent that the current UK system for licensing diabetic taxi drivers is variable and not evidence-based. A nationally organized system with clearly defined criteria and the capacity for individual consideration, would be more fair and consistent, and would be unlikely to compromise road safety. PMID- 10795385 TI - Increased risk of coronary heart disease in Japanese blue-collar workers. AB - The purpose of this study was to test the hypothesis that blue-collar workers have an increased risk of coronary heart disease. Subjects were 653 male (mean age, 32.6 years) and 492 female (mean age, 26.3 years) workers employed during 1993 at a computer manufacturing company. The measurements determined were body mass index (BMI), waist to hip ratio (WHR), skinfold thickness, blood pressure, grip strength, back strength, vertical jump and trunk flexion. Male blue-collar workers had significantly higher WHR (P < 0.001), systolic blood pressure (P = 0.031), grip strength (P = 0.030), and standing flexion (P = 0.038) than do white collar workers. Female blue-collar workers had significantly higher BMI (P = 0.018), WHR (P < 0.001), and grip strength (P = 0.007). Blue-collar workers took significantly less exercise, and habitually snacked (females only), smoked and drank more alcohol (males only) than white-collar workers. In conclusion, blue collar workers have more intra-abdominal fat than white-collar workers of either sex. The difference may be due to factors contributing to the less healthy lifestyles of blue-collar workers. PMID- 10795387 TI - Occupational histamine poisoning by fish flour: a case report. AB - Histamine poisoning due to inhalation and skin contact with fish products is rarely described in the literature. This study presents a case of occupational histamine poisoning by spoiled fish flour via inhalation, skin and eye contact. Shipments of fish flour transported in black or blue bags, depending on the source, were handled by 20 harbour workers. Ten workers handling blue bags developed allergy-like skin, eye, gastrointestinal, respiratory and cardiac symptoms within 30 min. Workers handling black bags were symptom-free, except for minimal eye irritation. After consultation with the Poison Control Centre histamine poisoning was suspected. The histamine content, as determined by thin layer chromatography, was 10-fold higher in samples from the blue than from the black bags (510 mg/100 g flour compared with 50 mg/100 g flour, respectively). Part of the shipment was labelled as hazardous for human health with permission for further usage only under specific personal protective measures. It is suggested that the highest permissible levels of histamine in fish flour and similar products should be set and legally adopted. PMID- 10795388 TI - Respiratory symptoms and wheat flour exposure: a study of flour millers. AB - The objective of this study was to determine the prevalence of respiratory symptoms and their relationship to sensitization to wheat flour allergens and fungal amylase, in a group of workers from the UK flour milling industry. A cross sectional study was used to evaluate symptoms, using a structured interview technique, and sensitization, using skin prick test findings, from 679 employees in flour milling and packing operations at 18 flour mills. A total of 147 workers (147/679, 22%) described upper respiratory tract symptoms of some kind. In the majority (139/147, 95%) these symptoms were of an occasional or transitory nature and were related to short-term exposures to high levels of dust. Three individuals (3/679, 0.4%) were identified whose symptoms were thought to be the result of allergy to wheat flour. The prevalence of positive skin prick tests to wheat flour allergens and to fungal amylase was 1.2% (8/678) and 0.9% (6/678), respectively. Measurements of total inhalable dust exposure for personnel exposed to flour dust were taken at 10 of the sites (116 samples). For production personnel the levels were typically between 1 and 10 mg/m3 (8-h time weighted average), with a median of 6.2 mg/m3. Hygiene operatives had appreciably higher exposure than production personnel, with a median of 18.7 mg/m3. Overall, 43% (50/116) of all samples exceeded 10 mg/m3 (8-h time weighted average). The findings suggest that the risk of sensitization to wheat flour allergens from current dust exposures in flour mills is very low. Wheat allergy was responsible in only a small proportion of the total who had respiratory symptoms. The principal causation of symptoms experienced by the workforce was considered to be a non-specific irritant effect related to short-term exposures to high levels of total inhalable dust. PMID- 10795389 TI - Sensory neural processing in work-related upper limb disorders. AB - Pain in the upper limb is a common complaint in adults, and is often attributed to or exacerbated by occupational activities. In many patients there is no demonstrable pathology in the neck or arm to account for the symptom, and this has prompted the hypothesis that such cases might arise through abnormal neural processing of sensory information with a lowering of pain thresholds. In this paper we review the evidence in support of this theory and suggest directions for future research. PMID- 10795390 TI - The development of a health policy for the Army. AB - This paper describes the development of the Army Health Policy (AHP), which is a key component of the Army Human Resources Strategy (AHRS). The work on the AHP provided an opportunity for a fundamental review of the delivery of health support to the Army. The AHP will provide the strategic framework by which the Army will ensure the health of its workforce and, where appropriate, their dependents. The methodology used for this work may be a useful model for the development of a health policy for occupational populations. PMID- 10795391 TI - Do symptomatic radiographers provide evidence for 'darkroom disease'? AB - Non-controlled studies performed in New Zealand and the UK reported a high incidence of a large number of symptoms within radiographers. The authors of these investigations proposed that these symptoms resulted from exposure to X-ray processing chemicals and the associated fumes, and collectively labelled the condition 'darkroom disease'. The absence of control groups in these investigations weakens the 'darkroom disease' hypothesis. The current work set out to address this deficiency. Forty percent of radiographers (n = 295) and 40% of physiotherapists (n = 250) working in 34% of Irish hospitals (n = 31) were asked if they experienced any of the 15 symptoms described in 'darkroom disease'. The results demonstrated that radiographers had a significantly higher incidence than physiotherapists for only two of the symptoms--bad taste (P < 0.0001) and sore eyes (P < 0.001). These higher incidences were confined to three of the 31 hospitals surveyed. Physiotherapists expressed a higher incidence for sore throats (P < 0.01) and nasal discharge (P < 0.01). These results clearly demonstrate that radiographers are no more symptomatic than a group of hospital staff not exposed to processing chemicals. The current findings offer no support for the 'darkroom disease' hypothesis. PMID- 10795392 TI - Cardiovascular effects of occupational exposure to carbon disulphide. AB - The objective of this study was to investigate the effect of occupational exposure to carbon disulphide (CS2) on the total cholesterol, blood pressure and prevalence of coronary heart disease (CHD). A cross-sectional study involving 252 viscose rayon workers and 252 age and sex matched controls was carried out. Depending on the job and specific work place, the CS2 concentrations were between 10 and 64 mg/m3. A cumulative exposure index (CS2 index) was calculated for each worker by multiplying the number of years he had held a particular job with the CS2 concentrations in that job. CHD prevalence among the exposed was higher than among the controls; the difference reaching significance only for highly exposed workers. Cholesterol levels were significantly higher in both highly and moderately exposed groups. In conclusion, the results demonstrated that occupational exposure to CS2 increases total cholesterol and the risk for CHD. While the risk for CHD is increased in workers exposed to high CS2 concentration for many years (CS2 index > or = 300), even the relatively modest exposure (CS2 < 300) may increase the serum cholesterol. PMID- 10795393 TI - Effects of exposure to very high frequency radiofrequency radiation on six antenna engineers in two separate incidents. AB - Six men are likely to have been accidentally exposed to high levels of very high frequency (VHF) radiofrequency radiation (100 MHz) while working on transmission masts; four men in one incident and two in another. They experienced symptoms and signs which included headache, parasthesiae, diarrhoea, malaise and lassitude. The condition of four men, two men from each incident likely to have had the highest exposure, has shown no significant improvement. The first incident occurred in 1995 and the second in 1996. PMID- 10795394 TI - Partnerships and joined-up working: the millennium key? AB - Fresh challenges for occupational health in the new millennium include the European dimension and a series of frame shifts about what our specialty actually comprises. This paper encourages a proactive commitment to engaging with Europe and European law-making. A similar proactive approach involving professional, employer and workforce participation is urged for the development of occupational health. The need for prioritization and economic justification is required for underpinning this approach. PMID- 10795395 TI - The filling of drinking vessels at social events: courtesy versus ergonomics? PMID- 10795396 TI - Should control measures be based on air measurements or biological/biological effect monitoring? AB - The setting of, and the review of, exposure limits takes into account toxicological, occupational hygiene and epidemiological data. The COSHH Regulations 1994 define a hierarchical approach to controlling workplace exposures with a particular emphasis on the measurement and control of airborne substances. Absorption via the lungs is considered the most important route of entry in the workplace, however, percutaneous absorption must not be overlooked. Biomarkers are used extensively in the surveillance of workers' exposure to metals and organic chemicals. In addition, Genetic polymorphism for xenobiotic metabolism has been widely studied. The selection, validation and application of any biomarker is a complicated process and requires careful consideration prior to any application. PMID- 10795397 TI - A South-East Asian perspective. AB - In order to discuss the subject of occupational medicine in the next century, changes in the present demographic profile and work activity must be considered first. Only then can the challenges be identified, and appropriate strategies be formulated to respond to them. In the diverse countries of South-East Asia, improved health and work conditions, the advent of new technology, a redistribution of work activity, and an ageing workforce can be expected. Two other factors that have specific impact in the region are the recent financial crisis and the occurrence of an international environmental haze from forest fires. The various countries in South-East Asia, which are in different stages of development, and have different problems and priorities, will respond differently to the demands for occupational health. It is likely that there will be a shift in the focus of current health care activities towards specific work sectors, the recognition of new hazards at work, the identification of newly emerging work related diseases, and an increase in health promotion in the workplace. Hopefully, there will be improved training of health professionals to ensure that there are adequate numbers and that they are well prepared to face these changes. Responsive, appropriate and well enforced labour legislation to protect the health of all workers, and international cooperation in occupational and environmental health are also required. As global and regional economic conditions continue to remain unstable and the impact of the crisis further takes its course, the final effect on occupational health in South-East Asia remains to be seen. PMID- 10795398 TI - Modern view of vitamin D3 and its medicinal uses. PMID- 10795399 TI - Neurokinin receptor antagonists. PMID- 10795400 TI - Opioid receptor antagonists. PMID- 10795401 TI - Mechanisms of bacterial resistance to antibiotics and biocides. PMID- 10795402 TI - Towards cannabinoid drugs--revisited. PMID- 10795403 TI - Is it a modacrylic fibre? AB - This study describes the characteristics of modacrylic fibres and includes over 80 samples (previous and current) representing 15 trade names. Fibre morphology was examined using brightfield microscopy. Signs of elongation were determined using polarised light microscopy. Fibre cross sections were also examined. The generic class of fibre was divided into sub groups using polymer composition as determined by FTIR-microscopy. Microscopically, some modacrylic fibres cannot be distinguished from acrylic fibres. Others display unusual optical and morphological features which are a strong indication of their generic class. The infrared spectra provide information about the co-monomer, termonomers added to produced dye sites, the presence of solvent residue, dyes, and additives, e.g. flame retardant material. The infrared spectra should always be recorded before and after any thin layer chromatographic examination of the dye, otherwise peaks attributable to dyes, which may be a valuable comparative feature in casework will be lost. PMID- 10795404 TI - A 'cyanoacrylate case' for developing fingerprints in cars. AB - A portable case has been developed by which cyanoacrylate (super glue) fuming can be used inside a vehicle suspected of being involved in serious crime. The car itself serves as a fumigation chamber and the cyanoacrylate vapours are fed into the car via a hose. Connected to the hose and suspended inside the car is a vapour diffuser. The cyanoacrylate originates from a portable case where there is a sealed heater and also a command panel with hygrometer and thermometer for a technician to control the process. There is also space inside the case for other necessary equipment. PMID- 10795405 TI - Individual identification of cats and dogs using mitochondrial DNA tandem repeats? AB - Cats and dogs are very common animals in the human environment. In Switzerland, one in five households owns a cat or a dog. Their hairs are very easily transferred and could be used as a frequent trace evidence. Using DNA analysis, identification of these animals should be possible as it is in human identification. However, most of the time, no nuclear DNA can be recovered from the hair. It is therefore necessary to rely on mtDNA. Cats and dogs have tandemly repeated sequences in their mtDNA control region. In this study, the authors show that these tandem repeats are very polymorphic but are also the source of a very high level of heteroplasmy. The authors, therefore, examined if this might prevent their use in forensic identification. PMID- 10795406 TI - Minimum lung function for breath alcohol testing using the Lion Alcolmeter SD 400. AB - The aim of this study was to determine the minimum lung function required by an individual to complete a breath alcohol test successfully using a Lion Alcolmeter SD-400. A total of 331 subjects routinely referred to a busy clinical respiratory function laboratory for assessment of their lung function were tested. A total of 52 (15.7%) of these subjects were unable to provide an adequate sample after two attempts. The majority of the group (86.5%) were female. There was considerable overlap of the minimum lung function parameters between those subjects who were unsuccessful at all attempts and those who were successful. The failure rate of the subjects using the Lion Alcolmeter SD-400 was approximately 50% of that found in a previous study of the Lion SD-2. PMID- 10795407 TI - Unusual results due to transfused blood. AB - During a murder enquiry, the forensic science investigation used PGM and EAP blood grouping systems and detected a mixture of blood on the deceased's jacket. The blood groups matched those of the deceased and accused. The results of DNA analysis, however, proved that only a single source of DNA, matching the deceased, was present. Supplementary information relating to the transfusion of the individual whilst still wearing his clothing led the authors to conclude that a mixture of transfused blood and the individual's own blood had effused from his body via a stab wound, and onto his clothing. PMID- 10795408 TI - Identification of organic pigments in coatings: applications to red automotive topcoats. Part III: Raman spectroscopy (NIR FT-Raman). PMID- 10795409 TI - The examination of UV-absorbers in 2-coat metallic and non-metallic automotive paints. PMID- 10795410 TI - Recapturing the essence of criminalistics. PMID- 10795411 TI - The likelihood approach to compare populations: a study on DNA evidence and pitfalls of intuitions. AB - The paper follows on from earlier work [Taroni F and Aitken CGG. Probabilistic reasoning in the law, Part 1: assessment of probabilities and explanation of the value of DNA evidence. Science & Justice 1998; 38: 165-177]. Different explanations of the value of DNA evidence were presented to students from two schools of forensic science and to members of fifteen laboratories all around the world. The responses were divided into two groups; those which came from a school or laboratory identified as Bayesian and those which came from a school or laboratory identified as non-Bayesian. The paper analyses these responses using a likelihood approach. This approach is more consistent with a Bayesian analysis than one based on a frequentist approach, as was reported by Taroni F and Aitken CGG. [Probabilistic reasoning in the law, Part 1: assessment of probabilities and explanation of the value of DNA evidence] in Science & Justice 1998. PMID- 10795412 TI - Paternity probability when a relative of the father is an alleged father. AB - When scientists use DNA evidence in court, coancestry effects such as population structure and relatedness are usually ignored. In paternity cases, only if a particular man has the child's paternal allele at a certain locus, can he not be excluded in the paternity dispute. However, it is certainly true that close relatives will be far more likely to have the child's paternal allele than will random members of the reference population. In particular, the probability that the true father's brother has the paternal allele is very much greater than that for any other relationship. In this paper, the authors describe a method for inference in a case where the true father may be a relative of the alleged father. This paper also reports that most current methods overstate the probability that the alleged father is the father. PMID- 10795414 TI - The transfer and persistence of automotive carpet fibres on shoe soles. AB - The transfer and persistence of automotive carpet fibres to shoe soles was investigated. It was found that fibres were transferred with the normal activity of a car passenger. Carpet type and shoe sole parameters were significant determinants in the number of fibres that transferred. The average number of fibres was between about one and 33 per sole. Fibres that had been transferred after normal activity only persisted for a few minutes after walking. A survey of the shoe soles of people about to leave their car showed that fibres were usually present. The majority of shoe soles surveyed had less than five fibres with the greatest number of fibres found being 14. The likelihood of finding a large number of fibres on such soles is rare. Fibre composition of automotive carpets showed a high degree of variation. Grey was seen to be a common colour irrespective of the colour of the vehicle body. PMID- 10795413 TI - The identification of credit card encoders by hierarchical cluster analysis of the jitters of magnetic stripes. AB - The relative bit density variation graphs of 207 specimen credit cards processed by 12 encoding machines were examined first visually, and then classified by means of hierarchical cluster analysis. Twenty-nine credit cards being treated as 'questioned' samples were tested by way of cluster analysis against 'controls' derived from known encoders. It was found that hierarchical cluster analysis provided a high accuracy of identification with all 29 'questioned' samples classified correctly. On the other hand, although visual comparison of jitter graphs was less discriminating, it was nevertheless capable of giving a reasonably accurate result. PMID- 10795415 TI - A method for the rapid detection of the zopiclone degradation product 2-amino-5 chloropyridine. AB - Zopiclone (Zimovane) is a cyclopyrrolone compound which exhibits hypnotic and sedative effects while also exhibiting anticonvulsant and muscle relaxant activities. The detection and quantification of zopiclone is difficult. It has a high molecular weight compared to most other commonly used drugs, therapeutic levels are not high, and it is unstable in nucleophilic solvents. A degradation product of zopiclone, 2-amino-5-chloropyridine (ACP) together with a method for its detection using high-performance liquid chromatography with diode array detection has been described previously. An account is presented of a simple method for the detection of ACP using gas chromatography with mass selective detection (GC/MS) which will facilitate detection of zopiclone use as part of a routine screen. PMID- 10795417 TI - Workshop on explosives trace analysis methods. PMID- 10795416 TI - When can a DNA profile be regarded as unique? AB - The probability that a defendant's DNA profile is unique in a population of untyped individuals is shown to be bounded below by one minus twice the sum of the match probabilities over the population. This bound assumes that the possibility of laboratory or handling error can be neglected, and applies only when there is no non-DNA evidence in favour of the defendant. There cannot be a completely general lower bound: if there is overwhelming non-DNA evidence that the defendant is not the source of the crime stain, then that is also overwhelming evidence of non-uniqueness. Application to k-locus short tandem repeat (STR) profiles is discussed, and illustrated with calculations based on the 6-STR-locus system used in current UK casework. However, because of the problem of the non-DNA evidence, there seems to be no satisfactory way for an expert witness to address the question of uniqueness in court. PMID- 10795418 TI - Fibre comparisons using microspectrophotometry. PMID- 10795419 TI - Amount of blood needed for DNA testing. PMID- 10795420 TI - Forensic science accreditation and training. PMID- 10795421 TI - Wizards and gatekeepers at the roadside? PMID- 10795422 TI - More on the hierarchy of propositions: exploring the distinction between explanations and propositions. AB - Interpretation of the weight of scientific evidence depends upon the framing of at least two competing propositions to weigh against each other. It is the stage of framing propositions that is the most difficult aspect of evidence interpretation. The logical structure for case assessment and interpretation has been described by the authors in three previous papers [Cook R, et al. A model for case assessment and interpretation. Science & Justice 1998; 38: 151-156. Cook R, et al. A hierarchy of propositions: deciding which level to address in casework. Science & Justice 1998; 38: 231-239. Cook R, et al. Case pre-assessment and review in a two-way transfer case. Science & Justice 1999; 39: 103-111]. This paper considers the framing of propositions in greater detail, in particular the intermediate stage of exploring less formal explanations. All of the discussion is based on experiences encountered in workshops with caseworking forensic scientists. PMID- 10795423 TI - The detection and persistence of Cannabis sativa DNA on skin. AB - The presence of Cannabis sativa DNA was detected on the skin of persons who have recently handled both leaf and resinous material. The persistence of C. sativa DNA was examined on the skin. The subjects were asked to either repeatedly rub their hands on their trousers, place their hands repeatedly into their pockets or wash their hands in soap and water. After rubbing the hands on trousers or placing them in pockets C. sativa DNA could still be detected. No DNA could be detected after washing the hands. PMID- 10795424 TI - The biodegradation of the fabric of soldiers' uniforms. AB - A study to determine the changes in the morphological structure of woollen fabric and fibres of military uniforms, after burial in biologically active soil for a particular period of time. The analysis of fabric and fibres was performed by means of organoleptic and microscopic observations. The final decay of the fabric of soldiers' uniforms took place after 5-6 weeks burial in the standardised bioactive soil. The changes of the morphological structure of fabric undergoing the process of biodegradation depended mainly on the duration of their burial in the soil. PMID- 10795425 TI - Death and injury caused by land mines in Burma. AB - One hundred and eighty-eight Burmese refugees in Thailand were interviewed. One hundred and five of those interviewed had knowledge of a total of 313 persons who had been exposed to land mine explosions. Twenty-three of the interviewed were land mine survivors. They were all male, aged between eight and 68 years, and all except one had been injured between 1986 and 1997. Fifteen of the 23 were civilians, eight were guerilla soldiers who were injured either in battle or while deploying or disarming land mines. Those who had stepped on land mines were all crus or femur amputated and had several scars on the lower limbs, abdomen and some also on the forearms. Those who had handled a land mine had lost either fingers or hands and one also lost both eyes. This study shows that the frequency of land mine accidents in Burma has been underestimated and that the mortality is high. The study also supports the general impression that many victims are civilians. PMID- 10795426 TI - Evaluating the statistical significance of single band profiles in VNTR analyses. AB - VNTR profiles may present either a single band or two bands. If two bands are present then the individual is a heterozygote for these two bands. However, if only one band is present there is ambiguity as to the true genotype of the individual. This person may be a homozygote in that he has two copies of the same allele, or he may be a heterozygote for two very close bands that cannot be separated on the gel. The second NRC report proposed the use of the '2p' rule, or Formula 4.10a in the sub-structure case, as a conservative upper bound in the statistical interpretation. However, further examination suggests that these formulae are not necessarily conservative. In this paper we examine this phenomenon by deriving a formula that contains both the corrections for null alleles and for subpopulation effects. PMID- 10795427 TI - The calculation of DNA match probabilities in mixed race populations. AB - A coherent method is offered to estimate likelihood ratios for DNA match probabilities from mixed racial populations that avoids the approach of reporting separate estimates for each race. The method is demonstrated for some cases involving profiles derived from several individuals and incorporates a correction for 'subpopulation' effects. PMID- 10795428 TI - Can wine tasting be used as a defence to a charge of excess alcohol? AB - Wine-tasting has been used as a defence in the UK to charges of driving with excess alcohol on the breath. There is little scientific data to support this explanation. The authors carried out an experimental wine-tasting using twelve volunteer subjects. Breath alcohol samples were taken at the start of the experiment and fifteen minutes after the end. All subjects after wine-tasting had breath alcohol levels below the instrumental limit of detection. The authors conclude that the wine-tasting defence is not credible. PMID- 10795429 TI - The diagnostic significance of haemorrhage of the posterior cricoarytenoid muscles. AB - A number of authors have indicated that haemorrhage of the posterior cricoarytenoid (PCA) muscles can be used as a diagnostic criterion to establish the cause of death. Other authors even consider it to be 'forensic evidence' when differentiating between natural and violent death. They agree on agonal asphyxia or dyspnoea as a developmental mechanism, possibly in combination with a 'convulsive' dilatation of the glottic cleft. In view of this difference of opinion, we conducted a study of our own. Retrospective evaluation of 2060 post mortem examinations performed in 1996 at the Institute of Forensic Medicine of the University of Munich disclosed 28 cases demonstrating visible haemorrhage of the PCA muscles. The study revealed no correlation between these findings and certain causes of death, nor any indication of certain mechanisms of development. PMID- 10795430 TI - Note on fabric marks in motor vehicle collisions. AB - Two cases are described where motor vehicle accidents lead to fabric weave impressions and fibres being transferred to the paintwork of vehicles from the clothes of the victims. PMID- 10795431 TI - [How could we improve the results of our health counseling?]. PMID- 10795432 TI - [Coronary risk and prescription in primary care patients with hypercholesterolemia]. AB - OBJECTIVE: In patients with hypercholesterolaemia determinate the prevalence of high coronary risk (CR), study the lipid lowering treatment applied and determinate if there is any change in CR after a period of treatment. DESIGN: Cross-sectional. EMPLACEMENT: Primary care. PATIENTS: 583 patients with hypercholesterolaemia both sex, older than 25 years registered in chronic mobility, randomized selected. MEASUREMENT AND RESULTS: Applying the Framingham coronary multivariate risk method we estimate high CR > 20%. Patients with a previous history of cardiovascular event, were treated in a 50%, more frequently younger subjects, rising 220 mg/dl of final cholesterol level. Patients without any cardiovascular event known, the 32.5% (28.0-36.7%) have a CR > 20%. Subjects with high CR have 4.9 (3.0-8.2) more probability if receiving treatment than the others with lower risk. The lipid-lowering treatment is explained in a 67% because the high CR and the family history of coronary event. After at least one year period there is a reduction in those with high CR (difference relative of proportions 28.7% [20.4-37.1]). PMID- 10795433 TI - [The debate on the organization, functions and efficiency of nursing in primary care: apropos of a qualitative study]. AB - OBJECTIVE: To show the opinions that doctors and nurses express on the organisation and competences of primary care nursing, and on inter-professional relations. DESIGN: Qualitative research study based on the group discussion technique known as Philips 66. SETTING AND PARTICIPANTS: Three discussion groups, one of nurses, one of doctors and a third mixed, with 6 components each, half from Caceres and the rest from other provinces, and all involved in primary care health delivery. MEASUREMENTS AND MAIN RESULTS: In the discussions, the three groups coincided in the importance given to nursing clinics and home visits, in the organisational advantages that the attachment as a norm of the population to a nursing clinic would bring, in the ability of nurses to perform the techniques they have habitually used, and in defence of one sole clinical record per patient. They blamed each other mutually for making few home visits. Doctors attributed to nurses under-use of records and nurses alleged doctors took no notice of their notes. On the question of their relations, doctors thought that nurses did not take on joint responsibility for the work-loads in health centres; and nurses thought that doctors did not wish to share tasks, but rather delegate or order them. CONCLUSIONS: The attachment of the population to nursing clinics would favour the extension of care, the commitment to the population and the real delivery of services, as well as professional autonomy and recognition. PMID- 10795434 TI - [Evolution of mortality due to lung cancer in Andalusia between 1975 and 1977]. AB - OBJECTIVE: To describe mortality due to lung cancer in those over 24 by sex, age, birth cohort and year of death in Andalusia between 1975 and 1997. DESIGN: A descriptive population-based study. SETTING: Andalusia between 1975 and 1997. PARTICIPANTS: All those residing in Andalusia during the study period. MEASUREMENTS AND MAIN RESULTS: The following indicators were calculated: gross rates, rates adjusted by age of death and rates of potential years of life lost. The European population was used as reference, with specific rates for age-groups and year of birth. Poisson models were used to quantify trends and annual percentages of change both for the entire period and for the 23 years studied. Death from lung cancer in men during these years increased markedly. Age-adjusted rates went up from 77.37 deaths per 100,000 persons per year in 1975 to 121.92 in 1997. However, in women, there was a small drop from 10.19 to 8.22 deaths per 100,000 persons per year over the same period. Men suffered 7.59 times more mortality than women in 1975, reaching 17.25 times more in 1994. Age had a clear relationship to lung cancer mortality: the older the population, the greater the death-rate. In men there was an ascendant cohort effect until the generation born between 1950 and 1959, from which time the effect is unclear. Women had a descending cohort effect, with high variability between one generation and another. CONCLUSIONS: Death from lung cancer in Andalusia between 1975 and 1997 increased in men in transversal terms, whereas in women it went down. However, the analyses of birth cohorts show signs of changes in the most recent generations, although age, in both men and women, is the variable most closely related to mortality. PMID- 10795435 TI - [Depression in the elderly and the role of family dynamics]. AB - OBJECTIVE: The objective was to find the relationship between the lived experience within the family and depression in the elderly. DESIGN: The study had a crossover and descriptive design. SETTING: Carranque and Campanillas Health Districts, Malaga. PATIENTS: The target population was the over 65's. The sample consisted of 97 people attended at 2 health centres, one rural and one urban. MEASUREMENTS AND MAIN RESULTS: The main study variable was the results on the Yesavage Geriatric Depression Scale (GDS), which were contrasted through multiple linear regression with the variables of age, sex, marital status, tobacco dependency, number of people living together, presence of the children in the home (concretely, daughters), family income, ownership of the home, social support network, and Apgar Family II. The results showed that the GDS correlated only with the results of the Apgar Family II (p = 0.00003). CONCLUSIONS: When tackling the depression of elderly people in primary care, it is absolutely vital to look at the family context, as this is a factor strongly linked to the depression. PMID- 10795436 TI - [Anemia in primary care: etiology and morphological characteristics]. AB - OBJECTIVE: To find the morphological characteristics and causes of the types of anaemia seen at a primary care centre. DESIGN: Descriptive, observational study. SETTING: Urban health centre. PATIENTS: People attending for a year who had an anaemia defined by haemoglobin figures below 13 g/dl in males and 12 g/dl in women. MEASUREMENTS AND MAIN RESULTS: 152 patients with anaemia were identified. The most common types of anaemia were iron-deficiency anaemia (IDA), anaemia due to chronic illness (ACI) and post-haemorrhage anaemia (48%, 26.3% and 6.6%, respectively). Anaemia due to vitamin B12 deficit was detected in four patients, Thalassaemia minor in two, haemolytic anaemia in two, and a refractory anaemia in one patient. The most common cause of IDA was gynaecological in origin; and the commonest cause of ACI was neoplasm. The main findings of digestive origin in IDA were oesophagitis in two patients, duodenal ulcer in one, erosive gastritis in one, gastric neoplasm in one, colonic neoplasm in two and Crohn's disease in one. 13.7% of the anaemia studied in PC required hospital referral. CONCLUSIONS: Anaemia is a common health problem in primary care (PC), with a rough incidence of one case per month per doctor. Its main types are iron-deficiency anaemia and anaemia due to chronic illness. Most cases were detected in PC and most can be studied properly at this care level. PMID- 10795437 TI - [Impact of various objectives related to the rational use of medication in a primary care area]. AB - OBJECTIVES: To evaluate the impact of the application of objectives relating to the rational use of medication and to assess the possible effect of these objectives on pharmaceutical expenditure. DESIGN: A non-randomised intervention study with control. SETTING: Santiago de Compostela Primary Care (PC) Area. PATIENTS AND OTHER PARTICIPANTS: After the inclusion and exclusion criteria were applied, both the intervention group (IG) and the control group (CG) consisted of 31 doctors. Those in the IG belonged to the reformed PC model, whereas those in the CG did not. INTERVENTIONS: Four blocks of objectives on the rational use of medication were included in the Pact signed by reformed-model doctors and the management. The study lasted three years (1996-1998) for the objective of lowering the prescription of low-therapeutic-utility (LTU) drugs, and two years (1997-1998) for lowering the prescription of antihypertensives, Triflusal and quinolones. MEASUREMENTS AND MAIN RESULTS: The difference between both groups was 1.7 fewer packs of LTU drugs per 1000 users and per day in the IG (95% CI, 0.9 2.6) and 1768 pesetas less (95% CI, 801-2733). These differences were still greater among pensioners. The difference between the two groups in terms of total pharmaceutical cost was 7379 pesetas less in the IG (95% CI, 708-14,049) than in the CG. The differences between the groups for the remaining objectives (antihypertensives, Triflusal and quinolones) in no case reached statistical significance. CONCLUSIONS: The pharmaceutical objectives of the pact were partly achieved: a drop in LTU drug prescription was seen, with an indirect reduction in overall expenditure, but no differences were found in the prescription of antihypertensives, triflusal or quinolones. PMID- 10795438 TI - [Health education in schools for adults: by a teacher or health education lecture?]. AB - OBJECTIVE: To compare the results of two ways of teaching the rational use of medicines to students of centres of permanent education of adults (CPEA): one taught by the normal teachers (after training by health personnel) and one through a lecture given by the health staff. DESIGN: Intervention study without randomised distribution and with a control group. SETTING: Five CPEA in an urban centre. PARTICIPANTS: 385 students and 15 CPEA teachers. INTERVENTIONS: Three groups: a) "teachers" group: consisting of students who received education on medicines in the class-room through their teachers, who had been previously trained by health personnel; b) "lecture" group: students who had received a health education lecture on medicines given by health staff; c) non-intervention group. All three groups were administered a questionnaire before and after the intervention. Both questionnaires were paired. MEASUREMENTS AND MAIN RESULTS: 248 people completed the first questionnaire and 149 the second. Significant gains in knowledge were only found in the teachers intervention group (p < 0.01; 7.8% increase in score). Dividing the students into terciles made these gains significantly greater (11.7%) in the students of the teachers group who in the first questionnaire had intermediate scores than in the students in the other groups who had intermediate scores. CONCLUSIONS: Intervention with teachers seems more effective than either a health education lecture or no intervention, especially in the improvement in knowledge of students who already had beforehand intermediate knowledge. PMID- 10795439 TI - [Domiciliary nursing care in a special emergency service: characteristics of the population attended]. AB - OBJECTIVE: To analyse the characteristics of the users of programmed home nursing care in the Special Emergency Service (SES) in Zaragoza. DESIGN: Cross-sectional, descriptive study. PATIENTS: Patients who asked for programmed home nursing care on 5 consecutive sundays of each quarter of 1998. SETTING: SES in Zaragoza. MEASUREMENTS AND MAIN RESULTS: We analysed the characteristics of the SES users as to age and sex, origin by health districts and areas, and nursing services requested. 1104 of the 3507 calls made were from men (31.5%), and 2403 women (68.5%). Average age was 73.21 (+/- 13.54). Services most in demand were the administration of anti-clotting agents (31.46%), followed by healing treatment (19.44%), mainly for bedsores. Area II had a greater rate of attendance (6.28%), with more demand from the four consulting clinics belonging to this area than from the health centres. CONCLUSIONS: The general profile of the patient at home is of women with an average age of 74.27 (+/- 12.77), who are administered anti clotting drugs or receive care for sores. These require primary care nursing attention to improve their quality of life and satisfaction. There were differences in the rates of demand between the different health areas. PMID- 10795440 TI - [Analysis of indications and diagnosis of the gastroscopies requested by primary care physicians]. AB - OBJECTIVES: To analyse the pathology diagnosed on the basis of the gastroscopies requested from a primary care centre (PCC). Assessment and treatment of Helicobacter pylori (HP) infection. DESIGN: Retrospective and descriptive. SETTING: Urban PCC. PATIENTS: Those over 14 who had a gastroscopy for a diagnostic purpose, requested by their primary care doctor during 1997. MEASUREMENTS: Review of clinical records and gastroscopy reports analysing: personal data, reasons for request, prior treatment, diagnoses, treatment to eradicate HP, later referral to the digestion specialist and waiting-time for the gastroscopy. RESULTS: 206 gastroscopies were performed, 139 for epigastralgia (67%) and 73 for heartburn (35%). 161 patients (78%) had had previous treatment, with alkalines used in 94 cases (45%). The most common diagnoses were: 54 normal (26%), 60 hiatus hernias (30%), 43 oesophagitis (21%), 18 duodenal ulcers (9%) and 4 gastric ulcers (2%). The Helicobacter pylori study was positive in 67 cases, with triple treatment (amoxycillin, clarithromycin and omeprazole) to eradicate it given for 7 days in 36 cases (54%). Referral to the specialist of 3% of the patients. Waiting-time for the performance of the gastroscopy: 60 days (SD: 35). CONCLUSIONS: Gastroscopy is a diagnostic method habitually used in our PCC. The pathology diagnosed is mainly treated in primary care. Given the current controversy about actions at determined clinical entities, standardisation of criteria used by professionals at different care levels is very important. This standardisation should affect the indications of the examinations, and the treatment and follow-up of the pathologies. The reduction in waiting-time for a gastroscopy could possibly modify the prescription and duration of the prior treatment. PMID- 10795441 TI - [Practical usefulness of genogram in the handicapped patient care]. AB - OBJECTIVE: To know the practical use of genogram in the handicapped patient care, describing their characteristics and those of their families and to obtain the prevalence of bad care immobilized patients belonging to the health centre. DESIGN: Descriptive observational. SETTING: Urban health centre. PARTICIPANTS: People of both sexes registered at the health centre, immobilized in their houses, with a disability score of three or more in the physical disability scale of the Red Cross. MEASUREMENTS AND RESULTS: The most important features of our immobilized people, obtained from the genogram, are: women (63.5%), with mean age of 78.8 +/- 13.2 years old (CI, 76.5-81.17), with disability score of 3 (46.8%), immersed in a nuclear family (34.9%), in phase VI of dissolution of the vital cycle (45.2%), with very closed relations (45.2%), with arthrosis (23.8%) as the most frequent immobilized cause, and with one unique home career only (72%). The prevalence of bad care immobilized patients was 25.4%. CONCLUSIONS: The genogram is an excellent tool for describing immobilized people and their family nucleus, allowing the definition of an immobilized profile. If the patient is bad cared, he will need an special consideration. The immobilized score and the family relations ship may complicate their care, conditioning a worse prognosis. We wish to emphasize the importance of carrying out the genogram in the initial moment of the immobilization, because it establishes a dialogue route between the families and the sanitary professionals to improve the attention of these patients. PMID- 10795442 TI - [Internet as a means of professional communication: electronic mailing lists]. PMID- 10795443 TI - [Primary care detection of alcohol consumption]. PMID- 10795444 TI - [Bioethics and family medicine (III). Work Group of the SemFYC]. PMID- 10795445 TI - [Spironolactone in heart failure]. PMID- 10795446 TI - [And spironolactone in the treatment of heart failure?]. PMID- 10795447 TI - [Decrease in mortality on spironolactone being added to the conventional treatment of heart failure]. PMID- 10795448 TI - ["Mononucleosis-like" caused by carbamazepine]. PMID- 10795449 TI - Myelointegration of titanium implants: B lymphopoiesis and hemopoietic cell proliferation in mouse bone marrow exposed to titanium implants. AB - Multinucleated giant cells have been observed at interfaces between bone marrow and titanium implants in mouse femurs. This raises concern that macrophage derived factors might perturb local lymphohemopoiesis, possibly even predisposing to neoplasia in the B lymphocyte lineage. It has been found that an implant marrow interface with associated giant cells persists for at least 1.5 years. Precursor B cells show early increases in number and proliferative activity. At later intervals, however, they do not differ significantly from controls, and there are no perturbations in spatial localization of either B lineage cells or DNA-synthesizing hemopoietic cells. The results of this investigation in mice demonstrate that, following initial marrow regeneration and fluctuating precursor B cell activity, and despite the presence of giant cells, titanium implants apparently become well-tolerated by directly apposed bone marrow cells in a lasting state of "myelointegration." PMID- 10795450 TI - Placement of implants in distraction osteogenesis: a pilot study in dogs. AB - This study investigated the possibility of achieving osseointegration of implants placed in a distracted site during the consolidation period. Four healthy male mongrel dogs were used in this experiment. A subperiosteal corticotomy around the mandible was performed between the left mandibular premolar and first molar. After a 7-day latency period for soft tissue healing, the distraction was performed at the rate of 1 mm per day for 14 consecutive days to allow for 14 mm of elongation, using an extraoral distraction device. Three weeks after the completion of distraction, screw-type implants were placed in the distracted site. Twenty-four weeks after placement of the implants, they were stable, and osseointegration had been achieved physically, radiographically, and histologically. These results suggest the possibility of shortening the period of implant treatment by using the distraction osteogenesis technique. PMID- 10795451 TI - Periodontal ligament formation around titanium implants using cultured periodontal ligament cells: a pilot study. AB - The aim of this study was to investigate whether a new periodontal ligament attachment will form on titanium implants when they are implanted with cultured periodontal ligament cells. Periodontal ligament cells obtained from the teeth of 3 dogs were cultured and attached to the surface of titanium implants. The implants with the cultured autologous periodontal ligament cells were placed in the mandibles of the dogs. After 3 months of healing, histologic examination revealed that, on some implant surfaces, a layer of cementum-like tissue with inserting collagen fibers had been achieved. These results demonstrated that cultured periodontal ligament cells can form tissue resembling a true periodontal ligament around implants. PMID- 10795452 TI - Assessment of osseointegration by nonlinear dynamic response. AB - Quantitative assessment of osseointegration remains a goal of researchers and clinicians alike. In this study, an instrument was designed for this purpose and tested in an animal model. Effective mechanical impedance, linearized for quasi static force, was measured in 22 implants placed in the hind tibiae of 2 large hounds. The results demonstrate that in this animal long-bone model, the effective impedance of titanium root-form implants exhibits a degree of nonlinear behavior correlated with their state of osseointegration. This observation may be the basis for useful clinical instrumentation. PMID- 10795453 TI - Analysis of stress distribution in a screw-retained implant prosthesis. AB - Four types of implant superstructures were screwed onto implant bodies, and the strains created around the implant bodies were compared and analyzed within the IMZ Implant System. Three IMZ implants were embedded in the center of a polyurethane block (30 x 40 x 30 mm), and a total of 16 superstructures was fabricated by 4 methods: 1-piece cast, 1-piece cast/split soldering, soldering, and passive fit. Six strain gauges were placed on the surface of the block 1 mm apart. Three embedded implants were numbered, and a fixed partial denture was placed on these implants and screwed by a torque wrench using 14.5 Ncm torque. This procedure was repeated 7 times for each fixed partial denture, and each created strain was measured when the last screw was tightened. In all fixed partial dentures, strains were produced around the implant bodies when screws retaining the prosthesis were tightened, and the strain was relieved with unscrewing. The magnitude of strain was greater with the 1-piece cast method or the section/solder method than with the soldering and passive-fit methods. Of the 2 soldering methods, when the screw on the middle implant was tightened before those on the terminal 2 implants, the magnitude of strain was lower with the soldering method than with the 1-piece cast/split soldering method. When the order of screw tightening was changed, there were significant differences in the magnitude of strain at each gauge with the soldering method. With the passive-fit method, no differences in the magnitude of strain attributable to the order of screw tightening could be detected. The magnitude of strain produced around a screw-retained implant prosthesis was significantly lower with the passive-fit method when compared to the other 3 fabricating methods. Furthermore, the implants prepared by the passive-fit method were not affected by the order of screw tightening. PMID- 10795454 TI - Cleaning and heat-treatment effects on unalloyed titanium implant surfaces. AB - This study tested the following hypotheses: (1) acid-cleaned and passivated unalloyed titanium implants have higher surface energies (which are considered desirable for bone implants) than ethanol-cleaned titanium; (2) higher temperatures of heat treatment of unalloyed titanium result in higher surface energies; and (3) these changes can be related to changes in surface composition and roughness. Thus, unalloyed titanium specimens were either acid-cleaned and passivated (CP) or ethanol-cleaned (Et). Each set was then divided into 3 groups and heat-treated for 1 hour at 316 degrees C (600 degrees F), 427 degrees C (800 degrees F), and 538 degrees C (1,000 degrees F), respectively. Surface roughness values for each of these groups were determined using atomic force microscopy, while surface compositions were determined using Auger electron, x-ray photoelectron, and Raman spectroscopic techniques. Surface energies were estimated using a 2-liquid geometric mean technique and correlated with surface roughness, elemental composition, and elemental thickness. The CP surfaces were slightly rougher than the Et specimens, which had greater oxide thickness and hydrocarbon presence. The surface oxides were composed of TiO2, Ti2O3, and possibly titanium peroxide; those heat-treated at 427 degrees C or above were crystalline. The CP specimens had carbonaceous coverage that was of a different composition from that on Et specimens. The CP specimens had significantly higher surface energies, which showed statistically significant correlations with oxide thickness and carbonaceous presence. In conclusion, ethanol cleaning of unalloyed titanium dental implants may not provide optimal surface properties when compared to cleaning with phosphoric acid followed by nitric acid passivation. PMID- 10795455 TI - Three-dimensional bone response to commercially pure titanium, hydroxyapatite, and calcium-ion-mixing titanium in rabbits. AB - Three-dimensional bone response to 3 biomaterials--commercially pure titanium (Ti), hydroxyapatite (HA), and calcium-ion-mixing titanium (Ca-Ti)--embedded in the tibiae of rabbits was examined chronologically. The rabbits were sacrificed at 2, 4, and 8 weeks after implantation, and the percent bone volume around each implant was calculated from the implant surface to each of 4 measurements: 36 microns, 0.25 mm, 0.5 mm, and 1.0 mm in 2 regions (cortical bone and bone marrow regions). Percent bone volume in the cortical bone was consistent, whereas in the bone marrow region, the percent bone volume varied according to implant material, implantation period, and distance from the implant surface. With Ti implants the percent increased gradually up to 8 weeks at each distance, whereas in HA and Ca Ti implants the percent was largest at 4 weeks and increased closer to the surface. The percent with Ti implants was largest at 36 microns to 0.25 mm. Aspect of bone response to Ca-Ti was its position intermediate between those of HA and Ti. The decrease of the percent at 8 weeks was smaller than HA. PMID- 10795456 TI - Passivity of fit and marginal opening in screw- or cement-retained implant fixed partial denture designs. AB - The relationship of stress generation upon placement of cement-retained or screw retained implant restorations has not been thoroughly investigated. Passivity of fit and marginal discrepancies of screw- and cement-retained implant fixed partial denture (FPD) designs were determined using a photoelastic model of a partially edentulous posterior mandibular arch with 3 screw-type implants. Buccal and lingual marginal openings, measured with a traveling microscope before cementation or screw tightening, revealed no statistical difference in adaptation between designs. Screw tightening caused a reduction in marginal opening (changes significant, P < .05). The opening with the cemented FPDs was similar before and after cementation. Photoelastic evaluation of the FPDs showed that cement retained FPDs exhibited a more equitable stress distribution than did their screw retained counterparts. PMID- 10795457 TI - Improvement of epidermal adhesion by surface modification of craniofacial abutments. AB - Craniofacial implants may present peri-implant inflammation because there is no close adhesion of the epithelium to abutments and because of bacteria infiltrating the subcutaneous tissue through the gap. Therefore an attempt was made to improve adhesion of epithelium to abutments. In an in vitro model, adhesion of epithelial cells (HaCat cells) to nonmodified and 3 modified Branemark System abutment surfaces was quantified. It was found that more cells were adherent in sequence at silicone-coated surfaces, sandblasted surfaces, and collagen-coated (Types I and IV) surfaces than on nonmodified abutments. It was concluded that it is possible to improve epidermal adhesion to abutments through modification of abutment surfaces. PMID- 10795458 TI - Adult growth, aging, and the single-tooth implant. AB - Single-tooth implants are an increasingly popular method for replacing single teeth. While the effects of growth on implants in children have been well documented, the changes that occur in adults have not been studied with respect to single-tooth implants. It has been assumed that adults are stable and do not change; however, research in the last few years has indicated that adults do change with aging, and adult growth does occur. The changes in adults occur over decades rather than rapidly, as seen in children. Aging changes are readily apparent in the soft tissues of the face and create dramatic changes. Changes in the jaws and teeth occur as a result of continued, slow growth, in contrast to the aging effects seen in soft tissues. Growth changes occur in the arches and result in adaptive changes in the teeth over time, both vertically and horizontally, and in alignment. These dental changes may result in a lack of occlusion vertically or malposition of adjacent natural teeth relative to the implant crown. Clinicians may be well advised to observe and report these changes and warn patients that changes can occur over the service life of the implant supported crown. These changes may require maintenance adjustments or possible remaking of the implant crown as a result of adult growth, wear, or the esthetic changes of aging. PMID- 10795459 TI - The challenge of endosseous implants placed in the posterior partially edentulous maxilla: a clinical report. AB - The survival rate of implants placed in the maxillary molar area in a 2-stage procedure was evaluated. Between 1990 and 1997, 60 consecutive patients (32 females and 28 males, mean age 51 years) received 87 implants to replace missing maxillary molar teeth. Radiographs were evaluated preoperatively for bone quantity (mesiodistal width, potential implant length not compromising the integrity of adjacent vital structures). Second-stage surgery was performed in a mean of 7.9 months postimplantation. The 5-year cumulative implant survival rate and the influence of implant characteristics (type, length, diameter, and coating) on implant failure and complication rates (between the 2 stages of surgery) were evaluated. The total 5-year cumulative survival rate was 95.4% (4 implants were lost). There were a total of 17 "complications" (premature spontaneous implant exposure) in non-failing implants, 11 with high and 6 with flat cover screws, respectively. Implantation in the edentulous maxillary molar area is a predictable procedure with a considerably high survival rate. The type of implant cover screw used can affect the complication rate. PMID- 10795460 TI - Tissue preservation and maintenance of optimum esthetics: a clinical report. AB - Today, most tooth replacement in the esthetic zone is done using implants placed in a delayed surgical protocol. Unfortunately, this delay can result in loss of both hard and soft tissue during the healing period, necessitating guided tissue regeneration techniques at the time of implant placement. Recent developments with tapered implants have facilitated predictable immediate implant placement, preserving the osseous structure surrounding the socket. Further developments with custom healing abutments can preserve the crestal soft tissues, including the papillae. This article reviews techniques that provide for the preservation of both bone and soft tissue while enhancing the esthetic results around implants. PMID- 10795462 TI - Fabrication of a dual-purpose surgical template for correct labiopalatal positioning of dental implants. AB - An implant-supported prosthesis can potentially provide a functional and esthetic reconstruction, provided that the implants are precisely placed in the predetermined location and angulation. Since the accurate placement of osseointegrated implants in the anterior maxilla with questionable bone support is often a clinical challenge, the use of computed tomography and surgical guides is crucial. This article describes a technique to fabricate a template that served as a guide for radiographic evaluation and was then modified for use in the surgical phase of the treatment. PMID- 10795461 TI - Evaluation of the precision obtained with a fixed surgical template in the placement of implants for rehabilitation of the completely edentulous maxilla: a clinical report. AB - The fabrication of a complete maxillary implant-supported prosthesis in a patient with slight resorption of the alveolar ridge and a high lip line presents a professional challenge. The implants must be placed with high precision to achieve good esthetics, phonetics, and function. A fixed surgical template using microimplants has been developed for this purpose. The objective of this investigation was to compare a fixed surgical template (FST) to a conventional movable surgical template (MST) for the precise placement of implants in the slightly resorbed edentulous maxilla. Three patients (28 implants), edentulous in the maxilla, with slight ridge resorption, in whom the implants were placed with an FST, were compared with 5 controls having the same characteristics and implants placed with an MST (35 implants). After completion of the prosthesis, occlusal photographs (1:1) were taken, and these images were scanned and transferred to a drawing program in which the contours of the teeth, the ideal emergence position of the occlusal hole of the abutment screw, and its real position were drawn. A blind evaluation was made using the following variables: frequency of location of the abutment screw hole outside of the tooth contour, and the relative measurements of the area of coincidence between the circle that represents the ideal position and real position. A significantly smaller frequency of implants outside the tooth contour was seen with the FST (7%) than with the MST (46%) (P < .0008). Also, a significantly higher relative area of coincidence was observed between ideal position and real position in the FST (0.61) than in the MST (0.38) (P < .003). This study revealed that considerably higher precision was associated with the use of an FST. PMID- 10795463 TI - Tissue reactions, fluids, and bacterial infiltration in implants retrieved at autopsy: a case report. AB - A 72-year-old patient underwent the placement of 2 screw-type implants. After 5 months the patient died of a massive stroke, and a block section of the portion of the mandible containing the implants was done. The specimen was treated to obtain thin ground sections. A 1- to 5-micron gap was present between the implant and the healing cover screw, and this space was filled by bacteria and calculus; bacteria were also present in the most apical portion of the hollow part of the implant. An inflammatory infiltrate was present in the connective peri-implant tissues. The spaces between all implant components (implant, abutment, and healing screw) can act as conduits and reservoirs for bacteria, which could cause inflammation of the peri-implant soft tissues. In conclusion, the histologic data from this autopsy case may help to confirm the penetration by fluids and bacteria into the internal portion of the implants, obtained from previous in vitro and in vivo studies. PMID- 10795465 TI - Pediatric cancers. PMID- 10795464 TI - Maxillary antral mucocele and its relevance for maxillary sinus augmentation grafting: a case report. AB - Paranasal sinus mucoceles are benign, locally expansile cyst-like masses that are filled with mucus and lined with epithelium. Most occur in the frontal sinus. Maxillary sinus mucoceles are presumably uncommon in the United States and European countries, although they have been frequently reported in Japan, particularly following Caldwell-Luc surgery. Clinical symptoms may not appear for at least 10 years postoperatively. Chronic sinus inflammation and allergic disease are also common causes of paranasal mucoceles. This paper provides an overview of maxillary sinus mucoceles and presents a case study involving a 62 year-old Latin male whose asymptomatic maxillary sinus mucocele was not revealed until he presented for maxillary sinus grafting and implant placement. PMID- 10795466 TI - Outcome of singleton term breech cases in the pretext of mode of delivery. AB - OBJECTIVE: A study was carried out to ascertain maternal and perinatal outcome of cases presented as breech versus their mode of delivery so that appropriate management strategy could be mapped out without compromising fetomaternal wellbeing. METHODS: The study was carried out in one of the units of the Department of Obstetrics and Gynaecology of Sir Ganga Ram Hospital, affiliated with Fatima Jinnah Medical College, from August, 1996 to July, 1999. Management of 352 singleton term breech cases was analyzed. Data gathered were socio demographic variables, detailed obstetrical history, important features of index pregnancy, type of breech presentation, selection of mode of delivery and reason for choosing a particular route of delivery, indications of elective or emergency caesarean sections in the cases applicable, fetal outcome and maternal complications. RESULTS: In 135 (38%) patients C-section (94 elective and 41 emergency) was done straightaway and trial of vaginal breech delivery was given to the remaining 217 (62%). Of these vaginal delivery could be done in 161 (74%) cases and 56 (26%) required caesarean section due to fetal distress and or failure to progress. About 32% primiparas could manage vaginally 55% multiparas while the ratio of caesareans done in both the groups was 68% vs 45%. A direct proportion was observed between C-section rate and neonatal birthweight. Route of delivery did not influence significantly neonatal outcome which become evident when no marked difference could be detected in corrected neonatal mortality rates calculated for the groups having elective C-section, emergency section and vaginal delivery. Perinatal mortality rate and corrected neonatal mortality rate of the study population and the same values computed for all the deliveries conducted during the study period were found to comparable. Maternal and neonatal complications were seen more frequently in the group requiring emergency caesarean section. CONCLUSION: Proper selection of cases for vaginal delivery, vigilant intrapartum monitoring and employing proper technique of breech delivery have been established as the most important determinants for successful vaginal breech delivery without compromising fetomaternal well being and curtailing the percentage of caesareans being done for this malpresentation. PMID- 10795467 TI - Frequency of malignant solid tumors in children. AB - OBJECTIVE: To find out the frequency of malignant solid tumors in children (< 15 years). SETTING: All cases of pediatric malignant solid tumors which were diagnosed in the section of histopathology at the Aga Khan University Hospital, Karachi during the period of two years. METHODS: These tumors were initially evaluated on H&E stained sections and special stains were also performed whenever indicated. The undifferentiated tumors were evaluated immunohistochemically by using a panel of antibodies on sections from routinely processed, formalin fixed, paraffin embedded tissue blocks. RESULTS: Of two hundred and fifty three (253) consecutive cases of paediatric malignant solid tumors, lymphoma (26.1%) was the most common tumor followed by central nervous system tumors (16.6%), osteosarcoma (7.5%), rhabdomyosarcoma (6.7%), neuroblastoma (5.1%), Wilm's tumor (5.1%), Ewing's sarcoma (4.7%), retinoblastoma (4.7%), germ cell tumor (4.4%) and primitive neuroectodermal tumor (4%) in order of frequency. In seven cases (2.8%), the nature of lesion remained undetermined even after immunohistochemical staining. Rest of malignant tumors (12.3%) included the rare entities like synovial sarcoma, nasopharyngeal carcinoma, leiomyosarcoma, malignant schwannoma and thyroid carcinoma, etc. CONCLUSION: Lymphoma was the most frequent Paediatric tumor. The frequency of childhood central nervous system tumors was quite high as compared to the other series from different regions of Pakistan. PMID- 10795468 TI - Childhood non-Hodgkin's lymphoma: an immunophenotypic analysis. AB - OBJECTIVE: To observe the frequency of histological subtypes of childhood non Hodgkin's lymphoma and its immunohistochemical profile. SETTING: All cases of non Hodgkin's lymphoma diagnosed in children (< 15 years) in the section of histopathology at the Aga Khan University Hospital Karachi during the period of three years. METHODS: These consecutive cases were evaluated on H&E stained sections and then immunohistochemistry analysis of these tumors was performed by employing Peroxidase Anti-Peroxidase (PAP) technique. RESULTS: The present series included 61 cases of non-Hodgkin's lymphoma. NHL was more common in males as compared to females with male to female ratio of 5.8:1. High grade NHL comprised 87% of childhood lymphoma. The mode of presentation in majority of NHL (57%) was extranodal. Burkitt's lymphoma (33%) was the most prevalent histological subtypes, followed by lymphoblastic (28%), diffuse large cell (15%), diffuse mix small and large cell (13%), small non cleaved Non-Burkitt's (7%) and immunoblastic (4%). Immunophenotypic analysis of the childhood Non-Hodgkin's lymphoma revealed that 67% of the Non-Hodgkin's lymphoma are B-cell type while 33%, are those of T-cell lineage. CONCLUSION: NHL was more common in males. Majority of NHL in children were high grade tumors. Burkitt's lymphoma was the most frequent histological subtype. T-cell NHL comprised a significant portion of childhood lymphomas. PMID- 10795469 TI - Neonatal sepsis: an etiological study. AB - OBJECTIVE: A periodic review of neonatal sepsis to asses any change in the infecting organism. METHOD: A prospective study was conducted at HMC and ASH, Karachi. The babies suspected to have or developed sepsis any time during hospitalization were investigated to establish the diagnosis and isolate the causative organism. Blood culture was taken at the time of admission or when sepsis was suspected. RESULTS: Out of 109 episodes of blood culture proven sepsis 68 presented as early onset (within 48 hours of birth) and 41 as late onset sepsis (after 48 hours of birth). In early onset group Gram -ve and Gram +ve organisms were almost equal, i.e. 33 and 35 respectively. Among the gram -ve organism most of the cases were due to Klebsiella sp, and Enterococcus was the commonest Gram +ve organism. In late onset group majority of infections were due to gram +ve organisms, i.e. 30 out of 41. Staph. aureus and Staph. epidermidis were commonest. The organisms were least sensitive to Ampicillin (< 20%) and highly sensitive to Amikacin (90% to 100%), Cefotaxime was also seen as a good choice of antibiotic with sensitivity of (84%-89%). CONCLUSION: Gram +ve organisms were the main cause of neonatal sepsis. Klebsiella sp. is still the commonest organism causing early onset sepsis. The data must be periodically reviewed and antibiotic policy revised accordingly. PMID- 10795470 TI - Rapid identification of neonatal sepsis. AB - OBJECTIVE: To achieve rapid identification of neonatal sepsis. SETTING: Neonatal Intensive care unit (NICU) of a teaching hospital. METHOD: We evaluated fifty neonates who were admitted with clinical features suggesting sepsis or who had principal risk factors, e.g. Prematurity (< 36 weeks), Low birth weight (< 2.5 kg), H/o maternal pyrexia or prolonged rupture of membranes, birth asphyxia, unbooked cases or instrumentation. Five tests, i.e., Total Leukocyte Count (T.L.C.), Absolute Neutrophil Count, Immature/Total Neutrophil ratio (I.T. ratio), Platelet count and C-Reactive protein were used for rapid diagnosis of neonatal sepsis. RESULTS: C-reactive protein (C.R.P.) and absolute Neutrophil count had a sensitivity of over 60% with a specificity of 50%. White blood cell count had a specificity of 93% but a sensitivity of 14%. CONCLUSION: None of the tests used alone were reliable, but when in combination these five tests may help to diagnose sepsis within a few hours. Also, if the tests show a high negative predictive value, the neonate can be discharged early from the hospital, stopping the antibiotics, thereby reducing the cost of treatment and anxiety of the family. PMID- 10795471 TI - Levels of uric acid, urea and creatinine in Iraqi children with sickle cell disease. AB - OBJECTIVE: To determine the levels of serum uric acid, urea and creatinine in subjects with sickle cell disease and compare them to those reported in literature. SETTING: Department of Paediatrics, College of Medicine, University of Basrah. METHODS: Plasma uric acid, urea and creatinine was estimated by Varley's method, group of 65 sickle cell patients (35 Hb AS, 30 Hb SS) aged between 2-11 years. The results were compared with those obtained in a group of 45 age and sex-matched controls with normal haemoglobin (Hb AA). RESULTS: The uric acid level was elevated in sickle cell patients as compared with the normal control group. The 95% confidence intervals for differences in the mean of the two groups: HbAA vs HbAS was 4.22 (0.3), while for HbAA for HbSS was 3.4 (0.06), both being statistically highly significant [p < 0.0001]. Urea and creatinine levels were considerably lower in the sickle cell disease patients. The difference in the patient's mean for urea compared to the mean in the normal group (Hb AA) was 9.64 (1.95) and 8.55 (1.76) for HbSS and HbAS, respectively. Like wise, the difference in the mean for creatinine in HbSS group was 0.71 (0.12) and in HbAS was 0.76 (0.12), which was statistically significant [p < 0.0001]. CONCLUSION: Raised serum uric acid levels were found in Iraqi children with sickle cell disease, creatinine clearance studies will be valuable to assess renal function. PMID- 10795472 TI - Resuscitative fluids in penetrating trauma: when and when not to give? PMID- 10795473 TI - Endovascular embolization of traumatic intracavernous pseudoaneurysm of internal carotid artery. PMID- 10795474 TI - Post-graduate medical training in Singapore. PMID- 10795475 TI - [Creative thinking in chronic cough. The etiology could be in the ear...]. PMID- 10795476 TI - [Unconventional cancer therapy on trial. Mistletoe preparations: only effective in the test tube?. Interview by Elke Runge]. PMID- 10795477 TI - [The patient is always tired. Testing for hepatitis C?]. PMID- 10795478 TI - [Erythrocytes in the urine: glomerulonephritis or other source of bleeding. Answer with the microscope]. AB - On the basis of an evaluation of the morphology of erythrocytes in urinary sediment, it is possible to distinguish between nephrological and urological hematuria. In the case of the former, the cells have "disruptive characteristics" (membrane evagination, ring formations, dwarf forms and loss of hemoglobin), enabling them to be distinguished from the regularly formed erythrocytes from hemorrhage within the urinary tract. Dysmorphic erythrocytes retain their characteristic shapes even when persisting in the urine for lengthy periods. An assessment of erythrocyte morphology forms a suitable basis for choosing appropriate further diagnostic measures for urological or nephrological work-up. The method of examination is simple and readily reproducible, and should therefore form an early part of the diagnostic strategy. Microscopic assessment erythrocyte shape is not suitable for the diagnostic work-up of tumors, nor can it obviate the need for kidney biopsy for the investigation of histomorphological changes in glomerular nephritis. PMID- 10795479 TI - [Nephrologic emergency or bland course? Why glomerulonephritis should be checked out]. AB - Glomerulonephritis frequently takes an asymptomatic course, and may lead on to chronic renal insufficiency. If clinical signs of glomerulonephritis are present, the syndrome must be characterized by means of a comprehensive diagnostic work up. The definitive diagnosis can be established only on the basis of a renal biopsy, which also permits an assessment of the prognosis. In this review, the basic pathogenetic principles are described in brief. Taking the most important forms of glomerulonephritis--IgA nephropathy, membranous glomerulonephritis and rapidly progressive glomerulonephritis--as examples, the general therapeutic options and a number of specific treatment strategies are discussed. New facts relating to the mechanisms of glomerular damage will in future make possible the use of targeted therapeutic interventions. PMID- 10795480 TI - [Somatisizing patients. Concrete recommendations for a stepwise therapy in general practice]. AB - Patients with physical symptoms but no detectable organic substrate often present a challenge for the family doctor. The therapeutic approach discussed herein focuses on the development of a psychosomatic model of the problem. In this step by-step approach, the patient's subjective understanding of, and attitude towards, his illness and its treatment are established, alternative psychosomatic models of disease based on a consideration of psychophysiological interactions are worked out jointly with the patient, and, finally, thought is given to the current complaints within the context of the patient's personality and life situation. The basis for successful treatment is an empathic and sound doctor patient relationship based on trust that gives the patient the feeling that he and his problem are being taken seriously. The prerequisite for the use of such a program is appropriate training in basic psychosomatic care. PMID- 10795481 TI - [Oral antidiabetic drugs--1. For each patient the proper drug]. PMID- 10795482 TI - [Phytotherapeutic drugs for the common cold. Results of a randomized controlled double-blind study]. PMID- 10795483 TI - [Ulcus cruris. 2: Therapy]. PMID- 10795484 TI - [Diagnostic quiz. Diarrhea and muscle weakness. Whipple disease]. PMID- 10795485 TI - [No liquidity analysis in the program? Then your tax counselor is not effective]. PMID- 10795486 TI - [The patient and the informed consent]. PMID- 10795488 TI - [Magnetic resonance tomography]. PMID- 10795487 TI - [Atopic diseases]. PMID- 10795489 TI - [When everybody does his job, health services will improve]. PMID- 10795490 TI - [Clinical neurological examination of patients referred to a neurological department]. AB - BACKGROUND: We studied reports of clinical neurological status in patients referred from primary and secondary health service to a neurological department. MATERIAL AND METHOD: We evaluated referrals consecutively during 6 months. By using parametric and categorical statistical methods, we studied characteristics in examined patients compared with others. RESULTS: Of a total of 912 patients, 431 (47%) had been examined neurologically. Among the women, 267 women (51%) had been examined; among the 164 men (43%). Patients with musculoskeletal disorders were more often examined, p = 0.0001, while those with faint or seizure disorders were significantly less frequently examined compared with other patients, p = 0.009. Clinical neurological examinations were performed more often in women compared with men, p = 0.03. The mean age was higher in the examined group (49.7 years) compared with others (46.5 years), p = 0.006. INTERPRETATION: Clinical neurological examination in patients referred to a neurological department had more often been performed in women, in higher age groups, and in those with musculoskeletal symptoms. In more than half of the patients, the decision of referring patients to the department of neurology was not based on results obtained from clinical neurological examination. PMID- 10795491 TI - [Tongue atrophy--a marker of malnutrition]. AB - BACKGROUND: Atrophic glossitis is considered to be a marker for nutritional deficiency. Study of the tongue is part of the clinical examination. The importance of atrophic tongue has been unclear. As part of a bigger nutritional investigation in the elderly, we wanted to study the occurrence of atrophic tongue and relate this finding to other nutritional measures. MATERIAL: The presence of atrophic tongue was studied in 311 hospitalized, and in 106 randomly selected elderly at home. RESULTS: Atrophic tongue was present in 100 of 311 hospitalized patients, and only in ten out of the 106 persons living in their own homes. Compared to those without, patients with atrophic tongue had significantly lower weight, body-mass index, triceps skinfold, arm muscle circumference and increased Katz index values for independence in daily activities. Patients with atrophic tongue had significantly reduced concentrations of serum cholesterol, ascorbic acid and calcidiol. Tongue atrophy had a sensitivity for serious malnutrition 0.30, for serious and moderate malnutrition together 0.70. The specificity for malnutrition was 0.76. INTERPRETATION: We conclude that atrophic tongue is common in elderly people and a marker for malnutrition and reduced muscle function. PMID- 10795492 TI - [Dementia and hypothyroidism]. AB - BACKGROUND: The clinical presentation of hypothyroidism varies considerably and may be dominated by cognitive deficits and psychological symptoms. Hence, when diagnosing dementia it is important to rule out hypothyroidism. Hypothyroidism has also been considered a classic reversible dementia; however, the aetiological association between dementia and hypothyroidism is by no means indisputable. MATERIAL AND METHODS: A review of available literature concerning hypothyroidism, cognitive functioning and dementia. RESULTS: The importance of thyroxin for the development of the brain and for intellectual development is well known. Early reports suggested that thyroxin improved or reversed dementia in patients with hypothyroidism. Later studies have shown contradictory results; however, several prospective intervention and follow-up studies of overt and subclinical hypothyroidism do not provide for support the belief that there is an aetiological association between dementia and hypothyroidism. INTERPRETATION: The lack of association between dementia and hypothyroidism has consequences for the treatment, and for information to patients and relatives of patients with concomitant dementia symptoms and hypothyroidism. The introduction of acetylcholinesterase inhibitors in the treatment of patients with Alzheimer's disease has brought new interest to this matter. When hypothyroidism is detected in a patient with suspected dementia, thyroxin should be tried first of all. If there is no obvious improvement of symptoms and there still is evidence of dementia, one should consider whether the patient is suitable for a trial with an acetylcholine inhibitor. PMID- 10795493 TI - [Cost effective and quality assured (adeno)tonsillectomy in children]. AB - BACKGROUND: The issues of whether minor surgery should be performed in private or public clinics and/or be funded by the national health service are under continuous debate in Norway. MATERIAL AND METHODS: A prospective study of the benefit of (adeno)tonsillectomies in 120 children is presented. Surgery included in the study was performed on an inpatient as well as an outpatient basis and in public as well as private clinics. RESULTS: There was no difference in patient satisfaction or in the quality of surgery depending on the type of organisation. Relief of symptoms associated with (adeno)tonsillar infections and obstruction following surgery was consistent. Less euresis was found in the study population following surgery. Increased haemoglobin concentration and less protoporfyrin IX in erythrocytes indicate an improved erythropoiesis following (adeno)tonsillectomy. Less manpower was needed to perform (adeno)tonsillectomies in private compared to public clinics. INTERPRETATION: We suggest that the Norwegian national health system opens up for funding surgery of established quality irrespective of whether it is performed in a public or a private clinic. PMID- 10795494 TI - [Withdrawal syndrome after the use of serotonin reuptake inhibitors]. AB - BACKGROUND: The use of serotonin reuptake inhibitors in the treatment of depression has increased significantly over the years since their introduction in Norway in the early 1990s. Reports of side effects have paralleled the increased use; one side effect is the withdrawal syndrome associated with serotonin reuptake inhibitors. MATERIAL AND METHODS: This article reports on two patients who discontinued paroxetine and experienced unusual shock-like symptoms consistent with this syndrome. RESULTS: There are numerous reports documenting withdrawal reactions caused by these antidepressants. Recently the first controlled studies have been published. INTERPRETATION: This syndrome is not well known and differs from the withdrawal syndrome caused by tricyclic antidepressants. PMID- 10795495 TI - [Does reduced microbial exposure contribute to increased prevalence of allergy?]. AB - Diseases associated with atopic allergy such as asthma, hay fever and eczema have increased dramatically in westernised societies over the last decades. Reduced microbial exposure during the first years of life, which we find in societies with a high standard of living, may be disadvantageous in terms of mucosal sensitisation to allergy. Recent studies have suggested an inverse relationship between allergic disorders and infections with intracellular pathogens including tuberculosis, measles, and hepatitis A. Because many intracellular pathogens generate predominantly a T helper 1 (Th1)-cell cytokine profile, and because Th1 and Th2 cell-mediated immune mechanisms are subjected to cross-regulation, it is possible that exposure to certain microorganisms may repress or abolish development of atopic allergy. PMID- 10795496 TI - [Does postmenopausal hormone replacement therapy have a place in treatment of hyperlipidemia?]. AB - Postmenopausal women experience increases in total cholesterol, triglyceride and lipoprotein (a) levels, decreased HDL cholesterol level and a shift in the LDL particle distribution toward a more dense subtype. Postmenopausal hormone substitution lowers LDL cholesterol level by 12-13%, also in women with hyperlipidaemia, and lowers lipoprotein (a) level. The effect on subtype of LDL has been variable in studies to date. The type, dose and mode of administration of the gestagen as well as the choice of oestrogen (conjugated equine oestrogen versus oestradiol) may affect triglyceride and/or HDL cholesterol levels. The first randomised study of postmenopausal hormone substitution in women with cardiovascular disease (Heart and Estrogen/progestin Replacement Study) showed an increase in cardiovascular events during the first year of therapy with a decrease by the fourth year, thus, there was no overall effect. Several new studies including the Women's Health Initiative and the Women's International Study of long Duration Oestrogen after Menopause should have the power to show the effect of postmenopausal hormone substitution on cardiovascular endpoints in primary prevention, however, the results of these studies will not be available for several years. PMID- 10795497 TI - [Is thrombolytic therapy effective in stroke?]. AB - BACKGROUND: The Norwegian Centre for Health Technology Assessment was asked to assess the treatment of stroke by trombolytic medication. We were also asked to include an evaluation of potential consequences for the organisation of the health care system. MATERIAL AND METHODS: Relevant literature was identified on Medline, the Cochrane Library (Systematic Reviews), and INAHTA (Systematic Reviews). Of particular importance are one study from the USA and two studies originating in Europe. RESULTS: The conclusion of the working group is that the benefit of thrombolytic treatment in the studies assessed is uncertain; furthermore, intracranial haemorrhage can occur as a complication of the treatment. INTERPRETATION: Future thrombolytic treatment of stroke patients should be carried out according to a research protocol with thorough documentation of effects as well as side effects. PMID- 10795499 TI - [Magnetic resonance angiography]. AB - INTRODUCTION: MR angiography (MRA) is a group of MRI techniques capable of producing projectional images of arteries and veins without the use of ionising radiation or catheters. MATERIAL AND METHODS: This review describes two of the main techniques, time-of-flight (TOF) MRA and phase contrast MRA, that provide high signal-intensity vessels due to flow phenomena, and a third main technique, contrast-enhanced MRA, which relies on intravenous injection of contrast media. Contrary to the iodine-containing x-ray contrast media, the contrast media for MRA are not nephrotoxic, making contrast-enhanced MRA a good alternative for patients with reduced renal function. MRA is indicated in several vascular areas, especially head and neck arteries and veins, the large thoracic and abdominal arteries and veins, and the lower extremity arteries. INTERPRETATION: For several indications, the sensitivity and specificity of MRA is approaching those of the gold standard, x-ray angiography. There are reasons to believe that most diagnostic angiographic procedures will soon be performed as MRA. PMID- 10795498 TI - [Magnetic resonance--historical and theoretical basis]. AB - Magnetic resonance (MR) imaging is a fast developing modality that has become an indispensable diagnostic tool at numerous disease states. The first MR imager in Norway was installed in 1986; by the end of 1999 Norway will have a total of 39 units, approximately one unit per 114,000 inhabitants. MR technique uses radiowaves and magnetic fields, and no ionising radiation is involved. Imaging is based on the fact that all biological tissues are magnetized when placed in a strong, static magnetic field. Short radiofrequency pulses set the tissue magnetic vector into rotation, and the vector induces electric currents in a receiver coil. The electric signals are spatially encoded by means of magnetic field gradients, thus enabling image reconstruction by means of Fourier transformation. The signal intensities can be made dependent on several tissue parameters, thus creating several unique image contrast possibilities. PMID- 10795500 TI - [Low use of radiotherapy in Norway--a scientific theoretical perspective]. PMID- 10795501 TI - [The change from clozapine to olanzapine--same effect?]. PMID- 10795502 TI - [Ozone in connection with ultraviolet rays disinfection]. PMID- 10795503 TI - [Sleep disorders]. PMID- 10795504 TI - [Alternately on alternatives]. PMID- 10795505 TI - [What is the most important in the courses on pulmonary rehabilitation?]. PMID- 10795506 TI - [Different guidelines are dangerous for continuing education]. PMID- 10795507 TI - Society of Cardiovascular Anesthesiologists 22nd annual meeting. Orlando, Florida, USA. May 6-10, 2000. Abstracts. PMID- 10795508 TI - Society for Obstetric Anesthesia and Perinatology 32nd annual meeting. Montreal, Quebec, Canada. May 31-June 4, 2000. Abstracts. PMID- 10795509 TI - The British Society of Gastroenterology annual meeting. Birmingham, 21-23 March 2000. Abstracts. PMID- 10795511 TI - 24th National Cancer Congress of the German Cancer Society. Berlin, Germany, 20 23 March 2000. Abstracts. PMID- 10795510 TI - Association for Research in Vision and Ophthalmology annual meeting. Fort Lauderdale, Florida, USA. April 30-May 5, 2000. Abstracts. PMID- 10795512 TI - Pathological Society of Great Britain and Ireland 180th meeting. London, United Kingdom, 18-21 January 2000. Abstracts. PMID- 10795513 TI - American Academy of Neurology 52nd annual meeting. San Diego, California, USA. April 29-May 6, 2000. Abstracts. PMID- 10795514 TI - Molecular epidemiology of outbreaks of gastroenteritis associated with small round structured viruses in Germany in 1997/98. AB - The molecular epidemiology of small round structured viruses (SRSVs) in Germany was studied using fecal specimens from 16 SRSV-associated gastroenteritis outbreaks in different parts of Germany during 1997/1998 by reverse transcription polymerase chain reaction (RT-PCR) and by sequencing of the ORF1 and ORF3 amplicons. The majority of the isolates clustered in one subtype and were closely related to published SRSV sequences of genogroup II. PMID- 10795515 TI - Selective HIV-1-induced downmodulation of CD4 and coreceptors. AB - CD4 and members of the chemokine receptor family are required for infection of host cells, in vitro and in vivo, by the human immunodeficiency virus type-1. Although it is established that HIV-1 gp 120 interacts with CD4 and the coreceptors CCR5 or CXCR4 at the plasma membrane during HIV entry, longer-term interactions taking place between these molecules and HIV Env are less well understood. We have measured the cell surface expression of CD4, CCR5 and CXCR4 on a CD4+/CXCR4+CCR5+ T cell line following infection by cell line-adapted X4 and primary X4, X4R5 and R5 viruses. We report a selective downmodulation of CD4 by X4 and R5X4 viruses, but not by R5 viruses. None of the viruses tested significantly reduced CXCR4 expression at any time after infection. CCR5 protein and mRNA expression was eliminated by chronic infection with R5 viruses. These results indicate that chronic HIV-1 infection has distinct effects on CD4 and coreceptor membrane expression that depends on viral origin and coreceptor usage. PMID- 10795516 TI - Improvement of a serodiagnostic strategy for foot-and-mouth disease virus surveillance in cattle under systematic vaccination: a combined system of an indirect ELISA-3ABC with an enzyme-linked immunoelectrotransfer blot assay. AB - Foot-and-mouth disease (FMD) recombinant non-capsideal viral antigens 3A, 3B, 2C, 3D and 3ABC were assessed individually in an indirect enzyme-linked immunosorbent assay (I-ELISA) for their ability to screen for persistent infection-specific antibodies in cattle, regardless of vaccination condition. Results of sequential serum samples from non-vaccinated animals with experimentally induced persistent infection, and their correlation with virus isolation, indicated that the polypeptides 3A, 3B and 3ABC showed the most adequate characteristics for further field studies. Reliable performance of the I-ELISA with the selected antigen 3ABC was indicated by the distinct patterns observed for the frequency distribution values of naive and true positive samples. For regularly vaccinated livestock, a clear negative profile was proved in samples from regions without recent history of FMD. In contrast, at 90 and 900 days post-outbreak, coexistence of a positive and a negative population was established. These findings indicated that, irrespective of vaccination, the test allowed a classification of the herd disease status. A high degree of agreement was observed between I-ELISA-3ABC and EITB results for clearly reactive and non-reactive sera. For samples with reactivity values close to that of the cut-off, the EITB profiles upheld the definition of the infection condition. On this basis, screening by I-ELISA-3ABC, together with confirmation of suspect or positive samples by EITB is proposed as an adequate and accurate approach for large-scale epidemiological surveillance. PMID- 10795517 TI - Inhibitory activity of the human papillomavirus type 1 AU-rich element correlates inversely with the levels of the elav-like HuR protein in the cell cytoplasm. AB - We have analysed the activity of the negative element in the late 3' untranslated region of the cutaneous human papillomavirus type 1 (HPV-1) mRNAs in a panel of epithelial cells, of both mucosal and cutaneous origin. The inhibitory activity was cell line dependent. Determination of the levels of heterogeneous ribonucleoprotein C1/C2 and HuR that have been shown previously to interact specifically with the HPV-1 negative RNA element revealed that low levels of the HuR protein in the cytoplasmic fraction correlated with high inhibitory activity of the HPV-1 element, whereas cell lines displaying low inhibitory activity contained high levels of HuR in the cytoplasm. These results suggested that the HuR protein has a positive effect on HPV-1 late gene expression. PMID- 10795518 TI - Molecular epidemiology of herpes simplex virus type 1 genital infection in association with clinical manifestations. AB - The antigenic types of herpes simplex virus (HSV), HSV-1 and HSV-2 are considered to be the etiology of genital herpes. Symptoms of primary HSV-1 and HSV-2 genital infections are similar, however, recurrence of the infection is less frequent after the HSV-1-related genital infection. We determined genotypes of 79 HSV-1 strains isolated from genital lesions in women (43 from primary and 36 from recurrent infections), by analyzing restriction fragment length polymorphism of the HSV-1 strains. Each proportion of genotypes of F1, F12, and F41 in strains isolated from recurrent genital lesions was higher than the corresponding proportion in strains from primary genital lesions. Genotypes of HSV-1 strains isolated two or more times from recurrent genital lesions of each of three subjects were genotype F1, thereby supporting the hypothesis that the F1 genotype is closely associated with recurrence. While the possibility of a genotype preference at the site of infection was not ruled out, genotypes of more than half the number of HSV-1 strains from genital lesions were the same as those from non-genital (mainly oral-facial) lesions analyzed in our foregoing studies, thus indicating that most HSV-1 genotypes are apparently shared by genital and non genital lesions. PMID- 10795519 TI - Definition of an epitope on Japanese encephalitis virus (JEV) envelope protein recognized by JEV-specific murine CD8+ cytotoxic T lymphocytes. AB - We defined an epitope on the Japanese encephalitis virus (JEV) envelope (E) protein recognized by CD8+ cytotoxic T lymphocytes (CTLs). CTLs induced in JEV infected BALB/c (H-2d) mice recognized E and/or premembrane (PrM) proteins, while CTLs in C57BL/6J (H-2b) and C3H/HeJ (H-2k) mice did not. JEV-specific CTLs had a phenotype of CD3+ CD4- CD8+. Twenty-four 9-amino acid (a.a.) peptides, which had binding motifs for H-2Kd, H-2Ld or H-2Dd, were synthesized according to the amino acid sequences of PrM and E proteins. CTLs induced by JEV infection recognized only the peptide K-3. Immunization of BALB/c mice with only a group of peptides including K-3 induced CTLs which recognized the homologous K-3 peptide, while immunization with other peptides did not. The peptide K-3 had a binding motif for H-2Kd. This is consistent with the finding that JEV-specific CTLs in BALB/c mice was H-2Kd-restricted. These results indicate that the epitope recognized by CTLs in BALB/c mice is located between a.a. 60 and 68 on the E protein, corresponding to an a.a. sequence of CYHASVTDI. PMID- 10795520 TI - High frequency of postnatal transmission of TT virus in infancy. AB - DNA of TT virus (TTV), a novel human circovirus, was tested for in 116 mother infant pairs who had participated in the adult T-cell leukemia prevention program (APP) in Nagasaki, Japan, and refrained from breast-feeding. By polymerase chain reaction with Okamoto's seminested primers, 36 of the 115 (31%) mothers were positive. At the age of 6-8 months, 7 of 29 (24%) and 6 of 72 (8%) infants born to infected and uninfected mothers were positive, respectively (P = 0.047; RR, 2.90). Maternal TTV DNA load did not correlate with infantile infections. Since 99 of 100 (99%) cord blood samples were negative and all the mothers refrained from breast-feeding, the infantile TTV transmission would not be intrauterine or milk-borne. Between 6-8 and 12-21 months of age, 4 of 12 (33%) and 5 of 22 (23%) children born to infected and uninfected mothers turned positive, respectively (NS). At 12-21 months of age, 8 of 21 (38%) and 12 of 32 (38%) children born to infected and uninfected mothers were positive, respectively (NS). These results indicate that the TTV infection prevails in children at a frequency comparative to that in their mothers within the first 2 years of life, regardless of the maternal TTV status. PMID- 10795521 TI - Genetic variability of measles viruses circulating in the Benelux. AB - In Europe measles incidence remains high and in some parts the disease is likely to be still endemic due to insufficient vaccination. Luxembourg experienced an outbreak with at least 110 cases in 1996, and cases continued to be reported throughout 1997. We used molecular epidemiology to investigate this apparent endemicity. On the basis of their N gene sequences, the isolates were assigned to the typical European C2 and D6 genotypes. Sequence diversity within the outbreak was 0.2%. The nucleotide distance between the C2-viruses of the outbreak and the other C2 isolates was at least three or four times higher, suggesting an independent origin of the latter viruses. Similarly, the four D6 viruses found in Luxembourg were thought to be of at least two or three origins. Thus, we propose here to use intra-outbreak sequence diversity to differentiate between sporadic endemic cases and a "pseudo-outbreak" of multiple unrelated imported cases. PMID- 10795522 TI - Grapevine fleck virus-like viruses in Vitis. AB - Two sets of degenerate primers for the specific amplification of 572-575 nt and 386 nt segments of the methyltransferase and RNA- dependent RNA polymerase cistrons of members of the genera Tymovirus and Marafivirus and of the unassigned virus Grapevine fleck virus (GFkV) were designed on the basis of available sequences. These primers were used for amplifying and subsequent cloning and sequencing part of the open reading frame 1 of the genome of GFkV, Grapevine asteroid mosaic-associated virus (GAMaV) and of another previously unreported virus, for which the name Grapevine red globe virus (GRGV) is proposed. Computer assisted analysis of the amplified genome portions showed that the three grapevine viruses are phylogenetically related with one another and with sequenced tymoviruses and marafiviruses. The relationships with tymoviruses was confirmed by the type of ultrastructural modifications induced in the host cells. RdRp-specific degenerate primers were successfully used for the aspecific detection of the three viruses in crude grapevine sap extracts. Specific virus identification was obtained with RT-PCR using antisense virus-specific primers. PMID- 10795523 TI - Comparisons of coat protein gene sequences show that East African isolates of Sweet potato feathery mottle virus form a genetically distinct group. AB - Sweet potato feathery mottle virus (SPFMV, genus Potyvirus) infects sweet potatoes (Ipomoea batatas) worldwide, but no sequence data on isolates from Africa are available. Coat protein (CP) gene sequences from eight East African isolates from Madagascar and different districts of Uganda (the second biggest sweet potato producer in the world) and two West African isolates from Nigeria and Niger were determined. They were compared by phylogenetic analysis with the previously reported sequences of ten SPFMV isolates from other continents. The East African SPFMV isolates formed a distinct cluster, whereas the other isolates were not clustered according to geographic origin. These data indicate that East African isolates of SPFMV form a genetically unique group. PMID- 10795524 TI - Herpes simplex virus type 2 infection of macrophages impairs IL-4-mediated inhibition of NO production through TNF-alpha-induced activation of NF-kappaB. AB - Nitric oxide (NO) production in macrophages by the interferon (IFN)-gamma inducible NO synthase has been shown to play a role in clearance of viral infections. We have previously shown that IFN-gamma-induced NO production is augmented by herpes simplex virus type 2 (HSV-2) infection through autocrine tumour necrosis factor (TNF)-alpha secretion and is inhibited by interleukin (IL) 4. Here we investigated the effect of HSV-2 infection on the inhibitory function of IL-4. Virus infection of mouse J774A.1 macrophages strongly reduced the ability of IL-4 to inhibit IFN-gamma-induced NO production, even at very high IL 4 concentrations. The effect of HSV-2 infection did not involve the IL-4 signal transduction pathway through STAT6. IL-4 reduced virus-induced TNF-alpha secretion and nuclear factor (NF)-kappaB activation significantly, but less in cells concomitantly treated with IFN-gamma. Furthermore, neutralisation of residual TNF-alpha activity or inhibition of NF-kappaB activation largely restored the inhibitory effect of IL-4. The data show that inhibition of IFN gamma-induced NO production by IL-4 is impaired by HSV-2 infection due to autocrine TNF-alpha-mediated NF-kappaB activation. We suggest that the described phenomenon might be beneficial for the host by limiting high and sustained NO production to infectious foci. PMID- 10795525 TI - Sequence variability in the coat protein gene of two groups of banana bunchy top isolates. AB - Complete nucleotide sequences of the coat protein gene (DNA-3) of banana bunchy top virus (BBTV) were obtained from five geographical isolates by PCR. Analysis of these sequences revealed two distinct groups of BBTV isolates with those from the Philippines, Taiwan and Vietnam forming the Asian group while the South Pacific/African group consisted of isolates from Australia, Burundi and Fiji. At the nucleotide level, the sequences of DNA-3 were more similar between isolates from the same group (maximum 5.86%) than between members of the two different groups (maximum 13.05%). At the amino acid level, the BBTV coat protein remained highly conserved, with a maximum of < 3% sequence variation between all isolates in this study. There was a significantly higher degree of divergence between the Asian isolates, which may indicate that BBTV has been present in this region for an extended period of time or that there have been multiple introductions of BBTV into bananas. The high level of conservation in the BBTV coat protein suggests that any of the DNA-3 sequences presented in this study would probably be equally effective as transgene in attempts to generate transgenic banana plants with resistance to both groups of BBTV isolates. PMID- 10795526 TI - High prevalence of serum antibodies to Ras and type 16 E4 proteins of human papillomavirus in patients with precancerous lesions of the uterine cervix. AB - Serum samples from 38 healthy women and 55 women with different types of cervical lesions were investigated for the presence of antibodies to Ras and against E4 and E7 proteins of human papillomavirus type 16 (HPV-16). Our results showed that anti-E7 antibodies were closely associated with cervical cancer (75%), as previously reported. Interestingly, E4 antibodies showed higher prevalence in condyloma lesions (79%; 11/14) than in cervical cancer (60%; 12/20). We also identified 11% (4/38) of healthy individuals as positive for E4 antibodies, which suggests an early immune recognition of this protein. Patients with condyloma and cervical intraepithelial neoplasia (CIN) also showed higher prevalences of Ras antibodies (approximately 40%) than cervical cancer patients (10%; 2/20). By sequencing part of the ras genes and using different Ras antigens, we showed that serum antibodies from patients were not directed to a Ras mutation, since wild type cHa-Ras protein was recognized by these antibodies. In addition, patients positive for Ras antibodies (94%) were also positive for E4 antibodies, suggesting an association between these. The high prevalence of antibodies against Ras and E4 proteins in pre-malignant lesions opens the possibility of using both antibodies as early markers for potential cervical cancer patients. PMID- 10795527 TI - The construction of defective interfering rubella virus particles. AB - Two defective viral vectors containing nucleic acid sequences from rubella virus strains RJ and RS linked to the reporter gene luciferase have been constructed. The vector RNAs are shown to replicate and interfere with wildtype virus in co infected cells. The interference is associated with a polymorphic nucleotide at nt34 in the 5' stem-loop. The use of these constructs as expression vectors is discussed. PMID- 10795528 TI - Complete nucleotide sequence of the genomic RNA of a mild strain of Japanese yam mosaic potyvirus. AB - We determined the complete nucleotide sequence of a mild strain of Japanese yam mosaic potyvirus (JYMV-M) and compared it with the published sequence of severe strain of JYMV (JYMV-J1). The genomic RNA of JYMV-M is 9,760 nucleotides (nts) in length, excluding the poly (A) tail, and encodes a polyprotein of 3,132 amino acids. Among nine potential cleavage sites, only the P1 and NIa recognition sites (between 6K1 and CI) had different sequences from those of JYMV-J1. The data confirm the strain status of these two viruses with 91.1% sequence identity for the polyprotein and approximately 94-97% identities for HC, CI, NIa, NIb and CP. The most divergent products P1 and P3 had 62% and 90% sequence identities respectively. PMID- 10795529 TI - Molecular characterization and coat protein serology of watermelon leaf mottle virus (Potyvirus). AB - A cDNA library was generated from purified RNA of watermelon leaf mottle virus (WLMV) (Genus Potyvirus). Two overlapping clones totaling 2,316 nucleotides at the 3' terminus of the virus were identified by immunoscreening with coat protein antiserum. The sequence analyses of the clones indicated an open reading frame (ORF) of 2,050 nucleotides which encoded part of the replicase and the coat protein, a 243-nucleotide non-coding region (3'UTR), and 23 adenine residues of the poly (A) tail. The taxonomic status of WLMV was determined by comparisons of the sequence of the cloned coat protein gene and 3'UTR with potyvirus sequences obtained from GenBank. The nucleotide sequence identities of WLMV compared with 17 other potyviruses ranged from 55.6 to 63.5% for the coat protein, and from 37.2 to 48.3% for the 3'UTR. Phylogenetic analyses of the coat protein region and the 3'UTR indicated that WLMV did not cluster with other potyviruses in a clade with high bootstrap support. The coat protein gene was expressed in Escherichia coli and a polyclonal antiserum was prepared to the expressed coat protein. In immunodiffusion tests, WLMV was found to be serologically distinct from papaya ringspot virus type W, watermelon mosaic virus 2, zucchini yellow mosaic virus, and Moroccan watermelon mosaic virus. In Western blots and ELISA, serological cross-reactivity with other cucurbit potyviruses was observed. Serological and sequence comparisons indicated that watermelon leaf mottle virus is a distinct member of the Potyvirus genus. PMID- 10795530 TI - The nucleotide sequence and genome organization of Japanese iris necrotic ring virus, a new species in the genus Carmovirus. AB - The genome of Japanese iris necrotic ring virus (JINRV) consists of a positive sense ssRNA of 4014 nucleotides with six major open reading frames (ORFs). A 5' non-coding region of 31 nucleotides precedes the first initiation codon. Like Carnation mottle virus (CarMV), the 5'-proximal three ORFs encode a 26 kDa protein (p26) and two readthrough proteins, i.e. an 85 kDa putative RNA replicase (p85) and a 99 kDa protein (p99). The central ORF encodes a small 8 kDa protein (p8). The 3'-proximal ORF encodes a 38 kDa capsid protein (p38). Another ORF encoding a 12 kDa protein (p12) overlaps the p99 ORF.JINRV RNA treated with bacterial alkaline phosphatase and tobacco acid pyrophosphatase could not be ligated to an oligoribonucleotide using T4 RNA ligase, indicating that the 5' end of the viral RNA is uncapped. The 3' end is not polyadenylated. Comparison of the genomic organization and the predicted amino acid sequences with those of other viruses confirmed that JINRV should be classified as a member of the genus Carmovirus, family Tombusviridae. PMID- 10795531 TI - Middle cerebral artery variations detected by magnetic resonance angiography. AB - Middle cerebral artery (MCA) variations are found incidentally on cranial magnetic resonance angiography (MRA). Our goal was to examine the incidence and types of MCA variations detected by MRA. Between April 1996 and March 1999, cranial MRA was performed in 432 cases at our institution. Most of the patients examined had or were suspected to have cerebrovascular disease. After excluding 7 patients with moyamoya disease, we retrospectively reviewed 425 MRA results. A 1.5-T scanner was used in all studies, and maximum-intensity projection images obtained using the three-dimensional time-of-flight technique were displayed stereoscopically. In the 425 patients MRA revealed 16 anomalous MCAs, including 9 duplicated MCAs, 5 accessory MCAs, and 2 fenestrated MCAs, which is a rate of 3.8%. Thus, although the clinical significance is not great, we found a relatively high incidence of anomalous MCAs on MRA. We stress that knowledge and recognition of these variations are useful and important during the interpretation of cranial MRA. PMID- 10795532 TI - Three-dimensional reconstructed rotational digital subtraction angiography in planning treatment of intracranial aneurysms. AB - We evaluated three-dimensional (3D) reconstructions of 200 degrees rotational digital subtraction angiography (DSA) images for their contributions to improving the safety of endovascular embolization of intracranial aneurysms. Standard DSA and 200 degrees rotational DSA were performed in 40 adult patients (aged 21-77 years) with 45 intracranial aneurysms. Information obtainable from standard DSA and 3D-DSA images about aneurysm shape and size was compared. In 40 (89%) of the 45 aneurysms 3D-DSA gave additional information about the anatomy of the aneurysm. In 17 (43 %) of these cases aneurysm anatomy could be visualized better on 3D-DSA than on standard DSA images. In three cases only 3D-DSA images showed blood vessels originating from the aneurysm. Reconstructed 3D images were also helpful in visualizing partially clipped aneurysms. On maximum-intensity projection images it was even possible to depict previously embolized aneurysms. Blood vessels originating from the aneurysm are visible on 3D-DSA images, and even previously clipped aneurysms can be visualized well. Rotational DSA with 3D reconstruction is a helpful tool in the assessment of intracranial aneurysms. PMID- 10795533 TI - Recent experience with Moyamoya disease in Turkey. AB - A series of moyamoya patients is presented. Angiographic findings, outcome of revascularization surgery and a young case with moyamoya disease and hyperphosphatemia are reported. Thirteen patients (6 males and 7 females; age range 2-50 years) were included in the study group. Findings of the patients at presentation were intracranial haemorrhage in two adult cases and sequelae of cerebral ischemia in the rest of the group. One young girl had hyperphosphataemia. Angiography showed distal internal carotid or proximal anterior and middle cerebral artery stenosis, unique collaterals, microaneurysm of the posterior lateral choroidal artery and flow-related changes in the posterior circulation. In 3 patients, encephalo-duro-arterio-synangiosis (EDAS) and burrholes were performed at surgery. Follow-up angiograms of these patients showed revascularization. Moyamoya, a rare but potentially devastating disease, must be addressed as a cause of haemorrhagic and ischaemic cerebral events. PMID- 10795534 TI - Optic nerve aspergillosis: report of a case diagnosed by fine-needle aspiration biopsy. AB - The aim of this study was to describe the fine-needle aspiration biopsy (FNAB) procedure in patient with a optic nerve lesion. A case of optic nerve and brain aspergillosis, an infrequent condition that can mimic tumor or tumor-like lesions, is reported. The patient was studied by CT and MRI and definite diagnosis was established by means of an ultrasound-guided FNAB. Specimen preparation revealed the presence of multiple hyphae of Aspergillus fumigatus in the optic nerve lesion. The FNAB procedure is a safe and reliable diagnostic method for suspected intraorbitary tumors and tumor-like conditions especially when other less-invasive modalities have failed to establish the diagnosis and when cytologic confirmation is needed to implement aggressive therapy. PMID- 10795535 TI - MRI of the brain in cerebrotendinous xanthomatosis (van Bogaert-Scherer-Epstein disease). AB - The clinical, biochemical and magnetic resonance imaging findings of two patients with cerebrotendinous xanthomatosis are reported. This is a rare hereditary disease. Early recognition of this entity is important in view of the existing treatment possibilities. Magnetic resonance imaging findings typically include a bilateral and almost symmetrical increase of the signal intensity on the T2 weighted images in the cerebellar and periventricular cerebral white matter, the basal ganglia, the dentate nuclei and the brainstem as well as cerebellar and cerebral atrophy. PMID- 10795536 TI - Rapid MRI assessment of renal space-occupying lesions. AB - The aim of this study was to determine whether rapidly acquired MRI sequences, taking less than 5 min imaging time, can accurately characterise renal masses. All patients found to have a renal space-occupying lesion on CT or asked to participate in a prospective study using rapidly acquired MRI. The MRI technique was performed on a GE Signa (General Electric, Milwaukee, Wis.) 1.5 T magnet using breath-hold coronal and axial T1 GRASS (fast spoiled gradient-recalled acquisition into steady state, FSPGR30/90) and axial T2 fast spin-echo sequences. The results were analysed by two radiologists unaware of the CT or ultrasound findings. The CT/US was independently viewed by a third radiologist. Lesions were characterised as simple cysts, indeterminate or solid. The MR and CT/US results were correlated and the sensitivity and specificity of MR for the characterisation of simple cysts and solid renal masses calculated. A total of 144 lesions (68 patients; 29 women 39 men, age range 32-78 years, average age 60 years) were studied: 111 simple cysts; 3 hyperdense cysts; 26 renal cell carcinomas; and 4 indeterminate lesions on CT/US. There was agreement between the CT/US and MRI in 82% of cases. All renal cell carcinomas were correctly characterised on MRI. Of simple cysts, 79% were correctly identified using this technique. Breath-hold MRI performed in less than 5 min can accurately characterise the majority of renal masses. It is 100% sensitive in the characterisation of renal carcinoma, and it correctly identified approximately 80% of simple cysts. If used at the time a renal mass is detected on MRI, it would reduce the need for further investigation of the majority of incidentally detected lesions which are simple cysts. PMID- 10795537 TI - Right adrenal hematoma following hepatectomy. AB - We present the findings and possible causes in three cases of postoperative adrenal hematomas. In 16 cases of 45 consecutive patients, following segmental right or left lobe hepatectomy, CT was performed in order to evaluate possible fluid collection or other complications. In all cases imaging findings and a correlation with preoperative CT scans were done. Follow up CT examinations were also reviewed. In three cases solid suprarenal masses with attenuation values consistent with adrenal hematomas were found. Preoperative scans at the same level indicated normal adrenal glands. Follow-up scans revealed the hematoma, stable in size, for up to 12 weeks, although lower attenuation values were evident. Right adrenal hematoma is a possible postoperative complication following hepatectomy; if it remains stable in size, it can be left alone. PMID- 10795538 TI - Xanthogranulomatous pyelonephritis: MRI findings in the diffuse and the focal type. AB - Two cases of xanthogranulomatous pyelonephritis (XGP) are presented with emphasis on MR appearances. One case is the diffuse type of XGP secondary to chronic obstruction caused by transitional cell carcinoma of the renal pelvis. The other case is the focal or "tumefactive" type of XGP which mimics renal cell carcinoma. PMID- 10795539 TI - Primary seminoma of the retroperitoneum. AB - A rare case of primary seminoma in the retroperitoneum is described. Ultrasound detected a huge hypoechoic mass and correlative computed tomography showed a midline, homogeneous, lobulated mass in a man who presented for anuria. PMID- 10795540 TI - Preoperative embolization of bone metastases from renal cell carcinoma. AB - The purpose of this study was to correlate the effectiveness of preoperative embolization with the blood loss and transfusion requirement during surgery for bone metastases from renal cell carcinoma. Twenty-eight preoperative embolizations in 26 patients with renal cell carcinoma metastatic to bone were retrospectively evaluated and divided into two groups: Group A included the embolizations that resulted in complete devascularization of the lesion as defined by the post-embolization arteriograms, and group B included those with an incomplete result. The two groups were compared with regard to blood loss and transfusion requirement during surgery, by unpaired two-tailed Student's t-test. Where complete embolization was effected (group A, 10 cases), there was a mean blood loss of 535 +/- 390 ml. When a less than complete embolization was achieved (group B, 18 cases), the mean blood loss was 1.247 +/- 1.047 ml (p = 0.049). The red blood cell transfusion in group A was 1.3 +/- 1 units, whereas in group B it was 2.4 +/- 1.2 (p = 0.03). Preoperative embolization of bone metastases from renal cell carcinoma with subsequent complete devascularization leads to significant reduction of blood loss during surgery. Interventional radiologists should pursue and embolize every feeder to the metastasis, because any less than complete devascularization increases the amount of blood loss and the amount of red blood cell transfusion during surgery. PMID- 10795541 TI - Functional imaging of the thoracic outlet syndrome in an open MR scanner. AB - Symptoms due to thoracic outlet syndrome may present only in abduction, a position that cannot be investigated in conventional MR scanners. Therefore, this study was initiated to test MRI in an open magnet as a method for diagnosis of thoracic outlet syndrome. Ten volunteers and 7 patients with a clinical suspicion of thoracic outlet syndrome were investigated at 0.5 T in an open MR scanner. Sagittal 3D SPGR acquisitions were made in 0 and 90 degrees abduction. In the patients, a similar data set was also obtained in maximal abduction. To assess compression, the minimum distance between the first rib and the clavicle, measured in a sagittal plane, was determined. In the neutral position, no significant difference was found between patients and controls. In 90 degrees abduction, the patients had significantly smaller distance between rib and clavicle than the controls (14 vs 29 mm; p < 0.01). On coronal reformatted images, the compression of the brachial plexus could often be visualised in abduction. Functional MR examination seems to be a useful diagnostic tool in thoracic outlet syndrome. Examination in abduction, which is feasible in an open scanner, is essential for the diagnosis. PMID- 10795542 TI - Evaluation of radiolucency condition in total hip arthroplasty: a statistical comparison of the diagnostic capability of digitised image vs. conventional X-ray film. AB - This work analyses the diagnostic capability of radiographic images taken from patients with total hip arthroplasty and visualised on monitor. Images were obtained with digital acquisition of conventional X-ray films. The investigated pathology is the absence of direct contact between bone and prosthesis (radiolucency). Three senior orthopaedists defined the diagnostic "truth" on well defined regions of interest on 22 conventional X-ray films of total hip arthroplasty, obtaining a total of 110 reference ratings. Films were digitised by use of an X-ray scanner. Four readers evaluated the X-ray images, applying conventional and monitor visualisation. To show any difference between ratings on film and ratings on monitor a sensitivity, specificity and accuracy study jointly with a receiver operating characteristics (ROC) study were performed for each reader and for all combined readings. The intra-observer reproducibility of the radiographic protocol was equal to 87% and the inter-observer one was in the range 85-92%. The sensitivity, specificity and accuracy study together with the ROC analysis did not show significant differences between the two evaluation modes. The evaluation of radiolucency from digitised X-ray films visualised on a monitor resulted statistically comparable with the conventional evaluation on X ray films. PMID- 10795543 TI - Age-related changes assessed by peripheral QCT in healthy Italian women. AB - The purpose of this study was to describe the normal cross-sectional pattern of radial bone loss associated with aging in healthy women and to generate a normative database using peripheral quantitative computed tomography (pQCT). Subjects with suspected conditions affecting bone metabolism or receiving any drugs affecting bone mineralization were excluded. The trabecular bone mineral density (BMD) and the total bone density of the ultradistal radius at the nondominant forearm was measured using the Norland-Stratec XCT-960 pQCT scanner in 386 healthy pre-, peri-, and postmenopausal females aged 15-81 years. The long term in vivo precision error was 1.6 % CV (coefficient of variation) for trabecular and 0.8% CV for total BMD measurements. The highest value of trabecular and total BMD measured was observed at the age group 15-39 years. Beyond these ages both trabecular and total BMD showed a linear decline with aging, decreasing by an overall slope of -1.28 and -0.55 mg/cm3 per year for total and trabecular BMD measurements, respectively. The test of parallelism between the regression slopes of the peri- and postmenopausal women showed a statistically significant difference for total BMD measurement (p = 0.003). Measurement of total and trabecular BMD was not influenced by weight, height or body mass index, but it was correlated with natural logarithm of years since menopause. We conclude that pQCT of the ultradistal radius is a precise method for measuring the true volumetric BMD and for detecting age-related bone loss in the trabecular and total bone of female subjects encompassing the adult age range and menopausal status. PMID- 10795544 TI - MR imaging of lumbar facet joint synovial cysts. AB - The increasing application of magnetic resonance (MR) imaging of the spine has raised the awareness of lumbar facet synovial cysts (LFSC). This well recognised, yet uncommon condition, presents with low back pain and radiculopathy due to the presence of an extradural mass. The commonest affected level is L4/5 with a mild degenerative spondylolisthesis a frequent associated finding. MR imaging is the technique of choice to detect and diagnose a LFSC. This pictorial essay, drawing on experience of 43 cases seen in 40 patients, illustrates the spectrum of appearances that can be encountered and suggest differing causes for the variable signal characteristics exhibited. Computed tomography (CT) can be of value in some cases to aid interpretation of the MR images. In addition, CT facet arthrography by injection of air or iodinated non-ionic contrast medium may be used to confirm the diagnosis in doubtful cases as well as noting whether the patients presenting symptoms can be provoked. A comprehensive review of the existing literature is presented. PMID- 10795545 TI - Acetabular ossicles: normal variant or disease entity? AB - Ossicles located in the acetabular fossa may confuse diagnostic and therapeutic work-up. An accessory ossification centre may persist unfused as an os acetabuli centrale which is surrounded by intact hyaline cartilage representing an anatomic variant. Bone islands located in the pillars of the acetabulum can project into the acetabular fossa simulating acetabular ossicles. Osteochondrosis dissecans, posttraumatic articular bodies, degenerative disease and other rare lesions may be responsible for clinical symptoms and are of similar appearance than anatomic variants. Plain film radiography, X-ray tomography, CT and MRI are used to categorize these lesions. MRI is very valuable to assess cartilage integrity in a noninvasive way, but arthro-CT or arthro-MRI have to be used in unclear cases. Therefore the purpose of this presentation is to discuss the appearance, the possible etiology and the differential diagnosis of acetabular ossicles and how they can be evaluated to avoid an unnecessary arthrotomy. PMID- 10795546 TI - Visualization and quantification of coronary calcifications with electron beam and spiral computed tomography. AB - This contribution reviews the pathology and morphology of coronary calcifications. It summarizes the indications for investigation of the coronary arteries. The standard protocols for scan acquisition using electron beam and conventional computed tomography are described as well as various methods for evaluation such as the traditional Agatston scoring method and the newer three dimensional scoring algorithms. Guidelines for interpreting scores are also reviewed. Major limitations of the reproducibility of the calcium score measurement are summarized. Future aspects of multirow-detector spiral computed tomography with retrospective electrocardiographic triggering for quantifying coronary calcium are discussed. PMID- 10795547 TI - Endoluminal grafting of abdominal aortic aneurysms: experience with the Talent endoluminal stent graft. AB - The aim of this study was to evaluate the Talent endoluminal stent graft (TESG) in treating abdominal aortic aneurysms (AAA). The TESG is a polyester-covered nitinol endograft (proximal diameters 20-38 mm and iliac limb diameters 8-22 mm). Twenty-two men were treated with the TESG via bilateral femoral arteriotomies. Pre-implantation, coil embolization of various vessels arising from the aneurysm was performed in 6 patients. Plain radiographs and spiral CT angiograms (CTA) were carried out at 7 days, 3, 6, and 12 months following TESG implantation or re intervention. Median aortic and iliac diameters were 27 mm (range 20-34 mm) and 14 mm (range 10-19 mm). The corresponding graft diameters were 30 mm (range 24-38 mm) and 14 mm (range 12-20 mm). No patient was rejected purely on the basis of too large aortic or iliac diameters. Eight patients required custom-made grafts. Graft implantation was successful in all patients. There were no blood transfusions, distal embolic episodes, or conversions to open surgery. Re intervention was necessary in 1 patient. Complications included one fatal myocardial infarction, one inguinal hematoma, and two superficial wound infections. The aneurysm thrombosed completely following implantation in 14 patients and at 3 or 6 months in 4 other patients. One patient with endoleak is awaiting his 3-month control and 2 patients show tiny endoleaks but reduction of aneurysm size. The mean aneurysm size decreased significantly from 58 +/- 10 to 53 +/- 13 mm (p < 0.0005). Due to the large sizes available and the option of custom-made grafts, the TESG helps widen the spectrum of patients who can be treated with endoluminal grafting. The treatment is associated with a significant reduction in aneurysm size. PMID- 10795548 TI - Intra-arterial Doppler flowmetry in the superficial femoral artery following angioplasty. AB - The aim of the study was to assess the diagnostic value of an intravascular Doppler guidewire in patients with peripheral percutaneous angioplasty (PTA). The prognostic value was also evaluated. Measurements were done prior and following angioplasty in 22 patients with peripheral arterial occlusive disease. As additional therapy, stent insertion and peripheral (Aa. poplitea Tll/tibial) angioplasty was performed (4 patients per group). For stress testing, adenosinetriphosphate (ATP) was given intra-arterially. Follow-up was performed by angiography, colour-coded duplex ultrasound or judged by unequivocal clinical stage at follow-up to 13 months. Average (APV) and maximal peak velocity (MPV) increased following PTA, after additional treatment (peripheral PTA or stent), and after intra-arterial application of a vasodilator. Patients with peripheral lesions had markedly lower velocities prior treatment and following PTA after vasodilatation. Following peripheral PTA, the values were similar to the patients with PTA alone. Velocities after stenting were markedly increased in the stress condition. Of the 22 patients, 7 had a recurrent disease. The latter patients had higher velocities at rest prior to and following PTA. In stented lesions higher velocities seem to be linked with a worse outcome. The ratio between velocity prior to and after the application of the vasodilator seems to be of diagnostic importance. A ratio of 1.9 or more was of positive prognostic value. The Doppler guidewire is a practical and valuable tool in assessing technical success after angioplasty of peripheral lesions, critical or morphologically worse lesions. In our study the decision for stent application was made on the morphological image; however, increased velocity and changes in phasicity substantiated our decisions. Increased ratios prior to and after vasodilation (flow reserve) are of prognostic value and therefore suitable as indication for stent placement or tibial angioplasty. PMID- 10795549 TI - MR diagnosis of a congenital abnormality of the thoracic aorta with an aneurysm of the right subclavian artery presenting as a Horner's syndrome in an adult. AB - Congenital abnormality of the aortic arch is a diagnosis made most of the time incidentally in childhood, unless dysphagia or respiratory disorders occur before. A case of a complex aortic arch anomaly with an aneurysm of the right subclavian artery presenting as an isolated Horner's syndrome in an adult is reported herein. Magnetic resonance imaging led to this very unusual diagnosis. PMID- 10795550 TI - Rectal tumour staging: MR imaging using pelvic phased-array and endorectal coils vs endoscopic ultrasonography. AB - The aim of this study was to compare MR imaging and endoscopic ultrasonography (EUS) for the local staging of rectal tumours. Forty-nine patients were examined on a 1.5-T MR unit using either a pelvic phased-array coil (n = 37) alone or combined with an endorectal coil (n = 12). Sagittal and axial sequences with T2 weighted fast spin-echo and axial T1-weighted spin-echo techniques were employed. The EUS technique was performed using a flexible endosonoscope. The results were compared with findings at histopathological sectioning of the specimen. The T stage on MR correlated with histopathology in 32 of 49 patients and on EUS in 29 of 49 patients. The N-stage on MR correlated with histopathology in 22 of 49 patients and on EUS in 26 of 49 patients. Tumour penetration of the rectal wall was predicted by MR with 86 % sensitivity and 65 % specificity, and by EUS with 89% sensitivity and 33% specificity. Preoperative radiotherapy was administered to 40 of the patients after the examinations which may explain some of the overstaging by MR and EUS. Three patients with surgically and histopathologically confirmed invasion of neighbouring organs in the pelvis were detected preoperatively on MR but none on EUS. Tumour penetration of the rectal wall and local lymph node metastases cannot accurately be predicted with MR or EUS. Magnetic resonance, however, seems to be more useful for preoperative identification of clinically occult advanced disease. PMID- 10795551 TI - Radiological anatomy of the groin region. AB - The purpose of this paper is to give an overview of the anatomy of the inguinal region, and to discuss the value of various imaging modalities in the diagnosis of groin hernias. After description of the gross anatomy of the groin, attention is focused on the anatomic features of conventional herniography, US, CT, and MRI. Advantages, disadvantages, and accuracy of each technique is discussed briefly. PMID- 10795552 TI - Helical CT appearance of glomus tumor of the stomach. AB - Glomus tumor occurs only rarely in the stomach. This benign intramural mass is located most frequently in the distal half of the stomach. We experienced two cases of glomus tumor of the stomach which were examined with helical CT and were confirmed pathologically. Both tumors were well-marginated solitary lesions, located in the gastric antrum. On the early-phase helical CT, an intact overlying mucosa was demonstrated in both cases, and both tumors showed a dense homogeneous contrast enhancement, which persisted to the delayed phase. PMID- 10795553 TI - Appendicitis after appendectomy: CT diagnosis. AB - This article presents a case of appendicitis 7 years after open appendectomy. Together with the apparent CT findings we discuss the current literature of this issue. PMID- 10795554 TI - Frequency of pancreatitis after endoscopic retrograde cholangiopancreatography with iopromid or iotrolan: a randomized trial. AB - Blood isotone contrast media is considered to be less toxic to vascular and pancreatic duct endothelium than high-osmolar contrast media. In this study we assessed the impact of a low-osmolar contrast agent compared with a blood isotone product on pancreatic damage induced by endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic retrograde sphincterotomy (EST). In a prospective trial 42 consecutive ERCP/EST patients were randomized to receive either iopromid, a low-osmolar non-ionic contrast agent (770 mosmol/kg H2O), or iotrolan, a blood-isotone non-ionic product (320 mosmol/kg H2O). The endoscopies were performed by two experienced endoscopists. Forty patients were included in the study. Blood samples were collected before and 40 min, 2, 4, 6 and 24 h after the endoscopic procedure. Samples were analysed for pancreatic serum enzymes, acute-phase proteins and blood counts. A clinical pain score was investigated. Post-ERCP pancreatitis was diagnosed in 2 patients in the iopromid group and in 5 patients in the iotrolan group. There was no significant difference between groups in the time course of pancreatic serum enzymes, acute phase proteins or in the pain score. Due to the small number of patients in this study, only stronger differences caused by the two contrast media could have led to statistically significant results. We did not observe statistically significant differences in comparing iotrolan and iopromid concerning ERCP/EST induced pancreatic damage. PMID- 10795555 TI - Evaluation of CT findings for diagnosis of pleural effusions. AB - Computed tomography studies are usually used to assess patients with pleural effusions, and radiologists should be aware of the significance of different CT findings for the diagnosis of the effusion. The purpose of this study was to evaluate CT findings for etiological diagnosis of pleural effusions. Contrast enhanced CT of the chest of 211 patients with pleural effusion of definite diagnosis were evaluated. The CT images were evaluated for the presence and extent of pleural effusion, thickening or nodules, extrapleural fat and other changes in the mediastinum or lung. The CT scans were read by two independent observers and correlation between them was evaluated. Comparison of CT findings between benign and malignant effusions, between exudates and transudates, and between empyemas and the other parapneumonic effusions were carried out. Kappa values for most CT findings were > 0.85. Loculation, pleural thickening, pleural nodules, and extrapleural fat of increased density were only present in exudative effusions. Multiple pleural nodules and nodular pleural thickening were the only pleural findings limited to malignant pleural effusions. The signs were also more frequently seen in empyemas than in other parapneumonic effusions. Computed tomography findings can help to distinguish between transudates and exudates. Although there is some overlap between benign and malignant pleural effusions, pleural nodules and nodular pleural thickening were present almost exclusively in the latter. Although differences between CT findings of empyemas and the other parapneumonic effusions exist, there is no finding which can definitely differentiate between them. PMID- 10795556 TI - Mediastinal myelolipoma: CT and MRI appearances. AB - A 72-year-old man presented with a mediastinal mass on chest radiograph. Computed tomography and MR imaging showed that the mass consisted of both fatty and small nodular soft tissue components, highly suggestive of an extramedullary hematopoiesis or a myelolipoma. A CT-guided needle biopsy was next performed and confirmed the diagnosis. We discuss the CT and MR imaging appearances es of this tumor and usefulness of a CT-guided needle biopsy to avoid surgery in asymptomatic patients. PMID- 10795557 TI - Prostate specific antigen: an opinion on its value to the radiologist. PMID- 10795558 TI - Child neurology. PMID- 10795559 TI - Development of the corticospinal system and hand motor function: central conduction times and motor performance tests. AB - Maturation of the corticospinal (CS) tract and hand motor function provide paradigms for central nervous system development. In this study, involving 112 participants (aged from 0.2 to 30 years), we evaluated central motor conduction times (CMCT) obtained with transcranial magnetic stimulation (TMS) during preinnervation conditions of facilitation and relaxation. Auditory reaction time, velocity of a ballistic movement of the arm, finger tapping, diadochokinesis, and fine motor visuomanual tracking were also examined. The maturation profiles for every parameter were calculated. CMCTs for the different preinnervation conditions reached adult values at different times and this could be explained by maturation of excitability at the cortical and spinal level. A stable phase for CMCTs and reaction time was reached during childhood. Parameters which measured motor speed and skill indicated that the development of these continued into adulthood. The maturation of the fast CS tract seems to be completed before the acquisition of the related motor performance has been accomplished. In conclusion, we could demonstrate that data from several neurophysiological methods can be combined and used to study the maturation of the function of the nervous system. This approach could allow appraisal of pathological conditions that show parallels with omissions or lack of developmental progress. PMID- 10795560 TI - Impaired force coordination during object release in children with hemiplegic cerebral palsy. AB - Recent studies on hand motor control in children with cerebral palsy (CP) have focused on the coordination of fingertip forces during the grasping and lifting of objects. However, little is known about the ability to replace and release an object from grasp, a function that is just as important for fine dexterity. The present study examines the coordination of fingertip forces during the replacement and release of an object (either 200 g or 400 g) from grasp in 14 children (aged between 7 and 13 years) with hemiplegic CP and in 14 age-matched typically developing children. The results indicate that children with hemiplegic CP abruptly replaced the object but had a prolonged and uncoordinated release of the grasp. Unlike what was seen in the control children, the grip and load forces decreased sequentially in the children with CP. However, all the children could appropriately scale the rate of force decrease based on somatosensory weight related information from the ongoing lift. The results provide further information about the impaired hand skills in children with hemiplegic CP. PMID- 10795561 TI - Identification of neonates at risk of developing feeding problems in infancy. AB - The increased survival of sick and preterm neonates may be associated with long term problems which must be recognised and managed if outcome is to be optimised. In a prospective study of 35 neonates (median gestational age at birth 34 weeks) admitted to a neonatal intensive care unit over a 3-month period, we have documented a high incidence (14 of 35) of immature or abnormal feeding patterns when infants were assessed at 36 to 40 weeks postmenstrual age. Neonates with prolonged respiratory support and delayed enteral and oral feeding were most affected. Compared with neonates who have normal initial feeding assessments, neonates with disorganised or dysfunctional feeding were six times more likely to vomit and three times more likely to cough when offered solid food at 6 months of age. At 12 months of age significant differences were also found in tolerating lumpy food and enjoying mealtimes. We hypothesise that these feeding problems contribute to failure to thrive and psychosocial distress after discharge from the neonatal unit and propose potential neonatal measures to reduce their incidence. PMID- 10795562 TI - Visual evoked potentials in young persons with newly diagnosed diabetes: a long term follow-up. AB - To evaluate the presence of electrophysiological abnormalities in the visual function of young persons with diabetes, visual evoked potentials were recorded, in basal conditions and after photostress, in 30 patients with newly diagnosed insulin-dependent diabetes mellitus. Their mean age was 17.6 years (3.6 SD), and their glycosated haemoglobin (HbAlc) was 9.4% (1.6 SD). Thirty healthy age- and sex-matched individuals were evaluated as the control group. This study showed that the P100 latency was significantly delayed in patients with diabetes compared with the control group (p<0.01), while the N75 to P100 amplitude was similar in both groups. These measurements were repeated after 6 months, when all participants with diabetes had achieved good metabolic control (HbAlc 7.2% [1.5 SD]). At this second evaluation a complete normalisation of all parameters was observed. These findings suggest that early functional abnormalities of the optic nerve can be detected at onset of diabetes, and that glycaemic control reverses these abnormalities. PMID- 10795563 TI - Comparison of the Gross Motor Function Measure and Paediatric Evaluation of Disability Inventory in assessing motor function in children undergoing selective dorsal rhizotomy. AB - This study was designed to compare assessment with the functional outcome measures Gross Motor Function Measure (GMFM) and Pediatric Evaluation of Disability Inventory (PEDI) over time, in children with cerebral palsy (CP) undergoing selective dorsal rhizotomy combined with individualised physiotherapeutic interventions. Using the Gross Motor Function Classification System (GMFCS), 18 children with spastic diplegia were divided into two groups according to age-related severity of motor function impairment. Data were collected preoperatively, and at 6 and 12 months postoperatively. Both instruments were sensitive to changes in function over time in the series as a whole and in the group with milder impairment, although the PEDI detected significant changes earlier. In the group with more severe impairment, changes in function were detected only with the PEDI, not with the GMFM. Thus, the instruments are to be considered complementary tests, because they measure different aspects of function. PMID- 10795564 TI - Development of ideomotor praxis representation. AB - To create a short screening instrument to investigate the development of ideomotor praxis representation (IPR), 357 normally developing children between the ages of 2 1/2 and 9 1/2 years were investigated. The IPR screening consisted of six mimed actions which had to be performed on verbal request. The greatest change in the representational style is the transition from performing the action using the body as the object to performing with an imaginary object (i.e. symbolic representation). This transition suggests a development from a concrete and egocentric level of performance to an abstract and allocentric level. Before the age of 6 years differences are evident between self-directed and externally directed gestures. The level of symbolic representation has to be dissociated from the degree of motor precision, which shows hardly any improvement after 6 years of age. With the proposed model one may interpret the level of IPR in clinical practice. IPR screening is clinically relevant from the age of 4 1/2 years onwards. PMID- 10795565 TI - Dysfunction of the auditory cortex persists in infants with certain cleft types. AB - Language and learning disabilities occur in almost half of individuals with oral clefts. The characteristics of these cognitive dysfunctions vary according to the cleft type, and the mechanisms underlying the relation between cleft type, cognitive dysfunction, and cleft-caused middle-ear disease are unknown. This study investigates preattentive auditory discrimination, which plays a significant role in language acquisition and usage, in infants with different cleft types. A mismatch negativity (MMN) component of brain evoked potentials, which indexes preconscious sound discrimination, and brain responses to rare sine wave tones were recorded in 12 healthy infants and 32 infants with oral clefts at the ages of 0 and 6 months. Infants with clefts were subdivided into two categories: those with cleft lip and palate (CLP) (n=11 at birth, n=6 at the age of 6 months) and those with cleft palate only (CPO) (n=17 at birth, n=8 at the age of 6 months). At both ages, brain responses to rare sounds tended to be smaller in both cleft subgroups than in healthy peers. However, in the latency range of 300 to 500 ms, the MMN was significantly smaller in infants with CPO. In infants with CLP, the MMN was comparable to that of healthy infants. Differences in auditory discrimination between infants with CLP and CPO, as reflected by MMN, were detectable at birth and persisted into later infancy. This pattern parallels known behavioural differences between children with these cleft types. Brain responses to rare sounds, in contrast, had no differentiative power with respect to the cleft type. PMID- 10795566 TI - Prevalence of psychiatric diagnoses in children with mental retardation: data from a population-based study. AB - The main purpose of the study was to estimate the prevalence of psychiatric diagnoses in children with mental retardation (MR) (IQ < or = 70). All children born between 1980 and 1985 (N=30037) in Akershus County, Norway, were screened for possible MR and assessed with either IQ tests or standardized developmental tests. A total of 178 children, 79 with severe mental retardation (SMR) (IQ<50) and 99 with mild mental retardation (MMR) (IQ 50 to 70) were included for further study. Psychiatric symptomatology was assessed as a standard part of the neurodevelopmental examination, which included a semistructured parent interview, a clinical child interview, and retrieval of the charts of previous child psychiatric examinations. Psychiatric diagnoses were classified according to the International Classification of Disease (ICD-10). In total, 65 (37%) of the total population with MR (95% confidence intervals 29 to 44) were registered to have psychiatric diagnoses, the most common being hyperkinesia (n=28) and pervasive developmental disorder (n=15). Psychiatric diagnoses were present in 42% of the population with SMR and 33% of the population with MMR (p=0.4). Of all children found to have a psychiatric diagnosis, approximately one-third had previously been examined by a child psychiatrist and indicated a previously unrecognized need for these services to children with MR. PMID- 10795567 TI - Air swallowing in Rett syndrome. AB - The possible causes of excessive swallowing of air leading to bloating, which is common in Rett syndrome (RS), were investigated during feeding and at rest. Seven individuals with RS aged between 4 and 33 years (three with air bloat) underwent feeding videoflouroscopy and concurrent respiration monitoring. The results were compared with a randomly selected group of 11 individuals, aged between 2 and 16 years, with quadriplegic cerebral palsy and feeding problems, some of whom had mild air bloat. All individuals from both groups had isolated pharyngeal swallows and several mouth breathed; this may account for some air swallowing but not the severe air bloat characteristic of RS. Thirty-three individuals with RS aged between 3 and 44 years were monitored for nasal respiration, chest movements, swallowing, and vocal cord position at rest (between feeding). Twenty had air bloat, 17 of whom swallowed air during breath-holding in the same way, and three gulped air during hyperventilation. Of the 13 without air bloat, eight did not have recurrent breath-holding and five did, but without concurrent air swallowing. Several methods for reducing air swallowing in apnoea were investigated. The most successful was a dummy with an air leak, but this was poorly tolerated and could only be used for short periods of time. Apnoeas and air bloat are often worse when individuals are distressed and may in some individuals be reduced by anxiolytic medications. PMID- 10795568 TI - Behavioural treatment of urinary incontinence and encopresis in children with learning disabilities: transfer of stimulus control. AB - Urinary and faecal incontinence present a considerable problem in people with learning disabilities, despite the general effectiveness of behavioural techniques in continence training. Children with learning disabilities and obsessional behaviour may be particularly resistant to toilet training, even where relatively cognitively able, and often despite a substantial degree of control over their eliminatory functions. Their resistance may be more appropriately regarded as a challenging behaviour and their incontinence better explained by factors other than a simple failure to learn. A 'stimulus-control' hypothesis proposes that the child's nappy (diaper)/potty/underwear has developed strong stimulus control over the elimination response. This report describes three case studies in which treatment-resistant children, aged between 8 and 12 years, with mild or moderate learning disabilities, were successfully treated for nappy-dependent nocturnal encopresis or diurnal urinary incontinence. The children were routine case referrals for whom previous attempts to train bowel or bladder control had failed. Behavioural techniques, such as 'shaping' (gradually increasing the proximity to the toilet), 'fading' (reducing the presence of the nappy), and rewards for eliminating, effected successful transfer of stimulus control over elimination from nappy to toilet. Treatment times varied, depending on the degree of the child's obsession and resistance to change. PMID- 10795569 TI - Colpocephaly in two siblings: further evidence of a genetic transmission. AB - Colpocephaly is an abnormal congenital enlargement of the occipital horns of the lateral ventricles, which has typically been associated with learning disability, seizures, and motor and visual abnormalities. This study describes familial colpocephaly in two siblings with different fathers in a Dominican family. There were no other neurological problems in their relatives. Child 1 is a 7-year-old girl, who has had partial epileptic seizures since 4 years of age. She attends a mainstream school. She has coloboma and chorioretinic atrophy in her right eye and a right microphthalmia. MRI revealed colpocephaly and hypogenesis of the corpus callosum. Her brother, aged 2 years 8 months showed a mild delay in speech acquisition. He has had three febrile convulsions and two non-febrile tonic clonic seizures since 6 months of age. Neurological and ophthalmological examinations were normal. MRI documented colpocephaly with right occipital horn prevalence. There are only two literature reports of genetically transmitted colpocephaly hypothesised in two identical twins and in two brothers. In the children in this study, a genetic basis for colpocephaly is confirmed. We hypothesised a maternal transmission with X-linked or autosomal dominance with an incomplete penetrance model of inheritance. PMID- 10795570 TI - Cerebellar ataxia and gluten sensitivity: a rare but possible cause of ataxia, even in childhood. PMID- 10795571 TI - Causes of excess mortality in cerebral palsy. PMID- 10795572 TI - The importance of glassy biopolymer components in food. AB - The glassy state is not just important in low moisture and frozen foods, where it influences the physical and chemical stability and the crispness of some foodstuffs: glassy biopolymer components affect the physical properties of most food systems and low moisture biological systems. Glassy foods are not always fragile and crispy in texture. Therefore, the relationship between mechanical behaviour and molecular dynamics in low-moisture biopolymer systems will be considered. Vitrification of a macromolecular system only requires a mutual fixation of a certain proportion of the chain segments. The higher the rigidity of the chains, the lower the number of the chain segments which must be mutually fixed to vitrify the system. Mechanical stress can help the thermal movement to re-activate the motion of mutually fixed segments and involves a long deformation of glassy material. The stress required to effect long deformation presumably increases up to the strength of the material as the temperature decreases from the glass transition to the temperatures of brittleness (crispness). Vitrification of a loaded viscoelastic system results in an accumulation of mechanical energy (memory effect), which can be released (elastic recovery) above the glass transition temperature due to heating and or addition of a plasticizer. The effects of memory and elastic recovery could be of particular importance for producing foodstuffs which change their form, e.g. self-stirring dry foods and drinks on re-hydration in hot water. The importance of glassy biopolymer ingredients from the viewpoint of food formulation and processing is discussed. PMID- 10795573 TI - Tarhana as a traditional Turkish fermented cereal food. Its recipe, production and composition. AB - As a fermented product tarhana is the dry form of yogurt-cereal mixture and represents an important part of the diets of many people in Turkey. It is prepared by mixing wheat flour, yogurt, yeast and a variety of cooked vegetables (tomatoes, onions, green pepper etc.), salt, and spices (mint, paprika) followed by fermentation for one to seven days. Generally one part yogurt is mixed with two parts of wheat flour (w/w). In commercial production there are two methods for tarhana making. First method is called straight method and ingredients in the recipe is mixed and kneaded, fermented, dried and finally sieved. Second method is called sour dough method that contains three steps, each one has a different recipe. Throughout fermentation lactic acid bacteria and yeast give the characteristic taste and flavour of tarhana by producing lactic acid, ethanol, carbondioxide and some other organic compounds. Organic acids composed in fermentation period lower the pH (3.4-4.2), and low moisture content (6-10%) is a poor medium for pathogens and spoilage organisms. The nutrient content of tarhana depends upon yogurt and flour ratios as well as some other ingredients, and it is also considered to be a useful high-protein dietary supplement with average 15% protein content. Addition of set yogurt due to high dry matter content and baker's yeast increase protein content and enhances it's amino acid composition. PMID- 10795574 TI - Emulsification of chemical and enzymatic hydrolysates of beta-lactoglobulin: characterization of the peptides adsorbed at the interface. AB - Bovine beta-Lactoglobulin (BLG) was cleaved by BNPS-skatole (2-(2' nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine), trypsin, or pepsin in 40% ethanol before emulsification with hexadecane in order to characterize the peptides active at the interfaces. The total digests and the different phases obtained after emulsification were analyzed by RP-HPLC to separate the peptides according to their gradual order on a hydrophilicity-to-hydrophobicity scale. In each case, hydrophobic peptides were recovered in the creamed phase and characterized by mass spectrometry and sequencing. After tryptic hydrolysis, short peptides were identified at the interfacial layer as fragments S21-L32, V41 L57, V41-K60, and W61-K70 linked to L149-I162 by a C66-C160 bond. It indicates that the hydrophilic/hydrophobic distribution of the amino acids in the sequence of the fragments is more relevant to adsorption than the length of the peptide. BNPS-skatole and peptic hydrolysis produced larger hydrophobic peptides which were also recovered in the creamed phase of the emulsion and characterized. PMID- 10795575 TI - Effects of selected process parameters in extrusion of yam flour (Dioscorea rotundata) on physicochemical properties of the extrudates. AB - Raw yam (Dioscorea rotundata) flour was cooked and extruded in a Brabender single screw laboratory scale extruder. Response surface methodology using an incomplete factorial design was applied with various combinations of barrel temperature [100, 125, 150 degrees C], feed moisture content [18, 22, 26%] and screw speed [100, 150, 200 rpm]. Initial viscosity at 30 degrees C, water solubility index, expansion and hardness were determined. The highest values of initial viscosity were at the highest barrel temperatures and the highest moisture contents. At high feed moisture content and high barrel temperatures the yam extrudate flour showed the greatest values of water solubility index. The physical properties of the extruded product showed that at high temperature the lower the moisture content the greater the expansion index. Hardness was influenced directly by moisture content and inversely by extrusion temperature. The extrusion of yam flour led to the production of snacks and pre-gelatinized flours of diverse properties. Also extruded yam flour can be successfully used in the preparation of 'futu' (pre-cooked compact dough), a yam-based food, popular in Western Africa. PMID- 10795576 TI - Isolation and characterization of proline iminopeptidase from Propionibacterium freudenreichii ATCC 9614. AB - A dimeric, 90 kDa subunit intracellular proline iminopeptidase from Propionibacterium freudenreichii ATCC 9614 was purified to homogeneity by chromatography on hydroxyapatite, Sephacryl 200, Phenyl Superose and Mono Q. The enzyme was specific on Pro-p-nitroanilide and Pro-X dipeptides. It hydrolyzed 2 fragments of hormone oligopeptides with an N-terminal proline: bradykinin, f2-7 and substance P, f4-11. A number of oligopeptides containing 5-11 amino acids residues and proline at the penultimate position from N-terminus or other internal position were not hydrolyzed. The enzyme was most active at pH 7-7.5 and at 37-40 degrees C but it retained 9% of maximal activity at pH 5.5 and >12% of maximal activity at 10 or 60 degrees C. The enzyme was inhibited strongly by the serine protease inhibitor 3,4-dichloroisocoumarin, and stimulated markedly by 1 mol/l of NaCl. The results indicate that the enzyme may lead to the accumulation of proline from dipeptides and oligopeptides during the ripening of cheese. PMID- 10795577 TI - Moisture sorption isotherms of cereals at different temperatures. AB - In this research, moisture sorption isotherms of wheat (Kink and Lancer) barley, rye, oat and corn were determined at 20, 25, 35, 50 and 70 degrees C. The sorption isotherm curves of all cereal samples showed the characteristics of type II isotherm. This indicated that the adsorption occurred in cereal samples was a multilayer adsorption and cereal samples were of a microcapillary structure. In addition, the adsorption in cereal samples decreased as temperature increased. PMID- 10795578 TI - Formulating a new high fiber rusk for production on commercial scale. AB - Three formulas were prepared for production of high dietary fiber rusk on production scale. The preparation was based on partial replacement of wheat flour with different levels of wheat bran, barley and maize flours. Chemical composition, dough characteristics, baking performance and sensory evaluation of rusk were investigated. A remarkable improvement in minerals (calcium and phosphorus) and dietary fiber was achieved. Formulas 'A' and 'B' contained about 3 folds dietary fiber and formula 'C' contained about 4 folds compared with control rusk. The rheological properties of the dough were affected by the fiber incorporation in all suggested formulas. Baking performance showed that all rusk formulas were lower in loaf volume and higher in loaf weight than control sample. Physical measurements and sensory characteristics of rusk indicated generally that all formulas were acceptable, but formula B which contains 70 g wheat flour +10 g wheat bran +10 g maize flour +10 g barley flour seems to be generally superior in symmetry of shape, crust and crumb colour, crumb texture, break and shred, aroma and taste. PMID- 10795579 TI - Effect of proteins, phytates, ascorbic acid and citric acid on dialysability of calcium, iron, zinc and copper in soy-based infant formulas. AB - The possible effect of ascorbic acid, citric acid, proteins and phytate on dialysability of Ca, Fe, Zn and Cu in soy-based infant formulas is studied, taking dialysability as a measure of the amount of element available for absorption. Different dialysis percentages for similar element contents in different formulas are found. A regression analysis was applied between Ca, Zn, Cu and Fe dialysis percentages and soy-based formula components to estimate the possible influence of the latter on the dialysability of the elements. Significant correlations were found between citric acid contents and dialysability of Zn and Fe. No correlations were found between protein, ascorbic acid and phytic acid contents and the dialysis percentages of the four minerals. However, we must to point out that the range of protein contents was narrow and the ascorbic acid: iron ratio was high in our formulas. PMID- 10795580 TI - The effects of electrical stimulation on selected biochemical and histological characteristics of different mutton muscles. AB - The effects of electrical stimulation on selected biochemical characteristics and histological structure of the mutton Longissimus dorsi (LD) and Semimembranosus (SM) muscles were studied. Electrical stimulation had significant influence on the histological characteristics such as sarcolemma disruption, nuclear disorganization, contracture banding and cellular tearing. However, sarcomere length, total and free beta-glucuronidase activities as well as collagen characteristics were not significantly affected by electrical stimulation. Different muscles did not influence histological parameters and beta glucuronidase activities. PMID- 10795581 TI - Determination of cholesterol oxidation products in milk powder and infant formulas by gas chromatography and mass spectrometry. AB - In this paper a method for the cholesterol oxidation products (oxysterols) determination in milk powder and infant formulas has been presented. In the sample preparation step lipids transesterification has been performed. The recoveries of oxysterols have been determined and ranged from 94.2% to 99.9% for all but 20a-hydroxy cholesterol (74.2%). Detection limits were 0.018-0.034 ppm and the relative standard deviations (RSD) values were 4.6%-18.3%. The method has been utilized for the determination of oxysterols in milk-based infant formulas and milk powder available on the market. The concentration of oxysterols was between 0.04 and 4.20 ppm of a lipid extract fraction. PMID- 10795582 TI - Properties and stability of butter oil obtained from milk and yoghurt. AB - The physical and chemical characteristics and thermal stability of butter oil produced from cow's milk by two different methods were studied. Butter oil samples from cow's milk were made (i) directly from milk and (ii) from yoghurt. Samples were autoxidized at 60, 70 and 80 degrees C in the dark and the reaction monitored by peroxide, thiobarbituric acid and free fatty acid values. Peroxide and thiobarbituric acid values increased as the temperature increased. The increase of the acid value was not significant. The thermal stability was highest in butter oil produced from yoghurt. Oxidative changes compared to hydrolytic changes are of greater significance in the thermal stability of butter oil samples. Arrhenius parameters and activation energies were calculated for the peroxide value data. The percent loss of linolenic acid was about 3 times faster than that of linoleic acid. PMID- 10795583 TI - Assessment of oleoresin and gingerol contents in gamma irradiated ginger rhizomes. AB - Oleoresin and gingerol contents in gamma irradiated dried ginger rhizomes were evaluated to determine the effect of radiation and storage on these constituents of ginger. Dried ginger rhizomes were subjected to 0, 5 and 10 kilogray (kGy) doses of gamma rays from 60Co source. The oleoresin and gingerol contents were monitored for 9 months. Radiation treatment (10 kGy) reduced the decrease of the oleoresin content of ginger during the storage period by 14% in unground samples and 11% in ground samples. There was a dose-dependent decrease in the 6-gingerol content of the ground ginger decreased by 65.6, 67.4 and 70.4% for the 0, 5, and 10 kGy samples, respectively, while the corresponding values for the ungrounded ginger samples were 37.8, 40.0 and 44.3% at the end of the storage period. PMID- 10795584 TI - Four-week feeding studies with differently processed dehulled faba bean (Vicia faba L. minor) products in rats. PMID- 10795585 TI - Investigation of the bioavailability of zinc and calcium from some tropical tubers. PMID- 10795586 TI - Biosynthesis of glucuronic acid by means of tea fungus. PMID- 10795587 TI - Blood cultures: clinical aspects and controversies. AB - During the past two decades, there have been important advances in blood culture methodology. These advances have resulted in earlier detection and identification of pathogens causing bloodstream infections. However, there are many facets of the blood culture as a diagnostic test that are not affected by new culture methods or systems that continue to cause problems with interpretation of results. The objective of this review is to focus on those factors influencing the results of blood cultures that have clinical relevance. Such factors include skin preparation, timing, procurement techniques, volume of blood obtained, number of cultures, anaerobic blood cultures, and contamination. In addition, bacteremia prediction models are discussed and suggestions are provided as to how these models could be of greater clinical use. Blood culture methods and systems are not discussed in this review. PMID- 10795588 TI - Serological methods for diagnosis of Helicobacter pylori infection and monitoring of eradication therapy. AB - Several methods can be used to diagnose Helicobacter pylori infection. Invasive methods include detection of the bacterium in gastric biopsy specimens by culture, immunohistochemistry, rapid urease tests, or the polymerase chain reaction. Noninvasive or less invasive detection methods include the urea breath test and serological methods. The urea breath test is based on the detection of 13CO2 or 14CO2 in breath, produced by bacterial urease in the stomach after labelled urea is swallowed. Serological methods are based on the detection of Helicobacter pylori-specific antibodies in serum, saliva, or urine. In this review, the performance and diagnostic value of several serological methods, such as enzyme immunoassay, rapid office-based assays, and Western blot, will be discussed in relation to biopsy-based methods and the urea breath test. In addition, the value of serological assays for monitoring eradication of Helicobacter pylori infection following treatment will be discussed. The diagnostic performance of properly evaluated serological assays is comparable to that of biopsy-based methods and the urea breath test. To monitor eradication of Helicobacter pylori infection following therapy, quantitative enzyme immunoassays can be used, especially in patients with high pretreatment antibody titres. PMID- 10795589 TI - Epidemiological typing of Bordetella pertussis isolates: recommendations for a standard methodology. AB - Pertussis is re-emerging in vaccinated populations, and to gain insight into the reasons for this development population-based studies are necessary. Unfortunately, various techniques are used to study Bordetella pertussis populations, hampering comparison between studies. A standard methodology for epidemiological typing of Bordetella pertussis isolates is proposed which is based on serotyping, pulsed-field gel electrophoresis and gene typing. Such a standard approach will allow comparisons between studies performed in different laboratories. Comparisons may reveal whether the epidemiological differences observed between countries are due for instance to different Bordetella pertussis populations or different vaccines used. PMID- 10795590 TI - Molecular evidence of nosocomial transmission of hepatitis C virus in a haemodialysis unit. AB - The molecular epidemiology of type 2a hepatitis C virus (HCV) infections in patients undergoing haemodialysis in the same unit in a Turkish hospital was investigated. Of nine HCV-infected patients four were infected with type 2a, four with type 1b and one with type 1a viruses. Since type 2 HCV infections in the Turkish population are rare, the possibility of nosocomial infection was investigated by means of phylogenetic analysis of viral sequences amplified by the polymerase chain reaction in the NS5b region. One of the samples failed to show amplification and therefore could not be sequenced. The sequences of the remaining three virus samples were grouped closely in a cluster within the type 2a group. The results thus showed that three patients were infected with the same HCV type 2a strain. Seroconversion and clinical data suggested that these patients may have been infected on different occasions, there being possibly more than one mode of transmission. Breaches in infection control procedures and lack of environmental decontamination between two haemodialysis sessions were probably the causes of HCV infections in these patients. PMID- 10795591 TI - Evaluation of the inactivation of infectious Herpes simplex virus by host-defense peptides. AB - A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide microplate assay was adapted to screen for the ability of 20 host-defense peptides to inactivate herpes simplex virus type 1 and type 2. The procedure required minimal amounts of material, was reproducible, and was confirmed with standard antiviral testing techniques. In screening tests, with the exception of melittin, a highly cytotoxic and hemolytic peptide found in bee venom, the alpha-helical peptides in our test panel (magainins, cecropins, clavanins, and LL-37) caused little viral inactivation. Several beta-sheet peptides (defensins, tachyplesin, and protegrins) inactivated one or both viruses, sometimes with remarkable selectivity. Two peptides were identified as having antiviral activity against both viruses, indolicidin (a tryptophan-rich peptide from bovine neutrophils) and brevinin-1 (a peptide found in frog skin). The antiviral activity of these two peptides was confirmed with standard antiviral assays. Interestingly, the antiviral activity of brevinin-1 was maintained after reduction and carboxamidomethylation, procedures that abolished its otherwise prominent hemolytic and cytotoxic effects. PMID- 10795592 TI - Booster effect of low doses of tetanus toxoid in elderly vaccinees. AB - In order to improve the immunity to diphtheria, the recommended booster dose of diphtheria/tetanus vaccine for adults in Sweden was changed in 1986 from 0.5 ml of tetanus vaccine with a small diphtheria dose to 0.25 ml of a diphtheria/tetanus vaccine containing 7.5 Lf tetanus toxoid and 30 Lf diphtheria toxoid/ml. This change resulted in an increase in the dose of diphtheria toxoid from 0.5 Lf to 7.5 Lf, but a decrease in the recommended booster dose of tetanus toxoid from 3.75 Lf to 1.9 Lf. The aim of the present study was to investigate whether this lower dose of tetanus toxoid was also sufficiently protective for elderly people. Two hundred adults (median age 76 years, range 60-92 years) with no history of tetanus vaccination during the past 10 years volunteered for the study. One hundred two vaccinees were inoculated with 1.9 Lf tetanus toxoid (0.25 ml) and 98 with 3.75 Lf tetanus toxoid (0.5 ml). Paired serum samples were analysed by the toxin-binding inhibition assay. Side effects were few and mild, without significant differences between the groups. Response rates were similar, with the 3.75 Lf dose eliciting a marginally higher antitoxin response. The prevaccination geometric mean titre was the same for both groups: 0.03 IU/ml. Postvaccination geometric mean titres were 1.18 IU/ml for the 3.75 Lf group and 1.93 IU/ml for the 7.5 Lf group, respectively (difference not significant). Forty seven percent of the vaccinees had a prevaccination titre of < or =0.01 IU/ml. Postvaccination, 85% had a titre >0.01 IU/ml. Booster vaccination with tetanus vaccine containing only 1.9 Lf of tetanus toxoid was thus found to induce an excellent immune response in elderly people, with few side effects resulting. PMID- 10795593 TI - Tuberculin skin reactivity in Italian military recruits tested in 1996-1997. AB - In 1996-1997 data was collected and a Mantoux tuberculin test performed in 2882 Italian military recruits aged 18-23 years in order to establish the prevalence of tuberculin reactivity. In addition, the annual risk of infection, defined as the probability that a non-infected individual would be infected during the following year, was calculated. Of the 2882 recruits, 513 had received a BCG vaccination, the remaining 2369 had not. The overall prevalence of subjects with a tuberculin skin reaction size >5 mm (the cut-off point for positivity corresponding to the antimode in the reaction size frequency curve) was 6.1% (144/2369). The prevalence of skin reactivity increased with age but remained similar when related to area of residence, duration of father's school education and family size. The same general trend was observed if the standard pre established cut-off point of 10 mm was used. In this case the overall prevalence of a positive skin reaction was 4% (95/2369). The annual risk of infection was 0.3% for a prevalence of tuberculin skin reactivity of 6.1% (cut-off point 5 mm) and 0.19% for a prevalence of 4% (cut-off point 10 mm). Analysis of the population sample vaccinated with BCG showed a lack of correlation between the positive reaction after vaccination reported retrospectively by the subject and the current skin reaction observed by the physician in this study (K = 0.254). Moreover, a significant difference was observed between the skin reaction in subjects vaccinated with BCG in 1993-1994 (average size 12.5 mm) and that of subjects vaccinated in 1995-1996 (average size 10.1 mm, P<0.01), probably as a consequence of mycobacteria circulating in the general population which act as a natural booster in people already vaccinated with BCG. A booster effect of tuberculin in Mantoux assays also cannot be excluded. PMID- 10795594 TI - Infection caused by Nocardia farcinica: case report and review. AB - Nocardia farcinica is a rare Nocardia species causing localised and disseminated infections. A case of Nocardia farcinica infection is presented, and 52 cases previously reported in the literature are reviewed. The hosts usually had predisposing conditions (85%), and acquired the infection through the respiratory tract or skin; the infection then often spread to the brain, kidney, joints, bones and eyes. Pulmonary or pleural infections (43%), brain abscesses (30%) and wound infections (15%) which failed to respond to conventional antimicrobial therapy were the more frequent forms of infection. Nocardia farcinica was frequently isolated from pus (100% of samples), bronchial secretions (41%) and biopsy specimens (63%), but isolation from blood and urine, as in the case presented here, is rare. Antibiotic therapy was adequate in 61% of the patients in whom it was specified, the agents most frequently given being trimethoprim sulfamethoxazole (54%), amikacin combined with imipenem (7%) and amoxicillin clavulanate (7%). The high mortality (31%) can be attributed to the severe underlying diseases present, difficulties encountered in identifying the pathogen, inappropriate therapy and late initiation of therapy. Although an infrequent pathogen, Nocardia farcinica should be kept in mind as a cause of infection especially in immunosuppressed patients with indolent infections not responding to third-generation cephalosporins. PMID- 10795595 TI - Effect of antiretroviral therapy on cryptosporidiosis and microsporidiosis in patients infected with human immunodeficiency virus type 1. AB - To better understand whether potent antiretroviral therapies can modify the natural history of HIV-1-associated microsporidiosis and cryptosporidiosis, the response to antimicrobial treatment of these opportunistic infections was evaluated in patients with or without antiretroviral treatment. Fifty patients with diarrhoea, all positive for Cryptosporidium parvum or Enterocytozoon bieneusi, were included in the study. Retrospective data were collected concerning demographics, clinical and microbiological characteristics of the parasitic infection, antiretroviral therapy and prophylaxis against opportunistic infections. Faecal samples were prepared using the Richie formalin-ethyl acetate method and stained using the modified Ziehl-Neelsen method for detection of Cryptosporidium parvum and Isospora belli, the modified trichrome and calcofluor white technique for detection of Enterocytozoon spp., and iodine for detection of ova, cysts or vegetative forms. Diarrhoea was defined as an abnormal increase in stool liquidity, an abnormal increase in stool frequency and a daily stool weight of more than 250 g for a period of at least 4 days. Patients treated with double antiretroviral therapy or protease inhibitors demonstrated an excellent response and a sustained therapeutic effect after follow-up (range, 5-36 months). The relapse of cryptosporidiosis in two patients who discontinued antiretroviral therapy suggests that the infection might remain in a latent stage. The resolution of the diarrhoea seems to be related to an increased CD4+ cell count rather than to the viral load. In conclusion, these data strongly support the hypothesis that combination antiretroviral therapy is able to greatly modify the course of cryptosporidiosis and microsporidiosis in patients infected with HIV-1. PMID- 10795596 TI - Report on five cases of tularaemic pneumonia in a tularaemia outbreak in Spain. AB - A report is given on five cases of atypical tularaemic pneumonia selected from among 140 cases of tularaemic infection in a previously reported outbreak occurring in 1997. Prior to this outbreak no human cases of tularaemia had been reported in Spain. All cases were diagnosed serologically. All five patients reported on here had a mild form of the disease, which was treated successfully with streptomycin in four cases and ciprofloxacin in one case. Tularaemic pneumonia should be considered in the differential diagnosis of atypical pneumonia in Spain, especially in hunters and other persons who handle animal carcasses. PMID- 10795597 TI - A case of Enterocytozoon bieneusi infection in an HIV-negative renal transplant recipient. AB - Reported here is a case of microsporidiosis that occurred in an HIV-negative renal transplant recipient. The patient developed protracted diarrhea 18 months following transplant surgery. Many spores of Enterocytozoon bieneusi were detected in stool smears using a modified trichrome staining method. Identification was confirmed using the polymerase chain reaction. Histological examination of duodenal biopsies revealed numerous spores in the cytoplasm of enterocytes. Tacrolimus and steroid regimens were decreased, treatment with mycophenolate mofetil was discontinued, and the patient was given albendazole and metronidazole for 2 weeks. The diarrhea resolved after 15 days of treatment; 2 months later the patient had recovered completely. A more systematic search for microsporidia using specific staining procedures should be performed in transplant recipients who develop severe diarrhea. PMID- 10795598 TI - Unilateral vocal cord paralysis as a result of a Nocardia farcinica laryngeal abscess. AB - Nocardia farcinica is an emerging pathogen in immunocompromised hosts, accounting for 20% of Nocardia isolates in the USA and 13-44% of isolates in Europe. The case of a 72-year-old lymphoma patient with a laryngeal abscess caused by Nocardia farcinica is presented. The initial clinical manifestation was unilateral vocal cord paralysis, which improved following surgical drainage of the abscess and therapy with imipenem. The English-language literature on human Nocardia farcinica infection is reviewed. PMID- 10795599 TI - Comparative activity of moxifloxacin in vitro against obligately anaerobic bacteria. AB - The antimicrobial activity of moxifloxacin and seven other antibiotics (four of them quinolones) against 292 strains of obligately anaerobic bacteria was assessed employing a broth microdilution technique performed in Wilkens-Chalgren broth. MIC50/MIC90 values (mg/l) for moxifloxacin were as follows: Bacteroides fragilis (n = 62) 0.25/2, Bacteroides ovatus (n = 70) 1/4, Bacteroides vulgatus (n = 29) 0.25/1, Bacteroides thetaiotaomicron (n = 17) 2/2, Bacteroides caccae (n = 11) 1/2, Prevotella spp. (n = 11) 0.25/2, Fusobacterium spp. (n = 17) 1/4, Bilophila wadsworthia (n = 29) 0.5/1, and Clostridium spp. (n = 29) 0.125/0.5, respectively. MIC50 values (mg/l) for Bacteroides distasonis (n = 8) and Peptostreptococcus spp. (n = 9) were 0.25. The results indicated that moxifloxacin was almost as active as trovafloxacin, as active as gatifloxacin, and more active than levofloxacin and ciprofloxacin against the anaerobes tested. PMID- 10795600 TI - Evaluation of an enzyme-linked viral inducible system for the rapid detection of Herpes simplex virus. AB - A commercial enzyme-linked viral inducible system (ELVIS HSV; BioWhittaker, USA) was evaluated in comparison with the spin-amplified tube cell culture (SATCC) method for the rapid detection of herpes simplex virus in 1007 clinical specimens. A total of 91 (9%) specimens were positive by SATCC. The sensitivity, specificity, positive predictive value, and negative predictive value of ELVIS was 88%, >99%, 99%, and 99%, respectively. Herpes simplex virus was detected sooner by ELVIS than by SATCC in 34 of 80 (42%) specimens. Preincubated ELVIS shell vials held at room temperature for 24 h prior to reincubation and inoculation produced results similar to those of freshly preincubated shell vials, with no reduction in either the number or the staining intensity of the infected cells. The results of this study indicate that ELVIS HSV is an accurate method for the rapid detection of herpes simplex virus in a wide variety of clinical specimens. PMID- 10795601 TI - Clinicoepidemiological features of immigrants with tuberculosis living in Madrid, Spain. PMID- 10795602 TI - Influence of prior HIV-1 infection on the development of chronic hepatitis B infection. PMID- 10795603 TI - Comparison of an enzyme immunoassay versus a rapid latex test for serodiagnosis of Helicobacter pylori infection. PMID- 10795604 TI - Posttraumatic stress disorder: epidemiology and health-related considerations. AB - Epidemiologic studies show that prevalence of trauma and posttraumatic stress disorder (PTSD) is substantial in modern society. Most people will experience a traumatic event at some point in their life, and up to 25% of them will develop the disorder. Demographic and socioeconomic factors also play a role in the risk for exposure to traumatic experiences and subsequent PTSD. Psychiatric history, both personal or in family members, increases the likelihood of being exposed to trauma and of developing PTSD once exposed. Traumatic exposure and PTSD have an impact on the individual's health, health care service utilization, and general functioning. Such effects provoke a considerable economic burden not only for those persons experiencing trauma and PTSD, but also for the health care system and society as a whole. PMID- 10795605 TI - Biology of posttraumatic stress disorder. AB - An understanding of the biological basis of posttraumatic stress disorder (PTSD) requires an examination of the underlying neurobiology of fear and the factors that might contribute to an unsuccessful termination of the fear response in some individuals. Several factors may lead to an inadequate termination of a stress response, and the failure to contain the biological alterations initiated by stress may have long-term adverse consequences. In particular, a prolonged continuation of biological responses following stress may lead to an inappropriate pairing of the traumatic memory with distress and may then initiate a cascade of secondary biological alterations. This article examines some of the biological alterations in PTSD and develops a framework for understanding the development progression of the neurobiology of this disorder. PMID- 10795606 TI - Comorbidity of psychiatric disorders and posttraumatic stress disorder. AB - Posttraumatic stress disorder (PTSD) commonly co-occurs with other psychiatric disorders. Data from epidemiologic surveys indicate that the vast majority of individuals with PTSD meet criteria for at least one other psychiatric disorder, and a substantial percentage have 3 or more other psychiatric diagnoses. A number of different hypothetical constructs have been posited to explain this high comorbidity; for example, the self-medication hypothesis has often been applied to understand the relationship between PTSD and substance use disorders. There is a substantial amount of symptom overlap between PTSD and a number of other psychiatric diagnoses, particularly major depressive disorder. It has been suggested that high rates of comorbidity may be simply an epiphenomenon of the diagnostic criteria used. In any case, this high degree of symptom overlap can contribute to diagnostic confusion and, in particular, to the underdiagnosis of PTSD when trauma histories are not specifically obtained. The most common comorbid diagnoses are depressive disorders, substance use disorders, and other anxiety disorders. The comorbidity of PTSD and depressive disorders is of particular interest. Across a number of studies, these are the disorders most likely to co-occur with PTSD. It is also clear that depressive disorder can be a common and independent sequela of exposure to trauma and having a previous depressive disorder is a risk factor for the development of PTSD once exposure to a trauma occurs. The comorbidity of PTSD with substance use disorders is complex because while a substance use disorder may often develop as an attempt to self medicate the painful symptoms of PTSD, withdrawal states exaggerate these symptoms. Appropriate treatment of PTSD in substance abusers is a controversial issue because of the belief that addressing issues related to the trauma in early recovery can precipitate relapse. In conclusion, comorbidity in PTSD is the rule rather than the exception. This area warrants much further study since comorbid conditions may provide a rationale for the subtyping of individuals with PTSD to optimize treatment outcomes. PMID- 10795607 TI - Posttraumatic stress disorder: psychological factors and psychosocial interventions. AB - In this article, we propose that successful processing of traumatic events involves emotional engagement with the trauma memory, organization of the trauma narrative, and correction of dysfunctional cognitions that often follow trauma. We further propose that the success of psychosocial treatments of posttraumatic stress disorder hinges on the ability of the treatments to address impairments in these processes. We focus our presentation of psychosocial interventions on cognitive-behavioral treatments (CBT), since this approach had gained the most empirical support to date, and describe the results of controlled trials that compare the relative efficacy of several CBT interventions. PMID- 10795608 TI - Antidepressant treatment of posttraumatic stress disorder. AB - Recent large double-blind, placebo-controlled trials have indicated that sertraline is an effective and well-tolerated treatment for posttraumatic stress disorder (PTSD). The avoidance/numbing symptom cluster improved the most significantly with sertraline, but significant improvements were also noted for the intrusive/reexperiencing and arousal symptom clusters. Smaller double-blind, placebo-controlled trials have also indicated that fluoxetine is an effective treatment for PTSD. Multiple small, open studies with other selective serotonin reuptake inhibitors and newer antidepressants indicate that these medications show some promise. Older studies indicate some efficacy for tricyclic antidepressants, and monoamine oxidase inhibitors are a reasonable choice, particularly for intrusive/reexperiencing symptoms. PMID- 10795609 TI - What might the psychobiology of posttraumatic stress disorder teach us about future approaches to pharmacotherapy? AB - This review considers future directions for developing effective drugs for posttraumatic stress disorder (PTSD). At present, we have embarked upon an empirical approach in which pharmacologic research consists of clinical trials with agents, such as antidepressants, anxiolytics, and anticonvulsants, initially developed for different purposes. The approach taken here is theoretical rather than empirical, starting with what is known about the unique pathophysiology of PTSD and then predicting the types of pharmacologic agents that might prove effective in the future. Such classes of compounds include corticotropin releasing factor antagonists, neuropeptide Y enhancers, antiadrenergic compounds, drugs to down-regulate glucocorticoid receptors, more specific serotonergic agents, agents normalizing opioid function, substance P antagonists, N-methyl-D aspartate facilitators, and antikindling/antisensitization anticonvulsants. PMID- 10795610 TI - Coding for percutaneous nephrolithotomy and related procedures. PMID- 10795611 TI - Controversial cases in endourology. PMID- 10795612 TI - Extracorporeal shockwave lithotripsy of 2000 urinary calculi with the modulith SL 20: success and failure according to size and location of stones. AB - PURPOSE: We analyzed the successes and failures of SWL in the treatment of 2016 urinary calculi stratified according to size and position in the urinary tract. METHODS: All the patients were treated with a Modulith SL-20 (Storz Medical). RESULTS: The overall stone-free rate, regardless of the size and position of the stone, was 87.4%. The rate was different for kidney and ureteral stones. The stone-free rate observed for the different positions of the calculi within the kidney was upper calix 89.2%, middle calix 90.5% lower calix 84.8%, and renal pelvis 86.0%. For staghorn calculi, the stone-free rate was 60.0%. The stone-free rate for the different positions of calculi within the ureter was: upper ureter 84.3%, iliac ureter 82.4%, and pelvic ureter 91.0%. For calculi >24 mm, the retreatment rate increased, and the success rate dropped sharply. CONCLUSION: Extracorporal shockwave lithotripsy should be the first therapeutic option for urinary calculi of <24 mm regardless of their position in the urinary tract. PMID- 10795613 TI - Laparoscopic nephropexy: long-term follow-up--Washington University experience. AB - BACKGROUND AND PURPOSE: Recently, laparoscopy has been reported as a minimally invasive approach for performing nephropexy in patients with symptomatic nephroptosis. Herein, we report our long-term follow-up of patients undergoing laparoscopic nephropexy for this indication. PATIENTS AND METHODS: Fourteen women presenting with right flank pain and radiologically documented nephroptosis underwent transperitoneal laparoscopic nephropexy. The hospital data were evaluated for operative time, time to oral intake, time to ambulation, amount of parenteral analgesics, and hospital stay. Pain analog scores and postoperative questionnaires were used to assess the long-term postoperative recovery of the patients. RESULTS: The average operative time was 4.1 hours (range 2.5-6.5 hours). The patients resumed oral intake an average of 16.5 hours (range 15-48 hours) postoperatively. Analgesic requirements averaged 37 mg of morphine sulfate equivalent (range 15-80 mg of morphine equivalent). The average hospital stay was 2.6 days (range 2-5 days). The average follow-up time for the 14 patients was 3.3 years, with an average 80% improvement in their pain (range 56%-100%). On average, the patients resumed their usual activities 6 weeks postoperatively (range 1-12 weeks). CONCLUSION: Nephropexy can be safely and effectively accomplished laparoscopically, with durable radiographic and clinical resolution of the signs and symptoms. PMID- 10795614 TI - Combined antegrade and retrograde endoscopic approach for the management of urinary diversion-associated pathology. AB - BACKGROUND: Endourologic management of stones and strictures in patients with a urinary diversion is often cumbersome because of the absence of standard anatomic landmarks. We report on our technique of minimally invasive management of urinary diversion-associated pathology by means of a combined antegrade and retrograde approach. PATIENTS AND METHODS: Five patients with urinary diversion-associated pathology were treated at our institution between May 1997 and October 1998. Their problems were: an obstructing ureteral stone in a man with ureterosigmoidostomy performed for bladder extrophy; two men with a valve stricture in their hemiKock urinary diversions; an anastomotic stricture in a man with an ileal loop diversion; and a long left ureteroenteric stricture in a man with a right colon pouch diversion. After percutaneous placement of an guidewire across the area of interest, the targeted pathology was accessed via a retrograde approach using standard semirigid or flexible fiberoptic endoscopes. Postoperative follow-up with intravenous urography, differential renal scan, or both was performed at 3 to 24 months (mean 12 months). RESULTS: The combined antegrade and retrograde approach allowed successful access to pathologic areas in all patients. Holmium laser/Acucise incision of stenotic segments or ballistic fragmentation of stones was achieved in all cases without perioperative complications. None of the strictures with an initially successful outcome has recurred; however, in one patient, the procedure failed as soon as the internal stent was removed. The patient with the ureteral calculus remains stone free, and his ureterosigmoidostomy is patent without evidence of obstruction on his last imaging study, 24 months postoperatively. CONCLUSIONS: Combined antegrade and retrograde endoscopic access to the area of interest is our preferred method of approaching pathologic problems in patients with a urinary diversion. An antegrade nephrostogram provides better delineation of anatomy, while through-and through access enables rapid and easier identification of stenotic segments that may be hidden by mucosal folds. Furthermore, this approach allows the use of larger semirigid or flexible endoscopes in conjunction with more efficient fragmentation devices, resulting in enhanced vision from better irrigation. Finally, an initial endoscopic approach may be preferred because its failure does not compromise the success of future open surgery. PMID- 10795615 TI - Percutaneous treatment of large symptomatic renal cysts. AB - PURPOSE: To evaluate the efficacy of alcohol in combination with tetracycline for the treatment of symptomatic renal cysts. PATIENTS AND METHODS: Twenty-four patients age 45 to 77 years (mean 66 years) with a large (5-13-cm; mean 7.5-cm) symptomatic renal cyst associated with flank pain were treated by aspiration under ultrasound guidance and injection of alcohol and tetracycline. Patients were followed with ultrasonography at 1, 6, and 12 months and once a year thereafter. RESULTS: The aspirated volume ranged from 65 to 1500 mL (mean 360.5 mL). Two patients experienced mild pain during alcohol injection, but the procedure was completed successfully. One patient reported severe pain after tetracycline injection. The tetracycline was immediately aspirated, and the procedure was then aborted. The remaining patients were relieved of their symptoms after treatment, and they remained symptom free during a mean follow-up of 20 months (range 7-36 months). CONCLUSIONS: The combination of alcohol and tetracycline is safe and effective and offers a very favorable minimally invasive therapeutic alternative for the treatment of symptomatic renal cysts. PMID- 10795616 TI - Endoureterotomy for congenital primary obstructive megaureter: preliminary report. AB - BACKGROUND AND PURPOSE: Congenital obstructive megaureter (COMU) may be unilateral or bilateral and may present in later years of life. If the obstruction is not relieved in good time, deleterious effects on the proximal ureter and kidney are well known. Stones may complicate the situation further. So far, the only treatment that has been available is to disconnect the ureter proximal to the site of obstruction, remove any stone, and reimplant the ureter into the bladder after any necessary tailoring. After noticing the encouraging results of endopyelotomy for congenital ureteropelvic junction obstruction, we decided to use the same principle for the management of COMU, as similar functional pathology is present in both situations. PATIENTS AND METHODS: Our technique was applied in six ureters in five adult patients. After cystoscopic evaluation of the bladder and ureteral orifice, a guidewire was advanced up the ureter, and the lower ureter was dilated. A ureteroscope or other suitable endoscopic instrument was passed, and the obstructed segment of the ureter was incised at the 6 o'clock position with pure cutting current. All of the layers of the ureter were incised in the long axis through the entire obstructive segment, so as to expose the periureteral areolar tissue. If necessary, a similar cut was made at the 12 o'clock position. Utmost care was taken not to incise the bladder mucosa. A double-J stent was inserted for 3 weeks. RESULTS: Follow-up for 1 to 4 years showed free drainage of urine into the bladder, with marked reduction in proximal stasis and freedom from recurrent infection and pain. CONCLUSION: Although our series is small and follow- up is relatively short, it appears that endoureterotomy is a safe and effective treatment for COMU. PMID- 10795617 TI - Mini-percutaneous nephrolithotomy. AB - Standard percutaneous nephrolithotomy is highly effective for the removal of renal calculi. However, significant morbidity has been associated with this procedure. Consequently, many urologists inappropriately defer to a less effective procedure to reduce patient morbidity. This practice may increase the total number of procedures needed for treatment and result in a substantial increase in health care costs. Mini-percutaneous nephrolithotomy using a 13F ureteroscopy sheath is described to reduce the morbidity associated with standard percutaneous nephrolithotomy while maintaining its efficiency and effectiveness for stone removal. The indications and technique for mini-percutaneous nephrolithotomy and our results are summarized. PMID- 10795618 TI - Ureteroscopic approach to upper-tract urothelial tumors. AB - Transitional-cell carcinoma (TCC) of the upper urinary tract has traditionally been managed by nephroureterectomy, whereas nephron-sparing surgery has been reserved for those few patients with solitary kidneys or bilateral lesions. However, with the introduction of improved diagnostic and therapeutic technology, including smaller ureteroscopes and working instruments, and the concomitant ease of surveillance, ureteroscopic treatment of upper-tract urothelial tumors has become a reasonable alternative to open operative intervention in patients requiring conservative management. Furthermore, as preoperative grading and staging have improved, ureteroscopic treatment of upper-tract urothelial tumors is assuming an increasingly important role in the management of some patients who might have otherwise been treated with a nephroureterectomy. The technique of ureteroscopic resection is described in detail. PMID- 10795619 TI - Treatment of urinary calculi with ureteroscopy and Swiss lithoclast pneumatic lithotripter: report of 150 cases. AB - PURPOSE: To present our clinic experience with the Swiss Lithoclast pneumatic lithotripter in the endoscopic management of urinary calculi. PATIENTS AND METHODS: From August 1994 to December 1997, 145 patients with ureteral calculi and 5 patients with urethral calculi were treated with the Swiss Lithoclast. RESULTS: In the ureteral stone group, ureteroscopic addressing of the stones was successful in 133 patients. In 27 patients, the stones were partially fragmented and remained in situ or were pushed back to the calices. They were subsequently treated successfully with SWL. Stones were fragmented in a single session in 101 cases. Complications associated with the procedure included five perforations and four urinary tract infections. All of the five urethral stone patients were treated successfully with pneumatic lithotripsy. The overall successful fragmentation rate thus was 70.7% (106 of 150) and 88.7% (133 of 150) in combination with adjuvant SWL. CONCLUSIONS: We have found Swiss Lithoclast pneumatic lithotripsy to be a safe, effective, and economical treatment method for urinary calculi. If combined with other modalities such as SWL, this treatment will be even more effective. PMID- 10795620 TI - Bilateral single-session percutaneous nephrolithotomy: a feasible and safe treatment. AB - BACKGROUND AND PURPOSE: The treatment of bilateral urinary calculus disease is often staged, irrespective of the modality of the treatment. Bilateral simultaneous percutaneous nephrolithotomy (PCNL) is still considered by many to be adventurous and risky. We carried out this prospective study of bilateral PCNL under a single anesthesia to study the feasibility, success rate, and complications if any. PATIENTS AND METHOD: From September 1996 to May 1999, 25 consecutive medically fit patients with bilateral renal calculi needing PCNL were subjected to bilateral PCNL under a single anesthesia. RESULTS: The PCNL could be accomplished bilaterally in 24 patients (96%). In one patient, the second-side PCNL was abandoned because of excessive bleeding on the first side. A total of 58 tracts and 27 sessions were required for complete treatment of the 48 renal units in the 24 successfully treated patients. The average operating time required for the procedure was 122 minutes. Nineteen patients were rendered stone free in one session, and two more patients were made stone free in a second session. Four renal units in three patients with residual calculi were cleared with double-J stenting and SWL. CONCLUSION: Bilateral PCNL in a single session is feasible and safe and can be carried out without increased morbidity. We advocate that an endourologist be prepared for bilateral PCNL in the patients in whom it is indicated. The opposite-side PCNL can be done if the first-side treatment is accomplished smoothly and in a reasonable time. PMID- 10795621 TI - Percutaneous nephrolithotripsy in ectopic kidneys. AB - BACKGROUND: Percutaneous nephrolithotomy (PCNL), although an accepted treatment modality in anatomically normal kidneys, is still not universally performed for calculi in pelvic ectopic kidneys. Fear of injury to abdominal viscera makes it a technically challenging procedure. PATIENTS AND METHODS: We have performed PCNL in nine patients with calculi in pelvic ectopic kidneys. Technical factors which made this procedure safe include ultrasound-guided puncture, use of a mature tract or an Amplatz sheath, routine postoperative double-J stenting, and nephrostogram prior to nephrostomy tube removal. RESULTS: Complete stone clearance was achieved in all cases. Six patients were treated in a single stage, while three patients required two stages. Seven patients needed only one tract, and two needed two tracts. No notable complications were encountered. The average hospital stay was 5.2 days. CONCLUSION: With proper precautions and meticulous technique, PCNL is a safe and effective modality to treat calculi in pelvic ectopic kidney. PMID- 10795622 TI - High-intensive focused ultrasound in localized prostate cancer. AB - BACKGROUND AND PURPOSE: The results of the standard treatment for prostate cancer radical prostatectomy-are not entirely satisfactory. A new local therapy, transrectal high-intensive focused ultrasound (HIFU), has been developed. We reviewed our experience with HIFU for palliation of localized prostate cancer. PATIENTS AND METHODS: Our series included 65 men with confirmed prostate cancer without detectable metastases who were not suitable candidates for radical prostatectomy. After prophylactic suprapubic cystostomy, the patients were treated using the Ablatherm version 2.32 under spinal anesthesia. The effects were monitored by serum prostate specific antigen assays, digital rectal examination, and biopsy. The mean follow-up is 10 months (range 1-18 months). RESULTS: There were no intraoperative or postoperative deaths, and there have been no deaths from prostate cancer. Residual cancer was detected in 35% of the patients in whom only biopsy-positive portions of the prostate were treated and 17% of those in whom the entire gland was treated. Retreatment was performed 1 month after the first session in these patients. The prostate volume increased an average of 30% after treatment, but by 3 months, the gland was 10% to 20% smaller than its original size. Three patients suffered complications secondary to overheating of the rectal wall or treatment too close to the external urethral sphincter. CONCLUSION: The low morbidity, minimal invasiveness, avoidance of systemic side effects, and potentially curative effect make HIFU a potentially useful option for the treatment of localized prostate cancer. PMID- 10795624 TI - Literature watch. PMID- 10795625 TI - Dendritic cell-based vaccines: from mouse models to clinical cancer immunotherapy. AB - B and T lymphocytes are the effectors of specific immunity. However, their function is critically dependent on dendritic cells (DC). DC are professional antigen presenting cells that both initiate and modulate the immune response. The recent breakthrough in the generation of DC from their progenitors has stimulated research on DC in both fundamental and clinical immunology. Objective immune response induction has now been reported in clinical studies using DC. In this review we discuss the development and potential of DC-based vaccines to induce antitumor immunity. PMID- 10795623 TI - Holmium laser ablation of recurrent strictures of urethra and bladder neck: preliminary results. AB - BACKGROUND: The management of patients with recurrent urethral strictures represents a challenge for the practicing urologist. PATIENTS AND METHODS: We used holmium:yttrium-aluminum-garnet (Ho:YAG) laser in the management of recurrent urethral strictures in 13 patients. The energy level was set at 1.0 at a frequency of 10 pulses/sec. No treatment complications were observed. The mean preoperative maximum flow rate by uroflowmetric analysis was 3.8 mL/sec. RESULTS: Nine patients (69%) continue to do well with no symptoms at a median follow-up of 27 months with a mean maximum flow rate of 19 mL/sec. Of the four patients in whom treatment failed, three were retreated with the Ho:YAG laser. One of them was managed by insertion of a permanent urethral stent, another continues to do well without any further treatment, and the other is managed with dilation by self-catheterization. One of the four failures underwent open reconstructive urethroplasty after recurrence following his first treatment with the Ho:YAG laser. CONCLUSION: Our preliminary results suggest that Ho:YAG laser ablation of urethral strictures is safe and might be a reasonable alternative endoscopic treatment for recurrent urethral strictures. PMID- 10795626 TI - Genetic alterations in liver carcinogenesis: implications for new preventive and therapeutic strategies. AB - In this review, genetic changes known to occur in human and experimental animal hepatocarcinogenesis are evaluated comparatively, with the aim of identifying genes that could potentially be targets of new preventive and therapeutic strategies, albeit the fact that although a step-by-step analysis of the premalignant stages has been largely accomplished in experimental hepatocarcinogenesis, this goal is still elusive in the case of humans. Overexpression of several of the genes implicated in the MAPK signaling cascade and cell cycle control appears to be most likely responsible for initiated cells acquiring a proliferating phenotype that facilitates the accumulation of structural changes in additional genes, resulting in the generation of autonomously growing preneoplastic and neoplastic lesions. Several gene abnormalities seen in precancerous lesions of rodents also occur in human hepatocellular carcinomas, suggesting that at least some of them could be present also in human precancerous lesions. Furthermore, there are reports that epigenetic events, such as abnormal DNA methylation, may be critical in hepatocarcinogenesis. DNA hypomethylation is an early event, both in human and experimental hepatocarcinogenesis, and its role in the activation of various genes, has been postulated. In recent years, linkage analysis studies have led to the identification of susceptibility/resistance loci that influence the progression stage of hepatocarcinogenesis in mice and rats. The relevance of these findings, though, will depend on the identification of the genes, and on whether in humans there are genes ortholog with rodent's susceptibility/resistance genes. It is proposed that rodent hepatocarcinogenesis represents a promising model for the identification of genes implicated in the early stages of the process, and that many of these genes may represent key targets for the application of gene therapy in the prevention and treatment of liver cancer. PMID- 10795627 TI - Role of the cytoskeletal protein paxillin in oncogenesis. AB - The focal adhesion is an important cellular structure that is involved in cell signaling, cell motility, and oncogenic transformation. Paxillin is a unique adapter protein that is localized to the focal adhesion and is involved in regulating various functions of the focal adhesion. The predicted amino acid structure for paxillin shows at the amino-terminus five LD motifs, a proline-rich domain, several potential phosphorylation sites and four carboxy-terminal LIM domains. Paxillin interacts with cell surface receptors and the actin cytoskeleton and activates several signal transduction pathways that are known to regulate normal cell physiology. Because paxillin is a central protein within the focal adhesion, it is a common target of many different oncoproteins, such as BCR/ABL, v-Src, and E6. In this review we summarize the current knowledge about the structure/function of paxillin and its family members, its role in integrin, cytokine signaling, and oncogenic transformation. PMID- 10795628 TI - Cancer genetics/epigenetics and the X chromosome: possible new links for malignant glioma pathogenesis and immune-based therapies. AB - Human high-grade gliomas (HGGs) are rapidly progressing heterogeneous brain tumors of unknown etiology and there are no effective treatment modalities available. The recent discovery of cancer-specific antigens has opened new doors for specific tumor-targeted treatments using passive and active immunotherapeutic strategies. In particular, SEREX (serological analysis of recombinant cDNA expression libraries) has aided in the discovery of numerous new tumor antigens. These specific tumor antigens are located on chromosome X and are expressed predominantly in the testes among normal organs, and hence termed Cancer/Testis Antigens (CTAs). We found that the vast majority of HGG patients overexpress a receptor for an immune regulatory cytokine, interleukin 13 (IL-13), which differs from the normal tissue physiological receptor. Interestingly, the HGG-associated receptor protein, IL-13R alpha, is expressed solely in the testes and its gene is localized to chromosome X, which mirror the expression pattern and genomic localization of CTAs. There is little evidence for frequent gross structural abnormalities on chromosome X in HGG. Although the mechanism that causes X chromosome-linked CTAs to be aberrantly expressed in tumors is not fully understood, evidence is beginning to point toward the DNA methylation dysregulation that occurs in tumor cells as being implicit in this process and perhaps in the oncogenic process as well. Therefore, further study of the phenomenon of CTAs may bring the dual benefit of better understanding tumorigenesis and providing new molecular tools for better management of HGGs. Also, we propose that the X chromosome may in fact be an important player in HGG oncogenesis. PMID- 10795629 TI - Flowers produce variations in color saturation by arranging petals at oblique and varying angles. AB - Large color variations can be observed across the face of a flower even when individual petals are the same color. We investigated whether these color variations could be explained by a model that incorporates multiple reflections of light between petals and transmissions of light through petals before the light returns to the observer. The three flowers that we selected for the study exhibited large color variations across the face of the intact flower but had no significant observable difference in color saturation across a single petal or between petals when petals were removed from the flower. We used a spectroradiometer to measure the spectrum across the faces of intact flowers and across individual petals. The measured spectra for all of the flowers were consistent with the proposed model. PMID- 10795630 TI - Color perception through atmospheric haze. AB - The radiance of most objects seen at a distance through the atmosphere is dominated by scattered light of a blue hue that should make the landscape appear predominately blue. However, common experience shows that people can see colors at a distance. A possible explanation of this paradox is that the visual system splits the light into a haze layer and the background landscape. A straightforward mathematical description of this splitting explains the results of a color matching study in the Great Smoky Mountains National Park. In this study, hues of objects seen through haze were found to be constant with changes in optical depth while colorfulness decreased exponentially. PMID- 10795631 TI - Spatiotemporal filters in the detection of background modulation targets. AB - The background modulation method has been proposed as a useful test of early visual mechanisms [Biol. Cybern. 37, 77 (1980); Biol. Cybern. 47, 173 (1983)]. The task involves measuring detection thresholds for a luminous spot (increment) drifting over a spatially or temporally modulated background. The study explores the nature of the detecting mechanism in terms of spatial and temporal filters for both spatial and temporal background modulations. In both cases we find that thresholds can be explained by spatial contrast cues generated by the moving spot and that their spatiotemporal characteristics suggest detection by magnocellular processes. PMID- 10795632 TI - Technique to investigate the temporal phase shift between L- and M-cone inputs to the luminance mechanism AB - We describe a technique to estimate the intrinsic phase shift between long wavelength-cone (L-cone) and middle-wavelength-cone (M-cone) signals in the luminance mechanism with minimal contamination by chromatic mechanism(s). The technique can also estimate, simultaneously with the phase shift, the weight ratio of L and M cones for the luminance mechanism. We measured motion identification thresholds for a 1.0 cycle/deg, 12.0-Hz sinusoidal grating representing different vector directions in L- and M-cone contrast space. The physical phase of the L- and M-cone signals was varied over a broad range between -150 deg and +150 deg to investigate the effect on the threshold contours. The slope of the threshold contour in cone contrast space varied as a function of the physical phase. Estimates of the intrinsic phase shift between L and M cones are based on the change in slope of the threshold contour. The estimates are consistent with previous reports and show that whereas the L-cone signal lags behind the M-cone signal by approximately 35 deg for an orange background, the M cone signal lags behind the L-cone signal by approximately 8 deg for a green background. PMID- 10795633 TI - Numerical study of the displacement of a three-dimensional Gaussian beam transmitted at total internal reflection. Near-field applications AB - Longitudinal and transverse shifts of a light beam at total internal reflection was experimentally studied by far-field measurements on the reflected field. We propose to use a scanning tunneling optical microscope (STOM) to study these shifts in transmission, and we present a theoretical model of this proposed experiment to obtain a numerical estimation of these shifts. We study the reflection and the transmission of a three-dimensional polarized incident beam. We verify the validity of our formalism by studying the Goos-Hanchen shift in reflection and by comparing our results with published ones. Then we calculate STOM images of the transmitted field distribution. On the images the well-known Goos-Hanchen shift is easily observed. But we also encounter a smaller shift, perpendicular to the plane of incidence. This transverse shift was also observed in reflection by Imbert and Levy [Nouv. Rev. Opt. 6, 285 (1975)]. We study the variations of the two shifts versus various parameters such as the angle of incidence, the optical index, and the incident polarization. Then we discuss the feasibility of the near-field observation of these shifts. PMID- 10795634 TI - Quality parameters analysis of optical imaging systems with enhanced focal depth using the Wigner distribution function AB - An analysis of the Strehl ratio and the optical transfer function as imaging quality parameters of optical elements with enhanced focal length is carried out by employing the Wigner distribution function. To this end, we use four different pupil functions: a full circular aperture, a hyper-Gaussian aperture, a quartic phase plate, and a logarithmic phase mask. A comparison is performed between the quality parameters and test images formed by these pupil functions at different defocus distances. PMID- 10795635 TI - Second-order iterative approach to inverse scattering: numerical results AB - We introduce an iterative algorithm for the reconstruction of dielectric profile functions from scattered field data, in which each step corresponds to the solution of a quadratic inversion problem. This means that, at each iteration, we perform a second-order approximation of the scattering operator connecting the unknown profile to the data about a reference profile function. This procedure is then compared with a linear iterative inversion algorithm, and it is pointed out that, within a prescribed class of profile functions, the linear iterative inversion does not converge to the actual solution, whereas the proposed approach does. This feature can be explained by reference not only to the improved approximation provided by the addition of a further term for profile functions of a larger norm but also to the different classes of functions that can be reconstructed by either the linear or the quadratic model. Numerical examples of profile reconstruction in the scalar two-dimensional geometry, with far-zone scattered field data at a fixed frequency, confirm the theoretical analysis. PMID- 10795636 TI - Symmetries of cross-polarization diffraction coefficients of gratings AB - In a recent paper [J. Opt. Soc. Am. A 16, 1108 (1999)] Logofatu et al. demonstrated by experimental and numerical evidence that the 0th-order cross polarization (s to p and p to s) reflection coefficients of isotropic, symmetrical, surface-relief gratings in conical mount are identical. Here an analytical proof is given to show that the observed identity is merely a manifestation of the electromagnetic reciprocity theorem for the 0th-order diffraction of symmetrical gratings. The above result is further generalized to bianisotropic gratings, to the 0th-order cross-polarization transmission coefficients, and to the mth-order reflection and transmission coefficients when the wave vector of the incident plane wave and the negative of the wave vector of the mth reflected order are symmetrical with respect to the plane perpendicular to the grating grooves. PMID- 10795637 TI - Full angular profile of the coherent polarization opposition effect AB - We use the rigorous vector theory of weak photon localization for a semi-infinite medium composed of nonabsorbing Rayleigh scatterers to compute the full angular profile of the polarization opposition effect. The effect is caused by coherent backscattering of unpolarized incident light and accompanies the well-known backscattering intensity peak. PMID- 10795638 TI - Electromagnetic degrees of freedom of an optical system AB - We present a rigorous electromagnetic formalism for defining, evaluating, and optimizing the degrees of freedom of an optical system. The analysis is valid for the delivery of information with electromagnetic waves under arbitrary boundary conditions communicating between domains in three-dimensional space. We show that, although in principle there is an infinity of degrees of freedom, the effective number is finite owing to the presence of noise. This is in agreement with the restricted classical theories that showed this property for specific optical systems and within the scalar and paraxial approximations. We further show that the best transmitting and receiving functions are the solutions of well defined eigenvalue equations. The present approach is useful for understanding and designing modern optical systems for which the previous approaches are not applicable, as well as for application in inverse and synthesis problems. PMID- 10795639 TI - Generalized Fried parameter after adaptive optics partial wave-front compensation AB - Atmospheric turbulence imposes the resolution limit attainable by large ground based telescopes. This limit is lambda/r(0), where r(0) is the Fried parameter or seeing cell size. Working in the visible, adaptive optics systems can partially compensate for turbulence-induced distortions. By analogy with the Fried parameter, r(0), we have introduced a generalized Fried parameter, rho(0), that plays the same role as r(0) but in partial compensation. Using this parameter and the residual phase variance, we have described the phase structure function, estimated the point-spread function halo size, and derived an expression for the Strehl ratio as a function of the degree of compensation. Finally, it is shown that rho(0) represents the diameter of the coherent cells in the pupil domain. PMID- 10795640 TI - Wave-front sensing and deformable-mirror control in strong scintillation AB - Recent studies of coherent wave propagation through turbulence have shown that under conditions where scintillation is significant a continuous phase function does not in general exist, owing to the presence of branch points in the complex optical field. Because of branch points and the associated branch cuts, least squares approaches to wave-front reconstruction and deformable-mirror control can have large errors. Branch-point reconstructors are known to provide superior performance to least-squares reconstructors, but they require that branch points be explicitly detected. Detecting branch points is a significant practical impediment owing to spatial sampling and measurement noise in real wave-front sensors. Branch points are associated with real zeros in an optical field, and hence information about the phase of the field is encoded in the amplitude of the wave. We present a new wave-front-sensor processing algorithm that exploits this observation in the wave-front-reconstruction and deformable-mirror-control process. This algorithm jointly processes three intensity measurements by using light from the beacon field to develop a set of deformable-mirror actuator commands that are maximally consistent with three intensity measurements: (1) the entire wave-front-sensor image, (2) a pupil intensity image, and (3) a conventional image. Owing to the nonlinear nature of the resulting algorithm, we have used a simulation to evaluate performance. We find that in a focused laser beam projection paradigm that uses a point-source beacon, the new algorithm provides significantly improved performance over that of conventional Hartmann sensor least-squares deformable-mirror control based on centroid processing of wave-front-sensor outputs. The performance of the new algorithm approaches, the performance of an idealized branch-point reconstructor that requires pointwise phase differences for operation. PMID- 10795641 TI - Jones-matrix analysis with Pauli matrices: application to ellipsometry AB - The coherency matrix formalism based on Pauli matrices is applied to analyze a general ellipsometer that is described by Jones matrices. Here the Jones matrices are represented as sums of appropriate coefficients times the Pauli matrices and the identity matrix, and intensities are represented as traces of coherency matrices. This approach allows us not only to treat partial polarizations explicitly but also to take advantage of various identities to reduce to algebra the operations necessary for system analysis. A general expression is obtained for the intensity transmitted through a polarizer-sample-compensator-analyzer (PSCA) ellipsometer. This general expression is applied to an ideal PSCA ellipsometer, and then the results are reduced to describe several simpler but commonly used configurations. The results provide insight regarding general capabilities and limitations and allow us to compare different ellipsometer systems directly. Finally, this expression is extended to include artifacts, the explicit representation of which allows a complete determination of their defects. PMID- 10795642 TI - Limitations of interferometry due to the flicker noise of laser diodes AB - Interferometry with laser diodes is a cost-effective way to perform displacement measurement. The tunability of laser diodes is also of great interest in multiple wavelength interferometry. However, the additional flicker noise in the frequency noise spectrum of semiconductor lasers may become a limiting factor. Investigations on the limitations due to the 1/f noise of laser diodes are presented for both classical and multiple-wavelength interferometry. Measurements at the limit of the coherence length of laser diodes with the corresponding phase fluctuations are reported. The theoretical results are verified experimentally. PMID- 10795643 TI - Molecular fluorescence in the vicinity of a gradient-index medium AB - The problem of molecular fluorescence in the vicinity of a gradient-index medium is studied theoretically through classical modeling of a radiating dipole. A previously developed formulation involving the Green dyadic for an inhomogeneous medium is applied to the present problem. Normalized lifetimes for the admolecules are calculated and compared with those for a homogeneous medium. The results are illustrated by numerical examples assuming certain simple forms for the index profile. PMID- 10795644 TI - Seasonal allergic rhinitis and cognitive function. PMID- 10795645 TI - Regulation of B lymphocyte differentiation. AB - LEARNING OBJECTIVES: Type I hypersensitivity reactions uniquely involve the IgE class of immunoglobulins (Ig). IgE differs from other classes of Ig in that the majority of the antibodies are bound to high affinity IgE Fc(epsilon)Rs that are expressed on a variety of cell types. Some of these cell types, most notably, mast cells and basophils, are triggered to undergo rapid activation, degranulation, and release of bioactive mediators following binding of antigen to Fc(epsilon)RI-bound IgE. Because of the central role that IgE antibodies and these mediators play in the tissue injury typical of type I hypersensitivity, this article will review the various stages of B lymphocyte development, activation, and differentiation and comment, where appropriate on potential sites of deregulation in allergic disease. DATA SOURCES: A literature search of the stages of B lymphocyte differentiation with emphasis on events that concern IgE expression was performed. RESULTS: B lymphocyte differentiation into IgE expressing cells is dependent upon three types of signals. The first signal is delivered through the B cell antigen receptor and is pivotal in determining the antigenic specificity of the response. The second signal is provided primarily by cytokines derived from T helper 2 (TH2) cells, ie, interleukin (IL)-4 and IL-13. These cytokines are under tight regulation and their role appears to be the stimulation of transcription through the Ig constant region genes. Finally, the third signal is provided via the interaction between the constitutively expressed CD40 molecule on B lymphocytes and CD154 (CD40 ligand), a molecule expressed on T lymphocytes following activation. Elevated levels of IgE in atopic individuals may result from the preferential activation of TH2 cells. CONCLUSIONS: A greater understanding of the regulation of IgE expression may be central to the development of more effective immunotherapy strategies designed to attenuate IgE synthesis. PMID- 10795646 TI - Chronic oral mucosal ulceration in a 54-year-old female. AB - Pemphigus vulgaris most often begins in the mouth but is often overlooked in the differential diagnosis of chronic, multiple oral ulcerations and erosions. Accurate diagnosis requires perilesional biopsy including intact epithelium, submitted for hematoxylin and eosin as well as direct immunofluorescence staining. Early and aggressive treatment with moderate to high dose prednisone in combination with steroid sparing drugs such as azathioprine and mycophenolate allow complete remission in most patients. PMID- 10795647 TI - Cost-effectiveness analysis of budesonide aqueous nasal spray and fluticasone propionate nasal spray in the treatment of perennial allergic rhinitis. AB - BACKGROUND: An economic evaluation was performed analyzing direct medical costs in Canada for the treatment of perennial allergic rhinitis (PAR) with budesonide aqueous nasal spray and fluticasone propionate nasal spray. Three hundred fourteen patients with at least a 1-year history of PAR were randomized into a double-blind, parallel-group study of 6 weeks' duration. The treatments were daily doses of budesonide 256 microg, fluticasone propionate 200 microg, or placebo. Both active treatments produced significantly lower mean scores for overall nasal symptoms compared with placebo, and both were well tolerated. Budesonide was significantly more effective than fluticasone in reducing "blocked nose." METHOD: A retrospective cost-effectiveness analysis utilizing the clinical trial data was performed on the total costs of (1) budesonide-based and (2) fluticasone-based treatment strategies, including the relative importance of the drug costs in both strategies. RESULTS: The average treatment cost per patient in Canada over 12 months in the budesonide group was CAD 389.85 which was 23.3% lower than in the fluticasone group, which was CAD 508.06, due to lower drug acquisition costs (for the year 1998). CONCLUSION: Budesonide aqueous nasal spray was shown to be more cost-effective than fluticasone propionate nasal spray in the treatment of perennial allergic rhinitis. This result is valid in the province of Ontario, Canada and in many other settings with the same structure of relative prices. The result is mainly driven by a difference in drug cost. PMID- 10795648 TI - Effects of seasonal allergic rhinitis on selected cognitive abilities. AB - BACKGROUND: Many allergy patients complain of slowed thinking, memory problems, and difficulty sustaining attention during their allergy seasons. OBJECTIVE: This study evaluated the effect of symptomatic allergic rhinitis on speed of cognitive processing, ability to divide and sustain attention, working memory, and recent verbal memory. METHODS: Symptomatic ragweed-allergic rhinitis patients and nonatopic control subjects did cognitive testing in, out of, and in ragweed seasons. RESULTS: Test results indicate that, during ragweed seasons, allergic patients experience subtle slowed speed of cognitive processing but not deficits in attention and recent memory. Some patients also have difficulties in working memory. CONCLUSIONS: These findings suggest that having allergic reactions to ragweed pollen causes significant cognitive difficulties in a subgroup of patients. PMID- 10795649 TI - Eosinophil apoptosis in induced sputum from patients with seasonal allergic rhinitis and with asymptomatic and symptomatic asthma. AB - BACKGROUND: Eosinophilic inflammation is known to play an important role in the pathogenesis of allergic diseases. Apoptosis, a form of programmed cell death, is characterized by morphologic cell changes and leads to recognition and ingestion by macrophages. Apoptosis could be an important mechanism controlling the resolution of tissue eosinophilia. OBJECTIVE: This study was designed to investigate the presence of apoptotic eosinophils in induced sputum of patients with seasonal allergic rhinitis (SAR), when examined during natural pollen exposure and of patients with perennial asthma of different degrees of severity. METHODS: We recruited 11 patients with SAR to grass pollens, 26 patients with asymptomatic asthma (AA), and 18 patients with symptomatic asthma (SA). The severity of asthma was assessed by clinical scoring. Sputum was induced following a standard method and differential cell count was estimated. Eosinophils showing cell shrinkage and nuclear coalescence were classified as apoptotic. The number of apoptotic eosinophils was expressed as the percentage of total cells in sputum and as the proportion of apoptotic eosinophils relative to normal bilobed eosinophils ("apoptotic ratio"). RESULTS: We found the number of eosinophils in the SA group was significantly greater than that in the SAR and the AA groups (P < .001 and P < .0001 respectively). The number of apoptotic eosinophils in the AA group was significantly lower than that in the SAR group (P < .001) and in the SA group (P < .0001). The apoptotic ratio for eosinophils in the SAR group was significantly greater than in the AA group (P < .05) and in the SA group (P < .05). There was no difference in the apoptotic ratio between the AA and SA groups. CONCLUSIONS: This study confirms that apoptotic eosinophils are detectable in induced sputum of allergic patients. Further, the results of our study suggest that apoptosis could be an important mechanism in the control of acute eosinophilic inflammation in patients with SAR exposed to the sensitizing antigens. It appears that the apoptotic mechanism could be less effective in controlling tissue eosinophilia in asthmatic patients with chronic eosinophilic inflammation. PMID- 10795650 TI - Once-daily mometasone furoate dry powder inhaler in the treatment of patients with persistent asthma. AB - BACKGROUND: Although inhaled glucocorticoids are recommended for all stages of persistent asthma, compliance with long-term therapy is often poor, leading to significant morbidity and mortality. A simplified, once-daily dosing regimen may foster improved compliance. OBJECTIVE: To compare the efficacy and safety of once daily (AM) administration of mometasone furoate dry powder inhaler (MF DPI) 200 microg and 400 microg with placebo in patients with asthma previously maintained only on short-acting inhaled beta-adrenergic receptor agonists. METHODS: This was a 12-week, double-blind, placebo-controlled, parallel group study. The mean change from baseline to endpoint (last treatment visit) for FEV1 was the primary efficacy variable. RESULTS: At endpoint, both doses of MF DPI were significantly more effective than placebo (P < or = .05) in improving FEV1. Based on morning peak expiratory flow rate, once-daily MF DPI 400 microg was more effective than placebo (P < or = .001) at endpoint. Both active treatments also demonstrated improvement at endpoint in asthma symptom scores, physician-evaluated response to therapy and use of rescue medication. Although both MF DPI dosages were efficacious, MF DPI 400 microg provided additional improvement in some measures of pulmonary function (eg, morning PEFR) when these agents were administered once daily in the morning. Both doses of MF DPI were well tolerated and treatment related adverse events occurred at a similar incidence among the three treatment groups. CONCLUSIONS: The results of this study indicate that once-daily (AM) MF DPI provides a convenient and effective treatment option for patients with mild or moderate persistent asthma. PMID- 10795651 TI - In childhood asthma the degree of allergen-induced T-lymphocyte proliferation is related to serum IgE levels and to blood eosinophilia. AB - OBJECTIVE: To investigate whether the state of activation of circulating T-cells in childhood asthma could be related to serum IgE levels and/or to blood eosinophilia. METHODS: Seventeen atopic asthmatic children, sensitized to Dermatophagoides pteronyssinus (Der p), in stable condition at the time of the study and 15 sex-matched and age-matched controls were studied. The expression of activation surface markers (HLA-DR and CD25) on peripheral blood mononuclear cells (PBMCs) was tested by monoclonal antibodies and FACS analysis, while the PBMC proliferative response to Der p antigens was measured by tritiated thymidine (3HTdR) incorporation. RESULTS: As compared to controls, atopic children showed higher eosinophil counts (P < .01), similar lymphocyte counts (P > .1, each comparison) but higher proportion of HLA-DR+ and CD25+ T-lymphocytes (P < .05, each comparison). A significant Der p allergen-induced PBMC proliferation was observed in atopic children (P < .01) but not in controls (P > .1). Both in controls and in atopic children, no correlations were found between lymphocyte counts and eosinophil counts or total or allergen-specific IgE levels (P > .1, each comparison). In contrast, weak correlations were detected between the degree of allergen-induced PBMC proliferation and: a) allergen-specific IgE levels in serum (P < .05) and b) eosinophil counts (P < .05). CONCLUSION: These data support the concept that the degree of activation of allergen-specific T lymphocytes in blood may reflect the intensity of allergic sensitization in childhood asthma. PMID- 10795652 TI - Sensitization to aeroallergens among patients with allergic rhinitis in a desert environment. AB - BACKGROUND: The causative allergens of allergic rhinitis in desert environments are uncertain. OBJECTIVE: To determine the sensitizing aeroallergens in patients with allergic rhinitis in Kuwait, a desert country. METHODS: A total of 706 patients aged 6 to 64 years (mean 34.3 years) with allergic rhinitis were studied. Sera from the patients were screened for specific IgE to 14 inhalant allergens by the CAP-RAST method. RESULTS: Specific IgE to any allergen was detected in 86.3% of patients. The prevalence rates for allergen groups were: pollens (77.3%), house dust (62.3%), and molds (14.7%). The individual allergens with the highest positive rates were pollens of the weed Chenopodium (64.3%); Bermuda grass (55.0%), and Prosopis tree (50.3%). These plants were all imported and cultivated for the purpose of "greening" the desert. German cockroach (48.2%) and house dust mites (32.4% to 39.2%) were the most prevalent indoor sensitizers. With the exception of the molds, sensitization rates were higher for males than females. The youngest age group (6 to 17 years) had significantly higher sensitization rates than the older ones, particularly with respect to the molds (P < .01 to .001). Severe sensitization was more common with Alternaria than the other allergens and in general mold sensitization was more frequently associated with severe symptoms. Polysensitization was very common, with 81.8% of all sensitized patients positive to more than one allergen. CONCLUSIONS: Pollens of the local horticultural plants are the main sensitizing allergens among patients with allergic rhinitis in this desert environment. The practices that "green" the desert seem to also encourage allergen sensitization. PMID- 10795653 TI - Incidence of sensitivity to Anisakis simplex in a risk population of fishermen/fishmongers. AB - BACKGROUND: Anisakis simplex, a fish and cephalopodes parasite, can cause either gastrointestinal symptoms or allergic reactions in humans on eating/handling contaminated fish. OBJECTIVE: The aim of our study was to determine the capacity of Anisakis simplex to induce specific IgE production and allergic reactions following eating and handling fish in a population at risk. METHODS: We determined the levels of total IgE, specific IgE, and eosinophil count in 28 fishermen/fishmongers (group A) and 15 healthy donors (group B). A skin prick test (SPT) with extracts from Anisakis and the most common species of fish in our country, were also carried out. RESULTS: Specific IgE to Anisakis were found in 14 subjects of group A (13 of them had a positive SPT to the same extract) and none of group B (only one subject had a positive SPT). The SPT with fish extracts was positive in 4 patients of group A but in none of group B. Subjects in group A with specific IgE to Anisakis showed higher total IgE levels and eosinophil counts compared with either other individuals of the same group or to those of group B. CONCLUSIONS: These results indicate that fishermen/fishmongers are a population at risk for Anisakis simplex sensitization and suggest that this kind of sensitization should also be investigated not only in subjects like fishermen/fishmongers who live in countries where fish is likely to be contaminated with Anisakis simplex parasites, but also in those who handle fish for other reasons. PMID- 10795654 TI - Gender differences in response to the multitest CMI skin test in the general population. AB - BACKGROUND: Previous studies of gender differences in response to the Multitest CMI skin test have produced conflicting results. OBJECTIVE: To determine whether gender is associated with response to the Multitest CMI skin test. METHODS: Two hundred ninety-seven adults, aged 18 to 64 years, recruited originally for a study of the immune effects associated with living near a hazardous waste site containing primarily organochlorine pesticides, underwent a skin test using the Multitest CMI skin test. Six of seven antigens were tested: tetanus toxoid, diphtheria toxoid, Candida, Tricophyton, Streptococcus, and Proteus. The tuberculin antigen was excluded. Lymphocyte function was also evaluated in vitro using standardized methods of mitogen stimulation with phytohemaglutinin (PHA), concanavalin A (CON-A), and pokeweed mitogen. RESULTS: The frequency of positive responses to the skin tests was significantly (P < .001) higher among males (80.4%) than among females (55.7%). Males were more likely than females to respond to all six antigens tested (P < .05). The mean diameter of positive skin test measurements for males statistically significantly (P < .05) exceeded female responses for tetanus and diphtheria. Although not statistically significant, male response size exceeded that of females for all other antigens except Trycophyton. Controlling for age, race, smoking, income, and plasma DDE levels did not change these results. Skin test positivity was not associated with mitogen stimulation assay results overall or within gender groups. CONCLUSION: Significant gender differences in response to the Multitest CMI skin test could limit its use as a marker of anergy in general population studies. PMID- 10795656 TI - Fennel, cucumber, and melon allergy successfully treated with pollen-specific injection immunotherapy. AB - BACKGROUND: In subjects with both pollinosis and vegetable food allergy, most allergenic epitopes of fruits and vegetables are present in pollen. A recent study showed a marked reduction or a total disappearance of apple-induced oral allergy syndrome in patients receiving injection immunotherapy with birch pollen extracts. OBJECTIVE: To assess whether vegetable food allergy following other kinds of primary pollinosis may be successfully treated with pollen-specific immunotherapy. METHODS: A 34-year-old woman with long-standing pollinosis and typical oral allergy syndrome (OAS) with the ingestion of both fennel and cucumber and whose OAS was associated with immediate laryngeal edema after the ingestion of melon, was treated with two commercial depot aluminum hydroxide adsorbed extracts of 1 grass pollen and 2 mugwort pollen 50% + ragweed pollen 50%. RESULTS: After 36 months of injection specific immunotherapy, the patient was able to tolerate both fresh fennel and cucumber without consequence on open oral challenge tests. After 43 months of immunotherapy, the patient tolerated fresh melon as well on open oral challenge. She has re-introduced these vegetables in her normal diet. Skin tests showed no reactivity to fresh fennel and there was a reduction of the wheal induced by fresh cucumber. CONCLUSION: Vegetable food allergy following primary sensitization to pollens, other than birch, may also be effectively reduced by pollen-specific injection immunotherapy. PMID- 10795655 TI - Onset-of-action for antihistamine and decongestant combinations during an outdoor challenge. AB - BACKGROUND: Medications containing a combination antihistamine-decongestant are commonly used for allergic rhinitis yet onset-of-action comparisons for symptom relief after a single dose have not been performed. OBJECTIVE: To determine the onset of symptom relief and efficacy of antihistamine-decongestant medications (acrivastine-pseudoephedrine and loratadine-pseudoephedrine) compared with placebo in an outdoor park. METHODS: This study was conducted during the spring of 1997 using a double-blind, placebo-controlled design. Patients completed baseline rhinitis symptom diaries from 7:30 to 9:30 AM. Subjects with qualifying symptom scores received one dose of either acrivastine-pseudoephedrine, loratadine-pseudoephedrine, or placebo at 10:00 AM. Symptom diaries were recorded for the next 4 hours. RESULTS: Of 593 patients randomized to treatment, 592 were included in efficacy analysis. Acrivastine-pseudoephedrine and loratadine pseudoephedrine demonstrated a mean onset-of-action by 45 and 30 minutes respectively for total symptom and rhinitis symptom scores for the five sites. Onset-of-action for nasal congestion scores was 45 minutes for both medications. Sites having higher pollen exposure (>100 pollen grains over 6 hours) demonstrated a difference between the antihistamine combinations: acrivastine pseudoephedrine had an onset of action at 45 minutes for total symptom and rhinitis symptom scores, and 15 minutes for nasal congestion scores whereas loratadine-pseudoephedrine had onset-of-action for nasal congestion score of 105 minutes but failed to reach significance at any timepoint for total symptom and rhinitis symptom scores. CONCLUSIONS: Both antihistamine-decongestant combinations demonstrate an onset-of-action within 60 minutes of administration but under conditions of higher pollen exposure, the acrivastine combination was more effective for total symptoms, rhinitis symptoms, and nasal congestion with an onset-of-action within 45 minutes for rhinitis symptoms and 15 minutes for congestion. PMID- 10795657 TI - Causes of low serum albumin in peritoneal dialysis patients. PMID- 10795658 TI - Tuberculous peritonitis in patients on continuous ambulatory peritoneal dialysis. PMID- 10795659 TI - Helicobacter pylori infection in patients with end-stage renal disease. PMID- 10795660 TI - Removal of Foscarnet by hemodialysis using dialysate-side values. AB - Foscarnet is an antiviral agent widely used in the treatment of cytomegalovirus (CMV) infection. We describe a cardiac transplant patient, who while being maintained with hemodialysis because of tobramycin-induced acute renal failure, was given Foscarnet for disseminated CMV infection. Using dialysate-side clearance methodology, we found the dialyzer clearance of Foscarnet to be in the order of 89 ml/min. PMID- 10795661 TI - The impact of equations on calculation of lean body mass by bioelectrical impedance analysis in RDT patients. AB - Several equations are available to derive lean body mass (LBM) from bioelectrical impedance analysis (BIA). The purpose of this study was to investigate in dialysis patients the impact of the equation used on the outcome of LBM assessment. To avoid dyshydration as a confounder, vena cava diameter measurement was used to assess normohydration in the 21 patients studied. Five equations were compared. In a previously published study to assess total body water using antipyrine as a gold standard, Deurenberg's formula was advocated to be used in the estimation of LBM by BIA. Therefore, this formula was used as a basis for comparison with the other four equations. One equation gave results comparable to those obtained by Deurenberg's formula. Despite high correlations and agreement according to Bland and Altman analysis, the other three equations showed a significant difference with Deurenberg-derived LBM. Thus, the equation used has a major impact on the outcome of LBM estimations. PMID- 10795662 TI - The effect of hemodialysis and acetate-free biofiltration on anemia. AB - The authors monitored, for a period of 12 months, anemia-, nutrition-, and free radical-related parameters and the rHuEPO dose required to maintain target hemoglobin (Hb) in 20 patients with chronic renal failure. Ten patients each were randomized for treatment by either acetate-free biofiltration (AFB) or low-flux hemodialysis (HD). At baseline, Hb levels were 102+/-2 (AFB) vs. 98+/-2 g/L (HD) (not significant difference, NS), the rHuEPO dose was 4050+/-976 vs. 5100+/-1538 lU/week (NS). Compared with baseline and with HD, lower rHuEPO doses were required during AFB at months 8, 9, 10 and 11, and 12 when they were 2100+/-510 (AFB) vs. 6000+/-1153 (HD), p=0.008. Prealbumin, transferrin and cholinesterase levels rose in the AFB group. Kt/V, albumin, transferrin saturation, aluminium, bicarbonate in serum, superoxide dismutase and glutathione peroxidase in erythrocytes, and malondialdehyde and antioxidant capacity in plasma did not differ between the AFB and HD groups. In terms of anemia control, AFB using an AN69 membrane was found to be more advantageous than low-flux HD, AFB improves some nutritional parameters. The compared methods do not differ in their effect on lipid peroxidation and the antioxidant system. PMID- 10795663 TI - Prevention of nosocomial transmission of hepatitis C infection in a hemodialysis unit. A prospective study. AB - Hepatitis C virus (HCV) infection in hemodialysis patients can be transmitted by transfusions and nosocomially. A high prevalence of HCV infection, over 50%, was demonstrated in our hemodialysis (HD) unit. In order to prevent the nosocomial spread of HCV infection in the HD unit a prospective study was begun separating anti-HCV positive patients from the negative ones. A total of 170 patients (83 anti-HCV positive) started this study in September 1994 and were followed for 4 years. A separate room and dedicated equipment were assigned to anti-HCV positive and anti-HCV negative patients. Of those 170 patients there were 15 hepatitis B virus (HBV) positive patients, 5 of whom were anti-HCV positive, who were treated in a separate room on dialysis equipment for anti-HCV positive or negative patients. Application of general precautions, as recommended by the Center for Disease Control and Prevention (CDC), was reinforced. During the first 12 weeks after implementing the precautions seven more anti-HCV positive patients were detected, and by December 1995 another two HCV infected patients were found. The follow-up included all changes in HD population treated until the end of 1998. The incidence of seroconversion to HCV was 12.9% in 1995, 7.1% in 1996, 5.0% in 1997, and 6.6% in 1998. The higher incidence of seroconversion in September to November 1994 was probably due to the nosocomial infection being in the incubation period at the time of isolation. This prospective study in a large HD unit with a high prevalence of HCV infection demonstrates a relatively successful prevention of HCV spread. Procedure-related transmission of HCV in hemodialysis could be prevented by rigorous application of universal precautions as recommended by the CDC. As a second line of prevention, in highly burdened dialysis centers, segregation of HCV positive patients can help control nosocomial transmission. PMID- 10795664 TI - Extracorporeal circulation in ewe's foetus: towards a reliable foetal cardiac surgery protocol. A comparison of two cases. AB - Foetal cardiac surgery is the ultimate goal in the treatment of congenital cardiac malformations. The aim of our research is to elucidate some of the features of the necessarily invasive experimental protocol to be used in an animal model of foetal cardiac surgery. In particular, we assessed the foetal placental reactivity to prolonged cardiac bypass in steady-flow conditions. METHODS: Two cases were selected to show the outcome of prolonged (> 30 minutes) extracorporeal circulation (ECC) instituted without oxygenator under steady-flow assistance. Following the instrumentation of the animal (placement of pressure, flow and myocardial fiber length transducers) and the baseline recordings, a 60 minute bypass period was established with an axial turbopump (Hemopump 14 Fr), after systemic heparinisation and artero-venous cannulation. At the end of the circulatory assistance, the cannulae were removed and a 90 minute observation period followed. The cardiac function was assessed by means of indirectly obtained P-V loops. RESULTS: Case A showed a marked reduction in the end-systolic pressure-volume relationship (ESPVR) during ECC, corresponding to a rightward shift of the P-V loop, with a gradual recovery after the assisted circulation. On the contrary, case B was subjected to progressive placental dysfunction, as evidenced by haemogasanalytical data. Consequently, the haemodynamic data also outlined a negative outcome, with high ESPVR values after bypass. CONCLUSIONS: The present study, while confirming the possibility of cardiac intervention in the foetus, underlines the critical role of minimally invasive protocol to limit both foetal stress and placental dysfunction. PMID- 10795665 TI - Intraindividual comparison of the impact of two selective apheresis methods (DALI and HELP) on the coagulation system. AB - We performed an intraindividual comparison of the effect on the coagulation system of two selective apheresis procedures: Direct Adsorption of Lipoproteins (DALI) and Heparin-induced Lipoprotein Fibrinogen Precipitation (HELP). Six patients suffering from heterozygous familial hypercholesterolemia have been treated with 2 sessions of each procedure. Anticoagulation was carried out according to usual recommendations. Blood samples were taken before, immediately after and on the second day after the sessions. We assessed global coagulation tests (prothrombin time, activated partial thromboplastin time), fibrinogen, prothrombin fragment F 1 + 2 and a variety of factors (Factors II, V, VII, XIII, IX, X, XI, XII, XIIa; von Willebrand Factor; collagen-binding activity, prekallikrein, high-molecular weight kininogen) and antagonists (antithrombin III, protein S activity, free protein S). In fact, all parameters measured have been influenced by the apheresis treatment. Fibrinogen is lowered more by HELP which also has a more definite impact on factors belonging to the prothrombin complex (II, VII, X). In contrast, the major effects of the DALI system have been seen on the intrinsic pathway of the coagulation system (IX, XI, prekallikrein, high-molecular-weight kininogen). With both systems, no increases in activated Factor XII or in prothrombin fragment F 1 + 2 have been observed. These data provide a solid basis for individual adaptations of anticoagulant doses. PMID- 10795666 TI - Free nerve endings in the medial and posteromedial capsuloligamentous complexes: occurrence and distribution. AB - Free nerve endings (FNEs) of type IVa play a distinctive role in the articular nociceptive and sensorimotor system of the knee. This study qualitatively and quantitatively analyzed FNEs in the medial and posteromedial capsuloligamentous complexes. Biopsy specimens from ten precisely defined anatomical locations were taken from seven fresh cadaver knee joints. The specimens were fixed with 4% formaldehyde solution and stained with hematoxylin-eosin. The results were examined using immunohistochemistry. The occurrence of FNEs is described in combination with their specific pattern of distribution. A high number of FNEs were found in all investigated elements with a maximum relative density in the insertion of the semimembranosus muscle in the direct attachment on the tibial margin. The number was lowest in the superficial medial collateral ligament. The results were correlated with anatomical and biomechanical functions of the stabilizing effect of the medial capsuloligamentous complex. Our findings indicate that lesions and surgical procedures can alter normal sensory feedback and coordination by modifying the use of muscle fiber during specific movements. PMID- 10795667 TI - Comparison of eccentric and concentric screw placement for hamstring graft fixation in the tibial tunnel. AB - Interference screw fixation of four-strand hamstring grafts for ACL reconstruction has recently been introduced. By this method, the interference screw is placed in the tibial and femoral tunnels eccentric (adjacent) to the bundled limbs of the graft. In order to maximize the graft to tunnel contact to promote biological fixation, it is proposed to place the screw concentrically in the tunnel, in the middle of the four limbs of the graft, pressing each limb of the graft into the tunnel wall. This would be difficult to do in the proximal, folded end of the four limb graft situated in the femoral tunnel but can be done easily in the tibial tunnel. The purpose of this study was to evaluate the effect of screw placement on the stiffness, yield load, and ultimate load of hamstring graft fixation in the tibial tunnel. Five pairs of human knees were used for the study. Pull out tests were performed using an MTS system, pulling along the axis of the tibial tunnel. Tibial fixation stiffness was greater using concentric screw placement (P < 0.05) although there was no statistical difference in yield load, slippage, or ultimate load. PMID- 10795668 TI - A decision-making scheme for returning patients to high-level activity with nonoperative treatment after anterior cruciate ligament rupture. AB - This report describes the development and current use of decision-making criteria for returning patients to high-level physical activity with nonoperative management of anterior cruciate ligament ruptures, and presents the results of treatment for patients who met our criteria as candidates for nonoperative rehabilitation and attempted to return to high-level physical activity with nonoperative management. The screening examination consists of four one-legged hop tests, the incidence of knee giving-way, a self-report functional survey, and a self-report global knee function rating. We screened 93 consecutive patients with acute unilateral anterior cruciate ligament rupture, classifying them as either candidates (n = 39, 42%) or noncandidates (n = 54, 58%) for nonoperative management. Of the 39 rehabilitation candidates 28 chose nonoperative management and returned to preinjury activity levels, 22 of whom (79%) returned to preinjury activity levels without further episodes of instability or a reduction in functional status. No patient sustained additional articular or meniscal damage as a result of rehabilitation or return to activity. The decision-making scheme described in this study shows promise in determining who can safely postpone surgical reconstruction and temporarily return to physically demanding activities. Continued study to refine and further validate the decision-making scheme is recommended. PMID- 10795669 TI - Spontaneous osteonecrosis of the medial compartment of the knee: a MRI follow-up after conservative and operative treatment, preliminary results. AB - Symptoms in patients suffering from spontaneous osteonecrosis of the knee (SONK) may be reduced by high tibial osteotomy (HTO). However, the fate of the necrotic lesion is unknown and needs further investigation. We conducted a prospective study to evaluate magnetic resonance imaging (MRI) changes after operative and nonoperative treatment. Ten consecutive patients suffering from SONK of the medial compartment were given two treatment options: either HTO (n = 6) or conservative treatment with partial weight bearing for 3 months (n = 4). We measured the greatest extent of well-defined subchondral low signal intensity abnormality, considered to represent necrosis, and the surrounding area of intermediate signal intensity, considered to represent perifocal bone marrow edema, on T1-weighted coronal MRI images before and after treatment. The MRI follow-up period was 17.5 months (range 12-27) in the HTO group and 14.5 months (range 8-25) in the nonoperative group. At follow-up the MRI evaluation revealed a decrease in the low signal intense areas (necrosis) in five of the six patients in the HTO group. Only one of the four nonoperative patients showed a decrease in the low signal intense area. The intermediate intense areas (edema) decreased in all patients in the HTO group and in three of four in the nonoperative group. The mean decrease in the area of perifocal edema was significantly greater in the HTO group than in the nonoperative group (P = 0.019). No statistically significant difference was found for the area of necrosis between the two groups (P = 0.171). A clinical improvement was observed in all patients of the HTO group but in only two of the four patients of the nonoperative group. We conclude that the decrease in perifocal bone marrow edema seems to be associated with improved patient comfort. The MRI appearance of the necrotic lesion does not alter with either treatment mode. PMID- 10795670 TI - Driving reaction time after right knee arthroscopy. AB - Driving reaction times of 30 right knee arthroscopy patients were measured using a computer-linked car simulator. Each patient was tested pre-operatively and 1 week after and 4 weeks after arthroscopy. As controls, 25 normal subjects were also tested. In the control group the average reaction time was 634 ms; the measurements at 1 week and at 4 weeks were 550 ms and 582 ms, respectively. In the arthroscopy group the average reaction time pre-operatively was 736 ms; the measurements 1 week and 4 weeks post-operatively were 920 ms and 685 ms, respectively. Two clinical tests (the stepping and standing tests) were also performed at each assessment. Statistical analysis showed a good correlation between these and reaction time. We conclude that it is appropriate for patients to delay their return to driving for at least 1 week, and that the actual timing of return to driving may be determined by performance on these two clinical tests. PMID- 10795671 TI - Effects of sectioning the posterolateral structures on knee kinematics and in situ forces in the posterior cruciate ligament. AB - The objective of this study was to determine the effects of sectioning the posterolateral structures (PLS) on knee kinematics and in situ forces in the posterior cruciate ligament (PCL) in response to external and simulated muscle loads. Ten human cadaveric knees were tested using a robotic/universal force moment sensor testing system. The knees were subjected to three loading conditions: (a) 134-N posterior tibial load, (b) 5-Nm external tibial torque, and (c) isolated hamstring load (40 N biceps/40 N semimembranosus). The knee kinematics and in situ forces in the PCL for the intact and PLS-deficient knee conditions were determined at full extension, 30 degrees, 60 degrees, 90 degrees, and 120 degrees of knee flexion. Under posterior tibial loading posterior tibial translation with PLS deficiency increased significantly at all flexion angles by 5.5+/-1.5 mm to 0.8+/-1.2 mm at full extension and 90 degrees, respectively. The corresponding in situ forces in the PCL increased by 17-19 N at full extension and 30 degrees of knee flexion. Under the external tibial torque, external tibial rotation increased significantly with PLS deficiency by 15.1+/-1.6 degrees at 30 degrees of flexion to 7.7+/-3.5 degrees at 90 degrees, with the in situ forces in the PCL increasing by 15-90 N. The largest increase occurred at 60 degrees to 120 degrees of knee flexion, representing forces two to six times of those in the intact knee. Under the simulated hamstring load, posterior tibial translation and external tibial and varus rotations also increased significantly at all knee flexion angles with PLS deficiency, but this was not so for the in situ forces in the PCL. Our data suggest that injuries to the PLS put the PCL and other soft tissue structures at increased risk of injury due to increased knee motion and the elevated in situ forces in the PCL. PMID- 10795672 TI - Algorithm for establishing the indication for knee arthroscopy in children: a comparison of adolescent and preadolescent children. AB - The aim of this study was to select children with pathological lesions of the intra-articular structures from children with identical complaints but with no pathological intra-articular changes. The younger the child, the more difficult it is to make the diagnosis, and the expected distribution of pathology changes increasingly. This is particularly stressed in children aged younger than 13 years. Synovial inflammatory alterations are more frequent, and osteochondral and chondral fractures appear to be more problematic than meniscal and cruciate ligament lesions. Before establishing the indication for knee arthroscopy it is mandatory to implement the algorithm of diagnostic and conservative therapeutic procedures. The indication for knee arthroscopy is considered in cases when complaints persist after conservative treatment, a lesion of intra-articular segments is suspected, and the pathological condition is deemed arthroscopically treatable. Arthroscopy before conservative treatment is justified only in acute cases. PMID- 10795673 TI - Treatment of intrasubstance meniscal lesions: a randomized prospective study of four different methods. AB - This study examined the effect of four different methods for treating intrasubstance meniscal lesions. Forty patients (21 men, 19 women; age 30.4 years, range 16-50) with an isolated and symptomatic painful horizontal grade 2 meniscal lesion on the medial side (documented with MRI) were included. Patients were randomly assigned by the birth date to one of four treatment groups: group A, conservative therapy (n = 12); group B, arthroscopic suture repair with access channels (n = 10); group C, arthroscopic minimal central resection, intrameniscal fibrin clot and suture repair (n = 7); and group D, arthroscopic partial meniscectomy (n = 11). The average length of follow-up was 26.5 months (range 12 38 months). Follow-up evaluation consisted of clinical examination with the findings recorded according to the IKDC protocol, radiographs, and control MRI. Group A had 75% normal or nearly normal final evaluation at follow-up, group B 90%, group C 43%, and group D 100% normal or nearly normal at follow-up. These short-term results indicate that intrasubstance meniscal lesions can be treated best by performing partial meniscectomy. To preserve the important function of the meniscus, arthroscopic suture repair with access channels might give even better medium- to long-term results. Conservative treatment is often not satisfactory. Additionally, our findings show that MRI examinations are not superior to accurate clinical examinations. PMID- 10795674 TI - Isokinetic thigh muscle performance after long-term recovery from patellar dislocation. AB - Eighty-two patients (50 women, 32 men) underwent isokinetic muscle testing on average 13 years after a conservatively treated unilateral primary patellar dislocation. Three study groups were formed according to the natural history of recovery: group A (n = 32), patients with only primary conservative treatment; group B (n = 34) patients with conservative (group B1; n = 24) or surgical (group B2; n = 10) treatment of redislocations; group C (n = 16) patients with other residual complaints (anterior knee, pain subluxations) requiring surgery. The Cybex 6000 dynamometer system was used as the testing machine for quadriceps and hamstrings muscles, with proportional deficits of peak torque as the test parameter. Isokinetic testing revealed both quadriceps and hamstring muscle atrophy even after long-term recovery from injury. There were statistically significant differences between the three study groups at both tested speeds of quadriceps muscles (60 rad/s, P < 0.002; 180 rad/s, P < 0.009). Groups B1 and B2 presented similar results. The muscle performance findings are probably due to more than one factor: primary immobilization, poor outcome, patellofemoral degeneration, redislocations, and residual knee complaints followed by surgery and deficiency in motor control of thigh muscle had--together or separately--an effect on muscle performance. PMID- 10795675 TI - Rupture of the pectoralis major: a meta-analysis of 112 cases. AB - Of about 150 cases reported in the literature on pectoralis major ruptures, 108 were selected as presenting enough data to be analyzed for cause, rupture site, injury mechanism, and treatment outcome. We added data on four of our own cases reported here. All patients yet reported have been men. Rupture of the PM occurs most commonly in sports during weight training, weight lifting, or wrestling when the arm is externally rotated and abducted. Most reported ruptures are complete and are located at the insertion to the humerus. Work-related injuries occur more often at the musculo-tendinous junction. The prognosis is related neither to the age of the patient nor to the location of the rupture. Surgical treatment, preferably within the first 8 weeks after the injury, has a significantly better outcome than conservative treatment or delayed repair. PMID- 10795676 TI - Localization of growth factors in the reconstructed anterior cruciate ligament: immunohistological study in dogs. AB - This study was designed to examine localization of the growth factors in the autogenous patellar tendon graft used to reconstruct the anterior cruciate ligament (ACL) in the canine model. Among the various growth factors, basic fibroblast growth factor, transforming growth factor-beta1, and platelet-derived growth factor were selected for analysis as potential factors that regulate graft remodeling processes. In the control patellar tendon and the ACL only basic fibroblast growth factor was positively stained. In the reconstructed graft increased levels of staining for all the three factors were observed in the early postoperative period, reaching the greatest expression 3 weeks after implantation. Thereafter immunoreactivity of these growth factors decreased and returned to the preoperative levels, which were similar to that of the control ACL 12 weeks postoperatively. This rapid reduction in the level of their localization indicates that once the extrinsic cells are infiltrated to the graft and revascularization completed, these growth factors may have less significance for subsequent remodeling. PMID- 10795677 TI - Acute arthroscopic Bankart repair? PMID- 10795678 TI - Properties of the trihydroxytoluene oxygenase from Burkholderia cepacia R34: an extradiol dioxygenase from the 2,4-dinitrotoluene pathway. AB - Burkholderia cepacia R34 mineralizes 2,4-dinitrotoluene via an oxidative pathway. The initial steps in the degradative pathway lead to formation of 2,4,5 trihydroxytoluene, which serves as the substrate for the ring cleavage dioxygenase. The trihydroxylated substrate differs from the usual substituted catechols found in pathways for aromatic compound degradation. To determine whether the characteristics of the trihydroxytoluene oxygenase reflect the unusual ring cleavage substrate of the 2,4-dinitrotoluene pathway, the gene encoding trihydroxytoluene oxygenase (dntD) was cloned and sequenced, and ring cleavage activity determined from recombinant bacteria carrying the cloned gene. The findings were compared to the trihydroxytoluene oxygenase from Burkholderia sp. strain DNT and to other previously described ring cleavage dioxygenases. The comparison revealed that only 60% identity was shared by the two trihydroxytoluene oxygenases, but the amino acid residues involved in cofactor binding, catalysis, and protein folding were conserved in the DntD sequence. The enzyme catalyzed meta-fission of trihydroxytoluene as well as the substrate analogues 1,2,4-benzenetriol, catechol, 3-methylcatechol, 4-methylcatechol, 3 chlorocatechol, 4-chlorocatechol and 2,3-dihydroxybiphenyl. However, results from enzyme assays indicated a strong preference for trihydroxytoluene, implying that it was the native substrate for the enzyme. The apparent enzyme specificity, its similarity to the trihydroxytoluene oxygenase from Burkholderia sp. strain DNT, and the distant genetic relationship to other ring cleavage enzymes suggest that dntD evolved expressly to carry out trihydroxytoluene transformation. PMID- 10795679 TI - Two distinct pathways for anaerobic degradation of aromatic compounds in the denitrifying bacterium Thauera aromatica strain AR-1. AB - Denitrifying bacteria degrade many different aromatic compounds anaerobically via the well-described benzoyl-CoA pathway. We have shown recently that the denitrifiers Azoarcus anaerobius and Thauera aromatica strain AR-1 use a different pathway for anaerobic degradation of resorcinol (1,3-dihydroxybenzene) and 3,5-dihydroxybenzoate, respectively. Both substrates are converted to hydroxyhydroquinone (1,2,4-trihydroxybenzene). In the membrane fraction of T. aromatica strain AR-1 cells grown with 3,5-dihydroxybenzoate, a hydroxyhydroquinone-dehydrogenating activity of 74 nmol min(-1)(mg protein)-1 was found. This activity was significantly lower in benzoate-grown cells. Benzoate grown cells were not induced for degradation of 3,5-dihydroxybenzoate, and cells grown with 3,5-dihydroxybenzoate degraded benzoate only at a very low rate. With a substrate mixture of benzoate plus 3,5-dihydroxybenzoate, the cells showed diauxic growth. Benzoate was degraded first, while complete degradation of 3,5 dihydroxybenzoate occurred only after a long lag phase. The 3,5-dihydroxybenzoate oxidizing and the hydroxyhydroquinone-dehydrogenating activities were fully induced only during 3,5-dihydroxybenzoate degradation. Synthesis of benzoyl-CoA reductase appeared to be significantly lower in 3,5-dihydroxybenzoate-grown cells as shown by immunoblotting. These results confirm that T. aromatica strain AR-1 harbors, in addition to the benzoyl-CoA pathway, a second, mechanistically distinct pathway for anaerobic degradation of aromatic compounds. This pathway is inducible and subject to catabolite repression by benzoate. PMID- 10795680 TI - Cyanobacterial diversity in geographically related and distant host plants of the genus Gunnera. AB - The diversity among 45 cyanobacterial isolates from 11 different Gunnera species originating from different geographical areas was examined. By means of polymerase chain reaction (PCR) fingerprinting with short tandemly repeated repetitive (STRR) sequences as primers, ten groups of symbiotic cyanobacteria and five unique isolates not belonging to a particular group were identified. Most groups were restricted to one geographical area, indicating a limited distribution of related cyanobacterial strains. An extensive cyanobacterial diversity was found both within and between the 11 different Gunnera species. Within a particular plant and even within the same stem gland, more than one cyanobacterial strain at a time could be present. These results indicate a low specificity in Gunnera-Nostoc symbiosis. PMID- 10795681 TI - Purification and properties of the first-identified, archaeal, ATP-dependent 6 phosphofructokinase, an extremely thermophilic non-allosteric enzyme, from the hyperthermophile Desulfurococcus amylolyticus. AB - The ATP-dependent 6-phosphofructokinase (ATP-PFK) of the hyperthermophilic archaeon Desulfurococcus amylolyticus was purified 1,500-fold to homogeneity. The enzyme had an apparent molecular mass of 140 kDa and was composed of a single type of subunit of 33 kDa suggesting a homotetrameric (alpha4) structure. The N terminal amino acid sequence did not show significant similarity to ATP-PFKs isolated from eubacteria and eukarya. Kinetic constants of the enzyme were determined for both reaction directions at pH 6 and at 85 degrees C. Rate dependence on all substrates followed Michaelis-Menten kinetics. The apparent K(m)s for ATP and fructose 6-phosphate (forward reaction) were 0.28 and 1.17 mM, respectively; the apparent V(max) was about 41 U/mg. ATP could not be replaced by pyrophosphate (PPi) or ADP as phosphoryl donor, thus defining the enzyme as an ATP-dependent PFK. In addition to ATP (100%), the enzyme accepted GTP (97%), ITP (130%), UTP (84%), CTP (55%) and, less effectively, acetyl phosphate (13%) as phosphoryl donors. Enzyme activity was not allosterically regulated by classical effectors of ATP-PFKs such as ADP, AMP, and phosphoenolpyruvate or citrate. The enzyme also catalysed in vitro the reverse reaction with an apparent K(m) for fructose-1,6-bisphosphate and ADP of 16.7 and 0.5 mM, respectively, and an apparent V(max) of about 4.5 U/mg. Divalent cations were required for maximal activity; Mg2+, which was most effective, could be replaced partially by Ni2+, Mn2+ or Co2+. The enzyme had a temperature optimum of 90 degrees C and showed a significant thermostability up to 100 degrees C, which is in accordance with its physiological function under hyperthermophilic conditions. This is the first description of an ATP-dependent PFK from the domain of archaea, characterized as an extremely thermophilic, non-allosteric enzyme. PMID- 10795682 TI - Analysis of the cleavage site specificity of the endopeptidase involved in the maturation of the large subunit of hydrogenase 3 from Escherichia coli. AB - The maturation of [NiFe]-hydrogenases is a catalysed process in which the activities of at least seven proteins are involved. The last step consists of the endoproteolytic cleavage of the precursor of the large subunit after the [NiFe] metal centre has been assembled. The amino acid sequence requirements for the endopeptidase HycI involved in the C-terminal processing of HycE, the large subunit of the hydrogenase 3 from Escherichia coli, were investigated. Mutational alteration of the amino acid residues neighbouring the cleavage site showed that proteolysis still occurred when chemically similar amino acids were exchanged. Processing was blocked, however, in a variant in which the methionine at the C terminal side was replaced by a glutamate residue. Truncation of the precursor from the C-terminal end rendered variants amenable to maturation even when two thirds of the extension were removed but abolished proteolysis upon further deletion of a cluster of six basic amino acids. A construct in which the C terminal extension from the large subunit of the hydrogenase 2 was fused to the mature part of the large subunit of hydrogenase 3 was neither processed by HycI nor by HybD, the endopeptidase specific for the large subunit of hydrogenase 2. The maturation endopeptidase, therefore, exhibits a relaxed sequence constraint in recognition of its cleavage site and does not require the entire C-terminal extension. The results point to an interaction of the C-terminus with some domain of the large subunit, rendering a conformation amenable to recognition by the endopeptidase. PMID- 10795683 TI - Characterization of a new type of sulfite dehydrogenase from Paracoccus pantotrophus GB17. AB - The periplasmic sulfite dehydrogenase of Paracoccus pantotrophus GB17 was purified to homogeneity by a four-step procedure from cells grown lithoautotrophically with thiosulfate. The molecular mass of native sulfite dehydrogenase was 190 kDa as determined by native gradient PAGE. SDS-PAGE showed sulfite dehydrogenase to comprise two subunits with molecular masses of 47 kDa and 50 kDa, suggesting an alpha2beta2 structure. The N-terminal amino acid sequence and immunochemical analysis using SoxC-specific antibodies identified the 47-kDa protein as the soxC gene product. SoxD-specific antibodies identified the 50-kDa protein as SoxD. Based on the molecular masses deduced from the nucleotide sequence for mature SoxC (43,442 Da) and SoxD (37,637 Da) sulfite dehydrogenase contained 1.30 mol molybdenum/mol alpha2beta2 sulfite dehydrogenase. The iron content was 3.17 mol/mol alpha2beta2 sulfite dehydrogenase, and 3.53 mol heme/mol alpha2beta2 sulfite dehydrogenase was determined by pyridine hemochrome analysis. These data are consistent with the two heme-binding domains (CxxCH), characteristic for c-type cytochromes, deduced from the soxD nucleotide sequence. Electrospray ionization revealed two masses for SoxC of 43,503 and 43,897 Da. The difference in molecular mass was attributed to the molybdenum cofactor of SoxC. For SoxD a mass of 38,815 Da was determined; this accounted for the polypeptide and two covalently bound hemes. Reconstitution of the catalytic activity of sulfite dehydrogenase required additional fractions; these eluted from Q Sepharose at 0.05, 0.25, and 0.30 M NaCl. The K(m) of sulfite dehydrogenase for sulfite was 7.0 microM and for cytochrome c 19 microM. Sulfite dehydrogenase activity was inhibited by sulfate and phosphate. The structural and catalytic properties make sulfite dehydrogenase from P. denitrificans GB17 distinct from sulfite oxidases of other prokaryotic or eukaryotic sources. PMID- 10795684 TI - Characterization of abundance and diversity of lactic acid bacteria in the hindgut of wood- and soil-feeding termites by molecular and culture-dependent techniques. AB - Lactic acid bacteria have been identified as typical and numerically significant members of the gut microbiota of Reticulitermes flavipes and other wood-feeding lower termites. We found that also in the guts of the higher termites Nasutitermes arborum (wood-feeding), Thoracotermes macrothorax, and Anoplotermes pacificus (both soil-feeding), lactic acid bacteria represent the largest group of culturable carbohydrate-utilizing bacteria (3.6-5.2x10(4) bacteria per gut; 43%-54% of all colonies). All isolates were coccoid and phenotypically difficult to distinguish, but their enterobacterial repetitive intergenic consensus sequence (ERIC) fingerprint patterns showed a significant genetic diversity. Six different genotypes each were identified among the isolates from R. flavipes and T. macrothorax, and representative strains were selected for further characterization. By 16S rRNA gene sequence analysis, strain RfL6 from R. flavipes was classified as a close relative of Enterococcus faecalis, whereas strain RfLs4 from R. flavipes and strain TmLO5 from T. macrothorax were closely related to Lactococcus lactis. All strains consumed oxygen during growth on glucose and cellobiose; oxygen consumption of these and other isolates from both termite species was due to NADH and pyruvate oxidase activities, but did not result in H2O2 formation. In order to assess the significance of the isolates in the hindgut, denaturing gradient gel electrophoresis was used to compare the fingerprints of 16S rRNA genes in the bacterial community of R. flavipes with those of representative isolates. The major DNA band from the hindgut bacterial community was further separated by bisbenzimide-polyethylene glycol electrophoresis, and the two resulting bands were sequenced. Whereas one sequence belonged to a spirochete, the second sequence was closely related to the sequences of the Lactococcus strains RfLs4 and TmLO5. Apparently, those isolates represent strains of a new Lactococcus species which forms a significant fraction of the complex hindgut community of the lower termite R. flavipes and possibly also of other termites. PMID- 10795685 TI - Successional changes in the genetic diversity of a marine bacterial assemblage during confinement. AB - The successional changes in the genetic diversity of Mediterranean bacterioplankton subjected to confinement were studied in an experimental 300 1 seawater enclosure. Five samples were taken at different times and analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting to rapidly monitor changes in the bacterial genetic diversity. DGGE analysis clearly showed variations between the samples. Three of the five samples, with different DGGE banding patterns, were further analyzed by cloning and sequencing of 16S rRNA genes. Comparative sequence analysis indicated a shift from a mixed bacterial assemblage to a community dominated by bacteria closely affiliated to a single genus, Alteromonas. Sequences obtained at the start of the experiment were affiliated with two alpha-proteobacterial and three gamma proteobacterial lineages known from other studies of marine picoplankton. One sequence was affiliated with the Verrucomicrobiales. After 161 h of incubation two sequences represented a gamma-proteobacterial lineage also present at 0 h, but the majority of sequences clustered around that of Alteromonas macleodii. After 281 h only the dominant Alteromonas-like bacteria and bacteria distantly related to Legionella were found by cloning and sequencing. Mortality rates of bacteria indicated that grazing was the dominant mortality process when heterotrophic protozoa were abundant. Hence, changes in the genetic diversity of bacteria were partly influenced by the differential mortality of bacterial populations during the course of incubation. PMID- 10795687 TI - Unusual ultrastructural features in three strains of Cyanothece (cyanobacteria). AB - Three unicellular cyanobacterial strains (PCC 7425, PCC 8303, PCC 9308) assigned to the genus Cyanothece Komarek 1976, which showed an unusually high content of light refractile inclusions when viewed by phase-contrast microscopy, were characterized by confocal laser scanning microscopy and transmission electron microscopy. All strains had concentric cortical thylakoids and a compact central nucleoid. Frequently, the two innermost thylakoid membranes protruded to form circular enclosures containing cytoplasm or electron-transparent granules, or both. The largest granules were partially immersed in the nucleoid region, but they remained attached to the inner cortical thylakoids by a single narrow connection. The pattern of binary cell division in strain PCC 7425 was different than that in strains PCC 8303 and PCC 9308. In the former, all cell wall layers invaginated simultaneously, whereas in the latter the invagination of the outer membrane was delayed compared to that of the cytoplasmic membrane and the peptidoglycan layer. Thus, prior to completion of cell division, the new daughter cells of strains PCC 8303 and PCC 9308 were transiently connected by a thick septum, which was not observed in strain PCC 7425. Nucleoid partitioning coincided with initiation of cell division in all three strains and was unlike that reported in other bacteria and in archaea, in which separation of the nucleoids precedes cell division. Based on the common morphological and ultrastructural features, the three strains of Cyanothece examined constitute a distinct cluster, which might deserve independent generic status. PMID- 10795686 TI - Isolation and characterization of two new methanesulfonic acid-degrading bacterial isolates from a Portuguese soil sample. AB - Two novel bacterial strains that can utilize methanesulfonic acid as a source of carbon and energy were isolated from a soil sample collected in northern Portugal. Morphological, physiological, biochemical and molecular biological characterization of the two isolates indicate that strain P1 is a pink-pigmented facultative methylotroph belonging to the genus Methylobacterium, while strain P2 is a restricted methylotroph belonging to the genus Hyphomicrobium. Both strains are strictly aerobic, degrade methanesulfonate, and release small quantities of sulfite into the medium. Growth on methanesulfonate induces a specific polypeptide profile in each strain. This, together with the positive hybridization to a DNA probe that carries the msm genes of Methylosulfonomonas methylovora strain M2, strongly endorses the contention that a methanesulfonic acid monooxygenase related to that found in the previously known methanesulfonate utilizing bacteria is present in strains P1 and P2. The isolation of bacteria containing conserved msm genes from diverse environments and geographical locations supports the hypothesis that a common enzyme may be globally responsible for the oxidation of methanesulfonate by natural methylotrophic communities. PMID- 10795688 TI - Tc-99m MDP uptake in soft tissue extraskeletal metastasis from osteogenic sarcoma. AB - A bone scan in a patient with proved osteogenic sarcoma of the tibia showed intense focal uptake in the gluteal region on the side of his cancer. This was proved to be a metastasis in the muscle. PMID- 10795689 TI - Lymphoscintigraphy demonstrating thoracic duct injury in an infant with hypoplastic left heart syndrome. AB - Chylous effusion is an infrequent complication of cardiothoracic surgery and a less frequent complication of central line placement. The authors describe a novel application of lymphoscintigraphy showing thoracic duct injury in an infant after surgery for hypoplastic left heart syndrome. PMID- 10795690 TI - Hypophosphatemic osteomalacia demonstrated by Tc-99m MDP bone scan: a case report. AB - Hypophosphatemic osteomalacia, a familial or rarely acquired disorder, is characterized biochemically by hypophosphatemia, decreased renal tubular reabsorption of phosphate, decreased intestinal absorption of calcium, and normal serum calcium. This report concerns a rare case of hypophosphatemic osteomalacia of unknown cause that was shown on Tc-99m MDP bone scanning. PMID- 10795691 TI - The utility of thyroid nuclear imaging and other studies in the detection and treatment of underlying thyroid abnormalities in patients with endogenous subclinical thyrotoxicosis. AB - PURPOSE: Endogenous subclinical thyrotoxicosis is diagnosed when a patient who is not taking exogenous thyroid hormone has a suppressed level of thyroid stimulating hormone with normal levels of the free thyroid hormones thyroxine and triiodothyronine and other known causes of a suppressed thyroid-stimulating hormone level have been excluded. Although such a condition is caused by underlying thyroid disease, the specific nature and relative prevalence of these disorders and the utility of nuclear imaging and other studies in their detection remains unclear. PATIENTS AND METHODS: The authors performed a retrospective study of 50 patients with endogenous subclinical thyrotoxicosis. The results of the history and physical examination, thyroid nuclear scan, radioactive iodine uptake measurement, and thyroid antibody studies were reviewed. The results of the nuclear imaging and thyroid antibody studies were combined in an attempt to establish an underlying diagnosis for each patient. RESULTS: The thyroid nuclear imaging and antibody studies were used to establish a specific thyroid disorder in most of the patients (n = 39). These disorders included most commonly toxic multinodular goiter, various forms of autoimmune thyroid disease, and solitary toxic adenoma. A specific diagnosis was not determined in 11 patients. Therapy with I-131 radioactive iodine was administered to 14 of these patients, 13 of whom subsequently achieved a normal thyroid-stimulating hormone level. CONCLUSIONS: Most patients with endogenous subclinical thyrotoxicosis have underlying thyroid abnormalities that can be determined by nuclear imaging and, in selected cases, thyroid antibody studies. PMID- 10795692 TI - Recurrent hyperfunctioning parathyroid gland demonstrated on radionuclide imaging and an intraoperative gamma probe. AB - Radionuclide imaging with Tc-99m MIBI is the preferred mode of parathyroid localization in current practice. It also successfully identifies hyperfunctioning autotransplanted parathyroid tissue. The authors report a case with recurrent hyperparathyroidism after total parathyroidectomy and autotransplantation in the forearm. Double-phase Tc-99m MIBI imaging successfully localized the hyperfunctioning parathyroid tissue, which was missed by magnetic resonance imaging and ultrasonography. In addition, the parathyroid tissue was localized using an intraoperative probe at subsequent surgery. PMID- 10795693 TI - Tc-99m sestamibi before and during treatment in a patient with sarcoidosis and persistent hyperparathyroidism. AB - PURPOSE: Tc-99m sestamibi (MIBI) uptake in pulmonary sarcoidosis has been reported, but it has never been studied before and during treatment with glucocorticoids. METHODS: The authors performed MIBI scintigraphy and somatostatin receptor scintigraphy in a patient with sarcoidosis of the mediastinum, lungs, and liver and who had persistent hyperparathyroidism after unsuccessful neck exploration. RESULTS: Somatostatin receptor scintigraphy showed high mediastinal and pulmonary uptake in a pattern characteristic of sarcoidosis. Sustained MIBI uptake occurred in the same, although smaller, region. After the diagnosis was confirmed by liver biopsy, the patient was treated with glucocorticoids. Repeated MIBI scintigraphy showed that the uptake in the mediastinum had clearly decreased. CONCLUSIONS: This decrease of MIBI uptake in the mediastinal lymph nodes with therapy may imply that MIBI can be used to assess the response to treatment in sarcoidosis. Perhaps a relation exists between MIBI uptake at the moment of diagnosis and prognosis. Further studies in more patients are needed to evaluate the role of MIBI in the management of sarcoidosis. PMID- 10795694 TI - Use of a reusable shielded marker to enhance the accuracy, safety, and efficacy of nuclear medicine procedures. AB - Three cases illustrate the use of a reusable, shielded marker to identify anatomic structures and mark pathologic lesions. No other nuclear medicine marker is available with a shutter mechanism designed to interrupt radiation, thus protecting the patient and technologist from unnecessary radiation and minimizing image artifacts. PMID- 10795695 TI - Standardized uptake value as an unreliable index of renal disease on fluorodeoxyglucose PET imaging. AB - Fluorine-18 2-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography has been used extensively in the diagnosis of malignant conditions with high rates of sensitivity and specificity. However, increased FDG uptake is not limited to malignant tissue. In general, lesions with a mild degree of FDG uptake as measured by standardized uptake values less than 2.0 are considered benign, whereas those with values greater than 2.5 are usually regarded as malignant. Standardized uptake values in the kidney can be as high as 22 as a result of excretion of FDG through urine. Two cases are reported in which renal abnormalities could not be distinguished from urine based on standard uptake values alone. PMID- 10795696 TI - Abnormal pulmonary accumulation of indium-111 chloride in pneumocystis carinii pneumonia as detected by bone marrow scintigraphy. AB - PURPOSE: Unusual pulmonary uptake of In-111 chloride in a patient with Pneumocystis carinii pneumonia and autoimmune hepatitis is described. METHOD: In 111 chloride bone marrow scintigraphy was performed to evaluate the bone marrow activity associated with pancytopenia in a 56-year-old woman with autoimmune hepatitis. RESULTS: An In-111 chloride bone marrow scan showed increased pulmonary uptake predominantly in both upper lung fields. P. carinii pneumonia was seen to be developing as an immunocompromised complication after treatment for autoimmune hepatitis. CONCLUSION: When In-111 chloride bone marrow scintigraphy shows increased uptake in the lungs of immunocompromised patients, a combined opportunistic inflammatory disease such as P. carinii pneumonia should be considered in the diagnosis. PMID- 10795697 TI - Comparative study of Tc-99m MIBI and TI-201 SPECT in predicting chemotherapeutic response in non-small-cell lung cancer. AB - PURPOSE: Tc-99m MIBI can be excluded from cytosol against its concentration gradient as a suitable transport substrate by P-glycoprotein. Tc-99m MIBI has also been shown to be more effective than TI-201 chloride for evaluating the response to chemotherapy in patients with small-cell lung cancer. The relation between Tc-99m MIBI accumulation by the tumor and its response to chemotherapy were evaluated in patients with non-small-cell lung cancer (NSCLC) and compared with the same parameters achieved using TI-201 chloride. METHODS: Thirty-eight patients with NSCLC were examined before chemotherapy was begun. They were classified according to the results of a follow-up computed tomogram into two groups: responders were patients in whom there was a > or =50% decrease and nonresponders were patients in whom there was a <50% decrease in the sum of the product of the maximum perpendicular diameters of all measurable lesions. All patients underwent dual-isotope imaging with TI-201 chloride and Tc-99m MIBI just before chemotherapy. Regions of interest were placed over the tumor uptake (T) and contralateral normal lung tissue (N) areas on one coronal view with a clearly defined lesion, and the T:N ratio and retention index were calculated. RESULTS: The delayed T:N ratio and retention index for Tc-99m MIBI in the responder group were significantly greater (P<0.05) than those in the nonresponder group. There was no significant correlation between the T:N ratio and retention index and tumor response using TI-201 chloride. CONCLUSION: Tc-99m MIBI SPECT may be more effective than TI-201 chloride SPECT for evaluating the response to chemotherapy in patients with NSCLC. PMID- 10795698 TI - Detection of synchronous carcinomas of the colon with F-18 fluorodeoxyglucose: a case report. AB - Colon and rectal carcinomas are common in North America and Northwestern Europe. In South America, the risk of this disease is not as great. Adenocarcinomas are the most common types of tumors, and they occur mainly in the descending colon, sigmoid, and rectum. A patient with a possible right colon carcinoma was examined using F-18 FDG. PMID- 10795699 TI - Avascular necrosis and fracture of the capitate bone: unusual scintigraphic features. PMID- 10795700 TI - Positive Tc-99m MIBI breast study related to a psoriatic lesion. PMID- 10795701 TI - Iodine-131 uptake in thymic hyperplasia with atypical computed tomographic features. PMID- 10795702 TI - Malignant supraclavicular lymph node visualization during Tc-99m HDP bone imaging. PMID- 10795703 TI - Heart and lung accumulation of Tc-99m MDP with normal radiographs in patients undergoing hemodialysis. PMID- 10795704 TI - Prominent imaging of the liver on bone densitometry: metastasis of papillary thyroid carcinoma. PMID- 10795705 TI - Demonstration of renal lithiasis on technetium-99m MDP bone scintigraphy. PMID- 10795706 TI - Ga-67 scintigram in the diagnosis of infection of masticator muscles due to an odontogenic infection. PMID- 10795707 TI - Thymoma with pure red cell aplasia demonstrated by Tc-99m tetrofosmin SPECT. PMID- 10795708 TI - Tc-99m MDP uptake in herniated stomach tissue. PMID- 10795709 TI - A case of acute tubular necrosis with intestinal visualization of Tc-99m DMSA. PMID- 10795710 TI - Ga-67 scintigraphy in a patient with B-cell lymphoma. PMID- 10795711 TI - An unusual case of unilateral lactation and a vanishing limb. PMID- 10795712 TI - Sunburst periosteal reaction in a bony metastasis. PMID- 10795713 TI - Evaluation of adrenocortical carcinoma in an infant using Ga-67 scintigraphy. PMID- 10795714 TI - Current readings in nuclear medicine. PMID- 10795715 TI - Principles of animal use for gerontological research. AB - This essay presents some practical advice and suggestions for those who wish to use mice and rats in experiments on the biology of aging. Ten principles set forth guidance on choice of ages, choice of stocks, the importance of specific pathogen-free status, the uses of necropsy data, the dangers of pooling samples from different individuals, planning ahead for loss of aged mice to death and disease, the use of cost-adjusted power calculations, and the dangers of inferring causal associations from correlated age effects. PMID- 10795716 TI - Age-related decline in Ras/ERK mitogen-activated protein kinase cascade is linked to a reduced association between Shc and EGF receptor. AB - Numerous studies have demonstrated that the proliferative capacity of cells declines with age. Using rat primary hepatocytes as a model system, we recently demonstrated that this age-related decline in the proliferative response to mitogenic stimulation is associated with decreased activities of both extracellular signal-regulated kinase (ERK) and p70 S6 kinase (p70(S6k)). To unravel the molecular basis for age-related defects in the ERK pathway, we have now characterized the upstream signaling events that occur after epidermal growth factor (EGF) stimulation in young and aged hepatocytes. As previously noted for ERK, the activities of both MEK (the kinase immediately upstream of ERK) and Ras following EGF stimulation were significantly lower in aged hepatocytes. An examination of the EGF receptor (EGFR) revealed a similar amount of EGFR in the two age groups. Likewise, EGFR and Shc, an adaptor protein that plays a crucial role in linking EGFR to Ras activation, underwent tyrosine phosphorylation to a similar degree in both young and aged hepatocytes. However, in aged cells Shc was unable to form stable complexes with EGFR after EGF stimulation. Our results suggest that a decrease in the association between Shc and EGFR in aged cells underlies the age-related declines in the ERK signaling cascade and in proliferative capacity. PMID- 10795717 TI - Age-dependent increase in infarct volume following photochemically induced cerebral infarction: putative role of astroglia. AB - This study demonstrates that the photochemically induced model of stroke is an extremely viable method of inducing cerebral infarction in old animals. The lesions are reproducible both in terms of location and size and compatible with long-term survival of the animal. With this model we demonstrated, one week following surgery, a significantly larger infarct in rats 20 and 24 months of age compared to 4-month-old rats. The older rats also sustained greater neurologic deficits as assessed on a rotarod task. Older rats also were characterized by a glial response that was far less intense than in young animals. While the precise relationship between glia activation and cerebral damage remains to be determined, it would appear that a better understanding of those factors that contribute to the astrocytic response in the aged rat may be of particular benefit in designing therapeutic strategies aimed at reducing the pathologic consequences of cerebral infarction in elderly humans. PMID- 10795718 TI - Effect of fruits, vegetables, or vitamin E--rich diet on vitamins E and C distribution in peripheral and brain tissues: implications for brain function. AB - Age-related neurodegenerative conditions are the principal cause of declining cognitive and motor function during aging. Evidence support that fruits and vegetables containing generous amounts of antioxidant nutrients are important for neurological function. We investigated the effect of diets enriched with fruits or vegetables but low in vitamin E and a diet high in vitamin E on the distribution of vitamins C and E in the brain and dopamine release of Fischer 344 rat model, over an 8-month period. The low-vitamin E diet resulted in lowered alpha-tocopherol levels in brain and peripheral tissues, whereas the animals that received a diet enriched in vitamin E showed a significant increase, between 500 900%. Vitamin C concentration in plasma, heart, and liver was reduced in the vitamin E-supplemented group. It is concluded that supplementation or depletion of alpha-tocopherol for 8 months results in marked changes in vitamin E levels in brain tissue and peripheral tissues, and varied distribution of alpha-tocopherol throughout the different brain regions examined. In addition, compared to control group, rats supplemented with strawberry, spinach, or vitamin E showed a significant enhancement in striatal dopamine release. These findings suggest that other nutrients present in fruits and vegetables, in addition to the well-known antioxidants, may be important for brain function. PMID- 10795719 TI - Effects of strength training and detraining on muscle quality: age and gender comparisons. AB - Maximal force production per unit of muscle mass (muscle quality, or MQ) has been used to describe the relative contribution of non-muscle-mass components to the changes in strength with age and strength training (ST). To compare the influence of age and gender on MQ response to ST and detraining, 11 young men (20-30 years), nine young women (20-30 years), 11 older men (65-75 years), and 11 older women (65-75 years), were assessed for quadriceps MQ at baseline, after 9 weeks of ST, and after 31 weeks of detraining. MQ was calculated by dividing quadriceps one repetition maximum (IRM) strength by quadriceps muscle volume determined by magnetic resonance imaging. All groups demonstrated significant increases in IRM strength and muscle volume after training (all p < .05). All groups also increased their MQ with training (all p < .01), but the gain in MQ was significantly greater in young women than in the other three groups (p < .05). After 31 weeks of detraining, MQ values remained significantly elevated above baseline levels in all groups (p < .05), except the older women. These results indicate that factors other than muscle mass contribute to strength gains with ST in young and older men and women, but those other factors may account for a higher portion of the strength gains in young women. These factors continue to maintain strength levels above baseline for up to 31 weeks after cessation of training in young men and women, and in older men. PMID- 10795720 TI - Glyceraldehyde-3-phosphate dehydrogenase varies with age in glycolytic muscles of rats. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, protein, and enzyme activity levels in hindlimb muscles of adult and senescent Fischer 344 x Brown Norway rats were investigated. Soleus muscles from adult and senescent rats had similar levels of GAPDH. In contrast, muscles containing a large proportion of glycolytic fibers had lower GAPDH levels in senescent rats relative to these muscles in adult rats; this was observed at both the mRNA and protein levels. These data indicate that skeletal muscle glycolytic capacity of fast muscles is diminished with age and that it may be caused by changes at the level of transcription. Also, because GAPDH mRNA levels change with age in several rat muscles, GAPDH mRNA is not always a proper internal control for mRNA analyses of aging skeletal muscle. PMID- 10795721 TI - Cognitive influence on postural stability: a neuromuscular analysis in young and older adults. AB - BACKGROUND: Previous literature indicates that attentional resources are required for recovery of postural stability. Previous studies have also examined the effect of aging on the performance of a static postural task while a secondary cognitive task is being conducted. This study describes the effect of a cognitive task on the neuromuscular response characteristics underlying reactive balance control in young versus older adults. METHODS: The attentional demand on the neuromuscular system was examined in 14 young and 12 healthy older adults by analysis of the integrated electromyography activity while the adults were performing a dual-task paradigm. The primary task involved standing platform perturbations and the secondary task was a math task that involved subtraction by threes. Integrated electromyography activity was compared between the cognitive (math and balance) task versus control (balance only) task. RESULTS: For both groups of subjects, onset latency of postural muscle responses did not change under dual-task conditions. In contrast, the amplitude of postural muscle activity was significantly affected by performance of a secondary task. When electromyography data were combined for both young and older adults, there was a decrease in muscle response amplitude in both agonist (gastrocnemius) and antagonist (tibialis anterior) muscles when the cognitive math task was performed. This was apparent at 350-500 milliseconds from plate onset for the gastrocnemius and between 150 and 500 for the tibialis anterior. When young and older adults were compared, an age by task interaction effect was seen in muscle response amplitude for the agonist (gastrocnemius) muscle between 350 to 500 milliseconds, with older adults showing a significantly greater reduction than young adults. CONCLUSION: The decline of muscle activity when the secondary task was performed suggests that less attentional processing capacity was available for balance control during the dual-task paradigm. The results also indicate that the dual-task activity has a greater impact on balance control in the older adults than in the young adults. PMID- 10795722 TI - Sleep-wake patterns among postmenopausal women: a 24-hour unattended polysomnographic study. AB - BACKGROUND: Circadian sleep-wake profiles in postmenopausal women were examined to explore relationships between nocturnal and out-of-bed sleep. METHODS: Twenty one home recordings were obtained with unattended polysomnography from women ranging from 56 to 77 years of age. RESULTS: While maintaining their daily routines, volunteers slept an average of 439 minutes throughout the 24-hour recordings. Ten percent of the accumulated sleep time was recorded out of bed. CONCLUSIONS: Greater age was associated with more afternoon-evening sleep. Sleep was also frequently observed shortly after volunteers arose from bed in the morning. PMID- 10795723 TI - "Warm-up" phenomenon in adult and elderly patients with coronary artery disease: further evidence of the loss of "ischemic preconditioning" in the aging heart. AB - BACKGROUND: A reduction of exercise-induced ischemia in patients with coronary artery disease by means of brief period of exercise followed by resting is called the "warm-up" phenomenon. This phenomenon may represent a clinical counterpart of "ischemic preconditioning." We studied the warm-up phenomenon in both adult and elderly patients with similar angiographic evidence of coronary artery disease, using three exercise tests after excluding the "training effect." METHODS: In order to verify the presence of "training effect," three exercise tests were performed in days 1, 2, and 3 ("training" tests). The third test was used as baseline for a successive test, performed after a recovery period of 10 minutes to reestablish baseline electrocardiographic conditions. A third exercise test was performed 30 minutes later ("warm-up" tests). RESULTS: "Training" tests did not differ in all parameters in both adult and elderly patients. "Warm-up" tests showed that time to onset 1-mm ST depression was significantly higher (p < .001). whereas ST depression and time to recovery was significantly lower in the second and third test in adult but not in elderly patients (p < .001 ). Difference (in seconds) in the time at which 1-mm ST depression occurred on first warm-up exercise compared with the second was inversely correlated with age (p < .001 ). CONCLUSIONS: Previous exercise followed by resting is able to reduce the successive exercise-induced ischemia ("warm-up" phenomenon) in adult but not in elderly patients with coronary artery disease. This is independent of a greater age-related severity of coronary disease and of "training effect." These results confirm the hypothetical age-related reduction of "ischemic preconditioning" in aging heart. PMID- 10795724 TI - The cross-sectional association between blood pressure and Alzheimer's disease in a biracial community population of older persons. AB - BACKGROUND: The relation of blood pressure to Alzheimer's disease (AD) is complex because both an association of high blood pressure with increased risk of the disease and lower blood pressure as a consequence of the disease are possible. METHODS: We examined the cross-sectional association of blood pressure and AD in the Chicago Health and Aging Project (CHAP), a study of a geographically defined, biracial community. After in-home interviews with 6.162 residents > or =65 years, a stratified random sample of 729 participants was clinically evaluated; 709 had blood pressures measured, and 243 were diagnosed with AD. RESULTS: In logistic regression models adjusted for age, sex, education, and race there was no association between blood pressure measured as a continuous variable and Alzheimer's disease. In categorical analyses, however, prevalence of Alzheimer's disease was significantly higher among persons with low systolic pressure (<130 mmHg) compared with the referent group of 130-139 mmHg (odds ratio [OR] = 2.2, 95% confidence interval [CI]: 1.2,4.1), and with low diastolic pressure (<70 mmHg) compared to the referent of 70-79 mmHg (OR = 1.8, 95% CI: I. 1,3.1). High systolic and diastolic categories were not statistically different from the referent group, although there was some evidence that the associations differed by race. The odds ratios changed little with further adjustment for apolipoprotein E genotype, antihypertensive medications, body mass, stroke, diabetes, and heart disease. CONCLUSION: These findings are consistent with previous studies showing associations between low blood pressure and AD, but longitudinal studies are needed to characterize cause-and-effect associations. PMID- 10795725 TI - Effect of age and menopause on serum concentrations of pentosidine, an advanced glycation end product. AB - BACKGROUND: Pentosidine is an advanced glycation end product. Our aim is to investigate (a) the age-related change of serum pentosidine and (b) the effect of menopause on serum pentosidine. METHODS: Using the high-performance liquid chromatography method with column switching, we measured serum pentosidine in 140 healthy women aged 20-93 years. Serum creatinine was also measured. The samples of 13 young and 13 old subjects were used for the measurements of free pentosidine and fractions of pentosidine. Free pentosidine was measured without hydrolysis, and the fractions were measured with a 10,000 mol wt cutoff filter. To investigate the effect of menopause on pentosidine, two biochemical markers for bone turnover (CTx and osteocalcin) were measured in age-matched premenopausal and postmenopausal women (16 in each group). RESULTS: Serum pentosidine significantly increased with age (r = .702, p < .0001 ). The values of serum pentosidine for the groups beyond the age of 50 were significantly higher than those for the younger groups. The value for the group aged 80-93 years was three times higher than that for the group aged 20-29 years. Serum pentosidine moderately and significantly correlated to serum creatinine (r = .483, p = .0001). Free pentosidine was detected in only 3 of 13 young subjects and 2 of 13 old subjects. The ratio of free to total pentosidine was 2.9% and 1.2% in young and old subjects, respectively. Pentosidine <10,000 mol wt was not detected in all subjects. Pentosidine >10,000 mol wt was detected in all subjects. Serum CTx and osteocalcin significantly increased in postmenopausal women compared with those of pre-menopausal women. There was no significant change in serum pentosidine between the premenopause group and the postmenopause group. CONCLUSION: Serum pentosidine significantly increased with age in healthy subjects aged 20-93 years and correlated to serum creatinine. The changes of fractions of pentosidine with aging were not observed. There was no effect of menopause on pentosidine. PMID- 10795726 TI - Maintenance of bone density: outcomes over five years of attendance at an osteoporosis center. AB - BACKGROUND: This is a longitudinal analysis of the data from women who visited the Osteoporosis Prevention Center for at least five years. The study is part of an ongoing evaluation of the results of the Center visits. METHODS: A random sample of 100 women who had attended the clinic for more than five years and were not referred for a definite medical diagnosis was pulled from the records. Descriptive statistics were compiled on this sample. Seventy-five women were postmenopausal and over 49 years of age. This subgroup was studied for change in bone mineral density (BMD) over the five years. RESULTS: The BMD change was 0.03 gm/cm2 in the spine, -0.01 gm/cm2 in the femoral-neck. and -0.008 gm/cm2 at the radius over the five years. CONCLUSION: Attendance at the Osteoporosis Prevention Center was associated with maintenance of bone density in the spine over a five year period. PMID- 10795727 TI - Gait characteristics of young and older individuals negotiating a raised surface: implications for the prevention of falls. AB - BACKGROUND: Falls in older individuals are a major public health issue because of the financial cost of surgery and re habilitation and the human cost of associated pain and disability. Older individuals are most likely to fall when negotiating an obstacle or obstruction during locomotion. This research was aimed at investigating lower limb motion while a subject negotiated a raised surface. METHODS: The gait of six healthy young (Y) women (mean age 23.1 years) and six healthy older (O) women (mean age 67.6 years) were analyzed with a PEAK motion analyzer and a dual-force-platform system during unobstructed walking and when the subjects were stepping on and off a raised surface of 15 cm. The effect of age on foot clearance and force platform variables was analyzed. RESULTS: During stepping on, the young women cleared the step by the lead foot by a significantly greater margin than the older subjects did (Y = 10.6 cm, O = 9.1 cm; p < .05) but trail-foot clearance was not significantly different (Y 9.4 cm, O = 8.8 cm). Foot clearance in stepping off was low compared with that of ascent, and the older individuals had a significantly higher lead (Y = 1.5 cm, O = 3.3 cm, p < .05) and trail (Y = 1.0 cm, O = 2.1 cm) vertical clearance. Older individuals positioned both the lead and the trail foot relatively farther from the step edge on ascending a raised surface, respectively, Y = 87% and O = 93% of the step cycle and Y = 29% and O = 34%. Foot placement in descent was qualitatively similar for the two groups. The force and the impulse data under the lead and the trail feet confirm modulations consistent with the foot clearance data. CONCLUSION: In negotiating a raised surface older individuals appear to use a nonoptimal foot placement strategy in which, compared with that of young subjects, the trail foot is placed a long way from the edge of the step. The older subjects allowed very little correction time and little latitude in foot placement beyond the edge of the step, suggesting that the approach to the obstacle may be a critical determinant of safety. PMID- 10795728 TI - Increased plasma norepinephrine response to yohimbine in elderly men. AB - BACKGROUND: The effects of aging on sympathetic nervous system and adrenomedullary outflow were estimated by the measurement of plasma norepinephrine (NE) and epinephrine (EPI) responses to yohimbine and clonidine in healthy young and healthy older subjects. METHODS: Yohimbine (0.65 mg/kg), clonidine (5 microg/kg), and placebo were administered on separate days in random order to 5 healthy older men (age 74 +/- 1 years) and 18 healthy young men (age 26 +/- 1 years). NE and EPI were measured by radioenzymatic assay in plasma samples obtained before and 30, 60, and 90 minutes after drug administration. RESULTS: Plasma NE increases after yohimbine were greater in older men than in young men. but plasma NE decreases following clonidine did not differ between groups. Plasma NE and systolic blood pressure were higher in older men than in young men at baseline but no longer differed 90 minutes after clonidine. Plasma EPI increases after yohimbine and decreases after clonidine did not differ between groups. CONCLUSIONS: These results suggest increased sympathetic nervous system outflow in human aging that is not a function of reduced responsiveness of alpha-2 adrenoreceptor-mediated feedback inhibition. PMID- 10795729 TI - Postmenopausal hormone replacement therapy and cardiovascular mortality in Central-Eastern Europe. AB - BACKGROUND: The leading cause of death among elderly women is cardiovascular (CV) disease in the United States and in Western Europe as well. The protective effect of postmenopausal hormone replacement therapy (HRT) on coronary heart disease has been verified in epidemiologic studies. There are no data available on the rate of HRT use in Eastern Europe. Our goals were to study the rates of HRT in Eastern Europe, to compare them to those of the United States and Western Europe, as well as to compare their CV mortality rates. METHODS: The use of HRT in Eastern Europe was calculated from sales records obtained from all pharmaceutical companies that ship HRT preparations to the given area. Data on HRT in Western countries were taken from the literature. Mortality rates were obtained from the World Health Organization. RESULTS: The rate (mean +/- SD) of HRT in Eastern Europe was 2.88 +/- 2.67%, whereas 12.67 +/- 9.97% in Western Europe and the United States, p < .05. The cardiovascular mortality rate per 100,000 women older than 45 years in Eastern Europe was higher (1766 +/- 158.3) than in the Western countries (1155 +/ 164.1, p < .001). CONCLUSIONS: The rate of HRT is markedly lower. whereas CV mortality rates are notably higher in Eastern Europe than in the United States or Western Europe. Because HRT seems to be underutilized in Eastern Europe, to increase its use might be an important tool to improve CV mortality rates. However, due to the risks associated with HRT, other measures to prevent coronary heart disease, such as smoking cessation programs, and other efforts should also be considered in Eastern Europe. PMID- 10795730 TI - Noninvasive diagnosis of Helicobacter pylori infection in older subjects: comparison of the 13C-urea breath test with serology. AB - BACKGROUND: The potential influence of cognitive status, physical activities, comorbidity and cotreatments on the feasibility and diagnostic accuracy of two noninvasive diagnostic tests for Helicobacter pylori (Hp) infection, i.e., the 13C-urea breath test (13C-UBT) and serology (immunoglobulin G [IgG] anti-Hp antibodies), in older subjects is not known. METHOD: The study involved 100 consecutive symptomatic elderly subjects (mean age, 78.3 years; range, 65-96 years), who had undergone an upper gastrointestinal endoscopy. Patients were considered Hp positive if at least two of the three invasive methods, i.e. histology, culture, and/or the rapid urease test were positive for Hp infection. Patients were considered Hp negative if all three invasive methods were negative. The 13C-UBT was performed according to the European standard method and the assaying of IgG anti-Hp antibodies by enzyme-linked immunosorbent assay. Cognitive status and functional activities were determined by the Mini-Mental State Examination (MMSE), the activities of daily living (ADLs) and instrumental ADLs (IADLs). RESULTS: According to invasive methods, 49 patients were Hp positive and 47 were Hp negative (4 subjects were excluded from the study). Hp positive patients demonstrated a significantly higher prevalence of peptic ulcers (p =.02) and activity of chronic gastritis (p<.0001) than Hp-negative subjects. The 13C-UBT demonstrated a sensitivity of 100%, a specificity of 95.7%, and a diagnostic accuracy of 97.9%. Serology had significantly lower sensitivity (74.4%), specificity (59%), and diagnostic accuracy (67%, p<.001) than the 13C UBT. The feasibility and the diagnostic accuracy of the 13C-UBT were not altered by the cognitive status (MMSE) and functional activities (ADL, IADL) of the patients, their drug consumption, or the prevalence of concomitant diseases. CONCLUSIONS: In older subjects, the 13C-UBT had a significantly higher diagnostic accuracy than serology without influence of cognitive function, disability, comorbidity and cotreatments. This method may be considered an excellent, clinically useful, noninvasive test for the diagnosis of Hp infection in older subjects. PMID- 10795731 TI - Muscle strength and body mass index as long-term predictors of mortality in initially healthy men. AB - BACKGROUND: Muscle weakness, low body weight, and chronic diseases are often observed in the same people; however, the association of muscle strength with mortality, independent of disease status and body weight, has not been elucidated. The aim was to assess hand grip strength as a predictor of all-cause mortality within different levels of body mass index (BMI) in initially disease free men. METHODS: Mortality was followed prospectively over 30 years. Maximal hand grip strength tests and BMI assessments were done at baseline in 1965 to 1970. The participants were 6040 healthy men aged 45 to 68 years at baseline living on Oahu, Hawaii. RESULTS: The death rates per 1000 person years were 24.6 in those with BMI <20, 18.5 in the middle BMI category, and 18.0 in those with BMI > or = 25. For grip strength tertiles, the mortality rates were 24.8 in the lowest, 18.5 in the middle, and 14.0 in the highest third. In Cox regression models, within each tertile of grip strength, BMI showed only minimal effect on mortality. In contrast, in each category of BMI there was a gradient of decreasing mortality risk with increasing grip strength. Among those with BMI <20, the adjusted relative risks (RRs) of mortality over 30 years were 1.36 (95% confidence interval 1.14-1.63) for those in the lowest third of strength at baseline, 1.27 (1.02-1.58) in the middle, and 0.92 (0.66-1.29) in the highest third. Correspondingly, for those with BMI 20-24.99, the RRs of death were 1.25 (1.08-1.45), 1.14 (1.00-1.32), and 1.0 (reference) in the lowest, middle, and highest third of grip strength, respectively. In those with BMI > or =25, the RRs were 1.39 (1.16-1.65) in the lowest, 1.27 (1.08-1.49) in the middle, and 1.14 (0.98-1.32) in the highest third of grip strength. Models were adjusted for age, education, occupation, smoking, physical activity, and body height. CONCLUSIONS: In healthy middle-aged men, long-term mortality risk was associated with grip strength at baseline, independent of BMI. The possible interpretation of the finding is that early life influences on muscle strength may have long-term implications for mortality. Additionally, higher strength itself may provide greater physiologic and functional reserve that protects against mortality. PMID- 10795733 TI - Body composition prediction equations for elderly men. AB - BACKGROUND: The purpose of this study was to develop and validate anthropometric body composition prediction equations for elderly (i.e., > or =65 years old) men. This was necessary because of a lack of accurate and reliable predictive equations specifically developed for this population. METHODS: Seventy-five elderly men were randomly assigned to either an equation development sample (on = 50) or an equation validation sample (n = 25). Subject anthropometric measures were analyzed in a regression procedure with hydrodensitometry-determined body density, percentage of fat, fat-free mass, and fat weight to develop prediction equations for each body composition variable. The equation estimates were then validated against the hydrostatically determined measures. RESULTS: Four equations were developed and validated for the estimation of elderly male body composition variables [one each for body density (R2 = .66, SEE = +/- .01, where SEE is the standard error of estimate), percentage of fat (R2 = .66, SEE = +/- 4.43), fat-free mass (R2 = .88, SEE = +/- 3.94, and fat weight (R2 = .90, SEE = +/- 4.11)]. The equations provided estimates of body density, percentage of fat, fat-free mass, and fat weight, which were not statistically different from the hydrostatically determined criterion variables. CONCLUSIONS: The results of this study indicate that accurate and reliable anthropometric predictive equations can be developed for an active and healthy elderly male population. These equations may be used for accurate epidemiological testing of this group's body composition variables. PMID- 10795732 TI - Estrogen increases hyperemic microvascular blood flow velocity in postmenopausal women. AB - BACKGROUND: Epidemiologic studies suggest that estrogen replacement therapy (ERT) is protective against vascular disease. ERT confers this benefit by lowering lipid levels and improving arterial function. However, its effect on the microvasculature in vivo is unknown. Thus the purposes of this study were to evaluate effect of estrogen status on the hyperemic response of the microvasculature in vivo in postmenopausal women and to compare the hyperemic response of the microvasculature in postmenopausal women taking ERT with that of premenopausal women. METHODS: We measured forearm microvasculature flow velocity by using a laser Doppler in a cross section of 64 healthy premenopausal and postmenopausal women 23 to 72 years old. Microvasculature blood flow velocity was measured at baseline. throughout 2 minutes of ischemia, and immediately after the ischemic period was terminated (i.e., during the peak hyperemic response). RESULTS: The peak of the hyperemic flow velocity (PHFV) in the postmenopausal women who were taking long-term ERT at usual doses was greater than that of postmenopausal women who were not currently taking ERT (p < .0001). Moreover, the PHFV of postmenopausal women taking ERT was similar to that of premenopausal women. Multivariate regression analysis showed estrogen status and baseline flow velocity to be independent predictors of PHFV. CONCLUSIONS: Current, long-term ERT at usual replacement doses is associated with improved microvascular responses in postmenopausal women, which may explain some of its beneficial vascular effects. PMID- 10795734 TI - Manipulating immune responses with immunosuppressive agents that target NFAT. PMID- 10795735 TI - Tec kinases: a family with multiple roles in immunity. PMID- 10795736 TI - Highly efficient selection of CD4 and CD8 lineage thymocytes supports an instructive model of lineage commitment. AB - We undertook a kinetic analysis of the generation of mature T cells in TCR and coreceptor transgenic mice using BrdU labeling. We observed that the selection efficiency of mature CD4-CD8+ and CD4+CD8- thymocytes could be as high as 40% and 90% of CD4+CD8+ precursors, respectively. The surprisingly high efficiency of selection favors an instructional model of lineage commitment and is incompatible with a stochastic model in which the efficiency of selection would be no greater than 100% in both lineages combined. PMID- 10795737 TI - Control of antigen presentation by a single protease cleavage site. AB - Protein antigens require limited proteolytic processing to generate peptides for binding to class II MHC molecules, but the proteases and processing sites involved are largely unknown. Here we analyze the effect of eliminating the three major asparagine endopeptidase (AEP)-processing sites in the microbial antigen tetanus toxin C fragment. The mutant antigen is highly resistant to proteolysis by AEP and crude lysosomal extracts and is dramatically impaired in its ability to be processed and presented to T cells. Remarkably, processing at a single asparagine residue (1219) is obligatory for optimal presentation of many T cell epitopes in this antigen. These studies demonstrate that cleavage at a single processing site can be crucial for effective antigen presentation. PMID- 10795738 TI - HPK1 is activated by lymphocyte antigen receptors and negatively regulates AP-1. AB - The serine/threonine kinase HPK1 is a member of the germinal center kinase (GCK) family that has been implicated in the regulation of MAP kinase pathways. Here, we demonstrate the involvement of HPK1 in antigen receptor signaling. Engagement of the TCR or the BCR resulted in a marked induction of HPK1 catalytic activity. Subsequent analysis revealed that Src and Syk/ZAP-70 tyrosine kinases and the adaptor proteins LAT, SLP-76, BLNK, Grb2, and Grap are involved in HPK1 activation. Overexpression of HPK1 inhibited TCR activation of AP-1 and ERK2, whereas the kinase-inactive mutant of HPK1 potentiated these responses. Neither form of HPK1 affected PMA or v-Ras activation of AP-1 and ERK2. Thus, HPK1 is a negative regulator of the TCR-induced AP-1 response pathway. PMID- 10795739 TI - Role of CD8beta domains in CD8 coreceptor function: importance for MHC I binding, signaling, and positive selection of CD8+ T cells in the thymus. AB - The contribution of the CD8beta subunit to CD8 coreceptor function is poorly understood. We now demonstrate that the CD8beta extracellular domain increases the avidity of CD8 binding to MHC I, and that the intracellular domain of CD8beta enhances association with two intracellular molecules required for TCR signal transduction, Lck and LAT. By assessing CD8+ T cell differentiation in CD8beta deficient mice reconstituted with various transgenic CD8beta chimeric molecules, we also demonstrate that the intracellular and extracellular domains of CD8beta can contribute independently to CD8+ T cell development, but that both CD8beta domains together are most efficient. Thus, this study identifies the molecular functions of the CD8beta intracellular and extracellular domains and documents their contributions to CD8+ T cell development. PMID- 10795740 TI - The distinct roles of TRAF2 and RIP in IKK activation by TNF-R1: TRAF2 recruits IKK to TNF-R1 while RIP mediates IKK activation. AB - The death domain kinase RIP and the TNF receptor-associated factor 2 (TRAF2) are essential effectors in TNF signaling. To understand the mechanism by which RIP and TRAF2 regulate TNF-induced activation of the transcription factor NF-kappaB, we investigated their respective roles in TNF-R1-mediated IKK activation using both RIP-/- and TRAF2-/- fibroblasts. We found that TNF-R1-mediated IKK activation requires both RIP and TRAF2 proteins. Although TRAF2 or RIP can be independently recruited to the TNF-R1 complex, neither one of them alone is capable of transducing the TNF signal that leads to IKK activation. Moreover, we demonstrated that IKK is recruited to the TNF-R1 complex through TRAF2 upon TNF treatment and that IKK activation requires the presence of RIP in the same complex. PMID- 10795741 TI - B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes. AB - CD28/B7 costimulation has been implicated in the induction and progression of autoimmune diseases. Experimentally induced models of autoimmunity have been shown to be prevented or reduced in intensity in mice rendered deficient for CD28 costimulation. In sharp contrast, spontaneous diabetes is exacerbated in both B7 1/B7-2-deficient and CD28-deficient NOD mice. These mice present a profound decrease of the immunoregulatory CD4+CD25+ T cells, which control diabetes in prediabetic NOD mice. These cells are absent from both CD28KO and B7-1/B7-2KO mice, and the transfer of this regulatory T cell subset from control NOD animals into CD28-deficient animals can delay/prevent diabetes. The results suggest that the CD28/ B7 costimulatory pathway is essential for the development and homeostasis of regulatory T cells that control spontaneous autoimmune diseases. PMID- 10795743 TI - Returning control of health care to the patient-physician nexus. PMID- 10795742 TI - T cell progenitors emerge earlier than B cell progenitors in the murine fetal liver. AB - The developmental potential of individual cells in the Lin-c-kit+CD45+IL-7R+ (IL 7R+) population from murine fetal liver was investigated using a clonal assay capable of determining the potential of a progenitor to give rise to myeloid, T, and B cells. Unipotent progenitors generating T cells (p-T) or B cells (p-B) but not other types of progenitors were found in the IL-7R+ population. A large proportion of progenitors at day 12 of gestation are p-T, whereas the frequency of p-T dramatically decreases with gestational age. In marked contrast, p-B are very rare by day 12, but they rapidly increase thereafter. These findings strongly suggest that the commitment of multipotent progenitors to T and B cell lineages occurs independently. PMID- 10795744 TI - Rabeprazole versus omeprazole in preventing relapse of erosive or ulcerative gastroesophageal reflux disease: a double-blind, multicenter, European trial. The European Rabeprazole Study Group. AB - Gastroesophageal reflux disease (GERD) is a chronic condition, with 50-80% of patients experiencing recurrence within one year of completing initial treatment. In patients with erosive GERD, proton-pump inhibitors (PPI) provide faster healing and symptom relief than do H2-receptor antagonists and have become the treatment of choice. Rabeprazole is a new PPI with demonstrated efficacy in both the acute and maintenance treatment of erosive GERD. The primary objective was to compare efficacy and tolerability of rabeprazole and omeprazole in preventing relapse of healed erosive GERD. Secondary objectives included comparison of efficacy in preventing GERD relapse symptoms and in maintaining quality of life. In this multicenter, double-blind, parallel-group study, 243 patients with healed erosive GERD were randomised to receive rabeprazole 10 mg once daily in the morning (QAM) (N = 82); rabeprazole 20 mg QAM (N = 78); or omeprazole 20 mg QAM (N = 83). Endoscopies were performed at weeks 13, 26, 39 (if clinically indicated), and 52, or when symptoms suggested recurrence. Corpus biopsies were performed at each endoscopy, and antral biopsies were performed at study entry and exit. Rabeprazole 10 mg and 20 mg QAM were equivalent to omeprazole 20 mg QAM for all efficacy parameters. At week 52, relapse rates in the intent-to-treat populations were 5%, 4%, and 5% for rabeprazole 10 mg and 20 mg and omeprazole 20 mg, respectively. All treatments were well tolerated. In conclusion, both rabeprazole 10 mg and 20 mg QAM are equivalent to omeprazole 20 mg QAM in preventing recurrence of erosive GERD. PMID- 10795745 TI - Significance of autonomic nervous system activity in functional dyspepsia. AB - The purpose of this study was to examine if patients with functional dyspepsia could be separated into meaningful groups based on their autonomic function. Subjects were divided into two groups, and symptoms, gastric myoelectrical activity, gastric emptying, and psychological factors were compared. Group 1 had less autonomic variability but more cardiac reactivity than group 2. Symptom reports did not differ between groups. Group 1 had higher neuroticism scores than group 2, while group 2 showed greater tachyarrhythmia in response to drinking water than group 1. The relatively low autonomic variability in group 1 is consistent with higher sympathetic activity and may be associated with the group's greater neuroticism. The relative lack of cardiac reactivity in group 2 is consistent with lack of autonomic flexibility and may be related to the tachyarrhythmia observed in that group. The results of this study suggest that autonomic function may play a significant role in functional dyspepsia. PMID- 10795746 TI - Cyclin D1 expression is useful as a prognostic indicator for advanced esophageal carcinomas, but not for superficial tumors. AB - The purpose of the present study was to define the overexpression of cyclin D1 in superficial and advanced esophageal carcinomas and to investigate whether the expression of this molecule indicates a poor prognosis. This study included 41 patients with superficial esophageal carcinomas (Tis and T1) and 48 patients with advanced esophageal carcinomas (T2, T3, and T4). The expression of cyclin D1 in surgically resected specimens was evaluated immunohistochemically with a monoclonal antibody. Positive immunoreactivity was found in 31 of 89 cases (35%). Overexpression of cyclin D1 did not correlate with TNM classification or histologic type. Of the 48 patients with advanced esophageal carcinomas, 32 patients with cyclin D1-negative tumors survived longer than did 16 patients with cyclin D1-positive tumors (P = 0.0017). In contrast, we observed no survival difference between patients with cyclin D1-positive and -negative superficial esophageal carcinoma. These results suggest that cyclin D1 indicates a poor prognosis in cases of advanced esophageal carcinoma but not in cases of superficial esophageal carcinoma. PMID- 10795747 TI - Establishment of novel human esophageal cancer cell line in relation to telomere dynamics and telomerase activity. AB - The telomere and the telomerase in human esophageal cancer are not yet completely understood. The regulatory mechanism of telomerase activity and telomere dynamics has drawn considerable attention. It is generally assumed that when telomerase has been activated, no further telomere shortening should ensue; however, a much more complex pattern of telomere dynamics may exist in telomerase-positive cancer cells. A novel human esophageal cancer cell line (KAN-ES) was established and characterized. Using KAN-ES and its serially passaged subclones up to the 55th generation, we determined the alteration of telomere length (TRF), telomerase activity (TA), telomerase RNA expression (hTR), population doubling time, karyotype, and cytokeratin 14 expression during the process of establishing a cancer cell line. We found that the TRF was maintained between 4.0 and 5.0 kb during the serial passages, despite sustained high TA (assessed by an in vitro TRAP assay). No close relationships were found among TRF, TA, and hTR expression. TA and telomere dynamics were not associated with cellular growth ability and differentiation. However, the number of population doublings showed significant correlations with both the TA and doubling times. In conclusion, these dissociations between telomere dynamics and TA support the existence of additional controls on TRF in cancer cells. KAN-ES and its restored subclones should prove a valuable resource for esophageal cancer research. PMID- 10795748 TI - Neutrophil accumulation in development gastric ulcer induced by submucosal injection of endothelin-1 in rats. AB - Submucosal injection of endothelin (ET)-1 induces gastric ulcer. We investigated the roles of neutrophils and adhesion molecules (intercellular adhesion molecule (ICAM)-1 and CD18) in the development of ET-1-induced ulcers in rats. Ulcers were induced by submucosal injection of ET-1. Rats were injected with anti-neutrophil serum or F(ab')2 fragments of irrelevant mouse IgG2a (control), anti-ICAM-1 antibody, or anti-CD18 antibody. Ulcer tissues were subjected to measurement of myeloperoxidase (MPO) activity, ulcer size, and immunohistochemical study. Within 3 hr, arterial vasoconstriction and vascular congestion were observed at sites of ET-1 injection. By 6 hr, vascular congestion had disappeared, and ICAM-1 expression had markedly increased in venules in deep portions of the mucosa and submucosa, accompanied by an increase in the number of CD18-positive neutrophils. By 48 hr, ulcers that extended into the submucosa had developed. In controls, MPO activity gradually increased and was maximal by 6 hr. Neutrophil depletion, and immunoneutralizing of ICAM-1 and CD18 inhibited the increase in MPO activity, and decreased ulcer sizes measured at 48 hr. In conclusion, ET-1 causes ischemia reperfusion injury, and neutrophil accumulation after reperfusion mediated by the ICAM-1-CD18 pathway may be important in the development of ET-1-induced gastric ulcer. PMID- 10795749 TI - Pathogenesis of increased sucrose permeability in H. pylori gastritis. PMID- 10795750 TI - Obesity potentiates AOM-induced colon cancer. AB - Obesity and diet affect the incidence and severity of various types of cancer, including colon cancer. It is not known whether obesity, independent of diet, is a risk factor for colon adenocarcinoma. We used azoxymethane (AOM) to induce colon cancer in mature genetically obese male Zucker rats (fa/fa) on low-fat crude diet (LFC, 10% fat) and their lean counterparts (Fa/fa and Fa/fa) on high fat crude diet (HFC, 40% fat) for three months. At death visible tumors, histopathology, and colonic aberrant crypt (AC) formation were studied by blinded investigators. At death the obese animals were heavier (719 +/- 19 g; mean +/- SEM) than lean animals regardless of diet or genotype (Fa/fa-LFC:451 = 6 g; Fa/fa HFC:441 +/-10 g; Fa/Fa-HFC:412 +/- 9 g; P < 0.001 vs fa/fa by ANOVA). All AOM treated rats developed AC, compared to none of the saline-injected controls. Macroscopic adenocarcinoma developed in 8/9 obese rats on LFC (P < 0.001), compared to none in lean rats regardless of diet. Obese rats had significantly more AC (876 +/- 116) than any of the lean rats (Fa/fa-LFC:550 +/- 99; Fa/fa HFC:325 +/- 37; Fa/Fa-HFC:360 +/- 36; P < 0.05 vs fa/fa). We conclude that obesity more than exposure to high-fat diet was associated with colon carcinogenesis in these rats. PMID- 10795751 TI - Effect of cholesterol levels on villous histology in colonic adenomas. AB - To date no studies have examined the relationship between cholesterol levels and the occurrence of specific colonic polyp histologies. Villous histology has a greater predilection for subsequent malignancies than other histologies. Consequently, we examined the effect of cholesterol levels on the occurrence of villous adenomas. Just under one in 10 (9.5%, 15/158) patients had polyps with villous histologies. Cholesterol levels were positively and nonlinearly associated with a greater likelihood of villous histology, suggesting that a threshold exists for the effect of cholesterol level on the likelihood of having polyps with villous histology [odds ratios (OR) for combined two variable quadratic effect: cholesterol OR, 1.18; 95% confidence interval (CI), 1.02-1.37 and cholesterol squared OR, 1.004; 95% CI, 1.00-1.02]. Our data suggest that, in patients with polyps, higher cholesterol levels increase the likelihood of having polyps with villous histology, but that the effect of cholesterol level reaches a threshold. PMID- 10795752 TI - Carcinoma of the gallbladder--is it a sequel of typhoid? AB - Gallbladder diseases, including carcinoma, are common in the northern part of India and so are Salmonella typhi infection and typhoid carrier state. This study was aimed to find out the association of typhoid carrier state in patients with cholelithiasis, carcinoma of the gallbladder, and controls. The three groups are comparable in age and sex composition. This is the first study of its kind from an area of high endemicity for both typhoid infection and carcinoma of the gallbladder. A case-control study was carried out to detect typhoid carrier state among the patients with biliary diseases and healthy controls, using indirect haemagglutination assay measuring antibodies against highly purified S. typhi Vi polysaccharide antigen. A significantly high Vi positivity was observed in patients with gallbladder carcinoma (29.4%) compared to controls (5%) (chi2 = 6.325, P < 0.004, OR = 7.19) and patients with cholelithiasis (10.7%) (chi2 = 5.066, P < 0.01, OR = 3.86). There is 8.47 times more risk of developing carcinoma of the gallbladder in culture-positive typhoid carriers than the noncarriers. The present study suggests the typhoid carrier state to be one of the possible mechanisms of gallbladder carcinogenesis. PMID- 10795753 TI - Stimulation of gallbladder by intravenous infusion of amino acid: a new method to obtain duodenal bile for bile analyses. AB - Bile analysis is performed on duodenal bile obtained after stimulation of the gallbladder (GB) with cholecystokinin or its analog. Rapid amino acids (AA) infusion contracts the GB. Our objective was to assess qualitatively the adequacy of duodenal bile obtained following AA infusion. We prospectively studied 15 patients each with non ulcer dyspepsia (NUD) and gallstones (GS), and 17 patients with acute idiopathic pancreatitis (AIP). Duodenal bile was obtained after duodenal intubation and stimulation of GB by rapid AA infusion. The GB contractile response to AA infusion was assessed by functional ultrasonography. Bile was analyzed for lipids, nucleation time, cholesterol saturation index (CSI), and biliary crystals. Adequate duodenal bile could be collected in all but one patient with AIP and one with GS. The mean +/- SD percent ejection fraction of GB following AA infusion in patients with NUD, GS and AIP was 53.41 +/- 16.23, 52.06 +/- 16.07, and 43.37 +/- 14.16, respectively. Biliary microscopy showed the presence of cholesterol monohydrate crystals (CMC) in 7 of 16 patients with AIP, 12 of 14 patients with GS, and 1 of 15 patients with NUD. Mean CSIs in patients with NUD, GS, CMC-positive AIP, and CMC-negative AIP were 0.78, 1.97, 1.85, and 1.00, respectively. Nucleation time in patients with NUD and gallstones was 21.1 and 8.42 days. respectively. In conclusion, qualitatively adequate bile can be obtained for chemical and microscopic examination following rapid intravenous infusion of AA to stimulate the GB. PMID- 10795754 TI - Effect of peritoneal lavage and lymph ligature on systemic complications of experimental acute pancreatitis. AB - We studied the involvement of ascitic fluid on the systemic effects of experimental acute pancreatitis. This has been achieved by comparing the effectiveness of either peritoneal lavage or lymphatic ligature on preventing changes in systemic vascular permeability. Three hours after induction of pancreatitis, we found increases in vascular permeability in the pancreas, lung, and intestine. Both peritoneal lavage and lymphatic ligature were able to prevent the changes observed in the lung and intestine and the increases on plasma levels of lipase and amylase, suggesting a similar involvement for lymphatic draining and peritoneal absorption pathways. In addition, we evaluated the effect of intraperitoneal deposition into health rats of pancreatitis-associated ascitic fluid collected from rats with experimental acute pancreatitis. A significant increase in plasma amylase and lipase levels could be observed but no changes in vascular permeability were found. Altogether, these results indicate that transperitoneal absorption of toxic mediators from the ascitic fluid is not enough to explain the systemic damage induced by acute pancreatitis. PMID- 10795755 TI - Lipopolysaccharide directly affects pancreatic acinar cells: implications on acute pancreatitis pathophysiology. AB - We have explored whether lipopolysaccharide (LPS, endotoxin) induces pancreatic injury on pancreatic acinar cells both in vivo and in vitro. Wistar male rats were treated with four intraperitoneal injections of 10 mg/kg LPS, and AR4-2J cells were exposed to increasing doses of LPS. Expression of pancreatitis associated-protein (PAP) mRNA was strongly induced in AR4-2J cells exposed to LPS, while amylase mRNA was reduced. LPS also induced apoptosis and expression of TNF-alpha, IL-1beta, and IL-8 mRNA in AR4-2J cells. The in vivo effect of LPS showed structural signs of cellular damage, including numerous cytoplasmic vacuoles, severe nuclear alterations, and high expression of PAP mRNA. This study demonstrated that LPS induced pancreatic damage by directly affecting the pancreatic acinar cells. The role of LPS in the pathophysiology of acute pancreatitis may be partly due to the effect LPS has on the acinar cell. PMID- 10795756 TI - Cytokine production by CAPAN-1 and CAPAN-2 cell lines. AB - Recently, there has been a great deal of interest in the role of cytokines in acute pancreatitis. Serum levels of IL-1, IL-6, and TNF-alpha have been demonstrated to be elevated in acute pancreatitis. We hypothesized that cytokines may be produced primarily by pancreatic parenchymal cells. Reasoning that ductal epithelium is the cell type most likely to be exposed to noxious stimuli in common causes of pancreatitis, such as ERCP and passage of a gallstone, we examined the response of well differentiated pancreatic ductal adenocarcinoma cell lines to stimuli known to stimulate cytokine production in other cells. CAPAN-1 and CAPAN-2 cells were incubated with endotoxin or TNF-alpha. The supernatant was assayed for production of IL-1, IL-6, and IL-8 by ELISA. The cells were assayed for activation of the transcription factor NF-kappaB by electrophoretic mobility shift assay. There was no detectable production of IL-1 by either cell line. CAPAN-1 cells had concentration-dependent production of IL-6 and IL-8 in response to both endotoxin and TNF-alpha. CAPAN-2 cells had concentration-dependent production of IL-6 and IL-8 in response to TNF-alpha. They had low level expression of IL-8 that was unaffected by any concentration of LPS, and no detectable production of IL-6 in response to LPS. These findings suggest that pancreatic duct cells may take an active part in the pathogenesis of acute pancreatitis through the production of cytokines. PMID- 10795757 TI - Increased serum leptin by cholecystitis in a diabetic patient. PMID- 10795758 TI - Erythromycin enhances solid-phase gastric emptying in induced-hyperglycemia in patients with truncal vagotomy and pyloroplasty. AB - Erythromycin has been found to be a gastrointestinal prokinetic agent while acute hyperglycemia has been associated with delayed gastric emptying in healthy controls and diabetics. The aim of this study was to investigate whether hyperglycemia, per se, alters gastric motility, during erythromycin-induced acceleration of gastric emptying of solids in patients with truncal vagotomy and pyloroplasty (TVP) and the role of vagus nerves. Eight TVP patients and six controls underwent scintigraphic measurement of gastric emptying of a solid meal, during placebo in normoglycemia (5-8.9 mmol/liter glucose) or 200 mg intravenous erythromycin lactobionate in normo- or hyperglycemia (16-19 mmol/liter glucose) induced by intravenous glucose infusion, on separate days in random order. In the TVP patients during normoglycemia, the erythromycin compared to placebo accelerated the meal gastric half-emptying time (T1/2), (37.12 +/- 6.87 vs 91.88 +/- 11.53, P < 0.001) and decreased the lag-phase duration (P < 0.001) and the percentage of meal retained in the stomach at 120 min (P < 0.001). Erythromycin in hyperglycemia compared to normoglycemia increased T1/2 (61.25 +/- 10.67 vs 37.12 +/- 6.87, P < 0.001), prolonged lag-phase duration (P < 0.001), and the percentage of isotope retained in the stomach at 120 min (P < 0.001). The T1/2, the lag phase duration, and the meal retained in the stomach at 120 min, after giving placebo was significantly increased, compared to erythromycin administration in hyperglycemia (P < 0.001). Significant differences among patients and controls were found during gastric emptying after giving placebo and after erythromycin in hyperglycemia (P = 0.04 and P = 0.007, respectively), while nonsignificant differences were found after giving erythromycin in normoglycemia. We conclude that the effect of erythromycin-induced acceleration on gastric emptying is related to the plasma glucose level. Hyperglycemia reduces the erythromycin-induced acceleration of gastric emptying of solids in both controls and TVP patients. A significant increase in the delay of gastric emptying was achieved in TVP patients compared to controls after giving erythromycin in hyperglycemia and after placebo. Despite the inhibitory effect of induced hyperglycemia on gastric emptying, erythromycin is still able to accelerate the emptying rate and could prove to be a useful prokinetic agent under hyperglycemic conditions. Hyperglycemia may indicate a cholinergic-antagonist pathway that delays the erythromycin-induced acceleration of gastric emptying of solids and is more evident in vagotomized patients than controls, who retain the functional integrity of the vagus nerves. PMID- 10795759 TI - Cholera toxin-induced secretion in rats is reduced by a soluble fiber, gum arabic. AB - Gum arabic (GA), a soluble fiber with emulsifying properties, enhances intestinal water and electrolyte absorption in normal and secreting rats. Our aim was to assess the effect of GA, 2.5 and 5.0 g/liter, on cholera toxin-induced water and electrolyte secretion in rat jejunum in vivo. After a 2-hr exposure to cholera toxin, jejunal segments of adult rats were perfused in vivo with at plasma electrolyte solution containing GA, 0, 2.5 or 5.0 g/liter. 24Na was used as a marker of sodium influx. Cholera toxin-induced secretion was reduced by GA, 2.5 and 5.0 g/liter. 24Na secretion into the lumen was reduced by GA. GA caused a morphological expansion of intercellular spaces in the villi but not crypts. In conclusion, GA promotes lumen to blood intestinal transport of water and sodium despite cholera toxin activation. These observations support a potential role for GA in enhancing the efficacy of ORS. PMID- 10795760 TI - Sleep disturbance influences gastrointestinal symptoms in women with irritable bowel syndrome. AB - This analysis evaluated the association between sleep disturbance and gastrointestinal symptoms in women with and without irritable bowel syndrome (IBS), and examined the role of psychological distress in this relationship. Women with IBS (N = 82) reported considerably higher levels of sleep disturbance compared to controls (N = 35), using both retrospective seven-day recall and daily diary recall for two menstrual cycles (P < 0.05 on 8 of 10 measures). We used daily diary data to estimate the association between sleep disturbance and gastrointestinal symptoms, both across women (ie, whether women with high average sleep disturbance have higher average gastrointestinal symptoms) and within woman (ie, whether poorer than average sleep on one night is associated with higher than average gastrointestinal symptoms the following day). The regression coefficients for the across-women effect are large and highly significant in both groups (IBS, beta +/- SE = 0.46 +/- 0.08, P < 0.001; controls, 0.57 +/- 0.13, P < 0.001). The regression coefficients for the within-woman effect are considerably smaller and statistically significant only in the IBS group (IBS, 0.06 +/- 0.02, P = 0.006; control, 0.01 +/- 0.03, P = 0.691). These regression coefficients showed little change when daily psychological distress or stress was controlled for, the one exception being the coefficient for the across-women effect in the IBS group, which decreased substantially but still remained highly significant. Because it is possible that gastrointestinal symptoms could, in fact, cause poor sleep, we also fitted the temporally reversed model to evaluate the association between gastrointestinal symptoms on one day and sleep disturbance that night. The within-woman regression coefficients were nonsignificant in both the IBS and control groups. In conclusion, these results are consistent with the hypothesis that poor sleep leads to higher gastrointestinal symptoms on the following day among women with IBS. PMID- 10795761 TI - Effects of several endothelin receptor antagonists on gastrointestinal transit of guinea pigs. AB - This study was undertaken to investigate the effects of endothelin(A) receptor antagonist (ET(A)-RA) BQ485; ET(B)-RA BQ788, and nonselective ET(A/B)-RA Bosentan on the gastrointestinal transit of guinea pigs. We further analyzed the distribution of ET-R subtypes on smooth muscle cells (SMC) of the gastrointestinal tract to investigate their direct involvement on SMC in gastrointestinal tract transit. A guinea pig model was used to measure intestinal transit. The effects of Bosentan (100 mg/kg, per os), BQ485 (1 mg/kg, intraperitoneally) and BQ788 (1 mg/kg, intraperitoneally) on transit in stomach, small intestine, and colon were evaluated. We separated SMC from stomach, small intestine, cecum, and colon by collagenase and analyzed the distribution of ET-R subtypes in each part by binding assay. Gastric transit and colon transit were significantly inhibited by BQ485, BQ788, and Bosentan. Small intestinal transit was not affected by any of these agents. ET(A)-R and ET(B)-R were widely distributed on SMC of stomach, small intestine, cecum, and colon. The ratio of ET(A)-R to ET(B)-R was 1:3 in stomach, small intestine, and cecum, but was 1:10 in colon. The ratio of the total number of ET-R on SMC of stomach, small intestine, cecum, and colon was 1:3:9:1. These results indicate that both ET(A)-R and ET(B)-R are strongly involved in the transit in the stomach and colon. However, the discrepancy between the effects of the various ET-R antagonists on gastrointestinal transit and the distribution of ET-R on SMC of the gastrointestinal tract suggests that ET-R on SMC of the gastrointestinal tract is not directly involved in gastrointestinal transit. PMID- 10795762 TI - Effects of free radicals and leukocytes on increases in blood-brain barrier permeability during colitis. AB - This study examined the role of leukocytes and free radicals on increases in permeability of the blood-brain barrier (BBB) in rabbits with acute colitis. Trinitrobenzene sulfonic acid (TNBS) was used to induce colitis in male New Zealand White rabbits. The extraction ratio of fluorescein was used as an index of the permeability of the BBB. The extraction ratio for fluorescein was 1.0 x 10(-3) +/- 2 x 10(-4) ml/g and 1.1 x 10(-3) +/- 3 x 10(-4)ml/g (mean +/- SE) for saline (N = 7) and ascorbic acid-treated (N = 8) rabbits with sham colitis. Conversely, TNBS-induced acute colitis increased the extraction ratio over 70% in saline (N = 8) and ascorbic acid-treated (N = 8) animals. Vinblastine significantly reduced the number of circulating leukocytes, whereas the permeability of the BBB was augmented by 54% in rabbits with TNBS-induced acute colitis (N = 8). Vinblastine had no effect on the permeability of the BBB in rabbits with sham colitis (N = 8). These data suggest that free radicals are not responsible for BBB disruption, and leukocytes may protect the BBB during acute colitis. PMID- 10795764 TI - Prediction of short-term outcome for patients with active ulcerative colitis. AB - We report results of a retrospective chart review to evaluate factors predicting short-term outcome of patients with ulcerative colitis treated by corticosteroids. Between January 1992 and December 1997, we treated 71 patients with ulcerative colitis (44 with severe and 27 with moderately severe disease). Forty-nine patients were treated by conventional prednisolone therapy and 22 patients by steroid pulse therapy. There were no differences in clinical or endoscopic improvement between the two treatments. Clinical examination showed that 41 patients entered remission, 17 patients improved, and 13 patients did not respond. Endoscopically, 26 patients entered remission, 30 patients improved, and 15 patients did not respond. Extent of disease, type of disease (first attack, relapsing, or chronic active type), and endoscopic findings were factors useful in predicting short-term outcome of medical treatment. PMID- 10795763 TI - Combined oral sodium butyrate and mesalazine treatment compared to oral mesalazine alone in ulcerative colitis: randomized, double-blind, placebo controlled pilot study. AB - Butyrate represents the main source of energy for colonic epithelial cells; however, its availability/utilization is impaired in ulcerative colitis (UC). In the present randomized, double-blind, placebo-controlled pilot study, the safety and efficacy of colonic targeted oral sodium butyrate tablets, coated with a pH dependent soluble polymer, have been evaluated in ulcerative colitis. Thirty patients with mild to moderate colitis underwent a six-week course of oral sodium butyrate (4 g/day) plus oral mesalazine (2.4 g/day), (Group A) or of oral mesalazine plus placebo (Group B). Clinical, endoscopic, and histologic data were collected at the beginning and the end of the study. Twenty-five patients completed the study (12 in group A, 13 in group B). No untoward side effects were reported. In group A, seven patients underwent remission and four improved; in Group B the numbers were 5 and 5, respectively. After treatment, all clinical parameters had significantly improved in both treatment arms compared to pretreatment findings. The UC disease activity index (UCDAI) score decreased from 7.27 +/- 2.02 to 2.58 +/- 2.19 (P < 0.05) in the combined treatment group and from 6.07 +/-1.60 to 3.46 +/- 1.98 (P < 0.05) in group B. The endoscopic and histologic scores also significantly improved after treatment in both groups (P < 0.05). The difference between the two treatment arms was not significant, but a significantly better improvement vs baseline values (P < 0.05) was observed in the combined treatment group vs the mesalazine group, when considering both the clinical index (delta9.58 +/- 4.19 vs 5.92 +/- 3.48) and the UCDAI score (delta4.67 +/- 2.19 vs 2.54 +/- 2.18). A more favorable trend, although not significant, was observed for all individual parameters in group A. In conclusion, results of the present pilot study indicate that oral butyrate is safe and well tolerated. These data also suggest that oral butyrate may improve the efficacy of oral mesalazine in active ulcerative colitis and prompt the need of a large scale investigation to confirm the present findings. PMID- 10795766 TI - Persistent Clostridium difficile colitis: a possible etiology for the development of collagenous colitis. PMID- 10795765 TI - Recombinant factor XIII improves established experimental colitis in rats. AB - Factor XIII (FXIII) is the plasma-borne transglutaminase involved in fibrin clot stabilization and wound healing. FXIII levels in the plasma of patients with inflammatory bowel diseases are lower than normal and there is a significant inverse correlation of FXIII levels with clinical severity. Moreover, uncontrolled studies report beneficial effects of FXIII supplementation in patients resistant to conventional therapies. We investigated the effects of intravenous recombinant FXIII (rFXIII) treatment in experimentally induced rat colitis to verify that FXIII was the active agent in plasma FXIII concentrates and to better understand the potential therapeutic use of this protein. Colitis was induced by instillation of 12% 2.4,6-trinitrobenzenesulfonic acid (TNBS) in 50% ethanol into the colon of male Wistar rats. Rats were treated with 0.65 mg/kg rFXIII or vehicle (intravenously) daily for 10 days. Treatment was started either immediately after TNBS/EtOH instillation (to evaluate effects on developing lesions) or seven days later (to evaluate effects on established lesions). In both cases rats were killed either immediately after the end of treatment (to evaluate immediate effects) or 17 days later (to evaluate long-lasting effects). The effects of rFXIII were compared to positive (5-amino-2-hydroxybenzoic acid) control over a 35-day time course. The severity of lesions was determined by colon weight and macroscopic and histologic scores. Transglutaminase activity was measured in both colon tissue and serum. rFXIII treatment reduced lesion severity significantly not only in developing but also in established lesions. Improvements in healing persisted at least 18 days after treatment was discontinued. Serum and tissue transglutaminase levels were restored by rFXIII treatment. In conclusion, pure rFXIII is as effective as plasma FXIII concentrates in a rat model of experimental colitis. In addition, rFXIII significantly improves the healing of preexisting lesions, a characteristic useful in treatment of human inflammatory bowel diseases. PMID- 10795767 TI - Estimation of nitrate metabolism in intestinal tract by measuring breath nitrous oxide concentration in Chinese and Japanese. AB - We have previously reported that ingestion of vegetables containing high nitrate (NO3-) increases breath nitrous oxide (N2O) concentration, probably due to denitrification. In the present study, we estimated NO3- metabolism in the intestine by determining exhaled breath N2O concentration after the ingestion of vegetables by 16 healthy Chinese and Japanese. Breath samples were collected at 15-min intervals for 4 hr after subjects ingested 180 g of vegetable juice or 50 g of lettuce. Breath N2O concentration was measured by an IR-PAS analyzer. Lettuce but not vegetable juice increased N2O concentrations similarly in Japanese and Chinese. In control subjects who ingested nothing, there were significant differences between Chinese and Japanese in peak N2O concentrations (334 +/- 91 vs 140 +/- 24 ppb, P = 0.027) and total excretions (535 +/- 143 vs 214 +/- 36 microg, P = 0.036). Although the reason for this difference is unclear, Chinese subjects could produce breath N2O from other metabolic pathways than denitrification of dietary NO3. PMID- 10795768 TI - Improvement of chronic diarrhea after treatment for intestinal spirochetosis. PMID- 10795769 TI - Oral or intravenous lidocaine administration to perform megx test? PMID- 10795770 TI - Endoscopic, ultrasonographic, and US-Doppler parameters as indicators of variceal bleeding in patients with schistosomiasis. AB - Bleeding from esophagogastric varices is a potentially deadly complication in patients with hepatosplenic schistosomiasis. The aim of this study is to establish indicators of variceal bleeding. We studied 40 patients with compensated hepatosplenic schistosomiasis and varices, analyzing four endoscopic (variceal size, red color signs, fundic varices, and congestive gastropathy), nine ultrasonographic (right and left hepatic lobe size, periportal and gallbladder wall thickness, portal and splenic veins diameter, spleen longitudinal axis and volume, and presence of collateral circulation), and five US-Doppler parameters (portal and splenic veins velocity and flow and portal vein congestion index). Patients were divided in two groups according to previous history of variceal bleeding. The group with bleeding episodes was again divided in two groups: with and without treatment, namely endoscopic sclerotherapy. All endoscopic parameters and two ultrasonographic (periportal thickness and portal vein diameter) were statistically different between the groups with and without previous bleeding. The likelihood index, adopted to determine the best parameters related to previous bleeding showed that the most important combinations are: gastropathy and red signs followed by portal vein diameter and variceal size. In conclusion, although aware of the limits of the statistical analysis due to the small number of patients, our results demonstrated that endoscopic and US parameters (isolated or combined) can identify patients with a high risk of variceal bleeding, allowing physicians to optimize prophylactic therapy. PMID- 10795771 TI - 48-hour hemodynamic effects of octreotide on postprandial splanchnic hyperemia in patients with liver cirrhosis and portal hypertension: double-blind, placebo controlled study. AB - Octreotide is effective during 48 h in the treatment of acute variceal bleeding, probably by reducing variceal blood flow and pressure. Its basal and postprandial effects on splanchnic and systemic hemodynamics, and hormonal changes over this time interval have not yet been studied. Twenty-four patients with cirrhosis and portal hypertension were randomized to receive a liquid meal and either octreotide (Oct, 100 microg bolus intravenous, followed after 2 h by a continuous infusion of 25 microg/hr for 20 hr) or placebo (Plac) given at three consecutive days. Splanchnic (Doppler ultrasound) and systemic hemodynamics (noninvasive cardiac monitoring) were assessed on four consecutive days (one control day and three treatment days) during 2 hr. The postprandial increase in mean blood velocity of the superior mesenteric artery (SMA-V(mean) +44%), portal blood velocity (PV-V(mean), +44%) and total hepatic blood flow (HBF, +40%) observed in the placebo group during the control day was abolished during the first day of treatment (SMA-V(mean), +3%, P < 0.01; PV-V(mean), +6%, P < 0.05; HBF, -25%, P < 0.01) and still reduced after 48 hr in the octreotide group (SMA-V(mean) +28%, P < 0.05; PV-V(mean), +22%, P > 0.05; HBF, -8%, P < 0.05). The postprandial increase in cardiac index (CI, + 10%) and decrease in systemic vascular resistance index (SVRI, -6%) were blunted after the initial injection of octreotide only (CI, -8%, P < 0.05; SVRI, +18%, P < 0.01). Endothelin-1-levels, which were increased at baseline (Plac 25 +/- 17, Oct 16 +/- 13 ng/liter, P > 0.05) decreased significantly after 48 hr of treatment with octreotide (Plac 27 +/- 20, Oct 8 +/- 4 ng/liter, P < 0.05). Octreotide is effective during 48 hr in the prevention of postprandial hyperemia in cirrhotics, even if its efficacy is decreasing over time. Moreover it may have positive effects on systemic vasodilation in cirrhotics. These findings suggest a potential role of this drug in the chronic treatment of portal hypertension. PMID- 10795772 TI - Nature and behavior of serum cytokines in type 1 autoimmune hepatitis. AB - To assess the relationship between serum cytokine behavior and treatment outcome in type 1 autoimmune hepatitis, serum levels of interferon-gamma, interleukin-2, interleukin-4, and interleukin-10 were measured by enzyme immunoassay in 43 patients and 20 normal subjects. Serum samples were similarly tested in 38 patients after corticosteroid treatment. Serum levels of interleukin-2 and interleukin-4 were significantly lower in patients than in normal subjects. Interleukin-2 was the least common cytokine detected before (3%), during (0%), or after treatment (0%). Serum levels of interleukin-10 at presentation did not differ from those of normal subjects but they did decrease during therapy, especially in patients who entered remission. Changes in these levels, however, did not always parallel treatment outcome or histological activity. We conclude that serum levels of interleukin-2 and interleukin-4 are lower than normal in type 1 autoimmune hepatitis. Serum concentrations of interleukin-10 diminish during corticosteroid therapy but changes do not closely reflect outcome. The rarity of interleukin-2 in serum may be a distinguishing feature. PMID- 10795773 TI - Reversible nefazodone-induced liver failure. PMID- 10795774 TI - Severe but reversible hepatitis induced by troglitazone. PMID- 10795775 TI - The relationship between %HRpeak and %VO2peak in patients with chronic obstructive pulmonary disease. AB - PURPOSE: The relationship between the percent peak oxygen consumption (%VO2peak) and percent peak heart rate (%HRpeak) has been suggested as a method for exercise prescription. According to the American College of Sports Medicine (ACSM), 50, 60, 80, and 85% of VO2peak represent 62, 70, 85, and 90% of HRpeak, respectively. Recent research with young healthy adults has shown that this relationship is not as ACSM suggests. Furthermore, it has been suggested that this relationship be examined in populations with low functional capacity. METHODS: The present study evaluated the relationship between %HRpeak and %VO2peak in 125 patients with chronic obstructive pulmonary disease before and then after 3 months of exercise intervention. RESULTS: The mean percentages of HRpeak for the COPD patients at baseline were 70, 76, 87, and 90% at 50, 60, 80, and 85% of VO2peak. Percentages of HRpeak at 50, 60, and 80% of VO2peak were significantly different from those suggested by ACSM. The mean percentages of HRpeak after the 3-month exercise intervention were 71, 77, 87, and 90% at 50, 60, 80, and 85% of VO2peak. Percentages of HRpeak at 50, 60, and 80% of VO2peak were significantly different from those suggested by ACSM. There were no significant differences between the slopes and the intercepts of the two equations used to describe the relationship between %HRpeak and %VO2peak. Therefore, we pooled the data from the pre- and post-intervention tests to form the following equation to describe the relationship between %HRpeak and %VO2peak for COPD patients: %HRpeak = 0.55 (+/- 0.01)%VO2peak + 43.2 (+/- 1.4). CONCLUSION: In conclusion, the relationship between %HRpeak and %VO2peak described by ACSM is not applicable to patients with COPD. Furthermore, this relationship does not change with exercise training and can be described by a single equation. PMID- 10795776 TI - Cost effectiveness analysis of screening of high school athletes for risk of sudden cardiac death. AB - Sudden cardiac death of a high school athlete is an alarming tragedy. Three preparticipation screening methods have been recommended to reduce its occurrence: specific cardiovascular history and physical examination, 12-lead ECG, and two-dimensional (2D) echocardiography. This study analyzes the cost effectiveness of each of these methods. The cost to perform each test and to evaluate abnormal screening findings were approximated. The years of life gained through detection of athletes with potential causes of sudden cardiac death were estimated. Overall, the approximate costs per year of life saved for the preparticipation cardiovascular screening examinations are: specific cardiovascular history and physical examination, $84,000; 12-lead ECG, $44,000; and 2D echocardiography, $200,000. The 12-lead ECG is the most cost effective preparticipation cardiovascular modality of the three currently recommended methods. Similar cost effectiveness for history and physical examination or 2D echocardiography would require respectively a 2-fold increase in sensitivity or 4.5-fold decrease in cost. PMID- 10795777 TI - The importance of gender on myokinetic deficits before and after microinjury. AB - PURPOSE: The purpose of this study was to determine the importance of gender on myokinetic deficits before and after muscle microinjury. METHODS: A repeated measures design assessed selected muscle force production characteristics in 20 male and 25 female volunteers. Peak force production (PFP) and the peak rate of force production (PRFP) were assessed before and over a 4-d period after an induced muscle microinjury. RESULTS: ANOVA revealed statistically significant mean (+/-SD) differences between men and women for PFP and PRFP (P < 0.0001). Both genders demonstrated significant between-day differences for PFP (P < 0.016), whereas only men demonstrated significant between-day differences for PRFP (P < 0.016). CONCLUSIONS: Our results reveal that muscle force generating capabilities of physically active men exceed that of women both before and after microinjury. Myokinetic deficits were most pronounced acutely, between 24 and 48 h postinjury, followed by a near complete recovery at day 4 (96 h postinjury). Both genders suffered acute and residual deficits for PFP, whereas only men showed significant acute deficits for PRFP. We recommend that athletes, both male and female, refrain from strenuous exercise at least 48 h postinjury, or until force-generating capabilities normalize. More research needs to be conducted to substantiate these findings. PMID- 10795778 TI - The effects of 17 alpha-methyltestosterone on myocardial function in vitro. AB - Testosterone analogs have been used as performance enhancers by athletes for more than 40 yr. We asked whether the anabolic steroid 17 alpha-methyl-4-androstene-17 ol-3-one (17 alpha-MT) would affect intrinsic contractile function of the heart. Male Sprague-Dawley rats, 125-150 g, were treated with 17 alpha-MT either parenterally or orally for up to 8 wk. Intrinsic contractile function of the hearts was assessed utilizing both the isolated working heart and isovolumic perfused heart preparations. Isolated working hearts from 17 alpha-MT-treated rats had a 45% decrease in heart work attributable largely to a similarly decreased stroke volume. Isovolumic perfused hearts from treated animals had elevated left ventricular systolic and diastolic pressures at similar interventricular volumes compared to controls. Rates of ventricular pressure development (+dP/dT) or relaxation (-dP/dT) were unchanged as a result of the treatment. However, static elastance was reduced in potassium-arrested hearts from the 17 alpha-MT treatment (63% increase in interventricular pressure), consistent with a limitation being imposed on stroke volume by a decreased myocardial compliance. Hydroxyproline content of the hearts was not altered by 17 alpha-MT treatment suggesting that increased stiffness was not a consequence of collagen proliferation. Treatment of the steroid rats with beta aminopropionitrile, a compound that inhibits lysyl oxidase, restored the left ventricular volume-pressure relationship (elastance curve) to that of control hearts. Thus, chronic treatment with anabolic steroids appears to reduce left ventricular compliance, possibly related to an enhanced activity of lysyl oxidase, and results in increased crosslink formation between collagen strands in the extracellular matrix. PMID- 10795779 TI - Fluid snacks to help persons with type 1 diabetes avoid late onset postexercise hypoglycemia. AB - PURPOSE: The present study assessed whether whole milk, skim milk, or two commercially available sports drinks are effective in preventing late onset postexercise hypoglycemia (LOPEH) in persons with type 1 diabetes mellitus. METHODS: Subjects ingested water, whole milk, skim milk, sport drink A (carbohydrate and electrolytes), or sport drink B (carbohydrate, fat, and protein) before, during, and after 1 h of bicycle exercise at 60% VO2max in the late afternoon. Drinks were isocaloric (470 +/- 150 kcal) and the number of calories consumed was based on individual energy expenditure. No adjustment in insulinization was allowed in anticipation of exercise. RESULTS: During water trials all subjects became hypoglycemic. Most drinks lead to a moderate hyperglycemia (range of mean values = 200-280 mg x dL(-1)) during the period between the end of exercise and dinner, but this was not the case for whole milk (range 80-120 mg x dL(-1)). Glycemia peaked about 1.5 h after dinner and declined over the next 90 min. Persistent hyperglycemia (range of means = 200-310 mg x dL( 1)) from after exercise to about 4 h postexercise was observed with sports drink B. A decline in glycemia in the evening was greatest during the skim milk trial and required subjects to ingest more carbohydrate as a late evening snack. The least decline during this period occurred during the whole milk trial. Subjects experienced pre-bed and early morning (0300 h) hypoglycemia in 7 of the 28 trials. CONCLUSIONS: These data show that whole milk and sports drinks that are designed for both quick (sport drink A) and long lasting (sport drink B) nutrient replenishment can be used by persons with type 1 diabetes in an effort to avoid LOPEH. PMID- 10795780 TI - Muscle, bone, and the Utah paradigm: a 1999 overview. AB - The still-evolving Utah paradigm of skeletal physiology supplements former ideas about the control of postnatal bone strength and "mass" in health and disease, on earth and in orbit, and as functions of physical activities including sports. In its view, peak muscle forces on bone dominate control of the biologic mechanisms that control changes in our postnatal whole-bone strength and mass. This contrasts with former ideas that chiefly nonmechanical factors dominate that control and mechanical factors have little influence on it. The newer view begs the question of how such a bone-muscle relationship would work, and the Utah paradigm offers plausible explanations for that. Whereas many biomechanicians, orthopedists, and sports medicine experts might find the newer view sensible, some people in other disciplines may not. This article summarizes some evidence that support the Utah paradigm and the above-stated view and explores some of the paradigm's features and implications. PMID- 10795781 TI - Blood hemostasis in exercise and training. AB - Formation of the blood clot is a slow but normal physiological process occurring as a result of the activation of blood coagulation pathways. Nature's guard against unwanted blood clots is the fibrinolytic enzyme system. In healthy people, there is a delicate dynamic balance between blood clot formation and blood clot dissolution. Available evidence suggests that exercise and physical training evoke multiple effects on blood hemostasis in normal healthy subjects and in patients. A single bout of exercise is usually associated with a transient increase in blood coagulation as evidenced by a shortening of activated partial thromboplastin time (APTT) and increased Factor VIII (FVIII). The rise in FVIII is intensity dependent and continues into recovery. The effects of acute exercise on plasma fibrinogen have yielded conflicting results. Thus, the issue of whether exercise-induced blood hypercoagulability in vitro mirrors an in vivo thrombin generation and fibrin formation remains disputable. Exercise-induced enhancement of fibrinolysis has been repeatedly demonstrated using a wide range of exercise protocols incorporating various exercise intensities and durations. Moderate exercise appears to enhance blood fibrinolytic activity without a concomitant activation of blood coagulation mechanisms, whereas, very heavy exercise induces simultaneous activation of blood fibrinolysis and coagulation. The increase in fibrinolysis is due to a rise in tissue-type plasminogen activator (tPA) and decrease in plasminogen activator inhibitor (PAI). The mechanism of exercise induced hyperfibrinolysis is poorly understood, and the physiological utility of such activation remains unresolved. Strenuous exercise elicits a transient increase in platelet count, but there are conflicting results concerning the effect of exercise on platelet aggregation and activation. Few comprehensive studies exist concerning the influence of exercise training on blood hemostasis, making future investigation necessary to identify whether there are favorable effects of exercise training on blood coagulation, fibrinolysis, and platelet functions. PMID- 10795782 TI - Evidence for an inadequate hyperventilation inducing arterial hypoxemia at submaximal exercise in all highly trained endurance athletes. AB - PURPOSE: The majority of highly trained endurance athletes with a maximal oxygen uptake greater than 60 mL x min(-1) x kg(-1) develop exercise-induced hypoxemia (EIH). Yet some of them apparently do not. The pathophysiology of EIH seems to be multifactorial, and one explanatory hypothesis is a relative hypoventilation. Nevertheless, conflicting results have been reported concerning its contribution to EIH. The aim of this study was to compare the cardiorespiratory responses to maximal exercise of highly trained endurance athletes demonstrating the same aerobic capacity without EIH (N athletes) and with EIH (H athletes). METHODS: Ten N athletes and twelve H athletes performed an incremental exercise test. Measurements of arterial blood gases and cardiorespiratory parameters were performed at rest and during exercise. RESULTS: All athletes presented a significant decrease in PaO2 (P < 0.05) from rest up to 80% VO2max associated with an increase in PaCO2, both findings consistent with a relative hypoventilation. Then the H athletes, who had a greater training volume per week and a higher second ventilatory threshold than the N athletes (respectively, 17 +/- 1.1 vs 13.1 +/- 0.7 h x wk(-1); 91.8 +/- 1.7 vs 86.1 +/- 1.8% VO2max), presented a continuous PaO2 decrease up to VO2max. This was associated with a widening (Ai-a)DO2. CONCLUSION: This study showed that a relative hypoventilation, probably induced by a high level of endurance training, induced hypoxemia in all athletes. However, a nonventilatory mechanism, perhaps related to the volume of training, seemed to affect gas exchanges beyond the second ventilatory threshold in the H athletes, thereby enhancing EIH. PMID- 10795783 TI - Exercise shifts the platelet aggregation modulatory role from native to mildly oxidized low-density lipoprotein. AB - PURPOSE: The role of low-density lipoprotein (LDL) lipid peroxides in strenuous exercise-induced changes in platelet function was studied in 30 patients (male/female = 22/8) aged 30-62 yr (mean +/- SD = 508). METHODS: All subjects were subjected to a treadmill exercise test, using the standard Bruce protocol. Blood samples were collected pre-, peak, and 10 min postexercise to assess hematological and biochemical parameters and platelet aggregation. Ex vivo whole blood platelet aggregation during treadmill exercise was assessed in 10 subjects by adding mildly oxidized LDL. RESULTS: Preexercise, a correlation existed between plasma thromboxane (TX) levels and plasma LDL cholesterol or beta thromboglobulin (beta-TG) levels (r = 0.48, P < 0.05: r = 0.47, P < 0.05, respectively), whereas, at peak exercise, TX and beta-TG levels increased, but no correlation was seen. At peak exercise, platelets showed hyperaggregability in terms of maximal amplitude and reaction slope (P < 0.001 and P < 0.01, respectively). In contrast to the increase in plasma lipid peroxide levels seen during peak exercise (P < 0.05), LDL lipid peroxides decreased during exercise, this decrease reaching a statistical significance at 10 min postexercise (P < 0.05). In addition, the ex vivo addition of mildly oxidized LDL (10 mg protein x L(-1)) to peak exercise blood resulted in a significant attenuation of platelet aggregation and a decrease in TX release. At 10 min postexercise, a correlation was seen between LDL lipid peroxides and TX levels (r = 0.78, P < 0.001) or beta TG levels (r = 0.68, P < 0.005). CONCLUSION: These results suggest that LDL lipid peroxides play a role in modulating and attenuating platelet aggregation during strenuous exercise. PMID- 10795784 TI - Carbohydrate availability affects ammonemia during exercise after beta 2 adrenergic blockade. AB - PURPOSE: Beta-adrenergic blockade increases blood ammonia concentration during exercise. The purpose of this study was to assess the role of decreased carbohydrate availability in this process. METHODS: Wistar rats (N = 47) were injected intravenously with a selective beta 2-adrenoceptor blocker (ICI 118,551), placebo, or beta 2-blocker + glucose 1 h before a treadmill exercise test. Blood samples were taken to measure the concentration of ammonia, glucose, lactic acid, free fatty acids (FFA), glycerol, branched-chain amino acids (BCAA), and muscle samples for determination of glycogen content. RESULTS: Beta 2 adrenergic blockade shortened running time to exhaustion (23 +/- 4.3 min compared to 44 +/- 5.2 min with placebo), increased blood ammonia levels (146.7 +/- 16.21 micromol x L(-1) compared to 47.5 +/- 0.92 micromol x L(-1) with placebo) and prevented exercise-induced glycogen breakdown in soleus and gastrocnemius muscles. Pre-exercise supplementation of glucose during beta 2-blockade restored exercise-induced glycogen breakdown and reduced blood ammonia concentration during exercise (66.5 +/- 5.65 mmol x L(-1)) but did not improve exercise capacity (26 +/- 3.2 min) when compared with beta2-blockade alone. CONCLUSION: The results suggest that the enhanced rise in blood ammonia concentration during exercise after beta-blockade is caused by impaired carbohydrate availability. PMID- 10795785 TI - Testosterone-propionate impairs the response of the cardiac capillary bed to exercise. AB - OBJECTIVE: Experimental application of anabolic-androgenic steroids and exercise training induce cardiac hypertrophy. This study quantifies for the first time, on microscopical level, the adaptation of the cardiac capillaries and myocytes to the concomitant application of testosterone-propionate and exercise training. METHODS: Female SPF-NMRI mice were studied over 3 and 6 wk. Experimental groups: (i) sedentary control (C); (ii) exercise (treadmill running, E); (iii) testosterone-propionate (TP); and (iv) testosterone-propionate+exercise (TPE). Morphometric parameters: 1) papillary muscles: capillary density, intercapillary distance, number of capillaries around a myocyte, and minimal myocyte diameter; and 2) left ventricular wall: capillary density and intercapillary distance. RESULTS: Papillary muscle: A striking suppression of the exercise-induced improvement in capillary supply occurs in the testosterone-propionate+exercise groups over 3 and 6 wk. Exercise without drugs increases significantly (P < 0.05) the capillary density, shortens significantly (P < 0.05) the intercapillary distance, whereas it increases the number of capillaries around a myocyte. These alterations are not observed in the testosterone-propionate treated sedentary animals; e.g., capillary density after 6 wk (mean values +/- standard deviation, capillaries x mm(-2)): C: 4272 +/- 287, E: 5411 +/- 758, TP: 4221 +/- 364, and TPE: 3997 +/- 397. Moreover, only in the testosterone-propionate+exercise groups occurs a mild myocyte hypertrophy after both time periods: there is a trend toward hypertrophy (P < 0.1) in comparison with the C groups and a significant hypertrophy (P < 0.05) in comparison with the E groups. CONCLUSIONS: Testosterone propionate profoundly inhibits the exercise-induced augmented capillarization, whereas (under training conditions) it leads to a mild myocyte hypertrophy. The microvascular impairment could trigger an imbalance between the myocardial oxygen supply and demand, especially during physical exercise. PMID- 10795786 TI - Influence of acute and chronic treadmill exercise on rat brain POMC gene expression. AB - PURPOSE AND METHODS: In situ hybridization using biotin-labeled POMC cRNA probes and integrated image pattern analysis were used to observe the effect of chronic (total 7 wk) and acute treadmill exercise (an initial speed of 15 m x min(-1) gradually increased to 35 m x min(-1) with 0 degrees, 20-25 min x d(-1) duration) on the changes of POMC mRNA expression in different areas of the rat brain. RESULTS: Acute exercise performed after 7 wk of training resulted in a decrease in the expression of POMC mRNA in both the frontal cortex and hippocampal CA1 of the experimental rats. Three hours postexercise, the POMC mRNA indicator had not yet returned to normal expression levels in the frontal cortex, but hippocampal CA1 levels showed increases above normal levels 30 min postexercise. Expression levels in the hypothalamus were shown to be increased immediately after acute exercise and remained high even during remeasurement 3 h postexercise. CONCLUSION: Changes in POMC gene expression in response to chronic and high intensity acute exercise vary depending on the brain region examined. PMID- 10795787 TI - Effect of acute plasma volume expansion on thermoregulation and exercise performance in the heat. AB - PURPOSE: We investigated the effects of acute plasma volume expansion on exercise performance in the heat. METHODS: Six moderately trained men cycled for 40 min at 64 +/- 2% peak pulmonary oxygen uptake (VO2peak) followed by an individual performance time trial, where subjects completed a set amount of work (267 +/- 15 kJ) in as little time as possible. Exercise trials were performed at 35 degrees C with a relative humidity of 40%. Subjects performed two exercise trials: one after 13.1 +/- 1% acute plasma volume expansion (PVE), which was achieved by the intravenous infusion of 8 mL x kg(-1) body weight of Hemaccel (35 g x L(-1) polygeline, 145 mmol x L(-1) Na+, and 145 mmol x L(-1) Cl-) and the other without prior treatment (CON). RESULTS: Core temperature, skin blood flow, and heart rate progressively increased (P < 0.05) during exercise, but no differences were observed between trials. Plasma glucose and lactate were similar at rest and during exercise, as was VO2 during exercise. Exercise performance was not influenced by plasma volume expansion (CON 17.5 +/- 0.4 min and PVE 17.1 +/- 0.2 min). CONCLUSION: These data suggest that, in moderately trained men, plasma volume expansion alone does not enhance thermoregulatory function and exercise performance during moderate intensity exercise in the heat. PMID- 10795788 TI - A review of correlates of physical activity of children and adolescents. AB - BACKGROUND: Understanding the factors that influence physical activity can aid the design of more effective interventions. Previous reviews of correlates of youth physical activity have produced conflicting results. METHODS: A comprehensive review of correlates of physical activity was conducted, and semiquantitative results were summarized separately for children (ages 3-12) and adolescents (ages 13-18). The 108 studies evaluated 40 variables for children and 48 variables for adolescents. RESULTS: About 60% of all reported associations with physical activity were statistically significant. Variables that were consistently associated with children's physical activity were sex (male), parental overweight status, physical activity preferences, intention to be active, perceived barriers (inverse), previous physical activity, healthy diet, program/facility access, and time spent outdoors. Variables that were consistently associated with adolescents' physical activity were sex (male), ethnicity (white), age (inverse), perceived activity competence, intentions, depression (inverse), previous physical activity, community sports, sensation seeking, sedentary after school and on weekends (inverse), parent support, support from others, sibling physical activity, direct help from parents, and opportunities to exercise. CONCLUSION: These consistently related variables should be confirmed in prospective studies, and interventions to improve the modifiable variables should be developed and evaluated. PMID- 10795789 TI - Comparing physical activity assessment methods in the Seasonal Variation of Blood Cholesterol Study. AB - PURPOSE: This paper evaluated three measures of physical activity employed in the Seasonal Variation of Blood Cholesterol Study (Seasons), and it had two objectives: 1) To examine the laboratory validity of the Actillume activity monitor, and 2) To examine the relative validity of three 24-h physical activity recalls (24HR) in quantifying short-term physical activity behaviors. METHODS: Nineteen healthy middle-age adults completed seven activity trials (reading, typing, box moving, stepping, and walking (3.5, 4.25, 5.0 km x h(-1))) while oxygen consumption and Actillume measures were obtained. ANOVA, linear regression, and a scatter plot were employed to examine the validity of the Actillume. In relative validity analyses of the 24HR in the Seasons study, participants (N = 481) completed two or three 24HR (MET-h x d(-1)) and a modified Baecke Questionnaire. A subset of the cohort (N = 41) wore the Actillume for 3-8 d (counts x min(-1) x d(-1)). The relative validity of the 24HR method was examined by comparison to these criterion measures. RESULTS: In laboratory validation analyses, the monitor was found to discriminate between sedentary and moderate intensity activities, changes in walking speed, and to account for 79% of the variance in oxygen consumption across sedentary and walking trials. In relative validity analyses, correlations between the 24HR and the modified Baecke ranged from 0.29 to 0.52 (P < 0.01) across total, household, occupational, and leisure-time activities. CONCLUSIONS: In laboratory testing, the Actillume monitor discriminated between sedentary and moderate intensity activities and was highly correlated with oxygen consumption. Three 24HR of physical activity were observed to have a relative validity that was comparable to published data from other short-term activity assessments that also employed the Baecke Questionnaire and activity monitors as criterion measures. PMID- 10795790 TI - Knee strength variability between individuals across ranges of motion and hip angles. AB - PURPOSE: Isokinetic strength is normally measured for a single range of motion and body position. This study quantified the variability, between individuals, in the relationships between a single peak knee extension moment and the isokinetic extension moments measured for different hip angles and ranges of knee motion. Effects of hip angle, and of the starting knee angle of the range of motion, on isokinetic knee extension strength were also determined. METHODS: The isokinetic knee extension strength of 10 subjects was measured at 30 degrees x s(-1) to a knee flexion angle of 10 degrees from starting knee angles of 90, 75, 60, 45, and 30 degrees, in both the seated and supine positions. Moments were normalized to the peak moment from a reference contraction. RESULTS: Peak moments and moments at larger knee flexion angles were greater in the seated than in the supine position. The starting knee angle affected the peak moment, the angle of peak moment, and the moments over the initial and final portions of the range of motion. Peak moments were highly correlated between all hip angle-starting knee angle combinations. However, the normalized peak moments, the angles of peak moment, and the normalized angle-specific moments all varied considerably between subjects. The pooled standard deviation and average coefficient of variation of the normalized angle-specific moments between subjects were 10.5% of the normalizing moment and 15.7%, respectively. Excluding the reference contraction, between-subject variability was unaffected by hip angle or starting knee angle. CONCLUSIONS: Influences of hip angle, starting knee angle, and individual differences on isokinetic knee extension strength must be considered to ensure that the moments obtained from isokinetic testing adequately reflect the general strength capabilities of an individual. PMID- 10795791 TI - Effect of oral creatine supplementation on isokinetic torque production. AB - PURPOSE: This study was conducted to examine the effect of oral creatine supplementation on the decline in peak isokinetic torque of the quadriceps muscle group during an endurance test. METHODS: Twenty-three active, but untrained, male subjects performed isokinetic strength tests on a Cybex II dynamometer at 180 degrees x s(-1). The protocol consisted of pre- and post-tests with five sets of 30 maximum volitional contractions with a 1-min rest period between sets. Subjects returned to perform the posttest after 5 d of placebo (4 x 6 g glucose x d(-1), N = 12) or creatine (4 x 5 g creatine + 1 g glucose x d(-1), N = 11) supplementation. Supplements and testing were administered in a double blind fashion. Peak torque was measured during each contraction and the 30 contractions were averaged for each set. RESULTS: A three-way mixed ANOVA with one between factor (placebo vs creatine) and two within factors (pre/post supplementation and sets 1-5) revealed no significant interactions, P > 0.05. The placebo vs creatine main effect was also nonsignificant, whereas the pre/post and set effects were significant (P < 0.05). Peak torque increased (approximately 3%) from pre- to post-testing, (P = 0.04), but the absolute magnitude of the differences is unlikely to be of any practical significance. Peak torque decreased from sets 1 to 4, whereas sets 4 and 5 were not different. A priori contrasts comparing the creatine group's performance pre vs post test for the fourth and fifth sets were nonsignificant (P > 0.05). CONCLUSIONS: Based on within and between group comparisons, we were unable to detect an ergogenic effect of oral creatine supplementation on the decline in peak torque during isokinetic exercise at 180 degrees x s(-1). PMID- 10795792 TI - Consistency of swimming performance within and between competitions. AB - PURPOSE: The consistency of performance between events impacts how athletes should specialize in events, how competitions should be structured, and how changes in performance affect an athlete's placing in an event. We have therefore determined the consistency of swimming performance in events within and between two national-level competitions. METHODS: We used mixed linear modeling to analyze official performance times of 149 male and 162 female swimmers at a junior national championship, and of 117 male and 104 female swimmers at an open national championship 20 d later. The events differed in stroke (backstroke, breaststroke, butterfly, freestyle, and individual medley) or distance (50-1500 m). RESULTS: Swimmers were most consistent in their performance for the same event between the two competitions (typical variation between competitions, 1.4%; 95% likely range of true value, 1.3-1.5%). They were less consistent between distances of a given stroke within each competition (1.7%; 1.5-1.9%) and least consistent between strokes for a given distance (2.7%; 2.3-3.1%). Variation in performance between the longest continuous freestyle distances (400, 800, and 1500 m) in the open competition was half that between widely spaced freestyle distances (50, 200, and 800 m). Faster swimmers were more consistent (1.1%; 0.9 1.4%) for the same event between competitions than slower swimmers (1.5%; 1.3 1.9%). CONCLUSIONS: (a) Swimmers are stroke specialists rather than distance specialists; with the present set of events in competitions, they should concentrate training and competing on a particular stroke rather than a particular distance. (b) More swimmers would have a chance of winning a medal if events of a given stroke differed more widely in distance. (c) Factors that affect performance time by as little as 0.5% will affect the placing of a top junior swimmer. PMID- 10795793 TI - Intensity of exercise during road race pro-cycling competition. AB - PURPOSE: The aim of this study was to quantify the intensity of competition during two professional bicycle stage races: the Tour de France (Tour) and Vuelta a Espana (Vuelta). METHODS: The HR responses of 18 world class cyclists were recorded during the races and compared with HR ranges that corresponded to four intensities of exercise that were measured in the laboratory with an incremental test to exhaustion 2 wk before each race. The four intensities were: Anaerobic (AN) over the individual anaerobic threshold, which was over 90% of VO2max; intense aerobic (IA), which was between 70 and 90% of VO2max; moderate aerobic (MA), which was between 50 and 70% of VO2max; and recovery (RE), which was < 50% of VO2max. The stages were divided in individual time trial (ITT), flat, or mountain. RESULTS: The mean HR of the Vuelta and Tour were, respectively, 133.8 +/- 17.9 and 134 +/- 18.6 beats x min(-1). The mean total time of each stage was 269.6 +/- 122 and 259.4 +/- 119.9 min. The mean stage time over IAT was 17.5 +/- 15.7 and 24.7 +/- 26 min; the IA time was 75.2 +/- 47.6 and 79.6 +/- 48.3 min; the MA was 97.2 +/- 57.4 and 89.5 +/- 54.9 min. Finally the RE time was 79.6 +/- 60.5 and 65.4 +/- 69.7 min. The percentage of participation related to total time of the race was, respectively, in the Vuelta and the Tour, 12.99 and 16.8% in AN exercise intensity, 29.5 and 29.2% in IA, 32.4 and 31.9% in MA, and 25.1 and 25.2% in RE. There are no differences in AN time among flat, mountain, and ITT stages in each race, except for the mountain stages in the Tour. CONCLUSION: Cycling is a high intensity sport because approximately 93 min in flat and 123 min in mountain stages were above 70% of VO2max. In addition, the time spent at IAT was roughly 20 min regardless of stage type, suggesting that the anaerobic capacity limits performance. PMID- 10795794 TI - Validity and factorial invariance of the Social Physique Anxiety Scale. AB - PURPOSE: The present study 1) tested whether the two-factor model to the 12-item Social Physique Anxiety Scale (SPAS) was substantively meaningful or a methodological artifact representing positively and negatively worded items, 2) assessed the factorial validity of the nine-item unidimensional model to the SPAS, 3) examined whether modifying the number of SPAS items would improve the factorial validity. 4) evaluated the factorial invariance of the SPAS across gender, and 5) explored the construct validity of SPAS scores. METHODS: Female (N = 146) and male (N = 166) college students (22.2 +/- 4.0 yr) in lecture (N = 103) and physical activity (N = 209) courses completed the SPAS, Physical Self Efficacy Scale (PSES), Surveillance subscale of the Objectified Body Consciousness Scale (S-OBCS), and short form of the Marlowe-Crowne Social Desirability Scale (SDS-C). RESULTS: Confirmatory factor analyses (CFA) revealed that the two-factor model to the 12-item SPAS was a methodological artifact representing positively and negatively worded items. CFA indicated that the nine item unidimensional model represented an acceptable fit to the SPAS, but it also could be improved. Modifications based on standardized residuals and item content led to the removal of two items and a seven-item unidimensional solution to the SPAS. The nine- and seven-item models demonstrated factorial invariance across gender. Correlation analyses between nine- and seven-item SPAS scores to PSES, S OBCS, and SDS-C provided support for the construct validity. CONCLUSIONS: The nine- and seven-item unidimensional models to the SPAS demonstrated evidence of factorial validity, factorial invariance, and construct validity; the two-factor model to the SPAS represented a methodological artifact. PMID- 10795795 TI - Measurement of daily walking distance-questionnaire versus pedometer. AB - PURPOSE: This study compared measurements of daily walking distance on the College Alumnus questionnaire (CAQ) and a pedometer. METHODS: A total of 96 men and women (25-70 yr of age) with a wide range of physical activity habits were studied. Physical activity index was computed from the College Alumnus questionnaire (PAI-CAQ) as the sum of the energy expended in stair climbing, walking, and sports and recreational physical activity. Data on walking distance were compared with values obtained from the Yamax electronic pedometer (DW-500B). Participants wore the pedometer for 7 consecutive days, except when sleeping, showering, or performing sports and recreational activities. RESULTS: Subjects underestimated their daily walking distance on the CAQ, compared with the pedometer (1.43 +/- 1.01 vs 4.17 +/- 1.61 km x d(-1)). The energy expended in walking was correspondingly lower on the CAQ, compared with the pedometer-derived values (555 +/- 405 versus 1608 +/- 640 kcal x wk(-1)). CONCLUSIONS: These findings suggest that electronic pedometers are useful for examining questions about walking distance on physical activity questionnaires. PMID- 10795796 TI - Determination of circulating hemoglobin mass and related quantities by using capillary blood. AB - PURPOSE: A standardized carbon monoxide (CO) rebreathing procedure with measurements of CO-hemoglobin, hemoglobin concentration ([Hb]), and hematocrit (Hct) enables to determine total Hb mass (Hb(tot)), blood, erythrocyte, and plasma volume (BV, EV, and PV). These calculations are normally based on venous blood samples. However, micromethods also allow determinations from capillary blood. METHODS: The accuracy of using capillary blood for Hb(tot), BV, EV, and PV determination was evaluated in 42 men (age: 25.1 +/- 4.0 yr, body mass: 80.3 +/- 9.6 kg) by comparison of capillary and venous data. RESULTS: Capillary Hb(tot) (962 +/- 110 g) did not differ from venous values (959 +/- 106 g). Hb(tot) values were highly correlated (r = 0.987, P < 0.001, SEE 18 g). Also, capillary and venous BV, PV, and EV were highly correlated (0.94 < r < 0.98), but slightly different (-2.7 to 0.9%) because of higher capillary than venous [Hb] and Hct. Coefficients of variation of repeated Hb(tot), EV, PV, and BV measurements (3.0 5.2%) were similar in capillary and venous blood. CONCLUSION: Calculation of Hb(tot) using capillary blood is as accurate and reliable as using venous blood. PMID- 10795797 TI - Comparison of cardiac output measured by two automated methods of CO2 rebreathing. AB - PURPOSE: The aim of the present study was to investigate the reproducibility of the exponential method of CO2 rebreathing with the use of automated curve fitting and to determine whether this method is superior to the equilibrium method in terms of reproducibility and clinical practicability. METHODS: Repeated measurements of cardiac output were performed using the automated equilibrium and exponential methods. These measurements were compared in 12 healthy male subjects at rest and during incremental exercise tests. RESULTS: Estimated cardiac output was not significantly different between duplicate measurements at rest nor at any level of exercise with either method. At rest the exponential method showed a tendency toward larger variability than the equilibrium method. The exponential method produced significantly higher (P < or = 0.001) estimates at rest (averaging up to 9.8 L x min(-1)) compared with the equilibrium method (averaging up to 6.5 L x min(-1)). Reproducibility improved for both methods with increasing workloads, and a second measurement at rest also seemed more reproducible and valid than the first. During exercise, both methods produced comparable values for cardiac output, and highly significant relations between cardiac output and oxygen uptake were observed for both methods (ranging from r2 = 0.79 to r2 = 0.88, P < or = 0.001). The equilibrium method produced unpleasant side effects more frequently (75% vs 21%, P < or = 0.001) compared with the exponential method and lead more subjects to premature interruption of the exercise test because of the rebreathing maneuver (42% vs 17%, P = 0.058). CONCLUSIONS: Automated curve fitting for the exponential method gave reproducible and valid results during submaximal and maximal exercise but not at rest. The equilibrium method on the other hand interfered with exercise. Therefore, the equilibrium method is recommended at rest and at lower levels of exercise and the exponential method at higher intensities. PMID- 10795798 TI - What has changed? PMID- 10795799 TI - Rigid contact lens fitting relationships in keratoconus. PMID- 10795800 TI - Acute, idiopathic blind spot enlargement syndrome. AB - An otherwise healthy 12-year-old boy was evaluated for an enlarged blind spot in his left eye. Neither optic nerve edema/neuritis nor a retrobulbar mass explained this finding. Consultation with a neuro-ophthalmologist over a period of 14 months resulted in a diagnosis of acute, idiopathic blind spot enlargement syndrome, a rare and poorly understood ocular condition. To the author's knowledge, this is the youngest case ever reported. PMID- 10795801 TI - Preoptometry and optometry school grade point average and optometry admissions test scores as predictors of performance on the national board of examiners in optometry part I (basic science) examination. AB - PURPOSE: To evaluate preoptometry and optometry school grade point averages and Optometry Admission Test (OAT) scores as predictors of performance on the National Board of Examiners in Optometry NBEO Part I (Basic Science) (NBEOPI) examination. METHODS: Simple and multiple correlation coefficients were computed from data obtained from a sample of three consecutive classes of optometry students (1995-1997; n = 278) at Southern California College of Optometry. RESULTS: The GPA after year two of optometry school was the highest correlation (r = 0.75) among all predictor variables; the average of all scores on the OAT was the highest correlation among preoptometry predictor variables (r = 0.46). Stepwise regression analysis indicated a combination of the optometry GPA, the OAT Academic Average, and the GPA in certain optometry curricular tracks resulted in an improved correlation (multiple r = 0.81). Predicted NBEOPI scores were computed from the regression equation and then analyzed by receiver operating characteristic (roc) and statistic of agreement (kappa) methods. From this analysis, we identified the predicted score that maximized identification of true and false NBEOPI failures (71% and 10%, respectively). Cross validation of this result on a separate class of optometry students resulted in a slightly lower correlation between actual and predicted NBEOPI scores (r = 0.77) but showed the criterion-predicted score to be somewhat lax. CONCLUSIONS: The optometry school GPA after 2 years is a reasonably good predictor of performance on the full NBEOPI examination, but the prediction is enhanced by adding the Academic Average OAT score. However, predicting performance in certain subject areas of the NBEOPI examination, for example Psychology and Ocular/Visual Biology, was rather insubstantial. Nevertheless, predicting NBEOPI performance from the best combination of year two optometry GPAs and preoptometry variables is better than has been shown in previous studies predicting optometry GPA from the best combination of preoptometry variables. PMID- 10795802 TI - Repeatability of the Visual Manipulation Test. AB - BACKGROUND: The Getman-Henderson-Marcus Visual Manipulation Test (VMT) is a commonly used test of visualization with an objective scoring procedure. However, its repeatability has not been studied. PURPOSE: The purpose of this investigation was to assess the repeatability of the VMT in children in the second through fourth grades. METHOD: The VMT was administered to the children (n = 193; mean age, 8.72 years) from a middle class elementary school near Columbus, Ohio. One class from each grade (n = 56) was retested by the same optometrist within 1 month. Repeatability analysis included plotting the difference vs. the mean of the scores obtained at the initial test and retest. RESULTS: All the children were able to complete the test. Means and SD's were found to be 5.23+/ 1.98 for second graders, 5.76+/-2.10 for third graders, and 6.95+/-2.30 for fourth graders. The mean difference between test and retest scores was close to zero. The 95% limits of agreement were found to be -4.5 to 5.2. CONCLUSION: No consistent learning effect seemed to be present upon retest. However, analysis revealed poor repeatability for the VMT. Therefore, the VMT should not be used to monitor progress with therapy. PMID- 10795803 TI - Longitudinal changes in corneal asphericity in myopia. AB - PURPOSE: To report the change in shape of the peripheral cornea (asphericity, Q) as it relates to myopia progression in adolescence. METHODS: Forty-eight subjects with initial ages between 11 and 13 years of age were observed for 5 years. Each subject had participated in a variety of soft contact lens studies and all had worn daily wear soft contact lenses successfully for the 2 years before collection of the second data set. RESULTS: Myopia increased by an average of 1.46 D and was strongly correlated with a 0.56-mm increase in axial length. There was no contribution to the change in myopia from the change in central radius of the cornea. A significant (p < 0.01, one tail) correlation was found between Q and the progression of myopia. Principal axis analysis yielded a slope of -0.04 in Q per diopter of increase in myopia. CONCLUSION: These data show a shift to a more positive Q (in the oblate direction) with increased myopia. PMID- 10795804 TI - Age-related changes in human ciliary muscle. AB - PURPOSE: To examine the role of the ciliary muscle in accommodation and presbyopia. METHODS: Sixteen pairs of eyes from donors aged 1 to 107 years were treated with atropine or pilocarpine and then processed for light microscopy. Seven pairs were analyzed for treatment effects, including morphometric measurements of muscle dimensions, muscle fiber group area, and the percentage of connective tissue. RESULTS: The ciliary muscle shortened in length in response to drug treatment at all ages. The ciliary muscle of older subjects contained greater amounts of connective tissue and was shorter, wider and the internal apical edge moved forward. CONCLUSIONS: This study confirms previous reports that the human ciliary muscle retains its ability to contract throughout the lifespan. However, the ciliary muscle also displayed age-related changes, which may be partly accounted for by the forces exerted from the aging lens and zonules. PMID- 10795805 TI - A new method for determining prismatic effects in cylindrical spectacle corrections. AB - This study presents a new method of finding differential prismatic effects in an anisometropic spectacle correction containing cylindrical lenses. The calculations are based on dynamic spectacle magnifications rather than Prentice's rule. If an imaginary circular object is considered, cylindrical lenses will produce elliptical far point images, which can be superimposed on the spectacle plane for comparison. The difference between left and right ellipses, in any meridian, then represents the distance the eyes have to diverge in order to fuse the object of regard. This distance can be translated into prism diopters of differential prismatic effect. Although the conventional methods for finding this effect often result in very large errors, the new method can be performed with great accuracy. In part, this is because it uses the actual eccentricities of the two eyes rather than an assumed average eccentricity. Moreover, the method includes considerations of base curves and center thickness. Contrary to the classical methods, it can thus be applied to clinically realistic lenses. PMID- 10795806 TI - Independent synthesis of aminophospholipid-linked maillard products. AB - Phospholipid-linked glycation products are supposed to play an important role in lipid oxidation in vivo. Independent syntheses and unequivocal structural characterization are reported for the phosphatidyl ethanolamine (PE)-derived Amadori compound 4-hydroxy-4-oxo-1-[(palmitoyloxy)methyl]-9-(2,3,4,5-tetrahydrox ytetrahydro-2H-pyran-2-yl)-3,5-dioxa-8-aza-4lambda5-ph osphanon-1-yl palmitate, pyrrolecarbaldehyde 2-[[[2-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1 yl]ethoxy](hydroxy)phosph oryl]oxy]-1-[(palmitoyloxy)methyl]ethyl palmitate, the carboxymethyl (CM) derivative 7-hydroxy-7,13-dioxo-10-(palmitoyloxy)-6,8,12 trioxa-3-aza-+ ++7lambda5-phosphaoctacosan-1-oic acid, and the carboxyethyl (CE) derivative 7-hydroxy-2-methyl-7,13-dioxo-10-(palmitoyloxy)-6,8,12-trioxa++ +-3 aza-7lambda5-phosphaoctacosan-l-oic acid. With these reference compounds, a liquid chromatography-mass spectrometry (LCMS) method for the determination of such PE-linked Maillard products has been developed. PMID- 10795807 TI - Highly efficient synthesis of 1-thioglycosides in solution and solid phase using iminophosphorane bases. AB - Disaccharides of 1-thioglycosides, an important class of glycomimics, can be synthesized by direct S-alkylation in exceptionally high yields when iminophosphorane bases are employed. The reaction conditions employed appear to be general and stereospecific. Axial and equatorial 4-triflates and primary tosylates of alkyl pyranosides provided excellent yields of thio-disaccharides without substantial elimination products. The iminophosphorane bases also proved to be useful in solid support-bound couplings of thioglycosides though with lower efficiency. PMID- 10795808 TI - The conformation of alpha-(1-->4)-linked glucose oligomers from maltose to maltoheptaose and short-chain amylose in solution. AB - The conformation of maltose-type oligomers in water and in dimethylsulfoxide (Me2SO) was studied using two-dimensional NMR spectra. In Me2SO all of the oligomers have a 1a-type conformation. In water, they tend to adopt the same conformation, but the oligomers are looser and more flexible than in Me2SO. PMID- 10795809 TI - Distribution of methyl substituents in amylose and amylopectin from methylated potato starches. AB - Granular potato starches were methylated in aqueous suspension with dimethyl sulfate to molar substitution (MS) values up to 0.29. Fractions containing mainly amylose or amylopectin were obtained after aqueous leaching of the derivatised starch granules. Amylopectin in these fractions was precipitated with Concanavalin A to separate it from amylose. Amylose remained in solution and was enzymatically converted into D-glucose for quantification, thereby taking into account the decreased digestibility due to the presence of methyl substituents. It was found that the MS of amylose was 1.6-1.9 times higher than that of amylopectin in methylated starch granules. The distributions of methyl substituents in trimers and tetramers, prepared from amylose- or amylopectin enriched fractions, were determined by FAB mass spectrometry and compared with the outcome of a statistically random distribution. It turned out that substituents in amylopectin were distributed heterogeneously, whereas substitution of amylose was almost random. The results are rationalised on the basis of an organised framework that is built up from amylopectin side chains. The crystalline lamellae are less accessible for substitution than amorphous branching points and amylose. PMID- 10795810 TI - Full assignment of the proton and carbon NMR spectra and revised structure for the capsular polysaccharide from Streptococcus pneumoniae type 17F. AB - Full proton, 13C and 31P NMR assignments for the capsular polysaccharide from Streptococcus pneumoniae Type 17F are reported, and a revised structure differing in the anomeric configuration of the sidechain beta-Galp residue proposed. This polysaccharide is a component of the current 23-valent polysaccharide vaccine. The implications of this revised structure for published work are discussed. PMID- 10795811 TI - Identification and synthesis of a trisaccharide produced from lactose by transgalactosylation. AB - Enzymatic transgalactosylation of lactose by means of Streptococcus thermophilus, subspecies DN-001065, led to a mixture of D-galactose (approximately 4%), D glucose (approximately 15%), lactose (approximately 51%), minor disaccharides (6%), trisaccharides (approximately 20%) and tetrasaccharides (3%). The major trisaccharide (approximately 16%) was identified by NMR spectroscopy and chemical synthesis as being the known beta-D-galactopyranosyl-(1-->3)-beta-D galactopyranosyl-(1-->4)-D-glucos e (3'-beta-D-galactopyranosyl-lactose). It was purified from a mixture of peracetylated oligosaccharides by column chromatography followed by deacetylation. For the first time, 3'-beta-D galactopyranosyl-lactose has been obtained on the 1 g scale, by resorting to simple techniques and equipment. NMR spectra have been unambiguously assigned. PMID- 10795812 TI - Synthesis of a chitosan tetramer derivative, beta-D-GlcNAc-(1-->4)-beta-D-GlcNAc (1-->4)-beta-D-GlcNAc-(1-->4)-D-Glc N through a partial N-acetylation reaction by chitin deacetylase. AB - We have synthesized beta-D-GlcNAc-(1-->4)-beta-D-GlcNAc-(1-->4)-beta-D-GlcNAc-(1- >4)-D-GlcN (2) through a partial N-acetylation reaction of chitosan tetramer 1 by a chitin deacetylase from Colletotrichum lindemuthianum ATCC 56676. The compound was purified from the mixture of acetylation products of 1 using cation-exchange column chromatography and amine-adsorption column chromatography, and its structure was estimated by 1H NMR and FABMS analyses. The enzymatic reaction allows a regioselectivity that is hard to achieve by chemical N-acetylation. PMID- 10795814 TI - Structure elucidation of the O-chain from the major lipopolysaccharide of the Xanthomonas campestris strain 642. AB - A novel O-polysaccharide consisting of D-Xylp and L-Rhap in the molar ratio of 1:2.5 was identified as the major component in the lipopolysaccharide fraction of Xanthomonas campestris strain 642, which is responsible for a new bacterial disease of the strawberry plant. Its structure was mainly determined using chemical analysis, Smith degradation and 1D and 2D NMR spectroscopy experiments as: carbohydrate sequence [see text]. PMID- 10795813 TI - Isolation and characterization of non-labeled and 13C-labeled mannans from Pichia pastoris yeast. AB - Mannans from genetically modified Pichia pastoris yeast, used for overproduction of neural cell adhesion molecule protein, grown on normal media or on uniformly 13C-labeled glucose and methanol, were isolated and characterized by high-field (750 MHz) NMR spectroscopy. Fully 13C-labeled oligosaccharide fragments were prepared from mannans by acetolysis. According to the data obtained, the mannan is made up of a main chain of alpha-(1-->6)-linked mannopyranosyl residues, substituted at 0-2 with alpha-mannopyranosyl or a alpha-D-Manp-(1-->2)-beta-D Manp-(1-->2)-beta-D-Manp-( 1-->2)-alpha-D-Manp- group, and with much lower content of substitution with beta-D-Manp-(1-->2)-alpha-D-Manp-. A fraction of these oligosaccharide side chains is again substituted with alpha-D-Glcp or alpha D-GlcpNAc through a phosphodiester linkage to the 6 position of the first mannopyranosyl residue. Improved conditions of acetolysis, cleaving all alpha-(1- >6) linkages, but not beta-mannoside linkages, are proposed. PMID- 10795815 TI - Isolation of hemicellulose from a sorghum, Andropogon sorghum Brot, Kumadake no. 263, and determination of its constituent sugars. AB - Hemicellulose was isolated from the stem of sorghum, Andropogon sorghum Brot, Kumadake no. 263, and the constituent sugars of this isolate were determined. The xylose concentration in the hemicellulose of SV263 was extremely high. PMID- 10795816 TI - Synthesis of a model compound related to an anti-ulcer pectic polysaccharide. AB - A stereocontrolled synthesis of the model compound for an anti-ulcer active polysaccharide (Bupleuran 2IIc) is described. Glycosidation of the disaccharide acceptor, 2-O-acetyl-3-O-benzyl-4-O-(p-methoxybenzyl)-alpha-L-rhamnopyranosyl-(1- >4)-2,3,6-tri-O-benzyl-alpha-D-galactopyranosyl trichloroacetimidate, with the disaccharide receptor, allyl 3,4-di-O-benzyl-alpha-L-rhamnopyranosyl-(1-->4) 2,3,6-tri-O-benzyl-beta- D-galactopyranoside, using silver triflate (AgOTf) as a promoter gave the desired tetrasaccharide derivative, which was transformed into the acidic tetrasaccharide, corresponding to a segment of the rhamnogalacturonan (Bupleuran 2IIc) polysaccharide, propyl alpha-L-Rha-(1-->4)-alpha-D-GalA-(1-->2) alpha-L-Rha-(1-->4)-beta-D-GalA , via removal of the corresponding ether and ester protecting groups, followed by oxidation. PMID- 10795817 TI - Unexpected stereochemical outcome of activated 4,6-O-benzylidene derivatives of the 2-deoxy-2-trichloroacetamido-D-galacto series in glycosylation reactions during the synthesis of a chondroitin 6-sulfate trisaccharide methyl glycoside. AB - The synthesis of methyl (beta-D-glucopyranosyluronic acid)-(1-->3)-(2-acetamido-2 deoxy-6-O-sulfonato-beta-D-galactopyr anosyl)-(1-->4)-(beta-D glucopyranosid)uronate trisodium salt, a chondroitin 6-sulfate trisaccharide derivative, is described. Loss of stereocontrol in glycosylation reactions involving activated 4,6-O-benzylidene derivatives of the 2-deoxy-2 trichloroacetamido-D-galacto series and D-glucuronic acid-derived acceptors was highlighted. This draw-back was overcome through the use of phenyl 3,4,6-tri-O acetyl-2-deoxy-1-thio-2-trichloroacetamido-beta-D-gala ctopyranoside, which afforded the desired beta-linked disaccharide derivative in high yield with an excellent stereoselectivity. This later was submitted to acid-catalyzed methanolysis, followed by benzylidenation, and condensed with methyl 2,3,4-tri-O benzoyl-1-O-trichloroacetimidoyl-alpha-D-glucopyran uronate to afford the expected trisaccharide derivative. Subsequent transformation of the N trichloroacetyl group into N-acetyl, mild acid hydrolysis, selective O sulfonation at C-6 of the amino sugar moiety, and saponification afforded the target molecule as its sodium salt in high yield. PMID- 10795818 TI - Preparation of selenoanhydro- and telluroanhydroglycofuranosides and some corresponding nucleosides. AB - Methyl 2,3-anhydro-alpha-D-ribofuranoside (3a) was transformed into methyl 2 seleno-2,5-anhydro-alpha-D-arabinofuranoside (5a) and methyl 3-seleno-3,5-anhydro alpha-D-xylofuranoside (6a) in two steps via the reaction of the C-5 mesylate of 3a, methyl 2,3-anhydro-5-O-mesyl-alpha-D-ribofuranoside (4a), with sodium hydrogen selenide. The corresponding beta anomer of 3a yielded methyl 3-seleno 3,5-anhydro-beta-D-xylofuranoside as the main product and only traces of methyl 2 seleno-2,5-anhydro-beta-D-arabinofuranoside. Sodium hydrogen telluride transformed 4a into methyl 2-telluro-2,5-anhydro-alpha-D-arabinofuranoside. Starting from 5a we prepared 1-(2-seleno-2,5-anhydro-alpha-D arabinofuranosyl)uracil and the analogous thymidine nucleoside. Compound 6a could not be transformed into nucleosides. PMID- 10795819 TI - Regioselective synthesis of p-nitrophenyl glycosides of beta-D-galactopyranosyl disaccharides by transglycosylation with beta-D-galactosidases. AB - The beta-D-galactosidase from porcine liver induced regiospecific transglycosylation of beta-D-galactose from beta-D-Gal-OC6H4NO2-o to OH-6 of, respectively, p-nitrophenyl glycoside acceptors of Gal, GlcNAc and GalNAc to afford beta-Gal-(1-->6)-alpha-Gal-OC6H4NO2-p, beta-Gal-(1--> 6)-beta-Gal-OC6H4NO2 p, beta-Gal-(1-->6)-alpha-GalNAc-OC6H4NO2-p, beta-Gal-(1-->6)-beta-GalNAc OC6H4NO2-p, beta-Gal-(1-->6)-alpha-GlcNAc-OC6H4NO2-p, and beta-Gal-(1-->6)-beta GlcNAc-OC6H4NO2-p. The enzyme showed much higher transglycosylation activity for the alpha-glycoside acceptors than the corresponding beta-glycoside acceptors. The regioselectivity of the beta-D-galactosidase from Bacillus circulans ATCC 31382 greatly depended on the nature of the acceptor. When alpha-D-GalNAc OC6H4NO2-p and alpha-D-GlcNAc-OC6H4NO2-p were used as acceptors, the enzyme showed high potency for regioselective synthesis of beta-Gal-(1-->3)-alpha-GalNAc OC6H4NO2-p and beta-Gal-(1-->3)-alpha-GlcNAc-OC6H4NO2-p in high respective yields of 75.9 and 79.3% based on the acceptors added. However, replacement of beta-D Gal-OC6H4NO2-p by beta-D-GalNAc-OC6H4NO2-p did change the direction of galactosylation. The enzyme formed regioselectively beta-Gal-(1-->6)-beta-Gal OC6H4NO2-p with (beta-Gal-1-->(6-beta-Gal-1-->)n6-beta-Gal-OC6H4NO2-p, n = 1-4). No beta-(1-->3)-linked product was detected during the reaction. Use of the two readily available beta-D-galactosidases facilitates the preparation of (1-->3)- and (1-->6)-linked disaccharide glycosides of beta-D-Gal-GalNAc and beta-D-Gal GlcNAc. PMID- 10795820 TI - An efficient access to protected disialylated glycohexaosyl threonine present on the leukosialin of activated T-lymphocytes. AB - The total synthesis of the threonine-linked core 2 class disialylated hexasaccharide in a completely protected form was accomplished for the first time. The L-threonine conjugate, N-(9-fluorenylmethoxycarbonyl)-O-[(5-acetamido 4,7,8,9-tetra-O-ben zyl-3,5-dideoxy-D-glycero-alpha-D-galacto-2 nonulopyranosylonic acid)-(2-->3)-(2,6-di-O-benzyl-beta-D-galactopyranosyl)-(1- >4)-2-acetam ido-2-deoxy-3,6-di-O-benzyl-beta-D-glucopyranosyl-(1-->6)-[(5 acetamido- 4,7,8,9-tetra-O-benzyl-3,5-dideoxy-D-glycero-alpha-D-galacto-2-nonulo pyranosylonic acid)-(2-->3)-2,6-di-O-benzyl-beta-D-galactopyranosyl-(1-->3)]-2 acetami do-2-deoxy-alpha-D-galactopyranosyl-(1d-->4c:1f-->4e)-dilactone ]-L threonine allyl ester was synthesized via stereocontrolled glycosylations employing readily accessible monosaccharidic blocks; t-butyl-diphenylsilyl-2 azido-2-deoxy-3,6-di-O-benzyl-beta-D-gluco pyranose, N-(9 fluorenylmethoxycarbonyl)-O-(2-azido-6-O-t-butyldimethylsilyl -2-deoxy-alpha-D galactopyranosyl)-L-threonine allyl ester, 8, 9 and N-(9 fluorenylmethoxycarbonyl)-O-(2-azido-4,6-O-benzylidene-3-O-ch loroacetyl-2-deoxy alpha-D-galactopyranosyl)-L-threonine allyl ester. For the introduction of the amino acid, the azide group was used to temporarily mask the amino group of GalNAc so as to obtain an alpha-glycosidic linkage without participation from the C-2 substituent. The threonine was attached to the sugar unit at the monosaccharide stage to avoid loss of oligosaccharide at a later stage. The Fmoc and allyl ester protected amino acid at the reducing end facilitates efficient glycopeptide synthesis on solid-phase support. PMID- 10795821 TI - Synthesis of 4-cyanophenyl and 4-nitrophenyl 2-azido-2-deoxy-1,5-dithio-beta-D arabino- and -beta-D-lyxopyranosides possessing antithrombotic activity. AB - Tri-O-acetyl-5-thio-D-ribopyranosyl bromide was converted into 3,4-di-O-benzoyl 1,5-anhydro-5-thio-D-erythro-pent-1-enitol (3,4-di-O-benzoyl-5-thio-D-ribal), the azidonitration of which afforded an unstable mixture of 2-azido-3,4-di-O-benzoyl 2-deoxy-1-O-nitro-5-thio-D-pentopyranoside++ + isomers. This was converted without separation into the corresponding 1-O-acetyl derivatives from which an alpha,beta anomeric mixture of the 1-O-acetyl-2-azido-3,4-di-O-benzoyl-2-deoxy-5 thio-D-arabinopyranose+ ++ isomers could be isolated in high yield. Glycosidation of this mixture with 4-cyano- or 4-nitrobenzenethiol, using trimethylsilyl triflate or boron trifluoride etherate, respectively, as promoters gave the corresponding D anomers exclusively. Zemplen debenzoylation afforded 4 cyanophenyl as well as 4-nitrophenyl 2-azido-2-deoxy-1,5-dithio-beta-D arabinopyranoside, respectively. When 1-O-acetyl-2-azido-3,4-di-O-benzoyl-2-deoxy 5-thio-D-lyxopyranose was used as glycosyl donor only the corresponding 1 anomers, i.e., 4-cyanophenyl as well as 4-nitrophenyl 2-azido-2-deoxy-1,5-dithio beta-D-lyxopyranosides, could be isolated after Zemplen debenzoylation in high yield. All four 1,5-dithioglycosides possess significant oral antithrombotic activity. PMID- 10795822 TI - Interconversion of the Ialpha and Ibeta crystalline forms of cellulose by bending. AB - Bending cellulose in a plane normal to the hydrogen-bonded sheets of chains causes a longitudinal displacement of the sheets with respect to one another. The magnitude of this displacement is shown to be sufficient to interconvert the Ialpha and Ibeta forms of cellulose within a bending angle of 39 degrees when the curvature of the sheets of chains comprising the microfibril is modelled as a series of concentric circular arcs. Bending through an angle of 90 degrees is more than sufficient to convert the Ialpha form into Ibeta and back again. Cellulose microfibrils emerging from the cellulose synthase complex in the plasma membrane must bend sharply before they can lie parallel with the inner face of the cell wall. The scale of the changes induced by bending is sufficient to ensure that whatever crystal form would be expected from the geometry of the biosynthetic complex, it is likely be radically altered before the cellulose is incorporated into the cell wall. PMID- 10795823 TI - From amino acid landscape to protein landscape: analysis of genetic codes in terms of fitness landscape. AB - Assigning the values of a certain physicochemical property for individual amino acids to the corresponding codons, we can make an amino acid property "landscape" on a four valued three dimensional sequence space from a genetic code table. Eleven property landscapes made from the standard genetic code (SGC) were analyzed. The evaluation of correlation for each landscape is done by theta value, which represents the ratio of the mean slope (as an additive term) to the degree of roughness (as a nonadditive term). The theta-values for hydropathy indices, polarity, specific heat, and beta-sheet propensity were considerably large with respect to SGC. This implies that the additivity of the contribution from each letter holds for these properties. To clarify the meaning of the so called mutational robustness of SGC, we next examined correlations between the amino acid property and the actual "site fitnesses" of a protein. The site fitnesses were derived from a set of binding preference scores of amino acid residues at every site in MHC class I molecule binding peptides (Udaka et al. in press). We found that the SGC's theta value for an amino acid property is correlated with the significance of the property in the protein function. Adaptive walk simulation on fitness (= affinity) landscapes in a base sequence space for these model peptides confirmed better evolvability due to the introduction of SGC. PMID- 10795824 TI - A phylogenetic perspective on sequence evolution in microsatellite loci. AB - We examined the evolution of the repeat regions of three noncoding microsatellite loci in 58 species of the Polistinae, a subfamily of wasps that diverged over 140 million years ago. A phylogenetic approach allows two new kinds of approaches to studying microsatellite evolution: character mapping and comparative analysis. The basic repeat structure of the loci was highly conserved, but was often punctuated with imperfections that appear to be phylogenetically informative. Repeat numbers evolved more rapidly than other changes in the repeat region. Changes in number of repeats among species seem consistent with the stepwise mutation model, which is based on slippage during replication as the main source of mutations. Changes in repeat numbers can occur even when there are very few tandem repeats but longer repeats, especially perfect repeats led to greater rates of evolutionary change. Species phylogenetically closer to the one from which we identified the loci had longer stretches of uninterrupted repeats and more different motifs, but not longer total repeat regions. The number of perfect repeats increased more often than it decreased. However, there was no evidence that some species have consistently greater numbers of repeats across loci than other species have, once ascertainment bias is eliminated. We also found no evidence for a population size effect posited by one form of the directionality hypothesis. Overall, phylogenetic variation in repeat regions can be explained by adding neutral evolution to what is already known about the mutation process. The life cycle of microsatellites appears to reflect a balance between growth by slippage and degradation by an essentially irreversible accumulation of imperfections. PMID- 10795825 TI - Proteobacterial histidine-biosynthetic pathways are paraphyletic. AB - In Lactococcus lactis there is a protein, HisZ, in the histidine-biosynthetic operon that exhibits significant sequence identity with histidyl-tRNA synthetase (HisRS) but does not aminoacylate tRNA. HisRS homologs that, like HisZ, cannot aminoacylate tRNA are represented in a highly divergent set of bacteria (including an aquificale, cyanobacteria, firmicutes, and proteobacteria), yet are missing from other bacteria, including mycrobacteria and certain proteobacteria. Phylogenetic analysis of the HisRS and HisRS-like family suggests that the HisZ proteins form a monophyletic group that attaches outside the predominant bacterial HisRS clade. These observations are consistent with a model in which the absences of HisZ from bacteria are due to its loss during evolution. It has recently been shown that HisZ from L. lactis binds to the ATP-PRPP transferase (HisG) and that both HisZ and HisG are required for catalyzing the first reaction in histidine biosynthesis. Phylogenetic analysis of HisG sequences shows conclusively that proteobacterial HisG and histidinol dehydrogenase (HisD) sequences are paraphyletic and that the partition of the Proteobacteria associated with the presence/absence of HisZ corresponds to that based on HisG and HisD paraphyly. Our results suggest that horizontal gene transfer played an important role in the evolution of the regulation of histidine biosynthesis. PMID- 10795826 TI - Plastid genome phylogeny and a model of amino acid substitution for proteins encoded by chloroplast DNA. AB - Maximum likelihood (ML) phylogenies based on 9,957 amino acid (AA) sites of 45 proteins encoded in the plastid genomes of Cyanophora, a diatom, a rhodophyte (red algae), a euglenophyte, and five land plants are compared with respect to several properties of the data, including between-site rate variation and aberrant amino acid composition in individual species. Neighbor-joining trees from AA LogDet distances and ML analyses are seen to be congruent when site rate variability was taken into account. Four feasible trees are identified in these analyses, one of which is preferred, and one of which is almost excluded by statistical criteria. A transition probability matrix for the general reversible Markov model of amino acid substitutions is estimated from the data, assuming each of these four trees. In all cases, the tree with diatom and rhodophyte as sister taxa was clearly favored. The new transition matrix based on the best tree, called cpREV, takes into account distinct substitution patterns in plastid encoded proteins and should be useful in future ML inferences using such data. A second rate matrix, called cpREV*, based on a weighted sum of rate matrices from different trees, is also considered. PMID- 10795827 TI - Coevolution of HLA-B and PERB11.1 (MICA): significance of independent triplet expansion within the transmembrane region of PERB11.1 (MICA). AB - Several highly polymorphic sequences are present in the beta block of the MHC, especially HLA-B, HLA-C, PERB11.1 (MICA), and PERB11.2 (MICB). It is now apparent that the polymorphism of PERB11.1 is of the same order as that of HLA-A, -B, and C and it has been suggested that PERB11 could explain some of the disease associations previously attributed to HLA-B. Phylogenetic analysis of PERB11 alpha-domain sequences demonstrates relationships with HLA-B cross-reactive serogroups. In contrast, the transmembrane polymorphisms do not appear to be associated with either PERB11 or HLA-B. These data indicate that PERB11 and HLA-B have evolved in concert from their common ancestors and that the transmembrane polymorphisms have arisen independently and more recently. MHC disease associations will need to be reviewed in the light of mechanisms such as receptor binding and signaling. PMID- 10795829 TI - Implications of fluctuations in substitution rates: impact on the uncertainty of branch lengths and on relative-rate tests. AB - Many tests of the lineage dependence of substitution rates, computations of the error of evolutionary distances, and simulations of molecular evolution assume that the rate of evolution is constant in time within each lineage descended from a common ancestor. However, estimates of the index of dispersion of numbers of mammalian substitutions suggest that the rate has time-dependent variations consistent with a fractal-Gaussian-rate Poisson process, which assumes common descent without assuming rate constancy. While this model does not affect certain relative-rate tests, it substantially increases the uncertainty of branch lengths. Thus, fluctuations in the rate of substitution cannot be neglected in calculations that rely on evolutionary distances, such as the confidence intervals of divergence times and certain phylogenetic reconstructions. The fractal-Gaussian-rate Poisson process is compared and contrasted with previous models of molecular evolution, including other Poisson processes, the fractal renewal process, a Levy-stable process, a fractional-difference process, and a log-Brownian process. The fractal models are more compatible with mammalian data than the nonfractal models considered, and they may also be better supported by Darwinian theory. Although the fractal-Gaussian-rate Poisson process has not been proven to have better agreement with data or theory than the other fractal models, its Gaussian nature simplifies the exploration of its impact on evolutionary distance errors and relative-rate tests. PMID- 10795828 TI - Phylogenetic depth of the bacterial genera Aquifex and Thermotoga inferred from analysis of ribosomal protein, elongation factor, and RNA polymerase subunit sequences. AB - The phylogenetic placement of the Aquifex and Thermotoga lineages has been inferred from (i) the concatenated ribosomal proteins S10, L3, L4, L23, L2, S19, L22, and S3 encoded in the S10 operon (833 aa positions); (ii) the joint sequences of the elongation factors Tu(1alpha) and G(2) coded by the str operon tuf and fus genes (733 aa positions); and (iii) the joint RNA polymerase beta- and beta'-type subunits encoded in the rpoBC operon (1130 aa positions). Phylogenies of r-protein and EF sequences support with moderate (r-proteins) to high statistical confidence (EFs) the placement of the two hyperthermophiles at the base of the bacterial clade in agreement with phylogenies of rRNA sequences. In the more robust EF-based phylogenies, the branching of Aquifex and Thermotoga below the successive bacterial lineages is given at bootstrap proportions of 82% (maximum likelihood; ML) and 85% (maximum parsimony; MP), in contrast to the trees inferred from the separate EF-Tu(1alpha) and EF-G(2) data sets, which lack both resolution and statistical robustness. In the EF analysis MP outperforms ML in discriminating (at the 0.05 level) trees having A. pyrophilus and T. maritima as the most basal lineages from competing alternatives that have (i) mesophiles, or the Thermus genus, as the deepest bacterial radiation and (ii) a monophyletic A. pyrophilus-T. maritima cluster situated at the base of the bacterial clade. RNAP-based phylogenies are equivocal with respect to the Aquifex and Thermotoga placements. The two hyperthermophiles fall basal to all other bacterial phyla when potential artifacts contributed by the compositionally biased and fast evolving Mycoplasma genitalium and Mycoplasma pneumoniae sequences are eschewed. However, the branching order of the phyla is tenuously supported in ML trees inferred by the exhaustive search method and is unresolved in ML trees inferred by the quartet puzzling algorithm. A rooting of the RNA polymerase-subunit tree at the mycoplasma level seen in both the MP trees and the ML trees reconstructed with suboptimal amino acid substitution models is not supported by the EF-based phylogenies which robustly affiliate mycoplasmas with low-G+C gram-positives and, most probably, reflects a "long branch attraction" artifact. PMID- 10795830 TI - Duplication and diversification of the apolipoprotein CI (APOCI) genomic segment in association with retroelements. AB - We have previously shown that several multicopy gene families within the major histocompatibility complex (MHC) arose from a process of segmental duplication. It has also been observed that retroelements play a role in generating diversity within these duplicated segments. The objective of this study was to compare the genomic organization of a gene duplication within another multicopy gene family outside the MHC. Using new continuous genomic sequence encompassing the APOE-CII gene cluster, we show that APOCI and its pseudogene, APOCI', are contained within large duplicated segments which include sequences from the hepatic control region (HCR). Flanking Alu sequences are observed at both ends of the duplicated unit, suggesting a possible role in the integration of these segments. As observed previously within the MHC, the major differences between the segments are the insertion of sequences (approximately 200-1000 bp in length), consisting predominantly of Alu sequences. Ancestral retroelements also contribute to the generation of sequence diversity between the segments, especially within the 3' poly(A) tract of Alu sequences. The exonic and regulatory sequences of the APOCI and HCR loci show limited sequence diversity, with exon 3 being an exception. Finally, the typing of pre- and postduplication Alus from both segments indicates an estimated time of duplication of approximately 37 million years ago (mya), some time prior to the separation of Old and New World monkeys. PMID- 10795831 TI - Coil occlusion of the patent ductus arteriosus: lessons learned. AB - PURPOSE: To review the clinical outcomes of catheter-directed coil occlusion (coil occlusion) of persistently patent ductus arteriosus (PDA) at a pediatric tertiary care hospital. METHODS: A retrospective review of all patients referred to the Cardiac Catheterization Laboratory for coil occlusion at our institution was performed. Twenty-one consecutive patients (12 female) underwent coil occlusion and follow-up between May 1995 and December 1997. We undertook PDA occlusion if: (a) the PDA narrowed to less than 4 mm on echocardiogram and (b) the minimum body weight was approximately 10 kg. Standard right and retrograde left heart catheterization was performed, followed by coil occlusion. Color-flow mapping (CFM) was used intra-procedurally to confirm occlusion of the PDA with a follow-up study several weeks later. RESULTS: The median age and weight of the patients were 33 months and 13.2 kg, respectively. Fourteen patients received one coil, with six requiring a second coil and one requiring multiple coils. Initial follow-up was at a median of 2.4 months. At latest follow-up, 2 patients still have persistent flow at the ductal level. The coils were deployed without complication or embolization. CONCLUSIONS: A review of our first 21 cases demonstrated three important lessons: (1) the maximum diameter of the PDA suitable for coil occlusion is approximately 3 mm; (2) CFM must show complete obliteration of flow in the catheterization lab in order to ensure occlusion of the PDA at follow-up; and (3) the Jackson detachable system allows for precise placement of the coil, often within another coil. PMID- 10795832 TI - A comparison of the efficacy and safety of ioxaglate and iobitridol in renal angioplasty. AB - PURPOSE: To compare ioxaglate and iobitridol for percutaneous transluminal renal angioplasty (PTRA) as regards thromboembolic complications, the quality of diagnosis, and renal and general safety. METHODS: One hundred and eighty-nine patients were prospectively studied, 98 of whom received ioxaglate, and 91, iobitridol. Twenty-two were secondarily excluded from the evaluation of thromboembolic complications as they did not undergo PTRA. RESULTS: Two hundred and two PTRAs were performed. The total volumes of contrast medium administered and the procedure durations were the same for each patient. In the ioxaglate group, four dissections (3 stents), one occlusive dissection, and two spasms occurred; in the iobitridol group, there were three dissections (all stented), one occlusive dissection (stented), and two spasms. The final angiograms showed four renal infarctions with ioxaglate (2 of which were in patients who were not anticoagulated), two with iobitridol. No significant difference was seen in the incidence of thromboembolic complications when the PTRA was performed after anticoagulation (n = 150; 3.9% vs 4%, p = 0.78); in the whole population, thromboembolic complications were more frequent in the ioxaglate group but the difference was not significant (5.7% vs 3.7%, p = 0.74). The quality of the diagnosis and the general and renal safety were the same in the two groups. CONCLUSION: Regarding the clotting phenomenon, we recorded as many thromboembolic complications with ioxaglate as with iobitridol. PMID- 10795833 TI - Primary stenting in infrarenal aortic occlusive disease. AB - PURPOSE: To evaluate the results of primary stenting in aortic occlusive disease. METHODS: Thirty patients underwent primary stenting of focal concentric (n = 2) and complex aortic stenoses (n = 19), and aortic or aorto-iliac occlusions (n = 9). Sixteen patients underwent endovascular outflow procedures, three of whom also had distal open surgical reconstructions. Median follow-up was 16 months (range 1-60 months). RESULTS: Guidewire crossing of two aorto-biiliac occlusions failed, resulting in a 93% (28/30) technical success. Major complications included one access hematoma, one myocardial infarction, one death (recurrent thromboembolism) in a patient with widespread malignancy, and one fatal hemorrhage during thrombolysis of distal emboli from a recanalized occluded iliac artery. One patient did not improve his symptoms, resulting in a 1-month clinical success of 83% (25/30). Following restenting the 26 stented survivors changed their clinical limb status to +3 (n = 17) and +2 (n = 9). During follow-up one symptomatic aortic restenosis occurred and was successfully restented. CONCLUSIONS: Primary stenting of complex aortic stenoses and short occlusions is an attractive alternative to conventional surgery. Larger studies with longer follow-up and stratification of lesion morphology are warranted to define its role relative to balloon angioplasty. Stenting of aorto-biiliac occlusions is feasible but its role relative to bypass grafting remains to be defined. PMID- 10795834 TI - Pleural and pulmonary staining at inferior phrenic arteriography mimicking a tumor staining of hepatocellular carcinoma. AB - PURPOSE: To describe the findings of pleural and pulmonary staining of the inferior phrenic artery, which can be confused with tumor staining during transarterial chemoembolization (TACE) of hepatoma. METHODS: Fifteen patients who showed pleural and pulmonary staining without relationship to hepatic masses at inferior phrenic arteriography were enrolled. The staining was noted at initial TACE (n = 8), at successive TACE (n = 5), and after hepatic surgery (n = 2). The angiographic pattern, the presence of pleural change on computed tomography (CT), and clinical history were evaluated. RESULTS: Draining pulmonary veins were seen in all cases. The lower margin of the staining corresponded to the lower margin of the pleura in 10 patients. CT showed pleural and/or pulmonary abnormalities in all cases. After embolization of the inferior phrenic artery, the accumulation of iodized oil in the lung was noted. CONCLUSION: Understanding the CT and angiographic findings of pleural and pulmonary staining during TACE may help differentiate benign staining from tumor staining. PMID- 10795835 TI - Polyurethane-covered Nitinol Strecker stents as primary palliative treatment of malignant biliary obstruction. AB - PURPOSE: To evaluate the clinical efficacy of the polyurethane-covered Nitinol Strecker stent in the treatment of patients with malignant biliary obstruction. METHODS: Twenty-three covered stents produced by us were placed in 18 patients with malignant biliary obstruction. Jaundice was caused by cholangiocarcinoma (n = 5), pancreatic cancer (n = 6), gallbladder cancer (n = 4), metastatic lymph nodes (n = 2), and tumor of the papilla (n = 1). RESULTS: The mean patency period of the stents was 37.5 weeks (5-106 weeks). Recurrent obstructive jaundice occurred in two patients (11%). Adequate biliary drainage over 50 weeks or until death was achieved in 17 of 18 patients (94.4%). Late cholangitis was observed in two patients whose stents bridged the ampulla of Vater. Other late severe complications were not encountered. CONCLUSION: Although more study is necessary, our results suggest the clinical efficacy of our covered Nitinol Strecker stent in the management of obstructive jaundice caused by malignant diseases. PMID- 10795836 TI - Percutaneous treatment of heterogenous predominantly solid echopattern echinococcal cysts of the liver. AB - PURPOSE: We report our technique for and results of percutaneous treatment of heterogenous, predominantly solid echopattern hepatic hydatid cysts (HHC), i.e., complex type IV cysts according to Gharbi's sonographic classification of HHC. METHODS: Eight patients with nine complex type IV HHC were treated by percutaneous aspiration followed by hypertonic saline ablation, using a 14 Fr van Sonnenberg sump drainage catheter under sonographic and fluoroscopic guidance. RESULTS: Successful drainage of cysts contents was achieved in all eight patients. No major complications, such as anaphylaxis, abdominal dissemination, cyst recurrence, or death, occurred. Minor complications including pain (n = 4), mild fever (n = 5), right reactive pleural effusion (n = 4), and transient hypernatremia (n = 2) were observed and managed conservatively. Follow-up imaging studies for an average period of 15 months (range 1-48 months) showed either complete healing (n = 3) or significant reduction in the size of the cyst with solidification (n = 6). CONCLUSION: Nine complex type IV HHC were effectively treated by suction of the membranes and hypertonic saline ablation using a 14 Fr sump drainage catheter, without major complications. PMID- 10795837 TI - Is lower-dose digital fluorography diagnostically adequate compared with higher dose digital radiography for the diagnosis of fallopian tube stenosis? AB - PURPOSE: In an effort to reduce patient radiation dose during selective fallopian tube catheterization, the diagnostic adequacy of fluoroscopic images was compared with digital radiographic images in both a phantom study and a clinical study. METHODS: For the phantom study polyethylene tubes with inner diameters of 1.30, 0.95, 0.80, 0.57, and 0.45 mm were used. Randomly selected tubes with/without stenoses, recorded by digital radiographic and last-image hold fluoroscopic images, were presented to five blinded radiologists, and receiver-operating characteristic (ROC) analyses were performed. For the clinical study tubal visualization as well as detectability of stenoses and occlusions were analyzed in 14 women using a 2-way analysis of variance for nonrepeated measures. RESULTS: The phantom study showed no significant differences between the two imaging techniques for 0.57-mm-diameter and larger tubes; in contrast, fluoroscopic images provided significantly lower detectability of stenoses in 0.45-mm-diameter tubes (p < 0.05). The clinical study showed inferior tubal visualization and diagnostic performance for fluoroscopic images. CONCLUSIONS: Although fluoroscopic images have inferior diagnostic capability in detection of tubal stenoses and occlusions, these images may be adequate for documenting tubal patency with spill into the peritoneal cavity. PMID- 10795838 TI - Sequestrated thrombolysis: comparative evaluation in vivo. AB - PURPOSE: Lysis of a thrombus is a function of the local concentration of thrombolytic enzymes. This study was designed to determine in a porcine model of acute deep vein thrombosis (DVT) whether perithrombic sequestration of small volumes of a concentrated enzyme solution can accelerate the process of thrombolysis. METHODS: DVT was induced in both hind limbs using a previously described technique (n = 32). Thirty minutes later the animal was heparinized and unilateral thrombolysis was attempted using 8 mg recombinant tissue plasminogen activator (rt-PA); saline was administered in the opposite leg. For conventional high-volume infusion (CI) (n = 5) rt-PA (0.067 mg/ml) was infused at 1 ml/min. For sequestrated thrombolysis the external iliac vein was endoluminally occluded, and rt-PA (0.25 mg/ml) administered either for proximal injection (ST-P) (n = 5), as a bolus every 3 min through a microcatheter placed via the balloon catheter, or for transthrombic injection (ST-T) (n = 5), as a bolus every 3 min through a Katzen wire in the balloon catheter. At autopsy, the thrombus mass in the iliofemoral veins was measured, and the extent of residual thrombosis in the venous tributaries graded at four sites. From these data a thrombolysis score was calculated. RESULTS: One pig died before thrombolysis could be performed. Only with ST-T was residual thrombus mass in the test limb normalized to control, residual thrombus index (RTI), consistently less than unity. The median RTI of this group was 0.50 (range 0.39-0.97) compared with 1.22 (0.64-1.38) for ST-P and 0.88 (0.37-1.13) for CI. Compared with contralateral controls, a lower grade of residual thrombosis in tributaries was observed in test limbs at more venous sites with ST-T (8/20; 95% confidence interval 5-13) and ST-P (9/20; confidence interval 5-13) than with CI (2/20; confidence interval 0-5) (p = 0.04). A trend toward lower thrombolysis scores was observed with ST-T (p = 0.08). Systemic fibrinogenolysis was not observed in any of the groups. Changes in coagulation parameters during thrombolysis were similar irrespective of treatment protocol. CONCLUSIONS: "Transthrombic" sequestrated thrombolysis may offer some advantages over conventional selective infusion for the treatment of acute DVT. However further refinements will be necessary before it can be considered an alternative to the latter. PMID- 10795839 TI - Transhepatic insertion of a metallic stent for the relief of malignant afferent loop obstruction. AB - A 65-year-old man with a polya gastrectomy presented with biliary obstruction. Percutaneous cholangiography indicated strictures of the distal common bile duct and afferent duodenal loop due to an inoperable carcinoma of the head of the pancreas. The patient was unfit for bypass surgery, and a previous gastrectomy precluded endoscopic intervention. Successful palliation of the biliary obstruction was achieved by placing metallic stents across the duodenal and biliary strictures via the transhepatic route. The use of stents for gastrointestinal stricture is reviewed. PMID- 10795840 TI - Iatrogenic pseudoaneurysm in the upper arm: treatment by transcatheter embolization. AB - A 32-year-old woman presented with a pulsatile, painful mass in her left upper arm, originating several days after removal of an Ilizarov external fixation. The diagnosis of a pseudoaneurysm was made by medical history and by physical and ultrasonographic examination of the mass. Angiography confirmed the presence of the pseudoaneurysm, originating from a branch of the arteria profunda brachii, and definitive treatment was performed by transcatheter embolization. Clinical follow-up showed absence of pulsation and pain in the upper arm and a gradual volume decrease of the mass lesion. PMID- 10795841 TI - Diagnosis of saphenous coronary graft aneurysm by spiral computed tomography. AB - We report two cases of coronary saphenous vein graft aneurysms diagnosed by spiral computed tomography. PMID- 10795842 TI - Transjugular intrahepatic portosystemic shunt in a patient with cavernomatous portal vein occlusion. AB - A 23-year-old woman with liver cirrhosis secondary to primary sclerosing cholangitis was referred to us for the treatment of recurrent bleeding from esophageal varices that had been refractory to endoscopic sclerotherapy. Her portal vein was occluded, associated with cavernous transformation. A transjugular intrahepatic portosystemic shunt (TIPS) was performed after a preprocedural three-dimensional computed tomographic angiography evaluation to determine feasibility. The portal vein system was recanalized and portal blood flow increased markedly after TIPS. Esophageal varices disappeared 3 weeks after TIPS. Re-bleeding and hepatic encephalopathy were absent for 3 years after the procedure. We conclude that with adequate preprocedural evaluation, TIPS can be performed safely even in patients with portal vein occlusion associated with cavernous transformation. PMID- 10795843 TI - Ascites due to anastomotic stenosis after liver transplantation using the piggyback technique: treatment with endovascular prosthesis. AB - Liver transplantation preserving the retrohepatic inferior vena cava, the so called piggyback technique, is becoming more frequently used because it avoids caval cross-clamping during the anhepatic phase of surgery. However, hepatic venous outflow blockade causing ascites seems to be less infrequent after piggyback than with cavo-caval anastomosis. We report a 62-year-old patient who underwent liver transplantation using the piggyback technique and developed a stenosis in the anastomosis between the hepatic veins and the inferior vena cava leading to severe postoperative ascites. Ascites was unresponsive to diuretic therapy and was associated with renal function impairment. Since the etiology of the stenosis was mechanical (torsion), percutaneous transluminal angioplasty was unsuccessful. Finally, an autoexpandable prosthesis was placed across the anastomosis resulting in rapid and permanent (3 years of follow-up) resolution of ascites. PMID- 10795845 TI - Malignant superior vena cava obstruction: stent placement via the subclavian route. AB - Over a 3-year period 23 patients with malignant superior vena cava obstruction were referred for interventional management. They underwent repeat localized central venography and deployment of self-expanding Wallstents. All patients (age range 26-89 years) were approached by the subclavian route using 29 stents. The stent was used to exclude thrombus in the contralateral brachiocephalic vein in five patients and histologic information was available in all patients. Retrospective analysis of the clinical records was used to assess symptom-free survival and symptom recurrence. All patients reported an improvement in symptoms within 24 hr of the procedure. There was 100% technical success. Primary clinical success was achieved in 19 of 23 patients followed-up to their death with no symptom recurrence (range 1-34, mean 15 weeks). In four patients symptoms recurred but only one patient was referred for re-intervention, which was successful. Complications included single cases of early post-stent rethrombosis, distal slip on deployment, and distal slip on balloon dilatation. There were no puncture-related complications. PMID- 10795844 TI - Diagnosis of vascular compression at the thoracic outlet using gadolinium enhanced high-resolution ultrafast MR angiography in abduction and adduction. AB - Gadolinium-enhanced magnetic resonance angiography allows rapid evaluation of the vascular structures of the thoracic outlet both in the neutral position and in abduction during one examination within FDA-approved dose limitations for contrast agents. The technique appears to be a good screening one for patients suspected of having vascular thoracic outlet syndrome. PMID- 10795846 TI - Hepatocellular carcinoma extending into the portal vein: restoration of extended term patency by placement of uncovered Wallstents. AB - A 74-year-old man presented with hepatocellular carcinoma extending into the main portal vein. Two bare Wallstents were placed to maintain portal vein patency. The main portal vein remained patent for 6 months after treatment. No serious complications were observed during or after treatment. PMID- 10795847 TI - Polyurethane-covered Wallstents to recanalize Wallstents obstructed by tumor ingrowth from malignant common bile duct obstruction. AB - Four patients with malignant obstruction of the common bile duct had been treated with uncovered Wallstents and suffered from a reobstruction after 2-13 months (mean 5.3 months). Repeat cholangiography revealed severe stenosis of the stent lumen caused by tumor ingrowth through the mesh. A Wallstent with a self-made polyurethane-cover was inserted through the uncovered stent in these patients. The four patients were followed for 3-13 months (mean 6.3 months) until death. There was good drainage with no evidence of recurrent obstruction in all patients. We conclude that a covered Wallstent may extend patency of stented bile ducts, preventing tumor ingrowth in patients with neoplastic obstruction. Further observations are needed. PMID- 10795848 TI - Angioarchitecture of pulmonary arteriovenous malformations: characterization using volume-rendered 3-D CT angiography. PMID- 10795849 TI - Electroconvulsive therapy in adolescence. AB - The aim of this work is to discuss the ethical issues regarding the use of electroconvulsive therapy (ECT) in adolescents. Ethical implications of ECT in adolescents are analyzed in the light of general medical ethics, which include five prominent principles with respect to autonomy, nonmaleficence, beneficence, justice, and cautiousness. As adults, adolescents with acute psychotic impairment raise an inherent conflict between the respect for the patient's autonomy, on the one side, and the principle of beneficence on the other. However, this age group presents particular dilemmas: (i) As any adolescent suffering from a psychiatric illness is a highly vulnerable subject, society asks for particular attention. The consequence of potential overprotection is that the adolescent may remain untreated because of unrealistic fears regarding ECT. (ii) Some of these fears are linked to the cognitive secondary effects of ECT. Although preliminary data are reassuring, more empirical research on this population should be encouraged. (iii) Cautiousness recommends the use of ECT in limited indications catatonia, mood disorders, and intractable acute psychotic disorders. We conclude that there is no ethical reason to ban the use of ECT in adolescents. Ethical options in clinical practice must be evaluated empirically with respect to the consequences for the patient. Dogmatic views should be set aside. PMID- 10795850 TI - Comorbidity and subgroups in childhood autism. AB - The objective of this study was to identify clinical subgroups in children with childhood autism and to examine comorbidity in each subgroup. The study was based on medical records of 30 inpatients referred to a specialist ward for children with pervasive developmental disorders. Eighteen consecutive patients with an IQ above 50 and fulfilling research criteria for childhood autism were grouped in accordance with the Wing's social subgroups of autism (aloof, passive, and active but odd). Comorbidity, in terms of deficits in attention, motor control, visuo motor and visuo-spatial function, as well as epilepsy, was described in each of the three groups. The aloof and passive groups produced a pure autistic triad of deficits, while the active but odd group was characterized by strong comorbidity. PMID- 10795851 TI - Cognitive functions and psychopathological symptoms in early-onset schizophrenia. AB - Type and extent of objectively tested cognitive impairments (attention, verbal fluency, nonverbal reasoning) and their association with self-ratings (Paranoia Depression Scale; Frankfurt Complaint Questionnaire) and clinical assessments (Brief Psychiatric Rating Scale, Scales for the Assessment of Positive Symptoms and Negative Symptoms) of psychopathological symptoms were studied in a sample of 74 adolescents primarily suffering from chronic schizophrenia (DSM-III-R; mean duration of illness = 3.4 years), including 15 patients with a very early onset (< 14 years). Special consideration was given to the differentiation between positive and negative symptoms. In cross-sectional analyses, the schizophrenic adolescents were remarkably impaired in both cognitive functions (attention, reasoning) and psychopathological measures (BPRS, SANS, SAPS). However, factor analysis yielded orthogonal factors for cognitive and psychopathological parameters, and canonical correlation analyses did not find a significant correlation between these two areas. As the degree of objectively measured cognitive impairment in chronic schizophrenic adolescents cannot be predicted by the severity of individual psychopathological symptoms, a multidimensional evaluation of the symptomatology seems to be appropriate. Moreover, premorbid disturbances (motor and/or language developmental disorders) and onset characteristics (age, pattern, subdiagnosis), and their relationship to cognitive impairments were investigated. Premorbid disturbances were confirmed as risk factors for the subsequent occurrence of cognitive impairments. PMID- 10795852 TI - Cerebellar abnormalities in mental illness. A study on Purkinje cell density in schizophrenic men. AB - The linear Purkinje cell density in the cerebellar vermis was investigated in a small cohort of adult onset schizophrenic men with well-documented hippocampal abnormalities. There were no differences in comparison with age-matched controls. This negative finding indicates that vermian abnormalities undoubtedly seen in some schizophrenic patients may constitute a subsyndrome, possibly related to autistic disorders in which cerebellar abnormalities are well corroborated. PMID- 10795853 TI - A natural history of hyperactivity and conduct problems: self-reported outcome. AB - At the age of 16-18 years, outcome was prospectively assessed in a general population sample of four behavioural groups, defined at 6-7 year old: a pure pervasively hyperactive group (N = 31), a mixed hyperactive conduct problem group (N = 20), a pure conduct problem group (N = 18) and a normal control group (N = 29). The objective of the present paper is to describe outcome in those domains for which self-report is recognised as a valid source of information. Differential effects for hyperactivity and conduct problems on outcome were studied. It was found that early hyperactivity and conduct problems predicted different patterns of conduct problems in adolescence. Drug use in adolescence was not predicted by either type of behavioural problem in childhood. Overall social adjustment was worse in the hyperactive groups, whereas no differences in self-esteem were found. Hyperactivity was a strong predictor of relationships problems in adolescence. The results suggest that hyperactivity and conduct problems in childhood are differential predictors of outcome in adolescence. PMID- 10795854 TI - Personality in adolescents with chronic fatigue syndrome. AB - Our aim was to study the presence of personality traits and disorder in adolescents with Chronic Fatigue Syndrome (CFS). Personality was then compared to other measures of functioning such as presence of psychiatric disorder and rating on the Child Behavior Checklist 4-18 (CBCL) and in relation to CFS outcome. Twenty-five adolescents with CFS followed-up after contacts with tertiary paediatric/psychiatric clinics were compared with 15 matched healthy controls. Interviews and questionnaires from parents and youngsters included Personality Assessment Schedule (PAS), Kiddie-SADS Psychiatric Interview, Child Behavior Checklist. CFS subjects were significantly more likely than controls to have personality difficulty or disorder. Personality features significantly more common amongst them were conscientiousness, vulnerability, worthlessness and emotional lability. There was a nonsignificant association between personality disorder and worse CFS outcome. Personality difficulty or disorder was significantly associated with psychological symptoms and decreased social competence on the CBCL but it was distinguishable from episodic psychiatric disorder. Personality difficulty and disorder are increased in adolescents with a history of CFS. Personality disorder may be linked to poor CFS outcome. PMID- 10795855 TI - A 16-year follow-up of a child inpatient population. AB - The purpose of this study was to evaluate long-term outcomes of children who had been inpatients in a child psychiatry unit. Fifty children, who were consecutive admissions to a Dublin inpatient child psychiatry unit between November 1978 and July 1980, were followed up 16 years later. We obtained some direct information on 33 subjects; of these, 4 subjects had died and 11 subjects (33%) were living abroad. A poor outcome as defined by death, imprisonment, adult psychiatric disorder or unemployment was present in 18 of the 24 subjects (79%) on whom this information was available. We conclude that more support and increased availability of services post discharge are needed. PMID- 10795856 TI - Child Behavior Checklist item scores in Norwegian children. AB - In an epidemiological study of children aged four to 16, 1170 parents responded to the Child Behavior Checklist (CBCL). Similar effects of age and socio-economic status (SES) on item scores as those reported in several international studies with the CBCL were found in the total sample. Contrary to the results of other studies, differences related to gender were found for only a few items. A subgroup of 41 children and 36 adolescents and their parents were interviewed with psychiatric interviews. Several CBCL problem items were good predictors of psychiatric disorder in the interviewed children and adolescents. High positive and negative predictive values were found for the items clumsy and the item unhappy, sad or depressed in this sample. PMID- 10795857 TI - Did Ludwig Wittgenstein have Asperger's syndrome? AB - It has been suggested that Ludwig Wittgenstein had Asperger's syndrome. This paper reviews the evidence and concludes that he had the major features of Asperger's syndrome. PMID- 10795858 TI - Only horses and fools. AB - Narrative format, in the form of an invented short story, is used to convey several essential truths in a didactic essay in Neuropsychiatry. Messages, in memorable form, emerge at several levels. Narrative format is essential in biographical medicine and it could be attractive to students at various levels of learning. There are great values and some dangers in biographical science. PMID- 10795859 TI - Adolescent perspectives on the provision of services for their mental health needs. PMID- 10795860 TI - ESCAP Declaration of Hamburg. Research in child and adolescent mental health. PMID- 10795861 TI - Early diagnosis of postoperative infection: assessment of whole blood chemiluminescence. AB - Whole blood chemiluminescence (CL), which allows for the evaluation of both the oxygen radical release by phagocytes and the serum opsonin activity using very small amounts of blood, is considered to be a useful means of assessing the host defense against infection. We measured the whole blood chemiluminescence in 59 patients before and after open surgery (surgery by a laparotomy) for gastrointestinal disease. Early postoperative infection was detected in 12 (20.3%) of the 59 patients. These 12 patients all had significantly higher white blood cell counts compared with the noninfected patients on the third and subsequent days after surgery (P < 0.01). The peak CL in the early postoperative infected group was also significantly higher than that in the noninfected group on the day of surgery (P < 0.0001), 1 day after surgery (P < 0.0001), and 3 days after surgery (P < 0.01). Whole blood CL may therefore be a useful modality for the early detection of postoperative infection in the future. PMID- 10795862 TI - Intrahepatic lithiasis: a Western experience. AB - Intrahepatic lithiasis (IHL) is a very rare disease in Europe and no gold standard treatment has yet been codified. The aim of our study is to report our personal experience of IHL detected in 20 patients (6 males [30%] and 14 females [70%]) between January 1982 and December 1991. Eleven (55%) of these cases presented with only IHL, 7 cases (35%) had IHL associated with gallbladder or common bile duct stones, while 2 (10%) were affected by secondary IHL caused by previous biliodigestive anastomosis-induced stricture. The diagnosis was determined by cholangiography in all patients, by ultrasound in 94.7%, and by computed tomography in 70%. The surgical procedures used were as follows: liver resection in 12 patients (60%); various types of biliodigestive anastomosis in 6 cases (30%); choledocholithotomy with a simultaneous removal of stones from the hepatic duct in 2 patients (10%). The intraoperative mortality rate was nil. Complications consisted of 2 subphrenic abscesses, 1 septicemia, and 1 bowel occlusion. Two patients (10%) dropped out of the follow-up, 2 (10%) died 2 years after surgery for reasons not connected with the disease, 1 (5%) died from suppurative cholangitis 6 months after surgery, 2 (10%) had recurrent stones and were treated by extracorporeal lithotripsy and endoscopic stone removal, while the remaining 13 patients (65%) are symptom-free at follow-up which varied between 6 months and 11 years. Liver resection is the treatment of choice when IHL is confined to one lobe of the liver. When IHL affects the entire liver, a resection of the main involved area should be performed whenever possible. Other options include hepatoduodenal anastomosis using the interposed jejunal loop to enable endoscopic or combined treatment of recurrent stones. PMID- 10795863 TI - Expression of CD44, vascular endothelial growth factor, and proliferating cell nuclear antigen in severe venous invasional colorectal cancer and its relationship to liver metastasis. AB - The first step in liver metastasis is venous invasion by cancer cells from the primary tumor. However, even among cases where the histology shows extensive venous invasion by the primary tumor, we sometimes find cases without synchronous liver metastases. As a result, there is a strong possibility that, besides the established causes of colorectal cancer and that of cancer cells invading the veins, some other important causes for liver metastasis must exist. We investigated the expression rates of CD44, proliferating cell nuclear antigen (PCNA), and vascular endothelial growth factor (VEGF) in 28 primary colorectal tumors using immunohistological techniques, and examined an association with liver metastasis. Cases that are strongly positive for CD44 or PCNA have a higher rate of synchronous liver metastases than cases with either no expression or a low expression. We could find no correlation between the VEGF expression and synchronous liver metastasis. In cases with severe venous invasion, VEGF is not correlated with liver metastasis whereas CD44 and PCNA are correlated with liver metastasis. In cases where severe venous invasion is histologically observed, an immunohistochemical analysis for CD44 and PCNA should be done to assess the likelihood of liver metastases. PMID- 10795864 TI - Predictors of improvement in FEV1 (forced expiratory volume in 1s) after lung volume reduction surgery. AB - We evaluated various preoperative and operative factors to identify the predictors of improvement in forced expiratory volume in 1s (FEV1) after lung volume reduction surgery (LVRS). Fifty-eight emphysema patients received bilateral LVRS either via a sternotomy (n = 53) or by thoracoscopy (n = 5). The patients were divided into the following two groups: group I (n = 17), patients whose FEV1 improved by less than 20%; group II (n = 41), patients whose FEV1 improved by more than or equal to 20%. The preoperative factors (age, degree of dyspnea, oxygen use, steroid use, pulmonary function test, arterial blood gas, pulmonary hemodynamics, 6-min walking distance) and operative factors (removed lung weight, number of staplers) were both similar between the two groups. Upper lobe type emphysema was more frequently seen in group II and the average improvement in FEV1 was significantly better in the patients with upper lobe type emphysema (62.8% +/- 8.5%) than lower lobe type (36.9% +/- 7.6%) and mixed type (35.5% +/- 6.5%), P < 0.01. Severe pleural adhesion was more frequently found in group I and the average improvement in FEV1 was also significantly lower in patients with severe pleural adhesion (19.8% +/- 6.4%) than mild-to-moderate adhesion (46.8% +/- 5.3%) and no-to-minimal adhesion (62.3% +/- 10.4%), P < 0.01. These results lead us to conclude that upper lobe type emphysema may thus be a predictor of better FEV1 improvement while severe pleural adhesion is considered to be a predictor of a poorer FEV1 improvement after bilateral LVRS. PMID- 10795865 TI - Recovery of parathyroid function after total thyroidectomy. AB - To prevent postoperative hypoparathyroidism following total thyroidectomy, the parathyroid glands are preserved in situ and/or resected or devascularized parathyroid glands are autotransplanted. We conducted a retrospective investigation utilizing biochemical and specific endocrine assessments to evaluate the difference in recovery of parathyroid function between the two operative methods. A total of 92 patients underwent total thyroidectomy at our hospital during the period between 1990 and 1997. These patients were divided into a preservation group (n = 83), with one or more preserved glands in situ, and an autotransplantation group (n = 9), with only transplanted glands. The level of intact parathyroid hormone (PTH) was completely restored by 1 year postoperatively in 83% (69/83) of the preservation group patients. In the remaining 14 patients (17%), the intact PTH had fallen below detectable levels on postoperative day (POD) 1, then subsequently recovered to 70% of the preoperative levels. Comparatively, in the autotransplantation group, the mean level of intact PTH recovered to only 43% of the preoperative levels. The results of this study suggest that parathyroid glands should be preserved in situ whenever possible, and that when intact PTH levels fall below detectable limits on POD 1, they may never recover to the preoperative levels in those patients. PMID- 10795866 TI - Current techniques of hyperthermic isolated limb perfusion for malignant melanoma. AB - A retrospective study was conducted examining 25 patients with malignant melanoma who were treated by our new protocol for hyperthermic isolated limb perfusion. The characteristics of our techniques include: a lower priming volume of the extracorporeal circuit; a therapeutic temperature range of 40-41 degrees C with 60 min hyperthermic perfusion; a nominal perfusion flow rate of 500 ml/min in the lower limb and 200 ml/min in the upper limb; and combined carboplatin with interferon-beta as the adjuvant chemotherapy drug. In the lower extremity group, the arterial cannula size ranged from 8 to 14 F, while the venous cannula size ranged from 14 to 16 F. In the upper limb group, the arterial cannula size ranged from 6 to 8F and the venous cannula size ranged from 10 to 12F. No patient required any homologous blood transfusion postoperatively. No operative death or major complications occurred during the early postoperative period, confirming the safety of this treatment. Both optimal cannula size selection and maintaining perfusion temperature below 41 degrees C were judged to be important in elimination of vascular and deep tissue injury. PMID- 10795867 TI - Changes in hepatic venous oxygen saturation in hepatic warm ischemia/reperfusion injury in pigs. AB - To clarify the changes that occur in hepatic venous oxygen saturation (ShVO2) during hepatic ischemia/reperfusion (I/R) injury, we examined the relationship between ShVO2, hepatic tissue blood flow (HTBF), and portal vein pressure (PVP) in a warm I/R model using pig livers. Female pigs weighing 18-23 kg were subjected to warm I/R under extracorporeal circulation between the superior mesentric vein and the left jugular vein to avoid portal congestion. The warm ischemic times were 120 min (n = 4), 180 min (n = 14), and 240 min (n = 4). ShVO2, HTBF, and PVP were measured after reperfusion. The survival rates of the pigs 3 days after reperfusion were 100% in the 120-min group, 57% in the 180-min group, and 25% in the 240-min group. In the 180-min group, the ShVO2 was lower in the pigs that died than in those that survived. There was a significant correlation between ShVO2 and both PVP and HTBF after reperfusion. Histological examination revealed findings of severe I/R injury in pigs with a low ShVO2, and mild I/R injury in pigs with a stable ShVO2. These observations suggest that the changes in ShVO2 could reflect the degree of hepatic I/R injury, especially that related to microcirculatory disturbances occurring at the sinusoid levels. PMID- 10795868 TI - Limitations of exogenous L-arginine in exerting a cytoprotective effect on hepatic ischemia/reperfusion injury. AB - To test whether or not the L-arginine/nitric oxide (NO) pathway induces a protective effect, we investigated the effect of exogenous L-arginine on hepatic ischemia/reperfusion (I/R) injury, using ex vivo perfusion of the isolated rat liver. The rat liver was removed and preserved in cold saline for 60 min, followed by 120 min of reperfusion with oxygenated perfusate at 37 degrees C. Either 600 mg/kg of L-arginine (groups 1 and 4), D-arginine (group 2), N(G)-nitro L-arginine methyl ester (L-NAME) (group 3), or saline (group 5) were administered through the portal vein starting from 5 min before reperfusion to 5 min after reperfusion. In group 4, 600 mg/kg of L-NAME was preadministered at 10 min prior to the administration of L-arginine. The intrahepatic nitric oxide (NO) levels showed only a temporal elevation (227% +/- 70% of the pre-reperfusion levels at 5 min) after reperfusion in group 1. Pretreatment with L-NAME suppressed the elevation of the NO levels immediately after reperfusion in group 4. The lactate dehydrogenase release to the effluent perfusate significantly decreased and the histological findings showed that the sinusoidal damage observed after reperfusion was mitigated in group 1 more than in the other groups. These results thus suggest that exogenous L-arginine produced a relatively small amount of NO and therefore resulted in a slight decrease of hepatic I/R injury. PMID- 10795869 TI - An experimental study of atrial natriuretic peptide levels and the effect of inhaled nitric oxide after pneumonectomy. AB - The aim of this study was to investigate the changes in atrial natriuretic peptide (ANP) levels and lung cyclic guanosine 3',5'-monophosphate (cGMP) concentrations caused by pneumonectomy (Pn), and the effect of inhaled nitric oxide (NO) after Pn in a canine model. The mean pulmonary arterial pressure (PAP) and plasma ANP levels were measured over 180 min in two groups of dogs, one subjected to 60 min of 5 ppm NO inhalation (Pn + NO group, n = 5) and one subjected to 180 min without NO inhalation (Pn group, n = 5). The ANP and cGMP levels in the lung were also measured before and after Pn. Both the PAP and ANP levels increased significantly. Inhaled NO rapidly reduced the PAP and plasma ANP to levels similar to those before Pn. The lung ANP level was significantly increased after Pn, but inhaled NO reduced it to a level similar to that before Pn. The lung cGMP level, which was significantly decreased after Pn, was significantly increased by NO inhalation. These results indicate that NO administration may be effective for preventing post-Pn pulmonary hypertension, although an elevation in ANP does not reduce the PAP. PMID- 10795870 TI - Composite tumor with papillary adenocarcinoma and squamous cell carcinoma of the esophagus: report of a case. AB - Papillary adenocarcinoma is extremely rare in the squamous epithelium-lined esophagus. The histopathologic and immunohistochemical characteristics were examined in a composite tumor showing distinct papillary adenocarcinoma and squamous cell carcinoma of the esophagus resected from a 66-year-old man. The esophageal tumor consisted both grossly and histologically of two distinct components: an ulcerative part showing a squamous cell carcinoma, and a polypoid part corresponding to a papillary adenocarcinoma. In addition, the in situ squamous cell carcinoma was contiguous with the esophageal tumor. Mucin secretion was found only in the papillary adenocarcinoma component. Immunohistochemically, tumor cells of the papillary adenocarcinoma component were positive for carcinoembryonic antigen, secretory component, and lactoferrin. These staining patterns were similar to those of the normal esophageal gland proper. These histologic, mucin-histochemical, and immunohistochemical findings suggest that the papillary adenocarcinoma originated from the submucosal esophageal gland and the squamous cell carcinoma from the squamous epithelium lining the esophagus. PMID- 10795871 TI - Metachronous triple cancers of the sigmoid colon, stomach, and esophagus: report of a case. AB - We report herein an unusual case of metachronous triple cancers of the sigmoid colon, stomach, and esophagus. A 60-year-old man was initially admitted to our hospital for investigation of occult fecal blood. This was found to be caused by sigmoid colon cancer which was resected in July 1985 (T3, N0, M0; Stage II). A follow-up endoscopy performed in 1990 showed early gastric cancer, and a gastrectomy was performed in August 1990 (Tis, N0, M0; Stage 0). Another endoscopic examination performed as follow-up in 1993 revealed early cancer of the remnant stomach, and all the remnant stomach was surgically resected in March 1993 (Tis, N0, M0; Stage 0). He presented again in December 1996, complaining of discomfort in the chest which was found to be caused by cancer of the middle thoracic esophagus. Although surgery was considered necessary, the patient refused to undergo any further operations. Instead, radiation was administered from January 1997. An endoscopy after the completion of radiotherapy confirmed that the cancer had almost disappeared; however, it started to grow again from the beginning of 1998. He was hospitalized due to esophageal stenosis in April 1998, and died of carcinomatous cachexia in September of the same year. PMID- 10795872 TI - Rapid growth of a retroperitoneal rhabdomyosarcoma following right hemicolectomy for ascending colon cancer: report of a case. AB - We report herein the case of a 69-year-old man in whom rapid growth of a retroperitoneal rhabdomyosarcoma occurred following hemicolectomy for ascending colon cancer. On his first admission for surgery, a small lesion, 1.5 cm in diameter, was detected adjacent to the inner side of the left kidney by abdominal axial computed tomography (CT), which was initially suspected to be a benign lesion; however, a postoperative follow-up CT scan done 5 months later revealed that the lesion had enlarged remarkably to 8 cm in diameter. Thus, total resection was performed under the presumed diagnosis of a malignant retroperitoneal tumor. The tumor was found to be adjacent to the inner portion of the left kidney and covered by Gerota's fascia. As it involved the ileolumbar muscle and had a metastatic lymph node, complete resection was performed. The resected specimen was 8.5 x 6.5 x 5 cm in size and was histologically confirmed as a retroperitoneal rhabdomyosarcoma of embryonal type. Two courses of adjuvant chemotherapy with adriamycin, vincristine, and cyclophosphamide were given, and the patient has shown no signs of recurrence for 2 years since his second operation. PMID- 10795873 TI - Primary lymphoma of the liver: report of a case. AB - We describe herein a female patient with non-Hodgkin's lymphoma of the liver and present a review of the related literature. The patient was referred with the diagnosis of malignant hemangiopericytoma (with an open biopsy). The physical examination, standard laboratory test results and tumor marker levels were all normal. A nonstandard left lobectomy was performed. Histopathological and immunohistochemical examinations revealed non-Hodgkin's lymphoma of B-cell type. The findings of a peripheral blood smear and bone marrow biopsy were normal. There was no other site of involvement based on physical or radiological examinations. These findings established the diagnosis of primary hepatic lymphoma. Fewer than 100 cases have been reported in the world literature. The best treatment results have been obtained by a resection followed by chemotherapy when feasible. PMID- 10795874 TI - Pancreatic lesions in von Hippel-Lindau syndrome: the coexistence of metastatic tumors from renal cell carcinoma and multiple cysts. AB - Multiple cysts and benign cystadenomas of the pancreas have been documented occasionally in von Hippel-Lindau syndrome (HLS); however, the malignant involvement of the pancreas in HLS is very rare. We report a case of HLS in which metastatic tumors from renal cell carcinoma (RCC) coexisted with multiple cysts in the pancreas. A 22-year-old woman with a history of HLS had undergone a partial resection of the left kidney for RCC 3 years earlier, at which time a solid mass in the pancreatic tail and multiple pancreatic cysts were also incidentally detected by computed tomography. Over the following 3 years, the mass enlarged slightly, thus raising suspicions that it might be a primary neoplasm of the pancreas. She was referred to the Department of Surgery and Surgical Basic Science to undergo surgery. In addition to the tumor in the pancreatic tail, however, further tumors in the pancreatic head were also disclosed by preoperative celiac arteriography and intraoperative palpation and ultrasonography. A distal pancreatectomy was performed, because the enucleation of all the tumors in the pancreatic head was technically impossible and because the patient declined a total pancreatectomy. A histologic examination of the mass in the pancreatic tail revealed metastatic RCC. This case emphasizes that metastatic disease should be included in the differential diagnosis when evaluating the pancreas in a patient with HLS. PMID- 10795875 TI - Splenic vein occlusion secondary to tuberculous lymphadenitis at the splenic hilum: report of a case. AB - We report a patient with splenic vein occlusion (SVO) secondary to tuberculosis. A 17-year-old male patient with mild epigastric pain and splenomegaly was found to have gastric varices by gastroscopy, and SVO by selective angiography. At operation, the splenic vein was occluded by hard fibrous tissue at the splenic hilum, and thus a splenectomy was performed. A microscopic examination of the tissue revealed caseous necrosis surrounded by epithelioid cells and Langhans type giant cells. Although there were no other findings suggesting intestinal tuberculosis, it seemed that tuberculous lymphadenitis of the splenic hilum most likely caused the occlusion of the splenic vein. Because specific tests for tuberculosis were negative in both immunohistochemical staining for bacille Calmette-Guerin and polymerase chain reaction of DNA for Mycobacterium tuberculosis, the time of infection was assumed to have occurred a long time before. SVO can sometimes be seen in pancreatic diseases, but this patient with tuberculosis appears to be the first such reported case in the English literature. PMID- 10795876 TI - Congenital hemangiopericytoma: report of a case. AB - We report a case of congenital hemangiopericytoma arising in the lower right leg of a 4-day-old male neonate. Despite the generally good prognosis associated with this neoplasm, a complete surgical excision has so far been recommended to avoid recurrences, because no definite criteria for determining whether or not the tumor will regress spontaneously have been established to date. PMID- 10795877 TI - Total arch replacement with an aortic arch aneurysm due to chronic interstitial pneumonia: report of a case. AB - The case of a 66-year-old man who had rheumatoid arthritis, chronic interstitial pneumonia (IP) with honeycomb lung, and an aortic arch aneurysm is described. He complained of left chest pain in April 1998 and chest computed tomography revealed an enlargement of a thoracic aneurysm whose maximum diameter reached 7 cm. He was urgently transferred to our institution to undergo immediate surgery for an impending rupture of the aneurysm. His PaO2 with 80 Torr with oxygen therapy. The operation included a median sternotomy, extracorporeal circulation with selective cerebral perfusion, the use of cold blood cardioplegic solution, and open distal anastomosis. Although an acute exacerbation of IP occurred 2 months after the surgery, he successfully received intensive care including high dose steroid therapy and artificial ventilation. He was successfully weaned from the ventilator and is now being followed up with medical treatment. PMID- 10795879 TI - Regeneration of lymphatic vessels and lipometabolism in an experimental rat model of extensive dissection of the abdominal periaortic lymphatic vessels and lymph nodes. AB - The present study was conducted to analyze the regeneration of lymphatic vessels and lipometabolism in an experimental rat model of extensive dissection of the abdominal periaortic lymphatic vessels and lymph nodes from the renal artery to the mesenteric and celiac artery. Findings showing the regeneration of mesenteric lymphatic vessels were classified as complete, incomplete, or no regeneration. The incomplete regeneration of fine irregular lymphatic vessels was observed in all rats, 2 and 3 weeks postoperatively. At the same time, triglyceride (TG) was found to be significantly lower (P < 0.05) in the rats that were given extensive dissection than in the control group, due to chylomicron blockade after lymphatic dissection. The TG recovered to within the normal range around 4 weeks postoperatively following the growth of fine regenerated lymphatic vessels or collateral route formation. PMID- 10795878 TI - Involvement of the right atrium by malignant lymphoma as a cause of right cardiac failure: report of a case. AB - We describe herein a rare case of malignant lymphoma occupying almost the entire space of the right atrial cavity and causing low cardiac output syndrome. A life saving emergency operation was carried out after the establishment of a temporary bypass between the axillary and femoral veins to prevent exacerbation of the patient's condition during the induction of anesthesia. Cardiopulmonary bypass was commenced and the right atrium was opened. A large tumor in the right atrium could not be completely removed due to invasion of the atrial wall. A bypass from the left innominate vein to the pulmonary arterial trunk was constructed with a prosthetic graft to convert the blood flow directly from the systemic vein to the pulmonary artery. Postoperative radiation treatment was given, which resulted in reducing the size of the tumor considerably, and the patient is doing well 1 year after his operation. PMID- 10795880 TI - Importance of preoperative marking for minithoracotomy and for internal thoracic artery harvesting in minimally invasive direct coronary artery bypass grafting. AB - Minimally invasive direct coronary artery bypass has the potential to cause an anastomotic failure because of a limited exposure of the operative field and the difficulty of internal thoracic artery harvesting. In the present study, the importance of preoperative marking for an accurate minithoracotomy location and a successful internal thoracic artery harvest was assessed. A paperclip was placed on the left nipple and a chest X-ray was performed in the supine position. By aligning the position of the paperclip to the location of the left anterior descending coronary artery from a coronary arteriogram frontal view, the intercostal space for the minithoracotomy was thus determined. Marking the incisional intercostal space during preoperative left internal thoracic arteriography revealed the number and location of the internal thoracic artery branches at the beginning of the harvest. This preoperative marking technique allowed for a more adequate exposure of the operative field and an easier internal thoracic artery harvest which therefore contributed to an improvement in the operative results. PMID- 10795881 TI - Oxidative stress in amyotrophic lateral sclerosis. AB - The pathogenesis of amyotrophic lateral sclerosis is poorly understood. In one or two percentage of patients, mutations in the SOD1 gene are known to underly the disease. Even in these cases, the mechanism of cell death remains unclear. Most researchers agree that damage by reactive oxygen species is involved in this process, but whether the latter plays a primary role or is an epiphenomenon is uncertain. As evidence for oxidative stress is not only found in mutant SOD1 related familial amyotrophic lateral sclerosis, but also in sporadic amyotrophic lateral sclerosis, it is tempting to speculate that a similar mechanism is at work in both forms of the disease. PMID- 10795882 TI - The role of excitotoxicity in ALS--what is the evidence? AB - It is well accepted that excitotoxic mechanisms contribute to the pathogenesis of acute neuronal death in stroke, epilepsy, or brain trauma. It is less widely acknowledged that excitotoxic mechanisms play a role in the pathogenesis of chronic neurological disorders, in particular neurodegenerative diseases. However, evidence is accumulating that this mechanism is indeed part of the pathogenesis of late-onset neurodegenerative diseases. One of the clinical examples may be amyotrophic lateral sclerosis, a disease in which antiexcitotoxic strategies have neuroprotective effects in both, an established animal model and in man. In addition, there is accumulating neuropathological, pathobiochemical and pathophysiological evidence which indicates that excitotoxic mechanisms are part of the pathogenesis of the human disease and consequently part of the mechanisms explaining selective vulnerability ("pathoclisis") in the human motor system. PMID- 10795884 TI - Motor neurones in culture as a model to study ALS. AB - Defining the basis of the selective cell vulnerability of motor neurones (MN) represents the key issue in amyotrophic lateral sclerosis (ALS), and tissue culture models are the ideal system for the identification of the MN specific features at the single cell level. Neurone-astrocyte metabolic interactions, which have a critical role in MN through glutamatergic toxicity, have been mostly defined in vitro. Ca++ metabolism, which appears to play a critical role in inducing MN loss in ALS, has been successfully studied using in vitro cell models. Furthermore, primary cultures demonstrated that apoptotic or necrotic death of neurones after injury depends upon the cell energetic status. Superoxide dismutase- (SOD-1) mutations were successfully expressed in cultured rodent MNs, providing a critical assay to sequence the molecular processes responsible for MN degeneration due to the identified genetic defect. The recent identification of genes that separate humans from apes further increases the value of the human in vitro models to better understand specific human cellular properties. Purified human MNs and astrocytes can today be obtained from the human embryonic spinal cord anterior horns. Interactions at the single cell level can be dissected using the cDNA amplification techniques. The effects of molecules affecting MN survival, neurite extension, and metabolism can easily be defined in vitro, gaining a critical mass of information of immediate clinical application in the treatment of patients affected by ALS. Understanding the properties of human MNs in vitro represents today a significant and critical tool that can easily be reached after extension of the available knowledge from non-primate to human research. Human MN culture studies can greatly contribute to identifying the primitive critical cellular events responsible for the MN degeneration observed in ALS and to gaining crucial information on new therapeutical agents. PMID- 10795883 TI - Molecular factors underlying selective vulnerability of motor neurons to neurodegeneration in amyotrophic lateral sclerosis. AB - Current research evidence suggests that genetic factors, oxidative stress and glutamatergic toxicity, with damage to critical target proteins and organelles, may be important contributory factors to motor neuron injury in amyotrophic lateral sclerosis (ALS). Various molecular and neurochemical features of human motor neurons may render this cell group differentially vulnerable to such insults. Motor neurons are large cells with long axonal processes which lead to requirements for a high level of mitochondrial activity and a high neurofilament content compared to other neuronal groups. The lack of calcium buffering proteins parvalbumin and calbindin D28k and the low expression of the GluR2 AMPA receptor subunit may render human motor neurons particularly vulnerable to calcium toxicity following glutamate receptor activation. Motor neurons also have a high perisomatic expression of the glutamate transporter protein EAAT2 and a very high expression of the cytosolic free radical scavenging enzyme Cu/Zn superoxide dismutase (SOD1) which may render this cell group vulnerable in the face of genetic or post-translational alterations interfering with the function of these proteins. More detailed characterisation of the molecular features of human motor neurons in the future may allow the strategic development of better neuroprotective therapies for the benefit of patients afflicted by ALS. PMID- 10795885 TI - The role of SMN in spinal muscular atrophy. AB - Childhood spinal muscular atrophy (SMA) is a common autosomal recessive disorder which is characterized by muscle weakness due to degeneration of motoneurons in the spinal cord and brainstem nuclei. Positional cloning strategies have revealed several gene candidates including the genes for the survival motoneuron (SMN) and the neuronal apoptosis inhibitory protein (NAIP). Both genes are duplicated on chromosome 5. Homozygous deletions/mutations of the telomeric SMN gene, which is expressed from both copies on human chromosome 5, are associated with the disease. Recent reports suggest involvement of the SMN protein in the formation of spliceosomal particles in the cytoplasm and in the regeneration of spliceosomes in the nucleus. These data put spinal muscular atrophy into a growing group of disorders of RNA metabolism which also include fragile-X syndrome and myotonic dystrophy. Relevance of these previous data for the pathogenesis of the disease are discussed in this review. PMID- 10795887 TI - Radiation-induced aneurysm and moyamoya vessels presenting with subarachnoid haemorrhage. AB - A rare case of a ruptured internal carotid artery (ICA) aneurysm, with ICA stenosis and moyamoya vessels, was diagnosed in a young female patient after radiotherapy for optic glioma. Retrospective analysis revealed that an extraordinarily high dose of radiation (110 Gy) had been delivered to the area around the ICA in two therapeutic sessions. Clipping and wrapping of the aneurysm were performed and the patient made a good recovery. A review of the literature suggested that rupture of radiation-induced aneurysms is a rare event, but may be fatal. Considerable care must therefore be taken when delivering high-dose radiation to cerebral arteries. We suggest that the incidence of radiation induced moyamoya vessels is higher in younger patients. PMID- 10795886 TI - Growth factors in gliomas revisited. AB - Overexpression or untimely expression of wild-type or mutated protein growth factors and their receptors is associated with the biology of malignant gliomas and other types of cancer. It may result in unchecked tumour cell proliferation, migration/invasion into normal tissue, tumour angiogenesis, escape from immune surveillance, and decreased apoptotic cell death, i.e. after treatment with cytotoxic agents. This often involves activation of growth factor receptors either by simultaneous production of growth factors and corresponding receptors on the same or adjacent tumour cells or by constitutive receptor activation due to mutations. In several instances, the cellular genes encoding these growth factors and receptors are homologous to transforming genes/oncogenes from tumourigenic retroviruses and have thus been regarded as cellular proto oncogenes. In recent years much progress has been made towards a better understanding of the function of these molecules and how they lead to the aggressive phenotype of malignant gliomas and its inherent resistance to adjuvant therapies. This, still insufficient, knowledge is a prerequisite for the development of novel therapies for this non-curable disease. The aim of this review is to address relevant growth factor receptor systems with emphasis on their particular role in glioma biology. PMID- 10795888 TI - Cerebral arteriovenous malformations in children. AB - In spite of their congenital origin, only 18-20% of cerebral AVMs are diagnosed during infancy and childhood. Intracranial haemorrhage is the presenting clinical manifestation in 75-80% of paediatric patients and is associated with a high morbidity and mortality. The natural history of untreated cerebral AVMs in children is worse than in adults, in relation to a longer life expectation, a higher annual risk of AVM bleeding (3.2% vs. 2.2%) and a higher incidence of posterior fossa and basal ganglia AVMs, most of which present with massive haemorrhages. The surgical excision remains the treatment of choice for parenchymal AVMs in children; AVM complete removal is currently achieved in 70 90% of the patients. With the advent of new agents for endovascular management, preoperative AVM embolization has further improved surgical results. Stereotactic radiosurgery appears to be a successful treatment option in small or moderate sized AVMs. Recent studies have demonstrated low complication rates with this technique in paediatric patients. We reviewed our experience with 37 paediatric AVMs treated at the Section of Paediatric Neurosurgery of the Catholic University of Rome between 1980 and 1997. Twenty-three patients underwent surgery as the only treatment modality; endovascular embolization was combined with the surgical treatment in a further four cases. Radiosurgery was utilized as the only treatment in three patients and in combination with other techniques in an other three children (with surgery in one case and with AVM embolization in the remaining two subjects). No treatment was carried out in three patients because of excessively critical condition on admission; endovascular embolization failed in a further patient because of the anatomical complexity of the malformation. Previous studies have demonstrated a quite strict correlation between AVM complexity based on Spetzler and Martin's grading system and patients outcome. A less direct relationship has been observed in the present study. In our experience the factors which were more closely predictive of patients' outcome were the occurrence of an AVM bleeding and the neurological status on admission. In spite of the low number of cases in the single subsets of patients this study seems to support the role of AVM embolization and radiosurgery as effective adjuvant techniques in the management of cerebral AVMs in children. PMID- 10795889 TI - Clinical safety and performance of Sugita titanium aneurysm clips. AB - BACKGROUND: In spite of advantages for neuro-imaging, titanium aneurysm clips are not yet chosen for routine use for clipping of intracranial aneurysms, probably because their reliability has not been demonstrated in large numbers of cases. We assess reliability and safety of Sugita titanium aneurysm clips for routine clinical use in a large number of operated cases. METHOD: Intra-operative performance and safety of Sugita titanium aneurysm clips were evaluated in 347 patients in our institutions. Aneurysms in 261 patients had ruptured: 86 patients had unruptured aneurysms. RESULTS: A total of 441 clips of 52 different types were used. No early or delayed complications occurred in relation to the titianium clips, such as deformity or slippage of a clip. Patient outcome according to the Glasgow Outcome Scale was similar to that when conventional cobalt-based aneurysm clips were used. INTERPRETATION: Reliability and safety of Sugita aneurysm clips were demonstrated in a large number of patients. The clips are suitable for routine use in aneurysm surgery. PMID- 10795890 TI - Meningioma manifesting intracerebral haemorrhage: a possible mechanism of haemorrhage. AB - We present a possible mechanism of intracerebral peritumoural haemorrhage in meningioma based on the clinical data of three of our cases. A meningioma manifesting intracerebral haemorrhage is uncommon and some sporadic case reports have been presented, but without any proven mechanisms. We are presenting three cases of convexity meningioma manifesting spontaneous intracerebral haemorrhage with apoplectiform onset. All three patients had no evidence of bleeding tendency or other predisposing factors for haemorrhage. Preoperative radiological studies showed a solid mass attached to the dura with intracerebral peritumoural haematoma. Total removal of the tumour and haematoma could be achieved in every case. Histological investigation revealed extensive tumour infarction in two cases and fibrosis related to pre-existing ischaemia in the other case. The diagnoses were atypical meningioma in two cases and transitional type in one case. We suggest that extensive tumour infarction might be a cause of spontaneous intracerebral peritumoural haemorrhage in our series of patients. PMID- 10795891 TI - Posteroventral pallidotomy in Parkinson's disease. AB - OBJECTIVE: We analyse the clinical aspects, results and reliability of posteroventral pallidotomy (PVP) carried out as treatment for the principal symptoms and treatment induced complications in patients with Parkinson's disease (PD). PATIENTS AND METHODS: Between August 1995 January 1998, 17 patients with PD were treated surgically, 13 patients with PVP. A pre- and post-surgical clinical evaluation was carried out. Riechert's Stereotactic System (MHT, Freiburg, Germany) was used. Ventriculography under stereotactic conditions was used in the PVP procedures, Laitinen's co-ordinates as anatomical target, and electrical stimulation for physiological determination. 3 4 radiofrequency lesions were made at 83 degrees C for 20". The mean age was 60 +/- 10.8 years, ages ranging from 45 79 years. 8 (60.5%) of the patients were male. The cardinal symptoms of the series were bradykinesia and rigidity. The duration of the illness ranged from 8.6 +/- 3.7 years with a range of 4-15 years. 7 (53.8%) presented with a duration of 10 years or more. 6 (46,2%) of the patients underwent left PVP, the remaining 7 (53.8%) right PVP. Only one patient received treatment with right PVP and ipsilateral thalamotomy in the same surgical procedure. The mean post-surgical follow up was 16 +/- 7 months, with a range of 2 to 26 months. RESULTS: An up to date evaluation was carried out on all patients showing significant changes after PVP in UPDRS motor (P < 0.005), complete rigidity relief (P < 0.005), bradykinesia relief (P < 0.005) and complete tremor relief (P < 0.005). An important improvement in contralateral dyskinesia was noted after PVP. A subjective evaluation of the results showed excellent results in 4 (30.8%) patients, good in 6 (46.2%) and fair in 3 (23%). No significant correlation was found between age and duration of illness (P = 0.7). Two patients suffered slight side effects, one patient with worsening of hypophonia whilst the other suffered subjective visual impairment controlled by normal post operative ophthalmological examinations. There was no peri-operative mortality. CONCLUSION: PVP is considered a safe and effective surgical method for the treatment of both the principal symptoms of PD and the complications of DOPA medication. PMID- 10795892 TI - Open MRI-guided microsurgery of intracranial tumours. Preliminary experience using a vertical open MRI-scanner. AB - OBJECTIVES: A number of different image-guided surgical techniques have been developed during the past decade. None of these methods can provide the surgeon with information about the dynamic changes that occur intra-operatively. The development of open configurated MRI-scanners leads to new perspectives in the intra-operative management and resection control of intracranial tumours. MATERIAL AND METHOD: Using a vertical open 0.5 T MRI-scanner for intra-operative MR image guided neurosurgery, forty-four patients (20 female/24 male) with different intracranial tumours have been operated on since August 1997. The patients ranged in age from 20 to 70 years (mean +/- standard deviation = 47.2 +/ 15.9 ys). RESULTS: In 36 (82%) of 44 patients the tumours were completely removed with the aid of MR image-guidance. In 8 cases (18%) complete removal was not achieved. Postoperatively 6 (14%) of 44 patients developed neurological deficits which were transient in 5 cases (paresis, dysphasia). In these patients the tumours were located in or near eloquent brain areas (sensorimotor cortex/speech center). CONCLUSION: Intra-operative MRI is helpful for navigation as well as determining of tumour margins to achieve a complete and safe resection of intracranial lesions. Complications related to the surgical procedure are reduced and the risk of neurological deterioration due to tumour removal and postoperative complications is minimized. It can be concluded that the intra operative application of interventional MRI technology may represent a major step foreward in the field of neurosurgery. PMID- 10795893 TI - Early experience with poly L-lactic acid bioabsorbable fixation system for paediatric craniosynostosis surgery. Report of 3 cases. AB - OBJECTIVE: The authors describe early experience in the use of bioabsorbable fixation devices for cranial reconstruction of paediatric craniosynostosis patients. METHODS: Three patients, individually respectively presenting sagittal synostosis, metopic synostosis, and clover leaf skull deformity, underwent cranial reconstruction using poly L-lactic acid (PLLA) plates and screws. The patients ranged in age from 2 to 10 months at the time of surgery. The postoperative clinical follow-up ranged from 16 to 18 months. All patients showed satisfactory wound healing without signs of infection or local inflammation. The plates provided satisfactory fixation and were not palpable through the skin after 16 to 18 postoperative months. CONCLUSION: Our experience demonstrated the efficacy of PLLA bioabsorbable plates and screws for cranial reconstruction in cases of infant craniosynostosis. Prospective studies and longer follow-up of a larger number of patients is desirable for confirmation of these findings. PMID- 10795894 TI - Delayed administration of the K+ channel activator cromakalim attenuates cerebral vasospasm after experimental subarachnoid hemorrhage. AB - BACKGROUND: Delayed cerebral vasospasm remains an unpredictable and inadequately treated complication of aneurysmal subarachnoid hemorrhage (SAH). Recent evidence indicates that the potassium channel activator cromakalim is capable of limiting cerebral vasospasm in rabbits when administered immediately after experimental SAH (i.e. before spastic constriction has been initiated). However, the ultimate clinical value of cromakalim for treating vasospasm will depend in part on its effectiveness when administered after SAH-induced constriction has already been initiated. The present study examined the effects of cromakalim on vasospasm when treatment was initiated after SAH-induced constriction was underway. METHODS: New Zealand white rabbits were subjected to experimental SAH by injecting autologous blood into the cisterna magna. Cromakalim (0.03, 0.1 or 0.3 mg/kg) or vehicle was injected intravenously at 8 hour intervals beginning 24 hours post-SAH. Animals were killed by perfusion fixation 48 hours after SAH. Basilar arteries were removed and sectioned, and cross-sectional area was measured. FINDINGS: The average cross sectional areas of basilar arteries were reduced by 64% and 68% in the SAH-only and SAH + vehicle groups, respectively. Treatment with cromakalim dose-dependently attenuated SAH-induced constriction. The groups treated with 0.03, 0. 1, and 0.3 mg/kg cromakalim exhibited average decreases in cross sectional area of 57%, 42%, and 19%, respectively. INTERPRETATION: These findings indicate that cromakalim dose-dependently attenuates cerebral vasospasm when administered 24 hours after experimental SAH in the rabbit. The results suggest K(ATP) channel activators, such as cromakalim. could be of benefit for reversing cerebral vasospasm after aneurysmal SAH. PMID- 10795895 TI - S-100B protein serum levels after controlled cortical impact injury in the rat. AB - S-100B is described to provide information about the severity of brain damage in man. Estimation of serum markers appears to be an easy method of obtaining information regarding severity and outcome after head injury. However less is known about the post traumatic time course of this protein in the serum. The aim of this study was to provide further information about the posttraumatic enzymekinetik. 65 male Wistar rats were subjected to severe cortical impact injury (100 PSI, 2 mm deformation). Blood samples were drawn directly after trauma, then after 1 h, 6 h, 12 h, 24 h, and 48 h. In sham operated animals blood samples were drawn directly after craniotomy, then after 6 h and after 48 h. Also compared were S-100B serum levels at different severities in 20 rats (45 PSI, 75 PSI; 2 mm deformity) after controlled cortical impact to sham operated animals. S 100B serum levels were estimated with a commercially available enzyme immuno assay (DAKO). The mean serum level in the sham group was 0.38 microg/l. Serum levels at 100 PSI differed statistically significantly directly after trauma up to 24 h. The 48 h S-100B levels showed no significant difference in the sham group. Serum levels at different severities differed significantly from the sham group, but did not differ concerning level of severity. The controlled cortical impact model is able to produce a raised serum level of the S-100B protein for 24 hours. Different trauma severities were not reflected. PMID- 10795896 TI - Subarachnoid haemorrhage as initial symptom of multiple brain abscesses. AB - The case of evolving multiple brain abscesses which became symptomatic with a sudden hemianopsia and the clinical and radiological signs of a subarachnoid haemorrhage, is reported. A common pathomechanism which could explain both the sudden focal neurological deficit and the subarachnoid bleeding is discussed. PMID- 10795897 TI - A case of subcortical meningioma. AB - Subcortical meningioma, which has been reported as meningioma without dural attachment, a cerebral subcortical lesion, is extremely rare. Very few findings of radiological examination of subcortical meningioma have been described. Pre operative differentiation of this lesion is generally difficult. We characterize the peritum oural oedema adjacent to the meningioma in this region, and we suggest that radiological findings of peritum oural oedema contribute to differential diagnosis of subcortical meningiomas. PMID- 10795898 TI - Eosinophilic granuloma of the orbit in an adult woman. PMID- 10795899 TI - Partial recovery of paraplegia due to spontaneous intramedullary ependyma haemorrhage. PMID- 10795900 TI - Duplication of the middle cerebral artery associated with an unruptured aneurysm. PMID- 10795901 TI - Veni, vidi, vici: the neurohypophysis in the twentieth century. AB - We outline the key discoveries in the first 70 years of research on the neurohypophysis that provided the foundations for more recent studies in the last 30 years. We consider the extent to which these recent studies, which have exploited molecular technologies, cellular electrophysiological techniques and mechanistic behavioural investigations, have advanced or changed our understanding of the functions of oxytocin and vasopressin. The different evolutionary pressures on the oxytocin and vasopressin systems are discussed. Lastly, we focus on the mechanisms underlying the burst-firing activity of oxytocin neurones in lactation as a problem not yet solved, and probably requiring a presently improbable conceptual leap to understand. PMID- 10795902 TI - Discovery and design of novel and selective vasopressin and oxytocin agonists and antagonists: the role of bioassays. AB - Synthetic oxytocin and vasopressin agonists and antagonists have become important tools for research and were instrumental in the identification of the four known receptor subtypes, V1a, V2, V1b (V3) and oxytocin, of these peptide hormones. However, the relative lack of receptor selectivity, particularly of the antagonists, has limited their usefulness as experimental probes and their potential as therapeutic agents. We now present some findings from our continuing studies aimed at the design of more selective oxytocin and vasopressin agonists and antagonists and a structure-activity relationship update on our recently discovered novel hypotensive vasopressin peptides. Bioassays have been, and continue to be, of critical importance in leading to the discovery of the novel agonists, antagonists and hypotensive peptides reported here. This paper highlights three main aspects of these studies. (1) Replacement of the tyrosine2 and/or phenylalanine3 residues in the V2 agonist deamino,[Val4,D-Arg8]arginine vasopressin (dVDAVP) by thienylalanine resulted in selective V2 agonists with strikingly high potencies. However, the peptide solutions were unstable and lost activity over time. These highly potent V2 agonists, which are devoid of vasopressor activity, are promising leads for improving drugs for treating diabetes insipidus, enuresis and coagulation disorders. (2) Diaminopropionic acid and diaminobutyric acid substitution at position-5 in oxytocin and in V1a antagonists yielded, respectively, the first specific antagonist for the oxytocin receptor, desGly-NH2,d(CH2)5[D-Trp2,Thr4,Dap5]OVT and the first specific antagonist for the vasopressin V1a receptor, d(CH2)5[Tyr(Me)2,Dab5]AVP. The availability of single receptor subtype-specific or selective antagonists will enhance our ability to delineate receptor functions. Utilising these new receptor specific probes, we were able to show that the uterotonic action of vasopressin is mediated principally by oxytocin and not by V1a receptors. (3) Replacement of the phenylalanine3 residue in the V1a/V2/oxytocin antagonist, d(CH2)5[D Tyr(Et)2,Val4]AVP, with arginine3 yielded the novel, selective, hypotensive vasopressin peptide, d(CH2)5[D-Tyr(Et)2,Arg3,Val4]AVP (Peptide I). Bioassay characterisations of Peptide I show that its vasodepressor action is independent of the peripheral autonomic, bradykinin, nitric oxide and prostaglandin systems and is not mediated by the known classical oxytocin and vasopressin receptors. These findings suggest the existence of a new vasopressin receptor subtype that may be relevant to the vasodilating action of vasopressin in regional vascular beds. Iodinatable hypotensive peptides have been synthesised and could be developed as markers for the putative new receptor. Ongoing structure-activity relationship studies on Peptide I have led to more potent and selective hypotensive peptides for use as new research tools and as leads for the development of a new class of antihypertensive agents. PMID- 10795903 TI - Regulation of vasopressin V1b receptors in the anterior pituitary gland of the rat. AB - Vasopressin secreted by parvocellular neurones of the hypothalamic paraventricular nucleus modulates pituitary adrenocorticotrophic hormone (ACTH) secretion by acting upon vasopressin V1b type receptors in the pituitary corticotroph coupled to phospholipase C. Regulation of V1b receptors contributes to the adaptation of the hypothalamic-pituitary-adrenal (HPA) axis to stress, as evidenced by the correlation between vasopressin receptor number and pituitary ACTH responsiveness. V1b receptor upregulation during chronic stress is associated with elevated circulating glucocorticoids and vasopressin expression in parvocellular neurones, suggesting that these factors control V1b receptor expression. Removal of circulating glucocorticoids by adrenalectomy causes sustained vasopressin receptor downregulation, but reduces V1b receptor mRNA only transiently. The latter effect is not mediated by increased corticotrophin releasing hormone (CRH) and vasopressin release, since it is not prevented by lesions of the hypothalamic paraventricular nucleus. Adrenalectomy causes sustained V1b receptor loss in Brattleboro rats, which lack hypothalamic vasopressin, suggesting that vasopressin mediates V1b receptor mRNA recovery. Exogenous glucocorticoid administration downregulates pituitary vasopressin binding but increases V1b receptor mRNA and facilitates coupling of the receptor to phospholipase C, effects which may contribute to the refractoriness of vasopressin actions to glucocorticoid feedback. The lack of parallelism between changes in pituitary vasopressin binding and V1b receptor mRNA levels during manipulation of the HPA axis indicates that V1b receptor content depends on post transcriptional mechanisms rather than steady-state V1b receptor mRNA levels. These studies suggest that interaction between glucocorticoids and vasopressin plays an important role in regulating V1b receptor mRNA expression during alterations of the HPA axis. In addition, the recent characterization of a major part of the V1b receptor gene provides a basis for studying the molecular mechanisms regulating the V1b receptor. PMID- 10795904 TI - Gene regulation of vasopressin and vasopressin receptors in cancer. AB - It is proposed that neuropeptide production by tumours is an important part of a special process of oncogenic transformation rather than a pre-existing condition of progenitor cells; this concept is called Selective Tumour gene Expression of Peptides essential for Survival (STEPS). All small-cell lung cancers and breast cancers evidently express the vasopressin gene, and this gene seems to be structurally normal in all but exceptional cases. Vasopressin gene expression in cancer cells leads to the production of both normal and abnormal forms of tumour vasopressin mRNA and proteins. Although the necessary post-translational processing enzymes are expressed in these cells, most processing seems to be extragranular, and most of the protein products become components of the plasma membrane. Small-cell lung cancer and breast cancer cells also express normal genes for all vasopressin receptors and produce normal vasopressin receptor mRNAs and V1a and V1b receptor proteins, and the vasopressin-activated calcium mobilising (VACM) protein; plus both normal and abnormal forms of the V2 receptor. Through these receptors, vasopressin exercises multifaceted effects on tumour growth and metabolism. A normal protein vasopressin gene promoter seems to be present in small-cell lung cancer cells, and this promoter contains all of the transcriptional elements known to be involved in gene regulation within hypothalamic neurones. Since these elements largely account for regulation of tumour gene expression observed in vitro, it is likely that as yet unknown factors are selectively produced by tumours in vivo to account for the observed seemingly autonomous or unregulated production of hormone in tumour patients. Promoter elements thought to be responsible for selective vasopressin gene expression in small-cell lung cancer probably include an E-box and a neurone restrictive silencer element close to the transcription start site. It is possible that transcription factors acting at these same elements can explain selective vasopressin expression, not only in small-cell tumours, but also in all other tumours such as breast cancer. By extrapolation, similar mechanisms might also be responsible for the expression of additional features that characterize the 'neuroendocrine' profile of these cancers. PMID- 10795905 TI - Oxytocin receptors and cholesterol: interaction and regulation. AB - Cholesterol affects the ligand binding function of the oxytocin receptor in a highly specific manner. While the structurally-related cholecystokinin receptor shows a strong correlation between the membrane fluidity and its binding function, the oxytocin receptor behaves differently. A stringent and unique requirement of the affinity state of the oxytocin receptor for structural features of the sterol molecule has been found. The molecular requirements differ both from those postulated for sterol-phospholipid interactions and from those known to be necessary for the activity of other proteins. Employing a new detergent-free subcellular fractionation protocol, a two-fold enrichment of the oxytocin receptors (10-15% of total receptors) has been detected in the cholesterol-rich, caveolin-containing membrane domains of the plasma membrane. While most of the properties of the oxytocin receptors were indistinguishable in cholesterol-poor versus cholesterol-rich membrane compartments, high-affinity oxytocin receptors localised in caveolin-enriched low-density membranes showed about a 3-fold higher stability against thermal denaturation at 37 degrees C compared with the oxytocin receptors localised in high-density membranes. Moreover, addition of cholesterol to the cholesterol-poor high-density membranes fully protected the oxytocin receptors against thermal denaturation and partially rescued high-affinity oxytocin binding. Although the membrane fluidity of the caveolin-enriched domains was lower than that in the high-density membranes, there was no correlation between the stability of oxytocin receptors and the fluidity level of the membrane domains. Finally, in a molecular modelling approach a putative cholesterol binding motif on the extracellular surface of the oxytocin receptor was found. PMID- 10795906 TI - Signal pathways mediating oxytocin stimulation of prostaglandin synthesis in select target cells. AB - A major action of oxytocin is to stimulate prostaglandin production in reproductive tissues. The two major enzyme systems involved are cytosolic phospholipase A2 (cPLA2), which catalyses the formation of arachidonic acid from membrane glycerophospholipids, and prostaglandin endoperoxide-H synthases-1 and 2, which allow conversion of arachidonic acid to prostaglandins. During gestation, the concentrations of all three enzymes rise in the rabbit amnion. Agonists, including oxytocin, increase cPLA2 activity, in part, by elevating intracellular Ca2+ concentration, which causes cPLA2 to be translocated from the cytosol to intracellular membrane binding sites. Cytosolic PLA2 is then activated by a mitogen-activated protein kinase (MAPK)-dependent step. Our studies have elucidated signal pathways involved in oxytocin-stimulated prostaglandin output in both rabbit amnion cells and Chinese hamster ovary cells stably transfected with the rat oxytocin receptor. The two cell types are alike with respect to oxytocin-stimulated intracellular Ca2+ transients, mediation via Gq, and the specific MAPK that catalyses the phosphorylation of cPLA2. However, they differ with respect to the mechanisms of upregulation of key enzymes involved in prostaglandin E2 synthesis. These findings illustrate the tiers of complementary mechanisms involved in oxytocin stimulation of prostaglandin E2, and the extent of the diversity in the cellular signalling pathways involved. PMID- 10795907 TI - Molecular basis of ligand binding and receptor activation in the oxytocin and vasopressin receptor family. AB - Although it is now widely accepted that G-protein-coupled receptors exist in at least two allosteric states, inactive and active, and that the spontaneous equilibrium between the two is regulated by various events including the binding of specific agonists and antagonists, the molecular counterparts of these functionally different states are still poorly understood. In this paper, we review our current knowledge concerning the structure-function relationships of the oxytocin and vasopressin receptors, focusing in particular on the process of receptor activation. Using a combined approach of site-directed mutagenesis and molecular modelling, we investigated the molecular events leading to agonist dependent and -independent receptor activation in the human oxytocin receptor. Our analysis allows us to propose that the active conformations of this receptor are characterised by similar rearrangements of its cytosolic regions that ultimately lead to the opening of a putative docking site for the G-protein. Furthermore, the dynamics of these motions are similar to that observed in the alpha1B-adrenergic receptor, thus suggesting that, although activated by different ligands, the process of receptor isomerization in these two receptors is regulated by the same cluster of highly conserved residues and that common molecular events are responsible for receptor activation in different G-protein coupled receptors. PMID- 10795908 TI - Excitatory versus inhibitory modulation by ATP of neurohypophysial terminal activity in the rat. AB - Much is now known about the electrophysiological properties of the magnocellular neurones of the hypothalamus. Oxytocin neurones are characterized by an intermittent high frequency discharge during suckling that leads to the pulsatile release of oxytocin into the blood and to subsequent milk ejection. Vasopressin neurones are characterized by their asynchronous phasic activity (bursting) during maintained vasopressin release and the subsequent regulation of water balance. In both cases, it is the clustering of spikes, albeit with different time courses for each peptide, that facilitates hormone release. The mechanism underlying this differential facilitation is one of the major unanswered questions in neuroendocrinology. This paper considers recent evidence that indicates that ATP, co-secreted with vasopressin and oxytocin, may play a key role in the regulation of stimulus-secretion coupling in the neurohypophysis. The activity of the type (II) Ca2+-activated K+ (K(Ca)) channel found in the nerve terminals was significantly increased in the presence of ATP on the cytoplasmic side of the channel. Extracellular ATP, in contrast, inhibited the type II K(Ca) current in a dose-dependent manner. Thus, intracellular and extracellular ATP exert opposite effects on the type II K(Ca) channel of neurohypophysial terminals. Furthermore, ATP opens P2X2 channels to increase intracellular [Ca2+] in the nerve terminals and subsequent arginine vasopressin (AVP) release. In contrast, adenosine, acting via A1 receptors, specifically inhibits only the N type Ca2+ channel, thus decreasing neuropeptide release. These multiple, conflicting effects of ATP and its metabolite adenosine could explain the patterns of AVP release observed during physiological stimulation in vivo. PMID- 10795909 TI - Intracellular calcium signalling in magnocellular neurones of the rat supraoptic nucleus: understanding the autoregulatory mechanisms. AB - Oxytocin and vasopressin, released at the soma and dendrites of neurones, bind to specific autoreceptors and induce an increase in [Ca2+]i. In oxytocin cells, the increase results from a mobilisation of Ca2+ from intracellular stores, whereas in vasopressin cells, it results mainly from an influx of Ca2+ through voltage dependent channels. The response to vasopressin is coupled to phospholipase C and adenylyl-cyclase pathways which are activated by V1 (V1a and V1b)- and V2-type receptors respectively. Measurements of [Ca2+]i in response to V1a and V2 agonists and antagonists suggest the functional expression of these two types of receptors in vasopressin neurones. The intracellular mechanisms involved are similar to those observed for the action of the pituitary adenylyl-cyclase activating peptide (PACAP). Isolated vasopressin neurones exhibit spontaneous [Ca2+]i oscillations and these are synchronised with phasic bursts of electrical activity. Vasopressin modulates these spontaneous [Ca2+]i oscillations in a manner that depends on the initial state of the neurone, and such varied effects of vasopressin may be related to those observed on the electrical activity of vasopressin neurones in vivo. PMID- 10795910 TI - Adolescent stress and neural plasticity in hamsters: a vasopressin-serotonin model of inappropriate aggressive behaviour. AB - Animal studies show that arginine vasopressin facilitates aggression, while serotonin (5-HT) inhibits aggression by blocking the activity of the vasopressin system. Clinical studies report that subjects with a history of 'fighting and assault' show a significant positive correlation between cerebrospinal fluid concentrations of vasopressin and aggression in the presence of a hyporeactive 5 HT system. Thus, in animals and humans, a hyporeactive 5-HT system may result in enhanced vasopressin activity and increased aggression. Can the stress of emotional and physical insult, i.e. threat and attack, during adolescence affect the development of the vasopressin and 5-HT systems and alter normal aggressive behaviour in early adulthood? Adolescent male golden hamsters were weaned at postnatal day 25, and stressed for 2 weeks by daily 1 h bouts of threat and attack by adult hamsters. Male littermates were run in a parallel stress study using daily 1 h trials of isolation in a novel environment. During early adulthood, on postnatal day 45, 3 days after the cessation of stress trials, animals were tested for aggression in a resident: intruder model. The results show a context-dependent change in aggression. Animals with a history of abuse show exaggerated attack behaviour toward smaller males compared to littermates with a history of isolation stress. Conversely, when confronted by males of equal size, animals with a history of abuse show diminished aggression and increased submission compared to controls. It was determined that the density of vasopressin fibres and neurones in the hypothalamus is lower in abused animals compared to controls. In contrast, the number of 5-HT terminals within the hypothalamus is higher in abused animals compared to controls. These results provide evidence in an animal model that stress in the form of threat and attack during adolescence can alter the balance between vasopressin and 5-HT in the brain, resulting in inappropriate aggressive behaviour in early adulthood. PMID- 10795911 TI - The role of oxytocin and regulation of uterine oxytocin receptors in pregnant marsupials. AB - The oxytocin-like peptide of most Australian marsupials is mesotocin, which differs from oxytocin by a single amino acid. This substitution has no functional significance as both peptides have equivalent affinity for and biological activity on the marsupial oxytocin-like receptor. A role for mesotocin in marsupial parturition has been demonstrated in the tammar wallaby where plasma mesotocin concentrations increase less than one minute before birth. Infusion of an oxytocin receptor antagonist at the end of gestation disrupts normal parturition, probably by preventing mesotocin from stimulating uterine contractions. In the absence of mesotocin receptor activation, a peripartum surge in prostaglandins is delayed which suggests a functional relationship between mesotocin, prostaglandin release and luteolysis. Female marsupials have anatomically separate uteri and in monovular species, such as the tammar wallaby, only one uterus is gravid with a single fetus whereas the contralateral uterus remains non-gravid. We have used this unique animal model to differentiate systemic and fetal-specific factors in the regulation of uterine function during pregnancy. The gravid uterus in the tammar wallaby becomes increasingly sensitive to mesotocin as gestation proceeds, with the maximum contractile response observed at term. This is reflected in a large increase in mesotocin receptor concentrations in the gravid uterus, and a downregulation in the non-gravid uterus in late pregnancy. The upregulation in myometrial mesotocin receptors is pregnancy-specific and independent of systemic steroids. One factor that may influence mesotocin receptor upregulation in the gravid uterus in late pregnancy is mechanical stretch of the uterus caused by the growing fetus. Our data highlight that a local fetal influence is more important than systemic factors in the regulation of mesotocin receptors in the tammar wallaby. PMID- 10795912 TI - Catecholaminergic mechanisms underlying neurohypophysial hormone responses to unconditioned or conditioned aversive stimuli in rats. AB - Oxytocin release from the neurohypophysis is facilitated by systemic cholecystokinin octapeptide (CCK) administration and noxious stimuli. Oxytocin release after CCK administration is mediated by A2 noradrenergic neurones while the release after noxious stimuli appears to be mediated by A1 noradrenergic neurones. On the other hand, facilitation of vasopressin release after noxious stimuli is not dependent upon noradrenergic neurones but on dopamine receptors. Environmental stimuli previously paired with noxious stimuli (conditioned fear stimuli) or novel environmental stimuli facilitate oxytocin release and suppress vasopressin release. These neuroendocrine responses to conditioned fear stimuli, but not to novel stimuli, are impaired by central noradrenaline depletion or i.c.v. adrenoceptor antagonists. These data suggest that there are at least two types of stress responses in neuroendocrine systems, one noradrenaline dependent, and one noradrenaline independent. It is also suggested that noradrenergic neurones are functionally heterogeneous in the control of oxytocin release. PMID- 10795913 TI - Oxytocin, motherhood and bonding. AB - Release of the peptide hormone oxytocin in the brain has been shown to influence both maternal, sexual and social bonding behaviours although there are a number of species differences. This review summarizes findings on the distributions of oxytocin and oxytocin receptors in the brain, together with factors governing their expression, release of the peptide in the brain and its behavioural actions. A model of how oxytocin may act to alter maternal and socio-sexual behaviours is proposed which initially involves activation of oxytocin neurones in a single brain site, the paraventricular nucleus of the hypothalamus (PVN), following vaginal and cervical stimulation. This causes a co-ordinated release of the peptide in the PVN and its terminal projection regions for up to 1 h and this promotes different behavioural components, primarily through modulation of classical transmitter systems. PMID- 10795914 TI - Behavioural impact of intraseptally released vasopressin and oxytocin in rats. AB - The two nonapeptides arginine vasopressin and oxytocin are not only secreted from the neurohypophysis into the general circulation but are also released intracerebrally. Our recent research has focused on the release patterns and effects of oxytocin and vasopressin in brain areas, such as the septum and hypothalamus, that are thought to be involved in the regulation of (1) behavioural responses and (2) responses of the hypothalamo-neurohypophysial system (HNS) to stressor exposure in rats. The results demonstrate that combined physical and emotional stress (induced by exposure to forced swimming) selectively triggers the release of vasopressin within all brain areas under study but not into the general circulation. Under emotional stress conditions (induced by exposure to the 'social defeat' procedure), however, oxytocin rather than vasopressin release increased within the hypothalamus and septum. Experiments aimed at revealing the neuroendocrine and behavioural relevance of the local nonapeptide release provided evidence for an involvement of vasopressin in the regulation of HNS activity (within the hypothalamus) and, moreover, in acute stress-coping strategies, anxiety-related behaviour and learning and memory processes (within the septum). The observed dissociation between central and peripheral nonapeptide release not only supports the hypothesis that plasma vasopressin and oxytocin concentrations do not necessarily reflect central release patterns but also suggests vasopressin and oxytocin neurones are able to independently release their nonapeptide from different parts of their neuronal surface (e.g. from somata/dendrites vs. axon terminals). This remarkable regulatory capacity provides the basis for an differential involvement of vasopressin, and probably also oxytocin, in the co-ordination of neuroendocrine activity, emotionality and cognition at different brain levels to ensure an appropriate behavioural response of the organism to stressful stimuli. PMID- 10795915 TI - Dendritic secretion of peptides from hypothalamic magnocellular neurosecretory neurones: a local dynamic control system and its functions. AB - The role of the dendrites of magnocellular neurones in the release of neurosecretory peptides and the synthesis of many proteins locally is reviewed. Oxytocin and vasopressin contained in dense-cored neurosecretory vesicles are released from magnocellular dendrites not only by excitatory transmitters such as glutamate acting through well-established receptors, but also by a rapid action of oestradiol acting by a mechanism which appears to involve NMDA receptors. Magnocellular dendrites also contain substantial amounts of the synthetic machinery which could synthesise proteins for local use. The presence in dendrites of polysomes and of mRNAs encoding microtubule-associated protein 2, calcium calmodulin kinase II, alpha-synapsin-associated protein, and components of the GABA(A) and NMDA receptors strongly suggests that these proteins can be translated in the dendrites, close to the sites at which they function. Mechanism(s) which control the translation of these dendritic mRNAs and the insertion into the dendritic membranes of proteins translated by dendritic ribosomes remain to be determined. However, an overall picture emerges of magnocellular dendrites as active secretory and synthetic components of the neurosecretory neurones. PMID- 10795916 TI - Neurohypophysial peptides as retrograde transmitters in the supraoptic nucleus of the rat. AB - A possible role for vasopressin and oxytocin in the physiology of the supraoptic nucleus was investigated using nystatin-perforated patch recording in acute brain slices from the rat containing the supraoptic nucleus. We observed that exogenously applied oxytocin reduced glutamate-mediated synaptic transmission by acting at a presynaptic oxytocin receptor. Endogenous oxytocin, released either by afferent excitation (tetanus) or by postsynaptic depolarization of the recorded magnocellular neurone caused a similar reduction of excitatory input and this could be blocked with an oxytocin antagonist. Thus endogenous oxytocin, released from magnocellular dendrites, acts as a retrograde transmitter to reduce afferent excitation. We discuss the possible significance of these results in terms of the physiological role of oxytocin in the intact animal and suggest possible avenues for further experimentation. PMID- 10795917 TI - Pre- and postsynaptic modulation of the electrical activity of rat supraoptic neurones. AB - The release of vasopressin and oxytocin is regulated by the electrical activity of magnocellular neurosecretory cells in the supraoptic and paraventricular nuclei, which is under the control of a great variety of neurotransmitters and neuromodulators. The major neural signals to the supraoptic nucleus are from excitatory glutamate inputs and inhibitory GABA inputs. In recent studies, the voltage-clamp mode of the whole-cell patch-clamp technique has been applied to slice preparations from rat hypothalamus to monitor synaptic inputs to supraoptic neurones. Spontaneous excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) are abolished by CNQX and picrotoxin, respectively, but are insensitive to tetrodotoxin, indicating that they represent quantal release of glutamate and GABA, respectively, from nerve terminals of presynaptic neurones. GABA and glutamate show remarkable suppressive effects on both EPSCs and IPSCs via presynaptic GABA(B) and mGlu receptors, respectively. Noradrenaline, which excites supraoptic neurones via postsynaptic alpha1-receptors, also suppresses IPSCs and potentiates EPSCs. On the other hand, prostaglandin E2, which excites supraoptic neurones via postsynaptic prostaglandin E2 (EP) receptors of the EP4 subclass, also suppresses IPSCs via EP3 receptors but has little effect on EPSCs. Thus pre- and postsynaptic mechanisms may act cooperatively to excite supraoptic neurones. Nitric oxide, which inhibits supraoptic neurones, potentiates IPSCs without affecting EPSCs. This provides another example for the preferential modulation of IPSCs of supraoptic neurones. On the other hand, PACAP, which causes a long-lasting increase in the firing frequency via the postsynaptic receptors, has no effect on EPSCs and IPSCs, suggesting that some ligands act only at postsynaptic receptors. Thus multiple patterns for pre- and postsynaptic modulation are present in the supraoptic nucleus, and the electrical activity of supraoptic neurones is regulated via complex mechanisms at both pre- and postsynaptic sites. PMID- 10795918 TI - Local circuitry regulates the excitability of rat neurohypophysial neurones. AB - The importance of angiotensin II (AII) and glutamate has long since been recognized in neuroendocrine regulation. However, the mechanisms by which AII and glutamate modulate the excitability of the paraventricular nucleus (PVN) have largely remained a mystery until recently. It is now apparent that AII and glutamate are potent stimulators of both magnocellular and parvocellular neurones in the rat PVN. While glutamate, the predominant excitatory neurotransmitter in the CNS, ubiquitously excites PVN neurones, AII appears to mediate excitability of the PVN by both direct and indirect mechanisms. Interestingly, both of these neurotransmitters, upon exciting the PVN, activate an inhibitory feedback system, which is capable of diminishing the initial stimulus. Physiologically, this moderates the output signals from the PVN, and probably also regulates neuropeptide release from the neurohypophysis. The importance of this negative feedback loop is evident in the pathophysiological implications of a disruption in the system. Evidence suggests that a breakdown in this system may be responsible in part for the onset and maintenance of both congestive heart failure and hypertension. Future studies will continue to characterize both the actions of glutamate and AII in the PVN, and to further elucidate the mechanisms which control the excitability of the PVN. PMID- 10795919 TI - A physiological role for adrenomedullin in rats; a potent hypotensive peptide in the hypothalamo-neurohypophysial system. AB - Adrenomedullin, a potent hypotensive peptide, was originally isolated from human phaeochromocytoma. Adrenomedullin immunoreactivity and gene expression are found not only in peripheral organs but also in the central nervous system. Adrenomedullin labelled cells were localised in the hypothalamus, including in the paraventricular and supraoptic nuclei, in rats. Abundant adrenomedullin immunoreactive fibres and varicosities were found in the hypothalamo neurohypophysial tract and the internal zone of the median eminence in colchicine treated and hypophysectomized rats, whereas in control rats few adrenomedullin labelled fibres were observed. We examined the effects of intracerebroventricular administration of adrenomedullin on neurosecretory cells in the paraventricular and supraoptic nuclei of rats, using immunohistochemistry for Fos protein and in situ hybridisation histochemistry for c-fos mRNA. Intracerebroventricular administration of adrenomedullin caused a marked induction of Fos-like immunoreactivity in the paraventricular nucleus and the dorsal part of the supraoptic nucleus. In the paraventricular and supraoptic nuclei, nuclear Fos like immunoreactivity was predominantly in oxytocin-immunoreactive cells rather than vasopressin-immunoreactive cells. The induction of c-fos mRNA in the paraventricular and supraoptic nuclei was increased in a dose-related manner 30 min after intracerebroventricular administration of adrenomedullin. This induction was reduced by pre-treatment with the adrenomedullin receptor antagonist, human adrenomedullin-(22-52)-NH2. Intracerebroventricular administration of adrenomedullin also caused a marked increase in the plasma concentration of oxytocin. Extracellular recordings from magnocellular neurosecretory cells in the paraventricular nucleus revealed that putative oxytocin-secreting cells were activated by intracerebroventricular administration of adrenomedullin. These results suggest that central adrenomedullin preferentially stimulates the secretion of oxytocin by activating hypothalamic oxytocin-secreting cells and may have an important role in salt appetite and body fluid homeostasis in rats. PMID- 10795920 TI - The role of steroid hormones in the regulation of vasopressin and oxytocin release and mRNA expression in hypothalamo-neurohypophysial explants from the rat. AB - Vasopressin and oxytocin release from the neural lobe, and the vasopressin and oxytocin mRNA contents of the supraoptic and paraventricular nuclei are increased by hypertonicity of the extracellular fluid. The factors regulating these parameters can be conveniently studied in perifused explants of the hypothalamo neurohypophysial system that include the supraoptic nucleus (but not the paraventricular nucleus) with its axonal projections to the neural lobe. Vasopressin and oxytocin release and the mRNA content of these explants respond appropriately to increases in the osmolality of the perifusate. This requires synaptic input from the region of the organum vasculosum of the lamina terminalis. Glutamate is a likely candidate for transmitting osmotic information from the organum vasculosum of the lamina terminalis to the magnocellular neurones, because agonists for excitatory amino acid receptors stimulate vasopressin and oxytocin release, and because increased vasopressin release and mRNA content induced in hypothalamo-neurohypophysial explants by a ramp increase in osmolality are blocked by antagonists of both NMDA (N-methyl-D-aspartate) and non-NMDA glutamate receptors. Osmotically stimulated vasopressin release is also blocked by testosterone, dihydrotestosterone, oestradiol and corticosterone. Both oestrogen and dihydrotestosterone block NMDA stimulation of vasopressin release, and in preliminary studies oestradiol blocked AMPA stimulation of vasopressin release. Thus, steroid inhibition of osmotically stimulated vasopressin secretion may reflect inhibition of mechanisms mediated by excitatory amino acids. Recent studies have demonstrated numerous mechanisms by which steroid hormones may impact upon neuronal function. Therefore, additional work is warranted to understand these effects of the steroid hormones on vasopressin and oxytocin secretion and to elucidate the potential contribution of these mechanisms to regulation of hormone release in vivo. PMID- 10795921 TI - Diurnal rhythms in neurohypophysial function. AB - The neurohypophysial hormones oxytocin and vasopressin show daily rhythms of secretion with elevated hormone release during the hours of sleep. This pattern can be modulated by ovarian steroids and alters with age. The pattern appears to be due in part to the nocturnal increase in melatonin secretion, which stimulates hormone release in man, while being inhibitory in the rat. Pinealectomy alters both the 24 h pattern of neurohypophysial hormone release in the rat and the firing rate of magnocellular supraoptic nucleus neurones. There is also a reduced hormone release in response to hypovolaemia and raised plasma sodium concentration compared to sham operated animals, with a smaller increase in neuronal activity, as determined by immediate-early gene expression. The normal responses can be restored by nocturnal administration of melatonin. Melatonin also influences the neurohypophysial hormone response in the human to known stimuli of release, such as raised plasma osmolality, exercise and insulin induced hypoglycaemia. Recent studies have revealed that not only does the release of vasopressin and oxytocin vary over each 24 h, but the respective renal and pregnant uterine responses also show diurnal variations. PMID- 10795922 TI - Differential expression of two adhesion molecules of the immunoglobulin superfamily, F3 and polysialylated NCAM, in hypothalamic magnocellular neurones capable of plasticity. AB - The adult hypothalamo-neurohypophysial system undergoes activity-dependent, reversible morphological changes which result in reduced astrocytic coverage of its neurones and an increase in their synaptic contacts. Our recent observations show that neurones and glia of the hypothalamo-neurohypophysial system continue to express 'embryonic' molecular features which may underlie their capacity to undergo such plasticity. These include expression of cell surface molecules like the glycosyl phosphatidyl inositol (GPI)-linked glycoprotein F3, which intervenes in axonal outgrowth, and the polysialylated isoform of the neural cell adhesion molecule (PSA-NCAM), which reduces cell adhesion and promotes dynamic cell interactions. F3 is colocalised with vasopressin and oxytocin hormones in neurosecretory granules and follows an activity-dependent, regulated pathway for surface expression on neurohypophysial axons. In contrast, PSA-NCAM appears to follow a constitutive pathway, independent of the activity of the hypothalamo neurohypophysial system, for expression on axonal and glial surfaces, in the hypothalamic magnocellular nuclei and in the neurohypophysis. The role of F3 remains to be determined but in view of its presumptive functions during development, we propose that it promotes remodelling of neurosecretory terminals. On the other hand, we provide direct evidence that surface expression of PSA on NCAM is essential to morphological plasticity since its specific enzymatic degradation in vivo inhibited the neuronal-glial and synaptic changes normally induced by stimulation of secretion from the hypothalamo-neurohypophysial system. PMID- 10795923 TI - Mechanisms of glial retraction in the hypothalamo-neurohypophysial system of the rat. AB - It has been known for more than twenty years that changes in glial coverage of magnocellular neurones in the hypothalamo-neurohypophysial system accompany activation of those neurones. This led to the so-called 'glial retraction hypothesis.' However, until recently, little has been established as to how this structural plasticity of astrocytes develops. This paper will explore a number of hypotheses and supporting data concerning these changes. PMID- 10795924 TI - Genetic models of vasopressin deficiency. AB - Animal models of genetic hormone deficiency are useful as models for physiological studies of hormone deficiency and hormone action. Structure function studies of the specific underlying gene defect may help in understanding mechanisms regulating gene expression and secretion of the peptide product. Spontaneous genetic models of vasopressin deficiency, such as the Brattleboro rat and human familial diabetes insipidus, have facilitated many studies of vasopressin. However, the Brattleboro rat may not be an ideal model of genetic vasopressin deficiency and therefore could be less useful for studies of the central nervous system or as a background strain for the introduction of new vasopressin gene constructs. The human model is appropriately limited by the constraints of human studies, so that engineered animal models of specific diseases, such as familial neurohypophysial diabetes insipidus, are required. The recent development of a vasopressin-null mouse may provide insights into the various roles of vasopressin in the stress response, cardiovascular regulation and behaviour. Additionally, animals with a complete genetic deficiency of vasopressin can serve as a background strain for introduction of novel vasopressin gene constructs to enable sophisticated studies of the regulation of vasopressin expression and the intracellular processes required for appropriate secretion of vasopressin peptide. As advanced techniques of genetic manipulation become more reliable, conditional expression of vasopressin, regulated by time or body site will permit even more detailed studies in this field. PMID- 10795925 TI - Transgenic studies in rats and mice on the osmotic regulation of vasopressin gene expression. AB - Over the past 10-15 years, profoundly important transgenic techniques have been developed that enable new genes to be introduced into whole mammalian organisms. This review describes the ways in which transgenic animals, both rats and mice, have been used to study the mechanisms by which the expression of the vasopressin gene is confined to specific neurones in the hypothalamus, and how the pattern of that expression is altered following an osmotic challenge to the organism. PMID- 10795926 TI - Control of the renal medullary circulation by vasopressin V1 and V2 receptors in the rat. AB - Utilization of the acute and chronically instrumented Sprague-Dawley rat model has provided new and informative data about the mechanisms of, and the role that circulating arginine vasopressin plays in, the regulation of blood flow to the renal medulla. Regional changes of blood flow were measured using implanted optical fibres and laser-Doppler flowmetry techniques. Transcriptional and translational sites of the V1a and V2 receptors were determined in microdissected intrarenal vascular segments from the cortex and medulla. Results from acute and chronic studies indicate the following. First, physiological elevations of plasma vasopressin concentration seen with 48 h of water restriction reduce blood flow to the inner medulla (via V1 receptors) while maintaining a constancy of blood flow to the outer medulla. Reduction of medullary blood flow is necessary to optimize urine osmolality during water restriction. Second, increases of plasma vasopressin concentration of as little as 8 pg ml(-1), which produce no change in baseline arterial pressure or renal cortical blood flow, can lower medullary blood flow selectively and greatly attenuate the arterial pressure-blood flow and pressure-natriuresis relationship. Third, medullary blood flow does not remain reduced in the face of sustained elevations of plasma vasopressin concentration, which appears to be related to the inability of vasopressin to produce a sustained hypertension. Fourth, V1a receptor mRNA and protein are present in the isolated cortical and medullary vasculature, but the V2 receptor mRNA and protein are found only in tubular segments. Levels of V2 receptor mRNA during water restriction were quantified using a competitive RT-PCR and a deletion mutant RNA transcript to control for the efficiency of the reaction, and Western blot analysis was utilized for quantification of the V2 receptor protein. The results demonstrated a time-dependent downregulation of the V2 receptor mRNA and protein within the rat kidney, specifically in the outer medulla. Fifth, the vasopressin induced vasoconstriction of the medullary vasa recta microvessels was shown to be mediated via V1a receptors, and this response is normally modulated by vasopressin-stimulated release of nitric oxide (NO), via extravascular (presumably medullary collecting duct ) stimulation of V2 receptors. Finally, chronic vasopressin administration (10 days) increased nitric oxide synthase activity in the outer medulla and interstitial NO concentration in the medulla. These changes are essential to provide a constancy of blood flow to the renal medulla and buffer against the hypertensive actions of this potent vasoconstrictor peptide. PMID- 10795927 TI - Role of water channels in fluid transport studied by phenotype analysis of aquaporin knockout mice. AB - Aquaporin-type water channels are expressed widely in mammalian tissues, particularly in the kidney, lung, eye and gastrointestinal tract. To define the role of aquaporins in organ physiology, we have generated and analysed transgenic mice lacking aquaporins (AQP) 1, 3, 4 and 5. Multiple phenotype abnormalities were found in the null mice. For example, in kidney, deletion of AQP1 or AQP3 produced marked polyuria whereas AQP4 deletion produced only a mild concentrating defect. Deletion of AQP5, the apical membrane water channel in the salivary gland, caused defective saliva production. Deletion of AQP1 or AQP5, water channels in lung endothelia and epithelia, resulted in a 90% decrease in airspace capillary water permeability. In the brain, deletion of AQP4 conferred marked protection from brain swelling induced by acute water intoxication and ischaemic stroke. The general paradigm that has emerged from these phenotype studies is that aquaporins facilitate rapid near-isosmolar transepithelial fluid absorption/secretion, as well as rapid vectorial water movement driven by osmotic gradients. However, we have found many examples in which the tissue-specific expression of an aquaporin is not associated with any apparent phenotypic abnormality. The physiological data on aquaporin null mice suggest the utility of aquaporin blockers and aquaporin gene replacement in selected human diseases. PMID- 10795928 TI - Renal urea transporters. Direct and indirect regulation by vasopressin. AB - Urea is the most abundant urinary solute and is excreted in urine at a much higher concentration than in other body fluids. Urea concentration is achieved in the kidney through complex urea movements between blood vessels and renal tubules, which involve facilitated urea transport. Three major urea transporters expressed in the kidney have been cloned, UT-A1, UT-A2 and UT-B1, the first two derived from the same gene by differential transcription. These membrane proteins enable facilitated diffusion of urea through specific parts of the nephron (UT-A) and through renal vasculature (UT-B) in the medulla. UT-A1 is localised in the terminal part of the inner medullary collecting ducts and accounts for the vasopressin-dependent increase in urea permeability of this segment. UT-A2 is found in the descending thin limbs of Henle's loops. UT-B1 is expressed in the endothelium of the descending vasa recta supplying blood to the renal medulla, and in red cells. All three urea transporters are primarily involved in the process of intrarenal urea recycling, which enables the establishment, and prevents the dissipation, of a high concentration of urea in the inner medulla. This is an essential feature for producing a concentrated urine and thus for water economy in mammals. Vasopressin, upon binding to V2 receptors in the inner medullary collecting ducts, increases urea permeability through activation of UT A1 molecules, thus enabling urea to diffuse into the inner medullary interstitium. Urea then taken up in ascending vasa recta is returned to the inner medulla via UT-A2 and UT-B1 by countercurrent exchange. These latter two urea transporters are not influenced acutely by vasopressin, but UT-A2 expression is markedly increased in the descending thin limbs of the loops of Henle after sustained exposure to vasopressin or its V2 agonist dDAVP. This effect is indirect because vasopressin receptors are lacking in the descending limbs. The acute direct and delayed indirect actions of vasopressin on renal urea transporters will increase medullary urea accumulation and thus the ability of the kidney to conserve water. Atrial natriuretic peptide inhibits the vasopressin dependent increase in urea permeability in the inner medullary collecting ducts. The interruption of urea recycling probably contributes to the natriuresis. Impairing in this way the capacity of the kidney to concentrate urea enhances its capacity to concentrate sodium in the urine. PMID- 10795929 TI - The treatment of hyponatraemia using vasopressin antagonists. AB - Hyponatraemia is a frequent electrolyte disorder. It is primarily attributable to vasopressin excess plus sustained fluid intake. Hyponatraemia causes CNS symptoms, especially during the first 2-4 days; these symptoms are related to brain swelling. Hyponatraemia occurs in the setting of liver cirrhosis and congestive cardiac failure, in which it is related to stimulation by low arterial blood pressure acting through baroreceptors. Hyponatraemia also occurs in the syndrome of inappropriate antidiuretic hormone secretion, usually from neoplasms releasing vasopressin. The conventional treatment of hyponatraemia used to be fluid restriction and treatment of the underlying disorder. This kind of treatment has been unreliable, cumbersome and difficult to comply with for the patient. In the future, effective vasopressin V2 antagonists will become available for clinical use in the treatment of hyponatraemia, and are expected to improve the management of hyponatraemia. Pharmacological characteristics and observations of biological effects of three antagonists are reported in the present article. PMID- 10795930 TI - Vasopressin receptor antagonism--a therapeutic option in heart failure and hypertension. AB - The precise role of vasopressin in the pathophysiology of cardiovascular disease is controversial, but this peptide hormone is important for several reasons. Firstly, circulating concentrations of vasopressin are elevated in heart failure and some forms of hypertension. Secondly, there is evidence that vasopressin is synthesized not only in the hypophysial-pituitary axis but also in peripheral tissues including the heart where it acts as a paracrine hormone. Thirdly, vasopressin has vasoconstrictor, mitogenic, hyperplastic and renal fluid retaining properties which, by analogy with angiotensin II, may have deleterious effects when present in chronic excess. Finally, the availability of orally active non-peptide vasopressin receptor antagonists allows vasopressin receptor antagonism to be considered as a therapeutic option in cardiovascular disease. PMID- 10795931 TI - Quo vadis neurohypophysial hormone research? AB - Here we highlight just a few of the outstanding questions in the field of neurohypophysial hormones that we envisage will be addressed successfully in the new millennium. To begin, we focus on the regulation of receptors. Despite intensive investigation with new drugs, molecular modelling and transgenic models, the determinants of receptor selectivity remain elusive; there may even be more vasopressin or oxytocin receptor subtypes to be discovered. We discuss the controversy over the interesting studies that indicate modulation of oxytocin receptor-binding by steroids. Oxytocin and vasopressin release and action in the brain are discussed from several aspects. Dendritically released oxytocin acting locally is important for the milk ejection reflex, and similarly released vasopressin is important in regulating patterning of vasopressin neurone activity. Such dendritically released oxytocin and vasopressin is likely to be important in paracrine modulation of neural circuitry involved in neuroendocrine control, and for a range of behaviours. Is it possible that the whole range of behaviours that comprise 'social' (or 'anti-social') or 'maternal' behaviour can be engineered by modifying the expression of just these one or two peptides and their receptors? However, whether gene expression and knockout approaches will answer all the open questions about the real functions of oxytocin and vasopressin remains to be shown. PMID- 10795932 TI - Intra-operative spinal cord monitoring in orthopaedics. AB - The surgical correction of spinal deformities carries a small but significant risk of injury to the spinal cord. To detect the onset and possibly reverse the effects of surgical complication, a variety of neurophysiological monitoring procedures can be employed. The purpose of this review is to provide information regarding the various methodologies available for monitoring spinal cord and nerve root function during orthopaedic procedures. Intra-operative monitoring of cortically recorded somatosensory evoked potentials (SEPs) by peripheral nerve stimulation is of value during orthopaedic surgery and is the state-of-the-art in terms of non-invasiveness, versatility, time requirement, lateral discrimination, and ease of electrode placement. Monitoring of motor evoked potentials (MEPs) is useful particularly in combination with SEPs but is still considered investigational. Root function monitoring has limited application and requires more clinical research. PMID- 10795933 TI - Surgical management of incomplete cervical cord injury with stenosis secondary to ossification of the posterior longitudinal ligament. PMID- 10795934 TI - Total spondylectomy for primary tumor of the thoracolumbar spine. AB - STUDY DESIGN: Six patients with primary malignant tumor of the thoracolumbar spine who underwent total spondylectomy (TS) by en bloc resection were reviewed retrospectively. OBJECTIVES: To report surgical technique and preliminary results of TS and to evaluate its oncological curability. SETTING: Japan. METHODS: Six patients were treated by TS by en bloc resection of the vertebral tumor. TS through a posterior approach was performed in three cases (T1 osteosarcoma, L1 osteosarcoma and L1 chordoma) and in the others through a single stage anterior and posterior combined approach (T6-8 recurrent giant cell tumor. L4 chordoma and L5 giant cell tumor). Surgical margins of the specimens were evaluated histologically. All patients were followed, and their status was evaluated by clinical and imaging studies. RESULTS: There were no complications related to surgery. Programmed sacrifice of nerve roots were performed in three cases for oncologic excision. A wide surgical margin was achieved in one case, a marginal one in four, and an intralesional margin in one. Five patients were alive without evidence of tumor and one was alive with disease at follow-up evaluation after 2.0-4.8 years. Local recurrence was found in one case of T1 osteosarcoma with an intralesional margin. CONCLUSIONS: These preliminary results suggested that TS is an effective procedure in control of local recurrence with acceptable complications. PMID- 10795935 TI - Microdialysis of the spinal cord during thoracic aortic cross-clamping in a porcine model. AB - OBJECTIVE: Utilising microdialysis to measure the changes of glucose, lactate, pyruvate and glutamate levels in the spinal cord after cross-clamping of the thoracic aorta in an established porcine model to monitor the degree of ischaemia. DESIGN: Experimental study with a porcine model. SETTING: University Hospital, Trondheim. SUBJECTS: Six pigs. MAIN OUTCOME MEASURE: Lactate, pyruvate and glutamate concentrations in the microdialysis perfusate from the spinal cord. RESULTS: A significant increase of the lactate-pyruvate ratio during the last 30 min of the 1 h clamping period, with a maximum increase of 169% from the basal value the last 10 min before declamping. No evident change in this ratio between the clamping and the reperfusion period. No significant change in glutamate levels during clamping or reperfusion period. CONCLUSION: Microdialysis reflects the ischaemic state of the spinal cord during cross-clamping of the thoracic aorta in pigs, and is well suited to study each phenomena. PMID- 10795936 TI - Systemic circulation and cerebral oxygenation during head-up tilt in spinal cord injured individuals. AB - OBJECTIVE: To compare tilt-induced alterations in cardiovascular homeostasis and cerebral oxygenation of spinal cord-injured (SCI) to able-bodied (AB) individuals. DESIGN: Subjects underwent 10 min supine rest followed by 10 min 70 degrees head-up tilt. The last 5 min of supine rest and head-up tilt were analyzed, provided a steady state existed. SUBJECTS: SCI individuals (n= 11), with lesions between C4 and T4, and AB individuals (n= 10), all males and balanced for age and weight. MAIN OUTCOME MEASURES: Calf circumference, mean arterial pressure (MAP), stroke volume, heart rate and cerebral oxygenated ([O2Hb]), deoxygenated ([HHb]) and total ([tHb]) haemoglobin concentration changes were measured. RESULTS: Head-up tilt evoked a greater fall in MAP (mean (SD): -9 (12) vs 2 (6) mmHg P=0.02) and stroke volume (-43 (12) vs -22 (10)%, P=0.005), and a greater increase in heart rate (27 (12) vs 18 (6) beats, P=0.04) in SCI than AB. Cardiac output decreased during head-up tilt in SCI but not in AB (-17 (15) vs 1 (15)%, P=0.01). The change in cerebral oxygenation ([HHb]: 3.9 (2.8) vs 2.8 (1.4) micromol x l(-1), P=0.1 and [O2Hb]: -6.1 (5.0) vs -2.1 (5.5) micromol x l(-1), P=0.1) was similar in SCI and AB. All variables mentioned showed a change significantly different from zero in both groups, apart from [O2Hb] in AB and [tHb] in both groups. CONCLUSION: SCI demonstrated a greater decrease of MAP and stroke volume with a similar decrease in cerebral oxygenation compared to AB. This suggests that although systemic circulation was less well regulated in SCI compared with AB, cerebral circulation in SCI was maintained as in AB. PMID- 10795937 TI - The Tetrafigap Survey on the long-term outcome of tetraplegic spinal cord injured persons, part II: Demographic characteristics and initial cause of injury. AB - OBJECTIVES: After presenting in part I the protocol and the methodology of the Tetrafigap Survey, this article is aimed at investigating demographic characteristics, initial cause of trauma and spinal injury in the French tetraplegic spinal cord injured (TSCI) population. STUDY DESIGN: The Tetrafigap Survey is a multicentre epidemiological survey on the outcome of TSCI people following their first admission to a rehabilitation department or centre. SETTING: The survey was conducted with the participation of 35 French-speaking collaborating centres. METHOD: A self-administered questionnaire was carried out among consenting TSCI patients followed by the centres. 1,668 questionnaires fulfilled all the required criteria for data analysis. RESULTS: The socio demographic variables (gender, current age and age at the time of accident, duration of disability), circumstances of accident as well as initial spinal cord lesions are reviewed in the body of the text. CONCLUSION: The results from this survey form the first detailed description of the French TSCI population. Whereas demographic data show many similarities with those of previous studies in other countries, some specific differences are highlighted, in particular relating to the type of accident. PMID- 10795938 TI - Sports, recreation and employment following spinal cord injury--a pilot study. AB - STUDY DESIGN: A pilot postal survey. OBJECTIVE: To examine the levels of sporting/recreational activities, education and employment in people with spinal cord injury (SCI) and to assess if involvement in sport and recreation is associated with higher levels of education and employment. SETTING: National Spinal Injuries Centre (NSIC), Stoke Mandeville Hospital, Aylesbury, UK. METHODS: Forty-five subjects, who met the following criteria: SCI at level C5 or below, wheelchair dependent, aged between 18 and 50 at the time of injury, injured at least 10 years ago, admitted to the NSIC within 6 months of injury and resident in the UK, completed a 26-item postal questionnaire. RESULTS: Among the 45 subjects 47% participated in physical activities (20% in sport; 27% in recreation), 33% were employed (29% full-time) and 18.5% had undergone further education at the time of the survey. Participation in sports as well as employment status decreased after injury (P<0.01) usually as a result of poor access to sports and work facilities. Only 4% of investigated SCI patients started to practise physical activity after the accident whilst 42% stopped. CONCLUSIONS: Levels of sporting/recreational activities and employment decreased significantly after injury. No significant correlation was found between sport/recreation involvement and education/employment status. Further investigation with a large number of participants that will enable additional analysis of subgroups, such as level of injury and functional independence, is required. PMID- 10795939 TI - Prediction of functional outcome after spinal cord injury: a task for the rehabilitation team and the patient. AB - STUDY DESIGN: Descriptive analysis of data gathered in an information system. OBJECTIVES: To explore the predictions of professionals and patients regarding functional outcome after spinal cord injury related to the final results after inpatient rehabilitation, in order to make prognostics of rehabilitation outcome more successful and enlarge the role of the patient in selecting realistic rehabilitation goals. METHODS: Data from 55 patients with spinal cord injury admitted to the rehabilitation centre. Expectations of the rehabilitation team and the patients regarding future independence in performing six daily activities were compared to the functional results at discharge. The results of patients with different level and extent of lesion were analyzed. RESULTS: In 52% of all performed skills, independence was achieved at discharge. Professionals and patients made similar predictions. If they both expected independence after rehabilitation. 90% of the skills were performed independently at discharge. If they both did not expect independence only 3% of the functional results were positive. Of all combined predictions 64% was correct. Correct predictions were most often found regarding self-care skills of patients with paraplegia and regarding mobility of patients with complete lesions. Prediction of self-care outcome of patients with tetraplegia is far more complicated. There was a considerable variation in predictions of mobility potential, especially regarding patients with incomplete lesions. If the team and patients agreed upon expected independence in mobility skills of these patients, the final results were mostly positive. CONCLUSIONS: Prediction of functional outcome after spinal cord injury was most successful if the expectations of the team and patients were combined. Prognosis of self-care outcome of patients with paraplegia and mobility potential of patients with complete spinal cord lesions was usually clear at admission. However, selection of realistic goals concerning self-care skills of patients with tetraplegia and mobility skills of patients with incomplete lesions is far more complicated. Gradual adjustment of objectives is needed during the rehabilitation process in close collaboration between the professionals and the patients. PMID- 10795940 TI - Spontaneous spinal subdural hematoma with spontaneous resolution. AB - STUDY DESIGN: Single patient case report. OBJECTIVES: To present and discuss a case of spontaneous spinal subdural hematoma that was not associated with coagulation abnormality. SETTING: Seoul National University Hospital, Seoul, Korea. METHODS: Clinical and radiological magnetic resonance imaging follow-up of the patient between August 1997 and December 1998. RESULTS: The hematoma was initially extensive, ranging from T5 to L3. However, it resolved spontaneously with the subsequent development of an adhesive arachnoid cyst. CONCLUSION: Even though urgent surgical decompression is necessary in most cases of spinal subdural hematoma, conservative management could be an option in cases with rapid resolution of the neurological deficit. PMID- 10795942 TI - Post-traumatic syringomyelia. PMID- 10795941 TI - Dosage of once-daily gentamicin in spinal cord injury patients. AB - INTRODUCTION: In patients with spinal cord injury (SCI), serum creatinine does not accurately reflect the level of renal function. Therefore, in SCI patients, the dose of potentially nephrotoxic drugs should be adjusted on an individual basis from the estimated creatinine clearance. CASE REPORT: A 41-year-old male with tetraplegia due to cervical spinal cord injury underwent extended pyelolithotomy for staghorn calculus in the right kidney. The blood urea level was 9.9 mmol/l; creatinine was 112 umol/l (reference range: 0-135). We were conscious of this patient's renal disease, and therefore, administered only 3 mg/kg of gentamicin (240 mg) instead of the standard dose of 5 mg/kg body weight. Despite taking this precaution, the gentamicin level measured 22.5 h after the initial dose, was in the potentially toxic range 3.3 mg/l. CONCLUSION: We recommend that even the first dose of gentamicin in the once-daily regimen, which is 5 mg/kg, should be individualised in SCI patients based on age, sex, weight, height, level of spinal cord injury, and renal function. PMID- 10795943 TI - Multidimensional subtypes and treatment outcome in first-time DWI offenders. AB - The concurrent and predictive validity of Type A and B alcoholism subtypes was evaluated in 246 first-time driving-while-intoxicated (DWI) offenders. K-means analysis indicated that a two-cluster solution was optimal with Type Bs (28%) exhibiting greater premorbid risk factors, alcohol and psychosocial severity, drinking consequences, psychopathology, higher stage of change, and less coping confidence in comparison to less severe Type As (72%). After baseline assessment, participants were randomly assigned to one of three 10-week group treatments (DWI Education, Coping Skills, Interactional), and reassessed at termination, and at 6 month and 1-year follow-ups. Type B was associated with more severe symptoms after treatment, but there was no evidence for patient-treatment matching effects. Although Type A/B may be an important theoretical model for guiding alcoholism research, it usefulness and efficiency for treatment matching, planning, or placement purposes is questioned. PMID- 10795944 TI - Cluster subtypes within stage of change in a representative sample of smokers. AB - In this paper a three-level conceptualization of approaches to matching self-help behavior change strategies to individuals is presented based on the Transtheoretical Model of Change. Two of the model's intervention approaches, tailoring to Stage-of-Change and creating completely individualized interventions, have already been developed and implemented in previous research. A third intervention approach, in between stage-tailored and individualized, is targeting subtype groups within each Stage of Change. The subtype targeted intervention approach is in the initial development stage of empirically determining and validating the stage subtypes. Three studies are presented which investigate stage subtypes within a representative sample of 4,144 smokers in the Precontemplation, Contemplation, and Preparation Stages of Change. Within each Stage-of-Change study, two cluster analyses were performed using the Pros, Cons, and Situational Temptations from the Transtheoretical Model to establish cluster replicability. Cluster solutions were externally validated using the 10 Processes of Change and 2 smoking behavior variables. Four distinct subtypes were found in Precontemplation and Contemplation, and five subtypes were found in Preparation. These subtypes closely replicate subtypes previously found in a convenience sample of smokers and provide strong evidence for the existence of subtypes within the first three Stages of Change. PMID- 10795945 TI - Differences in daily stress, mood, coping, and eating behavior in binge eating and nonbinge eating college women. AB - This study investigated differences between binge eaters and controls on measures of daily stress, mood, coping, and eating behavior. The same measures were also used to compare binge days to nonbinge days for the binge group. Participants included 20 college women who reported binge eating at least two times per week and 20 women who reported no disturbance in their eating behavior. They were asked to self-monitor their daily stress, coping, mood, and eating behavior for three weeks. Results indicated that the binge group reported more stress and negative mood over the three weeks than the control group. The binge group reported experiencing a similar number of stressful events on binge days as compared to nonbinge days, however, the impact of those events was much greater on binge days. The binge eaters also reported less positive mood and more episodes of eating on binge days. The groups did not differ in the number of coping strategies used. Stress and negative mood appear to be common antecedents for binge eating. The role of coping responses and daily eating behavior (i.e., restraint) on binge eating is somewhat less clear. PMID- 10795946 TI - Ethical issues in research on the treatment of adolescent substance abuse disorders. AB - Treatment research on adolescent substance use disorders raises a number of important ethical and legal concerns which have not been widely acknowledged. This paper explores these concerns as they relate to fundamental ethical principles in the conduct of human research. The issues discussed include tensions between conflicting regulations governing informed consent for research and treatment of adolescents, the capacity of adolescents to give informed consent, potentially coercive elements related to research on substance use treatment, problems associated with confidentiality and release of information, research vulnerability associated with substance use, and ethical implications of distinctions between effectiveness and efficacy research. Suggestions for ways investigators may address these concerns are provided. PMID- 10795947 TI - Self-efficacy and motivation for controlling drinking and drinking/driving: an investigation of changes across a driving under the influence (DUI) intervention program and of recidivism prediction. AB - Measures of (a) self-efficacy and (b) motivation to change (stage) for controlling drinking and drinking/driving were examined at the beginning and the end of a four-week intervention in a sample of 670 Driving Under the Influence (DUI) offenders in a court-mandated program. Hypotheses regarding stability of stage classifications over the course of intervention, and the relation between stage classification, stage scores, self-efficacy, and DUI recidivism were examined. Based on results of an earlier study it was expected that most offenders would be classified into the action stage at entry and that classifications would tend to remain stable from pretest to posttest. Action was the most frequent stage classification in both drinking and drinking/driving domains at both test periods, with precontemplation being the least frequent classification. When tracked over the four weeks, stage classifications for drinking and drinking/driving were stable for 74 to 89% of offenders in the two domains, respectively. As predicted, higher action and self-efficacy scores were related to lower recidivism, and action scores in the drinking/driving domain were the best early recidivism predictors among a predictor set that included traditional recidivism indicators. Drinking contemplators (i.e., those with the highest stage score on the contemplation scale) had higher recidivism rates than other drinking stage classifications. Implications for DUI intervention programs are discussed. PMID- 10795948 TI - The transtheoretical model of smoking: comparison of pregnant and nonpregnant smokers. AB - Pregnant smokers represent a special population of women smokers. Since smoking affects both the mother and the unborn child, it might be hypothesized that pregnant women should be highly motivated to quit smoking during pregnancy. Results from both naturalistic studies and intervention studies have generally shown only modest changes in smoking during pregnancy. To help design more effective interventions tailored for this special population of smokers, more information is needed on how they differ from other smokers. The current study was designed to examine patterns and differences on constructs of the Transtheoretical Model between low-income culturally diverse pregnant and nonpregnant female smokers. Groups were compared on the distribution of the stages of change. In addition, stage and group comparisons were conducted for the pros and cons of smoking, situational temptations to smoke, and the processes of change. The sample included 103 economically disadvantaged, culturally diverse pregnant women and a matched group of 103 nonpregnant women smokers. Pregnant smokers as a group were similar to their nonpregnant peers on their readiness for quitting. In general, the patterns observed across the stages of change were consistent with those seen in other populations of smokers. The primary difference found between the groups were on the cons of smoking, habit-related temptations to smoke, and experiential processes of change. Specifically, pregnant women perceived a less negative attitude toward their smoking, were more tempted in habit-related situations, and made less use of important experiential processes of change. The implications of these findings for designing interventions tailored for this special group of smokers are discussed. PMID- 10795949 TI - Co-occurrence of alcohol, smokeless tobacco, cigarette, and illicit drug use by lower ranking military personnel. AB - The Worldwide Survey of Health Related Behaviors is administered periodically to a probability sample of military personnel. Earlier reports of these surveys suggested that illicit drug use was highest among the lowest ranking personnel. This paper reports a secondary analysis of the 1992 and 1995 surveys of the lowest ranking personnel. The results suggested that in general illicit drug users tended also to use alcohol, smokeless tobacco, and cigarettes. Heavy drinkers were more likely than light drinkers to use illicit drugs. No such relationship was observed between illicit drug use and the level of use of cigarettes or smokeless tobacco. Moreover, among the heavy drinkers, illicit drug users were especially likely to use cigarettes and among males, smokeless tobacco. The relevance of these results to military policies toward illicit drug use is discussed. PMID- 10795950 TI - Improving treatment outcomes for pregnant drug-dependent women using low magnitude voucher incentives. AB - The aim of this study was to examine the effectiveness of low-magnitude behavioral incentives in improving attendance for abstinence-treated patients and sustaining illicit-drug abstinence for methadone-treated patients. Subjects were randomly assigned to either incentive or control conditions, with target behaviors differing for the two patient groups (attendance for abstinence-treated and abstinence for methadone-treated patients). Controls received no incentives, whereas incentive subjects could earn $5/day in vouchers during the first 7 days of an intensive outpatient treatment. Results showed that $5/day did not significantly improve attendance in abstinence-treated patients or impact drug abstinence in methadone-treated patients. The data suggest that low-magnitude voucher incentives enhanced treatment attendance by methadone-treated subjects. Although modest monetary incentives had some utility in improving attendance in methadone-treated patients, more potent interventions are needed to improve attendance and maintain abstinence in this high-risk population. PMID- 10795951 TI - Relationships between drinking motives and drinking restraint. AB - Relationships between drinking motives (self-perceived reasons for drinking alcohol) and drinking restraint (preoccupation with controlling alcohol intake) were examined in a nonclinical young adult sample. Ninety-seven undergraduate university drinkers completed the Temptation and Restraint Inventory (Collins & Lapp, 1992), the Drinking Motives Questionnaire (Cooper, Russell, Skinner, & Windle, 1992), and measures of demographics (age and gender) and social desirability. Results indicated that after accounting for the influences of demographic and social desirability information, Coping Motives and Enhancement Motives scores from the Drinking Motives Questionnaire were significant predictors of Cognitive and Emotional Preoccupation scores on the Temptation and Restraint Inventory, and Coping Motives scores were a significant predictor of Cognitive and Behavioral Control scores on the Temptation and Restraint Inventory. Social Motives scores on the Drinking Motives Questionnaire did not significantly predict either Cognitive and Emotional Preoccupation or Cognitive and Behavioral Control scores. Further analyses suggested that actual behavioral attempts at alcohol restriction on the Temptation and Restraint Inventory were predicted by Enhancement Motives scores, whereas cognitive concerns about drinking were predicted by Coping Motives scores. Results are discussed in terms of implications for risk for excessive and problem drinking in enhancement and coping-motivated young adult drinkers. PMID- 10795952 TI - Cigarette smoking among immigrant Salvadoreans in Washington, DC: behaviors, attitudes, and beliefs. AB - There has been an influx of immigrants from El Salvador to the Washington, DC (DC) area, but little is known about the health behaviors of this population. This study was conducted to describe the prevalence of cigarette smoking among adult Salvadorean immigrants to the DC area. Bilingual interviewers administered a face-to-face interview to participants recruited from throughout the community. Complete data were available for 1,458 participants: 10.8% of those surveyed were current smokers and 11.7% were former smokers. Men were significantly more likely than women to have ever smoked either in the past (adjusted prevalence difference [PD = 21.0%] or currently (PD = 21.2%). The respondents tended to believe that smoking was a "habit" rather than an addition. Only 16% lived in households where smoking was permitted, and the majority supported smoke-free policies in public places, with men and current smokers being less permissive. The smoking behavior exclusively represented the smoking pattern that the Salvadoreans had adopted before immigration. The data suggest that smoking control strategies aimed at this population should seek to reduce the onset of smoking among men and continue to keep smoking among women rare. PMID- 10795953 TI - The effect of self-discrepancy and discrepancy salience on alcohol consumption. AB - The effect of self-discrepancy magnitude and salience on alcohol consumption was examined in an ad lib drinking study in order to evaluate the utility of the self inflation component of the myopia model for better understanding drinking practices. Participants were 33 males and 27 females recruited on a university campus. It was predicted that participants with relatively large real self/ideal self discrepancies on dimensions important to their self-concept would consume the greatest amount of alcohol in a wine tasting test. Moreover, this effect was expected to be enhanced when self-discrepancies were made salient. The results of hierarchical regression analyses showed a main effect of gender and a significant interaction between self-discrepancy magnitude and salience condition. However, the interaction was such that wine consumption tended to decrease as discrepancy magnitude increased in the condition in which self-discrepancies were made salient, with the opposite relationship in the control condition. Three possible reasons for the unexpected findings are discussed: (a) The salience manipulation did not perform as expected: (b) the sample had little to gain from self inflation: and (c) typically, self-inflation does not significantly motivate alcohol consumption. PMID- 10795954 TI - Minors' access to snuff: purchase outcomes at corporate owned and independently owned convenience stores. AB - Results from many studies indicated that sales clerks sell cigarettes to minors. However, not many researchers have investigated how frequently merchants sell smokeless tobacco to minors. In the present study, sales of snuff to minors were assessed at corporate owned chains of convenience stores and at independently owned convenience stores. Two corporate owned chains with the largest number of convenience stores were selected (n = 45). A random sample of 45 independently owned convenience stores that held current tobacco licenses were also chosen. Two underage males attempted to purchase moist snuff at each of the 90 convenience stores included in the sample. Overall, underage buyers were sold snuff on one out of four purchase attempts. Sales occurred significantly more often in independently owned stores (35.6%) than in corporate owned stores (13.3%). An important variable that appeared to determine whether the sales clerk sold snuff was request for identification. PMID- 10795955 TI - Alcohol expectancies and social deficits relating to problem drinking among college students. AB - Standardized questionnaires were administered to 116 male and female undergraduates to examine how social deficits and alcohol expectancies relate to alcohol use. Participants were classified as either problem or nonproblem drinkers based on the Rutgers Collegiate Substance Abuse Screening Test. Problem drinkers reported experiencing social anxiety, shyness, and lower self-esteem to a greater extent than nonproblem drinkers. Problem drinkers also held more positive alcohol expectancies than nonproblem drinkers. Contrary to our hypotheses, however, particular types of alcohol expectancies did not interact with specific areas of social functioning to influence problem drinking. Overall, these findings suggest that problem drinkers have positive expectations about the immediate effects of alcohol consumption even though drinking is linked to long term impairment in social functioning. PMID- 10795956 TI - Effect of three smoking cessation treatments on nicotine withdrawal in 141 abstinent alcoholic smokers. AB - This prospective study examined the effect of three behavioral smoking interventions and reductions in cigarettes smoked per day on nicotine withdrawal symptoms in 141 abstinent alcoholic smokers (73 men, 68 women). The participants' mean +/- SD age was 41.4 +/- 9.2 years. They smoked an average of 27.7 +/- 12.1 cigarettes per day and reported 4.1 +/- 4.3 years of current abstinent from alcohol and other drugs of dependence. Participants were randomly assigned to a 12-week program of standard treatment (ST, n = 61), behavioral counseling plus exercise (BEX, n = 39), or behavioral counseling plus nicotine gum (BNIC, n = 41). All three conditions included instructions to reduce the number of cigarettes smoked per day prior to the target quit date (TQD). The TQD was week 4 for ST subjects and week 8 for those in the BEX and BNIC groups. The post treatment assessment occurred one week after TQD. The Profile of Mood States (POMS) and the Beck Depression Inventory were administered at baseline and posttreatment to assess nicotine withdrawal. Significant increases were detected for the POMS total mood disturbance score, and the depression, tension, anger and confusion subscales, while vigor scores decreased (all p < 0.03). Withdrawal change scores were not found to be associated with treatment condition or percentage reduction in cigarettes, and there was no evidence of a significant interaction of treatment and cigarette reduction. Results are discussed in relation to implications for treatment and for future research. PMID- 10795957 TI - Fluoxetine versus placebo in depressed alcoholics: a 1-year follow-up study. AB - The authors conducted a first study to evaluate the long-term efficacy of fluoxetine for decreasing the depressive symptoms and the drinking of patients with comorbid major depressive disorder and alcohol dependence. This study consisted of a 1-year naturalistic follow-up of 31 patients who previously had completed a 3-month double-blind, placebo-controlled study of fluoxetine in depressed alcoholics. The fluoxetine group continued to demonstrate less depressive symptoms and less drinking than the placebo group at the 1-year follow up evaluation. The results of the 1-year follow-up evaluation suggest persistent efficacy for fluoxetine for treating the depressive symptoms and the drinking of depressed alcoholics. PMID- 10795958 TI - Depression and smoking cessation: the role of motivation and self-efficacy. AB - The aim of this paper is to examine whether depression, assessed using the Beck Depression Inventory, is related to smoking cessation motivation and self efficacy. In a population sample of 1,547 Finnish men and 1,856 Finnish women, aged between 25-64 years, there were 34% male and 21% female current smokers. The current smokers had higher mean depression scores than former or never smokers. The higher depression scores were related to greater motivation to quit smoking among female smokers [OR = 1.51 (1.15-1.97)]. In addition, higher depression scores were related to lower smoking cessation self-efficacy, especially among male smokers [OR = .63 (.45-.90)]. Depression scores were not related to whether a smoker had had previous cessation attempts. Our results suggest that depression is related to lower smoking cessation self-efficacy while smokers with higher depression scores seem to be more motivated to quit smoking. PMID- 10795959 TI - Factors associated with untreated remissions from alcohol abuse or dependence. AB - This paper describes an epidemiologic-based sample of individuals who remitted from alcohol abuse or alcohol dependence, both with and without treatment, to abstinence or moderate drinking. Inspection of the severity, onset, and duration of alcohol problems experienced by these individuals suggests that there may be two primary categories of drinkers with distinct pathways to remission. The first is a population of individuals who experience significant problems for an extended period of time who then resolve to abstinence through the use of treatment services. The second is a population of individuals who drink heavily at some point in their lives, experience some problems, and then "mature out" of this stage in their life as they age and take on other life roles. PMID- 10795960 TI - HIV/AIDS social and behavioural research: past advances and thoughts about the future. AB - This paper is an introduction to the various contributions in this special issue of Social Science & Medicine which are an attempt to synthesise the main debates of the 2nd European Conference on Social and Behavioural Research on AIDS held in Paris, in January 1998. The paper discusses how the recent advent of highly active antiretroviral therapies (HAART) and new trends in the epidemic (its concentration in the socially most vulnerable groups and countries) have affected the research agenda of European social and behavioural sciences (SBS) in HIV/AIDS. Questions which had already been thoroughly studied by SBS (like determinants of HIV-related risk behaviours, or impact of gender and socio economic inequities as well as discrimination on the diffusion of HIV) will have to be "revisited" in light of these recent changes. New issues (such as risk behaviours among already infected patients. impact of therapeutic advances on psychosocial and daily life management of their disease by people living with HIV/AIDS, adherence to treatment, or "normalisation" of AIDS public policies) will have to be strongly and quickly dealt with, in order for SSB to keep the pace with the rapid evolution of the epidemic and of the societal responses to it. Finally, the paper argues that to face these challenges, new theoretical and methodological advances will have to go beyond the classical oppositions in internal debates among SSB between individualistic and holistic approaches, or between radical criticism of the existing state of the world and practical involvement in public health decision-making. PMID- 10795961 TI - Research on HIV sexual risk: social relations-based approach in a cross-cultural perspective. AB - In opposition to individual-oriented approaches to sexual risk taking, new theoretical approaches have emerged that strive to encompass all of the social relations--that is to say, the connections that structure the components of relations between groups and go beyond simple adaptations to a given interaction- that prevail in a sexual relationship. This paper examines the strengths and weaknesses of such an approach for understanding HIV risk-related behaviour. The analysis is based on data from European surveys of sexual behaviour in the general population, with special attention paid to the data from the Belgian survey, which used a relation-based approach to risk-related behaviour. The analysis shows that sexual behaviour and preventive behaviour patterns of men and women appear to be linked to women's status in a given society. The data also tend to show that the social and preventive contexts also help structure the relations that develop between partners when it comes to negotiating about using a condom. Finally, the same people behave differently depending on the relationship's context. In particular, the balance of power within the couple, which is strongly interconnected with gender relations issues, and even characteristics of the former relationship help explain these behaviour differences. PMID- 10795962 TI - Social change and HIV in the former USSR: the making of a new epidemic. AB - This paper describes the development of the recent HIV epidemic in countries of the former Soviet Union. It explores the socio-political and economic roots of an injection-drug-driven HIV epidemic associated with a drug culture that facilitates HIV transmission. This review, based on many unpublished reports, studies and field notes, discusses the new social and health context in which the epidemic is developing. The evidence of a growing number of drug users in the region is discussed and drug injection behaviour described. The authors present selected data on the heterosexual transmission of HIV and linkages between the drug-linked HIV epidemic and its further spread into the non-injecting population. The potential overlap with the still uncontrolled syphilis epidemic that began in 1990 is probably a key factor in the future of HIV spread in the region. Until now, HIV infection among STD patients has been extremely low. However, rapid changes in sexual norms and behaviours, the growing commercial sex industry, and increased mobility soon may affect the current situation. The huge economic and socio-political crises currently affecting the region have created a "risk situation" for the spread of HIV. Concerns are raised about the appropriateness and the scope of government and non-governmental approaches to the exploding HIV and STD epidemics. PMID- 10795963 TI - Revisiting the concept of 'vulnerability'. AB - The terms 'vulnerable' and 'vulnerability' are used more and more frequently in the areas of both social science research into and prevention of HIV/AIDS, but certain difficulties arise when it comes to applying this concept to actual situations at the heart of which individuals or groups are more exposed to HIV. The concept of vulnerability must thus be clarified to reinforce its heuristic capacity and political and practical relevancy. The first part of this paper is devoted to presenting a heuristic matrix of vulnerability, used in previous research among people living with HIV/AIDS (PWHAs) and to extracting three levels of intelligibility, that is to say, first the social trajectory level, then the level on which two or more trajectories intersect, and finally that of the social context. Each of the elements belonging to these three levels must be described both objectively and subjectively. The identity construction processes are then proposed as particular observation and 'gelling' points for these various levels taken as a whole. In the second part of the paper, we have reviewed how the concept of 'vulnerability' has been defined and used in other fields, notably disaster, famine, and mental health, paying special attention to the crucial points in the debates that are raging in these fields. We have also shed light on a few concepts that are frequently associated with vulnerability, such as victimization, insecurity, and risk. In the third part, we have summarized our approach to vulnerability as a relevant concept for elucidating risk-taking processes and designing intervention programmes. The importance of analysing the inter-individual differences, the variability in time and the relational dimension of all social vulnerability has been stressed. PMID- 10795964 TI - HIV-positive people, risk and sexual behaviour. AB - For a long time, the sexual behaviour of HIV-infected persons did not receive any serious attention for a variety of reasons. Initially, diagnosis of HIV-infection appeared to imply a death sentence. In this context, the sex life of those infected seemed a secondary issue making prevention focused on sexual behaviour hard to imagine. Furthermore, the conviction that stigmatisation should be avoided also precluded an interest in the sexual behaviour of HIV-infected persons. From an epidemiological perspective and in the context of the developments in the medical treatment of AIDS it is important to address the sexuality of HIV-infected people. The scarce research done until now shows that there are various ways in which an HIV-infection affects people's sexuality. It seems that the sexuality of HIV-infected people can be compromised by their infection, inducing various sexual problems. Research also shows that there are HIV-infected people who do engage in unprotected sex, just as there are HIV negative people or people with unknown serostatus who do so. Studies into the determinants of unsafe sex in HIV-infected people suggest that to some extent the same determinants are operative as among people in general. These include intention and self-efficacy regarding safe sex. Recreational drug use also affects safe sex regardless of serostatus. However, safe sex as well as sex in general is different for seropositive persons than for people who are seronegative or have an unknown serostatus. Among seropositive people, sex is also related to dilemma's involving disclosing their serostatus to potential sex partners, and their motivation to protect their partners as well as themselves against surinfection and STD. Furthermore, having to cope with a serious disease induces negative mood states (particularly depression) and may compromise sexual functioning. Comprehensive prevention aimed at HIV infected persons should address these various issues and should be an integrated part of general HIV prevention. PMID- 10795965 TI - Everyday life with AIDS/HIV: surveys in the social sciences. AB - Research on persons infected with HIV is critically assessed by using the notions developed in the sociology of illness. Has this research stimulated new questions or led to reformulations in research paradigms? Methodological problems related to data collection are pointed out, namely: the recruitment of HIV-positive participants for such studies and the stance they adopt during interviews. The contributions made by studies that have focused on adjustment strategies or resorted to the notions of stigmatization, identity and illness trajectory are also discussed. In conclusion, questions are raised about what implications the new HIV/AIDS treatments will have on this field of social science research. PMID- 10795966 TI - Adherence to HIV combination therapy. AB - The emergence of drug-resistant strains of HIV virus and treatment failure can result from non-adherence to antiretroviral therapy. While non-adherence to therapy is not a new issue or specific to HIV/AIDS, it has received renewed attention because of the complicated combination treatment regimens being prescribed. This paper reviews the relevant background literature on the contributions of social and behavioural science to non-adherence to HIV medications. Data indicating problems with adherence prior to combination therapy are reported. Despite limitations, even self-report assessments have already succeeded in showing that adherence to combination therapy is significantly related to HIV viral load. Recent research data are discussed. Implications of findings for counselling patients to increase their adherence are presented. PMID- 10795967 TI - The normalization of AIDS in Western European countries. AB - The occurrence of AIDS led in every Western European country to exceptional innovations in prevention, patient care, health policy and questions of civil rights. This exception can be explained above all by the fact that not only was a health catastrophe feared but also civilizational harm in the field of civil rights. Despite national differences, this brought about similar exceptionalist alliances consisting of health professionals, social movements and those affected. With the failure of a catastrophe to arise, signs of fatigue in the exceptionalist alliance and increasing possibilities of medical treatment, exceptionalism in Europe is drawing to a close. Four phases are distinguished between in this process, given nationally different patterns of development: Approx. 1981-1985: emergence of exceptionalism. The reasons underlying exceptionalism are investigated. Approx. 1986-1991: consolidation and performance of exceptionalism. The investigation centers on the exceptionalist policy model. Approx. 1991-1996: exceptionalism crumbling, steps toward normalization. The forces driving the process of normalization are investigated. Since 1996: normalization, normality. The forms and perspectives of the changes made in the management of HIV and AIDS are elucidated using examples from the fields of health care, primary prevention and drug policies. AIDS health-policy innovations, their risks and opportunities in the course of normalization are investigated. Three possible paths of development are identified: stabilization, generalization and retreat. The chances of utilizing innovations developed in connection with AIDS for the modernization of health policy in other fields of prevention and patient care vary from country to country with the degree to which AIDS exceptionalism has been institutionalized and the distance of these innovations from medical, therapeutic events. PMID- 10795968 TI - Making sense of everyday pain. AB - A social constructionist analysis is reported of how sense is made of everyday pain. Q factor analysis is used within a critical framework as Q methodology. Sixty-one participants completed the procedure. Eight factors or accounts of everyday pain were derived. These are reported as pain as malfunction, pain as self-growth, pain as spiritual growth, pain as alien invasion, pain as coping and control, pain as abuse, pain as homeostatic mechanism and pain and power. Common to all of the accounts is the theme of how pain relates to self, and in particular, of whether pain can change self. This theme is expanded and discussed in terms of how self is protected and legitimated in a context of pain as a fundamental threat. Implications of this study for how to understand the experience of 'abnormal' pain are discussed, as are possible new research routes. PMID- 10795969 TI - Attitudes towards organ donation in Hong Kong. AB - In this paper, a survey is conducted to study the attitudes of Hong Kong residents towards organ donation after death. It is found that 60.3% of the respondents are willing to donate organs, which is comparable to those cited in the literature. The results also indicate that the willingness to donate is related to age, occupation and attitude. A factor analysis of attitudes yielded four factors, including form of burial, altruism, lack of understanding on organ donation and lack of confidence in professional conduct of medical doctors. Further analysis of the factor scores provides information that is useful to the design of effective public education programs in organ donation. PMID- 10795970 TI - Measurement of community beliefs about colorectal cancer. AB - Few educational campaigns have focused on bowel cancer, though studies have indicated that members of the community need and want current information about relevant issues. In order to facilitate research in this area, reliable and valid measures of community attitudes are needed. Content validity of a survey instrument was obtained through use of a Delphi process with Directors of Education from the Australia Cancer Council and focus group discussions with informed members of the public. The subsequent survey of community perceptions about colorectal cancer included a broad range of content areas related to the risk of bowel cancer, preventing and coping with bowel cancer and beliefs about susceptibility and severity. The construct validity of these content areas was investigated by use of a factor analysis and confirmation of an association with related predictor variables. Two measures related to personal influence and anticipated coping responses showed favourable psychometric properties, including moderate to high levels of internal consistency and test-retest reliability. A test of the concurrent validity of these measures requires further development of instruments related to colorectal cancer or adaptation of measures from other areas of health research. PMID- 10795971 TI - Gender comparisons of income expectations in the USA at the beginning of medical school during the past 28 years. AB - This study was designed to investigate gender differences in the USA, in anticipated professional income. Participants were 5314 medical students (3880 men, 1434 women) who entered Jefferson Medical College between 1970 and 1997. The annual peak professional income estimated at the beginning of medical school was the dependent variable and gender within selected time periods was the independent variable. Results showed significant differences between men and women on their anticipated future incomes in different time periods. Women generally expected 23% less income than men. The effect size estimates of the differences were moderately high. The gender gap in income expectations was more pronounced for those who planned to pursue surgery than their counterparts who planned to practice family medicine or pediatrics. A unique feature of this study is that its outcomes could not be confounded by active factors such as experience, working hours, age and productivity. Findings suggest that social learning may contribute to gender gap in anticipated income. PMID- 10795972 TI - The epidemiology of dementia in Africa: a review. AB - Very few of the 100 studies of the prevalence of dementia have been carried out in Africa. Much of the early work concerned small hospitalised samples. However, a series of studies from Ibadan, Nigeria, have produced consistently low rates for dementia, especially for Alzheimer's Disease. The most recent studies reveal rather higher rates, but still lower than surveys carried out elsewhere. The possible reasons for these findings are considered: differential survival rates, the hiding of cases by relatives because of stigma, reluctance to seek medical assistance as inappropriate, poor access to medical care, the feeling that the old person has come to the end of his useful life and defective case-finding techniques. The need for further research is emphasised. PMID- 10795973 TI - Marginal impact of psychosocial factors on multimorbidity: results of an explorative nested case-control study. AB - This study examines differences between subjects with zero, one or two or more new diseases in a period of three years, with regard to demographic characteristics, socioeconomic status, life style, medical family history and current diseases in the family, psychological and sociological characteristics. This was studied using a primary care based nested case-control study. Data were available from 3745 cases and controls, all aged 20 years and older. All subjects were sampled from the Registration Network Family Practices, which is a computerised continuous primary care database. Cases were defined as subjects with new multimorbidity (two or more new diseases) registered in a period of three years and two groups of controls were operationalised as subjects with either one or no new diseases registered in the same period. Determinants were assessed by means of a postal questionnaire. Increasing age, higher number of previous diseases and low socioeconomic status were strongly associated with both morbidity and multimorbidity. After adjustment for these basic variables, the occurrence of multimorbidity was more frequent among subjects who did not report (volunteer) work or study, who had an active coping style, a high occupational class and an external locus of control. Profiles for subjects at risk for morbidity and multimorbidity seem to differ. PMID- 10795974 TI - Implementation of FFT convolution and multigrid superposition models in the FOCUS RTP system. AB - In radiotherapy treatment planning, convolution/superposition algorithms currently represent the best practical approach for accurate photon dose calculation in heterogeneous tissues. In this work, the implementation, accuracy and performance of the FFT convolution (FFTC) and multigrid superposition (MGS) algorithms are presented. The FFTC and MGS models use the same 'TERMA' calculation and are commissioned using the same parameters. Both models use the same spectra, incorporate the same off-axis softening and base incident lateral fluence on the same measurements. In addition, corrections are explicitly applied to the polyenergetic and parallel kernel approximations, and electron contamination is modelled. Spectra generated by Monte Carlo (MC) modelling of treatment heads are used. Calculations using the MC spectra were in excellent agreement with measurements for many linear accelerator types. To speed up the calculations, a number of calculation techniques were implemented, including separate primary and scatter dose calculation, the FFT technique which assumes kernel invariance for the convolution calculation and a multigrid (MG) acceleration technique for the superposition calculation. Timing results show that the FFTC model is faster than MGS by a factor of 4 and 8 for small and large field sizes, respectively. Comparisons with measured data and BEAM MC results for a wide range of clinical beam setups show that (a) FFTC and MGS doses match measurements to better than 2% or 2 mm in homogeneous media; (b) MGS is more accurate than FFTC in lung phantoms where MGS doses are within 3% or 3 mm of BEAM results and (c) FFTC overestimates the dose in lung by a maximum of 9% compared to BEAM. PMID- 10795975 TI - Proton spectroscopic imaging of polyacrylamide gel dosimeters for absolute radiation dosimetry. AB - Proton spectroscopy has been evaluated as a method for quantifying radiation induced changes in polyacrylamide gel dosimeters. A calibration was first performed using BANG-type gel samples receiving uniform doses of 6 MV photons from 0 to 9 Gy in 1 Gy intervals. The peak integral of the acrylic protons belonging to acrylamide and methylenebisacrylamide normalized to the water signal was plotted against absorbed dose. Response was approximately linear within the range 0-7 Gy. A large gel phantom irradiated with three, coplanar 3 x 3 cm square fields to 5.74 Gy at isocentre was then imaged with an echo filter technique to map the distribution of monomers directly. The image, normalized to the water signal, was converted into an absolute dose map. At the isocentre the measured dose was 5.69 Gy (SD = 0.09) which was in good agreement with the planned dose. The measured dose distribution elsewhere in the sample shows greater errors. A T2 derived dose map demonstrated a better relative distribution but gave an overestimate of the dose at isocentre of 18%. The data indicate that MR measurements of monomer concentration can complement T2-based measurements and can be used to verify absolute dose. Compared with the more usual T2 measurements for assessing gel polymerization, monomer concentration analysis is less sensitive to parameters such as gel pH and temperature, which can cause ambiguous relaxation time measurements and erroneous absolute dose calculations. PMID- 10795976 TI - Optimized MR imaging for polyacrylamide gel dosimetry. AB - Polyacrylamide gels are a powerful tool to measure radiation dose by quantifying the NMR T2 relaxation times of the irradiated gel. The exploitation of these radiation sensitive gels in clinical radiotherapy requires accurate mapping of T2 values. This paper describes the optimization strategy used to identify accurate and practical methods of measuring the range of T2 values typical of gel dosimeters (140-700 ms). The MR imaging techniques used to measure T2 values and the choice of image acquisition parameters are described. Four sequences are compared and the results are analysed in terms of accuracy, signal-to-noise ratio and acquisition time. A multiple spin echo sequence was found to yield the most accurate results (98.9%). Single spin echo sequences, such as Hahn spin echo and EPI spin echo, were found to measure gel T2 values with an accuracy of 90.1%. This paper reports the importance of careful selection and optimization of the MR imaging sequences for accurate and reliable polyacrylamide gel dosimetry. PMID- 10795977 TI - An investigation of the chemical stability of a monomer/polymer gel dosimeter. AB - The aim of this work is to investigate the temporal stability of a polyacrylamide gelatin hydrogel used for 3D monomer/polymer gel dosimetry techniques involving different methods of analysis. Long-term instabilities for a similar gel have recently been reported, but differ markedly from those described in this work. Two kinds of long-term instabilities are described. One affects the slope of the dose-R2 plot and is related to post-irradiation polymerization of the comonomer/polymer aggregates. It is observed that post-irradiation polymerization only lasts 12 hours after irradiation. The other instability affects the intercept of the dose-R2 plot, lasts for up to 30 days and is related to the gelation process of gelatin. Further studies were performed on gelatin gels of varying compositions to obtain a better understanding of the molecular mechanism that causes the instability due to gelation. The studies included observations of the spin-spin and spin-lattice relaxation rates in combination with diffusion measurements and optical measurements. It is shown that the heating history during the manufacture of the gel affects the absolute R2 value of the gel but not its variation. The findings presented in this study may help in producing more stable and reproducible monomer/polymer gel dosimeters. PMID- 10795978 TI - Cavity theory applied to the dosimetry of systemic radiotherapy of bone metastases. AB - A two-component model of an osteoblastic metastatic lesion has been developed to determine the absorbed dose delivered to soft tissue during systemic radiotherapy of osseous metastases. Doses to soft tissue from radioisotopes distributed in bone were calculated using Burlin's general cavity theory. A correction term was used to account for the absence of charged particle equilibrium within the metastatic lesion. Radiation doses for 153Sm, 186Re, 89Sr and 32P were calculated for several physiologically realistic lesion structures. Burlin's cavity weighting factor was greatest for higher energy isotopes and it decreased as the soft tissue cavity size increased. The correction for the absence of charged particle equilibrium also decreased with soft tissue pathlength, but increased with average bone pathlengths. Doses to soft tissue cavities ranged from 0.1 to 0.2 Gy MBq(-1) d(-1) for 153Sm to 0.5 to 0.6 Gy MBq(-1) d(-1) for 32P. Using the factors calculated in this work, the dose to soft tissue cavities within bone metastases can be calculated when the dose to adjacent bone has been determined, perhaps by autoradiography or electron paramagnetic resonance dosimetry. The doses calculated with this more accurate model of bone metastases demonstrate errors of 20% to 50% in previous calculations of the average dose to homogeneous metastatic lesions. PMID- 10795979 TI - An empirical model for megavoltage x-ray output factors. AB - An empirical model of the factors that determine the central axis dose at 10 cm depth in water for 4 MV, 6 MV and 18 MV photon beams is presented. Backscattering from the variable collimators into the dose monitoring ionization chamber can cause a variation of -0.5% to +1.8% in the dose per monitor unit in accelerators with an electron facility. Forward emission towards the isocentre from the beam flattening filter and upper collimators is more dependent on the position of the upper variable collimator blades than the lower blades, so that they are not interchangeable in determining output factors, which can differ by up to 2%. The model includes the product of the monitor backscatter factor, normalized phantom scatter factor, normalized head scatter factor and inverse square law, corrected for the displacement of the virtual x-ray focus from the target. It can predict the dose to -/+0.83% for 4 MV, -/+0.80% for 6 MV photons and -/+0.82% for 18 MV photons. The normalized head scatter factor is a second-order polynomial of the modified equivalent square collimator, whose coefficients do not vary significantly with x-ray energy. The model was tested by comparison with independent measurements of output factor and generally agreed to around 1%. PMID- 10795980 TI - A neural network regression model for relative dose computation. AB - This paper describes a neural network (NN) regression model for relative dose computation. The input signal of the NN model includes depths and field sizes and the output is the relative dose, for example, percentage depth dose (%DD) or tissue-air ratio (TAR) in this paper. After a functional link has been created, the expressing ability of input patterns and the resolution of neural networks are enhanced. The trained neural network exhibits a good generalization and interpolation ability, and it can be easily extended to other relative dose regressions. We present two examples to verify the fitness of this model. The first calculates the percentage depth dose of a 14 MV x-ray beam for field sizes of 5 cm x 5 cm to 28 cm x 28 cm. The average error computed from the NN is less than 0.47% comparing with the original measured %DD. The second example calculates the TAR of a 4 MV x-ray beam for field sizes of 5 cm x 5 cm to 20 cm x 20 cm. In this example, the training data show that the average error computed from the NN is less than 0.48%, whereas that from the testing data is less than 0.37% after training. Such an NN model can be generalized to fit data for treatment planning with any linear accelerator and can also fit data stored as tissue-maximum ratio (TMR) and tissue-phantom ratio (TPR). PMID- 10795981 TI - Evaluation of the validity of a convolution method for incorporating tumour movement and set-up variations into the radiotherapy treatment planning system. AB - Modern radiotherapy techniques have developed to a point where the ability to conform to a particular tumour shape is limited by organ motion and set-up variations. The result is that dose distributions displayed by treatment planning systems based on static beam modelling are not representative of the dose received by the patient during a fractionated course of radiotherapy. The convolution-based method to account for these variations in radiation treatment planning systems has been suggested in previous work. The validity of the convolution method is tested by comparing the dose distribution obtained from this convolution method with the dose distribution obtained by summing the contribution to the total dose from each fraction of a fractionated treatment (for increasing numbers of fractions) and simulating random target position variations between fractions. For larger numbers of fractions (approximately or > 15) which are the norm for radical treatment schemes, it is clear that incorporation of movement by a convolution method could potentially produce a more accurate dose distribution. There are some limitations that have been identified, however, especially in relation to the heterogeneous nature of patient tissues, which require further investigation before the technique could be applied clinically. PMID- 10795982 TI - Computed tomography of x-ray index of refraction using the diffraction enhanced imaging method. AB - Diffraction enhanced imaging (DEI) is a new, synchrotron-based, x-ray radiography method that uses monochromatic, fan-shaped beams, with an analyser crystal positioned between the subject and the detector. The analyser allows the detection of only those x-rays transmitted by the subject that fall into the acceptance angle (central part of the rocking curve) of the monochromator/analyser system. As shown by Chapman et al, in addition to the x ray attenuation, the method provides information on the out-of-plane angular deviation of x-rays. New images result in which the image contrast depends on the x-ray index of refraction and on the yield of small-angle scattering, respectively. We implemented DEI in the tomography mode at the National Synchrotron Light Source using 22 keV x-rays, and imaged a cylindrical acrylic phantom that included oil-filled, slanted channels. The resulting 'refraction CT image' shows the pure image of the out-of-plane gradient of the x-ray index of refraction. No image artefacts were present, indicating that the CT projection data were a consistent set. The 'refraction CT image' signal is linear with the gradient of the refractive index, and its value is equal to that expected. The method, at the energy used or higher, has the potential for use in clinical radiography and in industry. PMID- 10795983 TI - Calculation of the neutron W value for neutron dosimetry below the MeV energy region. AB - The effective neutron W value for tissue-equivalent gas in the energy region from 5 keV to 5.7 MeV has been calculated using W values for recoil particles (protons, alpha particles, oxygen, carbon and nitrogen ions), which are produced by incident neutrons. The W value is assumed to be an energy-fluence-average over the W values of the recoil particles. The energy fluence spectra for the recoil particles are calculated by using a continuous slowing down approximation (CSDA). For the W values of recoil particles in the low-energy region, the recently evaluated data by Siebert et al and Taylor et al were used. Results are presented which show that the effective neutron W value depends strongly on energy in the low-energy region. This result indicates that neutron dose measurements using ionization chambers need a considerable correction of the W value in the low energy region. PMID- 10795984 TI - Polyvinyl alcohol-Fricke hydrogel and cryogel: two new gel dosimetry systems with low Fe3+ diffusion. AB - Two new Fricke dosimeter gel systems with low diffusion rates have been developed for 3D radiation dosimetry purposes. Both systems consist of a solution of 20% (by weight) polyvinyl alcohol (PVA) in a 50 mM H2SO4 solution with 0.4 mM ferrous ammonium sulphate and xylenol orange (FX). The difference in the two gels is the way that the gelation process was initiated: either by bringing the temperature to (a) +5 degrees C or (b) -20 degrees C before returning them to room temperature. These gels are termed 'hydrogel' and 'cryogel', respectively. The hydrogel is optically transparent, and can be used with either optical or MRI detection methods for dosimetric imaging. The cryogel is rubbery in texture but opaque, so its internal Fe3+ concentration can only be measured with MRI. The hydrogel's optical attenuation coefficient is linear (r2 = 0.99) with dose from 0 to 20 Gy with a sensitivity of 0.106 cm(-1) Gy(-1) (at 543 nm). In terms of MR relaxation rate, the dose response for both the hydrogel and cryogel was linear (r2 = 0.99) with a sensitivity of 0.020 s(-1) Gy(-1) (at 1.5 T). The Fe3+ diffusion coefficient (at 20 degrees C) was measured to be 0.14 mm2 h(-1), which is significantly lower than similar preparations reported for porcine gelatin or agarose. The PVA-FX gels can be stored for long periods of time before exposure to radiation, since the auto-oxidation rate was 10 times less than that of gelatin-Fricke recipes. The new gels developed in this work are a significant improvement on previous Fricke gel systems. PMID- 10795985 TI - Calibration of plane-parallel chambers and determination of p(wall) for the NACP and Roos chambers for 60Co gamma-ray beams. AB - Procedures for the calibration and use of plane-parallel ionization chambers in high-energy electron and photon beams have been given in the international code of practice IAEA TRS-381. In the present work, plane-parallel ionization chambers of the type PTW-34001 Roos and Scanditronix NACP02 have been calibrated using two N(K)-based procedures. For the NACP chamber the difference between the N(D,air) chamber factors determined in an electron beam and in a 60Co gamma-ray beam, respectively, is of the same magnitude as the experimental uncertainty. Results for the PTW Roos chambers, however, do not agree, in accordance with recent findings of other authors. The value determined in a 60Co gamma-ray beam is questioned and the reason for the discrepancy assigned to the correction factor for the perturbation due to the chamber wall, p(wall). New values of p(wall) have been experimentally determined by comparing absorbed dose measurements based on air-kerma and absorbed dose to water calibration procedures. A new p(wall) factor for the Roos chamber in 60Co gamma-ray beams in water (1.009+/-0.6%) was derived as the weighted average of the different determinations. The value is not significantly higher than the p(wall) factor given in TRS-381 (1.003+/-1.5%), but the combined standard uncertainty is reduced. The chamber to chamber variation for six commercial PTW Roos chambers and a Roos prototype was found to be very small. PMID- 10795986 TI - Converting absorbed dose to medium to absorbed dose to water for Monte Carlo based photon beam dose calculations. AB - Current clinical experience in radiation therapy is based upon dose computations that report the absorbed dose to water, even though the patient is not made of water but of many different types of tissue. While Monte Carlo dose calculation algorithms have the potential for higher dose accuracy, they usually transport particles in and compute the absorbed dose to the patient media such as soft tissue, lung or bone. Therefore, for dose calculation algorithm comparisons, or to report dose to water or tissue contained within a bone matrix for example, a method to convert dose to the medium to dose to water is required. This conversion has been developed here by applying Bragg-Gray cavity theory. The dose ratio for 6 and 18 MV photon beams was determined by computing the average stopping power ratio for the primary electron spectrum in the transport media. For soft tissue, the difference between dose to medium and dose to water is approximately 1.0%, while for cortical bone the dose difference exceeds 10%. The variation in the dose ratio as a function of depth and position in the field indicates that for photon beams a single correction factor can be used for each particular material throughout the field for a given photon beam energy. The only exception to this would be for the clinically non-relevant dose to air. Pre computed energy spectra for 60Co to 24 MV are used to compute the dose ratios for these photon beams and to determine an effective energy for evaluation of the dose ratio. PMID- 10795987 TI - EEG dipole source localization using artificial neural networks. AB - Localization of focal electrical activity in the brain using dipole source analysis of the electroencephalogram (EEG), is usually performed by iteratively determining the location and orientation of the dipole source, until optimal correspondence is reached between the dipole source and the measured potential distribution on the head. In this paper, we investigate the use of feed-forward layered artificial neural networks (ANNs) to replace the iterative localization procedure, in order to decrease the calculation time. The localization accuracy of the ANN approach is studied within spherical and realistic head models. Additionally, we investigate the robustness of both the iterative and the ANN approach by observing the influence on the localization error of both noise in the scalp potentials and scalp electrode mislocalizations. Finally, after choosing the ANN structure and size that provides a good trade off between low localization errors and short computation times, we compare the calculation times involved with both the iterative and ANN methods. An average localization error of about 3.5 mm is obtained for both spherical and realistic head models. Moreover, the ANN localization approach appears to be robust to noise and electrode mislocations. In comparison with the iterative localization, the ANN provides a major speed-up of dipole source localization. We conclude that an artificial neural network is a very suitable alternative for iterative dipole source localization in applications where large numbers of dipole localizations have to be performed, provided that an increase of the localization errors by a few millimetres is acceptable. PMID- 10795988 TI - Current densities in a 2 mm resolution anatomically realistic model of the body induced by low frequency electric fields. AB - Current density distributions in a fine resolution (2 mm) anatomically realistic voxel model of the human body have been calculated for uniform, low frequency vertically aligned electric fields for a body grounded and isolated from 50 Hz to 10 MHz. The voxel phantom NORMAN is used which has a height of 1.76 m and a mass of 73 kg. There are 8.3 million voxels in the body differentiated into 37 tissue types. Both finite-difference potential and time-domain methods were used. Results are presented for the current density averaged over 1 cm2 in muscle, heart, brain and retina. Electric field values required to reach the NRPB and ICNIRP basic restrictions on current density are derived and compared with the external field guidelines from these standards. PMID- 10795989 TI - Analysis of the distribution of intra-arterial microspheres in human liver following hepatic yttrium-90 microsphere therapy. AB - The microscopic distribution of microspheres in human liver following hepatic infusion of 32 microm diameter resin microspheres labelled with 90Y as treatment for an 80 millimetre diameter liver cancer has been investigated. Microspheres were found to deposit inhomogeneously in tissues, preferentially lodging in a region approximately 6 mm wide around the periphery of the tumour. A relative concentration of microspheres of 50 to 70 times that of normal hepatic parenchyma and 65 to 94 times that in the tumour centre was measured in this region. The deposition of spheres in the tumour periphery was not uniform, and cluster analysis showed that the spheres could be classified into clusters. The number of microspheres in a cluster was skewed towards low numbers and cluster sizes varied from 20 to 1500 microm. The observed deposition patterns indicate that the vascular tumour periphery will receive much greater radiation doses from radioactive microspheres than both normal tissue and the avascular tumour centre. PMID- 10795990 TI - Temperature simulations in tissue with a realistic computer generated vessel network. AB - The practical use of a discrete vessel thermal model for hyperthermia treatment planning requires a number of choices with respect to the unknown part of the patient's vasculature. This work presents a study of the thermal effects of blood flow in a simple tissue geometry with a detailed artificial vessel network. The simulations presented here demonstrate that an incomplete discrete description of the detailed network results in a better prediction of the temperature distribution than is obtained using the conventional bio-heatsink equation. Therefore, efforts to obtain information on the positions of the large vessels in an individual hyperthermia patient will be rewarded with a more accurate prediction of the temperature distribution. PMID- 10795991 TI - A comparison study of linear reconstruction techniques for diffuse optical tomographic imaging of absorption coefficient. AB - We compare, through simulations, the performance of four linear algorithms for diffuse optical tomographic reconstruction of the three-dimensional distribution of absorption coefficient within a highly scattering medium using the diffuse photon density wave approximation. The simulation geometry consisted of a coplanar array of sources and detectors at the boundary of a half-space medium. The forward solution matrix is both underdetermined, because we estimate many more absorption coefficient voxels than we have measurements, and ill conditioned, due to the ill-posedness of the inverse problem. We compare two algebraic techniques, ART and SIRT, and two subspace techniques, the truncated SVD and CG algorithms. We compare three-dimensional reconstructions with two dimensional reconstructions which assume all inhomogeneities are confined to a known horizontal slab, and we consider two 'object-based' error metrics in addition to mean square reconstruction error. We include a comparison using simulated data generated using a different FDFD method with the same inversion algorithms to indicate how our conclusions are affected in a somewhat more realistic scenario. Our results show that the subspace techniques are superior to the algebraic techniques in localization of inhomogeneities and estimation of their amplitude, that two-dimensional reconstructions are sensitive to underestimation of the object depth, and that an error measure based on a location parameter can be a useful complement to mean squared error. PMID- 10795992 TI - Investigation of a large-area phased array for focused ultrasound surgery through the skull. AB - Non-invasive treatment of brain disorders using ultrasound would require a transducer array that can propagate ultrasound through the skull and still produce sufficient acoustic pressure at a specific location within the brain. Additionally, the array must not cause excessive heating near the skull or in other regions of the brain. A hemisphere-shaped transducer is proposed which disperses the ultrasound over a large region of the skull. The large surface area covered allows maximum ultrasound gain while minimizing undesired heating. To test the feasibility of the transducer two virtual arrays are simulated by superposition of multiple measurements from an 11-element and a 40-element spherically concave test array. Each array is focused through an ex vivo human skull at four separate locations around the skull surface. The resultant ultrasound field is calculated by combining measurements taken with a polyvinylidene difluoride needle hydrophone providing the fields from a 44 element and a 160-element virtual array covering 88% and 33% of a hemisphere respectively. Measurements are repeated after the phase of each array element is adjusted to maximize the constructive interference at the transducer's geometric focus. An investigation of mechanical and electronic beam steering through the skull is also performed with the 160-element virtual array, phasing it such that the focus of the transducer is located 14 mm from the geometric centre. Results indicate the feasibility of focusing and beam steering through the skull using an array spread over a large surface area. Further, it is demonstrated that beam steering through the skull is plausible. PMID- 10795993 TI - The potential use of polymer gel dosimetry in boron neutron capture therapy. AB - Polymer gels with and without 60 ppm of 10B were exposed to an epithermal neutron beam produced by the Dynamitron at the University of Birmingham on two separate occasions. Eight vials containing the gel, four with and four without boron, were irradiated in pairs in a water phantom for 5 h. The maximum dose was calculated to be 9 Gy in A-150 tissue equivalent plastic, 4 cm deep in the phantom. Measurements were made of the variation of relaxation rates of the gels with depth in a phantom. These were compared with calculations using the MCNP Monte Carlo program and the gel response followed the general trend of the results of the calculations. The calculations showed that the absence of boron gave 66.1% and 44.3% of the absorbed dose with boron and the measurements showed the response of the gel without boron to give 65+/-2% and 41+/-6% of the response with boron for the two halves of the first vial. All the gel measurements showed an enhancement in absorbed dose when boron was added. These results indicate that polymer gels may have a role in measuring the enhancement of absorbed dose due to boron in an epithermal or thermal neutron. PMID- 10795994 TI - Effects of cellular repair and proliferation on targeted radionuclide therapy: a modelling study. AB - A previous targeted radionuclide therapy modelling study has been extended to include the radiobiological effects of cellular repair and proliferation. Dose distributions have been converted into biologically effective dose (BED) distributions using a previously published formulation. With suitable estimated parameters, corrected tumour control probability (TCP) values were derived. The dependence of BED on the physical half-life of the radionuclide was also modelled. Results indicate that the TCP is greater when a shorter physical half life is employed. PMID- 10795995 TI - A quality control method for detecting energy changes of medical accelerators. AB - A description is presented of a simple and sensitive method for detecting a change in the energy of the electrons bombarding the target of medical accelerators. This technique is useful for x-ray beams with end point energy in the range of 15.7 to 25 MeV. The method is based on the photoactivation of 160 and 14N in a small sample of ammonium nitrate. It was found that the ratio of the activity induced in the oxygen divided by that produced in the nitrogen can be used as a quality control technique to detect a change in the energy of the electrons that bombard the target of the accelerator. An electron energy change of the order of 0.2 MeV can be determined using this method. PMID- 10795996 TI - Calibration of a tomotherapeutic MVCT system. AB - Megavoltage CT provides the ability to image the patient before, during or after a radiotherapy treatment. This allows one to verify not only the placement of a patient's external boundary, but also the locations of internal anatomy. In addition, the reconstructed MVCT values are potentially useful for treatment planning inhomogeneity corrections and dose reconstruction. To this end, dosimetric calibration of the University of Wisconsin Tomotherapy Benchtop MVCT system was investigated. It was found that MVCT values correlate extremely well with electron density and that unlike kilovoltage CT, this correlation is well maintained for higher atomic number materials. Improvements of the order of 1% in the dosimetric calculations of high atomic number materials should be possible by deriving input images from MVCT as opposed to kVCT, and calibrating in terms of electron density, as opposed to physical density. PMID- 10795998 TI - Effects of dipyridamole on left ventricular function. AB - BACKGROUND: Changes induced by dipyridamole infusion on left ventricular function in healthy individuals have not been investigated by gated myocardial perfusion single photon emission computed tomographic (SPECT) imaging. METHODS AND RESULTS: This study examined the amplitude and duration of changes induced by dipyridamole infusion on left ventricular function as assessed by technetium 99m sestamibi gated SPECT in 18 subjects with a low likelihood of coronary artery disease. Twenty mCi (740 MBq) of Tc-99m sestamibi were injected at rest. Three different consecutive gated SPECT images were performed 60 minutes later: baseline at rest, during the infusion of 0.76 mg/kg of dipyridamole, and 1 hour later. No patient received aminophylline. Left ventricular ejection fraction (LVEF), end-diastolic volume, and end-systolic volume were automically computed. Heart rate (HR) and blood pressure were regularly monitored. Mean LVEF was 63.2% +/- 8.0% baseline at rest, increased to 73.8% +/- 8.2% (P = .0001) during dipyridamole infusion, and returned to baseline values (63.0% +/- 7.5%) 1 hour later. End-diastolic volume did not vary significantly, and end-systolic volume decreased (from 32.2 +/- 19.5 to 26.6 +/- 17.9 u, P = .002) and returned to baseline values (32.7 +/- 15.6 u) 1 hour later. Dipyridamole induced moderate HR acceleration (from 80.2 +/- 15.0 to 96.5 +/- 9.6 beats/min, P < .001) and a slight decrease in diastolic blood pressure (from 80.6 +/- 8.1 to 70.1 +/- 9.0 mm Hg, P < .001). However, 1 hour later, HR and blood pressure had returned to baseline values. CONCLUSIONS: Dipyridamole increases LVEF and HR and decreases diastolic blood pressure slightly in healthy individuals. Because dipyridamole gated SPECT imaging acquisition is usually started 60 minutes after dipyridamole infusion, LVEF is in fact estimated at rest. PMID- 10795997 TI - Effect of L-arginine administration on myocardial thallium-201 perfusion during exercise in patients with angina pectoris and normal coronary angiograms. AB - OBJECTIVES: We tested the hypothesis that an exogenous supplement of L-arginine could alleviate coronary perfusion abnormality during exercise in patients with angina pectoris and normal coronary arteries. METHODS AND RESULTS: Twelve patients underwent exercise thallium-201 scintigraphy without medication (control) and after intravenous administration of L-arginine. Exercise time was prolonged in the L-arginine study compared with the control (482 s vs 540 s, P < .05). TI-201 extent score was improved in the L-arginine study (0.33 vs 0.26, P < .05), and the severity score was also improved (23.7 vs 16.9, P < .05). In 7 of the 12 patients whose TI-201 redistribution disappeared in the L-arginine study, the percent increase in serum L-citrulline concentration during exercise was larger than that of the remaining 5 patients (18% vs 0.9%, P < .01). The percent reduction in epicardial coronary diameter in response to acetylcholine was also greater in the former group (28.3% vs 11.1%, P < .05). CONCLUSION: Exogenous L arginine improved myocardial perfusion during exercise in a subset of patients with angina pectoris and normal coronary arteries, probably by increasing production of nitric oxide. PMID- 10795999 TI - Evaluation of myocardial infarct size in rat heart by pinhole SPECT. AB - BACKGROUND: High-resolution single photon emission computed tomography (SPECT) with a pinhole collimator is a new method for evaluating the regional properties of radiopharmaceuticals in small laboratory animals in vivo. Although several reports of normal images of rat taken by this new technique are available, there are as yet few reports on its use in disease models, such as myocardial infarction. In this study, we clearly visualized myocardial flow in the rat heart with myocardial infarction using this system, and evaluated the relationship between SPECT images and histologic analysis. METHODS AND RESULTS: For visualization of myocardial flow in rat heart, 201Tl images were taken just before and 24 days after left coronary artery ligation. The images were taken using a 4-head SPECT scanner with pinhole collimators. The percent infarct size on 201Tl-SPECT imaging (%SI) and the defect score were then assessed and compared with the percent infarct size on histologic analysis (%HI). Both the %SI and defect score correlated well with %HI (r = 0.97 and 0.74, respectively). CONCLUSION: Serial SPECT imaging using pinhole collimators permits estimates of myocardial flow even in small laboratory animals noninvasively in vivo. PMID- 10796000 TI - Acute ischemic dysfunction alters coronary flow reserve in remote nonischemic regions: potential mechanical etiology identified in an acute canine model. AB - BACKGROUND: Impaired coronary flow reserve (CFR) has been observed in remote nonischemic regions in patients after myocardial infarction. The mechanism for this impairment in remote nonischemic CFR remains undefined. This study evaluates the effect of progressive regional ischemic dysfunction on function in remote nonischemic regions, and the effect of the extent of dysfunction on remote nonischemic coronary flow and CFR. METHODS: In an anesthetized open-chest canine model (n = 7) of acute progressive distal and proximal left anterior descending (LAD) coronary artery occlusion, regional myocardial thickening fraction and coronary flow and CFR were measured with Doppler probes. CFR was assessed by an intracoronary injection of 36 microg of adenosine. Changes in thickening fraction and CFR were evaluated for isovolumic, ejection, and diastolic phases. Changes in resting regional flow were also assessed using radiolabeled microspheres. The extent of the ischemic area was defined as regions of myocardium with endocardial microsphere blood flow less than 0.3 mL/min/g. RESULTS: The ischemic area increased from 12% +/- 1% of left ventricle with distal occlusion to 30% +/- 2% of left ventricle with proximal occlusion (P < .001). The LAD thickening fraction decreased significantly from baseline (18% +/- 1%) to distal (-8% +/- 1%,) and proximal (-4% +/- 1%) occlusion (P < .001 for distal and proximal vs baseline). Isovolumic bulging in the LAD region was associated with a progressive increase in thickening fraction in the remote nonischemic left circumflex (LCX) artery region (baseline 12% +/- 1%; distal occlusion 15% +/- 2%, P = .014 vs baseline; proximal occlusion 17% +/- 2%, P = .02 vs baseline). Most of the increase in remote thickening fraction occurred during the isovolumic phase. There was no significant change in resting flow in remote nonischemic LCX regions or global hemodynamic parameters. However, there was a progressive decrease in remote nonischemic CFR (baseline 2.44 +/- 0.3), distal occlusion (2.19 +/- 0.31; P = .055 vs baseline), and proximal occlusion (1.79 +/- 0.22; P = 0.004 vs baseline, and P = .012 vs distal occlusion). A progressive decrease in CFR was noted in each phase of the cardiac cycle. CONCLUSION: In a canine model of acute progressive distal and proximal coronary occlusion, we observed a progressive decrease in CFR in remote nonischemic regions concurrent with an increase in the extent of ischemia. The decrease in remote nonischemic CFR was associated with ischemia-induced isovolumic bulging, which placed the remote regions at a mechanical disadvantage. These observations suggest a potential mechanical etiology for the observed impairment in remote CFR. Alterations in remote nonischemic CFR during acute ischemia may have important clinical implications for perfusion scintigraphy. PMID- 10796001 TI - Optimal timing for initial and redistribution technetium 99m-N-NOET image acquisition. AB - BACKGROUND: Bis (N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium-99m (V) (Tc-99m-N-NOET) is a new Tc-99m-labeled myocardial perfusion imaging agent that redistributes. We sought to determine the optimal timing for acquiring initial and delayed images to maximize sensitivity for the detection of coronary stenoses. METHODS: Twelve anesthetized dogs with critical stenoses of the left anterior descending coronary artery were infused with adenosine (250 microg/kg/min) or MRE-0470, an adenosine A2a agonist (0.6 microg/kg/min x 10 minutes), and Tc-99m-N-NOET (8 mCi; 296 MBq) was injected intravenously at peak flow. Myocardial and lung Tc-99m-N-NOET activities were determined by serial quantitative imaging and arterial blood sampling was performed over 2 hours. RESULTS: Left anterior descending/left circumflex artery defect count ratios showed rapid redistribution during the first 10 minutes after Tc-99m-N-NOET injection (0.66 +/- 0.02 at 2 minutes to 0.73 +/- 0.01 at 10 minutes; P < .01). Redistribution was nearly complete by 120 minutes (defect ratio = 0.87 +/- 0.03; P < or = .01 vs 2 minutes). Lung activity fell significantly during the first 10 minutes from a heart/lung activity ratio of 1.07 +/- 0.05 (2 minutes) to 1.44 +/- 0.09 (10 minutes; P < or = .01). CONCLUSION: Initial stress Tc-99m-N-NOET images should be acquired within 10 minutes after injection, whereas delayed images can be obtained as early as 2 hours later. Lung activity clears rapidly, permitting acquisition of good-quality poststress cardiac images. These Tc-99m-N-NOET uptake and redistribution kinetics after vasodilator stress provide important information for designing clinical imaging protocols for optimal identification of inducible ischemia. PMID- 10796003 TI - Comparative localization of myocardial ischemia by exercise electrocardiography and myocardial perfusion SPECT. AB - BACKGROUND: Prior angiographic study has shown that the patterns of ST-segment depression during exercise do not provide localizing information of the responsible coronary lesion. However, little is known regarding the ability of exercise-induced ST-segment displacement to localize myocardial perfusion defects. METHODS AND RESULTS: We studied 552 consecutive patients without prior myocardial infarction who had reversible perfusion defect in one vascular territory on rest 201Tl/exercise 99mTc-labeled sestamibi dual-isotope myocardial perfusion single photon emission computed tomography (SPECT) and ischemic ST depression or elevation during exercise. Of these, 192 patients had angiographically documented coronary artery disease (CAD). Two hundred thirty-two patients had maximal ST depression in anterior leads, 247 patients had maximal ST depression in inferior leads, and 45 patients had similar maximal ST depression in both anterior and inferior leads. Twenty-eight (5%) patients had ST elevation with absent Q waves. In patients with maximal ST depression in anterior leads, perfusion defects were found in the territory of the left anterior descending coronary artery (LAD) in 30%, in the territory of the right coronary artery (RCA) in 52%, and in the territory of the left circumflex coronary artery (LCX) in 18%. In patients with maximal ST depression in inferior leads, perfusion defects were found in RCA territory in 44%, in the LAD territory in 42%, and in the LCX territory in 14%. Compared with exercise ST depression, the less common finding of ST elevation did provide accurate localization of perfusion defects. When ST elevation was greatest in the anterior leads, 96% of patients had LAD territory defects. When ST elevation was most prominent in the inferior leads, 100% patients had RCA territory defects. Data of coronary angiograms demonstrated that myocardial perfusion SPECT correctly identified the most stenotic coronary disease for LAD (94%), LCX (72%), and RCA (75%). CONCLUSIONS: The findings of this study indicate that the site of maximal ST-segment depression does not identify the localization of myocardial perfusion defects. However, the less common finding of exercise-induced ST-segment elevation does predict localization of myocardial ischemia. PMID- 10796002 TI - Burden of myocardial damage in cardiac allograft rejection: scintigraphic evidence of myocardial injury and histologic evidence of myocyte necrosis and apoptosis. AB - BACKGROUND: Because myocardial damage determines morbidity and outcomes in heart transplant rejection, assessment of total burden of myocardial damage is highly desirable. In addition to myocyte necrosis, programmed cell death, or apoptosis, has recently been shown to contribute to cardiac allograft rejection. In the present study, we noninvasively determined myocardial damage by antimyosin scintigraphy and compared it with necrotic and apoptotic myocardial damage in endomyocardial biopsy (EMB) specimens. METHODS AND RESULTS: Forty scintigraphic and histologic studies were simultaneously performed. Of these, 19 patients had no EMB evidence of allograft rejection (group I, International Society of Heart and Lung Transplantation [ISHLT] grade 0/4), 12 had mild rejection (group II, ISHLT grades 1A and 1B), and 9 had evidence of moderate allograft rejection (group III, ISHLT grades 2, 3A, and 3B). None of the biopsies demonstrated severe allograft rejection (ISHLT grade 4/4). The severity of global myocyte damage in 40 patients was assessed by antimyosin scintigraphy. Endomyocardial biopsies were performed in these patients within 48 hours of imaging study; biopsy specimens were characterized for presence of myocyte necrosis and apoptosis. Evidence of myocyte necrosis was observed in 9 (23%) of 40 EMB specimens. Nineteen EMB specimens of group I had no inflammation and no myocyte necrosis, 12 of group II specimens showed interstitial mononuclear cell infiltration (only) but no myocyte necrosis, and all 9 of group III specimens had evidence of cellular infiltration and myocyte damage. Myocyte necrosis was assessed by hematoxylin-eosin and trichrome staining of EMB specimens. On the other hand, apoptosis of myocytes, as assessed by TUNEL staining of DNA fragments, was seen in 22 (55%) of the 40 biopsy specimens: 47%, 58%, and 67% in groups I, II and III, respectively. Abnormal antimyosin scan findings, indicating presence of myocardial damage, were observed in 9 of the 19 patients in group I and in all patients in groups II and III. Although positive antimyosin scan results in group III patients are concordant with the presence of histologic myocardial necrosis, myocardial uptake of antimyosin antibodies in groups I and II (no apparent myocyte damage at light microscopic examination) could reflect either sampling error of the biopsy or ongoing apoptotic myocyte damage. CONCLUSIONS: Apoptosis of myocytes is frequently observed during cardiac allograft rejection. The presence of apoptotic myocytes in the absence of histologic rejection activity in patients with antimyosin uptake suggests that apoptosis could be an additional mechanism of transplant-associated myocardial damage. PMID- 10796004 TI - Statistical analysis of medical data. Part I: Univariable analysis. PMID- 10796005 TI - Revascularization strategies in patients with diabetes: evolving concepts. PMID- 10796007 TI - Technologists as teachers. PMID- 10796006 TI - Molecular genetics of cardiomyopathies. PMID- 10796008 TI - Small is beautiful: specialty imaging devices and the growth of nuclear cardiology. PMID- 10796009 TI - The role of myocardial perfusion imaging in vascular endothelial dysfunction. PMID- 10796010 TI - Technetium 99m-N-NOET: although not equivalent to thallium-201, it still offers new opportunities. PMID- 10796012 TI - Nuclear cardiology in the literature. PMID- 10796011 TI - Acute chest pain imaging in the outpatient setting. PMID- 10796013 TI - On routine stress perfusion studies: how do you grade defect size? PMID- 10796014 TI - Symbiotic induction of pyruvate dehydrogenase genes from Sinorhizobium meliloti. AB - Genes coding for components of the pyruvate dehydrogenase (PDH) multienzyme complex (PDHc) from Sinorhizobium meliloti, the alfalfa symbiont, have been isolated on the basis of their high expression in symbiotic bacteria. The Elp component, PDH, is encoded by two genes, pdhAalpha (1,047 bp) and pdhAbeta (1,383 bp), a situation encountered in the alpha-proteobacteria Rickettsia prowazekii and Zymomonas mobilis as well as in some gram-positive bacteria and in mitochondria. pdhAalpha and pdhAbeta precede pdhB (1,344 bp), which encodes the E2p component, dihydrolipoamide acetyltransferase, of the PDHc. No gene encoding the E3 component, lipoamide dehydrogenase, was found in the immediate vicinity of pdhA and pdhB genes. pdhAalpha, pdhAbeta and pdhB likely constitute an operon. Here, we provide evidence that pdhA expression is induced in the symbiotic stage, compared with free-living conditions. We demonstrate that symbiotic expression of pdhA genes does not depend on the fix LJ regulatory cascade that regulates nitrogen fixation and respiration gene expression in symbiotic S. meliloti cells. Induction of pdhA expression could be obtained under free-living conditions upon the addition of pyruvate to the culture medium. Induction by pyruvate and symbiotic activation of pdh gene expression take place at the same promoter. PMID- 10796015 TI - Evidence that the cAMP pathway controls emergence of both primary and appressorial germ tubes of barley powdery mildew. AB - Development of conidia of barley powdery mildew involves the formation of a primary germ tube (PGT), an appressorial germ tube (AGT), and an appressorium. Previously, it was found that cyclic AMP (cAMP) was involved in these developmental processes. Comparison of development on the host surface with two types of cellulose membrane revealed that frequency of PGT emergence was surface independent. On one type of cellulose, where the frequencies of both AGT and appressorial differentiation were similar to that on the host surface, cAMP levels and protein kinase A (PKA) activities had a biphasic pattern with peaks at 15 min and 4 h after inoculation (prior to PGT and AGT emergence, respectively). The effect of manipulating cAMP levels was tested on another type of cellulose membrane, which stimulated a lower degree of AGT and appressorial formation than the host surface. Cholera toxin and forskolin, activators of adenylyl cyclase, significantly increased PGT emergence, but cAMP did not. Cholera toxin, forskolin, and cAMP increased the frequency of AGT and appressorial formation, but in a time-dependent manner. PMID- 10796016 TI - Arabidopsis thaliana EDS4 contributes to salicylic acid (SA)-dependent expression of defense responses: evidence for inhibition of jasmonic acid signaling by SA. AB - The Arabidopsis enhanced disease susceptibility 4 (eds4) mutation causes enhanced susceptibility to infection by the bacterial pathogen Pseudomonas syringae pv. maculicola ES4326 (Psm ES4326). Gene-for-gene resistance to bacteria carrying the avirulence gene avrRpt2 is not significantly affected by eds4. Plants homozygous for eds4 exhibit reduced expression of the pathogenesis-related gene PR-1 after infection by Psm ES4326, weakened responses to treatment with the signal molecule salicylic acid (SA), impairment of the systemic acquired resistance response, and reduced accumulation of SA after infection with Psm ES4326. These phenotypes indicate that EDS4 plays a role in SA-dependent signaling. SA has been shown to have a negative effect on activation of gene expression by the signal molecule jasmonic acid (JA). Two mutations that cause reduced SA levels, eds4 and pad4, cause heightened responses to inducers of JA-dependent gene expression, providing genetic evidence to support the idea that SA interferes with JA-dependent signaling. Two possible working models of the role of EDS4 in governing activation of defense responses are presented. PMID- 10796017 TI - Host suppressors in Arabidopsis thaliana of mutations in the movement protein gene of Cauliflower mosaic virus. AB - A novel genetic screen was used to identify host factors in Arabidopsis thaliana that suppress mutations in the Cauliflower mosaic virus (CaMV) movement protein gene (gene I). A series of small mutations was made in gene I and the mutations were tested for their suitability in a suppressor screen. The first round of screening yielded only revertants or second-site mutations in gene I. A derivative of one of the second-site mutant viruses (N7) that was delayed in symptom production was used in a second round of screening for suppressor plants that accelerated symptom production. Two candidate suppressor plants were found that accelerated by 1 to 4 days the first appearance of symptoms caused by the mutant viruses. One of the suppressors (5-2), called asc1 (acceleration of symptoms by CaMV N7), was mapped to chromosome 1. Two additional loci that differentially affect N7 virus susceptibility in the parental Columbia and Ler ecotypes were mapped to chromosomes 3 and 4 by quantitative trait locus (QTL) analysis. PMID- 10796018 TI - P42 movement protein of Beet necrotic yellow vein virus is targeted by the movement proteins P13 and P15 to punctate bodies associated with plasmodesmata. AB - Cell-to-cell movement of Beet necrotic yellow vein virus (BNYVV) is driven by a set of three movement proteins--P42, P13, and P15--organized into a triple gene block (TGB) on viral RNA 2. The first TGB protein, P42, has been fused to the green fluorescent protein (GFP) and fusion proteins between P42 and GFP were expressed from a BNYVV RNA 3-based replicon during virus infection. GFP-P42, in which the GFP was fused to the P42 N terminus, could drive viral cell-to-cell movement when the copy of the P42 gene on RNA 2 was disabled but the C-terminal fusion P42-GFP could not. Confocal microscopy of epidermal cells of Chenopodium quinoa near the leading edge of the infection revealed that GFP-P42 localized to punctate bodies apposed to the cell wall whereas free GFP, expressed from the replicon, was distributed uniformly throughout the cytoplasm. The punctate bodies sometimes appeared to traverse the cell wall or to form pairs of disconnected bodies on each side. The punctate bodies co-localized with callose, indicating that they are associated with plasmodesmata-rich regions such as pit fields. Point mutations in P42 that inhibited its ability to drive cell-to-cell movement also inhibited GFP-P42 punctate body formation. GFP-P42 punctate body formation was dependent on expression of P13 and P15 during the infection, indicating that these proteins act together or sequentially to localize P42 to the plasmodesmata. PMID- 10796019 TI - Regulation of tomato leaf curl viral gene expression in host tissues. AB - The regulation of expression of the two virion-sense (V1 and V2) and four complementary-sense (C1, C2, C3, and C4) open reading frames (ORFs) of Tomato leaf curl virus (TLCV) was studied in both stably and transiently transformed Nicotiana tabacum tissues with fusions with the beta-glucuronidase (GUS) reporter gene. GUS-expressing transgenic lines were obtained with each of the four complementary-sense gene-GUS fusion constructs and with truncated versions of the virion-sense gene-GUS fusion constructs (V1GUSdeltaC and V2GUSdeltaC) lacking complementary-sense sequences encoding the C1, C2, and C3 ORFs. However, little or no GUS expression was observed in kanamycin-resistant plants transformed with full-length, virion-sense gene constructs (V1GUS and V2GUS) constituting the complete viral genome. In contrast, V1GUS and V2GUS were found to direct high level GUS expression in transient assays with tobacco protoplasts, suggesting that integration of viral constructs containing functional, complementary-sense genes may lead to repression or deletion of the introduced constructs in transgenic tissues. V2GUS expression in the transient protoplast assay was found to be severely curtailed by specific mutation of the C2 ORF, supporting a role for the C2 protein in transactivation of TLCV virion-sense gene expression. TLCV ORF-GUS constructs displayed distinctive tissue expression patterns in transgenic tobacco plants that could be divided into constitutive (C1, C4, and V2GUSdeltaC), predominantly vascular (C2, C3), or reduced expression in cells associated with the vascular bundles (V1GUSdeltaC). The significance of these results is discussed in terms of current models of gene function and regulation in geminiviruses. PMID- 10796020 TI - Characterization of SNP1, a cell wall-degrading trypsin, produced during infection by Stagonospora nodorum. AB - Stagonospora (= Septoria) nodorum when grown in liquid culture with wheat cell walls as the sole carbon and nitrogen source secretes numerous extracellular depolymerases, including a rapidly produced, alkaline, trypsin-like protease (SNP1). The enzyme was purified 417-fold by cation exchange chromatography and has a molecular mass of 25 kDa on sodium dodecyl sulfate gels, pI 8.7, and pH optimum of 8.5. It cleaved peptide bonds on the carboxyl side of lysine or arginine, was strongly inhibited by the trypsin inhibitors aprotinin and leupeptin and weakly by phenylmethylsulfonyl fluoride, and its activity was stimulated by calcium. SNP1 has the characteristic, conserved, fungal, trypsin N terminus. Polymerase chain reaction (PCR) primers based on this sequence and the conserved trypsin active site were used to amplify a DNA fragment that facilitated isolation of the corresponding genomic clone from a lambda library of S. nodorum. The full-length sequence confirmed its identity as a trypsin-like protease containing the N-terminal sequence of the previously purified enzyme. Infected leaf tissue contained a protease, not present in controls, that coeluted with the fungal trypsin from cation exchange, and had properties (pI and inhibitor characteristics) similar to those of the fungal trypsin. SNP1 expression in planta was detected by Northern (RNA) blotting, reverse transcription PCR, and green fluorescent protein confocal microscopy. SNP1 released hydroxyproline from wheat cell walls. The release of hydroxyproline, together with its early expression in planta, suggests that SNP1 participates in the degradation of host cell walls during infection. PMID- 10796021 TI - Constitutive accumulation of a resveratrol-glucoside in transgenic alfalfa increases resistance to Phoma medicaginis. AB - Alfalfa (Medicago sativa) was transformed with a peanut (Arachis hypogaea) cDNA encoding resveratrol synthase (RS) transcriptionally regulated by an enhanced Cauliflower mosaic virus (CaMV) 35S promoter. Transgenic plants accumulated a new compound, not present in wild-type or vector-transformed alfalfa, that was identified as trans-resveratrol-3-O-beta-D-glucopyranoside (RGluc) by high pressure liquid chromatography (HPLC), UV, 1H- and 13C-nuclear magnetic resonance (NMR) analyses. RGluc concentration was highest in the youngest leaves (>15 microg per g fresh weight) and oldest stem internode segments (>10 microg per g fresh weight) while roots contained only trace amounts (<0.2 microg per g fresh weight). RS transcript levels were highest in leaves and stems, with comparatively little transcript accumulation in the roots, while an inverse pattern was observed for chalcone synthase (CHS) transcript levels. CHS directly competes with RS for the metabolic precursors p-coumaroyl CoA and malonyl CoA, and may also contribute to the developmental variations in RGluc levels by limiting the availability of substrates. Agar-plate bioassays indicated that both RGluc and resveratrol greatly inhibit hyphal growth of the alfalfa fungal pathogen Phoma medicaginis. Subsequently, RGluc-containing leaves were wound inoculated and showed a significant reduction (relative to control leaves) in the size of necrotic lesions, intensity of adjacent chlorosis, and number of fungal reproductive structures (pycnidia). Decreasing sporulation of this pathogen may greatly reduce disease spread and severity throughout the field. PMID- 10796022 TI - Decreased inositol 1,4,5-trisphosphate content in pathogen-challenged soybean cells. AB - Phosphoinositide-specific phospholipase C (PI-PLC) has been shown to be transiently activated when plant cells were treated with elicitors. We thus investigated the activity of PI-PLC when soybean cells were infected with the bacterial pathogen Pseudomonas syringae pv. glycinea, by measuring cellular cytosolic inositol 1,4,5-trisphosphate (IP3) levels. We observed that IP3 content decreased in both compatible and incompatible interactions. In vitro phosphatase activities were similar in both water control and infected cells with slightly lower IP3 degradation observed for infected cells, indicating that the reduced IP3 content in infected cells most likely results from reduced PI-PLC activity. We hypothesize that reduced IP3 content following infection may lead to suppression of various housekeeping activities of the cells, thus diverting the cellular resources either to the synthesis of defense-related compounds against pathogens, and/or to the growth of pathogens. PMID- 10796023 TI - avrPto enhances growth and necrosis caused by Pseudomonas syringae pv.tomato in tomato lines lacking either Pto or Prf. AB - AvrPto was introduced into three tomato genotypes with two biotic agents to study its role in compatible interactions. avrPto enhanced the capacity of the Pseudomonas syringae pv. tomato strain T1 to induce necrotic symptoms on tomato plants that lacked either Pto or Prf genes. The enhanced necrosis correlated with a small increase in bacterial growth. In planta expression of avrPto in isolation did not elicit necrosis in the absence of a functional Prf gene. PMID- 10796024 TI - Multiresistance genes of Rhizobium etli CFN42. AB - Multidrug efflux pumps of bacteria are involved in the resistance to various antibiotics and toxic compounds. In Rhizobium etli, a mutualistic symbiont of Phaseolus vulgaris (bean), genes resembling multidrug efflux pump genes were identified and designated rmrA and rmrB. rmrA was obtained after the screening of transposon-generated fusions that are inducible by bean-root released flavonoids. The predicted gene products of rmrAB shared significant homology to membrane fusion and major facilitator proteins, respectively. Mutants of rmrA formed on average 40% less nodules in bean, while mutants of rmrA and rmrB had enhanced sensitivity to phytoalexins, flavonoids, and salicylic acid, compared with the wild-type strain. Multidrug resistance genes emrAB from Escherichia coli complemented an rmrA mutant from R. etli for resistance to high concentrations of naringenin. PMID- 10796025 TI - Reaction-diffusion waves of actin filament polymerization/depolymerization in Dictyostelium pseudopodium extension and cell locomotion. AB - Cell surface movements and the intracellular spatial patterns and dynamics of actin filament (F-actin) were investigated in living and formalin-fixed cells of Dictyostelium discoideum by confocal microscopy. Excitation waves of F-actin assembly developed and propagated several micrometers at up to 26 microm/min in cells which had been intracellularly loaded with fluorescently labeled actin monomer. Wave propagation and extinction corresponded with the initiation and attenuation of pseudopodium extension and cell advance, respectively. The identification of chemical waves was supported by the ring, sphere, spiral and scroll wave patterns, which were observed in the extensions of fixed cells stained with phalloidin-rhodamine, and by the similar, asymmetrical [F-actin] distribution in wavefronts in living and fixed cells. These F-actin patterns and dynamics in Dictyostelium provide evidence for a new supramolecular state of actin, which propagates as a self-organized, reaction-diffusion wave of reversible F-actin assembly and affects pseudopodium extension. Actin's properties of oscillation and self-organization might also fundamentally determine the nature of the eukaryotic cell's reactions of adaptation, timing and signal response. PMID- 10796026 TI - Surface properties of aqueous solutions of L-leucine. AB - The surface tension, sigma, of solutions of L-leucine (CH3)2CHCH2CH(NH2)COOH in water, as well as in aqueous solutions of NaOH and HCl were measured in the temperature range between 278 and 308 K using the Wilhelmy plate method. L Leucine was found to be a very weak surfactant, which can be understood if assuming strong interactions of this solute with the water structure. Striking differences were observed in the surface entropy of L-leucine solutions in water, 0.5 M HCl and 0.5 M NaOH. Moreover, surface activity of the solute is much lower than that supposed taking into account the hydrophobicity of this amino acid. It was concluded that the observed phenomena are caused by the water structure changes close to the side chain of leucine, caused by enforced hydrophobic hydration, i.e. formation of clathrate-like hydrates. PMID- 10796027 TI - Calculation of the standard molal thermodynamic properties of aqueous biomolecules at elevated temperatures and pressures II. Unfolded proteins. AB - Equations of state for completely unfolded proteins have been generated from group additivity algorithms and the revised Helgeson-Kirkham-Flowers (HKF) equations of state to compute the standard molal thermodynamic properties of these molecules at elevated temperatures and pressures. The requisite equations of state parameters were computed from those of groups retrieved by regression of experimental calorimetric and densimetric data reported in the literature. This approach permits calculation of the standard molal thermodynamic properties as a function of temperature and pressure for any completely unfolded protein for which the amino acid sequence is known. Calculations of this kind have been carried out for 11 thermophilic proteins. The thermodynamic properties reported below can be combined with those for protein unfolding to compute the corresponding properties of completely folded (i.e. native) proteins. PMID- 10796028 TI - Thermal activation of DNA frayed wire formation. AB - Dynamic light scattering has been used to study the formation of stable multistranded DNA complexes called frayed wires. DNA frayed wires arise from the indefinite self-association of oligonucleotides with long terminal tracks of guanines, e.g. d(A15G15). The complexes are stabilized via guanine-guanine interactions resulting in the formation of a guanine stem. Non-guanine portions of the oligonucleotide are disposed away from the stem and form single stranded arms. The indefinite nature of the self-association of these oligonucleotides leads to a distribution of aggregate molecular weights. The distribution arises from aggregated species that differ from one another by the number of self associated oligonucleotides. In light-scattering experiments, the autocorrelation functions collected for frayed wires are bimodal. The slow mode, often observed for samples containing DNA and other polyelectrolytes, has been ascribed to the formation of large unspecific aggregates due to electrostatic or other long-range interactions. We attribute the fast mode to the translational diffusion of the polydisperse population in the frayed wire sample. We use the mean of the fast mode to characterize the growth of the frayed wires. Consistent with the gel electrophoresis studies, the aggregation of frayed wires is promoted by the presence of magnesium ions and incubation at high temperature. The rate of aggregate formation increases with temperature, indicating the positive activation energy for the reaction. We propose an energy diagram for the formation/disruption of frayed wires revealing the catalytic-like role of the complementary strand in the denaturation of high molecular weight complexes. PMID- 10796029 TI - The resonant recognition model (RRM) predicts amino acid residues in highly conserved regions of the hormone prolactin (PRL). AB - The resonant recognition model (RRM) is a model which treats the protein sequence as a discrete signal. It has been shown previously that certain periodicities (frequencies) in this signal characterise protein biological function. The RRM was employed to determine the characteristic frequencies of the hormone prolactin (PRL), and to identify amino acids ('hot spots') mostly contributing to these frequencies and thus proposed to mostly contribute to the biological function. The predicted 'hot spot' amino acids, Phe-19, Ser-26, Ser-33, Phe-37, Phe-40, Gly 47, Gly-49, Phe-50, Ser-61, Gly-129, Arg-176, Arg-177, Cys-191 and Arg-192 are found in the highly conserved amino-terminal and C-terminus regions of PRL. Our predictions agree with previous experimentally tested residues by site-direct mutagenesis and photoaffinity labelling. PMID- 10796030 TI - Detection of the metastable rippled gel phase in hydrated phosphatidylcholine by fluorescence spectroscopy. AB - Steady-state and time-resolved emission spectroscopy of 1-anilinonaphthalene-8 sulfonic acid (ANS) have been used for characterization of the metastable rippled gel phase, Pbeta'(mst), formed in fully-hydrated dipalmitoylphosphatidylcholine (DPPC) upon cooling from the liquid crystalline phase Lalpha [Tenchov et al., Biophys. J. 56 (1989) 757]. The Pbeta'(mst) phase of DPPC clearly differs from the stable Pbeta' phase by increased (approximately 27%) ANS emission intensity, by enhanced (approximately 23%) average radiative rate constant, and by reduced (approximately 18%) non-radiative quenching rate constant. The fluorescence intensity peak at the Pbeta'-->Lalpha transition temperature is replaced by a large, reversible stepwise intensity drop at the Pbeta'(mst)-->Lalpha transition. No such effects have been found for dimiristoylphosphatidylcholine (DMPC) dispersions confirming previous results that DMPC does not form a Pbeta'(mst) phase. Since ANS is known to predominantly reside in the interfacial region, the observed effects indicate differences between the stable and metastable rippled phases in the organization and dynamics of their lipid/water interfaces. The data demonstrate that the metastable rippled phase manifests its appearance also through interactions with small molecules (ANS size approximately 8 A). PMID- 10796031 TI - Pyruvate kinase from the thermophilic eubacterium Bacillus acidocaldarius as probe to monitor the sodium concentrations in the blood. AB - We describe the isolation and characterization of a pyruvate kinase from the thermophilic eubacterium Bacillus acidocaldarius. This protein appears to be a tetramer composed of four 55-kDa subunits. The intrinsic tryptophan fluorescence of this protein is quenched by approximately 20% upon binding sodium, which occurs with a dissociation constant near 15 mM. Importantly, the intrinsic fluorescence of this pyruvate kinase does not appear to be affected by potassium, magnesium, and calcium at the concentrations found in whole blood. It appears that this pyruvate kinase can provide the basis for a selective protein sensor for sodium with minimal interference from other cations. PMID- 10796032 TI - Interactions of proteins in aqueous electrolyte solutions from fluorescence anisotropy and circular-dichroism measurements. AB - Understanding aqueous protein-protein interactions is crucial for the development of a molecular-thermodynamic model for salt-induced protein precipitation. In addition, protein interactions are important in many disease states, including cataract formation and alpha-amyloid diseases. Fluorescence anisotropy provides a means to measure intermolecular interactions. In this work, monomer-dimer equilibrium of the peptide T4 LYS(11-36) was studied by fluorescence anisotropy over the pH range 4-7 and the NaCl concentration range 0.0-1.0 M, in a 25 mM sodium phosphate buffer. This 26 amino-acid peptide is derived from the beta sheet region of the T4 lysozyme molecule and has the potential to form amyloid fibrils. The association constant for dimerization increases with rising pH and ionic strength. The potential of mean force for peptide-peptide interactions was calculated from these association constants. Circular-dichroism measurements show that the peptide becomes more structured as the pH rises, possibly contributing to increased association. PMID- 10796033 TI - The direction of glaucoma surgery: why improvements are occurring so slowly. PMID- 10796034 TI - Trabeculotomy in congenital glaucoma. AB - BACKGROUND: Congenital glaucoma is a potentially blinding disease that requires surgical therapy. This paper describes the outcome of trabeculotomy in primary congenital glaucoma. METHODS: Thirty-nine eyes of 22 children with congenital glaucoma who underwent trabeculotomy with or without a simultaneous trabeculectomy between 1992 and 1997 were retrospectively analyzed. RESULTS: Mean follow-up was 24.7+/-17.9 months. A mean of 1.3 operations per eye were performed. The mean IOP at the end of follow-up (n=39) was 17.7+/-6.0 mmHg; in 8 eyes (20.5%) the IOP was >21 mmHg, in 31 eyes (79.5%) it was < or =21 mmHg. The mean difference between pretreatment IOP and IOP at the end of follow-up (n=39) was -10.5+/-9.4 mmHg (-37.2%). Success rates were calculated: IOP was < or =21 mmHg in 36/39 eyes (92.3%) after 1/2 year of follow-up, in 25/27 eyes (92.6%) after 1 year, in 15/18 eyes (83.3%) after 2 years, in 8/12 (66.7%) eyes after 3 years, in 4/8 eyes (50%) after 4 years and in 4/4 (100%) eyes after 5 years of follow-up. Complications included hypotony (three eyes), subchoroidal bleeding (one eye ), detachment of Descemet's membrane (one eye) and macular pucker (one eye in which later mitomycin C was used). Visual acuity (VA) was tested with various methods in 35 eyes. VA was within the normal nomogram range in 12 eyes and below the normal range in 23 eyes at the end of follow-up. Axial length measurements showed normalization according to the age nomogram in 22 of 35 eyes. CONCLUSION: This study shows that trabeculotomy is an effective surgical procedure in congenital glaucoma with satisfactory success rates up to 5 years of follow-up. PMID- 10796035 TI - Photodynamic therapy of choroidal hemangioma: two case reports. AB - BACKGROUND: Photocoagulation, cryotherapy and radiotherapy have been used to treat angiomatous lesions. Depending on the location of the angioma, these treatments can cause additional, significant functional damage. Photodynamic therapy (PDT) however, allows a selective occlusion of vascular lesions without damaging adjacent retinal structures. METHODS: Two patients with isolated choroidal hemangiomas involving the posterior pole were treated with PDT. Treatments were performed using a diode laser at 692 nm, a light dose of 100 J/cm2 and 6 mg/m2 body surface area verteporfin (BPD-MA). PDT was applied in two courses in one eye and in four in the other eye at 1-4 months intervals. Patients were followed up for 9-12 months with visual acuity measurements and dilated ophthalmoscopy. Ultrasound, indocyanine green angiographic and fluorescein angiographic images were evaluated at each visit. RESULTS: Tumor heights were 3.3 and 4.6 mm on pretreatment ultrasound. After therapy, patients with isolated choroidal hemangioma showed total regression of the lesion and improved visual acuity due to resorption of retinal edema. Serous retinal detachment and cystoid macular edema resolved. Ultrasound demonstrated a progressive decrease in tumor height after each PDT application, with complete disappearance of the lesion. Retinal vessels were not affected by the treatment, and retinal function recovered in areas with previous tumor involvement. CONCLUSION: PDT allows selective treatment of large intraocular angiomatous lesions. Without optimized parameters, complete regression of choroidal hemangiomas, resolution of secondary complications and improvement of visual acuity were documented. PMID- 10796036 TI - Intraocular antibody production in intraocular inflammation. AB - BACKGROUND: The production of intraocular antibodies is considered a specific marker for active infectious uveitis. The aim of our study was to evaluate the diagnostic value of aqueous humor analysis in consecutive patients referred to a tertiary clinical center. METHODS: We analyzed 91 paired aqueous humor/serum samples from 89 patients with intraocular inflammation. In 71 patients aqueous humor analysis was used as a positive or negative confirmation of the suspected cause, whereas in 18 patients the clinical diagnosis was completely uncertain. A modified micro-ELISA technique was used to detect intraocular IgG production against Toxoplasma gondii, varicella zoster virus, herpes simplex virus and cytomegalovirus. Statistical analysis was performed using the "Cohen's kappa" test. RESULTS: Specific intra-ocular antibody production could be detected in 12 (66.7%) of 18 patients with uncertain diagnosis. Subsequently initiated therapy led to clinical improvement in 10 patients, whereas 2 patients remained unchanged. In 2 (2.8%) of 71 patients aqueous humor analysis led to revision of the initially suspected etiology and to a change of therapy. Statistical analysis showed a significant accordance of diagnosis and aqueous humor analysis (P<0.01). CONCLUSION: In patients with infectious uveitis, analysis of intraocular synthesis of specific antibodies is a valuable tool to establish the etiology rapidly and allows initiation of targeted antimicrobial treatment. PMID- 10796037 TI - Analgesic effects of sub-Tenon's versus retrobulbar anesthesia in planned extracapsular cataract extraction. AB - BACKGROUND: A prospective study was conducted to compare the analgesic effects of sub-Tenon's and retrobulbar anesthesia by assessing patients' response to the visceral stimulus. METHODS: Seven hundred and twenty eyes of 720 patients underwent extracapsular cataract extraction and posterior chamber intraocular lens implantation. They received retrobulbar anesthesia with 3 ml lidocaine (225 eyes), retrobulbar anesthesia with 5 ml lidocaine (216 eyes), or sub-Tenon's anesthesia with 3 ml lidocaine (279 eyes). Pain scores were recorded when an acetylcholine chloride solution was injected into the anterior chamber to attain miosis after lens implantation. RESULTS: Pain scores were significantly different among the three anesthesia groups (P<0.0001, Kruskal-Wallis test). The multiple comparison revealed that analgesic effects were highest with sub-Tenon's anesthesia, followed by 5-ml retrobulbar and 3-ml retrobulbar anesthesia. CONCLUSION: Sub-Tenon's anesthesia is an effective and reliable anesthetic method in cataract surgery. PMID- 10796038 TI - The effect of mitomycin C trabeculectomy on the progression of visual field defect in normal-tension glaucoma. AB - BACKGROUND: We investigated in a prospective fashion the visual prognosis and complications in normal-tension glaucoma following unilateral trabeculectomy with adjunctive mitomycin C. METHODS: Trabeculectomy with adjunctive mitomycin C was carried out unilaterally in 21 cases of normal-tension glaucoma. Intraocular pressure (IOP), visual prognosis, and complications were compared between the operated eyes and the non-operated fellow eyes. The follow-up period ranged from 2 to 7 years. RESULTS: The IOP dropped significantly from 14.8+/-1.8 mmHg (mean +/- SD) to 9.6+/-3.9 mmHg in the operated eyes (P=0.0002, Wilcoxon signed-rank test), but did not drop in the non-operated eyes. The mean deviation (MD) was 12.69+/-6.41 dB preoperatively and -14.70+/-5.49 dB at the last clinic visit in the operated eyes, whereas in non-operated eyes it was -7.85+/-5.65 dB and 11.15+/-5.62 dB, respectively. The MD deteriorated significantly in both operated and non-operated eyes (operated eyes P=0.0239, non-operated eyes: P=0.0002; Wilcoxon signed-rank test). The MD slope was -0.37+/-0.60 dB/year and -0.71+/ 0.89 dB/year for the operated and non-operated eyes, respectively (P=0.5243, Mann Whitney U-test). Visual field deterioration was more frequently observed in the non-operated eyes by a pointwise definition of the progression (P<0.05, McNemar test). Visual acuity deteriorated in 6 of the operated eyes and in 5 of the non operated eyes. Cataract developed in 6 (29%) of the 21 operated eyes, while among the non-operated eyes 4 (19%) developed cataract. CONCLUSION: Mitomycin C trabeculectomy is effective in delaying progression of visual field defect in normal-tension glaucoma, but complications may arise and cause some visual disturbance. PMID- 10796039 TI - Elevated levels of calcitonin gene-related peptide in aqueous humor of patients with proliferative vitreoretinopathy. AB - PURPOSE: To detect the levels of the sensory peptide calcitonin gene-related peptide in aqueous humor of patients with proliferative vitreoretinopathy and to compare them with those of uninflamed eyes (cataract and uncomplicated rhegmatogenous retinal detachment). MATERIALS AND METHODS: Using a highly specific and sensitive radioimmunoassay the concentration of calcitonin gene related peptide was detected in fresh samples of aqueous humor obtained via paracentesis. Furthermore, calcitonin gene-related peptide-like immunoreactivities were characterized by high-pressure liquid chromatography. RESULTS: The mean level of calcitonin gene-related peptide was 6.11 fmol/ml in cataract controls and 14.77 fmol/ml in uncomplicated rhegmatogenous retinal detachment. In the cataract group, 9 of 18 cases were below the detection limit and in the retinal detachment group, 5 of 16. In proliferative vitreoretinopathy, the peptide averaged 76.92 fmol/ml and none of the samples was below the detection limit. High-pressure liquid chromatography revealed one major peak corresponding to synthetic calcitonin gene-related peptide. CONCLUSION: In recent studies, we found elevated levels of the sensory peptide substance P in aqueous humor of patients with proliferative vitreoretinopathy. This fact and the present result, the elevation of calcitonin gene-related peptide in aqueous humor of eyes with proliferative vitreoretinopathy, clearly point to an involvement of sensory peptides in the pathobiology of the disease. The source of the elevation is not clear, but we hypothesize that it originates from a neurogenic mechanism, i.e. an acceleration of the peptides by their enhanced release from the iris/ciliary body complex subsequent to sensitization of sensory neurons, thus representing a very interesting epiphenomenon of proliferative vitreoretinopathy. Our results constitute novel aspects in the pathophysiological concept of proliferative vitreoretinopathy and extend the knowledge about the pathobiology of the disease process. PMID- 10796041 TI - Transpupillar laser phototherapy for retinal and choroidal tumors: a rational approach. AB - BACKGROUND: The physical laws are considered that need to be taken into account for optimal photothermal treatment of solid and vascular tumors, as well as other vascular anomalies of various etiology of the retina and choroid. Optimal irradiation therapy should take into account the distribution of both radiant and thermal energy in tumors, such as retinoblastomas, malignant melanomas and vascular malformations. Strict confinement of the extent of photothermal damage is critical since such pathological entities are frequently located close to the macula or optic nerve head. METHODS: A formal treatment of the optical quantities related to these requirements is presented. In this analysis we emphasize the following topics: Arrhenius' law, the kinetics of protein denaturation, the electromagnetic radiation field, wavelength, laser pulse duration (exposure time), optical properties of tissue, photocoagulation and thermotherapy. RESULTS: Generally, the conditions are best fulfilled when using radiation in the near infrared range of the electromagnetic spectrum, such as that emitted from the diode (810 nm) and Nd:YAG (1064 nm) laser, because of the good optical penetration properties of this radiation in tissue. The xenon arc lamp was a very effective and particularly appropriate energy source for such purposes, and its withdrawal from the world market may have been untimely. Short wavelength sources of radiation, such as the argon ion (488, 514 nm) or the frequency-doubled Nd:YAG (532 nm) laser, are unsuitable for the irradiation of large vascular structures, as they have poor penetration depths. However, for vascular formations with a short path length (1 mm or less), short wavelength sources appear to be the most appropriate choice. Optical coupling of radiant energy to the eye by means of indirect ophthalmoscopic systems or contact lenses to the eye is crucial. Strong positive lenses may lead to severe constriction of the laser beam which leads to high irradiance within the anterior segment, increasing the chances of it being damaged; with negative contact lenses, such as the -64 D Goldmann type, this danger is reduced. CONCLUSIONS: Photothermotherapy is not without risk unless the temperature field can be well adapted to the tumorous structure, as temperature elevations outside a small therapeutic range that affect vital structures are considered to be a risk factor. PMID- 10796040 TI - Involvement of NMDA-receptor in kainate-induced neurotoxicity in cultured fetal retinal neurons. AB - BACKGROUND: Both in vivo and in vitro studies suggest that excess stimulation of non-NMDA receptors can result in massive neuronal death in the retina. In particular, murine amacrine neurons have been known to show marked susceptibility to the toxic effects of kainate. PURPOSE: This study was designed to examine and characterize the role of N-methyl-D-aspartate (NMDA) receptor vs non-NMDA receptor in glutamate-induced neurotoxicity in the retina. METHODS: Primary cultures obtained from fetal rat retina (gestation day 16-19) were used for the experiment. The neurotoxicity was assessed quantitatively using the trypan blue exclusion method. Electrophysiological studies using patch-clamp techniques were performed to record whole-cell currents evoked by these excitatory amino acids. RESULTS: Removal of extracellular Ca2+ from the medium or application of MK-801 reduced the extent of cell death induced by the brief exposure to glutamate, NMDA, and kainate. By contrast, cell death induced by a 60-min exposure to kainate was not affected by MK-801. The electrophysiological study demonstrated that MK-801 abolished the whole-cell currents evoked by NMDA but had no effect on those induced by kainate or AMPA. CONCLUSION: These findings demonstrate that brief exposure to kainate induces cell death by way of activating NMDA receptors in cultured fetal retinal neurons and that NMDA receptors are the predominant route of fetal retinal neurotoxicity induced by brief glutamate exposure. PMID- 10796042 TI - Inhibition of diclofenac formulated in hyaluronan on angiogenesis in vitro and its intraocular tolerance in the rabbit eye. AB - PURPOSE: To investigate the effect of diclofenac, a potent nonsteroidal anti inflammatory drug, formulated in hyaluronan (diclofenac/HA) on angiogenesis in vitro and its intraocular toxicity in vivo. METHODS: The effect of diclofenac/HA on angiogenesis was determined by choriocapillary endothelial cells on Matrigel stimulated by vascular endothelial growth factor (VEGF). The tube areas were quantified by image digital analysis. For toxicity study, diclofenac/HA was injected intravitreally with a dose range from 100 to 1080 microg in 26 rabbits following gas compression vitrectomy. Potential toxicity was assessed by indirect ophthalmoscopy and by histological studies (light and electron microscopy). Retinal function was monitored by electroretinography (ERG) in six rabbits that received 400 microg of diclofenac/HA. RESULTS: Diclofenac/HA, 180, 90 microg/ml, inhibited tube formation to 24% and 55% of the standard group (Media Ham's F12 plus 5% fetal calf serum and 50 ng/ml VEGF) respectively (P<0.01). Intravitreal injection of 540 microg or higher doses of diclofenac/HA resulted in ocular toxicity in the rabbit, demonstrated as cataract, vitreous haze and retinal damage observed by indirect ophthalmoscopy and light- and electron-microscopic examinations. No toxicity was observed in the eyes that received 400 microg or less diclofenac/HA, which was further supported by the normal ERG examined at 4 and 25 days post injection. CONCLUSIONS: Diclofenac/HA inhibits tube formation in vitro and is non-toxic to the rabbit retina at concentrations that are inhibitory to tube formation. Our results suggest diclofenac/HA may be an effective candidate to inhibit ocular neovascularisation related to granulomatous reaction in the eye. PMID- 10796043 TI - Immunolocalization of TGF-beta1, -beta2, and -beta3, and TGF-beta receptors in human lens capsules with lens implants. AB - PURPOSE: To establish the alteration in expression pattern of transforming growth factor (TGF)-betas and their receptors during repair of lens capsules after cataract surgery, we immunohistochemically located TGF-beta isoforms and their receptors in human lens capsules before and after cataract surgery. METHODS: Ten post-cataract surgery capsular specimens were obtained during vitrectomy. Three sections of the anterior capsules were obtained during cataract surgery. A whole lens capsular bag immediately after lens extraction was obtained during vitrectomy. Cryosections of these specimens were processed for immunohistochemical analysis for TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta receptor type I (TbetaR-I), type II (TbetaR-II) and type III (TbetaR-III), and were observed under light micros-copy. RESULTS: Lens epithelial cells (LECs) lining the inner surface of the anterior capsules exhibited immunoreactivity for TGF-beta2 and TbetaR-II. Immunoreactivity for TGF-beta1, -beta3, TbetaR-I and TbetaR-III was negative. In the whole capsular bag specimen, equatorial LECs were positive for TGF-beta1 and -beta2, but not for -beta3. In post-cataract surgery specimens, antibodies for each TGF-beta isoform labelled either the LECs or ECM accumulated on the capsules. Post-surgical LECs expressed TbetaR-I and TbetaR-II, and had also TbetaR-III in seven of the nine specimens examined. CONCLUSION: Expression pattern of TGF-beta s in quiescent LECs showed regional heterogeneity. Anterior LECs exhibited TGF-beta2 immunoreactivity, while equatorial LECs were positive for TGF-beta1 and -beta2. Quiescent LECs expressed TbetaR-II. LECs proliferating around IOLs expressed proteins of each TGF-beta isoform and each TbetaR. TGF-beta s were also localized in the ECM on capsules undergoing repair. TGF-beta3, TbetaR-I and TbetaR-III are up-regulated in LECs after cataract surgery. PMID- 10796044 TI - Helicobacter pylori infection and carcinogenesis of the stomach. AB - INTRODUCTION: Human stomach carcinogenesis occurs after a multi-step process of genetic and epigenetic alterations in oncogenes, tumor-suppressor genes, cell adhesion molecules, telomere and telomerase activity as well as genetic instability at several microsatellite loci. RESULTS AND DISCUSSION: These sequential alterations found in gastric cancer differ between the two histological types, indicating that different genetic pathways exist for well differentiated or intestinal-type and poorly differentiated or diffuse-type gastric cancers, even though both types of gastric cancer may arise from epithelial "stem cells", which express human telomerase reverse transcriptase (hTERT) protein and telomerase activity. Infection with Helicobacter pylori, which evidently causes the release of reactive oxygen species (ROMs) and reactive nitrogen species (NO), may be a strong trigger for "stem cell" hyperplasia in intestinal metaplasia, followed by telomere reduction and increased telomerase activity as well as hTERT overexpression. They may precede DNA replication error, DNA hypermethylation, CD44 abnormal transcript, and p53 mutations, all of which occur in at least 30% of intestinal metaplasias as early events of multi-step pathogenesis of well-differentiated type gastric cancer. Here, we propose a new concept for gastric preneoplasic lesion, "metaplastic dysplasia", based on our molecular observations. PMID- 10796045 TI - Helicobacter pylori infections--are these diseases relevant for surgical treatment? AB - Helicobacter pylori infection and its strong association with gastroduodenal diseases has revolutionized our views and treatment options in these diseases. Surgery has always been considered a mainstay in their treatment. The development of potent eradication regimes has changed indications for operative treatment. With endoscopic procedures being increasingly performed by the surgeon himself, it has become necessary that he educate himself in the diagnosis and treatment of H. pylori infection in order to be able to cope with these diseases in every day surgical practice. Treatment of H. pylori infection may not only be seen as an alternative to operative therapy, but also as an adjunct following surgical treatment of gastroduodenal disease. PMID- 10796046 TI - Surgical management of peptic ulcer disease today--indication, technique and outcome. AB - AIMS: The current surgical management of peptic ulcer disease and its outcome have been reviewed. RESULTS: Today, surgery for peptic ulcer disease is largely restricted to the treatment of complications. In peptic ulcer perforation, a conservative treatment trial can be given in selected cases. If laparotomy is necessary, simple closure is sufficient in the large majority of cases, and definitive ulcer surgery to reduce gastric acid secretion is no longer justified in these patients. Laparoscopic surgery for perforated peptic ulcer has failed to prove to be a significant advantage over open surgery. In bleeding peptic ulcers, definitive hemostasis can be achieved by endoscopic treatment in more than 90% of cases. In 1-2% of cases, immediate emergency surgery is necessary. Some ulcers have a high risk of re-bleeding, and early elective surgery might be advisable. Surgical bleeding control can be achieved by direct suture and extraluminal ligation of the gastroduodenal artery or by gastric resection. Benign gastric outlet obstruction can be controlled by endoscopic balloon dilatation in 70% of cases, but gastrojejunostomy or gastric resection are necessary in about 30% of cases. CONCLUSIONS: Elective surgery for peptic ulcer disease has been largely abandoned, and bleeding or obstructing ulcers can be managed safely by endoscopic treatment in most cases. However, surgeons will continue to encounter patients with peptic ulcer disease for emergency surgery. Currently, laparoscopic surgery has no proven advantage in peptic ulcer surgery. PMID- 10796047 TI - Helicobacter pylori in the upper gastrointestinal tract: medical or surgical treatment of gastric lymphoma? AB - The treatment of primary gastric lymphoma is controversial. The role of surgery has come to be questioned with increasing knowledge about the pathogenesis of gastric lymphoma and with new therapeutic approaches such as eradication of Helicobacter pylori. We review published clinical trials of primary gastric lymphoma, including preliminary results of our own prospective multicenter trial. The results of 7 trials of H. pylori eradication and 12 prospective therapeutic trials trial are discussed. On basis of these data it is concluded that surgery with intention of R0 resection is the treatment of choice in stages EI2 and EII1 of low-grade lymphoma. In high-grade lymphomas it is still unclear whether surgery or its primary combination with radio- or chemotherapy should be preferred. The eradication of H. pylori is a promising therapeutic approach for localized low-grade mucosa-associated lymphoid tissue lymphoma. A randomized trial is needed to clarify whether medical or surgical management of localized gastric lymphoma or a combination of two is the best treatment modality. PMID- 10796048 TI - Unilateral open and minimally invasive procedures for primary hyperparathyroidism: a review of selective approaches. AB - The currently established procedure for surgical treatment of primary hyperparathyroidism is bilateral exploration and visualization of all four glands to identify an adenoma and exclude multiglandular disease. With the development of improved preoperative localization imaging of the parathyroids using high resolution ultrasonography and sestamibi scintigraphy, on the one hand, and perioperative control of surgical success with a rapid parathyroid hormone assay on the other, unilateral and minimally invasive techniques have become feasible. For patients with unequivocal localization in preoperative sestamibi scintigraphy and high-resolution ultrasonography of the parathyroid adenoma in probable single gland disease, the unilateral and minimally invasive parathyroidectomy present a therapeutic option. Perioperative rapid parathyroid hormone assays, although costly, offer immediate supervision of adenoma extirpation and differentiation of single- and multiglandular disease. These methods demonstrate advantages with favorable cosmetic results and lower reported rate of postoperative hypoparathyroidism. These methods are already being practiced in some places under local anesthesia and in an ambulatory setting. This contribution provides an introduction and overview of the currently practiced unilateral and minimally invasive techniques of parathyroidectomy for primary hyperparathyroidism, discussing indications, advantages and disadvantages, and technical differences in the practiced methods. PMID- 10796049 TI - Laparoscopic management of traumatic ruptures of the diaphragm. AB - INTRODUCTION: As a result of the relatively high frequency of high-speed accidents, diaphragmatic rupture is a diagnosis that is increasingly being established. Not all of these, usually multi-traumatized, patients are diagnosed as having diaphragmatic rupture immediately following the traumatic event- rather, an appreciable number of these injuries are not detected until some time later--often after a considerable delay. Most of the cases involve rupture of the left diaphragm, with most defects occurring in the region of the central tendon. METHODS: During the course of the second half of the year 1998, we operated on three patients with left diaphragmatic rupture. Two of these patients were treated immediately following traumatization, while the third case was a 10-year old rupture originally misdiagnosed as a para-esophageal hernia. In all three cases, we were able to reduce the hernia and close the diaphragmatic defect laparoscopically. In the case of the two patients with a fresh rupture, the post operative course was unremarkable, while in the patient with the missed rupture, a serous pleural effusion requiring drainage occurred on the left side. CONCLUSION: Overall, it would appear that in the case of an acute traumatic diaphragmatic rupture in particular, laparoscopic management, with its low level of traumatization and excellent access, offers a favorable alternative to conventional surgery. A point to be considered, however, is the fact that probably not every hospital will have the facilities for laparoscopic management available on a 24-h basis. In the case of longstanding ruptures, reduction of herniated bowel and treatment of the rupture will make considerable demands on the surgeon. PMID- 10796050 TI - Hemosuccus pancreaticus complicating chronic pancreatitis: an obscure cause of upper gastrointestinal bleeding. AB - BACKGROUND: Hemosuccus pancreaticus, a rare form of upper gastrointestinal bleeding, may complicate chronic pancreatitis and pose a significant diagnostic and therapeutic dilemma. AIM: To present our experience with this potentially life-threatening complication of chronic pancreatitis. METHODS: We reviewed our experience with management (both operative as well as angiographic embolization) of patients with hemosuccus pancreaticus complicating histologically documented chronic pancreatitis between 1976 and 1997. Diagnosis of hemosuccus pancreaticus was based on clinical presentation, preoperative endoscopic and radiographic imaging, operative findings, and pathologic evaluation. RESULTS: During the period, we managed eight patients with hemosuccus pancreaticus (1.5% of all patients with chronic pancreatitis treated surgically). Gastrointestinal bleeding presented as hematemesis in three and hematochezia in three, but all had recent melena and were anemic; three of these patients were hemodynamically unstable. Abdominal pain was present in six. When performed, angiography (n=6) was diagnostic of a pseudoaneurysm; computed tomography (n=7) showed a pseudoaneurysm in two and a pseudocyst in five. Endoscopy (n=8) revealed blood issuing from the ampullary papilla in two patients. Operative management (n=6) involved distal pancreatectomy, pancreatoduodenectomy, or total pancreatectomy in two patients each. Angiographic embolization was successful in one patient, but the other died from uncontrollable hemorrhage. CONCLUSIONS: Hemosuccus pancreaticus is rare, but should be considered in patients with chronic pancreatitis and gastrointestinal bleeding. In the absence of pancreatitis-related indications for surgery, angiographic embolization can be definitive treatment. If there are pancreatitis related indications for operation, angiographic embolization may allow an elective operative procedure based on structural changes of the pancreas. If embolization fails, pancreatic resection is usually required, often on an emergent basis. PMID- 10796051 TI - Impact of 18F-FDG-positron emission tomography for decision making in colorectal cancer recurrences. AB - Diagnostic imaging for suspected tumour recurrence of primary colorectal cancer frequently lacks specificity and sensitivity. The impact of whole body 18F-FDG positron-emission tomography (PET) on detection of local recurrences and hepatic or pulmonary metastases was evaluated in a prospective study. Results were compared with computed tomography (CT), ultrasonography, magnetic resonance imaging and conventional chest X-ray. The study included 71 patients (77 investigations) with suspected local recurrence, hepatic metastases or unexplained raised level of the tumour marker carcinoembryonic antigen (CEA). The results demonstrate that 18F-FDG-PET was clearly superior to CT with regard to detection of hepatic metastases. Sensitivity was 1.0 and specificity 0.98 compared with 0.87 and 0.91 for CT. In four cases, 18F-FDG-PET clarified otherwise unclear local recurrences. In five patients, 18F-FDG-PET showed pulmonary metastases that had previously been unknown. In a total of 16 patients (20.8%), 18F-FDG-PET provided additional information leading to a change of the treatment strategy. 18F-FDG-PET clearly has the ability to detect colorectal tumour recurrence and its metastases in a whole body format. Therefore, it may be applied in the follow-up of patients with primary colorectal cancer. Despite the costs, it is certainly recommended for patients with an otherwise unclear increase of CEA level or with unproven local recurrence. PMID- 10796052 TI - DNA polymerases and Ki-67 nuclear antigen are induced in correlation with the resected mass of rat liver up to 90%. AB - BACKGROUND AND AIMS: We studied the regeneration potential by measuring induction of DNA polymerases in the remnant rat liver after a partial hepatectomy (PHx) that is maximal but compatible with survival. METHODS: The regenerating rat liver was obtained after the 90% PHx. The induction of activities of DNA polymerase alpha, delta, and epsilon were measured after partial purification. The Ki-67 nuclear antigen was also detected histochemically. These parameters were compared with those after both 30% and 70% PHx. RESULTS: The 90% hepatectomy resulted in the strong inductions of DNA polymerase alpha, delta, and epsilon, at 48 h after operation, in association with increases in wet weight and total DNA in the remnant liver. The enzyme induction was much higher after 90% PHx than after 30% and 70% hepatectomy, in correlation with the resection volume. At 48 h after 90% hepatectomy, the Ki-67 positive cells increased up to 47.2% of hepatocytes in the remnant liver. CONCLUSION: The higher induction of replication enzymes by 90% hepatectomy reflects more cells entering mitogenic cell cycle, which supports the fast regeneration of the remnant liver. The number of proliferating hepatocytes is stringently controlled by an unknown mechanism sensing the mass of resected liver parenchyma. PMID- 10796053 TI - Evidence-based surgery: diverticulitis--a surgical disease? AB - Sigmoid diverticulitis is an increasingly common disorder. While there is no gender difference, the incidence increases with age. Many reports have been published on the topic, but there is no consensus on certain aspects of treatment. We conducted a literature search covering the past 30 years and report our own data. Two major areas of controversy exist. One concerns indications for elective surgery for symptomatic diverticulitis. The consensus is that there is no indication for prophylactic surgery. The first attack should be treated conservatively; elective surgery is considered following a second attack, but in immunocompromised patients earlier. The second controversy concerns surgical strategy in peritonitis from perforation. Three-stage operations have generally been abandoned. The question is whether to perform a sigmoid resection with primary anastomosis. One end of the spectrum is recent perforation which can be treated safely by resection and anastomosis. The other end is advanced feculent peritonitis in high-risk patients. In this situation a Hartmann procedure is recommended. Although data from prospective randomized studies are lacking, there seem to be indicators in the individual situation that allow a rational selection of the appropriate procedure. Diverticulitis can thus be treated surgically for a broad range of its forms of presentation. PMID- 10796054 TI - Inverse agonism at heptahelical receptors: concept, experimental approach and therapeutic potential. AB - Inverse agonists (negative antagonists) are ligands that stabilize the inactive conformation (R) of receptors according to the two-state receptor model. The active conformation (R*) of heptahelical receptors, i.e. G protein-coupled receptors, has high affinity for G proteins. According to ternary complex models of receptor activation, the R*G complex is in equilibrium with R + G, with spontaneous activity in the absence of agonist. Inverse agonists, having a higher affinity for R, shift R*G towards R + G, decreasing the spontaneous activity of receptors. Agonists have the opposite effect, with a higher affinity for R*. Neutral antagonists have the same affinity for R and R* and compete for both agonists and inverse agonists. Inverse agonists have been recently proposed for a variety of heptahelical receptors. Methods to detect inverse agonists among antagonists are based on the determination of ligand affinity at R and R* with binding experiments, and on the modulation of G protein activity (GTP binding and hydrolysis) or of effector activity. Receptor inverse agonists, but also G protein antagonists and GTPase inhibitors, decrease spontaneous G protein activity corresponding to R*G. Receptor agonists, G protein agonists and GTPase inhibitors increase effector basal activity, but receptor inverse agonists decrease it. The therapeutic potential of inverse agonists is proposed in human diseases ascribed to constitutively active mutant receptors and may be extended to diseases related to wild-type receptor over-expression leading to the increase of R*. Some of the therapeutic effects of presently used receptor antagonists may be related to their inverse agonist properties. Inverse agonists lead to receptor upregulation, offering new approaches to tolerance and dependence to drugs. PMID- 10796055 TI - Gastroprotective effects of pantoprazole against experimental mucosal damage. AB - The present study investigated the gastroprotective effects of the proton pump inhibitor pantoprazole on gastric mucosal damage induced by ethanol-HCl in rats. Omeprazole was used as reference drug. The morphometric analysis of gastric histological sections revealed that pantoprazole and omeprazole dose-dependently prevented the necrotic mucosal injury evoked by ethanol-HCl (ED50 = 14.1 and 21.6 micromol/kg, respectively). These effects were associated with a marked increment of Alcian blue recovery from gastric bound mucus (ED50 = 18.8 and 29.3 micromol/kg, respectively). In addition, both pantoprazole and omeprazole inhibited gastric acid secretion in pylorus-ligated rats (ED50 = 1.5 and 3.3 micromol/kg, respectively). Further experiments indicated that the protective effects of pantoprazole were not modified by L-365,260 (a gastrin receptor antagonist), suramin (a drug able to interfere with endogenous growth factors), N(G)-nitro-L-arginine (an inhibitor of nitric oxide synthase) or systemic ablation of capsaicin-sensitive sensory nerves, whereas they were partly blocked by indomethacin (an inhibitor of prostaglandin synthesis) and fully prevented by N-ethylmaleimide (a potent blocker of sulfhydryl compounds). The present data provide histomorphometric evidence that: 1) pantoprazole is endowed with gastroprotective properties and is more active than omeprazole in preventing the necrotic mucosal damage induced by ethanol-HCl; 2) according to the rank order of ED50 values, the protective effects of both drugs appear to depend mainly on the enhancement of the gastric mucosal barrier rather than on the inhibition of acid secretion; 3) an increased production of prostaglandins, as well as an increased availability of sulfhydryl radicals at the level of the gastric mucosa may account for the gastroprotective effects of pantoprazole. PMID- 10796056 TI - Angiotensin II receptor blockade unmasks a depressor response to endothelin antagonists in rats. AB - Endothelin (ET) antagonists do not decrease blood pressure in normal rats. Since angiotensin II (AII) and ET both induce smooth muscle cell contraction through the same transduction pathways we designed experiments to assess whether blockade of the renin angiotensin system would unmask a vasodilatory response to ET receptor antagonists in rats. For this purpose, we tested the effect on arterial blood pressure of the mixed ETA-ETB receptor antagonist bosentan or of the ETA antagonist BQ-123 in the absence or the presence of the AII receptor antagonist losartan. In control conditions bosentan did not affect arterial blood pressure. In contrast, in losartan-pretreated rats, bosentan induced a marked, dose dependent decrease in arterial pressure (% change after bosentan 10 mg/kg: control -3 +/- 3, losartan -32 +/- 6; cilazapril -28 +/- 3). Similarly, BQ-123 decreased blood pressure in losartan-pretreated but not in control rats. Bosentan also increased the hypotensive effect of losartan in conscious, normotensive rats. The hypotensive effect of the combination of bosentan and losartan was not associated with any changes in cardiac output or heart rate, and thus was entirely due to a decrease in total peripheral resistance. We conclude that blockade of angiotensin II, AT1 receptors unmasks a vasodilator response to ET antagonists. This suggests that endogenous ET plays an active role in the maintenance of arterial blood pressure in rats which can be unmasked by a concomitant inhibition of the renin angiotensin system. PMID- 10796057 TI - Frequency-independent blockade of cardiac Na+ channels by riluzole: comparison with established anticonvulsants and class I anti-arrhythmics. AB - The Na+ channel blocking activity and the antiarrhythmic effects of riluzole, and established anticonvulsants (lamotrigine and lifarizine) and class I antiarrhythmics (lidocaine, flecainide and disopyramide) were studied under in vitro and in vivo conditions. Guinea-pig cardiac Purkinje fibres were superfused with Tyrode solution and electrically driven for recording action potentials with intracellular microelectrodes. In these preparations paced at 1 Hz, all compounds tested produced concentration-dependent (0.3-100 microM) reductions in the maximum rate of depolarization of the action potential (Vmax). For riluzole, phenytoin and carbamazepine this effect was frequency-independent (0.5-6 Hz) but for lamotrigine, lifarizine, lidocaine, flecainide and disopyramide it was frequency-dependent. In anaesthetized rats, riluzole, in contrast to flecainide, did not delay the appearance of aconitine-induced arrhythmias. Riluzole (0.3-3.9 mg/kg, i.v.) also lacked notable cardiac electrophysiological effects in anaesthetized dogs. At an i.v. dose of 3.0 mg/kg riluzole failed to restore a normal sinus rhythm in conscious dogs with polymorphic arrhythmias produced by ligation of the left anterior descending coronary artery 24 h earlier. These results indicate that riluzole, phenytoin and carbamazepine, unlike lamotrigine, lifarizine and flecainide, block cardiac Na+ channels in a frequency-independent manner. This property may account for the lack of antiarrhythmic activity of riluzole, phenytoin and carbamazepine in animal models of arrhythmias that respond to class I antiarrhythmic drugs. It may also account for the clinical observation that riluzole does not seem to cause the unfavourable electrocardiographic changes characteristic of drugs that block cardiac Na+ channels in a frequency-dependent manner. PMID- 10796058 TI - Hypertension in transgenic (mREN2)27 rats is not associated with the presence of B1 receptors. AB - B1 receptors are inducible receptors expressed only in stressful conditions. The aim of this study was to determine if, in (mREN2)27 transgenic rats, hypertension is associated with the presence of B1 receptors in the cardiovascular system and if a heat stress inducible effect is preserved during hypertension. Age-matched (16 weeks old) heterozygous hypertensive transgenic (mREN2)27 rats (HT rats) and the normotensive control animals (homozygous Sprague-Dawley rats, NT rats) were used. The study was conducted in two parts: in the first part the responsiveness of B1 receptors was studied in rats submitted to heat stress (42 degrees C rectal temperature, 20 min) or sham anaesthesia 24 h before, by recording changes in isometric tension in aortic rings in response to [des-Arg9]-bradykinin, a B1 receptor agonist. In the second part, we studied whether B1 receptor mRNA was present in aorta, heart and kidneys, using a semi-quantitative RT-PCR technique. [des-Arg9]-Bradykinin induced a concentration-dependent relaxation of aortic rings only from animals submitted to prior heat stress. This response was significantly higher in aortic rings from heat stressed HT rats than from heat stressed NT ones. B1 receptor mRNA was undetectable in organs from rats not submitted to heat stress but they were present 5 h after heat stress in aorta, heart and kidneys from both NT and HT rats. In conclusion, arterial hypertension observed in (mREN2)27 rats is not associated with the presence of B1 receptors. However, after heat stress, we observed an increase in responsiveness from HT rat aortas compared to NT ones. PMID- 10796060 TI - Influence of low temperature on bronchodilatation induced by terbutaline administered by metered dose or dry powder inhalers in asthmatics. AB - Low temperatures may affect dose delivery efficacy and clinical effectiveness of medication aerosols. In this study we examine the effect of cold ambient temperature on the bronchodilatation produced by terbutaline delivered from a chlorofluorocarbon pressurized metered dose inhaler (pMDI) compared to a multi dose dry powder inhaler (DPI). Fourteen stable asthmatics were studied on two consecutive days. On day 1, after measuring FEV1 at room temperature (22 degrees C), each patient was randomized to receive 500 microg of terbutaline delivered from pMDI or DPI stored for 24 h at 22 degrees C with FEV1 recorded 20 min post dose; then, patients were placed in a chamber at -10 degrees C, and after obtaining FEV1, each patient received 500 microg of terbutaline delivered from pMDI or DPI (same formulation as previously administered) stored for 24 h at -10 degrees C, and FEV1 was obtained 20 min post-dose. On day 2, a similar protocol was followed but each patient received terbutaline as the alternative to the formulation administered on day 1. Pairwise comparisons of the FEV1 (% predicted) values obtained on day 1 and day 2 at 22 degrees C and -10 degrees C (pre-dose) showed no significant differences. Similar bronchodilatations were observed for terbutaline DPI administration at 22 degrees C and -10 degrees C (24.85 +/- 11.72 and 20.08 +/- 6.27% increase of FEV1; P > 0.05). By contrast, the bronchodilatation obtained for terbutaline pMDI at 22 degrees C (21.07 +/- 8.55% increase in FEV1) was not reproduced at -10 degrees C (0.72 +/- 2.84%; P < 0.05 from 22 degrees C). In five asthmatics a cumulative dose-response curve for terbutaline pMDI was obtained. This part of the study showed that a higher dose of terbutaline pMDI was necessary at -10 degrees C to obtain a bronchodilator response (10.04 +/- 6.75% increase of FEV1 after 2,000 microg) that remained lower than the bronchodilatation for 500 microg terbutaline pMDI at -10 degrees C. In conclusion, the clinical effectiveness of terbutaline delivered from chlorofluorocarbon pMDIs is compromised by cold storage while DPIs are not affected. PMID- 10796059 TI - Attenuation of the acute amnestic effect of ethanol by ifenprodil: comparison with ondansetron and dizocilpine. AB - The aim of the present study was to examine the influence of ifenprodil, a non competitive N-methyl-D-aspartate (NMDA) receptor antagonist which also blocks 5 HT3 receptors, on the amnestic effect of ethanol in a passive avoidance task in mice. The anti-amnestic action of ifenprodil was compared with the effects of the 5-HT3 receptor antagonist ondansetron and the non-competitive NMDA-receptor antagonist dizocilpine (MK-801). Ethanol, 2 g/kg and dizocilpine 0.1 mg/kg significantly impaired the passive avoidance response. In contrast, ifenprodil (0.1-10 mg/kg), ondansetron (0.03-0.3 mg/kg) and dizocilpine (0.01 and 0.03 mg/kg) did not alter passive avoidance by themselves. Dizocilpine did not diminish the amnestic action of ethanol when administered at doses of 0.03-0.1 mg/kg. However, the amnestic effect of ethanol was partially restored towards normal by ifenprodil 0.3 mg/kg and by ondansetron 0.03 mg/kg (alone or together with dizocilpine 0.01 mg/kg) but it was not affected by other doses of ifenprodil (0.1, 1 and 10 mg/kg) and ondansetron (0.1 and 0.3 mg/kg). In conclusion, ifenprodil at an appropriate dose reduced ethanol-induced amnesia in a step through passive avoidance task. The results are compatible with the assumption that the anti-amnestic action of ifenprodil may be (at least partially) due to an antagonism at 5-HT3 receptors. PMID- 10796061 TI - Pharmacokinetics of sustained and immediate release formulations of indapamide after single and repeated oral administration in healthy volunteers. AB - The pharmacokinetics of a 2.5 mg immediate release (IR) formulation of indapamide was compared to a 1.5 mg sustained release (SR) formulation of indapamide after single and repeated oral administration dose using double blind randomised cross over studies. In the first study, 12 subjects received a single dose of each treatment: IR fasted, SR fasted or with food. In the second study one tablet of either formulation was administered daily for one week at breakfast. In each study, blood samples were collected pre dose (Cmin) and up to 120 h after the last dose. Urine was collected over the dosing interval (24 h). Following a single oral administration the SR formulation had a lower dose-normalised Cmax compared to the IR formulation (17.6 +/- 6.3 vs. 39.3 +/- 11.0 ng x mL(-1), respectively), a much longer t(max) (12.3 +/- 0.4 vs. 0.8 +/- 0.3 h) and a greater t75 (15.3 +/- 6.1 vs. 1.8 +/- 1.4 h) but there were no differences in dose-normalised AUC (559 +/- 125 and 564 +/- 146 ng x h x mL(-1)) nor in t(1/2z) values (14.8 +/- 2.8 vs. 18.4 +/- 13.4 h). The SR formulation clearly demonstrated sustained release characteristics as compared to the IR formulation. Food co-administration had no effect on dose-normalised AUC for the SR formulation. After repeated administration, steady-state was achieved by day 5. The absorption rate of the SR formulation was lower and the 24 h peak-to-trough fluctuation was 4-fold lower compared to the IR formulation. After dose correction there wasno change in AUC, (726 +/- 207 and 690 +/- 183 ng x mL(-1) x h for SR and IR, respectively). The elimination parameters (t(1,2z), Ae(tau), and CLr) remained unchanged. The SR formulation showed sustained release of indapamide with a reduction in peak concentration, while steady-state level was not affected by formulations. The two formulations have the same bioavailability. PMID- 10796063 TI - Gastroduodenal tolerability of medium dose enteric-coated aspirin: a placebo controlled endoscopic study of a new enteric-coated formulation versus regular formulation in healthy volunteers. AB - We compared, in a cross-over study, the toxicity of 300 mg enteric-coated aspirin with regular aspirin used for the prevention of cardiovascular events. In terms of endoscopic haemorrhagic lesions, enteric-coated aspirin is less gastrotoxic than regular aspirin. PMID- 10796062 TI - Monitoring of the intracellular activation of 5-fluorouracil to deoxyribonucleotides in HT29 human colon cell line: application to modulation of metabolism and cytotoxicity study. AB - An HPLC method was developed for in vitro detection and monitoring of intracellular metabolites of [3H]-5-fluorouracil (FUra). Results showed a preferential activation of FUra to ribonucleoside and ribonucleotide derivatives (FURd, FUMP, FUDP and FUTP) in the human colorectal HT29 cell line. We screened various agents so as to determine if they could act as modulators of metabolism and/or toxicity of FUra by reversing the activation pathway of FUra from ribo- to deoxyribonucleotides, thus enhancing FdUMP formation. Different drugs (efflux inhibitors, catabolism inhibitors and enzymatic cofactors) were tested for enhancement of cytotoxicity when associated with FUra. The most promising agents were further studied by assessment of their ability to modulate intracellular activation of FUra to enhance thymidylate synthase (TS) inhibition by FUra and to increase the subsequent induction of apoptosis. 2'-Deoxyinosine (d-Ino), a deoxyribose 1-phosphate donor increasing thymidine phosphorylase activity, stood out as the best modulating agent we screened. Results showed an up to 30-fold increase of cytotoxicity along with a stronger inhibition of TS when FUra was associated with d-Ino, while FUra alone exhibited a lesser effect on TS activity. Besides, HPLC analysis revealed a complete reversal of the activation pathway of FUra, thus leading to an intracellular accumulation of deoxyribonucleotides. Assessment of cell cycle distribution showed a marked increase (+480%) of apoptosis in cells exposed to FUra/d-Ino compared to FUra alone. The HPLC method we developed is a convenient tool for assessing to what extent modulators will actually act on the intracellular activation of FUra. This study confirms the potentiality of d-Ino to modulate FUra metabolism in vitro. It proved to be an agent able to orientate the mechanism of action of FUra towards the inhibition of TS in cells where the normal activation pathway of the drug does not result in the intracellular accumulation of the active metabolite FdUMP. PMID- 10796064 TI - Lubeluzole-induced prolongation of cardiac action potential in rabbit Purkinje fibres. AB - Lubeluzole, a novel neuroprotective compound, has been associated with cases of QT interval prolongation but its effects on the cardiac action potential have not been described to date. Thus, the electrophysiological effects of lubeluzole were studied in rabbit isolated Purkinje fibres. The results demonstrate that lubeluzole (0.001-1 microM) concentration-dependently lengthened action potential duration at 50% and 90% of repolarization (APD50 and APD90) without significantly modifying other parameters. Furthermore, APD lengthening induced by lubeluzole was not significantly decreased by reducing the basic cycle length (from 3,000 to 1,000 ms). The results demonstrate that lubeluzole potently and concentration dependently increases APD from 0.01 microM, consistent with class III-type antiarrhythmic actions, which is likely to underlie QT interval prolongation induced by the drug. PMID- 10796065 TI - Imipenem but not meropenem induces convulsions in DBA/2 mice, unrelated to cerebrospinal fluid concentrations. AB - Imipenem and meropenem CSF diffusion was comparable in DBA/2 mice but only imipenem induced convulsions, not related to CSF concentration. PMID- 10796066 TI - Cyclic nucleotide phosphodiesterase (PDE) inhibitors and immunomodulation. AB - Intracellular levels of cyclic nucleotide second messengers are regulated predominantly by the complex superfamily of cyclic nucleotide phosphodiesterase (PDE) enzymes. Recent advances in our understanding of the molecular pharmacology of these enzymes has led to their identification as biologic regulators of certain disease states and the development of isozyme-selective inhibitors as potential therapeutic agents. A large body of in vitro and preclinical data suggests the therapeutic utility of PDE4 inhibitors as potent anti-inflammatory agents. Early clinical trials with selective PDE inhibitors substantiate this approach while highlighting pharmacodynamic and toxicologic pitfalls inherent to the inhibition of specific PDE isozymes. This commentary will review our current understanding of PDE inhibitors as immunomodulatory agents. PMID- 10796067 TI - Receptors mediating adenovirus attachment and internalization. AB - Adenovirus infection requires that the virus attach to cells and be internalized. Interaction between the viral fiber protein and specific cell surface receptors, such as the 46-kDa coxsackievirus and adenovirus receptor (CAR), is responsible for attachment; a second interaction between the viral penton base and cell surface integrins facilitates virus internalization. Expression of receptors may determine whether tissues are susceptible to adenovirus infection and adenovirus mediated gene delivery. PMID- 10796068 TI - Membrane interaction of an antitumor antibiotic, mithramycin, with anionic phospholipid vesicles. AB - Small unilamellar vesicles (SUV) composed of zwitterionic phosphatidylcholine and two different anionic phospholipids, phosphatidic acid and phosphatidylserine, in different compositions, were employed to study the membrane interaction of an antitumor antibiotic, mithramycin (MTR). Binding of MTR to dimyristoylphosphatidylcholine (DMPC) liposomes containing the anionic phospholipid dimyristoylphosphatidic acid (DMPA) was estimated by measuring the increase in intensity of the intrinsic fluorescence of MTR with increasing concentrations of phospholipids. Membrane perturbations were observed in acidic SUV of DMPC/DMPA and DMPC/bovine brain phosphatidylserine by MTR and its magnesium complex as studied by monitoring the leakage of the entrapped fluorescent marker carboxyfluorescein and by electron microscopic measurements of the size of the liposomes. These results indicated a possible role of anionic phospholipids in mediating binding of MTR and its magnesium complex to the cell surface membranes before reaching the target DNA. PMID- 10796069 TI - tert-butyl hydroperoxide/hemoglobin-induced oxidative stress and damage to vascular smooth muscle cells: different effects of nitric oxide and nitrosothiols. AB - The goal of the present work was to determine whether nitric oxide (NO) released from different donors (NONOates and nitrosothiols) can act as a protective antioxidant against oxidative stress and cytotoxicity induced by extracellular hemoglobin/tert-butyl hydroperoxide (Hb/tert-BuOOH) in vascular smooth muscle cells (VSMCs). No changes in phospholipid composition were found in VSMCs incubated with oxyhemoglobin (oxyHb)/tert-BuOOH. Using our newly developed HPLC fluorescence technique for measurement of site-specific oxidative stress in membrane phospholipids, we produced VSMCs in which endogenous phospholipids were metabolically labeled with an oxidation-sensitive fluorescent fatty acid, cis parinaric acid. In these cells, we were able to reliably quantitate oxidative stress in major phospholipid classes-phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, and phosphatidylinositol-induced by tert BuOOH in the presence of oxyHb or methemoglobin (metHb). The oxidative stress was accompanied by cytotoxic effects of oxyHb/tert-BuOOH and metHb/tert-BuOOH on VSMCs. We further found that an NO donor, (Z)-1-[N-(3-ammoniopropyl)-N-(n propyl)amino]diazen 1-ium-1,2-diolate (PAPANONO), but not nitrosothiols, protected VSMCs against oxidative stress and cytotoxicity induced by Hb/tert BuOOH. The protective effect of PAPANONO was most likely due to its ability to form NO-heme Hb (detectable by low temperature EPR spectroscopy and visible spectrophotometry). These findings are important for further understanding the physiological antioxidant role of NO against oxidative stress induced by hemoproteins as well as for pathological hypertensive events induced by extracellular Hb via NO depletion. PMID- 10796070 TI - Pharmacological characterization of glucocorticoid receptors in primary human bronchial epithelial cells. AB - Bronchial epithelial cells play an important role in amplifying and perpetuating airway inflammation and may be a target for inhaled steroids. We have characterized glucocorticoid receptors in primary human bronchial epithelial cells. Northern and western blot analyses demonstrated the expression of glucocorticoid receptor mRNA and protein, respectively, in primary bronchial epithelial cells. The activity of these receptors was shown using a radioligand binding assay. High-affinity binding with pharmacological specificity was demonstrated for [3H]dexamethasone. The equilibrium dissociation constant (Kd) and density of binding sites (Bmax) for [3H]dexamethasone determined from saturation isotherms were 4.4 nM x/divided by 0.95 (SEM) and 30.1 fmol/mg protein +/-6.4 (SEM). Glucocorticoid receptors were activated by dexamethasone as assessed using a glucocorticoid-responsive reporter plasmid, pTAT3-CAT. Transfection of primary human bronchial epithelial cells with this reporter plasmid resulted in 35-fold activation of transcription following dexamethasone stimulation (10(-6) M). The glucocorticoid receptor antagonist RU-486 (mifepristone) significantly counteracted the effect of dexamethasone on glucocorticoid receptor activation, indicating that the dexamethasone effect is specific and is mediated through the glucocorticoid receptor. In summary, our study demonstrated that primary cultures of human bronchial epithelial cells possess glucocorticoid receptors that function as a ligand-activated transcriptional regulator. The presence of glucocorticoid receptors confers their responsiveness to glucocorticoids and indicates that the airway epithelium may be a target for the anti-inflammatory effects of inhaled steroids. PMID- 10796071 TI - Mitochondrial 3-hydroxy-3-methylglutaryl coenzyme A synthase and carnitine palmitoyltransferase II as potential control sites for ketogenesis during mitochondrion and peroxisome proliferation. AB - 3-Thia fatty acids are potent hypolipidemic fatty acid derivatives and mitochondrion and peroxisome proliferators. Administration of 3-thia fatty acids to rats was followed by significantly increased levels of plasma ketone bodies, whereas the levels of plasma non-esterified fatty acids decreased. The hepatic mRNA levels of fatty acid binding protein and formation of acid-soluble products, using both palmitoyl-CoA and palmitoyl-L-carnitine as substrates, were increased. Hepatic mitochondrial carnitine palmitoyltransferase (CPT) -II and 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) synthase activities, immunodetectable proteins, and mRNA levels increased in parallel. In contrast, the mitochondrial CPT-I mRNA levels were unchanged and CPT-I enzyme activity was slightly reduced in the liver. The CoA ester of the monocarboxylic 3-thia fatty acid, tetradecylthioacetic acid, which accumulates in the liver after administration, inhibited the CPT-I activity in vitro, but not that of CPT-II. Acetoacetyl-CoA thiolase and HMG-CoA lyase activities involved in ketogenesis were increased, whereas the citrate synthase activity was decreased. The present data suggest that 3-thia fatty acids increase both the transport of fatty acids into the mitochondria and the capacity of the beta-oxidation process. Under these conditions, the regulation of ketogenesis may be shifted to step(s) beyond CPT-I. This opens the possibility that mitochondrial HMG-CoA synthase and CPT-II retain some control of ketone body formation. PMID- 10796072 TI - Protective effect of docosahexaenoic acid against hydrogen peroxide-induced oxidative stress in human lymphocytes. AB - Oxidatively stressed lymphocytes exhibit decreased proliferative response to mitogenic stimulation. Although several sensitive targets involved in lymphocyte suppression have already been identified, little is known about the influence of oxidative stress on cyclic nucleotide phosphodiesterases (PDE) (EC 3.1.4.17), thought to play a major role in the control of cyclic AMP (cAMP) level, a well recognized negative effector of lymphoproliferation. Although the polyunsaturated fatty acid content of membrane phospholipids is thought to be directly related to the extent of oxidant-induced lipid peroxidation, some n-3 fatty acids also seem to have antioxidant effects, depending on the concentration used and the overall redox status of the cells in question. Results of the present study showed that human peripheral blood mononuclear cells (PBMC) as well as rat thymocytes were relatively resistant to a short-term exposure (10 min) to hydrogen peroxide (H2O2). Indeed, H2O2-induced lipid peroxidation, estimated by malondialdehyde (MDA) production, was only 2-fold increased by H2O2 concentrations lower than 2 mM, whereas a larger increase (10-fold) could be observed in PBMC at the highest dose (5 mM). Previous enrichment of PBMC with 5 microM docosahexaenoic acid (22:6n-3), brought to the cells as a fatty acid-albumin complex (ratio 1), significantly reduced MDA production induced by low doses of H2O2, the protective effect no longer being observed at the highest doses. In contrast, eicosapentaenoic acid (20:5n-3) did not have any protective effect. Cytosolic PDE activities of both human PBMC and rat thymocytes were significantly inhibited (40 50%) after H2O2 treatment of the cells, whereas particulate PDE activities were not modified. Different responses of PDE activities to H2O2 treatment were observed when PBMC were first enriched with 22:6n-3 prior to H2O2 addition. In 22:6n-3-treated cells, the H2O2-induced inhibition of both cAMP- and cGMP-PDE cytosolic activities was abolished, whereas the particulate activities were increased by the highest H2O2 concentration used (5 mM). At the same time, the glutathione peroxidase (glutathione: oxidoreductase, EC 1.11.1.9) (GSH-Px) activity of PBMC and thymocytes was only marginally inhibited by H2O2 addition (20%), and pretreatment of the cells with 22:6n-3 did not modify the slight inhibitory effect of H2O2. Collectively, these results suggest that lymphocytes are relatively resistant to H2O2-induced lipid peroxidation due to their high GSH Px content, and that low doses of 22:6n-3 are able to prevent some of the H2O2 induced alterations such as lipid peroxidation and PDE inhibition. Docosahexaenoic acid might thus offer some protection against oxidant-induced lymphocyte suppression. PMID- 10796073 TI - Competition between iron(III)-selective chelators and zinc-finger domains for zinc(II). AB - Many iron(III)-selective chelators possess an appreciable affinity for zinc(II) and this can prove to be undesirable when such chelators are being assessed for clinical application. At present, there is no useful test available which can reliably access this problem. In the present manuscript, we provide evidence that indicates that a zinc-finger protein MTF-1, (metal transcription factor-1) may prove to be a suitable candidate. N,N',2-hydroxybenzyl ethylenediamine diacetic acid, in contrast to desferrioxamine, removes zinc quite efficiently from MTF-1. PMID- 10796074 TI - pH and drug resistance. I. Functional expression of plasmalemmal V-type H+-ATPase in drug-resistant human breast carcinoma cell lines. AB - A major obstacle for the effective treatment of cancer is the phenomenon of multidrug resistance (MDR) exhibited by many tumor cells. Many, but not all, MDR cells exhibit membrane-associated P-glycoprotein (P-gp), a drug efflux pump. However, most mechanisms of MDR are complex, employing P-gp in combination with other, ill-defined activities. Altered cytosolic pH (pHi) has been implicated to play a role in drug resistance. In the current study, we investigated mechanisms of pHi regulation in drug-sensitive (MCF-7/S) and drug-resistant human breast cancer cells. Of the drug-resistant lines, one contained P-gp (MCF-7/DOX; also referred to as MCF-7/D40) and one did not (MCF-7/MITOX). The resting steady-state pHi was similar in the three cell lines. In addition, in all the cell lines, HCO3 slightly acidified pHi and increased the rates of pHi recovery after an acid load, indicating the presence of anion exchanger (AE) activity. These data indicate that neither Na+/H+ exchange nor AE is differentially expressed in these cell lines. The presence of plasma membrane vacuolar-type H+-ATPase (pmV-ATPase) activity in these cell lines was then investigated. In the absence of Na+ and HCO3-, MCF-7/S cells did not recover from acid loads, whereas MCF-7/MITOX and MCF 7/DOX cells did. Furthermore, recovery of pHi was inhibited by bafilomycin A1 and NBD-Cl, potent V-ATPase inhibitors. Attempts to localize V-ATPase immunocytochemically at the plasma membranes of these cells were unsuccessful, indicating that V-ATPase is not statically resident at the plasma membrane. Consistent with this was the observation that release of endosomally trapped dextran was more rapid in the drug-resistant, compared with the drug-sensitive cells. Furthermore, the drug-resistant cells entrapped doxorubicin into intracellular vesicles whereas the drug-sensitive cells did not. Hence, it is hypothesized that the measured pmV-ATPase activity in the drug-resistant cells is a consequence of rapid endomembrane turnover. The potential impact of this behavior on drug resistance is examined in a companion manuscript. PMID- 10796076 TI - Effects of selected histamine H3 receptor antagonists on tele-methylhistamine levels in rat cerebral cortex. AB - The H3 antagonist thioperamide is thought to act on brain H3 autoreceptors to increase both the release and metabolism of neuronal histamine (HA). Our studies investigated the effects of several new brain-penetrating H3 antagonists on rat cerebral cortical levels of the HA metabolite tele-methylhistamine (t-MH). Animals were pretreated with H3 antagonists (0.3 to 30 mg/kg; 1-4 hr; i.p.) in the presence or absence of the monoamine oxidase inhibitor pargyline to prevent metabolism of t-MH. Cortical t-MH levels were measured by both radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS). Pargyline (60 mg/kg; 1 hr; i.p.) produced an approximately 2-fold increase in t-MH levels as measured by either GC-MS or RIA. Thioperamide (+/- pargyline) increased t-MH levels as measured by both GC-MS and RIA. In contrast, neither 5-cyclohexyl-1-(4-imidazol-4 ylpiperidyl)pentan-1-one (GT-2016) (+/- pargyline), 4-(6-cyclohexylhex-cis-3 enyl)imidazole (GT-2227) (+/- pargyline), nor clobenpropit (minus pargyline) increased t-MH levels as measured by GC-MS. A good agreement was found between t MH levels as determined by either RIA or GC-MS except after treatment with GT 2016, which increased apparent t-MH brain levels according to the former but not the latter method. Subsequent studies suggest the in vivo formation of a GT-2016 metabolite, which can cross-react in the t-MH RIA. Although all H3 receptor antagonists studied to date seem capable of enhancing brain HA release, only thioperamide presently was found to enhance cortical t-MH levels. Thus, H3 receptor antagonists may differentially affect HA release and turnover, and brain t-MH levels may not be reliable predictors of H3 agonist, partial agonist, or antagonist in vivo activity. PMID- 10796075 TI - pH and drug resistance. II. Turnover of acidic vesicles and resistance to weakly basic chemotherapeutic drugs. AB - Resistance to chemotherapeutic agents is a major cause of treatment failure in patients with cancer. The primary mechanism leading to a multidrug-resistant phenotype is assumed to be plasma-membrane localized overexpression of drug efflux transporters, such as P-glycoprotein (P-gp). However, acidic intracellular organelles can also participate in resistance to chemotherapeutic drugs. In this study, we investigated, both experimentally and theoretically, the effect of acidic vesicle turnover on drug resistance. We have developed a general model to account for multiple mechanisms of resistance to weakly basic organic cations, e.g. anthracyclines and Vinca alkaloids. The model predicts that lower cytosolic concentrations of drugs can be achieved through a combination of high endosomal turnover rates, a low endosomal pH, and an alkaline-inside pH gradient between cytosol and the extracellular fluid. Measured values for these parameters have been inserted into the model. Computations using conservative values of all parameters indicate that turnover of acidic vesicles can be an important contributor to the drug-resistant phenotype, especially if vesicles contain an active uptake system, such as H+/cation exchange. Even conservative estimates of organic cation-proton antiport activity would be sufficient to make endosomal drug extrusion a potent mechanism of resistance to weakly basic drugs. The effectiveness of such a drug export mechanism would be comparable to drug extrusion via drug pumps such as P-gp. Thus, turnover of acidic vesicles can be an important factor in chemoresistance, especially in cells that do not overexpress plasma membrane-bound drug pumps like P-glycoprotein. PMID- 10796077 TI - Sex-dependent induction of alcohol dehydrogenase activity in rats. AB - The glycolethers 2-methoxyethanol (2-ME), 2-ethoxyethanol (2-EE), and 2 butoxyethanol are widely used organic solvents with teratogenic, spermatotoxic, and hematotoxic effects due to the respective alkoxyacetic acid metabolites formed via alcohol dehydrogenase (ADH). ADH displays sexually dimorphic activities in adult rats, and is probably at least in part under the control of testosterone. The aim of this study was to investigate whether induction of ADH is also sex-dependent. Ethanol, 2-ME, and 2-EE were tested as inducers of hepatic and gastric ADH in female, male, and castrated male rats. The activity of hepatic ADH was higher in female than in male rats, while the activity of gastric ADH was higher in male than in female rats. The activities of ADH increased with increasing chain length of the glycolethers and alcohols. Castration of male rats led to a female pattern of ADH activity, i.e. increased activity of hepatic ADH and decreased activity of gastric ADH. Ethanol had no inducing effect on hepatic ADH in either male or female rats. 2-ME and 2-EE caused an increase in the activity of hepatic ADH in male and castrated male rats only. The present data demonstrate a different expression of ADH isoenzymes in male and female rats, and a sex-dependent induction of ADH isoenzymes. The different possible regulatory mechanisms for the different ADH isoenzymes require further investigation. PMID- 10796078 TI - Positive regulation of the rat CYP2B2 phenobarbital response unit by the nuclear receptor hexamer half-site.nuclear factor 1 complex. AB - A distal 163-bp fragment mediates phenobarbital responsiveness of the rat CYP2B2 gene. Multiple cis-acting elements in this fragment cooperate to form a phenobarbital response unit (PBRU). A nuclear factor 1 binding site and an associated nuclear receptor hexamer half-site are present in both the rat CYP2B2 PBRU and the homologous mouse Cyp2b10 sequence. Based on mutational analyses, the hexamer half-site has been reported to act positively in CYP2B2 and negatively in Cyp2b10. However, the specific mutations introduced into the rat and mouse hexamer half-sites were different, raising the possibility that the different roles attributed to the element may be a consequence of the different mutations used. We introduced into the rat CYP2B2 hexamer half-site the specific mutational change previously introduced into the Cyp2b10 sequence, where its effect was to increase the basal level of expression and to abolish phenobarbital responsiveness. In the rat context, this mutation reduced but did not abolish phenobarbital responsiveness and decreased, rather than increased, the basal level of expression. The residual phenobarbital responsiveness of the hexamer half-site mutant, as well as that of nuclear factor 1 mutants, indicates that these elements behave as positive accessory sites, suggesting that factors binding to them function as activators of phenobarbital-dependent transcription. PMID- 10796079 TI - Abdominal decompression for suspected fetal compromise/pre-eclampsia. AB - BACKGROUND: Abdominal decompression was developed as a means of pain relief during labour. It has also been used for complications of pregnancy, and in healthy pregnant women in an attempt to improve fetal wellbeing and intellectual development. OBJECTIVES: The objective of this review was to assess the effects of antenatal abdominal decompression for maternal hypertension or impaired fetal growth, on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1998. SELECTION CRITERIA: Randomised or quasi-randomised trials comparing abdominal decompression with no decompression in women with pre-eclampsia and/or fetuses thought to be compromised. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Three studies were included, all with the possibility of containing serious bias. Therapeutic abdominal decompression was associated with the following reductions: persistent pre-eclampsia (relative risk 0.36, 95% confidence interval 0.18 to 0.72); fetal distress in labour (relative risk 0.37, 95% confidence interval 0.19 to 0.71); low birthweight (relative risk 0.50, 95% confidence interval 0.40 to 0. 63); Apgar scores less than six at one minute (relative risk 0.26, 95% confidence interval 0.12 to 0.56); and perinatal mortality (relative risk 0.39, 95% confidence interval 0.22 to 0.71). REVIEWER'S CONCLUSIONS: Due to the methodological limitations of the studies, the effects of therapeutic abdominal decompression are not clear. The apparent improvements in birthweight and perinatal mortality warrant further evaluation of abdominal decompression where there is impaired fetal growth and possibly for women with pre-eclampsia. PMID- 10796080 TI - Absorbable staples for uterine incision at caesarean section. AB - BACKGROUND: Staples can be placed during the making of an incision, with the aim of decreasing blood loss from the cut edges. OBJECTIVES: The objective of this review was to assess the effects of using a stapler with absorbable staples to extend the uterine incision at lower segment caesarean section. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of extending the uterine incision using a stapler compared with extending the incision digitally or with scissors in women having a lower segment transverse incision caesarean section. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. MAIN RESULTS: Four trials involving 526 women were included. There was no difference in total operating time between the stapling technique and other techniques to extend the incision (weighted mean difference -1.17 minutes, 95% confidence interval -3.57 to 1.22). However stapling devices increased the time needed to deliver the baby (weighted mean difference 0.85 minutes, 95% confidence interval 0.48 to 1.23). Blood loss was lower with the use of staples (weighted mean difference -41.22 millilitres, 95% confidence interval -50.63 to -31.8). No significant differences between stapling and other techniques were detected for other perinatal morbidity outcomes. REVIEWER'S CONCLUSIONS: There is not enough evidence to justify the routine use of stapling devices to extend the uterine incision at lower segment caesarean section. There is a possibility that stapling could cause harm, by prolonging the time to deliver the baby. PMID- 10796081 TI - Absorbable synthetic versus catgut suture material for perineal repair. AB - BACKGROUND: Approximately 70% of women will experience some degree of perineal trauma following vaginal delivery and will require stitches. This may result in perineal pain and superficial dyspareunia. OBJECTIVES: The objective of this review was to assess the effects of absorbable synthetic suture material as compared with catgut on the amount of short and long term pain experienced by mothers following perineal repair. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials comparing absorbable synthetic (polyglycolic acid and polyglactin) with plain or chromic catgut suture for perineal repair in mothers after vaginal delivery. DATA COLLECTION AND ANALYSIS: Trial quality was assessed independently by two reviewers. Data were extracted by one reviewer and checked by the second reviewer. MAIN RESULTS: Eight trials were included. Compared with catgut, the polyglycolic acid and polyglactin groups were associated with less pain in first three days (odds ratio 0.62, 95% confidence interval 0.54 to 0.71). There was also less need for analgesia (odds ratio 0.63, 95% confidence interval 0.52 to 0.77) and less suture dehiscence (odds ratio 0.45, 95% confidence interval 0.29 to 0.70). There was no significant difference in long term pain (odds ratio 0.81, 95% confidence interval 0.61 to 1.08). Removal of suture material was significantly more common in the polyglycolic acid and polyglactin groups (odds ratio 2.01, 95% confidence interval 1.56 to 2.58). There was no difference in the amount of dyspareunia experienced by women. REVIEWER'S CONCLUSIONS: Absorbable synthetic suture material (in the form of polyglycolic acid and polyglactin sutures) for perineal repair following childbirth appears to decrease women's experience of short-term pain. The length of time taken for the synthetic material to be absorbed is of concern. A trial addressing the use of polyglactin has recently been completed and this has been included in this updated review. PMID- 10796082 TI - Active versus expectant management in the third stage of labour. AB - BACKGROUND: Expectant management of the third stage of labour involves allowing the placenta to deliver spontaneously or aiding by gravity or nipple stimulation. Active management involves administration of a prophylactic oxytocic after delivery, early cord clamping and cutting, and controlled cord traction of the umbilical cord. OBJECTIVES: The objective of this review was to assess the effects of active versus expectant management on blood loss, post partum haemorrhage and other maternal and perinatal complications of the third stage of labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials comparing active and expectant management of the third stage of labour in women with singleton pregnancies whose babies were presenting head first and who were expecting a vaginal delivery. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by the reviewers. MAIN RESULTS: Four studies were included. Three of the trials were of good quality. Compared to expectant management, active management (in the setting of a maternity hospital) was associated with the following reduced risks: maternal blood loss (weighted mean difference -79.33 millilitres, 95% confidence interval -94.29 to -64. 37); post partum haemorrhage of more than 500 millilitres (odds ratio 0.34, 95% confidence interval 0.28 to 0.41); prolonged third stage of labour (weighted mean difference -3.40 minutes, 95% confidence interval -4.66 to -2.13). Active management was associated with an increased risk of maternal nausea (odds ratio 1. 95, 95% confidence interval 1.58 to 2.42), vomiting and raised blood pressure (probably due to the use of ergometrine). No advantages or disadvantages were apparent for the baby. REVIEWER'S CONCLUSIONS: Routine 'active management' is superior to 'expectant management' in terms of blood loss, post partum haemorrhage and other serious complications of the third stage of labour. Active managment is, however, associated with an increased risk of unpleasant side effects (eg nausea and vomiting), and hypertension, where ergometrine is used. Active management should be the routine management of choice for women expecting a single baby by vaginal delivery in a maternity hospital. The implications are less clear for other settings including domiciliary practice (in developing and industrialised countries). PMID- 10796083 TI - Home-like versus conventional institutional settings for birth. AB - BACKGROUND: Many home-like birth centres have been established near conventional labour wards for the care of pregnant women who prefer and require little or no medical intervention during labour and birth. OBJECTIVES: The objective of this review was to assess the effects of care in a home-like birth environment compared to care in a conventional labour ward, on labour and birth outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing a home-like institutional birth environment to conventional hospital care for pregnant women at low risk of obstetric complications. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Five trials involving almost 8000 women were included. Substantial numbers of women allocated to home-like settings were transferred to standard care before or during labour, making interpretation of results difficult. Allocation to a home-like setting was associated with lower rates of intrapartum analgesia/anaesthesia (odds ratio 0.82, 95% confidence interval 0.72 to 0.93), fetal heart rate abnormalities (0.72, 95% confidence interval 0.63 to 0.81), augmented labour, and immobility during labour, as well as greater satisfaction with care, and increased likelihood of sore nipples and mastitis. There was a non-statistically significant trend towards higher perinatal mortality in the home-like setting (odds ratio 1.49, 95% confidence interval 0.79 to 2.78). REVIEWER'S CONCLUSIONS: There appear to be some benefits from home-like settings for childbirth, although increased support from caregivers may be more important. Caregivers and clients in home-like settings need to watch for signs of complications. PMID- 10796084 TI - Amnioinfusion for umbilical cord compression in labour. AB - BACKGROUND: Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. OBJECTIVES: The objective of this review was to assess the effects of amnioinfusion on maternal and perinatal outcome for potential or suspected umbilical cord compression or potential amnionitis. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials of amnioinfusion compared with no amnioinfusion in women with babies at risk of umbilical cord compression; and women at risk of intrauterine infection. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: Twelve studies were included. Transcervical amnioinfusion for potential or suspected umbilical cord compression was associated with the following reductions: fetal heart rate decelerations (relative risk 0.54, 95% confidence interval 0.43 to 0.68); caesarean section for suspected fetal distress (relative risk 0.35, 95% confidence interval 0.24 to 0.52); neonatal hospital stay greater than 3 days (relative risk 0.40, 95% confidence interval 0. 26 to 0.62); maternal hospital stay greater than 3 days (relative risk 0.46, 95% 0.29 to 0.74). Transabdominal amnioinfusion showed similar results. Transcervical amnioinfusion to prevent infection in women with membranes ruptured for more than 6 hours was associated with a reduction in puerperal infection (relative risk 0.50, 95% confidence interval 0.26 to 0.97). REVIEWER'S CONCLUSIONS: Amnioinfusion appears to reduce the occurrence of variable heart rate decelerations and lower the use of caesarean section. However the studies were done in settings where fetal distress was not confirmed by fetal blood sampling. The results may therefore only be relevant where caesarean sections are commonly done for abnormal fetal heart rate alone. The trials reviewed are too small to address the possibility of rare but serious maternal adverse effects of amnioinfusion. PMID- 10796085 TI - Amnioinfusion for meconium-stained liquor in labour. AB - BACKGROUND: Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. It is also thought to dilute meconium when present in the amniotic fluid and so reduce the risk of meconium aspiration. However it may be that the mechanism of effect is that it corrects oligohydramnios (reduced amniotic fluid), for which thick meconium staining is a marker. OBJECTIVES: The objective of this review was to assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium-staining of the amniotic fluid. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Ten studies, most involving small numbers of participants, were included. Under standard perinatal surveillance, amnioinfusion was associated with a reduction in the following: heavy meconium staining of the liquor (relative risk 0.03, 95% confidence interval 0.01 to 0.15); variable fetal heart rate deceleration (relative risk 0.47, 95% confidence interval 0.24 to 0. 90); and a trend to reduced caesarean section overall (relative risk 0.83, 95% confidence interval 0.69 to 1.00). No perinatal deaths were reported. Under limited perinatal surveillance, amnioinfusion was associated with a reduction in the following: meconium aspiration syndrome (relative risk 0.24, 95% confidence interval 0. 12 to 0.48); neonatal hypoxic ischaemic encephalopathy (relative risk 0.07, 95% confidence interval 0.01 to 0.56) and neonatal ventilation or intensive care unit admission (relative risk 0.56, 95% confidence interval 0.39 to 0.79); there was a trend towards reduced perinatal mortality (relative risk 0.34, 95% confidence interval 0.11 to 1.06). REVIEWER'S CONCLUSIONS: Amnioinfusion is associated with improvements in perinatal outcome, particularly in settings where facilities for perinatal surveillance are limited. The trials reviewed are too small to address the possibility of rare but serious maternal adverse effects of amnioinfusion. PMID- 10796086 TI - Amniotomy for shortening spontaneous labour. AB - BACKGROUND: Early amniotomy has been advocated as a component of the active management of labour. Several randomised trials comparing routine amniotomy to an attempt to conserve the membranes have been published. Their limited sample sizes limit their ability to address the effects of amniotomy on indicators of maternal and neonatal morbidity. OBJECTIVES: To study the effects of amniotomy on the rate of Cesarean delivery and on other indicators of maternal and neonatal morbidity (Apgar less than 7 at 5 minutes, admission to NICU). SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled trials of amniotomy during first stage of labour were eligible. DATA COLLECTION AND ANALYSIS: Data were extracted by two trained reviewers from published reports. Trials were assigned methodological quality scores based on a standardized rating system. Typical odds ratios (ORs) were calculated using Peto's method. MAIN RESULTS: Amniotomy was associated with a reduction in labour duration of between 60 and 120 minutes. There was a marked trend toward an increase in the risk of Cesarean delivery: OR = 1.26; 95% Confidence Interval (CI)=0.96-1. 66. The likelihood of a 5 minute Apgar score less than 7 was reduced in association with early amniotomy (OR = 0.54; 95% CI = 0.30-0.96). Groups were similar with respect to other indicators of neonatal status (arterial cord pH, NICU admissions). There was a statistically significant association of amniotomy with a decrease in the use of oxytocin: OR = 0.79; 95% CI = 0.67-0.92. REVIEWER'S CONCLUSIONS: Routine early amniotomy is associated with both benefits and risks. Benefits include a reduction in labour duration and a possible reduction in abnormal 5-minute Apgar scores. The meta-analysis provides no support for the hypothesis that routine early amniotomy reduces the risk of Cesarean delivery. Indeed there is a trend toward an increase in Cesarean section. An association between early amniotomy and Cesarean delivery for fetal distress is noted in one large trial. This suggests that amniotomy should be reserved for women with abnormal labour progress. PMID- 10796087 TI - Prenatal thyrotropin-releasing hormone for preterm birth. AB - BACKGROUND: Adding thyrotropin-releasing hormones (TRH) to corticosteroids has been suggested as a way of improving fetal lung development. OBJECTIVES: The objective of this review was to assess the effect of giving prenatal TRH in addition to corticosteroids to women at risk of very preterm birth for the prevention of neonatal respiratory disease. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, Cochrane Controlled Trials Register, and bibliographies. Date of last search: January 1999. SELECTION CRITERIA: Randomised controlled trials in women at sufficient risk of preterm birth to warrant the use of prenatal corticosteroids to promote lung maturity. TRH and corticosteroids were compared with corticosteroids with or without placebo. The main outcomes considered were fetal and neonatal mortality, neonatal morbidity, childhood development and maternal morbidity. DATA COLLECTION AND ANALYSIS: All assessments of trial eligibility, quality and data extractions were done by at least two authors independently. MAIN RESULTS: Over 4500 women were recruited into the 11 included trials. Five trials were rated of high quality. Overall, prenatal TRH, in addition to corticosteroids, did not reduce the risk of neonatal respiratory disease, chronic oxygen dependance, or improve fetal, neonatal or childhood outcome in any of the outcomes assessed by intention-to treat analyses. Indeed, the data showed prenatal TRH to have adverse effects for women and their infants. All side effects monitored were more likely to occur in women receiving TRH. In the infants, prenatal TRH increased the risk of infants needing ventilation (relative risk (RR) 1.16, 95% confidence interval (CI) 1.03 1.29), having a low Apgar score at five minutes (RR 1.80, 95% CI 1.14-1.92) and, for the one trial providing data, was associated with poorer outcomes at the 12 month follow up. Sensitivity analyses by trial quality, or subgroups with differing times from entry to delivery, or different dose regimens of TRH, did not change these findings. REVIEWER'S CONCLUSIONS: On the basis of currently available evidence, prenatal TRH, in addition to corticosteroids, given to women at risk of very preterm birth cannot be recommended for clinical practice. PMID- 10796088 TI - Anti-D administration in pregnancy for preventing rhesus alloimmunisation. AB - BACKGROUND: A woman may develop Rh-negative antibodies during her first pregnancy when her fetus is Rh-positive. Antibodies develop most frequently after the 28th week of gestation. OBJECTIVES: The objective of this review was to asses the effects of giving antenatal anti-D immunoglobulin at 28 weeks or more of pregnancy on the incidence of RhD alloimmunisation when given to Rhesus negative mothers without anti-D antibodies. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, Cochrane Controlled Trials Register, and bibliographies. Date of last search: December 1998. SELECTION CRITERIA: Randomised trials in Rhesus negative women without anti-D antibodies given anti-D after 28 weeks of pregnancy, compared with no treatment or placebo. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and double entered. MAIN RESULTS: Two eligible trials, which involved over 4500 women, compared anti-D prophylaxis with no treatment. Although the data suggested, when women receive anti-D at 28 and 34 weeks gestation, a reduced incidence of immunisation during pregnancy (0R O.44, 95% CI 0.18-1.12), after the birth of a Rhesus positive infant (OR 0.44, 95% CI 0.18-1.12), and within 12 months after birth of a Rhesus positive infant (OR 0.44, 95% CI 0.19-1.01), none of these differences were statistically significant. In the trial, which used the larger dose of anti-D (100ug; 500IU), there was a clear reduction in the incidence of immunisation at 2-12 months following birth in women who had received Anti-D at 28 and 34 weeks (OR 0.22 95% CI 0.05-0.88). No data were available for the risk of RhD alloimmunisation in a subsequent pregnancy. No differences were observed in the incidence of neonatal jaundice. REVIEWER'S CONCLUSIONS: The risk of RhD alloimmunisation during or immediately after a first pregnancy is about 1.5%. Administration of 100ug (500IU) anti-D at 28 weeks and 34 weeks gestation to women in their first pregnancy can reduce this risk to about 0.2% without, to date, any adverse effects. Although such a policy is unlikely to confer benefit or improve outcome in the present pregnancy, fewer women will have Rhesus D antibodies in their next pregnancy. Adoption of such a policy will need to consider the costs of prophylaxis against the costs of care for women who become sensitised and their affected infants, and local adequacy of supply of anti-D gammaglobulin. PMID- 10796089 TI - Anti-D administration after childbirth for preventing Rhesus alloimmunisation. AB - BACKGROUND: The development of Rh immunisation and its prophylactic use since the 1970s has meant that severe Rhesus D (RhD) alloimmunisation is now rarely seen. OBJECTIVES: The objective of this systematic review was to assess the effects of giving anti-D to Rhesus negative women, with no anti-D antibodies, who had given birth to a Rhesus positive infant. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, MEDLINE (from 1966 to January 1999) and reference lists of relevant articles. Date of last search of Cochrane Controlled Trials Register: January 1999. SELECTION CRITERIA: Randomised trials in Rhesus negative women without antibodies who were given anti-D immunoglobulin postpartum compared with no treatment or placebo. DATA COLLECTION AND ANALYSIS: Assessments of inclusion criteria, trial quality and data extraction were done by each author independently. Initial analyses included all trials. Other analyses assessed the effect of trial quality, ABO compatibility and dose. MAIN RESULTS: Six eligible trials compared postpartum anti-D prophylaxis with no treatment or placebo. The trials involved over 10,000 women, but trial quality varied. Anti-D lowered the incidence of RhD alloimmunisation six months after birth (relative risk 0.04, 95% confidence interval 0.02 to 0.06), and in a subsequent pregnancy (relative risk 0.12, 95% confidence interval 0. 07 to 0.23). These benefits were seen regardless of the ABO status of the mother and baby and when anti-D was given within 72 hours of birth. Higher doses (up to 200 micro grams) were more effective than lower doses (up to 50 micro grams) in preventing RhD alloimmunisation in a subsequent pregnancy. REVIEWER'S CONCLUSIONS: Anti-D, given within 72 hours after childbirth, reduces the risk of RhD alloimmunisation in Rhesus negative women who have given birth to a Rhesus positive infant. However the evidence on the optimal dose is limited. PMID- 10796090 TI - Anticonvulsants for women with pre-eclampsia. AB - BACKGROUND: Pre-eclampsia is a relatively common complication of pregnancy. Anticonvulsants are used in the belief they help prevent eclamptic fits and subsequent poor outcomes for mother and infant. OBJECTIVES: The objective of this review was to assess the effects of anticonvulsants for women with pre-eclampsia on the women and their children. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, and the Cochrane Controlled Trials Register, 1999 Issue 3. SELECTION CRITERIA: Randomised trials comparing anticonvulsants with placebo or no anticonvulsants or comparisons of different anticonvulsants in women with pre-eclampsia. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Nine studies were included. Comparing magnesium sulphate with placebo/no anticonvulsant the relative risk (RR) of eclampsia was 0.33, 95% confidence interval (CI) 0.11 to 1.02. There was no significant difference detected in the risk of caesarean section (RR 1.04, 95% CI 0.92 to 1.17). Magnesium sulphate appeared to be better than phenytoin at reducing the risk of eclampsia (RR 0.05, 95% CI 0.00 to 0.84). However there was an increased risk of caesarean section with magnesium sulphate compared to phenytoin (RR 1.21, 95% CI 1.05 to 1. 41). No statistically significant differences were reported for any other clinically important outcomes. Studies comparing magnesium sulphate and diazepam were too small for any reliable conclusions. REVIEWER'S CONCLUSIONS: There is not enough evidence to establish the benefits and hazards of anticonvulsants for women with pre-eclampsia. If an anticonvulsant is used, magnesium sulphate appears to be the best choice. PMID- 10796091 TI - Antihistamines versus aspirin for itching in late pregnancy. AB - BACKGROUND: While not common, itching in pregnancy (not due to liver disease) can be distressing. OBJECTIVES: The objective of this review was to assess the effects of treatment for itching in late pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. In addition, the Cochrane Controlled Trials Register (CENTRAL/CCTR) was searched. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials of treatments for itching in women in late pregnancy with normal liver function. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: One study of 38 women was included. This was a small crossover trial, using alternate allocation. The trial compared a histamine, chlorpheniramine, with aspirin. Aspirin was more effective than chlorpheniramine in relieving itching (odds ratio 2. 39, 95% confidence interval 1.25 to 4.57). However chlorpheniramine was more effective than aspirin when a rash was present. REVIEWER'S CONCLUSIONS: Aspirin appears to be more effective than chlorpheniramine for relief of itching in pregnancy when no rash is present. If there is a rash, chlorpheniramine may be more effective. PMID- 10796092 TI - Balanced protein/energy supplementation in pregnancy. AB - BACKGROUND: Observational and non-randomized studies have suggested that energy/protein supplementation in pregnant women increases gestational weight gain and fetal growth. OBJECTIVES: The objective of this review was to assess the effects of a balanced protein/energy supplement for pregnant women on gestational weight gain and on the outcome of pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Date of last search: January 2000. SELECTION CRITERIA: Acceptably controlled trials of energy/protein supplementation for pregnant women in which the protein content of the supplement was 'balanced' (protein content less than 25% of total energy content). DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Thirteen trials were included. They were of variable quality. Balanced protein/energy supplementation was associated with modest increases in maternal weight gain (weighted mean difference 17 grams per week, 95% confidence interval 5-29 grams per week) and fetal growth (birth weight increase, weighted mean difference 25 grams, 95% confidence interval 4-55 grams). The reduction in risk of small for gestational birth was substantial, however (odds ratio 0. 64, 95% confidence interval 0.53-0.78). These effects did not appear to be greater in undernourished women, nor did they seem to confer long term benefits to the child. No significant effects were detected on preterm birth, but significant reductions in stillbirth and neonatal death (based on only 3 trials) appear important. REVIEWER'S CONCLUSIONS: Balanced energy/protein supplementation improves fetal growth and may reduce the risk of fetal and neonatal death. The evidence is insufficient to evaluate whether there are other potential benefits to pregnant women or their infants. PMID- 10796093 TI - Bed rest in hospital for suspected impaired fetal growth. AB - BACKGROUND: Bed rest in hospital or at home is widely advised for many complications of pregnancy. The increased clinical supervision needs to be balanced with the risk of thrombosis, the stress on the pregnant women, as well as the costs to families and health services. OBJECTIVES: The objective of this review was to assess the effects of bed rest in hospital for women with suspected impaired fetal growth. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: December 1999. SELECTION CRITERIA: Randomised trials comparing a policy of bed rest in hospital with ambulatory management for women with suspected impaired fetal growth. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: One study involving 107 women was included. Allocation of treatment was by odd or even birth date. There were differences in baseline fetal weights and birth weights, but these were not statistically significant (mean estimated fetal weight deviation at enrolment was -21.7% for the bed rest group and -20.7% for the ambulatory group; mean estimated birth weight was -19.7% for the bed rest group and -20.6% for the ambulatory group). No differences were detected between bed rest and ambulatory management for fetal growth parameters (relative risk 0.43, 95% confidence interval: 0.15 to 1. 27) and neonatal outcomes. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of a bed rest in hospital policy for women with suspected impaired fetal growth. PMID- 10796094 TI - Tocolytics for suspected intrapartum fetal distress. AB - BACKGROUND: Prophylactic tocolysis with betamimetics and other agents has become widespread as a treatment for fetal distress. Uterine relaxation may improve placental blood flow and therefore fetal oxygenation. However there may also be adverse maternal cardiovascular effects. OBJECTIVES: The objective of this review was to assess the effects of tocolytic therapy for suspected fetal distress on fetal, maternal and perinatal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials comparing tocolytic therapy with no treatment or treatment with another tocolytic agent for suspected fetal distress. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Three studies were included. Compared with no treatment, there were fewer failed improvements in fetal heart rate abnormalities with tocolytic therapy (relative risk 0.26, 95% 0.13 to 0.53). Betamimetic therapy compared with magnesium sulphate showed a non significant trend towards reduced uterine activity (relative risk 0.07, 95% confidence interval 0.00 to 1.10). REVIEWER'S CONCLUSIONS: Betamimetic therapy appears to be able to reduce the number of fetal heart rate abnormalities and perhaps reduce uterine activity. However there is not enough evidence based on clinically important outcomes to evaluate the use of betamimetics for suspected fetal distress. PMID- 10796095 TI - Betamimetics for suspected impaired fetal growth. AB - BACKGROUND: Betamimetic drugs may promote fetal growth by increasing the availability of nutrients and by decreasing vascular resistance. They may also induce adverse effects via their effects on carbohydrate metabolism. OBJECTIVES: The objective of this review was to assess the effects of betamimetic therapy for suspected impaired fetal growth on fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of the latest search: December 1999. SELECTION CRITERIA: Randomised trials of betamimetic therapy compared with no betamimetic therapy or placebo in women with suspected impaired fetal growth. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed. MAIN RESULTS: Two studies of 118 women were included. No differences were found between the betamimetic groups and the control groups for low birth weight (relative risk 1.17, 95% confidence interval 0.75 to 1.83), other anthropometric or neonatal morbidity and mortality measures. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of betamimetics in promoting fetal growth. Larger studies are needed to investigate possible adverse effects of long term betamimetic administration. PMID- 10796096 TI - Tocolysis for preventing fetal distress in second stage of labour. AB - BACKGROUND: Prophylactic tocolysis with betamimetics and other agents has become widespread as a treatment for fetal distress. Uterine relaxation may improve placental blood flow and therefore fetal oxygenation. However there may also be adverse maternal cardiovascular effects. OBJECTIVES: The objective of this review was to assess the effects of prophylactic betamimetic therapy during the second stage of labour on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials comparing prophylactic intravenous betamimetic therapy during the second stage of labour with placebo or no treatment in uncomplicated pregnancies. DATA COLLECTION AND ANALYSIS: The reviewers assessed trial quality and extracted data. MAIN RESULTS: One study involving 100 women was included. Compared to placebo, prophylactic betamimetic therapy was associated with an increase in forceps deliveries (relative risk 1.83, 95% confidence interval 1.02 to 3. 29). There were no clear effects on postpartum haemorrhage, neonatal irritability, feeding slowness, umbilical arterial pH values or Apgar scores at 2 minutes. REVIEWER'S CONCLUSIONS: There is no evidence to support the prophylactic use of betamimetics during the second stage of labour. PMID- 10796097 TI - Biophysical profile for fetal assessment in high risk pregnancies. AB - BACKGROUND: Biophysical profile usually includes ultrasound monitoring of fetal movements, fetal tone and fetal breathing, ultrasound assessment of amniotic fluid volume and assessment of fetal heart rate by electronic monitoring. OBJECTIVES: The objective of this review was to assess the effects of biophysical profile tests on pregnancy outcome in high risk pregnancies. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register. Date of last search: October 1998. SELECTION CRITERIA: Randomised trials comparing fetal biophysical profile with other forms of fetal assessment in women with high risk pregnancies. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Four studies were included. Most trials were not of high quality. No difference was found between biophysical profile and other forms of fetal assessment over a range of fetal and neonatal measures. REVIEWER'S CONCLUSIONS: At present, there is not enough evidence from randomised trials to evaluate the use of biophysical profile as a test of fetal well-being in high risk pregnancies. PMID- 10796098 TI - Prophylactic versus selective blood transfusion for sickle cell anaemia during pregnancy. AB - BACKGROUND: Sickle cells have a shorter life span than normal red blood cells. It has been suggested that pregnancy complications for women with sickle cell anaemia may be reduced by regular blood transfusions. The aim is to maintain haemoglobin at 60-70% of the normal total. OBJECTIVES: The objective of this review was to assess the effects of a policy of routine blood transfusion for pregnant women with sickle cell disease with a policy of selective transfusion. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Acceptably controlled trials of blood transfusion in pregnant women with sickle cell disease. DATA COLLECTION AND ANALYSIS: Eligibility, trial quality assessment and data extraction were done by one reviewer. MAIN RESULTS: One trial involving 72 women was included. Half the women received blood transfusion only if haemoglobin fell below 6g% and the other half received two units of blood every week for three weeks, or until haemoglobin level was 10-11g%. A policy of selective transfusion reduced the number of transfusions required at the expense of more frequent pain crises. REVIEWER'S CONCLUSIONS: There is not enough evidence to draw conclusions about the prophylactic use of blood transfusion for sickle cell anaemia during pregnancy. PMID- 10796101 TI - Early versus delayed initiation of breastfeeding. AB - BACKGROUND: It has been suggested that the timing of a baby's first breastfeed may influence breastfeeding duration and emotional attachment. OBJECTIVES: The objective of this review was to assess the effects of breastfeeding soon after birth (within 30 minutes) compared to being breastfed later (between 4 to 8 hours after delivery) on the duration of breastfeeding and the mother/infant relationship. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing early skin contact and breastfeeding with late skin contact and breastfeeding in women intending to breastfeed their healthy term infant. DATA COLLECTION AND ANALYSIS: Data were extracted by two reviewers. MAIN RESULTS: Three studies involving 209 women were included. Compared with late contact and breastfeeding, early contact and breastfeeding was associated with greater communication between mother and infants in a two minute observation period (odds ratio 0.14, 95% confidence interval 0.03 to 0.61). There was no difference detected for numbers of women breastfeeding after birth (odds ratio for 12 weeks after birth 0.73, 95% confidence interval 0.34 to 1.54). REVIEWER'S CONCLUSIONS: No differences were found between early and delayed contact in regard to breastfeeding duration. Early contact was associated with greater communication between mothers and infants. PMID- 10796103 TI - Caesarean delivery for the second twin. AB - BACKGROUND: The optimal mode of delivery for a second twin in breech position is controversial, with support for both caesarean and vaginal delivery. OBJECTIVES: The objective of this review was to assess the effects of caesarean delivery compared with vaginal delivery of a second twin not presenting cephalically. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, Cochrane Controlled Trials Register, and bibliographies up to December 1998. SELECTION CRITERIA: Randomised trials of Caesarean delivery compared with vaginal delivery in women with a twin pregnancy where the second twin is not presenting cephalically. DATA COLLECTION AND ANALYSIS: All assessments of trial eligibility, quality and data extractions were done by the author without blinding. MAIN RESULTS: One trial involving 60 pairs of twins was included. Maternal febrile morbidity were increased in women allocated to the caesarean group (relative risk (RR) 3.67, 95% confidence interval (CI) 1.15-11.69), and there was a trend to an increased need for use of general anaesthesia (RR 2.40, 95% CI 0. 98-5.88). No differences were detected in neonatal outcome. REVIEWER'S CONCLUSIONS: caesarean section for delivery of a second twin not presenting cephalically is associated with increased maternal febrile morbidity with, as yet, no identified improvement in neonatal outcome. This policy should not be adopted except within the context of further controlled trials PMID- 10796104 TI - Calcium channel blockers for potential impaired fetal growth. AB - BACKGROUND: Calcium channel blockers may increase the blood flow to the fetus or may improve fetal-placental cellular energy generation. This could enhance fetal growth. OBJECTIVES: The objectives of this review were to assess the effects of calcium channel blockers on fetal growth and neonatal morbidity and mortality in pregnancies where impaired fetal growth is suspected. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of the last search: December 1999. SELECTION CRITERIA: Acceptably controlled trials of calcium channel blockers in women with potential impaired fetal growth. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. MAIN RESULTS: One study of 100 women (all smokers) was included. Mean birth weight was significantly higher in women receiving flunarizine compared to the control group. No other significant differences were found. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of calcium channel blockers for impaired fetal growth. The apparent beneficial effect of calcium channel blockers on birthweight warrants further investigation. PMID- 10796105 TI - Cephalic version by postural management for breech presentation. AB - BACKGROUND: It is possible that maternal posture may influence fetal position. Many postural techniques have been used to promote cephalic version. OBJECTIVES: The objective of this review was to assess the effects of postural management of breech presentation on measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth trials register and Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing postural management with pelvic elevation for breech presentation, with a control group. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by both reviewers. MAIN RESULTS: Three studies involving a total of 192 women were included. Postural management was associated with a non-significant trend towards fewer non-cephalic births (relative risk 0.90, 95% confidence interval 0. 73 to 1.12). No differences were detected for caesarean sections or Apgar scores below seven at one minute. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of postural management for breech presentation. PMID- 10796106 TI - Interventions for treating genital chlamydia trachomatis infection in pregnancy. AB - BACKGROUND: Chlamydia trachomatis is a sexually transmitted infection. Mother-to child transmission can occur at the time of birth and may result in ophthalmia neonatorum or pneumonitis in the newborn. OBJECTIVES: The objective of this review was to assess the effects of antibiotics in the treatment of genital infection with Chlamydia trachomatis during pregnancy with respect to neonatal and maternal morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (Cochrane Library issue 1, 1999). SELECTION CRITERIA: Randomised trials of any antibiotic regimen compared with placebo or no treatment or alternative antibiotic regimens in pregnant women with genital Chlamydia trachomatis infection. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were done independently by two reviewers. Study authors were contacted for additional information. MAIN RESULTS: Eleven trials were included. Trial quality was generally good. Amoxycillin appeared to be as effective as erythromycin in achieving microbiological cure (odds ratio 0.54, 95% confidence interval 0.28 to 1.02). Amoxycillin was better tolerated than erythromycin (odds ratio 0.16, 95% confidence interval 0.09 to 0.30). Clindamycin and azithromycin also appear to be effective, although the numbers of women included in trials are small. REVIEWER'S CONCLUSIONS: Amoxycillin appears to be an acceptable alternative therapy for the treatment of genital chlamydial infections in pregnancy when compared with erythromycin. Clindamycin and azithromycin may be considered if erythromycin and amoxycillin are contra-indicated or not tolerated. PMID- 10796107 TI - Chorion villus sampling versus amniocentesis for prenatal diagnosis. AB - BACKGROUND: Amniocentesis test results are usually available only after 18 weeks gestation. Chorion villus sampling (CVS) may be performed transabdominally or transvaginally, usually between 10 and 12 weeks gestation. OBJECTIVES: The objective of this review was to assess the safety and accuracy of chorion villus sampling compared to amniocentesis. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials comparing first trimester chorion villus sampling and second trimester amniocentesis. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Three studies involving over 9000 women were included. The trials were generally of good quality. Compared to amniocentesis, chorion villus sampling was associated with more sampling and technical failures, and more false positive and false negative results. Pregnancy loss was more common after chorion villus sampling (odds ratio 1.33, 95% confidence interval 1.17 to 1.52). There is a suggestion (though not statistically significant) of an increase in stillbirths and neonatal deaths following chorion villus sampling. Maternal complications were uncommon. REVIEWER'S CONCLUSIONS: The increase in miscarriages after chorion villus sampling compared to amniocentesis appear to be procedure related. Second trimester amniocentesis appears to be safer than chorion villus sampling. The benefits of earlier diagnosis with chorion villus sampling must be set against the greater risk of pregnancy loss. PMID- 10796108 TI - Continuity of caregivers for care during pregnancy and childbirth. AB - BACKGROUND: Care is often provided by multiple caregivers, many of whom work only in the antenatal clinic, labour ward or postnatal unit. However continuity of care is provided by the same caregiver or a small group from pregnancy through the postnatal period. OBJECTIVES: The objective of this review was to assess continuity of care during pregnancy and childbirth and the puerperium with usual care by multiple caregivers. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Controlled trials comparing continuity of care with usual care during pregnancy, childbirth and the postnatal period. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Study authors were contacted for additional information. MAIN RESULTS: Two studies involving 1815 women were included. Both trials compared continuity of care by midwives with non-continuity of care by a combination of physicians and midwives. The trials were of good quality. Compared to usual care, women who had continuity of care from a team of midwives were less likely to be admitted to hospital antenatally (odds ratio 0.79, 95% confidence interval 0.64 to 0.97) and more likely to attend antenatal education programs (odds ratio 0.58, 95% confidence interval 0.41 to 0.81). They were also less likely to have drugs for pain relief during labour (odds ratio 0.53, 95% confidence interval 0.44 to 0.64) and their newborns were less likely to require resuscitation (odds ratio 0.66, 95% confidence interval 0.52 to 0.83). No differences were detected in Apgar scores, low birthweight and stillbirths or neonatal deaths. While they were less likely to have an episiotomy (odds ratio 0.75, 95% confidence interval 0.60 to 0.94), women receiving continuity of care were more likely to have either a vaginal or perineal tear (odds ratio 1.28, 95% confidence interval 1.05, 1.56). They were more likely to be pleased with their antenatal, intrapartum and postnatal care. REVIEWER'S CONCLUSIONS: Studies of continuity of care show beneficial effects. It is not clear whether these are due to greater continuity of care, or to midwifery care. PMID- 10796109 TI - Continuous electronic heart rate monitoring for fetal assessment during labor. AB - BACKGROUND: Electronic fetal monitoring (EFM) has been widely adopted. There is debate about its overall effectiveness as well as the relative merits of routine application versus use for high-risk pregnancies only. OBJECTIVES: The objective of this review was to assess the effects of routine continuous electronic fetal monitoring during labour compared with intermittent auscultation. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, Medline (1966 to 1994), and reference list of relevant articles. We also contacted experts in the field. SELECTION CRITERIA: Randomised trials comparing routine continuous electronic fetal monitoring with intermittent auscultation. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer, and their accuracy was confirmed independently by a second person. A single reviewer assessed study quality based on criteria developed by others for randomised controlled trials. Data reported from similar studies were used to calculate a combined risk estimate for each of eight outcomes. MAIN RESULTS: Nine studies involving 18,561 women and their 18,695 infants were included. The trials were of variable quality. A statistically significant decrease was associated with routine continuous EFM for neonatal seizures (relative risk (RR) = 0. 51, confidence interval (CI) = 0.32,0.82). The protective effect for neonatal seizures was only evident in studies with high-quality scores. No significant differences were observed in 1-minute Apgar scores below 4, 1-minute Apgar scores below 7, rate of admissions to neonatal intensive care units, and perinatal death. An increase associated with the use of EFM was observed in the rate of cesarean delivery (RR = 1.41, CI = 1.23,1.61) and operative vaginal delivery (RR = 1.20, CI = 1.11,1.30). REVIEWER'S CONCLUSIONS: The only clinically significant benefit from the use of routine continuous EFM was in the reduction of neonatal seizures. In view of the increase in cesarean and operative vaginal deliveries, the long-term benefit of this reduction must be evaluated in the decision reached jointly by the pregnant woman and her clinician to use continuous EFM or intermittent auscultation during labor. PMID- 10796110 TI - Prophylactic corticosteroids for preterm birth. AB - BACKGROUND: Respiratory distress syndrome is a serious complication of prematurity causing significant immediate and long-term mortality and morbidity. OBJECTIVES: The objective of this review was to assess the effects of corticosteroids administered to pregnant women to accelerate fetal lung maturity prior to preterm delivery. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised and quasi randomised trials of corticosteroid drugs capable of crossing the placenta compared with placebo or no treatment in women expected to deliver preterm as a result of either spontaneous preterm labour, prelabour rupture of the membranes preterm, or elective preterm delivery. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Eighteen trials including data on over 3700 babies were included. Antenatal administration of 24 milligrams of betamethasone, of 24 milligrams of dexamethasone, or two grams of hydrocortisone to women expected to give birth preterm was associated with a significant reduction in mortality (odds ratio 0.60, 95% confidence interval 0.48 to 0.75), respiratory distress syndrome (odds ratio 0.53, 95% confidence interval 0.44 to 0.63) and intraventricular haemorrhage in preterm infants. These benefits extended to a broad range of gestational ages and were not limited by gender or race. No adverse consequences of prophylactic corticosteroids for preterm birth have been identified. REVIEWER'S CONCLUSIONS: Corticosteroids given prior to preterm birth (as a result of either preterm labour or elective preterm delivery) are effective in preventing respiratory distress syndrome and neonatal mortality. However there is not enough evidence to evaluate the use of repeated doses of corticosteroids in women who remain undelivered, but who are at continued risk of preterm birth. (This abstract has been prepared centrally.) PMID- 10796111 TI - Creams for preventing stretch marks in pregnancy. AB - BACKGROUND: Many women develop stretch marks (striae gravidarum) during pregnancy. A number of creams have been used to remove these stretch marks. OBJECTIVES: The objective of this review was to assess the effects of topical treatments to prevent the development of stretch marks. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. In addition, the Cochrane Controlled Trials Register (CENTRAL/CCTR) was searched. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials comparing active creams with placebo for the treatment of stretch marks in pregnant women. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: One study involving 100 women was included. Compared to placebo, treatment with a cream containing Centella asiatica extract, alpha tocopherol and collagen-elastin hydrolysates was associated with less women developing stretch marks (odds ratio 0.41, 95% confidence interval 0.17 to 0.99). REVIEWER'S CONCLUSIONS: One particular cream appears to help prevent the development of stretch marks in pregnancy, but only for women who had previously suffered stretch marks in pregnancy. There is no evidence of benefit for general use. PMID- 10796112 TI - Dietary regulation for 'gestational diabetes'. AB - BACKGROUND: Impaired glucose metabolism in pregnancy may be associated with adverse pregnancy outcomes. Primary dietary therapy is used in the management of diabetes, including gestational diabetes. OBJECTIVES: The objective of this review was to assess the effects of primary dietary therapy in women identified as having gestational diabetes on fetal growth and neonatal outcomes. SEARCH STRATEGY: I searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of primary dietary therapy compared with no specific treatment in pregnant women with abnormal glucose tolerance test results. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Four studies involving 612 women were included. Trials were small and of variable quality. No differences were detected between primary dietary therapy and no primary dietary therapy for birthweight greater than 4000 grams (odds ratio 0.78, 95% confidence interval 0.45 to 1.35) or caesarean deliveries (odds ratio 0.97, 95% confidence interval 0.65 to 1.44). REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of primary dietary therapy for women who show impaired glucose metabolism during pregnancy. PMID- 10796113 TI - Doppler ultrasound for fetal assessment in high risk pregnancies. AB - BACKGROUND: Abnormal waveforms from Doppler ultrasound may indicate poor fetal prognosis. It is also possible that Doppler ultrasound could encourage inappropriate early delivery. OBJECTIVES: The objective of this review was to assess the effects of Doppler ultrasound in high risk pregnancies on obstetric care and fetal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of Doppler ultrasound for the investigation of umbilical artery waveforms in high risk pregnancies compared to no Doppler ultrasound. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by both reviewers. Study authors were contacted for additional information. MAIN RESULTS: Eleven studies involving nearly 7000 women were included. The trials were generally of good quality. Compared to no Doppler ultrasound, Doppler ultrasound in high risk pregnancy (especially those complicated by hypertension or presumed impaired fetal growth) was associated with a trend to a reduction in perinatal deaths (odds ratio 0.71, 95% confidence interval 0.50 to 1.01). The use of Doppler ultrasound was also associated with fewer inductions of labour (odds ratio 0.83, 95% confidence interval 0.74 to 0.93) and fewer admissions to hospital (odds ratio 0.56, 95% 0.43 to 0.72), without reports of adverse effects. No difference was found for fetal distress in labour (odds ratio 0.81, 95% confidence interval 0.59 to 1.13) or caesarean delivery (odds ratio 0.94, 95% 0.82 to 1.06). REVIEWER'S CONCLUSIONS: The use of Doppler ultrasound in high risk pregnancies appears to improve a number of obstetric care outcomes and appears promising in helping to reducing perinatal deaths. PMID- 10796114 TI - Doxapram treatment for apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia which may be severe enough to require resuscitation including use of positive pressure ventilation. Doxapram has been used to stimulate breathing and so prevent apnea and its consequences. OBJECTIVES: In preterm infants with recurrent apnea, does treatment with doxapram lead to a clinically important reduction in apnea and use of Intermittent positive airways pressure (IPPV), without clinically important side effects? SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal trials, the Cochrane Collaboration Clinical Trials Register, MEDLINE (using text words 'doxapram', 'apnea or apnoea' and Mesh term 'infant, premature') previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, mainly in the English language. Also an expert informant's search in the Japanese language was made by Prof. Y. Ogawa in 1996. SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which doxapram was used for the treatment of apnea in preterm infants were included. DATA COLLECTION AND ANALYSIS: Each author evaluated the papers for quality and inclusion criteria. Independent data extraction was carried out. MAIN RESULTS: Only one trial, which randomized 11 infants to intravenous doxapram and 10 infants to placebo, was found. There were fewer treatment failures after 48 hours in the group of preterm infants treated with doxapram (4/11) compared with the group treated with placebo (8/10). The wide confidence intervals made this result non significant [RR 0.45 (0.20, 1.05)]. Only one infant, who was from the placebo group, was given IPPV. Of the seven responders by 48 hours in the group of 11 who received doxapram, five failed to respond between 48 hours and seven days after commencement of therapy. This gives a late failure rate of 9/11, similar to the short term failure rate in the placebo group of 8/10. It is not possible to evaluate the late responses of all those in the placebo group since they crossed over to a treatment arm. REVIEWER'S CONCLUSIONS: Although intravenous doxapram might reduce apnea within the first 48 hours of treatment, there are insufficient data to evaluate the precision of this result or to assess potential adverse effects. No longterm outcomes have been measured. Further studies are needed to determine the role of this treatment in clinical practice. PMID- 10796115 TI - Doxapram versus methylxanthine for apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia which may be severe enough to require resuscitation including use of positive pressure ventilation. Doxapram and methylxanthine drugs have been used to stimulate breathing and so prevent apnea and its consequences. OBJECTIVES: In preterm infants with recurrent apnea, how does treatment with doxapram compare with treatment with theophylline in leading to a clinically important reduction in apnea and use of mechanical ventilation, without clinically important side effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group, as outlined in the Cochrane Library, was used. SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which doxapram was compared with methylxanthine (e.g. theophylline) for the treatment of apnea, were eligible. There must have been an effort to exclude specific causes of apnea. DATA COLLECTION AND ANALYSIS: The standard methods of the Cochrane Collaboration and its Neonatal Review Group were used to select trials, assess quality and to extract and synthesize data. The methodological quality of each trial was reviewed by the second author blinded to trial authors and institution(s). Additional information was requested from authors to clarify methodology. Each author extracted the data separately, then they were compared and differences resolved. Meta-analysis was carried out with use of relative risk and risk difference. MAIN RESULTS: In these trials involving a relatively small number of preterm infants with apnea of prematurity, there is no apparent difference between the effect of intravenous treatment with doxapram or methylxanthine on the incidence of apnea within 48 hours. There were no infants reported to have been given mechanical ventilation on either treatment. No adverse effects were reported. REVIEWER'S CONCLUSIONS: Implications for practice. The overall results of these small trials suggest that intravenous doxapram and intravenous methylxanthine are not different in their effectiveness in the short term in the treatment of apnea of prematurity. Caution is warranted as the number of patients in these trials is too small to exclude an important difference between these two treatments or to exclude the possibility of less common side effects. Longer term outcome of infants treated in these trials has not been reported. Implications for research. Further studies would require a large number of infants, stratified by gestation, to clarify which infants are likely to benefit and whether there might be differences in responses or side effects with these two drugs at different ages. It would be valuable to include important clinical outcomes such as use of mechanical ventilation as well as subsequent growth and development in future studies. Responses to treatment would have to take account of co-interventions, such as nasal continuous airway pressure which is frequently used post-intubation. PMID- 10796116 TI - Early amniocentesis versus transabdominal chorion villus sampling for prenatal diagnosis. AB - BACKGROUND: A major disadvantage of amniocentesis is that test results are usually available only after 18 weeks gestation. Early amniocentesis can now be done between 9 to 14 weeks gestation. OBJECTIVES: The objective was to assess the safety and accuracy of early amniocentesis compared with chorion villus sampling. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: October 1998. SELECTION CRITERIA: Randomised trials comparing early amniocentesis with transabdominal chorion villus sampling. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and trial quality. MAIN RESULTS: Three studies were included. Sampling failure was 0.4% in the early amniocentesis group compared to 2% in the chorion villus group (relative risk 0.23, 95% confidence interval 0.08 to 0.65). Consequently, more women in the chorion villus sampling group needed a second prenatal diagnostic test (relative risk 0.43, 95% confidence interval 0.21 to 0.88). There were no statistically significant differences in the laboratory failures (relative risk 0.43, 95% confidence interval 0. 17 to 1.10) or number of women with various chromosomal abnormalities (relative risk 0.51, 95% confidence interval 0.26 to 1. 04). Combined total pregnancy loss in the early amniocentesis group was 6.2% (57/915) compared with 5% (46/917) in the chorion villus sampling group (relative risk 1.24, 95% confidence interval 0.85 to 1.81). There were more spontaneous miscarriages after early amniocentesis (4.4% versus 2.3%, relative risk 1.92, 95% confidence interval 1.14 to 3.23). There was no difference in the incidence of neonatal respiratory distress and anomalies in the newborn infants. The incidence of talipes was greater in the early amniocentesis group, although haemangiomas were more common in the chorion villus sampling group. REVIEWER'S CONCLUSIONS: Current data suggest that early amniocentesis is associated with a greater risk of spontaneous miscarriage and neonatal talipes compared to transabdominal chorion villus sampling. An increased risk of these complications needs to be weighed against fewer technical difficulties and the possibility of fewer neonatal haemangiomas. PMID- 10796117 TI - Elective versus selective caesarean section for delivery of the small baby. AB - BACKGROUND: Elective caesarean delivery for women in preterm labour might reduce the chances of fetal or neonatal death, but it might also increase the risk of maternal morbidity. OBJECTIVES: The objective of this review was to assess the effects of a policy of elective caesarean delivery versus selective caesarean delivery for women in preterm labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised trials comparing a policy of elective caesarean delivery with expectant management with recourse to caesarean section. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and trial quality. MAIN RESULTS: Five studies involving 104 women were included. All trials reported recruiting difficulties. No significant differences between elective and selective policies for caesarean delivery were found for fetal, neonatal or maternal outcomes. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of a policy for elective caesarean delivery for small babies. Randomised trials in this area are likely to continue to experience recruitment problems. However it still may be possible to investigate elective caesarean delivery in preterm babies presenting cephalically. PMID- 10796118 TI - Transcutaneous electrostimulation for suspected placental insufficiency (diagnosed by Doppler studies). AB - BACKGROUND: Transcutaneous electrostimulation is thought to be able to improve blood flow and so it has been suggested that it may help to promote fetal growth. OBJECTIVES: The objective was to assess the effects of transcutaneous electrostimulation in suspected placental insufficiency on the promotion of fetal growth. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register and the Cochrane Controlled Trials Register. Date of last search: December 1999. SELECTION CRITERIA: Acceptably controlled trials of transcutaneous electrostimulation compared with placebo or no treatment in women with suspected impaired fetal growth or placental insufficiency. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: No studies were included. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of transcutaneous electrostimulation in the management of women with suspected placental insufficiency. PMID- 10796119 TI - Energy/protein restriction for high weight-for-height or weight gain during pregnancy. AB - OBJECTIVES: To assess the effects of prescribing a low-energy diet to pregnant women who are either overweight, or who exhibited high weight gain earlier in gestation, on subsequent weight gain, pre-eclampsia, and the outcome of pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched and researchers in the field were contacted. SELECTION CRITERIA: All acceptably controlled comparisons of protein/energy restriction prescribed to pregnant women who meet one or both of the criteria listed above. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Three studies involving 266 women were involved. Energy/protein restriction leads to a significant reduction in weekly maternal weight gain and in birth weight but has no clear effect on either pregnancy induced hypertension or pre-eclampsia. Other outcomes, including fetal/infant mortality and other measures of maternal morbidity (eg Caesarean section) or long term nutritional status, have not been reported. REVIEWER'S CONCLUSIONS: Protein/energy restriction of pregnant women who are overweight or exhibit high weight gain is unlikely to be beneficial and may be harmful to the developing fetus. PMID- 10796120 TI - Episiotomy for vaginal birth. AB - BACKGROUND: Episiotomy is done to prevent severe perineal tears, but its routine use has been questioned. The relative effects of midline compared with midlateral episiotomy are unclear. OBJECTIVES: The objective of this review was to assess the effects of restrictive use of episiotomy compared with routine episiotomy during vaginal birth. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials comparing restrictive use of episiotomy with routine use of episiotomy; restrictive use of mediolateral episiotomy versus routine mediolateral episiotomy; restrictive use of midline episiotomy versus routine midline episiotomy; and use of midline episiotomy versus mediolateral episiotomy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Six studies were included. In the routine episiotomy group, 72.7% (1752/2409) of women had episiotomies, while the rate in the restrictive episiotomy group was 27.6% (673/2441). Compared with routine use, restrictive episiotomy involved less posterior perineal trauma (relative risk 0. 88, 95% confidence interval 0.84 to 0.92), less suturing (relative risk 0.74, 95% confidence interval 0.71 to 0.77) and fewer healing complications (relative risk 0.69, 95% confidence interval 0.56 to 0.85). Restrictive episiotomy was associated with more anterior perineal trauma (relative risk 1.79, 95% 1.55 to 2.07). There was no difference in severe vaginal or perineal trauma (relative risk 1.11, 95% confidence interval 0.83 to 1.50); dyspareunia (relative risk 1.02, 95% confidence interval 0.90 to 1.16); urinary incontinence (relative risk 0.98, 95% confidence interval 0.79 to 1.20) or several pain measures. Results for restrictive versus routine mediolateral versus midline episiotomy were similar to the overall comparison. REVIEWER'S CONCLUSIONS: Restrictive episiotomy policies appear to have a number of benefits compared to routine episiotomy policies. There is less posterior perineal trauma, less suturing and fewer complications, no difference for most pain measures and severe vaginal or perineal trauma, but there was an increased risk of anterior perineal trauma with restrictive episiotomy. PMID- 10796121 TI - Expedited versus conservative approaches for vaginal delivery in breech presentation. AB - BACKGROUND: In a vaginal breech birth there may be benefit from rapid delivery of the baby to prevent progressive acidosis. However, this needs to be weighed against the potential trauma of a quick delivery. OBJECTIVES: The objective of this review was to assess the effects of expedited vaginal delivery (breech delivery from umbilicus to delivery of the head within one contraction) on perinatal outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials of expedited vaginal breech delivery compared with delivery not routinely expedited in women undergoing vaginal breech delivery. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: No studies were included. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the effects of expedited vaginal breech delivery. PMID- 10796122 TI - External cephalic version for breech presentation at term. AB - BACKGROUND: Management of breech presentation is controversial, both in regard to manipulation of the position of the fetus and the method of delivery. External cephalic version may reduce the number of breech presentations and caesarean sections, but there also have been reports of increased perinatal mortality with the procedure. OBJECTIVES: The objective of this review was to assess the effects of external cephalic version at term on measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth trials register was searched. Date of last search: October 1997. SELECTION CRITERIA: Randomised trials of external cephalic version at term (with or without tocolysis) compared with no attempt at external cephalic version in women with breech presentation. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by GJH and checked by RK. MAIN RESULTS: Six studies were included. External cephalic version at term was associated with a significant reduction in non-cephalic births (relative risk 0.42, 95% confidence interval 0.35 to 0.50) and caesarean section (relative risk 0.52, 95% confidence interval 0.39 to 0.71). There was no significant effect on perinatal mortality (relative risk 0.44, 95% confidence interval 0.07 to 2.92). REVIEWER'S CONCLUSIONS: Attempting cephalic version at term appears to reduce the chance of non-cephalic births and caesarean section. There is not enough evidence to assess any risks of external cephalic version at term. PMID- 10796123 TI - External cephalic version for breech presentation before term. AB - BACKGROUND: Management of breech presentation is controversial, both in regard to manipulation of the position of the fetus and the method of delivery. External cephalic version may reduce the number of breech presentations and caesarean sections, but there also have been reports of increased perinatal mortality with the procedure. OBJECTIVES: The objective of this review was to assess the effects of external cephalic version for breech presentation before term on measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: October 1997. SELECTION CRITERIA: Randomised and quasi-randomised trials of external cephalic version before term (37 weeks gestation) compared with a control group in women with breech presentation before term. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: Three studies involving 889 women were included. There was no significant effect of external cephalic version before term on the following: non-cephalic presentation (relative risk 1.02, 95% confidence interval 0.89 to 1.17); caesarean section (relative risk 1.10, 95% confidence interval 0.78 to 1.54); low Apgar scores (relative risk 0.81, 95% confidence interval 0.44 to 1.49); perinatal mortality (relative risk 1.19, 95% confidence interval 0.46 to 3.05). REVIEWER'S CONCLUSIONS: External cephalic version before term does not appear to improve pregnancy outcomes. PMID- 10796124 TI - Uterine exteriorization versus intraperitoneal repair at caesarean section. AB - BACKGROUND: After caesarean delivery of the fetus and placenta, the uterus may be placed outside the mother to facilitate repair of the uterine incision. OBJECTIVES: The objective of this review was to assess the effects of exteriorisation of the uterus, compared to the effects of uterine closure within the abdominal cavity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi randomised trials of uterine exteriorisation for repair compared to intra abdominal repair for pregnant women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted the data. MAIN RESULTS: Two trials involving 486 women were included. Neither trial was methodologically strong. Exteriorisation made no significant difference to blood loss. Exteriorisation was associated with fewer post operative febrile days (fever more than three days, odds ratio 0.40, 95% confidence interval 0.17 to 0.94) and a non-significant trend towards fewer infections. There was also a non-significant trend towards more nausea and vomiting when exteriorisation was done under regional analgesia. REVIEWER'S CONCLUSIONS: There is not enough information to evaluate the routine use of exteriorisation of the uterus for repair of the uterine incision. PMID- 10796125 TI - Feeding schedules in hospitals for newborn infants. AB - BACKGROUND: Regular breastfeeding times have been thought to help establish routines and promote infant digestion, while frequent breastfeeding has been recommended to enhance breastfeeding and infant growth. OBJECTIVES: The objective of this review was to assess the effects of frequent breastfeeding compared with less frequent breastfeeding in the early days after birth. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing on demand or frequent breastfeeding (two or three hourly) schedules in hospital compared with four hourly restricted feeds. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Three trials involving 400 women were included. There were significant methodological limitations in some of the studies. Compared to two hourly, three hourly or on demand breastfeeding, restricted (less frequent four hourly breastfeeding) was associated with greater discontinuation of breastfeeding by four to six weeks postpartum (relative risk 1.53, 95% confidence interval 1.08 to 2.15). Restricted breastfeeding was associated with increased incidence of sore nipples (relative risk 2.12, 95% confidence interval 1.22 to 3.68), engorgement (relative risk 2.10, 95% confidence interval 1.25 to 3.21) and the need to give additional (formula) feeds (relative risk 3.14, 95% 1.24 to 8.00). REVIEWER'S CONCLUSIONS: There appear to be a number of disadvantages from restricting breastfeeding to a four hourly schedule in the first few days after birth. More frequent or on demand breastfeeding is associated with fewer complications and longer duration of breastfeeding. PMID- 10796126 TI - Interventions for treating gonorrhoea in pregnancy. AB - BACKGROUND: Neisseria gonorrhoeae can be transmitted from the mother's genital tract to the newborn during birth and can cause gonococcal opthalmia neonatorum. OBJECTIVES: The objective of this review was to assess the effects of antibiotic regimens in the treatment of genital infection with gonorrhoea during pregnancy with respect to neonatal and maternal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: October 1998. SELECTION CRITERIA: Randomised trials of one regimen of antibiotic versus another in pregnant women with culture confirmed genital gonococcal infection. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Two trials involving 329 women were included. Amoxycillin with probenicid or spectinomycin or ceftriaxone have a similar effect on microbiological cure, defined by negative gonococcal culture. REVIEWER'S CONCLUSIONS: Any of the antibiotic regimens tested in these trials appear to be effective for the treatment of gonorrhoea in pregnancy in terms of their effect on microbiological cure. For women who are allergic to penicillin, this review provides reassurance that treatment with ceftriaxone or spectinomycin appears to be at least as equally effective in producing microbiological cure. PMID- 10796127 TI - Human chorionic gonadotrophin for recurrent miscarriage. AB - BACKGROUND: There may be an association between recurrent miscarriage and abnormal hormone function in the follicular phase. Human chorionic gonadotrophin may play a role in preventing miscarriages. OBJECTIVES: The objective of this review was to assess the effects of human chorionic gonadotrophin administration during early pregnancy on the risk of miscarriage in women with a history of recurrent miscarriage. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Date of last search: 9 January 1998. SELECTION CRITERIA: Randomised trials of human chorionic gonadotrophin compared with placebo or no treatment in women who have had two or more miscarriages. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Four trials involving 180 women were included. The trials were of variable quality. Human chorionic gonadotrophin was associated with a reduced risk of miscarriage for women with a history of recurrent miscarriage (odds ratio 0.26, 95% confidence interval 0.14 to 0.52). This result should be interpreted cautiously because the apparent effect is greatly influenced by the two methodologically weaker studies. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of human chorionic gonadotrophin during pregnancy in order to prevent miscarriage in women with a history of unexplained recurrent spontaneous miscarriage. PMID- 10796128 TI - Interventions aimed at decreasing the risk of mother-to-child transmission of HIV infection. AB - BACKGROUND: At the end of 1998 over 33 million people were infected with the human immunodeficiency virus (HIV) and over one million children had been infected from their mothers. OBJECTIVES: The objective of this review was to assess what interventions may be effective in decreasing the risk of mother-to child transmission of HIV infection as well as their effect on neonatal and maternal mortality and morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials comparing any intervention aimed at decreasing the risk of mother-to-child transmission of HIV infection compared with placebo or no treatment, or any two or more interventions aimed at decreasing the risk of mother-to-child transmission of HIV infection. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were undertaken by the reviewer. MAIN RESULTS: Zidovudine Four trials comparing zidovudine with placebo involving 1585 participants were included. Compared with placebo, there was a significant reduction in the risk of mother-to-child transmission with any zidovudine (relative risk (RR) 0.54, 95% confidence interval (CI) 0.42-0.69). There is no evidence that 'long course therapy' is superior to 'short course therapy'. Nevirapine One trial compared intrapartum and postnatal nevirapine with intrapartum and postnatal zidovudine in 626 women, the majority of whom breast fed their infants. Compared with zidovudine, there was a significant reduction in the risk of mother-to-child transmission of HIV with nevirapine (RR 0.58, 95% CI 0.40-0.83). No trials are available comparing nevirapine with placebo. Caesarean section One trial comparing elective caesarean section with anticipation of vaginal delivery involving 436 participants was included. Compared with vaginal delivery, there was a significant reduction in the risk of mother-to-child transmission of HIV infection with caesarean section (RR 0.17, 95% CI 0.05-0.55). Immunoglobulin One trial comparing hyperimmune immunoglobulin plus zidovudine with non-specific immunoglobulin plus zidovudine involving 501 participants was included. The addition of hyperimmune immunoglobulin to zidovudine does not appear to have any additional effect on the risk of mother-to-child transmission (RR 0.67, 95% CI 0.29-1.55). REVIEWER'S CONCLUSIONS: Zidovudine, nevirapine and delivery by elective caesarean section appear to be very effective in decreasing the risk of mother-to-child transmission of HIV infection. PMID- 10796129 TI - Elective high frequency oscillatory ventilation versus conventional ventilation for acute pulmonary dysfunction in preterm infants. AB - BACKGROUND: Respiratory failure due to lung immaturity is a major cause of mortality in preterm infants. Although intermittent positive pressure ventilation (IPPV) saves lives, lung distortion during its use is associated with lung injury and chronic lung disease (CLD). Conventional IPPV is provided at 30-80 breaths per minute while a newer form of ventilation called high frequency oscillatory ventilation (HFOV) provides 'breaths' at 10-15 seconds. This has been shown to result in less lung injury in experimental studies. OBJECTIVES: The objective of this review is to determine whether the elective use of high frequency oscillatory ventilation (HFOV) as compared to conventional ventilation in preterm infants who are mechanically ventilated for the respiratory distress syndrome decreases the incidence of chronic lung disease (CLD) without adverse effects. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, MEDLINE, EMBASE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching by the Cochrane Collaboration, mainly in the English language. Expert informant's search in the Japanese language was made by Prof. Y. Ogawa. SELECTION CRITERIA: Randomized controlled trials comparing HFOV and CV in preterm or low birth weight infants with pulmonary dysfunction, mainly due to RDS, who are to be given IPPV. Randomization and commencement of treatment should have been as soon as possible after the start of IPPV and usually in the first 12 hours of life. DATA COLLECTION AND ANALYSIS: The methodological quality of each trial was independently reviewed by the various authors. Each author extracted data separately; they were compared and differences were resolved. The standard method of the Cochrane Neonatal Review Group was used to synthesize the data using relative risk (RR) and risk difference (RD). From 1/RD the number needed to treat (NNT) for benefits, and number needed to harm (NNH) for adverse effects, were calculated. MAIN RESULTS: Meta-analysis of the six eligible studies comparing HFOV with CV revealed that there is no difference in mortality. There are trends toward decreases in CLD in survivors at 28-30 days, 'death or CLD at 28-30 days' and CLD in survivors at 36-37 weeks postmenstrual age or discharge in the HFOV group. However, there are trends towards increases in severe (grades 3 & 4) intraventricular hemorrhage (IVH) and in periventricular leukomalacia (PVL) in the HFOV group. HFOV results in a small increase in any air leak syndrome (ALS), [summary RR 1.20 (1.03, 1.39)]. Only 2 trials have included neurodevelopmental follow up and more survivors in the HFOV group are abnormal [summary RR 1.26 (1.01, 1.58)]. In the subgroup of four trials where a high volume strategy (HVS) was used, HFOV results in more favourable pulmonary outcomes. There are significantly lower rates of CLD in survivors at 28-30 days [summary RR 0.53 (0.36, 0.76)] and of 'death or CLD at 28-30 days' [summary RR 0.56 (0.40, 0.77) with a non-significant trend towards a reduction in oxygen use at 36-37 weeks postmenstrual age or discharge [summary RR 0.74 (0.55, 1.01)]. There were no differences in the rates of IVH or PVL. Of the four trials in the subgroup using surfactant routinely, three also used the HVS. The trends in results were similar with surfactant to those for the HVS subgroup analysis. One trial suggests that HFOV may reduce the cost of in-hospital care. In the subgroup of two trials (HIFI 1989, Rettwitz-Volk 1998) not using a HVS there is no effect of HFOV on the rate of CLD; however, there is an increase in the rate of PVL [summary RR 1.64 (1.02, 2.64). REVIEWER'S CONCLUSIONS: The overall meta-analyses is dominated by the large HIFI study which did not use the HVS recommended on the basis of animal studies, and in which surfactant was not available. Studies which used HVS have shown some benefits in short term measures of CLD without an in PMID- 10796130 TI - High protein supplementation in pregnancy. AB - OBJECTIVES: To assess the effects of providing pregnant women with high-protein nutritional supplements on gestational weight gain and on the outcome of pregnancy, including fetal growth, gestational duration, and maternal and fetal/infant morbidity and mortality. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of protein/energy supplementation in which the protein content of the supplement provided >25% of its total energy content. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Two studies involving 1076 women were included. High protein supplementation was associated with a small, statistically nonsignificant increase in weekly maternal weight gain. The two available trials provide no evidence of benefit on fetal growth; indeed, the adjusted mean difference in birth weight is -58. 4 g. One trial also reported a nonsignificantly increased risk of neonatal death with high-protein supplementation. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of high protein supplementation in pregnancy. PMID- 10796131 TI - Hormonal placental function tests for fetal assessment in high risk pregnancies. AB - BACKGROUND: Biochemical tests of fetal well-being such as placental hormone levels have not always shown direct benefits for mothers and babies. OBJECTIVES: The objective of this review was to assess the effects of measuring placental hormone levels during high risk pregnancies. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and MEDLINE. We also contacted study authors. SELECTION CRITERIA: Adequately controlled trials comparing availability with no availability of hormone placental tests in high risk pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers and this was checked by a colleague. Study authors were contacted for additional information. MAIN RESULTS: Two trials involving 3355 women were included. Methodological quality of the trials was acceptable, but results for only 230 out of 2733 women (abnormal hormone levels) were available from one trial. One trial compared oestriol levels reported promptly with levels measured but not reported. The overall perinatal mortality rate was 30.5 per 1000 live births weighing 500 grams or more. No beneficial effects of oestriol measurement on fetal outcome could be identified (odds ratio of perinatal death 0.87, 95% confidence interval 0.35 to 2.18). For abnormal test results only, the reported oestriol group showed a statistically non-significant trend to less perinatal mortality (odds ratio 0.48, 95% confidence interval 0.15 to 1.60). The other trial compared human placental lactogen measurements reported promptly with measurements not reported until after pregnancy. For abnormal test results, the reported group showed significantly less perinatal mortality with four (3.4%) deaths compared to 17 (15%) in the control group (odds ratio 0.25, 95% confidence interval 0.10 to 0.61). Data from normal test results was not available for this trial. REVIEWER'S CONCLUSIONS: The measurement of human placental lactogen levels may be of some value in high risk pregnancies, but there is not enough evidence to evaluate the use of hormone placental tests generally. PMID- 10796132 TI - Hormones for suspected impaired fetal growth. AB - BACKGROUND: It has been suggested that oestrogens may improve fetal growth due to an increase in nutritional supply to the fetus from greater uterine blood flow. OBJECTIVES: The objective of this review was to assess the effects of hormone administration for suspected impaired fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: 22 December 1999. SELECTION CRITERIA: Acceptably controlled trials of hormone administration for suspected impaired fetal growth which report fetal, perinatal or maternal outcomes. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. MAIN RESULTS: No studies were included since none of the potentially relevant trials reported clinical outcomes. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the clinical use of hormone administration for suspected impaired fetal growth. PMID- 10796133 TI - Hospitalisation and bed rest for multiple pregnancy. AB - BACKGROUND: Bed rest used to be widely advised for women with a multiple pregnancy. OBJECTIVES: The objective was to assess the effect of bed rest in hospital for women with a multiple pregnancy for prevention of preterm birth and other fetal, neonatal and maternal outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and reference lists of relevant articles were searched. Date of last search: January 1999. SELECTION CRITERIA: Randomised trials which compare outcomes in women with a multiple pregnancy and their babies who were offered bed rest in hospital with women only admitted to hospital if complications occurred. DATA COLLECTION AND ANALYSIS: Assessment for inclusion and methodological quality of the trials was done by the reviewer. Data were extracted by the reviewer and double entered. All eligible trials were included in the initial analysis. Prespecified sensitivity analyses have been carried out to evaluate the effect of trial quality, the effects of hospitalisation for bed rest in women with an uncomplicated twin pregnancy, in women with a triplet pregnancy and in women with a twin pregnancy complicated by cervical effacement and dilatation prior to labour. MAIN RESULTS: Six trials were included which involved over 600 women and 1400 babies. (1) Analyses of all trials. Routine bed rest in hospital for multiple pregnancy did not reduce the risk of preterm birth, or perinatal mortality. There was a trend to a decreased number of low birth weight infants born to women in the routinely hospitalised group, which became significant when the trial using alternate allocation was excluded (odds ratio (OR) 0.79; 95% confidence interval (CI) 0.63-0.99). No differences were seen in the number of very low birth weight infants. No support for the policy was found in other neonatal outcomes. No information is available on developmental outcomes for infants in any of the trials. Women's views about the care they received were reported rarely. (2) Analyses of hospitalisation for bed rest in women with an uncomplicated twin pregnancy. The risk of preterm birth was not reduced. Indeed significantly more women delivered very preterm (<34 weeks gestation) (OR 1.84; 95% CI 1.01-3.34). No differences were seen in perinatal mortality, or in other neonatal outcomes. Women receiving hospitalisation for bed rest had a decreased risk of developing hypertension (OR 0.55; 95% CI 0.32-0.97), although this effect was no longer apparent when the trial using alternate allocation was excluded. (3) Analyses of hospitalisation for bed rest in women with a triplet pregnancy. Most of the comparisons made between the hospitalised and control groups suggest beneficial treatment effects from routine hospitalisation for bed rest. However all the differences observed between the experimental and control groups were compatible with chance variation. (4) Analyses of hospitalisation for bed rest in women with a twin pregnancy complicated by cervical effacement and dilatation prior to labour. No differences were seen in the risk of preterm birth, perinatal mortality, fetal growth or in other neonatal outcomes. REVIEWER'S CONCLUSIONS: There is currently not enough evidence to support a policy of routine hospitalisation for bed rest in multiple pregnancy. No reduction in the risk of preterm birth or perinatal death is evident, although there is a suggestion that fetal growth is improved. For women with an uncomplicated twin pregnancy the results of this review suggest that it may be harmful in that the risk of very preterm birth is increased. Until further evidence is available to the contrary, the policy cannot be recommended for routine clinical practice. PMID- 10796135 TI - Immunotherapy for recurrent miscarriage. AB - BACKGROUND: Immunologic aberrations might be the cause of recurrent early pregnancy loss in some women. OBJECTIVES: The objective of this review was to assess the effects of leukocyte immunisation or other immunologic treatments such as intravenous immune globulin (IVIG) on the live birth rate in women with previous unexplained recurrent miscarriages. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Individual patient data were obtained from the American Society for Reproductive Immunology and by MEDLINE search. Date of last search: December 1999. SELECTION CRITERIA: Randomised trials of immunotherapy in women with three or more prior miscarriages, no more than one live birth, all non-immunologic causes ruled out and no simultaneous treatment intervention. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Eighteen trials of high quality were included. The various forms of immunotherapy did not show significant differences between treatment and control groups. For paternal cell immunisation, the odds ratios was 1.05, 95% confidence interval 0.75 to 1.47. For third party donor cell immunization, the odds ratio was 1.39, 95% confidence interval 0.68 to 2.82. For trophoblast membrane infusion the odds ratio was 0.40, 95% confidence interval 0.11 to 1.45. For intravenous immune globulin (IVIG), the odds ratio was 1.14, 95% confidence interval 0. 66 to 1.95. REVIEWER'S CONCLUSIONS: Paternal cell immunization, third party donor leukocytes, trophoblast membranes, and IVIG provide no significant beneficial effect over placebo in preventing further miscarriages. PMID- 10796134 TI - Immersion in water in pregnancy, labour and birth. AB - BACKGROUND: Immersion in warm water may help with relaxation, pain relief and increased elasticity of the birth canal. On the other hand, there may be risks associated with decreased mobility and inhalation of water by the baby, as well as increased risk of maternal infection. OBJECTIVES: The objective of this review was to assess the effects of immersion in water during pregnancy, labour or birth on maternal, fetal, neonatal and caregiver outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched and study authors were contacted. SELECTION CRITERIA: Adequately controlled trials comparing the use of any kind of bath tub which allows immersion of the abdomen in water compared with non-immersion during pregnancy, labour or birth. DATA COLLECTION AND ANALYSIS: Trials were assessed for inclusion and quality by the reviewer and a colleague. MAIN RESULTS: Three trials involving 988 women were included. No statistically significant differences between immersion and no immersion were detected for use of pain relief, augmentation and duration of first stage of labour, meconium stained liquor and perineal trauma. Neonatal outcomes such as Apgar scores, umbilical arterial pH values and neonatal infection rates also showed no differences. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of immersion in water during labour. PMID- 10796136 TI - Interventions for influencing sleep patterns in exclusively breastfed infants. AB - BACKGROUND: The difference between night and day may be reinforced in young babies by offering a 'focal feed' between 10pm and midnight and gradually lengthening the intervals between night-time feeds by a variety of other activities (such as nappy changing, re-swaddling and walking with the infant). OBJECTIVES: The objective of this review was to assess the effects of using a structured training programme to teach exclusively breastfed infants to sleep through the night by eight weeks old. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Acceptably controlled trials of a structured programme accentuating the difference between night and day compared with no intervention in first time parents of singleton babies, with mothers intending to exclusively breastfeed their infants for at least eight weeks post-partum. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: One trial involving 33 couples was included. More infants in the treatment group were sleeping throughout the night by eight weeks of age than in the control group (odds ratio 0.04, 95% confidence interval 0.01 to 0.21). REVIEWER'S CONCLUSIONS: There is some evidence to show that first time parents using a structured sleeping programme can teach their babies to sleep through the night by eight weeks of age. PMID- 10796137 TI - Instruments for transcervical chorionic villus sampling for prenatal diagnosis. AB - BACKGROUND: The type of instrument used in chorion villus sampling could have a significant impact on the success rate of the procedure. An ability to manoeuvre the instrument within the uterine cavity without puncturing the gestational sac, and to see the tip of the instrument on ultrasound scanning are particularly important. OBJECTIVES: The objective of this review was to assess the effects of different instruments for transcervical chorionic villus sampling on perinatal outcome, quality and quantity of obtained tissue, technical difficulties during the procedure and maternal adverse effects. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Date of last search: October 1998. SELECTION CRITERIA: Randomised trials comparing different instruments for transcervical chorionic villus sampling. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Three trials involving 268 women were included. None of the trials mentioned method of randomisation. An adequate sample was more likely to be obtained using small forceps than aspiration cannula (odds ratio 7.29, 95% confidence interval 3. 39 to 15.67). However small forceps were more difficult to insert than an aspiration cannula (odds ratio 0.35, 95% confidence interval 0.13 to 0.96). Use of the Portex cannula was more likely to result in an inadequate sample and a difficult or painful procedure when compared with either the silver or aluminium cannula respectively. REVIEWER'S CONCLUSIONS: Although there is some evidence to support the use of small forceps for transcervical chorionic villus sampling, it is not strong enough to support change in practice for clinicians who have become familiar with aspiration cannulas. PMID- 10796138 TI - Intrapartum antibiotics for group B streptococcal colonisation. AB - BACKGROUND: Group B streptococcal infection is common in pregnant women without causing harm. However it is also a significant cause of neonatal morbidity and mortality. OBJECTIVES: The objective of this review was to assess the effects of intrapartum administration of antibiotics to women on infant colonization with group B streptococcus, early onset neonatal group B streptococcus sepsis and neonatal death from infection. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Controlled trials of pregnant women colonized with group B streptococcus comparing intrapartum antibiotic administration with no treatment, and providing data on infant colonization with group B streptococcus and/or neonatal infection. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality assessment were done by one reviewer. MAIN RESULTS: Five trials were included. Overall quality was poor, with potential selection bias in all the identified studies. Intrapartum antibiotic treatment reduced the rate of infant colonization (odds ratio 0.10, 95% confidence interval 0.07 to 0.14) and early onset neonatal infection with group B streptococcus (odds ratio 0.17, 95% confidence interval 0.07 to 0. 39). A difference in neonatal mortality was not seen (odds ratio 0. 12, 95% confidence interval 0.01 to 2.00). REVIEWER'S CONCLUSIONS: Intrapartum antibiotic treatment of women colonized with group B streptococcus appears to reduce neonatal infection. Effective strategies to detect maternal colonization with group B streptococcus and better data on maternal risk factors for neonatal group B streptococcus infection in different populations are required. PMID- 10796139 TI - Fetal electrocardiogram plus heart rate recording for fetal monitoring during labour. AB - BACKGROUND: It is thought that adding fetal electrocardiography (ECG) to cardiotocography (CTG) alone may provide better information about fetal heart activity. OBJECTIVES: The objective of this review was to assess the effects of simultaneous recording of fetal electrocardiographic waveforms plus conventional cardiotocography, with cardiotocography alone, during labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of electrocardiogram waveform analysis with or without cardiotocography during labour. DATA COLLECTION AND ANALYSIS: Trial quality was independently assessed by two reviewers. Data extraction was done by one reviewer and checked by the second reviewer. Study authors were contacted for additional information. MAIN RESULTS: One study of 2434 pregnant women was included. This trial was of sound methodological quality. The number of operative deliveries for fetal distress was reduced by half in the electrocardiographic waveform and cardiotocography group (odds ratio 0.53, 95% confidence interval 0.39 to 0.73). There was no apparent effect on the rate of operative delivery for poor progress of labour (odds ratio 1.05, 95% confidence interval 0.87 to 1.27). The use of the electrocardiographic waveform analysis monitor had no obvious effect on the condition of the newborn. REVIEWER'S CONCLUSIONS: Analysis of the fetal electrocardiographic waveform during labour may be associated with reduced obstetric intervention without jeopardising fetal outcome. PMID- 10796140 TI - Iron supplementation in pregnancy. AB - BACKGROUND: Anaemia in pregnancy is a major health problem in many developing countries where nutritional deficiency, malaria and other parasitic infections contribute to increased maternal and perinatal mortality and morbidity. OBJECTIVES: The objective of this review was to assess the effects of iron supplementation on haematological and biochemical parameters, and on pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Study authors were also contacted. SELECTION CRITERIA: Acceptably controlled trials of iron supplementation for pregnant women. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. Study authors were contacted for additional information. MAIN RESULTS: Twenty trials were included. Iron supplementation raised or maintained the serum ferritin above 10 milligrams per litre. It resulted in a substantial reduction of women with a haemoglobin level below 10 or 10.5 grams in late pregnancy. Iron supplementation, however, had no detectable effect on any substantive measures of either maternal or fetal outcome. One trial, with the largest number of participants of selective versus routine supplementation, showed an increased likelihood of caesarean section and post-partum blood transfusion, but a lower perinatal mortality rate (up to 7 days after birth). REVIEWER'S CONCLUSIONS: Iron supplementation appears to prevent low haemoglobin at birth or at six weeks post partum. There is very little information on pregnancy outcomes for either mother or baby. There are few data derived from communities where iron deficiency is common and anaemia is a serious health problem. PMID- 10796142 TI - Lateral tilt for caesarean section. AB - BACKGROUND: When pregnant women near term lie in the supine position the uterus can compress the inferior vena cava, interfering with venous return to the heart. This can result in hypotension, reduced placental perfusion and decreased fetal oxygenation. OBJECTIVES: The objective of this review was to assess the effects on the fetus or newborn of lateral tilt at caesarean section compared to the operation carried out with the mother in the supine position. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised or quasi-randomised trials of lateral tilt (10 to 15 degrees) compared to supine position during caesarean section in pregnant women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done by one reviewer. MAIN RESULTS: Three trials involving 293 women were included. The trials were all methodologically poor. There were fewer low Apgar scores when lateral tilt was used, and pH measurements and oxygen saturation appeared to be better when tilt was used. REVIEWER'S CONCLUSIONS: There is not enough evidence from these trials to evaluate use of lateral tilt during caesarean section. PMID- 10796141 TI - Isocaloric balanced protein supplementation in pregnancy. AB - OBJECTIVES: To assess the effects of providing pregnant women with isocaloric protein supplements (ie where the protein replaces an equal quantity of nonprotein energy) on gestational weight gain and on the outcome of pregnancy. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of isocaloric protein supplementation, as long as the protein content of the supplement was 'balanced', ie the protein provided <25% of its total energy content. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Three trials involving 966 women were included. The results suggest a decrease in maternal weight gain and mean birth weight and an increased risk of small-for-gestational-age (SGA) births with isocaloric protein supplementation, but no effect on mean gestational age or preterm birth. The data are insufficient to exclude potentially important effects on fetal or neonatal mortality, and maternal health outcomes have not been reported. REVIEWER'S CONCLUSIONS: Balanced protein supplementation alone (ie without energy supplementation) is unlikely to be of benefit to pregnant women or their infants. PMID- 10796143 TI - Interventions for leg cramps in pregnancy. AB - BACKGROUND: Many women experience leg cramps in the second half of pregnancy. OBJECTIVES: The objective of this review was to assess the effects of treatments for leg cramps in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of treatments for leg cramps, persisting for at least two weeks, in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Three trials involving 217 women were included. The trials were of moderate quality. Compared with placebo, calcium reduced leg cramps (odds ratio 0.29, 95% confidence interval 0.15 to 0.56). However there was significant heterogeneity between these results. One trial comparing sodium chloride with placebo showed a reduction in leg cramps (odds ratio 0.08, 95% confidence interval 0.03 to 0.24). Based on one trial, there appeared to be no difference between calcium and sodium chloride. REVIEWER'S CONCLUSIONS: Both calcium and sodium chloride appear to help reduce leg cramps in pregnancy. However the results of the sodium chloride trial may no longer be relevant because of dietary changes. PMID- 10796144 TI - Early light reduction for preventing retinopathy of prematurity in very low birth weight infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To answer the question: "Among very low birth weight infants, what is the effect of reducing early environmental light exposure on the incidence of any "Acute ROP", or "Poor ROP Outcomes"? SEARCH STRATEGY: Searches were made of the Cochrane Neonatal Group Register of Controlled Trials, Medline, EMBASE, the Cochrane Library, previous reviews including cross references, abstracts, conference and symposia proceedings, and expert informants. The search terms used were [retrolental fibroplasia or retinopathy of prematurity] and [light or light/ae or lighting or lighting/ae or light/tu or lighting/st]. SELECTION CRITERIA: Randomized or quasi-randomized controlled trials that reduced light exposure to premature infants within the first 7 days following birth were considered for this review. DATA COLLECTION AND ANALYSIS: Data on clinical outcomes including any Acute ROP and Poor ROP Outcome were excerpted by both reviewers independently and consensus reached. Data analysis was conducted according to the standards of the Neonatal Cochrane Review Group. MAIN RESULTS: Data from four recent randomized trials, and one much older quasi-randomized trial failed to show any reduction in Acute ROP, or Poor ROP Outcome with the reduction of light to premature infants' retinas. The number of infants studied to date allows 95% confidence that IF there were a true difference being missed, it would be smaller than 7 percentage points on a background of 54% of all infants under 2 kg developing ROP. REVIEWER'S CONCLUSIONS: Decreasing retinal light exposure in premature infants is very unlikely to reduce the incidence of ROP. PMID- 10796145 TI - Magnesium sulphate versus diazepam for eclampsia. AB - BACKGROUND: A number of different anticonvulsants are used to control eclamptic fits and to prevent future seizures. OBJECTIVES: The objective of this review was to assess the effects of magnesium sulphate compared with diazepam when used for the care of women with eclampsia. Magnesium sulphate is compared with phenytoin and with lytic cocktail (in preparation) in other Cochrane reviews. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register and the Cochrane Controlled Trials Register, 1999 Issue 3. SELECTION CRITERIA: Randomised trials comparing magnesium sulphate (intravenous or intramuscular administration) with diazepam for women with a clinical diagnosis of eclampsia. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was done by the two reviewers. MAIN RESULTS: Five trials involving 1236 women were included. Most of these trials were of good quality. Magnesium sulphate was associated with a substantial reduction in the recurrence of convulsions, when compared to diazepam (relative risk 0.45, 95% confidence interval 0.35 to 0.58). Maternal mortality was also reduced, although this difference was borderline for statistical significance (relative risk 0.60, 95% CI 0.36-1.00). There are no differences in any other measures of outcome, except for fewer Apgar scores <7 at five minutes (relative risk 0.72, 95% CI 0.55-0.94) and in length of stay in SCBU >7 days (relative risk 0. 66, 95% CI 0.46-0.95) associated with magnesium sulphate. REVIEWER'S CONCLUSIONS: Magnesium sulphate appears to be substantially more effective than diazepam for treatment of eclampsia. PMID- 10796146 TI - Magnesium sulphate versus phenytoin for eclampsia. AB - BACKGROUND: A number of different anticonvulsants are used to control eclamptic fits and to prevent further seizures. OBJECTIVES: The objective of this review was to assess the effects of magnesium sulphate compared with phenytoin when used for the care of women with eclampsia. Magnesium sulphate is compared with diazepam and with lytic cocktail (in preparation) in other Cochrane reviews. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register and the Cochrane Controlled Trials Register, Issue 3 1999. SELECTION CRITERIA: Randomised trials comparing magnesium sulphate (intravenous or intramuscular administration) with phenytoin for women with a clinical diagnosis of eclampsia. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was done by two reviewers. MAIN RESULTS: Four trials involving 823 women were included. The trials were of good quality. Magnesium sulphate was associated with a substantial reduction in the recurrence of convulsions, when compared to phenytoin (relative risk 0.30, 95% confidence interval 0.20 to 0.46). The trend in maternal mortality favoured magnesium sulphate, but this difference was not statistically significant (RR 0.51, 95% CI 0.25-1.06). There was also a reduction in the risk of pneumonia (RR.44, 95% CI 0.24-0. 79), ventilation (RR.66, 95% CI 0.49-0.90) and admission to an intensive care unit (RR 0.67, 95% CI 0.50-0.89) associated with the use of magnesium sulphate. For the baby, magnesium sulphate was associated with fewer admissions to SCBU (RR 0.73, 95% CI 0.58-0.91) and fewer babies who died or were in SCBU for > 7 days (RR 0.77, 95% CI 0.63-0.95). REVIEWER'S CONCLUSIONS: Magnesium sulphate appears to be substantially more effective than phenytoin for treatment of eclampsia. PMID- 10796147 TI - Manual removal of placenta at caesarean section. AB - BACKGROUND: Although manual removal of the placenta is commonly carried out, opinions differ about the best technique for delivery of the placenta at caesarean section. OBJECTIVES: The objective of this review was to assess the effects of manual removal of the placenta at caesarean section. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing manual removal of placenta to spontaneous separation and controlled cord traction for delivery in pregnant women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done by one reviewer. MAIN RESULTS: Three trials involving 224 women were included. The trials were of reasonable quality. Manual removal of the placenta was associated with a clinically important and statistically significant increase in maternal blood loss (weighted mean difference 436.35, 95% confidence interval 347.82 to 524.90). Manual removal was also associated with increased post-partum endometritis (odds ratio 5.44, 95% confidence interval 1.25 to 23.75) and a statistically non-significant trend towards an increase in feto maternal haemorrhage (odds ratio 2.19, 95% confidence interval 0.69 to 6.93). REVIEWER'S CONCLUSIONS: The evidence suggests that manual removal of the placenta at caesarean section may do more harm than good, by increasing maternal blood loss and increasing the risk of infection. PMID- 10796148 TI - Maternal antigen avoidance during lactation for preventing atopic eczema in infants. AB - OBJECTIVES: To assess the effects of prescribing an antigen avoidance diet to lactating mothers of infants with atopic eczema on the severity of the eczema. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of maternal antigen avoidance prescribed to lactating mothers of infants with atopic eczema, regardless of the degree of antigen avoidance (number of foods eliminated from the diet) or its duration. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: One trial involving 17 women was included. Based on this single small trial, maternal antigen avoidance was associated with a nonsignificant reduction in eczema severity. REVIEWER'S CONCLUSIONS: The unimpressive results of this single trial should be interpreted with caution both because of its small size (n=17) and because the trial compared exposure to cow milk and egg with exposure to soya milk (soya can itself be allergenic). Maternal reports of changes in the severity of their breast-fed infants' eczema following ingestion of certain foods should be pursued by performing multiple (preferably double-blinded) challenges and dechallenges with the suspected foods. PMID- 10796149 TI - Maternal antigen avoidance during lactation for preventing atopic disease in infants of women at high risk. AB - OBJECTIVES: To assess the effects of prescribing an antigen avoidance diet during lactation on the nutritional status of the mother and newborn and on the development of atopic disease in the child. The main focus is on women whose infants are at high risk for developing an atopic condition, based on a history of atopic disease in the mother, father, or a previous child. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of maternal antigen avoidance prescribed to lactating women at high risk, regardless of the degree of antigen avoidance (number of foods eliminated from the diet) or its duration. Trials of multimodal interventions that include manipulation of the infant's diet other than breast milk or of other nondietary aspects of the infant's environment (eg, exposure to inhaled allergens) have been excluded from the review. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Three trials involving 209 women were included. The combined data from the three available trials suggest a strong protective effect of maternal antigen avoidance on the incidence of atopic eczema during the child's first 12-18 months of life. Methodologic shortcomings in all three trials, however, argue for caution in applying these encouraging results. In particular, the high incidence of atopic eczema in the control groups of all three trials might be explained by nonblinding or de-blinding of the examining physicians. REVIEWER'S CONCLUSIONS: Prescription of an antigen avoidance diet to a high-risk woman during lactation may substantially reduce her child's risk of developing atopic eczema, but better trials are needed. PMID- 10796150 TI - Maternal antigen avoidance during pregnancy for preventing atopic disease in infants of women at high risk. AB - OBJECTIVES: To assess the effects of prescribing an antigen avoidance diet during pregnancy on the nutritional status of the mother and newborn and on the development of atopic disease in the child. The main focus is on women at high risk for giving birth to an atopic child, based on a history of atopic disease in the mother, father, or a previous child. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of maternal antigen avoidance prescribed to pregnant women at high risk, regardless of the degree of antigen avoidance (number of foods eliminated from the diet) or the time of its onset during pregnancy. Data are also included on formula-fed (ie non breastfed) infants in trials of maternal antigen avoidance intended to continue beyond pregnancy into the lactation period. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Three trials involving 504 women were included. Based on the single trial providing pertinent data, the restricted diet was associated with a small but statistically significant lower mean gestational weight gain (mean difference = 3.0% of prepregnancy weight) and a nonsignificantly higher risk of preterm birth. The combined evidence does not suggest a strong protective effect of maternal antigen avoidance on the incidence of atopic eczema or asthma during the first 12 18 months of life. Data on allergic rhinitis/conjuctivitis and urticaria are limited to a single trial each and are insufficient to draw meaningful inferences. Two trials suggest a lower incidence of skin prick tests to egg at 6 months of age, but the effect was no longer evident at 18 months, nor was any benefit apparent at either age for skin prick tests to milk. If anything, cord blood IgE levels were higher in the experimental (antigen avoidance) group. REVIEWER'S CONCLUSIONS: Prescription of an antigen avoidance diet to a high-risk woman during pregnancy is unlikely to reduce substantially her risk of giving birth to an atopic child. Moreover, such a diet may have an adverse effect on maternal and/or fetal nutrition. PMID- 10796151 TI - Maternal hydration for increasing amniotic fluid volume in oligohydramnios and normal amniotic fluid volume. AB - BACKGROUND: Oligohydramnios (reduced amniotic fluid) may be responsible for malpresentation problems, umbilical cord compression, concentration of meconium in the liquor, and difficult or failed external cephalic version. Simple maternal hydration has been suggested as a way of increasing amniotic fluid volume in order to reduce some of these problems. OBJECTIVES: The objective of this review was to assess the effects of maternal hydration on amniotic fluid volume and measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: 1 February 1999. SELECTION CRITERIA: Randomised trials comparing maternal hydration with no hydration in pregnant women with reduced or normal amniotic fluid volume. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by both reviewers. MAIN RESULTS: Two studies of 77 women were included. The women were asked to drink two litres of water before having a repeat ultrasound examination. Maternal hydration in women with and without oligohydramnios was associated with an increase in amniotic volume (weighted mean difference for women with oligohydramnios 2.01, 95% confidence interval 1.43 to 2.56; and weighted mean difference for women with normal amniotic fluid volume 4.5, 95% confidence interval 2.92 to 6.08). Intravenous hypotonic hydration in women with oligohydramnios was associated with an increase in amniotic fluid volume (weighted mean difference 2.3, 95% confidence interval 1.36 to 3.24). Isotonic intravenous hydration had no measurable effect. No clinically important outcomes were assessed in any of the trials. REVIEWER'S CONCLUSIONS: Simple maternal hydration appears to increase amniotic fluid volume and may be beneficial in the management of oligohydramnios and prevention of oligohydramnios during labour or prior to external cephalic version. Controlled trials are needed to assess the clinical benefits and possible risks of maternal hydration for specific clinical purposes. PMID- 10796152 TI - Maternal iodine supplements in areas of deficiency. AB - BACKGROUND: Iodine deficiency is the leading preventable cause of intellectual impairment in the world. Although iodine supplementation is generally considered to be safe, there is a possibility of high doses of iodine suppressing maternal thyroid function. OBJECTIVES: The objective of this review was to assess the effects of iodine supplementation before or during pregnancy in areas of iodine deficiency. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: All acceptably controlled trials of maternal iodine supplementation during pregnancy with clinical outcomes. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by two reviewers. MAIN RESULTS: Three trials involving 1551 women were included. In two trials, iodine supplementation was associated with a statistically significant reduction in deaths during infancy and early childhood (relative risk 0.71, 95% confidence interval 0. 56 to 0.90). Iodine supplementation was associated with decreased prevalence of endemic cretinism at the age of four years (relative risk 0.27, 95% confidence interval 0.12 to 0.60) and better psychomotor development scores between four to 25 months of age. REVIEWER'S CONCLUSIONS: Iodine supplementation in a population with high levels of endemic cretinism results in an important reduction in the incidence of the condition with no apparent adverse effects. PMID- 10796153 TI - Maternal oxygen administration for fetal distress. AB - BACKGROUND: Maternal oxygen administration has been used in an attempt to lessen fetal distress by increasing the available oxygen from the mother. OBJECTIVES: The objective of this review was to assess the effects of maternal oxygenation for fetal distress during labour and to assess the effects of prophylactic oxygen therapy during the second stage of labour on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: December 1997. SELECTION CRITERIA: Randomised trials comparing maternal oxygen therapy for fetal distress during labour and prophylactic oxygen therapy during the second stage of labour with a control group (dummy or no oxygen therapy). DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: No trials addressing maternal oxygen therapy for fetal distress could be located. One trial of 85 women which addressed prophylactic oxygen therapy during labour was included. Cord blood pH values of less than 7.2 were more frequently recorded in the oxygenation group than the control group (relative risk 4.83, 95% confidence interval 1.11 to 21.04). There were no other statistically significant differences between the groups. Within the oxygenation group, those receiving oxygenation for less than 10 minutes had higher umbilical artery pH values than those receiving oxygen for longer than 10 minutes. REVIEWER'S CONCLUSIONS: There is not enough evidence to support the use of prophylactic oxygen therapy for women in labour, nor to evaluate its effectiveness for fetal distress. PMID- 10796154 TI - Maternal oxygen administration for suspected impaired fetal growth. AB - BACKGROUND: Fetal hypoxaemia is often a feature of fetal growth impairment. It has been suggested that perinatal outcome after suspected impaired fetal growth might be improved by giving mothers continuous oxygen until delivery. OBJECTIVES: The objective was to assess the effects of maternal oxygen therapy in suspected impaired fetal growth on fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Acceptably controlled trials comparing maternal oxygen therapy with no oxygen therapy in suspected impaired fetal growth. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed. MAIN RESULTS: Two studies involving 62 women were included. Oxygenation compared with no oxygenation was associated with a lower perinatal mortality rate (relative risk 0.41, 95% confidence interval 0.21 to 0.78). However higher gestational age in the oxygenation groups may have accounted for the difference in mortality rates. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the benefits and risks of maternal oxygen therapy for suspected impaired fetal growth. PMID- 10796155 TI - Interventions for nausea and vomiting in early pregnancy. AB - BACKGROUND: Nausea and vomiting are the most common symptoms experienced in early pregnancy, with nausea affecting between 70 and 85% of women. About half of pregnant women experience vomiting. OBJECTIVES: The objective of this review was to assess the effects of different methods of treating nausea and vomiting in early pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials of any treatment for nausea and/or vomiting in early pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Twenty trials were included. These trials were of variable quality. Nausea treatments were different anti-histamine medications, vitamin B6 (pyridoxine), the combination tablet Debendox (Bendectin) and P6 acupressure. For hyperemesis gravidarum two trials were identified using treatments with ginger root and ACTH (adrenocorticotropic hormone) injections. Based on 13 trials, there was an overall reduction in nausea from anti-emetic medication (odds ratio 0.17, 95% confidence interval 0. 13 to 0.21). REVIEWER'S CONCLUSIONS: Anti-emetic medication appears to reduce the frequency of nausea in early pregnancy. There is some evidence of adverse effects, but there is very little information on effects on fetal outcomes. Of newer treatments, pyridoxine (vitamin B6) appears to be more effective in reducing the severity of nausea. The results from trials of P6 acupressure are equivocal. The results of the trial using adrenocorticotropic hormone to treat hyperemesis gravidarum show no evidence of benefit. Ginger may be of benefit, but the evidence so far is weak. PMID- 10796156 TI - Maternal nutrient supplementation for suspected impaired fetal growth. AB - BACKGROUND: One way of attempting to improve fetal growth has been nutrient supplementation for the mother when fetal growth is impaired. Different nutrients such as carbohydrates and amino acids have been suggested as treatments for impaired fetal growth. OBJECTIVES: The objective of this review was to assess the effects of nutrient administration for suspected fetal growth impairment on fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: December 1999. SELECTION CRITERIA: Acceptably controlled trials of nutrient administration for suspected impaired fetal growth compared to placebo or no treatment. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Three studies involving 121 women were included. They were small and/or had methodological limitations. No difference was detected in the number of small for gestational age infants whose mothers had received nutrients compared to those who did not (relative risk for calf blood extract 0.54, 95% confidence interval (CI) 0.20 to 1.47; for glucose 1.11, 95% CI 0.64 to 1.92; for galactose 0.78, 95% CI 0.39 to 1.54. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of nutrient therapy for suspected impaired fetal growth. The studies were too small to detect any rare adverse effects. PMID- 10796157 TI - Nutritional advice in pregnancy. AB - OBJECTIVES: To assess the effects of advising pregnant women to increase their energy and protein intakes on those intakes, on gestational weight gain, and on the outcome of pregnancy. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All acceptably controlled comparisons of nutritional advice, whether administered on a one-to-one basis or to groups of women. DATA COLLECTION AND ANALYSIS: Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. MAIN RESULTS: Four trials involving 1108 women were included. Advice to increase energy and protein intakes seems to be successful in achieving those goals, but the increases are lower than those reported in trials of actual protein/energy supplementation. Data concerning effects on pregnancy outcome are available only from one trial, and, given the fact that its analysis was based on individual women despite randomization by clinic, the calculated confidence intervals are undoubtedly too narrow. Moreover, the 'significant' reduction in preterm birth associated with advice is not consistent with the total absence of effect on mean gestational age. One trial found no reduction in the incidence of pre-eclampsia. No data have been reported on potential adverse effects that might accompany increased fetal size, such as an increased risk of prolonged labour or Caesarean section. REVIEWER'S CONCLUSIONS: Nutritional advice appears effective in increasing pregnant women's energy and protein intakes, but the implications for fetal, infant, or maternal health cannot be judged from the available trials. Given the rather modest health benefits demonstrated with actual protein/energy supplementation (see the Cochrane review of 'Balanced protein/energy supplementation in pregnancy'), however, the provision of such advice is unlikely to be of major importance. PMID- 10796158 TI - Oxytocin for promoting successful lactation. AB - BACKGROUND: A rise in the concentration of oxytocin causes contraction of cells around the alveoli and milk ducts, in preparation for suckling. Lactation failure may result from insufficient oxytocin. OBJECTIVES: The objective of this review was to assess the effects of using oral or nasal oxytocin on lactation. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of variable doses of oxytocin and different methods of administration versus placebo in breastfeeding women using oxytocin to augment lactation. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers. MAIN RESULTS: Four trials of 639 women were included. There was potential for significant bias in these trials. Restricted breastfeeding schedules may have contributed to inadequate production of milk by the participants. Sublingual and buccal preparations of oxytocin were associated with an increase in milk production. Oxytocin did not appear to increase the incidence of breast pain and 100 international units of oxytocin appeared to be slightly more beneficial than 10 international units. REVIEWER'S CONCLUSIONS: An appropriate dose of sublingual or buccal oxytocin may help augment lactation where necessary. However if women are encouraged and supported with unrestricted breastfeeding, the need for oxytocin would probably be diminished. PMID- 10796159 TI - Oxytocin for prelabour rupture of membranes at or near term. AB - BACKGROUND: Induction of labour after prelabour rupture of membranes may reduce the risk of neonatal infection. OBJECTIVES: The objective of this review was to assess the effects of induction of labour with oxytocin versus expectant management for prelabour rupture of membranes at or near term (34 weeks or more). SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of early use of oxytocin versus no early use of oxytocin for spontaneous rupture of membranes, before labour (34 weeks gestation or more). DATA COLLECTION AND ANALYSIS: Trials were assessed for quality and data were abstracted. MAIN RESULTS: Eighteen studies were included. The trials were of variable quality with potential for significant bias. Compared to expectant management, induction of labour by oxytocin was associated with a decreased risk of maternal infection (odds ratio for chorioamnionitis of 0.63, 95% confidence interval 0.51 to 0.78, endometritis 0.72, 95% confidence interval 0.52 to 0.99). There was also a decreased risk of neonatal infection (odds ratio 0.64, 95% confidence interval 0.44 to 0.93). The size of this effect may have been biased in favour of oxytocin. Based on one trial, women were more likely to view their care positively if labour was induced with oxytocin. Caesarean section rates were not statistically different between groups, although the trend was towards fewer interventions with expectant management. Oxytocin was associated with more frequent use of pain relief and internal fetal heart rate monitoring. Perinatal mortality rates were low and not significantly different between groups, although the trend was towards fewer deaths with induction of labour by oxytocin. REVIEWER'S CONCLUSIONS: Induction of labour by oxytocin may decrease the risk of maternal and neonatal infection compared to expectant management. Induction of labour with oxytocin does not appear to increase the rate of caesarean section, although it may increase use of pain relief and internal fetal heart rate monitoring. PMID- 10796160 TI - Prostaglandins versus oxytocin for prelabour rupture of membranes at or near term. AB - BACKGROUND: The conventional method of induction of labour is with intravenous oxytocin. More recently, induction with prostaglandins, followed by an infusion of oxytocin if necessary, has been used. OBJECTIVES: The objective of this review was to assess the effects of induction of labour with prostaglandins compared with oxytocin, at or near term. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of early stimulation of uterine contractions with prostaglandins (with or without oxytocin) versus with oxytocin alone (not combined with prostaglandins) in women with spontaneous rupture of membranes before labour (34 weeks or more gestation). DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Seventeen trials were included. Most of the trials were of moderate to good quality. Based on six trials, prostaglandins compared with oxytocin were associated with increased chorioamnionitis (odds ratio of 1.49, 95% confidence interval 1.07 to 2.09) and maternal nausea/vomiting. Based on eight trials, prostaglandins were associated with a decrease in epidural analgesia, odds ratio of 0.85, 95% confidence interval 0.73 to 0.98 and internal fetal heart rate monitoring (based on one trial). Caesarean section, endometritis and perinatal mortality were not significantly different between the groups. REVIEWER'S CONCLUSIONS: Women with prelabour rupture of membranes at or near term having their labour induced with prostaglandins appear to have a lower risk of epidural analgesia and fetal heart rate monitoring. However there appears to be an increased risk of chorioamnionitis and nausea/vomiting with prostaglandins compared to oxytocin. PMID- 10796161 TI - Prostaglandins versus oxytocin for prelabour rupture of membranes at term. AB - BACKGROUND: The conventional method of induction of labour is with intravenous oxytocin. More recently, induction with prostaglandins, followed by an infusion of oxytocin if necessary, has been used. OBJECTIVES: The objective of this review was to assess the effects of induction of labour with prostaglandins versus oxytocin for prelabour rupture of membranes at term. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of early stimulation of uterine contractions with prostaglandins (with or without oxytocin) versus with oxytocin alone (not combined with prostaglandins) in women with spontaneous rupture of membranes at term (37 weeks or more gestation). DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Eight trials were included. Based on three trials, prostaglandins compared to oxytocin were associated with increased chorioamnionitis (odds ratio of 1.51, 95% confidence interval 1.07 to 2.12) and neonatal infections (odds ratio 1.63, 95% confidence interval 1.00 to 2.66). Based on four trials, prostaglandins were associated with a decrease in epidural analgesia (odds ratio of 0.86, 95% confidence interval 0.73 to 1.00) and internal fetal heart rate monitoring (based on one trial). Caesarean section, endometritis and perinatal mortality were not significantly different between the groups. REVIEWER'S CONCLUSIONS: Women with prelabour rupture of membranes at term having their labour induced with prostaglandins appear to have a lower risk of epidural analgesia and fetal heart rate monitoring. However there appears to be an increased risk of chorioamnionitis and neonatal infections after prostaglandin induction compared to oxytocin. PMID- 10796162 TI - Pelvimetry for fetal cephalic presentations at term. AB - BACKGROUND: Pelvimetry assesses the size of a woman's pelvis by clinical examination, or by conventional X-rays, computerised tomography scanning, or magnetic resonance imaging. OBJECTIVES: The objective of this review was to assess the effects of pelvimetry (performed antenatally, intrapartum or postpartum) on the method of delivery, and on perinatal mortality and morbidity, and on maternal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Acceptably randomised comparisons of the use of pelvimetry in cephalic presentations. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by one author. MAIN RESULTS: Four trials of over 1000 women were included. The trials were generally not of good quality. Women undergoing pelvimetry were more likely to be delivered by caesarean section (odds ratio 2.17, 95% confidence interval 1.63 to 2.88). No significant impact was detected on perinatal outcome. REVIEWER'S CONCLUSIONS: There is not enough evidence to support the use of X-ray pelvimetry in women whose fetuses have a cephalic presentation. PMID- 10796163 TI - Peritoneal non-closure at caesarean section. AB - BACKGROUND: It has been suggested that the peritoneal suture might be omitted during caesarean section without adverse effects. OBJECTIVES: The objective of this review was to assess the effects of non-closure as an alternative to closure of the peritoneum at caesarean section on intra-operative and immediate postoperative outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Controlled trials comparing leaving the visceral and/or parietal peritoneum unsutured at caesarean section with a technique which involves suturing the peritoneum in women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers. MAIN RESULTS: Four trials involving 1194 women were included. Non-closure of the peritoneum saved operating time (weighted mean difference of -6.12 minutes, 95% confidence interval -8.00 to -4.27) with no significant differences in postoperative morbidity, analgesic requirements and length of hospital stay. There was a consistent, although nonsignificant, trend for improved immediate postoperative outcome if the peritoneum was not closed. REVIEWER'S CONCLUSIONS: There seems to be no significant difference in short term morbidity from non-closure of the peritoneum at caesarean section. PMID- 10796164 TI - Phenobarbital prior to preterm birth for preventing neonatal periventricular haemorrhage. AB - BACKGROUND: Preterm infants are at risk of periventricular haemorrhage. This can damage the brain and lead to neurodevelopmental abnormalities, including cerebral palsy. Phenobarbital might prevent ischaemic injury or reduce fluctuations in blood pressure and blood flow in the brain. OBJECTIVES: The objective of this review was to assess the benefits and harms of giving phenobarbital to women at risk of imminent very preterm birth with the primary aim of preventing periventricular haemorrhage in the infant. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies. Date of last search: December 1998. SELECTION CRITERIA: Randomised trials with reported data which compare outcomes, such as neonatal mortality, neonatal neurological and other morbidity, longterm neurodevelopment and maternal morbidity, following prenatal exposure to phenobarbital, with outcomes in controls with or without placebo. DATA COLLECTION AND ANALYSIS: Assessments of trial eligibility, quality and data extractions were done by the two authors independently. Eligible trials were included in the initial analysis and prespecified sensitivity analyses done to evaluate the effect of trial quality. MAIN RESULTS: Over 1600 women were entered into the eight trials included. Analyses showed a significant reduction in the rates of all grades of periventricular haemorrhage (PVH) (relative risk 0.75, 95% confidence interval 0.65 to 0.88) and severe grades PVH (3 and 4) (relative risk 0.49, 95% confidence interval 0.32 to 0.74) in infants whose mothers had been given prenatal phenobarbital. These results were influenced by the earlier trials which were of poorer quality and contributed excessive weight in the analysis due to their higher rates of severe PVH. Over time, and with improved trial quality, these beneficial effects disappeared. No difference was found in the incidence of neurodevelopmental abnormalities at paediatric follow-up assessed between 18-36 months of age. Maternal sedation was more likely in women receiving phenobarbital. REVIEWER'S CONCLUSIONS: Phenobarbital administration to women prior to preterm birth cannot be recommended for routine clinical practice. Any future trials should examine the effects of phenobarbital prior to preterm birth at gestational ages with a high risk of PVH, stratify for gestational age and ensure minimal exclusions after randomisation. Neurodevelopmental status at follow-up should be measured as the most important outcome. PMID- 10796165 TI - Planned caesarean section for term breech delivery. AB - BACKGROUND: Routine use of caesarean section for breech presentation is widespread. However poor outcomes after breech birth may be the result of underlying conditions causing breech presentation rather than damage during delivery. OBJECTIVES: The objective of this review was to assess the effects of planned caesarean section for breech presentation on measures of pregnancy outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth trials register and the Cochrane Controlled Trials register. Date of last search: October 1997. SELECTION CRITERIA: Randomised trials comparing planned caesarean section for breech presentation with planned vaginal delivery. DATA COLLECTION AND ANALYSIS: Reviewers assessed trial eligibility and quality. MAIN RESULTS: Two studies involving 313 women were included. Caesarean section was done in 93% (119/128) of women allocated to planned caesarean section and 54% (99/185) of women in the planned vaginal delivery groups. The policy of planned caesarean section was associated with significantly increased maternal morbidity (relative risk 1.31, 95% confidence interval 1.02 to 1.68) and reduced short term neonatal morbidity (relative risk 0.26, 95% confidence interval 0.08 to 0.88). The studies were unable to detect differences in brachial plexus injuries, low Apgar scores or perinatal mortality. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of a policy of planned caesarean section for breech presentation. A large Canadian trial addressing this question is currently underway. PMID- 10796166 TI - Plasma volume expansion for suspected impaired fetal growth. AB - BACKGROUND: Failure of the normal expansion of plasma volume in the mother is associated with impaired fetal growth and pre-eclampsia. OBJECTIVES: The objective of this review was to assess the effects of plasma volume expansion for suspected impaired fetal growth. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: December 1999. SELECTION CRITERIA: Randomized or quasi-randomized trials of plasma volume expansion compared to no plasma volume expansion in women with suspected impaired fetal growth. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: No studies were included. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of plasma volume expansion for suspected impaired fetal growth. PMID- 10796167 TI - Interventions for preventing or improving the outcome of delivery at or beyond term. AB - BACKGROUND: Perinatal mortality and morbidity is increased in pregnancies of more than 42 weeks that are otherwise low risk. OBJECTIVES: The objective of this review was to assess the effects of interventions aimed at either reducing the incidence or improving the outcome of post-term pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised and quasi-randomised trials of interventions involving the intention to induce labour at a specified gestational age. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. Study authors were contacted for additional information. MAIN RESULTS: Twenty-six trials of variable quality were included. There were four trials of routine early pregnancy ultrasound, two of nipple stimulation, nineteen of routine versus selective induction of labour and one of antenatal fetal monitoring. Routine early pregnancy ultrasound reduced the incidence of post-term pregnancy (odds ratio 0.68, 95% confidence interval 0.57 to 0.82). Breast and nipple stimulation at term did not affect the incidence of post-term pregnancy (odds ratio 0.52, 95% confidence interval 0.28 to 0.96). Routine induction of labour reduced perinatal mortality (odds ratio 0.20, 95% confidence interval 0.06 to 0.70). This benefit is due to the effect of induction of labour after 41 weeks. Routine induction of labour had no effect on caesarean section. REVIEWER'S CONCLUSIONS: Routine early pregnancy ultrasound examination and subsequent adjustment of delivery date appear to reduce the incidence of post-term pregnancy. Routine induction of labour after 41 weeks gestation appears to reduce perinatal mortality. There is not enough evidence to evaluate the effects of breast and nipple stimulation, or tests of fetal wellbeing. PMID- 10796168 TI - Intravenous indomethacin for preventing mortality and morbidity in very low birth weight infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: Indomethacin is used to treat symptomatic patent ductus arteriosus and may prevent or limit intraventricular haemorrhage in the neonatal period. This review examines the effectiveness of prophylactic intravenous indomethacin in reducing the mortality and morbidity associated with these conditions in infants weighing less than 1750 grams at birth. SEARCH STRATEGY: A literature search from January 1980 to October 1994 was made in three computerised data bases: Medline; Embase; and the Oxford Database of Perinatal Trials. The search was updated in February 1997. SELECTION CRITERIA: Strict selection criteria were applied to clinical trials: the population had to be newborn infants of birth weight < 1751 grams; the intervention had to be prophylactic intravenous indomethacin; the trial had to be randomised and controlled; and at least one of several prespecified outcomes had to be reported in the results. DATA COLLECTION AND ANALYSIS: The methodological quality of each study was assessed using explicit criteria. Data on relevant outcome measures were extracted on two separate occasions and, where appropriate, the results of individual trials were combined using meta-analysis techniques to provide a pooled estimate of effect. MAIN RESULTS: There is a trend towards reduced neonatal mortality in infants receiving prophylactic indomethacin, pooled relative risk (RR) = 0. 85 [95% CI 0.66 to 1.09]. The incidence of symptomatic patent ductus arteriosus is significantly reduced in treated infants, pooled RR = 0.35 [0.26 to 0.47] but there is no evidence that treatment affects respiratory outcomes. Prophylactic indomethacin significantly reduces the incidence of Grade 3 and 4 intraventricular haemorrhage in treated infants, pooled RR = 0.60 [0.43 to 0.83]. There is no evidence to suggest prophylactic indomethacin is associated with any long term adverse effect although there is a trend in treated infants towards an increased incidence of necrotizing enterocolitis, and some evidence that treatment may transiently impair renal function. There is no evidence that haemostasis is disturbed. REVIEWER'S CONCLUSIONS: Prophylactic treatment with indomethacin has a number of immediate benefits, in particular a reduction in symptomatic patent ductus arteriosus and severe intraventricular haemorrhage. There is no evidence at present of long-term harm. Further trials are needed to assess more precisely the effects, both beneficial and harmful, on short and long term outcomes. PMID- 10796169 TI - Prophylactic intravenous preloading for regional analgesia in labour. AB - BACKGROUND: Fetal heart rate changes are common following regional analgesia during labour. Reduced uterine blood flow from maternal hypotension may contribute to this. Intravenous fluid preloading (volume expansion) may help to reduce maternal hypotension. OBJECTIVES: The objective of this review was to assess the effects of prophylactic intravenous fluid preloading prior to epidural analgesia during labour on maternal and fetal well-being. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trial register and the Cochrane Controlled Trials Register were searched. Date of last search: December 1997. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing prophylactic intravenous preloading before epidural analgesia during labour with a control group (dummy or no preloading). DATA COLLECTION AND ANALYSIS: The reviewer assessed trial quality and extracted data. MAIN RESULTS: One study involving 102 women was included. There was potential for considerable bias in this trial. Preloading with intravenous fluids was associated with a reduction in hypotension (relative risk 0.07, 95% confidence interval 0.01 to 0.53). It was also associated with a reduction in fetal heart rate abnormalities (relative risk 0.36, 95% confidence interval 0.16 to 0.83). No differences were detected in other perinatal and maternal outcomes. REVIEWER'S CONCLUSIONS: There are methodological limitations in the only trial of preloading with intravenous fluids prior to epidural analgesia. However preloading may have beneficial fetal and maternal effects in healthy women. Further investigation of the risks and benefits of intravenous preloading for women with pregnancy complications is required. PMID- 10796170 TI - Prophylactic versus therapeutic amnioinfusion for oligohydramnios in labour. AB - BACKGROUND: Amnioinfusion aims to relieve umbilical cord compression during labour by infusing a liquid into the uterine cavity. OBJECTIVES: The objective of this review was to assess the effects of prophylactic amnioinfusion for oligohydramnios compared with therapeutic amnioinfusion only if fetal heart rate decelerations or thick meconium-staining of the liquor occur. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and Cochrane Controlled Trials Register were searched. Date of last search: October 1997. SELECTION CRITERIA: Randomised trials comparing prophylactic with therapeutic amnioinfusion in women with oligohydramnios but not fetal heart rate deceleration in labour. DATA COLLECTION AND ANALYSIS: The reviewer assessed trial quality and extracted data. MAIN RESULTS: Two studies of 285 women were included. No differences were found in the rate of caesarean section (relative risk 0.98, 95% confidence interval 0.58 to 1.66), or forceps delivery. There were no difference in Apgar scores, cord arterial pH, oxytocin augmentation, meconium aspiration, neonatal pneumonia or postpartum endometritis. Prophylactic amnioinfusion was associated with increased intrapartum fever (relative risk 3.48, 95% confidence interval 1.21 to 10.05). REVIEWER'S CONCLUSIONS: There appears to be no advantage of prophylactic amnioinfusion over therapeutic amnioinfusion carried out only when fetal heart rate decelerations or thick meconium-staining of the liquor occur. PMID- 10796171 TI - Prostaglandins for prelabour rupture of membranes at or near term. AB - BACKGROUND: Induction of labour after prelabour rupture of membranes may reduce the risk of neonatal infection. However an expectant approach may be less likely to result in caesarean section. OBJECTIVES: The objective of this review was to assess the effects of induction of labour with prostaglandins versus expectant management for prelabour rupture of membranes at or near term. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing early use of prostaglandins (with or without oxytocin) with no early use of prostaglandins in women with spontaneous rupture of membranes before labour, and 34 weeks or more of gestation. DATA COLLECTION AND ANALYSIS: Trials were assessed for quality and data were abstracted. MAIN RESULTS: Fifteen trials were included. Most were of moderate to good quality. Different forms of prostaglandin preparations were used in these trials and it may be inappropriate to combine their results. Induction of labour by prostaglandins was associated with a decreased risk of chorioamnionitis (odds ratio 0.77, 95% confidence interval 0.61 to 0.97) based on eight trials and admission to neonatal intensive care (odds ratio 0.79, 95% confidence interval 0. 66 to 0.94) based on seven trials. No difference was detected for rate of caesarean section, although induction by prostaglandins was associated with a more frequent maternal diarrhoea and use of anaesthesia and/or analgesia. Based on one trial, women were more likely to view their care positively if labour was induced with prostaglandins,. REVIEWER'S CONCLUSIONS: Induction of labour with prostaglandins appears to decrease the risk of maternal infection (chorioamnionitis) and admission to neonatal intensive care. Induction of labour with prostaglandins does not appear to increase the rate of caesarean section, although it is associated with more frequent maternal diarrhoea and pain relief. PMID- 10796172 TI - Pyridoxine (vitamin B6) supplementation in pregnancy. AB - BACKGROUND: Pyridoxine (vitamin B6) contributes to the development of the central nervous system and may influence brain development and cognitive function. It may also prevent dental caries and protect the placental vascular bed. OBJECTIVES: The objective of this review was to assess the effects of vitamin B6 supplementation during pregnancy and labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of pyridoxine administration compared to a control group. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers. MAIN RESULTS: One trial involving 371 women was included. Pyridoxine supplementation either as oral capsules (odds ratio 0.63, 95% confidence interval 0. 41 to 0.95) or lozenges (odds ratio 0.33, 95% confidence interval 0. 22-0.51) was associated with decreased incidence of dental decay in pregnant women. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate pyridoxine supplementation during pregnancy, although the results of one trial suggest that it may have a beneficial effect on dental decay. PMID- 10796173 TI - Regular aerobic exercise during pregnancy. AB - BACKGROUND: Physiological responses of the fetus (especially increase in heart rate) to single, brief bouts of maternal exercise have been documented frequently. OBJECTIVES: The objective of this review was to assess the effects of advising healthy pregnant women to engage in regular (at least two to three times per week) aerobic exercise on physical fitness, labour and delivery, and the outcome of pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Acceptably controlled comparisons of prescribed aerobic exercise programmes. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Five trials involving 142 women were included. The trials were not of high methodologic quality. Three trials reported significant improvement in physical fitness in the exercise group, although differences in measures used to assess fetuses prevent a quantitative pooling of results. Two small trials reported nonsignificant results on pregnancy outcomes, but apart from a reassurring absence of effect on mean gestational age [+0.3 (-0.2 to +0.9) weeks], these results allow exclusion of only extremely large effects. REVIEWER'S CONCLUSIONS: Regular aerobic exercise during pregnancy appears to improve (or maintain) physical fitness. Available data are insufficient to exclude important risks or benefits for the mother or infant. PMID- 10796174 TI - Ultrasound for fetal assessment in early pregnancy. AB - BACKGROUND: Advantages of early pregnancy ultrasound screening are thought to be more accurate calculation of gestational age, earlier identification of multiple pregnancies, and diagnosis of non-viable pregnancies and certain fetal malformations. OBJECTIVES: The objective of this review was to assess the use of routine (screening) ultrasound compared with the selective use of ultrasound in early pregnancy (ie before 24 weeks). SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (up to July 1998) were searched. SELECTION CRITERIA: Adequately controlled trials of routine ultrasound imaging in early pregnancy. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Nine trials were included. The quality of the trials was generally good. Routine ultrasound examination was associated with earlier detection of multiple pregnancies (twins undiagnosed at 26 weeks, odds ratio 0.08, 95% confidence interval 0.04 to 0.16) and reduced rates of induction of labour for post-term pregnancy (odds ratio 0. 61, 95% confidence interval 0.52 to 0.72). There were no differences detected for substantive clinical outcomes such as perinatal mortality (odds ratio 0.86, 95% confidence interval 0.67 to 1.12). Where detection of fetal abnormality was a specific aim of the examination, the number of terminations of pregnancy for fetal anomaly increased. REVIEWER'S CONCLUSIONS: Routine ultrasound in early pregnancy appears to enable better gestational age assessment, earlier detection of multiple pregnancies and earlier detection of clinically unsuspected fetal malformation at a time when termination of pregnancy is possible. However the benefits for other substantive outcomes are less clear. PMID- 10796175 TI - Folate supplementation in pregnancy. AB - BACKGROUND: Folate depletion may result in anaemia during pregnancy. OBJECTIVES: The objective of this review was to assess the effects of folate supplementation in pregnancy on haematological and biochemical parameters and measures of pregnancy outcome. This review did not address the role of periconceptual folate supplementation to diminish the risk of fetal malformation. SEARCH STRATEGY: A comprehensive electronic search included that of the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Acceptably controlled trials of folate supplementation compared with placebo or no treatment to pregnant women with normal haemoglobin levels. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Study authors were contacted for additional information when necessary. MAIN RESULTS: Twenty-one studies were included. The trials varied in quality. Compared to placebo or no supplementation, folate supplementation was associated with increased or maintained serum folate levels (odds ratio 0.18, 95% confidence interval 0.13 to 0.24) and red cell folate levels (odds ratio 0.18, 95% confidence interval 0.09 to 0.38). Folate supplementation was associated with a reduction in the proportion of women with low haemoglobin level in late pregnancy (odds ratio 0.61, 95% confidence interval 0.52 to 0.71) and megaloblastic erythropoiesis (odds ratio 0.65, 95% confidence interval 0.45 to 0.95). Apart from a possible reduction in the incidence of low birthweight, folate supplementation appears to have no measurable effect on any other substantive measures of pregnancy outcome. REVIEWER'S CONCLUSIONS: Folate supplementation during pregnancy appears to improve haemoglobin levels and folate status. There is not enough evidence to evaluate whether folate supplementation has any effect, beneficial or harmful, on clinical outcomes for mother and baby. PMID- 10796176 TI - External cephalic version facilitation for breech presentation at term. AB - BACKGROUND: Successful external cephalic version at a late stage of pregnancy was considered to be possible only with the use of tocolytic drugs to relax the uterus. Other methods are also used in an attempt to facilitate external cephalic version at term. OBJECTIVES: The objective of this review was to assess the effects of routine tocolysis, fetal acoustic stimulation, epidural anaesthesia and transabdominal amnioinfusion for external cephalic version at term on successful version and measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing routine versus selective tocolysis; fetal acoustic stimulation in midline fetal spine positions versus dummy or no stimulation; epidural analgesia versus no epidural analgesia; or transabdominal amnioinfusion versus no amnioinfusion for external cephalic version at term. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: Six trials were included. Routine tocolysis was associated with fewer failures of external cephalic version (relative risk 0.77, 95% confidence interval 0.64 to 0.92). There were no significant differences between non-cephalic presentations and caesarean sections. Fetal acoustic stimulation in midline fetal spine positions was associated with fewer failures of external cephalic version at term (relative risk 0.17, 95% confidence interval 0.05 to 0.60). No randomised trials of epidural analgesia or transabdominal amnioinfusion for external cephalic version at term were located. REVIEWER'S CONCLUSIONS: Routine tocolysis appears to reduce the failure rate of external cephalic version at term. Although promising, there is not enough evidence to evaluate the use of fetal acoustic stimulation in midline fetal spine positions. There is not enough evidence to evaluate the use of epidural analgesia or transabdominal amnioinfusion for external cephalic version at term. PMID- 10796177 TI - Single versus two layer suturing for closing the uterine incision at caesarean section. AB - BACKGROUND: A single layer closure might save operating time, disrupt less tissue, introduce less foreign material into the wound, and thus result in less post operative morbidity. OBJECTIVES: The objective of this review was to assess the effects of a single layer closure of the uterine incision at caesarean section, in contrast to conventional two layer closure. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of single layer closure, either interrupted or continuous, compared to two layer closure with continuous sutures in pregnant women undergoing elective or emergency caesarean section. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Two trials involving 1006 women were included. Based on one trial, single layer closure was associated with reduced operating time (5.6 minutes). Based on one trial, radiographic scar appearance showed fewer scar defects at three months with the single closure group. There were no statistically significant differences in the use of extra haemostatic sutures, incidence of endometritis, decrease in post operative haematocrit or use of blood transfusion. REVIEWER'S CONCLUSIONS: There appear to be no advantages or disadvantages for routine use of single layer closure compared to two layer closure, except perhaps a shorter operation time. PMID- 10796178 TI - Support during pregnancy for women at increased risk. AB - BACKGROUND: Studies consistently show a relationship between social disadvantage and low birthweight. OBJECTIVES: The objective of this review was to assess the effects of programs offering additional social support for pregnant women who are believed to be at risk for giving birth to preterm or low birthweight babies. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials of additional support during at-risk pregnancy by either a professional (social worker, midwife, or nurse) or specially trained lay person, compared to routine care. Additional support was defined as some form of emotional support (eg counselling, reassurance, sympathetic listening) and information/advice, either in home visits or during clinic appointments, and could include tangible assistance (eg transportation to clinic appointments, assistance with the care of other children at home). DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Fourteen trials, involving over 11,000 women, were included. The trials were generally of good to excellent quality, although three used a method likely to introduce bias. Programs offering additional social support for at risk pregnant women were not associated with improvements in any medical outcomes for the index pregnancy. Some improvements in immediate psychosocial outcomes were found in individual trials. REVIEWER'S CONCLUSIONS: Programs which offer additional support during pregnancy have not been shown to reduce the numbers of low birthweight or preterm babies or improve other important medical outcomes for mothers and babies. PMID- 10796179 TI - Caregiver support for women during childbirth. AB - BACKGROUND: Social support may include advice or information, tangible assistance and emotional support. OBJECTIVES: The objective of this review was to assess the effects of continuous support during labour (provided by health care workers or lay people) on mothers and babies. SEARCH STRATEGY: I searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials comparing continuous support during labour with usual care. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Study authors were contacted for additional information. MAIN RESULTS: Fourteen trials, involving more than 5000 women, are included in the Review. The continuous presence of a support person reduced the likelihood of medication for pain relief, operative vaginal delivery, caesarean delivery, and a 5-minute Apgar score less than 7. Continuous support was also associated with a slight reduction in the length of labour. Six trials evaluated the effects of support on mothers' views of their childbirth experiences; while the trials used different measures (overall satisfaction, failure to cope well during labour, finding labour to be worse than expected, and level of personal control during childbirth), in each trial the results favoured the group who had received continuous support. REVIEWER'S CONCLUSIONS: Continuous support during labour from caregivers (nurses, midwives or lay people) appears to have a number of benefits for mothers and their babies and there do not appear to be any harmful effects. PMID- 10796180 TI - Prophylactic syntometrine versus oxytocin for delivery of the placenta. AB - BACKGROUND: The routine prophylactic administration of an oxytocic agent is an integral part of active management of the third stage of labour. These agents help prevent postpartum haemorrhage. OBJECTIVES: The objective of this review was to assess the effects of ergometrine-oxytocin (syntometrine) with oxytocin alone in reducing the risk of postpartum haemorrhage (blood loss of equal to or greater than 500 millilitres) and other maternal and neonatal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Trials of oxytocic drugs (syntometrine or oxytocin) in women having the third stage of labour managed actively. DATA COLLECTION AND ANALYSIS: Eligibility, trial quality assessment and data extraction were done independently by three reviewers. Study authors were contacted for additional information. MAIN RESULTS: Six trials were included. Compared with oxytocin, ergometrine-oxytocin (syntometrine) was associated with a small reduction in the risk of postpartum haemorrhage (odds ratio 0.74, 95% confidence interval 0.65 to 0.85). This advantage was smaller but still significant when 10 international units of oxytocin was used. There was no difference seen between the groups using either five or 10 international units for blood loss equal to or greater than 1000 millilitres. Adverse effects of vomiting and hypertension were associated with the use of ergometrine-oxytocin. No significant differences were found in other maternal or neonatal outcomes. REVIEWER'S CONCLUSIONS: The use of the combination preparation syntometrine (oxytocin and ergometrine) as part of the routine active management of the third stage of labour appears to be associated with a statistically significant reduction in the risk of postpartum haemorrhage when compared to oxytocin where blood loss is less than 1000ml. No difference was seen between the groups using either five or 10 international units for blood loss equal to or greater than 1000 millilitres. This needs to be weighed against the more common adverse effects associated with the use of syntometrine. PMID- 10796181 TI - Interventions for trichomoniasis in pregnancy. AB - BACKGROUND: Vaginitis due to Trichomonas vaginalis is one of the most common of sexually transmitted diseases. Trichomoniasis affects women during pregnancy as well but it is not clearly established whether it causes preterm birth and other pregnancy complications. OBJECTIVES: The objective of this review was to assess the effects of various treatments for trichomoniasis during pregnancy. SEARCH STRATEGY: The Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: 22 December 1999. SELECTION CRITERIA: Randomised trials comparing antitrichomonas agents during pregnancy. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed by one reviewer. MAIN RESULTS: One study in which benzoylmetronidazole was compared to no treatment in low risk symptomatic and asymptomatic women was included. Of the treated women, 95% (97/102) and 93% (77/83) were free of infection on day seven and week four respectively. This equates to a relative risk of 0.11, and 95% confidence interval 0.05 to 0.24. About 75% of partners were reported to have taken the treatment. REVIEWER'S CONCLUSIONS: Metronidazole, given as a single dose, is likely to provide parasitological cure for trichomonas, but it is not known whether this treatment will have any effect on pregnancy outcomes. The cure rate could probably be higher if more partners used the treatment. PMID- 10796182 TI - Vacuum extraction versus forceps for assisted vaginal delivery. AB - BACKGROUND: Proponents of vacuum delivery argue that it should be chosen first for assisted vaginal delivery, because it is less likely to injure the mother. OBJECTIVES: The objective of this review was to assess the effects of vacuum extraction compared to forceps, on failure to achieve delivery and maternal and neonatal morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. Date of last search: February 1999. SELECTION CRITERIA: Acceptably controlled comparisons of vacuum extraction and forceps delivery. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Ten trials were included. The trials were of reasonable quality. Use of the vacuum extractor for assisted vaginal delivery when compared to forceps delivery was associated with significantly less maternal trauma (odds ratio 0.41, 95% confidence interval 0.33 to 0.50) and with less general and regional anaesthesia. There were more deliveries with vacuum extraction (odds ratio 1.69, 95% confidence interval 1.31 to 2.19). Fewer caesarean sections were carried out in the vacuum extractor group. However the vacuum extractor was associated with an increase in neonatal cephalhaematomata and retinal haemorrhages. Serious neonatal injury was uncommon with either instrument. REVIEWER'S CONCLUSIONS: Use of the vacuum extractor rather than forceps for assisted delivery appears to reduce maternal morbidity. The reduction in cephalhaematoma and retinal haemorrhages seen with forceps may be a compensatory benefit. PMID- 10796183 TI - Topical treatment for vaginal candidiasis in pregnancy. AB - BACKGROUND: Vaginal candidiasis (moniliasis or thrush) is a common and frequently distressing infection for many women. It is even more common in pregnancy. OBJECTIVES: The objective of this review was to assess the effects of different methods of treating vaginal candidiasis in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. In addition, the Cochrane Controlled Trials Register (CENTRAL/CCTR) was searched. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials of any treatment for vaginal candidiasis in pregnancy. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Twelve trials were included. Based on five trials, imidazole drugs were more effective than nystatin when treating vaginal candidiasis in pregnancy (odds ratio 0.21, 95% confidence interval 0.16 to 0.29). Nystatin,in turn was more effective than hydrargaphen in one trial.A trial of clotrimazole was more effective than placebo (odds ratio 0.14, 95% confidence interval 0.06 to 0.31). Single dose treatment was less effective than three or four days treatment when assessed by culture and by symptoms in three different trials and treatment lasting for four days was less effective than treatment for seven days (odds ratio 10.6, 95% confidence interval 4.01 to 28.05). Based on two trials, treatment for seven days was no more or less effective than treatment for 14 days (odds ratio 0.41, 95% confidence interval 0.16 to 1.05). REVIEWER'S CONCLUSIONS: Topical imidazole appears to be more effective than nystatin for treating symptomatic vaginal candidiasis in pregnancy. Treatments for seven days may be necessary. PMID- 10796184 TI - Very tight versus tight control for diabetes in pregnancy. AB - BACKGROUND: Pregnancies complicated by pre-existing insulin dependent diabetes are high risk for a number of poor pregnancy and neonatal outcomes. OBJECTIVES: The objective of this review was to assess the effects of very tight glycaemic control in established insulin dependence. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised trials comparing very tight with tight control of diabetes in pregnancy. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Two trials involving 182 women were involved. The two trials were difficult to compare. Maternal hypoglycaemia was more common among women whose diabetic control was very tight compared to tight control (odds ratio 25.96, 95% confidence interval 4.91 to 137.26) based on one trial. There was no difference detected in perinatal outcome between the groups. REVIEWER'S CONCLUSIONS: There appears to be no clear evidence of benefit from very tight glycaemic control for pregnant diabetic women. Since very strict control may have a substantial impact on lifestyle, this suggests caution in advising such a degree of control. PMID- 10796185 TI - Vitamin D supplementation in pregnancy. AB - BACKGROUND: Vitamin D deficiency can occur in people whose diet is relatively low in the vitamin and those who are not exposed to much sunlight. OBJECTIVES: The objective of this review was to assess the effects of vitamin D supplementation on pregnancy outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (October 1998). SELECTION CRITERIA: Acceptably controlled trials of vitamin D supplementation during pregnancy. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Two trials involving 232 women were included. In one trial the mothers had higher mean daily weight gain and lower number of low birthweight infants. In the other trial the supplemented group had lower birthweights. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the effects of vitamin D supplementation during pregnancy. PMID- 10796186 TI - Vitamin K prior to preterm birth for preventing neonatal periventricular haemorrhage. AB - BACKGROUND: Preterm infants are at risk of periventricular haemorrhage. This can damage the brain and lead to neurodevelopmental abnormalities, including cerebral palsy. It has been suggested that vitamin K might improve coagulation in preterm infants. OBJECTIVES: The objective of this review was to assess the effects of vitamin K administered to women at risk of imminent very preterm birth to prevent periventricular haemorrhage and associated neurological injury in the infant. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, Cochrane Controlled Trials Register, and bibliographies up to January 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials of vitamin K administered parenterally or orally to women at risk of imminent preterm birth. The primary outcomes were neonatal mortality, neonatal neurological morbidity, as measured by the presence of periventricular haemorrhage (PVH) on ultrasound during the first week of life, and long term neurodevelopment. Secondary outcomes included other neonatal morbidity and any maternal side effects. DATA COLLECTION AND ANALYSIS: Eligibility, trial quality assessment and data extraction were done independently by two reviewers. MAIN RESULTS: Five trials were included, involving more than 420 women. The trials were of variable quality. Antenatal vitamin K was associated with a non-significant trend to a reduction in all grades of periventricular haemorrhage (relative risk (RR) 0.82, 95% confidence interval (CI) 0.67-1.00) and in severe PVH (grades 3 and 4) (RR 0.75, 95% CI 0.45 1.25) for babies receiving prenatal vitamin K compared with control babies. This trend disappeared when poorer quality trials were excluded. Information on neurodevelopment was given for a small sample of children in one trial and no differences were seen. REVIEWER'S CONCLUSIONS: Vitamin K administered to women prior to very preterm birth does not appear to be able to significantly prevent periventricular haemorrhages in preterm infants. PMID- 10796187 TI - Zinc supplementation in pregnancy. AB - BACKGROUND: It has been suggested that low serum zinc levels may be associated with abnormalities of labour, although this has not yet been established. OBJECTIVES: The objective of this review was to assess the effect of zinc supplementation in pregnancy on maternal and fetal mortality and morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Acceptably controlled trials of zinc supplementation in pregnancy. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Seven trials were included. Apart from possible reduction in induction of labour, caesarean section and preterm delivery in the supplemented group, no other differences were detected between groups of women who had zinc supplementation and those who had either placebo or no zinc during pregnancy. REVIEWER'S CONCLUSIONS: There is insufficient evidence to evaluate fully the affect of zinc supplementation during pregnancy. The possible beneficial effects on preterm delivery need to be evaluated in further trials. PMID- 10796188 TI - Antibiotics for preterm labour with intact membranes. AB - BACKGROUND: There may be a link between infection and preterm birth. OBJECTIVES: Since rupture of the membranes is an important factor in the progression of preterm labour, it is important to see if antibiotics are of any benefit prior to membrane rupture. The objective of this review was to assess the effects of antibiotics administered to women in preterm labour with intact membranes, on maternal and neonatal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised trials which compared antibiotic treatment with placebo or no treatment for women in preterm labour (between 20 and 36 weeks gestation) with intact membranes. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for missing data. MAIN RESULTS: Meta analysis of the ten included trials demonstrated a statistically significant prolongation of pregnancy associated with the use of antibiotics (5. 4 days, 95% confidence interval (CI) 0.9- 9.8 days). Antibiotic treatment was also associated with a statistically significant reduction in maternal infection (odds ratio (OR) 0.59, 95% CI 0.36, 0.97) and neonatal necrotising enterocolitis (OR 0.33, 95% CI 0.13, 0.88). No statistically significant effect was detected on the neonatal outcomes of respiratory distress syndrome and neonatal sepsis, although there was a trend towards the latter (OR 0.67, 95% CI 0.42, 1.07). An increase in perinatal mortality was observed in the group receiving antibiotics (OR 3.36, 95% CI 1.21, 9.32). REVIEWER'S CONCLUSIONS: Although a prolongation in time to delivery and a trend towards a reduction in neonatal sepsis was detected, this review fails to demonstrate a clear overall benefit from antibiotic treatment for preterm labour with intact membranes on neonatal outcomes and raises concerns about increased perinatal mortality for those who received antibiotics. This treatment cannot therefore be currently recommended for routine practice. Further research is required to identify a subgroup of women (and their babies) who are more likely to experience benefit from antibiotic treatment for preterm labour prior to membrane rupture, and to identify which antibiotic or combination of antibiotics is most effective. The results of the ongoing ORACLE trial should help resolve some of the uncertainties revealed by this review. PMID- 10796189 TI - Interventions for treating bacterial vaginosis in pregnancy. AB - BACKGROUND: Bacterial vaginosis has been associated with poor perinatal outcome. Since the infections are amenable to treatment, identification during pregnancy and treatment may reduce the risk of preterm birth and its consequences. OBJECTIVES: The objective of this review was to assess the effects of antibiotic treatment of bacterial vaginosis in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials comparing one antibiotic regimen with placebo or no treatment, or which compare two or more alternative antibiotic regimens in pregnant women with bacterial vaginosis. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were done independently by three reviewers. Study authors were contacted for additional information. MAIN RESULTS: Five trials involving 1504 women were included. These trials were of good quality. Antibiotic therapy was highly effective at eradicating infection during pregnancy as judged by 'test-of-cure' following therapy (odds ratio 0.22, 95% confidence interval 0.17 to 0.27). The effect of treating bacterial vaginosis during pregnancy showed a trend to less births before 37 weeks gestation (odds ratio 0.78, 95% confidence interval 0.60 to 1.02). The prevention of preterm birth less than 37 weeks gestation was most marked in the subgroup of women with a previous preterm birth (odds ratio 0.37, 95% confidence interval 0.23 to 0. 60). REVIEWER'S CONCLUSIONS: The current evidence does not support screening and treating all pregnant women for bacterial vaginosis to prevent preterm birth and its consequences. For women with a history of a previous preterm birth there is some suggestion that detection and treatment of bacterial vaginosis early in pregnancy may prevent a proportion of these women having a further preterm birth. It is not known whether this is associated with an improvement in neonatal well-being. PMID- 10796190 TI - Caffeine versus theophylline for apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia which may be severe enough to require resuscitation including use of positive pressure ventilation. Two forms of methylxanthine (caffeine and theophylline) have been used to stimulate breathing and so prevent apnea and its consequences. OBJECTIVES: In preterm infants with recurrent apnea, does caffeine treatment compared to theophylline treatment lead to a clinically important reduction in apnea and use of mechanical ventilation without clinically important side effects? SEARCH STRATEGY: Standard strategies of the Neonatal Review Group were used. Searches were made of the Oxford Database of Perinatal Trials, MEDLINE (see main text for strategy), previous reviews including cross references, abstracts conferences and symposium proceedings, expert informants, journal handsearching mainly in the English language. Also an expert informant's search in the Japanese language was made by Prof Y. Ogawa. SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation in which caffeine was compared with theophylline for the treatment of apnea were eligible. Trial quality and eligibility were assessed independently by each author. There must have been an effort to exclude specific causes of apnea. Measures of the severity of apnea as well as the response to treatment must have been consistent with an evaluation of 'clinical apnea' as defined by the American Academy of Pediatrics (Nelson 1978). DATA COLLECTION AND ANALYSIS: Standard method of Neonatal Review Group was used. Each author assessed eligibility, trial quality and extracted data separately, then compared and resolved differences. Results are expressed as relative risk (RR) and risk difference (RD). From 1/RD the number needed to treat (NNT) was calculated. MAIN RESULTS: There is no difference in the failure rate (< 50% reduction in apnea/bradycardia) of treatment with caffeine or theophylline at 1-3 (two studies) or 5-7 days (one study). There is a higher mean rate of apnea in the standard caffeine group at 1-3 days [three studies, mean diff. 0.398 (0.334,0.463) /100min] but not at 5-7 days (two studies). Side effects, as indicated by tachycardia or feed intolerance leading to change in dosing, are lower in the caffeine group [typical RR 0.17 (0.04,0.72), RD -0.285 (-0.467, 0.104), NNT 3.5 (2.1, 9.6)]. This was consistent across the three studies. No trial reported the use of IPPV and no data are available to assess effects on growth and development. REVIEWER'S CONCLUSIONS: Caffeine appears to have similar short term effects on apnea/bradycardia to theophylline. In view of the other therapeutic advantages of caffeine (a higher therapeutic ratio, more reliable enteral absorption and a longer half life) this is the preferred treatment for apnea in preterm infants. The possibility that higher doses of caffeine might be more effective in extremely preterm infants needs further evaluation in randomized clinical trials. PMID- 10796191 TI - Carbohydrate supplementation of human milk to promote growth in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: The main objective was to determine if addition of carbohydrate supplements to human milk leads to improved growth and neurodevelopmental outcomes without significant adverse effects in preterm infants. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This includes searches of the Oxford Database of Perinatal Trials, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language. SELECTION CRITERIA: All trials utilising random or quasi-random allocation evaluating the supplementation of human milk with carbohydrate in preterm infants within a nursery setting were eligible. DATA COLLECTION AND ANALYSIS: Not applicable. MAIN RESULTS: No eligible trials were found. REVIEWER'S CONCLUSIONS: There are no studies which have specifically evaluated the addition of carbohydrate alone for the purpose of improving growth and neurodevelopmental outcomes. No recommendations for practice can be made. Research should be directed towards comparison of different quantities and types of carbohydrate in multicomponent fortifiers containing protein and minerals, specifically evaluating short-term growth and long-term growth and neurodevelopmental outcomes. PMID- 10796192 TI - Chest physiotherapy for preventing morbidity in babies being extubated from mechanical ventilation. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effects of active chest physiotherapy on babies being extubated from mechanical ventilation for neonatal respiratory failure. SEARCH STRATEGY: The standard search strategy for the Neonatal Review Group was used as outlined in the Cochrane Handbook in the Cochrane Library. SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which active chest physiotherapy was compared with non-active techniques (eg positioning and suction alone) or no intervention in the periextubation period. The methodological quality of each trial was assessed by two independent authors. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two authors. The data were analysed from 3 trials. Subgroup analysis was performed on different treatment frequencies. MAIN RESULTS: In this review of 3 small trials, 2 of which were carried out 10 & 20 years ago, no clear benefit of periextubation active chest physiotherapy can be seen. Active chest physiotherapy did not significantly reduce the rate of postextubation lobar collapse (RR 0.69;0.33,1.45), though a reduction in the use of reintubation was shown in the overall analysis (RR 0.24;0.08,0.75). Subgroup analysis of different treatment frequencies showed the same effect with more frequent treatment (1 & 2 hourly) but showed a trend to increased lobar collapse, and no reduction in the use of reintubation, with less frequent treatment (4 hourly). There is insufficient information to assess other important short and long term outcomes, including adverse effects. REVIEWER'S CONCLUSIONS: The results of this review do not allow development of clear guidelines for clinical practice. Caution is required when interpreting the possible positive effects of chest physiotherapy of a reduction in the use of reintubation and the trend for decreased post-extubation atelectasis as the numbers of babies studied are small, the results are not consistent across trials, data on safety are insufficient, and applicability to current practice may be limited. PMID- 10796193 TI - Intravenous dexamethasone for extubation of newborn infants. AB - BACKGROUND: Endotracheal tubes are foreign bodies that may injure the upper airway causing laryngeal edema. This in turn may result in failure of extubation in preterm infants. Corticosteroids have been used prophylactically to reduce upper airway obstruction and facilitate extubation. OBJECTIVES: In newborn infants having their endotracheal tube removed following a period of intermittent positive pressure ventilation (IPPV), what are the effects of intravenous corticosteroids on the incidence of endotracheal reintubation, stridor, atelectasis and adverse side effects? SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH search terms "dexamethasone", "extubat*" and "exp infant, newborn"), previous reviews including cross references, abstracts of conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language and expert informant searches in the Japanese language by Prof. Y. Ogawa. SELECTION CRITERIA: Trials were included which used random or quasi-random patient allocation, and which compared intravenous steroids given immediately prior to a planned extubation with placebo. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the two authors and analysed in Revman for all trials. Prespecified subgroup analyses were performed to examine differences in response between infants at high risk for upper airway edema and those receiving routine prophylaxis prior to extubation. MAIN RESULTS: Administration of dexamethasone prior to extubation significantly reduced the need for reintubation of the trachea. This result applies to both the high risk group and to the total population of infants enrolled. However, the incidence of extubation failure was zero in the trial that attempted to exclude infants at high risk of airway edema. The side effects of higher blood sugar levels and glycosuria were found in the 2 trials where these were sought. REVIEWER'S CONCLUSIONS: Implications for practice Dexamethasone reduces the need for endotracheal reintubation of neonates after a period of IPPV. In view of the lack of effect in low risk infants and the documented and potential side effects, it appears reasonable to restrict its use to infants at increased risk for airway edema and obstruction, such as those who have received repeated or prolonged intubations. Implications for research Issues of dosage and applicability to the extremely low birthweight population could be addressed in future trials. Longer term outcomes such as chronic lung disease, duration of assisted ventilation and length of hospital stay should also be examined. PMID- 10796194 TI - Elective high frequency jet ventilation versus conventional ventilation for respiratory distress syndrome in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: The objective of this review was to determine whether the elective (commencing soon after initiation of mechanical ventilation) use of high frequency jet ventilation (HFJV), as compared to conventional ventilation (CV) in preterm infants with respiratory distress syndrome (RDS), would decrease the incidence of chronic lung disease (CLD) without adverse effects. SEARCH STRATEGY: Randomized trials from MEDLINE were identified by means of MeSH and text words 'high frequency ventilation', 'high frequency jet ventilation', 'jet ventilation' from the years 1980 to 1997. The EMBASE database, the Oxford Database of Perinatal Trials, the Neonatal Trials Register of the Neonatal Review Group of the Cochrane Collaboration and the Cochrane library were also accessed. Proceedings of recent SPR/APS meetings were hand searched. Information was obtained from experts in the field, and cross references were checked. SELECTION CRITERIA: All randomized controlled trials of elective high frequency jet ventilation versus conventional ventilation in preterm infants born at less than 35 weeks GA or with a birth weight less than 2000 gms with respiratory distress were included in the systematic review. Trials which used HFJV to 'rescue' preterm infants due to severe respiratory distress usually beyond 24 hours, and trials that used HFJV for a mandatory time period and then switched back to CV, were not included in this review. DATA COLLECTION AND ANALYSIS: The standard methods of the Neonatal Cochrane Review Group were used, including independent trial assessment and data extraction. Data were analysed using relative risk (RR) and risk difference (RD). From 1/RD the number needed to treat (NNT) for benefits and number needed to harm (NNH) for adverse outcomes were calculated. MAIN RESULTS: Overall analysis of the three trials showed that HFJV is associated with a reduction in CLD at 36 weeks postmenstrual age in survivors [RR 0.58 (0.34, 0. 98), RD -0.138 (-0.268, -0.007), NNT 7 (4, 90)]. The use of home oxygen therapy was evaluated in only one study (Keszler 1997) and a lower rate was found in the HFJV group [RR 0.24 (0.07, 0.79), RD -0. 176 (-0.306, -0.047), NNT 5 (3, 21)]. Overall there was a trend towards an increase in the risk of PVL in the HFJV group, which was not significant. Subgroup analyses shows a significant increase in risk of PVL in the trial by Wiswell 1996 [RR 5.0 (1.19, 21.04), RD 0.250 (0.069, 0.431), NNH 4.0 (2.3,14.5)] where a 'low volume strategy' was the standard protocol for HFJV. In the other trial by Keszler 1997, where the intention was to use a 'high volume strategy', there was no significant difference in the incidence of PVL, RR 0.42 (0.14, 1.30). In the overall analysis, there were no significant differences in the incidence of neonatal mortality, IVH all grades or in grades 3 or 4 IVH. In the subgroup where 'low volume strategy' was used there was a non-significant trend toward an increase in risk of IVH all grades and grades 3 or 4 IVH. REVIEWER'S CONCLUSIONS: The overall analysis shows a benefit in pulmonary outcomes in the group electively ventilated with HFJV. Of concern is the significant increase in acute brain injury in one trial which used lower mean airway pressures when ventilating with HFJV. There are as yet no long term pulmonary or neurodevelopmental outcomes from any of the trials. Until further studies ascertain the most appropriate strategy to routinely ventilate premature infants with HFJV safely, ventilation with HFJV cannot be recommended for preterm infants with RDS. PMID- 10796195 TI - Enemas during labor. AB - BACKGROUND: The use of enemas during labor usually depends on the preference of the attending physician and available resources. However enemas cause discomfort in women and increase the costs of delivery. OBJECTIVES: The objective of this review was to assess the effects of enemas during the first stage of labor on infection rates in mothers and newborns, duration of labor, perineal wound dehiscence in the mother, perineal pain, faecal soiling and costs. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, Database of Abstracts of Reviews of Effectiveness, Medline and reference lists of articles. SELECTION CRITERIA: Randomised trials in which an enema was administered during the first stage of labor and which included assessment of possible neonatal or puerperal morbidity or mortality. DATA COLLECTION AND ANALYSIS: Selected studies were assessed by three reviewers independently. MAIN RESULTS: Two trials involving 665 women were included. These showed no clear difference in infection rates for puerperal mothers (odds ratio 0.61, 95% confidence interval 0.36 to 1.04) or newborn children. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of routine enemas during the first stage of labor. PMID- 10796196 TI - Epidural versus non-epidural analgesia for pain relief in labour. AB - BACKGROUND: Epidural analgesia is effective in reducing labour pain, but the possible adverse effects are not clear. OBJECTIVES: The objective of this review was to assess the effects of epidural analgesia on pain relief and adverse effects in labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Randomised trials comparing epidural analgesia with other forms of analgesia not involving regional blockade, or no intervention. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. Study authors were contacted for additional information. MAIN RESULTS: Eleven studies involving 3157 women were included. Epidural analgesia was associated with greater pain relief than non-epidural methods, but also with longer first and second stages of labour, an increased incidence of fetal malposition, and increased use of oxytocin and instrumental vaginal deliveries. With new trial data included, no statistically significant effect on caesarean section rates could be identified. REVIEWER'S CONCLUSIONS: Epidural analgesia appears to be very effective in reducing pain during labour, although there appear to be some potentially adverse effects. Further research is needed to investigate adverse effects and to evaluate the different techniques used in epidural analgesia. PMID- 10796197 TI - Fat supplementation of human milk for promoting growth in preterm infants. AB - BACKGROUND: For term infants, human milk provides adequate nutrition to facilitate growth, as well as potential beneficial effects on immunity and the maternal-infant emotional state. However, the role of human milk in premature infants is less well defined as it contains insufficient quantities of some nutrients to meet the estimated needs of the infant. There are potential short term and long term benefits from human milk, although observational studies have suggested that infants fed formula have a higher rate of growth than infants who are breast fed. OBJECTIVES: The main objective is to determine if addition of supplemental fat to human milk leads to improved growth and neurodevelopmental outcomes without significant adverse effects in preterm infants. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Collaborative Review Group was used. This includes searches of the Oxford Database of Perinatal Trials, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language. SELECTION CRITERIA: All trials utilizing random or quasi-random allocation to supplementation of human milk with fat or no supplementation in preterm infants within a hospital were eligible. DATA COLLECTION AND ANALYSIS: Data were extracted using the standard methods of the Cochrane Neonatal Collaborative Review Group, with separate evaluation of trial quality and data extraction by each author and synthesis of data using relative risk and weighted mean difference. MAIN RESULTS: Results are available for only one small study evaluating the effects of fat supplementation. There are insufficient data to evaluate short term or long term growth outcomes and neurodevelopmental outcomes. There are insufficient data to comment on potential adverse effects. REVIEWER'S CONCLUSIONS: There is insufficient evidence to make recommendations for practice. Further research should evaluate the practice of supplementation of human milk with fat. This may best be done in the context of the development of multicomponent fortifiers. Both short term growth outcomes and long term growth and neurodevelopmental outcomes should be evaluated. Adverse effects should be evaluated. PMID- 10796198 TI - Home versus hospital birth. AB - BACKGROUND: A meta-analysis of observational studies have suggested that planned home birth may be safe and with less interventions than planned hospital birth. OBJECTIVES: The objective of this review was to assess the effects of planned home birth compared to hospital birth on the rates of interventions, complications and morbidity as determined in randomised trials. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: September 1999. SELECTION CRITERIA: Controlled trials comparing planned hospital birth to planned home birth in selected women, assisted by an experienced home birth practitioner, and backed up by a modern hospital system in case transfer should be necessary. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by one reviewer and checked by the other reviewer. Study authors were contacted for additional information. MAIN RESULTS: One study involving 11 women was included. The trial was of reasonable quality, but was too small to be able to draw conclusions. REVIEWER'S CONCLUSIONS: There is no strong evidence to favour either planned hospital birth or planned home birth for low risk pregnant women. PMID- 10796199 TI - Intravenous immunoglobulin for preventing infection in preterm and/or low-birth weight infants. AB - BACKGROUND: Nosocomial infections continue to be a significant cause of morbidity and mortality among preterm and/or low birth weight infants. Maternal transport of immunoglobulins to the fetus mainly occurs after 32 weeks gestation and endogenous synthesis does not begin until several months after birth. Administration of intravenous immunoglobulin provides IgG that can bind to cell surface receptors, provide opsonic activity, activate complement, promote antibody dependent cytotoxicity, and improve neutrophilic chemoluminescence. Intravenous immunoglobulin thus has the potential of preventing or altering the course of nosocomial infections. OBJECTIVES: To assess the effectiveness/safety of intravenous immunoglobulin (IVIG) administration (compared to placebo or no intervention) to preterm (< 37 weeks gestational age at birth) and/or low birth weight (LBW) (< 2500 g BW) infants in preventing nosocomial infections. SEARCH STRATEGY: Medline, Embase, Cochrane Library and Reference Update Databases were searched in November 1997 using keywords: immunoglobulin and infant-newborn and random allocation or controlled trial or randomized controlled trial (RCT). The reference lists of identified RCTs, personal files and Science Citation Index were searched. No language restrictions were applied. SELECTION CRITERIA: The criteria used to select studies for inclusion in this overview were: 1) DESIGN: RCTs in which administration of IVIG was compared to a control group that received a placebo or no intervention. 2) POPULATION: preterm (< 37 weeks gestational age) and/or LBW (<2500 g) infants. 3) INTERVENTION: IVIG for the prevention of bacterial/fungal infection during initial hospital stay (8 days or longer). (Studies that were primarily designed to assess the effect of IVIG on humoral immune markers were excluded as were studies in which the follow-up period was one week or less). 4) At least one of the following outcomes was reported: sepsis, any serious infection, death from all causes, death from infection, length of hospital stay, intraventricular haemorrhage (IVH), necrotizing enterocolitis (NEC), bronchopulmonary dysplasia (BPD). DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted information for each outcome reported in each study, and one researcher (AO) checked for any discrepancies and pooled the results. Relative risk (RR) and Risk Difference (RD) with 95% confidence intervals (CI) using the fixed effects model are reported. When a statistically significant RD was found the number needed to treat (NNT) was also calculated with 95% CIs. The results include all accepted studies in which the outcome of interest was reported. When statistically significant heterogeneity was found for an outcome, secondary (sensitivity) analyses were performed including only studies of the highest quality. MAIN RESULTS: Fifteen studies met inclusion criteria. These included 5,054 preterm and/or LBW infants and reported on at least one of the outcomes of interest for this systematic review. When all studies were combined there was a statistically significant reduction in sepsis, one or more episodes [RR 0.83 (95% CI 0.72, 0.97); RD -0.028 (95% CI -0.006, -0.051); NNT 36 (95% CI 20, 167)]. There was significant between study heterogeneity. When, in a sensitivity analysis, the high quality studies were combined, the results remained significant [RR 0.78 (95% CI 0.62, 0.98); RD 0.031(95% CI -0.003, -0.059); NNT 32 (95% CI 17, 333]. For this analysis there was no statistically significant between-study heterogeneity. A statistically significant reduction was also found for any serious infection, one or more episodes, when all studies were combined [RR 0.85 (95% CI 0.75, 0. 95); RD -0.032 (95% CI -0.010, -0.054,); NNT 31 (95% CI 19, 100). There was statistically significant between-study heterogeneity. When, in a sensitivity analysis, the high quality studies were combined the results remained statistically significant [RR 0.80 (95% CI PMID- 10796200 TI - Inositol for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Inositol is an essential nutrient required by human cells in culture for growth and survival. Inositol promotes maturation of several components of surfactant and may play a critical role in fetal and early neonatal life. OBJECTIVES: To assess the effectiveness/safety of supplementary inositol in preterm infants with RDS in reducing adverse neonatal outcomes. SEARCH STRATEGY: Medline, Embase, and Reference Update Databases were searched in August 1997 using key words: inositol and infant-newborn and random allocation or controlled trial or randomized trial (RCT). The reference lists of identified RCTs, personal files and Science Citation Index were searched. Unpublished additional information was obtained from the authors of one RCT published in abstract form. SELECTION CRITERIA: All randomized controlled trials of inositol supplementation to preterm infants with a control group that received a placebo or no intervention were included. Outcomes of interest were bronchopulmonary dysplasia (BPD), death, BPD or death, retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), and sepsis. DATA COLLECTION AND ANALYSIS: Data on neonatal outcomes were abstracted independently by the two researchers and any discrepancy was resolved through consensus. Revman was used for analysis of the data. MAIN RESULTS: Four reports of three RCTs were identified. One report was a duplicate publication. The outcome of death or bronchopulmonary dysplasia was reported in two trials, and was found to be significantly reduced (RR 0.56, 95% CI 0.42, 0.77; RD -0.215, 95% CI -0.323, 0.107). The outcome of death was reported in two trials and was found to be significantly reduced (RR 0.48, 95% CI 0.28, 0.80; RD -0.131, 95% CI -0.218, 0.043). Retinopathy of prematurity, stage 4 or needing therapy, was reported in two trials, and was found to be significantly reduced (RR 0.09, 95% CI 0.01, 0.67; RD -0.078, 95% CI -0.128, -0.027). Intraventricular hemorrhage, grade III IV, was significantly decreased (RR 0.55, 95% CI 0.32, 0.95; RD -0.090, 95% CI 0.170, -0.010). Neither sepsis nor necrotizing enterocolitis outcomes were increased. When a secondary analysis was done excluding a study published in abstract form, the results differed only in that there was a significant reduction in retinopathy of prematurity, any stage (RR 0.53, 95% CI 0.29, 0.97; RD -0.082, 95% CI -0.159,-0.005). REVIEWER'S CONCLUSIONS: Inositol supplementation results in statistically significant and clinically important reductions in important short-term adverse neonatal outcomes. A multi-center RCT of appropriate size is justified to confirm these findings. PMID- 10796201 TI - Kinesthetic stimulation for preventing apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia, which may be severe enough to require resuscitation including use of positive pressure ventilation or other treatments. Physical stimulation is often used to restart breathing and it is possible that repeated stimulation, such as with an oscillating mattress (kinesthetic stimulation), might prevent apnea and its consequences. OBJECTIVES: In preterm infants at risk for apnea, does prophylactic use of kinesthetic stimulation lead to a clinically important reduction in apnea and bradycardia, and use of intemittent positive preswsure ventilation (IPPV). SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language. SELECTION CRITERIA: All trials in preterm infants at risk of developing clinical apnea which utilised random or quasi-random allocation to treatment with an oscillating mattress or control, were eligible. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used with separate evaluation of trial quality and data extraction by each author and synthesis of the data using relative risk. MAIN RESULTS: There were no differences in short term effects (apnea /bradycardia, IVH, use of IPPV, sleep/wake cycles and neurological status at discharge) or longterm effects (in one trial - growth and development to one year). REVIEWER'S CONCLUSIONS: Implications for practice. Prophylactic use of kinesthetic stimulation cannot be recommended to reduce apnea/bradycardia in preterm infants. Implications for research. There are currently no clear research questions regarding prophylactic use of kinesthetic stimulation to prevent apnea in preterm infants. PMID- 10796202 TI - Enteral antibiotics for preventing necrotising enterocolitis in low birthweight or preterm infants. AB - BACKGROUND: Necrotising enterocolitis continues to be a problem, particularly in preterm neonates. There have been reports published suggesting that the use of enteral antibiotics may be effective as prophylaxis. This systematic review was undertaken to clarify the issue. OBJECTIVES: To evaluate the benefits and harms of enteral antibiotic prophylaxis for necrotising enterocolitis in low birth weight and preterm infants. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal trials, MEDLINE (search terms: necrotizing enterocolitis, antibiotics; LIMITS: newborn infant), previous reviews with cross references, abstracts, conference and symposia proceedings, expert informants and journal handsearching in the fields of Neonatal Pediatrics and Microbiology. SELECTION CRITERIA: All randomized or quasi-randomized controlled trials where enteral antibiotics were used as prophylaxis against NEC in LBW (<2500g) and/or preterm (<37 weeks gestation) infants. DATA COLLECTION AND ANALYSIS: The standard method of the Cochrane Collaboration and its Neonatal Review Group was used. The methodological quality of each trial was reviewed by the second author who was blinded to the trial authors and institutions. Each author extracted data separately before comparison and resolution of differences. Relative risk (RR), risk difference (RD), and number needed to treat were used in the analysis. MAIN RESULTS: The administration of prophylactic enteral antibiotics resulted in a statistically significant reduction in NEC [RR 0.47 (0.23, 0.98); RD -0.072 ( 0.136, -0.008); NNT 13.9 (7.4, 125)]. There was a reduction in NEC-related deaths which was of borderline statistical significance [RR 0.16 (0.02, 1.26); RD -0.097 (-0.183, -0.010); NNT 10.3 (5.46, 100)]. There were no significant differences in all deaths (one trial only) or in NEC-like enteropathies (one trial only). There was a statistically significant increase in the incidence of colonisation with resistant bacteria [RR 1.73 (1.00, 2. 97); RD 0.123 (0.008, 0.238); NNT 8.1 (4.2, 125)]. REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the use of enteral antibiotic prophylaxis for NEC in clinical practice. To address this question further, a large trial would be required with a sample size sufficient to examine all the important benefits and harms. Adverse outcomes associated with infection, particularly with resistant bacteria, should be evaluated. PMID- 10796203 TI - Soft versus rigid vacuum extractor cups for assisted vaginal delivery. AB - BACKGROUND: The original cups used for vacuum extraction delivery of the fetus were rigid metal cups. Subsequently, soft cups of flexible materials such as silicone rubber or plastic were introduced. Soft cups are thought to have a poorer success rate than metal cups. However they are also thought to be less likely to be associated with scalp trauma and less likely to injure the mother. OBJECTIVES: The objective of this review was to assess the effects of soft versus rigid vacuum extractor cups on perineal injury, fetal scalp injury and success rate. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: February 2000. SELECTION CRITERIA: Acceptably controlled comparisons of soft versus rigid vacuum extractor cups. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Nine trials involving 1375 women were included. The trials were of average quality. Soft cups are significantly more likely to fail to achieve vaginal delivery (odds ratio 1.65, 95% confidence interval 1.19 to 2.29). However, they were associated with less scalp injury (odds ratio 0.45, 95% confidence interval 0.15 to 0.60). There was no difference between the two groups in terms of maternal injury. REVIEWER'S CONCLUSIONS: Metal cups appear to be more suitable for 'occipito-posterior', transverse and difficult 'occipito-anterior' position deliveries. The soft cups seem to be appropriate for straightforward deliveries. PMID- 10796204 TI - Stripping/sweeping the membranes for inducing labour or preventing post-term pregnancy. AB - BACKGROUND: Stripping or sweeping of the membranes aims to initiate labour through a cascade of physiological events. These may reduce pregnancy duration or pre-empt formal induction of labour with either oxytocin, prostaglandins or amniotomy. OBJECTIVES: The objective of this review was to assess the effects of membrane sweeping to promote or induce labour on maternal and perinatal outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (last searched April 1998). SELECTION CRITERIA: Adequately controlled trials of digital separation of the membranes from the lower uterine segment for inducing labour or for preventing post-term pregnancy, compared to either no vaginal examination or vaginal examination for cervical assessment only without the intention to detach the membranes. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Fourteen trials were included. Sweeping of the membranes, performed as a general policy in women at term, was associated with reduced duration of pregnancy and the frequency of pregnancy continuing beyond 41 weeks (relative risk 0.42, 95% confidence interval 0.19 to 0.93, random effects model) and 42 weeks. After sweeping of the membranes, there was a 17% risk reduction (confidence interval 6 to 29%, random effects model) in the use of more formal methods for labour induction. There was no difference in the mode of delivery or in the risk of infection. Discomfort during vaginal examination and other adverse effects (bleeding, irregular contractions) were more frequently reported by women allocated to sweeping. REVIEWER'S CONCLUSIONS: Routine use of sweeping of membranes from 38 weeks of pregnancy onwards does not seem to produce clinically important benefits. When used as a means for induction of labour, the reduction in the use of more formal methods of induction needs to be balanced against women's discomfort and other adverse effects. PMID- 10796205 TI - Support for women/families after perinatal death. AB - BACKGROUND: Provision of an empathetic caring environment, and strategies to enable the mother and family to accept the reality of perinatal death, are now part of standard nursing and social support in most of the developed world. OBJECTIVES: The objective of this review was to assess the effects of the provision of any form of medical, nursing, social or psychological support and/or counselling to mothers and families after perinatal death. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, Medline (1966 to 1998) and reference lists of articles. SELECTION CRITERIA: Randomised trials of any form of general support aimed at encouraging acceptance of loss, specific bereavement counselling, or specialised psychological support/counselling including psychotherapy for women and families experiencing perinatal death. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility of trials. MAIN RESULTS: No trials were included. REVIEWER'S CONCLUSIONS: No information is available from randomised trials to indicate whether there is or is not a benefit from providing specific psychological support or counselling after perinatal death. PMID- 10796206 TI - Antibiotics for asymptomatic bacteriuria in pregnancy. AB - BACKGROUND: Up to 30% of mothers develop acute pyelonephritis if asymptomatic bacteriuria is untreated. Asymptomatic bacteriuria may have a role in preterm birth, or it may be a marker for low socioeconomic status and thus, low birth weight. OBJECTIVES: The objective of this review was to assess the effect of antibiotic treatment for asymptomatic bacteriuria on persistent bacteriuria during pregnancy, the risk of preterm delivery and the development of pyelonephritis after delivery. SEARCH STRATEGY: I searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials comparing antibiotic treatment with placebo or no treatment in pregnant women with asymptomatic bacteriuria found on antenatal screening. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Thirteen studies were included. Overall the study quality was not strong. Antibiotic treatment compared to placebo or no treatment was effective in clearing asymptomatic bacteriuria (odds ratio 0.07, 95% confidence interval 0.05 to 0.10). The incidence of pyelonephritis was reduced (odds ratio 0.25, 95% confidence interval 0.19 to 0.32). Antibiotic treatment was also associated with a reduction in the incidence of preterm delivery or low birth weight babies (odds ratio 0.60, 95% confidence interval 0.45 to 0.80). REVIEWER'S CONCLUSIONS: Antibiotic treatment appears to be effective in reducing the risk of pyelonephritis in pregnancy. An apparent reduction in preterm delivery is consistent with current theories about the role of infection in preterm birth, but this association should be interpreted with caution. PMID- 10796207 TI - Duration of treatment for asymptomatic bacteriuria during pregnancy. AB - BACKGROUND: A Cochrane systematic review has shown that drug treatment of asymptomatic bacteriuria in pregnant women substantially decreases the risk of pyelonephritis and reduces the risk of preterm delivery. However, it is not clear whether single dose therapy is as effective as longer conventional antibiotic treatment. OBJECTIVES: The objective of this review was to assess the effects of different durations of treatment for asymptomatic bacteriuria in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and the reference lists of articles. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing antimicrobial therapeutic regimens that differed in duration (particularly comparing single dose with longer duration regimens) in pregnant women diagnosed with asymptomatic bacteriuria. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by the reviewers. MAIN RESULTS: Eight studies involving over 400 women were included. All were comparisons of single dose treatment with four to seven day treatments. The trials were generally of poor quality. No difference in 'no-cure' rate was detected between single dose and short course (4-7 day) treatment for asymptomatic bacteriuria in pregnant women (relative risk 1.13, 95% confidence interval 0.82 to 1.54) as well as in the recurrent asymptomtic bacteriuria (relative risk 1.08, 95% confidence interval 0.70 to 1.66). However these results showed significant heterogeneity. No differences were detected for preterm births and pyelonephritis although sample size of trials was small. Longer duration treatment was associated with an increase in reports of adverse effects (relative risk 0.53, 95% confidence interval 0.31 to 0.91). REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate whether single dose or longer duration doses are more effective in treating asymptomatic bacteriuria in pregnant women. Because single dose has lower cost and increases compliance, this comparison should be explored in a properly sized randomized controlled trial. PMID- 10796208 TI - Antiplatelet agents for preventing and treating pre-eclampsia. AB - BACKGROUND: Pre-eclampsia is associated with deficient intravascular production of prostacyclin, a vasodilator, and excessive production of thromboxane, a platelet-derived vasoconstrictor and stimulant of platelet aggregation. These observations led to the hypotheses that antiplatelet agents, and low dose aspirin in particular, might prevent or delay the development of pre-eclampsia. OBJECTIVES: To assess the effectiveness and safety of antiplatelet agents when given to women at risk of developing pre-eclampsia, and to those with established pre-eclampsia. SEARCH STRATEGY: This review drew on the search strategy developed for the Pregnancy and Childbirth Group as a whole. The Cochrane Controlled Trials Register was also searched, The Cochrane Library 1999 Issue 1, Embase was searched from 1994-1999 and hand searches were performed of the congress proceedings of the International and European Societies for the Study of Hypertension in Pregnancy. SELECTION CRITERIA: All randomised trials comparing antiplatelet agents with either placebo or no antiplatelet agent during pregnancy. Quasi random study designs were excluded. Participants were pregnant women considered to be at risk of developing pre-eclampsia, and those with pre eclampsia before delivery. Women treated postpartum were excluded. Interventions were any comparisons of an antiplatelet agent (such as low dose aspirin or dipyridamole) with either placebo or no antiplatelet agent. DATA COLLECTION AND ANALYSIS: Assessment of trials for inclusion in the review and extraction of data was performed independently and unblinded by two reviewers. Data were entered into the Review Manager software and double checked. MAIN RESULTS: Forty two trials involving over 32,000 women were included in this review, with 30,563 women in the prevention trials. There is a 15% reduction in the risk of pre eclampsia associated with the use of antiplatelet agents [32 trials with 29,331 women; relative risk (RR) 0.85, 95% confidence interval (0.78, 0.92); Number needed to treat (NNT) 89, (59, 167)]. This reduction is regardless of risk status at trial entry or whether a placebo was used, and irrespective of the dose of asprin or gestation at randomisation. Twenty three trials (28,268 women) reported preterm delivery. There is a small (8%) reduction in the risk of delivery before 37 completed weeks [RR 0.92, (0.88, 0.97); NNT 72 (44, 200)]. Baby deaths were reported in 30 trials (30,093 women). Overall there is a 14% reduction in baby deaths in the antiplatelet group [RR 0.86, (0. 75, 0.98); NNT 250 (125, >10000)]. Small for gestational age babies were reported in 25 trials (20,349 women), with no overall difference between the groups, RR 0.92, (0.84, 1.01). There were no significant differences between treatment and control groups in any other measures of outcome. Five trials compared antiplatelet agents with placebo or no antiplatelet agent for the treatment of pre-eclampsia. There are insufficient data for any firm conclusions about the possible effects of these agents when used for treatment of pre-eclampsia. REVIEWER'S CONCLUSIONS: Antiplatelet agents, in this review largely low dose aspirin, have small-moderate benefits when used for prevention of pre-eclampsia. Further information is required to assess which women are most likely to benefit, when treatment should be started, and at what dose. PMID- 10796209 TI - Prostaglandins for prevention of postpartum haemorrhage. AB - BACKGROUND: Prostaglandins have mainly been used for postpartum haemorrhage when other measures fail. Misoprostol, a newer and cheaper prostaglandin E1 analogue, has sometimes been used for management of the third stage of labour. OBJECTIVES: The objective of this review was to assess the effects of prophylactic prostaglandin use in the third stage of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Researchers in the field were also contacted. Date of the latest search: December 1999. SELECTION CRITERIA: Randomized or quasi-randomized trials comparing a prostaglandin agent with another uterotonic or no prophylactic uterotonic (nothing or placebo) as part of management of the third stage of labour. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed by one reviewer. MAIN RESULTS: Seven trials were included. There were six trials of injectable prostaglandins and one of rectal misoprostol. The main limitations of these studies are their small sample sizes and lack of blind assessment of the outcomes. Injectable prostaglandins were associated with decreased blood loss (weighted mean difference -72 millilitres, 95% confidence interval -70 to -75) and shortened duration of third stage when compared to other uterotonics. Severe postpartum haemorrhage (1000 millilitres or more blood loss) occurred in two of four studies and was seen in fewer women receiving prostaglandins, though the difference was not statistically significant. Adverse effects (vomiting, diarrhoea and abdominal pain) were more common with prostaglandins when compared to other uterotonic agents. REVIEWER'S CONCLUSIONS: Although injectable prostaglandins appear to be effective in preventing postpartum haemorrhage, concerns about safety and costs limit their suitability for routine prophylactic management of third stage of labour. However, injectable prostaglandins should continue to be used for the treatment of postpartum haemorrhage when other measures fail. Misoprostol is cheap, stable and seems to be safe. Trials addressing the effectiveness of misoprostol are continuing. PMID- 10796210 TI - Therapeutic ultrasound for postpartum perineal pain and dyspareunia. AB - BACKGROUND: Proponents of therapeutic ultrasound suggest it can decrease pain by resolution of inflammation processes and reducing the pressure on pain sensitive structures by haematoma and oedema. OBJECTIVES: The objective of this review was to assess the effects of therapeutic ultrasound for treating acute perineal pain, persistent perineal pain and/or dyspareunia, following childbirth. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, Embase, Cinahl (to May 1996), Physiotherapy Index (1985 to May 1996) and World Congress of Physical Therapy 1994 Conference Proceedings. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing active therapeutic ultrasound with no treatment, placebo ultrasound, or any other 'standard' or active treatment for women with acute or persistent perineal pain and/or dyspareunia following childbirth. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Four trials involving 659 women were included. The trials were of variable quality. Based on two placebo controlled trials, women treated with active ultrasound for acute perineal pain were more likely to report improvement in pain with treatment (odds ratio 0.37, 95% confidence interval 0.19 to 0.69). No other outcome reached significance. In one trial comparing pulsed electromagnetic energy with ultrasound for acute perineal pain, women treated with ultrasound were more likely to have bruising at 10 days (odds ratio 1.64, 95% confidence interval 1.04 to 2.60). However those treated with ultrasound were less likely to have experienced perineal pain at 10 days (odds ratio 0.56, 95% confidence interval 0.34 to 0.92) and three months (odds ratio 0.43, 95% confidence interval 0.22 to 0.84). No other outcome reached significance. Based on one trial, women treated with ultrasound for persistent perineal pain and/or dyspareunia were less likely to report pain with sexual intercourse compared with the placebo group (odds ratio 0.31, 95% confidence interval 0.11 to 0.84). None of the other outcomes measured reached significance. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of ultrasound in treating perineal pain and/or dyspareunia following childbirth. PMID- 10796211 TI - Intraventricular streptokinase after intraventricular hemorrhage in newborn infants. AB - BACKGROUND: Hydrocephalus following intraventricular hemorrhage (IVH) is still one of the most serious complications of premature birth. Ventriculoperitoneal shunt surgery cannot be carried out early and permanent dependence on a shunt is associated with several serious complications. OBJECTIVES: To determine whether intraventricular streptokinase after intraventricular hemorrhage reduces the risk of permanent shunt dependence, neurodevelopmental disability or death in neonates at risk of, or actually developing, post-hemorrhagic hydrocephalus (PHH). This form of therapy is based on the hypothesis that multiple blood clots in the CSF are the initial cause of post-hemorrhagic ventricular dilatation and lysis of clots could reopen the pathways of circulation and re-absorption of CSF. SEARCH STRATEGY: Pediatric, Neurosurgical and General Medical Journals were handsearched from 1976, as well as the Medline database. Personal contacts were used. SELECTION CRITERIA: One randomised trial evaluated intraventricular streptokinase in infants developing post-hemorrhagic ventricular dilatation. DATA COLLECTION AND ANALYSIS: Details of patient selection, patient allocation and the interventions were extracted. The end-points examined were: ventriculoperitoneal shunt, death, meningitis, and secondary hemorrhage. MAIN RESULTS: When intraventricular streptokinse was compared with conservative management of post hemorrhagic ventricular dilatation, the numbers of deaths and babies with shunt dependence were identical in both groups. No information on the effect of intraventricular streptokinase on disability is available. There is cause for concern about meningitis and secondary intraventricular hemorrhage but numbers are insufficient to quantify the risks. REVIEWER'S CONCLUSIONS: Intraventricular fibrinolytic therapy with streptokinase, given when post-hemorrhagic ventricular dilatation is established, cannot be recommended for neonates following IVH. A conservative approach with CSF drainage applied only to symptomatic raised intracranial pressure seems appropriate. PMID- 10796212 TI - Kinesthetic stimulation for treating apnea in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: Main question: in preterm infants with apnea, does the use of kinesthetic stimulation lead to clinically important reductions in clinical apnea and bradycardia (>50% reduction in number of episodes), use of mechanical ventilation (IPPV) or continuous positive airways pressure (CPAP), and neurodevelopmental disability, without clinically important side effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences, symposia proceedings, expert informants, and journal handsearching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random patient allocation in which kinesthetic stimulation in preterm infants was compared to placebo or no treatment for apnea of prematurity were included. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used with separate evaluation of trial quality, data extraction by both authors and synthesis of data using relative risk and weighted mean difference. As all three trials were crossover trials, the data were extracted from all exposure periods and combined where appropriate. Measures of severity of apnea as well as the response to treatment were consistent with an evaluation of 'clinical apnea', as defined by the American Academy of Pediatrics (Nelson 1978). MAIN RESULTS: Three crossover studies (Korner 1978, Tuck 1982 and Jirapaet 1993) were identified that compared a form of kinesthetic stimulation to control for the treatment of apnea of prematurity. Clinically significant apnea: None of the three studies showed an important reduction (>50%) in clinical apnea. Using a lower threshold (>25%), the study by Korner 1978 found less apnea and bradycardia in infants receiving kinesthetic stimulation. Tuck 1982 demonstrated reductions in the frequencies of apneas (> 12 seconds) associated with bradycardia (< 100 bpm), apneas associated with hypoxia (TcP02 < 50 mmHg), and apneas requiring stimulation in infants on the rocking bed. Individual patient data were not available from the author to determine if there was an important reduction in clinical apnea. No outcome could be extracted from the study by Jirapaet 1993 that was consistent with the definition of clinically important apnea. Other events: No significant differences were found in the incidence of infants requiring resuscitation, IPPV, CPAP or respiratory stimulants whilst receiving treatment. Adverse events such as death, intraventricular hemorrhage and neurodevelopmental follow up were not reported. REVIEWER'S CONCLUSIONS: There is insufficient evidence to recommend kinesthetic stimulation as treatment for clinically significant apnea of prematurity. Previous reviews have suggested that kinesthetic stimulation is not effective at preventing apnea of prematurity (Henderson-Smart and Osborn 1998) and is not as effective as theophylline at treating clinically significant apnea of prematurity (Osborn and Henderson-Smart 1998). PMID- 10796213 TI - Endotracheal intubation at birth for preventing morbidity and mortality in vigorous, meconium-stained infants born at term. AB - BACKGROUND: On the basis of evidence from non-randomised studies, it has been recommended that all babies born through thick meconium should have their tracheas intubated so that suctioning of their airways can be performed. The aim is to reduce the incidence and severity of meconium aspiration syndrome. However, for term babies who are vigorous at birth endotracheal intubation may be both difficult and unnecessary. OBJECTIVES: To determine if endotracheal intubation and suction of the airways at birth in vigorous term meconium-stained babies is more beneficial than routine resuscitation including aspiration of the oro pharynx. SEARCH STRATEGY: The search was made from Oxford Database of Perinatal Trials, the Neonatal Trials Registry of the Cochrane Neonatal Collaborative Review Group and information obtained from knowledgeable practising neonatologists. SELECTION CRITERIA: Randomised trials which compared a policy of routine vs no (or selective) use of endotracheal intubation and aspiration in the immediate management of vigorous term meconium-stained babies at birth. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, meconium aspiration syndrome, other respiratory conditions, pneumothorax, need for oxygen supplementation, stridor, convulsions and hypoxic-ischaemic encephalopathy were abstracted and analysed using Revman 3.1.1. MAIN RESULTS: Four randomised controlled trials of endotracheal intubation at birth in vigorous term meconium-stained babies were identified. Meta-analysis of these trials does not support routine use of endotracheal intubation at birth in vigorous meconium stained babies to reduce mortality, meconium aspiration syndrome, other respiratory symptoms or disorders, pneumothorax, oxygen need, stridor, HIE and convulsions. REVIEWER'S CONCLUSIONS: Routine endotracheal intubation at birth in vigorous term meconium-stained babies has not been shown to be superior to routine resuscitation including oro-pharyngeal suction. This procedure cannot be recommended for vigorous infants until more research is available. PMID- 10796214 TI - Kinesthetic stimulation versus theophylline for apnea in preterm infants. AB - BACKGROUND: Apnea of prematurity may lead to hypoxemia and bradycardia requiring resuscitative measures being instituted. Many treatments have been used in infants with apnea of prematurity, such as theophylline. Kinesthetic stimulation, which uses various forms of oscillating mattress, might also prevent apnea without using a standard drug such as theophylline. OBJECTIVES: Main question: in preterm infants, how does kinesthetic stimulation compare with methylxanthine therapy in the treatment of apnea of prematurity. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants and journal handsearching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random patient allocation, in which kinesthetic stimulation was compared to methylxanthine therapy for apnea of prematurity, were eligible. No trials were excluded from the review that met these criteria. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used with separate evaluation of trial quality, data extraction by both authors and synthesis of data using relative risk and weighted mean difference. MAIN RESULTS: A single small study of 20 infants (Saigal 1986) demonstrated a significant benefit to the infants receiving theophylline compared to those on an oscillating water bed in terms of mean rates of clinically important apnea (apnea > 14 seconds and bradycardia < 100, and cyanosis or receiving stimulation). There were no significant differences in adverse effects (death, sleep states, the Albert Einstein Neurobehavioural Index, adverse neurological outcomes, and the Bayley Mental Development Index at six and 12 months), although the infants on the OWB had a higher psychomotor index at six but not 12 months. There were some differences between the groups in incidence and severity of respiratory distress syndrome, and baseline apnea rates. REVIEWER'S CONCLUSIONS: The results of this review should be treated with caution. Theophylline has been shown in one small study to be superior to kinesthetic stimulation at treating clinically important apnea of prematurity. There are currently no clear research questions regarding the comparison of methylxanthines and kinesthetic stimulation to treat apnea of prematurity. PMID- 10796215 TI - Inhaled nitric oxide for respiratory failure in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine whether, in preterm newborn infants who have hypoxic respiratory failure, treatment with inhaled nitric oxide improves oxygenation and reduces the rates of death, or adverse neurodevelopmental outcome. SEARCH STRATEGY: Electronic and hand searching of pediatric/neonatal literature and personal data files. SELECTION CRITERIA: Randomized and quasi randomized studies in preterm infants with hypoxic respiratory failure. Administration of inhaled nitric oxide. Clinically relevant outcomes, including death, oxygenation, intraventricular haemorrhage. DATA COLLECTION AND ANALYSIS: One randomized controlled trial of nitric oxide therapy was found in preterm infants with a high risk of developing bronchopulmonary dysplasia (Subhedar 1997). MAIN RESULTS: No significant effect of inhaled nitric oxide on any outcome variable was found. In particular there was no effect on the primary outcome variable of death or bronchopulmonary dysplasia. REVIEWER'S CONCLUSIONS: There is currently no published information to support the use of inhaled nitric oxide in preterm infants. Preterm infants should not be treated with inhaled nitric oxide except in the context of prospective, randomized controlled trials. PMID- 10796216 TI - Antibiotic prophylaxis for cesarean section. AB - BACKGROUND: The single most important risk factor for postpartum maternal infection is Cesarean delivery. OBJECTIVES: The objective of this review was to assess the effects of prophylactic antibiotic treatment on infectious complications in women undergoing Cesarean delivery. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials comparing antibiotic prophylaxis or no treatment for both elective and non-elective Cesarean section. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Sixty-six trials were included. Use of prophylactic antibiotics in women undergoing Cesarean section substantially reduced the incidence of episodes of fever, endometritis, wound infection, urinary tract infection and serious infection after Cesarean section. The reduction in the risk of endometritis with antibiotics was similar across different patient groups. The relative risk for elective Cesarean section was 0.24, 95% confidence interval 0.11 to 0.48. The relative risk for non-elective Cesarean section was 0.30, 95% confidence interval 0.25 to 0.35. The relative risk for undefined or all patients together was 0.29, 95% confidence interval 0.26 to 0.33. Despite the large number of trials, different populations and different antibiotic regimens, there was no statistically significant heterogeneity. REVIEWER'S CONCLUSIONS: The reduction of endometritis by two thirds to three quarters justifies a policy of administering prophylactic antibiotics to women undergoing elective or non-elective Cesarean section. PMID- 10796217 TI - Patterns of routine antenatal care for low-risk pregnancy. AB - BACKGROUND: It has been suggested that reduced antenatal care packages or prenatal care managed by providers other than obstetricians for low risk women can be as effective as standard models of antenatal care. OBJECTIVES: The objective of this review was to assess the effects of antenatal care programmes for low-risk women. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, reference lists of articles and we also contacted researchers in the field. Date of last search: April 1999. SELECTION CRITERIA: Randomised trials comparing programmes of antenatal care with varied frequency and timing of the visits and different types of care providers. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers independently. Study authors were contacted for additional information and they were provided with the final version of the review. MAIN RESULTS: Nine trials involving over 25000 women were included. Six trials evaluated the number of visits and three trials evaluated the type of care provider. Most trials were of acceptable quality. Moderate reduction in the number of visits was not associated with an increase in any of the negative perinatal outcomes reviewed. However, trials from developed countries suggest that women can be less satisfied with the reduced number of visits and feel that their expectations with care are not fulfilled. Antenatal care provided by a midwife/general practitioner was associated with improved perception of care by women. Clinical effectiveness of midwife/general practitioner managed care was similar to that of obstetrician/gynaecologist led shared care. REVIEWER'S CONCLUSIONS: It appears that a moderate reduction in the number of antenatal care visits with an increased emphasis on the content of the visits could be implemented without any increase in adverse biological perinatal outcomes. Women can be less satisfied with reduced visits. While clinical effectiveness seemed similar, women appeared to be slightly more satisfied with midwife/general practitioner managed care compared to obstetrician/gynaecologist led shared care. PMID- 10796218 TI - Antenatal education for self-diagnosis of the onset of active labour at term. AB - BACKGROUND: A specific program designed to teach women to recognise active labour may be beneficial through potentially decreasing the incidence of early admission to hospital, increasing women's confidence and decreasing their anxiety. OBJECTIVES: The objective of this review was to assess the effects of teaching pregnant women specific criteria for self-diagnosis of active labour onset in term pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials comparing a structured antenatal education intervention for the identification of symptoms for self-diagnosis of active labour with usual care. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: One study involving 245 women was included. Method of randomisation was unclear and 15% of the sample was lost to follow-up in this trial. A specific antenatal education program was associated with a reduction in the mean number of visits to the labour suite before the onset of labour (weighted mean difference 0. 29, 95% confidence interval -0.47 to -0.11). It is unclear whether this resulted in fewer women being sent home because they were not in labour. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of a specific set of criteria for self-diagnosis of active labour. PMID- 10796219 TI - Caregivers' use of strict criteria for diagnosing active labour in term pregnancy. AB - BACKGROUND: Application of specific criteria for diagnosis of active labour as part of a labour assessment program aims to differentiate more accurately between latent and active phases of labour. OBJECTIVES: The objective of this review was to assess the effects of the use of specific criteria by caregivers in diagnosing active labour in term pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: January 1998. SELECTION CRITERIA: Randomised trials comparing caregivers' application of strict diagnostic criteria for active labour with routine care. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: One study of 209 women was included. The trial was of excellent quality. Women who experienced early labour assessment were less likely to receive intrapartum oxytocics than women who received standard care (odds ratio 0.45, 95% confidence interval 0.25 to 0.80) and analgesia (odds ratio 0.36, 95% confidence interval 0.16 to 0.78). They reported higher levels of control during labour and birth (weighted mean difference 16.00, 95% confidence interval 7.52 to 24.48). There were no differences detected for rate of caesarean section and other important measures of maternal and neonatal outcome. REVIEWER'S CONCLUSIONS: Early labour assessment (which includes use of specific criteria for diagnosis of active labour) may have some positive outcomes for women at term pregnancy. PMID- 10796220 TI - Magnesium supplementation in pregnancy. AB - BACKGROUND: Many women, especially those from disadvantaged backgrounds, have intakes of magnesium below recommended levels. Magnesium supplementation during pregnancy may be able to reduce fetal growth retardation and pre-eclampsia, and increase birthweight. OBJECTIVES: The objective of this review was to assess the effects of magnesium supplementation during pregnancy on maternal, neonatal and paediatric outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi randomised trials of dietary magnesium supplementation during pregnancy. DATA COLLECTION AND ANALYSIS: Suitability for inclusion and methodological quality were separately assessed by each reviewer. Data were independently extracted by two reviewers. MAIN RESULTS: Six trials involving 2637 women were included. Only one of these trials was judged to be of high quality. Compared with placebo, oral magnesium treatment from before the 25th week of gestation was associated with a lower incidence of preterm birth (odds ratio 0.71, 95% confidence interval 0.52 to 0.95). There was also less maternal hospitalisation during pregnancy, fewer cases of antepartum haemorrhage, a lower incidence of low birthweight and small for gestational age infants. Poor quality trials are likely to have resulted in a bias favouring magnesium supplementation. REVIEWER'S CONCLUSIONS: There is not enough high quality evidence to show that dietary magnesium supplementation during pregnancy is beneficial. PMID- 10796221 TI - Induction of labour for suspected fetal macrosomia. AB - BACKGROUND: Suspected macrosomic fetuses are usually induced in order to reduce the risk of difficult operative delivery. OBJECTIVES: The objective of this review was to assess the effects of a policy of labour induction for suspected fetal macrosomia on method of delivery and maternal or perinatal morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials of induction of labour for suspected fetal macrosomia in non diabetic women. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done independently by two reviewers. Study authors were contacted for additional information. MAIN RESULTS: Two trials involving 313 women were included. Compared to expectant management, induction of labour for suspected macrosomia did not reduce the risk of caesarean section (odds ratio 0.85, 95% confidence interval 0.50 to 1.46) or instrumental delivery (odds ratio 0.98, 95% confidence interval 0.48 to 1.98). Perinatal morbidity was similar between groups. REVIEWER'S CONCLUSIONS: Induction of labour for suspected fetal macrosomia in non-diabetic women did not appear to alter the risk of maternal or neonatal morbidity. PMID- 10796222 TI - Magnesium for preventing preterm birth after threatened preterm labour. AB - BACKGROUND: Magnesium therapy is sometimes continued after an episode of threatened preterm labour in an attempt to prevent the onset of further preterm contractions. OBJECTIVES: The objective of this review was to assess the effects of magnesium maintenance therapy on preventing preterm birth after threatened preterm labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised controlled trials of magnesium therapy given to women after threatened preterm labour. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done independently by two reviewers. MAIN RESULTS: Two trials were included. Both were of poor quality and neither study included any long-term follow up of infants. The incidence of preterm birth or perinatal mortality was not reduced by magnesium maintenance therapy compared to alternative therapy or no treatment. REVIEWER'S CONCLUSIONS: There is not enough evidence to show that magnesium maintenance therapy is effective in preventing preterm birth after an episode of threatened preterm labour. PMID- 10796223 TI - Vaginal misoprostol for cervical ripening and labour induction in late pregnancy. AB - BACKGROUND: Although not yet registered for such use, misoprostol has been widely used for obstetric and gynaecological indications, such as induction of abortion and of labour. OBJECTIVES: The objective of this review was to assess the effects of vaginal misoprostol for third trimester cervical ripening or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and reference lists of articles were searched. SELECTION CRITERIA: Randomised trials comparing vaginal misoprostol with other methods of cervical ripening or labour induction, placebo or no treatment in women due for third trimester induction of labour. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done by both reviewers. MAIN RESULTS: Twenty-six studies were included. Compared to placebo, misoprostol was associated with increased cervical ripening (relative risk of unfavourable or unchanged cervix after 12 to 24 hours with misoprostol 0.20, 95% confidence interval 0.07 to 0.61). It was also associated with a reduced need for oxytocin (relative risk 0.47, 95% confidence interval 0.23 to 0.96). Misoprostol was more effective than prostaglandin E2 for labour induction (relative risk of failure to achieve vaginal delivery in 24 hours 0.70, 95% confidence interval 0.62 to 0.79). Oxytocin augmentation was used less often with misoprostol than with prostaglandin E2 (relative risk 0.64, 95% confidence interval 0.58 to 0.71). Uterine hyperstimulation and meconium stained liquor were more common with misoprostol than with prostaglandin E2. Lower doses of misoprostol compared to higher doses did not show significant differences except for more need for oxytocin augmentation and less uterine hyperstimulation without fetal heart rate changes. REVIEWER'S CONCLUSIONS: Vaginal misoprostol appears to be more effective in inducing labour than conventional methods of cervical ripening and labour induction. The apparent increase in uterine hyperstimulation is of concern. The studies were not large enough to exclude the possibility of rare but serious adverse effects. PMID- 10796224 TI - Amnioinfusion for preterm rupture of membranes. AB - BACKGROUND: Preterm rupture of membranes places a fetus at risk of cord compression and amnionitis. Amnioinfusion aims to prevent or relieve umbilical cord compression by infusing a solution into the uterine cavity. OBJECTIVES: The objective of this review was to assess the effects of amnioinfusion for preterm rupture of membranes on maternal and perinatal outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials of amnioinfusion compared to no amnioinfusion in women with preterm rupture of membranes. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: One trial of 66 women was included. It had some methodological flaws. No significant differences between amnioinfusion and no amnioinfusion were detected for caesarean section (relative risk 0.32, 95% confidence interval 0.07 to 1.40); low Apgar scores (relative risk 0.28, 95% confidence interval 0.03 to 2.33) or neonatal death (relative risk 0.55, 95% confidence interval 0.05 to 5.77). In the amnioinfusion group, the number of severe fetal heart rate decelerations per hour during the first stage of labour were reduced (weighted mean difference -1.20, 95% confidence interval -1.83 to 0.57). These outcomes are consistent with those found in the Cochrane review on amnioinfusion for cord compression. REVIEWER'S CONCLUSIONS: There is not enough evidence concerning the use of amnioinfusion for preterm rupture of membranes. PMID- 10796225 TI - Symphysis-fundal height measurement in pregnancy. AB - BACKGROUND: In many settings, symphysis-fundal height measurement has replaced clinical assessment of fetal size by abdominal palpation because the latter has been reported to perform poorly. OBJECTIVES: The objective of this review was to assess the effects of routine use of symphysis-fundal height measurements (tape measurement of the distance from the pubic symphysis to the uterine fundus) during antenatal care on pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. SELECTION CRITERIA: Acceptably controlled trials comparing symphysis-fundal height measurement with assessment by abdominal palpation alone. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: One trial involving 1639 women was included. No differences were detected in any of the outcomes measured. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of symphysis-fundal height measurements during antenatal care. PMID- 10796226 TI - Caregiver support for postpartum depression. AB - BACKGROUND: Supportive relationships during the perinatal period may enhance a mother's feeling of wellbeing and control. Support to women during labour and after birth has shown benefits and this may also be the case for mothers with postpartum depression. OBJECTIVES: The objective of this review was to assess the effect of professional and/or social support interventions for the treatment of postpartum depression. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi randomised trials comparing additional support from caregivers with usual forms of care in the postpartum period, in women who were clinically depressed in the six months after giving birth. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by both reviewers. Study authors were contacted for additional information. MAIN RESULTS: Two studies involving 137 women were included. There is potential for bias in at least one study, due to large numbers of women refusing to take part in the trial as well as significant losses to follow-up during the trial. Treatment of postpartum depression with support was associated with a reduction in depression at 25 weeks after giving birth (odds ratio 0.34, 95% confidence intervals 0.17 to 0.69). REVIEWER'S CONCLUSIONS: There is some indication that professional and/or social support may help in the treatment of postpartum depression. The types of support should be investigated to assess which models are most effective. PMID- 10796227 TI - Continuous versus interrupted sutures for perineal repair. AB - BACKGROUND: Millions of women worldwide undergo perineal repair after childbirth and the type of repair may have an impact on pain and healing. OBJECTIVES: The objective of this review was to assess the effects of continuous subcuticular versus interrupted transcutaneous sutures on women following perineal repair. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. Date of last search: June 1999. SELECTION CRITERIA: Randomised trials comparing continuous subcuticular versus interrupted transcutaneous sutures for the closure of perineal skin after vaginal delivery. DATA COLLECTION AND ANALYSIS: Trial quality was assessed independently by two reviewers. Data were extracted by one reviewer and checked by the second reviewer. Study authors were contacted for additional information. MAIN RESULTS: Four studies involving 1864 women were included. The trials were heterogeneous with respect to operator skill and training. Compared with interrupted sutures, a continuous subcuticular suture technique of perineal repair was associated with less pain for up to 10 days post partum (odds ratio 0.68, 95% confidence interval 0.53 to 0.86). No differences were seen in the need for analgesia, need for resuturing of the wound or in dyspareunia. Based on one trial only, there was no difference in long-term pain and failure to resume pain-free intercourse within three months of the birth. The continuous technique was associated with less need for the removal of sutures. REVIEWER'S CONCLUSIONS: The continuous subcuticular technique of perineal repair may be associated with less pain in the immediate postpartum period than the interrupted suture technique. The long-term effects are less clear. PMID- 10796228 TI - Interventions for promoting smoking cessation during pregnancy. AB - BACKGROUND: Smoking remains one of the few potentially preventable factors associated with low birthweight, very preterm birth and perinatal death. OBJECTIVES: The objective of this review was to assess the effects of smoking cessation programs implemented during pregnancy on the health of the fetus and infant, on the mother and on the family. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: Randomised and quasi randomised trials of smoking cessation programs implemented during pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Forty-four trials were identified: 37 trials including 16,916 women provided data on smoking cessation and/or perinatal outcomes, as did one cluster-randomised trial including 3000 women. Over 800 women were included in trials of smoking relapse prevention. There was substantial variation in the intensity of the intervention and the extent of reminders and reinforcement through pregnancy. Based on 34 trials there was a significant reduction in smoking in the intervention groups (odds ratio 0.53, 95% confidence interval 0. 47 to 0.60), an absolute difference of 6.4% women continuing to smoke. The eight trials with validated smoking cessation, a high intensity intervention and a high quality score had an odds ratio of 0.53, 95% confidence interval 0.44 to 0.63 and an absolute difference in continued smoking of 8.1%. The subset of trials with information on fetal outcome revealed a reduction in low birthweight (odds ratio 0.80, 95% confidence interval 0.67 to 0.95), a reduction in preterm birth (odds ratio 0.83, 95% confidence interval 0.69 to 0. 99) and an increase in mean birthweight of 28g (95% confidence interval 9 to 49). There were no differences in very low birthweight or perinatal mortality. Five trials of smoking relapse prevention showed no significant difference. The single large cluster-randomised trial showed no evidence of a decrease in continued smoking or adjusted mean birthweight. REVIEWER'S CONCLUSIONS: Smoking cessation programs in pregnancy appear to reduce smoking, low birthweight and preterm birth, but no effect was detected for very low birthweight or perinatal mortality. PMID- 10796229 TI - Periconceptional supplementation with folate and/or multivitamins for preventing neural tube defects. AB - BACKGROUND: Neural tube defects arise during the development of the brain and spinal cord. OBJECTIVES: The objective of this review was to assess the effects of increased consumption of multivitamins or folate on the prevalence of neural tube defects before pregnancy and in the first two months of pregnancy (periconceptionally). SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised and quasi randomised trials comparing periconceptional supplementation by multivitamins with placebo, folate with placebo, or multivitamins with folate; different dosages of multivitamins or folate; prepregnancy dietary advice and counselling in primary care settings to increase the consumption of folate-rich foods, or folate-fortified foods, with standard care; increased intensity of information provision with standard public health dissemination. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Four trials involving 6425 women were included. The trials all addressed the question of supplementation and they were of variable quality. No dissemination trials were identified. Periconceptional folate supplementation reduced the incidence of neural tube defects (odds ratio 0.28, 95% confidence interval 0.15 to 0.53). Folate supplementation did not significantly increase spontaneous abortion, ectopic pregnancy or stillbirth, although there was a possible increase in multiple gestation. Multivitamins alone were not associated with prevention of neural tube defects and did not produce preventive effects when given with folate. REVIEWER'S CONCLUSIONS: Periconceptional folate supplementation has a strong protective effect against neural tube defects. Information about folate should be made more widely available throughout the health and education systems. Women whose fetuses or babies have neural tube defects should be offered continuing folate supplementation. The benefits and risks of fortifying basic food stuffs, such as flour, with added folate remain unresolved. PMID- 10796230 TI - Topical umbilical cord care at birth. AB - BACKGROUND: Umbilical cord infection caused many neonatal deaths before aseptic techniques were used. OBJECTIVES: The objective of this review was to assess the effects of topical cord care in preventing cord infection, illness and death. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register (Cochrane Library issue 4, 1997) and Medline. We also contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of topical cord care compared with no routine care, and comparisons between different forms of care. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Ten studies were included, all from developed countries. No systemic infections or deaths were observed in any of the studies reviewed. Cord and other skin infections within six weeks of observation were not affected by use of antiseptics. There was a trend to reduced colonization with antibiotics compared to antiseptics and no treatment. Antiseptics prolonged the time to cord separation. Use of antiseptics was associated with a reduction in maternal concern about the cord. REVIEWER'S CONCLUSIONS: Simply keeping the cord clean appears to be as effective and safe as using antibiotics or antiseptics. PMID- 10796231 TI - Antibiotics for preterm premature rupture of membranes. AB - OBJECTIVES: The aim of the review was to evaluate the effectiveness and the immediate and long-term safety of the effects of administering antibiotics to women with preterm prelabour rupture of membranes on maternal infectious morbidity, fetal and neonatal morbidity and mortality, and longer term childhood development. SEARCH STRATEGY: All randomized trials identified using the search strategy described by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All trials which reported clinically relevant outcomes (as opposed to laboratory data) were included. DATA COLLECTION AND ANALYSIS: Data were extracted from each report without any blinding of either the results or the treatments which women received. Unpublished data were sought from a number of authors. MAIN RESULTS: Antibiotic treatment following pPROM is effective at prolonging pregnancy and reducing maternal and neonatal infectious morbidity. There is no statisically proven benefit that their use improves neonatal mortality and morbidity in the short or long term. REVIEWER'S CONCLUSIONS: There are insufficient data to recommend routine prescription of antibiotics in this clinical situation. Research into this area should continue. PMID- 10796232 TI - Calcium supplementation during pregnancy for preventing hypertensive disorders and related problems. AB - BACKGROUND: Calcium supplementation may prevent high blood pressure through a number of mechanisms and may help to prevent preterm labour. OBJECTIVES: The objective of this review was to assess the effects of calcium supplementation during pregnancy on hypertensive disorders of pregnancy and related maternal and child adverse outcomes. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register and we contacted study authors. SELECTION CRITERIA: Randomised trials comparing at least one gram daily of calcium during pregnancy compared to placebo. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. Data extraction was carried out independently by two reviewers. MAIN RESULTS: Nine studies were included, all of good quality. There was a modest reduction in high blood pressure with calcium supplementation (relative risk 0.80, 95% confidence interval 0.73 to 0.88). The effect was greatest for women at high risk of hypertension (relative risk 0.35, 95% confidence interval 0.21 to 0.57) and those with low baseline dietary calcium (relative risk 0.49, 95% confidence interval 0.38 to 0.62). There was also a modest reduction in the risk of pre-eclampsia with calcium supplementation (relative risk 0. 72, 95% confidence interval 0.60 to 0.86). The effect was greatest for women at high risk of hypertension (relative risk 0.22, 95% confidence interval 0.11 to 0.43) and those with low baseline calcium intake (relative risk 0.32, 95% confidence interval 0.21 to 0.49). There was no overall effect on the risk of preterm delivery, although there was a reduction in risk amongst women at high risk of hypertension (relative risk 0.42, 95% confidence interval 0.23 to 0. 78). There was no evidence of any effect of calcium supplementation on stillbirth or death before discharge from hospital. REVIEWER'S CONCLUSIONS: Calcium supplementation appears to be beneficial for women at high risk of gestational hypertension and in communities with low dietary calcium intake. Optimum dosage requires further investigation. PMID- 10796233 TI - Abdominal decompression in normal pregnancy. AB - BACKGROUND: Abdominal decompression was developed as a means of pain relief during labour. It has also been used for complications of pregnancy, and in healthy pregnant women in an attempt to improve fetal wellbeing and intellectual development. OBJECTIVES: The objective of this review was to assess the effects of prophylactic abdominal decompression on admission for pre-eclampsia, fetal growth, perinatal morbidity and mortality and childhood development. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials comparing abdominal decompression with dummy decompression or no treatment in healthy pregnant women. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Three studies were included. There was no difference between the abdominal decompression groups and the control groups for low birth weight (relative risk 0.69, 95% confidence interval 0.27 to 1.77) and perinatal mortality (relative risk 2.47, 95% confidence interval 0.77 to 7.92). There were no differences in admission for pre-eclampsia, Apgar score and childhood development. REVIEWER'S CONCLUSIONS: There is no evidence to support the use of abdominal decompression in normal pregnancies. Future research should be directed towards the use of abdominal decompression during labour, and during complicated pregnancies. PMID- 10796234 TI - Hands/knees posture in late pregnancy or labour for fetal malposition (lateral or posterior). AB - BACKGROUND: Lateral and posterior position of the fetal presenting parts may be associated with more painful, prolonged or obstructed labour and difficult delivery. It is possible that maternal posture may influence fetal position. OBJECTIVES: The objective of this review is to assess the effects of adopting a hands and knees maternal posture in late pregnancy when the presenting part of the fetus is in a lateral or posterior position. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials of hands and knees maternal posture compared to other postures. DATA COLLECTION AND ANALYSIS: Both reviewers assessed trial eligibility and quality. MAIN RESULTS: One trial involving 100 women was included. Four different postures (four groups of 20 women) were combined for the comparison with the control group of 20 women. Lateral or posterior position of the presenting part of the fetus was less likely to persist following 10 minutes in the hands and knees position compared to a sitting position (relative risk 0.25, 95% confidence interval 0.17 to 0.37). REVIEWER'S CONCLUSIONS: Hands and knees maternal posture for lateral or posterior fetal presentation appears to result in short term effects ion fetal position. No other perinatal or maternal outcomes were reported. There is not enough evidence to evaluate the effectiveness of a hands and knees maternal posture when the fetal presenting part is lateral or posterior, on clinically important outcomes. PMID- 10796235 TI - Piracetam for fetal distress in labour. AB - BACKGROUND: Piracetam is thought to promote the metabolism of brain cells when they are hypoxic. It has been used to prevent adverse effects of fetal distress. OBJECTIVES: The objective of this review was to assess the effects of piracetam for suspected fetal distress in labour on method of delivery and perinatal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: February 1999. SELECTION CRITERIA: Randomised trials of piracetam compared with placebo or no treatment for suspected fetal distress in labour. DATA COLLECTION AND ANALYSIS: Both reviewers assessed eligibility and trial quality. MAIN RESULTS: One study of 96 women was included. Piracetam compared with placebo was associated with a trend to reduced need for caesarean section (relative risk 0.57, 95% confidence interval 0.32 to 1.03). There were no statistically significant differences in relative risk between the piracetam and placebo group for neonatal morbidity (measured by neonatal respiratory distress) or Apgar score. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of piracetam for fetal distress in labour. PMID- 10796236 TI - Operative versus conservative management for 'fetal distress' in labour. AB - BACKGROUND: Suspected fetal distress usually results in expedited delivery of a baby (often operatively). The potential harm to a mother and baby from operative delivery may not always be justified especially when fetal distress may be misdiagnosed. Even with a correct diagnosis it is not clear whether an operative or conservative approach is better. OBJECTIVES: The objective of this review was to assess the effects of operative management for fetal distress on maternal and perinatal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Date of last search: October 1997. SELECTION CRITERIA: Randomised trials of operative (caesarean section or expedited vaginal delivery) versus conservative management of suspected fetal distress. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done by both reviewers. MAIN RESULTS: One study of 350 women was included. This trial was carried out in 1959. There was no difference in perinatal mortality (relative risk 1.18, 95% confidence interval 0.56 to 2.48). REVIEWER'S CONCLUSIONS: There have been no contemporary trials of operative versus conservative management of suspected fetal distress. In settings without modern obstetric facilities, a policy of operative delivery in the event of meconium-stained liquor or fetal heart rate changes has not been shown to reduce perinatal mortality. PMID- 10796237 TI - Interventions for varicosities and leg oedema in pregnancy. AB - BACKGROUND: Leg oedema from venous insufficiency is not dangerous but it can cause women symptoms such as pain, feelings of heaviness, night cramps and paraesthesiae. Leg oedema can be a sign of pre-eclampsia when associated with raised blood pressure or proteinuria. OBJECTIVES: The objective of this review was to assess the effects of treatment to relieve the symptoms associated with varicosity in pregnancy and to reduce leg oedema. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. SELECTION CRITERIA: Randomised trials of any form of treatment for varicosity or leg oedema in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Three trials of three different treatments were included. A total of 115 women were involved. In one trial, two thirds of women given rutoside capsules in the last three months of pregnancy noted an improvement in symptoms compared with only one third given placebo (odds ratio 0.30, 95% confidence interval 0.12 to 0.77). They had a decrease in ankle circumference at 36 weeks' gestation after eight weeks of treatment, while women given placebo had a small increase. In one trial, women with ankle oedema had a small non-significant reduction in lower leg volume when treated with external pneumatic intermittent compression for 30 minutes. Fifty minutes immersion in water at 32 degrees Celsius resulted in greater diuresis and fall in blood pressure than 50 minutes bedrest. REVIEWER'S CONCLUSIONS: Rutosides appear to relieve symptoms of venous insufficiency in late pregnancy. However it is not known if the drug is safe in pregnancy. External pneumatic compression appears to reduce ankle swelling. Immersion in water for 50 minutes results in diuresis and fall in blood pressure. It is not known for how long these changes are sustained nor whether they are of any benefit. PMID- 10796238 TI - Antibiotic regimens for endometritis after delivery. AB - BACKGROUND: Post-partum endometritis, which is more common after cesarean section, occurs when vaginal organisms invade the endometrial cavity during labour and delivery. Antibiotic treatment is warranted. OBJECTIVES: The effect of different antibiotic regimens for the treatment of postpartum endometritis on failure of therapy and complications was systematically reviewed. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's trials register and the Cochrane Controlled Trials Register. Date of last search: August 1999. SELECTION CRITERIA: Randomised trials of different antibiotic regimens for postpartum endometritis, after cesarean section or vaginal delivery, where outcomes of treatment failure or complications were reported were selected. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by the reviewers. Comparisons were made between different types of antibiotic regimen, based on type of antibiotic and duration and route of administration. Summary relative risks were calculated. MAIN RESULTS: Forty-one trials were included. Overall the studies were methodologically poor. In the intent-to-treat analysis, fifteen studies comparing clindamycin and an aminoglycoside with another regimen showed more treatment failures with another regimen (relative risk (RR) 1.37; 95% confidence interval (CI) 1.10 - 1.70). Failures of those regimens with poor activity against penicillin resistant anaerobic bacteria were more likely (RR 1.73; 95% CI 1.14 - 2.63). In four studies that compared continued oral antibiotic therapy after intravenous therapy, no differences were found in recurrent endometritis or other outcomes. There was no evidence of difference in incidence of allergic reactions. Cephalosporins were associated with less diarrhea. REVIEWER'S CONCLUSIONS: The combination of gentamicin and clindamyin is appropriate for the treatment of endometritis. Regimens with activity against penicillin resistant anaerobic bacteria are better than those without. There is no evidence that any one regimen is associated with fewer side effects. Once uncomplicated endometritis has clinically improved with intravenous therapy, oral therapy is not needed. PMID- 10796239 TI - Cardiotocography for antepartum fetal assessment. AB - BACKGROUND: Cardiotocography is a form of fetal assessment which simultaneously records fetal heart rate, fetal movements and uterine contractions to investigate hypoxia. OBJECTIVES: The objective was to assess the effects of antenatal cardiotocography on perinatal morbidity and mortality and maternal morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (to 1 November 1998). SELECTION CRITERIA: Randomised trials comparing antenatal cardiotocography with a control group for fetal assessment. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Four studies involving 1,588 pregnancies were included. All trials were conducted on high or intermediate risk pregnancies. Antenatal cardiotocography appeared to have no significant effect on perinatal mortality or morbidity. There was a trend to an increase in perinatal deaths in the cardiotocography group (odds ratio 2.85, 95% confidence interval 0.99 to 7.12). There was no increase in the incidence of interventions such as elective caesarean section or induction of labour. The one trial which examined an effect on antenatal patient management showed a significant reduction in hospital admissions and a reduction in inpatient stay in the cardiotocography group. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of antenatal cardiotocography for fetal assessment. All of the trials included in this review date from the introduction of antenatal cardiotocography and may be difficult to relate to current practice. PMID- 10796240 TI - Continuous positive airway pressure versus theophylline for apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia which may be severe enough to require resuscitation including use of positive pressure ventilation. Theophylline stimulation of breathing and continuous positive airways pressure have been used to prevent apnea and its consequences. OBJECTIVES: The main objective was to determine in preterm infants with recurrent apnea, if treatment with CPAP compared with treatment with theophylline leads to a clinically important reduction in apnea or use of mechanical ventilation, without clinically important side effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This includes searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences, symposia proceedings, expert informants and journal hand searching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random allocation to CPAP or theophylline in preterm infants with clinical recurrent apnea/bradycardia were eligible. DATA COLLECTION AND ANALYSIS: Data were extracted using standard methods of the Cochrane Collaboration and its Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author and synthesis of data using relative risk. MAIN RESULTS: The use of mask CPAP is associated with a higher treatment failure rate as measured by less than a 50% reduction in apnea or use of the alternative treatment [RR 2.89 (95% CI 1.12,7.47); RD 0.42 (95% CI 0. 11, 0.74)]. For every 2.4 infants (95% CI 1.4, 9.5) treated with mask CPAP rather than theophylline, there results one treatment failure. In the mask CPAP group there is more use of IPPV [RR 3.09 (1.42,6.70; RD 0.58 (95% CI 0.30, 0.86). For every 1.7 infants (95% CI 1.2, 3.3) treated with mask CPAP rather than theophylline, one infant is intubated for IPPV. In the mask CPAP group, there are trends towards more deaths in the first year, and in death or major disability in survivors at follow up, which do not reach significance. There are no differences in rates of necrotizing enterocolitis or major disability in survivors at follow up. REVIEWER'S CONCLUSIONS: Theophylline is more effective than mask CPAP for preterm infants with apnea. Since CPAP is no longer administered by mask, the results of this review have limited importance for current clinical practice. Further evaluation of the benefits and harms of CPAP vs theophylline for preterm infants with apnea requires further trials in which CPAP is administered by current methods. PMID- 10796241 TI - D-Penicillamine for preventing retinopathy of prematurity in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To answer the question: Among very low birth weight infants, what is the effect of prophylactic administration of d-penicillamine on the incidence of acute ROP or severe ROP, and side effects including death? SEARCH STRATEGY: Searches were made of multiple electronic databases, previous reviews including cross references, abstracts, conference/symposia proceedings, and expert informants. SELECTION CRITERIA: Randomized or quasi-randomized controlled trials that administered d-penicillamine to infants less than 2000g birth weight within the day following birth were considered relevant to this review. Additional case series were examined for potential side effects. DATA COLLECTION AND ANALYSIS: Data on clinical outcomes were excerpted by 3 reviewers independently, and consensus reached. Data analysis was conducted according to the standards of the Neonatal Cochrane Review Group. MAIN RESULTS: Two randomized trials on the effects on ROP were identified. When combined, they showed a significantly lower incidence of acute ROP in the treated infants, relative risk of 0.09, 95% CI [0.01,0.71]. Severe stages of ROP could not be analyzed. There was no effect on death rates, relative risk 0.99 95% CI [0.70,1.39]. No side effects were reported, and follow up at one year revealed no significant differences in spasticity or developmental delay, although there were more rehospitalizations among the controls. In other reports of using d-penicillamine in over 140 infants for hyperbilirubinemia, skin rashes were reported in 2 infants and one had vomiting that may have been related. REVIEWER'S CONCLUSIONS: D-penicillamine is unlikely to affect survival, and may reduce the incidence of acute ROP among survivors. Studies to date justify further investigation of this drug in a broader population; careful attention to possible side effects is needed. PMID- 10796242 TI - Gradual versus abrupt discontinuation of oxygen in preterm or low birth weight infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: In preterm or low birth weight infants, does gradual versus abrupt discontinuation of supplemental oxygen influence mortality, retinopathy of prematurity, lung function, growth or development? SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal trials, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language. An additional literature search of the MEDLINE, EMBASE, and CINAHL databases was conducted in order to locate any trials in addition to those provided by the Cochrane Controlled Trials Register (CENTRAL/CCTR). SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which gradual weaning was compared with abrupt discontinuation of supplemental oxygen in preterm or low birth weight infants, were eligible for inclusion. DATA COLLECTION AND ANALYSIS: The methodological quality of the eligible trial was assessed independently by each author for the degree selection, performance, attrition and detection bias. Data were extracted and reviewed independently by the each author. Results were compared and differences resolved as required. Data analysis was conducted according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: The results of the one small trial of 51 infants included in this systematic review indicate a significant reduction in vascular retrolental fibroplasia (i.e. severe ROP) for infants weaned gradually from high oxygen concentrations compared with abrupt discontinuation (RR 0.22, 95% CI 0.07-0.68). This finding was independent of the duration of oxygen therapy. REVIEWER'S CONCLUSIONS: The results of this systematic review provide additional evidence linking routine exposure to high ambient oxygen in the early neonatal period to the development of ROP in preterm/LBW infants. However, due to small numbers and historical oxygen monitoring techniques, they provide little assistance to clinicians with regard to the most appropriate method of oxygen weaning, gradual or abrupt, in modern neonatal care settings. PMID- 10796243 TI - Early versus late discontinuation of oxygen in preterm or low birth weight infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: In preterm or low birth weight infants, does early versus late weaning from supplementary oxygen influence mortality, retinopathy of prematurity, lung function, growth or development? SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, MEDLINE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal handsearching mainly in the English language. An additional literature search of the MEDLINE, EMBASE, and CINAHL databases was conducted in order to locate any trials in addition to those provided by the Cochrane Controlled Trials Register (CENTRAL/CCTR). SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which early weaning was compared with late discontinuation of supplemental oxygen in preterm or low birth weight infants, were eligible for inclusion. DATA COLLECTION AND ANALYSIS: The methodological quality of the one eligible trial was assessed independently by each author for the degree of selection, performance, attrition and detection bias. Data regarding clinical outcomes including mortality, retinopathy of prematurity, and long term growth and development were extracted and reviewed independently by each author. Results were compared and differences resolved as required. Data analysis was conducted according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: In the single eligible trial of 99 infants with birthweights less than 1650g, there were no significant differences in neonatal death rates or retrolental fibroplasia (any grade or severe) for all infants, or among infants with birth weights of less than 1000g. No other outcome measures specified a priori as clinically meaningful were reported in enough detail or with satisfactory follow-up rates to include in the analysis (early death; chronic lung disease; and long term growth, development, lung or visual function). REVIEWER'S CONCLUSIONS: The results of this systematic review do not provide strong evidence for either the benefits or harms of early oxygen weaning in preterm/LBW infants. Future research should be directed toward addressing the question of what are the most appropriate target levels of oxygenation, in both the early and late neonatal periods, rather than whether oxygen should be weaned early or late. PMID- 10796244 TI - Extubation from low-rate intermittent positive airways pressure versus extubation after a trial of endotracheal continuous positive airways pressure in intubated preterm infants. AB - BACKGROUND: Failure of extubation and subsequent reintubation may result in additional stress and trauma to the premature infant. Testing infants about to be extubated with a period of endotracheal CPAP has been suggested as a method of demonstrating readiness for extubation. However, this process has been criticized as increasing the neonate's work of breathing and perhaps increasing the likelihood of extubation failure. OBJECTIVES: In premature infants having their endotracheal tube removed, is direct extubation from low rate intermittent positive pressure ventilation (IPPV) more successful than that following a period of endotracheal continuous positive airway pressure (CPAP)? SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group as outlined in the Cochrane Library was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences, symposia proceedings, expert informants and journal hand searching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random allocation of premature infants to endotracheal CPAP or direct extubation following a period of IPPV were included. DATA COLLECTION AND ANALYSIS: Data were extracted using standard methods of the Cochrane Collaboration and its Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author and synthesis of data using relative risk. MAIN RESULTS: Direct extubation from low rate ventilation is associated with a trend to increased chance of successful extubation when compared to extubation after a period of endotracheal CPAP, RR 0.45 (0.19,1.07), RD -0.103 (-0.200,-0.006), NNT 10 (5,167). When only truly randomized trials are considered, this result becomes both statistically significant and clinically important, RR 0.10 (0.01,0.78), RD -0.201 (-0.319,-0.083), NNT 5 (3, 12). Similar differences are seen for the secondary outcome, apnea. REVIEWER'S CONCLUSIONS: Preterm infants no longer requiring endotracheal intubation and IPPV should be directly extubated without a trial of ETT CPAP. PMID- 10796245 TI - Digoxin for preventing or treating neonatal respiratory distress syndrome. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effect of digoxin on clinical outcome in infants at risk of, or with, respiratory distress syndrome (RDS). SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: digoxin; limits: age groups, newborn infants; publication type, clinical trial), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, and journal handsearching in the English language. SELECTION CRITERIA: Randomized controlled trials of digoxin in either the prevention or treatment of respiratory distress syndrome are included in this overview. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes were excerpted from the trial reports by the reviewer. Data were analyzed according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Two randomized controlled trials have studied the effects of digoxin in the prevention and treatment of respiratory distress syndrome. No improvement in respiratory status or mortality was noted. Meta-analysis of the effect of digoxin given to infants at risk of or with RDS on mortality does not suggest any benefit of digoxin treatment (typical relative risk 1.27 95% CI 0.78, 2.07; typical risk difference 0.06, 95% CI -0.06, 0.17). REVIEWER'S CONCLUSIONS: Although hemodynamic disturbances play a role in the overall pathogenesis of respiratory distress syndrome, the specific contribution of early congestive heart failure (unrelated to hemodynamically significant patent ductus arteriosus) does not appear to be a significant factor in RDS. Treatment with digoxin has no proven value in infants solely affected with respiratory distress syndrome. PMID- 10796246 TI - Iron and folate supplementation in pregnancy. AB - BACKGROUND: Anaemia in pregnancy is a major health problem in many developing countries where nutritional deficiency, malaria and other parasitic infections contribute to increased maternal and perinatal mortality and morbidity. OBJECTIVES: The objective of this review was to assess the effects of routine iron and folate supplementation on haematological and biochemical parameters and on pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Study authors were also contacted. SELECTION CRITERIA: Acceptably controlled trials of routine iron and folate supplementation for pregnant women. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. Study authors were contacted for additional information. MAIN RESULTS: Eight trials involving 5449 women were included. Routine supplementation with iron or folate raised or maintained the serum iron and ferritin levels and serum and red cell folate levels. Supplementation resulted in a substantial reduction of women with a haemoglobin level below 10 or 10.5 grams in late pregnancy. Routine supplementation with iron and folate had no detectable effect on any substantive measures of either maternal or fetal outcome. REVIEWER'S CONCLUSIONS: Routine supplementation with iron and folate appears to prevent low haemoglobin at delivery. There is very little information on other outcomes for either mother or baby. There are few data derived from communities where iron and folate deficiency is common and anaemia is a serious health problem. PMID- 10796247 TI - Antibiotic prophylaxis regimens and drugs for cesarean section. AB - BACKGROUND: Prophylactic antibiotics for cesarean section have been shown to reduce the incidence of maternal postoperative infectious morbidity. Many different antibiotic regimens have been reported to be effective. OBJECTIVES: The objective of this review was to determine which antibiotic regimen is most effective in reducing the incidence of infectious morbidity in women undergoing cesarean section. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. The date of the most recent search was October 1998. SELECTION CRITERIA: Randomized trials that included women undergoing cesarean section were included. Trials were required to compare at least two different antibiotic regimens. Trials that compared placebo with a single antibiotic regimen were not included as these are studies which have been analyzed in another Cochrane review. DATA COLLECTION AND ANALYSIS: Data were extracted from each publication independently by the reviewers. Reviewers were not blinded to the authors or sources of the articles. The primary outcome variable was endometritis but data on other infectious complications were collected where provided. MAIN RESULTS: Fifty-one trials published between 1979 and 1994 were included in the review and four were excluded from the review. The following results refer to reductions in the incidence of endometritis. Both ampicillin and first generation cephalosporins have similar efficacy with an odds ratio (OR) of 1.27 (95% confidence interval (CI): 0.84-1.93). In comparing ampicillin with second or third generation cephalosporins the odds ratio was 0.83 (95% CI 0.54-1.26) and in comparing a first generation cephalosporin with a second or third generation agent the odds ratio was 1.21 (95% CI 0.97-1.51). A multiple dose regimen for prophylaxis appears to offer no added benefit over a single dose regimen; OR 0.92 (95% CI 0.70-1.23). Systemic and lavage routes of administration appear to have no difference in effect; OR 1.19 (95% CI 0.81-1.73). There was no significant heterogeneity between the trials contained in the various sub-group analyses, although confidence intervals were sometimes wide. REVIEWER'S CONCLUSIONS: Both ampicillin and first generation cephalosporins have similar efficacy in reducing postoperative endometritis. There does not appear to be added benefit in utilizing a more broad spectrum agent or a multiple dose regimen. There is a need for an appropriately designed randomized trial to test the optimal timing of administration (immediately after the cord is clamped versus pre-operative). PMID- 10796248 TI - Interventions for preventing and treating backache in pregnancy. AB - BACKGROUND: More than a third of women experience back pain during pregnancy. OBJECTIVES: The objective of the review was to assess the effects of preventive interventions and treatments for backache in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials of any treatment to reduce the incidence or severity of back pain in pregnancy, or to prevent back pain arising in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: One trial of 109 women was included. This cross-over trial compared the use of a special shaped pillow to fit under the woman's abdomen (Ozzlo pillow) with a standard pillow. Women experienced less pain in the week when they used the Ozzlo pillow than in the week with the standard pillow (odds ratio 0.32, 95% confidence interval 0.18 to 0.58). REVIEWER'S CONCLUSIONS: Specially shaped pillows appear to help reduce back pain in late pregnancy and improve sleep. PMID- 10796249 TI - Support for breastfeeding mothers. AB - BACKGROUND: Exclusive breastfeeding rates at three to four months remain low in many health care settings. In economically advantaged countries, young mothers, those in low-income groups or those who ceased full-time education at an early age are least likely to breastfeed. In poorer countries, more affluent groups may breastfeed less. OBJECTIVES: The objective of this review was to assess the effects of breastfeeding support. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register (last searched March 1998), Medline (from 1993), Embase (from 1980), Cinahl, Midirs (from 1991), English National Health Board database and reference lists of articles. We contacted researchers in the field. SELECTION CRITERIA: Controlled trials of acceptable quality comparing extra support for breastfeeding mothers with usual maternity care. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and checked by the second reviewer. MAIN RESULTS: Thirteen trials were included. The relative risk for stopping exclusive feeding within two months was 0. 83, 95% confidence interval 0.72 to 0.96. The relative risk for stopping breastfeeding within two months was 0.74, 95% confidence interval 0.65 to 0.86. One more mother will breastfeed for two months if support is provided for nine women (95% confidence interval 6 to 21). Similarly, one more woman will breastfeed exclusively if support is given to nine women (95% confidence interval 6 to 40). REVIEWER'S CONCLUSIONS: The provision of extra support by professionals with special skills in breastfeeding appears to result in more mothers breastfeeding their babies until two months of age, and more mothers breastfeeding their babies exclusively to two months of age. PMID- 10796250 TI - Interventions for treating constipation in pregnancy. AB - BACKGROUND: Circulating progesterone may be the cause of slower gastrointestinal movement in mid and late pregnancy. OBJECTIVES: The objective of this review was to assess the effects of different methods for treating constipation in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and Medline (1987 to 1997). SELECTION CRITERIA: Randomised trials of any treatment for constipation in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were done independently by two reviewers. MAIN RESULTS: One trial of 40 women was included. Fibre supplements increased the frequency of defaecation (odds ratio 0.18, 95% confidence interval 0.05 to 0.67), and lead to softer stools. REVIEWER'S CONCLUSIONS: Dietary supplements of fibre in the form of bran or wheat fibre are likely to help women experiencing constipation in pregnancy. PMID- 10796251 TI - Delayed (>3 weeks) postnatal corticosteroids for chronic lung disease in preterm infants. AB - BACKGROUND: Many preterm babies who survive, having had respiratory distress syndrome (RDS) or not, go on to develop chronic lung disease (CLD). This is probably due to persistence of inflammation in the lung. Corticosteroids have powerful anti-inflammatory effects and have been used to treat established CLD. However it is unclear whether any beneficial effects outweigh the adverse effects of these drugs. OBJECTIVES: To determine if late (> 3 weeks) postnatal corticosteroid treatment vs control (placebo or nothing) is of benefit in the treatment of chronic lung disease (CLD) in the preterm infant. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, the Cochrane Database of Controlled Trials, Medline, hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment initiated at > 3 weeks of age in preterm infants with CLD were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality before discharge, failure to extubate, infection, hyperglycaemia, hypertension, severe retinopathy of prematurity (ROP), necrotizing enterocolitis (NEC), gastrointestinal bleeding, bowel perforation, echodensities on ultrasound scan of brain, need for home oxygen, glycosuria, need for late rescue with dexamethasone, cerebral palsy in survivors and blindness in survivors were abstracted and analysed using Revman 4.0.4. MAIN RESULTS: Delayed steroid treatment had no effect on mortality. The only beneficial effects were reductions in failure to extubate by 7 or 28 days, need for late rescue treatment with dexamethasone, chronic lung disease at 36 weeks, and discharge to home on oxygen therapy. There was no increase in risk of infection, necrotising enterocolitis, or gastrointestinal bleeding. Short-term adverse affects included hyperglycaemia, glycosuria and hypertension. There was an increase in severe retinopathy of prematurity, but no effect on blindness. There were increases in long-term neurologic sequelae including abnormal neurologic examination and cerebral palsy, the latter of borderline statistical significance. REVIEWER'S CONCLUSIONS: The benefits of late corticosteroid therapy may not outweigh actual or potential adverse effects. There is a worrying increase in adverse neurological outcomes in infants treated with postnatal steroids (see also Review of Early Postnatal corticosteroids). Corticosteroids should be reserved for infants who cannot be weaned from mechanical ventilation. The dose of dexamethasone and the duration of any course of treatment should be minimised. PMID- 10796252 TI - Early postnatal (<96 hours) corticosteroids for preventing chronic lung disease in preterm infants. AB - BACKGROUND: Chronic lung disease (CLD) remains a major problem in neonatal intensive care units. Persistent inflammation in the lungs is the most likely underlying pathogenesis. Corticosteroids have been used to either prevent or treat CLD because of their potent anti-inflammatory effects. OBJECTIVES: To determine if postnatal corticosteroid treatment is of benefit in the prevention of chronic lung disease (CLD) in the preterm infant. This review examines the outcome of trials where preterm infants at risk of CLD were given postnatal steroids within 96 hours after birth. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, the Cochrane Controlled Trials Register, Medline, hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment within 96 hours of birth (early) in high risk preterm infants were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, failure to extubate, pulmonary air leak, survival without chronic lung disease, CLD defined at 28 days postnatal age and 36 weeks post menstrual age, patent ductus arteriosus (PDA), severe intraventricular hemorrhage (IVH), infection, hyperglycaemia, hypertension, severe retinopathy of prematurity (ROP), necrotizing enterocolitis (NEC), gastrointestinal bleeding and longterm outcome were abstracted and analysed using Revman 4.0.4. MAIN RESULTS: Fifteen randomised controlled trials of early postnatal corticosteroid treatment of preterm babies at risk of developing CLD were identified. A meta-analysis of these trials demonstrates benefits as regards earlier extubation, decreased risks of CLD at both 28 days and 36 weeks, death or CLD at 28 days, and PDA. There was an almost significant reduction in the risk of pulmonary air leak and in death or CLD at 36 weeks in the babies treated with early corticosteroids. There were no differences in the rates of neonatal mortality, infection, severe ROP, severe IVH, NEC and pulmonary haemorrhage. Gastrointestinal bleeding and intestinal perforation were important adverse effects and the risks of hyperglycaemia and hypertension were also increased. Several adverse neurological effects were found at follow-up examinations of survivors treated with early steroids: abnormal neurological examination, cerebral palsy and developmental delay. REVIEWER'S CONCLUSIONS: The benefits of early postnatal corticosteroid treatment (< 96 hours) may not outweigh the known or potential adverse effects of this treatment. Gastrointestinal effects early in the neonatal period and neurological effects seen at follow-up mean that current use of early postnatal steroids needs to be reconsidered. There is a compelling need for the long term follow-up and reporting of late outcomes, especially neurologic and developmental outcomes, among surviving infants who participated in all randomized trials of early postnatal corticosteroid treatment. The role of inhaled steroids remains to be elucidated. PMID- 10796253 TI - Furosemide for symptomatic patent ductus arteriosus in indomethacin-treated infants. AB - BACKGROUND: Inhibition of prostaglandin synthesis mediates closure of the ductus arteriosus and renal side effects after indomethacin administration. Because furosemide increases prostaglandin production, it could potentially help prevent indomethacin-related toxicity but also decrease ductal response to indomethacin. OBJECTIVES: The primary objectives of this review were to assess (1) whether furosemide affects the incidence of failure of ductal closure after indomethacin and that of indomethacin-related toxicity and (2) the effect of furosemide on mid term and long-term outcome. The secondary objective was to determine whether the effect of furosemide on renal function and water balance depends on prior extracellular volume (assessed by blood urea nitrogen [BUN]/creatinine ratio). SEARCH STRATEGY: We searched electronic databases (Medline, Embase and Cochrane) and selected abstract books, without language restriction. SELECTION CRITERIA: We selected studies with (1) random allocation to either indomethacin alone or indomethacin and furosemide and (2) analysis of either short-term risk-benefit ratio of furosemide, mid- or long-term outcome, or the relationship between extracellular volume at study entry and changes in renal function. DATA COLLECTION AND ANALYSIS: We assessed studies for possible bias and for quality of assessment of ductal patency. We assessed categorical variables using relative risk and absolute risk reduction. We assessed the effects of furosemide on renal function and fluid balance by comparing changes from baseline in the treatment group with those in controls. Subsets were determined a priori based on BUN/creatinine ratio at study entry. MAIN RESULTS: All 3 studies fulfilling the entry criteria had limitations, including possible or definite bias. There was substantial heterogeneity among studies. Furosemide administration did not significantly increase the risk of failure of ductal closure; however, sample size was insufficient to rule out even a 31% increase. In the subset with initial BUN/creatinine ratio > 20 mg/mg, 2 of 18 patients receiving furosemide could not complete a 3-dose course of indomethacin because of toxicity. Minimal or no information was available about any of the other main outcome variables. Furosemide increased urine output regardless of the initial BUN/creatinine ratio, leading to a 5% weight loss during a 3-dose course, an undesired effect in patients with initial BUN/creatinine ratio > 20 mg/mg. Furosemide increased creatinine clearance only in patients with initial BUN/creatinine ratio <20 mg/mg. REVIEWER'S CONCLUSIONS: There is not enough evidence to support the administration of furosemide to premature infants treated with indomethacin for symptomatic patent ductus arteriosus. Furosemide appears to be contraindicated in the presence of dehydration in those infants. PMID- 10796254 TI - Emollient ointment for preventing infection in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effect of prophylactic application of emollient ointment in preterm infants. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: ointment; limits: age groups, newborn infant; publication types, clinical trial), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, and journal handsearching in the English language. SELECTION CRITERIA: Randomized controlled trials which compared the effect of prophylactic application of emollient ointment to routine care or as needed topical therapy in preterm infants are included in this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including transepidermal water loss, skin condition, fluid intake, suspect infection and proven nosocomial infection were excerpted from the reports of the clinical trials by the reviewers. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Two randomized trials which compared prophylactic application of ointment to routine skin care or as needed topical ointment therapy were identified. Lane (1993) noted improved skin condition in infants receiving topical application of emollient ointment. In the study of Nopper and coworkers (1996), prophylactic application of ointment significantly decreased transepidermal water loss during the first six hours after initial application. Skin condition was noted to be improved during the first 1-2 weeks. Surveillance cultures demonstrated less bacterial colonization during the two week study. A significant decrease in suspect and proven infection was noted. Fewer infants were evaluated for sepsis among the group who received prophylactic application of ointment (relative risk 0. 50, 95% CI 0.27, 0.93; risk difference -0.30, 95% CI -0.54, -0.06). Both studies reported on the incidence of proven nosocomial infection. A trend towards a decrease in the risk of proven nosocomial infection was noted in infants who received prophylactic application of emollient ointment (typical relative risk 0.29, 95% CI 0.07, 1.16, typical risk difference -0.13, 95% CI -0.25, -0.01). REVIEWER'S CONCLUSIONS: In two small studies, prophylactic application of emollient ointment decreased transepidermal water loss, decreased the severity of dermatitis, and decreased the risk of suspect sepsis and proven sepsis. Further clinical studies are warranted to validate these results. PMID- 10796255 TI - Types of intra-muscular opioids for maternal pain relief in labour. AB - BACKGROUND: Pethidine is the most widely used intra-muscular opioid for the relief of labour pain. However concerns have been raised about its effectiveness and the possibility of depressing respiration in newborns. OBJECTIVES: The objective of this review was to assess the effects of different opioids (and different doses of the same opioid) administered intra-muscularly in labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register (Cochrane Library, issue 4, 1997) and reference lists of articles. SELECTION CRITERIA: Randomised trials comparing the effects of different currently used opioids (and different doses of the same opioid) administered intramuscularly in labour for women who request systemic analgesia. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. Analysis was based on the groups as randomly allocated. MAIN RESULTS: Sixteen trials were included. There were problems with methodological quality of some of the trials, and lack of consistency in the way various outcomes were reported. There was no evidence of a difference between pethidine and tramadol in terms of pain relief, interval to delivery, or instrumental or operative delivery. There appeared to be more adverse effects such as nausea and vomiting and drowsiness with pethidine. Maternal pain relief seemed almost identical between the meptazinol and pethidine groups, whether assessed as maternal satisfaction with pain relief, visual analogue scales, or use of other pain relief, but meptazinol gave rise to slightly more side effects. Maternal satisfaction with pain relief appeared similar for pentazocine and pethidine, with more frequent nausea and vomiting with pethidine. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the comparative efficacy and safety of the various opioids used for analgesia in labour. PMID- 10796256 TI - Intravenous immunoglobulin for suspected or subsequently proven infection in neonates. AB - BACKGROUND: Congenital and nosocomial infections are important causes of neonatal morbidity and mortality. Maternal transport of immunoglobulins to the fetus mainly occurs after 32 weeks gestation and endogenous synthesis does not begin until several months after birth. Administration of intravenous immunoglobulin provides IgG that can bind to cell surface receptors, provide opsonic activity, activate complement, promote antibody dependent cytotoxicity, and improve neutrophilic chemoluminescence. Theoretically infectious morbidity and morbidity could be reduced by the administration of intravenous immunoglobulin. OBJECTIVES: To assess the effectiveness of intravenous immunoglobulin (IVIG) to reduce mortality/morbidity caused by suspected infection in newborn infants. In secondary analyses to assess the effectiveness of IVIG to reduce mortality/morbidity in those neonates who entered into the studies with suspected infection and who later were confirmed as being infected. SEARCH STRATEGY: Medline, Embase, and Reference Update Databases were searched in November 1997, and the Cochrane Library in July 1998 using the following keywords: immunoglobulin and infant-newborn, and random allocation, or controlled trial, or randomized controlled trial (RCT). The reference lists of identified RCTs and meta-analyses, personal files and Science Citation Index were also searched. No language restrictions were applied. Unpublished data were requested from authors and information has been obtained from one author to date. SELECTION CRITERIA: The criteria used to select studies for inclusion were: 1) DESIGN: RCT (including quasi-randomized trials) 2) Newborn infants (< 28 days old) 3) INTERVENTION: IVIG for treatment of suspected (and in some infants subsequently proved) bacterial/fungal infection compared to placebo or no intervention. Suspected infection was defined as clinical symptoms and signs consistent with infection without isolation of causative organism. Proved infection was defined as: clinical symptoms and signs consistent with infection in association with isolation of causative organism from either blood culture, cerebrospinal fluid culture, urine culture (urine obtained by suprapubic tap) or a normally sterile site (e.g., liver, spleen, meninges, lung) at autopsy. 4) At least one of the following outcomes was reported: mortality during initial hospital stay; length of hospital stay; side effects; psychomotor development/growth at follow up. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted information for the outcomes of interest and one researcher (AO) checked for any discrepancies and pooled the results. Relative risk (RR) and Risk Difference (RD) with 95% confidence intervals (CI) using the fixed effects model are reported for dichotomous outcomes and weighted mean difference (WMD) for continuous data. NNT was calculated for outcomes that showed a statistically significant reduction in RD. Data from quasi-randomized trials were excluded in sensitivity analyses. MAIN RESULTS: Study quality was generally poor. Four of 7 identified studies (n = 208), reported on the outcomes of all randomized patients with clinically suspected infection. Mortality was reduced [RR 0.52 (95% CI; 0.28, 0.98), RD 0.102 (95% CI; -0.005, -0.199, NNT 10 (95% CI; 5, 200]. When, in a sensitivity analysis (n = 126), the results from a quasi-randomized trial (n = 82) were excluded, RR and RD remained similar, [RR 0.53 (95% CI; 0.25, 1.15), RD -0.106 ( 0.232, 0.021)] but statistical significance was lost. Treatment with IVIG (six trials, n = 234) in cases of subsequently proved infection did not result in a statistically significant reduction in mortality [RR 0.62 (95% CI; 0.34, 1.12, RD -0.074 (95% CI; -0.163, 0. 014)]. Excluding in a sensitivity analysis (n = 199) a quasi-randomized trial (n = 35) changed the results slightly [RR 0. 68 (95% CI; 0.36, 1.29), RD -0.059 (-0.152, 0.035)]. There was no statistically significant between-study PMID- 10796257 TI - Anticonvulsants for preventing mortality and morbidity in full term newborns with perinatal asphyxia. AB - OBJECTIVES: To assess the benefits and harm of administering anticonvulsants to infants of 37 weeks gestation or more following perinatal asphyxia with the primary aims of prevention of death or subsequent severe neurodevelopmental disability and/or the prevention of seizures. SEARCH STRATEGY: Relevant randomised controlled trials were identified using a combination of electronic database searches (MEDLINE), hand searches and a search of the Neonatal Review Group trials register. SELECTION CRITERIA: All randomised, or quasi-randomised, controlled clinical trials with reported data comparing the following outcomes: mortality, neurodevelopmental disability, neonatal seizures and adverse events, following anticonvulsant therapy in term infants (37 weeks or more), compared to controls with or without placebo, following perinatal asphyxia. DATA COLLECTION AND ANALYSIS: Methodological quality and validity of studies were assessed without consideration of the results. Data relevant to the outcome were extracted and analysed. MAIN RESULTS: Five randomised or quasi-randomised controlled trials which met the selection criteria were identified. No studies were of sufficient methodological quality and size to demonstrate a valid, clinically significant change in the risk of mortality or severe neurodevelopmental disability. A meta analysis combining three studies comparing barbiturates with conventional therapy following perinatal asphyxia demonstrated no difference in risks of death, severe neurodevelopmental disability, or death or severe neurodevelopmental disability. REVIEWER'S CONCLUSIONS: At the present time, anticonvulsant therapy to term infants in the immediate period following perinatal asphyxia cannot be recommended for routine clinical practice, other than in the treatment of prolonged or frequent clinical seizures. Any future studies should be of high quality: randomised control trials with allocation concealment, performance and outcome assessment blinding. Such studies should be of sufficient size, with minimal attrition, to have the power to detect clinically important reductions in mortality and severe neurodevelopmental disability, as the primary outcome measures. PMID- 10796259 TI - Umbilical vein injection for management of retained placenta. PMID- 10796258 TI - Dopamine versus dobutamine for hypotensive preterm infants. AB - BACKGROUND: Inotropes are widely used in preterm infants to treat systemic hypotension. The most commonly used drugs are dopamine and dobutamine. These agents have different modes of action which may result in different haemodynamic effects. OBJECTIVES: To compare the effectiveness and safety of dopamine and dobutamine in the treatment of systemic hypotension in preterm infants. SEARCH STRATEGY: The standard search method of the Cochrane Neonatal Review Group was used. Searches of electronic and other databases were performed. Previous reviews were searched for references to relevant trials and leading authors in the field were contacted for information about other published and unpublished studies. SELECTION CRITERIA: Randomised controlled trials where short and/or long term effects of treatment with dopamine and dobutamine for the treatment of systemic arterial hypotension were compared were selected for this review. Trials studying newborn infants born before 37 completed weeks gestation and less than 28 days of age were eligible for inclusion. Systemic arterial hypotension was not defined specifically, but accepted as defined in individual studies. Studies were not limited by birthweight, lower gestational age threshold or by route or duration of administration of inotropic agents. Study quality and eligibility were assessed independently by each reviewer. DATA COLLECTION AND ANALYSIS: The standard method of the Cochrane Collaboration described in the Cochrane Collaboration Handbook was used to perform this systematic review. Data extraction was performed independently by each reviewer, with differences being resolved by discussion. The following outcomes were determined: mortality in the neonatal period, long term neurodevelopmental outcome, radiological evidence of severe neurological injury, short term haemodynamic changes and incidence of adverse effects. The effect of interventions is expressed either as Relative Risk (RR), Risk Difference (RD) or as Weighted Mean Difference (WMD) with their 95% Confidence Interval (CI). MAIN RESULTS: Four trials met the pre-defined criteria for inclusion in this review. There was no evidence of a significant difference between dopamine and dobutamine in terms of neonatal mortality (RD 0.02 95% CI 0.12 to 0.16), incidence of periventricular leukomalacia (RD -0.08, 95% CI -0.19 to 0.04), or severe periventricular haemorrhage (RD -0.02, 95% CI -0.13 to 0.09). Dopamine was more successful than dobutamine in treating systemic hypotension, with fewer infants having treatment failure (RD -0.29, 95% CI -0.42 to -0.17; NNT = 3.5, 95% CI 2.4 to 5.9). There was no evidence of a significant difference in change in left ventricular output when dopamine was compared with dobutamine (WMD -83 ml/kg/min, 95% CI -174 to 8 ml/kg/min). There was no evidence of a significant difference between the two agents with respect to the incidence of tachycardia (RD -0.06, 95% CI -0.25 to 0.14). None of the studies reported the incidence of adverse long term neurodevelopmental outcome. REVIEWER'S CONCLUSIONS: Dopamine is more effective than dobutamine in the short term treatment of systemic hypotension in preterm infants. There was no evidence of an effect on the incidence of adverse neuroradiological sequelae (severe periventricular haemorrhage and/or periventricular leucomalacia), or on the incidence of tachycardia. However, in the absence of data confirming long term benefit and safety of dopamine compared to dobutamine, no firm recommendations can be made regarding the choice of drug to treat hypotension. PMID- 10796260 TI - Oral misoprostol for induction of labour with a viable fetus. AB - BACKGROUND: Misoprostol is a synthetic prostaglandin which has been used to induce labour. Oral use of the drug misoprostol may be convenient, but an overdose could cause uterine hyperstimulation and precipitate labour which may be life-threatening for both mother and fetus. OBJECTIVES: The objective of this review was to assess the effects of oral misoprostol used for labour induction in women with a viable fetus. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials of oral misoprostol versus any other method, placebo or no treatment given to women with a viable fetus for induction of labour. DATA COLLECTION AND ANALYSIS: The selection of trials and data extraction were undertaken by one reviewer and independently checked by two other reviewers. MAIN RESULTS: Five trials were included. In one placebo trial, oral misoprostol reduced the need for oxytocin infusion and shortened delivery time in women with prelabour rupture of membranes at term. In another trial, compared to vaginal prostaglandins, oral misoprostol reduced the need for oxytocin (relative risk 0.62, 95% confidence interval 0.47 to 0.82). Based on two trials, the caesarean section rate with oral misoprostol was 20. 2% (55/272) compared with 15.5% (42/270) for vaginal prostaglandins (relative risk 1.29, 95% confidence interval 0.90 to 1.86). Different doses (100 micrograms three hourly and 200 micrograms six hourly) were used in the two trials that compared oral with vaginal misoprostol. The caesarean section rate was 21.8% in the oral misoprostol group compared with 13.5% for vaginal misoprostol (relative risk 1.62, 95% confidence interval 0.85 to 3.09). The uterine hyperstimulation rate with oral misoprostol was 37.5% (36/96) compared with 28% (25/89) for vaginal misoprostol (relative risk 1.32, 95% confidence interval 0.86 to 2.04). There was significant heterogeneity between these two trials. REVIEWER'S CONCLUSIONS: Oral misoprostol may be an effective method for labour induction. However clinically effective oral regimens may have an unacceptably high incidence of uterine hyperstimulation and possibly uterine rupture. PMID- 10796261 TI - Drugs for rapid treatment of very high blood pressure during pregnancy. AB - BACKGROUND: Very high blood pressure during pregnancy poses a serious threat to women and their fetuses. The use of drugs to lower blood pressure may reduce this risk. OBJECTIVES: The objective of this review was to compare different antihypertensive drugs used for rapid treatment of severe hypertension during pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: STUDIES: All randomised trials. Quasi random designs were excluded. PARTICIPANTS: Women with severe hypertension during pregnancy. Women postpartum at trial entry were excluded. INTERVENTIONS: Comparisons of one antihypertensive agent with another. OUTCOMES: For the women: blood pressure control, eclampsia, serious maternal morbidity (such as kidney failure and liver failure), Caesarean section, and use of health service resources (such as admission to hospital or intensive care unit). For the baby: death, serious neonatal morbidity, infant and child development, and use of health service resources (such as admission to a special care nursery). DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers to assess eligibility and describe the trial characteristics, and by one reviewer for the meta-analyses. Discrepancies were resolved by discussion. There was no blinding of authorship or results. Whenever possible, unpublished data were sought from investigators. MAIN RESULTS: Thirteen of the 14 trials included in this review were small (range 19-627 women). Of the eight comparisons, five included hydralazine. Diazoxide given as 75mg bolus injections appears to be associated with profound hypotension requiring treatment, and ketanserin is less effective than hydralazine at reducing blood pressure. There is no other evidence that any one of the other antihypertensive agents is better than another for women with severe hypertension during pregnancy. REVIEWER'S CONCLUSIONS: Until better evidence is available, the choice of antihypertensive should depend on the experience and familiarity of an individual clinician with a particular drug, and on what is known about adverse maternal and fetal side-effects. Exceptions are diazoxide and ketanserin, which are probably not good choices. PMID- 10796262 TI - Routine doppler ultrasound in pregnancy. AB - BACKGROUND: Doppler ultrasound study of umbilical artery waveforms helps identify the compromised fetus in 'high risk' pregnancies and, therefore, deserves assessment as a screening test in 'low risk' pregnancies. One of the main aims of routine antenatal care is to identify the 'at risk' fetus in order to apply clinical interventions which could result in reduced perinatal morbidity and mortality. OBJECTIVES: To assess the effects on obstetric practice and pregnancy outcome of routine Doppler ultrasound in unselected and low risk pregnancies. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group Specialised Register of Controlled Trials and the Cochrane Controlled Trials Register were searched. Date of last search: September 1999 SELECTION CRITERIA: Acceptably controlled trials of routine Doppler ultrasound (umbilical circulation and/or uterine circulation) in unselected or low risk pregnancies. DATA COLLECTION AND ANALYSIS: Both reviewers assessed trial quality and extracted data. Authors of two trials were contacted for additional information. MAIN RESULTS: Five trials were included which recruited 14,338 women. The methodological quality of the trials was generally good. Based on existing evidence, routine Doppler ultrasound examination in low risk or unselected populations did not result in increased antenatal, obstetric and neonatal interventions, and no overall differences were detected for substantive short term clinical outcomes such as perinatal mortality. There is no available evidence to assess the effect on substantive long term outcomes such as childhood neurodevelopment. There is no available evidence to assess maternal outcomes, particularly psychological effects. In two studies there were unexpected findings suggesting possible harmful effects, but the explanation for this is not clear, and further evaluation regarding the safety of Doppler ultrasound is required. REVIEWER'S CONCLUSIONS: Based on existing evidence, routine Doppler ultrasound in low risk or unselected populations does not confer benefit on mother or baby. Future research should be powerful enough to address small changes in perinatal outcome, and should include evaluation of maternal psychological effects, long term outcomes such as neurodevelopment, and issues of safety. PMID- 10796263 TI - Routine ultrasound in late pregnancy (after 24 weeks gestation). AB - BACKGROUND: Diagnostic ultrasound is used selectively in late pregnancy where there are specific clinical indications. However, the value of routine late pregnancy ultrasound screening in unselected populations is controversial. The rationale for such screening would be the detection of clinical conditions which place the fetus or mother at high risk, which would not necessarily have been detected by other means such as clinical examination, and for which subsequent management would improve perinatal outcome. OBJECTIVES: To assess the effects on obstetric practice and pregnancy outcome of routine late pregnancy ultrasound, defined as greater than 24 weeks gestation, in women with either unselected or low risk pregnancies. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group Specialised Register of Controlled Trials and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: All acceptably controlled trials of routine ultrasound in late pregnancy (defined as after 24 weeks). DATA COLLECTION AND ANALYSIS: The principal reviewer assessed trial quality and extracted data, under supervision of the co-reviewer. MAIN RESULTS: Seven trials recruiting 25,036 women were included. The quality of trials overall was satisfactory. There was no difference in antenatal, obstetric and neonatal intervention or morbidity in screened versus control groups. Routine late pregnancy ultrasound was not associated with improvements in overall perinatal mortality. Placental grading as an adjunct to third trimester examination scan was associated with a significant reduction in the stillbirth rate in the one trial that assessed it. There is a lack of data with regard to long term substantive outcomes such as neurodevelopment. There is a lack of data on maternal psychological effects. REVIEWER'S CONCLUSIONS: Based on existing evidence, routine late pregnancy ultrasound in low risk or unselected populations does not confer benefit on mother or baby. There is a lack of data about the potential psychological effects of routine ultrasound in late pregnancy, and the effects on both short and long term neonatal and childhood outcome. Placental grading in the third trimester may be valuable, but whether reported results are reproducible remains to be seen, and future research of late pregnancy ultrasound should include evaluation of placental textural assessment. PMID- 10796264 TI - Intravenous or enteral loop diuretics for preterm infants with (or developing) chronic lung disease. AB - BACKGROUND: Lung disease in preterm infants is often complicated with lung edema. OBJECTIVES: The aim of this review was to assess the risks and benefits of administration of a diuretic acting on the loop of Henle (loop diuretic) in preterm infants with or developing chronic lung disease (CLD). Primary objectives were to assess changes in need for oxygen or ventilatory support and effects on long-term outcome, and secondary objectives were to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We used the following keywords: ? or ? and , limited to and limited to or . We searched Medline (1966-1998), Embase (1974-1998) and the Cochrane Controlled Trials Register (CCTR) from the Cochrane Library (1998, issue 4). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing chronic lung disease and at least 5 days of age were all randomly allocated to receive a loop diuretic either enterally or intravenously. Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included important clinical outcomes, and secondary outcome variables included toxicity and pulmonary mechanics (e.g., lung compliance and airway resistance). DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: The only loop diuretic used in the studies which met the selection criteria was furosemide. Most studies focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review, or the potential complications of diuretic therapy. In preterm infants < 3 weeks of age developing CLD, furosemide administration has either inconsistent effects or no detectable effect. In infants > 3 weeks of age with CLD, a single intravenous dose of 1 mg/kg of furosemide improves lung compliance and airway resistance for 1 hour. Chronic administration of furosemide improves both oxygenation and lung compliance. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks of age with CLD, acute and chronic administration of furosemide improve lung compliance. Chronic administration of intravenous or enteral furosemide improves oxygenation. In view of the lack of data from randomized trials concerning effects on important clinical outcomes, routine or sustained use of systemic loop diuretics in infants with (or developing) CLD cannot be recommended based on current evidence. Randomized trials are needed to assess the effects of furosemide administration on survival, duration of ventilatory support and oxygen administration, length of hospital stay, potential complications and long-term outcome. PMID- 10796265 TI - Diuretics for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Lung edema may complicate respiratory distress syndrome (RDS) in preterm infants. OBJECTIVES: The aim of this review was to assess the risks and benefits of diuretic administration in preterm infants with RDS. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We searched Medline, Embase and the Cochrane Controlled Trials Register from the Cochrane Library, using the following keywords: and . In addition, we searched the abstract books of the American Thoracic Society and Pediatric Research Societies. SELECTION CRITERIA: We only included trials in which preterm infants with RDS and less than 5 days of age were randomly allocated to diuretic administration. Of those trials, we only included studies in which at least one of the following outcomes measures was evaluated: mortality, patent ductus arteriosus, hypovolemic shock, intraventricular hemorrhage, renal failure, duration of oxygen supplementation, duration of mechanical ventilation, need for oxygen supplementation at 28 days of life, oxygen supplementation at 36 weeks of postconceptional age (gestational age + postnatal age), length of stay, number of rehospitalizations during the first year of life, and neurodevelopmental outcome. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study. Any disagreement was resolved by discussion. MAIN RESULTS: Six studies met inclusion criteria. Studies available for this systematic review were all done before the current era of prenatal steroids, surfactant, indomethacin and fluid restriction. Furosemide administration had no long-term benefits. Furosemide-induced transient improvement in pulmonary function did not outweigh an increased risk for patent ductus arteriosus and for hemodynamic instability. REVIEWER'S CONCLUSIONS: There are no current data to support routine diuretic administration in preterm infants with RDS. Elective administration of furosemide or any diuretic to any patient with RDS should be carefully weighed against the risk of precipitating hypovolemia. In addition, elective administration of furosemide should be weighed against the risk of developing a symptomatic patent ductus arteriosus. PMID- 10796266 TI - Early versus delayed selective surfactant treatment for neonatal respiratory distress syndrome. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To compare the effects of early vs. delayed selective surfactant therapy for newborns intubated for respiratory distress within the first two hours of life. Planned subgroup analyses include separate comparisons for studies utilizing natural surfactant extract and synthetic surfactant. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactant; text word: early; limits: age, newborn: publication type, clinical trial), PubMed, abstracts, conference and symposia proceedings, expert informants, and journal hand searching in the English language. SELECTION CRITERIA: Only randomized controlled clinical trials comparing early selective surfactant administration (surfactant administration via the endotracheal tube in infants intubated for respiratory distress, not specifically for surfactant dosage) within the first 2 hours of life versus delayed selective surfactant administration to infants with established respiratory distress syndrome were considered for review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including the incidence of pneumothorax, patent ductus arteriosus, pulmonary interstitial emphysema, pulmonary hemorrhage, necrotizing enterocolitis, retinopathy of prematurity, intraventricular hemorrhage (any and severe IVH), bronchopulmonary dysplasia, chronic lung disease, neonatal mortality, mortality prior to hospital discharge, bronchopulmonary dysplasia or death, and chronic lung disease or death were excerpted from the reports of the clinical trials by the reviewers. Data regarding the average number of surfactant doses per infant were also analyzed. Further analysis of data with regard to surfactant type was performed. Data analysis was performed in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Four randomized controlled trials met selection criteria. Two of the trials utilized synthetic surfactant (Exosurf Neonatal) and two utilized a natural surfactant extract. The meta-analyses demonstrated significant reductions in risk of pneumothorax (Typical RR 0.70, 95%CI 0.59, 0.82; Typical RD -0.05, 95%CI -0.08, -0.03), and pulmonary interstitial emphysema (Typical RR 0.63, 95%CI 0.43, 0.93; Typical RD -0.06, 95%CI -0.10, -0.01) in infants randomized to early selective surfactant administration. Infants randomized to early selective surfactant administration also demonstrated a decreased risk of neonatal mortality (Typical RR 0.87, 95%CI 0.77, 0.99; Typical RD -0.03, 95%CI -0.06, -0.00), chronic lung disease (Typical RR 0.70, 95%CI 0. 55, 0.88; Typical RD -0.03, 95%CI -0.05, -0.01), and chronic lung disease or death at 36 weeks (Typical RR 0.84, 95%CI 0.75, 0.93; Typical RD -0.06, 95%CI 0.09, -0.03). A trend toward risk reduction for bronchopulmonary dysplasia or death at 28 days was also evident (Typical RR 0.94, 95%CI 0.88, 1.00; Typical RD 0.04, 95%CI -0.07, -0.00). No differences in other complications of RDS or prematurity were noted. REVIEWER'S CONCLUSIONS: Early selective surfactant administration given to infants with RDS requiring assisted ventilation leads to a decreased risk of acute pulmonary injury (decreased risk of pneumothorax and pulmonary interstitial emphysema) and a decreased risk of neonatal mortality and chronic lung disease compared to delaying treatment of such infants until they develop established RDS. PMID- 10796267 TI - Glutamine supplementation for preventing morbidity in preterm infants. AB - BACKGROUND: The amino acid glutamine is the preferred respiratory fuel for rapidly proliferating cells under normal conditions. Recent research has suggested a number of roles for glutamine during critical illness. This research has been largely performed in experimental animals and in adults in a variety of disease settings. There is little information on the role of glutamine in children and infants, or whether glutamine supplementation is beneficial in preterm babies. OBJECTIVES: To determine the effects of glutamine supplementation on morbidity and weight gain in preterm babies. SEARCH STRATEGY: Searches were made using Medline and Embase electronic databases and specific handsearching in the English language. The search strategy followed the guidelines of the Neonatal Cochrane Review Group. SELECTION CRITERIA: Randomised controlled trials comparing glutamine supplementation to no glutamine supplementation in preterm babies at any time from birth to discharge from hospital. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including duration of parenteral nutrition, time to full enteral nutrition, rate of weight gain, rate of positive blood cultures and duration of hospital stay were extracted by both reviewers. Analysis was performed by the primary reviewer (TRJT) in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Three trials met the selection criteria. Data on proportion of babies having one or more of positive blood cultures were available from all three studies. Meta-analysis showed no significant difference between glutamine-supplemented and non-supplemented babies; RR = 0.73 (95% CI 0.44, 1.23), RD = -8.8% (95% CI -23.2, 5.5). Data for other outcome variables were pooled from two studies. There were no significant differences between glutamine-supplemented and non-supplemented babies for days to full enteral nutrition (WMD 0.42, 95% CI -3.0, 3.8), rate of weight gain (WMD 0.6 g/kg/d, 95% CI -1.6, 2.8) or days of hospital stay (WMD -2.4, 95% CI -14.9, 10.2). REVIEWER'S CONCLUSIONS: There is no evidence to support the routine use of parenteral or enteral glutamine supplementation in preterm babies. A large randomised controlled trial should be performed to determine whether or not glutamine supplementation enhances gut integrity and reduces sepsis rate. PMID- 10796268 TI - Treatments for toxoplasmosis in pregnancy. AB - BACKGROUND: Toxoplasmosis is a widespread parasitic disease and usually causes no symptoms. However, infection of pregnant women may cause congenital infection, resulting potentially in mental retardation and blindness in the infant. OBJECTIVES: The objective of this review was to assess whether or not treating toxoplasmosis in pregnancy reduces the risk of congenital toxoplasma infection and improves infant outcomes. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. An electronic search was performed using the key words 'congenital and toxoplasmosis' on the following databases: MEDLINE (1966-07/1997), Embase (1993-07/1997), Pascal (French) (1990-1997), Biological Abstracts (1993-1995) and the Cochrane Controlled Trials Register. There was also contact with experts in the field, including those in the European Research Network on Congenital Toxoplasmosis. SELECTION CRITERIA: Randomised controlled trials of antibiotic treatment versus no treatment of pregnant women with proven or likely acute Toxoplasma infection, with outcomes in the children reported. We also inspected relevant reports of less robust experimental studies in which there were (non randomly allocated) control groups, although it was not planned to include such data in the primary analysis. DATA COLLECTION AND ANALYSIS: Reports of possibly eligible studies were scrutinised by two investigators. MAIN RESULTS: Out of the 2591 papers identified, none met the inclusion criteria. REVIEWER'S CONCLUSIONS: Despite the large number of studies performed over the last three decades we still do not know whether antenatal treatment in women with presumed toxoplasmosis reduces the congenital transmission of Toxoplasma gondii. Screening is expensive, so we need to evaluate the effects of treatment, and the impact of screening programmes. In countries where screening or treatment is not routine, these technologies should not be introduced outside the context of a carefully controlled trial. PMID- 10796269 TI - Reduced salt intake compared to normal dietary salt, or high intake, in pregnancy. AB - BACKGROUND: In the past women have been advised that lowering their salt intake might reduce their risk of pre-eclampsia. Although this practice has largely ceased, it remains important to assess the evidence about possible effects of advice to alter dietary salt intake during pregnancy. OBJECTIVES: The objective of this review was to assess the effects of dietary advice to alter salt intake compared to continuing a normal diet, on the risk of pre-eclampsia and its consequences. SEARCH STRATEGY: We searched the register of trials maintained and updated by the Cochrane Pregnancy and Childbirth Group, and the Cochrane Controlled Trials Register Disc Issue 4, 1998. SELECTION CRITERIA: Studies were included if they were randomised trials of advice to either reduce or to increase dietary salt during pregnancy. DATA COLLECTION AND ANALYSIS: All data were extracted independently by both reviewers. MAIN RESULTS: Two trials were included, with 603 women. They compared advice about a low salt diet with no dietary advice. The confidence intervals for all of the outcomes reported were wide, and cross the no effect line. This includes pre-eclampsia (relative risk 1.11, 95% confidence interval 0.46 to 2.66). Even when taken together, these trials are insufficient to provide reliable information about the effects of advice on salt restriction during normal pregnancy. None of the trials included women with pre-eclampsia, so this review provides no reliable information about changes in salt intake for treatment of pre-eclampsia. REVIEWER'S CONCLUSIONS: Salt consumption during pregnancy should remain a matter of personal preference. PMID- 10796270 TI - Oestrogens and progestogens for preventing and treating postnatal depression. AB - BACKGROUND: Postnatal depression, with a prevalence of at least 10%, is probably the most common complication of the puerperium. A deficiency or imbalance of sex hormones has repeatedly been suggested as a cause. OBJECTIVES: The objective of this review was to evaluate the role of oestrogens and progestogens in the prevention and treatment of postnatal depression. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. SELECTION CRITERIA: All trials were considered in which pregnant or postpartum women (up to 18 months) were randomised to receive postpartum oestrogen or progestogen or placebo for the treatment or prevention of postnatal depression. DATA COLLECTION AND ANALYSIS: Two published randomised placebo controlled trials were identified for inclusion in the analyses for this review. One study was excluded. MAIN RESULTS: Depot norethisterone enanthate given within 48 hours of delivery and lasting 8-12 weeks was associated with significantly higher postpartum depression scores than placebo. Oestrogen therapy in severely depressed women was associated with a greater improvement in depression scores than placebo. REVIEWER'S CONCLUSIONS: There is no place for synthetic progestogens in the prevention of treatment of postnatal depression. Long-acting norethisterone enanthate is associated with an increased risk of postnatal depression. It and other long-acting progestogen contraceptives should be used with caution in the postnatal period, especially in women with a history of depression. The role of progesterone in the prevention and treatment of postnatal depression has yet to be evaluated in a randomised placebo-controlled trial. Oestrogen therapy may be of modest value at a late stage of severe postnatal depression. Its role in the prevention of recurrent postnatal depression has not been evaluated. Further research on its value is unlikely for ethical reasons. PMID- 10796271 TI - Aerosolized diuretics for preterm infants with (or developing) chronic lung disease. AB - BACKGROUND: Lung disease in preterm infants is often complicated with lung edema. OBJECTIVES: The aim of this review is to assess the risks and benefits of aerosolized diuretic administration in preterm infants with or developing chronic lung disease (CLD). Primary objectives are to assess effects on short term outcome (changes in need for oxygen or ventilatory support) and effects on long term outcome. Secondary objectives are to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We used the following keywords: ? or ? and , limited to and limited to or . We searched Medline (1966-1998), Embase (1974-1998) and the Cochrane Controlled Trials Register (CCTR) from the Cochrane Library (1998, Issue 4). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing chronic lung disease and at least five days of age were all randomly allocated to receive an aerosolized loop diuretic. Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included important clinical outcomes, and secondary outcome variables included pulmonary mechanics and potential complications of therapy. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: We identified eight studies which met selection criteria. Most studies focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review or the potential complications of diuretic therapy. No study assessed the amount of diuretic effectively delivered to the patient. Furosemide was the only diuretic used in the eight studies included in this review. Among preterm infants < 3 weeks of age developing CLD, not enough information is available to assess the effect of aerosolized furosemide on outcome or lung function. Among infants > 3 weeks with CLD, a single aerosolized dose of 1 mg/kg of furosemide may transiently improve pulmonary mechanics. Not enough information is available to assess the effect of chronic administration of aerosolized furosemide on oxygenation and pulmonary mechanics. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks with CLD administration of a single dose of aerosolized furosemide improves pulmonary mechanics. In view of the lack of data from randomized trials concerning effects on important clinical outcomes, routine or sustained use of aerosolized loop diuretics in infants with (or developing) CLD cannot be recommended based on current evidence. More double-blinded randomized trials are needed (1) to analyze factors likely to affect the response to aerosolized furosemide, e.g. , washout period and delivery of furosemide to distal airways, and (2) to assess the effects of chronic administration of aerosolized furosemide on mortality, O2 dependency, ventilator dependency, length of hospital stay and long-term outcome. PMID- 10796272 TI - Plasma volume expansion for treatment of women with pre-eclampsia. AB - BACKGROUND: Plasma volume is reduced amongst women with pre-eclampsia. This association has led to the suggestion that expanding the plasma volume might improve maternal and uteroplacental circulation, and so potentially improve outcome for both the woman and her baby. OBJECTIVES: The aim of this review was to assess the effects of plasma volume expansion for the treatment of women with pre-eclampsia. SEARCH STRATEGY: The register of trials maintained by the Cochrane Pregnancy and Childbirth Group, and the Cochrane Controlled Trials Register Issue 1 1999 were searched for trials meeting the selection criteria. SELECTION CRITERIA: Randomised trials were included. Quasi-random designs were excluded. Participants were women with hypertension during pregnancy, with or without proteinuria. Women who were postpartum at trial entry were excluded. Interventions were any comparison of plasma volume expansion with no expansion, or of one plasma volume expander with another. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Discrepancies were resolved by discussion. There was no blinding of authorship or results. MAIN RESULTS: Three trials involving 61 women were included in this review. All compared a colloid solution with no plasma volume expansion. For every outcome reported, the confidence intervals are very wide and cross the no effect line. REVIEWER'S CONCLUSIONS: There is insufficient evidence for any reliable estimates of the effects of plasma volume expansion for women with pre-eclampsia. PMID- 10796273 TI - Erythromycin for feeding intolerance in preterm infants. AB - BACKGROUND: Functional immaturity of gastointestinal motility predisposes preterm infants to feeding intolerance. Motilin, a gastrointestinal peptide, stimulates propagative contractile activity during phase III of the migratory motor complex in the interdigestive state. Erythromycin (EM) is a motilin agonist with prokinetic effect at low doses (1-3mg/kg). OBJECTIVES: To evaluate the effectiveness of EM in promoting gastrointestinal motility in preterm infants with feeding intolerance and assess clinically significant adverse effects associated with its use. SEARCH STRATEGY: Systematic literature search in accordance with the Cochrane Neonatal Collaborative Review Group search strategy. Randomized and quasi-randomized controlled trials of EM use, at any dose, in preterm infants to promote gastrointestinal motility were identified by searching MEDLINE, EMBASE, CINAHL, the Cochrane Library, reference lists of published studies, personal files, and abstracts published in Pediatric Research. SELECTION CRITERIA: Randomized controlled trials of oral or intravenous EM use at dose range of 3 to 12 mg/kg/day in preterm infants less than or equal to 36 weeks gestational age with feeding tolerance were included in this review. DATA COLLECTION AND ANALYSIS: Data regarding the primary clinical outcome of days to achieve full enteral feeding were compared among studies. Data on secondary outcomes including adverse effects associated with the use of EM (diarrhea, nosocomial infections, cardiac arrhythmias, or theophylline toxicity), duration of parenteral nutrition, weight gain, incidence of necrotizing enterocolitis (NEC), hypertrophic pyloric stenosis, and length of hospital stay were assessed. MAIN RESULTS: Two randomized controlled studies of EM use in preterm infants for improving gastrointestinal motility were identified. Since both studies involved preterm infants treated with EM at dose >12mg/kg/day at commencement of feeding, they did not meet inclusion criteria defined a priori for this review. There was no statistically significant difference in the incidence of NEC (RR 0.59, 95%CI 0.11, 3.01; RD -0.021, 95%CI -0. 087, 0.045). No statistically significant difference was noted in days to achieve full enteral feeds, length of hospital stay, and adverse events between groups. REVIEWER'S CONCLUSIONS: EM at antimicrobial doses may not be effective in preterm infants with feeding intolerance. Further studies are needed to determine whether EM in lower doses is effective as a prokinetic agent in such infants. PMID- 10796274 TI - Diuretics acting on the distal renal tubule for preterm infants with (or developing) chronic lung disease. AB - OBJECTIVES: The aim of this review is to assess the risks and benefits of diuretics acting on distal segments of the renal tubule (distal diuretics) in preterm infants with or developing chronic lung disease (CLD). Primary objectives are to assess changes in need for oxygen or ventilatory support and effects on long-term outcome, and secondary objectives are to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard method of the Cochrane Neonatal Review Group. We used the following keywords: ? or ? and , limited to and limited to or . We searched Medline (1966-1998), Embase (1974-1998) and the Cochrane Controlled Trials Register (CCTR) from the Cochrane Library (1999, issue 2). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing CLD and at least five days of age were all randomly allocated to receive a distal diuretic (i.e., a diuretic acting on the distal renal tubule). Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included changes in need for respiratory support and oxygen supplementation, mortality, bronchopulmonary dysplasia (BPD), death or BPD, chronic lung disease at 36 weeks of postconceptional age (gestational age + postnatal age), length of stay, and number of rehospitalizations during the first year of life. Secondary outcome variables included pulmonary mechanics and potential complications of therapy. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: Of six studies fulfilling entry criteria, most focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review, or the potential complications of diuretic therapy. In preterm infants > 3 weeks of age with CLD, a four-week treatment with thiazide and spironolactone improved lung compliance and reduced the need for furosemide. Thiazide and spironolactone decreased the risk of death and tended to decrease the risk for lack of extubation after 8 weeks in intubated infants. There is no evidence to support the hypothesis that adding spironolactone to thiazide or that adding metolazone to furosemide improves the outcome of preterm infants with CLD. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks of age with CLD, acute and chronic administration of distal diuretics improve pulmonary mechanics. Chronic administration of diuretics (thiazide with spironolactone) reduces mortality in intubated patients. Large studies are needed to assess (1) whether chronic diuretic administration improves mortality, duration of oxygen dependency, ventilator dependency, length of hospital stay and long-term outcome in patients without long-term toxicity and (2) whether adding spironolactone to thiazides or adding metolazone to furosemide has any beneficial effect. PMID- 10796275 TI - Early administration of inhaled corticosteroids for preventing chronic lung disease in ventilated very low birth weight preterm neonates. AB - BACKGROUND: Chronic lung disease remains a common complication amongst preterm infants. There is increasing evidence that inflammation play an important role in the pathogenesis of CLD. Due to their strong anti-inflammatory properties corticosteroids is an attractive intervention strategy. However, there are growing concerns regarding short and long term effects of systemic corticosteroids. Theoretically, administration of inhaled corticosteroids may allow for beneficial effects on the pulmonary system with a lower risk of undesirable systemic side effects. OBJECTIVES: To determine the impact of inhaled corticosteroids administered to ventilated very low birth weight preterm neonates in the first two weeks of life for the prevention of chronic lung disease(CLD). SEARCH STRATEGY: Systematic search in accordance with Cochrane Neonatal Review Group. Randomized and quasi-randomized trials were identified by searching MEDLINE, Embase, CINAHL, the Cochrane Library, reference lists of published trials and abstracts published in Pediatric Research. SELECTION CRITERIA: Randomized controlled trials of inhaled corticosteroid therapy initiated within the first 2 weeks of life in ventilated preterm infants with birth weight 1500 grams or less were included in this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including chronic lung disease at 28 days or 36 weeks corrected gestational age (CGA), mortality, combined outcome of death or CLD at 28 days of age and at 36 weeks CGA, the need for systemic corticosteroids, failure to extubate within 14 days and adverse effects of corticosteroids were evaluated. All data were analyzed using Revman 3.1. When possible, meta-analysis was performed using relative risk (RR), risk difference (RD), along with their 95% confidence intervals (CI). If RD was significant, number needed to treat (NNT) was calculated. MAIN RESULTS: Eight trials assessing the impact of inhaled corticosteroid for the prevention of CLD were identified. The study by Kovacs 1998 was excluded as investigators evaluated the impact of a combination of systemic and inhaled corticosteroid for prevention of CLD. Seven trials qualified for inclusion in this review but data from two of these studies are awaiting assessment. Thus, the present review includes data analyses based on five qualifying trials. There was no statistically significant effect of inhaled steroids on CLD either at 28 days or at 36 weeks CGA, when analyzed either for all randomized infants or amongst survivors. No statistically significant differences were noted for mortality or for the combined outcome of mortality and CLD either at 28 days of age or at 36 weeks CGA. The meta-analysis supports a reduction in the need for systemic steroids, RR 0.78 (95% CI 0.62, 0.99), RD -0. 097 (95% CI -0.187, -0.008); however statistical heterogeneity was noted. The number needed to treat (NNT) to reduce the need for systemic steroid was 10 (95% CI 5.3, 125). There were no statistically significant differences in adverse events between groups. REVIEWER'S CONCLUSIONS: There is no evidence from the trials reviewed that early administration (in the first 2 weeks of life) of inhaled steroids to ventilated preterm neonates was effective in reducing the incidence of CLD. There was a reduction in the need for systemic steroids. Although this difference was statistically significant, there was significant heterogeneity between studies and the upper limit of the 95% CI for this outcome was very close to no effect. Currently, use of inhaled steroids in this population cannot be recommended. Studies are needed to identify the risk/benefit ratio of different delivery techniques and dosing schedules for the administration of these medications. Studies need to address both the short-term and long-term benefits and adverse effects of inhaled steroids with particular attention to neurodevelopmental outcome. PMID- 10796276 TI - Early versus delayed initiation of progressive enteral feedings for parenterally fed low birth weight or preterm infants. AB - BACKGROUND: Enteral feedings in very-low-birth-weight or sick preterm infants are often delayed for several days or weeks after birth even though delayed enteral feeding could diminish the functional adaptation of the gastrointestinal tract and result in feeding intolerance later. Early initiation of feedings, if well tolerated, may promote growth and shorten the duration of parenteral nutrition and hospital stay without increasing the risk for necrotizing enterocolitis (NEC). OBJECTIVES: For parenterally fed low-birth-weight infants, to assess the effects of early enteral feedings initiated shortly after birth compared to delayed enteral feedings (with similar schedules for advancing feedings in each group). SEARCH STRATEGY: Searches were performed of the Oxford Database of Perinatal Trials, the Cochrane Neonatal Review Group registry, MEDLINE, abstracts and conference proceedings, references from relevant publications in the English language, and studies identified by personal communication. SELECTION CRITERIA: Only randomized or quasi-randomized clinical trials were considered. Trials were included if 1) they enrolled low birth weight or preterm infants who were all given parenteral nutrition; 2) the infants were randomly assigned to either early enteral feedings (mean or median age <=4 days) or late enteral feedings (>4 days) of formula or breast milk; 3) except when feeding intolerance developed, the feedings were progressively advanced starting within 72 hours after initiating feedings; and 4) the goals for total nutrient intake were similar for both groups. (We did not require the duration or total intake of parenteral nutrients to be similar for both groups because these variables may be affected by the age at which feedings are initiated.) DATA COLLECTION AND ANALYSIS: The two reviewers reached consensus for inclusion of trials. Data regarding clinical outcomes were extracted and evaluated by two reviewers (JET and KAK) independently. Authors were contacted as needed and feasible to clarify or provide missing data. The specific data that were needed were requested in writing and by telephone. MAIN RESULTS: Only two small studies were identified (one with 60 patients and one with 12 patients). Five randomized trials were excluded because parenteral nutrition was not provided or because the groups were assigned to receive different parenteral intakes as well as different enteral intakes. An additional unpublished small trial was excluded because both groups were fed "late" according to our categorization. Because there were no clinical outcomes which were reported in both of the included studies, no meta analysis of the results was performed. Based on the results of the individual studies, early feedings had no significant effect on weight gain, necrotizing enterocolitis, mortality, or age at discharge, although important effects cannot be excluded with the small number of patients studied. Some benefits of early feedings were noted in the larger trial (Davey) -- fewer days on parenteral nutrition, fewer infants who were treated with gastric suction and interruption of feedings, fewer infants with sepsis evaluations, and fewer infants with percutaneous central venous catheters. REVIEWER'S CONCLUSIONS: The benefits and hazards of early and delayed feedings have received very little study in clinical trials, and the effects on major clinical outcomes, including necrotizing enterocolitis and death, remain uncertain. With the availability of parenteral nutrition in contemporary neonatal units, it is unclear whether high-risk infants should receive early or delayed feedings. PMID- 10796277 TI - Elective delivery in diabetic pregnant women. AB - BACKGROUND: In pregnancies complicated by diabetes the major concerns during the third trimester are fetal distress and the potential for birth trauma associated with fetal macrosomia. OBJECTIVES: The objective of this review was to assess the effect of a policy of elective delivery, as compared to expectant management, in term diabetic pregnant women, on maternal and perinatal mortality and morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (last searched July 1999). SELECTION CRITERIA: All available randomized controlled trials of elective delivery, either by induction of labour or by elective caesarean section, compared to expectant management in diabetic pregnant women at term. DATA COLLECTION AND ANALYSIS: The reports of the only available trial were analysed independently by the three co-reviewers to retrieve data on maternal and perinatal outcomes. Results are expressed as relative risks (RR) and 95% confidence intervals (CI). MAIN RESULTS: The participants in the one trial included in this review were 200 insulin-requiring diabetic women. Most had gestational diabetes, except 13 women with type 2 preexisting diabetes (class B). The trial compared a policy of active induction of labour at 38 completed weeks of pregnancy, to expectant management until 42 weeks. The risk of caesarean section was not statistically different between groups (RR 0.81, 95% CI 0.52 - 1.26). The risk of macrosomia was reduced in the active induction group (RR 0.56, 95%CI 0.32 - 0. 98) and 3 cases of mild shoulder dystocia were reported in the expectant management group. No other perinatal morbidity was reported. REVIEWER'S CONCLUSIONS: There is very little evidence to support either elective delivery or expectant management at term in pregnant women with insulin-requiring diabetes. Limited data from a single randomized controlled trial suggest that induction of labour in women with gestational diabetes treated with insulin reduces the risk of macrosomia. Although the small sample size does not permit one to draw conclusions, the risk of maternal or neonatal morbidity was not modified. Women's views on elective delivery and on prolonged surveillance and treatment with insulin should be assessed in future trials. PMID- 10796278 TI - Interventions for suspected placenta praevia. AB - BACKGROUND: Because placenta praevia is implanted unusually low in the uterus, it may cause major, and/or repeated, antepartum haemorrhage. The traditional policy of care of women with symptomatic placenta praevia includes prolonged stay in hospital and delivery by caesarean section. OBJECTIVES: To assess the impact of any clinical intervention applied specifically because of a perceived likelihood that a pregnant woman might have placenta praevia. SEARCH STRATEGY: A comprehensive electronic search was performed to identify relevant literature. Searched databases included the Trials Register maintained by the Cochrane Pregnancy and Childbirth Group, and the Cochrane Controlled Clinical Trials Register. SELECTION CRITERIA: Any controlled clinical trial that has assessed the impact of an intervention in women diagnosed as having, or being likely to have, placenta praevia. DATA COLLECTION AND ANALYSIS: Data were extracted from the three identified trial reports, unblinded, by the author without consideration of results. MAIN RESULTS: Two comparisons could be made - home versus hospitalisation and cervical cerclage versus no cerclage. Both were associated with reduced lengths of stay in hospital antenatally. Otherwise, there was little evidence of any clear advantage or disadvantage to a policy of home versus hospital care. Cervical cerclage may reduce the risk of delivery before 34 weeks, or the birth of a baby weighing less than 2 kg or having a low 5 minute Apgar score. In general, these possible benefits were more evident in the trial of lesser methodological quality. REVIEWER'S CONCLUSIONS: There are insufficient data from trials to recommend any change in clinical practice. Available data should, however, should encourage further work to address the safety of more conservative policies of hospitalisation for women with suspected placenta praevia, and the possible value of insertion of a cervical suture. PMID- 10796279 TI - Woman's position during second stage of labour. AB - BACKGROUND: For centuries, there has been controversy around whether being upright (sitting, birthing stools, chairs, squatting) or lying down have advantages for women delivering their babies. OBJECTIVES: The objective of this review was to assess the benefits and risks of the use of different positions during the second stage of labour (i. e. from full dilatation of the cervix). SEARCH STRATEGY: Relevant trials are identified from the register of trials maintained by the Cochrane Pregnancy and Childbirth Group, and from the Cochrane Controlled Trials Register. SELECTION CRITERIA: Trials were included which compared various positions assumed by pregnant women during the second stage of labour. Randomised and quasi-randomised trials with appropriate follow-up were included. DATA COLLECTION AND ANALYSIS: Trials were independently assessed for inclusion, and data extracted, by the two authors. Disagreements would have been resolved by consensus with an editor. Meta-analysis of data is performed using the RevMan software. MAIN RESULTS: Results should be interpreted with caution as the methodological quality of the 18 trials was variable. Use of any upright or lateral position, compared with supine or lithotomy positions, was associated with: 1. Reduced duration of second stage of labour (12 trials - mean 5.4 minutes, 95% confidence interval (CI) 3.9 - 6.9 minutes). This was largely due to a considerable reduction in women allocated to use of the birth cushion. 2. A small reduction in assisted deliveries (17 trials - odds ratio (OR) 0.82, 95% CI 0.69 - 0.98). 3. A reduction in episiotomies (11 trials - OR 0.73, 95% CI 0.64 - 0.84). 4. A smaller increase in second degree perineal tears (10 trials - OR 1.30, 95% CI 1.09 - 1.54). 5. Increased estimated risk of blood loss > 500ml (10 trials - OR 1.76, 95% CI 1.34 - 3.32). 6. Reduced reporting of severe pain during second stage of labour (1 trial - OR 0.59, 95% CI 0.41 - 0.83). 7. Fewer abnormal fetal heart rate patterns (1 trial - OR 0.31, 95% CI 0.11 - 0.91). REVIEWER'S CONCLUSIONS: The tentative findings of this review suggest several possible benefits for upright posture, with the possibility of increased risk of blood loss > 500ml. Women should be encouraged to give birth in the position they find most comfortable. Until such time the benefits and risks of various delivery positions are estimated with greater certainty when methodologically stringent trials data are available, then women should be allowed to make informed choices about the birth positions in which they might wish to assume for delivery of their babies. PMID- 10796280 TI - Intravenous midazolam infusion for sedation of infants in the neonatal intensive care unit. AB - BACKGROUND: The need for sedation for neonates undergoing uncomfortable procedures in the neonatal intensive care unit (NICU) has often been overlooked. Proper sedation may reduce stress and avoid complications during procedures such as mechanical ventilation. Midazolam is a short acting benzodiazepine that has been increasingly used in the NICU. However, the effectiveness of intravenous midazolam as a sedative in neonates has not been systematically evaluated. OBJECTIVES: To determine whether intravenous midazolam infusion is an effective sedative, as evaluated by behavioural and/or physiologic measurements, for critically ill neonates undergoing intensive care, and to assess clinically significant short and long term adverse effects associated with its use. SEARCH STRATEGY: Literature search according to the Cochrane Neonatal Collaborative Review Group search strategy. Randomized and quasi-randomized controlled trials of intravenous midazolam use in neonates were identified by searching MEDLINE, EMBASE, the Cochrane Controlled Trials Register, reference lists of published studies, personal files, and abstracts published in Pediatric Research from 1990 1999. SELECTION CRITERIA: Randomized controlled trials of intravenous midazolam infusion in infants = 5d) or no enteral nutrient intake (no feedings or water only). DATA COLLECTION AND ANALYSIS: The two reviewers reached consensus for inclusion of trials. Data regarding clinical outcomes were extracted and evaluated by the two reviewers independently of each other. Authors were contacted as needed and feasible to clarify or provide missing data. The specific data that were needed were requested in writing. MAIN RESULTS: Among infants given minimal enteral nutrition (MEN), there was an overall reduction in days to full enteral feeding, total days that feedings were held, and total hospital stay. There was no discernible effect on necrotizing enterocolitis. REVIEWER'S CONCLUSIONS: The evidence of benefit from MEN in these analyses is not convincing for a variety of reasons--the inherent difficulty of assessing enteral feedings in high-risk infants, the small size and methodologic limitations of the studies to date, unexplained heterogeneity with respect to some of the apparent benefits, the potential for bias to affect the findings in unblinded studies, and the unexcluded possibility that MEN might increase necrotizing enterocolitis. For these reasons, it is unclear whether MEN should be used in lieu of an equal period of time without enteral feedings. PMID- 10796376 TI - Nebulized racemic epinephrine for extubation of newborn infants. AB - BACKGROUND: Following a period of mechanical ventilation, post-extubation upper airway obstruction can occur in newborn infants, especially after prolonged, traumatic or multiple intubations. The subsequent increase in upper airway resistance may lead to respiratory insufficiency and failure of extubation. The vasoconstrictive properties of epinephrine, and its proven efficacy in the treatment of croup in infants, has led to the routine use of inhaled nebulised epinephrine immediately post-extubation in some neonatal units. It is also recommended for neonates with post-extubation tracheal obstruction and stridor in neonatal and respiratory textbooks and reviews. OBJECTIVES: The primary objective was to assess whether nebulised epinephrine administered immediately after extubation in neonates weaned from IPPV decreases the need for subsequent additional respiratory support. SEARCH STRATEGY: Searches were made of Medline (MeSH search terms 'epinephrine' and 'exp infant, newborn'), the Oxford Database of Perinatal trials, expert informants and journal hand searching mainly in the English language, expert informant searches in the Japanese language by Prof. Ogawa, previous reviews including cross references, abstracts, and conference and symposia proceedings. SELECTION CRITERIA: All randomised and quasi-randomised control trials in which nebulised epinephrine was compared with placebo immediately post-extubation in newborn infants who have been weaned from IPPV and extubated, with regard to clinically important outcomes (i.e. need for additional respiratory support, increase in oxygen requirement, respiratory distress, stridor or the occurrence of side effects). DATA COLLECTION AND ANALYSIS: No studies met our criteria for inclusion in this review. MAIN RESULTS: No studies were identified which looked at the effect of inhaled nebulised epinephrine on clinically important outcomes in infants being extubated. REVIEWER'S CONCLUSIONS: IMPLICATIONS FOR PRACTICE: There is no evidence either supporting or refuting the use of inhaled nebulised racemic epinephrine in newborn infants. IMPLICATIONS FOR RESEARCH: randomised controlled trials are needed comparing inhaled nebulised racemic epinephrine with placebo in neonates post-extubation. This should be looked at both as a routine treatment post-extubation and as specific treatment for post-extubation upper airway obstruction. Study populations should include the group of infants at highest risk for upper airway obstruction from mucosal swelling because of their small glottic and sub-glottic diameters (ie those infants with birthweights less than 1000 grams). PMID- 10796377 TI - Umbilical artery catheters in the newborn: effects of heparin. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine whether the use of heparin in fluids infused through an umbilical arterial catheter in newborn infants influences the frequency of clinical ischemic events, catheter occlusion, aortic thrombosis, intraventricular hemorrhage, hypertension, death, or the duration of catheter usability. SEARCH STRATEGY: Randomized and quasi randomized controlled trials of umbilical catheterization use were obtained from the following sources: 1. Effective Care of the Newborn Infant, edited by JC Sinclair and MB Bracken. 2. Medline Search using Melvyl Medline Plus and the keyword headings 'Umbilic#', 'Catheter#' and subject heading 'Infant, Newborn' 3. Search of personal data files SELECTION CRITERIA: Randomized trials in newborn infants of any birthweight or gestation. Comparison of heparinised to non heparinised infusion fluids, including comparison of heparin in the infusate to heparin just in the flush solution. Clinically important end points such as catheter occlusion or aortic thrombosis. DATA COLLECTION AND ANALYSIS: There were five randomized controlled trials retrieved. All gave details of the incidence of catheter occlusion. Two also reported the incidence of aortic thrombosis. The intervention was reasonably consistent: heparin in the infusate at a concentration of 1 unit/mL was investigated in all trials except one which used a concentration of 0.25 units/mL. Studies generally included both term and preterm infants. MAIN RESULTS: Heparinization of the infusate decreases the incidence of catheter occlusion but does not affect the frequency of aortic thrombosis. Heparinization of the flush solution is not an adequate alternative. There does not appear to be an effect on frequency of intraventricular hemorrhage, death or clinical ischemic phenomena. REVIEWER'S CONCLUSIONS: Heparinization of the fluid infused through an umbilical arterial catheter decreases the likelihood of umbilical arterial catheters occluding. The lowest concentration tested so far (0.25 units/mL) has been shown to be effective. Heparinization of flushes without heparinizing the infusate is ineffective. The frequency of aortic thrombosis has not been shown to be affected; however, the confidence intervals for this effect are very wide. The frequency of intraventricular hemorrhage has not been shown to be affected by heparinization of the infusate, but again the confidence intervals are very wide and even a major increase in the incidence of grade 3 and 4 intraventricular hemorrhage would not have been detected. PMID- 10796378 TI - Umbilical artery catheters in the newborn: effects of catheter design (end vs side hole). AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine whether the design of an umbilical arterial catheter influences the frequency of ischemic events, aortic thrombosis, intraventricular hemorrhage, mortality or necrotising enterocolitis in newborn infants. SEARCH STRATEGY: Randomized and quasi randomized controlled trials of umbilical catheterization use were obtained from the following sources: 1. Effective Care of the Newborn Infant, edited by JC Sinclair and MB Bracken. 2. Medline Search using Melvyl Medline Plus and the keyword headings 'Umbilic#', 'Catheter#' and subject heading 'Infant, Newborn' 3. Search of personal data files SELECTION CRITERIA: Randomized trials in newborn infants of any birthweight or gestation. Comparison of end hole catheters with side hole catheters. Clinically important end points such as ischemic events or aortic thrombosis. DATA COLLECTION AND ANALYSIS: There appears to be only a single trial which has addressed this issue (Wesstrom 1979). MAIN RESULTS: End hole catheters are associated with a much decreased risk of aortic thrombosis compared to side hole catheters. RR = 0.27 (95% CI 0.11, 0.67) REVIEWER'S CONCLUSIONS: Side hole catheters should be avoided for umbilical arterial catheterisation in the newborn. PMID- 10796379 TI - Prophylactic versus selective use of surfactant for preventing morbidity and mortality in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To compare the effect of prophylactic surfactant administration to surfactant treatment of established respiratory distress syndrome in premature infants. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactant; limits: age groups, newborn infants), previous reviews including cross-references, abstracts, conference and symposia proceedings, expert informants, and journal handsearching in the English language. SELECTION CRITERIA: Randomized controlled trials which compared the effects of prophylactic surfactant administration to surfactant treatment of established respiratory distress syndrome in premature infants were included in the analysis. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including the incidence of pneumothorax, pulmonary interstitial emphysema, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage (any grade and severe intraventricular hemorrhage), bronchopulmonary dysplasia, mortality, bronchopulmonary dysplasia or death, and retinopathy of prematurity were excerpted from the reports of the clinical trials by the reviewers. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: The majority of included studies noted an initial improvement in the respiratory status and a decrease in the incidence of respiratory distress syndrome in infants who received prophylactic surfactant. The meta-analysis supports a decrease in the incidence of pneumothorax, a decrease in the incidence of pulmonary interstitial emphysema, a decrease in the incidence of mortality and a decrease in the incidence of bronchopulmonary dysplasia or death associated with prophylactic administration of surfactant. No significant untoward effects of prophylactic surfactant administration are noted. REVIEWER'S CONCLUSIONS: Prophylactic surfactant administration to infants judged to be at risk of developing respiratory distress syndrome (intubated infants less than 30-32 weeks gestation) has been demonstrated to improve clinical outcome. Infants who receive prophylactic surfactant have a decreased incidence of pneumothorax, a decreased incidence of pulmonary interstitial emphysema and a decreased incidence of mortality. However, it remains unclear exactly which criteria should be used to judge "at risk" infants who would require prophylactic surfactant administration. PMID- 10796380 TI - Prophylactic natural surfactant extract for preventing morbidity and mortality in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effect of prophylactic intratracheal administration of natural surfactant extract in preterm newborns at risk for developing respiratory distress syndrome (RDS). SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactant; limits: age groups; newborn infants), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants and journal hand searching in the English language. SELECTION CRITERIA: Randomized controlled trials which compared the effect of prophylactic natural surfactant administration (surfactant obtained from human or bovine sources, either modified with additional phospholipids or not) administered to high risk preterm newborns at or shortly after birth in order to prevent respiratory distress syndrome, other complications of prematurity, and mortality. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including incidence of pneumothorax, pulmonary interstitial emphysema, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage (any grade and severe intraventricular hemorrhage), bronchopulmonary dysplasia, mortality, bronchopulmonary dysplasia or death, and retinopathy of prematurity were excerpted from the reports of the clinical trials by the reviewer. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: All of the included studies note an initial improvement in respiratory status and a decrease in the risk of respiratory distress syndrome in infants who receive prophylactic natural surfactant extract. The meta-analysis supports a decrease in the risk of pneumothorax (typical relative risk 0.35, 95% CI 0.26, 0.49; typical risk difference -0.15, 95% CI -0.20, -0.11), a decrease in the risk pulmonary interstitial emphysema (typical relative risk 0.46, 95% CI 0.35, 0.60; typical risk difference -0.19, 95% CI -0.25, -0.13), a decrease in the risk of neonatal mortality (typical relative risk 0. 60, 95% CI 0.44, 0.83; typical risk difference -0.07, 95% CI -0.12, -0.03), and a decrease in the risk of bronchopulmonary dysplasia or death (typical relative risk 0.84, 95% CI 0.75, 0.93; typical risk difference -0.10, 95% CI -0.16, -0.04. No differences are reported in the risk of intraventricular hemorrhage, patent ductus arteriosus, necrotizing enterocolitis or retinopathy of prematurity. Few data are available on long-term followup of treated infants. REVIEWER'S CONCLUSIONS: Prophylactic intratracheal administration of natural surfactant extract to infants judged to be at risk of developing respiratory distress syndrome (intubated infants <30 weeks gestation) has been demonstrated to improve clinical outcome. Infants who receive prophylactic natural surfactant extract have a decreased risk of pneumothorax, a decreased risk of pulmonary interstitial emphysema, a decreased risk of mortality, and a decreased risk of bronchopulmonary dysplasia or death. PMID- 10796381 TI - Thrombolysis (different doses, routes of administration and agents) for acute ischaemic stroke. AB - BACKGROUND: Thrombolytic therapy is effective for acute myocardial infarction, a vascular disease with some similarities to acute ischaemic stroke. OBJECTIVES: The objective of this review was to assess the effect of different thrombolytic agents, and different regimens in acute ischaemic stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register and Embase (1980 to 1997). We handsearched Japanese and Chinese journals. We contacted researchers in the field and pharmaceutical companies. SELECTION CRITERIA: Randomised and quasi-randomised trials of different doses of a thrombolytic agent, or one thrombolytic agent compared with another, or the same agent given by different routes, in people with confirmed acute ischaemic stroke. Trials were included if treatment was started within 14 days of stroke onset. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility, trial quality and extracted the data. MAIN RESULTS: Eight trials involving 1334 people were included. Concealment of allocation was generally adequate. All the trials were conducted in Japan. Different doses (of tissue plasminogen activator or urokinase) were compared in seven trials. Different agents (tissue plasminogen activator versus urokinase, or tissue-cultured urokinase versus conventional urokinase) were compared in three trials. Few data were available for functional outcomes. A higher dose of thrombolytic therapy was associated with a five-fold increase in fatal intracranial haemorrhages (odds ratio 5.02, 95% confidence interval 1.56 to 16. 18). This was based on 11 events among 369 higher-dose patients and one event among 356 lower-dose patients in six trials. There was a non-significant trend towards more early deaths or clinically significant intracranial haemorrhages. No difference in late deaths or extra-cranial haemorrhages was shown between low and higher doses. However, very few of these events occurred. No difference was shown between the different thrombolytic agents tested. REVIEWER'S CONCLUSIONS: There is not enough evidence to conclude whether lower doses of thrombolytic agents might be safer or more effective than higher doses in acute ischaemic stroke. It is not possible to conclude whether one agent might be better than another, or which route of administration might be best. No comparative data for streptokinase have been found. PMID- 10796382 TI - Percutaneous transluminal angioplasty and stenting for carotid artery stenosis. AB - BACKGROUND: Carotid percutaneous transluminal angioplasty may be a useful non surgical procedure for carotid stenosis, particularly for lesions not suitable for surgery. OBJECTIVES: The objective of this review was to assess the effect of carotid percutaneous transluminal angioplasty compared with carotid endarterectomy (in patients suitable for surgery) or medical therapy (in patients not suitable for surgery). SEARCH STRATEGY: We searched the Cochrane Stroke Groups trials register (to August 1997). We contacted researchers in the field and balloon catheter manufacturers. SELECTION CRITERIA: Randomised trials of carotid percutaneous transluminal angioplasty (with or without stenting) compared with carotid endarterectomy, or percutaneous transluminal angioplasty plus best medical therapy compared with best medical therapy alone, in people with carotid artery stenosis (symptomatic or asymptomatic). DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the inclusion criteria. MAIN RESULTS: No completed trials were found. Two trials are underway. REVIEWER'S CONCLUSIONS: There is no evidence as yet to assess the relative effects of carotid percutaneous transluminal angioplasty in people with carotid stenosis. PMID- 10796383 TI - Percutaneous transluminal angioplasty and stenting for vertebral artery stenosis. AB - BACKGROUND: Surgery to the vertebrobasilar region is difficult and not considered in most centres. Furthermore, the basilar artery, with its penetrating branches supplying the brain stem, is likely to be extremely hazardous to dilate. OBJECTIVES: The objective of this review was to assess the effect of vertebral artery percutaneous transluminal angioplasty combined with medical care, compared to medical care alone, in people with vertebrobasilar artery disease. SEARCH STRATEGY: We searched the Cochrane Stroke Groups trials register (to August 1997). We contacted researchers in the field and balloon catheter manufacturers. SELECTION CRITERIA: Randomised trials of vertebral percutaneous transluminal angioplasty (with or without stenting) combined with best medical therapy, compared with best medical therapy alone, in people with symptomatic vertebral artery stenosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the inclusion criteria. MAIN RESULTS: No completed trials were found. One trial is underway, although few patients with vertebrobasilar disease have been recruited. REVIEWER'S CONCLUSIONS: There is no evidence as yet to assess the effects of percutaneous transluminal angioplasty for vertebral artery stenosis. PMID- 10796384 TI - Analgesia and non-aspirin, non-steroidal anti-inflammatory drugs for osteoarthritis of the hip. AB - OBJECTIVES: To review all randomized trials of analgesics and anti-inflammatory therapy in osteoarthritis (OA) of the hip. To determine which non-steroidal, anti inflammatory drug (NSAID) is the most effective, and which NSAID is the most toxic. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register and Medline up to August 1994. Reference lists of all trials were also manually searched. SELECTION CRITERIA: All randomized controlled trials comparing non-steroidal anti-inflammatory drugs (NSAIDs) and/or analgesics in patients with Osteoarthritis. The trials selected for inclusion were identified by one reviewer (TT) and rechecked by a second (MH). DATA COLLECTION AND ANALYSIS: Qualitative assessments were performed using a quality scoring system designed for NSAID trials in rheumatoid arthritis. Both the design and analysis aspects of the trials were evaluated, each aspect being rated on a scale of 0 to 8. A quantitative method, which calculates the ratio of improvement produced by one NSAID to that produced by another, was used to rate the relative efficacy of different NSAIDs with respect to pain relief. Toxicity comparisons were made according to the reviewer findings. All quality assessments were carried out independently by two reviewers (TT, BS). All data abstraction was carried out by one reviewer (TT) and rechecked by two other reviewers (BS, GW). A consensus was reached on discrepancies. MAIN RESULTS: Forty-three trials were identified, and of these, 39 evaluated NSAIDs, while four evaluated only analgesics. The median design and analysis scores were two and four respectively. Six NSAIDs were included in at least five trials. Of these, indomethacin was rated more effective in five of its seven comparisons, but more toxic in seven of 12 comparisons. Only five of the 29 (17%) NSAID comparisons found statistically significant differences in efficacy. Of the 43 RCTs identified only 17 had statistical data available for future pooling for this meta-analysis. In the case where data was missing, authors of the trials will be contacted for inclusion of data in future reviews. REVIEWER'S CONCLUSIONS: NSAID trials in patients with OA of the hip appear to be weakened by the lack of standardization of case definition of OA, and also by the lack of standardization of outcome assessments. No clear recommendations for the choice of specific NSAID therapy in hip OA can be offered at this time based on this analysis. PMID- 10796385 TI - Balneotherapy for rheumatoid arthritis and osteoarthritis. AB - BACKGROUND: Balneotherapy (hydrotherapy or spa therapy) for patients with arthritis is one of the oldest forms of therapy. One of the aims of balneotherapy is to soothe the pain and as a consequence to relieve patients' suffering and make them feel well. OBJECTIVES: To perform a systematic review to assess the effects of balneotherapy for rheumatoid arthritis and osteoarthritis. SEARCH STRATEGY: Using the Cochrane search strategy, studies were found by screening: 1) The Medline CD-ROM database from 1966 to June 1999 and 2) the database from the Cochrane Field 'Rehabilitation and Related Therapies', which contains also studies published in journals not covered by Medline. Also, 3) reference checking and 4) personal communications with authors was carried out to retrieve eligible studies. To perform an adequate assessment of the methodological quality the languages of the publications had to be: Dutch, English, French or German. Date of the most recent literature search: June, 1999 SELECTION CRITERIA: Studies were eligible if they were randomized controlled trials (RCT) comparing balneotherapy with any intervention or with no intervention. Patients included had rheumatoid arthritis (RA), osteoarthritis (OA) or some other form of arthritis. Trials incorporating patients with definite or classical rheumatoid arthritis (RA) as defined by the American Rheumatism Association Criteria (ARA) (Ropes 1958) (these criteria have changed over time) or by the criteria of Steinbrocker (1949) were regarded as a separate group. At least one of the WHO/ILAR core set of endpoints for RA clinical trials had to be the main outcome measures. DATA COLLECTION AND ANALYSIS: A criteria list used to assess the methodological quality was the one developed at the Department of Epidemiology at the Maastricht University, called "the Maastricht list". The quality scores and data abstraction of the studies were carried out independently by two reviewers (HdV, RdB). Disagreements were solved by consensus. MAIN RESULTS: Ten trials with 607 patients were included in this review. Most trials reported positive findings, but were methodologically flawed to some extent. A 'quality of life' outcome was reported by two trials. Just one of the randomized trials mentioned an intention-to-treat analysis and only three performed a comparison of effects between groups. Pooling of the data was not performed, because of heterogeneity of the studies, multiple outcome measurements, and, apart from two studies, the overall data presentation was too scarce to enable pooling of the data. REVIEWER'S CONCLUSIONS: One cannot ignore the positive findings reported in most trials. However the scientific evidence is weak because of the poor methodological quality, the absence of an adequate statistical analysis, and the absence, for the patient, of most essential outcome measures (pain, quality of life), Therefore, the noted "positive findings" should be viewed with caution. Because of the methodological flaws an answer about the efficacy of balneotherapy cannot be provided at this time. Flaws found in the reviewed studies could be avoided in future trials. PMID- 10796386 TI - Injectable gold for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term benefit and risk of side-effects of injectable gold for rheumatoid arthritis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, and Medline, up to July 1997, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. SELECTION CRITERIA: Randomized clinical trials (RCT) comparing injectable gold against placebo in patients with rheumatoid arthritis were included. DATA COLLECTION AND ANALYSIS: Methodological quality of the RCTs was asessed by two reviewers (MS, BS) (kappa=1.0). Rheumatoid arthritis outcome measures were extracted by two reviewers from the publications for the 6 month endpoint. Sufficient data was obtained to conduct a pooled analysis of the number of swollen joints, physician global assessment, patient global assessment and erythrocyte sedimentation rate (ESR). Results were analyzed as standardized weighted mean differences for swollen joints and global assessments and weighted mean differences for ESR. Toxicity was evaluated with pooled odds ratios for withdrawals. Heterogeneity was estimated using a chi-square test. Fixed effects models were used throughout. MAIN RESULTS: Four trials and 415 patients were included. A statistically significant benefit was observed for injectable gold when compared to placebo. The standardized weighted difference (effect size) between gold and placebo for the number of swollen joints was -0.5, translating into a percentage change of 30% in favour of gold adjusted for placebo. Statistically significant differences were also observed for ESR and patient and physician assessments. Twenty two percent of the treated patients withdrew from toxicity compared to 4% of controls (OR=3.9 - 95%Cl: 2.1 - 7.2). REVIEWER'S CONCLUSIONS: Although its use can be limited by the incidence of serious toxicity, injectable gold has an important clinically and statistically significant benefit in the short term treatment of patients with rheumatoid arthritis. PMID- 10796387 TI - General versus spinal/epidural anaesthesia for surgery for hip fractures in adults. AB - BACKGROUND: The majority of hip fracture patients are treated surgically, requiring anaesthesia. OBJECTIVES: To compare different types of anaesthesia for surgical repair of hip fractures (proximal femoral fractures) in adults. This is primarily regional (spinal or epidural) anaesthesia versus inhalation general anaesthesia, but also includes ketamine anaesthesia versus inhalation general anaesthesia. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, Medline, selected orthopaedic and anaesthetic journals and conference proceedings, and reference lists of relevant articles. Date of the most recent search: August 1998. SELECTION CRITERIA: Randomised and quasi randomised trials comparing different methods of anaesthesia for hip fracture surgery in skeletally mature persons. Trials comparing the use of local nerve blocks are not considered in this review. Neither are trials using different types of drugs or techniques with one type of anaesthesia. The primary outcome was mortality. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a nine item scale, and extracted data. The other two reviewers independently checked these results. Wherever appropriate and possible, results were pooled. MAIN RESULTS: Fifteen trials, involving 2162 patients, which compared regional anaesthesia with general anaesthesia, were included. All trials had methodological flaws. Regional anaesthesia was associated with a decreased mortality at one month (49/766 (6.4%) versus 76/812 (9.4%)) of borderline statistical significance (Peto odds ratio 0.66, 95% confidence interval 0.46 to 0.96)). The results for three month mortality were not statistically significant, although the confidence interval does not exclude the possibility of a clinically relevant reduction (86/726 (11.8%) versus 98/765 (12.8%), Peto odds ratio 0.91, 95% confidence interval 0.67 to 1.24). The reduced numbers at one year, coming exclusively from two studies, preclude any useful conclusions for long term mortality (80/354 (22.6%) versus 78/372 (21.0%), Peto odds ratio 1.10, 95% confidence interval 0.77 to 1.57). Regional anaesthesia was associated with a tendency to a longer operation (weighted mean difference 4.8 minutes, 95% confidence interval 1.1 to 8.6 minutes), and a reduced risk of deep venous thrombosis (39/129 (30%) versus 61/37(76%); Peto odds ratio 0. 41, 95% confidence interval 0.23 to 0.72), although this conclusion is insecure due to possible selection bias in the subgroups in whom this outcome was measured. No other statistically significant differences in outcome were identified. There was insufficient evidence to draw any conclusions from a further two included trials, involving a total of 100 patients, which compared other types of anaesthesia. REVIEWER'S CONCLUSIONS: Regional anaesthesia and general anaesthesia appear to produce comparable results for most of the outcomes studied. Regional anaesthesia may reduce short-term mortality but no conclusions can be drawn for longer term mortality. PMID- 10796388 TI - Osteotomy, compression and reaming techniques for internal fixation of extracapsular hip fractures. AB - BACKGROUND: Many different surgical techniques, such as osteotomy, have been used in internal fixation of extracapsular hip fractures. OBJECTIVES: To compare different aspects of surgical technique in internal fixation of extracapsular hip fractures that have been subjected to randomised trials in adults. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group trials register, Medline, CENTRAL and reference lists of relevant articles were searched. Date of the most recent search: March 1999. SELECTION CRITERIA: All randomised and quasi randomised trials investigating operative technique for the treatment of extracapsular hip fractures. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, by use of an eleven item check list, and extracted data. Wherever possible and appropriate, results of outcome measures were pooled. MAIN RESULTS: All eight included trials were of only modest methodological quality. One trial of 65 patients undergoing fixation with a fixed nail-plate compared osteotomy versus anatomical reduction. There was a tendency to a reduced fixation failure rate after osteotomy. Four trials involving 465 patients undergoing fixation with a sliding hip screw (SHS) compared osteotomy versus anatomical reduction. Osteotomy was associated with an increased operative blood loss and length of surgery. There was also a tendency to an increased length of hospital stay and limb shortening for osteotomy. One trial of 200 patients undergoing fixation with a SHS compared results with or without compression across the fracture site. The only significant difference in outcomes was increased varus deformity in those fractures treated with compression. One trial of 19 patients reported reduced temperatures generated by a modified method of reaming the femoral head. Another study used oesophageal ultrasound to demonstrate reduced bone marrow intravascular embolism when a Gamma nail was inserted in 50 patients with, rather than without, a distal pressure venting hole in the femur. REVIEWER'S CONCLUSIONS: There is inadequate evidence to determine if any benefits exist for the routine use of osteotomy in conjunction with a SHS for the internal fixation of an unstable trochanteric femoral fracture. Osteotomy may be relevant if used in conjunction with a fixed nail plate. Based on the evidence of one trial only, there is inadequate evidence to support the application of compression across the fracture site of a trochanteric fracture during SHS fixation. Inadequate information exists for different reaming techniques during SHS or Gamma nail fixation to make definite conclusions. PMID- 10796389 TI - Interventions for isolated diaphyseal fractures of the ulna in adults. AB - BACKGROUND: Isolated fractures of the shaft of the ulna, which are often sustained when the forearm is raised to shield against a blow, are generally treated on an out-patient basis. OBJECTIVES: To assess the effects of various forms of treatment for isolated fractures of the ulnar shaft in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, MEDLINE (1976 to August 1999), EMBASE (1981 to September 1999), the Cochrane Controlled Trials Register (up to Issue 4, 1999), and bibliographies of trial reports. Date of the most recent search: August 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials of conservative and surgical treatment of isolated fractures of the ulnar shaft in adults. Excluded were fractures of the proximal ulna and Monteggia fracture dislocations. DATA COLLECTION AND ANALYSIS: Independent quality assessment and data extraction were performed by all reviewers. Requests for more information were sent to trialists. Given the limited and poor quality evidence available, quantitative analysis was kept to a minimum. MAIN RESULTS: Two small trials of conservative treatment, involving a total of 106 patients were included in this review. Both trials were of poor quality. One randomised trial compared short arm pre-fabricated functional braces with long arm plaster casts. There was no statistically significant difference in the time it took for fracture union. Patient satisfaction and return to work were better in the brace group. The other quasi-randomised trial compared Ace Wrap elastic bandage, short arm plaster cast and long arm plaster cast. The large loss to follow-up in this trial makes any data analysis tentative. However the need for replacement of the Ace wrap by other methods due to pain does indicate the potential for a serious problem with this intervention. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to determine which method of treatment is the most appropriate for isolated fractures of the ulnar shaft in adults. There is a need to establish the incidence of this injury and the outcome and associated costs of the various forms of treatment. Well designed randomised trials of current forms of conservative treatment are recommended. PMID- 10796390 TI - Cognitive behaviour therapy for schizophrenia. AB - BACKGROUND: Although medication is the mainstay of treatment for schizophrenia, always, some sort of informal or formal talking therapy is indicated. In cognitive behavioural therapy (CBT) links are made between the person's feelings and patterns of thinking which underpin their distress. OBJECTIVES: To review the effects of cognitive behaviour therapy (CBT) for those with schizophrenia compared to standard care, specific medication and non-intervention; also to review the effects of CBT for those with schizophrenia who are concurrently receiving standard care compared to no additional intervention to standard care, specific medication, additional drug interventions to standard care and other additional psychosocial interventions to standard care. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1998), CINAHL (1982-1998), The Cochrane Library (Issue 2, 1998), The Cochrane Schizophrenia Group's Register of Trials (August 1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1887 1998), SIGLE (1990-1998), and Sociofile (1980-1998) were undertaken. All references of articles selected were searched for further relevant trials. SELECTION CRITERIA: Randomised trials of cognitive behaviour therapy for people with a diagnosis of schizophrenia, possible schizophrenia or mental illnesses where specific diagnoses have not been employed. Outcomes such as death, metal state, relapse, psychological well-being and acceptability of treatment were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected and assessed for methodological quality. Data were extracted by two reviewers working independently. Dichotomous data were analysed on an intention-to-treat basis and continuous data with 70% completion rate are presented. MAIN RESULTS: Four small trials were identified. All presented data suggested that there was a difference favouring CBT plus standard care over standard care alone in terms of reducing relapse rates (short term OR 0.31 CI 0.1-0.98; medium term OR 0.38 CI 0.17-0.83; long term OR 0.46 CI 0.26-0.83, NNT 6 CI 3-30). These findings were supported within the trials by scale-derived data. CBT, however, did not keep more people in care than a standard approach and there is no data relating to the effect of CBT on compliance with medication. One study also presented data on the effects of CBT when compared to supportive psychotherapy. No effect statistically significantly favoured either group but all were suggestive that the trial may have been underpowered to find an effect in favour of CBT. REVIEWER'S CONCLUSIONS: The results of well conducted and reported ongoing trials are eagerly awaited. Currently, for those with schizophrenia willing to receive CBT, access to this treatment approach is associated with a substantially reduced risk of relapse. However, at present CBT is a fairly scarce commodity, often provided by highly skilled and experienced therapists. Therefore, its application in day to day practice may be restricted by the availability of suitable practitioners. Similarly, the present data provides little indication of how effective CBT procedures might be when they are applied by less experienced practitioners. PMID- 10796391 TI - Nasal versus oral intubation for mechanical ventilation of newborn infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: The purpose of the review was to compare the complications associated with intubation by the nasal route with those associated with intubation by the oral route for mechanical ventilation in newborn infants. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group as outlined in the Cochrane Library was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Register of Controlled Trials, MEDLINE and CINAHL. A call was placed on the list servers, NICU-NET and Neonatal Talk for unpublished trials, conference presentations and current trials. SELECTION CRITERIA: All trials using random or quasi-random allocation of patients to either the nasal or oral route of intubation were included. Study quality and eligibility were assessed independently by each author. DATA COLLECTION AND ANALYSIS: The standard method of the Cochrane Collaboration and the Neonatal Review Group was used to assess the methodological quality of the included studies. The methodological quality of each study was reviewed by the second author blinded to study authors and institutions. Each reviewer extracted data separately before comparison and resolution of differences. The standard method of the Neonatal Review Group was used to measure the effect of the different routes of intubation, using Relative Risk (RR) and 95% Confidence Intervals (CI). MAIN RESULTS: Only two eligible randomized trials were found. Data from these two trials failed to show significant differences between the oral and nasal route of intubation for mechanically ventilated neonates. The rate of failure to intubate using the nasal route was higher in one study. One study found post extubation atelectasis occurred more frequently in nasally intubated infants who weighed less than 1500 grams. The rates of malposition of the tube at the initial intubation, accidental extubation, tube blockage, re-intubation after extubation, septicaemia, clinical infection and local trauma (nasal erosion or palatal groove) were not significantly different for the two groups. REVIEWER'S CONCLUSIONS: Post extubation atelectasis may be more frequent after nasal intubation, particularly in very low birth weight infants. One route of intubation does not seem to be preferable to the other. There is a need for further randomized controlled trials containing larger numbers of infants. PMID- 10796392 TI - Umbilical artery catheters in the newborn: effects of catheter materials. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine whether the material used for construction of an umbilical arterial catheter influences the frequency of ischemic events, aortic thrombosis, mortality or necrotising enterocolitis in newborn infants. SEARCH STRATEGY: Randomized and quasi randomized controlled trials of umbilical catheterization use were obtained from the following sources: 1. Effective Care of the Newborn Infant, edited by JC Sinclair and MB Bracken. 2. Medline Search using Melvyl Medline Plus and the keyword headings "Umbilic#", "Catheter#" and subject heading "Infant, Newborn" 3. Search of personal data files SELECTION CRITERIA: Randomized studies in newborn infants of any birthweight or gestation. Comparison of different catheter materials. Clinically important end points such as ischemic events, aortic thrombosis, or catheter occlusion. DATA COLLECTION AND ANALYSIS: One non-randomized and one randomized study were retrieved. MAIN RESULTS: There were no significant effects of substituting a heparin bonded polyurethane catheter for the standard PVC catheter. The non-randomized study suggested that there may possibly be a benefit of using a catheter constructed from Silastic, with a reduction in aortic thrombosis. REVIEWER'S CONCLUSIONS: There are no demonstrated clinically relevant differences in outcomes between the use of PVC catheters and other materials. Therefore, other considerations such as price and ease of availability may dictate the catheter chosen. An adequately powered randomized comparison of silastic to PVC should be performed. PMID- 10796393 TI - Folic acid and folinic acid for reducing side effects in patients receiving methotrexate for rheumatoid arthritis. AB - OBJECTIVES: To assess the effects of folic acid and folinic acid in reducing the mucosal and gastrointestinal (GI) and haematologic side effects of low-dose of Methotrexate (MTX) in patients with Rheumatoid Arthritis (RA) and to determine whether or not folate supplementation alters MTX efficacy. SEARCH STRATEGY: We searched the Cochrane Controlled Clinical Trial's Register (CCTR), the Cochrane Musculoskeletal Group Specialized Register and Medline up to and including June 1999, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). We also handsearched the following: (i) bibliographic references; (ii) current contents of the last 6 months; (iii) abstracts of the rheumatology meetings; and (iv) all issues of four journals; Journal of Rheumatology, Arthritis & Rheumatism, Clinical and Experimental Rheumatology, and British Journal of Rheumatology. All languages were included. Principal investigators were also contacted in order to look for unpublished literature. SELECTION CRITERIA: We selected all double-blind, randomized, placebo-controlled, clinical trials (RCTs), in which adult RA patients were treated with a low dose of MTX (<20 mg / week) concurrently with folate supplementation. DATA COLLECTION AND ANALYSIS: Two observers extracted the data and assessed the quality of the trials. (BS, Z0) The overall treatment effect across trials was calculated using a fixed effect model. Disease activity was evaluated using standardized mean differences to ensure comparability across outcome measures. Results are presented with 95% Confidence Inervals (95% CI). Subgroup analyses were conducted evaluating different doses and sensitivity analysis looking at the quality of the trials. Publication bias was assessed with an inverted funnel plot technique. Heterogeneity of the trials was measured using a standard chi square test. Costs per month in different countries were compared. MAIN RESULTS: Of the 12 trials retrieved, 7 met the inclusion criteria. The total sample included 307 patients, of which 147 were treated with folate supplementation, 80 patients with folinic acid and 67 patients with folic acid. A 79% reduction in mucosal and GI side effects was observed for folic acid [OR = 0.21 (95% CI 0.10 to 0.44)]. For folinic acid, a clinically but non-statistically significant reduction of 43% was found [OR = 0. 57 (95% CI 0.28 to 1.15)]. No major differences were observed between low and high doses of folic or folinic acid. Haematologic side effects could not be analyzed, since details of each haematologic side effect by patients were not provided. No consistent differences in disease activity parameters were observed when comparing placebo and folic or folinic acid at low or high doses, although patients on high dose folinic acid had an increase in the number of tender joints, but not swollen joints. Large differences in costs across countries were found, but folinic acid was more expensive in all. REVIEWER'S CONCLUSIONS: The results support the protective effect of folate supplementation in reducing MTX side effects related to the oral and GI systems. We could not determine if folic was different from folinic acid. Therefore, for folinic acid to be considered cost-effective it must be found more effective than folic acid at reducing MTX side effects. PMID- 10796394 TI - Calcium and vitamin D for corticosteroid-induced osteoporosis. AB - OBJECTIVES: To assess the effects of calcium and vitamin D compared to calcium alone or placebo in the prevention of bone loss in patients taking systemic corticosteroids. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal trials register, Cochrane Controlled Trials Register, EMBASE and Medline up to 1996. We also conducted a hand search of abstracts from various scientific meetings and reference lists of selected trials. SELECTION CRITERIA: All randomized trials comparing calcium and vitamin D to calcium alone or placebo in patients taking systemic corticosteroids. DATA COLLECTION AND ANALYSIS: Data was abstracted from trials by two investigators. Methodological quality was assessed in a similar manner. Analysis was performed using fixed effects models. MAIN RESULTS: Five trials were included, with 274 patients. The analysis was performed at two years after starting calcium and vitamin D. There was a significant weighted mean difference (WMD) between treatment and control groups in lumbar (WMD 2.6 (95% CI 0.7, 4.5), and radial bone mineral density (WMD 2.5 (95% CI 0.6, 4.4). The other outcome measures (femoral neck bone mass, fracture incidence, biochemical markers of bone resorption) were not significantly different. REVIEWER'S CONCLUSIONS: This meta-analysis demonstrated a clinically and statistically significant prevention of bone loss at the lumbar spine and forearm with vitamin D and calcium in corticosteroid treated patients. Because of low toxicity and cost all patients being started on corticosteroids should receive prophylactic therapy with calcium and vitamin D. PMID- 10796395 TI - Iloprost and cisaprost for Raynaud's phenomenon in progressive systemic sclerosis. AB - OBJECTIVES: To assess the effects and toxicity of the following agents:Prostaglandin analogues together with other agents proposed for the treatment of Raynaud's phenomenom (RP) in scleroderma. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, and Medline up to 1996 using the Cochrane Collaboration search strategy developed by Dickersin et al.(1994). Key words included: raynaud's or vasospasm, scleroderma or progressive systemic sclerosis or connective tissue disease or autoimmune disease. Current Contents were searched up to and including April 7, 1997. All bibliographies of articles retrieved were searched and key experts in the area were contacted for additional and unpublished data. The initial search strategy included all languages. SELECTION CRITERIA: All randomized controlled trials comparing prostaglandin analogues versus placebo were eligible if they reported clinical outcomes within the start of therapy, and if the dropout rate was less than 35%. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by two reviewers (DF, AT). Peto's odds ratios were calculated for all dichotomous outcomes and a weighted mean difference was calculated for all continuous outcomes. A fixed effects or random effects model was used if the data were homogeneous or heterogeneous, respectively. MAIN RESULTS: Seven randomized trials and 332 patients were included. Five of the seven trials were of parallel design. Five trials compared I.V. Iloprost and one trial studied p.o. Iloprost and another p.o. Cisaprost. Some trials were dose finding trials so various doses of Iloprost were used. Due to different efficacies of I.V. Iloprost, oral Iloprost and oral Cisaprost, the overall efficacy of these drugs was somewhat diluted. Intravenous Iloprost appears to be effective in the treatment of secondary Raynaud's phenomenon. REVIEWER'S CONCLUSIONS: Intravenous Iloprost is effective in the treatment of Raynaud's phenomenon secondary to scleroderma at decreasing the frequency and severity of attacks and preventing or healing digital ulcers. The effect seems to be prolonged after the intravenous infusion is given. Oral Iloprost may have less efficacy than intravenous Iloprost. However, Cisaprost has minimal or no efficacy when given orally for the treatment of Raynaud's phenomenon secondary to scleroderma. PMID- 10796396 TI - Ketanserin for Raynaud's phenomenon in progressive systemic sclerosis. AB - OBJECTIVES: To assess the effects and toxicity of the following agent: ketanserin versus placebo proposed for the treatment of Raynaud's phenomenon (RP) in scleroderma. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, and Medline up to 1996 using the Cochrane Collaboration search strategy developed by Dickersin et al.(1994). Key words included: Raynaud's or vasospasm, scleroderma or progressive systematic sclerosis or connective tissue disease or autoimmune disease. Current Contents were searched up to and including April 7, 1997. All bibliographies of articles retrieved were searched and key experts in the area were contacted for additional and unpublished data. The initial search strategy included all languages. SELECTION CRITERIA: All randomized controlled trials comparing ketanserin versus placebo were eligible if they reported clinical outcomes of interest. Trials with dropout rates greater than 35% were excluded. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by two reviewers (DF, AT). Peto's odds ratios (OR) were calculated for all dichotomous outcomes, and a weighted mean difference (WMD) was carried out on all continuous outcomes. A fixed effects or random effects model were used if the data was homogeneous or heterogeneous, respectively. MAIN RESULTS: Three trials and 66 patients were included. The proportion improved was significantly better in the group on ketanserin with an odds ratio (OR) of 4.80 (95% CI 1.33, 17.37). However, when comparing ketanserin to placebo, the decrease in severity of RP attacks favoured placebo but this was not statistically significant. Side effects were significantly more common in the group using active treatment with an OR of 5.96 (95% CI 1.61, 22.06). Frequency of attacks did not change, but the duration of attacks decreased significantly in the ketanserin group. REVIEWER'S CONCLUSIONS: Ketanserin may have some efficacy in the treatment of Raynaud's phenomenon secondary to scleroderma. Overall, ketanserin is not significantly different from placebo for the treatment of Raynaud's phenomenon except for some decrease in the duration of attacks and more subjects improved on ketanserin compared to placebo. However, there were more side effects. It can be concluded that ketanserin treatment in Raynaud's phenomenon secondary to scleroderma is not clinically beneficial. PMID- 10796397 TI - Cyclofenil for Raynaud's phenomenon in progressive systemic sclerosis. AB - OBJECTIVES: To assess the effects and toxicity of Cyclofenil versus placebo for the treatment of Raynaud's phenomenon (RP) in scleroderma. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, and Medline up to 1996 using the Cochrane Collaboration search strategy developed by Dickersin et al.(1994). Key words included: Raynaud's or Vasospasm, Scleroderma or Progressive Systematic Sclerosis or Connective Tissue Disease or Autoimmune Disease. Current Contents were searched up to and including April 7, 1997. All bibliographies of articles retrieved were searched and key experts in the area were contacted for additional and unpublished data. The initial search strategy included all languages. SELECTION CRITERIA: All randomized trials comparing cyclofenil versus placebo were eligible if they reported any clinical outcomes within the trial. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by two reviewers (DF, AT). Peto's odds ratio (OR) was calculated for all dichotomous outcomes, and a weighted mean difference was calculated for all continuous outcomes. A fixed effects or random effects model was used if the data were homogeneous or heterogeneous respectively. MAIN RESULTS: One trial with 38 patients was included. There was a trend for Cyclofenil to demonstrate more improvement and more dropouts compared to placebo, but there were no statistically significant differences. REVIEWER'S CONCLUSIONS: Cyclofenil is not effective in the treatment of Raynaud's phenomenon secondary to scleroderma. PMID- 10796398 TI - Prazosin for Raynaud's phenomenon in progressive systemic sclerosis. AB - OBJECTIVES: To determine the effects and toxicity of prazosin versus placebo proposed for the treatment of Raynaud's phenomenon (RP) in scleroderma. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, and Medline up to December 1996 using the Cochrane Collaboration search strategy developed by Dickersin et al.(1994). Key words included: Raynaud's or vasospasm, scleroderma or progressive systemic sclerosis or connective tissue disease or autoimmune disease. Current Contents were searched up to and including April 7, 1997. All bibliographies of articles retrieved were searched and key experts in the area were contacted for additional and unpublished data. The initial search strategy included all languages. SELECTION CRITERIA: Randomized controlled trials comparing prazosin versus placebo were eligible if they reported clinical outcomes from the start of therapy. Trials with a greater than 35% dropout were excluded. Trials were included if patients with diffuse or limited scleroderma were the subjects. If patients with other connective tissue diseases or primary Raynaud's were included, the trial was used if the data on the scleroderma patients could be extracted from the paper. DATA COLLECTION AND ANALYSIS: All data were abstracted by two independent and trained reviewers (DF, AT), and verified by a third reviewer (JP). Each trial was assessed independently by the same two reviewers for its quality using a validated quality assessment tool (Jadad 1996). Peto's odds ratios were calculated for all dichotomous outcomes and a weighted mean difference was carried out on all continuous outcomes. Fixed effects and random effects model were used if the data was homogeneous or heterogeneous, respectively. MAIN RESULTS: Two trials with a total of 40 patients were included. Prazosin has been found in two randomized controlled cross-over trials to be more effective than placebo in the treatment of Raynaud's secondary to scleroderma. However, the positive response is modest and side effects are not rare in those taking prazosin. REVIEWER'S CONCLUSIONS: Prazosin is modestly effective in the treatment of Raynaud's phenomenon secondary to scleroderma. PMID- 10796399 TI - Methotrexate for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term efficacy and toxicity of methotrexate (MTX) for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, and Medline, up to July 1997, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. SELECTION CRITERIA: Randomized controlled trials and controlled clinical trials comparing MTX against placebo in patients with RA. DATA COLLECTION AND ANALYSIS: Two reviewers determined the studies to be included based on inclusion and exclusion criteria (GW, MSA). Data were independently abstracted by two reviewers (EB, MSA), and checked by a third reviewer (BS) using a pre-developed form for the rheumatoid arthritis sub-group of the Cochrane Musculoskeletal Group. The same two reviewers, using a validated scale (Jadad 1996) assessed the methodological quality of the trials independently. Rheumatoid arthritis outcome measures were extracted from the publications. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts, pain, and global and functional assessments. Weighted mean differences (WMDs) were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios (OR) for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout and random effects for outcomes showing heterogeneity. MAIN RESULTS: Five trials and 300 patients were included. A statistically significant benefit was observed for MTX when compared to placebo. Statistically significant differences were observed for all measures except ESR. The standardized weighted difference (effect size) between MTX and placebo for the various outcome measures varied between -0.43 and -1.5. No differences were observed in the total number of withdrawals and dropouts (OR = 0.95), although patients on MTX were three times more likely to discontinue treatment because of adverse reactions (OR=3.47) and four times less likely to withdraw due to lack of response (OR=0.22). REVIEWER'S CONCLUSIONS: Twenty-two percent of people on MTX withdrew due to adverse effects compared to seven percent of the placebo group. MTX has a substantial clinically and statistically significant benefit in the short term treatment of patients with RA. PMID- 10796400 TI - Sulfasalazine for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term efficacy and toxicity of sulfasalazine for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, and Medline, up to July 1997, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing sulfasalazine against placebo in patients with RA. DATA COLLECTION AND ANALYSIS: Two reviewers determined the studies to be included based on inclusion and exclusion criteria (GW, MSA). Data were independently abstracted by two reviewers (EB, MSA), and checked by a third reviewer (BS) using a pre-developed form for the rheumatoid arthritis sub-group of the Cochrane Musculoskeletal Group. The same two reviewers, using a validated scale (Jadad 1996) assessed the methodological quality of the RCTs and CCTs independently. Rheumatoid arthritis outcome measures were extracted from the publications. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts, pain, and global and functional assessments. Weighted mean differences (WMDs) were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios (OR) for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout and random effects for outcomes showing heterogeneity. MAIN RESULTS: Six trials, including 468 patients were included. A statistically significant benefit was observed for sulfasalazine when compared to placebo for tender and swollen joint scores, pain and ESR. The standardized weighted mean difference between treatment and placebo was -0.49 for tender and swollen joint scores, and 0.42 for pain. The difference for ESR was -17.6mm. Withdrawals from adverse reactions were significantly higher in the sulfasalazine group (OR=3.0). Patients receiving placebo were four times more likely to discontinue treatment because of lack of efficacy than patients receiving sulfasalazine. REVIEWER'S CONCLUSIONS: Sulfasalazine appears to have a clinically and statistically significant benefit on the disease activity of patients with RA. Its effects on overall health status and radiological progression are not clear at this time, but would appear to be modest. PMID- 10796401 TI - Antimalarials for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term efficacy and toxicity of antimalarials for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, Medline and Embase up to and including July 1997. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing antimalarials against placebo in patients with RA DATA COLLECTION AND ANALYSIS: Data abstraction was carried out independently by two reviewers. The same two reviewers using Jadad's scale (Jadad 1995) assessed the methodological quality of the RCTs and CCTs. Rheumatoid arthritis outcome measures were extracted from the publications for the 6-month endpoint. The pooled analysis was performed using standardized mean differences for joint counts, pain and global assessments. Weighted mean differences were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout. MAIN RESULTS: We found four trials, with 300 patients randomized to hydrochloroquine and 292 to placebo. Only trials evaluating hydroxychloroquine could be pooled in the analysis. A statistically significant benefit was observed when hydroxychloroquine was compared to placebo. The standardized mean differences for the various outcome measures ranged from -0.33 to -0. 52, and were statistically significant. Statistically significant differences were also observed for ESR. Overall withdrawals and withdrawals due to lack of efficacy were significantly more frequent in the placebo group. No differences were observed in withdrawals due to toxicity. REVIEWER'S CONCLUSIONS: Hydroxychloroquine appears to be efficacious for the treatment of RA. Its overall effect appears to be moderate, but its low toxicity profile should be considered when treating patients with RA. PMID- 10796402 TI - Physical medicine modalities for mechanical neck disorders. AB - OBJECTIVES: Multiple physical medicine modalities are commonly included as part of therapeutic interventions for mechanical neck disorders (neck pain). The objective of this review was to assess the effects of physical medicine modalities for pain in adults with mechanical neck disorders. SEARCH STRATEGY: We searched Medline, Embase, Chirolars, Index to Chiropractic Literature, Cinahl, Science Citation Index, Conference Proceedings Index, National Technical Information Services and reference lists of the retrieved articles from 1985 to December 1993 and we contacted content experts. SELECTION CRITERIA: Randomised trials and controlled trials of physical medicine modalities in adults with mechanical neck disorder. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality and two reviewers independently extracted data. Investigators were contacted to obtain information or data that could not be found in the published reports. MAIN RESULTS: Thirteen trials were included. The overall quality of the included trials was generally good. Two trials using electromagnetic therapy produced a significant reduction in pain (p <0.01) with three to four weeks of daily (eight hours per day) therapy sessions; and three using laser therapy did not differ significantly from a placebo (p=0.20) for six to 10 sessions of treatment. Not enough scientific testing exists to clearly determine the effectiveness of other therapies. This includes treatments such as exercise, traction, acupuncture, heat / cold applications, electrotherapies, cervical orthoses and chronic pain / cognitive behavioural rehabilitation strategies. REVIEWER'S CONCLUSIONS: There is little information available from trials to support the use of physical medicine modalities for mechanical neck pain. There is some support for the use of electromagnetic therapy and against the use of laser therapy with respect to pain reduction. PMID- 10796403 TI - Patient education for mechanical neck disorders. AB - BACKGROUND: To track down the best estimate of efficacy of the various conservative management strategies for mechanical neck disorders, a four-part systematic review was prepared. Part four investigates the efficacy of patient education strategies as the therapeutic intervention. OBJECTIVES: This review of patient education is one of four reviews of conservative management of mechanical neck disorders. The other reviews address manual, physical and drug therapies. The objective of this review was to assess the effects of patient education for pain in adults with mechanical neck disorders. SEARCH STRATEGY: We searched Medline, Embase, Chirolars, Index to Chiropractic Literature, Cinahl, Science Citation Index, Conference Proceedings Index, National Technical Information Services from 1985 to December 1993, reference lists of the retrieved articles and we contacted experts in the field. SELECTION CRITERIA: Randomised trials or controlled clinical trials of patient educational strategies for adults with mechanical neck disorders. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality and two reviewers independently extracted data. Investigators were contacted to obtain data that could not be found in the published reports. MAIN RESULTS: Three trials were included. Their methodological quality varied from weak to strong. One trial did not find a significant reduction in pain using group instructional strategies (neck school) and exercise with or without psychological counselling compared to no treatment (standardised mean difference 0.07, 95% confidence interval -0.51 to 0.66, and -0.37, 95% confidence interval -0.95 to 0.22, respectively). Another trial did not find a significant reduction in pain using individualised patient education (advice), anti-inflammatories and analgesics compared with placebo (standardised mean difference 0.24, 95% confidence interval -0.58 to 1.07). The third trial found that advice which included demonstrated mobilization exercises, verbal and written instruction on posture correction, the use of a collar, heat sources, muscle relaxation and analgesics gave significant pain relief compared with general advice about mobilisation after a period of rest and use of analgesics at 4 weeks of treatment (standardised mean difference -0.62, 95% confidence interval -1.05 to -0.19) but at 6 weeks of treatment there was no longer any difference (s.m.d. -0.37, 95% confidence interval -0.8 to 0.05). The first two trials lacked statistical power and the third was methodologically weak. REVIEWER'S CONCLUSIONS: Patient education utilising individualised or group instructional strategies has not been shown to be beneficial in reducing pain for mechanical neck disorders. PMID- 10796404 TI - Interventions for treating hallux valgus (abductovalgus) and bunions. AB - BACKGROUND: Hallux valgus is classified as an abnormal deviation of the great toe (hallux) towards the midline of the foot. OBJECTIVES: To identify and evaluate the evidence from randomised trials of interventions used to correct hallux valgus. SEARCH STRATEGY: Medline (1966-October 1998), Embase (1980-1998), Cinahl (1982-1998), Amed (1993-1998), the Cochrane Controlled Trials Register, the Cochrane Musculoskeletal Injuries Trials Register and bibliographies of identified trials were searched. Hand searching of podiatry journals was undertaken. Date of the most recent search: October 1998. SELECTION CRITERIA: Randomised or quasi-randomised trials of both conservative and surgical treatments of hallux valgus. Excluded were studies comparing areas of surgery not specific to the control of the deformity such as use of anaesthetics or tourniquet placement. DATA COLLECTION AND ANALYSIS: Methodological quality of trials which met the inclusion criteria was independently assessed by two reviewers. Data extraction was undertaken by one reviewer and confirmed by another. The trials were grouped according to the interventions being compared, but the dissimilarity in the comparisons prevented pooling of results. MAIN RESULTS: The methodological quality of the 12 included trials was poor and trial sizes were small. Two trials involving 150 patients evaluated conservative treatments. There was no difference in outcomes between treatment and no treatment. Two trials involving 133 patients compared Keller's arthroplasty with other surgical techniques. In general, there was no advantage in using Keller's over the other techniques, particularly in terms of range of motion and intermetatarsal angle. Three trials involving 205 patients compared chevron (and chevron-type) osteotomy with other techniques. The chevron osteotomy offered no advantages in these trials. For some outcomes, other techniques gave better results. Three trials involving 157 patients compared outcomes between original operations and surgeon's adaptations. There was no advantage found for any of the adaptations. Two trials involving 95 patients evaluated methods of post-operative rehabilitation. The use of continuous passive motion appeared to give an improved range of motion and earlier recovery following surgery; early weightbearing was not found to be detrimental to final outcome. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to determine which methods of either conservative, operative or post-operative treatment are the most appropriate for the hallux valgus. It is notable that the numbers of patients remaining dissatisfied at follow-up were consistently high (25 to 33%), even when the hallux valgus angle and pain had improved. Assessment of future research should focus on evaluating basic intervention types in eligible patients with similar degrees of deformity. Future research should include patient-focused outcomes, standardised assessment criteria and longer surveillance periods. PMID- 10796405 TI - Sucrose for analgesia in newborn infants undergoing painful procedures. AB - BACKGROUND: Management of pain for neonates is less than optimal. The administration of sucrose with and without non-nutritive sucking (pacifiers) has been the most frequently studied non-pharmacological intervention for relief of procedural pain in neonates. OBJECTIVES: To determine the efficacy, effect of dose, and safety of sucrose for relieving procedural pain as assessed by physiologic and/or behavioural indicators. SEARCH STRATEGY: Standard methods as per the Neonatal Collaborative Review Group. A MEDLINE search was carried out for relevant randomized controlled trials (RCTs) published from January 1966 - November 1, 1997; EMBASE from 1993-1997; Reference Update search on November 11, 1997 and search of the Cochrane Library Issue 4 on November 11, 1997. Key words and (MeSH) terms included, infant/newborn, pain, analgesia and sucrose. Personal files, bibliographies, the most recent relevant neonatal and pain journals and conference proceedings were searched manually. Unpublished studies were not included. Language restrictions were not imposed. SELECTION CRITERIA: RCTs in which term and/or preterm neonates undergoing heel lance, venepuncture or intramuscular injection (immunization) received sucrose or water/placebo or no intervention. DATA COLLECTION AND ANALYSIS: Trial quality was assessed according to the methods of the Neonatal Collaborative Review Group. Quality measures included; blinding of randomization, blinding of intervention, completeness of follow up and blinding of outcome measurement. Data were abstracted and independently checked for accuracy by the two investigators. The inconsistency in outcome measures and differences in the statistical reporting of results made meta-analysis impossible. We were not able to identify two studies in which the same physiologic and/or behavioral outcomes following a noxious stimulus (heel lance, venepuncture, intramuscular injection) were measured and reported in an identical fashion using means and standard deviations (or standard errors). The results are therefore reported for each accepted study separately. MAIN RESULTS: Fifteen studies were identified for possible inclusion in this systematic review. Five studies were excluded; three RCTs were excluded as in these trials the number of infants randomized to treatment vs. placebo groups were not reported; one study was not an RCT, and in one RCT the neonates did not undergo a painful procedure. Ten RCTs were included in this review. Sucrose in a wide range of dosages was generally found to decrease univariate physiologic (heart rate) and behavioural (the mean percent time crying, total cry duration, duration of first cry, and facial action) pain indicators and multivariate [Premature Infant Pain Profile (PIPP)] pain outcomes in neonates undergoing heelstick or venepuncture. An optimal dose of sucrose to reduce pain associated with procedures in preterm and term neonates could not be identified through this systematic review. REVIEWER'S CONCLUSIONS: Sucrose reduces physiologic and/or behavioral indicators of stress/pain in neonates following procedural pain stimuli (heel lance, venepuncture, immunization). There was inconsistency in the dose of sucrose that was effective and an optimal dose to be used in preterm and/or term infants could not be identified. Considerations for future research are to describe the painful procedure and intervention in detail, to use appropriate sample size to show a statistically significant reduction in pain, to use a multidimensional conceptualization of pain, to select outcome measures that are reliable and valid pain indicators and to account for the variation in the infant's response and context in which the pain is experienced. The use of repeated administrations of sucrose in neonates needs to be investigated. Use of sucrose in neonates that are very low birth weight, unstable and/or ventilated also needs to be addressed. (ABSTRACT TRUNCATED) PMID- 10796406 TI - Thyroid hormone for preventing of neurodevelopmental impairment in preterm infants. AB - BACKGROUND: Observational studies have shown an association between transiently low thyroid hormone levels in preterm infants in the first weeks of life (transient hypothyroxemia) and an abnormal neurodevelopmental outcome. Thyroid hormone therapy might prevent this morbidity. OBJECTIVES: To assess whether thyroid hormone therapy in preterm infants without congenital hypothyroidism results in clinically important changes in neonatal and long term outcomes in terms of both benefits and harms. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences, symposia proceedings, expert informants and journal handsearching in the English language. SELECTION CRITERIA: All trials using random or quasi-random patient allocation, in which thyroid hormone therapy (either treatment or prophylaxis) was compared to a control in premature infants. DATA COLLECTION AND ANALYSIS: Primary clinical outcomes included measures of neurodevelopmental outcome and mortality. Assessment of trial quality, data extraction and synthesis of data, using relative risk (RR) and weighted mean difference (WMD), were performed using standard methods of the Cochrane Collaboration and its Neonatal Review Group. MAIN RESULTS: Eight studies were identified that compared thyroid hormone treatment to control. Four randomized or quasi-randomized studies met inclusion criteria (Chowdhry 1984, Amato 1989, van Wassenaer 1997 and Vanhole 1997). All studies enrolled preterm infants < 32 weeks gestation, but used different timing, dose and duration of treatment with thyroid hormones. Three studies used thyroxine, whereas Amato 1989 used triiodothyronine. Only two studies with neurodevelopmental follow-up were of good methodology (van Wassenaer 1997 and Vanhole 1997). All studies were of small size with the largest, van Wassenaer 1997, enrolling 200 infants. A lack of comparability of data (neurodevelopmental test or timing of follow-up) prevented meta-analytic pooling of the studies for neurodevelopmental outcomes. There was no significant difference in mortality to discharge (typical relative risk 0.74, 95% CI 0.44, 1.26) in infants who received thyroid hormone treatment compared to controls. In individual studies, no significant differences were found in neurodevelopmental outcomes including risk of abnormal neurological outcome, and Bayley Mental or Psychomotor Development Indices. No data were available for the incidences of cerebral palsy or sensorineural impairment. Fraction of inspired oxygen was lower in infants receiving triiodothyronine in one small quasi-randomized study (Amato 1989), but not in infants receiving thyroxine in a randomized study (Vanhole 1997). No other differences were found to suggest a reduced severity of respiratory distress syndrome in infants receiving early thyroid hormone therapy. REVIEWER'S CONCLUSIONS: This review does not support the use of thyroid hormones in preterm infants to reduce neonatal mortality, improve neurodevelopmental outcome or to reduce the severity of respiratory distress syndrome. The a posteriori subgroup analyses of data from one study (van Wassenaer 1997) which showed benefits in infants 24-25 weeks gestation should be treated with caution. The small number of infants included in trials incorporated in this review limits the power of the meta-analysis to detect clinically important differences in neonatal outcomes. Future trials should be of sufficient size to detect clinically important differences in neurodevelopmental outcomes. They should consider enrolling those infants most likely to benefit from thyroid hormone treatment such as infants born at less than 27 weeks gestation and use thyroid hormones as treatment instead of prophylaxis. PMID- 10796407 TI - Non-nutritive sucking for promoting physiologic stability and nutrition in preterm infants. AB - BACKGROUND: Non-nutritive sucking is used during gavage feeding and in the transition from gavage to breast/bottle feeding in preterm infants. The rationale for this intervention is that non-nutritive sucking facilitates the development of sucking behaviour and improves digestion of enteral feedings. Non-nutritive sucking has been considered to be a benign intervention, although it has the potential to have a negative effect on breastfeeding or on the incidence of later oral aversion. OBJECTIVES: To determine whether non-nutritive sucking (NNS) in preterm infants influences: a) weight gain, b) energy intake, c) heart rate, d) oxygen saturation, e) length of hospital stay, f) intestinal transit time, g) age at full oral feeds, or h) any other clinically relevant outcomes. SEARCH STRATEGY: MEDLINE and CINAHL databases back to 1976 and the Cochrane Controlled Trials register were searched. Reference lists/bibliographies of relevant articles and reviews were also searched. A comprehensive list of relevant articles was sent to two major authors in this area. They were asked if they knew of any other published or unpublished studies relevant to the area that had not been included in the original list. SELECTION CRITERIA: All trials utilizing experimental or quasi-experimental designs in which non-nutritive sucking in preterm infants was compared to no provision of non-nutritive sucking. Measured clinically relevant outcomes. Reports were in English or a language for which a translator was available. Computerized searches were conducted by both reviewers. All potentially relevant titles and abstracts identified by either reviewer were extracted. All retrieved articles were assessed for relevance independently by each reviewer, based on a pre-determined set of criteria. The reference lists/bibliographies of each article were reviewed independently for additional relevant titles and were also retrieved and assessed for relevance. Articles that met all relevance criteria were then assessed for methodologic quality based on a predetermined set of criteria. Those articles judged to have the appropriate quality by both reviewers were included in the analysis. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the two authors. No subgroup analyses were performed because of the small number of studies related to the relevant outcomes. MAIN RESULTS: This review consisted of 19 studies, 13 of which were randomized controlled trials. NNS was found to decrease significantly the length of hospital stay in preterm infants. The review did not reveal a consistent benefit of NNS with respect to other major clinical variables (weight gain, energy intake, heart rate, oxygen saturation, intestinal transit time, and age at full oral feeds). The review identified other positive clinical outcomes of NNS: transition from tube to bottle feeds and behavior. No negative outcomes were reported in any of the studies. REVIEWER'S CONCLUSIONS: This review found a significant decrease in length of stay in preterm infants receiving a NNS intervention. The review did not reveal a consistent benefit of NNS with respect to other major clinical variables (weight gain, energy intake, heart rate, oxygen saturation, intestinal transit time, and age at full oral feeds). The review identified other positive clinical outcomes of NNS: transition from tube to bottle feeds and behavior. No negative outcomes were reported in any of the studies. There were also a number of limitations of the presently available evidence related to the design of the studies, outcome variability, and lack of long-term data. Based on the available evidence, NNS in preterm infants would appear to have some clinical benefit. It does not appear to have any short-term negative effects. In view of the fact that there are no long-term data, further investigations are recommended. In order to facilitate meta-analysis of these data, future research in this area should involve outcome measures PMID- 10796408 TI - Servo-control for maintaining abdominal skin temperature at 36C in low birth weight infants. AB - BACKGROUND: Randomized trials have shown that the neonatal mortality rate of low birth-weight babies can be reduced by keeping them warm. For low birth-weight babies nursed in incubators, warm conditions may be achieved either by heating the air to a desired temperature, or by servo-controlling the baby's body temperature at a desired set-point. OBJECTIVES: In low birth weight infants, to determine the effect on death and other important clinical outcomes of targeting body temperature rather than air temperature as the end-point of control of incubator heating. SEARCH STRATEGY: Standard search strategy of the Cochrane Neonatal Collaborative Review Group. SELECTION CRITERIA: Randomized or quasi randomized trials which test the effects of having the heat output of the incubator servo-controlled from body temperature compared with setting a constant incubator air temperature. DATA COLLECTION AND ANALYSIS: Trial methodologic quality was systematically assessed. Outcome measures included death, timing of death, cause of death, and other clinical outcomes. Categorical outcomes were analyzed using relative risk and risk difference. Meta-analysis assumed a fixed effect model. MAIN RESULTS: Compared to setting a constant incubator air temperature of 31.8C, servo-control of abdominal skin temperature at 36C reduces the neonatal death rate among low birth weight infants: relative risk 0.72 (95% CI 0.54, 0.97); risk difference -12.7% (95% CI -1.6, -23.9). This effect is even greater among VLBW infants. REVIEWER'S CONCLUSIONS: During at least the first week after birth, low birth weight babies should be provided with a carefully regulated thermal environment that is near the thermoneutral point. For LBW babies in incubators, this can be achieved by adjusting incubator temperature to maintain an anterior abdominal skin temperature of at least 36C, using either servo-control or frequent manual adjustment of incubator air temperature. PMID- 10796409 TI - Restricted versus liberal oxygen exposure for preventing morbidity and mortality in preterm or low birth weight infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: In preterm or low birth weight infants, does targeting ambient oxygen concentration to achieve a lower versus higher blood oxygen range, or administering restricted versus liberal supplemental oxygen, influence mortality, retinopathy of prematurity, lung function, growth or development? SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. An additional literature search was conducted of the MEDLINE, EMBASE, and CINAHL databases in order to locate any trials in addition to those provided by the Cochrane Controlled Trials Register (CENTRAL/CCTR). SELECTION CRITERIA: All trials in preterm or low birth weight infants utilising random or quasi-random patient allocation, in which ambient oxygen concentrations were targeted to achieve a lower versus higher blood oxygen range, or restricted versus liberal oxygen was administered, were eligible for inclusion. DATA COLLECTION AND ANALYSIS: The methodological quality of the eligible trials was assessed independently by each author for the degree selection, performance, attrition and detection bias. Data were extracted and reviewed independently by the each author. Data analysis was conducted according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: The restriction of oxygen significantly reduced the incidence and severity of retinopathy of prematurity without unduly increasing death rates in the meta-analysis of the five trials included in this review. The one trial that specifically addressed the question of lower versus higher PaO2 found no effect on death, but did not report (in sufficient detail to warrant inclusion) the effect of this intervention on eye or other outcomes. The effects of either of these oxygen administration policies on other clinically meaningful outcomes including chronic lung disease and long term growth, neurodevelopment, lung or visual function were not reported in any of the available trials. REVIEWER'S CONCLUSIONS: The results of this meta-analysis confirm the commonly held view of today's clinicians that a policy of unrestricted, unmonitored oxygen therapy has potential harms, without clear benefits. However, the question of what is the optimal target range for maintaining blood oxygen levels in preterm/LBW infants was not answered by the data available for inclusion in this review. PMID- 10796410 TI - Prophylactic synthetic surfactant for preventing morbidity and mortality in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effect of prophylactic administration of synthetic surfactant in preterm newborns at risk for developing respiratory distress syndrome (RDS). SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactant; limits: age groups, newborn infants; publication type, clinical trial), previous reviews including cross-references, abstracts, conference and symposia proceedings, expert informants, and journal hand searching in the English language. SELECTION CRITERIA: Randomized, controlled trials which compared the effect of prophylactic synthetic surfactant administered to high risk preterm newborns at or shortly after birth in order to prevent respiratory distress syndrome and other complications of prematurity. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including the incidence of pneumothorax, pulmonary interstitial emphysema, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage (any grade and severe intraventricular hemorrhage), bronchopulmonary dysplasia, mortality, bronchopulmonary dysplasia or death, retinopathy of prematurity (any retinopathy, and retinopathy stages 3-4) mortality to one year of age, and cerebral palsy was excerpted from the report of the clinical trials by the reviewer. Data were analyzed according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Studies of prophylactic administration of synthetic surfactant note a variable improvement in the respiratory status and a decrease in respiratory distress syndrome in infants who receive prophylactic synthetic surfactant. The meta-analysis supports a decrease in the risk of pneumothorax (typical relative risk 0.67, 95% CI 0.50, 0.90; typical risk difference -0.05, 95% CI -0.09, -0. 02), a decrease in the risk of pulmonary interstitial emphysema (typical relative risk 0.68, 95% CI 0.50, 0.93; typical risk difference -0.06, 95% CI -0.11, -0.01), and a decrease in risk of neonatal mortality (typical relative risk 0.70, 95% CI 0.58, 0.85; typical risk difference -0.07, 95% CI -0.11, -0.03). No differences were seen in the risk of intraventricular hemorrhage, necrotizing enterocolitis, bronchopulmonary dysplasia, retinopathy of prematurity and cerebral palsy. The meta-analysis supports an increase in the risk of patent ductus arteriosus associated with prophylactic synthetic surfactant administration (typical relative risk 1.11, 95% CI 1.00, 1.22; typical risk difference 0.05, 95% CI 0. 00,0.10), and an increase in the risk of pulmonary hemorrhage (typical relative risk 3.28, 95% CI 1.50, 7.16; typical risk difference 0.03, 95% CI 0.01, 0.05). REVIEWER'S CONCLUSIONS: Prophylactic intratracheal administration of synthetic surfactant to infants judged to be at risk of developing respiratory distress syndrome has been demonstrated to improve clinical outcome. Infants who receive prophylactic synthetic surfactant have a decreased risk of pneumothorax, a decreased risk of pulmonary interstitial emphysema, and a decreased risk of neonatal mortality. Infants who receive prophylactic synthetic surfactant have an increased risk of developing patent ductus arteriosus and pulmonary hemorrhage. PMID- 10796411 TI - Carotid endarterectomy for symptomatic carotid stenosis. AB - BACKGROUND: Severe narrowing (or stenosis) of the carotid artery is an important cause of stroke. Surgical removal of the atheromatous material from the inside of the carotid artery (endarterectomy) may reduce the risk of stroke, but carries a risk of operative complications. OBJECTIVES: This review seeks to summarize the evidence from randomized trials on the balance of risks and benefits of carotid endarterectomy in adults with symptomatic carotid stenosis. SEARCH STRATEGY: We searched the Cochrane Stroke Group's Specialized Register of trials (date last searched: March 1999), supplemented by electronic searches of several databases. SELECTION CRITERIA: Randomized controlled trials comparing 'best medical treatment plus carotid endarterectomy' with 'best medical therapy' in patients with carotid stenosis and a recent transient ischaemic attack or nondisabling ischaemic stroke in the territory of that artery. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected the studies and extracted the data. An intention to treat analysis was performed. MAIN RESULTS: Data on death or disabling stroke were available from two trials, which included 5950 patients: the North American Symptomatic Carotid Endarterectomy Trial (NASCET), and the European Carotid Surgery Trial (ECST). The two trials used different methods to measure stenosis, but a simple formula can be used to convert between the two methods. For patients with severe stenosis (ECST > 80% = NASCET > 70%), surgery reduced the relative risk of disabling stroke or death by 48% (95% confidence interval [CI] 27 - 73%). The number of patients needed to be operated on (number needed to treat [NNT]) to prevent one disabling stroke or death over 2 to 6 years follow-up was 15 (95% CI 10 - 31). For patients with less severe stenosis (ECST 70 - 79% = NASCET 50 - 69%), surgery reduced the relative risk of disabling stroke or death by 27% (95% CI 15 - 44%). The number of patients needed to be operated on to prevent one disabling stroke or death was 21 (95% CI 11 - 125). Patients with lesser degrees of stenosis were harmed by surgery. Surgery increased the risk of disabling stroke or death by 20% (95% CI 0 - 44%). The number of patients needed to be operated on to cause one disabling stroke or death was 45 (95% CI 22 - infinity). REVIEWER'S CONCLUSIONS: Carotid endarterectomy reduced the risk of disabling stroke or death for patients with stenosis exceeding ECST-measured 70% or NASCET-measured 50%. This result is generalizable only to surgically-fit patients operated on by surgeons with low complication rates (less than 6%). PMID- 10796412 TI - Cyclosporine for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term (up to one year) effects of cyclosporine for rheumatoid arthritis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, and Medline, up to 1997, using the search strategy developed by the Cochrane Collaboration (Dickersin 1994). The search was complemented with bibliography searching of the reference list of the trials retrieved from the electronic search. Key experts in the area were contacted for further published and unpublished articles. SELECTION CRITERIA: All randomized clinical trials (RCTs) and controlled clinical trials (CCTs) comparing cyclosporine against placebo in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: Two reviewers determined the trials to be included based on inclusion and exclusion criteria (GW, MSA). Data were independently abstracted by two reviewers (DH, GW),and checked by a third reviewer (BS) using a pre developed form for the rheumatoid arthritis sub-group of the Cochrane Musculoskeletal Group. Methodological quality of the RCTs and CCTs was assessed by two reviewers (BS, DH). Rheumatoid arthritis outcome measures were extracted from the publications for change from baseline endpoints. Sufficient data were obtained to include in the pooled analysis the number of swollen joints, physician global assessment, patient global assessment and erythrocyte sedimentation rate (ESR). MAIN RESULTS: Three trials and 318 patients were included. A statistically significant decrease in the number of tender and swollen joints was observed for cyclosporine when compared to placebo. The standardized mean difference (SMD) for the change in the number of swollen joints was -0.969. Significant improvements in pain and the functional index were also found for cyclosporine. More side effects occurred in the cyclosporine group compared to placebo. REVIEWER'S CONCLUSIONS: Cyclosporine has an important clinical benefit int the short-term (up to one year) treatment of patients with progressive rheumatoid arthritis. PMID- 10796413 TI - Pharmacotherapy for Behcet's syndrome. AB - OBJECTIVES: To determine the effects of available pharmacological interventions in treating the different clinical features of Behcet's syndrome. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, and Medline up to January 1998. The computer search was complemented by a hand search of all bibliographic references from the reference lists of included trials. Principal investigators were contacted to seek unpublished literature. All languages were included. SELECTION CRITERIA: Studies were eligible if they fulfilled all of the four following criteria: 1. Randomized controlled trials, single or double-blind; 2. Participants were patients with Behcet's Syndrome as defined by the International Study Group, 1990 (Int Study Group, 1990); 3. Interventions included any pharmacological therapy compared to placebo or some other pharmacological intervention for the treatment of Behcet's syndrome. 4. Outcome measures included active ocular inflammatory processes, arthritis, mucocutaneous manifestations (oral ulcer, genital ulcer, erythema nodosum), laboratory changes and major events such as adverse effects and death. DATA COLLECTION AND ANALYSIS: The 32 potentially relevant references were assessed by two independent reviewers (MA, AS) according to the inclusion criteria. Ten trials fit the inclusion criteria and were included in this review. From the 10 included trials, data were independently extracted by the same two observers and crosschecked. The quality of the included trials was assessed independently by two observers (MA, AS) using a validated scale (Jadad 1996). For dichotomous measures, the treatment effect for each trial was calculated using a fixed effect model [Peto model (Petitti 1994)]. The weighted mean differences were based, if available, on end-of-trial results. The analysis was conducted separately for each different intervention. Since the trials could not be pooled it was not possible to carry out a sensitivity analysis by quality scores or a subgroup analysis by drug dosages. Because of this lack of comparability across trials and the small number of trials, we could not conduct a heterogeneity test or a funnel plot. MAIN RESULTS: Ten trials and 679 patients were included. The main results were the lack of efficacy of some of the classic treatments for Behcet's syndrome, including colchicine, cyclophosphamide and steroids for eye involvement, azapropazone and colchicine for arthritis and acyclovir, colchicine and topical interpheron for aphthas. The results confirm the protective effects of cyclosporine and azathioprine for eye involvement and benzathine-penicillin for arthritis. REVIEWER'S CONCLUSIONS: We conclude that further randomized, placebo-controlled, double-blind trials should be carried out to compare cyclosporine, azathioprine and benzathine-penicillin versus placebo in order to make the results generalizable and comparable. PMID- 10796414 TI - Crisis intervention for people with severe mental illnesses. AB - BACKGROUND: A particularly difficult challenge for treatment of people with serious mental illnesses in the community is the delivery of an acceptable level of care during the acute phases of severe mental illness. Crisis intervention models of care were developed as a possible solution to this problem. OBJECTIVES: To review the effects of a crisis intervention model for anyone with serious mental illness experiencing an acute episode, compared to 'standard care'. SEARCH STRATEGY: Relevant randomised trials were identified by searching Biological Abstracts (1985-1998), CINAHL (1982-1998), The Cochrane Library, the Cochrane Schizophrenia Group's Register of trials, EMBASE (1980-1998), MEDLINE (1966 1998), PsycLIT (1974-1998), sociofile (1974-1998) and the ISI database (Science Citations and Social Science Citations). Further references were sought from published trials and their authors. SELECTION CRITERIA: All randomised controlled trials of crisis intervention models (however defined) versus standard care for people with severe mental illnesses (however diagnosed). DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed on an intention-to treat basis. Reviewers assumed that people who left the study early or were lost to follow-up had no improvement. Where possible and appropriate odds ratios (OR) and their 95% confidence intervals (CI) were calculated. The number needed to treat (NNT) was estimated. For continuous data Weighted Mean Differences (WMD) were calculated. Data were inspected for heterogeneity. MAIN RESULTS: None of the five included studies investigated 'crisis intervention' in a pure form. They all used a form of home care for acutely ill people, which included elements of crisis intervention. Despite its ethos, 45% of the home care group were unable to avoid hospital admission during their treatment period. Home care, however, was slightly superior in avoiding repeat admissions (OR 0.63 CI 0.42 - 0. 94), although this result is not robust due to significant heterogeneity. Other results suggest home care reduces loss to follow-up at six and 12 months (OR 0.62, CI 0.42-0.91, NNT 12, CI 6-53; OR 0.65, CI 0.44-0.96, NNT 13, CI 7-130 respectively), reduces family burden (OR 0.20, CI 0.10-0.42, NNT 3, CI 1-5), and is a more satisfactory form of care for both patients and families. No differences in loss, death or mental state were found suggesting home care is as effective as hospital care for these outcomes. All studies found home care to be more cost effective than hospital care but all data were either skewed or unusable. No data on staff satisfaction, carer input, compliance with medication and number of relapses were available. REVIEWER'S CONCLUSIONS: It is difficult to draw any robust conclusions from the data presented in this review. It is also difficult to comment on the effectiveness of crisis intervention in its pure form (i.e. specific results for the acute phase of mental illness only). Overall the review suggests that home care crisis treatment, coupled with an ongoing home care package, is a viable and acceptable way of treating people with serious mental illnesses. Other reviews of more clinically effective home care packages have had similar but more robust results and this review can give tentative support to their findings ( approximately approximately Marshall 1999 approximately approximately ). However, if a pure form of crisis intervention policy is to be specifically practised or implemented it would be hard to justify this outside of a simple well-designed trial. The reviewers also suggest that issues such as staff satisfaction and burnout would be important outcomes to consider in future research. PMID- 10796415 TI - Assertive community treatment for people with severe mental disorders. AB - BACKGROUND: Assertive Community Treatment (ACT) was developed in the early 1970s as a response to the closing down of psychiatric hospitals. ACT is a team-based approach aiming at keeping ill people in contact with services, reducing hospital admissions and improving outcome, especially social functioning and quality of life. OBJECTIVES: To determine the effectiveness of Assertive Community Treatment (ACT) as an alternative to i. standard community care, ii. traditional hospital based rehabilitation, and iii. case management. For each of the three comparisons the main outcome indices were i. remaining in contact with the psychiatric services, ii. extent of psychiatric hospital admissions, iii. clinical and social outcome and iv. costs. SEARCH STRATEGY: Electronic searches of CINAHL (1982 1997), the Cochrane Schizophrenia Group's Register of trials (1997), EMBASE (1980 1997), MEDLINE (1966-1997), PsycLIT (1974-1997) and SCISEARCH (1997) were undertaken. References of all identified studies were searched for further trial citations. SELECTION CRITERIA: The inclusion criteria were that studies should i. be randomised controlled trials, ii. have compared ACT to standard community care, hospital-based rehabilitation, or case management and iii. have been carried out on people with severe mental disorder the majority of whom were aged from 18 to 65. Studies of ACT were defined as those in which the investigators described the intervention as "Assertive Community Treatment" or one of its synonyms. Studies of ACT as an alternative to hospital admission, hospital diversion programmes, for those in crisis, were excluded. The reliability of the inclusion criteria were evaluated. DATA COLLECTION AND ANALYSIS: Three types of outcome data were available: i. categorical data, ii. numerical data based on counts of real life events (count data) and iii. numerical data collected by standardised instruments (scale data). Categorical data were extracted twice and then cross-checked. Peto Odds Ratios and the number needed to treat (NNT) were calculated. Numerical count data were extracted twice and cross-checked. Count data could not be combined across studies for technical reasons (the data were skewed) but all relevant observations based on count data were reported in the review. Numerical scale data were subject to a quality assessment. The validity of the quality assessment was itself assessed. Numerical scale data of suitable quality were combined using the standardised mean difference statistic where possible, otherwise the data were reported in the text or 'Other data tables' of the review. MAIN RESULTS: ACT versus standard community care Those receiving ACT were more likely to remain in contact with services than people receiving standard community care (OR 0.51, 99%CI 0.37-0.70). People allocated to ACT were less likely to be admitted to hospital than those receiving standard community care (OR 0.59, 99%CI 0.41-0.85) and spent less time in hospital. In terms of clinical and social outcome, significant and robust differences between ACT and standard community care were found on i. accommodation status, ii. employment and iii. patient satisfaction. There were no differences between ACT and control treatments on mental state or social functioning. ACT invariably reduced the cost of hospital care, but did not have a clear cut advantage over standard care when other costs were taken into account. ACT versus hospital-based rehabilitation services Those receiving ACT were no more likely to remain in contact with services than those receiving hospital-based rehabilitation, but confidence intervals for the odds ratio were wide. People getting ACT were significantly less likely to be admitted to hospital than those receiving hospital-based rehabilitation (OR 0.2, 99%CI 0.09-0.46) and spent less time in hospital. Those allocated to ACT were significantly more likely to be living independently (OR (for not living independently) 0.19, 99%CI 0.06-0. (A PMID- 10796416 TI - Moderately early (7-14 days) postnatal corticosteroids for preventing chronic lung disease in preterm infants. AB - BACKGROUND: Corticosteroids have been used late in the neonatal period to treat chronic lung disease (CLD) in preterm babies and early to try to prevent it. CLD is likely to be the result of persisting inflammation in the lung and the use of powerful anti-inflammatory drugs like dexamethasone has some rationale. Early use tends to be associated with increased adverse effects so that studies of moderately early treatment (7-14 days postnatal) might have the dual benefits of fewer side effects and onset of action before chronic inflammation is established. OBJECTIVES: To determine if moderately early (7-14 days) postnatal corticosteroid treatment vs control (placebo or nothing) is of benefit in the prevention and/or treatment of early chronic lung disease in the preterm infant. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, Cochrane Database of Controlled Trials, Medline, hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment from 7-14 days of birth in high risk preterm infants were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality up to 28 days and before discharge, failure to extubate, mortality and / or chronic lung disease at 28 days and 36 weeks, CLD at 28 days and 36 weeks, infection, hyperglycaemia, hypertension, severe retinopathy of prematurity (ROP), pneumothorax, necrotizing enterocolitis (NEC), gastrointestinal bleeding, severe intraventricular hemorrhage (IVH), hypertrophic cardiomyopathy, late rescue with dexamethasone and abnormal neurological examination at follow-up were abstracted and analysed using Revman 4.0.4. MAIN RESULTS: Moderately early steroid treatment (vs placebo or nothing) reduced mortality by 28 days, chronic lung disease at 28 days and 36 weeks, and death or chronic lung disease at 28 days or 36 weeks. Earlier extubation was facilitated. There was no significant effect on the rates of pneumothorax, severe ROP, and NEC. Adverse effects included hypertension, hyperglycaemia, gastrointestinal bleeding, hypertrophic cardiomyopathy and infection. Steroid-treated infants were less likely to need late rescue with dexamethasone. There was only one small study of longterm follow-up and it did not show any increase in adverse neurological outcome. REVIEWER'S CONCLUSIONS: Moderately early corticosteroid therapy (started at 7-14 days) reduces neonatal mortality and CLD, but at the cost of important short term adverse effects. No reliable evidence concerning long term effects is provided by the trials included in this review. In view of the evidence of an important increase in cerebral palsy and other adverse neurodevelopmental outcomes from trials in which postnatal steroid was begun either earlier or later than 7-14 days, there are reasonable grounds for extending this concern to moderately early initiation of steroid therapy. More research is urgently needed, including long term follow-up of survivors included in previous and any future trials, before the benefits and risks of postnatal steroid treatment, including initiation at 7-14 days, can be reliably assessed. PMID- 10796417 TI - Synthetic surfactant for respiratory distress syndrome in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To assess the effect of intratracheal administration of synthetic surfactant in premature newborns with established respiratory distress syndrome (RDS). SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactants; limits: age groups, newborn infant; publication types, clinical trial), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, and journal handsearching in the English language. SELECTION CRITERIA: Randomized controlled trials which compared the effect of synthetic surfactant treatment to routine management in the treatment of preterm infants with respiratory distress syndrome. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcome including the incidence of pneumothorax, pulmonary interstitial emphysema, pulmonary hemorrhage, patent ductus arteriosus, necrotizing enterocolitis, apnea of prematurity, intraventricular hemorrhage (any grade, and severe intraventricular hemorrhage), bronchopulmonary dysplasia, neonatal mortality, bronchopulmonary dysplasia or death, retinopathy of prematurity (any retinopathy, and retinopathy greater than Stage 3), mortality at hospital discharge, mortality to one year of age, and cerebral palsy (any, and moderate/severe cerebral palsy) was excerpted from the report of the clinical trials by the reviewer. Data were analyzed according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Six randomized controlled trials of synthetic surfactant treatment of established respiratory distress syndrome were identified. Five of the studies used Exosurf Neonatal (a synthetic surfactant composed of dipalmitoylphosphatidylcholine, hexadecanol and tyloxapol); one small study utilized a mixture of dipalmitoylphosphatidylcholine (DPPC) and phosphatidylglycerol (PG). Treatment with intratracheal Exosurf Neonatal in premature infants with established respiratory distress syndrome improves pulmonary gas exchange and decreases the requirement for ventilatory support. In individual trials, the use of Exosurf Neonatal resulted in a statistically significant reduction in pneumothorax, patent ductus arteriosus, bronchopulmonary dysplasia (BPD), BPD or death at 28 days, and mortality. Similar results are seen when these large trials of Exosurf Neonatal are analyzed in conjunction with the smaller trial of dry powdered DPPC and phosphatidylglycerol (PG). The meta analysis supports a decrease in the risk of pneumothorax (typical relative risk 0.64, 95% CI 0.55, 0.76, typical risk difference -0.09, 95% CI -0.12,-0.06), a decrease in the risk of pulmonary interstitial emphysema (typical relative risk 0.62, 95% CI 0.54, 0.71, typical risk difference -0.12, 95% CI -0.16, -0.09), a decrease in the risk of patent ductus arteriosus (typical relative risk 0.90, 95% CI 0.84, 0.97; typical risk difference -0.06 95% CI -0.10, -0.02), a decrease in the risk of intraventricular hemorrhage (typical relative risk 0.88, 95% CI 0.77, 0.99; typical risk difference -0.04, 95% CI -0.08, -0.00), a decrease in the risk of bronchopulmonary dysplasia (typical relative risk 0.75, 95% CI 0.61, 0.92; typical risk difference -0.04, 95% CI -0.06, -0.01), a decrease in the risk of neonatal mortality (typical relative risk 0. 73, 95% CI 0.61, 0.88; typical risk difference -0.05, 95% CI -0.07, -0.02), a decrease in the risk of bronchopulmonary dysplasia or death at 28 days (typical relative risk 0.73, 95% CI 0.65, 0.83; typical risk difference -0.06, 95% CI -0.11, -0.05), a decrease in the risk of mortality prior to hospital discharge (typical relative risk 0.79, 95% CI 0.68, 0.92; typical risk difference -0.05, 95% CI -0.07, -0.02) and a decrease in the risk of mortality during the first year of life (typical relative risk 0.80, 95% CI 0.69, 0.94; typical risk difference -0.04, 95% CI -0.07, 0.01). (ABS PMID- 10796418 TI - Interventions for shoulder pain. AB - OBJECTIVES: To review the efficacy of common interventions for shoulder pain. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, Cochrane Controlled Trials Register, Medline, Embase, Cinahl, and Science Citation Index) up to May 1998, and hand searched major textbooks, bibliographies of relevant literature, the fugitive literature, and the subject indices of relevant journals including: American College of Rheumatology;British College of Rheumatologists; the Biennial Conference of the Manipulative Physiotherapy Association of Australia;International Federation of Manual Therapists conference proceedings; British Orthopaedic Association;and American Orthopaedic Association. SELECTION CRITERIA: Each identified study was assessed for possible inclusion by two independent reviewers based on the blinded methods sections. The determinants for inclusion were that the trial include an intervention of interest (non-steroidal anti-inflammatory drugs, intra-articular or subacromial glucocorticosteroid injection, oral glucocorticosteroid treatment, physiotherapy, manipulation under anaesthesia, hydrodilatation, or surgery); that treatment allocation was randomized; and that the outcome assessment was blinded. DATA COLLECTION AND ANALYSIS: Methodological quality was assessed by two independent, blinded reviewers. Data relating to selection criteria, outcome measurement and treatment effect was extracted from the blinded trials. Range of motion scores were entered as degrees of restriction to movement, and all pain and overall effect scores were transformed to 100 point scales. For continuous outcome measures, where standard deviation was not reported it was either calculated from the raw data or converted from standard error of the mean. If neither of these were reported, authors were contacted in an effort to obtain the missing values. Effect sizes were calculated and combined in a pooled analysis if study population, endpoint and intervention were comparable. MAIN RESULTS: Thirty one trials met inclusion criteria. Mean methodological quality score was 16.8 (9.5 - 22) out of possible score of 40. Selection criteria varied widely even for the same diagnostic label. There was no uniformity in outcome measures used and their measurement properties were rarely reported. Effect sizes for individual trials were small (-1.4 to 3. 0). The results of only three studies investigating rotator cuff tendonitis could be pooled. Benefit of subacromial steroid injection over placebo for improving range of abduction (weighted difference between means (WMD) 35 degrees, 95% CI 14 to 55) was the only positive finding. REVIEWER'S CONCLUSIONS: There is little evidence to support or refute the efficacy of common interventions for shoulder pain. As well as, the need for further well designed clinical trials, more research is needed to establish a uniform method of defining shoulder disorders and developing outcome measures which are valid, reliable and responsive in these study populations. PMID- 10796419 TI - Cyclophosphamide for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term effects of cyclophosphamide for the treatment of rheumatoid arthritis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's Register, the Cochrane Controlled Trials Register, Medline and Embase up to and including July 1997. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing oral cyclophosphamide against placebo (or an active drug at a dosage considered to be ineffective) in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: Data abstraction was carried out independently by two reviewers. The same two reviewers using Jadad's scale (Jadad 1995) assessed the methodological quality of the RCTs and CCTs. Rheumatoid arthritis outcome measures were extracted from the publications for baseline and end-of-study. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts. Weighted mean differences (WMDs) were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout. MAIN RESULTS: A total of 70 patients were included in the pooled analysis of two trials, 31 receiving cyclophosphamide. A statistically significant benefit was observed for cyclophosphamide when compared to placebo for tender and swollen joint scores: SMDs were -0.57 and -0.59 respectively. The difference in ESR also favoured the active drug but did not reach statistical significance (-12 mm, 95%CI: -26 to 2.5). One trial reported the number of patients developing new or worse erosions: the OR for cyclophosphamide compared to placebo was 0.17 (95% CI: 0.05 to 0.57). Patients receiving placebo were six times more likely to discontinue treatment because of lack of efficacy than patients receiving cyclophosphamide. Withdrawals from adverse reactions were higher in the cyclophosphamide group (Odds ratio=2.9), although this difference was not statistically significant. Side effects from cyclophosphamide included hemorrhagic cystitis, nausea, vomiting, leucopenia, thrombocytopenia, alopecia, amenorrhea and herpes zoster infections. REVIEWER'S CONCLUSIONS: Cyclophosphamide appears to have a clinically and statistically significant benefit on the disease activity of patients with RA, similar to some disease modifying antirheumatic drugs (DMARDs) such as antimalarials or sulfasalazine, but lower than methotrexate. Toxicity however is severe, limiting its use given the low benefit-risk ratio compared to other antirheumatic agents. PMID- 10796420 TI - Moderate-term, low-dose corticosteroids for rheumatoid arthritis. AB - OBJECTIVES: To perform a systematic review of low-dose corticosteroid efficacy in the moderate term for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We conducted a search in MEDLINE from 1966 to 1998, using the keywords "corticosteroids" and "rheumatoid arthritis". We also handsearched all issues of Arthritis and Rheumatism and the Scandinavian Journal of Rheumatology from their dates of first publication to 1994. Furthermore, we examined all Arthritis and Rheumatism abstracts over the 15 year period preceding 1994. References of all identified studies were searched for relevant trials. Authors of unpublished manuscripts were contacted. SELECTION CRITERIA: Studies were selected by two independent reviewers (LC, KS) using a set of predetermined criteria. Specifically, we required that trials be randomized or cross-over and report at least one of the following outcome measures in a quantitative manner: joint tenderness, joint swelling, grip strength, or erythrocyte sedimentation rate (ESR). We also required that trials be of at least three months duration and use prednisone (or a comparable corticosteroid preparation) at a mean dosage of less than or equal to 15 mg/day. We included studies that used either placebo or active drug controls (i.e., comparative studies). DATA COLLECTION AND ANALYSIS: We compared the effectiveness of prednisone to placebo and/or active controls using a fixed effects model for continuous data. A chi square test for homogeneity was performed, and where heterogeneity existed a random effects model was used. We reported results for all available outcomes recommended by the Outcome Measures for Rheumatology Trials (OMERACT) group. These included the number of tender and swollen joints, pain, functional status and ESR. Grip strength was also evaluated. Standardized mean differences (SMD) were used for outcomes assessing the same concept with different scales (eg. swollen joint counts). MAIN RESULTS: Very few studies directly assessed the effectiveness of corticosteroids for RA treatment and many were of poor methodologic quality. Only seven of 34 studies identified by our search met criteria for inclusion. Our results indicated that corticosteroids were significantly more effective than placebo controls for four of six outcomes assessed [standardized mean difference for tender joints = -0.37 (95%CI: -0. 59, -0.14), swollen joints = -0.41 (-0.67, 0.16), pain = -0.43 (-0. 74, -0.12), and functional status = -0.57 (-0.92, 0.22)]. The results for grip strength and ESR were not significant [GS = +0.30 ( 0.19, +0.80), weighted mean difference (WMD) for ESR = -7.03 (-18. 06, +4.01)]. The single trial that compared prednisone to aspirin indicated no statistically significant difference between these groups for joint tenderness (0.10 (-0.35, +0.55) and for ESR [0.00 (-11.09, +11.09]. Overall, the four outcomes assessed in the single trial that compared prednisone to chloroquine suggested that the effectiveness of these two agents is similar [SMD for joint tenderness = +0.23 ( 0.30, +0.75), swollen joints = +0.43 (-0.11, +0. 96), functional status = -0.27 ( 0.80, +0.26), and WMD for ESR = -16. 00 (-30.58, -1.42)]. REVIEWER'S CONCLUSIONS: Based on the limited data available, moderate-term prednisone treatment of RA appears to be superior to placebo and comparable to treatment with aspirin or chloroquine in improving several common rheumatoid arthritis disease activity measures. PMID- 10796421 TI - Nerve blocks (subcostal, lateral cutaneous, femoral, triple, psoas) for hip fractures. AB - BACKGROUND: Various nerve blocks using local anaesthetic agents have been used in order to reduce pain after hip fracture. OBJECTIVES: To determine the effects of nerve blocks (inserted either pre-operatively, operatively or post-operatively) as part of the treatment for a hip fracture. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group trials register, MEDLINE, and bibliographies of trial reports were searched. Date of the most recent search: April 1998. SELECTION CRITERIA: Randomised and quasi-randomised trials involving the use of nerve blocks as part of the care of a hip fracture patient. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, by use of an eight item scale, and extracted data. Wherever appropriate, results of outcome measures were pooled. MAIN RESULTS: Six randomised or quasi-randomised trials involving 229 patients were included. One trial related to insertion of a nerve block pre operatively and the remaining five, to peri-operative insertion. Nerve blocks resulted in a reduction of the quantity of parental or oral analgesia administered to control pain from the fracture/operation or during surgery. It was not possible to demonstrate if this reduction in analgesia use was associated with any clinical benefit. REVIEWER'S CONCLUSIONS: Because of the small number of patients included in this review and the differing type of nerve blocks and timing of insertion, it is not possible to determine if nerve blocks confer any benefit when compared with other analgesic methods as part of the treatment of a hip fracture. Further trials with larger numbers of patients and full reporting of clinical outcomes would be justified. PMID- 10796422 TI - Interventions for treating calcaneal fractures. AB - BACKGROUND: Fracture of the calcaneus (os calcis or heel bone) comprises one to two per cent of all fractures. OBJECTIVES: To identify and evaluate randomised trials of treatments for calcaneal fractures. SEARCH STRATEGY: MEDLINE, EMBASE, CINAHL, the Cochrane Controlled Trials Register, and the Cochrane Musculoskeletal Injuries Group Trials Register were searched. We checked reference lists of relevant articles and contacted trialists and experts in the field. Date of the most recent search: October 1998. SELECTION CRITERIA: Randomised and quasi randomised trials comparing interventions for treating patients with calcaneal fractures. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a 12 item scale, and extracted data. Wherever appropriate and possible, results were pooled. MAIN RESULTS: Of the six relevant randomised trials identified, four were included, one excluded and one is ongoing. All four included trials had methodological flaws. Three trials, involving 134 patients, compared open reduction and internal fixation with non-operative management of displaced intra-articular fractures. Pooled results showed no apparent difference in residual pain (24/40 versus 24/42; Peto odds ratio 0.90, 95% confidence interval 0.34 to 2.36), but a lower proportion of the operative group was unable to return to the same work (11/45 versus 23/45; Peto odds ratio 0.30, 95% confidence interval 0.13 to 0.71), and was unable to wear the same shoes as before (12/52 versus 24/54; Peto odds ratio 0.37, 95% confidence interval 0.17 to 0.84). One trial, involving 23 patients, evaluated impulse compression therapy. At one year there was a mean difference of 1.40 pain units on a visual analogue score (scale 0-10) (95% confidence interval 0.02 to 2.82) in favour of the treated group. The impulse compression group had greater subtalar movement (mean difference 14.0 degrees, 95% confidence interval 3.2 to 24.6) at three months. On average, patients in the impulse compression group returned to work three months earlier than those in the control group. REVIEWER'S CONCLUSIONS: Randomised trials of management of calcaneal fractures are few, small and generally of poor quality. Even where there is some evidence of benefit of operative compared with non-operative treatment, it remains unclear whether the possible advantages of surgery are worth its risks. Given this it seems best to wait for the results of one large ongoing trial on open reduction and internal fixation against conservative treatment. One very small trial suggests that impulse compression therapy for intra-articular calcaneal fractures may be beneficial. More large scale, high quality randomised controlled trials are needed to confirm these results, and to test other interventions in the treatment of calcaneal fractures. PMID- 10796423 TI - Rapid versus slow rate of advancement of feedings for promoting growth and preventing necrotizing enterocolitis in parenterally fed low-birth-weight infants. AB - BACKGROUND: Very premature infants fed by gavage are unable to regulate their own enteral intake. Therefore the rate at which feedings are advanced must be determined by caregivers. While advancing feedings too rapidly may increase the risk of necrotizing enterocolitis, advancing feedings too slowly might result in undernutrition or prolonged hospital stay. OBJECTIVES: For low-birth-weight premature infants receiving parenteral fluids, to assess the effect of different rates of advancement of enteral feedings beginning at the same age on measures of feeding tolerance and neonatal outcome. SEARCH STRATEGY: Search strategies included a Medline search and a search of the Oxford Database of Perinatal Trials; additional references were sought in review articles, relevant chapters of textbooks, a previous systematic review, recent American Pediatric Society - Society for Pediatric Research abstracts, personal files, and personal communication. SELECTION CRITERIA: Only randomized or quasi-randomized trials were considered. Trials were included if premature low-birth-weight infants were studied and if the strategies being compared were different rates of advancement of feedings (accomplished by either differences in volume or concentration) with the onset of feedings at the same postnatal age in each group. DATA COLLECTION AND ANALYSIS: The two reviewers reached consensus for inclusion of trials. Data regarding clinical outcomes were extracted and evaluated by the two reviewers independently of each other. Authors were contacted as needed and feasible to clarify or provide missing data. The specific data that were needed were requested in writing. MAIN RESULTS: Among infants randomized to more rapid rates of advancement of feedings, there was an overall reduction in days to full enteral feeding and days to regain birth weight. There was no effect on necrotizing enterocolitis (relative risk = 0.90, 95% confidence interval = 0. 46 1.77). REVIEWER'S CONCLUSIONS: There are suggested advantages of more rapid rates of advancing feedings in premature low-birth-weight infants (shorter time to regain birth weight and shorter time to achieve full feedings). It is unclear whether this strategy should be adopted as routine practice because of limited information regarding safety (broad confidence intervals for the incidence of necrotizing enterocolitis) and the effect on length of hospital stay (broad confidence intervals). Because different birth weight ranges and different rates of advancement were used in each of these studies, the ideal rate of advancement remains unclear, particularly for extremely-low-birth-weight infants. PMID- 10796424 TI - Prophylactic nasal continuous positive airways pressure for preventing morbidity and mortality in very preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine if prophylactic nasal continuous positive airways pressure (CPAP) commenced soon after birth regardless of respiratory status in the very preterm or very low birth weight infant reduces the use of IPPV and the incidence of chronic lung disease (CLD) without adverse effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts, conferences, symposia, proceedings, expert informants, journal hand searching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random patient allocation of very preterm infants < 32weeks gestation and / or < 1500 gms at birth were eligible. Comparison had to be between prophylactic nasal CPAP commencing soon after birth regardless of the respiratory status of the infant compared with "standard" methods of treatment where CPAP or IPPV is used for a defined respiratory condition. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group, including independent assessment of trial quality and extraction of data by each author, were used. Data were analysed using relative risk (RR). MAIN RESULTS: There are no statistically significant differences in any of the outcomes reported in the single eligible study of 82 very low birth weight infants. More infants in the prophylactic nasal CPAP group received IPPV; however, this difference is not statistically significant. There are trends towards increases in the incidence of CLD (undefined) [RR 2.27 (0.77, 6.65)], of death [RR 3.63 (0.42, 31.08)] and of intraventricular hemorrhage [RR 2.18 (0.84, 5.62)] in the treatment group. The study also found a trend towards a reduction in the incidence of necrotizing enterocolitis in the treatment group [RR 0.40 (0.13, 1.21)]. REVIEWER'S CONCLUSIONS: There is currently insufficient information to make recommendations for clinical practice. The single study reviewed showed no strong evidence of benefit in reducing the incidence of IPPV. The tendency for some adverse outcomes to be increased is of concern and more data are needed to clarify this. A multicentre randomized controlled trial comparing prophylactic nasal CPAP with "standard" methods of treatment is needed to clarify its clinical role. PMID- 10796425 TI - Antifibrinolytic therapy for aneurysmal subarachnoid haemorrhage. AB - BACKGROUND: Re-bleeding is an important cause of death and disability in people with aneurysmal subarachnoid haemorrhage. This is probably due to dissolution of the clot by natural fibrinolytic activity. OBJECTIVES: The objective of this review was to assess the effect of antifibrinolytic treatment in patients with aneurysmal subarachnoid haemorrhage. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register and reference lists of articles. We also contacted drug companies. SELECTION CRITERIA: Randomised trials comparing oral or intravenous antifibrinolytic drugs (tranexamic acid, epsilon amino-caproic acid or an equivalent) with control in people with confirmed subarachnoid haemorrhage. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted the data. All five reviewers assessed trial quality. MAIN RESULTS: Eight trials involving 937 patients were included. Based on 579 patients in two trials, antifibrinolytic treatment did not show any benefit for poor outcome (death, vegetative state or severe disability) with an odds ratio of 1.05, 95% confidence interval 0.76 to 1.46. Death from all causes was not significantly influenced by treatment across all eight trials (odds ratio 0.96, 95% confidence interval 0.72 to 1.26). Antifibrinolytic treatment reduced the risk of re-bleeding reported at the end of follow-up, with some heterogeneity between the trials (odds ratio 0.59, 95% confidence interval 0.42 to 0.81). Treatment increased the risk of cerebral ischaemia in four trials (odds ratio 2.03, 95% confidence interval 1.40 to 2.94). Antifibrinolytic treatment showed no effect on the reported rate of hydrocephalus in four trials (odds ratio 1. 05, 95% confidence interval 0.71 to 1.56). REVIEWER'S CONCLUSIONS: Antifibrinolytic treatment does not appear to benefit people with aneurysmal subarachnoid haemorrhage. However, the trials were all done more than 10 years ago. New strategies may counteract the ischaemia-inducing potential of antifibrinolytic treatment and lead to improved outcome. A trial of combined antifibrinolytic and anti-ischaemia treatment is underway. PMID- 10796426 TI - Thienopyridine derivatives (ticlopidine, clopidogrel) versus aspirin for preventing stroke and other serious vascular events in high vascular risk patients. AB - BACKGROUND: The most widely studied and prescribed antiplatelet agent for the prevention of stroke and other serious vascular events among high vascular risk patients is aspirin. Aspirin inhibits platelet activation by inhibiting platelet cyclooxygenase and thromboxane production, and reduces the odds of a serious vascular event by about a quarter. The thienopyridines (ticlopidine and clopidogrel) inhibit platelet activation by a different mechanism to aspirin (blocking the ADP receptor on platelets), and so may be more effective than aspirin. OBJECTIVES: The objective of this review was to determine the effectiveness and safety of thienopyridine derivatives (ticlopidine and clopidogrel) versus aspirin for the prevention of serious vascular events (stroke, myocardial infarction (MI) or vascular death) in patients at high risk of such events, and specifically in patients with a previous TIA or ischaemic stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register (most recent search: March 1999) and the Antithrombotic Trialists' database, and also contacted Sanofi pharmaceutical company. SELECTION CRITERIA: All unconfounded, double blind, randomised trials directly comparing ticlopidine or clopidogrel with aspirin in high vascular risk patients. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. Additional data were sought from the principal investigators of the largest trial. MAIN RESULTS: Four trials involving a total of 22,656 high vascular risk patients were included. The trials were of high quality and comparable. Aspirin was compared with ticlopidine in three trials (3471 patients) and with clopidogrel in one trial (19,185 patients). Allocation to a thienopyridine was associated with a modest, yet statistically significant, reduction in the odds of a serious vascular event (12. 0% vs 13.0%; OR: 0.91, 95% CI: 0.84 to 0.98; 2p = 0.01), corresponding to the avoidance of 11 (95% CI: 2 to 19) serious vascular events per 1000 patients treated for about two years. There was also a reduction in stroke (5.7% vs 6.4%; OR: 0.88, 95% CI: 0.79 to 0.98; 7 [95% CI: 1 to 13] strokes avoided per 1000 patients treated for two years). Compared with aspirin, thienopyridines produced a significant reduction in the odds of gastrointestinal haemorrhage and other upper gastrointestinal upset, but a significant increase in the odds of skin rash and of diarrhoea. However, the increased odds of skin rash and diarrhoea were greater for ticlopidine than for clopidogrel. Allocation to ticlopidine, but not clopidogrel, was associated with a significant increase in the odds of neutropenia (2.3% vs 0.8%; OR: 2.7, 95% CI: 1.5 to 4.8). In the subset of patients with TIA/ischaemic stroke, the results were similar to those for all patients combined. However, since these patients are at particularly high risk of stroke, allocation to a thienopyridine was associated with a larger absolute reduction in stroke (10.4% vs 12.0%; OR: 0.86, 95% CI: 0.75 to 0.97; 16 [95% CI: 3 to 28] strokes avoided per 1000 patients treated for two years). REVIEWER'S CONCLUSIONS: The available randomised evidence shows that the thienopyridine derivatives are modestly but significantly more effective than aspirin in preventing serious vascular events in patients at high risk (and specifically in TIA/ischaemic stroke patients), but there is uncertainty about the size of the additional benefit. The thienopyridines are also associated with less gastrointestinal haemorrhage and other upper gastrointestinal upset than aspirin, but an excess of skin rash and diarrhoea. The risk of skin rash and diarrhoea is greater with ticlopidine than with clopidogrel. Ticlopidine, but not clopidogrel, is associated with an excess of neutropenia and of thrombotic thrombocytopenic purpura. PMID- 10796427 TI - Cooling therapy for acute stroke. AB - BACKGROUND: Recent studies in acute stroke patients have shown an association between body temperature and prognosis. OBJECTIVES: Our objective was to assess the effects of cooling when applied to patients with acute ischaemic stroke or primary intracerebral haemorrhage. SEARCH STRATEGY: We searched the Cochrane Stroke Group's trial register (last searched in March 1999), plus MEDLINE searched up to November 1998 and EMBASE searched from January 1980 to November 1998. We contacted investigators, pharmaceutical companies and manufacturers of cooling equipment in this field. SELECTION CRITERIA: All completed randomised controlled trials or controlled clinical trials, published or unpublished, where cooling therapy (therapy given by physical devices or antipyretic drugs primarily to lower body temperature independently of basal temperature at the beginning of treatment) was applied up to two weeks of an acute ischaemic stroke or primary intracerebral haemorrhage. DATA COLLECTION AND ANALYSIS: Two reviewers independently searched for relevant trials. MAIN RESULTS: No randomised trials or controlled trials were identified; one placebo-controlled trial of metamizol is currently underway. REVIEWER'S CONCLUSIONS: There is currently no evidence from randomised trials to support the routine use of physical or chemical cooling therapy in acute stroke. Since experimental studies showed a neuroprotective effect of hypothermia in cerebral ischaemia, and hypothermia appears to improve the outcome in patients with severe closed head injury, trials with cooling therapy in acute stroke are warranted. PMID- 10796428 TI - Ultrasound therapy for acute ankle sprains. AB - BACKGROUND: Ultrasound is used in the treatment of a wide variety of musculoskeletal disorders. OBJECTIVES: To evaluate the effects of ultrasound therapy in the treatment of acute ankle sprains. SEARCH STRATEGY: MEDLINE and EMBASE up to December 1998 and databases of the Cochrane Rehabilitation and Related Therapies Field and Musculoskeletal Injuries Group were searched. This was supplemented with citation tracking, and contact with colleagues. Date of the most recent search: April 1999. SELECTION CRITERIA: Randomised or pseudo randomised trials were included in the review if the following conditions were met: at least one study group was treated with active ultrasound; the study included patients with acute lateral ankle sprains; and outcome measures included general improvement, pain, swelling, functional disability, or range of motion. Final selection of papers was conducted by two independent reviewers. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality using a standardised checklist, and extracted data. Whenever possible, the results were analysed on an intention-to-treat basis. Fixed odds ratios together with 95% confidence intervals were calculated for dichotomous outcomes, and weighted or, where different scales were used, standardised mean differences together with 95% confidence intervals for continuous outcome measures. Pooling of data was undertaken where there was clinical homogeneity in terms of subjects, treatments, outcomes, and follow-up time points. MAIN RESULTS: The four included trials, involving 514 patients, were only of modest methodological quality. None of the three placebo-controlled trials (sham ultrasound) could demonstrate statistically significant differences between true and sham ultrasound therapy for any outcome measure at seven days of follow-up. The pooled odds ratio for general improvement was 1.19 (Peto fixed effects model, 95% confidence interval 0.73 to 1.93) for the comparison between ultrasound and sham ultrasound. The differences between intervention groups were generally small, between 0 and 6% for most dichotomous outcomes. However, one trial reported relatively large differences for pain free status (20%) and swelling (25%) in favour of ultrasound treatment. REVIEWER'S CONCLUSIONS: The extent and quality of the available evidence for the effects of ultrasound therapy for acute ankle sprains are limited. The results of three placebo-controlled trials do not support the use of ultrasound in the treatment of ankle sprains. The magnitude of most reported treatment effects appeared to be small, and may be of limited clinical importance. As yet, only few trials are available and no conclusions can be made regarding an optimal and adequate dosage schedule for ultrasound therapy, and whether such a schedule would improve on the reported effectiveness of ultrasound for ankle sprains. PMID- 10796429 TI - Bed rest for acute low back pain and sciatica. AB - BACKGROUND: Low back pain is a common reason for consulting a general practitioner, and advice on daily activities constitutes an important part in the primary care management of low back pain. OBJECTIVES: To assess the effects of bed rest for patients with acute low back pain or sciatica. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trial register, Cochrane Controlled Trials Register, MEDLINE, Embase, Sport, Scisearch, and reference lists of relevant articles. We also contacted authors of relevant articles. Date of the most recent searches: December 1998. SELECTION CRITERIA: We included all randomised studies or quasi randomised studies where at least one comparison group of adult patients with acute low back pain with or without radiation of pain below the knee was advised to rest in bed for at least two days and one group was not, or where comparison groups were advised to stay in bed for different lengths of time. The main outcomes of interest were pain, functional status, recovery and return to work. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed the validity of included trials and extracted data. Investigators were contacted to obtain missing information. MAIN RESULTS: Nine trials with a total of 1435 patients were included. Five trials met all four validity criteria and were assessed to have low risk of bias, while four trials were assessed to have moderate to high risk of bias. Four trials compared bed rest with advice to stay active, and overall the results were heterogeneous. Overall results from two high quality studies indicate no difference in pain intensity at three weeks follow-up [Standardized Mean Difference 0.0 (95%CI: -0.3, 0.2)], and a small difference in functional status in favour of staying active [Weighted Mean Difference 3.2 (on a 0-100 scale) (95%CI 0.6, 5.8)]. Two high quality trials reported no differences in pain intensity between two to three days of bed rest and seven days of bed rest. Another two high-quality trials found no differences between bed rest and exercises in pain intensity or functional status. REVIEWER'S CONCLUSIONS: Bed rest compared to acvice to stay active will at best have small effects, and at worst might have small harmful effects on acute LBP. Differences in effects of advice to stay in bed compared with advice to stay active are small for patients with low back pain with or without sciatica. There is not an important difference in the effects of bed rest compared with exercises in the treatment of acute low back pain, or seven days compared with two to three days of bed rest in patients with low back pain of different duration with and without radiating pain. PMID- 10796430 TI - Hip protectors for preventing hip fractures in the elderly. AB - BACKGROUND: Hip fracture in the elderly is usually the result of a simple fall and hip protectors have been advocated as a means to reduce impact and consequences of such falls. OBJECTIVES: To determine if external hip protectors reduce the incidence of hip fractures in elderly persons following a fall. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group trials register, MEDLINE, and reference lists of relevant articles were searched, and identified trialists contacted. Date of the most recent search: August 1998. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing the use of hip protectors with a control group. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a ten item scale, and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, the data are presented graphically. MAIN RESULTS: Five randomised trials involving 1681 participants were included within the review. All studies involved elderly people in nursing homes or residential care, three within the Scandinavian countries, one in Japan and one in the United Kingdom. The two largest studies involving 1409 participants randomised by nursing home or nursing home ward rather than by the individual (cluster randomisation). One study of 141 individuals was primarily a compliance study. Summation of results from four of these studies gave an occurrence of hip fractures of 13/620 (2.1%) for those allocated to wear hip protectors, against 57/920 (6.2%) to those not allocated to wear protectors. However due to the large number of participants allocated by cluster randomisation it was not possible to demonstrate conclusively that this difference between groups was statistically significant. Only one of the 13 hip fractures that occurred in the individuals allocated to wear hip protectors occurred whilst the protector was worn. No significant adverse effects of the hip protectors were reported but compliance, particularly in the long term, was poor. REVIEWER'S CONCLUSIONS: Hip protectors appear to reduce the risk of hip fracture within a selected population at high risk of sustaining a hip fracture. However, this conclusion is based on four trials of low to moderate quality. As two used cluster randomisation, pooling of data was not possible. The generalisability of the results is unknown beyond high-risk populations. Results from six ongoing trials may clarify this situation. Acceptability by users of the protectors remains a problem, due to discomfort and practicality. PMID- 10796431 TI - Carbamazepine for schizophrenia and schizoaffective psychoses. AB - BACKGROUND: A sizeable minority of people with schizophrenia do not have complete remission of symptoms. A variety of adjunctive treatments have been used to treat schizophrenia, carbamazepine being one. OBJECTIVES: To review the effects of carbamazepine and its derivatives for the treatment of schizophrenia and schizoaffective psychoses. SEARCH STRATEGY: Biological Abstracts (1980-1998), The Cochrane Library (Issue 3, 1998), The Cochrane Schizophrenia Group's Register of Trials (August 1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1886 1998) and PSYNDEX (1974-1998) were searched. Citations from included trials were also inspected and relevant companies and authors contacted for additional data. SELECTION CRITERIA: All randomised controlled trials comparing carbamazepine, or compounds of the carbamazepine family, to placebo or no intervention, whether as sole treatment or as an adjunct to antipsychotic medication for the treatment of schizophrenia and/or schizoaffective psychoses. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were extracted independently by at least two reviewers. Dichotomous data were analysed using Peto odds ratio (OR) and the 95% confidence interval (CI) estimated. Where possible the number needed to treat (NNT) or number needed to harm statistics were calculated. MAIN RESULTS: There is no clear effect of carbamazepine as the sole maintenance treatment for those with schizophrenia. Eight trials provided very limited data on the value of carbamazepine as an adjunct to antipsychotics. Studies were small and poorly reported. Currently no published data provides convincing evidence that adjunctive carbamazepine has an effect on global functioning, mental state, side effects or acceptability of treatment. REVIEWER'S CONCLUSIONS: Based on currently available randomised trial-derived evidence, carbamazepine cannot be recommend for routine clinical use for treatment or augmentation of antipsychotic treatment of schizophrenia. However, for those with a past history of being responsive to carbamazepine, or for those with associated EEG abnormalities, a trial of the drug may be warranted. More data is expected from trialists and the results of this review may be changed by their inclusion. At present large, simple well designed and reported trials are justified especially if focusing on those with both schizophrenia and EEG abnormalities or violent episodes and people with schizoaffective disorders. PMID- 10796432 TI - Bisphosphonates for steroid induced osteoporosis. AB - OBJECTIVES: To assess the effects of bisphosphonates for the prevention and treatment of corticosteroid-induced osteoporosis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, Medline up to 1997 and Embase1988-1997), and selected hand searching of reference lists was conducted. Hand searching of scientific abstracts from relevant meetings for the last five years was also done. An electronic search in Current Contents was done for the last six months. The Cochrane Controlled Trials Register (CCTR) will be searched for future updates. All languages were included in the search. For practical reasons only those in English were included, but all languages will be retrieved and translated for future updates. SELECTION CRITERIA: All controlled clinical trials (CCTs) dealing with prevention or treatment of corticosteroid-induced osteoporosis with bisphosphonates of any type and reporting the outcomes of interest were assessed. Trials had to involve adults only, and subjects had to be taking a mean steroid dose of 7.5 mg/day or more. DATA COLLECTION AND ANALYSIS: All data extraction was performed by two independent reviewers. Outcomes of interest included change in bone mineral density (BMD) at the lumbar spine and femoral neck at six and 12 months. If present, data on number of new fractures and withdrawals due to adverse effects were also extracted. All data extraction was performed by two independent reviewers. Both continuous and dichotomous data were analyzed using fixed effects models. When significant heterogeneity was present, a random effects model was used. MAIN RESULTS: A total of 13 trials, including 842 patients are included in this meta-analysis. Results are reported as a weighted mean difference of the percent change in BMD between the treatment and placebo groups, with trials being weighted by the inverse of their variance. The 95% confidence intervals (95% CI) are presented. At the lumbar spine, the weighted mean difference of BMD between the treatment and placebo groups was 4.3% (95% CI 2.7, 5.9). At the femoral neck, the weighted mean difference was 2.1% (95%CI 0. 01, 3.8). Although there was a 24% reduction in odds of spinal fractures [OR 0.76 (95%CI 0.37, 1.53)], this result was not statistically significant. REVIEWER'S CONCLUSIONS: Bisphosphonates are effective at preventing and treating corticosteroid-induced bone loss at the lumbar spine and femoral neck. Efficacy regarding fracture prevention cannot be concluded from this analysis, although bone density changes are correlated with fracture risk. PMID- 10796433 TI - Surgery for lumbar disc prolapse. AB - OBJECTIVES: The primary rationale for surgery for disc prolapse is to relieve nerve root irritation or compression due to herniated disc material. Claims of the merits of alternative surgical procedures are made without clear evidence about clinical outcomes. The objective of this review was to assess the effects of surgical interventions for the treatment of lumbar disc prolapse. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline, Embase, Biosis, Dissertation Abstracts, Index to UK Thesis, and reference lists of the retrieved articles up to March 1997 and we corresponded with experts. SELECTION CRITERIA: Randomised and quasi-randomised trials of the surgical management of lumbar disc prolapse. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data from published papers. Additional information was sought from the authors if necessary. MAIN RESULTS: Twenty-seven trials were found. There were methodological weaknesses in many of the trials. Sixteen of the 27 trials were of some form of chemonucleolysis. Ten trials compared different surgical techniques, although only one of these compared surgical discectomy with conservative management. Surgical discectomy produced better clinical outcomes than chemonucleolysis with chymopapain, and chemonucleolysis produced better clinical outcomes than placebo. Three trials showed no difference in clinical outcomes between microdiscectomy and standard discectomy. Three trials failed to show a significant reduction in scar formation or improved clinical outcomes by inserting an inter-position membra ne to cover the spinal dura after discectomy. Three trials of percutaneous discectomy provided moderate evidence that it produces poorer clinical outcomes than standard discectomy or chymopapain. We found no published randomised trials of laser discectomy. REVIEWER'S CONCLUSIONS: Chemonucleolysis is more effective than placebo and it is less invasive but less effective than surgical disectomy. Surgical discectomy for carefully selected patients with sciatica due to lumbar disc prolapse provides faster relief from the acute attack than conservative management, although any positive or negative effects on the lifetime natural history of the underlying disc disease are unclear. PMID- 10796434 TI - Acupuncture for low back pain. AB - BACKGROUND: Although low back pain is usually a self-limiting and benign disease that tends to improve spontaneously over time, a large variety of therapeutic interventions are available for the treatment of low back pain. OBJECTIVES: The objective of this review was to assess the effects of acupuncture for the treatment of non-specific low back pain. SEARCH STRATEGY: We searched the Cochrane Complementary Medicine Field trials register, the Cochrane Controlled Trials Register (1997, issue 1), Medline (1966 - 1996), Embase (1988 - 1996), Science Citation Index and reference lists of articles. SELECTION CRITERIA: Randomised trials of all types of acupuncture treatment that involves needling for subjects with non-specific low back pain. DATA COLLECTION AND ANALYSIS: Two reviewers blinded with respect to authors, institution and journal independently assessed trial quality and extracted data. MAIN RESULTS: Eleven trials were included. The methodological quality was low. Only two trials were of high quality. Three trials compared acupuncture to no treatment, which were of low methodological quality and provide conflicting evidence. There was moderate evidence from two trials that acupuncture is not more effective than trigger point injection or transcutaneous electrical nerve stimulation (TENS). There was limited evidence from eight trials that acupuncture is not more effective than placebo or sham acupuncture for the treatment of chronic low back pain. REVIEWER'S CONCLUSIONS: The evidence summarised in this systematic review does not indicate that acupuncture is effective for the treatment of back pain. PMID- 10796435 TI - Surgery for degenerative lumbar spondylosis. AB - BACKGROUND: This section is under preparation and will be included in the next issue OBJECTIVES: Degenerative conditions affecting the lumbar spine are variously described as lumbar spondylosis or degenerative disc disease (which we regarded as one entity) and are associated with back pain, instability, spinal stenosis and degenerative spondylolisthesis. The objective of this review was to assess the effects of surgical interventions for the treatment of degenerative lumbar spondylosis. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline, Embase, Biosis, Dissertation Abstracts, Index to UK Thesis, and reference lists of the retrieved articles and we corresponded with experts. SELECTION CRITERIA: Randomised or quasi-randomised trials of surgical treatment of lumbar spondylosis DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data from published papers. Additional information was sought from the authors if necessary. MAIN RESULTS: Fourteen published trials of all forms of surgical treatment for degenerative lumbar spondylosis were identified. There were many serious weaknesses of trial design, including poor methods of randomisation, lack of blinding and lack of independent assessment of outcome which at times gave considerable potential for bias. Most of the published results were reporting on technical surgical outcomes with some crude ratings of clinical outcome, but few patient-centred outcomes of pain, disability or capacity for work. There was a particular lack of long-term outcomes. This review found no published trials comparing any form of surgery for degenerative lumbar spondylosis compared with natural history, placebo, or any form of conservative treatment. Nine trials randomly compared instrumented and non instrumented fusion. Instrumented fusion produced a higher fusion rate (though that needs to be qualified by the difficulty of assessing fusion in the presence of metal-work) but did not improve clinical outcomes and there is evidence that it may be associated with higher complication rates. The few and heterogeneous trials on spondylolisthesis, spinal stenosis and nerve compression permitted very limited conclusions. REVIEWER'S CONCLUSIONS: There is no scientific evidence about the effectiveness of any form of surgical decompression or fusion for degenerative lumbar spondylosis compared with natural history, placebo, or conservative treatment. PMID- 10796436 TI - Surgical treatment for meniscal injuries of the knee in adults. AB - BACKGROUND: Injuries to the knee menisci are common and operations to treat them are among the most common procedures performed by orthopaedic surgeons. OBJECTIVES: To evaluate the effects of common surgical interventions in the treatment of meniscal injuries of the knee. The four comparisons under test were: a) surgery versus conservative treatment, b) partial versus total meniscectomy, c) excision versus repair of meniscal tears, d) surgical access, in particular arthroscopic versus open. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, MEDLINE and bibliographies of published papers. Date of the most recent search: August 1998. SELECTION CRITERIA: All randomised and quasi-randomised trials which involved the above four comparisons or which compared other surgical interventions for the treatment of meniscal injury. DATA COLLECTION AND ANALYSIS: Trial inclusion was agreed by both reviewers who independently assessed trial quality, by use of a 12 item scale, and extracted data. Where possible and appropriate, data were presented graphically. MAIN RESULTS: Three trials, involving 260 patients, which addressed two (partial versus total meniscectomy; surgical access) comparisons were included. Partial meniscectomy may allow a slightly enhanced recovery rate as well as a potentially improved overall functional outcome including better knee stability in the long term. It is probably associated with a shorter operating time with no apparent difference in early complications or re-operation between partial and total meniscectomy. The long term advantage of partial meniscectomy indicated by the absence of symptoms (symptoms or further operation at six years or over: 14/98 versus 22/94; Peto odds ratio 0.55, 95% confidence interval 0.27 to 1.14) or radiographical outcome was not established. The results available from the only trial comparing arthroscopic with open meniscectomy were very limited in terms of patient numbers and length of follow-up. However it is likely that partial meniscectomy via arthroscopy is associated with shorter operating times and a quicker recovery. REVIEWER'S CONCLUSIONS: The lack of randomised trials means that no conclusions can be drawn on the issue of surgical versus non surgical treatment of meniscal injuries, nor meniscal tear repair versus excision. In randomised trials so far reported, there is no evidence of difference in radiological or long term clinical outcomes between arthroscopic and open meniscal surgery, or between total and partial meniscectomy. Partial meniscectomy seems preferable to the total removal of the meniscus in terms of recovery and overall functional outcome in the short term. PMID- 10796437 TI - Individual psychodynamic psychotherapy and psychoanalysis for schizophrenia and severe mental illness. AB - BACKGROUND: People with schizophrenia and severe mental illness may need much support from health care professionals, in most cases over a long period of time. Research on the effects of psychotherapy for schizophrenia shows mixed results. Although pharmacological interventions remain the treatment of choice for schizophrenia patients, it is also of interest to look at the effects of treatment methods focusing on psychosocial factors affecting schizophrenia. OBJECTIVES: To review the effects of individual psychodynamic psychotherapy and/or psychoanalysis for people with schizophrenia or severe mental illness. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1985-1999), CINAHL (1982-1999), The Cochrane Library CENTRAL (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (1999), Dissertation Abstracts On disc (1866 1999), EMBASE (1980-1999), MEDLINE (1966-1999), PsycLIT (1974-1999), and Sociofile (1974-1998) were made. Authors of included trials were contacted for information on further trials. SELECTION CRITERIA: All randomised trials of individual psychodynamic psychotherapy or psychoanalysis for people with schizophrenia or severe mental illness (however defined) were selected. DATA COLLECTION AND ANALYSIS: Data were independently extracted by at least two reviewers. For dichotomous data relative risks (RR) were calculated and for continuous data weighted mean differences (WMD) between groups were calculated. MAIN RESULTS: No trials of a psychoanalytic approach were identified. Data are sparse for all comparisons involving a psychodynamic approach. There is no evidence of any positive effect of psychodynamic therapy and the possibility of adverse effects seems never to have been considered. The psychodynamic approach may be more acceptable to people than a more cognitive reality-adaptive therapy. REVIEWER'S CONCLUSIONS: Current data do not support the use of psychodynamic psychotherapy techniques for hospitalised people with schizophrenia. If psychoanalytic therapy is being used for people with schizophrenia there is an urgent need for trials. PMID- 10796438 TI - Depot haloperidol decanoate for schizophrenia. AB - BACKGROUND: The mainstay of treatment for schizophrenia is the antipsychotic group of drugs. These are usually given orally but compliance with medication given by this route may be difficult to quantify. Problems with treatment adherence are common. The development of depot injections in the 1960s gave rise to their extensive use as a means of long-term maintenance treatment. Haloperidol decanoate is one depot drug available in clinical practice. OBJECTIVES: To assess the effects of haloperidol decanoate versus oral anti-psychotics and other depot antipsychotic preparations for people with schizophrenia in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Relevant trials were identified by searching Biological Abstracts (1982-1998), Cochrane Library (Issue 2, 1998), Cochrane Schizophrenia Group's Register (June 1998), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1974-1998). References of all identified trials were also inspected for more studies. SELECTION CRITERIA: All relevant randomised trials focusing on people with schizophrenia where haloperidol decanoate, oral anti-psychotics or other depot preparations were compared. Outcomes such as death, clinically significant change in global function, mental state, relapse, hospital admission, adverse effects and acceptability of treatment were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data Mantel-Haenszel odds ratios (OR) with the 95% confidence intervals (CI) were estimated. Where possible, the number needed to treat statistic (NNT) was calculated. Analysis was by intention-to-treat. Normal continuous data were summated using the weighted mean difference (WMD). Scale data were presented only for those tools that had attained pre-specified levels of quality. MAIN RESULTS: In a haloperidol decanoate versus placebo comparison, two small studies reported that significantly fewer people on depot left early (OR 0.09 CI 0.03-0.21, NNT 2 CI 1-3) or experienced no important improvement in mental state (OR 0. 04 CI 0.01-0.15). Zississ (1982) suggested that those taking haloperidol decanoate would need less additional antipsychotic medication (OR 0.14 Cl 0.04-0.55, NNT 2 CI 1-5). Haloperidol decanoate was compared to oral haloperidol in a single trial that showed no differences in global impression, mental state or side effects ( approximately approximately Zuardi 1983 approximately approximately , n=22). Compliance with medication was not reported in this study. Eight trials compared haloperidol decanoate to other depot neuroleptics and again no differences were found for the outcomes of death, global impression, mental state, behaviour, or side effects. REVIEWER'S CONCLUSIONS: Haloperidol decanoate may have a substantial effect in improving the symptoms and behaviour associated with schizophrenia in comparison to placebo, but data are remarkably sparse. There are no discernible differences between the depot form of haloperidol and its oral equivalent. For those needing and willing to take the drug, the means of administration is then a matter of individual choice and clinical judgement. As there are no clear differences between haloperidol decanoate and other depots, the choice of depot medication could also be individually tailored and patient preference exercised. Well-conducted and reported randomised trials are needed comparing haloperidol decanoate with other depots but the comparison of haloperidol decanoate to oral antipsychotics is a priority. PMID- 10796439 TI - Venepuncture versus heel lance for blood sampling in term neonates. AB - BACKGROUND: Heel lance has been the conventional method of blood sampling in neonates for screening tests. Neonates undergoing this procedure experience pain. Despite various studies evaluating the role of pharmacological and non pharmacological interventions to date, there are no effective and practical methods to alleviate pain from heel lance. OBJECTIVES: To determine whether venepuncture or heel lance is less painful and more effective for blood sampling in term neonates. SEARCH STRATEGY: Systematic search in accordance with Cochrane Neonatal Collaborative Review Group. All randomized controlled trials which compared pain response to venepuncture vs. heel lance were searched using MEDLINE, Embase, CINAHL, the Cochrane Library, reference lists of identified trials and personal files. No language restrictions were applied. SELECTION CRITERIA: Randomized controlled trials which compared pain response to venepuncture vs. heel lance were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding the primary outcome of pain response to venepuncture vs. heel lance as assessed by validated pain measures were abstracted. Secondary outcomes including the need of repeat blood sampling, bruising/hematoma at local site, and parental perception of their own anxiety and infant's pain were also abstracted. All data were analysed using Revman 3.1. When possible, meta-analysis was done using relative risk (RR) and event rate difference (ERD), along with their 95% confidence intervals (CI). If ERD was significant, number needed to treat (NNT) was calculated. MAIN RESULTS: Two trials were eligible for inclusion in the review. One study using Neonatal Infant Pain Scale (NIPS) and one study using Neonatal Facial Action Coding System (NFCS) and cry duration showed statistically significantly lower pain scores for venepuncture as compared to heel lance. In one study, maternal anxiety was noted to be higher in the venepuncture group as compared to heel lance group prior to the procedure; however, after observing the procedure, mothers rated their infant's pain to be lower in the venepuncture group. A meta-analysis was performed on the need for at least one additional skin puncture to obtain the required amount of blood using venepuncture as compared to heel lance. The relative risk for requiring more than one skin puncture for venepuncture versus heel lance was 0.19 [95% confidence interval (CI) 0.10, 0.38]. The risk difference was -53% (95% CI -66%, -40%). The number needed to treat (NNT) to avoid one repeat skin puncture was 1.9 (95% CI 1.5, 2.5). REVIEWER'S CONCLUSIONS: Venepuncture, when performed by a trained phlebotomist, appears to be the method of choice for blood sampling in term neonates. For each two venepunctures instead of heel lance, the need for one additional skin puncture can be avoided. In view of the promising results derived from small studies with some methodological limitations, further well designed randomized controlled trials need to be conducted. The interventions should be compared in settings where several individuals perform the venepuncture and/or the heel lance. PMID- 10796440 TI - Penicillamine for rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term effects of D-penicillamine for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, Medline up to and including December 1998 and Embase from 1988-1998. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials and controlled clinical trials comparing D-penicillamine against placebo in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: The methodological quality of the trials was assessed independently by two reviewers (CS, EB) and checked by a third (MS) using a validated quality assessment tool. Rheumatoid arthritis outcome measures were extracted from the publications for the six-month endpoint and stratified according to D-penicillamine dosages: low (<500mg/day), moderate (500 to <1000mg/day) and high (1000 mg/day or greater). Data was abstracted by one reviewer and checked by a second (CS, MS). The pooled analysis was performed using the standardized mean difference for joint counts, pain and global assessments. The weighted mean difference was used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals and adverse reactions. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout, since no statistical heterogeneity was found. MAIN RESULTS: Six trials were identified, with 425 patients randomized to D-penacillamine and 258 to placebo. A statistically significant benefit was observed for D-penicillamine when compared to placebo for all three-dose ranges and for most outcome measures including: tender joint counts, pain, physician's global assessments and ESR. The standardized weighted mean differences between treatment and placebo in moderate doses were -0. 51 [95% CI -0.88, -0.14] for tender joint counts, -0.56 (95% CI 0. 87, -0.26) for pain and -0.97 (95% CI -1.25, -0.70) for global assessment. The difference for ESR was -10.6 mm/hr. Similar results were observed for the higher dose group. Total withdrawals were significantly higher in the moderate and high dosage D-penicillamine groups (OR=1.63 and 2.13 respectively), mostly due to increased adverse reactions (OR = 2.60 and 4.95 respectively), including renal and hematological abnormalities. REVIEWER'S CONCLUSIONS: D-penicillamine appears to have a clinically and statistically significant benefit on the disease activity of patients with rheumatoid arthritis. Its efficacy appears to be similar to that of other disease modifying anti-rheumatic drugs (DMARDs), but with a significantly higher toxicity. Its effects on long-term functional status and radiological progression are not clear from this review. PMID- 10796441 TI - Azathioprine for rheumatoid arthritis. AB - OBJECTIVES: To assess the short-term effects of azathioprine for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, Medline up to and including July 1998 and Embase from 1988-1998. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials and controlled clinical trials comparing azathioprine against placebo in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: Data was extracted independently by two reviewers (CS, EB); disagreements were resolved by discussion or third party adjudication (MS). The same reviewers (CS, EB) assessed the methodological quality of the trials using a validated quality assessment tool. Rheumatoid arthritis outcome measures were extracted from the publications for the six-month endpoint. The pooled analysis was performed using standardized mean differences for joint counts, pain and functional status assessments. Weighted mean differences were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals and for adverse reactions. The 95% confidence intervals (95% CI) are presented. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout, since no statistical heterogeneity was found. MAIN RESULTS: Three trials with a total of 81 patients were included in the analysis. Forty patients were randomized to azathioprine and forty-one to placebo. A pooled estimate was calculated for two outcomes. A statistically significant benefit was observed for azathioprine when compared to placebo for tender joint scores. The standardized weighted mean difference between treatment and placebo was -0.98 (95% CI -1.45, 0.50). Withdrawals from adverse reactions were significantly higher in the azathioprine group OR=4.56 (95% CI 1.16, 17.85). REVIEWER'S CONCLUSIONS: Azathioprine appears to have a statistically significant benefit on the disease activity in joints of patients with RA. This evidence however is based on a small number of patients, included in older trials. Its effects on long-term functional status and radiological progression were not assessed due to lack of data. Toxicity is shown to be higher and more serious than that observed with other disease-modifying anti-rheumatic drugs (DMARDs). Given this high risk to benefit ratio, there is no evidence to recommend the use of azathioprine over other DMARDs. PMID- 10796442 TI - Depot flupenthixol decanoate for schizophrenia or other similar psychotic disorders. AB - BACKGROUND: Anti-psychotic drugs are the mainstay treatment for schizophrenia and similar psychotic disorders. Long-acting depot injections of drugs such as flupenthixol decanoate are extensively used as a means of long-term maintenance treatment. OBJECTIVES: To evaluate the effects flupenthixol decanoate in comparison with placebo, oral antipsychotics and other depot neuroleptic preparations for people with schizophrenia and other severe mental illnesses, in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Relevant trials were identified by searching Biological Abstracts (1982-1998), Cochrane Library (Issue 2, 1998), Cochrane Schizophrenia Group's Register (December 1998), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1974-1998). The references of all identified trials were inspected for more studies and the first author of each included trial and relevant pharmaceutical companies were contacted. SELECTION CRITERIA: All randomised controlled trials that focused on people with schizophrenia or other similar psychotic disorders where flupenthixol decanoate had been compared to placebo or other antipsychotic drugs. All clinically relevant outcomes were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data Peto odds ratios (OR) with 95% confidence intervals (CI) were estimated. Where possible, the number needed to treat statistic (NNT) was also calculated. Analysis was by intention-to-treat. Normal continuous data were summated using the weighted mean difference (WMD). Scale data were presented only for those tools that had attained pre-specified levels of quality. MAIN RESULTS: No trials compared flupenthixol decanoate to placebo. One small study compared flupenthixol decanoate with an oral antipsychotic (penfluridol). There were no clear differences between the two preparations. When flupenthixol decanoate was compared to other depot preparations, there were no differences between depots for outcomes such as death, global impression, relapse (OR 1.16 CI 0.7-1.9) or leaving the study early (OR 1.00 CI 0.6-1.7). Two small studies suggest that flupenthixol decanoate is responsible for less movement disorders than other depot antipsychotic drugs (OR 0.23 CI 0.08-0.7, NNT 5). This finding did not hold for specific side effects, such as tremor (OR 1.2 CI 0.3-4) and tardive dyskinesia (OR 1.60 CI 0.4-6). Two trials comparing high doses of flupenthixol decanoate to the standard approximately 40mg per injection reported no significant difference for the outcome of relapse (OR 0.32 CI 0.09-1.2). One small (n=59) trial comparing a very low dose of flupenthixol decanoate ( approximately 6 mg/IM) to a very low dose approximately 9 mg per injection also reported no difference in relapse rates (OR 0.3 CI 0.1-1.1). REVIEWER'S CONCLUSIONS: From the data reported in clinical trials, it would be understandable if those suffering from schizophrenia, who are willing to take flupenthixol decanoate, would request the standard dose rather than the high dose. In the current state of evidence, there is nothing to choose between flupenthixol decanoate and other depot antipsychotics. The choice of which depot to use must therefore be based on clinical judgement and the preferences of people with schizophrenia and their carers. Managers and policy makers should expect better data than the research community has provided thus far. This review highlighted the need for large, well-designed and reported randomised clinical trials to address the effects of flupenthixol decanoate, in particular when compared to oral antipsychotics. Future studies should also consider hospital and service outcomes, satisfaction with care and record economic data. PMID- 10796443 TI - Postnatal phenobarbitone for the prevention of intraventricular hemorrhage in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: To determine whether postnatal administration of phenobarbitone to preterm infants reduces the risk of intraventricular hemorrhage (IVH), neurodevelopmental impairment or death. SEARCH STRATEGY: See the Search Strategy of the Neonatal Collaborative Review Group. The reviewer has been a active trialist in this area and has personal contact with many groups in this field. Journals handsearched from 1976 (when cranial CT scanning started) to November 1998 include: Pediatrics, J Pediatrics, Archives of Disease in Childhood, Pediatric Research, Developmental Medicine and Child Neurology, Acta Paediatrica, European J of Pediatrics, Neuropediatrics, New England J of Medicine, Lancet and British Medical J. The National Library of Medicine (USA) database (via PubMed) and the Cochrane Controlled Trials Register were searched through to November 1998 using the MeSH terms intraventricular hemorrhage, newborn infants, premature infant, intracranial hemorrhage, phenobarbitone, phenobarbital. The searches were not limited to the English language, as long as the article included an English abstract. Promising articles were read in the original language or translated. SELECTION CRITERIA: Included were randomized or quasi-randomized controlled trials in which phenobarbitone was given to preterm infants identified as being at risk of IVH because of gestational age below 34 weeks, birthweight below 1500 g, or respiratory failure. Adequate determination of IVH by ultrasound or CT was also required. DATA COLLECTION AND ANALYSIS: In addition to details of patient selection and control of bias, the details of the administration of phenobarbitone were extracted. The end-points searched for included: IVH ( with grading), posthemorrhagic ventricular dilatation or hydrocephalus, neurodevelopmental impairment and death. In addition, possible adverse effects of phenobarbitone such as hypotension, mechanical ventilation, pneumothorax, hypercapnia, and acidosis were searched for. MAIN RESULTS: Nine controlled trials were included with 740 infants recruited. There was heterogeneity between trials for the outcome IVH, with one trial finding a significant decrease in IVH and another trial finding an increase in IVH in the group receiving phenobarbitone. Meta-analysis showed no difference between the phenobarbitone treated group and the control group in either IVH (typical relative risk 1.04, CI 0.87, 1.25), severe IVH (typical relative risk 0.91, CI 0.66, 1.27), posthemorrhagic ventricular dilatation (typical relative risk 0.89, CI 0.38, 2.08), severe neurodevelopmental impairment (typical relative risk 1.44, CI 0.41, 5.04) or death before hospital discharge (typical relative risk 0.88, CI 0.64, 1.21) There was a consistent trend in the trials towards increased use of mechanical ventilation in the phenobarbitone treated group, which was supported by the meta-analysis (typical relative risk 1.18, CI 1.06, 1.32; typical risk difference 0.129, CI 0.045, 0.213), but there was no significant difference in pneumothorax, acidosis or hypercapnia. REVIEWER'S CONCLUSIONS: Postnatal administration of phenobarbitone cannot be recommended as prophylaxis to prevent IVH in preterm infants and is associated with an increased need for mechanical ventilation. PMID- 10796444 TI - Peripheral retinal ablation for threshold retinopathy of prematurity in preterm infants. AB - BACKGROUND: This section is under preparation and will be included in the next issue. OBJECTIVES: In premature infants with threshold retinopathy of prematurity (ROP) does peripheral retinal ablation, by any means, reduce the incidence of adverse ophthalmic outcome? SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included a search of the Cochrane Neonatal Group Register of Clinical Trials, MEDLINE, EMBASE, previous reviews including cross references, abstracts from pediatric and ophthalmologic meetings, letters and expert informants. Search terms included "Retinopathy of Prematurity" [MeSH Terms], "Retrolental Fibroplasia" [All Fields] and "Lightcoagulation" [All Fields] or "Cryosurgery" [All Fields]. In addition, a personal bibliographic database was used as a cross-reference. SELECTION CRITERIA: All trials in human premature infants with threshold ROP utilizing random or quasi random allocation to either peripheral retinal ablation of the avascular retina, by any means, or concurrent control group with independent outcome assessment were initially selected for review. Following methodologic review, only studies using random allocation were selected for data extraction. DATA COLLECTION AND ANALYSIS: Relevance and validity were assessed by the two authors and consensus reached. Each author extracted clinical outcomes from valid reports independently. Data analysis was conducted according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Two randomised trials were identified. Data from these studies show that peripheral retinal ablation reduces the risk of (1) early unfavorable retinal structure from 47. 9% to 28.1% (absolute risk reduction 19.8% [95% CI 27.9 - 11.8%]), (2) unfavorable retinal structure in early childhood from 44.3% to 26.3% (absolute risk reduction 18% [95% CI 27.0 - 9.1%]) and (3) unfavorable visual acuity in early childhood from 63% to 50.6% (absolute risk reduction 12.2% [95% CI 21.2 - 3.1]). In addition, visual fields in sighted eyes were slightly smaller in the treated (51.3 degrees +/- 11.8 degrees ) group as compared to the control (58.2 degrees +/- 14.5 degrees ) group. REVIEWER'S CONCLUSIONS: Peripheral retinal ablation reduces the incidence of adverse ophthalmic outcome in premature infants with threshold ROP. In sighted eyes, peripheral retinal ablation may reduce the size of the visual field. At this stage, long term outcomes remain unknown. PMID- 10796445 TI - Depot perphenazine decanoate and enanthate for schizophrenia. AB - BACKGROUND: Anti-psychotic drugs are usually given orally but compliance with medication given by this route may be difficult to quantify. The development of depot injections in the 1960s gave rise to extensive use of depots as a means of long-term maintenance treatment. Perphenazine decanoate and enanthate are depot antipsychotics that belong to the phenothiazine family and have a piperazine ethanol side chain. OBJECTIVES: To assess the effects of depot perphenazine decanoate and enanthate versus placebo, oral anti-psychotics and other depot antipsychotic preparations for people with schizophrenia in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Biological Abstracts (1982-1998), the Cochrane Library (Issue 2, 1998), the Cochrane Schizophrenia Group's Register (June 1998), EMBASE (1980-1998), MEDLINE (1966-1998), and PsycLIT (1974-1998) were searched. References of all identified trials were also inspected for more studies and industry contacted. SELECTION CRITERIA: Randomised clinical trials focusing on people with schizophrenia where depot perphenazine decanoate and enanthate, oral anti-psychotics or other depot preparations were compared. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data Peto odds ratios (OR) with the 95% confidence intervals (CI) were estimated. Where possible, the number needed to treat statistic (NNT) was calculated. Analysis was by intention-to-treat. MAIN RESULTS: One study of six months duration, compared perphenazine enanthate to clopenthixol decanoate. There was no differences between the two for outcomes of global improvement, relapse and leaving the study early. More people in the perphenazine enanthate group required anticholinergic drugs than those allocated to clopenthixol decanoate (OR 3.6 CI 1.2-10, NNT 10). A single study (n=64, duration six weeks) compared perphenazine enanthate and its longer acting decanoate ester. Data on relapse and leaving the study early failed to show convincing differences. The enanthate group, however, experienced more movement disorders (OR 0.2 CI 0.06-0.7) than those allocated the decanoate ester of the same drug (NNT 4.0) and required more anticholinergic drugs (OR 0.2 CI 0.08-0.7, NNT 3.7). REVIEWER'S CONCLUSIONS: Depot perphenazine is in clinical use in the Nordic countries, Belgium, Portugal and the Netherlands. At a conservative estimate a quarter of a million people suffer from schizophrenia in those countries and could be treated with depot perphenazine. The total number of participants in the two trials with useful data is 236. Neither study observes the effect of oral versus depot antipsychotic drugs. Until well conducted and reported randomised trials are undertaken clinicians will be in doubt as to the effects of perphenazine depots and people with schizophrenia should exercise their own judgement or ask to be randomised. PMID- 10796446 TI - Depot fluspirilene for schizophrenia. AB - BACKGROUND: Anti-psychotic drugs are the mainstay treatment for schizophrenia and similar psychotic disorders. Long-acting depot injections of drugs such as fluspirilene are extensively used as a means of long-term maintenance treatment. OBJECTIVES: To assess the effects of depot fluspirilene versus placebo, oral anti psychotics and other depot antipsychotic preparations for people with schizophrenia in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Relevant trials were identified by searching Biological Abstracts (1982 1998), Cochrane Library (Issue 2, 1998), Cochrane Schizophrenia Group's Register (June 1998), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1974-1998). References of all identified trials were also inspected for more studies. SELECTION CRITERIA: All relevant randomised trials focusing on people with schizophrenia where depot fluspirilene, oral anti-psychotics or other depot preparations were compared. Outcomes such as death, clinically significant change in global function, mental state, relapse, hospital admission, adverse effects and acceptability of treatment were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data, Peto odds ratios (OR) with the 95% confidence intervals (CI) were estimated. Where possible, the number needed to treat statistic (NNT) was calculated. Analysis was by intention-to-treat. Normal continuous data were summated using the weighted mean difference (WMD). Scale data were presented only for those tools that had attained pre-specified levels of quality. MAIN RESULTS: Seven studies were included. Most comparisons included very few participants. There are no convincing data showing fluspirilene decanoate's advantage over oral chlorpromazine or other depot antipsychotics. No study reported on hospital and service outcomes or commented on participants' overall satisfaction with care. Economic outcomes were not recorded by any of the included studies. REVIEWER'S CONCLUSIONS: The total numbers in each comparison were small and there were no clear differences demonstrated between fluspirilene and oral medication or other depots. The choice of whether to use fluspirilene as a depot medication and whether it has advantages over other depots, cannot, at present, be informed by trial-derived data. Well-conducted and reported randomised trials are still needed to inform practice. PMID- 10796447 TI - Depot bromperidol decanoate for schizophrenia. AB - BACKGROUND: Anti-psychotic drugs are the mainstay treatment for schizophrenia. Long-acting depot injections of drugs such as bromperidol decanoate are extensively used as a means of long-term maintenance treatment. OBJECTIVES: To assess the effects of depot bromperidol versus placebo, oral anti-psychotics and other depot antipsychotic preparations for people with schizophrenia in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Relevant trials were identified by searching Biological Abstracts (1982-1999), Cochrane Library (Issue 2, 1999), Cochrane Schizophrenia Group's Register (May 1999), EMBASE (1980-1999), MEDLINE (1966-1999) and PsycLIT (1974-1999). References of all identified trials were inspected and Janssen Cilag contacted in order to identify more trials. SELECTION CRITERIA: All randomised trials focusing on people with schizophrenia where depots bromperidol, oral anti-psychotics or other depot preparations were sought. Primary outcomes were death, clinically significant change in global function, mental state, relapse, hospital admission, adverse effects and acceptability of treatment. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data Peto odds ratios (OR) with the 95% confidence intervals (CI) were estimated. The number needed to treat statistic (NNT) was to have been calculated. Analysis was by intention-to treat. MAIN RESULTS: Four controlled clinical trials were found (total n=117). Smeraldi 1990 (n=20) compared bromperidol decanoate to placebo and found that more people in the latter group left the study by six months duration (50% versus 20%, OR 0.3 CI 0.05-7). There were no clear differences between bromperidol decanoate and placebo for a list of side effects. Ratings of global impression, mental state and needing additional antipsychotic medication all tended to favour the control depots (fluphenazine decanoate and haloperidol decanoate) and people consistently left the bromperidol decanoate group more frequently than those allocated other depots (n=97, OR 2.6 CI 0.8-9). There was no clear pattern in the occurrence of adverse effects. REVIEWER'S CONCLUSIONS: Currently, extrapolating from minimal trial data suggests that bromperidol decanoate may be better than a placebo injection but less valuable than fluphenazine or haloperidol decanoate. If bromperidol decanoate is available to the clinician it may be a viable choice, especially when there are reasons not to use fluphenazine or haloperidol decanoate. Well-conducted and reported randomised trials are urgently needed to inform practice in Belgium, Germany, Italy and the Netherlands. PMID- 10796448 TI - Depot pipothiazine palmitate and undeclynate for schizophrenia. AB - BACKGROUND: Anti-psychotic drugs are usually given orally but compliance with medication given by this route may be difficult to quantify. The development of depot injections in the 1960s gave rise to extensive use of depots as a means of long-term maintenance treatment. Pipothiazine palmitate is a depot from the phenothiazine family of antipsychotic drugs. OBJECTIVES: To assess the effects of depot pipothiazine palmitate and undeclynate versus placebo, oral anti-psychotics and other depot antipsychotic preparations for people with schizophrenia in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Relevant trials were identified by searching Biological Abstracts (1982-1998), Cochrane Library (Issue 2, 1998), Cochrane Schizophrenia Group's Register (June 1998), EMBASE (1980 1998), MEDLINE (1966-1998) and PsycLIT (1974-1998). References of all identified trials were also inspected for more studies and industry contacted. SELECTION CRITERIA: All clinical randomised trials focusing on people with schizophrenia where depot pipothiazine palmitate and undeclynate, oral anti-psychotics or other depot preparations were compared. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data Peto odds ratios (OR) with the 95% confidence intervals (CI) were estimated. Where possible, the number needed to treat statistic (NNT) was calculated. Analysis was by intention-to-treat. Normal continuous data were summated using the weighted mean difference (WMD). Scale data were presented only for those tools that had attained pre-specified levels of quality. MAIN RESULTS: Fourteen studies were included. When pipothiazine palmitate was compared to 'standard' oral antipsychotics no differences were found for outcomes of mental state, study attrition, behaviour and adverse effects (total randomised = 166). Pipothiazine palmitate was compared to other depot preparations in nine studies (n=455). Again no differences were identified for outcomes of global improvement, mental state, study attrition, behaviour and adverse effects. REVIEWER'S CONCLUSIONS: Although well-conducted and reported randomised trials are still needed to inform practice (no trial data exists reporting hospital and services outcomes, satisfaction with care and economics) pipothiazine palmitate is a viable choice for clinician and recipient of care. Data suggests it is not different to other depot antipsychotics. PMID- 10796449 TI - Injection therapy for subacute and chronic benign low back pain. AB - BACKGROUND: Injection with anaesthetics and/or steroids is one of the treatment modalities used in patients with chronic low back pain which needs evaluation with respect to the effectiveness on short and long term pain relief. OBJECTIVES: To evaluate the effectiveness of injection therapy in patients with low back pain lasting longer than one month. We distinguished between three injection sites: facet joint, epidural or local injections. SEARCH STRATEGY: We searched the Medline and Embase databases up to 1996 and other search methods as advocated by the Back Review Group search strategy. Abstracts and unpublished studies were not included. SELECTION CRITERIA: Randomized controlled trials of injection therapy for pain relief (although additional treatments were allowed) in patients with benign low back pain lasting longer than one month and not originating from cancer. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the trials for methodological quality. Subgroup analyses were made between trials with different control groups (placebo and active injections), with different injection site (facet joint, epidural and local injection), and timing of outcome measurement (short and long term). Within the resulting 12 subcategories of studies (2*3*2), the overall relative risks and corresponding 95% confidence intervals were estimated, using a random effects model (DerSimonian and Laird). In the case of trials in which control groups were active injections, we refrained from pooling the results. MAIN RESULTS: Twenty-one randomized trials were included in this review. All studies involved patients with low back pain lasting longer than one month. Only 11 studies compared injection therapy with placebo injections (explanatory trials). The methodologic quality of many studies was low: only 8 studies had a methodologic score of 50 or more points. There were only three well designed explanatory clinical trials: one on injections into the facet joints with a short-term RR of 0.89 (95% CI: 0.65-1.21) and a long-term RR of 0.90 (95% CI: 0.69-1.17); one on epidural injections with a short-term RR of 0.94 (95% CI: 0.76-1.15) and a long-term RR of 1.00 (95% CI: 0.71-1.41); and one on local injections with a long-term RR of 0.79 (95% CI: 0.65-0.96). Within the 6 subcategories of explanatory studies the pooled RRs with 95% confidence intervals were: facet joint, short-term: RR=0.89 (0.65-1.21); facet joint, long-term: RR=0.90 (0.69-1.17); epidural, short-term: RR=0.93 (0. 79-1.09); epidural, long term: RR=0.92 (0.76-1.11); local, short-term: RR=0.80 (0.40-1.59); local, long term: RR=0.79 (0.65-0. 96). REVIEWER'S CONCLUSIONS: Convincing evidence is lacking on the effects of injection therapies for low back pain. There is a need for more, well designed explanatory trials in this field. PMID- 10796450 TI - Nutritional supplementation for hip fracture aftercare in the elderly. AB - BACKGROUND: Fractures of the hip are an important cause of later ill health and mortality in elderly people. People with hip fractures are often malnourished at the time of fracture, and have poor food intake in hospital. OBJECTIVES: This review assesses the effects of nutritional interventions in elderly people recovering from hip fracture. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, the Cochrane Controlled Trials Register, Medline, Nutrition Abstracts and Reviews, Embase, Biosis, Cinahl, Healthstar and reference lists. We contacted investigators, and hand searched Proceedings of the Nutrition Society, Clinical Nutrition and The Journal of Parenteral and Enteral Nutrition. Date of the most recent search: June 1999. SELECTION CRITERIA: Randomised and quasi-randomised trials of nutritional interventions of mainly older patients (aged over 65 years) with hip fracture. DATA COLLECTION AND ANALYSIS: Trial allocation to included, excluded and awaiting assessment categories, was by consensus. Both reviewers independently extracted data and assessed trial quality. Additional information was sought from all trialists. Pooling of data for primary outcomes and select exploratory analyses were undertaken. MAIN RESULTS: Fifteen randomised trials involving 943 participants were included. Overall the quality of trials was poor; specifically in terms of allocation concealment, assessor blinding and intention to treat analysis. This, and the limited availability of outcome data, mean that the following results must be interpreted with caution. Oral multinutrient feeds (providing non-protein energy, protein, some vitamins and minerals), evaluated by six trials, may reduce unfavourable outcome (death or complications) (14/66 versus 26/73; Peto odds ratio 0.28, 95% confidence interval 0.11 to 0.67), but did not demonstrate an effect on mortality (12/91 versus 14/97; Peto odds ratio 0.82, 95% confidence interval 0.34 to 1.96). Four trials, examining nasogastric multinutrient feeding, showed no evidence for an effect on mortality (Peto odds ratio 0.98, 95% confidence interval 0.44 to 2.17), but the studies were heterogeneous regarding case-mix. Insufficient information was provided to evaluate unfavourable outcome. The effect of protein in an oral feed, tested in two trials, showed no evidence for an effect on mortality (Peto odds ratio 0.93, 95% confidence interval 0.31 to 2.79), but may have reduced days spent in rehabilitation wards. Two trials, testing intravenous thiamin (vitamin B1) and other water soluble vitamins, or 1-alpha-hydroxycholecalciferol (an active form of vitamin D) respectively, produced no evidence of benefit for either vitamin supplement. REVIEWER'S CONCLUSIONS: The strongest evidence for the effectiveness of nutritional supplementation exists for oral protein and energy feeds, but the evidence is still very weak. Future trials are required which overcome the defects of the reviewed studies, particularly inadequate size, methodology and outcome assessment. PMID- 10796451 TI - Carotid endarterectomy for asymptomatic carotid stenosis. AB - BACKGROUND: Whilst carotid endarterectomy (CEA) is of proven benefit in recently symptomatic patients with severe carotid stenosis, the role of carotid endarterectomy in preventing stroke in patients with asymptomatic carotid stenosis remains uncertain. OBJECTIVES: The objective of this review therefore was to determine the effects of CEA for patients with asymptomatic carotid stenosis. SEARCH STRATEGY: We searched the Cochrane Stroke Group Trials Register (June 1998), Medline (1966-Mar 1998), Current Contents (1995-Jan 1997), and reference lists of relevant articles. We contacted researchers in the field to identify additional published and unpublished studies. SELECTION CRITERIA: All completed randomised trials comparing CEA to medical treatment in patients with asymptomatic carotid stenosis. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data and assessed trial quality. Attempts were made to contact investigators to obtain missing information. MAIN RESULTS: Six trials were identified, but two were excluded on methodological grounds. Four trials with 2203 patients were included. In two trials aspirin was only given to patients in the medical group, and in two all patients received aspirin. The net excess "perioperative stroke or death" rate in the surgical group was 2.7% with relative risk 6.52 (95% confidence interval 2.66-15.96). The rates of "perioperative stroke or death or subsequent ipsilateral stroke" were 6.8% in the medical group vs 4.9% in the surgical group with RR 0.73 (0.52-1.02) favouring surgery. The rates of "any stroke or perioperative death" were 10.4% (medical) vs 8.1% (surgical) with RR 0.79 (0.60-1.02). The rates of "any stroke or death" were 23.2% (medical) vs 20.2% (surgical) with RR 0.89 (0.76-1.04). There were too few patients in CEA vs aspirin trials to determine whether aspirin had any confounding effect on outcome. An additional analysis including data from a fifth small unpublished trial altered slightly the risk ratios in favour of surgery and narrowed confidence intervals sufficiently to achieve statistical significance for each outcome. However, inclusion of these data had no appreciable effect on relative or absolute risk reduction. REVIEWER'S CONCLUSIONS: There is some evidence favouring CEA for asymptomatic carotid stenosis, but the effect is at best barely significant, and extremely small in terms of absolute risk reduction. PMID- 10796452 TI - Antiplatelet therapy for preventing stroke in patients with non-valvular atrial fibrillation and no previous history of stroke or transient ischemic attacks. AB - BACKGROUND: Atrial fibrillation (AF) carries an increased risk of stroke; antiplatelet agents are proven effective for stroke prevention in other settings. OBJECTIVES: The objective of this review was to determine the efficacy and safety of antiplatelet therapy for prevention of stroke in patients with chronic non valvular AF. SEARCH STRATEGY: We searched the Cochrane Stroke Group Specialised Register of Trials, MEDLINE database (June 1999), and the database of the Antithrombotic Trialists Collaboration, as well as reference lists of relevant articles. SELECTION CRITERIA: All randomized trials comparing antiplatelet therapies to placebo in patients with non-valvular AF and no history of transient ischemic attack (TIA) or stroke. DATA COLLECTION AND ANALYSIS: Trials for inclusion were independently selected by two reviewers who also extracted each outcome and double-checked the data. The Peto method was used for combining odds ratios. All analysis were, as far as possible, "intention-to-treat". Since the published results of two trials included 3-8% of participants with prior stroke or TIA, unpublished results excluding these participants were obtained from the Atrial Fibrillation Investigators. MAIN RESULTS: Among 1680 participants without prior stroke/TIA, randomized to aspirin (N = 838) or placebo in two trials, aspirin was associated with nonsignificantly lower risks of ischemic stroke (OR = 0.71, CI 95% 0. 46 - 1.10), all stroke (OR = 0.70, CI 95% 0.45 - 1.08) all disabling/fatal stroke (OR =0.88, CI 95% 0.48 - 1.58) and the constellation of stroke, MI or vascular death (OR = 0.76, CI 95% 0. 54 - 1.05 ). Considering all randomized participants including those with prior stroke or TIA, reductions in these events by aspirin were consistently smaller and marginally statistically significant: ischemic stroke (OR = 0.77, CI 95% 0.60-1.00), all stroke (OR = 0.76, CI 95% 0.61 - 0.93), all disabling/fatal stroke (OR = 0.87, CI 95% 0.64 - 1.19) and the combined outcome (OR = 0.79, CI 95% 0.64 - 0. 99). No increase in major hemorrhage was seen, but the number of hemorrhagic events was small. REVIEWER'S CONCLUSIONS: Considering all randomized data, aspirin modestly (by about 20%) reduces stroke and major vascular events in nonvalvular AF. For primary prevention among AF patients with an average stroke rate of 4.5%/year, about 10 strokes would be prevented yearly for every 1000 given aspirin. PMID- 10796453 TI - Oral anticoagulants for preventing stroke in patients with non-valvular atrial fibrillation and no previous history of stroke or transient ischemic attacks. AB - BACKGROUND: Non-valvular atrial fibrillation (AF) is associated with an increased risk of stroke. OBJECTIVES: The objective of this review was to characterize the efficacy and safety of oral anticoagulation (OAC) with vitamin K antagonists for the primary prevention of stroke in patients with chronic AF. SEARCH STRATEGY: We searched the Cochrane Stroke Group Specialised Register of Trials (June 1999), MEDLINE database, and the database of the Antithrombotic Trialists Collaboration, as well as reference lists of relevant articles. SELECTION CRITERIA: All randomized controlled trials comparing the value of OAC versus control in patients with non-valvular chronic atrial fibrillation and no history of transient ischemic attack (TIA) or stroke. DATA COLLECTION AND ANALYSIS: Trials for inclusion were independently selected by two reviewers who also extracted each outcome and double-checked the data. The Peto method was used for combining odds ratios. All analysis were, as far as possible, "intention-to-treat". Since the published results of four trials included 3-8% of participants with prior stroke or TIA, unpublished results excluding these participants were obtained from the Atrial Fibrillation Investigators. MAIN RESULTS: Of 2313 participants without prior cerebral ischemia from five trials, about half (n = 1154) were randomized to adjusted-dose OAC with an estimated mean INRs ranging between 2.0 2.6 during 1.5 years/participant average follow-up. Participant features and study quality were similar between trials. OAC was associated with large, highly statistically significant reductions in ischemic stroke (OR = 0.34, 95% CI 0.23 - 0.52), all stroke (OR = 0.39, 95% CI 0.26 - 0. 59), all disabling or fatal stroke (OR = 0.47, 95% CI 0.28 - 0.80), and the combined endpoint of all stroke, MI or vascular death (OR = 0.56, 95% CI 0.42 - 0.76). The observed rates of intracranial and extracranial hemorrhage not significantly increased by OAC therapy, but confidence intervals were wide. REVIEWER'S CONCLUSIONS: Adjusted dose OAC (achieved INRs between 2-3) reduces stroke as well as disabling/fatal stroke for patients with nonvalvular AF, and these benefits were not substantially offset by increased bleeding among participants in randomized clinical trials. Limitations include relatively short follow-up and imprecise estimates of bleeding risks from these selected participants. For primary prevention in AF patients who have an average stroke rate of 4%/year, about 25 strokes and about 12 disabling fatal strokes would be prevented yearly for every 1000 given OAC. PMID- 10796454 TI - Calcium antagonists for acute ischemic stroke. AB - BACKGROUND: The sudden loss of blood supply in ischemic stroke is associated with increased levels of calcium ions within neurones. Inhibiting this increase could protect neurones and is thought to reduce neurological impairment, disability and handicap after stroke. OBJECTIVES: The aim of this review is to determine whether calcium antagonists reduce the risk of death or dependency after acute ischemic stroke. The influence of different drugs, dosages, routes of administration, time intervals after stroke and trial design on the risk of poor outcome was investigated. SEARCH STRATEGY: Relevant trials were identified in the Specialised Register of Controlled Trials (last searched: March 1999). SELECTION CRITERIA: All truly randomised trials comparing a calcium antagonist with control in patients with acute ischaemic stroke were included. DATA COLLECTION AND ANALYSIS: Two authors assessed all trials and extracted the data. Poor outcome, defined as death or dependency in activities of daily living, was used as the main outcome. Analyses were, if possible, "intention-to-treat". MAIN RESULTS: 46 trials were identified of which 28 were included (7521 patients). No effect of calcium antagonists on poor outcome at the end of follow-up (OR 1.07, 95% CI 0.97/1.18), or on death at end of follow-up (OR 1.10, 95% CI 0.98/1.24) was found. Intravenous administration (i.v.) of calcium antagonists was associated with an increase in the number of patients with poor outcome compared to oral administration (indirect comparisons). Comparisons of different doses of nimodipine suggested that the highest doses were associated with poorer outcome. Administration within 12 hours of onset was associated with an increase in the proportion of patients with poor outcome, but this effect was largely due to the poor results associated with i.v. administration. A subgroup analysis on nimodipine (oral, 120 mg/day) started within 12 hours of stroke onset, did not show a beneficial effect. REVIEWER'S CONCLUSIONS: No evidence is available to justify the use of calcium antagonists in patients with acute ischaemic stroke. PMID- 10796455 TI - Loxapine for schizophrenia. AB - BACKGROUND: Loxapine is a drug with D2/D3 receptor antagonist activity and a higher affinity for D3 than D2. Some authors have suggested that loxapine is more effective than typical antipsychotics in reducing the negative symptoms of schizophrenia, that extrapyramidal side-effects are not usually seen at clinically effective antipsychotic doses and that it should therefore be classed as atypical. OBJECTIVES: To determine the effects of loxapine compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), EMBASE (1980-1999), MEDLINE (1966-1999), LILACS (1982-1996), PSYNDEX (1977-1995) and PsycLIT (1974-1999) were undertaken. In addition, pharmaceutical databases on the Dialog Corporation Datastar and Dialog services were searched. References of all identified studies were searched for further trials. Pharmaceutical companies and authors of trials were contacted. SELECTION CRITERIA: All randomised controlled clinical trials relevant to the care of schizophrenia that compared loxapine to other treatments. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted but excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the risk ratio (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data, weighted mean differences were calculated (WMD). All data were inspected for heterogeneity. MAIN RESULTS: Compared to placebo, loxapine is antipsychotic (Global effect - not improved at 6 weeks, n=66, RR 0.6 CI 0.4-0.9 NNT 4 CI 2-62) with similar adverse effect profile to typical drugs. Is as effective as typical drugs in the short term (4-12 weeks) (Global effect - not improved, n=411, RR 0.89 CI 0.7-1.2). Very limited heterogeneous data suggest that, given intramuscularly (IM), loxapine may be at least as sedating as IM haloperidol and thiothixene. REVIEWER'S CONCLUSIONS: Loxapine is antipsychotic but its effects are under researched. It is not clearly different from typical drugs in either its positive or adverse effects. PMID- 10796456 TI - Vancomycin for prophylaxis against sepsis in preterm neonates. AB - BACKGROUND: Nosocomial, late onset sepsis occurs in up to 50% of infants of less than 1000gm at birth. The commonest organism isolated is coagulase negative staphylococcus (CoNS). A number of studies have evaluated the efficacy or prophylactic low dose vancomycin given either as a continuous infusion added to the infant's hyperalimentation fluid or by intermittent intravenous administration and these studies in very low birth weight infants are the subject of this review. OBJECTIVES: To evaluate the safety and efficacy of vancomycin prophylaxis for the prevention of late-onset sepsis, coagulase negative staphylococcal sepsis, mortality, and effects on length of stay, total vancomycin exposure, evidence of vancomycin toxicity, and the development of vancomycin resistant organisms in the preterm neonate. SEARCH STRATEGY: Searches were made of Medline, (MeSH terms: Vancomycin and Sepsis; limits: age groups, newborn infants), HealthStar and EMBase, electronic abstracts, personal files and conference proceedings. SELECTION CRITERIA: Randomized controlled trials which compared the incidence of sepsis and mortality in preterm neonates receiving vancomycin prophylaxis versus a control group receiving no prophylaxis. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including the overall incidence of sepsis, the incidence of coagulase negative staphylococcal sepsis, mortality, length of stay, total vancomycin exposure, evidence of vancomycin toxicity, and the development of vancomycin resistant organisms were excerpted from previous clinical trials. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: The administration of prophylactic vancomycin reduced the incidence of both total neonatal nosocomial sepsis and coagulase negative staphylococcal sepsis in eligible preterm infants. Mortality, length of stay, and evidence of vancomycin toxicity were not significantly different between the two groups. There was insufficient evidence to ascertain the risks of development of vancomycin resistant organisms in the nurseries involved in these trials. REVIEWER'S CONCLUSIONS: The use of prophylactic vancomycin in low doses reduces the incidence of nosocomial sepsis in the neonate. The methodologies of these studies may have contributed to the low rate of sepsis in the treated groups, as the blood cultures drawn from central lines may have failed to grow due to the low levels of vancomycin in the infusate. Although there is a theoretical concern regarding the development of resistant organisms with the administration of prophylactic antibiotic, there is insufficient evidence to ascertain the risks of development of vancomycin resistant organisms. Few clinically important benefits have been demonstrated for very low birth weight infants treated with prophylactic vancomycin. It therefore appears that routine prophylaxis with vancomycin should not be undertaken at present. PMID- 10796457 TI - Calcitonin for the treatment and prevention of corticosteroid-induced osteoporosis. AB - BACKGROUND: Corticosteroid-induced osteoporosis is a cause of morbidity in patients with chronic obstructive lung disease, asthma, and rheumatologic disorders. Corticosteroid treatment causes bone loss by a variety of complex mechanisms. It has been shown that bone mineral loss at the hip averages 14% in the first year after starting corticosteroid therapy. OBJECTIVES: To review the efficacy of calcitonin (subcutaneous or nasal) for the treatment and prevention of corticosteroid-induced osteoporosis. SEARCH STRATEGY: We conducted a search of Medline, the Cochrane Controlled Trials Register and Embase using the Cochrane Musculoskeletal Group search strategy for randomized controlled trials (RCTs) up to May 1998. We also searched bibliographic references and consulted content experts. SELECTION CRITERIA: Two independent reviewers selected RCTs which met predetermined inclusion criteria. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data using predetermined forms and assessed methodological quality of randomization, blinding and dropouts. For dichotomous outcomes, relative risks (RR) were calculated. For continuous data, weighted mean differences (WMD) of the percent change from baseline were calculated. We decided a priori to use random effects models for all outcomes, because of uncertainty about whether a consistent true effect exists in such different populations. MAIN RESULTS: Nine trials met the inclusion criteria, including 221 patients randomized to calcitonin and 220 to placebo. The median methodologic quality was two out of a maximum of five points. Calcitonin was more effective than placebo at preserving bone mass at the lumbar spine after six and 12 months of therapy with a WMD of 2.8% (95% CI: 1.4 to 4.3) and 3.2% (95% CI: 0.3 to 6.1). At 24 months, lumbar spine BMD was not statistically different between groups: WMD 4.5% (95% CI: -0.6 to 9.5)]. Bone density at the distal radius was also higher with calcitonin after six months of therapy, but bone density at the femoral neck was not different between placebo and calcitonin treated groups. The relative risk of fractures was not significantly different between calcitonin and placebo with a relative risk (RR) of 0.71 (95% CI: 0.26 to 1.89) for vertebral and 0.52 (95% CI: 0.14 to 1.96) for nonvertebral fractures. The subgroup analyses of methodological quality and duration of corticosteroid therapy were confounded. Trials of patients who had been taking steroids for greater than three months (which were of low methodologic quality) demonstrated a larger effect of calcitonin on spine bone density (about 6%) than prevention trials (about 1%). There was no consistent effect of different dosages (50-100 IU compared to 200-400 IU). However, subcutaneous calcitonin showed substantially greater prevention of bone loss. Withdrawals due to side effects were higher in the calcitonin-treated groups: RR 3.19 (95%CI: 0.66 to 15.47). Important side effects included nausea and facial flushing. REVIEWER'S CONCLUSIONS: Calcitonin appears to preserve bone mass in the first year of glucocorticoid therapy at the lumbar spine by about 3% compared to placebo, but not at the femoral neck. Our analysis suggests that the protective effect on bone mass may be greater for the treatment of patients who have been taking corticosteroids for more than three months. Efficacy of calcitonin for fracture prevention in steroid-induced osteoporosis remains to be established. PMID- 10796458 TI - Multidisciplinary rehabilitation for fibromyalgia and musculoskeletal pain in working age adults. AB - BACKGROUND: Non-malignant musculoskeletal pain is an increasing problem in western countries. Fibromyalgia syndrome is an increasing recognised chronic musculoskeletal disorder. OBJECTIVES: The objective of this systematic review was to determine the effectiveness of multidisciplinary rehabilitation for fibromyalgia and widespread musculoskeletal pain among working age adults. SEARCH STRATEGY: An electronic search was conducted and included Medline from 1966, PsycLIT from 1967 and EMBASE from 1980 to April 1998. The Cochrane Musculoskeletal Group Trials Register was searched as well as, the Cochrane Controlled Trials Register (CCTR). The references of identified articles and reviews were checked, studies published in the Finnish medical database Medic from 1978 to 1998 screened and the Science Citation Index searched. Content experts were also contacted for additional or unpublished studies. SELECTION CRITERIA: From all references found in our original search, we selected all randomized controlled trials (RCTs) and clinical controlled trials (CCTs). Trials had to assess the effectiveness of multidisciplinary rehabilitation for patients suffering from fibromyalgia and widespread musculoskeletal pain among working age adults. The rehabilitation program was required to be multidisciplinary; that is, it had to consist of a physician's consultation, plus a psychological, social or vocational intervention, or a combination of both. DATA COLLECTION AND ANALYSIS: Four reviewers independently selected the RCTs and CCTs that met the specified inclusion criteria. Two experts in the field of rehabilitation evaluated the relevance and applicability of the findings of the selected studies to actual clinical use. Two other reviewers extracted the data and assessed the main results and the methodological quality of the studies using standardized forms. Finally, a qualitative analysis was performed to evaluate the level of scientific evidence for the effectiveness of multidisciplinary rehabilitation. MAIN RESULTS: After screening 1808 abstracts, and the references of 65 reviews, we found only seven relevant studies (1050 patients) that met our inclusion criteria. None of these were considered, methodologically, a high quality randomized controlled trial. Four of the included RCTs on fibromyalgia were graded low quality and suggest no quantifiable benefits. The three included RCTs on widespread musculoskeletal pain showed that based on limited evidence, overall, no evidence of efficacy was observed. However, behavioral treatment and stress management appear to be important components. Education combined with physical training showed some positive effects in long term follow up. REVIEWER'S CONCLUSIONS: We conclude that there appears to be little scientific evidence for the effectiveness of multidisciplinary rehabilitation for these musculoskeletal disorders. However, multidisciplinary rehabilitation is a commonly used intervention for chronic musculoskeletal disorders, which cause much personal suffering and substantial economic loss to the society. There is a need for high quality trials in this field. PMID- 10796459 TI - Behavioural treatment for chronic low back pain. AB - BACKGROUND: The treatment of chronic low back pain is not primarily focused on removing an underlying organic pathology, but at the reduction of disability through the modification of environmental contingencies and cognitive processes. Behavioural interventions are commonly used in the treatment of chronic (disabling) low back pain. OBJECTIVES: The objective of this systematic review was to determine if behavioural therapy is more effective than reference treatments for chronic non-specific low back pain, and which type of behavioural treatment is most effective. SEARCH STRATEGY: We searched the Medline, PsycLit databases, and the Cochrane Controlled Trials Register up to April 1999, Embase up to September 1999. We also screened references of identified randomised trials and relevant systematic reviews. SELECTION CRITERIA: Only randomised trials on any type of behavioural treatment for non-specific chronic low back pain were included. DATA COLLECTION AND ANALYSIS: Methodological quality assessment and data extraction was done by two reviewers independently. The magnitude of effect was assessed by computing a pooled effect size for each domain (i.e., behavioural outcomes, overall improvement, back pain specific and generic functional status, return to work, and pain intensity) using the random effects model. MAIN RESULTS: Only 6 studies (25%) were high quality. There is strong evidence (level 1) that behavioural treatment has a moderate positive effect on pain intensity (pooled effect size 0.62; 95% CI 0.25, 0.98), and small positive effects on generic functional status (pooled effect size 0.35; 95% CI -0.04, 0.74) and behavioural outcomes (pooled effect size 0.40; 95% CI 0.10, 0.70) of chronic low back pain patients when compared to waiting list controls or no treatment. There is moderate evidence (level 2) that an additional behavioural component to a usual treatment program for chronic low back pain has no positive short-term effect on generic functional status (pooled effect size 0.31; 95% CI - 0.01, 0.64), pain intensity (pooled effect size 0.03; 95% CI - 0.30, 0. 36) and behavioural outcomes (pooled effect size 0.19; 95% CI - 0.08, 0.45). REVIEWER'S CONCLUSIONS: Behavioural treatment seems to be an effective treatment for chronic low back pain patients, but it is still unknown what type of patients benefit most from what type of behavioural treatment. PMID- 10796460 TI - Low level laser therapy (classes I, II and III) for the treatment of osteoarthritis. AB - BACKGROUND: Osteoarthritis (OA) affects a large proportion of the population. Low Level Laser Therapy (LLLT) is a light source that generates extremely pure light, of a single wavelength. The effect is not thermal, but rather related to photochemical reactions in the cells. LLLT was introduced as an alternative non invasive treatment for OA about 10 years ago, but its effectiveness is still controversial. OBJECTIVES: To assess the effectiveness of LLLT in the treatment of OA. SEARCH STRATEGY: We searched MEDLINE, EMBASE, the Cochrane Musculoskeletal registry, the registry of the Rehabilitation and Related Thereapies field and the Cochrane Controlled Trials Register up to January 30, 2000. SELECTION CRITERIA: Following an a priori protocol, only controlled clinical trials of LLLT for the treatment of patients with a clinical diagnosis of OA were eligible. Abstracts were excluded unless further data could be obtained from the authors. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials and abstracted data using predetermined forms. Heterogeneity was tested with Cochran's Q test. A fixed effects model was used throughout for continuous variables, except where heterogeneity existed, in which case, a random effects model was used. Results were analyzed as weighted mean differences (WMD) with 95% confidence intervals (CI), where the difference between the treated and control groups was weighted by the inverse of the variance. Standardized mean differences (SMD) were calculated by dividing the difference between treated and control by the baseline variance. SMD were used when different scales were used to measure the same concept (e.g. pain). Dichotomous outcomes were analyzed with odds ratios. MAIN RESULTS: Five trials were included, with 112 patients randomized to laser, 85 patients to placebo laser. Treatment duration ranged from 4 to 10 weeks. Pain was assessed by four trials. The pooled estimate (random effects) of three trials showed no effect on pain measured using a scale (SMD: -0.2, 95% CI: 1.0, +0.6), but there was statistically significant heterogeneity (p>0,05). Two of the trials showed no effect and one demonstrated very beneficial effects with laser. In another trial, with no scale-based pain outcome, significantly more patients reported pain relief (yes/no) with laser with an odds ratio of 0.05, (95% CI: 0.0 to 1.56). Other outcomes of joint tenderness, joint mobility and strength were not significant. REVIEWER'S CONCLUSIONS: For OA, the results are conflicting in different studies and may depend on the method of application and other features of the LLLT application. Clinicians and researchers should consistently report the characteristics of the LLLT device and the application techniques used. New trials on LLLT should make use of standardized, validated outcomes. Despite some positive findings, this meta-analysis lacked data on how LLLT effectiveness is affected by four important factors: wavelength, treatment duration of LLLT, dosage and site of application over nerves instead of joints. There is clearly a need to investigate the effects of these factors on LLLT effectiveness for OA in randomized controlled clinical trials. PMID- 10796461 TI - Auranofin versus placebo in rheumatoid arthritis. AB - BACKGROUND: Auranofin is an oral gold compound used for the treatment of rheumatoid arthritis RA. The use of auranofin has declined in the past few years, perhaps due in part to conflicting results from different studies. OBJECTIVES: To estimate the short-term efficacy and toxicity of auranofin for the treatment of (RA) SEARCH STRATEGY: We searched MEDLINE and EMBASE up to December 1998, the Cochrane Controlled Trials Register (CCTR) version 4, 1998, and the Cochrane Musculoskeletal Group Specialized Register. A hand search was also done of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing auranofin against placebo in patients with RA DATA COLLECTION AND ANALYSIS: The methodological quality of the trials was assessed using a validated assessment tool (Jadad 1996). Rheumatoid arthritis outcome measures were extracted from the publications for the 6-month endpoint. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts, pain and global assessments. The weighted mean difference (WMD) was used for ESR. Toxicity was evaluated with pooled odds ratios for withdrawals and adverse reactions. A chi-square test was used to assess heterogeneity among trials. In the presence of heterogeneity, random effects models were used. Otherwise, the data was pooled assuming fixed effects. MAIN RESULTS: A statistically significant benefit was observed for auranofin when compared to placebo for tender joint scores, pain, patient and physician global assessments and ESR. The SMD between treatment and placebo was -0.39 (95% CI -0.54, -0.25) for tender joint scores, -0.08 (95% CI -0.22, -0.07) for swollen joint scores, and the WMD was -4.68 (95% CI -6.59, -2.77) for pain scores and -9.85mm (95% CI 16.46, -3.25) for ESR. Withdrawals from adverse reactions were 1.5 times higher in the auranofin group OR = 1.52 (95% CI 0.94, 2.46) but this result was not statistically significant. Patients receiving placebo were three times more likely to discontinue treatment because of lack of efficacy than patients receiving auranofin: OR=0.31 (95% CI: 0.21, 0.44). REVIEWER'S CONCLUSIONS: Auranofin appears to have a small clinically and statistically significant benefit on the disease activity of patients with RA. The beneficial effects appear to be modest compared to drugs such as methotrexate or parenteral gold. Its effects on long term health status and radiological progression are not clear at this time. PMID- 10796462 TI - Low level laser therapy (classes I, II and III) in the treatment of rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) affects a large proportion of the population. Low Level Laser Therapy (LLLT) was introduced as an alternative non invasive treatment for RA about 10 years ago. LLLT is a light source that generates extremely pure light, of a single wavelength. The effect is not thermal, but rather related to photochemical reactions in the cells. The effectiveness of LLLT for rheumatoid arthritis is still controversial. OBJECTIVES: To assess the effectiveness of LLLT in the treatment of RA. SEARCH STRATEGY: We searched MEDLINE, EMBASE, the registries of the Cochrane Musculoskeletal group and the field of Rehabilitation and Related Therapies as well as the Cochrane Controlled Trials Register up to January 30, 2000. SELECTION CRITERIA: Following an a priori protocol, we selected only randomized controlled trials of LLLT for the treatment of patients with a clinical diagnosis of RA were eligible. Abstracts were excluded unless further data could be obtained from the authors. DATA COLLECTION AND ANALYSIS: Two reviewers independently select trials for inclusion, then extracted data and assessed quality using predetermined forms. Heterogeneity was tested with Cochran's Q test. A fixed effects model was used throughout for continuous variables, except where heterogeneity existed, in which case, a random effects model was used. Results were analyzed as weighted mean differences (WMD) with 95% confidence intervals (CI), where the difference between the treated and control groups was weighted by the inverse of the variance. Standardized mean differences (SMD) were calculated by dividing the difference between treated and control by the baseline variance. SMD were used when different scales were used to measure the same concept (e.g. pain). Dichotomous outcomes were analyzed with odds ratios. MAIN RESULTS: A total of 204 patients were included in the five placebo-controlled trials, with 112 randomized to laser therapy. Relative to a separate control group, LLLT reduced pain by 70% relative to placebo and reduced morning stiffness duration by 27.5 minutes (95%CI: 2.9 to 52 minutes) and increased tip to palm flexibility by 1.3 cm (95% CI: 0. 8 to 1.7 cm). Other outcomes such as functional assessment, range of motion and local swelling did not differ between groups. There were no significant differences between subgroups based on LLLT dosage, wavelength, site of application or treatment length. For RA, relative to a control group using the opposite hand, there was no difference between the control and treatment hand, but all hands improved in terms of pain relief and disease activity. REVIEWER'S CONCLUSIONS: In summary, LLLT for RA is beneficial as a minimum of a four-week treatment with reductions in pain and morning stiffness. On the one hand, this meta-analysis found that pooled data gave some evidence of a clinical effect, but the outcomes were in conflict, and it must therefore be concluded that firm documentation of the application of LLLT in RA is not possible. Clinicians and researchers should consistently report the characteristics of the LLLT device and the application techniques used. New trials on LLLT should make use of standardized, validated outcomes. Despite some positive findings, this meta analysis lacked data on how LLLT effectiveness is affected by four important factors: wavelength, treatment duration of LLLT, dosage and site of application over nerves instead of joints. PMID- 10796463 TI - Surfactant for meconium aspiration syndrome in full term infants. AB - OBJECTIVES: To evaluate the effect of surfactant administration in the treatment of term infants with meconium aspiration syndrome. SEARCH STRATEGY: Searches were made using Medline (1985 to January 2000) (MeSH terms: pulmonary surfactant and meconium aspiration; limits: age groups, newborns; publication type, clinical trials), previous reviews including cross-references, abstracts, conference and symposia proceedings, expert informants, and journal hand searching in the English language. Authors were directly contacted to provide additional data. SELECTION CRITERIA: Randomized controlled trials which evaluated the effect of surfactant administration in term infants with meconium aspiration syndrome are included in the analysis. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including duration of assisted ventilation, duration of supplemental oxygen, pneumothorax, intraventricular hemorrhage (any grade and severe IVH), chronic lung disease, treatment with extracorporeal membrane oxygenation (ECMO), and mortality were excerpted from the reports of the clinical trails by the reviewers. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Two randomized controlled trials met inclusion criteria. Findlay (1996) reports a decrease in the risk of pneumothorax (relative risk 0.09, 95% CI 0.01, 1.54, risk difference -0.25, 95% CI -0.45, -0.05). Both Findlay (1996) and Lotze (1998) report a decrease in the number of infants receiving extracorporeal membrane oxygenation. The meta analysis supports a significant reduction in the risk of requiring extracorporeal membrane oxygenation (typical relative risk 0.64, 95% CI 0.46, 0.91 typical risk difference -0.17, 95% CI -0.30, -0.04). No difference was noted in overall mortality (typical relative risk 1.86 95% CI 0.35, 9.89, typical risk difference 0.02 95% CI -0.03, 0.07). REVIEWER'S CONCLUSIONS: In infants with meconium aspiration syndrome, surfactant administration may reduce the severity of respiratory illness and decrease the number of infants with progressive respiratory failure requiring support with ECMO. The relative efficacy of surfactant therapy compared to, or in conjunction with, other approaches to treatment including inhaled nitric oxide, liquid ventilation, and high frequency ventilation remains to be tested. PMID- 10796464 TI - Molindone for schizophrenia and severe mental illness. AB - BACKGROUND: Typical antipsychotic drugs are widely used as the first line treatment for people with schizophrenia. However, the atypical class of antipsychotic drugs is making important inroads into this approach. 'Atypical' is a term widely used to describe some antipsychotics which have a low propensity to produce movement disorders, sedation and raised serum prolactin. There is some suggestion that the different adverse effect profiles of the atypical antipsychotic group make them more acceptable to people with schizophrenia. Molindone has a similar profile to quetiapine (a novel atypical antipsychotic), with very low binding to all receptors. Some authors have suggested that molindone is safer than other 'typical' antipsychotics in that extrapyramidal adverse effects are not usually seen at clinically effective antipsychotic doses and that it should therefore be classed as an atypical antipsychotic. OBJECTIVES: To determine the effects of molindone compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), The Cochrane Library CENTRAL (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), CINAHL (1982-1999), EMBASE (1980-1999), MEDLINE (1966-1999), LILACS (1982-1999), PSYNDEX (1977-1999), and PsycLIT (1974-1999) were undertaken. In addition, pharmaceutical databases on the Dialog Corporation Datastar and Dialog services were searched. References of all identified studies were searched for further trials. The manufacturer of molindone and authors of trials were contacted. SELECTION CRITERIA: All randomised controlled trials that compared molindone to other treatments for schizophrenia and schizophrenia-like psychoses were included by independent assessment. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. Data were excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the risk ratio (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to treat basis. For continuous data, weighted mean differences (WMD) were calculated. All data were inspected for heterogeneity. MAIN RESULTS: Thirteen studies were included in the review. Data for this compound range from very short (10 day) studies of the intramuscular preparation to trials lasting over three months. For measures of global state available data do not justify any conclusions on the comparative efficacy of molindone and placebo. When compared to other typical antipsychotics no difference in effectiveness was evidenced (doctors' RR 1.13, CI 0.69 to 1.86; nurses' RR 1.23, CI 0.82 to 1.86). It is no more or less likely than typical drugs to cause movement disorders, but causes significantly more weight loss (RR 2.78, CI 1.10 to 6.99). REVIEWER'S CONCLUSIONS: The strength of the evidence relating to this compound is limited, owing to small sample size, poor study design, limited outcomes and incomplete reporting. Molindone may be an effective antipsychotic; however, its adverse effect profile does not differ significantly from that of typical antipsychotics, apart from the event of weight loss. At present there is no evidence to suggest that it may have an atypical profile. PMID- 10796465 TI - Acupuncture for chronic asthma. AB - BACKGROUND: Acupuncture has traditionally been used to treat asthma in China and is used increasingly for this purpose internationally. OBJECTIVES: The objective of this review was to assess the effects of acupuncture for the treatment of asthma or asthma-like symptoms. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, the Cochrane Complementary Medicine Field trials register and reference lists of articles. SELECTION CRITERIA: Randomised and possibly randomised trials using acupuncture to treat asthma and asthma-like symptoms. Acupuncture could involve the insertion of needles or other forms of stimulation of acupuncture points. DATA COLLECTION AND ANALYSIS: Trial quality was assessed by at least two reviewers independently. A reviewer experienced in acupuncture assessed the adequacy of the sham acupuncture. Study authors were contacted for missing information. MAIN RESULTS: Seven trials involving 174 people were included. Trial quality varied and results were inconsistent. No statistically significant or clinically relevant effects were found for acupuncture compared to sham acupuncture. However the points used in the sham arm of some studies are used for the treatment of asthma according to traditional Chinese medicine. Only one study used individualised treatment strategies. Lung function could be compared statistically in only 3 trials. Peak expiratory flow rate showed a statistically insignificant increase of 8.4 litres/minute weighted mean difference (95% confidence interval -29.4 to 46.2) when acupuncture was compared to sham acupuncture. REVIEWER'S CONCLUSIONS: There is not enough evidence to make recommendations about the value of acupuncture in asthma treatment. Further research needs to consider the complexities and different types of acupuncture. PMID- 10796466 TI - Acupuncture for smoking cessation. AB - BACKGROUND: Acupuncture is promoted as a treatment for smoking cessation, and is believed to reduce withdrawal symptoms. OBJECTIVES: The objective of this review is to determine the effectiveness of acupuncture in smoking cessation in comparison with: a) sham acupuncture b) other interventions c) no intervention. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register, Medline, PsycLit, Dissertation Abstracts, Health Planning and Administration, Social SciSearch, Smoking & Health, Embase, Biological Abstracts and DRUG. SELECTION CRITERIA: Randomised trials comparing a form of acupuncture with either sham acupuncture, another intervention or no intervention for smoking cessation. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the nature of the acupuncture and control procedures, the outcome measures, method of randomisation, and completeness of follow-up. We assessed abstinence from smoking at the earliest time-point (before 6 weeks), at six months and at one year follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Those lost to follow-up were counted as continuing to smoke. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: We identified 18 publications involving 20 comparisons. Acupuncture was not superior to sham acupuncture in smoking cessation at any time point. The odds ratio (OR) for early outcomes was 1.22 (95% confidence interval 0.99 to 1.49); the OR after 6 months was 1.38 (95% confidence interval 0.90 to 2.11) and after 12 months 1.02 (95% confidence interval 0.72 to 1.43). Similarly, when acupuncture was compared with other anti-smoking interventions, there were no differences in outcome at any time point. Acupuncture appeared to be superior to no intervention in the early results, but this difference was not sustained. The results with different acupuncture techniques do not show any one particular method (i.e. auricular acupuncture or non-auricular acupuncture) to be superior to control intervention. REVIEWER'S CONCLUSIONS: There is no clear evidence that acupuncture is effective for smoking cessation. PMID- 10796467 TI - Interventions for promoting adherence to tuberculosis management. AB - BACKGROUND: Up to half the people with tuberculosis do not complete their treatment. Strategies to improve adherence to treatment can be targeted at the person with the disease or at health workers. OBJECTIVES: The objective of this review was to assess the effects of promoting adherence to anti-tuberculosis treatment and completion of diagnostic protocols for TB. SEARCH STRATEGY: We searched the Cochrane Collaboration Trials, the Cochrane Effective Professional Practice Group trials register, the Cochrane Infectious Diseases Group trials register, Medline, Embase, Lilacs and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of interventions to promote adherence with curative or preventive chemotherapy and diagnostic protocols for tuberculosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Eleven trials were included. Reminder cards sent to defaulters, a combination package of a monetary incentive and health education and more supervision of tuberculosis clinic staff increased the number of people completing their tuberculosis treatment. Direct observation by clinic nurses of people swallowing their tuberculosis drugs did not increase the likelihood of treatment success. Return to the clinic for reading of a tuberculin skin test was enhanced by monetary incentives, assistance by lay health workers, contracts and telephone prompts but not by health education. REVIEWER'S CONCLUSIONS: We have found evidence of benefit for a number of specific interventions to improve adherence to anti-tuberculous therapy and completion of diagnostic protocols. These should be implemented by health care providers where appropriate to local circumstances. Future studies in low income countries are a priority and should measure adherence as well as clinical outcomes. PMID- 10796468 TI - Amodiaquine for treating malaria. AB - BACKGROUND: Amodiaquine has been widely used to treat malaria. Due to reports of fatal adverse drug reactions, discontinuation or modification of its use has been suggested. OBJECTIVES: The objective of this review was to assess the effects of amodiaquine for treating malaria. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register and Medline. We also contacted researchers in the field and drug companies. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing amodiaquine with other treatment for uncomplicated malarial infections in adults and children. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Forty trials were included. Allocation was adequately concealed in three trials. Amodiaquine was more effective than chloroquine for parasite clearance. The combined results of parasite clearance at seven days from 24 trials was 83% for amodiaquine and 56% for chloroquine (odds ratio 4.29, 95% confidence interval 3.51 to 5.24). The odds ratio for parasite clearance at 14 days was 6.00, 95% confidence interval 4.38 to 8.21. Amodiaquine and sulfadoxine/pyrimethamine showed similar results for parasite clearance on day seven, but sulfadoxine/pyrimethamine appeared to be more effective on day 14 and 28. No significant difference for adverse events was observed between amodiaquine and chloroquine and sulfadoxine/pyrimethamine. Reported adverse effects were minor or moderate, not life threatening. REVIEWER'S CONCLUSIONS: There is some evidence to support the continued use of amodiaquine in the treatment of uncomplicated malaria, although drug resistance should be considered. Monitoring for toxicity should also continue. PMID- 10796469 TI - Angioplasty (versus non surgical management) for intermittent claudication. AB - BACKGROUND: Intermittent claudication is pain in the legs due to muscle ischaemia associated with arterial stenosis or occlusion. Angioplasty is a technique that involves dilatation and recanalisation of a stenosed or occluded artery. OBJECTIVES: The objective of this review was to determine the effects of angioplasty of arteries in the leg when compared with non surgical therapy, or no therapy, for patients with mild to moderate intermittent claudication. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register and reference lists of relevant articles. The reviewers also contacted investigators in the field and hand searched recent conference proceedings. SELECTION CRITERIA: Randomised trials of angioplasty for mild or moderate intermittent claudication. DATA COLLECTION AND ANALYSIS: One reviewer extracted data and both reviewers assessed trial quality independently. MAIN RESULTS: Two trials with a total of 98 participants were included. The average age was 62 years old with 20 women and 789 men. Patients were followed for 15 months in one trial and six years in another. At six months of follow up, mean ankle brachial pressure indices were higher in the angioplasty groups than control groups (weighted mean difference 0.17, 95% confidence interval 0.11 to 0.24). In one trial, walking distances were greater in the angioplasty group, but in the other trial, in which controls underwent an exercise programme, walking distances did not show a greater improvement in the angioplasty group. At two years of follow up in one trial, the angioplasty group were more likely to have a patent artery (odds ratio 5.5, 95% confidence interval 1.8 to 17.0) but not a significantly better walking distance or quality of life. In the other trial, long term follow up at six years demonstrated no significant differences in outcome between the angioplasty and control groups. REVIEWER'S CONCLUSIONS: These limited results suggest that angioplasty may have had a short term benefit, but this may not have been sustained. PMID- 10796470 TI - Antibiotics for preventing respiratory tract infections in adults receiving intensive care. AB - OBJECTIVES: Pneumonia is an important cause of mortality in intensive care units. The objective of this review was to assess the effects of antibiotics for preventing respiratory tract infections and overall mortality in adults receiving intensive care. SEARCH STRATEGY: We searched MEDLINE, proceedings of scientific meetings and reference lists of articles from January 1984 to September 1997. We also contacted investigators in the field. SELECTION CRITERIA: Randomised trials of antibiotic prophylaxis for respiratory tract infections and deaths among adult intensive care unit patients. DATA COLLECTION AND ANALYSIS: Trials were assessed for quality and investigators contacted for additional information. MAIN RESULTS: Overall 33 trials involving 5727 people were included. There was variation in the antibiotics used, patient characteristics and the risk of respiratory tract infections and mortality in the control groups. In 16 trials (involving 3493 patients) of a topical and systemic antibiotic combination, the average rates of respiratory tract infections and deaths in the control group were 33% and 28% respectively. There was a significant reduction of both respiratory tract infections (odds ratio 0.36, 95% confidence interval 0.30 to 0. 43) and total mortality (odds ratio 0.80, 95% confidence interval 0. 68 to 0.93). On average five patients needed to be treated to prevent one infection and 23 treated to prevent one death. In 17 trials (involving 2366 patients) of topical antimicrobials the rates of respiratory tract infections and deaths in the control groups were 30% and 24% respectively. There was a significant reduction of respiratory tract infections (odds ratio 0.57, 95% confidence interval 0.46 to 0.69) but not in total mortality (odds ratio 1.01, 95% confidence interval 0.84 to 1.22). REVIEWER'S CONCLUSIONS: A combination of topical and systemic prophylactic antibiotics can reduce respiratory tract infections and overall mortality in adult patients receiving intensive care. [This abstract has been prepared centrally.] PMID- 10796471 TI - Antibiotics for sore throat. AB - BACKGROUND: Sore throat is a very common reason for people to attend for medical care. Sore throat is a disease that remits spontaneously, that is, 'cure' is not dependant on treatment. Nonetheless primary care doctors commonly prescribe antibiotics for sore throat and other upper respiratory tract infections. OBJECTIVES: To assess the benefits of antibiotics in the management of sore throat. SEARCH STRATEGY: Systematic search of the literature from 1945 to 1999, using electronic searches of MEDLINE (using the keywords, "pharyngitis", "sore throat" and "tonsillitis") after 1966, the Cochrane Library, the Cochrane collection of hand-searched trials, and the reference sections of the articles identified. Abstracts of identified articles were used to determine which studies were trials. SELECTION CRITERIA: Trials of antibiotic against control with either measures of the typical symptoms (throat soreness, headache or fever), or complications (suppurative and non-suppurative) of sore throat. DATA COLLECTION AND ANALYSIS: RevMan 4.0.3 MAIN RESULTS: A total number of 10,484 cases of sore throat have been studied. 1. Non-suppurative complications There was a trend for protection against acute glomerulonephritis by antibiotics, but insufficient cases were recorded to be sure of this effect. Several studies found benefit from antibiotics for acute rheumatic fever, which reduced this complication to less than one third (OR = 0.30; 95% CI = 0.20-0.45). 2. Suppurative complications Antibiotics reduced the incidence of acute otitis media to about one quarter of that in the placebo group (OR = 0.22; 95% CI = 0.11-0.43) and reduced the incidence of acute sinusitis to about one half of that in the placebo group (OR = 0.46; 95% CI = 0.10-2.05). The incidence of quinsy was also reduced in relation to placebo group (OR = 0.18; 95% CI = 0.08-0.43). 3. Symptoms Symptoms of headache, throat soreness and fever were reduced by antibiotics to about one half. The greatest time for this to be evident was at about three and a half days (when the symptoms of about 50% of untreated patients had settled). About 90% of treated and untreated patients were symptom-free by one week. 4. Subgroup analyses of symptom reduction Subgroup analysis by age; blind vs unblinded; us of antipyretics; or results of swabs for Streptococcus yielded no significant differences. REVIEWER'S CONCLUSIONS: Antibiotics confer relative benefits in the treatment of sore throat. However, the absolute benefits are modest. Protecting sore throat sufferers against suppurative and non-suppurative complications in modern Western society can only be achieved by treating many with antibiotics who will derive no benefit. Antibiotics shorten the duration of symptoms, but by a mean of only about half of one day at day 3 (the time of maximal effect), and by about eight hours overall. PMID- 10796472 TI - Anxiolytics and antidepressants for smoking cessation. AB - BACKGROUND: There are two reasons to believe antidepressants and anxiolytics might help in smoking. First, anxiety and depression are symptoms of nicotine withdrawal, and smoking cessation sometimes precipitates depression. Second, smoking appears to be due, in part, to deficits in dopamine, serotonin and norepinephrine, all of which are increased by anxiolytics and antidepressants. OBJECTIVES: The aim of this review is to assess the effectiveness of such drugs in aiding long term smoking cessation. The drugs include bupropion; buspirone; diazepam; doxepin; fluoxetine; imipramine; meprobamate; moclobemide; nortriptyline; tryptophan; ondansetron; venlafaxine and the beta-blockers metoprolol, oxprenolol and propanolol. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register which includes trials indexed in Medline, Embase, SciSearch and PsycLit, and meetings abstracts. SELECTION CRITERIA: We considered randomized trials comparing anxiolytic or antidepressant drugs to placebo or an alternative therapeutic control for smoking cessation. We excluded trials with less than 6 months follow-up. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of study population, the nature of the drug therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: There was one trial each of the anxiolytics diazepam, meprobamate, metoprolol and oxprenolol. There were two trials of the anxiolytic buspirone. None of these showed evidence of effectiveness in helping smokers to quit. There was one trial each of the antidepressants fluoxetine and moclobemide, two of nortriptyline, and four trials of bupropion. Nortriptyline and bupropion increased cessation and other antidepressants might also be effective. One trial found combined bupropion and nicotine patch produced higher quit rates than patch alone. REVIEWER'S CONCLUSIONS: There is little evidence that anxiolytics aid smoking cessation. Some antidepressants (bupropion and nortriptyline) can aid smoking cessation. It is not clear whether these effects are specific for individual drugs, or a class effect. PMID- 10796473 TI - Bromocriptine for unexplained subfertility in women. AB - BACKGROUND: Bromocriptine improves hyperprolactinemic amenorrhea and so could also be helpful in the treatment of unexplained subfertility in women. OBJECTIVES: To assess the effects of bromocriptine in women with unexplained subfertility. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Randomised trials comparing bromocriptine with placebo or no treatment in women with unexplained subfertility. DATA COLLECTION AND ANALYSIS: Two reviewers applied the eligibility criteria and assessed trial quality independently. MAIN RESULTS: Three trials involving of 127 women were included. All trials were double-blind comparisons with placebo, and one was of crossover design. Conception rates with bromocriptine treatment did not improve compared with placebo (odds ratio was 1.12, 95% confidence interval 0.48 to 2.57). REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate bromocriptine use in women with unexplained subfertility. However trials for women with unexplained subfertility who also have expressible galactorrhea may be worthwhile. PMID- 10796474 TI - Bronchopulmonary hygiene physical therapy for chronic obstructive pulmonary disease and bronchiectasis. AB - BACKGROUND: Bronchopulmonary hygiene physical therapy is a form of chest physical therapy including chest percussion and postural drainage to remove lung secretions. These are applied commonly to patients with both acute and chronic airway diseases. Despite controversies in the literature regarding its efficacy, it remains in use in a variety of clinical settings. The various forms of this therapy are labour intensive and need to be evaluated. OBJECTIVES: The objective of this review was to assess the effects of bronchial hygiene physical therapy in people with chronic obstructive pulmonary disease and bronchiectasis. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles up to July 1997. We also wrote to study authors. SELECTION CRITERIA: Randomised trials in which postural drainage, chest percussion, vibration, chest shaking, directed coughing or forced exhalation technique was compared to other drainage or breathing techniques, placebo or no treatment. DATA COLLECTION AND ANALYSIS: Two reviewers applied the inclusion and exclusion criteria on masked publications independently. They assessed the trial quality independently. Only data from the first arm of crossover trials were included. MAIN RESULTS: The seven included trials involved six comparisons and a total of 126 people. The trials were small and not generally of high quality. The results could not be combined as trials addressed different patient groups and outcomes. In most comparisons, bronchial hygiene physical therapy produced no significant effects on pulmonary function, apart from clearing sputum in chronic obstructive pulmonary disease and in bronchiectasis. REVIEWER'S CONCLUSIONS: There is not enough evidence to support or refute the use of bronchial hygiene physical therapy in people with chronic obstructive pulmonary disease and bronchiectasis. PMID- 10796475 TI - Holding chambers versus nebulisers for beta-agonist treatment of acute asthma. AB - BACKGROUND: In acute asthma inhaled beta-agonists are often administered to relieve bronchospasm by wet nebulisation, but some have argued that metered-dose inhalers with a holding chamber (spacer) can be equally effective. In the community setting nebulisers are more expensive, require a power source and need regular maintenance. OBJECTIVES: There is controversy as to whether wet nebulisers are better than metered dose inhalers with holding chambers to deliver beta2-agonist medications for acute asthma. Comparisons of hospital and home use are also of interest. The objective of this review was to assess the effects of holding chambers compared to nebulisers for the delivery of beta2-agonists for acute asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials in adults and children (from two years of age) with asthma, where holding chamber beta2-agonist delivery was compared with wet nebulisation. DATA COLLECTION AND ANALYSIS: One reviewer applied study inclusion criteria and extracted the data. Trial quality was assessed independently by two reviewers. Missing data were obtained from the authors or estimated. MAIN RESULTS: This review analysed 686 children and 375 adults included in 16 trials. Method of delivery of beta2-agonist did not appear to affect hospital admission rates. In adults, the odds ratio of holding chamber versus nebuliser was 1.12, 95% confidence interval 0.45 to 2.76. The odds ratio for children was 0.91, 95% confidence interval 0.4 to 2.1. Children's length of stay in the emergency department was significantly shorter when the holding chamber was used, with a weighted mean difference of -0.62 hours, 95% confidence interval -0.84 to -0.40 hours. Adults' length of stay in the emergency department was similar for the two delivery methods. Peak flow and forced expiratory volume were also similar for the two delivery methods. Pulse rate was lower for holding chamber in children, weighted mean difference -8.3% baseline, 95% confidence interval -11.5 to -5.0,. REVIEWER'S CONCLUSIONS: Metered-dose inhalers with holding chamber produced outcomes that were at least equivalent to nebuliser delivery. Holding chambers may have some advantages compared to nebulisers for children with acute asthma. PMID- 10796476 TI - Interventions for treating schistosomiasis haematobium. AB - BACKGROUND: Schistosomiasis is a parasite that is carried by freshwater snails. There are two common forms, urinary schistosomiasis (which is considered in this review) and intestinal schistosomiasis. OBJECTIVES: The objective of this review was to assess the effects of drugs for treatment of Schistosomiasis haematobium. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, Medline and reference lists of articles were searched. The WHO Division of Control of Tropical Diseases was contacted. SELECTION CRITERIA: Randomised trials of metrifonate or praziquantel or other drugs for treating Schistosomiasis haematobium. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data, and this was checked by a review editor. MAIN RESULTS: Five trials, all from Africa, were included. The quality of the trials was variable. There were no good randomised controlled trials of praziquantel single dose treatment versus current standard treatment with metrifonate of three doses of 10 milligrams per kilogram at two weekly intervals. Praziquantel at doses of 40 milligram per kilogram was more effective than single dose metrifonate 10 milligrams per kilogram (odds ratio 6.94, 95% confidence interval 4.85 to 9.92). In one trial of metrifonate compared with praziquantel, there was no difference demonstrated in a range of clinical outcomes including cessation of haematuria and proteinuria. Both drugs improved nutritional status and physical fitness. REVIEWER'S CONCLUSIONS: Praziquantel (single dose) appears to be more effective than metrifonate (split dose) in terms of parasitological cure of Schistosomiasis haematobium, but the reinfection rate is high with both drugs. PMID- 10796477 TI - Clomiphene citrate for ovulation induction in women with oligo-amenorrhoea. AB - BACKGROUND: The administration of clomiphene citrate is followed by an enhanced release of pituitary gonadotropins resulting in follicular recruitment. After the drug is stopped, there is continuing secretion of estradiol, selection of the dominant follicle and, in successful cases, ovulation. Clomiphene is indicated as the initial treatment in the majority of women with amenorrhoea and oligomennorhoea. In women with irregular ovulation it seems to re-establish typical frequency of ovulation. Its effectiveness in oligo-amenorrhoeic women was tested in a number of randomized controlled trials at that time. These trials form the basis for the following review. OBJECTIVES: Clomiphene citrate enhances the release of pituitary hormones, often resulting in ovulation. The objective of this review was to assess the effects of clomiphene citrate on ovulation and pregnancy in women with oligo-ovulatory subfertility. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Randomised trials of clomiphene compared with placebo or no treatment in women with oligo-ovulatory subfertility of at least 12 months duration. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Four studies were included. They were all of crossover design. Since it was not possible to separate data from the first and second phases of these trials, the effect of clomiphene may be overestimated. Compared with placebo, clomiphene citrate was associated with increased ovulation. The odds ratio for high doses (50-250 milligrams per day) was 6.82 (95% confidence interval 3.92 to 11.85). This dropped to a non-significant odds ratio of 1.29 (95% confidence interval 0.48 to 3.49) with low doses (10 milligrams per day). Clomiphene citrate (all doses) was associated with an increased pregnancy rate per treatment cycle (odds ratio 3.41, 95% confidence interval 4.23 to 9.48). REVIEWER'S CONCLUSIONS: Clomiphene citrate (at doses between 50 to 250 milligrams per day) appears to be an effective method of inducing ovulation and improving fertility in oligo-ovulatory women. However adverse effects include possible ovarian cancer risk and risk of multiple pregnancy. PMID- 10796478 TI - Clomiphene citrate for unexplained subfertility in women. AB - OBJECTIVES: Clomiphene citrate appears to increase ovulation in women with oligo ovulatory subfertility. It may also work in women with unexplained subfertility, perhaps by correcting an unidentifiable ovulatory dysfunction. The objective of this review was to assess the effects of clomiphene citrate in women with unexplained subfertility. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched". SELECTION CRITERIA: Randomised trials of clomiphene citrate (doses of 50 to 250 milligrams per day up to 10 days) compared to placebo or no treatment in women with unexplained subfertility. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. MAIN RESULTS: Five studies were included. Four trials were of crossover design, and quality of the randomisation was variable. Compared to placebo, clomiphene citrate was associated with an increase in pregnancy rates. The odds ratio for pregnancy per patient was 2.38 (95% confidence interval 1.22 to 4.62). The odds ratio of pregnancy per cycle was 2.5 (95% confidence interval 1.35 to 4.62). REVIEWER'S CONCLUSIONS: Clomiphene citrate appears to modestly improve pregnancy rates in women with unexplained subfertility. However adverse effects include a possible ovarian cancer risk and risk of multiple pregnancy. PMID- 10796479 TI - Clonidine for smoking cessation. AB - BACKGROUND: Clonidine was originally used to lower blood pressure. It acts on the central nervous system and may reduce withdrawal symptoms in various addictive behaviours, including tobacco use. OBJECTIVES: The aim of this review is to determine clonidine's effectiveness in helping smokers to quit. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. Date of the most recent search: October 1998. SELECTION CRITERIA: We considered randomised trials of clonidine versus placebo with a smoking cessation endpoint assessed at least 12 weeks following the end of treatment. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the dose and duration of clonidine therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least twelve weeks follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: Six trials met the inclusion criteria. There were three trials of oral, and three of transdermal clonidine. Some form of behavioural counselling was offered to all participants in five of the six trials. There was a statistically significant effect of clonidine in one of these trials. The pooled odds ratio for success with clonidine vs placebo was 1.89 (95% confidence interval 1.30 to 2.74). There was a high incidence of dose-dependent side-effects, particularly dry mouth and sedation. REVIEWER'S CONCLUSIONS: Based on a small number of trials, in which there are potential sources of bias, clonidine is effective in promoting smoking cessation. Prominent side-effects limit the usefulness of clonidine for smoking cessation. PMID- 10796480 TI - Combined inhaled anticholinergic agents and beta-2-agonists for initial treatment of acute asthma in children. AB - BACKGROUND: Anti-cholinergic agents and beta2-agonist drugs are both bronchodilators used to reverse acute bronchospasm in children with asthma. These drugs have different modes of action, so may have complementary or additive effects. OBJECTIVES: The objective of this review was to assess the effects of adding inhaled anti-cholinergics to beta2-agonists in acute paediatric asthma. SEARCH STRATEGY: We searched Medline (1966 to 1996), Embase (1980 to 1995), Cinahl (1982 to 1995) and reference lists of studies. We also contacted drug manufacturers and researchers. SELECTION CRITERIA: Randomised trials comparing the combination of inhaled anti-cholinergics and beta2-agonists with beta2 agonists alone in children aged 18 months to 17 years with acute asthma. DATA COLLECTION AND ANALYSIS: Assessments of trial quality and data extraction were done by two reviewers independently. MAIN RESULTS: Ten trials involving a total of 836 children were included. Most trials were of high quality. When only one dose of anti-cholinergic inhalation was added to beta2-agonist therapy, there was an improvement in forced expiratory volume in one second after 60 minutes with combination therapy (weighted mean difference 16.1%, 95% confidence interval 5.5 to 26. 7% reduction). There was no reduction in hospital admission (odds ratio 0.80, 95% confidence interval 0.35 to 1.82, using a random effects model). For multiple doses in children with severe asthma, there was a reduction in forced expiratory volume in 1 second (weighted mean difference 9.8% predicted, 95% confidence interval 6. 5 to 13.1% predicted). There may also be a reduction in hospital admission (odds ratio 0.62, 95% confidence interval 0.38 to 0.99). Eleven children would need to be given multiple doses of anti-cholinergics in combination with beta2-agonists to avoid one hospital admission compared to children given beta2-agonists alone. REVIEWER'S CONCLUSIONS: In children with acute asthma, the addition of multiple doses of anti-cholinergics to inhaled beta2-agonists appears to improve lung function modestly and may decrease hospital admission. There is no associated increase in adverse effects. Single doses of anti-cholinergics may improve lung function in children with severe asthma, but do not appear to reduce hospital admissions. PMID- 10796481 TI - Follicle-stimulating hormone and human menopausal gonadotropin for ovarian stimulation in assisted reproduction cycles. AB - OBJECTIVES: To conduct a systematic overview of available data comparing FSH and hMG in IVF treatment cycles. SEARCH STRATEGY: This review has drawn on the search strategy developed for the Subfertility Group as a whole. Relevant trials were identified in the Group's Specialised Register of Controlled Trials. See Review Group details for more information. DATA COLLECTION AND ANALYSIS: A systematic review and meta-analysis of randomized trials of FSH versus hMG use in ovarian stimulation protocols, with or without GnRH agonists, in IVF treatment cycles. Common odds ratios (OR) were calculated after demonstrating homogeneity of treatment effect across all trials. MAIN RESULTS: MAIN OUTCOME MEASURES: Clinical pregnancy rates per cycle started, per cycle reaching oocyte retrieval, and per cycle reaching embryo transfer (ET). RESULTS: Eight trials met the inclusion criteria. The overall OR in favour of FSH for cycle start, oocyte retrieval, and ET were 1.70 (95% CI, 1.11-2.60), 1.68 (95% CI, 1.10-2.56), and 1.69 (95% CI, 1.10-2.59), respectively. REVIEWER'S CONCLUSIONS: This meta analysis demonstrates that in IVF cycles the use of FSH is associated with a significantly higher clinical pregnancy rate than hMG. PMID- 10796482 TI - Azathioprine for maintaining remission of Crohn's disease. AB - OBJECTIVES: To assess the effectiveness of azathioprine in maintaining remission of quiescent Crohn's disease. SEARCH STRATEGY: Pertinent studies were selected using the MEDLINE data base (1966 - May 1998), the Cochrane Controlled Trials Register, the Inflammatory Bowel Disease Trials Register, as well as abstracts from major gastrointestinal research meetings and references from published articles and reviews. SELECTION CRITERIA: Five randomized, double-blind, placebo controlled trials of azathioprine therapy were identified. Two of these trials consisted solely of patients with quiescent Crohn's disease. Three trials had multiple therapeutic arms for both induction of remission and maintenance of remission. DATA COLLECTION AND ANALYSIS: Data were extracted by three independent observers (GRM, GF, LRS) based on the intention to treat principle. Peto odds ratios for the overall maintenance of remission, steroid sparing, and withdrawals due to adverse effects were calculated, and from these, 95% confidence intervals were derived. Numbers needed to treat or harm (NNT, NNH respectively) for the maintenance of remission, steroid sparing, and withdrawals due to adverse effects were also determined. MAIN RESULTS: Azathioprine had a positive effect on maintaining remission. The Peto odds ratio for maintenance of remission was 2.16 (CI 1.35 - 3.47) with an NNT of 7. A higher dose improved response. A steroid sparing effect was noted, with a Peto odds ratio of 5.22 (CI 1.06 - 25.68) and NNT of 3 for quiescent disease. The Peto odds ratio for withdrawals due to adverse events was 4.36 (CI 1.63 - 11.67), the NNH (Number Needed to Harm) was 19. REVIEWER'S CONCLUSIONS: Azathioprine is effective in maintaining remission. There is evidence for a steroid sparing effect. PMID- 10796483 TI - Danazol for pelvic pain associated with endometriosis. AB - BACKGROUND: Endometriosis is defined as the presence of endometrial tissue (stromal and glandular) outside the normal uterine cavity. Conventional medical and surgical treatments for endometriosis aim to remove or decrease deposits of ectopic endometrium. The observation that hyperandrogenic states (an excess of male hormone) induce atrophy of the endometrium has led to the use of androgens in the treatment of endometriosis. Danazol is one of these treatments used. The efficacy of danazol is based on its ability to produce a high androgen/low estrogen environment (a pseudo menopause) which results in the atrophy of endometriotic implants and thus an improvement in painful symptoms. OBJECTIVES: To determine the effectiveness of danazol compared to placebo or no treatment in the treatment of the symptoms and signs, other than infertility, of endometriosis in women of reproductive age. SEARCH STRATEGY: The Menstrual Disorders Group search strategy was used to identify randomised controlled trials of the use of danazol in endometriosis. In addition, all reference lists of included trials were searched, and relevant drug companies were contacted for details of unpublished trials SELECTION CRITERIA: Randomised controlled trials in which danazol (alone or as adjunctive therapy) was compared to placebo or no therapy. Trials which only reported infertility outcomes were excluded. DATA COLLECTION AND ANALYSIS: Only four trials met the inclusion criteria and two authors extracted data independently from these trials. All four trials compared danazol to placebo. Two trials used danazol as sole therapy and two trials used danazol as an adjunct to surgery. Although the main outcome was pain improvement other data relating to laparoscopic scores and hormonal parameters were also collected. MAIN RESULTS: Treatment with danazol (including adjunctive surgical therapy) was effective in relieving painful symptoms related to endometriosis when compared to placebo. Laparoscopic scores were improved with danazol treatment (including adjunctive therapy) when compared with either placebo or no treatment. Side effects were more commonly reported in those patients receiving danazol than placebo. REVIEWER'S CONCLUSIONS: Danazol is effective in treating the symptoms and signs of endometriosis. However, its use is limited by the occurrence of androgenic side effects. PMID- 10796484 TI - Danazol for unexplained infertilty. AB - BACKGROUND: The androgen, Danazol, was developed in the 1970's as a treatment for endometriosis. Its use was soon advocated in women with unexplained infertility. Two randomized trials were subsequently conducted to assess the effectiveness of danazol in this population. OBJECTIVES: The objective of this review was to assess the effects of danazol on pregnancy rates in women with unexplained subfertility. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Randomised trials of danazol compared with placebo or no treatment in women with unexplained subfertility. DATA COLLECTION AND ANALYSIS: Data were extracted by two reviewers. MAIN RESULTS: Two trials involving 68 women were involved. There was no difference found in pregnancy rate between danazol and placebo (odds ratio 2.57, 95% confidence 0.53 to 12.46). REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the effect of danazol on pregnancy rates in women with unexplained subfertility. The need to use contraception during danazol treatment, adverse effects and costs are additional considerations. PMID- 10796485 TI - Interventions to promote collaboration between nurses and doctors. AB - BACKGROUND: Lack of nurse-doctor collaboration contributes to problems in quality and efficiency of patient care. OBJECTIVES: To assess the effects of interventions designed to improve nurse-doctor collaboration. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register and database of studies awaiting assessment, the Cochrane Database of Systematic Reviews, the Cochrane Controlled Trials Register, the Database of Abstracts of Reviews of Effectiveness, MEDLINE, and reference lists of articles up to the end of October 1999. SELECTION CRITERIA: Randomised trials, controlled before-and-after studies and interrupted time series of interventions to improve collaboration between nursing and medical professionals sharing patient care in primary or hospital care settings. DATA COLLECTION AND ANALYSIS: One reviewer assessed the eligibility of potentially relevant studies, extracted data and assessed the quality of included studies; a second reviewer undertook duplicate assessments on the eligibility of some articles and data abstraction on all included studies. MAIN RESULTS: Two trials involving 1945 people were included. One six month trial involving 1102 admissions evaluated daily, structured, team ward rounds, in which nurses, doctors and other professionals made care decisions jointly. There was shortened average length of hospital stay (LOS) from 6.06 to 5.46 days, and reduced hospital charges from US$ 8090 to 6681. There were no differences in mortality rates or the type of care to which patients were discharged. Another three month trial involving 843 admissions compared two female wards and evaluated a four times per week round. There were no significant differences between the intervention and control wards in total average length of stay for all patients (11.7 days in intervention ward versus 11.6 in the control ward). Excluding patients who died in hospital revealed shortened length of stay in the intervention ward (intervention ward 10.5 days, control ward 11.9). Mortality rates were not significantly different. REVIEWER'S CONCLUSIONS: Increasing collaboration improved outcomes of importance to patients and to health care managers. These gains were moderate and affected health care processes rather than outcomes. Further research is needed to confirm these findings. The logistic challenge presented by the complexity of the interventions and the need for large sample sizes due to the likely modest impact and rarity of outcome events may best be met by multi-centre studies. Before launching such studies qualitative research is needed to identify barriers to collaboration. Interventions other than nurse-doctor ward rounds and team meetings should also be tested. PMID- 10796486 TI - Family therapy for asthma in children. AB - BACKGROUND: Psychosocial and emotional factors are important in childhood asthma. Nevertheless, drug therapy alone continues to be the main treatment. Treatment programmes that include behavioural or psychological interventions have been developed to improve disturbed family relations in the families of children with severe asthma. These approaches have been extended to examine the efficacy of family therapy to treat childhood asthma in a wider group of patients. This review systematically examines these studies. OBJECTIVES: Recognition that asthma can be associated with emotional disturbances has led to the investigation of the role of family therapy in reducing the symptoms and impact of asthma in children. The objective of this review was to assess the effects of family therapy as an adjunct to medication for the treatment of asthma in children. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, Psychlit and Psychinfo. SELECTION CRITERIA: Randomised trials comparing children undergoing systematic therapy focusing on the family in conjunction with asthma medication, with children taking asthma medication only. DATA COLLECTION AND ANALYSIS: One reviewer applied the study inclusion criteria. MAIN RESULTS: Two trials with a total of 55 children were included. It was not possible to combine the findings of these two studies because of differences in outcome measures used. In one study, gas volume, peak expiratory flow rate and daytime wheeze showed improvement in family therapy patients compared to controls. In the other study, there was an improvement in overall clinical assessment and number of functionally impaired days in the patients receiving family therapy. There was no difference in forced expiratory volume or medication use in both studies. REVIEWER'S CONCLUSIONS: There is some indication that family therapy may be a useful adjunct to medication for children with asthma. This conclusion is limited by small study sizes and lack of standardisation in the choice of outcome measures. PMID- 10796487 TI - Garlic for peripheral arterial occlusive disease. AB - BACKGROUND: Commercially available preparations of garlic have been reported to have beneficial effects on some of the risk factors associated with atherosclerosis. OBJECTIVES: The objective of this review was to assess the effects of garlic (both dried and non-powdered preparations) for the treatment of peripheral arterial occlusive disease. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register, AMED, EMBASE, BIDS ISI, abstracts of relevant symposia and reference lists of relevant articles up to August 1998. The reviewers also contacted pharmaceutical companies, investigators and experts in garlic therapies. SELECTION CRITERIA: Randomised trials of garlic therapy in patients with lower limb atherosclerosis. The main outcomes were objective measures of progression of underlying atherosclerosis (e.g. ankle pressure measurements, treadmill testing) and subjective measures (e.g. symptom progression). DATA COLLECTION AND ANALYSIS: At least two reviewers extracted data and assessed trial quality independently. The reviewers contacted investigators to obtain information needed for the review that could not be found in published reports. MAIN RESULTS: One eligible trial with 78 participants was found. Both men and women (aged 40-75) were included. The follow-up period was short, 12 weeks only. After twelve weeks of treatment, pain free walking distance increased from 161 to 207 m in the group on garlic and from 172 to 203 m in the placebo group. This was not a statistically significant difference. There was no difference in change of systolic or diastolic blood pressure, heart rate, ankle and brachial pressures. No severe side effects were observed and nine patients taking garlic (28%) and four patients taking placebo (12%) complained of a noticeable garlic smell. REVIEWER'S CONCLUSIONS: One small trial of short duration found no effect on walking distance. PMID- 10796488 TI - Gonadotrophin-releasing hormone analogue as an adjunct to gonadotropin therapy for clomiphene-resistant polycystic ovarian syndrome. AB - BACKGROUND: Elevation of endogenous LH levels may result in premature luteinization. This may also be associated with the increased rate of spontaneous abortion. Gonadotropin releasing hormone analogue (GnRHa) used prior to human menopausal gonadotropin (hMG/FSH) administration may improve the outcome of ovulation induction. OBJECTIVES: To assess if GnRHa pre-treatment plus FSH/hMG increase the rate of clinical pregnancy and/or decrease the rate of spontaneous abortion in women with WHO group 2 ovulatory dysfunction, compared with hMG/FSH alone. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: All relevant published and unpublished RCTs were selected. Three RCTs were identified comparing these two approaches. DATA COLLECTION AND ANALYSIS: DATA EXTRACTION: A diverse search strategy was employed, including hand-search of 43 core journals from 1966 to the present, bibliographies of relevant trials, MEDLINE database, abstracts from North American and European meetings and contact with authors of relevant papers. Relevant data were extracted independently by two reviewers using the standardised data extraction sheet. Validity was assessed in terms of method of randomisation, completeness of follow-up, presence or absence of crossover and co-intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were generated using the Peto modified Mantel Haenszel technique. Statistical heterogeneity was assessed using x2. MAIN RESULTS: Studies were clinically and statistically homogenous. Common odds ratios for pregnancy per treatment cycle and moderate to severe ovarian hyperstimulation syndrome (OHSS) were 1.50 (0.72-3.12) and 1.40 (0.5-3.92) respectively. REVIEWER'S CONCLUSIONS: These studies are too small to clearly demonstrate clinically significant differences in pregnancy rate between the two approaches. However, data from IVF studies suggest that there may be an increased risk of OHSS associated with GnRHa use. In the absence of evidence suggesting a benefit of GnRHa augmentation for PCOS, it should not be recommended as a standard treatment for this patient group. Further studies assessing live birth and OHSS rates are warranted. PMID- 10796489 TI - Lipid-lowering for lower limb atherosclerosis. AB - BACKGROUND: Raised lipid levels, including cholesterol, are important risk factors in the development of lower limb arterial disease (atherosclerosis). OBJECTIVES: The objective of this review was to assess the effects of lipid lowering therapy in patients with lower limb atherosclerosis. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register, Embase, reference lists of relevant articles, and contacted trial investigators in Europe and pharmaceutical companies. SELECTION CRITERIA: Randomised trials of lipid-lowering therapy in patients with lower limb atherosclerosis. The main outcomes were mortality, non-fatal events, direct tests of disease progression, indirect measurements of disease, and subjective measures. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data and assessed trial quality independently. The reviewers contacted investigators to obtain information needed for the review that could not be found in published reports. MAIN RESULTS: There were nine eligible trials, but two were excluded because of poor methodology. The seven remaining trials involved a total of 698 participants from seven different countries. Men and women participated in all but one trial and were generally middle aged to elderly. The follow-up period varied from four months to three years. The overall quality of the included trials was high. The trials were heterogeneous in terms of inclusion criteria, type of drugs used and outcomes measured. Lipid-lowering therapy produced a marked but non-significant reduction in mortality (odds ratio 0.21, 95% confidence interval 0. 03 to 1.17), but little change in non-fatal events (odds ratio 1.21, 95% confidence interval 0.80 to 1.83). In two trials there was a significant overall reduction in disease progression on angiogram (odds ratio 0.47, 95% confidence interval 0.29 to 0.77). The changes in ankle brachial pressure index and walking distance were inconsistent, although trials showed a general improvement in symptoms that could not be combined in a meta-analysis. Side effects were generally mild, with the exceptions of liver toxicity produced by betapyridil and the adverse effect of probucol on lipoprotein profile. REVIEWER'S CONCLUSIONS: Lipid-lowering therapy may be useful in preventing deterioration of underlying disease and alleviating symptoms. These results cannot determine whether one lipid-lowering regimen is better than another. PMID- 10796490 TI - Lobeline for smoking cessation. AB - BACKGROUND: Lobeline is a partial nicotine agonist, which has been used in a variety of commercially available preparations to help stop smoking. OBJECTIVES: The objective of this review was to assess the effects of lobeline on long term smoking cessation. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: Randomized trials comparing lobeline to placebo or an alternative therapeutic control, which reported smoking cessation with at least six months follow-up. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the dose and form of lobeline, the outcome measures, method of randomisation, and completeness of follow-up. MAIN RESULTS: We identified no trials meeting the full inclusion criteria including long term follow-up. REVIEWER'S CONCLUSIONS: There is no evidence available from long term trials that lobeline can aid smoking cessation. PMID- 10796491 TI - Local opinion leaders: effects on professional practice and health care outcomes. AB - BACKGROUND: Both the theory of diffusion of innovations and the social influences model of behaviour change suggest that using local opinion leaders to transmit norms and model appropriate behaviour may improve health professional practice. OBJECTIVES: To assess the effects using local opinion leaders on the practice of health professionals or patient outcomes. SEARCH STRATEGY: We searched MEDLINE to May 1998, the Research and Development Resource Base in Continuing Medical Education, and reference lists of related systematic reviews and articles. SELECTION CRITERIA: Randomised trials of the use of local opinion leaders (defined as health professionals nominated by their colleagues as being educationally influential). The participants were health care professionals responsible for patient care. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eight studies were included involving more than 296 health professionals. A variety of patient problems were targeted, including acute myocardial infarction, cancer pain, osteoarthritis, rheumatoid arthritis, chronic lung disease, vaginal birth after caesarean section, labour and delivery, and urinary catheter care. Six of seven trials that measured health professional practice demonstrated some improvement for at least one outcome variable, and in two trials, the results were statistically significant and clinically important. In three trials that measured patient outcomes, only one achieved an impact upon practice that was of practical importance: local opinion leaders were effective in improving the rate of vaginal birth after previous caesarean section. REVIEWER'S CONCLUSIONS: Using local opinion leaders results in mixed effects on professional practice. However, it is not always clear what local opinion leaders do and replicable descriptions are needed. Further research is required to determine if opinion leaders can be identified and in which circumstances they are likely to influence the practice of their peers. PMID- 10796492 TI - Vaccines for preventing malaria. AB - BACKGROUND: Despite continued efforts to control the disease, malaria remains a major health problem in many regions of the world, especially sub-Saharan Africa, and new ways to control or eradicate the disease are urgently needed. Two types of vaccine, SPf66 vaccine against the asexual stages, and NANP vaccines against the sporozoite stages of the Plasmodium parasite, have been tested in randomised clinical trials in endemic areas. OBJECTIVES: To assess the effects of malaria vaccines. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles were searched. Organisations and researchers in the field were contacted. SELECTION CRITERIA: Randomised trials comparing vaccines against Plasmodium falciparum, P. vivax, P. malariae or P. ovale, and placebo. DATA COLLECTION AND ANALYSIS: Two independent reviewers assessed trial quality and conducted data extraction. MAIN RESULTS: Thirteen efficacy trials involving about 7700 people were included. There were nine trials of the Spf66 vaccine and four trials of the NANP vaccines. There was large heterogeneity between trials when investigating the effect of SPf66 in reducing incidence of the first attack of P. falciparum malaria. When trials were subcategorised by location, there was no evidence for effect of SPf66 in reducing incidence of P. falciparum in four African trials conducted in children under 5 years of age (Peto odds ratio [OR] = 0.96, 95% confidence interval [CI] 0.81 to 1.14). In five trials outside Africa with participants aged 2 years to adult, there was a reduction in incidence by SPf66 vaccine (Peto OR = 0.77, 95% CI 0.67 to 0.88, fixed effects model). Significant heterogeneity remained between trials conducted outside Africa. Using a random effects model for these five trials, the OR was 0.74 (95% CI 0.54 to 1.01). In five trials, there was no evidence for effect of the SPf66 vaccine on the incidence of the first attack of P. vivax malaria (OR 1.01, 95% CI 0.87 to 1.17). Trials to date have not indicated any severe adverse effects of SPf66 vaccine. In three trials of NANP-based vaccines, there was no evidence for protection by these vaccines against P. falciparum malaria (OR 1.12, 95% CI 0.64 to 1.93). REVIEWER'S CONCLUSIONS: There is no evidence for protection by SPf66 vaccines against P. falciparum in Africa. There is a modest reduction in attacks of P. falciparum malaria following vaccination with SPf66 in other regions. Further research with SPf66 vaccines in South America may be justified. Trials to date have not been of sufficient size to evaluate the effect of malaria vaccines on mortality or on severe malaria requiring admission to hospital. There was not enough evidence to evaluate the use of NANP vaccines. PMID- 10796493 TI - Mefloquine for preventing malaria in non-immune adult travellers. AB - OBJECTIVES: Mefloquine has now largely replaced earlier malaria prophylaxis drugs which are no longer considered to be effective against all Plasmodium species, due to parasite resistance. However mefloquine may be associated with neuropsychological harmful effects. The objective of this review was to assess the effects of mefloquine in adult travellers. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, Medline, Embase, Lilacs, Science Citation Index and reference lists of articles. We contacted researchers in the subject of malaria chemoprophylaxis, and drug companies. SELECTION CRITERIA: Randomised trials comparing mefloquine with other standard prophylaxis or placebo in non-immune adult travellers. DATA COLLECTION AND ANALYSIS: The two reviewers independently assessed trial quality and extracted data. Study authors were also contacted. MAIN RESULTS: Ten trials involving 2750 non-immune adult travellers were included. One trial comparing mefloquine with placebo showed mefloquine prevented malaria episodes in an area of drug resistance (odds ratio 0.04, 95% confidence interval 0.02 to 0.08). Withdrawals in the mefloquine group were consistently higher in four placebo controlled trials (odds ratio 3. 56, 95% confidence interval 1.67 to 7.60). In five trials comparing mefloquine with other chemoprophylaxis, no difference in tolerability was detected. REVIEWER'S CONCLUSIONS: Mefloquine prevents malaria, but there is not enough evidence to evaluate its tolerability and toxicity, particularly for general travellers. PMID- 10796494 TI - Nicotine replacement therapy for smoking cessation. AB - BACKGROUND: The aim of nicotine replacement therapy (NRT) is to replace nicotine from cigarettes. This reduces withdrawal symptoms associated with smoking cessation to help resist the urge to smoke cigarettes. OBJECTIVES: The aims of this review were to determine the effectiveness of the different forms of nicotine replacement therapy (chewing gum, transdermal patches, nasal spray, inhalers and tablets) in achieving abstinence from cigarettes; to determine whether the effect is influenced by the clinical setting in which the smoker is recruited and treated, the dosage and form of the NRT used, or the intensity of additional advice and support offered to the smoker; and to determine whether combinations of NRT are more effective than one type alone. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: Randomized trials in which NRT was compared to placebo or no treatment, or where different doses of NRT were compared. We excluded trials which did not report cessation rates, and those with follow-up of less than six months. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the dose and duration and form of nicotine therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months of follow-up. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: We identified 49 trials of nicotine gum, 32 of transdermal nicotine patch, four of intranasal nicotine spray, four of inhaled nicotine and two of nicotine sublingual tablet. Three trials compared combinations of two forms of nicotine therapy with one form alone. The odds ratio for abstinence with NRT compared to control was 1.72 (95% confidence interval 1.60 to 1.84), The odds ratios for the different forms of NRT were 1.63 for gum, 1.77 for patches, 2.27 for nasal spray, 2.08 for inhaled nicotine and 1.73 for nicotine sublingual tablet. These odds were largely independent of the intensity of additional support provided or the setting in which the NRT was offered. Eight weeks of patch therapy was as effective as longer courses and there was no evidence that tapered therapy was better than abrupt withdrawal. Wearing the patch only during waking hours (16 hours/day) was as effective as wearing it for 24 hours/day. The odds ratio for abstinence in the trials which directly compared 4 mg versus 2 mg gum in highly dependent smokers found a significant benefit in favour of 4 mg gum (odds ratio 2.67, 95% confidence interval 1.69 to 4.22). There is no strong evidence that combinations of forms of NRT are more effective. Only one study directly compared NRT against another pharmacotherapy (bupropion) and found that the latter was significantly more effective either alone or used in combination with nicotine patch than if nicotine patch was used alone. REVIEWER'S CONCLUSIONS: All of the commercially available forms of NRT (nicotine gum, transdermal patch, and in some countries, the nicotine nasal spray, nicotine inhaler and nicotine sublingual tablets) are effective as part of a strategy to promote smoking cessation. They increase quit rates approximately 1.5 to 2 fold regardless of setting. The effectiveness of NRT appears to be largely independent of the intensity of additional support provided to the smoker. Since all the trials of NRT reported so far have included at least some form of brief advice to the smoker, this represents the minimum which should be offered in order to ensure its effectiveness. Provision of more intense levels of support, although beneficial in facilitating the likelihood of quitting, is not essential to the success of NRT. There is promising evidence that bupropion may be more effective than NRT (either alone or in combination). (ABSTRACT PMID- 10796495 TI - Nimodipine for primary degenerative, mixed and vascular dementia. AB - BACKGROUND: Dementia is an age-related condition in which Alzheimer's disease (AD) and cerebrovascular disease account for the bulk of cases. The role played by calcium in regulating brain functions is well known - the calcium ion links membrane excitation to subsequent intracellular enzymatic response. Change in calcium homeostasis is one important effect of aging with repercussions on higher cortical functions. Nimodipine is an isopropyl calcium channel blocker which can easily cross the blood brain barrier. Its primary action is to reduce the number of open channels, thus restricting influx of calcium ions into the cell. The usefulness of nimodipine in patients with Alzheimer's disease and vascular dementia and unspecified dementia is still controversial with mixed results. In spite of the uncertainties about its efficacy in dementia, nimodipine is currently a frequently prescribed drug for cognitive impairment and dementia in several European countries. This review will be conducted in two phases; the current review is based on evidence from published data only. The second phase will be based on individual-patient data analysed centrally and added to this review in due course. OBJECTIVES: To determine the clinical efficacy of nimodipine for the symptoms of dementia, either unclassified or according to the major subtypes - Alzheimer's disease, vascular, or mixed Alzheimer's and vascular dementia. SEARCH STRATEGY: The Cochrane Dementia Group Register of Clinical Trials was searched using the terms 'nimodipine' and 'isopropyl (2-methoxy-ethyl) 1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3, 5-pyridinedicarboxylate'. SELECTION CRITERIA: All unconfounded, double-blind, randomised trials in which treatment with nimodipine was administered for more than a day and compared to placebo in patients with dementia, either unclassified or according to the major subtypes - Alzheimer's disease, vascular, or mixed Alzheimer's and vascular dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers and the odds ratio (95%CI) or the average difference (95%CI) were estimated. Both intention-to-treat and on-treatment results were extracted. MAIN RESULTS: This review produced no clear results. Many of the data published were not capable of being sensibly pooled. The data were compatible with nimodipine producing improvement, no change or even harm for those with Alzheimer's disease, vascular dementia, or mixed Alzheimer's and vascular dementia. It was not possible to use many of the published results in a combined analysis. For measures of overall clinical improvement, the intention-to-treat analysis, based on one study only, failed to detect any difference between nimodipine and placebo (OR 0.53; 95%CI 0.25 - 1.13). An on-treatment analysis, based on one study only, produced a statistically significant difference in favour of nimodipine (SMD 4.4; 95%CI 3.9 - 5.0). For cognitive function, the effect of nimodipine was statistically significantly different from placebo for the Mini Mental State Examination score (0-30; high =good) (SMD 0.9; 95%CI 0.59 - 1.22) and there was a statistically significant effect in favour of treatment for the Wechsler Memory Scale (SMD 0.47; 95%CI 0.17 - 0.77). These analyses were based only on those who completed the study and not intention-to-treat analyses. There were no results presented in a form suitable for pooling for functional autonomy, behaviour, quality of life dependency (eg institutionalization), effect on carer, death, acceptability of treatment (as measured by withdrawal rate, safety (as measured by the incidence of adverse effects, including side effects, leading to withdrawal). REVIEWER'S CONCLUSIONS: This review provides no convincing evidence that nimodipine is a useful treatment for the symptoms of dementia, either unclassified or according to the major subtypes - Alzheimer's disease, vascular, or mixed Alzheimer's and vascular dementia. (ABSTRACT TRUNCATED) PMID- 10796496 TI - Androgens versus placebo or no treatment for idiopathic oligo/asthenospermia. AB - BACKGROUND: Oligo-astheno-teratospermia (sperm of low concentration, reduced motility and increased abnormal morphology) of unknown cause is common and the need for treatment is felt by patients and doctors alike. As a result, a variety of empirical, non-specific treatments have been used in an attempt to improve semen characteristics and fertility. Androgens have been suggested as a treatment because its binding proteins maintain a maintain a high intratesticular level testosterone essential for spermatogenesis and because the epididymis and seminal vesicles affect the seminal constitution and sperm motility and are also androgen dependent. However exogenous testosterone was found to exert negative feedback on the pituitary-gonadal axis and thereby to suppress FSH and LH secretion. Spermatogenesis was thus adversely affected. Nevertheless androgens are used for the treatment of male infertility either for a putative direct "stimulatory" or "rebound" therapy. The stimulatory androgens used are mesterolone and testosterone undecanoate which, it is postulated, in a form and dosage that does not influence pituitary gonadotrophin secretion, either have a direct stimulatory effect on spermatogenesis or influence sperm transport and maturation though an effect on the epididymis, ductus deferens and seminal vesicles. Other androgens have been used to produce a rebound effect. These androgens are administered to suppress gonadotrophin secretion and spermatogenesis. After androgen therapy is discontinued there is a surge of FSH and LH and spermatogenesis is recommenced. Because of their different proposed mechanisms of action, stimulatory and rebound androgen therapy are analysed separately in the comparisons. This review considers the available evidence of the effect of androgens for idiopathic oligo and/or asthenospermia. OBJECTIVES: The objective of this review was to assess the effect of androgen treatment of men among couples where failure to conceive has been attributed to idiopathic oligo- and/or asthenospermia. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched". SELECTION CRITERIA: Randomised trials of mesterolone or testosterone undecanoate versus placebo or no treatment (stimulatory therapy), or testosterone enanthate or testosterone undecanoate versus placebo or no treatment (rebound therapy) in couples where subfertility is attributed to male factor. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. MAIN RESULTS: Eleven trials involving 930 patients were included. For stimulatory therapy, androgens had little effect on endocrinal outcomes and sperm parameters. The rate of pregnancy after androgens with stimulatory effect compared to no treatment or placebo was also similar (odds ratio 1.10, 95% confidence interval 0.75 to 1.61). In rebound therapy, no difference was found in sperm parameters. The pregnancy rate after androgens with rebound effect also showed no difference compared to no treatment or placebo (odds ratio 1.60, 95% confidence interval 0.42 to 6.16). Adverse effects such as headaches and exanthema were reported. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of androgens for male subfertility. [This abstract has been prepared centrally.] PMID- 10796497 TI - Clomiphene or tamoxifen for idiopathic oligo/asthenospermia. AB - BACKGROUND: Oligo-astheno-teratospermia (sperm of low concentration, reduced motility and increased abnormal morphology) of unknown cause is common and the need for treatment is felt by patients and doctors alike. As a result, a variety of empirical, non-specific treatments have been used in an attempt to improve semen characteristics and fertility. The administration of anti-oestrogens is a common treatment because anti oestrogens interfere with the normal negative feedback of sex steroids at hypothalamic and pituitary levels in order to increase endogenous gonadotropin-releasing hormone secretion from the hypothalamus and FSH and LH secretion directly from the pituitary. In turn, FSH and LH stimulate Leydig cells in the testes, and this has been claimed to lead to increased local testosterone production, thereby boosting spermatogenesis with a possible improvement in fertility. There may also be a direct effect of anti oestrogens on testicular spermatogenesis or steroidogenesis. This review considers the available evidence of the effect of both Clomiphene citrate and tamoxifen, both of which have a predominant anti-oestrogenic effect, for idiopathic oligo and/or asthenospermia. OBJECTIVES: The objective was to assess the effects of treating subfertile men with anti-oestrogens (clomiphene or tamoxifen) on pregnancy rates among couples where subfertility has been attributed to idiopathic oligo- and/or asthenospermia. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched". SELECTION CRITERIA: Randomised trials of anti-oestrogen therapy for 3 months or more compared to placebo or no placebo for subfertile males among couples where subfertility is attributed to male factor. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Any differences were resolved with a third reviewer. MAIN RESULTS: Ten studies involving 738 men were included. Five of the trials did not specify method of randomisation. Anti oestrogens had a positive effect on endocrinal outcomes, such as serum testosterone levels. In trials with secure randomisation there was no difference in the pregnancy rate between the anti-oestrogen groups and the control groups (odds ratio 1.26, 95% confidence interval 0.99 to 1.56). The overall pregnancy rate for these five trials was 15.4% compared to the spontaneous rate of 12.5% in the control groups. These odds increased to 1.56 (95% confidence interval 0.99 to 2.19) when all 10 trials were included, but this result is likely to be artificially inflated. REVIEWER'S CONCLUSIONS: Anti-oestrogens appear to have a beneficial effect on endocrinal outcomes, but there is not enough evidence to evaluate the use of anti-oestrogens for increasing the fertility of males with idiopathic oligo-asthenospermia. PMID- 10796498 TI - Bromocriptine for idiopathic oligo/asthenospermia. AB - BACKGROUND: Oligo-astheno-teratospermia (sperm of low concentration, reduced motility and increased abnormal morphology)of unknown cause is common and the need for treatment is felt by patients and doctors alike. As a result, a variety of empirical, non-specific treatments have been used in an attempt to improve semen characteristics and fertility. Whilst bromocriptine treatment for reducing prolactin levels in hyperprolactinaemic males (as in females), and, in the treatment of hypogonadotropic hypogonadism with hyperprolactinaemia, is beneficial, it has also been used for oligospermic men in the absence of any endocrinopathy. Prolactin may play a direct role in spermatogenesis and hormone production. It has also been claimed that in oligospermic men with normal gonadotrophins mean prolactin levels are higher and that hyperprolactinaemia is more common compared to fertile men. It has been proposed that the administration of bromocriptine under these circumstances might counteract a prolactin-induced block on the action of gonadotrophins on the testicles and, subsequently, that the reduction in prolactin levels might lead to an improvement in semen parameters and fertility. Although it is not licensed for use in male infertility, bromocriptine has been used for normogonadotrophic individuals with oligospermia and normal or sligthly elevated prolactin levels. This review considers the available evidence of the effect of bromocriptine therapy for normoprolactinaemic males with idiopathic oligo and/or asthenospermia. OBJECTIVES: The objective of this review was to assess the effects of bromocriptine on pregnancy rates among couples where subfertility has been attributed to idiopathic oligo- and/or asthenospermia. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched". SELECTION CRITERIA: Randomised trials of oral bromocriptine versus placebo or no treatment for couples with subfertility attributed to male factor. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and any disagreements were resolved by discussion with other reviewers. MAIN RESULTS: Four studies were included. The method of randomisation was not specified in any of the trials, which were all of crossover design. Compared with placebo, bromocriptine was associated with a significant reduction in serum prolactin levels (weighted mean difference -195.3 micro international units per litre, 95% confidence interval -276.5 to -114). No effects on sperm parameters were seen. There was also no effect on pregnancy rates observed between bromocriptine and placebo (0.70 odds ratio, 95% confidence interval 0.15 to 3.24). REVIEWER'S CONCLUSIONS: Bromocriptine appears to reduce prolactin levels in subfertile men with normal gonadotrophic function. There is not enough evidence to show that bromocriptine is helpful in improving fertility. PMID- 10796499 TI - Physician advice for smoking cessation. AB - BACKGROUND: Health care professionals frequently advise patients to improve their health by stopping smoking. Such advice may be brief, or part of more intensive interventions. OBJECTIVES: The aims of this review were to assess the effectiveness of advice from physicians in promoting smoking cessation; to compare minimal interventions by physicians with more intensive interventions; to assess the effectiveness of various aids to advice in promoting smoking cessation and to determine the effect of anti-smoking advice on disease specific and all cause mortality. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register and the Cochrane Controlled Trials Register. Date of the most recent searches: October 1998. SELECTION CRITERIA: Randomised trials of smoking cessation advice from a medical practitioner in which abstinence was assessed at least six months after advice was first provided. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the setting in which advice was given, type of advice given (minimal or intensive), and whether aids to advice were used, the outcome measures, method of randomisation and completeness of followup. The main outcome measure was abstinence from smoking after at least six months follow-up. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Subjects lost to follow up were counted as smokers. Where possible, meta-analysis was performed using a fixed effects model. MAIN RESULTS: We identified thirty-one trials, conducted between 1972 and 1997, including over 26,000 smokers. In some trials, subjects were at risk of specified diseases (chest disease, diabetes, ischaemic heart disease), but most were from unselected populations. The most common setting for delivery of advice was primary care. Other settings included hospital wards and outpatient clinics, and industrial clinics. Pooled data from 16 trials of brief advice versus no advice (or usual care) revealed a small but significant increase in the odds of quitting (odds ratio 1.69, 95% confidence interval 1.45 to 1.98). This equates to an absolute difference in the cessation rate of about 2.5%. There was insufficient evidence, from indirect comparisons, to establish a significant difference in the effectiveness of physician advice according to the intensity of the intervention, the amount of follow-up provided, and whether or not various aids were used at the time of the consultation in addition to providing advice. However, direct comparison of intensive versus minimal advice showed a small advantage of intensive advice (odds ratio 1.44, 95% confidence interval 1.23 to 1.68). In one study which determined the effect of smoking advice on mortality at twenty years, there were no statistically significant differences in death rates in the group receiving advice. REVIEWER'S CONCLUSIONS: Simple advice has a small effect on cessation rates. Additional manoeuvres appear to have only a small effect, though more intensive interventions are marginally more effective than minimal interventions. PMID- 10796500 TI - Prevention versus treatment for malaria in pregnant women. AB - OBJECTIVES: Malaria contributes to antenatal anaemia and slowing of fetal growth, especially in first-time mothers. It is thought that these effects harm the mother and baby, and interventions to prevent or mitigate the effects of malaria are often recommended. The objective of this review was to assess the effects of anti-malarial interventions in pregnant women living in malarial areas on the mother and the infant. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase. We contacted researchers in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials in pregnant women of interventions that aim to mitigate the effects of malaria in pregnancy, including drugs given routinely and mosquito control measures. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Data extraction was done by two reviewers using standard criteria. MAIN RESULTS: Fifteen trials were included. Drugs given regularly and routinely were associated with fewer episodes of fever in the mother, fewer women with severe anaemia antenatally, and higher average birthweight in infants. These effects appear to be greater in primigravidae. No difference in perinatal, neonatal and infant mortality were detected in studies of prophylaxis in all parity groups, or studies confined to women of low parity. REVIEWER'S CONCLUSIONS: Drugs locally effective for malaria when given routinely for malaria during pregnancy may reduce the incidence of low birth weight and anaemia. This effect appears to be limited to low parity women. Given the costs and inputs required to effectively deliver widescale prophylaxis programmes, we believe a large simple placebo- controlled trial testing the impact of drugs given routinely on pregnancy outcome and neonatal/infant survival is warranted. PMID- 10796501 TI - Drugs for preventing tuberculosis in HIV infected persons. AB - BACKGROUND: People with HIV have a increased risk of developing tuberculosis. Preventive therapy may help prevent progression of tuberculosis infection to disease. OBJECTIVES: The objective of this review was to assess the effects of preventive therapy with anti-tuberculosis drugs in people with HIV infection. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles were searched. Researchers in the field were contacted. SELECTION CRITERIA: Randomised trials of anti-tuberculosis drugs in people with HIV infection but without evidence of active tuberculosis. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and trial quality. Study authors were contacted for additional information. MAIN RESULTS: Six trials were included. Compared to placebo, preventive therapy was associated with a lower incidence of active tuberculosis (relative risk 0.54, 95% confidence interval 0.39 to 0.76). Risk of death (relative risk 0.96, 95% confidence interval 0.82 to 1.13) was not significantly different in the two groups. Incidence of tuberculosis was reduced in people with a positive tuberculin skin test (relative risk 0.24, 95% confidence interval 0.14 to 0.40), but was not significantly lower in those with a negative skin test (relative risk 0.87, 95% confidence interval 0.56 to 1.36). Similarly death was less frequent in those with a positive skin test who received preventive therapy (relative risk 0.77, 95% confidence interval 0.58 to 1.03), but this difference was not observed among those with a negative skin test (relative risk 1.07, 95% confidence interval 0.88 to 1.30). Each regimen (isoniazid alone, isoniazid plus rifampicin, isoniazid plus rifampicin plus pyrazinamide, rifampicin plus pyrazinamide) had similar protective effects against active tuberculosis for people with positive skin tests. REVIEWER'S CONCLUSIONS: Preventive therapy appears to be effective in reducing incidence of tuberculosis, and death from tuberculosis in HIV infected adults with a positive tuberculin skin test, at least in the short to medium term. PMID- 10796502 TI - Printed educational materials: effects on professional practice and health care outcomes. AB - BACKGROUND: It is often assumed that merely providing information in an accessible form will influence practice. Although such a strategy is still widely used in an attempt to change behaviour, there is a growing awareness that simply providing information may not lead to appropriate changes in the practice of health care professionals. OBJECTIVES: To assess the effects of printed educational materials in improving the behaviour of health care professionals and patient outcomes. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, reference lists of articles, and contacted content area experts. SELECTION CRITERIA: Randomised trials, interrupted time series analyses and non equivalent group designs with pre-post measures of interventions comparing 1. Printed educational materials versus a non intervention control; and 2. Printed educational materials plus additional implementation strategies versus printed educational materials alone. The participants were any health care professionals provided with printed educational materials aimed at improving their practice and/or patient outcomes. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eleven studies were included involving more than 1848 physicians. It proved impractical to examine the impact of interventions quantitatively because of poor reporting of results and inappropriate primary analyses. Nine studies examined comparison 1. Estimates of the benefit from printed educational materials ranged from -3% to 243.4% for provider outcomes, and from -16.1% to 175.6% for patient outcomes, although the practical importance of these changes is, at best, small. Six studies (seven comparisons) examined comparison 2. Benefits attributable to additional interventions ranged from 11.8% to 92.7% for professional behaviour, and -24.4% to 74.5% for patient outcomes. Two of the 14 estimates of professional behaviour, and two of the 11 estimates of patient outcomes were statistically significant. REVIEWER'S CONCLUSIONS: The effects of printed educational materials compared with no active intervention appear small and of uncertain clinical significance. These conclusions should be viewed as tentative due to the poor reporting of results and inappropriate primary analyses. The additional impact of more active interventions produced mixed results. Audit and feedback and conferences/workshops did not appear to produce substantial changes in practice; the effects in the evaluations of educational outreach visits and opinion leaders were larger and likely to be of practical importance. None of the studies included full economic analyses, and thus it is unclear to what extent the effects of any of the interventions may be worth the costs involved. PMID- 10796503 TI - Ribavirin for respiratory syncytial virus infection of the lower respiratory tract. AB - OBJECTIVES: Respiratory syncytial virus (RSV) is a common cause of pneumonia in infants. Ribavirin is the only antiviral therapy available against RSV. The objective of this review was to assess the effects of aerosolized ribavirin for infants with RSV lower respiratory tract infection. SEARCH STRATEGY: We searched MEDLINE from 1975 to 1999, we scanned reference lists of articles, and we contacted experts in the field. SELECTION CRITERIA: Randomized trials comparing ribavirin with placebo in infants and children with RSV infection and lower respiratory tract infection. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. Unpublished data were requested from authors when necessary. MAIN RESULTS: Ten trials were included. All trials enrolled infants below the age of six months. In four trials with 158 patients, mortality with ribavirin was 5.8% compared with 9.7% with placebo (odds ratio 0.58, 95% confidence interval 0.18 to 1.85). In three trials with 116 patients the probability of respiratory deterioration with ribavirin was 7.1% compared with 18.3% with placebo (odds ratio 0.37, 95% confidence interval 0.12 to 1.18). In three studies with 104 patients the weighted mean difference in days of hospitalization was 1.9 fewer days with ribavirin (95% confidence interval +0.9 to -4.6) and the difference in days of ventilation was 1.2 fewer days with ribavirin (95% confidence interval -0.2 to -3.4). REVIEWER'S CONCLUSIONS: Trials of ribavirin for RSV lack sufficient power to provide reliable estimates of the effects. The cumulative results of three small trials show that ribavirin reduces length of mechanical ventilator support and may reduce days of hospitalization. A large randomized controlled trial of ribavirin for ventilated and other high-risk patients is indicated. PMID- 10796504 TI - Steroid sex hormones for lower limb atherosclerosis. AB - BACKGROUND: There is accumulating evidence that steroid sex hormones have a beneficial effect on a number of risk factors for peripheral arterial disease. OBJECTIVES: The objective of this review was to determine whether exogenous steroid sex hormones are an effective treatment for patients with lower limb atherosclerosis. SEARCH STRATEGY: The Cochrane Peripheral Vascular Diseases Group trials register was searched, together with reference lists from relevant articles and reviews obtained through searches of Embase and Medline. SELECTION CRITERIA: Randomised controlled trials of steroid sex hormones in patients with lower limb atherosclerosis were selected. DATA COLLECTION AND ANALYSIS: Both reviewers extracted data and assessed trial quality independently. Whenever possible investigators were contacted to obtain information needed for the review that could not be found in published reports. MAIN RESULTS: Four trials appeared to meet the inclusion criteria, but one was excluded because of poor methodology and another is awaiting translation into English. The two remaining trials compared testosterone treatment with placebo in a total of 83 men with intermittent claudication. No trials were available which investigated the potentially beneficial effects of oestrogenic hormones in women with lower limb atherosclerosis. Testosterone therapy produced no improvement in tests of walking distance or in a variety of other objective tests for peripheral arterial disease, including venous filling time, muscle blood flow and plethysmography. The odds ratio for subjective improvement in symptoms using the combined trial results was also non-significant (odds ratio 1.28, 95% confidence interval 0.61 to 2.65). REVIEWER'S CONCLUSIONS: There is no evidence to date that short-term testosterone treatment is beneficial in male subjects with lower limb atherosclerosis. However, this might reflect limited data rather than the lack of a real effect. PMID- 10796505 TI - Silver acetate for smoking cessation. AB - BACKGROUND: Silver acetate produces an unpleasant taste when combined with cigarettes, thereby producing an aversive stimulus. It has been marketed in various forms with the aim of extinguishing the urge to smoke, by pairing the urge with an unpleasant stimulus. OBJECTIVES: The aim of this review was to determine the effectiveness of silver acetate products (gum, lozenge, spray) in promoting smoking cessation. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: We included randomised trials of silver acetate for smoking cessation with reports of smoking status at least six months after the beginning of treatment. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the dose and form of silver acetate, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was biochemically validated abstinence from smoking after at least six months follow-up in patients smoking at baseline. Subjects lost to follow-up were counted as continuing smokers. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: Two studies provided long-term follow-up data on patients randomised to silver acetate or placebo. In one of these studies, there was a third arm, randomised to 2mg nicotine gum. The combined odds ratio for quitting for silver acetate vs placebo was 1.05 (95% confidence interval 0.63 to 1.73). REVIEWER'S CONCLUSIONS: Existing trials show little evidence for a specific effect of silver acetate in promoting smoking cessation. The confidence intervals for the ratio are quite wide. However, the upper limit of the confidence intervals for a positive effect equates to an absolute increase in the smoking cessation rate of about 4%. Any effect of this agent is therefore likely to be smaller than nicotine replacement therapy. The lack of effect of silver acetate may reflect poor compliance with a treatment whose rationale is to create an unpleasant stimulus. PMID- 10796506 TI - Corticosteroids for preventing relapse following acute exacerbations of asthma. AB - BACKGROUND: Acute asthma is responsible for many emergency department visits annually. Between 12-16% will relapse to require additional interventions within two weeks of ED discharge. Treatment of acute asthma is based on rapid reversal of bronchospasm and reducing airway inflammation and this review examines the evidence for using systemic corticosteroids to improve outcomes after discharge from the ED. OBJECTIVES: To determine the benefit of corticosteroids (oral, intramuscular, or intravenous) for the treatment of asthmatic patients discharged from an acute care setting (i.e. usually the emergency department) after assessment and treatment of an acute asthmatic exacerbation. SEARCH STRATEGY: The Cochrane Airways Group "Asthma and Wheez* RCT" register was searched using the terms: a) Asthma OR Wheez* b) Glucocorticoid OR Steroid* AND c) Exacerbat* OR Relapse* OR Emerg*. In addition, authors of all included studies were contacted to determine if unpublished studies which met the inclusion criteria were available. Bibliographies from included studies, known reviews and texts were also searched for additional citations. SELECTION CRITERIA: Only randomized controlled trials were eligible for selection. Studies were included in this review if they dealt with the outpatient treatment of asthmatic exacerbations using glucocorticoids at discharge and reported either relapse rate or PFTs. Two independent reviewers first identified potentially relevant studies and then selected articles for inclusion. Methodological quality was assessed independently by two reviewers. Agreement was assessed using kappa (k) statistics. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers; authors were contacted to verify the extracted data and clarify missing information. When author contact was unsuccessful, missing data were estimated from graphs where possible. Sensitivity, sub-group and overall analyses were performed using the Cochrane Review Manager. MAIN RESULTS: A search that yielded 229 references identified 169 (73%) original publications. Reviewers identified 8 studies for potential inclusion (k =0.76); 18 references were added by searching publication reference lists and contact with authors. Of these 26 articles, a total of 7 were included in the overview. Two studies used intramuscular corticosteroids, five studies used oral corticosteroids. Significantly fewer patients in the corticosteroid group relapsed to receive additional care in the first week (odds ratio (OR) 0.35; 95% confidence interval (CI): 0.17, 0.73). This favourable effect was maintained over the first 21 days (OR 0.33; 95% CI: 0.13, 0.82). Patients receiving corticosteroids had less need for beta-agonists (weighted mean difference (WMD) -3.3 activations/day; 95% CI: 5.5, -1.0). Changes in pulmonary function tests (SMD 0.045; 95% CI: -0.47, 0.56) and side effects (SMD 0.03; 95% CI : -0.38, 0.44) in the first 7-10 days, while rarely reported, showed no differences between the treatment groups. Statistically significant heterogeneity was identified for the side effect results; all other outcomes were homogeneous. It appears that IM corticosteroids are similarly efficacious to a 7-10 day tapering course of oral agents. From these results, as few as 13 patients need to be treated to prevent relapse to additional care after an exacerbation of asthma. REVIEWER'S CONCLUSIONS: A short course of corticosteroids following assessment for an acute exacerbation of asthma significantly reduces the number of relapses to additional care and decreases beta-agonist use without an apparent increase in side effects. Intramuscular corticosteroids appear as effective as oral agents. PMID- 10796507 TI - Tacrine for Alzheimer's disease. AB - BACKGROUND: Tacrine is one of the first drugs to be widely marketed for the loss of memory and intellectual decline in Alzheimer's disease. The alleged success of tacrine in the treatment of these symptoms has been heralded as confirmation of the cholinergic theory of Alzheimer's disease. However, the efficacy of tacrine for symptoms of dementia remains controversial. This is reflected by the low rate of prescription of tacrine in countries where it is approved and the lack of approval by several regulatory authorities in Europe and elsewhere. The uncertainty about the efficacy of tacrine is due to the difficulties in interpretation of the results from the clinical trials. The reasons for this are the small effects of tacrine compared to placebo for all outcomes; the high incidence of adverse events; the lack of benefit observed in several trials; the use of cross-over designs and their associated methodological problems in a disease like dementia; the use of different measurement scales to assess outcome in different trials; and the problem of high dropout rates. OBJECTIVES: To determine the clinical efficacy of tacrine for the symptoms of Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia Group Register of Clinical Trials was searched using the terms 'tacrine', 'tetrahydroaminoacridine' and 'THA' (see the Group's search strategy for full details). SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with tacrine was administered for more than a day and compared to placebo in patients with dementia of the Alzheimer's type. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers, pooled if appropriate and possible, and the pooled odds ratios (95%CI) or the average differences (95%CI) were estimated. Where possible, intention-to-treat data were used. MAIN RESULTS: This review produced no clear results. The results were compatible with tacrine producing improvement, no change or even harm for those with Alzheimer's disease. It was not possible to use many of the published results in a combined analysis. For measures of overall clinical improvement, the intention-to-treat analyses failed to detect any difference between tacrine and placebo (OR 0.87; 95%CI 0.61 - 1.23). Behavioural disturbance, as measured by the Alzheimer's Disease Assessment Scale-noncognitive, failed to detect any difference between tacrine and placebo (SMD -0.04; 95%CI -0.52 - 0.43). For cognition function, the effect of tacrine was not statistically significantly different from placebo for the MiniMental State Examination score (0-30; high =good) (SMD 0.14; 95%CI -0.02 - 0.30) and was barely statistically significantly in favour of treatment for the Alzheimer's Disease Assessment Scale-cognitive scale (SMD -0.22; 95%CI -0.32 - -0.13). Adverse events were not reported in a systematic way in the different trials, making formal comparison difficult. Raised serum liver enzymes was the major reason for withdrawal. The odds ratio for withdrawal due to an adverse event was significantly different from one, the control group experienced fewer events (OR 5.7; 95%CI 4.1-7.9). Gastrointestinal side effects (diarrhoea, anorexia, dyspepsia and abdominal pain) were the other major cause of adverse events and for withdrawal, and the odds ratio for withdrawal was also significantly different from one in favour of the control group (OR 3.8; 95%CI 2.8-5.1). No deaths were reported in any of the studies during the trial period, up to six months. REVIEWER'S CONCLUSIONS: This review provides no convincing evidence that tacrine is a useful treatment for the symptoms of Alzheimer's disease. However, as so few trials presented data in a format suitable for pooling, the results of this review may be modified when further data from all relevant trials are included. There is an urgent need for the independent evaluation of the data already existing in the trials but not accessible through published or grouped data. A PMID- 10796508 TI - Vitamin E for neuroleptic-induced tardive dyskinesia. AB - BACKGROUND: Neuroleptic (antipsychotic) medication is used extensively to treat people with chronic mental illnesses. However, it is associated with a wide range of adverse effects, including movement disorders such as tardive dyskinesia (TD). Vitamin E has been proposed as a treatment to prevent or decrease the severity of TD. OBJECTIVES: To determine the clinical effects of vitamin E for people with schizophrenia or other chronic mental illnesses who also developed neuroleptic induced tardive dyskinesia. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1998), The Cochrane Schizophrenia Group's Register (September 1998), EMBASE (1980-98), LILACS (1982-96), MEDLINE (1966-98), PsycLIT (1974-98), SCISEARCH, handsearching the references of all identified studies and contacting the first author of each included trial. SELECTION CRITERIA: Reports identified in the search were included if they were controlled trials dealing with people with neuroleptic-induced TD and schizophrenia or other chronic mental illness who had been randomly allocated to either vitamin E or to a placebo or no intervention. DATA COLLECTION AND ANALYSIS: Data were independently extracted from these trials by each reviewer and Peto odds ratios (OR) or average differences, with the 95% confidence intervals (CI) were estimated. The reviewers assumed that people who dropped out had no improvement. MAIN RESULTS: Eight studies were included, and another three are currently awaiting further data from authors. The overall results for both, 'clinically relevant improvement' and 'any improvement' of TD symptoms, were in favour of vitamin E (OR 0.16, CI 0.04-0.7, NNT 5 CI 2.-32 and OR 0.23, CI 0.10-0.55, NNT 4 CI 2. 5-12 respectively). People who had not used vitamin E showed more deterioration of their symptoms (OR 0.20, CI 0.04-0.93). No difference could be found regarding the presence of adverse effects or leaving the study early before the end of study. There is no trial based information regarding the effect of vitamin E for those with early onset of TD. REVIEWER'S CONCLUSIONS: Small trials with uncertain quality of randomisation, tend to suggest that vitamin E improves the symptoms of TD. Methodological problems such as small sample size, short term interventions, and inappropriate use of crossover design need to be dealt with in any future studies. The results of a recently completed trial involving 158 participants are eagerly awaited. PMID- 10796510 TI - Drugs for treating neurocysticercosis (tapeworm infection of the brain). AB - BACKGROUND: Anthelminthic drugs may shrink brain cysts in neurocysticercosis, but can also cause severe adverse effects. OBJECTIVES: The objective of this review was to assess the effects of drug treatment in human neurocysticercosis in relation to survival, cyst persistence, subsequent seizures and hydrocephalus. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register and Medline. We contacted researchers and experts in the field and drug manufacturers. SELECTION CRITERIA: Randomised or quasi-randomised trials comparing a cysticidal drug with a placebo or a control group receiving symptomatic therapy, in patients with neurocystercosis. DATA COLLECTION AND ANALYSIS: Assessment of trial quality and data extraction was done independently by two reviewers. MAIN RESULTS: Four studies involving 305 people met the inclusion criteria. None reported on withdrawal of anticonvulsant therapy, headache relief, disability or death as outcomes. A difference just approaching significance was detected between cysticidal therapy and placebo in relation to cyst persistence up to six months (relative risk 0.83, 95% confidence interval 0.70 to 0.99). Two trials reported on seizures after one to two years follow-up and found no difference (relative risk 0.95, 95% 0.59 to 1.51). There was no difference detected for hydrocephalus (relative risk 2.19, 95% confidence interval 0.29 to 16.55). REVIEWER'S CONCLUSIONS: There is insufficient evidence to assess whether cysticidal therapy in neurocysticerosis is associated with beneficial effects. PMID- 10796511 TI - Drugs for treating giardiasis. AB - BACKGROUND: There can be a high rate of recurrence of disease after initial drug treatment for giardiasis. These drugs also have a range of adverse effects. OBJECTIVES: The objective of this review was to assess the effects of drug treatments for giardiasis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline and Embase, Current Contents, reference lists of articles. SELECTION CRITERIA: Randomised and quasi-randomised trials of drug therapy for giardiasis compared with placebo or another drug. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Thirty four trials were included. Only one trial was without serious methodological flaws. Compared with placebo, drug treatment was associated with an improved cure rate (odds ratio 11.5, 95% confidence interval 2.3 to 58). Metronidazole treatment longer than three days had a better parasitological cure rate than other long treatment courses (odds ratio 2.4, 95% confidence interval 1.3 to 4.4), but there was significant heterogeneity between the trials. Single dose therapy appeared equally effective as longer treatment courses (odds ratio 0.33, 95% confidence interval 0.08 to 1.34). Within the single dose regimens, tinidazole had a comparable parasitological cure rate to other short therapies (odds ratio 3.4, 95% confidence interval 0.95 to 12), but had a higher clinical cure rate (odds ratio 5.3, 95% 2.7-10.7). REVIEWER'S CONCLUSIONS: A single dose of tinidazole appears to give the highest clinical cure rate for giardiasis with relatively few adverse effects. PMID- 10796512 TI - Interventions for treating trichomoniasis in women. AB - OBJECTIVES: Around 120 million women worldwide suffer from Trichomonas vaginalis vaginitis every year. The infection is sexually transmitted and is believed to facilitate HIV transmission. The objective of the review is to assess the effects of various treatment strategies for trichomoniasis in women. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, and EMBASE. Trials were also identified from reference lists of reviews, through pharmaceutical companies, and by informal discovery. Only published data were used in this review. Date of the most recent search: May, 1999. SELECTION CRITERIA: Randomized or quasi-randomized trials in women with trichomoniasis of different treatment strategies, different antitrichomonal drugs or doses were eligible. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extracted by two reviewers independently using standard criteria. MAIN RESULTS: Fifty-two trials were included. Nitroimidazoles seem to be effective in achieving parasitological cure in the short term follow-ups. Partner treatment can be effective in decreasing longer term re-infection rates. REVIEWER'S CONCLUSIONS: Parasitological cure can be achieved by single oral dose of nitroimidazoles. Further research should focus on developing effective partner treatment strategies to prevent re-infections and reduce trichomoniasis prevalence. PMID- 10796513 TI - Antibiotics for acute otitis media in children. AB - BACKGROUND: Acute otitis media is one of the most common diseases in early infancy and childhood. Antibiotic use for acute otitis media varies from 31% in the Netherlands to 98% in the USA and Australia. OBJECTIVES: The objective of this review was to assess the effects of antibiotics for children with acute otitis media. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, Index Medicus, Current Contents and reference lists of articles from 1958 to January 1999. SELECTION CRITERIA: Randomised trials comparing antimicrobial drugs with placebo in children with acute otitis media. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Nine trials were eligible but only six trials, with a total of 1,962 children, included patient-relevant outcomes. The methodological quality of the included trials was generally high. All trials were from developed countries. The trials showed no reduction in pain at 24 hours, but a 34% relative reduction (95% confidence interval 16% to 48%) in pain at two to seven days. Since approximately 85% of patients will have settled spontaneously in this time, this means an absolute reduction of about 5% or that about 20 children must be treated with antibiotics to prevent one child having some pain after two days. There was no effect of antibiotics on deafness, as measured by subsequent tympanometry, other complications, or recurrence. However, audiometry was done in only two studies and incompletely reported, and there were few serious complications seen in these trials: only one case of mastoiditis occurred (in a penicillin treated group). One semi-randomised trial in Sweden in 1954 reported a rate of 17% in the untreated group versus none in the penicillin treated groups. REVIEWER'S CONCLUSIONS: Antibiotics provide a small benefit for acute otitis media in children. As most cases will resolve spontaneously, this benefit must be weighed against the possible adverse reactions. Antibiotic treatment may play an important role in reducing the risk of mastoiditis in populations where it is more common. PMID- 10796514 TI - Doxapram for ventilatory failure due to exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: COPD is a progressive illness and in the later stages, exacerbations may lead to ventilatory failure. The combination of hypoxia and hypercapnia can lead to coma and death. Correction of these blood gas abnormalities is a medical emergency. Doxapram is a respiratory stimulant used to stimulate breathing in this setting. OBJECTIVES: The objective of this review was to assess the effects of doxapram on gas exchange and clinical outcomes in people with ventilatory failure due to acute exacerbations of chronic obstructive pulmonary disease. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. We also contacted experts in the field, study authors and drug companies. SELECTION CRITERIA: Randomised trials comparing doxapram with other treatments or placebo in people with ventilatory failure due to exacerbations of chronic obstructive pulmonary disease. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Three trials involving 127 people were included. The trials were of variable quality. Doxapram was marginally superior to placebo in preventing blood gas deterioration (odds ratio 0.38, 95% confidence interval 0.14 to 1.02). In one small study, of 17 patients, doxapram and non-invasive ventilation appeared equally effective in terms of blood gases changes, although there were slightly more deaths with doxapram (odds ratio 11.34, 95% confidence interval 1.00 to 128.03). REVIEWER'S CONCLUSIONS: Doxapram may improve blood gas exchange in the short term, but newer techniques such as non-invasive ventilation may be more effective. PMID- 10796515 TI - Antibiotics for acute maxillary sinusitis. AB - OBJECTIVES: For adults seeking care in ambulatory practices, sinusitis is the most common diagnosis treated with antibiotics. We examined whether antibiotics are indicated for acute sinusitis, and if so, which antibiotic classes are most effective. SEARCH STRATEGY: Relevant studies were identified from searches of MEDLINE and EMBASE in October 1998, contacts with pharmaceutical companies and bibliographies of included studies. SELECTION CRITERIA: Randomized trials were eligible that compared antibiotic to control or antibiotics from different classes for acute maxillary sinusitis. Additional criteria were diagnostic confirmation by radiograph or sinus aspiration, outcomes that included clinical cure or improvement and a sample size of 30 or more adults. Of 1784 potentially relevant studies, two or more reviewers identified 32 studies meeting selection criteria. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by 2 persons and synthesized descriptively. Some data were analyzed quantitatively using a random effects model. Primary outcomes were a) clinical cure and b) clinical cure or improvement. Secondary outcomes were radiographic improvement, relapse rates, and dropouts due to adverse effects. MAIN RESULTS: Thirty-two trials, involving 7,330 subjects evaluated antibiotic treatment for acute maxillary sinusitis. Major comparisons were antibiotic vs. control (n=5); newer, non-penicillin antibiotic vs. penicillin class (n=10); and amoxicillin clavulanate vs. other extended spectrum antibiotics (n=10). Most trials were conducted in otolaryngology settings. Only 5 trials described adequate allocation and concealment procedures; 10 were double-blind. Compared to control, penicillin improved clinical cures [relative risk (RR) 1.72, 95% CI 1.00 to 2.96]. Treatment with amoxicillin did not significantly improve cure rates (RR 2.06; 95% CI 0.65 to 6.53), but there was significant variability between studies. Radiographic outcomes were improved by antibiotic treatment. Comparisons between classes of antibiotics showed no significant differences: newer non-penicillins vs. penicillins (RR for cure 1.07; 95% CI 0.99 to 1.17); newer non-penicillins vs. amoxicillin-clavulanate (RR for cure 1.01, 95% CI 0.97 to 1.04). Compared to amoxicillin-clavulanate, dropouts due to adverse effects were significantly lower for cephalosporin antibiotics. Relapse rates within one month of successful therapy were 5%. REVIEWER'S CONCLUSIONS: For acute maxillary sinusitis confirmed radiographically or by aspiration, current evidence is limited but supports penicillin or amoxicillin for 7 to 14 days. Clinicians should weigh the moderate benefits of antibiotic treatment against the potential for adverse effects. PMID- 10796516 TI - Antibiotics for acute bronchitis. AB - BACKGROUND: Antibiotic treatment of acute bronchitis, which is one of the most common illnesses seen in primary care, is controversial. Most clinicians prescribe antibiotics in spite of expert recommendations against this practice. OBJECTIVES: People with acute bronchitis may show little evidence of bacterial infection. If effective, antibiotics could shorten the course of the disease. However if they are not effective, the risk of antibiotic resistance may be increased. The objective of this review was to assess the effects of antibiotic treatment for patients with a clinical diagnosis of acute bronchitis. SEARCH STRATEGY: We searched Medline, Embase, reference lists of articles and the authors' personal collections up to 1996, and Scisearch from 1989 to 1996. SELECTION CRITERIA: Randomised trials comparing any antibiotic therapy with placebo in acute bronchitis. DATA COLLECTION AND ANALYSIS: At least two reviewers extracted data and assessed trial quality. MAIN RESULTS: Eight trials involving 750 patients aged eight to over 65 and including smokers and non-smokers were included. The quality of the trials was variable. A variety of outcome measures were assessed. In many cases, only outcomes that showed a statistically significant difference between groups were reported. Overall, patients receiving antibiotics had slightly better outcomes than did those receiving placebo. They were less likely to report feeling unwell at a follow up visit (odds ratio 0.42, 95% confidence interval 0.22 to 0.82), to show no improvement on physician assessment (odds ratio 0.43; 0.23 to 0.79), or to have abnormal lung findings (odds ratio 0.33, 95% confidence interval 0.13 to 0.86), and had a more rapid return to work or usual activities (weighted mean difference 0.7 days earlier, 95% confidence interval 0.2 to 1. 3). Antibiotic-treated patients reported significantly more adverse effects (odds ratio 1.64; 1.05 to 2.57) such as nausea, vomiting, headache, skin rash or vaginitis. REVIEWER'S CONCLUSIONS: Antibiotics appear to have a modest beneficial effect in the treatment of acute bronchitis, with a corresponding small risk of adverse effects. The benefits of antibiotics may be overestimated in this analysis because of the tendency of published reports to include complete data on only the outcomes found to be statistically significant. PMID- 10796517 TI - Antibiotics for the common cold. AB - BACKGROUND: The common cold is caused by viruses which cannot be helped by antibiotics. OBJECTIVES: The objective of this review was to assess the effects of antibiotics for the common cold. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, EMBASE, the Family Medicine Database, and reference lists of articles, and we contacted principal investigators. The most recent search was in December 1998. SELECTION CRITERIA: Randomised trials comparing any antibiotic therapy with placebo in acute upper respiratory tract infections. DATA COLLECTION AND ANALYSIS: Both reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Main results: Seven trials involving 2056 people aged between six months and 49 years were included. The overall quality of the included trials was variable. People receiving antibiotics did not do better in terms of cure or improvement than those on placebo (odds ratio 0.95, 95% confidence interval 0.70 to 1.28 fixed effects model). One study found a significant benefit for antibiotics compared with placebo for runny nose (clear or purulent). The only other study to evaluate purulent nasal discharge found no significant benefit for antibiotics. Only one study reported work time lost with 22% of those on antibiotic treatment and 25% of those on placebo but this was not significant. Patients treated with antibiotics had a significant increase in side effects (odds ratio 2.72, 95% confidence interval 1.02 to 7.27, random effects model). REVIEWER'S CONCLUSIONS: REVIEWERS' CONCLUSIONS: There is not enough evidence of important benefits from the treatment of upper respiratory tract infections with antibiotics and there is a significant increase in adverse effects associated with antibiotic use. PMID- 10796518 TI - Antifibrinolytics for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is an important cause of ill health in women. Medical therapy, with the avoidance of possibly unnecessary surgery, is an attractive treatment option. A wide variety of medications are available to reduce heavy menstrual bleeding but there is considerable variation in practice and uncertainty about the most appropriate therapy. Plasminogen activators are a group of enzymes that cause fibrinolysis (the dissolution of clots). An increase in the levels of plasminogen activators has been found in the endometrium of women with heavy menstrual bleeding compared to those with normal menstrual loss. Plasminogen activator inhibitors (antifibrinolytic agents) have therefore been promoted as a treatment for heavy menstrual bleeding. There has been a reluctance to prescribe tranexamic acid due to possible side effects of the drugs such as an increased risk of thrombogenic disease (deep venous thrombosis). Long term studies in Sweden, however, have shown that the rate of incidence of thrombosis in women treated with tranexamic acid is comparable with the spontaneous frequency of thrombosis in women. OBJECTIVES: To determine the effectiveness of antifibrinolytics in achieving a reduction in heavy menstrual bleeding. SEARCH STRATEGY: All studies which might describe randomised controlled trials of antifibrinolytic therapy for the treatment of heavy menstrual bleeding were obtained by electronic searches of the MEDLINE 1966-1997, EMBASE 1980-1997 and the Cochrane Library. Companies producing antifibrinolytics and experts within the field were contacted for reference lists and information on unpublished trials. SELECTION CRITERIA: Randomised controlled trials in women of reproductive age treated with antifibrinolytic agents versus placebo, no treatment or any other medical (non-surgical) therapy for regular heavy menstrual bleeding within either the primary, family planning or specialist clinic settings. Women with post menopausal bleeding, intermenstrual bleeding, iatrogenic or pathological causes of heavy menstrual bleeding were excluded. DATA COLLECTION AND ANALYSIS: Fifteen eligible trials were assessed by three reviewers and eight of these did not meet with the inclusion criteria. Of the seven remaining trials, four of these could be included within the meta-analysis. The remaining three trials had a crossover design and despite contacting the authors and appropriate companies, we were unable to extract the results in a format suitable to include these within the meta-analysis. However the results are included within the text of the review for discussion. MAIN RESULTS: Antifibrinolytic therapy compared to placebo showed a significant reduction in mean blood loss (WMD -94.0 [-151.4, -36.5]) and significant change in mean reduction of blood loss (WMD -110.2 [-146. 5, -73.8]). This objective improvement was not mirrored by a patient perceived improvement in monthly menstrual blood loss (RR 2.5 [0.9, 7.3]) in the one study which recorded this outcome ( approximately approximately Edlund 1995 approximately approximately ). Antifibrinolytic agents were compared to only three other medical (non-surgical) therapies: mefenamic acid, norethisterone administered in the luteal phase and ethamsylate. In all instances, there was a significant reduction in mean blood loss (WMD -73.0 [-123.4, -22.6], WMD -111.0 [-178.5, 43.5] and (WMD -100 [-143.9, -56.1] respectively) and a strong, although non significant trend in favour of tranexamic acid in the participants' perception of an improvement in menstrual blood loss. There were no significant differences in the frequency of reported gastrointestinal side effects with tranexamic acid when compared to either NSAIDs (RR 0.9 [0.4, 2.1], oral luteal phase progestagens (RR 0.4 [0.1, 1.2]) or ethamsylate (RR 0.88 [0.3, 2.9]) when these treatments were used for heavy menstrual bleeding. (ABSTRACT TRUNCATED) PMID- 10796519 TI - Artemisinin derivatives for treating uncomplicated malaria. AB - BACKGROUND: Artemisinin derivatives are a relatively new group of drugs with antimalarial properties. As resistance to other antimalarial drugs continues to increase, artemisinin drugs may be useful alternatives. OBJECTIVES: The objective of this review was to assess the effects of artemisinin drugs for treating uncomplicated falciparum malaria. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase, Science Citation Index, Lilacs, African Index Medicus; conference abstracts and reference lists of relevant articles. We contacted organisations, researchers in the field and drug companies. SELECTION CRITERIA: Randomised and quasi-randomised trials of artemisinin derivatives, alone or in combination with other antimalarials, compared with standard antimalarial treatments, in adults or children with uncomplicated falciparum malaria. Only trials where treatment was given by mouth or suppository were included. Comparisons between different artemisinin derivatives and treatment regimens were also included. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed and data were extracted independently by the two reviewers. MAIN RESULTS: Forty-one trials involving over 5000 patients were included. Variation in study design and quality made synthesis of the data problematic. Allocation concealment was adequate in only two trials. Most data were from areas of multidrug resistant falciparum malaria in South East Asia. Compared with standard antimalarial treatments, artemisinin drugs showed fast parasite clearance and high cure rates at follow-up, provided the duration of treatment with artemisinin drugs was adequate. Combination with mefloquine improved sustained parasite clearance and was effective in multidrug resistant areas. When doses were adequate, the combination shortened the duration of treatment. We found no evidence that artemisinin drugs are more harmful than standard treatment drugs over a typical trial period of 28 days. REVIEWER'S CONCLUSIONS: The evidence suggests that artemisinin drugs are effective and safe for treating uncomplicated malaria. There is no evidence from randomised trials that one artemisinin derivative is better than the others. In areas where there is mefloquine resistance, combination therapy with an artemisinin derivative appears to improve sustained parasite clearance compared with either drug alone. PMID- 10796520 TI - Audit and feedback: effects on professional practice and health care outcomes. AB - BACKGROUND: Audit and feedback has been identified as having the potential to change the practice of health care professionals. OBJECTIVES: To assess the effects of audit and feedback on the practice of health professionals and patient outcomes. SEARCH STRATEGY: We searched MEDLINE up to June 1997, the Research and Development Resource Base in Continuing Medical Education, and reference lists of related systematic reviews and articles. SELECTION CRITERIA: Randomised trials of audit and feedback (defined as any summary of clinical performance of health care over a specified period of time). The participants were health care professionals responsible for patient care. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Thirty seven studies were included, involving more than 4977 physicians. The reporting of study methods was inadequate for almost all studies. In 31 out of 37 studies the randomisation process could not be determined. Information regarding data analysis was also lacking. For example, power calculations were not mentioned in 27 out of 37 studies. A variety of behaviours were targeted including the reduction of diagnostic test ordering, prescribing practices, preventive care, and the general management of a problem, for example hypertension. Twenty-eight studies measured physician performance, one study targeted patient outcomes in diabetes and the remaining eight studies measured both physician performance and patient outcomes. The relative percentage differences ranged from -16% to 152%. The clinical importance of the changes was not always clear. REVIEWER'S CONCLUSIONS: Audit and feedback can sometimes be effective in improving the practice of health care professionals, in particular prescribing and diagnostic test ordering. When it is effective, the effects appear to be small to moderate but potentially worthwhile. Those attempting to enhance professional behaviour should not rely solely on this approach. PMID- 10796521 TI - Audit and feedback versus alternative strategies: effects on professional practice and health care outcomes. AB - BACKGROUND: Audit and feedback has been identified as having the potential to change the practice of health care professionals. OBJECTIVES: To assess the effects of audit and feedback compared with other interventions in changing health professional practice and to assess whether the effectiveness of audit and feedback can be improved by modifying how it is done. SEARCH STRATEGY: We searched MEDLINE up to June 1997, the Research and Development Resource Base in Continuing Medical Education, and reference lists of related systematic reviews and articles. SELECTION CRITERIA: Randomised trials of audit and feedback (defined as any summary of clinical performance of health care over a specified period of time) compared with other interventions. The participants were health care providers responsible for patient care. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Twelve studies were included involving more than 2194 physicians. Seven trials with direct comparisons were included. The targeted behaviours were the management of low haemoglobin, the delivery of preventive care services (two studies), the management of high cholesterol, the performance of cervical smears, and the ordering of diagnostic tests (two studies). From the results of four trials, there is little evidence of a measurable effect of adding a complementary intervention such as a local consensus process to audit and feedback compared to audit and feedback alone. Two of three trials that compared audit and feedback to reminders reported that reminders were more effective in improving the delivery of some preventive services. REVIEWER'S CONCLUSIONS: It is not possible to recommend a complementary intervention to enhance the effectiveness of audit and feedback. Reminders might be more effective than audit and feedback to improve the delivery of some preventive services but the results are not striking. Few trials have investigated the effect of varying different characteristics of the audit and feedback process. Consideration should be given to testing the effects of modifying important characteristics such as the content, source, timing, recipient and format. PMID- 10796522 TI - Compression bandages and stockings for venous leg ulcers. AB - OBJECTIVES: To assess the effectiveness and cost-effectiveness of compression bandaging and stockings in the treatment of venous leg ulcers. SEARCH STRATEGY: Searches of 19 databases, hand searching of journals, conference proceedings and bibliographies. Manufacturers of compression bandages and stockings and an Advisory Panel were contacted for unpublished studies. SELECTION CRITERIA: Trials that evaluated compression bandaging or stockings, as a treatment for venous leg ulcers. There was no restriction on date or language. Ulcer healing was the primary endpoint. DATA COLLECTION AND ANALYSIS: Details of eligible studies were extracted and summarised using a data extraction sheet. Data extraction was verified by two reviewers independently. MAIN RESULTS: Twenty two trials reporting 24 comparisons were identified. Compression was more effective than no compression (4/6 trials). When multi-layered systems were compared, elastic compression was more effective than non-elastic compression (5 trials). There was no difference in healing rates between 4-layer bandaging and other high compression multi-layered systems (3 trials). There was no difference in healing rates between elastomeric multi-layered systems (4 trials). Multi-layered high compression was more effective than single layer compression (4 trials). Compression stockings were evaluated in two trials. One found a high compression stocking plus a thrombo stocking to be more effective than a short stretch bandage. The second small trial reported no difference between the compression stockings and Unna's boot. There was insufficient data to draw conclusion about the relative cost-effectiveness of different regimens. REVIEWER'S CONCLUSIONS: Compression increases ulcer healing rates compared with no compression. Multi layered systems are more effective than single-layered systems. High compression is more effective than low compression but there are no clear differences in the effectiveness of different types of high compression. PMID- 10796523 TI - Cytidinediphosphocholine (CDP choline) for cognitive and behavioural disturbances associated with chronic cerebral disorders in the elderly. AB - BACKGROUND: The prevalent use of this compound in the treatment of disorders of a cerebrovascular nature does not mean that a homogeneous and consistent application of this therapy has been applied. Dosage, method of administration, and selection criteria of patients have varied. The modalities of the studies, including length of observation, severity of disturbance, and methodology of the evaluation of the results were also heterogeneous. In spite of uncertainties about its efficacy, CDP-choline is a frequently prescribed drug for cognitive impairment in several European countries, especially when the clinical picture is predominantly one of cerebrovascular disease, hence the need for this review. OBJECTIVES: The objective is to assess the efficacy of CDP-choline (cytidinediphosphocholine) in the treatment of cognitive, emotional, and behavioural deficits associated with chronic cerebral disorders of the elderly. SEARCH STRATEGY: The CDCIG register of trials was searched for all relevant, non animal randomised controlled trials using the terms CDP-choline/CDP, Citicoline, Cytidine Diphosphate Choline and Diphosphocholine. The Psychlit (1974-1996), Psychiatry (1980-1996) and MEDLINE electronic databases have been searched independently by the reviewers. The reviewers have also contacted manufacturers of CDP-choline. SELECTION CRITERIA: All relevant, non-animal, unconfounded, double-blind, placebo-controlled, randomised trials of CDP-choline in cognitive impairment due to chronic cerebral disorders will be considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed the included studies, extracted the data, and pooled it when appropriate and possible. The pooled odd ratios (95% CI) or the average differences (95% CI) were estimated. No intention-to-treat data were available from the studies included. MAIN RESULTS: Seven of the included studies observed the subjects for a period between 20 to 30 days, one study was of 6 weeks duration, 2 studies used cycles extending over 2 and 3 months and one study observed continuous administration over 3 months. The studies were heterogeneous in dose, inclusion criteria for subjects, and outcome measures. Results are reported for the domains of attention, memory testing, behavioural rating scales, global clinical impression and tolerability. There was no significant evidence of a beneficial effect of CDP choline on attention. There were modest, but significant, beneficial effects of CDP-choline on memory function and behaviour. For the outcome of clinical global impression, the odds ratio for improvement in the subjects treated with CDP Choline as opposed to the subjects treated with placebo was 8.89 [5.19, 15.22]. The drug was well tolerated. REVIEWER'S CONCLUSIONS: There is some evidence that CDP Choline has a positive effect on memory and behaviour in at least the short term. The evidence of benefit from global impression is stronger, but is still limited by the duration of the studies. There is evidence that the effect of treatment is more homogeneous for patients with cognitive impairment secondary to cerebrovascular disorder. Other studies with a more appropriate length of treatment are recommended owing to the chronic and irreversible nature of the disorders for which this treatment is indicated. PMID- 10796524 TI - Anti-tuberculous therapy for maintaining remission of Crohn's disease. AB - OBJECTIVES: To evaluate the effects of anti-tuberculous therapy for the maintenance of remission in patients with Crohn's disease. SEARCH STRATEGY: We searched the Inflammatory Bowel Disease Trials Register, the Cochrane Controlled Trials Register and MEDLINE from 1966 to 1998 (supplemented by a manual search of Index Medicus from 1966 to 1994). We also searched for abstracts in Gut, Gastroenterology, and The American Journal of Gastroenterology from 1990 to 1996. Date of most recent search: August 1998. SELECTION CRITERIA: Randomized trials of anti-tuberculous therapy in patients with Crohn's disease. DATA COLLECTION AND ANALYSIS: Data on the number of patients maintaining remission for each treatment group were abstracted. These data were pooled to yield Mantel-Haenszel odds ratios and numbers needed to treat for maintenance of remission in treated versus control groups. MAIN RESULTS: A total of seven randomized trials which included 355 patients were identified. Two trials used anti-tuberculous therapy (clofazimine or clofazimine, rafmpin, ethambutol, and dapsone) in combination with corticosteroids to induce remission. Maintenance therapy consisted of the anti-tuberculous agents without corticosteroids. Control patients received corticosteroids to induce remission but no anti-tuberculous therapy. The analysis of all seven trials yielded an odds ratio for maintenance of remission of 1.36 (95% CI 0.87-2. 13). Removing the two studies that were published as abstracts did not significantly affect this result: the pooled odds ratio was 1.14 (95% CI 0.71-1.83). The two trials reported as abstracts were excluded from subgroup analyses because they did not include any information on adjunct therapy. Subgroup analysis of the two trials which used steroids to induce remission yielded an odds ratio for maintenance of remission of 3.37 (95% CI 1.38-8.24). The number needed to treat was three. However, these two trials included only 89 patients, and the results should be interpreted with caution. The remaining three trials compared the combination of anti-tuberculous therapy and 'standard therapy' with 'standard therapy alone'. The pooled odds ratio was 0.70 (95% CI 0.39-1.25). REVIEWER'S CONCLUSIONS: Anti-tuberculous therapy may be effective in maintaining remission in patients with Crohn's disease when remission has been induced with corticosteroids combined with anti-tuberculous therapy. However, the results which support this conclusion come from a subgroup of only two trials with small numbers of patients and should be interpreted with caution. Use of this therapy cannot be recommended on the basis of this evidence. PMID- 10796525 TI - Corticosteroids for maintaining remission of Crohn's disease. AB - OBJECTIVES: To evaluate the effectiveness and safety of conventional systemic corticosteroid therapy in maintaining clinical remission in Crohn's disease. SEARCH STRATEGY: A computer-assisted search of the on-line bibliographic database MEDLINE of studies published in English, French, Spanish, Italian and German between 1966 and May, 1998. Manual searches of the reference lists from the potentially relevant studies were performed in order to identify additional studies that may have been missed using the computer-assisted search strategy. Proceedings from major gastrointestinal meetings were also manually searched from 1985 to 1997 in order to identify unpublished studies. The Cochrane Controlled Trials Register and the Inflammatory Bowel Disease Review Group Trials Register were also searched. SELECTION CRITERIA: Randomized double-blind placebo controlled trials involving patients of any age with Crohn's disease in clinical remission as defined by a CDAI < 150 or by the presence of no symptoms or only mild symptoms at the time of entry into the trial. The experimental treatment consisted of oral conventional corticosteroid therapy (excluding budesonide, fluticasone, etc). Clinical disease relapse was used as the outcome measure of interest. DATA COLLECTION AND ANALYSIS: Eligible studies were selected by 4 reviewers and data were extracted onto standardized data extraction forms. Disagreements in eligibility or data extraction were resolved by consensus. Data were converted into individual 2x2 tables for each study. The presence of significant heterogeneity among studies was tested using the chi-square test. The 2x2 tables were synthesized into a summary test statistic using the pooled odds ratio and 95% confidence intervals as described by Cochran and Mantel and Haenszel (the 'odds ratio' in MetaView). A fixed effects model was used for the pooling of data. MAIN RESULTS: Four studies were initially judged as being eligible for inclusion. After obtaining additional information on one of the studies it was excluded because it was not double-blind. The total number of subjects included in the analysis at the time points of 6, 12 and 24 months were 142, 131 and 95 for the corticosteroid group and 161, 138 and 87 for the control group. The odds ratios for relapse on active treatment and the corresponding 95% confidence intervals were 0.71 (0.39, 1.31), 0.82 (0.47, 1.43) and 0.72 (0.38, 1.35) at 6, 12 and 24 months. The numbers needed to treat with corticosteroids to prevent one additional relapse were 24, 35, 15 respectively. REVIEWER'S CONCLUSIONS: The use of conventional systemic corticosteroids in patients with clinically quiescent Crohn's disease does not appear to reduce the risk of relapse over a 24 month period of follow-up. PMID- 10796526 TI - Dehydroepiandrosterone (DHEA) supplementation for cognition and well-being. AB - BACKGROUND: In view of the theoretical rationale for beneficial effects of DHEA and DHEAS in aging and dementia, we believe it is timely to undertake a thorough investigation of well-conducted studies in this area. This will provide a basis for confirmation of any effect of DHEA/S administration in humans, in large-scale and properly controlled trials, which would evaluate effective dosage, acceptable route and duration of administration and side effect profiles. This is especially pertinent at this time as DHEA is currently being sold in large quantities in health food stores, particularly in the USA. In some cases the recommended dose is different for men and women (50mg/day for men and 25mg/day for women) and the basis for this recommendation needs to be explored. OBJECTIVES: To establish whether administration of DHEA, or its sulphate, DHEAS, improves psychological well-being and/or improves cognitive function or reduces the rate of decline of cognitive function in older adults or in individuals with dementia. SEARCH STRATEGY: All available electronic databases, hand searched journals, personal communications and conference abstracts were searched for randomised controlled trials of DHEA in well-being and cognition. The total yield from searching was 415 and the detailed breakdown is given in the body of this review. SELECTION CRITERIA: All relevant randomised controlled trials of DHEA or DHEAS were considered for inclusion in the review. Studies where groups are matched, rather than randomised, were also considered. DATA COLLECTION AND ANALYSIS: Data for the specified outcomes were independently extracted by two reviewers (FAH & JvN) and cross-checked. Any discrepancies were discussed and resolved. Where possible and appropriate, data were pooled and the mean differences estimated. MAIN RESULTS: The published DHEA trials fall into 2 categories: 1. four German studies in which DHEA was administered for a period of two weeks or less; 2. a USA study in which DHEA was administered for three months. Well-being was assessed in both sets of studies and a significant improvement was reported in the longer duration USA study, while no effect was reported in the shorter duration studies. The USA study used an open-ended questionnaire for self-assessment of well-being and stated that 67% of men and 82% of women reported enhanced well-being on DHEA compared with placebo. There was no significant change on an analogue measure of libido. The German studies assessed mood and well-being with a number of standardised scales and reported no significant effects of DHEA on any of them. Only the German studies examined performance on cognitive tests, i.e. memory, verbal fluency, speed of processing, etc. They reported no significant benefit of DHEA. REVIEWER'S CONCLUSIONS: The data at present offer limited support for improvement in a sense of well-being following DHEA treatment. This effect was reported only in the longer-term study which used a crude measure of well-being. The data offer no support at present for an improvement in memory or other aspects of cognitive function following DHEA treatment, although cognitive function was only measured in the short-duration trials. In view of the growing public enthusiasm for DHEA supplementation, particularly in the USA, it is clear that high-quality trials need to be undertaken in older adults, in which (a) the duration of DHEA treatment is in excess of two weeks, (b) the number of participants is large enough to detect effects if they exist, and (c) the outcome measures include validated scales for assessment of mood and well-being, and objective tests of cognitive function. Recently, studies of DHEA supplementation in clinical depression and Alzheimer's Disease have been completed in the USA. As soon as the results are available these studies will be reviewed. Currently, two trials (in France and the USA) in normal elderly are in progress. PMID- 10796527 TI - Interventions for treating scabies. AB - BACKGROUND: Scabies is a common public health problem with an estimated prevalence of 300 million. Infestation can cause considerable discomfort and intense itching. Severe adverse effects have been reported for some drugs used to treat scabies. OBJECTIVES: The objective of this review is to assess the effects and toxicity of topical and systemic drug treatment for scabies. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase, military records, traditional medicine databases. We also contacted international specialist centres and drug manufacturers. SELECTION CRITERIA: Randomised controlled trials of any drug treatment for scabies. Tolerability and toxicity were sought in any study of humans taking any drug treatments for scabies. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trial quality and extracted data. MAIN RESULTS: Eleven trials were included (7 compared drug treatments, 2 compared treatment regimes, 1 compared the drug vehicle and 1 was a community intervention). Compared with placebo in one small trial, ivermectin was associated with a significant higher clinical cure rate at seven days. Permethrin appeared to be more effective than crotamiton for clinical and parasitic cure rates. Permethrin appeared to be better than gamma benzene hexachloride for clinical cure rates in two small trials but had no advantage in the largest trial (test for heterogeneity p< 0.001). Permethrin also appeared more effective in reducing itch persistance than gamma benzene hexachloride. There appeared to be no difference in clinical cure rates between crotamiton and gamma benzene hexachloride. Single trials assessed: the effectiveness of oral versus topical treatment (ivermectin versus benzyl benzoate); treatment vehicle (pork fat versus cold cream); and mass community treatment (ivermectin) but were too small to demonstrate an effect. No randomised trials of malathion were identified. Serious adverse drug reactions (including death and convulsions) have been reported in other studies of scabies drugs, most notably gamma benzene hexachoride and ivermectin. REVIEWER'S CONCLUSIONS: The evidence that permethrin is more effective than gamma benzene hexachloride is inconsistent. Permethrin appears to have less potential serious drugs reactions than gamma benzene hexachloride although this is not derived from trial data. More research is needed of the safety and effectiveness of ivermectin and malathion compared to permethrin, on community management, and on different regimes and vehicles for topical treatment. PMID- 10796528 TI - Endometrial resection and ablation versus hysterectomy for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) or menorrhagia is an important cause of ill health in women. Surgical treatment of HMB often follows failed or ineffective medical therapy and the definitive treatment is hysterectomy but this is a major surgical procedure with significant physical and emotional complications and social and economic costs. A number of less invasive surgical techniques (e.g. endometrial resection and laser ablation) have been developed with the purpose of removing the entire thickness of the endometrium. The benefits claimed for these therapies are reduced trauma and post-operative complications to the woman, reduced need for a general anaesthetic, direct cost savings to the health service due largely to a shift from inpatient to day case treatment and indirect cost savings to society as women return more quickly to their usual activities. However, endometrial hysteroscopic techniques are not always completely successful and additional surgical treatment is required in a proportion of cases. Although initially the resource and patient costs of these techniques are much cheaper than the cost of hysterectomy, the need for re treatment at a later stage may reduce the cost differential. Thus, the effectiveness of these techniques to improve a woman's perception of her own wellbeing long term has yet to be confirmed. OBJECTIVES: The objective of this review is to compare endometrial destruction techniques with hysterectomy by any means for the treatment of heavy menstrual bleeding (HMB). SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Sub fertility Group Register of Trials, MEDLINE, EMBASE, PsychLIT, Current Contents, Biological Abstracts, Social Sciences Index and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles and hand searching. In most cases, the first or corresponding author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of endometrial destruction techniques with hysterectomy by any means for the treatment of heavy menstrual bleeding in premenopausal women. DATA COLLECTION AND ANALYSIS: Five RCTs were identified that fulfilled the inclusion criteria for this review. For two trials, a number of publications were identified which assessed different outcomes and different follow up time points for the same patients. The reviewers extracted the data independently and odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data. Outcomes analysed included improvement in menstrual blood loss, satisfaction, change in quality of life, duration of surgery and hospital stay, time to return to work, adverse events and requirement for repeat surgery because of failure of the initial surgical treatment. MAIN RESULTS: There was a significant advantage in favour of hysterectomy in the improvement in HMB and satisfaction rates (up to 4 years post surgery) compared with endometrial destruction techniques. Although many quality of life scales reported no differences between surgery groups, there was some evidence of a greater improvement in general health for hysterectomy patients. Duration of surgery, hospital stay and recovery time were all shorter following endometrial destruction. Most adverse events, both major and minor, were significantly more likely after hysterectomy and before discharge from hospital. After discharge from hospital, the only difference that was reported for this group was a higher rate of infection. Repeat surgery because of failure of the initial treatment, either endometrial ablation or hysterectomy, was more likely after endometrial destruction than hysterectomy. (ABSTRACT TRUNCATED) PMID- 10796529 TI - Expanding the roles of outpatient pharmacists: effects on health services utilisation, costs, and patient outcomes. AB - BACKGROUND: In recent years pharmacists' roles have expanded from simply packaging and dispensing medications to working with other health care professionals and the public. OBJECTIVES: To assess the effects of expanding outpatient pharmacists' roles on health services utilisation, the costs of health services, and patient outcomes. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, MEDLINE, EMBASE, Pharmline, International Pharmaceutical Abstracts and reference lists of articles up to December 1995. We also searched the published abstracts of three meetings and hand searched five journals and two bibliographies. SELECTION CRITERIA: Randomised trials, controlled clinical trials, controlled before-and after studies and interrupted time series analyses of interventions comparing 1. Pharmacist services targeted at patients versus services delivered by other health professionals; 2. Pharmacist services targeted at patients versus no intervention; 3. Pharmacist services targeted at health professionals versus services delivered by other health professionals; and 4. Pharmacist services targeted at health professionals versus no intervention. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Fourteen studies were included involving more than 1991 patients. In the one study identified for comparison 1 the relative changes in professional outcome measures ranged from a 24% increase in clinic visits to a 16% decrease in hospital admissions. Relative differences in patient outcome measures were not statistically significant. Seven studies were identified for comparison 2. Four measured process of care and demonstrated decreases in health services utilisation from -67% for hospital admissions to -564% for total ambulatory care visits, as well as decreases in the numbers and costs of drugs compared to control patients. Five measured patient outcomes and consistently reported improvements in the targeted patient condition. In the one study identified for comparison 3 the intervention delivered by the pharmacist was less successful than that delivered by physician counsellors in decreasing inappropriate antibiotic prescribing. All six studies identified for comparison 4 demonstrated that the pharmacist intervention produced the intended effect on physician prescribing practices. These studies did not measure patient outcomes. REVIEWER'S CONCLUSIONS: The limited number of studies analysed support the expanded roles of pharmacists in patient counselling and physician education. However, doubts about the generalisability of the studies, the poorly defined nature of the interventions tested, and the lack of studies including cost assessments and patient outcome data indicate that more rigorous research is needed to document the effects of outpatient pharmacist interventions. PMID- 10796530 TI - Gonadotrophin-releasing hormone analogues for pain associated with endometriosis. AB - BACKGROUND: Endometriosis is a common gynaecological condition that frequently presents with the symptom of pain. The precise pathogenesis (mode of development) of endometriosis is unclear but it is evident that endometriosis arises by the dissemination of endometrium to ectopic sites and the subsequent establishment of deposits of ectopic endometrium. The observation that endometriosis is rarely seen in the hypo-oestrogenic (low levels of oestrogen) post-menopausal woman led to the concept of medical treatment by induction of a pseudo-menopause using Gonadotrophin Releasing Hormone Analogues (GnRHas). When administered in a non pulsatile manner (the pituitary is normally stimulated by pulses of natural GnRH and all analogues act on the pituitary at a constant level) their use results in down regulation (switching off) of the pituitary and a hypogonadotrophic hypogonadal state (low levels of female hormones due to non stimulation of the ovary). OBJECTIVES: To determine the effectiveness of Gonadotrophin Releasing Hormone analogues (GnRHas) in the treatment of the painful symptoms of endometriosis by comparing them with no treatment, placebo, other recognised medical treatments, and surgical interventions. SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility review group (please see Review Group details) was used to identify all randomised trials of the use of GnRHas for the treatment of the painful symptoms of endometriosis. SELECTION CRITERIA: Trials were included if they were randomised, and considered the effectiveness of GnRHas in the treatment of the painful symptoms of endometriosis. DATA COLLECTION AND ANALYSIS: Twenty-six studies had data appropriate for inclusion in the review. The largest group (15 studies) compared GnRHas with danazol. There are 5 studies comparing GnRHas with GnRHas plus add back therapy, 3 comparing GnRHa with GnRHa in a different form or dose, one compares them with gestrinone, one with the combined oral contraceptive pill, and one with placebo. Data was extracted independently by two reviewers. The authors of 11 studies have been contacted to clarify missing or unclear data. Only 4 have replied to date. Data on relief of pain, change in revised American Fertility Society (rAFS) scores, and side effects was collected. MAIN RESULTS: No difference was found between GnRHas and any of the other active comparators with respect to pain relief or reduction in endometriotic deposits. The side effect profiles of the different treatments were different, with danazol and gestrinone having more androgenic side effects, while GnRHas tend to produce more hypo oestrogenic symptoms. REVIEWER'S CONCLUSIONS: There is little or no difference in the effectiveness of GnRHas in comparison with other medical treatments for endometriosis. GnRHas do appear to be an effective treatment. Differences that do exist relate to side effect profiles. Side effects of GnRHas can be ameliorated by the addition of addback therapy. PMID- 10796531 TI - Guidelines in professions allied to medicine. AB - BACKGROUND: Clinical practice guidelines aim to reduce inappropriate variations in practice and to promote the delivery of evidence-based health care. OBJECTIVES: To identify and assess the effects of studies of the introduction of clinical practice guidelines in nursing (including health visiting), midwifery and other professions allied to medicine. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, MEDLINE (1975 to 1996), EMBASE, Cinahl and Sigle to 1996, the NHS Economic Evaluations Database (1994 to 1996), DHSS-Data (1983 to 1996), the Database of Abstracts of Reviews of Effectiveness (1994 to 1996) and reference lists of articles. We also hand searched the journal Quality in Health Care, made personal contact with content experts and contacted libraries identified by an expert panel. SELECTION CRITERIA: Randomised trials, controlled before-and-after studies and interrupted time series analyses of the introduction of interventions comparing 1. Clinical guidelines plus dissemination and/or implementation strategies versus no guidelines; 2. Guidelines plus dissemination and/or implementation strategies versus guidelines plus alternative dissemination and/or implementation strategies; and 3. (post hoc) Guidelines used by professions allied to medicine versus standard physician care. The participants were nurses, midwives and other professions allied to medicine. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eighteen studies were included involving more than 467 health care professionals. The reporting of study methods was inadequate for all studies. In all but one study, nurses were the targeted professional group; one study was aimed solely at dieticians. The various behaviours targeted included the management of hypertension, low back pain and hyperlipidaemia. Nine studies were identified for comparison 1. Three out of five studies observed improvements in at least some processes of care and six out of eight studies observed improvements in outcomes of care. Only one study included a formal economic evaluation, with equivocal findings. Three studies were identified for comparison 2 but it was difficult to draw firm conclusions because of poor methods. Six studies were identified for comparison 3 (post hoc). These studies generally supported the hypothesis that there was no difference between care given by nurses using clinical guidelines and standard physician care. REVIEWER'S CONCLUSIONS: There is some evidence that guideline-driven care is effective in changing the process and outcome of care provided by professions allied to medicine. However, caution is needed in generalising findings to other professions and settings. PMID- 10796532 TI - Homeopathy for chronic asthma. AB - BACKGROUND: Homeopathy involves the use, in dilution, of substances which cause symptoms in their undiluted form. It is one of the most widespread forms of complementary medicines and is also used to treat asthma. OBJECTIVES: The objective of this review was to assess the effects of homeopathy in people with chronic stable asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, the Cochrane Complementary Medicine Field trials register, the Glasgow Homeopathic Hospital database, the Muenchener Modell database and reference lists of articles. SELECTION CRITERIA: Randomised and possibly randomised trials of homeopathy for the treatment of stable chronic asthma, with observation periods of at least one week. DATA COLLECTION AND ANALYSIS: Data extraction was done by one reviewer and checked by the second reviewer. Trial quality was assessed by the reviewers. MAIN RESULTS: Three trials with a total of 154 people were included. These trials were all placebo-controlled and double blind, but of variable quality. They used three different homeopathic treatments which precluded quantitative pooling of results. The standardised treatments in these trials are unlikely to represent common homeopathic practice, where treatment tends to be individualised. In one trial, severity of symptoms was lessened in the homeopathy group compared to the placebo group. In another trial, lung function measures and medication use showed improvement in the homeopathy group compared to the placebo group. The third trial found improvement in both the homeopathy and placebo groups, but no difference between the groups. REVIEWER'S CONCLUSIONS: There is not enough evidence to reliably assess the possible role of homeopathy in asthma. As well as randomised trials, there is a need for observational data to document the different methods of homeopathic prescribing and how patients respond. PMID- 10796533 TI - Hospital-at-home versus in-patient hospital care. AB - BACKGROUND: Hospital-at-home is defined as a service that provides active treatment by health care professionals, in the patient's home, of a condition that otherwise would require acute hospital in-patient care, always for a limited period. Hospital-at-home has become a popular response to the increasing demand for acute hospital beds. OBJECTIVES: To assess the effects of hospital-at-home compared with in-patient hospital care for patients. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, the Cochrane Controlled Trials Register, MEDLINE to 1996, EMBASE to 1995, Cinahl to 1996, the Social Science Citation Index (1992 to 1995), EconLit (1969 to 1996), PsycLit (1987 to 1996), Sigle (1980 to 1995), the Medical Care supplement on economic literature (1979 to 1990), and reference lists of articles. SELECTION CRITERIA: Randomised trials of hospital-at-home care compared with acute hospital in-patient care. The participants were patients aged 18 years and over. The outcomes were mortality, clinical complications, re-admissions, cost (to the patient and family, to general practice, to the hospital and to the community), hospital days saved from the provision of hospital-at-home, discharge destination from hospital-at-home, general and disease specific health status, functional status, psychological well-being, patient satisfaction, carer satisfaction, carer burden, and staff views (including the satisfaction of doctors working in primary care). DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Five studies were included involving 866 patients. All studies were small and lacked power. No statistically significant differences were detected for patient health outcomes. Patients discharged early from hospital to hospital-at-home following elective surgery expressed greater satisfaction with care than those who remained in hospital. Carers however expressed less satisfaction with hospital-at-home compared with hospital care. Only one trial formally tested for a difference in cost, with no statistically significant difference detected for overall health care costs. REVIEWER'S CONCLUSIONS: There is insufficient evidence to assess the effects of hospital-at-home on patient outcomes or the cost to the health service. Given the heterogeneity of what hospital-at-home encompasses and the uncertainty over its effects, future research should clearly specify the type of service being provided, both at home and at hospital, and the specific patient groups. Patient health outcomes, patient and carer satisfaction, and costs should be measured, and studies should include a formal, planned economic analysis. Studies should be large enough to detect important differences and to ensure generalisability of the results. PMID- 10796534 TI - Hydergine for dementia. AB - BACKGROUND: Currently hydergine is used almost exclusively for treating patients with either dementia, or 'age-related' cognitive symptoms. Since the early eighties there have been over a dozen more clinical trials, yet hydergine's efficacy remains uncertain. Although previous reviews offer generally favorable support for hydergine's efficacy, they were, however, limited by a bias with respect to the particular clinical studies chosen (eg, the inclusion of case reports, and uncontrolled trials), and by authors' impressionistic assessments of results. Not surprisingly, there has been a lack of consensus among reviewers with regard to the efficacy of hydergine. In 1994, a meta-analysis was published by the present reviewers who reported that overall, hydergine was more effective than placebo. However they also observed that the statistical evidence for efficacy in 'possible or probable Alzheimer's disease' patients was so modest that one additional statistically non-significant trial would have reduced the results to non significance. OBJECTIVES: Because of uncertainty surrounding the efficacy of hydergine, the goals of this overview were to assess its overall effect in patients with possible dementia, and to investigate potential moderators of an effect. SEARCH STRATEGY: The Cochrane Dementia Group Register of Clinical Trials was searched using the terms 'hydergine', 'ergoloids,' 'ergoloid mesylates,' 'dihydroergocristine,' 'dihydroergocryptine,' 'dihydroergotoxine,' and 'dihydroergocornine. MEDLINE, EMBASE, and two proprietary databases were searched also. Published reviews were inspected for further sources. SELECTION CRITERIA: Trials to be included must be randomized, double-blind, parallel-group, and unconfounded comparisons of hydergine with placebo for a treatment duration of greater than 1 week in subjects with dementia or symptoms consistent with dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers, pooled where appropriate and possible, and the pooled odds ratios (95%CI) or the average differences (95%CI) were estimated. Where possible, intention-to-treat data were used. Outcomes of interest included clinical global impressions of change and comprehensive rating scales. Potential moderating variables of a treatment effect included: inpatient/outpatient status, trial duration, age, sex, medication dose, publication year, and diagnostic grouping. MAIN RESULTS: There were a total of nineteen trials that met inclusion criteria and that had data sufficient for analysis. Thirteen trials reported sufficient information to use a global rating of improvement and nine trials provided information on a comprehensive rating scale. Three trials provided both outcome measures. It was not possible to use many of the published results in a combined analysis owing to the lack of sufficient data to perform statistical analyses. For the twelve trials that used global ratings, there was a significant effect favoring hydergine (OR 3.78, 95%CI, 2.72-5.27). For the nine trials that used comprehensive ratings, there was a significant mean difference favoring hydergine (WMD 0.96, 95%CI, 0. 54-1.37). Hydergine was well tolerated in these trials, with 78% of randomized subjects available for data analyses. Greater effect sizes on global ratings were associated with younger age, and possibly higher dose, although most of the subgroup analyses were statistically insignificant. REVIEWER'S CONCLUSIONS: As in an earlier systematic review, we found hydergine to show significant treatment effects when assessed by either global ratings or comprehensive rating scales (based here on a smaller set of trials than in the earlier published systematic review because trials were required to have data that could conform with MetaView, the Cochrane Collaboration statistics software). The small number of trials available for analysis, however, limited the ability of subgroup analyses to identify statistically significant modera PMID- 10796535 TI - Insecticide-treated bednets and curtains for preventing malaria. AB - BACKGROUND: Malaria is an important cause of illness and death in many parts of the world, especially in Africa. There has been a renewed emphasis on preventive measures, both at community and at individual level. Insecticide treated bednets and curtains are a promising preventive measure. OBJECTIVES: The objective of this review was to assess the effects of insecticide-treated bednets or curtains in preventing malaria SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, Medline, Embase and the reference lists of articles. We handsearched journals, contacted researchers, funding agencies and net and insecticide manufacturers. SELECTION CRITERIA: Randomized and quasi randomized trials of insecticide-treated bednets or curtains with a sufficient dose of a suitable insecticide compared with nets without insecticide or no nets at all. Studies including pregnant women were excluded. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extracted by the reviewer. Trial inclusions/exclusions were reviewed by two independent individuals. MAIN RESULTS: Of 65 identified studies, 18 were included. Of these, 11 were randomized by cluster rather than individually. Mortality in children under five was measured in four trials. When impregnated nets were compared with plain nets or no nets, the summary relative risk was 0. 83. This translates to an estimate of protective efficacy of 17%. For treated nets compared with untreated nets, the relative risk of child mortality was 0.77. About six lives can be saved each year for every 1000 children protected with insecticide-treated nets. Insecticide-treated nets also reduced the incidence of mild malarial episodes by 48% (controls=no nets) and 34% (controls=untreated nets). REVIEWER'S CONCLUSIONS: Insecticide-treated nets appear to be effective in reducing childhood mortality and morbidity from malaria. Widespread access to insecticide-treated nets will require major financial, technical and operational inputs. PMID- 10796536 TI - Vaccines for preventing influenza in people with asthma. AB - BACKGROUND: Influenza vaccination is recommended for asthmatic patients in many countries as observational studies have shown that influenza infection can be associated with asthma exacerbations, but influenza vaccination itself has the potential to adversely affect pulmonary function. A recent overview concluded that there was no clear benefit of influenza vaccination in patients with asthma but this conclusions was not based on a systematic search of the literature. OBJECTIVES: Whilst influenza may cause asthma exacerbations, there is controversy about the use of influenza vaccinations, since they may precipitate an asthma attack in some people. The objective of this review was to assess the effects of influenza vaccination in children and adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and checked reference lists of articles. SELECTION CRITERIA: Randomised trials of influenza vaccination in children (over two years of age) and adults with asthma. Studies involving people with chronic obstructive pulmonary disease were excluded. DATA COLLECTION AND ANALYSIS: Inclusion criteria and assessment of trial quality were applied by two reviewers independently. Data extraction was done by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Nine trials were included. Four of these trials were of high quality. One further article has been included since the previous version of this review. Inclusion of the new trial has not altered the conclusions of this review. The included studies covered a wide diversity of people, settings and types of influenza vaccination, so data from the different trials were not pooled. In one trial, no protective effect of influenza vaccination against asthma exacerbation was demonstrated, but the incidence of influenza was low during the study period. A higher number of asthma exacerbations following killed influenza vaccination was found in one trial (risk difference 3 1%, 95% confidence interval 0.3% to 5.8%). When people with upper respiratory tract infections were excluded, this difference was no longer significant. A small trial using recombinant vaccine found no significant difference in asthma exacerbations between the vaccinated and placebo groups. REVIEWER'S CONCLUSIONS: There is not enough evidence to assess the benefits and risks of influenza vaccination for people with asthma. PMID- 10796537 TI - Anthelmintic drugs for treating worms in children: effects on growth and cognitive performance. AB - BACKGROUND: Helminth (worm) infections are widespread and are thought to contribute to poor nutritional status, anaemia, and impaired growth and learning in children. OBJECTIVES: To summarise the effects of anthelmintic drug treatment in children in relation to their growth and cognitive performance. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and the reference lists of articles. We also contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of drug treatment compared with placebo or no drug treatment for intestinal helminths in children. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for additional information. MAIN RESULTS: Thirty trials involving more than 1500 children were included. There was potential for bias from inadequate concealment of allocation. Studies varied in relation to target groups, drugs administered and treatment regimens. Compared to placebo or no drug treatment, drug treatment for helminths was associated with some positive effects on change in weight, height and skinfold thickness. However there was significant heterogeneity between the results of the trials. There were some positive effects on mean weight change in the trials reporting this outcome; after a single dose (any anthelminth) the pooled estimate was 0.24 kg, (95% CI 0.15 to 0.32; fixed effects model assumed); and 0.38 kg (95% CI 0.00 to 0.77; random effects model assumed). Results from trials giving multiple doses showed mean weight change under one year of follow up of 0.10 kg (95% CI 0.04 to 0.17; fixed effects assumed); or 0.15 (95% CI 0.00 to 0.30; random effects assumed). At more than one year of follow up, mean weight change was 0.12 kg (95% CI -0.02 to 0.26; fixed effects assumed) and 0.43 (95% CI -0.61 to 1.47; random effects model assumed). Results from studies of cognitive performance were mixed and inconclusive. REVIEWER'S CONCLUSIONS: There is some limited evidence that routine treatment of children in areas where helminths are common has small effects on weight gain, but this is not consistent between trials. There is insufficient evidence to know whether this intervention improves cognitive performance. Our interpretation of these results is that the current public health programme investments in this intervention, based on the expectation that there will be an improvement in growth and learning, are not based on consistent or reliable evidence. PMID- 10796538 TI - Chloroquine or amodiaquine combined with sulfadoxine-pyrimethamine for treating uncomplicated malaria. AB - BACKGROUND: Amodiaquine and chloroquine give fast relief from malaria symptoms, particularly fever. When used alone in areas where there is some parasite resistance they do not completely clear parasites from the blood in all cases, and so not all patients are cured of infection. The major disadvantage of using sulfadoxine-pyrimethamine alone is that it takes a relatively long time to relieve fever. OBJECTIVES: To assess the effectiveness of chloroquine or amodiaquine given with sulfadoxine-pyrimethamine to treat uncomplicated falciparum malaria. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, MEDLINE, EMBASE, Science Citation Index, African Index Medicus and LILACS were searched. Experts in the field and drug companies were contacted. SELECTION CRITERIA: Randomised and quasi-randomised trials of chloroquine or amodiaquine given with sulfadoxine pyrimethamine compared with either drug alone in adults or children with confirmed uncomplicated falciparum malaria. DATA COLLECTION AND ANALYSIS: Two people independently applied the inclusion criteria. Data were extracted by the reviewer and checked independently by another person. MAIN RESULTS: Five trials were included. Fever clearance time was reduced by combination therapy compared with sulfadoxine-pyrimethamine alone. Parasite clearance at day seven follow-up was not significantly different for chloroquine or amodiaquine treatment with or without sulfadoxine-pyrimethamine. Parasite clearance at day 28 was better with combination therapy compared with chloroquine or amodiaquine alone (odds ratio 14.28, 95% confidence interval 6.76 to 30.19), but not significantly better than sulfadoxine-pyrimethamine alone (odds ratio 3.17, 95% confidence interval 0.96 to 10.43). There was no evidence from the included trials of serious side effects with combination treatment. REVIEWER'S CONCLUSIONS: In areas where chloroquine or amodiaquine are still effective, despite some degree of resistance, using these drugs in combination with sulfadoxine-pyrimethamine, rather than sulfadoxine pyrimethamine alone, may make people feel better faster and improve sustained parasites clearance. PMID- 10796539 TI - Mass media interventions: effects on health services utilisation. AB - BACKGROUND: The mass media frequently cover health related topics, are the leading source of information about important health issues, and are targeted by those who aim to influence the behaviour of health professionals and patients. OBJECTIVES: To assess the effects of mass media on the utilisation of health services. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, MEDLINE, EMBASE, Eric, PsycLit, and reference lists of articles. We hand searched the journals Communication Research (February 1987 to August 1996), European Journal of Communication (1986 to 1994), Journal of Communication (winter 1986 to summer 1996), Communication Theory (February 1991 to August 1996), Critical Studies in Mass Communication (March 1984 to March 1995) and Journalism Quarterly (1986 to summer 1996). SELECTION CRITERIA: Randomised trials, controlled clinical trials, controlled before-and-after studies and interrupted time series analyses of mass media interventions. The participants were health care professionals, patients and the general public. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Seventeen studies were included. All used interrupted time series designs. Fourteen evaluated the impact of formal mass media campaigns, and three of media coverage of health related issues. The overall methodological quality was variable. Six studies did not perform any statistical analysis, and seven used inappropriate statistical tests (ie not taking into account the effect of time trend). All of the studies apart from one concluded that mass media was effective. These positive findings were confirmed by our re-analysis in seven studies. The direction of effect was consistent across studies towards the expected change. The pooled effect sizes for studies assessing the impact of mass media on similar aspects of care revealed an effect upon the utilisation of health services that could not be explained by chance alone, ranging from -1.96 (95%CI -1. 19 to -2.73) for campaigns promoting immunisation programmes, to -1. 12 (95%CI -0.49 to -2.36) for those concerning cancer screening. REVIEWER'S CONCLUSIONS: Despite the limited information about key aspects of mass media interventions and the poor quality of the available primary research, there is evidence that these channels of communication may have an important role in influencing the use of health care interventions. Those engaged in promoting better uptake of research information in clinical practice should consider mass media as one of the tools that may encourage the use of effective services and discourage those of unproven effectiveness. PMID- 10796540 TI - Methotrexate as a steroid sparing agent for asthma in adults. AB - BACKGROUND: Sustained oral corticosteroid use can lead to complications, so there is interest in identifying agents that can reduce oral steroid use in people with asthma. Methotrexate has attracted attention as a possible steroid sparing agent in patients with chronic oral steroid dependent asthma. OBJECTIVES: The objective of this review was to assess the effects of adding methotrexate to oral corticosteroids in adults with stable asthma who are dependent on oral corticosteroids. SEARCH STRATEGY: The Cochrane Airways Group trials register and reference lists of identified articles were searched. SELECTION CRITERIA: Randomised trials of the addition of methotrexate compared with placebo in adult steroid dependent asthmatics. Duration of therapy needed to be at least 12 weeks. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was carried out by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Ten trials involving a total of 185 people were included. Study design and quality, corticosteroid dosages and outcomes varied widely. There was a reduction in oral corticosteroid dose favouring methotrexate in parallel trials (weighted mean difference -4.1 mg per day, 95% confidence interval -6.8 to -1.3) and also in cross-over trials (weighted mean difference -2.9 mg per day, 95% confidence interval -5.9 to -0.2). There was no difference between methotrexate and placebo for forced expiratory volume in one minute (weighted mean difference 0.12 litre, 95% confidence interval -0.21 to 0.45). Hepatotoxicity was a common adverse effect with methotrexate compared to placebo (odds ratio 6.9, 95% confidence interval 3.1 to 15.5). REVIEWER'S CONCLUSIONS: Methotrexate may have a small steroid sparing effect in adults with asthma who are dependent on oral corticosteroids. However, the overall reduction in daily steroid use is probably not large enough to reduce steroid-induced adverse effects. This small potential to reduce the impact of steroid side effects is probably insufficient to offset the adverse effects of methotrexate. PMID- 10796541 TI - Multidisciplinary team interventions for delirium in patients with chronic cognitive impairment. AB - BACKGROUND: Delirium is common in hospitalized elderly people. In the frail elderly, delirium may occur in 60% of those hospitalized. In the cognitively impaired, 45% have been shown to develop delirium and these patients have longer lengths of stay and a higher rate of complications which, amongst other things, together contribute to an increase in cost of care. The combination of being elderly and chronically cognitively impaired leads to a high risk of delirium with the associated increased risk of prolonged hospital stay, complications, and poor outcomes. The management of delirium has commonly been multifaceted - the primary emphasis has always been on the diagnosis and therapy of the precipitating factors, but as this may not be immediately resolved, symptomatic and supportive care may become of major importance. OBJECTIVES: The objective of this review is to assess the available evidence for the effectiveness, if any, of multidisciplinary team interventions in the coordinated care of patients with delirium superimposed on an underlying chronic cognitive impairment compared with the usual care of older cognitively impaired patients. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Cochrane Library, up to and including Issue 1, 1998) was searched using the terms 'delirium, controlled trial, cognitive'. MEDLINE, EMBASE and Psychlit (Ovid via Winspirs up to Feb 1998) were also searched with the same terms. Other sources including personal communications, ongoing trials, conference proceedings, handsearching and reference lists of published papers and books were all searched for relevant randomized controlled trials. The total yield from searching was 157 from which 8 (eight) were retained for consideration in the review. SELECTION CRITERIA: From the initial search yields, all randomised controlled trials involving the management of elderly patients with delirium were identified. A single reviewer (AMB) discarded irrelevant publications based on the title of the publication and its abstract. In the event that the article could possibly be relevant, it was retrieved for further assessment. All references were compiled in a list with a commentary on type of article, eg review, prospective study etc and this was independently considered by the second reviewer (RR) who agreed to review all randomised controlled studies reported on patients with delirium. Selection for possible inclusion in this review was then made on the basis of the participants reported as having chronic cognitive impairment, who then developed incident delirium and were randomly assigned to either coordinated multidisciplinary care or usual care. The outcomes of interest were length of stay in hospital, morbidity (including complications), patient distress & impact on care environment, mortality, discharge arrangements and follow-up including assessment of cognitive function at 6 months. Studies in which patients with chronic cognitive impairment or dementia, managed for incident delirium, according to ICD 9 criteria (see note) were considered eligible for inclusion in the review. Studies of risk factors and non-randomized studies were excluded. Note: this classification has been widely utilised throughout the English speaking medical literature over the past 20 years: ICD 10 is still being incorporated into clinical coding systems and has not been utilised in studies published in 1996. (ABSTRACT TRUNCATED) PMID- 10796542 TI - Educational outreach visits: effects on professional practice and health care outcomes. AB - BACKGROUND: Outreach visits have been identified as an intervention that may improve the practice of health care professionals, in particular prescribing. This type of 'face to face' visit has been referred to as university-based educational detailing, public interest detailing, and academic detailing. OBJECTIVES: To assess the effects of outreach visits on improving health professional practice or patient outcomes. SEARCH STRATEGY: We searched MEDLINE up to March 1997, the Research and Development Resource Base in Continuing Medical Education, and reference lists of related systematic reviews and articles. SELECTION CRITERIA: Randomised trials of outreach visits (defined as a personal visit by a trained person to a health care provider in his or her own setting). The participants were health care professionals. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eighteen studies were included involving more than 1896 physicians. All of the outreach visit interventions consisted of several components, including written materials and conferences. Reminders or audit and feedback complemented some visits. In 13 studies, the targeted behaviours were prescribing practices. In three studies, the behaviours were preventive services, including counselling for smoking cessation. In two studies, the outreach visits were directed toward improving the general management of common problems encountered in general practice, including asthma, diabetes, otitis media, hypertension, anxiety, and acute bronchitis. All studies examined physician behaviour and in three studies other health professionals such as nurses, nursing home attendants or health care workers were targeted. Positive effects on practice were observed in all studies. Only one study measured a patient outcome. Few studies examined the cost effectiveness of outreach. REVIEWER'S CONCLUSIONS: Educational outreach visits, particularly when combined with social marketing, appear to be a promising approach to modifying health professional behaviour, especially prescribing. Further research is needed to assess the effects of outreach visits for other aspects of practice and to identify key characteristics of outreach visits that are important to its success. The cost-effectiveness of outreach visits is not well evaluated. PMID- 10796543 TI - Risperidone versus typical antipsychotic medication for schizophrenia. AB - BACKGROUND: The 'conventional' neuroleptic drugs, such as haloperidol and chlorpromazine, are frequently used as the first line treatment for people with schizophrenia. However, about 5-25% of these people show poor response to these treatments and side effects often makes compliance with the 'older generation' of drug treatment problematic. Although the efficacy of these medications with respect to 'positive' symptoms is well described, little evidence exists that 'conventional' antipsychotic treatment has any effect on the 'negative' symptoms of schizophrenia. Risperidone is one of the 'new generation' neuroleptic compounds. As well as its reputed tendency to cause fewer movement disorders it is claimed that risperidone may improve negative symptoms. OBJECTIVES: To evaluate the effectiveness of risperidone for schizophrenia in comparison to 'conventional' neuroleptic drugs. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1997), Cochrane Schizophrenia Group's Register (1997), The Cochrane Library (1997, Issue1), EMBASE (1980-1997), MEDLINE (1966-1997), PsycLIT (1974-1997), and SCISEARCH (1997) were undertaken. References of all identified studies were searched for further trial citations. Pharmaceutical companies and authors of trials were contacted. SELECTION CRITERIA: All randomised trials comparing risperidone to any 'conventional' neuroleptic treatment for those with schizophrenia or other serious mental illnesses. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were also independently extracted. Sensitivity analyses on dose of risperidone, haloperidol and duration of illness were undertaken for the primary outcomes of clinical improvement, side effects (movement disorders) and acceptability of treatment. For homogeneous dichotomous data the odds ratio (OR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. MAIN RESULTS: Twelve short term studies and two long term studies provided data on 3401 people. This review provides no evidence relating to the effect of risperidone on cognitive or social functioning, quality of life, employment status, discharge from hospital and relapse rates. Risperidone increases the odds of moderate clinical improvement (OR 0.65, CI 0.55-0.77, NNT 10, CI 7-16). It appears to have little or no additional effect on the positive and negative symptoms of schizophrenia but did have less tendency to cause movement disorders, largely in comparison with haloperidol (OR 0.43, CI 0.34-0.55, NNT 7, CI 5-10) for use of antiparkinsonian medication. Risperidone seems to be more acceptable to those with schizophrenia (OR 0.69 CI 0.57-0.83, NNT 15, CI 10-30, 30% baseline risk of dropping out). Those taking risperidone are also marginally less likely to experience somnolence (OR 0.78, CI 0. 61-0.99, NNT 22). Weight gain, however, is more likely with risperidone (OR 1.51 CI 1.14-2.00, NNT 13). Funnel plots show that smaller studies generally show greater benefit for risperidone than larger studies. A publication bias in favour of risperidone amongst the included studies may explain this effect. Sensitivity analyses on dose of risperidone (excluding those receiving 1 or 2 mg) did not materially change the results for the principal outcomes. Excluding data from those on higher doses of haloperidol (>10mg/day) does marginally change the results. Risperidone is less effective in achieving clinical improvement and preventing dropout but outcomes relating to movement disorders change little. REVIEWER'S CONCLUSIONS: Little can be concluded about the long term effects of risperidone and generalising results beyond a comparison with haloperidol would be imprudent. Risperidone may be more acceptable to those with schizophrenia and have marginal benefits in terms of limited clinical improvement and side PMID- 10796544 TI - Selegiline for Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease is the most common cause of dementia in older people accounting for some 60% of cases with late-onset cognitive deterioration. It is now thought that several neurotransmitter dysfunctions are involved from an early stage in the pathogenesis of Alzheimer's disease-associated cognitive decline. The efficacy of selegiline for symptoms of Alzheimer's disease remains controversial and is reflected by its low rate of prescription and the lack of approval by several regulatory authorities in Europe and elsewhere. Reasons for this uncertainty involve the modest overall effects observed in some trials, the lack of benefit observed in several trials, the use of cross-over designs which harbour methodological problems in a disease like dementia and the difficulty in interpreting results from trials when a variety of measurement scales are used to assess outcomes. OBJECTIVES: The objective of this review is to assess whether or not selegiline improves the well-being of patients with Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials, was searched using the terms 'selegiline', 'l-deprenyl', "eldepryl" and "monamine oxidase inhibitor-B". MEDLINE, PsycLIT and EMBASE electronic databases were searched with the above terms in addition to using the group strategy (see group details) to limit the searches to randomised controlled trials. SELECTION CRITERIA: All unconfounded, double-blind, randomised controlled trials in which treatment with selegiline was administered for more than a day and compared to placebo in patients with dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers (JSB & LF), pooled where appropriate and possible, and the weighted or standardised mean differences (95%CI) estimated. Where possible, intention-to-treat data were used but usually the meta analyses were restricted to completers' data (data on people who completed the study). MAIN RESULTS: There are 15 included trials. All trials examined the cognitive effects of selegiline, and in addition, 12 trials examined the behavioural and mood effects. The results of 8 trials suggested some beneficial effect of selegiline in the treatment of cognitive deficits and in 3 trials in the treatment of behaviour and mood. The meta-analysis revealed benefits on memory function as evidenced by improvement in the memory tests from several cognitive tests. Pooling the data for all cognitive tests suggested significant benefits in those subjects treated with selegiline as compared to controls. There were benefits in mood and behaviour as demonstrated by measurements on the Brief Psychiatric Rating Scale and the Dementia Mood Assessment Scale. The global rating scales showed no effect of selegiline. Unfortunately, the evidence using standardised global cognitive scales was extremely limited, for both the MMSE and ADAS-cog. A variety of adverse effects were recorded, but very few patients left a trial as a direct result of the intervention. REVIEWER'S CONCLUSIONS: Although the evidence for a beneficial effect of selegiline on patients with Alzheimer's disease is promising there is not yet enough evidence to recommend its use routinely in practice. The individual patient data review will yield further evidence on the effects of selegiline compared to control as would additional studies evaluating the use of selegiline for the endpoints of standardised cognitive scales, clinician impression of global change, dependency and caregiver quality of life. PMID- 10796545 TI - Support for carers of people with Alzheimer's type dementia. AB - BACKGROUND: Research has highlighted the problems for carers of people with dementia. These include the effective loss of companionship and support of a life partner, social isolation and complex financial, legal and social decision making. The burden of caring is financially, emotionally and physically significant. Studies of the effects of ways of supporting carers and reducing the burden of caring often examine a range of outcomes. Probably because of these characteristics, no formal overview of this area of service provision has been undertaken. Moreover, the term 'intervention' in relation to caregivers of people with Alzheimer's disease is open to wide variations in interpretation at the level of service provision. Interventions range from the application of 'hi-tech' computer technology for socialy isolated carers, to the formation of specialist support groups and respite services. OBJECTIVES: The objective of this review is to provide an assessment of the effectiveness of health and/or social interventions designed to help support the carers of people with Alzheimer's-type dementias. SEARCH STRATEGY: The Cochrane Controlled Trials Register was searched using the terms 'carer*' and 'caregiv*'. See the Cochrane Dementia & Cognitive Impairment group's details for the full search strategy. SELECTION CRITERIA: All randomised trials in which primary carers of people with Alzheimer's disease are allocated to either intervention or non-intervention/control groups and where the intervention was provided by healthcare and/or social services. DATA COLLECTION AND ANALYSIS: Data was extracted by both reviewers and was found to be unsuitable for quantitative synthesis. A qualitative overview is presented and structured according to the comparisons used in the review. Weighted Mean Differences (weighted by sample size) for each outcome and its subcategories are also presented. MAIN RESULTS: The results of the review are inconclusive. No evidence was found for the following comparison interventions: 1) individualised service assessment and planning versus conventional support 2) technology-based carer networking (via computers or telephones) versus conventional support 3) carer education/training versus conventional support 4) multi-faceted/dimensional strategies (such as specialised carer assessment and training) versus conventional support Whilst the overview suggests little or no evidence that interventions to support caregivers of people with Alzheimer's disease are of quantifiable benefit, the poor quality and small sample sizes of the studies, as well as the diversity of interventions examined and outcomes reported, mean that this conclusion cannot be put forward without the need for caution, particularly as some of the studies put forward qualitative evidence which contradicts this conclusion. REVIEWER'S CONCLUSIONS: With the limited nature of the research evidence in mind, it is not possible to recommend either wholesale investment in caregiver support programmes or withdrawal of the same. With the addition of further studies in future updates of this review, expected in early 1999, this presently inconclusive picture may become clearer. A number of conclusions relating to future areas of research can be put forward with more conviction. Specifically, 1) Future trials need to examine interventions included in the existing knowledge base on Alzheimer's carer-supportive interventions. 2) Outcome measures used should mirror those in similar studies. 3) Outcome measures need to be clinically and 'lay' relevant. 4) Trials need to be of longer duration given the 7-10 years median life expectancy of people with Alzheimer's Disease. 5) Sample sizes need to be increased and calculated properly given the likelihood of a moderate intervention effect (if any). 7) Blinding at the outcome assessment stage needs to be a part of future basic trial designs if bias is to be avoided. 8) If reviews are to have their power increased then quality o PMID- 10796546 TI - Neuroleptic reduction and/or cessation and neuroleptics as specific treatments for tardive dyskinesia. AB - BACKGROUND: Since the 1950s neuroleptic medication has been extensively used to treat people with chronic mental illnesses, such as schizophrenia. These drugs, however, have been also associated with a wide range of adverse effects, including movement disorders such as tardive dyskinesia (TD). Various strategies have been examined to reduce a person's cumulative exposure to neuroleptics. These studies include dose reduction, intermittent dosing strategies, such as drug holidays, and neuroleptic cessation. OBJECTIVES: To determine whether, for those people with both schizophrenia (or other chronic mental illnesses) and tardive dyskinesia (TD), a reduction or cessation of neuroleptic drugs was associated with reduction in TD symptoms. A secondary objective was to determine whether the use of specific neuroleptics for similar groups of people could be a treatment for already established TD. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1997), Cochrane Schizophrenia Group's Register of trials (1997), EMBASE (1980-1997), LILACS (1982-1996), MEDLINE (1966-1997), PsycLIT (1974-1997), and SCISEARCH (1997) were undertaken. References of all identified studies were searched for further trial citations. Principal authors of trials were contacted. SELECTION CRITERIA: Reports were included if they assessed the treatment of neuroleptic-induced tardive dyskinesia in people with schizophrenia or other chronic mental illnesses and already established TD, who had been randomly allocated to (a) neuroleptic cessation (placebo or no intervention) versus neuroleptic maintenance; b. neuroleptic reduction (including intermittent strategies) versus neuroleptic maintenance; or c. specific neuroleptics for the treatment of TD versus placebo or no intervention. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently and the Odds Ratio (95% CI) or the average difference (95% CI) were estimated. The reviewers assumed that people who dropped out had no improvement. MAIN RESULTS: Two trials were able to be included in this review. Sixty two were excluded and 16 are awaiting assessment. Seven trials are still pending classification. No randomised controlled trial-derived data were available to clarify the role of neuroleptics as treatments for TD. This includes the atypical antipsychotics including clozapine. Despite neuroleptic cessation being a frequently first-line recommendation, there were no RCT-derived data to support this. Two studies ( approximately approximately Cookson 1987 approximately approximately , approximately approximately Kane 1983 approximately approximately ) found a reduction in TD associated with neuroleptic reduction. REVIEWER'S CONCLUSIONS: The lack of evidence to support the efficacy of neuroleptic cessation as a treatment for TD, combined with the accumulating evidence of an increased risk of relapse should antipsychotic drugs be reduced, makes this intervention a hazardous treatment for TD. Dose reduction may offer some benefit as a treatment for TD compared to standard levels of neuroleptic use. There is a need to evaluate the utility of clozapine and the 'atypical' antipsychotics as treatments for established TD. PMID- 10796547 TI - Thioridazine for dementia. AB - BACKGROUND: Neuroleptic drugs are controversial treatments in dementia, with evidence accumulating that they may hasten clinical decline. Despite these concerns, they are commonly prescribed for elderly and demented patients. Thioridazine, a phenothiazine neuroleptic, is one of the most commonly prescribed. It has often been a preferred agent because it is thought to produce relatively less frequent motor side effects. The drug has significant sedative effects, and it is thought that these are the main mechanism of action in calming and controlling the patient. However, pharmacologically, it also has marked anticholinergic properties that could potentially have a detrimental effect on cognitive function. OBJECTIVES: To determine the evidence on which the use of thioridazine in dementia is based in terms of: 1) efficacy in controlling symptoms 2) cognitive outcome for the patient 3) safety SEARCH STRATEGY: The Cochrane Controlled Trials Register and other electronic databases were searched using the terms 'thioridazine', 'melleril', 'dementia' and 'old age'. In addition, Novartis, the pharmaceutical company that developed and markets thioridazine, was approached and asked to release any published or unpublished data they had on file. SELECTION CRITERIA: Unconfounded, single-blind or double blind, randomised trials were identified in which treatment with thioridazine was administered for more than one dose and compared to an alternative intervention in patients with dementia of any aetiology. Trials in which allocation to treatment or comparator were not truly random, or in which treatment allocation was not concealed were reviewed but are not included in the data analysis. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers (VC, CAK and RJH). For continuous and ordinal variables, the main outcome measures of interest were the final assessment score and the change in score from baseline to the final assessment. The assessment scores were provided by behavioural rating scales, clinical global impression scales, functional assessment scales, psychometric test scores, and frequency and severity of adverse events. Data were pooled where appropriate or possible, and the Peto odds ratio (95%CI) or the weighted mean difference (95%CI) estimated. Where possible, intention to treat data were used. MAIN RESULTS: The meta-analysis showed that, compared with placebo, thioridazine reduced anxiety symptoms as evidenced by changes on the Hamilton Anxiety Scale. However, there was no significant effect on clinical global change, and a non-significant trend for higher adverse effects with thioridazine. Compared to diazepam, thioridazine was superior in terms of some anxiety symptoms, with similar adverse effects. Global clinical evaluation scales mostly did not favour either treatment. Compared to chlormethiazole, thioridazine was significantly inferior when assessed on some items of the CAPE and the Crichton Geriatric Behavioural Rating Scales. Thioridazine was also associated with significantly more dizziness. No superiority for thioridazine was shown in comparisons with etoperidone, loxapine or zuclopenthixol. REVIEWER'S CONCLUSIONS: Very limited data are available to support the use of thioridazine in the treatment of dementia. If thioridazine were not currently in widespread clinical use, there would be inadequate evidence to support its introduction. The only positive effect of thioridazine when compared to placebo is the reduction of anxiety. When compared to placebo, other neuroleptics, and other sedatives it has equal or higher rates of adverse effects. Clinicians should be aware that there is no evidence to support the use of thioridazine in dementia, and its use may expose patients to excess side effects. PMID- 10796548 TI - Barrier agents for preventing adhesions after surgery for subfertility. AB - BACKGROUND: Pelvic adhesions can be the result of inflamation, endometriosis or surgical trauma. Prevention of postoperative adhesions (either new or reoccurance) has been postulated by using barriers to prevent two surfaces being in contact. When pelvic surgery is being undertaken strategies to reduce pelvic adhesions occurring may be undertaken and these include barrier agents which are placed between the pelvic structures. Two synthetic barriers with differential characteristics are commercially available: oxidised regenerated cellulose (Interceed) and polytetrafluoroethylene (PTFC) (GoreTex). OBJECTIVES: The objective of this review was to assess the effect of mechanical barriers (materials interposed between pelvic structures to prevent adherence of serosal surfaces) used during pelvic surgery in women of reproductive age on pregnancy rates, pelvic pain, or postoperative adhesion reformation. SEARCH STRATEGY: The Cochrane Menstrual Disorders and Subfertility Group specialised register of controlled clinical trials was undertaken. In addition, companies were contacted for unpublished trials. SELECTION CRITERIA: Randomised controlled trials or controlled clinical trials of barriers versus no treatment or other barriers in women undergoing fertility preserving pelvic surgery. DATA COLLECTION AND ANALYSIS: Reviewers assessed eligibility and trial quality. MAIN RESULTS: 15 randomised controlled trials were included. Five trials randomised patients while the remainder randomised pelvic organs. Laparoscopy was the primary surgical technique in six trials while the remaining trials were laparotomy. Indications for surgery included myomectomy (five trials), ovarian surgery (four trials), pelvic adhesions (six trials), endometriosis (two trials) and mixed (one trial). Thirteen trials assessed Interceed versus no treatment, two assessed Interceed versus Gore-Tex, one trial assessed Gore-Tex versus no treatment, and one trial assessed Seprafilm versus no treatment. No study reported pregnancy or reduction in pain as an outcome. The use of Interceed in women was associated with reduced incidence of pelvic adhesion formation, both new formation and re-formation following laparoscopic surgery and after laparotomy. Gore-Tex was more effective than no barrier or Interceed in preventing adhesion formation. There was limited evidence that Seprafilm was effective in preventing adhesion formation in women following myomectomy. REVIEWER'S CONCLUSIONS: The absorbable adhesion barrier Interceed reduces the incidence of adhesion formation, both new formation and re formation, at laparoscopy and laparotomy, but there are insufficient data to support its use to improve pregnancy rates. Gore-Tex may be superior to Interceed in preventing adhesion formation but its usefulness is limited by the need for suturing and later removal. There was no evidence of effectiveness of Seprafilm in preventing adhesion formation. PMID- 10796549 TI - Zuclopenthixol acetate in the treatment of acute schizophrenia and similar serious mental illnesses. AB - BACKGROUND: People with schizophrenia or other psychotic illnesses may have delusions or hallucinations that may lead them to be aggressive or violent to themselves or others. Medication that is used in this context require the properties of rapid onset of effect (tranquillisation or at least initial sedation in order to quell aggressive or disorganised behaviour, but also antipsychotic effect), low frequency of administration and low levels of side effects, such as cardiological or neurological side effects, or pain at the injection site. Zuclopenthixol is the cis(Z)-isomer of clopenthixol, a neuroleptic of the thioxanthene group, used for treating people with psychotic symptoms. There is one oral preparation and two depot forms: zuclopenthixol acetate and zuclopenthixol decanoate. The acetate version does not stay in the body for very long (a single dose persists for only 72 hours) and is said to have these properties. OBJECTIVES: To estimate the effectiveness of zuclopenthixol acetate for the acute treatment of serious mental illnesses in comparison to other neuroleptic drugs. SEARCH STRATEGY: Searches of the Cochrane Schizophrenia Group's Register of Trials, The Cochrane Library, MEDLINE, abstracts of congresses and trial reference lists were performed. Appeals for unpublished data to the research community and to the Medical Information Department of Lundbeck Limited were also made. Attempts were made to contact relevant authors. SELECTION CRITERIA: Two reviewers independently assessed citations or papers and selected all randomised trials that included people with serious mental illnesses and compared zuclopenthixol acetate with other drug regimes. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Attempts were made to contact authors for additional or missing information. Odds-ratios (OR) and 95% confidence intervals (CI) were estimated for binary data. Where possible, OR were pooled using Peto method and intention-to-treat analysis undertaken. Mean differences were used for continuous variables. MAIN RESULTS: Pooled data shows no difference for the outcome 'no important improvement' in psychotic symptoms at the end of the follow-up period (OR 0.84 CI 0. 34-2.05). Sedation was evaluated using different instruments. Only one study presented data which suggested an earlier and more intense sedation in zuclopenthixol acetate users at 4 hours (OR 0.18 CI 0. 04-0.93). Use of additional antipsychotic medication was not avoided in the zuclopentixol acetate group (OR 2.18, CI 0.64-7.42) and data on total number of administrations was not obtainable. Side effect data were poorly reported but there is no evidence of a consistent difference between zuclopenthixol acetate and other 'standard drugs' for either the pattern of side effects or the wish to leave the study early. Hospital and service outcomes, number of aggressive incidents, satisfaction with care and economic outcomes were not addressed by any study. REVIEWER'S CONCLUSIONS: Recommendations on the use of zuclopenthixol acetate for the management of psychiatric emergencies in preference to 'standard' treatment have to be viewed with caution. Most trials present important methodological flaws and findings are poorly reported. This review did not find any suggestion that zuclopenthixol acetate is more effective in controlling aggressive/disorganised behaviour, acute psychotic symptoms, or preventing side effects. There were no data directly related to tranquillisation, but it may produce more earlier and intense sedation than oral haloperidol. Well conducted randomized controlled trials are needed to confirm claims related to the use of zuclopenthixol acetate in emergency psychiatry. PMID- 10796550 TI - Interventions for treating tuberculous pericarditis. AB - BACKGROUND: Tuberculous (TB) pericarditis is becoming more common. The infection can result in fluid around the heart, which can be fatal. OBJECTIVES: To evaluate evidence from trials about the effects of medical and surgical treatments for TB pericarditis on death and life-threatening conditions. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, the Cochrane controlled trials register, Medline, Embase and reference lists of articles; contact with experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of treatments for TB pericarditis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Meta-analysis using fixed effects models calculated summary statistics, provided there was no significant heterogeneity, and expressed results as relative risk. MAIN RESULTS: Three trials met the inclusion criteria, with a total of 411 participants. Treatments were adjuvant steroids and surgical drainage. Two small trials tested steroids. There were fewer deaths (all causes) in the intervention group, but the numbers were small and the result could have occurred by chance (relative risk [RR] 0.65, 95% confidence interval [CI] 0.36 to 1.16, n = 350). In one trial studying patients with effusion, "cure" was higher in the steroid group (alive and free of disability at 2 years (RR 0.69, 95% CI 0.29 to 0.80, n = 221). One trial examined open surgical drainage compared with conservative management, and showed no impact of surgery on death, but a protective effective against cadiac tamponade (RR 0.04, 95% CI 0.00 to 0.64). REVIEWER'S CONCLUSIONS: Steroids have potentially large impacts on survival, but trials are too small to test this. We believe further placebo controlled trials of steroids are warranted, exploring whether the presence of effusion or fibrosis modifies effects. Surgical options also require further evaluation. PMID- 10796551 TI - Artemisinin derivatives for treating severe malaria. AB - BACKGROUND: Artemisinin derivatives may have advantages over quinoline drugs for treating severe malaria since they are fast acting and effective against quinine resistant malaria parasites. OBJECTIVES: The objective of this review was to assess the effects of artemisinin drugs for severe and complicated falciparum malaria in adults and children. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, Cochrane Controlled Trials Register, Medline, Embase, Science Citation Index, Lilacs, African Index Medicus, conference abstracts and reference lists of articles. We contacted organisations, researchers in the field and drug companies. SELECTION CRITERIA: Randomised and pseudo-randomised trials comparing artemisinin drugs (rectal, intramuscular or intravenous) with standard treatment, or comparisons between artemisinin derivatives in adults or children with severe or complicated falciparum malaria. DATA COLLECTION AND ANALYSIS: Eligibility, trial quality assessment and data extraction were done independently by two reviewers. Study authors were contacted for additional information. MAIN RESULTS: Twenty three trials are included, allocation concealment was adequate in nine. Sixteen trials compared artemisinin drugs with quinine in 2653 patients. Artemisinin drugs were associated with better survival (mortality odds ratio 0.61, 95% confidence interval 0.46 to 0.82, random effects model). In trials where concealment of allocation was adequate (2261 patients), this was barely statistically significant (odds ratio 0.72, 95% CI 0.54 to 0.96, random effects model). In 1939 patients with cerebral malaria, mortality was also lower with artemisinin drugs overall (odds ratio 0.63, 95% CI 0.44 to 0.88, random effects model). The difference was not significant however when only trials reporting adequate concealment of allocation were analysed (odds ratio 0.78, 95% CI 0.55 to 1.10, random effects model) based on 1607 patients. No difference in neurological sequelae was shown. Compared with quinine, artemisinin drugs showed faster parasite clearance from the blood and similar adverse effects. REVIEWER'S CONCLUSIONS: The evidence suggests that artemisinin drugs are no worse than quinine in preventing death in severe or complicated malaria. No artemisinin derivative appears to be better than the others. PMID- 10796552 TI - Interventions for treating schistosomiasis mansoni. AB - BACKGROUND: Schistosomiasis is a parasite that is carried by freshwater snails. The intestinal form infects the intestine, liver and spleen and can be fatal. OBJECTIVES: The objective of this review was to assess the effects of oxamniquine or praziquantel for treating Schistosomiasis mansoni SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Lilacs and reference lists of articles. The Revista da Sociedade Brasileira de Medicina Tropical and Brazilian Tropical Medicine Congress abstracts were handsearched SELECTION CRITERIA: Randomised and quasi randomised trials comparing oxamniquine and/or praziquantel to placebo for the treatment of Schistosomiasis mansoni. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Thirteen trials met the inclusion criteria. Praziquantel and oxamniquine were effective in curing Schistosoma mansoni infection when compared to placebo. In Africa, praziquantel 40 mg/Kg is more effective than oxamniquine 15 mg/Kg in individuals older than 14 years (OR 3.54, 95%CI 1.70, 7.38), but no difference was found when compared with oxamniquine 30 mg/Kg (OR 0.29, 95%CI 0.08, 1.01). In Brazil, praziquantel was equally effective when compared with oxamniquine in individuals older than 14 years (OR 1.70, 95%CI 0.83, 3.49). Both drugs appear safe. There was no difference in reinfection rate between zinc supplementation and placebo (OR 0.82, 95%CI 0.47, 1.41). REVIEWER'S CONCLUSIONS: IPraziquantel and oxamniquine both appear to be effective for the treatment of Schistosomiasis mansoni, although lower doses of oxamniquine (less than 30 milligrams per kilogram) may not be as effective. PMID- 10796553 TI - Echinacea for preventing and treating the common cold. AB - BACKGROUND: Extracts of the plant Echinacea (family Compositae) are widely used in some European countries and the USA for upper respiratory tract infections. OBJECTIVES: The objective of this review was to assess the effects of preparations containing extracts of Echinacea in the prevention and treatment of the common cold. SEARCH STRATEGY: We searched the Cochrane Acute Respiratory Infections Group and Complementary Medicine Field's trials registers, MEDLINE, EMBASE, Phytodok and reference lists of articles. We also contacted researchers and manufacturers. Date of last search: Spring 1998. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing preparations containing an extract of Echinacea compared with a placebo, no treatment, or another treatment for common colds. DATA COLLECTION AND ANALYSIS: At least two independent reviewers assessed trial quality and extracted data. MAIN RESULTS: Sixteen trials (eight prevention trials, and eight trials on treatment of upper respiratory tract infections) with a total of 3396 participants were included. Variation in preparations investigated and methodological quality of trials precluded quantitative meta-analysis. Overall, the results suggested that some Echinacea preparations may be better than placebo. REVIEWER'S CONCLUSIONS: The majority of the available studies report positive results. However there is not enough evidence to recommend a specific Echinacea product, or Echinacea preparations for the treatment or prevention of common colds. PMID- 10796554 TI - Diabetes care: the effectiveness of systems for routine surveillance for people with diabetes. AB - BACKGROUND: There is wide variation in the extent of general practice involvement in diabetes care. OBJECTIVES: To assess the effects of involving primary care professionals in the routine review and surveillance for complications of people with established diabetes mellitus compared with secondary care specialist follow up. SEARCH STRATEGY: We searched the Cochrane Diabetes Group specialised register, The Cochrane Library, MEDLINE (January 1966 to December 1996), EMBASE (to December 1996), Cinahl (to December 1996), National Research Register (to December 1996), PsycLIT (to December 1996), HealthSTAR (to December 1996), CRIB (to December 1996), Dissertation Abstracts (to December 1996), and reference lists of articles. SELECTION CRITERIA: Randomised trials in which people with diabetes were allocated to a system of review and surveillance for complications by primary care professionals. Outcomes included mortality, metabolic control, cardiovascular risk factors, quality of life, functional status, satisfaction, hospital admissions, costs, completeness of screening, and development of complications. DATA COLLECTION AND ANALYSIS: The reviewer assessed trial quality and extracted data. Analysis was on an intention to treat basis. General practice care was categorised into routine or prompted care and a stratified analysis undertaken. MAIN RESULTS: Five trials involving 1058 people were included. Results were heterogeneous between trials. In those schemes featuring more intensive support through a prompting system for general practitioners and patients, there was no difference in mortality between hospital and general practice care (odds ratio 1.06, 95% confidence interval 0.53 to 2.11), HbA1 tended to be lower (a weighted difference in means of -0.27%, 95% confidence interval -0.59 to 0.03) and losses to follow up were significantly lower (odds ratio 0.37, 95% confidence interval 0.22 to 0.61) in primary care. However, schemes with less well-developed support for family doctors were associated with adverse outcomes for patients. Quality of life, cardiovascular risk factors, functional status and the development of complications were infrequently assessed. REVIEWER'S CONCLUSIONS: Unstructured care in the community is associated with poorer follow up, greater mortality and worse glycaemic control than hospital care. Computerised central recall, with prompting for patients and their family doctors, can achieve standards of care as good or better than hospital outpatient care, at least in the short term. The evidence supports provision of regular prompted recall and review of people with diabetes by willing general practitioners and demonstrates that this can be achieved, if suitable organisation is in place. PMID- 10796555 TI - Oral 5-aminosalicylic acid for inducing remission in ulcerative colitis. AB - OBJECTIVES: To assess the efficacy, dose-responsiveness and safety of the newer release formulations of 5-aminosalicylic acid (5-ASA) compared to placebo or sulfasalazine (SASP) for the induction of remission in active ulcerative colitis. SEARCH STRATEGY: A computer-assisted literature search for relevant studies (1981 1998) was performed using MEDLINE, BIOS, the Cochrane Controlled Trials Register and the Science Citation Index, followed by a manual search of reference lists from previously retrieved articles, review articles, symposia proceedings, and abstracts from major gastrointestinal conferences. SELECTION CRITERIA: Studies were accepted for analysis if they were randomized, double-blinded, and controlled clinical trials of parallel design, with treatment durations of a minimum of four weeks. DATA COLLECTION AND ANALYSIS: Based on an intention to treat principle, the outcomes of interest in the treatment of active disease were the failure to induce global/clinical remission, global/clinical improvement, endoscopic remission, or endoscopic improvement. MAIN RESULTS: 5-ASA was superior to placebo with regard to all measured outcome variables. For the failure to induce global/clinical improvement or remission, the pooled Peto odds ratio was 0.51 (95% CI, 0.35 to 0.76). A dose-response trend for 5-ASA was also observed. When 5-ASA was compared to SASP, the pooled Peto odds ratio was 0.87 (CI, 0.63 to 1.21) for the failure to induce global/clinical improvement or remission, and 0.66 (CI, 0.42 to 1.04) for the failure to induce endoscopic improvement. SASP was not as well tolerated as 5-ASA. REVIEWER'S CONCLUSIONS: The newer 5-ASA preparations were superior to placebo and tended towards therapeutic benefit over SASP. However, considering their relative costs, a clinical advantage to using the newer 5-ASA preparations in place of SASP appears unlikely. PMID- 10796556 TI - Oral 5-aminosalicylic acid for maintaining remission in ulcerative colitis. AB - OBJECTIVES: To assess the efficacy, dose-responsiveness and safety of the newer release formulations of 5-aminosalicylic acid (5-ASA) compared to placebo or sulfasalazine (SASP) in the maintenance of remission in ulcerative colitis. SEARCH STRATEGY: A computer-assisted literature search for relevant studies (1981 1998) was performed using MEDLINE, BIOS, the Cochrane Controlled Trials Register, the Inflammatory Bowel Disease Trials Register, and Science Citation Index, followed by a manual search of reference lists from previously retrieved articles, review articles, symposia proceedings, and abstracts from major gastrointestinal conferences. SELECTION CRITERIA: Studies were accepted for analysis if they were prospective, randomized, double-blinded, and placebo- or SASP-controlled clinical trials of parallel design with treatment duration of at least six months. DATA COLLECTION AND ANALYSIS: Based on an intention to treat principle, the primary outcome was the failure to maintain clinical or endoscopic remission. Secondary outcomes were the number of patients experiencing adverse events, the number of patients withdrawn due to adverse events, and exclusions or withdrawals after entry into the study (not due to relapse). All data were analyzed using the Peto odds ratio and corresponding 95% confidence intervals (CI). MAIN RESULTS: The Peto odds ratio for the failure to maintain clinical or endoscopic remission (withdrawals and relapses) for 5-ASA versus placebo was 0. 47 (95% CI, 0.36 to 0.62) with an NNT of 6. These values were also calculated for the trials in which SASP and 5-ASA were compared, revealing an odds ratio of 1.29 (95% CI, 1.05 to 1.57), with a negative NNT value (-19), suggesting a higher degree of therapeutic effectiveness for SASP. SASP and 5-ASA had similar adverse event profiles, with odds ratios of 1.16(0.62 to 2.16), and 1.31(0.86 to 1.99), respectively. The NNH values were determined to be 171 and 78 respectively. REVIEWER'S CONCLUSIONS: The newer 5-ASA preparations were superior to placebo in maintenance therapy. However, the newer preparations had a statistically significant therapeutic inferiority relative to SASP. PMID- 10796557 TI - Azathioprine or 6-mercaptopurine for inducing remission of Crohn's disease. AB - OBJECTIVES: To determine the effectiveness of azathioprine and 6-mercaptopurine in inducing remission of active Crohn's disease. SEARCH STRATEGY: Studies were selected using the MEDLINE data base (1966 - December 1997), abstracts from major gastrointestinal meetings and references from published articles and reviews. The Cochrane Controlled Trials Register and the Inflammatory Bowel Disease Review Group Trials Register was also searched. SELECTION CRITERIA: Eight randomized placebo controlled trials of azathioprine and 6-mercaptopurine therapy in adult patients were identified: five dealt with active disease and three had multiple therapeutic arms. DATA COLLECTION AND ANALYSIS: Data were extracted by three independent observers based on the intention to treat principle. Each study was given a quality score based on predetermined criteria. Extracted data were converted to 2X2 tables (response versus no response and antimetabolite versus placebo) and then synthesized into a summary test statistic using the pooled odds ratio and 95% confidence intervals as described by Cochran and Mantel and Haenszel ('Odds Ratio' in MetaView). MAIN RESULTS: The odds ratio of a response to azathioprine or 6-mercaptopurine therapy compared with placebo in active Crohn's disease was 2.36 (95% CI 1.57-3.53). This corresponded to a number needed to treat of about 5 to observe an effect of therapy in one patient. When the two trials using 6-mercaptopurine in active disease were excluded from the analysis, the odds ratio of response was 2.04 (CI 1.24 - 3.35). Treatment >/= 17 weeks increased the odds ratio of a response to 2.51 (CI 1.63-3. 88). A steroid sparing effect was seen with an odds ratio of 3.86 (CI 2.14 - 6.96), corresponding to a number needed to treat of about 3 to observe steroid sparing in one patient. Adverse events requiring withdrawal from a trial, principally allergy, leukopenia, pancreatitis, and nausea were increased on therapy with an odds ratio of 3.01 (CI 1.30 - 6.96). The number needed to treat to observe one adverse event in one patient treated with azathioprine or 6-mercaptopurine was 14. REVIEWER'S CONCLUSIONS: Azathioprine and 6-mercaptopurine are effective therapy for inducing remission in active Crohn's disease. The odds ratio of response increases after >/= 17 weeks of therapy, suggesting that there is a minimum length of time for a trial of azathioprine or 6-mercaptopurine therapy. Adverse events were more common among patients on therapy. PMID- 10796558 TI - Aversive smoking for smoking cessation. AB - BACKGROUND: Aversion therapy pairs the pleasurable stimulus of smoking a cigarette with some unpleasant stimulus. The objective is to extinguish the urge to smoke. OBJECTIVES: This review has two aims. First, to determine the efficacy of rapid smoking and other aversive methods in helping smokers stop smoking. Second, to determine whether there is a dose-response effect on smoking cessation at different levels of aversive stimulation. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register for studies which evaluated any technique of aversive smoking. SELECTION CRITERIA: Randomised trials which compared aversion treatments with 'inactive' procedures or which compared aversion treatments of different intensity for smoking cessation. Trials must have reported follow-up of least 6 months from beginning of treatment. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the study population, the type of aversion treatment, the outcome measure, method of randomisation and completeness of follow-up. The outcome measure was abstinence from smoking at maximum follow-up, using the strictest measure reported by the authors. Subjects lost to follow-up were regarded as smokers. Where appropriate, we performed meta analysis using a fixed effects model. MAIN RESULTS: Twenty four trials met the inclusion criteria. Ten included rapid smoking and ten used other aversion methods. Ten trials included two or more conditions allowing assessment of a dose response to aversive stimulation. The odds ratio for abstinence following rapid smoking compared to control was 2.08 (95% confidence interval 1.39 to 3.12). Several factors suggest that this finding should be interpreted cautiously. A funnel plot of included studies was asymmetric, due to the relative absence of small studies with negative results. Most trials had a number of serious methodological problems likely to lead to spurious positive results. The only trial using biochemical validation of all self reported cessation gave a non significant result. Other aversion methods were not shown to be effective (odds ratio 1.19, 95% confidence interval 0.77 to 1.83). There was a borderline dose response to the level of aversive stimulation (odds ratio 1.66, 95% confidence interval 1.00 to 2.78). REVIEWER'S CONCLUSIONS: The existing studies provide insufficient evidence to determine the efficacy of rapid smoking, or whether there is a dose-response to aversive stimulation. Milder versions of aversive smoking seem to lack specific efficacy. Rapid smoking is an unproven method with sufficient indications of promise to warrant evaluation using modern rigorous methodology. PMID- 10796559 TI - Newer atypical antipsychotic medication versus clozapine for schizophrenia. AB - BACKGROUND: Clozapine is an atypical antipsychotic drug, which is claimed to have superior efficacy and to cause fewer movement disorders. However, clozapine carries a significant risk of serious blood disorders. Newer atypical antipsychotics are safer alternatives that might share the benefits of clozapine. It is thus of interest to compare the effectiveness of newer atypical antipsychotics with the effectiveness of clozapine. OBJECTIVES: To evaluate the clinical effects of newer atypical antipsychotic drugs in comparison to clozapine for schizophrenia. SEARCH STRATEGY: Publications in all languages were searched from the following databases: Biological Abstracts/BIOSIS (1980-1999), The Cochrane Schizophrenia Group's Register of Trials (1998), The Cochrane Library CENTRAL Register (Issue 4, 1999), EMBASE (1980-1998), MEDLINE (1966-1999), LILACS/CD-ROM (1998), and PsycLIT/PsycINFO (1974-1999). Trials were also sought from recent conference proceedings and reference lists of included papers. Authors of recent trials and the manufacturers of clozapine, iloperidone, olanzapine, quetiapine, remoxipride, risperidone, sertindole, ziprasidone and zotepine were contacted. SELECTION CRITERIA: All randomised controlled trials comparing clozapine with newer atypical antipsychotic drugs were included by independent assessment by two reviewers. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Relative risks (RR) and 95% confidence intervals (CI) of homogenous dichotomous data were calculated. A random effects model was used for heterogeneous dichotomous data. Where possible the number needed to treat (NNT) statistic with 95%CI were also calculated. Weighted or standardised means were calculated for continuous data. Due to the small number of included studies, sensitivity analyses or funnel plot statistics were not undertaken for this version of the review. MAIN RESULTS: The current review includes eight studies (22 papers), of which three studies are 4-6 weeks in duration and only one study is of more than 12 weeks' duration. Newer atypical drugs seemed to be broadly similar to clozapine using a clinical global index or trialists' definitions of improvement, but this result was obtained from a relatively small number of studies. Due to the small number of studies and patients, wide confidence intervals were seen when their effectiveness as measured by symptom rating scales was compared. Social functioning was better in patients on newer atypical medication (risperidone) than in those on clozapine, but this finding is based on a single underpowered trial and has to be interpreted with caution. Clozapine and newer atypical drugs showed their adverse effect profile to be dissimilar: while clozapine produced more fatigue, hypersalivation, nausea, and orthostatic dizziness, new atypical drugs, with the exception of olanzapine, produced more extrapyramidal symptoms. The impact of these drugs and their effects on patients' day-to-day quality of life, service use, hospital admission, and pharmacoeconomics was not measured. REVIEWER'S CONCLUSIONS: The equal effectiveness and tolerability of new atypical drugs in comparison with clozapine is not yet demonstrated. Lack of statistical power to determine the comparative efficacy and effectiveness of newer atypical drugs makes it difficult to judge whether newer drugs are more effective, less effective or equivalent. Trials of sufficient power, with longer duration, measuring clinically important outcomes, are needed to assess the true comparative clinical effectiveness, tolerability and cost effectiveness of newer drugs in relation to clozapine. PMID- 10796560 TI - Quetiapine for schizophrenia. AB - BACKGROUND: Quetiapine is a novel atypical antipsychotic with low propensity for movement disorder adverse effects. It is used for treatment of schizophrenia and other psychoses. OBJECTIVES: To determine the effects of quetiapine for schizophrenia in comparison to placebo, classical and other atypical antipsychotics. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1997), CINAHL (1982-1997), the Cochrane Library (1998, Issue 1), the Cochrane Schizophrenia Group's Register of trials (1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1974-1997), SocioFile (1974-1997) and many conference proceedings and hand searches of specific journals were undertaken. Zeneca Pharmaceuticals was contacted for information regarding unpublished trials. SELECTION CRITERIA: All controlled trials where adults with schizophrenia or similar illnesses were randomised to quetiapine, placebo or other neuroleptic drugs and where clinically relevant outcomes were reported. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were also independently extracted. For homogeneous dichotomous data the Peto odds ratio (OR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) was calculated on an intention-to-treat basis. MAIN RESULTS: Seven trials of short duration are included (31 reports) and seven are excluded (15 reports). Apart from that of 'leaving the study early', all other results may be prone to bias and should be viewed with caution since dropout rates are high (48 61%) in each arm of all studies. There are data suggesting less people allocated quetiapine leave the study early (53%) than those in the placebo group (61%) (OR 0.67 CI 0.48-0. 95). Data incorporating considerable assumptions about the many people who left early suggest that global state and psychotic symptoms - both positive and negative - may be more helped by quetiapine than placebo. Although some of these data reach statistical significance their clinical importance is difficult to interpret. While the incidences of extrapyramidal side effects are not different between quetiapine and placebo, side effects such as dizziness and dry mouth are more prevalent in the quetiapine treated group. High proportions of trial participants also leave when quetiapine is compared to chlorpromazine or haloperidol (57% by six weeks). Quetiapine is as potent as chlorpromazine and haloperidol as regards global and mental state but it may cause higher incidences of dry mouth and sleepiness. Extrapyramidal side effects are the same as those of chlorpromazine but may be less than haloperidol. High dose quetiapine is better than low dose quetiapine with regard to leaving the study early, and limited data suggest that the higher dose is also better at marginally improving global state (n = 1, OR 0.70, CI 0.50-0.99, NNT 11). There are no clear differences between high and low dose groups in respect of extrapyramidal side effects. REVIEWER'S CONCLUSIONS: The high dropout rates are a large problem in interpreting any results other than 'leaving the study early' since about half the data were not available at the end of studies. Before quetiapine's use can be recommended, we need more large, well conducted trials that provide short, medium and long term outcomes relevant to carers and clinicians. PMID- 10796561 TI - Fully intermittent dosing with drugs for tuberculosis. AB - BACKGROUND: The number of people infected with tuberculosis continues to rise world-wide. Rifampicin-containing treatment regimens can achieve high cure rates. Intermittent drug treatment delivered in the community has the potential to improve adherence to treatment. OBJECTIVES: The objective of this review was to compare the effectiveness of rifampicin-containing short-course chemotherapy regimens, given two or three times a week, with similar regimens given daily in patients with pulmonary tuberculosis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of any multi drug regimen containing rifampicin in patients with confirmed pulmonary tuberculosis. Treatment had to be given up to three times a week for up to nine months, with any initial daily dosing period not more than one month, and was compared to daily dosing throughout for the same period. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality. MAIN RESULTS: One trial involving 399 patients was included. The trial compared treatment three times per week with daily treatment for six months. There was no difference in cure rate (198 out of 199 people in the intermittent group compared to all 200 in the daily group), but 5 patients relapsed in the group receiving intermittent therapy compared to one in the group receiving the daily regimen. REVIEWER'S CONCLUSIONS: There is not enough evidence to assess the equivalence of effect between fully intermittent, rifampicin-containing short-course chemotherapy and similar daily therapy in patients with pulmonary tuberculosis. Larger randomised studies are required to establish the effectiveness of fully intermittent, short-course chemotherapy. PMID- 10796562 TI - Steroids for treating cerebral malaria. AB - BACKGROUND: Cerebral malaria is associated with swelling of the brain. Corticosteroid drugs could reduce the harmful effects of this swelling, but could also suppress host immunity to infection. OBJECTIVES: This review aimed to assess the effects of steroids in patients with cerebral malaria in relation to survival and long-term disability. SEARCH STRATEGY: The Cochrane Controlled Trials Register, the Cochrane Infectious Diseases Group Trials Register, existing reviews, and contact with experts in the field. SELECTION CRITERIA: Randomised controlled trials comparing steroids and no steroids in otherwise identical treatments for patients with cerebral malaria. DATA COLLECTION AND ANALYSIS: Assessments of inclusion criteria, trial quality and data extraction were done by each author independently. The outcomes sought were death, death with life threatening complications, other complications, and disability. MAIN RESULTS: Two trials met the inclusion criteria. Out of a total of 143 patients, there were 30 deaths, distributed evenly between the steroid and the comparator group. The researchers reported clinical complications as the number of events in each trial arm, and did not exclude complications occurring in fatalities. This makes it difficult to interpret the reports of significantly more episodes of gastro intestinal bleeding and seizures in the steroid group. No studies examined disability. REVIEWER'S CONCLUSIONS: There is currently no evidence of benefit from steroids in this condition, but the small number of participants means it is difficult to exclude an effect on mortality in either direction. Data on clinical complications are difficult to assess. PMID- 10796563 TI - Vaccines for preventing cholera. AB - BACKGROUND: Oral cholera vaccines (either killed whole cell or live recombinant vaccines) are newer alternatives to the parenteral vaccines which have been thought to confer only moderate and short-term immunity. OBJECTIVES: The objective of this review was to assess the effect of cholera vaccines in preventing cases of cholera and preventing deaths. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, Medline, Embase and reference lists of articles. We handsearched the journal Vaccine, contacted researchers in the field and manufacturers. SELECTION CRITERIA: Randomised and quasi-randomised studies comparing cholera vaccines (killed or live) with placebo, control vaccines or no intervention, or comparing types, doses or schedules of cholera vaccine. We included adults and children irrespective of immune status or special risk category. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of trial quality was done independently by two reviewers. MAIN RESULTS: Thirty-two trials were included. Seventeen efficacy trials of relatively good quality, testing parenteral and oral killed whole cell vaccines and involving over 2. 6 million adults, children and infants were included. Nineteen safety trials have been conducted for both types of killed whole cell vaccines and for live vaccines and have involved 11,459 people. For all types of vaccines compared to placebo, the relative risk of contracting cholera at 12 months was 0.49, 95% confidence interval 0. 41 to 0.59 (random effects model). This translates to an efficacy of 51%, 95% confidence interval 41% to 59%. Both parenteral and oral administration were relatively efficacious, but significant protection extended into the third year for oral killed whole cell vaccines. Children under 5 were only protected for up to a year, while older children or adults were protected for up to three years. Parenteral killed whole cell vaccines were associated with increased systemic and local adverse effects compared to placebo. Oral killed whole cell vaccines or oral live vaccines were not. REVIEWER'S CONCLUSIONS: Cholera killed whole cell vaccines appear to be relatively effective and safe. Live oral recombinant vaccines appear to be safe, but efficacy data are not available. Protection against cholera appears to persist for up to two years following a single dose of vaccine, and for three to four years with an annual booster. PMID- 10796564 TI - Vaccines for preventing anthrax. AB - BACKGROUND: Anthrax is an acute bacterial skin disease which may be fatal. Three anthrax vaccines are commercially available but their comparative effectiveness and safety is not clear. OBJECTIVES: The objective of this review was to assess the effects of human anthrax vaccines in healthy adults and children. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline, Embase and the reference lists of articles. We handsearched the journal Vaccine and contacted researchers in the field. SELECTION CRITERIA: Randomised and quasi randomised trials comparing anthrax vaccines with placebo, vaccines for other diseases or no intervention. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction was conducted independently by the six authors. MAIN RESULTS: Two trials involving 16,052 people were included. Both trials had methodological limitations. Compared to placebo, vaccination was associated with a reduced risk of contracting anthrax (relative risk 0.16, 95% confidence interval 0.07 to 0.35). Compared to placebo, the killed vaccine was associated with a higher incidence and severity of adverse effects (odds ratio 5.15, 95% confidence interval 2.28 to 11.61). Just over 5% of participants in the vaccine group reported adverse effects. The effectiveness of the vaccine does not appear to be influenced by the route of inoculation. REVIEWER'S CONCLUSIONS: Killed anthrax vaccines appear to be effective in reducing the risk of contracting anthrax with a relatively low rate of adverse effects. Further research should be restricted to testing new vaccines only. PMID- 10796565 TI - Vaccines for preventing plague. AB - BACKGROUND: Plague is endemic in China, Mongolia, Burma, Vietnam, Indonesia, India, large parts of Southern Africa, the United States and South America. There are three types of vaccines (live attenuated, killed and F1 fraction) with varying means of administration. OBJECTIVES: The objective of this review was to assess the effects of vaccines to prevent plague. SEARCH STRATEGY: We searched Medline, Embase, the Cochrane Controlled Trials Register and reference lists of articles. We handsearched the journal 'Vaccine' and contacted experts in the field. SELECTION CRITERIA: Randomised trials comparing live and killed plague vaccines against no intervention, placebo, other plague vaccines or vaccines against other disease (control vaccines). DATA COLLECTION AND ANALYSIS: Three reviewers assessed the eligibility of trials. MAIN RESULTS: No trials were included. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the effectiveness of any plague vaccine, or the relative effectiveness between vaccines and their tolerability. Circumstantial data from observational studies suggest that killed types may be more effective and have fewer adverse effects than attenuated types of vaccine. No evidence appears to exist on the long-term effects of any plague vaccine. PMID- 10796566 TI - Vaccines for preventing tick-borne encephalitis. AB - BACKGROUND: Tick-borne encephalitis is a disease of the central nervous system caused by a virus. Other than the vaccine, there is no treatment for the disease. OBJECTIVES: The objective of this review was to assess the effects of vaccines to prevent tick-borne encephalitis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Vaccine Fields Trials Register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles. We also handsearched the journal Vaccine. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing tick-borne encephalitis vaccines against placebo, control vaccines or comparisons of different doses or schedules of tick-borne encephalitis vaccines. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied inclusion criteria. A panel of six assessors examined trial quality. MAIN RESULTS: Five trials were included. They could not be combined for meta-analysis because of differences in comparisons and outcomes. Four types of tick-borne encephalitis vaccines were used. All the vaccines gave seroconversion rates of over 87%. There were frequent reports of systemic and local adverse effects. REVIEWER'S CONCLUSIONS: Tick-borne encephalitis vaccines appear to be highly immunogenic, but the relationship between seroconversion and clinical protection has not been established. Although adverse effects were commonly reported, none were severe or life threatening. PMID- 10796567 TI - Prevention of oral mucositis or oral candidiasis for patients with cancer receiving chemotherapy (excluding head and neck cancer). AB - BACKGROUND: Treatment of cancer with chemotherapy is becoming increasingly more effective but is associated with short and long-term side effects. Oral side effects remain a major source of illness despite the use of a variety of agents to prevent them. OBJECTIVES: To evaluate the effectiveness of oral (and topical) prophylactic agents for oral mucositis and oral candidiasis in patients with cancer (excluding head and neck cancer), compared with placebo or no treatment. SEARCH STRATEGY: Computerised MEDLINE, EMBASE, CINAHL, CANCERLIT, the Cochrane Controlled Trials Register and the Cochrane Oral Health Group Specialist Register search up to July 1999. Reference lists from relevant articles were scanned and the authors of eligible studies were contacted to identify trials and obtain additional information. SELECTION CRITERIA: Studies were selected if they met the following criteria: design - random or quasi-random allocation of participants; participants - anyone with cancer receiving chemotherapy (excluding head and neck cancer); interventions - prophylactic agents prescribed to reduce oral conditions arising from cancer or its treatment; outcomes - mucositis and oral candidiasis. DATA COLLECTION AND ANALYSIS: Information regarding methods, participants, interventions and outcome measures and results were independently extracted, in duplicate, by two reviewers (JC & HW). Specialist advice was sought to categorise interventions. Authors were contacted for details of randomisation and withdrawals and a quality assessment was carried out using the Jadad criteria (Jadad 1998). The Cochrane Oral Health Group statistical guidelines were followed and relative risk values calculated using random effects models where significant heterogeneity was detected (P < 0.1). MAIN RESULTS: Thirty-eight reports of trials were initially included. Two were duplicate reports and nine were excluded as there was no useable information. Of the 27 useable studies 14 had data for mucositis comprising 945 randomised patients and 15 included data for oral candidiasis with 1164 randomised patients. Of the eight prophylactic agents used for mucositis only one, ice chips, was effective (Relative risk 0.57, 95% CI 0.43 to 0.77, chi-square for heterogeneity = 0.26 (df = 1), p = 0.61). The NNT to prevent one extra case of mucositis over the baseline incidence using ice chips was 4 (95%CI: 3 to 7). The NNT for when the baseline incidence of mucositis in the population ranges from 50% to 80% are 5 to 4 respectively. There is evidence that antifungal agents which are partially or fully absorbed from the gastrointestinal tract prevent oral candidiasis and that the partially absorbed agents may be more effective than the fully absorbed agents. The RR for partially absorbed agents was 0.13 (95% CI 0.06 to 0.27, chi-square for heterogeneity = 5.3 (df = 3), P = 0. 15). The NNT to prevent one extra case of oral candidiasis over the baseline incidence using partially absorbed drugs was 3 (95% CI: 3 to 5). The NNT for when the baseline incidence of oral candidiasis in the population ranges from 30% to 70% are 4 to 2 respectively. The general reporting of RCT's was poor however the median Jadad score was acceptable and improved further when the authors provided additional information. The sensitivity analysis confirmed the findings for oral candidiasis. REVIEWER'S CONCLUSIONS: There is some evidence that ice chips prevent mucositis. None of the other prophylactic agents included in this review prevented mucositis. There is evidence that prophylactic use of antifungal agents which are absorbed or partially absorbed from the gastrointestinal tract reduce the clinical signs of oral candidiasis, and the partially absorbed drugs may be more effective. Future trials in this area should address the link between oral and general health including outcomes relevant to the patient. Collaboration between medical and dental teams is indicated. PMID- 10796568 TI - Orthodontic treatment for posterior crossbites. AB - BACKGROUND: 'Posterior crossbite' occurs when the top back teeth bite inside the bottom back teeth. When it affects one side of the mouth the lower jaw may have to move to one side to allow the back teeth to meet together. This movement may have long term effects on the growth of the teeth and jaws. It is unclear what causes posterior crossbites and they may develop or improve at any time from when the baby teeth come into the mouth to when the adult teeth come through. Several treatments have been recommended to correct them. Some treatments widen the upper teeth whilst others are directed at treating the cause of the posterior crossbite e.g. breathing problems or sucking habits. Most treatments have been used at each stage of dental development. OBJECTIVES: The aim of this review was to identify and evaluate orthodontic treatments used to expand the maxillary dentition and / or correct posterior crossbites. SEARCH STRATEGY: All randomised and controlled clinical trials identified from the Cochrane Controlled Trials Register according to the Oral Health Group Search Strategy and stored in the Cochrane Collaboration Oral Health Group Database of Clinical Trials, a MEDLINE search using the Mesh term and free text words, hand searching the British, European and American journals of orthodontics and Angle Orthodontist, and the bibliographies of papers and review articles which reported the outcome of orthodontic treatment to expand the maxillary dentition and/or correct a posterior crossbite that were published as abstracts or papers between 1970 and 1997 in English. SELECTION CRITERIA: All randomised and controlled clinical trials published as full papers or abstracts which reported quantitative data on the outcomes crossbite correction, molar and/or canine expansion, signs and symptoms of temporomandibular joint dysfunction or respiratory disease. DATA COLLECTION AND ANALYSIS: Data were extracted without blinding to the authors, treatments used or results obtained. The first named authors of randomised and controlled clinical trials were written to in an attempt to establish the method of randomisation / allocation and identify unpublished studies. Odds ratio, 95% confidence intervals, relative risk, relative risk reduction, absolute risk reduction, and the number need to treat were calculated for event data. The weighted mean difference and 95% confidence intervals were calculated for continuous data. MAIN RESULTS: Using the search strategy 7 randomised and 5 controlled clinical trials were identified but following correspondence with the authors, 3 of the randomised and 1 of the controlled clinical trials were reclassified giving 5 randomised and 7 controlled clinical trials for inclusion in the review. Trials comparing occlusal grinding in the primary dentition with/without an upper removable expansion appliance in the mixed dentition versus no treatment, banded versus bonded rapid maxillary expansion, banded versus bonded slow maxillary expansion, transpalatal arch with/without buccal root torque and an upper removable expansion appliance versus quad-helix were identified. Occlusal grinding in the primary dentition with/without the addition of an upper removable expansion plate, in the mixed dentition for those children who did not respond to grinding, was shown to be effective in preventing a posterior crossbite in the primary dentition from being perpetuated to the mixed and permanent dentitions. No evidence of a difference in treatment effect (molar and canine expansion) between the test and control intervention was found in the trials which compared banded versus bonded rapid maxillary expansion, banded versus bonded slow maxillary expansion, transpalatal arch with/without buccal root torque, or upper removable expansion appliance versus quad-helix. (ABSTRACT TRUNCATED) PMID- 10796569 TI - Vitamin C for preventing and treating the common cold. AB - BACKGROUND: The role of oral ascorbic acid (vitamin C) in the prevention and treatment of colds remains controversial despite many controlled trials. There have also been a number of efforts to synthesize and/or overview the results of these trials, and controversy over what these overviews tell us. OBJECTIVES: The objective of this review was to answer the following two questions: (1) Does regular high dosage supplementation with vitamin C reduce the incidence of colds? (2) Does taking vitamin C in high doses at the onset of a cold have a therapeutic effect? SEARCH STRATEGY: This review currently deals only with published trials from two previously published reviews by Kleijnen 1989 and Hemila 1992. SELECTION CRITERIA: Randomised and non-randomised trials of vitamin C taken to prevent or treat the common cold. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Thirty trials were included. The quality of the included trials was variable. Vitamin C in doses as high as one gram daily for several winter months, had no consistent beneficial effect on incidence of the common cold. For both preventive and therapeutic trials, there was a consistently beneficial but generally modest therapeutic effect on duration of cold symptoms. This effect was variable, ranging from 0.07% to a 39% reduction in symptom days. The weighted difference across all of the studies revealed a reduction of a little less than half a symptom day per cold episode, representing an 8% to 9% reduction in symptom days. There was no clear indication of the relative benefits of different regimes or vitamin C doses. However in trials that tested vitamin C after cold symptoms occurred, there was some evidence that a large dose produced greater benefits than lower doses. REVIEWER'S CONCLUSIONS: Long term daily supplementation with vitamin C in large doses daily does not appear to prevent colds. There appears to be a modest benefit in reducing duration of cold symptoms from ingestion of relatively high doses of vitamin C. The relation of dose to therapeutic benefit needs further exploration. PMID- 10796570 TI - Improving health professionals' management and the organisation of care for overweight and obese people. AB - BACKGROUND: Obesity is increasing throughout the industrialised world. If left unchecked it will have major implications for both population health and costs to health services. Health professionals have a key role to play in tackling the obesity problem, but little is known about how they may be encouraged to work more effectively with overweight and obese people. OBJECTIVES: The main objective was to determine whether health professionals' management or the organisation of care for overweight and obese people could be improved. SEARCH STRATEGY: We searched the specialised registers of the Cochrane Effective Practice and Organisation of Care Group (May 1997), the Cochrane Depression, Anxiety and Neurosis Group (August 1997), the Cochrane Diabetes Group (August 1997), the Cochrane Controlled Trials Register (September 1997), MEDLINE to January 1998, EMBASE to December 1997, Cinahl (1982 to November 1997), PsycLit (1974 to December 1997), Sigle (1980 to November 1997), Sociofile (1974 to October 1997), Dissertation Abstracts (1861 to January 1998), Conference Papers Index (1973 to January 1998), Resource Database in Continuing Medical Education. We also hand searched seven key journals and contacted experts in the field. SELECTION CRITERIA: Randomised trials, controlled before-and-after studies and interrupted time series analyses of providers' management of obesity or the organisation of care to improve provider practice or patient outcomes. We addressed three a priori comparisons and a fourth post hoc comparison. 1. Interventions aimed at improving health professionals' management or the delivery of health care for overweight/obese patients are more effective than usual care. 2. Interventions aimed at redressing negative attitudes and related practices towards overweight/obese patients are more effective than usual care. 3. Organisational interventions designed to change the structure of services for overweight/obese people are more effective than educational or behavioural interventions for health professionals. 4. Comparisons of different organisational interventions. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Twelve studies were included involving more than 393 providers and 3392 patients. Four studies were identified for comparison 1. Three were professional-oriented interventions (the use of reminders and training) and the fourth was a study of professional and organisational interventions of shared care. No studies were identified for comparisons 2 or 3. Eight studies were identified for post hoc comparison 4. These compared either the deliverer of weight loss interventions or the setting of interventions. The included studies were heterogeneous and of generally poor quality. REVIEWER'S CONCLUSIONS: At present, decisions about improving provision of services must be based on the evidence of patient interventions and good clinical judgement. Further research is needed to identify cost effective strategies for improving the management of obesity. PMID- 10796571 TI - Vitamin E for intermittent claudication. AB - BACKGROUND: It is thought that vitamin E may improve tolerance to intermittent claudication (i.e. pain caused by ischaemia in the muscles of the leg during exercise), thereby relieving the pain, through a variety of mechanisms. OBJECTIVES: The objective of this review was to determine the effects of vitamin E on people with intermittent claudication. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register, reference lists of relevant articles and a library specialising in literature on vitamins. SELECTION CRITERIA: Controlled trials comparing vitamin E with placebo, or other interventions, in patients with intermittent claudication. DATA COLLECTION AND ANALYSIS: Both reviewers extracted data and assessed study quality independently. MAIN RESULTS: Five eligible studies were found with a total of 265, predominantly male, participants. The average age was 57 years. The follow up varied from 12 weeks to 18 months. The trials were small and generally of poor quality. The people studied were reasonably homogeneous but five different doses of vitamin E were used and four different physical outcomes were measured. No trials were identified that compared vitamin E with treatments other than placebo. All trials showed positive effects on one of their main outcomes. No serious adverse effects of vitamin E were reported. Two trials that lasted approximately eight months and used similar doses reported patients' subjective evaluation of the treatment. The relative risk for the combined results of these two trials using a random effects model was 0.57 with a 95% confidence interval of 0.28 to 1.15. REVIEWER'S CONCLUSIONS: While vitamin E - which is inexpensive and has had no serious side effects reported with its use - may have beneficial effects, there is insufficient evidence to determine whether it is an effective treatment for intermittent claudication. PMID- 10796572 TI - Exercise for intermittent claudication. AB - BACKGROUND: Exercise is an inexpensive, low risk option compared with other more invasive therapies for leg pain on walking (intermittent claudication). OBJECTIVES: The objective of this review was to determine the effects of exercise for leg pain. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register, Embase, reference lists of relevant articles, and contacted principal investigators of trials. SELECTION CRITERIA: Randomised trials of exercise regimens in patients with leg pain on walking (intermittent claudication). DATA COLLECTION AND ANALYSIS: At least two reviewers extracted and assessed data trial quality independently. The reviewers contacted investigators to obtain information or data needed for the review that could not be found in published reports. MAIN RESULTS: Fifteen trials were identified that met the inclusion criteria, but five were subsequently excluded because of poor quality. The remaining ten trials involved a total of almost 250 male and female patients with stable leg pain. The follow-up ranged from 12 weeks to 15 months. There was also some variation in the exercise regimens used, although all recommended at least two weekly sessions of, mostly, supervised exercise. All trials used a treadmill walking test as one of the outcome measures. The overall quality of the included trials was generally good, though the trials were all small (20-49 patients). Exercise therapy significantly improved maximal walking time (minutes) (weighted mean difference 6.51, 95% confidence interval 4.36 to 8.66, fixed effect model [FE]), with an overall improvement in walking ability of approximately 150% (range 74% to 230%). Exercise produced significant improvements in walking time compared with both angioplasty at six months (weighted mean difference 3.30, 95% confidence interval 2.21 to 4.39, FE) and antiplatelet therapy (weighted mean difference 1.06, 95% confidence interval 0.15 to 1.97, FE), and did not differ significantly from surgical treatment. In one small trial, exercise was less effective than pentoxifylline (weighted mean difference -0.45, 95% confidence interval -0.66 to -0.24, FE). REVIEWER'S CONCLUSIONS: Exercise is of significant benefit to patients with leg pain. PMID- 10796573 TI - Home care by outreach nursing for chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterised by progressive airflow obstruction, worsening exercise performance and deterioration in health. It is associated with significant morbidity, mortality and costs to health care systems. Care strategies, such as outreach nursing in the community, may reduce this burden. OBJECTIVES: To evaluate the effectiveness of outreach respiratory health care worker programmes for patients with COPD in terms of improving lung function, exercise tolerance and health related quality of life (HRQL) of patient and carer, and reducing mortality and hospital service utilisation. SEARCH STRATEGY: A search was carried out using the Cochrane Airways Group database. Bibliographies of identified RCTs were searched for additional relevant RCTs. Authors of identified RCTS were contacted for other published and unpublished studies. SELECTION CRITERIA: Only randomised control trials of patients with COPD were included. The intervention was an outreach nurse visiting patients in their homes, providing support, education, monitoring patient status and providing liaison with physicians. Interventions that used nurse practitioners who provided therapeutic intervention were also included. Studies in which the therapeutic intervention under test was physical training were not included. DATA COLLECTION AND ANALYSIS: Data extraction and study quality assessment were performed independently by two reviewers. Where further or missing data was required, authors of studies were contacted. MAIN RESULTS: Four studies were found. Three assessed mortality following twelve months of care (n=96, 152 and 301), and one after seven months(n=75). Meta-analysis demonstrated that mortality was not significantly reduced by the intervention, Peto Odds Ratio 0.72; 95 % confidence interval 0.43, 1.21. Post hoc subgroup analysis suggested that mortality was reduced by the outreach nursing programme in patients with less severe disease. Significant improvements in health related quality of life were reported in one study in moderate COPD, but not in a study in patients with severe disease. No changes in lung function or exercise performance were found in the studies where data were available. Hospital admissions were reported in only one study in patients with severe disease and no benefit was observed. REVIEWER'S CONCLUSIONS: Patients with moderate COPD may have mortality and health related quality of life gains from a nursing outreach programme, but there are no data about reductions in hospital utilisation. Patients with severe COPD do not appear to have benefit from such programmes and one large study found no reduction in hospital admissions in such patients. PMID- 10796574 TI - Alexander technique for chronic asthma. AB - BACKGROUND: "The Alexander technique" is a taught form of physical therapy involving a series of movements designed to correct posture and bring the body into natural alignment with the object of helping it to function efficiently, and is reported to aid relaxation. Some practitioners claim benefits for those who desire greater ease and efficiency of breathing, including asthmatics. OBJECTIVES: The objective of this review was to evaluate the efficacy of the Alexander technique in people with chronic, stable asthma. SEARCH STRATEGY: The specialised register of the Cochrane Airways Group, the Cochrane Complementary Medicine Field trials register and the bibliographies of relevant articles were searched. SELECTION CRITERIA: Randomised controlled trials of Alexander technique for the improvement of the symptoms of chronic, stable asthma, comparing the treatment with either another intervention or no intervention. DATA COLLECTION AND ANALYSIS: No trials were found that met the selection criteria. REVIEWER'S CONCLUSIONS: Robust, well-designed randomised controlled trials are required in order to test claims by practitioners that AT can have a positive effect on the symptoms of chronic asthma and thereby help people with asthma to reduce medication. PMID- 10796575 TI - Inhaled steroids for bronchiectasis. AB - BACKGROUND: Bronchiectasis is a progressive condition characterised by irreversible destruction and dilatation of airways, generally associated with chronic bacterial infection. The two distinct therapeutic goals are: symptom control and reduction in morbidity; and prevention of progression of the underlying disease. OBJECTIVES: To determine whether regular inhaled corticosteroids produce improvement in symptom control and whether they beneficially influence the natural history of the disease. SEARCH STRATEGY: The Cochrane Airways Group RCT register and Cochrane Controlled Clinical Trials Register were searched using the following search terms; bronchiectasis AND [corticosteroid* OR beclomethasone OR budesonide OR fluticasone OR triamcinolone OR flunisolide]. Bibliographies of each included RCT was searched for additional trials. Pharmaceutical companies that manufacture inhaled corticosteroids were also contacted. SELECTION CRITERIA: Only randomised double blind studies controlled trials were included. Patients with radiographic evidence of bronchiectasis were included, but patients with cystic fibrosis were excluded. DATA COLLECTION AND ANALYSIS: Data was extracted by one of the reviewers (FR). Continuous outcomes were analysed as effect sizes (weighted mean difference or as standardised mean difference with 95% confidence intervals). MAIN RESULTS: Only two trials on a total of 54 patients could be included. The studies were of 4 and 6 weeks duration. Inhaled corticosteroids had no significant effect on any of the outcomes included in this review, however there was a trend towards improving: FEV1, FVC, PEFR, RV and DLco. REVIEWER'S CONCLUSIONS: In bronchiectasis, regular use of inhaled corticosteroids may improve lung function. The available studies were too short and too small to provide any clear evidence to guide practice. Larger and longer studies should include rate of decline of lung function, exacerbation frequency, hospitalisations and healthy status as outcomes. PMID- 10796576 TI - Corticosteroids for the prevention and treatment of post-extubation stridor in neonates, children and adults. AB - BACKGROUND: Post-extubation stridor may prolong length of stay in the intensive care unit, particularly if airway obstruction is severe and re-intubation proves necessary. Corticosteroids, however, may be associated with adverse effects ranging from hypertension to hyperglycemia, and a more systematic assessment of the efficacy of this therapy is indicated prior to widespread adoption of this practice OBJECTIVES: To determine whether corticosteroids are effective in preventing or treating post-extubation stridor in critically ill infants, children, or adults. SEARCH STRATEGY: Controlled trials were identified through MEDLINE, EMBASE and CINAHL. Bibliographies of all identified trials were examined, and authors of included trials were contacted to confirm the methodology and identify other potentially relevant trials. SELECTION CRITERIA: Any randomized controlled trial that compared administration of corticosteroids by any route with placebo in infants, children, or adults receiving mechanical ventilation via an endotracheal tube in an intensive care unit. DATA COLLECTION AND ANALYSIS: Data from the included studies were extracted independently by two reviewers. The main outcome was the rate of tracheal re-intubation after extubation. The incidence of stridor was examined as a secondary outcome. Subgroup analysis was predetermined to compare preventative use of steroids before extubation and therapeutic use following extubation. Neonates, pediatric patients, and adults were compared separately. A random effects model was used throughout. Methodologic quality of trials was assessed independently by the two reviewers. MAIN RESULTS: Of 251 studies identified, only seven met the criteria for inclusion; three in adults, two in neonates, three in children. All but one examined use of steroids for the prevention of post-extubation stridor; the remaining one concerned treatment of existing post-extubation stridor in children. Patients were drawn from heterogeneous medical/surgical populations. Dexamethasone given intravenously at least once prior to extubation was the most common steroid regimen utilized (uniformly in neonates and children). Prophylactic intervention tended to decrease re-intubation rates among neonates and children, but did not reach statistical significance (neonates RR=0.1, 95% CI 0.01, 1.68; children RR=0.49, 95% CI 0.01, 19.65). Post-extubation stridor was reduced in children (n=216: RR=0.53, 95% CI 0.28, 0.97) but not in neonates. In the neonatal studies, a lower re-intubation rate was seen only in high risk patients treated with multiple doses of steroids around the time of extubation. In three adult studies (total n=1047), no difference in post-extubation stridor (RR=0.86, 95% CI 0.57, 1.30) or re-intubation rates (RR=0.95, 95% CI 0.52, 1.72) was detected. Side effects were reported seldomly and could not be aggregated. REVIEWER'S CONCLUSIONS: In neonates, there is a trend towards a reduced incidence of re-intubation in neonates receiving prophylactic dexamethasone prior to extubation. In children, prophylactic administration of dexamethasone prior to elective extubation reduces the incidence of post-extubation stridor, but the evidence is insufficient to conclude that rates of re-intubation are reduced. In adults, corticosteroids do not appear to reduce the need for re-intubation. PMID- 10796577 TI - Lung volume reduction surgery for diffuse emphysema. AB - BACKGROUND: Lung volume reduction surgery (LVRS) has been re-introduced for treating patients with severe diffuse emphysema. OBJECTIVES: To assemble evidence from randomised controlled trials for the effectiveness of LVRS, and identify optimal surgical techniques, those patients who benefit most and those for whom it should be avoided. SEARCH STRATEGY: Randomised controlled trials were identified using the Cochrane Airways Group COPD register using the terms: emphysema AND (emphysema surgery OR lung volume reduction surgery OR LVRS OR volume reduction surgery OR pneumectomy OR reduction pneumoplasty OR lung reduction surgery). The Cochrane Controlled Clinical Trials Register was also searched using these terms. SELECTION CRITERIA: Randomised controlled trials that studied the safety and efficacy of LVRS in patients with diffuse emphysema were included. Studies were excluded if they investigated giant or bullous emphysema. DATA COLLECTION AND ANALYSIS: Two independent reviewers assessed trials for inclusion and extracted data. MAIN RESULTS: Only one randomised trial of LVRS for diffuse emphysema was identified. This compared stapled unilateral thoracoscopic lung reduction coupled with bovine pericardium reinforcement with a unilateral neodymium:yttrium aluminium garnet laser contact reduction. A total of 72 patients were studied. Both arms included post-operative rehabilitation and appeared to be well matched at randomisation. Improvement in FEV1 & FVC at six months was significantly greater in the staple treated group (p < 0.01 & p < 0. 07 respectively), but absolute increases were small. Need for supplemental oxygen was reduced significantly more in the staple treated group; Peto Odds Ratio (OR) 4.05; 95% confidence interval (CI) 1.40, 11.71. Quality of life improved more in the staple treated group (OR 5.36; 95% CI 2.13,13.47). The rate of delayed pneumothorax in the laser treated group was significantly higher (OR 10.46; 95% CI 1.98, 55.30). REVIEWER'S CONCLUSIONS: There is no randomised controlled trial evidence concerning the efficacy of LVRS for diffuse emphysema compared to optimal conservative medical therapy. Stapling is more effective than laser resection and has a lower complication rate. LVRS should not be applied routinely until results of large trials currently underway become available. PMID- 10796578 TI - Manual therapy for asthma. AB - BACKGROUND: A variety of manual therapies with similar postulated biologic mechanisms of action are commonly used to treat patients with asthma. Manual therapy practitioners are also varied, including physiotherapists, respiratory therapists, chiropractic and osteopathic physicians. A systematic review across disciplines is warranted. OBJECTIVES: To evaluate the evidence for the effects of manual therapies for treatment of patients with bronchial asthma. SEARCH STRATEGY: Trials were searched in computerized general (EMBASE, CINAHL and MEDLINE) and specialized databases (Cochrane Complementary Medicine Field, Cochrane Rehabilitation Field, ICL, and MANTIS). In addition, bibliographies from included studies were assessed, and authors of known studies were contacted for additional information about published and unpublished trials. Date of most recent search: December 1998. SELECTION CRITERIA: Trials were included if they: (1) were randomised; (2) included asthmatic children or adults; (3) examined one or more types of manual therapy; and (4) included clinical outcomes. DATA COLLECTION AND ANALYSIS: All three reviewers independently extracted data and assessed trial quality using a standard form. MAIN RESULTS: From an initial 316 unique citations, 48 full text articles were retrieved and evaluated, which resulted in nine citations to five RCTs (290 patients) suitable for inclusion. Trials could not be pooled statistically because studies that addressed similar interventions used disparate patient groups or outcomes. The methodological quality of one of two trials examining chiropractic manipulation was good and neither trial found significant differences between chiropractic spinal manipulation and a sham manoeuvre on any of the outcomes measured. Quality of the remaining three trials was poor. One small trial compared massage therapy with a relaxation control group and found significant differences in many of the lung function measures obtained. However, this trial had poor reporting characteristics and the data have yet to be confirmed. One small trial compared chest physiotherapy to placebo and one small trial compared footzone therapy to a no treatment control. Neither trial found differences in lung function between groups. REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the use of manual therapies for patients with asthma. There is a need to conduct adequately-sized RCTs that examine the effects of manual therapies on clinically relevant outcomes. Future trials should maintain observer blinding for outcome assessments, and report on the costs of care and adverse events. Currently, there is insufficient evidence to support or refute the use of manual therapy for patients with asthma. PMID- 10796579 TI - Surgery for obstructive sleep apnoea. AB - BACKGROUND: Obstructive sleep apnoea is the periodic reduction (hypopnoea) or cessation (apnoea) of airflow during sleep. The syndrome is associated with loud snoring, disrupted sleep and observed apnoeas. Beside conservative treatments there are a range of 8 broad categories of operative treatments for this condition. Surgical treatments aim to relieve the obstruction by either increasing the surface area OBJECTIVES: Surgery for obstructive sleep apnoea aims to relieve the obstruction by increasing the surface area of the airway, bypassing the pharyngeal airways or removing a lesion. The objective of this review was to assess the effects of any type of surgery for the treatment of obstructive sleep apnoea. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. We contacted experts in the field, research dissemination bodies and other Cochrane Review Groups. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing any surgical intervention for obstructive sleep apnoea with other surgical or non-surgical interventions for obstructive sleep apnoea or no intervention. DATA COLLECTION AND ANALYSIS: No completed trials were identified. MAIN RESULTS: No studies fulfilled the inclusion criteria. REVIEWER'S CONCLUSIONS: There is an urgent need for high quality randomised controlled trials to be carried out in the field of surgery for obstructive sleep apnoea. More research should also be undertaken to identify and standardise techniques to determine the site of airways obstructions. PMID- 10796580 TI - Limited (information only) patient education programs for adults with asthma. AB - BACKGROUND: A key component of many asthma management guidelines is the recommendation for patient education and regular medical review. A number of controlled trials have been conducted to measure the effectiveness of asthma education programmes. These programmes improve patient knowledge, but their impact on health outcomes is less well established. At its simplest level, education is limited to the transfer of information about asthma, its causes and its treatment. This review focused on the effects of limited asthma education. OBJECTIVES: The objective of this review was to assess the effects of limited (i.e. information only) asthma education on health outcomes in adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised and controlled trials of individual asthma education involving information transfer only in adults over 16 years of age. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for missing information. MAIN RESULTS: Eleven trials were included. They were of variable quality. Limited asthma education did not reduce hospitalisation for asthma (weighted mean difference -0.03 average hospitalisations per person per year, 95% confidence interval -0.09 to 0.03). There was no effect on doctor visits, lung function and medication use. The effects on asthma symptoms were variable. There was no reduction in days lost from normal activity, but perceived asthma symptoms did improve after limited asthma education (odds ratio 0.40, 95% confidence interval 0.18 to 0.86). In one study, limited asthma education was associated with reduced emergency department visits (weighted mean difference -2.76 average visits per person per year, 95% confidence interval -4.34 to 1.18). REVIEWER'S CONCLUSIONS: Use of limited asthma education as it has been practiced does not appear to improve health outcomes in adults with asthma. However the use of information in the emergency department may be effective, but this needs to be confirmed. PMID- 10796581 TI - Mass media interventions for preventing smoking in young people. AB - BACKGROUND: The mass media have been used as a way of delivering preventive health messages. They have the potential to reach and to modify the knowledge, attitudes and behaviour of a large proportion of the community. OBJECTIVES: To determine the effectiveness of mass media campaigns in preventing the uptake of smoking in young people. SEARCH STRATEGY: We searched Medline, and 28 other electronic databases. Handsearching of key journals was also carried out, the bibliographies of identified studies were checked for additional references and contact with content area specialists was made. Date of last search June 1998. SELECTION CRITERIA: Randomised trials, controlled trials without randomisation and time series studies that assessed the effectiveness of mass media campaigns (defined as channels of communication such as television, radio, newspapers, bill boards, posters, leaflets or booklets intended to reach large numbers of people and which are not dependent on person to person contact) in influencing the smoking behaviour (either objective or self-reported) of young people under the age of 25 years. DATA COLLECTION AND ANALYSIS: Information relating to the characteristics and the content of media interventions, participants, outcomes and methods of the study was abstracted by one reviewer and checked by a second. Studies were combined using qualitative narrative synthesis. MAIN RESULTS: Six out of a total of 63 studies reporting information about mass media smoking campaigns met all of the inclusion criteria. All six studies used a controlled trial design. Two studies concluded that the mass media were effective in influencing the smoking behaviour of young people. Both of the effective campaigns had a solid theoretical basis, used formative research in designing the campaign messages and message broadcast was of reasonable intensity over extensive periods of time. REVIEWER'S CONCLUSIONS: There is some evidence that the mass media can be effective in preventing the uptake of smoking in young people, but overall the evidence is not strong. PMID- 10796582 TI - Group behaviour therapy programmes for smoking cessation. AB - BACKGROUND: Group therapy offers individuals the opportunity to learn behavioural techniques for smoking cessation, and to provide each other with mutual support. OBJECTIVES: The aims of this review were to determine the effects of smoking cessation programmes delivered in a group format compared to self-help materials, or to no intervention; to compare the effectiveness of group therapy and individual counselling; to determine the effect of adding group therapy to advice from a health professional or nicotine replacement and to determine the rate at which offers of group therapy are taken up. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: We considered randomised trials which compared group therapy with self-help, individual counselling, another intervention or usual care or waiting list control. We also considered trials which compared two group programmes with manipulation of the group interaction and social support components. We included those trials with a minimum of two group meetings, and follow-up of smoking status at least six months after the start of the programme. We excluded trials in which group therapy was provided to both active therapy and placebo arms of trials of pharmacotherapies, unless they had a factorial design. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the nature of the groups and the controls, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Subjects lost to follow-up were counted as smokers. Where possible, we performed meta-analysis using a fixed effects model. MAIN RESULTS: Thirteen studies compared a group programme with a self-help programme. There was an increase in cessation with the use of a group programme (odds ratio 2.10, 95% confidence interval 1.64 to 2.70). Group programmes were more effective than no intervention or minimal contact interventions (odds ratio 1.91, 95% confidence interval 1.20 to 3. 04). There was no evidence from two trials that group therapy was more effective than a similar intensity of individual counselling. There was no evidence that manipulating the social interactions between participants in a group programme had an effect on outcome. There was limited evidence that the addition of group therapy to other forms of treatment, including advice from a health professional or nicotine replacement produced extra benefit. There was variation in the extent to which those offered group therapy accepted the treatment. REVIEWER'S CONCLUSIONS: There is evidence that groups are better than self-help, and other less intensive interventions. There is not enough evidence on their effectiveness compared to intensive individual counselling. PMID- 10796583 TI - Hypnotherapy for smoking cessation. AB - BACKGROUND: Hypnotherapy is widely promoted as a method for aiding smoking cessation. It is proposed to act on underlying impulses to weaken the desire to smoke or strengthen the will to stop. OBJECTIVES: The objective of this review was to evaluate the effects of hypnotherapy for smoking cessation. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: We considered randomised trials of hypnotherapy which reported smoking cessation rates at least six months after the beginning of treatment. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data on the type of subjects, the type and duration of the hypnotherapy, the nature of the control group,the outcome measures, method of randomisation, and completeness of follow up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Subjects lost to follow-up were counted as smokers. Where possible, we performed meta-analysis using a fixed effects model. MAIN RESULTS: Nine studies compared hypnotherapy with 14 different control interventions. There was significant heterogeneity between the results of the individual studies, with conflicting results for the effectiveness of hypnotherapy compared to no treatment or to advice. We therefore did not attempt to calculate pooled odds ratios for the overall effect of hypnotherapy. There was no evidence of an effect of hypnotherapy compared to rapid smoking or psychological treatment. REVIEWER'S CONCLUSIONS: We have not shown that hypnotherapy has a greater effect on six month quit rates than other interventions or no treatment. The effects of hypnotherapy on smoking cessation claimed by uncontrolled studies were not confirmed by analysis of randomised controlled trials. PMID- 10796584 TI - Mecamylamine (a nicotine antagonist) for smoking cessation. AB - BACKGROUND: Mecamylamine is a nicotine antagonist (that is it blocks the effect of nicotine). The rationale for its use in smoking cessation is that it may block the rewarding effect of nicotine and thus reduce the urge to smoke. OBJECTIVES: The objective of this review was to determine the effectiveness of mecamylamine in promoting smoking cessation, either alone or in combination with nicotine replacement therapy. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. SELECTION CRITERIA: Randomised trials of mecamylamine, either alone or in combination with nicotine replacement therapy, which reported smoking cessation rates at least six months after intervention. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the dose and duration of the mecamylamine and nicotine treatments, side-effects of treatment, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was sustained abstinence from smoking (biochemically validated) after at least six months follow-up in patients smoking at baseline. Smokers lost to follow-up were regarded as being continuing smokers. Because of the preliminary nature of available data, we did not perform meta analysis but report the results narratively. MAIN RESULTS: We identified two studies, both from the same investigators. In a study of 48 volunteers, a combination of mecamylamine plus nicotine patch was more effective than nicotine patch alone (abstinence rate at one year 37.5% vs 4.2%). In a second study, 80 volunteers were treated for four weeks prior to cessation with one of four treatments: 1. Nicotine patch plus mecamylamine capsules 2. Nicotine alone 3. Mecamylamine alone 4. No active drug. All four groups received combination treatment with nicotine and mecamylamine after the scheduled quit date. The abstinence rates in these four groups were respectively 40%, 20%, 15% and 15%. The higher abstinence rate in the group treated with combination therapy was not statistically significant. The authors reported a statistically significant benefit of mecamylamine using Kaplan-Meier survival analysis. In the doses used, mecamylamine was well tolerated, although up to 40% of subjects required reductions in dose, usually because of constipation. REVIEWER'S CONCLUSIONS: Data from two small studies suggest that the combination of nicotine and mecamylamine may be superior to nicotine alone in promoting smoking cessation. However, these results require confirmation in larger studies before the treatment can be recommended clinically. PMID- 10796585 TI - Piracetam for dementia or cognitive impairment. AB - OBJECTIVES: To determine the clinical efficacy of piracetam for the features of dementia or cognitive impairment, classified according to the major subtypes of dementia: vascular, Alzheimer's disease or mixed vascular and Alzheimer's disease or unclassified dementia or cognitive impairment not fulfilling the criteria for dementia. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials was searched using the terms "piracetam", "nootropic" and "2-oxo-l-pyrrolidine acetamide". Electronic bibliographic databases including Medline, Embase, PychLit, Current Contents, Sociofile were searched back to 1966 with the terms piracetam, nootropics, 2-oxo-1-pyrrolidine and trials. In addition the pharmaceutical company responsible for marketing most of the piracetam worldwide, UCB Pharma, provided a comprehensive list of abstracts, which included many unpublished studies. As many of these unpublished, placebo control studies will be reviewed as possible. SELECTION CRITERIA: All unconfounded trials specified as randomised in which treatment with piracetam was administered for more than a day and compared with placebo in patients with dementia of the Alzheimer's type, vascular dementia or mixed vascular and Alzheimer's disease or uncalssified dementia or cognitive impairment not fulfilling the criteria for dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Each study was independently verified as fulfilling the inclusion criteria. Studies were rated for methodological quality by assessment of blinding and loss before analysis as described by Jadad et al. (1996). Studies were pooled if appropriate and possible, and the pooled odds ratios (95%CI) or the average differences (95%CI) were estimated. Where possible, intention-to-treat data were used. Sensitivity analyses were performed to determine if successive elimination of those studies performing most poorly on these quality criteria changed the effect estimate. MAIN RESULTS: Unfortunately, many of these studies were crossover in design and data were unavailable from the first period. In many other studies data were not able to be extracted from the first period. From the data that were pooled there was only one outcome where significant amounts of evidence were available, Global Impression of Change. There was evidence of heterogeneity in the results from the individual studies, Chi squared test = 20.8 (df=5). Using a fixed effects model the odds ratio for improvement in the Piracetem group compared with the Placebo group was 3.55, [95% CI][2.45, 5.16]. If a random effects model was used the odds ratio was 3.47 [1.29, 9.30]. If one single-blind study was excluded, the fixed effects model yielded an odds ratio of 3.36 [2.29, 4.99] and if a random effects model was applied then the odds ratio was 2.89 [1.01, 8.24]. The evidence of effects on cognition and other measures, was inconclusive. REVIEWER'S CONCLUSIONS: At this stage the evidence available from the published literature does not support the use of Piracetem in the treatment of people with dementia or cognitive impairment because effects were found only on global impression of change but not on any of the more specific measures. There is a need for further evaluation of piracetam by : 1) Obtaining the data from these studies for an individual patient database review, 2) Performing a randomised trial of Piracetam in patients with diagnoses made by currently accepted diagnostic criteria. Piracetam should be trialled for a period of at least 6 months and preferably longer. Specific cognitive instruments which are sensitive to change, Clinician Global Impression of Change, levels of dependency and caregiver quality of life scales should also be incorporated in such a study. PMID- 10796586 TI - Lecithin for dementia and cognitive impairment. AB - BACKGROUND: People with Alzheimer's disease have been found to have a relative lack of the enzyme responsible for converting choline into acetylcholine within the brain. Lecithin is a major dietary source of choline, so extra consumption may assist in the production of acetylcholine and reduce some of the symptoms of dementia. OBJECTIVES: To determine the efficacy of lecithin in the treatment of dementia or cognitive impairment. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials has been searched, as have the electronic databases MEDLINE, EMBASE, Psychlit, ISI and Current Contents. Reference lists and relevant books have been examined. SELECTION CRITERIA: All unconfounded, randomised trials comparing lecithin with placebo in a treatment period longer than one day, in patients with dementia of the Alzheimer type, vascular dementia, mixed vascular and Alzheimer's disease, unclassified or other dementia or unclassified cognitive impairment not fulfilling the criteria for dementia are eligible for inclusion. DATA COLLECTION AND ANALYSIS: Data are extracted by two independent reviewers and cross checked. Meta-analyses are performed when more than one trial provide data on a comparable outcome on sufficiently similar patients. Random effects analyses are performed whenever heterogeneity between results appears to be present. Standardised mean difference are used due do the use of different scales and periods of treatment. Odds ratios for dichotomous data are pooled using the Mantel-Haenszel or DerSimonian and Laird methods. MAIN RESULTS: Eleven randomised trials have been identified involving patients with Alzheimer's disease (265 patients) and Parkinsonian dementia (21 patients). No trials reported any clear clinical benefit of lecithin. Few trials contributed data to meta-analyses. The only statistically significant result was in favour of placebo for adverse events, based on one trial, which appears likely to be a spurious result. REVIEWER'S CONCLUSIONS: Evidence from randomised trials does not support the use of lecithin in the treatment of patients with dementia or cognitive impairment. A moderate effect cannot be ruled out, but results from the small trials to date do not indicate priority for a large randomised trial. PMID- 10796587 TI - Cyclical progestogens for heavy menstrual bleeding. AB - BACKGROUND: Excessively heavy menstrual bleeding (HMB) or menorrhagia is an important cause of ill health in women. Eighty per cent of women treated for HMB have no anatomical pathology and so medical therapy, with the avoidance of possibly unnecessary surgery, is an attractive alternative. Of the wide variety of medications used to reduce heavy menstrual bleeding, oral progestogens are the most commonly prescribed in many western countries, although there is little objective evidence to support their use, especially in women with ovulatory menstruation. This review assesses the effectiveness of 2 different regimens of oral progestogens in reducing ovulatory HMB. OBJECTIVES: The primary objective of this review is to investigate the effectiveness of oral progestogen therapy taken either during the luteal phase or for a longer course of 21 days in achieving a reduction in menstrual blood loss in women of reproductive years with heavy menstrual bleeding (HMB). SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Register of Trials, MEDLINE, EMBASE, PsychLIT, Current Contents, Biological Abstracts, Social Sciences Index and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles. In most cases, the first author of each included trial was contacted. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of oral progestogen therapy versus placebo or other medical treatments in women of reproductive years with regular heavy periods measured either objectively or subjectively and with no pathological or iatrogenic causes for their heavy menstrual blood loss. DATA COLLECTION AND ANALYSIS: Seven randomised controlled trials (RCTs) were identified that fulfilled the inclusion criteria for this review. The reviewers extracted the data independently and odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data. MAIN RESULTS: No RCTs comparing progestogen treatment with placebo were identified. Comparisons between oral progestogens and other medical therapies were assessed separately according to dosage regimen, progestogens given during the luteal phase of the menstrual cycle and progestogens given for 21 days between day 5 and 26. Progestogen therapy during the luteal phase was significantly less effective at reducing menstrual blood loss when compared with tranexamic acid, danazol and the progesterone releasing intrauterine system (IUS) and there was also a strong non-significant trend in favour of nonsteroidal anti inflammatory drugs (NSAIDs). Duration of menstruation was significantly longer with the progesterone IUS when compared with oral progestogen therapy but significantly shorter under danazol treatment. Compliance and acceptability of treatment where measured did not differ between treatments. Adverse events were significantly more likely under danazol when compared with progestogen treatment. Change in quality of life was not significantly different with progestogen and tranexamic acid therapy but there was a non-significant trend in favour of tranexamic acid for all three categories. Progestogen therapy administered from day 5 to 26 of the menstrual cycle was significantly less effective at reducing menstrual blood loss than the progestogen releasing intrauterine system (LNG IUS) although the reduction from baseline was significant for both groups. The odds of the menstrual period becoming "normal" (ie <80mls/cycle) were also less likely in patients treated with norethisterone (NET) (days 5 to 26) compared to patients treated with LNG IUS. A significantly higher proportion of NET patients found their treatment unacceptable compared to LNG IUS patients. However, the adverse events breast tenderness and intermenstrual bleeding were more likely in the patients with the IUS. (ABSTRACT TRUNCATED) PMID- 10796588 TI - Treatment programmes for people with both severe mental illness and substance misuse. AB - BACKGROUND: Substance misuse in the context of severe mental illness can have detrimental effects. A variety of treatments exist, but the drive has been to provide programmes integrating treatment of both substance misuse and severe mental illness. Such programmes require additional resources and may require radical redesign of service delivery systems. OBJECTIVES: To evaluate the effectiveness of treatment programmes within psychiatric care for people with problems of both substance misuse and serious mental illness. SEARCH STRATEGY: Biological Abstracts (1985-1998), CINAHL (1982-1998), The Cochrane Library (Issue 3, 1998), The Cochrane Schizophrenia Group's Register of trials (August 1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1974-1998) and Sociofile (1974 1998) were comprehensively searched. Citations of all trials were searched and further studies sought from published trials and their authors. SELECTION CRITERIA: All randomised trials of any programme of substance misuse treatment for people with serious mental illness and current problems of substance misuse. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were also independently extracted. For homogeneous dichotomous data the Peto odds ratio (OR), and 95% confidence intervals (CI) were calculated on an intention-to-treat basis. MAIN RESULTS: Six relevant studies, four of which were small, were identified. In general, the quality of design and reporting was not high. Clinically important outcomes such as relapse of severe mental illness, violence to others, patient or carer satisfaction, social functioning and employment were not reported. There is no clear evidence supporting an advantage of any type of substance misuse programme for those with serious mental illness over the value of standard care. No one programme is clearly superior to another. REVIEWER'S CONCLUSIONS: The problems posed by substance misuse in the context of severe mental illness will not go away. The current momentum for integrated programmes is not based on good evidence. Implementation of new specialist substance misuse services for those with serious mental illnesses should be within the context of simple, well designed controlled clinical trials. PMID- 10796589 TI - Intravenous immunoglobulin for treating sepsis and septic shock. AB - OBJECTIVES: Death from severe sepsis and septic shock is common, and researchers have explored whether antibodies to the endotoxins in some bacteria reduces mortality. This review summarises the effects of intravenous immunoglobulin (IVIG) in patients with bacterial sepsis or septic shock on mortality, bacteriological failure rates, and duration of stay in hospital. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline 1966 to April 1999, EMBASE 1988 to February 1999; we contacted investigators active in the field for unpublished data. SELECTION CRITERIA: Randomised trials comparing intravenous immunoglobulin (monoclonal or polyclonal) with placebo or no intervention, in patients with bacterial sepsis or septic shock. DATA COLLECTION AND ANALYSIS: Inclusion criteria, trial quality assessment, and data abstraction were done in duplicate. We conducted pre-specified subgroup analyses by type of immunoglobulin preparation. MAIN RESULTS: Twenty-three out of 49 studies met our inclusion criteria. Overall mortality was reduced in patients who received polyclonal IVIG (n=413; RR=0.60; 95% CI 0.47 to 0.76). Mortality was not reduced among patients who received monoclonal antibodies such as anti-endotoxins (n=1,736 in 4 good quality studies; RR=0.98; 95% CI 0.86 to 1.12) or anti-cytokines (n=4,318; RR=0.93; 95% CI 0.86 to 1.01). A few studies measured secondary outcomes (deaths from sepsis or length of hospitalisation) but no differences in the intervention and control groups were identified except among those who received polyclonal IVIG, where sepsis-related mortality was significantly reduced (n=161; RR=0.35; 95% CI 0.18 to 0.69). REVIEWER'S CONCLUSIONS: In our opinion, polyclonal IVIG significantly reduces mortality and can be used as an adjuvant treatment for sepsis and septic shock. Adjunctive therapy with monoclonal IVIGs remains experimental. PMID- 10796590 TI - Polysaccharide vaccines for preventing serogroup A meningococcal meningitis. AB - BACKGROUND: Controlled trials over two decades ago showed that the polysaccharide vaccine prevented meningococcal meningitis. Subsequent observational studies suggested variations in the level and duration of protection, particularly among young children. OBJECTIVES: To determine the effect of polysaccharide serogroup A vaccine for preventing serogroup A meningococcal meningitis. SEARCH STRATEGY: MEDLINE and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised and other prospective controlled trials. DATA COLLECTION AND ANALYSIS: One reviewer collected data and assessed the methodologic quality of the trials. Data were pooled using the Exact method to assess vaccine efficacy at one, two and three years post-vaccination. MAIN RESULTS: The protective effect within the first year was consistent across all eight trials, including one with participants exclusively under six years of age (in Finland); overall vaccine efficacy was 95% (Exact 95% CI 87%, 99%). The Finnish trial lacked statistical power to assess the effect of a booster dose given to children less than 18 months old. In the three other trials that included children less than six years old (one in Sudan and two in Nigeria), none of the vaccinated children developed meningitis, but the statistical significance of this finding was undetermined. Protection extended into the second (in two studies) and third (in one study) years after vaccination, but these results were also not statistically significant. REVIEWER'S CONCLUSIONS: When compared with current recommendations, the methodological quality and relative incompleteness of the published reports could arguably render the trials invalid for this review. However, it was unlikely that the results of the trials in such diverse settings would have been biased towards a strong and consistent protective effect. Immunogenicity trials were not included in this review. Stage two of the review will assess the vaccine effect from observational studies. PMID- 10796592 TI - Surgery for deep venous incompetence. AB - BACKGROUND: Chronic deep venous incompetence (DVI) is a troublesome condition with a range of symptoms in the legs including recurrent ulcers, pain and swelling. It is caused by incompetent vein valves and/or the blockage of large calibre leg veins. OBJECTIVES: To establish the effectiveness of various surgical procedures for treating DVI. SEARCH STRATEGY: Trials were identified from the Cochrane Peripheral Vascular Diseases Group's Specialised Trials Register, reference lists of relevant studies, and through contact with principal investigators of identified trials and world experts in deep venous surgery. SELECTION CRITERIA: Randomised controlled trials of surgical treatment for patients with DVI. Trials were selected by AA and checked by SCH. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently. A variety of outcome measures were reported including ambulatory venous pressure (AVP) and venous refill time (VRT). MAIN RESULTS: Only one trial met the inclusion criteria, none was excluded. The trial compared external valvuloplasty using limited anterior plication (LAP) in combination with ligature of incompetent superficial veins (ligation and LAP) against ligation only. The trial participants had primary valvular incompetence with mild to moderate symptoms but no venous ulcers. Ligation and LAP produced significant improvement in AVP: the mean difference between the Ligation and LAP group and the Ligation only group was -15 torr (weighted mean difference [WMD] -20.9, -9.0, confidence interval [CI] 95% fixed) at one year and -15 torr (WMD -21, -8.9, 95% CI fixed) at two years. However, there was no statistically significant improvement in VRT, the mean difference between the groups at one year was 2 seconds (WMD -2.7, 6.7; 95% CI fixed) and at two years was 4 seconds (WMD -0.7, 8.7; 95% CI fixed). AVP values after surgery remained relatively high. Nine out of eleven valves repaired remained competent after two years of follow up. No complications occurred. The overall mean score for clinical outcome was +2 (moderate improvement) in the Ligation and LAP group. This compared with +1 (mild improvement) in the Ligation only group. REVIEWER'S CONCLUSIONS: The results of one small trial showed that ligation and LAP produced a moderate improvement for two years after surgery, in patients with mild to moderate DVI caused by primary valvular incompetence. However, there is not sufficient evidence to recommend the treatment to this subgroup of patients with DVI. PMID- 10796591 TI - Short course antibiotics for acute otitis media. AB - BACKGROUND: Otitis media is a common pediatric problem, for which antibiotics are frequently prescribed. OBJECTIVES: To determine the effectiveness of a short course of antibiotics (less than seven days) in comparison to a longer course (seven days or greater) for the treatment of acute otitis media in children. SEARCH STRATEGY: The medical literature was searched for randomized controlled studies of the treatment of ear infections in children with antibiotics published from January 1966 to July 1997. Search last updated March 1998. SELECTION CRITERIA: Studies were included if they met the following criteria: subjects one month to 18 years of age, clinical diagnosis of ear infection, no previous antimicrobial therapy and randomization to treatment with less than seven days versus seven days or more of antibiotics. DATA COLLECTION AND ANALYSIS: Data on treatment outcomes were extracted from individual studies, and combined in the form of a summary odds ratio. A summary odds ratio (OR) equivalent to one indicated that the treatment failure rate following less than seven days of antibiotic treatment was similar to the failure rate following seven days or more of antibiotic. MAIN RESULTS: The summary OR for treatment outcomes at eight to 19 days in 1,524 children treated with short-acting antibiotics for five days versus eight to 10 days was 1.52, 95% CI: 1.17-1.98, but by 20 to 30 days outcomes between treatment groups (n=2,115) were comparable (OR=1.22, 95% CI:0.98-1.54). The absolute difference in treatment failure (Random effects model RD=2.9%, 95%CI:-0.3% to 6.1%) at 20 to 30 days suggests that at minimum 17 children would need to be treated with the long course of short-acting antibiotics to avoid one treatment failure. Similarity in outcomes was observed for up to three months following therapy (OR=1.16,95% CI=0.9-1.5). Comparable outcomes were shown between treatment with ceftriaxone or azithromycin, and more than seven days of other antibiotics. REVIEWER'S CONCLUSIONS: This review suggests that five days of short-acting antibiotic is effective treatment for uncomplicated ear infections in children. PMID- 10796593 TI - Fixed dose subcutaneous low molecular weight heparins versus adjusted dose unfractionated heparin for venous thromboembolism. AB - BACKGROUND: Low molecular weight heparins have been shown to be effective and safe for prevention of venous thromboembolism. There is accumulating evidence that these new anticoagulants are also effective and safe for treatment of venous thromboembolism. OBJECTIVES: The objective of this review was to determine the effect of fixed-dose, subcutaneous low molecular weight heparins compared with adjusted-dose, intravenous or subcutaneous, unfractionated heparin for initial treatment of acute deep venous thrombosis or pulmonary embolism. SEARCH STRATEGY: Trials were identified from the Cochrane Peripheral Vascular Diseases Group trials register and LILACS. The reviewers contacted colleagues and representatives of pharmaceutical companies for additional information about trials. SELECTION CRITERIA: Randomised trials comparing fixed-dose, subcutaneous low molecular weight heparin with adjusted-dose, intravenous or subcutaneous, unfractionated heparin in patients with venous thromboembolism. DATA COLLECTION AND ANALYSIS: Two reviewers assessed trials for inclusion and quality, and extracted data independently. MAIN RESULTS: Fourteen studies with a total of 4754 patients were included. By the end of follow up in ten trials, thrombotic complications occurred in 86 (4.3%) of the 1998 patients treated with low molecular weight heparin, compared with 113 (5.6%) of the 2021 patients treated with unfractionated heparin (odds ratio 0.76, 95% confidence interval 0.57 to 1.01). In eight trials a reduction in thrombus size was shown by 60% treated with low molecular weight heparin and 54% treated with unfractionated heparin (odds ratio 0.77, 95% confidence interval 0.61 to 0.97). At the end of the initial treatment period, in all 14 of the trials, major haemorrhages occurred in 30 (1.3%) of the 2353 patients treated with low molecular weight heparin, compared with 51 (2.1%) of the 2401 patients treated with unfractionated heparin (odds ratio 0.60, 95% confidence interval 0.39 to 0.93). By the end of follow up in 11 trials, 135 (6.4%) of the 2108 patients treated with low molecular weight heparin had died, compared with 172 (8.0%) of the 2137 patients treated with unfractionated heparin (odds ratio 0.78, 95% confidence interval 0.62 to 0.99). Five studies with a total of 1636 patients examined proximal (above the knee) thrombosis; 814 treated with low molecular weight heparin and 822 with unfractionated heparin. A sub-analysis of these trials showed statistically significant reductions favouring the action of low molecular weight heparin in three areas: thrombotic complications; major haemorrhages; and overall mortality. By the end of follow up 39 (4. 8%) patients treated with low molecular weight heparin had thrombotic complications, compared with 64 (7.8%) treated with unfractionated heparin (odds ratio 0.60, 95% confidence interval 0. 40 to 0.89). Major haemorrhages occurred in 8 (1.0%) treated with low molecular weight heparin, compared with 68 (8.3%) treated with unfractionated heparin (odds ratio 0.44, 95% confidence interval 0. 21 to 0.95). By the end of follow up, 44 (5.4%) treated with low molecular weight heparin had died, compared with 68 (8.3%) treated with unfractionated heparin (odds ratio 0.64, 95% confidence interval 0.43 to 0.93). REVIEWER'S CONCLUSIONS: Low molecular weight heparin is at least as effective as unfractionated heparin in preventing recurrent venous thromboembolism, and significantly reduces the occurrence of major haemorrhage during initial treatment and overall mortality at the end of follow-up. It can be adopted safely as the standard therapy for deep venous thrombosis, and studies comparing individual low molecular weight heparins are merited. PMID- 10796594 TI - Long-acting beta2-agonists for chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterised by airflow limitation which is only partially reversible. Long acting beta2 agonists, effective in the management of asthma,are also recommended for COPD management so it is important to establish whether these drugs are effective in reducing COPD symptoms in view of the potential side effect and cost burden. OBJECTIVES: To determine the effectiveness of long acting beta2-adrenoceptor agonists in improving lung function and quality of life and reducing dyspnoea in patients with COPD. SEARCH STRATEGY: A search was carried out using the Cochrane Airways Group register. Bibliographies of identified RCTs were searched for additional relevant RCTs. Authors of identified RCTs were contacted for other published and unpublished studies. In addition, unpublished studies were also obtained from the pharmaceutical companies that manufacture long acting beta2- adrenoceptor agonists. SELECTION CRITERIA: All randomised controlled trials over four weeks in duration comparing treatment with long-acting beta2-adrenoceptor agonists (salmeterol and formoterol) with placebo in patients with stable non reversible COPD. Outcome measures included forced expiratory volume in one second (FEV1), peak expiratory flow rate (PEFR), symptom scores, six minute walk distance, quality of life scores, Borg scores for dyspnoea and rescue bronchodilator use. DATA COLLECTION AND ANALYSIS: Data extraction and study quality assessment was performed independently by two reviewers. Where further or missing data were required, authors of studies were contacted. MAIN RESULTS: Thirty three abstracts were identified as potentially relevant. Of these, four randomised controlled trials (RCTs) were included in this review. Two were parallel group studies of 16 week duration and two were cross-over studies with four week treatment arms. All four RCTs assessed the efficacy of salmeterol in COPD. In a 16 week study of salmeterol 50 mcg and 100 mcg twice daily treatment, a significant increase in FEV1 was seen in both treatment groups. The weighted mean difference (WMD) for the increase in FEV1 for the 50 mcg group was 0.10 litre (95% CI: 0.05;0. 15) and in the 100 mcg group the WMD was 0.12 litre (95% CI: 0.06; 0. 17 ). In the two cross-over studies of four weeks treatment, salmeterol 50 mcg twice daily treatment did not show significant increases in FEV1 (WMD = 0.04 litre, 95% CI: -0.06; 0.15). Similarly, morning and night time PEFR was not significantly improved with salmeterol treatment. In a 16 week study, disease-specific quality of life, measured using the St. George's Respiratory Questionnaire (SGRQ), showed a significant improvement after 50 mcg twice daily, but not after 100 mcg twice daily. This improvement exceeded the threshold for a clinically significant change with this questionnaire. General health status, as measured by the Medical Outcomes Short Form 36, did not improve in any of the eight components with either salmeterol dose. No significant difference was demonstrated in the mean change from baseline in the six minute walk distance (WMD = 1.9 metres, 95% CI: -15.4;19.3). Breathlessness was reduced in one study in patients receiving salmeterol 50 mcg twice daily group. Significantly more patients in this group had Borg scores for breathlessness less than three (a score of three indicating moderate dyspnoea) compared to the placebo treated group (Peto Odds Ratio = 0.60, 95% CI: 0.40; 0.88). Neither dose of salmeterol influenced the incidence of COPD exacerbations, (50 mcg: Peto Odds Ratio = 0.74, 95% CI: 0.47, 1.15) and (100 mcg: Peto Odds Ratio = 0.98, 95% CI: 0.64, 1.52). REVIEWER'S CONCLUSIONS: Treatment of patients with COPD with long acting beta2-agonists produces only small increases in FEV1. In one study, a dose of salmeterol 50 mcg twice daily produced a reduction in breathlessness and a clinically significant improvement in quality of life. (ABSTRACT TRUNCATED) PMID- 10796595 TI - Continuous positive airways pressure for obstructive sleep apnoea. AB - BACKGROUND: Obstructive sleep apnoea (OSA) is the periodic reduction (hypopnoea) or cessation (apnoea) of breathing due to narrowing or occlusion of the upper airway during sleep. The mainstay of nocturnal treatment is continuous positive airway pressure (CPAP) therapy at home at night to prevent the resulting apnoea, hypoxia and sleep fragmentation which are believed to cause the symptoms of OSA. A number of clinical trials have evaluated the effectiveness of nasal CPAP) as a treatment. OBJECTIVES: The objective was to review the effects of CPAP in the treatment of obstructive sleep apnoea in adults. SEARCH STRATEGY: We searched Medline 1966 to 1996, Embase 1974 to 1996, Cinahl 1982 to 1995 and the reference lists of articles. We consulted experts in the field. SELECTION CRITERIA: Randomised trials comparing nocturnal CPAP with placebo or other treatments in adults with obstructive sleep apnoea and an apnoea/hypopnoea index greater than five per hour. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for missing information. MAIN RESULTS: Seven trials involving 177 people were included. All the studies had some methodological shortcomings. Six trials were of crossover design. Compared with placebo, CPAP showed significant improvements in several quality of life and depression measures. Patients preferred CPAP to placebo (odds ratio 0.4, 95% confidence interval 0.2 to 0.8). There was no significant effect on blood pressure or measures of sleepiness and sleep quality. Compared with oral appliances, CPAP significantly improved the apnoea/hypopnoea index (weighted mean difference -7.3, 95% confidence interval -10.0 to -4.7) and minimum oxygen saturation during sleep. Patients strongly preferred the oral appliance to CPAP (odds ratio 9.5, 95% confidence interval 4.3 to 21.1). REVIEWER'S CONCLUSIONS: CPAP appears to be more effective than placebo in improving some quality of life measures for people with obstructive sleep apnoea. It appears to be more effective than oral appliances in improving respiratory disturbances. People appear to prefer oral appliances over CPAP, but prefer CPAP over placebo. PMID- 10796596 TI - Inhaled steroids for episodic viral wheeze of childhood. AB - BACKGROUND: Recurrent episodic wheeze in association with viral upper respiratory tract infection (URTI) is a specific clinical illness distinct from persistent atopic asthma. OBJECTIVES: The objective of this review was to identify whether corticosteroid treatment, given episodically or daily, is beneficial to children with viral episodic wheeze. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised controlled trials (RCTs) of corticosteroid treatment versus placebo in children under 17 years of age who suffer from 'episodic viral wheeze', which is defined by wheeze in association with coryzal symptoms with minimal or no intercurrent lower respiratory tract symptoms. DATA COLLECTION AND ANALYSIS: Trial quality was assessed independently by two reviewers. Study authors were contacted for missing information. Studies were categorised according to whether treatment was given episodically or daily (maintenance). The primary outcome was episodes requiring oral corticosteroids. Secondary outcomes addressed episode severity, frequency and duration and parental treatment preference. MAIN RESULTS: Five randomised controlled trials in children with a history of mild episodic viral wheeze were identified. Most of the children had previously required no or infrequent oral corticosteroids and had very infrequent hospital admissions. There were three studies of preschool children given episodic high dose inhaled corticosteroid (1.6 - 2.25 mg per day), two using a crossover and one a parallel design. The two studies of maintenance corticosteroid (400 micrograms per day) were parallel in design, one of pre-school children the other of children aged 7 9 years. Results from the two cross-over studies of episodic high dose inhaled corticosteroids showed a reduced requirement for oral corticosteroids (Relative risk (RR)=0.53, 95% CI: 0.27, 1.04). In these 2 double blind studies, this treatment was preferred by the children's parents over placebo (RR=0.64, 95% CI: 0.48,0.87). Maintenance low dose inhaled corticosteroids did not show any clear reduction over placebo in the proportion of episodes requiring oral corticosteroids (N=2 trials, RR=0.82, 95%CI: 0.23,2.90) or in those requiring hospital admission (N=1 trial, RR=0.21, 95% CI: 0.01,4.11). REVIEWER'S CONCLUSIONS: Episodic high dose inhaled corticosteroids provide a partially effective strategy for the treatment of mild episodic viral wheeze of childhood. There is no current evidence to favour maintenance low dose inhaled corticosteroids in the prevention and management of episodic mild viral induced wheeze. PMID- 10796597 TI - Caffeine for asthma. AB - BACKGROUND: Caffeine has a variety of pharmacological effects. It is chemically related to the drug theophylline which is used to treat asthma. Accordingly, interest has been expressed in its potential role as an asthma treatment. A number of studies have explored the effects of caffeine in asthma, this is the first review to systematically examine and summarise the evidence. OBJECTIVES: Caffeine is a weak bronchodilator and it also reduces respiratory muscle fatigue. It has been suggested that caffeine may reduce asthma symptoms. The objective of this review was to assess the effects of caffeine on lung function and identify whether there is a need to control for caffeine consumption prior to lung function testing. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and the reference lists of articles. We also contacted study authors. SELECTION CRITERIA: Randomised trials of oral caffeine compared to placebo in adults with asthma. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done independently by two reviewers. MAIN RESULTS: Six trials involving a total of 55 people were included. The studies were all of cross-over design and of high quality. In comparison with placebo, caffeine appears to improve lung function for up to two hours after consumption. Forced expiratory volume in one minute showed a small improvement up to two hours after caffeine use (standardised mean difference -0.73, 95% confidence interval -1.20 to -0.25). Mid-expiratory flow rates also showed a small improvement with caffeine and this was sustained up to four hours. REVIEWER'S CONCLUSIONS: Caffeine appears to improve airways function modestly in people with asthma for up to four hours. People may need to avoid caffeine for at least four hours prior to lung function testing. PMID- 10796598 TI - Corticosteroids for pulmonary sarcoidosis. AB - BACKGROUND: Pulmonary sarcoidosis is a common condition with an unpredictable course. Oral or inhaled steroids are widely used in its treatment, but there is no consensus about when and in whom therapy should be initiated, what dose should be given and for how long. Corticosteroids given for several months have deleterious side-effects so it is important to know whether they have any maintained benefit in pulmonary sarcoidosis. OBJECTIVES: To determine the randomised controlled trial (RCT) evidence for the benefit of corticosteroids (oral or inhaled) in the treatment of pulmonary sarcoidosis. SEARCH STRATEGY: The Cochrane Airways Group interstitial lung disease RCT register was searched using the terms: sarcoidosis and (steroid* OR corticosteroid* OR prednisolone OR prednisone OR beclomethasone OR budesonide OR fluticasone). Bibliographies of retrieved RCTs and reviews were searched for additional RCTs. Pharmaceutical companies and authors of identified RCTs were contacted for other published and unpublished studies. SELECTION CRITERIA: Two reviewers independently assessed full text articles for inclusion based upon the following criteria: the study had to be a RCT or controlled clinical trial in adults with histological evidence of pulmonary sarcoidosis. DATA COLLECTION AND ANALYSIS: Study quality was assessed and data extracted independently by two reviewers. Outcomes were analysed as continuous and dichotomous outcomes, using standard statistical techniques. MAIN RESULTS: Eight RCTs were identified, two had insufficient data for any analysis. There were 338 patients in the four usable trials of oral steroids, and 66 patients in two trials of inhaled steroids. The oral steroid dose was equivalent to prednisolone 15-40 mg/day. The inhaled steroid was budesonide 0.8 - 1.2 mg/day. Outcomes were symptoms, chest X-ray (CXR) changes, lung function and global scores (a combination of all three outcomes). Oral steroids improved the CXR over 6-24 months. One study showed no improvement in lung function, in another there was an improvement in diffusing capacity in the treated group. Global scores improved in patients with stage 2 and 3 disease but not with stage 1 disease. There were no data on side-effects. Inhaled steroids had no effect on CXR. In one study diffusing capacity improved. In another, symptoms improved at the end of six months of treatment. REVIEWER'S CONCLUSIONS: Oral steroids improved the chest X-ray and a global score of CXR, symptoms and spirometry over 6-24 months. There is little evidence of an improvement in lung function. There are no data beyond 2 years to indicate whether oral steroids have any modifying effect on long-term disease progression. Oral steroids are indicated for patients with Stage 2 and 3 disease with moderate - severe or progressive symptoms or CXR changes. The available data provide no guidance for the management of this disease after 2 years. Short term (less than six months) of inhaled steroids may improved symptoms, perhaps in patients who mainly have cough. PMID- 10796599 TI - Physical training for asthma. AB - BACKGROUND: Physical training programmes have been designed for asthmatic subjects with the aim of improving physical fitness, neuromuscular coordination and self-confidence. Habitual physical activity increases physical fitness and lowers ventilation during mild and moderate exercise thereby reducing the likelihood of provoking exercise -induced asthma. Exercise training may also reduce the perception of breathlessness through a number of mechanisms including strengthening respiratory muscles. Subjectively, many asthmatics report that they are symptomatically better when fit, but results from trials have varied and have been difficult to compare because of different designs and training protocols. OBJECTIVES: The purpose of this review was to assess evidence for the efficacy and effectiveness of physical training in asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, Sportdiscus and Science citation index. SELECTION CRITERIA: Randomised trials in asthmatic subjects undertaking physical training. Subjects had to be 8 years and older. Physical training had to be undertaken for at least 20 to 30 minutes, 2 to 3 times a week, over a minimum of four weeks. DATA COLLECTION AND ANALYSIS: Eligibility for inclusion and quality of trials were assessed independently by two reviewers. MAIN RESULTS: Eight studies could be included in this review. Physical training had no effect on resting lung function or the number of days of wheeze. Physical training improved cardiopulmonary fitness as measured by an increase in maximum oxygen uptake of 5.6 ml/kg/min (95% confidence interval 3. 9 to 7.2). There were no data concerning quality of life measurements. REVIEWER'S CONCLUSIONS: In people with asthma, physical training can improve cardiopulmonary fitness without changing lung function. It is not known whether improved fitness is translated into improved quality of life. PMID- 10796600 TI - Self-management education and regular practitioner review for adults with asthma. AB - BACKGROUND: A key component of many asthma management guidelines is the recommendation for patient education and regular medical review. A number of controlled trials have been conducted to measure the effectiveness of asthma education programmes. These programmes improve patient knowledge, but their impact on health outcomes is less well established. This review was conducted to examine the strength of evidence supporting Step 6 of the Australian Asthma Management Plan: "Educate and Review Regularly"; to test whether health outcomes are influenced by education and self-management programmes. OBJECTIVES: The objective of this review was to assess the effects of asthma self-management programmes, when coupled with regular health practitioner review, on health outcomes in adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised trials of self-management education in adults over 16 years of age with asthma. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for confirmation. MAIN RESULTS: Twenty-five trials were included. Self-management education was compared with usual care in 22 studies. Self-management education reduced hospitalisations (odds ratio 0.57, 95% confidence interval 0.38 to 0.88); emergency room visits (odds ratio 0.71, 95% confidence interval (0.57 to 0.90); unscheduled visits to the doctor (odds ratio 0.57, 95% confidence interval 0.40 to 0.82); days off work or school (odds ratio 0.55, 95% confidence interval 0.38 to 0. 79); and nocturnal asthma (odds ratio 0.53, 95% confidence interval 0.39 to 0.72). Measures of lung function were little changed. Self-management programmes that involved a written action plan showed a greater reduction in hospitalisation than those that did not (odds ratio 0.35, 95% confidence interval 0.18 to 0.68). People who managed their asthma by self-adjustment of their medications using an individualised written plan had better lung function than those whose medications were adjusted by a doctor. REVIEWER'S CONCLUSIONS: Training in asthma self management which involves self-monitoring by either peak expiratory flow or symptoms, coupled with regular medical review and a written action plan appears to improve health outcomes for adults with asthma. Training programmes which enable people to adjust their medication using a written action plan appear to be more effective than other forms of asthma self-management. PMID- 10796601 TI - Self-help interventions for smoking cessation. AB - BACKGROUND: Many smokers give up smoking on their own, but provision of materials giving advice and information may assist them and increase the number who quit successfully. OBJECTIVES: The aims of this review were to determine the effectiveness of different forms of self-help materials, compared with no treatment and with other minimal contact strategies; the effectiveness of adjuncts to self-help, such as computer generated feedback, telephone hotlines and pharmacotherapy; and the effectiveness of approaches tailored to the individual compared with non-tailored materials. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register. Date of the most recent search September 1999. SELECTION CRITERIA: We included randomised trials of smoking cessation with follow-up of at least six months, where at least one arm tested a self-help intervention. We defined self-help as structured programming for smokers trying to quit without intensive contact with a therapist. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of subjects, the nature of the self-help materials, the amount of face to face contact given to subjects and to controls, outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates when available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: We identified forty-five trials. Twenty seven compared self-help materials to no intervention or tested materials as an adjunct to advice. In nine trials in which self-help was compared to no intervention there was a pooled effect which just reached statistical significance (odds ratio 1.23, 95% confidence interval 1.02 to 1.49). There was no evidence of benefit from adding self-help materials to face to face advice, or to nicotine replacement therapy. There was evidence from eight trials using materials which were tailored for the characteristics of individual smokers that such personalised materials were more effective than standard materials (odds ratio 1.41, 95% confidence interval 1.14 to 1.75). Adding follow-up telephone calls from counsellors also appeared to increase quitting (odds ratio 1.62, 95% confidence interval 1.33 to 1.97). One trial of offering access to a hotline also showed an effect. REVIEWER'S CONCLUSIONS: Self-help materials may provide a small increase in quitting compared to no intervention. There is no evidence that they have an additional benefit over other minimal interventions such as advice from a health care professional, or nicotine replacement therapy. There is evidence that materials tailored for individual smokers are more effective. PMID- 10796602 TI - Reality orientation for dementia. AB - BACKGROUND: Reality Orientation (RO) was first described as a technique to improve the quality of life of confused elderly people, although its origins lie in an attempt to rehabilitate severely disturbed war veterans, not in geriatric work. It operates through the presentation of orientation information (eg time, place and person-related) which is thought to provide the person with a greater understanding of their surroundings, possibly resulting in an improved sense of control and self-esteem. There has been criticism of RO in clinical practice, with some fear that it has been applied in a mechanical fashion and has been insensitive to the needs of the individual. There is also a suggestion that constant relearning of material can actually contribute to mood and self-esteem problems. There is often little consistent application of psychological therapies in dementia services, so a systematic review of the available evidence is important in order to identify the effectiveness of the different therapies. Subsequently, guidelines for their use can be made on a sound evidence base. OBJECTIVES: To assess the evidence of effectiveness for the use of Reality Orientation (RO) as a classroom-based therapy on elderly persons with dementia. SEARCH STRATEGY: Computerised databases were searched independently by 2 reviewers entering the terms 'Reality Orientation, dementia, control, trial or study'. Relevant websites were searched and some handsearching was conducted by the reviewer. Specialists in the field were approached for undocumented material, and all publications found were searched for additional references. SELECTION CRITERIA: All randomised controlled trials (RCTs), and all controlled trials with some degree of concealment, blinding or control for bias (second order evidence) of Reality Orientation as an intervention for dementia were included. The criteria for inclusion/exclusion involved systematic assessment of the quality of study design and the risk of bias, using a standard data extraction form. A measure of cognitive and/or behavioural change was needed. DATA COLLECTION AND ANALYSIS: Data were extracted independently by both reviewers, using a previously tested data extraction form. Authors were contacted for data not provided in the papers. Psychological scales measuring cognitive and behavioural changes were examined. MAIN RESULTS: 6 RCTs were entered in the analysis, with a total of 125 subjects (67 in experimental groups, 58 in control groups). Results were divided into 2 subsections: cognition and behaviour. Change in cognitive and behavioural outcomes showed a significant effect in favour of treatment. REVIEWER'S CONCLUSIONS: There is some evidence that RO has benefits on both cognition and behaviour for dementia sufferers. Further research could examine which features of RO are particularly effective. It is unclear how far the benefits of RO extend after the end of treatment, but and it appears that a continued programme may be needed to sustain potential benefits. PMID- 10796603 TI - Reminiscence therapy for dementia. AB - BACKGROUND: Reminiscence Therapy (RT) has been defined as vocal or silent recall of events in a person's life, either alone, or with another person or group of people. It typically involves group meetings, at least once a week. in which participants are encouraged to talk about past events, often assisted by aids such as photos, music, objects and videos of the past. There is, often, little consistent application of psychological therapies in dementia services. A number of these 'therapies' were greeted with enthusiasm by health care practitioners in understimulating care environments. They were expected to work miracles and their 'failure' to do this has led to their widespread disuse. A systematic review of the available evidence is important in order to identify the effectiveness of the different therapies. Subsequently, guidelines for their use can be made on a sound evidence base. OBJECTIVES: RT involves groups of elderly people talking of past events, assisted by aids such as videos, pictures and archives, as a means of communicating and reflecting upon their life experiences. The objective of the review is to assess the effects of RT for dementia. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, PSYCHLIT, EMBASE, OMNI, BIDS, Dissertation Abstracts International, SIGLE and reference lists of relevant articles up to 1998, and we contacted specialists in the field. We also searched relevant internet sites and we handsearched Aging and Mental Health, the Gerontologist, Journal of Gerontology, Current Opinion in Psychiatry, Current Research in Britain: Social Sciences, British Psychological Society conference proceedings and Reminiscence database. SELECTION CRITERIA: Randomised controlled trials and quasi-randomised trials of RT for dementia in elderly people. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Two trials are included in the review, but only one trial with 15 participants had extractable data. The results were statistically non-significant for both cognition and behaviour. REVIEWER'S CONCLUSIONS: No firm conclusions could be reached regarding the effectiveness of RT for dementia. The review highlighted the urgent need for more systematic research in the area. PMID- 10796604 TI - Music therapy for dementia symptoms. AB - BACKGROUND: While music/music therapy does not represent a treatment of dementia, its use is based on a possible beneficial effect on symptoms including social, emotional and cognitive skills and for decreasing behavioral problems of individuals with dementias. Thus, there are clear implications for patients' and caregivers' quality of life. However, quantification and documentation of the evidence of this effect is necessary. Professional music therapists are accountable for providing efficient, beneficial treatment. Further, music therapists are responsible for assessing, designing and implementing music therapy treatments, monitoring client progress, and reformulating their practice according to data collected and new advancements in the field. If they wait until sufficient valid, empirical data on all aspects of a disability or music response are available before attempting to design a therapy session, they may well reach retirement age before even one client can be served. On the other hand, promulgating the efficacy of music therapy in general, or of specific music therapy techniques, in the absence of any substantiation other than intuition or tradition borders on professional recklessness. OBJECTIVES: To gather and evaluate the evidence for the effectiveness of music therapy for dementia symptoms. SEARCH STRATEGY: All available sources of references were searched for randomised controlled trials of music therapy used as an intervention in dementia. The search terms included 'controlled trial or study, music*, therapy, dement*, Alzheimer*, cognitive impairment.' SELECTION CRITERIA: The reviewers assessed the methodological quality of the studies available for inclusion. The criteria used are presence and adequacy of a control condition, independent assessment of patients' performance (ie standardized ratings carried out by a person other than the music therapist) and the number of participants (no fewer than three). DATA COLLECTION AND ANALYSIS: No randomised controlled trials, or trials with quantitative data suitable for analysis were found. MAIN RESULTS: The research into music therapy to date has lacked methodological design rigour. However, the research evidence available provides sufficient grounds on which to justify further investigations into the use of music therapy in dementia patients. In this context, the reviewers discuss some of the issues and research from the studies that were considered for inclusion. REVIEWER'S CONCLUSIONS: This review was not able to identify reliable empirical evidence on which to justify the use of music therapy as a treatment for dementia. However, the evidence available suggests that music therapy may be beneficial in treating or managing dementia symptoms, and the predominant conclusion of this review is the highlighting of the need for better designed studies of the intervention. PMID- 10796605 TI - Sulpiride for schizophrenia. AB - BACKGROUND: The antipsychotic drug sulpiride was formulated over 20 years ago and was marked as having a low incidence of adverse effects and an effect on the negative symptoms of schizophrenia. This relatively inexpensive antipsychotic drug has a similar neuropharmacological profile to several novel atypical drugs. OBJECTIVES: To estimate the clinical efficacy and tolerability of sulpiride. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1997), CINAHL (1982-1998), Cochrane Schizophrenia Group's Register (March 1998), Cochrane Library (Issue 1, 1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1974 1997), SIGLE (1994-1998), and Sociofile (1974-1997) were supplemented by reference searching, contacting authors and the manufacturers of sulpiride. SELECTION CRITERIA: All randomised or quasi-randomised clinical trials focusing on the use of different doses of sulpiride or comparing sulpiride to (i) placebo; (ii) typical antipsychotic drugs; or (iii) atypical antipsychotic drugs, for those with schizophrenia or serious mental illness were selected. DATA COLLECTION AND ANALYSIS: Trials were reliably selected and quality rated. Data were independently extracted, by two reviewers (BGOS, MF), and analysed on an intention-to-treat basis. It was assumed that people who did not complete the follow up had no improvement. Authors of trials were contacted for additional and missing data. Relative risk (RR) and 95% confidence intervals (CI) of dichotomous data were calculated with the random effects model and weighted mean difference (WMD) was calculated for continuous data. MAIN RESULTS: The review currently includes 18 studies (30 citations). Studies are generally small and of poor quality. Limited evidence suggests that there is little difference between sulpiride and other drugs although the incidence of side effects may be less for sulpiride. There are no clear findings relating to negative symptoms. REVIEWER'S CONCLUSIONS: Sulpiride may be an effective antipsychotic drug but evidence is limited and data relating to claims for its value against negative symptoms is not trial-based. PMID- 10796606 TI - Seclusion and restraint for people with serious mental illnesses. AB - BACKGROUND: Seclusion and restraint are interventions used in the treatment and management of disruptive and violent behaviours in psychiatry. The use of seclusion varies widely across institutions. The literature does offer numerous suggestions for interventions to reduce or prevent aggression. OBJECTIVES: 1. To estimate the effects of seclusion and restraint compared to the alternatives for those with serious mental illnesses. 2. To estimate the effects of strategies to prevent seclusion and restraint in those with serious mental illnesses. SEARCH STRATEGY: Electronic searches of The Cochrane Controlled Trials Register (Issue 1, 1999) and The Cochrane Schizophrenia Group's Register (January 1999) were supplemented with additional searches of Biological Abstracts (1989-1999), CINAHL (1982-1999), EMbase (1980-1999), MEDLINE (1966-1999), MEDIC (1979-1999), PsycLIT (1974-1999), Sociofile (1974-1999), SPRI & SWEMED (1982-1999), Social Sciences Citation Index (1996-1999), and WILP (1983-1999). In addition, trials were sought by hand searching the reference lists of all identified studies and conference abstracts and contacting the first author of each relevant study. SELECTION CRITERIA: Randomised controlled trials were included if they focused on the use (i) of restraint or seclusion; or (ii) of strategies designed to reduce the need for restraint or seclusion in the treatment of serious mental illness. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data relative risks (RR) with 95% confidence intervals (CI) were estimated. Normal continuous data were summated using the weighted mean difference (WMD). MAIN RESULTS: 1. Effect of seclusion and restraint The search strategy yielded 2155 citations. Of these, the full articles for 35 studies were obtained. No studies met minimum inclusion criteria and no data were synthesised. Most of the 24 excluded studies focused upon the restraint of elderly, confused people and preventing them from wandering or falling. 2. Prevention of seclusion and restraint Work ongoing. REVIEWER'S CONCLUSIONS: No controlled studies exist that evaluate the value of seclusion or restraint in those with serious mental illness. There are reports of serious adverse effects for these techniques in qualitative reviews. Alternative ways of dealing with unwanted or harmful behaviours need to be developed. Continuing use of seclusion or restraint must therefore be questioned from within well-designed and reported randomised trials that are generalisable to routine practice. PMID- 10796607 TI - Zuclopenthixol decanoate for schizophrenia and other serious mental illnesses. AB - BACKGROUND: There is a clear link between stopping antipsychotic medications and a relapse of psychotic symptoms. A series of long-acting intra-muscular preparations has been developed since the 1960s in the hope of reducing the frequency of relapse and, hence, overall disability. These depot preparations, active for weeks at a time, are frequently used for those who find taking oral medication on a regular basis difficult or unacceptable. It has, however, been a consistent concern that any reduction in relapse rate afforded by depot preparations may be offset by an increase in adverse effects such as drug-induced movement disorders. OBJECTIVES: To compare zuclopenthixol decanoate to oral zuclopenthixol and other antipsychotic preparations for the treatment of schizophrenia and similar serious mental illness. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1998), CINAHL (1982-1998), The Cochrane Library (Issue 2, 1998), The Cochrane Schizophrenia Group's Register (April 1998), EMBASE (1980-1998), MEDLINE (1966-1998), and PsycLIT (1974-1998) were searched. References of all eligible studies were searched for further trials. The manufacturer of zuclopenthixol was contacted. SELECTION CRITERIA: Inclusion criteria were that the clinical study should be randomised, focus on people with schizophrenia or other serious mental illness with psychotic symptoms, and compare the use of zuclopenthixol decanoate to oral zuclopenthixol or other antipsychotic preparations. DATA COLLECTION AND ANALYSIS: Data was extracted independently by two reviewers (EC, MF). Authors of trials were contacted for additional and missing data. Odds ratios (ORs) and 95% confidence intervals (CIs) of homogenous dichotomous data were calculated with the Peto method. Where possible the number needed to treat (NNT) and its 95% confidence interval was also calculated. MAIN RESULTS: Four studies relating to zuclopenthixol decanoate were included. All compared zuclopenthixol decanoate with other depot preparations. Zuclopenthixol decanoate prevented or postponed relapses when compared to other depots (NNT 8, CI 5-53). However, zuclopenthixol decanoate may induce more adverse effects (NNH 5, CI 3-31) although it decreases need for anticholinergic medication when compared to a group of other depot preparations (NNT 9, CI 5-38). For the risk of leaving the study early, there was also a trend for benefit to those allocated to zuclopenthixol decanoate. None of the studies reported outcomes on service utilisation, costs, or quality of life. REVIEWER'S CONCLUSIONS: Choice of which depot to use must always take into account clinical judgement and the preferences of the recipients of care and their carers. Limited trial data suggests, however, that there are real differences between zuclopenthixol decanoate and other depots and these differences largely favour the former. This review highlights the need for good controlled clinical trials to fully address the effects of zuclopenthixol decanoate for those with schizophrenia. Future studies should report service utilisation data, as well as satisfaction with care and economic outcomes. Duration of such trials should be of a longer duration than the included studies (12 months or more). PMID- 10796608 TI - Interventions for treating headlice. AB - BACKGROUND: Infection with head lice is a widespread condition in developed and developing countries. Infection occurs most commonly in children, but also affects adults. If left untreated the condition can become intensely irritating and skin infections may occur if the bites are scratched. OBJECTIVES: The aim of this review was to assess the effects of interventions for head lice. SEARCH STRATEGY: Trials register of The Cochrane Infectious Diseases Group; Medline; Embase; Science Citation Index; Biosis and Toxline; reference lists of relevant articles; pharmaceutical companies producing pediculicides (published and unpublished trials); UK and US Regulatory Authorities. SELECTION CRITERIA: Randomised trials (published and unpublished) or trials using alternate allocation were sought which compared pediculicides with the same and different formulations of other pediculicides, and pediculicides with physical methods. DATA COLLECTION AND ANALYSIS: Of the 70 identified studies, only three met the inclusion criteria and one is awaiting assessment. Two reviewers independently assessed trial quality. One reviewer extracted the data. MAIN RESULTS: We found no evidence that any one pediculicide has greater effect than another. The two studies comparing malathion and permethrin with their respective vehicles showed a higher cure rate for the active ingredient than the vehicle. The third study comparing synergised pyrethrins with permethrin showed their effects to be equivalent. Adverse effects were reported in a number of trials and were all minor, although reporting quality varied between trials. REVIEWER'S CONCLUSIONS: Permethrin, synergised pyrethrin and malathion were effective in the treatment of head lice. However, the emergence of drug resistance since these trials were conducted means there is no direct contemporary evidence of the comparative effectiveness of these products. The 'best' choice will now depend on local resistance patterns. There is currently no evidence from trials to indicate the effectiveness of physical methods, such as combing/'BugBusting' or other chemical control methods such as the use of herbal treatments, when used in the curative treatment of head lice. Future trials should take into account the methodological recommendations that arise from this review. PMID- 10796609 TI - Mycobacterium vaccae immunotherapy for treating tuberculosis. AB - BACKGROUND: Mycobacterium vaccae has been advocated for immunotherapy in the treatment of tuberculosis and other infections caused by mycobacteria. OBJECTIVES: The objective of this review was to assess the effects of Mycobacterium vaccae for treating tuberculosis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials register, Medline, Embase and reference lists of articles. We also contacted organisations and individuals working in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of preparations of whole, killed Mycobacterium vaccae in patients with tuberculosis. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Four trials met the inclusion criteria. There was no effect on mortality (2 trials, RR 1.01, 95% CI 0.50 to 2.04); no consistent effect on sputum negativity or sputum culture; and a high level of adverse reactions. REVIEWER'S CONCLUSIONS: Immunotherapy with Mycobacterium vaccae does not appear to benefit patients with tuberculosis. PMID- 10796610 TI - Antibiotics for treating salmonella gut infections. AB - BACKGROUND: Antibiotic treatment of salmonella infections aims to shorten illness and prevent serious complications. There are also concerns about increasing antibiotic drug resistance. OBJECTIVES: The objective of this review was to assess the effects of antibiotics in adults and children with diarrhoea who have salmonella. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Science Citation Index, African Index Medicus, Lilacs, Extra Med and reference lists of relevant articles. We also contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing antibiotic therapy with placebo or no antibiotic therapy for salmonella infections in symptomatic or asymptomatic adults or children. Typhoid and paratyphoid salmonella infections were excluded. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data were extracted independently by two reviewers. MAIN RESULTS: Twelve trials involving 778 participants (with at least 258 infants and children) were included. There were no significant differences in length of illness, diarrhoea or fever between any antibiotic regimen and placebo. The weighted mean difference for length of illness was -0.07 days, 95% confidence interval -0.55 to 0.40; diarrhoea -0.03 days, 95% confidence interval -0.53 to 0.48; fever -0.45 days, 95% confidence interval -0. 98 to 0.08. Antibiotic regimens resulted in more negative cultures during the first week of treatment. Relapses were more frequent in those receiving antibiotics, and there were more cases with positive cultures in the antibiotic groups after three weeks. Adverse drug reactions were more common in the antibiotic groups (odds ratio 1.67, 95% confidence interval 1.05 to 2.67). REVIEWER'S CONCLUSIONS: There appears to be no evidence of a clinical benefit of antibiotic therapy in otherwise healthy children and adults with non-severe salmonella diarrhoea. Antibiotics appear to increase adverse effects and they also tend to prolong salmonella detection in stools. PMID- 10796611 TI - Interventions for treating oral lichen planus. AB - BACKGROUND: Oral lichen planus is a chronic autoimmune disease of unknown aetiology that affects the inner surface of the mouth. The symptomatic forms are painful,tend to worsen with age and with remissions being rare. Current treatment is palliative and not curative, many topical and systemic agents have been tried with little hard evidence for efficacy. OBJECTIVES: To assess the effectiveness and safety of any form of palliative therapy against placebo for the treatment of symptomatic oral lichen planus. SEARCH STRATEGY: Electronic databases, handsearching of conference proceedings and specific journals, researchers in the field, drug manufacturers. SELECTION CRITERIA: Any placebo-controlled trial of palliative therapy for symptomatic oral lichen planus, using a randomised or quasi-randomised design that measured changes in symptoms and/or clinical signs. DATA COLLECTION AND ANALYSIS: Change in symptoms (pain, discomfort) and clinical signs (visual impression, lesion measurements) at the end of therapy. Odds ratio of improvement vs no improvement for each trial outcome and pooling where appropriate. MAIN RESULTS: A total of nine RCTs were identified. The nine interventions were grouped into four separate classes (cyclosporines, retinoids, steroids and phototherapy) for comparison. No therapy was replicated exactly, the closest replication involved two trials using high and low dose cyclosporine mouthwash. Only trials recording the same outcomes in each therapeutic class were pooled. The largest number of pooled trials was three. Large odds ratios with very wide confidence intervals indicating a statistically significant treatment benefit were seen in all trials. However this has to be tempered by considerations of the small study sizes, the lack of replication, the difficulty in measuring outcome changes and the very high likelihood of publication bias. Only systemic agents were associated with treatment toxicities, all other side effects were mild and mainly limited to local mucosal reactions. REVIEWER'S CONCLUSIONS: The review provides only weak evidence for the superiority of the assessed interventions over placebo for palliation of symptomatic OLP. The results highlight the need for larger placebo-controlled RCTs with more carefully selected and standardised outcome measures before between-treatment comparisons can be properly interpreted. PMID- 10796612 TI - Amantadine and rimantadine for preventing and treating influenza A in adults. AB - OBJECTIVES: Amantadine hydrochloride and rimantadine hydrochloride have antiviral properties, but they are not widely used due to a lack of knowledge of their properties and concerns about possible adverse effects. The objective of this review was to assess the effects and safety of amantadine and rimantadine in healthy adults. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles. We also contacted manufacturers, researchers and authors. SELECTION CRITERIA: Randomised and quasi randomised studies comparing amantadine and/or rimantadine with placebo, control antivirals or no intervention, or comparing doses or schedules of amantadine and/or rimantadine in healthy adults. DATA COLLECTION AND ANALYSIS: For prevention trials the numbers of participants with clinically defined influenza, with serologically confirmed clinical influenza A and adverse effects were analysed. Analysis for treatment trials was of the mean duration of fever and adverse effects. MAIN RESULTS: Amantadine prevented 23% of clinical influenza cases (95% confidence interval 11% to 34%), and 63% of serologically confirmed clinical influenza A cases (95% confidence interval 42% to 76%) Amantadine reduced duration of fever by one day (95% confidence interval 0.7 to 1.3). Rimantadine demonstrated comparable effectiveness, but there were fewer trials and the results for prevention were not statistically significant. Both amantadine and rimantadine induced significant gastrointestinal adverse effects. Adverse effects of the central nervous system adverse and study withdrawals were significantly more common with amantadine than rimantadine. REVIEWER'S CONCLUSIONS: Amantadine and rimantadine have comparable effectiveness in the prevention and treatment of influenza A in healthy adults, although rimantadine induces fewer adverse effects than amantadine. PMID- 10796613 TI - Pharmacotherapy for inducing and maintaining remission in pouchitis. AB - OBJECTIVES: To determine the effectiveness of medical therapy (including metronidazole, bismuth carbomer enemas, oral probiotic bacteria, butyrate suppositories, and glutamine suppositories) for inducing a response or maintaining remission in pouchitis. SEARCH STRATEGY: Studies were selected using the MEDLINE data base (1966 - December 1997), abstracts from major gastrointestinal meetings and references from published articles and reviews. The Cochrane Controlled Trials Register and the Inflammatory Bowel Disease Review Group Trials Register were also searched. SELECTION CRITERIA: Four randomized controlled trials of medical therapy in adult patients with pouchitis were identified: two placebo controlled trials in active chronic pouchitis; one maintenance of remission trial comparing two active agents in chronic pouchitis; and one placebo-controlled maintenance of remission trial for chronic pouchitis. A single patient "n-of-1" trial for active chronic pouchitis was excluded. DATA COLLECTION AND ANALYSIS: Data were extracted by three independent observers based on the intention to treat principle. Each study was given a quality score based on predetermined criteria. Extracted data were converted to 2X2 tables (response versus no response and medical therapy versus placebo or medical therapy versus medical therapy) and then synthesized in to a summary statistic using the pooled odds ratio and 95% confidence intervals as described by Cochran and Mantel and Haenszel (the "odds ratio" in MetaView). MAIN RESULTS: The odds ratios of inducing a response using oral metronidazole or bismuth carbomer foam enemas compared with placebo in active chronic pouchitis were 26.67 (95% CI 2.31-308.01) and 1.00 (95% CI 0.29-3.48), respectively. The numbers needed to treat with these therapies to prevent an additional relapse was 2 for oral metronidazole, and undefined for bismuth carbomer foam enemas (due to equal efficacy between the enema and placebo). The odds ratio of maintaining remission in chronic pouchitis for oral probiotic bacteria (VSL-3) compared with placebo was 205.00 (95% CI 9.89 4247.71), while the number needed to treat to prevent one additional relapse was 2. After discontinuation of suppressive medical therapy for chronic pouchitis, there was no difference in the odds ratio of maintaining symptomatic remission with glutamine suppositories compared to butyrate suppositories, 3. 00 (95% CI 0.46-19.59). The numbers needed to treat with glutamine suppositories to prevent an additional relapse was 4. REVIEWER'S CONCLUSIONS: The results presented in this review must be interpreted with extreme caution given the small numbers of trials and patients evaluated for any one comparison. Metronidazole appears to be an effective therapy for active chronic pouchitis. Bismuth carbomer foam enemas may not be an effective therapy for chronic active pouchitis. Oral probiotic therapy with VSL-3 appears to be an effective therapy for maintaining remission in patients with chronic pouchitis in remission. There is no evidence of a difference in the maintenance of symptomatic remission in patients with chronic pouchitis treated with glutamine versus butyrate suppositories, and it is unknown whether glutamine and butyrate are equally effective or ineffective. Additional randomized, double-blind, placebo-controlled, dose-ranging clinical trials are needed to determine the efficacy of empiric medical therapies currently being used in patients with pouchitis. PMID- 10796614 TI - Topical agents or dressings for pain in venous leg ulcers. AB - BACKGROUND: Venous leg ulcers affect up to 1 per cent of people at some time in their life. These ulcers are often painful and some clinicians choose dressings to reduce the pain both during and between dressing changes. OBJECTIVES: To assess the effectiveness of dressings, local anaesthetics/analgesics for pain relief in venous leg ulceration. SEARCH STRATEGY: Cochrane Wounds Group Register and the Cochrane Collaboration Field in Complementary Medicine were searched. Cochrane Pain Palliative and Supportive Care Group and Cochrane Wounds Group strategy were combined and used. SELECTION CRITERIA: All randomised controlled trials which evaluated local interventions used to relieve venous leg ulcer pain were considered. Pain was defined as either persistent pain or pain at dressing changes or debridement. DATA COLLECTION AND ANALYSIS: Eligibility for inclusion was confirmed by two reviewers who independently assessed the all potential trials. Details of eligible studies were summarised using a data extraction sheet which was checked by the second reviewer. MAIN RESULTS: No trial evaluated interventions for persistent pain. Three trials compared a eutectic mixture of local anaesthetic (EMLA) versus placebo for pain at debridement. All 3 trials reported a significant difference in pain in favour of EMLA, however, one of the trials favoured placebo in terms of number of ulcers healed at the end of the trial and another trial noted an increased incidence of burning and itching with the use of EMLA. REVIEWER'S CONCLUSIONS: EMLA may provide pain relief for venous leg ulcer debridement however, the effect of the product on ulcer healing and the incidence of itching and burning is unclear. Research is required to address questions such as the benefits of leg ulcer debridement and the impact of EMLA on healing and the incidence of burning and itching. There were no trials addressing the treatment of persistent pain and further research is warranted. PMID- 10796615 TI - Laser therapy for venous leg ulcers. AB - OBJECTIVES: To assess the effectiveness of low level laser therapy in the treatment of venous leg ulcers. SEARCH STRATEGY: Searches of 19 databases, hand searching of journals and conference proceedings from 1948 onwards, and examination of bibliographies. SELECTION CRITERIA: Randomised controlled trials comparing low level laser therapy with: sham laser; no laser; non-coherent light. There was no restriction on date or language. The main outcome measure used was complete healing of the ulcers. DATA COLLECTION AND ANALYSIS: Data extraction was done by one reviewer and checked by a second. Meta analysis was used to combine the results of trials where the interventions and outcome measures were sufficiently similar. MAIN RESULTS: There were 4 eligible trials. Two RCTs compared laser therapy with sham, 1 with ultraviolet therapy and 1 with non coherent, unpolarised red light. Neither of the two RCTs comparing laser with sham found a significant difference in healing rates; there was no significant of laser evident when the trials were pooled. A three-arm study compared: - laser therapy alone - laser therapy plus infrared light - non-coherent, unpolarised red light. Significantly more ulcers completely healed in the group receiving a combination of laser and infrared light compared with non-coherent, unpolarised red light. A fourth trial compared laser and ultraviolet light and found no difference. REVIEWER'S CONCLUSIONS: We have found no evidence of any benefit associated with low level laser therapy on venous leg ulcer healing. One small study suggests that a combination of laser and infrared light may promote the healing of venous ulcers, however more research is needed. PMID- 10796616 TI - Nedocromil sodium for preventing exercise-induced bronchoconstriction. AB - BACKGROUND: Exercise-induced asthma causes cough, dyspnea, wheeze and chest tightness. Management of focuses on prevention through pharmaco-therapy and alternate strategies. Single use, pre-exercise beta2-agonists and non-steroidal anti-inflammatory agents such as the cromones are the most common treatments. OBJECTIVES: The objective of this review was to assess the effects of a single dose of nedocromil sodium to prevent exercise-induced bronchoconstriction. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, the Cochrane Controlled Trials Register, Current Contents, review articles, textbooks and reference lists of articles. We also contacted the drug manufacturer and primary authors for additional citations. SELECTION CRITERIA: Randomised trials comparing a single dose of nedocromil sodium with placebo to prevent exercise-induced bronchoconstriction in people over six years of age. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were conducted independently by two reviewers. Study authors were contacted for confirmation of data. MAIN RESULTS: Twenty randomised controlled trials involving 280 participants were identified. 15-60 min following inhalation of 4 mg nedocromil, the maximum fall in forced expiratory volume in one second due to exercise was improved by 15.6%, (95% CI:13.2 to 18.1) compared to the placebo response. The maximum percentage fall in peak expiratory flow rate was of the same magnitude (weighted mean difference 15.0%; 95% CI 8.3 to 21.6). Nedocromil shortened the time to recover lung normal function from more than 30 minutes with placebo to less than 10 minutes with the drug. The relative magnitude of its effect was greatest in patients with more severe exercise-induced bronchoconstriction (defined as an exercise-induced fall in lung function > 30% from baseline). There were no significant adverse effects reported. REVIEWER'S CONCLUSIONS: Nedocromil sodium used before exercise appears to reduce the severity and duration of exercise-induced bronchoconstriction. This effect appears to be more pronounced in people with severe exercise-induced bronchoconstriction. PMID- 10796617 TI - Allergen immunotherapy for asthma. AB - BACKGROUND: Allergen specific immunotherapy has long been a controversial treatment for asthma. Although beneficial effects upon clinically relevant outcomes have been demonstrated in randomised controlled trials, there remains a risk of severe and sometimes fatal anaphylaxis. The recommendations of professional bodies have ranged from cautious acceptance to outright dismissal. With increasing interest in new allergen preparations and new methods of delivery, it was time to conduct a systematic review of allergen specific immunotherapy for asthma. OBJECTIVES: Allergen specific immunotherapy involves injecting an extract of the allergen under the skin. It is also known as hyposensitisation or desensitisation, and carries a risk of potentially fatal anaphylaxis. The objective of this review was to assess the effects of allergen specific immunotherapy for asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register up to 1997, Dissertation Abstracts, WorldCat and ArticleFirst. SELECTION CRITERIA: Randomised trials using various forms of allergen specific immunotherapy to treat asthma. DATA COLLECTION AND ANALYSIS: Eligibility of studies for inclusion was decided by three reviewers independently. Quality assessment of studies was performed by two reviewers independently. MAIN RESULTS: Fifty-four trials were included. There were 25 trials of immunotherapy for house mite allergy; 13 pollen allergy trials; eight animal dander allergy trials; two Cladosporium mould allergy and six trials looking at multiple allergens. Concealment of allocation was assessed as clearly adequate in only 11 of these trials. Significant heterogeneity was present in a number of comparisons. Overall, there was a significant reduction in asthma symptoms and medication following immunotherapy. There was also a significant improvement in asthma symptom scores (standardised mean difference -0.52, 95% confidence interval -0.70 to -0.35). People receiving immunotherapy were less likely to report a worsening of asthma symptoms than those randomised to placebo (odds ratio 0.27, 95% confidence interval 0.21 to 0.35). People randomised to immunotherapy were less likely to require medication than those randomised to placebo (odds ratio 0.28, 95% confidence interval 0.19 to 0.42). Allergen immunotherapy reduced allergen specific bronchial hyper-reactivity, with some reduction in non-specific bronchial hyper-reactivity as well. There was no consistent effect on lung function. REVIEWER'S CONCLUSIONS: Immunotherapy may reduce asthma symptoms and use of asthma medications, but the size of the benefit compared to other therapies is not known. The possibility of adverse effects (such as anaphylaxis) must be considered. PMID- 10796618 TI - House dust mite control measures for asthma. AB - BACKGROUND: The major allergen in house dust comes from mites. Chemical, physical and combined methods of reducing mite allergen levels are intended to reduce asthma symptoms in people who are sensitive to house dust mites. OBJECTIVES: The objective of this review was to assess the effects of reducing exposure to house dust mite antigens in the homes of mite-sensitive asthmatics. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, checked reference lists of articles and hand-searched Respiration (1980 to 1996) and Clinical and Experimental Allergy (1980 to 1996). SELECTION CRITERIA: Randomised trials of mite control measures in asthmatic people known to be sensitive to house dust mites. DATA COLLECTION AND ANALYSIS: Two reviewers applied the trial inclusion criteria and extracted the data independently. One reviewer applied the trial quality assessment criteria. Study authors were contacted to clarify information. MAIN RESULTS: Twenty-three trials were included, with four trials awaiting assessment. There was little difference in improvement of asthma between people in experimental groups compared to control groups (odds ratio 1.2, 95% confidence interval 0.66 to 2.18). Asthma symptom scores were also similar for the experimental and control groups (standardised mean difference -0. 06, 95% confidence interval -0.54 to 0.41). These scores showed a high degree of heterogeneity. No significant difference was noted for medication usage (standardised mean difference -0.14, 95% confidence interval -0.43 to 0.15). Peak flow in the morning showed no significant difference between the experimental and the control groups (standardised mean difference -0.03, 95% confidence interval 0.25 to 0.19). REVIEWER'S CONCLUSIONS: Current chemical and physical methods aimed at reducing exposure to house dust mite allergens seem to be ineffective and cannot be recommended as prophylaxis for mite sensitive asthmatics. PMID- 10796619 TI - Nursing interventions for smoking cessation. AB - BACKGROUND: Health care professionals frequently advise patients to improve their health by stopping smoking. Such advice may be brief, or part of more intensive interventions. OBJECTIVES: To determine the effectiveness of nursing delivered smoking cessation interventions. SEARCH STRATEGY: The Cochrane Tobacco Addiction Group register was searched for studies of interventions using nurses or health visitors and an additional search made on CINAHL. SELECTION CRITERIA: Randomised trials with follow-up of at least 6 months. DATA COLLECTION AND ANALYSIS: Two authors extracted data independently. MAIN RESULTS: Fifteen studies comparing nursing intervention to a control or usual care found intervention to significantly increase the odds of quitting (Peto Odds Ratio 1.43, 95% CI 1.24 1.66). There was heterogeneity between the study results, but pooling using a random effects model did not alter the estimate of effect. There was no evidence from indirect comparison that interventions classified as intensive had a larger effect than less intensive ones. There was limited evidence that interventions were more effective for hospital inpatients with cardiovascular disease than for inpatients with other conditions. Interventions in non hospitalised patients also showed evidence of efficacy. Three studies of nurse counseling on smoking cessation during a screening health check, not included in the main meta analysis, suggested that under these conditions nursing intervention was likely to have less effect. REVIEWER'S CONCLUSIONS: The results indicate the potential benefits of smoking cessation advice and counseling given by nurses to their patients, with reasonable evidence that interventions can be effective. The challenge will be to incorporate smoking cessation intervention as part of standard practice so that all patients are given an opportunity to be queried about their tobacco use and to be given advice to quit along with reinforcement and follow-up. PMID- 10796620 TI - Donepezil for mild and moderate Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease is the most common cause of dementia and is a primary degenerative disease of the brain of unknown cause. Onset is usually late in life with increasing impairment of memory, developing gradually into a global impairment of cognition, orientation, linguistic ability and judgement. The clinical course is accompanied by growing disability and dependency on care. One of the characteristic features of the disease is the widely variable rate of progression seen in different patients. Acetylcholine is an important neurotransmitter associated with memory, and abnormalities in cholinergic neurones (including cell loss) are among the many neurological and neurochemical abnormalities that develop in AD. One approach to lessening the impact of these abnormalities is to inhibit the breakdown of acetylcholine by blocking the relevant enzyme. Tacrine was the first compound approved as a treatment for AD in the US and worked in this way, but caused severe side effects. E2020 (donepezil, Aricept) is a second generation cholinesterase inhibitor and appears to be highly specific, with relatively few side effects. OBJECTIVES: The objective of this review is to assess whether or not donepezil improves the well-being of patients with mild or moderate Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials, was searched using the terms 'donepezil', 'E2020' and 'ARICEPT'. Medline, PsychLIT and EMBASE electronic databases were searched with the above terms. Members of the Donepezil Study Group and Eisai Inc were contacted. SELECTION CRITERIA: All unconfounded, double-blind, randomised controlled trials in which treatment with donepezil was administered for more than a day and compared with placebo in patients with Alzheimer's disease. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers (JSB & DB), pooled where appropriate and possible, and the weighted or standardised mean differences or Peto odds ratios (95%CI) estimated. Where possible, intention-to-treat data were used. MAIN RESULTS: There are 4 included trials, covering treatment of 12 or 24 weeks duration in highly selected patients. The only information available on one trial (Gauthier 1998) is a conference abstract which reports no usable results. Available outcome data cover domains including cognitive function and global clinical state, but data on several important dimensions of outcome are not available. The results of three trials suggest a small beneficial effect of donepezil in improving cognitive function: at a 5mg/day dose, improvements measured -2.6 points (95%CI -3.5 -- 1.8) on weighted mean difference, in the midrange of the 70 point ADAS-Cog scale. The results of two trials show some improvement in global clinical state (assessed by an independent clinician) in those treated with donepezil compared to placebo. The patient's own rating of their Quality of Life showed no benefit of donepezil compared with placebo. There were significantly more withdrawals before the end of treatment from the 10mg/d (but not the 5mg/d) donepezil group compared with placebo, which may have resulted in some overestimation of beneficial changes at 10mg/d in progressively declining characteristics, as last available measures were used in analyses. A variety of adverse effects were recorded, but very few patients left a trial as a direct result of the intervention. REVIEWER'S CONCLUSIONS: In selected patients with mild or moderate Alzheimer's disease treated for periods of 12 or 24 weeks, donepezil produced modest improvements in cognitive function and study clinicians rated global clinical state more positively in treated patients. No improvements were present on patient self-assessed quality of life and data on many important outcomes are not available. The practical importance of these changes to patients and carers is unclear. PMID- 10796621 TI - Rivastigmine for Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease (AD) is the most common cause of dementia in the elderly. One of the most successful therapeutic strategies for Alzheimer's disease has been the use of acetylcholinesterase inhibitors to enhance surviving cholinergic neurotransmission by inhibiting breakdown of released acetylcholine. The first generation acetylcholinesterase inhibitors, such as tacrine, revealed major limitations to use including hepatotoxicity. Several second generation acetylcholinesterase inhibitors have now been introduced, including rivastigmine, which are believed to have superior proprieties. The mode of action and metabolism of rivastigmine suggest that it is unlikely to interact significantly with other medications. This is of particular relevance in elderly AD patients, the majority of whom are likely to be receiving concomitant medication. Large multi-centre trials have been completed in the USA, Canada, Europe and South Africa. Rivastigmine has received EU approval for use in all member states. It has approval in 30 countries but not the US. It is currently under review by the Food and Drug Administration, who requested additional analyses in 1998. OBJECTIVES: To determine the clinical efficacy and safety of rivastigmine for patients with dementia of the Alzheimer's type. SEARCH STRATEGY: The Cochrane Controlled Trials Register, the Dementia Group Register of Clinical Trials, other electronic databases and other sources of reports were searched using the terms ENA 713, EXELON, and rivastigmine in addition to the terms for controlled trials in dementia (see the Group's search strategy for full details). SELECTION CRITERIA: All unconfounded, double-blind, randomised trials in which treatment with rivastigmine was administered for more than one day and compared to placebo for patients with dementia of the Alzheimer's type. DATA COLLECTION AND ANALYSIS: Data were extracted by the reviewer (JSB) and entered into an appropriate meta analysis. The data extracted were cross-checked by the second reviewer (VI). For each outcome measure, data were sought on every patient randomised. To allow an intention-to-treat analysis, the data were sought irrespective of compliance, whether or not the patient was subsequently deemed ineligible, or otherwise excluded from treatment or follow-up. If these data were not available, an analysis of data on patients who completed treatment was conducted. MAIN RESULTS: There are seven included trials. There are no published reports for two large phase III trials, B304 and B351, although they were completed more than 3 years ago. These are part of the Novartis ADENA programme and comprise 1379 (49%) out of 2803 phase III patients. It is unclear how missing data are replaced in ITT analyses, as reports from the ADENA programme provide no description of the use of this method. This has a profound effect on the results: if the method is substantially the same as LOCF, the benefits of treatment inferred from the analyses described in the publications as ITT, may be exaggerated. The meta analysis reveals benefits on cognitive function as measured by ADAS-Cog test scores for the higher dose of rivastigmine compared to placebo at 26 weeks and for the lower dose. An additional analysis of ADAS-Cog dichotomised into those showing less than 4 points improvement and those showing 4 or more points improvement at 26 weeks shows benefit for cognitive function for the higher dose of rivastigmine compared to placebo and not for the lower dose. Global clinical state, dichotomised, counting those showing no change or decline, against those showing improvement shows benefit due to lower dose rivastigmine compared to placebo at 26 weeks and not for the higher dose. One trial reported results at 18 weeks and there are no significant differences between higher dose rivastigmine and placebo. One trial reported results at 13 weeks, and there are no significant differences between the 4 or 6 mg/d rivastigmine group and p PMID- 10796622 TI - Polyunsaturated fatty acid (fish or evening primrose oil) for schizophrenia. AB - BACKGROUND: Limited evidence gives support to an hypothesis suggesting that the symptoms of schizophrenia may result from altered neuronal membrane structure and metabolism. The latter are dependent on blood plasma levels of certain essential fatty acids (EFAs) and their metabolites. Several studies have shown those with schizophrenia often have low levels of the particular EFAs necessary for normal nerve cell membrane metabolism. OBJECTIVES: To review the effects of supplementing standard antipsychotic treatment with polyunsaturated fatty acids, whether essential (EFAs) or non-essential, for those with schizophrenia and, in recent updates to also evaluate the effects of EFA's as a sole antipsychotic treatment. To evaluate the relative efficacy of different types of fatty acid supplementation. SEARCH STRATEGY: Relevant randomised trials were identified by searching the following electronic databases: Biological Abstracts (1985-1998), CINAHL (1982-1998), Cochrane Library (Issue 4, 1999), Cochrane Schizophrenia Group's Register (February 2000), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1974-1998). In addition, reviewers searched references of included and excluded studies and contacted authors to identify further studies. SELECTION CRITERIA: All randomised clinical trials of polyunsaturated fatty acid supplementation to standard treatment or as primary intervention for schizophrenia (however defined) versus standard care. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed on an intention-to treat basis. They assumed that people who left the study early or were lost to follow-up had no improvement. Where possible and appropriate relative risk (RR) and their 95% confidence intervals (CI) were calculated. The number needed to treat (NNT) was estimated. For continuous data weighted mean differences (WMD) and their 95% confidence intervals were calculated. Data were inspected for heterogeneity and publication biases. MAIN RESULTS: Four relatively small trials (total n=204) showed low levels of loss to follow up and adverse effects for those taking essential fatty acids. Early results from a few trials suggest a positive effect of eicosapentaenoic acid (EPA) over placebo for scale-derived mental state outcomes. The data, however, is limited making these results difficult to analyse and interpret with confidence. A single small study (n=30) investigated the value of using EPA as sole treatment for people hospitalised for relapse. Results suggest that EPA may help one third of people avoid instigation of standard antipsychotic drugs for 12 weeks (RR 0.6, CI 0.4-0.91). There were no clear effects of primrose oil (omega-6) EFA supplementation. REVIEWER'S CONCLUSIONS: All data are preliminary, but results look encouraging for fish oil. EPA does not seem harmful, may be acceptable to people with schizophrenia and have moderately positive effect. A further trial is soon to be reported from the USA and more are underway or planned in the South Africa and Norway. Considering that EPA may be an acceptable intervention, large, long simple studies reporting clincially meaningful data should be anticipated. PMID- 10796623 TI - Vaccines for preventing typhoid fever. AB - BACKGROUND: Whole cell vaccines, consisting of relatively crude preparations of Salmonella typhi administered parenterally, are effective but have a high incidence of adverse effects. Two vaccines have been developed more recently. Ty21a (an attenuated strain of S. typhi administered orally) and Vi (the purified bacterial capsule, given parenterally), have appeared less toxic than the older whole cell vaccines and are thought to be equally effective. OBJECTIVES: The objective of this review was to assess the effects of typhoid fever vaccines. SEARCH STRATEGY: We searched the Cochrane Library, Medline, Index Medicus, Embase and reference lists of articles. SELECTION CRITERIA: Randomised trials comparing typhoid vaccines to other types of vaccine or placebo. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Seventeen studies, involving nearly two million people, were included. For the whole cell vaccines single dose regimens provided significant protection for the first two years. Two dose regimens provided significant protection for five years. For the Ty21a vaccine, both two and three dose regimens provided statistically significant protection for two years. The three dose regimen provided protection in the third and fourth years, but protection was not statistically significant in the fifth year. The Vi vaccine provided protection for two years, but the protection in the third year was not significant. The three year cumulative efficacy of two doses of whole cell vaccines was 73% (95% confidence interval 65-80), three doses of Ty21a was 51%, (95% confidence interval 35 to 63) and one dose of Vi was 55% (95% confidence interval 30 to 71). Data on adverse effects were limited, but indicate that whole cell vaccines are more toxic than the newer Ty21a and Vi vaccines. REVIEWER'S CONCLUSIONS: The whole cell vaccines provided more prolonged protection than either the Ty21a vaccine or the Vi vaccine. However whole cell vaccines are associated with higher toxicity. PMID- 10796624 TI - Rice-based oral rehydration solution for treating diarrhoea. AB - BACKGROUND: Oral rehydration therapy is used to treat dehydration caused by diarrhoea. However the rehydration solution does not reduce stool loss or length of illness. A solution able to do this may lessen the use of ineffective diarrhoea treatments as well as improve morbidity and mortality related to diarrhoea. OBJECTIVES: The objective of this review was to assess the effects of rice-based oral rehydration salts solution compared with glucose-based oral rehydration salts solution on reduction of stool output and duration of diarrhoea in patients with acute watery diarrhoea. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase, Lilacs and the reference lists of relevant articles. We also contacted researchers in the field. SELECTION CRITERIA: Randomized trials comparing standard World Health Organization oral rehydration solution with an experimental oral rehydration salts solution in which glucose (20 grams per litre) was replaced by 50-80 grams per litre of rice powder, with the electrolytes remaining unchanged. DATA COLLECTION AND ANALYSIS: Data were extracted independently by a statistician and a clinician. MAIN RESULTS: Twenty two trials were included. Concealment of allocation was adequate in 15 of these trials. Irrespective of age, people with cholera who were given rice oral rehydration salts solution had substantially lower rates of stool loss than those given oral rehydration salts solution in the first 24 hours. Mean stool outputs in the first 24 hours were lower by 67 millilitres/kg of body weight (weighted mean difference -67.4, 95% confidence interval -94.3 to -41.0) in children, and by 51 millilitres/kg of body weight (weighted mean difference -51.1, 95% confidence interval -65.9 to -36.3) in adults. The rate of stool loss in infants and children with acute non-cholera diarrhoea was reduced by only four millilitres/kg of body weight (weighted mean difference -4.3, 95% confidence interval -9.3 to 0.8). REVIEWER'S CONCLUSIONS: Rice-based oral rehydration appears to be effective in reducing stool output in people with cholera. This effect was not apparent in infants and children with non-cholera diarrhoea. PMID- 10796625 TI - Neuraminidase inhibitors for preventing and treating influenza in healthy adults. AB - OBJECTIVES: Influenza is an acute respiratory infection associated with raised temperature, headache, muscle ache and cough. The objective of this review was to assess the effects of neuraminidase inhibitors (NIs) in preventing cases of influenza and shortening or reducing the severity of influenza in healthy adults. A further objective was to estimate the frequency of adverse effects associated with NI administration. SEARCH STRATEGY: We searched Medline, the Cochrane Acute Respiratory Infections Group trials register, the Cochrane Controlled Trials Register (CCTR), manufacturers' databases, Embase (1991 to 1998) and reference lists of articles in May 1999. We also contacted manufacturers, researchers in the field, and authors of studies evaluated in the review. SELECTION CRITERIA: Randomised or quasi-randomised placebo-controlled studies of NIs in healthy adults. Studies assessing protection or treatment from exposure to naturally occurring and experimental influenza were considered. The main outcomes were numbers and/or severity of influenza cases and the number and seriousness of adverse effects. DATA COLLECTION AND ANALYSIS: Two reviewers applied the inclusion criteria to the retrieved studies, assessed trial quality and extracted data. MAIN RESULTS: Eight trials with 1180 adults were included. Overall the methodological quality of the studies appeared to be good. As a preventive measure, NIs when compared to placebo were 74% effective (95% confidence interval 50% to 87%) in preventing naturally occurring cases of clinically defined influenza, and 60% effective (95% confidence interval 76% to 33%) in preventing cases of laboratory confirmed influenza. As a treatment, NIs shorten the duration of symptoms by one day - weighted mean difference 1 (95% confidence interval -1.3 to -0.6). The time gained in returning to normal activities is half a day - weighted mean difference -0.5 ( 95% confidence interval -1.1 to -0.1) for laboratory cases of influenza. The adverse event profile (local nasal irritation) of Zanamivir appears no better than placebo - odds ratio 1.19 ( 95% confidence interval 0.39 to 3.62). Compared with rimantadine in a preventive role, Oseltamivir has a significantly lower incidence of adverse effects and significantly higher incidence of nausea. For treatment, the adverse event profile shows that gastrointestinal symptoms are significantly worse in NIs than placebo - Peto odds ratio 2.32 ( 95% confidence interval 1.55 to 3.47). REVIEWER'S CONCLUSIONS: NIs are effective for the prevention and treatment of influenza. Overall NIs are safe, although Oseltamivir causes significant nausea. PMID- 10796626 TI - Bronchodilators for bronchiolitis. AB - OBJECTIVES: Bronchiolitis is an acute, highly communicable lower respiratory tract infection. Bronchodilators are commonly used in the management of bronchiolitis in North America, but not in the United Kingdom. The objective of this review was to assess the effects of bronchodilators for bronchiolitis. SEARCH STRATEGY: We searched MEDLINE, EMBASE, Reference Update, reference lists of articles, and the files of two of the authors up to June 1998. SELECTION CRITERIA: Randomised trials comparing bronchodilators with placebo in the treatment of bronchiolitis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Unpublished data were requested from authors when necessary. MAIN RESULTS: In eight trials with 394 children, 46% demonstrated an improved clinical score with bronchodilators compared to 75% with placebo (odds ratio for no improvement 0.29, 95% confidence interval 0.19 to 0.45). However, the inclusion of studies that enrolled people with recurrent wheezes may have biased these results in favour of bronchodilators. Bronchodilator recipients did not show improvement in measures of oxygenation, the rate of hospitalisation (18% versus 26%, odds ratio 0.70, 95% confidence interval 0.36 to 1.35) or duration of hospitalisation (weighted mean difference 0.12, 95% confidence interval -0.3 to 0.5). REVIEWER'S CONCLUSIONS: Bronchodilators produce modest short-term improvement in clinical scores. This small benefit must be weighed against the costs of these agents. PMID- 10796627 TI - Chest radiograph in acute respiratory infections in children. AB - BACKGROUND: Chest radiography is widely used in acute lower respiratory infection in children, but the benefits are unknown. OBJECTIVES: To assess the effects of chest radiography for children with acute lower respiratory infections. SEARCH STRATEGY: We searched the Cochrane Respiratory Infections Group trials register, the Cochrane Controlled Trials Register and MEDLINE up to December 1999. We contacted experts in the fields of acute respiratory infections and paediatric radiology to locate additional studies. SELECTION CRITERIA: Randomised or quasi randomised trials of chest radiography in acute respiratory infections in children. DATA COLLECTION AND ANALYSIS: One reviewer extracted data and assessed trial quality. MAIN RESULTS: We identified only one trial of 522 participants, performed by the reviewers. The participants were ambulatory children aged two months to five years. Forty six percent of both radiography and control participants had recovered by seven days - odds ratio (1.03, 95% confidence interval 0.64 to 1.64). Thirty three percent of radiography participants and 32% of control participants made a subsequent hospital visit within four weeks - odds ratio 1.02 (95% confidence interval 0.71 to 1.48). Three percent of both radiography and control participants were subsequently admitted to hospital within four weeks - odds ratio 1.02 (95% confidence interval 0.40 to 2.60). There were no deaths in either group. REVIEWER'S CONCLUSIONS: There is no evidence that chest radiography improves outcome in ambulatory children with acute lower respiratory infection. The findings do not exclude a potential effect of radiography, but the potential benefit needs to be balanced against the hazards and expense of chest radiography. The findings apply to ambulatory children only. PMID- 10796628 TI - Vaccines for preventing influenza in healthy adults. AB - OBJECTIVES: To identify, retrieve and assess all studies evaluating the effects of vaccines on influenza in healthy adults. To assess the effectiveness of vaccines in preventing cases of influenza in healthy adults. To estimate the frequency of adverse effects associated with influenza vaccination in healthy adults. SEARCH STRATEGY: MEDLINE was searched using the strategy of the Cochrane Acute Respiratory Infections Group. The bibliography of retrieved articles, the Cochrane Controlled Trials Register (CCTR), and EMBASE (1990 to 1997) were also searched. Handsearch of the journal Vaccine from its first issue to the end of 1997 (Jefferson and Jefferson, 1996; Jefferson, 1998). We wrote to vaccine manufacturers and first or corresponding authors of studies in the review. SELECTION CRITERIA: Any randomised or quasi-randomised studies comparing influenza vaccines in humans with placebo, control vaccines or no intervention, or comparing types, doses or schedules of influenza vaccine. Live, attenuated or killed vaccines or fractions thereof administered by any route, irrespective of antigenic configuration were considered. Only studies assessing protection from exposure to naturally occurring influenza in healthy individuals aged 14 to 60 (irrespective of influenza immune status) were considered. DATA COLLECTION AND ANALYSIS: Both clinically defined cases and serologically confirmed cases of influenza were considered as outcomes according to the authors' definitions. Time off work, complication and hospitalisation rates were considered, together with adverse effects. Vaccine schedules were analysed including one component matching the recommended vaccine (WHO or government recommendations) for the year of the study, and whether they matched the circulating viral subtypes. MAIN RESULTS: The recommended live aerosol vaccines reduced the number of cases of serologically confirmed influenza A by 48% (95% confidence interval 24% to 64%), whilst recommended inactivated parenteral vaccines had a vaccine efficacy of 68% (95% confidence interval 49% to 79%). The vaccines were less effective in reducing clinical influenza cases, with efficacies of 13% and 24% respectively. Use of the vaccine significantly reduced time off work, but only by 0.4 days for each influenza episode (95% confidence interval 0.1 to 0.8 days). Analysis of vaccines matching the circulating strain gave higher estimates of efficacy, whilst inclusion of all other vaccines reduced the efficacy. REVIEWER'S CONCLUSIONS: Influenza vaccines are effective in reducing serologically confirmed cases of influenza A. However, they are not as effective in reducing cases of clinical influenza. The use of WHO recommended vaccines appears to enhance their effectiveness in practice. PMID- 10796629 TI - Oral zinc for arterial and venous leg ulcers. AB - BACKGROUND: Leg ulcers affect up to 1 per cent of people at some time in their life. Management includes care of the ulcer using dressings and treatment of underlying medical problems such as malnutrition, lack of minerals, vitamins, poor blood supply or infection. OBJECTIVES: To assess the effectiveness of oral zinc in healing arterial or venous leg ulcers. SEARCH STRATEGY: Searches of 19 databases, hand searching of journals and conference proceedings from 1948 onwards, and examination of bibliographies. The company manufacturing zinc sulphate tablets was asked for references to relevant trials. SELECTION CRITERIA: Randomised controlled trials comparing oral zinc sulphate with placebo or no treatment in patients with arterial or venous leg ulcers. There was no restriction on date or language. The main outcome measure used was complete healing of the ulcers. Trials were eligible for inclusion if they measured ulcer healing objectively, by time to complete healing, proportion of ulcers healed during the study, or healing rates of ulcers. DATA COLLECTION AND ANALYSIS: All data extraction and assessment of trial quality were done by both authors independently. MAIN RESULTS: There were six eligible trials. All are small and serum zinc was measured at baseline or during the trial in 4 trials. Overall there is no evidence of a beneficial effect of treatment with zinc sulphate on the number of ulcers healed at the end of the trials. There is some evidence that oral zinc might have a beneficial effect on healing of venous ulcers in people with a 'low' serum zinc level at baseline. REVIEWER'S CONCLUSIONS: Overall, oral zinc sulphate does not appear to aid healing of leg ulcers, although it might be beneficial in those with venous leg ulcers and low serum zinc. Further research is needed to ascertain the serum zinc concentration below which treatment with zinc might be beneficial, and the dose required. [This abstract was prepared centrally] PMID- 10796630 TI - Anti-cholinergic drugs for wheeze in children under the age of two years. AB - BACKGROUND: Wheeze in infancy and early childhood is common and appears to be increasing though the magnitude of any increase is unclear. Most wheezing episodes in infancy are precipitated by respiratory viral infections. Treatment of very young children with wheeze remains controversial. Anti-cholinergics are often prescribed but practice varies widely and the efficacy of this form of therapy remains the subject for debate. OBJECTIVES: Wheeze in infancy and early childhood is common and appears to be increasing. Most wheezing episodes in infancy are a result of viral infection. Bronchodilator medications such as beta2 agonists and anti-cholinergic agents are often used to relieve symptoms, but patterns of use vary. The objective of this review was to assess the effects of anti-cholinergic therapy in the treatment of wheezing infants. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and the reference lists of articles. We contacted researchers in the field and industry sources. SELECTION CRITERIA: Randomised trials that compared anti-cholinergic therapy with placebo or beta2-agonists in wheezing children under two years of age. Children with acute bronchiolitis and chronic lung disease were excluded. DATA COLLECTION AND ANALYSIS: Eligibility for inclusion and quality of trials were assessed independently by two reviewers. MAIN RESULTS: Six trials involving 321 infants in three different settings were included. Compared with beta2-agonist alone, the combination of ipratropium bromide and beta2-agonist was associated with a reduced need for additional treatment, but no difference was seen in treatment response, respiratory rate or oxygen saturation improvement in the emergency department. There was no significant difference in length of hospital stay between ipratropium bromide and placebo; or between ipratropium bromide and beta2 agonist combined compared with beta2-agonist alone. However, combined ipratropium bromide and beta2-agonist compared to placebo showed significantly improved clinical scores at 24 hours. Parents preferred ipratropium bromide over nebulised water or placebo for relief of their children's symptoms at home. REVIEWER'S CONCLUSIONS: There is not enough evidence to support the uncritical use of anti cholinergic therapy for wheezing infants, although parents using it at home were able to identify benefits. PMID- 10796631 TI - Long acting beta-agonists versus theophylline for maintenance treatment of asthma. AB - BACKGROUND: Theophylline and long acting beta2-agonists are bronchodilators used for the management of persistent asthma symptoms, especially nocturnal asthma. They represent different classes of drug with differing side-effect profiles. OBJECTIVES: To assess the comparative efficacy, safety and side-effects of long acting beta-agonists and theophylline in the maintenance treatment of asthma. SEARCH STRATEGY: Randomised, controlled trials (RCTs) were identified using the Cochrane Airways Group register. The register was searched using the following terms: asthma and theophylline and long acting beta-agonist or formoterol or foradile or eformoterol or salmeterol or bambuterol or bitolterol. Titles and abstracts were then screened to identify potentially relevant studies. The bibliography of each RCT was searched for additional RCTs. Authors of identified RCTs were contacted for other relevant published and unpublished studies. SELECTION CRITERIA: All included studies were RCTs involving adults and children with clinical evidence of asthma. These studies must have compared oral sustained release and/or dose adjusted theophylline with an inhaled long-acting beta agonist. DATA COLLECTION AND ANALYSIS: Potentially relevant trials, identified by screening titles and/or abstracts, were obtained. Two reviewers independently assessed full text versions of these trials to decided whether the trial should be included in the review, and assessed its methodological quality. Where there was disagreement between reviewers, this was resolved by consensus, or reference to a third party. Data were extracted by two independent reviewers. Inter-rater reliability was assessed by simple agreement. Study authors were contacted to clarify randomisation methods, provide missing data, verify the data extracted and identify unpublished studies. Relevant pharmaceutical manufacturers were also contacted. MAIN RESULTS: Six trials met the inclusion criteria. Five used salmeterol and one, biltoterol. They were of varying quality. There was a trend for salmeterol to improve FEV1 more than theophylline in three studies and salmeterol use was associated with more symptom free nights. Bitolterol, used in only one study, was reported to be less effective than theophylline. Subjects taking salmeterol experienced fewer adverse events than those using theophylline (Relative Risk 0.38; 95%Confidence Intervals 0.25, 0.57). Significant reductions were reported for central nervous system adverse events (Relative Risk 0.51; 95%Confidence Intervals 0.30, 0.88) and gastrointestinal adverse events (Relative Risk 0.32; 95%Confidence Intervals 0.17, 0.59). REVIEWER'S CONCLUSIONS: Salmeterol may be more effective than theophylline in reducing asthma symptoms including night waking and improving lung function. More adverse events occurred in subjects using theophylline when compared to salmeterol. PMID- 10796632 TI - Beclomethasone for asthma in children: effects on linear growth. AB - BACKGROUND: Inhaled steroids play a central role in the management of childhood asthma. There is concern about their side effects, especially on growth. However asthma may also cause growth retardation. Growth rates are not stable, so randomised controlled parallel group studies are needed to assess the impact of inhaled steroids on growth. This review is confine to one inhaled steroid, beclomethasone, that is known to have significant levels of systemic absorption. OBJECTIVES: To determine whether inhaled beclomethasone cause significant delay in the linear growth of children with asthma. SEARCH STRATEGY: The Cochrane Airways Group asthma register was searched. Bibliographies from included studies, and known reviews were searched for additional citations. Personal contact with colleagues and researchers working in the field of asthma were made to identify potentially relevant trials. SELECTION CRITERIA: Randomized, controlled trials comparing the effects of beclamethasone to non-steroidal medication (placebo or non-steroidal therapy) on the linear growth of children with asthma. DATA COLLECTION AND ANALYSIS: Data related to the clinical outcome "change in growth" were extracted by two reviewers working independently MAIN RESULTS: One hundred and fifty-nine citations were identified by the search strategy and bibliography review. Three studies met the inclusion criteria. All used beclomethasone 200 mcg twice daily delivered by dry powder Diskhaler to treat children with mild moderate asthma. Study duration was 7-12 months. In all three studies, a significant decrease in linear growth occurred in children treated with beclomethasone compared to those receiving placebo or non-steroidal asthma therapy. The average decrease, calculated through meta-analysis, was -1.54 cm per year (95% CI -1.15, -1.94). REVIEWER'S CONCLUSIONS: In children with mild moderate asthma, beclomethasone 200 mcg twice daily caused a decrease in linear growth of -1.54 cm per year. These studies lasted a maximum of 54 weeks, so it remains unclear whether the decrease in growth is sustained or whether it reverses with 'catch up' after therapy is discontinued. We are unable to comment on growth effects of other inhaled steroids that have potentially less systemic effects. If inhaled steroids are required to control a child's asthma, we recommend using the minimum dose that effectively controls the child's asthma and closely following growth. PMID- 10796633 TI - Dietary marine fatty acids (fish oil) for asthma. AB - BACKGROUND: Epidemiological studies suggest that a diet high in marine fatty acids (fish oil) may have beneficial effects on inflammatory conditions such as rheumatoid arthritis and possibly asthma. OBJECTIVES: 1. To determine the effect of marine n-3 fatty acid (fish oil) supplementation in asthma. 2. To determine the effect of a diet high in fish oil in asthma. SEARCH STRATEGY: The Cochrane Airways Review Group register was search using the terms: marine fatty acids OR diet OR nutrition OR fish oil OR eicosapentaenoic acid OR EPA. Bibliographies of retrieved trials were searched and fish oil manufacturers contacted. SELECTION CRITERIA: Randomised controlled trials in patients with asthma more than two years of age were included. The study duration had to be in excess of 4 weeks. Double blind trials were preferred, but single-blind and open trials were also reviewed for possible inclusion. Three reviewers read each paper, blind to its identity. Decisions concerning inclusion were made by simple majority. Quality assessment was performed by all three reviewers independently. DATA COLLECTION AND ANALYSIS: The only comparison possible was between marine n-3 fatty acid supplementation and placebo. There were insufficient trials to examine dietary manipulation alone. MAIN RESULTS: Eight randomised controlled trials conducted between 1986 and 1998 satisfied the inclusion criteria. Six were of parallel design and two were cross-over studies. Seven compared fish oil with placebo whilst one compared high dose vs low dose marine n-3 fatty acid supplementation. None of the included studies reported asthma exacerbations, health status or hospital admissions. There was no consistent effect on any of the analyzable outcomes: FEV1, peak flow rate, asthma symptoms, asthma medication use or bronchial hyper reactivity. The single study performed in children also combined dietary manipulation with fish oil supplementation and showed improved peak flow and reduced asthma medication use. There were no adverse events associated with fish oil supplements. REVIEWER'S CONCLUSIONS: There is little evidence to recommend that people with asthma supplement or modify their dietary intake of marine n-3 fatty acids (fish oil) in order to improve their asthma control. Equally, there is no evidence that they are at risk if they do so. PMID- 10796634 TI - Mucolytic agents for chronic bronchitis or chronic obstructive pulmonary disease. AB - BACKGROUND: Individuals with chronic bronchitis or chronic obstructive pulmonary disease (COPD) may suffer recurrent exacerbations with an increase in volume and/or purulence of sputum and any therapy that reduced the number of exacerbations would be useful. There is a marked difference between countries in terms of the prescribing of mucolytics depending on whether or not they are perceived to be effective. OBJECTIVES: To assess the effects of oral mucolytics in adults with stable chronic bronchitis or COPD. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised trials that compared oral mucolytic therapy with placebo for at least two months in adults with chronic bronchitis or COPD. Studies of people with asthma and cystic fibrosis were excluded. DATA COLLECTION AND ANALYSIS: One reviewer extracted data. Study authors and drug companies were contacted for missing information. MAIN RESULTS: Twenty two trials were included. Compared with placebo, there was a significant reduction in the number of exacerbations per patient with oral mucolytics (weighted mean difference (WMD) 0.067 per month, 95% confidence interval -0.079, -0.055, p<0.001) which is a 29% reduction. The annualised rate of exacerbations in the control patients was 2.7 per year. The number of days of disability also fell on mucolytic therapy (WMD 0.56, 95% confidence interval -0.77, -0.35, p<0.001). The number of patients who remained exacerbation-free was greater in the mucolytic group thanin the placebo group (OR 2.22, 95% confidence interval 1.93, 2.54, p<0.001). There was no difference in lung function or in adverse effects reported between treatments. REVIEWER'S CONCLUSIONS: In subjects with chronic bronchitis or COPD, treatment with mucolytics was associated with a small reduction in acute exacerbations and a somewhat greater reduction in total number of days of disability. PMID- 10796635 TI - Corticosteroids for acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Acute exacerbations occur quite commonly in patients with chronic obstructive pulmonary disease (COPD). Corticosteroid drugs, either parenteral or oral, are used commonly in this setting. OBJECTIVES: To determine the effect of corticosteroids, administered either parenterally or orally, on the outcome in patients with acute exacerbations of COPD. SEARCH STRATEGY: An initial search was carried out using the Cochrane Airways Group COPD register with additional studies sought in the bibliographies of randomised controlled trials and review articles. Authors of identified randomised controlled trials were contacted for other published and unpublished studies. SELECTION CRITERIA: Randomised controlled trials comparing corticosteroids, administered either parenterally or orally, with appropriate placebo. Other interventions were standardised e.g. bronchodilators, antibiotics. Studies of acute asthma were excluded. DATA COLLECTION AND ANALYSIS: Data was extracted by one reviewer and sent to authors for verification. All trials were combined for analysis where possible. MAIN RESULTS: We identified 7 studies that fulfilled the inclusion criteria. Outcomes were varied and few were common to all studies. The most commonly reported outcome, the FEV1 between 6 - 72 hours after treatment, showed no significant difference between corticosteroid and placebo treatment. Treatment failure (defined as re-attendance in the emergency department, need for oral steroids or hospitalisation) and quality of life did show a statistically significant benefit for corticosteroid treatment, but the number of studies reporting these outcomes was small and there was significant heterogeneity between them REVIEWER'S CONCLUSIONS: Treatment with oral or parenteral corticosteroids in outpatients may decrease the number of patients requiring further treatment or hospitalisation, but otherwise it has no significant effect on the outcome of acute exacerbations of chronic obstructive airways disease. Further research is required to determine the place of corticosteroid treatment in acute exacerbations of chronic obstructive airways disease. PMID- 10796636 TI - Mucolytics for bronchiectasis. AB - BACKGROUND: Bronchiectasis is usually characterised by the production of large quantities of sputum that patients frequently have difficulty in expectorating. Mucolytic agents target hyper-secretion or changed physiochemical properties of sputum to make it easier to clear. One drug, recombinant human DNase, breaks down the DNA that is released at the site of infection by neutrophils. OBJECTIVES: The objective of this review was to assess the effects of ingested or inhaled mucolytics in patients with bronchiectasis. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, reference lists of relevant articles. We also contacted experts in the field and drug companies. SELECTION CRITERIA: Randomised trials of mucolytic treatment in people with bronchiectasis but not cystic fibrosis. DATA COLLECTION AND ANALYSIS: Data extraction was performed independently by two reviewers. Study authors were contacted for confirmation. MAIN RESULTS: Two trials were included. In one study, compared to placebo, high doses of bromhexine combined with antibiotics eased difficulty in expectoration (weighted mean difference -0.53, 95% confidence interval -0.81 to -0.25 at 16 days). There was also a reduction in sputum production with bromhexine (weighted mean difference -21.5%, 95% confidence interval -38.9 to -4.1 at day 16). There was no difference in forced expiratory volume. In a second study, compared to placebo, recombinant human DNase showed no difference in forced expiratory volume or forced vital capacity. Adverse effects, including influenza-like symptoms, were more common in the group receiving recombinant human DNase. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the routine use of mucolytics for bronchiectasis. High doses of bromhexine coupled with antibiotics may help with sputum production and clearance. PMID- 10796637 TI - Community interventions for preventing smoking in young people. AB - BACKGROUND: Decisions to smoke are made within a broad social context. Community interventions use co-ordinated, widespread, multi-component programmes to try and influence behaviour. OBJECTIVES: To determine the effectiveness of community interventions in preventing the uptake of smoking in young people. SEARCH STRATEGY: The Tobacco Addiction group specialised register, Medline and 21 other health, psychology and public policy electronic databases were searched, the bibliographies of identified studies were checked and contact was made with content area specialists. SELECTION CRITERIA: Randomised and non randomised controlled trials that assessed the effectiveness of multi-component community interventions compared to no intervention or to single component or school-based programmes only. Reported outcomes had to include smoking behaviour in young people under the age of 25 years. DATA COLLECTION AND ANALYSIS: Information relating to the characteristics and the content of community interventions, participants, outcomes and methods of the study was extracted by one reviewer and checked by a second. Studies were combined using qualitative narrative synthesis. MAIN RESULTS: Thirteen studies were included in the review, 44 studies did not meet all of the inclusion criteria. All studies used a controlled trial design, with four using random allocation of schools or communities. Of nine studies which compared community interventions to no intervention controls, two, which were part of cardiovascular disease prevention programmes, reported lower smoking prevalence. Of three studies comparing community interventions to school-based programmes only, one found differences in reported smoking prevalence. One study reported a lower rate of increase in prevalence in a community receiving a multi component intervention compared to a community exposed to a mass media campaign alone. One study reported a significant difference in smoking prevalence between a group receiving a media, school and homework intervention compared to a group receiving the media component only REVIEWER'S CONCLUSIONS: There is some limited support for the effectiveness of community interventions in helping prevent the uptake of smoking in young people. PMID- 10796638 TI - Individual behavioural counselling for smoking cessation. AB - BACKGROUND: Individual counselling from a smoking cessation specialist may help smokers to make a successful attempt to stop smoking. OBJECTIVES: The objective of the review is to determine the effects of individual counselling. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register for studies with counsel* in any field. Date of the most recent search: October 1998. SELECTION CRITERIA: Randomised or quasi-randomised trials with at least one treatment arm consisting of face to face individual counselling from a health care worker not involved in routine clinical care. The outcome was smoking cessation at follow-up at least six months after the start of counselling. DATA COLLECTION AND ANALYSIS: Both reviewers extracted data. The intervention and population, method of randomisation and completeness of follow-up were recorded. MAIN RESULTS: We identified eleven trials. Ten compared individual counselling to a minimal intervention, two compared two intensities of counselling, and one compared individual counselling to group therapy. Individual counselling was more effective than control. The odds ratio for successful smoking cessation was 1.55 (95% confidence interval 1.27 to 1.90). There was no evidence that more intensive counselling was more effective than brief counselling (odds ratio 1.17, 95% confidence interval 0.59 to 2.34). There was no evidence of a difference in effect between individual counselling and group therapy (odds ratio 1.33, 95% confidence interval 0.83 to 2.13). REVIEWER'S CONCLUSIONS: Smoking cessation counselling can assist smokers to quit. PMID- 10796639 TI - Aspirin for vascular dementia. AB - BACKGROUND: For patients with a diagnosis of vascular dementia there is evidence that aspirin is widely prescribed - in one study, completed by geriatricians and psychiatrists in the UK, 80% of patients with cognitive impairment (with vascular risk factors) were prescribed aspirin. However, a number of queries remain unanswered: Is there convincing evidence that aspirin benefits patients with vascular dementia? Does aspirin affect cognition or improve prognosis? In addition, does the risk of cerebral or gastric haemorrhage outweigh any benefit? The aim of this review is to assess the evidence of effectiveness of aspirin in those with a diagnosis of vascular dementia. OBJECTIVES: To assess the evidence of effectiveness of the use of aspirin for vascular dementia. SEARCH STRATEGY: Computerised databases were searched independently by two reviewers. In addition, relevant websites were searched and some journals were handsearched. Specialists in the field were approached for unpublished material and also any publications found were searched for additional references. SELECTION CRITERIA: All randomised controlled trials investigating the effect of aspirin for vascular dementia are included. Inclusion/exclusion of studies comprised systematic assessment of the quality of study design and the risk of bias. DATA COLLECTION AND ANALYSIS: Data were extracted independently by both reviewers, using a previously tested data extraction form and, where required, authors were contacted for data not provided in the papers. The aim was to evaluate data recorded via tools assessing cognitive and behavioural changes along with mortality, morbidity and institutionalisation data. MAIN RESULTS: One randomised controlled trial ( approximately approximately Meyer 1989 approximately approximately ) was included, and yielded data for analysis on a total of 70 patients. The only relevant outcome assessed in this trial was cognition. Change in cognitive outcome was towards being in favour of treatment. REVIEWER'S CONCLUSIONS: There is very limited evidence that aspirin is effective in treating patients with a diagnosis of vascular dementia. Further research is needed to assess the effect of aspirin on cognition, and also on additional outcomes such as behaviour, and quality of life. At present it is not possible to provide evidence for other queries regarding the use of aspirin for dementia (these are described in the Background section of this review). PMID- 10796640 TI - Olanzapine for schizophrenia. AB - BACKGROUND: Olanzapine is an atypical antipsychotic that is reported to be effective without producing the disabling extrapyramidal side effects associated with the older, typical antipsychotic drugs. OBJECTIVES: To determine the clinical effects and safety of olanzapine as compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and schizophreniform psychoses. SEARCH STRATEGY: The reviewers undertook electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 2, 1999), The Cochrane Schizophrenia Group's Register (September 1999), EMBASE (1980-1999), MEDLINE (1966-1999), and PsycLIT (1974-1999). References of all identified studies were searched for further trials, and the reviewers contacted relevant pharmaceutical companies and authors of trials. SELECTION CRITERIA: All randomised clinical trials comparing olanzapine to placebo or any antipsychotic treatment for those with schizophrenia or schizophreniform psychoses. DATA COLLECTION AND ANALYSIS: Data were independently extracted. For homogeneous dichotomous data the random effects relative risk (RR), the 95% confidence intervals (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data the reviewers calculated weighted mean differences. MAIN RESULTS: Twenty trials are included. Attrition from olanzapine versus placebo studies was so great (olanzapine - 61%, placebo - 73% by six weeks, RR 0.85 CI 0. 7-0.98, NNT 8 CI 5-40) that interpretation of results is problematic. Olanzapine appeared superior to placebo at six weeks for the outcome of 'no important clinical response' (RR 0.88 CI 0.8 0.98, NNT 8 CI 5-27), but trial data regarding negative symptoms are equivocal for this comparison. Dizziness and dry mouth were more common in the olanzapine treated group, and, although not statistically significant, the olanzapine group gained more weight. Data from several small trials are incomplete; but, for the short term outcome of 'no important clinical response', olanzapine seem as effective as typical antipsychotics (n=2778, RR 0.9 CI 0.76-1.06). Brief Psychiatric Rating Scale (BPRS) data tended to be equivocal but Positive and Negative Syndrome Scale (PANSS) rating of total score and negative and positive symptom sub-scores favoured olanzapine. With high attrition in both groups (olanzapine - 36%, typical drug - 49% by 6 weeks, n=2738, RR 0.85 CI 0.66-1.1; olanzapine - 83%, typical drug - 90% by 1 year, n=2738, RR 0.9 CI 0. 86-1.02), the assumptions included in all continuous data are considerable. Participants allocated olanzapine experienced fewer extrapyramidal side effects than people given haloperidol. Weight change data for the short term are not conclusive (n=2455, WMD 0.8kg CI -0.6-2.2) but the three to 12 month results suggest an average gain of four kilograms (n=233, WMD 4 CI 0.3-7.8). It is difficult to distinguish between olanzapine and other atypical drugs, although it may cause fewer extrapyramidal side effects than risperidone (n=339, RR 0.6 CI 0.4-0.9, NNH 8 CI 4-29). Olanzapine did cause more weight gain than its comparators but current data are not statistically significant (3-12 months, n=535, WMD 2.2kg CI 0.6-5). One study (n=180) found no clear differences between olanzapine and clozapine for people with treatment-resistant illness. REVIEWER'S CONCLUSIONS: For people with schizophrenia olanzapine may offer antipsychotic efficacy with fewer extrapyramidal side effects than typical drugs but more weight gain. The large proportions of participants leaving the studies early, in the large multi centre trials makes it difficult to draw firm conclusions on clinical effects. Large, long-term randomised trials with participants, interventions and primary outcomes that are familiar to those wishing to help those with schizophrenia are long overdue. PMID- 10796641 TI - Regimens of less than six months for treating tuberculosis. AB - BACKGROUND: WHO recommends 6 months of treatment in TB programmes. OBJECTIVES: The purpose of this review is to assess the effects of regimens lasting less than 6 months compared with longer regimens in the treatment of active TB. SEARCH STRATEGY: Search strategy: MEDLINE 1955-, Cochrane Infectious Diseases Trials Register, existing reviews, and researchers in the field. Date of the most recent search: January 1999. SELECTION CRITERIA: Randomized trials comparing two or more TB drug regimens, in which at least one regimen was <6 months and it was compared with at least one regimen that lasted longer, in any patients with active TB. DATA COLLECTION AND ANALYSIS: One reviewer extracted data and assessed trial quality. MAIN RESULTS: Seven trials with a total of 9 comparisons of <6 months (range: 2-5 months) versus longer treatment were included. About 2200 patients were in the shorter regimens and about 1900 in the longer regimens (the same comparison groups were used for more than one shorter regimen, in two studies). Relapse rates were consistently higher after shorter duration treatment regimens, regardless of the comparison made, though they were all relatively low. Results were significantly better in the longer groups in the meta-analyses of 2, 3, and 4 months of treatment vs longer treatment (Peto OR = 6.1 [95%CI 2.19,17.01], 3.67 [2.42,5.58], 3.64 [1.71,7. 75] but not in the single trial of 5 vs. 7 months (Peto OR = 2.24 [0. 90,5.59]. Relapse rates after longer (comparison) regimens ranged from 0-7% at one year (or more), and in the shorter treatment arms, they ranged from 1-9% in 8 trials, and18% relapsed in the one remaining. There was little or no difference in the rates of adverse reactions or toxicity requiring a change of regimen or discontinuation of treatment. The "sterilizing efficacy" at the end of treatment varied little among treatments, providing no predictive value for relapse rates. Few or no deaths were reported in the individual trials, and in no case did enough deaths occur for a comparison of short vs. long regimens. REVIEWER'S CONCLUSIONS: Longer periods of treatment (at least up to 6 months) result in higher success rates in patients with active TB, but the differences are small. Under field conditions, where adherence to treatment is a big problem, and shorter regimens might improve adherence, these differences may not be evident. A comparison of <6 months vs. 6 months of treatment under programme conditions would be needed to determine this. PMID- 10796642 TI - Isoniazid for preventing tuberculosis in non-HIV infected persons. AB - BACKGROUND: Although isoniazid (INH) is commonly used for treating tuberculosis (TB), it is also effective as preventive therapy. OBJECTIVES: The objective of this review was to estimate the effect of 6 and 12 month courses of INH for preventing TB in HIV-negative people at increased risk of developing active TB. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles. We hand-searched Science Citation Index and Index Medicus. SELECTION CRITERIA: Randomised trials of INH preventive therapy for 6 months or more compared with placebo. Follow-up for a minimum of 2 years. Trials enrolling patients with current or previously treated active TB, or with known HIV infection, were excluded. Criteria were applied by two reviewers independently. DATA COLLECTION AND ANALYSIS: Trial quality was assessed by two reviewers independently, and data extracted by one reviewer using a standardized extraction form. MAIN RESULTS: Eleven trials involving 73,375 patients were included. Trials were generally of high quality. Treatment with INH resulted in a relative risk (RR) of developing active TB of 0.40, (95% confidence interval ?CI? 0.31 to 0.52), over two years or longer. There was no significant difference between 6 and 12 month courses (RR of 0.44, 95% CI 0.27 to 0.73 for six months, and 0.38, 95% CI 0.28 to 0.50 for 12 months). Preventive therapy reduced deaths from TB, but this effect was not seen for all cause mortality. INH was associated with hepatotoxicity in 0.36% of people on 6 months treatment and in 0.52% of people treated for 12 months. REVIEWER'S CONCLUSIONS: Isoniazid is effective for the prevention of active TB in diverse at-risk patients, and six and 12 month regimens have a similar effect. PMID- 10796643 TI - Zinc for the common cold. AB - OBJECTIVES: Interest in zinc as a treatment for the common cold has grown following the recent publication of several controlled trials. The objective of this review was to assess the effects of zinc lozenges for cold symptoms. SEARCH STRATEGY: A search was made of the Cochrane Controlled Trials Register, MEDLINE, EMBASE and reference lists of articles. Searches were run to the end of 1997. SELECTION CRITERIA: Randomised double blind placebo-controlled trials of zinc for acute upper respiratory tract infection or cold. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Seven trials involving 754 cases were included. With the exception of one study, the methodological quality was rated as medium to high. For most outcome measures different summary estimates were used across the studies to describe the duration, incidence and severity of respiratory symptoms. This limited the ability to pool results. Results from two trials (04 - Mossad; 08 - Smith) suggested zinc lozenges reduced the severity and duration of cold symptoms. However, there was significant potential for bias, and further research is required to substantiate these findings. Overall, the results suggest that treatment with zinc lozenges did not reduce the duration of cold symptoms. REVIEWER'S CONCLUSIONS: Evidence of the effects of zinc lozenges for treating the common cold is inconclusive. Given the potential for treatment to produce side effects, the use of zinc lozenges to treat cold symptoms deserves further study. PMID- 10796644 TI - Validation therapy for dementia. AB - BACKGROUND: Validation therapy is a "therapy for communicating with old-old people who are diagnosed as having Alzheimer's disease and related dementia". The approach was developed by Naomi Feil between 1963 and 1980. She classifies individuals with cognitive impairment as having one of four stages on a continuum of dementia: these stages are Malorientation, Time Confusion, Repetitive Motion and Vegetation. The benefits of Validation therapy for patients are reported by Feil as restoration of self worth, minimisation of the the degree to which patients withdraw from the outside world, promotion of communication and interaction with other people, reduction of stress and anxiety, stimulation of dormant potential, help in resolving unfinished life tasks, facilitation of independent living for as long as possible. These benefits are highly desirable and Validation therapy but there is a need to show them on the basis of substantive research yielding strong evidence across a number of well designed studies. The potential number of people that might benefit from the implementation of Validation therapy is considerable given the increasing incidence of dementia. OBJECTIVES: To evaluate the effectiveness of using validation therapy with people diagnosed as having senile dementia of the Alzheimer's type, other forms of dementia, or cognitive impairment. SEARCH STRATEGY: The Cochrane Controlled Trials Register was searched by the Cochrane Dementia and Cognitive Impairment Group Coordinator and relevant references were forwarded to the primary reviewer. The reviewers carried out independent searches of the following electronic databases - MEDLINE, EMBASE, Eureka, CINAHL, PSYCLIT, RLIN, SIGLE and SOCIOFILE. The following key terms were used in searches; validation-therapy, dementia, cognitive impairmnent, communication, and random* control*. An author search using Feil and Naomi was also carried out. Personal contact was made with the Validation Institute, Cleveland, Ohio USA and unpublished studies were sought by direct contact with the relevant authors, or institutions. Key individuals who might provide further information regarding validation therapy were contacted. SELECTION CRITERIA: All randomised controlled trials (RCTs) and quasi-randomised controlled trials examining Validation therapy as an intervention for dementia were considered for inclusion in review. The criteria for inclusion/exclusion comprised systematic assessment of the quality of study design and the risk of bias, using a standard data extraction form. Outcome measures of cognitive and/or behavioural change were required. DATA COLLECTION AND ANALYSIS: Data were extracted independently by both reviewers, using a previously tested data extraction form. Authors were contacted for data not provided in the papers. Psychological scales measuring cognitive and behavioural changes were examined. Subcategory analysis was performed due to the diversity of the scales and the inability to combine data across studies. MAIN RESULTS: Only three studies were identifed that potentially met the inclusion criteria and only two small studies were able to be obtained. The Peoples 1982 study was not obtainable through a British Library loan because the cost was prohibitive. Data from 2 studies were entered using MetaView - Robb 1986, Toseland et al 1997, incorporating data on a total of 87 patients (32 in the experimental group, and 55 in the control groups (usual care 34 and social contact 21)). It was not possible to pool the data from the two included studies, and therefore, subcategory analyses were carried out. The analysis of the data failed to reveal statistically significant results although there were trends toward favouring validation therapy for some outcomes. (ABSTRACT TRUNCATED) PMID- 10796645 TI - Iron chelating agents for treating malaria. AB - BACKGROUND: Mortality from Plasmodium falciparum malaria remains high; death and sequelae occur in even in patients treated with antimalarial drugs. Researchers are exploring the effects of adding treatments to the main antimalarial regimens in an attempt to reduce mortality. Iron chelation is one potential chemotherapeutic adjuvant treatment. Before advocating adjunctive therapy, the effects of iron chelators in improving patient outcomes needs to be examined. OBJECTIVES: To assess the effects of iron-chelating agents combined with antimalarial drugs, or iron chelators alone, for treating Plasmodium falciparum malaria in adults and children, in relation to mortality, coma recovery time, parasite clearance, and adverse effects. SEARCH STRATEGY: Electronic searches of the Cochrane Library, MEDLINE, and EMBASE, using the standard Cochrane search strategy. Bibliographies of retrieved studies were scrutinized in order to identify further relevant trials. Organisations, experts and other individuals in malaria research were contacted for unpublished studies. SELECTION CRITERIA: All randomised controlled trials of adults or children with P.falciparum malaria. DATA COLLECTION AND ANALYSIS: Trials were identified and extracted by a single reviewer (HS) and checked by a second (MM). Inclusion criteria were applied, and data were extracted independently by both reviewers. Authors were contacted for missing and additional data. Meta-analysis used Relative Risk (RR) and 95% Confidence Intervals. MAIN RESULTS: No evidence of benefit or harm were shown in relation to mortality, but studies were small, and one trial was tending towards more deaths with the intervention when it was stopped. The risk of experiencing persistent seizures was significantly lower with desferrioxamine compared to placebo treatment (RR 0.80, 95% CI 0.67 to 0.95). Many adverse effects were more common in participants treated with desferrioxamine. REVIEWER'S CONCLUSIONS: Trends suggestive of both harm (death) and potential benefit (fewer seizures) are demonstrated in this review. It is not possible to comment on time to event outcomes that include coma recovery or parasitaemia as we are clarifying data with the trialists. Whether to conduct further trials will depend on a judgement about potential benefit. PMID- 10796646 TI - Blood transfusion for treating malarial anaemia. AB - BACKGROUND: Blood transfusion is used in patients with severe malarial anaemia, but risks adverse reactions, transmission of disease, and is complicated to organise in developing countries. OBJECTIVES: This review evaluates the effects of routine blood transfusion for severe anaemia on death and adverse outcomes in malarious areas. SEARCH STRATEGY: The Cochrane Controlled Trials Register, African Index Medicus, LILACS, EMBASE and reference lists of relevant articles, and contact with researchers and organizations working in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of blood transfusion compared with conservative management in malaria-associated severe anaemia. DATA COLLECTION AND ANALYSIS: Trials were identified and extracted by a single reviewer (MM) and checked by a second (HS). Inclusion criteria were applied and data were extracted independendtly by both reviewers. MAIN RESULTS: Two randomised trials of 230 children were included. In the transfusion group, there was a non-significant tendency towards fewer deaths (RR 0.41, 95% CI 0.06 to 2.70), but significantly more severe adverse events (RR 8.60, 95% CI 1.11 to 66. 43). In one trial by Bojang (1997a) respiratory distress was less common and hospital stay was shorter in the transfusion group (WMD 1.9 days, 95% CI 2.4 to 1.3). Subsequent need for urgent blood transfusion was less common in the transfusion group (RR 0.11, 95% CI 0.02 to 0.62). Day 28 packed cell volume was less in the transfusion group (WMD -1.34, 95% CI -2.57 to -0.11). There was no information on HIV or Hepatitis B virus transmission. REVIEWER'S CONCLUSIONS: There is insufficient data to be sure whether routinely giving blood to clinically stable children with severe anaemia in endemic malarious areas reduces death, or results in higher haematocrit measured at one month. PMID- 10796647 TI - Antibiotics for preventing pneumonia in children with measles. AB - BACKGROUND: Measles causes more than a million deaths a year, of which most are children under five years of age who die from pneumonia. OBJECTIVES: The objective of this review was to assess the effects of antibiotics given to children with measles on reducing pneumonia or mortality, and to assess whether antibiotics should be given to all children with measles in communities with a high fatality rate. SEARCH STRATEGY: We searched MEDLINE (1966 - 1999), EMBASE (1980-1999) and the specialized trials register of the Acute Respiratory Infections Group in August 1999, and all relevant journals in the University of Melbourne medical library for the years 1935-46. SELECTION CRITERIA: Randomised or controlled trials of antibiotics for children with measles. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Six trials with 1304 children were included. All but one of the trials were unblinded, and randomisation was either not described or was by alternate allocation. In four studies, the incidence of pneumonia in the control group was similar to that in the antibiotic prophylaxis group; in the other two studies, the incidence of pneumonia was unusually high in the control group so these children had a higher complication rate than the antibiotic group. Four of the 764 children given antibiotics died compared with one of the 637 controls. REVIEWER'S CONCLUSIONS: The quality of the trials reviewed was poor, and they provide very weak evidence for giving antibiotics to all children with measles. Available evidence suggests that antibiotics should be given only if a child has clinical signs of pneumonia or other evidence of sepsis. PMID- 10796648 TI - Acellular vaccines for preventing whooping cough in children. AB - OBJECTIVES: Routine use of whole cell pertussis vaccines was suspended in some countries in the late 1970s and early 1980s, leading to a resurgence of whooping cough. Acellular pertussis vaccines containing purified or recombinant Bordetella pertussis antigens were developed in the hope that they would be as effective but less toxic than the whole cell vaccines. The objective of this review was to assess the effects of acellular pertussis vaccines in children. SEARCH STRATEGY: The Cochrane Controlled Trials Register and Medline were searched up to January 1998. SELECTION CRITERIA: Double-blind randomised efficacy and safety trials of acellular pertussis vaccines in children, with active follow-up of participants and laboratory verification of pertussis cases. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Six efficacy trials and 45 safety trials were included. Acellular pertussis vaccines with three or more pertussis vaccines were more effective than those with one or two antigens. They were also more effective than one type of whole cell pertussis vaccine, but less effective than two other types of whole cell vaccines. Differences in trial design precluded pooling of the efficacy data and results should be interpreted with caution. Most systemic and local adverse events were significantly less common with acellular than with whole cell pertussis vaccines. REVIEWER'S CONCLUSIONS: Multi-component acellular pertussis vaccines are effective, and show less adverse effects than whole cell pertussis vaccines. However in areas where whooping cough is more likely to be fatal, the higher toxicity of some whole cell vaccines may be offset by their increased effectiveness. PMID- 10796649 TI - Graft type for femoro-popliteal bypass surgery. AB - BACKGROUND: Vascular surgeons construct femoro-popliteal bypass grafts, from the groin to the knee, to save limbs that might otherwise require amputation in patients with severe arterial disease, and to improve walking distance in patients with less severe arterial disease. During the operation, the blocked native artery is bypassed using either a section of the patient's own vein (autologous vein), human umbilical vein (HUV), or an artificial graft e.g. Dacron or polytetrafluoroethylene (PTFE). OBJECTIVES: The objective of this review was to determine the most effective type of graft for femoro-popliteal bypass surgery. SEARCH STRATEGY: The reviewers searched the Cochrane Peripheral Vascular Diseases Group trials register, reference lists of relevant articles, and hand searched proceedings from the British and European Vascular Surgical Societies and the North American Society of Vascular Surgery. They also contacted all major manufacturers of artificial grafts and authors of published trials to enquire about unpublished trials. SELECTION CRITERIA: Randomised trials comparing one type of femoro-popliteal graft with another. DATA COLLECTION AND ANALYSIS: Both reviewers selected trials and assessed trial quality independently. MAIN RESULTS: Nine trials were included with a total of 1334 participants. These investigated a variety of graft types. In one trial of above-knee grafting, primary and secondary patency were significantly better for saphenous vein (73% and 90%, respectively) compared to PTFE (47%, p<0.05 and 47%, p<0.05) and Dacron (54%, p<0.01 and 60%, p<0.01) at four years. Two trials comparing in-situ and reversed saphenous vein grafts to the above- and below-knee popliteal artery revealed no differences in primary patency (64% v 62% respectively), secondary patency (65% v 70%), or survival with intact limb (74% both groups) with five to ten year follow up. Three trials comparing PTFE with HUV showed significantly better secondary patency rates for HUV, (41% v 73%, p<0.005; 49% v 66%, p<0.05; 22% v 42%,p=0.005) one also showed significantly better primary patency for HUV at five years (32% v 65%, p<0.001). Comparison of PTFE grafts with, and without, a vein cuff found no difference in above-knee grafts. However, primary patency below-knee was higher with a PTFE plus vein cuff bypass (52% v 29%, p=0.03) at two years. REVIEWER'S CONCLUSIONS: There is no clear evidence which type of graft is best for femoro popliteal grafting. In terms of autologous graft patency, in-situ and reversed vein grafts are equally successful, while HUV performs better than PTFE. A distal vein cuff may improve primary patency for below-knee PTFE femoro-popliteal grafts. PMID- 10796650 TI - Magnesium sulfate for treating exacerbations of acute asthma in the emergency department. AB - BACKGROUND: Treatment of acute asthma is based on rapid reversal of bronchospasm and arresting airway inflammation. There is some evidence that intravenous magnesium can provide additional bronchodilation when given in conjunction with standard bronchodilating agents and corticosteroids. No systematic review of this literature has been completed on this topic. OBJECTIVES: To examine the effect of additional intravenous magnesium sulfate in patients with acute asthma managed in the emergency department. SEARCH STRATEGY: Randomised controlled trials were identified from the Cochrane Airways Review Group register. Bibliographies from included studies, known reviews and texts were searched. Primary authors and content experts were contacted. SELECTION CRITERIA: Randomised controlled trials or quasi-randomised trials were eligible for inclusion. Studies were included if patients presented with acute asthma and were treated with IV magnesium sulfate vs placebo. DATA COLLECTION AND ANALYSIS: Data were extracted and methodological quality was assessed independently by two reviewers. Missing data were obtained from authors. MAIN RESULTS: Seven trials were included (5 adult, 2 pediatric). A total of 665 patients were involved. Patients receiving magnesium sulfate demonstrated non-significant improvements in peak expiratory flow rates when all studies were pooled (weighted mean difference: 29.4 L/min; 95% confidence interval: -3.4 to 62). In studies of people with severe acute asthma, peak expiratory flow rate improved by 52.3 L/min (95% confidence interval: 27 to 77.5). The forced expiratory volume in one second also improved by 9.8 % predicted (95% confidence interval: 3.8 to 15.8). Overall, admission to hospital was not reduced, odds ratio: 0.31 (95% confidence interval: 0.09 to 1.02). In the severe subgroup, admissions were reduced in those receiving magnesium sulfate (odds ratio: 0.10, 95% confidence interval: 0.04 to 0.27). No clinically important changes in vital signs or adverse side effects were reported. REVIEWER'S CONCLUSIONS: Current evidence does not support routine use of intravenous magnesium sulfate in all patients with acute asthma presenting to the emergency department. Magnesium sulfate appears to be safe and beneficial in patients who present with severe acute asthma. PMID- 10796651 TI - Inhaled beta-agonists for asthma in mechanically ventilated patients. AB - BACKGROUND: A small number of patients with acute severe asthma require intubation and positive pressure ventilation. The beneficial effects of inhaled bronchodilators on acute asthma in spontaneously breathing subjects are well established, but there remain important questions regarding inhaled beta2 agonists, for patients who are intubated and receiving ventilation. OBJECTIVES: To determine the effects of inhaled beta-agonists on asthmatic patients who require intubation and mechanical ventilation. SEARCH STRATEGY: Randomised controlled trials were sought from the Cochrane Airways Group Asthma Register. Primary authors and content experts were contacted to identify eligible studies and bibliographies from known reviews and texts were searched. SELECTION CRITERIA: Randomised, controlled clinical trials involving adult patients with acute asthma, who were intubated and supported with positive pressure ventilation. Studies were to be included if patients were treated with beta2 adrenergic agonist agents and there was a comparator group treated with either placebo, no medication, or 'standard' treatment. DATA COLLECTION AND ANALYSIS: Two reviewers independently examined all identified articles. The full text of any potentially relevant article was reviewed independently by two reviewers. MAIN RESULTS: The search yielded 152 abstracts. Of these, four articles were identified as potential trials. None of the four trials met the inclusion criteria for the review. REVIEWER'S CONCLUSIONS: There are no data from randomised controlled trials to provide evidence for or against current practices regarding the use of inhaled beta2-agonists in asthmatic subjects who are intubated and ventilated. PMID- 10796652 TI - Short-acting beta 2 agonists for stable chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is a chronic condition characterised by progressive airflow limitation that is at most partially reversible. Despite the lack of reversibility patients often report symptomatic improvement with short-acting beta 2 bronchodilator medications. They are used on either an "as required" or a "regular plus as required basis" and they may be used in conjunction with other bronchodilator medicines such as ipratropium and methylxanthines. These medicines are used in the management of both stable and acute exacerbations of COPD. This review examined the effect of short-acting beta 2 bronchodilators given by inhalation in stable COPD. OBJECTIVES: To determine the clinical effect and assess the adverse effects of inhaled short-acting beta 2 agonist bronchodilators compared with placebo in stable COPD. SEARCH STRATEGY: Only randomised controlled trials (RCTs) were considered. RCTs were identified using the Cochrane Airways Group database (CENTRAL). In addition, the reference lists of review articles and RCTs retrieved in full were searched for other potentially relevant citations. SELECTION CRITERIA: Only trials with adult patients with stable COPD, as defined by internationally accepted guidelines (ATS, ERS or BTS) were included in this review. All trials had a minimum duration of 7 days of regular treatment with short-acting beta 2 bronchodilators given by inhalation and compared with placebo. Data from trials where beta 2 agonists were used alone or in combination with other medicines (e.g. ipratropium bromide) were used only if there was a direct comparison between beta 2 bronchodilator alone and placebo. DATA COLLECTION AND ANALYSIS: Outcomes were analysed as continuous or dichotomous outcomes, using standard statistical techniques. For continuous outcomes, the weighted mean difference (WMD) and 95% confidence intervals were calculated and for dichotomous outcomes, the odds ratio was calculated with 95% confidence intervals by Peto's methods. Funnel plots were used to test for publication bias. MAIN RESULTS: Thirteen studies were included in this review. Most had small sample sizes and some of the sutides used very short-acting outdated compounds. All the studies used a cross-over design and were of high quality. Spirometry done at the end of study period was measured after administration of treatment (post-bronchodilator) which showed both FEV1 (0.150 L/min, 95%CI: 0. 02-0.28) and FVC (0.310 L, 95%CI: 0.00-0.62) to improve significanly but slightly when compared to placebo. A few studies measured FRC, airway resistance or conductance at the end of the study period. In all cases these measurements were done several hours after treatment, and no significant differences (p>0.05 in all cases) were found between the bronchodilator and placebo groups. Walking test Large increases in 6MW distance was observed in two studies, however one study did not show any positive improvements. There was a large increase in the 12MW distance as shown by one study. Due to the small number of studies reporting this outcome no significant differences were found in the walking distance between the bronchodilator and placebo groups. Peak Flow Rate Both morning (36. 04 L/min; 95%CI: 0.80-71.27) and evening (36.68 L/min; 95%CI: 2. 47-70.89) PEFR were significantly higher during active treatment than during placebo. Symptoms Breatlessness was measured on various scales therefore data that were presented in a suitable form were combined using standardized means for inclusion in the analysis. A significant improvement (-0.33; 95%CI: 0.58 to -0.07 with p=0.01) in the breathlessness score was observed during treatment with beta-2 agonist when compared to placebo. Cough was reported to improve significantly (data not usable) during treatment with beta2 agonist in one study but not in two others. A non-significant decrease in sputum production was reported by Wilson 1980, however four other studies reported no PMID- 10796653 TI - Gastro-oesophageal reflux treatment for asthma in adults and children. AB - BACKGROUND: Asthma and gastro-oesophageal reflux are both common medical conditions and often co-exist. Studies have shown conflicting results concerning the effects of lower oesophageal acidification as a trigger of asthma. Furthermore, asthma might precipitate gastro-oesophageal reflux. Thus a temporal association between the two does not establish that gastro-oesophageal reflux triggers asthma. Randomised trials of a number of treatments for gastro oesophageal reflux in asthma have been conducted, with conflicting results. OBJECTIVES: The objective of this review was to evaluate the effectiveness of treatments for gastro-oesophageal reflux in terms of their benefit on asthma. SEARCH STRATEGY: The Cochrane Airways Group trials register, review articles and reference lists of articles were searched. SELECTION CRITERIA: Randomised controlled trials of treatment for oesophageal reflux in adults and children with a diagnosis of both asthma and gastro-oesophageal reflux. DATA COLLECTION AND ANALYSIS: Trial quality and data extraction were carried out by two independent reviewers. Authors were contacted for confirmation or more data. MAIN RESULTS: Nine trials met the inclusion criteria. Interventions included proton pump inhibitors (n=3), histamine antagonists (n=5), surgery (n=1) and conservative management (n=1). Treatment duration ranged from 1 week to 6 months. A temporal association between asthma and gastro-oesophageal reflux was investigated in 4 trials and found to be present in a proportion of participants in these trials. Anti-reflux treatment did not consistently improve lung function, asthma symptoms, nocturnal asthma or the use of asthma medications. REVIEWER'S CONCLUSIONS: In asthmatic subjects with gastro-oesophageal reflux, (but who were not recruited specifically on the basis of reflux-associated respiratory symptoms), there was no overall improvement in asthma following treatment for gastro-oesophageal reflux. Subgroups of patients may gain benefit, but it appears difficult to predict responders. PMID- 10796654 TI - Interventions for preventing tobacco sales to minors. AB - BACKGROUND: Laws restricting sales of tobacco products to minors exist in many countries, but young people may still purchase cigarettes easily. OBJECTIVES: The review assesses the effects of interventions to reduce underage access to tobacco by deterring shopkeepers from making illegal sales. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction group trials register and Medline. Date of the most recent searches: July 1999. SELECTION CRITERIA: We included controlled trials and uncontrolled studies with pre- and post intervention assessment of interventions to change retailers' behaviour. The outcomes were changes in retailer compliance with legislation (assessed by test purchasing), changes in young people's smoking behaviour, and perceived ease of access to tobacco products. DATA COLLECTION AND ANALYSIS: Studies were prescreened for relevance by one person and assessed for inclusion by two people independently. Data from included studies were extracted by one person and checked by a second. Study designs and types of intervention were heterogeneous so results were synthesised narratively, with greater weight given to controlled studies. MAIN RESULTS: We identified 27 studies of which 13 were controlled. Giving retailers information was less effective in reducing illegal sales than active enforcement and/or multicomponent educational strategies. No strategy achieved complete, sustained compliance. In three controlled trials, there was little effect of intervention on youth perceptions of access or prevalence of smoking. REVIEWER'S CONCLUSIONS: Interventions with retailers can lead to large decreases in the number of outlets selling tobacco to youths. However, few of the communities studied in this review achieved sustained levels of high compliance. This may explain why there is limited evidence for an effect of intervention on youth perception of ease of access to tobacco, and on smoking behaviour. PMID- 10796655 TI - Thiamine for Alzheimer's disease. AB - BACKGROUND: Vitamin B1 (thiamine) plays an important role in Wernicke-Korsakoff syndrome (a form of amnesia caused by brain damage occurring in long-term alcoholics who rely mainly on alcohol for nutrition). The acute syndrome is normally reversible but may proceed to profound dementia, although its progress can be stopped by a timely injection of a large dose of thiamine. There have been suggestions that thiamine may have a beneficial effect in Alzheimer's disease. OBJECTIVES: The objective of this systematic review is to evaluate evidence of the effect of thiamine for Alzheimer's disease. SEARCH STRATEGY: The Cochrane Controlled Trials Register and the CDCIG Register were searched using the terms 'thiamine*, alzheimer* and vitamin* B1'. Other sources were also searched. SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with thiamine was administered for more than a day and compared to placebo in patients with dementia of the Alzheimer's type. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers, pooled if appropriate and possible, and the pooled odds ratios (95% CI) or the average differences (95% CI) were estimated. No intention-to-treat data were available to be used. MAIN RESULTS: Few data were available for review. The data were compatible with thiamine producing harm, no change or improvement. For measures of cognition, the effect of thiamine was non-significantly worse than placebo on the Mini Mental State Examination score (0-30; high=good) at 12 months: WMD -4.3 (95% CI: -14.4 - +5.8) and at time points 3, 6 and 9 months. Change from baseline analyses showed placebo to be significantly better than thiamine at all time points beyond three months; WMD -4.8 (95% CI: -6.0 to -3.6) at 12 months. There was no statistically significant difference in the test of Verbal Fluency and the Boston Naming Test. These analyses were based only on those who completed the study and not on intention-to-treat analyses. There were no results presented for withdrawal by treatment group. Data on measures of functional autonomy, behaviour, quality of life, dependency, or effect on carer were not available. REVIEWER'S CONCLUSIONS: This review finds no evidence that thiamine is a useful treatment for the symptoms of Alzheimer's disease. The data are so poor and sparse that it is difficult to state almost anything of its effect in Alzheimer's disease. Thiamine cannot be recommended for patients with Alzheimer's disease. PMID- 10796656 TI - Patient held clinical information for people with psychotic illnesses. AB - BACKGROUND: It is important to seek cost-effective methods of improving the care and outcome of those with serious mental illnesses. Patient-held records, where the person with the illness holds all or some personal information relating to the course and care of their illness, are now the norm in some clinical settings. Their value for those with serious mental illnesses is unknown. OBJECTIVES: To evaluate the effects of personalised and accessible patient-held clinical information for people with a diagnosis of psychotic illness. SEARCH STRATEGY: Electronic searches of AMED (1980-1998), Biological Abstracts (1985-1998), British Nursing Index (1994-1998), CAB (1973-1999), CINAHL (1982-1999), The Cochrane Controlled Trials Register (Issue 1, 1999), EMBASE (1980-1999), HEALTHSTAR (1990-1999), HMIC (King's Fund Database 1979-1998 & HELMIS 1984-1998), MEDLINE (1966-1999), PsycLIT (1887-1999), Royal College of Nursing Database (1985 1996), SIGLE (1990-1998), Sociological Abstracts (1963-1998) and the Internet (http://www. controlled-trials.com/) were undertaken. This was supplemented by personal contact with the Executive Board of the European Network for Mental Health Service Evaluation. SELECTION CRITERIA: The inclusion criteria were that studies should: i. be randomised or quasi-randomised trials; ii. have involved adults with a diagnosis of a psychotic illness; and iii. compare any personalised and accessible clinical information held by the patient beyond standard care to standard information routinely held such as appointment cards and generic information on diagnosis, treatment or services available. DATA COLLECTION AND ANALYSIS: Study selection and data extraction was reliability undertaken. Analysis was not possible. MAIN RESULTS: Not one study met the inclusion criteria for the review. One study (Stafford 1997) was found on the use of client held records for people with long term mental illness but the participants had not been randomised. Two important randomised studies (Lester 1999, Papageorgiou 1999) are ongoing. REVIEWER'S CONCLUSIONS: There is a gap in the evidence regarding patient-held, personalised, accessible clinical information for people with psychotic illnesses. It cannot be assumed that patient-held information is beneficial or cost-effective without evidence from well planned, conducted and reported randomised trials. PMID- 10796657 TI - Sertindole for schizophrenia. AB - BACKGROUND: Sertindole is an atypical antipsychotic, which is thought to give a lower incidence of extrapyramidal side effects at clinically effective doses than typical antipsychotic drugs. In December 1998, Lundbeck Ltd., the manufacturers of sertindole, voluntarily suspended the availability of the drug due to concerns about cardiac arrhythmia and sudden cardiac death associated with its use. Sertindole has therefore been withdrawn from the market pending discussion with the European Regulatory Authority over cardiac safety. OBJECTIVES: To determine the effects of sertindole compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), EMBASE (1980-1999), LILACS (1982-1996), MEDLINE (1966-1999), PSYNDEX (1977-1995) and PsycLIT (1974-1999) were undertaken. In addition, pharmaceutical databases on the Dialog Corporation Datastar and Dialog services were searched. References of all identified studies were searched for further trials. The manufacturer of sertindole and authors of trials were contacted. SELECTION CRITERIA: All randomised controlled trials that compared sertindole to placebo or other antipsychotic drug treatments were included by independent assessment. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. For homogeneous dichotomous data the risk ratio (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) or numbers needed to harm (NNH) were calculated on an intention-to-treat basis. For continuous data, weighted mean differences (WMD) were calculated. All data were inspected for heterogeneity. MAIN RESULTS: Two large important studies were excluded, because they did not report any usable data. The two that were included suggested that sertindole was more antipsychotic than placebo, as acceptable as placebo and better tolerated than haloperidol (NNT=9, RR 0.63 CI 0.41 to 0.96). Sertindole was associated with fewer movement disorders than haloperidol but was shown to cause more weight gain (NNH=9 RR 6.33, CI 1.92 to 20.92), rhinitis (NNH=8, RR 1.74, CI 1,28 to 2.36) and possibly male sexual dysfunction. Cardiac problems (QTc intervals of at least 500msec) were evident even in the randomised trials (NNH=13 RR 23, CI 1.37 to 386.60). REVIEWER'S CONCLUSIONS: Because of the cardiac problems, even evident within poorly reported studies, at present sertindole should, if possible, be avoided. If sertindole is to be reintroduced, gold-standard evidence of its clinical benefits will need to far outweigh its real risks. PMID- 10796658 TI - Immunoglobulin for preventing respiratory syncytial virus infection. AB - BACKGROUND: Respiratory Syncytial virus, the most important cause of lower respiratory tract infections in infants and young children in industrialized countries, is associated with increased morbidity in premature infants with or without bronchopulmonary dysplasia as well as those with congenital heart disease. Because of observations that lower rates of disease occur immediately after birth, presumably due to vertical transmission of maternal antibody, and animal studies where protection from pneumonia was observed through administration of immune globulin, the efficacy of passive prophylaxis in premature infants has been studied. OBJECTIVES: This meta-analysis was performed to assess the effects of polyclonal respiratory syncytial virus hyperimmune globulin or monoclonal antibody in preventing RSV hospitalization, receipt of intensive care, mechanical ventilation, and mortality in those with underlying prematurity, bronchopulmonary dysplasia, or congenital heart disease. SEARCH STRATEGY: We searched the Cochrane Acute Respiratory Infections trials register and MEDLINE in March, 1999. In addition, abstracts on these topics were sought from the Pediatric Academies Meetings and the Intersciences Conference on Antimicrobial Agents and Chemotherapy for the years 1994 to 1997, inclusive. SELECTION CRITERIA: Randomized, controlled trials of prevention of RSV using immune globulin, respiratory syncytial virus immune globulin, or monoclonal RSV antibody in children with prematurity, bronchopulmonary dysplasia or congenital heart disease. DATA COLLECTION AND ANALYSIS: Two authors independently abstracted data and assessed study quality. Only incidences could be pooled because data for durations was summarized in a manner that did not allow combining results across the studies. MAIN RESULTS: Four studies with a total of 2598 subjects were included in the main analysis. All were randomized controlled trials. Two trials were not blinded. Three studies examined RSV hyperimmune globulin and one examined monoclonal RSV antibody. A study of a different monoclonal RSV antibody could not be included because it has not been presented or published. The pooled Peto Odds Ratios favoring prophylaxis were 0.48 (95% CI 0.37, 0.64), 0.47 (0.29, 0. 77), and 0.99 (0.48, 2.07) for incidence of hospitalization, incidence of ICU admission, and incidence of mechanical ventilation, respectively. The numbers needed to prevent one hospitalization and one ICU admission are 17 and 50 respectively. REVIEWER'S CONCLUSIONS: RSVIG is effective in preventing RSV hospitalizations and admission to the intensive care unit, but not in preventing mechanical ventilation. There was a non-significant trend towards a higher mortality in children given RSVIG. PMID- 10796659 TI - Heated, humidified air for the common cold. AB - OBJECTIVES: To assess the effects of inhaling heated water vapour with the help of a rhinotherm (an equipment designed to deliver heated water vapour to a person's nasal cavity), in the treatment of the common cold by comparing a. symptoms b. viral shedding c. nasal resistance after a natural or experimentally induced common cold. SEARCH STRATEGY: We searched MEDLINE with MeSH headings: common cold, rhinopharyngitis, inhalation, steam, heated vapour, rhinothermy, till July 1999. EMBASE, Current Contents, review articles, cross references were also searched. Attempts were also made to contact the manufacturers for any unpublished data. SELECTION CRITERIA: Randomized trials using heated water vapour in a standardized way in patients with the common cold or volunteers with experimental induction of rhinovirus infection were included in the review. DATA COLLECTION AND ANALYSIS: All the articles retrieved were initially subjected to a review for inclusion / exclusion criteria. Review articles, editorials, abstracts with inadequate outcome description were excluded. Studies selected for inclusion were subjected to a methodological assessment. MAIN RESULTS: The results of a systematic review of six trials with 319 participants, support the use of warm vapour inhalations in the common cold in terms of relief of symptoms (Odds Ratio with 95 % CI 0.31, 0.16-0.60, Relative risk 0.56, 0.4-0. 79). Results on symptom score indices were equivocal. None of the studies demonstrated a worsening of clinical symptom scores. One study demonstrated increased nasal resistance one week after steam inhalation in contrast to an earlier study which showed improvement in the nasal resistance. There was no evidence of decreased viral shedding measured by virus isolation in the nasal secretions (Tyrrell 1989) or measurement of viral titres in nasal washings among treatment group. The rhinovirus titres in the nasal washings from the treatment group were the same as those of the placebo group on day one prior to the treatment and on all four days after the treatment. The area under curve (Hendley 1994) was also similar in the placebo and treatment groups for titres of virus in the nasal washings as were the average viral titres across five days of follow up, the maximum values after treatment, and viral shedding velocity i.e. amount of virus shed per day (Hendley 1994). Minor side effects due to thermal stress were reported in all the studies. REVIEWER'S CONCLUSIONS: Three trials demonstrated beneficial effects on the symptoms of the common cold. One study from Israel showed a decrease in nasal resistance measured by peak nasal expiratory and inspiratory flow rate. Studies done in North America failed to show any objective improvement in outcome measures with the study intervention. A multi-centre double blind randomised controlled trial testing this therapy with uniform outcome measures is recommended. PMID- 10796660 TI - Medical day hospital care for the elderly versus alternative forms of care. AB - BACKGROUND: Geriatric day hospitals provide multi-disciplinary rehabilitation in an outpatient setting. Concern has been expressed that evidence for effectiveness is equivocal and that day hospital care is expensive. OBJECTIVES: To assess the effects of medical day hospitals for elderly people. SEARCH STRATEGY: We searched The Cochrane Library, MEDLINE, Sigle, Bids, and Cinahl up to January 1997, and reference lists of articles. We also searched Index Medicus and International Dissertation Abstracts up to January 1997. We contacted authors of previous studies of day hospital care. SELECTION CRITERIA: Randomised trials comparing geriatric medical day hospitals with alternative forms of care. The participants were elderly medical patients. The outcomes were death, place of residence, dependency, global 'poor' outcome (death, institutionalisation or dependency), activities of daily (ADL) score, subjective health status, patient satisfaction, and resource use. DATA COLLECTION AND ANALYSIS: Three reviewers independently extracted data and assessed study quality. MAIN RESULTS: Twelve studies were included involving 22 day hospitals and 2867 patients. Five studies compared day hospital with comprehensive elderly care, four compared day hospital with domiciliary care and three compared day hospital with no comprehensive elderly care. There were no significant differences between day hospital attendance and comparison treatments for the outcomes of death, death or requiring institutional care, death or deterioration in ADL. When death or a 'poor' outcome at follow up was examined there was a significant difference in favour of day hospital attendance when compared to no comprehensive elderly care (odds ratio 0.73; 95% confidence interval 0.53-1.00; P < 0.05). Dependency was measured in 11 studies using a variety of ADL measures; two described short-term improvement for the day hospital group, one reported improved outcome for the comparison group, while in the remainder there was no statistically significant difference. Using the outcome of deterioration in ADL among survivors, day hospital patients showed reduced odds of deterioration compared with those receiving no comprehensive elderly care (0.60; 0.38-0.97; P <0.05). When resource use was examined the day hospital group showed trends towards reductions in hospital bed use and placement of survivors in institutional care. Nine studies comparing treatment costs indicated that day hospital attendance was a more expensive option, although only two analyses took into account long-term care costs. REVIEWER'S CONCLUSIONS: Medical day hospital care for the elderly appears to be more effective than no intervention but may have no clear advantage over other forms of comprehensive elderly medical services. PMID- 10796661 TI - Oral pentoxifylline for treatment of venous leg ulcers. AB - BACKGROUND: Healing of venous leg ulcers is improved by the use of compression bandaging but some venous ulcers do not respond to compression therapy. Pentoxifylline, a drug which helps blood flow, has been used to treat venous leg ulcers but to date there has been no systematic review. OBJECTIVES: To assess the effects of pentoxifylline ('Trental 400') for treating venous leg ulcers, when compared with placebo, or in comparison with other therapies, in the presence or absence of compression therapy. SEARCH STRATEGY: We searched the Cochrane Peripheral Vascular Diseases and Wounds Groups specialised registers (date of search August 1999), and reference lists of relevant articles. We hand searched relevant journals and conference proceedings, and contacted Hoechst (the manufacturer of the drug) and experts in the field. SELECTION CRITERIA: Randomised trials comparing pentoxifylline with placebo or other therapy in the presence or absence of compression, in patients with venous leg ulcers. DATA COLLECTION AND ANALYSIS: Details from eligible trials were extracted and summarised by one reviewer using a coding sheet. Data extraction was independently verified by one other reviewer. MAIN RESULTS: Nine trials involving 572 adults were included. The quality of trials was variable. Eight trials compared pentoxifylline with placebo; in five of these trials patients received compression therapy. In one trial pentoxifylline was compared with defibrotide in patients who also received compression. By pooling eight trials that compared pentoxifylline with placebo (with or without compression) it was found pentoxifylline was more effective than placebo in terms of complete healing or significant improvement (relative risk for healing with pentoxifylline compared with placebo 1.41, 95% confidence interval 1.19 -1.66). Pentoxifylline and compression was more effective than placebo and compression (relative risk for healing with pentoxifylline 1.30, 95% confidence interval 1.10-1.54). Combination of similar trials using compression obtained a number needed to treat (NNT) of 7 (95%confidence interval 4-17). A comparison between pentoxifylline and defibrotide found no difference in healing rates. More adverse effects were reported in the pentoxifylline group, although this was not statistically significant (relative risk for adverse effects with pentoxifylline 1. 25, 95% confidence interval 0.87-1.80). Nearly half of the adverse effects were reported to be gastro-intestinal. REVIEWER'S CONCLUSIONS: Pentoxifylline appears to be an effective adjunct to compression bandaging for treating venous ulcers. There was no cost effectiveness data available and healthcare commissioners may therefore conclude that it not be considered a routine adjunct. Pentoxifylline in the absence of compression may be effective for treating venous ulcers in the absence of compression, although the evidence should be cautiously interpreted. The majority of adverse effects are likely to be tolerated by patients, and gastrointestinal disturbances (indigestion, diarrhoea and nausea) are the most frequent adverse effect. PMID- 10796662 TI - Beds, mattresses and cushions for pressure sore prevention and treatment. AB - OBJECTIVES: To assess the effectiveness of pressure relieving beds, mattresses and cushions (support surfaces) in the prevention and treatment of pressure sores. SEARCH STRATEGY: Searches of 19 databases, hand searching of journals, conference proceedings, and bibliographies. SELECTION CRITERIA: Randomised controlled trials evaluating support surfaces for the prevention or treatment of pressure sores. There was no restriction on articles based on language or publication status. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of study quality was undertaken by two reviewers independently. Trials with similar patients, comparisons, and outcomes were pooled. Where pooling was inappropriate, trials are discussed in a narrative review. MAIN RESULTS: PREVENTION: 29 RCTs of support surfaces for pressure sore prevention were identified. Some high specification foam mattresses were more effective than 'standard' hospital foam mattresses in moderate-high risk patients. Pressure relieving mattresses in the operating theatre reduced the incidence of pressure sores post-operatively. The relative merits of alternating and constant low pressure, and of the different alternating pressure devices are unclear. Seat cushions and simple, constant low-pressure devices have not been adequately evaluated. Limited evidence suggests that low air loss beds reduce the incidence of pressure sores in intensive care. TREATMENT: 6 RCTs of support surfaces for pressure sore treatment were identified. There is good evidence that air fluidised and low air loss beds improve healing rates. Seat cushions have not been adequately evaluated. 2 RCTs evaluated surfaces for both prevention and treatment in the same trial. REVIEWER'S CONCLUSIONS: PREVENTION - There is good evidence of the effectiveness of high specification foam over standard hospital foam, and pressure relief in the operating theatre. Treatment - There is good evidence of the effectiveness of air-fluidised and low air loss devices as treatments. Overall, however, it is impossible to determine the most effective surface for either prevention or treatment. PMID- 10796663 TI - Skin grafting for venous leg ulcers. AB - BACKGROUND: Venous leg ulceration is a common, recurring, disabling condition affecting up to 1% of adults. Treatment is aimed at healing with simple dressings and compression bandages / stockings. Unfortunately in some cases this treatment is unsuccessful with ulcers remaining open for months or years. In order to stimulate healing, some clinicians use skin grafts. These skin grafts may be taken from the patients own uninjured skin (e.g. thigh), may be grown from the patient's skin cells into a dressing, (both known as autografts) or applied as a sheet of bioengineered skin grown from a donor cells (known as an allograft). Preserved skin from other animals, e.g. pigs, have also been used and these are known as xerografts. OBJECTIVES: To assess the effectiveness of skin grafts in the treatment of venous leg ulcers. SEARCH STRATEGY: We searched the Cochrane Wounds Groups specialised register (date of search October 1999), and reference lists of relevant articles. We hand searched relevant journals and conference proceedings, and contacted experts in the field. SELECTION CRITERIA: Randomised controlled trials evaluating skin grafts in the treatment of venous leg ulcers. There was no restriction on articles based on language or publication status. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of study quality was undertaken by two reviewers independently. Trials with similar patients, comparisons, and outcomes were pooled. MAIN RESULTS: Seven RCTs of skin grafts for venous leg ulcers were identified. In 6 trials patients also received compression bandaging. Two trials (98 patients) evaluated split thickness autografts, three trials (92 patients) evaluated cultured keratinocyte allografts, one compared tissue engineered skin (artificial skin) with a dressing (309 patients) and one compared it with a split thickness skin graft (7 patients, 13 ulcers). The trials comparing artificial skin with a dressing reported a significantly higher proportion of ulcers healing with artificial skin. There was insufficient evidence from the remaining trials to determine whether other types of skin grafting increased the healing of venous ulcers. REVIEWER'S CONCLUSIONS: There is limited evidence that artificial skin used in conjunction with compression bandaging, increases the chance of healing a venous ulcer compared to compression alone. Further research is needed to assess whether other forms of skin grafts increase ulcer healing. PMID- 10796664 TI - Corticosteroids for acute severe asthma in hospitalised patients. AB - BACKGROUND: Corticosteroids are currently used routinely in the management of acute severe asthma. The optimal dose and route of administration continues to be debated. Some investigators have reported a greater benefit of higher doses of corticosteroids in the management of severe asthma, while others have not. OBJECTIVES: To determine whether higher doses of systemic corticosteroids (oral, intravenous or intramuscular) are more effective than lower doses in the management of patients with acute severe asthma requiring hospital admission. SEARCH STRATEGY: Randomised controlled trials were identified from the Cochrane Airways Group Asthma Register. In addition, primary authors and content experts were contacted to identify eligible studies. Bibliographies from included studies, known reviews and texts were also searched. SELECTION CRITERIA: Studies were selected for inclusion in the review if they met the following broad inclusion criteria: described as randomised controlled trials, included patients with acute severe asthma, compared different doses of corticosteroids (any route) in 2 or more treatment arms, and had a minimum period of follow up of 24 hours. Two reviewers independently assessed the studies for inclusion and disagreement was resolved by third party adjudication. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers if the authors were unable to verify the validity of information. Missing data were obtained from authors or calculated from other data presented in the paper. The data were analysed as weighted mean differences (WMD) for primary pulmonary function outcomes using a fixed effects model. For the purposes of the review, three broad categories of corticosteroid dose (equivalent dose of methylprednisolone in 24 hours) were defined in advance: low dose (< or = 80 mg), medium dose (> 80 mg and < or = 360 mg) and high dose (> 360 mg). There were thus 3 main comparison groups: low versus medium dose, medium versus high dose and low versus high dose. MAIN RESULTS: Nine trials were included; a total of 344 adult patients have been studied (96 with low dose, 85 with medium dose and 163 with high dose corticosteroids). Only 6 trials provided sufficient data for the meta-analysis. There were no clinically or statistically significant differences detected in % predicted FEV1 among comparison groups after 24, 48 or 72 hours. At 48 hours, the weighted mean difference was -3.3% predicted (95% confidence interval -12.4 to + 5.8) for the low vs medium dose comparison, -1. 9% predicted (95% CI -8.1 to + 4.3) for the medium vs high dose comparison and + 0.5% predicted (95% CI - 7.8 to + 8.8) for the low vs high dose comparison. There appeared to be no significant differences in side effects or rates of respiratory failure among the varying doses of corticosteroids. REVIEWER'S CONCLUSIONS: No differences were identified among the different doses of corticosteroids in acute asthma requiring hospital admission. Low dose corticosteroids (< or = 80 mg/day of methylprednisolone or < or = 400 mg/day of hydrocortisone) appear to be adequate in the initial management of these adult patients. Higher doses do not appear to offer a therapeutic advantage. PMID- 10796665 TI - Speleotherapy for asthma. AB - BACKGROUND: Speleotherapy, the use of subterranean environments, is a therapeutic measure in the treatment of chronic obstructive airways diseases. It is virtually unknown in the UK or the US, but has considerable widespread use in some Central and Eastern European countries. OBJECTIVES: To review evidence for the efficacy of speleotherapy in the treatment of asthma. SEARCH STRATEGY: We searched electronic databases (Medline, Embase, Cochrane Airways group database), contacted speleotherapy centres and experts in the field, hand searched proceedings, and checked bibliographies of articles obtained to identify possible relevant publications. SELECTION CRITERIA: We included controlled clinical trials (i.e., both randomized and those not reporting the method of allocation) that compared clinical effects of speleotherapy with another intervention or no intervention in patients with chronic asthma. DATA COLLECTION AND ANALYSIS: Information concerning patients, interventions, results, and methodology were extracted in standardized manner by two independent reviewers and summarized descriptively. MAIN RESULTS: Three trials including a total of 124 asthmatic children met the inclusion criteria, but only one trial had reasonable methodological quality. Two trials reported that speleotherapy had a beneficial short-term effect on lung function. Other outcomes could not be assessed in a reliable manner. REVIEWER'S CONCLUSIONS: The available evidence does not permit a reliable conclusion as to whether speleo-therapeutic interventions are effective for the treatment of chronic asthma. Randomized controlled trials with long-term follow-up are necessary. PMID- 10796666 TI - Domicilary oxygen for chronic obstructive pulmonary disease. AB - BACKGROUND: Long-term domiciliary oxygen therapy has become one of the major forms of treatment for hypoxaemic chronic obstructive pulmonary disease (COPD) patients. OBJECTIVES: To determine the effect of domiciliary oxygen therapy on survival and quality of life in patients with COPD. SEARCH STRATEGY: Randomised controlled trials (RCTs) were identified using the Cochrane Airways Group COPD register using the search terms: (home OR domiciliary) AND oxygen. SELECTION CRITERIA: Any RCT in patients with hypoxaemia and COPD that compared long term domiciliary or home oxygen therapy with a control treatment. DATA COLLECTION AND ANALYSIS: Data extraction was performed independently by two reviewers. MAIN RESULTS: Four randomised controlled trials were identified. Data from none of these trials could be aggregated because of differences in trial design and patient selection. NOTT 1980, continuous oxygen therapy versus nocturnal oxygen therapy: there was a significant improvement in mortality after 24 months (Peto odds ratio 0.45, 95% confidence interval 0.25 to 0.81). MRC 1981, domiciliary oxygen therapy versus no oxygen therapy: there was a significant improvement over five years in mortality in the group receiving oxygen therapy (Peto odds ratio 0.42, 95% confidence interval 0.18 to 0.98). Fletcher 1992, nocturnal oxygen versus no oxygen in patients with COPD and arterial desaturation at night: there was no difference in mortality at 36 months. Gorecka 1997, long term oxygen versus no oxygen in moderate hypoxaemia: there was no effect on survival for up to three years of follow up. REVIEWER'S CONCLUSIONS: Long term oxygen therapy improved survival in a selected group of COPD patients with severe hypoxaemia (arterial PO2 less than 8.0 kPa). Long term oxygen did not appear to improve survival in patients with moderate hypoxaemia or in those with only arterial desaturation at night. PMID- 10796667 TI - Nicotine for Alzheimer's disease. AB - BACKGROUND: Nicotine is a cholinergic agonist that acts, not only post synaptically, but also releases pre-synaptic acetylcholine, and in animal models has been shown to reverse spatial memory decline in rats with lesion in the medial septal nucleus and to show recovery on memory in aged monkeys. Nicotine also has effects on other transmitters like serotonin (5HT), dopamine, or GABA. On the other hand, because nicotine has serious adverse effects, especially concerning cardiovascular risks in elderly people, and also on sleep and behavior, there are several important reasons to conduct a systematic review to assess the clinical efficacy and safety of nicotine in patients with AD. OBJECTIVES: The aim of this review is to determine whether there is evidence of beneficial effect, and to assess its safety profile, when nicotine is used for Alzheimer's disease. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Issue 2, 99) was searched using the terms nicotin* and alzheimer*. Three references to trials were deemed suitable for inclusion but are awaiting consideration while the investigators are contacted. SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with nicotine patches or administration of nicotine intravenously was administered for more than a day and compared to placebo in people with Alzheimer's disease. DATA COLLECTION AND ANALYSIS: As no trials were suitable for inclusion, no data have been extracted or pooled in a meta-analysis. One trial is awaiting consideration, and if included in an update of this review, any available data will be incorporated. MAIN RESULTS: The poor quality of the trials did not allow any synthesis of results across studies. However, the data available in trials considered are compatible with nicotine producing harm, no change or improvement. REVIEWER'S CONCLUSIONS: This review is not able to provide reliable evidence that nicotine is a useful treatment for Alzheimer's disease. PMID- 10796668 TI - Surgery for small asymptomatic abdominal aortic aneurysms. AB - BACKGROUND: An aneurysm is an abnormal ballooning of an artery. One site in which this occurs is in the abdominal aorta, which is the major artery running through the abdomen. Some abdominal aortic aneurysms (AAA) present as an emergency and require surgery; others remain asymptomatic. Treatment of asymptomatic aneurysms depends on a number of factors, one of which is size. The risk of rupture increases with aneurysm size. Large asymptomatic aneurysms (>6 cm diameter) are operated on; small aneurysms (<4 cm diameter) have regular ultrasound to monitor growth. OBJECTIVES: The objective of this review was to compare the mortality, quality of life and cost effectiveness of early surgical repair with routine ultrasound surveillance in patients with an AAA of between 4-6 cm diameter. SEARCH STRATEGY: Trials were identified through searching the Cochrane Peripheral Vascular Diseases Group trials register and the reference lists of relevant articles. The reviewers also contacted investigators in the field and hand searched recent conference proceedings. SELECTION CRITERIA: Randomised controlled trials in which men and women with asymptomatic AAA of diameter 4-6 cm were randomly allocated to early surgery, or ultrasound surveillance at least once every 12 months. Outcome measures had to include mortality, quality of life or financial costs. DATA COLLECTION AND ANALYSIS: Data were abstracted by one reviewer and checked by others. Due to the small number of trials at present no tests of heterogeneity or sensitivity analyses were performed. MAIN RESULTS: Only one trial, the UK Small Aneurysm Trial, fulfilled the criteria for inclusion. This trial found no differences in mortality between the early surgery and surveillance groups at two, four and six years following randomisation (six years Peto OR 1.01 [95% CI 0.77-1.31]). Mean health service costs were higher in the surgery than the surveillance group, difference 1,064 pounds per patient [95% CI 796-1332]. Quality of life remained similar in the two groups but early surgery patients thought they were healthier and had less pain one year after randomisation. There were not enough patients in the trial to allow analysis of subgroups by, for example, age or aneurysm size. REVIEWER'S CONCLUSIONS: The results from the one trial to date indicate that patients with asymptomatic AAA of 4-5.5 cm should normally have regular ultrasound surveillance accompanied by surgical intervention for aneurysms which grow rapidly (>1 cm per year) or reach 5.5 cm. The results are awaited of a major trial in progress in the USA. PMID- 10796669 TI - Steroids for treating tuberculous pleurisy. AB - BACKGROUND: TB of the pleura is associated with inflammation and fibrosis. Steroids could reduce the effects of the inflammation, but the immunosuppression could make patients vunerable. OBJECTIVES: This review aims to summarise the evidence about the effects of corticosteroids in patients with TB of the pleura, and explores if HIV status is associated with differences in effect estimates. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Library, MEDLINE, and EMBASE. Lists of references from review articles and primary studies were scanned and experts in the field of tuberculosis were contacted. SELECTION CRITERIA: Randomised and quasi-randomised trials evaluating the effects of adjunctive corticosteroids in patients diagnosed with TB pleurisy were sought. Both beneficial and adverse effects were noted. DATA COLLECTION AND ANALYSIS: Two authors independently applied inclusion criteria, assessed trial quality and extracted the relevant data. MAIN RESULTS: Three small trials met the inclusion criteria( total participants n=236), conducted in only HIV negative patients, and with insufficient power to examine death as an outcome. There was no difference in residual lung function between steroid and control groups at completion of treatment. The point estimates for secondary outcomes tended towards benefit with steroids rather than harm, but none were significant; number with pleural fluid (RR 0.28, 95% CI 0.06 to 1.34), number with pleural thickening (RR 0.76, 95% 0.48 to 1.21), and number with pleural adhesions (RR 0.30, 95% CI 0.03 to 2.66). Adverse effects were few and did not result in treatment being discontinued. REVIEWER'S CONCLUSIONS: There is insufficient evidence to know whether steroids are effective in tuberculous pleural effusion. PMID- 10796670 TI - Ziprasidone for schizophrenia and severe mental illness. AB - BACKGROUND: Typical antipsychotic drugs are widely used as the first line treatment for people with schizophrenia. However, the atypical class of antipsychotic drugs are making important inroads into this approach. Atypical is a widely used term used to describe some antipsychotics which have a low propensity to produce movement disorders, sedation and raised serum prolactin. There is some suggestion that the different adverse effect profiles of the atypical antipsychotic group make them more acceptable to people with schizophrenia. Ziprasidone is one of the newer atypicals with a high serotonin/dopamine receptor affinity. OBJECTIVES: To determine the effects of ziprasidone compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), EMBASE (1980-1999), MEDLINE (1966-1999), LILACS (1982-1996), PSYNDEX (1977-1995) and PsycLIT (1974 1999) were undertaken. In addition, pharmaceutical databases on the Dialog Corporation Datastar and Dialog services were searched. References of all identified studies were searched for further trials. Pharmaceutical companies and authors of trials were contacted. SELECTION CRITERIA: All randomised controlled trials that compared ziprasidone to other treatments for schizophrenia and schizophrenia-like psychoses were included by independent assessment. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. Data were excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the risk ratio (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data, weighted mean differences were calculated (WMD). All data were inspected for heterogeneity. MAIN RESULTS: Data for this compound range from very short (1 week) studies of the intramuscular preparation, to trials lasting over six months. For measures of mental state ziprasidone seems more effective than placebo (RR 0.8 CI 0.7-0.9) and as effective as haloperidol (RR 0.8 CI 0.7-1). It is less likely than haloperidol to cause movement disorders (RR 0.4 CI 0.2-0.6), but may cause more nausea and vomiting (RR 2.1 CI 1.2-3.8). The injected form of the drug may cause more pain at the injection site than haloperidol (RR 5.3 CI 1.3-22). REVIEWER'S CONCLUSIONS: Currently data are limited. Ziprasidone may be an effective antipsychotic with less extrapyramidal effects than haloperidol. It also may, however, cause more nausea and vomiting than the typical drugs, and, at present, there is no data suggesting that it is different to other atypical compounds. Well planned, conducted and reported long term randomised trials are needed if ziprasidone is to be accepted into everyday use. PMID- 10796671 TI - Zotepine for schizophrenia. AB - BACKGROUND: Typical antipsychotic drugs are widely used as first line treatment for people with schizophrenia. The atypical class of antipsychotic drugs, however, is making important inroads into this approach and zotepine is one such compound. It is a dopamine antagonist and claimed to be to be particularly effective for negative symptoms OBJECTIVES: To determine the effects of zotepine compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), CINAHL (1982-1999), The Cochrane Library (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), EMBASE (1980-1999), Dialog Corporation Datastar service (1999), MEDLINE (1966-1999), and PsycLIT (1974-1999) were undertaken. References of all identified studies were searched for further trials. Knoll Pharmaceuticals and authors of trials were contacted. SELECTION CRITERIA: All randomised clinical trials that compared zotepine to other treatments for people with schizophrenia or other psychoses were included. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. Data were excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the relative risk (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) and numbers needed to harm (NNH), were calculated on an intention-to-treat basis. For continuous data, weighted mean differences were calculated (WMD). All data were inspected for heterogeneity. MAIN RESULTS: All outcomes were short term (4-12 weeks). Limited data suggest that zotepine is an antipsychotic, at least as effective as typical drugs. Mental state measures of 'no clinically important improvement' favour zotepine when compared to other active drugs (n=356, RR 0.8 CI 0.7-0.9, NNT 7 CI 4-22). About one third of people in both zotepine and control groups left the studies before trial completion. Zotepine may result in less movement disorder adverse effects than typical antipsychotic drugs. Trials have not highlighted clear differences between zotepine and other atypical drugs. REVIEWER'S CONCLUSIONS: Zotepine may be a valuable addition to the increasing ranks of atypical antipsychotic drugs. More data from already existing studies is urgently needed to increase the confidence in the findings of this review. New data from well planned, conducted and reported long term pragmatic randomised trials are necessary. Otherwise clinical use of zotepine will be based on speculation on the meaning of the findings of short explanatory trials for everyday practice. PMID- 10796672 TI - Pimozide for schizophrenia or related psychoses. AB - BACKGROUND: Pimozide was first formulated in the late 1960s and marketed for the care of those with schizophrenia or related psychoses such as delusional disorder. OBJECTIVES: To assess the effects of pimozide for people with schizophrenia, non-affective psychotic mental illness and delusional disorder in terms of clinical, social and economic outcomes. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1995), The Cochrane Schizophrenia Group's Register, EMBASE (1980-1995), Janssen-Cilag UK's register of studies (1999), MEDLINE (1966-1995), PsycLIT (1974-1995), hand-searching the references of all included studies and contacting the manufacturers of the compound. SELECTION CRITERIA: All randomised trials relating to people with schizophrenia, or similar disorders comparing pimozide to other drug treatments were sought. Studies where randomisation was implied rather than stated were included if they did not change the results. Primary outcomes were clinically significant change in global function, mental state, relapse, hospital admission, death, adverse events and acceptability of treatment. DATA COLLECTION AND ANALYSIS: Studies were selected, rated and data extracted. For dichotomous data Relative Risks (RR) based on a random effects model with the 95% confidence intervals (CI) were estimated. The number needed to treat statistic (NNT) was calculated where indicated. Analysis was by intention-to-treat. MAIN RESULTS: This review currently includes 34 studies focusing on those with schizophrenia, none on people with delusional disorder. Few people have been randomised to pimozide versus placebo, but data from three longer term studies does suggest that the active drug prevents relapse (RR 0.59 CI 0.4-0.8, NNT 4 CI 2-13). Pimozide has similar efficacy to that of typical antipsychotic drugs for the outcomes of change in global functioning, mental state, relapse and leaving the study early. People allocated to pimozide did not have a higher mortality than those taking other antipsychotics. Pimozide was more likely to cause parkinsonian tremor (RR 1.6 CI 1.1-2.3, NNH 6 CI 3-44) and lead to a requirement for antiparkinsonian medication more frequently (RR 1.8, CI 1.2-2.6, NNH 3 CI 2-5) than other drugs. It was, however, less likely to cause sedation (RR 0.38 CI 0.2-0.7, NNH 6 CI 4-16). REVIEWER'S CONCLUSIONS: Although there are shortcomings in the data there is enough overall consistency, over different outcomes and time scales, to confirm that pimozide is a drug with similar efficacy to other more commonly used antipsychotics such as chlorpromazine for those with schizophrenia. There are no data to support or refute its use for those with delusional disorder. PMID- 10796673 TI - Nasal decongestants for the common cold. AB - BACKGROUND: The common cold is a major and recurrent cause of morbidity, affecting children and adults two or more times each year. Nasal congestion is its commonest symptom, and many therapies are marketed for its relief. There is no meta-analysis of controlled clinical trials on the effects of nasal decongestants for the common cold. OBJECTIVES: To assess the efficacy of nasal decongestants at reducing the symptom of nasal congestion in adults and children with the common cold and to identify possible adverse effects associated with their use. SEARCH STRATEGY: MEDLINE, EMBASE and Current Contents were searched up to May 1999. Searching the Cochrane Acute Respiratory Infections Group's trials register, following the review citation trail from other references and contacting known principal investigators and pharmaceutical companies was also done. SELECTION CRITERIA: Randomised, placebo controlled trials of single-active oral and topical nasal decongestants in adults and children suffering from the common cold. DATA COLLECTION AND ANALYSIS: Data were independently extracted by two reviewers (DT and LB). All outcomes variables were continuous. Subjective outcomes were normalised to a common scale and a weighted mean difference (WMD) was calculated. A standardised mean difference (SMD) was calculated for the objective outcomes. MAIN RESULTS: Four studies in adults, but none in children, fitted all inclusion criteria. There was a significant 13% decrease in subjective symptoms after decongestant compared with placebo. This was supported by a significant decrease in nasal airways resistance. Repeated doses of nasal decongestant were not significantly better than placebo at relieving symptoms of nasal congestion. REVIEWER'S CONCLUSIONS: A single dose of nasal decongestant in the common cold is moderately effective for the short term relief of congestion in adults, while there is no evidence available to show benefit after repeated use over several days. These medications are not recommended for use in young children with the common cold. PMID- 10796674 TI - Glucocorticoids for croup. AB - BACKGROUND: Since the last meta-analysis in 1989, a number of randomised trials on the benefit of glucocorticoids have been published, resulting in an increasing interest in the use of glucocorticoids to treat outpatients with croup. The objective of this review was to provide evidence to guide clinicians in their treatment of patients with croup, to examine the effectiveness of glucocorticoids in these patients, and to identify areas of uncertainty for future research. OBJECTIVES: To determine the effect of glucocorticoids for children with croup. SEARCH STRATEGY: We searched The Cochrane Controlled Trials Register, MEDLINE (January 1966 to August 1997) and Excerpta Medica/EMBASE (January 1974 to August 1997). We also contacted (by mail) authors of identified croup trials published in the last five years to inquire about other trials, published or unpublished. SELECTION CRITERIA: Meta-analysis of randomised controlled trials that examine the effectiveness of glucocorticoid treatment in children with croup. DATA COLLECTION AND ANALYSIS: Data were extracted using a structured form, which captured patient status (inpatient or outpatient), intervention and control, with the name of the drug, route of administration and dose. Data were also collected on the primary outcome measures comprised of a clinical croup score at baseline (as well as any other subsequent assessment times), length of stay (hours), patients status improved (yes/no), and use of co-interventions. The quality of the trials was assessed using empirically derived items that involved scales and components. Two researchers (TPK, MA) then selected studies as being potentially relevant based on a review of the titles and abstracts, if available. The complete text of these studies was then retrieved. All studies that had been retrieved were reviewed independently by two reviewers (AS, TPK). Data were extracted by one reviewer (MA) and checked for accuracy by a second reviewer (TPK). Two observers independently assessed quality (MA, JK), and inter rater agreement was measured by the intra class correlation. Differences were resolved by consensus. MAIN RESULTS: Twenty-four studies were deemed relevant for inclusion (N=2221). Glucocorticoid treatment was associated with an improvement in the croup severity score at 6 hours with an effect size of -1.0 (95% confidence interval -1.5 to -0.6) and at 12 hours -1.0 (-1.6 to -0. 4); at 24 hours this improvement was no longer significant (-1.0, -2. 0 to 0.1). There was a decrease in the number of adrenaline treatments needed in children treated with glucocorticoids: a decrease of 9% (95% confidence interval 2 to 16%) among those treated with budesonide and of 12% (4 to 20%) among those treated with dexamethasone. There was also a decrease in the length of time spent in accident and emergency (-11 hours, 95% confidence interval -18 to 4 hours), and for inpatients hospital stay was reduced by 16 hours (-31 to 1 hour). Publication bias seems to play a part in these results. REVIEWER'S CONCLUSIONS: Dexamethasone and budesonide are effective in relieving the symptoms of croup as early as 6 hours after treatment. Fewer co-interventions are used and the length of time spent in hospital is decreased in patients treated with glucocorticoids. PMID- 10796675 TI - Homoeopathic Oscillococcinum for preventing and treating influenza and influenza like syndromes. AB - BACKGROUND: Influenza is a highly infectious viral disease that is particularly common in the winter months. Conventional management options are limited to bed rest and treatment of complications such as secondary bacterial infections. Oscillococcinum is a patented, commercially available homoeopathic medicine. The rationale for its use in influenza comes from the homoeopathic principle of 'let like be cured by like'. The medicine is manufactured from wild duck heart and liver, a well-known reservoir for influenza viruses. OBJECTIVES: To determine whether homoeopathic Oscillococcinum or similar medicines are more effective than placebo in the prevention and treatment of influenza and influenza-like syndromes. SEARCH STRATEGY: The registry of randomised trials for the Cochrane Complementary Medicine Field was searched using the term "homeopathy" with "influenza", "respiratory tract", "infection", "cough", "virus" and "fever". The manufacturers of Oscillococcinum were contacted for information about other trials. Date of the most recent search: February 1999. SELECTION CRITERIA: Placebo-controlled trials of Oscillococcinum or homeopath-prepared influenza virus, influenza vaccine or avian liver in the prevention and treatment of influenza and influenza-like syndromes. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data and assessed methodological quality independently. MAIN RESULTS: Seven studies were included in the review, three prevention trials (n=2265) and four treatment trials (n=1194). Only for two studies was there sufficient information to complete data extraction fully. There was no evidence that ic treatment can prevent influenza-like syndrome (relative risk 0.64, 95% confidence interval 0.28 to 1.43). Oscillococcinum treatment reduced length of influenza illness by 0.28 days (95% confidence interval 0.50 to 0.06). Oscillococcinum also increased the chance of a patient considering treatment effective (relative risk 1.08; 95% CI 1.17, 1). REVIEWER'S CONCLUSIONS: Oscillococcinum probably reduces the duration of illness in patients presenting with influenza symptoms. Though promising, the data are not strong enough to make a general recommendation to use Oscillococcinum for first-line treatment of influenza and influenza-like syndrome. Further research is warranted but required sample sizes are large. Current evidence does not support a preventative effect of homeopathy in influenza and influenza-like syndromes. PMID- 10796676 TI - Haemophilus influenzae oral vaccination against acute bronchitis. AB - OBJECTIVES: To assess the effects of an oral whole cell nontypeable Haemophilus influenzae (NTHi) vaccine in protecting against recurrent episodes of bronchitis. SEARCH STRATEGY: We searched the Cochrane trials register, MEDLINE, Extramed, ISI Current Contents, Carl Uncover and contacted investigators of the studies. SELECTION CRITERIA: Randomised trials comparing the effects of an oral monobacterial NTHi vaccine on patients with recurrent exacerbations of bronchitis were included when there was overt matching of the vaccine and placebo groups on clinical grounds. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data and assessed trial quality independently from original records and publications for incidence and severity of bronchitis episodes and carriage rate of nontypeable Haemophilus influenzae in the upper respiratory tract every three months following vaccination. MAIN RESULTS: Six trials were included in the study with a total of 440 participants. Oral vaccination using a monobacterial whole cell killed nontypeable Haemophilus influenzae significantly reduced the incidence of bronchitic episodes at 3 months (Weighted Mean Difference [WMD] 6.694; 95% confidence interval [CI] 6.963 - -6.424; p < 0.01) and 6 months (WMD 4.496; 95% CI 4.664 - -4.327; p < 0.01) following vaccination. The effect diminished by 9 months. The severity of exacerbations in the treatment group was likewise reduced by 58% at 3 months (Peto OR = 0. 42; 95% CI 0.16 1.13), and 65% at 6 months (Peto OR = 0.35; 95% CI 0. 16 0.75) following vaccination. REVIEWER'S CONCLUSIONS: Vaccination, in the autumn, of patients with recurrent exacerbations of bronchitis reduced the number and severity of exacerbations over the winter months. A large clinical trial to assess longer term prognosis needs to be completed. PMID- 10796677 TI - Intravenous naftidrofuryl for critical limb ischaemia. AB - BACKGROUND: Peripheral arterial disease affects 5% of men and women by late middle age. In approximately 25% of those affected the condition will progress to critical limb ischaemia (rest pain, ulceration and gangrene) within five years. The vasoactive drug naftidrofuryl is a serotononergic receptor antagonist which may be beneficial in the treatment of severe lower limb disease such as critical limb ischaemia. OBJECTIVES: To determine whether the vasoactive drug, naftidrofuryl, when administered intravenously is effective in alleviating symptoms and reducing progression of disease in patients with critical limb ischaemia. SEARCH STRATEGY: Randomised controlled trials were identified using the search strategy of the Peripheral Vascular Diseases Review group, including MEDLINE and EMBASE searches and reviewing reference lists in papers. In addition, publications were identified through pharmaceutical companies. SELECTION CRITERIA: All randomised controlled trials of critical limb ischaemia in which patients were randomly allocated to intravenous naftidrofuryl or control (either pharmacological, inert placebo or conservative therapy) were included. Patients with intermittent claudication were not eligible for inclusion. Trials were selected by one reviewer and checked independently. DATA COLLECTION AND ANALYSIS: Fifteen trials were identified, but eight were excluded because of poor methodology. The seven remaining trials involved a total of 229 participants from five different countries. The following outcomes were reported: pain reduction, rest pain/necrosis, progression of disease in terms of incidence of surgical reconstruction/amputation, mortality and side effects. On extraction of the data, odds ratios and weighted mean differences were estimated where appropriate. MAIN RESULTS: Treatment with naftidrofuryl tended to show reduction of pain evaluated by both analogue score and analgesic consumption, but the effect was statistically non-significant (weighted mean difference -0.42, 95% confidence interval -1.19 to 0.35). Similarly, improvement in rest pain or skin necrosis occurred, but these effects were also not significant. The effect on mean ankle systolic ankle pressure was inconclusive. REVIEWER'S CONCLUSIONS: Based on the results of these trials, it cannot be confirmed that intravenous naftidrofuryl is effective in the treatment of patients with critical limb ischaemia. However, these results were based on trials of generally low methodological quality which had only a small number of participants, the duration of treatment was extremely short and the methods varied between the trials. The wide range of endpoints effectively precluded any meaningful pooling of the results. Intravenous naftidrofuryl was withdrawn as a treatment for peripheral arterial disease in 1995 because of reported side effects. PMID- 10796678 TI - Telemedicine versus face to face patient care: effects on professional practice and health care outcomes. AB - BACKGROUND: Telemedicine is the use of telecommunications technology for medical diagnosis and patient care. From its beginnings telemedicine has been used in a variety of health care fields, although widespread interest among healthcare providers has only now become apparent with the development of more sophisticated technology. OBJECTIVES: To assess the effects of telemedicine as an alternative to face-to-face patient care. SEARCH STRATEGY: We searched the Effective Practice and Organisation of Care Group's specialised register, The Cochrane Library, MEDLINE (1966-August 1999), EMBASE (to 1996), Cinahl (to August 1999), Inspec (to August 1996), Healthstar (1983-1996), OCLC, Sigle (to 1999), Assia, SCI (1981 1997), SSCI (1981-1997), DHSS-Data. We hand searched the Journal of Telemedicine and Telecare (1995-1999), Telemedicine Journal (1995-1999) and reference lists of articles. We also hand searched conference proceedings and contacted experts in countries identified as having an interest in telemedicine. SELECTION CRITERIA: Randomised trials, controlled before and after studies and interrupted time series comparing telemedicine with face-to-face patient care. The participants were qualified health professionals and patients receiving care through telemedicine. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Seven trials involving more than 800 people were included. One trial was concerned with telemedicine in the emergency department, one with video-consultations between primary health care and the hospital outpatients department, and the remainder were concerned with the provision of home care or patient self-monitoring of chronic disease. The studies appeared to be well conducted, although patient numbers were small in all but one. Although none of the studies showed any detrimental effects from the interventions, neither did they show unequivocal benefits and the findings did not constitute evidence of the safety of telemedicine. None of the studies included formal economic analysis. All the technological aspects of the interventions appear to have been reliable, and to have been well accepted by patients. REVIEWER'S CONCLUSIONS: Establishing systems for patient care using telecommunications technologies is feasible, but there is little evidence of clinical benefits. The studies provided variable and inconclusive results for other outcomes such as psychological measures, and no analysable data about the cost effectiveness of telemedicine systems. The review demonstrates the need for further research and the fact that it is feasible to carry out randomised trials of telemedicine applications. Policy makers should be cautious about recommending increased use and investment in unevaluated technologies. PMID- 10796679 TI - Nursing record systems: effects on nursing practice and health care outcomes. AB - BACKGROUND: A nursing record system is the record of care planned and/or given to individual patients/clients by qualified nurses or other caregivers under the direction of a qualified nurse. Nursing record systems may be an effective way of influencing nurse practice. OBJECTIVES: To assess the effects of nursing record systems on nursing practice and patient outcomes. SEARCH STRATEGY: We searched The Cochrane Library, MEDLINE, Cinahl, Sigle, and databases of the Royal College of Nursing, King's Fund, the NHS Centre for Reviews and Dissemination, and the Institute of Electrical Engineers up to August 1999; and OCLC First Search, Department of Health database, NHS Register of Computer Applications and the Health Visitors' Association database up to the end of 1995. We hand searched the Journal of Nursing Administration (1971-1999), Computers in Nursing (1984-1999), Information Technology in Nursing (1989-1999) and reference lists of articles. We also hand searched the major health informatics conference proceedings. We contacted experts in the field of nursing informatics, suppliers of nursing computer systems, and relevant Internet groups. SELECTION CRITERIA: Randomised trials, controlled before and after studies and interrupted time series comparing one kind of nursing record system with another, in hospital, community or primary care settings. The participants were qualified nurses, students or health care assistants working under the direction of a qualified nurse and patients receiving care recorded and/or planned using nursing record systems. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Six trials involving 1407 people were included. In three studies of client held records, there were no overall positive or negative effects, although some administrative benefits through fewer missing notes were suggested. A paediatric pain management sheet study showed a positive effect on the children's pain intensity. A computerised nursing care planning study showed a negative effect on documented nursing care planning. A controlled before-and after study of two paper nursing record systems showed improvement in meeting documentation standards. REVIEWER'S CONCLUSIONS: No evidence was found of effects on practice attributable to changes in record systems. Although there is a paucity of studies of sufficient methodological rigour to yield reliable results in this area, it is clear from the literature that it is possible to set up randomised trials or other quasi-experimental designs needed to produce evidence for practice. The research undertaken so far may have suffered both from methodological problems and faulty hypotheses. PMID- 10796680 TI - Antibiotics for treating scrub typhus. AB - BACKGROUND: Scrub typhus is a cause of fever in regions of Asia and the Pacific. Recently, resistance to antibiotics has been reported. OBJECTIVES: To assess treatment regimens for scrub typhus through time to fever resolution and incidence of relapse. SEARCH STRATEGY: Cochrane Controlled Trials Register, MEDLINE, EMBASE, and contacted individual researchers for unpublished data. SELECTION CRITERIA: Randomised and pseudorandomised studies; patients diagnosed with scrub typhus defined by authors; any comparison of antibiotic regimens for treating scrub typhus. DATA COLLECTION AND ANALYSIS: Trial quality was assessed, and data abstracted by both reviewers. MAIN RESULTS: Three trials met the inclusion criteria. One small trial compared tetracycline against chloramphenicol. The other two trials compared doxycycline with tetracycline, and showed little difference in the proportion febrile at 48 hours, with no relapses after either drug. REVIEWER'S CONCLUSIONS: Tetracycline and doxycycline seem to be effective in treating scrub typhus. Further research is required to identify appropriate treatment in areas where Orientia tsutsugamushi resistant to doxycycline has been reported. PMID- 10796681 TI - Antipyretic measures for treating fever in malaria. AB - OBJECTIVES: Fever control measures are commonly used in treating malaria. Some researchers have suggested that fever reduction may prolong malaria illness. We aim to assess whether antipyretic measures in malaria influences outcome, measured by length of illness, parasitaemia, and occurrence of convulsions. SEARCH STRATEGY: We searched the Cochrane infectious Diseases Group Trial Register, the Cochrane Controlled Trial Register, and other electronic bibliographies, and contacted researchers and organizations working in this field. SELECTION CRITERIA: Randomised or pseudo-randomised trials which compared antipyretic drugs with mechanical or no antipyretic measures in patients with slide-confirmed malaria. DATA COLLECTION AND ANALYSIS: Inclusion criteria were independently applied by two reviewers. We extracted data from selected trials using a standard form. Weighted mean difference with 95% confidence interval was calculated for continuous data. MAIN RESULTS: Three randomised trials with pooled 128 adults and children with falciparum malaria; all unblinded; allocation concealment unclear in two. Inconsistent pattern of fever clearance between trials, but malaria cure rate reported to be similar between intervention and control in all trials. Mean parasite clearance time reported to be similar in one trial but longer in paracetamol group in two trials: sample size in one trial was too small to conclude anything (n=7), while the other trial was difficult to evaluate. REVIEWER'S CONCLUSIONS: There is no statistically significant data to draw any conclusions. The clinical significance of preliminary report suggesting that antipyretic drugs prolong malaria parasitaemia was not confirmed. PMID- 10796682 TI - Interventions for preventing reactions to snake antivenom. AB - BACKGROUND: Antivenom is used to neutralise snake bite toxins in people showing evidence of envenomation. It is made from animal sera, and adverse effects, including life threatening anaphylaxis, are common. OBJECTIVES: To assess the effects of drugs given routinely with snake antivenom to prevent adverse effects. SEARCH STRATEGY: Cochrane controlled trials register; contact with researchers in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials testing routine adrenaline (epinephrine), antihistamines, or corticosteroids. DATA COLLECTION AND ANALYSIS: The two authors applied the inclusion criteria, assessed trial quality, and extracted the data. We sought additional data from trialists where required. MAIN RESULTS: One trial in Sri Lanka (n = 105) giving adrenaline with polyspecific antivenom showed fewer adverse reactions in the adrenaline group, and this effect was preserved when stratified for severity. One trial in Brazil (n = 101) using three types of Bothrops antivenom showed no benefit of antihistamine drugs. REVIEWER'S CONCLUSIONS: Routine prophylactic adrenaline for polyvalent antivenom known to have high adverse event rates seems sensible, based on this one trial. If clinicians believe local factors do not justify routine adrenaline, then they should test their belief in a randomised trial. Antihistamine appears to be of no obvious benefit in preventing acute reactions from antivenoms. PMID- 10796683 TI - Inhaled vs oral steroids for adults with chronic asthma. AB - OBJECTIVES: To determine therapeutically equivalent doses of inhaled versus oral steroids for adults with chronic asthma. SEARCH STRATEGY: The Cochrane Airways Group trials register was searched using the terms: (drug delivery systems OR ((nebuli* OR inhal* OR MDI) AND oral*)) AND ( steroid* OR corticosteroid* OR glucocorticoid* OR beclomethasone OR betamethasone OR fluticasone OR cortisone OR dexamethasone OR hydrocortisone OR prednisolone OR prednisone OR triamcinolone). SELECTION CRITERIA: Randomised controlled trials were selected of at least 4 weeks duration and included patients over the age of 15 years with chronic asthma. Trials compared inhaled steroids and oral prednisolone or prednisone; where the maximum dose for inhaled steroids was 2000 mcg/day and prednisolone 60 mg (on alternate days). DATA COLLECTION AND ANALYSIS: Two independent reviewers screened 1285 titles and abstracts from the electronic search, bibliography searches and other contacts. Of these, 10 trials met previously defined inclusion criteria. Two reviewers independently extracted study characteristics, and outcome measures. MAIN RESULTS: All trials were small and no data could be pooled. Carry-over effects were present in at least one cross-over trial. Data from six trials produced the same pattern, in which prednisolone 7.5-12 mg/day appeared to be as effective as inhaled steroid 300-2000 mcg/day. In two trials, inhaled steroid 300-400 mcg/day was more effective than prednisolone 5 mg/day. All doses of inhaled steroid appeared to be more effective than alternate day doses of prednisolone up to 60 mg on alternate days. Side-effect data were reported too variably to permit comparisons. A 30% incidence was reported in one study in patients receiving prednisolone 5 mg/day, none were reported in patients on inhaled steroids. REVIEWER'S CONCLUSIONS: A daily dose of prednisolone 7.5-10 mg/day appears to be equivalent to moderate-high dose inhaled corticosteroids. Side-effects may be present on low doses, so if there is no alternative to oral steroids, the lowest effective dose should be prescribed. PMID- 10796684 TI - Nedocromil sodium versus cromoglycate for the pre-treatment of exercise induced bronchoconstriction in asthma. PMID- 10796685 TI - Early emergency department treatment of acute asthma with systemic corticosteroids. AB - BACKGROUND: The airway edema and secretions associated with acute asthma are most effectively treated with anti-inflammatories such as corticosteroids delivered by inhaled, oral, intravenous or intra-muscular routes. There is an unresolved debate about the use of systemic corticorticoids in the early treatment of acute asthma for emergency department patients. OBJECTIVES: To determine the benefit of treating patients with acute asthma with systemic corticosteroids within an hour of presenting to the emergency department (ED). SEARCH STRATEGY: Randomised controlled trials were identified from the Cochrane Airways Group Asthma Register. Primary authors and content experts were contacted to identify eligible studies. Bibliographies from included studies and known reviews were searched. SELECTION CRITERIA: Only randomised controlled trials (RCTs) or quasi-randomised trials were eligible for inclusion. Studies were included if patients presenting to the ED with acute asthma were treated with IV/IM or oral corticosteroids (CS) vs. placebo within 1 hour of arrival and either admission rate or pulmonary function results were reported. DATA COLLECTION AND ANALYSIS: Trial selection, data extraction and quality assessment were carried out independently by two reviewers, and confirmed with corresponding authors. MAIN RESULTS: Twelve studies involving 863 patients (435 corticosteroids; 428 placebo) were included. Early use of CS for acute asthma in the ED significantly reduced admission rates (N = 11; pooled OR: 0.40, 95% CI: 0.21 to 0.78). This would correspond with a number needed to treat of 8 (95% CI: 5 to 21). This benefit was more pronounced for those not receiving systemic CS prior to ED presentation (N = 7; OR: 0.37, 95% CI: 0.19 to 0.70) and those with more severe asthma (N = 7; OR: 0.35, 95% CI: 0.21 to 0. 59). Oral CS therapy in children was particularly effective (N = 3; OR: 0.24, 95% CI: 0.11 to 0.53); no trials in adults used the oral route. Side effects were not significantly different between corticosteroid treatments and placebo. REVIEWER'S CONCLUSIONS: Use of corticosteroids within 1 hour of presentation to an ED significantly reduces the need for hospital admission in patients with acute asthma. Benefits appear greatest in patients with more severe asthma, and those not currently receiving steroids. Children appear to respond well to oral steroids. PMID- 10796686 TI - Interventions for helping patients to follow prescriptions for medications. AB - BACKGROUND: BACKGROUND: Efforts to assist patients with adherence to prescribed, self-administered medications might improve the benefits and efficiency of health care. OBJECTIVES: OBJECTIVE: To update an ongoing review summarising the results of randomised controlled trials (RCTs) of interventions to help patients follow prescriptions for medications, focusing on trials that measured both adherence and clinical outcomes. SEARCH STRATEGY: Computerised searches to July 1998 in MEDLINE, CINAHL, The Cochrane Library, International Pharmaceutical Abstracts (IPA), PsychInfo, Sociofile, and HSTAR; bibliographies in articles on patient adherence; articles in the reviewers' personal collections; and contact with authors. SELECTION CRITERIA: Articles were selected if they reported an unconfounded RCT of an intervention to improve adherence with prescribed medications, measuring both medication adherence and treatment outcome, with at least 80% follow-up of each group studied and, for long-term treatments, at least six months follow-up for studies with positive initial findings. DATA COLLECTION AND ANALYSIS: Information on study design features, interventions and controls, and findings were extracted by one reviewer (PM) and a research assistant and confirmed by two of the other reviewers. The studies were too disparate to warrant meta-analysis. MAIN RESULTS: For short-term treatments, one study, of counselling and written information, showed an effect on adherence and clinical outcome. Ten of 19 interventions for long-term treatments reported in 17 RCTs were associated with improvements in adherence, but only nine interventions led to improvements in treatment outcomes. Almost all of the interventions that were effective for long-term care were complex, including combinations of more convenient care, information, counselling, reminders, self-monitoring, reinforcement, family therapy, and other forms of additional supervision or attention. Even the most effective interventions did not lead to large improvements in adherence and treatment outcomes. Two studies showed that telling patients about adverse effects of treatment did not affect their adherence. REVIEWER'S CONCLUSIONS: The full benefits of medications cannot be realised at currently achievable levels of adherence. Current methods of improving adherence for chronic health problems are mostly complex and not very effective. More studies of innovative approaches to assist patients to follow medication prescriptions are needed. PMID- 10796687 TI - Routine versus selective antifungal administration for control of fungal infections in patients with cancer. AB - BACKGROUND: Systemic fungal infection is considered to be an important cause of morbidity and mortality in cancer patients, particularly those with neutropenia. Antifungal drugs are often given prophylactically, or to patients with persistent fever. OBJECTIVES: The objective of this review was to assess the effect of antifungal drugs in cancer patients with neutropenia. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (October 1997), MEDLINE (to September 1997) and the reference lists of articles. We searched the proceedings of the ICAAC, General Meeting of the ASM (from 1990 to 1995), and the 7th European Congress of Clinical Microbiology and Infectious Diseases (1995) and contacted researchers in the field. SELECTION CRITERIA: Randomised trials of amphotericin B, AmBisome, fluconazole, ketoconazole, miconazole, or itraconazole compared with placebo or no treatment in cancer patients with neutropenia. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility, methodological quality and abstracted data. MAIN RESULTS: Twenty-five trials involving 2912 patients were included. In 19 trials, prophylactic or later treatment with antifungal drugs had no effect on mortality (odds ratio 0.92, 95% confidence interval 0.75 to 1.14). Only amphotericin showed a significant benefit (odds ratio 0.58, 95% confidence interval 0.37 to 0.93) based on seven trials, but the studies were small and the difference in number of deaths was only 15. Overall it would be necessary to treat 59 patients (95% confidence interval 37 to 131) with an antifungal drug to prevent one case of fungal invasion in surviving patients, although only amphotericin and fluconazole showed a clear beneficial effect. Antifungal treatment decreased fungal colonisation and the need for additional antifungal therapy, but there was heterogeneity across the trials. REVIEWER'S CONCLUSIONS: Routine prophylactic or later therapy with antifungal drugs in cancer patients with neutropenia does not appear to have a beneficial effect on mortality and only a modest effect on fungal invasion. PMID- 10796688 TI - Pharmacotherapy for hypertension in the elderly. AB - OBJECTIVES: To quantify the long-term effects of antihypertensive drug therapy on morbidity and mortality in the elderly. To characterize co morbid risk profiles of trial participants. SEARCH STRATEGY: Electronic search of WHO-ISH Collaboration register (August 1997), The Cochrane Library (1997; Issue 1), MEDLINE (1966 to April 1997) and two Japanese databases (1973-1995); references from reviews, trials and 10 previously published meta-analyses; and experts. SELECTION CRITERIA: Randomized controlled trials of at least one year duration in hypertensive elders (at least 60 years old) assessing antihypertensive drug therapy and providing morbidity and mortality data. DATA COLLECTION AND ANALYSIS: At least two independent reviewers abstracted data on morbidity and mortality results and trial characteristics. The following outcomes were assessed: total mortality; coronary heart disease (CHD) mortality; combined CHD morbidity and mortality; cerebrovascular mortality; combined cerebrovascular morbidity and mortality; cardiovascular mortality; combined cardiovascular morbidity and mortality; and drop outs due to side effects of treatment. MAIN RESULTS: Fifteen trials including 21,908 elderly subjects were identified. The average prevalence of cardiovascular risk factors, cardiovascular disease, and competing co morbid diseases was lower among trial participants than the general population of hypertensive elderly persons. Most subjects were 60 to 80 years old. Most trials were conducted in Western, industrialized countries and evaluated diuretic and beta-blocker therapies. Event rates per 1000 participants over approximately 5 years indicated that antihypertensive drug therapy was beneficial. Cardiovascular morbidity and mortality was reduced from 177 to 126 events (95% CI of the difference 31 to 73). Cardiovascular mortality was reduced from 69 to 50 deaths (95% CI of the difference 9 to 31). Total mortality was reduced from 129 to 111 deaths (95% CI of difference 4 to 28). The data from the three trials restricted to persons with isolated systolic hypertension indicated a significant benefit: cardiovascular morbidity and mortality over approximately 5 years was reduced from 157 to 104 events per 1000 participants (95% CI of the difference 12 to 89). Numbers of participants who dropped out of trials secondary to adverse drug effects were often not reported. The four trials that did report this data showed a wide variation in drop out rates ranging from no significant differences between treatment and control groups to as many as one out of four patients dropping out due to side effects of treatment. REVIEWER'S CONCLUSIONS: Randomized controlled trials establish that treating healthy older persons with hypertension is highly efficacious. Benefits of treatment with low dose diuretics or beta blockers are clear for persons in their 60s to 70s with either diastolic or systolic hypertension. Differential treatment effects based on patient risk factors, pre-existing cardiovascular disease and competing co-morbidities could not be established from the published trial data. PMID- 10796689 TI - Barbiturates for acute traumatic brain injury. AB - BACKGROUND: Raised intracranial pressure (ICP) is an important complication of severe brain injury, and is associated with a high mortality rate. Barbiturates are believed to reduce intracranial pressure by suppressing cerebral metabolism, thus reducing cerebral metabolic demands and cerebral blood volume. However, barbiturates also reduce blood pressure and therefore may adversely effect cerebral perfusion pressure. OBJECTIVES: To assess the effects of barbiturates in reducing raised intracranial pressure, mortality and morbidity in people with acute traumatic brain injury. To quantify any side effects resulting from the use of barbiturates. SEARCH STRATEGY: The review draws largely on the search strategy developed for the Cochrane Injuries Group as a whole. However, in addition the Cochrane Library was searched in December 1996 using the text terms "barbiturate*," "pentobarb*," "phenobarb*," "head," and "brain." An updated search was done in April 1999. SELECTION CRITERIA: Randomised or quasi randomised trials of any one or more of the barbiturate class of drugs (amobarbital, barbital, hexobarbital, mephobarbital, methohexital, murexide, pentobarbital, phenobarbital, secobarbital, thiobarbiturate) where study participants had a clinically diagnosed acute traumatic brain injury of any severity. DATA COLLECTION AND ANALYSIS: The reviewer extracted the data and assessed the quality of allocation concealment in the trials. MAIN RESULTS: The pooled relative risk for death (barbiturate vs no barbiturate) was 1.09 (95%CI 0.81 to 1.47). The pooled effect of barbiturates on adverse neurological outcome, measured using the Glasgow Outcome Scale (death, persistent vegetative state or severe disability) was 1.15 (95% 0.81 to 1.64). Two studies examined the effect of barbiturate therapy on intracranial pressure. In the study by Eisenberger et al, a smaller proportion of patients in the barbiturate group had uncontrolled ICP (68% vs 83%). The relative risk for uncontrolled ICP was 0.81 (95%CI 0.62 to 1.06). Similarly, in the study by Ward et al, mean ICP was lower in the barbiturate treated group. Barbiturate therapy results in an increase in the occurrence of hypotension (RR=1.80 95%CI 1.19 to 2.70). For every four patients treated one will develop clinically significant hypotension. Mean body temperature was significantly lower in the barbiturate treated group. Schwartz et al compared pentobarbital with mannitol in the control of intracranial pressure. Pentobarbital was less effective than mannitol for control of raised ICP. 68% of patients in the pentobarbital treated group required a second drug for the treatment of raised intracranial pressure compared to 39% in the mannitol treated group (RR=1.75 95%CI 1.05 to 2.92). There was no substantial difference in mortality between the two study groups (RR=1.18 95%CI 0.73 to 1.92). REVIEWER'S CONCLUSIONS: There is no evidence that barbiturate therapy in patients with acute severe head injury improves outcome. Barbiturate therapy results in a fall in blood pressure in 1 in 4 treated patients. The hypotensive effect of barbiturate therapy will offset any ICP lowering effect on cerebral perfusion pressure. PMID- 10796690 TI - Ovulation induction with urinary follicle stimulating hormone versus human menopausal gonadotropin for clomiphene-resistant polycystic ovary syndrome. AB - BACKGROUND: The risks of multiple pregnancy and ovarian hyperstimulation syndrome (OHSS) are increased in women with clomiphene resistance WHO group 2 dysfunction undergoing ovulation induction as well as the risk of spontaneous abortion if conception takes place. Semi-purified preparations of FSH have been developed in an effort to reduce the impact of exogenous LH, relatively high levels of which are present in human menopausal gonadotropin (hMG). Ovulation induction in women with clomiphene resistant WHO group 2 dysfunction who often have clinical features of polycystic ovarian syndrome (PCOS), is a major challenge. The risks of multiple pregnancy and ovarian hyperstimulation syndrome (OHSS) are increased in this population. There also appears to be an increased risk of spontaneous abortion in those who conceive, perhaps associated with elevated LH levels. Semi purified preparations of FSH have been developed in an effort to reduce the impact of exogenous LH, relatively high levels of which are present in human menopausal gonadotrophins. OBJECTIVES: To determine the effectiveness of daily FSH versus daily hMG in women with clomiphene-resistant polycystic ovary syndrome (PCOS), in terms of rates of pregnancy and moderate to severe ovarian hyperstimulation syndrome (OHSS). SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: All RCTs relevant to the clinical question were selected. DATA COLLECTION AND ANALYSIS: A diverse search strategy was employed, including hand search of 43 core journals from 1966 to the present, bibliographies of relevant trials, MEDLINE database, abstracts from North American and European meetings and contact with authors of relevant papers. Relevant data were extracted independently by two reviewers using the standardized data extraction sheet. Validity was assessed in terms of method of randomisation, completeness of follow up, presence or absence of crossover and co-intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were generated using the Peto modified Mantel-Haenszel technique. Statistical heterogeneity was assessed using x2. MAIN RESULTS: No significant benefit was demonstrated from semi-purified FSH versus hMG in terms of pregnancy rate: common odds ratio per patient 0.66 (95% CI 0.35-1.24) and per cycle 0.89 (95% CI 0.51-1.53). FSH appeared to be associated with a reduction in moderate to severe OHSS: common odds ratios 0.2 (95% CI 0.09-0.46). REVIEWER'S CONCLUSIONS: In women with PCOS, no significant difference could be demonstrated between FSH and hMG, in terms of pregnancy rate. However, given similar cost, potential advantages in terms of purity and a possible reduction in OHSS risk, highly purified or recombinant FSH are likely to be widely adopted in the future. Further research should consider live birth as a primary clinical outcome, given concerns over the association between high androgen and LH levels with spontaneous abortion risk. PMID- 10796691 TI - Oil-soluble versus water-soluble media for assessing tubal patency with hysterosalpingography or laparoscopy in subfertile women. AB - BACKGROUND: There has been debate in the literature for more than 40 years as to whether flushing of the Fallopian tubes enhances fertility and whether this presumed therapeutic effect is greater with oil-soluble media than with water soluble media. A meta-analysis of the therapeutic role of oil-soluble contrast media at hysterosalpingography was published in March 1994 (Watson 1994). This Cochrane Review is an expansion and update of that overview. OBJECTIVES: To evaluate the effect of flushing the Fallopian tubes with oil- or water-soluble contrast media on subsequent pregnancy rates in infertility patients. SEARCH STRATEGY: 1. For randomised controlled trials (RCTs): we searched the Cochrane Subfertility Review Group specialised register of controlled trials. 2. For non randomised controlled studies: Medline and manual search of eight relevant journals. SELECTION CRITERIA: Six RCTs and six non-randomised controlled studies. In two studies tubal flushing was compared with no treatment-controls. Eleven studies, comprising a total of 2635 subjects, compared oil- with water-soluble media. DATA COLLECTION AND ANALYSIS: Independently by first 2 authors for: 1. Methodological trial characteristics 2. Characteristics of participants 3. OUTCOMES: the main studied outcome was pregnancy rate per patient MAIN RESULTS: In comparison with no treatment, a significant benefit on pregnancy rates was seen by flushing the tubes with oil-soluble media (OR 1.80, 95% CI 1.29-2.50) but not with water-soluble media (OR 0.87, 95% CI 0.50-1.52). In the direct comparison of oil- and water-soluble media the former lead to significantly higher pregnancy rates (OR 1.92, 95% CI 1.60-2.29). The treatment effect appeared similar in RCTs and non-RCTs. Subgroup analysis suggested the therapeutic benefit was greatest for patients suffering from unexplained infertility and least for those with tubal factor infertility. REVIEWER'S CONCLUSIONS: Flushing of the tubes with oil-soluble media increases subsequent pregnancy rates in infertility patients. The hypothesis that tubal "plugs" are a cause of proximal tubal occlusion and that oil-soluble media may flush them out, is supported by new techniques such as falloposcopy. Clinicians should consider flushing the tubes with OSCM before contemplating more invasive therapies. PMID- 10796692 TI - Growth hormone for in vitro fertilization. AB - BACKGROUND: Ovulation induction protocols for in vitro fertilization (IVF) are constantly under review and revision in an attempt to decrease gonadotropin requirements while improving follicular recruitment and pregnancy rate. Most studies of the effect of Growth Hormone on ovulation induction have investigated normally ovulating infertile women. Attention has begun to turn to its effect on women who fail to produce an optimal number of follicles in response to ovulation induction. OBJECTIVES: To assess the effectiveness of growth hormone adjuvant therapy for women undergoing ovulation induction prior to IVF in two patient groups: a) those with no previous history of poor response and b) those with a history of poor response. SEARCH STRATEGY: This review has drawn on the search strategy developed for the Subfertility Group as a whole. Relevant trials were identified in the Group's Specialised Register of Controlled Trials. See Review Group details for more information. SELECTION CRITERIA: All RCTs were included if they addressed the research question posed and provided outcome data for intervention and control subjects. Outcomes of interest included pregnancy, oocyte/embryo number and peak estradiol level. DATA COLLECTION AND ANALYSIS: DATA EXTRACTION: A diverse search strategy was employed, including hand-search of 43 core journals from 1966 to the present, bibliographies of relevant trials, MEDLINE database, abstracts from North American and European meetings and contact with authors of relevant papers. Relevant data were extracted independently by two reviewers using the standardized data extraction sheet. Validity was assessed in terms of method of randomization, completeness of follow-up, presence or absence of crossover and co-intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were generated using the Peto modified Mantel-Haenszel technique. Statistical heterogeneity was assessed using x2. MAIN RESULTS: In women with no previous history of poor stimulation, the common odds ratio for pregnancy per cycle was 0.97 (95% CI 0.34-2. 76) Total dose of gonadotropin and number of oocytes retrieved were similar between treated and placebo groups. In previous poor responders, the common odds ratio for pregnancy per cycle instituted was 2.55 (95% CI 0.64-10.12). Again, no statistically significant difference was noted in gonadotropin dosage or oocyte number obtained, when all studies were considered. REVIEWER'S CONCLUSIONS: Data from 6 small trials suggest that in women with no previous poor response, GH augmentation does not improve the rate of pregnancy. In poor responders, a trend towards improved outcome with GH treatment deserves further study. PMID- 10796693 TI - Vaccines for preventing hepatitis B in health-care workers. AB - BACKGROUND: Hepatitis B causes acute and chronic liver disease and may be prevented by vaccination. OBJECTIVES: To assess the effectiveness and safety of plasma-derived vaccines against acute and chronic hepatitis B in health-care workers in protecting them from hepatitis B infection and its consequences. SEARCH STRATEGY: MEDLINE and Excerpta Medica Database (EMBASE) search using standard Cochrane strategy, Cochrane Library, full text searching of the journal "Vaccine", bibliography of retrieved studies and correspondence with authors, researchers and manufacturers. SELECTION CRITERIA: All original prospective randomised comparisons of yeast-derived vaccines and plasma-derived vaccines against no intervention, placebo, or vaccines against other disease (control vaccines). Assessment of trial quality was made according to: 1. generation of allocation schedule 2. measure(s) taken to conceal treatment allocation 3. drop out of allocated health-care worker participants from the analysis of trial results 4. measures taken to implement double blinding Trial reports were blinded by removal of authors and their affiliation, journal reference, introduction, results, and discussion. DATA COLLECTION AND ANALYSIS: To assess efficacy the incidence rates of acute hepatitis B were observed in the surveillance of the vaccinated and control groups of the trials included in the review. Safety was assessed from side-effect rates, classified as systemic (malaise, nausea, fever, arthralgias, rash, headache) or local (induration and soreness at the site of the inoculation). MAIN RESULTS: Four trials fulfilling the criteria were identified and the data synthesised. All trials compared plasma-derived vaccines versus placebo. Differences in the settings (and level of incidence) between three of the trial settings and Dienstag's led us to stratify our comparison grouping the three trials performed in dialysis units together. After our stratification, the Desmyter, Smuzness and Crosnier group appears to be homogeneous (Chi-square = 0.11, degrees of freedom = 2). Our estimates of effectiveness and safety in the high risk group favour treatment, the OR for cases of HB being 0.34; 95% CI (0.21, 0.55). The analysis also revealed a non-significant trend towards benefit in the lower risk health-care workers (Dienstag trial, OR 0.26 (0.05, 1.30). Overall the evidence strongly favours vaccination (OR=0.33; 95% CI (0.21, 0.53)). There was no difference in the incidence and severity of side-effects between the two arms of the trials. We calculated that it was necessary to vaccinate between 145 (assuming a baseline rate of 10 cases/1000/year) and 7 (for a baseline rate of 200/1000/year) health-care workers with plasma-derived vaccines to avoid one case of hepatitis B. Completeness of trial reporting was not good with all four trials failing to report titre results on antibodies against hepatitis B surface antigen and hepatitis B core antigen in the placebo arms (correspondence with two of the four authors failed to shed light on the reasons for such an omission). All four trials achieved low scores in the four quality dimensions assessed (generation of allocation schedule, measure(s) taken to conceal treatment allocation, exclusion of allocated participants from the analysis of the trial and measures taken to implement and protect double blinding). Mean length of follow-up was 14.5 months. REVIEWER'S CONCLUSIONS: Plasma-derived vaccines appear to be efficacious and safe for use in high risk health-care workers, such as staff of renal dialysis and transplant units. There is some uncertainty concerning the effectiveness of the vaccine in lower risk health-care workers, although the trend is towards benefit. We found no evidence of a long-term protective effect due to the short follow-up time of the four trials included in this review. We found relatively poor standard of trial reporting, possibly related to the age of the trials. PMID- 10796694 TI - Home-based social support for socially disadvantaged mothers. AB - BACKGROUND: Epidemiologic studies indicate that babies born to socio-economically disadvantaged mothers are at higher risk of injury, abuse and neglect, health problems in infancy, and are less likely to have regular well-child care. Home visitation programs have long been advocated as a strategy for improving the health of disadvantaged children. Over the past two decades, a number of randomised trials have examined the effect of home visitation programs on a range of maternal and child health outcomes. The studies in this review evaluate programs which offer additional home based support for socially disadvantaged mothers and their children. OBJECTIVES: Babies born in socio-economic disadvantage are likely to be at higher risk of injury, abuse and neglect, and to have health problems in infancy. The objective of this review was to assess the effects of programs offering additional home-based support for women who have recently given birth and who are socially disadvantaged. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. Date of last search: 26 October 1998. SELECTION CRITERIA: Randomised and quasi-randomised trials of one or more post natal home visits with the aim of providing additional home based support for socially disadvantaged women who had recently given birth, compared to usual care. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Study authors were contacted for additional information. MAIN RESULTS: Eleven studies, involving 2992 families, were included. Most of the trials had important methodological limitations. Seven trial reports are awaiting further assessment. There was a trend towards reduced child injury rates with additional support, although this was not statistically significant (odds ratio 0.74, 95% confidence interval 0.54 to 1.03). There appeared to be no difference for child abuse and neglect (odds ratio 1.12, 95% confidence interval 0.80 to 1.57), although differential surveillance between visited and non-visited families is an important methodological consideration. Babies in the additional support groups were more likely to have complete well-child immunizations. Based on the results of two trials, there was a trend towards reduced hospitalization, although this was not statistically significant. REVIEWER'S CONCLUSIONS: Postnatal home-based support programs appear to have no risks and may have benefits for socially disadvantaged mothers and their children, possibly including reduced rates of child injury. Differential surveillance does not allow easy interpretation of the child abuse and neglect findings. PMID- 10796695 TI - Kinin-enhancing drugs for unexplained subfertility in men. AB - BACKGROUND: Oligo-astheno-teratospermia (sperm of low concentration, reduced motility and increased abnormal morphology) of unknown cause is common and the need for treatment is felt by patients and doctors alike. As a result, a variety of empirical, non-specific treatments have been used in an attempt to improve semen characteristics and fertility. One suggested treatment for idiopathic oligo and/or asthenospermia is the administration of kallikrein (kallidinogenase), a kinin-releasing enzyme (or kininogenase). The kinin biological system is complex and involves kininogen (the substrate), kininogenases (the activating enzymes), kinins (the effectors) and kininases (the inactivating enzymes). All four components of the kinin system have been found in the genitalia and in semen. Kallikrein releases 2 major kinins, kallidin and bradykinin, from seminal plasma kininogens. Activated kinins in semen affect sperm motility and metabolism. In vitro addition of kallikrein to semen has been shown to have a positive effect on sperm motility, sperm velocity, cervical mucus penetration, penetration of zona free hamster eggs and post-thaw survival and motility rate after semen cryopreservation. The latter observation, however, was not confirmed in a more recent comparison of motility stimulants for cryopreserved semen using computerised sperm motion analysis. In vitro treatment of semen with kallikrein has been employed in a clinical context during sperm preparation prior to insemination. Although the kinin system may also be involved in the regulation of spermatogenesis in vivo, a clear mechanism of action is missing. Multiple suggestions on how an increase in kinin levels in the genital tract influences spermatogenesis at the testicular levels have been made by various authors. OBJECTIVES: To determine whether treatment of the male with drugs enhancing kinin levles increases pregnancy rates among couples where failure to conceive has been attributed to idiopathic oligo- and/or asthenospermia. Effects on sperm parameters and sex hormones were studied as secondary outcomes. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched". SELECTION CRITERIA: SIxteen RCTs on the therapeutic use of androgens (clomiphene citrate or tamoxifen) in subfertile men were identified. Six trials were excluded. DATA COLLECTION AND ANALYSIS: Methodological characteristics of trials Baseline characteristics of the studied groups Outcomes: Pregnancy rates, semen parameters (sperm concentration, motility and morphology), endocrinology (serum FSH, testosterone and oestradiol) PMID- 10796696 TI - Oral contraceptive pills for heavy menstrual bleeding. AB - BACKGROUND: Menorrhagia (heavy menstrual bleeding) is a benign yet debilitating social and health condition. The widely accepted clinical definition of menorrhagia is blood loss of 80ml or more per period. This figure is derived from population studies that have shown that the average blood loss is between 30 and 40ml, and 90% of women have blood losses of less than 80ml. Excessive menstrual bleeding is the commonest cause of iron deficiency in the United Kingdom affecting 20-25% of the fertile female population. Menorrhagia is a common problem accounting for 12% of all gynaecological referral in the UK. Ranges of medical therapies are prescribed in order to reduce excessive menstrual blood loss, including prostaglandin synthetase inhibitors, antifibrinolytics, the oral contraceptive pill and other hormones. The combined oral contraceptive pill (OCP) is claimed to have a variety of beneficial, inducing a regular shedding of a thinner endometrium and inhibiting ovulation thus having the effect of treating menorrhagia and providing contraception. OBJECTIVES: To determine whether: 1. the OCP is an effective medical therapy to reduce menorrhagia in both the short term and long term. 2. the effectiveness of combined oral contraceptive pills (OCP) compared with other medical therapies for the treatment of menorrhagia. 3. OCP is a more cost effective method than any other medical treatments of menorrhagia. 4. OCP has fewer side effects than other drugs used for menorrhagia. SEARCH STRATEGY: All publications which describe randomised trials of OCP for the treatment of menorrhagia were obtained using the search strategy developed by the Menstrual Disorders Group. SELECTION CRITERIA: All randomised controlled comparisons of OCP versus other medical therapies, placebo or no treatment for the treatment of menorrhagia. Women of reproductive years with regular heavy periods, measured either objectively or subjectively and greater than, or equal to, two months follow up. DATA COLLECTION AND ANALYSIS: All assessments of the quality of trials and data extraction were performed unblinded by at least two reviewers. Only one trial met the inclusion criteria and none were excluded. The included trial involved a total of 45 women. MAIN RESULTS: As the trial used a cross-over design, only data from the first treatment period (cycles 3 and 4 ) were analysed. The results from all the three mefanamic acid groups were combined. There was no significant difference in menstrual blood loss (MBL) between those patients treated with the OCP and danazol, mefenamic acid or naproxen. REVIEWER'S CONCLUSIONS: The one small study identified [Fraser 1991] found no significant difference between groups treated with OCP, mefenamic acid, low dose danazol or naproxen. Overall, the evidence from the one study identified [Fraser 1991] is not sufficient to adequately assess the effectiveness of OCP. This review was unable to achieve its stated objectives because of the paucity of the data. PMID- 10796697 TI - Ovulation suppression for endometriosis. AB - BACKGROUND: Although the etiology of endometriosis is unknown, several theories exist, the most popular of which is retrograde menstruation. As endometriosis can only be diagnosed by laparoscopy, neither the incidence (annual occurrence) nor the prevalence (proportion of the population affected) of endometriosis is known. The association between endometriosis and infertility isn't clear in Stage I (minimal) and Stage II (mild) endometriosis. Endometriosis appears to be an estrogen dependent condition. At the time of menopause, most endometriosis becomes quiescent. This hormonal dependency prompted researchers to seek agents which would suppress ovarian activity. OBJECTIVES: To determine effectiveness of a) ovulation suppression with danazol, medroxy progesterone acetate, gestrinone, combined oral contraceptive pills and GnRH analogues versus placebo or no treatment and b) any of the above agents versus danazol, for the treatment of endometriosis explained infertility in terms of clinical pregnancy rate. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Four RCTs with five treatment arms compared an ovulation suppression agent with placebo or no treatment. Eight trials were identified comparing a suppressive agent with danazol. DATA COLLECTION AND ANALYSIS: DATA EXTRACTION: A diverse search strategy was employed, including hand-search of 43 core journals from 1966 to the present, bibliographies of relevant trials, MEDLINE database, abstracts from North American and European meetings and contact with authors of relevant papers. Relevant data were extracted independently by two reviewers using the standardised data extraction sheet. Validity was assessed in terms of method of randomisation, completeness of follow-up, presence or absence of crossover and co intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were generated using the Peto modified Mantel-Haenszel technique. Statistical heterogeneity was assessed using x2. MAIN RESULTS: The common odds ratio for pregnancy following ovulation suppression versus placebo or no treatment was 0.83 (95% CI 0.5-1.39). These data were statistically homogeneous although clinical heterogeneity was present. Their consistency in showing no treatment benefit suggests that this group of interventions is ineffective. Common odds ratio for pregnancy following all agents versus danazol was 1.20 (95% CI 0. 85-1.68). Again these data were homogeneous and suggest no significant treatment benefit in terms of pregnancy rate. REVIEWER'S CONCLUSIONS: Given the significant period of amenorrhea associated with ovulation suppression, the lack of treatment benefit demonstrated and the adverse effects commonly associated with these treatments, ovulation suppression cannot be recommended as a standard therapy for endometriosis-associated infertility. PMID- 10796698 TI - Anti-epileptic drugs for preventing seizures following acute traumatic brain injury. AB - BACKGROUND: Seizure activity in the early post-traumatic period following head injury may cause secondary brain damage as a result of increased metabolic demands, raised intracranial pressure and excess neurotransmitter release. OBJECTIVES: To determine the effects of prophylactic anti-epileptic agents for acute traumatic head injury. SEARCH STRATEGY: We searched the Cochrane Injuries Group trials register, Medline and the databases of the Cochrane Stroke Group and Cochrane Epilepsy Group. We also contacted pharmaceutical companies who manufacture anti-epileptic agents, the National Institute of Neurological Disorders and Stroke, Epilepsy Division, and the National Institute of Health, United States. SELECTION CRITERIA: All randomised trials of anti-epileptic agents, in which study participants had a clinically defined acute traumatic head injury of any severity. Trials in which the intervention was started more than eight weeks after injury were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed the quality of the trials. Relative risks and 95% confidence intervals were calculated for each trial on an intention to treat basis, which included pre-drug loading exclusions. As long as statistical heterogeneity did not exist, for dichotomous data, summary relative risks and 95% confidence intervals were calculated using a fixed effects model. Where the source of heterogeneity could obviously be related to allocation concealment, drug type, or drug dose, we stratified the analyses on that dimension. MAIN RESULTS: We identified 10 eligible randomised controlled trials, including 2036 participants. Data are currently unavailable for four unpublished trials, representing 631 participants. For the remaining trials, the pooled relative risk (RR) for early seizure prevention was 0.34 (95% confidence interval 0.21 to 0.54); based on this estimate, for every 100 patients treated, 10 would be kept seizure free in the first week. Seizure control in the acute phase was not accompanied by a reduction in mortality (RR=1.15; 95% confidence interval 0.89 to 1. 51), a reduction in death and neurological disability (RR = 1.49; 95% confidence interval 1.06 to 2.08 for carbamazepine and RR= 0.96; 95% confidence interval 0.72 to 1.26 for phenytoin) or a reduction in late seizures (pooled RR =1.28; 95% confidence interval 0.90 to 1. 81). The pooled relative risk for skin rashes was 1.57 (95% confidence interval 0.57 to 39.88). REVIEWER'S CONCLUSIONS: Prophylactic anti-epileptics are effective in reducing early seizures, but there is no evidence that treatment with prophylactic anti-epileptics reduces the occurrence of late seizures, or has any effect on death and neurological disability. Insufficient evidence is available to establish the net benefit of prophylactic treatment at any time after injury. PMID- 10796699 TI - Somatostatin or octreotide for acute bleeding oesophageal varices. AB - BACKGROUND: Somatostatin and its derivative, octreotide, are often used for emergency treatment of bleeding oesophageal varices in patients with cirrhosis of the liver. The placebo controlled trials have shown varying results, however, and their power has been quite low. An updated systematic review of a previously published meta-analysis was therefore performed. OBJECTIVES: To study whether somatostatin or octreotide improve survival or reduce the number of blood transfusions in patients with suspected or verified acute or recently bleeding oesophageal varices. SEARCH STRATEGY: MEDLINE and The Cochrane Library are searched every three months. Reference lists of articles and authors. SELECTION CRITERIA: All randomised trials comparing somatostatin or octreotide with placebo or no treatment in patients suspected of acute bleeding from oesophageal varices. DATA COLLECTION AND ANALYSIS: The effect variables extracted were: mortality, number of blood transfusions, number with balloon tamponade, number with initial haemostasis and number with rebleeding. Intention to treat analyses were conducted; a random effects analysis was preferred if there was significant heterogeneity between the trials (p<0.10). MAIN RESULTS: The meta-analysis comprised 820 patients. The active drugs had no effect on survival; a total of 91 patients died in the experimental groups vs 85 patients in the control groups (odds ratio 1.04, 95% confidence interval (CI) 0.74 to 1.46). The number of transfusions was less with drugs, the difference between experimental and control therapy was 1.2 units of blood products saved per patient (95% CI 0. 8 to 1.6). There were no significant differences in use of balloon tamponade (odds ratio 0.59, 95% CI 0.21 to 1.70), number of patients failing initial haemostasis (odds ratio 0.66, 95% CI 0.32 to 1.37) or number with rebleeding (odds ratio 0.73, 95% CI 0.30 to 1.79). It should be noted, however, that the trials were heterogeneous with respect to these secondary variables. REVIEWER'S CONCLUSIONS: The effect corresponded to one unit of blood saved per patient. This effect is small and treatment with these drugs in acute bleeding oesophageal varices is therefore of doubtful value. This does not suggest a need for further studies. On the other hand, the confidence interval for the effect on mortality was wide. Hence, a large placebo controlled trial is needed if one wishes to rule out the possibility that a worthwhile effect on mortality may have been overlooked. PMID- 10796700 TI - Spironolactone versus placebo or in combination with steroids for hirsutism and/or acne. AB - BACKGROUND: Hirsutism is the presence of excessive hair growth in women and is an important cosmetic condition often resulting in severe distress. Hirsutism is most often caused by increased production of male sex hormones also known as androgens. It is also affected by increased sensitivity to androgens in the hair follicles, and the secretory glands around the hair follicles, called sebaceous glands. Spironolactone is an antiandrogen and aldosterone antagonist used to treat hirsutism. Since 1978, many studies have been conducted to determine its effectiveness. OBJECTIVES: The objective of this review was to investigate the effectiveness of spironolactone and/or in combination with steroids (oral contraceptive pill included) in reducing excess hair growth and/or acne in women. SEARCH STRATEGY: All publications of randomised controlled trials of spironolactone versus placebo and/or in combination with steroids (oral contraceptive pill included) were identified. Search strategy was developed by the Menstrual Disorder Group. All accessable electronic databases were searched. In addition, all reference lists of relevant trials were searched and drug companies contacted for details of unpublished trials. SELECTION CRITERIA: All randomised controlled comparisons of spironolactone versus: placebo steroids (oral contraceptive pill included)) spironolactone of varying dosages or spironolactone and steroids versus steroids alone when used to reduce hair growth and acne in women. DATA COLLECTION AND ANALYSIS: Four trials were included in the review, seven trials were excluded. Two other trials are awaiting assessment. All included trials were small (no more than 31 participants) randomised and controlled. Only one trial studied acne as an outcome, the remaining three were concerned with hirsutism. One trial investigated spironolactone versus placebo; one trial was a dosage studies of spironolactone; one trial compared spironolactone with spironolactone in combination with dexamethasone; the remaining trial used topical spironolactone for the treatment of acne. Major outcome measures include the following: - subjective observations - Ferriman and Gallwey hair scores - hormonal and biochemical parameters - side effects - sebum production measurement MAIN RESULTS: All sample populations were small and confidence intervals were wide. There was no significant difference between treatments for any of the observed outcomes. REVIEWER'S CONCLUSIONS: The effectiveness of treatment for either acne vulgaris or hirsutism cannot be determined due to the small sample populations involved in the trials. Its value in clinical practice is difficult to assess from currently available research. PMID- 10796701 TI - Corticosteroids for acute traumatic brain injury. AB - BACKGROUND: Traumatic brain injury is a leading cause of death and disability. Corticosteroids have been widely used in treating people with traumatic brain injury. OBJECTIVES: To quantify the effectiveness and safety of corticosteroids in the treatment of acute traumatic brain injury. SEARCH STRATEGY: Electronic sources: MEDLINE, EMBASE, Cochrane Library and specialised database searches. Additional hand searching and contact with trialists. Date of the most recent search June 1999. SELECTION CRITERIA: All randomized controlled trials of corticosteroid use in acute traumatic brain injury with adequate or unclear allocation concealment. DATA COLLECTION AND ANALYSIS: Quality of allocation concealment was scored. Data on numbers of participants randomized, numbers lost to follow up, length of follow up, case fatality rates, disablement, infections and gastrointestinal bleeds were extracted independently and checked. MAIN RESULTS: We identified 19 trials with 2295 randomized participants. The effect of corticosteroids on the risk of death was reported in 16 included trials. The pooled relative risk for the 16 trials was 0.96 (95% confidence interval 0.85 to 1. 08). Pooled risk difference was 1.3% (5.2% to 2.5% more). For the nine trials that reported death or severe disability, the pooled relative risk was 1.01 (0.91 to 1.11). For infections the pooled relative risk was 0.94 (0.76 to 1.16) and for the nine trials reporting gastrointestinal bleeding 1.11 (0.54 to 2.26). Using only those trials with the best quality of allocation concealment, the pooled relative risk estimate for death became closer to unity. REVIEWER'S CONCLUSIONS: Neither moderate benefits nor moderate harmful effects of steroids can be excluded. The widely practicable nature of the drugs and the importance of the health problem suggest that large simple trials are feasible, and worthwhile, to establish whether there are any benefits from corticosteroids in this situation. PMID- 10796702 TI - Techniques for pelvic surgery in subfertility. AB - BACKGROUND: Since the advent of in-vitro fertilisation (IVF) the role of infertility surgery has diminished. This type of surgery is still however widely performed.and there are many different surgical techniques that can be used to repair blocked or damaged Fallopian tubes. Most evidence in this area comes from uncontrolled series. OBJECTIVES: To evaluate surgical techniques for the treatment of tubal infertility (adhesiolysis, salpingostomy, surgery for proximal tubal occlusion, reversal of sterilisation). SEARCH STRATEGY: This review has drawn on the search strategy developed for the Subfertility Group as a whole. Relevant trials were identified from the Group's Register of Controlled Trials. SELECTION CRITERIA: All randomised controlled trials investigating an aspect of infertility surgery technique. Non-randomised data were included for the following topics 1) The role of infertility surgery versus no treatment 2) The role of infertility surgery versus alternative treatments, 3) the role of magnification, 4) The role of the CO2 laser at infertility surgery, 5) the role of operative laparoscopy to perform infertility surgery. Non- RCT data were excluded if :- i) The treatment and control groups differed significantly ii) The treatment and control groups were operated by a different team or in a different institute iii) Where pregnancy outcome data were not given. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the first 2 authors. Differences of opinion were registered and resolved by consensus with the senior author (RL). Two by two tables were generated for each trial for the dichotomous outcome of pregnancy and the effects on pregnancy rate of each study is expressed as an odds ratio with 95% confidence intervals. MAIN RESULTS: 1) INFERTILITY SURGERY VERSUS NO TREATMENT One non RCT compared open adhesiolysis versus no treatment and found significantly more pregnancies in the treatment group compared with the control group. 2) INFERTILITY SURGERY VERSUS ALTERNATIVE TREATMENTS. No RCTs investigated the role of infertility surgery compared with In vitro fertilisation (IVF). There were no studies comparing tubal surgery for proximal tubal occlusion versus hysteroscopic or radiologically controlled recannulation. 3) MAGNIFICATION FOR INFERTILITY SURGERY There was a non significant reduction in pregnancy rate when the operating microscope (magnification x4-x16) was used rather than Loupes (magnification x2-x4.5) in the only RCT to study this. One RCT randomised patients to microsurgery versus a macrosurgical technique involving a prosthesis. There were more pregnancies in the microsurgery group, but this was not significant, and the trial consisted of only 18 participants. All the non-RCT studies comparing microsurgery with macrosurgery had a historical control group. Meta-analysis of studies investigating the role of magnification for adhesiolysis and for salpingostomy revealed a statistically significant increase in pregnancy rates and reduction in ectopic pregnancy rates for microsurgery versus macrosurgery for both procedures. For reversal of sterilisation there was a significant improvement in term pregnancy rates, and a non significant reduction in ectopic rates There was no significant difference between microsurgical and macrosurgical treatment of proximal tubal occlusion for any outcome. 4) THE USE OF LASER AT INFERTILITY SURGERY Two RCTs investigated the use of the CO2 laser at infertility surgery. There was no significant difference in pregnancy outcome after adhesiolysis, or salpingostomy. Two non randomised studies also investigated the role of the laser. Overall there was no significant difference using the CO2 laser compared with standard techniques for adhesiolysis, salpingostomy or reversal of sterilisation. 5) LAPAROSCOPIC INFERTILITY SURGERY Four studies investigated the use of laparoscopic techniques for infertility surgery. There were no RCTs. One study compared laparoscopic versus open PMID- 10796703 TI - Ursodeoxycholic acid for cystic fibrosis-related liver disease. AB - BACKGROUND: Cystic fibrosis-related liver disease peaks in adolescence with up to 20% of patients developing chronic liver disease. Early changes in the liver may ultimately result in end stage liver disease with patients needing transplantation. One therapeutic option currently used is ursodeoxycholic acid. OBJECTIVES: To assess the effectiveness of ursodeoxycholic acid in cystic fibrosis liver disease. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group trials register, and contacted drug companies. Date of the most recent search of the Group's specialised register: June 1999. SELECTION CRITERIA: Randomised and quasi-randomised trials of use of ursodeoxycholic acid for at least three months compared with placebo or no additional treatment in people with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality. MAIN RESULTS: Three trials involving 118 patients were included. The complex study design used in two trials meant that data could only be analysed for subsets of patients. There was no difference in weight change (weighted mean difference -0.496, 95% confidence interval -1.545 to +0.553) based on 30 patients from two trials. Improvement in biliary excretion was reported in only one study and no significant change after treatment was shown. Long term outcomes such as death or need for liver transplantation were not reported. REVIEWER'S CONCLUSIONS: There are few trials assessing the effectiveness of ursodeoxycholic acid. There is insufficient evidence to justify is its routine use in cystic fibrosis. PMID- 10796704 TI - Pergolide for levodopa-induced complications in Parkinson's disease. AB - OBJECTIVES: To compare the efficacy and safety of adjunct pergolide therapy versus placebo in patients with Parkinson's disease suffering from the complications of levodopa therapy. SEARCH STRATEGY: Electronic searches of MEDLINE, EMBASE and the Cochrane Controlled Trials Register. Handsearching of the neurology literature as part of the Cochrane Movement Disorders Group's strategy. Examination of the reference lists of identified studies and other reviews. Contact with Eli Lilly and Company Limited. SELECTION CRITERIA: Randomised controlled trials of pergolide versus placebo in patients with a clinical diagnosis of idiopathic Parkinson's disease and long-term complications of levodopa therapy. DATA COLLECTION AND ANALYSIS: Data was abstracted independently by each author and differences settled by discussion. The outcome measures used included Parkinson's disease rating scales, levodopa dosage, 'off' time measurements and the frequency of drop outs and adverse events. MAIN RESULTS: A large number of small RCTs were identified, but these were part of a large multicentre trial which was eventually published in full. The final publication was used as the only subject for this review. The time patients spent 'off' was reduced by 1.8 hours with pergolide compared with 0.2 hours with placebo (p < 0.001). Dyskinesia developed or deteriorated in 62% of pergolide-treated compared with 25% placebo-treated patients (p < 0. 05). The excess in dyskinesia prevalence and severity resolved by the end of the study with levodopa reduction. Levodopa dose was reduced more in those receiving pergolide (235 mg v 51 mg; p < 0. 001). Pergolide produced significant improvement in Hoehn and Yahr stage (p < 0.05) and both the motor and activities of daily living parts of a modified Columbia rating scale (both p < 0.001). Significantly more patients suffered nausea (24% v 13%; p < 0.001) and hallucinations (14% v 3%; p < 0.01) on pergolide. No difference was found in the numbers remaining on treatment at the end of the study (pergolide 84% v placebo 82%) but withdrawals due to adverse events were greater in those taking pergolide (10% v 4%). REVIEWER'S CONCLUSIONS: Based on this single large multicentre study, pergolide reduces 'off' time and improves impairment and disability due to Parkinson's disease whilst allowing a reduction in levodopa dose. This is at the expense of dopaminergic adverse events. Further trials are required to compare pergolide with the newer dopamine agonists. PMID- 10796705 TI - Pergolide versus bromocriptine for levodopa-induced motor complications in Parkinson's disease. AB - OBJECTIVES: To compare the efficacy and safety of adjunct pergolide therapy versus bromocriptine in patients with Parkinson's disease, already established on levodopa and suffering the long-term complications of therapy. SEARCH STRATEGY: Electronic searches of MEDLINE, EMBASE and the Cochrane Controlled Trials Register. Handsearching of the neurology literature as part of the Cochrane Movement Disorders Group's strategy. Examination of the reference lists of identified studies and other reviews. Contact with Eli Lilly Company and Sandoz Limited. SELECTION CRITERIA: Randomised controlled trials of pergolide versus bromocriptine in patients with a clinical diagnosis of idiopathic Parkinson's disease and long-term complications of levodopa therapy. DATA COLLECTION AND ANALYSIS: Data was abstracted independently by each author and differences settled by discussion. MAIN RESULTS: Three short-term trials fulfilled the inclusion criteria for the review. Pergolide was superior to bromocriptine regarding UPDRS and NYPDS motor and NYPDS ADL scores in two trials. More patients recorded a 'marked' or 'moderate improvement' in clinician's global impression score with pergolide than bromocriptine in two studies. Insufficient evidence on fluctuations and dyskinesia was available to draw any conclusions. No significant differences between the agonists were seen in levodopa dose reduction, drop outs or adverse events. REVIEWER'S CONCLUSIONS: Although pergolide is superior to bromocriptine in reducing motor impairments and disability, no firm conclusions regarding levodopa-induced motor complications can be reached. Levodopa dose reduction, adverse events and withdrawals from treatment are similar for the two agonists. The small advantage of pergolide in efficacy does not take into account its additional cost compared with bromocriptine. PMID- 10796706 TI - Managements for people with disorders of sexual preference and for convicted sexual offenders. AB - BACKGROUND: The reviewers recognise that it may be thought that convicted sex offenders and those with disorders of sexual preference are quite different groups. In combining them within this review we have taken the view that legal process alone should not define the population. Illegal behaviours in one jurisdiction may not be considered so in others. Studies of those who are convicted of sexual offending describe reconviction rates for sexual offences of up to 40-60%. It would seem important to know if there are interventions that might reduce this high rate of re-offending. This review examines antilibidinal management of those who have been convicted of sexual offences or who have disorders of sexual preference. OBJECTIVES: To determine the effectiveness of a range of management techniques to assist people who have disorders of sexual preference and those who have been convicted of sexual offences. SEARCH STRATEGY: Biological Abstracts, the Cochrane Schizophrenia Group Register of Trials, The Cochrane Library, EMBASE, MEDLINE, and PsychLIT were searched. Further references were sought from published trials and their authors. Relevant pharmaceutical manufacturers were contacted. SELECTION CRITERIA: All relevant randomised controlled trials. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed on an intention-to-treat basis. Data were extracted for short and medium term outcomes. MAIN RESULTS: A single trial (McConaghy 1988) found the effect of antilibidinal medication (medroxyprogesterone acetate) plus imaginal desensitisation was no better than imaginal desensitisation for problematic/anomalous sexual behaviour and desire. A relapse prevention programme was trialed by Marques (Marques 1994) and participants were followed up for an average of 3 years. What data there are suggest that although there is no discernable effect on the outcome of sex offending (OR 0.76 CI 0. 26-2.28) those treated with response prevention do have less non-sexual violent offences (OR 0.3, CI 0.1-0.89, NNT 10 CI 5-85). In addition those committing both sexual and violent offences also declined in the response prevention group (OR 0.14 CI 0.02 0.98, NNT 20 CI 10-437). A large pragmatic trial investigated the value of group therapy for sex offenders (Romero 1983). This study finds no effect on recidivism at ten years. REVIEWER'S CONCLUSIONS: It is disappointing to find that this area lacks a strong evidence base, particularly in light of the controversial nature of the treatment and the high levels of interest in the area. The relapse prevention programme did seem to have some effect on violent reoffending but large, well-conducted randomised trials of long duration are essential if the effectiveness or otherwise of these treatments are to be established. PMID- 10796707 TI - Antioxidant vitamin and mineral supplementation for preventing age-related macular degeneration. AB - BACKGROUND: Some observational studies have suggested that people who eat a diet rich in antioxidant vitamins (carotenoids, vitamins C and E) or minerals (selenium and zinc) may be less likely to develop age-related macular degeneration. OBJECTIVES: The aim of this review is to examine the evidence as to whether or not taking vitamin or mineral supplements prevents the development of age-related macular degeneration. SEARCH STRATEGY: We searched the Cochrane Eyes and Vision Group specialised register, the Cochrane Controlled Trials Register - Central, MEDLINE, reference lists of identified reports and the Science Citation Index. We contacted investigators and experts in the field for details of unpublished studies. The most recent searches were conducted in June 1999. SELECTION CRITERIA: All randomised trials comparing an antioxidant vitamin and/or mineral supplement (alone or in combination) to control were included. We included only studies where supplementation had been given for at least one year. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. Currently there is only one published trial included in the review so no data synthesis was conducted. MAIN RESULTS: One trial is included in the review. This was a primary prevention trial in Finnish male smokers with four treatment groups: alpha-tocopherol alone, beta-carotene alone, alpha-tocopherol and beta-carotene, placebo. The add-on maculopathy study was conducted in a subset of the main trial cohort. 269 cases of maculopathy (14 late stage age-related macular degeneration) were identified. There was no association of age-related macular degeneration with treatment. REVIEWER'S CONCLUSIONS: There is no evidence to date that people without age-related macular degeneration should take antioxidant vitamin and mineral supplements to prevent or delay the onset of the disease. The results of five large ongoing trials are awaited. PMID- 10796708 TI - Antioxidant vitamin and mineral supplements for age-related macular degeneration. AB - BACKGROUND: It has been proposed that antioxidants may prevent cellular damage in the retina by reacting with free radicals produced in the process of light absorption. OBJECTIVES: The objective of this review is to assess the effects of antioxidant vitamin and/or mineral supplementation on the progression of age related macular degeneration. SEARCH STRATEGY: The Cochrane Eyes and Vision Group specialised registered, the Cochrane Controlled Trials Register - Central, MEDLINE, the Science Citation Index, and the reference lists of relevant articles were searched. Investigators of included studies were contacted. The most recent searches were performed in December 1999. SELECTION CRITERIA: All randomised trials comparing an antioxidant vitamin and/or mineral supplement (alone or in combination) to control in people with age-related macular degeneration were included. DATA COLLECTION AND ANALYSIS: The reviewer extracted data and assessed trial quality. Due to the small number of trials identified, and variable methods of collecting and presenting outcome data, no statistical summary measure was calculated. MAIN RESULTS: Four published, two unpublished and two ongoing trials were identified. Published trials to date have been small and results inconsistent. Adverse effects and quality of life for people with age-related macular degeneration have not been addressed. REVIEWER'S CONCLUSIONS: The question as to whether people with age-related macular degeneration should take antioxidant vitamin or mineral supplements to prevent progression of the disease has not been answered by research to date. The results of ongoing trials are awaited. PMID- 10796709 TI - Intra-uterine versus cervical insemination of donor sperm for subfertility. AB - BACKGROUND: Artificial insemination with sperm is used to improve the chances of conception for various causes of infertility. Traditionally, sperm is deposited in or around the endocervical canal (cervical insemination - CI). Some studies reported higher pregnancy rates if sperm was deposited in the uterine cavity itself (intrauterine insemination - IUI), but most were uncontrolled. However the cost and the risks (infection and anaphylaxis) of IUI may also be higher. OBJECTIVES: The objective of this review was to assess the effects of depositing donor sperm in the uterine cavity (intrauterine insemination) compared to cervical insemination. SEARCH STRATEGY: The Cochrane Subfertility Review Group specialised register of controlled trials was searched. SELECTION CRITERIA: Randomised trials comparing intrauterine insemination and cervical insemination, using fresh or cryopreserved semen, with or without ovarian hyperstimulation. DATA COLLECTION AND ANALYSIS: Trial quality assessment and data extraction were done independently by two reviewers. MAIN RESULTS: Twelve studies were included. They comprised 697 patients undergoing 2215 treatment cycles. Ten trials used frozen semen, with three using ovarian hyperstimulation. Overall the methodological quality of the trials was low. The overall pregnancy rate per cycle in the intrauterine insemination group was 18% compared to 5% for cervical insemination. When cryopreserved donor sperm was used, the overall chance of pregnancy in spontaneous or clomiphene-corrected cycles was significantly higher with intrauterine insemination. This was irrespective of whether pregnancy rates were calculated on a per cycle (odds ratio 2. 63, 95% confidence interval 1.85 to 3.73) or per patient (odds ratio 3.86, 95% confidence interval 1.81 to 8.25) basis. The greatest benefit appeared in trials with poor pregnancy rates (less than 6%) for cervical insemination. There was no difference in pregnancy rate between intrauterine and cervical insemination when fresh donor sperm was used (odds ratio 0.90, 95% confidence interval 0.36 to 2. 24). REVIEWER'S CONCLUSIONS: Intrauterine insemination appears to be beneficial when cervical insemination using cryopreserved donor sperm has had low pregnancy rates. This applies to spontaneous, clomiphene corrected and gonadotrophin stimulated cycles. However it may offer little benefit where high pregnancy rates have been achieved with cervical insemination. There appears to be no additional benefit from intrauterine insemination when fresh sperm is used for donor insemination. PMID- 10796710 TI - Interventions for tubal ectopic pregnancy. AB - BACKGROUND: The diagnosis of ectopic pregnancy can now often be made by non invasive methods due to sensitive pregnancy tests (in urine and serum) and high resolution transvaginal sonography, which have been integrated in diagnostic algorithms. These algorithms, in combination with the increased awareness and knowledge of risk factors among both clinicians and patients, have enabled an early and accurate diagnosis of ectopic pregnancy. As a consequence, the clinical presentation of ectopic pregnancy has changed from a life threatening disease to a more benign condition. This in turn has resulted in major changes in the options available for therapeutic management. Many treatment options are now available to the clinician in the treatment of tubal pregnancy: surgical treatment, which can be performed radically or conservatively, either laparoscopically or by an open surgical procedure; medical treatment, with a variety of drugs, that can be administered systemically and/or locally by different routes (transvaginally under sonographic guidance or under laparoscopic guidance); expectant management. The choice of a treatment modality should be based on short-term outcome measures (primary treatment success and reinterventions for clinical symptoms or persistent trophoblast) and on long-term outcome measures (tubal patency and future fertility). OBJECTIVES: In the treatment of tubal pregnancy various types of treatments are available: surgical treatment, medical treatment and expectant management. In this review the effects of various treatments are summarized in terms of treatment success, need for reinterventions, tubal patency and future fertility. SEARCH STRATEGY: The Cochrane Menstrual Disorders and Subfertility Group trials register and MEDLINE were searched. SELECTION CRITERIA: Randomized controlled trials comparing treatments in women with ectopic pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extracted independently by two reviewers. Differences were resolved by discussion with all reviewers. MAIN RESULTS: Laparoscopic conservative surgery is significantly less successful than the open surgical approach in the elimination of tubal pregnancy due to a higher persistent trophoblast rate of laparoscopic surgery. Long term follow-up shows similar tubal patency rates, whereas the number of subsequent intrauterine pregnancies is comparable, and the number of repeat ectopic pregnancies lower, although these differences are not statistically significant. The laparoscopic approach is less costly as a result of significantly less blood loss and analgesic requirement, and a shorter duration of operation time, hospital stay, and convalescence time. Compared to laparoscopic conservative surgery (salpingostomy) local methotrexate is not a treatment option. Injection of this drug, both under laparoscopic guidance and under ultrasound guidance, is significantly less successful in the elimination of tubal pregnancy. Systemic methotrexate in a single dose intramuscular regimen is not effective enough in eliminating the tubal pregnancy compared to laparoscopic salpingostomy. This as a result of inadequately declining serum hCG concentrations after one single dose of methotrexate necessitating additional methotrexate injections or surgical interventions. If methotrexate primarily given in a multiple dose intramuscular regimen is compared with laparoscopic salpingostomy no large differences are found in medical outcomes, both short term and long term. However, this treatment regimen is associated with a greater impairment of health related quality of life and is more expensive, due to surgical interventions for clinical signs of tubal rupture, generating additional direct costs due to prolonged hospital stay. Furthermore, indirect costs due to productivity loss are higher. Only in patients with low initial serum hCG concentrations systemic methotrexate leads to costs savings compared to laparoscopic salpingostomy. PMID- 10796711 TI - Timed intercourse versus intra-uterine insemination with or without ovarian hyperstimulation for subfertility in men. AB - BACKGROUND: Although intra-uterine insemination (IUI) is widely used, however its effectiveness remains a matter of debate. Although IUI is less invasive and expensive than IVF or GIFT, it should only be applied if the probability of conception is improved significantly as compared to the natural chance of conceiving. To increase the number of available oocytes at the site of fertilization, controlled ovarian hyperstimulation (COH) can be applied in conjunction with IUI. Uncontrolled studies suggest a beneficial effect of COH in combination with IUI, also when a male factor is present. To be able to draw firm conclusions whether IUI and/or COH improve the probability of conception, several comparisons should be performed in randomized controlled trials (RCTs). OBJECTIVES: To determine for male subfertility whether intrauterine insemination (IUI) improves the probability of conception compared with timed intercourse and whether the addition of controlled ovarian hyperstimulation influences the results. SEARCH STRATEGY: 1. The specialist database of the Cochrane Menstrual Disorders and Subfertility Group. 2. Medline search. 3. Embase search. 4. DDFU search. 5. BIOSIS search. 6. SCIsearch. 7. Manual searching of references mentioned in the obtained studies. 8. Personal communication and write letters to experts (14) in the field. 9. Abstracts of The American Society for Reproductive Medicine and European Society for Human Reproduction and Embryology Meetings. When important information is lacking from the original publications the authors will be contacted. SELECTION CRITERIA: Randomized controlled trials only. DATA COLLECTION AND ANALYSIS: Independently by the first 2 authors: 1. Trial design characteristics. 2. Baseline characteristics of participants. 3. Types of intervention. 4. Outcomes where pregnancy is the outcome of main interest. Number of multiple pregnancies and number of cycles with ovarian hyperstimulation syndrome (OHSS) are secondary outcomes. Analysis of agreement between the two observers was determined for the following items: inclusion or exclusion of a trial, method of randomization, definition of male subfertility, design of the trial, number of pregnancies and completed cycles. Sensitivity analysis is performed. MAIN RESULTS: Seventeen trials fulfilled the selection criteria for this review and were included. Four trials are pending. Crude agreement concerning inclusion or exclusion of trials occurred for 41 of 43 (95%) trials reviewed (kappa 0.90). The included trials comprised 3,662 completed cycles. In natural cycles intrauterine insemination (IUI) significantly improved the probability of conception compared with timed intercourse (TI) (combined odds ratio with 95% confidence intervals: 2.43, 1.54 - 3.83). In cycles with controlled ovarian hyperstimulation (COH) IUI significantly improved the probability of conception also compared with TI (combined odds ratio with 95% confidence intervals: 2.14, 1.30 - 3.51). Despite clinical heterogeneity, these results are based on strong evidence. Intrauterine insemination in cycles with COH improved the probability of conception compared with IUI in natural cycles but significance was not reached (combined odds ratio with 95% confidence intervals: 1.79, 0.98 - 3.25). Comparing IUI in COH-cycles with TI in natural cycles the first treatment modality significantly improved the probability of conception (combined odds ratio with 95% confidence intervals: 6.23, 2.35 - 16.52). REVIEWER'S CONCLUSIONS: Intra-uterine insemination offers couples with male subfertility benefit over timed intercourse, both in natural cycles and in cycles with COH. In the case of a severe semen defect (with more than 1 million motile sperm after semen preparation and no triple sperm defect) IUI in natural cycles should be the treatment of first choice. The value of COH need to be further investigated in RCTs. Mild ovarian hyperstimulation with gonadotrophins is advised in cases with less sever PMID- 10796712 TI - Antipsychotic medication for challenging behaviour in people with learning disability. AB - BACKGROUND: The term 'challenging behaviour', in the absence of psychiatric disorder, encompasses a wide range of behaviours that may be harmful to people or property, may be difficult to manage and may limit access to community facilities. Antipsychotic medications have been used to modify such behaviours in people with learning disability, but there is little evidence to suggest that the benefits outweigh the risks. OBJECTIVES: To determine the effectiveness of antipsychotic medication for people with learning disability and challenging behaviour. SEARCH STRATEGY: Biological abstracts, the Cochrane Library, the Cochrane Schizophrenia Group's Register, EMBASE, MEDLINE, PsycLIT were searched. Further references were sought from published trials and pharmaceutical companies. Trials were reliably identified and data extracted. SELECTION CRITERIA: All randomised controlled trials of antipsychotic medication versus placebo. DATA COLLECTION AND ANALYSIS: Reviewers independently evaluated and analysed data on an intention to treat basis. Data were evaluated at 4 and 8 weeks as longer follow-up data were not available. Reviewers assumed that those subjects lost to follow-up had a bad outcome. MAIN RESULTS: Only three randomised controlled trials could be included in the analyses. These provided no evidence of whether antipsychotic medication helps or harms adults with learning disability and challenging behaviour. REVIEWER'S CONCLUSIONS: There is limited data on this important issue and more research is urgently needed. PMID- 10796713 TI - Interventions for treating chronic pelvic pain in women. AB - BACKGROUND: Chronic pelvic pain is common in women in the reproductive age group and it causes disability and distress and results in significant costs to health services. The pathogenesis of chronic pelvic pain is poorly understood. Often, investigation by laparoscopy reveals no obvious cause for pain. There are several possible explanations for chronic pelvic pain including undetected irritable bowel syndrome, the vascular hypothesis where pain is thought to arise from dilated pelvic veins in which blood flow is markedly reduced and altered spinal cord and brain processing of stimuli in women with chronic pelvic pain. As the pathophysiology of chronic pelvic pain is not well understood, its treatment is often unsatisfactory and limited to symptom relief. Currently, the main approaches to treatment include counselling or psychotherapy, attempting to provide reassurance using laparoscopy to exclude serious pathology, progestogen therapy such as medroxyprogesterone acetate, and surgery to interrupt nerve pathways. OBJECTIVES: We aimed to identify and review treatments for chronic pelvic pain in women in the reproductive years. The review included studies of patients with a diagnosis of pelvic congestion syndrome but excluded those with pain known to be caused by i) endometriosis, ii) primary dysmenorrhoea (period pain), iii) pain due to chronic pelvic inflammatory disease, or iv) irritable bowel syndrome. SEARCH STRATEGY: The search strategy adopted by the Cochrane Menstrual Disorders and Subfertility Group was used. SELECTION CRITERIA: Randomised controlled trials (RCTs) with women who had chronic pelvic pain, excluding endometriosis, primary dysmenorrhoea, pain due to chronic pelvic inflammatory disease, or irritable bowel syndrome. The reviewers were prepared to consider studies of any intervention including lifestyle, physical, medical, surgical and psychological treatments. Outcome measures were pain rating scales, quality of life measures, economic analyses and adverse events. DATA COLLECTION AND ANALYSIS: For each included trial, information was collected regarding the method of randomisation, allocation concealment, blinding, whether an intention to treat analysis could possibly be performed and relevant interventions and outcomes (see previous sections). Data were extracted independently by the two reviewers, using forms designed according to the Cochrane guidelines. MAIN RESULTS: Seven studies were identified of which four were of good methodological quality. One study was reported in a brief abstract only and was excluded. Progestagen (Medroxyprogesterone acetate) was associated with a reduction of pain during treatment. Counselling supported by ultrasound scanning was associated with reduced pain and improvement in mood. A multidisciplinary approach was beneficial for some outcome measures. Adhesiolysis was not associated with an improved outcome apart from where adhesions were severe. REVIEWER'S CONCLUSIONS: Further studies to confirm these observations are needed, together with full reporting of those studies which have been undertaken. Given the prevalence and health care costs associated with chronic pelvic pain in women, randomised controlled trials of other medical, surgical and psychological interventions are urgently required. PMID- 10796714 TI - Nonsteroidal anti-inflammatory drugs for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding is an important cause of ill health in premenopausal women. Although surgery is often used as a treatment, a range of medical therapies are also available. Nonsteroidal anti-inflammatory drugs or prostaglandin synthetase inhibitors reduce prostaglandin levels which are elevated in women with excessive menstrual bleeding and also may have a beneficial effect on dysmenorrhoea. OBJECTIVES: The primary objective of this review was to investigate the effectiveness of non-steroidal anti-inflammatory drugs (NSAIDs) in achieving a reduction in menstrual blood loss in women of reproductive years with heavy menstrual bleeding (HMB). SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Register of Trials, MEDLINE, EMBASE, PsychLIT, Current Contents, Biological Abstracts, Social Sciences Index and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles and drug companies were approached for unpublished data. In most cases, the first author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of individual NSAIDs with either each other, placebo or other medical treatments in women of reproductive years with regular heavy periods measured either objectively or subjectively and with no pathological or iatrogenic (treatment induced) causes for their heavy menstrual blood loss. DATA COLLECTION AND ANALYSIS: Sixteen RCTs were identified that fulfilled the inclusion criteria for this review. The reviewers extracted the data independently and odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data of nine trials. The remaining seven trials were of crossover design with data unsuitable for pooling and their individual results were described in text form. MAIN RESULTS: As a group, NSAIDs were more effective than placebo at reducing heavy menstrual bleeding but less effective than either tranexamic acid or danazol. Treatment with danazol caused a shorter duration of menstruation and more adverse events than NSAIDs but this did not appear to affect the acceptability of treatment. There was a non significant trend towards greater efficacy of NSAIDs compared to oral progestogen (luteal phase) and ethamsylate but no differences were demonstrated between NSAIDs and the progesterone releasing intra-uterine system (IUS) and the oral contraceptive pill, although these results were based on very small studies. There was no evidence of a difference between the individual NSAIDs (naproxen and mefenamic acid) in reducing HMB. REVIEWER'S CONCLUSIONS: NSAIDs reduce heavy menstrual bleeding when compared with placebo but are less effective than either tranexamic acid or danazol. However, adverse events are more severe with danazol therapy. In the limited number of small scale studies suitable for evaluation, no significant difference in efficacy was demonstrated between NSAIDs and other medical treatments such as oral progestogen given in the luteal phase, ethamsylate, oral contraceptive pill and the progesterone releasing IUS. PMID- 10796715 TI - Hormone replacement therapy in postmenopausal women: endometrial hyperplasia and irregular bleeding. AB - BACKGROUND: The decline in circulating oestrogen around the time of the menopause often induces unacceptable symptoms that affect the health and well being of women. Hormone replacement therapy (both unopposed oestrogen and oestrogen and progestogen combinations) is an effective treatment for these symptoms. In women with an intact uterus, unopposed oestrogen may induce endometrial stimulation and increase the risk of endometrial hyperplasia and carcinoma. The addition of progestogen reduces this risk but may cause unacceptable symptoms, bleeding and spotting which can affect adherence to therapy. OBJECTIVES: The objective of this review is to assess which hormone replacement therapy regimens provide effective protection against the development of endometrial hyperplasia and/or carcinoma with a low rate of abnormal vaginal bleeding. SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Register of Trials, MEDLINE, EMBASE, PsychLIT, Current Contents, Biological Abstracts, Social Sciences Index and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles and drug companies were contacted for unpublished data. In most cases, the corresponding author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of unopposed oestrogen therapy, combined continuous oestrogen-progestogen therapy and sequential oestrogen-progestogen therapy with each other and placebo administered over a minimum treatment period of six months. Trials had to assess which regimen was the most protective against the development of endometrial hyperplasia/carcinoma and/or caused the lowest rate of irregular bleeding. DATA COLLECTION AND ANALYSIS: Twenty three RCTs were identified and five were excluded. The reviewers assessed the eighteen included studies for quality, extracted the data independently and odds ratios for dichotomous outcomes were estimated. Outcomes analysed included frequency of endometrial hyperplasia or carcinoma, frequency of irregular bleeding and unscheduled biopsies or dilation and curettage, and adherence to therapy. MAIN RESULTS: Unopposed moderate or high dose oestrogen therapy was associated with a significant increase in rates of endometrial hyperplasia with increasing rates at longer duration of treatment and follow up. Odds ratios ranged from 5.4 (1. 4 20.9) for 6 months of treatment to 16.0 (9.3-27.5) for 36 months of treatment with moderate dose oestrogen (in the PEPI trial, 62% of those who took moderate dose oestrogen had some form of hyperplasia at 36 months compared to 2% of those who took placebo). Irregular bleeding and non adherence to treatment were also significantly more likely under these unopposed oestrogen regimens with greater effects with higher dose therapy. There was no evidence of increased hyperplasia rates, however, with low dose oestrogen. The addition of progestogens, either in continuous combined or sequential regimens, helped to prevent the development of endometrial hyperplasia and improved adherence to therapy (odds ratios of 3.7 for sequential therapy and 6.0 for continuous therapy). Irregular bleeding, however, was more likely under a continuous than a sequential oestrogen-progestogen regimen (OR = 2.3, 95% CI 2.1-2.5) but at longer duration of treatment, continuous therapy was more protective than sequential therapy in preventing endometrial hyperplasia (OR = 0.3, 95% CI 0.1-0.97). There was evidence of a higher incidence of hyperplasia under long cycle sequential therapy (progestogen given every 3 months) compared to monthly sequential therapy (progestogen given every month). No increase in endometrial cancer was seen in any of the treatment groups during the limited duration (maximum of 3 years) of these trials. (ABSTRACT TRUNCATED) PMID- 10796716 TI - Oral calorie supplements for cystic fibrosis. AB - BACKGROUND: Poor nutrition occurs frequently in children and adults with cystic fibrosis and is associated with a number of other adverse outcomes. Oral calorie supplements are used to try and increase total daily calorie intake and improve weight gain. However, they are expensive and there are concerns that they may lead to a reduction in the amount of food eaten and no overall improvement in energy intake. OBJECTIVES: To examine the evidence that in patients with cystic fibrosis, oral calorie supplements increase daily calorie intake, improve overall nutritional intake, nutritional indices, lung function, survival and quality of life. To assess possible adverse effects associated with use of oral calorie supplements. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. The companies which market oral calorie supplements were also contacted. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing use of oral calorie supplements for at least one month to increase calorie intake with no specific intervention or additional nutritional advice in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: The following outcomes were assessed: indices of nutrition and growth, anthropometric measures of body composition, calorie intake (total, from oral calorie supplements and from food), nutrient intake, eating behaviour, quality of life, specific adverse effects, lung function and mortality. MAIN RESULTS: Two trials which reported results from a total of 29 patients were suitable for inclusion in the review. From the data provided in the published reports only one item (change in weight) could be extracted from one trial for inclusion in the review. This showed no difference between intervention and comparison group. REVIEWER'S CONCLUSIONS: Oral calorie supplements are very widely used around the world in an attempt to improve nutritional status in patients with cystic fibrosis, at some considerable cost. It is therefore very disappointing that their effectiveness has not been assessed by adequate clinical trials. No conclusions can be made about the use of oral calorie supplements in cystic fibrosis from the information currently available and clinicians must balance potential benefits against possible adverse effects of treatment in making decisions about individual patients. This systematic review has clearly identified the need for a well designed, adequately-powered, multicentre, randomised controlled trial assessing the effectiveness and possible adverse effects of oral calorie supplements in cystic fibrosis. PMID- 10796717 TI - Oral steroids for cystic fibrosis. AB - BACKGROUND: In cystic fibrosis, airway obstruction and recurrent respiratory infection leads to inflammation and eventually long term lung damage, (bronchiectasis), respiratory failure and death. Inflammation occurs early in the disease process, hence the rationale for the use of anti-inflammatory agents such as oral steroids. OBJECTIVES: To assess the effectiveness of oral steroids in management of respiratory complications cystic fibrosis with particular regard to lung function and occurrence of adverse events. We aimed to to examine short term use for a respiratory exacerbation separately (up to 30 days) from long term anti inflammatory use (greater than 30 days). SEARCH STRATEGY: We searched The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised trials comparing oral corticosteroids given for a period of five to 30 days for treatment of an exacerbation or for more than 30 days used long term, with placebo or no additional therapy in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality. MAIN RESULTS: Three trials were identified studying a total of 354 patients. Two of these were long term trials with four year follow up whilst one had follow up to 12 weeks only. There was a lack of data on our predefined outcomes with common outcomes examined at different time-points and also variations in the presentation of common outcomes. A meta-analysis was not possible. Oral corticosteroids at a prednisolone equivalent dose of 1 mg/kg alternate days appear to slow the progression of lung disease in cystic fibrosis. At 24 months from commencement, 70.4% patients treated with 1mg/kg prednisolone on alternate days had an increase in per cent predicted forced vital capacity (FVC) compared to 41.6% patients treated with placebo. The mean absolute change in per cent predicted forced expiratory volume at one second (FEV1) 48 months from commencement was -2% in the 1 mg/kg alternate days prednisolone group but -6% in the placebo group. In the long term, this benefit needs to be weighed against the occurrence of adverse events. Linear growth retardation was observed as early as six months from start of treatment in the 2 mg/kg alternate days prednisolone group and from 24 months of treatment in the 1 mg/kg alternate days prednisolone group. Occurrence of adverse events, particularly glucose abnormalities, cataracts and growth retardation resulted in early termination of one of the four year studies ( approximately approximately Eigen 1995 approximately approximately ), with the group taking 2 mg/kg prednisolone on alternate days being stopped first but followed by the 1 mg/kg alternate days. REVIEWER'S CONCLUSIONS: Oral corticosteroids at a prednisolone equivalent dose of 1-2 mg/kg alternate days appear to slow the progression of lung disease in CF but this benefit needs to be weighed against the occurrence of adverse events, in particular, development of cataracts and effect on linear growth. A risk/benefit analysis of low-dose alternate days corticosteroids would be important and the role of short term use of oral steroids should be more fully evaluated. PMID- 10796718 TI - Pulsatile luteinising hormone releasing hormone for ovulation induction in subfertility associated with polycystic ovary syndrome. AB - BACKGROUND: In normal menstrual cycles, gonadotrophin releasing hormone (GnRH) secretion is pulsatile, with intervals of 60-120 minutes in the follicular phase. Treatment with pulsatile GnRH infusion by the intra-venous or subcutaneous route using a portable pump has been used successfully in patients with hypogonadotrophic hypogonadism. Assuming that the results would be similar in polycystic ovary syndrome (PCOS), pulsatile GnRH has been used to induce ovulation in patients with PCOS. But, although ovulation and pregnancy has been achieved, the use of pulsatile GnRH in PCOS patients is controversial. OBJECTIVES: To assess the effectiveness of pulsatile GnRH administration in women with clomiphene-resistant polycystic ovary syndrome (PCOS), in terms of ovulation induction, pregnancy, miscarriage, multiple pregnancy and ovarian hyperstimulation syndrome (OHSS). SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility review group was used to identify all relevant trials. Please see Review Group details. SELECTION CRITERIA: All relevant published RCTs were selected. Non-randomised controlled trials were eligible for inclusion if treatment consisted of GnRH administration versus another treatment to induce ovulation in subfertile women with PCOS. DATA COLLECTION AND ANALYSIS: A computerised MEDLINE and EMBASE search was used to identify randomised and non randomised controlled trials. The reference lists of all studies found were checked for relevant articles. One RCT (Bringer 1985a) and one abstract (Coelingh 1983) were identified this way. Relevant data were extracted independently by two reviewers (NB, MW). Validity was assessed in terms of method of randomization, completeness of follow-up, presence or absence of cross-over and co-intervention. All trials were screened and analysed for predetermined quality criteria. DATA SYNTHESIS: 2X2 tables were generated for all the relevant outcomes. Odds ratios were generated using the Peto modified Mantel Haenszel technique. MAIN RESULTS: Three RCTs and one non-randomised comparative trial were identified comparing four different treatments: GnRH versus HMG, GnRH following GnRHa pre-treatment versus no pre-treatment, GnRH and FSH versus FSH, and GnRH following GnRHa pre-treatment versus GnRH following oral contraceptive pre-treatment. This means that there was only one trial in any one comparison. In the first two studies, data of pre- and post-cross-over were not described separately. Therefore, these results could not be included in the MetaView analysis. The odds ratio for ovulation rate was 16 (95 % CI: 1.1-239) in the study comparing GnRH and FSH with FSH. When GnRH after GnRHa pre-treatment was compared with GnRH after oral contraceptive pre-treatment, an odds ratio of 7.5 (95 % CI: 1.2-46) was obtained. All trials were small and of too short duration to show any significance in pregnancy results. Per study only one to four pregnancies occurred. Multiple pregnancies were not seen. OHSS was seen only in the patients stimulated with HMG. REVIEWER'S CONCLUSIONS: The four trials describing four different comparisons with a short follow up (1 to 3 cycles) were too small to either prove or discard the value of pulsatile GnRH treatment in patients with polycystic ovary syndrome. PMID- 10796719 TI - St John's wort for depression. AB - BACKGROUND: Extracts of the plant Hypericum perforatum L. (popularly called St. John's wort) have been used in folk medicine for a long time for a range of indications including depressive disorders. OBJECTIVES: To investigate whether extracts of Hypericum are more effective than placebo and as effective as standard antidepressants in the treatment of depressive disorders in adults; and whether they have have less side effects than standard antidepressant drugs. SEARCH STRATEGY: Trials were searched in computerized general (Medline, Embase, Psychlit, Psychindex) and specialized databases (Cochrane Complementary Medicine Field, Cochrane Depression & Neurosis CRG, Phytodok); by checking bibliographies of pertinent articles; and by contacting manufacturers and researchers. SELECTION CRITERIA: Trials were included if they: (1) were randomized; (2) included patients with depressive disorders; (3) compared preparations of St. John's wort (alone or in combination with other plant extracts) with placebo or other antidepressants; and (4) included clinical outcomes such as scales assessing depressive symptoms. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, outcomes and results was extracted by at least two independent reviewers using a standard form. The main outcome measure for comparing the effectiveness of Hypericum with placebo and standard antidepressants was the responder rate ratio (responder rate in treatment group/responder rate in control group). The main outcome measure for side effects was the number of patients reporting side effects. MAIN RESULTS: 27 trials including a total of 2291 patients met inclusion criteria. 17 trials with 1168 patients were placebo controlled (16 addressed single preparations, one a combination with four other plant extracts). Ten trials (eight single preparations, two combinations of hypericum and valeriana) with 1123 patients compared hypericum with other antidepressant or sedative drugs. Most trials were four to six weeks long. Participants usually had "neurotic depression" or "mild to moderate severe depressive disorders." Hypericum preparations were significantly superior to placebo (rate ratio 2.47; 95% confidence interval 1.69 to 3.61) and similarly effective as standard antidepressants (single preparations 1.01; 0.87 to 1.16, combinations 1.52; 0.78 to 2.94). The proportions of patients reporting side effects were 26.3% for hypericum single preparations vs. 44.7% for standard antidepressants (0.57; 0.47 to 0.69), and 14. 6% for combinations vs. 26.5% with amitriptyline or desipramine (0. 49; 0.23 to 1.04). REVIEWER'S CONCLUSIONS: There is evidence that extracts of hypericum are more effective than placebo for the short-term treatment of mild to moderately severe depressive disorders. The current evidence is inadequate to establish whether hypericum is as effective as other antidepressants. Further studies comparing hypericum with standard antidepressants in well defined groups of patients over longer observations periods, investigating long term side effects, and comparing different extracts and doses are needed. PMID- 10796720 TI - Interventions for chronic suppurative otitis media. AB - BACKGROUND: Chronic suppurative otitis media (CSOM) is a serious bacterial infection of the middle ear that can follow untreated acute otitis media. OBJECTIVES: To assess the effects of different treatments for CSOM. SEARCH STRATEGY: We searched Medline from 1966 to 1996 and a bibliographic collection of the Hearing Impairment Research Group in Liverpool, UK. We handsearched two otolaryngology journals and contacted members of an international hearing network. SELECTION CRITERIA: Randomized trials of any method of management for patients with eardrum perforation and persistent otorrhea. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed eligibility and trial quality. One reviewer extracted data. We contacted investigators for clarifications. MAIN RESULTS: Twenty-four trials involving 1660 people were included. Clinical definitions and severity of CSOM varied, methodological quality was generally low and follow-up was short. Treatment with antibiotics or antiseptics accompanied by aural toilet was more effective in resolving otorrhea than no treatment (two trials, odds ratio 0.37, 95% confidence interval 0.24 to 0. 57) or aural toilet alone (six trials, odds ratio 0.31, 95% confidence interval 0.23 to 0.43). Topical treatment with antibiotics or antiseptics was more effective than systemic antibiotics (six trials, odds ratio 0.46, 95% confidence interval 0.30 to 0.69). Combining topical and systemic antibiotics was not more effective than topical antibiotics. Topical quinolones were more effective than non-quinolones (five trials, odds ratio 0.26, 95% confidence interval 0.16 to 0.41). No difference in the effectiveness of topical antibiotics and topical antiseptics was found (three studies, odds ratio 1.34, 95% confidence interval 0.64 to 2.81). Some topical antibiotic combinations may be more effective than others in resolving otorrhea. Rates of adverse drug events were low and equal between groups. REVIEWER'S CONCLUSIONS: Treatment of CSOM with aural toilet and topical antibiotics, particularly quinolones, is effective in resolving otorrhea and eradicating bacteria from the middle ear. Longterm outcomes such as preventing recurrences, closure of tympanic perforation and hearing improvement need to be further evaluated. PMID- 10796721 TI - Dieting to reduce body weight for controlling hypertension in adults. AB - OBJECTIVES: Evaluate whether weight-loss diets are more effective than regular diets or other antihypertensive therapies in controlling blood pressure and preventing morbidity and mortality in hypertensive adults. SEARCH STRATEGY: MEDLINE and The Cochrane Library were searched through November 1997. Trials known to experts in the field were included through June 1998. SELECTION CRITERIA: For inclusion in the review, trials were required to meet each of the following criteria: 1) randomized controlled trials with one group assigned to a weight-loss diet and the other group assigned to either normal diet or antihypertensive therapy; 2) ambulatory adults with a mean blood pressure of at least 140 mm Hg systolic and/or 90 mm Hg diastolic; 3) active intervention consisting of a calorie-restricted diet intended to produce weight loss (excluded studies simultaneously implementing multiple lifestyle interventions where the effects of weight loss could not be disaggregated); and 4) outcome measures included weight loss and blood pressure. DATA COLLECTION AND ANALYSIS: Studies were dual abstracted by two independent reviewers using a standardized form designed specifically for this review. The primary mode of analysis was qualitative; graphs of effect sizes for individual studies were also used. MAIN RESULTS: Eighteen trials were found. Only one small study of inadequate power reported morbidity and mortality outcomes. None addressed quality of life or general well being issues. In general, participants assigned to weight-reduction groups lost weight compared to control groups. Six trials involving 361 participants assessed a weight-reducing diet versus a normal diet. The data suggested weight loss in the range of 4% to 8% of body weight was associated with a decrease in blood pressure in the range of 3 mm Hg systolic and diastolic. Three trials involving 363 participants assessed a weight-reducing diet versus treatment with antihypertensive medications. These suggested that a stepped-care approach with antihypertensive medications produced greater decreases in blood pressure (in the range of 6/5 mm Hg systolic/diastolic) than did a weight-loss diet. Trials that allowed adjustment of participants' antihypertensive regimens suggested that patients required less intensive antihypertensive drug therapy if they followed a weight-reducing diet. Data was insufficient to determine the relative efficacy of weight-reduction versus changes in sodium or potassium intake or exercise. REVIEWER'S CONCLUSIONS: Weight-reducing diets in overweight hypertensive persons can affect modest weight loss in the range of 3-9% of body weight and are probably associated with modest blood pressure decreases of roughly 3 mm Hg systolic and diastolic. Weight-reducing diets may decrease dosage requirements of persons taking antihypertensive medications. PMID- 10796722 TI - Ovarian ablation for early breast cancer. AB - OBJECTIVES: Among women with early breast cancer, the effects of ovarian ablation on recurrence and death have been assessed by several randomised trials that now have long follow-up. In this report, the Early Breast Cancer Trialists' Collaborative Group present their third 5-yearly systematic overview (meta analysis), now with 15 years' follow-up. SEARCH STRATEGY: Trial identification procedures for the EBCTCG overviews have been described elsewhere. See under "EBCTCG" in the Breast Cancer Collaborative Review Group module. SELECTION CRITERIA: All properly randomised trials that began recruiting before 1990 which compared the ablation or suppression of ovarian function, sometimes with the addition of prednisone, versus no such adjuvant treatment for women with operable breast cancer. In practice, all the trials that can be reviewed here began before 1980, and all involved surgical or therapeutic ablation. DATA COLLECTION AND ANALYSIS: In 1995, information was sought on each patient in any randomised trial of ovarian ablation or suppression versus control that began before 1990. Data were obtained for 12 of the 13 studies that assessed ovarian ablation by irradiation or surgery, all of which began before 1980, but not for the four studies that assessed ovarian suppression by drugs, all of which began after 1985. Menopausal status was not consistently defined across trials; therefore, the main analyses are limited to women aged under 50 (rather than "premenopausal") when randomised. Oestrogen receptors were measured only in the trials of ablation plus cytotoxic chemotherapy versus the same chemotherapy alone. MAIN RESULTS: Among 2102 women aged under 50 when randomised, most of whom would have been premenopausal at diagnosis, 1130 deaths and an additional 153 recurrences were reported. 15-year survival was highly significantly improved among those allocated ovarian ablation (52.4 vs 46.1%, 6.3 [SD 2.3] fewer deaths per 100 women, logrank 2p=0.001), as was recurrence-free survival (45.0 vs 39.0%, 2p=0.0007). The numbers of events were too small for any subgroup analyses to be reliable. The benefit was, however, significant both for those with ("node positive") and for those without ("node negative") axillary spread when diagnosed. In the trials of ablation plus cytotoxic chemotherapy versus the same chemotherapy alone, the benefit appeared smaller (even for women with oestrogen receptors detected on the primary tumour) than in the trials in the absence of chemotherapy (where the observed survival improvements were about six per 100 node-negative women and 12 per 100 node-positive women). Among 1354 women aged 50 or over when randomised, most of whom would have been perimenopausal or postmenopausal, there was only a non-significant improvement in survival and recurrence-free survival. REVIEWER'S CONCLUSIONS: In women aged under 50 with early breast cancer, ablation of functioning ovaries significantly improves long term survival, at least in the absence of chemotherapy. Further randomised evidence is needed on the additional effects of ovarian ablation in the presence of other adjuvant treatments, and to assess the relevance of hormone-receptor measurements. PMID- 10796723 TI - Pre-operative GnRH analogue therapy before hysterectomy or myomectomy for uterine fibroids. AB - BACKGROUND: Uterine fibroids, smooth muscle tumours of the uterus, are found in at least 25 to 35% of women over the age of 35 years. Although some of these tumours are asymptomatic, up to 50% cause symptoms severe enough to warrant therapy and surgery is the standard treatment. Fibroid growth is stimulated by oestrogen and gonadotropin releasing hormone agonists (GnRHa) which induce a state of hypoestrogenism have been investigated as a potential treatment. GnRHa treatment causes fibroids to shrink but cannot be used long term because of unacceptable symptoms and bone loss. Therefore, GnRHa may be useful pre operatively both to reduce fibroid and uterine volume and control bleeding. OBJECTIVES: The objective of this review is to evaluate the role of pre-treatment with gonadotropin releasing hormone (GnRH) analogues prior to a major surgical procedure, either hysterectomy or myomectomy, for uterine fibroids. SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Register of Trials, MEDLINE, EMBASE, PsychLIT, Current Contents, Biological Abstracts, Social Sciences Index and CINAHL were performed. Attempts were also made to identify trials from citation lists of review articles. In most cases, the first author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of GnRH analogue treatment versus placebo, no treatment, or other medical therapy prior to surgery, either myomectomy or hysterectomy, for uterine fibroids. DATA COLLECTION AND ANALYSIS: Nineteen RCTs were identified that fulfilled the inclusion criteria for this review. The reviewers extracted the data independently and odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data of twelve trials where GnRH analogue treatment was compared with no pre-treatment and five trials where GnRH analogue treatment was compared with placebo (two trials are awaiting assessment). No RCTs of GnRH analogue treatment versus other medical therapy were identified. Results from pre operative outcomes were combined for both types of surgery but results from intra and post-operative outcomes were reported separately for myomectomy and hysterectomy. Subgroup analysis was performed according to type of control group, no pre-treatment or placebo, and for some outcomes there were additional subgroup analyses according to size of the uterus in gestational weeks. MAIN RESULTS: Pre- and post-operative haemoglobin (Hb) and haematocrit (HCT) were significantly improved by GnRH analogue therapy prior to surgery, and uterine volume, uterine gestational size and fibroid volume were all reduced. Pelvic symptoms were also reduced but some adverse events were more likely during GnRH analogue therapy. Hysterectomy appeared to be easier after pre-treatment with GnRH analogue therapy; there was reduced operating time and a greater proportion of hysterectomy patients were able to have a vaginal rather than an abdominal procedure. Duration of hospital stay was also reduced. Blood loss and rate of vertical incisions were reduced for both myomectomy and hysterectomy. Evidence of increased risk of fibroid recurrence after GnRH analogue pre-treatment in myomectomy patients was equivocal and no data were available to assess change in post-operative fertility. The increased costs associated with GnRH analogue therapy were not assessed. REVIEWER'S CONCLUSIONS: The use of GnRH analogues for 3 to 4 months prior to fibroid surgery reduce both uterine volume and fibroid size. They are beneficial in the correction of pre-operative iron deficiency anaemia, if present, and reduce intra-operative blood loss. If uterine size is such that a mid-line incision is planned, this can be avoided in many women with the use of GnRH analogues. For patients undergoing hysterectomy, a vaginal procedure is more like PMID- 10796724 TI - Brief psychological interventions ("debriefing") for trauma-related symptoms and the prevention of post traumatic stress disorder. AB - OBJECTIVES: To assess the effectiveness of brief psychological debriefing for the management of psychological distress after trauma, and the prevention of post traumatic stress disorder. SEARCH STRATEGY: Electronic searching of MEDLINE, EMBASE, PsychLit, PILOTS, Biosis, Pascal, Occ.Safety and Health, CDSR and the Trials Register of the Depression, Anxiety and Neurosis group. Hand search of Journal of Traumatic Stress. Contact with leading researchers. SELECTION CRITERIA: The inclusion criteria for all randomized studies was that they should focus on persons recently (one month or less) exposed to a traumatic event, should consist of a single session only, and that the intervention involve some form of emotional processing/ventilation by encouraging recollection/reworking of the traumatic event accompanied by normalisation of emotional reaction to the event. DATA COLLECTION AND ANALYSIS: 8 trials fulfilled the inclusion criteria. Quality was generally poor. Data from two trials could not be synthesised. MAIN RESULTS: Single session individual debriefing did not reduce psychological distress nor prevent the onset of post traumatic stress disorder (PTSD). Those who received the intervention showed no significant short term (3-5 months) in the risk of PTSD (pooled odds ratio 1.0, 95% ci 0.6-1.8). At one year one trial reported that there was a significantly increased risk of PTSD in those receiving debriefing (odds ratio 2.9, 95% ci 1.1-7.5). The pooled odds ratio for the two trials with follow ups just included unity (odds ratio 2.0, 95% ci 0.9-4.5). There was also no evidence that debriefing reduced general psychological morbidity, depression or anxiety. REVIEWER'S CONCLUSIONS: There is no current evidence that psychological debriefing is a useful treatment for the prevention of post traumatic stress disorder after traumatic incidents. Compulsory debriefing of victims of trauma should cease. PMID- 10796725 TI - Psychotherapy for bulimia nervosa and binging. AB - OBJECTIVES: The review aims to evaluate the psychotherapeutic treatments for those with binge eating syndromes, that have been tested in randomised controlled trials. Specifically, cognitive-behavioural(CBT) therapy is compared with waiting list or a non-treatment group, any other psychotherapy, CBT in a "pure self-help" form and CBT augmented by exposure and response therapy. As well, the reveiw aims to evaluate the evidence for the efficacy of other psychotherapies when compared to a no treatment control group and to evaluate the evidence for the efficacy of other psychotherapies when compared to a 'placebo' therapy. SEARCH STRATEGY: Handsearch of The International Journal of Eating Disorders since its first issue; database searches of MEDLINE, EXTRAMED, EMBASE, PSYCHLIT, CURRENT CONTENTS, LILACS, SCISEARCH, The Cochrane Collaboration Controlled Trials Register and the Cochrane Depression, Anxiety and Neurosis Group Database of Trials; citation list searching and personal approaches to authors communication are used. SELECTION CRITERIA: All studies that have tested any form of psychotherapy for adult patients with non-purging bulimia nervosa, binge eating disorder and/or EDNOS of a bulimic type, and which have applied a randomised controlled and standardized outcome methodology, are sought for the purpose of this review. DATA COLLECTION AND ANALYSIS: Data are entered into a spreadsheet programme, and into the REVMAN analysis program. Relative risk analyses are conducted of binary outcome data. The relative risk analysis is used rather than the odds ratio as the outcome measures proposed are not measuring a rare event (such as death) and the total number of studies is small. Standardized mean difference analyses are conducted of continuous variable outcome data, as the continuous outcome measures are not consistent across studies. Sensitivity analyses are conducted of a number of measures of trial quality. Data were not reported in such a way to do subgroup analyses, but the effect of treatment on depressive symptoms, psychosocial and/or interpersonal functioning, general psychiatric symptoms and weight is examined where possible. Chi-square tests for homogeneity are done, @ 5% level of significance, using a fixed effects model. Funnel plots to evaluate presence of publication bias are completed and available in a text file upon request. MAIN RESULTS: To date, 1360 trials have been generated by searching and 58 trials have been evaluated in detail. Because of a relatively high number of exclusions (n=12) the trial inclusion criteria were broadened to include those with non-blinded outcome assessment, providing 20 trials for analyses. Because of incomplete published and available data, at best up to 10 studies had data available for any single analysis. The maximum number of total patients included in a single analysis is 396. The majority of studies (18) evaluate patients with bulimia nervosa of a purging type. CBT is superior to waiting list controls with respect to abstinence from binge eating (RR 0.64 CI.53 .78). CBT is not superior to other psychotherapies with respect to abstinence from binge eating (RR.79, CI.54-1.17). CBT in a full or less intensive form is not significantly superior to CBT in a pure self-help form. Augmentation of CBT with exposure therapy is not more effective than CBT alone. NonCBT psychotherapies also have significantly greater abstinence rates in comparisons with wait-list controls, but there is a paucity of such studies (RR 0.67, CI.56 .81, n=3 studies). Funnel plots suggest a bias towards publication of positive outcome studies only. REVIEWER'S CONCLUSIONS: There is small body of evidence for the efficacy of cognitive-behaviour therapy in bulimia nervosa and similar syndromes, but the quality of trials is very variable (e.g. the majority, 12, are not blinded) and sample sizes are often very small. More trials are needed, particularly for binge eating disorder and other EDNOS syndromes, and evalu PMID- 10796726 TI - Day care for pre-school children. AB - BACKGROUND: The debate about how, where and by whom young children should be looked after is one which has occupied much social policy and media attention in recent years. Mothers undertake most of the care of young children. Internationally, out-of-home day-care provision ranges widely. These different levels of provision are not simply a response to different levels of demand for day-care, but reflect cultural and economic interests concerning the welfare of children, the need to promote mothers' participation in paid work, and the importance of socialising children into society's values. At a time when a decline in family values is held responsible for a range of social problems, the day-care debate has a special prominence. OBJECTIVES: To quantify the effects of out-of-home day-care for preschool children on educational, health and welfare outcomes for children and their families. SEARCH STRATEGY: Randomised controlled trials of day-care for pre-school children were identified using electronic databases, hand searches of relevant literature, and contact with authors. SELECTION CRITERIA: Studies were included in the review if the intervention involved the provision of non-parental day care for children under 5 years of age, and the evaluation design was that of a randomised or quasi-randomised controlled trial. DATA COLLECTION AND ANALYSIS: A total of eight trials were identified after examining 920 abstracts and 19 books. The trials were assessed for methodological quality. MAIN RESULTS: Day-care increases children's IQ, and has beneficial effects on behavioural development and school achievement. Long term follow up demonstrates increased employment, lower teenage pregnancy rates, higher socio-economic status and decreased criminal behaviour. There are positive effects on mothers' education, employment and interaction with children. Effects on fathers have not been examined. Few studies look at a range of outcomes spanning the health, education and welfare domains. Most of the trials combined non-parental day-care with some element of parent training or education (mostly targeted at mothers); they did not disentangle the possible effects of these two interventions. The trials had other significant methodological weaknesses, pointing to the importance of improving on study design in this field. All the trials were carried out in the USA. REVIEWER'S CONCLUSIONS: Day care has beneficial effect on children's development, school success and adult life patterns. To date, all randomised trials have been conducted among disadvantaged populations in the USA. The extent to which the results are generaliseable to other cultures and socioeconomic groups has yet to be evaluated. PMID- 10796727 TI - Calcium channel blockers for acute traumatic brain injury. AB - BACKGROUND: Acute traumatic brain injury is a major cause of death and disability. Calcium channel blockers have been used in an attempt to prevent cerebral vasospasm after injury, maintain blood flow to the brain and so prevent further damage. OBJECTIVES: To estimate the effects of calcium channel blockers in patients with acute traumatic brain injury and in a subgroup of brain injury patients with traumatic subarachnoid haemorrhage. SEARCH STRATEGY: Hand searching and electronic searching for randomized controlled trials available by October 1997. SELECTION CRITERIA: Randomized controlled trials in patients with all levels of severity of clinically diagnosed acute traumatic brain injury. DATA COLLECTION AND ANALYSIS: Two reviewers (JL and CG) independently assessed the identified studies for eligibility and extracted data from each study. Summary odds ratios were calculated using the Mantel-Haenszel method. MAIN RESULTS: Four RCTs were identified as eligible for inclusion in the systematic review. The effect of calcium channel blockers on the risk of death was reported in all RCTs. The pooled odds ratio for the four studies was 0.91 (95% confidence interval 0. 70 to 1.17). For the three RCTs that reported death and severe disability the pooled odds ratio was 0.85 (95% CI 0.68 to 1.07). In the two RCTs which reported the risk of death in a sub group of traumatic subarachnoid haemorrhage patients, the pooled odds ratio was 0.59 (95% CI 0.37 to 0.94). Three RCTs reported death and severe disability as an outcome in this subgroup, and the pooled odds ratio was 0.67 (95% CI 0.46 to 0.98). REVIEWER'S CONCLUSIONS: This systematic review of randomized controlled trials of calcium channel blockers in acute traumatic head injury patients shows that considerable uncertainty remains over their effects. The effect of nimodipine in a subgroup of brain injury patients with subarachnoid haemorrhage shows a beneficial effect, though the increase in adverse reactions suffered by the intervention group may mean that the drug is harmful for some patients. PMID- 10796728 TI - Hyperventilation therapy for acute traumatic brain injury. AB - BACKGROUND: Because hyperventilation is often associated with a rapid fall in intracranial pressure, it has been assumed to be effective in the treatment of severe head injury. Hyperventilation reduces raised intracranial pressure by causing cerebral vasoconstriction and a reduction in cerebral blood flow. Whether reduced cerebral blood flow improves neurological outcome however, is unclear. OBJECTIVES: To quantify the effect of hyperventilation on death and neurological disability following head injury. SEARCH STRATEGY: The search strategy drew on that of the Injuries Group as a whole. The reference lists of all relevant articles identified were checked and the first author of reports was contacted to ask for assistance in identifying any further trials. Most recent search was done in September 1999. SELECTION CRITERIA: All randomised trials of hyperventilation, in which study participants had a clinically defined acute traumatic head injury of any severity. There were no language restrictions. DATA COLLECTION AND ANALYSIS: We collected data on the participants, the timing and duration of the intervention, duration of follow up, neurological disability and death. Relative risks (RR) and 95% confidence intervals were calculated for each trial on an intention to treat basis. Timing, degree and duration of hyperventilation were identified a-priori as potential sources of heterogeneity between trials. MAIN RESULTS: One trial of 113 participants was identified. Hyperventilation alone, as well as in conjunction with the buffer THAM showed a beneficial effect on mortality at one year after injury, although the effect measure was imprecise (RR=0.73; 95% CI 0.36;1.49 and RR=0.89; 95% CI 0.47;1.72 respectively). This improvement in outcome was not supported by an improvement in neurological recovery. For hyperventilation alone, the RR for death or severe disability was 1. 14 (95% CI 0.82;1.58). The RR for death or severe disability in the hyperventilation plus THAM group, was 0.87 (95% CI 0.58;1.28). REVIEWER'S CONCLUSIONS: The data available are inadequate to assess any potential benefit or harm that might result from hyperventilation in severe head injury. Randomised controlled trials to assess the effectiveness of hyperventilation therapy following severe head injury are needed. PMID- 10796729 TI - Colloids versus crystalloids for fluid resuscitation in critically ill patients. AB - BACKGROUND: Colloid solutions are widely used in fluid resuscitation of critically ill patients. There are several choices of colloid and there is ongoing debate about the relative effectiveness of colloids compared to crystalloid fluids. OBJECTIVES: To assess the effects on mortality of colloids compared to crystalloids for fluid resuscitation in critically ill patients. SEARCH STRATEGY: We searched the Injuries Group specialised register, Cochrane Controlled Trials Register, MEDLINE, EMBASE and BIDS Index to Scientific and Technical Proceedings and checked the reference lists of trials and review articles. SELECTION CRITERIA: All randomised and quasi-random trials of colloids compared to crystalloids, in patients requiring volume replacement. Cross-over trials and trials in pregnant women and neonates were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and rated quality of allocation concealment. Trials with a 'double-intervention' such as those, which compared colloid in hypertonic crystalloid to isotonic crystalloid, were analysed separately. The analysis was stratified according to colloid type and quality of allocation concealment. MAIN RESULTS: Colloids compared to crystalloids: Albumin or plasma protein fraction: Eighteen trials reported data on mortality, including a total of 641 patients. The pooled relative risk from these trials was 1.52 (95% confidence interval 1.08 to 2.13). The risk of death in the albumin treated group was 6% higher than in the crystalloid treated group (1% to 11%). When the trial with poor quality allocation concealment was excluded the pooled relative risk was 1.34 (0.95 to 1.89). Hydroxyethylstarch: Seven trials compared hydroxyethylstarch with crystalloids including a total of 197 randomised participants. The pooled relative risk was 1.16 (0.68 to 1.96). Modified gelatin: Four trials compared modified gelatin with crystalloid including a total of 95 randomised participants. The pooled relative risk was 0.50 (0. 08 to 3.03). Dextran: Eight trials compared dextran with a crystalloid including a total of 668 randomised participants. The pooled relative risk was 1.24 (0.94 to 1.65). Colloids in hypertonic crystalloid compared to isotonic crystalloid: Eight trials compared dextran in hypertonic crystalloid with isotonic crystalloid, including 1283 randomised participants. The pooled relative risk was 0.88 (0.74 to 1.05). REVIEWER'S CONCLUSIONS: There is no evidence from randomised controlled trials that resuscitation with colloids reduces the risk of death compared to crystalloids in patients with trauma, burns and following surgery. As colloids are not associated with an improvement in survival, and as they are more expensive than crystalloids, it is hard to see how their continued use in these patient types can be justified outside the context of randomised controlled trials. PMID- 10796730 TI - Oestrogen and progestogen hormone replacement therapy for peri-menopausal and post-menopausal women: weight and body fat distribution. AB - BACKGROUND: Hormone replacement therapy (HRT) is commonly prescribed to treat menopausal symptoms and to prevent post-menopausal bone loss. However, many women are concerned about hormonal replacement therapy because they believe that such treatment will result in weight gain. The effect of HRT on weight and body fat distribution has not yet been examined in systematic reviews. It is an important topic since many women decline oestrogen therapy due to their concerns about resultant weight gain, and thus forego its potential therapeutic benefits. OBJECTIVES: To evaluate the effect of unopposed oestrogen or combined oestrogen and progestogen hormone replacement therapy (HRT) upon the weight and body fat distribution of perimenopausal and postmenopausal women. SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility Group was used for the identification of randomised controlled trials (RCTs). Computerised searches of MEDLINE, EMBASE, Current Contents, Biological Abstracts and CINAHL were performed. Attempts were made to identify trials from citation lists of review articles and relevant papers already obtained. In most cases, first authors of each eligible trial were contacted for additional information. All those trials that had been located as at August 1998 were examined for eligibility. SELECTION CRITERIA: All randomised, placebo or no treatment controlled trials that detailed the effect of HRT on weight or body fat distribution, including studies where HRT was combined with other therapy such as diet, supplements or exercise. Studies were eligible for consideration even though the main focus of the trial may have been on another aspect of HRT. Previous HRT use should have ceased at least one month (in the case of patches, cream or gel) or three months (for oral preparations or subcutaneous pellets) before commencement of the study. DATA COLLECTION AND ANALYSIS: Twenty two RCTs were identified that fulfilled the inclusion criteria for this review. The results of one trial were not available in a form that allowed it to be included in the meta-analysis; however, it has been included in the text of the review for discussion. Twenty four RCTs are awaiting assessment pending additional information from first authors. Two reviewers extracted the data independently, and the weighted mean differences for continuous outcomes were estimated from the data. Results for unopposed oestrogen and combined oestrogen were analysed separately, and the effect of each treatment regimen on body weight, BMI, waist-hip ratio, fat mass and skinfold measurement was examined where available. The effect of differing dosage levels on these parameters was also examined. MAIN RESULTS: Outcomes were evaluated separately for unopposed oestrogen and oestrogen/progestogen regimens. Statistical analysis was performed using the weighted mean difference for continuous outcomes as recommended by the Cochrane Menstrual Disorders and Subfertility Group. No statistically significant difference was found in mean weight gain between those using unopposed oestrogen and non-HRT users (0.66 kg, 95% CI -0.62, 1.93). No significant difference was found in mean weight gain between those using oestrogen/progestogen therapy and non-HRT users (-0.47 kg, 95% CI -1.63, 0.69). Insufficient data exist to enable meta-analysis of the effect of unopposed oestrogen on BMI. The reviewers found no statistically significant difference in mean BMI increase between those using oestrogen/progestogen and non-HRT users ( 0.50, 95% CI -1.06, 0.06). Insufficient data exist to enable meta-analysis of the effect of HRT on waist-hip ratio, fat mass or skinfold thickness. REVIEWER'S CONCLUSIONS: There is evidence of no effect of unopposed oestrogen or combined oestrogen on body weight, indicating that these regimens do not cause extra weight gain in addition to that normally gained at menopause. (ABSTRACT TRUNCATED) PMID- 10796731 TI - Modern combined oral contraceptives for pain associated with endometriosis. AB - BACKGROUND: Endometriosis is a major women's health-care problem. It causes pain and/or infertility, and affects millions of women worldwide. Endometriosis is defined according to histological criteria by the presence of tissue resembling endometrium in sites outside the uterus, most commonly the ovaries and peritoneum. The aim of treatment has been to remove the deposits of ectopic endometrium that are thought to be responsible for the symptoms of endometriosis. This can be achieved surgically by destroying or removing the implants; medical therapies induce atrophy within the hormonally-dependent ectopic endometrium. The duration of hormonal treatment may be limited by unwanted side effects. There is some evidence, however, from epidemiological research that current use of the combined oral contraceptive pill (OCP) is associated with a reduced incidence of endometriosis. The combined pill has the great advantage over other hormonal treatments that it can be taken indefinitely. OBJECTIVES: The aim of this review was to establish the role of modern oral contraceptives in the management of painful symptoms ascribed to endometriosis. SEARCH STRATEGY: The search strategy of the Menstrual Disorders Group was utilised to identify all randomised trials of the use of oral contraceptives in the treatment of symptomatic endometriosis. In addition a search of the Cochrane Controlled Trials Register was undertaken together with approaches to pharmaceutical companies. SELECTION CRITERIA: All truly randomised controlled trials of the use of oral contraceptive pills in the treatment of women of reproductive age with symptoms ascribed to the diagnosis of endometriosis made visually at a surgical procedure, were included. DATA COLLECTION AND ANALYSIS: Study quality assessment and data extraction was carried out independently by two reviewers. One of the assessors was an expert in the content matter. MAIN RESULTS: Only one study was identified which satisfied the inclusion criteria. The oral contraceptive used in a conventional manner was less effective than a GnRH analogue in the relief of dysmenorrhoea. No significant difference was noted between the effectiveness of the oral contraceptive pill and a GnRH analogue in the relief of dyspareunia or non-menstrual pain. Headaches and weight gain were more commonly associated with oral contraceptive usage than with GnRH analogue usage whereas hot flushes, insomnia and vaginal dryness were less common. REVIEWER'S CONCLUSIONS: There is a paucity of data relating to the use of oral contraceptive preparations in the treatment of symptomtic endometriosis. The data such as it is supports the common practice of the use of the oral contraceptive pill as a first line therapy but further research is required to fully evaluate its role in the management of endometriosis. PMID- 10796732 TI - Nebulised anti-pseudomonal antibiotics for cystic fibrosis. AB - BACKGROUND: Lung damage associated with persistent infection by Pseudomonas aeruginosa is the major cause of morbidity and mortality in people with cystic fibrosis. Nebulised antibiotics are commonly used for treatment of this infection. OBJECTIVES: To examine the evidence that nebulised anti-pseudomonal antibiotic treatment in patients with cystic fibrosis reduces frequency of exacerbations of infection, improves lung function, quality of life and survival. To assess adverse effects of nebulised anti-pseudomonal antibiotic treatment. SEARCH STRATEGY: Trials were identified from the Cochrane Cystic Fibrosis and Genetic Disorders Group clinical trials register. Companies which marketed nebulised anti-pseudomonal antibiotics were contacted for information on unpublished trials. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: Trials were selected if, nebulised anti-pseudomonal antibiotics treatment was used in patients with cystic fibrosis, treatment was used for four weeks or more, allocation to treatment was randomised or pseudo-randomised, and there was a placebo or a no placebo control group. DATA COLLECTION AND ANALYSIS: References to nebulised antibiotics were retrieved from the register. Two reviewers independently selected trials from this sample according to the criteria. Each judged the quality of selected trials. One reviewer extracted data from these trials. MAIN RESULTS: Ten trials with 758 participants were included in the review. Sixty eight % of participants were in one trial. Duration of trials ranged from one to 32 months. There was large variation in study design and outcome measures which limited meta-analysis. Lung function measured as forced expired volume in one second (FEV1) was generally better in the treated groups than in control groups but a pooled estimate of effect was not possible. In the largest trial of 520 participants the FEV1 was 11.9% predicted (95% confidence interval 8.1to 15.6) higher in the treated group than in the placebo control group after 5 months. In three trials with 581 participants the number of participants having at least one hospital admission during a trial was reduced in the groups treated with nebulised antibiotics, odds ratio 0.69 (95% confidence Interval 0.50 to 0.96). In two trials with 591 participants the number of participants having and at least one course of antibiotic therapy was reduced in the treated group, odds ratio 0.62 (95% confidence interval 0.45 to 0.87). There was no evidence of renal or auditory toxicity. Resistance to antibiotic increased more in the antibiotic treated group than in placebo group. The most evidence for a single drug is for tobramycin, which was used in total daily doses from 40mg to 1800mg. REVIEWER'S CONCLUSIONS: Nebulised anti-pseudomonal antibiotic treatment improves lung function and reduces frequency of exacerbations of infection in people with cystic fibrosis. There is a need for more evidence for effect on quality of life and survival, for longer duration trials to determine if this benefit is maintained and to determine the significance of development of antibiotic resistant organisms. PMID- 10796733 TI - Cognitive behaviour therapy for adults with chronic fatigue syndrome. AB - OBJECTIVES: 1. To systematically review all randomised controlled trials of cognitive-behaviour therapy (CBT) for adults with chronic fatigue syndrome (CFS); 2. To test the hypothesis that CBT is more effective than orthodox medical management or other interventions in adults with CFS. SEARCH STRATEGY: 1. Electronic searching of bibliographic databases, including Medline, PsycLIT, Biological Abstracts, Embase, SIGLE, Index to Theses, Index to Scientific and Technical Proceedings, and Science Citation Index, using multiple search terms in order to perform a highly sensitive search. 2. Electronic searching of the Trials Register of the Depression, Anxiety and Neurosis group. 3. Citation lists of relevant studies and reviews were perused for other relevant trials. 4. Contact with the principal authors of relevant studies, and with researchers in the field. SELECTION CRITERIA: All randomised controlled trials were included in which - adult patients with CFS; - received CBT or a control intervention, being either orthodox medical management or another intervention; - and whose outcomes were assessed in an appropriate way. CBT could be either type 'A' (encouraging return to 'normal' levels of rest and activity) or type 'B' (encouraging rest and activity which were within levels imposed by the disorder). DATA COLLECTION AND ANALYSIS: The two reviewers worked independently throughout the selection of trials and data extraction, comparing findings only when there was disagreement. Relevant trials were allocated to one of three quality categories. Full data extraction, using a standardised data extraction sheet, was performed on studies which were of high or moderate quality. Trials of low quality were excluded from the review. The comparisons made to test the review hypothesis were of type 'A' CBT versus other intervention(s), and of type 'B' CBT versus other intervention(s). Functional outcome was used as the main outcome for comparison, but other appropriate outcomes were compared where possible. Results were synthesised using the Review Manager software. For dichotomous data, the odds ratio was calculated for each study. For continuous data, effect sizes were obtained and the standardised mean difference, with 95% confidence intervals, was calculated. MAIN RESULTS: Only three relevant trials of adequate quality were found. These trials demonstrated that CBT significantly benefits physical functioning in adult out-patients with CFS when compared to orthodox medical management or relaxation. It is necessary to treat about two patients to prevent one additional unsatisfactory physical outcome about six months after treatment end. CBT appeared highly acceptable to the patients in these trials. There is no satisfactory evidence for the effectiveness of CBT in patients with the milder forms of CFS found in primary care or in patients who are so disabled that they are unable to attend out-patients. Additionally, there is no satisfactory evidence for the effectiveness of group CBT. REVIEWER'S CONCLUSIONS: Cognitive behaviour therapy appears to be an effective and acceptable treatment for adult out-patients with chronic fatigue syndrome. CFS is a common and disabling disorder. Its sufferers deserve the medical profession to be more aware of the potential of this therapy to bring lasting functional benefit, and health service managers to increase its availability. Further research is needed in this important area. Trials should conform to accepted standards of reporting and methodology. The effectiveness of CBT in more and less severely disabled patients than those usually seen in out-patient clinics needs to be assessed. Trials of group CBT and in-patient CBT compared to orthodox medical management, and of CBT compared to graded activity alone, also need to be conducted. PMID- 10796734 TI - Tamoxifen for relapse of ovarian cancer. AB - BACKGROUND: Tamoxifen is an important drug for treating breast cancer. Ovarian cancer cells are known to possess receptors for hormones and may thus also respond to tamoxifen. OBJECTIVES: Tamoxifen is used to treat breast cancer in women whose tumours have oestrogen receptors. Since ovarian cancers also commonly have oestrogen receptors, it has been suggested that tamoxifen may be of some benefit. The objective of this review was to assess the effects of tamoxifen in women with relapsed ovarian cancer. SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Group trials register and references from relevant articles. We also contacted researchers and drug companies. SELECTION CRITERIA: Randomised and non-randomised studies of tamoxifen in women with ovarian cancer who have not responded to conventional chemotherapy. Only trials involving 10 or more patients were included. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and extracted data from non-randomised studies. Two reviewers were to have independently assessed the quality and extracted data from any randomised trials found. MAIN RESULTS: Eleven non-randomised series, one non-randomised phase two study and one randomised trial were included. Only observational data from women treated with tamoxifen are reported. Fifty-nine of 568 women (10.4%) treated with tamoxifen achieved an objective response to treatment. However this varied from 0% to 56% in different studies. Stable disease, for variable periods of four weeks or more, was observed in 109 of 356 (30.6%) women from eight studies. There were not enough data to assess duration of response, survival, or the palliative effect of tamoxifen on symptom control or quality of life. REVIEWER'S CONCLUSIONS: There is some evidence from observational studies that tamoxifen may produce a response in a modest proportion of women with relapsed ovarian cancer. However, there are no reliable data from randomised controlled trials. PMID- 10796735 TI - Interventions for encouraging sexual lifestyles and behaviours intended to prevent cervical cancer. AB - BACKGROUND: Cervical cancer is one of the most common cancers affecting women world-wide. Prevention falls into two main categories - primary and secondary. Primary prevention is characterised by health promotion to promote lifestyles and behaviours minimising risk of cervical cancer. Interventions to promote the use of condoms for sexual intercourse (especially early intercourse amongst young women), sexual partner reduction, and negotiated safer sex strategies has been recommended as one approach to limit the spread of Human Papilloma Virus (HPV), one of the major risk factors for cervical cancer. OBJECTIVES: To determine the effectiveness of health education interventions to promote sexual risk reduction behaviours amongst women in order to reduce transmission of HPV. SEARCH STRATEGY: Electronic searching of EMBASE, ERIC, MEDLINE, PsycLIT, Social Science Citation Index and the CCTR were undertaken using a highly sensitive search strategy. Hand searching took place of selected journals and reference lists. SELECTION CRITERIA: Studies were included if they evaluated educational interventions targeting women only, and measured the impact on : either a behavioural outcome such as condom use for sexual intercourse, partner reduction, or abstinence; or a clinical outcome such as incidence of a sexually transmitted disease (STD). DATA COLLECTION AND ANALYSIS: Data were extracted and methodological quality was assessed independently by two reviewers and any discrepancies were resolved between them. Ten per cent of the total number of studies were reviewed additionally by a third reviewer as a quality check and differences in judgement were resolved accordingly. MAIN RESULTS: Thirty trials met the inclusion criteria for the review. All of them had the primary aim of preventing HIV and other STDs rather than cervical cancer. Four core methodological qualities were present in 10 of the 30 studies and constitute the subset from which potentially reliable conclusions may be drawn. Each of the 10 studies showed a statistically significant positive effect on sexual risk reduction, typically with increased use of condoms for vaginal intercourse. This positive effect was generally sustained up to three months after intervention. REVIEWER'S CONCLUSIONS: Educational interventions targeting socially and economically disadvantaged women in which information provision is complemented by sexual negotiation skill development can encourage at least short-term sexual risk reduction behaviour. This has the potential to reduce the transmission of HPV, thus possibly reduce the incidence of cervical carcinoma. PMID- 10796736 TI - Collection devices for obtaining cervical cytology samples. AB - OBJECTIVES: The false-negative rate of cervical smears varies between 1.5% and 55%. This variation may be partly due to differences in sampling device and technique. The objective of this review was to assess different cervical sampling devices for collecting endocervical cells, which are thought to be a surrogate for detection of abnormal cells and adequate smear rates. SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Group trials register and MEDLINE up to July 1997. We also handsearched 16 journals. SELECTION CRITERIA: Randomised and quasi-randomised trials and non-randomised comparative studies comparing cervical smear collection devices in women attending for primary screening, colposcopy following an abnormal smear or colposcopy after treatment. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted data. Study quality was assessed. MAIN RESULTS: Thirty-four trials and six observational comparative studies were included. The Ayre spatula was shown to be less effective compared with extended tip spatulas for collecting endocervical cells in eight trials (odds ratio 2.25, 95% confidence interval 2.06 to 2.44). Use of a spatula with the cytobrush was more effective than spatula alone at collecting endocervical cells (odds ratio 3.33, 95% 3.05 to 3.63) and the same effect was present for adequate smear rates (odds ratio 1.51 95% 1.19-1.92). Extended tip spatulas were also superior for the detection of dyskaryosis in seven trials (odds ratio 1.21, 95% confidence interval 1.10 to 1.33). Based on data from two trials and three observational studies, smears that contained endocervical cells were more likely to detect dyskaryosis, particularly in severe disease. The proportion of smears with endocervical cells present increased with increasing severity of the disease. REVIEWER'S CONCLUSIONS: Extended tip spatulas of various designs appear to be better for collecting endocervical cells than the commonly used Ayre spatula. The most effective combination appears to be the cytobrush with an extended tip spatula. The rate of detection of endocervical cells appears to be a valid and convenient surrogate for the ability to detect dyskaryosis and for adequate smear rates. The ability of the extended tip spatula with the cytobrush compared with the extended tip spatula alone to detect disease, needs to be evaluated in a trial. PMID- 10796737 TI - Progestagens for endometrial cancer. AB - OBJECTIVES: Progestagen therapy following primary surgery for endometrial cancer has been advocated to reduce the risk of recurrence. The objective of this review was to assess the effect of adjuvant progestagen therapy in endometrial cancer. SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Group trials register and MEDLINE up to May 1999. SELECTION CRITERIA: Randomised trials of progestagen therapy in women who have had surgery for endometrial cancer. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and two reviewers abstracted data independently. MAIN RESULTS: Six trials involving 4351 women were identified. Three trials included women with stage one disease only, whereas three included women with more advanced disease. Based on five trials, overall survival was not improved by adjuvant progestagen therapy (OR 1.05, 95% CI 0.88 to 1.24). Endometrial cancer deaths and relapse of disease appears to be reduced by progestagen therapy OR 0.88 95% CI (0.71-1.1) and 0. 81 95% CI (0.65-1.01) respectively. However, non-endometrial cancer related deaths were more common in women treated with progestagens OR 1.33 (1.02-1.73). REVIEWER'S CONCLUSIONS: Current evidence does not support the use of adjuvant progestagen therapy in the primary treatment of endometrial cancer. PMID- 10796738 TI - Allopurinol for chronic prostatitis. AB - OBJECTIVES: To determine the effects of allopurinol in the treatment of chronic prostatitis SEARCH STRATEGY: Trials were searched in computerized general and specialized databases (MEDLINE, Cochrane Library, Cochrane Prostate Group database), bibliographies of obtained articles, and direct contact with authors. SELECTION CRITERIA: All randomized trials of allopurinol versus placebo used to treat patients with chronic prostatitis. Acute prostatitis, bacterial prostatitis, and asymptomatic prostatitis were excluded. The main outcome measure was the change in patient-reported discomfort. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently for the outcomes of change in patient reported discomfort, investigator graded prostate pain, leukocyte counts, and biochemical indices. MAIN RESULTS: Only one trial with 54 men lasting 240 days (with 330 days of follow-up) met study inclusion criteria. There was a statistically significant change favoring allopurinol in patient-reported discomfort between the study and control groups at follow-up. Between days 45 225, the mean score was -0.95 (s.d. 0.19) for the allopurinol group (7 men), compared with -0.47 (s.d. 0.21) for the placebo group (7 men). The weighted mean difference (WMD) was -0.48 (95%CI -0.690, -0.270). The mean score between days 45 135 was -1.08 (s.d. 1.29) for the 25 men in the allopurinol group, compared with 0.21 (sd 0.97) for the 14 men in the control group. The WMD was -0.87 (95%CI 1.587, -0.153). The allopurinol group had significantly less investigator graded prostate pain and had lower levels of serum urate, urine urate, and expressed prostatic secretion urate and xanthine. No significant differences between the two groups regarding leukocyte counts were found. No patient receiving allopurinol had any significant side effects. Three patients in the placebo group dropped out because of side effects. REVIEWER'S CONCLUSIONS: One small trial of allopurinol for treating chronic prostatitis showed improvements in patient reported symptom improvement, investigator-graded prostate pain, and biochemical parameters. However, the data provided, the measures used, and the statistics presented do not make these findings convincing that changes in urine and prostatic secretion composition regarding purine and pyrimidine bases resulted in the relief of symptoms. Further studies of allopurinol treatment using standardized and validated outcomes measures and analyses are necessary to determine whether allopurinol is effective. PMID- 10796739 TI - Cernilton for benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH), nonmalignant enlargement of the prostate, can lead to obstructive and irritative lower urinary tract symptoms (LUTS). The pharmacologic use of plants and herbs (phytotherapy) for the treatment of LUTS associated with BPH has been growing steadily. Cernilton, prepared from the rye-grass pollen Secale cereale, is one of the several phytotherapeutic agents available for the treatment of BPH. OBJECTIVES: This systematic review aims to assess the effects of Cernilton on urinary symptoms and flow measures in men with benign prostatic hyperplasia (BPH). SEARCH STRATEGY: Trials were searched in computerized general and specialized databases (MEDLINE, EMBASE, Cochrane Library, Phytodok), by checking bibliographies, and by contacting manufacturers and researchers. SELECTION CRITERIA: Trials were eligible if they were: (1) randomized controlled trials or controlled clinical trials comparing Cernilton with placebo or other BPH medications in men with BPH; and (2) included clinical outcomes such as urologic symptom scales, symptoms, or urodynamic measurements. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes was extracted by at least two independent reviewers using a standard form. Main outcome measure for comparing the effects of Cernilton with placebo and standard BPH medications were the change in urologic symptoms scales. Secondary outcomes included changes in nocturia as well as urodynamic measures (peak and mean urine flow, residual volume, prostate size). Main outcome measure for side effects was the number of men reporting side effects. MAIN RESULTS: 444 men were enrolled in 2 placebo-controlled and 2 comparative trials lasting from 12 to 24 weeks. Three studies used a double-blind method although treatment allocation concealment was unclear in all. Cernilton improved "self rated urinary symptoms" (percent reporting satisfactory or improving symptoms) versus placebo and Tadenan. The weighted risk ratio (RR) for self-rated improvement versus placebo was 2.40 [95% CI = 1.21, 4. 75], and the weighted RR versus Tadenan was 1.42 [95% CI = 1.21, 4. 75]. Cernilton reduced nocturia compared with placebo and Paraprost. Versus placebo, the weighted RR was 2.05 [95% CI = 1.41, 3.00], and versus Paraprost, the WMD was -0.40 times per evening [95% CI = -0. 73, -0.07]. Cernilton did not improve urinary flow rates, residual volume or prostate size compared to placebo or the comparative study agents. Adverse events were rare and mild. The withdrawal rate for Cernilton was 4.8% compared to 2.7% for placebo and 5.2% for Paraprost. REVIEWER'S CONCLUSIONS: The Cernilton trials analyzed were limited by short duration, limited number of enrollees, gaps in reported outcomes, and unknown quality of the preparations utilized. The comparative trials lacked a proven active control. The available evidence suggests Cernilton is well tolerated and modestly improves overall urologic symptoms including nocturia. Additional randomized placebo and active controlled trials are needed to evaluate the long-term clinical effectiveness and safety of Cernilton. PMID- 10796740 TI - Beta-sitosterols for benign prostatic hyperplasia. AB - OBJECTIVES: This systematic review aimed to assess the effects of beta sitosterols (B-sitosterol) on urinary symptoms and flow measures in men with of benign prostatic hyperplasia (BPH). SEARCH STRATEGY: Trials were searched in computerized general and specialized databases (MEDLINE, EMBASE, Cochrane Library, Phytodok), by checking bibliographies, and by contacting manufacturers and researchers. SELECTION CRITERIA: Trials were eligible for inclusion provided they (1) randomized men with BPH to receive B-sitosterol preparations in comparison to placebo or other BPH medications, and (2) included clinical outcomes such as urologic symptom scales, symptoms, or urodynamic measurements. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes was extracted by at least two independent reviewers using a standard form. Main outcome measure for comparing the effectiveness of B-sitosterols with placebo and standard BPH medications was the change in urologic symptom scale scores. Secondary outcomes included changes in nocturia as well as urodynamic measures (peak and mean urine flow, residual volume, prostate size). Main outcome measure for side effects was the number of men reporting side effects. MAIN RESULTS: 519 men from 4 randomized, placebo-controlled, double-blind trials, (lasting 4 to 26 weeks) were assessed. 3 trials used non-glucosidic B-sitosterols and one utilized a preparation that contained 100% B-sitosteryl-B-D-glucoside. B Sitosterols improved urinary symptom scores and flow measures. The weighted mean difference (WMD) for the IPSS was -4.9 IPSS points (95%CI = -6.3 to -3.5, n = 2 studies). The WMD for peak urine flow was 3.91 ml/sec (95%CI = 0.91 to 6.90, n = 4 studies) and the WMD for residual volume was -28.62 ml (95%CI = -41. 42 to 15.83, n = 4 studies). The trial using 100% B-sitosteryl-B-D-glucoside (WA184) show improvement in urinary flow measures. B-sitosterols did not reduce prostate size. Withdrawal rates for men assigned to B-sitosterol and placebo were 7.8% and 8. 0%, respectively. REVIEWER'S CONCLUSIONS: The evidence suggests non-glucosidic B-sitosterols improve urinary symptoms and flow measures. Their long term effectiveness, safety and ability to prevent BPH complications are not known. PMID- 10796741 TI - Pharmacological interventions for acute spinal cord injury. AB - BACKGROUND: Acute spinal cord injury is a devastating condition typically affecting young people with a preponderance of males. Pharmacological treatment in the early hours of the injury is aimed at reducing the extent of permanent paralysis during the rest of the patient's life. OBJECTIVES: To review randomized trials of pharmacological therapies for acute spinal cord injury. SEARCH STRATEGY: The review draws on the search strategy developed by the Cochrane Injuries Group. In addition, files of the National Acute Spinal Cord Injury Study have been reviewed. SELECTION CRITERIA: All published or unpublished randomized controlled trials of pharmacological treatment for acute spinal cord injury in any language. DATA COLLECTION AND ANALYSIS: Data have been abstracted from original trial reports. For the NASCIS, Japanese and French trials, additional data (e.g. SDs) have been obtained from the original authors. MAIN RESULTS: There are few trials in this area of medical care. Only one therapy has been extensively studied, methylprednisolone sodium succinate, which has been shown to improve neurologic outcome up to one year post injury if administered within 8 hours of injury and in a dose regimen of: bolus 30mg/kg administered over 15 minutes with a maintenance infusion of 5.4 mg/kg per hour infused for 23 hours. The initial North American trial was replicated in a Japanese trial but not in the one from France. Data has been obtained from the latter study to permit appropriate meta-analysis of all three trials. This analysis indicates significant recovery in motor function after methylprednisolone therapy. A more recent trial indicates that if methylprednisolone therapy is given for an additional 24 hours (for a total of 48 hours), additional improvement in motor neurologic function and functional status is observed. This is particularly observed if treatment cannot be started until between 3 to 8 hours after injury. The same methylprednisolone therapy has been found effective in whiplash injuries and a modified regimen found to improve recovery after surgery for lumbar disc disease. REVIEWER'S CONCLUSIONS: High dose methylprednisolone steroid therapy is the only pharmacological therapy shown to have efficacy in a Phase Three randomized trial when it can be administered within 8 hours of injury. High dose methylprednisolone has been accepted as standard therapy in many countries. A recent trial indicates additional benefit by extending the maintenance dose from 24 to 48 hours if start of treatment must be delayed to between 3 and 8 hours after injury. There is an urgent need for more randomized trials of pharmacological therapy for acute spinal cord injury. PMID- 10796742 TI - Pool fencing for preventing drowning in children. AB - BACKGROUND: In most industrialized countries, drowning ranks second or third behind motor vehicles and fires as a cause of unintentional injury deaths to children under the age of 15. Death rates from drowning are highest in children less than five years old. Pool fencing is a passive environmental intervention designed to reduce unintended access to swimming pools and thus prevent drowning in the preschool age group. Because of the magnitude of the problem and the potential effectiveness of fencing we decided to evaluate the effect of pool fencing as a drowning prevention strategy for young children. OBJECTIVES: To determine if pool fencing prevents drowning in young children. SEARCH STRATEGY: We used Cochrane Collaboration search strategy of electronic databases, searched reference lists of past reviews and review articles, Cochrane International Register of RCT's, studies from government agencies in the United States and Australia, and contacted colleagues from International Society for Child and Adolescent Injury Prevention, World Injury Network, and CDC funded Injury Control and Research Centers. SELECTION CRITERIA: In order to be selected a study had to be designed to evaluate pool fencing in a defined population and provide relevant and interpretable data which objectively measured the risk of drowning or near drowning or provided rates of these outcomes in fenced and unfenced pools. The completed studies meeting selection criteria employed a case-control design. No randomized controlled studies have been identified. DATA COLLECTION AND ANALYSIS: Three published studies met selection criteria. Data were extracted by two reviewers using standard abstract form. Odds ratios with 95% CI, and incidence rates, were calculated for drowning and near-drowning. Attributable Risk percent (AR%) was calculated to report the reduction in drowning due to pool fencing. MAIN RESULTS: Case control studies which evaluate pool fencing interventions indicate that pool fencing significantly reduces the risk of drowning. Odds ratio for the risk of drowning or near drowning in a fenced pool compared to an unfenced pool is 0.27 95%CI (0.16, 0.47). Isolation fencing (enclosing pool only) is superior to perimeter fencing (enclosing property and pool) because perimeter fencing allows access to the pool area through the house. Odds ratio for the risk of drowning in a pool with isolation fencing compared to a pool with three sided fencing is 0.17 95%CI (0.07, 0.44) REVIEWER'S CONCLUSIONS: Pool fences should have a dynamic and secure gate and isolate (i.e., four-sided fencing) the pool from the house. Legislation should require isolation fencing with secure, self latching gates for all pools, public, semi-public and private. PMID- 10796743 TI - Therapeutic hypothermia for head injury. AB - BACKGROUND: Mild to moderate induced hypothermia has been used in the treatment of head injury for over 50 years, although few randomised controlled trials have been performed. Recent encouraging results from small, single-centre trials and consistent findings of a cerebral protection effect of cooling in laboratory models of global ischaemia has led to a renewed interest in the area. OBJECTIVES: To determine whether the use of mild therapeutic hypothermia in the treatment of moderate and severe head injury improves short-term control of intracranial pressure (ICP) and long-term functional outcome. SEARCH STRATEGY: Electronic searches of the Injuries Group trial registry and EMBASE for any relevant randomised trials, supplemented by hand searching of conference proceedings and reference lists of relevant articles. SELECTION CRITERIA: All randomised controlled trials of mild hypothermia versus control (open or normothermia) in the treatment of patients with any closed head injury requiring hospitalisation. Mild hypothermia was defined as local or systemic cooling to a target temperature of at most 34-35 degrees Celsius for a period of at least 12 hours. Outcome was all-cause mortality and death or severe disability at the end of the scheduled follow-up period. All trials were assessed by two reviewers, and included or excluded on a consensus basis. DATA COLLECTION AND ANALYSIS: Eleven potential trials of therapeutic hypothermia for head injury were found, of which two are ongoing and one is awaiting assessment. The eight remaining trials were included in the systematic review. Data on death, GOS score at final follow-up, complications and ICP were sought and extracted, either from published material or by contact with the investigators. Mantel-Haenzel odds ratios and 95% confidence intervals were calculated for death and death and severe disability for each trial on an intention-to-treat basis. No quantitative synthesis of data on either complications or ICP was attempted. Trials of immediate and deferred hypothermia were analysed separately. MAIN RESULTS: Active immediate hypothermic treatment was associated with a 33% non-significant (p=0.16) reduction in the odds of death at the end of treatment or final follow-up, (OR 0.67, 95% confidence interval 0.38 to 1.17), and a 61% reduction (p=0.004) in the odds of being dead or severely disabled, (OR 0.39, 95% confidence interval 0.20 to 0.74). Similar effect sizes were found for delayed hypothermia. These results are, however, based on a few small trials each of less than 100 patients. A multi centre trials of hypothermia versus control in 392 patients will be reporting results in 1999, providing substantially more evidence than is currently available. REVIEWER'S CONCLUSIONS: Although this review would suggest a strong positive effect of therapeutic hypothermia, the results are based on several small trials carried out in single, specialist centres. The results of a large multi-centre trial are expected in 1999 and will more than treble the available evidence. Until these results have been released, it would be inappropriate to make any short-term recommendations for clinical practice or research. PMID- 10796744 TI - Mannitol for acute traumatic brain injury. AB - BACKGROUND: Mannitol is sometimes dramatically effective in reversing acute brain swelling, but its effectiveness in the on-going management of severe head injury remains open to question. There is evidence that in prolonged dosage mannitol may pass from the blood into the brain, where it might cause reverse osmotic shifts that increase intracranial pressure. OBJECTIVES: To assess the effects of different mannitol therapy regimens, of mannitol compared to other intracranial pressure (ICP) lowering agents, and to quantify the effectiveness of mannitol administration given at other stages following acute traumatic brain injury. SEARCH STRATEGY: The review drew on the search strategy for the Injuries Group as a whole. We checked reference lists of trials and review articles, and contacted authors of trials. SELECTION CRITERIA: Randomised trials of mannitol, in patients with acute traumatic brain injury of any severity. The comparison group could be placebo-controlled, no drug, different dose, or different drug. Trials where the intervention was started more than eight weeks after injury, and cross-over trials were excluded. DATA COLLECTION AND ANALYSIS: The reviewers independently rated quality of allocation concealment and extracted the data. Relative risks (RR) and 95% confidence intervals (CI) were calculated for each trial on an intention to treat basis. MAIN RESULTS: Overall there were few eligible trials. There were no trials comparing different doses, or type of administration. One trial compared ICP-directed therapy to 'standard care' (RR for death= 0.83; 95% CI 0.47;1.46). One trial compared mannitol to pentobarbital (RR for death = 0.85; 95% CI 0. 52;1.38). No trials compared mannitol to other ICP lowering agents. One trial tested the effectiveness of pre-hospital administration of mannitol against placebo (RR for death=1.59; 95% CI 0.44;5.79). REVIEWER'S CONCLUSIONS: There are insufficient data to recommend one form of mannitol infusion over another. Mannitol therapy for raised ICP may have a beneficial effect on mortality when compared to pentobarbital treatment. ICP-directed treatment shows a small beneficial effect compared to treatment directed by neurological signs and physiological indicators. There are insufficient data on the effectiveness of pre hospital administration of mannitol to preclude either a harmful or a beneficial effect on mortality. PMID- 10796745 TI - Community screening for visual impairment in the elderly. AB - BACKGROUND: While the aims of multicomponent screening of older people are broad, any benefit arising from the inclusion of a vision component in the assessment will necessarily be dependent on improved vision. OBJECTIVES: The objective of this review is to assess the effects on vision of mass screening of older people for visual impairment. SEARCH STRATEGY: We searched the Cochrane Eye and Vision Group specialised register, the Cochrane Controlled Trials Register - Central, MEDLINE, EMBASE, SciSearch and reference lists of relevant trial reports and review articles. We contacted investigators to identify additional published and unpublished trials. The most recent searches were conducted in April 1998. SELECTION CRITERIA: We included randomised trials of visual or multicomponent screening for vision impairment in people aged 65 or over in a community setting. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Visual outcome data were available for 3494 people in five trials of multicomponent assessment. Length of follow up ranged from two to four years. All the trials used self-reported measures for visual impairment, both as screening tools and as outcome measures. In four of the trials people reporting visual problems were referred to either the eye services or to a physician. In one trial people reporting visual problems received information about resources in the community designed to assist those with poor vision. The proportions of participants in the intervention and control groups who reported visual problems at the time of outcome assessment were 0.26 and 0.23 respectively (relative risk for visual impairment 1.03, 95% confidence interval 0.92 to 1.15). REVIEWER'S CONCLUSIONS: There is no evidence that community-based screening of asymptomatic older people results in improvements in vision. PMID- 10796746 TI - Laparoscopic "drilling" by diathermy or laser for ovulation induction in anovulatory polycystic ovary syndrome. AB - BACKGROUND: Problems in inducing ovulation in women with polycystic ovary syndrome (PCOS) and anovulation (failure to ovulate) are well recognised. Surgical ovarian wedge resection was the first established treatment for anovulatory PCOS patients but was largely abandoned of the risk of post-surgical adhesion formation. It was replaced by medical ovulation induction with clomiphene and gonadotrophins. However patients with PCOS treated with gonadotrophins often have a polyfollicular response and are exposed to the risks of ovarian hyperstimulation syndrome (OHSS) and multiple pregnancy. Although effective, it is an expensive, stressful and time consuming form of treatment requiring intensive monitoring. A new surgical therapy, laparoscopic ovarian "drilling", may avoid or reduce the need, or facilitate the use, of gonadotrophins for inducing ovulation. The procedure can be done on an outpatient basis with less trauma and fewer postoperative adhesions. It has been claimed in many uncontrolled observational studies that it is followed, at least temporarily, by a high rate of spontaneous postoperative ovulation and conception, or that subsequent medical ovulation induction becomes easier. OBJECTIVES: To determine the effectiveness of laparoscopic ovarian drilling for ovulation induction in subfertile women with anovulation (failure to ovulate) and polycystic ovarian syndrome (PCOS). SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility Group was used for the identification of randomised controlled trials (RCTS). A computerised MEDLINE search was used to identify non randomised controlled trials. SELECTION CRITERIA: Trials were eligible for inclusion if treatment consisted of laparoscopic ovarian drilling in order to induce ovulation in subfertile women with PCOS and compared with a concurrent control group. DATA COLLECTION AND ANALYSIS: Fourteen trials were identified; eight were included in the review of which seven were randomised. All trials were assessed for quality criteria. The main studied outcomes were ovulation and pregnancy rates. Miscarriage rate, multiple pregnancy rate, and incidence of overstimulation and ovarian hyperstimulation syndrome rate were secondary outcomes. MAIN RESULTS: With the exception of multiple pregnancy rates no differences were demonstrated for any of the interventions studied but the numbers of patients who have been randomised to controlled studies at this time is insufficient to conclude that laparoscopic ovarian drilling is more effective than gonadotrophin therapy for other outcomes. REVIEWER'S CONCLUSIONS: The value of laparoscopic ovarian drilling as a primary treatment for subfertile patients with anovulation (failure to ovulate) and polycystic ovarian syndrome (PCOS) is undetermined. There is insufficient evidence to determine a difference in ovulation or pregnancy rates when compared to gonadotrophin therapy as a secondary treatment for clomiphene resistant women. Multiple pregnancy rates are reduced in those women who conceive following laparoscopic drilling. None of the studied modalities of drilling technique had any obvious advantages. PMID- 10796747 TI - Pre-operative endometrial thinning agents before hysteroscopic surgery for heavy menstrual bleeding. AB - BACKGROUND: Menorrhagia is one of the most common reasons for pre-menopausal women to be referred to a gynaecologist. Although medical therapy is generally the first approach, many will eventually require or request a hysterectomy. Hysterectomy is associated with a significant in-patient hospital stay and a period of convalescence that makes it an unattractive and unnecessarily invasive option for many women. Hysteroscopic endometrial ablation or resection offers a day-case surgical alternative to hysterectomy for these women. It is also a cheaper procedure than hysterectomy. Complete endometrial removal or destruction is one of the most important determinants of treatment success. Therefore surgery will be most effective if undertaken when endometrial thickness is less than 4mm, in the immediate post-menstrual phase, however there are often difficulties in reliably arranging surgery for this time. The other option is the use of hormonal agents which induce endometrial thinning or atrophy prior to surgery. The most commonly evaluated agents have been goserelin (a GnRH analogue) and danazol. Progestogens and other GnRH analogues have also been studied although less data are available. It has been suggested that the use of these agents, particularly GnRH analogues, will reduce operating time, improve the intra-uterine operating environment, and reduce distension medium absorption (this is the fluid used to distend the uterine cavity during surgery). OBJECTIVES: To investigate the effectiveness of gonadotrophin-releasing hormone (GnRH) analogues, danazol, and progestogens, when used for endometrial thinning prior to hysteroscopic surgery for menorrhagia, in improving the intra-uterine operating environment and treatment outcome after surgery. SEARCH STRATEGY: The Menstrual Disorders and Subfertility Group search strategy (see Review Group details) was used to identify randomised trials that had compared the use of these drugs with either each other, or placebo, or no pre-operative treatment. SELECTION CRITERIA: Trials were included if they compared the effects of these agents with each other, or with placebo or no treatment on relevant intra-operative and post-operative treatment outcomes. Only randomised studies were included in this review. DATA COLLECTION AND ANALYSIS: Eight studies met the inclusion criteria for this review. Four studies compared goserelin (a GnRH analogue) with no treatment or placebo. Three studies compared goserelin with danazol. One study compared progestogens, danazol and triptorelin (a GnRH analogue) with no treatment. Data was extracted independently by two reviewers. A third reviewer checked data extraction for accuracy and wrote to authors where relevant data was missing or unclear. Intra-operative parameters included endometrial thickness, duration of surgery, ease of surgery, distension medium absorption and complication rate. Post-operative outcomes compared were the proportion of women with amenorrhoea, post-operative menstrual loss and dysmenorrhoea, and the need for further surgery. Data on side-effects were also recorded. MAIN RESULTS: When compared with no treatment GnRH analogues are associated with a shorter duration of surgery, greater ease of surgery and a higher rate of post-operative amenorrhoea. Post-operative dysmenorrhoea also appears to be reduced. The use of GnRH analogues has no effect on intra-operative complication rates and patient satisfaction with this surgery is high irrespective of the use of any pre operative endometrial thinning agent. GnRH analogues produce more consistent endometrial atrophy than danazol. For other intra-operative and post-operative outcomes any differences are minimal. Both GnRH analogues and danazol produce side-effects in a significant proportion of women, though few studies have reported these in detail. Little randomised data is available to assess the effectiveness of progestogens as endometrial thinning agents and the effect of any thinning agent PMID- 10796748 TI - Deoxyribonuclease for cystic fibrosis. AB - BACKGROUND: Recombinant human deoxyribonuclease is currently used to treat pulmonary disease (the major cause of morbidity and mortality) in cystic fibrosis. OBJECTIVES: To determine whether the use of recombinant human deoxyribonuclease in cystic fibrosis is associated with improved mortality and morbidity as compared to placebo and to identify any adverse events associated with its use. To compare the efficacy of recombinant human deoxyribonuclease with other mucolytics. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, hand searching relevant journals and abstracts from conferences. The company producing recombinant human deoxyribonuclease was also contacted. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised and quasi-randomised trials where recombinant human deoxyribonuclease was compared to either placebo, standard therapy or another mucolytic for any duration, dose regimen and age of patient with cystic fibrosis of any disease severity. DATA COLLECTION AND ANALYSIS: Trials were independently assessed for inclusion criteria, methodological quality and data extraction by the two reviewers. Comparisons were between recombinant human deoxyribonuclease and placebo and recombinant human deoxyribonuclease and other mucolytics. The following outcomes were recorded: Mean % change from baseline in forced vital capacity (FVC), forced expiratory voloume at one second (FEV1) and weight, mean number of respiratory tract exacerbations, days intravenous and oral antibiotics used, mean number of days as inpatient, number of deaths, adverse events and the cost of therapy. MAIN RESULTS: Seven primary clinical trials were identified, totalling 1710 patients. Two further studies examined the health care cost of patients from one of the clinical trials. No eligible studies compared recombinant human deoxyribonuclease to another mucolytic. Five trials presented outcomes at up to one month, one at three months and one at six months. No reduction in mortality for treated patients was identified (Relative Risk (RR) at six month 1.01, 95%Confidence Interval (CI) 0.09, 11.11). Lung function improved to a greater extent in the treated groups (at six months Weighted Mean Difference (WMD) FEV1 5.7, 95%CI 4.18, 7.23, at three months 7.3, 95%CI 4.04, 10.65). Pooled data from the five trials of up to one month gave WMD 9.2 95%CI 0.93, 17. 6 although there was significant heterogeneity). Recombinant human deoxyribonuclease was well tolerated with no excess of serious adverse events (RR haemoptysis 0.89, 95%CI 0.54, 1.45, pneumothorax 0.97 95%CI 0.19, 4.96). Voice alteration was, however, reported more frequently in the treated groups (RR 2.33 95%CI 1.38, 3.93). No study analysed our pre-defined outcome measure for respiratory exacerbations and insufficient data was available to analyse differences in antibiotic treatment, inpatient stay and quality of life. REVIEWER'S CONCLUSIONS: Studies are of insufficient duration to identify a reduction in mortality or number of respiratory exacerbations. Further trials are required to answer these important questions. Recombinant human deoxyribonuclease therapy is associated with an improvement in lung function after six months treatment, but it is not possible to assess whether this effect on lung function is sustained in the long-term. No studies were identified that compared recombinant human deoxyribonuclease to another mucolytic. PMID- 10796749 TI - A comparison of drugs versus placebo for the treatment of dysthymia. AB - OBJECTIVES: Dysthymia is a depressive disorder of chronic nature but of less severity than major depression, which depressive symptoms are more or less continuous for at least two years. The aim of this review was to conduct a systematic review of all RCTs comparing drugs and placebo for dysthymia. SEARCH STRATEGY: Electronic searches of Cochrane Library, EMBASE, MEDLINE, PsycLIT, Biological Abstracts and LILACS; reference searching; personal communication; conference abstracts; unpublished trials from the pharmaceutical industry; book chapters on the treatment of depression. SELECTION CRITERIA: The inclusion criteria for all randomised controlled trials were that they should focus on the use of drugs versus placebo for dysthymic patients. Exclusion criteria were: non randomised, mixed major depression/ dysthymia (trials not providing separate data) and depression secondary to other disorders (e.g. substance abuse). DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently. In order to achieve an intention-to-treat analysis, when trials failed to report it was assumed that people who died or dropped out had no improvement. Authors of relevant trials were contacted for additional and missing data. Absence of treatment response as defined by authors was the main measure of outcome used. Relative Risks (RR) and 95% confidence intervals (CI) of dichotomous data were calculated with the Random Effects Model. Where possible, number needed to treat (NNT) and number needed to harm (NNH) were estimated, taking the reciprocal of the absolute risk reduction. MAIN RESULTS: Currently the review includes 15 trials. Similar results were obtained in terms of efficacy for different groups of drugs, such as tricyclic (TCA), selective serotonin reuptake inhibitors (SSRI), monoamine oxidase inhibitors (MAOI) and other drugs (sulpiride, amineptine, and ritanserin). The pooled RR for absence of treatment response was 0. 68 (95% CI 0.59-0.78) for TCA and the NNT was 4.3 (95% CI 3.2-6.5). SSRIs showed similar RR for this outcome: 0.64 (95% CI 0.55-0.74), the NNT being 4.7 (95% CI 3.5-6.9). Concerning MAOIs, the RR was 0. 59 (95% CI 0.48-0.71) and the NNT was 2.9 (95% CI 2.2-4.3). Other drugs (amisulpride, amineptine and ritanserin) showed similar results in terms of absence of treatment response. Using more stringent criteria for improvement - full remission - the results were unchanged. Patients treated on TCA were more likely to report adverse events, compared with placebo. REVIEWER'S CONCLUSIONS: Drugs are effective in the treatment of dysthymia with no differences between and within class of drugs. Tricyclic antidepressants are more likely to cause adverse events and dropouts. As dysthymia is a chronic condition, there remains little information on quality of life and medium or long-term outcome. PMID- 10796750 TI - Pharmacological interventions for spasticity following spinal cord injury. AB - BACKGROUND: Spasticity is a major health problem for patients with a spinal cord injury (SCI) that limits patients' mobility and affects independence in activities of daily living and work. Spasticity may also cause pain, loss of range of motion, contractures, sleep disorders and impair ambulation in patients with an incomplete lesion. The effectiveness of available drugs is still uncertain and they may cause adverse effects. Assessing what works in this area is complicated by the lack of valid and reliable measurement tools. The aim of this systematic review is to critically appraise and summarise existing information of the effectiveness of available treatments and to identify areas where further research is needed. OBJECTIVES: To assess the effectiveness and safety of Baclofen, Dantrolene, Tizanidine and any other drugs for the treatment of long term spasticity in SCI patients as well as the effectiveness and safety of different routes of administration of Baclofen. SEARCH STRATEGY: We searched the Injuries Group specialised register, the Cochrane Controlled Trials Register, MEDLINE, EMBASE and CINHALH up to 1998. Drug companies and experts active in the area were also contacted. SELECTION CRITERIA: All parallel and crossover RCTs including spinal cord injury patients complaining of "severe spasticity". Studies where less than 50% of patients had a spinal cord injury were excluded. DATA COLLECTION AND ANALYSIS: Methodological quality of studies (allocation concealment, blinding, patients characteristics, inclusion and exclusion criteria; interventions; outcomes; lost to follow up) was independently assessed by two investigators. The heterogeneity among studies did not allow quantitative combination of results. MAIN RESULTS: Nine out of 53 studies met the inclusion criteria. Study design was: 8 cross over, 1 parallel-group trial. Two studies (14 SCI patients), showed a significant effect of intrathecal baclofen in reducing spasticity (Ashworth Score and ADL performances), compared to placebo, without any side effect. The study comparing tizanidine to placebo (118 SCI patients) showed a significant effect of tizanidine in improving Ashworth Score but not in ADL performances. Tizanidine group reported significant rates of adverse effects (drowsiness, xerostomia). For the other drugs (Gabapentine, Clonidine, Diazepam, Amytal and oral Baclofen ) the results do not provide evidence for a clinical significant effectiveness. REVIEWER'S CONCLUSIONS: There is insufficient evidence to assist clinicians in a rational approach to antispastic treatment for SCI. Further research is urgently needed to improve the scientific basis of patient care. PMID- 10796751 TI - Post-operative 5-fluorouracil for glaucoma surgery. AB - BACKGROUND: Trabeculectomy is performed as a treatment for many types of glaucoma in an attempt to lower the intra-ocular pressure. The surgery involves creating a channel through the sclera, through which intra-ocular fluid can leave the eye. If scar tissue blocks the exit of the surgically created channel intra-ocular pressure rises and the operation fails. Antimetabolites are used to inhibit wound healing to prevent the conjunctiva scarring down on to the sclera. The principal antimetabolites used are 5-Fluorouracil and Mitomycin C. Both may be applied on a sponge between the conjunctiva and sclera at the beginning of surgery. 5 Fluorouracil may also be given as one or more injections under the conjunctiva after the surgery. OBJECTIVES: The objective of this review is to assess the effects of post-operative injections of 5-Fluorouracil in eyes of people undergoing surgery for glaucoma. SEARCH STRATEGY: We searched the Cochrane Eyes and Vision Group specialised register, The Cochrane Controlled Trials Register (CENTRAL), MEDLINE and EMBASE. We searched the reference lists of relevant articles for additional trials, and we used the Science Citation Index to search for articles that cited the included studies. We contacted investigators and experts for details of additional relevant trials. SELECTION CRITERIA: We included randomised trials of post-operative 5-Fluorouracil injections compared to placebo injections or no injections in trabeculectomy for glaucoma. DATA COLLECTION AND ANALYSIS: We independently assessed trial quality and extracted data. We contacted trial investigators for missing information. Data were summarised using relative risk, Peto odds ratio and weighted mean difference as appropriate. MAIN RESULTS: Eight trials, which randomised a total of 536 participants, are included in the review. As far as can be determined from the trial reports, the methodological quality of the trials was not high. 5 Fluorouracil when delivered by post-operative injection appears to be effective in reducing the likelihood of surgical failure of trabeculectomy both in eyes at high risk of failure and those undergoing surgery for the first time. Complications are more common after 5-Fluorouracil injections, especially early and late wound leaks and temporary damage to the ocular surface. Whilst no evidence was found of an increased risk of serious sight threatening complications, people undergoing initial trabeculectomy had a substantially increased risk of ocular hypotony. None of the trials reported on the participants' perspective of care. REVIEWER'S CONCLUSIONS: This treatment is now rarely used on the basis of a planned series of post-operative injections.This presumably reflects an aspect of the treatment that is unacceptable to both patients and doctors. None of the trials reported on the participants' perspective of care which constitutes a serious omission for an invasive treatment such as this. Increasingly, injections are used on an ad hoc basis. The effectiveness of this strategy is the subject of a future systematic review. PMID- 10796752 TI - Anticonvulsant drugs for acute and chronic pain. AB - BACKGROUND: Anticonvulsant drugs have been used in the management of pain since the 1960s. The clinical impression is that they are useful for neuropathic pain, especially when the pain is lancinating or burning. OBJECTIVES: To evaluate the analgesic effectiveness of anticonvulsant drugs compared to either placebo or other drugs in order to provide evidence-based recommendations for pain management in clinical practice and to identify a clinical research agenda. Adverse effects are also considered. SEARCH STRATEGY: Randomised trials of anticonvulsants in acute, chronic or cancer pain were identified by Medline (Silver Platter 3.0, 3.1 and 3.11) from 1966 to February 1994. In addition, 40 medical journals were hand searched (published between 1950 and 1990). Additional reports were identified from the reference list of the retrieved papers, and contacting investigators. Date of the most recent searches: 1994. SELECTION CRITERIA: Randomised trials reporting the analgesic effects of anticonvulsant drugs in patients, with pain assessment as either the primary or a secondary outcome. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers, and trials were quality scored. Numbers-needed-to-treat (NNTs) were calculated from dichotomous data for effectiveness, adverse effects and drug related study withdrawal, for individual studies and for pooled data. MAIN RESULTS: Twenty trials of four anticonvulsants were considered eligible (746 patients). The only placebo-controlled study in acute pain found no analgesic effect of sodium valproate. Three placebo-controlled studies of carbamazepine in trigeminal neuralgia had a combined NNT for effectiveness of 2.6, for adverse effects 3.4, and for severe effects (withdrawal from study) 24. Three placebo controlled studies of diabetic neuropathy had a combined NNT for effectiveness of 3, for adverse effects 2.5, and for severe effects 20. Three placebo-controlled studies of migraine prophylaxis had a combined NNT for effectiveness of 2.4, for adverse effects 2.4 and for severe effects 39. Phenytoin had no effect in irritable bowel syndrome, and carbamazepine little effect in post-stroke pain. Clonazepam was effective in one study of temporomandibular joint dysfunction. No study compared one anticonvulsant with another. Anticonvulsants fared poorly against other treatments. REVIEWER'S CONCLUSIONS: Although anticonvulsants are used widely in chronic pain surprisingly few trials show analgesic effectiveness. No trial compared different anticonvulsants. There is no evidence that anticonvulsants are effective for acute pain. In chronic pain syndromes other than trigeminal neuralgia anticonvulsants should be withheld until other interventions have been tried. PMID- 10796753 TI - Enteral tube feeding for cystic fibrosis. AB - BACKGROUND: Enteral tube feeding is routinely used in many cystic fibrosis centres when weight for height percentage is less than 85%, when there has been weight loss for greater than a two month period or when there has been no weight gain for two to three months (under five years old) or for six months (over five years old). OBJECTIVES: To examine the evidence that in patients with cystic fibrosis supplemental enteral tube feeding improves nutritional status, respiratory function, and quality of life without significant adverse effects. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialised register and contacted the companies which market enteral feeds and reviewed their databases. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised controlled trials comparing supplemental enteral tube feeding for one month or longer with no specific intervention in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: There are no trials included in this review. MAIN RESULTS: There are no trials included in this review. REVIEWER'S CONCLUSIONS: Supplemental enteral tube feeding is widely used throughout the world to improve nutritional status in patients with cystic fibrosis. The methods mostly used, nasogastric or gastrostomy feeding, are invasive, expensive, and may have a negative effect on self esteem and body image. Reported use of enteral tube feeding suggests that it results in nutritional and respiratory improvement and it is disappointing that their efficacy has not been fully assessed by randomised controlled trials. With the more frequent recommendations to use enteral tube feeding as an early rather than a late intervention, this systematic review identifies the need for a multi-centre randomised controlled trial assessing both efficacy and possible adverse effects of enteral tube feeding in cystic fibrosis. PMID- 10796754 TI - Neoadjuvant and adjuvant therapy for operable hepatocellular carcinoma. AB - OBJECTIVES: To determine the efficacy and adverse effects of different neoadjuvant and adjuvant therapies compared to surgery alone or surgery and placebo/supportive therapy when given to improve relapse and survival rates for operable hepatocellular carcinoma. SEARCH STRATEGY: Electronic databases, conference proceedings, bibliographies of identified publications. SELECTION CRITERIA: All truly randomised and quasi-randomised clinical trials that compared hepatocellular carcinoma patients who were given and not given neoadjuvant/adjuvant therapy as a supplement to curative liver resection. DATA COLLECTION AND ANALYSIS: Study data was extracted independently by two reviewers and discrepancies were resolved by consensus. A total of eight randomised controlled clinical trials were identified, totaling 548 randomised patients. Seven of the eight trials reported survival and disease-free survival curves and the results of hypothesis testing (log-rank test). The remaining trial reported only the mean survival times. None reported the hazard ratio and only one did a sample size calculation. The survival and disease-free survival curves were compared using their one, two and three-year survival rates, median survival times and the result of the hypothesis tests. MAIN RESULTS: The size of the randomised clinical trials ranged from 40 to 115 subjects. Both preoperative (neoadjuvant) and postoperative (adjuvant), systemic and locoregional (+/- embolization), chemo- and immunotherapy interventions were tested. None were comparable in terms of both treatment regimen and participants selected, so no pooling was done. Only one regimen using preoperative transcatheter arterial chemoembolization with doxorubicin was approximately duplicated. Seven of the eight trials reported no survival benefit from adjuvant therapy. Only one trial reported a statistically significant difference for survival and disease-free survival for the treatment arm, but the results of both its arms were very poor when compared to other studies. Two of the trials that did not report any absolute survival advantage reported statistically significant differences in disease-free survival. Five of the eight trials did not perform intention-to treat analysis. The highest toxicity rate was in a trial using oral 1 hexylcarbamoyl 5-fluorouracil which resulted in 12 out of 38 subjects stopping because of adverse events. REVIEWER'S CONCLUSIONS: There is no evidence for efficacy of any of the adjuvant protocols reviewed. In order to detect a realistic treatment advantage, larger trials will have to be conducted. PMID- 10796755 TI - Bromocriptine for levodopa-induced motor complications in Parkinson's disease. AB - OBJECTIVES: To assess the efficacy and safety of adjunct bromocriptine (BR) therapy compared to placebo in the treatment of Parkinson's disease (PD) patients with motor complications. SEARCH STRATEGY: Sources including the Cochrane Library, a MEDLINE search-strategy, reference lists of the reviews found by the MEDLINE search-strategy, Sandoz (producer of BR), symposia reports, PD handbooks, SCISEARCH, contacts with colleagues who had co-ordinated trials on BR and reference lists of all included studies were used to identify randomized controlled trials (RCTs) of interest. SELECTION CRITERIA: Randomized trials were eligible for inclusion if they evaluated the efficacy of BR as adjunctive to LD therapy compared to placebo in PD patients with motor complications. Outcome measures that were evaluated, included occurrence and severity of motor complications, scores on impairment and disability, and the occurrence of side effects. DATA COLLECTION AND ANALYSIS: Three reviewers independently reviewed the quality of identified trials. To determine the feasibility of a quantitative systematic review each eligible study was evaluated concerning the methodological quality. MAIN RESULTS: This review identified important shortcomings regarding the methodological quality of eight trials. All studies failed to describe adequately their randomization procedure. Consultation with the trialists revealed that three trials adequately randomized their patients. Contrary to the information of the published report, one placebo-controlled trial appeared to be carried out as an open study and was therefore excluded. The remaining seven trials were reported to be carried out according to a double-blind design, although one was unblinded after five weeks. There was a conspicuous variability in the duration of trials: four to forty weeks (mean 14 weeks). None of the included trials was performed according to the intention-to-treat principle. With regard to the inclusion criteria, it frequently remained unclear if PD patients actually suffered from motor complications. Prominent differences between studies regarding the baseline characteristics and the rate by which BR was introduced during the titration phase were found. Major differences between studies emerged concerning the applied outcomes. The various methods used to evaluate the occurrence and/or severity of motor complications lacked a sound clinimetric basis. A great diversity of scales to evaluate impairment and disability was applied. None of the included trials reported whether scores on impairment and disability level referred to the "on"- or "off"-phase. REVIEWER'S CONCLUSIONS: This review highlights major methodological problems and sources of heterogeneity that not only hamper the comparability of trials but also preclude a conclusion on the efficacy of BR in the adjunct treatment of PD patients with motor complications. PMID- 10796756 TI - Human albumin solution for resuscitation and volume expansion in critically ill patients. The Albumin Reviewers. AB - BACKGROUND: Human albumin solutions are used in a range of medical and surgical problems. Licensed indications are the emergency treatment of shock and other conditions where restoration of blood volume is urgent, burns, and hypoproteinaemia. Human albumin solutions are more expensive than other colloids and crystalloids. OBJECTIVES: To quantify the effect on mortality of human albumin and plasma protein fraction (PPF) administration in the management of critically ill patients. SEARCH STRATEGY: We searched the Cochrane Injuries Group trials register, Cochrane Controlled Trials Register, Medline, Embase and BIDS Index to Scientific and Technical Proceedings. Reference lists of trials and review articles were checked, and authors of identified trials were contacted. The search was last updated in November 1999. SELECTION CRITERIA: Randomised controlled trials comparing albumin/PPF with no albumin/PPF, or with a crystalloid solution, in critically ill patients with hypovolaemia, burns or hypoalbuminaemia. DATA COLLECTION AND ANALYSIS: We collected data on the participants, albumin solution used, mortality at the end of follow up, and quality of allocation concealment. Analysis was stratified according to patient type. We assessed publication bias using the regression test for funnel plot asymmetry. MAIN RESULTS: We found 30 trials meeting the inclusion criteria and reporting death as an outcome. There were 156 deaths among 1419 trial participants. For each patient category the risk of death in the albumin treated group was higher than in the comparison group. For hypovolaemia the relative risk of death following albumin administration was 1.46 (95% confidence interval 0.97 to 2.22), for burns the relative risk was 2.40 (1.11 to 5.19), and for hypoalbuminaemia the relative risk was 1.69 (1.07 to 2.67). The pooled relative risk of death with albumin administration was 1.68 (1.26 to 2.23). Overall, the risk of death in patients receiving albumin was 14% compared to 8% in the control groups, an increase in the risk of death of 6% (3% to 9%). These data suggest that for every 17 critically ill patients treated with albumin there is one additional death. REVIEWER'S CONCLUSIONS: There is no evidence that albumin administration reduces the risk of death in critically ill patients with hypovolaemia, burns or hypoalbuminaemia, and a strong suggestion that it may increase the risk of death. These data suggest that the use of human albumin in critically ill patients should be urgently reviewed and that it should not be used outside the context of a rigorously conducted randomised controlled trial. PMID- 10796757 TI - Antibiotics for acute bacterial conjunctivitis. AB - BACKGROUND: There are concerns regarding whether antibiotic therapy confers significant benefit in the treatment of acute bacterial conjunctivitis. OBJECTIVES: The aim of this review is to assess the benefit and harm of antibiotic therapy in the management of acute bacterial conjunctivitis. SEARCH STRATEGY: We searched the Cochrane Eyes and Vision Group specialised register, the Cochrane Controlled Trials Register - Central, MEDLINE and the reference lists of identified trial reports. We used the Science Citation Index to look for articles that cited the relevant studies, and we contacted investigators and pharmaceutical companies for information about additional trials. The most recent searches were carried out in September 1998. SELECTION CRITERIA: We included double masked randomised controlled trials in which any form of antibiotic treatment had been compared with placebo in the management of acute bacterial conjunctivitis. This included topical, systemic and combination (for example, antibiotics and steroids) antibiotic usage. DATA COLLECTION AND ANALYSIS: One reviewer extracted data and the accuracy was checked by a second reviewer. Relative risks were summarised. We tested for heterogeneity between studies. MAIN RESULTS: Six published trials were identified of which three fulfilled the eligibility criteria for inclusion in this review. One trial was single masked and therefore excluded. A second report, when translated, was found to have no placebo group and was therefore excluded. One trial is currently 'awaiting assessment'. This has been published in abstract form and has yet to be fully reported. All the trials thus far identified appear to have been conducted on a selected specialist care patient population. The trials were heterogeneous in terms of their inclusion and exclusion criteria, the nature of the intervention, and the outcome measures assessed. Meta-analysis indicates that acute bacterial conjunctivitis is frequently a self-limiting condition, as clinical remission (cure or significant improvement) occurred by days two to five in 64% (95% confidence interval (CI) 57% to 71%) of those treated with placebo. Treatment with antibiotics was, however, associated with significantly better rates of clinical remission (days two to five: relative risk (RR) 1.31 95% CI 1.11 to 1.55, NNT=5) with a suggestion that this benefit was maintained for late clinical remission (days six to 10: RR 1.27 95% CI 1.00 to 1.61, NNT=5). Antibiotic treatment was associated with rates of microbiological remission (pathogen eradication or reduction). No serious outcomes were reported in either the active or placebo arms of these trials, indicating that important sight-threatening complications are an infrequent occurrence. REVIEWER'S CONCLUSIONS: Acute bacterial conjunctivitis is frequently a self-limiting condition but the use of antibiotics is associated with significantly improved rates of early clinical remission and early and late microbiological remission. Since trials to-date have been conducted in selected specialist care patient populations these results may not necessarily be generalisable to a primary care based population. A trial based in primary care designed to assess the cost-effectiveness of commonly prescribed antibiotic(s) versus placebo in acute bacterial conjunctivitis is warranted. PMID- 10796758 TI - Interventions for guttate psoriasis. AB - BACKGROUND: Guttate psoriasis is a distinctive acute form of psoriasis which characteristically occurs in children and young adults. Very little specific evidence-based guidance is available in standard texts to help make rational decisions about treatment options. OBJECTIVES: To assess the effectiveness of treatments for guttate psoriasis. SEARCH STRATEGY: We searched the Cochrane Clinical Trials Register (Cochrane Library, Issue 3, 1999), Medline (1966- September 1999), Embase (1988-September 1999), Salford Database of Psoriasis Trials (to November 1999) and European Dermato-Epidemiology Network (EDEN) Psoriasis Trials Database (to November 1999) for terms GUTTATE and PSORIASIS. We also searched 100 unselected RCTs of psoriasis therapy and all 112 RCTs of phototherapy for psoriasis in the Salford Database of Psoriasis Trials for separate stratification for guttate psoriasis. SELECTION CRITERIA: Randomised trials in which patients with acute guttate psoriasis were randomised to different treatments, except those trials examining antistreptococcal interventions which are addressed in a separate Cochrane review. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality. MAIN RESULTS: No published report could be found to support or to challenge current commonly used methods of management. Only one trial which met the selection criteria was identified. In this small study of 21 hospitalised patients with guttate psoriasis, intravenous infusion of an n-3 fatty acid rich lipid emulsion was compared with placebo emulsion containing n-6 fatty acids. The n-3 preparation appeared to be of some benefit for patients with guttate psoriasis. REVIEWER'S CONCLUSIONS: There is currently no firm evidence on which to base treatment of acute guttate psoriasis. Studies comparing standard treatment modalities, including phototherapy and topical regimens, are required to enable informed decisions on treatment choices to be made. PMID- 10796759 TI - Systemic treatments for metastatic cutaneous melanoma. AB - BACKGROUND: Systemic therapies for metastatic cutaneous melanoma, the most aggressive of all skin cancers, remain disappointing. Few lasting remissions are achieved and the therapeutic aim remains one of palliation. Many agents are used alone or in combination with varying degrees of toxicity and cost. It is unclear whether evidence exists to support these complex regimens over best supportive care / placebo. OBJECTIVES: To review the benefits from the use of systemic therapies in metastatic cutaneous melanoma compared to best supportive care/placebo, and to establish whether a 'standard' therapy exists which is superior to other treatments. SEARCH STRATEGY: Randomised controlled trials were identified from the MEDLINE, EMBASE and CCTR/CENTRAL databases. References, conference proceedings, and Science Citation Index/Scisearch were also used to locate trials. Cancer registries and trialists were also contacted. SELECTION CRITERIA: Randomised controlled trials of adults with histologically proven metastatic cutaneous melanoma in which systemic anti-cancer therapy was compared with placebo or supportive care. DATA COLLECTION AND ANALYSIS: Study selection was performed by two independent reviewers. Data extraction forms were used for studies which appeared to meet the selection criteria and, where appropriate, full text articles were retrieved and reviewed independently. MAIN RESULTS: No randomised controlled trials were found comparing a systemic therapy with placebo or best supportive care in metastatic cutaneous melanoma. REVIEWER'S CONCLUSIONS: There is no evidence from randomised controlled clinical trials to show superiority of systemic therapy over best supportive care / placebo in the treatment of malignant cutaneous melanoma. Given that patients with metastatic melanoma frequently receive systemic therapy, it is our pragmatic view that a future systematic review could compare any systemic treatment, or combination of treatments, to single agent dacarbazine. PMID- 10796760 TI - Screening for colorectal cancer using the faecal occult blood test, hemoccult. AB - BACKGROUND: Colorectal cancer is a leading cause of illness and death in the Western world. In Australia, the United Kingdom and the United States, it is the second commonest cancer for women after breast cancer (age-standardised incidence 22-33 per 100,000), and men after prostate or lung cancer (age-standardised incidence 31-47 per 100,000) (Jeffs et al, 1996; Parkin et al, 1992). Just under half of all persons affected will die from their disease (Jeffs et al, 1996; Parkin et al, 1992) The human and financial costs of this disease have prompted considerable research efforts to evaluate the ability of screening tests to detect the cancer at an early curable stage. Tests which have been considered for screening include faecal occult blood tests, sigmoidoscopy and colonoscopy. OBJECTIVES: To determine whether screening for colorectal cancer using the faecal occult blood test, Hemoccult reduces colorectal cancer mortality and to consider the benefits and harms of screening. SEARCH STRATEGY: Published and unpublished data for this review were identified by: * retrieving studies included in a systematic review conducted by some of the authors in 1995, * searches of MEDLINE, Current Contents and the Cochrane Controlled Trials Register, * writing to trial lists. SELECTION CRITERIA: All controlled trials of screening for colorectal cancer using Hemoccult were eligible for inclusion in the review. DATA COLLECTION AND ANALYSIS: Data from the trials were independently extracted by two authors. Data analysis was performed using the group subjects were randomised to ('intention to screen'), whether or not they were ever screened. To estimate the effect of Hemoccult screening on colorectal cancer mortality, we calculated relative risks and risk differences for each trial, and then overall, using fixed and random effects models and tested for heterogeneity of effects. We calculated summary measures of effect including all trials and also for just the randomised controlled trials. We also calculated a summary measure of effect, adjusted for attendance at screening in each trial (not shown in Meta-view). MAIN RESULTS: Meta-analysis of mortality results from the randomised controlled trials shows that those allocated to screening had a reduction in colorectal cancer mortality of 16% (RR 0.84, CI: 0.77-0.93). When adjusted for screening attendance in the individual studies, the mortality reduction is 23% (RR 0.77, CI: 0.57-0.89). Overall, if 10 000 people were offered a biennial Hemoccult screening program and two-thirds attended for at least one Hemoccult test, there would be 8.5 deaths (CI: 3.6-13.5) from colorectal cancer prevented over 10 years. However, the screening program would also result in 2 800 participants having at least one colonoscopy, if screening harms from the Minnesota trial are considered, and there would be 3.4 colonoscopy complications (perforation or haemorrhage). If screening harms from the Gothenburg trial are considered, approximately 600 participants would need at least one sigmoidoscopy and double contrast barium enema, resulting in 1.8 perforations or haemorrhages. REVIEWER'S CONCLUSIONS: Screening benefits include reduction in colorectal cancer mortality, possible reduction in cancer incidence through detection and removal of colorectal adenomas and potentially, treatment of early colorectal cancers may involve less invasive surgery. Harmful effects of screening include the physical complications of colonoscopy, disruption to lifestyle, stress and discomfort of testing and investigations, and the anxiety caused by falsely positive screening tests. Although screening benefits are likely to outweigh harms for populations at increased risk of colorectal cancer, we need more information about the harmful effects of screening, the community's responses to screening and screening costs for different health care systems before widespread screening can be recommended. PMID- 10796761 TI - Corticosteroids for the resolution of malignant bowel obstruction in advanced gynaecological and gastrointestinal cancer. AB - BACKGROUND: Gastrointestinal and ovarian cancers are common cancers. The incidence of associated malignant bowel obstruction in patients with advanced cancers of these types is not known, and the best management of these patients is controversial. Inappropriate management may result in uncontrolled (faeculant) vomiting, pain and distress. Management of the symptoms can include palliative surgery, nasogastric tube suction together with intravenous fluids, or pharmacological means, such as corticosteroids. There is uncertainty regarding both the efficacy and possible harmful effects of corticosteroids, and also the most effective type, dose/dosing regime, route and period of administration. OBJECTIVES: To locate, appraise and summarise evidence from scientific studies on intestinal obstruction due to advanced gynaecological and gastrointestinal cancer, in order to assess the efficacy of corticosteroids. SEARCH STRATEGY: A comprehensive list of all studies was provided by an extensive search of the electronic databases, relevant journals, reference lists, the grey literature, contact with investigators and other search strategies outlined in the methods. SELECTION CRITERIA: As the review concentrates on the 'best evidence' available of the role of corticosteroids in malignant bowel obstruction due to advanced gynaecological and gastrointestinal cancer the inclusion criteria were kept fairly broad so as to include all studies relevant to the question DATA COLLECTION AND ANALYSIS: Data extraction forms were used to collect data from the studies included in the review. The data was checked by a secondary searcher to reduce error. A qualitative analysis was performed of the dichotomous data of resolution of obstruction and death at one month, obtained from the randomised controlled trials of corticosteroids versus placebo. Both fixed and random effect models were used. Number needed to treat (NNT) was derived from the odds ratio. Kaplan-Meier survival curves from individual patient data were also analysed. Studies of lower methodological quality were assessed in a qualitative manner. MAIN RESULTS: Three unpublished, randomised, placebo, double blind controlled trials and seven published (prospective and retrospective) trials were considered eligible. Using only the randomised trials, there is a trend, which is not statistically significant, for the resolution of bowel obstruction using corticosteroids. There is no statistically significant difference in mortality at one month, nor in the Kaplan-Meier curves, which describe the survival of patients on corticosteroids or placebo. Number needed to treat is 6 (3, infinity) ie six patients need to be treated with corticosteroids to resolve one episode of bowel obstruction. The results are robust to fixed and random effects models and to 'best' and 'worst case' scenarios on the missing data from patients. The morbidity associated with corticosteroids appears to be very low, though the quality of the data limits this conclusion. No other outcomes were available from the published data or from the authors. REVIEWER'S CONCLUSIONS: There is a trend for evidence that corticosteroids of dose range 6-16 mg dexamethasone given intravenously may bring about the resolution of bowel obstruction. Equally, the incidence of side effects in all the included studies is extremely low. Corticosteroids do not seem to affect the length of survival of these patients. PMID- 10796762 TI - Liquid and fluid agents for preventing adhesions after surgery for subfertility. AB - BACKGROUND: Pelvic surgery is associated with high rates of both de novo adhesion formation and adhesion reformation. Although the role of pelvic and/or tubal surgery in the management of infertility is more limited since the development of in-vitro fertilisation such surgery remains indicated for a number of selected patients. Other forms of pelvic surgery will remain prevalent in women of reproductive age (e.g. endometriosis surgery, ovarian cystectomy, myomectomy). Since subsequent fertility is reduced with increasing severity of periadnexal adhesions, pelvic adhesions will remain a clinical problem in infertility patients. Adjuvant therapy has been promoted for many years to prevent adhesion formation. Numerous substances have been used experimentally in animal models, many have been advocated for use during human surgery, and some are widely used in clinical practice. Steroids and antihistamines are given in the belief that they will promote fibrinolysis during healing without preventing healing. OBJECTIVES: To investigate whether pharmacological and liquid agents used as adjuvants during pelvic surgery in infertility patients lead to a reduction in the incidence or severity of postoperative adhesion (re-)formation, and/or an improvement in subsequent pregnancy rates. SEARCH STRATEGY: The specialised database of the Menstrual Disorders and Subfertility Group. SELECTION CRITERIA: Randomised controlled trials investigating the use of pharmacological and liquid agents to prevent adhesion formation after pelvic surgery for infertility. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the first 2 authors. Differences of opinion were registered and resolved by consensus with the senior author (RJL). Two by two tables were generated for each trial for the dichotomous outcome of pregnancy and the effects on pregnancy rate of each study is expressed as an odds ratio with 95% confidence intervals. MAIN RESULTS: None of the pharmacological or liquid agents investigated in a randomised controlled fashion was shown to improve postoperative pregnancy rates. There was some evidence that steroids reduced the incidence and severity of postoperative adhesion formation. Dextran appeared to neither reduce the incidence or severity of adhesions. REVIEWER'S CONCLUSIONS: The routine use of pharmacological agents to prevent post-operative adhesions after infertility surgery cannot be recommended on the basis of the available evidence derived from RCTs. In connection with adhesion prevention, the evidence with regard to steroids is far from perfect but tentatively suggests that they may be beneficial. Further randomised studies should be conducted to investigate this further. PMID- 10796763 TI - Gonadotropin releasing hormone agonist protocols for pituitary desensitization in in vitro fertilization and gamete intrafallopian transfer cycles. AB - BACKGROUND: Gonadotropin releasing hormone agonists (GnRHa) are used in assisted reproduction cycles to reversibly block pituitary function and prevent a luteinizing hormone surge. In the short and ultrashort protocols of GnRHa administration, injection of gonadotropins is commenced a few days after the start of GnRHa. In the long protocols (with GnRHa started either in the midluteal phase or in the early follicular phase) gonadotropin administration is delayed until pituitary desensitization has been achieved, usually 2-3 weeks. OBJECTIVES: To conduct a systematic overview of available data comparing short or ultrashort and long GnRHa protocols for pituitary desensitization in in vitro fertilization (IVF) and gamete intra-fallopian transfer (GIFT) treatment cycles. SEARCH STRATEGY: Search strategies included on-line searching of the MEDLINE and EMBASE data bases and the Cochrane Menstrual Disorders and Subfertility Group's Specialised Register from 1982 to 1998, and hand searching of bibliographies of relevant publications and reviews, and abstracts of scientific meetings. SELECTION CRITERIA: Randomized trials of short or ultrashort versus long (follicular or luteal phase start) GnRHa protocols in IVF or GIFT treatment cycles. DATA COLLECTION AND ANALYSIS: Data were extracted into 2 x 2 tables. For the primary outcome, clinical pregnancy per cycle started, the overall common odds ratio (OR) and the risk difference with 95% confidence interval (CI) were calculated after verifying the presence of homogeneity of treatment effect across all trials. The following subgroup comparisons were performed: ultrashort versus long protocols, short versus long protocols and, within each of these comparisons, subgroups of studies which used the long protocol with follicular phase start or the long protocol with luteal phase start. Secondary outcomes considered were clinical pregnancy per oocyte retrieval and per embryo transfer, spontaneous abortion, ongoing/delivered pregnancy per cycle started, number of ampoules of gonadotropin used, number of oocytes retrieved, and fertilization rate. MAIN RESULTS: Twenty-six trials met the inclusion criteria. The common OR for clinical pregnancy per cycle started was 1.32 (95% CI, 1.10 - 1.57) in favour of the long GnRHa protocol. The studies were subgrouped, depending on whether, in the long protocol, the GnRHa was commenced in the follicular phase (8 trials) or luteal phase (16 trials). The respective ORs were 1.54 (95% CI, 1.11 - 2. 13) and 1.21 (95% CI, 0.98 - 1.51). After excluding the four trials using the ultrashort protocol, the OR for long versus short protocols (22 trials) was 1.27 (95% CI, 1.04 - 1.56). A comparison of long versus ultrashort protocols (4 trials) produced an OR of 1. 47 (95% CI, 1.02 - 2.12). REVIEWER'S CONCLUSIONS: On the basis of clinical pregnancy rate per cycle started, this meta-analysis demonstrates the superiority of the long protocol over the short and ultrashort protocols for GnRHa use in IVF and GIFT cycles. PMID- 10796764 TI - Intra-cytoplasmic sperm injection versus partial zona dissection, subzonal insemination and conventional techniques for oocyte insemination during in vitro fertilisation. AB - BACKGROUND: In vitro fertilisation (IVF) and embryo transfer as treatment for male factor infertility is associated with lower fertilisation and pregnancy rates than for other indications. Since the late 1980s several assisted fertilisation techniques have emerged and have been rapidly developed to try to enhance results for couples with male factor infertility, or to help couples with severe male factor for whom conventional IVF was not possible. The technique of partial zona dissection (PZD) was developed to increase the probability that a sperm capable of fertilisation comes in contact with the oocyte. Although this method improved conventional IVF results, the improvement was only marginal and relatively large numbers of sperm are still required. This drawback applied less to the subsequent technique of subzonal microinjection of spermatozoa into the perivitelline space (SUZI). However, for all of these techniques fertilisation rates remained low, rates of polyspermic fertilisation were increased, and cases with a very limited number of spermatozoa in the ejaculate could still not be treated. The advent of intra-cytoplasmatic sperm injection (ICSI) of a single sperm (or sperm head or nucleus) into the oocyte appears to be an important breakthrough. OBJECTIVES: To investigate whether ICSI improves fertilisation and/or pregnancy rates in comparison to other fertilisation techniques. SEARCH STRATEGY: The Menstrual Disorders and Subfertility Group search strategy (see Review Group details) was used to identify trials that had compared ICSI with other infertility techniques, such as PZD, SUZI, conventional IVF and additional IVF. SELECTION CRITERIA: Trials were included if they compared the effects of these techniques on fertilisation and pregnancy outcomes. Only randomised studies were included in this review. DATA COLLECTION AND ANALYSIS: Ten studies met the inclusion criteria for this review. Eight studies compared ICSI with conventional IVF. One study compared ICSI with SUZI and one study compared ICSI with additional IVF. Data was extracted independently by two reviewers. Where relevant data was missing or unclear, the authors had been consulted. Male participants were classified according to their semen parameters, i.e. normal semen (concentration >20 million per ml, motility >50%, morphology >14%), borderline semen (concentration 10-20 million per ml, motility 30-50%, morphology 4-14% normal forms) and very poor semen (concentration <10 million per ml, motility <30%, morphology <4% normal forms). MAIN RESULTS: For couples with normal semen there is no evidence of a difference in fertilisation rates per retrieved oocyte or pregnancy rates between ICSI and conventional IVF. On the other hand, for fertilisation rate per inseminated oocyte, ICSI appears to result in better outcomes than IVF for normal semen. For couples with borderline semen ICSI results in higher fertilisation rates (all) than IVF. Couples with very poor semen will have better fertilisation outcomes with ICSI than with SUZI or additional IVF. REVIEWER'S CONCLUSIONS: There is evidence from this systematic review that fertilisation rates are significantly better with ICSI than IVF in couples with borderline semen. When the semen parameters are normal there is insufficient evidence of a difference in effectiveness between ICSI and IVF when retrieved oocytes were the unit of randomisation. However, there was a small but statistically significant increase in fertilisation rate when inseminated oocytes were the unit of randomisation. Total fertilisation rates were significantly reduced in ICSI cycles than IVF but there were no damaged oocytes in IVF cycles regardless of the semen parameters. PMID- 10796765 TI - Intra-venous albumin for preventing severe ovarian hyperstimulation syndrome. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic condition that occurs after the administration of human menopausal gonadotrophin (hMG) with or without gonadotrophin releasing hormone (GnRH) agonists. OHSS is a threat to every woman undergoing ovulation induction and is potentially lethal in its severest form. Severe OHSS is characterised by growth of multiple large follicles with massive extravascular protein rich fluid shift. This may lead to hypovolaemia, haemoconcentration, oliguria, and electrolyte disturbance. Human albumin solutions are now used in the management of shock and other conditions in which restoration of blood volume is urgent, the acute management of burns, and clinical situations associated with hypoproteinaemia. Recently, a number of clinical trials with conflicting results have been reported in which albumin has been tested as a possible way for preventing the severe form of OHSS. OBJECTIVES: To review the effectiveness of human albumin administration in prevention of severe ovarian hyperstimulation syndrome. SEARCH STRATEGY: The Menstrual Disorders and Subfertility Group literature search strategy was used to identify randomised trials that had compared the use of human albumin with placebo or no treatment in the prevention of severe ovarian hyperstimulation syndrome. A diverse search strategy was employed, including handsearching of core journals from 1966 to the present, searching bibliographies of relevant trials, MEDLINE, EMBASE, PsychLIT and CINAHL databases, the MDSG specialised register, abstracts from North American and European meetings and contact with authors of relevant papers. SELECTION CRITERIA: Trials were included if they compared the effect of human albumin with placebo or no treatment on relevant outcomes. Only randomised controlled studies were included in this review. DATA COLLECTION AND ANALYSIS: Trials under consideration were evaluated for methodological quality and appropriateness for inclusion without consideration of their results. Relevant data were extracted independently by two reviewers using the standardized data extraction sheet. Validity was assessed in terms of method of randomization, completeness of follow-up, presence or absence of crossover and co-intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were calculated using the Peto modified Mantel-Haenszel technique. MAIN RESULTS: Meta-analysis of the three included trials demonstrated significant reduction in severe ovarian hyperstimulation syndrome on administration of human albumin (common odds ratio 0.1, 95% confidence interval 0.03 to 0.39). There was no evidence of an increase in the pregnancy rate (common odds ratio 1.69, 95% confidence interval 0.7 to 4.07). REVIEWER'S CONCLUSIONS: This review shows a clear benefit from administration of intra-venous albumin at the time of oocyte retrieval in prevention of severe OHSS in high-risk cases. However, the results of this review can not be regarded as conclusive as they are based on only three small trials. Further trials are urgently needed. PMID- 10796766 TI - Dietary interventions for phenylketonuria. AB - BACKGROUND: Phenylketonuria is an inherited disease for which the main treatment is the dietary restriction of the amino acid phenylalanine. The diet has to be initiated in the neonatal period to prevent or reduce mental handicap however the diet is very restrictive and unpalatable and can be difficult to follow. Whether the diet can be relaxed or discontinued during adolescence or should be continued for life remains a contraversial issue which we aim to address in this review. OBJECTIVES: To assess the effects of a phenylalanine restricted diet commenced early in life for patients with phenylketonuria. To assess the possible adverse effects of relaxation or termination of the diet on intelligence, neuropsychological outcomes and mortality, and to assess the effect on growth, nutritional status and eating behaviour and quality of life. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Trials Register which is a specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Additional studies were identified from handsearching the Journal of Inherited Metabolic Disease (from inception, 1978 to 1998). The manufacturers of dietary products for phenylketonuria were also contacted. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised controlled trials comparing a phenylalanine restricted diet to either relaxation or termination of dietary restrictions in patients with phenylketonuria. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the trial eligibility, methodological quality and extracted the data. MAIN RESULTS: Four studies were included in this review including a total of 251 patients. Few statistically significant differences were found between treatment and comparison groups for any of the outcomes apart from for blood phenylalanine level and intelligence quotient. Blood phenylalanine levels were significantly lower in those subjects with phenylketonuria following a phenylalanine restricted diet compared to those on a less restricted or relaxed diet (weighted mean difference at three months -672.203, 95% Confidence interval (CI) -813.799 to - 530.608). Intelligence quotient was significantly higher in subjects who continued on the phenylalanine restricted diet compared to those who terminated the diet (weighted mean difference after 12 months -5.00, 95% CI -9.595 to 0.405). However this is based on the results of only one study. REVIEWER'S CONCLUSIONS: The results of non-randomised studies have concluded that a phenylalanine restricted diet is effective in reducing blood phenylalanine levels and improving intelligence quotient and neuropsychological outcome. No randomised controlled trials have assessed the effect of a phenylalanine restricted diet versus no dietary restrictions from diagnosis. In view of evidence from non randomised studies, such a trial would now be unethical and it is recommended that phenylalanine restricted diet should be commenced at the time of diagnosis. There is uncertainty about the precise level of phenylalanine restriction and when, if ever, the diet should be relaxed. These questions should be addressed by randomised controlled trials with careful consideration given to which patients to include. PMID- 10796767 TI - Antibiotics for treating leptospirosis. AB - BACKGROUND: Leptospirosis is a parasitic disease transmitted by animals. Severe leptospirosis may result in hospitalisation and about five per cent of the patients die. In clinical practice, penicillin is widely used for treating leptospirosis. OBJECTIVES: To evaluate the effectiveness and safety of antibiotics versus placebo or other antibiotic regimens in treating leptospirosis. We addressed the following clinical questions: a) Are treatment regimens with antibiotics more efficient than placebo for leptospirosis? b) Are treatment regimens with antibiotics safe when compared to placebo for leptospirosis? c) Which antibiotic regimen is the most efficient and safest in treating leptospirosis? SEARCH STRATEGY: Electronic searches and searches of the identified articles were combined. SELECTION CRITERIA: STUDIES: Randomised clinical trials in which antibiotics were used as treatment for leptospirosis. Language, date, or other restrictions were not applied. PARTICIPANTS: Patients with clinical manifestations of leptospirosis. INTERVENTIONS: Any antibiotic regimen compared with a control group (placebo or another antibiotic regimen). DATA COLLECTION AND ANALYSIS: Data and methodological quality of each trial were independently extracted and assessed by two reviewers. The random effects model was used irrespective of significant statistical heterogeneity. MAIN RESULTS: Three trials met inclusion criteria. Allocation concealment and double blind methods were not clearly described in two. Of the patients enrolled, 75 were treated with placebo and 75 with antibiotics: 61 (81.3%) penicillin and 14 (18.6%) doxycycline. The patients assigned to antibiotics compared to placebo showed: a) Mortality: 1% (1/75) versus 4% (3/75); risk difference -2%, 95% confidence interval -8% to 4%. b) Duration of hospital stay (days): weighted mean difference 0.30, 95% confidence interval -1.26 to 1.86. c) Prolonged hospital stay (> seven days): 30% (7/23) versus 74% (14/19); risk difference -43%, 95% confidence interval -70% to -16%. Number needed-to-treat 3, 95% confidence interval 2 to 7. d) Period of disappearance of fever (days): weighted mean difference -4.04, 95% confidence interval -8.65 to 0.58. e) Leptospiruria: 5% (4/75) versus 40% (30/75); risk difference -46%, 95% confidence interval -88% to 3%. Number needed-to-treat 2, 95% confidence interval 1 to 33. REVIEWER'S CONCLUSIONS: Antibiotic regimens for treatment of leptospirosis is a form of care for which the evidence is insufficient to provide clear guidelines for practice. The randomised trials suggest that antibiotics could be a useful treatment for leptospirosis. Because of the questionable quality of two of the three trials, the indication for general use of antibiotics is uncertain. However, the evidence suggest that penicillin may cause more good than harm. PMID- 10796768 TI - Bladder training for urinary incontinence in adults. AB - BACKGROUND: Bladder training is widely used for the treatment of urinary incontinence. It is generally used for the treatment of people with urge incontinence or detrusor instability, although it is also thought that it might be of use for people with mixed incontinence or stress incontinence. OBJECTIVES: To assess the effects of bladder training for the treatment of urinary incontinence. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register up to July 1999. Date of the most recent search: July 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials of bladder training for the treatment of incontinence. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data that were then cross-checked by the third reviewer. All three reviewers assessed trial quality. MAIN RESULTS: We found seven eligible trials with a total of 259 predominantly female patients with urinary urge incontinence. The quality of trials was variable. Three trials involving 92 women compared bladder training with no bladder training. These tended to favour bladder training but data were available for only a limited number of pre specified outcomes that varied across the three trials. No data describing long term follow up are available. One trial compared bladder training with drug therapy, but was inconclusive. Another trial compared bladder training with an electronic prompt device. The trial was small and included only 20 women. Data were not presented in a form suitable for quantitative analysis. Two further trials compared bladder training supplemented by drug therapy with bladder training alone. Again, the limited data provided insufficient evidence on which to draw conclusions. REVIEWER'S CONCLUSIONS: Bladder training may be helpful for the treatment of urinary urge incontinence, but this conclusion can only be tentative, based on the evidence available. There was not enough evidence to show whether drug therapy was better than bladder training or useful as a supplement to it. PMID- 10796769 TI - Electrical stimulation for faecal incontinence in adults. AB - BACKGROUND: Faecal incontinence is a particularly embarrassing and distressing condition with significant medical, social and economic implications. Electrical stimulation has been used with apparent success in the treatment of faecal incontinence. However, standards of treatment are still lacking and the magnitude of alleged benefits has yet to be established. OBJECTIVES: To determine the effects of electrical stimulation for the treatment of faecal incontinence in adults. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of potentially eligible articles up to November 1999. Date of the most recent searches: November 1999. SELECTION CRITERIA: All randomised or quasi randomised trials evaluating electrical stimulation in adults with faecal incontinence. DATA COLLECTION AND ANALYSIS: Three reviewers assessed the methodological quality of potentially eligible trials and two reviewers independently extracted data from the included trial. A wide range of outcome measures were considered. MAIN RESULTS: Only one eligible trial with 40 participants was identified. It was a randomised trial, but it suffered from methodological drawbacks and did not follow up patients beyond the end of the trial period. Findings from this trial suggest that electrical stimulation with anal biofeedback and exercises provides more short-term benefits than vaginal biofeedback and exercises for women with obstetric-related faecal incontinence. No further conclusions could be drawn from the data available. REVIEWER'S CONCLUSIONS: At present, there are insufficient data to allow reliable conclusions to be drawn on the effects of electrical stimulation in the management of faecal incontinence. There is a suggestion that electrical stimulation may have a therapeutic effect, but this is not certain. Larger, more generalisable trials are needed. PMID- 10796770 TI - Antidepressants for depression in people with physical illness. AB - OBJECTIVES: To determine whether antidepressants are clinically effective and acceptable for the treatment of depression in people who also have a physical illness. SEARCH STRATEGY: Medline, Cochrane Library Trials Register and Cochrane Depression and Neurosis Group Trials Register were all systematically searched, supplemented by hand searches of two journals and reference searching. SELECTION CRITERIA: All relevant randomised trials comparing any antidepressant drug (as defined in the British National Formulary) with placebo or no treatment, in patients of either sex over 16, who have been diagnosed as depressed by any criterion, and have a specified physical disorder (for example cancer, myocardial infarction). "Functional" disorders where there is no generally agreed physical pathology (e.g. irritable bowel syndrome) were excluded. The main outcome measures are numbers of individuals who recover/improve at the end of the trial and, as a proxy for treatment acceptability, numbers who complete treatment. DATA COLLECTION AND ANALYSIS: Data was extracted independently by the reviewers onto data collection forms and differences settled by discussion. MAIN RESULTS: 18 studies were included, covering 838 patients with a range of physical diseases (cancer 2, diabetes 1, head injury 1, heart 1, HIV 5, lung 1, multiple sclerosis 1, renal 1, stroke 3, mixed 2). Depression was diagnosed clinically in 3 studies, otherwise by structured interview or checklist. Only 5 studies described how they performed randomisation. 1 study compared drug with no treatment, and the rest with placebo: all of the latter said they were double blind. 6 studies used SSRIs, 3 atypical antidepressants, and the remainder tricyclics. Patients treated with antidepressants were significantly more likely to improve than those given placebo (13 studies, OR 0.37, 95% CI 0.27-0.51) or no treatment (1 study, OR 3.45, 95% CI 11.1-1.10). About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Most antidepressants (tricyclics and SSRIs together, 15 trials ) produced a small but significant increase in dropout (OR 1.66, 95% CI 1.14-2.40. NNH 9.8, 95% CI 5.4 42.9). The "atypical" antidepressant mianserinproduced significantly less dropout than placebo. Only 2 studies used numerical scales designed to measure effects on function and quality of life; in HIV (Karnofsky scale), drug was better than no treatment; in lung disease (Sickness Impact Profile), drug was not significantly different from placebo. Only 7 studies reported looking for changes in the physical disease. Antidepressants produced no change in immune function in HIV relative to placebo (2 studies) or no treatment (1 study). Relative to placebo, antidepressants produced no change in cardiovascular function in heart disease, in respiratory function in lung disease, or in vital signs or laboratory tests in cancer (1 study each). Nortriptyline produced worse control in diabetes. Trends towards tricyclics being more effective than SSRIs, but also more likely to produce dropout were noted, but these are based on non-randomised comparisons between trials. REVIEWER'S CONCLUSIONS: The review provides evidence that antidepressants, significantly more frequently than either placebo or no treatment, cause improvement in depression in patients with a wide range of physical diseases. About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Antidepressants seem reasonably acceptable to patients, in that about 10 patients would need to be treated with antidepressants to produce one dropout from treatment which would not have occurred had they been given placebo (NNH 9.8, 95% CI 5.4-42.9). (ABSTRACT TRUNCATED) PMID- 10796771 TI - Surgery for cervical intraepithelial neoplasia. AB - OBJECTIVES: Cervical intra-epithelial neoplasia is treated by local ablation or lower morbidity excision techniques. Choice of treatment depends on the severity of the disease. The objective of this review was to assess the effects of alternative surgical treatments for cervical intra-epithelial neoplasia. SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Group trials register and MEDLINE up to July 1997. SELECTION CRITERIA: Randomised and quasi-randomised trials of alternative surgical treatments in women with cervical intra-epithelial neoplasia. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and two reviewers abstracted data independently. MAIN RESULTS: Twenty-three trials were included. Seven surgical techniques were tested in various comparisons. No significant difference in eradication of disease was shown, other than between laser ablation and loop excision. This was based on one trial where the quality of randomisation was doubtful. Large loop excision of the transformation zone appeared to provide the most reliable specimens for histology. Morbidity was lower than with laser conisation, although all five trials did not provide data for every outcome. There were not enough data to assess the effect on morbidity compared with laser ablation. REVIEWER'S CONCLUSIONS: The evidence suggests that there is no obviously superior surgical technique for treating cervical intra epithelial neoplasia. PMID- 10796772 TI - Colloid solutions for fluid resuscitation. AB - BACKGROUND: Colloids are widely used in the replacement of fluid volume, however doubts remain as to their benefits. Different colloids vary in their molecular weight and therefore in the length of time they remain in the circulatory system. Because of this and their other characteristics, they may differ in their safety and efficacy. OBJECTIVES: To compare the effects of different colloid solutions in patients thought to need volume replacement. SEARCH STRATEGY: The Cochrane Injuries Group specialised register, The Cochrane Controlled trials register (all years), MEDLINE (1994-98), EMBASE (1974-98) were searched. Bibliographies of trials retrieved were searched and drug companies manufacturing colloids were contacted for information. SELECTION CRITERIA: Randomised and quasi randomised trials comparing colloid solutions in critically ill and surgical patients thought to need volume replacement. The main outcomes measured were death, amount of whole blood transfused and incidence of adverse reactions. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data and assessed the quality of the trials. MAIN RESULTS: 46 trials met the inclusion criteria, with a total of 2884 participants. Many of the trials were small. In the majority of trials quality was poor or uncertain. Deaths were obtained from 27 trials. Twenty three trials recorded the amount of blood transfused, however quantitative analysis was not possible due to skewness and variable reporting. Thirteen trials recorded adverse reactions, but none occurred. For albumin or PPF versus hydroxyethyl starch (HES) 20 trials reported mortality. The pooled relative risk (RR) was 1.17 (95% CI 0.91, 1.50). For albumin or PPF versus gelatin 3 trials reported mortality. The RR was 0.99 (0.69, 1.42). For gelatin vs HES 3 trials reported mortality, RR was 0.97 (0.65, 1.44). RR was not estimable in the albumin vs dextran, gelatin vs dextran, and HES vs dextran groups. REVIEWER'S CONCLUSIONS: From this review, there is no evidence that one colloid solution is more effective or safe than any other, although the confidence intervals are wide and do not exclude clinically significant differences between colloids. Larger trials of fluid therapy are needed if clinically significant differences in mortality are to be detected or excluded. PMID- 10796773 TI - Cytomegalovirus prophylaxis with antiviral agents for solid organ transplantation. AB - OBJECTIVES: To assess the efficacy of antiviral agents in solid organ transplant recipients in the prevention of cytomegalovirus infection and symptomatic disease and in the reduction of the incidence of acute rejection, graft loss and death. SEARCH STRATEGY: A computerised search was conducted on Medline, Embase and Pascal. The reference lists of the current review articles and some congress proceedings were searched manually (Transplantation Proceedings, American Thoracic Society, European Society of Organ Transplantation). SELECTION CRITERIA: Prospective, randomised studies in adults or paediatric recipients of a solid organ transplant, in which one arm received a prophylactic treatment with acyclovir and/or ganciclovir, started before cytomegalovirus infection, and the control arm received placebo or no treatment. DATA COLLECTION AND ANALYSIS: Data were extracted from each trial and a letter sent to the authors to ask them to verify the data extracted, and to provide any data that was missing. For each outcome, several methods were used to calculate the chi-square for association and the estimate for the treatment effect with its 95% confidence interval, with an additive model (rate difference), or a multiplicative model (odds ratio, relative risk). We considered the test of association to be significant when the p value was less than 0.01 and the homogeneity test to be significant when the p value was less than 0.1. MAIN RESULTS: Prophylactic treatment was found to be associated with a significant decrease in cytomegalovirus disease compared with placebo or no treatment, using the logarithm of relative risk method (relative risk 0.51, 95% confidence interval 0. 41-0.64, p value for X(2) association < 0.001). Prophylactic treatment also decreased the rate of cytomegalovirus infection (RR 0. 62, 95%CI 0.53-0.73, p < 0.001). Our analysis failed to show a significant decrease in graft loss, acute rejection or death in the prophylactic treatment group. Sub-group analysis based on the type of antiviral agent (acyclovir or ganciclovir) and on the type of organ (kidney or liver) gave comparable results. REVIEWER'S CONCLUSIONS: The use of antiviral agents for the prevention of cytomegalovirus disease and cytomegalovirus infection in solid organ transplantation is supported by this meta-analysis. PMID- 10796774 TI - Cranberries for preventing urinary tract infections. AB - BACKGROUND: Cranberries (particularly in the form of cranberry juice) have been used widely for several decades for the prevention and treatment of urinary tract infections. The aim of this review is to assess the effectiveness of cranberries in preventing such infections. OBJECTIVES: To assess the effectiveness of cranberry juice and other cranberry products in preventing urinary tract infections in susceptible populations. SEARCH STRATEGY: Electronic databases and the Internet were searched using English and non English language terms; companies involved with the promotion and distribution of cranberry preparations were contacted; reference lists of review articles and relevant trials were searched. SELECTION CRITERIA: All randomised or quasi randomised controlled trials of cranberry juice/products for the prevention of urinary tract infections in susceptible populations. Trials of men, women or children were included. DATA COLLECTION AND ANALYSIS: Reviewers RJ and LM independently assessed and extracted information using specially designed data extraction forms. For each included trial, information was collected on methods of the trial, participants, interventions and outcomes. We were unable to perform statistical analysis due to the nature of the data available for review. MAIN RESULTS: Four trials met the inclusion criteria (three cross-over, one parallel group). Three compared the effectiveness of cranberry juice versus placebo juice or water and one compared the effectiveness of cranberry capsules versus placebo. Two further trials were excluded. The outcomes of interest were number of urinary tract infections in each group (symptomatic and asymptomatic), side effects and adherence to therapy. Data from three out of the four trials indicated that cranberries were effective for at least one of the outcomes of interest. The quality of the four included trials was poor, however, and thus the reliability of the results must be questionable. REVIEWER'S CONCLUSIONS: The small number of poor quality trials gives no reliable evidence of the effectiveness of cranberry juice and other cranberry products. The large number of dropouts/withdrawals from the trials indicates that cranberry juice may not be acceptable over long periods of time. Other cranberry products such as cranberry capsules may be more acceptable. On the basis of the available evidence, cranberry juice cannot be recommended for the prevention of urinary tract infections in susceptible populations. Further properly designed trials with relevant outcomes are needed. PMID- 10796775 TI - Cranberries for treating urinary tract infections. AB - BACKGROUND: Cranberries (particularly in the form of cranberry juice) have been used widely for several decades for the prevention and treatment of urinary tract infections. The aim of this review is to assess the effectiveness of cranberries in treating such infections. OBJECTIVES: To assess the effectiveness of cranberries for the treatment of urinary tract infections. SEARCH STRATEGY: The search strategy developed by the Cochrane Renal Group was used. Also, companies involved with the promotion and distribution of cranberry preparations were contacted; electronic databases and the Internet were searched using English and non English language terms; reference lists of review articles and relevant trials were also searched. SELECTION CRITERIA: All randomised or quasi randomised controlled trials of cranberry juice or cranberry products for the treatment of urinary tract infections. Trials of men, women or children were included. DATA COLLECTION AND ANALYSIS: Titles and abstracts of studies that were potentially relevant to the review were screened by one reviewer, RJ, who discarded studies that were clearly ineligible but aimed to be overly inclusive rather than risk losing relevant studies. Reviewers RJ and LM independently assessed whether the studies met the inclusion criteria. Further information was sought from the authors where papers contained insufficient information to make a decision about eligibility. MAIN RESULTS: No trials were found which fulfilled all of the inclusion criteria. Two trials were excluded because they did not have any relevant outcomes. REVIEWER'S CONCLUSIONS: After a thorough search, no randomised trials which assessed the effectiveness of cranberry juice for the treatment of urinary tract infections were found. Therefore, at the present time, there is no good quality evidence to suggest that it is effective for the treatment of urinary tract infections. Well-designed parallel group, double blind trials comparing cranberry juice and other cranberry products versus placebo to assess the effectiveness of cranberry juice in treating urinary tract infections are needed. Outcomes should include reduction in symptoms, sterilisation of the urine, side effects and adherence to therapy. Dosage (amount and concentration) and duration of therapy should also be assessed. Consumers and clinicians will welcome the evidence from these trials. PMID- 10796776 TI - Interventions for emergency contraception. AB - OBJECTIVES: To determine which emergency contraceptive method following unprotected intercourse is the most effective, safe and convenient for use in preventing pregnancy. SEARCH STRATEGY: The search strategy included electronic searches of the Cochrane Controlled Trials Register, Popline, Chinese biomedical databases and HRP emergency contraception database. In addition, references of retrieved papers were searched and researchers in the field and two pharmaceutical companies were contacted. SELECTION CRITERIA: Randomized or quasi randomized studies including women attending services for emergency contraception following a single act of unprotected intercourse were eligible. DATA COLLECTION AND ANALYSIS: Data on outcomes and trial characteristics were extracted in duplicate by two reviewers. Results were expressed as relative risk using a fixed effects model with 95 % confidence interval. MAIN RESULTS: Fifteen trials were included in the review. The majority (8/15) of the trials were conducted in China. Most comparisons between different interventions included one or two trials although some trials were appropriately sized with power calculations. Levonorgestrel appears to be more effective than Yuzpe regimen (2 trials, RR: 0.51, 95 % CI: 0.31-0.84) and causes less side-effects (RR: 0.80, 95 % CI: 0.76 to 0.84). Levonorgestrel was less effective than locally manufactured mifepristone in a single, large Chinese study (RR: 2.17, 95 % CI: 1.00 to 4.77). Effectiveness of different doses of mifepristone seem to be similar but the frequency of delay in onset of the subsequent menstrual period increases with increased dose. REVIEWER'S CONCLUSIONS: Levonorgestrel and mifepristone seem to offer the highest efficacy with an acceptable side-effect profile. One disadvantage of mifepristone is that it causes delays in onset of subsequent menses which may induce anxiety. However, this seems to be dose-related and low doses of mifepristone minimise this side-effect without compromising effectiveness. Future studies should compare the effectiveness of mifepristone with levonorgestrel. PMID- 10796777 TI - Antibiotic prophylaxis for intrauterine contraceptive device insertion. AB - BACKGROUND: Concern about the risk of upper genital tract infection (pelvic inflammatory disease) often limits use of the IUD, a highly effective contraceptive. Prophylactic antibiotic administration around the time of induced abortion significantly reduces the risk of postoperative endometritis.(Sawaya, 1996) Since the risk of IUD-related infection is limited to the first few weeks to months after insertion,(Lee, 1983; Farley, 1992) contamination of the endometrial cavity at the time of insertion(Mishell, 1966) appears to be the mechanism, rather than the IUD or string itself. Thus, antibiotic administration before IUD insertion might reduce the risk of upper genital tract infection from passive introduction of bacteria at insertion. OBJECTIVES: To assess the effectiveness of prophylactic antibiotic administration before IUD insertion in reducing IUD-related complications and discontinuations within three months of insertion. The primary outcome was pelvic inflammatory disease (four reports) or early removals of the device (two reports). SEARCH STRATEGY: We searched both MEDLINE and EMBASE, handsearches of journals through CENTRAL, and lists of references. We also wrote to international experts in the field to identify unpublished studies. SELECTION CRITERIA: We included randomized controlled trials using any antibiotic compared with a placebo. We found four such trials; two had pilot study data available. DATA COLLECTION AND ANALYSIS: DATA EXTRACTION: We used searches of MEDLINE, EMBASE, and handsearches of journals available through CENTRAL. We also reviewed lists of references in original research and in review articles. We wrote to experts to identify unpublished trials and made telephone calls to authors to supply missing information. Two independent reviewers abstracted data. We assessed the validity of each study using methods suggested in the Cochrane Handbook. DATA SYNTHESIS: We generated 2x2 tables for the principal outcome measures. We used the Peto modified Mantel-Haenszel technique to calculate odds ratios and assessed statistical heterogeneity between studies. MAIN RESULTS: The odds ratios for pelvic inflammatory disease associated with use of prophylactic doxycycline or azithromycin compared with placebo or no treatment was 0.89 (95%CI 0.53-1.51). Use of prophylaxis was associated with a small reduction in unscheduled vists to the provider (OR 0.82; 95% CI 0.70-0.98). Use of doxycycline or azithromycin had little effect on the likelihood of removal of the IUD within 90 days of insertion (OR 1.05; 95% CI 0.68-1.63). Significant heterogeneity did not exist between studies. REVIEWER'S CONCLUSIONS: Use of either doxycycline 200 mg or azithromycin 500 mg by mouth before IUD insertion confers little benefit. While the reduction in unscheduled visits to the provider was marginally significant, the cost-effectiveness of routine prophylaxis remains questionable. A uniform finding in these trials was the low risk of IUD associated infection, with or without use of antibiotic prophylaxis. PMID- 10796778 TI - Minilaparotomy and endoscopic techniques for tubal sterilisation. AB - OBJECTIVES: To evaluate laparoscopic tubal sterilisation, as compared to minilaparotomy in terms of operative morbidity and mortality. Trials comparing laparoscopy or minilaparotomy with culdoscopy were also included in the review. Different methods used to interrupt tubal patency (excision, occlusion and coagulation) and comparison of different forms of anaesthesia will be considered in different reviews. SEARCH STRATEGY: Randomised controlled trials (RCTs) have been identified by using the search strategy of the Cochrane Collaboration. The Cochrane Controlled Trials Register has been searched. Reference lists of identified trials have been searched. SELECTION CRITERIA: All randomised controlled trials comparing laparoscopy, minilaparotomy and/or culdoscopy for tubal sterilisation. Except in one trial [Taner 1994] where 4 women underwent curettage at the same time, all women requested tubal sterilisation as an interval procedure. DATA COLLECTION AND ANALYSIS: Trials under consideration were evaluated for methodological quality and appropriateness for inclusion. Data were extracted independently by the reviewers. Results are reported as odds ratio for dichotomous outcomes and weighted mean differences for continuous outcomes. MAIN RESULTS: Minilaparotomy vs laparoscopy: There was no difference in major morbidity between the 2 groups. Minor morbidity was significantly less in the laparoscopy group (Peto OR 1.89; 95% CI 1.38, 2.59). Duration of operation was about 5 minutes shorter in the laparoscopy group (WMD 5.34; 95% CI 4. 52, 6.16). Minilaparotomy vs culdoscopy: Women undergoing culdoscopy had more major morbidity than women for whom minilaparotomy was performed (Peto OR 0.14; 95% CI 0.02, 0.98). Duration of operation was about 5 minutes shorter in women undergoing culdoscopy (WMD 4. 91; 95% CI 3.82, 6.01). Laparoscopy vs culdoscopy: In the one trial comparing the two interventions there were no significant differences between the groups with regard to major morbidity. Significantly more women suffered from minor morbidities in the culdoscopy group compared to the laparoscopy group (Peto OR 0.20; 95% CI 0.05, 0.77). REVIEWER'S CONCLUSIONS: Major morbidity seems to be a rare outcome for both, laparoscopy and minilaparotomy. The included studies had limited power to demonstrate significant differences especially for the relatively rare but potentially serious outcomes. Personal preference of the woman and/or of the surgeon can guide the choice of technique. Practical aspects (e.g. cost, maintenance, and sterilisation of the instruments) must be taken into account before implementing the more sophisticated endoscopic techniques in settings with limited resources. Culdoscopy is not recommended as it carries a higher complication rate. PMID- 10796779 TI - Naltrexone maintenance treatment for opioid dependence. AB - BACKGROUND: Despite widespread use of naltrexone maintenance in many countries for more than ten years now (e.g., USA since 1984, UK since 1988) a sound documentation of the research on this drug is still missing. OBJECTIVES: To evaluate the effects of naltrexone maintenance treatment in preventing relapse in opioid addicts after detoxification. SEARCH STRATEGY: We searched MEDLINE, EMBASE, CCTR and handsearched the "Bolletino per le Farmacodipendenze e l'Alcolismo"; contact was sought with pharmaceutical producers of naltrexone, with authors and other CRGs; references of obtained studies. Trials were reliably identified and data extracted. Date of most recent searches: June 1998. SELECTION CRITERIA: All studies controlled for naltrexone; treatment of heroin addicts after detoxification with naltrexone. Studies were classified into three categories (high, moderate or low risk of bias) according to their methodological quality. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed outcome measures taking into consideration adherence to and success of the study intervention. Data was extracted and analysed stratifying for the three categories of study quality. Where possible, meta-analysis was performed. MAIN RESULTS: Eleven studies were included in this review, and not all of them were randomised. Meta-analysis could be done to a low degree only, because the studies and their outcomes were very heterogeneous. The result of this quantitative analysis was statistically poor, and so was the methodological quality of the included studies. REVIEWER'S CONCLUSIONS: The available trials do not allow a final evaluation of naltrexone maintenance treatment yet. A trend in favour of treatment with naltrexone was observed for certain target groups, as described in the literature before. A well-done clinical trial is needed in order to get better evidence as soon as possible. PMID- 10796780 TI - Vaccines for preventing infection with Pseudomonas aeruginosa in people with cystic fibrosis. AB - OBJECTIVES: To assess the effectiveness of vaccination against Pseudomonas aeruginosa in patients with cystic fibrosis. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised trials (published or unpublished) comparing Pseudomonas aeruginosa vaccines (oral, parenteral or intranasal) with control vaccines or no intervention in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: We planned to assess the following outcomes: time to infection with Pseudomonas aeruginosa, pulmonary function, body mass index, Schwachman score, frequency of pulmonary infective exacerbations, days of antibiotic usage, days unable to carry out normal daily activities, adverse events, mortality, antibody levels to Pseudomonas aeruginosa and T cell proliferation and cytokine production in response to Pseudomonas aeruginosa. MAIN RESULTS: One trial which included 17 vaccinated patients, with follow up reported to 10 years met the inclusion criteria. Finding only a single trial, and the lack of information on our predefined outcomes limited analysis. REVIEWER'S CONCLUSIONS: There is a paucity of randomised controlled trials assessing the effectiveness of vaccination against Pseudomonas aeruginosa in cystic fibrosis. Increased understanding of modulation of the immune response by vaccination has led to the development of alternative vaccines. We suggest that there is an urgent need for newer vaccines to be evaluated in adequately-powered, multicentre randomised controlled trials examining clinically relevant end-points in addition to immunological variables. Such a trial should assess effectiveness over several years, and include follow-up of vaccinees who become colonised with Pseudomonas aeruginosa. PMID- 10796781 TI - Chest physiotherapy compared to no chest physiotherapy for cystic fibrosis. AB - BACKGROUND: Chest physiotherapy is widely used in patients with cystic fibrosis in order to clear mucus from the airways. OBJECTIVES: To determine the effectiveness and acceptability of chest physiotherapy compared to no treatment or spontaneous cough alone to improve mucus clearance in cystic fibrosis. SEARCH STRATEGY: Relevant trials are identified in the Cochrane Cystic Fibrosis and Genetic Disorders Group Specialised Register of Controlled Trials. This register was compiled by conducting computerised searches of Medline from 1966 to present and from Embase from 1974 to 1995. The register of randomised controlled trials is updated every three months. Unpublished work has been identified by searching through the abstract books of the three major cystic fibrosis conferences; the International Cystic Fibrosis Conference: the European Cystic Fibrosis Conference and the North American Cystic Fibrosis Conference. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials in which a form of chest physiotherapy (airway clearance technique) were taken for consideration in patients with cystic fibrosis compared with either no physiotherapy treatment or spontaneous cough alone. DATA COLLECTION AND ANALYSIS: There were no randomised controlled trials or cross over trials eligible for inclusion in the review. MAIN RESULTS: There were no randomised controlled trials or cross over trials eligible for inclusion in the review. REVIEWER'S CONCLUSIONS: Short-term crossover trials, which had to be excluded from this review, suggest that airway clearance regimens could have beneficial effects in patients with cystic fibrosis. However based on this review there is currently no robust scientific evidence to support the hypothesis that chest physiotherapy for the purpose of clearing airway secretions has a beneficial effect in patients with cystic fibrosis. PMID- 10796782 TI - Newborn screening for cystic fibrosis. AB - BACKGROUND: This review was performed to test the hypothesis that presymptomatic diagnosis, for example by newborn screening, and early treatment may prevent or reduce irreversible organ damage and thereby improve outcome and quality of life in patients with cystic fibrosis. OBJECTIVES: To determine whether there is evidence that early diagnosis of cystic fibrosis by means of neonatal screening, followed by current treatment, improves survival and long term morbidity, without unacceptable adverse effects. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Trials Register. Additional studies were identified by one of the reviewers from handsearching conference proceedings not included in the Cochrane Register. Pharmaceutical companies manufacturing screening tests for cystic fibrosis were also contacted to identify any trials of neonatal screening for cystic fibrosis. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised controlled trials, published and unpublished, comparing screening followed by early treatment to clinical diagnosis and later treatment in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Four reviewers independently assessed trial eligibility and methodological quality and two of these reviewers independently extracted data. MAIN RESULTS: Two trials involving a total of 1,124,483 neonates met inclusion criteria. A total of 210 patients with cystic fibrosis aged from zero to 11 years with a maximum follow-up of eleven years are included. Concealment of allocation was unclear in both studies. Sequence generation was adequate in one study and unclear in the other. Method to ascertain cases was similar in one study and not similar in the other. An intention-to-screen-analysis was possible in one study, but could not be made due to lack of data and was not performed in the other. Differences in study design, variation in outcomes reported and their summary measures precluded calculation of pooled screening estimates. Only data from one study could be analysed in this review. This study reported a reduced risk of weight and height below the fifth percentile among screened patients (odds ratio control compared with screened group for: weight 6.16, 95% Confidence Interval (CI) 2.44, 15.57 and height 5.03, 95% CI 1. 63, 15.63). Adverse effects among parents in the screened and control populations were examined, but it is difficult to assess how meaningful these results are as the timing of the administration of the questionnaire to each group was not clear. Estimation of direct medical costs of screening suggested it was cheaper to diagnose cystic fibrosis by screening rather than other methods. The costing methods used however were not fully described and costs have not been related to effect. REVIEWER'S CONCLUSIONS: There are few randomised controlled trials assessing the effectiveness of neonatal screening in cystic fibrosis. From the data available at this time, there is little evidence suggesting benefit from screening for cystic fibrosis in the neonatal period, although there is similarly little evidence of harm. This systematic review has identified the need for individual patient data from both included studies. Although we have not been able to perform a meta-analysis, this review provides a summary of all the information currently available from randomised controlled trials on the effectiveness of neonatal screening for cystic fibrosis. PMID- 10796783 TI - Absorbent products for the containment of urinary and/or faecal incontinence in adults. AB - BACKGROUND: Incontinence is a distressing condition with significant medical, social and economic implications. People suffering from incontinence, who cannot be successfully cured, depend, almost exclusively, on the use of containment products to manage their symptoms. OBJECTIVES: Many people with incontinence cannot be cured and so depend on symptomatic management. The objective was to assess the effects of different types of absorbent product for the containment of urinary and/or faecal incontinence. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register (March 1999), Embase (to January 1999), Cinahl (to January 1999), HealthSTAR (to January 1999) and the reference lists of relevant articles. We contacted investigators in the field to locate studies. Date of the most recent searches: March 1999. SELECTION CRITERIA: Types of studies All randomised or quasi-randomised trials of absorbent products for the containment of urinary and/or faecal incontinence. Types of participants All adults with urinary and/or faecal incontinence were eligible. The intention was to subdivide participants by severity of underlying incontinence, level of mobility and gender, but this proved not to be feasible. Types of intervention Absorbent products (bodyworns, underpads, and different fabric types for disposable products), for any severity of incontinence. DATA COLLECTION AND ANALYSIS: Trials were evaluated for subject relevance and methodological quality using a standard methodological quality assessment form. If applicable, data on relevant outcomes were then abstracted using a standardised data abstraction form. MAIN RESULTS: Five studies with a total of 345 participants met the selection criteria. Two studies compared disposable with non-disposable bodyworns, one disposable with non-disposable underpads, two fluff pulp with superabsorbent polymers, and one bodyworns with underpads. Data presented on effects were available for few outcomes and were subject to potential bias. REVIEWER'S CONCLUSIONS: The data were too few and of insufficient quality to provide a firm basis for practice. Disposable products may be more effective than non-disposable products in decreasing the incidence of skin problems and superabsorbent products may perform better than fluff pulp products. However, based on the available evidence, these conclusions can only be tentative. PMID- 10796784 TI - Botulinum toxin type A in the treatment of lower limb spasticity in cerebral palsy. AB - OBJECTIVES: To determine whether botulinum toxin (BtA) is an effective and safe treatment for lower limb spasticity in children with cerebral palsy. Functional outcomes are of particular interest. SEARCH STRATEGY: Studies for inclusion in the review were identified using the Movement Disorders Review Group trials register, the Cochrane Controlled Trials Register, MEDLINE, pharmaceutical company databases, communication with other researchers in the field and reference lists of papers found using above search strategies. SELECTION CRITERIA: Studies were considered eligible for inclusion in the review if they evaluated the efficacy of BtA for the treatment of leg spasticity in children with cerebral palsy. They must have been randomised and include a concurrent control group receiving another intervention. DATA COLLECTION AND ANALYSIS: A paper pro forma was used to collect data from the included studies using double extraction by two independent reviewers. Each trial was assessed for internal validity by each of the two reviewers. Meta-analysis was not possible because results were presented in an incompatable form. A Peto odds ratio was calculated where this was appropriate, otherwise a descriptive summary of the results of the individual studies was compiled. MAIN RESULTS: Three eligible studies were found each with small numbers of subjects. They were short term, used single injection sessions with follow-up of between 4 and 26 weeks. One study (Koman), of twelve ambulant children, compared BtA with injection of a placebo and found non significant improvements in gait in the BtA group compared to the placebo group. Two studies (Corry, Flett) compared BtA with the use of casts. Each included 20 ambulant children and found improvements in gait, range of ankle movement and muscle tone in both the BtA and cast groups. However there were no significant differences between the groups in either trial. One of these trials (Flett) also assessed motor function using the gross motor function measure (GMFM) (Russell, 1989) and found significant improvements in each group compared to baseline but no significant differences between the groups. The other trial (Corry) performed 3D gait analysis on those children able to co-operate. Maximal plantar flexion and maximal dorsiflexion during walking were both found to be significantly greater in the BtA group compared to the cast group. In all other dimensions there were no significant differences between the groups. REVIEWER'S CONCLUSIONS: This systematic review has not revealed strong controlled evidence to support or refute the use of BtA for the treatment of leg spasticity in cerebral palsy. Ongoing randomised controlled trials are likely to provide useful data on the short term effects of BtA for leg spasticity. Future research should also assess the longer term use of BtA. Ideally studies should be pragmatic in their approach to dose and distribution of toxin to reflect practise. Outcome measures assessing function and disability would give the most useful information. PMID- 10796785 TI - Gabapentin for drug-resistant partial epilepsy. AB - BACKGROUND: The majority of epileptic patients have a good prognosis and their seizures can be well controlled with the use of a single antiepileptic agent, but up to 30% develop refractory epilepsy, especially those with partial seizures. In this review we summarise the current evidence regarding a new antiepileptic drug, gabapentin, when used as an add-on treatment for drug-resistant partial epilepsy OBJECTIVES: To evaluate the efficacy and tolerability of gabapentin when used as an add-on treatment for patients with drug-resistant partial epilepsy. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group trial register, the Cochrane Controlled Trials Register of The Cochrane Library Issue 4, 1998. In addition, we contacted Parke Davis (manufacturers of gabapentin) and experts in the field to seek any ongoing studies or unpublished studies. SELECTION CRITERIA: Randomized placebo controlled double blind add-on trials of gabapentin in patients with drug resistant partial epilepsy. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted the relevant data. The following outcomes were assessed: (a) 50% or greater reduction in seizure frequency; (b) treatment withdrawal (any reason); (c) side effects. Primary analyses were intention to treat. Sensitivity best and worst case analyses were also undertaken. Summary odds ratios were estimated for each outcome. Dose response was evaluated in regression models, and Number Needed to Treat (NNTs) were calculated for individual doses. MAIN RESULTS: Four trials were included representing 750 randomized patients. EFFICACY: Overall odds ratio (OR) (95% Confidence Interval (CI)) for 50% or greater reduction in seizure frequency compared to placebo 2.22 (1.49 - 3.32). Dose regression analysis shows increasing efficacy with increasing dose, with 28.5% (21.5 - 36.7) of patients responding to 1800mg of gabapentin compared to placebo, NNT 6.7 (3.0 - 10.5). Global effectiveness: treatment withdrawal OR (95% CI) compared to placebo 1.40 (0.76 - 2.55); Side effects: OR (99% CI) compared to placebo. dizziness 2.26 (1.28 - 3.99); fatigue 2.29 (1.11 - 4.74); somnolence 2.04 (1.21 - 3.45) were significanty associated with gabapentin. REVIEWER'S CONCLUSIONS: Gabapentin has efficacy as an add-on treatment in patients with drug-resistant partial epilepsy. However, trials reviewed were of relatively short duration, and provide no evidence for the long term efficacy of gabapentin. Results cannot be extrapolated to monotherapy or patients with other epilepsy types. PMID- 10796786 TI - Zonisamide for drug-resistant partial epilepsy. AB - BACKGROUND: The majority of epileptic patients have a good prognosis and their seizures can be well controlled with the use of a single antiepileptic agent, but up to 30% develop refractory epilepsy, especially those with partial seizures. In this review we summarise the current evidence regarding a new antiepileptic drug, zonisamide, when used as an add-on treatment for drug-resistant partial epilepsy. OBJECTIVES: To evaluate the efficacy and tolerability of zonisamide when used as an add-on treatment for patients with drug -resistant partial epilepsy. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group trial register, the Cochrane Controlled Trials Register (Cochrane Library Issue 4, 1999). In addition, we contacted Dianippon and Elan Pharma (makers and licensees of zonisamide) and experts in the field to seek any ongoing studies or unpublished studies. SELECTION CRITERIA: Randomized placebo controlled add-on trials of zonisamide in patients with drug-resistant partial epilepsy. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted relevant data. The following outcomes were assessed: (a) 50% or greater reduction in total seizure frequency; (b) treatment withdrawal (any reason); (c) adverse events. Primary analyses were intention to treat. Sensitivity best and worst case analyses were also undertaken. Summary odds ratios (ORs) were estimated for each outcome. MAIN RESULTS: Three trials were included with 499 patients randomized. EFFICACY: Overall odds ratio (OR, 95% CI) for 50% reduction in seizure frequency compared to placebo was 2.07 (1.36,3.15) for a 400mg/day dose of zonisamide. When the full treatment period of 12 weeks was considered for all three trials including varied rates of titration to 400mg/day the OR compared to placebo was 2.72 (1.74,4. 25). There was insufficient evidence to support a dose response relationship between dose and responder rate. Tolerability: Treatment withdrawal OR compared to placebo was 1.74 (1.03,2.95). Adverse events: ataxia 3.94 (1.23,12.57), somnolence 2.11 (1.11, 3. 98), agitation 3.52 (1.26,9.68), agitation and irritability 2.43(1. 04,5.66) and anorexia 2.98(1.38,6.42) were significantly associated with zonisamide. REVIEWER'S CONCLUSIONS: Zonisamide has efficacy as an add-on treatment in patients with drug-resistant partial epilepsy. Minimum effective and maximum tolerated doses cannot be identified. The trials reviewed were of 12 week duration and results cannot be used to conform longer periods of effectiveness in seizure control. The results cannot be extrapolated to monotherapy or to patients with other seizure types or epilepsy syndromes. PMID- 10796787 TI - Topiramate for drug-resistant partial epilepsy. AB - BACKGROUND: The majority of epileptic patients have a good prognosis and their seizures can be well controlled with the use of a single antiepileptic agent, but up to 30% develop refractory epilepsy, especially those with partial seizures. In this review we summarize the current evidence regarding a new antiepileptic drug, topiramate, when used as an add-on treatment for drug-resistant partial epilepsy. OBJECTIVES: To evaluate the efficacy and tolerability of topiramate when used as an add-on treatment in patients with drug resistant partial epilepsy. SEARCH STRATEGY: (a) The Cochrane Library (1999 Issue 1); (b) The controlled trial register of the Cochrane Epilepsy Group; (c) Johnson and Johnson, makers of topiramate; (d) Experts in the field. SELECTION CRITERIA: Randomized placebo controlled add-on trials of topiramate in patients with drug resistant epilepsy. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted the relevant data. The following outcomes were assessed: (a) 50% or greater reduction in seizure frequency; (b) treatment withdrawal (any reason); (c) side effects. Primary analyses were intention to treat. Summary odds ratios (OR) were estimated for each outcome. Dose response was evaluated in regression models. MAIN RESULTS: Six trials were included representing 743 randomized patients. EFFICACY: Overall OR (95% CIs) for 50% or greater reduction in seizure frequency compared to placebo 4.06 (2.86-5.78). Dose regression analysis shows increasing efficacy with increasing dose, but found no advantage for doses over 400 mg per day. Global effectiveness: treatment withdrawal OR (95% CIs) compared to placebo 2.57 (1.65-4.00). Side effects: OR (99% CIs)compared to placebo, dizziness 1.99 (1.20-3.29); fatigue 2.52 (1. 47-4.32); nausea 2.84 (1.36 5.93); somnolence 2.89 (1.72-4.85) and 'thinking abnormally' 3.71 (2.02-6.80) were significantly associated with topiramate. REVIEWER'S CONCLUSIONS: Topiramate has efficacy as an add-on treatment in patients with drug resistant partial epilepsy. However, trials reviewed were of relatively short duration, and provide no evidence for the long term efficacy of topiramate. Results cannot be extrapolated to monotherapy or patients with other epilepsy types. PMID- 10796788 TI - Chemotherapy for advanced ovarian cancer. Advanced Ovarian Cancer Trialists Group. AB - BACKGROUND: Ovarian carcinoma is the seventh most common cancer of women in the world and is responsible for the greatest number of deaths from gynaecological malignancy in Europe and North America. Although many studies have explored the use of chemotherapy in this disease, most individual trials have been too small to show clear benefit of one type of chemotherapy over another. OBJECTIVES: The type and intensity of chemotherapy used routinely for women with advanced ovarian cancer has varied because of uncertainty about the effectiveness of the different regimens. The objective of this review was to compare single drugs versus combinations of drugs, platinum versus non-platinum, and carboplatin versus cisplatin-based chemotherapy in women with advanced ovarian cancer. SEARCH STRATEGY: Search strategy: We searched MEDLINE, and CancerLit bibliographic databases and the National Cancer Institute and the UK Co-ordinating Committee on Cancer Research registers of trials. We also handsearched the proceedings of meetings and contacted experts in the field and drug companies. SELECTION CRITERIA: Randomised trials of: (1) single non-platinum versus non-platinum combination chemotherapy (2) single non-platinum versus platinum combination chemotherapy (3) non-platinum regimen versus the same regimen plus cisplatin (4) single platinum versus platinum combination chemotherapy (5) cisplatin versus carboplatin-based chemotherapy in women with advanced ovarian cancer. DATA COLLECTION AND ANALYSIS: Individual patient data were obtained from the trial investigators, checked by the reviewers and finally verified by the trial investigator. MAIN RESULTS: Main results: 49 trials involving 8763 women were included. The data were combined to calculate hazard ratios (HR) for survival on an intention-to-treat basis. For single non-platinum versus platinum combination chemotherapy the overall HR for survival was 0.93, 95% confidence interval (CI) 0.83-1.05 in favour of platinum-based combination chemotherapy. For non-platinum regimens compared with the same regimen plus cisplatin the survival HR was 0.88, 95% CI 0.79- 0.98 in favour of the addition of platinum to drug regimens. Single platinum compared with platinum combination therapy gave a HR of 0.91, 95% CI 0.79-1.05 in favour of combination chemotherapy. Cisplatin versus carboplatin gave a HR of 1.02, 95% CI 0.93-1.12. Sub-group analyses for age, stage, grade, histology, resection, bulk of residual tumour and performance status were undertaken for cisplatin versus carboplatin only. No difference in effect was found. REVIEWER'S CONCLUSIONS: The available evidence, although not conclusive, suggests that platinum-based chemotherapy is better than non-platinum therapy. There is some evidence that combination therapy improves survival compared with platinum alone. No difference in effect has been shown between cisplatin and carboplatin. PMID- 10796789 TI - Interventions for preventing blood loss during the treatment of cervical intraepithelial neoplasia. AB - OBJECTIVES: Good surgical technique and other interventions can reduce immediate and long-term blood loss which is a common complication of surgery for cervical intra-epithelial neoplasia. The objective of this review was to assess the effect of interventions to reduce blood loss due to surgery for cervical intra epithelial neoplasia. SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Group trials register. SELECTION CRITERIA: Randomised and quasi-randomised trials of vasopressin, tranexamic acid, haemostatic sutures or Monsel's solution in women who have had surgery for intra-epithelial neoplasia. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted data. MAIN RESULTS: Seven trials were included. Vasopressin had a beneficial effect on peri-operative bleeding. Prophylactic tranexamic acid reduced haemorrhage after knife and laser cone biopsy. Packing with Monsel's solution resulted in less peri-operative blood loss and secondary haemorrhages than routine suturing. Elective suturing also appeared to increase the risk of amenorrhoea and dysmenorrhoea. Vaginal packing increased satisfactory colposcopy rates at follow-up and reduced post-surgical stenosis. REVIEWER'S CONCLUSIONS: Bleeding associated with surgery of the cervix appears to be reduced by vasopressin, which is used under local rather than general anaesthetic. Tranexamic acid appears to be beneficial after knife and laser cone biopsy. There are not enough data to assess the effects on primary haemorrhage. There is some evidence that haemostatic suturing has an adverse effect on blood loss, cervical stenosis and satisfactory colposcopy. PMID- 10796790 TI - Serenoa repens for benign prostatic hyperplasia. AB - OBJECTIVES: This systematic review aimed to assess the effects of Serenoa repens in the treatment of Benign Prostatic Hyperplasia (BPH). SEARCH STRATEGY: Trials were searched in computerized general and specialized databases (MEDLINE, EMBASE, Cochrane Library, Phytodok), by checking bibliographies, and by contacting manufacturers and researchers. SELECTION CRITERIA: Trials were eligible if they (1) randomized men with BPH to receive preparations of Serenoa repens (alone or in combination) in comparison with placebo or other BPH medications, and (2) included clinical outcomes such as urologic symptom scales, symptoms, or urodynamic measurements. Eligibility was assessed by at least two independent observers. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes was extracted by at least two independent reviewers using a standard form. The main outcome measure for comparing the effectiveness of Serenoa repens with placebo or other BPH medications was the change in urologic symptom scale scores. Secondary outcomes included changes in nocturia and urodynamic measures. The main outcome measure for side effects was the number of men reporting side effects. MAIN RESULTS: 2939 men from18 randomized trials lasting 4 to 48 weeks were assessed. 16 trials were double-blinded and treatment allocation concealment was adequate in 9 studies. Compared with placebo, Serenoa repens improved urinary symptom scores, symptoms, and urinary flow measures. The weighted mean difference (WMD) for the urinary symptom score was -1.41 points (scale range 0-19), (95%CI = -2.52, -0.30, n = 1 study) and the risk ratio (RR) for self rated improvement was 1.75 (95%CI = 1.21, 2.54, n = 6 studies). The WMD for nocturia was -0.76 times per evening (95%CI = -1.22, -0.32; n = 10 studies). The WMD for peak urine flow was 1.93 ml/sec (95%CI = 0.72, 3.14, n = 8 studies). Compared with finasteride, Serenoa repens produced similar improvements in urinary symptom scores (WMD = 0.37 IPSS points (scale range 0-35), 95%CI = -0.45, 1.19, n = 2 studies) and peak urine flow (WMD = -0.74 ml/sec, 95%CI = -1.66, 0.18, n = 2 studies). Adverse effects due to Serenoa repens were mild and infrequent. Withdrawal rates in men assigned to placebo, Serenoa repens or finasteride were 7%, 9%, and 11%, respectively. REVIEWER'S CONCLUSIONS: The evidence suggests that Serenoa repens improves urologic symptoms and flow measures compared with placebo. Serenoa repens produced similar improvement in urinary symptoms and flow compared to finasteride and is associated with fewer adverse treatment events. The long term effectiveness, safety and ability to prevent BPH complications are not known. PMID- 10796791 TI - Neoadjuvant cisplatin for advanced bladder cancer. AB - OBJECTIVES: The main objective of this review was to investigate whether platinum based chemotherapy given either before or during local treatment, improves the survival of patients with locally advanced bladder cancer. A further objective was to determine whether there is any evidence that such chemotherapy is more or less effective within well defined subgroups of patients. SEARCH STRATEGY: MEDLINE and CANCERLIT bibliographic searches were supplemented by information obtained from trial registers and, by hand searching relevant meeting proceedings, and by discussion with relevant trialists and organisations. SELECTION CRITERIA: Trials were included in the meta-analysis provided they were properly randomised, included patients with advanced bladder cancer and compared local treatment versus the same local treatment plus neoadjuvant or concurrent chemotherapy. DATA COLLECTION AND ANALYSIS: Updated individual patient data were sought from the trialists responsible for all eligible randomised controlled trials (all were unpublished at outset of meta-analysis). Time to event analyses of survival were done on intention to treat basis. A sensitivity analysis including summary data from the single trial for which individual patient data were not available was also done. Pre-defined subgroup analyses by age, sex, tumour stage and grade were also carried out. MAIN RESULTS: Individual data on 479 patients from 4 randomised trials were available. Data extracted from a published report was used for 1 further trial (325 patients) in a supplementary analysis. Analysis of the individual patient data gave an overall hazard ratio of 1.02 in favour of local therapy alone (P=0.845, 95% confidence interval=0.81 1.26). When this analysis was supplemented by data from the only trial for which individual patient information was not available, the hazard ratio was 0.91 in favour of chemotherapy (P=0.328, 95% confidence interval=0.75-1.10). Neither analysis was conventionally significant. The only prognostic factor for which the evidence suggested a differential treatment effect (interaction) across groups was age (chi-square test for trend=3.833, P=0.05), with younger age groups (<60 years) showing a possible effect in favour or chemotherapy. REVIEWER'S CONCLUSIONS: There is insufficient information currently included in this meta analysis to obtain a definitive answer to the question of whether neoadjuvant cisplatin-based chemotherapy improves the survival of patients with locally advanced bladder cancer. Since the publication of this review by the collaborative group in 1995, 4 additional trials have been completed, although none of these has yet been published in full. The next update of the meta analysis (planned for 1999/2000) will aim to include source data from these trials and should therefore provide more definitive results. PMID- 10796792 TI - Topical treatments for fungal infections of the skin and nails of the foot. AB - OBJECTIVES: The objective of this review is to identify and evaluate the evidence of efficacy for topical treatments for fungal infections of the skin and nails of the human foot. To establish the effectiveness of topical treatments in achieving a cured condition and in preventing recurrence SEARCH STRATEGY: Randomised controlled trials were identified from the MEDLINE, EMBASE and CINHAL databases, from the beginning of these databases to December 1997. Also we screened the Cochrane Controlled Trials Register, the Science Citation Index BIOSIS, CAB - Health, Healthstar and Economic databases. References and unpublished studies were searched, podiatry journals handsearched and the pharmaceutical industry contacted. SELECTION CRITERIA: Only randomised controlled trials (RCTs) using participants who have mycologically diagnosed fungal infections of the skin and nails of the human foot are included in the analysis. DATA COLLECTION AND ANALYSIS: Two reviewers independently summarised the included trials and appraised their quality of reporting using a structured data extraction tool which assessed 12 quality criteria. MAIN RESULTS: Of 126 trials identified in 121 papers, 72 met the inclusion criteria. Placebo-controlled trials yielded the following pooled relative risks of failure to cure (RRFC) for skin infections: allylamines 0.30 (95% confidence interval 0.23 to 0.37); azoles 0.53 (0.42 to 0.68); undecenoic acid 0.28 (0.11 to 0.74); tolnaftate 0.46 (0.17 to 1.22). Though meta-analysis of 11 trials comparing allylamines and azoles showed an RRFC of 0.88 (0.78 to 0.99) in favour of allylamines, there was evidence of language bias. Seven English language reports favoured allylamines (RRFC = 0.79; 0.68 to 0.93) but four foreign language reports showed no difference between the two drugs (RRFC = 1.00; 0.90 to 1.12). The two trials of nail infections did not provide any evidence of benefit for topical treatments compared with placebo. REVIEWER'S CONCLUSIONS: CONCLUSIONS: In placebo-controlled trials allylamines, azoles and undecenoic acid were efficacious. There are sufficient comparative trials to judge relative efficacy only between allylamines and azoles. Allylamines cure slightly more infections than azoles but are much more expensive. The most cost-effective strategy is first to treat with azoles or undecenoic acid and to use allylamines only if that fails. PMID- 10796793 TI - Single dose dextropropoxyphene, alone and with paracetamol (acetaminophen), for postoperative pain. AB - BACKGROUND: Patient surveys have shown that postoperative pain is often not managed well, and there is a need to assess the efficacy and safety of commonly used analgesics as newer treatments become available. Dextropropoxyphene is one example of an opioid analgesic in current use, and is widely prescribed for pain relief in combination with paracetamol under names such as Co-proxamol and Distalgesic. OBJECTIVES: To determine the analgesic efficacy and adverse effects of single dose oral Dextropropoxyphene alone and in combination with paracetamol (acetaminophen) for moderate to severe postoperative pain. SEARCH STRATEGY: Published reports were identified from: Medline (1966 - November 1996), Biological Abstracts (1985 - 1996), Embase (1980 - 1996), the Cochrane Library (Issue 4 1996), and the Oxford Pain Relief Database (1954 - 1994). Additional studies were identified from the reference lists of retrieved reports. Date of the most recent searches: July 1998. SELECTION CRITERIA: The inclusion criteria used were: full journal publication, postoperative pain, postoperative oral administration, adult patients, baseline pain of moderate to severe intensity, double-blind design, and random allocation to treatment groups which included dextropropoxyphene and placebo or a combination of dextropropoxyphene plus paracetamol and placebo. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers, and trials were quality scored. Summed pain intensity and pain relief data were extracted and converted into dichotomous information to yield the number of patients with at least 50% pain relief. This was used to calculate the relative benefit and number-needed-to-treat (NNT) for one patient to achieve at least 50% pain relief. MAIN RESULTS: Six trials (440 patients) compared dextropropoxyphene with placebo and five (963 patients) compared dextropropoxyphene plus paracetamol 650 mg with placebo. For a single dose of dextropropoxyphene 65 mg in postoperative pain the NNT for at least 50% pain relief was 7.7 (95% confidence interval 4.6 to 22) when compared with placebo over 4-6 hours. For the equivalent dose of dextropropoxyphene combined with paracetamol 650 mg the NNT was 4.4 (3.5 to 5.6) when compared with placebo. These results were compared with those for other analgesics obtained from equivalent systematic reviews. Pooled data showed increased incidence of central nervous system adverse effects for dextropropoxyphene plus paracetamol compared with placebo. REVIEWER'S CONCLUSIONS: The combination of dextropropoxyphene 65 mg with paracetamol 650 mg shows similar efficacy to tramadol 100 mg for single dose studies in postoperative pain but with a lower incidence of adverse effects. The same dose of paracetamol combined with 60 mg codeine appears more effective but, with the slight overlap in the 95% confidence intervals, this conclusion is not robust. Adverse effects of both combinations were similar. Ibuprofen 400 mg has a lower (better) NNT than both dextropropoxyphene 65 mg plus paracetamol 650 mg and tramadol 100 mg. PMID- 10796794 TI - Growth monitoring in children. AB - BACKGROUND: Growth monitoring is widely accepted and strongly supported by health professionals, and is a standard component of community paediatric services throughout the world. We sought to evaluate research evidence of its impact. This requires definition, consideration of the setting, and discussion of the intended effects of this activity. In this review, we define growth monitoring as the regular recording of a child's weight, coupled with some specified remedial actions if the weight is abnormal in some way. Although the causes of growth faltering and the responses to it may be region specific, the process is the same, and we consider here growth monitoring in both the deprived and richer populations of the world. OBJECTIVES: Growth monitoring consists of routine measurements to detect abnormal growth, combined with some action when this is detected. As primary care workers worldwide invest time in this activity, we sought evidence of its benefits and harms. The review objectives are to evaluate the effects of routine growth monitoring on: 1. The child, in relation to preventing death, illness or malnutrition; and referrals for medical care, medical specialist assessment or professional social support follow-up. 2. The mother, in relation to nutritional knowledge, anxiety or reassurance about the child's health, and satisfaction with services. SEARCH STRATEGY: Cochrane Controlled Trials Register; MEDLINE; EMBASE; CINAHL; World Health Organization and World Bank publications; specialists in this area; citations in existing reviews and identified studies. SELECTION CRITERIA: Randomised or quasi randomised trials comparing routine growth monitoring (regular monitoring of growth, plotting on a chart, combined with referral or intervention when growth is abnormal) with no growth monitoring. DATA COLLECTION AND ANALYSIS: Trial quality was assessed, and data abstracted by both reviewers. MAIN RESULTS: Two studies included, both conducted in developing countries. In one, the nutritional status at 30 months in 500 children showed no difference between those allocated to growth monitoring and those not. The other study examined whether counselling improved mothers' knowledge of the growth chart, and reported better test scores at four months. REVIEWER'S CONCLUSIONS: Given the level of investment in growth monitoring worldwide, it is surprising there is so little research evaluating its potential benefits and harms. PMID- 10796795 TI - Corticosteroids for treating Guillain-Barre syndrome. AB - BACKGROUND: The cause of Guillain-Barre syndrome (GBS) is inflammation of the peripheral nerves which corticosteroids would be expected to benefit. OBJECTIVES: To examine the efficacy of corticosteroids in hastening recovery and reducing the long term morbidity from Guillain-Barre syndrome (GBS). SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group register for randomised trials and enquiry from authors of trials and other experts in the field. SELECTION CRITERIA: Types of studies: quasi-randomised or randomised controlled trials TYPES OF PARTICIPANTS: patients with GBS of all ages and all degrees of severity Types of interventions: any form of corticosteroid or adrenocorticotrophic hormone Types of outcome measures: Primary: improvement in disability grade on a commonly used seven point scale four weeks after randomisation Secondary: time from randomisation until recovery of unaided walking, time from randomisation until discontinuation of ventilation (for those ventilated), mortality, proportion of patients dead or disabled (unable to walk without aid) after 12 months, improvement in disability grade after six months, improvement in disability grade after 12 months, proportion of patients who relapse, and proportion of patients with adverse events related to corticosteroid treatment DATA COLLECTION AND ANALYSIS: We identified six randomised trials. One author extracted the data and the other checked them. We obtained some missing data from investigators. MAIN RESULTS: The six eligible trials included a total of 195 corticosteroid treated patients and 187 control subjects. One trial of intravenous methylprednisolone accounted for 243 of the total 382 subjects studied (63%). This trial did not show a significant difference in any disability related outcome between the corticosteroid and placebo groups. There was no significant difference between the corticosteroid and control groups for the primary outcome measure, improvement in disability grade four weeks after randomisation. The weighted mean difference of the three trials for which this outcome was available showed no difference. The actual figure was 0.01 (95% CI 0.27 to 0.29) grade in favour of the corticosteroid group. There was also no significant difference between the groups for most of the secondary outcome measures. However in the largest trial hypertension developed less often in the intravenous methylprednisolone group (2/124, 1.6%) than in the control group (12/118, 10.2%), a significant difference in favour of corticosteroid treatment (relative risk 0.20, 95% CI 0.04 to 0.66). REVIEWER'S CONCLUSIONS: Corticosteroids should not be used in the treatment of Guillain-Barre syndrome. If a patient with Guillain-Barre syndrome needs corticosteroid treatment for some other reason its use will probably not do harm. The effect of intravenous methylprednisolone combined with intravenous immunoglobulin in Guillain-Barre syndrome is being tested with a randomised trial. PMID- 10796796 TI - Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND). AB - BACKGROUND: Riluzole has been approved for treatment of patients with amyotrophic lateral sclerosis (ALS) in some countries but not others. Questions persist about its clinical utility because of high cost, modest efficacy and concern over adverse effects. OBJECTIVES: To examine the efficacy of riluzole in prolonging survival, and in delaying the use of surrogates (tracheostomy and mechanical ventilation) to sustain survival. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group Register for randomized trials and enquiry from authors of trials and other experts in the field. The most recent search was conducted in June 1999. SELECTION CRITERIA: Types of studies: randomized trials TYPES OF PARTICIPANTS: adults with a diagnosis of ALS Types of interventions: treatment with riluzole or placebo Types of outcome measures: Primary: per cent mortality at 12 months with riluzole 100 mg Secondary: per cent mortality as a function of time with 100 mg and with all doses of riluzole, scales of neurologic function, quality of life, muscle strength and adverse events. DATA COLLECTION AND ANALYSIS: We identified two randomized trials. Each reviewer graded them for methodological quality. Data extraction was performed by a single reviewer and checked by the other two. We obtained some missing data from investigators. We performed meta-analyses with RevMan software using a fixed effects model. MAIN RESULTS: The two eligible trials included a total of 794 riluzole treated patients and 320 placebo treated patients. The methodological quality was acceptable and the trials were easily comparable. There were significant differences between the riluzole and placebo groups of both trials, in terms of the primary outcome measure, which was per cent mortality at 12 months with the 100 mg dose of riluzole. The odds ratio for the combined studies was 0.57 (95%CI 0.41 to 0.80) at 12 months. In the secondary outcome measures, there was a survival advantage with riluzole 100 mg at six, nine, 12 and 15 months, but not at three or 18 months. Pooled data from the 50, 100 and 200mg dose groups in the larger trial showed a lower per cent mortality with riluzole compared to placebo only at 12 months (odds ratio (OR) 0.64, 95% CI 0.47 to 0.88). There was no beneficial effect on bulbar function, or muscle strength. There were scant data on quality of life, but patients treated with riluzole remained in a more moderately affected health state significantly longer than placebo-treated patients (weighted mean difference (WMD) 35.5 days, 95% CI 5.9 to 65. 0). A threefold increase in serum alanine transferase was more frequent in riluzole treated patients than controls (WMD 2.65, 95% CI 1.51 to 4.65). REVIEWER'S CONCLUSIONS: Riluzole 100 mg per day appears to be modestly effective in prolonging survival for patients with ALS. PMID- 10796797 TI - Oral non-steroidal anti-inflammatory drug therapy for cystic fibrosis. AB - BACKGROUND: Maintenance of optimal lung function is an important therapeutic goal in cystic fibrosis as it is lung damage that, in the long term, is responsible for most premature death among affected people. It has been hypothesised that lung damage results from inflammation and that prolonged use of non-steroidal anti-inflammatory drugs may prevent progressive pulmonary deterioration and respiratory morbidity in cystic fibrosis. It is thus important to establish the current level of evidence about the potential benefits and harms of treatment with non-steroidal anti-inflammatory drugs. OBJECTIVES: The aim of this systematic review is to assess the effectiveness of treatment with non-steroidal anti-inflammatory agents in cystic fibrosis. SEARCH STRATEGY: Trials were ascertained from the Cochrane Cystic Fibrosis and Genetic Disorders Specialised Register of Controlled Trials which includes published and unpublished trials identified through electronic databases such as Medline and Embase as well as those identified from handsearching of journals and conference proceedings. Pharmaceutical companies manufacturing non-steroidal anti-inflammatory drugs were also contacted to identify any trials of non-steroidal anti-inflammatory drugs in cystic fibrosis. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised controlled trials, published and unpublished, comparing non-steroidal anti inflammatory drugs, administered orally at any dose for a period of at least two months, to placebo in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: The following outcomes were assessed: objective measures of lung function, nutritional status, radiological assessment of pulmonary involvement, use of intravenous antibiotics, hospital admissions, survival, frequency of major and minor adverse effects and compliance with therapy. MAIN RESULTS: Three trials involving 145 patients aged from five to 39 years with a maximum follow up of four years met the inclusion criteria. Methodological quality was deemed good or adequate in two. Two trials, both reporting effectiveness of ibuprofen in subjects with mild lung disease, were from the same centre and included some patients in common, while the third assessed piroxicam in subjects with more severe impairment of respiratory function. Variation in outcomes reported and their summary measures precluded calculation of pooled treatment estimates. Only one trial reported within-subject changes in pulmonary function and the findings of this trial suggested that there was a greater absolute annual decline in percentage predicted forced expiratory volume in one second among controls than among those treated with ibuprofen. In a post-hoc sub-group analysis this effect was confined to children aged five to 13 years. In addition, in this one trial long term use of high dose ibuprofen was associated with reduced intravenous antibiotic usage, improved nutritional and radiological pulmonary status. No major adverse effects were reported but the power of the trials to identify clinically important differences in the incidence of adverse effects was low. REVIEWER'S CONCLUSIONS: While there is preliminary evidence to suggest that non steroidal anti-inflammatory drugs may prevent pulmonary deterioration in subjects with mild lung disease due to cystic fibrosis, currently their routine use cannot be recommended. Further trials are required to confirm that their use prevents pulmonary deterioration and is associated with improved nutritional status. Such trials should also address the age group of subjects most likely to benefit, the prevalence of important adverse effects and the optimal dosage schedule as well as any reduction in concomitant therapy. Multi-centre trials will add to the validity of findings by enhancing their generalisability. The question of whether anti-inflammatory treatment prevents lung damage in pre-symptomatic PMID- 10796798 TI - Nebulised hypertonic saline for cystic fibrosis. AB - BACKGROUND: The lung disease in cystic fibrosis is characterised by impaired mucociliary clearance, recurrent bronchial infection and airway inflammation. Hypertonic saline has been shown to enhance mucociliary clearance in-vitro and this may act to lessen the destructive inflammatory process in the airways. OBJECTIVES: To determine if nebulised hypertonic saline treatment improved lung function, exercise tolerance, quality of life and decreased the incidence of exacerbations of respiratory infections in patients with cystic fibrosis. SEARCH STRATEGY: Studies were identified from the Cochrane Cystic Fibrosis and Genetic Disorders Group trials register. Titles and abstracts were reviewed to identify all controlled trials. Review articles and bibliographies identified from this process were surveyed for additional citations & RCTs. Identification of unpublished work was obtained from abstract books from the three major Cystic Fibrosis conferences (International Cystic Fibrosis Conference, The European Cystic Fibrosis Conference and the North American Cystic Fibrosis Conference). Trial authors were contacted for additional information when only abstracts were available to review. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All controlled trials that assessed the effect of hypertonic saline compared to placebo or other mucolytic therapy, for any duration or dose regimen in subjects with cystic fibrosis of any age or severity were reviewed. Studies in languages other than English were included. DATA COLLECTION AND ANALYSIS: All identified trials were independently reviewed by both reviewers & all data collected. Trial quality was scored by the Cochrane assessment of allocation concealment & the Jadad scale of methodological quality. MAIN RESULTS: Twelve controlled trials of hypertonic saline were identified. Seven trials met the inclusion criteria; these involved 143 subjects with an age range of 6 to 46 years. Of these, six were published studies and one in abstract form. The durations of the trials were limited to immediate effects on mucociliary clearance to a maximum of three weeks. In two studies, involving thirty five subjects, a score for the feeling of cleared chest was made using visual analogue scales. This analysis showed a weighted mean difference of -0.98 (95% confidence Interval -1.6, -0.34), favouring hypertonic saline over isotonic saline. In two trials with 22 subjects hypertonic saline improved mucociliary clearance as measured by isotope clearance from the lungs in 90 minutes demonstrating a weighted mean difference of -11.3 (95% confidence Interval -18.6, -4.0), and as area under the clearance time curve; weighted mean difference of 212 (95%CI -272, -152), also favouring hypertonic saline over isotonic saline. Lung function as measured by improvement in FEV1 was observed in one study of 27 subjects. The percentage increase in FEV1 at two weeks increased by a mean 15.0% with hypertonic saline and 2.8% with isotonic saline (p=0.004). Adverse events were adequately described in only one trial and none were serious. REVIEWER'S CONCLUSIONS: Nebulised hypertonic saline improves mucociliary clearance immediately after administration which may have a longer term beneficial effect in cystic fibrosis. The maximum time data were recorded for was only three weeks. Most of the patients had mild to moderate lung disease and the effect on severe lung disease remains unclear. Further studies of hypertonic saline should be carried out to determine the effect on pulmonary function tests, quality of life, frequency of exacerbations of respiratory disease and efficacy comparisons with nebulised deoxyribonuclease, with larger numbers and for longer duration. At this stage there is insufficient evidence to support the use of hypertonic saline in routine treatment for patients with cystic fibrosis. PMID- 10796799 TI - Tyrosine supplementation for phenylketonuria. AB - BACKGROUND: Phenylketonuria is an inherited disease for which the main treatment is the dietary restriction of the amino acid phenylalanine. The diet has to be initiated in the neonatal period to prevent or reduce mental handicap however the diet is very restrictive and unpalatable and can be difficult to follow. A deficiency of the amino acid tyrosine has been suggested as a cause of some of the neuropsychological problems exhibited in PKU. Therefore, this review aims to assess the efficasy of tyrosine supplementation for phenylketonuria. OBJECTIVES: To assess the effects of tyrosine supplementation alongside or instead of phenylalanine restricted diet for patients with phenylketonuria who commenced on diet at diagnosis and either continued on the diet or relaxed the diet later in life. To assess the evidence that tyrosine supplementation alongside, or instead of phenylalanine restricted diet improves intelligence, neuropsychological performance, growth and nutritional status, mortality rate and quality of life. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Trials Register which is a specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Additional studies were identified from handsearching the Journal of Inherited Metabolic Disease (from inception, 1978, to 1998). The manufacturers of prescribable dietary products used in the treatment of phenylketonuria were also contacted for further references. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudo randomised trials investigating the use of tyrosine supplementation versus placebo in patients with phenylketonuria in addition to, or instead of, a phenylalanine restricted diet. Patients treated for maternal phenylketonuria were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the trial eligibility, methodological quality and extracted the data. MAIN RESULTS: Two trials were included with a total of 47 patients. The blood tyrosine concentrations were significantly higher in the patients receiving tyrosine supplements than those in the placebo group (weighted mean difference 22.526, 95% Confidence interval (CI) 12.182 - 32.870). No significant differences were found between any of the other outcomes measured. REVIEWER'S CONCLUSIONS: From the available evidence no recommendations can be made about whether tyrosine supplementation should be introduced into routine clinical practice. Further randomised controlled studies are required to provide further evidence. PMID- 10796800 TI - Lisuride versus bromocriptine for levodopa-induced complications in Parkinson's disease. AB - OBJECTIVES: To compare the efficacy and safety of adjunct lisuride therapy versus bromocriptine in patients with Parkinson's disease, already established on levodopa and suffering the long-term complications of therapy. SEARCH STRATEGY: Electronic searches of MEDLINE, EMBASE and the Cochrane Controlled Trials Register. Handsearching of the neurology literature as part of the Cochrane Movement Disorders Group's strategy. Examination of the reference lists of identified studies and other reviews. Contact with Cambridge Laboratories, Roche Products Limited and Sandoz Limited. SELECTION CRITERIA: Randomised controlled trials of lisuride versus bromocriptine in patients with a clinical diagnosis of idiopathic Parkinson's disease and long-term complications of levodopa therapy. DATA COLLECTION AND ANALYSIS: Data was abstracted independently by each author and differences settled by discussion. MAIN RESULTS: Only one randomised cross over trial including 20 patients has compared lisuride with bromocriptine as adjunct therapy in Parkinson's disease. Both lisuride and bromocriptine improved motor fluctuations with no significant differences between the agonists. However, this conclusion is based on an unvalidated 4 point rating scale which could only record positive outcomes. This, combined with the small size of the trial, suggests that firm conclusions on motor fluctuations should not be drawn. Lisuride and bromocriptine produced similar benefits in parkinsonian impairments according to the Columbia Rating Scale. Adverse events were similar with the two agonists and no withdrawals were reported from either drug. REVIEWER'S CONCLUSIONS: The small size of this study and other methodological problems do not allow any firm conclusions to be drawn regarding the efficacy and safety of lisuride compared with bromocriptine in advanced Parkinson's disease with motor complications. PMID- 10796801 TI - Lisuride for levodopa-induced complications in Parkinson's disease. AB - OBJECTIVES: To compare the efficacy and safety of adjuvant lisuride therapy versus placebo in patients with Parkinson's disease, already established on levodopa and suffering from motor complications. SEARCH STRATEGY: Electronic searches of MEDLINE, EMBASE and the Cochrane Controlled Trials Register. Handsearching of the neurology literature as part of the Cochrane Movement Disorders Group's strategy. Examination of the reference lists of identified studies and other reviews. Contact with Cambridge Laboratories and Roche Products Limited. SELECTION CRITERIA: Randomised controlled trials of lisuride versus placebo in patients with a clinical diagnosis of idiopathic Parkinson's disease and long-term complications of levodopa therapy. DATA COLLECTION AND ANALYSIS: Data was to be abstracted independently by each author and differences settled by discussion. MAIN RESULTS: No randomised controlled trials comparing lisuride with placebo in advanced Parkinson's disease with motor complications were found. REVIEWER'S CONCLUSIONS: Well designed randomised controlled trials demonstrating efficacy and safety are required before the use of lisuride in later Parkinson's disease can be supported. PMID- 10796802 TI - Interventions for pathological gambling. AB - BACKGROUND: With the legalization of new forms of gambling there are increasing numbers of individuals who appear to have gambling related problems and who are seeking help. The individual and societal consequences are significant. Pathological gambling can result in the gambler jeopardizing or losing a significant relationship or job and committing criminal offences. Pathological gamblers may develop general medical conditions associated with stress. Increased rates have been reported for mood disorders, attention-deficit/hyperactivity disorder, substance abuse or dependence. There is a high risk of suicide and a high correlation with antisocial, narcissistic and borderline personality disorders and alcohol addiction. With increasing public awareness of gambling related problems health funders and practitioners are asking questions about the efficacy of treatments. Consequently quality research into gambling treatment is crucial. OBJECTIVES: The objective of this review was to complete a systematic review and meta-analysis of all randomised controlled trials (RCTs) of psychological and pharmacological treatments for pathological gambling, from both published and unpublished scientific reports. SEARCH STRATEGY: Published and unpublished RCTs of treatments of pathological gambling were identified by searches of electronic databases and hand searching journals likely to contain RCTs of gambling treatments. Researchers and gambling treatment centres were contacted by letter. Bibliographies of all identified research studies were scanned to identify other relevant references. SELECTION CRITERIA: All RCTs of treatments for pathological gambling were eligible for inclusion. DATA COLLECTION AND ANALYSIS: The data was entered into the Cochrane Review Manager software (REVMAN). The component RCTs were quality rated, with special emphasis on the concealment of treatment allocation and blinding. Relative risk analyses were conducted for the dichotomous outcome of controlled vs. uncontrolled gambling. The relative risks were aggregated using both fixed and random effects models. Tests for heterogeneity were undertaken. Both short-term (1 month or less) and long-term (6 months or longer) outcomes were considered. MAIN RESULTS: Only four RCTs of psychological treatments were identified. These RCTs were heterogeneous in terms of design, interventions, outcome measurement and follow-up periods. All had small numbers of participants. The studies had poor methodological quality features. The experimental interventions, behavioural or cognitive-behavioural therapy (BT/CBT), were more efficacious than the control interventions in the short-term (relative risk 0.44, 95% confidence interval (CI) 0.24-0. 81). There was a trend for long-term treatment with BT/CBT to be more efficacious than the control treatments, but the statistical significance of this was sensitive to the statistical model used for meta-analysis. With a fixed effect model the relative risk was 0.56 (95% CI 0.33-0.95); the relative risk with a random effects model was 0.61 (95% CI 0.25-1.47). REVIEWER'S CONCLUSIONS: This systematic review revealed a paucity of evidence for effective treatment of pathological gambling. As gambling is becoming more accessible in many countries and there is epidemiological evidence of increasing rates of pathological gambling, more rigorous RCTs are required. PMID- 10796803 TI - Yoga for epilepsy. AB - BACKGROUND: Stress is considered an important precipitating factor for seizures. Yoga is believed to induce relaxation and stress reduction. The effect of yoga on the EEG and the autonomic nervous system have been reported. Yoga would be an attractive therapeutic option for epilepsy (if proved effective), in view of its nonpharmacological nature, minimal side effects and international acceptance. OBJECTIVES: To assess the efficacy of yoga in the treatment of patients with epilepsy. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group trial register, the Cochrane Controlled Trials Register (The Cochrane Library Issue 4, 1998), MEDLINE for articles published up to the middle of 1998, and also registries of the research council for complimentary medicine were searched. In addition, we searched the references of all the identified studies. Finally, we contacted the members of the Neurological Society of India, several neurophysiology institutions and yoga institutes to seek any ongoing studies or studies published in nonindexed journals or unpublished studies. SELECTION CRITERIA: Randomized control trials and controlled clinical trials of treatment of epilepsy with yoga. DATA COLLECTION AND ANALYSIS: The data were extracted independently by both reviewers and any discrepancies were resolved by discussion. The main outcomes assessed were percentage of patients rendered seizure free, number of patients with more than 50% reduction in seizure frequency or seizure duration and the overall reduction in seizure frequency. Analyses were on an intention to treat basis. MAIN RESULTS: Only one study met the selection criteria, and recruited a total of 32 patients, 10 to sahaja yoga and 22 to control treatments. Antiepileptic drugs were continued in all. Randomization was by roll of a dice. The results of this study are as follows: (i) Four patients treated with yoga were seizure free for six months compared to none in the control groups. The Odds Ratio (OR) (95% Confidence Interval (CI)) for yoga versus sham yoga group was 14.5 (0.7, 316.7) and for yoga versus no treatment group 17.3 (0.8, 373.5). (ii) Nine patients in the yoga group had more than 50% reduction in seizure frequency compared to only one among the controls. The OR (95% CI) for yoga versus sham yoga group was 81 (4.4, 1504.5) and for the yoga versus no treatment group was 158.3 (5.8, 4335.9). (iii) There was a decline in the average number of attacks per month compared to the baseline frequency among the patients treated with yoga. The weighted mean difference ( 95% CI) between yoga versus sham yoga group was -2.1 (-3.1, -1.0) and for the yoga versus no treatment group -1.1 (-1.8, -0.4). (iv) More than 50% reduction in seizure duration was found in seven of the 10 patients treated with yoga, compared to none among the 22 controls. The OR (95%CI) for yoga versus sham yoga group was 45 (2.0, 1006.8) and for yoga versus no treatment group 53.57 (2.4, 1187.3). REVIEWER'S CONCLUSIONS: No reliable conclusions can be drawn regarding the efficacy of yoga as a treatment for epilepsy. Further studies are necessary to evaluate the efficacy of yoga in the treatment of epilepsy. PMID- 10796804 TI - Maximal androgen blockade for advanced prostate cancer. AB - OBJECTIVES: This systematic review assessed the effect of maximal androgen blockade (MAB) on survival when compared to castration (medical or surgical) alone for patients with advanced prostate cancer. SEARCH STRATEGY: Randomized controlled trials were searched in general and specialized databases (MEDLINE, EMBASE, Cancerlit, Cochrane Library, VA Cochrane Prostate Disease register) and by reviewing bibliographies. SELECTION CRITERIA: All published randomized trials were eligible for inclusion provided they (1) randomized men with advanced prostate cancer to receive a non-steroidal anti-androgen (NSAA) medication in addition to castration (medical or surgical) or to castration alone, and (2) reported overall survival, progression-free survival, cancer-specific survival, and/or adverse events. Eligibility was assessed by two independent reviewers. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes were extracted by two independent reviewers using a standardized form. The main outcome measure for comparing effectiveness was overall survival at 1, 2, and 5 years. Secondary outcome measures included progression-free survival and cancer-specific survival. The relationship of specific NSAA on outcome was evaluated. Additionally, the incidence of adverse effects was measured. MAIN RESULTS: Twenty trials enrolling 6,320 patients were included. The pooled OR for overall survival was 1.03 (95% CI:0.85 to 1.25), 1.16 (95% CI:1.00 to 1.33), and 1.29 (95% CI:1.11 to 1.50) at 1, 2, and 5 years respectively. Overall survival was only significant at 5 years. The risk difference at 5 years was 0.048 (95% CI:0.02 to 0.077) and NNT at 5 years 20.8. Progression-free survival was improved only at 1 year follow-up (OR=1.38) and cancer-free survival was improved only at 5 years (OR=1.22). Adverse events occurred more frequently in those assigned to MAB and resulted in withdrawal in 10%. Quality of life was measured in only one study favored orchiectomy alone (less diarrhea and better emotional functioning in the first 6 months). REVIEWER'S CONCLUSIONS: MAB produces a modest overall and cancer-specific survival at 5 years but is associated with increased adverse events and reduced quality of life. PMID- 10796805 TI - Nutritional support for head-injured patients. AB - BACKGROUND: Head injury increases the body's metabolic responses, and therefore nutritional demands. Provision of an adequate supply of nutrients is associated with improved outcome. The best route for administering nutrition (parenterally (TPN) or enterally (EN)), and the best timing of administration (e.g early versus late) of nutrients needs to be established. OBJECTIVES: To quantify the effect on mortality and morbidity of alternative strategies of providing nutritional support following head injury. SEARCH STRATEGY: Trials were identified by computerised searches of the Injuries Group specialised register, Cochrane Controlled Trials Register, MEDLINE, EMBASE and BIDS Index to Scientific and Technical Proceedings, and by checking the reference lists of trials and review articles. SELECTION CRITERIA: Randomised controlled trials of nutritional support following acute traumatic brain injury. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted data and assessed trial quality. Information was collected on death, disability, and incidence of infection. If trial quality was unclear, or if there were missing outcome data, trialists were contacted in an attempt to get further information. MAIN RESULTS: The timing of support: early versus delayed Of the six trials addressing the timing of support, data on mortality were obtained for all six trials, with a total of 257 patients. The relative risk (RR) for death with early nutritional support was 0.71 (95% CI 0.43 to 1.16). Data on disability were available for three trials. The RR for death or disability at the end of follow-up was 0.75 (0.50 to 1.11). The route of feeding: parenteral versus enteral Six trials compared parenteral versus enteral nutrition. Because early support often involves parenteral nutrition, three of the six trials are also included in the previous analyses. Five trials (207 participants) reported mortality. The RR for mortality at the end of follow-up period was 0.66 (0.41 to 1.07). Two trials provided data on death and disability. The RR was 0.69 (95%Cl 0.40 to 1.19). 3. Enteral nutrition: jejunal versus gastric. There was one trial with no deaths and the RR is not estimable. REVIEWER'S CONCLUSIONS: This review suggests that early feeding may be associated with a trend towards better outcomes in terms of survival and disability. Further trials are required. These trials should report not only nutritional outcomes but also the effect on death and disability. PMID- 10796806 TI - Surgery for nonarteritic anterior ischemic optic neuropathy. AB - BACKGROUND: Nonarteritic ischemic optic neuropathy is characterized by sudden and painless loss of vision in one eye, accompanied by pallid swelling of the optic disc. Although various medical interventions, such as corticosteroids and phenytoin sodium, have been used to treat nonarteritic ischemic optic neuropathy, no therapy has been proven effective. OBJECTIVES: The objective of this review is to assess the safety and efficacy of surgical treatment compared with other treatment or usual care in people with nonarteritic ischemic optic neuropathy. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register - Central and MEDLINE. The most recent searches were performed in December 1997. SELECTION CRITERIA: We included randomized trials comparing surgery to no surgery in people with nonarteritic ischemic optic neuropathy. DATA COLLECTION AND ANALYSIS: We obtained full copies of all potentially relevant articles. Only one article described a randomized trial of surgery and it was eligible for inclusion. No formal assessment of quality was done. One reviewer extracted data. No synthesis was required, as there was only one trial. MAIN RESULTS: The one trial identified randomized 258 patients. The only published report with outcomes data for that trial presents preliminary results from 244 patients who had achieved six months of follow-up at the time of the report. Participants assigned to surgery did no better than participants assigned to careful follow-up regarding improved visual acuity of three or more lines of vision at six months: 32.6% of the surgery group improved compared with 42.7% of the careful follow-up group. The adjusted odds ratio (OR), adjusted for baseline visual acuity and diabetes, comparing the two groups for three or more lines improvement was 0.74 (95% confidence interval (CI) 0.39 to 1. 38) (surgery group improvement was worse than careful follow-up). In addition, participants receiving surgery had a significantly greater risk of losing three or more lines of vision at six months: 23.9% in the surgery group worsened compared with 12.4% in the careful follow-up group. The six-month adjusted OR comparing the two groups for loss of three or more lines of vision was 1.96 (95% CI 0.87 to 4.41). Spontaneous improvement of three or more lines of vision was observed in 42.7% of participants in the careful follow-up group. REVIEWER'S CONCLUSIONS: Results from the Ischemic Optic Neuropathy Decompression Trial indicate that optic nerve decompression surgery for nonarteritic ischemic optic neuropathy is not effective. PMID- 10796807 TI - Recordings or summaries of consultations for people with cancer. AB - OBJECTIVES: Many people find it difficult to remember information provided during medical consultations. One way of improving this may be to provide a record of the conversation. This review examined the effects of providing recordings or summaries of their consultations to people with cancer and their families. SEARCH STRATEGY: The following sources were searched: Cochrane Library (to issue 1 of 1999); and MEDLINE; CINAHL; Sociofile; Cancerlit; Dissertation Abstracts; EMBASE; IAC Health & Wellness; JICST; Pascal; ERIC; ASSIA; PsycINFO; Linguistics and Language Behavior Abstracts; Mental Health Abstracts; AMED; CAB Health; DHSS Data; MANTIS (to mid 1998). SELECTION CRITERIA: Randomised and non-randomised controlled trials that evaluate the effects of providing recordings (e.g. audiotapes) or summaries (e.g. letter with reminders of key points) of consultations to people with cancer or their families. Two reviewers assessed studies for inclusion. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and checked by another reviewer. The quality of studies was assessed on six criteria. MAIN RESULTS: Eight studies satisfied the selection criteria. All involved adult participants. The studies did not all measure similar outcomes. In seven studies, between 83% and 96% of participants found recordings or summaries of their consultations valuable. Four out of six studies reported better recall of information for those receiving recordings or summaries. Two out of four studies found that participants provided with a recording or summary were more satisfied with the information received. No studies (out of six) found any statistically significant effect on anxiety or depression. No study evaluated the effects on survival or quality of life. REVIEWER'S CONCLUSIONS: The provision of recordings or summaries of key consultations may benefit most adults with cancer. Although more research is needed to improve our understanding of these interventions, most patients find them very useful. Practitioners should consider offering people tape recordings or written summaries of their consultations. PMID- 10796808 TI - Surgical treatments for ingrowing toenails. AB - OBJECTIVES: To evaluate the effectiveness of methods of the surgical treatment of ingrowing toenails. SEARCH STRATEGY: Electronic database searching (CENTRAL, MEDLINE, EMBASE, CINAHL) followed by investigation of reference lists of the papers identified from the initial search. SELECTION CRITERIA: Any randomised (or quasi-randomised) controlled trial which compares one form of surgical removal of all or part of a toenail due to its impact on the soft tissues to another or others. Studies must have a minimum follow period of six months and aim to permanently remove the troublesome portion of the nail. DATA COLLECTION AND ANALYSIS: Data extraction was carried out independently by the two reviewers using a pre-derived data extraction form and entered into RevMan. Categorical outcomes were analysed as odds ratios with 95% confidence intervals. MAIN RESULTS: Avulsion with phenol versus surgical excision Phenolisation combined with simple avulsion of a nail is more effective than the use of more invasive excisional surgical procedures to prevent symptomatic recurrence at six months or more (OR 0.44 CI 95% 0.24 - 0.80). Avulsion with phenol versus avulsion without phenol The addition of phenol, when performing a total or partial nail avulsion dramatically reduces the rate of symptomatic recurrence, (OR 0.07 95% CI 0.04 - 0.12). This is offset by a significant increase in the rate of post-operative infection when phenol is used (OR 5.69 95% CI 1.93 - 16.77). REVIEWER'S CONCLUSIONS: The evidence suggests that simple nail avulsion combined with the use of phenol, compared to surgical excisional techniques without the use of phenol, is more effective at preventing symptomatic recurrence of ingrowing toenails. The addition of phenol when simple nail avulsion is performed dramatically decreases symptomatic recurrence, but at the cost of increased post operative infection. PMID- 10796809 TI - Palliative chemotherapy for advanced or metastatic colorectal cancer. Colorectal Meta-analysis Collaboration. AB - BACKGROUND: Despite the increasing use of palliative chemotherapy for advanced colorectal cancer, there remains uncertainty as to the true effectiveness of this intervention. This review was therefore undertaken to assess the available evidence for the benefit of palliative chemotherapy in this disease. OBJECTIVES: To determine the benefits and harms of palliative chemotherapy in patients with locally advanced or metastatic colorectal cancer. A secondary objective was to investigate outcomes for younger and elderly patients. SEARCH STRATEGY: Trials were identified by computerised and hand searches of the literature, scanning references and contacting investigators. SELECTION CRITERIA: All randomised controlled trials of palliative chemotherapy compared with supportive care alone in patients with advanced or metastatic colorectal cancer. Both randomised and non-randomised studies were considered when searching for data on quality of life, resource use and cost effectiveness of palliative chemotherapy. DATA COLLECTION AND ANALYSIS: Investigators from all eligible studies were asked to supply individual patient data. Meta-analysis was performed using both published data and individual patient data. Studies were grouped according to whether chemotherapy was administered regionally or systemically. MAIN RESULTS: 13 randomised controlled trials representing a total of 1365 randomised patients met the inclusion criteria. Meta-analysis of a subset of trials that provided individual patient data demonstrated that palliative chemotherapy was associated with a 35% (95% CI 24% to 44%) reduction in the risk of death. This translates into an absolute improvement in survival of 16% at both 6 months and 12 months and an improvement in median survival of 3.7 months. The overall quality of evidence relating to treatment toxicity, symptom control and quality of life was poor. REVIEWER'S CONCLUSIONS: Chemotherapy is effective in prolonging time to disease progression and survival in patients with advanced colorectal cancer. The survival benefit may be underestimated by this meta-analysis, as a proportion of patients in the control arms of some trials received chemotherapy. No age related differences were found in the effectiveness of chemotherapy, but elderly patients were under represented in trials. Treatment toxicity and impact upon quality of life and symptom control have been inadequately assessed in the majority of trials and further research is needed to clarify the palliative benefit of chemotherapy. PMID- 10796810 TI - Single dose paracetamol (acetaminophen), with and without codeine, for postoperative pain. AB - BACKGROUND: Patient surveys have shown that postoperative pain is often not managed well, and there is a need to assess the efficacy and safety of commonly used analgesics as newer treatments become available. Paracetamol (acetaminophen) is an important non-opiate analgesic, commonly prescribed, as well as being available for retail sale. This review seeks to examine the efficacy of paracetamol alone and in combination with codeine, and also considers adverse effects. OBJECTIVES: To assess the analgesic efficacy and adverse effects of a single dose of oral paracetamol (acetaminophen) alone and in combination with codeine for moderate to severe postoperative pain. SEARCH STRATEGY: Published trials were identified from: Medline (1966 to May 1996), Embase (1980 to 1996), Cochrane Library (Issue 2 1996) and the Oxford Pain Relief Database (1950 to 1994). Additional trials were identified from reference lists of retrieved studies. Date of most recent searches: July 1998. SELECTION CRITERIA: Inclusion criteria were: full journal publication, postoperative pain, postoperative oral administration, adult patients, baseline pain of moderate to severe intensity, double-blind design, and random allocation to treatment groups which compared paracetamol with placebo or a combination of paracetamol and codeine with either placebo or the same dose of paracetamol alone. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers, and trials were quality scored. Summed pain intensity and pain relief data were extracted and converted into dichotomous information to yield the number of patients with at least 50% pain relief. This was used to calculate the relative benefit and number-needed-to treat (NNT) for one patient to achieve at least 50% pain relief over 4 to 6 hours compared with placebo. Adverse effects were used to calculate relative risk and number-needed-to-harm (NNH). MAIN RESULTS: We found 40 trials of paracetamol against placebo (4171 patients), 22 trials of paracetamol plus codeine against placebo (1407 patients) and 12 trials of paracetamol plus codeine against the same dose of paracetamol (794 patients). In postoperative pain paracetamol 1000 mg had an NNT of 4.6 (3.8-5.4) for at least 50% pain relief when compared with placebo, and paracetamol 600/650 mg had an NNT of 5.3 (4.1-7.2). Paracetamol 600/650 mg plus codeine 60 mg had an NNT of 3. 6 (2.9-4.5). Comparing paracetamol plus codeine 60 mg with the same dose of paracetamol alone gave an NNT of 7.7 (5.1-17) for at least 50% pain relief. Adverse effects: Relative risk estimates for paracetamol 600/650 mg plus codeine 60 mg versus placebo showed a significant difference for 'drowsiness'/somnolence (NNH 11 (7.5- 0)) and dizziness (NNH 27 (15-164)) but no significant difference for nausea/vomiting. REVIEWER'S CONCLUSIONS: Paracetamol is an effective analgesic with a low incidence of adverse effects. The addition of codeine 60 mg to paracetamol produces additional pain relief even in single oral doses, but may be accompanied by an increase in drowsiness and dizziness. PMID- 10796811 TI - Single dose oral ibuprofen and diclofenac for postoperative pain. AB - BACKGROUND: Ibuprofen and diclofenac are two widely used non-steroidal anti inflammatory (NSAID) analgesics. It is therefore important to know which drug should be recommended for postoperative pain relief. This review seeks to compare the relative efficacy of the two drugs, and also considers the issues of safety and cost. OBJECTIVES: To assess the analgesic efficacy of ibuprofen and diclofenac in single oral doses for moderate to severe postoperative pain. SEARCH STRATEGY: Randomised trials were identified by searching Medline (1966 to December 1996), Embase (1980 to January 1997), the Cochrane Library (Issue 3 1996), Biological Abstracts (January 1985 to December 1996) and the Oxford Pain Relief Database (1950 to 1994). Date of the most recent searches: July 1998. SELECTION CRITERIA: The inclusion criteria used were: full journal publication, postoperative pain, postoperative oral administration, adult patients, baseline pain of moderate to severe intensity, double-blind design, and random allocation to treatment groups which compared either ibuprofen or diclofenac with placebo. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers, and trials were quality scored. Summed pain relief or pain intensity difference over four to six hours was extracted, and converted into dichotomous information yielding the number of patients with at least 50% pain relief. This was then used to calculate the relative benefit and the number-needed-to-treat (NNT) for one patient to achieve at least 50% pain relief. MAIN RESULTS: Thirty-four trials compared ibuprofen and placebo (3,591 patients), six compared diclofenac with placebo (840 patients) and there were two direct comparisons of diclofenac 50 mg and ibuprofen 400 mg (130 patients). In postoperative pain the NNTs for ibuprofen 200 mg were 3.3 (95% confidence interval 2.8 to 4.0) compared with placebo, for ibuprofen 400 mg 2.7 (2.5 to 3.0), for ibuprofen 600 mg 2.4 (1.9 to 3.3), for diclofenac 50 mg 2.3 (2.0 to 2.7) and for diclofenac 100 mg 1.8 (1.5 to 2.1). Direct comparisons of diclofenac 50 mg with ibuprofen 400 mg showed no significant difference between the two. REVIEWER'S CONCLUSIONS: Both drugs work well. Choosing between them is an issue of dose, safety and cost. PMID- 10796812 TI - Primary angioplasty versus intravenous thrombolysis for acute myocardial infarction. AB - BACKGROUND: Intravenous thrombolytic therapy is the standard care for patients with acute myocardial infarction, based upon its widespread availability and ability to reduce patient mortality well demonstrated in randomised trials. Despite its proven efficacy, thrombolytic therapy has limitations. Many patients are ineligible for treatment with thrombolytics. Of those given thrombolytic therapy, 10 to 15 percent have persistent occlusion or reocclusion of the infarct related artery. Consequently, primary angioplasty (primary PTCA) has been advocated as a better treatment of myocardial infarction. OBJECTIVES: To determine whether primary coronary angioplasty is superior to thrombolytic therapy for the treatment of patients with acute myocardial infarction. SEARCH STRATEGY: Electronic search of The Cochrane Library (1998; Issue 2). MEDLINE (to January 1998); references from reviews, trials and previously published meta analyses; and experts. Date of most recent searches January 1998. SELECTION CRITERIA: All unconfounded, randomised controlled trials comparing primary angioplasty against intravenous thrombolysis in patients with acute myocardial infarction DATA COLLECTION AND ANALYSIS: At least two independent reviewers abstracted data on morbidity and mortality and trial characteristics. The following outcomes were assessed: total mortality at the end of the study, reinfarction, stroke of any type, composite endpoint of death and reinfarction, recurrent ischemia, severe bleeding and coronary artery bypass grafting. MAIN RESULTS: Ten trials including 2573 subjects were identified. Compared to thrombolytic therapy, primary angioplasty was associated with a significant reduction in short-term mortality at the end of the studies (relative reduction in risk RRR = 32% 95%CI = 5%;50%). Similar reductions were observed for the rate of reinfarction (RRR = 52%, 95%CI = 30%;67%), recurrent ischemia (RRR = 54%; 95%CI = 39%, 66%) and for the combined criteria death or reinfarction (RRR = 46%; 95%CI=30%;58%). The frequency of strokes of any cause was significantly decreased by 66% (95%CI=28%;84%). No significant difference was observed for the incidence of major bleeding (relative risk RR =1.18, 95%CI = 0.73;1.90) but the confidence interval was large. The superiority of the primary angioplasty over thrombolysis in terms of the composite endpoint (mortality and reinfarction) was less with accelerated t-PA (RR=0.70, 95%CI=0.51;0. 97) than with streptokinase (RR=0.30, 95%CI=0.17;0.53). The biggest and most recent trial, Gusto 2B ( approximately approximately GUSTO-2B 97 approximately approximately ), which involved general as well as higly specialised centres, obtained less favorable results. REVIEWER'S CONCLUSIONS: This meta-analysis suggests that angioplasty provides a short-term clinical advantage over thrombolysis which may not be sustained. Primary angioplasty when available promptly at experienced centres, may be considered the preferred strategy for myocardial reperfusion. In most situations, however, optimal thrombolytic therapy should still be regarded as an excellent reperfusion strategy. PMID- 10796813 TI - Multiple risk factor interventions for primary prevention of coronary heart disease. AB - BACKGROUND: Primary prevention programmes in many countries attempt to reduce mortality and morbidity due to coronary heart disease through risk factor modification. It is widely believed that multiple risk factor intervention using counselling and educational methods is efficacious and cost-effective and should be expanded. Recent trials examining risk factor changes have cast considerable doubt on the effectiveness of these multiple risk factor interventions. OBJECTIVES: To assess the effects of multiple risk factor intervention for reducing cardiovascular risk factors, total mortality, and mortality from coronary heart disease among adults without clinical evidence of established cardiovascular disease. SEARCH STRATEGY: We searched Medline using a standard randomised trial filter. Date of the most recent search was April 1995. SELECTION CRITERIA: Intervention studies using counseling or education to modify more than one cardiovascular risk factor in adults from general populations, occupational groups, or high risk groups. Trials of less than 6 months duration were excluded. DATA COLLECTION AND ANALYSIS: Both reviewers, independently extracted data. Investigators were contacted to obtain missing information. MAIN RESULTS: A total of 18 trials were found of which 10 reported clinical event data. Net changes in systolic and diastolic blood pressure, smoking prevalence, and blood cholesterol were -3.9mmHg (95% CI -4.2,-3.6 mmHg), -2.9 mmHg (95% CI -3.1,-2.7 mmHg), -4.2% (95% CI -4.8, -3.6%), and -0.08mMol/l (95% CI -0.1.-0.06 mMol/l) respectively. In the ten trials with clinical event end-points, the pooled odds ratios for total and coronary heart disease mortality were 0.97 (95% confidence intervals (CI) 0.92-1.02) and 0.97 (95% CI 0.88-1.04) respectively. Statistical heterogeneity between the studies with respect to mortality and risk factor changes was due to trials focusing on hypertensive participants and those using considerable amounts of drug treatment. Only these trials demonstrating significant reductions in mortality. REVIEWER'S CONCLUSIONS: The pooled effects suggest multiple risk factor intervention has no effect on mortality. However, a small, but potentially important, benefit of treatment (about a 10% reduction in CHD mortality) may have been missed. Risk factor changes were relatively modest, were related to the amount of pharmacological treatment used, and in some cases may have been over estimated because of regression to the mean effects, lack of intention to treat analyses, habituation to blood pressure measurement, and use of self-reports of smoking. Interventions using personal or family counseling and education with or without pharmacological treatments appear to be more effective at achieving risk factor reduction and consequent reductions in mortality in high risk hypertensive populations. The evidence suggests that such interventions have limited utility in the general population. PMID- 10796814 TI - Vaccines for preventing influenza in people with cystic fibrosis. AB - BACKGROUND: Viral respiratory tract infections in patients with cystic fibrosis have a deteriorating effect on their lung function and disease progression. Annual influenza vaccination is therefore commonly recommended for patients with cystic fibrosis. OBJECTIVES: To assess the effectiveness of influenza vaccination for patients with cystic fibrosis. SEARCH STRATEGY: We have used the Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, hand searching relevant journals and abstract books of conference proceedings and communication to the drug companies marketing influenza vaccines. SELECTION CRITERIA: All randomised and pseudorandomised trials (published or unpublished) comparing any influenza vaccine with a placebo or with another type of influenza vaccine. DATA COLLECTION AND ANALYSIS: Both reviewers independently assessed trial quality and extracted data. Additional information were obtained by contacting the investigators when it was indicated. MAIN RESULTS: Three trials enrolling a total of 115 patients with cystic fibrosis (76 (66%) were children aged one to 13 years) were included in this review. There was no study comparing a vaccine to a placebo or a whole virus vaccine to a sub unit or split virus vaccine. Two studies compared an intranasal applied live vaccine to an intramuscular inactivated vaccine and one study compared a split virus to a sub unit vaccine (both intramuscular). The incidence of all reported adverse events was high depending on the type of influenza vaccine. The total adverse event rate ranged from 48/201 (24%) for the intranasal live vaccine to 13/30 (43%) for the split virus vaccine. With the limitation of a statistical low power there was no significant difference between the study vaccinations. None of the events were severe. All study influenza vaccinations generated a satisfactory serological antibody response. No trial reported other clinically important benefits. REVIEWER'S CONCLUSIONS: There is currently no evidence from randomised studies that influenza vaccine given to patients with cystic fibrosis is of benefit to them. There is a need for a well constructed clinical study, that assesses the effectiveness of influenza vaccination on important clinical outcome measures. PMID- 10796815 TI - Anterior vaginal repair for urinary incontinence in women. AB - BACKGROUND: Anterior vaginal repair (anterior colporrhaphy) is an operation traditionally used for moderate or severe stress urinary incontinence in women. About a third of adult women experience urinary incontinence. OBJECTIVES: To determine the effects of anterior vaginal repair (anterior colporrhaphy) on stress or mixed urinary incontinence in comparison with other management options. SEARCH STRATEGY: We searched the Cochrane Incontinence Group's trials register, and the reference lists of relevant articles. Date of most recent search: March 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials that included anterior vaginal repair for the treatment of urinary incontinence. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. One trial investigator was contacted for additional information. MAIN RESULTS: Five trials were identified which included 208 women having an anterior vaginal repair and 400 who received comparison interventions. A single small trial provided insufficient evidence to assess anterior repair in comparison with physical therapy. The performance of anterior repair in comparison with needle suspension appeared similar but clinically important differences could not be confidently ruled out. No trials compared anterior repair with sling or laparoscopic interventions, or compared alternative vaginal operations. Anterior repair was less effective than abdominal retropubic suspension based on patient-reported cure rates in four trials both in the short term (failure rate within first year after anterior repair 48/198, 24% vs 30/266, 11%; RR 2.15, 95% CI 1.4 to 3.28) and long-term (80/193, 41% vs 51/261, 20%; RR 2.25, 95% CI 1.66 to 3.04). There was some evidence from one of these trials that this was reflected in fewer repeat operations for incontinence. These findings held irrespective of the co-existence of prolapse (pelvic relaxation). Although later prolapse operation appeared to be equally common after vaginal or abdominal operation there were too few data to judge this reliably. In respect of the type of abdominal retropubic suspension, most data related to comparisons of anterior repair with Burch colposuspension. The few data describing comparison of anterior repair with the Marshall-Marchetti-Krantz procedure were consistent with those for Burch colposuspension. REVIEWER'S CONCLUSIONS: There were not enough data to allow comparison of anterior vaginal repair with physical therapy or needle suspension for primary urinary stress incontinence in women. Abdominal retropubic suspension appeared to be better than anterior vaginal repair judged on subjective cure rates in four trials, even in women who had prolapse in addition to stress incontinence. The need for repeat incontinence surgery also appeared less after the abdominal operation, but this was only reported in one small trial. However, there was not enough information about post-operative complications and morbidity. PMID- 10796816 TI - Surgery for faecal incontinence in adults. AB - BACKGROUND: Faecal incontinence is a debilitating problem with significant medical, social and economic implications. Treatment options include conservative, non-operative interventions (e.g. pelvic floor muscle training, biofeedback, drugs) and surgical procedures. Surgery is used in selected groupsof people when the structural and functional defects in the pelvic floor muscles or the anal sphincter complex can be corrected mechanically. OBJECTIVES: To assess the effects of established surgical techniques for the treatment of faecal incontinence in adults who do not have rectal prolapse. Our aims were firstly to compare surgical management with non-surgical management and secondly, to compare the various surgical techniques. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register, the Cochrane Colorectal Cancer Group trials register, the Cochrane Controlled Trials Register (Issue 2, 1999), Medline (up to March 1999), Embase (1998 up to January 1999), Sigle (1980 up to December 1996), Biosis (1998 up to March 1999), SCI (1998 up to March 1999), ISTP (1982 up to March 1999) and the reference lists of relevant articles. We specifically hand searched the British Journal of Surgery from 1995 to 1998 and the Diseases of the Colon and Rectum from 1995 to 1998. We also perused the proceedings of the Association of Coloproctology, meeting 1999. Date of the most recent literature searches: March 1999. SELECTION CRITERIA: All randomised or quasi-randomised trials of surgery in the management of adult faecal incontinence (other then surgery for rectal prolapse). DATA COLLECTION AND ANALYSIS: Two reviewers independently selected studies from the literature, extracted data and assessed the methodological quality of eligible trials. The three primary outcome measures were: change or deterioration in incontinence, failure to achieve full continence, and the presence of faecal urgency. MAIN RESULTS: Four trials were included with a total sample size of 110 participants. All trials excluded women with anal sphincter defects detected by endoanal ultrasound examination. No trial included a group managed non-surgically. Two trials (56 participants) compared three approaches to pelvic floor repair (anterior levatorplasty, postanal repair and their combination total pelvic floor repair). One trial (30 participants) evaluated adding plication of the anal sphincter to total pelvic floor repair. The fourth trial (24 participants) compared a neosphincter procedure with total pelvic floor repair. No differences in the primary outcomes were detected, but data were few and inconsistently reported. REVIEWER'S CONCLUSIONS: The small number of relevant trials identified together with their small sample sizes and other methodological weaknesses severely limit the usefulness of this review for guiding practice. It was impossible to identify or refute clinically important differences between the alternative surgical procedures. Larger rigorous trials are needed. PMID- 10796817 TI - Surgery for complete rectal prolapse in adults. AB - BACKGROUND: Complete rectal prolapse is a debilitating condition, which affects both the very young and the elderly and can cause faecal incontinence. The range of surgical methods available to correct the underlying anal sphincter or pelvic floor defects in complete rectal prolapse poses the question about the choice of the best operation. OBJECTIVES: To determine the effects of surgery on the treatment of rectal prolapse in adults. The following specific issues have been addressed: I. Whether surgical intervention is better than no treatment; II. Whether an abdominal approach to surgery is better then a perineal approach; III. Whether one method for performing rectopexy is better than another; IV. Whether laparoscopic access is better than open access for surgery; V. Whether resection should be included in the procedure. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register, the Cochrane Colorectal Cancer Group trials register, the Cochrane Controlled Trials Register (Issue 2, 1999), Medline (up to March 1999), Embase (1998 up to January 1999), Sigle (1980 up to December 1996), Biosis (1998 up to March 1999), SCI (1998 up to March 1999), ISTP (1982 up to March 1999) and the reference lists of relevant articles. We hand searched the British Journal of Surgery 1995-8, and the Diseases of the Colon and Rectum 1995 8. We also searched the proceedings of the Association of Coloproctology, meeting 1999. Date of the most recent searches: March 1999. SELECTION CRITERIA: All randomised or quasi-randomised trials of surgery in the management of rectal prolapse. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected studies from the literature, extracted data and assessed the methodological quality of eligible trials. The three primary outcome measures were number of patients with recurrent rectal prolapse, or residual mucosal prolapse or faecal incontinence. MAIN RESULTS: Eight trials were included with a total of 264 participants. No trial included a group receiving no treatment, or anal encirclement, or Delormes procedure, or laparoscopic suture rectopexy, or laparoscopic resection rectopexy. One trial (20 participants) compared both perineal and abdominal resection rectopexy with pelvic floor repair; four trials (175 participants) compared different types of open rectopexy techniques; one trial (21 participants) compared laparoscopic with open mesh rectopexy; and two trials included comparisons between open resection rectopexy and rectopexy alone. In all comparisons data were few. There were no detectable differences in recurrent prolapse between abdominal and perineal approaches, although there was a suggestion that residual faecal incontinence was less common after abdominal surgery. There were no detectable differences between the methods used for fixation during rectopexy. Division, rather than preservation, of the lateral ligaments was associated with less recurrent prolapse but more post-operative constipation, although these findings were found in small numbers. There were too few data with which to compare laparoscopic with open surgery. Bowel resection during rectopexy was associated with lower rates of constipation, but again numbers were small. REVIEWER'S CONCLUSIONS: The small number of relevant trials identified, and their small sample sizes together with other methodological weaknesses severely limit the usefulness of this review for guiding practice. It was impossible to identify or refute clinically important differences between the alternative surgical operations. Larger rigorous trials are needed to improve the evidence with which to define optimum surgical treatment. PMID- 10796818 TI - Psychosocial versus pharmacological treatments for deliberate self harm. AB - OBJECTIVES: To identify and synthesise the findings from all randomised controlled trials that have examined the effectiveness of treatments of patients who have deliberately harmed themselves. SEARCH STRATEGY: Electronic databases screened: MEDLINE (from 1966-February 1999); PsycLit (from 1974-March 1999); Embase (from 1980-January 1999); The Cochrane Controlled Trials Register (CCTR) No.1 1999. Ten journals in the field of psychiatry and psychology were hand searched for the first version of this review. We have updated the hand search of three specialist journals in the field of suicidal research until the end of 1998. Reference lists of papers were checked and trialists contacted. SELECTION CRITERIA: All RCTs of psychosocial and/or psychopharmacological treatment versus standard or less intensive types of aftercare for patients who shortly before entering a study engaged in any type of deliberately initiated self-poisoning or self-injury, both of which are generally subsumed under the term deliberate self harm. DATA COLLECTION AND ANALYSIS: Data were extracted from the original reports independently by two reviewers. Studies were categorized according to type of treatment. The outcome measure used to assess the efficacy of treatment interventions for deliberate self-harm was the rate of repeated suicidal behaviour. We have been unable to examine other outcome measures as originally planned (e.g. compliance with treatment, depression, hopelessness, suicidal ideation/thoughts, change in problems/problem resolution). MAIN RESULTS: A total of 23 trials were identified in which repetition of deliberate self-harm was reported as an outcome variable. The trials were classified into 11 categories. The summary odds ratio indicated a trend towards reduced repetition of deliberate self-harm for problem-solving therapy compared with standard aftercare (0.70; 0.45 to 1.11) and for provision of an emergency contact card in addition to standard care compared with standard aftercare alone (0.45; 0.19 to 1.07). The summary odds ratio for trials of intensive aftercare plus outreach compared with standard aftercare was 0.83 (0.61 to 1.14), and for antidepressant treatment compared with placebo was 0.83 (0. 47 to 1.48). The remainder of the comparisons were in single small trials. Significantly reduced rates of further self-harm were observed for depot flupenthixol vs. placebo in multiple repeaters (0. 09; 0.02 to 0.50), and for dialectical behaviour therapy vs. standard aftercare (0.24; 0.06 to 0.93). REVIEWER'S CONCLUSIONS: There still remains considerable uncertainty about which forms of psychosocial and physical treatments of self harm patients are most effective, inclusion of insufficient numbers of patients in trials being the main limiting factor. There is a need for larger trials of treatments associated with trends towards reduced rates of repetition of deliberate self-harm. The results of small single trials which have been associated with statistically significant reductions in repetition must be interpreted with caution and it is desirable that such trials are also replicated. PMID- 10796819 TI - Ginkgo biloba extract for age-related macular degeneration. AB - BACKGROUND: Ginkgo is used in the treatment of peripheral vascular disease and 'cerebral insufficiency'. It is thought to have several potential mechanisms of action including increased blood flow, platelet activating factor antagonism and prevention of membrane damage caused by free radicals. Vascular factors and oxidative damage are thought to be two potential mechanisms in the pathology of age-related macular degeneration. OBJECTIVES: The objective of this review is to determine the effect of Ginkgo biloba extract on the progression of age-related macular degeneration. SEARCH STRATEGY: The Cochrane Eyes and Vision Group specialised register, the Cochrane Controlled Trials Register - Central, MEDLINE, reference lists of identified trial reports, and the Science Citation Index were searched. The reviewer contacted Investigators of included studies for additional information. The most recent searches were performed in February 2000. SELECTION CRITERIA: All randomised trials where Ginkgo biloba extract had been compared to control in people with age-related macular degeneration were included. DATA COLLECTION AND ANALYSIS: The reviewer extracted data using a standardised form. The data were verified with the trial investigator. MAIN RESULTS: One published trial was identified. Although a beneficial effect was observed, as only 20 people were enrolled in the trial, and assessment of outcome was not masked, its results must be considered equivocal. Adverse effects and quality of life for people with age-related macular degeneration have not been addressed. One unpublished trial is awaiting translation and assessment. REVIEWER'S CONCLUSIONS: The question as to whether people with age-related macular degeneration should take Ginkgo biloba extract to prevent progression of the disease has not been answered by research to date. PMID- 10796820 TI - Immediate post-abortal insertion of intrauterine devices. AB - OBJECTIVES: To assess the safety and efficacy of IUD insertion immediately after spontaneous or induced abortion. SEARCH STRATEGY: We used Medline, Popline, and EMBASE computer searches, supplemented by review articles and contacts with investigators. SELECTION CRITERIA: We sought all randomized controlled trials that had at least one treatment arm that involved IUD insertion immediately after an induced abortion or after curettage for spontaneous abortion. We identified 12 trials which described random assignment but excluded three from this review. Two of these revealed unethical research conduct, and one used alternate assignment to treatments. DATA COLLECTION AND ANALYSIS: We evaluated the methodological quality of each report and abstracted information onto a data collection form. We focused on gross discontinuation rates (single-decrement life table analysis) for accidental pregnancy, perforation, expulsion, and pelvic inflammatory disease. We entered the data into RevMan 3.1 for analysis of Peto odds ratios. MAIN RESULTS: In large multicenter trials, the TCu 220C device proved superior to either the Lippes Loop D or the Copper 7 IUDs for immediate post-abortal insertion. The Peto OR for discontinuation for pregnancy was 0.4 (95% CI 0.2-0.7) compared with the Lippes Loop D and 0.5 (95% CI 0.3-0.8) compared with the Copper 7. Expulsions were also significantly less common with the TCu 220C than with the other two IUDs. In single-center trials, the Nova T IUD had a signicantly higher discontinuation rate for pregnancy than did the Multiload 250 (OR 4.5; 95% CI 1.0 19.8), while the Nova T had a significantly lower discontinuation rate for pregnancy than did the TCu 200 (OR 0.3; 95% CI 0.1-0.9). Only one trial compared immediate vs. delayed insertion. In this trial, the performance of the Copper 7 IUD inserted immediately after abortion was inferior to that after interval insertion (remote from pregnancy), although the differences were not statistically significant. Addition of copper sleeves to a Lippes Loop D improved its performance, but addition of a topical hydrogel to a Spring Coil did not offer benefit. Overall, rates of perforation and pelvic inflammatory disease were low. REVIEWER'S CONCLUSIONS: Insertion of an IUD immediately after abortion is both safe and practical. This was true for both induced and reported "spontaneous" abortions, many of which may have been induced under clandestine circumstances. IUD expulsion rates were higher after second-trimester abortions than after earlier abortions, so delaying insertion may be advisable after later abortions. Although Copper 7 devices inserted unrelated to pregnancy may perform better than those inserted after abortion, many women who express interest in intrauterine contraception do not return for the scheduled insertion. These women may remain unprotected against unintended pregnancy. However, evidence is inadequate to compare the safety and efficacy of IUD insertion immediately after abortion vs. insertion some weeks later. PMID- 10796821 TI - Antibiotics for incomplete abortion. AB - OBJECTIVES: The value of routine antibiotics before surgical evacuation of the uterus in women incomplete abortion is controversial. In some health centres antibiotic prophylaxis is advised in others antibiotics are only prescribed when there is signs of infection. The objective of this review is to evaluate the effectiveness of routine antibiotic prophylaxis to women with incomplete abortion. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE and Popline. Date of last search: January 1999. SELECTION CRITERIA: Randomised trials comparing a policy of routine antibiotic prophylaxis with no routine prophylaxis were eligible for inclusion. DATA COLLECTION AND ANALYSIS: Data extraction was conducted by two reviewers independently. Trial quality was assessed. MAIN RESULTS: One study involving 140 women was included. A second well conducted trial was excluded because of high losses to follow-up. No differences were detected in postabortal infection rates with routine prophylaxis or control. However, compliance with antibiotic treatment was also low. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate a policy of routine antibiotic prophylaxis to women with incomplete abortion. PMID- 10796822 TI - Radiotherapy for the palliation of painful bone metastases. AB - BACKGROUND: Radiotherapy is used commonly to provide pain relief for painful bone metastases, and there is a perception that of the three-quarters of patients who achieve pain relief, half of these stay free from pain. However, the precise contribution from radiotherapy may be unclear because of difficulties in assessing the numbers of people achieving relief, the extent of relief and its duration, and the influence of other contemporaneous interventions, such as analgesics. OBJECTIVES: To assess pain relief from: 1. localised bone metastases achieved by radiotherapy, comparing the efficacy of different fractionation schedules 2. more generalised metastatic disease achieved by radiotherapy or radioisotopes. SEARCH STRATEGY: Studies were identified by searching Medline (1966 to August 1998), Embase (1980 to 1998), the Cochrane Library (1998 Issue 3) and the Oxford Pain Relief Database (1950 to 1994). SELECTION CRITERIA: The inclusion criteria used were: full journal publication, patients with pain due to bone metastases, and random allocation to a radiotherapeutic intervention (either external irradiation or administration of radioisotopes). DATA COLLECTION AND ANALYSIS: The number of patients achieving complete pain relief and at least 50% at one month were compared with an assumed natural history of 1 in 100 patients achieving pain relief without treatment to obtain the number-needed-to-treat (NNT). Summed pain relief or pain intensity difference over four to six hours was extracted, converted into dichotomous information yielding the number of patients with at least 50% pain relief, and used to calculate the relative benefit and the NNT for one patient to achieve at least 50% pain relief. MAIN RESULTS: Twenty trials reported on 43 different radiotherapy fractionation schedules and eight studies of radioisotopes. Radiotherapy produced complete pain relief at one month in 395/1580 (25%) patients, and at least 50% relief in 788/1933 (41%) patients at some time during the trials. There were no differences in the proportions of patients achieving these outcomes between single or multiple fraction schedules. The number-needed-to-treat (NNT) to achieve complete relief at one month (compared with an assumed natural history of 1 in 100 patients whose pain resolved without treatment) was 4.2 (95% CI 3.7-4.7). No pooled estimates of speed of onset of relief, or of its duration, could be obtained. In the largest trial (759 patients) 52% of those who had complete relief had achieved it within four weeks, and the median duration of complete relief was 12 weeks. For more generalised disease, radioisotopes produced similar analgesic results to external irradiation. Adverse effect reporting was poor. There were no obvious differences between the various fractionation schedules in the incidence of nausea and vomiting, diarrhoea or pathological fractures. REVIEWER'S CONCLUSIONS: Radiotherapy is clearly effective at reducing pain from painful bone metastases. There was no evidence of any difference in efficacy between different fractionation schedules, nor indeed of a dose-response with total dose of radiation. For treatment of generalised bone pain both hemibody irradiation and radioisotopes can reduce the number of painful new sites. PMID- 10796823 TI - Preoperative radiotherapy for esophageal carcinoma. Oeosphageal Cancer Collaborative Group. AB - BACKGROUND: The existing randomized evidence has failed to conclusively demonstrate the benefit or otherwise of preoperative radiotherapy in treating patients with potentially resectable esophageal carcinoma. OBJECTIVES: This meta analysis aimed to assess whether there is benefit from adding radiotherapy prior to surgery and whether or not any pre-defined patient subgroups benefit more or less from preoperative radiotherapy SEARCH STRATEGY: Medline and CancerLit searches were supplemented by information from trial registers and by hand searching relevant meeting proceedings and by discussion with relevant trialists, organisations and industry. SELECTION CRITERIA: Trials were eligible for inclusion in this meta-analysis provided they randomized patients with potentially resectable carcinoma of the esophagus (of any histological type) to receive radiotherapy or no radiotherapy prior to surgery. Trials must have used a randomization method which precluded prior knowledge of treatment assignment and completed accrual by December 1993, to ensure sufficient follow-up by the time of the first analysis (September 1995). DATA COLLECTION AND ANALYSIS: A quantitative meta-analysis using updated data from individual patients from all properly randomized trials (published or unpublished) comprising 1147 patients (971 deaths) from five randomized trials. This approach was used to assess whether preoperative radiotherapy improves overall survival and whether it is differentially effective in patients defined by age, sex and tumour location. MAIN RESULTS: With a median follow-up of 9 years, in a group patients with mostly squamous carcinomas, the hazard ratio (HR) of 0.89 (95% CI 0.78-1.01) suggests an overall reduction in the risk of death of 11% and an absolute survival benefit of 3% at 2 years and 4% at 5 years. This result is not conventionally statistically significant (p=0.062). No clear differences in the size of the effect by sex, age or tumor location were apparent. REVIEWER'S CONCLUSIONS: Based on existing trials, there was no clear evidence that preoperative radiotherapy improves the survival of patients with potentially resectable esophageal cancer. These results indicate that if such preoperative radiotherapy regimens do improve survival, then the effect is likely to be modest with an absolute improvement in survival of around 3 to 4%. Trials or a meta-analysis of around 2000 patients (90% power, 5% significance level) would be needed to reliably detect such an improvement (from 15 to 20%). PMID- 10796824 TI - Tonsillectomy versus non-surgical treatment for chronic / recurrent acute tonsillitis. AB - BACKGROUND: Surgical removal of the tonsils (tonsillectomy) is a common but controversial ENT operation. OBJECTIVES: To determine the effect of tonsillectomy in patients with chronic/recurrent acute tonsillitis. SEARCH STRATEGY: Cochrane Controlled Trials Register, Medline, Embase, bibliographies. SELECTION CRITERIA: Randomised controlled trials comparing tonsillectomy with non-surgical treatment in adults and children with chronic/recurrent acute tonsillitis. Trials which included reduction in the number and severity of tonsillitis and sore throat as main outcome measures. DATA COLLECTION AND ANALYSIS: Two authors applied the inclusion/exclusion criteria independently. MAIN RESULTS: No trials evaluating the effectiveness of tonsillectomy in adults were identified. Two trials from Pittsburgh assessed tonsillectomy in children. Significant baseline differences between the surgical and non-surgical groups and the inclusion of children who also underwent adenoidectomy prevent firm conclusions being drawn from the fully published trial. Limited and insufficient information is available from the second study; further details are awaited. REVIEWER'S CONCLUSIONS: The effectiveness of tonsillectomy has not been formally evaluated. Further trials addressing relevant outcome measure are required. PMID- 10796825 TI - Conservative management of post prostatectomy incontinence. AB - BACKGROUND: Urinary incontinence after prostatectomy is a common problem. Conservative management of this condition includes pelvic floor muscle training, biofeedback, electrical stimulation using a rectal electrode, transcutaneous electrical nerve stimulation, or a combination of methods. OBJECTIVES: To assess the effects of conservative management for urinary incontinence after transurethral, suprapubic, radical retropubic or perineal prostatectomy. SEARCH STRATEGY: The Cochrane Incontinence Group's trials register, Medline, Cinahl, Embase, PsycLit and ERIC all up to January 1999, and reference lists of relevant articles. We contacted investigators to locate studies and we handsearched the following conference proceedings: American Urological Association (1989-1999); Society of Urologic Nurses and Associates (1991-1998); Wound Ostomy and Continence Nurses (1996-1999); and International Continence Society (1980-1998). Date of most recent searches: January 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials which evaluated conservative management aimed at improving urinary continence after prostatectomy. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the methodological quality of studies and abstracted data from included trials onto a standard form. MAIN RESULTS: Only five randomised trials were identified which included 365 men, each evaluating different treatments, and all studying men after radical prostatectomy. The trials were of moderate quality and data were not available for many of the pre stated outcomes. Confidence intervals for both dichotomous and continuous data were wide; it was not possible to reliably identify or rule out a useful effect. Men's symptoms tended to improve over time, irrespective of management. REVIEWER'S CONCLUSIONS: The value of the various approaches to conservative management of post prostatectomy incontinence remains uncertain. Further well designed trials are needed. PMID- 10796826 TI - SSRIs versus other antidepressants for depressive disorder. AB - OBJECTIVES: To examine the relative efficacy of selective serotonin reuptake inhibitors (SSRIs) compared to other antidepressants. SEARCH STRATEGY: The search strategy included a search of (a) Electronic bibliographic databases (MEDLINE, EMBASE); (b) reference lists of related reviews (c) reference lists of all located studies (d) contact with the manufacturer and (e) the Cochrane Group register of controlled trials SELECTION CRITERIA: Randomised controlled trials comparing selective serotonin reuptake inhibitors with other kinds of antidepressants in the treatment of patients with depressive disorders. The outcome measures assessed included measures of the severity of depression. DATA COLLECTION AND ANALYSIS: Data were collected from each study the main outcome measurefrom each study. These included: mean Hamilton depression rating scale, mean Montgomery & Asberg depression rating scale, Clinical Global Impression rating scale. An analysis of standardised mean difference of these scales was performed using Review Manager 3. 1 software. The presence of heterogeneity of treatment effect was assessed MAIN RESULTS: Ninety-eight trials contributed data to the analysis of the relative efficacy of SSRIs and related drugs with comparator antidepressants (Figure 3 & Appendix 3). Analysis of efficacy was based upon 5044 patients treated with an SSRI or related drug, and 4510 treated with an alternative antidepressant. The standardised effect size for SSRIs and related drugs together versus alternative antidepressants using a fixed effects model was 0. 035 (95% CI -0.006 to 0.076; Q = 149.25, df = 97, p < 0.001). REVIEWER'S CONCLUSIONS: There are no clinically significant differences in effectiveness between selective serotonin reuptake inhibitors and tricyclic antidepressants. Treatment decisions need to be based on considerations of relative patient acceptability, toxicity and cost. PMID- 10796827 TI - Helmets for preventing head and facial injuries in bicyclists. AB - BACKGROUND: Each year, in the United states, approximately 900 persons die from injuries due to bicycle crashes and over 500,000 persons are treated in emergency departments. Head injury is by far the greatest risk posed to bicyclists, comprising one-third of emergency department visits, two-thirds of hospital admissions, and three-fourths of deaths. Facial injuries to cyclists occur at a rate nearly identical to that of head injuries. Although it makes inherent sense that helmets would be protective against head injury, establishing the real-world effectiveness of helmets is important. A number of case-control studies have been conducted demonstrating the effectiveness of bicycle helmets. Because of the magnitude of the problem and the potential effectiveness of bicycle helmets, the objective of this review is to determine whether bicycle helmets reduce head, brain and facial injury for bicyclists of all ages involved in a bicycle crash or fall. OBJECTIVES: To determine whether bicycle helmets reduce head, brain and facial injury for bicyclists of all ages involved in a bicycle crash or fall. SEARCH STRATEGY: We searched The Cochrane Controlled Trials Register, MEDLINE, EMBASE, Sport, ERIC, NTIS, Expanded Academic Index, CINAHL, PsycINFO, Occupational Safety and Health, and Dissertations Abstracts. We checked reference lists of past reviews and review articles, studies from government agencies in the United States, Europe and Australia, and contacted colleagues from the International Society for Child and Adolescent Injury Prevention, World Injury Network, CDC funded Injury Control and Research Centers, and staff in injury research agencies around the world. SELECTION CRITERIA: Controlled studies that evaluated the effect of helmet use in a population of bicyclists who had experienced a crash. We required that studies have complete outcome ascertainment, accurate exposure measurement, appropriate selection of the comparison group and elimination or control of factors such as selection bias, observation bias and confounding. DATA COLLECTION AND ANALYSIS: Five published studies met the selection criteria. Two abstractors using a standard abstraction form independently abstracted data. Odds ratios with 95% CI were calculated for the protective effect of helmet for head and facial injuries. Study results are presented individually. Head and brain injury results were also summarized using meta-analysis techniques. MAIN RESULTS: No randomized controlled trials were found. This review identified five well conducted case control studies which met our selection criteria. Helmets provide a 63%-88% reduction in the risk of head, brain and severe brain injury for all ages of bicyclists. Helmets provide equal levels of protection for crashes involving motor vehicles (69%) and crashes from all other causes (68%). Injuries to the upper and mid facial areas are reduced 65%. REVIEWER'S CONCLUSIONS: Helmets reduce bicycle-related head and facial injuries for bicyclists of all ages involved in all types of crashes including those involving motor vehicles. PMID- 10796828 TI - Medical anti-shock trousers (pneumatic anti-shock garments) for circulatory support in patients with trauma. AB - BACKGROUND: Medical antishock trousers (MAST) have been used to increase venous return to the heart until definitive care could be given. This, combined with compression of blood vessels, is believed to cause the movement of blood from the lower body to the brain, heart and lungs. However, the equipment is expensive, and may have adverse effects. OBJECTIVES: To quantify the effect on mortality and morbidity of the use of medical anti-shock trousers (MAST)/ pneumatic anti-shock garments (PASG) in patients following trauma. SEARCH STRATEGY: Trials were identified by searches of the Cochrane Injuries Group Trials Register, the Cochrane Controlled Trials Register, MEDLINE, EMBASE, BIDS ISI Service and Science Citation Index. References in relevant papers identified were followed up. A citation analysis of references to randomised controlled trials was conducted using the Science Citation Index. Authors of identified trials were contacted and asked about any other trials that may have been conducted, whether published or unpublished. SELECTION CRITERIA: Randomised and quasi-randomised trials of MAST/PASG in patients following trauma (excluding fractures of the extremities in which MAST/PASG may be used as a splint). DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Data were collected on mortality, duration of hospitalisation and ICU stay, and quality of allocation concealment. MAIN RESULTS: Two trials were identified that met the inclusion criteria. These trials included 1202 randomised patients in total; however, data for only 1075 of these were available. The relative risk of death with MAST was 1.13 (95% CI 0.97 to 1.32). Duration of hospitalisation and of intensive care unit stay was longer in the MAST treated group. The weighted mean difference in the length of intensive care unit stay was 1.7 days (95% CI 0.33 to 2.98). REVIEWER'S CONCLUSIONS: There is no evidence to suggest that MAST/PASG application reduces mortality, length of hospitalisation or length of ICU stay in trauma patients and it is possible that it may increase these. These data do not support the continued use of MAST/PASG in the situation described. However, it should be recognised that, due to the poor quality of the trials, conclusions should be drawn with caution. PMID- 10796829 TI - Interventions for preventing injuries in problem drinkers. AB - OBJECTIVES: To assess the effect of interventions for problem drinking on subsequent injury risk. SEARCH STRATEGY: Data Sources.- Twelve computerized databases: MEDLINE (1966-8/96), EMBASE (1982-1/97), Cochrane Controlled Trials Register (1997, issue #1), PSYCHINFO (1967-1/97), CINAHL (1982-10/96), ERIC (1966 12/96), Dissertation Abstracts International (1861-11/96), IBSS (1961-1/97), ISTP (1982-1/97) and three specialized transportation databases, using terms for problem drinking combined with terms for controlled trials; bibliographies of relevant trials; and contact with authors and government agencies. SELECTION CRITERIA: Data Selection.- Randomized controlled trials of interventions among particiapnts with problem drinking, which are intended to reduce alcohol consumption or to prevent injuries or their antecedents, and which measured injury-related outcomes. Of 7014 studies identified, 19 (0. 3%) met the inclusion criteria. DATA COLLECTION AND ANALYSIS: Data Extraction.- Two authors extracted data on participants, interventions, follow-up, allocation concealment, and outcomes, and independently rated allocation concealment quality. MAIN RESULTS: Data Synthesis.- In completed trials, interventions for problem drinking were associated with reduced suicide attempts, domestic violence, falls, drinking related injuries, and injury hospitalizations and deaths, with reductions ranging from 27-65%. Several interventions among convicted drunk drivers reduced motor vehicle crashes and injuries. Because few trials were sufficiently large to assess effects on injuries, individual effect estimates were imprecise. We did not combine the results quantitatively because the interventions, patient populations, and outcomes were so diverse. REVIEWER'S CONCLUSIONS: Conclusion.- Interventions for problem drinking may reduce injuries and their antecedents. Because injuries account for much of the morbidity and mortality from problem drinking, further studies are warranted to evaluate the effect of treating problem drinking on injuries. PMID- 10796830 TI - Education for contraceptive use by women after childbirth. AB - OBJECTIVES: Postpartum education on contraceptive use is a routine component of discharge planning in many different countries with a wide variety of health care systems. This education is based on assumptions concerning women's receptivity to contraceptive education during the postpartum period and their presumed lack of access to such education after that time. The objective of this review is to assess the effects of education about contraceptive use to postpartum mothers. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, EMBASE, CINAHL, Psychlit, Popline, citations indexes and reference lists of relevant articles. We contacted subject experts to locate additional research, in addition to the Group's Specialised Register of Controlled Trials. Date of the most recent search: April 1999. SELECTION CRITERIA: Trials using random or quasi random methods of allocation which evaluated the effectiveness of postpartum education about contraceptive use. DATA COLLECTION AND ANALYSIS: Two independent reviewers abstracted data on trial characteristics and results. MAIN RESULTS: Three trials were identified with 5438 women. These trials were conducted in Lebanon, Peru and Nepal. None of the trials examined all major prespecified endpoints. Postpartum education about contraceptive use influenced short-term use assessed between 40 days and three months post-partum. Women in the intervention groups were less likely to be non-users than women in the comparison groups (Odds Ratio (OR) = 0.47, 95% Confidence Interval (CI) 0.39 to 0.58). This benefit was not apparent following analysis of data from better quality studies (OR = 0.67, 95% CI 0.41 to 1.13). An apparent benefit on contraceptive use at six months post partum (OR = 0.52, 95% CI 0.37 to 0.74) was not apparent following sensitivity analyses (OR = 0.59, 95% CI 0.33 to 1.06). Data are inadequate to assess the impact on cessation of breast feeding and non-attendance at family planning clinics. Unplanned pregnancies, knowledge about contraception and satisfaction with care were not assessed in any trial. REVIEWER'S CONCLUSIONS: The effectiveness of postpartum education about contraceptive use has not yet been established in randomised controlled trials. Such education may be effective in increasing the short-term use of contraception. However, there are only limited data examining a more-important longer-term effect on the prevention of unplanned pregnancies. Research needs to be undertaken to assess the effectiveness of the minimalist education provided in more developed countries and the variety of programs provided in less developed regions. Such research should examine the content, timing, range and organisation of postpartum education on contraceptive use, as well as its impact on breast feeding rates. PMID- 10796831 TI - Perioperative local anaesthesia for reducing pain following tonsillectomy. AB - BACKGROUND: Pain relief after tonsillectomy is an important part of post operative management. Sometimes local anaesthetics are administed to the tonsillar region, but their effectiveness in relieving pain has not been formally assessed. OBJECTIVES: To assess the effects of pre- and post-operative local anaesthesia for pain reduction following tonsillectomy. SEARCH STRATEGY: Cochrane Controlled Trials Register, Medline, Embase and the Oxford Pain Database. Reference lists from identified publications, including those in non-English language publications, were scanned. Date of the most recent search was September 1998. SELECTION CRITERIA: Randomised controlled trials of adults and/or children undergoing tonsillectomy alone with local anaesthetic (a) injected into the tonsillar region immediately prior to removal of the tonsils (b) injected into the tonsillar region after removal of the tonsils (c) sprayed or otherwise applied to the tonsillar region after removal of the tonsils Outcome measures included the reduction in demand for post-operative analgesia, both in terms of time elapsed to request for first analgesia and of the total amount of analgesia used. DATA COLLECTION AND ANALYSIS: All three reviewers extracted data independently and assessed trials for quality. Four authors were contacted to obtain information on either the method of randomisation or the type of intervention used. MAIN RESULTS: Thirty trials were initially identified but only six trials met the inclusion criteria. These were all published in the English language literature between 1989 and 1997. Trials were excluded because procedures in addition to tonsillectomy were undertaken, inadequate randomisation techniques were used or because group sizes were inadequate. Of the six included studies, five involved local anaesthetic injection and one application of local anaesthetic spray. These studies used validated pain scores and measurement of supplemental analgesic intake for assessment of pain. However, some studies also included inappropriate, possibly post-hoc, outcome measures such as 'global pain scores' which may have resulted in bias. No included study showed a significant difference between intervention and control, other than for outcome measures which were felt to be inappropriate, such as the one mentioned above. REVIEWER'S CONCLUSIONS: There is no evidence that the use of perioperative local anaesthetic in patients undergoing tonsillectomy improves post-operative pain control. The trials identified were of small size and several involved the perioperative co administration of intravenous opiates which may have masked any beneficial effect of the local anaesthetic. Further randomised controlled trials are necessary. PMID- 10796832 TI - Low protein diets delay end-stage renal disease in non diabetic adults with chronic renal failure. AB - BACKGROUND: Since more than fifty years, low protein diets are proposed to patients with kidney failure. However, the effects of these diets in preventing severe renal failure and the need for maintenance dialysis is still controversial. OBJECTIVES: To determine the efficacy of low protein diets in delaying the need to start maintenance dialysis. SEARCH STRATEGY: Medline and Embase search from January 1966 through June 1999. Congress abstracts (American Society of Nephrology since 1990, European Dialysis Transplant Association since1985, International Society of Nephrology since 1987). Direct contacts with investigators. SELECTION CRITERIA: Randomised trials comparing two different levels of protein intake in adult patients suffering from moderate to severe renal failure, followed for at least one year. Exclusion of patients with diabetic nephropathy. DATA COLLECTION AND ANALYSIS: Seven trials selected over 40 studies since 1975. A total of 1494 patients analysed, 753 receiving a reduced protein intake and 741 a larger protein intake. Collection of the number of "renal death" being the need for starting dialysis, the death of a patient or the transplantation of a kidney during the trials. MAIN RESULTS: 242 renal deaths were recorded, 101 in the low protein diet and 141 in the larger protein diet group, giving an odds ratio of low protein to control of 0.62 with a 95% confidence interval of 0.46 to 0.83 (p<0.001, Peto odds ratio). To spare one extra renal death, 17 patients need to be treated (NNT) with a low protein diet for approximately two years. REVIEWER'S CONCLUSIONS: Reducing protein intake in patients with chronic renal failure do reduce the occurence of renal death by about 40% as compared with larger or unrestricted protein intake and therefore should be warmly recommended to patients. The optimal level of protein intake cannot be deduced from the present work. However, based on other nutritional studies perfomed in patients having chronic renal failure, we recommend a protein intake of 0.6 g/kg/day and an energy intake no less than 35 kcal/kg/day. Skilled dietitian survey should be offered to closely monitor these values and the patient's nutritional status. PMID- 10796833 TI - Progestogens versus oestrogens and progestogens for irregular uterine bleeding associated with anovulation. AB - BACKGROUND: Dysfunctional uterine bleeding (DUB) is "excessively heavy, prolonged or frequent bleeding of uterine origin which is not due to pregnancy or to recognizable pelvic or systemic disease". Anovulation may be inferred from a number of observations but in the normal clinical situation, anovulation is often assumed when a woman presents with heavy, prolonged or frequent bleeding, particularly in those at the extremes of reproductive life and in women known to have the polycystic ovarian syndrome. Menstrual bleeding that is irregular or excessive is usually poorly tolerated by the majority of women. Changes in the length of the menstrual cycle generally imply disturbances of the hypothalamo pituitary-ovarian (HPO) axis. In anovulatory DUB with acyclic (irregular) oestrogen production there will be no progesterone withdrawal from oestrogen primed endometrium and so cycles are irregular. Prolonged oestrogen stimulation may cause a build up of endometrium with erratic bleeding as it breaks down and is expelled. This is the rationale for using cyclical progestogens during the second half of the menstrual cycle in order to provoke a regular withdrawal bleed. Continuous progestogen is intended to induce endometrial atrophy and hence to prevent oestrogen-stimulated endometrial proliferation. Progestogens, and oestrogens and progestogens in combination are already widely used in the management of irregular or excessive bleeding due to DUB, but the regime, dose and type of progestogen used varies widely with little consensus about the optimum treatment approach. OBJECTIVES: To determine the effectiveness and acceptability of progestogens alone, and oestrogens and progestogens in combination in the management of irregular bleeding associated with anovulation. SEARCH STRATEGY: The search strategy of the Menstrual Disorders Group was used to identify all randomised trials of progestogens alone or in combination with oestrogens in the management of irregular menstrual bleeding associated with anovulation. In addition a search of the Cochrane Controlled Trials Register was undertaken. SELECTION CRITERIA: All randomised controlled trials of progestogens (via any route) alone or in combination with oestrogens in the treatment of irregular bleeding associated with anovulation. DATA COLLECTION AND ANALYSIS: Study quality assessment and data extraction were carried out independently by two reviewers. Both reviewers were experts in the content matter. MAIN RESULTS: No randomised trials were identified which compared progestogens with oestrogens and progestogens in the management of irregular bleeding associated with anovulation. Only one small, non-randomised study compared two progestogen regimes in the management of heavy and irregular bleeding in subjects with confirmed anovulation. One randomised study compared the effects of two progestogens on endometrial histology in subjects with a variety of menstrual symptoms, half of whom had cystic glandular hyperplasia. No studies were found which compared progestogens with oestrogens and progestogens in combination or with placebo in the management of irregular bleeding associated with anovulation. REVIEWER'S CONCLUSIONS: There is a paucity of randomised studies relating to the use of progestogens and of oestrogens and progestogens in combination in the treatment of irregular bleeding associated with anovulation. Further research is needed to establish the role of these treatments in the management of this common gynaecological problem. PMID- 10796834 TI - Surgical interruption of pelvic nerve pathways for primary and secondary dysmenorrhoea. AB - BACKGROUND: Dysmenorrhoea is the occurrence of painful menstrual cramps of uterine origin and is a very common gynaecological complaint. Medical therapy for dysmenorrhoea includes oral contraceptive pills (OCP) and nonsteroidal anti inflammatory drugs (NSAIDS) which both act by suppressing prostaglandin levels. While these treatments are very successful there is still a 20-25% failure rate and surgery has been an option for cases of dysmenorrhoea that fail to respond to medical therapy. Uterine nerve ablation (UNA) and presacral neurectomy (PSN) are two surgical treatments that have become increasingly utilised in recent years. These procedures both interrupt the majority of the cervical sensory nerve fibres, thus diminishing uterine pain. Uncontrolled studies have supported the use of these procedures for primary dysmenorrhoea however both operations only partially interrupt some of the cervical sensory nerve fibres in the pelvic area; therefore dysmenorrhoea associated with additional pelvic pathology may not always benefit from this type of surgery. OBJECTIVES: To assess the effectiveness of surgical interruption of pelvic nerve pathways as treatment for primary and secondary dysmenorrhoea, and to determine the most effective surgical treatment. SEARCH STRATEGY: Electronic searches of the Cochrane Menstrual Disorders and Subfertility Group Register of controlled trials, MEDLINE, and EMBASE were performed to identify relevant randomised controlled trials (RCTs). Attempts were also made to identify trials from citation lists of review articles and handsearching. In most cases, the first or corresponding author of each included trial was contacted for additional information. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of surgical techniques of interruption of the pelvic nerve pathways (both open and laparoscopic procedures) for the treatment of primary and secondary dysmenorrhoea. The main outcome measures were pain relief and adverse effects. DATA COLLECTION AND ANALYSIS: Seven RCTs were identified that fulfilled the inclusion criteria for this review. One trial (Sutton 1994) was excluded because another treatment was given in combination with destruction of pelvic nerve pathways and the effects of these two treatments could not be separated. Of the remaining six trials, three were included in the meta-analysis (Chen 1996, Candiani 1992, Lichten 1987). The results of the other three trials (Dover 1999, Tjaden 1990, Vercellini 1997) were included in the text of the review for discussion because the data were not available in a form that allowed them to be combined in a meta-analysis. MAIN RESULTS: For the treatment of primary dysmenorrhoea there is some evidence of the effectiveness of uterine nerve ablation (UNA) when compared to a control of no treatment. The comparison between UNA with presacral neurectomy (PSN) for primary dysmenorrhoea showed no significant difference in pain relief in the short term, however long term PSN was shown to be significantly more effective. For the treatment of secondary dysmenorrhoea the identified RCTs addressed only endometriosis. The treatment of UNA combined with surgical treatment of endometrial implants versus surgical treatment of endometriosis alone showed that the addition of UNA did not aid pain relief. For PSN combined with endometriosis treatment versus endometriosis treatment alone there was also no overall difference in pain relief, although the data suggests a significant difference in relief of midline abdominal pain. Adverse events were significantly more common for presacral neurectomy, however the majority were complications such as constipation, which may spontaneously improve. REVIEWER'S CONCLUSIONS: There is insufficient evidence to recommend the use of nerve interruption in the management of dysmenorrhoea, regardless of cause. Future RCTs should be undertaken. PMID- 10796835 TI - Postoperative procedures for improving fertility following pelvic reproductive surgery. AB - BACKGROUND: Hydrotubation with oil-soluble contrast media for unexplained infertility and adhesiolysis for infertility due to peritubal adhesions are primary procedures of recognised benefit. It is less clear whether postoperative procedures such as hydrotubation or second-look laparoscopy with adhesiolysis are beneficial following pelvic reproductive surgery. OBJECTIVES: To assess the value of postoperative hydrotubation and second-look laparoscopy with adhesiolysis following female pelvic reproductive surgery. SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility Group was used for the identification of all relevant randomised controlled trials. SELECTION CRITERIA: All trials where a postoperative procedure following pelvic reproductive surgery was compared with a control group generated by randomisation were considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Five randomised controlled trials were identified and included in this review, after an attempt to obtain further information from authors of all five trials. All trials were assessed for quality criteria. The studied outcomes were pregnancy, live birth, ectopic pregnancy and miscarriage rates and the rates of tubal patency and procedure related complications. Reviewers extracted the data independently and odds ratios for these dichotomous outcomes were estimated from the data. MAIN RESULTS: The odds of pregnancy, live birth, ectopic pregnancy and miscarriage were not significantly different with postoperative hydrotubation versus no hydrotubation nor with second-look laparoscopy and adhesiolysis versus no second-look laparoscopy. Whether hydrotubation was early or late and whether hydrotubation fluid contained steroid or not had no significant impact on the odds of pregnancy, live birth, ectopic pregnancy or miscarriage. The odds of pregnancy and live birth were significantly increased and infective complications significantly decreased by hydrotubation with fluid containing antibiotic versus hydrotubation with fluid containing no antibiotic, in late hydrotubation following tubal stent removal six weeks after tubal surgery. The odds of at least one patent fallopian tube were significantly increased with late hydrotubation following tubal stent removal versus early hydrotubation in women who had no tubal stenting, but this intervention had no significant impact on the odds of pregnancy, live birth, ectopic pregnancy or miscarriage. REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the routine practice of hydrotubation or second-look laparoscopy following female pelvic reproductive surgery. The studies on which this conclusion is based were either poor quality or underpowered. These interventions should be performed in the context of a good quality, adequately powered randomised controlled trial. Postoperative hydrotubation with fluid containing antibiotic following tubal surgery may offer benefit over hydrotubation fluid without antibiotic. A randomised controlled trial of postoperative hydrotubation with antibiotic-containing fluid versus no hydrotubation for improving fertility following tubal surgery is justified. PMID- 10796836 TI - Prophylactic antibiotics for cystic fibrosis. AB - BACKGROUND: Patients with cystic fibrosis are sometimes prescribed antibiotics to take continuously on a prophylactic (preventative) basis. This approach is most commonly used in infants where the objective is to reduce pulmonary infection with Staphylococcus aureus and prevent lung damage. This approach may also be used in older patients. This review evaluates the evidence for the effectiveness of this approach and considers potential adverse effects. OBJECTIVES: To compare continuous oral antibiotic prophylaxis with no prophylaxis (short courses of oral antibiotics given as clinically indicated) in patients with cystic fibrosis. This review considers both the effectiveness of prophylaxis (bacteria isolated from the respiratory tract, requirement for additional antibiotic treatment, lung function, survival) and the adverse effects. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group clinical trials register was used. This comprises references identified from a comprehensive search of electronic databases, as well as hand searching relevant journals and conference abstracts. Companies manufacturing anti-staphylococcal antibiotics were also approached for unpublished data. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudo-randomised trials where continuous oral prophylactic antibiotics, given for a period of at least one year, were compared to intermittent antibiotic therapy given "as required." Cystic fibrosis patients of any disease severity were considered. DATA COLLECTION AND ANALYSIS: Trials were assessed for eligibility, methodological quality and data extraction by two reviewers (AS & SW). The following outcomes were assessed: lung function; nutrition (weight standard deviation score); survival; requirement for additional antibiotic treatment; isolates of pathogens from the respiratory tract; occurrence of adverse reactions to prophylactic antibiotics. MAIN RESULTS: Only two studies, totalling 66 patients (over half of whom were infants), were suitable for inclusion in the review. A reduced prevalence of Staphylococcus aureus in the respiratory secretions was seen in children receiving anti-staphylococcal antibiotic prophylaxis, although no effect was seen on other common pathogens. This is associated with a reduced requirement for additional courses of oral antibiotics and fewer hospital admissions in the first two years of life in patients receiving prophylaxis. No effect on infant lung function has been shown after one year of prophylactic treatment. Data are not available on adverse effects of the interventions. As the duration of the studies reviewed has been of two years or less, conclusions cannot be drawn about the long term effects of prophylaxis on acquisition of Pseudomonas aeruginosa and survival. REVIEWER'S CONCLUSIONS: Anti-staphylococcal antibiotic prophylaxis may be of benefit when commenced early in infancy and continued up to two years of age. There is insufficient evidence from this review to say whether use in older children, or adults, or for periods of over two year is beneficial. PMID- 10796837 TI - Neonatal screening for sickle cell disease. AB - BACKGROUND: Sickle cell disease is an inherited disorder that occurs throughout the world with its highest incidence in areas of Africa where malaria is endemic. It affects up to one in 60 infants born in some areas of Africa. There are a number of potentially serious complications associated with the condition, and it is suggested that early treatment (before symptoms develop) can improve both morbidity and mortality. Screening for the condition in the neonatal period would enable early diagnosis and therefore early treatment. OBJECTIVES: To assess whether there is evidence that neonatal screening for sickle cell disease rather than symptomatic diagnosis reduces adverse short and long term outcomes for those in whom the disease is detected, without adverse outcomes in the population screened. SEARCH STRATEGY: We searched the Controlled Trials Register of the Cochrane Cystic Fibrosis and Genetic Disorders Group (See Group search strategy). Contact was made with experts in the field for any work as yet unpublished and reference lists of published studies were also searched. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: Any randomised or pseudorandomised trial, published or unpublished comparing diagnosis by screening to clinical diagnosis would have been considered eligible for inclusion. DATA COLLECTION AND ANALYSIS: No trials of neonatal screening for sickle cell disease were found. MAIN RESULTS: No trials of neonatal screening for sickle cell disease were found. REVIEWER'S CONCLUSIONS: There is a lack of evidence from trials of neonatal screening for sickle cell disease. There is evidence of benefit from early treatment which is made possible by screening and there are a number of reviews and economic analyses of non-trial literature suggesting that screening is appropriate. Health care providers must therefore assess whether the information provided by these documents is relevant to their practice and situation when making decisions regarding neonatal screening for sickle cell disease. Systematic reviews of early treatments/interventions, including penicillin prophylaxis, pneumococcal vaccine and parental education should be considered. PMID- 10796838 TI - Inhaled corticosteroids for cystic fibrosis. AB - BACKGROUND: Maintenance of optimal lung function is an important therapeutic goal in cystic fibrosis as it is lung damage that, in the long term, is responsible for most premature death among affected people. Inhaled corticosteroids are being increasingly used to treat children and adults with cystic fibrosis. The rationale for their use is that they have the potential to reduce lung damage arising from inflammation. However chronic use of inhaled steroids may also have adverse effects. It is thus important to establish the current level of evidence about the potential benefits and harms of this practice. OBJECTIVES: The objective of this review is to assess the effectiveness of regular use of inhaled corticosteroids when compared to no inhaled corticosteroids, in the management of patients with cystic fibrosis. SEARCH STRATEGY: Trials were ascertained from the Cochrane Cystic Fibrosis and Genetic Disorders Specialised Register of Controlled Trials which includes published and unpublished trials identified through electronic databases such as Medline and Embase as well as those identified from handsearching of journals and conference proceedings. Pharmaceutical companies manufacturing inhaled corticosteroids were also contacted to identify any trials of inhaled corticosteroids in cystic fibrosis. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All trials, both published and unpublished, in which inhaled corticosteroids were compared to either placebo or standard treatment in patients with cystic fibrosis. Trials employing random treatment allocation and those using quasi-random allocation methods such as alternate allocation to treatment and control group were included. DATA COLLECTION AND ANALYSIS: The following outcomes were assessed: objective measures of lung function, respiratory exacerbations, use of intravenous antibiotics, hospital admissions, nutritional status, symptoms, quality of life, survival and frequency of adverse effects. Methodological quality of trials was assessed independently using established criteria by two reviewers, who also extracted relevant data independently using standard proformas. Differences were resolved by discussion. MAIN RESULTS: Nine trials were identified reporting the use of inhaled steroids in 266 subjects aged between seven and 45 years with cystic fibrosis. Methodological quality was difficult to assess from published information, specifically with respect to concealment of allocation and method used to generate random sequence. Trials were heterogeneous with respect to inclusion criteria, specifically age, severity of pulmonary involvement, clinical diagnosis of asthma and pulmonary colonisation with Pseudomonas aeruginosa. Trials also differed in type and duration of treatment. Beclomethasone was given for periods of between four and 22 weeks in four trials, budesonide for six weeks and six months respectively in two, and fluticasone for periods of between six weeks and two years in the remaining three. Measures of the volume of air breathed out on a forcible expiration (forced expiratory volumes) were reported in most trials but these data could not be combined for this review partly because reports differed in the way data were summarised and partly because some data were not included in published reports. Outcomes of potentially greater relevance to affected individuals such as nutritional status or quality of life were not reported in any trial. Survival was not reported in any trial, but this may reflect the fact that maximum duration of follow up was too short to allow this outcome to be meaningfully assessed. Adverse effects were systematically documented in only two trials. Although one trial was halted prematurely because a proportion of all those taking part had acquired chronic lung infections with Pseudomonas aeruginosa, no conclusions can be reached from this one small trial as to whether this risk is increased as PMID- 10796839 TI - Nocturnal mechanical ventilation for chronic hypoventilation in patients with neuromuscular and chest wall disorders. AB - BACKGROUND: Chronic alveolar hypoventilation is a common complication of many neuromuscular and chest wall disorders. Long term nocturnal mechanical ventilation is used to treat an increasing number of patients. OBJECTIVES: To examine the efficacy of nocturnal mechanical ventilation in relieving hypoventilation related symptoms in patients with neuromuscular or chest wall disorders. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group register for randomized trials and enquiry from authors of trials and other experts in the field. SELECTION CRITERIA: Types of studies: quasi-randomized or randomized controlled trials TYPES OF PARTICIPANTS: patients with neuromuscular or chest wall disorder-related stable chronic hypoventilation of all ages and all degrees of severity. Types of interventions: any type and any mode of nocturnal mechanical ventilation. Types of outcome measures: Primary: short term and long term reversal of hypoventilation related clinical symptoms Secondary: unplanned hospital admission rate, one year mortality, short term and long term reversal of day time hypercapnia, improvement of lung function and improvement of sleep breathing disorders. DATA COLLECTION AND ANALYSIS: We identified four randomized trials. One author extracted the data and another checked them. Individual data were available from the authors of the largest study. MAIN RESULTS: The four eligible trials included a total of 51 patients. The risk difference (proportion of patients) of no improvement of hypoventilation related clinical symptoms in the short term following nocturnal mechanical ventilation was significant and favoured treatment, -0.417 (95% CI -0.639 to -0.194). However, there was significant heterogeneity across the studies (p<0.001). Similarly, the risk difference of no reversal of day time hypercapnia in the short term following nocturnal ventilation was significant and favoured treatment, -0.635 (95% CI 0.874 to -0.396). The weighted mean difference of nocturnal mean oxygen saturation percent was 5.5 (95% CI 1.5 to 9.4) more improvement in patients treated with nocturnal mechanical ventilation. For the primary and most of the secondary outcome measures there was no significant difference between nocturnal mechanical ventilation and no ventilation in the long term, except for one-year mortality. Indeed, the risk difference of death one year following implementation of nocturnal mechanical ventilation was significant and favoured treatment, 0.259 (95% CI -0.478 to -0. 041). However, there was significant heterogeneity across the studies (p<0.001). Most of the secondary outcomes were not assessed in the eligible trials. No data could be summarised for the comparisons between invasive and non-invasive mechanical ventilation, between intermittent positive pressure and negative pressure ventilation, and between volume-cycled and pressure-cycled ventilation. REVIEWER'S CONCLUSIONS: Current evidence about the therapeutic benefit of mechanical ventilation is weak, but consistent, suggesting alleviation of the symptoms of chronic hypoventilation in the short term, and in two small studies survival was prolonged. Mechanical ventilation should be offered as a therapeutic option to patients with chronic hypoventilation due to neuromuscular diseases. Further larger randomized trials are needed to confirm long term beneficial effects of nocturnal mechanical ventilation on quality of life, morbidity and mortality, to assess its cost-benefit ratio, and to compare the different types and modes of ventilation. PMID- 10796840 TI - Pharmacological interventions for non-ulcer dyspepsia. AB - BACKGROUND: The commonest cause of upper gastrointestinal symptoms is non-ulcer dyspepsia (NUD) and yet the pathophysiology of this condition has been poorly characterised and the optimum treatment is uncertain. It is estimated that pound450 million is spent on dyspepsia drugs in the UK each year. OBJECTIVES: This review aims to determine the effectiveness of six classes of drugs (antacids, histamine H2 antagonists, proton pump inhibitors, prokinetics, mucosal protecting agents and antimuscarinics) in the improvement of either the individual or global dyspepsia symptom scores and also quality of life scores patients with non-ulcer dyspepsia. SEARCH STRATEGY: Trials were located through electronic searches of the Cochrane Controlled Trials Register (CCTR), MEDLINE, EMBASE, CINAHL and SIGLE, using appropriate subject headings and text words, searching bibliographies of retrieved articles, and through contacts with experts in the fields of dyspepsia and pharmaceutical companies. SELECTION CRITERIA: All randomised controlled trials (RCTs) comparing drugs of any of the six groups with each other or with placebo for non-ulcer dyspepsia (NUD). DATA COLLECTION AND ANALYSIS: Data were collected on dyspeptic symptom scores either individual or global symptom assessments and also quality of life scores and adverse effects. MAIN RESULTS: A total of 11775 citations were obtained. 144 trials were retrieved and 94 trials fulfilled our eligibility criteria. However, subsequent data extraction was not possible in 37 trials. Fifty trials were excluded as they did not meet our eligibility criteria. The final 57 trials were included in the final meta-analysis. Prokinetics (12 trials with dichotomous outcomes generating 829 patients; relative risk reduction [RRR] = 50%; 95% confidence intervals [CI] = 30 to 65%) and H2RAs (8 trials generating 1,125 patients; RRR = 30%; 95% CI = 4 to 48%) were significantly more effective than placebo. Proton pump inhibitors (4 trials generating 1,248 patients; RRR = 12%; 95% CI = -1 to 24%) and Bismuth salts (6 trials generating 311 patients; RRR = 40%; 95% CI = -3 to 65%) were superior to placebo but this was of marginal statistical significance. Antacids (one trial generating 109 patients; RRR = -2%; 95% CI = -36% to 24%) and sucralfate (two trials generating 246 patients; RRR = 29%; 95% CI = -40% to 64%) were not statistically significantly superior to placebo. A funnel plot suggested that the prokinetic result could be due to publication bias. The funnel plot of H2RAs did not show any evidence of publication bias but the quality of the trials was generally poor. REVIEWER'S CONCLUSIONS: There is some evidence that anti secretory therapy may be effective in NUD. The trials evaluating prokinetic therapy are difficult to interpret as the meta-analysis result could have been due to publication bias. Further research using prokinetics and anti-secretory therapy is required before any firm conclusions can be reached. The effect of these drugs is likely to be small and many patients will need to take them on a long-term basis so the therapies assessed need to be inexpensive and well tolerated. PMID- 10796841 TI - Initial management strategies for dyspepsia. AB - BACKGROUND: This review considers management strategies (combinations of initial investigation and empirical treatments) for dyspeptic patients. OBJECTIVES: To determine the effectiveness, acceptability, and cost effectiveness of the following initial management strategies for patients presenting with dyspepsia (a) initial pharmacological therapy (including endoscopy for treatment failures) (b) early endoscopy (c) testing for Helicobacter pylori and endoscope only those positive (d) H.pylori eradication therapy with or without prior testing. SEARCH STRATEGY: Trials were located through electronic searches and extensive contact with trialists. SELECTION CRITERIA: All randomised controlled trials of dyspeptic patients presenting in primary care. DATA COLLECTION AND ANALYSIS: Data was collected on dyspeptic symptoms, quality of life and use of resources. MAIN RESULTS: Ten papers reporting 12 comparisons were found. Trials comparing proton pump inhibitors (PPI) with antacids (2 trials) and H2 receptor antagonists (3 trials), and of early endoscopy compared with initial acid suppression (3 trials) were pooled. PPIs were significantly more effective than both H2RA s and antacids. Relative risks (RR) and 95% CI were, for PPI: antacid 0.72 (0.64-0.80), PPI: H2RA 0.63 (0.47-0.85). Results for other drug comparisons were either absent or inconclusive. Early endoscopy was not more effective than initial prescribing (RR 0.90 (0.77-1.04), but current studies lack power. No eligible trials of H.pylori test and endoscopy or test and eradicate were found. REVIEWER'S CONCLUSIONS: PPIs are effective in the treatment of dyspepsia in these trials which may not adequately exclude patients with gastro-oesophageal reflux disease. The relative efficacy of H2RA and PPI is uncertain. Early investigation may benefit some patients with dyspepsia. The review will be updated shortly with several large trials that have recently been completed. PMID- 10796842 TI - Antistreptococcal interventions for guttate and chronic plaque psoriasis. AB - BACKGROUND: Guttate psoriasis is a distinctive acute form of psoriasis which characteristically occurs in children and young adults. It is closely associated with preceding streptococcal sore throat or tonsillitis. Some authorities have claimed that ordinary (chronic plaque) psoriasis may also be made worse by infection at distant sites. Although many dermatologists have recommended using antibiotics for guttate psoriasis in particular, it is not clear whether they influence the course of either form of psoriasis. Some dermatologists have also recommended tonsillectomy for psoriasis in patients with recurrent streptococcal sore throat. OBJECTIVES: To assess the evidence for effectiveness of antistreptococcal interventions including antibiotics and tonsillectomy in the management of acute guttate and chronic plaque psoriasis. SEARCH STRATEGY: We searched the Cochrane Clinical Trials Register (Cochrane Library, Issue 3, 1999), Medline (1966- September 1999), Embase (1988-September 1999), the Salford Database of Psoriasis Trials (to November 1999) and the European Dermato Epidemiology Network (EDEN) Psoriasis Trials Database (to November 1999) for terms [STREPTOCOCC* or ANTIBIOTIC* or TONSIL*] and PSORIASIS using the Cochrane Skin Group search strategy. SELECTION CRITERIA: Randomised trials of one or more antistreptococcal interventions in patients with guttate or chronic plaque psoriasis. DATA COLLECTION AND ANALYSIS: Two reviewers independently examined each retrieved trial for eligibility and quality. MAIN RESULTS: The one eligible trial we identified compared the use of two oral antibiotic schedules in 20 psoriasis patients, predominantly of guttate type, who had evidence of beta haemolytic streptococcal colonisation. Either rifampicin or placebo was added to the end of a standard course of antistreptococcal antibiotic (phenoxymethylpenicillin or erythromycin). No patient in either arm of the study improved during the observation period. No randomised trials of tonsillectomy for psoriasis were identified. REVIEWER'S CONCLUSIONS: Although it is well known that guttate psoriasis may be precipitated by streptococcal infection, there is no firm evidence to support the use of antibiotics either in the management of established guttate psoriasis or in preventing the development of guttate psoriasis following streptococcal sore throat. Although both antibiotics and tonsillectomy have frequently been advocated for patients with recurrent guttate psoriasis or chronic plaque psoriasis, there is to date no good evidence that either intervention is beneficial. PMID- 10796843 TI - Opioid antagonists and adrenergic agonists for the management of opioid withdrawal. AB - BACKGROUND: Managed withdrawal, or detoxification, is not in itself a treatment for opioid dependence, but it is a required first step for many forms of longer term treatment. It may also represent the end point of an extensive period of treatment such as methadone maintenance. As such, managed withdrawal is an essential component of an effective treatment system. This review is one of a series that aims to assess the evidence as to the effectiveness of approaches to managing opioid withdrawal. OBJECTIVES: To assess the effectiveness of interventions involving the combined use of opioid antagonists an adrenergic agonists to manage the acute phase of opioid withdrawal. SEARCH STRATEGY: Multiple electronic databases, including Medline, Embase, Psychlit, Australian Medical Index and Current Contents, were searched using a strategy designed to retrieve references broadly addressing the management of opioid withdrawal. Reference lists of retrieved studies, reviews and conferences were handsearched. SELECTION CRITERIA: Studies that were included: involved administration of an opioid antagonist in combination with an alpha2 adrenergic agonist; had modification of the signs and symptoms of withdrawal as the aim of the intervention; involved participants who had been diagnosed as primarily opioid dependent and were undergoing clinically managed withdrawal; had as their primary focus the acute phase of withdrawal; reported detail of the type and dose of drugs used and the characteristics of study participants; reported information on the nature of withdrawal symptoms experienced, the occurrence of side effects OR rates of completion of withdrawal; and were randomized or quasi-randomized controlled clinical trials or prospective controlled cohort studies comparing the combination of opioid antagonists and adrenergic antagonists with another form of treatment. (The findings of prospective single group studies or case series, and controlled studies without a comparison treatment modality were considered in the narrative component of the review without being identified as included studies). DATA COLLECTION AND ANALYSIS: Potentially relevant studies were assessed for inclusion by one reviewer (LRG). Inclusion decisions were confirmed by consultation between all three reviewers. Included studies were assessed by all reviewers. One reviewer (LRG) undertook data extraction with the process confirmed by consultation between all three reviewers. Three studies compared treatment using an opioid antagonist-clonidine combination with treatment using clonidine only. This review incorporates data tables comparing maximum withdrawal scores and numbers of participants completing withdrawal for these three studies.. The capacity for data analysis is limited by differences in the assessment outcomes in the three studies and the likelihood of allocation bias in one study. Consequently, meta-analysis has not been undertaken to combine the findings of the three studies. MAIN RESULTS: Three studies (four reports) met the criteria for inclusi on in analytical components of this review. Six further studies were identified that managed withdrawal using opioid antagonists in combination with adrenergic agonists, but which did not meet the inclusion criteria (four were single group studies, 2 were controlled studies but did not include a comparison treatment modality). Findings of these studies are considered in narrative components of the review. Naltrexone was the primary opioid antagonist used to induce withdrawal. The most common approach was to administer naltrexone once a day, using an initial dose of 12.5mg, usually on day one or day two of treatment. Doses of clonidine were generally in the range of 01.-0.3mg three times a day. Five studies provided treatment on an outpatient basis, but all studies provided extended care on the day naltrexone was first administered. (ABSTRACT TRUNCATED) PMID- 10796844 TI - Carbamazepine for cocaine dependence. AB - BACKGROUND: Cocaine dependence has become a substantial public health problem, developing a significant number of medical, psychological and social problems, including the spread of infectious diseases (e.g. AIDS, hepatitis and tuberculosis), crime, violence and neonatal drug exposure. Although there is no consensus regarding how to treat cocaine dependence, effective pharmacotherapy has a potentially major role to play as part of a broader treatment milieu. The anti-convulsant carbamazepine, a tricyclic medication that is widely used to treat a variety of neurological and psychiatric disorder, has also been used for treatment of cocaine dependence, although its effectiveness has not been established. OBJECTIVES: To determine whether carbamazapine (CBZ) is effective on the treatment of cocaine dependence. SEARCH STRATEGY: Electronic searches of Cochrane Library, EMBASE, MEDLINE, PsycLIT, Biological Abstracts and LILACS; scan of reference list of relevant articles; personal communication; conference abstracts; unpublished trials from pharmaceutical industry; book chapters on treatment of cocaine dependence. SELECTION CRITERIA: The inclusion criteria for all randomised controlled trials were that they should focus on the use of carbamazepine drugs versus placebo on the treatment of cocaine dependence. Trials including patients with additional diagnosis such as opiate dependence were also eligible. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently and Odds Ratios, weighted mean difference and number needed to treat were estimated. Qualitative assessments of the methodology of eligible studies were carried out using validated checklists. The reviewers assumed that people who died or dropped out had no improvement and tested the sensitivity of the final results to this assumption. Where possible analysis was carried out according to the "intention to treat" principles. MAIN RESULTS: 5 studies were included in the review, with 455 people randomised. No differences were found regarding positive urine sample for cocaine metabolites. Scores on Spielberg State Anxiety Inventory slightly favoured carbamazepine, but didn't reach statistical significance. Dropouts were high in both groups up to 70% in the placebo group. Less dropout occurred in the Carbamazepine group (RR 0.87 95%CI 0.71-1.06). When no retention in treatment was due to side effects no differences were found. The number of participants presenting at least one side effect, reported in Kranzler (1995), was higher in the carbamazepine group (RR 4.33 95% CI 1.45-12.91). REVIEWER'S CONCLUSIONS: There is no current evidence supporting the clinical use of CBZ in the treatment of cocaine dependence. Larger randomised investigation must be considered taking into account that these time-consuming efforts should be reserved for medications showing more relevant and promising evidence. PMID- 10796845 TI - Photodynamic therapy for neovascular age-related macular degeneration. AB - BACKGROUND: In neovascular age-related macular degeneration, new vessels grow under the retina, distorting vision and leading to scarring. This is further exacerbated if the blood vessels leak. Photodynamic therapy, originally used in cancer treatment, has been investigated as a way to treat the neovascular membranes without affecting the retina. OBJECTIVES: The aim of this review is to examine the evidence for the safety and effectiveness of photodynamic therapy in the treatment of neovascular age-related macular degeneration. SEARCH STRATEGY: We searched for trials in the Cochrane Eyes and Vision Group trials register (available in the Cochrane Controlled Trials Register), the Cochrane Controlled Trials Register, Medline and Embase. We used the Science Citation Index to search for reports that cited identified relevant study reports. We contacted experts in the field for further trials information, and we searched the reference lists of identified relevant studies for further trial reports. Searches were conducted in December 1999. SELECTION CRITERIA: We included randomised trials of photodynamic therapy in people with choroidal neovascularisation due to age-related macular degeneration. DATA COLLECTION AND ANALYSIS: Two reviewers extracted the data independently. Meta analysis was not performed. MAIN RESULTS: One published trial was identified. Outcome data were available at 12 months after the first treatment. Patients received an average of 3.7 treatments. The relative risk of losing three or more lines of visual acuity at 12 months comparing the intervention with the control group was 0.72 (95% confidence interval 0.61 to 0.86). The relative risk of losing six or more lines of visual acuity at 12 months comparing the intervention with the control group was 0.62 (95% confidence interval 0.44 to 0.87). Subgroup analyses suggest that the benefits may be confined to people with no occult choroidal neovascularisation. REVIEWER'S CONCLUSIONS: Photodynamic therapy in people with classic choroidal neovascularisation due to age-related macular degeneration is effective in preventing visual loss. This evidence is drawn from a subgroup analysis of 143 participants in one trial. Outcomes and potential adverse effects of this treatment should be monitored closely. There is no evidence that photodynamic therapy is beneficial for people with evidence of occult choroidal neovascularisation. These people should be offered treatment in the context of a randomised trial. PMID- 10796846 TI - Nystatin prophylaxis and treatment in severely immunodepressed patients. AB - BACKGROUND: Nystatin is sometimes used prophylactically in patients with severe immunodeficiency or in the treatment of fungal infection in such patients, although the effect seems to be equivocal. OBJECTIVES: To study whether nystatin decreases morbidity and mortality when given prophylactically or therapeutically to patients with severe immunodeficiency. SEARCH STRATEGY: MEDLINE and The Cochrane Library using a comprehensive search strategy. Contacted industry and scanned reference lists. SELECTION CRITERIA: Randomised trials comparing nystatin with placebo, an untreated control group, fluconazole or amphotericin B. DATA COLLECTION AND ANALYSIS: Data on mortality, invasive fungal infection and colonisation were extracted by both authors independently. The outcomes were weighted by the inverse variance. A random effects model was used unless p>0.10 for the test of heterogeneity. MAIN RESULTS: We included 10 trials (1, 122 patients). The drugs were given prophylactically in eight trials and as treatment in two. Six trials were in acute leukaemia, one mainly in cancer, one in liver transplant patients, one in critically ill surgical and trauma patients, and one in AIDS patients. Nystatin had been compared with placebo in three trials and with fluconazole in seven; the dose varied from 1.5 MIE to 72 MIE daily. The effect of nystatin was similar to that of placebo on fungal colonisation (relative risk 0.85, 95% confidence interval 0. 65 to 1.13). There was no difference between fluconazole and nystatin on mortality (relative risk 0.87, 0.52 to 1.44) whereas fluconazole was more effective in preventing invasive fungal infection (relative risk 0.42, 0.16 to 1.12) and colonisation (relative risk 0.50, 0.36 to 0.71). The results were very similar if the three studies which were not performed in cancer patients were excluded. REVIEWER'S CONCLUSIONS: Nystatin cannot be recommended for prophylaxis or treatment of Candida infections in immunodepressed patients. PMID- 10796847 TI - Biopsy versus resection for malignant glioma. AB - BACKGROUND: Patients with isolated supratentorial brain tumours, presumed to be primary on imaging, have two surgical management options - biopsy or resection. Surgical opinions appear to be equally divided when considering the relative risks and benefits of these two procedures. OBJECTIVES: To estimate the clinical effectiveness of radical surgical resection compared to simple biopsy in patients with malignant glioma. SEARCH STRATEGY: Electronic database searches of COCHRANE CONTROLLED TRIALS REGISTER (including the Cochrane Cancer Network Specialised Register of Trials), MEDLINE, CANCERLIT, EMBASE, BIOSIS and SCIENCE CITATION INDEX. Hand searching the references of all identified studies; hand searching the Journal of Neuro-Oncology over the previous 10 years, including all conference abstracts; personal communication. SELECTION CRITERIA: Randomised and clinical controlled trials were included if they compared biopsy to resection, or looked at effect of extent of resection on survival, time to progression or quality of life, for malignant glioma patients of all ages. DATA COLLECTION AND ANALYSIS: Studies were to be identified, critically appraised and data extracted by the author (SEM). For dichotomous data, Peto odds ratios (OR) with 95% confidence intervals (CI) were hoped to have been estimated. Normal continuous data were to have been summated using the weighted mean difference (WMD). MAIN RESULTS: The electronic database search yielded 2100 citations. Of these, two articles were identified for possible inclusion, however both were excluded. The hand search and personal communication were similarly unproductive. No studies were included in the review and no data was synthesised. REVIEWER'S CONCLUSIONS: Given that no qualifying studies were identified and because this is an important issue, both in terms of patient risk and benefit and health economics, the author feels it important to conduct a randomised controlled trial in this subject. PMID- 10796848 TI - Interventions for early stage Hodgkin's disease in children. AB - BACKGROUND: Hodgkin's Disease is one of the most curable cancers in children, particularly at the early stages. The debate rests now more with retention of high cures and eliminating treatment sequelae. OBJECTIVES: To assess relapse free survival and overall survival rates in children with Stage l-llA Hodgkin's Disease treated with radiotherapy, chemotherapy or a combination of the two treatments. SEARCH STRATEGY: MEDLINE, EMBASE, CINAHL, Cancer-CD and the Cochrane Library were searched for randomised controlled trials. Key words included 'Hodgkin's Disease', 'Reed-Sternberg cells' and variations of 'child'. SELECTION CRITERIA: Treatments with a combination of the following therapies; involved field radiotherapy, extended field radiotherapy, anthracycline based chemotherapy regimens, or alkylating chemotherapy agents. Patients included children to 19 years of age. DATA COLLECTION AND ANALYSIS: Seven papers were identified as meeting the selection criteria. Of these four were usable, though one only covered a small section of the paper. It was not possible to combine the outcomes as they covered different treatment regimens and the numbers in one study were inaccurate. MAIN RESULTS: The results comparing IF with EF radiotherapy for children are based on one large multi-centred trial. Although the study is substantial and methodologically rigorous, the evidence for efficacy rests simply on this one trial. The results from the other three studies are not generalizable. The outcomes could not be combined as the studies assessed different treatment regimens. In addition there is little evidence to suggest that the methodological quality, particularly in terms of randomisation and simple arithmetic, is of a sufficiently robust nature in two of the studies. REVIEWER'S CONCLUSIONS: The treatment of Hodgkin's Disease Stage l -llA for children has undergone considerable evolution and development in the previous three decades with the introduction and refinement of combined modality treatments. There is now overall consensus that the prognosis for these patients following a short course of chemotherapy and low dose local radiotherapy is very good with a large percentage achieving long-term survival or relapse free survival. The main issues and problems facing practitioners in the field is the reduction and management of late effects and sequelae of treatment. The evidence for this overall agreement is based on a mass of cohort studies that have influenced changes in practice and significantly improved outcomes. However, apart from the collaborative study looking at radiotherapy alone, there is little evidence from randomised controlled trials to support this consensus. PMID- 10796849 TI - Surgical interventions for early squamous cell carcinoma of the vulva. AB - BACKGROUND: Radical surgery has been standard treatment for patients with early vulvar cancer since mid century. Survival figures are excellent, but complication rates are high. Over the last two decades, surgical treatment has become more individualised in order to decrease complications in patients with limited disease. OBJECTIVES: To determine whether the effectiveness and safety of individualised treatment is comparable with that of more extensive (non individualised) surgery. SEARCH STRATEGY: The cirteria set by the Cochrane Gynaecological Cancer Group were used. We searched Medline and Embase (last search on 16 November 1999) We used our own publication archives, based on a prospective handsearch of six leading relevant journalswhich was started in December 1986. Reference lists of identified studies, gynaecological cancer handbooks and conference abstracts were also used. SELECTION CRITERIA: Types of study: RCT's, case control and observational studies on the effectiveness of surgical treatment of vulvar cancer. TYPES OF PARTICIPANTS: patients with cT1N0M0 squamous cell carcinoma of the vulva. Types of interventions: local surgical treatment as well as regional lymph node dissection. Types of outcome measurements: overall, disease specific and disease free survival; treatment complications; quality of life issues. DATA COLLECTION AND ANALYSIS: The two reviewers independently assessed study quality and extracted data. MAIN RESULTS: Only two studies with a total of 94 participants were included in the review. Both were observational studies. None of the other eleven considered studies met the minimum criteria as set by the Cochrane Collaboration. From these two studies, it can be concluded that: 1. radical local excision is as safe as a radical vulvectomy; 2. An ipsilateral lymph node dissection is safe in patients with a well lateralised tumour, and 3. A superficial groin node dissection is not as safe as a full femoro-inguinal groin node dissection. The fourth question we intended to answer is of great clinical importance: is the triple incision technique as safe as an en bloc dissection? This question could only be answered by using some of the unselected studies. From these studies, the triple incision technique appears to be as safe as the en bloc technique. REVIEWER'S CONCLUSIONS: The available evidence regarding surgical treatment of early vulvar cancer is generally of poor quality. From the evidence with sufficient quality we conclude that radical local excision, ipsilateral lymph node dissection in lateral tumors and triple incision technique are safe treatment options for early vulvar cancer. However, superficial groin node dissection results in an excess of groin recurrences compared to a full femoro-inguinal groin node dissection. PMID- 10796850 TI - Three- or four- versus two-drug antiretroviral maintenance regimens for HIV infection. AB - BACKGROUND: Combination antiretroviral therapy administered to HIV-infected individuals has been shown to improve immunologic function and delay the progression of HIV infection. However, because patient adherence to complicated combination-therapy antiretroviral regimens is difficult and because of concerns regarding cumulative toxicity of antiretroviral drugs, regimens that utilize fewer antiretroviral agents are desirable. OBJECTIVES: To compare the use three- or four- versus two-drug antiretroviral maintenance regimens following successful induction therapy for HIV infection. SEARCH STRATEGY: The following electronic databases were searched for relevant randomized trials or reviews: 1. MEDLINE for the years 1982-1999 using the search terms human immunodeficiency virus, antiretroviral therapy, maintenance therapy, zidovudine, lamivudine, indinavir, stavudine, saquinivir, nelfinavir, didanosine, zalcitabine, ritonovir, AIDS, anti HIV agents, HIV infection and HIV seropositivity 2. AIDSLINE for the years 1982 1999 using the search terms antiretroviral therapy, maintenance therapy, zidovudine, lamidvudine, indinavir, stavudine, saquinivir, nelfinavir, didanosine, zalcitabine, ritonovir, anti-HIV agents 3. The Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effectiveness and the Cochrane Clinical Trials Register in the Cochrane Library, Issue 2, 1999. 4. The specialist register of trials maintained by the Cochrane Collaborative Review Group on HIV Infection and AIDS 5. AIDSTRIALS, a specialist registry of current and completed trials maintained by the U.S. National Library of Medicine The abstracts of relevant conferences, including the International Conferences on AIDS, the Conference on Retroviruses and Opportunistic Infections, the Infectious Disease Society of America annual meeting and the Interscience Conference on Antimicrobial Agents and Chemotherapy, as indexed by AIDSLINE, were also reviewed. All reference lists of all review articles and primary articles identified were searched. SELECTION CRITERIA: Randomized controlled trials in which HIV-infected adults who had successfully completed three- or four-drug antiretroviral induction therapy were randomized to maintenance therapy with three or four drugs or maintenance therapy with two drugs. Successful induction therapy was defined by a plasma viral load of less than 500 copies/ml. DATA COLLECTION AND ANALYSIS: Two reviewers assessed eligibility and trial quality. Attempts were made to contact the authors of the included abstract. Data on the number of patients experiencing loss of viral suppression were abstracted by two reviewers. The data were pooled, where appropriate, to yield odds ratios, using random effects models. MAIN RESULTS: Four trials were identified including three published studies and one abstract. Compared to three- or four-drug maintenance therapy, maintenance therapies including fewer drugs were associated with a higher risk of virologic failure (loss of HIV suppression to non-detectable levels). Combining the results of all four studies yielded an odds ratio of 5.55 (95% confidence interval, 3.14 - 9.80). Similar results were obtained when the one abstract was excluded (odds ratio, 5.48; 95% confidence interval, 2.82 - 10.65). Performing subgroup analyses of studies using the same induction and maintenance regimens gave similar results. Maintenance regimens of zidovudine and lamivudine compared to maintenance regimens with zidovuine, lamivudine and indinavir, were associated with significantly higher rates of virologic failure (odds ratio, 4.57; 95% confidence interval, 1.80 - 11.58). Similarly, maintenance regimens that discontinued one or more protease inhibitor after including them in induction therapy were also associated with a significantly higher risk of virologic failure (odds ratio, 6.15; 95% confidence interval, 3.40 -11.10). (ABSTRACT TRUNCATED) PMID- 10796851 TI - Zidovudine (AZT) versus AZT plus didanosine (ddI) versus AZT plus zalcitabine (ddC) in HIV infected adults. AB - BACKGROUND: Zidovudine (AZT) monotherapy was the first antiretroviral drug to be tested widely. The next two drugs to be developed were didanosine (ddI) and zalcitabine (ddC). OBJECTIVES: To assess the effects of zidovudine (AZT), zidovudine plus didanosine (ddI) and zidovudine plus zalcitabine (ddC) on HIV disease progression and survival. SEARCH STRATEGY: Investigators and pharmaceutical companies were contacted, and MEDLINE searches were supplemented by searching conference abstracts. SELECTION CRITERIA: Randomised controlled trials comparing any two of AZT plus ddI, AZT plus ddC or AZT alone in participants with or without AIDS which collected information on deaths and new AIDS events. DATA COLLECTION AND ANALYSIS: Individual patient data with, wherever possible, follow-up obtained beyond that previously published were obtained and checked for internal consistency and consistency with any published reports; any apparent discrepancies were resolved with the trialists. Time to death and to disease progression (defined as a new AIDS-defining event or prior death) were analysed on an intention to treat basis, stratified to avoid direct comparisons between participants in different trials. MAIN RESULTS: Six trials were included in the meta-analysis. During a median follow-up of 29 months, 2904 individuals progressed, of whom 1850 died. The addition of ddI to AZT delayed both progression (RR 0.74; 95% CI 0.67 to 0.82, P<0.0001) and death (RR 0.72; 95% CI 0.64 to 0.82, P<0.0001). Likewise, the addition of ddC to AZT also delayed progression (RR 0. 86; 95% CI 0.78 to 0.94, P=0.001) and death (RR 0.87; 95% CI 0.77 to 0.98, P=0.02). After 3 years the estimated percentages alive and without a new AIDS event were 53% for AZT+ddI, 49% for AZT+ddC and 44% for AZT alone; the percentages alive were 68%, 63% and 59% respectively. Five of the six trials involved randomised comparisons of AZT+ddI versus AZT+ddC: in these, the AZT+ddI regimen had greater effects on disease progression (P=0.004) and death (P=0.009). REVIEWER'S CONCLUSIONS: The use of ddI and, to a lesser extent, ddC delayed both HIV disease progression and death, at least when added to AZT. PMID- 10796853 TI - Hyperbaric oxygen for carbon monoxide poisoning. AB - BACKGROUND: Poisoning with carbon monoxide (CO) remains an important cause of accidental and intentional injury worldwide. Several unblinded nonrandomized trials have suggested that the use of hyperbaric oxygen (HBO) prevents the development of neurological sequelae. This has led to the widespread use of HBO in the management of patients with carbon monoxide poisoning. OBJECTIVES: To assess the effectiveness of hyperbaric oxygen (HBO) compared to normobaric oxygen (NBO) for the prevention of neurologic symptoms in patients with acute carbon monoxide poisoning. SEARCH STRATEGY: We searched MEDLINE (1966-present), EMBASE (1980-present), and the Controlled Trials Register of the Cochrane Collaboration, supplemented by a manual review of bibliographies of identified articles and discussion with recognized content experts. SELECTION CRITERIA: All randomized controlled trials involving non pregnant adults acutely poisoned with carbon monoxide, regardless of severity, with adequate or unclear allocation concealment were examined. Trials with a score of 3 out of 5 or higher on the validity instrument of Jadad were included in the primary analysis. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted from each trial information on the number of randomized patients, types of participants, the dose and duration of the intervention, and the prevalence of neurologic symptoms at follow-up. A pooled odds ratio (OR) for the presence of neurologic symptoms at one month follow-up was calculated using a random effects model. MAIN RESULTS: Six randomized controlled trials were identified. The trials were of varying quality. Three trials employing different doses of NBO and HBO were included in the primary analysis. The severity of CO poisoning was inconsistent between trials. At one month follow-up after treatment, symptoms possibly related to carbon monoxide poisoning were present in 81 of 237 patients (34.2%) treated with HBO, compared with 81 of 218 patients (37.2%) treated with NBO (O.R. for benefit with HBO 0.82; 95% CI 0.41-1.66). REVIEWER'S CONCLUSIONS: There is no evidence that unselected use of HBO in the treatment of acute CO poisoning reduces the frequency of neurological symptoms at one month. However, evidence from the available randomized controlled trials is insufficient to provide clear guidelines for practice. Further research is needed to better define the role of HBO, if any, in the treatment of carbon monoxide poisoning. This research question is ideally suited to a multicentre, randomized, double-blind controlled trial. PMID- 10796852 TI - Immediate versus deferred zidovudine (AZT) in asymptomatic or mildly symptomatic HIV infected adults. AB - BACKGROUND: Zidovudine (AZT) monotherapy was the first antiretroviral drug to be tested widely. Subsequent trials in asymptomatic or early symptomatic HIV infection indicated short-term delays in disease progression with AZT, but not improved survival. OBJECTIVES: To assess the effects of immediate versus deferred zidovudine (AZT) on HIV disease progression and survival. SEARCH STRATEGY: Investigators and pharmaceutical companies were contacted, and MEDLINE searches were supplemented by searching conference abstracts. SELECTION CRITERIA: Randomised controlled trials comparing immediate versus deferred AZT in participants without AIDS which prospectively collected deaths and new AIDS events. DATA COLLECTION AND ANALYSIS: Individual patient data with, wherever possible, follow-up obtained beyond that previously published was obtained and checked for internal consistency and consistency with any published reports; any apparent discrepancies were resolved with the trialists. Time to death and to disease progression (defined as a new AIDS-defining event or prior death) were analysed on an intention to treat basis, stratified to avoid direct comparisons between participants in different trials. MAIN RESULTS: Nine trials were included in the meta-analysis. During a median follow-up of 50 months, 1908 individuals developed disease progression, of whom 1351 died. In the deferred group, 61% started antiretroviral therapy (median time to therapy 28 months, which was AZT monotherapy in 94%). During the first year of follow-up immediate AZT halved the rate of disease progression (P<0.0001), increasing the probability of AIDS-free survival at one year from 96% to 98%, but this early benefit did not persist: after 6 years AIDS-free survival was 54% in both groups, and at no time was there any difference in overall survival, which at 6 years was 64% with immediate and 65% with deferred AZT (rate ratio [RR] 1.04, 95% confidence interval [CI] 0. 94 to 1.15). REVIEWER'S CONCLUSIONS: Although immediate use of AZT halved disease progression during the first year, this effect was not sustained, and there was no improvement in survival in the short or long term. PMID- 10796854 TI - Premedication for anxiety in adult day surgery. AB - BACKGROUND: Surgery is increasingly performed on a day-case basis. Many patients are anxious pre-operatively and might benefit from pharmacological anxiolysis. Drugs are sometimes not used, however, for fear of delaying discharge from hospital. OBJECTIVES: To asses the effect of anxiolytic premedication on time to discharge in adult patients undergoing day case surgery under general anaesthesia. SEARCH STRATEGY: Trials were identified by computerised searches of the Cochrane Controlled Trials Register, MEDLINE, EMBASE, by checking the reference lists of trials and review articles, by hand-searching three main anaesthesia journals and by contacting five researchers active in the field and the Product Information departments of the manufacturers of five commonly used premedicants. SELECTION CRITERIA: All randomised controlled trials comparing an anxiolytic drug(s) with placebo before general anaesthesia in adult day case surgical patients. DATA COLLECTION AND ANALYSIS: We collected data on anaesthetic drugs used, results of tests of psychomotor function where these were used to assess residual effect of premedication, and on times from end of anaesthesia to ability to walk unaided or readiness for discharge from hospital. Formal statistical synthesis of individual trials was not performed in view of the variety of drugs studied. MAIN RESULTS: Searching identified twenty-nine reports; fourteen studies, with data from a total of 1263 patients, were considered eligible for analysis. Only two studies specifically addressed the discharge question; both found no delay in premedicated patients. Three other studies used clinical criteria to assess fitness for discharge, though times were not given. Again, there was no difference from placebo. Four studies used both clinical measures and tests of psychomotor function as tests of recovery from anaesthesia. In none of these studies did the premedication appear to delay discharge, although performance on tests of psychomotor function was sometimes still impaired. Of the four studies which used tests of psychomotor function to assess recovery, three showed impaired recovery (after midazolam 7.5mg, midazolam 15mg or diazepam 15mg) which might possibly interfere with discharge from hospital. REVIEWER'S CONCLUSIONS: We have found no evidence of a difference in time to discharge from hospital in patients who received anxiolytic premedication. However, in view of the age and variety of anaesthetic techniques used, inferences for current day-case practice should be made with caution. PMID- 10796855 TI - Single dose oral aspirin for acute pain. AB - BACKGROUND: Aspirin has been known to be an effective analgesic for many years and is commonly used throughout the world for many different pain conditions. It is important for both prescribers and patients to have the best possible information about the efficacy and safety of analgesics, and this need is reflected in patient surveys which show that postoperative pain is often poorly managed. We also need to benchmark relative efficacy and safety of current analgesics so that we can compare them with new analgesics. OBJECTIVES: To quantitatively assess the analgesic efficacy and adverse effects of a single-dose of aspirin in acute pain of moderate to severe intensity. SEARCH STRATEGY: Randomised trials were identified by searching Medline (1966 to March 1998), Embase (1980 to January 1998), the Cochrane Library (Issue 1,1998) and the Oxford Pain Relief Database (1950 to 1994). SELECTION CRITERIA: The inclusion criteria used were: full journal publication, postoperative pain or a mixture of postoperative and acute trauma pain, oral administration, adult patients, baseline pain of moderate to severe intensity, double-blind design, and random allocation to treatment groups which compared aspirin with placebo. DATA COLLECTION AND ANALYSIS: Summed pain relief or pain intensity difference over four to six hours was extracted, and converted into dichotomous information yielding the number of patients with at least 50% pain relief. This was then used to calculate the relative benefit and the number-needed-to-treat (NNT) for one patient to achieve at least 50% pain relief. MAIN RESULTS: Seventy-two randomised single-dose trials met our inclusion criteria, with 3253 patients given aspirin, and 3297 placebo. Significant benefit of aspirin over placebo was shown for aspirin 600/650 mg, 1000 mg and 1200 mg, NNTs for at least 50% pain relief of 4.4 (4.0 to 4.9), 4.0 (3.2 to 5.4) and 2.4 (1.9 to 3.2) respectively. Single-dose aspirin 600/650 mg produced significantly more drowsiness and gastric irritation than placebo, with a number-needed-to-harm (NNH) of 28 (19 to 52) and 38 (22 to 174) respectively. Type of pain model, pain measurement, sample size, quality of study design, and study duration had no significant impact on the results. REVIEWER'S CONCLUSIONS: Aspirin is an effective analgesic for acute pain of moderate to severe intensity with a clear dose-response. Drowsiness and gastric irritation were seen as significant adverse effects even though the studies were single-dose. The pain relief achieved with aspirin was very similar milligram for milligram to that seen with paracetamol. PMID- 10796856 TI - Minocycline for acne vulgaris: efficacy and safety. AB - BACKGROUND: Minocycline is a tetracycline antibiotic that is commonly used in the treatment of moderate to severe acne vulgaris. Although it is more convenient for patients to take than first-generation tetracyclines, as it only needs to be taken once or twice a day and can be taken with food, it is more expensive. Concerns have also been expressed over its safety following the deaths of two patients taking the drug. There is a lack of consensus among dermatologists over the relative risks and benefits of minocycline. As most acne prescribing is undertaken by general practitioners, it is important that guidelines issued to them are based on the best available evidence rather than personal judgements. OBJECTIVES: To collate and evaluate the evidence on the clinical efficacy of minocycline in the treatment of inflammatory acne vulgaris. Specific objectives were to compare the efficacy of minocycline with other drug treatments for acne and to collate information on the incidence of adverse drug reactions. SEARCH STRATEGY: Randomised controlled trials (RCTs) of minocycline for acne vulgaris were identified by searching the following electronic databases; MEDLINE, EMBASE, Biosis, Biological Abstracts, International Pharmaceutical Abstracts, Cochrane Skin Group's Trial Register, Theses Online, BIDS ISI Science Citation Index and Bids Index to Scientific and Technical Proceedings. Other strategies used were scanning the references of articles retrieved, hand-searching of major dermatology journals and personal communication with trialists and drug companies. SELECTION CRITERIA: To be eligible for the review, studies had to be RCTs comparing the efficacy of minocycline at any dose to active or placebo control, in subjects with inflammatory acne vulgaris. Diagnoses of papulo pustular, polymorphic and nodular acne were also accepted. Trials were not excluded on the basis of language. DATA COLLECTION AND ANALYSIS: 27 randomised controlled trials met the inclusion criteria and were included in this review. The comparators used were placebo (2 studies), oxytetracycline (1), tetracycline (6), doxycycline (7), lymecycline (2), topical clindamycin (3), topical erythromycin/zinc (1), cyproterone acetate/ ethinyloestradiol (1), oral isotretinoin (2), topical fusidic acid (1) and there was one dose response study. One study is ongoing and it remains to be clarified whether one further study is a RCT. Major outcome measures used in the trials included lesion counts, acne grades/severity scores, doctors' and patients' global assessments, adverse drug reactions and drop out rates. The quality of each study was assessed independently by two assessors and an effect size calculated where possible. MAIN RESULTS: The trials were generally small and of poor quality and in many cases the published reports were inadequate for our purpose. Pooling of the studies was not attempted due to the lack of common outcome measures and endpoints and the unavailability of some primary data. Although minocycline was shown to be an effective treatment for acne vulgaris, in only two studies was it found to be superior to other tetracyclines. Both of these were conducted under open conditions and had serious methodological problems. A third study showed it to be more effective than 2% fusidic acid, applied topically, against inflammatory lesions in mild to moderate acne. Differences in the way adverse drug reactions were identified could have accounted for the wide variation between studies in numbers of events reported. This meant that no overall evaluation could be made of incidence rates of adverse events associated with minocycline therapy. No RCT evidence was found to support the benefits of minocycline in acne resistant to other therapies and the dose response has only been evaluated up to eight weeks of therapy. (ABSTRACT TRUNCATED) PMID- 10796857 TI - Short-term treatment with proton pump inhibitors, H2-receptor antagonists and prokinetics for gastro-oesophageal reflux disease-like symptoms and endoscopy negative reflux disease. AB - BACKGROUND: Heartburn affects 25% of the adult population on a monthly basis and represents the core symptom of gastro-oesophageal reflux disease (GORD). Treatment is readily available and puts a large demand on healthcare budgets. Research in the past has focused largely on the treatment of oesophagitis. A majority of GORD patients show no endoscopic abnormalities and in daily practice most patients are treated empirically. OBJECTIVES: Summarise, quantify and compare the efficacy of the short-term use of proton pump inhibitors (PPI), H2 receptor antagonists (H2RA) and prokinetics in the empirical treatment of GORD and the treatment of endoscopy negative reflux disease (ENRD). SEARCH STRATEGY: Electronic searches were performed of the Cochrane Controlled Trials Register, MEDLINE and EMBASE. Bibliographies were screened. SELECTION CRITERIA: Included were randomised controlled trials focussing on symptomatic outcome after short term treatment for GORD using proton pump inhibitors, H2-receptor antagonists or prokinetic agents. Participants had to be classifiable in the empirical treatment group (no endoscopy used in treatment allocation) or in the endoscopy negative reflux disease group (no endoscopic signs of erosive oesophagitis). DATA COLLECTION AND ANALYSIS: Data from included trials were extracted by two reviewers independently. The impact of interventions was expressed as relative risks (RR) together with 95% confidence intervals (95% CI). Meta-analysis was only performed if there were sufficient trials of similar comparisons reporting the same outcomes. Relative risks were combined for binary outcomes. MAIN RESULTS: Twenty-one trials were included: eleven in the empirical treatment group, seven in the ENRD group and three in both. In empirical treatment of GORD the RR for heartburn remission in placebo-controlled trials for PPI was 0.35 (1 trial, 95% CI 0.26 to 0.46), for H2RAs 0.77 (2 trials, 95% CI 0.60 to 0.99) and for prokinetics 0.86 (1 trial, 95% CI 0.73 to 1.01). In direct comparison PPIs were significantly (p<0.05) more effective than H2RAs (3 trials, RR 0.67, 95% CI 0.57 to 0.80) and prokinetic's (2 trials, RR 0.53, 95% CI 0.32 to 0.87). In treatment of ENRD, RR for heartburn remission for PPI versus placebo was 0.68 (4 trials, 95% CI 0.53 to 0.88) and for H2RA versus placebo was 0.84 (2 trials, 95% CI 0.74 to 0.95). The RR for PPI versus H2RA was 0.69 (2 trials, 95% CI 0.39 to 1.20) and versus prokinetic 0.72 (1 trial, 95% CI 0.56 to 0.92). REVIEWER'S CONCLUSIONS: The findings in this review suggest that antisecretory drugs are effective in the empirical treatment of complaints likely to originate from GORD and in treatment of ENRD and furthermore that PPIs are superior to H2RAs in empirical treatment of typical GORD symptoms. PMID- 10796858 TI - Eradication of helicobacter pylori for non-ulcer dyspepsia. AB - BACKGROUND: Helicobacter pylori (H pylori) is the main cause of peptic ulcer disease. The role of H pylori in non-ulcer dyspepsia is less clear. OBJECTIVES: To determine the effect of H pylori eradication on dyspepsia symptoms and quality of life scores in patients with non-ulcer dyspepsia. SEARCH STRATEGY: Trials were identified through electronic searches of the Cochrane Controlled Trials Register (CCTR), MEDLINE, EMBASE, CINAHL and SIGLE, using appropriate subject headings and keywords, searching bibliographies of retrieved articles, and through contacts with experts in the fields of dyspepsia and with pharmaceutical companies. SELECTION CRITERIA: All parallel group randomised controlled trials (RCTs) comparing drugs to eradicate H pylori with placebo or other drugs known not to eradicate H pylori for patients with non-ulcer dyspepsia. DATA COLLECTION AND ANALYSIS: Data were collected on individual and global dyspeptic symptom scores, quality of life measures and adverse effects. Dyspepsia outcomes were dichotomised into minimal/resolved versus same/worse symptoms. MAIN RESULTS: Seven randomised controlled trials were included in the systematic review. Five trials compared proton pump inhibitor dual or triple therapy with a proton pump inhibitor + placebo antibiotics, and evaluated dyspepsia at 6-12 months in 1,385 patients. H pylori eradication was significantly superior to placebo in treating non ulcer dyspepsia (relative risk reduction = 7%; 95% CI = 1% to 12%; p=0.02) and there was no significant heterogeneity between the studies. The number needed to treat to cure one case of dyspepsia = 19 (95% CI = 11 to 132). A further two trials compared Bismuth based H pylori eradication with an alternative pharmacological agent. These trials were smaller and had a shorter follow-up but suggested H pylori eradication was more effective than either H2 receptor antagonists or sucralfate in treating non-ulcer dyspepsia. REVIEWER'S CONCLUSIONS: H pylori eradication may be an effective therapy for H pylori positive non-ulcer dyspepsia. This result is not robust and further evidence on the efficacy of H pylori eradication in non ulcer dyspepsia would be helpful. The effect is modest and economic models would help establish whether this approach is cost-effective. This review will be updated as results from other ongoing trials (Malfertheiner 2000) are made available. PMID- 10796859 TI - Biofeedback and/or sphincter exercises for the treatment of faecal incontinence in adults. AB - BACKGROUND: Faecal incontinence is a particularly embarrassing and distressing condition with significant medical, social and economic implications. Sphincter exercises and biofeedback therapy have been used to treat the symptoms of people with faecal incontinence. However, standards of treatment are still lacking and the magnitude of alleged benefits has yet to be established. OBJECTIVES: To determine the effects of biofeedback and/or anal sphincter exercises/pelvic floor muscle training for the treatment of faecal incontinence in adults. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and all reference lists of relevant articles up to November 1999. Date of the most recent searches: November 1999. SELECTION CRITERIA: All randomised or quasi-randomised trials evaluating biofeedback and/or anal sphincter exercises in adults with faecal incontinence. DATA COLLECTION AND ANALYSIS: Three reviewers assessed the methodological quality of eligible trials and two reviewers independently extracted data from included trials. A wide range of outcome measures were considered. MAIN RESULTS: Only five eligible studies were identified with a total of 109 participants. In the majority of trials methodological quality was poor or uncertain. All trials were small and employed a limited range of outcome measures. Follow-up information was not consistently reported amongst trials. Only two trials provided data in a form suitable for statistical analyses. There are suggestions that rectal volume discrimination training improves continence more than sham training and that anal biofeedback combined with exercises and electrical stimulation provides more short-term benefits than vaginal biofeedback and exercises for women with obstetric-related faecal incontinence. Further conclusions are not warranted from the available data. REVIEWER'S CONCLUSIONS: The limited number of identified trials together with their methodological weaknesses do not allow a reliable assessment of the possible role of sphincter exercises and biofeedback therapy in the management of people with faecal incontinence. There is a suggestions that some elements of biofeedback therapy and sphincter exercises may have a therapeutic effect, but this is not certain. Larger well-designed trials are needed to enable safe conclusions. PMID- 10796860 TI - Desmopressin for nocturnal enuresis in children. AB - BACKGROUND: Enuresis (bedwetting) is a socially disruptive and stressful condition which affects around 15-20% of five year olds, and up to 2% of young adults. Although there is a high rate of spontaneous remission, the social, emotional and psychological costs to the children can be great. OBJECTIVES: To assess the effects of desmopressin on nocturnal enuresis in children, and to compare desmopressin with other interventions. SEARCH STRATEGY: The following electronic databases were searched: MEDLINE to June 1997; AMED; ASSIA; BIDS; BIOSIS Previews (1985-1996); CINAHL; DHSS Data; EMBASE (1974 to June 1997); PsycLIT and SIGLE. Organisations, manufacturers, researchers and health professionals concerned with enuresis were contacted for information. The reference sections of obtained studies were also checked for further trials. Date of the most recent search: July 1997. SELECTION CRITERIA: All randomised trials of desmopressin for nocturnal enuresis in children were included in the review. Trials were eligible for inclusion if: children were randomised to receive desmopressin compared with placebo, other drugs or other conservative interventions for nocturnal bedwetting; participants with organic causes for their bedwetting were excluded; and baseline assessments of the level of bedwetting were reported. Trials focused solely on daytime wetting were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the quality of the eligible trials, and extracted data. MAIN RESULTS: Twenty one randomised trials involving 948 children treated with desmopressin, met the inclusion criteria. The quality of many of the trials was poor. Desmopressin was compared with a tricyclic drug in two trials, and with alarms in one. Desmopressin was effective in reducing bedwetting in a variety of doses and forms. Each dose of desmopressin reduced bedwetting by at least one night per week during treatment (eg 20microg: 1.56 fewer wet nights per week, 95% CI -1.94 to -1.19). Participants on desmopressin were 4.6 times more likely to achieve 14 consecutive dry nights (95% CI 1.38 to 15.02) compared with placebo. However, there was no difference after treatment was finished. There was no apparent dose-related effect of desmopressin, but the evidence was limited. Data which compared oral and nasal administration were too few to be conclusive. Desmopressin and imipramine (a tricyclic drug) were equally effective in one small trial. Amitriptyline (another tricyclic) was not consistently better than desmopressin either alone or when used as a supplement. In a single trial, desmopressin was initially superior to using an alarm in reducing the number of wet nights per week: WMD -1.7 (95% CI: -2.96 to -0.45), but this result was not sustained; after three months of treatment, patients using the alarm had 1.4 fewer wet nights per week than with desmopressin: (95% CI: 0.14 to 2.65). Participants receiving the alarm intervention were also nine times less likely to relapse than those given desmopressin: RR 9.2 (95% CI: 1.28 to 65.9). Combining alarm and drug therapy was found to be superior to alarm treatment alone. The addition of desmopressin to an alarm schedule resulted in one less wet night per week: (95% CI: -1.55 to -0.45). REVIEWER'S CONCLUSIONS: Desmopressin rapidly reduced the number of wet nights per week, but there was some evidence that this was not sustained after treatment stopped. Comparison with alternative treatments suggested that desmopressin and tricyclics had similar clinical effects, but that alarms produced more sustained benefits. However, based on the available evidence, these conclusions can only be tentative. There was some evidence of minor side effects of desmopressin in the included trials, such as nasal irritation and nose bleeds. However, the risk of water intoxication associated with over-drinking before bedtime has been reported. Patients and their families need to be warned of potential adverse effects and advise PMID- 10796861 TI - Prompted voiding for the management of urinary incontinence in adults. AB - BACKGROUND: Prompted voiding is a behavioural therapy used mainly in North American nursing homes. It aims to improve bladder control for people with or without dementia using verbal prompts and positive reinforcement. OBJECTIVES: To assess the effects of prompted voiding for the management of urinary incontinence in adults. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register (to February 2000) and reference lists of relevant articles. We contacted investigators in the field to locate extra studies. Date of the most recent searches: February 2000. SELECTION CRITERIA: All randomised or quasi randomised trials which addressed prompted voiding for the management of urinary incontinence. The trials included adult men and women, with or without cognitive impairment, diagnosed as having urinary incontinence as identified by the trialists, either by symptom classification or by urodynamic investigation. DATA COLLECTION AND ANALYSIS: The identified reports were assessed for eligibility. Two reviewers independently reviewed the selected studies for methodological quality. Data describing six pre-specified outcomes were extracted independently by each reviewer and consensus reached when there was disagreement. Trial investigators were consulted when clarification or further detail was required. A third reviewer was recruited to proof read the review at different stages. MAIN RESULTS: Five trials were included in the review. These involved 355 elderly people, most of whom were women. One other trial was excluded because no relevant outcome data were reported, and one trial is awaiting assessment. Prompted voiding was compared with no prompted voiding in four trials. The limited evidence suggested that prompted voiding increased self-initiated voiding and decreased incontinent episodes in the short-term. There was no evidence about long-term effects. A single small trial suggested that adding the muscle relaxant, Oxybutinin, reduced the number of incontinent episodes in the short term: This study used a cross-over design and so did not address long-term effects. REVIEWER'S CONCLUSIONS: There was insufficient evidence to reach firm conclusions for practice. There was suggestive, although inconclusive, evidence of short-term benefit from prompted voiding and from adding the muscle relaxant, Oxybutinin to prompted voiding. PMID- 10796862 TI - Weighted vaginal cones for urinary incontinence. AB - BACKGROUND: Pelvic floor muscle training has long been the most common form of conservative treatment for stress urinary incontinence. Many people have trouble identifying their pelvic floor, and thus difficulty training it. One aid to help with training the pelvic floor in women is weighted vaginal cones. These are inserted into the vagina and the pelvic floor is contracted to prevent them slipping out. OBJECTIVES: To evaluate the effects of weighted vaginal cones in the treatment of female stress urinary incontinence. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register (to December 1999), MEDLINE (January 1966 to November 1999), EMBASE (January 1988 to November 1999) and reference lists of relevant articles. Date of the most recent searches: December 1999. SELECTION CRITERIA: The studies had to be randomised or quasi-randomised controlled trials. One arm of the study had to be the use of weighted vaginal cones. The women studied had to have stress urinary incontinence. DATA COLLECTION AND ANALYSIS: Reports were assessed by all three reviewers to see if they complied with the inclusion/exclusion criteria. Data was abstracted by one reviewer and cross checked by the others. Included data were processed as described in the Cochrane Handbook. MAIN RESULTS: Ten studies fitted the inclusion and exclusion criteria. Four of these have only been published as abstracts. All of the studies were small and in many the quality was hard to judge. The studies tested seven comparisons. There was variation between studies in the ways cones were used and in the ways comparison treatments were given. Cones alone were compared with control, pelvic floor muscle training (PFMT), electrostimulation, PFMT plus cones, and electrostimulation plus PFMT. Cones in conjunction with PFMT were compared with PFMT alone and electrostimulation. Two of the studies recruited women with symptoms of incontinence while the others required that the women had urodynamically proven genuine stress incontinence. Outcome measures differed between studies, making the results difficult to combine. Some studies reported high drop out rates with some of these being related to treatment with cones. Overall the dropout rate was not much different to that in the comparison treatments. Cones were better than control treatment (cure or improvement RR 0. 38; 95% CI 0.23 to 0.63), and similar to PFMT (RR 1.23; 95% CI 0.78 to 1.94) and electrostimulation (RR 1.35; 95% CI 0.82 to 2.21). Cones plus PFMT was no different to either cones alone or PFMT alone. These results held for both subjective and objective outcomes, but the confidence intervals were wide. REVIEWER'S CONCLUSIONS: This review provides limited evidence that weighted vaginal cones are beneficial in women with stress urinary incontinence. The limited data suggest that they are better than control treatments but similar to PFMT and electrostimulation. This conclusion must remain tentative until further larger high quality studies are carried out using similar relevant outcome measures. Some women treated with cones stop treatment early so cones may only be useful for those who find them acceptable. PMID- 10796863 TI - Tricyclic and related drugs for nocturnal enuresis in children. AB - BACKGROUND: Enuresis (bedwetting) is a socially unacceptable and stressful condition which affects around 15-20% of five year olds, and up to 2% of young adults. Although there is a high rate of spontaneous remission, the social, emotional and psychological costs to the children can be great. OBJECTIVES: To assess the effects of tricyclic and related drugs on nocturnal enuresis in children, and to compare them with other interventions. SEARCH STRATEGY: The following electronic databases were searched: MEDLINE to June 1997; AMED; ASSIA; BIDS; BIOSIS Previews (1985-1996); CINAHL; DHSS Data; EMBASE (1974 to June 1997); PsycLIT and SIGLE. Organisations, manufacturers, researchers and health professionals concerned with enuresis were contacted for information. The reference sections of obtained studies were also checked for further trials. Date of the most recent search: July 1997. SELECTION CRITERIA: All randomised trials of tricyclic and related drugs for nocturnal enuresis in children were included in the review. Trials were eligible for inclusion if: children were randomised to receive tricyclics compared with placebo, other drugs or other conservative interventions for nocturnal bedwetting; participants with organic causes for their bedwetting were excluded; and baseline assessments of the level of bedwetting were provided. Trials focused solely on daytime wetting were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the quality of the eligible trials, and extracted data. MAIN RESULTS: Twenty two randomised trials, involving 1100 children treated with tricyclic or related drugs, met the inclusion criteria. The quality of many of the trials was poor. Only single trials compared tricyclic or related drugs with each other, other drugs, desmopressin, alarms or other behavioural interventions. Treatment with tricyclic drugs (such as imipramine, amitriptyline, viloxazine, clomipramine and desipramine but not mianserin) were associated with a reduction of about one wet night per week while on treatment (eg using imipramine, WMD -0.99, 95% CI -1.27 to -0.71). Children were almost five times more likely to achieve 14 dry nights with the drugs (eg using imipramine, RR = 4.99, 95% CI 2.4 to 10.40). Desmopressin and tricyclics appeared equally effective while on treatment, but this effect was not sustained after treatment stopped. There was no detectable difference between imipramine and alarms while on treatment, but afterwards those who had used alarms had one fewer wet night per week (WMD 1.03, 95% CI 0. 19 to 1.87). REVIEWER'S CONCLUSIONS: Treatment with tricyclic drugs (imipramine, amitriptyline, viloxazine, clomipramine and desipramine but not mianserin) was associated with a reduction of about one wet night per week while on treatment, but long term effectiveness is unknown. Desmopressin and tricyclics appeared equally effective while on treatment, but this effect was not sustained after treatment stopped. Alarms may be more effective in the long term. Comparisons between drug and behavioural treatments are needed, and should include relapse rates after treatment is finished. PMID- 10796864 TI - Progestagens and anti-progestagens for pain associated with endometriosis. AB - BACKGROUND: Endometriosis is a gynaecological condition that presents either with the problem of infertility or with painful symptoms. The clinical observation of an apparent resolution of symptoms during pregnancy gave rise to the concept of treating patients with a pseudo-pregnancy regime. Initially combinations of high dose oestrogens and progestagens were used but this was subsequently replaced by progestogens alone. More recently progestogens of both progestagens and anti progestagens in the treatment of symptomatiprogestogenssis OBJECTIVES: To determine the effectiveness of both the progestagens and anti-progestagens in the treatment of painful symptoms ascribed to the diagnosis of endometriosis. SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility Group was utilised to identify all publications which described or might have described randomised trials of any progestagen or any anti-progestagen in the treatment of symptomatic endometriosis. SELECTION CRITERIA: Trials were included if they were randomised and considered the effectiveness of either a progestagen or an anti progestagen in the treatment of painful symptoms associated with endometriosis. DATA COLLECTION AND ANALYSIS: Seven studies were considered to be appropriate for inclusion in this review. Only three studies evaluating progestagens were included (comparison with placebo, danazol and oral contraceptive plus danazol). All other studies compared the anti-progestagen, gestrinone, with other medical therapies. MAIN RESULTS: Progestagens appear to be an effective therapy for the painful symptoms associated with endometriosis. Gestrinone is as effective as other established medical therapies (danazol and GnRH analogues). REVIEWER'S CONCLUSIONS: The limited available data suggests that both continuous progestagens and anti-progestagens are effective therapies in the treatment of painful symptoms associated with endometriosis. Progestagens given in the luteal phase are not effective. These conclusions should be accepted cautiously due to a lack of data. PMID- 10796865 TI - Progesterone/progestogen releasing intrauterine systems versus either placebo or any other medication for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is an important cause of ill health in women and it accounts for 12% of all gynaecology referrals in the UK. Heavy menstrual bleeding is clinically defined as greater than, or equal to, 80mls blood loss per menstrual cycle but women may complain of excessive bleeding when their blood loss is less than 80ml. Hysterectomy is often used to treat women with this complaint but medical therapy may be a successful alternative. The intrauterine coil device was originally developed as a contraceptive but the addition of uterine relaxing hormones, or progestogens, to these devices resulted in a large reduction in menstrual blood loss. Case studies of 2 types of progesterone/progestogen releasing systems, Progestasert and Mirena, report reductions of up to 90% and dysmenorrhoea may be improved. Insertion, however, may be regarded as invasive by some women affecting its acceptability as a treatment and frequent intermenstrual bleeding and spotting is likely during the first few months. OBJECTIVES: To determine the effectiveness and acceptability of progesterone/progestogen-releasing intrauterine devices in achieving a reduction in heavy menstrual bleeding. SEARCH STRATEGY: All studies which might describe randomised controlled trials of progesterone/progestagen-releasing intrauterine devices for the treatment of heavy menstrual bleeding were obtained by electronic searches of the MEDLINE 1966-1999, EMBASE 1980-1999 databases and the Cochrane Library. Companies producing progestogen releasing intrauterine devices and experts in the field were contacted for information on published and unpublished trials. SELECTION CRITERIA: Randomised controlled trials in women of reproductive age treated with progesterone/progestogen-releasing intrauterine devices versus no treatment, placebo, or other medical or surgical therapy for heavy menstrual bleeding within either the primary care, family planning or specialist clinic setting were eligible for inclusion. Women with postmenopausal bleeding, intermenstrual or irregular bleeding, or pathological causes of heavy menstrual bleeding were excluded. DATA COLLECTION AND ANALYSIS: Potential trials were independently assessed by three reviewers and five trials met the criteria for inclusion in the review. The reviewers extracted the data independently and data were pooled where appropriate. Odds ratios for dichtomous outcomes and weighted mean differences for continuous outcomes were estimated from the data. The primary outcome was reduction in menstrual blood loss but incidence of side effects, changes in quality of life and satisfaction and acceptability measures were also assessed. MAIN RESULTS: Progesterone/progestogen-releasing intrauterine systems have not been compared to placebo or no treatment. Progestasert has been compared to a number of different medical therapies in one small study but no conclusions can be made about effectiveness. The levonorgestrel-releasing intrauterine device (LNG IUS) has been compared to oral cyclical norethisterone (NET) administered on days 5-26 in one trial and was significantly more effective although there was a large reduction from baseline in both groups and these differences were not perceived by the women undergoing the treatment. Some side effects were more common in the LNG IUS group but a significantly greater proportion of women in this group were satisfied and willing to continue with their treatment. In one trial of women awaiting hysterectomy where the LNG IUS was compared with a control group taking their existing medical therapy, a higher proportion of the women in the former group cancelled their planned surgery after 6 months of treatment. The levonorgestrel-releasing intrauterine device has been compared to a surgical procedure (transcervical resection of the endometrium (TCRE)) in two trials. (ABSTRACT TRUNCATED) PMID- 10796866 TI - Reduced or modified dietary fat for prevention of cardiovascular disease. AB - BACKGROUND: Reduction or modification of dietary fat can improve total cholesterol levels, but may also have a variety of effects, both positive and negative, on other cardiovascular risk factors. OBJECTIVES: The aim of this systematic review was to assess the effect of reduction or modification of dietary fats on total and cardiovascular mortality and cardiovascular morbidity over at least 6 months, using all available randomized clinical trials. SEARCH STRATEGY: The Cochrane Library, MEDLINE, EMBASE, CAB Abstracts, CVRCT registry and related Cochrane Groups' trial registers were searched through spring 1998, SIGLE to January 1999. Trials known to experts in the field and biographies were included through May 1999. SELECTION CRITERIA: Trials fulfilled the following criteria: 1) randomized with appropriate control group, 2) intention to reduce or modify fat or cholesterol intake (excluding exclusively omega-3 fat interventions), 3) not multi factorial, 4) healthy adult humans, 5) intervention at least six months, 6) mortality or cardiovascular morbidity data available. Inclusion decisions were duplicated, disagreement resolved by discussion or a third party. DATA COLLECTION AND ANALYSIS: Rate data were extracted by two independent reviewers and meta-analysis performed using random effects methodology. Meta-regression and funnel plots were used. MAIN RESULTS: Twenty seven studies were included (40 intervention arms, 30,901 person-years). There was no significant effect on total mortality (rate ratio 0.98, 95% CI 0.86 to 1.12), a trend towards protection form cardiovascular mortality (rate ratio 0.91, 95% CI 0. 77 to 1.07), and significant protection from cardiovascular events (rate ratio 0.84, 95% CI 0.72 to 0.99). The latter became non-significant on sensitivity analysis. Trials where participants were involved for more than 2 years showed significant reductions in the rate of cardiovascular events and a suggestion of protection from total mortality. The degree of protection from cardiovascular events appeared similar in high and low risk groups, but was statistically significant only in the former. REVIEWER'S CONCLUSIONS: The findings are suggestive of a small but potentially important reduction in cardiovascular risk in trials longer than two years. Lifestyle advice to all those at high risk of cardiovascular disease (especially where statins are unavailable or rationed), and to lower risk population groups, should continue to include permanent reduction of dietary saturated fat and partial replacement by unsaturates. PMID- 10796867 TI - Chemotherapy for non-small cell lung cancer. AB - BACKGROUND: The role of chemotherapy in the treatment of patients with non-small cell lung cancer was not clear. A systematic review and quantitative meta analysis was therefore undertaken to evaluate the available evidence from all relevant randomised trials. OBJECTIVES: To evaluate the effect of cytotoxic chemotherapy on survival in patients with non-small cell lung cancer. To investigate whether or not pre-defined patient sub-groups benefit more or less from chemotherapy. SEARCH STRATEGY: MEDLINE and CANCERLIT searches were supplemented by information from trial registers and by hand searching relevant meeting proceedings and by discussion with relevant trialists and organisations. SELECTION CRITERIA: Trials comparing primary treatments of surgery, surgery + radiotherapy, radical radiotherapy or supportive care versus the same primary treatment, plus chemotherapy were eligible for inclusion provided that they randomised non-small cell lung cancer patients using a method which precluded prior knowledge of treatment assignment. DATA COLLECTION AND ANALYSIS: A quantitative meta-analysis using updated information from individual patients from all available randomised trials was carried out. Data from all patients randomised in all eligible trials were sought directly from those responsible. Updated information on survival, and date of last follow up were obtained, as were details of treatment allocated, date of randomisation, age, sex, histological cell type, stage and performance status. To avoid potential bias, information was requested for all randomised patients including those who had been excluded from the investigators' original analyses. All analyses were done on intention to treat on the endpoint of survival. For trials using cisplatin based regimens, subgroup analyses by age, sex, histological cell type, tumour stage and performance status were also done. MAIN RESULTS: Data from 52 trials and 9387 patients were included. The results for modern regimens containing cisplatin favoured chemotherapy in all comparisons and reached conventional levels of significance when used with radical radiotherapy and with supportive care. Trials comparing surgery with surgery plus chemotherapy gave a hazard ratio of 0.87 (13% reduction in the risk of death, equivalent to an absolute benefit of 5% at 5 years). Trials comparing radical radiotherapy with radical radiotherapy plus chemotherapy gave a hazard ratio 0.87 (13% reduction in the risk of death equivalent to an absolute benefit of 4% at 2 years), and trials comparing supportive care with supportive care plus chemotherapy gave a hazard ratio of 0.73 (27% reduction in the risk of death equivalent to a 10% improvement in survival at one year). The essential drugs needed to achieve these effects were not identified. No difference in the size of effect was seen in any subgroup of patients. In all but the radical radiotherapy setting, older trials using long term alkylating agents tended to show a detrimental effect of chemotherapy. This effect reached conventional significance in the adjuvant surgical comparison. REVIEWER'S CONCLUSIONS: At the outset of this meta-analysis there was considerable pessimism about the role of chemotherapy in the treatment of non small cell lung cancer. These results offer hope of progress and suggest that chemotherapy may have a role in treating this disease. PMID- 10796868 TI - Postoperative radiotherapy for non-small cell lung cancer. AB - BACKGROUND: The role of postoperative radiotherapy in the treatment of patients with completely resected non-small cell lung cancer was not clear. A systematic review and quantitative meta-analysis was therfore undertaken to evaluate the available evidence from randomised trials. OBJECTIVES: To evaluate the effect of post-operative radiotherapy (PORT) on survival and recurrence in patients with completely resected non-small cell lung cancer. To investigate whether or not pre defined patient sub-groups benefit more or less from PORT. SEARCH STRATEGY: MEDLINE and CANCERLIT searches were supplemented by information from trial registers and by hand searching relevant meeting proceedings and by discussion with relevant trialists and organisations. SELECTION CRITERIA: Both published and unpublished trials were eligible for inclusion provided the patients had undergone a complete resection; had been randomised between radiotherapy and no immediate further treatment; that the method of randomisation precluded prior knowledge of the treatment to be assigned; and that recruitment was between 1965 and 1995. DATA COLLECTION AND ANALYSIS: A quantitative meta-analysis using updated information from individual patients from all available randomised trials was carried out. Data from all patients randomised in all eligible trials were sought directly from those responsible. Updated information on survival, recurrence and date of last follow up were obtained, as were details of treatment allocated, date of randomisation, age, sex, histological cell type, stage and performance status. To avoid potential bias, information was requested for all randomised patients including those who had been excluded from the investigators' original analyses. All analyses were done on intention to treat on the endpoints of survival and recurrence-free survival. Subgroup analyses by age, sex, histological cell type, tumour stage and performance status were also done. MAIN RESULTS: 2128 patients from 9 trials were included (median follow-up of 3.9 years). The results show a significant adverse effect of PORT on survival with a hazard ratio of 1.21 or 21% relative increase in the risk of death. This is equivalent to an absolute detriment of 7% at 2 years (95% confidence interval 3 to 11%) reducing overall survival from 55% to 48%. Exploratory subgroup analyses suggested that this detrimental effect was most pronounced for patients with stage I/II, N0-N1 disease, whereas for stage III, N2 patients there was no clear evidence of an adverse effect. REVIEWER'S CONCLUSIONS: PORT is detrimental to patients with early stage completely resected non-small cell lung cancer and should not be used in the routine treatment of such patients. The role of PORT in the treatment of N2 tumours is not clear and may justify further research. PMID- 10796869 TI - Protein restriction for diabetic renal disease. AB - OBJECTIVES: To determine whether protein restriction slows or prevents progression of diabetic nephropathy towards renal failure. SEARCH STRATEGY: Computerised databases MEDLINE (1976-1996) and EMBASE (1974-1996) were searched using keywords diabetes mellitus, diabetic nephropathy, dietary proteins, diet, protein restricted and uremia. Recent issues of selected journals (Diabetic Medicine, Diabetologia, Diabetes Care, Kidney International, Nephrology Dialysis and Transplantation) were handsearched for papers not yet in the computerised databases. Reference lists of papers were also checked. SELECTION CRITERIA: This review was not limited to randomised controlled trials. All trials involving people with insulin-dependent diabetes following a lower protein diet for at least 4 months were considered since the straight line nature of progression as reflected by GFR means that patients can act as their own controls in a before and after comparison. DATA COLLECTION AND ANALYSIS: Data were extracted for length of follow up, level of protein restriction, renal function and dietary compliance. No studies of the impact of protein restriction on outcomes such as the need for dialysis or transplantation were found. The trials reported only the effect on short-term indicators such as creatinine clearance. MAIN RESULTS: Overall a protein restricted diet (0.3-0. 8g/kg) does appear to slow the progression of diabetic nephropathy towards renal failure. REVIEWER'S CONCLUSIONS: The results show that reducing protein intake appears to slow progression to renal failure, but some questions remain unanswered. The first is what level of protein restriction we should be used? The trials aimed for a daily intake of between 0.3 to 0.8g/kg of protein. The second concerns compliance in routine care - what level would be acceptable to patients? The third concerns long term outcomes -the present trials use proxy indicators such as creatinine clearance rather than outcomes such as time to dialysis or prevention of ESRF. All trials were carried out in subjects with insulin-dependent diabetes. It remains to be seen if a lower protein intake would slow the progression of nephropathy affecting the non-insulin dependent diabetic population. PMID- 10796870 TI - Aldose reductase inhibitors for the prevention and treatment of diabetic peripheral neuropathy. AB - OBJECTIVES: To assess the efficacy of aldose reductase inhibitors in the prevention, reversal or delay in the progression of diabetic peripheral neuropathy. SEARCH STRATEGY: The Cochrane Diabetes Group's database was searched and the citation lists of identified trials and previous reviews checked. Investigators identified as active in the field were approached for overlooked studies. SELECTION CRITERIA: Randomised controlled trials of aldose reductase inhibitors versus placebo, no treatment or other treatment in diabetic patients with or without clinical neuropathy. DATA COLLECTION AND ANALYSIS: Nerve conduction velocity was the only end point measured in all trials. Treatment effect was evaluated in terms of nerve conduction velocity mean difference in median and peroneal motor and median and sural sensory nerves. MAIN RESULTS: 19 trials, testing 4 different aldose reductase inhibitors for between 4 to 208 weeks duration (median 24 weeks), met the inclusion criteria for the meta analysis. A small but statistically significant reduction in decline of median and peroneal motor nerve conduction velocities was present in the treated group when compared to the control group (weighted mean 0.66 m/s 95% CI 0.18-1.14 m/s and 0.53 m/s 95% CI 0.02-1.04m/s respectively). No clear benefit of aldose reductase inhibitor treatment was observed in either of the sensory nerves. REVIEWER'S CONCLUSIONS: Although aldose reductase inhibitor treatment has been demonstrated to diminsh the reduction in motor nerve conduction velocity, the clinical relevance of such a change in this outcome measure is uncertain. There was no effect in terms of this outcome measure in the smaller sensory fibres, degeneration of which is primarily responsible for the most common neuropathic syndrome associated with diabetes, that of severe pain and loss of sensation in the extremity leading in some cases to ulceration and eventual amputation. PMID- 10796871 TI - Angiotensin converting enzyme inhibitors in normotensive diabetic patients with microalbuminuria. AB - OBJECTIVES: To examine whether the progression of early diabetic renal disease to end-stage renal failure may be slowed by the use of angiotensin converting enzyme inhibitors for reasons other than their antihypertensive properties, so that they have value in the treatment of normotensive diabetics with microalbuminuria. SEARCH STRATEGY: MEDLINE was searched for English language reviews and randomised controlled trials. Personal reference lists, and reference lists of retrieved studies were also used. SELECTION CRITERIA: Randomised controlled trials with separate identifiable results for initially normotensive diabetic patients, who received angiotensin converting enzyme inhibitors for at least one year and were compared with controls. DATA COLLECTION AND ANALYSIS: Meta-analyses were performed on the results of 11 randomised controlled trials with a variety of patient inclusion and exclusion criteria. MAIN RESULTS: Albumin excretion rate fell for patients on angiotensin converting enzyme inhibition in 10 of the 11 studies but did so for only two of the 11 groups on placebo. Treatment provided a significant reduction in albumin excretion rate in both insulin dependent diabetes mellitus and non insulin dependent diabetes mellitus. Treatment with either captopril, enalapril or lisinopril reduced albumin excretion rate in comparison with control patients. A greater lowering of blood pressure was experienced by initially normotensive patients in the angiotensin converting enzyme inhibitor than in the placebo group. Pooled end-of-study mean blood pressures for the treated group were significantly lower than for the untreated group, although this was not so for all individual studies. The apparent (treatment - placebo) 'effect' on systolic and diastolic pressures had a larger standard error for longer lasting studies. Average haemoglobin fell a little in the treated patients and rose in the controls but neither this nor the difference in changes in glomerular filtration rate reached statistical significance. REVIEWER'S CONCLUSIONS: Inhibition of angiotensin converting enzyme can arrest or reduce the albumin excretion rate in microalbuminuric normotensive diabetics, as well as reduce or prevent an increase in blood pressure. But, given the drop in blood pressure in patients on angiotensin converting enzyme inhibitors, it is not possible to be certain that the reduction of albumin excretion rate is due to a separate renal effect. A direct link with postponement of end-stage renal failure has not been demonstrated. There appear to be no substantial side effects. PMID- 10796872 TI - Antihypertensive therapy for preventing cardiovascular complications in people with diabetes mellitus. AB - OBJECTIVES: To assess the effect of intervention, both pharmacological and non parmacological, to reduce blood pressure in people with diabetes mellitus on all cause mortality, specific causes of death, including cardiovascular disease, stroke, ischaemic heart disease and renal disease, morbidity associated with macro- and microvascular complications of diabetes mellitus and also side effects of the interventions and their influence on quality of life and well being. SEARCH STRATEGY: The search strategy employed was to searching electronic databases such as EMBASE and MEDLINE for all trials of anti-hypertensive treatment in diabetes mellitus. As well as searching specialist journals in the fields of cardiovascular disease, stroke, hypertension and renal diease. SELECTION CRITERIA: All trials were considered independently and then discussed by 2 reviewers to determine there eligibility for inclusion in the review. Their methodological quality was also assessed from details of the randomisation methods, blinding and whether the intention-to-treat method of analysis was used. Trials included in the review were all randomised contolled trials of the treatment for anti-hypertensive therapy for the specified endpoints which included subjects with diabetes mellitus. DATA COLLECTION AND ANALYSIS: Data was sought on the number of patients with diabetes with each outcome measure by allocated treatment group, either from previous publications or, if this was not possible, the raw data was obtained and analysed using the intention-to-treat method. If these data were not available the results from the 'Per Protocol' analysis were used. To compare the treatment effect of the intervention with that of placebo on all cause mortality and cardiaovascular mortality and morbidity, odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for each trial and a meta analysis performed using Peto's ORs as the summary measure. MAIN RESULTS: The initial search yielded 760 references, from which 23 appropriate trials were identified (3 for primary prevention and 20 for secondary prevention), and 15 of these trials had data available for analysis. For the primary prevention trials the summary ORs (95% CIs) for all cause mortality and CVD were 0.85 (0.62,1.17) and 0.64 (0.50,0.82) respectively. Of the seven trials for long-term secondary prevention (i.e. follow-up greater than one year), the summary OR (95% CI) for all cause mortality was 0.82 (0.69,0.99). Data on CVD mortality and morbidity was only available for 2 of these trials and the summary OR (95% CI) was 0.82 (0.60,1.13). There were five trials for short term secondary prevention trials (i.e. follow-up of less than 1 year) with data available for analysis. The summary ORs (95% CIs) for all cause mortality and CVD were 0.64 (0.50,0.83) and 0.68 (0.43,1.05) respectively. REVIEWER'S CONCLUSIONS: Primary intervention trials indicated a treatment benefit for CVD, but not for total mortality in people with diabetes. For both short- and long-term secondary prevention, the present meta-analysis indicated a benefit for total mortality in diabetic subjects. However lack of information on CVD outcomes probably reduced the power of the meta-analysis to detect any corresponding benefit for this end point. This, along with the fact that all published data of randomised control trials of anti-hypertensive therapy in diabetes for all cause mortailty and CVD outcomes are taken from the hypertension trials not specific to diabetes, underlines the need for further high quality trials examining the effects of blood pressure lowering interventions in people with diabetes. PMID- 10796873 TI - Adjuvant chemotherapy for localised resectable soft tissue sarcoma in adults. Sarcoma Meta-analysis Collaboration (SMAC). AB - BACKGROUND: Individually, randomised trilas have not shown conclusively whether adjuvant chemotherapy benefits adult patients with localised resectable soft tissue sarcoma. OBJECTIVES: Adjuvant chemotherapy aims to lessen the recurrence of cancer after surgery with or without radiotherapy. The objective of this review was to assess the effects of adjuvant chemotherapy in adults with resectable soft tissue sarcoma after such local treatment. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, UKCCCR Register of Cancer Trials, Physicians Data Query, EMBASE, MEDLINE and CancerLit. SELECTION CRITERIA: Randomised trials of adjuvant chemotherapy after local treatment in adults with localised resectable soft tissue sarcoma were included. Only trials in which accrual was completed by December 1992 were included. DATA COLLECTION AND ANALYSIS: Individual patient data were obtained. Accuracy of data and quality of randomisation and follow-up of trials was assessed. MAIN RESULTS: Fourteen trials of doxorubicin-based adjuvant chemotherapy involving 1568 patients were included. Median follow-up was 9.4 years. For local recurrence-free interval the hazard ratio with chemotherapy was 0.73 (95% Confidence Interval 0.56-0.94). For distant recurrence-free interval it was 0. 70 (95% CI 0.57-0.85). For overall recurrence free survival it was 0. 75 (95% CI 0.64-0.87). These correspond to significant absolute benefits of 6-10% at 10 years. For overall survival the hazard ratio of 0.89 (95% CI 0.76-1.03) was not significant but potentially represents an absolute benefit of 4% (95% CI -1 to 9) at 10 years. There was no consistent evidence of a difference in effect according to age, sex, stage, site, grade, histology, extent of resection, tumour size or exposure to radiotherapy. However, the strongest evidence of a beneficial effect on survival was shown in patients with sarcoma of the extremities. REVIEWER'S CONCLUSIONS: Doxorubicin-based adjuvant chemotherapy appears to significantly improve time to local and distant recurrence and overall recurrence-free survival in adults with localised resectable soft tissue sarcoma. There is some evidence of a trend towards improved overall survival. PMID- 10796875 TI - Analysis and classification of 304 mutant alleles in patients with type 1 and type 3 Gaucher disease. AB - Gaucher disease results from the inherited deficiency of the enzyme glucocerebrosidase (EC 3.2.1.45). Although >100 mutations in the gene for human glucocerebrosidase have been described, most genotype-phenotype studies have focused upon screening for a few common mutations. In this study, we used several approaches-including direct sequencing, Southern blotting, long-template PCR, restriction digestions, and the amplification refraction mutation system (ARMS) to genotype 128 patients with type 1 Gaucher disease (64 of Ashkenazi Jewish ancestry and 64 of non-Jewish extraction) and 24 patients with type 3 Gaucher disease. More than 97% of the mutant alleles were identified. Fourteen novel mutations (A90T, N117D, T134I, Y135X, R170C, W184R, A190T, Y304X, A341T, D399Y, c.153-154insTACAGC, c.203-204insC, c.222-224delTAC, and c.1122-1123insTG) and many rare mutations were detected. Recombinant alleles were found in 19% of the patients. Although 93% of the mutant alleles in our Ashkenazi Jewish type 1 patients were N370S, c.84-85insG, IVS2+1G-->A or L444P, these four mutations accounted for only 49% of mutant alleles in the non-Jewish type 1 patients. Genotype-phenotype correlations were attempted. Homozygosity or heterozygosity for N370S resulted in type 1 Gaucher disease, whereas homozygosity for L444P was associated with type 3. Genotype L444P/recombinant allele resulted in type 2 Gaucher disease, and homozygosity for a recombinant allele was associated with perinatal lethal disease. The phenotypic consequences of other mutations, particularly R463C, were more inconsistent. Our results demonstrate a high rate of mutation detection, a large number of novel and rare mutations, and an accurate assessment of the prevalence of recombinant alleles. Although some genotype-phenotype correlations do exist, other genetic and environmental factors must also contribute to the phenotypes encountered, and we caution against relying solely upon genotype for prognostic or therapeutic judgements. PMID- 10796874 TI - Parametric and nonparametric multipoint linkage analysis with imprinting and two locus-trait models: application to mite sensitization. AB - We present two extensions to linkage analysis for genetically complex traits. The first extension allows investigators to perform parametric (LOD-score) analysis of traits caused by imprinted genes-that is, of traits showing a parent-of-origin effect. By specification of two heterozygote penetrance parameters, paternal and maternal origin of the mutation can be treated differently in terms of probability of expression of the trait. Therefore, a single-disease-locus imprinting model includes four penetrances instead of only three. In the second extension, parametric and nonparametric linkage analysis with two trait loci is formulated for a multimarker setting, optionally taking imprinting into account. We have implemented both methods into the program GENEHUNTER. The new tools, GENEHUNTER-IMPRINTING and GENEHUNTER-TWOLOCUS, were applied to human family data for sensitization to mite allergens. The data set comprises pedigrees from England, Germany, Italy, and Portugal. With single-disease-locus-imprinting MOD score analysis, we find several regions that show at least suggestive evidence for linkage. Most prominently, a maximum LOD score of 4.76 is obtained near D8S511, for the English population, when a model that implies complete maternal imprinting is used. Parametric two-trait-locus analysis yields a maximum LOD score of 6.09 for the German population, occurring exactly at D4S430 and D18S452. The heterogeneity model specified for analysis alludes to complete maternal imprinting at both disease loci. Altogether, our results suggest that the two novel formulations of linkage analysis provide valuable tools for genetic mapping of multifactorial traits. PMID- 10796876 TI - Null alleles of the COL5A1 gene of type V collagen are a cause of the classical forms of Ehlers-Danlos syndrome (types I and II). AB - Ehlers-Danlos syndrome (EDS) types I and II, which comprise the classical variety, are well characterized from the clinical perspective, but it has been difficult to identify the molecular basis of the disorder in the majority of affected individuals. Several explanations for this failure to detect mutations have been proposed, including genetic heterogeneity, failure of allele expression, and technical difficulties. Genetic heterogeneity has been confirmed as an explanation for such failure, since causative mutations have been identified in the COL5A1, COL5A2, and tenascin X genes and since they have been inferred in the COL1A2 gene. Nonetheless, in the majority of families with autosomal dominant inheritance of EDS, there appears to be linkage to loci that contain the COL5A1 or COL5A2 genes. To determine whether allele-product instability could explain failure to identify some mutations, we analyzed polymorphic variants in the COL5A1 gene in 16 individuals, and we examined mRNA for the expression of both alleles and for alterations in splicing. We found a splice-site mutation in a single individual, and we determined that, in six individuals, the mRNA from one COL5A1 allele either was not expressed or was very unstable. We identified small insertions or deletions in five of these cell strains, but we could not identify the mutation in the sixth individual. Thus, although as many as one-half of the mutations that give rise to EDS types I and II are likely to lie in the COL5A1 gene, a significant portion of them result in very low levels of mRNA from the mutant allele, as a consequence of nonsense mediated mRNA decay. PMID- 10796877 TI - Clinical relevance of hepatitis B viral mutations. PMID- 10796879 TI - An exploratory population-based case-control study of primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a disease of unknown etiology, with unexplained geographical variation. Various exposures have been suggested as triggers for disease development-possibly in susceptible individuals, but the evidence was not always well founded. We therefore conducted a population-based case-control study in Northeast England to investigate these and other exposures. All cases incident during 1993 to 1995 in a defined area of Northeast England were identified, and age- and sex-matched population controls were identified from primary care population registers. Cases and controls were sent postal self completion questionnaires covering medical history and lifestyle. Information was received from 100 cases and 223 controls. The familial tendency of PBC was found to be less marked than has been claimed: Only weak associations were found with other autoimmune diseases. Among factors considered previously, no significant associations were found with surgical procedures, events in pregnancy, past infections, vaccinations, and medications. No significant associations were found for previously unconsidered lifestyle factors (drinking alcohol, previous pets, or stressful events), but there was an unexpected association with past smoking (ever smoked: 76% in cases vs. 57% in controls, odds ratio 2.4; smoked for 20 years or more: 64% vs. 35%, odds ratio 3.5). There were also unexpected significant associations with psoriasis (13% in cases vs. 3% in controls, odds ratio 4.6) and eczema (3% in cases vs. 11% in controls, odds ratio 0. 13). These findings merit further investigation. PMID- 10796878 TI - ATP depletion in rat cholangiocytes leads to marked internalization of membrane proteins. AB - Intrahepatic bile ducts (BD) are a critical target of injury in the postischemic liver. Decreased vascular perfusion causes characteristic changes in the morphology of the ductular epithelia including a loss of secondary membrane structures and a decrease in plasma membrane surface area. Using adenosine triphosphate (ATP) depletion of cultured normal rat cholangiocytes (NRC) to model ischemic ducts, the present studies examined the fate of apical membrane proteins to determine whether membrane recycling might contribute to rapid functional recovery. Apical proteins, including gamma-glutamyl transpeptidase (GGT), Na(+) glucose cotransporter (SGLT1), and apically biotinylated proteins, were not shed into the luminal space during ATP depletion. Instead, labeling of surface proteins after ATP depletion showed a significant decrease in GGT and SGLT1, consistent with membrane internalization. Similarly, z-axis confocal microscopy of biotinylated apical proteins also showed protein internalization. During ATP recovery, SGLT1 transport activity remained profoundly depressed even after 24 hours of recovery, indicating that the function of the internalized apical proteins is not rapidly recovered. These studies suggest that the membrane internalization in ATP-depleted cholangiocytes is a unidirectional process that contributes to prolonged functional deficits after restoration of normal cellular ATP levels. This sustained decrease in transport capacity may contribute to the development of ductular injury in postischemic livers. PMID- 10796880 TI - Predictive value of the variceal pressure response to continued pharmacological therapy in patients with cirrhosis and portal hypertension. AB - Noninvasive measurements of variceal pressure adequately reflect the hemodynamic effects of propranolol on portal hypertension. However, the prognostic value of variceal pressure responses during continued propranolol therapy has not been evaluated, and it is unclear whether this may substitute invasive measurements of portal pressure response. Fifty-five portal hypertensive patients with cirrhosis were studied before and at 4 months of continued propranolol therapy. Variceal pressure was measured using an endoscopic pressure gauge. Portal pressure was evaluated as the hepatic venous pressure gradient (HVPG). Over a 28 +/- 11 month follow-up, 16 patients experienced variceal bleeding. Baseline characteristics were similar in bleeders and nonbleeders. At 4 months, reduction in variceal pressure was less marked in bleeders than in nonbleeders (5% +/- 20% vs. -15% +/- 24%; P =.03). A fall in variceal pressure 20% or greater of baseline was an independent predictor of absence of variceal bleeding; which occurred in 5% of patients with a 20% or greater fall in variceal pressure versus 42% of patients with less than a 20% reduction (P =.004). The HVPG response had similar independent prognostic value (decrease > or =20%: 6% bleeding; decrease <20%: 45% bleeding; P =.004) but identified different patients. Achieving a 20% decrease in either variceal pressure or HVPG was highly sensitive (85%) and specific (93%) identifying patients not bleeding on follow-up. Endoscopic measurements of variceal pressure response to continued pharmacotherapy provide useful prognostic information on the risk of variceal bleeding. As with HVPG response, a fall in variceal pressure of 20% or greater is associated with a very low risk of variceal bleeding. The combination of both parameters allows almost optimal prognostication. PMID- 10796881 TI - p27(Kip1) expression in normal epithelia, precancerous lesions, and carcinomas of the gallbladder: association with cancer progression and prognosis. AB - p27(Kip1) is a cyclin-dependent kinase inhibitor that negatively regulates cell proliferation. This study was designed to evaluate the roles of p27(Kip1) in gallbladder carcinogenesis and the prognostic value of p27(Kip1) in patients with gallbladder carcinoma. p27(Kip1) expression was examined immunohistochemically in surgically resected specimens of 8 normal epithelia, 8 adenomyomatosis lesions, 6 precancerous adenomas, and 37 carcinomas of the gallbladder. Decreased p27(Kip1) expression (<50% nuclear staining) was observed in 16 of the 37 (43%) gallbladder carcinomas, but not in any specimen of normal epithelium, adenomyomatosis, or adenoma. The fact that all of the adenomas showed normal p27(Kip1) expression suggests that decreased p27(Kip1) expression is probably not an early event in gallbladder carcinogenesis. Decreased p27(Kip1) expression was significantly associated with less marked tumor cell differentiation (P =.017), lymphatic invasion (P =.046), lymph node metastasis (P =.007), and advanced TNM stage (stage IV vs. stage I, P =.026; stage IV vs. stage II, P =.005). This suggests that down-regulation of p27(Kip1) expression is a late event in gallbladder carcinogenesis, possibly promoting tumor progression and metastasis. Kaplan-Meier curves showed that decreased p27(Kip1) expression was significantly associated with shorter overall survival (P =.001) in patients with gallbladder carcinomas who had undergone radical surgery. Cox's proportional hazards model revealed decreased p27(Kip1) expression to be an independent predictor for death (P =.034; risk ratio, 3.9; 95% confidence interval, 1.1-13.7). In conclusion, decreased p27(Kip1) expression significantly correlates with tumor progression and predicts poor prognosis in gallbladder carcinomas. PMID- 10796882 TI - Comprehensive allelotype study of hepatocellular carcinoma: potential differences in pathways to hepatocellular carcinoma between hepatitis B virus-positive and negative tumors. AB - To examine the role of the loss of heterozygosity (LOH) in hepatitis-related carcinogenesis, we performed a genome-wide scan of LOH in 44 tumors of hepatocellular carcinoma (HCC) using 216 microsatellite markers throughout all human chromosomes. A high frequency of LOH (>30% of informative cases) was observed at 33 loci on chromosome arms 4q, 6q, 8p, 8q, 9p, 9q, 13q, 16p, 16q, 17p, and 19p. LOH on 19p has not yet been reported, and that appears to be a new candidate in the search for tumor suppressor genes. High rates of LOH are correlated with hepatitis B virus (HBV) positivity, poorly differentiated tumors, vascular invasion, and intrahepatic metastasis (P <.0001). LOH on 13q and 16q occurred more frequently in HBV(+) patients (P <.0001), and LOH on 6q occurred more frequently in virus-negative patients (P <.001). The frequency of LOH on 4q and 13q was significantly lower in well-differentiated tumors than in moderately and poorly differentiated tumors (P <.01). In contrast, LOH on 6q was frequently detected in well-differentiated tumors compared with other histological subclasses (P <.001). Our results suggest that LOH on 6q may play an important role in the early stage of hepatocarcinogenesis in virus-negative patients, but different mechanisms might underlie the initial step to carcinogenesis in HBV(+) patients. LOH on 13q and 16q may play an essential role in the progression of HBV(+) tumors. Further studies of fine deletion mapping on chromosomes 13q and 16q are required to define the genomic segments on which putative tumor suppressor genes responsible for HBV(+) tumors exist. PMID- 10796883 TI - Survivin promotes cell proliferation in human hepatocellular carcinoma. AB - Survivin is a recently described inhibitor of apoptosis. Because suppression of apoptosis is important for carcinogenesis and tumor growth, we investigated the expression and function of survivin in human hepatocellular carcinomas (HCCs). We have shown that 4 HCC cell lines and 7 out of 8 human HCC tissues expressed survivin messenger RNA (mRNA), whereas expression of survivin mRNA was not detected in normal liver and nontumor areas of these tissues using the reverse transcription polymerase chain reaction. Survivin was detected primarily in the nucleus by immunofluorescence staining of HCC cells. In addition, 14 of 20 (70%) HCC tissues showed positive nuclear staining for survivin, whereas nontumor tissues showed little detectable staining by immunohistochemistry. Survivin expression strongly correlated with the proliferation index but not significantly with the apoptosis index in HCC tissues. Therefore, we performed cell cycle analysis after survivin transfection and showed that overexpression of survivin resulted in a decrease in the G(0)/G(1) phase and an increase in the S phase in all 4 HCC cell lines. Furthermore, we have found that survivin interacted with cyclin-dependent kinase 4 (Cdk4) and overexpression of survivin released p21(WAF1/Cip1) (p21) from Cdk4. From these results, we conclude that survivin promotes cell proliferation by interacting with Cdk4 and releasing p21 from Cdk4. This may play an important role in carcinogenesis and progression of human HCCs. PMID- 10796884 TI - Tumor necrosis factor alpha down-regulates expression of the alpha1(I) collagen gene in rat hepatic stellate cells through a p20C/EBPbeta- and C/EBPdelta dependent mechanism. AB - Tumor necrosis factor alpha (TNF-alpha) is one of the key cytokines of the acute phase response and of many inflammatory processes. This cytokine has several antifibrogenic actions and down-regulates the expression of the type I collagen genes and induces the expression of metalloproteinases. Because TNF-alpha directly antagonizes some fibrogenic actions of transforming growth factor beta(1) (TGF-beta(1)), we considered it important to map the cis-acting regulatory element of the alpha1(I) collagen (col1a1) promoter involved in TNF alpha responsiveness in hepatic stellate cells (HSC), to investigate the transcription factors that bind to it, and to establish possible mechanisms by which TNF-alpha down-regulates its expression. In this article, we show the presence of a functional TNF-alpha-responsive element (TaRE) in the -378 to -345 region of the col1a1 promoter. This element colocalizes with a previously reported TGF-beta(1)-responsive element. We further demonstrate that TNF-alpha induces nuclear translocation and binding of transcriptional complexes containing p20C/EBPbeta, p35C/EBPbeta, and C/EBPdelta to this sequence of the promoter. Transient overexpression of C/EBPdelta or p20C/EBPbeta, the natural dominant negative form of C/EBPbeta in HSC, down-regulated activity of a CAT reporter vector driven by -412 to +110 of the col1a1 promoter. Taken together, these data suggest that the -378 to -340 region of the col1a1 promoter is the site of convergence of different stimuli that ultimately modulate col1a1 gene transcription. PMID- 10796886 TI - Production and release of endothelin-1 from the gut and spleen in portal hypertension due to cirrhosis. AB - This study was aimed to evaluate the source of endothelin-1 (ET-1) in cirrhotic patients. ET-1 is implicated in the pathogenesis of portal hypertension. However, the mechanism and source for increased plasma ET-1 in cirrhotic patients are still obscure. Plasma ET-1 levels in systemic (SV), superior mesenteric (SMV), and splenic venous (SPV) blood were measured in 23 patients with cirrhosis and 8 controls with normal liver. Fourteen removed spleens were immunohistochemically studied for ET-1, CD34, CD68, and CD20. In situ hybridization was done to localize ET-1 messenger RNA (mRNA). In cirrhosis, ET-1 levels in both SMV and SPV were higher than in SV. ET-1 in SV and SPV were significantly higher in cirrhotic patients than in control patients. Three groups of cells in the spleen expressed both protein and mRNA of ET-1: endothelial cells in the sinus, which were also stained for CD34; cells in the germinal center; and cells in the marginal zone of lymphoid sheaths and follicles, which were also stained for CD20 but not for CD34 and CD68. The ET-1 concentration released from the spleen was in parallel with the grade of ET-1 expression in the spleen. The spleen is one of the major sites of ET-1 release in cirrhotic patients. Endothelial cells of the splenic sinus and possibly B lymphocytes in the germinal center and marginal zone of lymphoid sheaths and follicles seem to be the sites of ET-1 production in the spleen. PMID- 10796885 TI - Modulation of transforming growth factor beta response and signaling during transdifferentiation of rat hepatic stellate cells to myofibroblasts. AB - Activation of hepatic stellate cells (HSCs) is the key step in liver fibrogenesis. Increased transforming growth factor beta (TGF-beta) expression and extracellular matrix production in patients with hepatic fibrosis and experimental models of liver fibrogenesis support implication of TGF-beta in the pathogenesis of this disease. However, a causative role for TGF-beta during transdifferentiation of HSCs has not been delineated in molecular detail. Using a rat cell culture model of HSC transdifferentiation, we analyzed TGF-beta signal transduction and identified changes between stellate cells and their transdifferentiated phenotype. Fully transdifferentiated myofibroblasts, opposed to HSCs, were not inhibited in proliferation activity on treatment with TGF beta1. Furthermore, stimulation of alpha2 (I) collagen and Smad7 messenger RNA (mRNA) expression by TGF-beta1 was achieved in stellate cells but not in myofibroblasts. Northern and Western blot analyses indicated significant expression of TGF-beta receptors I and II in both cell types. In contrast, [(125)I]-TGF-beta1 receptor affinity labeling displayed strongly reduced types I, II, and III receptor presentation at the cell surface of myofibroblasts. Moreover, myofibroblasts did not display DNA-binding SMAD proteins in electrophoretic mobility shift assays with a CAGA box. These data indicate that stellate cells are responsive to TGF-beta1 treatment and transduce a signal that may play an important role in liver fibrogenesis. Myofibroblasts display decreased availability of surface receptors for TGF-beta, which could be based on autocrine stimulation. However, lack of activated SMAD complexes with DNA-binding activity and absence of alpha2 (I) collagen transcription inhibition by latency associated peptide (LAP)/anti-TGF-beta antibody raise the possibility of TGF-beta signaling independent receptor down-regulation in myofibroblasts. PMID- 10796887 TI - Glutathione, lactobionate, and histidine: cryptic inhibitors of matrix metalloproteinases contained in University of Wisconsin and histidine/tryptophan/ketoglutarate liver preservation solutions. AB - UW solution and HTK solution are both used for cold preservation of liver allografts. Although they are about equally effective, their compositions are very different, and they were formulated using different rationales. The authors recently showed an important role for MMPs in liver preservation injury and consequently postulated that these preservation solutions contain cryptic inhibitors of MMP activity. To determine this possibility, the ability of these solutions to inhibit MMP activity was studied. The source of MMP2 and MMP9 was human liver effluents obtained at the time of liver transplantation or commercially available human recombinant MMP2 and MMP9. MMP2 and MMP9 showed gelatinolytic activity at 37 degrees C and also at 4 degrees C, although activity at 4 degrees C was reduced. Activity was inhibited by University of Wisconsin (UW) and Histidine/Tryptophan/ Ketoglutarate (HTK) solutions. Examination of individual ingredients disclosed that reduced glutathione (GSH) and lactobionate in UW solution and histidine in HTK solution were the cryptic inhibitors. HTK solution was a more effective inhibitor than UW solution. GSH inhibited the activity of both enzymes, but was a much more effective inhibitor of MMP9 than MMP2. Oxidized glutathione(GSSG) was a much less effective inhibitor of the enzymes. The inhibitor constants (K(i)) of GSH for MMP2 and MMP9 were 34 micromol/L and 3 micromol/L, respectively. The authors conclude that MMP inhibition is a cryptic property of both commonly used liver preservation solutions and contributes importantly to their action. Furthermore, GSH appears to be an effective inhibitor of gelatinases at concentrations at which it is normally present in extracellular fluid. PMID- 10796888 TI - Molecular mechanisms of tumor necrosis factor alpha-stimulated leukocyte recruitment into the murine hepatic circulation. AB - To date, much of the adhesion work in the liver has been restricted to sinusoids and postsinusoidal venules. However, selectins have been localized on the portal (presinusoidal) venules and these vessels have been shown to be important in metastasis of tumors. The purpose of this study was to characterize the leukocyte endothelial interactions within the 3 compartments of the hepatic microvasculature under baseline conditions and in response to tumor necrosis factor alpha (TNF-alpha). Mice deficient in P-selectin or both E- and P-selectin were compared with wild-type (C57Bl/6, wild type) mice. Animals were injected with murine TNF-alpha (15 microg/kg intraperitoneally [IP]) and the liver was examined by fluorescence intravital microscopy 4 hours later. Under baseline conditions, leukocyte flux in the portal venules was 1.42 +/- 0.42 cells/min. Leukocyte flux in the portal venules of wild-type mice increased 8-fold in response to 4 hours of TNF-alpha stimulation. This was reduced by 50% in the P selectin-deficient mice but was not reduced further by either the addition of an E-selectin antibody (9A9, 100 microg intravenously [IV]) to these mice or in mice deficient in both E- and P-selectin. In P-selectin-deficient mice, the addition of an antibody against alpha(4)-integrin (R1-2, 75 microg IP) reduced rolling to baseline. But in the E- and P-double-selectin-deficient mice the addition of an antibody against L-selectin (Mel 14, 3 microg/kg IV) had no effect on TNF-alpha induced recruitment. Similar responses were seen in the central venules, however, in the sinusoids the increased number of stationary leukocytes seen in response to 4 hours of TNF-alpha stimulation in the wild-type mice was not reduced in P selectin-deficient mice with or without the alpha(4)-integrin antibody. These data suggest that leukocytes can use alpha(4)-integrin independent of the selectins in the venules. Within the sinusoids, however, inhibition of E selectin, P-selectin, and alpha(4)-integrin was insufficient to reduce leukocyte recruitment. PMID- 10796889 TI - Induction by interleukin 6 of G(s)-coupled prostaglandin E(2) receptors in rat hepatocytes mediating a prostaglandin E(2)-dependent inhibition of the hepatocyte's acute phase response. AB - Prostanoids, that are released from nonparenchymal liver cells in response to proinflammatory stimuli, are involved in the regulation of hepatic functions during inflammation. They exert their effects on their target cells via heptahelical receptors in the plasma membrane. For the 5 prostanoids prostaglandin E(2) (PGE(2)), prostaglandin F(2alpha), prostaglandin D(2) (PGD(2)), prostacyclin, and thromboxane A(2) there exist 8 receptors that are coupled to different heterotrimeric G proteins. These receptors are expressed differentially in the 4 principal liver cell types, i.e., hepatocytes, Kupffer cells, sinusoidal endothelial cells, and hepatic stellate cells. It was intriguing, that the messenger RNA (mRNA) of none of the G(s)-coupled prostanoid receptors (DP-R, EP2-R, EP4-R, and IP-R) that can attenuate the inflammatory reaction were present in hepatocytes. The current study shows that the expression of the G(s)-coupled prostanoid receptors EP2-R, EP4-R, and DP-R, but not the IP R, was efficiently and rapidly up-regulated by treatment of hepatocytes in vitro or rats in vivo with the key acute phase cytokine interleukin 6 (IL-6). In IL-6 treated hepatocytes PGE(2) in turn attenuated the IL-6-induced alpha(2) macroglobulin formation via a cyclic adenosine monophosphate (cAMP)-dependent signal chain. The data indicate that an IL-6-mediated induction of the previously not expressed EP2-R and EP4-R on hepatocytes might establish a prostanoid mediated feedback inhibition loop for the attenuation of the acute phase response. PMID- 10796890 TI - Hypogonadism precedes liver feminization in chronic alcohol-fed male rats. AB - Men who chronically abuse alcohol may display a spectrum of endocrine abnormalities including hypogonadism and feminization, with elevated serum estradiol and low serum testosterone. We examined factors that may result in disruption of hepatic sex hormone homeostasis in alcohol-fed male rats and possible consequences of such changes. Rats were fed alcohol-containing or isocaloric diets for 30, 60, and 90 days. In alcohol-fed rats, serum testosterone levels and hepatic activity of 2 androgen-dependent estrogen metabolizing enzymes were reduced (P <.05) at all times, as was activity of androgen receptor. There was also a significant early and progressive decrease in testes/body ratio in alcohol-fed rats. Compared with this early decrease in testosterone-related parameters, there was a significant increase in serum estrogen levels (at 30 and 90 days, 132% and 168% of control values, respectively). An increase in serum ceruloplasmin, an estrogen-responsive liver protein, was apparent at 60 and 90 days, but not at 30 days of alcohol exposure, suggesting that hypogonadism precedes liver feminization. Hepatic estrogen receptor activity was decreased in alcohol-fed rats at 60 and 90 days, the latter despite elevated serum estrogen levels. Hepatic aromatase was slightly increased in alcohol-fed rats, an elevation probably not sufficient to account for observed increases in serum estrogen. Taken together, these data suggest that (1) alcohol induces profound reduction of serum testosterone, resulting in loss of androgen-regulated hepatic functions such as estrogen-metabolizing enzyme activity and activity of androgen receptors; and (2) such alcohol-induced hypogonadism precedes changes in hepatic sex hormone homeostasis and subsequent feminization. PMID- 10796891 TI - Ethanol potentiates tumor necrosis factor-alpha cytotoxicity in hepatoma cells and primary rat hepatocytes by promoting induction of the mitochondrial permeability transition. AB - In the present study, tumor necrosis factor-alpha (TNF-alpha) cytotoxicity is shown to be potentiated by ethanol exposure in vitro in the human hepatoma cell line, HepG2, and in rat primary hepatocytes. Exposure of HepG2 cells and primary hepatocytes for 48 hours to concentrations of ethanol ranging between 50 and 100 mmol/L significantly increased TNF-alpha cytotoxicity compared with cells treated with TNF-alpha alone. The cell killing was associated with, and dependent on, the development of the mitochondrial permeability transition (MPT). Two inhibitors of MPT pore opening, cyclosporin A and bongkrekic acid, prevented TNF-alpha cytotoxicity in the presence of ethanol. In addition to inhibiting cell death caused by TNF-alpha, blockade of MPT pore opening prevented mitochondrial depolarization, cytochrome c redistribution from the mitochondria to the cytosol, caspase 3 activation, and oligonucleosomal DNA fragmentation. Unlike the potentiation of TNF-alpha cytotoxicity by the translational inhibitor cycloheximide, ethanol promoted TNF-alpha-induced cell killing by a mechanism that was independent of caspase-8 activity. HepG2 cells overexpressing cytochrome P4502E1 were even more sensitized by ethanol to induction of the MPT by TNF-alpha and the resultant cytotoxicity than wild-type HepG2 cells. In addition, primary hepatocytes isolated from chronically ethanol-fed rats showed enhanced susceptibility to TNF-alpha cytotoxicity compared with their isocalorically matched controls. Again as with the HepG2 cells, inhibiting MPT pore opening prevented the cytotoxicity of TNF-alpha in the primary hepatocytes isolated from ethanol-fed animals. PMID- 10796892 TI - Liver cell apoptosis in chronic hepatitis C correlates with histological but not biochemical activity or serum HCV-RNA levels. AB - In hepatitis C virus (HCV) infection, mechanisms responsible for liver cell damage are still poorly understood and both necrosis and apoptosis may be operative. By using terminal deoxynucleotydil transferase-mediated d-UTP-biotin nick-end labeling (TUNEL) we have evaluated and quantified apoptosis in liver biopsy specimens from 61 patients with chronic hepatitis C. All patients had detectable apoptotic cells in the liver. Presence of increased apoptotic activity was confirmed in selected cases by electron microscopy and by DNA gel electrophoresis. The amount of liver cell apoptosis expressed as apoptotic index, ranged between 0.01% to 0.54% and showed a positive correlation with histological activity grading (P <.0005) and with the amount of infiltrating CD8-positive cells (P =. 01). Apoptosis did not correlate with transaminase levels or with HCV load and genotype. These results support the concept that immune-mediated apoptosis may play a role in the pathogenesis of chronic hepatitis C and indicate that this type of reaction may occur in the absence of significant alanine transaminase (ALT) elevation, thus explaining the lack of correlation between biochemical activity and liver histological damage. PMID- 10796893 TI - Population screening for hemochromatosis: a comparison of unbound iron-binding capacity, transferrin saturation, and C282Y genotyping in 5,211 voluntary blood donors. AB - Early diagnosis and treatment of hemochromatosis is essential to prevent organ damage. Screening strategies to detect early hemochromatosis include testing for iron overload and/or genetic testing. Voluntary blood donors numbering 5,211 were screened with unbound iron-binding capacity (UIBC), transferrin saturation (TS), and genetic testing for the C282Y mutation of the HFE gene. The study found 16 C282Y homozygotes (1 in 327), 69 compound heterozygotes, 371 simple heterozygotes, and 4,755 normals. There were 5 men and 11 women homozygotes with a mean age of 42, range 28 to 57. Mean UIBC (24 +/- 7 microL) and TS (48% +/- 17%) in homozygotes were significantly different from compound heterozygotes, simple heterozygotes, and normals (ANOVA). Only 3 homozygotes had an elevated serum ferritin. Family studies found an additional 4 iron-loaded homozygotes. Optimal thresholds were < or =28 micromol/L for UIBC and > or =46% for TS. Receiver operating characteristic (ROC) curve analysis showed an area under the curve for UIBC of 0.93 (0. 85-1.0, 95% confidence interval), and for TS of 0.83 (0.7-0.95). Screening with UIBC to preselect those for genotyping is a cost efficient strategy for population screening for hemochromatosis. PMID- 10796895 TI - Reactivity of 13 in vitro expressed hepatitis B surface antigen variants in 7 commercial diagnostic assays. AB - The primary marker of current hepatitis B infection is the surface antigen (HBsAg), however HBsAg negativity does not exclude hepatitis B viremia. HBsAg variants can be responsible for such diagnostic failures. Here 13 different HBsAg variants were cloned, variant protein produced in a mammalian expression system, and tested using 7 commercial HBsAg diagnostic assays. Of 12 variants analyzed, 6 samples displayed similar reactivity to the positive control (containing standard HBsAg sequence) in most of the assays, but 6 samples, containing various mutations throughout the entire major hydrophilic region (MHR), showed reduced reactivity. It was found that the loss of cysteine at amino acid (aa) 124 in 1 sample affected the secretion as well as the reactivity of HBsAg in the expression system. Thus, not all assays are equally able to detect HBsAg variants, implying that, to attain an acceptable level of sensitivity, the antibody repertoire of the current assays should be extended. PMID- 10796894 TI - Treatment of chronic woodchuck hepatitis virus infection in the Eastern woodchuck (Marmota monax) with nucleoside analogues is predictive of therapy for chronic hepatitis B virus infection in humans. AB - The woodchuck hepatitis virus (WHV) and its natural host, the Eastern woodchuck (Marmota monax), have been established as a model of hepatitis B virus (HBV) induced disease. Several published studies have used this experimental animal model system to demonstrate potential antiviral therapies for chronic HBV infections. However, there has been little comparative information available on compounds used in clinical anti-HBV studies in WHV-infected woodchucks, thereby making interpretations of the potential relative effectiveness of new antiviral agents in humans more difficult. In this report, using a series of placebo controlled studies, we compared the relative effectiveness of several nucleoside analogues that have been used in clinical trials for the treatment of chronic HBV infection against WHV replication in chronically infected woodchucks. Adenine-5' arabinoside monophosphate (Ara-AMP [vidarabine]), ribavirin, (-)beta-L-2',3' dideoxy-3'-thiacytidine (3TC [lamivudine]), and famciclovir (oral prodrug of penciclovir) induced depressions in viremia and intrahepatic WHV-DNA replication that were consistent with their relative effectiveness in anti-HBV human clinical trials. As observed in HBV-infected patients, 3' azido-3'-deoxythymidine (AZT [zidovudine]) had no effect on WHV replication in these studies. These experimental results more firmly establish chronic WHV infection in woodchucks as an accurate and predictive model for antiviral therapies against chronic HBV infection in humans and provide a baseline for comparative antiviral effects of other experimental antiviral agents in the WHV/woodchuck model system. PMID- 10796896 TI - Intragraft localization of activated nuclear factor kappaB in recurrent hepatitis C virus disease following liver transplantation. AB - Nuclear factor kappaB (NF-kappaB) is activated during viral infection and is central to the regulation of host immune responses. The NF-kappaB activation status and its morphological sources were assessed by immunohistochemistry in allograft biopsy specimens of orthotopic liver transplantation patients with recurrent hepatitis C virus (HCV). Hepatocellular NF-kappaB immunostaining was detected in HCV cases compared with controls (nontransplant: P <.001; transplant: P =.006), which correlated with the number of NF-kappaB positive hepatocytes (P =.007) and contrasted to the absent to weak staining of controls (nontransplant: P =.001; transplant: P =.009). Enhanced NF-kappaB staining of cytokeratin 19 positive bile ducts and proliferating ductules in the HCV group was in contrast to controls. Intense NF-kappaB immunoreactivity was detected in CD68-positive Kupffer cells and macrophages of all HCV specimens compared with a few controls (nontransplant: P <.001; transplant: P =.001) and contrasted to the weak staining of controls (nontransplant: P <.001; transplant: P =.001). NF-kappaB-positive immunoreactivity correlated with the number of T cell receptor (TCR) alpha/beta positive lymphocytes (P <.001), which was not observed in controls. In those HCV cases showing evidence of necroinflammatory activity (grade) and individual features of portal inflammation, periportal inflammation/piecemeal necrosis, lobular inflammation, and fibrosis (stage), higher NF-kappaB staining intensity scores within bile ducts, proliferating ductules, hepatocytes (piecemeal necrosis: P =.016; stage: P =.030), and lymphocytes (stage: P =.044) and increased number of NF-kappaB-positive cells within bile ducts, proliferating ductules (grade, lobular inflammation, piecemeal necrosis, stage: P =.022), hepatocytes, and lymphocytes were observed. Increased staining intensity and frequency of NF-kappaB-positive cells were similarly observed in HCV-positive allografts obtained from patients under tacrolimus- compared with cyclosporine based immunosuppression. These data implicate an immunoregulatory role of intragraft NF-kappaB activation in the pathogenesis and progression of posttransplantation HCV disease recurrence. PMID- 10796897 TI - Screening for hemochromatosis. PMID- 10796898 TI - Autoimmune hepatitis: the investigational and clinical challenges. PMID- 10796899 TI - UROTENSIN II: BETTER THAN SOMATOSTATIN FOR PORTAL HYPERTENSION? PMID- 10796900 TI - Delmar J. Stauffer RSNA executive director: a tribute. PMID- 10796901 TI - Genitourinary imaging: the past 40 years. AB - During the past 40 years, there has been a dramatic evolution in genitourinary imaging. This evolution has resulted in fundamental changes in the subspecialty. Uroradiology initially focused on radiographic imaging of the urinary tract and was practiced primarily by urologists. After the development of safe intravenous contrast materials, radiologists who focused on the urinary tract and worked closely with urologists forged major advances in urinary tract imaging and intervention. More recently, imaging of the extraurinary genital organs has been added to the subspecialty. Cross-sectional imaging techniques have supplanted radiographic imaging for both urinary and genital imaging. The emergence of the cross-sectional techniques, however, has blurred the traditional organ system based distinction between gastrointestinal radiology and genitourinary radiology, as both organ systems are imaged simultaneously, and has resulted in a new amalgamation, abdominal radiology, with roots in both specialties. The challenge for the new generation of abdominal radiologists, trained predominantly in cross sectional techniques, will be to maintain the close interaction with our clinical colleagues that the traditional organ system orientation fostered. PMID- 10796902 TI - CT scan negative for pulmonary embolism: where do we go from here? PMID- 10796903 TI - Screening for colorectal cancer. AB - Colorectal cancer is the third most commonly diagnosed cancer and the second leading cause of cancer deaths in the United States. Fortunately, both the incidence and mortality associated with the disease have declined during the past 2 decades. This is likely due, at least in part, to improved efforts at screening and more aggressive removal of adenomatous polyps. However, colorectal cancer screening is still generally underutilized. This article reviews the current status and future outlook for colorectal cancer screening, including a discussion of risk factors for the disease, its anatomic distribution, proposed mechanisms of development from adenomatous polyps, rationale for screening, and screening options. Published literature concerning the cost-effectiveness of colorectal cancer screening is also summarized. The article concludes with a discussion of the emerging consensus regarding the importance of and approaches to screening. PMID- 10796904 TI - Award-winning Papers and Honored Lectures. PMID- 10796905 TI - Appendicitis at the millennium. AB - Acute appendicitis is a common clinical problem. Accurate and prompt diagnosis is essential to minimize morbidity. While the clinical diagnosis may be straightforward in patients who present with classic signs and symptoms, atypical presentations may result in diagnostic confusion and delay in treatment. Helical computed tomography (CT) and graded compression color Doppler ultrasonography (US) are highly accurate means of establishing the diagnosis. These imaging modalities have now assumed critical roles in the treatment of patients suspected to have appendicitis. The purpose of this article is threefold: to provide an update on new information regarding the pathophysiology, clinical diagnosis, and laparoscopic treatment of acute appendicitis; to describe the state-of-the art use of CT and US in diagnosing this disease entity; and to address the role of medical imaging in this patient population. PMID- 10796906 TI - Spontaneously resolving appendicitis: frequency and natural history in 60 patients. AB - PURPOSE: To establish the frequency and natural history of ultrasonographically (US) documented spontaneously resolving appendicitis following conservative treatment. MATERIALS AND METHODS: From July 1987 to July 1997, the authors encountered 106 patients with US-diagnosed spontaneously resolving appendicitis. We retrospectively studied clinical data and US findings obtained at admission and follow-up relating to 60 patients who were treated conservatively. Over the same 10 years, 1,280 appendectomies for acute appendicitis were performed in the authors' hospital. RESULTS: Of 60 patients, 23 (38%) had recurrent appendicitis after a median of 14 weeks (range, 2-254 weeks), with 16 (70%) having recurrence within 1 year of the first attack. US findings indicated that patients with an appendiceal diameter of at least 8 mm were more prone to recurrence than patients with an appendiceal diameter of less than 8 mm; the recurrence rates were 47% (21 of 45 patients) and 13% (two of 15 patients). The other parameters did not show a statistically significant difference. CONCLUSION: Spontaneously resolving appendicitis occurs in at least one in 13 cases of appendicitis and has an overall recurrence rate of 38%, with the majority of cases reccurring within 1 year. PMID- 10796907 TI - Incidental extracolonic findings at CT colonography. AB - PURPOSE: To determine the frequency of extracolonic findings at computed tomographic (CT) colonography and the effect of these findings on subsequent patient treatment and cost. MATERIALS AND METHODS: Conventional transverse CT colonographic scans in 264 consecutive patients were evaluated independently by two radiologists. Extracolonic findings were classified as having high, moderate, or low clinical importance. The effect of CT findings on patient treatment was assessed with chart review. The cost of additional examinations was calculated by using 1999 Medicare reimbursements. RESULTS: Thirty (11%) patients had highly important extracolonic findings, which resulted in further examinations in 18 (7%) patients, including ultrasonography in 10, CT in 13, and intravenous pyelography in one. Six patients underwent surgery because of incidentally discovered CT colonographic findings. Two patients with findings of moderate or low importance underwent additional imaging. A total of $7,324 was required for work-up for extracolonic findings (mean of an additional $28 per examination). Three extracolonic malignancies were overlooked at CT colonography. CONCLUSION: Additional work-up of extracolonic CT colonographic findings was relatively infrequent but was often worthwhile when performed for lesions classified as highly important. The evaluation of extracolonic structures at CT colonography has definite limitations with regard to solid organs but can help detect serious disease without substantially increasing the cost per patient. PMID- 10796909 TI - Hepatic MR imaging with a dynamic contrast-enhanced isotropic volumetric interpolated breath-hold examination: feasibility, reproducibility, and technical quality. AB - PURPOSE: To evaluate the feasibility, reproducibility, and technical quality of a dynamic contrast material-enhanced isotropic three-dimensional (3D) volumetric interpolated breath-hold hepatic magnetic resonance (MR) imaging examination. MATERIALS AND METHODS: Fifty patients underwent 3D spoiled gradient-echo imaging (4.2/1.8 [repetition time msec/echo time msec]; flip angle, 12 degrees; interpolation in three directions; intermittent fat saturation; pixel size or =65 years (OR = 4.0, CI = 1.3-10.5, p = 0.01), peripheral vascular disease (OR = 3.4, CI = 1.3-8.8, p = 0.02), hypertension (OR = 3.0, CI = 1.2-7.0, p = 0.02), and female gender (OR = 3.0, CI = 1.2-7.1, p = 0.04). A second conservative analysis for missing data demonstrated a significant association for ACS of 80-90% alone (OR = 13.1, CI = 1.5-60.9, p = 0.01). This association remained significant after multivariate adjustment with propensity score stratification. ACS (80-90%) appears to be independently associated with ipsilateral hemispheric stroke following CS when evaluated against the present study variables. This finding supports the need for a properly conducted prospective natural history study, including an evaluation of aortic arch atherosclerosis, to determine the clinical relevance of this observation. PMID- 10796951 TI - Estimating the contemporary in-hospital costs of carotid endarterectomy. AB - Carotid endarterectomy (CEA) is the treatment of choice for symptomatic carotid stenosis and selective asymptomatic lesions. Alternative approaches have recently been championed under the guise of increased efficacy and decreased cost. The purpose of this study was to determine the results and in-hospital costs of CEA in a university hospital in the modern era. A retrospective chart review was undertaken for all patients undergoing CEA between January 1995 and December 1997. This corresponded to the implementation of a clinical path and extended efforts toward cost reduction. Patients undergoing combined CEA and cardiopulmonary bypass were excluded (n = 3). Cost was analyzed by the hospital Office of Program Planning using TSI (Transition Systems, Inc.) software. Direct costs are related to the utilization of clinical resources and are therefore manageable by clinicians (bed, room, supplies, nursing staff, OR staff, radiology, pharmacy, etc.). Total costs additionally include administration and overhead costs not directly chargeable to patient accounts. The results of this study showed that CEA can be safely performed with brief hospital stays and reasonable hospital costs. Results of alternative interventions for the treatment of carotid stenosis should be compared to these contemporary data. PMID- 10796952 TI - Major lower extremity amputation in Veterans Affairs medical centers. AB - Our objective was to assess outcomes for 8696 patients who underwent 9236 above- (AKA) and/or below-knee (BKA) amputations during a 4-year period for disorders of the circulatory system. Veterans Affairs (VA) Patient Treatment File (PTF) data were acquired for all patients in Diagnosis Related Groups (DRGs) 113 and 114 hospitalized in VA medical centers (VAMCs) during fiscal years 1991-1994. Data were further analyzed by Patient Management Category (PMC) software, which measured illness severity, patient complexity, and relative intensity score (RIS), a measure of resource utilization. The results of this analysis showed that mortality and morbidity rates remain high after AKA and BKA. Differing amputation practice patterns found in this study warrant further investigation. PMID- 10796953 TI - Splenic artery aneurysm in the 1990s. AB - Splenic artery aneurysms (SAA), although rare, are the most common visceral artery aneurysms and are known for their potential for rupture. Pregnancy and portal hypertension have been known as major risk factors. With improved methods of diagnosis and minimally invasive therapy, management and outcome of SAA may change significantly. The purpose of this study was to analyze our institutional experience with SAA during the past decade. Charts of all patients (six women, three men; mean age, 60.5 [range: 31 to 81] years) with diagnoses of SAA from 1988 to 1999 were reviewed. Associated conditions included essential hypertension (6), portal hypertension (3), diabetes (1), intracranial aneurysm (1), and polyarteritis nodosa (1). Six patients were asymptomatic, and three had ruptured SAA. Diagnosis was made by angiography (2), computed tomography (3), ultrasonography (3), and exploratory laparotomy (1). Six patients underwent surgery (five required splenectomy), one had embolization, and two had no intervention. Three postoperative deaths occurred-two (intracranial aneurysm, myocardial infarction) in the first month, one (sepsis) in the ninth month. An association of liver disease with SAA was confirmed; however, no association with pregnancy was noted. Surgical treatment followed traditional methods, and mortality correlated with presence of severe comorbidity. PMID- 10796954 TI - Endovascular grafts in the treatment of thoracic aortic aneurysms and pseudoaneurysms. AB - The purpose of this study was to describe our experience with balloon and self expanding endovascular grafts for the management of thoracic aortic lesions. Between February 1997 and June 1998, 20 endovascular grafts were implanted in 14 patients for the treatment of thoracic aortic aneurysms and pseudoaneurysms. Endovascular procedures were performed using one of four different devices: (1) Dacron-covered balloon-expandable Palmaztrade mark stent, (2) balloon-expandable Palmaz stent-PTFE graft prosthesis (BE-PS), (3) self-expanding internally supported Nitinol Dacron prosthesis (Vanguardtrade mark SE-V), and (4) self expanding externally supported Nitinol PTFE prosthesis (Excludertrade mark SE-E). The results show that endovascular grafting represents a potentially important alternative therapy to open repair of the thoracic aorta. Self-expanding devices were, in our experience, easier to use and more accurately deployed. PMID- 10796955 TI - Infected postcarotid endarterectomy pseudoaneurysms: retrospective review of a series. AB - The development of infected pseudoaneurysms (PAs) following carotid endarterectomy (CEA) is extremely rare. We retrospectively reviewed the data from 13 such aneurysms (in 12 patients) repaired over a period of 35 years at the Texas Heart Institute (THI) and included an overview of published cases to analyze the epidemiology, mode of presentation, methods of repair, and outcome. The diagnosis of PA should be considered when a patient develops a persistent hematoma, recurrent bleeding from the wound, or late wound infection. Sepsis is occasionally a presenting symptom. Surgical therapy for infected PAs can be challenging; excision of the aneurysm followed by autologous grafting constitutes the favored approach. Traditionally, Dacron patch repair has been associated with a high incidence of reinfection. Carotid artery ligation is required in a large percentage of cases (22.7%) and is associated with a prohibitive rate of death/major stroke (50%) compared with a low (12%) risk following arterial reconstruction. Prevention and early diagnosis of infected PAs are essential to limit complications and mortality. PMID- 10796956 TI - The value of the oblique groin incision for femoral artery access during endovascular procedures. AB - Groin incisions for access to femoral vessels are typically made in a vertical fashion extending across the groin crease. Significant morbidity can be associated with these incisions, including lymphoceles, lymph fistulae and infections, as documented in the infrainguinal revascularization literature. We have adopted an oblique groin incision for femoral artery access during endovascular graft reconstruction of the aorta because of the potential for reduced wound morbidity. In this study we report our experience with this technique and compare it with the existing literature to determine its usefulness. From June 1998 to May 1999, 98 consecutive patients received endovascular exclusion of aortic aneurysms at The Mount Sinai Medical Center, New York. Patients were treated with aortoaortic (24), aortouniiliac with femorofemoral crossover bypass (41), or bifurcated endografts (33) and were prospectively studied for wound complications. Aortoaortic procedures required one inguinal incision whereas aortouniiliac with femorofemoral crossover bypass and bifurcated procedures employed bilateral inguinal wounds. Wound complications were defined as cellulitis, subcutaneous purulence, femorofemoral graft infection, lymphocele, or lymphocutaneous fistulae. The oblique groin incision allows adequate exposure to the femoral arteries and is associated with low wound morbidity. We suggest that this approach may be the preferred technique for access to femoral arteries during endovascular procedures, and should be considered for infrainguinal arterial reconstructions. PMID- 10796957 TI - Percutaneous induced thrombosis of iatrogenic femoral pseudoaneurysms following catheterization. AB - Femoral pseudoaneurysms are one of the common iatrogenic complications following catheterization procedures done via the femoral approach. Their treatment has evolved over the last decade from operative repair to ultrasound-guided compression (USGC) and more recently to thrombin injection of the pseudoaneurysm. We report our experience with that technique and compare the results of thromboobliteration to those of the compression method. All consecutive iatrogenic femoral pseudoaneurysms diagnosed in the vascular laboratory of two large community hospitals were referred for the study. Under ultrasound guidance, percutaneous thromboobliteration (PTO) of the pseudoaneurysms was done by injecting thrombin solution (500-unit increments) into the pseudoaneurysm. Time to thrombosis, dose of thrombin, patient's discomfort, and ease of procedure were recorded and analyzed. Comparison with results of USGC reported in the literature was made. Percutaneous thromboobliteration may be a simple and very effective treatment of femoral pseudoaneurysms. The high success rate, ease of procedure, and cost benefit over USGC are noteworthy. A larger trial is currently under way. If results are duplicated, PTO will emerge as the preferred treatment for iatrogenic femoral pseudoaneurysms. PMID- 10796958 TI - The response of adult human saphenous vein endothelial cells to combined pressurized pulsatile flow and cyclic strain, in vitro. AB - Adult human saphenous vein endothelial cells (HVEC) were cultured in a compliant tubular device and evaluated by Northern hybridization for the effects of combined pressurized pulsatile flow and cyclic strain on the expression of mRNAs for endothelin-1 (ET-1), endothelial cell nitric oxide synthase (ecNOS), tissue plasminogen activator (tPA), and plasminogen activator inhibitor type 1 (PAI-1). The hemodynamic environment was designed to mimic shear stress conditions at the distal anastomosis of a saphenous vein graft, a common site of intimal proliferation. Steady-state mRNA levels in experimental tubes were expressed relative to that in controls. No changes were observed in ET-1 mRNA after 1 and 24 hr, but a 50% decrease in experimental cultures was observed after 48 hr in the vascular simulating device. Similar results were obtained for ecNOS mRNA, although a subgroup (4 of 11) showed a significant decrease (>50%) by 24 hr. For tPA mRNA, no change was observed after 1 hr, but a significant decrease (>60%) was measured after 24 hr and no message was detectable after 48 hr. Steady-state levels for PAI-1 mRNA remained unchanged through 48 hr of treatment. These results show that pressure, pulsatile flow, and cyclic strain, when applied in concert, differentially alter vasoactive and fibrinolytic functions in HVEC. Moreover, the dramatic decrease in steady-state levels of tPA mRNA is consistent with a shift toward an increased thrombotic state. PMID- 10796959 TI - Hyperhomocysteinemia as a cause of superior vena cava syndrome. AB - A case is presented in which superior vena cava (SVC) syndrome was caused by a stenosis of the SVC due to thrombosis. Hyperhomocysteinemia was diagnosed as a possible underlying mechanism. The role of hyperhomocysteinemia as a risk factor for the development of recurrent venous thrombosis, its diagnosis, and treatment are discussed. PMID- 10796961 TI - Rupture of the abdominal aorta in patients with Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS) is a heterogeneous inherited disorder of collagen synthesis. Type IV is frequently associated with major vascular catastrophes and challenges the vascular surgeon with its varied clinical presentation and the difficulty of vascular repair. Rupture of the abdominal aorta is one of the most serious complications and is associated with nearly 100% mortality rate. We describe here three patients with type IV EDS. PMID- 10796960 TI - Primary aortoduodenal fistula and Q fever: an underrecognized association? AB - We report a rare case of primary aortoduodenal fistula (ADF) secondary to a Coxiella burnetii (Q fever) infection in a patient with an abdominal aortic aneurysm. A review of the available literature on the vascular complications of Q fever is presented. Q fever should be suspected in vascular patients with close animal contact when a standard infectious work-up is unrevealing. Diagnostic steps and management strategies for primary ADF are also briefly reviewed. PMID- 10796962 TI - Superior mesenteric vein thrombosis and duodenal diverticulum. AB - We report a case of superior mesenteric vein thrombosis resulting from an inflamed duodenal diverticulum with associated inflammation of the uncinate process of the pancreas. PMID- 10796963 TI - Superior vena cava thrombosis after in vitro fertilization: case report and review of the literature. AB - Ovarian hyperstimulation syndrome (OHSS) is a rare complication of fertility medication. A 33 year old female with OHSS with thrombosis of the right internal jugular vein, subclavian vein, and superior vein cava underwent in vitro fertilization following stimulation with a GnRH analog with successful implantation. The patient developed abdominal distention and dyspnea, with persistent symptoms that resulted in a 20 lb weight loss. As pregnancy progressed, edema, pain, and tingling sensations developed by the ninth week at which time a CT scan confirmed thrombus with the right internal jugular and subclavian vein and a free floating tip in the superior vena cava. Following treatment with intravenous heparin therapy and subcutaneous low-molecular weight heparin until delivery her symptoms improved. While optimal treatment remains unclear, treatment strategies remain conservative. Identifying the risk factors that lead to the development of OHSS including the identification of those patients at risk for developing OHSS, more extensive investigation of potential underlying coagulopathy in severe or recurrent cases, and consideration of prophylactic subcutaneous heparin or IV albumin supplementation, will facilitate prevention in the high risk population. PMID- 10796964 TI - Popliteal venous aneurysms: report of an unusual presentation and literature review. AB - Aneurysms of the popliteal vein represent a rare clinical finding. Most patients with popliteal venous aneurysms present with pulmonary emboli. We describe a case report of a 69-year-old female who presented with popliteal fossa pain. A popliteal venous aneurysm was detected following extensive diagnostic work-up that included magnetic resonance imaging (MRI), venous duplex, and venography. At operation, the aneurysm was found to be compressing the tibial nerve. Tangential aneurysmectomy and lateral venorrhaphy was performed. A PTFE sleeve was placed around the site of venous repair to prevent recurrence of the aneurysm and to keep the tibial nerve free of the repair site. The postoperative course was uneventful, and duplex studies at 18 months after the procedure remain normal. The etiology of popliteal venous aneurysms is discussed here, and presentation, diagnostic work-up, and treatment options are reviewed. We demonstrate that pain in the popliteal fossa should be considered a presenting symptom for venous aneurysms. PMID- 10796965 TI - Endovascular femoropopliteal bypass. PMID- 10796966 TI - Axillobifemoral bypass. PMID- 10796967 TI - Progress in protecting reproductive rights and promoting reproductive health: five years since Cairo. PMID- 10796969 TI - Reproductive rights: how to move forward? PMID- 10796968 TI - Rights and needs: rethinking the connections in debates over reproductive and sexual rights. PMID- 10796970 TI - Rights to sexual and reproductive health: the ICPD and the convention on the elimination of all forms of discrimination against women. AB - This article examines women's rights to sexual and reproductive health as recognized by the ICPD and expressed in the Women's Convention and other international human rights documents. Rights relating to reproductive and sexual health include the rights to life, liberty, and the security of the person; to health care and information; and to nondiscrimination in the allocation of resources to health services and in their availability and accessibility. Of central importance are the rights to autonomy and privacy in making sexual and reproductive decisions, as well as the responsibility of health care providers to ensure informed consent and confidentiality in relation to health services. The article presents country examples from States parties' periodic reports under the Women's Convention that reflect systemic violations of the above rights in varied forms. PMID- 10796971 TI - Sexual but not reproductive: exploring the junction and disjunction of sexual and reproductive rights. AB - Although the term "sexual rights" has gained widespread currency, its concrete scope and content have not yet been fully defined. The need for definition is critical not only for promoting governmental accountability but also for ensuring that sexual rights can be claimed by diverse persons around the world. Ironically, the concept of "sexual and reproductive rights" poses a challenge to this effort; practices and people not traditionally addressed by reproductive rights work must be explicitly named and protected. This article considers how international norms have contributed to a gendered regulation of sexuality and of contemporary theories of "socially constructed sexuality," and it proposes a focus on the conditions that contribute to the ability to choose and on the links between sexuality, conduct, identity, social structures, and reproduction. Given the probable politically charged responses, global coalition-building is needed. PMID- 10796972 TI - The "double discourse" on sexual and reproductive rights in Latin America: the chasm between public policy and private actions. AB - This article examines how political controversies affect citizens' ability to exercise sexual and reproductive rights in Latin America. The article argues that societies accommodate conflicting views on sexuality and reproduction with a "double discourse system," which defends repressive or negligent public policies while privately tolerating unofficial and often illegal mechanisms that expand private sexual and reproductive choices. The examples of divorce policy in Chile and abortion policy in Colombia and Chile are highlighted to illustrate how this breach between public discourse and private actions operates in practice, and who is harmed by it. The article concludes by discussing the implications of this system for rights advocacy. PMID- 10796973 TI - Sexual rights in southern Africa: a Beijing discourse or a strategic necessity? AB - This article explores the meaning of sexual rights as interpreted by different stakeholders during the development of the Beijing Programme of Action and within the Southern African Development Community (SADC). It illustrates how the lack of sexual rights as understood in the African context results from poverty as well as gender inequality, particularly in sexual relationships. This lack is manifested in the circumstances surrounding the HIV/AIDS pandemic and violence against women. In the European context, in contrast, sexual rights claims are motivated specifically in relation to sexual orientation. The article explores the extent to which these different discourses are being addressed in practice in SADC member countries and the opportunities that exist for building a concrete practice of sexual rights both in the region and internationally. PMID- 10796974 TI - Abortion in the United States: barriers to access. AB - The legalization of abortion in the United States has brought a dramatic improvement in women's health and reductions in maternal and infant mortality. For young women, low-income women, and women of color, however, access to abortion has been increasingly restricted. This article describes the obstacles to abortion access, including lack of federal funding; restrictive laws, encompassing those requiring parental consent or notification for a minor seeking an abortion, as well as those attempting to ban a certain procedure; stigmatization and marginalization of abortion; decreasing abortion services; and a shortage of providers. The article connects the erosions in rights relating to abortion to policies undermining poor women's rights in relation to having children. PMID- 10796975 TI - Reproductive rights in Hungarian law: a new right to assisted procreation? AB - Hungary has a mixed record in terms of fulfilling reproductive rights as a whole, but in the context of artificially assisted procreation, it provides reproductive health services far beyond those offered by its neighbors, beyond what is stipulated by the ICPD Programme of Action, and, arguably, beyond the internationally accepted parameters of reproductive rights. Recent legislation on assisted procreation has established important new regulations and formulated a new "right to continuation of infertility treatment" applicable to women who have been widowed or divorced. The new legislation is examined in the context of the international reproductive rights movement, with comparisons to other European countries and with reference to Hungarian attitudes and laws on abortion and surrogacy. PMID- 10796976 TI - The Amazonian Peoples' Resources Initiative: promoting reproductive rights and community development in the Peruvian Amazon. PMID- 10796977 TI - Operationalizing Cairo and Beijing: a training initiative in gender and reproductive health. The International Coordinating Committee. PMID- 10796978 TI - Foreword and introduction to the book (strept)avidin-biotin system. PMID- 10796979 TI - The X-ray three-dimensional structure of avidin. AB - Avidin is a basic, highly stable, homotetrameric protein, isolated from bird egg white, binding up to four molecules of D-biotin with extremely high affinity (Kd approximately 10(-15) M). The protein has been the object of different crystallographic investigations. In all the crystal structures, the four avidin subunits display almost exact 222 symmetry. Each avidin chain (128 amino acids) is arranged in a eight-stranded antiparallel beta-barrel, whose inner region defines the D-biotin binding site. The molecular bases of D-biotin affinity can be recognised in a fairly rigid binding site, which is sterically complementary to the shape and polarity of the incoming vitamin, and is readily accessible in the apoprotein structure. Avidin displays remarkable structural and functional relationships to the acidic protein sretpavidin, isolated from Streptomyces avidinii. PMID- 10796980 TI - X-ray crystallographic studies of streptavidin mutants binding to biotin. AB - On the basis of high resolution crystallographic studies of streptavidin and its biotin complex, three principal binding motifs have been identified that contribute to the tight binding. A flexible binding loop can undergo a conformational change from an open to a closed form when biotin is bound. Additional studies described here of unbound wild-type streptavidin have provided structural views of the open conformation. Several tryptophan residues packing around the bound biotin constitute the second binding motif, one dominated by hydrophobic interactions. Mutation of these residues to alanine or phenylalanine have variable effects on the thermodynamics and kinetics of binding, but they generate only small changes in the molecular structure. Hydrogen bonding interactions also contribute significantly to the binding energetics of biotin, and the D128A mutation which breaks a hydrogen bond between the protein and a ureido NH group results in a significant structural alteration that could mimic an intermediate on the dissociation pathway. In this review, we summarize the structural aspects of biotin recognition that have been gained from crystallographic analyses of wild-type and site-directed streptavidin mutants. PMID- 10796981 TI - Two-dimensional streptavidin crystals: macropatterns and micro-organization. AB - Two dimensional crystals of streptavidin grown on lipid monolayers can be viewed as model systems for the study of phase transitions and morphology. These crystals form a variety of macroscopic morphologies associated with different microscopic crystal structures. Observed morphologies are similar to those found in two-dimensional lipid systems, and growth of the protein arrays is somewhat analogous. Such solid state physical processes as nucleation, transformation between crystal phases, crystal phase coexistence, and roughening have been observed in the streptavidin system. In this review, we highlight observations that cause streptavidin to remain an interesting model system exhibiting a variety of intriguing phenomena. PMID- 10796982 TI - Two-dimensional crystallization of streptavidin: in pursuit of the molecular origins of structure, morphology, and thermodynamics. AB - The streptavidin two-dimensional (2D) crystallization model has served as a paradigm for molecular self-assembly at interfaces. We have developed quantitative Brewster angle microscopy for the in situ measurement of spatially resolved relative protein surface densities. This allows investigation of both the thermodynamics and morphologies of 2D crystal growth. For crystal structure analysis, we employ TEM on grown crystals transferred to solid substrates. Comparison of results between commercially available streptavidin, recombinant streptavidin, and site-directed streptavidin mutants has provided insight into the protein protein and protein-lipid interactions that underlie 2D crystallization. PMID- 10796983 TI - Streptavidin-biotin binding energetics. AB - The high affinity energetics in the streptavidin-biotin system provide an excellent model system for studying how proteins balance enthalpic and entropic components to generate an impressive overall free energy for ligand binding. We review here concerted site-directed mutagenesis, biophysical, and computational studies of aromatic and hydrogen bonding interaction energetics between streptavidin and biotin. These results also have provided insight into how streptavidin builds a large activation barrier to dissociation by managing the enthalpic and entropic activation components. Finally, we review recent studies of the biotin dissociation pathway that address the fundamental question of how ligands exit protein binding pockets. PMID- 10796984 TI - Direct force measurements of the streptavidin-biotin interaction. AB - The interaction between streptavidin and its ligand, biotin, were studied by direct force measurements. The complimentary approaches of surface force apparatus (SFA) and atomic force microscopy (AFM) were used to elucidate both long-range and short-range adhesive interactions of the streptavidin biotin interaction. The high spatial resolution of the SFA provided a detailed profile of the intersurface forces of apposing surfaces functionalized with streptavidin and biotin. Measurements obtained by the SFA corresponded to long and intermediate-range forces that are important in determining ligand receptor association. AFM was used to measure the unbinding force of individual streptavidin biotin complexes. These measurements revealed the short-range interactions (i.e. hydrophobic and hydrogen bonding forces) that stabilize the intermolecular bond. PMID- 10796985 TI - Streptavidin-binding and -dimerizing ligands discovered by phage display, topochemistry, and structure-based design. AB - Structural and mechanistic determinants of affinity of streptavidin-binding peptide ligands discovered by phage display are reviewed along with the use of streptavidin as a paradigm for structure-based design. A novel way of producing protein-dimerizing ligands in the streptavidin model system is discussed, in which crystal packing topochemically mediates or even catalyzes dimerization of adjacent bound ligands whose reactive ligating groups are presented toward one another in productive orientations in the crystal lattice. Finally, through crystallography on a set of streptavidin complexes with small molecule and peptide ligands at multiple pHs in two space groups, the mechanism by which ligands enhance intersubunit stabilization of the streptavidin tetramer is probed. PMID- 10796986 TI - Extremely high thermal stability of streptavidin and avidin upon biotin binding. AB - The effect of biotin binding on the thermal stability of streptavidin (STV) and avidin (AVD) was evaluated using differential scanning calorimetry. Biotin binding increases the midpoint of temperature Tm of thermally induced denaturation of STV and AVD in phosphate buffer from 75 and 83 degrees C to 112 and 117 degrees C at full biotin saturation, respectively. This thermostability is the highest reported for proteins coming from either mesophilic or thermophilic organisms. In both proteins, biotin also increases the calorimetric enthalpy and the cooperativity of the unfolding. Thermal stability of STV was also evaluated in the presence of high concentrations of urea or guanidinium hydrochloride (GuHCl). In 6 M GuHCl, STV remains as a tetramer and the Tm of the STV-biotin complex is centered at 108 degrees C, a few degrees below the value obtained in phosphate buffer. On the contrary, STV under fully saturating condition remains mainly in its dimeric form in 8 M urea and the thermogram shows two endotherms. The main endotherm at a lower temperature has been ascribed to the dimeric liganded state with a Tm of 87 degrees C, and the higher temperature endotherm to the tetrameric liganded form with a Tm of 106 degrees C. As the thermostability of unliganded protein in the presence of urea is unchanged upon binding we related the extremely high thermal stability of this protein to both an increase in structural ordering and compactness with the preservation of the tetramer integrity. PMID- 10796987 TI - Avidin and streptavidin in clinical diagnostics. AB - With the introduction of the SA/Bi technology into the clinical diagnostic field ultrasensitive tests with an acceptable turn-around time became feasible. Cytokine ELISAs with lower detection limit in the picogram range became finally available. Prior to the introduction of SA/Bi only elevated levels of target could be detected by short time tests but not the normal base level. The turn around time for standard clinical immunological tests was reduced to 10-20 min from 2-4 h in old assays. PMID- 10796988 TI - Applications of a peptide ligand for streptavidin: the Strep-tag. AB - The Strep-tag constitutes a nine amino acid-peptide that binds specifically to streptavidin and occupies the same pocket where biotin is normally complexed. Since the Strep-tag participates in a reversible interaction it can be applied for the efficient purification of corresponding fusion proteins on affinity columns with immobilized streptavidin. Elution of the bound recombinant protein can be effected under mild buffer conditions by competition with biotin or a suitable derivative. In addition, Strep-tag fusion proteins can be easily detected in immunochemical assays, like Western blots or ELISAs, by means of commercially available streptavidin-enzyme conjugates. The Strep-tag/streptavidin system has been systematically optimized over the past years, including the engineering of streptavidin itself. Structural insight into the molecular mimicry between the peptide and biotin was furthermore gained from X-ray crystallographic analysis. As a result the system provides a reliable and versatile tool in recombinant protein chemistry. Exemplary applications of the Strep-tag are discussed in this review. PMID- 10796989 TI - Recombinant avidin and avidin-fusion proteins. AB - Both chicken egg-white avidin and its bacterial relative streptavidin are well known for their extraordinary high affinity with biotin (Kd approximately 10(-15) M). They are widely used as tools in a number of affinity-based separations, in diagnostic assays and in a variety of other applications. These methods have collectively become known as (strept)avidin-biotin technology. Biotin can easily and effectively be attached to different molecules, termed binders and probes, without destroying their biological activity. The exceptional stability of the avidin-biotin complex and the wide range of commercially available reagents explain the popularity of this system. In order by genetic engineering to modify the unwanted properties of avidin and to further expand the existing avidin biotin technology, production systems for recombinant avidin and avidin-fusion proteins have been established. This review article presents an overview of the current status of these systems. Future trends in the production and applications of recombinant avidin and avidin-fusion proteins are also discussed. PMID- 10796990 TI - Smart and biofunctional streptavidin. AB - The high affinity recognition of biotin and biotinylated molecules has made streptavidin one of the most important components in diagnostics and laboratory kits. While it is extremely useful as the native protein, there are many applications where its function can be improved re-engineering the subunits. We review here our efforts to construct streptavidin tetramers that have 'smart' recognition capabilities, and which display functional peptide sequences. These smart and biofunctional streptavidin derivatives can 'talk' to cells, and 'listen' to external signals which control capture and release of biotinylated molecules. PMID- 10796991 TI - A streptavidin-luciferase fusion protein: comparisons and applications. AB - Luciferases are unique enzymes in being capable of emitting visible light as one of the end-products of their catalysis. Both procaryotic and eucaryotic organisms exist that emit light, and the luciferases from these organisms differ considerably in size as well as chemistry of catalysis. Two main, i.e. most studied groups, are the bacterial luciferases of e.g. Vibrio fisheri, Vibrio harveyi, and Photorhabdus luminescens, responding to FMNH2, long-chain aldehyde and molecular oxygen and the insect luciferases of the fireflies Photinus pyralis and Luciola minengrelica or click beetle Pyrophorus plagiophthalamus, responding to ATP, luciferin and molecular oxygen. An emerging amount of 'new' luciferases from shrimps, fish, jelly fish and overall from marine origin, are finding their way to biotechnological applications. The common feature of these is their ability to produce light within the visible region of the spectrum, i.e. between 450 nm (blue) and 630 nm (red). In this short review, we discuss some of the recent advances on fusion proteins of eucaryotic luciferases and their applications. Special emphasis is placed on a streptavidin-luciferase fusion protein produced by insect cells using the baculovirus expression system. PMID- 10796992 TI - Molecular genetic applications of streptavidin-coated manifold supports. AB - Practical problems of handling large numbers of samples limit the application of molecular genetic procedures in clinical settings and in research. In the present review we describe a multipronged manifold support, coated with streptavidin, that offers distinct advantages in preparative and diagnostic applications. In order to increase the surface available on the manifold, porous Sepharose particles conjugated with streptavidin were attached to the plastic support. This procedure increased the surface by almost three orders of magnitude, permitting sufficient streptavidin to be coupled to the support for most routine applications. The manifold supports have been used for sample preparation and in a number of genetic assays, including allele discrimination assays and DNA sequencing, In all these assay formats the manifold supports allow large numbers of samples to be processed in parallel. PMID- 10796993 TI - Development of new biotin/streptavidin reagents for pretargeting. AB - The high affinity of biotin for streptavidin has made this pair of molecules very useful for in vivo applications. To optimize reagents for one potential in vivo application, antibody-based pretargeting of cancer, we have prepared a number of new biotin and streptavidin derivatives. The derivatives developed include new radiolabeled biotin reagents, new protein biotinylation reagents, and new biotin multimers for cross-linking and/or polymerization of streptavidin. We have also modified streptavidin by site-directed mutation and chemical modification to improve its in vivo characteristics, and have developed new reagents for cross linking antibody fragments with streptavidin. A brief overview of these new reagents is provided. PMID- 10796994 TI - In vivo enzymatic protein biotinylation. AB - Biotin is biologically active only when protein-bound and is covalently attached to a class of important metabolic enzymes, the biotin carboxylases and decarboxylases. Biotinylation is a relatively rare modification, with between one and five biotinylated protein species found in different organisms. We discuss the mechanism and structures involved in this extraordinarily specific protein modification and its exploitation in tagging recombinant proteins. PMID- 10796995 TI - Matrix-assisted laser desorption/ionization mass spectrometry of DNA using photocleavable biotin. AB - Oligonucleotides containing a photocleavable biotin (5'-PC-biotin) were analyzed by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) with wavelengths in the ultraviolet (UV) and infrared (IR) from solution and after capture on streptavidin-coated agarose or magnetic beads. The analysis was used to monitor the release of the oligonucleotides as a result of photochemical cleavage of the biotinylated linker. Near-UV pulses (UV-MALDI) led to predominant release of the photocleaved product. In contrast, only the uncleaved analyte was detected using IR pulses (IR-MALDI). Results from MALDI analysis are also presented for DNA containing a photocleavable 5'-amino group which can be covalently linked to a variety of activated surfaces and marker molecules. In a demonstration of this approach, a 5'-PC-biotinylated 49 nt RNA oligonucleotide was enzymatically synthesized using a PC-biotin-r(AG) dinucleotide primer, captured on streptavidin coated magnetic beads and analyzed by UV-MALDI. Potential applications of photocleavable linkers combined with MALDI for the analysis of nucleic acids are discussed. PMID- 10796996 TI - Streptavidin-based containment systems for genetically engineered microorganisms. AB - The use of genetically modified microorganisms for environmental remediation continues to be debated. Conditional lethal systems with tightly regulated gene expression can be used to contain released microorganisms and ameliorate some of the concerns about horizontal gene transfer. We have described streptavidin-based suicide systems to address these concerns and evaluated their function in Pseudomonas putida containing the TOL plasmid for aromatic hydrocarbon metabolism. Tight regulation of expression of a truncated streptavidin gene was required to avoid premature production of the toxic protein. Streptavidin expression was induced by the absence of 3-methyl benzoate (hydrocarbon substrate) which resulted in the elimination of 99.9% of the bacterial culture within eight hours. Low mutant escape rates at 10(-7) per cell per generation were also realized. PMID- 10796997 TI - Whither the future of controlling quantum phenomena? AB - This review puts into perspective the present state and prospects for controlling quantum phenomena in atoms and molecules. The topics considered include the nature of physical and chemical control objectives, the development of possible quantum control rules of thumb, the theoretical design of controls and their laboratory realization, quantum learning and feedback control in the laboratory, bulk media influences, and the ability to utilize coherent quantum manipulation as a means for extracting microscopic information. The preview of the field presented here suggests that important advances in the control of molecules and the capability of learning about molecular interactions may be reached through the application of emerging theoretical concepts and laboratory technologies. PMID- 10796998 TI - Micropatterning of organic electronic devices by cold-welding AB - A simple and general postdeposition electrode patterning technique for active organic electronic devices is demonstrated and is applied to patterning the metal cathodes of organic light-emitting devices. Selective lift-off of the metal cathode layer is achieved by pressing a prepatterned, metal-coated silicon stamp on the unpatterned device layers. Under pressure, the metal coating on the stamp cold-welds to the metal cathode coating the underlying organic films. Subsequent separation of the stamp from the substrate results in removal of the cathode metal in the regions contacted by the stamp, resulting in submicrometer feature definition. A 17x17 passive matrix display, with a pixel size of 440 micrometers by 320 micrometers, was fabricated with this process. Cold-welding followed by lift-off of the cathode metal allows simple, cost-effective, and high-throughput large-area fabrication of organic electronic devices. PMID- 10796999 TI - Stable versions of transient push-pull carbenes: extending lifetimes from nanoseconds to weeks AB - A (phosphanyl)(trifluoromethyl)carbene is shown to be stable for weeks in solution at temperatures up to -30 degrees C. Its chemical behavior exactly matches that of its transient congeners, even to the extent that subtle effects, such as the recently discovered weak pi interaction with aromatics, are observed. The influence of substituents on the structure of push-pull carbenes is demonstrated by a single-crystal x-ray diffraction study of a (phosphanyl)[2, 6 bis(trifluoromethyl)phenyl]carbene. The lifetime of these molecules makes them accessible to a wider range of standard techniques, allowing their chemical and physical behaviors to be studied in more detail than was previously possible. PMID- 10797000 TI - Radar observations of asteroid 216 kleopatra AB - Radar observations of the main-belt, M-class asteroid 216 Kleopatra reveal a dumbbell-shaped object with overall dimensions of 217 kilometers by 94 kilometers by 81 kilometers (+/-25%). The asteroid's surface properties are consistent with a regolith having a metallic composition and a porosity comparable to that of lunar soil. Kleopatra's shape is probably the outcome of an exotic sequence of collisional events, and much of its interior may have an unconsolidated rubble pile structure. PMID- 10797001 TI - Large-scale thermal events in the solar nebula: evidence from Fe,Ni metal grains in primitive meteorites AB - Chemical zoning patterns in some iron, nickel metal grains from CH carbonaceous chondrites imply formation at temperatures from 1370 to 1270 kelvin by condensation from a solar nebular gas cooling at a rate of approximately 0.2 kelvin per hour. This cooling rate requires a large-scale thermal event in the nebula, in contrast to the localized, transient heating events inferred for chondrule formation. In our model, mass accretion through the protoplanetary disk caused large-scale evaporation of precursor dust near its midplane inside of a few astronomical units. Gas convectively moved from the midplane to cooler regions above it, and the metal grains condensed in these parcels of rising gas. PMID- 10797002 TI - Age of Neoproterozoic bilatarian body and trace fossils, White Sea, Russia: implications for metazoan evolution. AB - A uranium-lead zircon age for a volcanic ash interstratified with fossil-bearing, shallow marine siliciclastic rocks in the Zimnie Gory section of the White Sea region indicates that a diverse assemblage of body and trace fossils occurred before 555.3 +/- 0.3 million years ago. This age is a minimum for the oldest well documented triploblastic bilaterian Kimberella. It also makes co-occurring trace fossils the oldest that are reliably dated. This determination of age implies that there is no simple relation between Ediacaran diversity and the carbon isotopic composition of Neoproterozoic seawater. PMID- 10797003 TI - 40K-(40)Ar constraints on recycling continental crust into the mantle AB - Extraction of potassium into magmas and outgassing of argon during melting constrain the relative amounts of potassium in the crust with respect to those of argon in the atmosphere. No more than 30% of the modern mass of the continents was subducted back into the mantle during Earth's history. It is estimated that 50 to 70% of the subducted sediments are reincorporated into the deep continental crust. A consequence of the limited exchange between the continental crust and the upper mantle is that the chemistry of the upper mantle is driven by exchange of material with the deep mantle. PMID- 10797004 TI - Satellite measurements of sea surface temperature through clouds AB - Measurements of sea surface temperature (SST) can be made by satellite microwave radiometry in all weather conditions except rain. Microwaves penetrate clouds with little attenuation, giving an uninterrupted view of the ocean surface. This is a distinct advantage over infrared measurements of SST, which are obstructed by clouds. Comparisons with ocean buoys show a root mean square difference of about 0.6 degrees C, which is partly due to the satellite-buoy spatial-temporal sampling mismatch and the difference between the ocean skin temperature and bulk temperature. Microwave SST retrievals provide insights in a number of areas, including tropical instability waves, marine boundary layer dynamics, and the prediction of hurricane intensity. PMID- 10797005 TI - Cooperation through image scoring in humans. AB - The "tragedy of the commons," that is, the selfish exploitation of resources in the public domain, is a reason for many of our everyday social conflicts. However, humans are often more helpful to others than evolutionary theory would predict, unless indirect reciprocity takes place and is based on image scoring (which reflects the way an individual is viewed by a group), as recently shown by game theorists. We tested this idea under conditions that control for confounding factors. Donations were more frequent to receivers who had been generous to others in earlier interactions. This shows that image scoring promotes cooperative behavior in situations where direct reciprocity is unlikely. PMID- 10797006 TI - Species diversity and biological invasions: relating local process to community pattern. AB - In a California riparian system, the most diverse natural assemblages are the most invaded by exotic plants. A direct in situ manipulation of local diversity and a seed addition experiment showed that these patterns emerge despite the intrinsic negative effects of diversity on invasions. The results suggest that species loss at small scales may reduce invasion resistance. At community-wide scales, the overwhelming effects of ecological factors spatially covarying with diversity, such as propagule supply, make the most diverse communities most likely to be invaded. PMID- 10797007 TI - Homogenization of fish faunas across the United States. AB - Fish faunas across the continental United States have become more similar through time because of widespread introductions of a group of cosmopolitan species intended to enhance food and sport fisheries. On average, pairs of states have 15.4 more species in common now than before European settlement of North America. The 89 pairs of states that formerly had no species in common now share an average of 25.2 species. Introductions have played a larger role than extirpations in homogenizing fish faunas. Western and New England states have received the most introductions, which is a reflection of the small number of native fishes in these areas considered desirable gamefish by settlers. PMID- 10797008 TI - Evidence for brainstem structures participating in oculomotor integration. AB - The cerebellar flocculus has been implicated in vestibulo-oculomotor control. One major central input to this structure originates from brainstem cells in the paramedian tract (PMT), whose function is unknown. Here it is reported that PMT cells in the pons carry vestibular and eye movement signals and their pharmacological inactivation produces a leaky integrator combined with vestibular imbalance. The results suggest that PMT cells provide the cerebellum with sensory and motor signals that are essential for velocity-to-position integration, a common premotor process that is required in all motor systems. PMID- 10797010 TI - Template boundary in a yeast telomerase specified by RNA structure. AB - The telomerase ribonucleoprotein has a phylogenetically divergent RNA subunit, which contains a short template for telomeric DNA synthesis. To understand how telomerase RNA participates in mechanistic aspects of telomere synthesis, we studied a conserved secondary structure adjacent to the template. Disruption of this structure caused DNA synthesis to proceed beyond the normal template boundary, resulting in altered telomere sequences, telomere shortening, and cellular growth defects. Compensatory mutations restored normal telomerase function. Thus, the RNA structure, rather than its sequence, specifies the template boundary. This study reveals a specific function for an RNA structure in the enzymatic action of telomerase. PMID- 10797009 TI - Direct targeting of light signals to a promoter element-bound transcription factor. AB - Light signals perceived by the phytochrome family of sensory photoreceptors are transduced to photoresponsive genes by an unknown mechanism. Here, we show that the basic helix-loop-helix transcription factor PIF3 binds specifically to a G box DNA-sequence motif present in various light-regulated gene promoters, and that phytochrome B binds reversibly to G-box-bound PIF3 specifically upon light triggered conversion of the photoreceptor to its biologically active conformer. We suggest that the phytochromes may function as integral light-switchable components of transcriptional regulator complexes, permitting continuous and immediate sensing of changes in this environmental signal directly at target gene promoters. PMID- 10797011 TI - Promoter-selective properties of the TBP-related factor TRF1. AB - The TATA-binding protein (TBP)-related factor 1 (TRF1) is expressed in a tissue restricted fashion during Drosophila embryogenesis and may serve as a promoter specific recognition factor that can replace TBP in regulating transcription. However, bona fide target promoters that would preferentially respond to TRF1 have remained elusive. Polytene chromosome staining, chromatin immunoprecipitation, direct messenger RNA analysis, and transient cotransfection assays identified the Drosophila gene tudor as containing a TRF1-responsive promoter. Reconstituted in vitro transcription reactions and deoxyribonuclease I footprinting assays confirmed the ability of TRF1 to bind preferentially and direct transcription of the tudor gene from an alternate promoter. Thus, metazoans appear to have evolved gene-selective and tissue-specific components of the core transcription machinery to regulate gene expression. PMID- 10797012 TI - Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. AB - The c-Jun NH2-terminal kinase (JNK) is activated when cells are exposed to ultraviolet (UV) radiation. However, the functional consequence of JNK activation in UV-irradiated cells has not been established. It is shown here that JNK is required for UV-induced apoptosis in primary murine embryonic fibroblasts. Fibroblasts with simultaneous targeted disruptions of all the functional Jnk genes were protected against UV-stimulated apoptosis. The absence of JNK caused a defect in the mitochondrial death signaling pathway, including the failure to release cytochrome c. These data indicate that mitochondria are influenced by proapoptotic signal transduction through the JNK pathway. PMID- 10797013 TI - Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli. AB - To determine why proteasome inhibitors prevent thymocyte death, we examined whether proteasomes degrade anti-apoptotic molecules in cells induced to undergo apoptosis. The c-IAP1 and XIAP inhibitors of apoptosis were selectively lost in glucocorticoid- or etoposide-treated thymocytes in a proteasome-dependent manner before death. IAPs catalyzed their own ubiquitination in vitro, an activity requiring the RING domain. Overexpressed wild-type c-IAP1, but not a RING domain mutant, was spontaneously ubiquitinated and degraded, and stably expressed XIAP lacking the RING domain was relatively resistant to apoptosis-induced degradation and, correspondingly, more effective at preventing apoptosis than wild-type XIAP. Autoubiquitination and degradation of IAPs may be a key event in the apoptotic program. PMID- 10797014 TI - Seeing the herpesvirus capsid at 8.5 A. AB - Human herpesviruses are large and structurally complex viruses that cause a variety of diseases. The three-dimensional structure of the herpesvirus capsid has been determined at 8.5 angstrom resolution by electron cryomicroscopy. More than 30 putative alpha helices were identified in the four proteins that make up the 0.2 billion-dalton shell. Some of these helices are located at domains that undergo conformational changes during capsid assembly and DNA packaging. The unique spatial arrangement of the heterotrimer at the local threefold positions accounts for the asymmetric interactions with adjacent capsid components and the unusual co-dependent folding of its subunits. PMID- 10797015 TI - Anticoagulation for patients with atrial fibrillation and risk factors for stroke. Warfarin reduces the risk by two thirds, but doctors still aren't prescribing it enough. PMID- 10797016 TI - How to improve communication between doctors and patients. Learning more about the decision making context is important. PMID- 10797017 TI - Treating hyperhidrosis. Surgery and botulinum toxin are treatments of choice in severe cases. PMID- 10797018 TI - Waiting for the working draft from the human genome project. A huge achievement, but not of immediate medical use. PMID- 10797019 TI - Junior doctors' pay: a wake-up call to the NHS. The new pay deal will also improve doctors' and patients' health and safety. PMID- 10797020 TI - Gene therapy saves immune deficient babies in France. PMID- 10797021 TI - US plans drugs stockpile to counter bioterrorism threat. PMID- 10797022 TI - Cancer "tsar" invites ideas for modernization. PMID- 10797023 TI - First results of vCJD survey show no signs of prion. PMID- 10797024 TI - In brief PMID- 10797026 TI - Britain's first "virtual hospital" gets go ahead PMID- 10797025 TI - Dutch workers entitled to smoke-free conditions, court rules. PMID- 10797027 TI - Ireland's junior doctors to strike PMID- 10797028 TI - African heads of state promise action against malaria. PMID- 10797029 TI - Research does not reflect global disease burden. PMID- 10797030 TI - Risk factors for neonatal encephalopathy in Kathmandu, Nepal, a developing country: unmatched case-control study. AB - OBJECTIVE: To determine the risk factors for neonatal encephalopathy among term infants in a developing country. DESIGN: Unmatched case-control study. SETTING: Principal maternity hospital of Kathmandu, Nepal. SUBJECTS: All 131 infants with neonatal encephalopathy from a population of 21 609 infants born over an 18 month period, and 635 unmatched infants systematically recruited over 12 months. MAIN OUTCOME MEASURES: Adjusted odds ratio estimates for antepartum and intrapartum risk factors. RESULTS: The prevalence of neonatal encephalopathy was 6.1 per 1,000 live births of which 63% were infants with moderate or severe encephalopathy. The risk of death from neonatal encephalopathy was 31%. The risk of neonatal encephalopathy increased with increasing maternal age and decreasing maternal height. Antepartum risk factors included primiparity (odds ratio 2.0) and non-attendance for antenatal care (2.1). Multiple births were at greatly increased risk (22). Intrapartum risk factors included non-cephalic presentation (3.4), prolonged rupture of membranes (3.8), and various other complications. Particulate meconium was strongly associated with encephalopathy (18). Induction of labour with oxytocin was associated with encephalopathy in 12 of 41 deliveries (5.7). Overall, 78 affected infants (60%) compared with 36 controls (6%) either had evidence of intrapartum compromise or were born after an intrapartum difficulty likely to result in fetal compromise. A concentration of maternal haemoglobin of less than 8.0 g/dl in the puerperium was significantly associated with encephalopathy (2.5) as was a maternal thyroid stimulating hormone concentration greater than 5 mIU/l (2.1). CONCLUSIONS: Intrapartum risk factors remain important for neonatal encephalopathy in developing countries. There is some evidence of a protective effect from antenatal care. The use of oxytocin in low income countries where intrapartum monitoring is suboptimal presents a major risk to the fetus. More work is required to explore the association between maternal deficiency states and neonatal encephalopathy. PMID- 10797031 TI - Prospective cohort study to determine if trial efficacy of anticoagulation for stroke prevention in atrial fibrillation translates into clinical effectiveness. AB - OBJECTIVE: To determine whether trial efficacy of prophylaxis with warfarin for patients with atrial fibrillation at high risk of stroke translates into effectiveness in clinical practice. DESIGN: Two year prospective cohort study. SETTING: District general hospital. PARTICIPANTS: 167 patients with atrial fibrillation and at high stroke risk who were eligible for anticoagulation. INTERVENTIONS: Long term anticoagulation with warfarin at adjusted doses to maintain an international normalised ratio of 2.0-3.0. MAIN OUTCOME MEASURES: Comparison of patient characteristics, comorbidity, anticoagulation control, stroke rate, and haemorrhagic complications with pooled data from five randomised controlled trials. RESULTS: Patients in the study group were seven years older (95% confidence interval 4 to 10) and comprised 33% more women than patients in the pooled trials. The international normalised ratio was in the target range for 61% of the time (range 37%-85%), below for 26% of the time (range 8%-32%), and above for 13% of the time (range 6%-26%). The time that patients in the study group spent in the target range was significantly less than in the pooled analysis. The incidence of stroke in the study group (2.0% per year, 0.7% to 4. 4%) was comparable to that of patients receiving warfarin in pooled studies (1.4%, 0.8% to 2.3%). Per year the incidence of major (1.7% v 1.6%) and minor (5.4% v 9.2%) bleeding complications was also similar. CONCLUSION: Rates of stroke and major haemorrhage after anticoagulation in clinical practice were comparable to those obtained from pooled data from randomised controlled studies for patients with atrial fibrillation at high risk of stroke. PMID- 10797032 TI - Establishing a standard definition for child overweight and obesity worldwide: international survey. AB - OBJECTIVE: To develop an internationally acceptable definition of child overweight and obesity, specifying the measurement, the reference population, and the age and sex specific cut off points. DESIGN: International survey of six large nationally representative cross sectional growth studies. SETTING: Brazil, Great Britain, Hong Kong, the Netherlands, Singapore, and the United States. SUBJECTS: 97 876 males and 94 851 females from birth to 25 years of age. MAIN OUTCOME MEASURE: Body mass index (weight/height(2)). RESULTS: For each of the surveys, centile curves were drawn that at age 18 years passed through the widely used cut off points of 25 and 30 kg/m(2) for adult overweight and obesity. The resulting curves were averaged to provide age and sex specific cut off points from 2-18 years. CONCLUSIONS: The proposed cut off points, which are less arbitrary and more internationally based than current alternatives, should help to provide internationally comparable prevalence rates of overweight and obesity in children. PMID- 10797033 TI - Extent of regretted sexual intercourse among young teenagers in Scotland: a cross sectional survey. PMID- 10797035 TI - Human hibernation PMID- 10797034 TI - Burns after photodynamic therapy. PMID- 10797036 TI - Patients' unvoiced agendas in general practice consultations: qualitative study. AB - OBJECTIVE: To investigate patients' agendas before consultation and to assess which aspects of agendas are voiced in the consultation and the effects of unvoiced agendas on outcomes. DESIGN: Qualitative study. SETTING: 20 general practices in south east England and the West Midlands. PARTICIPANTS: 35 patients consulting 20 general practitioners in appointment and emergency surgeries. RESULTS: Patients' agendas are complex and multifarious. Only four of 35 patients voiced all their agendas in consultation. Agenda items most commonly voiced were symptoms and requests for diagnoses and prescriptions. The most common unvoiced agenda items were: worries about possible diagnosis and what the future holds; patients' ideas about what is wrong; side effects; not wanting a prescription; and information relating to social context. Agenda items that were not raised in the consultation often led to specific problem outcomes (for example, major misunderstandings), unwanted prescriptions, non-use of prescriptions, and non adherence to treatment. In all of the 14 consultations with problem outcomes at least one of the problems was related to an unvoiced agenda item. CONCLUSION: Patients have many needs and when these are not voiced they can not be addressed. Some of the poor outcomes in the case studies were related to unvoiced agenda items. This suggests that when patients and their needs are more fully articulated in the consultation better health care may be effected. Steps should be taken in both daily clinical practice and research to encourage the voicing of patients' agendas. PMID- 10797037 TI - Risk of suicide among users of calcium channel blockers: population based, nested case-control study. PMID- 10797039 TI - Online appointment booking to rapid access chest pain clinic PMID- 10797038 TI - Multicentre randomised control trial comparing real time teledermatology with conventional outpatient dermatological care: societal cost-benefit analysis. AB - OBJECTIVES: Comparison of real time teledermatology with outpatient dermatology in terms of clinical outcomes, cost-benefits, and patient reattendance. DESIGN: Randomised controlled trial with a minimum follow up of three months. SETTING: Four health centres (two urban, two rural) and two regional hospitals. SUBJECTS: 204 general practice patients requiring referral to dermatology services; 102 were randomised to teledermatology consultation and 102 to traditional outpatient consultation. MAIN OUTCOME MEASURES: Reported clinical outcome of initial consultation, primary care and outpatient reattendance data, and cost-benefit analysis of both methods of delivering care. RESULTS: No major differences were found in the reported clinical outcomes of teledermatology and conventional dermatology. Of patients randomised to teledermatology, 55 (54%) were managed within primary care and 47 (46%) required at least one hospital appointment. Of patients randomised to the conventional hospital outpatient consultation, 46 (45%) required at least one further hospital appointment, 15 (15%) required general practice review, and 40 (39%) no follow up visits. Clinical records showed that 42 (41%) patients seen by teledermatology attended subsequent hospital appointments compared with 41 (40%) patients seen conventionally. The net societal cost of the initial consultation was pound132.10 per patient for teledermatology and pound48.73 for conventional consultation. Sensitivity analysis revealed that if each health centre had allocated one morning session a week to teledermatology and the average round trip to hospital had been 78 km instead of 26 km, the costs of the two methods of care would have been equal. CONCLUSIONS: Real time teledermatology was clinically feasible but not cost effective compared with conventional dermatological outpatient care. However, if the equipment were purchased at current prices and the travelling distances greater, teledermatology would be a cost effective alternative to conventional care. PMID- 10797040 TI - Science, medicine, and the future. Genetic epidemiology. PMID- 10797041 TI - Congenital neonatal thyrotoxicosis and previous maternal radioiodine therapy. PMID- 10797043 TI - The health benefits from collecting for christian Aid PMID- 10797042 TI - ABC of arterial and vascular disease. Secondary prevention of peripheral vascular disease. PMID- 10797044 TI - Brain stem death: managing care when accepted medical guidelines and religious beliefs are in conflict. Consideration and compromise are possible. PMID- 10797046 TI - Email submissions from outside the united kingdom PMID- 10797045 TI - The function of the discussion section in academic medical writing. PMID- 10797047 TI - Cause of conditions PMID- 10797048 TI - Fallout from the Shipman case. Deaths should be investigated more plainly and effectively. PMID- 10797049 TI - Study on environmental hazards is flawed. PMID- 10797051 TI - Resuscitation decisions are often not documented early enough. PMID- 10797050 TI - Has the sun protection factor had its day? Information on sunscreens should warn against excessive sun exposure. PMID- 10797052 TI - The ethics of unlinked anonymous testing. Surveys provide essential information. PMID- 10797053 TI - Post-traumatic stress disorder in doctors involved in the Omagh bombing. PMID- 10797054 TI - Accidents at home are no more likely in deprived areas. PMID- 10797055 TI - Incidence of hospital admission does not equal incidence of disease. Conclusions drawn from data are incorrect. PMID- 10797056 TI - Social networks are important in preventing dependency in old age. PMID- 10797057 TI - Statins for stroke should be considered in biologically fit people over 75. PMID- 10797058 TI - Language, fairness, and the MRCGP examination. Political correctness going too far. PMID- 10797059 TI - Obituaries PMID- 10797060 TI - BMA responds to proposals on the GMC PMID- 10797061 TI - Neurology and medicine PMID- 10797062 TI - Policy and health: implications for development in asia PMID- 10797063 TI - The psychokiller strikes again PMID- 10797064 TI - Exploring the genome PMID- 10797065 TI - The sins of expertness and a proposal for redemption PMID- 10797067 TI - Experts: off with their heads PMID- 10797066 TI - A medical error PMID- 10797069 TI - Anticoagulation for stroke prophylaxis is effective in practice PMID- 10797068 TI - Birth remains risky for infants in kathmandu PMID- 10797070 TI - Defining an international standard for overweight and obesity in childhood PMID- 10797072 TI - Teledermatology costs more than hospital consultation PMID- 10797071 TI - Patients often have unvoiced agendas in consultations PMID- 10797073 TI - Young teenagers often regret early sexual intercourse PMID- 10797074 TI - Quinolone-induced QT interval prolongation: a not-so-unexpected class effect. PMID- 10797075 TI - Journal of antimicrobial chemotherapy online: a personal guide. PMID- 10797076 TI - Nature of transposon-mediated high-level gentamicin resistance in Enterococcus faecalis isolated in the United Kingdom. AB - Forty-two high-level gentamicin-resistant (MIC > 1000 mg/L) strains of Enterococcus faecalis, isolated from diverse geographical locations throughout the UK between 1993 and 1995, were studied to identify the nature of the high level gentamicin-resistant determinants and the possibility of these determinants being associated with a transposon. High-level gentamicin resistance was attributed to the synthesis of the bifunctional (AAC6'-APH2") aminoglycoside modifying enzyme. The aac6'-aph2" gene, which was present on a 70 kb plasmid in all 42 isolates, could be transferred by conjugation in association with the 70 kb plasmid in 39 of the isolates studied. In three E. faecalis isolates, however, the high-level gentamicin resistance was transferable independent of the 70 kb plasmid, suggesting the presence of a conjugative transposon. Long-PCR studies showed that all 42 clinical isolates harboured a transposon similar to Tn5281, originally identified in E. faecalis strain HH22 isolated in the USA. Restriction endonuclease and Southern hybridization analysis of the UK transposon showed that it is closely related to the high-level gentamicin resistance-conferring transposon Tn5281. However, the UK transposon lacks the HaeIII site identified in Tn5281. Pulsed-field gel electrophoresis analysis identified seven different patterns. Further studies with nine restriction endonucleases showed that the aac6'-aph2" gene was associated with nine different plasmid types in E. faecalis. PMID- 10797077 TI - Detection of tetQ and ermF antibiotic resistance genes in Prevotella and Porphyromonas isolates from clinical specimens and resident microbiota of humans. AB - Gram-negative anaerobes belonging to the genera Fusobacterium, Prevotella and Porphyromonas were investigated for the presence of tetQ and ermF, which have been shown to be spread by conjugal elements. One hundred isolates from either sites of infection or various body sites in healthy subjects were studied. PCR was used to detect tetQ, and DNA-DNA hybridization studies on EcoRI chromosomal digests were undertaken to detect the presence of tetQ and ermF. Antibiotic sensitivity assays were performed on selected isolates to detect tetracycline, erythromycin and penicillin resistance. Twenty Fusobacterium isolates lacked tetQ, and were tetracycline sensitive. Twenty per cent of Prevotella spp. isolates both from clinical specimens and from healthy subjects were found to possess tetQ. Of 20 Porphyromonas isolates tested, one (Porphyromonas levii) from a case of bacterial vaginosis was shown to possess tetQ in the chromosome. The presence of tetQ was always associated with tetracycline resistance. Four isolates of Prevotella melaninogenica and one isolate of Prevotella were ermF positive, although expression of erythromycin resistance was not consistently associated with detection of this gene. Antibiotic resistance phenotypes of Prevotella isolates were shown to be related to specific chromosomal restriction patterns by hybridization studies: tetracycline resistance and tetracycline/erythromycin resistance are conferred by Bacteroides tetracycline resistant ERL elements, whereas the tetracycline/penicillin resistance phenotype could be due to spread of elements identified in Prevotella only. Tetracycline/erythromycin-resistant and tetracycline/erythromycin/penicillin resistant P. melaninogenica isolates were found in this study. It appeared that the presence of tetQ and ermF in Bacteroides and Prevotella contributed to the persistence of antibiotic resistance isolates within the host and to potential spread to other organisms through conjugal elements. PMID- 10797078 TI - Intracellular targets of moxifloxacin: a comparison with other fluoroquinolones. AB - The in vitro activity of the novel 8-methoxyquinolone, moxifloxacin, against Streptococcus pneumoniae was evaluated, and the intracellular targets of this agent were studied. Analysis of mutant strains selected with moxifloxacin demonstrated that first-step mutants bore amino acid substitutions at position 81 in the GyrA subunit of DNA gyrase. This suggests that, unlike older fluoroquinolone agents such as ciprofloxacin and levofloxacin, but similar to other C-8 substituted quinolones like sparfloxacin and gatifloxacin, moxifloxacin targets the GyrA subunit of DNA gyrase as an initial lethal event. Such a mechanism results in high activity against increasingly common S. pneumoniae strains bearing substitutions in DNA topoisomerase IV. Moxifloxacin was active with an MIC of Phe/Tyr substitution in ParC. The moxifloxacin MIC for strains with mutations in the structural genes for both DNA gyrase subunit GyrA and DNA topoisomerase IV subunit ParC did not exceed 2 mg/L, a level within clinically achievable serum concentrations for this agent. We also found that moxifloxacin is a poor substrate for active efflux in S. pneumoniae. Therefore, the high activity of moxifloxacin against S. pneumoniae appears to be a result of both enhanced activity against DNA gyrase and topoisomerase IV, and reduced efflux from the bacterial cell. PMID- 10797079 TI - Use of digoxigenin-labelled ampicillin in the identification of penicillin binding proteins in Helicobacter pylori. AB - Amoxycillin is used in current therapeutic regimens to treat the infection caused by the human gastric pathogen, Helicobacter pylori. The penicillin-binding proteins (PBPs) are the primary targets for the beta-lactam antibiotics, such as amoxycillin, and are involved in the terminal stages of peptidoglycan synthesis. They also play active roles in the determination and maintenance of cellular morphology. It was believed that an organism with a complex morphology, such as H. pylori, would have more than the three PBPs previously suggested. Using digoxigenin-labelled ampicillin (DIG-ampicillin), we report the identification of eight PBPs in H. pylori with masses of 72, 62, 54, 50, 44, 33.5, 30.5 and 28 kDa. A smaller (21 kDa) ninth band was also detected, which may represent another PBP. However, the relatively small size of this apparent PBP raises questions as to whether this is a true PBP. In an attempt to identify the PBPs to which amoxycillin preferentially binds, amoxycillin was used in competition assays with DIG-ampicillin. It appeared that amoxycillin inhibited the binding of DIG ampicillin to only the 72 kDa PBP. The experimental data were also compared with the seven putative PBPs identified in the two published H. pylori genomes, most of which correlate with the experimental data. To investigate further the properties of these PBPs, the seven putative PBP genes identified in the H. pylori genomes were examined. The derived amino acid sequences of the putative PBPs were examined for the three characteristic motifs found in all conventional PBPs, SXXK, SXN and KTG. We were able to determine that all of the putative PBPs had at least one of these motifs, but none possessed all three motifs with the characteristics of conventional PBPs. These findings suggest that the PBPs of H. pylori are unique. PMID- 10797080 TI - Epidemiological studies of large resistance plasmids in Haemophilus. AB - The distribution of large conjugative Haemophilus influenzae plasmids in the nasopharyngeal haemophili of a group of people and in a large collection of 541 H. influenzae type b (Hib) isolates was studied. A newly developed PCR-based assay was used to detect the plasmids. The target sequences were chosen from sequence analysis of part of p1056, a large multiresistance plasmid isolated from a clinical Hib isolate, 1056. Fifty-nine per cent of people were found to carry beta-lactamase-positive (beta-lac(+)), ampicillin-resistant (ampR) haemophili with detectable plasmid sequences. Of these, 83% were in Haemophilus parainfluenzae and 17% were in H. influenzae. In the collection of 541 Hib, antibiotic resistance [beta-lac(+)ampR, beta-lac(+)ampR plus tetracycline resistance (tetR) or tetR] was highly correlated with large plasmids. It was found that 2.3% of the isolates contained large cryptic plasmids (i.e. these isolates were susceptible to antibiotics). The distribution of plasmids between invasive and carried Hib did not differ significantly (25 of 245 and 23 of 276, respectively). Isolates with large plasmids occur at high frequency in the nasopharynx of the normal human population and consist of two populations in Hib, one associated with specific antibiotic resistance traits and the other cryptic. These plasmids do not appear to influence the invasiveness of Hib. PMID- 10797081 TI - High prevalence of erythromycin-resistant, clindamycin/miocamycin-susceptible (M phenotype) Streptococcus pyogenes: results of a Spanish multicentre study in 1998. Spanish Group for the Study of Infection in the Primary Health Care Setting. AB - Using the standard agar dilution method we studied the prevalence of susceptibility to 14-, 15- and 16-membered ring macrolides and clindamycin in Streptococcus pyogenes isolated in 1998 from 21 laboratories in Spain. The number of strains admitted to the study was proportional to the numbers of inhabitants in each geographical area. We also determined the susceptibility phenotypes and the genetic basis for the antibiotic resistance. A total of 486 unduplicated isolates of S. pyogenes were used. Throat swab samples provided 359 (73.9%) isolates, and the remaining 127 isolates were from other sources. One hundred and fourteen (23.5%) isolates were resistant to erythromycin, a 14-membered ring macrolide, and azithromycin, a 15-membered macrolide, whereas only 1% of isolates were resistant to miocamycin, a 16-membered macrolide and 0.8% were resistant to clindamycin. Of the 114 erythromycin-resistant strains, 109 (95.6%) were susceptible to clindamycin and miocamycin. Since induction with erythromycin did not modify susceptibility to the latter antibiotics, these 109 strains were considered to have the M phenotype. Twenty strains with the M phenotype, one per laboratory, were assayed by PCR and showed the presence of the mef gene, which is responsible for antibiotic resistance by an efflux system. Among comparable studies covering entire countries, ours demonstrates one of the highest rates of S. pyogenes erythromycin resistance and clindamycin and miocamycin susceptibility in the world. Strains with the M phenotype account for the great majority of these isolates. PMID- 10797083 TI - In vitro anti-HIV-1 virucidal activity of tyrosine-conjugated tri- and dihydroxy bile salt derivatives. AB - The cellular toxicity and anti-human immunodeficiency virus type 1 (HIV-1) virucidal activity of four synthesized tyrosine-conjugated bile salt derivatives with high surfactant activities, namely di-iodo-deoxycholyltyrosine (DIDCT), di iodo-chenodeoxycholyltyrosine (DICDCT), di-iodo-cholylglycyltyrosine (DICGT) and deoxycholyltyrosine (DCT), were evaluated and compared with either sodium deoxycholate or nonoxynol-9. DIDCT, DICDCT and DCT but not DICGT showed virucidal activity against three different laboratory-adapted strains of HIV-1 (RF, IIIB and MN). All the bile salt derivatives tested excluding DICGT were virucidal at a concentration as low as 10 ng/mL. DCT had the highest anti-HIV-1 virucidal potency, suggesting that monopeptide 7alpha,12alpha dihydroxy bile salt derivatives have the most potent antiviral activity. Complexing of iodine to the bile salt derivative (as in DICGT) decreases virucidal potency. PMID- 10797082 TI - Survey of Klebsiella pneumoniae producing extended-spectrum beta-lactamases at a Portuguese hospital: TEM-10 as the endemic enzyme. AB - One hundred and thirty-eight isolates of Klebsiella pneumoniae showing resistance to ceftazidime were isolated from different wards of the Hospital de Santa Maria, Lisbon. The genomic DNA of the isolates was analysed by pulsed-field gel electrophoresis (PFGE) and two patterns were predominant. In all isolates the presence of a single large plasmid of about 50 kb suggested that propagation of the outbreak prominently involved plasmid spread. The deduced amino acid sequence indicated the presence of a TEM-10 beta-lactamase. This extended-spectrum beta lactamase was present among K. pneumoniae isolates, was widely disseminated in different wards and remained persistent as a result of an outbreak involving the dissemination of both the multi-resistance plasmids harbouring the bla gene and the isolates themselves. PMID- 10797084 TI - Streptococcus pneumoniae in the USA: in vitro susceptibility and pharmacodynamic analysis. AB - Ninety-two laboratories in the USA submitted isolates of Streptococcus pneumoniae to a single laboratory for susceptibility testing. Overall, 64% of 4489 isolates were susceptible to penicillin, 24% were intermediate and 13% were resistant to penicillin, although susceptibilities varied depending on geographical region. Macrolide/azalide resistance varied from 4 to 30%, with some regions having macrolide/azalide resistance higher than penicillin resistance. Children 12 years of age were significantly more likely to be infected with a penicillin-resistant pneumococcus than were adolescents or adults. Isolates from the respiratory tract were more likely to be penicillin resistant and >50% of pneumococci from the ear were resistant to penicillin. Almost 25% of penicillin-susceptible isolates had cefaclor MICs 2.0 mg/L and 15% of penicillin-susceptible isolates had loracarbef MICs 2.0 mg/L. These isolates would be erroneously reported as susceptible using NCCLS guidelines, and this finding may explain the lack of clinical response in patients treated with these antibiotics. The predicted plasma concentrations of all cephalosporins tested exceeded the geometric mean MIC for at least 40% of the dosing interval for penicillin-susceptible S. pneumoniae; for penicillin intermediate S. pneumoniae, only cefprozil (56%), cefuroxime (64%) and cefpodoxime (63%) reached >40% of time above the geometric mean MIC in the dosing interval. None of the cephalosporins evaluated achieved a substantial time above the geometric mean MIC during its dosing interval for fully penicillin-resistant S. pneumoniae. PMID- 10797085 TI - Synergic activity, for anaerobes, of trovafloxacin with clindamycin or metronidazole: chequerboard and time-kill methods. AB - Chequerboard titrations were used to test the activity of trovafloxacin, alone and in combination with clindamycin or metronidazole, against 156 Gram-positive or Gram-negative anaerobes, including 47 Bacteroides fragilis group, 36 Prevotella spp., 26 fusobacteria, 21 peptostreptococci and 26 clostridia. MIC50/MIC90 values (mg/L) of each drug alone against all 156 strains were: trovafloxacin, 0.5/1; clindamycin, 0.25/2; metronidazole, 1/2. Synergy (FIC indices 0. 5-2.0); no antagonism (FIC indices >4.0) was seen. In addition, synergy was tested by time-kill methodology for each of the above combinations against 12 Gram-positive or Gram-negative strains. Results indicated that synergy (defined as a >/= 2 log(10) decrease in cfu/mL at 48 h compared with the more active drug alone) was found between trovafloxacin at or below the MIC and both clindamycin and metronidazole at or below the MIC in one strain each of Bacteroides fragilis, Bacteroides thetaiotaomicron, Prevotella intermedia, Fusobacterium varium, Peptostreptococcus asaccharolyticus and Clostridium bifermentans. Synergy between trovafloxacin (/= 64 mg/L, whereas using the colorimetric method all MICs were >/=16 mg/L. The MIC values of 5-flucytosine were also higher using the reference method (8-16 mg/L for 32% of isolates) compared with the colorimetric method. The percentage of agreement between the methods, using a difference of two dilutions, was 70.7% for itraconazole, 73.2% for amphotericin B, 80% for fluconazole, 88% for 5 flucytosine and 95% for ketoconazole. Overall, we conclude that for fluconazole and 5-flucytosine, in a low but not insignificant number of isolates, results with the two methods are discordant, some isolates being found sensitive with the colorimetric test, but resistant with the reference method. PMID- 10797088 TI - In vitro anti-cryptosporidial activity of cationic peptides alone and in combination with inhibitors of ion transport systems. AB - The anti-cryptosporidial activity of four cationic peptides alone and in combination with five ion transport system (ITS) inhibitors was investigated for six clinical isolates of Cryptosporidium parvum recovered from stools of AIDS patients. The susceptibility tests were performed by inoculating the protozoa on to cell monolayers and determining the parasite count after 48 h incubation at 37 degrees C. The culture medium was supplemented with serial dilutions of cecropin P1, magainin II, indolicidin and ranalexin alone or in combination with amiloride and its analogues. No agent was able to inhibit parasite growth completely. The peptides had some inhibitory effect on parasite growth: cecropin P1, magainin II, indolicidin and ranalexin at a concentration of 50 microM produced 30.6, 33.2, 38.5 and 42.1% reductions, respectively, in schizont count. Conversely, the ITS inhibitors were scarcely effective. Positive interaction was demonstrated when the peptides were tested in combination with ITS inhibitors. PMID- 10797089 TI - Antibiotic resistance in respiratory tract isolates of Haemophilus influenzae and Moraxella catarrhalis collected from across Canada in 1997-1998. AB - Between September 1997 and November 1998 respiratory tract isolates of Haemophilus influenzae (n = 1352) and Moraxella catarrhalis (n = 428) were collected by 18 Canadian medical centres. beta-Lactamase was produced by 24.0 and 94.2% of H. influenzae and M. catarrhalis isolates, respectively. Resistance rates for H. influenzae were highest for ampicillin (24.0%), trimethoprim/sulphamethoxazole (13. 7%), loracarbef (6.1%) and cefaclor (4.2%), and 0.05), indicating a 1:1 stoichiometric relationship. This interpretation must be treated cautiously, because all subjects displayed a greater decrease in TAN compared with the increase in IMP content, and the TAN + IMP + inosine + hypoxanthine content was lower (P < 0.05) immediately after exercise compared with during rest. During the first 5 min of recovery, the increase in TAN was not correlated with the decrease in IMP (r = -0.18, P > 0.05). In all subjects, the magnitude of TAN increase was higher than the magnitude of IMP decrease over this recovery period. In contrast, the increase in TAN was correlated with the decrease in IMP throughout the second 5 min of recovery (r = -0.80, P < 0.05), and it was a 1:1 stoichiometric relationship (slope = -1.12). These data indicate that a small proportion of the TAN pool was temporarily lost from the muscle purine stores during sprinting but was rapidly recovered after exercise. PMID- 10797107 TI - Effects of the RBC membrane and increased perfusate viscosity on hypoxic pulmonary vasoconstriction. AB - Red blood cells (RBCs) augment hypoxic pulmonary vasoconstriction (HPV) in part by scavenging of nitric oxide (NO) by Hb (Deem S, Swenson ER, Alberts MK, Hedges RG, and Bishop MJ, Am J Respir Crit Care Med 157: 1181-1186, 1998). We studied the contribution of the RBC compartmentalization of Hb to augmentation of HPV and scavenging of NO in isolated perfused rabbit lungs. Lungs were initially perfused with buffer; HPV was provoked by a 5-min challenge with hypoxic gas (inspired O(2) fraction 0.05). Expired NO was measured continuously. Addition of free Hb to the perfusate (0.25 mg/ml) resulted in augmentation of HPV and a fall in expired NO that were similar in magnitude to those associated with a hematocrit of 30% (intracellular Hb of 100 mg/ml). Addition of dextran resulted in a blunting of HPV after free Hb but no change in expired NO. Blunting of HPV by dextran was not prevented by NO synthase inhibition with N(omega)-nitro-L-arginine and/or cyclooxygenase inhibition. RBC ghosts had a mild inhibitory effect on HPV but caused a small reduction in expired NO. In conclusion, the RBC membrane provides a barrier to NO scavenging and augmentation of HPV by Hb. Increased perfusate viscosity inhibits HPV by an undetermined mechanism. PMID- 10797108 TI - Effect of different carbohydrate drinks on whole body carbohydrate storage after exhaustive exercise. AB - Seven untrained male subjects participated in a double-blind, crossover study conducted to determine the efficacy of different carbohydrate drinks in promoting carbohydrate storage in the whole body and skeletal muscle during recovery from exhaustive exercise. The postabsorptive subjects first completed an exercise protocol designed to deplete muscle fibers of glycogen, then consumed 330 ml of one of three carbohydrate drinks (18.5% glucose polymer, 18.5% sucrose, or 12% sucrose; wt/vol) and also received a primed constant infusion of [1-(13)C]glucose for 2 h. Nonoxidative glucose disposal (3.51 +/- 0.28, 18.5% glucose polymer; 2.96 +/- 0.32, 18.5% sucrose; 2.97 +/- 0.16, 12% sucrose; all mmol. kg(-1). h( 1)) and storage of muscle glycogen (5.31 +/- 1.11, 18.5% glucose polymer; 4.07 +/ 1.05, 18.5% sucrose; 3.45 +/- 0.85, 12% sucrose; all mmol. kg wet wt(-1). h(-1); P < 0.05) were greater after consumption of the glucose polymer drink than after either sucrose drink. The results suggest that the consumption of a glucose polymer drink (containing 61 g carbohydrate) promotes a more rapid storage of carbohydrate in the whole body, skeletal muscle in particular, than an isoenergetic sucrose drink. PMID- 10797109 TI - A new method for measurement of blood pressure, heart rate, and activity in the mouse by radiotelemetry. AB - A simple and reliable means for accurate, chronic measurement of pulsatile blood pressure (BP) from conscious, freely moving laboratory mice was developed and validated. The newly developed device consists of a small (1.9 ml, 3.4 g), fully implantable radiotelemetry transmitter. Initial frequency response tests showed an adequate dynamic response; the average -3-dB point found in five transmitters was 145 +/- 14 (SD) Hz. BP, heart rate, and locomotor activity were recorded from 16 chronically (30-150 days) implanted mice. Mean arterial and pulse pressure, checked at regular intervals, ranged from 90-140 mmHg and from 30-50 mmHg, respectively, throughout the study. Transmitter BP measurements were validated against a Millar 1.4-Fr. transducer-tipped catheter. The mean error of the transmitters for diastolic pressures was +1.1 +/- 6.9 mmHg (n = 7). The error for systolic pressures was, on average, 2.7 +/- 3.9 mmHg larger. This new device accurately monitors BP, heart rate, and locomotor activity in conscious, untethered, freely moving mice living in their home cages for periods of at least 150 days. PMID- 10797110 TI - Middle cerebral artery blood velocity during a valsalva maneuver in the standing position. AB - Occasionally, lifting of a heavy weight leads to dizziness and even to fainting, suggesting that, especially in the standing position, expiratory straining compromises cerebral perfusion. In 10 subjects, the middle cerebral artery mean blood velocity (V(mean)) was evaluated during a Valsalva maneuver (mouth pressure 40 mmHg for 15 s) both in the supine and in the standing position. During standing, cardiac output decreased by 16 +/- 4 (SE) % (P < 0.05), and at the level of the brain mean arterial pressure (MAP) decreased from 89 +/- 2 to 78 +/- 3 mmHg (P < 0.05), as did V(mean) from 73 +/- 4 to 62 +/- 5 cm/s (P < 0.05). In both postures, the Valsalva maneuver increased central venous pressure by approximately 40 mmHg with a nadir in MAP and cardiac output that was most pronounced during standing (MAP: 65 +/- 6 vs. 87 +/- 3 mmHg; cardiac output: 37 +/- 3 vs. 57 +/- 4% of the resting value; P < 0.05). Also, V(mean) was lowest during the standing Valsalva maneuver (39 +/- 5 vs. 47 +/- 4 cm/s; P < 0.05). In healthy individuals, orthostasis induces an approximately 15% reduction in middle cerebral artery V(mean) that is exaggerated by a Valsalva maneuver performed with 40-mmHg mouth pressure to approximately 50% of supine rest. PMID- 10797111 TI - Fractal nature of regional ventilation distribution. AB - High-resolution measurements of pulmonary perfusion reveal substantial spatial heterogeneity that is fractally distributed. This observation led to the hypothesis that the vascular tree is the principal determinant of regional blood flow. Recent studies using aerosol deposition show similar ventilation heterogeneity that is closely correlated with perfusion. We hypothesize that ventilation has fractal characteristics similar to blood flow. We measured regional ventilation and perfusion with aerosolized and injected fluorescent microspheres in six anesthetized, mechanically ventilated pigs in both prone and supine postures. Adjacent regions were clustered into progressively larger groups. Coefficients of variation were calculated for each cluster size to determine fractal dimensions. At the smallest size lung piece, local ventilation and perfusion are highly correlated, with no significant difference between ventilation and perfusion heterogeneity. On average, the fractal dimension of ventilation is 1.16 in the prone posture and 1. 09 in the supine posture. Ventilation has fractal properties similar to perfusion. Efficient gas exchange is preserved, despite ventilation and perfusion heterogeneity, through close correlation. One potential explanation is the similar geometry of bronchial and vascular structures. PMID- 10797112 TI - Aging, physical conditioning, and exercise-induced changes in hemostatic factors and reaction products. AB - The influence of age on training-induced changes in resting and stimulated hemostatic potential was studied in three age categories (Cat I-III; 20-30 yr, 35 45 yr, and 50-60 yr, respectively) of sedentary men before and after 12 wk of training. Coagulation, fibrinolytic activity, and activation markers (reflecting fibrin formation and degradation) were determined. Physical conditioning resulted in a more pronounced increase in von Willebrand factor (vWF) and factor VIII clotting activity (FVIII:c) in Cat I and II and a more pronounced shortening of the activated partial thromboplastin time in all categories at maximal exertion and during recovery. Enhanced increases in tissue-type plasminogen activator (t PA) antigen and activity and single-chain (sc) urokinase-type plasminogen activator (u-PA) at maximal exercise and 5 min of recovery were observed in all age groups after training. The effects on FVIII:c, vWF, and scu-PA were most pronounced in the youngest age group (Cat I). Increases in the marker of thrombin generation were highest in Cat III; no effect was seen on thrombin-antithrombin complex, plasmin-antiplasmin complex, and D-dimer in any of the age groups. We concluded that training enhances both coagulation and fibrinolytic potential during strenuous exercise. The effect on FVIII/vWF and t-PA/u-PA is most pronounced in younger individuals, whereas thrombin formation is most pronounced in older individuals. PMID- 10797113 TI - Combined aerobic and resistance exercise training improves functional capacity and strength in CHF. AB - This study examined the effect of a novel circuit weight training (CWT) program on cardiorespiratory fitness, muscular strength, and body composition in 13 patients with chronic heart failure (CHF), using a prospective randomized crossover protocol. Peak exercise oxygen uptake (VO(2 peak)) increased after the 8-wk CWT program (19. 5 +/- 1.2 vs. 22.0 +/- 1.5 ml. kg(-1). min(-1), P < 0.01), as did exercise test duration (15.2 +/- 0.9 vs. 18.0 +/- 1.1 min, P < 0. 001). Submaximal exercise heart rate was lower after training at 60 and 80 W (121 +/- 3 vs. 134 +/- 5 beats/min, P < 0.01) as was rate pressure product, whereas ventilatory threshold increased, from 52 +/- 3 to 58 +/- 3% of VO(2 peak) (P < 0.05). CWT also increased maximal isotonic voluntary contractile strength for seven different muscle groups, from 392 to 462 kg (P = 0.001). CWT, an exercise prescription specifically targeting peripheral abnormalities in CHF, improves functional capacity and muscular strength in these patients. PMID- 10797114 TI - No association between the angiotensin-converting enzyme ID polymorphism and elite endurance athlete status. AB - Several studies have reported that the insertion (I) allele of the angiotensin converting enzyme (ACE) I/deletion (D) polymorphism is associated with enhanced responsiveness to endurance training and is more common in endurance athletes than in sedentary controls. We tested the latter hypothesis in a cohort of 192 male endurance athletes with maximal oxygen uptake >/=75 ml. kg(-1). min(-1) and 189 sedentary male controls. The ACE ID polymorphism in intron 16 was typed with the three-primer polymerase chain reaction method. Both the genotype (P = 0.214) and allele (P = 0.095) frequencies were similar in the athletes and the controls. Further analyses in the athletes revealed no excess of the I allele among the athletes within the highest quartile (> 80 ml. kg(-1). min(-1)) or decile (>83 ml. kg(-1). min(-1)) of maximal oxygen uptake. These data from the GENATHLETE cohort do not support the hypothesis that the ACE ID polymorphism is associated with a higher cardiorespiratory endurance performance level. PMID- 10797115 TI - Effect of carbohydrate ingestion on ammonia metabolism during exercise in humans. AB - The present study was undertaken to examine the effect of carbohydrate ingestion on plasma and muscle ammonia (NH(3) denotes ammonia and ammonium) accumulation during prolonged exercise. Eleven trained men exercised for 2 h at 65% peak pulmonary oxygen consumption while ingesting either 250 ml of an 8% carbohydrate electrolyte solution every 15 min (CHO) or an equal volume of a sweet placebo. Blood glucose and plasma insulin levels during exercise were higher in CHO, but plasma hypoxanthine was lower after 120 min (1.7 +/- 0.3 vs. 2.6 +/- 0.1 micromol/l; P < 0. 05). Plasma NH(3) levels were similar at rest and after 30 min of exercise in both trials but were lower after 60, 90, and 120 min of exercise in CHO (62 +/- 9 vs. 76 +/- 9 micromol/l; P < 0.05). Muscle NH(3) levels were similar at rest and after 30 min of exercise but were lower after 120 min of exercise in CHO (1.51 +/- 0.21 vs. 2.07 +/- 0.23 mmol/kg dry muscle; P < 0.05; n = 5). These data are best explained by carbohydrate ingestion reducing muscle NH(3) production from amino acid degradation, although a small reduction in net AMP catabolism within the contracting muscle may also make a minor contribution to the lower tissue NH(3) levels. PMID- 10797116 TI - Regional differences in serotonergic input to canine parasternal intercostal motoneurons. AB - There is a mediolateral gradient in activation of the parasternal intercostal (PI) muscle during inspiration. In the present study, we tested the hypotheses that serotonergic [5-hydroxytryptamine (5-HT)] input from descending central drive and/or intrinsic size-related properties of PI motoneurons leads to the differential activation of PI muscles. In dogs, PI motoneurons innervating the medial and lateral regions of the PI muscles at the T(3)-T(5) interspaces were retrogradely labeled by intramuscular injection of cholera toxin B subunit. After a 10-day survival period, PI motoneurons and 5-HT terminals were visualized by using immunohistochemistry and confocal imaging. There were no differences in motoneuron morphology among motoneurons innervating the medial and lateral regions of the PI muscle. However, the number of 5-HT terminals and the 5-HT terminal density (normalized for surface area) were greater in motoneurons innervating the medial region of the PI muscle compared with the lateral region. These results suggest that differences in distribution of 5-HT input may contribute to regional differences in PI muscle activation during inspiration and that differences in PI motoneuron recruitment do not relate to size. PMID- 10797117 TI - Effects of exercise intensity on the sweating response to a sustained static exercise. AB - To investigate how the sweating response to a sustained handgrip exercise depends on changes in the exercise intensity, the sweating response to exercise was measured in eight healthy male subjects. Each subject lay in the supine position in a climatic chamber (35 degrees C and 50% relative humidity) for approximately 60 min. This exposure caused sudomotor activation by increasing skin temperature without a marked change in internal temperature. After this period, each subject performed isometric handgrip exercise [15, 30, 45, and 60% maximal voluntary contraction (MVC)] for 60 s. Although esophageal and mean skin temperatures did not change with a rise in exercise intensity and were similar at all exercise intensities, the sweating rate (SR) on the forearm increased significantly (P < 0.05) from baseline (0.094 +/- 0.021 mg. cm(-2). min(-1) at 30% MVC, 0.102 +/- 0.022 mg. cm(-2). min(-1) at 45% MVC, 0.059 +/- 0.009 mg. cm(-2). min(-1) at 60% MVC) in parallel with exercise intensity above exercise intensity at 30% MVC (0.121 +/- 0.023 mg. cm(-2). min(-1) at 30% MVC, 0.242 +/- 0.051 mg. cm(-2). min( 1) at 45% MVC, 0.290 +/- 0.056 mg. cm(-2). min(-1) at 60% MVC). Above 45% MVC, SR on the palm increased significantly from baseline (P < 0.05). Although SR on the forearm and palm tended to increase with a rise in exercise intensity, there was a difference in the time courses of SR between sites. SR on the palm showed a plateau after abrupt increase, whereas SR on the forearm increased progressively during exercise. These results suggest that the increase in SR with the increase in sustained handgrip exercise intensity is due to nonthermal factors and that the magnitude of these factors during the exercise may be responsible for the magnitude of SR. PMID- 10797118 TI - Simple contrivance "clamps" end-tidal PCO(2) and PO(2) despite rapid changes in ventilation. AB - The device described in this study uses functionally variable dead space to keep effective alveolar ventilation constant. It is capable of maintaining end-tidal PCO(2) and PO(2) within +/-1 Torr of the set value in the face of increases in breathing above the baseline level. The set level of end-tidal PCO(2) or PO(2) can be independently varied by altering the concentration in fresh gas flow. The device comprises a tee at the mouthpiece, with one inlet providing a limited supply of fresh gas flow and the other providing reinspired alveolar gas when ventilation exceeds fresh gas flow. Because the device does not depend on measurement and correction of end-tidal or arterial gas levels, the response of the device is essentially instantaneous, avoiding the instability of negative feedback systems having significant delay. This contrivance provides a simple means of holding arterial blood gases constant in the face of spontaneous changes in breathing (above a minimum alveolar ventilation), which is useful in respiratory experiments, as well as in functional brain imaging where blood gas changes can confound interpretation by influencing cerebral blood flow. PMID- 10797119 TI - Effect of contractile activity on protein turnover in skeletal muscle mitochondrial subfractions. AB - To determine the role of intramitochondrial protein synthesis (PS) and degradation (PD) in contractile activity-induced mitochondrial biogenesis, we evaluated rates of [(35)S]methionine incorporation into protein in isolated rat muscle subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria. Rates of PS ranged from 47 to 125% greater (P < 0.05) in IMF compared with SS mitochondria. Intense, acute in situ contractile activity (10 Hz, 5 min) of fast-twitch gastrocnemius muscle resulted in a 50% decrease in PS (P < 0.05) in SS but not IMF mitochondria. Recovery, or continued contractile activity (55 min), reestablished PS in SS mitochondria. In contrast, PS was not affected in either SS or IMF mitochondria after prolonged (60-min) contractile activity in the presence or absence of a recovery period. PD was not influenced by 5 min of contractile activity in the presence or absence of recovery but was reduced after 60 min of contractions followed by recovery. Chronic stimulation (10 Hz, 3 h/day, 14 days) increased muscle cytochrome-c oxidase activity by 2.2-fold but reduced PS in IMF mitochondria by 29% (P < 0.05; n = 4). PS in SS mitochondria and PD in both subfractions were not changed by chronic stimulation. Thus acute contractile activity exerts differential effects on protein turnover in IMF and SS mitochondria, and it appears that intramitochondrial PS does not limit the extent of chronic contractile activity-induced mitochondrial biogenesis. PMID- 10797120 TI - Central CO-heme oxygenase pathway raises body temperature by a prostaglandin independent way. AB - Recently, the carbon monoxide (CO)-heme oxygenase pathway has been shown to play an important role in fever generation by acting on the central nervous system, but the mechanisms involved have not been assessed. Thus the present study was designed to determine whether prostagandins participate in the rise in body temperature (T(b)) observed after induction of the CO-heme oxygenase pathway in the central nervous system. Intracerebroventricular (ICV) injection of heme lysinate (152 nmol/4 microl), which is known to induce the CO-heme oxygenase pathway, caused an increase in T(b) [thermal index (TI) = 5.3 +/- 0.5 degrees C. h], which was attenuated by ICV administration of the heme oxygenase inhibitor ZnDPBG (200 nmol/4 microl; TI = 2.5 +/- 1.7 degrees C. h; P < 0.05). No change in T(b) was observed after intraperitoneal injection of the cyclooxygenase inhibitor indomethacin (5 mg/kg), whereas indomethacin at the same dose attenuated the fever induced by ICV administration of lipopolysaccharide (LPS) (10 ng/2 microl) (vehicle/LPS: TI = 4.5 +/- 0.5 degrees C. h; indomethacin/LPS: TI = 1.7 +/- 1.0 degrees C. h; P < 0.05). Interestingly, indomethacin did not affect the rise in T(b) induced by heme-lysinate (152 nmol/4 microl) ICV injection (vehicle/heme: TI = 4.5 +/- 1.4 degrees C. h; indomethacin/heme: TI = 4.2 +/- 1.0 degrees C. h). Finally, PGE(2) (200 ng/2 microl) injected ICV evoked a rise in T(b) that lasted 1.5 h. The heme oxygenase inhibitor ZnDPBG (200 nmol/4 microl) failed to alter PGE(2)-induced fever. Taken together, these results indicate that the central CO heme oxygenase pathway increases T(b) independently of prostaglandins. PMID- 10797121 TI - Embryonic quail eye development in microgravity. AB - The US-Russian joint quail embryo project was designed to study the effects of microgravity on development of Japanese quail embryos incubated aboard Mir. For this part of the project, eyes from embryonic days 14 and 16 (E14 and E16) flight embryos were compared with eyes from several groups of ground-based control embryos. Measurements were recorded for eye weights; eye, corneal, and scleral ring diameters; and numbers of bones in scleral ossicle rings. Transparency of E16 corneas was documented, and immunohistochemical staining was performed to observe corneal innervation. In addition, corneal ultrastructure was observed at the electron microscopic level. Except for corneal diameter of E16 flight embryos, compared with that of one of the sets of controls, results reported here indicate that eye development occurred normally in microgravity. Fixation by cracking the shell and placing the egg in paraformaldehyde solution did not adequately preserve corneal nerves or cellular ultrastructure. PMID- 10797122 TI - Ciliary neurotrophic factor prevents unweighting-induced functional changes in rat soleus muscle. AB - The purpose of the present work was to see whether changes in rat soleus characteristics due to 3 wk of hindlimb suspension could be modified by ciliary neurotrophic factor (CNTF) treatment. Throughout the tail suspension period, the cytokine was delivered by means of an osmotic pump (flow rate 16 microg. kg(-1). h(-1)) implanted under the hindlimb skin. In contrast to extensor digitorum longus, CNTF treatment was able to reduce unweighting-induced atrophy in the soleus. Twitch and 146 mM potassium (K) tensions, measured in small bundles of unloaded soleus, decreased by 48 and 40%, respectively. Moreover, the time to peak tension and the time constant of relaxation of the twitch were 48 and 54% faster, respectively, in unloaded soleus than in normal muscle. On the contrary, twitch and 146 mM K contracture generated in CNTF-treated unloaded and normal soleus were not different. CNTF receptor-alpha mRNA expression increased in extensor digitorum longus and soleus unloaded nontreated muscles but was similar in CNTF-treated unloaded muscles. The present results demonstrate that exogenously provided CNTF could prevent functional changes occurring in soleus innervated muscle subject to unweighting. PMID- 10797123 TI - Muscle glycogen resynthesis during recovery from cycle exercise: no effect of additional protein ingestion. AB - In the present study, we have investigated the effect of carbohydrate and protein hydrolysate ingestion on muscle glycogen resynthesis during 4 h of recovery from intense cycle exercise. Five volunteers were studied during recovery while they ingested, immediately after exercise, a 600-ml bolus and then every 15 min a 150 ml bolus containing 1) 1.67 g. kg body wt(-1). l(-1) of sucrose and 0.5 g. kg body wt(-1). l(-1) of a whey protein hydrolysate (CHO/protein), 2) 1.67 g. kg body wt(-1). l(-1) of sucrose (CHO), and 3) water. CHO/protein and CHO ingestion caused an increased arterial glucose concentration compared with water ingestion during 4 h of recovery. With CHO ingestion, glucose concentration was 1-1.5 mmol/l higher during the first hour of recovery compared with CHO/protein ingestion. Leg glucose uptake was initially 0.7 mmol/min with water ingestion and decreased gradually with no measurable glucose uptake observed at 3 h of recovery. Leg glucose uptake was rather constant at 0.9 mmol/min with CHO/protein and CHO ingestion, and insulin levels were stable at 70, 45, and 5 mU/l for CHO/protein, CHO, and water ingestion, respectively. Glycogen resynthesis rates were 52 +/- 7, 48 +/- 5, and 18 +/- 6 for the first 1.5 h of recovery and decreased to 30 +/- 6, 36 +/- 3, and 8 +/- 6 mmol. kg dry muscle(-1). h(-1) between 1.5 and 4 h for CHO/protein, CHO, and water ingestion, respectively. No differences could be observed between CHO/protein and CHO ingestion ingestion. It is concluded that coingestion of carbohydrate and protein, compared with ingestion of carbohydrate alone, did not increase leg glucose uptake or glycogen resynthesis rate further when carbohydrate was ingested in sufficient amounts every 15 min to induce an optimal rate of glycogen resynthesis. PMID- 10797124 TI - Involvement of cGMP-dependent protein kinase in the relaxation of ovine pulmonary arteries to cGMP and cAMP. AB - Agonist-induced smooth muscle relaxation occurs following an increase in intracellular concentrations of cGMP or cAMP. However, the role of protein kinase G (PKG) and/or protein kinase A (PKA) in cGMP- or cAMP-mediated pulmonary vasodilation is not clearly elucidated. In this study, we examined the relaxation responses of isolated pulmonary arteries of lambs (age = 10 +/- 1 days), preconstricted with endothelin-1, to increasing concentrations of 8-bromo-cGMP (8 BrcGMP) or 8-BrcAMP (cell-permeable analogs), in the presence or absence of Rp-8 beta-phenyl-1,N(2)-etheno-bromoguanosine cyclic monosphordthioate (Rp-8-PET BrcGMPS) or KT-5720, selective inhibitors of PKG and PKA, respectively. When examined for specificity, Rp-8-Br-PET-cGMPS abolished PKG, but not PKA, activity in pulmonary arterial extracts, whereas KT-5720 inhibited PKA activity only. 8 BrcGMP-induced relaxation was inhibited by the PKG inhibitor only, whereas 8 BrcAMP-induced relaxation was inhibited by both inhibitors. A nearly fourfold higher concentration of cAMP than cGMP was required to relax arteries by 50% and to activate PKG by 50%. Our results demonstrate that relaxation of pulmonary arteries is more sensitive to cGMP than cAMP and that PKG plays an important role in both cGMP- and cAMP-mediated relaxation. PMID- 10797125 TI - Estrogen modifies the temperature effects of progesterone. AB - To test the hypothesis that progestin-mediated increases in resting core temperature and the core temperature threshold for sweating onset are counteracted by estrogen, we studied eight women (24 +/- 2 yr) at 27 degrees C rest, during 20 min of passive heating (35 degrees C), and during 40 min of exercise at 35 degrees C. Subjects were tested four times, during the early follicular and midluteal menstrual phases, after 4 wk of combined estradiol norethindrone (progestin) oral contraceptive administration (OC E+P), and after 4 wk of progestin-only oral contraceptive administration (OC P). The order of the OC P and OC E+P were randomized. Baseline esophageal temperature (T(es)) at 27 degrees C was higher (P < 0.05) in the luteal phase (37.08 +/- 0.21 degrees C) and in OC P (37.60 +/- 0.31 degrees C) but not during OC E+P (37.04 +/- 0.23 degrees C) compared with the follicular phase (36.66 +/- 0.21 degrees C). T(es) remained above follicular phase levels throughout passive heating and exercise during OC P, whereas T(es) in the luteal phase was greater than in the follicular phase throughout exercise (P < 0.05). The T(es) threshold for sweating was also greater in the luteal phase (38.02 +/- 0.28 degrees C) and OC P (38.07 +/- 0.17 degrees C) compared with the follicular phase (37.32 +/- 0.11 degrees C) and OC E+P (37.46 +/- 0.18 degrees C). Progestin administration raised the T(es) threshold for sweating during OC P, but this effect was not present when estrogen was administered with progestin, suggesting that estrogen modifies progestin related changes in temperature regulation. These data are also consistent with previous findings that estrogen lowers the thermoregulatory operating point. PMID- 10797127 TI - Ventilation heterogeneity in excised lobes: effect of tidal volume. AB - Although several factors are known to influence nonuniformity of ventilation, including lung mechanical properties (regional structure and compliance), external factors (chest wall, pleural pressure, heart), and ventilatory parameters (tidal and preinspiratory volume, flow rate), their relative contributions are poorly understood. We studied five excised, unperfused, canine right-middle lobes under varied levels of tidal volume (VT), thus eliminating many factors affecting heterogeneity. Multiple-breath washouts of N(2) were analyzed for anatomic dead space volume (VD(anat)), nonuniformity of N(2) washout, and nonuniformity between joined acinar regions vs. that occurring between larger joined regions. Approximately 80% of ventilation heterogeneity was found among joined acinar regions at resting levels of VT, but increasing VT reduced intra-acinar heterogeneity to about 25% of that found at resting levels. Increasing VT had essentially no effect on VD(anat) and heterogeneity among larger joined regions. The results indicate that the magnitude of VT is a major influence on the dominant intra-acinar component of ventilation heterogeneity and that VT effects on VD(anat) are likely due to perfusion and/or influences normally external to the lobar structure. PMID- 10797126 TI - Cardiac output during exercise by the open circuit acetylene washin method: comparison with direct Fick. AB - An open-circuit (OpCirc) acetylene uptake cardiac output (QT) method was modified for use during exercise. Two computational techniques were used. OpCirc1 was based on the integrated uptake vs. end-tidal change in acetylene, and OpCirc2 was based on an iterative finite difference modeling method. Six subjects [28-44 yr, peak oxygen consumption (VO(2)) = 120% predicted] performed cycle ergometry exercise to compare QT using OpCirc and direct Fick methods. An incremental protocol was repeated twice, separated by a 10-min rest, and subsequently subjects exercised at 85-90% of their peak work rate. Coefficient of variation of the OpCirc methods and Fick were highest at rest (OpCirc1, 7%, OpCirc2, 12%, Fick, 10%) but were lower at moderate to high exercise intensities (OpCirc1, 3%, OpCirc2, 3%, Fick, 5%). OpCirc1 and OpCirc2 QT correlated highly with Fick QT (R(2) = 0.90 and 0.89, respectively). There were minimal differences between OpCirc1 and OpCirc2 compared with Fick up to moderate-intensity exercise (<70% peak VO(2)); however, both techniques tended to underestimate Fick at >70% peak VO(2). These differences became significant for OpCirc1 only. Part of the differences between Fick and OpCirc methods at the higher exercise intensities are likely related to inhomogeneities in ventilation and perfusion matching (R(2) = 0.36 for Fick - OpCirc1 vs. alveolar-to-arterial oxygen tension difference). In conclusion, both OpCirc methods provided reproducible, reliable measurements of QT during mild to moderate exercise. However, only OpCirc2 appeared to approximate Fick QT at the higher work intensities. PMID- 10797128 TI - Effects of duration of positive-pressure ventilation on blood-brain barrier function in premature lambs. AB - We have been studying the ontogeny of the blood-brain barrier function in ovine fetuses and lambs. During these studies, we have found that the duration of ventilation also influences blood-brain barrier permeability in premature lambs. Chronically instrumented hysterotomy-delivered surfactant-treated premature lambs were studied at 90% or 137 days of gestation (n = 9). Blood-brain barrier function was quantified with the blood-to-brain transfer constant K(i) to alpha aminoisobutyric acid. Linear regression analysis was used to compare the K(i) values in the brain regions, as the dependent variable, to the duration of ventilation, as the independent variable. There were direct correlations (P < 0.05) between the K(i) values and the duration of ventilation [306 min (mean), 162-474 min (range)] in the cerebral cortex, cerebellum, medulla, caudate nucleus, hippocampus, superior colliculus, inferior colliculus, thalamus, pons, cervical spinal cord, and choroid plexus, but not in the pituitary gland. Ventilatory pressures and rates were established before the onset of the permeability studies. Calculated mean airway pressures [14 cmH(2)O (mean), 7-20 cmH(2)O (range)] from similarly studied premature lambs did not correlate with the duration of positive-pressure ventilation. We conclude that increases in the duration of positive-pressure ventilation predispose premature lambs to increases in regional blood-brain barrier permeability. These alterations in barrier function occur over relatively short time intervals (minutes to hours). In our study, these changes in permeability are most likely not attributable to changes in mean airway pressure. PMID- 10797129 TI - Force production of the genioglossus as a function of muscle length in normal humans. AB - Resting muscle length affects both maximum force production and force maintenance. The strength and force maintenance characteristics of the genioglossus as a function of resting muscle length have not been described. We hypothesized that genioglossus optimum length (L(o)) could be defined in vivo and that the ability of the genioglossus to sustain a given workload would decrease as resting length deviated from L(o). To test this, 11 normal men repeated maximal isometric genioglossus protrusions at different muscle lengths to determine L(o). L(o) was also obtained by using submaximal efforts while simultaneously recording electromyographic activity of the genioglossus, with L(o) defined as the length at which the force-to-genioglossus electromyographic activity ratio was maximum. Both methods provided similar results. Force maintenance was measured at four muscle lengths on separate days. Target efforts representing 60% of each subject's maximum at L(o) and lasting 5 s were performed at 12-s intervals. Time limit of endurance of the genioglossus was defined as the time from trial onset at which 90% of the target could not be maintained for three consecutive efforts. Time limit of endurance was greatest at L(o) and fell to 47.5% at L(o) + 1 cm, 53.8% at L(o) - 1 cm, and 47.4% at L(o) - 1.5 cm. We conclude that L(o) of the genioglossus can be determined in vivo and that force maintenance of the genioglossus is decreased when operating length deviates from L(o). PMID- 10797130 TI - Comparative effects of alpha-receptor stimulation and nitrergic inhibition on bronchovascular tone. AB - Adrenergic agonists are known to influence bronchial blood flow and bronchovascular resistance. Recently, the nitrergic system has also been implicated in the control of bronchovascular tone. In this study, we compared the effects of the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) and the alpha(1)-receptor agonist phenylephrine on bronchovascular resistance in anesthetized sheep (n = 9). Bronchial blood flow, cardiac output, and systemic and pulmonary arterial pressures were continuously monitored. Phenylephrine (1.2-3.4 microg. kg(-1). min(-1)) was infused intravenously to increase mean systemic arterial pressure above 95 Torr for 10 min and then was discontinued. When hemodynamic parameters returned to baseline, nebulized phenylephrine (10 mg) was given over 10 min. When parameters again normalized, L NAME (30 mg/kg) was infused intravenously over 1 min. Intravenous phenylephrine increased systemic vascular resistance by 40% at 10 min with no concurrent increase in bronchovascular resistance, but inhaled phenylephrine increased bronchovascular resistance by 66% at 10 min. By comparison, intravenous L-NAME produced a rapid and sustained fivefold increase in bronchovascular resistance at 10 min. We conclude that, although alpha-agonist stimulation has some influence on bronchovascular resistance in sheep, the nitrergic system has predominant control of bronchovascular tone. PMID- 10797131 TI - Effect of captopril on skeletal muscle angiogenic growth factor responses to exercise. AB - Acute exercise increases vascular endothelial growth factor (VEGF), transforming growth factor-beta(1) (TGF-beta(1)), and basic fibroblast growth factor (bFGF) mRNA levels in skeletal muscle, with the greatest increase in VEGF mRNA. VEGF functions via binding to the VEGF receptors Flk-1 and Flt-1. Captopril, an angiotensin-converting enzyme inhibitor, has been suggested to reduce the microvasculature in resting and exercising skeletal muscle. However, the molecular mechanisms responsible for this reduction have not been investigated. We hypothesized that this might occur via reduced VEGF, TGF-beta(1), bFGF, Flk-1, and Flt-1 gene expression at rest and after exercise. To investigate this, 10-wk old female Wistar rats were placed into four groups (n = 6 each): 1) saline + rest; 2) saline + exercise; 3) 100 mg/kg ip captopril + rest; and 4) 100 mg/kg ip captopril + exercise. Exercise consisted of 1 h of running at 20 m/min on a 10 degrees incline. VEGF, TGF-beta(1), bFGF, Flk-1, and Flt-1 mRNA were analyzed from the left gastrocnemius by quantitative Northern blot. Exercise increased VEGF mRNA 4.8-fold, TGF-beta(1) mRNA 1.6-fold, and Flt-1 mRNA 1.7-fold but did not alter bFGF or Flk-1 mRNA measured 1 h after exercise. Captopril did not affect the rest or exercise levels of VEGF, TGF-beta(1), bFGF, and Flt-1 mRNA. Captopril did reduce Flk-1 mRNA 30-40%, independently of exercise. This is partially consistent with the suggestion that captopril may inhibit capillary growth. PMID- 10797132 TI - Possible mechanisms of muscle cramp from temporal and spatial surface EMG characteristics. AB - In this study, the initiation and development of muscle cramp are investigated. For this, we used a 64-channel surface electromyogram (EMG) to study the triceps surae muscle during both cramp and maximal voluntary contraction (MVC) in four cramp-prone subjects and during cramp only in another four cramp-prone subjects. The results show that cramp presents itself as a contraction of a slowly moving fraction of muscle fibers, indicating that either the spatial arrangement of the motoneurons and muscle fibers is highly related or that cramp spreads at a level close to the muscle. Spectral analyses of the EMG and peak-triggered average potentials show the presence of extremely short potentials during cramp compared with during MVC. These results can also be interpreted in two ways. Either the motoneurons fire with enlarged synchronization during MVC compared with cramp, or smaller units than motor units are active, indicating that cramp is initiated close to or even at the muscle fiber level. Further research is needed to draw final conclusions. PMID- 10797134 TI - Ventilatory and metabolic adaptations to walking and cycling in patients with COPD. AB - To test the hypothesis that in chronic obstructive pulmonary disease (COPD) patients the ventilatory and metabolic requirements during cycling and walking exercise are different, paralleling the level of breathlessness, we studied nine patients with moderate to severe, stable COPD. Each subject underwent two exercise protocols: a 1-min incremental cycle ergometer exercise (C) and a "shuttle" walking test (W). Oxygen uptake (VO(2)), CO(2) output (VCO(2)), minute ventilation (VE), and heart rate (HR) were measured with a portable telemetric system. Venous blood lactates were monitored. Measurements of arterial blood gases and pH were obtained in seven patients. Physiological dead space-tidal volume ratio (VD/VT) was computed. At peak exercise, W vs. C VO(2), VE, and HR values were similar, whereas VCO(2) (848 +/- 69 vs. 1,225 +/- 45 ml/min; P < 0. 001) and lactate (1.5 +/- 0.2 vs. 4.1 +/- 0.2 meq/l; P < 0.001) were lower, DeltaVE/DeltaVCO(2) (35.7 +/- 1.7 vs. 25.9 +/- 1.3; P < 0. 001) and DeltaHR/DeltaVO(2) values (51 +/- 3 vs. 40 +/- 4; P < 0.05) were significantly higher. Analyses of arterial blood gases at peak exercise revealed higher VD/VT and lower arterial partial pressure of oxygen values for W compared with C. In COPD, reduced walking capacity is associated with an excessively high ventilatory demand. Decreased pulmonary gas exchange efficiency and arterial hypoxemia are likely to be responsible for the observed findings. PMID- 10797133 TI - Substrate metabolism during different exercise intensities in endurance-trained women. AB - We have studied eight endurance-trained women at rest and during exercise at 25, 65, and 85% of maximal oxygen uptake. The rate of appearance (R(a)) of free fatty acids (FFA) was determined by infusion of [(2)H(2)]palmitate, and fat oxidation rates were determined by indirect calorimetry. Glucose kinetics were assessed with [6,6-(2)H(2)]glucose. Glucose R(a) increased in relation to exercise intensity. In contrast, whereas FFA R(a) was significantly increased to the same extent in low- and moderate-intensity exercise, during high-intensity exercise, FFA R(a) was reduced compared with the other exercise values. Carbohydrate oxidation increased progressively with exercise intensity, whereas the highest rate of fat oxidation was during exercise at 65% of maximal oxygen uptake. After correction for differences in lean body mass, there were no differences between these results and previously reported data in endurance-trained men studied under the same conditions, except for slight differences in glucose metabolism during low-intensity exercise (Romijn JA, Coyle EF, Sidossis LS, Gastaldelli A, Horowitz JF, Endert E, and Wolfe RR. Am J Physiol Endocrinol Metab 265: E380-E391, 1993). We conclude that the patterns of changes in substrate kinetics during moderate- and high-intensity exercise are similar in trained men and women. PMID- 10797135 TI - Inhibition of allergic late airway responses by inhaled heparin-derived oligosaccharides. AB - Inhaled heparin has been shown to inhibit allergic bronchoconstriction in sheep that develop only acute responses to antigen (acute responders) but was ineffective in sheep that develop both acute and late airway responses (LAR) (dual responders). Because the antiallergic activity of heparin is molecular weight dependent, we hypothesized that heparin-derived oligosaccharides (<2, 500) with potential anti-inflammatory activity may attenuate the LAR in the dual responder sheep. Specific lung resistance was measured in 24 dual-responder sheep before and serially for 8 h after challenge with Ascaris suum antigen for demonstration of early airway response (EAR) and LAR, without and after treatment with inhaled medium-, low-, and ultralow-molecular-weight (ULMW) heparins and "non-anticoagulant" fractions (NAF) of heparin. Airway responsiveness was estimated before and 24 h postantigen as the cumulative provocating dose of carbachol that increased specific lung resistance by 400%. Only ULMW heparins caused a dose-dependent inhibition of antigen-induced EAR and LAR and postantigen airway hyperresponsiveness (AHR), whereas low- and medium-molecular-weight heparins were ineffective. The effects of ULMW heparin and ULMW NAF-heparin were comparable and inhibited the LAR and AHR even when administered "after" the antigen challenge. The ULMW NAF-heparin failed to inhibit the bronchoconstrictor response to histamine, carbachol, and leukotriene D(4), excluding a direct effect on airway smooth muscle. In six sheep, segmental antigen challenge caused a marked increase in bronchoalveolar lavage histamine, which was not prevented by inhaled ULMW NAF-heparin. The results of this study in the dual-responder sheep demonstrate that 1) the antiallergic activity of inhaled "fractionated" heparins is molecular-weight dependent, 2) only ULMW heparins inhibit the antigen-induced EAR and LAR and postantigen AHR, and 3) the antiallergic activity is mediated by nonanticoagulant fractions and resides in the ULMW chains of <2,500. PMID- 10797136 TI - Exercise and bone mineral density in men: a meta-analysis. AB - The purpose of this study was to use the meta-analytic approach to examine the effects of exercise on bone mineral density (BMD) in men. A total of 26 effect sizes (ES) representing 225 subjects from 8 studies met the criteria for inclusion. When BMD sites assessed were specific to the sites loaded during exercise, increases of approximately 2.6% (2.1% in the exercisers and -0.5% in the controls) were found. These results were statistically significant (ES = 0.213, 95% bootstrap confidence interval = 0.007-0.452). Statistically significant ES changes were found for older (>31 yr) but not younger (<31 yr) adults, with differences between groups statistically significant (P = 0.04). Statistically significant changes were also observed at the femur, lumbar, and os calcis sites. The results of this study suggest that site-specific exercise may help improve and maintain BMD at the femur, lumbar, and os calcis sites in older men. However, the biological importance of the small changes observed for most outcomes, quality of studies, and limited data pool prevent us from forming any firm conclusion regarding the use of exercise for maintaining and/or improving BMD in men. Clearly, a need exists for additional studies. PMID- 10797137 TI - Cholinergic dopamine release from the in vitro rabbit carotid body. AB - The aim of this study was to test whether cholinergic mechanisms regulate dopamine (DA) release from the carotid body (CB) and interact with DA D(2) autoreceptors. One hundred forty-two CBs from adult rabbits were infused in vitro in a surviving medium bubbled with O(2) (Bairam A, Marchal F, Cottet-Emard JM, Basson H, Pequignot JM, Hascoet JM, and Lahiri S. J Appl Physiol 80: 20-24, 1996). CB DA content and release were measured after 1 h of exposure to various treatments: control, cholinergic agonist (0.1-50 microM carbachol), full muscarinic antagonist (1 and 10 microM atropine), antagonists of M(1) and M(2) muscarinic receptors (1 and 10 microM pirenzepine and 10 microM AFDX-116, respectively), and the DA D(2) receptor antagonist domperidone (1 microM), alone and with carbachol (1 microM). Compared with control, the release of DA was significantly increased by carbachol (1-50 microM), AFDX-116, and domperidone and decreased by atropine (10 microM) and pirenzepine (10 microM). The effects of domperidone and carbachol were not significantly different but were clearly additive. It is concluded that, in the rabbit CB, M(1) and M(2) muscarinic receptor subtypes may be involved in the control of DA release, in addition to the DA D(2) autoreceptors. PMID- 10797138 TI - Impairment of Ca(2+) release in single Xenopus muscle fibers fatigued at varied extracellular PO(2). AB - We tested the hypothesis that the mechanisms involved in the more rapid onset of fatigue when O(2) availability is reduced in contracting skeletal muscle are similar to those when O(2) availability is more sufficient. Two series of experiments were performed in isolated, single skeletal muscle fibers from Xenopus laevis. First, relative force and free cytosolic Ca(2+) concentrations ([Ca(2+)](c)) were measured simultaneously in single fibers (n = 6) stimulated at increasing frequencies (0.25, 0.33, 0.5, and 1 Hz) at an extracellular PO(2) of either 22 or 159 Torr. Muscle fatigue (force = 50% of initial peak tension) occurred significantly sooner (P < 0.05) during the low- (237 +/- 40 s) vs. high PO(2) treatments (280 +/- 38 s). Relative [Ca(2+)](c) was significantly decreased from maximal values at the fatigue time point during both the high- (72 +/- 4%) and low-PO(2) conditions (78 +/- 4%), but no significant difference was observed between the treatments. In the second series of experiments, using the same stimulation regime as the first, fibers (n = 6) exposed to 5 mM caffeine immediately after fatigue demonstrated an immediate but incomplete relative force recovery during both the low- (89 +/- 4%) and high-PO(2) treatments (82 +/- 3%), with no significant difference between treatments. Additionally, there was no significant difference in relative [Ca(2+)](c) between the high- (100 +/- 12% of prefatigue values) and low-PO(2) treatments (108 +/- 12%) on application of caffeine. These results suggest that in isolated, single skeletal muscle fibers, the earlier onset of fatigue that occurred during the low-extracellular PO(2) condition was modulated through similar pathways as the fatigue process during the high and involved a decrease in relative peak [Ca(2+)](c). PMID- 10797139 TI - Cardiac troponin T alterations in myocardium and serum of rats after stressful, prolonged intense exercise. AB - The goal of this study was to determine whether the stress of forced exercise would result in injury to the myocardium. Male rats with 8% of body weight attached to the tail were forced to swim 3.5 h (3.5S), forced to swim 5 h (5S), or pretrained for 8 days and then forced to swim 5 h (T5S). Rats were killed immediately after they swam (0 h PS) and at 3 h (3 h PS), 24 h (24 h PS), and 48 h after they swam (48 h PS). Tissue homogenates of the left ventricle were analyzed by Western blot analysis for cardiac troponin T (cTnT). Serum cTnT was quantified by immunoassay. Results indicated that, in the 3.5S, 5S, and T5S groups, serum cTnT was significantly (P < 0.01) increased at 0 and 3 h PS. The 5S group demonstrated a greater increase in serum cTnT than the 3.5S group (P < 0.01) and the T5S group (P < 0.01) at 0 h PS. Western blot analysis indicated significant decreases (P < 0. 01) in myocardial cTnT in the 5S group only at 0 h PS (P < 0.01) and 3 h PS (P < 0.05). Histological evidence of localized myocyte damage demonstrated by interstitial inflammatory infiltrates consisting of neutrophils, lymphocytes, and histiocytes, as well as vesicular nuclei-enlarged chromatin patterns, was observed in left ventricle specimens from the 5S group at 24 and 48 h PS. Our findings demonstrate that stressful, forced exercise induces alterations in myocardial cTnT and that training before exercise attenuates the exercise-induced heart damage. PMID- 10797140 TI - Reflex control of the cutaneous circulation during passive body core heating in humans. AB - The impact of body core heating on the interaction between the cutaneous and central circulation during blood pressure challenges was examined in eight adults. Subjects were exposed to -10 to -90 mmHg lower body negative pressure (LBNP) in thermoneutral conditions and -10 to -60 mmHg LBNP during heat stress. We measured forearm vascular conductance (FVC; ml. min(-1). 100 ml(-1). mmHg(-1)) by plethysmography; cutaneous vascular conductance (CVC) by laser-Doppler techniques; and central venous pressure, arterial blood pressure, and cardiac output by impedance cardiography. Heat stress increased FVC from 5.7 +/- 0.9 to 18.8 +/- 1.3 conductance units (CU) and CVC from 0.21 +/- 0.07 to 1.02 +/- 0.20 CU. The FVC-CVP relationship was linear over the entire range of LBNP and was shifted upward during heat stress with a slope increase from 0. 46 +/- 0.10 to 1.57 +/- 0.3 CU/mmHg CVP (P < 0.05). Resting CVP was lower during heat stress (6.3 +/- 0.6 vs. 7.7 +/- 0.6 mmHg; P < 0. 05) but fell to similar levels during LBNP as in normothermic conditions. Data analysis indicates an increased capacity, but not sensitivity, of peripheral baroreflex responses during heat stress. Laser-Doppler techniques detected thermoregulatory responses in the skin, but no significant change in CVC occurred during mild-to-moderate LBNP. Interestingly, very high levels of LBNP produced cutaneous vasodilation in some subjects. PMID- 10797141 TI - Glucose infusion attenuates endogenous glucose production and enhances glucose use of horses during exercise. AB - We examined the effects of increased glucose availability on glucose kinetics and substrate utilization in horses during exercise. Six conditioned horses ran on a treadmill for 90 min at 34 +/- 1% of maximum oxygen uptake. In one trial [glucose (Glu)], glucose was infused at a mean rate of 34.9 +/- 1.1 micromol. kg(-1). min( 1), whereas in the other trial [control (Con)] an equivalent volume of isotonic saline was infused. Plasma glucose increased during exercise in Glu (90 min: 8.3 +/- 1.7 mM) but was largely unchanged in Con (90 min: 5.1 +/- 0.4 mM). In Con, hepatic glucose production (HGP) increased during exercise, reaching a peak of 38.6 +/- 2.7 micromol. kg(-1). min(-1) after 90 min. Glucose infusion partially suppressed (P < 0.05) the rise in HGP (peak value 25.8 +/- 3.3 micromol. kg(-1). min(-1)). In Con, glucose rate of disappearance (R(d)) rose to a peak of 40.4 +/- 2.9 micromol. kg(-1). min(-1) after 90 min; in Glu, augmented glucose utilization was reflected by values for glucose R(d) that were twofold higher (P < 0.001) than in Con between 30 and 90 min. Total carbohydrate oxidation was higher (P < 0.05) in Glu (187.5 +/- 8.5 micromol. kg(-1). min(-1)) than in Con (159.2 +/- 7.3 micromol. kg(-1).min(-1)), but muscle glycogen utilization was similar between trials. We conclude that an increase in glucose availability in horses during low intensity exercise 1) only partially suppresses HGP, 2) attenuates the decrease in carbohydrate oxidation during such exercise, but 3) does not affect muscle glycogen utilization. PMID- 10797142 TI - Epinephrine inhibits exogenous glucose utilization in exercising horses. AB - This study examined the effects of preexercise glucose administration, with and without epinephrine infusion, on carbohydrate metabolism in horses during exercise. Six horses completed 60 min of treadmill exercise at 55 +/- 1% maximum O(2) uptake 1) 1 h after oral administration of glucose (2 g/kg; G trial); 2) 1 h after oral glucose and with an intravenous infusion of epinephrine (0.2 micromol. kg(-1). min(-1); GE trial) during exercise, and 3) 1 h after water only (F trial). Glucose administration (G and GE) caused hyperinsulinemia and hyperglycemia ( approximately 8 mM). In GE, plasma epinephrine concentrations were three- to fourfold higher than in the other trials. Compared with F, the glucose rate of appearance was approximately 50% and approximately 33% higher in G and GE, respectively, during exercise. The glucose rate of disappearance was approximately 100% higher in G than in F, but epinephrine infusion completely inhibited the increase in glucose uptake associated with glucose administration. Muscle glycogen utilization was higher in GE [349 +/- 44 mmol/kg dry muscle (dm)] than in F (218 +/- 28 mmol/kg dm) and G (201 +/- 35 mmol/kg dm). We conclude that 1) preexercise glucose augments utilization of plasma glucose in horses during moderate-intensity exercise but does not alter muscle glycogen usage and 2) increased circulating epinephrine inhibits the increase in glucose rate of disappearance associated with preexercise glucose administration and increases reliance on muscle glycogen for energy transduction. PMID- 10797143 TI - Exercise modulates antioxidant enzyme gene expression in rat myocardium and liver. AB - Our previous studies have shown that exercise caused changes in the tissue activities of the antioxidant enzymes glutathione peroxidase, superoxide dismutase, and catalase in spontaneously hypertensive (SH) and Wistar-Kyoto (WKY) rats. To determine whether the changes observed were due to changes in mRNA levels of the enzymes, levels of tissue mRNA were determined by quantitative RNase protection assay. Comparisons of tissue enzyme activities and mRNA levels in sedentary and exercised animals showed that, in some cases (e.g., glutathione peroxidase in SH and WKY myocardium), parallel changes in enzyme activity and mRNA levels occurred, whereas in other cases (e.g., catalase in SH and WKY liver), nonparallel changes were found. Exercise of hypertensive rats altered antioxidant enzyme mRNA levels to those seen in normotensive animals in some, but not all, cases. The results suggest that transcriptional control over changes in exercise-related antioxidant enzyme activities is operative in some cases, although in other cases posttranscriptional regulatory mechanisms may exist. PMID- 10797144 TI - L-Arginine increases nitric oxide production in isolated lungs of chronically hypoxic newborn pigs. AB - Previously, our laboratory found that pulmonary hypertension developed and lung nitric oxide (NO) production was reduced when piglets were exposed to chronic hypoxia (Fike CD, Kaplowitz MR, Thomas CJ, and Nelin LD. Am J Physiol Lung Cell Mol Physiol 274: L517-L526, 1998). The purposes of this study were to determine whether L-arginine addition augments NO production and to evaluate whether L arginine uptake is impaired in isolated lungs of chronically hypoxic newborn piglets. Studies were performed by using 1- to 3-day-old piglets raised in room air (control) or 10% O(2) (chronic hypoxia) for 10-12 days. Lung NO production was assessed in isolated lungs from both groups by measuring the perfusate accumulation of nitrites and nitrates (collectively termed NO(-)(x)) before and after addition of L-arginine (10(-2) M) to the perfusate. The rate of perfusate NO(-)(x) accumulation increased by 220% (from 0.8 +/- 0.4 to 2.5 +/- 0.5 nmol/min, P < 0.05) after L-arginine addition to chronic hypoxic lungs but remained unchanged (3.2 +/- 0. 8 before vs. 3.3 +/- 0.4 nmol/min after L arginine) in control lungs. In the second series of studies, L-arginine uptake was evaluated by measuring the perfusate concentration of L-[(3)H]arginine at fixed time intervals. The perfusate concentration of L-[(3)H]arginine at each time point was less (P < 0.05) in control than in chronic hypoxic lungs. Thus L arginine uptake was impaired and may underlie in part the reduction in lung NO production that occurs when piglets are exposed to 10-12 days of chronic hypoxia. Moreover, these findings in isolated lungs lead to the possibility that L arginine supplementation might increase in vivo lung NO production in piglets with chronic hypoxia-induced pulmonary hypertension. PMID- 10797145 TI - Age causes a redistribution of joint torques and powers during gait. AB - At self-selected walking speeds, elderly compared with young adults generate decreased joint torques and powers in the lower extremity. These differences may be actual gait-limiting factors and neuromuscular adaptations with age or simply a consciously selected motor pattern to produce a slower gait. The purpose of the study was to compare joint torques and powers of young and elderly adults walking at the same speed. Twelve elderly and fourteen young adults (ages 69 and 21 yr) walked at 1.48 m/s over a force platform while being videotaped. Hip, knee, and ankle torques and powers were calculated from the reaction force and kinematic data. A support torque was calculated as the sum of the three joint torques. Extensor angular impulse during stance and positive work at each joint were derived from the torques and powers. Step length was 4% shorter and cadence was 4% higher in elderly adults (both P < 0.05) compared with young adults. Support angular impulse was nearly identical between groups, but elderly adults had 58% greater angular impulse and 279% more work at the hip, 50% less angular impulse and 39% less work at the knee, and 23% less angular impulse and 29% less work at the ankle compared with young adults (t-test, all P < 0.05). Age caused a redistribution of joint torques and powers, with the elderly using their hip extensors more and their knee extensors and ankle plantar flexors less than young adults when walking at the same speed. Along with a reduction in motor and sensory functions, the natural history of aging causes a shift in the locus of function in motor performance. PMID- 10797146 TI - Kinetics of oxygen uptake at the onset of exercise near or above peak oxygen uptake. AB - We tested the hypothesis that kinetics of O(2) uptake (VO(2)) measured in the transition to exercise near or above peak VO(2) (VO(2 peak)) would be slower than those for subventilatory threshold exercise. Eight healthy young men exercised at approximately 57, approximately 96, and approximately 125% VO(2 peak). Data were fit by a two- or three-component exponential model and with a semilogarithmic transformation that tested the difference between required VO(2) and measured VO(2). With the exponential model, phase 2 kinetics appeared to be faster at 125% VO(2 peak) [time constant (tau(2)) = 16.3 +/- 8.8 (SE) s] than at 57% VO(2 peak) (tau(2) = 29. 4 +/- 4.0 s) but were not different from that at 96% VO(2 peak) exercise (tau(2) = 22.1 +/- 2.1 s). VO(2) at the completion of phase 2 was 77 and 80% VO(2 peak) in tests predicted to require 96 and 125% VO(2 peak). When VO(2) kinetics were calculated with the semilogarithmic model, the estimated tau(2) at 96% VO(2 peak) (49.7 +/- 5.1 s) and 125% VO(2 peak) (40.2 +/- 5.1 s) were slower than with the exponential model. These results are consistent with our hypothesis and with a model in which the cardiovascular system is compromised during very heavy exercise. PMID- 10797147 TI - Endocrine markers of semistarvation in healthy lean men in a multistressor environment. AB - We tested the hypothesis that key endocrine responses to semistarvation would be attenuated by changing only the food intake in a multistressor environment that also included sustained workload, inadequate sleep, and thermal strain. Serum hormones were compared within and between two groups of healthy young male volunteers participating in the 8-wk US Army Ranger course, with four repeated cycles of restricted energy intakes and refeeding: group 1 (n = 49) and group 2 (n = 48); energy deficits averaged 1,200 and 1,000 kcal/day, respectively. After 8 wk, most of group 1 achieved a minimum body fat, serum 3,5,3'-triiodothyronine (T(3)) was below normal (78 +/- 20 ng/dl), testosterone (T) approached castrate levels (4.5 +/- 3.9 nmol/l), insulin-like growth factor I (IGF-I) declined by one half (75 +/- 25 microg/l), and cholesterol rose from 158 +/- 31 to 217 +/- 39 mg/dl. Bioavailable T(3) and T were further reduced by increases in their specific binding proteins in response to declining insulin. Refeeding, even with continuation of the other stressors, produced prompt recovery of T(3), T, and IGF I. Higher energy intakes in group 2 attenuated the subclinical hypothyroidism and hypercholesterolemia, whereas consistent luteinizing hormone suppression indicated centrally mediated threshold effects on gonadal hormone suppression. We conclude that low T, T(3), and IGF-I remained reliable markers of acute energy deficits in the presence of other stressors; elevated cholesterol and cortisol provided information about chronic status, corresponding to diminishing body fat stores. PMID- 10797149 TI - Ventilatory responses to specific CNS hypoxia in sleeping dogs. AB - Our study was concerned with the effect of brain hypoxia on cardiorespiratory control in the sleeping dog. Eleven unanesthetized dogs were studied; seven were prepared for vascular isolation and extracorporeal perfusion of the carotid body to assess the effects of systemic [and, therefore, central nervous system (CNS)] hypoxia (arterial PO(2) = 52, 45, and 38 Torr) in the presence of a normocapnic, normoxic, and normohydric carotid body during non-rapid eye movement sleep. A lack of ventilatory response to systemic boluses of sodium cyanide during carotid body perfusion demonstrated isolation of the perfused carotid body and lack of other significant peripheral chemosensitivity. Four additional dogs were carotid body denervated and exposed to whole body hypoxia for comparison. In the sleeping dog with an intact and perfused carotid body exposed to specific CNS hypoxia, we found the following. 1) CNS hypoxia for 5-25 min resulted in modest but significant hyperventilation and hypocapnia (minute ventilation increased 29 +/- 7% at arterial PO(2) = 38 Torr); carotid body-denervated dogs showed no ventilatory response to hypoxia. 2) The hyperventilation was caused by increased breathing frequency. 3) The hyperventilatory response developed rapidly (<30 s). 4) Most dogs maintained hyperventilation for up to 25 min of hypoxic exposure. 5) There were no significant changes in blood pressure or heart rate. We conclude that specific CNS hypoxia, in the presence of an intact carotid body maintained normoxic and normocapnic, does not depress and usually stimulates breathing during non-rapid eye movement sleep. The rapidity of the response suggests a chemoreflex meditated by hypoxia-sensitive respiratory-related neurons in the CNS. PMID- 10797148 TI - Effect of age on sleep onset-related changes in respiratory pump and upper airway muscle function. AB - In normal young men, there is an abrupt fall in ventilation (VE), a rise in upper airway resistance (UAR), and falls in the activities of the diaphragm (Di), intercostals (IC), genioglossus (GG), and tensor palatini (TP) at sleep onset. On waking, there is an abrupt increase in VE and fall in UAR and an increase in the activities of Di, IC, GG, and TP. The aim of this study was to determine whether these changes are age dependent. Nine men aged 20 to 25 yr were compared with nine men aged 42 to 67 yr. Airflow, UAR, Di, and IC surface electromyograms (EMGs) and the intramuscular EMGs of GG and TP were recorded. It was found that the falls in IC, GG, and TP at the transition from alpha to theta electroencephalogram (EEG) activity were significantly greater in the older than in the younger men (P < 0.05) and that the fall in Di was also greater, although this was only marginally significant (P = 0.15). The rise in GG at theta-to-alpha transitions was also greater in the older than in the younger men, and there was a trend for TP to be higher. PMID- 10797150 TI - Light-dark differences in the effects of ambient temperature on gaseous metabolism in newborn rats. AB - Body temperature (T(b)) of rat pups (7-9 days old) raised under a 12:12-h light dark (L-D) regimen (L: 0700-1900, D: 1900-0700) was consistently higher in D than in L by approximately 1.1 degrees C. We tested the hypothesis that the L-D differences in T(b) were accompanied by differences in the set point of thermoregulation. Measurements were performed on rat pups at 7-9 days after birth. O(2) consumption (VO(2)) and CO(2) production (VCO(2)) were measured with an open-flow method during air breathing, as ambient temperature (T(a)) was decreased from 40 to 15 degrees C at the constant rate of 0.5 degrees C/min. At T(a) >/=33 degrees C, VO(2) was not significantly different between L and D, whereas VCO(2) was higher in L, suggesting a greater ventilation. Over the 33 to 15 degrees C range the VO(2) values in D exceeded those in L by approximately 30%. Specifically, the difference was contributed by differences in thermogenesis at T(a) = 30 to 20 degrees C. As T(a) was decreased, the critical temperature at which VO(2) began to rise was lower in L. We conclude that the higher T(b) of rat pups in D is accompanied by a higher set point for thermoregulation and a greater thermogenesis. These results are consistent with the idea that, in newborns, endogenous changes in the set point of thermoregulation contribute to the circadian oscillations of T(b). PMID- 10797151 TI - Respiratory sensation during chest wall restriction and dead space loading in exercising men. AB - We mimicked important mechanical and ventilatory aspects of restrictive lung disorders by employing chest wall strapping (CWS) and dead space loading (DS) in normal subjects to gain mechanistic insights into dyspnea causation and exercise limitation. We hypothesized that thoracic restriction with increased ventilatory stimulation would evoke exertional dyspnea that was similar in nature to that experienced in such disorders. Twelve healthy young men [28 +/- 2 (SE) yr of age] completed pulmonary function tests and maximal cycle exercise tests under four conditions, in randomized order: 1) control, 2) CWS to 60% of vital capacity, 3) added DS of 600 ml, and 4) CWS + DS. Measurements during exercise included cardiorespiratory parameters, esophageal pressure, and Borg scale ratings of dyspnea. Compared with control, CWS significantly reduced the tidal volume response to exercise, increased dyspnea intensity at any given work rate or ventilation, and thus limited exercise performance. DS stimulated ventilation but had minimal effects on dyspnea and exercise performance. Adding DS to CWS further increased dyspnea by 1.7 +/- 0.6 standardized Borg units (P = 0.012) and decreased exercise performance (total work) by 21 +/- 6% (P = 0.003) over CWS alone. Across conditions, increased dyspnea intensity correlated best with decreased resting inspiratory reserve volume (r = -0.63, P < 0.0005). Dyspnea during CWS was described primarily as "inspiratory difficulty" and "unsatisfied inspiration," similar to restrictive disorders. In conclusion, severe dyspnea and exercise intolerance were provoked in healthy normal subjects when tidal volume responses were constrained in the face of increased ventilatory drive during exercise. PMID- 10797152 TI - Forced expiration: a test for airflow obstruction in horses. AB - The purpose of this study was to assess whether our method of inducing forced expiration detects small airway obstruction in horses. Parameters derived from forced expiratory flow-volume (FEFV) curves were compared with lung mechanics data obtained during spontaneous breathing in nine healthy horses, in three after histamine challenge, and in two with chronic obstructive pulmonary disease (COPD) pre- and posttherapy with prednisone. Parameters measured in the healthy horses included forced vital capacity (FVC = 41.6 +/- 5.8 liters; means +/- SD) and forced expiratory flow (FEF) at various percentages of FVC (range of 20.4-29.7 l/s). Histamine challenge induced a dose-dependent decrease in FVC and FEF at low lung volume. After therapy, lung function of the two COPD horses improved to a point where one horse had normal lung mechanics during tidal breathing; however, FEF at 95% of FVC (4.9 l/s) was still decreased. We concluded that FEFV curve analysis allowed the detection of induced or naturally occurring airway obstruction. PMID- 10797153 TI - Cellular oxygen sensing by mitochondria: old questions, new insight. AB - Hypoxia elicits a variety of adaptive responses at the tissue level, at the cellular level, and at the molecular level. A physiological response to hypoxia requires the existence of an O(2) sensor coupled to a signal transduction system, which in turn activates the functional response. Although much has been learned about the signaling systems activated by hypoxia, no consensus exists regarding the nature of the underlying O(2) sensor or whether multiple sensors exist. Among previously considered mechanisms, heme proteins have been suggested to undergo allosteric modification in response to O(2) binding or release at different PO(2) levels. Other studies suggest that ion channels may change conductance as a function of PO(2), allowing them to signal the onset of hypoxia. Still other studies suggest that NADPH oxidase may decrease its generation of reactive O(2) species (ROS) during hypoxia. Recent data suggest that mitochondria may function as O(2) sensors by increasing their generation of ROS during hypoxia. These oxidant signals appear to act as second messengers in the adaptive responses to hypoxia in a variety of cell types. Such observations contribute to a growing awareness that mitochondria do more than just generate ATP, in that they initiate signaling cascades involved in adaptive responses to hypoxia and that they participate in the control of cell death pathways. PMID- 10797154 TI - Hypoxia regulates gene expression of alveolar epithelial transport proteins. AB - Alveolar hypoxia occurs during ascent to high altitude but is also commonly observed in many acute and chronic pulmonary disorders. The alveolar epithelium is directly exposed to decreases in O(2) tension, but a few studies have evaluated the effects of hypoxia on alveolar cell function. The alveolar epithelium consists of two cell types: large, flat, squamous alveolar type I and cuboidal type II (ATII). ATII cells are more numerous and have a number of critical functions, including transporting ions and substrates required for many physiological processes. ATII cells express 1) membrane proteins used for supplying substrates required for cell metabolism and 2) ion transport proteins such as Na(+) channels and Na(+)-K(+)-ATPase, which are involved in the vectorial transport of Na(+) from the alveolar to interstitial spaces and therefore drive the resorption of alveolar fluid. This brief review focuses on gene expression regulation of glucose transporters and Na(+) transport proteins by hypoxia in alveolar epithelial cells. Cells exposed to severe hypoxia (0% or 3% O(2)) for 24 h upregulate the activity and expression of the glucose transporter GLUT-1, resulting in preservation of ATP content. Hypoxia-induced increases in GLUT-1 mRNA levels are due to O(2) deprivation and inhibition of oxidative phosphorylation. This regulation occurs at the transcriptional level through activation of a hypoxia-inducible factor. In contrast, hypoxia downregulates expression and activity of Na(+) channels and Na(+)-K(+)-ATPase in cultured alveolar epithelial cells. Hypoxia induces time- and concentration-dependent decreases of alpha-, beta-, and gamma-subunits of epithelial Na(+) channel mRNA and beta(1)- and alpha(1)-subunits of Na(+)-K(+)-ATPase, effects that are completely reversed after reoxygenation. The mechanisms by which O(2) deprivation regulates gene expression of Na(+) transport proteins are not fully elucidated but likely involve the redox status of the cell. Thus hypoxia regulates gene expression of transport proteins in cultured alveolar epithelial type II cells differently, preserving ATP content. PMID- 10797155 TI - L-type Ca(2+) channel activation regulates induction of c-fos transcription by hypoxia. AB - In the present study we examined the intracellular pathways that link hypoxia to activation of c-fos gene expression. Experiments were performed on rat pheocromocytoma-12 (PC-12) cells. c-fos mRNA and promoter activities were analyzed by RT-PCR and reporter gene assays, respectively. BAPTA, a Ca(2+) chelator, inhibited c-fos mRNA and promoter activation by hypoxia. Nitrendipine, an L-type Ca(2+)-channel blocker, abolished, whereas BAY K 8644, an L-type channel agonist, enhanced c-fos activation by hypoxia. Ca(2+) currents were augmented reversibly by hypoxia, suggesting that Ca(2+) influx mediated by L-type Ca(2+) channels is essential for c-fos activation by hypoxia. We next determined downstream pathways activated by intracellular Ca(2+) concentration. Immunoblot analysis revealed Ca(2+)/calmodulin-dependent kinase II (CaMKII) protein in PC-12 cells and revealed that hypoxia increased the enzyme activity. KN-93, a CaMK inhibitor, blocked CaMKII activation and c-fos promoter stimulation by hypoxia. Ectopic expression of an active mutant of CaMKII (pCaMKII290) stimulated c-fos promoter activity under normoxia. Hypoxia increased phosphorylation of CREB at the serine residue 133 (Ser-133), and KN-93 attenuated this effect. Point mutations at the Ca(2+)/cAMP-responsive cis-element (Ca/CRE) attenuated, whereas point mutations in the serum-responsive cis-element (SRE) abolished transcriptional activation of c-fos by hypoxia. These results demonstrate that c fos activation by hypoxia involves CaMK activation and CREB phosphorylation at Ser-133 and requires Ca/CRE and SRE. These observations demonstrate that Ca(2+) dependent signaling pathways play a crucial role in induction of c-fos gene expression, which may underlie long-term adaptive responses to hypoxia. PMID- 10797156 TI - Development of the ventilatory response to hypoxia in Swiss CD-1 mice. AB - We examined developmental changes in breathing pattern and the ventilatory response to hypoxia (7.4% O(2)) in unanesthetized Swiss CD-1 mice ranging in age from postnatal day 0 to 42 (P(0)-P(42)) using head-out plethysmography. The breathing pattern of P(0) mice was unstable. Apneas were frequent at P(0) (occupying 29 +/- 6% of total time) but rare by P(3) (5 +/- 2% of total time). Tidal volume increased in proportion to body mass ( approximately 10-13 ml/kg), but increases in respiratory frequency (f) (55 +/- 7, 130 +/- 13, and 207 +/- 20 cycles/min for P(0), P(3), and P(42), respectively) were responsible for developmental increases in minute ventilation (690 +/- 90, 1,530 +/- 250, and 2,170 +/- 430 ml. min(-1). kg(-1) for P(0), P(3), and P(42), respectively). Between P(0) and P(3), increases in f were mediated by reductions in apnea and inspiratory and expiratory times; beyond P(3), increases were due to reductions in expiratory time. Mice of all ages showed a biphasic hypoxic ventilatory response, which differed in two respects from the response typical of most mammals. First, the initial hyperpnea, which was greatest in mature animals, decreased developmentally from a maximum, relative to control, of 2.58 +/- 0.29 in P(0) mice to 1. 32 +/- 0.09 in P(42) mice. Second, whereas ventilation typically falls to or below control in most neonatal mammals, ventilation remained elevated relative to control throughout the hypoxic exposure in P(0) (1.73 +/- 0.31), P(3) (1.64 +/- 0.29), and P(9) (1. 34 +/- 0.17) mice but not in P(19) or P(42) mice. PMID- 10797157 TI - Consequences of periodic augmented breaths on tongue muscle activities in hypoxic rats. AB - This study was designed to investigate the influence of hypoxia-evoked augmented breaths (ABs) on respiratory-related tongue protrudor and retractor muscle activities and inspiratory pump muscle output. Genioglossus (GG) and hyoglossus (HG) electromyogram (EMG) activities and respiratory-related tongue movements were compared with peak esophageal pressure (Pes; negative change in pressure during inspiration) and minute Pes (Pes x respiratory frequency = Pes/min) before and after ABs evoked by sustained poikilocapnic, isocapnic, and hypercapnic hypoxia in spontaneously breathing, anesthetized rats. ABs evoked by poikilocapnic and isocapnic hypoxia triggered long-lasting (duration at least 10 respiratory cycles) reductions in GG and HG EMG activities and tongue movements relative to pre-AB levels, but Pes was reduced transiently (duration of <10 respiratory cycles) after ABs. Adding 7% CO(2) to the hypoxic inspirate had no effect on the frequency of evoked ABs, but this prevented long-term declines in tongue muscle activities. Bilateral vagotomy abolished hypoxia-induced ABs and stabilized drive to the tongue muscles during each hypoxic condition. We conclude that, in the rat, hypoxia-evoked ABs 1) elicit long-lasting reductions in protrudor and retractor tongue muscle activities, 2) produce short-term declines in inspiratory pump muscle output, and 3) are mediated by vagal afferents. The more prolonged reductions in pharyngeal airway vs. pump muscle activities may lead to upper airway narrowing or collapse after spontaneous ABs. PMID- 10797158 TI - Methodological and physiological variability within the ventilatory response to hypoxia in humans. AB - Measurement of the acute hypoxic ventilatory response (AHVR) requires careful choice of the hypoxic stimulus. If the stimulus is too brief, the response may be incomplete; if the stimulus is too long, hypoxic ventilatory depression may ensue. The purpose of this study was to compare three different techniques for assessing AHVR, using different hypoxic stimuli, and also to examine the between day variability in AHVR. Ten subjects were studied, each on six different occasions, which were >/=1 wk apart. On each occasion, AHVR was assessed using three different protocols: 1) protocol SW, which uses square waves of hypoxia; 2) protocol IS, which uses incremental steps of hypoxia; and 3) protocol RB, which simulates an isocapnic rebreathing test. Mean values for hypoxic sensitivity were 1.02 +/- 0.48, 1.15 +/- 0.55, and 0.93 +/- 0.60 (SD) l. min(-1). %(-1) for protocols SW, IS, and RB, respectively. These differed significantly (P < 0.01). The coefficients of variation for measurement of AHVR were 20, 23, and 36% for the three protocols, respectively. These were not significantly different. There was a significant physiological variation in AHVR (F (50,100) = 3.9, P < 0. 001), with a coefficient of variation of 26%. We conclude that there was relatively little systematic variation between the three protocols but that AHVR varies physiologically over time. PMID- 10797159 TI - Ethnicity and stroke : beware of the fallacies. PMID- 10797160 TI - There's no place like home : an evaluation of early supported discharge for stroke. AB - BACKGROUND AND PURPOSE: Because stroke management is aimed at facilitating community reintegration, it would be logical that the sooner the patient can be discharged home, the sooner reintegration can commence. The purpose of this study was to determine the effectiveness of prompt discharge combined with home rehabilitation on function, community reintegration, and health-related quality of life during the first 3 months after stroke. METHODS: A randomized trial was carried out involving patients who required rehabilitation services and who had a caregiver at home. When medically ready for discharge, persons with stroke were randomized to either the home intervention group (n=58) or the usual care group (n=56). The home group received a 4-week, tailor-made home program of rehabilitation and nursing services; persons randomized to the usual care group received services provided through a variety of mechanisms, depending on institutional, care provider, and personal preference. The main outcome measure was the Physical Health component of the Measuring Outcomes Study Short-Form-36 (SF-36). Associated outcomes measures included the Timed Up & Go (TUG), Barthel Index (BI), the Older Americans Resource Scale for instrumental activities of daily living (OARS-IADL), Reintegration to Normal Living (RNL), and the SF-36 Mental Health component. RESULTS: The total length of stay for the home group was, on average, 10 days, 6 days shorter than that for the usual care group. There were no differences between the 2 groups on the BI or on the TUG at either 1 or 3 months after stroke; however, there was a significantly beneficial impact of the home intervention on IADL and reintegration (RNL). By 3 months after stroke, the home intervention group showed a significantly higher score on the SF 36 Physical Health component than the usual care group. The total number of services received by the home group was actually lower than that received by the usual care group. CONCLUSIONS: Prompt discharge combined with home rehabilitation appeared to translate motor and functional gains that occur through natural recovery and rehabilitation into a greater degree of higher-level function and satisfaction with community reintegration, and these in turn were translated into a better physical health. PMID- 10797161 TI - Home or hospital for stroke rehabilitation? results of a randomized controlled trial : I: health outcomes at 6 months. AB - BACKGROUND AND PURPOSE: We wished to examine the effectiveness of an early hospital discharge and home-based rehabilitation scheme for patients with acute stroke. METHODS: This was a randomized, controlled trial comparing early hospital discharge and home-based rehabilitation with usual inpatient rehabilitation and follow-up care. The trial was carried out in 2 affiliated teaching hospitals in Adelaide, South Australia. Participants were 86 patients with acute stroke (mean age, 75 years) who were admitted to hospital and required rehabilitation. Forty two patients received early hospital discharge and home-based rehabilitation (median duration, 5 weeks), and 44 patients continued with conventional rehabilitation care after randomization. The primary end point was self-reported general health status (SF-36) at 6 months after randomization. A variety of secondary outcome measures were also assessed. RESULTS: Overall, clinical outcomes for patients did not differ significantly between the groups at 6 months after randomization, but the total duration of hospital stay in the experimental group was significantly reduced (15 versus 30 days; P<0.001). Caregivers among the home-based rehabilitation group had significantly lower mental health SF-36 scores (mean difference, 7 points). CONCLUSIONS: A policy of early hospital discharge and home-based rehabilitation for patients with stroke can reduce the use of hospital rehabilitation beds without compromising clinical patient outcomes. However, there is a potential risk of poorer mental health on the part of caregivers. The choice of this management strategy may therefore depend on convenience and costs but also on further evaluations of the impact of stroke on caregivers. PMID- 10797162 TI - Home or hospital for stroke Rehabilitation? Results of a randomized controlled trial : II: cost minimization analysis at 6 months. AB - BACKGROUND AND PURPOSE: The goal of the present study was to examine the resource and economic implications of an early hospital discharge and home-based rehabilitation scheme for patients with acute stroke. METHODS: A cost minimization analysis in conjunction with a randomized controlled trial was carried out at 2 affiliated teaching hospitals in the southern metropolitan region of Adelaide, South Australia, between 1997 and 1998. Eighty-six hospitalized patients with acute stroke who required rehabilitation were randomized to receive both early hospital discharge and home-based rehabilitation, or conventional in-hospital rehabilitation and community care. Direct and indirect costs related to stroke rehabilitation were calculated, including hospital bed days, home-based intervention program, community services, and personal expenses during the 6 months after randomization. RESULTS: The mean cost per patient was lower for patients randomized to the early hospital discharge and home-based rehabilitation ($8040) compared with those who received conventional care ($10 054). This cost saving was not statistically significant (P=0.14). However, sensitivity analyses indicated that the cost of home-based rehabilitation was consistently lower than that of conventional care except when hospital costs were assumed to be 50% less than those used in the main analysis. Multiple regression analysis demonstrated that the cost of the home-based program was significantly related to a patient's level of disability after adjustment for age, comorbidity, and the presence or absence of a caregiver. CONCLUSIONS: The early hospital discharge and home-based rehabilitation scheme was less costly than conventional hospital care for patients with stroke. Limitation of the provision of such services to patients with mild disability is likely to be most cost effective. PMID- 10797163 TI - Can readmission after stroke be prevented? Results of a randomized clinical study: a postdischarge follow-up service for stroke survivors. AB - BACKGROUND AND PURPOSE: About 50% of stroke survivors are discharged to their homes with lasting disability. Knowledge, however, of the importance of follow-up services that targets these patients is sparse. The purpose of the present study was to evaluate 2 models of follow-up intervention after discharge. The study hypothesis was that intervention could reduce readmission rates and institutionalization and prevent functional decline. We report the results regarding readmission. METHODS: This randomized study included 155 stroke patients with persistent impairment and disability who, after the completion of inpatient rehabilitation, were discharged to their homes. The patients were randomized to 1 of 2 follow-up interventions provided in addition to standard care or to standard aftercare. Fifty-four received follow-up home visits by a physician (INT1-HVP), 53 were provided instructions by a physiotherapist in their home (INT2-PI), and 48 received standard aftercare only (controls). Baseline characteristics for the 3 groups were comparable. Six months after discharge, data were obtained on readmission and institutionalization. RESULTS: The readmission rates within 6 months after discharge were significantly lower in the intervention groups than in the control group (INT1-HVP 26%, INT2-PI 34%, controls 44%; P=0.028). Multivariate analysis of readmission risk showed a significant favorable effect of intervention (INT1-HVP or INT2-PI) in interaction with length of hospital stay (P=0.0332), indicating that the effect of intervention was strongest for patients with a prolonged inpatient rehabilitation. CONCLUSIONS: Readmission is common among disabled stroke survivors. Follow-up intervention after discharge seems to be a way of preventing readmission, especially for patients with long inpatient rehabilitation. PMID- 10797164 TI - Economic evaluation in stroke research : a systematic review. AB - BACKGROUND AND PURPOSE: The purpose of this review is to provide insight into the quality of economic evaluation in the field of cerebrovascular diseases (CVD) on the basis of a systematic analysis. METHODS: A literature search was performed using several sources. Trial-based full economic evaluation studies, were included in this review. The quality of the studies was independently assessed by 2 reviewers using a checklist. RESULTS: Twenty-three articles were found to comply with our inclusion and exclusion criteria. Only a few studies mentioned the perspective of the study, and in these cases it was always the societal perspective. The majority of the studies were cost-minimization and cost effectiveness analyses based on cohort studies. All studies included healthcare costs, and in some instances patient and family costs were considered. Costs were usually measured by tariffs. Clinical end points and mortality were used to measure effects. Cost and effect measurements were based on hospital records. CONCLUSIONS: Only a few full economic evaluations have been undertaken in the domain of CVD. In most of the studies, the technical execution and methodology were limited. PMID- 10797165 TI - A multinational comparison of subarachnoid hemorrhage epidemiology in the WHO MONICA stroke study. AB - BACKGROUND AND PURPOSE: By official, mostly unvalidated statistics, mortality from subarachnoid hemorrhage (SAH) show large variations between countries. Using uniform criteria for case ascertainment and diagnosis, a multinational comparison of attack rates and case fatality rates of SAH has been performed within the framework of the WHO MONICA Project. METHODS: In 25- to 64-year-old men and women, a total of 3368 SAH events were recorded during 35.9 million person-years of observation in 11 populations in Europe and China. Strict MONICA criteria were used for case ascertainment and diagnosis of stroke subtype. Case fatality was based on follow-up at 28 days after onset. RESULTS: Age-adjusted average annual SAH attack rates varied 10-fold among the 11 populations studied, from 2.0 (95% CI 1.6 to 2.4) per 100 000 population per year in China-Beijing to 22.5 (95% CI 20.9 to 24.1) per 100 000 population per year in Finland. No consistent pattern was observed in the sex ratio of attack rates in the different populations. The overall 28-day case fatality rate was 42%, with 2-fold differences in age adjusted rates between populations but little difference between men and women. Case fatality rates were consistently higher in Eastern than in Western Europe. CONCLUSIONS: Using a uniform methodology, the WHO MONICA Project has shown very large variations in attack rates of SAH across 11 populations in Europe and China. The generally accepted view that women have a higher risk of SAH than men does not apply to all populations. Marked differences in outcome of SAH add to the wide gap in the burden of stroke between East and West Europe. PMID- 10797166 TI - Ischemic stroke subtypes : a population-based study of functional outcome, survival, and recurrence. AB - BACKGROUND AND PURPOSE: There is scant population-based information on functional outcome, survival, and recurrence for ischemic stroke subtypes. METHODS: We identified all residents of Rochester, Minnesota, with a first ischemic stroke from 1985 through 1989 using the resources of the Rochester Epidemiology Project medical records linkage system. After reviewing medical records and imaging studies, we assigned patients to 4 major ischemic stroke categories based on National Institute of Neurological Diseases and Stroke Data Bank criteria: large vessel cervical or intracranial atherosclerosis with stenosis (ATH, n=74), cardioembolic (CE, n=132), lacunar (LAC, n=72), and infarct of uncertain cause (IUC, n=164). We used the Rankin disability score to assess functional outcome and the Kaplan-Meier product-limit method and Cox proportional hazards regression analysis with bootstrap validation to estimate rates and identify predictors of survival and recurrent stroke among these patients. RESULTS: Rankin disabilities were different across stroke subtypes at the time of stroke and 3 months and 1 year later (P=0.001). LAC was associated with milder deficits compared with other subtypes. Mean follow-up among the 442 patients in the cohort was 3.2 years. Estimated rates of recurrent stroke at 30 days were significantly different (P<0.001): ATH, 18.5% (95% CI 9.4% to 27.5%); CE, 5.3% (95% CI 1.2% to 9.6%); LAC, 1.4% (95% CI 0.0% to 4.1%); and IUC, 3. 3% (95% CI 0.4% to 6.2%). After adjusting for age, sex, and stroke severity, infarct subtype was an independent determinant of recurrent stroke within 30 days (P=0.0006; eg, risk ratio for ATH compared with CE=3.3, 95% CI 1.2 to 9.3) but not long term (P=0.07). Four of 25 recurrent strokes within 30 days were procedure-related, each in patients with ATH. Five-year death rates were significantly different (P<0.001): ATH, 32.2% (95% CI 21.1% to 43.2%); CE, 80.4% (95% CI 73.1% to 87.6%); LAC, 35.1% (95% CI 23.6% to 46.0%); and IUC, 48.6% (95% CI 40.5% to 56.7%). With adjustment for age, sex, cardiac comorbidity, and stroke severity, the subtype of ischemic stroke was an independent determinant of long-term (P=0.018; eg, risk ratio for ATH compared with cardioembolic=0.47, 95% CI 0.29 to 0.77) but not 30-day survival (P=0.2). CONCLUSIONS: Early recurrence rates for ischemic stroke caused by ATH are higher than those for other subtypes and higher than previous non-population-based studies have reported. Some of the increased risk of early recurrence among patients with ATH may be iatrogenic. Patients with LAC have better poststroke functional status than those with other subtypes. Survival is poorest among those with ischemic stroke with a cardiac source of embolism. PMID- 10797167 TI - Association between high homocyst(e)ine and ischemic stroke due to large- and small-artery disease but not other etiologic subtypes of ischemic stroke. AB - BACKGROUND AND PURPOSE: Elevated plasma homocyst(e)ine may be a causal and modifiable risk factor for ischemic stroke, but the results of previous studies have been conflicting. One possible explanation is that homocyst(e)ine may only be associated with certain pathophysiological subtypes of ischemic stroke. METHODS: We conducted a case-control study of 219 hospital cases with a first ever ischemic stroke and 205 randomly selected community control subjects stratified by age, sex, and postal code. With the use of established criteria, cases of stroke were classified by etiologic subtype in a blinded fashion. The prevalence of conventional vascular risk factors, fasting plasma homocyst(e)ine levels, vitamin levels, and nucleotide 677 methylene tetrahydrofolate reductase (MTHFR) genotypes were determined in cases and controls. RESULTS: Increasing homocyst(e)ine was a strong and independent risk factor for ischemic stroke (adjusted OR 2.7, 95% CI 1.4 to 5.1 for a 5-micromol/L increase in fasting plasma homocyst(e)ine from 10 to 15 micromol/L). Compared with the lowest quartile, the highest quartile of homocyst(e)ine was associated with an adjusted OR of ischemic stroke of 2.2 (95% CI 1.1 to 4.2). Mean plasma homocyst(e)ine was significantly higher in cases of ischemic stroke due to large-artery disease (14.1 micromol/L, 95% CI 12.5 to 15.9, P<0.001) and small-artery disease (12.7 micromol/L, 95% CI 11. 4 to 14.1, P=0.004) compared with control subjects (10.5 micromol/L; 95% CI 10.0 to 11.0) but not in cardioembolic or other etiologic subtypes of ischemic stroke. Compared with the lowest quartile of homocyst(e)ine, the upper 3 quartiles were associated with an adjusted OR of ischemic stroke due to large artery disease of 3.0 (95% CI 0.8 to 10.8) for the second quartile, 5.6 (95% CI 1.6 to 20) for the third quartile, and 8.7 (95% CI 2.4 to 32) for the fourth quartile (P for trend=0.0005). However, despite a clear association between the TT MTHFR genotype and elevated fasting plasma homocyst(e)ine, there was no association between MTHFR genotype and ischemic stroke or subtype of ischemic stroke. CONCLUSIONS: There is a strong, graded association between increasing plasma homocyst(e)ine and ischemic stroke caused by large-artery atherosclerosis and, to a much lesser extent, small-artery disease, but not cardioembolic or other etiologic subtypes of ischemic stroke. Our results are consistent with the hypothesis that the deleterious effect of high homocyst(e)ine is mediated primarily via a proatherogenic effect. PMID- 10797168 TI - Verifying the stroke-free phenotype by structured telephone interview. AB - BACKGROUND AND PURPOSE: Case-control, cohort, and twin studies support a genetic contribution to ischemic stroke risk. Sibling pair linkage methods require identification of concordant or discordant siblings or both. We designed and tested a structured telephone interview to verify the stroke-free phenotype. METHODS: A coordinator unaware of medical record data used an 8-item questionnaire to conduct a structured telephone interview of 70 outpatients aged >60 years. The questionnaire inquired about the sudden onset of deficits in strength, sensation, vision, and language. A subject was defined as stroke free by interview if responses to all items on the questionnaire were negative. Results of the telephone interview were compared with data obtained from a systematic medical record review (benchmark). RESULTS: Interview time was 5 minutes or less for all subjects. All subjects who began the interview completed it. Records were reviewed in all subjects. Medical record review detected ischemic stroke or transient ischemic attack (TIA), or both, in 5 patients (7%). There were no significant differences in sex distribution or risk factor rates in patients who were designated stroke free or not stroke free by interview. Having 1 or more positive items on the questionnaire was significantly associated with finding stroke (P<0.001), TIA (P<0.001), or either stroke or TIA (P<0.001), on medical record review. The telephone interview had a sensitivity of 1.0 (95% CI 0.48 to 1.0), specificity of 0.86 (95% CI 0.75 to 0.93), positive predictive value of 0.36 (95% CI 0.13 to 0.65), and negative predictive value of 1.0 (95% CI 0.94 to 1.0). CONCLUSIONS: Our instrument can identify the stroke-free individual with a high degree of confidence in a very efficient manner. It may be particularly suited for centralized verification of stroke discordancy in multicentered sib-pair genetic studies. PMID- 10797169 TI - Impact on stroke subtype diagnosis of early diffusion-weighted magnetic resonance imaging and magnetic resonance angiography. AB - BACKGROUND AND PURPOSE: The purpose of the present study was to assess the diagnostic usefulness of early diffusion-weighted MRI (DWI) and MR angiography (MRA) in patients with ischemic stroke. Past approaches to stroke diagnosis required a series of diagnostic tests over several days of hospitalization. New magnetic resonance methodologies that include DWI and MRA may allow more rapid characterization of stroke pathophysiology. However, no previous study has assessed the impact on formal stroke subtype diagnosis of early imaging with DWI/MRA. METHODS: We analyzed 46 consecutive patients with acute ischemic stroke who underwent DWI/MRA within 24 hours of admission. Initial diagnoses were rendered with use of the 2 most widely used formal stroke subtype classification schemes, the TOAST and the Oxfordshire methods, which were applied to patients after CT/conventional MRI but before DWI/MRA. Modified TOAST and Oxfordshire diagnoses were then rendered based on the results of day 1 DWI, MRA, and DWI plus MRA. Final TOAST/Oxfordshire diagnoses at discharge were taken as the gold standard. RESULTS: Compared with final diagnoses, pre-MRI TOAST diagnoses matched final diagnoses in 48%, improving to 83% after DWI alone, 56% after MRA alone, and 94% after DWI plus MRA. For the TOAST diagnostic subtypes of large-vessel atherothromboembolism and small-vessel disease, pre-MRI diagnoses matched final diagnoses in 56% and 35% of patients, respectively, improving to 89% and 100% after DWI/MRA. Pre-MRI Oxfordshire diagnoses matched final diagnoses in 67% of patients, improving to 100% after DWI. CONCLUSIONS: The use of DWI/MRA within 24 hours of hospitalization substantially improves the accuracy of the diagnosis of early ischemic stroke subtype. When initial management and clinical trial eligibility decisions are influenced by stroke subtype, day 1 multimodal MRI is advantageous as a guide to therapy. PMID- 10797171 TI - Combined diffusion-weighted and perfusion-weighted flow heterogeneity magnetic resonance imaging in acute stroke. AB - BACKGROUND AND PURPOSE: The heterogeneity of microvascular flows is known to be an important determinant of the efficacy of oxygen delivery to tissue. Studies in animals have demonstrated decreased flow heterogeneity (FH) in states of decreased perfusion pressure. The purpose of the present study was to assess microvascular FH changes in acute stroke with use of a novel perfusion-weighted MRI technique and to evaluate the ability of combined diffusion-weighted MRI and FH measurements to predict final infarct size. METHODS: Cerebral blood flow, FH, and plasma mean transit time (MTT) were measured in 11 patients who presented with acute (<12 hours after symptom onset) stroke. Final infarct size was determined with follow-up MRI or CT scanning. RESULTS: In normal brain tissue, the distribution of relative flows was markedly skewed toward high capillary flow velocities. Within regions of decreased cerebral blood flow, plasma MTT was prolonged. Furthermore, subregions were identified with significant loss of the high-flow component of the flow distribution, thereby causing increased homogeneity of flow velocities. In parametric maps that quantify the acute deviation of FH from that of normal tissue, areas of extreme homogenization of capillary flows predicted final infarct size on follow-up scans of 10 of 11 patients. CONCLUSIONS: Flow heterogeneity and MTT can be rapidly assessed as part of a routine clinical MR examination and may provide a tool for planning of individual stroke treatment, as well as in targeting and evaluation of emerging therapeutic strategies. PMID- 10797170 TI - Diffusion- and perfusion-weighted MRI patterns in borderzone infarcts. AB - BACKGROUND AND PURPOSE: The pathophysiology of borderzone infarcts is not well understood. We investigated whether combined diffusion-weighted imaging (DWI) and perfusion-weighted imaging (PWI) could identify pathophysiologically meaningful categories of borderzone infarcts. METHODS: Seventeen patients with borderzone infarcts were identified from the Beth Israel Deaconess Medical Center Stroke Database. All patients had DWI and PWI, the majority of them within the first 24 hours of symptom onset. RESULTS: Three patterns of perfusion abnormalities were associated with the diffusion lesions: 1, normal perfusion (5 patients); 2, localized perfusion deficits matching the area of restricted diffusion (5 patients); and 3, extensive perfusion deficits involving 1 or more vascular territories (7 patients). All but 1 patient with pattern 1 had transient peri infarct hypotension as the presumed stroke mechanism. Two patients with pattern 2 had cardiac or aortic embolic sources; none had large-artery disease or arterial hypotension. Reperfusion was detected in all patients with this pattern who submitted to a follow-up study. All patients with pattern 3 had severe stenosis or occlusion of a large artery: the internal carotid, anterior cerebral, or middle cerebral. CONCLUSIONS: We postulate that the perfusion abnormality varies according to the mechanism of the borderzone infarction. Transient perfusion deficits occurring with hypotension in the absence of significant large-artery disease may not be revealed by PWI. Embolism may cause some cases of small borderzone perfusion deficits. Critical large-artery disease may cause large territorial perfusion deficits and predispose to borderzone infarction. PMID- 10797172 TI - Reproducibility validation study comparing analog and digital imaging technologies for the measurement of intima-media thickness. AB - BACKGROUND AND PURPOSE: New advances in B-mode imaging technologies have led to improved quality in the detection of minute changes in the surface of intima media thickness (IMT) and plaques. The new digital systems, with increased numbers of imaging channels, multiple frequency probes, and increased microprocessing speeds, now generate images comparable to those of the analog predecessors. Can these digital systems have reproducibility comparable to that of a pure analog system? We compared the Biosound 2000II (analog) system with the Esaote AU4 (digital) system. METHODS: Twenty-two subjects were chosen who had varying degrees of IMT on the far wall of the common carotid artery. Common carotid IMT was determined twice: the first time with the analog system and the second time with the digital system. With each system, replicate scans were made within 2 weeks. RESULTS: The intramethod agreement was high with the analog system, with a bias between readings of -0.010+/-0.033 mm, mean absolute difference of 0.027+/-0.020 mm, repeatability coefficient of 0.067, and correlation coefficient of 0.97. The digital system provided the highest reproducibility with a bias between readings of 0.002+/-0.016 mm, mean absolute difference of 0.012+/-0.011 mm, repeatability coefficient of 0.033, and correlation coefficient of 0. 99. When the analog and digital systems were compared, the bias between readings was -0.011+/-0.024 mm with good agreement between the 2 systems; the repeatability coefficient was 0.047, with all points within +/-2 SDs of the mean difference. The mean absolute difference between the 2 measurements was 0.018+/-0.015 mm with a correlation coefficient of 0.98. CONCLUSIONS: The digital system for IMT evaluation compares well with the more widely used analog system and provides a reliable technology for common carotid IMT measurement that can be applied to clinical trials. PMID- 10797174 TI - Effect of short-term hyperventilation on cerebral blood flow autoregulation in patients with acute bacterial meningitis. AB - BACKGROUND AND PURPOSE: Cerebral blood flow (CBF) autoregulation is impaired in patients with acute bacterial meningitis: this may be caused by cerebral arteriolar dilatation. We tested the hypothesis that CBF autoregulation is recovered by acute mechanical hyperventilation in 9 adult patients with acute bacterial meningitis. METHODS: Norepinephrine was infused to increase mean arterial pressure (MAP) 30 mm Hg from baseline. Relative changes in CBF were concomitantly recorded by transcranial Doppler ultrasonography of the middle cerebral artery, measuring mean flow velocity (V(mean)), and by measurement of arterial to jugular oxygen content difference (a-v DO(2)). The slope of the regression line between MAP and V(mean) was calculated. Measurements were performed during normoventilation and repeated after 30 minutes of mechanical hyperventilation. RESULTS: At normoventilation (median PaCO(2) 4.4 kPa, range 3.5 to 4.9), MAP was increased from 68 mm Hg (60 to 101) to 109 mm Hg (95 to 126). V(mean) increased with MAP from 48 cm/s (30 to 61) to 65 cm/s(33 to 86) (P<0.01), and a-v DO(2) decreased from 2.2 mmol/L (1.0 to 2.7) to 1.4 mmol/L (0.8 to 1.8) (P<0.05). During hyperventilation (PaCO(2) 3.5 kPa, range 3.3 to 4.1), MAP was increased from 76 mm Hg (58 to 92) to 109 mm Hg (95 to 121). V(mean) increased from 45 cm/s (29 to 55) to 53 cm/s (33 to 78) (P<0.01), and a-v DO(2) decreased from 2.5 mmol/L (1.8 to 3.0) to 1.8 mmol/L (1.2 to 2.4) (P<0.05). Four patients recovered autoregulation completely during hyperventilation. The slope of the autoregulation curve decreased during hyperventilation compared with normoventilation (P<0.05). CONCLUSIONS: CBF autoregulation is partially recovered during short-term mechanical hyperventilation in patients with acute bacterial meningitis, indicating that cerebral arteriolar dilation in part accounts for the regulatory impairment of CBF in these patients. PMID- 10797173 TI - Arterial pulsatility as an index of cerebral microangiopathy in diabetes. AB - BACKGROUND AND PURPOSE: This study was designed to evaluate cerebral hemodynamic changes related to diabetes mellitus (DM) with transcranial Doppler ultrasonography (TCD). METHODS: We measured the flow velocities and the Gosling pulsatility index (PI) of the middle cerebral artery (MCA), extracranial internal carotid artery (ICA), and basilar artery (BA) in 56 stroke-free, normotensive patients with type 2 DM and 70 age- and gender-matched healthy volunteers. Patients were divided into 2 groups according to the presence of microvascular complications such as retinopathy, nephropathy, and neuropathy. RESULTS: Patients showed slightly lower hematocrit and higher serum fibrinogen levels than control subjects, but other clinical profiles, including stroke risk factors except for diabetes, were comparable between patients and controls. The flow velocity of the ICA but not the MCA and BA in patients regardless of the complication was significantly higher than that in controls. The PIs of the MCA and ICA were significantly higher in patients with complication than those without complication, as well as in controls. The PI of the BA was also significantly higher, even in patients without complication, than in controls. The PIs of the MCA and ICA but not the BA were closely correlated with the duration of DM (r(2)=0.46 and 0.34, respectively). CONCLUSIONS: This study defines TCD findings of diabetes-related cerebral hemodynamic changes and suggests that the PI reflects microangiopathic changes of cerebral vessels. PMID- 10797175 TI - Visualization of the basilar artery by transcranial color-coded duplex sonography : comparison with postmortem results. AB - BACKGROUND AND PURPOSE: Transcranial color-coded sonography (TCCS) via the suboccipital approach allows direct and continuous visualization of the basilar artery (BA). In this study, we intended to evaluate the ability of native TCCS in visualizing the length of the BA by means of a comparison with postmortem measurements. METHODS: The BA was prospectively studied by TCCS shortly before death (median 3 days) in 46 moribund neurological patients (mean+/-SD age 71.1+/ 13.1 years). The length of the BA was determined by measuring the distance between the vertebrobasilar junction and the deepest available flow signal in the top of the BA. During autopsy, photos of the vertebrobasilar system were taken to evaluate the true anatomic length and variations of the course of BA in situ, eg, straight, curved, or S-shaped. RESULTS: Comparison of the in vivo ultrasound measurements of BA length and postmortem data was possible in 44 of 46 cases. In the 2 remaining patients, the BA was occluded. The mean insonation depth of the vertebrobasilar junction was found at 66.9+/-7.1 mm. The mean BA length was 21.5+/-6. 8 mm by color-coded duplex and 32.9+/-6 mm anatomically (P<0.0001). The mean difference between color mode and anatomic findings was 11. 3+/-6.4 mm in the case of a straight BA (35 cases) and 16.3+/-4.8 mm in an anatomically tortuous course of the BA (9 cases). CONCLUSIONS: Color duplex imaging enables correct visualization of the proximal two thirds of the BA, but only exceptionally of its distal one third. A tortuous course of the BA leads to an underestimation of its anatomic length. PMID- 10797176 TI - Transcranial doppler ultrasound criteria for recanalization after thrombolysis for middle cerebral artery stroke. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) can demonstrate arterial occlusion and subsequent recanalization in acute ischemic stroke patients treated with intravenous tissue plasminogen activator (tPA). Limited data exist to assess the accuracy of recanalization by TCD criteria. METHODS: In patients with acute middle cerebral artery (MCA) occlusion treated with intravenous tPA, we compared posttreatment TCD with angiography (digital subtraction or magnetic resonance). On TCD, complete occlusion was defined by absent or minimal signals, partial occlusion by blunted or dampened signals, and recanalization by normal or stenotic signals. Angiography was evaluated with the Thrombolysis In Myocardial Ischemia (TIMI) grading scale. RESULTS: Twenty-five patients were studied (age 61+/-18 years, 16 men and 9 women). TCD was performed at 12+/-16 hours and angiography at 41+/-57 hours after stroke onset, with 52% of studies performed within 3 hours of each other. Recanalization on TCD had the following accuracy parameters compared with angiography: sensitivity 91%, specificity 93%, positive predictive value (PPV) 91%, and negative predictive value (NPV) 93%. To predict partial occlusion (TIMI grade II), TCD had sensitivity of 100%, specificity of 76%, PPV of 44%, and NPV of 100%. TCD predicted the presence of complete occlusion on angiography (TIMI grade 0 or I) with sensitivity of 50%, specificity of 100%, PPV of 100%, and NPV of 75%. TCD flow signals correlated with angiographic patency (chi(2)=24.2, P<0.001). CONCLUSIONS: Complete MCA recanalization on TCD accurately predicts angiographic findings. Although a return to normal flow dynamics on TCD was associated with complete angiographic resumption of flow, partial signal improvement on TCD corresponded with persistent occlusion on angiography. PMID- 10797177 TI - Trials of thrombolysis in acute ischemic stroke: does the choice of primary outcome measure really matter? AB - BACKGROUND AND PURPOSE: Controversy regarding the risks and benefits of thrombolysis has not been helped by the perception that some trials were "positive" and others "negative" on their primary outcome measure of either "good" or "poor" functional outcome. We wondered whether the definition of good or poor functional outcome might have contributed to this perception, and what effect altering the definition might have on the individual trials and on the systematic review of all the trials combined. METHODS: We analyzed data on functional outcome, extracted from the randomized trials of thrombolysis in acute ischemic stroke, according to good (modified Rankin scale scores of 0 to 1 versus 2 to 6) and poor (modified Rankin 0 to 2 versus 3 to 6) functional outcome, to determine the effects of thrombolysis. RESULTS: Twelve trials (4342 patients, treated up to 6 hours after stroke) contributed to this analysis. Overall, there was no difference in the estimate of treatment effect between the 2 definitions (modified Rankin 0 to 2 versus 3 to 6, and 0 to 1 versus 2 to 6 [ORs 0.83 and 0.79, respectively]). However, the apparent "success" of several individual trials did alter. CONCLUSIONS: We should not place undue emphasis on the results of individual trials, when a change of a single point on the Rankin scale can make the difference between "success" and "failure." Overall, by either analysis, there was a significant benefit in patients treated with thrombolysis up to 6 hours after stroke. PMID- 10797178 TI - Regional myocardial perfusion after experimental subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The pathophysiology of cardiac injury after subarachnoid hemorrhage (SAH) remains controversial. Data from animal models suggest that catecholamine-mediated injury is the most likely cause of cardiac injury after SAH. However, researchers also have proposed myocardial ischemia to be the underlying cause, as a result of coronary artery disease, coronary artery spasm, or hypertension and tachycardia. To test the hypothesis that SAH-induced cardiac injury occurs in the absence of myocardial hypoperfusion, we developed an experimental canine model that reproduces the clinical and pathological cardiac lesions of SAH and defines the epicardial and microvascular coronary circulation. METHODS: Serial ECG, hemodynamic measurements, coronary angiography, regional myocardial blood flow measurements by radiolabeled microspheres, 2D echocardiography, and myocardial contrast echocardiography were performed in 9 dogs with experimental SAH and 5 controls. RESULTS: Regional wall motion abnormalities were identified in 8 of 9 SAH dogs and 1 of 5 controls (Fisher's Exact Test, P=0.02) but no evidence was seen of coronary artery disease or spasm by coronary angiography and of significant myocardial hypoperfusion by either regional myocardial blood flow or myocardial contrast echocardiography. CONCLUSIONS: In this experimental model of SAH, a unique form of regional left ventricular dysfunction occurs in the absence of myocardial hypoperfusion. Future studies are justified to determine the cause of cardiac injury after SAH. PMID- 10797179 TI - Early exclusive use of the affected forelimb after moderate transient focal ischemia in rats : functional and anatomic outcome. AB - BACKGROUND AND PURPOSE: Previous work by researchers in our laboratory has shown that in the rat, the exclusive use of the affected forelimb during an early critical period exaggerates lesion volume and retards functional recovery after electrolytic lesions of the forelimb sensorimotor cortex. In the present study, we examined the effects of exclusive use of the affected forelimb after middle cerebral artery occlusion (MCAO). METHODS: Ischemia of moderate severity was produced in male Long-Evans rats through 45 minutes of occlusion of the left middle cerebral and both common carotid arteries. Exclusive use of either the affected or unaffected forelimb was forced through immobilization of either the ipsilateral (MCAO+ipsi) or contralateral (MCAO+contra) forelimb, respectively, for 10 days in a plaster cast, or the animal was left uncasted (MCAO+nocast). Sham surgeries were performed, and animals were also casted for 10 days or left uncasted. Sensorimotor testing was performed during days 17 to 38. At the end of sensorimotor testing, cognitive performance was tested with use of the Morris water maze. In a separate experiment, temperatures and corticosterone levels were measured during the 10-day period after 45-minute ischemia and casting. RESULTS: The MCAO+ipsi group performed worse on sensorimotor tasks than the MCAO+contra, MCAO+nocast, and sham groups. Infarct volume was significantly larger in the MCAO+ipsi group than in the sham and MCAO+contra groups but not in the MCAO+nocast group. No group differences were found with the Morris water maze, and no group differences were found in either temperature or plasma corticosterone level. CONCLUSIONS: The exclusive use of the affected forelimb immediately after focal ischemia has detrimental effects on sensorimotor function that cannot be attributed to hyperthermia or stress. PMID- 10797180 TI - Leukocyte accumulation and hemodynamic changes in the cerebral microcirculation during early reperfusion after stroke. AB - BACKGROUND AND PURPOSE: Leukocytes contribute to cerebral ischemia-reperfusion injury. However, few experimental models examine both in vivo behavior of leukocytes and microvascular rheology after stroke. The purpose of the present study was to characterize patterns of leukocyte accumulation in the cerebral microcirculation and to examine the relationship between leukocyte accumulation and microcirculatory hemodynamics after middle cerebral artery occlusion and reperfusion (MCAO-R). METHODS: Male rats (250 to 350 g) were anesthetized and ventilated. Tail catheters were inserted for measurement of arterial blood gases and administration of drugs. Body temperature was maintained at 37 degrees C. Animals were subjected to 2 hours of MCAO by the filament method. A cranial window preparation was performed, and the brain was superfused with warm, aerated artificial cerebrospinal fluid. Reperfusion was initiated by withdrawing the filament, and the pial microcirculation was observed by use of intravital fluorescence microscopy. Leukocyte accumulation in venules, arterioles, and capillaries; leukocyte rolling in venules; and leukocyte venular shear rate were assessed during 1 hour of reperfusion. RESULTS: We found significant leukocyte adhesion in cerebral venules during 1 hour of reperfusion after 2 hours of MCAO. Leukocyte trapping in capillaries and adhesion to arterioles after MCAO-R tended to increase compared with controls, but the increase was not significant. We also found that shear rate was significantly reduced in venules during early reperfusion after MCAO. CONCLUSIONS: A model using the filament method of stroke and fluorescence microscopy was used to examine white-cell behavior and hemodynamics in the cerebral microcirculation after MCAO-R. We observed a significant increase in leukocyte rolling and adhesion in venules and a significant decrease in blood shear rate in the microcirculation of the brain during early reperfusion. Leukocytes may activate and damage the blood vessels and surrounding brain cells, which contributes to an exaggerated inflammatory component to reperfusion. The model described can be used to examine precisely blood cell-endothelium interactions and hemodynamic changes in the microcirculation during postischemic reperfusion. Information from these and similar experiments may contribute to our understanding of the early inflammatory response in the brain during reperfusion after stroke. PMID- 10797181 TI - The corticosterone synthesis inhibitor metyrapone prevents hypoxia/ischemia induced loss of synaptic function in the rat hippocampus. AB - BACKGROUND AND PURPOSE: Ischemia is accompanied by abundant corticosterone secretion, which could potentially exacerbate brain damage via activation of glucocorticoid receptors. We addressed whether manipulating steroid levels during ischemia affects hippocampal synaptic function along with neuronal structure. Moreover, we established whether pretreatment with the glucocorticoid receptor antagonist RU38486 is as effective in preventing deleterious effects after ischemia as is the steroid synthesis inhibitor metyrapone. METHODS: Rats underwent 20 minutes of unilateral hypoxia/ischemia (HI). Convulsions were monitored after HI, and 24 hours later, field potentials were recorded in vitro in the hippocampal CA1 area in response to stimulation of the Schaffer collateral/commissural fibers. Morphological alterations were determined in brain slices from the same animals. Data were correlated with steroid treatment before HI. RESULTS: Metyrapone suppressed plasma corticosteroid levels during HI, whereas corticosterone treatment significantly elevated plasma steroid levels. These treatments affected the incidence of visible seizures after HI: corticosterone treatment resulted in the highest incidence, whereas metyrapone attenuated the occurrence of seizures. Moreover, the HI-induced impairment in synaptic transmission in the CA1 area in vitro was exacerbated by concomitant corticosteroid treatment and alleviated by pretreatment with metyrapone. In parallel, degenerative changes in the hippocampus after HI were most pronounced after corticosterone treatment, whereas metyrapone reduced these alterations. RU38486 was effective only in reducing the incidence of seizures shortly after ischemia. CONCLUSIONS: We tentatively conclude that synaptic function along with cellular integrity is preserved after HI by preventing the ischemia-evoked rise in corticosteroid levels rather than blocking the glucocorticoid receptor. PMID- 10797182 TI - Progesterone exacerbates striatal stroke injury in progesterone-deficient female animals. AB - BACKGROUND AND PURPOSE: We have previously shown that female animals experience substantial protection from brain injury after reversible middle cerebral artery occlusion (MCAO) compared with their male or ovariectomized female counterparts. The reproductive steroid estrogen has been shown to provide neuroprotection from a variety of experimental insults, but the importance of progesterone as an anti ischemic treatment has not been well explored. We evaluated histological outcomes after MCAO in ovariectomized female rats with or without acute or chronic progesterone replacement therapy. METHODS: Age-matched, adult female Wistar rats were ovariectomized and treated with 0, 30, or 60 mg/kg progesterone IP 30 minutes before ischemia (n=12 to 14 per group) or with 30 mg/kg progesterone IP daily for 7 to 10 days before ischemia (n=16). Each animal subsequently underwent 2 hours of MCAO with the intraluminal filament technique, followed by 22 hours of reperfusion. Ipsilateral parietal cortex perfusion was monitored with laser Doppler flowmetry throughout ischemia. Cortical, caudate-putamen, and hemispheric infarction volumes were determined with 2,3,5-triphenyltetrazolium chloride staining and digital image analysis. RESULTS: Intraischemic plasma progesterone levels were 5+/-3, 102+/-20,* 181+/-28,* and 133+/-25* ng/mL in the 0, 30, and 60 mg/kg acute progesterone group and the 30 mg/kg chronic progesterone group, respectively (*P<0.05 compared with 0 mg/kg). Caudate-putamen infarction volume (percent contralateral structure) was significantly increased by chronic progesterone treatment: 45.6+/-5.1%* in the 30 mg/kg chronic progesterone group and 29.2+/-5.3%, 35.8+/-5.1%, and 42.0+/-5.0% in the 0, 30, and 60 mg/kg acute progesterone groups, respectively (*P<0.05 compared with 0 mg/kg). Cortical and total hemispheric infarction volumes (percent contralateral structure) were unchanged by progesterone treatment. CONCLUSIONS: Exogenous progesterone therapy does not ameliorate histological injury after MCAO in previously ovariectomized, adult female rats. Furthermore, chronic progesterone administration can exacerbate infarction in subcortical regions. PMID- 10797183 TI - Hypertonic mannitol loading of NF-kappaB transcription factor decoys in human brain microvascular endothelial cells blocks upregulation of ICAM-1. AB - BACKGROUND AND PURPOSE: An acute inflammatory response exacerbates tissue injury during acute ischemic stroke. The transcription factor nuclear factor (NF)-kappaB plays a key role in endothelial cell activation and the inflammatory response. Targeted genetic disruption of NF-kappaB activation in cerebral endothelial cells may be protective in stroke. We determined whether a NF-kappaB transcription factor decoy (TFD) could block intercellular adhesion molecule (ICAM)-1 upregulation, an indicator of endothelial cell activation. METHODS: We modeled ischemia-reperfusion in vitro by exposing cultured human brain microvascular endothelial cells (HBMEC) to tumor necrosis factor (TNF)-alpha and conditions of hypoxia-reoxygenation (H/R). Mannitol was used to load phosphothiorated oligonucleotides containing 3 copies of the kappaB binding sequences (TFDs) into cultured HBMEC. An NF-kappaB TFD, a mutated NF-kappaB TFD, and a scrambled TFD were studied for their effect on ICAM-1 mRNA levels and surface ICAM-1 by ELISA. RESULTS: Hyperosmolar loading with mannitol permitted rapid transfection of TFD into endothelial cell nuclei. The NF-kappaB TFD but not the mutated or scrambled TFD competed with a kappaB sequence for binding to nuclear extracts from HBMEC exposed to TNF-alpha. The NF-kappaB TFD blocked the TNF-alpha-induced and H/R induced increase in ICAM-1 mRNA levels and the upregulation of surface ICAM-1. CONCLUSIONS: Mannitol delivers phosphothiorated oligonucleotides into cultured HBMEC. An NF-kappaB decoy blocks both TNF-alpha-induced and H/R-induced ICAM-1 upregulation in HBMEC. Targeted genetic disruption of endothelial NF-kappaB activation may be of benefit in acute ischemic stroke. PMID- 10797184 TI - Solitary tract nuclei in acute heart failure. AB - BACKGROUND AND PURPOSE: Symmetrical necrosis of the brain stem nuclei has been described as a consequence of severe transitory cerebral hypoxia mainly in neonates or young adults who experienced an episode of acute ischemia due to transitory acute heart failure. We report selective bilateral lesions of the solitary tract nuclei in 5 adults with short survival intervals after acute heart failure. METHODS: In 5 patients who died due to cardiovascular pathology, histological examination was performed on multiple samples of cerebral hemispheres, on transverse sections of the midbrain and pons, and on transverse serial sections of the medulla stained with hematoxylin-eosin, Kluver-Barrera, and Luxol fast blue. The 3-dimensional reconstruction of the extension and topography of the medullary lesions was obtained with computed image analysis. RESULTS: In 4 subjects who died soon after an episode of acute heart failure (range of survival 10 hours to 2 days), the dorsal portion of the solitary tract nuclei showed an eosinophilic roundish aspect bilaterally. In their context, the neurons showed changes characteristic of ischemic coagulation necrosis. In a fifth patient, a 32-year-old man who died 15 days after an episode of cardiac arrest, 2 circumscribed symmetrical infarcts with macrophagic and astrocytic reactions were found at the same level. The topography of the lesions and the inflammatory reaction and gliosis of patient 5 suggest that the findings in the other 4 patients correspond to initial features of selective lesions of the solitary tract nuclei after acute heart failure: the short interval of survival prevented the evolution of the reactive process. The nucleus is localized at the watershed zone between the terminal branches of the medullary collateral vessels of the vertebral arteries, thus representing the last meadow in the case of sudden fall of the systemic blood flow due to acute heart failure. The absence of lesions of other medullary and pontine nuclei accounts for a selective vulnerability of the neurons of the solitary tract nuclei, and the selective dendritic lesions suggest an excitotoxic component to ischemic cell death. CONCLUSIONS: The commonly accepted resistance of the medullary centers to ischemic hypoxia in adults apparently could be due to the rapidity of death, which prevents the evolution of lesions that can be diagnosed. In addition, minor lesions in the medullary tegmentum after acute heart failure could play a role in the prevention of the resumption of autonomous cardiac and respiratory functions despite life-saving procedures. PMID- 10797185 TI - Lipoprotein(a) and the risk of stroke. PMID- 10797186 TI - Intrathecal sodium nitroprusside improves cerebral blood flow and oxygenation in refractory cerebral vasospasm and ischemia in humans. PMID- 10797187 TI - The relative contributions of the anterior and posterior circulations to global cerebral blood flow. PMID- 10797188 TI - Failure of standards in reporting the composition of artificial cerebral spinal fluid in studies of the pial blood vessels. PMID- 10797190 TI - 25th international stroke conference : (Corrections in bold) PMID- 10797189 TI - Abstracts of literature PMID- 10797191 TI - Lumbar Spine Imaging: Role in Clinical Decision Making. AB - Imaging studies are an integral component of the evaluation of the lumbar spine. For each study there is a specific role, an appropriate indication, and a correct time for utilization during the course of a patient's illness. The physician must know the specificity and sensitivity of each test, as well as the prevalence of abnormal findings in asymptomatic persons, to properly order and interpret the results of the studies. Many errors in decision making arise, not from misinterpretation of what is seen on imaging studies, but rather from misuse of imaging information in the clinical decision making process. Because all neurodiagnostic imaging modalities reveal abnormalities in at least a third of asymptomatic persons, the use of these tests for general screening is dangerous. The challenge for the future is to be able to better correlate what is seen on imaging studies with the patient's symptoms. Obtaining these expensive studies too early in the treatment of self-limited disorders is costly and often misleading for both the physician and the patient. The authors outline an approach to the judicious timing of imaging studies and discuss pitfalls in their interpretation in the evaluation of degenerative diseases of the lumbar spine. PMID- 10797192 TI - Tibial Nonunion: Treatment Alternatives. AB - Because the spectrum of injuries to the tibia is so great, no single method of treatment is applicable to all nonunions. Therefore, it is important for surgeons who treat tibial nonunions to be skilled in several different methods of treatment. In patients with significant deformities, electrical stimulation, isolated fibular osteotomy, and bone grafts alone are unsatisfactory treatment options. In aseptic nonunions, the use of intramedullary nailing or compression plating appears to have many advantages. In previously closed and selected grade I and grade II open fractures, reamed intramedullary nailing is a safe and effective method of treatment. Because of the risk of infection, reamed nailing is not recommended after external fixation of open fractures. In these cases as well as others, the authors prefer plate osteosynthesis. With few exceptions, the plate should be placed, under tension, on the convex side of the tibia. Used in this fashion, the plate can assist in correction of any deformity and can also provide stable internal fixation. Half-pin external fixation is used primarily in the management of infected fractures. Ilizarov and other small-wire circular fixators have proved effective in treating complex-composite deformities associated with sepsis, bone loss, shortening, angulation, or malrotation. Amputation may be warranted if a functional limb cannot be achieved. PMID- 10797193 TI - Trochanteric Osteotomy for Total Hip Arthroplasty: Six Variations and Indications for Their Use. AB - Trochanteric osteotomy of the femur in total hip replacement used to be a simple, stereotyped technique and was declining in frequency of use. Many variations in the technique have evolved recently, including the trochanteric slide and the extended trochanteric osteotomy, which have increased the flexibility and utility of trochanteric osteotomy and the frequency of its application. The authors describe six trochanteric osteotomies and discuss the indications for their use. PMID- 10797194 TI - Acute and Chronic Traumatic Injuries of the Sternoclavicular Joint. AB - Acute and chronic traumatic injuries of the sternoclavicular joint require accurate diagnosis and management if complications are to be avoided. Sternoclavicular subluxation or dislocation, medial clavicle physeal injuries, and degenerative arthritis are the most frequently diagnosed of these relatively uncommon injuries. The medial clavicular epiphysis does not ossify until the 18th to 20th year. Knowledge of its developmental anatomy is essential because most physeal injuries will heal with time without surgical intervention. In contrast, posterior dislocation of the sternoclavicular joint requires prompt closed or open reduction, as posterior displacement of the medial clavicle has been associated with numerous complications, including respiratory distress, venous congestion or arterial insufficiency, brachial plexus compression, and myocardial conduction abnormalities. A myriad of procedures have been recommended for repair or reconstruction of the sternoclavicular joint. On the basis of the authors' experience and review of the literature, they advocate surgical resection of the medial clavicle, with maintenance, repair, or reconstruction of the costoclavicular ligaments, when surgery is indicated. Metallic-pin fixation of the joint should be avoided, as Steinmann pins, Kirschner wires, threaded pins with bent ends, and Hagie pins have all been reported to migrate and cause serious complications, including death. PMID- 10797195 TI - Transmission of Human Immunodeficiency Virus Infection in the Surgical Setting. AB - The emergence of the human immunodeficiency virus (HIV) has highlighted the need for orthopaedic surgeons to understand the epidemiology of percutaneous injuries and other blood exposures in the surgical setting. The American Academy of Orthopaedic Surgeons and the Centers for Disease Control and Prevention have worked to increase understanding and prevent transmission of blood-borne viral diseases in orthopaedic surgery. This article addresses the risk of HIV transmission in the surgical setting, with a focus on surveillance efforts to monitor the extent of occupational HIV infection, specific risk factors, and postexposure management. Health-care worker-to-patient transmission and patient to-patient transmission are also addressed. PMID- 10797196 TI - Patellar Tendon Ruptures. AB - Rupture of the patellar tendon is a relatively infrequent, yet disabling, injury, which is most commonly seen in patients less than 40 years of age. It tends to occur during athletic activities when a violent contraction of the quadriceps muscle group is resisted by the flexed knee. Rupture usually represents the final stage of a degenerative tendinopathy resulting from repetitive microtrauma to the patellar tendon. This injury may also occur during less strenuous activity in patients whose tendons are weakened by systemic illness or the administration of local or systemic corticosteroid medications. The diagnosis is made on the basis of the presence of a painful, palpable defect in the substance of the tendon; an inability to completely extend the knee against gravity; and the existence of patella alta confirmed by lateral radiographs. Ultrasonography and magnetic resonance imaging are useful in identifying a neglected rupture, as well as when the diagnosis is in question or an intra-articular injury is suspected. The prognosis after a patellar tendon rupture depends in large part on the interval between injury and repair. Surgery soon after the injury is recommended for optimal results. This is best accomplished by accurate reapproximation of the ruptured tendon ends, repair of the torn extensor retinacula, and placement of a reinforcing cerclage suture. An aggressive rehabilitation program, emphasizing early range-of-motion exercises, protected weight bearing, and quadriceps strengthening, will enhance the results of surgery. Patients who undergo delayed repair are at risk for a compromised result secondary to loss of full knee flexion and decreased quadriceps strength, although a functional extensor mechanism is likely to be reestablished. PMID- 10797197 TI - Human Immunodeficiency Virus Infection: Complications and Outcome of Orthopaedic Surgery. AB - Orthopaedic surgeons practicing in areas with a high prevalence of human immunodeficiency virus (HIV) infection may expect that up to 7% of their patients who undergo emergent procedures and 1% to 3% of those who undergo elective surgery will be HIV-positive. Although basic science studies have demonstrated impairment of defenses to routine orthopaedic pathogens as well as to opportunistic organisms, clinical studies have shown that this impairment has not resulted in an increased incidence of postoperative infections or failure of wound healing in the asymptomatic HIV-positive patient. Even for the symptomatic patient, current medical management appears adequate to reduce the risk of early postoperative infection. The HIV-positive patient with a pros-thetic implant may be at increased risk for late hematogenous implant infection as host defenses diminish. Regular medical attention, prophylactic antibiotic therapy before dental work and invasive procedures, and early evaluation and treatment of possible infections are especially important in this setting. Decisions regarding elective surgery should be made on a risk-benefit basis. Because the risk of surgical complications increases with progression of the dis-ease, guidelines for elective surgery should include an assessment of the HIV-positive patient's immune status, including the CD4 lymphocyte count, history of opportunistic infection, serum albumin level, the presence of skin anergy, and the state of nutrition and general health. PMID- 10797198 TI - Cervical Radiculopathy: Diagnosis and Nonoperative Management. AB - Cervical radiculopathy presents as pain in a dermatomal distribution. This frequently represents compression of an exiting cervical nerve root by either a herniated disk or a degenerative cervical spondylotic change. Most patients will improve with nonoperative treatment, and a small percentage will require further diagnostic evaluation and ultimately surgical intervention. An understanding of the normal anatomy and the pathologic changes in cervical radiculopathy will improve the understanding of diagnosis and decision making regarding nonoperative interventions. An algorithmic approach for decision making and a review of nonoperative management are presented. PMID- 10797199 TI - Distraction Histiogenesis: Principles and Indications. AB - Distraction histiogenesis is a biologic phenomenon that can be utilized to induce the formation of new bone and soft tissue. This technique has been used after corticotomy or osteotomy of bone to treat patients with limb-length inequality, angular deformities, segmental bone loss, nonunions, and contractures. A distraction force is applied with an external fixator, such as the Ilizarov circular fixator or a uniplanar fixator. The authors review the extensive preoperative planning required, the performance of osteotomy, the application of external fixators, and the timing between the osteotomy and the initiation of correction (the latency phase). The subsequent distraction phase involves active lengthening, transport, or angular correction through frequent small steps (e.g., 0.25 mm every 6 hours). This results in the formation of new bone, or regenerate, in longitudinal columns along the plane of distraction. The consolidation phase begins after the desired correction has been achieved; this period allows for maturation of the regenerate and corticalization before fixator removal. PMID- 10797200 TI - Interdigital Neuritis: Diagnosis and Treatment. AB - Because it has not yet been established whether the condition commonly referred to as Morton's neuroma results from true neuromatous proliferation or from inflammation in the region of the interdigital nerve, the term "interdigital neuritis" is preferred. The authors review the etiology, diagnosis, and management of interdigital neuritis, including whether a plantar or dorsal approach is preferable and whether neurectomy is more efficacious than incision of the transverse metatarsal ligament, with or without neurolysis. The authors recommend that diagnosis be made on the basis of the history and clinical examination, that surgery be performed through a dorsal approach with release of the transverse ligament but without neurectomy, and that revision surgery be performed through a dorsal incision with excision of the nerve 3 cm proximal to the transverse ligament. PMID- 10797201 TI - Distal Humeral Fractures in Adults. AB - Distal humeral fractures in adults often pose a challenge to the orthopaedic surgeon. Preoperative planning, minimal devitalization of bone and soft tissue, and adherence to the prerequisites of biomechanical fixation are all important elements in effecting the desired end result. The chevron modification of the olecranon osteotomy affords excellent surgical exposure of the joint surface for fractures with an intra-articular component. When two plates are used to fix the lateral and medial distal humeral columns, it is best to orient them so that, when looked at in cross section, they are at right angles to each other. The achievement and maintenance of an anatomic reduction secure enough to permit early functional, pain-free motion of the elbow can be best ensured by open reduction and internal fixation with careful attention to detail. PMID- 10797202 TI - Displaced Fractures of the Radial Head: Internal Fixation or Excision? AB - Displaced fractures of the radial head in the young active patient should no longer be routinely treated with excision of the radial head. Better techniques of imaging, surgical exposure, and implant placement have improved the likelihood of preserving the head. Associated injuries may make preservation of the radial head important for both acute and long-term stability. In patients with suspected injury to the interosseous ligament of the forearm, saving the radial head may prevent pathologic proximal migration. Rigid internal fixation, permitting early mobilization, can be applied to the radial head and neck in a "safe zone" that does not impede motion. Radial-head excision should be performed in patients with grossly comminuted fractures and in those with low demand on their upper extremities. PMID- 10797203 TI - Acromioclavicular Joint Injuries and Distal Clavicle Fractures. AB - The acromioclavicular joint is commonly affected by traumatic and degenerative conditions. Most injuries are due to direct trauma, such as a fall on the shoulder. Six types of acromioclavicular sprains and three types of distal clavicle fractures have been described in adults. Although there is general agreement on treatment of type I, II, IV, V, and VI acromioclavicular injuries, the treatment of type III injuries remains controversial. Studies have shown no distinct advantage for surgical reconstruction over nonoperative treatment. Because type II distal clavicle fractures are prone to nonunion, operative fixation may be advisable to avoid this complication. PMID- 10797204 TI - Supracondylar Fractures of the Humerus in Children. AB - The treatment of type II and type III supracondylar fractures of the humerus in children with closed reduction and percutaneous pinning has dramatically lowered the rate of complications from this injury. The incidence rates of malunion (cubitus varus) and compartment syndrome have both decreased. Nerve injury accompanying this type of fracture (prevalence, 5% to 19%) is usually a neurapraxia, which should be managed conservatively. Vascular insufficiency at presentation (prevalence, 5% to 17%) should be managed initially by rapid closed reduction and pinning without arteriography. Persistent vascular insufficiency necessitates exploration and vascular reconstruction. PMID- 10797205 TI - Hip Dislocation: Current Treatment Regimens. AB - Dislocation of the hip occurs only with high-energy trauma, and concomitant injuries are common. Early diagnosis and institution of treatment are necessary to obtain the best possible results. Treatment protocols include emergent reduction of the femoral head to reestablish perfusion, postreduction radiography and computed tomography to look for associated fractures and to judge the concentricity of the reduction, stability testing, and early mobilization. Open reduction may be required if a concentric reduction cannot be obtained in a closed manner. Despite appropriate management, posttraumatic arthritis and avascular necrosis may occur, with reported rates as high as 15% to 30%. Patients who sustain a hip dislocation should be made aware of these potential complications at the time of initial treatment. PMID- 10797206 TI - Soft-Tissue Injuries Associated With High-Energy Extremity Trauma: Principles of Management. AB - The management of high-energy extremity trauma has evolved over the past several decades, and appropriate treatment of associated soft-tissue injuries has proved to be an important factor in achieving a satisfactory outcome. Early evaluation of the severely injured extremity is crucial. Severe closed injuries require serial observation of the soft tissues and early skeletal stabilization. Open injuries require early aggressive debridement of the soft tissues followed by skeletal stabilization. Temporary wound dressings should remain in place until definitive soft-tissue coverage has been obtained. Definitive soft-tissue closure will be expedited by serial debridements performed every 48 to 72 hours in a sterile environment. Skeletal union is facilitated by early bone grafting and/or modification of the stabilizing device. Aggressive rehabilitation, includ-ing early social reintegration, are crucial for a good functional outcome. Adherence to protocols is especially beneficial in the management of salvageable severely injured extremities. PMID- 10797207 TI - Patellofemoral Instability: Evaluation and Management. AB - Patellofemoral disorders are a common cause of knee pain and disability. A thorough history and a careful physical examination are essential to accurate diagnosis, and imaging modalities play an important role. Magnetic resonance imaging can provide information on malalignment and soft-tissue injuries. Although there is a continuum of diagnoses, most patellofemoral disorders can be divided into three distinct categories: soft-tissue abnormalities, patellar instability due to subluxation and dislocation, and patellofemoral arthritis. Many patellofemoral disorders respond to nonoperative therapy. When surgical intervention is necessary, patellar tilt can be successfully treated by a lateral release. Lateral patellar subluxation associated with malalignment can be corrected by a distal realignment procedure such as the anteromedial tibial tubercle transfer. Repair of the medial patellofemoral ligament in cases of patellar dislocation has considerably lowered the incidence of recurrent instability. Although no ideal treatment exists for patellofemoral arthritis, mechanical symptoms may be alleviated by arthroscopic debridement of delamination lesions. Articular cartilage-wear disorders may be stabilized by addressing the primary causative disorder. PMID- 10797208 TI - Extensor Tendon Injuries in the Hand. AB - Until recently, extensor tendon injuries were often discounted as an important hand problem. However, studies have shown that not all extensor lacerations fare well and that loss of flexion can be problematic. Newer postoperative protocols emphasizing tendon gliding have improved results, and better repair techniques and postoperative rehabilitation regimens are under investigation. This article reviews the evaluation of acute open and closed extensor tendon injuries, their conservative and surgical treatment, and postoperative rehabilitation options. PMID- 10797209 TI - Back Pain in Children and Adolescents: Evaluation and Differential Diagnosis. AB - Back pain in children and adolescents usually has a recognizable organic origin. The most common entities seen are spondylolysis, spondylolisthesis, Scheuermann's kyphosis, disk herniations, infections, and tumors. Early recognition and treatment can provide patients the best chance at relief of symptoms and eradication of the underlying disease process. The goals of this review are to (1) familiarize the clinician with the various diagnoses associated with back pain in the skeletally immature patient and (2) to assist the clinician in making the appropriate diagnosis by providing a rational method of selecting diagnostic tests that maximize specificity and minimize costs. PMID- 10797210 TI - Metastatic Disease of the Hip: Evaluation and Treatment. AB - Lesions in the area of the hip secondary to metastatic disease present challenging problems for the orthopaedic surgeon. With the advent of improved medical therapies for many types of cancer have come not only an increase in life expectancy but also an increased likelihood that symptomatic metastatic bone lesions will appear. Advances in internal fixation have enabled the orthopaedic surgeon to provide an increased level of comfort and mobility to many patients with metastatic disease. PMID- 10797211 TI - The Female Athlete: Evaluation and Treatment of Sports-Related Problems. AB - Although many of the problems faced by the female athlete affect the male athlete as well, some occur exclusively or more commonly in women. These include spondylolisthesis, stress fractures in the pelvis and hip, and pelvic floor dysfunction. Female athletes are also more likely to have patellofemoral problems, noncontact anterior cruciate ligament tears, and bunions. For many of these conditions, the relative influences of osseous anatomy, ligamentous laxity, and the effect of sex hormones have not yet been established. There are also problems related specifically to the menstrual cycle and pregnancy. Amenorrhea is present in up to 20% of vigorously exercising women. The term "female athlete triad" has been coined to describe the complex interplay of menstrual irregularity, disordered eating, and premature osteoporosis seen in the female athlete. Many of the concerns related to exercise during pregnancy focus on the safety of the fetus rather than the athlete herself. Musculoskeletal problems in the physically active pregnant woman are related to weight gain, ligamentous relaxation, lordosis, and change in the center of gravity. PMID- 10797212 TI - Painful Shoulder After Surgery for Rotator Cuff Disease. AB - Persistent shoulder pain after surgery for rotator cuff disease may be caused by conditions that are either extrinsic or intrinsic to the shoulder. Extrinsic causes of persistent shoulder pain include cervical radiculopathy, suprascapular neuropathy, abnormalities of scapular rotation (due to long-thoracic or spinal accessory neuropathy), and adjacent or metastatic neoplasms. Causes of persistent pain that are intrinsic to the shoulder include both intra-articular conditions (e.g., glenohumeral osteoarthritis, adhesive capsulitis, recurrent anterior subluxation, and labral and bicipital tendon abnormalities) and extra-articular conditions (e.g., persistent subacromial impingement, persistent or recurrent rotator cuff defects, acromioclavicular arthropathy, and deltoid muscle deficiency). Successful management requires an accurate diagnosis, maximal rehabilitation, judicious use of surgical intervention, and a well-motivated patient. The results of revision surgery in patients with persistent subacromial impingement, with or without an intact cuff, are inferior to reported results after primary acromioplasty or rotator cuff repair. PMID- 10797213 TI - Plantar Fasciitis: Diagnosis and Conservative Management. AB - Plantar fasciitis is a common cause of heel pain, which frustrates patients and practitioners alike because of its resistance to treatment. It has been associated with obesity, middle age, and biomechanical abnormalities in the foot, such as tight Achilles tendon, pes cavus, and pes planus. It is considered to be most often the result of a degenerative process at the origin of the plantar fascia at the calcaneus. However, neurogenic and other causes of subcalcaneal pain are frequently cited. A combination of causative factors may be present, or the true cause may remain obscure. Although normally managed with conservative treatment, plantar fasciitis is frequently resistant to the wide variety of treatments commonly used, such as nonsteroidal anti-inflammatory drugs, rest, pads, cups, splints, orthotics, corticosteroid injections, casts, physical therapy, ice, and heat. Although there is no consensus on the efficacy of any particular conservative treatment regimen, there is agreement that nonsurgical treatment is ultimately effective in approximately 90% of patients. Since the natural history of plantar fasciitis has not been established, it is unclear how much of symptom resolution is in fact due to the wide variety of commonly used treatments. PMID- 10797214 TI - Complex Articular Fractures of the Distal Radius: Classification and Management. AB - Articular fractures of the distal radius require an anatomic reduction, as even minimal step-offs are associated with the development of osteoarthritis. Such fractures are classified on the basis of both the mechanism and the pattern of injury. The comprehensive classification of fractures established by Muller et al defines those occurring from a shearing mechanism and involving part of the articular surface as type B fractures. These are in turn subdivided into group B1 fractures, which involve the radial styloid (chauffeur's fracture); group B2, which involve the dorsal margin of the radius; and group B3, which involve the volar margin of the distal radius (Barton's fracture). Type B fractures are unstable and often require operative intervention. Articular fractures resulting from a compression force on the end of the radius are classified as type C. A group C1 fracture is a two-part fracture without metaphyseal comminution; group C2, a two-part fracture with metaphyseal comminution; group C3, a fracture with more than two pieces, with or without comminution. The operative tactic for compression fractures involves restoration of the four common fragments in sequence. This can often be accomplished by manipulative means alone or with limited exposure of the fracture fragments and stabilization with percutaneous Kirschner wires. Neutralization with an external fixator and use of an autogenous bone graft are often required. PMID- 10797215 TI - Frozen Shoulder: Diagnosis and Management. AB - "Frozen shoulder" comprises a group of conditions caused by different processes. Effective treatment depends on recognition of the underlying pathologic dis-order in each individual case. Idiopathic adhesive capsulitis usually responds to nonoperative therapy or closed manipulation, but shoulder stiffness due to trauma or surgery may necessitate either an arthroscopic or an open-release procedure. Both of these technically demanding techniques are effective in restoring motion in cases of frozen shoulder refractory to nonoperative treatment. PMID- 10797216 TI - Soft-Tissue Tumors About the Knee. AB - Soft-tissue tumors about the knee include a wide variety of entities, ranging from synovial cysts to aggressive high-grade sarcomas. The overlap in clinical presentation of these various masses frequently results in misdiagnosis and inappropriate treatment. Unnecessary and sometimes costly arthroscopy sometimes precedes the diagnosis of soft-tissue sarcoma about the knee. A poorly planned or executed biopsy has been demonstrated to have an adverse effect on patient prognosis and may lead to unnecessary amputation. Special vigilance in evaluation is warranted when a soft-tissue mass is not in the typical position or does not have other characteristic features of a meniscal or Baker's cyst, when the size of the mass or the accompanying symptoms seem out of proportion to the injury or underlying degenerative process, and when symptoms persist beyond what is expected. When malignancy is suspected, the patient should be referred to a musculoskeletal oncologist before biopsy. PMID- 10797217 TI - Pseudarthrosis of the Lumbar Spine. AB - Pseudarthrosis can be a costly and disabling complication of lumbar spinal fusion. This review focuses on the incidence, causation, diagnosis, and nonoperative management of this condition, as well as surgical approaches that can be effective in treating carefully selected patients. Judicious initial selection of patients for fusion and the use of meticulous surgical technique in the first operation continue to be the best means of prevention. PMID- 10797218 TI - Supracondylar Fractures of the Femur. AB - Successful management of the distal femoral fracture is possible with adherence to the basic principles of anatomic reduction, stable fixation, and early motion. Closed management can achieve these goals in selected patients, but most supracondylar femoral fractures are better treated with operative reconstruction. Implant selection is determined on the basis of the characteristics of the fracture, the bone quality, the needs of the patient, and the experience of the surgeon. Surgical options include the angled blade plate, compression-screw systems, condylar buttress plates, intramedullary nails, external fixation, and modular distal femoral replacement. The author reviews the indications and techniques for using these devices. PMID- 10797219 TI - Injuries to the Distal Lower Extremity Syndesmosis. AB - Disruption of the distal syndesmosis of the lower extremity is most commonly associated with ankle fractures but can also occur without gross bone injury. Definitive management of these injuries remains controversial. The current indications for syndesmosis fixation are based on tibiotalar joint mechanics as determined in cadaveric and biomechanical studies, as well as radiologic evaluation and an understanding of the pertinent anatomy and the etiology of these injuries. Such data support the use of syndesmotic screws in selected fractures that include a disruption of the syndesmosis. However, definitive fixation recommendations for syndesmosis disruption with or without ankle fracture remain under investigation. Distal lower extremity syndesmosis sprains without fracture or subluxation consistently require longer recovery time than typical lateral sprains and can be associated with greater long-term disability. PMID- 10797220 TI - Calcific Tendinopathy of the Rotator Cuff: Pathogenesis, Diagnosis, and Management. AB - Calcific tendinopathy, or calcifying tendinitis, is a disease characterized by multifocal, cell-mediated calcification of living tissue. After spontaneous disappearance of the calcific deposits or, less frequently, surgical removal, the tendon reconstitutes itself. Attention to the clinical presentation and the radiologic, morphologic, and gross characteristics of the calcium deposit will facilitate differentiation between the formative phase and the resorptive phase, which is of paramount importance in the management of this disease. Should conservative treatment fail, surgical removal may be indicated during the formative phase, but only under exceptional circumstances during the resorptive phase. Aspiration and lavage of the deposit should be performed only during the latter phase. PMID- 10797221 TI - Hindfoot Dislocations: When Are They Not Benign? AB - Acute hindfoot dislocations are usually characterized by displacement of both the talocalcaneal and the talonavicular joints. Medial dislocations are more common than lateral ones. Closed reduction is usually obtained easily. When closed attempts fail, surgical exploration and removal of recognized obstacles to reduction are necessary. Associated open wounds necessitate aggressive operative management to prevent infection. Postreduction radiographs should be scrutinized for the presence of associated fractures that require fixation or surgical removal. A short-leg walking cast should be used for 3 to 6 weeks. In rare instances, the tibiotalar joint is also dislocated, which usually necessitates open reduction or, if the injury is open, extruded, and contaminated, talar excision. All hindfoot dislocations result in some stiffening of the hindfoot. Painful degenerative arthrosis sometimes develops after this injury. Factors that predispose to poor outcomes include high-energy mechanisms, the presence of open wounds and fractures, and lateral dislocations. Painful arthrosis that does not respond to conservative treatment can be treated with selective hindfoot arthrodesis. PMID- 10797222 TI - Odontoid Fractures: Evaluation and Management. AB - Fractures of the odontoid process are uncommon injuries. Fracture displacement, compromised blood supply, comminution, and iatrogenic distraction have all been implicated in the reported high rates of nonunion. Plain radiography, polytomography, and computed tomography are all useful in delineating the fracture pattern. Magnetic resonance imaging has been recommended for evaluating associated ligamentous injuries and may be helpful in detecting occult cervical spine fractures. Type I fractures are avulsion fractures of the tip of the odontoid process. These rare injuries require only external immobilization with an orthosis if there is no associated ligamentous injury. Type II fractures occur at the junction of the odontoid process and the body of the axis. These are the most common odontoid fractures and are associated with a high incidence of nonunion. Nondisplaced fractures should be treated with halo immobilization for 8 to 12 weeks, with careful clinical and radiographic monitoring. Displaced fractures should be considered for operative treatment, either with atlantoaxial arthrodesis or anterior screw fixation. Type III fractures, which extend into the body of the axis through cancellous bone, are treated with closed reduction and halo immobilization. PMID- 10797223 TI - Obstetric Brachial Plexus Injuries: Evaluation and Management. AB - Most infants with brachial plexus birth palsy who show signs of recovery in the first 2 months of life will subsequently have normal function. However, infants who do not recover in the first 3 months of life have a considerable risk of long term limited strength and range of motion. As the delay in recovery extends from 3 months to beyond 6 months, this risk increases pro-portionately. The presence of a total plexus lesion, a partial plexus lesion with loss at C5-C7, or Horner's syndrome carries a worse prognosis. Microsurgery is indicated for failure of return of function by 3 to 6 months. The exact timing of intervention is still open to debate. With microsurgical reconstruction, there is improvement in outcome in a high percentage of patients. However, the neural lesion is too severe and complex for present methods of reconstruction to restore normal function. Secondary correction of shoulder dysfunction with either latissimus dorsiteres major tendon transfer or humeral derotation osteotomy is clearly beneficial for patients with chronic brachial plexopathy, as is reconstruction of supination forearm contracture with biceps rerouting transfer and/or forearm osteotomy. Reconstruction of the hand is also indicated for the patient with chronic disability. All of these procedures improve, but do not completely normalize, function. PMID- 10797224 TI - Compression Plating Versus Intramedullary Fixation of Humeral Shaft Fractures. AB - Most humeral shaft fractures do not require surgery. When operative stabilization is indicated, the surgeon can choose between compression plating and intramedullary fixation. The results after compression plating have been shown to be predictable with respect to healing, alignment, and range of motion of the shoulder and elbow joints. Although complications are unusual with plate fixation, the procedure can require extensive dissection and operative time. Intramedullary fixation offers an alternative to plate fixation, with the principal advantage being a limited surgical dissection. This benefit must be balanced against the reportedly high rate of postoperative shoulder problems seen with antegrade nail placement. Unfortunately, few direct comparative studies have been done to evaluate the various techniques. The authors attempt to clarify and resolve these issues. PMID- 10797225 TI - Injuries to the Ulnar Collateral Ligament of the Thumb Metacarpophalangeal Joint. AB - Injuries to the ulnar collateral ligament of the metacarpophalangeal (MCP) joint of the thumb are relatively common. When an incomplete rupture is present, valgus stress testing with the MCP joint positioned in extension reveals minimal or no instability (less than 30 degrees of laxity or less than 15 degrees more laxity than in the noninjured thumb). When a complete rupture is present, valgus stress testing with the MCP joint positioned in extension reveals marked laxity (more than 30 degrees or more than 15 degrees more laxity than in the noninjured thumb). In this instance, displacement of the ligament proximal and superficial to the adductor aponeurosis, which is often termed a Stener lesion, is likely. Partial ligament injuries in which the ligament is not displaced may be treated nonoperatively. When a Stener lesion is present, however, primary ligament healing cannot occur without operative management. Whether treatment should be operative or nonperative can generally be decided on the basis of the findings from the history, the radiographs, and the physical examination, which should include valgus stress testing. PMID- 10797227 TI - Editorial: peptide and peptidomimetic ligands of opioid receptors PMID- 10797226 TI - Elbow Fractures in Children: Diagnosis and Management. AB - Fractures about the elbow are very common in children, but the anatomy of the child's elbow may make the diagnosis less obvious than in a mature skeleton. An understanding of the ossification and fusion of the secondary growth centers about the elbow is essential to avoid overlooking these injuries and to optimize treatment. If plain radiographs are equivocal, an arthrogram of the elbow may clarify the anatomy and diagnosis. Early neurologic and vascular complications are not uncommon and must be recognized before fracture reduction. Many late complications, such as malunion, osteonecrosis, and physeal bridging, are largely preventable by correct early diagnosis and treatment. Anatomic reduction of articular surfaces, restoration of physeal anatomy, and near-anatomic alignment of fracture fragments in the frontal plane are the corner-stones of successful treatment of pediatric elbow fractures. PMID- 10797228 TI - New opioid peptides, peptidomimetics, and heterocyclic compounds from combinatorial libraries. AB - Here we review the use of combinatorial libraries in opioid receptor assays. Following a brief description of the history of the combinatorial field, methods for the generation of synthetic libraries and the deconvolution of mixture-based libraries are presented. Case studies involving opioid assays used to demonstrate the viability of combinatorial libraries are described. The identification of new opioid peptides from combinatorial libraries is reviewed. The peptides found are composed of L-amino acids, D-amino acids, or L-, D-, and unnatural amino acids, and range from tetrapeptides to decapeptides. Likewise, new opioid compounds identified from peptidomimetic libraries, such as peptoids and alkylated dipeptides, and those identified from acyclic (e.g., polyamine, urea) and heterocyclic (e.g., bicyclic guanidine) libraries, are reviewed. PMID- 10797229 TI - Conformation-activity relationships of opioid peptides with selective activities at opioid receptors. AB - The discovery of endogenous opioid peptides 25 years ago opened up a new chapter in efforts to understand the origins and control of pain, its relationships to other biological functions, including inflammatory and other immune responses, and the relationships of opioid peptides and their receptors to a variety of undesirable or toxic side effects often associated with the nonpeptide opiates such as morphine including addiction, constipation, a variety of neural toxicities, tolerance, and respiratory depression. For these investigations the need for potent and highly receptor selective agonists and antagonists has been crucial since they in principle allow one to distinguish unequivocally the roles of the different opioid receptors (mu, delta, and kappa) in the various biological and pathological roles of the opioid peptides and their receptors. Conformational and topographical constraint of the linear natural endogenous opioid peptides has played a major role in developing peptide ligands with high selectivity for mu, delta, and kappa receptors, and in understanding the conformational, topographical, and stereoelectronic structural requirements of the opioid peptides for their interactions with opioid receptors. In turn, this had led to insights into the three-dimensional pharmacophore for opioid receptors. In this article we review and discuss some of the developments that have led to potent, selective, and stable peptide and peptidomimetic ligands that are highly potent and selective, and that have delta agonist, mu antagonist, and kappa agonist biological activities (other authors in this issue will discuss the development of other types of activities and selectivities). These have led to ligands that provide unique insight into opioid pharmacophores and the critical roles opioid ligands and receptor scan play in pain, addiction, and other human maladies. PMID- 10797230 TI - The TIPP opioid peptide family: development of delta antagonists, delta agonists, and mixed mu agonist/delta antagonists. AB - The discovery of the prototype delta opioid antagonists TIPP (H-Tyr-Tic-Phe-Phe OH) and TIP (H-Tyr-Tic-Phe-OH) in 1992 was followed by extensive structure activity relationship studies, leading to the development of analogues that are of interest as pharmacological tools or as potential therapeutic agents. Stable TIPP-derived delta opioid antagonists with subnanomolar delta receptor binding affinity and extraordinary delta receptor selectivity include TIPP[Psi] (H-Tyr TicPsi[CH(2)NH]Phe-Phe-OH] and TICP[Psi] (H-Tyr-TicPsi[CH(2)NH]Cha-Phe-OH); Cha: cyclohexylalanine), which are widely used in opioid research. Theoretical conformational analyses in conjunction with the pharmacological characterization of conformationally constrained TIPP analogues led to a definitive model of the receptor-bound conformation of H-Tyr-Tic-(Phe-Phe)-OH-related delta opioid antagonists, which is characterized by all-trans peptide bonds. Further structure activity studies revealed that the delta antagonist vs delta agonist behavior of TIP(P)-derived compounds depended on very subtle structural differences in diverse locations of the molecule and suggested a delta receptor model involving a number of different inactive receptor conformations. A further outcome of these studies was the identification of a new class of potent and very selective dipeptide delta agonists of the general formula H-Tyr-Tic-NH-X (X = arylalkyl), which are of interest for drug development because of their low molecular weight and lipophilic character. Most interestingly, TIPP analogues containing a C terminal carboxamide group displayed a mixed mu agonist/delta antagonist profile, and thus were expected to be analgesics with a low propensity to produce tolerance and physical dependence. This turned out to be the case with the TIPP derived mu agonist/delta antagonist DIPP-NH(2)[Psi] (H-Dmt-TicPsi[CH(2)NH]Phe-Phe NH(2)); Dmt: 2',6'- dimethyltyrosine). PMID- 10797231 TI - Complementarity of delta opioid ligand pharmacophore and receptor models. AB - The elaboration of a pharmacophore model for the delta opioid receptor selective ligand JOM-13 (Tyr-c[D-Cys-Phe-D-Pen]OH) and the parallel, independent development of a structural model of the delta receptor are summarized. Although the backbone conformation of JOM-13's tripeptide cycle is well defined, considerable conformational lability is evident in the Tyr(1) residue and in the Phe(3) side chain, key pharmacophore elements of the ligand. Replacement of these flexible features of the ligand by more conformationally restricted analogues and subsequent correlation of receptor binding and conformational properties allowed the number of possible binding conformations of JOM-13 to be reduced to two. Of these, one was chosen as more likely, based on its better superposition with other conformationally constrained delta receptor ligands. Our model of the delta opioid receptor, constructed using a general approach that we have developed for all rhodopsin-like G protein-coupled receptors, contains a large cavity within the transmembrane domain that displays excellent complementarity in both shape and polarity to JOM-13 and other delta ligands. This binding pocket, however, cannot accommodate the conformer of JOM-13 preferred from analysis of ligands, alone. Rather, only the "alternate" allowed conformer, identified from analysis of the ligands but "disfavored" because it does not permit simultaneous superposition of all pharmacophore elements of JOM-13 with other delta ligands, fits the binding site. These results argue against a simple view of a single, common fit to a receptor binding site and suggest, instead, that at least some binding site interactions of different ligands may differ. PMID- 10797232 TI - Mutational analysis of the structure and function of opioid receptors. AB - The cloning of the opioid receptors allows the investigation of receptor domains involved in the peptidic and nonpeptidic ligand interaction and activation of the opioid receptors. Receptor chimera studies and mutational analysis of the primary sequences of the opioid receptors have provided insights into the structural domains required for the ligand recognition and receptor activation. In the current review, we examine the current reports on the possible involvement of extracellular domains and transmembrane domains in the high-affinity binding of peptidic and nonpeptidic ligands to the opioid receptor. The structural requirement for the receptors' selectivity toward different ligands is discussed. The receptor domains involved in the activation and subsequent cellular regulation of the receptors' activities as determined by mutational analysis will also be discussed. Finally, the validity of the conclusions based on single amino acid mutations is examined. PMID- 10797234 TI - Preparation, morphological characterization, and activity of thin films of horseradish peroxidase. AB - Active uniform films of horseradish peroxidase (HRP) have been prepared by covalent binding on Si/SiO(2) or glass supports previously activated by silanization and succinylation. Labeling by fluorescent or by Electron Spin Resonance (ESR) probes was used to quantify the surface density of active groups and of horseradish peroxidase. Atomic Force Microscopy (AFM) imaging was used to characterize the surface morphology. We observed that a non-uniform protein adsorption due to physical interactions was present when the supports were not activated for covalent binding and was, in large part, removed by washing. The enzyme deposited by covalent binding formed homogeneous layers with a height in the range 60-90 A. By using a fluorescent label, we calculated a protein density of 3.6 x 10(12) molecules cm(-2) on Si/SiO(2), corresponding to an estimated area per molecule of 2800 A(2) which is in agreement with the value expected on the basis of the crystallographic data considering the formation of a monomolecular layer. The protein density of the layer immobilized on glass was similar (1.9 x10(12) molecules cm(-2)). The enzyme immobilized on both supports showed a k(cat)/K(M) being of the order of 3-5x10(5) M(-1)s(-1) that is 1/20th of free HRP. The half-life time of the activity of the enzyme immobilized by covalent binding was longer than 40 days at 6 degrees C. PMID- 10797233 TI - ATR-FTIR sensor development for continuous on-line monitoring of chlorinated aliphatic hydrocarbons in a fixed-bed bioreactor. AB - This article describes the continuous on-line monitoring of a dechlorination process by a novel attenuated total reflection-Fourier transform infrared (ATR FTIR) sensor. This optical sensor was developed to measure noninvasively part-per million (ppm) concentrations of trichloroethylene (TCE), tetrachloroethylene (PCE), and carbon tetrachloride (CT) in the aqueous effluent of a fixed-bed dechlorinating bioreactor, without any prior sample preparation. The sensor was based on an ATR internal reflection element (IRE) coated with an extracting hydrophobic polymer, which prevented water molecules from interacting with the infrared (IR) radiation. The selective diffusion of chlorinated compound molecules from aqueous solution into the polymer made possible their detection by the IR beam. With the exclusion of water the detection limits were lowered, and measurements in the low ppm level became possible. The best extracting polymer was polyisobutylene (PIB) in the form of a 5.8-microm thick film, which afforded a detection limit of 2, 3, and 2. 5 mg/L (ppm) for TCE, PCE, and CT, respectively. Values of the enrichment factors between the polymer coating and the water matrix of these chloro-organics were determined experimentally and were compared individually with predictions obtained from the slopes of absorbance/concentration curves for the three analytes. Before coupling the ATR FTIR sensor to the dechlorinating bioreactor, preliminary spectra of the chlorinated compounds were acquired on a laboratory scale configuration in stop flow and flow-through closed-loop modes. In this way, it was possible to study the behavior and direct response of the optical sensor to any arbitrary concentration change of the analytes. Subsequently, the bioreactor was monitored with the infrared sensor coupled permanently to it. The sensor tracked the progression of the analytes' spectra over time without perturbing the dechlorinating process. To calibrate the ATR-FTIR sensor, a total of 13 standard mixtures of TCE, PCE and CT at concentrations ranging from 0 to 60 ppm were selected according to a closed symmetrical experimental design derived from a 3(2) full-factorial design. The above range of concentrations chosen for calibration reflected typical values during normal bioreactor operation. Several partial least squares (PLS) calibration models were generated to resolve overlapping absorption bands. The standard error of prediction (SEP) ranged between 0.6 and 1 ppm, with a relative standard error of prediction (RSEP) between 3 and 6% for the three analytes. The accuracy of this ATR-FTIR sensor was checked against gas chromatography (GC) measurements of the chlorocompounds in the bioreactor effluents. The results demonstrate the efficiency of this new sensor for routine continuous on-line monitoring of the dechlorinating bioreactor. This strategy is promising for bioprocess control and optimization. PMID- 10797235 TI - Stoichiometry and kinetics of poly-beta-hydroxybutyrate metabolism under denitrifying conditions in activated sludge cultures. AB - The objective of this research was to obtain a more detailed insight in the kinetics and stoichiometry of poly-beta-hydroxybutyrate (PHB) metabolism in activated sludge cultures. The process of storage and degradation of PHB under denitrifying conditions was studied. Dynamic conditions as occurring in activated sludge processes were simulated in a 2L sequencing batch reactor (SBR) by subjecting a mixed sludge population at a sludge retention time (SRT) of about 6 days to successive periods of external substrate availability (feast period) and no external substrate availability (famine period). Under these conditions intracellular storage and degradation of PHB occurs. It was shown that the process of storage and degradation of PHB is the same under anoxic and aerobic conditions. About 70% of the amount of acetate consumed in the feast period is used for synthesis of PHB, and the remainder is used for growth processes. The anoxic specific acetate uptake rate was 3-4 times lower than aerobically. The PHB metabolism was evaluated on the basis of a metabolic model. In a traditional macroscopic model, 10 parameters would be needed to describe the process. By relating these parameters to metabolic coefficients, only 2 parameters were needed in the description. Degradation of PHB could be described by a power law equation in PHB concentration. The order n in these experiments equaled 0.59, close to a value previously reported for PHB degradation by bio-P-cultures. However, PHB degradation by bio-P-cultures occurs at a much higher rate. Storage of PHB in the feast period can increase the required COD/N ratio for denitrification by 70% compared to a situation without storage. PMID- 10797236 TI - Effect of vapor-phase bioreactor operation on biomass accumulation, distribution, and activity: linking biofilm properties to bioreactor performance. AB - Excess biomass accumulation and activity loss in vapor-phase bioreactors (VPBs) can lead to unreliable long-term operation. In this study, temporal and spatial variations in biomass accumulation, distribution and activity in VPBs treating toluene-contaminated air were monitored over a 96-day period. Two laboratory scale bioreactors were subjected to a toluene loading rate of 45.8 g/m(3)-h with one VPB operating in a unidirectional (UD) mode and a second identical VPB operating in a directionally switching (DS) mode. In the UD bioreactor, the contaminated air stream was continuously fed to the bottom of the reactor, while, in the DS bioreactor, the direction of the contaminated gas flow was reversed every three days. Overall, the DS system performed better with respect to biomass distribution and microbial activity across the bioreactor, resulting in more stable bioreactor performance. In contrast, most of the biomass accumulation and activity was confined to the front half of the UD bioreactor column which caused high pressure drops, rapid activity loss and eventually toluene breakthrough. A carbon balance reveals that excess biomass accumulated continuously in both bioreactors, and biomass yield coefficients were very similar (0.59 g dry biomass/g toluene for the UD and 0.63 g dry biomass/g toluene for the DS). The viable biomass population remained relatively constant in both bioreactors over the operational period, while the inactive biomass fraction steadily increased over the same time frame. Biodegradation activity determined by the dehydrogenase enzyme activity assay was found to be a function of biomass accumulation and reflected pollutant removal profiles along the columns. In addition, biomass activity correlated well with the toluene-degrading fraction of the total bacterial population. PMID- 10797237 TI - Fermentative capacity in high-cell-density fed-batch cultures of baker's yeast. AB - High-cell-density fed-batch processes for bakers' yeast production will involve a low-average-specific growth rate due to the limited oxygen-transfer capacity of industrial bioreactors. The relationship between specific growth rate and fermentative capacity was investigated in aerobic, sucrose-limited fed-batch cultures of an industrial bakers' yeast strain. Using a defined mineral medium, biomass concentrations of 130 g dry weight/L were reproducibly attained. After an initial exponential-feed phase (mu = 0.18 h(-1)), oxygen-transfer limitation necessitated a gradual decrease of the specific growth rate to ca. 0.01 h(-1). Throughout fed-batch cultivation, sugar metabolism was fully respiratory, with a biomass yield of 0.5 g biomass/g sucrose(-1). Fermentative capacity (assayed off line as ethanol production rate under anaerobic conditions with excess glucose) showed a strong positive correlation with specific growth rate. The fermentative capacity observed at the end of the process (mu = 0.01 h(-1)) was only half that observed during the exponential-feed phase (mu = 0.18 h(-1)). During fed-batch cultivation, activities of glycolytic enzymes, pyruvate decarboxylase and alcohol dehydrogenase in cell extracts did not exhibit marked changes. This suggests that changes of fermentative capacity during fed-batch cultivation were not primarily caused by regulation of the synthesis of glycolytic enzymes. PMID- 10797238 TI - Effect of alcohol compounds found in hemicellulose hydrolysate on the growth and fermentation of ethanologenic Escherichia coli. AB - Lignocellulose can be readily hydrolyzed into a mixture of sugars using dilute mineral acids. During hydrolysis, a variety of inhibitors are also produced which include aromatic alcohols from lignin and furfuryl alcohol from pentose destruction. Seven compounds were investigated individually and in binary combinations (catechol, coniferyl alcohol, furfuryl alcohol, guaiacol, hydroquinone, methylcatechol, and vanillyl alcohol). Aromatic alcohols and furfuryl alcohol inhibited ethanol production from xylose in batch fermentations primarily by inhibiting the growth of Escherichia coli LY01, the biocatalyst. The toxicities of these compounds were directly related to their hydrophobicity. Methylcatechol was the most toxic compound tested (MIC = 1.5 g/L). In binary combination, the extent of growth inhibition was roughly additive for most compounds tested. However, combinations with furfuryl alcohol and furfural (furaldehyde) appear synergistic in toxicity. When compared individually, alcohol components which are formed during hemicellulose hydrolysis are less toxic for growth than the aldehydes and organic acids either on a weight basis or a molar basis. PMID- 10797239 TI - Isolation of thermotolerant ethanologenic yeasts and use of selected strains in industrial scale fermentation in an Egyptian distillery. AB - An enrichment and isolation program for new ethanol-producing thermotolerant yeasts as well as a screening program of some known thermotolerant strains resulted in the selection of several strains capable of growth at 40-43 degrees C. Among these strains four grew and fermented sugar cane molasses at 43 degrees C under batch conditions with sugar-conversion efficiencies >94% and ethanol concentrations 6.8-8.0% (w/v). The two best-performing strains, a Saccharomyces cerevisiae F111 and a Kluyveromyces marxianus WR12 were used in eight 87.5 m(3) fermentation runs (four using each strain) for industrial ethanol production in an Egyptian distillery using sugar cane molasses. Mean ethanol production was 7.7% and 7.4% (w/v), respectively, with an added advantage of cooling elimination during fermentation and higher ethanol yields compared to the distillery's S. cerevisiae SIIC (ATCC 24855) strain in use. The isolate S. cerevisiae F111 was subsequently adopted by the distillery for regular production with significant economical gains and water conservation. PMID- 10797240 TI - A microscale model of bacterial and biofilm dynamics in porous media. AB - A microscale model for the transport and coupled reaction of microbes and chemicals in an idealized two-dimensional porous media has been developed. This model includes the flow, transport, and bioreaction of nutrients, electron acceptors, and microbial cells in a saturated granular porous media. The fluid and chemicals are represented as a continuum, but the bacterial cells and solid granular particles are represented discretely. Bacterial cells can attach to the particle surfaces or be advected in the bulk fluid. The bacterial cells can also be motile and move preferentially via a run and tumble mechanism toward a chemoattractant. The bacteria consume oxygen and nutrients and alter the profiles of these chemicals. Attachment of bacterial cells to the soil matrix and growth of bacteria can change the local permeability. The coupling of mass transport and bioreaction can produce spatial gradients of nutrients and electron acceptor concentrations. We describe a numerical method for the microscale model, show results of a convergence study, and present example simulations of the model system. PMID- 10797241 TI - A hollow-fiber membrane bioreactor for the removal of trichloroethylene from the vapor phase. AB - A hollow-fiber membrane bioreactor was used to separate trichloroethylene (TCE) from a gaseous waste stream with subsequent cometabolic biodegradation by a pure culture of Methylosinus trichosporium OB3b PP358. The two-stage bioreactor system was successfully operated for 20 days. PP358 was grown in a continuous-flow chemostat and circulated through the fiber lumen of a hollow-fiber membrane module (HFMM), while TCE contaminated air (141 to 191 microg/L) was pumped through the HFMM shell. Between 54% -84% TCE transfer and 92%-96% TCE cometabolism were obtained in the HFMM reactor loop. Short shell-residence times, 1.6 to 5.0 minutes, demonstrated quick throughput of TCE contaminated air. Best fit computer modeling of the biological experiments estimated mass transfer coefficients between 2.0 x 10(-3) cm/min and 5.6 x 10(-3) cm/min. The average pseudo-first-order biodegradation rate constant for the biological experiments was 0.46 L/mg TSS/d. These results demonstrate that the hollow-fiber membrane bioreactor represents an attractive technology for the bioremediation of gaseous waste streams. PMID- 10797242 TI - Production of recombinant L-leucine dehydrogenase from Bacillus cereus in pilot scale using the runaway replication system E. coli[pIET98]. AB - A method for the production of recombinant L-leucine dehydrogenase from Bacillus cereus in pilot scale is described employing the temperature induced runaway replication vector pIET98 and the Escherichia coli host strain BL21. Fed-batch cultivation using a semi-synthetic high-cell densitiy medium was adjusted in 5-L scale to yield a constant growth rate of 0,17 h(-1) and a final cell concentration of 27 g dry weight/L by exponentially increasing the nutrient supply. Runaway replication and thus, LeuDH expression was induced during the feeding phase by increasing the cultivation temperature to 41 degrees C yielding a specific enzyme activity of 110 U/mg, which corresponds to 30% of the soluble cell protein. The cultivation was terminated when the dissolved oxygen content fell below 10% saturation. The final volume activity was 600,000 U/L cultivation. No change in growth, cell density, or expression activity was observed scaling up the cultivation volume to 200 L. Thus, 120,000,000 units L-leucine dehydrogenase were obtained from one cultivation. The purification of L-leucine dehydrogenase to homogeneity was carried out by heat denaturation, liquid-liquid extraction, gel filtration, and anion-exchange chromatography to give pure enzyme in 65% yield. The integrity of the recombinant enzyme was tested measuring the molecular weight and determining the N-terminal amino acid sequence. PMID- 10797243 TI - Continuous enzymatic esterification of glycerol with (poly)unsaturated fatty acids in a packed-bed reactor. AB - Enzymatic synthesis of mono-, di-, and triacyglycerols from (poly)unsaturated fatty acids (linoleic, oleic, and conjugated linoleic acids) has been studied as a solvent-free reaction in a packed-bed reactor containing an immobilized lipase from Mucor miehei. The extents of the esterification reactions of interest are primarily determined by the molar ratio of glycerol to fatty acid because the presence of excess glycerol as a immiscible phase is responsible for reducing the activity of the water produced by the esterification reactions. For molar ratios of fatty acid to glycerol of less than 1.5, the percentage of the fatty acid esterified decreases quasi-linearly with an increase in this molar ratio. By appropriate manipulation of the fluid-residence time, one can control the relative proportions of the various acylglycerols in the effluent stream. At the outlet of the reactor, one observes excellent spontaneous separation of the glycerol and acylglycerol/fatty acid phases. At 50 degrees C and a fluid residence time of 1 hour, as much as 90% of the fatty acid can be esterified when the molar ratio of fatty acid to glycerol is 0.33 or less. PMID- 10797244 TI - Use of anion exchange resin-packed capillary column for rapid detection of anti double-stranded DNA antibody in systemic lupus erythematosus serum. AB - An anti-double-stranded (ds) DNA antibodies in serum-detection system was developed based on flow immunoassay with packed-capillary column. Alkaline phosphatase conjugated DNA (ALP-DNA) and anti-dsDNA antibody were separated from unreacted ALP-DNA on the basis of the difference in isoelectric point. A linear anti-dsDNA antibody dose-response curve was obtained between luminescence intensity and concentration of anti-dsDNA antibody in the range 25-200 IU/mL. This simple technique permits the assay of anti-dsDNA autoimmune antibodies within 7 minutes. These results give reduced detection times compared with those previously reported (Lim et al., 1999) through the use of capillary columns. PMID- 10797245 TI - Purification of a cystic fibrosis plasmid vector for gene therapy using hydrophobic interaction chromatography. AB - The success and validity of gene therapy and DNA vaccination in in vivo experiments and human clinical trials depend on the ability to produce large amounts of plasmid DNA according to defined specifications. A new method is described for the purification of a cystic fibrosis plasmid vector (pCF1-CFTR) of clinical grade, which includes an ammonium sulfate precipitation followed by hydrophobic interaction chromatography (HIC) using a Sepharose gel derivatized with 1,4-butanediol-diglycidylether. The use of HIC took advantage of the more hydrophobic character of single-stranded nucleic acid impurities as compared with double-stranded plasmid DNA. RNA, denatured genomic and plasmid DNAs, with large stretches of single strands, and lipopolysaccharides (LPS) that are more hydrophobic than supercoiled plasmid, were retained and separated from nonbinding plasmid DNA in a 14-cm HIC column. Anion-exchange HPLC analysis proved that >70% of the loaded plasmid was recovered after HIC. RNA and denatured plasmid in the final plasmid preparation were undetectable by agarose electrophoresis. Other impurities, such as host genomic DNA and LPS, were reduced to residual values with the HIC column (<6 ng/microg pDNA and 0.048 EU/microg pDNA, respectively). The total reduction in LPS load in the combined ammonium acetate precipitation and HIC was 400,000-fold. Host proteins were not detected in the final preparation by bicinchoninic acid (BCA) assay and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Plasmid identity was confirmed by restriction analysis and biological activity by transformation experiments. The process presented constitutes an advance over existing methodologies, is scaleable, and meets quality standards because it does not require the use of additives that usually pose a challenge to validation and raise regulatory concerns. PMID- 10797246 TI - Contamination of a high-cell-density continuous bioreactor. AB - Continuous fermentations were carried out with a recombinant flocculent Saccharomyces cerevisiae strain in an airlift bioreactor. Once operating under steady state at a dilution rate of 0.45 h(-1), the bioreactor was contaminated with Escherichia coli cells. The faster growing E. coli strain was washed out of the bioreactor and the recombinant, slower growing flocculating S. cerevisiae strain remained as the only species detected in the bioreactor. Flocculation, besides allowing for the realization of high-cell-density systems with corresponding unusual high productivity, may be used as a selective property for controlling some contamination problems associated with prolonged continuous operation. PMID- 10797247 TI - Developmental analysis of habituation in the Nematode C. elegans. AB - Habituation and spontaneous recovery from habituation to tap were studied across development in C. elegans. Unlike adult worms, larval worms do not consistently swim backwards to tap, but reverse half of the time and accelerate forward half of the time. In adult worms, the tap response is produced by the integration of two competing circuits: The head touch circuit, mediated by ALM and AVM sensory neurons, produces backward movement (reversals); the tail touch circuit, mediated by PLM neurons, produces forward movement (accelerations). Because the response type changes over development, habituation of each of the subcircuits was studied separately. Habituation of the head touch circuit was studied by laser ablating PLM, and habituation of the tail circuit was studied by ablating ALM. Worms were tested at six stages of development at either 10- or 60-s interstimulus intervals. All stages of development showed normal habituation and spontaneous recovery at both interstimulus intervals. PMID- 10797248 TI - Attraction of newborn rabbits to abdominal odors of adult conspecifics differing in sex and physiological state. AB - The present study aimed at assessing the ability of newborn rabbits (1-3 days) to detect and discriminate abdominal odors emitted by adult conspecifics varying in sex and physiological state. Pups were submitted to a two-choice test exclusively based on olfaction, which permitted exposure to conspecifics either during successive or simultaneous presentations. Their orientation toward either of the two stimuli was timed. These behavioral assays revealed that (a) pups display attraction to odors of the abdomen of nonlactating, nonpregnant females and males when pitted against a control stimulus; (b) pups preferred the odor of nonlactating, nonpregnant females when simultaneously presented with the odor of a male; (c) all sexually mature females (virgin, pregnant, or lactating) were attractive to pups; (d) the odor of lactating females was preferred to the odor of either nonlactating, nonpregnant or pregnant females. Thus, from the first day after birth, newborn rabbits are able to olfactorily discriminate conspecifics from cues emitted at the abdominal level and display the strongest attraction to lactating females. PMID- 10797249 TI - Shock sensitization of startle in the developing rat. AB - Rats given a series of shocks exhibit a potentiated startle response to a loud acoustic stimulus compared to nonshocked animals. Experiment 1 showed that this shock sensitization of startle, like conditioned fear potentiation of startle to discrete cues, emerges relatively late in development (i.e., 23 days of age). Although different testing procedures were used in Experiment 2, preweanling rats still failed to exhibit the shock sensitization of startle effect. The failure to observe the shock sensitization of startle effect in preweanling rats was not due to age differences in contextual conditioning produced by the shock treatment (Experiment 3). The results of this study are discussed in terms of (a) the emergence of fear potentiation of startle during development, and (b) the relation between conditioned freezing and startle potentiation. PMID- 10797251 TI - Body scaling of grip configurations in children aged 6-12 years. AB - This study examined the influence of body scale on the grip configurations used by young children (6-12 years old) to displace cubes that varied systematically in size (L) and mass (M). It was determined if the scaling relation we had developed for adults K = log Lc + ((log Mc)/(a + bLh + cMh)) where L(c) and M(c) is the length side and mass of the cubes, respectively, and L(h) and M(h) are the length and mass of the hand, respectively, it would also predict the change in grip configurations across this age range in a fashion consistent with the principle of similitude. The statistical and scaling analyses revealed that the invariant body-scaled relation that specified the adult grip transitions also held for the 6- to 12-year-old children. PMID- 10797250 TI - A longitudinal study of hand preference in chimpanzees (Pan troglodytes). AB - A longitudinal study of hand preferences was assessed in a sample of 53 captive chimpanzees. Four measures of laterality assessed during the first 3 months of life were correlated with three measures of hand preferences assessed when the subjects were between 2 and 5 years of age. In addition, the effect of rearing environment on juvenile hand preferences was assessed in a larger sample of 83 chimpanzees. Overall, some early asymmetries were predictive of juvenile hand preferences, notably head orientation and hand-to-hand activities, and a defensive grasping response. No significant effects of rearing on hand preferences were found but males were more right-handed than females for two of the three juvenile measures. The results are discussed within the context of different developmental models of hand preference in humans. PMID- 10797252 TI - Acquisition of upper body stability during walking in toddlers. AB - This study examines the development of head and trunk movements in toddlers as they begin to walk independently. The data are from a longitudinal study of 7 infants observed from the onset of walking over a period of 46-80 weeks. Head and trunk rotations were measured in the frontal and sagittal planes together with global gait parameters (progression velocity, step cadence, length and width, duration of double support phase). The results showed that during the first weeks of walking head and trunk oscillations significantly decreased, indicating that considerable progress is made in upper body stabilization. Dramatic changes in global gait parameters also occurred at this time. After this first period of rapid changes, gait parameters continued the same developmental trend but with slower changes. The close relation between gain in head and trunk stability and improvement in walking efficacy is discussed on the basis of the individual developmental trends. PMID- 10797253 TI - EEG correlates of the development of infant joint attention skills. AB - The development of the capacity for social attention coordination, or "joint attention," is a major milestone of infancy. Data from a recent study of handicapped infants have raised the hypothesis that the tendency to initiate bids for joint attention may reflect processes associated with the frontal cortex to a greater extent than other forms of infant attention coordination (R. Caplan et al., 1993). This hypothesis was examined in a longitudinal study of 32 normally developing infants. The results indicated that EEG data at 14 months indicative of left frontal, as well as left and right central cortical activity, was associated with the tendency to initiate joint attention bids (IJA) at 14 and 18 months. In contrast, a pattern of left parietal activation and right parietal deactivation at 14 months was associated with the development of the capacity to respond to the joint attention bids (RJA) of others at 14 and 18 months. These results were interpreted to be consistent with a general anterior-posterior model of attention development (M. Posner & S. Petersen, 1990). The implications of these results for current conceptualizations of joint attention development, as well as for understanding the disturbance of joint attention skill development in autism are discussed. PMID- 10797254 TI - BRCA2 protein expression in sporadic breast carcinoma with or without allelic loss of BRCA2. AB - To elucidate the cellular role of BRCA2 in sporadic breast tumors, we studied the cellular localization and the expression of BRCA2 in carcinomas presenting or not allelic loss of BRCA2. The breast tumors were first classified with or without allelic loss of BRCA2 and then immunohistochemical staining was performed on tumors and matched normal tissues using antibodies raised against BRCA2. We showed that BRCA2 is found either in the nucleus or in perinuclear compartments such as the endoplasmic reticulum and the Golgi vesicles. We have earlier demonstrated the presence of BRCA1 as an exocrine secretion in the lumen of ductules in the normal mammary gland, as well as BRCA1 and BRCA2 in milk-fat globules inside mammary-gland ductules during lactation. Here we show that BRCA2 is present within the lumina of breast ductules, indicating that BRCA2 protein may also be secreted in the mammary gland. No correlation was found in breast tumors between the expression of BRCA2 protein and allelic loss of BRCA2. PMID- 10797255 TI - Endocrine host responses during early and late phases of tumor development. AB - It is well established that hormones affect tumor growth. Conversely, inoculation of cells obtained from tumors that had been transplanted for many generations causes changes in the concentration of different hormones before and after tumor detection. We aimed at answering the question of whether hormonal alterations also occur during the development of primary tumors and following transplantation of tumors from early generations. Primary tumors were induced in mice by either the carcinogenic agent 3-methylcholanthrene, which produces fibrosarcomas, or the milk-transmitted mammary tumor virus, which induces adenocarcinomas. The results showed that (i) in both models, an early reduction in plasma insulin and prolactin levels occurred, and in the case of insulin, this reduction was sustained for a prolong period prior to tumor detection, indicating that recognition by the host of emergent tumor cells triggers an endocrine response; (ii) in contrast with multiply transplanted tumors, cells from early transplant generations produced no significant endocrine changes during latency; (iii) irrespective of whether they were primary or transplanted, large tumor burdens caused similar hormonal alterations, consisting of increased corticosterone and growth hormone and decreased insulin, thyroxin, prolactin and sex steroid levels in blood. Our comprehensive longitudinal study demonstrates host endocrine responses during different stages of neoplastic development. PMID- 10797256 TI - Apoptosis-inducing levels of UV radiation and proteasome inhibitors produce opposite effects on p21(WAF1) in human melanoma cells. AB - The stability of p21(WAF1) and p53 is increased by UV radiation or proteasome inhibitors in normal and some tumor cells. However, p21(WAF1) can either stimulate in vitro assembly of active cyclin-kinase complexes at low concentrations or inhibit this activity at high concentrations. Also, ectopic p21(WAF1) over-expression has been reported to promote or suppress apoptosis, depending on the target cells. We have investigated changes in p21(WAF1) expression as a result of exposure to either 25 J/m(2) UV or 10 microM MG-115 proteasome inhibitor, both of which cause apoptosis in human C8161 melanoma cells. p21(WAF1) mRNA increased in response to UV irradiation but failed to accumulate at the protein level because of its early UV-activated degradation counteracted by proteasome inhibition. UV-mediated loss of p21(WAF1) protein preceding induction of p53 and cell death was greater in non-metastatic than in metastatic C8161 melanoma cells. No loss in p21(WAF1) occurred with apoptosis induced by 10 microM proteasome inhibitors MG-115 or lactacystin, mediated by over-expression of p21(WAF1). Our results suggest that conditions causing prolonged or permanent changes in basal levels of p21(WAF1) may impair its reversible cell-cycle checkpoint function, leading to irreversible growth arrest or cell death. PMID- 10797257 TI - Tamoxifen-resistant glioma-cell sub-populations are characterized by increased migration and proliferation. AB - Multifocal tumor recurrence of glioblastomas occurs in up to 14% of patients. In a parallel phase-II-study investigating post-operative treatment with tamoxifen (TAM), carboplatin and radiation therapy for glioblastomas, 16 of 49 patients (33%) showed multifocal recurrence, which developed after a mean of 46 weeks, raising the question of an association with therapy. We studied the interrelation of proliferation and migration in the presence of different protein-kinase-C(PKC) inhibitors (TAM, staurosporine, hypericin) in 2 glioma cell lines. In addition, 3 cell lines were selected for TAM resistance by repeated cycles of treatment with sub-lethal concentrations of TAM. The proliferative capacity and the invasive potential of selected sub-populations were assessed using growth-curve experiments, monolayer migration, and cell-adhesion assays. Treatment with all PKC inhibitors tested resulted in a dose-dependent decrease of proliferation, while motility was altered only at significantly higher doses. Resistance to TAM occurred in all 3 selected cell lines. The TAM-resistant sub-populations showed significantly increased proliferation, migration and adhesion as compared with the parental (non-selected) cell line. The higher incidence of multifocal disease after TAM treatment was paralleled by increased migratory potential of TAM treated cells in vitro. PMID- 10797258 TI - Identification of a unique binding protein specific for a novel retinoid inducing cellular apoptosis. AB - The retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylic acid (AHPN, CD437) induces apoptosis in a variety of cell types, many of which are cancer cells that resist the antiproliferative and/or differentiating effects of retinoids. While the retinoids exert their effects by binding to the retinoic acid nuclear receptors (RARs) or retinoid X receptors (RXRs), AHPN (CD437) binds to another protein with different ligand specificity. In nuclear extracts from HL 60R cells the binding of AHPN (CD437) was only minimally competed by either retinoic acid (tRA)or 9-cis-retinoic acid (9-cis-RA), the natural ligands for the RARs and RXRs, respectively. Moreover, AHPN (CD437) was unable to compete with either tRA or 9-cis-RA for binding to endogenous retinoid receptors in nuclear extracts from the MDA-MB-468 breast carcinoma cell line. Size exclusion chromatography revealed AHPN binding to a 95 kDa protein(s) which is neither an RAR or RXR. Our results suggest that apoptosis induction by AHPN (CD437) may occur through interaction with another protein and is independent of the RAR/RXR signaling pathways. PMID- 10797259 TI - NE-dlg, a mammalian homolog of Drosophila dlg tumor suppressor, induces growth suppression and impairment of cell adhesion: possible involvement of down regulation of beta-catenin by NE-dlg expression. AB - Membrane-associated guanylate kinases (MAGUKs) are known to function as scaffolds for forming multiprotein complexes at the synaptic junctions of neuronal cells and at sites of epithelial cell-cell contact. In Drosophila, mutations of the lethal (1)-discs large (dlg) gene, which encodes a MAGUK protein, leads to post synaptic structure defects in neuronal cells and neoplastic overgrowth of epithelial cells. We previously showed that NE-dlg (neuronal and endocrine dlg), a human homolog of the dlg, plays a crucial role in formation of synaptic structure in human neuronal cells. Here we demonstrate that NE-dlg, similar to Drosophila dlg, is involved in regulation of cell cycle progression and adhesive ability of non-neuronal cells. Overexpression of NE-dlg in proliferating cells including various cancer cell lines induced growth suppression and impairment of cell adhesive ability. Furthermore, NE-dlg overexpression caused the down regulation of beta-catenin in cancer cells regardless of mutations in the APC (adenomatous polyposis coli) gene. The PDZ domains of NE-dlg were found to be essential for the growth suppression, loss of adhesive property and down regulation of beta-catenin. We propose that NE-dlg regulates the cell growth and adhesive ability by controlling the level of beta-catenin through an APC independent pathway. Inactivation of NE-dlg may therefore contribute to development and/or progression of human neoplasms. PMID- 10797260 TI - Expression of stress protein gp96, a tumor rejection antigen, in human colorectal cancer. AB - Preparations of stress protein gp96 from tumor cells are active as tumor vaccines by eliciting immune responses against mixtures of individual tumor peptide antigens which are complexed to gp96. Due to the individual antigenicity of tumors, a vaccine consisting of tumor-derived gp96 has to be prepared individually for each patient from autologous tumor tissue. So far, gp96 expression by human tumors has not been analyzed. Here, we report stable and mostly homogenous expression of gp96 by colorectal cancer, which was enhanced compared to surrounding tumor stroma in 70% to 80% of colorectal cancer specimens. Fewer non-metastatic than metastatic primary cancer specimens showed enhanced gp96 expression. Glucose deprivation increased gp96 protein and RNA expression in the human colon cancer cell line HT-29 in accordance with the role of gp96 as a glucose-regulated stress protein. Additionally, TNF-alpha, interferons and other cytokines induced an increase of gp96 RNA expression in HT 29 cells, suggesting that gp96 expression by colorectal cancer cells can be influenced by different methods of immunomodulation. The stable and homogenous expression of gp96 in 19 primary and metastatic colorectal cancer specimens and the up-regulation of gp96 in colon cancer cells by glucose deprivation point to an essential role of this stress protein in colorectal cancer, presumably by protecting against hostile conditions of the tumor micro-environment like glucose deprivation. In view of these results, loss of gp96 expression by colorectal cancer cells as an immune escape mechanism is unlikely. PMID- 10797261 TI - Genetic changes in breast cancer detected by comparative genomic hybridisation. AB - Breast cancer is characterised by a number of genetic aberrations. Our purpose was to use comparative genomic hybridisation (CGH) to screen breast carcinomas for copy number changes: 44 ductal and 8 lobular carcinomas were studied and a large number of genetic aberrations identified. Many of these showed similarity to previous CGH results, however, a number of loci not previously shown to have undergone frequent change were identified. This included copy number gains affecting chromosomes 1p, 4q, 5q, 6q and 13q. Furthermore, we have identified 2 regions of copy number change, the gain on 5p and deletion of 16q, which correlated with lobular carcinomas. Our results highlight several areas of the genome that may be important in the molecular genetics of breast cancer. PMID- 10797262 TI - Loss of heterozygosity at chromosome segments 8p22 and 8p11.2-21.1 in transitional-cell carcinoma of the urinary bladder. AB - To identify the putative tumor-suppressor gene (TSG) involved in transitional cell carcinoma (TCC) of the urinary bladder, we undertook an allelotyping analysis in 48 cases of TCC. Relatively high percentages of allelic loss were found in 2p (5 of 23, 21.7%), 8p (9 of 21, 42.9%), 9p (4 of 20, 20.0%), 12q (6 of 28, 21.4%), 15q (1 of 5, 20%; 4 of 20, 20%), 17p (7 of 26, 26.9%) and 22q (6 of 23, 26.1%). On the basis of these results, fine-deletion mapping was performed on chromosome 8 in 52 cases by PCR of 15 microsatellite markers. Two distinct regions of common deletion were found. A 10 cM telomeric region was located to 8p22, defined by D8S511 and D8S258. A 17 cM centromeric region was located to 8p11.2-21.1, flanked by D8S298 and D8S535. The distance between the telomeric and the centromeric regions of common deletion was 3 cM. Loss of heterozygosity of 8p22 was frequently observed in tumors of high grade or advanced stage. PMID- 10797263 TI - Alu-associated interstitial deletions and chromosomal re-arrangement in 2 human multidrug-resistant cell lines. AB - Previous studies have shown that gene re-arrangements play a significant role in tumorigenesis. Gene re-arrangements involving the human multidrug resistance-1 (MDR1) gene have been identified as a mechanism for MDR1 over-expression in human malignant cells. In 2 multidrug-resistant human cancer sublines with high levels of MDR1 and P-glycoprotein (MCF7/TX400 and S48-3s/Adr10), hybrid mRNAs containing sequences from MDR1 and an unrelated gene have previously been identified. To characterize and determine the site of the re-arrangements resulting in generation of hybrid mRNAs, we first constructed a lambda phage library extending over a contiguous genomic region of 100 kb and containing the region upstream of MDR1. In MCF7/TX400 cells, homologous recombination was observed involving an Alu repeat 80 kb upstream of the MDR1 gene, with a 79 bp intra-Alu deletion flanked by chi-like sequences at the re-arrangement junction. By contrast, non-homologous recombination was observed in S48-3s/Adr10 cells with Alu repeats near the junction sequence. While the specific features of the breakpoints appear to be different, Alu repeats might be involved in both gene re-arrangements. The gene re-arrangements at or near the Alu sequence should be regarded as potentially involved in the transcriptional activation of human MDR1. PMID- 10797264 TI - Gene expression profiling of advanced lung cancer. AB - Lung cancer is a highly aggressive neoplasm with 85% mortality. To identify new tumor-associated genes, we compared the expression profile of a primary metastasizing adenocarcinoma with normal airway epithelial cells. Two cDNA libraries of up- and downregulated genes were generated, comprising 253 and 299 clones, respectively. The sequence analysis revealed 205 different known genes and 314 cDNA fragments of unknown functions. Northern-blot analysis of 167 clones confirmed differential expression in 58%, and indicated a similar expression pattern in additional lung-cancer cell lines for selected clones, strengthening the value of this model for the identification of new candidate genes in lung carcinogenesis. PMID- 10797265 TI - Comparison of chromosome 3p deletions between cervical precancers synchronous with and without invasive cancer. AB - Cervical cancers are considered to originate from a series of pre-malignant lesions (cervical intra-epithelial neoplasia, CIN). The mechanisms behind these events are unknown. In addition to HPV infection, deletions of chromosome 3p have been found to be a frequent event in cervical cancer and likely play an important role in the transition of CIN to invasive cancer. To classify the potential role of 3p deletions in early-stage cervical carcinogenesis, we analyzed LOH of 3p in cervical precancers. Thirty cases with single or multiple CIN lesions were selected for the study, including 20 cases without and 10 cases with synchronous invasive cancers. Allelic losses on 1 or more 3p loci were recorded in 33% (3/9) of CIN II and 36% (5/14) of CIN III lesions from 20 cases without co-existing invasive cancer, whereas an increasing percentage of LOH was observed in the 10 precancerous lesions synchronous with invasive cancer, with 71% (5/7) CIN II and 76% (13/17) CIN III lesions. This result implies that 3p deletions have selective roles in early transition of pre-malignancy to invasive cancer. Comparing the LOH patterns between the 2 groups, genetic deletions in cases with invasive cancers involved extensive regions of 3p but were more localized in precancer cases without concomitant invasive cancer. Two interstitial regions, 3p22-21.3 around marker D3S1260 and 3p21.1 around markers D3S1289 and D3S1076, were most frequently deleted in both groups, suggesting that these 2 regions are novel tumor-suppressor loci which may play a role in early transition of cervical precancer to invasive cancer. Identical LOH patterns between multiple CIN lesions and synchronous invasive cancer in the same case suggests that different cervical precancers and invasive cancer are genetically linked and most likely originate from a single precursor cell. PMID- 10797266 TI - A human melanoma metastasis-suppressor locus maps to 6q16.3-q23. AB - Loss, deletion or rearrangement along large portions of the long arm (q-arm) of chromosome 6 occurs in >80% of late-stage human melanomas, suggesting that genes controlling malignant characteristics are encoded there. Metastasis, but not tumorigenicity, was completely suppressed in the human melanoma cell line C8161 into which an additional intact chromosome 6 had been introduced by microcell mediated chromosome transfer. Our objective was to refine the location of a putative metastasis suppressor gene. To do this, we transferred an intact (neo6) and a deletion variant [neo6qdel; neo6(del)(q16.3-q23)] of neomycin-tagged human chromosome 6 into metastatic C8161 subclone 9 (C8161.9) by MMCT. Single cell hybrid clones were selected in G-418 and isolated. Following verification that the hybrids retained the expected regions of chromosome 6 using a panel of polymorphic sequence-tagged sites, the hybrids were tested for tumorigenicity and metastasis in athymic mice. As reported previously, intact, normal chromosome 6 suppressed metastasis whether tumor cells were injected i.v. or into an orthotopic (i.e., intradermal) site. In contrast, metastasis was not suppressed in the neo6qdel hybrids. Tumorigenicity was unaffected in hybrids prepared with either chromosome 6 donor. These data strongly suggest that a human melanoma metastasis suppressor locus maps between 6q16.3-q23 ( approximately 40 cM). PMID- 10797267 TI - Germline mutations of the dpc4 gene in Korean juvenile polyposis patients. AB - Juvenile polyposis is an uncommon condition characterized by the development of multiple (usually more than 5) juvenile polyps in the gastrointestinal tract, especially in the colon. This disease usually occurs during childhood, and is inherited in an autosomal dominant fashion. It has been suggested that the dpc4 (deleted in pancreatic carcinoma, locus 4) gene, which is located on chromosome 18q21.1, might cause juvenile polyposis. The dpc4 (smad4) gene is a candidate tumor-suppressor gene and may play a role in the TGF-beta-signaling pathway. To confirm the idea that alterations of the dpc4 gene may result in juvenile polyposis, we screened 5 Korean juvenile-polyposis patients by PCR-SSCP (single strand conformation polymorphism) analysis and bi-directional sequencing. There were germline mutations of the dpc4 gene in 3 out of the 5 patients: 2 had a genetic alteration in exon 9 and the third had a mutation in exon 8. These germline mutations occurred in the C-terminus of the dpc4 gene, similar to most published mutations. One patient exhibited a non-sense mutation (codon 388), which changed a glutamine codon (CAG) to a stop codon (TAG). The second patient harbored a mis-sense mutation (codon 390), causing a non-conservative amino-acid change . The third patient had a mis-sense mutation in exon 8 (codon 361), which altered an arginine codon (CGC) into a histidine codon (CAC). PMID- 10797268 TI - Genetic alterations involving exon 3 of the beta-catenin gene do not play a role in adenocarcinomas of the esophagus. AB - Beta-catenin has been identified as an oncogene. Phosphorylation of sites encoded by exon 3 of the beta-catenin gene facilitates degradation of this protein by the adenomatous polyposis coli (apc) gene product. Mutations in these sites or inactivation of apc lead to stabilization of beta-catenin, which then translocates to the nucleus where it modulates the transcription of genes involved in tumor formation. To explore the role of beta-catenin mutations in adenocarcinomas of the esophagus, we screened for genetic alterations in exon 3 in 69 tumor samples. We detected no mutations in exon 3 by PCR-SSCP analysis nor did we find large interstitial deletions involving exon 3. beta-catenin immunostaining on 54 tumors showed focal nuclear staining in 7 tumors and homogeneous nuclear staining in 3 tumors; in the latter; no mutations in the mutation cluster region of apc were detected. These results show that genetic alterations of exon 3 of the beta-catenin gene do not occur and therefore do not contribute to the pathogenesis of esophageal adenocarcinomas. The abnormal cytoplasmic and nuclear localization of beta-catenin indicates that other mechanisms leading to elevated free beta-catenin in these cancers must be involved. PMID- 10797269 TI - Clonal expansion of Melan A-specific cytotoxic T lymphocytes in a melanoma patient responding to continued immunization with melanoma-associated peptides. AB - Peptides derived from human tumor antigens have been used in a number of clinical trials to induce specific immune responses against autologous tumors in cancer patients. Although favorable clinical results were observed in single patients, immune responses correlating with tumor regression were either not detected or in case of responses, the T-cell specificity was difficult to demonstrate. In this study, we analyzed antigen-specific T-cell responses induced in the skin and in peripheral blood lymphocytes (PBL) in an HLA-A2-positive melanoma patient. The patient showed major regression of metastatic melanoma under continued immunization with peptides derived from the melanocyte differentiation antigens Melan A/MART-1, tyrosinase and gp100/Pmel17. Based on the identification of different T-cell receptor (TCR) families reactive with Melan A/MART-1, we have demonstrated that i.d. immunization with peptides alone leads to oligoclonal expansion of Melan A/MART-1-specific cytotoxic T lymphocytes (CTL), detectable in local delayed-type hypersensitivity (DTH) reactions and PBL. A monoclonal expansion of a Melan A/MART-1-specific TCR VB 16 CTL was reproducibly observed after in vitro stimulation with Melan A/MART-1 peptides. The same TCR VB 16 CTL clone was detected in skin biopsies taken from vitiligo areas. Our findings provide strong evidence for the effective induction of specific T-cell responses to Melan A/MART-1 by i.d. immunization with peptide alone, which accounts for dermal depigmentation, specific cytotoxicity against Melan A/MART-1-expressing melanoma cells and clinical tumor regression. PMID- 10797270 TI - Frequent lack of translation of antigen presentation-associated molecules MHC class I, CD1a and Beta(2)-microglobulin in Reed-Sternberg cells. AB - Epstein-Barr virus (EBV) is present in Reed-Sternberg (RS) cells of a substantial proportion of Hodgkin's lymphoma cases. Most EBV-positive cases are also MHC class I-positive, whereas the majority of EBV-negative cases lack detectable levels of MHC class I expression. Application of the SAGE technique has led to the identification of tags corresponding to MHC class I and beta(2)-microglobulin genes in the EBV- and MHC class I-negative L428 Hodgkin's cell line. Further expression studies indicated that single RS cells that do not express HLA class I also lack beta(2)-microglobulins but frequently contain mRNA coding for these proteins. Another tag was identified corresponding to CD1a, a thymocyte and Langerhans cell antigen structurally related to the MHC class I genes. CD1a expression studies revealed mRNA in all cell lines and in several of the single cells, whereas immunostaining showed a cytoplasmic signal in only 2 of the 4 cell lines and in none of the Hodgkin's lymphoma tissue samples. In conclusion, RS cells frequently lack MHC class I, beta(2)-microglobulin and CD1a protein expression but contain mRNA coding for these proteins in some of the RS cells, suggesting a common mechanism affecting the translation of these antigen presentation-associated molecules. PMID- 10797271 TI - Apoptosis induction and cyclooxygenase-2 regulation in human colorectal adenoma and carcinoma cell lines by the cyclooxygenase-2-selective non-steroidal anti inflammatory drug NS-398. AB - We determined the effect of the highly selective cyclooxygenase-2 (COX-2) inhibitor NS-398 on proliferation, apoptosis and COX-2 regulation in 3 pre malignant human colorectal adenoma cell lines (RG/C2, AA/C1, RR/C1) and compared its effect on 3 colorectal carcinoma cell lines (HT29, KS, JW2). COX-2 protein was expressed in each cell line derived from an adenoma, thus providing evidence that COX-2 is expressed in the tumour cells themselves at an early stage in human colorectal adenoma formation. NS-398 (20 to 100 microM for 96 h) induced apoptosis and inhibited the proliferation of the adenoma cell lines. Of the 3 carcinoma lines, only HT29 expressed COX-2 protein, yet each line was similarly sensitive to NS-398. There was a positive correlation between overall sensitivity of the cell lines (determined by the attached cell yield) and sensitivity to NS 398-induced apoptosis, suggesting that apoptosis is the dominant anti proliferative effect of NS-398. Two of the 3 adenoma cell lines (RG/C2, AA/C1) were less sensitive than the carcinoma cell lines. NS-398 up-regulated COX-2 protein expression in the HT29 and adenoma cell lines. This was studied further in HT29 cultures, where treatment with NS-398 inhibited COX-2 activity, reducing prostaglandin E(2) secretion. Here, neither the increase in COX-2 protein expression nor the anti-proliferative and apoptosis-inducing effect of NS-398 was prevented by addition of exogenous prostaglandin E(2). Apoptosis appears to be the dominant anti-proliferative effect of NS-398 and, in COX-2 expressing cells, may be mechanistically linked to the observed induction of COX-2 protein expression upon treatment with NS-398. PMID- 10797272 TI - A recombinant cytotoxic chimera based on mammalian deoxyribonuclease-I. AB - A number of mammalian proteins with suitable biological activities have been considered for use in targeted tumour therapy. Deoxyribonuclease-I (DNase-I), an endonuclease that degrades double-stranded DNA, represents an attractive candidate for tumour targeting since it is normally non-toxic yet could be highly cytotoxic when redirected to the cell nucleus. Our aim was to investigate the cytotoxic potential of mammalian DNase-I and its possible use in tumour-targeting strategies for cancer therapy. A chimeric molecule comprising a scFv reactive against the human placental alkaline phosphatase (hPLAP) and bovine pancreatic DNase-I was designed and investigated. The development of a tightly controlled system for the bacterial expression of DNase-I and its chimera is described. The production and purification of active DNase-I from the soluble cell fraction and significant yields from the insoluble fraction by isolation and refolding are described. The construction, expression, purification and in vitro characterisation of an anti-PLAP scFv-DNase-I chimera is also described. This molecule was shown to possess both antigen-binding and DNA-degrading activity in in vitro assays, thus combining the specific cell-targeting properties of the scFv and the potent, highly catalytic activity of the endonuclease. Furthermore, this chimeric molecule was highly cytotoxic in vitro in cells expressing the PLAP antigen. Targeting mammalian DNase-I provides a novel therapeutic strategy for selective cell killing, with the promise of less systemic toxicity and immunogenicity than currently used immunotoxins. PMID- 10797273 TI - Induction of apoptosis in lung-cancer cells following bcl-xL anti-sense treatment. AB - Over-expression of the anti-apoptotic protein bcl-xL is frequently found in lung cancer where it potentially contributes to tumor development, progression and drug resistance. To override the apoptotic block in lung-adenocarcinoma and small cell-lung-cancer (SCLC) cells caused by over-expression of bcl-xL, an anti-sense oligodeoxynucleotide was designed targeting a sequence unique to the bcl-xL coding region and not shared by the pro-apoptotic splice variant bcl-xS. Moreover, to improve the biophysical properties of the anti-sense compound, 2; methoxy-ethoxy modifications were made to selected deoxy-ribose residues. The resulting gapmer oligonucleotide 4259 was tested on lung-adenocarcinoma and SCLC cell lines in vitro. Treatment of the adenocarcinoma cell lines A549 and NCI-H125 and the SCLC cell lines SW2 and NCI-H69 with 600 nM 4259 reduced bcl-xL levels by 70 to 90%. In the lung-adenocarcinoma cell lines, apoptosis was induced, as indicated by caspase-3-like protease activation and nuclear condensation and fragmentation. In contrast, in the SCLC cell lines, no induction of apoptosis could be demonstrated. These findings imply that bcl-xL is a more critical survival factor for lung adenocarcinomas than for SCLC, and suggest the use of oligonucleotide 4259 for therapy of this major sub-type of lung cancer. PMID- 10797274 TI - Down-regulation of trypsinogen-2 expression by chemically modified tetracyclines: association with reduced cancer cell migration. AB - Many types of human tumor express trypsinogen-2, which may be a significant factor in the activation of pro-MMPs and the invasiveness of tumors. Prevention of trypsinogen-2 expression in cancer cells might be of benefit in cancer therapy. We describe here chemicals capable of down-regulating the expression of trypsinogen-2. Doxycycline (DOXY) and chemically modified tetracyclines (CMTs), previously known as inhibitors of the matrix metalloproteinase (MMP)-dependent proteinase cascade, down-regulated the mRNA and protein expression of trypsinogen 2 by COLO-205 human colon adenocarcinoma cells at therapeutically attainable concentrations (0. 1 to 1.0 microM). DOXY specifically inhibited the activation of pro-MMP-9 and cell migration induced by enteropeptidase, a specific activator of trypsinogen. Pro-MMP-9 activation and cell migration were also inhibited by tumor-associated trypsin inhibitor (TATI), which is a highly specific inhibitor of trypsin. CMT-3 as well as CMT-5 also inhibited cell migration, but an effect on the enteropeptidase-enhanced activation of pro-MMP-9 was not observed. Our results indicate that CMTs, DOXY and TATI inhibit cancer cell migration by down regulating trypsinogen-2 expression or activity. Inhibition of trypsinogen-2 expression may represent a mechanism contributing to the ability of CMTs to suppress the pericellular proteolytic activity of some tumors. PMID- 10797275 TI - Genetic grafting of membrane-acting peptides to the cytotoxin dianthin augments its ability to de-stabilize lipid bilayers and enhances its cytotoxic potential as the component of transferrin-toxin conjugates. AB - Three chimeric proteins were obtained by fusing together the dianthin gene and DNA fragments encoding for the following membrane-acting peptides: the N-terminus of protein G of the vesicular stomatitis virus (KFT25), the N terminus of the HA2 hemagglutinin of influenza virus (pHA2), and a membrane-acting peptide (pJVE). Chimeric dianthins (KFT25DIA, pHA2DIA and pJVEDIA) retained full enzymatic activity in cell-free assays and showed increased ability to induce pH-dependent calcein release from large unilamellar vesicles (LUVs). pHA2DIA and pJVEDIA also showed faster kinetics of interaction with LUVs, while KFT25DIA and pHA2DIA displayed a reduced cytotoxicity as compared to wild-type dianthin. Conjugates made by chemically cross-linking KFT25DIA or pJVEDIA and human transferrin (Tfn) showed greater cell-killing efficiency than conjugates of Tfn and wild-type dianthin. As a consequence, by fusion of membrane-acting peptides to the dianthin sequence the specificity factor (i.e., the ratio between non-specific and specific toxicity) of Tfn-KFT25DIA, Tfn-pHA2DIA and Tfn-pJVEDIA was increased with respect to that of Tfn-based conjugates made with wild-type dianthin. Taken together, our results suggest that genetic fusion of membrane-acting peptides to enzymatic cytotoxins results in the acquisition of new physico-chemical properties exploitable for designing new recombinant cytotoxins and to tackle cell-intoxication mechanisms. PMID- 10797276 TI - Survival in operable non-small-cell lung cancer: role of p53 mutations, tobacco smoking and asbestos exposure. AB - Validated markers are needed to identify operable lung cancer patients with poor prognosis. About one-half of non-small-cell lung cancers (NSCLCs) carry a mutation in the p53 tumor-suppressor gene. We examined 101 NSCLC patients for surgical stage, completeness of resection, tobacco smoking, asbestos exposure, age, gender and p53 gene mutations as prognostic factors after a follow-up period of 4 years. Cox's multivariate regression model was applied to quantify the associations with overall and cancer-related survival. Patients with a wild-type p53 gene had an overall 4-year survival of 43% and those with a mutated p53 gene, 35%. In squamous-cell carcinoma, stage and heavy smoking, defined as the median of pack-years smoked, had prognostic significance for overall survival. Only stage was associated with poor cancer-related survival. Asbestos exposure was not associated with overall survival or cancer-related survival in squamous-cell carcinoma or adenocarcinoma. In adenocarcinoma, p53 mutation, in addition to stage, emerged as a significant predictor of poor cancer-related survival. PMID- 10797277 TI - Does gender of offspring modify the time-related effects of a pregnancy on breast cancer risk? PMID- 10797278 TI - Nitrate, nitrite and N-nitroso compounds in human bladder cancer associated with schistosomiasis. PMID- 10797279 TI - Dietary exposure to nitrite and nitrosamines and risk of nasopharyngeal carcinoma in Taiwan. AB - Previous studies of nasopharyngeal carcinoma (NPC) have found elevated risks with higher consumption of salted fish and preserved foods, particularly during childhood. These foods can contain high levels of nitrosamines; however, most studies have not estimated exposure to nitrosamines directly. We conducted a case control study in Taiwan to evaluate dietary intakes and NPC risk. A total of 375 cases (99% response rate) and 327 controls (88% response rate) were interviewed about their diet as an adult and at age 10 using a food-frequency questionnaire. We interviewed mothers of participants about their child's diet at age 10, age 3 and during weaning and the mother's diet while she was breast-feeding. Mothers of 96 cases and 120 controls were interviewed. Nitrosamine and nitrite levels were assigned to 66 foods based on published values. Intake of nitrosamines and nitrite as an adult was not associated with risk of NPC. High intakes of nitrosamines and nitrite during childhood and weaning were associated with increased risks of NPC for foods other than soy products. Adjusted odds ratios for the highest quartile were 2.2 [95% confidence interval (CI) 0.8-5.6] for age 10, 2.6 (95% CI 1.0-7.0) for age 3 and 3.9 (95% CI 1.4-10.4) for weaning diet. Intakes of nitrite and nitrosamines from soybean products during childhood and weaning were inversely associated with risk. Soybeans contain known inhibitors of nitrosation, and thus may explain the inverse association we observed. Our results suggest that nitrosamine and nitrite intake during childhood may play a role in the development of NPC. PMID- 10797280 TI - Expression of CYP1A1, CYP1B1 and CYP3A, and polycyclic aromatic hydrocarbon-DNA adduct formation in bronchoalveolar macrophages of smokers and non-smokers. AB - Variability in the expression of enzymes metabolizing carcinogens derived from cigarette smoke may contribute to individual susceptibility to pulmonary carcinogenesis. This study was designed to determine the effects of smoking and 3 major cytochrome P450 (CYP) enzymes, i.e., CYP1A1, CYP1B1 and CYP3A, which metabolize polycyclic aromatic hydrocarbons (PAH) on PAH-DNA adduct formation in the bronchoalveolar macrophages (BAM) of 31 smokers and 16 non-smokers. CYP protein levels were determined by immunoblotting and PAH-DNA adduct levels by the nuclease P1 enhanced (32)P-postlabeling method. The expression of specific CYP forms was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) from 10 additional samples. CYP3A protein, CYP3A5 by RT-PCR, was detected in the majority of samples from smokers and non-smokers. The levels of CYP3A appeared to be lower in active smokers than in ex-smokers (p = 0.10) or never smokers (p = 0.02). CYP1A1 was not detectable by either immunoblotting or RT-PCR. The expression of CYP1B1 was low or undetectable in most samples. The PAH-DNA adduct levels were higher (mean 1.57/10(8) nucleotides) in samples from smokers compared with non-smokers (mean 0.42/10(8) nucleotides, p < 0.001) and the number of adducts correlated with the number of cigarettes smoked daily (regression analysis, p < 0. 001). Higher levels of adducts were detected in samples from smokers with a high level of CYP3A compared with those with a low level (regression analysis, p = 0.002). As CYP3A5 is abundant in both lung epithelial cells and BAM, its association with adduct formation suggests that this CYP form may be important in the activation of cigarette smoke procarcinogens. PMID- 10797281 TI - Cell death induced by TNF or serum starvation is independent of ErbB receptor signaling in MCF-7 breast carcinoma cells. AB - The ErbB receptor tyrosine kinase family consists of the epidermal growth factor (EGF) receptor (ErbB1) and three related receptors (ErbB2, ErbB3, ErbB4). Their intrinsic tyrosine kinases can be activated by receptor-dimerization induced by numerous ligands or overexpression. ErbB receptors are frequently overexpressed in breast cancer, and their overexpression is associated with protection from apoptosis. To directly assess their role in apoptosis sensitivity of breast cancer cells, we established MCF-7 breast carcinoma cell lines overexpressing each ErbB receptor alone or in all possible pairs. Overexpression of ErbB1, ErbB2 and ErbB4 receptors was enough to activate them as judged by their phosphorylation, whereas co-expression of other ErbB receptors was necessary for the phosphorylation of the ErbB3. Surprisingly, overexpression of the ErbB receptors even when combined with treatment with their ligands (EGF, transforming growth factor alpha, betacellulin, neuregulins) failed to protect the MCF-7 cells from cell death induced by either tumor necrosis factor (TNF) or serum starvation. During starvation TGF-alpha, however, increased the cell size of the ErbB1 overexpressing cell line, and neuregulin1-beta1 increased that of all cell lines. In conclusion, our data does not support the role of ErbB receptors in the regulation of cell death induced by TNF or serum starvation, and the observed association in breast cancer may be due to other concomitant changes. PMID- 10797282 TI - Role of macronutrients, vitamins and minerals in the aetiology of squamous-cell carcinoma of the oesophagus. AB - Between 1992 and 1997 we conducted a case-control study of oesophageal cancer in 3 areas of northern Italy. Cases were 304 patients (29 women), ages 39-77 years (median age 60 years), with a first incident squamous-cell carcinoma (SCC) of the oesophagus. Controls were 743 patients (150 women), ages 35-77 years (median age 60 years), admitted for acute illnesses, unrelated to tobacco and alcohol, to major hospitals of the areas under surveillance. We derived estimates of daily dietary intake of 6 macronutrients, cholesterol, and 20 micronutrients or minerals from a validated food-frequency questionnaire, including 78 food groups and recipes and 15 questions on individual eating patterns. After allowance for age, gender, area of residence, education, body mass index, physical activity, smoking habit, alcohol consumption and energy intake, most micronutrients were inversely associated with oesophageal SCC risk. Highly significant associations emerged for monounsaturated fatty acids [odds ratio (OR) in highest vs. lowest intake quintile = 0.5]; carotene (OR = 0.3); lutein + zeaxanthin (OR = 0.4); vitamin C (OR = 0.4); and niacin (OR = 0.5). Only retinol appeared to be positively related to risk (OR = 1.9). The effect of the above nutrients, expressed as ORs, appeared to be similar in non-smokers and smokers, and non/light drinkers and heavy drinkers. PMID- 10797283 TI - Transcriptional inactivation of the tissue inhibitor of metalloproteinase-3 gene by dna hypermethylation of the 5'-CpG island in human gastric cancer cell lines. AB - The tissue inhibitor of metalloproteinase-3 (TIMP-3), a recently cloned member of TIMP gene family, has been implicated in the negative regulation of tumor cell invasion and tumor growth. Down-regulation of this gene has been shown to occur in a mouse carcinogenesis model, suggesting that it might play a role in the tumor progression of some cancers. In this study, we used human gastric cancer cell lines to investigate whether TIMP-3 gene expression is suppressed in human gastric cancer. We examined whether aberrant DNA methylation of the 5'-CpG island of the TIMP-3 gene is involved in this cancer. Nine of 10 human gastric cancer cell lines completely lost TIMP-3 gene expression compared with normal samples. Southern blot analysis and bisulfite genomic sequencing revealed aberrant hypermethylation near the transcription-start site of the TIMP-3 gene in all cell lines lacking TIMP-3 expression. Treatment of these cell lines with the demethylating agent 5-aza-2'-deoxycytidine restored TIMP-3 gene expression. Our results suggest that the TIMP-3 gene is another early target of tumor-associated aberrant DNA methylation in human gastric carcinogenesis. Consequently, genetic silencing of TIMP-3 may lead to a more malignant and invasive phenotype in these cancer cells. PMID- 10797284 TI - Lowering of tumoral interstitial fluid pressure by prostaglandin E(1) is paralleled by an increased uptake of (51)Cr-EDTA. AB - High intra-tumoral fluid pressure (TP(IF)) may impair uptake of anticancer drugs into tumors, contributing to poor efficiency in treatment of carcinomas. Here, we demonstrate that lowering of TP(IF) parallels increased transport of (51)Cr-EDTA (m.w. 341) into tumor interstitium. Introduction of 15 microg prostaglandin E(1) (PGE(1)) -methyl ester into the s.c. tissue surrounding transplanted rat colonic (PROb) carcinomas or chemically-induced rat mammary carcinomas, lowered TP(IF) by 30%. Transcapillary transport into the interstitium of PROb tumors quantified by microdialysis increased by 39.6% after PGE(1) treatment 40 min prior to administration of (51)Cr-EDTA (n=6; p<0.05) compared to vehicle (n=10). In mammary tumors, PGE(1) increased transport into the tumors by 86.9% over controls (n=16; p<0.05). Both tumors had well developed stroma containing collagen and hyaluronan. Our data demonstrate that adjuvant treatment with PGE(1) lowers TP(IF), and enhances transport into the tumors. This principle may be of value as adjuvant therapy in treatment of solid malignancies with currently used anticancer drugs. PMID- 10797285 TI - ERbB-2 expression is rate-limiting for epidermal growth factor-mediated stimulation of ovarian cancer cell proliferation. AB - Over-expression of the ErbB-2 proto-oncogene frequently coincides with an aggressive clinical course of certain human adenocarcinomas. The ErbB-2 receptor is a member of the ErbB family of growth factor receptors, and within this complex signaling network, ErbB-2-containing heterodimers are preferentially formed. To assess whether ErbB-2 is a critical component in epidermal growth factor (EGF)-mediated stimulation of tumor cell proliferation, we used as a model SK-OV-3 ovarian cancer cells, which over-express EGF receptor (EGFR) and ErbB-2 receptors. In these cells, we reduced ErbB-2 mRNA and protein expression by transfection with ErbB-2-targeted hammerhead ribozymes and generated cell lines expressing different levels of ErbB-2. In SK-OV-3 cells, ErbB-2 expression conferred a growth advantage and soft agar experiments revealed that ErbB-2 was rate-limiting for anchorage-independent growth. The induction of colony formation by EGF was completely abrogated in ErbB-2-depleted cells, despite unchanged expression levels and tyrosine phosphorylation of the EGFR. The duration of EGF mediated c-Fos mRNA up-regulation was decreased in parallel with loss of ErbB-2 expression. Furthermore, the rate of spontaneous apoptosis was increased in ErbB 2-depleted cells. Our results demonstrate that in human ovarian cancer cells the EGFR-ErbB-2 heterodimer, and not the EGFR homodimer, can be rate-limiting for EGF mediated proliferation, thus suggesting that the oncogenic activity of ErbB-2 in human tumors is due in part to its ability to increase the growth response to stroma-derived EGF-like growth factors. PMID- 10797286 TI - Expression of an activated Notch4(int-3) oncoprotein disrupts morphogenesis and induces an invasive phenotype in mammary epithelial cells in vitro. AB - The protein encoded by the Notch4 gene is a member of the Notch/lin-12 family of transmembrane receptor proteins, which have been shown to control cell fate determination and cell differentiation in a wide variety of organisms. Expression of Notch4(int-3), a truncated form of Notch4 having most of its extracellular domain deleted, as a transgene in mice induces the formation of poorly differentiated mammary carcinomas. To establish whether Notch4(int-3) has the capacity of subverting normal epithelial architecture, we assessed the effect of Notch4(int-3) expression on the in vitro morphogenetic properties of TAC-2 mammary epithelial cells. When grown in three-dimensional collagen gels in the presence of hydrocortisone, both wild-type and LacZ-transfected TAC-2 cells formed alveolar-like structures composed of polarized epithelial cells surrounding a central lumen. In contrast, TAC-2 cells programmed to express Notch4(int-3) formed compact cell aggregates devoid of tissue-specific organization. In addition, when grown on the surface of a collagen gel, Notch4(int-3)-expressing TAC-2 cells invaded the underlying matrix, whereas TAC-2 LacZ cells remained strictly confined to the gel surface. Expression of Notch4(int-3) in TAC-2 cells also disrupted contact-inhibition of cell proliferation, resulting in cell multilayering. Our results suggest that the ability of Notch4(int-3) to subvert normal epithelial morphogenesis and to promote invasion of the extracellular matrix contributes significantly to its tumorigenic potential. PMID- 10797287 TI - Expression of E-cadherin reduces bcl-2 expression and increases sensitivity to etoposide-induced apoptosis. AB - Expression of Bcl-2 is important in determining cancer cell resistance to chemotherapy. However, it is not clear whether cell-cell interactions regulate Bcl-2 expression. Using rat breast carcinoma cells selected for loss of hormone responsiveness, we found that parental E-cadherin-expressing cells (E cells) were more sensitive to etoposide-induced apoptosis than hormone-non-responsive cells (F cells), which failed to express E-cadherin. Expression of beta-catenin and pp120 src substrate proteins, which associate with E-cadherin, was unaffected. To determine whether re-expression of E-cadherin in F cells would restore etoposide sensitivity, F cells were transfected with an expression vector coding for the mouse E-cadherin gene. Stable clonal isolates expressing E-cadherin (F. Cad) showed increased sensitivity to etoposide treatment compared with control clones (F.Neo). Expression of E-cadherin resulted in a redistribution of beta-catenin from the cytoskeletal/nuclear fraction to the cytoplasmic/membrane fraction of the cells. E-cadherin-expressing clones also showed reduced invasion through basement membrane. Etoposide-induced apoptosis was characterized by morphological changes (nuclear blebbing) and DNA fragmentation. Induction of CPP32-like caspase activity was also observed in F.Cad transfectants but not F.Neo cells. Unlike F cells, F.Cad transfectants were not able to express Bcl-2, but transient transfection of bcl-2 resulted in re-expression and resistance to etoposide treatment. Therefore, E-cadherin may negatively regulate Bcl-2 expression by altering the availability of nuclear beta-catenin. Loss of E-cadherin in invasive tumor cells may lead to increased Bcl-2 expression and resistance to chemotherapeutic drugs. PMID- 10797288 TI - Enhanced expression of cyclooxygenase (COX)-2 in human skin epidermal cancer cells: evidence for growth suppression by inhibiting COX-2 expression. AB - Cyclooxygenase (COX)-2 is one of the rate-limiting enzymes in the conversion of arachidonic acid to prostaglandins and other eicosanoids. Recent studies have shown enhanced expression of COX-2 in cancer cells of several tissues. We investigated the expression of COX-2 and prostaglandin (PG) E((2)) production in two human skin epidermal cancer cell lines: cutaneous squamous cell carcinoma, HSC-5, and eccrine carcinoma, EcCa. Both COX-2 expression and PGE((2)) production were significantly enhanced in cancer cell lines compared with the non tumorigenic human keratinocyte cell line, HaCaT. In order to determine the role of COX-2 in the proliferation of HSC-5 and EcCa, the growth of untreated cells and cells transfected with COX-2 antisense oligonucleotide was compared using the MTT assay. Transfection with the antisense oligonucleotide suppressed COX-2 protein expression and significantly inhibited cell growth. The effect of a selective inhibitor of COX-2, NS398, was compared with the effect of the antisense oligonucleotide in order to see whether COX-2 expression and prostaglandins have selective effects on cell growth. COX-2 expression was unchanged by NS398 treatment, whereas NS398 inhibited cell growth to a certain extent. The degree of growth inhibition was greater with the antisense oligonucleotide than with NS398. Our findings indicate that COX-2 protein expression is enhanced in skin epidermal cancer cells and that COX-2 plays a pivotal role in regulating cell growth. Furthermore, inhibition of COX-2 expression had a more significant effect on growth suppression than inhibition of COX-2 catalytic activity, suggesting the existence of two different signal pathways via COX-2 in regulating cell growth. PMID- 10797289 TI - Genomic structure and mutation screening of the E2F4 gene in human tumors. AB - E2F4, a member of the E2F family of transcription factors, is abundant in non proliferating and differentiated cells where it plays an important role in the suppression of proliferation-associated genes. The E2F4 gene spans 6 kb and has 10 exons. It contains a serine (CAG) repeat tract in exon 7, which is unstable in gastrointestinal tumors. To further investigate a possible role of this gene in tumorigenesis we performed mutational analysis and expression studies in different tumors. Primary human tumor tissue of the stomach, colon, breast and lung (28), metastatic tumors of the colon (3) and small cell lung tumor cell lines (18) were screened for somatic mutations in the coding region of E2F4. No mutation was found. Microsatellite instability of the CAG repeat, however, was documented in primary stomach and colon tumors. Northern blot analysis revealed upregulated E2F4 transcript levels in tumor cell lines. Our data suggest that a direct involvement of E2F4 in tumorigenesis is unlikely, although increased E2F4 expression may be associated with human cancer. PMID- 10797290 TI - Synchronous multiple primary gastrointestinal cancer exhibits frequent microsatellite instability. AB - Colorectal (CRC) and gastric cancers (GC), the most common gastrointestinal malignancies, have been known to develop occasionally in a same patient. Previous studies have focused on the etiology of patients with multiple primary gastric and colorectal cancer (MPGCC); however, the carcinogenic process of MPGCC remains unclear. In this study, we have examined the genetic alterations in MPGCC in order to clarify the carcinogenic pathway. Twenty patients with sporadic MPGCC were examined for microsatellite instability (MSI) and frameshift mutations of target genes such as TGFbetaRII, BAX and IGFIIR. In 10 (50%) of 20 patients with MPGCC, MSI was present at least at 1 lesion of GC or CRC. Four (50%) of 8 cases with synchronous MPGCC displayed MSI in both GC and CRC, while only 1 (8%) of 12 cases of metachronous MPGCC exhibited MSI in both organs. Carcinogenic process of MPGCC was fairly associated with the MSI pathway, particularly in cases of synchronous MPGCC. MSI was found in 5 (25%) of 20 GCs and in 10 (50%) of 20 CRCs. MSI was involved more closely in CRC than in GC among MPGCC. Although most frameshift mutations at target genes were found in the MSI-positive MPGCC, infrequent mutations were observed in the genes. Frameshift mutation was found in only 1 of 5 cases of MSI-positive GC at TGFbetaRII. Only 2 of 10 cases of CRC with MSI showed mutation at TGFbetaRII, and 1 case also showed mutation at BAX and IGFIIR. Our findings suggest that TGFbetaRII, BAX and IGFIIR are not the main target genes for carcinogenesis in MSI-positive MPGCC. PMID- 10797291 TI - Frequent gain of the p40/p51/p63 gene locus in primary head and neck squamous cell carcinoma. AB - We have identified a new human p53 homologue, p40 (p51/p63). This gene was mapped to the distal arm of 3q and was found to be essential for normal epithelial development. We used microsatellite and FISH analyses to search for genetic alterations of p40 in primary HNSCC. A more precise localization of p40 was completed using 6 known markers on 3q and a newly isolated microsatellite marker within the p40 gene. We also determined the genomic organization of the p40 gene using human YAC and BAC clones. Microsatellite analysis revealed that 14 of 26 (54%) primary HNSCC had allelic imbalance in at least 1 of the 7 microsatellite loci. However, FISH analysis with a p40 probe showed that a majority of HNSCC had an increased copy number of the locus regardless of allelic status. Thus, overrepresentation of the p40 locus may play an important role in the development of HNSCC. PMID- 10797292 TI - ARHI is the center of allelic deletion on chromosome 1p31 in ovarian and breast cancers. AB - In our previous work, we had characterized ARHI as an imprinted putative tumor suppressor gene in ovarian and breast cancers. ARHI is expressed in primary breast and ovarian cell lines but largely absent from the corresponding malignant tumors. Moreover, the non-imprinted functional allele is typically deleted in malignant cells. Since ARHI had been mapped to 1p31, a common deletion site in breast and ovarian cancer and male germ-cell tumors, in this study, we set out to define precisely the physical location of ARHI at 1p31 and to determine if this location lies within the smallest common region of deletion in breast and ovarian cancers. To this end, we first carried out radiation hybrid mapping of ARHI and surrounding markers, followed by a high-resolution study of loss of heterozygosity at 1p31 in 49 ovarian and breast cancers. Combining a radiation hybrid map and a physical map of the region encompassing ARHI, 3 discrete regions of minimal deletion were found at 1p31 in breast and ovarian cancers. ARHI is the most common deletion region at 1p31. Two other less common regions of deletion were found centromeric to this gene. One of them centered on D1S207 and the other one included and was proximal to D1S488. We also confirmed the preferential loss of non-imprinted functional allele in 7 of 9 tumor specimens. These data support the possibility that ARHI is a tumor-suppressor gene and suggest that additional tumor-suppressor genes may lie proximal to ARHI at 1p31. The data obtained from our study should aid in the identification and characterization of genes in this novel imprinted region. PMID- 10797293 TI - Analysis of mutations in the URR and E6/E7 oncogenes of HPV 16 cervical cancer isolates from central China. AB - High rates of cervical cancer have been reported from parts of China and this may reflect a predominance of cervical infection with particularly aggressive human papillomavirus (HPV) variants. This PCR-based investigation of cervical tumours from Sichuan province in central China demonstrated an HPV positivity rate of 88%. HPV 16 was most common (21/34, 61%), followed by HPV 18 (3/34, 9%), while types 33, 45, 58 and 59 were each identified in one specimen. Sequencing of up to 1349 bases of the 21 HPV 16-positive isolates, encompassing the enhancer/promoter of the upstream regulatory region (URR) and the E6 and E7 genes, revealed distinct patterns of genomic stability and variability. An overall mutation rate of 5% was seen in the URR. One isolate had a large deletion of 436 bases in the enhancer; while varying combinations of 21 point mutations were identified in the remainder, impacting several YY1, NF1, TEF-1 and Oct-1 sites. More sequence variations were found in E6 compared to E7 (81% vs. 52% of isolates showing at least one mutation), some of which resulted in changes to the translated amino acids. Since the E6/E7 genes encode the oncogenic proteins essential for malignant transformation, and as their expression is controlled by the URR, it is possible that some of the identified mutations altered the oncogenicity of the virus: either directly by changing amino acid sequences of the E6 or E7 oncoproteins, or indirectly through alterations to transcription factor binding sites in the URR. PMID- 10797294 TI - Reactivity of natural and induced human antibodies to MUC1 mucin with MUC1 peptides and n-acetylgalactosamine (GalNAc) peptides. AB - Antibodies (Abs) to MUC1 occur naturally in both healthy subjects and cancer patients and can be induced by MUC1 peptide vaccination. We compared the specificity of natural and induced MUC1 Abs with the objective of defining an effective MUC1 vaccine for active immunotherapy of adenocarcinoma patients. Serum samples, selected out of a screened population of 492 subjects for their high levels of IgG and/or IgM MUC1 Abs, were obtained from 55 control subjects and from 26 breast cancer patients before primary treatment, as well as from 19 breast cancer patients immunized with MUC1 peptides coupled to keyhole limpet hemocyanin (KLH) and mixed with QS-21. The samples were tested with enzyme-linked immunoassays for reactivity with (1) overlapping hepta- and 20-mer peptides spanning the MUC1 tandem repeat sequence; (2) two modified 60-mer peptides with substitutions in the PDTR (PDTA) or in the STAPPA (STAAAA) sequence of each tandem repeat; and (3) four 60-mer glycopeptides with each 1, 2, 3 and 5 mol N acetylgalactosamine (GalNAc) per repeat. More than one minimal epitopic sequence could be defined, indicating that Abs directed to more than one region of the MUC1 peptide core can coexist in one and the same subject. The most frequent minimal epitopic sequence of natural MUC1 IgG and IgM Abs was RPAPGS, followed by PPAHGVT and PDTRP. MUC1 peptide vaccination induced high titers of IgM and IgG Abs predominantly directed, respectively, to the PDTRPAP and the STAPPAHGV sequences of the tandem repeat. Natural MUC1 Abs from breast cancer patients reacted more strongly with the N-acetylgalactosamine (GalNAc) peptides than with the naked 60-mer peptide, while reactivity with the GalNAc-peptides was significantly reduced (2-tailed p < 0.0001) in the MUC1 IgG and IgM Abs induced by MUC1 peptide vaccination. Whereas in cancer patients glycans appear to participate in epitope conformation, the epitope(s) recognized by MUC1 Abs induced by peptide vaccination are already masked by minimal glycosylation. Therefore, our results indicate that a MUC1 glycopeptide would be a better vaccine than a naked peptide. PMID- 10797295 TI - Susceptibility of rat colon carcinoma cells to lymphokine activated killer mediated cytotoxicity is decreased by alpha1,2-fucosylation. AB - The presence of alpha1,2-fucosylated glycans at the surface of rat colon carcinoma cells has been associated with an increased tumorigenicity and resistance to natural killer/lymphokine activated killer (NK/LAK) cytotoxicity. We now report that transfection of rat alpha1,2-fucosyltransferases cDNA (FTA and FTB) into REG cells, which are spontaneously devoid of this enzymatic activity, allows expression of histo-blood group H antigen and increases their resistance to LAK, but not NK cell lysis. Conversely, transfection of PRO cells, which spontaneously express alpha1, 2-fucosyltransferase activity, with the FTA cDNA in the antisense orientation decreases expression of the H antigen together with their resistance to LAK cell lysis, but again, not to NK cell lysis. Furthermore, REG cells that are rejected by immunocompetent syngeneic rats are similarly rejected by rats depleted of NK cells by antibody 3.2.3, directed against the NKR P1 molecule. Thus, the rejection of REG cells by immunocompetent rats and their earlier reported increased tumorigenicity after transfection with an alpha1, 2 fucosyltransferase cDNA cannot be ascribed to NK cell sensitivity or resistance, respectively. The increased resistance to LAK cell lysis, however, may be relevant to tumor progression. PMID- 10797296 TI - Recombinant anti-CD25 immunotoxin RFT5(SCFV)-ETA' demonstrates successful elimination of disseminated human Hodgkin lymphoma in SCID mice. AB - Since clinical phase-I/II trials in patients with resistant Hodgkin's lymphoma treated with the chemically linked anti-CD25 ricin-A-chain immunotoxin RFT5-SMPT dgA indicate promising results for patients with minimal residual disease, we constructed a new immunotoxin by fusing the RFT5 single-chain variable fragment to a deletion mutant of Pseudomonas exotoxin A (ETA'). The recombinant protein was directed into the periplasmic space of E. coli by means of the pET-derived expression vector pBM1.1 and our newly developed expression/purification method. Biologically active RFT5(scFv)-ETA' was isolated by freezing/thawing and purified by immobilized metal-ion affinity and molecular-size-chromatography. RFT5(scFv) ETA' was subsequently used for the treatment of disseminated human Hodgkin's lymphoma in a SCID-mouse model. The mean survival time (MST) of L540rec challenged SCID mice was 38.1 days. A single i.v. injection of 40 microg recombinant immunotoxin (rIT) 1 day after tumor inoculation resulted in 100% tumor-free mice, extending the MST to more than 220 days (p < 0.0001). The blood distribution time T(1/2)alpha was 39.65 min, the serum elimination time T(1/2)alpha, 756.6 min. All animals were assessed for soluble interleukin-2 receptor alpha, which is directly correlated to tumor burden. Soluble CD25 was not detectable in mice treated with the rIT. Our findings, concerning potent anti tumor effects of a recombinant anti-CD25 immunotoxin against disseminated Hodgkin's lymphoma in SCID mice reported here demonstrate that RFT5(scFv)-ETA' might be suitable for further evaluation against Hodgkin's lymphoma in humans. PMID- 10797297 TI - Antigen-specific cancer immunotherapy using a GM-CSF secreting allogeneic tumor cell-based vaccine. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF)-transduced autologous tumor cell-based vaccines are currently one of the major forms of cancer vaccines. However, the preparation of GM-CSF-transduced autologous tumor vaccines is time-consuming and technically challenging. In addition, the host antigen presenting cells, rather than the tumor vaccine cells themselves, present tumor specific antigens and prime the host T cells. Therefore, we tested the efficacy of antigen-specific allogeneic tumor vaccines. We used human papillomavirus 16 (HPV-16) E7 protein as a model tumor antigen, which is associated with the development of most cervical carcinoma. B16, a C57BL/6 (H-2(b)) derived melanoma cell line, was genetically engineered to produce GM-CSF alone (B16GM), HPV-16 E7 alone (B16E7), or both (B16GME7). These vaccine cells were injected into BALB/c (H-2(d)) mice (10(6) cells/mouse). Two weeks later, mice were challenged with 10(5) live HPV-16 E7(+) BL-1 (H-2(d)) tumor cells and monitored for tumor progression twice weekly. To determine the effective cell population in the antitumor immunity elicited by B16GME7, we carried out in vivo antibody depletion experiments using CD4 and CD8 specific antibodies. In addition, as a measure of the immune responses produced by B16GME7, we performed an in vitro cytotoxic T lymphocyte assay using a standard chromium release method. We found that all of the mice vaccinated with B16GME7 remained tumor free 49 days post-BL-1 challenge. In contrast, mice vaccinated with B16GM and B16E7 did not show any tumor protection against a similar dose of BL-1 cells. Furthermore, the antitumor immunity produced by B16GME7 was dependent on both CD4 and CD8 T cells. In addition, E7-specific cytotoxic T lymphocyte activity could be readily demonstrated in mice immunized with B16GME7. These results suggest that allogeneic tumor cells transduced with GM-CSF and the tumor antigen, HPV-16 E7, cannot only generate an E7-specific cytotoxic T lymphocytes response in vitro, but can also elicit a potent antitumor immune response against an E7 expressing tumor in vivo. PMID- 10797298 TI - Classification of human ovarian tumors using multivariate data analysis of polypeptide expression patterns. AB - Large amounts of data on quantitative gene expression are generated by procedures such as 2-DE analysis of proteins or cDNA microarrays. Quantitative molecular variation may potentially be used for the development of methods for the classification of tumors. We used here the statistical concepts of principal components analysis (PCA) and partial least square analysis (PLS) in an attempt to type ovarian tumors. Using a set of 170 polypeptides, 22 tumors were used to establish a model ("learning set") for classification into 3 groups (benign/borderline/malignant). Eighteen tumors were then used to test the model. Six of 8 carcinomas and 3 of 4 borderline tumors were correctly classified. Two of 6 benign lesions were correctly classified, 3 were classified as borderline and 1 as carcinoma. We conclude that it may be possible to classify tumors according to their constitutive protein expression profile using multivariate analysis, thus making classification by artificial intelligence a future possibility. PMID- 10797299 TI - Prevalence of founder BRCA1 and BRCA2 mutations among breast and ovarian cancer patients in Hungary. AB - We have investigated the impact of BRCA1 and BRCA2 mutations that were frequently identified among Hungarian high-risk breast-ovarian cancer families (Ramus et al., 1997b, AJHG), on the development of breast and ovarian cancer in the general Hungarian population. The prevalence of 3 BRCA1 mutations (185delAG, 300T-->G and 5382insC) and 2 BRCA2 mutations (6174delT and 9326insA) was evaluated in a hospital-based consecutive series of 500 female breast cancer patients and 90 ovarian cancer patients, not selected for age at diagnosis or family history of cancer, as well as in 350 controls. Among breast cancer patients, 3.6% (18/500) carried a founder mutation: 9 BRCA1 300T-->G, 7 BRCA1 5382insC, 1 BRCA1 185delAG and 1 BRCA2 9326insA. Among ovarian cancer patients, 11% (10/90) carried a founder mutation: 5 BRCA1 185delAG, 4 BRCA1 300T-->G and 1 BRCA1 5382insC. One control carried a mutation, BRCA1 5382insC. Inherited breast cancer was more frequent among women with younger age at diagnosis: 6.1% of women younger than age 50 but 2.4% of women diagnosed at age 50 or older carried one of the founder mutations. There was no association between mutation status and age at diagnosis of ovarian cancer. Three of 23 medullary breast cancers were inherited (p = 0.038). Carrier status was also associated with a non-significant trend toward advanced tumor stage at diagnosis. These mutations could be evaluated among all ovarian cancer patients and breast cancer patients younger than age 60 and of Hungarian ancestry. PMID- 10797300 TI - Applicability of an absolute quantitative procedure to monitor intra-individual bcr/abl transcript kinetics in clinical samples from chronic myelogenous leukemia patients. AB - Chimeric bcr/abl fusion proteins are thought to be the molecular 'pathogen' of chronic myelogenous leukaemia (CML). Expression levels of the respective fusion RNAs reflect disease progression as well as remission upon therapeutic intervention in CML patients. However, there is no quick and reliable method that would allow the quantitative routine monitoring of bcr/abl hybrid transcripts. A fluorescent probe-based PCR assay (TaqMan) has been described to quantitfy the exact amount of target sequences. We have established TaqMan real-time RT-PCRs for M-bcr/abl (b2a2, b2a3, b3a2, b3a3) and m-bcr/abl (e1a2) fusion transcripts as well as for beta-actin. All PCRs quantified as little as 10 copies/100 ng total cDNA. In order to investigate whether this procedure is appropriate for routine diagnostic monitoring, we performed retrospective measurements on 9 documented CML disease courses. Our data show that ongoing or relapsing CML is paralleled by increasing peripheral levels of bcr/abl fusion RNAs. Furthermore, sucessful anti leukemic treatment is reflected by decreasing absolute bcr/abl transcript numbers. In contrast with conventional bcr/abl PCR techniques we could distinguish single positive values and gradually increasing copy numbers. PMID- 10797301 TI - Perceived risk of cancer in population samples from 5 european countries. PMID- 10797302 TI - Anti-MAGE-3 antibody 57B and anti-MAGE-1 antibody 6C1 can be used to study different proteins of the MAGE-A family. PMID- 10797303 TI - Stretch-activated signaling of nerve growth factor secretion in bladder and vascular smooth muscle cells from hypertensive and hyperactive rats. AB - Elevated vascular (VSMC) and bladder smooth muscle (BSMC) NGF are associated with altered visceral innervation in the spontaneously hypertensive rat (SHR: hypertensive, behaviorally hyperactive) compared with control Wistar-Kyotos (WKYs). Stretch stimulates increased NGF production in BSMCs. To elucidate whether stretch induces NGF synthesis in VSMCs, and to determine if disturbances in stretch-mediated NGF production contribute to the elevated tissue levels of NGF in SHRs, we subjected VSMCs and BSMCs cultured from four established inbred rat strains (WKY, WKHA: hyperactive; SHR and WKHT: hypertensive) to several stretch paradigms. For VSMCs, acute and cyclic stretch affected cells derived from hypertensive rats (80-100% increase over control) but not from normotensive strains. For BSMCs, cyclic and static stretch increased NGF secretion in all four strains, but had a two- to threefold greater effect in cells from SHRs and WKHTs (increase up to 600%) at early time points. At later time points of a 24-h experimental period, stretch increased NGF output up to 400% in SHR and WKHA cultures. Thus, defects that influence early induction of stretch-mediated SHR NGF secretion cosegregate with the hypertensive phenotype. Stretch-gated ion channel inhibitors, voltage-gated ion channel inhibitors, and protease inhibitors failed to affect stretch-induced BSMC NGF secretion. In contrast, gene transcription, intracellular calcium, protein kinase C (PKC), and autocrine release of an unknown factor may play a role in the elevated NGF secretion observed in smooth muscle from hypertensive animals. Altered stretch-induced smooth muscle NGF secretion may contribute to the augmented vascular and bladder NGF content associated with high blood pressure and hyperactive voiding in SHRs. PMID- 10797304 TI - Heregulin-beta is especially potent in activating phosphatidylinositol 3-kinase in nontransformed human mammary epithelial cells. AB - The neu differentiation factors/heregulins (HRGs) comprise a family of polypeptide growth factors that activate p185(erbB-2) through direct binding to either erbB-3 or erbB-4 receptor tyrosine kinases. We have previously shown that HRG-beta is mitogenic for various human mammary epithelial cell lines that coexpress c-erbB-2 and c-erbB-3. Phosphatidylinositol 3-kinase (PI3K) is activated by p185(erbB-2) /erbB-3 heterodimers in cells stimulated by HRG, and PI3K is constitutively activated by p185(erbB-2) /erbB-3 in breast carcinoma cells that overexpress c-erbB-2. To better understand the relative abilities of HRGs, epidermal growth factor (EGF), or insulin to activate PI3K under normal physiological conditions, we compared the levels of recruitment of the 85-kDa regulatory subunit of PI3K when activated by the type I (erbB) or type II [insulin-like growth factor (IGF)] receptor tyrosine kinases in two different nontransformed human mammary epithelial cell lines. The nontransformed H16N-2 cells isolated from normal tissue express EGFR, p185(erbB-2), and erbB-3, and are highly responsive to the mitogenic effects of HRG-beta as well as to the combination of EGF and insulin in serum-free culture. We measured the stoichiometry of p85 recruited by tyrosine-phosphorylated proteins induced in H16N-2 cells by either the alpha or the beta isoform of HRG. HRG-beta was greater than 10-fold more potent in inducing p85 recruitment than was the less biologically active HRG-alpha isoform. HRG-beta was also a more potent inducer of p85 recruited by tyrosine-phosphorylated proteins than was either EGF, insulin, or EGF and insulin combined. Furthermore, erbB-3 principally mediated the direct recruitment of p85 in cells stimulated by HRG or EGF, indicating that, in addition to the high-level activation of PI3K by p185(erbB-2) / erbB-3, EGFR/erbB 3 heterodimer interaction is essential for the weak but significant level of PI3K activated by EGF in cells that express normal EGFR levels. Studies using the PI3K inhibitor wortmannin also indicated that PI3K activation was required for the proliferation of H16N-2 cells induced by either HRG-beta or EGF and insulin in serum-free culture. Finally, HRG-beta was also an especially potent inducer of PI3K in the nontransformed MCF-10A cells, which were derived spontaneously from normal reduction mammoplasty tissue. These data show, for the first time, a side by-side quantitative comparison of the relative degree of PI3K activated by different growth factors in nontransformed growth factor-dependent cells under precisely defined conditions in culture. PMID- 10797305 TI - Thrombin-stimulated phosphatidylinositol 3-kinase activity in platelets is associated with activation of PYK2 tyrosine kinase: activation of both enzymes is aggregation independent. AB - In this study, we investigated the activation of a new member of the focal adhesion kinase family of tyrosine kinases, the proline-rich tyrosine kinase, or PYK2, in platelets. We show that PYK2 is tyrosine phosphorylated and its activity is increased during early stages of platelet aggregation. This activation coincided with increased association of phosphatidylinositol (PI) 3-kinase and PYK2, as determined by both anti-PI 3-kinase and anti-PYK2 immunoprecipitates. However, under basal conditions, some association of PYK2 and PI 3-kinase was consistently observed, even though little or no tyrosine phosphorylated PYK2 could be detected. In addition, both increased PI 3-kinase activity and increased PYK2 activity could be detected in immunoprecipitates following thrombin stimulation. All of these events were unaffected by blocking platelet aggregation with arginine-glycine-aspartate-serine (RGDS) peptide, which interferes with binding of the platelet integrin alpha(IIb)beta(3) to fibrinogen. Neither was the activation of the PYK2 kinase activity affected by blocking PI 3-kinase activity. These results support a model in which PYK2 is associated with PI 3-kinase in unstimulated platelets and following activation of platelets, there is an increase in tyrosine phosphorylation of PYK2, increased PYK2 activity, and increased association of PYK2 with PI 3-kinase, which may contribute to the increase in PI 3-kinase activity. All of these were found to be early events independent of subsequent platelet aggregation. PMID- 10797306 TI - Ectopic expression of the ErbB-3 binding protein ebp1 inhibits growth and induces differentiation of human breast cancer cell lines. AB - Ebp1, an ErbB-3 binding protein, translocates from the cytoplasm to the nucleus of human breast cancer cells after treatment with the ErbB-3 ligand, heregulin. The purpose of these studies was to examine the effects of ectopic expression of ebp1 on the biological properties of human ErbB-3-expressing breast carcinoma cell lines. Ectopic expression of ebp1 in ErbB-2, ErbB-3-expressing breast carcinoma cell lines resulted in inhibition of colony formation, a decreased proliferation rate, an accumulation of cells in the G2/M phase of the cell cycle, and suppression of growth in soft agar. Ectopic expression of ebp1 led to a more differentiated phenotype in AU565 breast cancer cells, as evidenced by increased expression of lipid droplets and of the milk protein casein. Basal phosphorylation of extracellular regulated kinases (Erks) 1 and 2, kinases activated by heregulin treatment, was also observed in ebp1 transfectants. The promoter for the intercellular adhesion molecule-1 gene, a heregulin-inducible gene, was constitutively activated in ebp1 transfectants as determined by reporter construct analysis. These data demonstrate that ectopic expression of the ErbB-3 binding protein Ebp1 inhibits proliferation and induces differentiation of ErbB-2, ErbB-3-expressing human breast carcinoma cell lines. PMID- 10797307 TI - Tumor necrosis factor-alpha-induced apoptosis is associated with suppression of insulin-like growth factor binding protein-5 secretion in differentiating murine skeletal myoblasts. AB - Wasting of muscle and fat during cachexia exceeds that explained by reduced food intake alone. This wasting may result from an imbalanced cytokine environment, which could lead to increased protein catabolism. Supporting this, tumor necrosis factor-alpha (TNF-alpha) is raised in several animal models of cachectic muscle wasting. Therefore, we assessed the effects of TNF-alpha and its second messenger, ceramide, on the proliferation, differentiation, and survival of murine C2 skeletal myoblasts. Because insulin-like growth factor binding protein 5 (IGFBP-5) and insulin-like growth factor-II (IGF-II) are potent regulators of myoblast proliferation and differentiation, we monitored the ability of exogenous TNF-alpha to manipulate this system. Fibroblast growth factor (FGF) ceramide, or TNF-alpha suppressed differentiation of C2 cells compared with controls. All treatments suppressed IGF-II production but only TNF-alpha blocked IGFBP-5 secretion. TNF-alpha increased apoptotic cell death, which otherwise remained basal (low serum differentiation medium (LSM), FGF) or low (ceramide). Suppression of both IGFBP-5 and IGF-II secretion may explain why of all triggers tested, only TNF-alpha not only blocked differentiation, but also promoted cell death. This suggests a fundamental role of IGFBP-5 for maintaining muscle survival. Supporting this hypothesis, no increase in apoptosis was seen in IGFBP 5 cDNA tranfected C2 cells after TNF-alpha treatment. In summary, the IGF system is essential for maintaining skeletal muscle cell survival and differentiation, and its suppression by TNF-alpha is fundamental regarding muscle wasting, and may be associated in vivo with cancer cachexia. PMID- 10797308 TI - Involvement of hepatocyte epidermal growth factor receptor mediated activation of mitogen-activated protein kinase signaling pathways in response to growth inhibition by a novel K vitamin. AB - Compound 5 (Cpd 5), a synthetic K vitamin analogue, or 2-(2-mercaptoethanol)-3 methyl-1,4-naphthoquinone, is a potent inhibitor of epidermal growth factor (EGF) induced rat hepatocyte DNA synthesis and induces EGF receptor (EGFR) tyrosine phosphorylation. To understand the cellular responses to Cpd 5, its effects on the EGF signal transduction pathway were examined and compared to those of the stimulant, EGF. Cpd 5 induced a cellular response program that included the induction of EGFR tyrosine phosphorylation and the activation of the mitogen activated protein kinase (MAPK) cascade. EGFR tyrosine phosphorylation was induced by Cpd 5 in a time- and dose-dependent manner. Coimmunoprecipitation studies demonstrated that both EGF and Cpd 5 induced tyrosine phosphorylation of EGFR was associated with increased amounts of adapter proteins Shc and Grb2, and the Ras GTP-GDP exchange protein Sos, indicating the formation of functional EGFR complexes. Although EGFR phosphorylation was induced both by the stimulant EGF and the inhibitor Cpd 5, the timing and intensity of activation by EGF and Cpd 5 were different. EGF activated EGFR transiently, whereas Cpd 5 induced an intense and sustained activation. Cpd 5-altered cells had a decreased ability to dephosphorylate tyrosine phosphorylated EGFR, providing evidence for an inhibition of tyrosine phosphatase activity. Both EGF and Cpd 5 caused an induction of phospho-extracellular response kinase (ERK), which was also more sustained with Cpd 5. Moreover, whereas Cpd 5 induced a striking translocation of phosphorylated ERK from cytosol to the nucleus, no significant nuclear translocation occurred after stimulation with EGF. The data suggest that this novel compound causes growth inhibition through antagonism of EGFR phosphatases and consequent induction of EGFR and ERK phosphorylation. This is supported by experiments with PD 153035 and PD 098059, antagonists of phosphorylation of EGFR and MAP kinase kinase (MEK), respectively, which both antagonized Cpd 5-induced phosphorylation and the inhibition of DNA synthesis. These results imply a mechanism of cell growth inhibition associated with intense and prolonged protein tyrosine phosphorylation. Protein tyrosine phosphatases may thus be a novel target for drugs designed to inhibit cell growth. PMID- 10797309 TI - Divergent phenotypic patterns and commitment to apoptosis of Caco-2 cells during spontaneous and butyrate-induced differentiation. AB - Caco-2 cells differentiate spontaneously when cultured in confluence and on exposure to the physiologically relevant short-chain fatty acid, butyrate. This study aimed to compare the phenotype induced by these pathways and their relations to cell turnover. Caco-2 cells were treated with butyrate at a nontoxic concentration of 2 mM for 3 days, or allowed to spontaneously differentiate for 0 21 days. Brush border hydrolase activities and carcinoembryonic antigen (CEA) expression, transepithelial resistance and dome formation, expression of components of the urokinase system, and cell turnover by flow cytometry, and the degree of DNA fragmentation were quantified. Butyrate induced increases in alkaline phosphatase activity and CEA expression but not the activities of other hydrolases, while culture alone induced progressive increases in the activities/expression of all markers. Butyrate induced a significantly greater increase in transepithelial resistance (TER) than occurred during culture alone but the densities of domes were similar. Butyrate induced a ninefold increase in urokinase receptor expression and twofold increase in urokinase activity, while culture alone induced a significantly smaller increase in receptor expression, an increase in plasminogen activator inhibitor-1 but no change in activity. While both stimuli induced cell cycle arrest, only butyrate increased the proportion of cells undergoing apoptosis. In conclusion, differentiation of Caco-2 cells can proceed along multiple pathways but does not necessarily lead to apoptosis. The phenotypic changes during spontaneous differentiation mimic those that occur in normal colonic epithelial cells in vivo during their migration from the crypt base to neck, while butyrate-induced effects more closely follow those occurring when normal colonic epithelial cells migrate from crypt neck to the surface compartment. PMID- 10797310 TI - High glucose-induced inhibition of alpha-methyl-D-glucopyranoside uptake is mediated by protein kinase C-dependent activation of arachidonic acid release in primary cultured rabbit renal proximal tubule cells. AB - Abnormal glucose handling in the proximal tubule may play an important role in the development of diabetic nephropathy. Thus, the present study was designed to examine the effect of high glucose on alpha-methyl-D-glucopyranoside (alpha-MG) uptake and its signaling pathways in the primary cultured rabbit renal proximal tubule cells (PTCs). When PTCs were preincubated with 25 or 50 mM glucose for 4 h, 25 or 50 mM glucose significantly inhibited alpha-MG uptake, while 25 or 50 mM mannitol and L-glucose did not affect. Actinomycin D and cycloheximide did not block the effect of high glucose on alpha-MG uptake. Twenty-five millimoles glucose-induced inhibition of alpha-MG uptake was blocked by mepacrine and AACOCF(3), phospholipase A(2) (PLA(2)) inhibitors. Twenty-five millimoles of glucose, not mannitol or L-glucose, significantly increased the [(3)H] arachidonic acid (AA) release compared to control. In addition, the 25 mM glucose induced [(3)H]-AA release was completely blocked by mepacrine or AACOCF(3). Indomethacin, a cyclooxygenase inhibitor, blocked the high glucose-induced inhibition of alpha-MG uptake, although econazole, cytochrome P-450 a epoxygenase inhibitor, and nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor, did not. On the other hand, staurosporine and bisindolylmaleimide I, protein kinase C (PKC) inhibitors, blocked 25 mM glucose-induced increase of [(3)H]-AA release and inhibition of alpha-MG uptake. However, neomycin, U 73122, and phospholipase c(PLC) inhibitors did not block the effect of 25 mM glucose on [(3)H]-AA release and alpha-MG uptake. Pretreatment of methoxyverapamil, an L-type Ca(2+) channel blocker, abolished 25 mM glucose-induced increase of [(3)H]-AA release. Indeed, 25 mM glucose increased translocation of cPLA(2) from cytosolic fraction to membrane fraction. In conclusion, the present results demonstrate that high glucose inhibits alpha-MG uptake by the increase of AA release via the activation of PKC. PMID- 10797311 TI - Expression of functional mGlu5 metabotropic glutamate receptors in human melanocytes. AB - Cultured human melanocytes express mGlu5 metabotropic glutamate (mGlu) receptors, as shown by RT-PCR, immunocytochemistry, Western blot analysis, and measurement of agonist-stimulated polyphosphoinositide hydrolysis. The mGlu5 receptor agonists (S)-3, 5-dihydroxyphenylglycine and quisqualate increased [(3)H methyl]thymidine incorporation and melanocyte proliferation in subconfluent cultures, but impaired cell viability in confluent cultures. Both effects were prevented by 2-methyl-6-(2-phenyl-1-ethynyl)-pyridine, a potent and highly selective mGlu5 receptor antagonist. Agonists of other mGlu receptor subtypes (such as the mGlu2/3 receptor agonist, 2S,2'R,3'R-2-2', 3' dicarboxycyclopropylglycine, or the mGlu4/6/7/8 receptor agonist, L-2-amino-4 phosphonobutanoate) or selective agonists of ionotropic glutamate receptors (N methyl-D-aspartate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate, and kainate) did not affect melanocyte proliferation or viability. The presence of a receptor for glutamate, the major excitatory neurotransmitter, in human melanocytes is intriguing. mGlu5 receptors may be involved in the control of melanocyte proliferation (and perhaps in other functions), but harbor a potential toxicity and may therefore contribute to cell damage under pathological conditions. PMID- 10797312 TI - Intracellular pH regulation in a nonmalignant and a derived malignant human breast cell line. AB - Tumor cells in vivo often exist in an ischemic microenvironment that would compromise the growth of normal cells. To minimize intracellular acidification under these conditions, these cells are thought to upregulate H(+) transport mechanisms and/or slow the rate at which metabolic processes generate intracellular protons. Proton extrusion has been compared under identical conditions in two closely related human breast cell lines: nonmalignant but immortalized HMT-3522/S1 and malignant HMT-3522/T4-2 cells derived from them. Only the latter were capable of tumor formation in host animals or long-term growth in a low-pH medium designed to mimic conditions in many solid tumors. However, detailed study of the dynamics of proton extrusion in the two cell lines revealed no significant differences. Thus, even though the ability to upregulate proton extrusion in a low pH environment (pH(e)) may be important for cell survival in a tumor, this ability is not acquired along with the capacity to form solid tumors and is not unique to the transformed cell. This conclusion was based on fluorescence measurements of intracellular pH (pH(i)) on cells that were plated on extracellular matrix, allowing them to remain adherent to proteins to which they had become attached 24 to 48 h earlier. Proton translocation under conditions of low pH(e) was observed by monitoring pH(i) after exposing cells to an acute acidification of the surrounding medium. Proton translocation at normal pH(e) was measured by monitoring the recovery after introduction of an intracellular proton load by treatment with ammonium chloride. Even in the presence of inhibitors of the three major mechanisms of proton translocation (sodium-proton antiport, bicarbonate transport, and proton-lactate symport) together with acidification of their medium, cells showed only about 0.4 units of reduction in pH(i). This was attributed to a slowing of metabolic proton generation because the inhibitors were shown to be effective when the same cells were given an intracellular acidification. PMID- 10797313 TI - Interaction of smad3 with a proximal smad-binding element of the human alpha2(I) procollagen gene promoter required for transcriptional activation by TGF-beta. AB - Transcription of the alpha2(I) collagen gene (COL1A2) in fibroblasts is potently induced by transforming growth factor-beta (TGF-beta). Smad family proteins function as intracellular signal transducers for TGF-beta that convey information from the cell membrane to the nucleus. In the present study, we establish the functional requirement for endogenous Smad3 and Smad4 in TGF-beta-stimulated COL1A2 transcription in human skin fibroblasts in vitro. Furthermore, using transfections with a series of 5' deletions of the human COL1A2 promoter, we identify a proximal region between -353 and -148 bp, which is required for full stimulation of transcription by a constitutively active TGF-beta type I receptor. This region of the COL1A2 promoter contains a CAGA motif also found in the promoter of the plasminogen activator inhibitor-1. Substitutions disrupting this sequence decreased the binding of nuclear extracts or recombinant Smad3 to the CAGACA oligonucleotide, and markedly reduced the transcriptional response to TGF beta or overexpressed Smad3 in transient transfection assays. The insertion of tandem repeats of CAGACA conferred TGF-beta stimulation to a heterologous minimal promoter-reporter construct. Inhibition of endogenous Smad expression in fibroblasts by antisense oligonucleotides or cDNA against Smad3 or Smad4, and transfection of COL1A2 promoter constructs into Smad4-deficient breast adenocarcinoma cells, indicated the critical role of Smads for the full TGF-beta response. The importance of Smad binding to the CAGACA box of COL1A2 was further established by transcriptional decoy oligonucleotide competition. Taken together, the results identify a functional Smad-binding element of the COL1A2 promoter harboring a CAGACA consensus sequence that is both necessary and sufficient for stimulation by TGF-beta, and demonstrate that interaction of this Smad-binding element with endogenous Smads is required for the full TGF-beta response in fibroblasts. PMID- 10797314 TI - Adenosine acts as a chemoprotective agent by stimulating G-CSF production: a role for A1 and A3 adenosine receptors. AB - Adenosine, a ubiquitous nucleoside, is released into the extracellular environment from metabolically active or stressed cells. It binds to cells through specific A1, A(2A), A(2B), and A3 G-protein-associated cell-surface receptors, thus acting as a signal-transduction molecule by regulating the levels of adenylyl cyclase and phospholipase C. In this study, we showed that adenosine stimulates the proliferation of murine bone marrow cells in vitro. Pharmacological studies, using antagonists to the adenosine receptors, revealed that this activity was mediated through the binding of adenosine to its A1 and A3 receptors. This result was further corroborated by showing that the two selective A1 and A3 receptor agonists, N-cyclopentyladenosine (CPA) and 1-deoxy-1-[6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-be ta-D-ribofuranuronamide (IB MECA) respectively, induced bone marrow cell proliferation in a manner similar to adenosine. Adenosine's interaction with its A1 and A3 receptors induced G-CSF production, which led to its stimulatory effect on bone marrow cells. These results were confirmed in vivo when we demonstrated that low-dose adenosine (0.25 mg/kg) acted as a chemoprotective agent. When administered after chemotherapy, it restored the number of leukocytes and neutrophils to normal levels, compared with the decline in these parameters after chemotherapy alone. It is suggested that low-dose adenosine, already in clinical use, may also be applied as a chemoprotective agent. PMID- 10797316 TI - Grb7 signal transduction protein mediates metastatic progression of esophageal carcinoma. AB - We have previously reported the association of tumor cell invasion with expression of growth factor receptor-bound protein 7 (Grb7). This molecule contains a Src homology 2 (SH2) domain and shares structural homology with a cell migration molecule designated Mig-10 found in Caenorhabditis elegans. In the present study, Grb7 expression was analyzed in human esophageal carcinomas with or without metastatic spread. The Grb7 protein was overexpressed in 14 of 31 esophageal carcinomas as compared to the adjacent normal mucosa (45%) and this finding was significantly correlated with the presence of lymph node metastases. We also identified that Grb7 protein in esophageal carcinoma cells was phosphorylated on tyrosine by epidermal growth factor as well as attachment to extracellular matrix proteins including fibronectin. Such fibronectin-dependent phosphorylation of Grb7 was regulated by integrin signaling that leads to the interaction with focal adhesion kinase protein. Furthermore, ectopic expression of a Grb7-SH2 dominant-negative fragment inhibited the fibronectin-dependent phosphorylation of endogenous Grb7, and reduced migration of esophageal carcinoma cells into fibronectin. Our results suggest a role of Grb7 mediated signal transduction in generation of an invasive cell phenotype against extracellular matrix, and thus contributes to metastatic progression of human esophageal carcinoma. PMID- 10797315 TI - Analysis of Myc/Max/Mad network members in adipogenesis: inhibition of the proliferative burst and differentiation by ectopically expressed Mad1. AB - Transcription factors of the Myc/Max/Mad network affect multiple aspects of cellular behavior, including proliferation, differentiation, and apoptosis. Recent studies have shown that Mad proteins can inhibit cellular growth and transformation and thus antagonize the function of Myc proteins. To define further the contribution of these proteins to cellular growth control, we have studied the expression of the respective genes and proteins in 3T3-L1 cells, both upon serum stimulation of quiescent cells and during adipocytic differentiation in response to insulin, dexamethasone, and isobutylmethylxanthine. We found distinct expression patterns for the mad genes. Mad4 was induced when cells exit the cell cycle and, together with mad1, during the late phase of differentiation. In contrast, mad3 expression was associated with progression through S phase and the proliferative burst of differentiating preadipocytes, overlapping in part c myc expression. DNA binding analyses revealed that the most prominent network complex both in cycling and in differentiating cells was Mnt/Max, whereas c Myc/Max complexes were detectable only during peak c-Myc expression periods. Ectopic expression of Mad1 in preadipocytes resulted in the inhibition of S phase and the proliferation associated with the proliferative burst; as a consequence, adipocytic differentiation was significantly inhibited. Our findings suggest that the precise temporal regulation of Myc/Max/Mad network proteins is critical for determining cellular behavior. PMID- 10797317 TI - Rapid lamellipodia formation in nerve growth factor-stimulated PC12 cells is dependent on Rac and PI3K activity. AB - Neuronal differentiation of PC12 cells is achieved by stimulation with nerve growth factor (NGF) but not by epidermal growth factor (EGF). However, features of differentiation such as neurite outgrowth are observable at the earliest after several hours. Using actin staining of the cells, we show here that NGF stimulation leads to lamellipodia formation within only 3 min at the periphery of the PC12 cells. EGF stimulation or microinjection of differentiation-inducing c Crk I protein does not cause lamellipodia. The actin reorganization after NGF stimulation is blocked by microinjecting dominant negative Rac protein. The lamellipodia formation is also abolished by inhibitors of phosphatidylinositol 3 kinase, wortmannin and LY 294002 in a concentration-dependent manner. Phase contrast time-lapse microscopy was used to analyze membrane dynamics in real time and to confirm the induction of lamellipodia by NGF and their inhibition by pretreatment with both wortmannin and LY 294002. The results indicate that NGF, but not EGF, leads to rapid lamellipodia formation in PC12 cells via phosphatidylinositol 3-kinase and the small GTPase Rac, thereby defining a novel role for these factors in early NGF signaling. PMID- 10797318 TI - Sensitization of tumor cells to fas killing through overexpression of heat-shock transcription factor 1. AB - Activation of the heat-shock or stress response is generally considered a cytoprotective response to heat or other proteotoxic stresses. In mammalian cells, stress-induced transcription of heat-shock genes is regulated by heat shock transcription factor 1 (HSF1). We now show that activation of the Fas death receptor transactivates HSF1 in HeLa cells, a Fas-expressing cervical carcinoma line. Whereas HSF1 is constitutively expressed in a non-DNA-binding, transcriptionally inactive state, activation of Fas leads to enhanced transcription of a heat-shock reporter gene. The effects of Fas on heat-shock gene transcription do not appear to be a consequence of cell death as they (1) precede apoptotic changes and (2) are not abrogated by YVAD-CMK, an inhibitor of Fas apoptosis that acts by blocking downstream effector proteases. Despite expressing Fas, HeLa cells are relatively insensitive to Fas-mediated killing, indicating that Fas expression alone, although necessary, is not sufficient for apoptosis. By overexpressing a constitutively activated form of HSF1, we sensitize HeLa cells to Fas-mediated killing. These findings shed new light on the interaction between two of the most evolutionarily conserved cell programs in nature, the Fas death pathway and the heat-shock response. Strategies designed to upregulate HSF1 in tumor cells, either through pharmacologic or gene-therapy approaches will hopefully provide a means with which to sensitize tumors to the killing effects of cancer therapies operating through the Fas receptor. PMID- 10797319 TI - Diel variations in respiration, excretion rates, and nutritional status of zooplankton from the Pampulha Reservoir, Belo Horizonte, MG. AB - This investigation is focused on the experimental determination of diel cycles of metabolic activity of zooplankton in a tropical reservoir. Water and zooplankton used in laboratory experiments were collected from the Pampulha reservoir. The experimental units were incubated in the light (1500 Lux) and in the dark at 25.0 +/- 1.0 degrees C during different periods of the diel cycle. At the end of each experiment, the following variables were measured: temperature, dissolved oxygen, ammonia, and orthophosphate as well as the composition, abundance and dry weight of the zooplankton. The specific respiration and excretion rates were determined considering the differences in concentration between experimental and control units. The effect of diurnal cycle on respiration rates was clearly more intense than the effect of light. The average values of respiration rates obtained in the morning hours oscillated between 0. 015 and 0.016 mgO(2)mgDW. hr(-1) (light and dark incubations). At night, these rates were higher and ranged from 0.020 to 0.035 mgO(2)mgDW. hr(-1). Increased biomass of zooplankton and longer incubation times produced lower respiration rates. The excretion rates of ammonia were higher at night, reaching a mean value of 4.2 microgN-NH(4)/mg DW. hr(-1) in illuminated units. The phosphate excretion rates were more elevated in the morning, reaching 0.58 microgP-PO(4)/mg/DW. hr(-1) illuminated vessels. The nanoplankton was able to actively absorb ammonia as well as phosphate. The highest ammonia absorption rates were measured at night, whereas the nanoplankton absorbed phosphorus only in the morning hours. The nutritional status of zooplankton also showed short-term variations. The mean phosphorus content of zooplankton biomass also varied between day and night as well as with incubation time. It ranged from 0.58-2.17%, whereas organic matter variation was more conservative, oscillating around 70-92% in all occasions. PMID- 10797320 TI - Role of nitric oxide in the systemic and pulmonary circulation of anesthetized turtles (Trachemys scripta). AB - In reptiles the influence of local vascular factors on blood flow regulation is vaguely understood. The aim of this study was to investigate the role of nitric oxide (NO) on vascular function in anesthetized Trachemys scripta. The experimental protocol consisted of serial injections of sodium nitroprusside (SNP; 25 microg. kg(-1)), L-arginine (185 mg. kg(-1)) and L-NAME (50 mg. kg(-1)). SNP induced a systemic vasodilation (0.05 to 0.02 kPa. min. kg. mL(-1), P = 0.015), with no change in pulmonary vascular resistance (0.07 versus 0.08 kPa. min. kg. mL(-1), P > 0.05). L-Arg had no effect on resistances but increased cardiac output by 17%. L-NAME increased systemic resistance (33% increase; P = 0.01) while pulmonary resistance was unchanged. These effects are consistent with in vivo and in vitro studies on the systemic vasculature of different reptilian species, suggesting that NO has an important role in maintaining systemic vascular tone. The pulmonary vasculature did not respond to NO due to either a lack of an endogenous NO tone or a relaxed state of the pulmonary vasculature. The importance of NO-based mechanisms versus other neuro-humoral modulators in the reptilian circulation remains uncertain. However, as established in prior studies, cholinergic control of the proximal pulmonary artery is the main regulator of pulmonary resistance while systemic resistance depends on a more complex suite of neural, humoral and local effectors that include NO. PMID- 10797321 TI - Immunoelectron microscope analysis of epidermal growth factor receptor (EGFR) in isolated Mytilus galloprovincialis (Lam.) digestive gland cells: evidence for ligand-induced changes in EGFR intracellular distribution. AB - In mammalian cells, the binding of epidermal growth factor (EGF) to its receptor (EGFR), a glycoprotein with intrinsic tyrosine kinase activity, leads to the pleiotropic responses to EGF. Among these, a negative feedback response by stimulation of receptor internalization and lysosomal degradation, this attenuating signal transduction. In this work, data are reported on the identification of specific EGFRs in isolated digestive gland cells from the marine mussel (Mytilus galloprovincialis Lam.) By immunoelectron microscopy. In control digestive cells, EGFR immunoreactivity was mainly associated with cytoplasmic membrane structures and, to a lesser extent, the cell membrane. The presence of EGFR-like receptors was confirmed by Western blotting of digestive gland cell extracts with two different monoclonal antibodies that recognize either intracellular or extracellular epitopes. The addition of mammalian EGF resulted in significant time and temperature-dependent changes in EGFR subcellular distribution in mussel cells. In cells exposed to EGF for 0-15 min at 4 degrees C, the distribution of EGFR was not significantly different from that of the control cells. On the other hand, at 18 degrees C, an increased labelling along the cell membrane was observed after 5-10 min after EGF addition, with a concomitant decrease in the cytoplasmic signal. Moreover, after 20 min of exposure to EGF, ligand binding apparently resulted in EGFR compartmentation within the lysosomes. These observations were confirmed by quantitative analysis of EGFR labelling at different times of EGF exposure. Similar results were obtained utilizing the two different monoclonal antibodies. The results indicate that, in mussel digestive cells, the binding of heterologous EGF to specific receptors induces a negative feedback response by stimulating the lysosomal degradation of EGFR, thus suggesting the presence of mechanisms responsible for receptor downregulation similar to those observed in mammalian cells. PMID- 10797322 TI - Anoxic performance of the american eel (Anguilla rostrata L.) heart requires extracellular glucose. AB - The importance of extracellular glucose in the maintenance of performance of the heart of the American eel (Anguilla rostrata Le Sueur (L.) Under anoxia was assessed under a variety of experimental conditions. Ventricular strips, electrically paced at 36 bpm, in N(2)-gassed medium maintained the imposed pace rate and generated approximately 25% of the initial twitch force of contraction for at least 60 min when glucose was present in the medium. But ventricular strips challenged without glucose in the medium failed to maintain the pacing rate within 5-10 min. Isolated and intact, perfused hearts maintained pressure and followed an imposed pace rate of 24 bpm for at least 2 hr, under anoxic conditions, if glucose was present in the medium. But without glucose in the medium isolated hearts failed within 30 min. Endogenous glycogen stores were utilized in hearts perfused with medium containing NaCN to impair oxidative phosphorylation. The presence of glucose in the medium did not protect against glycogen mobilization. The data indicate that exogenous glucose is necessary to maintain performance under anoxia at high workloads and physiological Ca(2+) levels. Finally, ventricular strips treated with NaCN and forced to contract at 24 bpm lost 70% of initial twitch force. Increasing extracellular Ca(2+) concentration stepwise from 1.5 to 9.5 mM restored twitch force to approximately 50% of the initial level and this response was not dependent on exogenous glucose. However, glucose was required to maintain resting tension even under normoxic conditions in the face of a Ca(2+) challenge. PMID- 10797323 TI - Maintenance of resting tension in the american eel (Anguilla rostrata L.) heart is dependent upon exogenous fuel and the sarcoplasmic reticulum. AB - The relationship between extracellular glucose and management of cell Ca(2+) in the heart of the American eel (Anguilla rostrata) was indirectly assessed by monitoring the performance of isolated ventricular strips at 20 degrees C. Twitch force increased in ventricular strips under specific conditions of 30 bpm pacing and an extracellular Ca(2+) challenge from 1.5 to 9.5 mM. The response was independent of any exogenous metabolic fuel in the medium. Resting tension was maintained when glucose was available, but in the absence of a metabolic fuel, resting tension increased in response to the increase in extracellular Ca(2+) level. When ventricular strips were treated with iodoacetate to inhibit glycolysis, a Ca(2+) challenge resulted in a decrease in twitch force in association with an approximately equivalent increase in resting tension even in the presence of exogenous glucose. However, when pyruvate (5 mM) was substituted as a metabolic fuel, twitch force increased as a function of extracellular Ca(2+), and resting tension was maintained in the presence of iodoacetate. Therefore, there is a need for an extracellular fuel but not a specific metabolic requirement for glucose to maintain the performance characteristics, which are presumably related to the management of intracellular Ca(2+) levels. Ventricular strips were treated with ryanodine to inhibit Ca(2+) release and uptake by the sarcoplasmic reticulum (SR). Ryanodine treatment impaired postrest potentiation at high extracellular Ca(2+) levels. In the presence of ryanodine, the protective effect of glucose on the increase in resting tension in the face of an extracellular Ca(2+) challenge was eliminated. Considered together, the results reveal that the heart of the American eel has a requirement for an extracellular fuel to manage intracellular Ca(2+) at high Ca(2+) loads, and that the SR plays a role in the beat-to-beat regulation of Ca(2+) at a frequency of 30 bpm, high Ca(2+) load, and 20 degrees C. PMID- 10797324 TI - Evidence for leptin expression in fishes. AB - Tissues from bony fish were screened with anti-mouse leptin antibodies to detect the presence of the fat-regulating hormone in fishes. Low molecular-weight (16 kDa) immunoreactive bands were detected in blood, brain, heart and liver of green sunfish (Lepomis cyanellus), bluegill sunfish (Lepomis macrochirus), largemouth bass (Micropterus salmoides), white crappie (Pomonix annularis), channel catfish (Ictalurus punctatus), and rainbow trout (Oncorhynchus mykiss). To further verify that we had identified leptin, the response of fish "leptin" was measured in fed and fasted green sunfish. Fed sunfish had approximately threefold higher concentration of leptin in blood than did fasted sunfish (fed vs. fasted; 0.599 +/- 0.03 microg/microl vs. 0.196 +/- 0.04 microg/microl; P > F = 0.0001), which is consistent with mammalian models of leptin function. Brain leptin concentration is also positively correlated with percent body fat in white crappie and bluegill. Based upon electrophoretic mobility, immunoreactivity, response to fasting, and correlation with adiposity, we believe we have the first evidence for leptin expression in an ectotherm. PMID- 10797325 TI - Ecdysteroid hormones and metabolites of the stone crab, Menippe mercenaria. AB - The Y-organs of the xanthid crab Menippe mercenaria secrete the ecdysteroids, 3 dehydroecdysone (3DE) and lesser amounts of 3-dehydro (or 2-dehydro)-25 deoxyecdysone (3D25dE) in vitro. These ecdysteroids were identified by elution time comparisons with authentic standards, mass spectrography, and, for 3D25dE, infrared spectrometry. Tissues were incubated 18 hr with [(3)H]3DE. Activities representing 3beta-reductase and 20-hydroxylase generally were present, evidenced by finding in the tissue/medium extract labeled ecdysone (E) and 20 hydroxyecdysone (20E). Labeled 3-dehydro-20-hydroxyecdysone (3D20E) also appeared to be present. Tissue blanks and hemolymph were devoid of activity. Muscle was low, hypodermis was intermediate, and hindgut and gonads were high in activity of the enzymes. Consistent with the presence of these enzymes in peripheral tissues, ecdysteroid products identified in the hemolymph were 20E, 3D20E, and 25-deoxy-20 hydroxyecdysone (25d20E; ponasterone A). Structures of 20E and 3D20E were confirmed by co-elution with authentic standards in high-performance liquid chromatography (HPLC), co-elution of derivatives in gas chromatography, and mass spectroscopy. Ponasterone A (identified by HPLC co-elution with the standard), like 20E is present in the hemolymph in prominent amounts. These data indicate that Menippe, among crustaceans thus far studied, secretes a unique combination of ecdysteroid hormones, namely, a 3- (or 2-) oxo compound and a 25-deoxy compound. This represents a different kind of branch point from 5beta-diketol in ecdysteroid biosynthesis, in which the intermediate, 5beta-ketodiol is bypassed. A result is the joint appearance in the circulation of the hormones, 20E and ponasterone A, which in other species are singly prominent. PMID- 10797327 TI - Female gonad of moles, genus Talpa (Insectivora, mammalia): ovary or ovotestis? AB - The female gonads of moles (genus Talpa) are composed of a cortex, functioning as an ovary, and a medulla, which is structurally similar to that of the testis. In the female reproductive apparatus there are masculine glandular annexes, such as a bilobate prostate, two Cowper glands, and a penis-like clitoris. All these features have recently led to the hypothesis of the presence of hermaphroditism due to sex-reversal in Talpa. The purpose of this study is to understand the functional significance of the structural organization of the female gonads in order to verify this hypothesis. Histological, histochemical and ultrastructural analyses have been carried out on several gonads of both sexes of two species: T. europaea and T. romana, including three fetuses. In both species, the cortical region of the female gonad shows a regular oogenetic activity. While the medulla is composed of interstitial cells that are partly organized in cord-like structures, no spermatogenetic activity has been ever observed inside of them. A histochemical analysis shows that in both sexes the interstitial cells secrete steroids, presumably estrogens as well as androgens. The presence of androgens in the female gonads would therefore explain the persistence of male glandular annexes in the female reproductive apparatus and both the sexual and behavioral monomorphism typical of the genus Talpa. Nonetheless, the female gonad of moles is a real ovary and a well-defined gonochorism exists. Therefore, there is no reason to assert the presence of hermaphroditism due to sex reversal. PMID- 10797326 TI - Ontogeny of B cells expressing IgM in embryonic and larval tissues of the American grass frog, Rana pipiens. AB - Affinity-purified, fluorochrome-tagged F(ab')(2) antibody fragments specific for heavy (mu) chains of Rana pipiens IgM were prepared from hyperimmune rabbit sera. By using two-color immunofluorescent procedures we observed that (1) the first cells expressing IgM, termed pre-B cells, lack detectable quantities of membrane or surface IgM but contain detectable quantities of cytoplasmic IgM (smu( )/cmu(+)), (2) sIgM(+) B cells were the second type of IgM containing cell to appear in development, and (3) plasma cells, which contain copious quantities of cIgM, were the final phenotype to appear in the development of B cells expressing IgM. These cells were first observed in the pronephros of the developing urogenital system. Shortly after their appearance in the pronephros, cells in B lineages were observed in the liver. These observations (1) are consistent with recent studies of B lymphopoiesis in the aorta-gonad-mesonephros (AGM) region in endothermic vertebrates, including mice, (2) suggest that there are fundamental ontogenetic and phylogenetic similarities between cells and tissues of developing vertebrate immune systems, and (3) evoke questions concerning the possible function(s) of lymphocytes in developing anurans up to metamorphosis and beyond. PMID- 10797328 TI - Polymerization of a vitelline envelope (VE)-like structure produced by using fractionated VE extracts from carp eggs. AB - Fractionation of vitelline envelope (VE) extracts from carp eggs made possible the efficient polymerization of a VE-like structure. The structure corresponded to the fourth layer of the VE or fertilization envelope (FE), and its organization was achieved by reassembly in vitro after solubilization of the sheets composed of filamentous substances or network-like aggregates which were induced by a cortical alveolus sialoglycoprotein or thrombin. The sialoglycoprotein was a serine proteinase and immunolocalized only in the structure at the periphery of cortical alveoli, not in the VE and yolk granules. Ultrastructural features of the VE-like structure suggested that reassembly in vitro occurred via several intermediates in the process of polymerization. A polyclonal antibody produced against one of the assembled VE components, a 64 kDa protein, more intensely immunostained the outer periphery of the VEs than other areas, and immunoelectron microscopy showed that immunogold particles specifically labeled reassembled VE-like structures and major skeletons of the networks or network-like sheets. The protein with a molecular weight of 64 kDa was found to be a DNase. Thus, these results suggest a new approach to investigating not only the FE assembly process in vitro but also the organizing relationship between the major skeleton of the VE or FE and other additional constituents. PMID- 10797329 TI - Evidence for direct membrane retrieval following cortical granule exocytosis in Xenopus oocytes and eggs. AB - Rapid exocytosis is typically followed by rapid resorption of exocytosed membrane; however, whether membrane retrieval occurs via indirect endocytosis of numerous small vesicles or direct resealing of the original, larger exocytotic vesicles is controversial. Here we show that cortical granule (CG) exocytosis in Xenopus oocytes and eggs is followed by rapid formation of endosomes as large as the CGs. Large endosomes are translucent, and their formation has the same developmental and pharmacological profile as CG exocytosis. Time course analyses show that large endosomes are not derived from small endosomes. Large endosome formation is triggered by stimuli that do not trigger increases in intracellular free calcium and is insensitive to perturbation of microtubules by treatment with nocodazole. Perturbation of the f-actin cytoskeleton with latrunculin, however, sharply reduces large endosome formation. We conclude that CG membrane is directly retrieved in Xenopus oocytes and eggs and suggest that this retrieval is not directly dependent on an increase in intracellular-free calcium, but is dependent on the actin cytoskeleton. PMID- 10797330 TI - The national cancer data base: what does it mean to the community surgeon? PMID- 10797331 TI - Quality of life in survivors with a Van Ness-Borggreve rotationplasty after bone tumour resection. AB - BACKGROUND AND OBJECTIVES: In large malignancies of the distal femur, limb salvage may not be feasible. Amputation is often the treatment of choice. A Van Ness-Borggreve rotationplasty is an alternative when the sciatic nerve is free of tumour. The appearance of the rotated lower leg is poor, which justifies a general concern about the psychosocial functioning of these patients. The aim of the study was to assess the medium- and long-term effects on quality of life (QOL) after rotationplasty. METHODS: A self-report questionnaire included the SF 36, the Social Support List, and selected items from the European Organisation for Research and Treatment of Cancer (EORTC) questionnaires as well as study specific questions. The questionnaire was mailed to 34 patients at once. All patients were older than 16 years and at least 1 year postsurgery (mean 6.3 years). The response rate was 96%. RESULTS: Patients' physical functioning was poorer than that of healthy peers but better in comparison to chronically ill patients. Levels of psychosocial functioning, general QOL, and social support were highly comparable with those of healthy peers. One-third to one-half of the patients reported negative effects of the surgery on initiating social and/or intimate contacts, body image, and sexuality. With respect to physical functioning, two-thirds of patients engaged actively in sports. Patients reported wearing the prosthesis continuously and were, in general, satisfied with its fit. CONCLUSIONS: Given the relatively high levels of QOL and psychosocial functioning of these patients, these results indicate that rotationplasty is a good alternative in the treatment of patients with a large malignancy of the distal femur. PMID- 10797332 TI - Function and complications after ablative and limb-salvage therapy in lower extremity sarcoma of bone. AB - BACKGROUND AND OBJECTIVES: The functional results and the complications after several limb-saving and ablative treatments because of lower extremity bone sarcoma were evaluated. METHODS: Seventy-seven surviving patients were evaluated according to the MSTS (American Musculoskeletal Tumor Society) functional rating system. Fifty-two patients had limb-saving and 25 had ablative therapy. Median follow-up was 97 months in the limb-saving group and 112 months in the ablative group. RESULTS: Functional results in the limb-saving group were significantly better than in the ablative group (P = 0.0001). Functional results in patients with tumors about the knee joint were significantly better (P = 0.0064) after limb-saving surgery (i.e., endoprosthesis, knee arthrodesis, or rotationplasty) compared to functional results after ablative surgery (i.e., hip or knee disarticulation or above-the-knee amputation). Complications were 3 times more common after limb-salvage procedures and 4 times more common after endoprosthetic reconstructions compared to after ablative procedures. Complications after limb saving therapy were fewest in tumors about the knee joint. In 3/28 patients, the endoprosthetic reconstruction had to be converted to an amputation. CONCLUSIONS: Functional results were significantly better after limb-saving compared to after ablative therapy. Complications, however, were more common after limb-saving therapy. PMID- 10797333 TI - Predictors of functional outcomes following limb salvage surgery for lower extremity soft tissue sarcoma. AB - BACKGROUND AND OBJECTIVES: Patient function has been conceptualized by clinical measures such as joint motion, muscle strength, disability, and general health status. The purpose of the current study was to evaluate tumor and treatment variables predictive of these conceptually different posttreatment functional outcomes in patients treated with limb preservation surgery for lower-extremity soft tissue sarcoma. METHODS: One hundred seventy-two patients with minimum 1 year follow-up were evaluated using the following outcomes: impairment, measured by the 1987 and 1993 versions of the Musculoskeletal Tumor Society Rating Scale (MSTS); disability, measured by the Toronto Extremity Salvage Score (TESS); and general health status, using the Short Form-36 (SF-36). Tumor and treatment related variables (age, gender, presenting disease status, anatomic site, tumor size, grade, depth, prior excision, irradiation, bone resection, motor nerve sacrifice, and complications) were extracted from the STS database. RESULTS: Large tumor size, bone resection, motor nerve resection, and complications were predictive of lower MSTS 1987 and 1993 scores. Patients with large, high-grade tumors who required motor nerve resection were more disabled, as reflected by lower TESS scores. Only age and prior surgery were adverse predictors of SF-36 score. CONCLUSIONS: These results demonstrate that different factors are predictive of different patient outcomes, specifically, impairment, disability, and general health status. It is important to define function when counseling patients regarding their potential recovery based on tumor and treatment-related variables. J. Surg. Oncol. 2000;73:206-211. PMID- 10797334 TI - Factors influencing survival after resection for ductal adenocarcinoma of the head of the pancreas. AB - BACKGROUND AND OBJECTIVES: Recent reports have demonstrated improvement in the 5 year actuarial survival for patients with resected ductal adenocarcinoma. The purpose of this study is to determine the factors favoring long-term survival after pancreaticoduodenectomy. METHODS: Between 1974 and 1995, 75 patients with pancreatic head carcinoma underwent pancreaticoduodenectomy in our department. RESULTS: Overall postoperative mortality rate was 5. 3% and morbidity was 24%. Median survival following resection was 17 months. Estimated 1-, 2-, and 5-year survival rates were 68%, 46.7%, and 18.7%, respectively. Five-year survival was greater for node-negative than for node-positive patients (41.7% vs. 7.8%, P < 0. 001) and for smaller (<3 cm) than for larger tumors (33.3% vs. 8.8%, P < 0.006). The 5-year survival in patients with negative margins (n = 60) was 23.3%, whereas no patient with positive margins (n = 15) survived at 13 months (P < 0.001). Multivariate analysis, performed by the Cox proportional hazards model, indicated that margin status, lymph node metastasis, tumor size, and poor histological differentiation were independent predictors of poor survival. CONCLUSIONS: Five year survival for patients undergoing pancreaticoduodenectomy for ductal adenocarcinoma of the pancreas was 18.7%. Survival was greater in the group of patients with negative lymph nodes, tumor size <3 cm, and negative margin status. PMID- 10797335 TI - Telomerase activity in colorectal cancer and its relationship to bcl-2 expression. AB - BACKGROUND AND OBJECTIVES: Telomerase is thought to be responsible for cell immortality, and bcl-2 has been demonstrated to regulate apoptosis. Recent studies have shown a wide occurrence of telomerase activation and bcl-2 deregulation in human carcinoma cells. METHODS: We examined telomerase activity in tissues from 50 patients with colorectal carcinoma with a telomeric repeat amplification protocol assay. We also investigated the relationship between telomerase activity and expression of bcl-2 in 37 colorectal carcinoma specimens. RESULTS: We detected telomerase activity in 33 (66%) of 50 colorectal carcinomas, whereas no activity was detected in the adjacent noncancerous mucosa of 13 tumor specimens. There was no correlation between pathological stage and telomerase activity. Telomerase activity in the bcl-2-expressing cases was higher than that in the bcl-2-non-expressing cases. CONCLUSIONS: Expression of bcl-2 may be related to telomerase activity in colorectal carcinomas. PMID- 10797337 TI - Benign chondroid syringoma: report of a case clinically mimicking a malignant neoplasm. AB - Chondroid syringoma, or mixed tumor of skin, is an uncommon sweat gland tumor most often seen in the head-and-neck region of patients in the sixth or seventh decade. Tumors usually present as asymptomatic, slowly growing masses. Histologically, there are both epithelial and stromal components. The treatment of choice is local excision. Rare malignant examples have been reported, commonly involving the extremities. We present a case of cutaneous chondroid syringoma arising in the thigh of a 28-year-old female. The tumor grew over a 4-year period, increasing rapidly in size over the last few months with fixation and pigmentation of the overlying skin clinically mimicking a malignant neoplasm. Such cutaneous appendage tumors are uncommon, and surgeons may be unfamiliar with them. PMID- 10797336 TI - Intraoperative topical tetracycline sclerotherapy following mastectomy: a prospective, randomized trial. AB - BACKGROUND AND OBJECTIVES: Postoperative wound seromas are a frequent and troublesome occurrence after mastectomy. Recent reports have suggested the efficacy of topical sclerosants at reducing their formation. METHODS: A prospective, randomized, double-blinded trial was performed to examine the effect of intraoperatively administered topical tetracycline on the occurrence of postoperative mastectomy seromas. Thirty-two women were randomized to the control arm (normal saline) and 30 women to the tetracycline arm. In the treatment group, 100 ml (2 g) of tetracycline solution was administered topically to the chest wall and skin flaps prior to skin closure. The control group received an equal volume of normal saline. Patients were monitored for the development of postoperative wound seroma. RESULTS: There were no significant differences between groups regarding total volume of closed suction drainage, numbers of patients leaving hospital with drains in place, or duration of catheter drainage. Seroma formation 2 weeks postoperatively was greater in the tetracycline group than the control group (53% vs. 22%, P = 0.01). There were no differences between groups regarding the degree of postoperative pain, wound infection, or seroma formation 1 month postoperatively. CONCLUSIONS: Topical tetracycline is not effective at preventing post-mastectomy wound seromas. PMID- 10797338 TI - Pregnancy-associated melanoma occurring in two generations. AB - We report of a case of malignant melanoma occurring during pregnancy in a woman whose mother had a melanoma excised during pregnancy. There was no other family history of melanoma. To our knowledge, this has not been previously reported. A review of the recent literature suggests that pregnant women with melanoma do not have a worse prognosis when compared to matched controls, but may present with worse prognostic features. PMID- 10797339 TI - Radiofrequency ablation of metastatic liver tumor. PMID- 10797340 TI - Prevention of postoperative bile leakage after major hepatic resections in malignancies. PMID- 10797341 TI - Surgical oncology of the pelvis: ostomy planning and management. AB - Advanced pelvic surgery may result in the formation of a colostomy, ileostomy, urostomy, or multiples of these stomas. Stomas may be created because of a planned therapeutic attack to eradicate cancer or manage complications arising from treatments or they may be constructed to palliate severe symptoms or life threatening problems. In some instances, fistulae between various organs and the skin may occur. Special skills are required to meet the needs of patients with ostomies or fistulae. The often complex care presents critical challenges to nurses, particularly in today's cost-conscious health care environment where prompt hospital discharges, brief clinic appointments, and valuable but limited home care visits are the norm. The skills of the enterostomal therapy nurse are valuable in all phases of care of these patients, from preoperative to outpatient care. PMID- 10797342 TI - Groin dissection. AB - This review provides a brief history of groin dissection, including studies on anatomical considerations and technique. A groin dissection for complete ablation of the node-bearing areolar tissue in the inguinal and iliac regions and with negligible morbidity requires careful attention to the pathophysiology of cancer in lymphatics, pre- and postoperative care, and surgical technique, including coordinated use and general, plastic, vascular, and orthopedic surgical principles. PMID- 10797343 TI - Vascular problems of the pelvis. AB - Vascular tumors of the pelvis are a rare, diverse group of neoplasms. These benign or malignant tumors can arise from the endothelium, smooth muscle cells, or pericytes of the arterial venous or lymphatic walls. They are rarely diagnosed by physical examination but more commonly seen with imaging studies such as computed tomography, magnetic resonance imaging, or angiography. Benign and malignant tumors can be differentiated pathologically by the two major anatomic characteristics of vascular channel formation and the regularity of endothelial cell proliferation. This review will focus on the clinical presentation, diagnosis, treatment, outcomes, and surgical approaches to benign and malignant vascular tumors of the pelvis. PMID- 10797344 TI - Survival of patients with untreated breast cancer. AB - Knowing the clinical prognosis of untreated breast cancer is useful in dealing with patients with neglected disease or in environments with poorly developed healthcare systems. This study analyzes historical survival data in two sets of untreated patients: (1) 250 patients followed until death (up to 12 years) for which autopsy results are available and (2) an amalgam of 1,022 patients from several papers. Data from nine published papers underwent actuarial analysis. Median survival time of the 250 patients followed to death was 2.7 years. Actuarial 5- and 10-year survival rates for these patients with untreated breast cancer was 18.4% and 3.6%, respectively. For the amalgamated 1,022 patients, median survival time was 2.3 years. Actuarial 5- and (partially fitted) 10-year survival rates for these patients with untreated breast cancer was 19.8% and 3.7%, respectively. Historical data of untreated breast cancer patients reveal a potential for long survival in some cases. The spectrum of clinical aggressiveness of breast cancer varies between virulence and chronic disease. PMID- 10797345 TI - Biological considerations with pelvic neoplasms. PMID- 10797346 TI - Reply. PMID- 10797347 TI - Tamara Dembo's European years: working with Lewin and Buytendijk. AB - In this paper early work of the American rehabilitation psychologist Tamara Dembo (1902-1993) is brought to light. She was highly influenced by the concepts of Kurt Lewin's topological psychology, and she used the framework of topological psychology to analyze her investigations on animal behavior carried out with the Dutch zoopsychologist Frederik J. J. Buytendijk. These investigations have so far been ignored and are being described for the first time making use of archival materials. PMID- 10797348 TI - The concept of social class: the contribution of Everett Hughes. AB - In French Canada in Transition (1943) and a set of related essays written between 1933 and 1941, Everett Hughes, a key figure in the "Second Chicago School" of sociology developed a novel and noteworthy conceptualization of social class. This contribution, which was not recognized outside of French-language sociology in Quebec, was an integral element of Hughes's "interpretive institutional ecology" theoretical frame of reference. It combined elements of the classical ecological theory of class (human ecology, functionalism, Simmel), aspects of a Weber-inspired analysis of class, status, and political power, and elements of a proto-dependency analysis of Quebec's industrialization in the 1930s. PMID- 10797349 TI - Adaptive will: the evolution of attention deficit disorder. AB - The increasing prevalence of attention-deficit disorder among American school children was a source of significant controversy in the 1990s. This paper looks at the social and historical contexts in which ADD evolved in order to understand its emergence as a coherent and widespread entity. Changes in expert models of child behavior interacted with the formation of new identities around disability to shape a milieu in which the disorder could thrive. The pattern of affect control, of what must and what must not be restrained, regulated, and transformed, is certainly not the same in this stage as in the preceding one of court aristocracy. In keeping with its different interdependencies, bourgeois society applies stronger restrictions to certain impulses, while in the case of others aristocratic restrictions are simply continued and transformed to suit the changed situation (Elias, 1994, p. 125). PMID- 10797352 TI - Novel multiagent chemotherapy for bone marrow relapse of pediatric acute lymphoblastic leukemia. AB - BACKGROUND: Despite improvements in the treatment of pediatric acute lymphoblastic leukemia, approximately one in five patients will develop recurrent disease. The majority of these patients do not survive. This limited institution study sought to improve event-free survival (EFS) by intensification of chemotherapy. PROCEDURE: Twenty-one patients with either an isolated marrow (n = 16) or a combined marrow and central nervous system relapse (n = 5) received treatment according to Children's Hospital of Philadelphia protocol CHP-540. Six patients had an initial remission of <36 months, and five patients had relapsed within 1 year of completion of phase III therapy. Induction and reinduction therapy consisted of idarubicin, vincristine, dexamethasone, asparaginase, and triple intrathecal chemotherapy. Consolidation and reconsolidation therapy employed high-dose cytarabine, etoposide, and asparaginase given in a sequential manner. Maintenance therapy included courses of high- or low-dose cytarabine followed by sequential etoposide and asparaginase pulse, moderate-dose methotrexate with delayed leukovorin rescue, and vincristine/dexamethasone pulses. Therapy continued for 2 years from the start of interim maintenance in the 16 patients who did not receive a bone marrow transplant (BMT). Two patients underwent an HLA-identical sibling BMT specified by protocol. Four received a nonprotocol-prescribed alternative donor BMT. RESULTS: The complete remission induction rate was 95%. With a median follow-up from date of relapse of 49 months in survivors, the actuarial EFS based on intent to treat is 75%. There were three toxic deaths in patients in CR and two deaths from relapse. CONCLUSIONS: This regimen is toxic but effective and deserves study in a larger setting. PMID- 10797353 TI - Granulocyte-macrophage colony-stimulating factor support in therapy of high-risk acute lymphoblastic leukemia in children. AB - BACKGROUND: Our purpose was to increase the dose intensity of chemotherapy and reduce the days with neutropenic fever in childhood high-risk (HR) acute lymphoblastic leukemia (ALL) by systematic use of granulocyte-macrophage colony stimulating factor (GM-CSF). PROCEDURE: All children with HR-ALL in Finland during 1990-1996 were included. Two open-label study groups were formed: 1) 34 children diagnosed between January, 1992, and December, 1996, received seven or nine courses (depending on cranial RT or no cranial RT) of GM-CSF at 5 microg/kg s.c. daily until an absolute neutrophil count (ANC) of 1,000 x 10(6)/liter at scheduled places in the protocol and 2) 80 control children, those diagnosed between January, 1990, and December, 1991, plus all with significant coexpression of myeloid markers, did not receive GM-CSF. RESULTS: Dose intensity increased in patients who received regular GM-CSF support. The intensive phase of therapy, including induction, consolidation courses, and delayed intensification, was 33 days shorter (P < 0.001) in children with seven courses and 26 days shorter (P < 0.01) in those with nine courses of GM-CSF compared to controls. The number of infections during the whole ALL therapy was reduced by use of GM-CSF in children aged >5 years (P < 0.001), but not in those aged <5 years. The mean total duration of intravenous antibiotics per child was 39 days in the GM-CSF group and 48 days in the control group (P < 0. 001). Systematic use of GM-CSF was cost effective. CONCLUSIONS: Systematic use of GM-CSF improved dose intensity by shortening the intensive treatment period by about 4 weeks. Use of GM-CSF reduced the days for inpatient antibiotics by about 1 week per child, which translates into reduced costs. PMID- 10797354 TI - Busulfan, melphalan, and thiotepa with or without total marrow irradiation with hematopoietic stem cell rescue for poor-risk Ewing-Sarcoma-Family tumors. AB - BACKGROUND: Survival following metastatic or recurrent Ewing sarcoma family tumors (ESFT) remains <25%. Myeloablative therapy with hematopoietic stem cell transplantation (HSCT) may improve survival for poor-risk ESFT. We describe the toxicity and efficacy of a myeloablative chemotherapy regimen, followed by a second myeloablative radiotherapy regimen as consolidation treatment for poor risk ESFT. PROCEDURE: Sixteen patients with poor-risk ESFT were treated with myeloablative therapy followed by HSCT. All patients received busulfan, melphalan, and thiotepa (BuMelTT) as chemotherapy conditioning. Nine patients received total marrow irradiation (TMI) as a second myeloablative therapy, also followed by HSCT. Seven patients were excluded from TMI because of inadequate peripheral blood stem cell harvest, extensive prior radiation therapy, early disease progression, orpatient refusal. The disease status prior to my eloablative therapy was first complete response (CR1) in three patients, CR2 in nine, second partial response (PR2) in one, CR3 in one, and progressive disease (PD) in two. RESULTS: One patient died of regimen-related toxicity, one from late pulmonary toxicity, and one following allogeneic transplantation for myelodysplasia. Eight developed recurrent disease (median time to progression 6.8 months). Six survive without relapse from 27 to 66 months following BuMelTT (median follow-up 42 months), all of whom received both BuMelTT and TMI patients (3-year event-free survival 36%). CONCLUSIONS: Dual myeloablative therapy with BuMelTT and TMI was a feasible and promising treatment approach for patients with poor-risk ESFT. Inability to collect sufficient PBSC and extensive previous radiation therapy limit the ability to deliver TMI as a second HSCT conditioning regimen. PMID- 10797355 TI - Desmoplastic small round cell tumour in children and adolescents. AB - BACKGROUND: Desmoplastic small round cell tumour (DSRCT) is a rare highly aggressive neoplasm, and clinical studies are scarce. PROCEDURE: We report six cases of children and adolescents (median age 14 years, range 6.9-17.5) with DSRCT (5 abdominal, 1 paratesticular) registered by the Italian Cooperative Group (ICG) for soft tissue sarcoma over a 9-year period. Patients received a multidisciplinary treatment, including aggressive initial or delayed surgery and radiotherapy. Chemotherapy regimen was based on the use of ifosfamide, vincristine, dactinomycin, and a few doses of antharacyclines (doxorubicin or epirubicin). RESULTS: Complete surgical resection was possible only for the paratesticular tumour. Among the patients with abdominal lesions, macroscopically radical excision was possible in only one case. All patients received multidrug chemotherapy, and tumour reduction was obtained in the 4 evaluable patients. No relapses were evident in the irradiated fields in the 4 patients who received radiotherapy. Two patients remained progression-free 22 and 63 months after diagnosis, one is in third complete remission, whereas three died 10-25 months after diagnosis. CONCLUSIONS: DSRCT is a chemosensitive tumour, but survival rates remain disappointing despite aggressive multimodality therapy. Our results support surgical tumour removal and radiotherapy to achieve local control. Our experience and a review of the literature suggest that patients with localised abdominal tumours or a paratesticular primary may have a better prognosis. PMID- 10797356 TI - Early deaths in childhood cancer. AB - BACKGROUND: Deaths prior to or shortly after the diagnosis of childhood cancer may reflect inadequacies in detection and appropriate referral for care. This study was performed to determine the extent of and factors associated with early death in childhood cancer. PROCEDURE: Patients with of primary cancer, aged <20 years at diagnosis, were identified from the SEER data (n = 23,470) from 1973 to 1995. Early deaths were defined as cases identified by 1) death certificate, 2) autopsy report, or 3) death within 1 month of initial diagnoses (n = 481). Cause of death was determined by ICD-8 and -9 codes. Age at diagnosis, year of diagnosis, morphology, site of disease, race, and gender were evaluated for association with early death. RESULTS: Age <1 year at diagnosis (6.2% early deaths), being diagnosed earlier in the observation period, and a diagnosis of a brain tumor, neuroblastoma, leukemia, or liver tumor were associated with increased early death. Gender and race were not associated with early death. Among the cases for whom the malignant diagnosis was made at the time of death (n = 119), the cause of death was nonmalignant for 36. For 22 of these cases the malignancy was an incidental finding and appeared not to contribute directly to the cause of death. Among these patients, 11 had neuroblastoma, 9 being <1 year of age. CONCLUSIONS: A decrease in the proportion of early deaths associated with childhood cancer has occurred during the past 2 decades. This decrease may reflect earlier diagnosis or improved imaging capabilities, surgical techniques, medical therapy, and supportive care. Awareness among pediatricians, general practitioners, and emergency physicians is warranted, with a focus on high-risk groups for early detection among childhood cancer patients. PMID- 10797357 TI - Basic methods and the developing structure of a late effects surveillance system (LESS) in the long-term follow-up of pediatric cancer patients in Germany. For the German Late Effects Study Group in the German Society Pediatric Oncology and Hematology (GPOH). PMID- 10797358 TI - Weekend courses for families who have lost a child with cancer. PMID- 10797359 TI - Successful treatment of Fusarium (spp.) infection in a child with acute lymphoblastic leukemia. PMID- 10797360 TI - Epstein-Barr virus-associated lymphoproliferative disorder in a child undergoing therapy for localized rhabdomyosarcoma. PMID- 10797361 TI - Diagnosis and successful treatment of childhood primary leptomeningeal lymphoma. PMID- 10797362 TI - Minimally differentiated acute myeloid leukemia (AML-M0) in children: a single center experience. PMID- 10797363 TI - Serum type III procollagen in children with hepatoblastoma. PMID- 10797364 TI - Acute lymphoblastic leukemia and urticaria pigmentosa in an infant. PMID- 10797365 TI - Intracranial mesenchymal chondrosarcoma in an infant. PMID- 10797366 TI - Relapsing nasal glioma in a three-week-old infant. PMID- 10797367 TI - Streptokinase infusion for asparaginase-induced arterial thrombosis. PMID- 10797368 TI - Central nervous system leukemia recurring over more than 16 years. PMID- 10797369 TI - Hodgkin disease following treatment of childhood rhabdomyosarcoma. PMID- 10797370 TI - Astrocytoma following testicular cancer in a young adult. PMID- 10797371 TI - Genetically and epidemiologically related "non-syncytium-inducing" isolates of HIV-1 display heterogeneous growth patterns in macrophages. AB - The objective of this study was to identify phenotypic parameters that could distinguish among seemingly homogeneous non-syncytium-inducing (NSI) viruses and that might provide a surrogate marker for clinical progression in pediatric human immunodeficiency virus type 1 (HIV-1) infection. We undertook a pilot analysis of 15 independent HIV-1 isolates collected prospectively from two mothers and their four children who displayed a spectrum of disease stages ranging from CDC categories A1 to C3. Viruses were evaluated for their ability to replicate in primary cells (including monocyte-derived macrophages [MDM]) and cell lines, for their co-receptor preference and for genetic features of the V3 hypervariable domain of env. Virtually all isolates displayed NSI phenotypes that were restricted in their capacity to replicate in cell lines and displayed V3 loops with uniformly low net positive charges. NSI viruses from two symptomatic children and one mother were macrophage-tropic, whereas NSI isolates from two asymptomatic children were unable to replicate in MDM and were designated primary lymphotropic viruses. Only one isolate was syncytium-inducing (SI), replicated in a variety of cell lines and in MDM, used multiple co-receptors, and was dual tropic, rather than a mixture of T-cell tropic and M-tropic viruses, as assessed by genetic analysis. Phenotypic heterogeneity among NSI viruses is revealed in the ability of isolates to replicate in MDM. This characteristic is related to disease stage and provides a potentially new in vitro criterion to distinguish among NSI isolates that is unlinked to other surrogate markers. PMID- 10797372 TI - Prevalence of drug resistant mutants and virological response to combination therapy in patients with primary HIV-1 infection. AB - Baseline genotype resistance analysis was carried out in 48 adults with primary HIV-1 infection between 1995 and 1998 before starting early combination therapy. Seventeen percent (8/48) of the isolates displayed key mutations conferring resistance to reverse transcriptase (RT) inhibitors such as amino acid substitutions 215Y/F (5/48,10%), 70R (3/48, 6%), 184V (2%). Two percent (1/48) had a major mutation associated with resistance to protease inhibitors (D30N). Other mutations at positions 10, 15, 20, 33, 36, 46, 63, 71, 77, 82, 93 of the protease gene were frequent (73%). Among the 46 patients who were given antiretroviral combination therapy and who responded durably to treatment after 6 and 12 months, there was no significant difference between those harboring RT mutant strains (Group I) and those with wild-type isolates (Group II). No significant difference was found at months 6 and 12 between the two groups in terms of CD4+ cell counts. These findings suggest that the presence of drug resistant strains at the time of primary HIV-1 infection does not necessarily predict drug failure. Other factors, such as adherence to treatment, tolerance and pharmacokinetics parameters are probably major determinants of virological response in patients with early therapeutic intervention. PMID- 10797373 TI - Problems in the interpretation of HIV-1 viral load assays using commercial reagents. AB - During routine monitoring of human immunodeficiency virus (HIV) viral load, two problems arose. First, a number of patients, the majority being African, were found to have low viral loads by the Chiron branched-chain DNA assay in conjunction with low CD4(+) cell numbers. In order to determine whether this was due to failure of the branched-chain DNA assay to detect non-B subtypes of HIV, selected samples were subtyped and HIV RNA quantified by branched-chain DNA, NASBA, and the Roche Monitor RT-PCR assay. Twenty-eight (97%) of 29 Africans were infected with a non-B subtype of HIV and 15 (93.7%) of 16 non-Africans with subtype B. Twenty-three samples had a low viral load by branched-chain DNA, which was confirmed by the NASBA and RT-PCR assays. All three assays detected B and non B subtypes with similar efficiency; NASBA failed to detect HIV RNA in a small number of non-B samples. Discrepancies between viral load and CD4(+) cell numbers did not appear therefore to be related to subtype. Second, while quantification of HIV RNA was being conducted using version 2 of the branched-chain DNA assay (lower detection limit 500 HIV RNA copies/ml) the manufacturers had developed a more sensitive assay and a comparative evaluation was therefore conducted. In approximately 30% of samples the viral load was up to 10 times higher with the more sensitive assay. These experiences emphasise the importance of close collaboration between the clinic and the laboratory. PMID- 10797374 TI - Lack of protection against vertical transmission of HIV-1 by interferons produced during pregnancy in a cohort from East African republic of Malawi. AB - Interferons (IFNs) associated with pregnancy were studied for their possible role in inhibition of vertical transmission of the human immunodeficiency virus type 1 (HIV-1). A study group was composed of 43 HIV-1-positive mothers, of whom 15 transmitted the virus to the offspring and 28 did not. The control group included 48 HIV-1-negative mother-infant pairs. The IFN-alpha was detected only sporadically in the maternal sera from the groups of transmitters (27%), nontransmitters (21%), and controls (19%). The average levels of IFN-alpha were low, 16.3 +/- 2.5 pg/ml, 21.4 +/- 9.9 pg/ml, and 21.3 +/- 9.4 pg/ml among the transmitters, nontransmitters, and control subjects, respectively. In the cord blood, IFN-alpha was detected only on two occasions among transmitters, and on a single occasion in the control group. IFN-beta was absent from both maternal and cord blood in the study group, and found to be present in one case in the control group simultaneously in the maternal and fetal sera. In the placentas, on the other hand, both type I and II IFNs were expressed universally in the villous trophoblast, and IFN-alpha and -beta in the stromal macrophages as well. In one case among transmitters, no IFNs were detected; nevertheless, no significant difference with respect to nontransmitters could be confirmed. Our data suggest that although the placental IFNs have an antiviral potential, they are not sufficient to suppress transmission of HIV from mother to infant. PMID- 10797375 TI - Seroprevalence of herpes simplex virus type 1 and type 2 in selected German populations-relevance for the incidence of genital herpes. AB - This study was carried out to determine the prevalence of antibodies to herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) in selected German populations, such as blood donors, hospital patients, and human immunodeficiency virus (HIV) seropositive individuals. Serum samples collected between 1996 and 1998 were tested by enzyme immunoassays using monoclonal antibody-selected native gG1 and gG2 as antigens and an immunoblot using type-specific recombinant glycoproteins. Equivocal results were resolved by an "in-house" Western blot assay. The prevalence of HSV-1 antibodies increased steadily with age and reached high levels of >/=88% among subjects 40 years of age or older. In the sample of patients and blood donors, the HSV-2 seroprevalence was 12.8% (95% CI = 11.9 13.8%). About 81% of the HSV-2 seropositive subjects were coinfected with HSV-1. When adjusted for age, there was no difference in the HSV-2 seroprevalence between hospital patients and blood donors. The HSV-2 seroprevalence was significantly higher among women (15%) than among men (10.5%), yielding a female : male odds ratio of 1.5 for hospital patients and of 1.67 for blood donors. Among the HIV-infected population, 91.1% were seropositive for HSV-1 and 47.9% for HSV-2. HIV-infected women have a significantly higher risk of HSV-2 infection than men (odds ratio [OR] = 3.22; 95% confidence ratio [CI] 1.99-5.20). In conclusion, although the rate of infections with HSV-2 is relatively low in the German population, attention should be given to the further development in adolescents, especially in view of a possible decrease of HSV-1 seroprevalence in childhood. PMID- 10797376 TI - Monitoring of human herpesvirus-6 and -7 genomes in saliva samples of healthy adults by competitive quantitative PCR. AB - Human herpesviruses-6 and -7 (HHV-6 and HHV-7) are thought to be transmitted during early infancy through saliva. However, the kinetics of the virus shedding in saliva of healthy adults, from whom children are assumed to acquire the viruses, is mostly unknown. This study was conducted to determine how many copies of the genome are secreted in saliva of healthy adults and to clarify the relationship between viral DNA load and virus isolation of HHV-6 and HHV-7. Competitive PCR was performed using primer sets in the U42 gene of each viral genome. In saliva samples from 29 healthy adults, HHV-6 and HHV-7 DNA was detected in 41.4% and 89.7%, respectively. The average copy number of the HHV-7 genome in the positive samples was higher than that of the HHV-6 genome. Follow up studies of six seropositive individuals for 3 months showed that the amount of HHV-7 DNA was constant in each individual and that "high producers" and "low producers" could be distinguished. By contrast, the amount of HHV-6 DNA varied drastically over time in each individual. Although HHV-6 was never isolated from the saliva of any of the six individuals during the follow-up period, HHV-7 was isolated from each individual several times. The amount of HHV-7 DNA tended to be higher at the times when the virus was isolated than at the times when the virus was not isolated. These data demonstrate a striking contrast between HHV-6 and HHV-7 in the kinetics of genome and virus shedding. PMID- 10797377 TI - Enterovirus RNA in serum is a risk factor for beta-cell autoimmunity and clinical type 1 diabetes: a prospective study. Childhood Diabetes in Finland (DiMe) Study Group. AB - Recent prospective studies have documented serologically an increased frequency of enterovirus infections in prediabetic children, indicating that these infections may initiate and accelerate the beta-cell damaging process several years before the clinical manifestation of type 1 diabetes. The aim of the present study was to establish whether these serological findings would be supported by the detection of enterovirus RNA in a unique prospective series of sera collected from prediabetic children 0-10 years before the manifestation of clinical type 1 diabetes. Reverse transcription followed by polymerase chain reaction employing highly conserved primers among enteroviruses were used to amplify enteroviral sequences. Viral RNA was found in 22% (11/49) of follow-up samples from prediabetic children but in only 2% (2/105) of those from controls (OR 14.9, P < 0.001). Persisting RNA positivity was not observed in any of these children. The presence of enterovirus RNA was associated with concomitant increases in the levels of autoantibodies against islet cells (OR 21.7, P < 0.01) and glutamic acid decarboxylase (OR 15.4, P < 0.05), but not in the levels of antibodies against insulin or the tyrosine phosphatase-like IA-2 protein. In contrast to the prediabetic children, those with newly diagnosed type 1 diabetes were negative for enterovirus RNA. The results thus complement previous serological data, suggesting that enterovirus infections are an important risk factor underlying type 1 diabetes and associated with the induction of beta-cell autoimmunity even years before symptoms appear. PMID- 10797378 TI - Evaluation of enterovirus serological tests IgM-EIA and complement fixation in patients with meningitis, confirmed by detection of enteroviral RNA by RT-PCR in cerebrospinal fluid. AB - An enzyme immunoassay (EIA) for detection of anti-enterovirus IgM antibodies was compared with complement fixation test in 43 patients with confirmed enterovirus meningitis by RT-PCR of cerebrospinal fluids (CSF). In 34% of patients with enterovirus meningitis, IgM antibodies could be found, whereas complement fixation tests were positive in only 20%. The specificity was determined with sera of 105 patients with non-enterovirus meningitis. Specificity of IgM EIA and of complement fixation was 94% and 85%, respectively. In four patients with meningitis but without enterovirus detection in CSF, RT-PCR and virus isolation from stools were positive. In three of these patients, IgM antibodies were detected, giving a strong indication of an enterovirus-associated disease. Because of the high specificity of IgM EIA, diagnosis of enterovirus-associated diseases can be carried out in a single serum sample, whereas by complement fixation tests, only fourfold increases in antibody titres in paired sera indicate an acute infection. The application of IgM EIA is especially important in cases of meningitis when CSF samples are not available and for diagnosis of enterovirus diseases with other clinical symptoms such as fever, enteritis, and hand-foot-and-mouth disease. PMID- 10797379 TI - Expression of capsid [correction of caspid] protein VP1 for use as antigen for the diagnosis of enterovirus 71 infection. AB - To produce enterovirus 71 antigen for diagnostic purposes, the gene encoding the entire capsid protein VP1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned and expressed in Escherichia coli as a poly-histidine fusion protein. Western blotting experiments with sera from patients with enterovirus 71 infection indicated that immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide VP1. According to these results, IgM anti-VP1 appeared in sera of patients with a symptomatic enterovirus 71 acute infection, whereas IgG anti-VP1 was present in sera of past infection. This finding suggests that detecting IgG and IgM immune responses against linear epitopes of recombinant VP1 is an effective means of determining the different phases of enterovirus 71 infection. In addition, sera containing coxsackie virus 16 (CA16) antibodies did not cross-react with the recombinant VP1 of enterovirus 71, despite the homology between VP1 proteins of both viruses. Comparison with reference PCR and neutralization assays showed these antibody tests to be appropriate for the serodiagnosis of enterovirus 71 infection. PMID- 10797380 TI - Epstein-Barr virus reactivation associated with diminished cell-mediated immunity in antarctic expeditioners. AB - Epstein-Barr virus (EBV) reactivation and cell-mediated immune (CMI) responses were followed in 16 Antarctic expeditioners during winter-over isolation at 2 Australian National Antarctic Research Expedition stations. Delayed-type hypersensitivity (DTH) skin testing was used as an indicator of the CMI response, that was evaluated 2 times before winter isolation and 3 times during isolation. At all 5 evaluation times, 8 or more of the 16 subjects had a diminished CMI response. Diminished DTH was observed on every test occasion in 4/16 subjects; only 2/16 subjects exhibited normal DTH responses for all 5 tests. A polymerase chain reaction (PCR) assay was used to detect EBV DNA in saliva specimens collected before, during, and after the winter isolation. EBV DNA was present in 17% (111/642) of the saliva specimens; all 16 subjects shed EBV in their saliva on at least 1 occasion. The probability of EBV shedding increased (P = 0.013) from 6% before or after winter isolation to 13% during the winter period. EBV appeared in saliva during the winter isolation more frequently (P < 0.0005) when DTH response was diminished than when DTH was normal. The findings indicate that the psychosocial, physical, and other stresses associated with working and living in physical isolation during the Antarctic winter result in diminished CMI and an accompanying increased reactivation and shedding of latent viruses. PMID- 10797381 TI - Expression of the Epstein-Barr virus BHRF1 gene, a homologue of Bcl-2, in nasopharyngeal carcinoma tissue. AB - Epstein-Barr virus (EBV) infection is associated closely with the pathogenesis of nasopharyngeal carcinoma (NPC). The EBV gene product, BHRF1, has been demonstrated in vitro and is structurally and functionally similar to the oncogene bcl-2, that is able to protect cells from programmed cell death. To determine whether the BHRF1 gene is expressed in vivo, BHRF1 mRNA or protein were sought in tissues from NPC and non-NPC patients. BHRF1 transcripts were specifically detected in the NPC tumours (32 out of 44, 72.7%) rather than the non-NPC tissues (0 out of 25) by reverse transcription, polymerase chain reaction and Southern hybridization. Other EBV genes, such as the lytic gene BZLF1 and latent genes EBNA1 and LMP2A, were also investigated. BZLF1 transcripts also were found specifically in NPC tumours (33 out of 44, 75%). EBNA1 was expressed in 79.5% of NPC, and 28% of non- NPC, tissues and LMP2A was expressed in 70.5% of NPC, and 88% of non-NPC, tissues. BHRF1 protein was detected by immunohistochemistry in 4 metastatic NPC, of 36 NPC tissue sections available. The BHRF1 protein was distributed in both the nucleus and cytoplasm of the neoplastic epithelial cells. IgG antibody against the BHRF1 protein was detected in 6 of 17 (35. 3%) NPC plasma, but the protein and IgG were both absent from the non-NPC controls. BHRF1 DNA sequences were determined for 11 NPC and 3 non-NPC samples. No sequence was specific for the EBV isolates from NPC tissue. Amino acids 79 and 88 always appeared in the same form, however, for every tested isolate and both were valine or leucine. This particular characteristic was not present in the B95-8 strain or in the corresponding regions of homologues, Bcl-2 and Bcl-XL, and was regarded as unique to Oriental EBV strains. PMID- 10797382 TI - Multiplex polymerase chain reaction for the evaluation of cytomegalovirus DNA load in organ transplant recipients. AB - Because of the considerable impact of human cytomegalovirus (HCMV) infection, sensitive, specific, and standardized methods are required for rapid and accurate evaluation of viral load in monitoring transplant recipients. The aim of the present study was to evaluate the usefulness of a multiplex polymerase chain reaction (PCR) for the coamplification of HCMV-DNA and beta-globin genomic sequence in polymorphonuclear leukocytes (PMNL). Analysis and quantification of PCR products were carried out by a DNA enzyme immunoassay (DEIA), which is based on the hybridization of amplified DNA with a single-stranded DNA probe, which coats microtitre wells. Colorimetric detection of the DNA-antibody complex was carried out and optical density (O.D.) was recorded at 450/630 nm. To quantify HCMV/DNA load, a standard curve to which samples O.D. refer was obtained by amplifying serial dilutions of recombinant PGEM-3Z plasmid DNA containing a genomic fragment of glycoprotein B. 340 PMNL specimens from 102 solid organ recipients were tested for the detection of pp65 antigen and HCMV-DNA. The results showed a good correlation between viral load and clinical symptoms of HCMV infection; high specificity and predictive values for HCMV disease were found by PCR, using a cut-off limit of 10(3) genomic copies per 2 x 10(5) PMNL. These findings indicate that the system described is an efficient and reproducible diagnostic method easy to apply for routine diagnosis and therapeutic monitoring of transplanted patients. PMID- 10797383 TI - Immunisation with gamma globulin to murray valley encephalitis virus and with an inactivated Japanese encephalitis virus vaccine as prophylaxis against australian encephalitis: evaluation in a mouse model. AB - In northwestern Australia, the flavivirus Murray Valley encephalitis (MVE) poses a significant health risk to infants in some aboriginal communities, particularly during each wet season. While there are too few cases to warrant the development of a vaccine against MVE, a safe, effective prophylaxis for these children is still urgently required. The use of passive transfer of human gamma globulin to MVE or immunisation with a vaccine to the closely related Japanese encephalitis (JE) virus were investigated as potential strategies. When 40 microg of IgG was purified from MVE-immune human sera and transferred to 3-week-old mice, the animals were protected from lethal IP inoculation with MVE virus while still producing a detectable immune response to the virus. Similarly, sera from adult mice infected sublethally with MVE or JE virus provided significant protection against MVE infection. However, sera from mice sublethally infected with the related Kunjin or immunised with the inactivated JE vaccine (Biken) provided no protection against MVE challenge. In fact, mice immunised passively with the latter appeared to succumb to MVE challenge more rapidly than mice that received serum from unimmunised animals, suggesting that antibody to the vaccine had accelerated the progression of disease. These preliminary trials in mice indicate that passive immunisation with human gamma globulin has the greatest potential as a strategy for MVE prophylaxis, whilst the apparent enhancement of MVE by antibodies to the JE vaccine requires further investigation, with particular reference to current vaccination programs in areas of Australia and Papua New Guinea, where both JE and MVE occur. PMID- 10797384 TI - Antigenic properties and diagnostic potential of recombinant dobrava virus nucleocapsid protein. AB - Dobrava hantavirus (DOBV) causes severe hemorrhagic fever with renal syndrome in the Balkan region and has been detected recently also in Russia, Estonia, and Germany. DOBV nucleocapsid protein (N) was produced in insect cells, using the baculovirus expression system (bac-DOBV-N), and in E. coli as a truncated (aa 1 165) glutathione-S transferase fusion protein (DOBV-dN-GST). The antigenic properties of bac-DOBV-N were found identical to native DOBV-N when examined by a panel of hantavirus-specific monoclonal antibodies. Enzyme immunoassays for detection of IgM and IgG antibodies were set up using DOBV recombinant N proteins and compared with those based on recombinant Hantaan and Puumala virus N, using panels of sera collected from DOBV, Hantaan and Puumala virus-infected patients. Full-length N protein (bac-DOBV-N) was found to be a more sensitive antigen than DOBV-dN-GST. The sensitivity values for sera from DOBV-infected patients were 100% for bac-DOBV-N and 86% for DOBV-dN-GST by IgM assays, and 98% for bac-DOBV-N and 88% for DOBV-dN-GST by IgG assays. The specificity values were 100% for bac DOBV-N and 99% for DOBV-dN-GST by IgM assays, and 100% for both antigens by IgG assays. PMID- 10797385 TI - Respiratory syncytial virus: changes in prevalence of subgroups A and B among Argentinian children, 1990-1996. AB - The frequency of respiratory syncytial virus (RSV) and the distribution of subgroups A and B strains during 7 consecutive years (1990-1996) were examined in two cities of Argentina. Nasopharyngeal aspirates from 1,304 children less than 2 years of age hospitalized with acute lower respiratory infection were studied by indirect immunofluorescence. RSV was detected in 352 cases (26.9%), and the peak activity was observed in midwinter. Subgroup characterization was performed with two monoclonal antibodies against the F protein on nasopharyngeal aspirate smears. Of 195 samples, 174 (89.2%) were identified as subgroup A strains and 21 (10.8%) as subgroup B. Both strains cocirculated during 5 of 7 years studied with subgroup A predominating. Subgroup A occurred at least 8 times as often in all years except for 1994-1995. Children infected by subgroup A were younger than those infected by subgroup B (P < 0.05). The association of subgroup A infection with bronchiolitis and subgroup B with pneumonia was statistically significant (P < 0.03). PMID- 10797387 TI - Understanding neuromyotonia. PMID- 10797388 TI - Motor neuron disease and malignancy. PMID- 10797389 TI - Myosin heavy chain isoform expression following reduced neuromuscular activity: potential regulatory mechanisms. AB - In this review, the adaptations in myosin heavy chain (MHC) isoform expression induced by chronic reductions in neuromuscular activity (including electrical activation and load bearing) of the intact neuromuscular unit are summarized and evaluated. Several different animal models and human clinical conditions of reduced neuromuscular activity are categorized based on the manner and extent to which they alter the levels of electrical activation and load bearing, resulting in three main categories of reduced activity. These are: 1) reduced activation and load bearing (including spinal cord injury, spinal cord transection, and limb immobilization with the muscle in a shortened position); 2) reduced loading (including spaceflight, hindlimb unloading, bed rest, and unilateral limb unloading); and 3) inactivity (including spinal cord isolation and blockage of motoneuron action potential conduction by tetrodotoxin). All of the models discussed resulted in increased expression of fast MHC isoforms at the protein and/or mRNA levels in slow and fast muscles (with the possible exception of unilateral limb unloading in humans). However, the specific fast MHC isoforms that are induced (usually the MHC-IIx isoform in slow muscle and the MHC-IIb isoform in fast muscle) and the degree and rate of adaptation are dependent upon the animal species and the specific model or condition that is being studied. Recent studies designed to elucidate the mechanisms by which electrical activation and load bearing alter expression of MHC isoforms at the cellular and genetic levels are also reviewed. Two main mechanisms have been proposed, the myogenin:MyoD and calcineurin:NF-AT pathways. Collectively, the data suggest that the regulation of MHC isoform expression involves a complex interaction of multiple control mechanisms including the myogenin:MyoD and calcineurin:NF-AT pathways; however, other intracellular signaling pathways are likely to contribute. PMID- 10797390 TI - Surgery for peripheral nerve and brachial plexus injuries or other nerve lesions. AB - The selection of patients who will benefit from peripheral nerve surgery continues to evolve. The evaluation of patients with peripheral nerve problems, and the indications and timing of surgery are reviewed. This includes patients with transections, lesions in continuity, entrapments, tumors, injection injuries, and birth palsies. The treatment outlined for patients with peripheral nerve and brachial plexus injuries is based on a comprehensive clinical and electrodiagnostic evaluation complemented by imaging studies. A systematic approach to the surgical management of these neural lesions has evolved at our institution, based in part on extensive experience with intraoperative recording of nerve action potentials (NAPs) and a resultant large number of referrals. Advanced microsurgical techniques including use of grafts have expanded the scope of peripheral nerve surgery. These advances coupled with intraoperative electrophysiologic measurements have improved outcomes. PMID- 10797391 TI - Neuromyotonia in mice with hereditary myelinopathies. AB - The purpose of this study was to further characterize neuromyotonia in mice with deletions and point mutations of myelin protein genes. Clinical observation showed irregular stretching of the hindlimbs, tremor and generalized myokymia in mice with targeted deletions of the genes encoding myelin protein zero (P0-/-) or peripheral myelin protein 22 (Pmp22-/-), and Trembler mice, which carry a point mutation of Pmp22. By electromyography (EMG), we found irregular high-frequency bursts of spontaneous motor unit activity and rhythmic doublet or multiplet discharges of motor units in these mouse models of human hereditary neuropathies. The EMG signs are typical for neuromyotonia and myokymia, respectively. The activity persisted after a proximal nerve section in many cases, localizing the generator to the peripheral nerve or the muscle. We now show that blocking neuromuscular transmission with suxamethonium abolished the spontaneous activity, ruling out a muscle origin. Phenytoin ameliorated the motor behavior. Taken together, our study shows that neuromyotonia develops in different mouse models of hereditary myelinopathies. This indicates that spontaneous motor unit activity may underlie neuromyotonia, which is occasionally observed in Charcot-Marie-Tooth disease. These animal models will be useful to study the pathogenesis of neuromyotonia. PMID- 10797392 TI - Chronic motor axonal neuropathy associated with antibodies monospecific for N acetylgalactosaminyl GD1a. AB - We report on three patients with chronic motor neuropathy who had elevated titers of immunoglobulin (Ig)G antibodies against N-acetylgalactosaminyl GD1a (GalNAc GD1a) and normal titers of antibodies against other gangliosides. Presenting with progressive muscular atrophy, fasciculations, and no sensory deficits, the patients had been diagnosed to have motor neuron disease. Electrodiagnostic features were predominantly axonal. Two patients clinically improved after intravenous Ig infusion and cyclophosphamide therapy. Increased titers of IgM antibodies to GalNAc-GD1a were also found in two of 15 patients with multifocal motor neuropathy with conduction block but were associated with concomitant rise of anti-GM1 antibodies. These three cases represent a chronic motor axonal neuropathy in which antibody testing for a minor ganglioside was helpful for instituting therapy. PMID- 10797393 TI - Skeletal muscle recovery after tenotomy and 7-day delayed muscle length restoration. AB - Rabbit extensor digitorum longus (EDL) tendons were cut with the muscle active (active tenotomy, AT) or with the EDL at rest (passive tenotomy, PT). One, 7, and 21 days after tenotomy, contractile testing was performed. A second experiment was performed in which EDL tendons underwent PT and, after a 7-day delay, muscle tendon units were restored to their original length. Maximum isometric tension dropped precipitously 1 day after either AT or PT to approximately 50% of normal and continued to decline by day 7. In contrast to PT, where peak tension (P(0)) decreased further by 21 days, after AT, P(0) partially recovered. Differences in muscle mass, cross-sectional area, fiber type, and sarcomere number did not explain the differential response. One day after length restoration of muscles, P(0) rapidly increased by approximately 40%. These observations have implications for understanding the outcome of muscle-tendon unit injury and surgical repair. PMID- 10797394 TI - Concentric needle electrode for neuromuscular jitter analysis. AB - We used a concentric needle electrode (CNE) with 2 kHZ low-cut filter and a single fiber electrode (SFE) in the same subjects for neuromuscular jitter measurement in the extensor digitorum communis (EDC) and orbicularis oculi (OOc) muscles. At the same session, 20 jitter values were obtained from each subject with each electrode. For EDC (during voluntary contraction), mean jitter values with SFE and CNE were 23.4 +/- 8 micros and 23.3 +/- 8 micros in 10 normals; and 56.8 +/- 28 micros and 57.4 +/- 33 micros in 10 myasthenics. For OOc (during electrical stimulation), mean jitter values with SFE and CNE were 17.9 +/- 5 micros and 16.3 +/- 4 micros in 11 normal subjects, and 41.2 +/- 29 micros and 36.7 +/- 27 micros in 10 myasthenics. For both muscles, the numbers of individual abnormal jitter values with SFE and CNE were highly comparable. Both needles labeled the same patients as having "normal" or "abnormal" neuromuscular transmission. CNE may be an alternative to SFE in neuromuscular jitter analysis. PMID- 10797396 TI - Anal sphincter EMG does not distinguish between multiple system atrophy and Parkinson's disease. AB - Clinical distinction of multiple system atrophy (MSA) from Parkinson's disease (PD) is often difficult. Several recent reports indicate that objective classification may be accomplished using electromyographic (EMG) testing of the anal or urethral sphincters, but some authors have found that these tests are not reliable for this purpose. We studied 13 patients with PD and 10 with probable MSA, as diagnosed by consensus of four movement disorders specialists, according to accepted clinical criteria. Anal sphincter EMG was performed blind to the clinical diagnosis. We found no significant differences in the mean duration of motor unit potentials (MUPs), mean MUP amplitude, or prevalence of polyphasic potentials, satellite potentials, very long duration MUPs, or spontaneous activity between the two groups. Thus, anal sphincter EMG does not differentiate between PD and MSA. PMID- 10797395 TI - Rabies virus entry at the neuromuscular junction in nerve-muscle cocultures. AB - Early events in rabies virus entry into neurons were investigated in chick spinal cord-muscle cocultures. Rabies virus (CVS strain) was adsorbed to the surface of cells in the cold. At times up to 10 min of warming to 37 degrees C, virus was most intensely localized to dense swellings on the myotube surface. Texas Red labeled alpha-bungarotoxin, which binds to nicotinic acetylcholine receptors, colocalized precisely with virus at the densities identifying these regions as neuromuscular junctions. Rabies virus also colocalized in the junctions with synapsin I, a marker for synaptic vesicles. The endosome tracers Lucifer Yellow, Texan Red-dextran, and rhodamine-wheat germ agglutinin were added to the cultures at the end of the virus adsorption period and the cultures were warmed. At 10 min, rabies virus and tracers colocalized at neuromuscular junctions and nerve terminals. At 30 min, rabies virus and tracers showed more intense fluorescence over nerve fibers and nerve cell bodies. At 60 min, nerve terminals, nerve fibers, and nerve cell bodies showed intense fluorescence and colocalization for rabies virus and tracers. LysoTracker Red, a marker for acidic compartments, colocalized with rabies virus at nerve-muscle contacts. These findings show that in nerve-muscle cocultures, the neuromuscular junction is the major site of entry into neurons. Colocalization of virus and endosome tracers within nerve terminals indicates that virus resides in an early endosome compartment, some of which are acidified. The progressive increase of virus and tracers in nerve fibers and nerve cell bodies over time is consistent with retrograde transport of endocytosed virus from the motor nerve terminal. PMID- 10797397 TI - Persistence of myosin heavy chain-based fiber types in innervated but silenced rat fast muscle. AB - Myosin heavy chain (MHC) profile and size of fibers in deep and superficial regions of the adult rat medial gastrocnemius (MG) were determined after 4, 15, 30, and 60 days of inactivity induced by spinal cord isolation (SI). After 4 days, fiber size decreased by 33 to 50% and 36 to 46% in deep and superficial regions, whereas MHC composition was unaffected. By 15 days, these values were 45 to 78% and 51 to 69%, and MHC composition was shifting toward faster isoforms. By 60 days, there were no pure type I MHC fibers and increases from 1 to 18% and 78 to 93% in pure type IIb fibers in deep and superficial regions. The percentage of type I MHC (gel electrophoresis) was approximately 10 and approximately 3%, and of type IIb approximately 40 and approximately 60% in control and 60-day SI rats. Thus, adaptations in the MHC molecule occurred at a slower rate and for a longer duration than the atrophic response. PMID- 10797399 TI - Comparison of the response of motoneurons innervating perineal and hind limb muscles to spaceflight and recovery. AB - The succinate dehydrogenase (SDH) activities and cell body sizes of motoneurons in the dorsomedial (DM) region of the ventral horn at the lower portion of the L5 and the L6 segmental levels of the rat spinal cord were determined following 14 days of spaceflight and after 9 days of recovery on Earth and compared with those in the retrodorsolateral (RDL) region of the ventral horn at the same segmental levels. No changes in the mean SDH activity of motoneurons in the DM region were observed following spaceflight or after recovery. However, a decrease in the mean SDH activity of motoneurons with cell body sizes between 500 and 900 microm(2) in the RDL region was observed following spaceflight and after recovery. These data indicate that moderate-sized motoneurons in the RDL region, which are most likely associated with the hind limb musculature, were responsive to the microgravity environment. In contrast, the motoneurons in the DM region associated with the perineal muscles (associated with predominantly fast, low-oxidative muscles which are recruited for relatively brief periods at high activation levels and have no load-bearing function at 1G) were not affected by microgravity. PMID- 10797398 TI - Ia presynaptic inhibition after muscle twitch in the arm. AB - Contraction of upper limb muscles in healthy subjects was used to investigate presynaptic inhibition at spinal level. The H reflex recorded in the forearm flexor muscles in response to median nerve stimulation was depressed in amplitude from 400 ms to 1 s after a muscle twitch induced by transcranial stimulation, root stimulation, direct biceps stimulation, and triceps tendon tap. Stimulation of the cutaneous branch of musculocutaneous nerve, ipsilateral triceps and contralateral biceps, and biceps tendon tap did not alter H-reflex size. Forearm flexor H-reflex amplitude is therefore related to changes in proprioceptive inflow secondary to the biceps muscle twitch. Root and direct muscle stimulation both failed to reduce the size of the motor evoked potential (MEP) after transcranial magnetic stimulation, suggesting that the inhibition acts at presynaptic level. Attenuation of H-reflex amplitude was related to the size of the muscle twitch and was less pronounced during an isometric twitch than during free joint movement. Our results suggest that the biceps muscle twitch produces long-lasting inhibition of the Ia afferents from forearm flexor muscles. This is an important and a simple mechanism for suppressing proprioceptive input during movement. PMID- 10797400 TI - Recovery of excitability of cutaneous afferents in the median and sural nerves following activity. AB - In acquired polyneuropathies, symptoms and signs are typically distal and symmetrical, more prominent in the lower limbs than the upper limbs. This study was undertaken to measure the extent of the decrease in excitability produced by single impulses and by impulse trains in cutaneous afferents in the median and sural nerves, and to compare the resulting changes in excitability of these afferents. Threshold tracking was used in 10 healthy subjects to measure the changes in threshold for a compound sensory action potential of 50% maximum produced by conditioning stimuli. Following a single supramaximal conditioning stimulus, the threshold changes occurring during the refractory and supernormal periods were identical for the two nerves, but there was a greater increase in threshold during the late subnormal period for median afferents. Following a train of 10 supramaximal conditioning stimuli, threshold increased by approximately 40% for median afferents and by approximately 20% for sural afferents. These differences are consistent with differences in a slow K(+) conductance. It is suggested that the hypo-excitability produced by brief trains of impulses may be sufficient to disturb conduction in diseased nerve fibers, and that the lesser expression of slow K(+) conductances on cutaneous afferents in the sural nerve could render them more sensitive to depolarizing stresses than median afferents. This could be a factor in the ease with which sural afferents become ectopically active in polyneuropathies. PMID- 10797401 TI - Strength training can improve steadiness in persons with essential tremor. AB - We evaluated the effect of a strength-training program on the ability of persons with essential tremor to exert steady forces with the index finger. Thirteen subjects with a diagnosis of essential tremor were assigned to three different groups: one group trained with heavy loads, one with light loads, and one did not perform any training. Subjects attempted to generate steady contractions during both postural and constant-force tasks. Steadiness was quantified by the root mean square amplitude of acceleration during postural tasks and the standard deviation and coefficient of variation of force during the constant-force tasks. Subjects who performed the training program with heavy loads experienced an increase in steadiness around the target force during the constant-force tasks. Subjects in the other two groups did not exhibit any changes. These findings suggest that strength training can decrease the magnitude of tremor. However, we did not observe any associated improvements in functional abilities. PMID- 10797402 TI - Stimulated single fiber EMG of the frontalis muscle in the diagnosis of ocular myasthenia. AB - We performed single fiber electromyography by axonal stimulation (SFEMG-AS) of the frontalis muscle of 16 patients with ocular myasthenia gravis (OM) and 33 controls. In the controls, values of mean consecutive differences (MCD) ranged from 5 to 55 micros (average, 14.7 +/- 2.8 micros) and mean MCD of individual MPs was 14. 6 +/- 6.8 micros. All the OM patients showed abnormal SFEMG-AS jitter before prostigmine was administered (mean MCD: 49.19 +/- 21. 82 micros, percentage of blocks: 20.97 +/- 18.53). Twenty or 30 min after prostigmine had been administered, we saw a significant improvement in jitter: mean MCD was 36.38 +/- 22.49 micros (P = 0. 005), and percentage of blocks was 10.16 +/- 18.87 (P = 0.008). The method was well tolerated. We conclude that SFEMG-AS of the frontalis muscle is a sensitive technique for the diagnosis of OM and is easy to carry out. PMID- 10797403 TI - Dystrophin mutations predict cellular susceptibility to oxidative stress. AB - Mutations in the dystrophin gene that lead to the expression of truncated forms of the dystrophin protein cause muscular dystrophies of varying severities both in humans and in mice. We have shown previously that dystrophin-deficient muscle is more susceptible to oxidative injury than is normal muscle. In this report, we have used muscle cells derived from mdx mice, which express no dystrophin, and mdx-transgenic strains that express full-length dystrophin or truncated forms of dystrophin to explore further the relationship between dystrophin expression and susceptibility of muscle to oxidative injury. We show that, when differentiated into myotubes, the relative susceptibility of the cell populations to oxidative stress correlates with the severity of the dystrophy in the strain from which the cells were isolated. The most susceptible populations exhibited the greatest oxidative damage as assessed by protein oxidation. Thus, the relative efficacy of truncated dystrophin proteins to protect muscle from necrotic degeneration in vivo is predicted by their ability to protect muscle cells from free radical mediated injury. These findings support the hypothesis that the dystrophin protein complex may have important regulatory or signaling properties in terms of cell survival and antioxidant defense mechanisms. PMID- 10797404 TI - Flexor reflexes in chronic spinal cord injury triggered by imposed ankle rotation. AB - Hypersensitivity of the flexor reflexes to input from force-sensitive muscle afferents may contribute to the prevalence and severity of muscle spasms in patients with spinal cord injuries. In the present study, we triggered flexor reflexes with constant-velocity ankle movements into end-range dorsiflexion and plantarflexion positions in 8 individuals with spinal cord injuries. We found that all 8 subjects had coordinated increases in flexion torque at the hip and ankle following externally imposed plantarflexion movements at the ankle. In addition, end-range dorsiflexion movements also triggered flexor reflexes in 3 subjects, although greater loads were required to trigger such reflexes using dorsiflexion movements (compared to plantarflexion movements). These three-joint reflex torque patterns triggered by ankle movement were broadly comparable to flexion withdrawal responses elicited by electrocutaneous stimuli applied to a toe, although the amplitude of the torque response was generally lower. We conclude that excitation of muscle and joint-related afferents induced by end range movements may be responsible for exaggerated flexion reflex responses in spinal cord injury. PMID- 10797405 TI - CTG triplet repeat expansion in a laryngeal carcinoma from a patient with myotonic dystrophy. AB - A 66-year-old Japanese man with myotonic dystrophy (DM) underwent total laryngectomy for laryngeal carcinoma. The size of the expanded DNA fragment (EF) from the leukocytes and normal laryngeal tissues of this patient was only slightly longer than that in normal subjects. EF, however, was markedly longer in the laryngeal carcinoma. These findings support the hypothesis that elongation of the CTG repeat in the DM kinase gene occurs during acquired cell proliferation. PMID- 10797406 TI - Intrafamilial phenotypic variation in limb-girdle muscular dystrophy type 2C with compound heterozygous mutations. AB - Two Japanese-Brazilian siblings with type 2C limb girdle muscular dystrophy showed a maternal 521-T deletion in exon 6 and a larger paternal deletion of exon 6 in the gamma-sarcoglycan gene. One sib was ambulant at 29 years of age, whereas the other sib was confined to a wheelchair at the age of 12. Sarcoglycan staining of the muscle was reduced in both siblings but it did not correlate with the observed variability of the clinical severity. PMID- 10797407 TI - 1999 scientific session of the Myasthenia Gravis Foundation. PMID- 10797408 TI - Delayed radiation-induced bulbar palsy mimicking ALS. AB - We describe a patient presenting with progressive bulbar dysfunction and spasticity that clinically mimicked amyotrophic lateral sclerosis (ALS). Electromyography, however, showed no evidence of denervation and revealed a rare combination of peripheral and central myokymia. We feel that this pattern of myokymia represented a marker of neural injury from remote radiation therapy. Nervous system disorders resulting from therapeutic radiation are described, and potential pathophysiologic mechanisms underlying myokymia are discussed. PMID- 10797409 TI - Becker muscular dystrophy combined with X-linked Charcot-Marie-Tooth neuropathy. AB - A man was identified with two X-chromosomal neuromuscular disorders, X-linked Charcot-Marie-Tooth disease (CMTX) and Becker muscular dystrophy (BMD). The neuropathy could be tracked in the family and was found to be caused by a mutation in the connexin32 gene on Xq13. 1. The muscular dystrophy was sporadic owing to a de novo deletion in the dystrophin gene located in band Xp21.2. Although these genetic alterations of the same X-chromosome are considered as physically independent, their combination resulted in a unique phenotype with severe wasting of proximal as well as distal muscles and rapid progression of both conditions. PMID- 10797410 TI - Polymyalgia rheumatica following influenza vaccination. PMID- 10797411 TI - Diabetic muscular infarction. PMID- 10797412 TI - Slow repolarization phase of the intracellular action potential influences the motor unit action potential. PMID- 10797413 TI - Reply. PMID- 10797414 TI - AAEM news and comments. PMID- 10797415 TI - Asymptomatic maternal myasthenia as a cause of the Pena-Shokeir phenotype. AB - We report six sibs with arthrogryposis multiplex congenita and a Pena-Shokeir phenotype, born to a healthy woman who was discovered to have asymptomatic myasthenia gravis (MG). This is the first report of anti-acetylcholine receptor (AChR) antibodies causing fetal akinesia/hypokinesia sequence in the offspring of an asymptomatic mother. PMID- 10797416 TI - Two novel fibrillin-2 mutations in congenital contractural arachnodactyly. AB - Congenital contractural arachnodactyly (CCA) is an autosomal dominant connective tissue disorder, comprising marfanoid habitus, flexion contractures, severe kyphoscoliosis, abnormal pinnae, and muscular hypoplasia. It is now known that mutations in the gene encoding fibrillin-2 cause CCA. Interestingly, mutations described to date cluster in the fibrillin-2 region homologous to the so-called neonatal Marfan syndrome region of fibrillin-1. Thus, it has been hypothesized that the relative infrequency of CCA compared with the Marfan syndrome is due to the limited region of the gene targeted for mutations. In support of the above hypothesis, we report here the finding of two additional FBN2 mutations in CCA, C1141F (exon 26) and C1252W (exon 29). In addition, a new 3' UTR polymorphism is also described. PMID- 10797417 TI - Body stalk defects, body wall defects, amniotic bands with and without body wall defects, and gastroschisis: comparative epidemiology. AB - The presence of body wall defects with "evisceration" of thoracic and/or abdominal organs associated with other congenital anomalies, with or without limb deficiencies, is considered to be the body wall complex (BWC). The BWC is different from gastroschisis, which is usually a small body wall defect lateral to the umbilical cord that is not covered by any membrane and, in most of the cases, is an isolated defect. For the present analysis we separated the BWC group into three subgroups. One group was that of body stalk anomalies characterized by severe defects of the abdominal wall with absence of, or very small, umbilical cord, or this is continuing with the placenta. The second group was made up of those infants with body wall defects without amniotic bands, and the third group was of those children with body wall defects produced by amniotic bands. We considered two additional groups in the analysis, one was of infants with gastroschisis and the other those infants with amniotic bands without body wall affectation. We also included the control group (nonmalformed infants) for comparisons. From the results of our epidemiological study, we can conclude that amniotic bands with body wall affectation and amniotic bands without body wall defects are two different entities. The results also suggest that the characteristics of infants with amniotic bands with body wall defects are more similar to the group of infants with body stalk anomalies. This may indicate that the former group is produced during the very early gestation. PMID- 10797418 TI - Submicroscopic deletion in cousins with Prader-Willi syndrome causes a grandmatrilineal inheritance pattern: effects of imprinting. AB - The Prader-Willi syndrome (PWS) critical region on 15q11-q13 is subject to imprinting. PWS becomes apparent when genes on the paternally inherited chromosome are not expressed. Familial PWS is rare. We report on a family in which a male and a female paternal first cousin both have PWS with cytogenetically normal karyotypes. Fluorescence in situ hybridization (FISH) analysis shows a submicroscopic deletion of SNRPN, but not the closely associated loci D15S10, D15S11, D15S63, and GABRB3. The cousins' fathers and two paternal aunts have the same deletion and are clinically normal. The grandmother of the cousins is deceased and not available for study, and their grandfather is not deleted for SNRPN. DNA methylation analysis of D15S63 is consistent with an abnormality of the imprinting center associated with PWS. "Grandmatrilineal" inheritance occurs when a woman with deletion of an imprinted, paternally expressed gene is at risk of having affected grandchildren through her sons. In this case, PWS does not become evident as long as the deletion is passed through the matrilineal line. This represents a unique inheritance pattern due to imprinting. PMID- 10797419 TI - Attitudes toward the genetic testing of children among adults in a Utah-based kindred tested for a BRCA1 mutation. AB - Advances in molecular biology and genetics have led to the identification of the breast/ovarian cancer susceptibility genes BRCA1 and BRCA2, along with tests to detect mutations in these genes. Although the appropriateness of BRCA1/2 genetic testing for children has been debated in the literature, little is known about the attitudes of individuals who have undergone cancer susceptibility testing. The present study focused on attitudes toward BRCA1 testing for children among 218 adults from a Utah-based kindred who had received BRCA1 test results. Results indicated that approximately one-fourth of the participants would permit BRCA1 testing for children under the age of 18. General attitudes about genetic testing were predictive of attitudes toward the testing of children. In addition, men and individuals without a BRCA1 mutation were more likely to agree that minors should be allowed BRCA1 testing. Individuals whose mother had been affected with breast cancer were less likely to permit testing for minors. Among parents of minor children, less than one-fifth indicated that they would want BRCA1 testing for their own children; carrier status was not predictive of attitudes toward testing their own children. As breast/ovarian cancer susceptibility testing continues to be disseminated into clinical settings, there may be an increase in the number of test requests for minors. The findings of the present study represent an important step in exploring attitudes about genetic testing of children among individuals who have received cancer susceptibility test results. PMID- 10797420 TI - SPONASTRIME dysplasia: report of an 11-year-old boy and review of the literature. AB - SPONASTRIME (SPOndylar and NAsal changes, with STRIations of the MEtaphyses) dysplasia is a rare, autosomal recessive bone disorder first described by Fanconi et al. [1983: Helv Paediatr Acta 38:267-280]. Radiographic findings include abnormal vertebral bodies with age-dependent changes, and striations of the metaphyses, scoliosis, and retarded ossification of the carpal bones. Physical features include severe short stature, lumbar lordosis, midface hypoplasia, frontal bossing, and a depressed nasal root. To date, 12 patients from 6 families have been reported. Four additional patients have been reported with a variant of this condition, which includes mental retardation. We report on an 11-year-old boy with features consistent with SPONASTRIME dysplasia. Height was 106.1 cm (-6 SD). He had a coarse appearing face with a depressed nasal bridge, short, upturned nose, and midface hypoplasia. Intelligence was normal. A clinical evaluation at 6 years of age suggested the diagnosis of spondyloepiphyseal dysplasia (SED). However, genetics evaluation at 11 years of age with repeat radiologic studies revealed delayed carpal ossification (-4 to -5 SD), metaphyseal irregularities and striations most notably in the distal femurs and the proximal tibias, lumbar lordosis, narrow interpedicular distances of the lumbar spine, and pear-shaped vertebral bodies. These findings were most consistent with the diagnosis of SPONASTRIME dysplasia, and not SED. Although radiographic findings of SPONASTRIME dysplasia are distinguishable from SED, the physical appearance may be similar. Many bone dysplasias have overlapping radiographic findings and clinical presentation but with different recurrence risks, making genetic counseling a challenge. PMID- 10797421 TI - Spectrum and detection rate of L1CAM mutations in isolated and familial cases with clinically suspected L1-disease. AB - Mutations in L1CAM, the gene encoding the L1 neuronal cell adhesion molecule, lead to an X-linked trait characterized by one or more of the symptoms of hydrocephalus, adducted thumbs, agenesis or hypoplasia of corpus callosum, spastic paraplegia, and mental retardation (L1-disease). We screened 153 cases with prenatally or clinically suspected X-chromosomal hydrocephalus for L1CAM mutations by SSCP analysis of the 28 coding exons and regulatory elements in the 5'-untranslated region of the gene. Forty-six pathogenic mutations were found (30.1% detection rate), the majority consisting of nonsense, frameshift, and splice site mutations. In eight cases, segregation analysis disclosed recent de novo mutations. Statistical analysis of the data indicates a significant effect on mutation detection rate of (i) family history, (ii) number of L1-disease typical clinical findings, and (iii) presence or absence of signs not typically associated with L1CAM-disease. Whereas mutation detection rate was 74.2% for patients with at least two additional cases in the family, only 16 mutations were found in the 102 cases with negative family history (15.7% detection rate). Our data suggest a higher than previously assumed contribution of L1CAM mutations in the pathogenesis of the heterogeneous group of congenital hydrocephalus. PMID- 10797422 TI - Rubinstein-Taybi syndrome caused by a De Novo reciprocal translocation t(2;16)(q36.3;p13.3). AB - Rubinstein-Taybi syndrome (RTS) is a multiple congenital anomalies and mental retardation syndrome characterized by facial abnormalities, broad thumbs, and broad big toes. We have shown previously that disruption of the human CREB binding protein (CBP) gene, either by gross chromosomal rearrangements or by point mutations, leads to RTS. Translocations and inversions involving chromosome band 16p13.3 form the minority of CBP mutations, whereas microdeletions occur more frequently (approximately 10%). Breakpoints of six translocations and inversions in RTS patients described thus far were found clustered in a 13-kb intronic region at the 5' end of the CBP gene and could theoretically only result in proteins containing the extreme N-terminal region of CBP. In contrast, in one patient with a translocation t(2;16)(q36.3;p13.3) we show by using fiber FISH and Southern blot analysis that the chromosome 16 breakpoint lies about 100 kb downstream of this breakpoint cluster. In this patient, Western blot analysis of extracts prepared from lymphoblasts showed both a normal and an abnormal shorter protein lacking the C-terminal domain, indicating expression of both the normal and the mutant allele. The results suggest that the loss of C-terminal domains of CBP is sufficient to cause RTS. Furthermore, these data indicate the potential utility of Western blot analysis as an inexpensive and fast approach for screening RTS mutations. PMID- 10797423 TI - X-linked congenital ataxia: a clinical and genetic study. AB - We report on a family in which two males are affected with X-linked congenital ataxia (XCA). Clinical manifestations include severe hypotonia at birth, delay of early motor development, slow eye movements, and nonprogressive cerebellar ataxia. The neurological examination excluded a neuromuscular disease, mental retardation, and pyramidal tract involvement. Neuroimaging showed global cerebellar atrophy in both patients that was not evident in the first years of life. The clinical findings in this family are very similar to those in a Russian pedigree [Illarioskin et al., 1996: Ann Neurol 40:75-83] and outline a recognizable phenotype. Linkage studies in our family, using 28 highly polymorphic Genethon microsatellite markers evenly distributed along the X chromosome, excluded a 24 cM interval between DXS990 and DXS424 located within the previous candidate region of 54 cM, reducing the critical interval. PMID- 10797424 TI - Genes on the X chromosome are important in undiagnosed mental retardation. AB - The clinical genetic diagnosis was reviewed in 429 subjects with intellectual disability in the Australian Child and Adolescent Development (ACAD) study of behavioural problems. With minor differences, the overall "general distribution by causation" was similar to that to that found by the Consensus Conference of the American College of Medical Genetics in 1995. There was a significant male excess in the whole series which was shown to reside in those with "autism," those with undiagnosed nonsyndromic mental retardation (NSMR) and those with X linked monogenic disorders. It is argued that a substantial proportion of undiagnosed NSMR is caused by genes on the X chromosome. Some of the practical problems of assigning individuals to diagnostic groups are discussed. PMID- 10797425 TI - OEIS complex--a population study. AB - Using a novel method for the analysis of infants with multiple malformations, we investigated the cluster of associated malformations called the OEIS (omphalocele, bladder exstrophy, imperforate anus, spine defect) complex among 5,260 infants with multiple malformations identified in four large registers of congenital malformations, corresponding to 5.84 million births. The existence of the OEIS complex was clearly demonstrated and malformations entering it could be defined. Other than the four classical malformations, omphalocele, bladder exstrophy, imperforate anus, and spine malformation, a strong association with spina bifida and intersex was stressed. Spine malformations occurred not only in the lumbosacral level but also more cranially, and an association also with upper spina bifida could be demonstrated. No specific association with any other malformation, including cardiac defects, was apparent. The OEIS complex is an unusually clearly defined entity among the various nonrandom associations which have been described. PMID- 10797427 TI - Nonsense mutation of the alpha-actinin-3 gene is not associated with dystrophinopathy. PMID- 10797426 TI - Mosaic tetrasomy 8q: inverted duplication of 8q23.3qter in an analphoid marker. AB - We observed an analphoid marker chromosome stable through cell division in a 16 year-old girl with developmental delay, short stature, limb contractures, and ovaries containing multiple cysts. She also developed myasthenia gravis at 15 years. The marker chromosome, present in 75% of metaphases (and in 90% of transformed lymphoblastoid cells), was C-band negative, and had no pan alpha satellite sequences detectable by fluorescence in situ hybridization (FISH). The 8q origin of the marker was determined by use of subtelomeric probes and was confirmed by chromosome 8 painting probes. The marker was shown to be an inversion duplication of 8q when subtelomeric, telomeric, and c-myc FISH probes hybridized to both ends of the marker. The karyotype was 47,XX,+inv dup(8)(qter- > q23.3::q23.3-->[neocen]-->qter), resulting in tetrasomy for 8q23.3qter. The parents had normal karyotypes. Centromeric proteins CENP-C and CENP-E were present, but alpha associated centromere protein CENP-B was absent at a position defining a neocentromere. PMID- 10797428 TI - Infant C677T mutation in MTHFR, maternal periconceptional vitamin use, and risk of nonsyndromic cleft lip. PMID- 10797429 TI - Inner ear abnormalities in Kabuki make-up syndrome: report of three cases. AB - Three patients, a female and two males, 28, 15, and 14 years of age, with Kabuki make-up syndrome (KMS) were studied for middle and inner ear abnormalities by using CT scanning of the petrous bones. All three patients had bilateral dysplasia of the inner ear, i.e., hypodysplasia of the cochlea, vestibule, and semicircular canals (so-called Mondini dysplasia), whereas their middle ears had no abnormalities. Audiometry demonstrated a sharp decrease in hearing of the high tone range, bilateral in one and unilateral in another, while the third patient was noncooperative. In view of these findings, it would be advisable to study each individual with KMS and hearing impairment for possible inner ear abnormalities. PMID- 10797430 TI - Heterogeneous mutations in the glucose-6-phosphatase gene in Japanese patients with glycogen storage disease type Ia. AB - Glycogen storage disease type Ia (GSD-Ia) is an autosomal recessive disorder of glycogen metabolism caused by glucose-6-phosphatase (G6Pase) deficiency. It is characterized by short stature, hepatomegaly, hypoglycemia, hyperuricemia, and lactic acidemia. Various mutations have been reported in the G6Pase gene (G6PC). However, in Japanese patients, a g727t substitution was found to be the major cause of GSD-Ia, accounting for 20 of 22 mutant alleles [Kajihara et al., 1995], and no other mutations have been found in this population. We analyzed four Japanese GSD-Ia patients and identified three other mutations in addition to the g727t. They included two missense mutations (R83H and P257L) and one nonsense mutation (R170X). Each of the three mutations exhibited markedly decreased G6Pase activity when expressed in COS7 cells. A patient homozygous for R170X showed multiple episodes of profound hypoglycemia associated with convulsions, while P257L was associated with a mild clinical phenotype. The presence of R170X in three unrelated families may implicate that it is another important mutation in the etiology of GSD-Ia in Japanese patients. Thus, the detection of non-g727t mutations is also important in establishing the DNA-based diagnosis of GSD-Ia in this population. PMID- 10797431 TI - Widely distributed mutations in the COL2A1 gene produce achondrogenesis type II/hypochondrogenesis. AB - The COL2A1 gene was assayed for mutations in genomic DNA from 12 patients with achondrogenesis type II/hypochondrogenesis. The exons and flanking sequences of the 54 exons in the COL2A1 gene were amplified by a series of specific primers using PCR. The PCR products were scanned for mutations by conformation sensitive gel electrophoresis, and PCR products that generated heteroduplex bands were then sequenced. Mutations in the COL2A1 gene were found in all 12 patients. Ten of the mutations were single base substitutions that converted a codon for an obligate glycine to a codon for an amino acid with a bulkier side chain. One of the mutations was a change in a consensus RNA splice site. Another was an 18-base pair deletion of coding sequences. The results confirmed previous indications that conformation sensitive gel electrophoresis is highly sensitive for detection of mutations in large and complex genes. They also demonstrate that most, if not all, patients with achondrogenesis type II/hypochondrogenesis have mutations in the COL2A1 gene. PMID- 10797432 TI - Mosaic rearrangement of chromosome 18: characterization by FISH mapping and DNA studies shows trisomy 18p and monosomy 18p both of paternal origin. AB - Structural abnormalities of chromosome 18p mainly consist of isochromosomes of the short arm, which result in tetrasomy 18p. Trisomy 18p is much rarer, and less well characterized. We report on a 12-year-old girl with minor facial anomalies, delayed development, abnormal hands, atopic dermatitis, and hearing loss. She was mosaic for two abnormal cell lines in peripheral blood. In 90% of cells, a dicentric chromosome with duplication of the whole short arm of chromosome 18 resulted in trisomy 18p; 10% of cells had monosomy 18p, arising from a t(14;18)(p11;q11). FISH mapping, with multiple region specific and locus specific probes from the short and long arm of chromosome 18, showed that the structure of the dicentric chromosome 18 was 18pter-->18q23::18q11-->18pter. DNA polymorphisms for chromosome 18 showed that the abnormalities of chromosome 18 were paternal in origin. Combining all results, we could link the trisomy 18p and monosomy 18p to a common origin via a complex series of events in an early mitosis. PMID- 10797433 TI - Embryonal rhabdomyosarcoma and chromosomal breakage in a newborn infant with possible Dubowitz syndrome. AB - We report on a newborn girl with Dubowitz syndrome (DS) and embryonal rhabdomyosarcoma (ERMS), with multiple chromosomal breakage (MCB). The tumor was resected but recurred in a few months, resulting in the infant's death. Malignancy and chromosomal breakage have been reported previously in DS. However, ERMS has not been reported among the malignant tumors diagnosed in DS. To our knowledge, concurrence of DS, ERMS, and MCB has not been reported previously. This is the first observation of DS in the Arab ethnic group. PMID- 10797434 TI - Proportion of cells with paternal 11p15 uniparental disomy correlates with organ enlargement in Wiedemann-beckwith syndrome. AB - "Genetic mosaicism" describes the presence of two or more populations of cells within a single individual that differ in their genomic constitution. Although the occurrence of asymmetric overgrowth in Wiedemann-Beckwith syndrome (WBS) suggests that mosaicism has some role in the WBS phenotype, no direct evidence for this has been published. WBS is a congenital overgrowth syndrome with variable phenotype linked to the imprinted gene cluster on chromosome region 11p15. We have performed a molecular survey of multiple organs and tissues in a case of WBS with a high degree of mosaic paternal 11p15 uniparental disomy (UPD). The organs most severely affected were those with the highest percentage of cells with UPD. In particular there was a striking difference in the degree of mosaicism for 11p15 UPD between the extremely enlarged left adrenal and non enlarged right adrenal gland. This result indicates that the proportion of paternal 11p15 UPD cells correlates with the tissue phenotype of WBS. Our results suggest that high proportions of abnormal cells result from a combination of stochastic events and cell selection. Mosaicism may explain the variable phenotypes including hemihyperplasia and predisposition to childhood cancers in WBS patients. PMID- 10797435 TI - Brown-Vialetto-Van Laere syndrome in a large inbred Lebanese family: confirmation of autosomal recessive inheritance? AB - Brown-Vialetto-Van Laere syndrome or pontobulbar palsy with deafness is a rare disorder characterized by bilateral nerve deafness, a variety of cranial nerve disorders usually involving the motor components of the 7th and 9th to 12th cranial nerves, and less commonly an involvement of spinal motor nerves and upper motor neurons. Familial and sporadic cases have been reported. Based on particular evidence, autosomal recessive, autosomal dominant, and X-linked inheritance, as well as autoimmune origin have been considered. We report on a large inbred Lebanese family with four patients of both sexes, strongly suggesting autosomal recessive inheritance. PMID- 10797436 TI - Brazilian family with pure autosomal dominant spastic paraplegia maps to 8q: analysis of muscle beta 1 syntrophin. AB - The autosomal dominant hereditary spastic paraplegias (AD-HSP) are a heterogeneous group of degenerative disorders of the central motor system, characterized by progressive spasticity of the lower limbs. Five loci for pure AD HSP have been identified to date: SPG3 at 14q, SPG4 at 2p, SPG6 at 15q, SPG8 at 8q, and more recently SPG10 at 12q. We have analyzed a Brazilian family with 16 affected individuals by pure AD-HSP who developed progressive gait disturbance with onset at age 18-26 years. Linkage analysis performed with 13 relatives (6 affected and 7 normal) excluded SPG3, SPG4, and SPG6 as candidate regions. However, positive LOD scores were obtained with markers flanking the candidate region for the SPG8 locus [maximum two point Lod score (Zmax) = 3.3 at theta = 0 for D8S1804]. In this region lies the syntrophin beta 1 gene (SNT2B1), a widely expressed dystrophin-associated protein and therefore a good positional and functional candidate for this disease. Immunohistochemical and Western Blot (WB) studies showed that the distribution, expression, and apparent molecular weight of the beta 1 syntrophin protein were comparable to those of normal control individuals. Therefore, it is unlikely that defects in this protein are related to SPG8, at least in the present family. PMID- 10797437 TI - Metacarpophalangeal pattern profile analysis in Noonan syndrome. AB - Metacarpophalangeal pattern (MCPP) analysis is an application of an anthropometric technique that provides a quantitative assessment of the amount and direction of abnormality in the hand skeleton. MCPP analysis was undertaken on 15 individuals (9 males, 6 females) with Noonan syndrome ranging in age from 0.1 to 36 years with a mean age at 11.6 years. The overall average Z score for the MCPP variables was -2.1 and the range was -2.5 (for metacarpal two) and -1.5 (for middle phalanx 5). The average hand pattern variability index, a measure of hand bone length relationships, was abnormal. A Pearsonian correlation analysis was used to assess similarity between the mean pattern and each of the 15 individual patterns. Nine (60%) of the fifteen individuals with Noonan syndrome had significant positive correlations (P < 0.05), indicating homogeneity or similarity in the hand patterns. A stepwise discriminant analysis was performed on Z score data from the individual hand bone measurements on the 15 subjects with Noonan syndrome and 41 healthy controls (24 females, 17 males; mean age = 13.1 years with age range of 9.6 to 18 years). This analysis produced a discriminant function with two MCPP variables (metacarpal 1 and middle phalanx 3) entering into the function and producing a correct classification rate of 93%. The two MCPP variables contributed to the overall difference between individuals with Noonan syndrome and the normative sample. The hand pattern variability index was outside of the normal range, indicating an abnormal MCPP with multivariate analysis. The MCPP analysis may be useful as a tool for diagnosis in screening subjects for Noonan syndrome. PMID- 10797438 TI - Clinic-based study of plexiform neurofibromas in neurofibromatosis 1. AB - Individuals with neurofibromatosis 1 (NF1) develop both benign and malignant tumors at an increased frequency. One of the most common benign tumors in NF1 is the plexiform neurofibroma. These tumors cause significant morbidity and mortality on account of their propensity to grow and affect adjacent normal tissues. To determine the clinical profile of plexiform neurofibromas in NF1, we conducted a retrospective review of 68 NF1 patients with plexiform neurofibroma. In our series, 44% of tumors were detected by 5 years of age and most were located in the trunk and extremities. Only two patients developed malignant peripheral nerve sheath tumors in their preexisting plexiform neurofibromas. Lastly, we demonstrate that there were no specific clinical features of NF1 associated with the presence of plexiform neurofibroma. These results underscore the importance of careful serial examinations in the evaluation of patients with NF1. PMID- 10797439 TI - Investigation of germline PTEN, p53, p16(INK4A)/p14(ARF), and CDK4 alterations in familial glioma. AB - Epidemiological studies suggest that some familial aggregations of glioma may be due to inherited predisposition. Many genes involved in familial cancers are frequently altered in the corresponding sporadic forms. We have investigated several genes known to be altered in sporadic gliomas for their potential contribution to familial glioma. Fifteen glioma patients with a family history of brain tumors were identified through the Mayo Clinic Department of Neurology (nine diffuse astrocytomas, two oligodendrogliomas, two mixed oligoastrocytomas, one pilocytic astrocytoma, and one pineal glioma). Eleven of the propositi had one or more first degree relative with a glioma. Lymphocyte DNA was derived from each of the patients and analyzed by polymerase chain reaction (PCR) and direct sequencing of the PTEN, p53, p16(INK4A)/p14(ARF), and CDK4 genes. In addition, fluorescence in situ hybridization (FISH) was performed on EBV-transformed lymphocytes from each affected individual to detect germline copy number of the p16(INK4A)/p14(ARF) tumor suppressor region. A p53 germline point mutation was identified in one family with some findings of Li-Fraumeni syndrome, and a hemizygous germline deletion of the p16(INK4A)/p14(ARF) tumor suppressor region was demonstrated by FISH in a family with history of both astrocytoma and melanoma. Thus, whereas germ-line mutations of PTEN, p53, p16(INK4A)/p14(ARF), and CDK4 are not common events in familial glioma, outside of familial cancer syndromes, point mutations of p53 and hemizygous deletions and other rearrangements of the p16(INK4A)/p14(ARF) tumor suppressor region may account for a subset of familial glioma cases. Collectively, these data lend genetic support to the heritable nature of some cases of glioma. PMID- 10797440 TI - X-linkage does not account for the absence of father-son similarity in plasma uric acid concentrations. AB - Plasma uric acid concentration aggregates in families, and this similarity has been suggested to be due, in part, to multiple shared genes. Men have higher plasma uric acid concentrations than women and are affected with gout nine times more frequently. Rare forms of hyperuricemia and gout are due to mutations of X linked genes (HPRT1 and PRPS1). Given these observations, we tested the hypothesis that normal variation in plasma uric acid levels would display a pattern of familial similarity consistent with X-linkage in 892 individuals from 196 obese but otherwise healthy families. As predicted by X-linked inheritance, fathers and sons showed no resemblance in plasma uric acid concentration (r = 0.013, NS), while all other pairings showed moderate-to-strong familial resemblance (ranging from 0.167, P < 0.01, parent-offspring to 0.415, sister sister, P < 0.01). We then tested the hypothesis that loci along the X chromosome would influence plasma uric acid concentration. We conducted both single-point and multipoint linkage analyses using 17 X-linked markers spaced at approximately 9 cm intervals to determine whether allele sharing among sibs was related to sib similarity in plasma uric acid concentrations (n = 1,100 sib pairs). We found no regions of the X chromosome that cosegregated with plasma uric acid concentrations (P > 0.05). We conclude that variation in genes on the X chromosome contribute little to normal variation in plasma uric acid concentrations. PMID- 10797441 TI - Molecular cytogenetic characterization of a small, familial supernumerary ring chromosome 7 associated with mental retardation and an abnormal phenotype. AB - A family is described in which a mother and two of her children were mosaic for a small supernumerary ring chromosome. As the origin of the ring chromosome could not be determined by routine cytogenetic studies, fluorescent in situ hybridization was performed, which indicated that the ring chromosome was derived from the pericentromeric region of chromosome 7. Further characterization with a YAC-probe showed the involvement of the proximal q-arm of chromosome 7. Both sibs had speech difficulties and were mildly mentally retarded whereas the mother's intelligence was at the lower end of the normal range. They all had an unusual face, characterized by a flat profile, short forehead, downslant of the palpebral fissures, high and broad nasal bridge, simply formed ears, and prognathia. This is the second report of a small supernumerary ring chromosome derived from the pericentromeric region of chromosome 7, and the described clinical phenotype differs from that delineated in the previous report. PMID- 10797442 TI - Rare case of De Novo interstitial deletion 2q13q21: clinical, cytogenetic, and molecular studies. PMID- 10797443 TI - Narrowing the map of a gene (MRXS9) for X-linked mental retardation, microcephaly, and variably short stature at Xq12-q21.31. PMID- 10797444 TI - Craniosynostosis: molecular testing-a necessity for counseling. PMID- 10797445 TI - Three-color versus four-color multiparameter cell cycle analyses of primary acute myeloid leukemia samples. AB - Checkpoint alterations that impact cell cycle and apoptosis responses to therapeutic treatments may produce drug resistance in acute myeloid leukemia (AML). To study these, we have developed flow cytometry assays of checkpoint function that also allow quantitation of key molecular regulators of apoptosis and cell cycle. We have used three-color (3C) assays, with FITC-labeled anti-BCL 2 and PE-labeled anti-proliferating cell nuclear antigen (PCNA) antibodies, and the DNA dye 7-aminoactinomycin, to characterize primary leukemia cells identified in DNA x side light scatter (SSC) histograms. We showed that 3C assays are accurate and reproducible in analyses of leukemia cell lines and of primary AML and normal bone marrow samples (Banker et al.: Blood 89: 243-255, 1997; Banker et al.: Leukemia Res 22: 221-239, 1998; Banker et al.: Clin Cancer Res 4: 3051-3062, 1998). To further confirm the validity of our SSC leukemia cell gating and to address whether immunophenotypic AML subsets might have different biologic properties, we have now designed four-color (4C) flow assays to characterize checkpoint status in leukemic blasts specifically identified by surface immunostaining. In modeling this assay strategy, PE/Cy5-labeled anti-CD34 antibody was used to detect blasts, with FITC-labeled anti-BCL-2, PE-labeled anti PCNA antibodies, and Hoechst 33342 (H33342) DNA dye. Four-color CD34-gated data was concordant with 3C, SSC-gated data for leukemia cell lines and for most primary AML samples with high and intermediate blast counts. BCL-2 and PCNA immunopositivity and sub-G1 apoptosis determinations were different in the CD34 gated versus SSC-gated blasts in particular samples with smaller CD34(+) subsets, suggesting that leukemia samples can contain blast subsets with different biologic properties. On the other hand, PCNA-gated cell-cycle distributions in untreated cells and G1 versus S phase cell-cycle arrests after cytosine arabinoside treatments were completely concordant in 4C and 3C assays. We conclude that both 3C and 4C assays can be used to characterize protein expression and cell-cycle drug response patterns in leukemia blasts, but that 4C assays may additionally allow discrimination of these properties in immunophenotypic leukemia subsets. PMID- 10797446 TI - External quality assessment of flow cytometric HLA-B27 typing. AB - A biannual external quality assurance (EQA) scheme for flow cytometric typing of the HLA-B27 antigen is operational in The Netherlands and Belgium since 1995. We report here on the results of the first seven send-outs to which 36 to 47 laboratories participated. With the send-out, four specimens from blood bank donors, who had been typed for HLA Class I antigens by complement-dependent cytotoxicity, were distributed. Subtyping of the HLA-B27 allele was performed by PCR-SSP. Ten samples were HLA-B27(pos) (all HLA-B*2705) and 18 were HLA-B27(neg). For flow cytometry, the most widely monoclonal antibody (MoAb) used was FD705, followed by GS145.2 and ABC-m3. The majority of laboratories used more than 1 anti-HLA-B27 MoAb for typing. The HLA-B27(pos) samples were correctly classified as positive by the large majority of participants (median 95%; range 85% to 100% per send out); some participants considered further typing necessary and misclassification as negative was only sporadically seen. The classification of HLA-B27(neg) samples as negative was less straightforward. Ten samples were correctly classified as such by 97% (82% to 100%) of the participants, whereas 64% (range 53% to 70%) of the participants classified the remaining eight samples as HLA-B27(neg). There was no significant prevalence of a particular HLA-B allele among these eight "poor concordancy" samples as compared to the ten "good concordancy" samples. Inspection of the reactivity patterns of the individual MoAb with HLA-B27(neg) samples revealed that ABC-m3 showed very little cross reactivity apart from its well-known cross-reactivity with HLA-B7, whereas the cross-reactivity patterns of GS145.2 and FD705 were more extensive. The small sample size (n = 18) and the distribution of HLA-B alleles other than HLA-B27 did not allow assignment of specificities to these cross-reactions. Finally, we showed that standardized interpretation of the combined results of two anti-HLA B27 MoAb reduced the frequency of false-positive conclusions on HLA-B27(neg) samples. In this series, the lowest frequency of false-positive assignments was observed with the combination of the FD705 and ABC-m3 MoAb. PMID- 10797447 TI - The "typical" immunophenotype of acute promyelocytic leukemia (APL-M3): does it prove true for the M3-variant? AB - The immunophenotypes of 12 acute promyelocytic leukemias (APL-M3; eight hypergranular, four microgranular) with documented PML-RAR-alpha fusion gene are presented. Bone marrow mononuclear cells were immunophenotyped using a panel of 20 monoclonal antibodies. The hypergranular APLs exhibited a mature myeloid phenotype as it has been described to be typical for M3. No lineage infidelity was detectable in classic M3 cases. In contrast, among the four cases of M3 variant, all leukemias showed marked expression of CD34 and two of four cases expressed the HLA-DR antigen. The CD2 antigen was expressed in three of four cases. Furthermore, one case showed expression of the CD56 antigen, and one case was positive for the blood group H antigen. The data suggest that microgranular APL is a heterogeneous entity with regard to the immunologic phenotype. PMID- 10797448 TI - Automated analysis in flow cytometry. PMID- 10797449 TI - Diagnosis of unexpected acute myeloid leukemia and chronic lymphocytic leukemia: a case report demonstrating the perils of restricted panels in flow cytometric immunophenotyping. AB - We report on the flow cytometric identification of concomitant acute myeloid leukemia and chronic lymphocytic leukemia in cytology specimens submitted with minimal clinical information. A 64-year-old man presented with fever and progressive dyspnea on exertion. Chest X-ray and computed tomography scan showed a left upper lobe pulmonary mass. Pulmonary capillary pullback specimens were collected to determine infectious verses neoplastic etiology. The pulmonary capillary pullback specimens showed atypical mononuclear cells with enlarged, slightly irregular nuclei; visible nucleoli; and basophilic cytoplasm. Flow cytometric analysis of the specimen for lymphoma was requested. Flow cytometric immunophenotypic studies showed that 78% of the cells were CD34 positive, CD45 dim positive and CD11c positive, consistent with acute myeloid leukemia. About 0. 75% of the cells expressed CD5 as well as dim CD20 and were monoclonal for kappa light chains: consistent with chronic lymphocytic leukemia/small lymphocytic lymphoma. At this time the clinician communicated a history of myelodysplastic syndrome of refractory anemia subtype. Peripheral blood was obtained for further immunophenotyping and the patient was immediately treated for his acute myeloid leukemia. This case demonstrates that a diagnostic antibody panel should allow evaluation of all cell types as per the U.S./Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry (Stewart et al.: Cytometry 30:231-235, 1997). Published 2000 Wiley-Liss, Inc. PMID- 10797450 TI - Immunophenotype of a transient myeloproliferative disorder in a newborn with trisomy 21. AB - Cytologic, immunologic, and cytogenetic studies were performed on the blast cells of a newborn with Down syndrome and transient myeloproliferative disease. This hematologic disorder is uncommon, and occurs primarily in infants with Down syndrome. This boy presented with a high white blood cell count and a high percentage of blast cells, without anemia or thrombocytopenia. Chromosome analysis showed a constitutional trisomy 21 without any other clonal abnormality. A three-color flow cytometric analysis was performed and revealed two different CD45 dim, CD34(+), CD117(+), CD56(+) immature subpopulations: the normal immature myeloid precursor and an immature blast cell population that expressed CD41, CD42, CD61, CD36, CD13, CD1a, and CD2. We postulate that this population could be the leukemic precursor involved in the acute megakaryoblastic leukemia frequently observed in children with Down syndrome. PMID- 10797451 TI - Laser scanning cytometry can complement the flow cytometric DNA analysis in paraffin-embedded cancer samples: a paradigmatic case. AB - Archival studies on paraffin-embedded tumor samples are often complicated by difficulty obtaining a reliable diploid DNA standard. Nontumor cells, e.g., inflammatory and stromal cells, most often found interspersed among tumor cells, would represent a solution to this problem. Unfortunately, there is an inherent difficulty to positively identifying tumor cells in paraffin-embedded specimens. Using an aneuploid paraffin-embedded breast cancer sample, we show here that laser scanning cytometer (LSC) in conjunction with flow cytometry can help to address this issue. Following standard protocols, the tissue was deparaffinized and rehydrated, and the nuclei mechanically isolated before being exposed to propidium iodide. An aliquot served for single-parameter flow cytometric analysis, and the remaining cells were cytocentrifuged onto a microscope slide and LSC analysis was performed. The DNA histogram profiles generated by the two approaches were comparable and both showed the presence of cell populations with different DNA content. To assess the nature of these subsets, we performed a correlated measurement of DNA content and chromatin organization at the single cell level by LSC. This allowed the identification of several subsets of nuclei. Slides were then stained with Giemsa and the nature of these subsets was assessed morphologically by exploiting the relocating capability of LSC. Inflammatory and stromal cells, residual diploid epithelial cells, and hyperdiploid tumor cells each characterized by a peculiar coordinate pattern of DNA content and chromatin organization-could be positively identified. Diploid, nontumor cells can then be used as an internal standard for DNA ploidy. PMID- 10797452 TI - Forum: journal club PMID- 10797453 TI - Comparative effects of extremely high power microwave pulses and a brief CW irradiation on pacemaker function in isolated frog heart slices. AB - The existence of specific bioeffects due to high peak power microwaves and their potential health hazards are among the most debated but least explored problems in microwave biology. The present study attempted to reveal such effects by comparing the bioeffects of short trains of extremely high power microwave pulses (EHPP, 1 micros width, 250-350 kW/g, 9.2 GHz) with those of relatively low power pulses (LPP, 0.5-10 s width, 3-30 W/g, 9.2 GHz). EHPP train duration and average power were made equal to those of an LPP; therefore both exposure modalities produced the same temperature rise. Bioeffects were studied in isolated, spontaneously beating slices of the frog heart. In most cases, a single EHPP train or LPP immediately decreased the inter-beat interval (IBI). The effect was proportional to microwave heating, fully reversible, and easily reproducible. The magnitude and time course of EHPP- and LPP-induced changes always were the same. No delayed or irreversible effects of irradiation were observed. The same effect could be repeated in a single preparation numerous times with no signs of adaptation, sensitization, lasting functional alteration, or damage. A qualitatively different effect, namely, a temporary arrest of preparation beats, could be observed when microwave heating exceeded physiologically tolerable limits. This effect also did not depend on whether the critical temperature rise was produced by LPP or EHPP exposure. Within the studied limits, we found no indications of EHPP-specific bioeffects. EHPP- and LPP-induced changes in the pacemaker rhythm of isolated frog heart preparation were identical and could be entirely attributed to microwave heating. PMID- 10797454 TI - A new in vitro exposure device for the mobile frequency of 900 MHz. AB - A wire patch cell has been designed for exposing cell cultures during in vitro experiments studying possible effects of mobile radio telephone. It is based on the wire patch antenna which works at 900 MHz with a highly homogeneous field inside the antenna cavity. The designed cell structure is symmetric and provides a rather homogeneous field distribution in a large area around its centre. Moreover, the exposure cell can irradiate equally up to eight 35 mm Petri dishes at the same time, which enhances the statistical biological studies. To improve the specific absorption rate (SAR) homogeneity inside each sample, each dish is placed into another 50 mm dish. This way, SAR inhomogeneity is always proper for biological studies (below 30%). The main advantage of this new device is that it can provide SAR levels 20 times higher than those induced by classical Crawford transverse electromagnetic (TEM) cell. Moreover, this small open device is easy to construct and fits into an incubator. However, to be used for in vitro, the wire patch cell is a radiating element with the same radiating pattern as a dipole, and thus some absorbing materials are necessary around the system when used for in vitro experiments. Secondly, because of its narrow bandwidth, it is difficult to maintain its working frequency. To overcome this problem, a matching device is integrated into the test cell. In this paper, we present a detailed explanation of the cell behavior and dosimetric assessments for eight 35 mm Petri dishes exposed. Simulations using the Finite Difference Time Domain technique and experimental investigations have been carried out to design the cell at 900 MHz. The numerical dosimetry was validated by dosimetric measurements. These investigations estimated the dosimetric precision at 11%. PMID- 10797455 TI - Reflection and absorption of millimeter waves by thin absorbing films. AB - Reflection, transmission, and absorption of mm-waves by thin absorbing films were determined at two therapeutic frequencies: 42. 25 and 53.57 GHz. Thin filter strips saturated with distilled water or an alcohol-water solution were used as absorbing samples of different thicknesses. The dependence of the power reflection coefficient R(d) on film thickness (d) was not monotonic. R(d) passed through a pronounced maximum before reaching its steady-state level [R(infinity)]. Similarly, absorption, A(d), passed two maximums with one minimum between them, before reaching its steady-state level [A(infinity)]. At 42.25 GHz, A(d) was compared with absorption in a semi-infinite water medium at a depth d. When d < 0.3 mm, absorption by the film increased: at d = 0.1 mm the absorption ratio for the thin layer sample and the semi-infinite medium was 3.2, while at d = 0.05 mm it increased up to 5.8. Calculations based on Fresnel equations for flat thin layers adequately described the dependence of the reflection, transmission, and absorption on d and allowed the determination of the refractive index (n), dielectric constant (epsilon), and penetration depth (delta) of the absorbing medium for various frequencies. For water samples, epsilon was found to be 12.4-19.3j, delta = 0.49 mm at 42.25 GHz, and epsilon = 9.0-19.5j, delta = 0.36 mm at 53.57 GHz. The calculated power density distribution within the film was strongly dependent on d. The measurements and calculations have shown that the reflection and absorption of mm-waves by thin absorbing layers can significantly differ from the reflection and absorption in similar semi-infinite media. The difference in reflection, absorption, and power density distribution in films, as compared to semi-infinite media, are caused by multiple internal reflections from the film boundaries. That is why, when using thin phantoms and thin biological samples, the specifics of the interaction of mm-waves with thin films should be taken into account. PMID- 10797456 TI - Directed and enhanced neurite growth with pulsed magnetic field stimulation. AB - Pulsed magnetic field (PMF) stimulation was applied to mammalian neurons in vitro to influence axonal growth and to determine whether induced current would direct and enhance neurite growth in the direction of the current. Two coils were constructed from individual sheets of copper folded into a square coil. Each coil was placed in a separate water-jacketed incubator. One was energized by a waveform generator driving a power amplifier, the other was not energized. Whole dorsal root ganglia (DRG) explant cultures from 15-day Sprague-Dawley rat embryos were established in supplemented media plus nerve growth factor (NGF) at concentrations of 0-100 ng/mL on a collagen-laminin substrate. Dishes were placed at the center of the top and bottom of both coils, so that the DRG were adjacent to the current flowing in the coil. After an initial 12 h allowing DRG attachment to the substrate floor, one coil was energized for 18 h, followed by a postexposure period of 18 h. Total incubation time was 48 h for all DRG cultures. At termination, DRG were histochemically stained for visualization and quantitative analysis of neurite outgrowth. Direction and length of neurite outgrowth were recorded with respect to direction of the current. PMF exposed DRG exhibited asymmetrical growth parallel to the current direction with concomitant enhancement of neurite length. DRG cultures not PMF exposed had a characteristic radial pattern of neurite outgrowth. These results suggest that PMF may offer a noninvasive mechanism to direct and promote nerve regeneration. PMID- 10797457 TI - Extremely low frequency magnetic fields can either increase or decrease analgaesia in the land snail depending on field and light conditions. AB - Results of prior investigations with opioid peptide mediated antinociception or analgaesia have suggested that these extremely low frequency (ELF) magnetic field effects are described by a resonance mechanism rather than mechanisms based on either induced currents or magnetite. Here we show that ELF magnetic fields (141 414 microT peak) can, in a manner consistent with the predictions of Lednev's parametric resonance model (PRM) for the calcium ion, either (i) reduce, (ii) have no effect on, or (iii) increase endogenous opioid mediated analgaesia in the land snail, Cepaea nemoralis. When the magnetic fields were set to parameters for the predictions of the PRM for the potassium ion, opioid-peptide mediated analgaesia increased and there was evidence of antagonism by the K(+) channel blocker, glibenclamide. Furthermore, these effects were dependent on the presence of light; the effects were absent in the absence of light. These observed increases and decreases in opioid analgaesia are largely consistent with the predictions of Lednev's PRM. PMID- 10797458 TI - Effects of 50 Hz electric currents and magnetic fields on the prokaryote Propionibacterium acnes. AB - The effects of 50 Hz sinusoidal electric currents and magnetic fields on the Gram positive skin bacterium Propionibacterium acnes were investigated. Intracellular free calcium ([Ca(2+)](i)), intracellular pH (pH(i)), and cell viability were examined, based on their relevance to ELF field studies and on previous studies conducted on P. acnes (UVA irradiation, photosensitization using porphyrin-based sensitizers, and broad-band red light). The [Ca(2+)](i) and the pH(i) were measured spectrofluorimetrically using the fluorescent probes fura-2 and BCECF, respectively. Sham-exposed controls were used to assess the field exposed samples. Cell suspensions were exposed to 50 Hz, 0.2 mT sinusoidal magnetic fields generated by using Helmholtz coils for up to 30 min. The estimated maximum induced electric field was 0.2 mV/m. Changes in [Ca(2+)](i) and cell viability were not detected. Ag/AgCl electrodes were used to expose cell suspensions to 50 Hz sinusoidal electric currents. The current densities were in the range 0.015 1500 A/m(2) (corresponding electric fields congruent with0.01-1000 V/m). Changes in [Ca(2+)](i) were not observed after current exposure. Current densities of 800 A/m(2) (electric field E congruent with550 V/m) were required for a 50% reduction in cell viability. Current densities greater than 800 A/m(2) were required for a reduction in pH(i). However, a pH gradient across the cell membrane (inside alkaline) was maintained even when exposure resulted in less than 0. 2% survival (1400 A/m(2), E congruent with950 V/m). Thus, dissipation of the pH gradient across the cell membrane and changes in [Ca(2+)](i) were not a consequence of cell inactivation by 50 Hz electric currents. This is in contrast to inactivation of P. acnes by UVA irradiation or photosensitization, where such changes have been obtained. PMID- 10797459 TI - Zeeman-Stark modeling of the RF EMF interaction with ligand binding. AB - The influence of radiofrequency electromagnetic exposure on ligand binding to hydrophobic receptor proteins is a plausible early event of the interaction mechanism. A comprehensive quantum Zeeman-Stark model has been developed which takes into account the energy losses of the ligand ion due to its collisions inside the receptor crevice, the attracting nonlinear endogenous force due to the potential energy of the ion in the binding site, the out of equilibrium state of the ligand-receptor system due to the basal cell metabolism, and the thermal noise. The biophysical "output" is the change of the ligand binding probability that, in some instances, may be affected by a suitable low intensity exogenous electromagnetic "input" exposure, e.g., if the depth of the potential energy well of a putative receptor protein matches the energy of the radiofrequency photon. These results point toward both the possibility of the electromagnetic control of biochemical processes and the need for a new database of safety standards. PMID- 10797460 TI - Comment: "Microwave irradiation influences on the state of human cell nuclei" [Y.G. Shckorbatov Et al., 19:414-419 (1998)]. PMID- 10797461 TI - Reply: some considerations about the method of intracellular microelectrophoresis. PMID- 10797462 TI - The impact of Genomics' technologies on pharmaceutical research. PMID- 10797463 TI - The Human Genome Project--an overview. AB - The human genome sequence will underpin human biology and medicine in the next century, providing a single, essential reference to all genetic information. The international program to determine the complete DNA sequence (3,000 million bases) is well underway. As of January 2000, 50% of the sequence is available in the public domain. A comprehensive working draft is expected this year, and the entire sequence is projected to be finished in 2003. DNA sequencing is carried out on mapped, overlapping bacterial clones of 150-200 kb. The working draft comprises assembled unfinished sequence and is released immediately in the public domain. The draft sequence of each clone is then completed, by closing any remaining gaps and resolving any ambiguities, before the entire sequence is checked, annotated, and submitted to the public databases. The sequence of each clone is finished to an accuracy of >99.99%. The availability of a reference sequence of the genome provides the basis for studying the nature of sequence variation, particularly single nucleotide polymorphisms (SNPs), in human populations. SNP typing is a powerful tool for genetic analysis, and will enable us to uncover the association of loci at specific sites in the genome with many disease traits. SNPs occur at a frequency of approximately 1 SNP/kb throughout the genome when the sequence of any two individuals is compared. Programs to detect and map SNPs in the human genome are underway with the aim of establishing a SNP map of the genome during the next two years. The human genome sequence will provide a complete description of all the genes. Annotation of the sequence with the gene structures is achieved by a combination of computational analysis (predictive and homology-based) and experimental confirmation by cDNA sequencing. Detecting homologies between newly defined gene products and proteins of known function helps to postulate biochemical functions for them, which can then be tested. Establishing the association of specific genes with disease phenotypes by mutation screening, particularly for monogenic disorders, provides further assistance in defining the functions of some gene products, as well as helping to establish the cause of the disease. As our knowledge of gene sequences and sequence variation in populations increases, we will pinpoint more and more of the genes and proteins that are important in common, complex diseases. A more detailed understanding of the function of the human genome will be achieved as we identify sequences that control gene expression. Given the availability of gene sequences, the expression status of genes in particular tissues can be monitored in parallel. By comparing corresponding genomic sequences in different species (for example: man, mouse, chicken, and zebrafish), regions that have been highly conserved during evolution can be identified, many of which reflect conserved functions such as gene regulation. These approaches promise to greatly accelerate our interpretation of the human genome sequence. PMID- 10797464 TI - High throughput gene expression screening: its emerging role in drug discovery. AB - The genetic makeup and the environment influences the health and welfare of an individual. At both the tissue and cellular level, physiological function can be correlated with the transcription of genes, whose protein products contribute and influence the activity of biological systems. In order to understand these processes, it is therefore essential to determine the temporal and spatial patterns of gene expression, and, with particular relevance to drug discovery, define changes that occur during development of disease or treatment with therapeutic agents. PMID- 10797465 TI - Genetics, genomics, and drug discovery. AB - Genetics and genomics are altering the drug discovery process and changing the practice of medicine. Genetic mapping and positional cloning will lead to the identification of disease susceptibility alleles of different genes and this will be further enabled by the publication of the DNA sequence of the human genome. Single nucleotide polymorphisms discovered by comparative sequencing and other methods will also speed up positional cloning and allow pharmacogenetics to proceed in a more comprehensive way than heretofore. Gene expression arrays coupled with studies in model organisms will help to define the function of all genes and determine how the products interact-the next big step. High throughput protein structure determination will also provide large-scale descriptions of protein function but with more biochemical clarity. In all of these studies data integration remains the biggest challenge. PMID- 10797466 TI - PROfusion: genetically tagged proteins for functional proteomics and beyond. AB - As a new enabling technology in functional genomics, Phylos, Inc. has pioneered PROfusiontrade mark technology, whereby proteins are covalently tagged with their own genetic information (mRNA). Using this technology, both synthetic and natural libraries (representing the repertoire of proteins naturally expressed in a given cell type or tissue source) have been constructed. Due to the in vitro nature of library construction, Phylos libraries are the largest described to date, up to 10(14) in size. From a library of such molecules, one can select for a protein function of choice with the benefit that the genetic material is linked to the protein for subsequent PCR amplification, enrichment, and, ultimately, identification. PMID- 10797467 TI - Mouse genetics/genomics: an effective approach for drug target discovery and validation. AB - The mouse has become the premier mammalian system for the identification of the genetic basis of both mono- and oligogenic disorders, as well as the understanding of complex diseases with gene-gene and gene-environment interactions. The similarity between human and mouse genetic disease is sometimes striking, while in other cases the phenotypes are less similar. The ability to genetically map and then clone single gene disorders rapidly, and the emerging technologies that will allow the economical identification of the polygenes controlling quantitative traits further demonstrate the utility of the mouse as a model for gene discovery. Additionally, the ability to genetically manipulate the mouse through transgenesis and gene targeting allows for the testing of hypotheses regarding specific gene function and their role in disease. The utility of the mouse extends beyond being just a gene discovery tool to provide prevalidated targets. It can also be used for the development of animal models, and the testing of compounds in specifically constructed transgenic and knockout strains to further define the target and pathway of a therapeutic compound. PMID- 10797468 TI - DNA adducts produced by oils, oil fractions and polycyclic aromatic hydrocarbons in relation to repair processes and skin carcinogenesis. AB - Ten polycyclic aromatic hydrocarbons (PAHs) mainly with three or four aromatic rings were tested for their ability to induce DNA adduct formation in mouse skin. Four of these were selected to investigate adduct formation and loss over a period of 8 days. Three mineral oils were also examined for their adduct forming ability and one was selected for adduct formation and loss over a period of 8 days. In addition, fractions derived from the same oil containing 2-3- and 4-6 ring aromatic compounds were applied to mouse skin in a non-carcinogenic oil vehicle and adduct levels were observed over an 8-day period. It was found that PAHs that had no mutagenic, initiating or carcinogenic activity and those that had mutagenic activity in bacteria but no initiating activity in mouse skin failed to produce DNA adducts in mouse skin. Two of the three PAHs with initiating activity and both complete carcinogens produced clear evidence of adduct formation, the adduct levels produced by complete carcinogens being 100 1000 times greater than those produced by initiators. Examination of adduct formation and loss with the carcinogenic PAHs benzo[a]pyrene and 5-methylchrysene over an 8-day period showed a peak at 24 h and an apparent two-phase process of adduct loss. It is suggested that the first steep loss was due to DNA repair and that the more gradual subsequent loss was probably due to epidermal hyperplasia and desquamation. With the initiator 1, 4-dimethylphenanthrene (three rings) a peak of adduct formation was seen at 2 days and adduct levels were not reduced much by 8 days. This suggested that, with initiators, adduct formation and repair may be spread over a longer period than with complete carcinogens. With the whole oils, clear evidence of adduct formation was seen with both a carcinogenic non solvent-refined oil and with a non-carcinogenic residual oil. The level of adduct formation with the residual oil, however, was much lower than with the carcinogenic oil. When adduct formation by the carcinogenic oil was examined over 8 days, the pattern of adduct formation and loss was similar to that of a tumour initiator rather than a complete carcinogen. Peak adduct levels on the diagonal of the thin-layer chromatography (TLC) plates seemed to occur at 1 and 4 days after treatment, with no clear reduction after 8 days. From examination of adducts formed by the 2-3-ring and 4-6-ring aromatic fractions, it appeared that the main adduct spots produced by the carcinogenic oil were due to the 2-3-ring aromatic components of the oil. Adduct spots near the vertical axis of the TLC plates were also seen with the 2-3-ring and 4-6-ring fractions. The relevance of these spots is uncertain, but if they truly represent adducts, the findings suggest that they are due mainly to 4-ring PAHs. The studies suggest that the activity of carcinogenic oils is largely due to substituted 3- and 4-ring polycyclic aromatic compounds and that more attention should be paid to substituted 3-ring compounds in predicting the carcinogenic potential of oils from analytical data. PMID- 10797469 TI - Increased production of interferon-gamma, but not IL-4 mRNA, by streptozotocin in the popliteal lymph node assay. AB - The popliteal lymph node (PLN) assay has been proposed as a tool to predict systemic autoimmune reactions induced by medicinal products and chemicals, the mechanisms of which are poorly understood. To determine whether PLN responses involved Th1 or Th2 cell control, or both, the effects of streptozotocin (STZ), a prototypic immunotoxic compound, were analysed on the production of interferon gamma (IFN-gamma) and interleukin-4 (IL-4) mRNA by lymph node cells after injection into the hind footpad of C57 BL/6 mice. Streptozotocin induced a dramatic increase in IFN-gamma mRNA production, which correlated with PLN responses as evidenced by augmented weight and cellularity indices. No effect on IL-4 mRNA synthesis was noted. These results suggest that a Th1 response is involved in the PLN response to STZ. PMID- 10797470 TI - Neurotoxicity of manganese chloride in neonatal and adult CD rats following subchronic (21-day) high-dose oral exposure. AB - The purpose of this study was to evaluate the relative sensitivity of neonatal and adult CD rats to manganese-induced neurotoxicity. Identical oral manganese chloride (MnCl(2)) doses (0, 25, or 50 mg kg(-1) body wt. day(-1)) were given to neonatal rats throughout lactation (i.e. from postnatal day (PND) 1 through 21) and to adult male rats for 21 consecutive days. The MnCl(2) doses administered to neonates were ca. 100-fold higher than those resulting from the consumption of an equivalent volume of rat's milk. Rats were assessed using similar behavioral and neurochemical evaluations. Several statistically significant changes occurred in Mn-exposed rats relative to control animals. Neonates given the high dose of MnCl(2) had reduced body weight gain. An increased pulse-elicited acoustic startle response amplitude was observed in neonates from both MnCl(2) treatment groups on PND 21. Increased striatal, hippocampal, hindbrain and cortical Mn concentrations were observed in all Mn-exposed neonates on PND 21. Increased hypothalamic and cerebellar Mn concentrations were also observed on PND 21 in neonates from the high-dose group only. Increased striatal, cerebellar and brain residue Mn concentrations were observed in adult rats from the high-dose group. Increased striatal dopamine and 3,4-dihydroxyphenylacetic acid levels were observed only in PND 21 neonates from the high-dose group. No treatment-related changes were observed in clinical signs, motor activity (assessed in neonates on PND 13, 17, 21 +/- 1 and in adults), passive avoidance (assessed in neonates on PND 20 +/- 1 and in adults) or neuropathology (assessed in PND 21 neonates only). The results of our experiment suggest that neonates may be at greater risk for Mn induced neurotoxicity when compared to adults receiving similar high oral levels of Mn. PMID- 10797471 TI - Regulation of nitric oxide synthesis in the liver. AB - Nitric oxide signalling during the past two decades has been one of the most rapidly growing areas in biology. This simple free radical gas can regulate an ever-growing list of biological processes. Here the regulation of NO synthesis in the liver is reviewed. The biogenesis of nitric oxide (NO) is catalysed by nitric oxide synthases (NOS). These enzymes catalyse the oxidation of one of the guanidino nitrogens of l-arginine by molecular oxygen to form NO and citrulline. Three NOS have been identified: two constitutive (cNOS: type 1 or neuronal and type 3 or endothelial) and one inducible (iNOS: type 2). As to the liver, cNOS activity is normally detectable in Kupffer cells, whereas no cNOS is ever encoded in hepatocytes. However, hepatocytes, Kupffer and stellate cells (the three main types of liver cells) are prompted to express an intense iNOS activity once exposed to effective stimuli such as bacterial lipopolysaccharide and cytokines. This review is focused mainly on two aspects: regulation of NOS activity and expression by endogenous and exogenous compounds. Because NO production has beneficial and detrimental effects, understanding the molecular mechanisms that govern NOS is critical to developing strategies to manipulate NO production in liver diseases. PMID- 10797473 TI - Postnatal evolution of supernumerary ribs in rats after a single administration of sodium salicylate. AB - Radiographs were used to follow the postnatal evolution of 14th ribs in rat pups. Initially, 30 pregnant female rats were randomly distributed into two groups receiving 0 or 300 mg kg(-1) sodium salicylate on day 9 of pregnancy. In the treated group, adverse effects were noted on body weight changes and food consumption during the 2 days following dosing. At birth, a high majority of pups had extra ribs at the 300 mg kg(-1) dose. Radiographs done on postnatal days 1, 6, 14, 28 and 54 showed a reduction in the incidence of rudimentary ribs only, whereas extra ribs, often associated with 27 presacral vertebrae, had the same incidence from birth to adult stage. Furthermore, extra ribs seemed to exhibit similar growth evolution to the other thoracic ribs. This work helps to clarify the postnatal evolution of supernumerary ribs because it was performed on the same animals from birth to adult stage, showing that the reversibility was related to rib length and, in consequence, concerned the rudimentary ribs only. The coexistence of additional presacral vertebrae primarily with extra ribs suggests that both kinds of supernumerary ribs (rudimentary and extra) might be different phenomena and could be considered separately in developmental toxicology studies. PMID- 10797472 TI - Study of quinalphos (an environmental oestrogenic insecticide) formulation (Ekalux 25 E.C.)-induced damage of the testicular tissues and antioxidant defence systems in Sprague-Dawley albino rats. AB - The effect of quinalphos (O-O-diethyl-O-[quinoxalinyl-(2)-thionophosphate]), an environmental oestrogenic organophosphorus insecticide pollutant, was studied on Sprague-Dawley albino rats at doses of 250 and 500 microgram kg(-1) body wt. i.p. for 3, 8 and 15 days, respectively. After the treatment with quinalphos there was an increase in the lipid peroxides (as measured by manoldialdehyde production) and a decrease in the total lipid content for the testicular membrane. The effects were more pronounced at the low doses than at the higher doses, indicating that some physiological defence mechanisms were in operation at higher doses. The free-radical-scavenging enzymes (superoxide dismutase, catalase and glutathione peroxidase) showed significantly higher activity at high dose in comparison to their activities at low-dose treatment. Glutathione content was gradually reduced after quinalphos treatment, both in low and high doses. Histomorphohological studies showed that after the low-dose treatment shrinkage of the tubular diameter and testicular atrophy leading to degenerative changes in the germinal epithelium were observed. But at high dose, gradual recovery of various germ cell layers and significant expansion of seminiferous tubules of Sprague-Dawley rat testes were found; the latter is generally believed to be a useful index of testicular activity through the hypothalamo-hypophyseal-gonadal axis. The present study indicates that quinalphos caused damage and degeneration of the testicular tissues due to free-radical-mediated lipid peroxidation at low doses. We have also demonstrated that, in response to the damage, an endogenous antioxidant enzyme defence system became operative at the higher dose of treatment. PMID- 10797474 TI - Evidence for zinc protection against 2,5-hexanedione toxicity by co-exposure of rats to zinc chloride. AB - The protective role of zinc against the toxic effects of 2, 5-hexanedione (2,5 HD), the main neurotoxic metabolite of n-hexane, was investigated by studying the interference of zinc on the toxicokinetics of 2,5-HD. Six groups of Wistar rats were exposed for 3 days to diets containing 2,5-HD, zinc chloride and 2,5-HD+zinc chloride. The amounts of pyrroles and free and total 2,5-HD in urine were determined using Ehrlichs's reagent and gas chromatography/flame ionization detection, respectively. The results show that after the first day of co-exposure (ZnCl(2)+2,5-HD) there was a significant decrease in the excretion of pyrroles and free 2, 5-HD in rats exposed to the chemical mixture when compared to the pyrroles and free 2,5-HD excreted in rats exposed to 2,5-HD alone. However, no significant decrease was observed in the urinary excretion of total 2,5-HD (free 2,5-HD + preformed 2,5-HD). Suggestions are made about the role played by this metal ion in inhibiting pyrrole formation. PMID- 10797475 TI - Effects of intermittent exposures of aflatoxin B(1) on hepatic and testicular glutathione S-transferase in rats. AB - Glutathione S-transferase (GST) plays a major role in the detoxification of the potent hepatocarcinogen aflatoxin B(1) (AFB(1)). This study evaluated the effects of intermittent exposures to AFB(1) on hepatic and testicular GST in rats. Male Fischer 344 rats were fed diets containing AFB(1) (0, 0.01, 0.04, 0.4 and 1.6 ppm) intermittently at 4-week intervals up to 20 weeks. The control animals were fed an AFB(1)-free NIH-31 diet. Rats consuming diets with 0.01 ppm AFB(1) did not show the induction of hepatic or testicular GST activity. Intermittent exposures to AFB(1) at concentrations of 0.04-1.6 ppm significantly increased the GST activities. The increase of the enzyme activity was proportional to the dose and length of AFB(1) exposure. PMID- 10797476 TI - Characterization in mice of the immunological properties of five allergenic acid anhydrides. AB - Occupational exposure to certain acid anhydrides, including trimellitic anhydride (TMA), maleic anhydride (MA), phthalic anhydride (PA), hexahydrophthalic anhydride (HHPA) and methyltetrahydrophthalic anhydride (MTHPA), has been associated with the development of respiratory allergy or asthma. There is considerable debate about the mechanisms through which such chemicals may cause respiratory sensitization, particularly concerning a universal requirement for specific IgE antibody. Despite the controversy regarding an obligatory role for IgE, there is a growing consensus that chemical respiratory hypersensitivity is associated with the selective development of T lymphocytes with a type 2 (Th2) phenotype. In the current investigations we have characterized in mice the nature of immune responses provoked by prolonged topical exposure to five acid anhydrides. Under application conditions where similar overall immunogenicity was achieved, we have compared cytokine responses induced by PA, MA, HHPA and MTHPA with those provoked by concurrent exposure to TMA or to the reference contact allergen 2, 4-dinitrochlorobenzene (DNCB). Lymph node cells (LNC) draining the site of topical exposure to DNCB invariably expressed high levels of the type 1 cytokines interferon-gamma (IFN-gamma) and interleukin-12 (IL-12), but only low levels of the type 2 cytokines interleukin-4 (IL-4) and interleukin-10 (IL-10). In each experiment, TMA-activated LNC displayed the converse, type 2, phenotype of cytokine production. The other acid anhydrides in each case provoked a type 2 cytokine secretion profile, with comparable IL-10 expression but somewhat less vigorous IL-4 production compared with that observed following exposure to the reference respiratory allergen TMA. In every experiment relatively low levels of IFN-gamma and IL-12 were elaborated by acid anhydride-activated LNC, with the exception of PA-stimulated LNC that displayed increased amounts of IL-12 in comparison with other acid anhydrides. Thus, prolonged topical exposure of mice to five different acid anhydrides in each case resulted in the development of a predominantly Th2-type cytokine secretion phenotype, consistent with the ability of these materials to provoke asthma and respiratory allergy through a type 2 (possibly IgE-mediated) mechanism. Taken together with the results of previous investigations with a wider range of chemical allergens, these data suggest that induced cytokine secretion patterns or 'fingerprints' allow discrimination between contact and respiratory allergens and consequently represent a suitable approach to prospective evaluation of respiratory sensitization hazard. PMID- 10797477 TI - In vivo and in vitro studies on the genotoxicity of cadmium chloride in mice. AB - The genotoxic effect of cadmium chloride was evaluated in chromosomes of experimental mice using in vivo and in vitro studies. In vivo the induction of micronuclei, sister chromatid exchange in mouse bone marrow and chromosomal aberrations in both somatic and germ cells was investigated. Doses 1.9, 5.7 and 7.6 mg kg(-1) body wt. (single i.p. treatment) induced a significant and dose dependent increase in the percentage of polychromatic erythrocytes with micronuclei. Such a percentage reached 2.1% with the highest tested dose, compared with 0.57% for the control (non-treated) and 2.2% for mitomycin c as the positive control. The dose of 1.9 mg kg(-1) body wt. had no significant effect with respect to sister chromatid exchange (SCE) but the doses of 5.7 and 7.6 mg kg(-1)body wt. increased the frequency of SCEs significantly. The frequency of SCE reached 7.35 +/- 0.26 per cell after treatment with the highest tested dose, which is a less than twofold increase compared with the control frequency of 4.6 +/- 0.42 per cell. However mitomycin c induced a much higher effect (12.1 +/- 0.73). Cadmium chloride also induced a significant increase in the percentage of chromosomal aberrations in mouse bone marrow at the doses of 5.7 and 9.5 mg kg( 1) body wt. (single i.p. treatment). The effect is a function of cadmium chloride concentration. Moreover, cadmium chloride induced its maximum effect concerning the induction of chromosomal aberrations in mouse bone marrow cells 24 h after treatment, compared with 12 and 48 h. In germ cells, chromosomal aberrations were observed in mouse spermatocytes 12 days post-treatment with the dose of 5.7 mg kg(-1) body wt. Moreover, a pronounced reduction in the number of spermatocytes was observed after administration of cadmium chloride (0.9, 1.9 and 5.7 mg kg(-1) body wt.) In in vitro studies, the three tested concentrations of 10, 15 and 20 microgram ml(-1) cadmium chloride induced a statistically significant increase in the frequency of SCEs in cultured mouse spleen cells. The concentrations of 15 and 20 microgram ml(-1) also induced chromosomal aberrations in mouse spleen culture. The ability of vitamin C (l-ascorbic acid) to minimize the incidence of chromosomal aberrations induced by cadmium chloride in cultured mouse spleen cells was investigated. Vitamin C at the concentrations of 3 and 6 microgram ml( 1) significantly minimized the percentage of aberrant cells induced by cadmium chloride. PMID- 10797478 TI - Effect of methanol intoxication on free-radical induced protein oxidation. AB - Oxygen free radicals are generated during methanol-induced liver injury, as was shown for ethanol. The effect of methanol intoxication (6 g kg(-1) body wt.) on protein modification in the liver of rats was investigated. Electron spin resonance determination indicated an increase in the free radical signal 6 and 12 h after intoxication. After 7 days of treatment, the contents of malondialdehyde and carbonyl groups in proteins were significantly increased. The level of amino groups and sulphydryl groups and the amount of tryptophan in proteins were decreased, whereas the amount of bi-tyrosine was increased significantly. Changes in protein structure resulted both from free radical action and formaldehyde generation during methanol intoxication. PMID- 10797479 TI - Glial dependent survival of neurons in Drosophila. AB - According to the classical model of insect neurogenesis, neuron fate and survival is determined largely by cell autonomous mechanisms with no requirement for cell cell interactions to control the total number of neurons. In a recent paper by Booth et al.,(1) however, the central tenet of this model has been called into question. Using a combination of mutations and targeted glial ablation, this paper shows that, contrary to common thinking, neuron survival in the embryonic nervous system of Drosophila is dependent upon normal glial function. This surprising result suggests that insect neurogenesis may have more in common with vertebrate neurogenesis than previously thought. PMID- 10797480 TI - The real Dorian Gray mouse. AB - Genetic variants with greatly extended lifespan are proving invaluable in uncovering signal transduction pathways that influence the rates of normal ageing. These studies have so far been confined to invertebrate models such as Caenorhabditis elegans and Drosophila, but there has been much speculation as to whether a similar approach could be applied to mammals. The recent publication of results on a mouse strain, mutant in a gene encoding the signaling molecule p66(shc), gives cause for optimism. The mutation renders the mouse resistant to the action of oxygen radical generators and appears to increase mean lifespan by 30%. This approach may provide a boost for the modeling of human age-related diseases. PMID- 10797481 TI - Dysregulation of lymphocyte proliferation by chromosomal translocations and sequential genetic changes. AB - Enzymatically mediated rearrangement of Ig and T-cell receptor genes is essential for generating the huge molecular repertoire of the mammalian immune system, but it also carries a danger for the organism in the form of high risk zones for illegitimate juxtaposition of DNA from other areas of the genome. Translocation dependent activation of oncogenes, transcription factors or developmental genes can trigger the development of neoplasia in a lineage-specific fashion. These events are not sufficient for tumorigenesis, however, since some of the most prominent tumor-associated translocations, such as Ig/myc and Ig/bcl-2, have been detected in normal individuals who did not develop tumors. Tumor development must, therefore, require subsequent genetic changes. Among them, the increased expression of genes that protect against apoptosis or, alternatively, mutations that cripple apoptosis-activating genes play a prominent role. Some of the translocations associated with T-cell leukemia, myeloid leukemia, and a variety of sarcomas act by generating fusion proteins. The participating genes encode transcription factors and/or developmental regulators. Fusion protein-expressing cells may serve as targets for specific interference with abnormal signaling pathways or for targeted immune attack. Using PCR to detect cells carrying such translocations is useful for tumor diagnosis, prognosis, and choice of therapy. PMID- 10797482 TI - Germ cell suicide: new insights into apoptosis during spermatogenesis. AB - Mature sperm are the product of a precisely regulated developmental sequence in which germ cell proliferation, differentiation, self-renewal and apoptosis are carefully controlled. The control of germ cell apoptosis during spermatogenesis is especially important. It is mediated by signals derived from the Sertoli cells with which each germ cell is closely associated, as well as by signals originating outside the testis. A greater understanding of these signals is emerging from studies of the spermatogenic defects of genetically modified animals. In particular, the intracellular signaling cascades which ultimately determine germ cell fate are being illuminated by recent studies of the Bcl-2 protein family. This review summarises the crucial role which stringently regulated apoptosis plays in the production of male gametes. PMID- 10797483 TI - Homeobox genes and gut development. AB - The gut of vertebrates exhibits a common anteroposterior regional differentiation. The role of homeobox genes in establishing this pattern is inferred by their sites of expression. It is suggested that the primary source of positional information is in the endoderm, which subsequently establishes a 'dialogue' with the surrounding visceral layer of the lateral plate mesoderm. This results in the anatomical and physiological specialization of the adult gut. PMID- 10797484 TI - Ubiquitin-mediated proteolysis: biological regulation via destruction. AB - The ubiquitin proteolytic system plays an important role in a broad array of basic cellular processes. Among these are regulation of cell cycle, modulation of the immune and inflammatory responses, control of signal transduction pathways, development and differentiation. These complex processes are controlled via specific degradation of a single or a subset of proteins. Degradation of a protein by the ubiquitin system involves two successive steps, conjugation of multiple moieties of ubiquitin and degradation of the tagged protein by the 26S proteasome. An important question concerns the identity of the mechanisms that underlie the high degree of specificity of the system. Substrate recognition is governed by a large family ubiquitin ligases that recognize the substrates, bind them and catalyze/facilitate their interaction with ubiquitin. PMID- 10797485 TI - Mechanisms leading to uniparental disomy and their clinical consequences. AB - Uniparental disomy (UPD) refers to the situation in which both copies of a chromosome pair have originated from one parent. In humans, it can result in clinical conditions by producing either homozygosity for recessive mutations or aberrant patterns of imprinting. Furthermore, UPD is frequently found in conjunction with mosaicism for a chromosomally abnormal cell line, which can also contribute to phenotypic abnormalities. Investigations into the mechanisms by which UPD may arise have helped to expand our general awareness of the impact of chromosomal abnormalities and chromosomal mosaicism in normal human development. Specifically, it appears that errors in the transmission of a chromosome from parent to gamete and during early somatic cell divisions are remarkably common but that embryo and cell selection during early embryogenesis help to ensure the presence of a numerically balanced chromosome complement in the developing fetus. UPD is also likely to occur within a portion of cells in all individuals simply as a consequence of somatic recombination occurring during mitotic cell divisions. This can be an important step in cancer development as well as a contributing factor to other late onset diseases. This review summarizes mechanisms by which UPD may arise and their associated clinical consequences. PMID- 10797486 TI - hedgehog and wing development in Drosophila: a morphogen at work? AB - In Drosophila, Hedgehog (Hh) is a key regulator of limb development and activates decapentaplegic (dpp), a gene encoding a TGFbeta-related factor that controls growth and patterning of the limbs. During wing development, Hh also has morphogen-like Dpp-independent functions, controlling the morphogenesis of the central part of the wing through the activation of the evolutionarily conserved transcription factors encoded by the iroquois and collier genes. The ways in which Hh forms an activity gradient to lay the basis of patterning of the adult wing are described here. As the signal transduction pathway of Hh is strongly conserved during evolution and human Hh may be implicated in congenital diseases and cancers, these observations provide important advances which may help in understanding the function of Hh proteins in normal and pathological development and tumourigenesis in humans. PMID- 10797487 TI - Cytotoxicity and cytotoxic molecules in invertebrates. AB - Although lacking the components that characterize the acquired immunity systems of vertebrates, invertebrates nevertheless possess effective general innate immune mechanisms which exhibit striking parallels with those of vertebrates. These innate immune systems include both cellular and humoral elements. Invertebrate phagocytes synthesize both oxygen-dependent and oxygen-independent molecules to combat infectious agents. Cytotoxic substances employed by invertebrates include reactive intermediates of oxygen and nitrogen, antimicrobial peptides, lectins, cytokine- and complement-like molecules, and quinoid intermediates of melanin. The signal transduction pathways that are involved in mediating the production of these substances appear to be very similar among animal species, suggesting a common ancestral origin for the innate immune systems. PMID- 10797488 TI - Prokaryotic and eukaryotic chromosomes: what's the difference? AB - It is widely held that the profound differences in cellular architecture between prokaryotes and eukaryotes, in particular the housing of eukaryotic chromosomes within a nuclear membrane, also extends to the properties of their chromosomes. When chromosomal multiplicity, ploidy, linearity, transcriptional silencing, partitioning, and packaging are considered, no consistent association is found between any of these properties and the presence or absence of a nuclear membrane. Some of the perceived differences can be attributed to cytological limitations imposed by the small size of bacterial nucleoids and the arbitrary choice of representative organisms for comparison. We suggest that the criterion of nucleosome-based packaging of chromosomal DNA may be more useful than the prokaryote/eukaryote dichotomy for inferring the broadest phylogenetic relationships among organisms. PMID- 10797489 TI - Medakafish as a model system for vertebrate developmental genetics. AB - Several teleosts, such as the zebrafish and the medakafish or medaka (Oryzias latipes), are used as vertebrate model systems in various fields of biology. The medaka is suitable for use in genomic studies because of its small genome size. Moreover, our recent results of small-scale mutagenesis in the medaka indicate that it is possible to identify mutations, the phenotypes of which could not be found in zebrafish mutants obtained by large-scale mutagenesis. An example is Oot (One-sided optic tectum), a maternal-effect mutation. In the Oot phenotype, bilateral symmetry is broken in the optic tectum in the early developmental stages, and either the left or right morphology is duplicated on both sides. Medaka inbred strains can be produced and used to study quantitative traits in vertebrate development. Data presented support the use of medaka as another important fish model for the study of vertebrate developmental genetics. PMID- 10797490 TI - Effects of androgens on telomerase activity in normal and malignant prostate cells in vitro. AB - BACKGROUND: Recent studies have shown that sex hormones regulate telomerase activity in endometrium and breast tissues. The present study was designed to clarify the effects of androgen on telomerase activity in normal and malignant prostate cells. METHODS: Androgen-sensitive (LNCaP) and -independent (TSU-Pr1 and DU145) prostate cancer cell lines and normal prostate cells including basal cells were cultured in the presence or absence of 5alpha-dihydrotestosterone (DHT). RESULTS: Prostate cancer cell lines exhibited high telomerase activity, and normal prostate cells showed low activity. Short or prolonged androgen deprivation reduced telomerase activity in LNCaP cells, and DHT induced telomerase activity at the G(1) phase of the cell cycle. DHT did not modulate telomerase activity in TSU-Pr1, DU145, and normal cells. CONCLUSIONS: LNCaP cells had an androgen-dependent pathway to activate telomerase, whereas TSU-Pr1 and DU145 cells as well as normal prostate cells had an androgen-independent pathway. These findings suggest that the regulatory mechanism of telomerase varies during the progression of prostate cancers. PMID- 10797491 TI - Prostasomes: current concepts. AB - BACKGROUND: Prostasomes are membranous vesicles secreted by prostate gland, and they contain large amounts of cholesterol, sphingomyelin, calcium, and several enzymes. Prostasomes are involved in a number of biological functions. At ejaculation, these prostasomes are expelled with prostate secretions and are to be found in the seminal plasma as seminal prostasomes, which facilitate sperm function in various ways. METHODS: In this review, we discuss the structural and functional role of prostasomes, the various enzyme systems associated with these vesicles, and the biological role prostasomes play in male reproduction. RESULTS AND CONCLUSIONS Prostasomes are pluripotent and well-organized organelles secreted by the prostate gland. Prostasomes are ascribed to have many physiologiocal functions, the primary function being enhancement of sperm capacity. The several enzyme systems, small signaling molecules, and neuroendocrine markers associated with prostasomes reveal the complex nature of these vesicles in regulating sperm viability and vitality. The functional significance of these molecules that regulate complex pathways in these small vesicles is still a matter of dogma. Critical evaluation of the biological processes associated with prostasomes might be helpful in modeling new contraceptive agents, improving the techniques of in vitro fertilization, and in furthering our understanding and treatment of male factor infertility. PMID- 10797492 TI - Galactosyl receptor, a cell surface C-type lectin of normal and tumoral prostate epithelial cells with binding affinity to endothelial cells. AB - BACKGROUND: The mechanism of bone metastasis of prostate cancer involves the interaction of cell surface receptor(s) on cancer cells with ligand(s) on bone marrow endothelial cell surfaces. The rat galactosyl receptor gene generates two mRNA species by differential splicing: one species encodes a protein identical to the minor form of hepatocyte asialoglycoprotein receptor and displays a galactose/N-acetyl-galactosamine-recognition domain; the other encodes a protein with identical intracellular and transmembrane domains but with a different extracellular domain lacking the carbohydrate-recognition domain (CRD). Both proteins appear to coexist as a heterooligomer on the surface of normal mouse, rat, and human prostate epithelial cells and human prostate cancer cells, including the PC-3 cell line. The CRD of galactosyl receptor mediates adhesion of normal and tumoral prostate cells to the surfaces of a human bone marrow endothelial cell line. The use of inhibitors targeting the CRD would be very valuable in hindering the binding of prostate cancer cells to endothelial cells, thus decreasing the incidence of hematogenous metastasis to bone. METHODS: Molecular biology, immunohistochemistry, flow cytometry, and a cell aggregation assay were used to determine the expression and role of the galactosyl receptor in cell adhesion. RESULTS: Immunoblotting experiments demonstrated that each component of the heterooligomer has a mass of 54 kDa, ascribed in part to associated carbohydrates. An oligonucleotide probe showed the presence of both galactosyl receptor forms in rat prostate and testis, but not in liver, kidney, and spleen. Antibodies to the CRD and a segment of the nonhomologous extracellular domain of the galactosyl receptor blocked cell adhesion to endothelial cell monolayers. CONCLUSIONS: The galactosyl receptor provides a valuable target for the development and use of synthetic ligands capable of disrupting endothelial cell-prostate cancer cell interaction, the first step in prostate cancer bone metastasis. PMID- 10797493 TI - Vascular endothelial growth factor-A expression in the rat ventral prostate gland and the early effects of castration. AB - BACKGROUND: Blood flow to the rat ventral prostate gland is drastically reduced during the very early period after castration, and this reduction coincides with the appearance of striking degenerative changes within the prostatic vascular system. These early effects on the prostate vascular system are likely to be important for the subsequent regression of the ventral prostate that occurs in response to castration. Since the endothelial cells of the ventral prostate do not express androgen receptor protein (AR), we proposed that these early effects might be indirectly mediated by changes in the local expression of vascular regulatory factors. In order to evaluate whether vascular endothelial growth factor-A (VEGF-A) might be among the primary mediators of these effects, we measured expression of VEGF-A mRNA and protein in the rat ventral prostate gland prior to and within the first 3 days after castration. METHODS: Ventral prostate tissues were obtained from control (unoperated) rats, sham-operated rats, or rats at sequential daily intervals (1-3 days) after castration. A quantitative RNase protection assay and a comparative RT-PCR assay were used to evaluate the extent to which the expression of VEGF-A mRNA in the ventral prostate was affected by castration. In situ immunohistochemistry, using an anti-VEGF-A antibody, was performed to localize VEGF-A protein in the various cells of the tissue. Western blot analysis and a quantitative ELISA assay using anti-VEGF-A antibodies were performed to determine how VEGF-A protein expression in the rat ventral prostate was affected by castration. RESULTS: Results of VEGF-A mRNA analysis in the rat ventral prostate gland during the first 3 days after castration showed a biphasic change characterized by a transient reduction of VEGF-A mRNA expression (by approximately 50%) on the second day after castration that was restored to higher than control levels by the third day after castration. Immunohistochemical analysis for VEGF-A in control and castrated ventral prostates showed that the prostatic epithelial and smooth muscle cells were the major source of VEGF-A expression in this tissue. Quantitative analysis of VEGF-A protein expression by Western blot and ELISA methods confirmed a biphasic change in the expression of the polypeptide that correlated well with the results of the mRNA analyses. CONCLUSIONS: VEGF-A expression in the ventral prostate gland of the Sprague Dawley rat is downregulated on the second day after castration but returns to control levels by the third day after castration. Since critical changes in the ventral prostate vascular system are already evident by 1 day after castration, we believe that these findings indicate that VEGF-A is not likely to be the critical or sole mediator of the early effects of castration on the vascular system of the rat ventral prostate gland. PMID- 10797494 TI - Prostatic luminal cell differentiation and prostatic steroid-binding protein (PBP) gene expression are differentially affected by neonatal castration. AB - BACKGROUND: Although normal prostatic development is androgen-dependent, the prostate continues to grow in the neonate despite castration. However, the manner in which neonatal growth of the prostate occurs, in the absence of the testis, remains largely unknown. The purpose of this study was to examine the differentiation of prostatic epithelial cells after neonatal castration. METHODS: Immunohistochemistry was utilized to detect the expression of differentiation products: basal-cell cytokeratin (CK 5), luminal-cell cytokeratin (CK 18), and prostatic steroid-binding protein (PBP), a ventral prostate-specific marker indicative of secretory function in luminal cells. The reverse transcription polymerase chain reaction was used to detect transcription products of the three polypeptide subunits of PBP, designated C1, C2, and C3. Rats were castrated on day 5 after birth, and ventral prostates were collected on day 14. Dihydrotestosterone was injected (100 microg/animal every 2 days) in castrated animals to determine if PBP expression could be initiated by androgen. RESULTS: Although no major effects of castration were detected on the differentiation of stromal or basal cells (which differentiate prior to day 5), castration had a pronounced effect on luminal-cell differentiation. Castration inhibited PBP protein expression, but did not affect the expression of luminal-cell cytokeratin (CK 18) protein. Furthermore, castration reduced C1, C2, and C3 transcription. Androgen replacement to castrated animals allowed for the initiation of PBP expression, although its onset was delayed. CONCLUSIONS: These observations indicate that the testis is not necessary for prostatic luminal-cell differentiation, but is necessary for full expression of luminal-cell secretory phenotype. Furthermore, our study suggests that factors of testicular origin, in addition to androgen, are needed for proper timing of PBP expression. This investigation establishes that the cytological and the physiological differentiation of the rat prostate are differentially regulated. PMID- 10797495 TI - Evidence of functional ryanodine receptor involved in apoptosis of prostate cancer (LNCaP) cells. AB - BACKGROUND: Very little is known about the functional expression and the physiological role of ryanodine receptors in nonexcitable cells, and in prostate cancer cells in particular. Nonetheless, different studies have demonstrated that calcium is a major factor involved in apoptosis. Therefore, the calcium regulatory mechanisms, such as ryanodine-mediated calcium release, may play a substantial role in the regulation of apoptosis. METHODS: We assessed the presence of such functional receptors in LNCaP prostate cancer cells, using fluorimetric measurements of intracellular calcium and expression assays of mRNA encoding ryanodine receptors. RESULTS: We show here that LNCaP cells responded to caffeine, a ryanodine receptor agonist, by mobilizing calcium. Another ryanodine receptor agonist, 4-chloro-m-cresol, had a similar effect and promoted calcium release. These effects were inhibited by pretreatment with ryanodine or thapsigargin. In addition to a calcium release, caffeine was able to produce a calcium entry blocked by nickel. We used a reverse transcription-polymerase chain reaction assay to investigate the expression of ryanodine receptors in LNCaP cells. Two types of ryanodine receptor mRNAs were expressed in LNCaP cells: RyR1 and RyR2 mRNAs. Finally, we show that ryanodine receptor activation by caffeine slightly stimulates apoptosis of prostate cancer cells, and that the inhibition of these receptors by ryanodine protects the cells against apoptosis. CONCLUSIONS: The combination of results showed that LNCaP cells, derived from a human prostate cancer, express functional RyRs able to mobilize Ca(2+) from intracellular stores and which might control apoptosis. PMID- 10797496 TI - Screening and early hormonal treatment of prostate cancer are accumulating strong evidence and support. AB - BACKGROUND: I review the data published during the last 5 years on the effects of early treatment of prostate cancer on survival. METHODS: Data from prospective and randomized studies as well as from population-based studies are presented. RESULTS: Two studies (European Organization for Research and Treatment of Cancer and Radiation Therapy Oncology Group) in stage T3 disease have shown that long term (3 years or indefinite, respectively) androgen blockade prolongs life in patients receiving androgen blockade in addition to radiotherapy compared to radiotherapy alone. In the UK Medical Research Council study, androgen blockade at diagnosis of locally advanced or asymptomatic patients decreased cancer specific death by 21% compared to delayed treatment. A 69% decrease in prostate cancer death was observed in the Quebec Randomized Prostate Cancer Screening Study. Population-based studies in Sweden and Denmark have shown that 62% and 63%, respectively, of patients diagnosed with localized prostate cancer will die from the disease if not treated immediately. Decreases in prostate cancer death rate of 6.3-23% have been found between 1991-1997 in the US and Canada, respectively. CONCLUSIONS: Treatment of localized disease has been shown in all the available randomized studies to cause a marked decrease in prostate cancer death. Simple use of the available screening procedures and treatments for localized prostate cancer could cause a dramatic decrease in prostate cancer death. PMID- 10797497 TI - Screening and early treatment of prostate cancer are accumulating strong evidence and support. PMID- 10797499 TI - Analysis of cyclin-dependent kinase inhibitor expression and methylation patterns in human prostate cancers. AB - BACKGROUND: Downregulation of genes which negatively control cell cycle progression represents a possible mechanism for prostate tumorigenesis. We examined the expression levels of the p16, p15, p14, and retinoblastoma susceptibility (RB) genes in primary prostate cancers and human prostate cancer cell lines, and correlated this with the DNA methylation levels of two loci in p16. METHODS: The mRNA levels of p16, p15, and p14 were examined by reverse transcriptase-PCR (RT-PCR). DNA methylation of the p16 5' CpG island was determined by bisulfite genomic sequencing, while methylation of exon 2 shared by the p16 and p14 genes was measured by a quantitative bisulfite-based technique, methylation-sensitive single-nucleotide primer extension (Ms-SNuPE). RB protein levels were assessed by immunohistochemical staining of histologic sections of normal and tumor prostate tissues, using a monoclonal antibody (mAB). RESULTS: Overexpression of p16 mRNA was found in 6/9 (67) of prostate tumors compared to the adjacent normal prostate, whereas elevated p14 and p15 levels were only observed in 2/9 (22) and 1/6 (17) of prostate cases, respectively. There was no statistically significant association of grade (P = 0.18) and stage (P = 1.00) of prostate cancer to the elevated p16 levels in the tumors. The p16 5' CpG island was completely unmethylated in these tissues. In contrast, exon 2 of p16/p14 was methylated in both the tumor and normal adjacent prostates, and was increased in 8/11 (73) of tumors relative to normal tissues. There was no association between p16 overexpression and increased p16/p14 exon 2 methylation in these tumors (P = 1.00). Diminished RB levels in prostate tumors that had upregulated p16 mRNA were found, although absent RB was also detected in tumors without elevated p16 levels. The expression levels of the two genes, RB and p16, were not correlated statistically (P = 0.16). CONCLUSIONS: Our studies show that although the levels of the cell cycle regulators p16, p15, p14, and Rb are altered in prostate cancers, there is no apparent correlation to grade, stage, or any pattern of regulation between the related genes. Exon 2 of p16/p14 is methylated in a majority of prostate tumors compared to the unmethylated upstream 5' region, and may be a potential tumor marker for human prostate cancer. PMID- 10797498 TI - Fluorescence in situ hybridization evaluation of c-myc and androgen receptor gene amplification and chromosomal anomalies in prostate cancer in Japanese patients. AB - BACKGROUND: Oncogene amplification and chromosomal anomalies are found in many solid tumors and are often associated with aggressiveness of cancer. We evaluated the frequency and the association of c-myc and androgen receptor (AR) gene amplification and gain of chromosome 8 or X in prostate cancer in Japanese patients. METHODS: We examined a total of 42 prostate cancer specimens, using fluorescence in situ hybridization (FISH). Dual-labeling hybridization with a directly labeled centromere probe for chromosome 8 or X together with a probe for the c-myc or AR locus was performed. RESULTS: Gain of chromosome 8 was identified in 54.8% of specimens and was associated with Gleason sum and nuclear anaplasia in untreated prostate cancers. c-myc gene amplification was found in 14.3% of specimens. Gain of chromosome X was identified in 42.9% of specimens. AR gene amplification was detected in 0 of 37 untreated prostate cancers, but in 1 of 5 hormone-refractory prostate cancers. CONCLUSIONS: Our results suggest that c-myc and AR gene amplification and gain of chromosome 8 or X may be associated with the development and progression of prostate cancers. These results obtained in Japanese cases are consistent with the results observed in prostate cancer in Western countries. PMID- 10797500 TI - Lung cancer risk of workers in shoe manufacture and repair. AB - BACKGROUND: The occupational lung cancer risk in manufacturing and repair of shoes was studied by pooling of two major case-control studies from Germany. METHODS: Some 4184 incident hospital-based cases of primary lung cancer and 4253 population controls, matched for sex, age, and region of residence were intensively interviewed with respect to their occupational and smoking history. Based on the occupational coding and a free text search, all individuals who had ever worked in shoe manufacturing or repair for at least half a year were identified. Shoemaker-years were calculated as the cumulated duration of working in shoe manufacturing or repair. Odds ratios (OR) and 95% confidence intervals (CI) were calculated via conditional logistic regression. Additional adjustment for smoking and occupational asbestos exposure was used. RESULTS: Seventy-six cases and 42 controls who had ever worked in shoe manufacture or repair (OR = 1.89, 95% CI: 1.29-2.78). After adjustment for smoking, this risk was lowered to 1.69 (95% CI: 1.09-2.62). Further adjustment for asbestos exposure only slightly changed the risk estimates upwards. The smoking adjusted OR in males was 1.50 (95% CI: 0.93-2.41) and 2.91 (95% CI: 0.90-9.44) in females. Logistic regression modeling showed a positive dose-effect relationship between duration of exposure in shoe manufacture and repair and lung cancer risk. The odds ratio for 30 years of exposure varied between 1.98 and 2.24 depending on the model specified. CONCLUSIONS: The study demonstrates an increased lung cancer risk for shoemakers and workers in shoe manufacturing. The risk seems to double after being 30 years in these occupations. PMID- 10797501 TI - Laryngeal and hypopharyngeal cancer and occupational exposure to asbestos and man made vitreous fibers: results of a case-control study. AB - BACKGROUND: The data from a case-control study performed in France between 1989 and 1991 were used to test whether exposure to either asbestos or to man-made vitreous fibers (MMVF) is a risk factor for cancer of the larynx or the hypopharynx. METHODS: This study involved 315 incident cases of laryngeal cancer, 206 cases of hypopharyngeal cancer, and 305 hospital-based controls with other types of cancer, all recruited in 15 hospitals in six French cities. The subjects' past occupational exposure to asbestos and to four types of MMVF (mineral wool, refractory ceramic fibers, glass filaments, and microfibers) was evaluated based on their job history, with the aid of a job-exposure matrix. Odds ratios were calculated with unconditional logistic regression, with adjustment for smoking and drinking levels. RESULTS: Exposure to asbestos resulted in a significant increase in the risk of hypopharyngeal cancer (OR = 1.80, 95% CI: 1.08-2.99) and a nonsignificant increase in the risk of laryngeal cancer (OR = 1.24, 95% CI: 0.83-1.90). Risk was highest for the epilarynx (highest cumulative level of exposure: OR = 2.22, 95% CI: 1.05-4.71). Exposure to mineral wools was of borderline significance for the risk of hypopharyngeal cancer (OR = 1.55, 95% CI: 0.99-2.41), and nonsignificantly associated with the risk of laryngeal cancer (OR-1.33, 95% CI: 0.91-1.95). The risk was again highest for the epilarynx (OR = 1.85, 95% CI: 1.08-3.17). No significant results were observed for the other MMVF. CONCLUSIONS: These results suggest that asbestos exposure increases the risk of epilaryngeal and hypopharyngeal cancers. It is difficult to reach a conclusion about the effects of mineral wools, because nearly all the exposed subjects were also exposed to asbestos. The possible effects of other MMVF were difficult to assess in this study, because of the paucity of exposed subjects. PMID- 10797502 TI - Mortality among rubber chemical manufacturing workers. AB - BACKGROUND: A retrospective cohort mortality study evaluated ischemic heart disease (IHD) among workers in the "rubber chemicals" manufacturing department of a Western New York plant. A previous study at the plant found elevated chest pain and angina among workers in this department. METHODS: Mortality experience of workers employed from 1946-1988 was followed through December 31, 1994. Mortality was compared to U.S. population rates and to local Niagara county rates by using the NIOSH life table analysis system. Poisson regression was used to examine patterns of IHD within the cohort. RESULTS: The standardized mortality ratio (SMR) for IHD among workers in the rubber chemicals department was 1.51 (U.S. rates) and 1.19 (Niagara county rates). Increased mortality from IHD in the rubber chemicals department was most pronounced at younger ages (< 50, SMR = 2.4); workers in a second chemical production department also had an elevated (but not statistically significant) SMR of 1.5 for IHD. CONCLUSIONS: IHD mortality among workers in the rubber chemicals department was elevated, particularly among those under 50 years of age. Potential occupational risk factors for IHD include the rotating shift pattern for employees assigned to two chemical production departments and chemical exposures present in the rubber chemicals department. PMID- 10797503 TI - Hydrocarbons may worsen renal function in glomerulonephritis: a meta-analysis of the case-control studies. AB - BACKGROUND: Previous case-control studies of hydrocarbon exposure in glomerulonephritis may have given inconsistent results because the stage of disease has not been considered. METHODS: The association between stage of disease and degree of previous exposure was studied by a meta-analysis of all published case-control studies and by reviewing all follow-up studies. RESULTS: Odds ratios (OR) for exposure could be calculated for 16 patient groups from 14 case-control studies. After exclusion of four patient groups with 5-17% drop-outs due to death, the mean weighted OR for patient groups with acute or early glomerulonephritis, chronic renal failure, and end-stage renal failure were 0.95, 3.1, and 5.9, respectively. At follow-up, reported in four studies, renal failure was mainly seen in patients with extensive exposure, and improvement was seen only in patients who had discontinued the exposure. CONCLUSIONS: Hydrocarbon exposure in glomerulonephritis is associated with the advancement of the disease and inversely associated with renal function. Early elimination of the exposure may, therefore, prevent the progress of renal failure in many patients. PMID- 10797504 TI - Leukemia in electric utility workers: the evaluation of alternative indices of exposure to 60 Hz electric and magnetic fields. AB - BACKGROUND: Epidemiological studies have inconsistently demonstrated a positive relationship between magnetic and/or electric fields and leukemia. Although exposure to both 60 Hz electric and magnetic fields can be characterized in many ways, to date, risk assessment has been performed by using only a limited number of exposure indices. METHODS: The associations between adult leukemia and indices of electric and magnetic fields were explored within a nested case-control study of 31,453 Ontario electric utility workers. RESULTS: The percentage of time spent above electric field thresholds of 20 and 39 V/m was predictive of leukemia risk after adjusting for duration of employment and the arithmetic mean exposure to both electric and magnetic fields (P<0.05). Duration of employment was strongly associated with an increased risk of leukemia. Those who had worked for at least 20 years, and were in the highest tertiles of percentage of time spent above 10 and 20 V/m had odds ratios of 10.17 (95% CI = 1.58-65.30) and 8.23 (95% CI = 1. 24-54.43), respectively, when compared to those in the lowest tertile. Nonsignificant elevations in risk were observed between indices of magnetic fields and leukemia. CONCLUSIONS: Our results support the hypothesis that electric fields act as a promoting agent in the etiology of adult leukemia. Exposure assessment based on alternate indices of electric and magnetic fields should be incorporated into future occupational studies of cancer. PMID- 10797505 TI - Retinal degeneration in licensed pesticide applicators. AB - BACKGROUND: Retinal degeneration is the leading cause of visual impairment in older adults, but little is known about its relationship to neurotoxic exposures. METHODS: The Agricultural Health Study is a cohort study of licensed pesticide applicators from Iowa and North Carolina. We used cross-sectional data from self administered questionnaires given at enrollment in 1994-1996 to compare pesticide use in 154 applicators who reported retinal degeneration and 17,804 controls. RESULTS: Retinal degeneration was associated with fungicide use (odds ratio = 1.8, 95% confidence interval = 1.3-2.6). This relationship was seen in subgroups defined by state, demographic characteristics, or medical history, as well as in the entire group. Risk increased with cumulative days of fungicide use (P for trend = 0.011) and was greater when application methods involving greater personal exposure were used. Retinal degeneration was also related to use of organochlorine or carbamate insecticides, but these associations were less consistent. Since nearly all applicators used organophosphate insecticides and herbicides, these exposures could not be effectively evaluated. CONCLUSIONS: These results suggest that exposure to some fungicides and insecticides may increase risk of retinal degeneration. PMID- 10797506 TI - Killed on the clock: a population-based study of workplace homicide, 1977-1991. AB - BACKGROUND: This paper describes the epidemiology of workplace homicides in North Carolina, with emphasis on the circumstances. METHODS: Workplace homicide victims were identified by and data were abstracted from the North Carolina medical examiner system. RESULTS: Workplace homicide rates are highest for men, older and self-employed workers, minorities and specific occupations, especially taxi drivers. Robberies, mostly in retail settings, accounted for half of the cases, while 20% were known to involve disputes, the contexts of which differed by sex. Women were most likely to be killed by estranged partners. CONCLUSIONS: Preventive strategies need to address the specific contexts in which workplace homicide occurs, such as retail and taxi robberies, and law enforcement officers interacting with suspects. A workplace response to domestic violence is also needed. Other areas for future research and intervention include environmental modifications, employee screening and training, and identifying more inclusive occupational data sources. PMID- 10797507 TI - Non-fatal occupational fall and slip injuries among commercial fishermen analyzed by use of the NOMESCO injury registration system. AB - BACKGROUND: Fall injuries constitute a significant problem in commercial fishing and such injuries have hitherto not been the subject of closer analysis. METHODS: The distribution and the characteristics of 582 occupational injuries among commercial fishermen are described by using data from an emergency department for the period 1990-1997, recorded in a special registration system. RESULTS: Consistent with other investigations, injuries from falls made up 25% of all injuries; they were the cause of 28% of all contusions, 32% of all fractures, 61% of all sprains and strains, 40% of all injuries to lower extremities, and 62% of all injuries to the chest. The proportion of fall injuries in different age groups was U-shaped and constitutes around 40% for men both under 20 years and over 50 years of age, and around 20% for those between these ages. Frequent types of injury mechanisms other than falls and slips were: getting caught (22%), contact with objects or persons (28%), foreign body (9%), and cuts (9%). CONCLUSIONS: Use of proportionate data gave a detailed description of injuries from falls and slips, showing important areas for prevention. To avoid a possible misclassification of fall injuries in future studies, it is recommended to include an extra specific variable: whether falling or slipping preceded the crash phase of the injury or not. PMID- 10797508 TI - A comparison of approaches to modeling the relationship between ergonomic exposures and upper extremity disorders. AB - BACKGROUND: For a study of upper extremity musculoskeletal disorders among automobile manufacturing workers, an ergonomic exposure index was constructed by summing ten psychophysical (interview) items. Here we explore the sensitivity of the exposure-response relationship to the formulation of that index. METHODS: Five alternative exposure indices were constructed: three a priori weighting schemes and two sets of weights derived from multivariate regression coefficients. In addition, structural equation modeling was performed with LISREL. RESULTS: The original index and a priori weighting schemes had similar associations with upper extremity disorders, adjusted for nonoccupational covariates. A reasonable model fit was achieved in LISREL after two modifications; the standardized solution showed that nonneutral postures were significantly related to upper extremity signs and symptoms. CONCLUSIONS: In this large population, with adequate range of exposures, the exposure-response relationship appeared generally robust to the mathematical formulation of the exposure index. Among the available exposure variables, postural strain had the strongest association. PMID- 10797509 TI - Years of productivity lost among injured workers in Washington state: modeling disability burden in workers' compensation. AB - BACKGROUND: Few studies have examined the impact of disability from nonfatal work related injuries and illnesses. We developed Years of Productivity Lost (YPL) as a measure of the burden of disability. METHODS: YPL was calculated for incident workers' compensation claims with compensable injuries filed in 1986 in Washington State. Final work disability status was determined in June, 1993. We estimated YPL by adding actual accumulated time loss and future lost productivity predicted from permanent partial disability awards and pensions. RESULTS: Back and neck sprains were associated with the highest YPL followed by sprains of the lower and upper extremities. Using actual accumulated time loss, we calculated 14,624 years of productivity lost for compensable injuries in 1986. After including predicted lost productivity, YPL ranged from 28,017 to 33,502 years. CONCLUSIONS: YPL can be used to quantify the burden of disability due to occupational injuries and illnesses and to prioritize efforts to prevent long term disability. PMID- 10797510 TI - Increase in airway hyperresponsiveness among workers exposed to methylene diphenyldiisocyanate compared to workers exposed to toluene diisocyanate at a petrochemical plant in Korea. AB - BACKGROUND: The purpose of this study was to investigate the prevalence of airway hyperresponsiveness induced by methylene diphenyldiisocyanate (MDI) and toluene diisocyanate (TDI) at a petrochemical industry complex in Korea. METHODS: Questionnaires, allergic skin test, and nonspecific airway hyperresponsiveness (AHR) were studied in 64 exposed workers and 27 control subjects. Questionnaires included questions about symptoms of cough, wheezing, chest tightness, dyspnea, rhinorrhea, sneezing, itching, stuffiness, tearing, urticaria, sore throat, and exacerbating time. Methacholine challenge tests were done. Bronchial responsiveness (BRindex) defined as log (% fall in FEV(1))/log (last concentration of methacholine +10). RESULTS: Prevalence of AHR (PC20 FEV(1) < 16.0 mg/mL of methacholine) was higher in MDI-exposed workers than in TDI-exposed workers [4/20 (20%) vs. 2/42 (4.7%), P<0.05]. Twenty-three workers (36%) of all subjects had respiratory symptoms. MDI-exposed workers, in comparison with control subjects, had higher BRindex (0.73+/-0.04 vs. 0.62+/-0.02, P<0.05). Workers exposed to TDI or MDI who had respiratory symptoms (n = 23), in comparison to workers exposed to TDI or MDI without respiratory symptoms (n = 41), had significantly higher BRindex (0.82+/-0.06 vs. 0.60+/-0.02, P<0.05). FEV(1) was significantly negatively correlated with BRindex (r = -0.253, P<0.05). BRindex was not correlated with atopy, smoking status, and exposure duration. CONCLUSIONS: These findings suggest that workers exposed to MDI are at a higher risk of asthma in comparison with TDI-exposed workers and control subjects at a petrochemical plant in Korea. PMID- 10797511 TI - Researchers should talk to workers. PMID- 10797512 TI - Re: liver damage among shoe repairers. PMID- 10797513 TI - Bootstrap confidence intervals: when, which, what? A practical guide for medical statisticians. AB - Since the early 1980s, a bewildering array of methods for constructing bootstrap confidence intervals have been proposed. In this article, we address the following questions. First, when should bootstrap confidence intervals be used. Secondly, which method should be chosen, and thirdly, how should it be implemented. In order to do this, we review the common algorithms for resampling and methods for constructing bootstrap confidence intervals, together with some less well known ones, highlighting their strengths and weaknesses. We then present a simulation study, a flow chart for choosing an appropriate method and a survival analysis example. PMID- 10797514 TI - Estimating HIV incidence using dates of both HIV and AIDS diagnoses. AB - Knowledge of HIV incidence is important to formulate sensible strategies aimed at controlling the HIV/AIDS epidemic. Back-projection is one of the methods for reconstructing the HIV incidence curve from AIDS incidence data. However, because of the low risk of developing AIDS during the first few years after infection, precise estimates of HIV incidence for the recent past are unlikely if we use AIDS incidence data only. As a result there have been recent attempts to use, not only the date of AIDS diagnosis, but also to use the date of their first positive HIV test. The objective of this paper is to incorporate into back-projection the additional information provided by those individuals who have tested HIV positive but have not yet developed AIDS. This adds information on a very large number of other individuals, and provides the hope that the precision of back-projection is improved considerably. The date of a positive HIV test or an AIDS diagnosis of an individual, whichever comes first, is used in a generalized convolution equation for the purpose of back-projection. The method is illustrated by an application to Australian HIV and AIDS data. Study results show that dramatic improvement in precision is gained for estimates of HIV incidence in recent years when both HIV and AIDS diagnosis dates are used on all individuals. PMID- 10797515 TI - Modelling the reproducibility of acoustic rhinometry. AB - This paper presents an application of a linear model with correlated errors to the assessment of the precision of acoustic rhinometry in clinical medicine. The design of the reproducibility study involved 20 healthy subjects measured by five investigators with three replicate measurements of four variates taken from the right and left nasal cavities, altogether 2400 measurements. The parametric model may be structured to account for random variation between subjects, serial correlation within subjects and measurement error. The model-based reproducibility correlation coefficient was estimated to be fair both for the nasal volume measurements and for the nasal minimal cross-sectional area measurements with an overall average of 0.65. An alternative measure, the within subject coefficient of variation, averaged 15 per cent. The choice between the measures of reproducibility depends on the degree of heterogeneity of the studied populations, the presence of time trend and the scale of measurement of the rhinometric variates. In clinical practice, repeated measurements should always be made to attain a reasonable level of reproducibility in acoustic rhinometry. PMID- 10797517 TI - Estimation of the contact rate in a seasonal SEIR model: application to chickenpox incidence in France. AB - This work was an attempt at analysing the seasonal pattern of varicella epidemics in France by fitting a susceptibles-exposed-infectives-recovered (SEIR) model with a periodic contact rate function to weekly chickenpox incidence data collected over the period 1991 to 1996. The contact rate was assumed to be either a continuous, or a piecewise constant, periodic function. Both assumptions gave a reasonable fit to the data and yielded estimates of the incubation and infectious periods that were consistent with values derived on clinical or serological grounds. Seasonal fluctuation of the estimated contact rate function was likely to reflect the impact of school holidays on chickenpox diffusion among schoolchildren. PMID- 10797516 TI - Choice of scale for cubic smoothing spline models in medical applications. AB - The determination of the functional form of the relationship between an outcome variable and one or more continuous covariates is an important aspect of the modelling of medical data. For correct interpretation of the data it is essential that the functional form be specified at least approximately correctly. I show that for given model complexity, logarithmic transformation of a covariate can greatly improve the fit of one of the most useful and convenient non-parametric regression models, the cubic smoothing spline. A mathematical rationale for the idea is given. I propose a diagnostic for deciding initially whether a log transformation is needed. The method is illustrated using several medical data sets. No special software other than that used for fitting the spline models is needed. PMID- 10797520 TI - Correction PMID- 10797518 TI - Genetic analysis of the age at menopause by using estimating equations and Bayesian random effects models. AB - Multi-wave self-report data on age at menopause in 2182 female twin pairs (1355 monozygotic and 827 dizygotic pairs), were analysed to estimate the genetic, common and unique environmental contribution to variation in age at menopause. Two complementary approaches for analysing correlated time-to-onset twin data are considered: the generalized estimating equations (GEE) method in which one can estimate zygosity-specific dependence simultaneously with regression coefficients that describe the average population response to changing covariates; and a subject-specific Bayesian mixed model in which heterogeneity in regression parameters is explicitly modelled and the different components of variation may be estimated directly. The proportional hazards and Weibull models were utilized, as both produce natural frameworks for estimating relative risks while adjusting for simultaneous effects of other covariates. A simple Markov chain Monte Carlo method for covariate imputation of missing data was used and the actual implementation of the Bayesian model was based on Gibbs sampling using the freeware package BUGS. PMID- 10797521 TI - Correction PMID- 10797519 TI - Analysing the effects of anaemia on local recurrence of head and neck cancer when covariate values are missing. AB - We present a case study in the analysis of the prognostic effects of anaemia and other covariates on the local recurrence of head and neck cancer in patients who have been treated with radiation therapy. Because it is believed that a large fraction of the patients are cured by the therapy, we use a failure time mixture model for the outcomes, which simultaneously models both the relationship of the covariates to cure and the relationship of the covariates to local recurrence times for subjects who are not cured. A problematic feature of the data is that two covariates of interest having missing values, so that only 75 per cent of the subjects have complete data. We handle the missing-data problem by jointly modelling the covariates and the outcomes, and then fitting the model to all of the data, including the incomplete cases. We compare our approach to two traditional methods for handling missingness, that is, complete-case analysis and the use of an indicator variable for missingness. The comparison with complete case analysis demonstrates gains in efficiency for joint modelling as well as sensitivity of some results to the method used to handle missing data. The use of an indicator variable yields results that are very similar to those from joint modelling for our data. We also compare the results obtained for the mixture model with results obtained for a standard (non-mixture) survival model. It is seen that the mixture model separates out effects in a way that is not possible with a standard survival model. In particular, conditional on other covariates, we find strong evidence of an association between anaemia and cure, whereas the evidence of an association between anaemia and time to local recurrence for patients who are not cured is weaker. PMID- 10797522 TI - Correction PMID- 10797523 TI - Comparative calibration without a gold standard Y. Lu, K. Ye, A. Mathur, S. Hul, T. Fuerst and H. Genant, statistics in medicine 1997; 16:1889-1905 PMID- 10797524 TI - Re: Comparative calibration without a gold standard. Y. Lu, K. Ye, A. Mathur, S. Hul, T. Fuerst and H. Genant, Statistics in Medicine 1997; 16:1889-1905. PMID- 10797525 TI - Genetic abnormalities in marginal zone B-cell lymphoma. AB - Marginal zone B-cell lymphoma (MZBCL) including extranodal mucosa-associated lymphoid tissue (MALT)-type lymphoma, nodal, and splenic MZBCL represents a distinct subtype of B-non-Hodgkin's lymphoma. Recently, important progress in the elucidation of the genetic mechanisms underlying the pathogenesis and disease progression of these lymphomas has been made. The API2 gene, an inhibitor of apoptosis, and the novel MLT gene have been found to be altered by the t(11;18)(q21;21), which represents the most frequent structural chromosomal abnormality in extranodal low-grade MALT lymphoma. Another gene involved in the regulation of apoptosis, the BCL10 gene, has been cloned from a MALT lymphoma cytogenetically characterized by the t(1;14)(p22;q32). Along the same lines, inactivating mutations of the proapoptotic FAS gene have been detected in a relatively high proportion of extranodal MZBCLs. Considering these data and the fact that at least some MALT lymphomas show low levels of apoptosis and seem to escape from FAS-mediated apoptosis one may speculate that abrogation of apoptosis constitutes a central pathogenetic mechanism in the development of these lymphomas. The pathogenetic role of trisomy 3, the most frequent numerical chromosomal change of MZBCL, is not known. The minimal overrepresented region has been delineated to 3q21-23 and 3q25-29 using comparative genomic hybridization. The BCL6 proto-oncogene, located on 3q27, which is rearranged in some MZBCL and a high proportion of large cell B-cell lymphomas with extranodal localization, represents one of the candidate genes residing in these critical regions. PMID- 10797528 TI - Recent publications in hematological oncology. PMID- 10797526 TI - Association of gastric and Waldeyer's ring lymphoma: a molecular study. AB - The coexistence of Waldeyer's ring and gastrointestinal non-Hodgkin's lymphomas at presentation is well known. Moreover, localized gastrointestinal relapses following successful treatment of lymphomas of Waldeyer's ring and thyroid lymphomas occurring after a prolonged disease-free interval have also been described. We report two cases of concomitant lymphoma in Waldeyer's ring and stomach. On the basis of the molecular analysis of the immunoglobulin heavy chain gene rearrangements, two different patterns of concomitant involvement by a lymphoma in Waldeyer's ring and in the gastrointestinal region seem to exist. One is represented by the preferential dissemination of the lymphoma from one site to the other, the second by the apparently independent growth of clonally unrelated lymphomas at each site. PMID- 10797527 TI - Primary central nervous system lymphoma: a clinicopathological study of 28 cases. AB - A group of 28 consecutive patients (mean age 59 years) with primary central nervous system lymphoma (PCNSL) was treated with different regimens, including steroids only, radiotherapy (RT), chemotherapy or combinations of all. Lymphoma was classified as high grade malignant B-cell non-Hodgkin's lymphoma of the diffuse large cell type in each of these cases. RT alone led to tumour remission in more than 70 per cent, survival could be prolonged with additional chemotherapy. Thirteen patients were treated with chemotherapy alone; nine of them received a novel combined intraventricular and systemic polychemotherapy protocol based on high dose methotrexate (MTX) and high dose cytarabine (ara-C). The response rate was 90 per cent with 80 per cent complete responses. Neurotoxicity, i.e. white matter lesions associated with severe cognitive dysfunction affected both patients surviving RT more than a year and patients treated with combination RT/chemotherapy. Confluent white matter hyperintense lesions were detectable on MRI in three out of 13 patients treated with chemotherapy alone, however, cognitive dysfunction has not been detected in these patients. PMID- 10797529 TI - A neuroscientist looks at philosophy: response to Beedle (1999). PMID- 10797530 TI - Developmental expression of OSP/claudin-11. AB - Oligodendrocyte-specific protein (OSP/claudin-11) is a major component of CNS myelin and has been recently added to the claudin family of tight junction proteins. In this study, the developmental expression of OSP/claudin-11 was determined using in situ hybridization, immunohistochemistry (IH), and Western blot analysis. OSP/claudin-11 mRNA was expressed in a bimodal fashion. During prenatal development, OSP/claudin-11 mRNA was abundant in developing meninges, in areas adjacent to cartilage, and in mesoderm. In postnatal animals, OSP/claudin 11 was expressed primarily in developing oligodendrocytes and to a lesser extent, in testes. Double-labeled IH using O2-A progenitor cells revealed that OSP/claudin-11 expression occurs from the early progenitor stage and continues in mature oligodendrocytes. Electron microscopic IH localized OSP/claudin-11 to laminar myelin in the adult CNS. Western blot analysis of OSP/claudin-11 in developing brain revealed the expression of two separate transcripts that were developmentally regulated. These data demonstrate that OSP/claudin-11 expression is highly regulated during development and, therefore, may play an important role in growth and differentiation of oligodendrocytes and other cells outside the CNS. PMID- 10797531 TI - Laminin modulates neuritogenesis of developing rat retinal ganglion cells through a protein kinase C-dependent pathway. AB - Dissociated cells from rat retinae (P2-P21) were cultured to investigate interactions between brain-derived neurotrophic factor (BDNF), various substrates (poly-L-lysine, collagen, and laminin), and protein kinases upon the neuritogenesis of retinal ganglion cells (RGCs). We found that BDNF-promoted neuritogenesis was enhanced by forskolin in RGCs from rats at P2-P21 plated on either poly-L-lysine or collagen. In contrast, in cultures with a laminin substrate, the enhancer effect of forskolin was observed only in RGCs taken from the retina of rats at P2-P6. Laminin blocked the enhancement of BDNF-induced RGCs neuritogenesis by forskolin, in RGCs from either P14 or P21, and induced a tenfold increase of protein kinase C (PKC) activity compared to poly-L-lysine. This blockade was reverted with a selective PKC inhibitor and was reproduced in poly-L-lysine cultures of P14-P21 RGCs with a PKC activator. Because axotomized RGCs need both BDNF and forskolin to regenerate, we suggest that laminin can hinder this effect by simultaneous PKC activation according to a developmentally regulated pattern. We further propose a model of interaction in the optic pathways triggered by BDNF, forskolin, and laminin that may be useful in elucidating some of the biological effects seen with regenerating axons. PMID- 10797532 TI - Conditionally-immortalized astrocytic cell line expresses GAD and secretes GABA under tetracycline regulation. AB - We have engineered conditionally-immortalized mouse astrocytes to express beta galactosidase or GAD(65) in a tetracycline-controlled fashion. The engineered cell lines, BASlinbetagal and BASlin65, divide at 33 degrees C but cease division at 39 degrees C. We carried out morphological and biochemical analyses to further understand GABA production and release, and to determine the suitability of these cells for transplantation. Using the BASlinbetagal cell line, we showed a dramatic regulation of beta-galactosidase expression by tetracycline. The BASlin65 cell line showed functional GAD(65) enzymatic activity and GABA production, both of which were suppressed by growth in the presence of tetracycline. When cultured in the absence of tetracycline, BASlin65 cells have a total GABA content equal to or greater than other GABA-ergic cell lines. Immunofluorescence microscopy revealed that GAD(65) had a distinct perinuclear localization and punctate staining pattern. GABA, on the other hand, showed diffuse staining throughout the cytoplasm. BASlin65 cells not only synthesize GABA, they also release it into the extracellular environment. Their ability to produce and release significant amounts of GABA in a tetracycline-regulated manner makes BASlin65 cells a useful cellular model for the study of GABA production and release. Furthermore, their non-tumorigenicity makes them excellent candidates for transplantation into specific regions of the brain to provide a localized and regulatable source of GABA to the local neuronal circuitry. PMID- 10797533 TI - Functional implications of the noradrenergic-cholinergic switch induced by retinoic acid in NB69 neuroblastoma cells. AB - Some neuroblastoma cell lines change their neurotransmitter phenotype from noradrenergic to cholinergic under retinoic acid treatment. Such "neurotransmitter switch" seems to be a consequence of changes in the expression and activity of the biosynthetic machinery for both neurotransmitters. In this study, we have characterized this "neurotransmitter switch" induced by retinoic acid in a human neuroblastoma cell line (NB69) showing catecholaminergic characteristics. Retinoic acid treatment reduced tyrosine hydroxylase activity and noradrenaline levels in NB69 cells but did not modify the expression of this enzyme. Moreover, the calcium-dependent release of [(3)H]noradrenaline in control cells was highly reduced by retinoic acid treatment. On the other hand, NB69 cells treated with retinoic acid enhanced the expression of choline acetyltransferase and acquired the capability to release [(3)H]acetylcholine in a calcium-dependent way. In addition, we found that the expression of the vesicular monoamine transporter 2 (VMAT2) and the vesicular acetylcholine transporter (VAChT) was increased in those cells treated with retinoic acid. Immunostaining revealed that retinoic acid treatment changed the cellular distribution of both vesicular monoamine transporter 2 and vesicular acetylcholine transporter. In conclusion, retinoic acid induces a noradrenergic to cholinergic switch in NB69 cells by acting at several levels of the neurotransmitter phenotypic expression. PMID- 10797534 TI - Phosphatidylinositol 3-kinase mediates neuroprotection by estrogen in cultured cortical neurons. AB - It has been shown that estrogen replacement in menopausal women is effective in slowing down the progression of cognitive impairment in Alzheimer's disease. Although recent studies have demonstrated the neuroprotective effects of estrogen, the precise mechanism of neuroprotection has not been elucidated. In the present study, we show that the phosphatidylinositol 3-kinase (PI3-K) cascade is involved in the neuroprotective mechanism stimulated by estrogen. Exposure to glutamate reduced the viability of rat primary cortical neurons. Pretreatment with 10 nM 17beta-estradiol significantly attenuated the glutamate-induced toxicity. This neuroprotective effect of 17beta-estradiol was blocked by co administration with LY294002, a selective PI3-K inhibitor, but not by co administration with PD98059, a selective mitogen activated protein kinase kinase inhibitor. Pretreatment with ICI182780, a specific estrogen receptor antagonist, also blocked the neuroprotection. Immunoblotting assay revealed that treatment with 17beta-estradiol induced the phosphorylation of Akt/PKB, an effector immediately downstream of PI3-K. These results suggest that PI3-K mediates the neuroprotective effect of 17beta-estradiol against glutamate-induced neurotoxicity. PMID- 10797535 TI - Photoreceptor phagocytosis selectively activates PPARgamma expression in retinal pigment epithelial cells. AB - Phagocytosis of tips of rod outer segments (ROS) by retinal pigment epithelial (RPE) cells is vitally important for maintaining structural and functional integrity of the retina. We previously reported that receptor-mediated specific phagocytosis of ROS induces expression of early response genes coding for transcription factors. Here we study the expression of peroxisome proliferator activated receptors (PPAR) -alpha, -delta (beta) and -gamma during ROS phagocytosis of rat RPE cells in primary cell culture, using competitive quantitative RT-PCR. During phagocytosis of ROS (but not of latex particles) by RPE cells, RT-PCR revealed a transient increase in PPARgamma mRNA expression, that peaked at 4-6 hr. We sequenced and described two alternatively spliced variants of rat PPARgamma: rPPARgamma1a and rPPARgamma1b. Both of these, along with the recently described rPPARgamma2 were induced by ROS phagocytosis. PPARalpha and PPARdelta mRNA expression was also detected in RPE cells, but the level of expression did not change during ROS phagocytosis. All-trans-retinoic acid and prostaglandin E(2) (PGE(2)) selectively potentiated both basal and ROS phagocytosis-induced PPARgamma expression. All-trans-retinoic acid had the opposite inhibitory effect on PPARalpha and PPARdelta expression. Cycloheximide had a dual action on PPARgamma expression in RPE cells: it enhanced expression under basal conditions but repressed expression induced by ROS phagocytosis. It also stimulated expression of PPARalpha but had no effect on PPARdelta. Selective activation of PPARgamma may play an important role in regulating the expression of target genes that are involved in lipid and fatty acid metabolism in the photoreceptor renewal process. PMID- 10797536 TI - Synthesis and degradation of cytochrome oxidase subunit mRNAs in neurons: differential bigenomic regulation by neuronal activity. AB - Cytochrome oxidase (CO) plays a key role in oxidative capacity of neurons and serves as a sensitive indicator of neuronal activity. The mechanism(s) involved in the regulation of this bigenomic-encoded mitochondrial enzyme is still not clearly understood. Previous studies have shown changes in the level of its subunit mRNAs encoded by the nuclear or mitochondrial genome in response to changing neuronal activity. Our goals in the present study were to determine whether such changes were due to RNA synthesis rate or stability or both. The level of CO activity of neurons in primary cultures assayed histochemically was increased after depolarizing KCl treatment. The steady-state levels of CO subunit II (CO II; mitochondrial-encoded) and IV (CO IV; nuclear-encoded) mRNAs were up regulated in response to 5 hr of 20 mM KCl treatment. By using gene-specific probes, the relative rates of synthesis of CO II and IV mRNA were elevated significantly after KCl treatment (P < 0.05). The degradation of CO II and IV mRNAs was monitored by (3)H-uridine pulse-chase labeling, which revealed half lives of 84 min for CO II mRNA and 50 min for CO IV mRNA. Under KCl treatment, the half-life of CO IV was increased to 102 min, but there was no statistically significant change in the half-life of CO II mRNA. These results indicate that mitochondrial subunit CO II mRNA is regulated mainly at the transcriptional level, whereas the nuclear subunit CO IV mRNA is regulated at both the synthetic and the degradative levels. Both subunits, however, are tightly governed by neuronal activity. PMID- 10797537 TI - Effect of ethanol on muscarinic receptor-induced calcium responses in astroglia. AB - The effects of ethanol on muscarinic receptor-mediated calcium responses were investigated in individual primary rat astrocytes and human 132 1N1 astrocytoma cells using indo-1/AM and image cytometry. After a 30-min incubation, carbachol induced calcium responses were inhibited only at 100 or 250 mM ethanol. The effects of ethanol were more pronounced and occurred at lower concentrations with longer exposures, with significant inhibition seen at 10 mM following a 24-hr incubation. Thapsigargin- and glutamate-induced responses were unaffected by ethanol, indicating some selectivity in this inhibition. Upon removal of ethanol, inhibition of calcium responses persisted for up to 6-12 hr, with carbachol responses returning to control levels by 24 hr after washout. Ethanol exposure did not affect muscarinic-receptor binding in astrocytoma cells, but inhibited carbachol-induced IP(3) formation. Inhibition of (3)H-thymidine incorporation by ethanol also persisted upon removal of the alcohol, with a time-dependency similar to that of the calcium responses. These results indicate that ethanol inhibits muscarinic receptor-induced calcium responses in astroglia in a concentration- and duration-dependent manner. They also show that co-incubation with ethanol is not necessary for this effect, suggesting that long-term exposure to ethanol may modify, in a reversible manner, the coupling of muscarinic receptors with its effector. This effect of ethanol may play a role in ethanol's inhibition of carbachol-induced thymidine incorporation. PMID- 10797538 TI - Vasoactive amine responses in murine cerebrovascular endothelial cells as measured by extracellular acidification rates. AB - The Cytosensortrade mark microphysiometer device is capable detecting cellular responses to specific bioactive ligands by measuring the extracellular acidification rate (ECAR). Using microphysiometry, we were able to determine that cerebovascular endothelial cells derived from SJL/J mice were more sensitive to serotonin (maximal ECAR response at 100 nM), whereas BALB/c-derived cerebrovascular endothelial cells (CVE) were relatively insensitive (maximal ECAR response at 30 microM). Serotonin (5HT)(1) and 5HT(2) receptor antagonists inhibited the serotonin-mediated increases in ECAR. The cells' responses to histamine in both strains were similar (maximal ECAR required 100 microM of histamine). H(1) and H(3) receptor subtype antagonists specifically inhibited the histamine responses. Bradykinin also revealed sensitivity differences in that maximal ECAR changes for CVE from SJL/J mice could be observed with 1 microM, as compared to the ECAR responses from BALB/c mice CVE, which required 10 microM. Exposure to bradykinin antagonists revealed that the response was due to the stimulation of B(2) receptors. These microphysiometry results reveal that the cerebrovascular endothelial cells of SJL/J mice tend to be more sensitive to vasoactive substances than those of BALB/c mice. PMID- 10797540 TI - Presynaptic excitability changes following traumatic brain injury in the rat. AB - Pathological processes affecting presynaptic terminals may contribute to morbidity following traumatic brain injury (TBI). Posttraumatic widespread neuronal depolarization and elevated extracellular potassium and glutamate are predicted to alter the transduction of action potentials in terminals into reliable synaptic transmission and postsynaptic excitation. Evoked responses to orthodromic single- and paired-pulse stimulation were examined in the CA1 dendritic region of hippocampal slices removed from adult rats following fluid percussion TBI. The mean duration of the extracellularly recorded presynaptic volley (PV) increased from 1.08 msec in controls to 1.54 msec in slices prepared at 1 hr postinjury. There was a time-dependent recovery of this injury effect, and PV durations at 2 and 7 days postinjury were not different from controls. In slices removed at 1 hr postinjury, the initial slopes of field excitatory postsynaptic potentials (fEPSPs) were reduced to 36% of control values, and input/output plots revealed posttraumatic deficits in the transfer of excitation from pre- to postsynaptic elements. Manipulating potassium currents with 1.0 mM tetraethylammonium or elevating potassium ion concentration to 7.5 mM altered evoked responses but did not replicate the injury effects to PV duration. Paired pulse facilitation of fEPSP slopes was significantly elevated at all postinjury survivals: 1 hr, 2 days, and 7 days. These results suggest two pathological processes with differing time courses: 1) a transient impairment of presynaptic terminal functioning affecting PV durations and the transduction of afferent activity in the terminals to reliable synaptic excitation and 2) a more protracted deficit to the plasticity mechanisms underlying paired-pulse facilitation. PMID- 10797539 TI - Regulation of the neurotensin NT(1) receptor in the developing rat brain following chronic treatment with the antagonist SR 48692. AB - The aim of the present study was to investigate the role of neurotensin in the regulation of NT(1) receptors during postnatal development in the rat brain. Characterization of the ontogeny of neurotensin concentration and [(125)I]neurotensin binding to NT(1) receptors in the brain at different embryonic and postnatal stages showed that neurotensin was highly expressed at birth, reaching peak levels at postnatal day 5 (P5) and decreasing thereafter. The transient rise in neurotensin levels preceded the maximal expression of NT(1) receptors, observed at P10, suggesting that neurotensin may influence the developmental profile of NT(1) receptors. Using primary cultures of cerebral cortex neurons from fetal rats, we showed that exposure to the neurotensin agonist JMV 449 (1 nM) decreased (-43%) the amount of NT(1) receptor mRNA measured by reverse transcription-PCR, an effect that was abolished by the nonpeptide NT(1) receptor antagonist SR 48692 (1 microM). However, daily injection of SR 48692 to rat pups from birth for 5, 9, or 15 days did not modify [(125)I]neurotensin binding in brain membrane homogenates. Moreover, postnatal blockade of neurotensin transmission did not alter the density and distribution of NT(1) receptors assessed by quantitative autoradiography nor NT(1) receptor mRNA expression measured by in situ hybridization in the cerebral cortex, caudate putamen, and midbrain. These results suggest that although NT(1) receptor expression can be regulated in vitro by the agonist at an early developmental stage, neurotensin is not a major factor in the establishment of the ontogenetic pattern of NT receptors in the rat brain. PMID- 10797541 TI - Missense point mutations of tau to segregate with FTDP-17 exhibit site-specific effects on microtubule structure in COS cells: a novel action of R406W mutation. AB - Missense and splicing point mutations have been found in the tau gene in families with frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17). Of these mutations, we examined four exonic missense point mutations (G272V, P301L, V337M and R406W) in 3-repeat or 4-repeat tau isoform on the transfection experiment. The effects of two mutations (G272V or P301L) on microtubules were subtle whereas those of two other mutations (V337M or R406W) were dramatically significant when these two mutations were constructed into 3-repeat tau but not into 4-repeat tau. The R406W mutation induced an alternation of microtubules to form dotted or fragmented forms retaining colocalization of tau with tubulin whereas the V337M mutation predominantly disrupted microtubule networks and diminished colocalization of tau and tubulin. The effect of the mutations on microtubules were thus site-dependent and isoform-dependent. Tau with R406W mutation was found to be colocalized with tubulin without filamentous structures on confocal views, suggesting that the carboxyl region of tau played a different role from tubulin-binding domain on microtubule assemble. Another abnormal property was identified in tau with R406W mutation that failed to suffer phosphorylation. Thus, diverse effects of tau mutations on microtubules may explain the various clinicopathologies of FTDP-17 and related tauopathies. PMID- 10797542 TI - Apoptosis induced by 2-chloro-adenosine and 2-chloro-2'-deoxy-adenosine in a human astrocytoma cell line: differential mechanisms and possible clinical relevance. AB - We have previously demonstrated that 2-chloro-adenosine (2-CA) can induce apoptosis of rat astroglial cells (Abbracchio et al. [1995] Biochem. Biophys. Res. Commun. 213:908-915). In the present study, we have characterized, for the first time, the effects induced on a human astrocytoma cell line (ADF cells) by both 2-CA and its related analog 2-chloro-2'-deoxy-adenosine (2-CdA, that is employed as anti-cancer agent in chronic lymphoid malignancies). Exposure of these cells to either adenosine analog resulted in time- and concentration dependent apoptosis. Experiments with pharmacological agents known to interfere with adenosine receptors, its membrane transporter, and intracellular nucleoside kinases showed that: (i) cell death induced by either adenosine analog did not depend on extracellular adenosine receptors, but on a direct intracellular action; however, only in the case of 2-CA, was entry into cells mediated by the specific nitrobenzyl-tioinosine-sensitive transporter; (ii) for both adenosine analogs, induction of apoptosis required the phosphorylation/activation by specific intracellular nucleoside kinases, i.e., adenosine kinase for 2-CA, and deoxycytidine kinase for 2-CdA. In addition, only in the case of 2-CdA, was induction of apoptosis preceded by a block of cells at the G2/M phase of the cell cycle. Finally, at concentrations of either analog that killed about 80-90% of astrocytoma cells, a significantly lower effect on the viability of primary cortical neurons was observed. In conclusion, both adenosine analogs can trigger apoptosis of human astrocytoma cells, albeit with different mechanisms. This effect together with the relative sparing of neuronal cells, may have potential clinical implications for the therapy of tumors of glial origin. PMID- 10797544 TI - GFAP-positive and myelin marker-positive glia in normal and pathologic environments. AB - The data herein demonstrate that in addition to the well-characterized myelin marker-positive, glial fibrillary acidic protein (GFAP)-negative, membrane sheet bearing oligodendrocytes, another type of myelin marker-positive, process-bearing glia exists in normal and pathologic conditions. This second type of myelin marker-positive glia expresses GFAP, and therefore these cells have been referred to as mixed phenotype glia. Although mixed phenotype glia have been documented previously, their identity and function have remained a mystery. The goal of this immunocytochemical study was to further characterize these cells. Using the MBPlacZ transgenic mouse in which beta-galactosidase is under the control of the myelin basic protein (MBP) gene promoter, GFAP-positive/beta-galactosidase positive and myelin/oligodendrocyte-specific protein (MOSP)-positive/beta galactosidase-positive cells were detected in subcortical white matter and in perivascular locations within cerebral white and gray matter. In cultures prepared from highly enriched myelin marker-positive immature glia, mixed phenotype glia were detected that were GFAP-positive and either MOSP-, MBP-, O1-, and O4-positive. The expression of multiple myelin markers by mixed phenotype glia may suggest that these cells are of oligodendrocyte origin. Increased numbers of MOSP-positive/GFAP-positive mixed phenotype glia were detected in sections from adult hypomyelinated brain from shiverer, quaking, and PKU mice compared to myelinated control adult mouse brain. Similarly, cultures from control brain exposed to elevated pH for 2-3 weeks showed dramatically increased numbers of mixed phenotype glia (80%) compared to control (<10%). Increased numbers of mixed phenotype glia also were detected in shiverer cultures (40%). Since increases in the number of mixed phenotype glia occur in shiverer, quaking, and PKU mouse brain, these data suggest that mixed phenotype glia contribute to gliosis in pathologic white matter. PMID- 10797543 TI - Expression of alpha(2)-macroglobulin receptor/low density lipoprotein receptor related protein (LRP) in rat microglial cells. AB - Low density lipoprotein receptor-related protein (LRP) participates in the uptake and degradation of several ligands implicated in neuronal pathophysiology including apolipoprotein E (apoE), activated alpha(2) -macroglobulin (alpha(2)M*) and beta-amyloid precursor protein (APP). The receptor is expressed in a variety of tissues. In the brain LRP is present in pyramidal-type neurons in cortical and hippocampal regions and in astrocytes that are activated as a result of injury or neoplasmic transformation. As LRP is expressed in the monocyte/macrophage cell system, we were interested in examining whether LRP is expressed in microglia. We isolated glial cells from the brain of neonatal rats and LRP was immunodetected both in microglial cells and in astrocytes expressing glial fibrillar acidic protein (GFAP). Microglial cells were able to bind and internalize LRP-specific ligand, alpha(2)M*. The internalization was inhibitable by RAP, with a Kd of 1.7 nM. The expression of LRP was up-regulated by dexamethasone, and down-regulated by lipopolysaccharide (LPS), gamma interferon (IFN-gamma) or a combination of both. LRP was less sensitive to dexamethasone in activated astrocytes than in microglia. We provided the first analysis of LRP expression and regulation in microglia. Our results open the possibility that microglial cells could be related to the participation of LRP and its ligands in different pathophysiological states in brain. PMID- 10797545 TI - Ephrins stimulate or inhibit neurite outgrowth and survival as a function of neuronal cell type. AB - The Eph family of tyrosine kinase receptors and ligands play key roles in cell segregation and axon targeting in the developing nervous system. Interactions between the ligands and receptors cause repulsion or degeneration of receptor positive axons from several brain regions including the retina, hippocampus, thalamus, and midbrain dopaminergic system. We extend these previous observations by showing that three A-ephrins also negatively regulate the growth of neurites from striatal and olfactory neurons. In addition to negative effects, however, we also report a trophic activity of the A-ephrins: Ephrin-A2 and A5 promote survival and neurite outgrowth of sympathetic neurons. These observations provide support to the notion that ephrins may function as either negative or positive signals in the developing nervous system. PMID- 10797546 TI - Redistribution of cholesterol in oligodendrocyte membrane sheets after activation of distinct signal transduction pathways. AB - Cultured oligodendrocytes produce extensive membrane sheets that contain an internal lacy network of vein-like structures composed of microtubules, actin filaments, and 2'3'-cyclic nucleotide 3'-phosphohydrolase (CNPase). These cytoplasmic vein-like structures surround domains of myelin basic protein (MBP). Using the antibiotic filipin, that binds to cholesterol, the relationship between plasma membrane cholesterol and cytoskeleton in membrane sheets was examined. Our results show that cholesterol was relatively uniformly distributed within the plasma membranes of prefixed control oligodendrocyte membrane sheets. When live cultures were extracted with Triton X-100, however, a subpopulation of cholesterol molecules remained colocalized with cytoskeleton in the membrane sheets. Activation of two well-characterized signaling pathways that differentially affect microtubule and actin filament stability in membrane sheets resulted in an apparent massive lateral movement of cholesterol molecules away from membrane regions overlying internal MBP domains to membrane tracts directly overlying cytoplasmic cytoskeletal veins. Depolymerization of microtubules by colchicine resulted in redistribution of cholesterol directly over actin filaments, whereas depolymerization of actin filaments by cytochalasin B resulted in redistribution of cholesterol directly over CNPase/microtubular veins. These data suggest that cholesterol forms an association with cytoskeletal components or proteins associated with cytoskeleton. These data also suggest that cholesterol, via interactions with cytoskeleton, plays a role in signaling pathways in oligodendrocyte membrane sheets. PMID- 10797547 TI - Role of p53 in DNA strand break-induced apoptosis in organotypic slice culture from the mouse cerebellum. AB - Apoptosis occurs not only in mitotic cells but also in postmitotic neuronal cells. We previously suggested that the tumor suppressor gene p53 is required for DNA strand break-induced apoptosis in dissociated culture of cerebellar granule neurons. In this study, we examined the role of p53 in apoptosis using organotypic slice culture of cerebellum from p53 null and wild-type mice. Exposure to bleomycin significantly increased the numbers of TUNEL-, p53-, and c Jun-positive neurons in the wild-type mouse cerebellar internal granular layer (IGL) and Purkinje cell layer (PL). However, in p53-deficient mice, these responses were not observed. These results are consistent with our previous observations in dissociated neuronal culture showing that the amount of c-Jun protein increases significantly after addition of bleomycin in p53 wild-type cerebellar granule cells. The results presented here also indicate that p53 is involved in DNA strand break-induced apoptosis of fully postmitotic central nervous system neurons and suggest that c-Jun expression occurs downstream of p53 expression. PMID- 10797549 TI - Localization of Bcl-xbeta in the developing and adult rat central nervous system. AB - bcl-xbeta is a novel apoptosis-regulating member of the bcl-x family that has recently been isolated from rats and mice. To explore the functional role of Bcl xbeta, we raised a monoclonal antibody against rat Bcl-xbeta protein and investigated the cellular localization of the molecule in the rat CNS. Immunohistochemistry revealed that, in the fetal and neonatal stages, Bcl-xbeta was intensively and widely expressed in the CNS. Many neurons in the diencephalon and brain stem showed intense cytoplasmic labeling. The immunoreactivity decreased during the postnatal development and reached to the level of adulthood by P14. In the adult brain and spinal cord, labeling was restricted to specific types of neurons and distributed throughout their somata and dendrites. Weak immunoreactivity was present in many CNS regions such as the cerebral cortex, hippocampal dentate gyrus, caudate-putamen, globus pallidus, thalamus, locus ceruleus, pontine nuclei, inferior olive, reticular formation, cerebellar cortex and spinal anterior horn. Amygdaloid nuclei and hippocampal CA1 to CA3 sectors showed restricted expression of Bcl-xbeta in a subset of neurons. Neuronal labeling was almost undetectable in several regions, including the piriform cortex, hypothalamus, posterior column nuclei and spinal posterior horn. These results suggest that Bcl-xbeta plays an important role throughout the CNS in developing stage and may regulate the apoptosis of postnatal CNS neurons. PMID- 10797548 TI - Recruitment of the Arp2/3 complex and mena for the stimulation of actin polymerization in growth cones by nerve growth factor. AB - The growth of axons and dendrites during development and regeneration is regulated by cues in the environment. Many of these cues regulate the actin cytoskeleton of the protrusive structures (like filopodia) of the growth cone that are essential for detecting and responding to cues. Nerve growth factor, which promotes the formation of protrusive structures, stimulated actin polymerization in rat sympathetic growth cones, resulting within 1-2 min in accumulations of F-actin at the distal edge and in splotches of F-actin farther back. We examined the potential involvement of a protein machinery important in at least certain types of actin polymerization in non-neuronal cells. Members of the Arp2/3 complex, p34-Arc and p21-Arc, heavily concentrated in the early accumulations of F-actin, as did one member of the Ena/VASP family (Mena) but not another (VASP). Retention of Arc proteins at preferred sites of actin polymerization did not require polymerization itself. Growth cones of differentiated PC12 cells were similar to sympathetic growth cones in their response to NGF. Introduction into these cells of a peptide that should block the binding of Ena/VASP family proteins to the protein complex at sites of actin polymerization reduced the formation of splotches and filopodia in response to NGF. These results point to the early involvement of the Arp2/3 complex and the Ena/VASP family in growth factor-stimulated actin polymerization that gives rise to protrusive structures at the growth cone. PMID- 10797550 TI - Interleukin-1beta immunoreactivity in identified neurons of the rat magnocellular neurosecretory system: evidence for activity-dependent release. AB - Interleukin-1beta has been demonstrated in neurons of the rat hypothalamus, including cells of the magnocellular neurosecretory system and tuberoinfundibular system (Lechan et al., [1990] Brain Res. 514:135-140). Despite its potential importance to regulation of neuroendocrine function, however, neither the specific cell types that express interleukin-1beta or the conditions that may result in its release have yet been described. Therefore, we utilized a combination of immunocytochemical and immunoelectron microscopic localization, in conjunction with Western blot analysis, on normonatremic, hypernatremic, and lactating rats to assess the site of synthesis and potential secretion characteristics of interleukin-1beta in the rat magnocellular neurosecretory system. Interleukin-1beta immunoreactivity was localized within both oxytocin and vasopressin neurons in the paraventricular, supraoptic, accessory and periventricular hypothalamic nuclei. Additionally, interleukin-1beta immunoreactive fibers were localized in the zona interna and zona externa of the median eminence and in the neurohypophysis. Immunoelectron microscopic analysis revealed that interleukin-1beta immunoreactivity is associated with small spherical structures, distinct from neurosecretory granules, in neurosecretory axons within the neurohypophysis. Furthermore, stimulation of heightened neurosecretory activity via chronic osmotic challenge and lactation resulted in a marked diminution in levels of interleukin-1beta immunoreactivity in the neurohypophysis with a subsequent return to normal levels after cessation of the stimuli. Western blot analysis confirmed the existence of interleukin-1beta protein in the neurohypophysis and provided further evidence for reduction in levels of IL-1beta immunoreactivity after stimulation of secretory activity. These results suggest an endogenous neuronal source of interleukin-1beta exists within the rat magnocellular neurosecretory system under normal physiological conditions. The potential for activity-dependent release of IL-1beta and implications for the involvement of interleukin-1beta in regulation of neurosecretory activity are discussed. PMID- 10797551 TI - Polyglutamine-induced ion channels: a possible mechanism for the neurotoxicity of Huntington and other CAG repeat diseases. AB - CAG repeats resulting in long polyglutamine tracts have been implicated in the pathogenesis of at least eight neurodegenerative diseases including Huntington. Expression of polyglutamine repeats is required for disease and increasing length of the repeats leads to earlier onset of illness (anticipation). Expression of polyglutamine repeats in cultured neurons leads to deposition of intracellular aggregates resembling those found in amyloid diseases, and to neurotoxicity. We report here that polyglutamine can induce large (19-220 pS), long-lived, (lifetime = 6 sec), non-selective (P(cation) = P(anion)) ion channels in planar phospholipid bilayer membranes, and that channel formation is enhanced by acidic pH. We propose that channel formation may be a mechanism of cellular toxicity in Huntington and other CAG repeat disease. PMID- 10797552 TI - Pressure related apoptosis in neuronal cell lines. AB - Pressure is a crucial component of the cellular environment, and can lead to pathology if it varies beyond its normal range. The increased intra-ocular pressures in acute glaucoma are associated with the loss of neurons by apoptosis. Little is known regarding the interaction between pressure and apoptosis at the level of the cell. The model developed in this study examines the effects of elevated ambient hydrostatic pressure directly upon cultured neuronal lines. Conditions were selected to be within physiological limits: 100 mmHg over and above atmospheric pressure for a period of 2 hr, as seen clinically in acute glaucoma. This system can be used to investigate pressure relatively independently of other variables. Neuronal cell line cultures (B35 and PC12) were subjected to pressure conditions in specially designed pressure chambers. Controls were treated identically, except for the application of pressure, and positive controls were treated with a known apoptotic stimulus. Apoptosis was detected by cell morphology changes and by 2 specific apoptotic markers: TUNEL (Terminal transferase dUTP Nick-End Labeling) and Annexin V. These fluorescent markers were detected and quantified by automated Laser Scanning Cytometry. All techniques showed that increased pressure was associated with a greater level of apoptosis compared to equivalent controls. Our results suggest that pressure alone may act as a stimulus for apoptosis in neuronal cell cultures. This raises the possibility of a more direct relationship at the cellular level between pressure and neuronal loss. PMID- 10797553 TI - Heterogeneity of GABA(A) receptor-mediated responses in the human IMR-32 neuroblastoma cell line. AB - The gamma-aminobutyric acid (GABA) response profiles of IMR-32 human neuroblastoma cells were examined using whole-cell patch clamp and RT-PCR techniques. GABA activated a concentration-dependent and bicuculline-sensitive current, and RT-PCR revealed the expression of multiple GABA(A) receptor subunit mRNAs (alpha(1), alpha(3), alpha(4), beta(1), beta(3), gamma(2), and delta). A pharmacological profile of the GABA-induced current was derived using several subunit-selective agents. Diazepam, which requires the presence of a gamma subunit in order to modulate GABA(A) receptor-mediated responses, potentiated GABA-induced currents in a subset of IMR-32 cells. Two populations of GABA activated currents were also evident based on sensitivity to modulation by zinc. Comparison of zinc- and diazepam-induced modulation of GABA-induced current responses in the same cells revealed an inverse correlation between these two modulators. No differences, however, were observed with the GABA(A) receptor modulators loreclezole, allopregnanolone, and pentobarbital. Thus, IMR-32 cells maintained in culture are heterogeneous in terms of expression of GABA(A) receptor isoforms. PMID- 10797555 TI - Partial sciatic nerve ligation induced more dramatic increase of neuropeptide Y immunoreactive axonal fibers in the gracile nucleus of middle-aged rats than in young adult rats. AB - Neuropeptide changes in primary sensory neurons caused by partial nerve injury are likely involved in the development of neuropathic pain. In this study, using immunocytochemistry, we examined neuropeptide Y (NPY) expression in lumbar dorsal root ganglion (DRG) cells of young adult (2-3 months old) and middle-aged (8-10 months old) rats 4 weeks after partial sciatic nerve ligation (PSNL). Significantly higher NPY immunoreactivity was induced in the injured side DRG neurons, the dorsal horn and the gracile nuclei in middle-aged rats than in young rats. Using combined fluorescent dye tracing and NPY immunostaining, we found in middle-aged rats that 46% injured DRG neurons projected to the gracile nucleus and 45% of injured neurons were also NPY-IR, whereas 42% spared DRG neurons projected to the gracile nucleus and 18% of spared neurons were also NPY-IR. Thus PSNL induces NPY up-regulation in spared as well as injured DRG neurons, both contribute to the increased NPY immunoreactivity in the gracile nucleus in the middle-aged rats. The more dramatic increase of NPY in DRG neurons of middle-aged rats after PSNL shows that the responses to partial nerve injury are age dependent, that suggests a possible relevance to the higher incidence of neuropathic pain in human middle age. PMID- 10797554 TI - Rescue of lesioned adult rat spinal motoneurons by adenoviral gene transfer of glial cell line-derived neurotrophic factor. AB - Glial cell line-derived neurotrophic factor (GDNF) has been shown to protect cranial and spinal motoneurons, that suggests potential uses of GDNF in the treatment of spinal cord injury and motor neuron diseases. We examined neuroprotective effect of human GDNF encoded by an adenovirus vector (AxCAhGDNF) on the death of lesioned adult rat spinal motoneurons. The seventh cervical segment (C7) ventral and dorsal roots and dorsal root ganglia of adult Fisher 344 rats were avulsed, and AxCAhGDNF, AxCALacZ (adenovirus encoding beta galactosidase gene) or PBS was inoculated in C7 vertebral foramen. One week after the avulsion and treatment with AxCALacZ, the animals showed expression of beta galactosidase activity in lesioned spinal motoneurons. Animals avulsed and treated with AxCAhGDNF showed intense immunolabeling for GDNF in lesioned spinal motoneurons and expression of virus-induced human GDNF mRNA transcripts in the lesioned spinal cord tissue. Nissl-stained cell counts revealed that the treatment with AxCAhGDNF significantly prevented the loss of lesioned ventral horn motoneurons 2 to 8 weeks after avulsion, as compared to AxCALacZ or PBS treatment. Furthermore, the AxCAhGDNF treatment ameliorated choline acetyltransferase immunoreactivity in the lesioned motoneurons after avulsion. These results indicate that the adenovirus-mediated gene transfer of GDNF may prevent the degeneration of motoneurons in adult humans with spinal cord injury and motor neuron diseases. PMID- 10797556 TI - Immunostaining of ganglioside GD1b, GD3 and GM1 in rat cerebellum: cellular layer and cell type specific associations. AB - We have studied the cellular distribution of gangliosides GD1b, GD3 and GM1 in rat cerebellum by immunostaining, using monoclonal antibodies and confocal microscopy. Antibodies against astroglial, neuronal and synaptic vesicle associated molecules were used for colocalization analyses. In the gray matter, the anti-GD1b antibody stained thin strands in the molecular layer (ML), interpreted as Bergman glia fibers based on colocalized staining with anti-glial fibrillary acidic protein (GFAP). The neuropil in the granule (GL) and Purkinje (PL) cell layers was also anti-GD1b positive. The anti-GD3 antibody stained the ML, the neuropil in the GL and PL and also the granule and Purkinje cell bodies, appearing intracytoplasmically and vesicle associated. Anti-GD1b and anti-GD3 staining in the GL glomeruli were colocalized with anti-synaptophysin staining. The anti-GM1 antibody stained cell bodies in the ML but they could not be characterized in colocalization experiments. The GL and PL were not stained with the anti-GM1 antibody. In the white matter, different staining patterns were seen for the gangliosides, the anti-GM1 staining being the most intense. This study shows cellular layer and cell type specific associations of the investigated gangliosides and localization of GD1b and GD3 at synaptic sites, warranting further studies on their role in synaptic mechanisms. PMID- 10797557 TI - Alterations of intracellular calcium homeostasis and mitochondrial function are involved in ruthenium red neurotoxicity in primary cortical cultures. AB - Ruthenium red (RR) is a polycationic dye that induces neuronal death in vivo and in primary cultures. To characterize this neurotoxic action and to determine the mechanisms involved, we have analyzed the ultrastructural alterations induced by RR in rat cortical neuronal cultures and measured its effect on cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) and on mitochondrial function. RR produced a dose-dependent, progressive disruption of neurites and plasma membrane of neuronal somata after 8-24 hr of incubation. RR caused also an elevation of both the basal [Ca(2+)](i) and its maximal levels after K(+) depolarization. Mitochondrial oxidative function, assessed by reduction of 3-(4,5-dimethylthiazol 2-yl)-2, 5-diphenyltetrazolium bromide and by changes in dihydrorhodamine-123 fluorescence, was significantly diminished after treatment with RR, both in cultured neurons and in isolated brain mitochondria. La(3+) did not prevent but rather potentiated RR-induced cell death. Glutamate receptor antagonists also failed to prevent RR neurotoxicity. Apoptotic electron microscope images were not observed, and protein synthesis inhibitors did not show any protective effect. It is concluded that RR penetrates neurons and that its neurotoxic damage probably is due to intracellular Ca(2+) dishomeostasis and disruption of mitochondrial oxidative function. These results enhance our understanding of the intracellular mechanisms underlying neuronal death. PMID- 10797558 TI - Metabolic stress in PC12 cells induces the formation of the endogenous dopaminergic neurotoxin, 3,4-dihydroxyphenylacetaldehyde. AB - 3,4-Dihydroxyphenylacetaldehyde (DOPAL) has been reported to be a toxic metabolite formed by the oxidative-deamination of dopamine (DA) catalyzed by monoamine oxidase. This aldehyde is either oxidized to 3,4-dihydroxyphenylacetic acid (DOPAC) by aldehyde dehydrogenase, an NAD-dependent enzyme or reduced to 3, 4-dihydroxyphenylethanol (DOPET) by aldehyde or aldose reductase. In the present study we examined whether levels of DOPAL are elevated by inhibition of the mitochondrial respiratory chain. Using inhibitors of mitochondrial complexes I, II, III and IV we found that inhibition of complex I and III increased levels of DOPAL and DOPET. Nerve growth factor-induced differentiation of PC12 cells markedly potentiated DOPAL and DOPET accumulation in response to metabolic stress. DOPAL was toxic to differentiated PC12 as well as to SK-N-SH cell lines. Because complex I dysfunction has been implicated in the pathogenesis of Parkinson's disease, the accumulation of DOPAL may explain the vulnerability of the dopaminergic system to complex I inhibition. The rapid appearance of DOPAL and DOPET after inhibition of complex I may be a useful early index of oxidative stress in DA-forming neurons. PMID- 10797559 TI - Tau hyperphosphorylation in apolipoprotein E-deficient and control mice after closed head injury. AB - Apolipoprotein E (apoE)-deficient mice have learning and memory impairments that are associated with specific neurochemical changes and hyperphosphorylation of distinct epitopes of the cytoskeletal protein tau. Furthermore, such mice are highly susceptible to the sequelae of brain trauma and their ability to recover from head injury is impaired. In the present study we investigated the extent that the neuronal maintenance and repair impairments of apoE-deficient mice are related to aberrations at the tau phosphorylation level. This was pursued by subjecting control and apoE-deficient mice to closed head injury (CHI) and examination, utilizing immunoblot assays, of the resulting effects on tau phosphorylation. The results thus obtained revealed that tau of apoE-deficient mice is hyperphosphorylated before CHI and that this insult results in transient tau hyperphosphorylation, whose extent and time course in the two mouse groups varied markedly. Tau hyperphosphorylation in the injured controls was maximal by about 4 hr after injury and reverted to basal levels by 24 hr. In contrast, almost no head injury-induced tau hyperphosphorylation was observed in the apoE deficient mice at 4 hr after injury. Some tau hyper-phosphorylation was detected in the head-injured apoE-deficient mice after longer time intervals, but its extent was markedly lower than the maximal values obtained in the head injured controls. These findings show that the chronic neuronal impairments brought about by apoE deficiency and the acute response to head injury are both associated with hyperphosphorylation of the same tau domain and that the ability of apoE deficient mice to mount the acute tau hyperphosphorylation response to head injury is impaired. PMID- 10797560 TI - APP carboxyl-terminal fragment without or with abeta domain equally induces cytotoxicity in differentiated PC12 cells and cortical neurons. AB - Mutations in the beta-amyloid precursor protein (APP) gene cause familial Alzheimer's disease (AD). Although amyloid beta peptide (Abeta) is the principal constituent of senile plaques in AD, other cleavage products of APP are also implicated in playing a role in the pathogenesis of AD. C-terminal fragments of APP (APP-CTs), that contain complete Abeta sequence, are found in neuritic plaques, neurofibrillary tangles and the cytosol of lymphoblastoid cells obtained from AD patients. Our previous report demonstrated that APP-CT105 causes death of differentiated PC12 cells and cultured rat cortical neurons (Kim and Suh [1996] J. Neurochem. 67:1172-1182) and induces strong inward currents in Xenopus oocyte (Fraser et al., [1996] J. Neurochem. 66:2034-2040). In the present study, to investigate which domain of APP-CT105 is responsible for the neurotoxicity, we have made deletion mutants of APP-CT105 without Abeta and transmembrane domain (TM) or without NPTY domain, a putative endocytosis signaling sequence, using the PCR-amplified strategy and the recombinant GST-fusion protein strategy. The effect on cell survival of the deletion mutants of APP-CT105 (8 microM) was then determined by the LDH and MTT assay. We found that C-terminal fragment without NPTY significantly causes cell death in NGF-differentiated PC12 cells and cultured rat cortical neurons. This finding suggests that NPTY may not play an important role in APP-CT105 mediated neurotoxicity. We found, however, that C terminal fragment without Abeta and TM significantly induces neuronal cell death. Our results suggest that in addition to Abeta, C-terminal fragment of APP without Abeta and TM domain itself may also participate in the neuronal degeneration in AD. PMID- 10797561 TI - Biochemical characterization of a nuclear factor that binds to NF1-like elements in the rat p53 promoter. AB - We previously reported that two nuclear factor 1-like elements mediated the transcription of the rat p53 gene. A 40-kDa protein was shown to bind to these elements, which was different from common NF1 family proteins. In this study, the biochemical properties of the 40-kDa binding protein were investigated. The metal ion dependency of the protein was examined with various chelators; the protein was proved to require Mg(2+) for maximum DNA-binding activity. The binding protein was highly resistant to ionic strength and denaturant. The protein-DNA complex was reduced at high NaCl concentration, but residual DNA-binding activity remained. Even 2 M urea did not completely eliminate the formation of protein-DNA complex. DNA-binding activity of the protein was also stable at high temperature. Treatment of the protein-DNA complex with increasing concentrations of proteinase K or trypsin demonstrated the existence of a protease-resistant DNA-bound core. These biochemical properties provide new insight into the 40-kDa NF1-like nuclear factor. PMID- 10797562 TI - Transforming JB6 cells exhibit enhanced integrin-mediated adhesion to osteopontin. AB - Transformation of preneoplastic epidermal JB6 cells with tumor promoter 12-O tetradecanoyl-phorbol-13-acetate (TPA) is an in vitro model of late-stage tumor promotion. Osteopontin (OPN) is a secreted, adhesive protein that is highly expressed in JB6 cells with TPA treatment, and its expression persists for at least 4 days, which is the time required for subsequent expression of transformed phenotype. These observations suggest that OPN may play a role in promoting JB6 cell transformation. To function in transformation of JB6 cells, OPN must bind to the surface of the JB6 cell and subsequently signal within the cell. Therefore, we investigated whether JB6 cells adhere to OPN and, if so, to which surface receptors. TPA-treated JB6 cells had significantly (P < 0.05) increased adherence to OPN compared with dimethylsulfoxide-treated control cells. Enhanced attachment of JB6 cells to OPN was also observed after treatment with another tumor promoter phorbol dibutyrate but not with nontumor promoters (phorbol and 1alpha,25 dihydroxyvitamin D(3)), suggesting that tumor promoters specifically modulate attachment to OPN. The argininylglycylaspartic acid (RGD) cell-binding region of OPN mediates attachment of TPA-treated JB6 cells because RGD, but not argininylglycylglutamic acid (RGE), peptides inhibited adherence of these cells to OPN in a dose-dependent manner. Flow cytometric analyses, blocking adhesion assay using anti-alpha(v) antibody, and co-immunoprecipitation assay all indicated that TPA-treated cells had similar levels of alpha(v) and beta(5) but decreased levels of beta(1) compared with untreated cells and that cell adhesion to OPN is most likely mediated through the alpha(v)beta(5). Furthermore, calphostin C, a specific protein kinase C (PKC) inhibitor, decreased TPA-treated JB6 cell adhesion to OPN by 50%, suggesting that TPA increased integrin affinity or avidity for OPN through a PKC-mediated pathway. Collectively, these results indicate that transforming JB6 cells adhere to OPN through its RGD sequence. The most likely OPN receptor is the alpha(v)beta(5) integrin, which increases the affinity or avidity for OPN through a PKC-dependent pathway rather than increasing the number of receptors. PMID- 10797563 TI - Truncated activin type II receptor inhibits erythroid differentiation in K562 cells. AB - Two receptor serine/threonine kinases (types I and II) have been identified as signaling transducing activin receptors. We studied the possibility of inhibiting activin A-dependent differentiation in K562 cells, using a dominant negative mutant of type II receptor. A vector was constructed expressing activin type II truncated receptor (ActRIIa) that lacks the cytoplasmic kinase domain. Since activin type I and II receptors form heteromeric complexes for signaling, the mutant receptors compete for binding to endogenous receptors, hence acting in a dominant negative fashion. K562 cells were stably transfected with ActRIIa, and independent clones were expanded. The truncated cDNA was integrated into the genome of the transfectants, as shown by polymerase chain reaction; and the surface expression of truncated receptors was shown by affinity cross-linking with (125)I-activin A. In wild-type K562 cells, activin A induced erythroid differentiation and cells started to express hemoglobins. In transfected cells expressing ActRIIa, the induction of erythroid differentiation was abrogated and less than 10% of cells were hemoglobin-containing cells after culture with activin A. Further transfection with wild-type type II receptors rescued the mutant phenotype of these transfectants, indicating that the effect of ActRIIa is dominant negative. In addition, phosphorylation of the cytoplasmic kinase domain of the type II receptor in vitro confirms the autophosphorylation of this portion of the receptor. Therefore, induction of erythroid differentiation in vitro is mediated through the cell surface activin receptor, and interference with this receptor signaling inhibits this process of differentiation in K562 cells. PMID- 10797564 TI - Annexin II is the membrane receptor that mediates the rapid actions of 1alpha,25 dihydroxyvitamin D(3). AB - 1alpha,25-Dihydroxyvitamin D(3) has been shown to exert its effects by both genomic (minutes to hours) and rapid (seconds to minutes) mechanisms. The genomic effects are mediated by interaction with the nuclear vitamin D receptor. We show that the vitamin D analog, [(14)C]-1alpha,25-dihydroxyvitamin D(3) bromoacetate, is specifically bound to a protein (molecular weight 36 kDa) in the plasma membrane of rat osteoblastlike cells (ROS 24/1). The plasma membrane protein labeled with the bromoacetate analog was identified as annexin II by sequence determination and Western blot. Partially purified plasma membrane proteins (PI 6.9-7.4) and purified annexin II exhibited specific and saturable binding for [(3)H]-1alpha, 25-dihydroxyvitamin D(3). Antibodies to annexin II inhibited [(14)C]-1alpha,25-dihydroxyvitamin D(3) bromoacetate binding to ROS 24/1 plasma membranes, immunoprecipitated the ligand-protein complex, and inhibited 1alpha,25 dihydroxyvitamin D(3)-induced increases in intracellular calcium in ROS 24/1 cells. The results indicate that annexin II may serve as a receptor for rapid actions of 1alpha, 25-dihydroxyvitamin D(3). PMID- 10797565 TI - Effects of increased ambient pressure on colon cancer cell adhesion. AB - Forces such as strain modulate intestinal epithelial biology. Shear and pressure influence other cells. The effects of pressure on human colon cancer cells are poorly understood. Increasing ambient pressure for 30 min by 15 mm Hg over atmospheric stimulated adhesion to matrix proteins of four human colon cancer cell lines and primary cells from three human colon cancers, but not bovine aortic smooth-muscle cells. This effect was energy dependent and cation dependent (blocked by azide and chelation), accompanied by tyrosine phosphorylation of intracellular proteins including focal adhesion kinase, and blocked by tyrosine kinase inhibition (genistein, tyrphostin, and erbstatin) and a functional antibody to the beta1 integrin subunit. Although pressure stimulated adhesion even in a balanced salt solution, baseline and pressure-stimulated adhesion were each substantially diminished in the absence of serum. These data suggest that relatively low levels of increased pressure may stimulate malignant colonocyte adhesion by a cation-dependent beta1-integrin-mediated mechanism, perhaps via focal adhesion kinase-related tyrosine phosphorylation. In addition to elucidating another aspect of physical force regulation of colonocyte biology, these findings may be relevant to the effects of increased pressure engendered by colonic peristalsis, surgical manipulation, or laparoscopic surgery on colon cancer cell adhesion. PMID- 10797566 TI - Subcellular localization of aldolase B. AB - The localization of the aldolase B isozyme was determined immunohistochemically in rat kidney and liver using a polyclonal antibody. Aldolase B was preferentially localized in a nuclear region of hepatocytes from the periportal region and was absent in those from the perivenous region. Aldolase B was also preferentially localized in the proximal tubules and was absent in other structures of the renal cortex as well as in the renal medulla. Using reflection confocal microscopy, the enzyme was preferentially localized in a nuclear position in liver and renal cells, which was similar to the cellular and intracellular location found for the gluconeogenic enzyme fructose-1,6 bisphosphatase (Saez et al. [1996] J. Cell. Biochem. 63:453-462). Subcellular fractionation studies followed by enzyme activity assays revealed that aldolase activity was associated with subcellular particulate structures. Overall, the data suggest that different aldolase isoenzymes are needed in the glycolytic and gluconeogenic pathways. PMID- 10797567 TI - Inhibition of UV irradiation-induced oxidative stress and apoptotic biochemical changes in human epidermal carcinoma A431 cells by genistein. AB - Ultraviolet (UV) light is a strong apoptotic trigger that can induce a caspase dependent biochemical change in cells. We previously showed that UV irradiation can elicit caspase-3 activation and the subsequent cleavage and activation of p21 activated kinase 2 (PAK2) in human epidermal carcinoma A431 cells. We report that genistein, an isoflavone compound with known inhibitory activities to protein tyrosine kinases (PTKs) and topoisomerase-II (topo-II), can prevent UV irradiation-induced apoptotic biochemical changes (DNA fragmentation, caspase-3 activation, and cleavage/activation of PAK2) in A431 cells. Surprisingly, two typical PTK inhibitors (tyrphostin A47 and herbimycin A) and three known topo-II inhibitors (etoposide, daunorubicin, and novomycin) had no effect on UV irradiation-induced apoptotic biochemical changes, suggesting that the inhibitory effect of genistein is not dependent on its property as a PTK/topo-II inhibitor. In contrast, azide, a reactive oxygen species (ROS) scavenger, could effectively block the UV irradiation-induced apoptotic cell responses. Flow cytometric analysis using the cell-permeable dye 2',7'-dichlorofluorescin diacetate as an indicator of the generation of ROS showed that UV irradiation caused increase of the intracellular oxidative stress and that this increase could be abolished by azide, suggesting that oxidative stress plays an important role in mediating the apoptotic effect of UV irradiation. Importantly, the UV irradiation-induced oxidative stress in cells could be significantly attenuated by genistein, suggesting that impairment of ROS formation during UV irradiation is responsible for the antiapoptotic effect of genistein. Collectively, our results demonstrate the involvement of oxidative stress in the UV irradiation-induced caspase activation and the subsequent apoptotic biochemical changes and show that genistein is a potent inhibitor for this process. PMID- 10797568 TI - Multiple domains of occludin are involved in the regulation of paracellular permeability. AB - Tight junctions form selective paracellular diffusion barriers that regulate the diffusion of solutes across epithelia and constitute intramembrane diffusion barriers that prevent the intermixing of apical and basolateral lipids in the extracytoplasmic leaflet of the plasma membrane. In MDCK cells, previous expression experiments demonstrated that occludin, a tight junction protein with four transmembrane domains, is critically involved in both of these tight junction functions and that its COOH-terminal cytoplasmic domain is of functional importance. By expressing mutant and chimeric occludin that exert a dominant negative effect on selective paracellular diffusion, we now demonstrate that the extracytoplasmic domains and at least one of the transmembrane domains are also critically involved in selective paracellular permeability. Multiple domains of occludin are thus important for the regulation of paracellular permeability. Expression of chimeras containing at least one transmembrane domain of occludin also resulted in an enhanced intracellular accumulation of claudin-4, another transmembrane protein of tight junctions, suggesting that the two proteins may cooperate in the regulation of paracellular permeability. PMID- 10797569 TI - beta1B integrin subunit contains a double lysine motif that can cause accumulation within the endoplasmic reticulum. AB - Human epidermal keratinocytes are one of the few cell types that express the beta1B splice variant of the beta1 integrin subunit. Although in transfection experiments beta1B acts as a dominant negative inhibitor of cell adhesion, we found that beta1B was expressed at very low levels in keratinocytes, both in vivo and in culture, and had a predominantly cytoplasmic distribution, concentrated within the endoplasmic reticulum. To examine why beta1B accumulated in the cytoplasm, we prepared chimeras between CD8alpha and the beta1A and beta1B integrin cytoplasmic domains. In transfected HeLa cells, both constructs reached the cell surface but the rate of maturation of the beta1B chimera was considerably retarded relative to beta1A. The beta1B cytoplasmic domain contains two lysine residues that resemble the double lysine motif characteristic of many proteins that are resident within the endoplasmic reticulum. Mutation of each lysine individually to serine had no effect on CD8beta1B maturation, but when both residues were mutated the rate of CD8beta1B maturation increased to that of CD8beta1A. Further analysis of beta1B function in keratinocytes must, therefore, take into account the low abundance of the isoform relative to beta1A and the potential for beta1B to accumulate in the endoplasmic reticulum. PMID- 10797570 TI - Natural metabolites of 1alpha,25-dihydroxyvitamin D(3) retain biologic activity mediated through the vitamin D receptor. AB - 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)), the active metabolite of vitamin D, mediates many of its effects through the intranuclear vitamin D receptor (VDR, NR1I1), that belongs to the large superfamily of nuclear receptors. Vitamin D receptor can directly regulate gene expression by binding to vitamin D response elements (VDREs) located in promoter or enhancer regions of various genes. Although numerous synthetic analogs of 1alpha,25(OH)(2)D(3) have been analysed for VDR binding and transactivation of VDRE-driven gene expression, the biologic activity of many naturally occurring metabolites has not yet been analyzed in detail. We therefore studied the transactivation properties of 1alpha,24R, 25-trihydroxyvitamin D(3) (1alpha,24R,25(OH)(3)D(3)), 1alpha, 25 dihydroxy-3-epi-vitamin D(3) (1alpha,25(OH)(2)-3-epi-D(3)), 1alpha,23S,25 trihydroxyvitamin D(3) (1alpha,23S,25(OH)(3)D(3)), and 1alpha-hydroxy-23-carboxy 24,25,26,27-tetranorvitamin D(3) (1alpha(OH)-24,25,26,27-tetranor-23-COOH-D(3); calcitroic acid) using the human G-361 melanoma cell line. Cells were cotransfected with a VDR expression plasmid and luciferase reporter gene constructs driven by two copies of the VDRE of either the mouse osteopontin promoter or the 1alpha,25(OH)(2)D(3) 24-hydroxylase (CYP24) promoter. Treatment with 1alpha,25(OH)(2)D(3) or the metabolites 1alpha,24R,25(OH)(3)D(3), 1alpha,25(OH)(2)-3-epi-D(3), and 1alpha,23S,25(OH)(3)D(3) resulted in transactivation of both constructs in a time- and dose-dependent manner, and a postitive regulatory effect was observed even for calcitroic acid in the presence of overexpressed VDR. The metabolites that were active in the reporter gene assay also induced expression of CYP24 mRNA in the human keratinocyte cell line HaCaT, although with less potency than the parent hormone. A ligand-binding assay based on nuclear extracts from COS-1 cells overexpressing human VDR demonstrated that the metabolites, although active in the reporter gene assay, were much less effective in displacing [(3)H]-labeled 1alpha,25(OH)(2)D(3) from VDR than the parent hormone. Thus, we report that several natural metabolites of 1alpha,25(OH)(2)D(3) retain significant biologic activity mediated through VDR despite their apparent low affinity for VDR. PMID- 10797571 TI - Stimulatory effect of regucalcin on ATP-dependent Ca(2+) uptake activity in rat liver mitochondria. AB - The effect of Ca(2+)-binding protein regucalcin on Ca(2+)-ATPase activity in isolated rat liver mitochondria was investigated. The presence of regucalcin (0.1, 0.25, and 0.5 microM) in the enzyme reaction mixture led to a significant increase in Ca(2+)-ATPase activity. Regucalcin significantly stimulated ATP dependent (45)Ca(2+) uptake by the mitochondria. Ruthenium red (10(-5) M) or lanthanum chloride (10(-4) M), an inhibitor of mitochondrial Ca(2+) uptake, completely inhibited regucalcin (0.25 microM)-increased mitochondrial Ca(2+) ATPase activity and (45)Ca(2+) uptake. The effect of regucalcin (0.25 microM) in increasing Ca(2+)-ATPase activity was completely inhibited by the presence of digitonin (10(-2)%), a solubilizing reagent of membranous lipids, or vanadate (10(-5) M), an inhibitor of phosphorylation of ATPase. The activatory effect of regucalcin (0.25 microM) on Ca(2+)-ATPase activity was not further enhanced in the presence of dithiothreitol (2.5 mM), a protecting reagent of the sulfhydryl (SH) group of the enzyme, or calmodulin (0.60 microM), a modulator protein of Ca(2+) action that could increase mitochondrial Ca(2+)-ATPase activity. The present study demonstrates that regucalcin can stimulate Ca(2+) pump activity in rat liver mitochondria, and that the protein may act on an active site (SH group) related to phosphorylation of mitochondrial Ca(2+)-ATPase. PMID- 10797572 TI - Regulation of (1-3)-beta-glucan-stimulated Ca(2+) influx by protein kinase C in NR8383 alveolar macrophages. AB - Stimulation of (1-3)-beta-glucan receptors results in Ca(2+) influx through receptor-operated channels in alveolar macrophages (AMs), but the mechanism(s) regulating Ca(2+) influx is still undefined. In this study we investigated the role of protein kinase C (PKC) regulation of Ca(2+) influx in the NR8383 AM cell line using the particulate (1-3)-beta-glucan receptor agonist zymosan. PKC inhibition with calphostin C (CC) or bisindolymaleimide I (BSM) significantly reduced zymosan-induced Ca(2+) influx, whereas activation of PKC with phorbol-12 myristate 13-acetate (PMA) or 1, 2-dioctanoyl-sn-glycerol (DOG) mimicked zymosan, inducing a concentration-dependent Ca(2+) influx. This influx was dependent on extracellular Ca(2+) and inhibited by the receptor-operated Ca(2+) channel blocker SK&F96365, indicating that zymosan and PKC activate Ca(2+) influx through a similar pathway. NR8383 AMs expressed one new PKC isoform (delta) and two atypical PKC isoforms (iota and lambda), but conventional PKC isoforms were not present. Stimulation with zymosan resulted in a translocation of PKC-delta from the cytosol to the membrane fraction. Furthermore, inhibition of protein tyrosine kinases (PTKs) with genistein prevented zymosan-stimulated Ca(2+) influx and PKC delta translocation. These results suggest that PKC-delta plays a critical role in regulating (1-3)-beta-glucan receptor activated Ca(2+) influx in NR8383 AMs and PKC-delta translocation is possibly dependent on PTK activity. PMID- 10797573 TI - Vimentin expression in human squamous carcinoma cells: relationship with phenotypic changes and cadherin-based cell adhesion. AB - Phenotypic changes resembling an epithelial-to-mesenchymal transition often occur as epithelial cells become tumorigenic. Two proteins that have been implicated in this process are vimentin and N-cadherin. In this study, we sought to establish a link between expression of vimentin and N-cadherin as oral squamous epithelial cells undergo a morphologic change resembling an epithelial-to-mesenchymal transition. We found that N-cadherin and vimentin did not influence the expression of one another. PMID- 10797574 TI - Cdk5/p25(nck5a) interaction with synaptic proteins in bovine brain. AB - Cyclin-dependent kinase 5 (Cdk5) exists in large multimeric complexes, but its function and binding partners in these complexes are unclear. We explored these issues by chromatographic and immunochemical analyses of Cdk5 and p25(nck5a) (a neuronal Cdk5 activator) and their associated proteins from bovine brain. Mono-S column enzyme eluates were divided into three fractions and analyzed by gel filtration. The majority of p25(nck5a) from Mono-S fractions I, II, and III eluted from the gel filtration column at approximately 60, 200, and 400 kDa, respectively, and Cdk5 was abundant in fractions >400 kDa. We characterized these macromolecular structures by immunoprecipitating p25(nck5a), followed by a second immunoprecipitation of remaining unbound proteins using a Cdk5 antibody. The p25(nck5a) immunoprecipitates showed association with Cdk5. Amphiphysin was detected in the 400-kDa complex and synapsin I in the >400 kDa structure. The Cdk5 immunoprecipitates, however, revealed abundant retained Cdk5 but no remaining p25(nck5a), indicating that Cdk5 in macromolecular structures is mostly unassociated with p25(nck5a). Thus, we demonstrate: an amphiphysin-associated 400 kDa Cdk5/p25(nck5a) complex, a synapsin I-associated >400-kDa Cdk5/p25(nck5a) complex, and nck5a-free Cdk5 complexes (200 to >400 kDa). Amphiphysin acts as a Cdk5/p25(nck5a) substrate in the 400-kDa complex and we speculate that Cdk5/p25(nck5a) participates in amphiphysin-mediated endocytosis. PMID- 10797575 TI - Discrete region of the insulin receptor carboxyl terminus plays key role in insulin action. AB - In the present study, we attempted to determine the importance of a 23-amino-acid sequence within the carboxyl terminus of the human insulin receptor (IR) molecule in modulating insulin action in Chinese hamster ovary cells. Stable expression of a minigene encoding the receptor fragment led to an increase in insulin-induced IR autophosphorylation that was 2.4-fold higher when compared to that of IR expressing cells transfected with empty vector. Insulin-stimulated downstream signaling was also significantly elevated in cells expressing the minigene. It was found that expression of the minigene had no effect toward insulin-like growth factor I receptor kinase activity and function. These results indicate that the IR carboxyl terminus contains a motif that acts as a physiologic modulator of insulin signaling. J. Cell. Biochem. 78:160-169, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10797577 TI - Freedom of the press and the public's right to know. PMID- 10797576 TI - Genomic organization, promoter analysis, and chromosomal mapping of the mouse gene encoding Cdk9. AB - Cdk9, previously known as PITALRE, belongs to the Cdc2 family of protein kinases. We report the isolation and characterization of the complete gene coding for the murine Cdk9 protein. The gene contains seven exons spanning over 6 kb of genomic DNA, and the exon/intron boundaries conformed to the GT/AG rule. The Cdk9 gene mapped on mouse chromosome 2, which is consistent with the known region of synteny with human chromosome 9q34.1. The length of the individual exons ranged from 82 to 850 bp, and introns ranged from 452 to 1,465 bp. The further 5' flanking region of the gene showed features of a housekeeping promoter, such as the lack of a canonical TATA box and the presence of a CCAAT box as well as several GC boxes, which are potential binding sites for numerous transcription factors. Additionally, we performed a basic analysis of the transcriptional activity of the promoter and found that the 364 bp of Cdk9 5' flanking region were able to elicit high transcriptional levels of a luciferase reporter gene in NIH3T3 cells. This study provides the molecular basis for understanding the transcriptional control of the Cdk9 gene, and could serve to facilitate the molecular genetic investigation of Cdk9 function during mouse embryonal development. PMID- 10797578 TI - Bladder outlet obstruction in women: definition and characteristics. AB - The prevalence of bladder outlet obstruction in women is unknown and most probably has been underestimated. Moreover, there are no standard definitions for the diagnosis of bladder outlet obstruction in women. Our study was conducted to define as well as to examine the clinical and urodynamic characteristics of bladder outlet obstruction among women referred for evaluation of voiding symptoms. Bladder outlet obstruction was defined as a persistent, low, maximum "free" flow rate of <12 mL/s in repeated non-invasive uroflow studies, combined with high detrusor pressure at a maximum flow (p(det.Q)(max) >20 cm H(2)O) during detrusor pressure-uroflow studies. A urodynamic database of 587 consecutive women identified 38 (6.5%) women with bladder outlet obstruction. The mean age of the patients was 63.9 +/- 17.5 years. The mean maximum "free" flow, voided volume, and residual urinary volume were 9.4 +/-3.9 mL/s, 144. 9 +/- 72.7 mL, and 86.1 +/ 98.8 mL, respectively. The mean p(det. Q)(max) was 37.2 +/- 19.2 cm H(2)O. Previous anti-incontinence surgery and severe genital prolapse were the most common etiologies, accounting for half of the cases. Other, less common, etiologies included urethral stricture (13%), primary bladder neck obstruction (8%), learned voiding dysfunction (5%), and detrusor external sphincter dyssynergia (5%). Symptomatology was defined as mixed obstructive and irritative in 63% of the patients, isolated irritative in 29%, and isolated obstructive in other 8%. In conclusion, bladder outlet obstruction in women appears to be more common than was previously recognized, occurring in 6.5% of our patients. Micturition symptoms relevant to bladder outlet obstruction are non-specific, and a full urodynamic evaluation is essential in making the correct diagnosis and formulating a treatment plan. PMID- 10797579 TI - Intra- and inter-investigator variation in the analysis of pressure-flow studies in men with lower urinary tract symptoms. AB - The objective of this study was to assess the intra- and inter-investigator variation in the analysis of pressure-flow studies that were performed in men with lower urinary tract symptoms. Two hospitals were involved in this study. In each hospital 100 PFS were selected. Photocopies of printouts of all PFS were analyzed manually by six experienced investigators, including determination of P(detQmax) and Q(max). Afterward, all 200 PFS were analyzed again in a different order. For each P(detQmax) and accompanying Q(max) the AG-number was calculated. With these AG numbers, the intra-investigator SD, the inter-investigator SD and the intra- and inter-investigator SD combined were calculated. The intra- and inter-investigator SD combined was 10.7. This implies that if one investigator analyzes a PFS once and determines an AG number of 40, another investigator may determine an AG number between 40 +/- 2. 77*10.7 = 10-70, using a 95% confidence interval. The inter-investigator SD was 10.0 and the intra-investigator SD was 3.7. The reproducibility of the manual analysis of urodynamic studies is moderate owing to a considerable intra- and inter- investigator variation. This is mostly caused by the substantial intra-investigator variation. PMID- 10797580 TI - Can persisting detrusor hyperreflexia be predicted after transurethral prostatectomy for benign prostatic hypertrophy? AB - Detrusor hyperreflexia (DH) is frequently found in patients with benign prostatic hypertrophy (BPH) and persists in 30-50% of patients after successful removal of bladder neck obstruction by transurethral prostatectomy (TUR-P) or surgical enucleation of the prostate. It would be beneficial for surgeons to be able to identify patients who are at risk of persistent post-operative urinary irritation symptoms and DH. Twenty-three patients who showed DH pre-operatively were included in this study. Of these 23 patients, four had neurogenic bladder because of previous cerebrovascular disease. The other 19 patients were considered to have DH because of BPH. These 19 patients were classified according to their cystometry chart patterns. Pattern 1 was the continual sporadic onset and offset of DH, pattern 2 was a single episode of DH at a bladder volume of <160 mL, and pattern 3 was a single DH episode at a bladder volume >160 mL. Preoperative single-photon emission computed tomography (SPECT) was performed on 14 patients. Cystometric findings at 3 to 6 months after surgery were compared with the pre operative findings. Four of the six patients with pattern 2 (67%) and all patients with pattern 3 (100%) showed an absence of DH after surgery. In contrast, all five patients with pattern 1 and all four patients with neurogenic bladder showed persistent DH. Compared with pattern 3 patients, pattern 1 patients more frequently complained of urgency before surgery, and their symptoms and uroflowmetry parameters did not improve afterward. Among 14 patients who had pre-operative SPECT, all eight patients with low cerebral blood flow in the frontal region showed persisting DH. Conversely, all six patients with normal SPECT results showed no DH after surgery. When DH occurs repeatedly (pattern 1) or occurs at a bladder volume of <160 mL (pattern 2), there is a greater risk of post-operative irritation symptoms. Abnormal SPECT findings can also predict the post-operative persistence of DH. Combing these two pre-operative examinations allows us to predict better post-operative DH in patients with BPH. PMID- 10797581 TI - Urethral obstruction in patients with nighttime wetting: urodynamic evaluation and outcome of surgical incision. AB - The aim of this study was to investigate urodynamic findings and the outcome of surgical incision in patients with nighttime wetting due to urethral obstruction. A total of 239 patients with nighttime wetting (157 males and 82 females; mean age, 8.7; range, 4-18 years) was studied. One hundred and ten patients had monosymptomatic enuresis alone and 129 both nighttime wetting and daytime symptoms, that is, the majority was not simple bedwetters, but had signs of bladder overactivity. All patients were first treated with bladder training and fluid intake after dinner was restricted. The non-responders were then treated with tricyclic antidepressants, anticholinergics, or intra-nasal desmopressin and conditioning therapy. Ninety-seven patients who did not respond to these conventional treatments were investigated by voiding cystourethrography, bougie a boule, and urodynamic study. Urethral obstruction was noted in 34 patients (14.2%); posterior urethral valves in three males, ring stricture of the bulbar urethra in 26 males and distal urethral stenosis in five females. Detrusor instability was noted in 30 (94%) of 32 patients. Three patients had <15 mL/s of maximum flow rate and none had post-void residual urine. Pressure at maximum flow (PQ(max)) was 106.5 +/- 26.3 cm H(2)O in pressure/flow study. After surgery, nighttime wetting was cured in 25 patients (73.5%), improved in four (11.8%). Detrusor instability disappeared in six of 16 patients studied (37.5%) and improved in 8 (50%), and PQ(max) decreased significantly after operation (P = 0. 0034). In conclusion, most of the patients with urethral obstruction have detrusor instability and high pressure voiding, and these conditions improved after operation. PMID- 10797582 TI - Anal sphincter electromyography after vaginal delivery: neuropathic insufficiency or normal wear and tear? AB - The study was performed to evaluate the potential role of vaginal delivery on innervation of the external anal sphincter (EAS) muscle. Forty-four women, 18 nulliparous and 26 of varying parity (1-4), without genitourinary prolapse, major urogynecological, anorectal, or neurological dysfunction were included. Participants' histories were evaluated by a questionnaire. Quantitative concentric needle electromyography (EMG) using multi-MUP analysis for sampling motor unit potentials (MUPs) in all volunteers, and "turn/amplitude" analysis for interference pattern (IP) analysis in 13 nulliparous and 23 parous women were applied. Pools of MUPs and IPs of parous and nulliparous women were formed and compared using the Mann-Whitney U test. Multiple linear regression analysis was used for evaluation of parity and obstetric variables. No difference between the groups was found in any MUP parameter, while a significant difference was found in two of five IP parameters. On multiple linear regression analysis, the number of deliveries was related to several MUP and IP parameters: the time elapsed since last delivery to MUP, and slight stress urinary incontinence to IP parameters. A group of parous women with (slight) stress incontinence had less "pathologic" MUP parameters, compared to those without. Vaginal delivery is indeed related to EAS muscle EMG abnormalities. However, these are minor and seem not to indicate loss of sphincter function. Our study casts some doubt on the commonly accepted preconception that significant damage to peripheral innervation of the EAS occurs even during uncomplicated deliveries. PMID- 10797583 TI - Urinary incontinence in both sexes: prevalence rates and impact on quality of life and sexual life. AB - The aim of this study was to determine the prevalence of urinary incontinence in both sexes in Austria and to assess its impact on quality of life and sexual function. Voluntary health examinations free of charge are regularly organized in the area of Vienna. From May 1998 to April 1999 we have included in this health examination an incontinence questionnaire containing 37 items, which was largely based on the Bristol female lower urinary tract symptoms (LUTS) questionnaire. This questionnaire asks in detail for various aspects of urinary incontinence/voiding problems, including the impact of urinary incontinence on quality of life and sexual function. In this questionnaire, urinary incontinence was defined as any involuntary loss of urine within the past 4 weeks. The data of 2,498 participants (1,262 women [f]: mean age: 49.7+/-13.6 years and 1,236 men [m]: 48.6+/-13.0 years; age range: 20-96 years) were analyzed. Overall, 26.3% of women and 5.0% of men reported on episodes of urinary incontinence during the past 4 weeks. Prevalence rates increased constantly with age in both sexes: 20-29 years: 4.1% (f), 1.7% (m); 30-39 years: 10.8% (f), 2.7% (m); 40-49 years: 22.9% (f); 3.9% (m); 50-59 years: 34.9% (f), 3.7% (m); 60-69 years: 36.9% (f), 7.6% (m); 70 years or older: 36.0%% (f), 11.5% (m). Overall, 65.7% of women and 58.3% of men stated that quality of life was affected by their incontinence status. A moderate or severe impairment was reported by 18.3% of women and 16.6% of men. Impairment of quality of life was related as statistically significant (P < 0.05) to frequency and degree of incontinence (irrespective of the type of incontinence), the impact on sexual function and need for pads or other incontinence devices. Patient gender, age, and the duration of incontinence had no effect (P > 0.05) on quality of life. An impairment of sexual life by urinary incontinence was stated by 25. 1% of women and 30.5% of women, respectively. Although only 65.7% of women and 58.3% men with urinary incontinence reported on an impairment of quality of life, these data underline the high prevalence and socioeconomic implications of this disorder. The impact of urinary incontinence on quality of life is significantly higher than on sexual function. PMID- 10797584 TI - Micturitional disturbance in a patient with neurosarcoidosis. AB - We report a case of neurosarcoidosis in which urodynamic studies showed neurogenic bladder dysfunction. A 30-year-old man began to have slowly progressive gait ataxia of vestibular origin, deafness, and hallucination, which developed into versive seizure and stupor. Brain computed tomography and magnetic resonance imaging showed the anteromedial frontal lobe lesion with mild ventricular enlargement. The cerebrospinal fluid examination revealed pleocytosis with raised total protein and angiotensin-converting enzyme levels. Endoscopic lung biopsy showed epithelioid granuloma. Oral prednislone (60 mg/day) ameliorated his symptoms. After tapering steroids, however, he developed urinary urgency, frequency, urge urinary incontinence, and a relapse of gait ataxia. The urodynamic study showed detrusor hyperreflexia. Prednisolone treatment again improved his urinary and neurological symptoms. The anteromedial frontal lobe lesion seems to be responsible for the micturitional disturbance in our patient with neurosarcoidosis. PMID- 10797586 TI - Comparison of different computer models of the neural control system of the lower urinary tract. AB - This paper presents a series of five models that were formulated for describing the neural control of the lower urinary tract in humans. A parsimonious formulation of the effect of the sympathetic system, the pre-optic area, and urethral afferents on the simulated behavior are included. In spite of the relative simplicity of the five models studied, behavior that resembles normal lower urinary tract behavior as seen during an urodynamic investigation could be simulated. The models were tested by studying their response to disturbances of the afferent signal from the bladder. It was found that the inhibiting reflex that results from including the sympathetic system or the pre-optic area (PrOA) only counteracts the disturbance in the storage phase. Once micturition has started, these inhibiting reflexes are suppressed. A detrusor contraction that does not result in complete micturition similar to an unstable detrusor contraction could be simulated in a model including urethral afferents. Owing to the number of uncertainties in these models, so far no unambiguous explanation of normal and pathological lower urinary tract behavior can be given. However, these models can be used as an additional tool in studies of the mechanisms of the involved neural control. PMID- 10797585 TI - Preliminary results of myoblast injection into the urethra and bladder wall: a possible method for the treatment of stress urinary incontinence and impaired detrusor contractility. AB - The purpose of this study is to explore the feasibility of myoblasts, the precursors of muscle fibers, injected periurethrally as a potential treatment of stress urinary incontinence. We also studied myoblast injection into the bladder wall to potentially improve detrusor contractility. A myoblast cell line was transduced with adenovirus carrying the expression of the beta-galactosidase reporter gene while in culture. The cells were incubated with fluorescent latex microspheres (FLMs) to follow the outcome of the injected cells. The tissue was harvested 3-4 days after injection; sectioned, fixed, assayed for beta galactosidase expression, and counterstained with H+E. Photographs of the slides were taken under light and fluorescence microscopy. We have noted a large number of cells expressing beta-galactosidase and containing FLMs in the urethral and bladder walls under fluorescent microscopy (8 animals). Many regenerative myofibers expressing beta-galactosidase were also seen in the urethral and bladder walls. The fusion of injected myoblasts to form myotubes was seen in both the urethral and bladder walls. The introduction of myoblasts into the urethral and bladder wall is feasible and results in formation of myotubes and myofibers in the smooth muscle layers of the lower urinary tract. We hypothesize that myoblast injections can be used as a non-allergenic agent to enhance urethral closure and bladder function. PMID- 10797587 TI - Neuroregeneration and voiding behavior patterns after pudendal nerve crush in female rats. AB - Since the pudendal nerve innervates the external urethral sphincter, pudendal nerve injury and resultant neuroregeneration should affect voiding behavior. In this study, neuroregenerative activity of pudendal nerve was correlated to the changes in urinary behavior in female rats. Eighteen female rats underwent bilateral pudendal nerve crush, and 17 to 21 age-matched rats were used as unoperated controls. Urinary volume and frequency were recorded 6 and 13 days post-operatively (dpo). Initiation of pudendal nerve regeneration was indicated by an upregulation of beta(II) tubulin mRNA in the dorsolateral motoneurons (DLM), as measured at 7 and 14 dpo by in situ hybridization with radio-labeled beta(II) tubulin cDNA. At 6 dpo, mean volume voided by the crush group was significantly decreased compared to the control group during the light cycle (P < 0.05). At 7 dpo, the DLM mRNA level was significantly increased in the nerve crush group compared to the control group (P < 0.05). At 13 dpo, there were no differences in volume or frequency between the two groups, suggesting a return to normal voiding behavior. At 14 dpo, there was no significant difference in DLM mRNA levels between crush and control groups. Initiation of nerve regeneration occurs before normalization of voiding behavior after pudendal nerve crush. This data suggest that treatments to accelerate nerve regeneration would improve functional recovery of neurologically based incontinence. PMID- 10797588 TI - James Van Gundia Neel. PMID- 10797589 TI - Resolution passed by the IGES board of directors: james V. Neel, A visionary leader among leaders PMID- 10797590 TI - Score tests for familial correlation in genotyped-proband designs. AB - In the genotyped-proband design, a proband is selected based on an observed phenotype, the genotype of the proband is observed, and then the phenotypes of all first-degree relatives are obtained. The genotypes of these first-degree relatives are not observed. Gail et al. [(1999) Genet Epidemiol] discuss likelihood analysis of this design under the assumption that the phenotypes are conditionally independent of one another given the observed and unobserved genotypes. Li and Thompson [(1997) Biometrics 53:282-293] give an example where this assumption is suspect, thus suggesting that it is important to develop tests for conditional independence. In this paper, we develop a score test for the conditional independence assumption in models that might include covariates or observation of genotypes for some of the first degree relatives. The problem can be cast more generally as one of score testing in the presence of missing covariates. A standard analysis would require specifying a distribution for the covariates, which is not convenient and could lead to a lack of model-robustness. We show that by considering a natural conditional likelihood, and basing the score test on it, a simple analysis results. The methods are applied to a study of the penetrance for breast cancer of BRCA1 and BRCA2 mutations among Ashkenazi Jews. PMID- 10797591 TI - Correcting for ascertainment bias of relative-risk estimates obtained using affected-sib-pair linkage data. AB - Locus-specific sibling relative risk is often estimated using affected-sib-pair lod score analysis of affected sibships and may be used to decide whether to continue or discontinue the search for additional susceptibility genes. We showed that relative-risk estimates obtained using affected-sib-pair data are asymptotically unbiased when each pair is given a weight inversely proportional to the sibship ascertainment probability. Here we show by simulation that the extent of the bias of relative risks estimated using the incorrect ascertainment weights is small for dominant models, but large for single-locus recessive models and some two-locus heterogeneity models. Since in practice the ascertainment scheme is often unknown, we investigate methods for jointly estimating ascertainment and relative risks from affected-sibship data. Given a sufficient sample size, a reasonable estimate of relative risk may always be obtained using only affected pairs from sibships with two affected and no unaffected siblings. This estimate, which has a large variance, may then be used in a three-stage procedure (which we call the alpha method) to estimate consistently both the ascertainment probabilities and the relative risks with greater precision. We additionally propose correction factors to eliminate small-sample bias of relative risks and investigate the bias due to error in the estimate of disease locus location. PMID- 10797592 TI - Power comparison of regression methods to test quantitative traits for association and linkage. AB - George et al. [1999 Am J Hum Genet 65:236-245] proposed a regression-based TDT method for quantitative traits consisting of regressing the trait on the parental transmission of a marker allele. Zhu and Elston [2000] also developed a TDT method for quantitative traits by defining a linear transformation to condition out founder information. Both methods test the null hypothesis of no linkage or association and can be applied to general pedigree structures. In this paper, we compare the power of these two methods through simulation, sampling those nuclear families with at least one heterozygous parent. Overall, we find that a variant of Zhu and Elston's method with 2 d.f. is more powerful. However, if the mode of inheritance is known, then a most powerful test with 1 d.f. can be found. All these regression TDT tests require linkage to detect association, but a test that does not require linkage will be more powerful. PMID- 10797594 TI - Framework for identifying quantitative trait loci in association studies using structural equation modeling. AB - In this article, we suggest a framework for identifying quantitative trait loci (QTL) in association studies using structural equation modeling. Two tests to detect QTLs and estimate the proportion of variance they explain are discussed. The first test assumes that there is no population admixture and only requires that the subjects are genotyped. The second one is a TDT-like test that cannot give false-positive results due to population admixture but requires that the parents of the subjects are genotyped as well and that subjects have at least one heterozygous parent. Power calculations showed that with the first test, 100 subjects were generally sufficient to detect a locus that explained 10% and less than 1,000 subject to detect a locus that explained 1% of the total variance. To obtain the same power, the TDT-like test required an initial sample that was on average 1.7 times larger. Calculations showed that the first test was quite robust against population admixture and that the power of tests to detect admixture was good. This suggested that in the extreme and very specific conditions in which population admixture may cause false-positive findings, admixture can often be detected. PMID- 10797593 TI - ApoE polymorphism accounts for only part of the genetic variation in quantitative ApoE levels. AB - ApoE levels and chromosome 19 ApoE polymorphisms were measured in a sample of 156 Dutch families. Each pedigree consisted of parents aged 35-65 years and their twin offspring aged 14-21 years. A significant effect of the chromosome 19 apoE locus on quantitative plasma levels of apolipoprotein E was observed. The ApoE polymorphism explained 16% of the variance in ApoE levels. Tests of association of ApoE levels with the apoC1 locus, which is in complete linkage disequilibrium with the ApoE locus, also showed a significant effect, although the variance explained by ApoC1 was only 1%. Examination of the covariance between twins classified according to allele sharing indicates that the association is not due to population stratification, but to a genuine effect of the ApoE locus on levels. However, the ApoE locus accounts for only one-fourth of the genetic variation in ApoE levels. PMID- 10797595 TI - Genetic inheritance of body mass index in African-American and African families. AB - Numerous studies have shown recessive major gene inheritance of body mass index (BMI) in white populations; few have examined the inheritance of BMI in the African-American population where obesity is more prevalent, nor in African populations where obesity is comparatively rare. To evaluate the inheritance of obesity in two different populations of African origin, we used segregation analysis to determine the transmission of BMI in 95 African-American families and 400 Nigerian families. Probands were selected from participants in the population based International Collaborative Study on Hypertension in Blacks. Using class D regressive models, results from the segregation analysis of the African-American data showed evidence of a major gene effect on BMI. The Nigerian results were strikingly similar, with comparable estimates for the genotype frequencies and means and strong evidence for a major effect in the transmission of BMI. The high BMI allele frequency estimate of 24% is consistent with estimates in other studies, but the mode of transmission appeared codominant, which differs from studies involving predominantly white populations. In the Nigerian analysis, however, the probability of a high BMI homozygote parent transmitting a low BMI allele to his/her offspring was significantly different from the Mendelian expectation of zero (estimated tau(BB) = 0.45), suggesting that additional complexities exist in the major gene inheritance of BMI in this population. The strong similarity of the genotype frequencies and means obtained from the African American and Nigerian samples suggests that a common codominant major gene effect may contribute to the variation in BMI in both populations. PMID- 10797596 TI - Insights into the structural and functional evolution of plant genomes afforded by the nucleotide sequences of chromosomes 2 and 4 of Arabidopsis thaliana. AB - The rapidly accumulating genome sequence data from the plant Arabidopsis thaliana allows more detailed analysis of genome content and organisation than ever before possible in plants. The genome shows a surprisingly high level of genetic redundancy, with as many as 75% of gene products showing significant homology to another protein of A. thaliana. Many duplicated genes occur in arrays of conserved order and indicate that A. thaliana is likely to have had a tetraploid ancestor. Analysis of the divergence of duplicated genome segments leads to the prediction of two major modes of plant genome evolution: macro-scale duplication and rearrangement of chromosomes and micro-scale translocations, duplication and loss of individual genes or small groups of genes. PMID- 10797597 TI - Graphical tools for comparative genome analysis. AB - Visualization of data is important for many data-rich disciplines. In biology, where data sets are becoming larger and more complex, graphical analysis is felt to be ever more pertinent. Although some patterns and trends in data sets may only be determined by sophisticated computational analysis, viewing data by eye can provide us with an extraordinary amount of information in an instant. Recent advances in bioinformatic technologies allow us to link graphical tools to data sources with ease, so we can visualize our data sets dynamically. Here, an overview of graphical software tools for comparative genome analysis is given, showing that a range of simple tools can provide us with a powerful view of the differences and similarities between genomes. PMID- 10797598 TI - Pharmacogenomics: the genomics of drug response. AB - Pharmacogenomics is defined as the study of the association between genetics and drug response. This is a rapidly expanding field with the hope that, within a few years, prospective genotyping will lead to patients being prescribed drugs which are both safer and more effective ('the right drug for the right patient', or personalized medicine). There are many existing examples in the literature of strong associations between genetic variation and drug response, and some of these even form the basis of accepted clinical tests. The molecular basis for some of these associations is described, and includes examples of variation in genes responsible for absorption and metabolism of the drug, and in target and disease genes. However, there are many issues surrounding the legal, regulatory and ethical framework to these studies that remain unanswered, and a huge amount of education both for the public and healthcare professionals will be needed before the results of this new medicine can be widely accepted. PMID- 10797600 TI - Caenorhabditis elegans. AB - Transparent, free-living nematode worm. Unsegmented body plan with full set of differentiated tissues (neural, endoderm, ectoderm and muscle). Genome size approximately 97 Mb, as five autosomes and one X sex chromosome. Fully sequenced genome, which comprises approximately 20 000 predicted genes. Defined cell lineage. Has made major contribution to studies of development, cell-to-cell signalling, cell ageing and cell death processes. Large-scale gene deletion, microarray analysis of gene expression and two-hybrid protein interaction analysis projects under way. Comparative studies mainly with C. briggsae, but also with other free-living and parasitic nematodes. PMID- 10797599 TI - Estimation of synteny conservation and genome compaction between pufferfish (Fugu) and human. AB - BACKGROUND: Knowledge of the amount of gene order and synteny conservation between two species gives insights to the extent and mechanisms of divergence. The vertebrate Fugu rubripes (pufferfish) has a small genome with little repetitive sequence which makes it attractive as a model genome. Genome compaction and synteny conservation between human and Fugu were studied using data from public databases. METHODS: Intron length and map positions of human and Fugu orthologues were compared to analyse relative genome compaction and synteny conservation respectively. The divergence of these two genomes by genome rearrangement was simulated and the results were compared to the real data. RESULTS: Analysis of 199 introns in 22 orthologous genes showed an eight-fold average size reduction in Fugu, consistent with the ratio of total genome sizes. There was no consistent pattern relating the size reduction in individual introns or genes to gene base composition in either species. For genes that are neighbours in Fugu (genes from the same cosmid or GenBank entry), 40-50% have conserved synteny with a human chromosome. This figure may be underestimated by as much as two-fold, due to problems caused by incomplete human genome sequence data and the existence of dispersed gene families. Some genes that are neighbours in Fugu have human orthologues that are several megabases and tens of genes apart. This is probably caused by small inversions or other intrachromosomal rearrangements. CONCLUSIONS: Comparison of observed data to computer simulations suggests that 4000-16 000 chromosomal rearrangements have occurred since Fugu and human shared a common ancestor, implying a faster rate of rearrangement than seen in human/mouse comparisons. PMID- 10797601 TI - Nematode functional genomics. PMID- 10797605 TI - Correction for age of anticardiolipin antibodies cut-off points. AB - Immunoglobulin (Ig) isotypes G, M, and A of anticardiolipin antibodies (aCL) are considered markers of antiphospholipid syndrome (APS). They were determined by ELISA in sera of 100 healthy children aged between 6 months and 16 yrs (mean 5.4 yrs +/-3.4 SD) and 100 healthy elderly subjects aged between 65 and 103 yrs (mean 84.2 yrs +/-8.1 SD). The frequency with which they were detected was compared to that in sera of 100 healthy adults aged between 21 and 47 yrs (mean 25.8 yrs +/ 5.2 SD) in order to evaluate if adult aCL cut-off levels were fit for pediatric and elderly populations. The cut-off points were calculated adding 2.5 SD to the mean values and, in the adult group, the results were 11.6 GPL, 7.5 MPL, and 23.9 APL for IgG, IgM, and IgA, respectively. In the children, IgG aCL were positive in 26 cases (26%), IgM and IgA aCL in 1 case (1%) respectively. Statistical comparison of these results to those of adults showed a higher significant frequency for children IgG aCL (P = 0.0001) with the major contribution by children aged between 6 months and 5 yrs, and a lower significant frequency for children IgA aCL (P = 0.041). In elderly subjects IgG aCL were positive in 12 cases (12%), IgM aCL in 4 (4%), and IgA aCL in 37 (37%). In a comparison of these values to those of adults, only elderly IgA aCL frequency was significantly higher (P = 0.0001) with the major contribution by the oldest subgroup. In order to avoid false positive results for IgG aCL in children and for IgA aCL in elderly, as well as false negative results for IgA aCL in children, we introduced three new cut-off points: (1) 27.7 GPL for IgG aCL in children; (2) 13.8 APL for IgA aCL in children; and (3) 51.1 APL for IgA aCL in elderly subjects. These data suggest that a correction for age of aCL cut-off levels should be considered to raise specificity and sensitivity for APS in pediatric as well as in elderly populations. PMID- 10797607 TI - Availability of Nagarse for DNA analysis as a substitute for proteinase K. AB - The availability of Nagarse, a protease, as a substitute for proteinase K for digestion of leukocytic or bacterial DNAs was studied. The amount and purity of DNAs extracted from leukocytes and several bacterial strains with Nagarse were compared with those of DNAs treated with proteinase. Nagarse exhibited the same behavior as proteinase K in digesting leukocytes, and it could also be used for bacterial digestion for physical mapping of genomic DNA by biased sinusoid field gel electrophoresis. Nagarse was thus comparable to proteinase K for use in biochemical experiments. The principal advantage of Nagarse is that it is inexpensive, unlike proteinase K, and our findings indicated that Nagarse is very useful as a substitute for proteinase K for the DNA study. PMID- 10797606 TI - Influence of cholecalciferol (vitamin D3) on the course of experimental systemic lupus erythematosus in F1 (NZBxW) mice. AB - The course of systemic lupus erythematosus (SLE), an autoimmune disease, is markedly affected by hormones such as estrogen and prolactin. It is well known that heavy exposure to sunlight has deleterious effects on SLE, triggering episodes of the disease. Classical explanations for this occurrence suggest that UV radiation damages DNA, which becomes immunogenic, or induces exposure of the Ro antigen in keratinocytes. In recent years, it has been shown that vitamin D3 has important effects on the immune system. Thus, we proposed an alternative hypothesis, suggesting that UV radiation, by promoting vitamin D3 synthesis, could be a factor aggravating the course of SLE after exposure to sunlight. To test this hypothesis, we injected F1(NZBxW) mice, which are prone to developing SLE, with vitamin D3, and we demonstrated a worsening of the histopathological findings in the kidney. PMID- 10797602 TI - Current awareness on comparative and functional genomics [bibliography]. AB - In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on comparative and functional genomics. Each bibliography is divided into 16 sections. 1 Reviews & symposia; 2 General; 3 Large-scale sequencing and mapping; 4 Genome evolution; 5 Comparative genomics; 6 Gene families and regulons; 7 Pharmacogenomics; 8 Large-scale mutagenesis programmes; 9 Functional complementation; 10 Transcriptomics; 11 Proteomics; 12 Protein structural genomics; 13 Metabolomics; 14 Genomic approaches to development; 15 Technological advances; 16 Bioinformatics. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted PMID- 10797608 TI - Clinical utilization of the international normalized ratio (INR). AB - The prothrombin time (PT) is one of the most important laboratory tests to determine the functionality of the blood coagulation system. It is used in patient care to diagnose diseases of coagulation, assess the risk of bleeding in patients undergoing operative procedures, monitor patients being treated with oral anticoagulant (coumadin) therapy, and evaluate liver function. The PT is performed by measuring the clotting time of platelet-poor plasma after the addition of calcium and thromboplastin, a combination of tissue factor and phospholipid. Intra- and interlaboratory variation in the PT was a significant problem for clinical laboratories in the past, when crude extracts of rabbit brain or human placenta were the only source of thromboplastin. The international normalized ratio (INR), developed by the World Health Organization in the early 1980s, is designed to eliminate problems in oral anticoagulant therapy caused by variability in the sensitivity of different commercial sources and different lots of thromboplastin to blood coagulation factor VII. The INR is used worldwide by most laboratories performing oral anticoagulation monitoring, and is routinely incorporated into dosage planning for patients receiving warfarin. Although the recent availability of sensitive PT reagents prepared from recombinant human tissue factor (rHTF) and synthetic phospholipids eliminated many of the earlier problems associated with the use of crude thromboplastin preparations, local instrument variability in the INR still remains a problem. Presently, the use of plasma calibrants seems the best solution to this problem. Standardizing the point-of-care instruments for INR monitoring is another dilemma faced by the industry. Ultimately, new generations of anticoagulant drugs may eliminate the need for laboratory monitoring of anticoagulant therapy. PMID- 10797609 TI - Utilization of spectral absorption for measurement of adenylate cyclase activity. AB - The purpose of this study was to improve our previously described enzymatic fluorometric assay of the adenylate cyclase activity. Using physicochemical characteristics of NADPH, of which a 0.1 mmol/L solution would have an optical density of 0.627, we measured the adenylate cyclase activity by the spectral absorption of NADPH. The assay consists of two parts: pharmacological modulation of adenylate cyclase and measurement of newly synthesized cyclic AMP. The latter part involves four steps: enzymatic destruction of noncyclic adenine nucleotides and phosphorylated metabolites, conversion of cyclic AMP to ATP, amplification of ATP by enzymatic cycling, and measurement of NADPH with spectral absorption, which is generated in proportion to initial cyclic AMP levels. This new assay was tested in membrane preparations made from rat hearts in comparison with the previously described fluorometric assay. We obtained identical results by spectrophotometry and fluorometry with high reproducibility. Because the fluorometric assay possesses a high sensitivity while the spectrophotometric method is advantageous because of its wide analytical range of cyclic AMP measurement, combination of fluorometric and spectrophotometric methods may offer a convenient way to measure the adenylate cyclase activities in various samples. PMID- 10797610 TI - Decrease of HCVRNA after three days of daily interferon treatment is predictive of the virological response at one month. AB - Early monitoring of HCVRNA during interferon treatment may allow clinicians to obtain important information that could help them to adopt therapeutic decisions in individual cases. The hepatitis C virus infection is highly dynamic and a daily high dose of IFN may induce a decline of viremia of 95+/-10% of baseline value after 24 to 48 hours of treatment. The importance of this early antiviral efficacy has not been understood. We have measured HCVRNA levels in 47 patients with chronic hepatitis C infection during interferon treatment to study HCVRNA kinetics and evaluate the predictive value of the early decay of viremia on the virological response after one month of treatment. Sixty percent of treated patients showed early virological response (EVR) and it was significantly associated with low HCVRNA levels and a genotype other than 1b. Finally, the absence of an 85% decline in HCVRNA levels after 3 days of treatment observed in 11 out of 45 patients (24%) was an absolute and very early predictor of the absence of EVR in the study population. PMID- 10797611 TI - Lipid-lowering response of the HMG-CoA reductase inhibitor fluvastatin is influenced by polymorphisms in the low-density lipoprotein receptor gene in Brazilian patients with primary hypercholesterolemia. AB - Although the efficacy of fluvastatin (HMG-CoA reductase inhibitor) in the treatment of primary hypercholesterolemia is well documented, a wide interindividual variation treatment response has been observed. We have studied the possible role of the AvaII (exon 13), HincII (exon 12), and PvuII (intron 15) polymorphisms at the low-density lipoprotein receptor (LDLR) gene on lipid lowering response in 55 patients (36 to 70 years old) with primary hypercholesterolemia treated with fluvastatin for 16 weeks. LDLR genotypes were determined by PCR-RFLP. The results indicate that the AvaII and PvuII polymorphisms influence the cholesterol-lowering response of the HMG-CoA reductase inhibitor Fluvastatin. Patients carrying A+A+ (AvaII) or P1P1 (PvuII) homozygous genotypes presented lower reduction in total cholesterol, LDL-C and apolipoprotein B levels after 16 weeks of treatment with fluvastatin, when compared to other genotypes (P<0.05). Our data also support the previous assumption that the AvaII, HincII, and PvuII polymorphisms of the LDLR gene are associated with variation of serum cholesterol levels. Therefore, the identification of the LDLR genetic profile may provide better prediction of a patient's clinical response to fluvastatin. PMID- 10797612 TI - Effects of galactolipid elimination on oligodendrocyte development and myelination. AB - The galactolipids galactocerebroside and sulfatide, which require the enzyme UDP galactose:ceramide galactosyltransferase (CGT) for their synthesis, are among the most prevalent molecules in the myelin sheath. Numerous studies, mainly using antibody perturbation methods in vitro, have suggested that these molecules are crucial mediators of oligodendrocyte differentiation and myelin formation. Although we have previously demonstrated that myelin formation occurs in CGT null mutant mice, which are incapable of synthesizing the myelin galactolipids, here we show that there are developmental alterations in the CNS of these animals. There is a significant decrease in the number of myelinated axon segments in the mutant spinal cord despite normal levels of myelin gene-specific mRNAs and proteins. Also, there is an increased cellularity in the mature mutant spinal cord and the distinctive morphology of the additional cells suggests that they are actively myelinating oligodendrocytes. Using in situ hybridization techniques, we show that there is a 50% increase in the number of oligodendrocytes in the mutant spinal cord. The data suggest that galactolipids play an important developmental role in regulating the maturation program and final number of oligodendrocytes. PMID- 10797613 TI - Evaluation of glutathione-sensitive fluorescent dyes in cortical culture. AB - The sensitivity of six fluorophores to glutathione (GSH) was evaluated in living rat cortical neuronal/glial mixed cultures during the first 23 days in vitro (DIV). Four of the dyes require glutathione-S-transferase (GST) to form a fluorescent conjugate, potentially conferring specificity for GSH: these included t-butoxycarbonyl-Leu-Met-7-amino-4-chloromethylcoumarin (CMAC), 7-amino-4 chloromethylcoumarin (CMAC-blue), monochlorobimane (MCB), and 5 chloromethylfluorescein diacetate (CMFDA). The final two dyes examined, 2,3 naphthalenedicarboxaldehyde (NDA) and o-phthaldehyde (OPD), do not require GST for adduct formation with GSH. To examine the specificity of the dyes for GSH, cultures grown less than 6 DIV were pretreated with diethyl maleate or DL buthionine-(S, R)-sulfoximine to deplete endogenous GSH. This resulted in a substantial loss of staining by CMAC, CMAC-blue, and MCB and partial loss of staining by OPD, indicating specificity for GSH, while staining by CMFDA or NDA was not altered, indicating a lack of specificity for GSH. Neurons experienced a dramatic decline in GSH levels relative to astrocytes between 5-6 DIV, as shown by a loss of neuronal staining with CMAC, CMAC-blue and MCB. This decrease in staining was not due to a decrease in GST activity, as neurons stained with the GST-insensitive OPD also exhibited a decline in GSH-sensitive staining. Immunolabeling experiments demonstrated that CMAC staining co-localized with GFAP positive astrocytes, but not with MAP-2-positive neurons, in 18 DIV cultures. Finally, CMAC was exploited as a specific morphological marker of astrocytes in cultures aged >5 DIV. CMAC staining was employed to monitor astrocyte proliferation and to resolve astrocytes in living mixed cultures co-loaded with the Ca(2+)-sensitive dye, calcium green 5N-AM. GLIA 30:329-341, 2000. Published 2000 Wiley-Liss, Inc. PMID- 10797614 TI - Temporal analysis of growth factor mRNA expression in myelinating rat brain aggregate cultures: increments in CNTF, FGF-2, IGF-I, and PDGF-AA mRNA are induced by antibody-mediated demyelination. AB - Myelinogenesis in rat brain aggregate cultures is associated with a pattern of growth factor mRNA expression comparable to that of the developing brain. The rate of increase in platelet-derived growth factor-AA (PDGF-AA) expression was greatest just before the detection of myelin basic protein (MBP) mRNA in the cultures and remained high thereafter, consistent with in vivo observations. Levels of fibroblast growth factor-2 (FGF-2) and of ciliary neurotrophic factor (CNTF) mRNA increased continuously over the period of MBP accumulation. High rates of transforming growth factor beta1 (TGF-beta1), insulin-like growth factor I (IGF-I), and neurotrophin-3 (NT-3) expression at early time points during the culture gradually decreased over time, indicative of a key regulatory role during oligodendrocyte development. The addition of demyelinative anti-myelin oligodendrocyte glycoprotein (anti-MOG) antibody resulted in a significant increase in MBP peptide fragments with a C-terminus at phenylalanine 89 indicating proteolytic breakdown of MBP after myelin phagocytosis. Immediately after antibody treatment the expression of CNTF mRNA was significantly increased, compared with controls, while that of FGF-2 and IGF-I, and of PDGF-AA peaked during the early and later stages of recovery respectively. Thus, specific growth factors combine to regulate myelination and remyelination in the aggregates; these data have implications for demyelinating disease in which protective growth factor secretion may be central to regeneration. PMID- 10797615 TI - Expression of metabotropic glutamate receptor 5 is increased in astrocytes after kainate-induced epileptic seizures. AB - In the CNS there is a differential distribution of the metabotropic glutamate receptor 5 (mGluR5) in neurons and glia. Hippocampal nerve cells contain large amounts of the receptor transcript and protein that are expressed at very low levels in astrocytes. This is unexpected, as mGluR-induced phosphoinositide hydrolysis is substantial in cultured astrocytes and is mediated only by mGluR5 in these cells. In order to detect mGluR5 in astrocytes in vivo, we destroyed in a circumscribed part of the hippocampus nerve cells that have high level of receptor expression. Unilateral injection of kainate into the right amygdala produced epileptic seizures, as well as selective degeneration of nerve cells restricted to the ipsilateral CA3 and CA4 regions of the hippocampus, followed by the development of gliosis. In the affected fields only, mGluR5 immunoreactivity was severely reduced 3 days after kainate injection, followed by a progressive reappearance and lasting presence of the receptor protein. Receptor mRNA virtually disappeared from the pyramidal cell layer of the lesioned CA3/4 region. On the other hand, the message level increased persistently in the CA3 stratum lucidum and radiatum, the site of massive astrogliosis. In these sites, mGluR5 mRNA became detectable in double labeling studies in glial fibrillary acidic protein-positive astrocytes. We showed previously that growth factors induce a pronounced elevation of mGluR5 expression in cultured astrocytes (Miller et al. J Neurosci 15:-6109, 1995). The well-documented increase in the level of growth factors in the damaged brain may underlie the induction of the receptor expression in astrocytes in vivo, which may also be involved in repair processes in the injured nervous tissue. PMID- 10797616 TI - Glial heterogeneity in expression of the inwardly rectifying K(+) channel, Kir4.1, in adult rat CNS. AB - Previous electrophysiological evidence has indicated that astrocytes and oligodendrocytes express inwardly rectifying K(+) channels both in vitro and in vivo. Here, for the first time, we have undertaken light microscopic immunohistochemical studies demonstrating the location of one such channel, Kir4.1, in both cell types in regions of the rat CNS. Some astrocytes such as those in the deep cerebellar nuclei, Bergmann glia, retinal Muller cells, and a subset in hippocampus express Kir4.1 immunoreactivity, but not others including those in white matter. Oligodendrocytes also express this protein, strongly in perikarya and to a lesser extent in their processes. Expression of Kir4.1 in astrocytes and oligodendrocytes would enable these cells to clear extracellular K(+) through this channel, whereas nonexpressors might use other mechanisms. PMID- 10797617 TI - Regulation of Fas and FasL expression on rat Schwann cells. AB - Although the PNS belongs to the immune privileged sites, it can become a target of immune attacks by invading T cells, causing inflammation and destruction. Yet the PNS also has a protective potential by eliminating the inflammatory cells via apoptosis. In analogy with other immune-protected sites, participation of the apoptosis-inducing Fas/FasL molecules could play an important role. To assess the possible involvement of the Fas/FasL system in T-cell apoptosis in the PNS of the rat, we characterized Fas and FasL expression on neonatal rat Schwann cells (SC) in vitro. Cells were stimulated in vitro with interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), or a combination of both. We observed upregulation of FasL expression under the influence of IFN-gamma, while adding TNF-alpha alone to the culture medium had no effect. IFN-gamma and TNF-alpha acted synergistically, leading to an increased FasL expression that reached its maximum 70 h after cytokine exposure, as shown by FACS analysis, SDS-PAGE, and Western blot. Fas expression on untreated SC showed fluctuating levels, while addition of IFN-gamma and TNF-alpha suppressed Fas expression completely. These findings are in accord with recently published results showing Fas and FasL expression on malignant human cells, derived from brain tumors and upregulation of FasL on astrocytes after exposure to IFN-gamma and TNF-alpha. Furthermore, FasL-expressing SC could be revealed by immunostaining of sciatic nerve from Lewis rats suffering from experimental autoimmune neuritis (EAN). We suggest that Fas/FasL expression on SC may contribute to the elimination of invading autoreactive T cells in the PNS in concert with other immune defense mechanisms. PMID- 10797618 TI - Protein targeting to glycogen mRNA expression is stimulated by noradrenaline in mouse cortical astrocytes. AB - Brain glycogen levels are dynamically regulated by certain neurotransmitters, including noradrenaline (NA) and vasoactive intestinal peptide (VIP). In particular, glycogen synthesis involves activation by NA and VIP of the transcription factors C/EBPbeta and -delta as well as the induction of glycogen synthase. Glycogen accumulation is found in a variety of neuropathological conditions, including reactive astrocytosis after CNS lesions, as well as in Alzheimer's disease. Protein targeting to glycogen (PTG) belongs to a family of proteins that play a key role in glycogen synthesis in peripheral tissues. In this study, we report the presence of PTG mRNA in adult mouse brain, as well as in astrocytes, a non-neuronal cell type that contains most of brain glycogen. Using primary cultures of mouse cortical astrocytes, we observed that NA leads to time- and concentration-dependent induction of PTG mRNA expression. This effect, concomitant to an enhancement of glycogen synthesis in these cells, depends on the activation of beta(1)-adrenergic receptors. Induction of PTG mRNA expression was mimicked by the adenylate cyclase activator forskolin and by dibutyryl cAMP, suggesting the involvement of the cAMP-dependent signal transduction cascade. Among other neuroactive substances known to elevate glycogen levels in astrocytes, VIP had a comparable effect to that of noradrenaline, whereas insulin and glutamate were without effect on PTG mRNA expression. These data suggest that increased PTG expression by neurotransmitters such as noradrenaline and VIP could represent a major event leading to enhancement of glycogen levels in astrocytes. PMID- 10797619 TI - Enhancement of glutamate uptake transport by CO(2)/bicarbonate in the leech giant glial cell. AB - Glutamate uptake into glial cells via the excitatory amino acid transporter (EAAT) is accompanied by an influx of sodium and acid equivalents into the cells. The sodium-bicarbonate cotransport (NBC) in glial cells moves sodium and base equivalents across the glial membrane in both directions. We have studied possible interactions between these two electrogenic transporters in the giant glial cell of isolated ganglia of the leech Hirudo medicinalis. Changes in membrane potential, membrane current, intracellular sodium, and intracellular pH evoked by aspartate (1 mM), an EAAT agonist, were measured both in the absence and in the presence of CO(2)/bicarbonate. When 5% CO(2) and 24 mM bicarbonate was added to the saline (at constant pH 7.4), the aspartate-induced membrane current was increased, while the change in intracellular sodium was decreased. The acid influx evoked by aspartate was enhanced by CO(2)/bicarbonate but, because of the increased intracellular CO(2)/bicarbonate-dependent buffering power, the change in intracellular pH was decreased. 4,4'-Diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS, 0.5 mM), which inhibits the NBC, reversed the effects of CO(2)/bicarbonate on the aspartate-induced current and pH change. Our results suggest that the NBC helps counteract dissipation of the sodium and the acid-base gradients induced by the EAAT, enhancing the rate and capacity of glutamate uptake by glial cells. PMID- 10797620 TI - Differential regulation of microglial keratan sulfate immunoreactivity by proinflammatory cytokines and colony-stimulating factors. AB - Resident microglia of the rat CNS express a unique type of keratan sulfate immunoreactivity (KS-IR) that is lacking on peripheral monocytes/macrophages and associated with a so far unknown proteoglycan core protein. Microglial KS-IR is downregulated during T-cell-mediated autoimmune inflammation but largely preserved in degenerative lesion paradigms. This study addresses the role of cytokines and colony-stimulating factors in the regulation of microglial KS-IR. In vitro, ramified microglia in coculture with astrocytes, but not isolated microglia, constitutively expressed KS-IR under control conditions. In both culture paradigms, KS-IR was increased significantly by macrophage- (M-CSF) and granulocyte/macrophage colony-stimulating factors (GM-CSF), as well as tumor necrosis factor-alpha (TNF-alpha). By contrast, the Th1 cytokine interferon-gamma (IFN-gamma) downregulated KS-IR, both when applied alone or in combination with either GM-CSF, M-CSF, or TNF-alpha. In vivo, the intracerebroventricular administration of IFN-gamma, but not TNF-alpha, to healthy rats led to an almost complete disappearance of KS-IR from ramified brain microglia. Our data suggest that the expression of microglial KS-IR is under dominant negative control by the Th1 cell cytokine IFN-gamma and represent the first evidence of cytokine dependent proteoglycan regulation in the CNS. PMID- 10797621 TI - Morphometric study of the sphincter of oddi (hepatopancreatic) and configuration of the submucosal portion of the sphincteric muscle mass. AB - The sphincter of Oddi (SO) hepatopancreatic sphincter from 114 Japanese adults, especially the sphincteric muscle mass lying in the duodenal mucosal layer (the submucosal portion of the SO: SMSO), was measured macroscopically under a binocular microscope. The SMSO was classified into two types according to shape. The horizontal type (95/114) usually displayed a rod-like shape (6.0 mm at average diameter at the root) directed toward the anus and was situated on the duodenal muscle wall with the papillary orifice at or near the tip (anal side) of the SMSO. The horizontal type SMSO followed the underlying muscle wall widely ranging from 5.0 mm to 17.9 mm length (9.8 mm at average) and was attached to the wall by the loose connective tissue along the entire length. In the horizontal type, the extramural portion (the portion penetrating and outside of duodenal muscle of the SO) of the SO was very small. The vertical type (19/114) erected on the muscle wall vertically into the lumen of the duodenum. The average length and average maximum diameter of the vertical type were 6.0 mm and 6.2 mm, respectively. The extramural portion of the SO was thicker and tighter in the vertical type, which suggested that endoscopic sphincterotomy might preserve the length of the SO longer than endoscopic papillary balloon dilation (EPBD). Of the individuals in our study, 18% (19/114) had SO diameters <5 mm, a finding that may have implications for selection of balloon size in EPBD. PMID- 10797622 TI - Is the renal space closed? AB - The aim of this study is to verify the aperture or the closure of the renal space. The study is undertaken on histological sections of fetuses in the horizontal and sagittal planes. On each side, the kidney and the suprarenal gland are disposed in a space that is closed on all sides. The anterior and posterior layers of the renal fascia fuse at the upper pole of the space to become continuous with the inferior fascia of the diaphragm. Likewise, they merge at the lower pole and at the lateral border of the space to become continuous with the fasciae of the parietal muscles. At the medial border of the space, the two layers merge to continue medially with the peri-aortocaval connective tissue; they penetrate the hilum and beneath it enclose the ureter. PMID- 10797623 TI - Gross morphology and ultrastructure of dendritic cells in the normal human decidua. AB - Five normal placentae of normal pregnancy and delivery were used to study the gross morphology and ultrastructure of the dendritic cells in the normal human decidua. Zinc iodide osmium (ZIO) mixture was prepared. Small pieces of the placenta were processed for light microscopy and electron microscopy. For light microscopy, the small pieces of placenta were incubated in 20 mM PBS-EDTA solution, ph 7.4 at 37 degrees C to detach the basal plate. The basal plate pieces were incubated in ZIO. A wholemount preparation of the basal plate demonstrated the whole profile and gross morphology of the dendritic cell. For electron microscopy, the placenta pieces were fixed in 3% glutaraldehyde in 0.1 M phosphate buffer, ph 7.4, washed with phosphate buffer, put in ZIO mixture, washed in distilled water, dehydrated in graded ethanol, cleared in propylene oxide, and embedded in resin. Ultra thin sections of the ZIO blocks were cut using a diamond knife and stained with lead citrate. Ultrastructure of the dendritic cell presented multiple cytoplasmic processes, lobulated or round or oval, heterochromatic or euchromatic nucleus, mitochondria, free ribosomes, and pieces of rough endoplasmic reticulum, but no Birbeck granules. PMID- 10797624 TI - Chilaiditi's syndrome and obesity. AB - Hepatodiaphragmatic interposition (HDI) of the colon or small intestine (Chilaiditi's syndrome) is more prevalent in the elderly and in men. The reason for this is unclear. In obese individuals, fat deposition between the liver and colon widens this potential space, and this may be an important factor in the etiology of this condition. As men store more fat intra-abdominally than women, this may explain the increased prevalence in men. This study was performed to investigate the relationship between Chilaiditi's syndrome and obesity. A total of 850 abdominal CT examinations were assessed for the presence of HDI of the colon or small intestine, and 10 patients with Chilaiditi's syndrome were identified. The present Body Mass Index (BMI) and the highest BMI in the past was recorded for each patient. Eight of the 10 patients were male; five had a BMI greater than 28.5 and three had a BMI between 25 and 27.5 at some period in their lives. Obesity may be a causative factor in the etiology of Chilaiditi's syndrome, and the difference in fat deposition between men and women may explain its increased prevalence in men. PMID- 10797625 TI - Observations on multiple sperm granulomas in the rat epididymis following vasectomy. AB - Seven epididymides of long-term vasectomized rats showing multiple sperm granulomas were studied in serial histological sections. Despite the presence of the multiple granulomas, only two rats showed continuity of the epididymal duct with the central sperm mass of a granuloma. A further three specimens showed breaks in the epididymal epithelium at sites of local distension in the epididymal tail. Granulomas in the epididymal body seem to receive spermatozoa only transiently. The spermatozoa at the center of granulomas in continuity with the epididymal duct showed evenly distributed sperm heads. A number of others showed clumping of spermatozoa, attributed to stagnation of flow and fluid resorption. In many granulomas, folds in the macrophage layer with a connective tissue core rich in lymphocytes and plasma cells projected into the central sperm mass. We conclude that the sperm granuloma is a dynamic structure that shows changes with age and that the sperm granuloma closest to the testis must not be assumed to be the one that is draining the spermatozoa. PMID- 10797626 TI - Incidence of a septum in the first dorsal compartment and its effects on therapy of de Quervain's disease. AB - Because there has been no study of the presence of a septum in the first dorsal compartment of the wrist in Asians and there is no method of conservative treatment of de Quervain's disease based on the presence of this septum, the authors have evaluated this septum in a large Asian population. Two-hundred wrists in 100 Asian cadavers were dissected. The presence of the septum was documented by removing the abductor pollicis longus (APL) tendon from its tunnel in the first dorsal compartment and observing through the same tunnel if there was a septum to prevent immediate access to the extensor pollicis brevis (EPB) tendor. A septum was present in 77.5% of wrists. When the septum was present, the resultant subcompartment for the EPB was quite narrow. Given the high incidence of the septum, therapy of de Quervain's disease could reasonably be modified accordingly. For instance, to improve the success rate for local steroid injection, the solution of steroid should aim at reaching both subjcompartments by redirecting the course of the injecting needle. As for surgical release, adequacy could be enhanced by the realization by the surgeon of the possible presence of the septum. PMID- 10797627 TI - Tibial tubercle position and patellar height as indicators of malalignment in women with anterior knee pain. AB - In this study, we evaluated 70 limbs in 42 women with anterior knee pain. We investigated tibial tubercle position and patellar height indices as indicators of malalignment. Tibial tubercle rotation angles were determined by computed tomography, and patellar height indicators, Insall-Salvati, modified Insall Salvati, Caton, and Blackburne indices were calculated on lateral knee roentgenograms. The results were compared to values obtained from 80 limbs in 40 healthy female volunteers. Tibial tubercle rotation angle was 68.1 degrees (+/ 3.6) in the study group and 70.3 degrees (+/-3.8) in the control group. The difference was statistically significant (P< 0. 01). Patellar height indicators were not statistically different between the two groups. These results suggested that patellar height is not a malalignment indicator in female patients with anterior knee pain. These patients should be investigated by computed tomography to determine tibial tubercle position. PMID- 10797628 TI - Ectopic "high" origin of both coronary arteries from the left aortic wall: anatomic and postmortem angiographic findings. AB - Anatomic and postmortem angiographic findings of a previously unreported case of ectopic origins and unusual courses of the right coronary (RC) artery and the left coronary (LC) artery were demonstrated. This specimen was unique among 450 angiographies and 60 corrosion castings of the human hearts examined in this study. The ostium of the RC artery was pocket-like, located in the left aortic wall at roughly 180 degrees to the long axis of the ascending aorta and 19 mm above the rim of the sinotubular junction (SJ). Initially, the RC runs to the right and downward, passing high in the cleft between the aorta and the pulmonary trunk, thereby avoiding a possible compression from them. The ostium of the LC faced upward and originated from the left aortic wall 7 mm above the SJ. The LC ran to the left and downwards for 16 mm until its division. Histologically, the first 11 mm of the RC were elastic. This observation, together with its high course between the great vessels, combined to make this case benign. The best x ray projections to show the characteristic findings of the present case were anteroposterior and lateral, which were of practical importance for the correct determination and interpretation of this case. The cardiac surgeon should be aware that high cannulation will be required to locate the RC to avoid accidentally cross-clamping or transecting the vessel during surgery where this anomaly may be encountered. PMID- 10797629 TI - Terminology of the prostate and related structures. AB - The various studies of J.E. McNeal (1968ndash;1977) established with precision the different sites of predilection for prostatic carcinomas, benign prostatic hypertrophy (BPH), and inflammation, making the division of the prostate into right, left, and middle lobes and isthmus inadequate for their description. Unfortunately, his positional terms for the sites (peripheral, central, transition, and preprostatic) are not based on the usual parameters and cannot be directly related to those of other workers. Nevertheless, the International Federation of Associations of Anatomists' Federative Committee on Anatomical Terminology has made recommendations in Terminologia Anatomica, based on his findings but modified to take some account of those of Tisell and Salander (1975 Scand J Urol Nephrol 1975 9:185-191). The use of the term lobe is confined to the right and left lobes and the variable middle lobe. The term lobule is used for the subdivisions, which are named from the anatomical position. Thus each side has a superomedial, an anteromedial, an inferoposterior, and an inferolateral lobule. Also necessary to describe a site of predilection is a peri-urethral gland zone. In ultrasound diagnosis, the trapezoid area is important: its upper limit is the rectoperinealis, its anterior limit is the intermediate part of the urethra, its lower limit is the anoperinealis, and its posterior limit is the anorectal junction. Confusion at the bladder neck is resolved by recognizing that the position of the internal urethral orifice varies with functional state of the bladder: while it is filling the orifice lies above the base of the prostate; when voiding begins, the orifice descends to the base of the prostate; between the filling internal orifice and the emptying internal orifice is the bladder neck part of the urethra. PMID- 10797630 TI - Structure of the adult prostate from a clinician's standpoint. AB - When urologists palpate left and right lobes of the prostate on digital rectal examination, they are assessing the gland for the presence of abnormal conditions. Benign prostatic hyperplasia, an abnormal condition may appear lobar by digital examination; however, the normal, nonhyperplastic prostate lacks lobar configuration. PMID- 10797631 TI - Rare variant of the brachial artery: superficial lateral inferior type VII EAB. AB - This study concerns a variant of the brachial artery with a modified origin and course. The artery was studied in 418 upper limbs removed from cadavers. In one upper limb the left brachial artery bifurcated into: (1) a superficial brachial a. (the variant) crossing superficially to the bicipital aponeurosis in the cubital region and assuming the course, position, and supply area of the radial artery, and (2) the deep brachial a. (another variant). An embryologic interpretation of this anomaly is based on a variant vascular development derived from the eighth intersegmental artery. The variant is termed according to Adachi's classification schedule as arteria brachialis superficialis lateralis inferior Type VII with the addition EAB (epiaponeurosis bicipitalis, i.e., superficial to the bicipital aponeurosis). Accurate information concerning unusual patterns of the arteries in the upper limbs is clinically relevant, especially in the avoidance of accidental intra-arterial injection with reflectory vascular occlusion leading to necrosis. PMID- 10797632 TI - Clinical anatomy. PMID- 10797633 TI - Proton magnetic resonance spectroscopy in primary and secondary progressive multiple sclerosis. AB - The objective of this study was to characterize and compare the proton magnetic resonance spectrum of a voxel, containing lesions or normal-appearing white matter (NAWM), in primary-progressive (PP) and secondary-progressive (SP) multiple sclerosis (MS) patients. Single-voxel proton magnetic resonance spectroscopy and magnetic resonance imaging were performed in 35 MS patients (17 PP and 18 SP) and 17 controls. Spectra from an 8 ml voxel located in the parieto occipital region were obtained with a spin-echo pulse sequence (1600 ms/135 ms/256, TR/TE/acquisitions). Resonance areas due to N-acetylaspartate (NAA), creatine/phosphocreatine (Cr) and choline compounds (Cho) were determined, and results expressed in terms of area in arbitrary units or as metabolite ratios. With respect to the control group, there were significant reductions in NAA and NAA/Cho ratio in the PP-lesion, SP-lesion, PP-NAWM and SP-NAWM groups. There were no significant differences between the PP-NAWM and SP-NAWM groups. These results support the existence of metabolic changes in the white matter of PP and SP patients and suggest that there is neuronal damage and/or loss in both clinical courses. Finally, characterization of the parieto-occipital region showed no significant differences in the spectral pattern of NAWM between PP and SP clinical courses of MS. PMID- 10797634 TI - Distinction between normal and renal cell carcinoma kidney cortical biopsy samples using pattern recognition of (1)H magic angle spinning (MAS) NMR spectra. AB - The technique of magic angle spinning (MAS) high resolution (1)H NMR spectroscopy applied to intact tissues provides excellent peak resolution and thus much biochemical information. The use of computer-based pattern recognition techniques to classify human renal cortex tissue samples as normal or tumour based on their (1)H MAS NMR spectra has been investigated. In this preliminary study of 22 paired control and tumour samples, exploratory data analysis using principal components based on NMR spectral intensities showed clear separation of the two classes. Furthermore, using the supervised method of linear discriminant analysis, based on individual data point intensities or on integrated spectral regions, it was possible to distinguish between the normal and tumour kidney cortex tissue with 100% accuracy, including a single example of a metastatic tumour from a primary lung carcinoma. A tumour sample from the collecting duct of the kidney showed a different NMR spectral profile, and pattern recognition indicated that this sample did not classify with the cortical tumours. PMID- 10797635 TI - Metabolism of D- and L-[(13)C]alanine in rat liver detected by (1)H and (13)C NMR spectroscopy in vivo and in vitro. AB - Alanine is the major amino acid utilized by the liver for gluconeogenesis under normal conditions. The metabolism of alanine in rat liver was investigated by means of (1)H and (13)C NMR spectroscopic studies in vivo and in vitro after infusion of L- and D-alanine labelled with (13)C at the carboxyl and methyl group into normal, fasted rats. Valuable information about different metabolic pathways of alanine in rat liver and their regulation under the conditions of gluconeogenesis were obtained. The enrichment of the alanine pool by the infusate was estimated to be 11% for L-alanine and 70% for D-alanine. After infusion of labelled D-alanines, the metabolic pathway via D-amino acid oxidase was observed. The labelled alanines entered the tricarboxylic acid cycle mainly via pyruvate carboxylase; the ratio of pyruvate dehydrogenase activity to that of pyruvate carboxylase is about 28%. The ratio of flux from phosphoenolpyruvate (PEP) through phosphoenolpyruvate kinase as compared with the flux from PEP to glucose was approximately 42%. From the labelling pattern of glucose it was concluded that the pentose phosphate cycle was active under the experimental conditions. PMID- 10797636 TI - Fetal programming of hepatic lobular architecture in the rat demonstrated ex vivo with magnetic resonance imaging. AB - We demonstrate that MRI imaging at sub-millimetre resolution can distinguish between periportal and perivenous zones of the rat liver lobule. This made it possible to measure the hepatic lobular radius in ex-vivo perfused fixed livers using MRI. Comparisons of histomorphometric and MRI measurements of lobular radius were in good agreement, although MRI measurements were significantly smaller (P< 0.001). Male rats whose mothers were fed 40% of the protein of controls during gestation and lactation, had a significantly larger hepatic lobular radius than that of controls [449+/-11 microm vs. 373+/-9 microm (mean +/ SEM), respectively, p<0.001, n = 12; histomorphometry data]. The proton T(2) in periportal and perivenous zones was mapped both before and after antegrade or retrograde perfusion of 10 ml of digitonin (4 mg ml(-1)). Only the T(2) of the hypointense regions increased significantly following antegrade perfusion of digitonin and conversely only that of the intense regions following retrograde perfusion. Digitonin causes permeabilization of cells in specific hepatic zones, determined by the direction of perfusion. The intense and hypointense regions of the hepatic MR images thus arise from the perivenous and periportal zones of the hepatic lobule, respectively. PMID- 10797638 TI - Current Awareness. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of NMR in biomedicine. Each bibliography is divided into 9 sections: 1 Books, Reviews ' Symposia; 2 General; 3 Technology; 4 Brain and Nerves; 5 Neuropathology; 6 Cancer; 7 Cardiac, Vascular and Respiratory Systems; 8 Liver, Kidney and Other Organs; 9 Muscle and Orthopaedic. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. PMID- 10797637 TI - Magnetic resonance spectroscopy of cellular lipid extracts from sensitive, resistant and reverting K562 cells and flow cytometry for investigating the P glycoprotein function in resistance reversion. AB - The proton NMR spectra of K562 cells contain resonances of lipids. When these cells acquire multidrug resistance phenotype, the NMR lipid signals are modified and partially recovered when the resistance is reversed. The goals of the present study are to elucidate the mechanism of the resistance phenotype reversion and to investigate the possible origin of lipid signals detected in whole cells with proton NMR spectroscopy. Therefore, the K562 drug-sensitive cell line, its adriamycin resistant counterpart and two reverting derivates, obtained by verapamil treatment and long term culture in drug-free medium, were used in this study. The P-glycoprotein (P-gp) pump function was measured by flow cytometry and lipids were extracted to be analysed by proton and phosphorus spectroscopy. The phenotype reversion is due to the decrease of the P-gp function and an increased entrance of anthracycline drug when compared with the resistant cells. The spectra obtained on extracts showed no modification of the fatty acid composition and of the ratio of total cholesterol to fatty acid content. A different phospholipid composition in sensitive and resistant cells was found, but the reversion of resistance did not produce a recovery of these lipids. Thus, the lipid NMR spectra of extracts could not explain the spectral modifications observed on whole cells, in relation to acquiring and reverting drug resistance. These results are in favour of a different lipid organization or of localization within the cell. PMID- 10797639 TI - Effects of temperature, incubation period and substrate on production of fusaproliferin by Fusarium subglutinans ITEM 2404. AB - The kinetics of the production of fusaproliferin by Fusarium subglutinans ITEM 2404 in maize and rice cultures was investigated at various incubation temperatures. The growth rate of F. subglutinans was highest at 20 degrees C and 25 degrees C in maize cultures and at 15 degrees C in rice cultures. Although the growth rate was higher in rice than in maize, the maximal production of fusaproliferin was obtained in maize cultures, with a maximum yield (4309 microg g(-1)) at 20 degrees C for 6 weeks. In rice cultures the optimal incubation regimen was at 15 degrees C for 6 weeks, with a fusaproliferin level of 1557 microg g(-1). The production of fusaproliferin at 25 degrees C and 30 degrees C in both substrates was very low, with maximal yield at 25 degrees C of 979 microg g(-1) after 2 weeks and 143 microg g(-1) after 3 weeks in maize and rice cultures, respectively. PMID- 10797640 TI - High performance liquid chromatography for the analysis of fusapyrone and deoxyfusapyrone, two antifungal alpha-pyrones from Fusarium semitectum. AB - A simple, very sensitive and rapid HPLC method was developed for the simultaneous quantitative analysis of both fusapyrone (FP) and deoxyfusapyrone (DFP), the two antifungal 3-substituted-4-hydroxy-6-alkyl-2-pyrones isolated from rice culture of Fusarium semitectum, in crude extracts. Such method was optimized on C-18 reverse phase column, using the isolated metabolites as standards, with a sequence of linear elution steps with a MeOH-H(2)O mixture and using an ultraviolet detector fixed at 285 nm, where both alpha-pyrones showed a characteristic absorption maximum. This method was used to quantify the bioactive metabolites in crude organic extracts from two F. semitectum strains. The recovery of FP and DFP was measured in a crude extract from a poor metabolite producer F. semitectum strain. The recovery values ranged from 84% to 99% for FP and from 99% to 101% for DFP, indicating that the method was close to quantitative recovery. Furthermore, an efficient medium pressure column chromatography and TLC combined method was developed for the isolation and purification of FP and DFP from fungal culture extracts. PMID- 10797641 TI - Comparison between inhibitory indirect ELISA and HPLC methods to quantify free and adducted aflatoxins in human urine. AB - HPLC and Inhibitory Indirect ELISA (I.I. ELISA) methods for quantitation of aflatoxins (AF) in human urine were compared in terms of specificity, sensitivity, easiness and cost. I.I. ELISA was optimized in kind of antibody in use, type of plastic plate, adduct synthesis technique, peroxidase and antibody dilutions, etc. Both polyclonal (Cuban) and monoclonal (British) anti-AF antibodies were statistically studied and the process was standardized. HPLC and electrophoresis were performed while synthetizing AFB(1)-DNA and AFB(1)-Cl Ovalbumin (AFB(1)-Cl-Ov) adducts. Costar polystyrene plate had the best adherence. Optimum coating dilution was 10 ng of AFB(1)-Cl-Ov per well. Dilutions of 1:1000 of monoclonal antibody from purified culture or 1:300 from monoclonal antibody from tissue culture and 1:1000 of peroxidase anti-mouse conjugate were the best. Optimum separation with HPLC was obtained isocratically with 60% MeOH and 40% distilled water mobile phase. ELISA had a sensitivity of 1 pg mL(-1) AFB(1) and HPLC sensitivity was 0.1 ng mL(-1) AFB(1) with fluorescence detector and 4.5 ng mL(-1) with UV detector. Monoclonal antibody gave more accurate results for determination of free and adducted AFB(1) in urine analysis. PMID- 10797642 TI - Confirmation of yessotoxin and 45,46,47-trinoryessotoxin production by Protoceratium reticulatum collected in Japan. AB - Two different strains of the dinoflagellate Protoceratium reticulatum collected at Harima Nada and Yamada Bay in Japan were cultured and analyzed by fluorometric HPLC for yessotoxin production. Only the Yamada Bay strain produced yessotoxin. The toxin together with its analog, 45,46,47-trinoryessotoxin, were isolated from larger scale culture and unambiguously confirmed by (1)H NMR and MS measurements. This is the first confirmation of the biogenetic origin of yessotoxin in Japan, where the toxin was first reported. The results also indicate that the production of yessotoxins by P. reticulatum differs from strain to strain, in a similar way to that observed in many other toxigenic dinoflagellates such as Dinophysis spp. and Alexandrium spp. PMID- 10797643 TI - Neuronal binding of tetanus toxin compared to its ganglioside binding fragment (H(c)). AB - The non-toxin 50 kD C-terminus peptide of the heavy chain of tetanus H(c) contains the ganglioside binding domain of tetanus toxin (TTX). H(c) retains much of the capacity of tetanus toxin for binding internalization and transport by neurons. For this reason tetanus H(c) has been studied as a vector for delivery of therapeutic proteins to neurons. We directly compared H(c) and TTX in the capacity to bind and be internalized by neurons by ELISA. Primary cultures of dissociated fetal cortical neurons were incubated with equimolar amounts of TTX or H(c). Neuronal associated tetanus protein was 4-8 fold greater on a molar basis with tetanus toxin compared to H(c) (1 h incubation). This increase in neuronal tetanus protein was evident with incubation in concentrations from 0.1 microM to 2 microM. There were greater amounts of TTX delivered to the cultured cells at both 0 degrees C (representing membrane bound tetanus protein) and 37 degrees C (bound and internalized tetanus protein). Unlike H(c), TTX showed significant continued accumulation of protein with increasing incubation durations. Neuronal associated TTX increased 2-3 fold over incubation times ranging from 1 to 8 h. Tetanus toxin appears to be clearly superior to the ganglioside binding fragment (H(c)) in the capacity for neuronal binding and internalization. Atoxic tetanus proteins containing additional molecular domains as well as H(c) may be more suitable vectors for linkage with therapeutic proteins and delivery to neurons. PMID- 10797644 TI - Monitoring brevetoxins during a Gymnodinium breve red tide: comparison of sodium channel specific cytotoxicity assay and mouse bioassay for determination of neurotoxic shellfish toxins in shellfish extracts. AB - In October of 1996, a Gymnodinium breve bloom occurred in shellfish harvesting waters of Alabama, Mississippi and Louisiana, Gulf of Mexico, USA. Bloom densities reached 5.6x10(5) cells liter(-1) and bloom residence at shellfish sampling stations ranged from 3 to 28 days. Brevetoxin-2 dominated G. breve toxin profiles in bloom seawater extracts. Shellfish toxicity, assessed by mouse bioassay, exceeded the guidance level for up to 75 days after the bloom had dissipated. Cytotoxicity assays and mouse bioassays showed similar temporal patterns of shellfish toxicity, but the two methods differed in estimations of brevetoxin-3 equivalent toxicity by a factor of 93 to 1. LC-ESI-MS showed the temporal patterns in shellfish toxicity reflected metabolism of G. breve toxins. The molecular ions m/z 1004, 1017 and 1033 dominated LC-ESI-MS spectra of toxic chromatographic fractions from the extracts and were identified as brevetoxin metabolites on the basis of LC-APCI-MS-MS. The discrepancy between cytotoxicity and mouse bioassay estimates of brevetoxin-3 equivalent toxicity resulted from the difference in extraction efficiency of solvents used in the respective methods and the relative sensitivity of the assays to toxin metabolite mixtures present in the extracts. The normalized cytotoxicity assay showed 75% agreement with mouse bioassay positive test samples and 64% agreement with mouse bioassay negative test samples. Published in 1999 by John Wiley & Sons, Ltd. PMID- 10797645 TI - A French monitoring programme for determining ochratoxin A occurrence in pig kidneys. AB - Ochratoxin A is a carcinogen and nephrotoxin which can enter the food chain resulting in human exposure. As pig herds are exposed to ochratoxin A through their feed, their kidneys, livers and pork meat are considered as a possible route of exposure for humans. France, an important producer of pork and pork products, set up a national monitoring programme which included the training of six routine public laboratories in the analysis of ochratoxin A using an immunoaffinity step followed by a HPLC-fluorimetric detection. The programme randomly sampled 300 healthy and 100 nephropathic pig kidneys in 1997 and 710 healthy pig kidneys in 1998. Less than 10% of samples were significantly contaminated by ochratoxin A : in the 1997 survey, 1% of samples contained 0.40 1.40 microg kg(-1) of ochratoxin A and in the 1998 survey 7.6 % exhibited ochratoxin A levels in the range 0.5-5 microg kg(-1). In the case of nephropathic kidneys, only traces of ochratoxin A (0.16 to 0.48 microg kg(-1)) were detected in six samples out of 100. Even if not a major route of exposure for humans, pigs are clearly exposed to this mycotoxin and monitoring of pork products and of feed for swine is necessary. PMID- 10797647 TI - Determination of (R)- and (S)-ketoprofen in human plasma by liquid chromatography/tandem mass spectrometry following automated solid-phase extraction in the 96-well format. AB - A sensitive and selective method was developed for the determination of (R) ketoprofen ((R)-kt) and (S)-ketoprofen ((S)-kt) in human plasma using chiral liquid chromatography/tandem mass spectrometry (LC/MS/MS). Plasma samples spiked with stable-isotope-labeled [(13)C(1), (2)H(3)]-(R and S)-ketoprofen, for use as the internal standards, were prepared for analysis using automated solid-phase extraction (SPE) in the 96-well microtiter format. The enantiomers were separated on an (R)-1-naphthylglycine and 3,5-dinitrobenzoic acid (Chirex 3005) 250x2.0 mm i.d. analytical column, equipped with a 30x2.0 mm i.d. guard column using isocratic mobile phase conditions. The (R)- and (S)-kt levels were quantifiable from 0.05 to 2500 ng ml(-1) by constructing two separate curves from calibration standards covering the same range. The first curve ranged from 0.05 to 100 and the second from 100 to 2500 ng ml(-1). A concentration of 0.05 ng ml(-1) of either enantiomer was easily detected using a 1 ml plasma sample volume. The average method accuracy, evaluated at four levels over an extended period, was better than +/-3% over the entire range. The precision for the same set of quality control samples ranged from 4.0 to 7.0 % RSD (n = 24). The method was applied to the evaluation of pharmacokinetic parameters in human plasma obtained from volunteers who received 25 mg of kt by peroral administration of Actron caplets or by topical administration of Oruvail gel. PMID- 10797648 TI - Mass spectrometric methodology for the analysis of highly oxidized diterpenoid acids in Old Master paintings. AB - Diterpenoid resins from larch and pine trees and the corresponding fractions in a >100-year-old wax-resin adhesive and varnish and a 200-year-old resin/oil paint sample were analysed with by gas chromatography/mass spectrometry (GC/MS) using several off-line and on-line derivatization methods. The main resin compounds were highly oxidized abietic acids. Important products found are hydroxydehydroabietic acids (OH-DHAs), 7-oxoDHA, di-OH-DHAs and 15-OH-7-oxoDHA. The last two compounds have not been reported to occur in artworks before. Larixyl acetate, an important marker from larch resins, was found to be still present in high amounts in the adhesive. A large number of mass spectra of the different oxidation products and larixol and larixyl acetate are presented and their fragmentation behaviour under electron impact conditions is discussed. An index for the degree of oxidation (IDOX) of the abietic acids is presented as an indicator of the degree of oxidation of the matrix in which the resin is present. The IDOX was 0.10, 0.67, 0.81 and 0.76 for the fresh resins, the dark-aged adhesive, the aged varnish and the resin/oil paint, respectively (measured with pyrolysis (Py)-tetramethylammonium hydroxide (TMAH)-GC/MS). Py-TMAH-GC/MS and direct temperature-resolved mass spectrometry are reliable, valuable and fast techniques for the assessment of the presence and degree of oxidation of diterpenoid resins. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10797646 TI - Contributions of mass spectrometry to structural immunology. AB - Mass spectrometry has made important contributions to the field of immunology in the past decade. A variety of mass spectrometric-based techniques have been applied to study the structures of macromolecules that play a vital role in the immune response. These include traditional molecular mass measurements to identify post-translational modifications and structural heterogeneity, mass mapping of proteolysis products, sequencing by tandem mass spectrometry and conformational analysis. Antigen-antibody and other immune complexes have been detected by mass spectrometry, providing an avenue to study macromolecular assemblies that are important to immune function. By virtue of the ability of mass spectrometry based techniques to analyze complex biological mixtures, mass spectrometry has also been employed to identify and sequence protein epitopes important in both the humoral and cellular immune responses. This has been achieved through a combination of immunoaffinity and mass spectrometric techniques, and the coupling of high-performance chromatographs to mass spectrometers. These approaches are important for the identification of pathogens and show promise for the early diagnosis of disease associated with viral and bacterial infection and malignancy. These investigations will enable the mechanisms associated with normal and impaired immune function to be elucidated. Mass spectrometry has been utilized to characterize the structure of peptide mimics, multiple antigenic peptides and other constructs in the design of synthetic immunogens. Information derived from these studies will aid in the development of novel therapeutics and vaccines. PMID- 10797649 TI - Experimental determination of the alpha and beta C--H bond dissociation energies in naphthalene. AB - The acidities of the two different sites in naphthalene (1alpha and 1beta) and the electron affinities of the alpha- and beta-naphthyl radicals were measured using a Fourier transform mass spectrometer. Both carbon-hydrogen bond dissociation energies for naphthalene also were obtained, in this case via the application of a thermodynamic cycle. The final results are DeltaH(o)acid (1alpha) = 394.2+/-1.2 kcal mol(-1), DeltaH(o)acid (1beta) = 395.5+/-1.3 kcal mol(-1), EA(alpha) = 31.6+/-0.5 kcal mol(-1), EA(beta) = 31.6+/-0.5 kcal mol(-1), BDE(1alpha) = 112.2+/-1.3 kcal mol(-1) and BDE(1alpha) = 111.9+/-1.4 kcal mol( 1), and they are compared to benzene and phenyl radical as well as ab initio and density functional theory (B3LYP) calculations. PMID- 10797650 TI - Characterization of prenylated xanthones and flavanones by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. AB - Reversed-phase liquid chromatography with atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) in the positive-ion mode was utilized to analyze crude ether extracts from the root bark of Maclura pomifera, a tree known to have a high content of prenylated xanthones and flavanones. Identification of three xanthones and two flavanones was based on their unique mass spectra. Under optimum conditions peaks corresponding to the [MH](+) ion and characteristic fragments for each compound were observed. (1)H NMR data were used to confirm the identities of two xanthones that had the same molecular mass and similar fragmentation patterns. Fragmentation of the analytes was achieved by application of an electrostatic potential at the entrance of the single quadrupole mass spectrometer. The optimum voltage for fragmentation was found to be related to the class of compounds analyzed and, within each class, to be dependent on the structure of the prenyl moiety. Collision-induced pathways consistent with precedent literature describing the MS characterization of similar compounds and with the observed fragmentation patterns are tentatively proposed. PMID- 10797651 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of some conducting polymers. AB - The application of laser desorption/ionization and matrix-assisted laser desorption/ionization mass spectrometry to the study of the structure of some conducting polymers was investigated. A methodology was developed and the experimental conditions for the characterization of these polymers were established. The parameters chosen to be changed for the method optimization are the matrix material (seven different matrices were tested depending on the polymer studied) and the solvent used in the preparation of the samples. The method seems to be suitable for the study of the structure of two oxidation states of polyaniline: emeraldine and pernigraniline. The analysis of the polymers derived from m-aminophenylbenzothiazole, benzothiazole and tetrathiopentalenes was also performed.} Copyright 2000 John Wiley & Sons, Ltd. PMID- 10797652 TI - Identification of phosphorylation sites in proteins by nanospray quadrupole ion trap mass spectrometry. AB - A method is described for identifying serine phosphorylation sites in proteins, based on conventional (32)P labeling followed by electrophoretic separation, 'in gel' digestion with a protease, peptide extraction, reversed-phase high performance liquid chromatographic separation and collection and off-line analysis of the radioactive fractions by nanospray ion trap mass spectrometry. The method was successfully applied to the identification of three phosphorylation sites in two proteins which were subjected to in vitro phosphorylation under physiological conditions. Different combinations of the various scanning modes of the ion trap, including high-resolution, multiple subfragmentation (or MS(n)) and fast scan analysis, were employed to identify the phosphopeptides, determine their sequence and localize the exact site of phosphorylation. 'Blind' fragmentation using fast scans was used to analyze a phosphopeptide which was undetectable in other scanning modes. The sequence, phosphorylation site and double cysteine modification of the potassium adduct of a peptide containing 35 residues were also determined by multiple fragmentation. The results not only support the validity of the proposed method for routine identification of phosphorylation sites, but also demonstrate the exceptional capability of off-line ion trap mass spectrometry in combination with nanospray ionization for performing very detailed studies on the structure of peptides. PMID- 10797653 TI - Distinction and quantitation of leucine-isoleucine isomers and lysine-glutamine isobars by electrospray ionization tandem mass spectrometry (MS(n), n = 2, 3) of copper(II)-diimine complexes. AB - Electrospray ionization of mixtures of isomeric and isobaric amino acids was investigated with the goal of distinguishing and quantifying the components. Isomeric amino acids leucine and isoleucine were readily distinguished and quantified in 90 : 10 to 10 : 90 binary mixtures using two-stage (MS(2)) and three-stage (MS(3)) tandem mass spectrometric dissociations of ternary Cu(2+)-2, 2'-bipyridyl (bpy) complexes, [Cu(AA - H)bpy](+). The complexes self-assembled in solution upon mixing the components and provided a convenient means of efficient derivatization that increased the efficiency of amino acid ionization by electrospray and shifted the mass of the analytes to a region which was free of solvent interferences. Low-energy dissociations of [Cu(AA - H)bpy](+) complexes in a quadrupole ion trap were achieved at >90% conversions and >80% trapping efficiencies for the MS(2) and MS(3) precursor and fragment ions. Isobaric amino acids glutamine and lysine were also distinguished through MS(2) and MS(3) of their ternary complexes with Cu(2+) and bpy. ESI of [Cu(Gln - H)bpy](+) was enhanced in the presence of [Cu(Lys - H)bpy](+), which resulted in non-linear response at low Lys concentrations. PMID- 10797654 TI - Identification of an artifact in the mass spectrometry of proteins derivatized with iodoacetamide. AB - Derivatization of cysteinyl residues is often used to prevent the formation of disulfide bonds during protein isolation and analysis. The most commonly used reagents are iodoacetic acid and iodoacetamide, which increase the molecular mass of the protein by 58 or 57 Da, respectively, for each derivatized cysteine. A possible side reaction is derivatization of methionine. In our analysis of derivatized human lens alphaA-crystallins, we found an apparent molecular mass 48 Da lower than the mass expected for alphaA-crystallin with the cysteines carboxyamidomethylated. Analysis of a tryptic digest of this protein showed that both cysteines and one methionine had been derivatized. Peaks indicating a molecular mass 48 Da less than expected for the protein with only cysteines derivatized were attributed to fragmentation of the derivatized methionine through collision-induced dissociation in the electrospray ionization source. An awareness of this artifact is important to investigators searching for proteins and their modified forms in complex mixtures. PMID- 10797655 TI - Current Awareness. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of mass spectrometry. Each bibliography is divided into 11 sections: 1 Books, Reviews & Symposia; 2 Instrumental Techniques & Methods; 3 Gas Phase Ion Chemistry; 4 Biology/Biochemistry: Amino Acids, Peptides & Proteins; Carbohydrates; Lipids; Nucleic Acids; 5 Pharmacology/Toxicology; 6 Natural Products; 7 Analysis of Organic Compounds; 8 Analysis of Inorganics/Organometallics; 9 Surface Analysis; 10 Environmental Analysis; 11 Elemental Analysis. Within each section, articles are listed in alphabetical order with respect to author (6 Weeks journals - Search completed at 2nd Feb. 2000) PMID- 10797656 TI - HTLV-I associated infective dermatitis may be an indolent HTLV-I associated lymphoma. AB - When present for a first time blood donation, a 28-year-old Brazilian white female reported a pruritic eczema of the scalp and retroauricular areas since childhood that had been frequently infected. Her mother had been diagnosed as having HTLV-I-associated myelopathy (HAM), and the patient was found to be a human T-lymphotropic virus type-I (HTLV-I) carrier. The patient had been breast fed for 6 months. The patient had a complete examination, and a biopsy was taken from eczema in the retroauricular area. The biopsy indicated chronic lymphohistiocytic dermatitis with no abnormal lymphocytes. Eleven months later, the patient had an infiltration in the skin of the retroauricular area and a new biopsy revealed atypical lymphocytes. Nested polymerase chain reaction (PCR) was positive for HTLV-I and immunohistochemistry of the tissue at this time confirmed adult T-cell leukemia/lymphoma (ATLL). Retrospective immunohistochemistry showed that the first fragment submitted from the biopsy 11 months before was also compatible with the diagnosis of ATLL. This case fulfilled all major criteria for diagnosis of HTLV-I-associated infective dermatitis (HTLV-I-ID). We postulate that the patient had indolent ATLL associated with HTLV-I infective dermatitis since childhood. We recommend that tissue immunohistochemistry analysis be done in any patient with HTLV-associated infective dermatitis. PMID- 10797657 TI - Histoplasmosis presenting as acute respiratory distress syndrome after exposure to bat feces in a home basement. AB - This report describes the clinical and epidemiological findings of an outbreak of histoplasmosis in Brazil involving four persons of the same family, one adult and three children. Contamination by fecal material of bats in the basement of the house was found to be the most likely cause of this outbreak. None of the patients had evidence of immune deficiency. The adult died of acute respiratory distress syndrome. A more severe presentation of the disease in this patient may be partly explained by a heavier exposure to the feces of bats. PMID- 10797658 TI - Helping patients who deny pain. PMID- 10797660 TI - Understanding ventilation and perfusion. PMID- 10797659 TI - Protecting yourself from high-risk i.v. devices. PMID- 10797661 TI - Myths & facts ... about pressure ulcers. PMID- 10797662 TI - Y2K planning: where do you fit in? PMID- 10797664 TI - A quick look at antibiotics. PMID- 10797663 TI - When disaster strikes. PMID- 10797665 TI - Responding to tricyclic antidepressant overdose. PMID- 10797666 TI - Remembering Rhonda.... PMID- 10797667 TI - A quick check of the hematologic and immunologic systems. PMID- 10797668 TI - How to manage vital-organ donors. PMID- 10797669 TI - Action stat. Vancomycin reaction. PMID- 10797670 TI - Preventing postoperative complications. PMID- 10797671 TI - Resolving the delirium dilemma. PMID- 10797672 TI - A question of pain. PMID- 10797673 TI - How to handle grief with wisdom. PMID- 10797674 TI - Neurologic emergency! PMID- 10797675 TI - Ankle-brachial index. Calculating your patient's vascular risks. PMID- 10797676 TI - Reining in a runaway infection. PMID- 10797678 TI - Charting with care. PMID- 10797677 TI - Managing heart failure. Teaching your client how to stay at home. PMID- 10797679 TI - Drug challenge. PMID- 10797680 TI - Understanding leukocyte reduction in blood transfusions. PMID- 10797681 TI - A case of patient dumping? PMID- 10797682 TI - All in good time. PMID- 10797683 TI - Understanding your patient's pain tolerance. PMID- 10797684 TI - Administering an insulin injection. PMID- 10797685 TI - Using a sternal intraosseous device in adults. PMID- 10797686 TI - Comparing peripheral venipuncture sites. PMID- 10797687 TI - Taking a hard look at cirrhosis. PMID- 10797688 TI - Mrs. Reilly's prayer. PMID- 10797689 TI - Patient-teaching aid. "Am I having a stroke"? PMID- 10797690 TI - Helping your patient back on track after knee replacement surgery. PMID- 10797691 TI - ALLOWing for cultural diversity. PMID- 10797692 TI - Action stat. Reye's syndrome. PMID- 10797693 TI - Beyond aspirin: how glycoprotein inhibitors ease acute coronary syndromes. PMID- 10797694 TI - Interpreting the ins and outs of pulmonary function tests. PMID- 10797695 TI - Don't become a casualty in the war of the words. PMID- 10797696 TI - Gauging bladder volume--without a catheter. PMID- 10797698 TI - Understanding inflammatory bowel disease. PMID- 10797697 TI - Orthopedic and vascular emergencies! PMID- 10797699 TI - Unforgettable patient. Meeting Peter. Despite a fatal imperfection, the newborn received his mother's love in full measure. PMID- 10797700 TI - Keeping kids healthy. PMID- 10797701 TI - An arresting situation. PMID- 10797702 TI - Risperidone, edema, and more. PMID- 10797703 TI - Protecting pediatric nurses. PMID- 10797704 TI - A nurse's Christmas. PMID- 10797705 TI - Myths & facts ... about deafness. PMID- 10797706 TI - No room at the inn. PMID- 10797707 TI - Patient hygiene: Part 2. Skin care. Keeping the outside healthy. PMID- 10797709 TI - Managing a midline catheter. PMID- 10797708 TI - The waiting game. PMID- 10797710 TI - OSHA's push toward safety. PMID- 10797711 TI - Helping patients stick to an analgesic regimen. PMID- 10797712 TI - Changing a central venous catheter dressing. PMID- 10797713 TI - Blowing the whistle. PMID- 10797714 TI - Action stat. Testicular torsion. PMID- 10797715 TI - On the road to successful i.v. starts. PMID- 10797716 TI - Unscrambling dangerous myths about heart disease. PMID- 10797717 TI - Helping a family through the organ donation process. PMID- 10797718 TI - Nursing2000 salary survey. PMID- 10797719 TI - Resting easy with PAP therapy. PMID- 10797720 TI - Controlling herpes simplex. PMID- 10797721 TI - Myths & facts ... about domestic violence. PMID- 10797722 TI - Power of prayer. PMID- 10797723 TI - Test yourself with sample NCLEX questions. PMID- 10797724 TI - How to recognize a real nurse. PMID- 10797725 TI - Listen for the "whispering disease". PMID- 10797726 TI - Keeping in touch. PMID- 10797727 TI - Do I have breast cancer? PMID- 10797729 TI - Nutrition matters. PMID- 10797728 TI - Stop cervical cancer in its tracks. PMID- 10797730 TI - [A first in the Austrian Nursing Journal: the editorial staff is testing for you- the alginate test from the user's point of view]. PMID- 10797731 TI - [Care of a patient with a chronic wound]. PMID- 10797732 TI - [Aspects of the treatment of chronic wounds]. PMID- 10797733 TI - [Confusion about the confused]. PMID- 10797734 TI - [All true life is encounter. 1: Nicole--a case model for successful basal stimulation]. PMID- 10797735 TI - [The aims of rehabilitation in nursing. 2: Experiencing body and soul as a unit]. PMID- 10797736 TI - [Dealing with the hard-of-hearing: speak slowly, loudly and clearly]. PMID- 10797737 TI - [Prevention of violence in hospital geriatric care: the dignity of the elderly is fragile--report from a work shop]. PMID- 10797738 TI - [Ethics in the hospital: a new trend in health care?]. PMID- 10797739 TI - [Quality of health facilities: does control by health insurance inspectors uncover deficiencies?]. PMID- 10797740 TI - [The Internet in the hospital: server, browser, website]. PMID- 10797741 TI - [Rules for visitors in intensive care units: a fossil in a high-tech environment?]. PMID- 10797742 TI - [Malpractice: not all physicians' activities may be delegated]. PMID- 10797743 TI - [How do patients experience the behavior of the nursing personnel after disclosure of the diagnosis of a chronic life threatening disease?]. PMID- 10797744 TI - [Necessary criticism has to be followed by effective measures]. PMID- 10797745 TI - [In favor of ecology and economy. Future recycling concepts]. PMID- 10797746 TI - [Patient orientation and efficiency in nursing. Foundation prize for exemplary projects]. PMID- 10797747 TI - [Cost savings in old age homes: fight about opinions on effective management]. PMID- 10797748 TI - [Additional qualification in professional pedagogics: practical mentoring is no minor matter]. PMID- 10797749 TI - [Thinking--feeling--caring: experiences with intuitive, practical and theoretical knowledge of nursing]. PMID- 10797751 TI - [Illness, sexuality and care: tips for dealing with a tabu]. PMID- 10797750 TI - [Alternative skin care: fit and revitalized with the aid of dry brushing]. PMID- 10797752 TI - [Nursing visit with patients with mental disorders--3. Introduction and transformation are causing new (solvable) problems]. PMID- 10797753 TI - [Continuing education as a specialty counsellor: future chances in geriatrics]. PMID- 10797754 TI - [Promotion prize "Care of the Elderly" 1999: students "travel" with the elderly into their childhood]. PMID- 10797755 TI - [Instructions for administration can cause problems: news about sales tax exemption for mobile nursing services]. PMID- 10797756 TI - [Effect of color therapy on health and wellbeing: colors are more than just physics]. PMID- 10797757 TI - [Mind mapping as a creative method for nursing management: mind maps help to save time]. PMID- 10797758 TI - [The importance of nursing documentation: wrong placement is lacking documentation]. PMID- 10797759 TI - [The principle of continuous improvements: potential of converting of KAIZEN to the service part in the hospital]. PMID- 10797760 TI - [Promotion of professional rehabilitation: retraining to be a nurse is refused for health reasons] [In Process Citation] PMID- 10797761 TI - [Quality assurance by means of care planning--a worthwhile proof?]. PMID- 10797762 TI - [The second stage of the Nursing Insurance Act and its effects on the institutional care within welfare for the elderly]. AB - OBJECTIVE: The intention of the recently introduced compulsory nursing care insurance in Germany has been to support patients as well as relatives, and to reduce the burden on the social insurance system in financing professional services. In this study we examine the consequences of the 2nd stage of this legislation upon the care in old people's homes. METHODS: The investigation covered 5 institutions comprising two components: 1) Analysis of the records kept by the administration (N = 369), and 2) analysis of the volume of nursing care in a sample of N = 121 (including basic care as well as psychosocial support) provided by the staff over a period of 14 days. RESULTS: The required volume of basic nursing care (SGB XI) determines the classification of patients according to their need for nursing. Although by law the amount of nursing in stage 0 should be none, nevertheless 20 minutes of basic care in average are provided each day here. In stages II and III the observed nursing-time exceeded the allowed values slightly. In stage III realized nursing was far below the legal provision, although the private co-payments were highest. CONCLUSION: The agreement between nursing providers and nursing insurance aiming at comprehensive care, under the given circumstances cannot be executed satisfactorily. This is true especially for stage III patients in old people's homes. PMID- 10797763 TI - [The effect of diagnosis-related groups on hospital care]. AB - OBJECTIVE: The implementation of Diagnosis Related Groups (DRG's) is intended to lead to a shortening of hospital days. DRG's therefore may endanger the quality of hospital care or cause a transfer of services from inward to ambulatory care. In this study the consequences of two selected DRG's are examined: operations of patients with varicosis and with hernia. METHODS: A retrospective comparison of 875 patient files in 6 hospitals in Northrhine-Westphalia (NRW) and a follow-up questionnaire to the patients themselves (N = 510). RESULTS: Between 1995 and 1997 average hospital days have been reduced by 2 days (varicosis) and 1 day (hernia) respectively. However, the standard stay according to an expert commission of the MOH is still exceeded. The shortening of inward care leads to an increase of ambulatory care. Also the patients indicate a worse subjective health status at discharge although this does not carry through the ambulatory phase. CONCLUSION: For the two DRG's examined the reduction of hospital services due to shorter inward periods is mostly compensated during ambulatory care. An additional shortening of hospital days, however, is likely to lead to negative effects on the health status of patients. PMID- 10797764 TI - [Care management: scientific thoughts on a current topic]. AB - The following article deals with one of the management concepts now undergoing extensive international debate--the care management concept favored in Great Britain--and discusses its importance in respect to the optimization of outpatient nursing care in the Federal Republic of Germany. After a presentation of the key elements comprising the concept, four central task complexes relating to care management are discussed: establishing demand--and need oriented nursing, assuring the integration and continuity of care, especially when chronic diseases are involved, helping family members to provide nursing care and promoting health as one of the nursing profession's tasks. It is shown that care management is a tool that can be used to tackle deficits in outpatient nursing care and find innovative solutions to them, thereby promoting the nascent professionalization of nursing in Germany, a trend that is long overdue in international terms. PMID- 10797765 TI - [Health- and care reporting in Germany. State and perspectives]. AB - The first Health Report for Germany gives a good opportunity to look for further perspectives of the development of health and care reporting. This article refers briefly to concept, historical development, the report itself and critical analyses of contents, political function and organisational structures of health reporting in Germany. These have a similar importance for care monitoring. The article proposes a stronger participation of care reporting in health reporting not only in Germany and a stronger differentiation between nursing research and care reporting. PMID- 10797766 TI - [Evaluation of courses in nursing science in Germany. A pilot project on the current state and on perspectives of future development]. AB - Since 1991 more than 40 courses of nursing science have been established in Germany. They differ mainly in two aspects: a) they have different course contents and b) they belong to different faculties. This study focuses on two questions. (i) How is the integration of scientific knowledge proceeding? (ii) Which positive aspects and problems can be identified? For this purpose we used a questionnaire to ask heads of the faculties from all courses of nursing sciences. 32 questionnaires were returned. We have come to the conclusion that there is a broad scientific basis for courses of nursing science in Germany. PMID- 10797768 TI - [Evidence-based nursing: a method for nursing practice?]. AB - Evidence-based nursing is the concept of critically appraising the best evidence from research. It integrates research evidence with clinical expertise into decision making in nursing care for patients. The method is derived from the new paradigm in medicine--evidence-based medicine, developed as a learning method by epidemiologists at the McMaster University in Canada. This article states the necessity of this method for nursing. With the help of a clinical scenario (fall prevention in the elderly) the author explains how to use evidence-based nursing in practice. PMID- 10797767 TI - [Knowledge of professional rules of ethics among professional nurses]. AB - During recent years numerous publications on ethical issues appeared in German nursing literature. Nursing theorists repeatedly refer to the Code of Ethics of the International Council of Nursing (ICN). The implementation of codes of ethics is discussed as an indispensable stage within the development of independent nursing ethics. However, it is not clear whether nurses in Germany know about the ICN Code of Ethics or other codes of ethics. The authors' objective is to learn more about how much is known about codes of ethics by practical nurses in four selected hospitals. In addition, it is of interest to know, which advantages nurses might associate with codes of ethics and secondly from which sources nurses prefer to receive ethical education. The study shows that only 25% of the participants know about codes of ethics. It also reveals that further education offered by hospitals or the basic nursing education are seen as the most important source of information related to ethics. The major advantage of codes of ethics seen by the participants is to provide guidance in the decision-making process of ethical dilemma situations in nursing. Secondly the Code of Ethics is regarded as a useful framework to define obligations and rights of nurses in the relationship with patients and relatives. The results of the study indicate that the ICN Code of Ethics is practically unknown among nurses and should be discussed more extensively by practical nurses and nursing theorists. PMID- 10797769 TI - [Putting it into practice is difficult. A qualitative study on the theory practice transfer in the case of kinesthetics]. AB - The theory-practice-gap in nursing is a well-known phenomenon. However, to date there is only very little well-founded evidence in nursing research available on this topic, especially in the context of the FRG. Therefore, a qualitative nursing research project, oriented by the methods of Grounded Theory, on theory transfer into practice using the example of a continuing in-service education program on kinesthetics in nursing was carried out that aimed at examining factors that influence the implementation of kinesthetics into nursing practice. After the program, the researcher conducted mainly semi-structured interviews with the participants about their experiences with using what they had learned in their daily work on the ward. As part of the results, the impact of the category "attributes of kinesthetics" on the theory-practice-transfer of the participants in the continuing education program on the topic will be shown. That is to say something about whether the fact that kinesthetics is perceived as practical, strange and exotic, connected with closeness, or as difficult or risky, has an influence on the use of newly learned knowledge in practice. Finally, the relation of this category to the key-category "working in knowledge and new things" will be shown. The findings of this study suggest that putting emphasis on transfer of knowledge alone is not sufficient to account for the complex structure of the process of theory-practice-transfer. PMID- 10797770 TI - [Patients with diaphragmatic pacemakers]. AB - Due to the implantation of diaphragmatic pacemakers, the hope and quality of life for those with upper spinal column injuries has increased considerably. Until about 10 years ago, these patients depended on a mechanical ventilator with all of its corresponding limitations. Nurses have had to learn how to use diaphragmatic pacemakers so they may provide patients with the specific treatment these make available and so that they may offer adequate health education to relatives of these patients. The greatest challenge for our profession is to succeed in having these patients abandon the hospital and integrate into society again. This article analyzes how diaphragmatic pacemakers work and the specific nursing treatment these patients require, placing emphasis on psycho-sociological aspects. PMID- 10797771 TI - [Generic medicines]. AB - The cost increases for pharmaceuticals obliges our political and health care officials to attempt to contain these costs. One possible measure which must be carefully taken under consideration is to apply a policy using generic medicines which ensures the quality, efficiency and security aspects of generic medicines. In this article, the author reviews the concept and definition of what is the generic pharmaceutical specialty; the requirements necessary to comply with in order for a medication to be classified as generic; the current status of these medications in Spain; the factors regarding their prescription, dispensation and substitution for established medications; and the advantages and disadvantages of their use. PMID- 10797772 TI - [Female rituals: preceding maternity]. AB - Throughout our lives, we pass through important stages of change, from the moment of our births until the moment of our deaths. These experiences entail social rituals. Pregnancy and childbirth are one of these changes in which women will encounter multiple rites, defined mainly by medical practices and beliefs. Every society has its rites which work to maintain an ideology. With a rite, we recognize all that a society considers correct. A woman during pregnancy will undergo those actions and interventions considered normal in this ritual process, with the final objective being to achieve motherhood. PMID- 10797773 TI - [The hospital nursing discharge form. Establishing and validation of a standard formate]. AB - The purpose of this article is to describe the design, application and validation process for a new hospital nursing medical discharge form. This process has three phases: FIRST PHASE: Redesigning the Document. In 1996, as a response to growing demands as well as the need to adapt to the requirements of data processing, all nursing documents in all hospital nursing files were updated following the recommendations of the Technical Commission. SECOND PHASE: Initial Application and Validation. The medical discharge form prepared in phase one was applied during the first trimester of 1997. The parties involved agreed to test this form until the end of the year in order to come to a consensus regarding its structure and content while at the same time determining its degree of comprehension and usefulness. THIRD PHASE: Editing and Final Format. The results of these analyses, together with revisions of criteria included in the NMDS and from the Conference on Hospital Discharge Abstract System, made it possible to draw up the final draft of this form which include nursing discharge criteria in the evaluation section. A pilot test was carried out in five hospital units to determine the validity of this form according to the same criteria of comprehension and usefulness. The results indicate a recommendation to eliminate the sections of medical diagnosis and medication upon discharge while to maintain the identification of the patient, the summary of his/her stay in the center, the description of the patient's case upon discharge and the treatment plan. PMID- 10797774 TI - [Children with AIDS. Treatment plan and procedure of care]. AB - The authors discuss treatment provided to children suffering from Acquired Immunodeficiency Syndrome (AIDS) and to their families. By means of checking the nursing files for hospitalized children, the authors observe alterations and omissions which are common to other diseases, plus those which are specifically related to this disease: psychosocial problems. A treatment plan and an assistance procedure are drawn up. In these, the role the family plays to support the patient as well as that of the multidisciplinary team are emphasized. PMID- 10797775 TI - [Subcutaneous drug administration. Indications for the use in terminal patients]. AB - The preferred methods to administer medicines to the sick are oral, under the tongue, or rectal; but in the case of the terminally ill patient, we may encounter the problem that the drugs which they tend to require do not come in either of these preferred administering methods since these patients symptoms and problems demand complex responses. In these cases, the subcutaneous way winds up being highly recommended; therefore, the authors present the pharmacological bases of this method, they describe continuous subcutaneous perfusion, and all those aspects of this method which might be interesting to the professionals who employ it. PMID- 10797776 TI - [Professional development: the mission (II)]. AB - In the second part of this generic article whose purpose is to analyze what is a mission statement of an organization, the author concentrates on two fundamental points: what are the bases upon which a mission statement is constructed from a strategic perspective and what are the functions and advantages of a mission statement. With respect to this first basis, the author points out that an organization's function should be based on sound business criteria; that a mission statement should harmonize with the organizational milieu, reflect the values of the organization's professional staff while paying attention to special interest groups; and focus on professional know-how and on what works well and makes sense for the client. Concerning this second aspect, this article deals with maintaining the integrity of an organization, motivating professionals or utilizing a mission statement as a means of control. PMID- 10797778 TI - [Abdominal viscera (spleen, adrenal glands and large vessels)]. PMID- 10797777 TI - [Bipolar disorder. Follow-up of lithium therapy in a clinic]. AB - Lithium carbonate is the drug of choice in the prophylactic treatment of bipolar disorder. The purpose of this article is to describe the effective functioning of lithium in a clinical way in our hospital under the control of university graduates in nursing. During the first ten months of its use, 40 patients have been included which led to 122 appointments, absenteeism being 8.62%. 20% (8/40) of the lithium intoxications at the start of the follow-up were inferior to 0.5% mmol/l while at the end, only 5.3% (2/38) fell below 0.5% mmol/l. The average of the lithium intoxications at the end of the study period was significantly higher than in the moment of entrance to the program (p = 0.009). During the study period, three relapses were registered. PMID- 10797779 TI - [Research in theory and practice]. AB - By means of a survey of international articles, the authors propose to demonstrate some factors which bear an influence on the development of research and investigation in nursing, some problems which affect it, and some strategies for its development inside a clinical-teaching environment. DEVELOPMENT OR RESEARCH AND INVESTIGATION IN NURSING: In the United States, Canada, United Kingdom, other European countries, especially Spain, the authors observed the strong relationship which exists between the development of research and investigation in Nursing and its incorporation in universities. RESEARCH AND INVESTIGATION IN CLINICS: In the opinion of many authors, the main field for research and investigation lies in clinical practice. The authors analyze seven basic requirements: academic curiosity; education in research and investigation; investigative activity; climate conducive to research and investigation; funds for research and investigation; dissemination of the results of research and investigation activities; and making use of the discoveries. TEACH AND INVESTIGATE: The authors point out strategies which enable a teacher to teach from an investigative approach and to stimulate students towards investigation at various levels according to the circumstances involved. PMID- 10797780 TI - [Nursing care in osteoporosis. Health education and primary care]. PMID- 10797781 TI - [Experimental study. Reduction of pressure in areas of risk of developing pressure ulcers with a hydrocellular dressing]. AB - INTRODUCTION: The handling of pressure is a basic measure in the prevention and treatment of bed sores. It is possible to reduce and ease pressure by various means including changes in posture, use of special surfaces for handling pressure as well as the use of local applications or external applications which reduce pressure. Today nurses have a large quantity of external applications available to use although only some hydrocellular ones are capable to reduce pressure due to their hydrocellular structure. METHODS AND MATERIALS: An experimental study was designed to calculate the level of pressure before and after applying an Allevyn hydrocellular external application in the area of the sacrum, ischium, and heel of three healthy volunteers; first, Volunteer A, a 85 kg. 170 cm man; second, Volunteer B, a 54.3 kg. 159 cm woman; and third, Volunteer C, a 69.4 kg 164 cm man. Measures were taken on two types of surfaces: a viscoelastic foam mattress and a conventional hospital mattress. All measurements were repeated at 0, 30, 45 and 60 degrees of inclination. Pressure was determined by means of a Talley pressure monitor, Oxford Pressure Monitor MK II. RESULTS: A total of 144 pressure reading were taken. The overall average reduction after applying a external hydrocellular application on all volunteers, at all inclinations and on all surfaces for each of the three zones were 19.5% in the sacrum, 13.8% in the ischium and 20.15% in the heel. COMMENTARY: Even though our study has its limitations, such as young, healthy volunteers, we can establish that the external hydrocellular application studied does have a local reducing effect on pressure. Since every external hydrocellular application has its own specific structure, the results of our study can not be applied with certainty to other external applications inside the hydrocellular group. PMID- 10797782 TI - [Recording cardiorespiratory arrest by the Utstein method]. AB - The author introduces a "Case History Form to Serve to Follow the Process of Cardiorespiratory Arrest" drawn up by means of the recommended guidelines for uniform data communication in cases of cardiorespiratory arrest outside the hospital, following the Utstein method. At the same time, given that there still are no set guidelines in the hospital milieu, the author includes a form on which to record the subsequent actions carried out during cardiorespiratory arrest. This case history form was presented orally at a Congress on Resuscitation in Seville. PMID- 10797783 TI - [Functional autonomy and use of leisure time by the elderly]. AB - The authors evaluate the relationship among the degree of functional autonomy and use of leisure time or activities, including an analysis regarding the influence of a patient's sex. To carry out this evaluation, the study used the "EVA" scale, or the Functional Autonomy Scale of Values, on 43 elderly patients over 65 years of age who receive at least one home visit per month. Among the conclusions of this study, the authors deduce that the degree of functional incapacity bears an influence on the use of spare time without observing any significant relationship as to the patient's sex. The authors make some recommendations as to how to improve the EVA scale. PMID- 10797784 TI - [The conflict of tomatoes and olive oil within a bureaucratic machinery]. AB - In this short article, the author relates, by means of a case report, a type of conflict which occurs as a result of the paradoxes derived from the current managerial model practiced in the Andalucian public hospitals. The limitations and the incompetence of a bureaucracy, rigid in compliance with its functions, and which is incapable of attending to the demands that belong to the hard core of the organization, place the supposed most important and most immediate objectives of an efficient managerial system in "danger". The basic hypothesis lies in the need to bear in mind, at the time of designing a hospital's organizational system, the "structures of the invisible interests", with their corresponding paradoxes and perversions, of each of the distinct component parts of this system. And how these invisible bureaucratic interests can hinder and pervert managerial and organizational designs which appear to be ideal and optimal in their objectives and principles. PMID- 10797785 TI - [Operating room nurses: pioneers in quality work]. PMID- 10797786 TI - [Competence. A unique responsibility ... but jointly]. PMID- 10797787 TI - [The necessity of preoperative care]. PMID- 10797788 TI - [The group of operating room nurses at the Percy Military Hospital]. PMID- 10797789 TI - [Equipment: thanks to electronics]. PMID- 10797790 TI - [HIV: the Defraissy report, needle stick injuries, the nurse's reality, the anti AIDS vaccine...]. PMID- 10797791 TI - [Protocol: how to avoid accidents of blood exposure]. PMID- 10797792 TI - [Facing AIDS: experience of a nurse. Interview by Jean-Marie Manus]. PMID- 10797793 TI - [Betting on the International AIDS Vaccine Initiative: an anti-AIDS vaccine in 10 years]. PMID- 10797794 TI - [Occupational nurse: a profession within the profession]. PMID- 10797795 TI - [Occupational nurse in banking, or how to help victims of aggression]. PMID- 10797796 TI - [Vaccines and vaccination: a short history of vaccination]. PMID- 10797797 TI - [Vaccination and the nurse's role]. PMID- 10797798 TI - [Mechanism of action of vaccines and their classification]. PMID- 10797799 TI - [Methods of vaccine administration]. PMID- 10797800 TI - [The new vaccination schedule and the various vaccinations]. PMID- 10797801 TI - [Recommendations for specific vaccinations]. PMID- 10797802 TI - [Obligatory vaccinations for health professionals]. PMID- 10797803 TI - [Vaccines and voyages]. PMID- 10797804 TI - [Adverse effects and contraindications of vaccines]. PMID- 10797805 TI - [Depressive disorders in children]. PMID- 10797806 TI - [Education of nursing personnel on transfusion safety]. PMID- 10797807 TI - [The future of xenotransplantation]. PMID- 10797808 TI - [Smoking cessation, yes, but how to remain a non-smoker?]. PMID- 10797809 TI - [Secrets in home hospitalization. Interview by Marc Giraud]. PMID- 10797810 TI - [A sick nurse is telling you about chronic pain]. PMID- 10797811 TI - [A fundamental biologic rhythm]. PMID- 10797812 TI - [Classification of sleep disorders in children and adolescents]. PMID- 10797813 TI - [Consultation on sleep patterns in infants]. PMID- 10797814 TI - [Respiratory risk factors in children during sleep]. PMID- 10797815 TI - [Polysomnographic examination during rest]. PMID- 10797816 TI - [The quality of sleep in hospitalized children]. PMID- 10797818 TI - [A policy for the care of violent adolescents]. PMID- 10797817 TI - [Therapeutic education: playing "little train" for falling asleep]. PMID- 10797819 TI - [Minor surgeries in children. Mutilate in order to invigorate]. PMID- 10797820 TI - [Refractory shock after converting enzyme inhibitor administration. Usefulness of angiotensin II]. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors are commonly used during the early phase after myocardial infarction but severe hypotension and shock have been described. CASE REPORT: A 42-year old woman underwent a conservative management for an anterior acute myocardial infarction, initially associated with a pulmonary edema. Two hours after the initiation of a treatment with ACE inhibitor administered orally (lisinopril, Zestril), a circulatory failure in conjunction with an acute renal insufficiency occurred. Right heart catheterization disclosed markedly decreased systemic vascular resistance in the presence of a preserved cardiac index. Repeated fluid challenges and intravenous administration of norepinephrine failed to improve the hemodynamic status. The refractoriness of shock raised the hypothesis of a dysregulation of the renin angiotensin system, secondary to the treatment by ACE inhibitor. Accordingly, the patient was given angiotensin II intravenously (Hypertension) which markedly raised systemic vascular resistance, and result in subsequent regression of shock. The patient was discharged after an otherwise uneventful course. DISCUSSION: We reported a refractory shock to fluid challenges and norepinephrine after the first dose of ACE inhibitor during acute myocardial infarction. Regression of shock was possible only with angiotensin II. PMID- 10797821 TI - [Acute renal failure after amoxicillin crystallization]. AB - BACKGROUND: Drug-induced nephrolithiasis is a rare finding, especially with beta lactamins. We report a case of acute renal failure due to amoxicillin crystallization. CASE REPORT: A 48 year-old woman was admitted because of pneumococcal meningitis. After 4 days on high-dose amoxicillin (320 mg/kg/day), she developed acute oliguric renal failure and amoxicillin crystallization was documented by infrared spectrometry. The outcome was favorable after amoxicillin dosage tapering, together with one single hemodialysis session and further hydratation. DISCUSSION: Amoxicillin is mainly excreted in the urine in its unchanged form. The risk of crystalluria is increased by low urinary pH, low urine output and high-dose of the drug. Such a crystalluria should be accurately identified by infrared spectrometry. PMID- 10797823 TI - [Acute hepatitis during cibenzoline therapy]. PMID- 10797822 TI - [Muscle hematoma in the very elderly receiving low-molecular-weight heparins]. PMID- 10797824 TI - [Antibiotic prophylaxis for esophageal varices]. PMID- 10797825 TI - [Contribution of platelet aggregation inhibitors in the prevention of complications of atherothrombosis]. AB - RECOGNIZED EFFICACY: Platelet antiaggregats have been used with success for more than twenty years in industrialized countries, participating in reducing the number of complications of atherothrombosis. New compounds have appeared on the market over the last 3 years. CLOPIDOGREL: Clopidogrel is a thienopyridine similar to ticlopidine but with less toxicity, particularly hematological toxicity. Compared with aspirin for secondary prevention of atherothrombotic events in high risk patients, clopidogrel provides a significant 8.7% (p < 0.043) reduction in the relative risk of severe complications: myocardial infarction, ischemic stroke, death due to vascular cause. The most remarkable benefit is obtained in patients with atheromatous arteriopathy of the lower limbs: 23.8% reduction in relative risk. STENT AFTER ANGIOPLASTY: A combination regimen using aspirin and ticlopidine the month following stent implantation has allowed a very significant reduction in acute (within 24 hours) and subacute (within 30 days) coronary occlusions. Currently the aspirin-clopidogrel combination tends to be used instead of the aspirin-ticlopidine combination due to the lower hematological risk for a similar effect. GPIIBIIIA INHIBITORS: Injectable GPIIbIIIa inhibitors have changed management of the acute phase of coronary artery disease. Among these compounds, abciximab has the most extensively proven efficacy. It is mainly indicated for severe forms of coronary angioplasty with or without stent insertion. It provides a significant short-term and long-term reduction in severe events (myocardial infarction + death + revascularization). Oral formulations of GPIIbIIIa can be used to maintain and/or amplify the benefits obtained with injectable formulations. They are not currently available on the market. ASPIRIN: Due to its very low cost and low side-effect profile, aspirin plays an essential role in most indications where it remains the most widely used first-line platelet anti aggregate. PMID- 10797826 TI - [Monoclonal antibody immunotherapy for malignant non-Hodgkin's lymphoma]. AB - THERAPEUTIC OPTIONS: There is a need in the management of non-Hodgkin's lymphoma (NHL) for tumor-targeted therapy. Immunotherapy with monoclonal antibodies can be used to reach this goal. These treatments have however often produced disappointing efficacy and so have not been broadly applicable to patients. MODIFIED MONOCLONAL ANTIBODIES: Technological advances have been used to alter mouse monoclonal antibodies genetically, leading to the development of mouse/human chimeric or humanized antibodies with demonstrated advantages of reduced immunogenicity and enhanced ability to interact with human effector cells. Initial phase I/II trials of monoclonal antibodies for relapse or refractory NHL, especially with chimeric anti-CD20 rituximab, demonstrated encouraging results. FURTHER ASSESSMENT: Randomized trials are now necessary to compare these agents with standard regimens and to determine their optimal use. PMID- 10797827 TI - [Biosphonates in oncology]. AB - MECHANISM OF ACTION: Tumor-induced osteolysis or lytic bone disease is mediated by osteoclast activation. Bisphosphonates inhibit bone resorption by reducing osteoclastic activity. INDICATIONS: Bisphosphonates were shown to be effective in treating cancer-related hypercalcemia. Recent large randomized clinical trials have shown the efficacy of bisphosphonates in reducing bone pain, pathological fractures and spinal cord compression for patients with multiple myeloma and breast cancer metastatic to bone. The potential survival benefit from pamidronate in patients with advanced myeloma warrants further study. FUTURE: Future clinical trials will use more potent bisphosphonates (zoledronate, ibandronate) with the ultimate goal of trying to prevent bone metastases. PMID- 10797828 TI - [Chronic intracerebral stimulation and abnormal movements: why and when? Major therapeutic advances in the 90s]. AB - TECHNICAL ASPECTS: Surgical procedures for abnormal movements progressively evolved toward microinvasive techniques performed under stereotaxic control. The next advance was the development of well-tolerated intracerebral stimulation with reversible effects. The quality of the results depends on optimal definition of the stimulation target. Optimally, the surgical protocol includes stereotaxic ventriculography to identify targets followed by electrical activity recordings of the structures crossed by the electrode. Finally, a minute study of the stimulation effects is performed under sterotaxic control while the patient is awake. PARKINSON'S DISEASES: Indications for thalamic stimulation in Parkinson's disease have given way to subthalamic stimulation which improves tremor, bradykinesia and rigidity simultaneously. This result confirms the implication of the subthalamic nucleus in the pathogenesis of Parkinson syndromes. The palladium may be stimulated in Parkinson's patients with major iatrogenic involuntary movements. ESSENTIAL TREMOR: Thalamic stimulation is indicated for essential tremor. The efficacy of chronic stimulation is currently under evaluation in other indications. PMID- 10797829 TI - [Homocysteinemia: role in vascular disease]. AB - HEREDITARY DISEASE: Hereditary anomalies of homocysteine metabolism are quite uncommon and manifest by very high homocysteine levels (> 100 mumol/l) and associated homocysteinuria. The risk of premature cardiovascular disease is high. Clinical, biological and epidemiological data accumulated since the 70 s have demonstrated that a moderately elevated serum homocysteine level favors the development of atherothrombosis. PROVEN RISK: The risk of coronary or cerebral events is 1.5 to 3-fold higher for fasting homocysteine levels above 15 mumol/l. These data show that moderately elevated homocysteine level is a powerful cardiovascular risk factor. Further information is however needed to ascertain its frequency in the population and determine whether it is a truly independent risk factor. THERAPEUTIC OPTIONS: Most cases of moderately elevated homocysteine can probably be explained by gene-environment interactions. Homocysteine levels can be lowered by oral administration of vitamin cofactors implicated in homocystein metabolisms: folic acid, vitamin B6, vitamin B12. PMID- 10797830 TI - [Chronic cardiac insufficiency. Treatment with angiotensin I converting enzyme inhibitors and angiotensin II receptor antagonists]. AB - RENIN-ANGIOTENSIN SYSTEM: This system plays a major role in regulation of blood pressure. Angiotensin converting enzyme inhibitors (CEI) modify the balance between the vasocilator and diuretic properties of bradykinin and the vasoconstrictor and antidiuretic properties of angiotensin II, favoring vasodilatation and natriuresis. Angiotensin II receptor antagonists (ARAII) block AT1 receptors and stimulate AT2 receptors with favorable vasodilator and natriuretic affects. CEI: Converting enzyme inhibitors play an indispensable role in the treatment of heart failure and should be prescribed at high dosage. They have a long-term beneficial effect. ARAII: These compound could play a role in the future, but studies conducted to date comparing ARAII and CEI have been unable to demonstrate superior or equivalent effects with ARAII, and do not warrant their prescription for hypertension. PMID- 10797831 TI - [Treatment of arterial hypertension in the diabetic]. AB - BLOOD PRESSURE CONTROL: More than half of all diabetic patients have high blood pressure. Even more so than in the general population, hypertension compromises the cardiovascular and renal prognosis. Optimal blood pressure control can limit the progression of microangiopathy and macroangiopathy as clearly demonstrated in the HOT and UKPDS studies. For the WHO, the goal is to control pressures < 130/85 mmHg. In this respect, there has been no demonstration of a J-curve relationship between pressure lowering with antihypertension drugs and incidence of cardiovascular events among patients with coronary artery disease included in the HOT study. MULTIPLE DRUG THERAPY: Regular long-term monitoring and, in most cases, multiple-drug regimens, are prerequisites for maintaining pressure figures below 130/85. When elaborating a blood pressure control protocol, it is important to consider the presence of coronary artery disease, suggesting use of beta blockers, or renal disease, which should lead to the use of angiotensin converting enzyme inhibitors. Diuretics play an important role in combination regimens and are indispensable in three-drug protocols or in case of altered renal function. SYSTOLIC HYPERTENSION: Subgroup analyses in the SHEP and SYST-EUR studies demonstrated the importance of treating pure systolic hypertension in diabetics. The protection obtained has the same or even more impact than in the general population. RISK FACTORS: Diabetes control must of course be maintained and coherent management requires taking into consideration all the risk factors, especially smoking and dylipidemia. PMID- 10797832 TI - [Current recommendations on arterial hypertension and its therapy. A critical analysis]. AB - OBJECTIVE: The purpose of this editorial is to give a critical view of recommendations recently published on hypertension. ANALYSIS: Present changes in therapeutic aspects involving both pharmacological and non pharmacological treatment are analyzed together with the development of new compounds. CONCLUSION: It is suggested that the organization of care, more than the discovery of new compounds is the critical point for improvement of cardiovascular risk in hypertension. PMID- 10797833 TI - [Current trends in asthma management]. AB - INHALED CORTICOSTEROIDS: There has been substantial improvement in asthma management over the last few years. Inhaled glucocorticoids (ICS) are the most effective therapy in the treatment of asthma. COMPLEMENTARY TREATMENTS: Additional treatments such as theophyllines, long-acting beta 2 agonists or more recently leukotriene antagonists allow in most cases the maintenance of ICS at safe dosages without substantial adverse effects. COMPLIANCE: Compliance is a major issue that can only be improved using global management strategies in which asthma education plays a prominent role. PMID- 10797834 TI - [What are the safety measures for use of low-molecular-weight heparins in the treatment of deep venus thrombosis?]. PMID- 10797835 TI - [Zinc: concepts on an essential micronutrient]. AB - It is now well known that zinc is an essential micronutrient. Even though much information is available, there are many points that remain to be studied. zinc is absorbed by the intestine and transported in the plasma by albumin, forming a small exchangeable pool, which is rapidly exhausted even in cases of mild deficiency. zinc is essential for the activity of about 70 enzymes among other functions. Therefore, zinc deficiency produces a great number of clinical disorders, the symptoms of which may range between mild up to serious dysfunctions. These problems are corrected by a dietary supplementation with zinc. The lack of a reliable method to determine the zinc nutritional status reflects the problem to estimate the metal recommended dietary allowances, which are difficulty met specially by children, old persons and pregnant women which constitute the main risk groups. Although our knowledge concerning zinc toxicity is scarce, it is well known that the amounts of zinc that produces toxic effects are much higher than those that are contained in regular diets as well as in diets supplemented with this metal. Therefore, the need of the development of effective strategies, like food fortification with proper zinc compounds, appears as an attractive alternative in order to prevent and/or correct the deficiency of this vital element and to improve the health and the life quality of the whole population. PMID- 10797836 TI - The role of xanthine oxidase and the effects of antioxidants in ischemia reperfusion cell injury. AB - During last years considerable interest has been devoted to understand the role of oxygen radicals in the ischemia induced cell injury associated with reperfusion. In the brain and in others tissues, free radicals play a role as modulators of vascular tone as well as a cytotoxic role as part of the ischemia associated pathology. This review discusses methods for free radical detection in brain and in other tissues, mechanisms of radical production in the course of the ischemia reperfusion process, and the efficacy of potential antioxidant agents in post ischemia therapy, especially with respect to allopurinol, an inhibitor of xanthine oxidase, and the role of taurine and its derivatives as antioxidants in different organs including the brain. PMID- 10797837 TI - [Hyperhomocysteinemia: atherothrombosis and neurotoxicity]. AB - The positive correlation existing between hyperhomocyst(e)inemia [HH(e)] and vascular disease has firmly been established through data derived from numerous epidemiological and experimental observations. Clinical data corroborate that homocysteine (Hcy) is an independent risk factor for coronary, cerebral and peripheral arterial occlusive disease or peripheral venous thrombosis. Hcy is a sulfhydryl-containing amino acid that is formed by the demethylation of methionine. It is normally catalyzed to cystathionine by cystathionine beta synthase a pyridoxal phosphate-dependent enzyme. Hcy is also remethylated to methionine by 5-methyltetrahydrofolate-Hcy methyltransferase (methionine synthase), a vitamin B12 dependent enzyme and by betaine-Hcy methyltransferase. Nutritional status such as vitamin B12, or vitamin B6, or folate deficiencies and genetic defects such as cystathionine beta-synthase or methylene-tetrahydrofolate reductase may contribute to increasing plasma homocysteine levels. The pathogenesis of Hcy-induced vascular damage may be multifactorial, including direct Hcy damage to the endothelium, stimulation of proliferation of smooth muscle cells, enhanced low-density lipoprotein peroxidation, increase of platelet aggregation, and effects on the coagulation system. Besides adverse effects on the endothelium and vessel wall, Hcy exert a toxic action on neuronal cells trough the stimulation of N-methyl-D-aspartate (NMDA) receptors. Under these conditions, neuronal damage derives from excessive calcium influx and reactive oxygen generation. This mechanism may contribute to the cognitive changes and markedly increased risk of cerebrovascular disease in children and young adults with homocystunuria. Moreover, during stroke, in hiperhomocysteinemic patients, disruption of the blood-brain barrier results in exposure of the brain to near plasma levels of Hcy. The brain is exposed to 15-50 microM H(e). Thus, the neurotoxicity of Hcy acting through the overstimulation of NMDA receptors could contribute to neuronal damage in homocystinuria and HH(e). Since HH(e) is associated with certain neurodegeneratives diseases, in the present review, the molecular mechanisms involved in neurotoxicity due to Hcy are discussed. PMID- 10797838 TI - [Physiologic role of the somatotrophic GH-IGF and others hormones in the domestic animals growth. Influence of the nutritional status]. AB - In this review main hormones involved in development and muscle growth are shown with special emphasis on growth hormone (GH) and insulin-like growth factors (IGF). Chemist composition, synthesis place, action way and main action mechanisms of these hormones are reviewed. Nutritional factors which modified seric metabolites, and their effects on hormone secretion are detailed. It was observed that GH, IGF, thyroid hormones, insulin, glucocorticoids and sexual steroids act in a complex and cordinated way to produce a productive response to different> nutritional strategies. PMID- 10797839 TI - Natural estrous cycle in normal and diabetic bitches. II). Serum nonesterified fatty acids and serum free glycerol levels during glucose and insulin tests. AB - Actions and interactions of spontaneous diabetes mellitus (DM) and natural estrous cycles (sex seasons) on the regulation of serum nonesterified fatty acids (NEFAs) and free glycerol (FG) levels in bitches in the fasting condition and during i.v. glucose (IVGTT) and insulin (ITT) tolerance tests, were studied. DM increased serum NEFAs concentration both in the basal condition and during IVGTT; it provoked a fall response to glucose load which is absent in normal controls. Estrous cycles did not modify these observations. Serum NEFAs levels during ITT were unresponsive in normal and diabetic bitches at every sex stage; flat, overlapped serum NEFAs profiles were then observed except for the diabetic group at A, which showed an early abrupt fall response of this variable from its high base line. DM increased also serum FG concentration in the fasting condition and during IVGTT. In the normal controls, serum FG base line was not affected by sex status; similarly shaped, increasing, overlapped curves during the test were observed. In the diabetic bitches "in season" (either phase), serum FG basal value was hardly above in respect to anestrous, but during IVGTT their flat profiles coincided. DM increased serum FG concentration in the basal condition and during ITT, and modified the profiles of this variable. In normal dogs in the basal condition, serum FG concentration remained unaffected by sex status; this variable hard, transiently increased during ITT, which was not influenced by "sex seasons"; therefore, similarly shaped, overlapped serum FG profiles were then observed. In the normal and diabetic bitches, serum-FG base line was not changed by "sex seasons". During ITT, serum FG mean profile in the diabetic bitches at EP was modestly above that observed in those at LP; differences for any other comparisons in normals or diabetic bitches were nonsignificant. As reported by us elsewhere, impaired glucose metabolism and absolute insulin deficiency induced ketose-prone, acidotic, insulin-dependent diabetic chryses in certain normal and diabetic beaches "in season" studied here. The unability of these animals for hydrolizing glyceride-glycerol via lipoproteinlipase (IVGTT) or via hormone sensitive fractions of lipase (ITT) and the abolished serum NEFAs suppressibility during modest hiperinsulinemia (ITT) appear to contribute to the production of such chryses. Results are discussed on the basis of interactions of serum NEFAs and FG with respective blood sugar and serum immunoreactive insulin levels as influenced by DM and estrous cycle. PMID- 10797840 TI - [Effects of the chronic malnutrition on the transferrin receptors in patients with nutritional anemia]. AB - It has been postulated that the provision of iron to the erythroid cells is under the control of a fraction of free intracellular iron. Variations in the size of this pool would send messages to either seric or cellular iron receptors aimed to keep a balance between the offer and the demand by the cells. The possibility that cell membrane receptors could be affected by chronic malnutrition was explored in these studies by the changes of iron uptake capacity of circulating mouse reticulocytes cause by the incubation of the cells with serum of either normal or anemic patients plasma donors. The results show that the incubation with nutritional anemic serum caused a significant drop in iron uptake capacity of the cells. The mechanism of the difference is discussed. PMID- 10797841 TI - Immunohistochemical study of Syntaxin-1 and SNAP-25 in the pituitaries of mouse, guinea pig and cat. AB - In the present work we have investigated the presence of the membrane proteins Syntaxin-1 and synaptosomal-associated protein (SNAP-25) by immunohistochemistry in the different parts of the pituitary of mouse, guinea pig and cat. We have demonstrated Syntaxin-1 and SNAP-25 immunoreactivity in the adenohypophysis as well as in the neurohypophysis but not in intermediate lobe. The results suggest that Syntaxin-1 and SNAP-25 are involved in the hormonal secretary process of adenohypophysis as well as neurohypophysis of these animals. PMID- 10797842 TI - Renal function in the diabetic pregnant rats. AB - Diabetic nephropathy is associated with increased urinary albumin and reduce kallikrein excretion. Increased activity of the renal kallikrein-kinin system has been suggested as one of the possible mechanisms underlying diabetic hyperfiltration. The present study shown that the Kallikrein-kinin system is progressively increased in the diabetic-pregnant rats at 7, 14, 21 days; 48 and 7 days after pregnancy (P < 0.05 vs Control). However, this increase during diabetic pregnancy did not reached the levels of control pregnancy. On the other hand albumin excretion shown a significant and progressive renal damage in the diabetic state. These findings suggest that the diabetic pregnancy could impair the renal hemodynamic, but, on the other side could modulate the vasodilator system at pregnancy in the attempt to protect the fetus. PMID- 10797843 TI - Influence of anterodorsal thalamic nuclei on the hypophyseal-adrenal axis and cardiac beta receptors in rats submitted to variable chronic stress. AB - The limbic structures play an important role in the control of the neuroendocrine and sympathical adrenal function in basal and stress conditions. This work was undertaken to evaluate plasma ACTH, adrenocortical activity, cardiac adrenoceptors density and affinity response to variable chronic stress (VCS) in anterodorsal thalamic nuclei (ADTN) lesioned rats. Thirty days after lesion, shamlesioned stressed animals increased plasma ACTH and corticosterone as compared to sham-lesioned unstressed animals (p < 0.05); lesioned rats increased ACTH levels after VCS (p < 0.05) as compared to unstressed-lesioned rats. Whereas in sham-lesion plasma corticosterone (C) increased after stress, in lesioned animals(C) remained unchanged as compared to unstressed-lesioned animals. In the stressed groups, adrenal C contents were below those found in unstressed rats. beta-receptors affinity, in all the experimental groups, was similar, but VCS sham-lesioned animals underwent a significant increase in cardiac D-adrenergic receptors density when compared with basal and lesioned groups (P < 0.001). Our findings would demonstrate that the increment in cardiac beta adrenoceptors density appears as a consequence of the increase in ACTH, plasma corticosterone and sympathetic response provoked by chronic stress situations. ADTN lesion attenuated this hipophisoadrenal system response to chronic stress as well as the above mentioned cardiac beta adrenoceptors density increment. PMID- 10797844 TI - Regression with repeated measures in the experimental units. AB - The most satisfactory solution to the problem of modeling a family of regressions with repeated measures in the experimental units is multivariate in nature. However, multivariate methods are difficult to follow and implement. Furthermore, by keeping the focus on the experimental unit, a family of simple univariate linear models will often parallel both the investigator's intuitive grasp of the statistical task at hand. We present two examples based on data from a study of the suckling stimulus during breastfeeding in newborn infants. We show how a family of regression lines can provide useful, if approximate, answers to the questions of interest. One example involves a regression setting proper and the other a typical case of correlation. We discuss alternative univariate models that may be useful for this type of problems. PMID- 10797845 TI - Myocardial contractile response to an oral glucose load in normal subjects evaluated by echocardiography. AB - There is a paucity of experimental data on the actual mechanism of insulin induced changes on the myocardial function. In the present study we investigated the myocardial contractile, response to an oral glucose load using echocardiography. Fifteen healthy volunteers were studied after overnight fast and 150 minutes after the oral load of 75 g glucose. Oral glucose load caused an increase in plasma glucose and insulin levels, which was accompanied by a significant increase in left ventricular shortening (from 35.2 +/- 0.7% at baseline, to 38.5 +/- 0.6% and 39 +/- 0.9% at 30 and 60 minutes post glucose load, respectively [P < 0.05 vs baseline]; ejection fraction rose from 0.73% +/- 0.01 to 0.77% +/- 0.01 (P < 0.05); pressure rate product increased from 7.29 +/- 0.2 to 8.31 +/- 0.3 mmHg x beats per min (P < 0.007) and heart rate enhanced from 68.3 +/- 1.9 to 74 +/- 1.6 (P < 0.034) and 75.3 +/- 1.5 beats per min (P < 0.008) at 60 and 90 minutes after glucose, respectively. Meanwhile, mean arterial pressure decreased significantly (10 +/- 1.5%, P < 0.018) when compared to basal values. These results indicate a significant change in the myocardial contractile response to an oral glucose load, probably related to baroreceptor reflex response as well as an overridden by a potent vasodilator action of insulin. Nevertheless, we could not rule out that the cardiac effects may also be due an insulin-induced sympathetic activation or a direct myocardial effect. PMID- 10797846 TI - Lp(a) and LDL2: differences in fatty acid composition and collagen lattice contraction activity. AB - Lp(a) and LDL2 were used for detailed fatty acid analyses and tested in an in vitro model for promotion of fibroblast-mediated collagen lattice contraction to determine possible compositional and functional differences between these two apoB-containing lipoprotein species. Autologous Lp(a) was more saturated with respect to fatty acid composition than LDL2 in triglyceride and cholesterol ester lipid classes and had differences in the fatty acid content of phospholipids. Functionally, LDL2 promoted rapid fibroblast-mediated contraction while Lp(a) was significantly less active in promoting rapid contraction on a protein per weight basis. These studies suggest a synthetic route for Lp(a) diverging from the majority of other apoB-containing lipoproteins and significant activity of LDL2 in a collagen lattice contraction system. PMID- 10797847 TI - Stable isotope techniques for the detection of Helicobacter pylori infection in clinical practice. 13C-Urea Breath Test in different experimental conditions. AB - The 13C-UBT has been demonstrated to be a reliable method for the evaluation of Helicobacter pylori infection. The aim of our work is to determine the cut-off point of the 13C-UBT for samples collected as gas or collected in a solution of triethanolamine. For this purpose, patients fasted for at least 6 hours were able to collect basal samples before the administration of 65 mg of 13C-urea solution. Breath samples were taken 10, 30 and 60 minutes after the administration of the labeled solution. All the samples were collected in gas collectors and in glass vials containing 1 ml of a 7% triethanolamine solution. The cut-off points for gas collected samples were established in 4.0/1000, 4.6/1000 and 4.4/1000 for 10, 30 and 60 minutes samples, respectively, while for the samples collected in triethanolamine solution, cut-off points were established in 5.0/1000, for the 10 minutes samples, in 3.5/1000 for the 30 minutes samples and 4.7/1000 for the 60 minutes samples. We found that this test has a sensitivity of 100% and a specificity of 100% for H. pylori detection in both experimental conditions, when multiple breath samples are taken. If we considered only the 30 minutes time, sensitivity and specificity diminish for the gas collected samples. We conclude that the collection of breath samples in triethanolamine solution allows a better differentiation between H. pylori infected and non infected patients than gas collected samples. PMID- 10797848 TI - Enzymatic method for branched chain alpha-ketoacid determination: application to rapid analysis of urine and plasma samples from maple syrup urine disease patients. AB - A new method for the determination of branched-chain alpha-ketoacid concentration using lactate dehydrogenase (E C 1.1.1.27) isozyme C4 (LDH C4) from mouse testes is proposed. The assay is performed on urine and plasma without previous treatment. Alpha-ketoglutarate and pyruvate are determined on the same sample using glutamate dehydrogenase (GDH, EC 1.4.1.2) and lactate dehydrogenase isozyme A4 (LDH5) respectively and subtracted from the total alpha-ketoacid concentration obtained with LDH C4. This value corresponds to the branched chain alpha ketoacid. Results were linear within the concentration range 8 to 170 mumoles/L. Detection limit was 8 mumoles/L. Analytical recovery was higher than 91%. For microplate assays, recoveries were higher than 84% and the detection limit was 20 mumoles/L. Determinations performed with GDH, LDH A4 and LDH C4 allow differentiation of E3 deficiency from other clinical phenotypes of maple syrup urine disease. The method is simple and fast, and adaptation to microplates would allow screening of newborns. PMID- 10797849 TI - The effects of calcium channel blocker, flunarizine on spinal reflexes in the cats. AB - Effects of calcium channel blocker flunarizine on spinal monosynaptic reflexes were investigated in spinal cats. Flunarizine was administered locally into the spinal cord (10, 50, 100 microM) and intraperitoneally (5, 10, 20 mg/kg). Adult cats (n = 10), weighing 1.5-3 kg were anesthetized with ketamine (50 mg/kg, i.m.) and artificially ventilated. Animals were spinalized at C1 level. A laminectomy was performed in the lumbosacral region. The ventral and dorsal roots of segment L5 were isolated and a pouch of skin was formed at the site of the dissection to allow the exposed tissues to be covered with liquid paraffin. The temperature was kept at 38.5 degrees C with a heating pad. A polyethylene cannula was introduced into the left carotid artery to monitor blood pressure, which was kept above 100 mmHg. The dorsal root of segment L5 was placed on a silver-silver chloride wire electrode for stimulation through an isolation unit. The reflex potentials were recorded from the ipsilateral L5 ventral root, mounted on a silver-silver chloride wire electrode. The systemic (5, 10, 20 mg/kg) and local (50 and 100 microM) dosages of cinnarizin derivative flunarizine significantly decreased the amplitude of reflex response (p < 0.05). Moreover, the latency of the monosynaptic reflexes was increased after administration of the drug (p < 0.05). Voltage-dependent calcium channels in the spinal cord may play an important role to regulate reflex respond. PMID- 10797850 TI - Effect of methyl-prednisone and cyclosporine on the lipid pattern and polyunsaturated fatty acid biosynthesis in the rat. AB - The hyperlipidemia posttransplant has been largely attributed to immunosuppressant agents. In the present work we evaluated the effect of oral administration of cyclosporine (5 mg/kg/day) and/or methyl-prednisone (1 mg/kg/day) on lipid composition and polyunsaturated fatty acid biosynthesis in normal adult male rats. The results obtained showed that both agents produced a delay on the growth together with a significant loss of body weight. In liver microsomal fraction from rats treated with methyl-prednisone, a depression in delta 6 and delta 5 desaturation activities, was observed. This effect was corroborated in the fatty acid pattern through the enhancement of linoleic and dihomo-gamma-linolenic acids, and a depression of arachidonic acid. Similar results were noticed in those rats treated with both drugs when compared to the controls. No changes were observed either in the amount of liver microsomal total lipids or in the fatty acid composition of kidney and testis microsomes, as well as in erythrocyte membranes, among the different groups studied. Cyclosporine alone produced a significant depression in plasma triglycerides and showed no modifications in the other lipid parameters studied compared to the controls. Fluorescence anisotropy measured in the different membranes was not modified by the several treatments used. In view of the aforementioned data, it can be stated that methyl-prednisone would be the responsible for many of the lipid disorders that can be observed in posttransplant patients when they are subjected to the combined immunotherapy with cyclosporine. PMID- 10797851 TI - Effect of calcium deficiency on acyl-CoA synthetase activity in microsomes from various rat tissues. AB - The specific activity of long-chain acyl-CoA synthetase in microsomes from various tissues of control and calcium-deficient rats was determined. It was found that the saturated acids, palmitic and stearic, were preferential substrates compared to the non-saturated linoleic, alpha-linolenic and eicosa 8,11,14-trienoic acids. All of them showed similar Vm values with different affinity constants. After 60-day treatment on a calcium-deficient diet (0.5 g Ca/Kg diet), a significant increase in the acyl-CoA synthetase activity was observed for all the tested fatty acids in liver and kidney microsomes. These changes were evoked without any modification in the substrate selectivities shown for the control microsomes, and they were well-correlated with calcium level in both tissues. Under the calcium deficient state an increase in Vm values was observed for palmitic and eicosatrienoic acids with no changes in the corresponding Km, suggesting an increment in the number of active enzyme molecules within the microsomal membrane. PMID- 10797853 TI - [Hemorrhagic, edema-forming, proteolytic and myonecrotic activities of viper venoms of Bothrops alternatus (Vibora de la Cruz)]. AB - A study of hemorrhagic, edema-forming, proteolytic and myonecrotic activities induced by newborn Bothrops alternatus snake venoms from Argentina was performed. The analyzed venom had these activities with markedly differences when compared with the venom of adult specimens. Hemorrhagic activity was four times higher, while edema forming activity was ten times or more. But proteolytic activity was similar in both groups. Hystological analysis showed intense hemorrhage and muscular fiber myolisis after 60 minutes of venom injection. Myonecrosis and inflammatory exudate were higher in the following hours. Hemorrhage decreased after 24 hours and muscular fiber regeneration started after the first week with granulation tissue formation. On the fourth week regenerating muscular fiber areas and granulation tissue were still observed. As a conclusion, we can assume that the injury induced in the snake accident by newborn Bothrops alternatus snakes would be more intense than those induced by the adult ones. PMID- 10797852 TI - Influence of moderate cooling (37 degrees C-25 degrees C) on the reactivity of isolated rat tail artery. AB - The aim of the investigation was to examine the effects of cooling on the tail artery regarding the scarceness of such studies in spite of the essential thermoregulatory role played by this vessel. Segments of the proximal portion were suspended isometrically in medium containing 1.25 mM Ca. Lowering the temperature to 25 degrees C increased the sensitivity and maximum strength of the adrenaline concentration-effect curves. These changes were reversed by warming back to 37 degrees C. Cocaine attenuated the increase of sensitivity without changing the increase of the maximum response. Either the sensitivity and strength of the responses to phenylephrine and serotonin were increased by cooling. Clonidine evoked weak contractions in 18 out of 38 experiments. After cooling, the responses persisted only in 7 arteries and the strength was almost halved. Responses to field electric stimulation at 25 degrees C exhibited a pronounced increase of strength and a small increase of sensitivity. -log Kb for prazosin against adrenaline was increased by cooling (8.7 and 9.1 at 37 degrees C and 25 degrees C, P < 0.01). After partial receptor inactivation using phenoxybenzamine, the dissociation-constant (KA) indicated a moderate affinity for phenylephrine that was not changed by cooling (4.1 and 4.2 x 10(-6) at 37 degrees and 25 degrees C respectively). Receptor reserve and occupancy at EC50 also remained unchanged at 25 degrees C. It can be concluded that: 1) cooling increases the tail artery reactivity, partly as a consequence of the inhibition of adrenergic neuronal uptake; 2) responsiveness to alpha 2-agonists is not involved in the effects of cooling whereas the role of alpha 1-adrenoceptor could not be properly clarified; 3) cooling may facilitate some steps of the contractile activation beyond the agonist-receptor interaction. PMID- 10797854 TI - Effects of oxytocin and an oxytocin receptor antagonist on retention of a nose poke habituation response in mice. AB - The present study describes the use of nose-poke habituation as a memory task in mice and demonstrates that it is sensitive to oxytocin (OT) and an oxytocin receptor antagonist (AOT) administered after the learning trial. Habituation of nose-poke behavior of mice was defined as a reduction in number of nose-pokes compared to baseline, and was measured in a hole-board apparatus to which male Swiss mice were exposed on two consecutive days for 5 min, respectively. Immediate post-training subcutaneous administration of OT (2.00 micrograms/kg) impaired retention performance, whereas AOT (0.20 microgram/kg) enhanced it. Neither the impairing effects of OT (2.00 micrograms/kg) nor the enhancing effects of AOT (0.20 microgram/kg) were seen when the training treatment interval was 180 min, suggesting that both treatments influenced the storage of recently acquired information. The effects of OT (2.00 micrograms/kg) on retention were prevented by AOT (0.02 microgram/kg) administered immediately after training but 10 min prior OT treatment. This dose of antagonist did not affect retention by itself which suggest that impairing effects of OT on retention are probably due to an interaction of the neuropeptide with specific receptors. The actions of OT and AOT on retention were not due to enduring proactive effects of the compounds on performance during the retention test, since when given to untrained mice did not modify their spontaneous activities in the hole-board when recorded 24 h later. We suggest that nose-poke habituation learning can be a suitable method to investigate the mnestic effects of drugs, and that oxytocin negatively modulates memory storage of this form of learning elicited by stimuli repeatedly presented without reinforcement. PMID- 10797855 TI - Antispasmodic action of propinox on the isolated human gallbladder: possible mechanism of action. AB - Propinox is an antispasmodic drug frequently used in the treatment of disorders of the gastrointestinal tract, the uterus and the gallbladder, but little is known about its relaxing activity in gallbladder tissue. The main objective of this study was to determine the antispasmodic activity of propinox, compared to other antispasmodics, in the gallbladder and to assess its binding affinity to receptor sites which may be involved in its mechanism of action. Antispasmodic activity of propinox, (-) scopolamine-n-butyl bromide, atropine and verapamil was determined in human gallbladders to reduce the risk of interspecies variability. Inhibitory activities (ED50) of carbachol-induced contraction were: atropine 5.03 x 10(-8) M > propinox 1.25 x 10(-7) M > verapamil 6.63 x 10(-6) M > (-) scopolamine-n-butyl bromide 5.4 x 10(-5) M. pD'2 for propinox was 6.94, indicating non competitive inhibition of carbachol action. Radioligand binding studies were performed to determine if the antispasmodic action of the drug involved binding to muscarinic receptors or calciumantagonist sites. The inhibition constant (Ki) of propinox for muscarinic receptors of guinea pig ileum smooth muscle, which contains a mixed M2-M3 receptor population, was 1.6 x 10(-6) M. Ki for brain muscarinic receptors (M1) was 1.0 x 10(-4) M, for cardiac receptors (M2) 1.2 x 10(-6) M and from salivary gland receptors (M3) 1.5 x 10(-6) M. For binding to the dihidropiridine calcium antagonist binding sites, Ki were: 4.9 x 10(-5) M for propinox and 2.2 x 10(-7) M for verapamil. For the phenylalkylamine binding sites Ki were: 5.0 x 10(-6) M for propinox and 3.5 x 10( 8) M for verapamil. For the benzothiacepine binding sites, Ki for propinox was 5.2 x 10(-6) M. The following may be concluded: 1.--The antispasmodic activity of propinox in isolated human gallbladder was comparatively less potent than that of atropine and more potent than those of verapamil and (-) scopolamine-n-butyl bromide. 2.--Propinox showed binding to muscarinic and calcium receptors that can be related to its antispasmodic activity; suggesting that the drug is an antispasmodic with anticholinergic and musculotropic activity. 3.--The dual mechanism of action, anticholinergic and calcium-blocking, would induce synergism of pharmacodynamic effects and minimize adverse events of pure antimuscarinic drugs or calcium antagonists. PMID- 10797856 TI - [Experimental model for the study of molybdenosis in the primary copper deficiency in rats]. AB - An experimental model in rats was evaluated to differentiate the effects between Copper deficiency and Molybdenosis. Sixty weaning rats (30 male and 30 female) received a diet with 70% complete powder milk (1 ppm Cu) and 30% maize meal (0.8 1.5 ppm Cu). Three experimental groups received the following mineral supplementation: copper deficiency (40 ppm Fe), molybdenosis (40 ppm Fe + 40 ppm Cu + 500 ppm Mo) and control (40 ppm Fe + 40 ppm Cu). The animals were weighed each 14 days. At 70 days of treatment were sacrificed. Blood and liver were sampled for analyzing hematocrit, ceruloplasmin activity and Cu and Mo liver concentration. Copper deficiency group had less serum ceruloplasmin activity. Cu and Mo liver concentration were higher in the animals with molybdenosis. We concluded that when Cu levels are higher than minimum requirement, feeding with high Mo, do not affect ceruloplasmin activity. In addition, high Mo liver concentration allows us to elucidate effects "per se" of molybdenosis. PMID- 10797858 TI - Investigation of serum calcium, phosphorus, albumine uric acid and lipid parameters in sportsmen and sedanters. AB - In this study, it was aimed to investigate the differences in blood lipid fractions which are important risk factors for atherosclerosis and to show how the levels of serum albumin, uric acid, ionized calcium, inorganic phosphorus change with exercises between sportsmen and sedanters. 25 sportsmen and 25 sedanters (13 male, 12 female in each group) aged 18-23 were chosen for the study. From the venous blood samples taken from the two groups, levels of serum total lipid, total cholesterol, HDL-cholesterol, LDL-cholesterol, VLDL cholesterol, triglyceride, albumin, uric acid, ionized calcium, inorganic phosphorus were analyzed. Student's t-test was used for the statistical analyses. There were some important changes in the levels of serum triglyceride, VLDL cholesterol and ionized calcium of sportsmen compared to those of controls. PMID- 10797857 TI - [Toxic and enzymatic activities of the Bothrops moojeni venom from Argentina]. AB - Toxic and enzymatic activity of Bothrops moojeni venom from Argentine. Hemorrhagic, edema forming, proteolytic and indirect hemolytic activities, myonecrosis and muscle regeneration using optic microscopy induced by Bothrops moojeni snake venom from Argentina were studied. The described activities were present in this venom. Myonecrosis was intense 60 minutes after venom injection, increasing its activity during the first 3 hours. Myolytic lesions rise above coagulative ones. Hemorrhage was minimum the same as the edema. After 3 hours polymorphonuclear inflammatory infiltrate was observed, with neutrophils. Macrophages were visible after 48 hours. After 3 days fibroblasts were present. At the 7 day muscle fiber regeneration were present, showing striations alternating with calcified areas. Bothrops moojeni venom injuries are intense and the recuperation of them can generate important tissue lose, with functional consequences. PMID- 10797859 TI - Erythropoietin assay in mice made polycythemic by transfusion of heterologous red cells. AB - A simple in vivo bioassay suitable for routine testing of quality control of recombinant human erythropoietin (rHu-EPO) analogues was developed. Mice made polycythemic by intraperitoneal injection of 1.2 ml of a 80% suspension of heterologous (rat) red cells were used as assay animals and splenic 59Fe uptake as expression of the response to rHu-EPO. The assay took three days and the following schedule is proposed: 1) intraperitoneal injection of 1.2 ml of washed packed red cells obtained from donor rats, 2) subcutaneous injection of test material 4-5 h after transfusion, 3) intravenous administration of 59Fe tracer 48 h later, and 4) determination of splenic isotope uptake 6 h after injection. This method for the in vivo bioassay of rHu-EPO analogues is an economical and reliable alternative to the existing bioassays of the hormone. PMID- 10797861 TI - Effects of intrathecal L- and N-type calcium channel blockers on the antinociception evoked by opioid agonists in the rat tail flick test. AB - The effects of intrathecal administration of nimodipine or omega-conotoxin GVIA (L- and N-type calcium channel blockers, respectively) alone or followed by DAMGO, DADLE or bremazocine (mu-, delta- and kappa-opioid agonists, respectively) were studied on the rat tail flick test. The N- (16 to 64 pmoles), but not the L type blocker (60 to 240 pmoles) produced a dose and time-dependent increase in the latency for the tail-flick reflex. DAMGO (30 to 120 pmoles) or bremazocine (190 to 560 pmoles), but not DADLE (50 to 200 pmoles), produced a dose-dependent increase in the latency for the tail-flick reflex. The effect of the highest dose of DAMGO was smaller, while the effects of DADLE and bremazocine were not changed after nimodipine (60 pmoles). The effects of DADLE were significantly potentiated, while the effects of DAMGO and bremazocine were not changed after omega-conotoxin GVIA (16 pmoles). The intrathecal administration of an N-type calcium channel blocker with a delta-opioid agonist seems to be the most effective combination to produce antinociception in the rat tail flick test. PMID- 10797862 TI - Hair cell loss and regeneration in the chick cochlea after treatment with gentamicin. AB - Hair cells can be damaged by countless agents, among them aminoglycoside antibiotics. In the chick cochlea, the hair cell loss can be recovered by regeneration. The objectives of the present investigation were to study the time progression of injury caused by gentamicin and the regeneration process in chick hair cells. Gentamicin was administered in a single subcutaneous dose of 125 or 250 mg/kg to two groups of 3-day old chicks. The cochleae were processed for analysis by scanning electron microscopy on the 1st, 3rd, 5th and 20th day after injection. The cellular sequence of degeneration and regeneration was studied. On the 20th day, most it he damaged cochlear area showed regenerated hair and support cells. Stereocilia and microvilli were observed on the apical surface of the regenerated hair cells. PMID- 10797860 TI - In honour of the 70th birthday of Professor Alexandre Pinto-Corrado, from the School of Medicine of Riberao Preto, University of Sao Paulo (FMR/USP) in the occasion of his retirement. PMID- 10797863 TI - The serotonin paradox: drug-receptor interaction in rat vas deferens. AB - The contractile effect of serotonin was studied in rat vas deferens, in comparison with that of noradrenaline and tyramine, after reserpine treatment, surgical denervation, and transplantation to the colon. In reserpinized animals the effect of 5HT resembled that of tyramine, since it was strikingly reduced, in spite of a small residual effect, showing that in normal preparations the effects of 5HT and tyramine are predominantly due to the release of endogenous noradrenaline. However, in denervated or transplanted vas deferens, in which the effect of tyramine is also abolished, the effect of 5HT was potentiated. It is suggested that after chronic, long lasting depletion of endogenous noradrenaline, there are alternate mechanisms that are generated to improve the contractile effect of 5HT, but not of tyramine. The nature of these mechanisms is still unknown. PMID- 10797864 TI - Modulation of acetylcholine release by presynaptic muscarinic autoreceptors. AB - The existence of a modulatory system controlling the acetylcholine (ACh) release was first proposed for the nicotinic subtype in 1962. Following the first observation of a possible positive feedback loop activated by the released Ach, many studies were oriented in the investigation of the involved presynaptic autoreceptors. Most of the data have been obtained at the motor end-plate, commonly defined as the simplest model of peripheral synapse. The characterization of the chemical transmission since its first proposal showed a more complex pattern involving both the cholinergic and the adrenergic systems. It is now evident that this regulation is widespread both in the central and in the peripheral nervous system. The evidence that the release of ACh can be up- or down-regulated by the transmitter itself (autoregulation) or other neuromediators (heteroregulation) is now proved. In the last decades the attention was focused to the identification of the receptor subtypes located on the releasing nerve terminal. For the purpose, different techniques were used in the various laboratories. The functional approach was based mainly on the electrophysiological characterization of the events evolved prior, during and after the activation of the motor endplate nicotinic receptor. On the other hand, the overflow studies were carried out using radiolabeled ACh (rACh) obtained treating muscle fibers with radioactive choline (rCh). Many scientific papers proposed common data indicating a clear positive (nicotinic) or negative (muscarinic) modulation of the ACh release. Temporally, the description of the muscarinic regulation followed the discovery of the nicotinic one. However, by a pure pharmacological point of view it represents a challenge due to the more complex organization and function. In the peripheral nervous system, i.e. neuromuscular, the meaning of both the muscarinic and nicotinic modulations may appear as free of function. Conversely, in the central systems some effects, such as antinociception and others, could represent the basis of a functional activity such as proposed by Corrado group. The complete characterization of this phenomenon by a physiological and a pharmacological point of view could represents the goal for future uses and therapeutic potential. The present review illustrates the know how and the efforts in the characterisation of the muscarinic regulation of transmitter release from the beginning of its discovery trying to order the numerous scientific data published in this field. Furthermore, our personal data obtained with the Loose Patch Clamp (LPC) technique will be briefly presented and discussed. Our work was built up using agonists and antagonists of the muscarinic receptor subtype in the aim of better characterize the modulation function of the mediator Ach. We used carbachol (Cch), oxotremorine (Oxo) and dl-muscarine as agonists and 1-hyoscyamine, pirenzepine, ipratropium, 11[[2-1[(diethylamino) methyl-1-piperidinyl]-acetyl]-5, 11-dihydro-6H-pyrido [2,3-b][1,4] benzodiazepine-6-one (AFDX-116), methoctramine and 1,1-dimethyl-4 diphenylacetoxy-N-methylpiperidine (4-DAMP) as antagonists. PMID- 10797865 TI - Mechanisms of the UTP-induced tension in mammalian skeletal muscles. AB - The mechanisms of UTP-induced tension in human and rat skinned fibers were investigated using isometric tension recordings, electrophysiological techniques and biochemical methods. In fast-type fibers from rat extensor digitorum longus (EDL) the UTP-induced tension: a) required previous loading of Ca2+ into the sarcoplasmic reticulum (SR); b) was inhibited by previous exposure to caffeine; c) was abolished by functional disruption of the SR; d) was not affected by blockade of the SR Ca(2+)-release channels by ruthenium red or heparin; e) was prevented by spermidine. These data point to the SR as the target of UTP action and suggest a pathway of UTP-induced Ca(2+)-release independent of the ryanodine- or the IP3-sensitive Ca(2+)-release channels. Accordingly, UTP failed to stimulate the electrophysiological activity of ryanodine-sensitive channels, incorporated into lipid bilayers. We suggest that UTP-induced Ca(2+)-release might occur via the channel form of the SR Ca(2+)-ATPase. The UTP-induced tension in human slow-type fibers was not affected by the SR Ca2+ content or by disruption of the SR, but was accompanied by changes in the tension-pCa relationship, namely increase in maximum Ca(2+)-activated tension, and in apparent Ca(2+)-affinity of troponin. The UTP-induced tension in slow-type fibers from rat soleus was partially inhibited by Ca(2+)-depletion from, or by disruption of the SR, and was accompanied by changes in tension/pCa relationship, similar to those observed in human fibers. Both in skinned fibers and in isolated SR vesicles, UTP was less effective than ATP as a substrate for the SR Ca(2+) ATPase. This effect might contribute to the UTP-induced tension. PMID- 10797866 TI - Bioequivalence evaluation of two roxithromycin formulations in healthy human volunteers by high performance liquid cromatography coupled to tandem mass spectrometry. AB - The bioequivalence of two different formulations containing roxithromycin (SPE 712-1). Oral suspension 300 mg/15 mL as test formulation and Rotram, tablets 300 mg as reference formulation, both by Schering Plough S.A., Brazil) was evaluated in 24 healthy volunteers of both sexes (12 male and 12 female). The study was conducted open with randomized two-period crossover design and a 14-day washout period. Each subject received 300 mg of each roxithromycin formulation. Plasma samples were obtained over a 72-hour interval and roxithromycin concentrations were analyzed by combined LC-MS/MS with positive ion electrospray ionization using selected ion monitoring method. From the plasma roxithromycin concentration vs time curves the following pharmacokinetic parameters were obtained: AUC(0-72 h), AUC(0-infinity), Cmax, t1/2 ratios and tmax individual differences. The 90% for confidence interval (CI) of geometric mean SPE-712-L/Rotram individual percent ratio were 105.0-128.3% for AUC(0-72 h), and 78.4-96.9 for Cmax. Although this 90% CI were marginally outside the interval proposed by the Food and Drug Administration, the probability assessed by the two-one sided West for ratios was included in the 0.8-1.25 interval, as we concluded that SPE-712-L oral suspension formulation was bioequivalent to Rotram tablet formulation for the extent and rate of absorption. PMID- 10797868 TI - Biochemistry and pharmacology of aminoglycoside-induced hearing loss. AB - Discovered in the 1940s, the aminoglycoside antibiotics were the long-sought remedy for tuberculosis and other serious bacterial infections. They are still one of the most commonly used antibiotics today, thanks to the combination of their high efficacy with low cost. This review begins by addressing some of the history and the acute side effects of aminoglycosides. It then details their chronic toxicity to the inner ear (ototoxicity). The review concludes with recent advances that have demonstrated free-radical reactions as the underlying mechanism and intervention with antioxidant/chelation therapy to prevent aminoglycoside ototoxicity. PMID- 10797867 TI - Sites and mechanisms of antibiotic-induced neuromuscular block: a pharmacological analysis using quantal content, voltage clamped end-plate currents and single channel analysis. AB - Since the original observation of Vital Brazil and Corrado (1957) concerning the antibiotic induced neuromuscular block produced by streptomycin, there has been considerable interest in the mechanisms responsible for not only neuromuscular block but also the effects of antibiotics on different systems. We used the voltage clamped end-plate of transacted skeletal muscle to examine the concentration-dependent actions of several groups of antibiotics. The aminoglycoside antibiotics, neomycin and streptomycin, were both more effective at reducing quantal release of acetylcholine (ACh) than interacting with the postjunctional ACh receptor-channel complex. Neomycin was approximately 10 X more potent prejunctionally than streptomycin and the prejunctional effects of each antibiotic were reversed competitively by raising extracellular calcium. Both neomycin and streptomycin also had postjunctional actions at higher concentrations. Neomycin interacted with the open state of the ACh receptor ion channel complex while streptomycin blocks the ACh receptor. The lincosamide antibiotics, lincomycin and clindamycin produced their neuromuscular block postjunctionally by interacting with the open state of the ACh-receptor channel complex. Clindamycin is approximately 20 X more effective at blocking the open channel than was lincomycin. Using cell attached patch clamp recordings in cultured rat myotubes, we demonstrated a lincosamide-induced block of open ion channels with clindamycin having a much slower unblocking rate than lincomycin. Using epimers of the lincosamides, we demonstrated that lipophilicity of the molecule, rather than stereochemical considerations, is important for open channel blockade affecting primarily the "off" rate of channel blocking. This mechanism appears important for not only the lincosamide antibiotics but also for the postjunctional actions of the aminoglycoside antibiotics, particularly neomycin. PMID- 10797869 TI - A review on conotoxins targeting ion channels and acetylcholine receptors of the vertebrate neuromuscular junction. AB - In this article we present an overview of some peptides extracted and purified from the venom of marine snails of the genus Conus. These active peptides named conotoxins can be used as research tools to target voltage-gated ion channels as well as ligand-gated receptors. Because of their relatively small size, conotoxins can be chemically synthesized and made widely available. In this review we focus on conotoxins that target voltage-sensitive sodium channels, voltage-dependent calcium channels and nicotinic acetylcholine receptors of the vertebrate neuromuscular junction. Emphasis is given on summarizing our current knowledge of their primary structure and their specific pharmacological actions at the pre- and the post-synaptic level of the neuromuscular junction. Evidence is presented for conotoxins that discriminate between pre- and post-synaptic voltage-gated sodium channels. Among these peptides, the mu-conotoxin family is well characterized by its ability to block selectively sodium channels in skeletal muscle fibres without affecting axonal and nerve terminal Na+ channels. Furthermore, new conotoxins like Conus consors toxin (CcTx) and conotoxin EVIA selectively target Na+ channels in axons and nerve terminals without affecting skeletal muscle fibres. omega-conotoxins known as highly potent and selective blockers of voltage-sensitive calcium channels have proven to be valuable in determining the roles of the various subtypes of channels involved in acetylcholine release from motor nerve endings. Finally, Conus peptides which act at muscle nicotinic acetylcholine receptors constitute the most extensive characterized family of conopeptides that exhibit sequence similarity, different structural motifs and surprising diversity in their competitive and non competitive actions. PMID- 10797870 TI - Natural probes for cholinergic sites: L-bebeerine actions on the neuromuscular transmission, the nicotinic receptor/ionic channel complex, and contraction of skeletal muscles. AB - The mechanisms underlying the muscle relaxant activity of 1-bebeerine (BB), a tertiary alkaloid isolated from the roots of Chondrodendron platyphyllum, were examined in mammalian and amphibian skeletal muscles. Injections of BB (0.05-1 g/kg, i.p.) in rats caused a dose-related flaccid paralysis and respiratory arrest at high doses. In isolated rat diaphragm and toad sartorius muscles, BB depressed the indirectly elicited muscle twitches (IC50: 228 microM and 5.4 microM, respectively, at 22 degrees C) and blocked the nerve-elicited muscle action potential. The neuromuscular blockade was not reversed by neostigmine (10 microM). High concentrations of BB (170 and 340 microM) caused muscle contracture unrelated to the junctional blockade, and intensified by increasing the bath temperature. Analysis of the contraction properties showed that BB (40 and 80 microM) increased the twitch/tetanus ratio (46% and 125%) and prolonged the relaxation time; the falling phase of the directly elicited action potential in toad sartorius muscle fibers was slower probably by a decreased potassium conductance. BB (0.1-340 microM) reduced the binding of [125l]alpha--bungarotoxin to the junctional ACh receptor of the rat diaphragm (IC50: 47.7 microM, at 37 degrees C. At low concentrations BB (1.5-15 microM) induced either opening or blockade of the ACh receptor-ionic channel. The results showed that BB blocked noncompetitively the neuromuscular transmission through a mechanism that affects the ACh recognition site and the ionic channel properties. The alkaloid also produced muscle contracture and changed the contractile properties through its extra-junctional action at the calcium handling by the sarcoplasmic reticulum or the contractile machinery. PMID- 10797871 TI - Effect of electrolytic lesion of the median raphe nucleus on behavioral and physiological measures of stress. AB - The effect of electrolytic lesion of the median raphe nucleus was measured on behavioral and physiological parameters related to stress 24 h after the lesion. In of the elevated plus-maze the lesion decreased the percentage of open arm entries and tended to shorten the time spent on the open arms indicating an increase in anxiety. In contrast, the lesion markedly increased the time spent in the bright (aversive) compartment of the light-dark box and decrease in attempts to cross from the dark toward the bright compartment, an anxiolyic effect. With the exception of plasma prolactin level, which was lowered by the lesion, the physiological measures used in the present study indicate that the lesioned animals are under stress. Thus, death rate and weight loss after the surgery were higher in lesioned than in control animals. In addition, lesioned animals showed higher plasma corticosterone levels, a high incidence of gastric ulcers in the fundus and a depressed immune response to the mitogen concavaline A. These results highlight the importance of the median raphe nucleus in the regulation of stress and anxiety. They also show that behavioral and physiological measures of stress may be dissociated. PMID- 10797872 TI - Electrophysiological and ultrastructural analysis of the neuromuscular blockade and miotoxicity induced by the Micrurus nigrocinctus snake venom. AB - Micrurus nigrocinctus is the most abundant coral snake in Central America. The venom of this specie induced a concentration-dependent (10-20 micrograms/ml) depolarization in the isolated mouse phrenic nerve-diaphragm preparations incubated at 37 degrees C. d-Tubocurarine (10 micrograms/ml) and (alpha beta ungarotoxin (3-5 micrograms/ml) were able to partially protect against the depolarization induced by the venom (10 micrograms/ml), suggesting the involvement of subsynaptic cholinergic receptors. This venom (10 micrograms/ml) also increased the frequency and amplitude of miniature end-plate potentials (mepps) during the first 10-20 min of incubation. Subsequently, the mepps progressively decreased and disappeared after 60 min. These responses were accompanied by ultrastructural changes involving the nerve terminals, the subsynaptic junctional folds and the muscle mitochondria. The synaptic gutter was shallow and, very often, "shrunken" terminals with omega-shaped axolemmal indentations and a decreased number of synaptic vesicles were present. A common finding was the presence of numerous finger-like, membrane-bounded bodies interposed between the terminal and the Schwann cells or postsynaptic sarcolemma. The preincubation of the venom with specific antivenom or the incubation of the preparations at room temperature (24-26 degrees C) reduced the number and intensity of the ultrastructural alterations. The last finding suggests the involvement of a enzymatic process, probably a phospholipase A2, present in the venom. There was a good correlation between the electrophysiological and ultrastructural effects induced by the venom which allow us to conclude that M. nigrocinctus venom has a presynaptic action in the initial stages of intoxication followed by sub- and postsynaptic effects, the last being the most important cause of neuromuscular blockade. A direct action of the venom on muscle fibers may also contributes to the irreversible blockade. PMID- 10797873 TI - Capillary electrophoresis in mixed aqueous-organic solvents: effect of aprotic dipolar N,N-dimethylformamide and N,N-dimethylacetamide on acidity constants of substituted aromatic acids. AB - The change in the pKa values of substituted benzoic acids in binary mixtures of water with N,N-dimethylformamide (DMF) and N,N-dimethylacetamide (DMA), respectively, was determined up to a concentration of 80% (vol/vol) organic cosolvent. Such solvents are applied as media of the background electrolyte in capillary electrophoresis, with the intention of increasing the analyte solubility and the separation selectivity. The pKa values increase nonlinearly with increasing DMF and DMA concentration, and reach values between 2.5 and 3.7 pK units higher than in water at 80% organic solvent content. This change is discussed based on the concept of the medium effect, taking into account the particular stabilization of the species involved in the protolysis equilibria of the acids: HA, A-, and H+, in the different solvents. The contribution of the medium effect of the individual neutral acid HA on the delta pKa values is determined by solubility measurements. The increase in the delta pKa values, averaged for all five acids investigated, is compared in different mixed aqueous organic media, consisting of methanol, ethanol, 1-propanol, acetonitrile, DMF, and DMA, respectively. The comparison is based on the third-order polynomial fit of the delta pKa versus organic solvent concentration data. It is found that, in this respect, acetonitrile resembles the lower alcohols, and is dissimilar to the other dipolar, aprotic solvents, DMF and DMA. The similarity of the solvent systems is depicted by cluster analysis. PMID- 10797874 TI - Determination of short-chain free fatty acids in lipolyzed milk fat by capillary electrophoresis. AB - The objective of this study was to monitor the release of short-chain free fatty acids (FFA) from milk fat during hydrolysis with lipase using capillary electrophoresis. Sample and run buffer allowed FFA to be maintained in solution by using cyclodextrin and methanol. Indirect UV detection at 270 nm was used, employing p-anisate as a chromophore. Calibration curves constructed for each individual FFA followed linear relationships with highly significant (p < 0.01) correlation coefficients. Electrophoretic FFA profiles of fresh milk fat and lipolyzed milk fat showed marked qualitative and quantitative differences. Butanoic acid (C4) was found in a concentration of 64 ppm, while hexanoic (C6) and octanoic (C8) acids were found in concentrations of 3.8 ppm in fresh milk fat. After a 60-min hydrolysis with commercial lipase, FFA released from milk fat consisted mainly of high concentrations (ppm) of butanoic (C4) (900), followed by hexanoic (C6) (427), octanoic (C8) (282), decanoic (C10) (92), pentanoic (C5) (47), and dodecanoic (C12) (37.5) acids. Ratios of FFA that were associated with flavor balance were calculated. The application of CE for lipolysis monitoring in milk fat offers a simple and fast method for the determination of FFA. Quantitative data can be obtained in 20 min, including sample preparation. The lengthy and laborious steps required in traditional chromatographic techniques, such as lipid extraction, FFA isolation, and derivatization, were not required in this CE method. The implementation of CE for milk fat lipolysis monitoring may be a useful quality control tool for dairy flavor development and production. PMID- 10797875 TI - Applications of capillary electrochromatography analysis for formulated pesticide products. AB - The feasibility of using capillary electrochromatography (CEC) as a high efficiency reversed-phase separation technique has been demonstrated for the analysis of some pesticide formulation products. Some operating parameters of CEC analysis (organic modifier content, pH of the buffer, and sample diluent) were studied using commercially available capillaries packed with Hypersil (Phenomenex, Torrance, CA, U.S.A.) octadecylsilic (ODS) particles. It was found that the resolution decreases in linear fashion with the increase in percent acetonitrile in the sample diluent for neutral components if a combination of electrokinetic injection and pressure injection is used. Several practical applications of the CEC technique in the analysis of pesticide formulation products are described in detail. The results indicate that CEC, compared with HPLC, not only has higher efficiency, but is also practical, precise, and accurate in terms of simplicity, efficiency, recovery, and linearity. PMID- 10797876 TI - Determination of low-molecular-mass anions in coating samples by capillary electrophoresis with conductometric detection. AB - A method for the rapid analysis of low-molecular-mass anions in electrodeposition coatings has been developed, employing capillary electrophoresis with a nonsuppressed end-capillary conductivity detector. After optimization of the carrier electrolyte, quantitative determination of all analytes of interest (including inorganic anions and organic acids) was possible in less than 11 min. The results obtained by capillary electrophoresis did not show any significant differences with the manufacturer's data originating from the more time-consuming ion-chromatographic analysis. PMID- 10797877 TI - Capillary electrophoretic determination of carboxyhemoglobin concentrations in postmortem blood samples. AB - The use of capillary electrophoresis (CE) as an alternative to existing methods of quantitation of carbon monoxide (CO) in hemoglobin from postmortem blood samples is presented. The isolation of heme (the portion of the hemoglobin molecule in which CO binding takes place) from hemoglobin is described. Reduced (containing no gas molecules) heme and CO-heme isolated from hemoglobin standards were successfully separated using CE. Heme and CO-heme were also isolated from blood samples of accident victims and analyzed using CE. A quantifiable difference in the CO-heme signals from blood samples containing fatal and nonfatal levels of CO was observed. PMID- 10797878 TI - A chemometric approach to the detection of milk adulteration based on protein profiles determined by capillary electrophoresis. AB - The general objective of this study was to utilize chemometrics in the interpretation of capillary electrophoresis milk protein profiles, for the detection of pasteurized milk adulteration with rehydrated milk powder or a rehydrated dairy-based milk substitute. The specific objectives were 1) to collect quantitative data on major casein and whey proteins in authentic and adulterated milks in a single CE analysis; and 2) to apply a pattern recognition procedure, Soft Independent Modeling of Class Analogies (SIMCA), on collected CE protein data, for the development of a statistical model useful in the detection of pasteurized milk adulteration. Authentic samples were fresh milk collected from various farms over a period of six months. Adulterated samples were authentic fresh milk partially or totally substituted with rehydrated milk powder or a rehydrated commercial milk substitute at different levels. Quantitative protein data obtained by capillary free zone electrophoresis for beta lactoglobulin, alpha-lactalbumin, beta-casein, and alpha-casein of 86 samples, authentic and adulterated samples, were used as a training set to build a SIMCA multivariate statistical model. The detection of sample outliers was useful for the elimination of unusual samples and optimization of the multivariate model. From the 35 commercial pasteurized milks tested, which were treated as unknowns, a total of 14 samples (40%) were not assigned to the authentic or fresh milk group, meaning that these samples had some type of adulteration at the levels included in the training set (> 15%). Decision-making on detecting adulteration of unknown commercial pasteurized milk samples was eased since predictions were based on statistical probabilities. PMID- 10797879 TI - Determination of related impurities of bile acids in bulk drugs by cyclodextrin modified micellar electrokinetic chromatography. AB - A cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) method has been developed and validated for purity determination of two bile acids, ursodeoxycholic acid (UDCA) and deoxycholic acid (DCA). Quantitation of related impurities such as lithocholic acid (LCA), chenodeoxycholic acid (CDCA), cholic acid (CA), and DCA in UDCA and CA in DCA was performed. A running buffer containing 20 mM borate-phosphate, 50 mM sodium dodecyl sulfate (SDS), 2.0 mM beta-cyclodextrin, and acetonitrile was used. Modifiers were added to improve resolution and selectivity. The applied voltage was 25 kV and detection was performed at 185 nm. Validation parameters such as selectivity, linearity, repeatability, intermediate precision, limit of detection, limit of quantitation, and robustness were evaluated. The method was simple and proved to be useful for the purity testing of bile acids in bulk drugs. Good results were obtained for related impurities at concentration levels from 0.05 to 1.5% with respect to the main component, according to international requirements. PMID- 10797880 TI - Chiral separations of pharmaceuticals using capillary electrochromatography (CEC): an overview. AB - Enantiomers of bioactive molecules often differ in potency, toxicity, metabolism, and pharmacological actions. Capillary electrochromatography (CEC) is an emerging separation technique being investigated for use in chiral separations. CEC is a hybrid of HPLC and CE. CEC combines the electrophoretic mobility of CE with the partitioning mechanisms of HPLC. In this overview, several resolution mechanisms commonly used in CEC and the main parameters influencing the selectivity of the chiral separation will be discussed. Current applications of CEC in chiral separations of pharmaceuticals will be provided for each type of resolution mechanism. Finally, the advantages and limitations of CEC will be described, followed by the future outlook for CEC. PMID- 10797881 TI - Determination of drugs used as anti-Parkinson's disease drugs in urine and serum by capillary electrophoresis. AB - A new capillary electrophoresis method to determine simultaneously eight of the most important anti-Parkinson's disease compounds has been developed. The generic names of the drugs studied are benactyzine (BA), trihexyphenidyl (TP), fenpiverin (FP), diphemin (DF), scopolamine (BL), adiphenine (TS), diethylaminoethylester 1 phenylcyclopentane-1-carboxylate (EKK), and diethylaminoethylester tetramethoxydiphenylacetate (EKO). An untreated fused-silica capillary tube (75 microns i.d., 57 cm total length, 49.5 cm length to the detector) was used with detection at 190 nm. The optimal separation conditions were 50 mM phosphate buffer (pH 2.7) with 7 mM-beta-cyclodextrin, electrokinetic injection for 15 sec at 5 kV, temperature 25 degrees C, and 15-20 kV separation voltage. Complete separation of all compounds was achieved in less than 16 min. The procedure was applied for the determination in urine and serum. The limits of detection (LOD, S/N = 3) for serum were 209 (FP), 234 (EKO), 168 (DF), 182 (BA), 168 (TP), 220 (BL), 174 (TS), and 163 (EKK) ppb. The method can be used for the therapeutic drug monitoring of these central active cholinolytics in clinical laboratories. PMID- 10797882 TI - Profiling of cocaine by micellar electrokinetic chromatography. AB - The potential of micellar electrokinetic chromatography (MEKC) for the profiling of cocaine samples is described. An MEKC system containing sodium dodecyl sulfate (SDS) and methanol was optimized using a test mixture of cocaine, its common impurities (benzoylecgonine, norcocaine, tropacocaine, and trans cinnamoylcocaine), and several degradation products. The effect of pH, percentage modifier, and concentration surfactant on the separation has been investigated. The optimal separation buffer for cocaine samples consisted of 75 mM SDS, 17.5% methanol, and 25 mM borate (pH 8.3) and was well suited to separate components of diverse polarity in one run. Various cocaine seizures have been analyzed with the MEKC system and their signatures were compared. The electrokinetic chromatograms obtained were characteristic, and differences and similarities among the samples could easily be observed. Several impurities were identified in the samples by means of migration times and comparison of recorded and library UV spectra. The composition of the samples was determined semiquantitatively using relative corrected peak areas. PMID- 10797883 TI - Psychosocial and neuropsychological function in children with epilepsy. AB - This paper reviews the psychosocial and neuropsychological effects of epilepsy on children and families across environments in which children function, specifically home and school. Epilepsy is a chronic disorder, affecting one percent of the population, that alters neurocognitive functioning effecting learning, memory and family adaptation. A review of epilepsy and its impact on quality of life, family and school function and psychiatric comorbidity are discussed. PMID- 10797884 TI - The effect of hyperbaric oxygen treatment on postural stability and gait of a brain injured patient: single case study. AB - Hyperbaric oxygen (HBO) therapy has been found to reduce intracranial and cerebrospinal fluid pressures, and increase grey matter metabolic activity in patients with brain injuries. To date, few studies have quantitatively assessed the changes in the patient's functional outcomes following this expensive therapeutic intervention. The purpose of this case study was to examine the immediate and longer term changes in postural stability and gait in a 17 year old patient who sustained a traumatic brain injury, following administration of hyperbaric oxygen (HBO) therapy combined with physical and occupational therapy. The patient underwent assessments of postural stability and gait 1 week prior to HBO therapy, immediately following, and 6 weeks after completion of HBO therapy. Some improvements in postural stability were observed immediately following HBO, although these improvements were not evident 6 weeks later. Only slight improvements were noted in his walking abilities immediately following the intervention, with essentially little change evident 6 weeks later. The results of this do not support anecdotal evidence that there were substantial improvements in the subject's postural stability and gait following HBO therapy. PMID- 10797885 TI - The effect of language on maternal knowledge of the child's hearing status. AB - The aim of this study was to assess whether mothers whose first language is not English are as accurate as English speakers in reporting on their child's hearing. Some Health Trusts are using parental report following an educative process as part of their screening programme for hearing impairment. At present, in Wandsworth, London, UK, where there is a relatively high ethnic minority population, many of whom speak little or no English, the parental educative programme is carried out in English. There are many studies supporting the translation of the educative programme into appropriate minority languages but the authors wanted to assess whether doing so would actually improve the hearing impaired identification rate by the raised awareness programme. The presence or absence of concern in the two groups of mothers, one English speaking and the other non-English speaking, was elicited by direct questioning and its accuracy checked by formal hearing assessment. The results indicate that there is no significant difference in the reliability of maternal observation between the two groups in normal hearing and hearing impaired children. However, two-thirds of non-English speaking mothers of hearing impaired children were not concerned about their child's hearing status, compared to one-fifth of the English speaking mothers. Although the numbers of hearing impaired children are small, and the reason for the high maternal inaccuracy in non-English speakers in this group has not yet been ascertained, it seems reasonable to recommend translation of the parental educative leaflets in Wandsworth into minority languages. This recommendation is supported by other studies. PMID- 10797886 TI - Reduced body cell mass following severe head injury in children: implications for rehabilitation. AB - STATEMENT OF PURPOSE: Increased proteolysis, muscle catabolism and altered body composition have been well documented after severe head injury, but the extent of these effects in children, and whether they extend into rehabilitation, have not been studied. This study determined nutritional status and body composition, with particular reference to the body cell mass (BCM), of head injured children at entry into a rehabilitation programme, and compared body composition analysis with anthropometric nutritional assessment. METHODS: Nineteen head injured children (nine males, 10 females, mean age 9.1 +/- 4.3 years range 1.2-15.1 years) were measured for height, weight and total body potassium (TBK, a measure of body cell mass) on referral to rehabilitation after the acute phase (mean 38.1 days post-injury). Data was compared with expected normative data derived from healthy age and gender matched children. Nutritional status was determined by two separate criteria based on either anthropometric or body composition methods. RESULTS: The mean percentage of expected TBK for height was 84.4 +/- 15%, significantly below the clinically acceptable level for body cell mass (90% of expected). Using the anthropometric definition, only 1/19 was undernourished, whereas 12/19 had poor nutritional status using body composition (chi 2 = 7.58, p = 0.006). CONCLUSIONS: The data revealed a significant depletion in the metabolically active BCM in the presence of normal anthropometry, suggestive of significant muscle wasting. These findings have important pathophysiological and clinical implications in the rehabilitation of children following major head trauma. PMID- 10797887 TI - Functional impact of a rigid pelvic stabilizer on children with cerebral palsy who use wheelchairs: users' and caregivers' perceptions. AB - A within-subject ABA design was used to assess the functional impact of a novel wheelchair mounted rigid pelvic stabilizer (RPS) compared with a traditional wheelchair lap belt in a group of six children with cerebral palsy (mean age 10.4 years). The lap belt was worn during the two baseline phases which were each 3 weeks in duration. During a 5 week treatment phase the lap belt was replaced with the RPS device. Using the Canadian Occupational Performance Measure (COPM) each subject's self-evaluated performance ability and satisfaction with performance for five key tasks was measured at the end of each study phase. In addition, a structured weekly interview was conducted with the primary caregivers to assess perceived changes in their child's functional performance with each task. Results of repeated measures ANOVAs were statistically significant, indicating that the RPS as compared to the lap belt is a more effective device. The RPS allowed significantly better occupational performance and satisfaction with performance as measured by the COPM. Single-subject data analyses showed clinically significant changes in task performance and satisfaction with performance when the RPS was worn as compared to the lap belt for all subjects. Caregivers' perception of functional change ratings closely corresponded to subjects' self rated performance on specific tasks. Visual inspection of subject data also showed that, overall, the increased performance ratings for different tasks during the treatment phase decreased in the second baseline phase, when the RPS was removed, however, performance did not return to initial baseline levels. This suggests that the RPS has a facilitating effect for increasing physical functioning. These results are further discussed in terms of implications for practice, and future research. PMID- 10797888 TI - Cognition in 4-11 year old children in Turkey. AB - OBJECTIVES: The purpose of this study was to determine the relationship among age, sex and academic performance with cognitive ability on healthy age groups. METHODS: The subjects were 71 children, aged 4-11 years (x = 7.11). The children were divided into four age groups: 22 of the 71 children were at kindergarten and 49 children were in primary education. Thirty nine children were girls and 32 were boys. Twenty six children had good academic achievement and 23 did not. Each child was tested on the LOTCA battery by the Occupational Therapy Unit of Hacettepe University School of Physical Therapy and Rehabilitation. In this study, seven subtests were used from LOTCA (Loewenstein Occupational Therapy Cognitive Assessment). These are orientation for place, orientation for time, overlapping figures, praxis, reproduction of a two dimensional model, drawing a clock, and categorization. RESULTS: Statistical analysis of the relationships demographic variables, academic achievement and performance on the battery revealed that age was significant, sex was insignificant and academic achievement was significant when correlated with cognitive abilities. CONCLUSIONS: The results suggest that cognitive abilities tested show some differences. For this reason, in the school or other educational settings, children who have low academic achievement should be assessed also for cognitive abilities and then provided with occupational therapy. PMID- 10797889 TI - The assessment of Bruininks-Oseretsky test of motor proficiency in children. AB - OBJECTIVE: The primary objective is to research the relationship between motor abilities and demographic characteristics such as age and sex, in healthy children aged 4-11 years. METHODS: One hundred and twenty children in kindergarten (n = 30) and primary school (n = 90) were included in the study and evaluated by the Occupational Therapy Unit. All children were divided into four groups according to age, 4-5, 6-7, 8-9 and 10-11 years. The primary school children were classified according to academic learning, being successful or unsuccessful. In this study, Bruininks-Oseretsky test of motor proficiency was used to assess the gross motor skills and fine motor skills. These tests are running speed and agility (subtest 1/item 1), balance/walking forward heel-to-toe on walking line (subtest 2/item 6), bilateral coordination/tapping-foot and finger on same side synchronized (subtest 3/item 2), strength/standing broad jump (subtest 4/item 1), response speed (subtest 6/item 1), visual motor control/cutting out a circle with preferred hand (subtest 7/item 1), upper-limb speed and dexterity/pacing pennies in two boxes with both hands (subtest 8/item 2). RESULTS: When the children were classified according to sex, there were significant differences in subtests 6 and 7. According to academic learning, there were significant differences in subtests 2 and 8. When the results were evaluated due to age, important differences were found in subtests 1, 2, 4 and 8. CONCLUSION: It was seen that gross and fine motor skills in early childhood showed variety between age, sex and academic learning. The scores of motor abilities were better in successful children than unsuccessful children. The outcome of this study revealed that the Bruininks-Oseretsky test can be useful to investigate unexplored aspects of motor development. PMID- 10797890 TI - Orthopaedic rehabilitation in a case of Tay-syndrome. AB - This paper reports on the orthopaedic rehabilitation of a patient with Tay syndrome. Tay-syndrome is a rare monogen-inherited ektodermal dysplastic syndrome with ichtyosis, fragility of the hair and physical and mental retardation. The congenital ichtyosis is ubiquitous. Only the skin on the flexion side of the extremity joints are not involved (orthocerathosis combined with paraceratotic strings). In this case, a young boy developed bilateral subluxation of the hips and was not able to stand or walk freely. Contemporary pre- and neonatal care has prolonged the survival of newborns with severe genodermatoses, including this syndrome. In this case, it has provided the necessity for orthopaedic treatment of the problems caused by osteosclerosis and muscular spasticity. PMID- 10797892 TI - Neuromuscular control mechanisms and strategy in arm movements of attempted supranormal speed. AB - Rapid, goal-directed elbow flexion movements were examined under interacting conditions of inertial loading and resistance to movement initiation. The resistance ceased when movement began, resulting in quick release movements. Inertial load slowed the movement and lengthened the agonist and antagonist electromyographic (EMG) burst durations. The quick release resulted in larger accelerations but only minimal changes in peak velocity. Most aspects of the triphasic EMG pattern were little affected by the quick release, but the build up of agonist EMG and the corresponding rate of static force development differed markedly between load and quick release conditions. These and other data suggest that the specific pattern of agonist muscle activation is set according to neuromuscular constraints of the antagonist muscle and the expectation of movement dynamics. PMID- 10797893 TI - Intercepting moving objects during self-motion: effects of environmental changes. AB - The purpose of this study was to examine the role of background texture on an interception task during self-motion. Twenty-six participants modulated tricycle speed along one arm of a V-shaped track so as to intercept a ball approaching horizontally on the other arm of the V. Either a plain or a textured background (consisting of contrasting vertical stripes) was used. Velocity modulations occurred so as to keep the angle beta between the direction of heading and the line head-ball constant (constant bearing angle, or CBA strategy), indicating that this observer-environment relation might regulate the approach phase. In the textured condition, participants initially drove faster than predicted by the CBA model and compensated by slowing down in the second half. This is in line with the texture-induced overestimation of the ball velocity and implies that absolute velocity information is also used. PMID- 10797891 TI - Frances Anna Hellebrandt: physician, mentor, and pioneer in exercise physiology. PMID- 10797894 TI - The effect of submaximal exercise on recovery hemodynamics and thermoregulation in men and women. AB - Five women and 5 men were studied to examine the effects of submaximal exercise on thermoregulatory and hemodynamic variables during recovery in two environments: (a) control (C), 22 degrees C, 33% rh; and (b) hot humid (H), 32 degrees C. The participants exercised on a cycle ergometer at 60% of peak oxygen consumption for 35 min prior to 90 min of seated recovery. Sessions were identical, except for environment. Variables evaluated (p < .05) were: core temperature (TR), mean skin temperature (Ts), sweat rate (SR), heart rate (HR), stroke index (SI), cardiac index (CI), forearm blood flow (FBF), systolic blood pressure (SBP), and diastolic blood pressure (DBP). Men and women exhibited similar patterns of TR, Ts, and SR in both environments. Ts and SR (collapsed means for gender) were higher in the H than in the C. DBP was higher in men than in women throughout recovery in both environments. With combined means for gender, HR was higher in the H than in the C. CI, SI, FBF, and SBP were similar in both environments and returned to baseline within 15 min into recovery. These data suggest that heat dissipation during extended recovery was accomplished with similar contributions of cutaneous vasodilation and sweating in M and F. Furthermore, the moderate exercise level did not influence hemodynamics beyond 15 min of recovery in either environment. PMID- 10797895 TI - Tactical differences in problem representations and solutions in collegiate varsity and beginner female tennis players. AB - Youth tennis experts' tactical problem representations have been linked to their superior response selections during competition (McPherson & Thomas, 1989). To expand this research to adults, collegiate varsity (experts, n = 6) and beginner (novices, n = 6) female tennis players participated in an interview session to elicit problem representations and solutions to six tennis game situations. Interviews, transcribed verbatim, were coded for accuracy in solutions to each situation. Novices did not generate any solutions (actions), while experts generated similar and tactical solutions. Quantitative analysis of verbal data revealed expertise differences in problem representations and solution processes. Mann-Whitney U tests revealed that experts accessed more tactical and associated response selection and response execution concepts than novices. Findings indicate that experts will generate specialized cognitive processes to develop, monitor, and regulate their condition profiles and action plans during competition. PMID- 10797896 TI - The effects of ENDUROX on the physiological responses to stair-stepping exercise. PMID- 10797897 TI - Knowledge and attitudes related to diabetes and exercise guidelines among selected diabetic children, their parents, and physical education teachers. PMID- 10797898 TI - Characteristics of Maxiflex Model B springboards revisited. PMID- 10797899 TI - Comparison of the criterion-related validity of sit-and-reach tests with and without limb length adjustment in Asian adults. PMID- 10797900 TI - [Advantages of neuroendoscopy in the management of non-communicating hydrocephalus]. AB - INTRODUCTION: Non-communicating hydrocephalus is defined as a blockade of the physiological cerebrospinal fluid (CSF) flow from the choroid plexus to the subarachnoid space. Usually, a ventricle peritoneal shunt with a valve is employed as treatment but this does not provide a physiological restoration on the CSF flow. OBJECTIVE: The aim of the study is to introduce a more physiological solution by means of endoscopic technique. We also pretend to find out some prognostical factors related to successful treatment. PATIENTS AND METHODS: We present a prospective non-randomized series of 32 endoscopic III ventriculostomies performed in patients with non-communicating hydrocephalus. When the blockade was due to a tumor, an endoscopic biopsy was performed at the same surgical time. In the same way, if the cause was a ventricular hematoma, an endoscopic aspiration was performed. The patients follow-up was two years. RESULTS: The procedure was considered successful (83.8% of the cases) when the clinical symptoms resolved without ventricle-peritoneal shunt. We study the relationship of the blockade level and the highest value of intracranial pressure during endoscopy with the results of the procedure in terms of success and morbidity. We have found significant relation between high pressure during endoscopy and the practice of a complementary biopsy or hematoma evacuation. CONCLUSIONS: Endoscopic third ventriculostomy provides an effective and physiological solution to non communicating hydrocephalus. Complementary endoscopic procedures can be associated at the same surgical time, however this association can be related to increased values of intracranial pressure during surgical performance. PMID- 10797901 TI - [Auditory brain stem potentials evoked by electrical stimulation of cochlear nuclei during central auditive implant setting]. AB - INTRODUCTION: The implantation of a stimulator over the surface of the cochlear nuclei can provide a partial auditive capability to patients deaf due to a bilateral lesion of the 8th nerve. Intraoperative monitoring of short latency electrically-evoked potentials is useful to asses the correct positioning of the implant, specially when there are anatomical distortions. PATIENTS AND METHODS: Evoked potentials from two patients with type II neurofibromatosis were recorded during implantation surgery, using a blanker system to eliminate the stimulus artifact. RESULTS: In both cases, two-peak vertex-positive responses were obtained, with latencies between 0.4-0.5 and 1.2-1.4, respectively. This responses are similar to the most frequent response described. DISCUSSION: Intraoperative monitoring of electrically-evoked auditory brainstem responses is a useful technique to confirm the correct positioning of the cochlear stimulator. The study of the morphology of these responses can help to understand the mechanisms involved in the generation by the brainstem of the auditive evoked potentials. More experience on this subject is needed to establish a correlation between intraoperative results and postoperative stimulator function. PMID- 10797902 TI - [Long-term follow-up of childhood epilepsy treated with vigabatrin outside of clinical trials]. AB - INTRODUCTION: Experimental conditions are not mirrored by clinical practice. OBJECTIVE: To study the efficacy of vigabatrin in the usual conditions of everyday clinical practice. PATIENTS AND METHODS: Retrospective review of all epileptic patients treated with vigabatrin in a neuropediatric outpatient clinic. OUTCOME MEASURES: a) Persistent seizure frequency reduction > or = 50%, and total control of seizures. Potential predictors of response were studied by logistic regression. b) Duration of VGB therapy, studied by the Kaplan-Meier method and its associated log-rank test. RESULTS: 113 patients with: partial symptomatic epilepsy (38%), partial cryptogenic (25.6%), partial idiopathic (6%), West syndrome (14%), Lennox-Gastaut syndrome (6%), other syndromes (9.7%). Reduction of seizure frequency was attained by 60% of patients at 3 months, sustained during 12 months by 40%, and during 5 years by 14%. Total control of seizures was present in 33% of patients at 3 months, persisted 12 months in 18% and 5 years in 2%. Independent predictors of a poor outcome were generalized seizures (except infantile spasms) and cerebral palsy, among others. The probability of continuing vigabatrin (VGB) therapy was 78% at 6 months, 55% at 2 years and 32% at 5 years. Duration of therapy was modified by early therapeutic response and antecedent of status epilepticus, among others. Adverse events were recorded in 18.5%. Visual fields were not studied in these series. CONCLUSIONS: For the time being, VGB treated patients belong to the difficult-to-treat group. Percentages of responders depend upon duration of follow-up. PMID- 10797903 TI - [The development of the human cerebral cortex. A cytoarchitectonic theory]. AB - INTRODUCTION: In order to understand the over-all organization of the neocortex, its development in the embryos of certain mammals has been studied using the Golgi method. DEVELOPMENT: The neocortex starts its development with a primary plexiform layer in the telencephalon, which precedes and is essential for formation of the cortical plaque. Layer I and the sublayer are derived from this primary plexiform layer which represents the primitive cortical organization shared with reptiles and amphibians. The other layers (II, III, IV, V and VI) are derived from the cortical plaque which is an innovation occurring in mammals. During the development of the cortical plaque, migration, early differentiation and morphological and functional maturity of the neurones occur. The neurones, guided by the radial glia, reach layer I, develop an apical dendrite and establish contact with the cells of Cajal-Retzius, after which the migratory neurones assume a common pyramidal morphology. During ascending cortical maturity, controlled by the thalamus, the neurones acquire their specific morphology and function. The cortical plaque represents a biologically non specific stratified nucleus which increases the number of pyramidal layers during the evolution of the mammal. CONCLUSIONS: In this paper we emphasis the importance of the Cajal-Retzius cells in neuronal migration, formation of the cortical plaque, morphology of the pyramidal cell and ascending stratification- morphological and functional--of the neocortex. We also introduce a new cytoarchitectonic theory of the phylogenetic evolution of the mammalian neocortex. PMID- 10797905 TI - [Genetic encephalopathies in a newborn]. AB - The clinical manifestations of neonatal encephalopathies lack specificity and may present with predominantly seizures, hypotonia or coma. We have selected several examples of neonatal encephalopathies which are of special interest because of their genetic origin and present their clinical features, typical course and, when available, treatment. PMID- 10797904 TI - [Congenital vascular malformations in childhood]. AB - The intracranial congenital vascular pathology, arterial, arteriovenous and venous, is reviewed, including the cavernomas as well. It is commented the possible origin, the clinical, histological and neuroradiological alterations as well as the possible treatment, when necessary, of every entity. The contribution of the magnetic resonance (MR), especially the angiographic MR (AMR) has been conclusive to the diagnosis. There have been considered six main groups which include the main entities: 1. Arterial anomalies; 2. Arteriovenous malformations; 3. Malformations of the Galen vein; 4. Venous malformations, and 5. Intracranial cavernomas. PMID- 10797906 TI - [Cerebral hemorrhage in full term newborns]. AB - INTRODUCTION: Cerebral hemorrhages in full-term newborn babies are an important factor in neonatal morbidity and mortality and very frequent in premature babies. In full-term newborn the frequency is reduced to 1-2% and the aetiopathogenesis is basically related to birth trauma. OBJECTIVE: To identify the clinical forms of cerebral hemorrhages in full-term newborn before taking prophyllactic and/or therapeutic measures if possible. DEVELOPMENT: Based on the integrated physio pathological model of Wigglesworth and Pape, two anatomo-pathological patterns were established according to gestational age. Topographic classification was done in full-term newborn according to site (subarachoid, subdural, intraventricular, cerebellar and intraparenchymatous). We studied the pathogenesis, clinical features and diagnosis of each of these sites and emphasize the importance of neuroimaging. CONCLUSIONS: The diagnostic approach proposed permits an aetiopathogenic and therapeutic view which currently permits improved prognosis and even cure in many cases. PMID- 10797907 TI - [Complications of intracranial hemorrhage in full term newborns]. AB - The incidence of various types of intracranial hemorrhage is different in full term infants than those that occur in small premature infants. Consequently, the complications are for the most part different. The previous article described the types of hemorrhage occurring at term, as well as their signs and symptoms. This brief review will focus on the complications that may arise from these. Examples are given of superficially occurring hemorrhages and of those that arise deeper in the parenchyma of the brain. The actual bleed is the acute event that may become subacutely complicated by virtue of its position or by extension. This occurs as part of the natural process or as a result of injudicious intervention. There follow chronic complications: cerebral palsy, intellectual deficit and seizure disorder which remain for the patient's lifetime. PMID- 10797908 TI - [Treatment and prevention of cerebral hemorrhage in newborns]. AB - INTRODUCTION: The risk factors leading to the occurrence of intracranial hemorrhages in premature newborn babies are well known. However, in full-term babies this has not been so systematically studied. Although the incidence is lower it is not rare and they are more varied in the form of presentation than in premature babies. Thus, as well as the classical intraperiventricular hemorrhages, subarachnoid and intraparenchymatous hemorrhages also occur relatively often, particularly the subarachnoid kind. The aetiopathogenesis of the hemorrhages is different, so preventive measures are also different, based particularly on avoidance of perinatal asphyxia and traumatic manoevres during labour. DEVELOPMENT: A study by our Department, in which risk factors associated with 17 newborn babies of over 35 weeks gestation who had a history of intracranial hemorrhage in the early neonatal period showed that over half the cases (58%) had a history of acute foetal distress. A quarter had congenital cardiopathy (detected at birth) and the same number had a respiratory distress syndrome due to various causes, particularly pneumothorax and infections. Regarding treatment of the acute phase, it is essential to maintain cerebral perfusion and monitor both arterial hypertension and hypotension, control the intracranial pressure (avoiding increases) and avoid fluctuations of cerebral blood flow. CONCLUSION: Follow-up by means of serial echographs makes early detection of dilatation and hydrocephaly possible and permits suitable suitable treatment rapidly and with immediate monitoring of its efficacy. PMID- 10797909 TI - [Epidural abscess of the cervical spine secondary to dental abscess]. PMID- 10797910 TI - [Gliomatosis cerebri. MRI, SPECT and the study of pathology]. PMID- 10797911 TI - [Leber's hereditary optic neuropathy and its possible relation to a recent epidemic in Cuba]. AB - INTRODUCTION: This century, the greatest epidemic affecting the nervous system was notified in Cuba seven years ago. At the present time the epidemic continues although to a lesser extent. The clinical findings of the illness were mainly bilateral optic neuropathy sometimes accompanied by other symptoms and peripheral neuropathy. The similarity of the optic form of the illness with Leber's hereditary optic neuropathy, and their common risk factors, were obvious from the beginning. PATIENTS AND METHODS: Statistics from the national reference department of neuro-ophthalmology of the Instituto de Neurologia de Cuba were reviewed. From these it was evident that the number of cases of Leber's hereditary optic neuropathy had increased in recent years, coinciding with the period of epidemic and endemic Cuban neuropathy. Many of these patients had previously been diagnosed as having epidemic optic neuropathy. RESULTS: We describe the characteristics of a group of these patients and discuss the differences and possible relationship between the two conditions. In the case of epidemic optic neuropathy, there is strikingly simultaneous loss of vision, less visual changes with much smaller cecocentral scotomas, loss of ganglion fibres of the retina around the papillomacular bundle, a good response to multivitamin treatment, and increased frequency of association with peripheral sensory neuropathy. CONCLUSION: This analysis reinforces the hypothesis that many patients with Leber's hereditary optic neuropathy, which started at the time of the epidemic, were incorrectly classified as suffering from this, and also perhaps their condition worsened due to the toxic nutritional features common to both conditions. PMID- 10797912 TI - [Venezuelan equine encephalitis. 1995 outbreak: clinical profile of the case with neurologic involvement]. AB - INTRODUCTION: Venezuelan equine encephalitis virus has caused periodic epidemics and epizootics in the American continent since the 1920s. Such events have been profusely documented from the epidemiologic point of view, however, reports concerning the clinical features of this disease are rather scarce. OBJECTIVE: To analyze the clinical characteristics evidenced by Venezuelan equine encephalitis patients from Zulia state (western Venezuela) studied during the outbreak that occurred in Colombia and Venezuela in 1995. These cases, classified as complicated, were hospitalized at the Hospital Universitario de Maracaibo, state of Zulia, Venezuela. PATIENTS AND METHODS: The clinical charts of 313 Venezuelan equine encephalitis patients hospitalized during the period January 1st 1995 March 31st 1996 were reviewed. These cases accounted for 2.82% of 11,072 patients that were medically assisted during the outbreak. The following variables were analyzed: age, gender, signs and symptoms, contact history, complications and evolution. RESULTS: Intracranial hypertension signs became eloquent in 55.9% of these patients. Neurologic complications were represented by two cases of cerebellitis, two cases of meningoencephalitis and one case of encephalomyelitis. The mortality rate was 1.7%. CONCLUSION: Our results corroborate the benign evolutionary profile that is typical of this entity. PMID- 10797914 TI - [Evaluation of children with developmental disorders by evoked potentials and event-related potentials]. AB - INTRODUCTION AND DEVELOPMENT: Evoked potentials evaluation of children with central nervous system developmental disorders o dysfunctions is better than clinical and imaging assessment for determination of: 1) conceptional age, 2) auditory and visual threshold, 3) integrity of sensory pathways, 4) neurologic motor outcome, and 5) course and evaluation of treatment of neurometabolic disorders. CONCLUSIONS: However, it is not the same for predicting or making the diagnosis of the majority of primary developmental disorders (autism, receptive and expressive language disorders, learning disability, attention deficit disorders and Gilles de la Tourette syndrome. PMID- 10797913 TI - [Gamma-sarcoglycanopathy:two new cases in a gypsy family family in Spain]. AB - INTRODUCTION: Included under the heading of limb girdle muscular dystrophy is a heterogeneous group of myopathies which share the same phenotype characteristics. The illness is of early onset, progressive and basically involves muscles of the shoulder and pelvic girdles. Recent identification of muscle proteins and the genes which codify them has led to new classification of these conditions according to their genetic characteristics. It is currently accepted that there are two major groups: the dominant and recessive forms. The latter includes type 2C limb girdle muscular dystrophy associated with chromosome 13, where the gene for gamma-sarcoglycan is found. This protein belongs to the glycoprotein complex associated with dystrophin. Recently a new mutation has been identified, the C283Y, exclusive to the Gypsy race, which affects this gene and therefore the alterations in gamma-sarcoglycan produced by it. CLINICAL CASE: We describe two patients, Gypsy brothers, who complained of myopathy, which they had had for some years, compatible with this condition and in whom the C283Y mutation had recently been detected. CONCLUSIONS: We describe another Spanish Gypsy family, all members of which have the characteristic mutation. We emphasize the importance of genetic studies in all cases of myopathy which have not been fully diagnosed. PMID- 10797915 TI - [Prognosis of bacterial meningitis]. AB - OBJECTIVES: To review the literature on the risk factors for bacterial meningitis (BM) and evaluate these factors in children diagnosed as having BM in the Paediatric Department of the University Hospital of Maracaibo between 1996 and 1998. PATIENTS AND METHODS: We made a retrospective study of children diagnosed as having BM in the University of Maracaibo. We evaluated different factors related to BM. RESULTS: From 1 January 1996 to 31 December 1998 a total of 152 children were diagnosed as having bacterial meningitis; 69.7% were boys and 30.3% were girls. The commonest causal germ was Haemophilus influenzae. Disorders of sensation (42.7%) and signs of meningism (32.8%) were the most frequent neurological alterations. The main laboratory findings were leucocytosis (51.3%), thrombocytosis (49.3%), pleocytosis (70.3%), cerebrospinal fluid protein (49.3%) and low cerebrospinal fluid glucose (72.8%). Twelve patients (7.5%) died. Of this group, 10 were under one year of age and had septic shock. CONCLUSIONS: BM represents a major group of hospital admissions in everyday paediatric practice. Although the mortality has decreased, an increased risk of sequelas must be borne in mind. Clinico-neurological examination on admission and simultaneous analysis of laboratory investigations allows identification of prognostic indicators of morbidity and mortality. PMID- 10797916 TI - [Neurological involvement in free living amebiasis]. AB - INTRODUCTION: Free-living amoeba infection is today a known and outgrowth disease, which is suspected when exists a special skin lesion. DEVELOPMENT: Balamuthia mandrillaris is one of these amoebas recently categorized and identified as a cause of meningoecephalitis granulomatous and chronic. Human infection: until 1997 in Cayetano Heredia Hospital (Lima, Peru) was described 24 cases of Balamuthia mandrillaris infection, 10 under 15 years old. All had skin lesion and all died, 16 had conscious impairment, 8 intracranial hypertension and 8 seizures. CONCLUSIONS: The free-living amoebas disease is now an outgrowth illness that depends on its worldwide distribution and association with immunocompromised host. The Balamuthia mandrillaris infection is a new form responsible for a fatal disease and clinical suspected is possible by the occurrence of skin lesion. Is very important to promove in Latin America collaborative studies in order to future actions. PMID- 10797917 TI - [Aids neurologic manifestations in childhood]. AB - INTRODUCTION: Children with AIDS get infected mainly by vertical transmission. DEVELOPMENT: That was what happened in 90% of the cases in a series of 340 HIV+ children followed at Hospital de Clinicas de Porto Alegre, Brazil. Currently, after the use of prophylactic treatment during pregnancy and the six first weeks of life, our transmission rate is 3% to 4%. The incidence of neurologic complications in the vertical transmission group was 49% in our prospective series, and due either to immunosuppression or primary neurological disease. Neurologic changes secondary to HIV constitute a complex syndrome, manifested by various degrees of cognitive, motor and behavioral delay. Encephalopathy can be either progressive or static. In our series, encephalopathy occurred in 32.5% of cases, developmental delay in 42.5%, seizures in 6%, speech delay in 5%, headaches in 2.5% and behavioral disorders in 2%. Central nervous system infections happened in 33.8% of our cases: acute bacterial infections in 11%, cytomegalovirus in 6.8%, toxoplasmosis in 5.9%, cryptococcal in 5%, tuberculous in 3.8% and syphilis in 1.3%. Hemorrhagic cerebrovascular accidents were seen in 2.5% of our cases, and peripheral neuropathy in 5%. Seventy percent of our cases had abnormalities in the CSF, 75% in brain CT scans and 45% in EEGs. CONCLUSIONS: Treatment of children with AIDS and neurologic manifestations was based on specific drugs aimed at controlling viral replication. Best response was obtained with combined use of AZT and other anti-retroviral neuro-protective. PMID- 10797918 TI - [Neuroanatomical, neuroradiological and functional magnetic resonance imaging correlates of developmental dyslexia]. AB - Neuropathological data in dyslexia has demonstrated alterations in the symmetry normally present in the planum temporale, as well as microdysgenesis in superficial cortical layers and disruption of the cytoarchitecture in subcortical structures. Neuroradiological and functional neuroimaging studies in dyslexia are consistent with localization of dysfunction to the temporoparietal junction. Present neuropathological and neuroimaging data support the concept that the fundamental problem in developmental dyslexia is a phonologic deficit. Present neuroimaging technology has increased our understanding of the pathophysiology of dyslexia. Functional neuroimaging may allow for the investigation of the effects that treatment of dyslexia has on brain plasticity. PMID- 10797919 TI - [Neurological assessment for learning disability]. AB - Learning disabilities present a diagnostic and therapeutic management challenge that requires an interdisciplinary approach initiated and coordinated by the child neurologist. An optimal evaluation for these conditions must include an extensive knowledge of the types and range of these disorders, a complete neurological and laboratory examination, a neuropsychological assessment tailored to the individual needs of the patient, a speech pathological evaluation when pertinent, and special attention to the frequent co-occurrence of more than one condition susceptible of interfering with learning. PMID- 10797920 TI - [Epidemiology of neurocysticercosis]. AB - INTRODUCTION: Cysticercosis remain an important health problem in developing countries. Its transmission is related to soil contamination with human feces. This parasitosis is found in Africa, Asia and Latin America, where the greatest incidences are seen in Mexico and Brazil. DEVELOPMENT: Human cysticercosis is acquired from the ingestion of ova of Taenia solium, excreted by human carriers in their feces, followed by the development of cyst in human tissue. The risk of contamination with Taenia ova is related to the contact with Taenia solium carriers. Recently, it has been shown that, in humans, the most common route of infection is ingestion of Taenia solium eggs from contaminated food or water. In United States of America and Europe, the frequency of cysticercosis is increasing due to increasing immigration and more frequent travels to endemic regions. The infected individual becomes a carrier and source of infection by oral-fecal contamination. CONCLUSIONS: Prevention is the single most important factor in reducing the frequency of cysticercosis. The transmission cycle of cysticercosis could be interrupted by improving sanitary conditions, and eliminating human cysticercosis. The treatment of Taenia carriers could be effective in prevention of cysticercosis, by reducing the excretion of its eggs and, so, reducing the risk of infection. At long date, education--washing hands before eating and after using bathroom, drinking boiled water--and improvements in sanitation, health care and socio-economic status are essential in prevention of human cysticercosis. PMID- 10797921 TI - [Clinical findings in active cerebral neurocysticercosis in pediatrics]. AB - OBJECTIVE: We describe our experience in the study of active cerebral cysticercosis in paediatrics over a period of 6 years, involving a total of 85 cases, which were studied from the clinical, immunological and radiological points of view. The aim of this communication is to present the clinical characteristics of the disorder and its course when symptomatic treatment is given. DEVELOPMENT: The commonest age of presentation was school age. However, we had a large number of children under 4 years old. This shows that there is considerable endemicity in a group in which the disorder is considered to be less frequent. There were slightly more females than males, which is in accordance with another Mexican series. The predominant socio-economic level was poor (59%). The patients come from urban areas more and more frequently. The commonest signs were: convulsive crises, intracranial hypertension and headache without intracranial hypertension. The commonest type of crisis was partial, in its different forms. CONCLUSIONS: The course was mild and satisfactory in 90% of the patients. The good clinical progress was independent of the use of antiparasitic treatment. PMID- 10797922 TI - [Neurocysticercosis in pediatrics. Prognosis, treatment and prevention]. AB - INTRODUCTION: Neurocysticercosis is an endemic disease caused by the larva of Taenia solium, which accidentally finds its way into the central nervous system of human beings. It is not found in any zone in particular and this leads to clinical pleomorphism. This makes treatment in paediatric patients even more difficult than in adults, because of their growth and development. DEVELOPMENT: There is controversy over the use of cestocide drugs, which should not be used in general except in specific cases such as cysticercosic meningitis and when there is intracranial hypertension. In other cases symptomatic treatment is recommended. In few cases is surgery worthwhile. Prognosis is good in most patients. CONCLUSION: Prevention by hygienic-dietetic measures is the basis for control and eradication. PMID- 10797923 TI - [Syndrome of Ohtahara]. AB - INTRODUCTION: A literature review using the term Ohtahara syndrome and early infantile epileptic encephalopathy revealed 51 cases. DEVELOPMENT: The characteristics of these cases were: 1) early onset of seizures; 2) tonic seizures; 3) poor response to treatment; 4) mental retardation; 5) poor prognosis; 6) burst-suppression EEG pattern; 7) evolution to West syndrome, and 8) multiple causes. These characteristics are non specific. No cause was found in most cases. The onset of seizures was between 7 hours and 86 days of age. The most frequent brain imaging abnormality was diffuse atrophy. ACTH was effective in a few cases and hemispherectomy was successful in one case. CONCLUSION: We believe that Ohtahara syndrome and early myoclonic encephalopathy are the same entity. PMID- 10797924 TI - [West syndrome]. AB - The author presents a review and actualization of West syndrome related knowledges, ethiological issues, clinics, and the EEG tracings. It is also include a literature review about different therapeutic treatments emphasizing the use of vigabatrin. PMID- 10797925 TI - [Lennox-Gastaut syndrome]. AB - INTRODUCTION: The Lennox-Gestaut syndrome is classified as an epileptic syndrome shown by the presence of various types of generalized seizures (tonic, atonic and atypical absences) which appear at a certain age (1-8 years), with an interictal EEG showing an abnormally slow basic rhythm interrupted by slow spike-and-wave complexes (< 3 Hz) and progressive mental deterioration. DEVELOPMENT: From the aetiological point of view there are cryptogenic (25%) and symptomatic (75%) forms. There is a previous history of West syndrome in 9.4-30% of the symptomatic cases. The commonest types of seizures are tonic (17-95%), atypical absences (17 60%) and atonic (25-56%). The mixed form of an epileptic state with typical absences and tonic seizures is the most frequent (27%). Follow-up studies show that in 90% and 100% of cryptogenic and symptomatic patients, respectively, mental retardation develops and the initial seizures persist in 67% and 45% of the patients with cryptogenic and symptomatic forms respectively, when they become adults. CONCLUSIONS: There is still no successful treatment for these seizures and progressive mental deterioration occurs even when using the newer anti-epileptic drugs. Electrical stimulation of the vagus nerve seems a promising possibility but further experience is necessary. PMID- 10797926 TI - [Urgent neurological consultation in children with systemic cancer]. AB - Neurologic complications are common in children with cancer but the literature dealing with this subject is sparse. Utilizing a symptoms and signs approach we review the most common causes for requesting an urgent neurologic evaluation for this population. The spectrum of neurologic symptoms in children with cancer differs from adults and requires the consulting neurologist to have a thorough knowledge of childhood cancer and its effects on the nervous system. PMID- 10797927 TI - [Status epilepticus in children. Experience with intravenous valproate. Update of treatment guidelines]. AB - INTRODUCTION: The treatment in the acute phase of seizures and status epilepticus is a neurological emergency situation. Aggressive treatment and resuscitation maneuvers against the seizure may be required to avoid progression towards status epilepticus, with the high risk for the patients outcome this involves. DEVELOPMENT: The use of rectal diazepam has supposed a great advance in the management of acute seizures. Intravenous sodium valproate has become available in these last years to control seizures and status epilepticus. Given at 20 mg/kg as single dose and with maintenance rate of 1 mg/kg/hour by intravenous infusion in 19 paediatric patients (1 day-7 years) status epilepticus was controlled in 58%, with reduction of status in 26%. Tolerance was good and there was no evidence of important adverse effects. CONCLUSION: In view of our experience, we present a new protocol for the treatment of seizures and status epilepticus in childhood. PMID- 10797928 TI - [Anterior ischemic optic neuropathy in children]. AB - A 5-year old girl with history of chronic renal failure due to dysplastic and polycystic kidney disease treated with peritoneal dialysis since birth wake up the the day after an scheduled uncomplicated peritoneal dialysis except for asymptomatic arterial hypotension with bilateral blindness. Neurological examination was normal except for absent light perception and tracking of optokinetic tape, dilated (7 mm) and unresponsive to light pupils, pale and slighted elevated disks with blurred margins and pale retina with irregular abnormal appearing vessels. Spinal fluid examination revealed no abnormalities except for slight pleocytosis (12 WBC, 68% mononuclears and 32% lymphocytes). MRI with gadollinium and MRA were normal. She received intravenous methyprednisolone for 3 days and intravenous and intraperitoneal antibiotics and intravenous acyclovir without improvement. She also received topical dorsolamide hydrochloride. Two year later she remained blind without light perception. Final diagnosis was anterior ischemic optic neuropathy. Pathogenesis, clinical picture, differential diagnosis and treatment of this condition in children are discussed. PMID- 10797929 TI - [Opercular epileptic syndrome: an unusual form of benign partial epilepsy in childhood]. AB - INTRODUCTION: Functional opercular syndrome in childhood is an exceptional form of presentation of benign partial epilepsy with centro-temporal rolandic spikes (BECRS). CLINICAL CASES: We studied the evolution of four patients, three of them followed for more than 15 years. Two were siblings, and their father suffered from BECRS with permanent language problems (verbal dyspraxia) and difficulty of protunding his tongue in adulthood. A third patient suffered benign familial neonatal convulsions (BFNC). In all four patients the actual illness begun as a BECRS with opercular troubles as an ictal phenomena. At about four years of age, the opercular disfunction became evident, with severe drooling, facial hypomobility and speech disturbance which waxed and vanished along weeks, months or years, apparently not ictal. Antiepileptic drugs not only were unable to control this situation but also, some of them, like carbamazepine, even worsened the opercular disfunction, increased the number of seizures and enhanced the neuropsychologic disfunction. Only clobazam could achieve the control on opercular disfunction. After 16 years, no further treatment was needed for all patients. There were some permanent sequelae, as speech and orolingual dyspraxia and different neuropsychologic problems. CONCLUSION: Of noteworth the best performance was attained by the patient treated with clobazam on monotherapy. PMID- 10797930 TI - [Landau-Kleffner sydnrome]. AB - The Landau-Kleffner syndrome consists of the association of paroxysmal EEG abnormalities, more marked during sleep, acquired aphasia usually of receptive type, and epileptic seizures in three quarters of cases. Additional features include behavioral disturbances that may even present with autistic features, cognitive regression of variable degree, and sometimes motor difficulties, indicating the pervasive nature of the disorder. The epileptic activity, as manifested by the EEG, seems responsible for the pervasive dysfunction. Treatment with antiepileptic drugs is often ineffective. ACTH and corticosteroids are currently regarded as the best therapy but surgical treatment by subpial transection is being actively studied. PMID- 10797931 TI - [Epilepsy treatment by vagal stimulation]. AB - INTRODUCTION: Vagal nerve stimulation is the latest therapeutic modality for the treatment of epilepsy. It consists of a lead implanted in the left vagal nerve which is connected to a subcutaneous stimulator implanted in the left axillary or pectorial region. DEVELOPMENT: The stimulator is programmed to intermittently stimulate the vagal nerve throughout the day and a magnet also allows the patient to control the stimulation from the outside. This treatment has been used in patients with intractable partial seizures who are not candidates for epilepsy surgery. The results reported have varied but in general the procedure appears promising with at least 50% of the implanted having over 50% improvement in their seizure frequency and many having complete control without significant side effects. CONCLUSION: Further review of the results are still needed to fully determine the true value of this treatment and to identify the subgroups of patients which will benefit the most. PMID- 10797932 TI - [Hydranencephaly. A case report]. PMID- 10797933 TI - [Antiepileptic effect of Plectranthus amboinicus (lour.) Spreng. (French marjoram)]. PMID- 10797934 TI - [On the relationship between Chiari malformations and hydrosyringomyelia]. PMID- 10797935 TI - [Unilateral carotid sinus hypersensitivity as a sign of occlusion of the internal carotid artery]. PMID- 10797936 TI - [Bulbar hematoma]. PMID- 10797937 TI - [Ischemic stroke as first manifestation of chronic myelogenous leukemia]. PMID- 10797938 TI - From endometrial receptivity to infertility. AB - Implantation of the blastocyst in endometrium requires establishment of a coordinated molecular dialogue between the embryo and the endometrium. Factors instrumental in the preparation of a receptive endometrium are derived from the hypothalamic-pituitary-gonadal axis. These factors modulate the expression of genes that drive the endometrium throughout the characteristic menstrual cycles. During each menstrual cycle, a series of coordinated, architectural, morphological, cytochemical, and molecular changes ultimately lead to the preparation of a receptive endometrium during the putative "receptive period" or "implantation window." It is during this critical period that a proper dialogue can be established between an intrusive blastocyst and a receptive endometrium. If, for any reason, this dialogue is not established or is perturbed, the embryo is aborted. The natural fate of the receptive endometrium, in the absence of implantation, is development of a second set of changes that ultimately lead to menstruation. The identity of the molecular repertoire that makes endometrium receptive to implantation and/or lead to menstruation is being revealed and broadly includes cytokines, heat shock factors, adhesion molecules and matrix metalloproteases. We identified a novel gene of the transforming growth factor beta, superfamily of molecules, the so-called endometrial bleeding--associated factor or ebaf, whose expression is confined to the late secretory and menstrual phases. Various forms of female infertility were associated with dysregulated expression of ebaf during the implantation window. The findings show an occult molecular defect of endometrial receptivity that seems to be due to dysregulated and premature expression of a member of the premenstrual molecular repertoire. The dysregulated expression of ebaf may assist in the identification, prognostication, and monitoring of treatment of infertile women. PMID- 10797939 TI - Genetic control of uterine receptivity during implantation. AB - Implantation involves complex molecular interactions between implanting blastocysts and the hormonally primed uterus. Gene targeting allows the generation of mice lacking a specific gene or genes and has proved to be of considerable value when combined with classical physiology in understanding many biological questions, such as the process of implantation. In this article, we review genes that have been demonstrated by gene targeting in mice to be required in the uterus for implantation. In particular, we focus on a specific class of developmental control genes, the mammalian Hox genes, and their role in this process. Lastly, we attempt to synthesize current knowledge about the genetic control of implantation and to build a working genetic model for the implantation pathway. PMID- 10797940 TI - Dynamics of uterine glycoconjugate expression and function. AB - Embryo attachment to the apical surface of the uterine epithelium is an event found in all mammalian species. Consequently, aspects of this process may be shared and considered as general principles in implantation strategies across species. This review focuses on studies implicating mucin glycoproteins disposed at the apical surface of uterine epithelia as antiadhesive molecules that block embryo attachment. As such, mucins must be removed, at least locally, to permit intimate contact between trophectoderm and uterine epithelia. Subsequently, we consider the role that heparan sulfate proteoglycans (HSPGs) and HSPG-binding proteins play in tethering embryos to the apical surface of uterine epithelia during the attachment process. PMID- 10797941 TI - Trophinin, tastin, and bystin: a complex mediating unique attachment between trophoblastic and endometrial epithelial cells at their respective apical cell membranes. AB - Embryo implantation is a complex process consisting of multiple cross-talks between maternal and embryonic cells. Defining the mechanisms underlying implantation at molecular level is challenging task in reproductive biology. In order to identify molecules involved in cellular interactions between trophoblastic and endometrial epithelial cells, we have established two human cell lines, trophoblastic HT-H and endometrial epithelial SNGM. These two cell types exhibit cell adhesion at their respective apical cell membranes. Molecules involved in this unique cell adhesion were identified by expression complementary DNA cloning and were named trophinin, tastin, and bystin. Trophinin is a membrane protein thought to have self-binding activity and thus mediates homophilic cell adhesion. Tastin and bystin are cytoplasmic proteins required for trophinin to exhibit cell adhesion activity. Trophinin is strongly expressed in trophectoderm of monkey blastocysts. In human endometrium, trophinin is expressed for a limited period in the luminal epithelium at the time expected for implantation. In human placenta, trophinin, tastin, and bystin are strongly expressed in trophoblast and endometrium at the uteroplacental interface at an early stage in pregnancy. All these molecules disappear from the human placenta in the second trimester. The unique expression pattern and cell adhesion activity exhibited by trophinin, tastin, and bystin suggest strongly the involvement of these molecules in the initial attachment of blastocyst to uterus. PMID- 10797942 TI - Steroid-regulated molecular markers of implantation. AB - The endometrium acquires the ability to implant the developing embryo within a specific time window, termed the "receptive phase." During this period the endometrium undergoes pronounced structural and functional changes induced by the ovarian steroids estrogen and progesterone, which prepare it to be receptive to invasion by the embryo. These steroid-induced molecules, when identified, may serve as useful markers of uterine receptivity. In the present article, we provide a brief description of the molecules that have emerged as candidate markers of steroid hormone action in rodents and humans during implantation. PMID- 10797943 TI - Progesterone-regulated gene expression in the primate endometrium. AB - Progesterone action is essential for maturation of the endometrium to a receptive state for implantation in humans and nonhuman primates. The orchestration of progesterone-regulated gene expression is also temporally controlled during the secretory phase based on the limited window for implantation. The genes and gene networks affected by progesterone are likely to involve both activation and repression. Our laboratory has used the rhesus monkey as a model to study the regulation of genes known or suspected to be involved in endometrial maturation. In addition, we have used subtractive hybridization and differential display techniques to identify novel or unsuspected genes that are regulated by progesterone during endometrial maturation. Our studies have led us to propose a working model of progesterone action during the primate secretory phase that includes waves of gene activation and repression that culminate in a receptive endometrium. PMID- 10797944 TI - Implantation in the baboon: endometrial responses. AB - Blastocyst implantation in the baboon usually occurs between 8 and 10 days post ovulation. Changes that occur within this window of receptivity and immediately following implantation can be divided into three distinct phases. The first phase, regulated by estrogen and progesterone, is characterized primarily by changes in both the luminal and glandular epithelial cells in preparation for blastocyst apposition and attachment. The second phase is the further modulation of these steroid induced changes in both epithelial and stromal cells by embryonic signals. The final phase is associated with trophoblast invasion and the remodeling of the endometrial stromal compartment. During the initial phase, the actions of estrogen and progesterone are dependent on the presence of specific receptors. Estrogen up-regulates both its own receptor (ER) and the progesterone receptor (PR), while progesterone down-regulates this expression pattern. However, the pattern of progesterone-induced down-regulation of ER and PR is confined to the epithelial cells and demonstrates a gradient effect from the functionalis to the basalis. What is most intriguing is that the loss of epithelial PR is closely correlated with the establishment of uterine receptivity. Coincident with the changes in ER and PR expression, epithelial cells undergo alterations in their cytoskeletal architecture and secretory profile. These changes can be counteracted by PR antagonist treatment during the luteal phase. Although estrogen and progesterone play a critical role in establishing the initial phase of uterine receptivity, it is becoming increasingly evident that the embryo induces functional receptivity in ruminants and rodents. In our studies in the primate, we demonstrate that chorionic gonadotrophin when infused in a manner that mimics blastocyst transit, has physiological effects on the three major cell types in the uterine endometrium. The luminal epithelium undergoes endoreplication and distinct epithelial plaques are evident. The glandular epithelium responds by inducing transcriptional and post-translational modifications in the major secretory product, glycodelin. The stromal fibroblasts initiate their differentiation process into a decidual phenotype and are characterized by the expression of actin filaments. In phase three, blastocyst attachment to the surface epithelium and subsequent implantation is associated with local remodeling of the maternal stroma, smooth muscle, and endothelium of the blood vessels by the trophoblast. In addition, there is a gradual diminution of the epithelial plaques on the luminal surface although the glandular epithelium remains highly secretory. The most dramatic effect is on the stromal fibroblasts, which in response to embryonic stimuli, differentiate into decidual cells, the major cell type of the gestational endometrium. This differentiation is characterized by the expression of insulin like growth factor binding protein-1 (IGFBP-1) in these cells. The cytokine IL-1 beta is one possible embryonic signal. COX-2 is the rate-limiting enzyme for prostaglandin biosynthesis and transcription of this enzyme in response to the embryonic stimulus (IL-1 beta) results in an increase in prostaglandin biosynthesis in stromal fibroblasts at the site of implantation. Prostaglandins and PGE2 in particular, binds to its specific receptor (EP2 or EP4) and activates adenyl cyclase. The resulting increase in intracellular levels of cAMP can now activate IGFBP-1 gene transcription at the site of implantation. In summary, our studies have demonstrated that chorionic gonadotrophin, when infused into non pregnant baboons during the window of uterine receptivity can induce epithelial responses that are similar to those observed in a fertile cycle. Stromal differentiation is initiated; however, decidualization requires a signal from the conceptus. PMID- 10797945 TI - Embryonic regulation in implantation. AB - Maternal steroid hormones play a critical role in establishing the receptive phase of implantation. In addition to that, the embryo is able to modulate endometrial molecules during the apposition phase (chemokines) and the adhesion phase (adhesion and anti-adhesion molecules). Moreover, the human embryo also exerts a coordinated regulation of endometrial epithelial apoptosis during these implantation phases. In this work, we analyze the embryonic regulation of implantation in humans using an in vitro model. PMID- 10797946 TI - Embryo implantation and tumor metastasis: common pathways of invasion and angiogenesis. AB - Implantation of the embryo is one of the last great mysteries of reproductive biology. There are striking similarities present between the behavior of invasive placental cells and that of invasive cancer cells. In this review, we propose that cellular mechanisms used by the cells of the placenta during implantation are reused by cancer cells to invade and spread within the body. Integrins and other cell adhesion molecules, extracellular matrix and matrix metalloproteinases all appear to be involved and are regulated by the complex endocrine, autocrine and paracrine milieu within the uterus. Angiogenesis is a common feature of both implantation and cancer spread. Endothelial cells also use similar cellular mechanisms during angiogenesis to digest the surrounding matrix, migrate and form new blood vessels. A better understanding of the mechanism of trophoblast invasion will likely lead to insights of various diseases of pregnancy such as preeclampsia. An appreciation of the maternal mechanisms to control this invasive behavior may likewise lead to a better understanding of metastatic cancer cells and lead to better methods to control their growth and spread within host tissues. PMID- 10797947 TI - [Vaginal injuries during coitus]. AB - Based on a prospective study on a continuous series of women admitted on emergency for post-coitus hemorrhage. The authors present an epidemiology and prognosis of vaginal injuries in to coitus in the Dakar University Teaching Hospital. Over a period of 33 months (October 1991 to December 1994), 106 cases were recorded, giving an incidence rate of 32 cases per year. The average profile is that of young women (26 years), usually during the post-partum period (41.5%) or in the second phase of menstrual cycle (32%). The main influencing factors found included: chronic vaginal infection (74%), position during coitus (especially that dorsal decubitus with hyperflexion and abduction of the lower members (56%), sexual abstinence of 3 months on average (21 days to 2 years), and use of aggressive product "aphrodisiac" for vaginal mucus. The damage was most often located at the bottom of dead end of Douglas and could be estimated at 5 cm long and 5 mm deep. The immediate prognosis was favourable, after stitching with absorbable thread, healing was obtained within 8 to 15 days. Given a retrospective view about 2 to 5 years, morbidity was represented by 3 recurrent cases and 13% after effects of the deep dyspareunia type. PMID- 10797948 TI - [Ethmoid tumors. Experience of the otorhinolaryngologic clinic of the University Hospital Center of Dakar, 20 cases]. AB - 24 cases of ethmoid tumors collected between January 1st, 1990 and June 30th, 1996 were reported. The analysis of epidemiological, clinical and therapeutic aspects showed the following elements: the average age was 25 years with an equal repartition between men and women, a delay in consultation with an average of 2 years, 41.7% of patients consulted mostly for non rhinological signs, on the therapeutic aspect, surgery was performed on 95.8% of the patients with ethmoidectomy by external way. In 50% of cases, the removal was extended to one or many sinuses, 61.2% of tumors were benign to the histological analysis, one patient died immediately after operations. In the short run, we observed 2 cases of persistence of the carcinoma. In the long run, a recurrence of mucocele was noticed. Because of late consultation, limits of preoperative assessment notably for radiology, therapeutic approach is difficult, the improvement of technical infrastructures, the use of scanner in particular, should allow a better codification of the therapeutic indications. PMID- 10797949 TI - [Burkholderia cepacia isolation and characterization from hospital infections]. AB - The evolution of reanimation and functional exploration techniques has led to and explosion of nosocomial infections. They are prevailing in Intensive Care and Neonatal Units. This study deals with B. cepacia strains isolated in 1996 in a pediatric hospital of the Dakar University Hospital Center, following the installation of tracheo-bronchial exhausters which are used for obstruction removal among children. The 44 B. cepacia strains examined come from 42 blood cultures done among 29 boys and 13 girls aged between 5 days and 7 years, and from 2 exhausters. After identification by API20 NE (bio Merieux), a standard antibiogram, a 3 characters biotyping (O.N.P.G., esculin, nitrate reductase) and a study of the polymorphism of the DNA enzymatic restriction profile obtained by an pulsed field electrophoresis are performed on the isolates. The contamination come from the exhausters. All the strains produce an orange-colored yellow pigment. Only an O.N.P.G. (+), nitrate reductase (+) biotype was identified. The antibiotic susceptibility profile is almost pathognomonic for the 44 tested strains: sensitivity (100%) to ceftriaxone, to ceftazidime, to aztreonam: to contrimoxazole (96%) and to chloramphenicol (91%). Search for widen spectrum beta lactamses and antibiotics resistance plasmids was negative. However, those strains that are multiples resistant, discharge others 8.1. isofocal point beta lactamases. The R.F.L.P. study demonstrated a unique profile. The B. cepacia transmission is the result of the installation of medical reanimation equipment that are not well taken care of. The nosocomial infections ascertained so far are ordinary bacteremias. Strain's phenotypical and genotypical identification shows the presence of only one clone. To overcome there nosocomial infections, hygienic measures have to be reinforced. PMID- 10797951 TI - [Clinical criteria for oral pathology in a history of HIV infection]. AB - Oral lesions have been shown to be the first clinical symptom in HIV infection by a large number of studies. A cross prospective study in Infectious Disease and Dental Service of Fann Hospital, the objectives were to evaluate the prevalence of oral lesions and to determinate correlation between oral lesions and CD4 cells count in HIV+ patients. 1.645 patients were examined, 352 were eligible. The prevalence of oral lesions was 62.7% in HIV+ patients. Among those 135 HIV+, 56.15% had a rate of CD4 < 200/mm3. In addition with studies presented in Third International Conference of Oral Lesions in HIV Disease (1996 May), oral lesions can be used as indicators for progression of HIV Disease. PMID- 10797950 TI - [Community nutrition strategy project: an innovation in community health]. AB - The strategy of the community nutrition project is based on the utilization of the community development structures to deliver the nutrition services. These structures, represented in Senegal by youth associations, women groups, GIEs and NGOs, are part of the decentralization process, and as such play an important role in health and health development activities in poor urban districts. The Community Nutrition Project (CNP), funded for five years by the World Bank, German Cooperation (KFW), World Food Program (WFP) and the Senegalese government aims to halt further deterioration in the nutrition status of the most vulnerable groups in the poorest urban districts of Senegal. All nutrition services and particularly the IEC services have been entirely contracted out the first year to 76 GIEs involving 323 unemployed persons, operating as micro-enterprises "MIC" and 17 "GIEs" of unemployed physicians, pharmacists, and social workers for a total of 34 persons, organized as "maitre d'Oeuvre communautaires "MOC", in charge of the supervision tasks. Each community nutrition center recruits and monitors every six months 460 to 600 beneficiaries composed of women at six months of pregnancy, lactating mother of children under 6 months, and a group of children aged from 6 to 35 months old. An average of 87% of registered children in the nutrition centers are weekly or monthly weighted. Thus the proportion of malnourished children in cohort of children followed from January to July 1996 has decreased from 70% to 25% within six months. The malnutrition rate has been reduced up to 65% after six months. PMID- 10797952 TI - [Well water pollution in the Khombole district: research on the contamination by organochlorine pesticide residues and organic substances (feces)]. AB - The study realized in the district of Khombole (SENEGAL) has permit to estimate the contamination levels of wells waters used by the populations. The research and the dosage of the organichlorine pesticide residues, nitrites and nitrates and microbiologic analysis have been done on 19 wells chosen after a drawning of lots. The organochlorine pesticide residues which have been found prove that the wells are permanently exposed to these chemical substances which don't constitute nevertheless a major risk for the populations health. The results of our research proved also that there is a real risk of intoxication with the nitrogen oxides. In effects more than 50% of the wells have revealed nitrates contents up to the indicative value (25 mg/l). As for the nitrates, with a few exceptions (5/18), the contents are superior to the authorized norm (0.1 mg/l). By another way the bacteriologic analysis has revealed in the one hand a DBT (Total Bacterian Count) up to 10,000 germs/l for all the wells, and in the other hand the presence of Escherichia coli, Enterococcus faecalis that confirm a faecal contamination. PMID- 10797953 TI - [Effects of spatial orientation on attention reaction time during pointing movement towards visual direction]. AB - In the present study, it was consequently proposed to investigate how the spatial orientation of attention made by the explicit and implicit components of advance information, affected the reaction time (RT) performances. Subjects performed a simple RT task with an orientation cue and two choice RT situations, the one with a neutral cue and the other with a primed cue. The motor task to be performed consisted of pointing towards a visual target. The mechanisms involved in the orientation of attention were studied on the one hand by analysing the effects of the preparatory signal, which explicitly oriented the subjects' attention towards the forthcoming target position, and on the other hand, by examining whether the reinforced probability of warning stimulus(WS) and imperative stimulus (IS) being compatible affected the subjects' performances, i.e., whether they detected and made use of the information implicitly conveyed that there existed a bias in favour of the probability that IS would appear in the direction predicted by WS. The results of this study show that the subjects shortened their RTs by using both the explicit(auditory) and the implicit(probability-related) information with which they were provided prior to performing the movement. These data on the effects of the oriented auditory signal on the reaction times indicate that in our experiments, by orienting their attention depending on the position of the stimuli, the subjects were able to reduce their reaction times. The significant correlation found to exist here between the level of probability associated with the position of the WS and the choice RT values indicates that also the implicit information as to the WS-IS compatibility was taken into account by the subjects in all the situations. PMID- 10797954 TI - [Nitric oxide and isolated aortic contraction in a pregnant hypertension model by the inhibition of nitric oxide synthase in pregnant Wistar rats]. AB - During gestation endothelium induces decreases in vascular responses to vasopressor agents but endothelium disease is followed by hypertension and enhanced vascular reactivity during preeclampsia. In a rat model of preeclampsia induced by NO synthase inhibition we study here isolated aortic contractions. From day 13 of gestation 2 groups of Wistar female rats were fed control (C) or nitro-arginine enriched diets (0.063%, i.e. 30 mg/kg/d) (treated) (T). On gestational day 20 systolic blood pressure (SBP, mmHg) is measured by tail cuff method and isolated thoracic ring aorta contractions are studied after depolarisation (KCl 60 mM) or norepinephrine (cumulative concentrations 10-9 M-10 5 M). After chronic NOS inhibition, hypertension develops: SBP is 154 +/- 2.17 in T and 116 +/- 3.75 in C, p < 0.01 and significant proteinuria (mg/d) appears: T, 63.4 +/- 21.6 versus C 3.08 +/- 0.48, p < 0.01. NO synthase inhibition in treated rats impairs the depressed contractile response obtained in the presence of endothelium in control rats but addition of L-arginine suppresses the effect of nitroarginine. Taking in account our results and those described in literature it appears that L-arginine treatment could ameliorate some pathologic pregnancies. PMID- 10797955 TI - [Characterization of entomopathogenic Bacillus samples isolated in Senegal and study of their toxicity for malaria vectors]. AB - A screening program developed in Senegal to isolate new strains of entomopathogenic Bacillus has led to the isolation of 194 strains of Bacillus thuringiensis and 9 strains of Bacillus sphaericus from various sites and insect samples. The characterization of these strains regarding their H serotype, their crystal composition and their toxicity against mosquitoes (Culex pipiens, Aedes aegypti and Anopheles stephensi) has led to the isolation of 27 mosquitocidal strains. As malaria is an important public health problem in Senegal, these strains were more completely characterized looking for their toxicity against the two major malaria vectors in Senegal: Anopheles gambiae and Anopheles arabiensis. PMID- 10797956 TI - [Prognostic value of cytokines and HLA markers in patients with cerebral malaria in Senegal]. AB - 41 patients senegalese patients suffering from clinically defined severe malaria were studied in the intensive medical care unit of the Hopital Principal in Dakar, Senegal. All of these individuals lived in Dakar, an area of low and seasonal Plasmodium falciparum transmission. In this study, we aim to determine in one hand, the cytokine levels of TNF-alpha, TNF-alpha sRI, TNF-alpha sRII, IL 2 sR, IL-6, IL-6 sR, and IL-10 to evaluate their prognostic value in the course of the disease; in the other hand, the influence of the HLA-DR alleles in the susceptibility to get severe malaria. At the day of admission (day 0) and 3 days later, one or two blood samples were collected for each patient to assess different biological parameters. Plasma samples were tested for cytokines cited above by ELISA (Medegenix EASIA kits) and DNA samples for HLA-DR by PCR-SSP genotyping. The concentrations of all the cytokines and/or their receptors were significantly increased at day 0 in the patients who died (TNF-alpha = 455 +/- 480 pg/ml, IL6 = 511 +/- 396 pg/ml) and decreased rapidly in the patients who survived from the disease (TNF-alpha = 354 +/- 629 pg/ml, IL6 = 453 +/- 706 pg/ml). A fatal issue seems likely related to the age of patients (20 +/- 12 years for surviving patients and 31 +/- 16 years for patients who died) and the kinetic of the cytokines. Significant differences were observed (pc < 0.001) between patients with severe malaria and a control group for the following HLA alleles: DR3, DR10 and DR13. The HLA-DR13 allele was found positively and highly associated with severe malaria. PMID- 10797957 TI - [Spinal neurinomas and neurofibromas in Senegal]. AB - Authors present 13 cases of spinal neurinomas and neurofibromas out of 418 non- pottic, non-traumatic medullary compression operated between 1965 and 1995. All cases were histologically documented. Neurinomas and neurofibromas represent 3.1% of all non pottic, non traumatic medullary compression and 12.8% of tumour medullary compression operated in our service. The illness started with radicular pains (6 cases) and spinal ache (5 cases) and most of the time, the medullary compression diagnosis is completed at the spasmodic paraplegia status. No patient has done any C.T. scan but in the other hand a myelography has been completed in all cases, resulting in the blockage of the contrast product which in 4 cases was of an epidural type, and in 2 cases of intra dural type. All patients were operated through a posterior approach, which ended in 10 cases to a total removal, and in 3 cases to a partial one. 7 extradural localizations were noticed and 3 neurinomas, 3 neurofibromas and 1 neurofibrosarcoma were histologically identified. A post operative follow-up was done on 11 patients and a satisfactory result was obtained on 6 of them. In 2 cases, the neurological status remained unchanged. In 2 cases a deterioration occurred and a patient with neurofibrosarcoma passed away. PMID- 10797958 TI - [Evaluation of parasitic risks related to the revitalization of the Ferlo fossil valley (Senegal)]. AB - In order to assess the parasitic risks related to Ferlo's valley water launching, a study has been carried out from september 15 to october 10, 1996 in 12 villages. Four villages surrounding the Guiers lake, four lower within lower Ferlo already water launched 7 years ago, and four villages within upper Ferlo not water launched. Malaria strikes at hypoendemic level in the villages surrounding the Guiers lake (p.i = 6.2%) and at mesoendemic level in the area stretching along lower (P.I = 37.6%) and upper Ferlo (P.I = 34.3%) The prevalence rate of urinary bilharziosis is 0.002% in the first area, 1.3% in the second one and 14.5% in the third area. In these areas, intestinal parasitosis were prevailing respectively at rates of 38.2%, 36.4% and 22.2%. Although, there is no reason to fear immediately a worsening of the epidemiological situation due to Ferlo's valley revitalisation project, nevertheless, appropriate steps should be taken right now aiming at tacking the extension of conditions related to hydric medium. PMID- 10797959 TI - [Risk factors for low birth weight: influence of maternal age, parity, gestational age, nutritional status and maternal pathology]. AB - A retrospective study carried on between april 1st and september 31st 1997 has helped in collecting. 69 cases of low birth weight new-born (weight < 2500 g) at Abass Ndao hospital center in Dakar. The above population has been compared to 79 eutrophic new-born of mean birth weight equal to 3047.7 +/- 311 g (witnesses). The goal of this study is to appreciate the relationship between the maternal age, the number of the gestation, the parity, the nutritional status, the maternal pathologies during pregnancy and the low weight at birth. There was no difference between both groups as regards to the mean age (p = 0.44), the mean number of gestation (p = 0.7) and the mean parity (p = 0.48). On the other hand, the weight of the mother is smaller as for the group of low birth weight but the mean body mass index stand at normal in both groups. The pathologies during pregnancy period were obviously more frequent in the group of low birth weight new-born. There is a real need to insist on the preventive measures to be taken and the treatment of the maternal pathologies during pregnancy. PMID- 10797960 TI - [Epidemiologic and obstetric aspects of adolescent pregnancies in the hospital milieu]. AB - Adolescence is a period of change where the person is seeking an identity. This period is characterized by an early sexuality with early pregnancies and risky deliveries. The objectives of our study are to show the epidemiologic aspects of adolescent pregnancy and to identify its obstetrical factors. It is a prospective study held in one year, from August 1st 1991 to July 31 1992 including all pregnant adolescents and those coming for delivery at the Obstetrics and Gynecology clinic at the University teaching hospital Le Dantec. The prevalence of adolescent pregnancy is 3.7% and 84% are primiparous. The non-educated represent 65% and 55% are married. Early sexuality and early pregnancy are mostly seen among Pulars. The complications during pregnancy are toxemia and its accidents (17.6%), premature rupture of membrane (4.4%) and anemia (2.5%). Adolescent who had not received prenatal care represent 11% and 38% had not received enough care. Normal delivery happened in 67.5% and the C-section represented 7.5%. 3% of the adolescents had done unsafe abortion. No maternal death was observed. 89.1% live births were reported, however, 18.4% needed intensive care. Because of the obstetric and neonatal problems, early pregnancy should be prevented through a active collaboration of divers partners. PMID- 10797961 TI - [Clinical aspects of adult schizophrenia in the psychiatric service at the Yalgado Ouedraogo National Hospital Center of Ouagadougou (Burkina Faso)]. AB - Schizophrenia is a chronic mental disease severe and disabling, which affects young adults. It is a frequent disease which, due to its disabling nature concerns as well the society, the family as the individual. We have gone throughout the files of 61 inpatients in the psychiatric department of the National Hospital Centre Yalgado Ouedraogo of Ouagadougou, using the diagnosis of schizophrenia according the criteria of the World Health Organization. The aim of our study was to draw up the socio-demographic profiles and the symptoms of these schizophrenic patients. Our results emphasize the importance of delirious productions (80.3%) and disorders of the perceptive field, particularly auditive hallucinations (51.5%) in the semiology of the disease. On the diagnostic plan, paranoid forms prevailed (74.5%) in our series. The long course of the schizophrenia process preceding the consultation often makes the diagnosis easy at the patient's arrival. Clinical forms with noisy symptomatology, usually not much tolerated by families, often ask for medical care, sometimes after unsuccessful traditional therapy. PMID- 10797962 TI - [Staphylococcus aureus purulent pleurisy in children. Experience of the Albert Royer Hospital for Children of the Fann University Hospital Center in Dakar]. AB - The authors report here by a retrospective study 58 cases of Staphylococcus aureus empyema at Albert ROYER child hospital located in the Fann University Teaching Hospital of Fann between January 1st 1992 and December 31, 1995. In this study staphylococcus aureus is the bacterium involved in pleural effusions of the children (54%) a long way ahead Streptococcus pneumoniae (16%). Infant less than 30 month is more affected (86%). The average age of the patients is 16.8 month +/ 16.6. The resistance of the germ to usual antibiotics, the precariousness of the research field and mechanical complications linked to the outpouring explain their seriousness. The treatment lies upon an adapted antibiotic and bactericidal therapy associated to closed chest tube drainage. PMID- 10797963 TI - [Scabies complicated by acute glomerulonephritis in children: 114 cases observed in two years in a pediatric service in Dakar]. AB - We performed a retrospective study during 2 years in order to determine the clinical and epidemilogic characteristics of 144 cases of scabiosis with acute glomerulo-nephritis in Dakar. The 144 cases of scabiosis complicated with glomerulo-nephritis represented 56.6% of the whole 205 cases of scabiosis observed in the same period. The most important part of the cases was observed between November and January. Usually children came from poor and numerous family. The scabios lesions were mostly generalised and the infection was recent. The main features of the kidney disease were: oedemas(93.2%), high blood pressure (89.1%) and haematuria(54.70%). Cardiac and neurological involvement were the main complications. The global mortality was very low(1 case). The evolution was good in 97% of the case with complete cure. PMID- 10797964 TI - [Autosomal dominant polycystosis in the hospital milieu in Dakar (Senegal)]. AB - Autosomial dominant polycystic kidney disease (ADPKD) is an hereditary affection transmitted in a autosal dominant pattern with variable penetrancy. Diagnosis is based upon ultrasound examination and/or familial history. The authors present a 8 years retrospective hospital study in order to precise the epidemiological and clinico-biological aspects of ADPKD in Dakar. So 23 patients were studied. The mean age at the diagnosis was 46 years. They were 9 men and 14 women (sex ratio 0.64). A past history of familial nephropathy was found in 52.2% of cases. The circumstances of diagnosis were dominated by lombar pain (52.2%) and hypertension (17.4%) but at examination, blood pressure was high in 34.8% and represent the main factor of worsening the disease. Enlargement of kidney was found in 47.8% and hepatomegaly in 13% of cases. Anemia was particularly common 56.5%. The most frequent extra renal manifestation was the liver cysts and occurs mainly in women. National prevalence of ADPKD and the specific gene should be precise in further studies. PMID- 10797965 TI - [Epidemiological and clinical aspects of pancreatic cancer in Senegal]. AB - The objective was to verify if the introduction of ultrasonography in our practice has entailed modifications of the epidemiology and the clinic of pancreatic cancer in Senegal. We have performed a retrospective study of cancer of the pancreas diagnosed on the basis of clinical signs, ultrasonography, surgical findings and necropsy from March 92 to October 97. The chi-square test according to Mantel-Haenszel has been used to compare qualitative variables. The routine practice of ultrasonography has allowed to establish the diagnosis of 107 cases of pancreatic cancer within 5 years and half. The sex ratio was 1.05 and the average age was 60 years +/- 13 without significant difference according to the sex. The mean period of diagnosis was 5 months. However the classic form associating thinness and cholestatic jaundice or palpable abdominal mass dominated the clinical symptoms (74.5% of cases). A very deep alteration of the general status has been noticed in 25.5% of cases. At surgery, according to the TNM classification, 22% of our patients were at stage II and 78% from stage III to IV. In Senegal, the prevalence of pancreatic cancer is increasing since the introduction of ultrasonography. This technique has shown that the male predominance is less than reported in previous studies. Nevertheless the diagnosis remains late. As far as we cannot identify risk groups, recent appearance of digestive disturbance even atypical after 50 years old, is enough to suspect the diagnosis. PMID- 10797966 TI - [Malignant kidney tumors in adults in Senegal: diagnostic and therapeutic problems]. AB - The aim of this study was to analyze problems posed by adult renal malignant tumors both in the diagnostical and therapeutical viewpoints. We have performed a retrospective study including 48 cases of confirmed renal cancer. Have been included adult patients who presented a cancer of the kidney operated or not. Studied parameters were age, sex, clinical signs, complementary explorations i.e. biology, renal ultrasonography, IVU, Computerized tomography, the operative indication and the prognosis. We have used Robson's staging to classify our patients. The average age was 51 years with extremes of 18 and 83 years. However 60% of them were less than 50 years. Adenocarcinoma was the main histological form encountered. It has been found in 93% of cases. The flank mass has been the mode of discovery in 70.8% of cases. The classic symptomatic triad associating hematuria, pain and the renal mass has been found in 20.8% of cases. Ultrasonography and Intravenous urography (IVU) have allowed to pose the diagnosis in 93.75% of cases. Patients were often referred very late with an average delay of consultation of 14 month. So, 25% of them presented with metastasis at the moment of the diagnosis. Radical nephrectomy was performed in 60% of cases. 40% of patients have not been operated due to the advanced evolution of the tumor. The global mortality to 1 year was estimated at 38%. We conclude that renal malignant tumors in the adulthood, in our practice, presents some particularities. Indeed, patients are referred late when clinical symptoms are sufficiently evocative to suspect the diagnosis. At these advanced stages, there is no curative therapeutic. So, a precocious diagnosis is necessary in order to propose to patients radical nephrectomy which remains the only efficient therapeutical procedure of the localized renal cancer in the adulthood. PMID- 10797967 TI - [Long term evaluation of results of endodontic treatments of dental pulp necrosis (74 cases obturated by the monoconal technique]. AB - Various factors may affect the outcome of root canal therapy. In this study 74 endodontocally treated pulpless teeth were evaluated: (43 with periapical radiolucency and 31 without periapical lesions). These 74 non surgical treatment were performed in 60 patients (26 men and 34 women) by undergraduated students. The overall success rate was 71.62%; the rate of success for cases with no periapical radiolucency exceeded 80% whereas only 62.8% of the cases with perapical radiolucency had undergone partial resolution or complete resolution. Only 17.1% of lesions demonstrated complete radiographic resolution 12 to 16 months after the endodontic treatment. Reasons that may explain the low success rate are lake of asepty, root canal preparation and root canal filling technique. PMID- 10797968 TI - [Indications and prognosis of forceps delivery at the University Hospital Center of Dakar]. AB - Based on a study of 382 cases registered in a five years periods (1992-1996), the prevalence of forceps delivery is evaluated at 16 per 1000 deliveries. Averagely these are young (24.5 years), primipare (63%) at term and having problems of abnormal labour. The mains indications are: assistance of expulsion (47.1%), severe foetal distress (15.6%), scarred uterus (15.2%) and eclampsia (8.9%). The most frequent applications are the occipito-pubic grasp (51.3%) and left anterior occipito-iliac grasp (34.8). The maternal morbidity prognosis (6%) is represented by cervical or vaginal traumas and infections. The neonatal prognosis is marked by neonatal mortality rate of 50 per 1000 of life deliveries and by a morbidity rate of 3.4% essentially represented by cutaneous lesions. PMID- 10797969 TI - [Duodenal diaphragm in children (two cases)]. AB - Duodenal diaphragm is a rare cause of intestinal obstruction. Two pediatric cases are reported in which vomiting of bile-stained material and distension of the epigastrium were the main clinical signs. Roentgenograms and sonograms were highly suggestive. Duodenal bypass was easy to perform and effective. Nevertheless, it must be kept in mind that associated anomalies are frequent and may be severe. PMID- 10797970 TI - [Adult T-cell leukemia-lymphoma due to HTLV 1. Two cases of the acute form at the Principal Hospital of Dakar]. AB - HTLV1 virus is a retrovirus that has been endemic in Africa. It is the responsible for tropical spastic paraparesis and adult's T Cell leukemia lymphoma. Few cases of adult T-Cell leukemia-lymphoma have been described in Africa, contrary to Japan and Caribbean. Were are reporting two cases of acute adult T-Cell leukemia-lymphoma which characterised by blood lymphoma signs, tumoral nodes and extranods lesions, hypercalcemia and positive retroviral serology of HTLV1 virus. The prognosis of these acute forms was bad after a six month survey approximately. The treatment is disappointing. Investigations of this affection must be carried out in every patient who presents lymphoma manifestations all the more because they are associated with hypercalcemia. PMID- 10797971 TI - [Idiopathic serous retinal pigment epithelial detachment (a case report)]. AB - Retinal pigmented epithelium detachment is frequently associated to age related macular degeneration or others maculopathies. More rarely no cause can be found, particularly in young patients where it constitutes an entity named "Idiopathic serous detachment of the retinal pigmentary epithelium". We report such a case. We insist on the clinical and angiographical characteristics which allows a differentiation from the idiopathic central serous chorioretinopathy with which it can be confused. Last, it is important to point that for the time being no treatment can shorten the evolution. PMID- 10797972 TI - [Peyronie's disease and peripheral neuropathies]. AB - The association between Peyronie's disease and diabetes mellitus or Dupuytren's disease is well described in the literature with a certain frequency. Through our 10 patients with Peyronie's disease, when have noticed that only one of them is undergoing diabetes mellitus and no one has Dupuytren's disease. But 7 of them presented clinical and/or electromyographical signs of peripheral neuropathy. So after clinical examination in neurology and urology, we performed measurement of motor conduction velocity of tibial posterior nerves and peroneal nerves, then, we measured sensitive vetocity of sural nerves and dorsal nerve of the penis and we studied the cortical somesthesic evoked potential of the internal hontous nerve and the sympathetic evoked response of the skin. So we hypothesize that peripheral neuropathy with or without autonomic disorders might be an etiological co-factor of Peyronie's disease. PMID- 10797973 TI - [Spinal cord injuries in Senegal: 16 cases]. AB - Penetrating spinal cord injuries (P.S.C.I.) are rarely described in Sub Saharian countries in spite of an increasing number of wars. To study epidemiology management and prognosis of P.S.C.I. in Senegal, population of 16 patients collected from Fann Hospital in Dakar has been studied. 9 cases were related on gunshot or shrapnel injuries and 6 were stab-wounded. 8 came from war practice and 7 from civilian practice. The point of entry was at the posterior or lateral part of the body and continuous leaking of cerebral spinal fluid from this point was founded only in one patient. Patients showed a clinical picture of a complete spinal cord section syndrome, 3 spinal cord hemisection Brown Sequard syndromes, 3 cauda equina syndromes and 1 monoradicular syndrome. Spinal X-rays or myelography may lead to an accurate evaluation of the extent of bone tissue destruction. Anatomical evaluation of roots and spinal cord lesions were more difficult when C.T. scan or R.M.I. is not available. Penetrating spinal cord injury with foreign body included or myelography stop or showing cauda equina syndrome should be operated on. 9 of our patients has benefited of spine surgical posterior approach (laminectomy). Immediate vital prognosis is good regarding the fact that visceral associated lesions were rare (2 cases). Functional recovery is fair only 46.6% of patients expressed partial or complete recovery. Prognosis factors such as injuring agent and initial neurological status has been discussed. Prognosis of penetrating spinal cord injuries could be improved by immediate and multidisciplinary management. PMID- 10797974 TI - [Cardiac thyrotoxicosis and left ventricular thrombosis, a case report]. AB - Authors report about a left ventricular thrombus as a complication of thyrotoxic heart disease, on a 45 years old woman. Clinical state featured signs of thyrotoxicosis, global cardiac failure and hypertension. The electrocardiogram showed a left axis deviation, the chest X-ray a cardiac enlargement (cardio thoracic index = 0.55) on behalf of left movement of heart walls and severe left ventricule dysfunction (Ejection fraction about 18%). This exam also found a big thrombus at he left ventricule apex. The treatment disappearance of the thrombosis and improvement of cardiac failure signs and echographic parameters. Authors discuss circumstances leading to thrombosis in thyrotoxic heart disease, and the usefulness of anticoagulant drug therapy. PMID- 10797975 TI - [A left paraduodenal hernia--a case report]. AB - The left paraduodenal hernia obtains when the small intestine goes through the left paraduodenal fosset. It (the hernia) develops on the left, at the back of the descending mesocolon. The present authors study one case looking at its pathogeny and the clinical and therapeutic problems posed by such a variety of congenital internal hernia. The authors emphasize, in particular, its morbid associations with the volvulus of the pelvic colon. Finally they highlight the importance of a through and careful exploration of the abdominal cavity during any laparatomy following an occlusive syndrome. PMID- 10797976 TI - [Drug resistant epilepsy in Senegal: therapeutic, clinical and socioeconomic factors]. AB - The aim of our study was to emphasize factors which support pharmaco-resistance in Senegal. For this purpose, 23 patients with partial or generalized epilepsy were studied, after determining the plasmatic concentration of the antiepileptic drugs. The aetiologies were numerous: encephalitis, injuries, neonatal encephalitis. All patients were under traditional treatment before coming to the hospital. So antiepileptic drugs were taken a long time after the beginning of epilepsy later, they were Phenobarbital, Carbamazepine, Phenytoin, and Valproic acid. Only seven patients had sufficient plasmatic level of the antiepileptic drug. The low socio-economic conditions of patients which limit the choice of the most adapted drug in each case, is one of the most important reason of the pharmaco-resistance. Another factor of the pharmaco-resistance is cultural and is linked with the absence of notion of chronic disease necessitating long and regular treatment in senegalese traditional society. PMID- 10797977 TI - [Epidemiological, clinical and therapeutic aspects of severe malaria in adults in the infectious disease department of Central University Hospital of Dakar]. AB - To assess epidemiological, clinical and therapeutic features of severe malaria among adults in Dakar (Senegal), we carried out a 5-year retrospective study in Infectious Diseases Ward (January 1992-December 1996). Over this period, 222 cases of severe malaria were included according to WHO definition criteria, 120 of them (54%) being adult patients. Monthly distribution of cases showed 2 peaks, on October and November. Most of the patients were males (sex-ratio = 2.1) and lived in urban area (91.7%). The mean age was 28.9 years (range = 16-73 years). Clinically, all of the cases presented with stage II coma. Association existed with convulsion (20%), severe anaemia (29.2%), renal failure (19.2%), hypoglycaemia (17.5%) and jaundice (34%). Patients were treated using quinine intravenously. Case fatality rate reached 26.7%, indicating life-threatening potential of malaria in adults living in urban area. PMID- 10797978 TI - [Anti-inflammatory activity of the bark of Khaya senegalensis (A Juss). Preliminary research of structure/activity relationship]. AB - Some anti-inflammatory tests based on the carrageenan induced oedema rat paw method have been carried out with a freeze-dried aqueous extract and an etheropetrolic extract of Cailcedrat (Khaya senegalensis) barks. Then, a preliminary chemical study to find a relation structure/activity has been done from the etheropetrolic extract, by using various chromatography technicals. The achieved results have shown a decrease of the oedema by 14% when the aqueous extract is used at 1 g.kg-1, 21.5% and 62% with the etheropetrolic extract used at the respecive doses of 0.5 g.kg-1 and 1 g.kg-1. As for the chemical study, it allowed the separation of two compounds which probably take part in noticed activity and are supposed to be methyl angolensate and methyl hydroxyangolensate. PMID- 10797979 TI - [Analysis of implant position after envelope technique in "white" cataract surgery]. AB - Capsulopunture in cataract surgery doesn't allow insertion of intraocular lens in the capsular bag. Envelope technique is a easy way for this insertion. The goal of our study was to analyse the position of the intraocular lens in the capsular bag, after performing envelope technique in eyes suffering from "white" cataract. We realised on 25 eyes with "white" cataract, envelope technique. GALAND method (one superior linear capsulotomy) was realised in 15 cases and double linear capsulotomy (superior and inferior), was realised in 10 cases. The mean age of our patients was 55 years with two extremes (17 and 70 years). The mean visual acuity was 14/20. After verification, in 91.636% eyes, intraocular lenses were in the capsular bag. In 2 cases, we found a decentration. Envelope technique can lessen risks of capsular zip. It protects endothelium cells and is helpful for the insertion of the intraocular lens in the capsular bag. Envelope technique is well adapted to the surgery of "White" Cataract. PMID- 10797980 TI - [Sensitivity to cotrimoxazole of bacteria isolated at the Central University Hospital of Fann, Dakar]. AB - This study concern a survey of bacterial resistance to cotrimoxazole; 510 strains of Enterobacteria (167), Vibrio cholerae(206) and Staphylococcus aureus(137) were tested by disc diffusion and agar dilution methods. An interview was conducted with 86 health personals to appreciate the influence of prescription. Staphylococcus aureus were the most susceptible bacteria (13% of resistance), and Vibrio cholerae the most resistant (95%). Related to the gender, Enterobacteria present 43 to 72% of resistance. The data of interview show a very frequent use of cotrimoxazole, related to the disponibility and the accessibility of this drug. PMID- 10797981 TI - [Comparative study of the antithrombotic effect of aspirin and Bay U3405, antagonist of a thromboxane A2 receptor]. AB - The antithrombotic activity of aspirin (acetylsalicylic acid), inhibitor of the synthesis of thromboxane A2 was compared to that of Bay U3405, a thromboxane A2 receptor antagonist. A model of experimental thrombosis induced by laser in guinea pig mesenteric arteries was used. Aspirin exhibited a dose-dependent antithrombotic activity with and inhibitory effect of 70% at the high dose of 100 mg/kg while Bay U 3405 was efficient at the dose of 10 mg/kg with a percent of inhibition that did not exceed 55%. These results showed that: first, the inhibition of the activity of thromboxane A2 by an antagonist was not more important than that by aspirin and second, the thromboxane A2 way was well implicated in the thrombosis in the guinea pig, suggesting that this species was a good model for the study of the mechanisms using the prostaglandins way in arterial thrombosis. PMID- 10797982 TI - [Pneumocystosis in HIV infected patients presenting with acid-fast bacilli negative pneumopathy at the Central University Hospital at Dakar]. AB - Pneumocystosis is an opportunist parasitic disease which occurs currently at Europe and United States in HIV infected patients. In Africa, the disease is not current. Pneumocystosis has been detected in the Fann medical universitary center at Dakar from HIV infected patients with acido alcoholo resistant bacilli negative pneumopathy. Analysis of broncho alveolar liquid(BAL) of 29 patients after Giemsa and Blue of Toluidin O staining allowed isolating of two cases of pneumocystosis. A man and a woman were the patients. They were HIV1 positive with at X ray bilateral interstitial syndrome. The CD4 lymphocytes count of the one was lower than 200/mm3 and for the other it was higher than 200/mm3. PMID- 10797983 TI - [Risk factors for lymphedema of the arm after mastectomy for breast cancer]. AB - Postmastectomy lymphedema of the arm is frequently associated to different factors including axillary node involvement and local and regional treatment of breast cancer. Our aims was to identify risk factors of postmastectomy lymphedema. From a retrospective analysis of 735 breast cancers treated in our institute, we found 61 lymphedema of the arm. We then describe our study population and identify by univariate et multivariate analysis the factors significantly associated to the disease. The majority of the patients were young black African female found to have locally advanced breast cancers (88% of T3 et T4 UICC 1988), inflammatory diseases (46% of PEV 2 and 3 of Gustave ROUSSY Institute classification of inflammatory breast cancers). Ulceration is found in half of the patients, metastasis in 20%. The patients first underwent chemotherapy mainly with cyclophosphamide alone (56%). Only 59 patients (8%) had preoperative radiation. Surgery consisted mainly in modified radical mastectomy and lymph node dissection (95%). Residual disease is left in 50% of the cases. Only 35% had post-operative chemotherapy and 9% postoperative external beam radiation therapy. From that population, during the follow up, 61 patients were found to have postmastectomy lymphedema. The disease was asymptomatic in 60% of the cases and painful in 26%. 30% of all the patients spontaneously partially regressed. From univariate analysis we found 7 factors associated with lymphedema: The big size of the tumor (p = 0.005), clinically involved axillary lymph nodes (p = 0.001), metastatic disease (p = 0.0046), traditional or inadequate surgery out of the Institute (p = 0.001), lack of post-operative chemotherapy (p = 0.002), postoperative external beam radiations (p = 0.005), relapse (p = 0.002). From logistic regression analysis three independent factors were found: clinically involved axillary lymph nodes (p = 0.0267), metastasis (p = 0.0002) and local or regional relapse (p = 0.0405). In our practice we found that advanced disease, treated by traditional healers or surgery nurses who had relapsed after mastectomy and external beam radiations without chemotherapy have higher risks of lymphedema. PMID- 10797985 TI - [Contribution of medical imaging in osteoarticular manifestations of sickle cell anemia in the child]. AB - Drepanocytosis is one of the qualitative hemoglobinopathies which are the most widespread in the word. In a retrospective study on 16 drepanocytary patients observed at the Bordeaux Children's hospital, 7 patients showed medullar hyperplasia, 10 patients showed vaso-occlusive, and 4 patients showed infections. Medullar hyperplasia: bone demineralisation was the current appearance (47%), cortical thinning down (40.5%). They touch long bone. Thrombosis: bone demineralisation was the current appearance (86.3%) and touch long bone, 1 first appearance right femoral head infarct. Some particular appearances we found. 2 cases of "hand foot syndrome", 1 case ischemic aseptic arthritis and 1 case of vertebra "step staircase". Three salmonella pandiaphysite and 1 tuberculosis spondylodiscite were observed. The authors analyse the major radiological appearances of this hemoglobinopathy and recall the interest in knowing this disease even in low endemicity countries. PMID- 10797984 TI - [Contribution of medical imaging to visceral manifestations of sickle cell anemia in the child]. AB - Sickle cell anemia is one of the qualitative hemoglobinopathies which are the most widespread in the world. In a retrospective study on 11 drepanocytary patients (10 SS and 1 SC) observed at the BORDEAUX children's hospital. 3 Processus were identified: Hemolysis: 3 hepatomegaly, 3 splenomegaly, 1 hematochromatosis and 4 pigmentary lithiasis; Thrombosis: 1 infarctus case of spleen, kidney, lung, 1 papillary necrosis and 1 brain vascular ischemic appearance; INFECTION: 1 spleen abscess and 1 pneumopathy. In 15% of instances, visceral appearances are allowed to discoverer sickle cell anemia. Hemolysis type appearances were currently observed (63.6%) like pigmentary lithiasis and hepatomegaly. The authors analyze the major radiological appearances of this hemoglobinopathy and recall the interest in knowing this disease even in low endemicity countries. PMID- 10797986 TI - [Inhibition of thrombus formation by anti-Willebrand monoclonal antibodies in the guinea pig]. AB - To experimentally evaluate the role of von Willebrand factor in thrombosis, a monoclonal antibody-712-, directed against the binding domain of von Willebrand factor to the platelet glycoprotein Ib and a control antibody D4H1, directed against the light chain of Factor VII C, were injected into guinea pigs. A model of experimental thrombosis induced by laser in guinea pig mesenteric arteries was used. The antibody 712 induced a dose-dependent reduction of the thrombus formation without affecting the bleeding time and the platelet number. Ex vivo, the ristocetin-induced platelet aggregation was inhibited by the antibody 712 in a dose-dependent manner while the adenosine diphosphate-induced platelet aggregation was not modified. In conclusion, this study showed that antibody-712- has a significant antithrombotic activity without inducing a hemorrhagic state, suggesting that the von Willebrand factor--glycoprotein Ib axis was a promising target for the strategy of a new antithrombotic therapy. PMID- 10797987 TI - [Pott's spinal cord compression in the child]. AB - Spinal tuberculosis is the first etiology of spinal cord compression in our areas. We are studying the epidemiological, clinical and therapeutic aspects of Pott's disease. We have collected all children aged from 0 to 15 years admitted for Pott's disease in department of Neurology in CHU Fann SenegaIL from January 1st 1983 to December 31st 1997 spinal tuberculosis. The diagnosis was both clinical and paraclinical. 27 children aged to 2-15 years were hospitalized for Pott's disease. The sex-ratio was equal to 2 in favour of boys. 77.77% of patients were consulted more than a month after the outbreak of the paraplegia. The notion of tuberculous contagion was found in 25.92% of the cases. The main clinical signs were paralysis of the limbs, sensory symptoms and bladder sphincter dysfunction. In 96.29% anomaly of disc and vertebra were recorded, more often affecting two vertebrae. The myelography, performed in 13 patients objectivized an epidural stop in 92.30% of the cases. There was a favorable evolution in 92% of patients under antituberculous chemotherapy. Spinal tuberculosis is an affection cheating all ages and it remains the first etiology of spinal cord compression in our areas. Therefore there is a necessity of primary safety by fighting against tuberculosis what is still in our areas. PMID- 10797988 TI - [Protein C, protein S and antithrombin III at normal delivery and during abruptio placentae]. AB - Protein C, Protein S and Antithrombin III were screened in one hundred patients admitted for abruptio placentae and one hundred women who delivered normally in Dakar university hospital. We found a reduction of Protein S at normal delivery which is linked to hypercoagulation activity during this process. PC and PS were significantly decreased during abruptio placentae in relation with the disseminated intravascular coagulation which was found in our study. We recommend to include these tests to explore aetiologies of abruptio placentae and to confirm their congenital deficit two months after delivery. PMID- 10797990 TI - [Antibodies specific to Plasmodium falciparum antigens in immune individuals: III. Seasonal course of the response to a major antigen of the asexual blood forms in two sites of different malaria exposure]. AB - Specific IgG1 and IgG3 antibody responses to a major Plasmodium falciparum blood stage antigen i.e. MSP1 have been measured in plasma obtained from immune individuals living in areas with different endemicities and sampled at different periods corresponding to different levels of parasite transmission. The study shows a significant imbalance between IgG1 and IgG3 antibody responses in Dielmo vs. Ndiop, and a differential regulation of IgG1 and IgG3 responses both in subclasses and in titers (low: 1/200, and high: 1/2,000) depending upon intensity of parasite exposure. PMID- 10797989 TI - [HLA-DQ allele typing in a Senegalese population: comparative study of serology and PCR-SSP]. AB - In this study, we compared the serological standard typing method and the DNA genotyping PCR-SSP for the characterization of HLA-DQ alleles in a senegalese population. For this purpose, 120 individuals leaving in Dielmo were typed using the microlymphocytotoxicity assay and the PCR-SSP DQ low Resolution method. A discordance of 42.5% (51/120) was found between these two methods. Thirty % (36/120) of serological typed persons failed to be typed by PCR-SSP method whereas 1% (1/120) assigned by PCR-SSP failed to be characterized by serology. Advantages and limits of these two typing methods and also the genetic background of our study population were valid arguments to comment our findings. The PCR SSP, as suggested by several authors, is reliable, accurate and fast for HLA class II alleles characterization. Nevertheless, it needs, to become an alternative HLA typing method, available primers adapted to genetic background of study population. PMID- 10797991 TI - [Micromethod for identification of enterobacteria]. AB - The aim of this study was to set accurate and reliable methods in the identification of Enterobacteriaceae. In Micro CSB Entero each of a strip with 20 cupules containing dehydrated substrates for biochemical identification of bacterial species. Strips were inoculated with 100 microliters per cupule and incubated. After 18 hours, reagents were added. Baye's theorem was used to validate tests. Reactions from Micro CSB were cleared and easily read. 102 strains of Enterobacteriaceae were identified by MicroCSB--89.3% were correctly identified. 0.9% were identified with other tests 9.8% were incorrectly identified MicroCSB gives many advantages: This method save time, space and is less expensive than the other methods and it gives security for identifying species belonging to the different genera. PMID- 10797992 TI - [Micromethod for the identification of streptococcal, enterococcal and staphylococcal species]. AB - The aim of this study was to set accurate and reliable methods in the identification of streptococcal, enterococcal and staphylococcal species. Micro CSB Strep and Staph system consists each of a strip with cupules containing dehydrated substrates for biochemical identification of bacterial species. Baye's theorem was used to validate tests. Reactions from micromethods were clear and easily read. Identification of 229 strains of streptococci and enterococci was correct for most species with 98.7% species with 99.3% sensitivity. 41 strains of staphylococci were also correctly identified with 85.2% of specificity and 97.68% of sensitivity. PMID- 10797993 TI - [Evaluation of acute renal insufficiency in Dakar]. AB - The acute renal failure (ARF) is a serious complication occurring commonly in gravidic toxemia. The aim of this retrospective study was to evaluate the treatments wed on patients presenting an ARF and hospitalized in the Dakar hemodialysis centers and emergency service of Hospital Principal. These patients should be aged over 15 years at present no history of chronic renal failure. The diagnosis of ARF was based on the presence of a creatinine clearance value < or = 80 ml/mn, a creatinemia > 176.8 mo/l, an azotemia > 9.99 mmol/l. Thirty patients aged form 16 to 56 years were studied, they were 6 males and 24 females. Among them, 22 (73.33%) present anuria, 5 (16.7%) were in oligo anuria, and 3 (10%) had a normal diuresis. An aetiologic treatment was instituted whenever necessary. The other treatments consisted in dietary measures, hydroelectrolytic balance, blood transfusion, diuretic therapy, and hemodialysis. All anuric patients had a diuretic regimen. Ten of them had recovered a diuresis, 16 underwent hemodialysis in a mean time of 4 days. None of latter recovered renal function 5 patients developed Chronic renal failure and 11 died, mainly by infectious complications. Acute renal failure has until now a poor prognosis in our regions because of a certain management delay. This management should include a guide and adequate volume replacement, and eventually diuretic therapy, and dialysis as soon as possible. PMID- 10797994 TI - [Ocular burns at the ophthalmologic clinic of the Central University Hospital of Le Dantec apropos of 28 patients]. AB - Chemical and thermal ocular burns are emergencies in ophthalmology because of the blindness sequels they occur. All patients were included in a prospective study form during 18 months. 28 patients were admitted for ocular burns, representing an incidence of 5.21%. 13 of them had occupational ocular burns, 11 homely ocular burns and 4 public way injuries. 18 had chemical burns and 10 thermal burns, with a total of 44 eyes. The clinical observations was dominated by conjunctival lesions: 90.9%, corneal lesions: 79.5%, and plapebral lesions: 43.2%. 11 of them who had chemical burns, the ocular irrigation was made immediately after the accident, and all of them had antibiotic and cicatrizing treatment. The seriousness of ocular burns, the youngness of patients must instigate the authorities to improve the working conditions in the factories, and a better information in manipulation of dangerous chemical product. PMID- 10797995 TI - [Nutritional state of women and children in the rural community of Mpal (District of Saint Louis, Senegal)]. AB - This cross sectional survey carried in april 1995 aimed to assess the nutritional status of the women old enough to procreate++ and the children less than 5 years old in the community of Mpal (District of St Louis). The method of sampling used was the method of stratified poll. The assessment of the nutritional status is carried among the women by measuring the brachial circumference and the corporal mass index. Among the children, the indicators used were the ratios: weight-age, weight-height and the brachial circumference. The results showed a rate of malnutrition very high in comparison to the national averages as well in ythe women old enough to procreate (31% versus 17.4%) as in the children (11% of acute malnutrition versus 8.7% and 24.4% of chronic malnutrition versus 21.7%). The young mothers and the children more than 3 years old were the more frequently reached. It urges to set up in this locality a programme of struggling against malnutrition based on the integration of the economics activities of the communitary groups to their health activities. PMID- 10797996 TI - [Neuroepidemiology of epilepsy in Senegalese school milieu]. AB - The authors proceeded to a 7 month long prevalence survey on epilepsy in the schools of the regions of Dakar and Thies in Senegal, West africa. It consisted to a cross sectional descriptive study targeting the children between 3 and 10 years old. The W.H.O. questionnaire on epilepsy was used for data collection. Among the 2803 children of the population study, 58 were detected suffering from epilepsy, realizing a prevalence rate of 21%. The males were more represented (62% of the epilepsies). All the epileptic children presented generalized seizures represented either by "Grand Mal" generalized tonic-clonic form (84.4% of the epilepsies), or by "Petit Mal" absences (15.6% of the epilepsies). The etiological forms were dominated by idiopathic epilepsies. Secondary generalized cases represented 31% of the epilepsy and were essentially detected in rural areas. They were usually related to obstetrical traumas (61.1% of the secondary generalized epilepsies) or to child encephalopathy with seizure attacks (38.9% of the secondary generalized epilepsies). It appears also from this study that epilepsy remains a dramatic public health reality and is more frequent among low economy income people who have less access to the health care system. PMID- 10797997 TI - [Survey of nosocomial infection prevalence in the surgery department of the Central National Hospital of Ouagadougou]. AB - The objective of this study is to collect preliminary epidemiologic data about nosocomial infections (NI) in the National Hospital of Ouagadougou. A prevalence survey of NI was realized in surgical wards (116 patients). All hospitalized patients were included, excepted those stayed less than 48 hours. Using the definitions of the "Conseil Superieur d'Hygiene Publique de France", the nosocomial infections studied were surgical wards, pneumonia and urinary tract infection. The result showed that 19 patients (16.4%) had 26 nosocomial infections (prevalence: 22.4%). Six patients had at least 2 nosocomial infections. The infection rate increases with operation, debilated preoperative physical statute, and invasive care. The average increasing of duration of hospital stay for infected patients was 10 days. This data confirm the importance of nosocomial infections and emphasize the need of surveillance and control of nosocomial infections. But deeper epidemiological studies must be undertaken before. PMID- 10797998 TI - [Importance of the number of treatment sessions in the success of root canal therapy]. AB - Root canal therapy is usually performed in single or multiple-visit treatment. The choice of single or multiple visit depend on dentist and patient available time. In general dentistry, multiple-visit is widely chosen by dentist whereas endodontic speciality recommend single-visit. In this study we analyzed 120 root canal therapy performed by students, to determine the number of visits needed to achieve treatment from endodontic-cavity-access to root canal filling and its possible, influence in the successful rate. Results showed that globally, 60% of treatments were performed in multiple-visit. Results showed also hat 89.3% of anteriors teeth were performed in single-visit but there is a higher rate of fail: 31.78%. Nevertheless, these results do not condemn endodontic single visit therapy. PMID- 10797999 TI - [Incidence of postoperative pain in single session root canal therapy (study in black senegalese apropos of 96 cases)]. AB - This prospective study was conducted during a period of 30 days in 96 patients to determine the incidence of postobturation pain after single-visit root canal treatment in black Senegalese people. The canals of all teeth were hand-prepared and filled with zinc-oxide-eugenol paste by a Lentulo spiral paste filler. Patients were asked to categorize their pain according to the following criteria: No pain, Slight pain, Moderate pain and Severe pain. The data were analysed statically with chi-square test to determine the relationship, if any, between the pain experienced and sex, age, tooth type, apical level of root filling and number of treatment performed in the single-visit. Results show that eighteen patients experienced pain (18.75%); our results are ranging between 10% and 25% described in the literature. No significant correlation was found between postobturation pain and any other factor with the exception of the apical length of canal filling. Characteristics of postobturation pain after root canal treatment performed in single-visit in Senegalese people presents the same features as described in international papers, that is, pain appears early and analgesic therapy is effective. PMID- 10798000 TI - [Acute carbamate poisoning: apropos of 7 cases occurring in a Senegalese industrial unit]. AB - The authors describe an onset of acute pesticide intoxication which occurred in an industrial site in Dakar. The substance responsible belongs to the class of carbamates which leads to variety of symptoms in relation with acetylcholin accumulation. All seven employees involved were young, males, unqualified temporary workers. Many factors appear to facilitate the intoxication: heat stress, inappropriate protective devices, and lack of information on pesticides' hazards. Routes of entry were cutaneous and respiratory. Symptoms were related to muscarinic and nicotinic syndromes. The course of this episode was favourable because of adequate treatment. PMID- 10798001 TI - [Hepatic tuberculosis of the pseudotumoral form]. AB - The authors report a case of a 30 years old immunocompetent woman with liver tuberculosis with an unusual pseudotumoral presentation and secondary occurrence of abcedation with cutaneous fistulization. The diagnosis was based on the bacteriological positivity for acido-alcoolo resistant bacillus in pus of the abscess obtained by ponction guided by ultrasonographic examination. The authors emphasize in the differential diagnosis with the other causes of liver abscess (amibiasis and pyogenic microorganisms) and liver carcinoma. They also note the importance of the function guided by ultrasonographic examination permitting histologic and bacte+eriologic study in the diagnosis of this unusual presentation of liver tuberculosis. The patient was treated by antibacillary antibiotics and evacuation function. The prognostic was good with 6 months of follow-up. PMID- 10798002 TI - [Fibrodysplasia ossificans progressiva or Munchmeyer disease apropos of 2 cases]. AB - Fibrodysplasia ossificans progressiva or myositis ossificans progessiva or still Munchmeyer disease is a genetic ailment with dominant autosomic transmission. It includes a high rate of change and doesn't appear any race. The authors gave an account of two remarks about it on a causal and late discovery upon two young black Africans. The first observation is a nine year-old-boy, without any similar family previous history and who has been hospitalized in maxillo-facial milieu for a mandible osteitis staphiloccocus. It showed some muscular ossifications of paravertebral, cervical, dorsal and lumbar nature--from a spontaneous appearance and evolving progressively since the age of three months. The radiographic results displayed some specific bones disorders which enable to retain the diagnosis above. The second observation is the case of a twenty four-year-old woman who has previously benefitted from a surgical exploration a non inflammatory muscular tumefication at the right arm which occurred at the age of twenty one. Three years later, she took surgery for the restriction of the oral gap and a right hand side lumbar paravertebral tumefaction which was thus restricting the mobility of the rachis. The radiological results have found some specific bone disorder++ at the hands and the feet which enable to link the paravertebral ossifications to their fibrodysplasic origin. Munchmeyer disease remains a affection of easy radio-clinical diagnosis. The essential point is to precociously think of it before the specific osseous anomalies occurring at the level of the feet and the hands. The functional or even vital prognosis remains closely linked to both the important and the topography of conjunctivo-muscular ossifications. PMID- 10798003 TI - [Apropos of 1 attempted suicide with a fire arm in an adolescent]. AB - This is the presentation of an adolescent's clinical case who tried to commit suicide with a fire arm during a depressive crisis. An emphasis is placed on the family interaction model and on a more global approach of the patient's personality. The authors underlined some of the problems raised by the diagnostic approach linked to the delirious syndrome, full of psychotic elements: themes of persecution, mystico-religious themes associated with impulsive escapade, all of them going with a depressive state. They gave also an important place to a pluridisciplanary treatment in those particular stages of crisis. PMID- 10798004 TI - [Biermer's disease without anemia: apropos of 2 cases manifested as glossitis with macrocytosis]. AB - Although it is recognized that glossitis is a classical sign of pernicious anemia, occurring in the evolution of this disease, it is unfrequent for this sign to reveal this affection. We report two cases where diagnosis was evacuated on the presence of glossitis and macrocytosis despite absence of anemia. Confirmation was done by low serum cobalamin level, gastritis atrophy and presence of intrinsic factor antibody. We emphasize that increased clinical suspicion may lead to early diagnosis even if anemia is lacking. PMID- 10798005 TI - [Neuroendocrine carcinoma: first case observed in Dakar]. AB - Neuro endocrine carcinoma is an unusual malignant tumor occurring in white elderly persons. It's most common location is the skin of head and neck. We report the first case of of Merkel cell carcinoma from Dakar. It was a woman of 21 years with history of three nodular lesion on the left foot which occurred 8 month before her arrival in our department. Inguinal and poplital lymph nodes were enlarged. Histopathology and immunocytochemistry confirmed the diagnosis of Merkel cell carcinoma. Our case is remarkable by the young age of our patient, her black race, the relative importance of the size and the localization of the tumor and the early occurrence of lymph node and pulmonary metastases. PMID- 10798006 TI - [First case of primary IgA glomerulonephritis (Berger's disease) in Senegal]. AB - Berger's disease or IgA glomerulonephritis is the most common glomerular nephropathy in Europe and represent a rare event in blacks. Here, we describe the case of a 43 years old black Senegalese whose disease was discovered while investigating a persistent proteinuria with high blood pressure and chronic renal failure, but without hematuria. We point out the uncommon feature of this clinical presentation and the importance of bad prognostic factors presented by this patient. We obtained a good outcome by means of converting enzyme inhibitors and corticosteroid therapies: regression of renal failure and normalization of blood pressure. The generalization of renal biopsy practice would lead to a better knowledge of the incidence of this disease among Senegalese people. Indeed, renal biopsy is the main tool to diagnose glomerulonephritis and subsequently adapt the therapy aimed at preventing the possible evolution to end stage renal disease. PMID- 10798007 TI - [Tracheal atresia: apropos of 2 cases]. AB - Two cases of tracheal atresia are reported. This is a rare malformation which is responsible of neonatal respiratory distress, lack of cry at born and difficulties to put a tube inside the trachea. Various malformative abnormalities are commonly associated to tracheal atresia. The death occurred in these two newborns some days after birth. Tracheal atresia is a common lethal malformation. PMID- 10798008 TI - Subtypes of codeine cough syrup abusers. AB - This paper highlights the abuse potential of the codeine containing cough syrups, which may take two forms. One, experimental abuse in school or college students which later persists in a dependent pattern. Two, pre-existing opioid abusers, as a substitute which starts after the school or college years. The short term treatment outcome is better in the former group in that they are able to maintain abstinence for a relatively longer period. PMID- 10798009 TI - Clinical significance and evaluation of proteinuria in NIDDM patients. AB - The proteinuria may reach massive proportions resulting in the nephrotic syndrome. Early and simple diagnostic may prevent further complications. Attempts are made to correlate a simple parameters like pH, specific gravity, excretion of reducing sugar and protein profile on SDS-PAGE, an advanced technique. The pH ranged between 5.7 to 9. However, specific gravity of the urine showed significant increase from 1.000 to 1.088 and has linear relation with percentage of sugar (0.2 to 2 gm) and protein banding 1 to 5 on SDS-PAGE. PMID- 10798010 TI - Acute appendicitis--an unusual cause. PMID- 10798011 TI - Streptogramins: a new class of antibiotics. AB - Streptogramin antibiotics represent a unique class of antibacterials in the each member of the class consists of at least 2 structurally unrelated molecules: group a streptogramins (macrolactones) and group B streptogramins (cyclic hexadepsipeptides). Both group A and group B streptogramins inhibit protein synthesis at the ribosomal level, and they act synergistically against many isolates their combination generating bactericidal activities and reducing the possibility of emergencies of resistant strains. The mechanisms of acquired resistance to group B streptogramins remain unaffected by target modifications and active efflux. The pharmacokinetic parameters of group A and group B streptogramins in blood are quite similar. In addition, both the A and B group penetrate and accumulate in macrophages and in the bacterial gegetations of experimental endocarditis. Until recently, the complex and irregular composition of naturally occurring pristinamycin and virginiamycin, as well as the unavailability of soluble forms, have limited the clinical development of streptogramins. The synthesis of water soluble derivatives of pristinamycin IA and IIB has now allowed the development of injectable streptogramins with fixed compositions. This unique class of antibacterials will have a significant clinical impact in a world of increasing multidrug resistance affecting the Gram positive cocci, especially staphylococci and pneumococci. The absence of cross resistance to macrolides in many of these isolates and the rapid antibacterial killing against these species bright future for this class of antibiotics. PMID- 10798012 TI - Breast self examination for early detection of breast cancer. AB - Breast cancer is the commonest form of cancer in women in Western countries and second most common in women of developing countries like India. In the absence of an exact aetiological agent for breast cancer, the most appropriate way of controlling it is by early detection and treatment. Of the various methods of screening for breast cancer, mammography is the method of choice but its use is limited due to high cost and unavailability. Considering this, breast self examination (BSE) is an ideal method which can be done by every woman at her leisure time with little training. Medical and paramedical professionals can act as trend setters in promoting BSE for control of breast cancer in the community. PMID- 10798013 TI - Psychogenic cough--a case report. AB - A case study of a 12 year old child evidencing psychogenic cough is presented. The importance of behavioral and family factors in maintaining such cough and an integrated psycho-behavioral approach at management are described. A 1 year follow-up revealed a rapid and complete elimination of psychogenic cough and good adjustment. PMID- 10798014 TI - FDA clears tuberculosis drug for marketing. PMID- 10798015 TI - The Krever recommendations: compensation, an integrated blood service and revamped watchdog. PMID- 10798016 TI - Comparative values of CSF-LDH isoenzymes in neurological disorders. AB - The present study was carried out to evaluate the usefulness of Cerebrospinal fluid (CSF) Lactate dehydrogenase (LDH) isoenzymes in the diagnosis in tuberculous meningitis (TBM), pyogenic meningitis (PM), viral encephalitis (VE) and hydrocephalus (HC). A characteristic dominance of isoenzymes in cerebrospinal fluid was observed: LDH4 in TBM while LDH3 in PM. However, in VE and HC, LDH2 and LDH1 were dominant respectively. The control subjects revealed the presence of isoenzymes LDH1 and LDH2 in very low concentrations. Pattern of LDH isoenzymes in CSF may serve as a diagnostic tool to differentiate these neurological disorders. PMID- 10798018 TI - Predominant phage types of coagulase positive staphylococci in hospital infections. AB - 360 strains of Staphylococcus aureus isolated from various clinical specimens were subjected to bacteriophage typing. 247(68.6%) strains were typable. Among the typable strains 75(20.83%) belonged to phage group I, 45(12.5%) belonged to phage group III, 6(1.67%) belonged to phage group II and 14(3.89%) strains belonged to miscellaneous group. By far, the largest was the mixed group having 107(29.72%) strains. 113 strains (31.4%) were untypable. All the strains were tested for antibiotic sensitivity test. 287 (79.7%) were multiple drug resistant strains. PMID- 10798017 TI - Salmonella typhi VI antigen co-agglutination test for the rapid diagnosis of typhoid fever. AB - A slide Co-agglutination test for the detection of Salmonella typhi Vi antigen in blood was evaluated for its efficiency in rapid diagnosis of Typhoid fever. The results were compared with conventional methods like Blood culture and Widal test. The test showed a sensitivity of 86.67% and specificity of 88.83% when compared with blood culture positivity or Widal titre above 160. This is a useful rapid diagnostic test for the early diagnosis of Typhoid fever. PMID- 10798019 TI - Teenage pregnancy outcome: a record based study. AB - Present record based study was undertaken in Medical record section of Government medical college, Nagpur, to assess teenage as a risk factor for pregnancy complications, outcome, and operative or assisted delivery. Five year (January 1993 to December 1997) data was scanned, which gave sample of 1830 teenage pregnancies; while equal number of subsequent partly matched controls (> 20.29 years) were taken. Results showed proportion of low birth weight baby to be significantly greater in teenagers (p < 0.001). Operative interference was significantly greater in adult pregnancies (p < 0.001). Though stillbirth and preterm delivery was more in cases, difference was not statistically significant. Similarly toxaemia of pregnancy, premature rupture of membrane, placenta previa, accidental haemorrhage though more in adult pregnancies was statistically not significant. There were no differences in cogenital anamoly and twins between cases and controls. But breech deliveries were significantly (p < 0.001) more in adults. PMID- 10798020 TI - Blood transfusion practices--a case study. AB - Blood transfusion practices of the treating doctors in a district hospital in Haryana were studied through retrospective study of blood bank records for the years 1992 to 1994 and interview of the clinicians of various specialities in the hospital. It was found that utilization of the whole blood was 90%, 89% and 81% respectively of the total blood units utilized during this period. Single unit transfusions out of the total transfusions done were 87% in 1992 and 1993 while these were 89.9% in 1994. Blood was often requested for volume replacement in acute haemorrhage. Only homologous blood transfusions were done at the hospital. This study has highlighted that there is scope for improvement of blood transfusion practices by strictly following the indications for use of blood, promoting the preparation and use of blood components, use of plasma expanders for acute blood loss, avoiding single unit transfusions and promoting the use of autologous blood during routine surgery. PMID- 10798021 TI - Foetal medicines. PMID- 10798022 TI - Effects of flunarizine on dopamine dependent behaviours in rats. AB - 1. Radio-ligand binding study has demonstrated that flunarizine has a high affinity for the rat striatal D 2 dopamine (DA) receptors. 2. In the present behavioural study conducted in rats it was observed that flunarizine, unlike the postsynaptic striatal D 2 DA receptor agonist apomorphine, did not induce stereotyped behaviour (SB) in rats. This indicates that flunarizine does not act as an agonist at the postsynaptic striatal D 2 DA receptors. 3. Flunarizine however, like the postsynaptic striatal D 2 DA receptor antagonist haloperiodal, inhibited the conditioned avoidance response, induced catalepsy and antagonized the SB induced by the DA agonists apomorphine and methamphetamine. 4. Our findings indicate that flunarizine acts as a postsynaptic striatal D 2 DA receptor antagonist. PMID- 10798023 TI - Primary amenorrhea with Xq duplication. AB - Two female patients aged 16 and 17 years with Turner features short stature, amenorrhea and gonadal dysgenesis were referred for cytogenetic confirmation and counselling. Their karyotypes were 46, X, dup (X) (q13-->q22). The clinical and the cytogenetic picture have been correlated. PMID- 10798024 TI - Sion Hospital--a historical sketch (inception--1947 to beginning of Sion Medical College--1964). PMID- 10798025 TI - New observations on carrying angle. AB - Based on experiments on fresh cadaveric and accidentally amputated 8 upper limbs of children, study of ulnae for presence and absence of non articular strip on the trochlear notch, measurements of carrying angle, length of forearm bones, pronation-supination, height and weight in 2250 infants, children and adults of various age groups and clinical observations on 800 cases of injuries around elbow many new facts have been observed about the development of the carrying angle and its significance in the etiopathogenesis of various types of fractures seen around the elbow. The carrying angle develops in response to pronation of the forearm and is dependent on length of the forearm bones. Lesser the length of forearm bones greater is the carrying angle. So the carrying angle is more in shorter persons as compared to taller persons. It is abduction at the shoulder and not the carrying angle which keeps the swinging upper limbs away from the side of the pelvis during walking. Carrying angle is not a secondary sex character. The type of fracture a child sustains after fall on outstretched hand is also determined by the value of the carrying angle. A new type of fracture hitherto undescribed in the literature, T-Y fracture of the distal humeral epiphysis is also reported. PMID- 10798026 TI - Prediction of birth weight from simple anthropometric measurements in Indian male and female new born babies. AB - An attempt has been made in this paper to select suitable combination of predictors of birth weight from three anthropometric measurements (viz. length, head and chest circumferences) taken at birth of Indian male and female new born babies. The best combination of predictors was selected by examining the separate effects of each of the three concomitant variables on birth weight using the technique of multivariate regression analysis. The combination of head and chest circumferences was found to be the most appropriate for estimation of birth weight in view of the simplicity and non-invasiveness of measuring these two body circumferences. Keeping in view of the high multiple correlations, nomograms have been constructed to predict birth weights from head and chest circumferences in male and female new born babies. These nomograms will serve well as a simple and quick methods for predicting birth weight from head and chest circumferences in Indian male and female new born babies whenever the Direct weighing at birth is not feasible. PMID- 10798027 TI - Munchausen's syndrome: a case report. AB - Munchausen syndrome, a chronic factitious disorder with physical symptoms, also called hospital hoboes or hospital addiction, is an uncommon psychiatric disorder, usually presents as an emergency. The correct recognition of the syndrome is important in medical practice. A brief review with an interesting case report is presented. PMID- 10798028 TI - Sudden cardiac death in the young athlete. AB - Sudden cardiac death in athletes is usually due to underlying cardiovascular disease. In the young less than 30 years of age, the most common abnormality is hypertrophic cardiomyopathy, followed by congenital coronary artery anomalies. The final common pathway is usually ventricular fibrillation. Sudden cardiac death in the young is rare but remains a source of concern. A careful screening history and physical examination, especially for potential athletes, should identify the majority of young people at risk. PMID- 10798029 TI - Effect of handwashing agents on bacterial contamination. AB - In order to assess the effect of handwashing agents like soap and ash on the control of bacterial contamination, the study was carried out in two villages of Rajendranagar mandal of Rangareddy district, Andhra Pradesh. Twenty households belonging to high income group and twenty households belonging to low income group having 1-2 years age children were randomly selected for the study handwash samples. Before feeding the child handwash samples after washing with different agents were collected and analysed for bacterial contamination. The study revealed that use of soap or ash for washing hands before feeding the child reduced hand contamination significantly. PMID- 10798031 TI - Neonatal morbidity in a hospital at Shimla. AB - During the study period there were 2063 live births. Of these 573 (27.8%) were low birth weight (LBW), 277 (13.4%) preterm and 148 (7.1%) small for date (SFD) babies. In all, 263 (12.7%) newborns suffered from one or the other morbidity. Birth asphyxia of varying severity developed in 130 (6.3%) babies [88 LBW and 42 normal birth weight (NBW) (p < 0.001)]. Respiratory distress syndrome was diagnosed in 82 (3.9%) babies, most being due to hyaline membrane diseases (31.7%), which affected 26 (9.4%) of preterm babies. Deep infections were seen in 109 (5.3%) newborns [60 LBW and 49 NBW, (p < 0.001)] and superficial infections were seen in 79 (3.8%) babies [46 LBW and 33 NBW, (p < 0.001)]. Hyperbilirubinemia was detected in 78 (3.8%) babies. In one fifth of the babies, the cause of hyperbilirubinemia remained unidentified even after detailed investigations. Hypothermia was observed in 59 (2.9%) newborns [48 LBW and 11 NBW, (p < 0.001] and congenital malformations were seen in 24 (1.7%) babies. Morbidity was found to be high amongst LBW and preterm babies. The incidence of deep infections and hypothermia was high in our study. PMID- 10798030 TI - Microbial profile of neonatal infection in Coimbatore. AB - In this study, 187 consecutive neonates suspected of having septicaemia were investigated for isolation of micro organisms. Two samples of blood were collected for isolation of aerobes and anaerobes. Cultures were positive in 75 (40%) cases. Aerobic bacteria were the major etiological agent, accounting for 93% of positives including the 8% cases showing polymicrobial etiology. Anaerobic bacteria and Candida species were isolated in 6.6% and 8% of positive cases respectively. Bacteroides fragilis (amongst anaerobic) and Staphylococcus aureus (amongst aerobic) were the predominant organisms isolated. Clinical presentations were not specifically different to distinguish aerobic from anaerobic bacteria. In the present study, 6.6% of bacteremias were due to anaerobes, hence possibility of some of the bactermias being due to anaerobes should be kept in mind while treating cases of neonatal septicaemia. For a complete microbial profile both aerobic and anaerobic cultures should be done. PMID- 10798032 TI - Role of protozoa as risk factors for persistent diarrhea. AB - A case control study including 175 children aged 0-36 months suffering from diarrhea of > or = 14 days duration was undertaken to determine whether there is an association between Giardia lamblia, Entamoeba histolytica or Cryptosporidium infection and persistent diarrhea (PD). Subjects were identified by ongoing household surveillance and enrolled as cases. For each case two controls were selected by survey of neighbouring households--a child with acute diarrhea and one without diarrhea. Both the controls were matched with the case for age and nutritional status. Two fresh stool samples were collected from all cases and controls at enrollment and examined for trophozoites of Giardia lamblia, Entamoeba histolytica and Cryptosporidium. Giardia lamblia trophozoites were detected in a significantly higher proportion of PD cases (20.0%) than acute diarrheal and non diarrheal controls (4.6% each, p < 0.0001). There were no significant differences in the proportion of cases and controls who passed E. histolytica trophozoites or cryptosporidium in their stools. There was a consistent trend towards poorer weight gain in PD cases who passed Giardia trophozoites in stool; the differences were statistically significant at days 14 and 21, after enrollment. Giardia lamblia infection is more prevalent in PD cases than in acute diarrhea or non-diarrheal controls. This prevalence is not high enough to warrant routine anti-giardia therapy in patients with PD. However, as giardiasis was observed to have adverse growth impact in PD cases, stool microscopy for detection and subsequent treatment of Giardia lamblia seems to be justified. PMID- 10798033 TI - Lead exposure and iron deficiency among Jammu and New Delhi children. AB - In order to examine the prevalence of lead exposure and iron deficiency in Jammu, blood lead (B-Pb) and erythrocyte protoporphyrin (EP-ZnPP) levels were measured in a group of 125 children and compared with B-Pb and EP levels of 46 New Delhi children. The mean B-Pb level of Jammu children was 15 micrograms/dl and ZnPP level 46 micrograms/dl. The frequency distribution of B-Pb in Jammu children according to the 1997 CDC stratification showed that only 33% were below the 10 micrograms/dl "normal" threshold levels and 50.5% were between 10-19 micrograms/dl, an increasing level of concern. The remaining 16.5% were in the medical intervention lead poisoned level between 20-87 micrograms/dl with corresponding EP levels between 29-160 micrograms/dl. The mean B-Pb and ZnPP levels for New Delhi children were 14 micrograms/dl and 55 micrograms/dl respectively. This study on Jammu children shows a modest increase in mean B-Pb and EP levels from those in 1985; the percentage of children with higher levels of B-Pb > 10 micrograms/dl have significantly increased. Moreover, the continued underlying high prevalence of iron deficiency among these children predisposes them to increased lead absorption from various sources thus aggravating further, the detrimental effects of lead. Therefore, a more extensive screening of all children below 10 years of age and an ongoing B-Pb and EP monitoring and iron supplementation programme for Jammu and New Delhi is essential. An environmental assessment of the sources leading to serious lead intoxication problem in Jammu, a rapidly growing and overpopulated city of the state of Jammu and Kashmir is strongly recommended. PMID- 10798034 TI - Vitamin D deficiency rickets at birth in Kuwait. AB - In the present study vitamin D deficiency rickets has been diagnosed within 24 hrs. of birth. Seventy five full term, otherwise healthy newborns, weighing more than 2.5 kg were born with rachitic rosary. 25-Hydroxyvitamin D was lower than normal in 56 newborns and 15 mothers. Alkaline phosphatase was higher than normal in 26 and radiological changes suggestive of rickets were seen in the wrist X ray of only 14 newborns. Hyperphosphataemia was present in all the newborns. 1,25 dihydroxyvitamin D was markedly increased in six out of ten newborns. PMID- 10798035 TI - Effect of passively transferred anti-poliovirus antibodies on seroconversion. AB - A prospective study enrolling 50 mother-infant pairs was undertaken to determine the effect of maternal antibodies on poliovirus antibody titres and seroconversion rates in infants and to determine the difference in titres and seroconversion rates following three and five doses of oral poliovaccine (OPV). Cord blood samples and samples collected 4 weeks after 3rd and 5th doses of trivalent oral poliovaccine were processed for estimation of anti-poliovirus antibody titres. These were expressed as geometric mean titres (GMT). Significance was analyzed using unpaired 't' test. The relationship between maternal antibody titres and seroconversion was determined by correlation coefficient test. Post OPV5 titres were significantly higher than post OPV3 titres for type 1 and type 2 polioviruses. Seroconversion rates against type 1, 2 and 3 polioviruses were 92.9%, 100.0% and 92.9% following OPV3 and 100.0%, 100.0% and 93.2% following OPV5. The cord blood titres did not have any relation to post OPV3 or post-OPV5 titres. Although there is significant passive transfer of poliovirus antibodies across the placenta, this does not affect titres achieved after immunization. Post-OPV5 titres against type 1 and type 2 viruses are significantly higher than post-OPV3 titres. The seroconversion rates following OPV5 are higher than those obtained post-OPV3 but this difference is not statistically significant. PMID- 10798036 TI - Urinary continence following posterior urethral valves treatment. AB - This retrospective study discusses the incidence of symptomatic voiding dysfunction, the etiological factors and the changes with age and with increasing duration of follow up in 67 boys treated for posterior urethral valves and followed up for a period ranging from 1-21 years. Twenty three of the 67 patients (34.3%) had symptomatic voiding dysfunction; the commonest symptom being nocturnal enuresis with diurnal urgency and frequency. Data in this study suggests a definite decrease in the incidence of urinary incontinence as the patient grows older or as the duration of follow up after valve fulguration increases. A Tc-99m DRCG based uroflowmetry was available in 31 of the 67 patients at the time of this review and evidence of detrusor dysfunction was identified in 9 of the 31 patients on the basis of uroflowmetry. Of significance was the detection of detrusor dysfunction on uroflowmetry in 3 patients with a history of normal urinary control and urinary stream with persistent post treatment hydroureteronephrosis eventually culminating into renal failure. Imipramine therapy was given to 10 of the 17 patients with symptoms of urinary frequency, urgency and nocturnal enuresis with a significantly symptomatic improvement in 9 of these patients. Urinary incontinence, detrusor dysfunction and its effect on the upper tracts and renal function is a significant determinant of the long term outcome of the boys treated for posterior urethral valves and this paper emphasises on the need to carefully evaluate and treat this aspect of all valve patients. PMID- 10798037 TI - Lessons learnt from Diarrheal Diseases Control Program and implications for the future. AB - The national Diarrheal Disease Control Program was launched with the aim of improving the knowledge and practice of appropriate case management among caretakers of young children as well as health care providers. The National Family Health Survey (1992-3) revealed that 42.7% of mothers knew about ORS packets and 25.9% had ever used them. ORS use rates in children who suffered from diarrhea during the previous two weeks varied from 8.3% in Rajasthan to 50.1% in West Bengal. These findings represent substantial accomplishment, and also are a reminder that we still have to reach more than half of the households. Further analysis of the NFHS data showed that exposure to electronic mass media had a significant impact on mothers' awareness about ORS packets (56% in exposed, 32% in unexposed) and ORT use rates (38% in exposed, 25% in unexposed). In this review, future strategies for increasing the impact of the program are discussed. These include involvement of licensed and unlicensed medical practitioners, greater use of the electronic mass media, ORS standardization, greater focus on poorly performing states and resolving residual issues in the case management of diarrhea in malnourished children and infants under 6 months of age. PMID- 10798038 TI - Kala-azar--new developments in diagnosis and treatment. AB - Kala-azar is an endemic disease in many parts of India. Traditionally, diagnosis of this disease was based on demonstrating the parasites in various tissues like bone marrow or splenic aspirates. However, lack of high sensitivity of these methods led to the use of various immunodiagnostic methods in the diagnosis of kala-azar. Antigen detection and polymerase chain reaction to detect parasitic DNA have been found to be useful in patients with an underlying immunosuppressive disease like AIDS. For treating kala-azar, pentavalent antimonial compounds are still the first-line agents. However, due to increasing resistance to this agent, many patients at present require other drugs including amphotericin B and pentamidine. Toxic effects of these second-line agents have led to development of drug delivery systems like liposomal amphotericin B, which has shown uniform efficacy in clinical trials. Combining stibogluconate with either paromomycin or interferon-gamma has also been shown to be useful in many patients with drug resistant kala-azar. PMID- 10798040 TI - Prevalence of intestinal pathogens in HIV patients with diarrhea: implications for treatment. AB - Patients infected with the human immunodeficiency virus (HIV) commonly experience diarrhea at some time during their illness. A variety of enteric pathogens are identified in 50-80% of these patients, depending on the intensity of the diagnostic work-up that is done. In addition to the common enteric pathogens, several unusual enteric pathogens are recognized to cause diarrhea especially in HIV patients. These include protozoan parasites such as Cryptosporidia, Isospora belli, Cyclospora cayatenensis and Microsporidium species bacteria such as enteropathogenic Escherichia coli and Mycobacterium avium-intracellulare, fungi including Candida albicans and Histoplasma capsulatum, and viruses such as astroviruses and caliciviruses. Diagnosis of these infections sometimes involves special procedures not readily available every where, and empiric therapy based on knowledge of the likely pathogens has been advocated for developing countries. This article reviews the currently available data on geographic variation of enteric pathogens in HIV patients with diarrhea and outlines a rational strategy for empiric therapy of these patients. PMID- 10798039 TI - Viral encephalitis of public health significance in India: current status. AB - Japanese encephalitis (JE) and rabies are 2 viral encephalitis that are of public health importance in India. JE is a zoonosis with the primary cycle occurring in arthropods (mosquito vectors) and vertebrate animals (primarily the pig), man being only an incidental 'dead end' host. Out-breaks have been seen in most parts of India except the north west. The disease presents with a prodromal stage, an acute encephalitic stage with coma, convulsions and variable deficits and a convalescent stage. Diagnosis can be made by viral isolation from CSF or brain, or serologic tests such as haemagglutination inhibition test and IgM antibody capture ELISA in CSF and blood. There is no specific treatment. Mortality ranges from 20-50% and almost half the survivors have sequelae. The most effective control measure besides control of mosquitos is vaccination. A killed mouse brain vaccine is being prepared in India and is safe and effective but expensive. Rabies is a highly fatal encephalomyelitis primarily occurring in urban dogs and wild animals especially canines. It is endemic in India and affects an estimated 3 per 100,000 persons annually. The patient initially may display bizarre combative behaviour. The disease can be effectively prevented by post exposure vaccination. The nervous tissue vaccine is no longer recommended because of unacceptable neurotoxicity. Three cell culture vaccines are presently available with about equal efficacy. PMID- 10798041 TI - Dengue hemorrhagic fever: clinical manifestations and management. AB - Dengue virus infection may remain asymptomatic or manifest as nonspecific viral infection to life threatening dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS). Patients with DHF/DSS have fever, hemorrhagic manifestations along with thrombocytopenia and hemoconcentration. Thrombocytopenia and hemoconcentration are distinguishing features between DHF/DSS and dengue fever (DF). Some patients with dengue fever may have significant bleed and mild thrombocytopenia but no hemoconcentration. These patients are labelled to have dengue fever with unusual bleeds. Laboratory findings in DHF/DSS include rising hematocrit, thrombocytopenia and transformed lymphocytes on peripheral smear. There may be increased transaminases, hyponatremia, transient increase in blood urea nitrogen and creatinine. In severe disease there may be lab evidence of dissemination intravascular coagulation. X-ray film of the chest may show pleural effusion. Ultrasonogram of abdomen may detect thickened gall bladder wall with hepatomegaly and ascitis. In some patients there may be abnormality in electrocardiogram and echocardiogram. The diagnosis of DHF/DSS is based on typical clinical findings. For confirmation of dengue virus infection viral culture can be done on blood obtained from patients during early phase of illness. In later part of illness antibodies against dengue virus can be demonstrated by various techniques. The treatment of DF is symptomatic. For control of fever nonsteroidal anti-inflammatory drugs should be avoided. DHF/DSS are managed by intravenous fluid infusion with repeated monitoring of vital parameters and packed cell volume (PCV). PMID- 10798042 TI - Recent developments in the management of malaria. AB - The last decade has seen in increase in our understanding of the pathophysiology of severe malaria in both adults and children. However, this increased knowledge has yet to be matched by progress in the clinical management of this medical emergency. In the last few years a wide variety of measures have been advocated to lessen the mortality due to severe malaria, but none have shown a significant benefit in terms of mortality or morbidity. In this review we concentrate on aspects of the pathophysiology of malaria which are amenable to intervention at present or in the near future. In the field of uncomplicated malaria problems remain mostly related to the continued spread of drug-resistance and the limited array of available drugs. Outlined are a number of important advances in antimalarial pharmacology and parasite biology that may lead to future improvement in the care of patients with malaria. PMID- 10798043 TI - Considerations of HAV vaccine in India. AB - Currently available evidence indicates that most parts of India still quality to be hyper-endemic regions and hence recommendations for vaccinations developed for low endemicity regions are not applicable. There are however, some pockets which showed evidence of distinct epidemiologic shift. There is need to identify geographic regions like Kerala which have potential for epidemic outbreaks through cyclic sero-epidemiological surveys. With economic development and consequent improvements in the levels of sanitation and quality of water supplies, more such areas will be identified. Efforts to improve sanitation and personal hygiene will remain as the most important and efficient intervention to retard the transmission of HAV. Unless a critical improvement in the living standards of our population is achieved, aim of eradicating HAV infection from the community is not realistic. Presently, the cost of three doses of HAV is exorbitant and the focus of HAV vaccination will have to be restricted to individual protection who are likely to remain unexposed till adulthood and can afford to pay for it. HAV Infection below 5 years is mostly asymptomatic. The most efficient use of resources will therefore be to offer HAV vaccine to high risk individuals beyond this age after screening for antibodies against HAV. PMID- 10798044 TI - Pain in neonate. AB - Anatomical, functional and neurochemical maturation of pain pathways is well developed in fetus and neonates. Various physiological and behavioural responses to painful stimuli in neonates substantiate their ability to feel pain. Biological effects of pain are systematically studied in human fetus and neonates. Pain expressions in the newborn not only reflect tissue damage but are a function of ongoing behavioural state. The ultimate aim should be to keep neonates free from pain and other stressful stimuli as far as possible, by advocating minimal handling protocol, giving comforts after painful procedures, local anesthesia while carrying out painful procedures like cutdown and insertion of chest tubes, and if a baby is ventilated fentanyl and/or midazalam infusion must be carried out during initial periods of ventilation. PMID- 10798045 TI - Non-invasive monitoring of carbon-dioxide in newborns and children. PMID- 10798046 TI - Childhood tuberculosis--issues and challenges. PMID- 10798047 TI - Neonatal cerebral infarction: USS or CT for imaging? AB - Neonatal cerebral infarction or neonatal stroke is the second most common cause of neonatal seizures in term new born babies. It remains one of the least recognised entities in the neonatal text books and the awareness of its existence among general practitioners and practising paediatricians is very low. Cranial Ultrasound Scan (USS) is the standard imaging used by neonatologists and many neonatal intensive care units have their own scanners. However, USS may give a false sense of security especially in the term neonates and we report a case of neonatal cerebral infarction here and the need for computerised tomography (CT) for imaging. PMID- 10798048 TI - Plasmodium ovale malaria in Delhi. AB - A case of ovale malaria in a child from Delhi is reported. Urban malaria ecotypes caused by P. ovale has never been seen before. Characteristic morphological features of the parasite in stained blood film confirmed its identification. PMID- 10798049 TI - Stiff man's syndrome. AB - Myositis ossificans progressiva is an autosomal dominant disease resulting in progressive ossification and skeletal deformities, mainly in the connective tissue of muscle. The diagnosis is based on clinical and radiological findings and demonstration of skeletal malformations. We report a singular case of advanced myositis ossificans progressiva in a fourteen year old male and analyse the natural history, etiopathology, treatment alternatives and prognosis of this crippling disorder. PMID- 10798050 TI - Congenital tuberculosis. AB - A 29 day old male infant presented with a history of fever, cough, increasing respiratory distress and abdominal distension from the 10th day of life. Examination revealed failure to thrive, marked tachypnea, pallor, hepatosplenomegaly and harsh vesicular breath sounds. Chest skiagram showed extensive broncho-pneumonic changes. As the infant did not respond to antibiotics, he was investigated for tuberculosis. The gastric aspirate smear showed plenty of acid fast bacilli (AFB) and culture showed M. tuberculosis growth. Endometrial biopsy of the mother showed tuberculous granuloma and acid fast bacilli and culture of the aspirate from endometrium grew M. tuberculosis. The need for endometrial biopsy of mothers of infants with congenital tuberculosis is highlighted. PMID- 10798051 TI - Familial glucocorticoid deficiency, alacrimia and achalasia--Allgrove syndrome. AB - We report three brothers with Allgrove syndrome. All three had evidence of adrenal insufficiency and deficient tear production, though neither of them had achalasia, the third component of the disorder at the time of this report. Neurological abnormalities were present in the index case. The younger siblings were neurologically normal. The familial association of achalasia, alacrimia and adrenal insufficiency, rather than being fortuitous, is a distinct clinical entity. PMID- 10798052 TI - Single CT (ring) lesion in epilepsy patients: a new observation. PMID- 10798053 TI - An epidemic of acute haemorrhagic conjunctivitis in school children. PMID- 10798054 TI - Venous thromboembolism. AB - Incidence of venous thromboembolism (VTE) in children is rapidly rising. Frequent use of central venous lines (CVLs) in children with a variety of disease processes has contributed to increased incidence of VTE. In addition, increased detection of VTE has been possible due to the availability of sensitive imaging studies such as Color Duplex/Doppler ultrasound, lineogram and venogram. Heightened awareness of congenital prethrombotic disorders may help identify children at higher risk of thrombosis. Anticoagulation with heparin, low molecular weight heparin (LMWH) and oral anticoagulant such as warfarin are used for the treatment of VTE. Thrombolytic therapy may be indicated in some cases. Clinical presentation, management and pathogenesis including role of congenital prethrombotic disorders in pediatric VTE are discussed. PMID- 10798055 TI - High dose methylprednisolone therapy in nephrotic syndrome. AB - This study was done to determine the efficacy of oral high dose methylprednisolone (HDMP) therapy in the treatment of childhood nephrotic syndrome (NS). Fifteen patients were enrolled in the study. Patients were arbitrarily divided into two groups. Group I received prednisolone (daily 60 mg/m2 for 4 weeks, 45, 30, 20, 10, 5 mg/m2 on alternate days for 4 weeks) and group II received HDMP (30 mg/kg/d for 3 days, 20 mg/kg/d for 4 days, 10 mg/kg/ for a week, before 9 am, orally). The patients were followed-up for a duration of 38.0 +/- 5.5 months (range 24-68 months) in group I and 42.1 +/- 5.5 months (range 16-72 months) in group II. No significant difference was obtained in the duration of remission between both groups (p > 0.05), while HDMP induced early remission than prednisolone (p < 0.05). The mean relapse rate was 0.8/year in group I and 0.8/year in group II (p > 0.05). Although, the number of the patients were limited in the study it can be recommended that patients with NS can be treated with oral HDMP therapy as an alternative to standard oral prednisolone therapy. PMID- 10798056 TI - Sonographic assessment of renal volume in Indian children. AB - In an attempt to making norms of renal volume (RV) in Indian children, 240 children (137 boys and 103 girls) below 5 years of age were subjected to sonographic examination in supine position. Left RV in female children increased from 18.04 mm3 in early infancy to 48.36 mm3 in 4-5 year age group. Similar increase was observed in the right RV. There was no significant left vs. right difference in RV in boys and girls (p > 0.05). Similarly, the RV did not differ significantly in both sexes (p > 0.05, in all age groups except 4-5 years, p = 0.04). PMID- 10798057 TI - Using government schools to monitor iodine content of salt at household level in Delhi. AB - The Government had banned the sale of un-iodized salt in Delhi in 1989 as a step towards elimination of iodine deficiency disorders. The present study was done to detect the presence of iodine in salt samples every month from households of the students of government middle schools in Delhi using spot testing kits. One section each, from the classes six to eight of the thirty selected middle schools was chosen randomly. The children were asked to bring salt from their homes on a prefixed day. The salt samples were examined for the presence of iodine by a spot testing kit. This was repeated every month during the study (August '94 to February '95) among the same students in each school. A total of 16,596 salt samples were collected. Of this 12,736 (76.7%) tested positive for iodine and the rest (23.3%) of the salt samples did not contain iodine. The proportion of children who brought the salt samples varied between 53% to 72% in different months. As the study progressed, the trend showed more samples being tested positive for iodine. The differences in districts were not significant. Un iodized salt thus still continues to be available in the households of the government school children of the National Capital Territory of Delhi. The use of rapid testing kits in schools is a practical and feasible way to enhance the monitoring activities of the government. PMID- 10798058 TI - Assessment of status of salt iodization in Delhi. AB - The National Capital Territory (NCT) of Delhi was identified as an iodine deficiency endemic area in 1980. The government of NCT of Delhi banned the sale of non-iodised salt since 1989. The present study was aimed to estimate the iodine content of salt consumed in the households of the state. Thirty clusters were selected using population proportionate to size cluster sampling procedure. In each identified cluster one primary school was randomly selected. In each school, 60 salt samples were collected from an equal number of school children. The iodine content of a total of 1854 salt samples collected was analyzed using the standard iodometric titration method. Forty one per cent of families consumed salt with an iodine content of less than 15 ppm. Salt with nil iodine content was consumed only by 1.4% of the beneficiaries which indicated successful implementation of universal salt iodization programme in the state. PMID- 10798059 TI - Awareness of brain death and organ transplantation among high school children. AB - The objective of this study was to assess the awareness of the concepts of brain death and organ transplantation among high school children. One hundred and eighty eight students of class 12th of a reputed public school were studied. Structured questionnaires were used to assess their knowledge in various aspects of brain death and organ transplantation. Following the questionnaire, they were provided with educational information on brain death and organ transplantation. This was followed by similar questionnaires to assess any change in awareness of brain death and organ transplantation. Results spoke of widespread awareness and acceptance of organ transplantation in the high school children. However, the awareness of various aspects of brain death was quite low. There was significant increase in awareness and acceptance of brain death after educating the students. Education about various aspects of brain death, its immense importance for organ donation and legality of brain death needs to be highlighted. PMID- 10798060 TI - Endocrinological parameters in experimental unilateral undescended testis. AB - It is a common clinical practice to estimate FSH, LH and testosterone levels in patients with unilateral undescended testis (U/L UDT) as a prognostic pointer to fertility potential. Is this practice correct? To ascertain this aspect, new born rats were operated to create experimental U/L UDT by gubernaculectomy and anchoring the gubernaculum to anterior abdominal wall. Fertility and hormone levels were evaluated later in adult life. Though fertility of rats with U/L UDT was significantly less (p < 0.01) than the controls, no significant alterations were found in the levels of serum testosterone, FSH and LH. Even though variations in the hormone levels may be responsible, to a certain extent, for the decrease in fertility potential in U/L UDT, estimation of sex hormone levels in U/L UDT is not a sensitive indicator of fertility potential. U/L UDT may additionally be affecting fertility through non-endocrinological mechanisms. PMID- 10798061 TI - Pediatric nephrology. PMID- 10798062 TI - Acute glomerulonephritis. AB - Acute glomerulonephritis (AGN) manifests with abrupt onset of hematuria, facial edema, hypertension and impairment of renal function. The commonest form of AGN in developing countries is that following a beta hemolytic streptococcal infection where the glomerular injury is mediated by deposition of immune complexes. In the usual patient with moderately severe poststreptococcal AGN (PSAGN) the above-mentioned features are present However, gross or microscopic hematuria may be the only abnormality. A similar picture may occasionally be produced by a variety of infections (when GN is referred to as post-infectious and the mechanism of glomerular damage and the renal histology are similar to that in PSAGN), primary renal glomerular disorders (e.g. membranoproliferative GN, IgA nephropathy), collagen vascular diseases (systemic lupus erythematosus), systemic vasculitis (Henoch Schonlein purpura) and hereditary nephritis and some nonglomerular conditions. PSAGN may also present with one or more of its complications such as profound volume expansion with heart failure and hypertensive encephalopathy. PSAGN resolves rapidly and has an excellent prognosis. Patients with severe renal involvement and life threatening complications need expert supportive management. AGN with associated systemic features or very pronounced azotemia, nonstreptococcal AGN and unresolving GN need prompt, appropriate evaluation that often includes a renal biopsy. If extensive crescentic changes are found (crescentic GN), aggressive immunosuppression will be necessary. PMID- 10798063 TI - Asymptomatic hematuria in childhood: a practical approach to evaluation. AB - The differential diagnosis of hematuria with or without proteinuria is extensive, and isolated hematuria is a common problem in children and adolescents. Extensive evaluation is often necessary for the child presenting with macroscopic plus microscopic hematuria including nonglomerular and glomerular etiologies, while children with only isolated microscopic hematuria can generally be followed after baseline evaluation to rule out infection, hypercalciuria, familial hematuria, sickle cell disease, post-streptococcal glomerulonephritis (GN), and structural abnormalities (cysts, stones, obstruction, Wilms tumor). Children with the combination of hematuria and proteinuria require rapid systematic evaluation, generally including renal biopsy, except in cases where post-streptococcal GN can be clearly documented. Post-streptococcal GN occurs 7-21 days after a streptococcal infection, is associated with an acute fall in C3 levels with return to normal by approximately 8 weeks, rarely causes acute renal failure, and in children has a pattern of gradual resolution of hypertension, hematuria, and proteinuria over a course of 6-12 months. PMID- 10798064 TI - Lupus nephritis in children. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease with varied clinical manifestations. Children and adolescents comprise one-fourth of affected patients with SLE and 40-80% of them have renal involvement. Lupus nephritis (LN) may present with mild urinary abnormalities or fulminant acute nephritis and renal failure. Diffuse proliferative glomerulonephritis (WHO class IV) is the predominant histological presentation in children and more common in boys than girls. This probably is one of the main reasons for the high mortality reported in the initial studies. Early diagnosis and aggressive treatment have led to improvement prognosis in these children. Cytotoxic therapy including intravenous cyclophosphamide has a definite role in the management WHO class IV and occasionally class III lupus nephritis. Prolonged steroid and cytotoxic therapy may lead to significant toxicity. PMID- 10798065 TI - Management of acute renal failure. AB - Acute renal failure (ARF) is defined as an abrupt decline in the renal regulation of water, electrolytes, and acid-base balance. It continues to be an important factor contributing to the morbidity and mortality of critically ill infants and children. The frequency of specific diseases that result in ARF differs among different age-groups and geographical areas. The common causes in Indian children include hemolytic uremic syndrome, acute tubular necrosis, glomerulonephritis and urinary tract obstruction. Though the hallmark of renal failure is oliguria, there is increasing recognition of non-oliguric ARF often associated with the use of nephrotoxic drugs. The basic principles of management are avoidance of life threatening complications, maintenance of fluid and electrolyte balance and nutritional support. Specific management of the underlying disorder is possible only in a minority of cases. All the major dialysis modalities--peritoneal dialysis (PD), hemodialysis (HD) and continuous hemofiltration--can be used to provide equivalent solute clearance and ultrafiltration. Peritoneal dialysis requires minimal equipment and infrastructure, and is easy to perform; this makes it the favoured modality in developing countries where resources for HD or continuous therapies may not be accessible. However, continuous hemofiltration is an excellent alternative to PD in patients with ARF and severe fluid overload. The prognosis of children with renal failure depends on the underlying condition and associated medical complications. PMID- 10798066 TI - Evaluation and treatment of chronic renal failure. AB - Chronic renal failure (CRF) is the irreversible deterioration of renal function that gradually progresses to end stage renal disease (ESRD). The chief causes of CRF include obstructive uropathy, primary glomerular diseases, reflux nephropathy and hypoplastic or dysplastic kidneys. Progressive hyperperfusion and hyperfiltration causes increasing glomerular injury and further renal damage. Symptoms of CRF are usually seen when GFR is between 10-25% of normal. Children with severe CRF often suffer from failure to thrive, growth retardation, acidosis, anemia and renal osteodystrophy. Management of CRF aims at retarding progression of renal damage and treatment of complications related to renal dysfunction. Measures suggested to retard progression include protein restriction, strict control of hypertension, use of angiotensin converting enzyme inhibitors and control of hyperlipidemia. Appropriate amounts of protein and calories are recommended to prevent growth failure. Nutritional supplements are often required. The availability of recombinant erythropoietin, calcitriol and human growth hormone has significantly improved the management of these patients. Once ESRD supervenes, renal replacement therapy in the form of chronic peritoneal or hemodialysis and transplantation is necessary. PMID- 10798068 TI - Renal transplantation in infants and children. AB - Renal transplantation is the treatment of choice in children with end stage renal disease. Advances in organ retrieval and preservation, improved surgical techniques and postsurgical care, newer immunosuppressive drugs and prevention and treatment of infections have significantly improved survival of the renal allograft. The absolute requirements for a transplant are compatible blood group and a negative cytotoxic crossmatch. HLA identical grafts have a longer half-life than those that are less well matched. The immunosuppressive drugs most often used are cyclosporin A (or tacrolimus), azathioprine (or mycophenolate mofetil) and prednisone. Complications following transplantation include episodes of acute rejection, serious bacterial and viral infections, hypertension and recurrence of primary disease in the allograft. Each centre must have standard protocols for pre-transplant evaluation, and monitoring during surgery and in the post operative period. Socio-economic factors should be evaluated before offering renal transplantation to children in developing countries. PMID- 10798067 TI - Dialysis therapy in end-stage renal disease. AB - The prognosis for children on dialysis has improved significantly in the past two decades. Much of this improvement can be attributed to the realization that adequate nutrition is a critical element of dialysis therapy and long-term morbidity and mortality in the dialysis population are closely linked to the nutritional state. Recommendations for nutritional intake have been formulated for infants and children with end-stage renal disease that take into account not only the metabolic derangement but also the effect of the dialysis treatment itself on the gain and loss of nutrients. In addition, the relationship between nutritional intake and the "dose" of dialysis is becoming clearer. Increasing experience in pediatric dialysis is enabling better selection of the mode of dialysis for children of different ages. The realization that the permeability of the peritoneal membrane is different from individual to individual has led to customized dialysis prescriptions with a consequent increase in the efficacy of peritoneal dialysis. When combined with improvements in therapy of medical complications of chronic renal failure, including the availability of synthetic erythropoetin++ and growth hormone and the management of renal osteodystrophy, dialysis is becoming a fully-functional tool in the management of children with end-stage renal disease. PMID- 10798069 TI - Neuropsychiatric aspects of pediatric thyrotoxicosis. AB - The neurobehavioural and neuropsychiatric changes associated with thyrotoxicosis are multiple and varied. This association is well recognised although the true incidence of neuropsychiatric symptoms in thyrotoxicosis is not known. Review of available literature suggests that frank psychiatric symptoms in thyrotoxicosis may be in the order of 10%. In pediatric thyrotoxicosis the neuropsychiatric symptoms may be pronounced and may antedate the medical diagnosis by six months to one year. One of the classic presentation is deterioration in school performance. Frequently noticed cognitive and behavioural abnormalities in pediatric thyrotoxicosis are hyperactivity, irritability or anxious dysphoria, and problems of attention. Successful treatment of thyrotoxicosis usually leads to resolution of the major mental disturbances associated with it and delayed treatment possibly results in enduring neuropsychiatric problems. Awareness of neuropsychiatric symptomatology can help in early detection and appropriate management of children with thyrotoxicosis. PMID- 10798070 TI - Long term survival in a young girl with renal cell carcinoma. AB - A young girl with an uncommon renal tumour is the subject of this communication. A diagnosis of renal cell carcinoma was established post-operatively. Combined modality treatment including chemotherapy and radiotherapy has helped achieve a disease free survival of nearly 5 years. PMID- 10798071 TI - Foreign body airway in neonates. AB - Two cases of foreign body in neonates less than one month of age are reported. Although foreign bodies in neonates are unknown but the possibility should not be overlooked even in neonates especially with sudden onset of respiratory distress, cough or hoarseness in absence of fever. PMID- 10798072 TI - Fetal hydantoin syndrome with rheumatic valvular heart disease. AB - Research has shown that anticonvulsants are teratogens and pose a risk for fetal malformations. Though Fetal Hydantoin Syndrome (FHS) was first reported by Langhman and others, wide phenotypic variability of this syndrome has lead many clinicians to question its very existence. We report a twelve year old girl with FHS with rheumatic valvular heart disease. PMID- 10798073 TI - Retro-peritoneal cystic lymphangioma in association with fetal hydantoin syndrome. AB - Antiepileptic drugs are known to be teratogenic. Use of phenytoin during pregnancy can cause various congenital malformations leading to 'fetal hydantoin syndrome'. One such case reported is unique in the sense that it occurred with retroperitoneal cystic lymphangioma, itself a rare condition. Such an association is not described elsewhere. PMID- 10798074 TI - Anterior tibial artery pseudoaneurysm. AB - Traumatic pseudoaneurysms are rare in civilian trauma, more so in childhood. A case of anterior tibial artery pseudoaneurysm is described which was successfully treated non surgically by percutaneous coil embolisation, thus questioning the need for surgery in peripheral pseudoaneurysms. PMID- 10798075 TI - Partial 1q and 21p trisomies in a male child due to maternal t(1;21). AB - Proband 7 years old male child referred for cytogenetic investigation revealed 47, XY + der (21), t(1;21) (q32;q11) mat. PMID- 10798076 TI - Haematuria in haemorrhagic disease of newborn. PMID- 10798077 TI - Pediatric transfusion therapy: practical considerations. AB - Over the past decade, safety of blood has increased tremendously because of better donor screening as well as testing of the units for transmissible diseases. Component therapy has allowed more effective and economic use of blood. Whole blood is rarely used; instead, packed red cells, platelets, and fresh frozen plasma (FFP) are the most common components used. These products are further refined using irradiation and microaggregate filters and in the case of FFP, viral inactivation. Irradiation prevents transfusion-associated graft versus host disease, whereas microaggregate filters remove leukocytes, decreasing the rates of alloimmunization, febrile nonhemolytic (FNH) reactions, and cytomegalovirus (CMV) transmission. Autologous donation in older children probably provides the safest blood as far as transmissible diseases are concerned. More families request a directed donation and solicit physician help in deciding as well as making arrangements for autologous and/or directed donations. Transfusions of blood and blood components in children are often challenging and require a knowledge of physiologic changes in hemoglobin and blood volumes during different ages. The unique needs of neonates, immunocompromised patients, and patients with congenital hemolytic anemia (sickle cell, thalassemia) mandate that the pediatrician have an appropriate knowledge of transfusion volumes and choice of blood product as well as indications for transfusion. PMID- 10798078 TI - Assessment of sensorial oral stimulation in infants with suck feeding disabilities. AB - A non-randomized single blind study was undertaken to determine the clinical and physiological changes in suck feeding after sensorial oral stimulation, in fourteen patients age 9 to 210 days old with sucking alterations. Patients lacked at least one of the five oral reflexes, plus two or more abnormal sucking sings or at least one abnormal sucking sign, plus two or more abnormal oral reflexes. Oral sensorial therapy was performed thrice daily for five days. The number of absent oral reflexes, number of abnormal sucking signs, volume of milk for nursing and sucking rate, were registered. Differences of medians were tested using Freidman's test and differential of proportions using Cochran's Q test. After therapy, oral reflexes were recovered (2, 0-4 vs. 5,5-5, p = 0.0000, median rank of absence oral reflexes) and the number of abnormal sucking signs decreased (6,1-9 vs. 1, 0-4; p = 0.0000). There were statistically significant improvements in patients who had lost launch up nipple ability (p = 0.005), delay at the beginning of sucking (p = 0.0022), drawing of milk from the mouth (p = 0.0001), cyanosis (p = 0.0084), weaning (p = 0.0004) and prolonged sucking (p = 0.0038). Even in patients with moderate improvement, no statistical differences were observed in nipple rooting (p = 0.09) and coughing (p = 0.09). No changes were observed in patients who had cried (p = 0.31) and spitted (p = 0.51) during feeding. At the end of therapy, volumes of consumed milk were increased at each feeding (10 ml, 0-40 vs. 50 ml, 25-60; p = 0.0001). Sucking rates also increased (22 sucks/minute, 10-35 vs. 40.5, 35-48; p = 0.0044). Oral sensorial and motor stimulation normalise oral motor reflexes, diminish the clinical abnormal sucking signs and increase milk volumes ingested for nursing. PMID- 10798079 TI - Development and assessment of a screening test for detecting childhood disabilities. AB - The paper outlines the development and assessment of a screening test for broad based identification of major disabilities in children under 6 years of age. The Disability Screening Schedule (DSS) has been developed which should act as a one time screen for all major disabilities viz. physical, motor, sensory and mental retardation. The DSS was developed after reviewing a number of existing screening instruments. It was pilot tested in 3 phases and suitably modified. Nineteen AWW received a short training and used the DSS to screen children with disabilities in their respective areas. It was field tested by administering it on 3560 children (0-6 years) drawn from nine urban slums of South Delhi. The workers used the DSS and identified 245 children as having an impairment/at risk conditions and 3315 children were reported as normal. The investigator cross checked 219 'impaired' and 536 'normal' children. On the basis of the review exercise, the DSS was validated and was found to have a sensitivity of 0.89 and a specificity of 0.98. The DSS is a short questionnaire, and the administration time is about 5 minutes. PMID- 10798081 TI - IgG subclasses in wheezing infants. AB - The wheezing infant is a common but difficult patient to approach diagnostically. The prevalence of IgG subclass antibody deficiency in wheezing infants is still controversial. We studied serum concentration of IgG subclasses in 38 wheezing infants (aged 6-24 months who had not received systemic steroids before investigation) and in 30 healthy age matched control (aged 6-24 months). The prevalence of one or more IgG subclass deficiency was 31.6% in wheezing infants and 26.7% in controls. There was no significant difference in prevalence of IgG subclass deficiency between patients and controls (p > 0.05). The mean concentration of IgG subclasses in patients were compared with controls. There was no significant difference in mean serum concentration of IgG1, G2 and G3 subclasses. But there was a trend towards higher concentrations of IgG4 in wheezing infants and this difference for IgG4 was significant (p < 0.01). However, IgG subclass deficiency was found in 25% and 36.4% of wheezing infants who had experienced from two to four and five or more wheezing episodes in two years, respectively (p > 0.05). These findings suggest that wheezing in infancy is not associated with IgG subclass deficiency and in wheezing infants low IgG subclass levels do not increase the frequency of wheezing. PMID- 10798080 TI - Immunoglobulin isotypes in childhood asthma. AB - Immunoglobulin isotypes (IgG, IgA, IgM, IgD, IgE) in serum were investigated in 64 Libyan children with mild to moderately severe asthma (age: 1-12 years; sex: 39 males, 25 females) (Group A) and in 57 healthy Libyan children (age: 1-12 years; sex: 30 males, 27 females (Group B). The patients were classified according to age into three groups (A1: 1-3 years; A2: > 3-5 years; A3: > 5-12 years); according to disease activity into two groups (AA: active disease; NA: inactive disease); and according to age plus disease activity into six groups (AA1, NA1; AA2, NA2; AA3, NA3). The healthy children were also divided according to age into three groups (B1: 1-3 years; B2: > 3-5 years; B3: > 5-12 years). IgG, IgA, IgM and IgD were measured by radial immunodiffusion method and IgE was estimated by enzyme immunoassay technique utilizing immunokits from bioMerieux, France. Serum levels of IgG, IgD and IgE were elevated significantly in patients compared to controls (A vs B: p < 0.05) while IgA and IgM levels were normal (p > 0.05). IgG and IgD levels were raised in A3 (p < 0.05), while IgD levels were raised in both A2 and A3 (p < 0.05) and IgE was elevated in all age groups (p < 0.05). However, IgG was elevated significantly in AA only, while IgD and IgE levels were high in both AA and NA (p < 0.05) and IgE was even considerably higher in AA compared to NA (p < 0.02). Further elevated levels were observed for IgG in AA3 only (p < 0.05), for IgD in NA2 (p < 0.01), AA3 (p < 0.01) and NA3 (p < 0.05) and IgE was much higher in patients with active disease than with inactive disease in all age groups (p < 0.05). The fact that asthmatic attack in majority of our patients can be explained as mediated through IgE and the possibilities that IgG and IgD may play roles as aetiopathogenetic or protective regulatory factors in childhood asthma are discussed. PMID- 10798082 TI - Results of the gastroesophageal reflux assessment in wheezy children. AB - Gastroesophageal reflux (GER) is implicated in the pathogenesis of respiratory symptoms in childhood. It should be taken into account especially in the differential diagnosis of children presenting with wheezing. Although, oesophageal pH monitorization has been reported to be the best technique in the evaluation of GER, radionuclide studies have also been shown to be very sensitive recently. In this study, 82 children presenting with recurrent wheezing (n = 74) and/or vomiting (n = 28) (mean age 17.4 months; range 3-48 months) were evaluated. GER scintigraphy was performed to determine the frequency of GER. GER was determined in 18 of the 82 cases (21.9%). The GER was found in 21.1% of children with recurrent wheezing and in 16.6% of children suffering from recurrent vomiting. GER scintigraphy should be kept in mind in the evaluation of children with the complaint of recurrent wheezing since it is a noninvasive and easily applicable method. PMID- 10798083 TI - Role of inhaled nitric oxide as a selective pulmonary vasodilator in pediatric cardiac surgical practice. AB - Our aim was to assess the role of inhaled nitric oxide (NO) therapy in post operative cases of congenital heart defects who developed pulmonary arterial hypertensive (PAH) crisis and had no response with conventional management. From February '95 to January '97, inhaled NO therapy was used in 21 children. Age ranged from 2 months to 9 years (mean 5.6 years) and duration of therapy ranged from 1 to 13 days. Of 21 patients, 17 responded well with 5-20 ppm while 4 did not. The preoperative mean pulmonary systolic pressure was 88 mm Hg against mean systemic pressure of 96 mm Hg. Post operatively, their PA pressure reduced to 62 mm Hg, with systemic pressure of 98 mm Hg. After using inhaled NO, PA pressure dropped to 24 mm Hg (mean systolic) (p < 0.007), after excluding the non responders. Of 4 non responders, two died due to irreversible pulmonary vascular disease and remaining two died due to residual defects. The study shows that inhaled NO is a selective pulmonary vasodilator, which is useful in postoperative PAH crisis and also reduces the transpulmonary gradient in single ventricle repair cases. It is safe and effective for prolonged use. It is very useful in Indian perspective, when more number of cases with congenital heart defects (CHD) along with severe PAH are encountered routinely. PMID- 10798084 TI - Permanent neonatal diabetes mellitus: epidemiology, mode of presentation, pathogenesis and growth. AB - Permanent neonatal diabetes mellitus (PNIDDM) is a rare form of IDDM with unclear etiology and pathogenesis. We determined the incidence and prevalence rates and studied the clinical and biochemical features of PNIDDM in the Sultanate of Oman. The mean incidence rate during the study period from January 1989 to December 1994 was 1.788 +/- 0.82 per 100,000 live births per year. At the end of December 1994 the prevalence rate was 2.4 per 100,000 children below the age of 5 years. They constituted 41.6% of all cases of IDDM in this age group. Diarrhoea, fever, lethargy, poor feeding and failure to thrive were the most common presenting symptoms. Dehydration and tachypnoea were the most common signs. All patients who developed IDDM during the neonatal period had intrauterine growth retardation and 4.5 presented with diabetic ketoacidosis (plasma glucose 37 +/- 9 mmol/L, pH 7.12 +/- 0.1). Hypertriglyceridemia was a constant feature (19.4 +/- 4.8 mmol/L). They were products of consanguineous marriage with significantly high prevalence of IDDM and NIDDM in their family members. None of the infants had clinical or immunological evidence of congenital viral infection. Three of the five children had HLA-DR2, the diabetes resistance alleles. C-peptide secretion was absent during and after metabolic control of hyperglycemia in all the studied infants and none had circulating islet cell antibody at presentation or during the first year after diagnosis. Despite marked growth retardation at birth, there was a significant improvement of growth after initiating insulin therapy. Four of the 5 patients had normal developmental milestones, one had mild developmental delay following a severe and prolonged attack of hypoglycemia. None of the patients had exocrine pancreatic deficiency. In summary, the very high rate of parental consanguinity, occurrence in both sexes and in two siblings in the same family, absence of islet cell antibodies and the presence of HLA-DR2 loci in 3/5 of patients suggest that PNIDDM is a different disease process to standard IDDM in childhood and an autosomal recessive mode of transmission. PMID- 10798085 TI - Evaluation of bone mineral density in children with diabetes mellitus. AB - Multiple studies have documented reduction in peripheral bone mass in children with insulin dependent diabetes mellitus (IDDM). In this study, the bone mineral density (BMD) of the lumbar vertebrae (L2-L4) was measured by dual photon absorptiometry in 14 female and 16 male diabetic patients of age 11 to 16 years with varying clinical duration. Twenty three children between 11 to 16 years with normal anthropometric measurements between 10th and 97th percentile and no known history of metabolic bone disease served as a control group. BMD values, weight, height, body mass index, metabolic, biochemical and growth parameters of the study group were compared with those of the control group. BMD (L2 AP 0.732 +/- 0.15 gm/cm2, L2 lateral 0.534 +/- 0.09 gm/cm2 in the study group and 0.812 +/- 0.63 gm/cm2 and 0.619 +/- 0.20 gm/cm2 in the control group) and osteocalcin (10.10 +/- 3.40 ng/ml and 23.12 +/- 2.74 ng/ml in diabetes and control respectively) levels were significantly lower in diabetic patients (p < 0.05, p < 0.01 respectively). Within the study group BMD correlated positively with age but not with the duration of the disease nor with the level of metabolic control. PMID- 10798086 TI - Serum lipid and lipoprotein composition in infants of diabetic mothers. AB - Diabetes mellitus (DM) alters carbohydrate and lipid metabolism to a great extent. This study was planned to determine whether infants of insulin dependent and gestational diabetic mothers have abnormal lipid metabolism. Three groups of newborns were included in the study; group I consisted of 7 infants of diabetic mothers (IDM) with insulin dependent DM (Type 1 DM), group II of 18 infants of gestational diabetic mothers and group III of 20 control neonates whose mothers had no history of DM. Total cholesterol (TC), triglyceride (TG) and high density lipoprotein-cholesterol (HDL-C) values in groups I and II were no different compared to those in group III (p > 0.05). However, low density lipoprotein cholesterol (LDL-C) and LDL-C/HDL-C ratio were similar between groups I and II (p > 0.05) but significantly higher in both infants of type 1 diabetic mothers and gestational diabetic mothers compared to control infants (p < 0.05). Apolipoprotein A-I (Apo A-1) and apolipoprotein B (Apo B) levels, Apo A-I/Apo B and HDL-C/Apo A-I ratios were similar in between groups. However, Apo B/LDL-C ratio was significantly lower in groups I and II compared to control group (p < 0.05). In conclusion, diabetes in pregnant women causes a tendency to LDL hypercholesterolemia in the offspring. These infants should be longitudinally followed up to assess whether this observation imposes an increased risk for atherosclerosis for advanced ages. PMID- 10798087 TI - Behavioural assessment of children and adolescents. AB - This article is an overview for pediatricians who conduct behavioural assessments of children and adolescents. It identifies the most common behavioural problems encountered by pediatricians and brief descriptions of selected tests that are administered by psychologists and other trained mental health professionals. Also covered are suggestions for the pediatrician on conducting diagnostic interviews and information regarding referral decision making. PMID- 10798089 TI - Reproductive health: an international perspective. AB - The twentieth century began with approximately 1.6 billion human beings in the world and, with an increase of 96 million people per year, will end with over 6 billion people. Unless this trend is directly confronted by the world governments and their citizens, there will be nearly 8 billion by 2025 and over 11 billion by 2050. Thus, reproductive health is and will remain an issue of critical importance for all countries to realize and study. Part of this trend is because of the many pregnancies which occur in adolescents around the world. A number of issues have developed this century contributing to the many pregnant teenagers. For example, India has one of the world's largest populations of teenagers--over 23 million, representing over 26% of the total in the world. Effective methods for contraception and sexually transmitted diseases prevention are available, but not to all sexually active humans. However, many barriers to effective contraception exist around the globe. This article discusses some of them and also reviews use of contraceptive methods in various countries. These include oral contraceptives, emergency contraceptives, injectable and implantable contraceptives, intrauterine devices, barrier contraceptives and others. Reproductive health remains a critical, universal issue for all humans in the world. We all must examine the many pitfalls to controlling the world's populations, including lack of sex education, limited access to effective contraceptives, and others. We cannot afford to let the population continue unchecked. Effective strategies are needed at this time; otherwise, the population will continue to run out of control, negatively damaging the world for the coming generations. There is need to leave a positive, and not negative legacy for the next generation. PMID- 10798088 TI - Disorders of cognition, attention and learning. AB - The inability to successfully navigate the educational system can cause serious problems for children, their parents, the Indian society, and the world at large. When children are required to engage in academic exercises which require attention, specific cognitive abilities and processes, and demonstrations of learning, a subset of these children are unable to perform due to some type of intellectual, emotional, behavioural, physical, or environmental deficit. Any combination of these factors can result in school failure. Although, resources for remedial or special education services in Southeast Asia and India are particularly meager, efforts must be directed towards the retention and achievement of those children who do enroll in primary and secondary schools. Learning disability represents a major form of impairment in society, and much more needs to be done to bring about a general awareness of its consequences. An urgent priority is the education of physicians and teachers about disorders of cognition, learning, and attention. PMID- 10798090 TI - Enuresis: current concepts. AB - Incontinence disorders are an important group of problems that clinicians manage in children and adolescents. This paper reviews the physiology of micturition, the epidemiology of enuresis, etiologic concepts of incontinence disorders, a general clinical approach to enuretic patients, laboratory evaluation and general principles of management. Neurological and nephrological concepts of enuresis are emphasized in this discussion. PMID- 10798092 TI - Psychosocial aspects of chronic illness in adolescence. AB - Success in the survival of children with significant medical conditions has resulted in a dramatic increase in the prevalence of adolescents with chronic illness. Unfortunately, the traditional biomedical approach does not prepare practitioners for the complexities of managing chronic illness, because numerous psychosocial factors are involved. This article will address the need for integrating psychosocial issues into the assessment and treatment of the chronically ill adolescent. Assessment of the teen should incorporate an understanding of the developmental stages of adolescence, a review of how these developmental tasks are being accomplished, a discernment of individual characteristics, and knowledge of the family and community to which they belong. Management should take into account the phases of illness (acute, chronic or terminal) and encourage empowerment of the adolescent and family in decision making. A team approach that is community-based, comprehensive, and culturally appropriate is ideal. PMID- 10798091 TI - Encopresis. AB - Encopresis is fecal soiling associated with functional constipation in a child. Constipation and encopresis are common problems in children. Encopresis is most common between ages 3 and 7 years. Infants and pre-school children present with a history of constipation and withholding maneuvers. The school-age child may have constipation and fecal soiling for some time prior to detection. In some children encopresis is associated with enuresis and urinary tract infection. Family education is the essential first step in management, followed by disimpaction of stool and complete evacuation of the rectum. Reaccumulation of stool should be prevented by appropriate use of laxatives and stool softeners. This is followed by a gradual weaning of the laxative regimen and instituting toilet training. Relapses may occur. Up to 50-60% of children achieve acceptable bowel control, free of soiling, within a year. PMID- 10798093 TI - Progressive pseudorheumatoid arthropathy of childhood. AB - Progressive pseudorheumatoid arthropathy of childhood (PPAC) described by Spranger et al is a rare autosomal recessive disorder. An 11 year-old girl was diagnosed as having PPAC at Ege University, Faculty of Medicine, Department of Paediatrics. Her complaints of painful joints, difficulty in walking and joint contractures began at the age of 3 years and she was treated for juvenile rheumatoid arthritis for 8 years. Her symptoms did not respond to nonsteroid anti inflammatory treatment. During her last hospitalisation period, she was reinvestigated. Radiological examination showed spondyloepiphyseal dysplasia, severe acetabular irregularity and osteoporosis. All the laboratory test results for rheumatoid arthritis were negative. The clinical and radiological findings of the patient are illustrated. PMID- 10798094 TI - Tar syndrome with unilateral absent radius and associated esophageal atresia: a variant? AB - We report a male child with Thrombocytopenia and Absent Radii (TAR) syndrome but with single absent radius and associated tracheoesophageal fistula which has never been reported till date. PMID- 10798095 TI - Unusual complication--VP shunt coming out per rectum and brain abscess. AB - VP shunt is the most common pediatric neurosurgical procedure. It has been shown to result in variety of complications, which may have devastating consequences. Discussed below is one of the rare complications and the strategy to manage it successfully. PMID- 10798096 TI - Foreign body in vagina--an uncommon cause of vaginitis in children. AB - An intravaginal foreign body of long duration can pose diagnostic dilemma in children. We present a case of eight and a half years old girl who was suffering from blood stained vaginal discharge for 3 years for which she was treated by few gynaecologists. A vaginal examination performed under general anaesthesia revealed a foreign body (lead pencil). In cases of pediatric vaginitis one should always look for foreign body in vagina. PMID- 10798097 TI - "Plasmodium ovale infection in Delhi" published in Indian Journal of Pediatrics Jan-Feb 1999; vol. 66 (1): 143-145. PMID- 10798098 TI - Isolated ocular relapse in acute lymphoblastic leukemia. PMID- 10798099 TI - Hypoxic-ischemic brain damage in perinatal age group. AB - Cerebral hypoxia-ischemia in the perinatal period continues to be a major contributor to chronic neurologic impairment in children worldwide. Extensive research conducted in the past several years has led to a better understanding of the mechanisms involved in hypoxic-ischemic brain injury. Based on this understanding, the major potential therapeutic approaches being studied include antagonists of excitatory amino acids, calcium channel antagonists, free-radical scavengers, nitric oxide synthase inhibitors, anti-inflammatory agents, trophic factors, and hypothermia. Several agents are in clinical trial phases in adults. However, safety concerns and close relationship between pathomechanisms of hypoxic-ischemic cerebral injury and normal developmental processes have contributed to the slow pace in the neonatal trials. Large multicenter trials including an adequate number of infants will be needed to evaluate efficacy of therapeutic interventions in this particular age group. A large number of risk factors that predispose to hypoxic ischemic injury have been identified. It is important to control these factors and prevent brain damage in the first place. This is especially true for developing countries where resources for treatment with newer agents (when they become available) are likely to be limited. Recent information regarding mechanisms of injury and potential therapeutic measures related to perinatal age are presented in this paper. PMID- 10798100 TI - Suitability of CANSCORE for the assessment of the nutritional status of newborns. AB - The suitability of Clinical Assessment of Nutritional Status Score (CANSCORE) for the assessment of foetal malnutrition among 372 local Hyderabad newborns was studied. Details of length, weight and body mass index (BMI) at birth were related to total CANSCORE which consisted of scores ranging from 1 to 4 based on the grades of clinical status of hair, cheeks, buttocks, chest, legs, back, neck, arms and skin of anterior abdominal wall. The correlation coefficients of CANSCORE with the length, weight and BMI of newborns indicated that score of hair was least correlated with nutritional status. Normal newborns were found to have the lowest prevalence of foetal malnutrition. In those with retarded measurements of length and weight or BMI, the prevalence of foetal malnutrition was higher. The newborns with retardation of both length and BMI had higher prevalence of foetal malnutrition. The feasibility of the suggested limits of CANSCORE for the foetal malnutrition was assessed. Values of 24 for total CANSCORE and of 22 for "Modified CANSCORE" (score excluding hair as a parameter) were found appropriate for the assessment of foetal malnutrition. Modified CANSCORE is a simple, rapid and quantifiable method for the assessment of foetal malnutrition in term newborns. PMID- 10798101 TI - Perinatal mortality at a tertiary care hospital in Punjab. AB - The present report is a comparative analysis of perinatal mortality rate (PNMR) over two different periods of seven years each viz. 1982-1988 and 1989-1995. Data of all the perinatal deaths in babies born at Christian Medical College and Hospital, Ludhiana from January 1989 to December 1995 was collected. The cause of death was ascertained by a detailed history, clinical examination and whenever possible, by autopsy and analysed by modified Wigglesworth's classification. The PNMR during both the study periods was exactly the same i.e. 74/1000. There was a significant decline in the early neonatal mortality rate from 32/1000 to 25/1000. This was mainly due to improved survival of preterms as there were better life support measures available in the latter part of study period. In contrast, the still birth rate increased significantly from 42/1000 to 49/1000, thus neutralizing the fall of neonatal mortality. There was no change in the pattern of causes of death. Macerated still births occurring mainly in growth retarded babies and asphyxia remained the major causes of death. Mere provision of health services is not going to decrease PNMR. There is a need to educate 'the ultimate' consumers i.e. the women, for better utilization of these services. There is also an urgent need to sensitize and involve the medical practitioners imparting obstetrical services for solving these issues. PMID- 10798102 TI - Attitude and practices regarding diarrhoea in rural community in Chandigarh. AB - The present study on diarrhoea, its prevalence, practices and awareness of mothers was conducted in 120 randomly selected households in the rural area of Chandigarh during monsoons in 1996. Inspite of access to safe drinking water and latrines in 83% and 74% of the households in the village respectively, the prevalence rate of diarrhoea in 181 under five children was observed to be 23.2%. Majority (88.1%) of children had treatment for diarrhoea whereas only half (54.8%) of children were given oral rehydration solution. 86.7% of the mothers were aware of ORS but only 18.7% could tell the correct method of its preparation. A large number of respondents implicated a variety of food items responsible for diarrhoea and restricted them during the episodes. PMID- 10798103 TI - Measles immunisation coverage in urban slums. AB - A cross-sectional, community-based study was conducted in 32 slums of Udaipur city to assess the prevalence of missed opportunities for measles immunization, reason for their occurrence and potential impact of avoiding them on measles immunization coverage. Two hundred and sixty-five children, between the age of 9 and 24 months were enlisted for the study, using a practical cluster sampling method. Immunization status of the children, visits to health facilities after 9 months of age and reasons for non-immunization were noted. Missed opportunities for measles immunization was defined as any visit by an eligible child to a health facility, which did not result in his/her vaccination. Measles immunization coverage amongst the study population was 50.61%. The prevalence of missed opportunities for measles immunization was 14.3%. Not offering measles immunization at curative visits (86.9%) was the major reason for missing opportunities. Avoiding all missed opportunities could have increased the measles immunization coverage to 64.9%. PMID- 10798104 TI - Serum TNF-alpha and free radical scavengers in neonatal septicemia. AB - Tumor necrosis factor-alpha (TNF-alpha) and free radicals have been implicated in the pathogenesis of neonatal septicemia and its complications. This case control study was conducted between November 1996 to July 1997 to determine the levels of TNF-alpha and free radical scavengers viz. superoxide dismutase (SOD) and glutathione peroxidase (GPX) in the serum of 30 septic neonates and 20 healthy controls. Patients with neonatal sepsis registered significantly higher levels of TNF-alpha, SOD and GPX in comparison to controls (p < 0.05). The neonates with septic shock had five fold increase in TNF-alpha levels (2262 +/- 605.8 pg/ml) as compared to those without shock (738.8 +/- 728.8 pg/ml). There was no statistically significant difference in levels of antioxidant enzymes between neonates with shock and without shock. The levels of TNF-alpha and antioxidant enzymes were not affected by the type of organism isolated in blood culture. PMID- 10798106 TI - Anesthesia and surgery in pediatric patients with low hemoglobin values. AB - A low preoperative hemoglobin (Hb) concentration creates a dilemma for the anesthetist to proceed or not. The authors prospectively studied the perioperative morbidity of 200 healthy infants and children (age range 3 mon-5 years) with preoperative Hb values > or = 7 g/dl, undergoing cleft lip and palate surgery under general anesthesia. The patients were later categorized according to preoperative Hb level group A: Hb 7-10 g/dl and group B: Hb > 10 g/dl and the results were compared. One or more episodes of hypoxemia (SpO2 < 91%) were recorded in 8 patients in group A and 6 in group B during airway management. All patients had stable cardiovascular parameters except for transient bradycardia during desaturations. Recovery was similar and rapid. There were no differences in perioperative morbidity between the two groups. However, the safety of general anesthesia in infants and children with difficult airways is questionable when the oxygen reserve is reduced in anemia. PMID- 10798105 TI - Is prognostication in congenital diaphragmatic hernia possible without sophisticated investigations? AB - Congenital diaphragmatic hernia is a complex disorder, in which the anatomical defect is only one part of the spectrum of disease. Hypoplasia of lung complicated by pulmonary hypertension and right to left shunting results in serious hypoxemia. Many factors, based on degree of alterations in respiratory physiology and involving analysis of blood gases and acid base systems, have been used in an attempt to prognosticate the outcome. Majority of these investigations are not available in a modest set up like ours. The case records of all 20 patients admitted and operated for congenital diaphragmatic hernia in pediatric surgery unit of Government Medical College Hospital, Jabalpur from 1978 to 1997 were reviewed retrospectively in an attempt to prognosticate without the sophisticated investigations. It was found that even in a very modestly equipped hospital it is possible to prognosticate--to some extent--the outcome in these cases. Major prognosticators found were APGAR score (if child born in hospital), late age of presentation, location of stomach and identification of hernial sac. PMID- 10798107 TI - Perceptions of teachers regarding sex education in National Capital Territory of Delhi. AB - A cross sectional study was conducted to find out the knowledge and attitudes of school teachers regarding sex education. Information was collected from 476 senior secondary school teachers belonging both to the government and public schools, selected randomly in National Capital Territory of Delhi using pre tested close ended questionnaires. A majority of school teachers (73%) were in favour of imparting sex education to school children. Regarding contents of sex education, 90% agreed to the inclusion of reproductive anatomy, physiology including menstruation and birth control measures like condoms and oral pills. However, a majority of school teachers did not want sex education to include topics like abortion, premarital sex and masturbation etc. Fourteen years of age was considered to be the most appropriate for imparting sex education by 28.6% of school teachers. School teachers and doctors were considered by 69.4% and 63.6% of the respondents respectively to be the most appropriate persons for providing sex education. PMID- 10798108 TI - Approaches to sample size calculation in comparative studies. AB - The questions 'what should be the minimum study subjects' and 'how these subjects should be enrolled in the study' are two aspects that are considered at the design stage. Regardless of research question and the study design adopted, these two questions are an integral part of any research proposal. No matter how well a study is conducted and statistically analysed using sophisticated statistical methods/softwares, if the study size is inadequate, concluding statements will have inadequate power and such statements may be misleading and harmful to both target population and the scientific community. There is no substitute/shortcut to an adequate study size as no statistical methods are available to adjust for an inadequate study size. Though a larger sample size will result in higher precision in estimates, however, it may not be desirable in terms of cost, time and the efforts. In medical sciences, majority of research questions pertain to comparing outcome measures in two or more than two groups. Focus of this article is on the number of study subjects required in a study and an attempt is made to review the ingredients required to calculate the sample size in situations commonly encountered by researchers in medical sciences. Formulae for the specific situations are provided with worked out examples. PMID- 10798109 TI - Kala azar. AB - Although visceral leishmaniasis (VL) or Kala-azar caused by Leishmania donovani (LD) is globally, a public health problem and its eradication is of high priority, yet until now, no animal reservoir has been found in India and transmission of Kala-azar occurs from man to man through the recognised vector P. argentipes. The cutaneous or dermal leishmaniasis is caused by L. tropica and this is restricted to Rajasthan where it is zoonotic. Investigation by Sharma et al revealed the presence of zoonotic reservoir in Indian desert gerbils, the rodent burrows of which yielded two species of sandfly. PMID- 10798110 TI - Epidemiology and control of malaria. AB - Malaria is essentially a local and focal disease since its transmission depends greatly on local eco-environmental conditions. In India, the incidence of total malaria cases has been contained to around 2-3 million cases per year. However, with the occurrence of outbreaks in different parts of the country since 1994, mortality due to malaria has increased. The challenges posed by changing epidemiological paradigms of malaria, occurrence of outbreaks, increasing trends of P. falciparum cases, spreading resistance to available anti-malarials and development of resistance of vectors to insecticides are areas for major concern. The National Anti-Malaria Programme has made necessary modifications/adjustments in the malaria control strategy with the adoption of the Global Malaria Control Strategy. PMID- 10798111 TI - Behavioural pediatrics. PMID- 10798112 TI - Substance abuse: a pediatric concern. AB - Use of alcohol, tobacco, and other drugs is a worldwide problem and affects many children and adolescents. The use of opiates, cannabis, tobacco, and alcohol have been well recognized for centuries in India. Abuse of alcohol is widespread in college campuses. In the United States, in 1997, 54.3% of high-school students had used an illicit drug by the time they entered 12th grade. In India the use of drugs and alcohol within the context of religious beliefs and local traditions has been historically documented. Children & adolescents are exposed to alcohol, cannabis products, and tobacco product. Several risk factors and protective factors have been identified. A screening psychosocial history is the most useful tool to identify non-specific indicators of substance abuse. In India the use of pan masala, and home brewed products containing marijuana and alcohol have special significance. Pan masala is shown to have carcinogenic, genotoxic, and clastogenic properties. The pediatrician can play a vital role in the appropriate recognition and early referral. PMID- 10798113 TI - Substance abuse in children and adolescents. AB - Consumption of licit and illicit substances has increased all over the world and the age of initiation of abuse is progressively falling. The common drugs of abuse amongst children and adolescents in India are tobacco and alcohol. Use of illicit drugs like cannabis and heroin have also been reported. A high prevalence of drug use and even intravenous use among street children and working children is a matter of concern. Although initiation to drug use usually occurs during adolescence, the adolescent drug users are seldom seen in various treatment centres. Thus community based programmes are beneficial for prevention and treatment of substance abuse among children and adolescents. PMID- 10798114 TI - Medical evaluation of child abuse. AB - Child abuse is a world-wide problem and has existed in all societies and cultures in various forms since ancient times. Intrafamilial physical and sexual abuse is well recognized in United States and other western countries. In countries like India there is a general lack of sensitivity to the issue among professionals and the lay public alike. The pressing issues of poverty, malnutrition, and infectious diseases take priority on resource allocation. Although some of the methods of child rearing and discipline may be culturally sanctioned and inherent, it does not necessarily mean they are harmless. In this paper the authors present clinical signs and symptoms of physical and sexual abuse which occur in the intrafamilial context and are of direct relevance to the medical practitioner. PMID- 10798115 TI - Interpersonal violence, aggression, and antisocial behaviours in the adolescents. AB - Violence is a growing problem among adolescents all over the world. Exposure to violence can have lasting and pervasive effects on an adolescent's mental and physical health, general well-being, and ability to become a productive adult. Research on adolescent violence in India and Southeast Asia is limited; very little is written in clinical journals. Addressing adolescent violence is currently a low priority for medical practitioners because disease, poverty, and infant maternal health pose more immediate threats to morbidity and mortality in Asia. Physicians, especially in India, have a unique opportunity to take preventative actions now, to stem the tide of morbidity and mortality from gun violence that plagues the United States. Adolescents in Asia are at greatest risk for violence exposure in their homes. Pediatricians who are proactive and educate their patients, families, and the community can help reduce or prevent morbidity and mortality resulting from violence in adolescents. PMID- 10798116 TI - Media violence. AB - For decades, media violence has been viewed as largely a Western problem. New studies indicate that Indian children have increasing access to the media and that media violence will subject them to the same problems as Western children: imitation, desensitization, fear, and inappropriate attitudes about violence and aggression. Solutions exist but will have to be implemented within the next decade to protect Indian children and adolescents from the harmful effects of media violence. PMID- 10798117 TI - Pediatric depression and psychopharmacology. AB - Children can and do have depression which is similar to and continuous with that in adults. It can be diagnosed and treated similarly. Clinical evaluation reveals depression and laboratory tests are not required. It may be comorbid with other psychiatric or physical disorders. It causes considerable distress and impairment and suicide can be one serious sequel. Relapse or recurrences can occur. Etiology of depression is uncertain though hypotheses of psychogenesis or biogenesis exists. Psycho-pharmacological advances have helped in understanding etiology of depression and also in its effective treatment. So far, fluoxetine in the only antidepressant shown to be effective though others are also used. Antidepressants need to be given for one year or more. PMID- 10798118 TI - Melas syndrome. AB - An 11 year old male presented with headache, vomiting and weakness of right side of body. One day after admission he developed right focal seizures. He had 5 previous episodes of stroke, the first at 11 months age. His milestones were normal upto the first episode but subsequent mile stones were delayed. His serum and CSF lactic acids were raised. Muscle biopsy showed ragged red fibres on modified Gomori-trichrome staining. His EEG, CT scan and MRI were normal this time. The child improved spontaneously after 7 days. His recovery time progressively became shorter with each episode of stroke. Maximum time for recovery was noted during first episode and least in current episode. This is the first report of Melas syndrome in Indian literature. PMID- 10798119 TI - Jordans' anomaly. AB - A case of Jordans' Anomaly of leucocytes is reported in a young boy with congenital ichthyosis and hepatosplenomegaly. Cytoplasmic vacuoles were seen in granulocytes, monocytes and lymphocytes of the patient and his father. Serum triglyceride was found elevated in the child but not in the father. Ultrasonogram of the patient's liver showed features suggestive of fatty change, thus pointing to a possible abnormality of systemic triglyceride storage. PMID- 10798120 TI - McCune Albright syndrome and hypophosphatemic rickets. AB - A 4 year-old girl had coxa vara, tibial bowing and hyperpigmented macules. The x ray showed polyostotic fibrous dysplasia and evidence of rickets. Biochemical investigations confirmed hyperphosphaturic hypophosphatemic rickets. The literature has been reviewed. PMID- 10798121 TI - Hemophagocytic lymphohistiocytosis. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare condition in children associated with immunodeficiency, life threatening infections and malignancy. Infection associated hemophagocytosis responds well to appropriate antimicrobioal therapy and rarely to steroids when the infective agent is suspected to be of viral origin. PMID- 10798122 TI - Essential concepts in clinical epidemiology for pediatric practice. AB - Providing the best possible health care requires the integration of individual clinical experience with external clinical information, often derived from published research papers. This article reviews several important concepts in clinical epidemiology that should help the pediatric clinician become a more active and critical reader. Among the basic concepts reviewed are measurement of disease occurrence, relative risk, and screening within the context of the pediatric practice. PMID- 10798123 TI - Validity of causes of infant death by verbal autopsy. AB - The aim of this study was to investigate the suitability of the verbal autopsy technique in identification of cause of death during infancy. The study was carried out in 23 randomly selected clusters with 70 live births each, in districts Cuttack and Koraput of Orissa State. Each death occurrence during the reference period (1992-93) was investigated by the field investigator and cross checked by a medical professional using the same proforma. The probable cause of death in each case was analysed separately by the field investigator and then by the professionally qualified medical person. These were then compared using suitable statistical tests. A total of 179 infant deaths and 16 still births were reported by the field investigator in the area as compared to 183 deaths and 12 still births by professionally trained personnel. There was an agreement for 85.79 percent deaths and 75 percent still births. Disagreement was observed for fever, pneumonia, septicemia and meningitis as causes of deaths. It was more in rural area as compared to urban areas. The sensitivity and specificity of the tool were found to be 85.6 percent and 90.4 percent respectively. The technique is found to be cost effective, time saving and reasonably reliable. After intensive training the lay reporters can be used for recording easily recognizable signs and symptoms of infant deaths and thus help in monitoring infant mortality rate and causes of infant death in a vast country like India so as to achieve "health for all" by 2000 AD. PMID- 10798125 TI - Etiology of chronic diarrhea. AB - With worldwide use of oral rehydration solutions, the treatment of acute diarrhea does not pose much of a problem. However, chronic diarrhea is still harmful, especially for the growth and development status of the children. Between January 1993 to December 1996, patients who suffered from chronic diarrhea for more than one month duration and admitted to Dr. Sami Ulus Children's Hospital were evaluated for epidemiological and etiologic factors. Seventy consecutive patients were evaluated. The mean age was 40.8 months and 52% were males. Malnutrition was detected in 80% of cases. Etiologic factors included celiac disease 30%, cow milk allergy 17%, bacterial and parasitic factors 26%, cystic fibrosis 10% and postinfectious gastroenteritis 10%. Eosinophilic gastroenteritis, chronic nonspecific diarrhea, pseudo-obstruction, neurofibromatosis and inflammatory bowel disease were rarely detected. Celiac disease and cow milk allergy were implicated as the most common causes of chronic diarrhea. The vicious cycle of faulty nutrition, malnutrition and infection and postinfectious enteropathy were also significant factors in the etiology of chronic diarrhea. It may be considered that cow milk protein prick test, sweat test, immunologic tests and mucosal biopsies should be performed for the definite diagnosis of chronic diarrhea. PMID- 10798124 TI - Antioxidant defense systems in newborns undergoing phototherapy. AB - This paper was designed to investigate whether phototherapy is an oxidative stress in newborn infants undergoing phototherapy. A day-light continuous phototherapy was given to jaundiced 20 term and 16 preterm newborns for 72 hours. We measured serum vitamin E and the activities of red blood cell anti-oxidation enzymes (superoxide dismutase, catalase and glutathione peroxidase) before and after 72 h of phototherapy. Serum vitamin E levels were not different before and after 72 h of phototherapy in both preterm and term infants. In several studies, antioxidant enzyme activities have been shown to increase in response to oxidative stresses. In this study, however, the antioxidant enzyme activities in the hemolysate were similar before and at the end of the phototherapy in both preterm and full term. In conclusion, the results of our in vivo study do not confirm the thesis that phototherapy is an oxidative stress in newborn infants. Therefore, phototherapy would preferably seem to be safe and efficient method of treatment for all neonates presenting with hyperbilirubinemia. PMID- 10798126 TI - Effect of Lactobacillus supplementation on immune status of malnourished pre school children. AB - The present study was undertaken to assess the effect of Lactobacillus supplementation on immune response of pre-school children. Thirty malnourished (grade I and grade II) pre-school (3-5 years old) children were selected for the study. These were further categorized into three groups 'A', 'B' and 'C', each group comprising of 10 individuals. Groups 'A' and 'B' were given lactobacillus supplementation 1.0 x 10(8) million viable spores/day/child and 2.0 x 10(8) million of viable spores/day/child for 90 days respectively. Group 'C' served as control. Serum adenosine deaminase (ADA) which is an important immuno enzyme marker for assessing lymphocyte function and proliferation was determined at different intervals. The results revealed that lactobacillus supplementation at level of 1.0 x 10(8) million viable spores/day/child for 15 days along with regular diet significantly improved the ADA levels of pre-school children indicating therapeutic value of lactobacillus as supplementation to malnourished pre-school children. PMID- 10798127 TI - Intestinal helminth infections among school children in Visakhapatnam. AB - A study was carried out between 1993-1994 to determine the prevalence and intensity of intestinal helminth infections among 217 primary school children in Relliveedhi, a slum area in Visakhapatnam. The children were between 7 to 13 years of age and belonged to lower socio-economic status. Stool samples collected were processed by modified formalin-ethyl acetate sedimentation technique to determine ova counts (EPG). One hundred and seventy seven children were infected with one or more of the intestinal parasites viz. Ascaris lumbricoides, Trichuris trichiura and hookworm. The overall prevalence of infection was eighty two per cent. Ascaris lumbricoides was the most common infection with a prevalence of 75% followed by T. trichiura (66%) and hookworm (9%). All infected children received a single dose of albendazole (400 mg/child). Single stool samples were examined over a period of nine months to study reinfection dynamics. PMID- 10798128 TI - Hypertonic saline enema in gastrointestinal ascariasis. AB - Many dangerous surgical complications like intestinal obstruction, acute appendicitis with perforation, ileal perforation in a typhoid patient, Meckel's diverticulitis, disruption of post operative intestinal anastomosis, volvulus, and intussusception are known to occur due to ascariasis, with considerable morbidity and mortality. In this retrospective study of 250 cases of gastrointestinal ascariasis admitted in paediatric surgical wards of Govt. Medical College, Jabalpur (MP), the authors analysed the results of conservative (especially the use of hypertonic saline enema-given just like an ordinary soap water enema but substituting freshly made hypertonic saline in place of soap water) and surgical treatment. The success rate of conservative treatment was 95.6%. Hypertonic saline passes through the incompetent ileo-caecal valve (present in 80% of children) and irritates the worm bolus commonly situated in the terminal ileum, causing it to disintegrate. It also helps to increase the intestinal motility and passage of worms into the colon. The use of hypertonic saline enema is safe and effective in the conservative treatment of gastrointestinal ascariasis. Authors feel that it is the most grossly under utilized part of conservative treatment and deserves to be known and used on wider scale. PMID- 10798129 TI - Zinc levels in women and newborns. AB - Zinc is an important trace element having a definitive role in the metabolism, growth and development and reproduction. During pregnancy the requirements for zinc increase. This study was designed to evaluate the zinc status of normal women, normal pregnant women and their newborn babies. Forty normal adult females, 40 normal pregnant women and their newborn babies were randomly selected and their serum and hair zinc levels were analysed using atomic absorption spectrophotometer. The mean serum and hair zinc levels in normal women were 69.47 +/- 1.4 micrograms/dl and 147.45 +/- 6.12 micrograms/g respectively. The mean serum and hair zinc levels in normal pregnant women were 69.0 +/- 3.22 micrograms/dl and 142.83 +/- 4.39 micrograms/g respectively while the mean serum (cord blood) and hair levels in normal new born babies were 72.77 +/- 5.14 micrograms/dl and 188.36 +/- 4.12 micrograms/g respectively. There was a significant (p < 0.001) decrease in hair zinc levels during pregnancy. There was a significant (p < 0.05) decrease in zinc levels in new born babies when the time interval between the previous delivery and the present delivery was less than 3.4 years. The results of the present study reinforce the need for zinc supplementation during pregnancy especially if the interval between pregnancies is short. PMID- 10798130 TI - Zinc and copper in preterm neonates: relationship with breast milk. AB - The study was conducted to assess copper and zinc levels in neonate's serum, mother's serum, neonate's hair and urine and to ascertain association between them. It is of concern whether zinc and copper deficiency is present at birth and maternal blood and breast milk zinc and copper levels have any effect on this. The study sample included 155 neonates with gestational age 26-41 wks and birth weight 0.550-3.800 kg. Mother's serum, breast milk, neonate's serum, hair, urine samples were analysed for zinc and copper by atomic absorption spectrophotometry. Gestational age was estimated either singly or by combination of date of last normal menses, fetal ultrasonography, and postnatal measures of physical and neurological development by clinical examination and weight by Secca electronic balance. The neonates were classified into term and preterm, small (SGA) and appropriate for gestational age (AGA). Neonates over 37 wks and 2.5 kg served as controls. To assess the dependency, relationship and effectiveness of quantitative predictive variables on the predictions of values, multiple regression analysis was used. Neonates between 26-30 wks gestational age and < 2.5 kg birth weight had significantly low serum zinc and copper. Breast milk zinc was low in mothers delivering preterm and < 2.5 kg neonates. Urinary copper and zinc levels were high in preterm appropriate for gestational age (Pre AGA) than term neonates. Multiple regression analysis revealed that neonate's serum Cu, serum Zn, hair Cu, hair Zn, urine Cu and urine Zn had contribution variability of 49.8%, 51.8%, 49.2%, 16.6%, 52.2% and 68.9%, respectively. The effect of mother's serum, breast milk, and neonate's serum copper and zinc collectively was significant for serum copper (F = 29.59) and hair zinc (F = 32.03). Preterm and low birth weight infants during subsequent growth and development should be supplemented with zinc and copper when on breast feeding. PMID- 10798131 TI - Spinal dysraphism. AB - Spinal dysraphism (SD) is characterized by maldevelopment of neural tube, notochord, mesoderm and cutaneous ectoderm. Incidence of SD is 2-4/1000 live births. One hundred and nineteen patients operated from January 1991-June 1996 at Department of Neurosurgery, All India Institute of Medical Sciences, were studied. Only 21 patients (17.6%) presented when they were less than one year old and 17 patients came in adult age group (> 16 years). Lumbar and lumbosacral region was the commonly involved site in 81 patients (74.7%). Weakness of lower limbs (74%), difficulty in walking (54%), muscle atrophy (41.2%) were the commonest indicators of motor system involvement. Loss of sensation, trophic ulcer, backache were seen in 45, 14, 10 patients respectively. Cutaneous lipoma (26%), hypertrichiosis (20%), dermal sinus (13.4%), midline dimples (7%) were the important cutaneous markers. Foot and limb deformity was seen in 25% cases. Tethering of cord, syringomyelia & split cord malformation were the most common radiological findings. Only 10% of our patients had hydrocephalus that required shunt. Out on 119 cases operated, 43 improved, Twenty had sensory improvement and 18 showed motor improvement. Fifteen patients regained continence. Twelve patients were lost to follow-up. Sixty-seven patients had no change in neurological status, post-operatively. Six cases deteriorated in terms of motor or sensory deficit and one patient lost continence. CSF leak (8%) and wound infection (6%) were the common complications. Six patients required second surgery as T.P. Shunt (4), rotation flap (1), reexploration and duraplasty (1). PMID- 10798132 TI - Use of eutectic mixture of local anesthetics in children. AB - The Eutectic Mixture of Local Anesthetics (EMLA) is a topical application, which has proved to be a useful medication for providing pain relief among children. It is an emulsion containing a 1:1 mixture of lidocaine and prilocaine. The high concentration of the uncharged anesthetic base in the microdroplets of the emulsion ensure effective skin penetration. In the pediatric population EMLA has been shown to be efficacious when it is used prior to venipuncture, cannulation, lumbar puncture, laser treatment of port wine stains, curettage of molluscum contagiosum or vaccination. For several of these indications, the efficacy has been documented by double blind controlled trials, that have used objective and quasi-objective scales for assessing pain relief. The dose of EMLA is between 0.5 to 1 gram, and the cream should be applied half to one hour prior to the procedure. Local side effects are very mild, and the only systemic side effect of importance is the risk of methemoglobinemia in young infants. The literature has conflicting reports about the safety of EMLA in neonates. PMID- 10798133 TI - Latex allergy in children: diagnosis and management. AB - Latex allergy is an increasingly common condition, in both children and health care workers who provide care for them. Subpopulations at particular risk include children with spina bifida, children undergoing multiple surgical procedures, and health care workers in the operating theatre. Chemical additives in latex gloves can cause an irritant or allergic contact dermatitis. Latex proteins are responsible for most of the immediate IgE-mediated hypersensitivity allergic reactions. Symptoms range from rhinitis, conjunctivitis and urticaria to anaphylaxis and death. A latex-directed history is the primary method of identifying latex sensitivity, although both skin and serum testing is available and increasingly accurate. (Latex avoidance should be used in all individuals with a positive skin or blood test or a positive history). The most important preventive measure for patients with or at risk for latex allergy is minimizing direct patient exposure to latex products, most notably latex gloves. Recent operating room studies indicate simple preventive measures can dramatically reduce intraoperative reactions. Preoperative prophylaxis with antihistamines and steroids have not been shown to be necessary or effective. Treatment of an allergic reaction begins with immediate removal of any identified source of latex in direct patient contact. Treatment is similar to anaphylaxis from other causes, and may require the use of epinephrine. Everyone caring for the patient at risk for latex allergy must be involved in making their medical environment safe. PMID- 10798134 TI - Pediatric gastritis and peptic ulcer disease. AB - Inflammation of the gastric and duodenal mucosa is the end result of an imbalance between mucosal defensive and aggressive factors. The degree of inflammation and imbalance between defensive and aggressive factors can then result in varying degrees of gastritis and/or mucosal ulceration. Gastritis and ulcers of the duodenum or stomach can be classified as primary or secondary. The majority of children with chronic gastritis and ulcers in the stomach or duodenum have secondary inflammation or mucosal ulceration. These secondary ulcers generally occur due to a systemic condition like head trauma or overwhelming sepsis, or as sequelae to drug ingestion (i.e. non-steroidal anti-inflammatory agents), but secondary gastroduodenal ulcers can also occur in specific disease conditions such as Zollinger-Ellison syndrome or Crohn's disease. In almost all children with primary duodenal or gastric ulcers mucosal inflammation and, less frequently, ulceration is caused by a spiral shaped, gram-negative, microaerobic rod Helicobacter pylori. Recent epidemiological evidence has linked chronic H. pylori infection with the development of gastric carcinomas. PMID- 10798135 TI - Advances in neonatal surgery. PMID- 10798136 TI - Prenatal diagnosis and assessment of fetal malformations by ultrasonography in India. AB - A multicentre study to assess the status of prenatal diagnosis of fetal malformation in India was conducted. Questionnaires were sent to the teaching hospitals and to centres in the private sector involved with ultrasonography in all the four zones. Data were obtained from 13 centres. Basic level I scans were performed on all pregnant women in 64% centres, whereas level II or targeted scans were performed as routine in 42% centres. Obstetricians performed level II scans in only 35% of cases while rest were performed by radiologists. Malformations of the central nervous system were commonest, accounting for 35-69% of all malformations, followed by genitourinary and gastrointestinal malformations. Malformations of the cardiovascular system were detected in 9.3% as abnormal four chamber view and outlet tract abnormalities, subsequently confirmed on fetal echocardiography. Invasive prenatal diagnosis by fetal blood sampling, chorion villus sampling and amniocentesis for chromosomal analysis of malformed fetuses was performed at 40% centres. At the All India Institute of Medical Sciences, approximately 11,042 ultrasound scans were performed by obstetricians as routine in all pregnant women. A total of 543 malformations were detected accounting for a 4.9% incidence of structural anomalies. A total of 98 cardiovascular malformations were detected in high and low risk pregnant women on fetal echocardiography. No database or registry exists in India to give an actual insight into the problem of congenital malformation and their prenatal diagnosis. It is important to train obstetricians in level I and II ultrasonography, to recognise centres and institutions at national and regional level, and to integrate a network of diagnostic testing. Close liaison between geneticists, radiologists, pediatric surgeons, pediatricians and obstetricians is a must for prenatal diagnosis and management of fetal malformations. PMID- 10798137 TI - Antenatal urologic intervention: current status. AB - The devastation caused by fetal obstructive uropathy is now well known. At the most severe end of the spectrum of obstructive uropathy not only is the developing kidney damaged but the resultant oligohydra-mnios prevents pulmonary development and causes skeletal defects. The most significant changes are noted in patients with posterior urethral values (PUV). The obvious solution to the problem is to either correct or by pass the obstruction prior to the development of permanent changes. Unfortunately, this simple concept is not easy to apply since it raises numerous ethical, legal, economic and technical problems. PMID- 10798138 TI - Esophageal atresia and tracheo-esophageal fistula: a review. AB - The improved survival of neonates with esophageal atresia and tracheo-esophageal fistula reflects the advancement in neonatal care and anaesthesia over the years. Chick embryo studies have given new insights in the embryopathy of esophageal atresia. It is now apparent that the various types of esophageal atresia could be explained due to selective discrepancy in the growth of the 3 folds in the region of tracheo-esophageal separation. The early disturbances in organogenesis which result in esophageal atresia also lead to other associated anomalies, the incidence of which varies from 40 to 55%. These anomalies have an important bearing on the survival outcome. The physiological aspects of esophageal atresia such as esophageal dysmotility and gastro-esophageal reflux are also vital in the long term and proper treatment of the associated defects. The criteria for an ideal esophageal substitute in long gap esophageal atresia have been determined and several options are now available with good results, such as: gastric transposition, colon, gastric tube and small intestine. IN developing countries, however, a high mortality is still attributed to late referrals, low birth weight, hypothermia and chest infection. PMID- 10798139 TI - Gastroschisis and omphalocele. AB - The survival rate of patients with abdominal wall defects has gradually improved with the advances in the investigation and treatment modalities. The present paper reviews the results of various treatment modalities and also analyses the long term results in these patients. A meta-analysis was performed via a medline search of English written clinical studies containing the text words "abdominal wall defects", gastroschisis and 'omphalocele or exomphalos" from 1953 to 1998. The present consensus on operative management of abdominal wall defect is to provide primary closure, if it can be achieved without haemodynamic or respiratory compromise. Patients with primary closure on analysis were found to have better survival rates, reduced risk of sepsis and overall, a shorter hospital stay. However, resumptions of oral feeds, duration of total parenteral nutrition (usually lasting 10-15 days) and ventilatory support required postoperatively did not significantly differ in the primary and silo technique. Long term outcome of these patients is generally good, but they have high incidence of GER (40-50%) for which they should be on regular follow up. PMID- 10798140 TI - Management of anorectal malformation in neonates. AB - Anorectal malformations are one of the commonest anomalies in the new born. Major advances have been made in the last decade in operative techniques to reconstruct this abnormality. The final outcome in these babies is dependent on careful planning and operative intervention in the neonatal period. The purpose of this paper is to discuss the varied presentations of this anomaly, initial assessment and operative management with reference to our own experience. PMID- 10798141 TI - Pre-natal echocardiographic diagnosis and neonatal balloon dilatation of severe valvar pulmonic stenosis. AB - A case is presented below where pre-natal echocardiographic diagnosis of critical pulmonic valvar stenosis was made at 36 weeks of gestation. In view of the severe heart failure, successful balloon valvotomy was performed on day 4 of life. The child was asymptomatic at one month follow-up. PMID- 10798142 TI - Cryptophthalmos. AB - Cryptophthalmos, a very rare congenital anomaly of the eye, is characterised by skin passing continuously from the forehead to the cheek over a malformed eye. It may be isolated or more commonly as a part of Fraser syndrome. Cryptophthalmos is classified into three types: complete, incomplete and abortive. Surgical reconstruction is the modality of treatment available. We present a case of bilateral, complete, isolated cryptophthalmos, who has undergone stage 1 of reconstructive surgery. PMID- 10798144 TI - Unusual presentation of tuberculosis in an infant. PMID- 10798143 TI - Osteopetrosis--improvement of hematologic findings with age. AB - A three months old girl was admitted to the hospital for evaluation of pallor and abdominal distension. There was anemia, thrombocytopenia and extramedullary hematopoiesis. In the long bone roentgenogram medullary areas could not be distinguished and bone densities were increased. The patient was diagnosed for osteopetrosis with these findings and prednisolone treatment was begun and then discontinued since it was not effective. In the follow up, the need for blood transfusions decreased and then disappeared. We present a case which was diagnosed as osteopetrosis and followed up by blood transfusions and of which hematologic findings improved with age. PMID- 10798145 TI - Smoking and children. AB - Tobacco use by children and adolescents is a major health threat. A number of carcinogens and other harmful compounds have been identified in tobacco smoke. The major component, nicotine, is highly addictive. In India, approximately 5500 children and adolescents start using tobacco products daily, some as young as 10 years old. The majority of users have first tried tobacco prior to age 18. Children and adolescents are exposed to the harmful effects of nicotine from smoking or second hand smoke from others; and from use of smokeless tobacco. There is increased prevalence of respiratory disease, ear and sinus infections, asthma, oral disease, and many long-term complications such as cardiovascular disease and cancers due to tobacco use. Prevention and treatment strategies include behavioural approaches and pharmacotherapy. There is an increased urgency especially, for countries like India to address the problem of tobacco use by children and adolescents as the tobacco industry faces legal and public opinion obstacles in Western countries like United States. The medical practitioner can play an important role by implementing the preventive and treatment strategies in his or her practice. PMID- 10798146 TI - National Vitamin A Prophylaxis Programme: need for change in current age strategy. AB - Child survival and Safe Motherhood Programme emphasises on giving vitamin A prophylaxis upto three years of age only, contrary to earlier practice of its administration upto six years of age, based on the assumption of reduction of serious manifestations of vitamin A deficiency three years of age onwards. A cross-sectional study enrolling 1094 children was done to investigate vitamin A deficiency in under six children in urban slums of Nagpur city in Central India in post CSSM scenario. Clinical as well as subclinical (detected by abnormal conjunctival impression cytology) assessment of vitamin A status was performed according to standard procedures, as per WHO recommendations. The overall prevalence of xerophthalmia was 8.7%. Only milder manifestations of xerophthalmia were observed. Significantly higher prevalence of xerophthalmia was observed in more than three years of age. Although nonsignificant, higher prevalence of subclinical vitamin A deficiency was observed in above three years of age group. In view of current age strategy for vitamin A supplementation (< or = 3 years) and observed higher prevalence of clinical and subclinical vitamin A deficiency above three years of age in this study (also endorsed by earlier studies) a call for review of current age strategy for vitamin A supplementation is warranted. PMID- 10798148 TI - Sero diagnosis of tuberculosis in children using two ELISA kits. AB - The diagnosis of childhood tuberculosis is based on circumstantial evidence in the absence of a gold standard in the majority of cases. Sero-diagnosis offers scope for an early diagnosis in a variety of clinical conditions and is simple to perform. A number of mycobacterial antigens have been used for antibody detection assays and several are available as kits in the market. This study was done to evaluate the value of antibody detection kits (ELISA) against the A60 antigen and 38 kDa antigen of Mycobacterium tuberculosis in the diagnosis of childhood tuberculosis at the outpatient department of the Institute of Social Paediatrics, Government Stanley Hospital in collaboration with Tuberculosis Research Centre, Chennai. Thirty five children with pulmonary tuberculosis, 7 with TB lymphadenitis and 22 healthy controls were studied. In addition to routine investigations including gastric lavage for AFB culture, serum antibodies against the A60 and 38 kDa antigens were assayed using commercially available ELISA kits. With A60, IgM serum levels were positive in 74% of pulmonary TB cases, 57% of TB lymphadenitis cases and 50% of controls. A60 IgG was positive in 17% of pulmonary TB, 86% of TB lymphadenitis and 14% of controls. The 38 kDa IgG antibody was positive in 37% of pulmonary and 86% of TB lymphadenitis cases and 27% of controls. Among 10 culture confirmed cases, A60 IgM was positive in 8, A60 IgG in 3 and 38 kDa IgG in 5 patients. The sensitivity of the tests ranged between 29% and 71% and specificity between 50% and 86%. Although the numbers are small, the results suggest that serodiagnosis using the currently available antigens of M. tuberculosis is unlikely to be a confirmatory test for tuberculosis in children. PMID- 10798147 TI - Blood culture and respiratory syncytial virus identification in acute lower respiratory tract infection. AB - Even though the incidence of pneumonia in developed and developing countries is similar, the mortality is five times higher in developing countries. This study aimed to determine the prevalence of bacteremia in children with acute lower respiratory tract infection (LRTI) and relative contribution of respiratory syncytial virus (RSV). One hundred and one children under five years of age who attended a primary care level clinic with diagnosis of acute LRTI, were enrolled. Diagnosis and management of pneumonia were done according to the WHO guidelines. Two blood cultures were drawn at the time of admission. A nasopharyngeal sample was taken for detection of RSV by indirect immunofluorescence. Blood cultures were positive for pathogenic bacteria (Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus) in three patients. The detection for RSV was positive in 24 patients (23.7%). The clinical and radiographic presentations were not significantly different between patients with and without RSV (p > 0.05). RSV is a common cause of LRTI in children younger than five years old. Blood cultures are not commonly positive in outpatients with acute LRTI. The practice of obtaining blood cultures in primary and secondary care clinics is not useful to guide the treatment of patients with community-acquired pneumonia. PMID- 10798149 TI - Mid-arm circumference for detecting malnutrition during infancy. AB - Mid-arm circumference (MAC) has been extensively used at 1-5 years of age as an age-independent indicator of protein-energy malnutrition (PEM). Its role below 12 months of age has not been studied adequately and a search for a clinically reliable cut-off point has not been systematically done. This study was planned to establish the age-independent character of MAC within the 6-12 month age group and identify a cut-off point demonstrating the maximum sensitivity and specificity in detection of PEM. Two hundred and forty malnourished infants [< 80% weight for age (W/A) of 50th percentile of NCHS standard] were included with even distribution into 6 subgroups; 240 healthy infants were taken as controls. MAC within the age of 6-12 months was found to be age independent with an overall increase of 5.1%. By using various cut-off points a value of < or = 12.5 cm of MAC proved to be having maximum sensitivity (92.1%) and specificity (88.3%). MAC is, therefore, a simple, age independent, informative measurement for early assessment of PEM by using single cut-off point < or = 12.5 cm, and can serve as a screening test for targeting infants for meaningful utilization of nutritional programmes. Since nearly all MAC tapes in current use already have < or = 12.5 cm colour coded cut-off, no new tapes are necessary. PMID- 10798150 TI - Hypophosphatemic rickets: easy to diagnose, difficult to treat. AB - Hypophosphatemic rickets (HR) has generated a lot of interest in recent times. There is need to recognize this disorder and differentiate it from the more common nutritional rickets because the therapy is different. It is also important to emphasize that a detailed clinical examination with pedigree analysis and easily available biochemical tests are adequate to establish the diagnosis in most cases. This report presents three families with hypophosphatemic rickets. Interestingly, many of these patients had a mixed picture of HR and nutritional rickets. Their important features are described with special emphasis on early initiation of treatment with oral phosphate and stringent monitoring of renal functions to prevent development of irreversible renal insufficiency. PMID- 10798151 TI - Manpower utilization in pulse polio immunization. AB - The present study was conducted in Union Territory of Chandigarh to find out the utilization pattern of manpower engaged in the implementation of Pulse Polio Campaign (PPI) on 7th December, 1997 with the objective to achieve a coverage close to 100% and thereby, eradicating poliomyelitis. Over 100,000 under-five children were expected to participate on PPI day. Out of 240 polio centres established in the Union Territory, 16 were selected by stratified random sampling covering 14,858 children. The observing teams recorded the information about the children and presence of staff members throughout the day between 8.00 a.m. and 5.00 p.m. It was observed that nearly half of the children received the polio drops within the first three hours while only a little over 5% visited polio centres in the last two hours. The average number of manpower varied between 5 and 6 throughout the day (more than the recommended four). The staff posted at urban, rural and slum centres did not visit the houses except for the few volunteers in slums. Unimmunized children should be identified by the staff in last two hours when the load at polio centres is extremely low. Adequate utilization of the full potential of the manpower will help in enhancing the PPI coverage close to 100%. PMID- 10798152 TI - Urolithiasis: a tribal scenario. AB - Urolithiasis is quite prevalent in Satpura belt of Central India. Forty five children with urolithiasis were studied in Jawahar Medical Foundations Hospital, Dhule, Maharashtra over a period of three years. The mean age was 7.1 +/- 3.2 (range 18 months-15 years). The chief presenting complaints were dysuria and lower abdominal pain. The physical, laboratory and radiographic findings were evaluated. Calculi were located in the bladder (80%), urethra (9%), kidney (6.6%) and ureter (4.4%). Calculi were mainly composed of calcium oxalate (65.7%) and calcium phosphate (34.3%). The predominant urinary tract stones in the tribal Satpura belt are bladder stones and may be related to poor nutrition, low socioeconomic status and consumption of bajra (millet) as staple food as found in this region (93.5%). PMID- 10798153 TI - Intensive chemotherapy in children with stage IV neuroblastoma. AB - A retrospective analysis of effectiveness of sequential chemotherapy with cyclophosphamide, doxorubicin, cisplatin and etoposide in children with stage IV neuroblastoma was undertaken. Study group included 17 children of mores than one year old with median age of 3 years (range 18 months to 7 years). Fourteen were males and three females. Sites of primary tumor were abdomen in 12 patients, pelvis in 3, paravertebral in 1 and unknown in 1. Metastatic sites included bone marrow (88%), bone (82%), orbit (29.4%) and lymph node (11.7%). One patient had brain parenchymal disease and another had cerebrospinal fluid positivity for malignant cells. Fifteen of the 17 patients had major response with chemotherapy (complete response in two and partial response in 13). Ten of the 15 patients completed four courses of chemotherapy and five patients progressed while on chemotherapy and died. Only two of the ten patients, who had four courses chemotherapy are alive after 2 years. Hence the 2-year survival in this series is 11.7%. There was no toxic death in this study. PMID- 10798154 TI - Acute lower respiratory tract infection: the forgotten pandemic. PMID- 10798156 TI - Recurrent lower respiratory tract infections in children. AB - Recurrent lower respiratory tract infections in children are caused by a deficiency in the host defense or an underlying pulmonary disorder which may result from structural or functional alterations. Aspiration syndromes, congenital defects, airway clearance disorders and immune deficiency are the main categories. Careful evaluation with good history and examination provide important leads to limit the radiological and laboratory work up to the most relevant investigations. Some of the structural abnormalities are amenable to cure; however, secondary events need careful management of the underlying lung disorder to give the best quality of life to these children. PMID- 10798155 TI - Cognitive and behavioural concerns in epileptic children. AB - Cognitive performance in an epileptic child has been a difficult issue to predict in day-to-day clinical practice. Several observations made in early and later part of this century do not provide uniform and convincing answer to this issue. Recent trends in research however, have identified certain variables that are shown to be associated with cognitive decline in epileptic children. Together with associated behavioural problems, the resultant school difficulty is the essence of this concern for the parents. The variables related to cognitive deterioration as identified by several studies include underlying brain pathology (symptomatic epilepsy), early age of onset of seizure, severity and intractability of seizure, repeated head trauma, an episode of status epilepticus, presence of interictal subclinical EEG discharge, adverse psychosocial factor and antiepileptic drug (AED). Association of these variables in a given case cannot only predict adverse cognition outcome but also a preventive management package can be planned aiming at avoiding or minimizing these high risk variables. PMID- 10798157 TI - Pulmonary manifestations of pediatric HIV infection. AB - Vertically acquired HIV infection is becoming increasingly common in India. The main clinical manifestations of HIV in childhood are growth failure, lymphadenopathy, chronic cough and fever, recurrent pulmonary infections, and persistent diarrhoea. Pulmonary disease is the major cause of morbidity and mortality in pediatric AIDS, manifesting itself in more than 80% of cases. The most common causes are Pneumocystis carinii pneumonia (PCP), lymphocytic interstitial pneumonitis (LIP), recurrent bacterial infections which include bacterial pneumonia and tuberculosis. The commonest AIDS diagnosis in infancy is PCP, presenting in infancy with tachypnea, hypoxia, and bilateral opacification on chest-X-ray (CXR). Treatment is with cotrimoxazole. LIP presents with bilateral reticulonodular shadows on CXR. It may be asymptomatic in the earlier stages, but children develop recurrent bacterial super infections, and can progress to bronchiectasis. LIP is a good prognostic sign in children with HIV infection in comparison to PCP. HIV should be considered in children with recurrent bacterial pneumonia, particularly with a prolonged or atypical course, or a recurrence after standard treatment. Pulmonary TB is common in children with HIV, but little data is available to guide treatment decisions. Much can be done to prevent PCP and bacterial infections with cotrimoxazole prophylaxis and appropriate immunisations, which may reduce hospital admissions and health care costs. PMID- 10798158 TI - Pulmonary function testing in office practice. AB - With the wide availability of simple to operate pulmonary function equipment, these tests can now be performed in the office or clinic setting. The use of pulmonary function tests in the assessment of patients with pulmonary diseases has thus expanded. These simple tests rely on data generated from a forced expiratory vital capacity maneuver which can be performed reliably by most children over the age of 6 or 7 years. Pulmonary function tests provide both diagnostic and prognostic information and help in the management of a number of respiratory diseases. The number of tests available vary with the level of sophistication of the pulmonary function laboratory; in most situations simple spirometry is adequate. Several new and inexpensive peak flow meters are now available for home use; these are very useful for home monitoring of asthma patients. Physicians caring for patients with asthma or other chronic respiratory diseases should be encouraged to use these tests routinely in the management of these children. PMID- 10798159 TI - Role of imaging in pediatric respiratory diseases. AB - Both medical and surgical diseases commonly affect the pediatric respiratory system. Imaging has a crucial role to play in their management. Plain chest radiograph is still the single most important investigation. However, at times further workup is essential and then CT, MR imaging and other modalities provide detailed findings. Pediatric radiologist should be aware of the advantages and the limitations of each imaging technique so as to be able to use these judiciously, without duplicating information and at the minimum cost and radiation to the patient. PMID- 10798160 TI - Diagnosis of cystic fibrosis: Indian perspective. AB - Cystic fibrosis (CF) is one of the common life limiting inherited diseases in Caucasian population. Recent reports suggest that the diagnosis of cystic fibrosis in Indian children is missed or delayed due to low index of suspicion. The diagnosis of cystic fibrosis is suspected by the typical clinical features and should be confirmed by doing sweat chloride estimation. If sweat test is not available, ancillary tests including blood electrolyte and acid base balance, airway microbiology, tests to identify pancreatic insufficiency and semen analysis for obstructive azoospermia in post pubertal boys should be carried out. Positive results of these tests make the suspicion very strong. A strongly suspected case should be treated as cystic fibrosis, but for giving a diagnosis of CF, sweat test should be done from the nearest centre where it is available. In the presence of typical clinical features with borderline sweat chloride values sweat test should be repeated 2-3 times and the child should be investigated for alternative diagnosis. In the absence of alternative diagnosis with consistently high or borderline sweat chloride values an attempt should be made to get tests for mutations. PMID- 10798161 TI - Pneumonia due to unusual organisms in children. AB - Generally antimicrobials for treatment of pneumonia are chosen to target the usual bacterial etiological agents. Such regimens are unable to cure patients of pneumonia caused by 'unusual organisms' mycoplasma, chlamydia, Pneumocystis carinii and Legionella pneumophilus). Thus, there is a need to anticipate their presence in appropriate cases and to plan the initial antimicrobial therapy accordingly. Studies in Europe as well as India have shown that such infections form a fairly substantial percentage of community acquired pneumonia in children. Mycoplasma pneumoniae and Chlamydia pneumoniae are common in school age children while Chlamydia trachomatis occurs in early infancy. Pneumocystis carinii is an important pathogen in immunocompromised children. Routine laboratory tests and radiological features are not specific enough to give accurate diagnosis of these infections for which one has to depend on sophisticated culture techniques, immunological tests for the antigens or antibodies and polymerase chain reaction. Mycoplasma, chlamydia and legionella infections respond to macrolide antibiotics and for pneumocystis infections, trimethoprim-sulfamethaxozole or pentamidine is the drug of choice. Overall prognosis with appropriate treatment is good except for P. carinii infection in immunocompromised host which carries a high mortality and recurrence rate. PMID- 10798162 TI - Structural chromosomal anomaly in mental retardation. AB - This article reports the structural chromosomal anomaly in three patients with mental retardation: (i) Proband was a five year old girl with reciprocal retardation (1; 2) (p32; q11) (ii) Proband, female of 14 years. Her karyotype showed translocation (1; 3) (q42; q13). The translocations were de novo in origin (iii) Proband showed variant 13 as the giant satellite over its short arm, and this was paternal in origin. Proband, eighteen months old male child had microcephaly and seizures. These two features may be because of autosomal recessive condition. This report emphasises the need for kayotyping to provide a clear cut diagnosis and appropriate counselling. PMID- 10798163 TI - Congenital spinal lipomas--tumor versus hamartoma. AB - Congenital spinal lipomas as a form of dysraphism are frequently reported in literature. A few studies have highlighted their histologic features and found them different from lipomas in other parts of the body. In a review of eight cases of spinal lipomas, we found heterotopic tissue in seven. Neural tissue, muscle, bone and cartilage were frequently encountered. The presence of meningothelial cells in one case was an unusual finding. These findings support the concept of their hamartomatous origin. Malignancy, occurring later in life in one of the heterotopic elements is known, though uncommon, and is one of the reasons, besides neurologic deficit, for long term follow-up. PMID- 10798164 TI - Megacystis microcolon intestinal hypoperistalsis syndrome. AB - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a rare cause of intestinal obstruction mainly affecting female neonates. We present a case of a newborn female infant with a history of abdominal distension, bilious vomiting and decreased urine output. Barium enema showed a microcolon. Patient died soon after admission and the autopsy revealed a shortened bowel, a microcolon with abundant ganglion cells in the myenteric plexus, and an enlarged urinary bladder. An interesting finding in this case was the presence of enlarged nerve bundles containing several large ganglion cells on the lateral wall of the cervix. The salient clinical and autopsy findings in this case are presented. PMID- 10798165 TI - Bilateral congenital trigger thumb: role of heredity. AB - Bilateral congenital trigger thumb is a relatively uncommon condition, with few well documented cases in literature. Lack of awareness and mode of presentation creates diagnostic confusion to a majority of physicians who are not familiar with this condition. There is also controversy surrounding its aetiology, natural course, ideal treatment and the age for intervention. We report the case of bilateral congenital trigger thumb in a mother and child, review the literature and discuss the possible hereditary influence which is still less known and reported. PMID- 10798166 TI - Publication concerning an article single CT (ring) lesion in epilepsy patients: a new observation by Garg et al, Indian Journal of Pediatrics, January-February, 1999, Vol. 66, No. 1, pages 155-157. PMID- 10798167 TI - [National education physician's strike]. PMID- 10798168 TI - [Organized screening of colorectal cancer in France. System of care and professional logistics]. AB - The preservation or the amelioration of the health of individuals and populations does not only depend on the activity of the medical profession. It largely depends on public health policies. Screening for cancers illustrates how difficult it is to organise an action of public health within a health care system of liberal tradition in France. Through the example of screening for colorectal cancer, this article proposes to identify the mechanisms by which the system of health care determines the methods of organisation of mass cancer screenings. Through interviews carried out among the organisers of five colorectal cancer screening campaigns, we identified three models on which French screening campaigns are based: an administrative model, a medical model and a mixed model. Each of these models represents a different adaptation of the organisation of mass screening to the health care system in place in France. The interests of the concerned professional groups confront each other and the collective interest. It seems that to integrate such an approach to public health in a system dominated by payment per act is doomed to fail. Today, France can no longer avoid a modification of its health care system and of its nomenclature if it wants to improve the feasibility of cancer screening programmes and increase their chance of success. PMID- 10798169 TI - [European public health professionals express ten priorities for the European Union...to be followed]. AB - Even if the European Union acquired explicit competencies in public health with the Maastricht and Amsterdam Treaties (articles 129 and 152), public health professionals still have not had their word in the definition of public health priorities. Yet it is they, whatever their mission, who must take into consideration the new constraints imposed by Community directives. The French Society for Public Health (FSPH) took the initiative of running a project, financed by the European Commission, aiming to shed light on some of the public health problems considered priority in the 15 member states, and to provide suggestions for facing them. The FSPH adopted a resolutely participative and pragmatic process. At each step (definition of priorities and compiling arguments), the intention of the SFPH was more to allow different, even diverging, points of view to be expressed, than to aim for a hypothetical representativeness. The undertaken themes are the social gradients in health, alcohol, illicit drugs, tobacco, surveillance of health issues, quality of care, older persons, mental health, the environment, nutrition and food security. This work marks the wish of the FSPH for international openness toward Europe. The FSPH hopes that this work becomes a platform for the development of a reinforced dialogue between public health professionals and European decision makers. PMID- 10798170 TI - [Use of the admission service and pediatric emergency service of a university hospital center by adolescents (12-19 years): utilization and messages]. AB - A retrospective survey concerning all patients ages 12-19 who had registered themselves at the admitting service of the paediatric emergency unit in the university hospital centre of Amiens was carried out in 1995. 2812 adolescents were received (18% of total admissions). The male/female sex-ratio was 1 to 3. The average age was 13.2 years old. Hospitalisations were more frequent in the spring and were predominantly on Mondays (17%). More than 2/3 of the adolescents were accompanied by their parents and 20% were referred by their doctors or by the SAMU (mobile emergency medical service). Two percent arrive alone. Traumatic pathology is the main reason for admission (1960 patients). Organic pathologies represent 24% of admissions and are essentially digestive pathologies (40%). "Suffering" adolescents (defined as those requiring an intervention with a psychotherapist as a matter of priority) only represent 6% of visits to the emergency unit, of which 2/3 correspond to suicide attempts. 24% of adolescents are hospitalised at the university hospital centre of Amiens within the paediatric services or specialised services (ENT, stomatology, etc.). PMID- 10798171 TI - [Contribution of a combined approach in an analysis of training needs in two health regions in Morocco]. AB - A study using a combination of three techniques of qualitative research (nominal group, focus group, interview) was carried out in 1996 among 72 health professionals that occupy a position of responsibility at the level of the wilayas of Fes and Marrakech (Morocco). For the entirety of the study, 14 meetings were held with experts (administrators, nurses, doctors and delegates). OBJECTIVE: To study the advantages and limitations of a combinatory approach that triangulates three methods of qualitative research to identify and analyze training needs of administrators, nurses, doctors and delegates that hold a position of responsibility at the level of the wilayas of Fes and Marrakech. RESULTS: The focus group allowed for the establishment of lists of problems encountered that were susceptible to being resolved through a training intervention: 1) Planning methods are not used and needs analysis of resources remains arbitrary; 2) Methods of management are not mastered by the people holding the responsible positions; 3) The management of human resources is centralized and there is no policy of personnel motivation. The main accompanying measures proposed by the participants were: 1) to decentralize the process of planning and to train the responsible people in the methods and techniques of planning; 2) to introduce new methods of health services administration. Training themes by order of priority were: health planning, management, regulations and legal instruments, management of human and material resources. DISCUSSION: The combinatory approach allowed for a list to be established of the training needs of health professionals. It also provided an instantaneous diagnosis of the organisations and programmes for which they are responsible. But this triangulation of techniques has several limitations related to the researchers' interpretation of these needs, to their socio-cultural system of reference and to their interests that often don't coincide with those of the experts in the field. CONCLUSION: This study allowed for the development of an outline that puts the role of experts back in the planning process of a training project and combines the needs of experts with institutional needs in order to reach a better adequacy of the triad of needs-demand-responses. PMID- 10798172 TI - [General physicians' viewpoint on the evaluation of health care quality in the hospital]. AB - The principal objective of this work was to determine whether the evaluation of general practitioners' satisfaction vis-a-vis a health care establishment impacted that establishment to improve the quality of the services it provides. Ten doctors were randomly selected and 100% agreed to respond to a semi-directive interview. Two independent doctors analysed each corpus and identified 15 themes, of which 5 systematically came up: the organisation of the care of patients, the availability of hospital doctors, the transmission of information, the speed of emergency admissions, and the patient-hospital doctor relationship. The opinion of private practice doctors usefully complements the evaluation of patient satisfaction used to identify the weaknesses of an establishment. It is proposed to include this evaluation in the procedure of continued improvement of the quality of care within establishments. PMID- 10798173 TI - [Analysis of the management of patients aged 75 and older in an admitting and emergency service of a large hospital]. AB - A prospective survey carried out over two periods (35 days total) presented a typology of people aged 75 and older entering the admitting and emergency unit of a university hospital center, and underlined the specifics of the care they received. These patients represent 27.8% of patients received in the unit. The women (60.9%) live preferentially alone at home or in group establishment. The majority of admissions are the result of a referral from the patients' doctors. Tuesday remains the busiest day for admissions (19.7% of total admissions). 10.9% of patients arrive alone and without health records. The specialist is solicited for 41.3% of older people. Their autonomy level (measured with the AGGI R grid) is evaluated during the emergency visit. An analysis of pathological profiles shows a preponderance of acute problems with risk of degenerative problems (54.9%). Social problems (8.2% of patients) particularly affect the group with multiple pathologies. The choice of placement is more often in specialty services (61.3%) than geriatric services (18.7%). A return to ones previous home is more rarely re-envisaged. The placement obtained corresponds to the desired placement in 68% of cases. Lack of beds is the main cause in cases where the desired placement is not achieved. While the care of older people admitted for "specialised" reasons poses little problem many pathologies including "social" issues, and are admitted for "general" reasons) is a very different issue. PMID- 10798174 TI - [Work schedule management in the health institutions of Abomey-Calavi (Benin)]. AB - Work time management of the health personnel of the health institutions in the sub-prefecture of Abomey-Calavi (Benin) This study was carried out with a sample size of 130, comprising health personnel, patients, members of management committees and twelve health institutions of the subprefecture of Abomey-Calavi. The survey was carried out through observation, one-on-one interviews and consultation of documents. The results reveal that work time is poorly managed in almost all of the programmes surveyed. There exist long waiting lines in almost all the health programmes. On average, the traffic report per service is 82 and 10 out of 12 of the programmes investigated do not have an agenda of activities. PMID- 10798175 TI - [Health status and its effects on the work of national education nurses]. AB - This study presents a pedagogical work carried out with a group of 13 school nurses of the Academy of Rennes, as part of a course in survey methodology. A descriptive survey aiming to achieve a better understanding of experienced health status of nurses, and of its repercussions on the execution of their missions, was implemented during the course, as a result of the demand and the expertise of the participants. Overall, nurses report to be in good health. However, one can not underestimate the existence of a group at risk (5% of the sample) that often suffers somatic troubles accompanied by sleep disorders and anxiety, as these risks concern primarily permanent staff nurses working in boarding establishments, with half of them falling into this category. A demand for regular monitoring of their health was clearly expressed through the study. This may be the object of a subsequent study by an academic team. PMID- 10798176 TI - [Evaluating the coding reference system for morbidity in general medicine]. AB - Many coding systems for morbidity seen in general medicine are currently available in France. This article proposes an approach to evaluating them. Firstly, it is proposed to specify the conceptual framework within which one includes the classificatory approach to health troubles in general medicine. This assumes that the notions of primary care, pathology and diagnosis are clarified in the light of the practices of general practitioners. Secondly, it is proposed to specify the framework of analysis within which one is situated in order to reach these systems of reference that all correspond to classificatory systems. For this, one can examine several dimensions: the purpose of the system, its criteria and main lines of classification, its method of construction, its underlying conceptual model, the type of collection required and the efficiency of data used. Once the conceptual framework is fixed and the framework of analysis chosen, it is possible to evaluate these classifications without ambiguity. PMID- 10798177 TI - [General practice patterns in hepatitis C virus mass screening in the Lyons region]. AB - A study on the screening practices for hepatitis C was carried out among a sample of 786 general practitioners in the Lyon region. The sample comprised three groups: 272 doctors who had participated in a study of this screening, 242 doctors who had not responded to the request for participation, and 272 doctors who had refused to participate. Overall, 88.5% of the doctors surveyed had prescribed at least one HCV serology in the last 12 months, 82% had at least one HCV positive patient and of them, 59.3% had partly or completely screened these patients. The only differences between the three groups are related either to their belonging to an HIV or drug addict care network, or to a particular medical practice (study of allergies, sexology, nutrition, etc.). In fact, the first prescribe more among groups at risk and the second prescribe less and have fewer HCV positive patients. PMID- 10798178 TI - [Participation factors in a occupational health colorectal cancer screening program]. AB - A colorectal cancer screening campaign by Hemoccult test was carried out from January 1993 to December 1994 in collaboration with the company doctors of employees ages 45 and older in the companies of the Lot department of France. Of the 1311 employees to whom the test was offered, 811 actually had the test done, representing a rate of participation of 61.9%. Participation varied from 48.1% to 72.7% depending on the company doctor, and was higher for large companies. Managers participated less than other employees. People who never visit a dentist, who had not seen their doctor for over a year or who never give blood participated less than others. Thus, even though company doctors can play a true role by favouring the participation of general employees, their action is limited by the weak participation of people who already have little contact with the health care system. PMID- 10798179 TI - [Fundamentals of pharmacotherapy and pharmacokinetic principles]. PMID- 10798180 TI - [Effect of genetic factors and illnesses on drug efficacy]. PMID- 10798181 TI - [Drug interactions: new mechanisms and clinical relevance]. PMID- 10798182 TI - [Quality assessment of therapeutic clinical trials]. PMID- 10798183 TI - [Pharmacoeconomy. Aspects of drug efficacy evaluation]. PMID- 10798184 TI - [Role of endothelium in the etiology and therapy of atherosclerosis]. PMID- 10798185 TI - [Anaphylaxis and allergy. Recommendations for emergency treatment]. PMID- 10798186 TI - [Sixteen year old girl with acute liver failure after tonsillectomy]. PMID- 10798187 TI - [Pair of siblings of Italian ethnicity with hyperferritinemia and cataract]. PMID- 10798188 TI - [Intracoronary brachytherapy]. PMID- 10798189 TI - [Adjuvant chemotherapy for colorectal carcinoma]. PMID- 10798190 TI - [Indication for gastric lavage]. PMID- 10798191 TI - [Migraine: clinical aspects, diagnosis and pharmacotherapy]. PMID- 10798192 TI - [Comments on articles by K.H. Gebhardt, "What has been proven in homeopathy?" and K. Linde, "Are there evidence-based therapies in homeopathy?"]. PMID- 10798193 TI - [Comments on article of K.H. Gebhardt, "What has been proven in homeopathy?"]. PMID- 10798194 TI - The unicellular green alga Chlamydomonas reinhardtii as an experimental system to study chloroplast RNA metabolism. AB - Chloroplasts are typical organelles of photoautotrophic eukaryotic cells which drive a variety of functions, including photosynthesis. For many years the unicellular green alga Chlamydomonas reinhardtii has served as an experimental organism for studying photosynthetic processes. The recent development of molecular tools for this organism together with efficient methods of genetic analysis and the availability of many photosynthesis mutants has now made this alga a powerful model system for the analysis of chloroplast biogenesis. For example, techniques have been developed to transfer recombinant DNA into both the nuclear and the chloroplast genome. This allows both complementation tests and analyses of gene functions in vivo. Moreover, site-specific DNA recombinations in the chloroplast allow targeted gene disruption experiments which enable a "reverse genetics" to be performed. The potential of the algal system for the study of chloroplast biogenesis is illustrated in this review by the description of regulatory systems of gene expression involved in organelle biogenesis. One example concerns the regulation of trans-splicing of chloroplast mRNAs, a process which is controlled by both multiple nuclear- and chloroplast-encoded factors. The second example involves the stabilization of chloroplast mRNAs. The available data lead us predict distinct RNA elements, which interact with trans-acting factors to protect the RNA against nucleolytic attacks. PMID- 10798195 TI - Biological information transfer beyond the genetic code: the sugar code. AB - In the era of genetic engineering, cloning, and genome sequencing the focus of research on the genetic code has received an even further accentuation in the public eye. In attempting, however, to understand intra- and intercellular recognition processes comprehensively, the two biochemical dimensions established by nucleic acids and proteins are not sufficient to satisfactorily explain all molecular events in, for example, cell adhesion or routing. The consideration of further code systems is essential to bridge this gap. A third biochemical alphabet forming code words with an information storage capacity second to no other substance class in rather small units (words, sentences) is established by monosaccharides (letters). As hardware oligosaccharides surpass peptides by more than seven orders of magnitude in the theoretical ability to build isomers, when the total of conceivable hexamers is calculated. In addition to the sequence complexity, the use of magnetic resonance spectroscopy and molecular modeling has been instrumental in discovering that even small glycans can often reside in not only one but several distinct low-energy conformations (keys). Intriguingly, conformers can display notably different capacities to fit snugly into the binding site of nonhomologous receptors (locks). This process, experimentally verified for two classes of lectins, is termed "differential conformer selection." It adds potential for shifts of the conformer equilibrium to modulate ligand properties dynamically and reversibly to the well-known changes in sequence (including anomeric positioning and linkage points) and in pattern of substitution, for example, by sulfation. In the intimate interplay with sugar receptors (lectins, enzymes, and antibodies) the message of coding units of the sugar code is deciphered. Their recognition will trigger postbinding signaling and the intended biological response. Knowledge about the driving forces for the molecular rendezvous, i.e., contributions of bidentate or cooperative hydrogen bonds, dispersion forces, stacking, and solvent rearrangement, will enable the design of high-affinity ligands or mimetics thereof. They embody clinical applications reaching from receptor localization in diagnostic pathology to cell type-selective targeting of drugs and inhibition of undesired cell adhesion in bacterial/viral infections, inflammation, or metastasis. PMID- 10798196 TI - Is the newly described Vietnamese bovid Pseudonovibos spiralis a chamois (genus Rupicapra)? AB - The "Linh Duong" (Pseudonovibos spiralis), a newly described Vietnamese bovid, has been shown to be related to the Caprinae on the basis of the mitochondrial cytochrome b gene sequencing from horn fragments. To integrate the greatest available biodiversity we performed a new phylogenetic analysis including all genera of Caprinae. Unexpectedly, Pseudonovibos is found to be robustly associated with Rupicapra, and closer to the Alpine than to the Pyrenean chamois. Several clues led us to interpret this result as a carry-over DNA contamination of Pseudonovibos by Rupicapra. PMID- 10798197 TI - The role of neurosecretory neurons in the pars intercerebralis and pars lateralis in reproductive diapause of the blowfly, Protophormia terraenovae. AB - Microlesions of the brain were made to examine the role of neurosecretory neurons in the pars intercerebralis (PI) and pars lateralis (PL) in the induction of reproductive diapause of the female blowfly Protophormia terraenovae. Under both diapause-inducing (LD 12:12, 20 degrees C) and diapause-averting conditions (LD 18:6, 25 degrees C), the ovaries invariably failed to develop when the PI was removed. When the PL was removed bilaterally, the ovaries developed in most of the females, irrespective of the rearing conditions. Removal of the PL prevented females from entering reproductive diapause. These results show that certain neurosecretory neurons in the PI are necessary for vitellogenesis, and that the PL contains inhibitory neurons which suppress vitellogenesis during reproductive diapause. PMID- 10798198 TI - The use of marine molluskan shells for Roman glass and local raw glass production in the Eifel area (western Germany). AB - Relatively high strontium concentrations and their isotopic composition in Roman glass of the Imperial period indicate the general use of shells as carbonate raw material. Lead, iron, and barium concentrations and lead isotopes of glass of the late fourth century from Hambach and Gellep (western Germany) are conformable with the use of glass sand from Eifel rivers. Each of three pairs of six simultaneously operating Roman glasshouses in the Hambach area made its own raw glass from different quartz sands. PMID- 10798199 TI - Polarization-sensitive interneurons in the optic lobe of the desert ant Cataglyphis bicolor. AB - Desert ants, Cataglyphis bicolor (Hymenoptera), navigate by using compass information provided by skylight polarization. In this study, electrophysiological recordings were made from polarization-sensitive interneurons (POL-neurons) in the optic lobe of Cataglyphis. The POL-neurons exhibit a characteristic polarization opponency. They receive monochromatic input from the UV receptors of the specialized dorsal rim area of the compound eye. Both polarization opponency and monochromacy are features also found in the POL neurons of crickets (Orthoptera). PMID- 10798200 TI - Ice shelf microbial ecosystems in the high arctic and implications for life on snowball earth. AB - The Ward Hunt Ice Shelf (83 degrees N, 74 degrees W) is the largest remaining section of thick (> 10 m) land-fast sea ice along the northern coastline of Ellesmere Island, Canada. Extensive meltwater lakes and streams occur on the surface of the ice and are colonized by photosynthetic microbial mat communities. This High Arctic cryo-ecosystem is similar in several of its physical, biological and geochemical features to the McMurdo Ice Shelf in Antarctica. The ice-mats in both polar regions are dominated by filamentous cyanobacteria but also contain diatoms, chlorophytes, flagellates, ciliates, nematodes, tardigrades and rotifers. The luxuriant Ward Hunt consortia also contain high concentrations (10(7)-10(8) cm-2) of viruses and heterotrophic bacteria. During periods of extensive ice cover, such as glaciations during the Proterozoic, cryotolerant mats of the type now found in these polar ice shelf ecosystems would have provided refugia for the survival, growth and evolution of a variety of organisms, including multicellular eukaryotes. PMID- 10798201 TI - Electric signals in the social behavior of sympatric elephantfish (Mormyridae, Teleostei) from the upper Zambezi River. AB - Electrocommunication in mormyrid fish from African freshwaters is a challenging research field in neuroethology (Turner et al. 1999). However, virtually nothing is known about electrocommunication within natural mormyrid populations involving sympatric, syntopic species. Here we report on the nocturnal activities and electrocommunication among three syntopic species in a spacious laboratory setting resembling the natural one. Petrocephalus catostoma, Cyphomyrus discorhynchus, and Hippopotamyrus sp. nov. differ characteristically in their behavior, such as in territorial defense, schooling, and joining members of other species during foraging. Comparing social encounters within and between species, the first evidence for interspecific electrocommunication among syntopic species was found. PMID- 10798202 TI - Mechanics and morphology of silk drawn from anesthetized spiders. AB - CO2 and N2 anesthetized Nephila spiders produced dragline silk with mechanical properties that differed from control silk as a function of time under anesthesia. Silk from CO2 spiders had a significantly lower breaking strain and breaking energy, significantly higher initial modulus, and marginally lower breaking stress. At the onset of anesthesia the silk diameter became highly variable. During deep anesthesia silk either became thinner or retained cross section but fibrillated. PMID- 10798203 TI - [Factors determining the modality selection in dialysis]. PMID- 10798204 TI - [Antihypertensive treatment in type II diabetes and diabetic nephropathy]. AB - Hypertension is very prevalent in patients with type II diabetes. Beside increasing the cardiovascular risk, hypertension has several deleterious effects on the kidney: hypertension promotes the development of diabetic nephropathy, the progression from microalbuminuria to overt diabetic nephropathy and progression to end stage renal disease. In this review, we analyze systematically the benefit of antihypertensive therapy in patients with type II diabetes, with either normo albuminuria, microalbuminuria or overt nephropathy. General considerations are developed about the general use of antihypertensive drugs in this population. An approach based on the prevention of the global or absolute cardiovascular risk is further recommended due to the very high cardiovascular burden in this diabetic patients. PMID- 10798205 TI - [Factors determining the choice of a modality of treatment by dialysis: a study of nine dialysis centers]. AB - BACKGROUND: The objective of this cross-sectional study in a population of 1472 dialysis patients was to identify the main factors involved in the choice of a specific option for dialysis therapy, taking into account three different types of criteria such as medical dependence (DM), nurse care requirement (SI) and independence for dialysis therapy (CA). METHODS: Each patient has been analysed, independently of present treatment modality, according to the above three criteria, namely DM, SI and CA. For each type of parameter, patients have been allocated to one of three levels, each level being established to evaluate whether dialytic treatment should be undertaken as hospital centre dialysis (HDC) or in a facility off the hospital. Level 3 of any one category corresponded to the inability of doing haemodialysis at home (HHD) or in self-care unit (AD). Level 2 included patients who could be treated in AD or by peritoneal dialysis (PD) with the assistance of a nurse. CAPD or HHD were considered as potential treatment modalities only in patients qualifying for level 1 of each criterion. RESULTS: In the patient population as a whole, the following treatment options were observed: HHD 3.6%, CAPD 6%, PD 1.8%, AD 16.3% and HDC 72.2%. For medical dependence (DM) there was a relatively even distribution for the three levels in six centres. In contrast, two centres were characterized by a predominance of DM level 3. Differences in DM levels between centres were greatly reduced when considering separately only those patients who were actually treated by CAPD, HDC and AD. SI levels were more uniformly distributed within all centres, and this was true for HCD and AD patients. When considering CA levels in HDC patients, a large predominance of CA level 3 was observed in all centres whereas CA level 1 was nearly in existent. CONCLUSION: The major finding of this study was that the inability or the refusal of dialysis patients to participate at treatment, independently of medical condition and nurse care requirement, was the main factor in the choice of hospital centre dialysis. PMID- 10798206 TI - [Renal and metabolic complications of lithium]. AB - Renal and metabolic adverse effects of lithium therapy are illustrated by the case report of a manic depressive woman aged 78 years, so treated for about 25 years. Long term lithium therapy with plasma lithium level in the therapeutic range impairs renal concentrating ability in 25-50% of the patients (when the total ingested amount reaches 100-200 mol, 700-1400 g). About 10-15% of the patients have overt nephrogenic diabetes insipidus (NDI) with elevated antidiuretic hormone plasma level and unresponsiveness to desmopressin. In rats, lithium treatment down regulates expression of the main water channel, aquaporin 2, in the renal collecting duct. NDI may be complicated by hypernatremic dehydration if the access to water is restricted, whatever the cause. Treatment of NID is best started with nonsteroidal antiinflammatory drugs, being then substituted for amiloride. Prolonged lithium therapy may induce chronic interstitial nephritis. In some patients this may result in mild or moderate non progressive chronic renal insufficiency. Acute lithium intoxication (with supratherapeutic doses) may be complicated by acute renal failure (ARF); even in the absence of ARF hemodialysis is indicated when plasma lithium level reaches 4 mmol/l or more. Other metabolic adverse effects of lithium therapy include: hypercalcemia due to hyperparathyroidism (in 5-10% of the patients); hypothyroidism (often latent); hyperthyroidism. In conclusion, these renal and metabolic adverse effects are generally mild or moderate, allowing the continuation of lithium therapy in most affected patients. PMID- 10798207 TI - [How to produce systematic reviews of excellent quality and to define evidence based nephrology]. AB - Since recent years, a number of systematic reviews have been published in the renal field. These publications will be used to develop the concept of evidence based nephrology. The design and quality of renal systematic reviews relies on technical skills and efficient trials search. The reasons for discrepancies that are sometimes reported between systematic reviews and controlled clinical trials are now better understood and may be partly explained by inadequate trial recovery and publication biases. The applicability of evidence findings is not always an easy task on patients level. The example of the low protein diet in chronic renal failure highlights such caveats. However, producing high level systematic reviews in the renal field may be of great help to clarify the use of controversial treatments and to design new studies in the renal field. PMID- 10798208 TI - [Guidelines for drug therapy in type II diabetes. French Agency for Health Safety in Health Products]. PMID- 10798209 TI - Diet-associated risks of disease and self-reported food consumption: how shall we treat partial nonresponse in a food frequency questionnaire? AB - Using data from a population-based case-control study on thyroid cancer, we studied two approaches of handling missing answers in a food frequency questionnaire when estimating food consumption and nutrient intakes. We analyzed the dietary reports of 165 cases and 248 control subjects. In the first approach, the omitted food items were considered as "null consumption." In the second approach, the missing answers were replaced with the median frequencies for subjects in the corresponding outcome category actually answering that specific food item. The results showed marginal differences between the two methods. In addition, the null consumption assumption was validated by means of complementary telephone interviews with a subsample of subjects who provided incomplete reports. Overall, the originally omitted answers actually corresponded to very rare consumption for 54.2% of cancer patients and for 54.7% of controls. However, this "true" proportion of null consumption varied greatly between different food items (range 0-96%) and between food groups (range 14-82%). To interpret the omitted self-reports of food consumption as indication of "zero consumption" is quite reasonable when the investigation does not focus on dietary items widely consumed in the source population. PMID- 10798210 TI - Waist-to-hip ratio, weight gain, and dietary and serum selenium are associated with DNA content flow cytometry in Barrett's esophagus. AB - This cross-sectional study reports associations between anthropometric measures, serum antioxidant concentrations, and present diet with measures of elevated cell proliferation in 51 patients with Barrett's esophagus. Cell proliferation was assessed as fractions of cells in the S and G2 phases, measured in biopsies of Barrett's tissue and analyzed by DNA content flow cytometry. Elevated proportions in the S and G2 phases predict progression to adenocarcinoma. The percentage of cells in the S phase was positively associated with waist-to-hip ratio (r = 0.33, p < 0.05) and negatively associated with serum and dietary selenium (r = -0.34 and -0.32, respectively, p < 0.05). The percentage of cells in the G2 phase was positively associated with weight change from age 25 (r = 0.39, p < 0.01) and negatively associated with dietary selenium (r = -0.31, p < 0.05). Selenium from breads and grains was negatively associated with the percentage of cells in the S phase (r = -0.41, p < 0.01) and the percentage of cells in the G2 phase (r = 0.41, p < 0.01). These results suggest that increasing weight gain in adulthood, increasing waist-to-hip ratio, and decreasing dietary selenium intake and serum levels increase the risk of progression of Barrett's esophagus to adenocarcinoma. PMID- 10798211 TI - Inverse association of soy product intake with serum androgen and estrogen concentrations in Japanese men. AB - The cross-sectional relationships of soy product intake and serum testosterone, estrone, estradiol, sex hormone-binding globulin, and dihydrotestosterone were examined in 69 Japanese men. Soy product intake was estimated from a semiquantitative food frequency questionnaire. Serum estradiol concentration was significantly inversely correlated with soy product intake (r = -0.32, p = 0.009), and serum estrone concentration was nonsignificantly inversely correlated with soy product intake (r = -0.24, p = 0.05) after controlling for age, body mass index, smoking status, and ethanol intake. Total and free testosterone concentrations were inversely correlated with soy product intake after controlling for the covariates, but these correlations were of border line significance (r = -0.25, p = 0.05 and r = -0.25, p = 0.06, respectively). Similar correlations were observed for these hormones with isoflavone intake from soy products. The data suggest that soy product intake may be associated with the endogenous hormone levels in Japanese men. PMID- 10798212 TI - Selenoprotein P in plasma in relation to cancer morbidity in middle-aged Swedish men. AB - The premorbid level of selenoprotein P in plasma from subjects with cancer at different sites was compared with that from control subjects in a nested case control study. A health screening of 12,500 middle-aged men was performed during 1974-1982 in Malmo, Sweden, and from the 400 cancer cases that were identified during follow-up until the end of 1988, 302 plasma samples were available for analysis of selenoprotein P. Two living controls per case of the same screening day and age were chosen. Selenoprotein P levels in subgroups of major cancer sites were lower in cases than in controls for the respiratory tract (1.20 and 1.30 arbitrary units, respectively; p < 0.05) cancer group. The odds ratio for overall cancer risk in the lowest quintile of selenoprotein P level compared with that in the highest was 5.2 [p (for trend) = 0.01]. In subgroups of major cancer sites, the odds ratios for cancer risk in the lowest tertile compared with the highest were 6.0 [p (for trend) = 0.004] in the respiratory tract and 3.4 [p (for trend) = 0.002] in the digestive tract. In cases + controls, selenoprotein P was lower in smokers than in nonsmokers (p < 0.05). Selenoprotein P was significantly correlated to plasma albumin, fasting blood glucose, and body mass index and inversely correlated to plasma alpha 1-antitrypsin and gamma-glutamyl transferase. The results suggest that a low plasma selenoprotein P level is associated with higher future risk of respiratory and digestive tract cancer in middle-aged men. PMID- 10798213 TI - Interindividual variation in metabolism of soy isoflavones and lignans: influence of habitual diet on equol production by the gut microflora. AB - The soy isoflavones, daidzein and genistein, and the lignans, matairesinol and secoisolariciresinol, are phytoestrogens metabolized extensively by the intestinal microflora. Considerable important evidence is already available that shows extensive interindividual variation in isoflavone metabolism, and we have investigated the extent of this variation in a crossover study of a soy containing food low or high in isoflavones (each treatment period lasted for 17 days, and the 2 treatment periods were separated by a 25-day washout period) in 24 healthy subjects [19 women and 5 men, mean age 30 yr, range 19-40, mean body mass index 22.5 +/- 3.5 (SD) kg/m2]. There was a 16-fold variation in total isoflavonoid excretion in urine after the high-isoflavone treatment period. The variation in urinary equol excretion was greatest (664-fold), and subjects fell into two groups: poor equol excretors and good equol excretors (36%). A significant negative correlation was found between the proportion of energy from fat in the habitual diet and urinary equol excretion (r = -0.55; p = 0.012). Good equol excretors consumed less fat as percentage of energy than poor excretors (26 +/- 2.3% compared with 35 +/- 1.6%, p < 0.01) and more carbohydrate as percentage of energy than poor excretors (55 +/- 2.9% compared with 47 +/- 1.7%, p < 0.05). Interindividual variation in the urinary excretion of O-desmethyl-angolensin (O DMA) was also apparent (76-fold after the high-isoflavone treatment period), but there was no relationship between equol excretion and O-DMA excretion. Enterolactone was the major lignan metabolite in urine and plasma but showed less interindividual variation than equol and O-DMA. It is suggested that the dietary fat intake decreases the capacity of gut microbial flora to synthesize equol. PMID- 10798214 TI - Effect of an alpha-linolenic acid-rich diet on rat mammary tumor growth depends on the dietary oxidative status. AB - To investigate whether the oxidative status of an 18:3(n-3) polyunsaturated fatty acid (PUFA)-enriched diet could modulate the growth of chemically induced rat mammary tumors, three independent experiments were performed. Experiments I and II examined the variation of tumor growth by addition of antioxidant (vitamin E) or a prooxidant system (sodium ascorbate/2-methyl-1,4-naphthoquinone) to a 15% linseed oil diet rich in 18:3(n-3). Experiment III addressed the role of PUFA in the tumor growth modulation by vitamin E. For this purpose, we compared the effect of vitamin E in 15% fat diets containing a high level of 18:3(n-3) (linseed oil, high-PUFA diet) or devoid of 18:3(n-3) (hydrogenated palm/sunflower oil, low-PUFA diet). In Experiments I-III, tumor growth increased in the presence of vitamin E compared with control (without vitamin E). Furthermore, it decreased when prooxidant was added. In contrast, no difference was observed when the diet was low in PUFA, suggesting that sensitivity of PUFA to peroxidation may interfere with tumor growth. This observation was supported by growth kinetic parameter analysis, which indicated that tumor growth resulted from variations in cell loss but not from changes in cell proliferation. These data show that, in vivo, PUFA effects on tumor growth are highly dependent on diet oxidative status. PMID- 10798215 TI - Effects of bleomycin on liver antioxidant enzymes and the electron transport system from ad libitum-fed and dietary-restricted female and male Fischer 344 rats. AB - Dietary restriction (DR) is the only known intervention that delays aging and age related diseases. Mechanisms proposed to explain this DR effect include a decline in free radical production and an increase in free radical detoxification. In the present study the effect of bleomycin (BLM) as a reactive oxygen species generating antitumor drug has been evaluated on antioxidant enzymes and the electron transport system in different cellular fractions of liver in female and male Fischer 344 rats. Animals were fed ad libitum (AL) or 60% of the AL intake (DR) and were given a single intraperitoneal injection of 2.5, 5, or 10 mg BLM/kg body wt. After four weeks, BLM significantly increased glutathione peroxidase and lactate dehydrogenase activities in liver cytosol of female AL rats and increased activity even more in male rats. Similar changes were also noted for glutathione reductase and glucose 6-phosphate dehydrogenase activities in BLM-treated AL rats. In liver mitochondria, glutathione peroxidase was increased in female and male AL rats but was increased more in female rats. Drug treatment had no significant effect on these enzyme activities in cytosolic or mitochondrial fractions of DR animals. Profound effects of BLM were noted in activities of complexes I, III, and IV of the electron transport system in AL and DR female and male rats; however, complex II demonstrated no significant diet or treatment effect. Induced antioxidant enzyme activities in BLM-treated AL rats may be a response to excessive free radical generation due to BLM metabolism in AL animals that is mitigated by DR. Furthermore, dysfunction of the electron transport system might suggest its role in a secondary generation of free radicals during BLM metabolism contributing to its toxicity. PMID- 10798216 TI - Tea as a potential chemopreventive agent in PhIP carcinogenesis: effects of green tea and black tea on PhIP-DNA adduct formation in female F-344 rats. AB - The heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is formed during the cooking of proteinaceous animal foods (meat, chicken, and fish). PhIP is a carcinogen in the Fischer 344 (F-344) rat; it induces mammary tumors in female rats and lymphomas and colon and prostate tumors in male rats. In F-344 rats, PhIP forms DNA adducts in various organs, including the target organs. Inhibition of PhIP-DNA adduct formation is likely to lead to inhibition of PhIP tumorigenicity. We have examined the chemopreventive properties of green tea and black tea in PhIP carcinogenesis by evaluating their effects on PhIP-DNA adduct formation in the female F-344 rat. Young adult animals were maintained on powdered AIN-76A diet while receiving regular drinking water or 2% (wt/vol) infusions of green tea or black tea for a total of six weeks. During Weeks 3, 4, and 5, all animals received PhIP by gavage (1 mg/kg/day). Three rats per group were euthanized on Days 1 and 8 after termination of PhIP exposure. DNA was isolated from a number of organs and analyzed for PhIP-DNA adducts by 32P postlabeling assays. Compared with animals on regular drinking water, PhIP-DNA adduct formation was inhibited in small intestine, colon, liver, and mammary epithelial cells (MECs) of animals receiving green tea or black tea as the sole source of drinking fluid. Green tea inhibited adduct formation in colon, liver, and MECs (33.3-80.0%) on both days, but only on Day 8 (54.4%) in small intestine. Black tea inhibited adduct formation on both days in liver (71.4-80.0%), on Day 1 in colon (40.0%), and on Day 8 in small intestine (81.8%); it had no effect on MEC adducts. Neither green tea nor black tea had an effect on adduct levels in pancreas, lungs, white blood cells, heart, kidneys, spleen, cecum, or stomach. Similarly, these teas did not affect the rate of adduct removal (percent change from Day 1 to Day 8) in any organ. It is concluded that green tea and black tea are potential chemopreventive agents in PhIP-induced tumorigenesis in the F-344 rat. PMID- 10798217 TI - Antitumor activity of astaxanthin and its mode of action. AB - Astaxanthin, a carotenoid without vitamin A activity, may exert antitumor activity through the enhancement of immune responses. Here, we determined the effects of dietary astaxanthin on tumor growth and tumor immunity against transplantable methylcholanthrene-induced fibrosarcoma (Meth-A tumor) cells. These tumor cells express a tumor antigen that induces T cell-mediated immune responses in syngenic mice. BALB/c mice were fed astaxanthin (0.02%, 40 micrograms/kg body wt/day in a beadlet form) mixed in a chemically defined diet starting zero, one, and three weeks before subcutaneous inoculation with tumor cells (3 x 10(5) cells, 2 times the minimal tumorigenic dose). Three weeks after inoculation, tumor size and weight were determined. We also determined cytotoxic T lymphocyte (CTL) activity and interferon-gamma (IFN-gamma) production by tumor draining lymph node (TDLN) and spleen cells by restimulating cells with Meth-A tumor cells in culture. The astaxanthin-fed mice had significantly lower tumor size and weight than controls when supplementation was started one and three weeks before tumor inoculation. This antitumor activity was paralleled with higher CTL activity and IFN-gamma production by TDLN and spleen cells in the astaxanthin-fed mice. CTL activity by TDLN cells was highest in mice fed astaxanthin for three weeks before inoculation. When the astaxanthin-supplemented diet was started at the same time as tumor inoculation, none of these parameters were altered by dietary astaxanthin, except IFN-gamma production by spleen cells. Total serum astaxanthin concentrations were approximately 1.2 mumol/l when mice were fed astaxanthin (0.02%) for four weeks and appeared to increase in correlation with the length of astaxanthin supplementation. Our results indicate that dietary astaxanthin suppressed Meth-A tumor cell growth and stimulated immunity against Meth-A tumor antigen. PMID- 10798218 TI - Inhibition by ginseng of 1,2-dimethylhydrazine induction of aberrant crypt foci in the rat colon. AB - The modifying effects of dietary administration of ginseng on the induction and development of 1,2-dimethylhydrazine (DMH)-induced aberrant crypt foci (ACF) were investigated in Fischer 344 (F-344) rats. In Experiment 1, starting at six weeks of age, 65 rats were injected with DMH or saline alone once a week for four weeks. Rats in Groups 1 and 2 were fed diets containing 1% ginseng for five weeks, starting one week before the first dose of DMH. Animals in Groups 3 and 4 received ginseng for eight weeks after DMH treatment; Group 5 served as a carcinogen control group. In Experiment 2, 60 rats were injected with DMH or saline alone four times at one-week intervals. They were also fed diets containing 1% ginseng or the control diet throughout the 30 days of the experiment. In Experiment 1, numbers of foci with at least four crypts were significantly reduced in Group 2 treated with red ginseng during the initiation phase (p < 0.005). In Experiment 2, treatment with red ginseng also resulted in a decrease in the total number of DMH-induced ACF accompanied by a reduction in 5 bromo-2'-deoxyuridine labeling indexes in colonic crypts comprising ACF (p < 0.005 and p < 0.05, respectively). These findings suggest that dietary administration of red ginseng in combination with DMH suppresses colon carcinogenesis of rats, and the inhibition may be associated, in part, with suppression of cell proliferation in the colonic mucosa. PMID- 10798219 TI - Effect of beta-sitosterol, a plant sterol, on growth, protein phosphatase 2A, and phospholipase D in LNCaP cells. AB - Previous work from this laboratory suggests an activation of the sphingomyelin cycle as a mechanism for growth inhibition with the incorporation of beta sitosterol (SIT) into human prostate cancer LNCaP cells. In the present study we examined two key enzymes that have been shown to play a role in the sphingomyelin cycle. Dietary sterols (SIT and cholesterol) were compared for their effect on LNCaP cell growth, phospholipase D (PLD) activity, and protein phosphatase 2A (PP 2A) activity and expression. PP 2A has been suggested as a direct in vitro target of ceramide action on cell growth and apoptosis. Ceramide also inhibits phorbol myristate acetate-stimulated PLD. SIT (16 microM) increased PP 2A activity by 50% compared with cholesterol treatment in LNCaP prostate cells; however, SIT did not alter protein levels of PP 2A. There was an increase in PLD activity in the presence of phorbol myristate acetate in cells supplemented with 16 microM SIT compared with those supplemented with cholesterol after five days of treatment. The present study suggests that the activation of PP 2A added support to the role of the activation of the sphingomyelin cycle by SIT treatment. However, the increase in PLD activity, which was modest but significant, with SIT supplementation suggests that this pathway may be modulated by other mechanisms. This includes the incorporation of SIT into cell membranes that may alter fluidity and, thus, influence the activation of membrane-bound enzymes such as PLD. PMID- 10798220 TI - Effect of colonic luminal components on induction of apoptosis in human colonic cell lines. AB - Apoptosis is central to cell number regulation in the colonic epithelium, and interest in its role in colon carcinogenesis has been growing rapidly. It thus becomes of interest to characterize luminal components, possibly of dietary origin, that may influence this process. We have investigated the sensitivity of two human colonic cell lines, the human adenocarcinoma cell line (HT-29) and the human fetal colonic mucosa cell line (FHC), to induction of apoptosis by sodium butyrate, bile acids, and human fecal water fractions. The apoptotic effect has been studied by 1) morphological changes in cells examined by fluorescence microscopy, 2) DNA fragmentation analysis by gel electrophoresis, 3) flow cytometry analysis of DNA strand breaks assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL), and 4) poly(ADP ribose) polymerase cleavage by Western blot. Sodium butyrate and bile acids induced a time- and concentration-dependent apoptosis in both cell lines. Quantitation of this effect, by use of the TUNEL assay, indicated that deoxycholic acid was most effective in inducing this effect at lower concentrations and at shorter times. Apoptotic effects were also observed, in both cell lines, when the cells were exposed to intact human fecal waters (the fecal fraction in direct contact with the epithelium) and their lipid extracts, with the intact samples being more effective. Although all fecal waters examined induced apoptosis, quantitation of the effect by the TUNEL assay indicated that the ability to induce apoptosis differed markedly between samples. Induction of apoptosis by the fecal waters was not correlated to cytotoxicity but was negatively correlated to the pH of the samples. Interestingly, the cells derived from the fetal mucosa (FHC) were consistently less sensitive to apoptotic effects of the luminal components than the tumor-derived cells (HT-29). Thus human fecal water fractions induce apoptosis in colonic cells, and this effect is not due to lipid components alone. PMID- 10798221 TI - Vitamin E succinate induces apoptosis in human prostate cancer cells: role for Fas in vitamin E succinate-triggered apoptosis. AB - The apoptosis-triggering properties of vitamin E succinate (VES, RRR-alpha tocopheryl succinate) for human LNCaP and PC-3 prostate carcinoma cells and normal PrEC human prostate epithelial cells were investigated. LNCaP and PC-3 cells were sensitive to VES-induced apoptosis, with 100% and 60% of cells undergoing apoptosis after three days of treatment with 10 micrograms of VES/ml, respectively. PrEC cells were resistant to VES-induced apoptosis. Treatment of prostate cells with agonistic anti-Fas antibody triggered apoptosis in approximately 50% of PC-3 cells within 48 hours, whereas LNCaP and PrEC cells were resistant. Prostate cells simultaneously treated with VES and agonistic anti Fas antibodies revealed 1) no effect on PrEC cells, 2) an additive effect on Fas sensitive PC-3 cells, and 3) a synergistic effect on LNCaP cells. VES treatment of LNCaP cells caused depletion of cytosolic 43-kDa Fas, enhanced membrane levels of 43-kDa Fas, and induced Fas sensitivity. PC-3 cells expressed high levels of membrane 43-kDa Fas that were enhanced by VES treatments. Fas ligand expression by LNCaP cells was enhanced by VES treatments. In summary, VES triggers apoptosis in human prostate carcinoma cells but not normal prostate cells in vitro, and VES modulates Fas signaling. PMID- 10798222 TI - Lycopene interferes with cell cycle progression and insulin-like growth factor I signaling in mammary cancer cells. AB - Recent studies have shown that high insulin-like growth factor I (IGF-I) blood level is a risk factor in breast and prostate cancer. The aim of this study was to determine whether the mitogenic activity of IGF-I in mammary cancer cells can be reduced by the dietary carotenoid lycopene. The anticancer activity of lycopene, the major tomato carotenoid, has been suggested by in vitro, in vivo, and epidemiological studies. Growth stimulation of MCF7 mammary cancer cells by IGF-I was markedly reduced by physiological concentrations of lycopene. The inhibitory effects of lycopene on MCF7 cell growth were not accompanied by apoptotic or necrotic cell death, as determined by annexin V binding to plasma membrane and propidium iodide staining of nuclei in unfixed cells. Lycopene treatment markedly reduced the IGF-I stimulation of tyrosine phosphorylation of insulin receptor substrate 1 and binding capacity of the AP-1 transcription complex. These effects were not associated with changes in the number or affinity of IGF-I receptors, but with an increase in membrane-associated IGF-binding proteins, which were previously shown in different cancer cells to negatively regulate IGF-I receptor activation. The inhibitory effect of lycopene on IGF signaling was associated with suppression of IGF-stimulated cell cycle progression of serum-starved, synchronized cells. Moreover, in cells synchronized by mimosine treatment, lycopene delayed cell cycle progression after release from the mimosine block. Collectively, the above data suggest that the inhibitory effects of lycopene on MCF7 cell growth are not due to the toxicity of the carotenoid but, rather, to interference in IGF-I receptor signaling and cell cycle progression. PMID- 10798223 TI - N-methoxyindole-3-carbinol is a more efficient inducer of cytochrome P-450 1A1 in cultured cells than indol-3-carbinol. AB - The well-documented reduction of cancer risk by high dietary cruciferous vegetable intake may in part be caused by modulation of cytochrome P-450 (CYP) expression and activity by indoles. The purpose of the present experiments was to study the mechanism of CYP 1A1 induction by N-methoxyindole-3-carbinol (NI3C) in cultured cells and to compare the CYP-inducing potential of NI3C and indole-3 carbinol (I3C) administered to rats. NI3C induced 7-ethoxyresorufin-O-deethylase (EROD) activity in Hepa-1c1c7 cells in a concentration-dependent manner with 10 fold higher efficiency than I3C. Inasmuch as NI3C induced binding of the aryl hydrocarbon receptor (AhR) to the dioxin-responsive element and induced expression of endogenous CYP 1A1 mRNA and an AhR-responsive chloramphenicol acetyl transferase construct, we conclude that NI3C can activate the AhR. Besides the induction of CYP 1A1, we observed an inhibition of EROD activity, with a concentration causing 50% inhibition of 6 microM. Oral administration of NI3C at 570 mumol/kg body wt to male Wistar rats increased the hepatic CYP 1A1 and 1A2 protein levels, as well as the EROD and 7-methoxyresorufin O-demethylase activities at 8 and 24 hours after administration, but the responses were less pronounced than after administration of I3C at 570 mumol/kg body wt. Furthermore, NI3C did not induce hepatic 7-pentoxyresorufin O-depentylase activity, as I3C did. Ascorbigen, another indolylic compound formed during degradation of glucobrassicin in the presence of ascorbic acid, was tested in the same animal model, and ascorbigen only weakly induced hepatic CYP 1A1 and 1A2, but not CYP 2B1/2. In conclusion, NI3C is a more efficient inducer of CYP 1A1 in cultured cells than I3C but is less active when administered to rodents. PMID- 10798225 TI - [Tendon injuries: on the threshold of a new era]. PMID- 10798224 TI - Lambda-carrageenan-induced inhibition of gap-junctional intercellular communication in rat liver epithelial cells. AB - lambda-Carrageenan, a food additive extracted from red seaweed, is widely used as an emulsifier, stabilizer, or thickener. Previously, it has been shown that carrageenan could play a role in carcinogenesis. However, the mechanism by which it might influence the multimechanism, multistep process of carcinogenesis is not known. Gap-junctional intercellular communication (GJIC) has been associated with maintaining homeostatic regulation of cell proliferation and differentiation. Most cancer cells have dysfunctional GJIC, and many tumor-promoting chemicals, growth factors, and oncogenes can downregulate GJIC. The experiments in this study were designed to test the hypothesis that carrageenan might function as a tumor-promoting chemical by inhibiting GJIC. To test this hypothesis, nontumorigenic rat liver epithelial cells were exposed to carrageenan, and GJIC was measured. Western blot analysis and immunofluorescent staining were used to monitor the phosphorylation and localization of connexin 43. The data revealed inhibition of GJIC by carrageenan similar to that by the well-documented tumor promoter phorbol ester. However, the phosphorylation and localization of connexin 43 were not altered. Although the mechanism by which carrageenan inhibits GJIC is unknown, carrageenan's influence on the carcinogenic process might be via its ability to be a tumor promoter. PMID- 10798226 TI - [Pathology of acute and chronic tendon injuries]. AB - The load limit of a tendon is expressed by the not linear force/lengthening behavior. It is determined by the interrelation between load and load-bearing capacity. The load of a tendon results from exogenous, endogenous and temporal factors. The property factors and the aging influence the load-bearing capacity of a tendon. Partial ruptures show necrosis, calcification and fiber swelling. The attachment area shows a mineralization of the cartilage zone, a loss of the collagen fibrils and calcifications. The tendon rupture knows extrinsic and intrinsic causes. Primary are micro and macrovascular tendon alterations, usually favour the rupture of tendons. PMID- 10798227 TI - [Current aspects of tendon healing]. AB - This review describes structure, function and healing of tendinous tissue and discusses new biologically based treatment options to modulate tendon healing. The repair process after tendon rupture results in a morphologically different and biomechanically inferior structure compared to a normal tendon. The collagen fibril diameters are decreased months after the traumatic lesion and show also different phenotypes. We know that cytokines and growth factors are key components for normal tissue development and regulate wound healing processes. Some growth factors have been detected to influence tenocytes by promoting cell proliferation and matrix synthesis. Application of the adequate growth factors at certain periods during the repair process might improve the healing result after tendon rupture. However, most of these growth factors are proteins which are rapidly metabolized by the organism. Transfer of growth factor genes into tenocytes might eliminate this problem by a continuous local release of growth factors at the healing site. PMID- 10798228 TI - [Tendon suture: surgical techniques]. AB - Tendon sutures belong to the bases of surgical activity. Particularly in the last decades these sutures experienced a substantial upswing by the introduction of new techniques and materials. Target is to reestablish the tendon function. In order to achieve this target, it requires knowledge of tendon healing. An outline of suture materials, in particular specific suture techniques as well as the subsequent treatment is given. With different localizations of the tendon lesions one deals. PMID- 10798229 TI - [Rotator cuff tears: diagnosis and therapy]. AB - Since the life span of our society is increasing and the expectation of high functional demands is growing more and more older people take part in sports activities emphasises the necessity of early diagnosis of rotator cuff lesions and their correct treatment. The goal of any diagnostic means should be a proper treatment of rotator cuff lesions according to the detected pathology. New arthroscopic techniques to surgically reconstruct rotator cuff tears have brought the operative repair techniques another step further. In cases of large and complex cuff tears the surgeon can individually select an adequate reconstruction procedure which includes anatomical-, partial-, tendon-transfer and muscle transfer repairs. In cases of irreparable tears with cuff-tear arthropathy and consecutive superior migration of the humeral head hemiarthroplasty or the inverse deltoid prosthesis have to be considered. Based on clinical, radiological and arthroscopic findings we developed algorithms that will lead to the best suited procedure for a given lesion. PMID- 10798230 TI - [Biceps tendon: diagnosis, therapy and results after proximal and distal rupture]. AB - Ruptures of the long head of the M. biceps humeri are commonly caused by degenerative changes within the tendon. They are associated with pathologies of the subacromial space. The loss of power regarding elbow flexion and supination amounts to 8 to 21% after conservative treatment. Refixation offers a small but evident improvement of flexion and supination power. Especially endurance is improved. The number of cases with remaining light or marked weakness is reduced by more than 50%. Deformity by the slipped muscle can be corrected effectively. Function of the glenohumeral joint can only be improved if associated subacromial problems are identified and treated simultaneously. As complications are uncommon surgery should be recommended to young and active patients and should at least be offered to less active patients. Ruptures of the distal tendon are less common. Thirteen patients were re-examined after operative repair for distal biceps tendon avulsion and 277 reported cases were reviewed. After conservative management (n = 20) the power of flexion will remain reduced by 30%-40%, that of supination by more than 50%. The loss of flexion power, as well as the deformity can be diminished by attachment of the distal biceps to the brachialis muscle (n = 22). The anatomic re-insertion (n = 248) additionally reduces the loss of supination power to 0%-25%, but bears a higher risk of complications. Using the 'double-incision technique' (n = 105 of 248) does not decrease the risk of naval lesions but increases the incidence of radioulnar synostosis. The use of suture anchors provides a nice way of fixation of the tendon but does not facilitate the approach to the tuberosity. The distal biceps tendon rupture should be treated operatively. The adequate method of repair is to be determined individually. PMID- 10798232 TI - [Quadriceps and patellar tendon ruptures]. AB - Ruptures of the quadriceps as well as the patellar tendon occur in low frequency, but cause major functional deficits of the leg. These injuries usually require operative treatment. Acute quadriceps tendon ruptures are treated by suture repair, using heavy sutures guided through bone tunnels in the patella. Chronic defects and neglected cases require a local tendon transfer, either by a quadriceps tendon turn-down or by a V-Y-plasty of the quadriceps tendon. Ruptures of the patellar tendon are treated by suture of the tendon stumps plus an reinforcement procedure protecting the tendon and avoiding secondary patella alta. Patello-tibial fixation may be achieved by a cerclage technique using wire or an autologous tendon strip, alternatively a patello-tibial external fixator can be applied. In chronic and neglected cases, patellar tendon reconstruction is performed with autologous tendon grafts or with soft tissue allografts. The graft must be protected by a patello-tibial fixation for the first weeks. PMID- 10798231 TI - [Flexor and extensor tendon injuries of the hand]. AB - Injuries of the flexor tendon are usually open injuries. In most cases primary treatment is therefore seldom performed by orthopaedic surgeons. This is different for injuries of the extensor tendon. Three quarters of injuries of the extensor tendon are closed injuries. Therefore we do go into primary and secondary reconstruction of the flexor tendon, but focus on primary and secondary reconstruction of the extensor tendon in the most common zones (1, Th I, 3, Th III). PMID- 10798233 TI - [Achilles tendon rupture]. AB - The treatment of acute of Achilles tendon rupture experienced a dynamic development in the last ten years. Decisive for this development was the application of MRI and above all the ultrasonography in the diagnostics of the pathological changes and injuries of tendons. The question of rupture morphology as well as different courses of healing could be now evaluated objectively. These advances led consequently to new modalities in treatment concepts and rehabilitation protocols. The decisive input for improvements of the outcome results and particularly the shortening of the rehabilitation period came with introduction of the early functional treatment in contrast to immobilizing plaster treatment. In a prospective randomized study (1987-1989) at the Trauma Dept. of the Hannover Medical School could show no statistical differences comparing functional non-operative with functional operative therapy with a special therapy boot (Variostabil/Adidas). The crucial criteria for therapy selection results from the sonographically measured position of the tendon stumps in plantar flexion (20 degrees). With complete adaptation of the tendons' ends surgical treatment does not achieve better results than non-operative functional treatment in term of tendon healing and functional outcome. Regarding the current therapeutic standards each method has is advantages and disadvantages. Both, the operative and non-operative functional treatment enable a stable tendon healing with a low risk of re-rupture (1-2%). Meanwhile there is consensus for early functional after-treatment of the operated Achilles' tendons. There seems to be a trend towards non-operative functional treatment in cases of adequate sonographical findings, or to minimal invasive surgical techniques. PMID- 10798234 TI - [Injuries of the tibialis posterior tendon: diagnosis and therapy]. AB - Complete traumatic rupture of the tibialis posterior tendon is absolutely rare. From the scarce case reports in the medical literature a extreme pronation abduction or pronation-external rotation mechanism according to the Lauge-Hansen classification can be presumed, leading to a malleolar fracture because of forced pronation, external rotation and dorsiflexion of the foot. With primary suture the prognosis is favorable. Traumatic dislocations, mostly with luxatio pedis sub talo, are treated by atraumatic reduction and refixation of the retinaculum. Again, the prognosis is favorable. Incomplete traumatic rupture of the tibialis posterior tendon with development of posttraumatic pes plano valgus, according to case reports and our own experience result from severe pronation-external rotation-soft tissue injuries as well as with pronation-abduction or pronation external rotation-type ankle fractures. In these cases no macroscopic rupture of the tendon is evident, however occult interstitial micro-ruptures can occur because of excessive stretching, which can be determined histologically. If conservative measures fail, a modified Evans osteotomy to lengthen the lateral foot column is indicated. Degenerative complete and incomplete ruptures of the tibialis posterior tendon are predominantly seen in women more than 42 years old. Staging of this entity can be achieved with clinical tests (muscular force, external rotation), ultrasound, weight-bearing x-rays, CT and MRT. According to the degree of decompensation of tendon function, treatment consists of augmentation, modified Evans procedure or triple arthrodesis of the hind-foot. PMID- 10798235 TI - [Effects of the cementing technique on cementing results concerning the coxal end of the femur]. AB - The purpose of this study was to determine the influence of jet-lavage and cement pressurising techniques upon cement penetration into proximal femoral cancellous bone. In a cadaver study 60 left human cadaver femora were used for implantation of cemented stem components. Four different groups of cementing techniques were generated, the allocation to the groups was randomized. Bone lavage was carried out either using jet-lavage or manual syringe lavage, cement application differed with regard to the amount of pressurisation used. Five different stem designs were used. Radiographs were taken and horizontal sections were obtained at predefined levels (2 cm) using a diamond saw. Microradiographs were taken and analysed using image analysis to assess cement penetration into cancellous bone. In an additional study the influence of jet-lavage (1000 ml) versus syringe lavage (1000 ml) was studied in 11 paired human cadaver femora. The specimens were imbedded in specially designed pots. Bone cement was applied in a retrograde manner and subjected to a standard pressure protocol with a constant force of 3000 N. The analysis protocol was identical to the main experiment. Both jet lavage and pressurisation of bone cement significantly improved the penetration of cement into cancellous bone (p = 0.027 and p = 0.003, respectively). In the presence of strong, dense cancellous bone the findings were more pronounced. In the additional comparative study cement intrusion was significantly better (p < 0.001) in the jet-lavage group. We did not observe an influence of the stem type upon outcome (penetration). The use of jet-lavage yields significantly improved cement penetration and should be regarded mandatory in cemented total hip arthroplasty. High pressurising techniques are effective means to improve the interdigitation between cancellous bone and cement, but should only be administered in combination with jet-lavage to reduce the risk of thrombo-embolic complications. PMID- 10798236 TI - [Is homeopathic medicine equivalent to diclofenac?! An opinion statement against misleading]. PMID- 10798237 TI - Glycosidase inhibitors and their chemotherapeutic value, Part 1. AB - The various compounds that have been investigated as glycosidase inhibitors are reviewed. The first of three parts of this review article covers the following classes of compounds: sugar, lactones and hydroximolactones, glycosyl halides, oligosaccharides, glycosides and their derivatives, deoxy thiosugar derivatives, thioglycosides, deoxy amino and guanidino sugars, glycosylamines, anhydrosugars and their analogues, deoxysugars, glycals, C-glycosides and C-nucleosides. PMID- 10798238 TI - Nitrogen bridgehead compounds, facile synthesis of bioactive cyanopyrimido[1,2 a]pyrimidinones. AB - Synthesis of some new cyanopyrimido[1,2-a]pyrimidinones 5-22 have been achieved via interaction of 2-amino-6-anisyl-5-cyano-4(3H)-pyrimidinone (1) with some heterocycles having a vicinal chloroester, chlorocyano or mercaptocyano group, dimethyl acetylenedicarboxylate, active methylene compounds, ethyl 2-acetyl-3 anisylpropenoate, ethyl 3-aryl-3-cyanopropenoates, ethyl 2-cyano-3-ethoxyacrylate and some enones or enals. Some of the isolated products were subjected to biological screening tests. PMID- 10798239 TI - Synthetic approaches towards benzo[h]quinoline-3-carbonitriles. AB - 2-Alkoxybenzo[h]quinoline-3-carbonitriles 2 were readily synthesized via Dimroth rearrangement of 2-amino-4-aryl-5,6-dihydro-4H-naphtho[1,2-b]pyran-3 carbonitriles 5 induced by ethanolic and methanolic KOH. Compounds 2 were also obtained by the reaction of 2-arylmethylidene-3,4-dihydro-1(2H)-naphthalenones 1 with malononitrile or of ylidenemalononitriles 4 with 1,2,3,4-tetrahydro-1 naphthalenone in ethanolic and methanolic KOH. The molluscicidal activity of the products towards Biomphalaria alexandrina snails was screened. PMID- 10798240 TI - [Reactions of 4,5-dihydro-4-oxo-1H-pyrido(3,2-b)indol-2-carboxylic acid ester]. AB - The title compounds 5 were saponificated and decarboxylated, N- and O-alkylated; the ester group was reduced to the primary alcohol and than oxidized to the aldehyde. In the same manner the N- and O-methylated products 9 and 10 were derivatized. The tetrazoles 24 and 25 were synthesized from the aldehydes 18 and 19 via the aldoximes and nitriles. The aldehydes 18 and 19 reacted with beta amino crotonic acid esters in the Hantzsch pyridine synthesis to mixtures, from which the 1,4-dihydropyridines (DHP) 26 and 27 as well as the ylidenes from methyl acetoacetate 31 and 32 and in the case of 18 the oxidation product 30 of a 1,2-DHP could be isolated. The pyridines 28 and 29 were obtained by chemical dehydrogenation of the 1,4-DHP 26 and 27. By anodic oxidation of 26 and 27 by means of difference pulse voltammetry not only an one step two-electronic dehydrogenation yielding the corresponding pyridines took place, but also a two step one-electronic oxidation of the vinylogous hydrazine structure to diiminium salts occurred. PMID- 10798242 TI - Development and validation of an indirect enzyme-linked immunosorbent assay (ELISA) for the nonsteroidal anti-inflammatory drug S-ibuprofen. AB - An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the nonsteroidal anti-inflammatory drug (NSAID) S-ibuprofen. Conjugates for immunization were prepared by linking S-ibuprofen via the spacer 4-aminobutyric acid to bovine serum albumin as well as to a novel synthetic lipopeptide using the N-hydroxysuccinimide/dicyclohexylcarbodiimide method. Immunization with these immunogens was carried out in New Zealand rabbits. A poly-L-lysine-S-ibuprofen conjugate was used as a hapten-carrier for coating the surface of the microtiter plates with the hapten. Horse-radish peroxidase labeled anti-rabbit IgG served as secondary antibody using hydrogen peroxide and ABTS as substrates. The characterization of the polyclonal antiserum with compounds of analogous structure demonstrated that the antiserum possesses a very high specificity for S ibuprofen (cross-reactivity < 0.14-1.4%). Additional cross-reactivity experiments using R-ibuprofen (cross-reactivity 50.5%), ibufenac (58%) and isopropylphenylacetic acid (6.4%) were carried out to obtain more detailed information about the antigenic recognition concerning the chiral center. The results indicated that the polyclonal antiserum possesses an additional antibody population, whose antigenic recognition did not contain the chiral center. The upper and lower limits of quantification of the developed ELISA were defined as 362 and 3.62 ng S-ibuprofen/ml, respectively, based on a 90% confidence interval. PMID- 10798241 TI - Photobiological properties of nabumetone (4-[6-methoxy-2-naphthalenyl]-2 butanone), a novel non-steroidal anti-inflammatory and analgesic agent. AB - The photolability of nabumetone (NB, 1, 4-[6-methoxy-2-naphthalenyl]-2-butanone) and its photobiological properties were studied under aerobic and anaerobic conditions using a variety of in vitro phototoxicity assays: photohemolysis, photoperoxidation of linoleic acid, and photosensitized degradation of histidine and thymine. The photodegradation rate of NB in methanol and phosphate buffered saline (PBS) was enhanced under oxygenated media. NB was phototoxic in vitro. The photohemolysis rate was enhanced by deuterium oxide and inhibited by the presence of 1,4-diazabicyclo[2.2.2]octane (DABCO), butylated hydroxyanisole (BHA), sodium azide (NaN3) and reduced gluthathione (GSH). The induced photoperoxidation of linoleic acid was inhibited significantly by sodium azide and reduced gluthathione. Histidine and thymine were photodegraded by a photosensitized reaction induced by NB. A mechanism involving singlet oxygen, radicals and photoproducts is suggested for the observed photoxicity. PMID- 10798243 TI - [Important pharmaceutical-chemical characteristics of the central muscle relaxant chlormezanone]. AB - The enantinomers of chlormezanone (1) may be achieved by enantioselective HPLC separation with a yield of 98% using a OD-Daicel column. Both enantiomers bind to human serum albumin (HSA) at pH 7.4 to a range of 11-12%. Binding to the globuline fractions is much less (2-4%, equilibrium dialysis, validation by ultrafiltration). It could be demonstrated by means of 1H-NMR spectroscopy that 1 binds to HSA with the benzene ring as well as with the thiazanone ring. The velocity of racemization could be measured for the first time using a BSA column. The enantiomers undergo racemization at pH 7.4 and 37 degrees C with a halflife of approx. 20.5 h. PMID- 10798244 TI - Studies on vaginal bioadhesive tablets of acyclovir. AB - Bioadhesive vaginal tablets were prepared using poly(acrylic acid) (PAA); Methylcellulose (MC), carboxymethyl cellulose (CMC), hydroxypropyl cellulose (HPC) and hydroxypropylmethyl cellulose (HPMC) as bioadhesive polymers in different concentrations and acyclovir as drug by direct compression technique (DCT) and wet granulation technique (WGT). Physical tests were applied to the tablets. The swelling behavior of vaginal tablets in distilled water, lactic solution and cow vagina, acyclovir release rate in lactic solution and bioadhesion to vaginal mucosa in cow vagina, in situ, were investigated. Swelling of the tablets containing HPC, CMC and MC was very rapid and caused disintegration of the tablets. The swelling behaviour of the tablets containing HPMC lasted 6 h in lactic solution. The force (N) necessary to detach the tablets from the vaginal tissue was found to depend on concentration and type of the bioadhesive polymer. The tablets containing HPMC needed the most detachment force. PMID- 10798245 TI - Predicting color kinetics during red Asian ginseng (Panax ginseng) preparation. AB - A red Asian ginseng preparation was prepared as follows: fresh Asian ginseng (Panax ginseng) roots were first steamed for half an hour to several hours, and then the steamed roots were dried into died roots (red Asian ginseng, 10% moisture content, on a dry basis). During steaming, the color of ginseng (white) turned yellow and brown during subsequent steaming. Color is an important index for grading red Asian ginseng. In this study, the effects of drying time and temperature on the surface color formation (L, a/b) of the Asian ginseng, and the color formation (L, a/b) of the red Asian ginseng powder were investigated. The value of L decreased, while the value of a/b increased as drying proceeded; the value of L was slightly influenced by the drying temperature, but the value of a/b was markedly influenced. With respect to the color of the final product (red ginseng), the value of L was slightly influenced by steaming time and temperature, while the value of a/b was increased as steaming time and drying temperature increased. Based on the nth-order rate equation and Arrhenius equation, a kinetic model for describing these effects was established, and the results were satisfactorily fitting. PMID- 10798246 TI - Charge-transfer chromatographic study of the interaction of antibiotics with cetyltrimethylammonium bromide. AB - The interaction of 20 antibiotics with the cationic surfactant cetyltrimethylammonium bromide (CTAB) was studied by charge-transfer reversed phase chromatography carried out on impregnated alumina layers using water methanol mixtures as eluents. The lipophilicity and specific hydrophobic surface area of antibiotics and the relative strength of their interaction with CTAB was calculated. CTAB interacted with 10 antibiotics the antibiotic--CTAB complex generally being more hydrophilic than the uncomplexed molecule. The relative strength of interaction depended considerably on the molecular structure of the antibiotics. Significant linear correlation was found between the lipophilicity of antibiotics and their capacity to interact with CTAB indicating the involvement of hydrophobic forces in the interaction. PMID- 10798247 TI - Cytotoxicity of rhenium(I) alkoxo and hydroxo carbonyl complexes in murine and human tumor cells. AB - The rhenium(I) alkoxo/hydroxo carbonyl complexes were shown to be very potent in suspended tumor cell lines in suppressing growth but were more selective in inhibiting the growth of cultures from solid tumors. Their mode of action in L1210 lymphoid leukemia cells indicated that they were not alkylating agents but interfered with nucleic acid metabolism at multiple enzyme sites, e.g. dihydrofolate reductase, PRPP-amido transferase, thymidine kinase, with DNA strand scission after 60 min incubation. These compounds did not function mechanistically exclusively as cisplatin derivatives causing intrastrand linkages of DNA but rather they mimicked the metal complexes of aminecarboxyboranes, furan oximes, N-substituted thiosemicarbazones, trifluoromethyl borons and ferratricarbadecarbanyl complexes acting as antimetabolites. PMID- 10798248 TI - Analgesic principle from Abutilon indicum. AB - Bioactivity guided isolation of Abutilon indicum yielded eugenol [4-allyl-2 methoxyphenol], which was found to possess significant analgesic activity. At doses of 10, 30, and 50 mg/kg body weight, eugenol exhibited 21.30 (p < 0.05), 42.25 (p < 0.01) and 92.96% (p < 0.001) inhibition of acetic acid induced writhing in mice. At a dose of 50 mg/kg body weight, eugenol showed 33.40% (p < 0.05) prolongation of tail flicking time determined by the radiant heat method. PMID- 10798249 TI - Further quinoidal derivatives from Rubia cordifolia L. AB - Two new quinoidal dimers, a new naphthohydroquinone anhydride, a known naphthoic acid ester derivative and two known benzoic acid derivatives were isolated from the chloroform fraction of a chloroform-methanol (1:1) extract of Rubia cordifolia L. The identification of these compounds was based on spectral analysis. PMID- 10798250 TI - [Reaction of furosemide, quimethazine and hydrochlorothiazide with thymol and sodium hypochorite]. PMID- 10798251 TI - Determination of maprotiline hydrochloride in tablets by ion-pair extraction using bromthymol blue. PMID- 10798252 TI - Preparation and evaluation of benzyl benzoate-loaded poly-epsilon-caprolactone nanoparticles. PMID- 10798253 TI - Influence of pH change on drug release from rectal suppositories. PMID- 10798254 TI - Antifungal and antiprotozoal activities of saponins from Hedera colchica. PMID- 10798255 TI - Flavonoids of crataegus stevenii. PMID- 10798256 TI - Repression. I. PMID- 10798257 TI - Sigmund Freud and the Lubavitcher Rebbe. PMID- 10798258 TI - Self-disclosure and psychoanalytic meaning: a psychoanalytic fable. PMID- 10798259 TI - Psychic reality and psychic change: a therapeutic journey. PMID- 10798260 TI - Swords into plowshares: exploring the recovery ethics of destructive lives. PMID- 10798261 TI - Defense and resistance in the psychoanalytic process. AB - "Defense" and "resistance" are usually regarded as two separate and distinct concepts, the first belonging to psychic functioning in general and the second to therapy, yet this distinction is far from constant. Clinical observations indicate a frequent overlapping of these two mental functions or positions. A patient's behavior may convey the meaning of either or both concepts, and a position or response that one assumes to bespeak resistance may reveal defense instead. This state of affairs may be seen in the analysis of patients with a relatively well-integrated neurotic identity, but its main import is in the analysis of psychosomatic patients and those with severe personality disorders. Insufficient holding experiences during the infancy of these patients do not enable them to develop functional structuralization. Deficient mothering in the first stages of life also prevents the stable differentiation between defense against threats from within and resistance against influence from outside. PMID- 10798262 TI - [Rediscovery of therapeutic hypothermia for newborn infants]. PMID- 10798263 TI - [Premature rupture of fetal membranes at term: sequelae of conservative management. An analysis of a personal patient sample]. AB - Few procedures are less standardised than the procedure in case of pre-labour rupture of the membranes at term (PROM). We propose that management should be reviewed regularly on the basis of one's own data and be modified accordingly if necessary. For the duration of three months we analysed 400 pregnancies retrospectively. Patients with PROM were observed expectantly for 24 hours. If there were no spontaneous uterine contractions, labour was induced, depending on the degree of cervical dilatation. 10 percent of the cases studied had PROM. Of these a high proportion of 73 percent were primigravida, likewise 73 percent had an unripe cervix. The average time between PROM and delivery was 27 h. 50 percent of the babies were born 24 h after PROM. If delivery occurred more than 24 h after PROM, the rate of caesarean section (15 vs. 30 percent), the rate of forceps deliveries (11 vs. 20 percent), the rate of amnionitis (16 vs. 35 percent) and the number of admissions to the newborn-ICU (16 vs. 25 percent) almost doubled. The patients were examined vaginally relatively often prior to delivery (up to 18 times, with a mean of 8 times). We therefore recommend active management 6-8 h after PROM, should there be no onset of spontaneous uterine contractions. This is particularly beneficial to primigravida with an unripe cervix. PMID- 10798264 TI - [Effect of hypertensive pregnancy complications on neonatal outcome of growth retarded fetuses]. AB - OBJECTIVE: To determine the effect of hypertensive disorders in pregnancy on the neonatal outcome of growth restricted fetuses. There is conflicting data on the effect of hypertension during pregnancy on the incidence of neonatal respiratory distress syndrome (RDS) and intraventricular hemorrhage. Some studies report a lower incidence of RDS and intraventricular hemorrhage in infants of hypertensive mothers, whereas other studies report a similar or higher incidence in infants born to hypertensive mothers. STUDY DESIGN: We performed a retrospective analysis of 220 growth restricted fetuses born between January 1, 1996 to July 1, 1997 at the Department of Obstetrics and Gynecology of the University-Hospital at Homburg/Saar. Data were obtained by review of the medical records. Growth restricted infants born to preeclamptic women or women with HELLP syndrome were compared to growth restricted fetuses born to mothers without hypertensive disorders. RESULTS: Growth restricted fetuses born to hypertensive mothers had a significant lower birth weight (p < 0.05). The incidence of RDS in children born to hypertensive mothers was significantly higher (p < 0.05, p < 0.01) and they stayed significantly longer in the neonatal intensive care unit (p < 0.01). In contrast to infants born to mothers with HELLP syndrome (n = 7) there was no difference in the incidence of intraventricular hemorrhage, infection, sepsis, necrotizing enterocolitis or cardiac complications (arrhythmia, insufficiency) in case of preeclampsia (n = 68). The perinatal mortality of infants born to hypertensive mothers was significantly higher (p < 0.05, p < 0.01). CONCLUSION: This study does not support the contention that hypertensive disorders in pregnancy have a beneficial effect on the postnatal course of IUGR infants. PMID- 10798265 TI - [Life situation of parents of twins and triplets: a qualitative study]. AB - BACKGROUND: Starting in the middle of the 1980s, an increasing incidence of multiple pregnancies in Germany has been reported, with assisted reproduction mainly contributing to this increase. In the management of multiple pregnancies, the main focus is on optimising the obstetric care. The difficult socio-economic and psychosocial situation of parents of multiples, however, is often underestimated. METHODS: The present study examines the life situation of parents of twins and triplets using focus groups as a qualitative research method. Nineteen mothers and fathers of twins and triplets were interviewed in four focus groups. Main topics were the parental reaction to the diagnosis of multiple pregnancy, the first time at home with the multiples, special situations and wishes and ideas for a better care during multiple pregnancy. RESULTS: The diagnosis of multiple pregnancy was made at early gestational age, with the parental reactions being described between "happiness" and "shock". With this diagnosis, however, the future parents feel alone. Caring for the babies is leading to the limits of parent's ability to take stress. Marital problems and social isolation are discussed controversially. Exceptional situations such as illness of the mother require fast and un-bureaucratic solutions. CONCLUSIONS: The interviews suggest that special problems of multiple parenthood such as social isolation, marital and psychic problems are not necessarily related to higher order multiples or handicapped children. It appears to be essential to offer a more individual care for women expecting multiples, whereas the main resource for improving care is in providing contact and information possibilities. PMID- 10798267 TI - [Ultrasonically established cystic periventricular leukomalacia (PVL): incidence and associated factors in Switzerland 1995-1997]. AB - BACKGROUND: Cystic periventricular leukomalacia (PVL) is an ischemic brain lesion that mainly affects preterm infants and causes severe neurological damage. Diagnosis is made by cranial ultrasonography. Objectives of this study were to determine the incidence, to identify associated factors and to evaluate the frequency of neurological abnormality at discharge. PATIENTS AND METHODS: Infants with PVL in Switzerland were systematically registered (Swiss Pediatric Surveillance Unit, Swiss Neonatal Network) over three years (1995 to 1997). They were compared to a control group matched for gestational age. RESULTS: Over the three year period 40 infants with PVL defined as at least 2 cysts with diameter = 2 mm in the periventricular region were registered (35 of them were preterm babies). In comparison with the matched controls the infants with PVL had received significantly less frequently antenatal corticosteroids (44 vs 78%, Event Rate Ratio 0.57, 95% Confidence Interval 0.38-0.68), they had lower umbilical cord arterial pH and lower Apgar scores; there was a trend to arterial hypotonia and hypocapnia associated with PVL. The infants of the study group needed more often mechanical ventilation or nasal CPAP (92% versus 67%; ERR 1.38, CI 1.07-1.77) and had more often intracranial hemorrhage (39 versus 14%; ERR 2.8, CI 1.13-6.96). 56% of the infants with PVL were considered abnormal at the neurological examination at discharge compared to 28% in the control group (p < 0.02). CONCLUSION: The incidence of PVL in Switzerland is 1.2% for preterm infants with a birth weight less than 1500 g. Cranial ultrasonography on infants at risk for PVL is important because 44% of the infants with PVL didn't show neurologic abnormalities at discharge. PMID- 10798266 TI - [Competent parenting: expectations of young parents of counseling on pregnancy and labor]. AB - BACKGROUND: Are expecting parents prepared to change their lifestyle to benefit their unborn child? Do they see a need for more knowledge and competence to be good parents? What are the key expectations with respect to pregnancy, delivery conditions, and infant health care? To answer these questions, a study presented them to expecting and young parents in Germany. METHODS: 123 delivery units were selected to represent the 1120 delivery facilities of Germany identified in 1997 from all accessible sources. 109 participated, and of 7862 distributed questionnaires 5900 (75%) were filled-in completely by parents. RESULTS: Most parents (73%) expect information about pregnancy, birth and child health care in any instance. 25% would request information but only in problem situations. Preferred topics were prevention of diseases of the mother and her child (97%), intra-uterine development, preparation for delivery as well as breastfeeding, and nutrition of the infant (90% each). Although books had been the most frequent source of information, pediatricians, midwives, and gynecologists received the highest scores for satisfaction with their advice. The majority of expecting and new parents wished to get personal advice. For the choice of the delivery place, 59% had consulted friends and relatives, 43% their gynecologist. The most important criteria for their decision were high medical standards (95%), the hospital's attitude towards breast feeding and rooming-in (94% each), as well as the availability of neonatal care (89%). Also of prime interest were mother-child contact immediately after delivery (99%), and well-rested personnel (96%). CONCLUSIONS: Across social classes, most expecting and new parents see a need of a broad spectrum of topics in health information and education related to pregnancy, delivery and the expected infant. As far as delivery is concerned, most expect a perfect combination of modern medical care and safety on the one hand, and the undisturbed humane experience of the birth on the other. PMID- 10798268 TI - [Concentration of estrogen and progesterone receptors in the lower uterine segment at term labor]. AB - BACKGROUND: To determine cytosol estrogen and progesterone receptor concentrations in the human lower uterine segment in different stages of cervical dilatation during parturition at term. PATIENTS AND METHODS: Biopsy specimens of the lower uterine segment were obtained from 51 women undergoing non-elective cesarean section at term. The stage of cervical dilatation at the time of cesarean section was < 2 cm (N = 14), 2 to < 4 cm (N = 13), 4 to 6 cm (N = 11) or > 6 cm (N = 13). The cytosolic estrogen and progesterone receptor concentrations were determined by enzyme immunoassays. RESULTS: Median estrogen receptor concentrations at < 2 cm and at 2-< 4 cm cervical dilatation were 2.12 fmol/mg protein and 2.03 fmol/mg protein, respectively. After a significant drop at 4-6 cm cervical dilatation (median: 1.08 fmol/mg protein), estrogen receptor concentrations raised again at > 6 cm cervical dilatation (median: 2.00 fmol/mg protein). Median progesterone receptor concentration was 84.7 fmol/mg protein at < 2 cm cervical dilatation, diminished significantly at 2-< 4 cm cervical dilatation (36.6 fmol/mg protein) and increased at further cervical dilatation (4 6 cm: 75.7 fmol/mg protein; > 6 cm: 83.7 fmol/mg protein). CONCLUSIONS: These data suggest that predominantly a decrease in cytosolic concentration of the progesterone receptor at the beginning of the active phase of first stage labor may play a crucial role in the hormonal control of cervical dilatation during parturition at term. PMID- 10798269 TI - [Low incidence of HELLP syndrome before the 32nd week of pregnancy- discontinuation of the Munich study]. PMID- 10798270 TI - [Imaging techniques in cardiology: three-dimensional echocardiography]. AB - Three-dimensional echocardiography offers new opportunities for clinical cardiology and the solution of scientific questions. Data acquisition is possible using different techniques: (1) Realtime 3D echocardiography with matrix-array transducers is the most promising approach, but is still limited by several difficulties; (2) 3D reconstruction is based on a number of sequentially acquired 2D image planes (like in multiplane TEE), which are put together afterwards. There are 2 ways of data analysis. 1. Morphological analysis. Surface rendering of the endocardial border can create perspectives not achievable with conventional methods such as the "en face" view of atrial septal defects or the atrial view of the mitral valve. Prolapsing leaflets and the spatial relationship can be identified much easier than using 2D methods. In complex congenital heart disease 3D echo may provide better spatial orientation and easier communication with the cardiothoracic surgeons. 2. Quantitative analysis of volumes, masses, and surfaces is only possible after manual contour tracing with several cut planes generated from the 3D data set. This procedure is time consuming and limits the use in clinical routine, even though validation studies demonstrated that 3D echo determination of masses and volumes is superior to one- or two dimensional techniques which are based on geometric assumptions. Furthermore, quantitative 3D analysis has a unique pre- and postinterventional diagnostic potential. FUTURE PERSPECTIVES: Combination with color Doppler data may lead to a more precise quantitation of valve regurgitations. Improvements of hard- and software will allow faster acquisition, reconstruction, and quantitative analysis. Assessment of regional myocardial perfusion may be possible in combination with left heart contrast agents. CONCLUSION: 3D echocardiography allows perspectives not achievable conventionally and has a great potential for precise quantitative and reproducible analysis of cardiac morphology which overcomes the limitations of 2D echocardiography. PMID- 10798271 TI - Endothelial function and hemostasis. AB - The vascular endothelium influences not only the three classically interacting components of hemostasis: the vessel, the blood platelets and the clotting and fibrinolytic systems of plasma, but also the natural sequelae: inflammation and tissue repair. Two principal modes of endothelial behaviour may be differentiated, best defined as an anti- and a prothrombotic state. Under physiological conditions endothelium mediates vascular dilatation (formation of NO, PGI2, adenosine, hyperpolarizing factor), prevents platelet adhesion and activation (production of adenosine, NO and PGI2, removal of ADP), blocks thrombin formation (tissue factor pathway inhibitor, activation of protein C via thrombomodulin, activation of antithrombin III) and mitigates fibrin deposition (t- and scuplasminogen activator production). Adhesion and transmigration of inflammatory leukocytes are attenuated, e.g. by NO and IL-10, and oxygen radicals are efficiently scavenged (urate, NO, glutathione, SOD). When the endothelium is physically disrupted or functionally perturbed by postischemic reperfusion, acute and chronic inflammation, atherosclerosis, diabetes and chronic arterial hypertension, then completely opposing actions pertain. This prothrombotic, proinflammatory state is characterised by vaso-constriction, platelet and leukocyte activation and adhesion (externalization, expression and upregulation of von Willebrand factor, platelet activating factor, P-selectin, ICAM-1, IL-8, MCP-1, TNF alpha, etc.), promotion of thrombin formation, coagulation and fibrin deposition at the vascular wall (expression of tissue factor, PAI-1, phosphatidyl serine, etc.) and, in platelet-leukocyte coaggregates, additional inflammatory interactions via attachment of platelet CD40-ligand to endothelial, monocyte and B-cell CD40. Since thrombin formation and inflammatory stimulation set the stage for later tissue repair, complete abolition of such endothelial responses cannot be the goal of clinical interventions aimed at limiting procoagulatory, prothrombotic actions of a dysfunctional vascular endothelium. PMID- 10798272 TI - [Augmented reality in echocardiography. A new method of computer-assisted training and image processing using virtual and real three-dimensional data sets]. AB - Augmented reality (AR) applications link real with virtual image data, in order to increase their information content. In medicine they are especially useful for education and for supporting the interpretation of three-dimensional (3D) image data. Simulators are used to train risky or expensive procedures. In the AR application EchoCom2 a 3D surface model of the human heart is linked with echocardiographic volume data sets. The 3D echocardiographic data sets are registered with the heart model to synchronize it's temporal and spatial orientation. The heart model together with an animated ultrasound sector represents a reference scenario, which displays the currently selected cutting plane within the echocardiographic volume data set. Modifications of the cutting plane within the echocardiographic data are transferred simultaneously and in real time to the reference scenario. The AR application is used as a simulator to train two-dimensional echocardiographic examinations and as an orientation and navigation aid for the exploration of 3D echocardiographic data sets. Beginners in echocardiography have only a rudimentary conception of the spatial relationship between the actual ultrasound image and the 3D anatomy of the heart. They are unable to translate multiple two-dimensional slices into a coherent 3D mental image of the heart. In EchoCom2 the trainee can interactively explore the 3D heart model and the registered 3D echocardiographic data sets by the animated ultrasound sector, whose position is controlled by an electromagnetic orientation and position system (EPOS). The data from the EPOS are used to calculate the echocardiographic images that are analogue to the position of the animated ultrasound sector. EchoCom2 is also used to support the interpretation of 3D echocardiographic data sets. The analysis of 3D echocardiographic data has to be done during a post processing. Defining the exact position of a cutting plane within the volume is difficult due to the lack of a standardized representation, the independence of the cutting plane of any transducer position and the possibility to calculate an indefinite number of views. The simultaneous representation of the current cutting plane both in the volume data, and in the heart model enables the examiner ad hoc to recognize it's position and the visualized structures. PMID- 10798273 TI - [A standardized documentation structure for data documentation in echocardiography. Work Team on Standards and LV Function of the Work Group on Cardiovascular Ultrasound of the German Society of Cardiology, Heart and Circulation Research]. AB - Presently, there are no well-defined standards for documentation of echocardiographic studies. Nevertheless, standards are essential to provide comparability of data and to realize electronic communication, both essential for quality management in echocardiography. Therefore, the working group "Standards and LV function" of the German Society of Cardiology developed a consensus for documentation of echocardiographic studies. In the present paper this consensus is presented and illustrated by typical clinical examples. Additionally, a prototype of a user-oriented software based on this data set is presented. The complete data set for transesophageal and transthoracic echocardiography and the software prototype can be downloaded at http:@echo.ma.uni-heidelberg.de. PMID- 10798274 TI - [Improved endocardial border delineation during dobutamine stress echocardiography using the left heart contrast medium BY 963]. AB - The interpretation of induced wall motion abnormalities during dobutamine stress echocardiography is affected in the case of impaired image quality. In 48 consecutive patients (mean age 62 +/- 9 years, 32 males, 16 females) with suspected coronary disease undergoing coronary angiography, the transpulmonary contrast agent BY 963 was given i.v. as bolus during dobutamine stress echocardiography (10-40 micrograms kg min, plus max. 1 mg atropine) to analyze improvements in endocardial border delineation. For each of the 16 segments of the left ventricle, the endocardial border delineation was evaluated. Using BY 963 the average number of non-evaluable segments decreased by 58% from 5.2% to 2.2% at rest (p = 0.008) and by 56% from 5.9% to 2.6% at maximal stress (p = 0.003) as compared to the non-contrast study for all patients. In patients with impaired image quality, defined as at least 1 non-evaluable segment at rest without contrast enhancement (N = 14), the number of non-evaluable segments decreased from 19.2% to 8.2% (p = 0.004) at rest and from 19.2% to 9.6% (p = 0.006) at maximal stress. The greatest decrease of non-evaluable segments was seen in the lateral and anterior segments of the apical views (maximum of 80%). The improved endocardial border delineation resulted in an improved agreement between two observers in the interpretation of the dobutamine stress echocardiograms as positive or negative (kappa = 0.38 without contrast, kappa = 0.58 with contrast). Contrast application resulted in a slight improvement of diagnostic accuracy of dobutamine stress echocardiography in the detection of angiographically proven significant coronary artery disease. CONCLUSION: In patients with impaired endocardial border delineation the use of the echo contrast agent BY 963 reduces the number of non-evaluable segments. Improvement of endocardial delineation is greatest for lateral and anterior segments in the apical views. PMID- 10798275 TI - [Pseudoaneurysm after Bentall's procedure in acute aortic dissection type A: diagnosis using electron beam tomography]. AB - Pseudoaneurysms are a rare complication following replacement of the ascending aorta or aorto-coronary bypass surgery. We report a case with replacement of the aorta ascendens, the aortic valve, and venous aorto-coronary bypass grafting, in whom a pseudoaneurysm developed at the site of one proximal bypass anastomosis. For the preoperative diagnosis, an ultrafast-CT (Electron Beam Tomography, EBT) was done, as neither the pseudoaneurysm, nor the venous graft could be visualized by selective coronary angiography. PMID- 10798276 TI - [Congenital tubular supravalvular aortic stenosis with massive coronary artery dilatation in a 35-year-old man]. AB - Supravalvular aortic stenosis is a rare cause of left ventricular outflow obstruction in adults. It occurs as an isolated defect sporadically or on a hereditary basis with an autosomal dominant trait without further phenotypical anomalies, or as part of the Williams syndrome with mental retardation and multiple other anomalies. This lesion was proved to result from a defect of the elastin coding gene. Supravalvular aortic stenosis is frequently associated with cardiovascular defects, particularly of the peripheral pulmonary arteries, thoracic aorta, carotid, subclavian, and coronary arteries and the aortic valve. The coronary arteries are subject to an increased perfusion pressure leading to dilatation, tortuosity and accelerated arteriosclerosis. We give details of a 35 year-old patient in whom a previously asymptomatic supravalvular aortic stenosis is associated with an excessive dilatation of the right coronary artery and the left anterior descending coronary artery as well as an ostium stenosis of the left common carotid artery. The patient did not present any phenotypical anomalies of the Williams syndrome. PMID- 10798277 TI - [Regulation of the endothelial function and angiogenesis by vascular endothelial growth factor-A (VEGF-A]. AB - VEGF-A (vascular endothelial growth factor-A) is an endothelial-specific growth factor that stimulates endothelial function and angiogenesis. VEGF-A plays an important role during development of the vascular system, wound healing, vascularization of tumors, and for angiogenesis in ischemic tissues including the heart. VEGF-A stimulates many actions of endothelial cells including proliferation, migration, and nitric oxide release via binding to and activation of the two primarily endothelial-specific receptor-tyrosine kinases KDR and Flt 1. KDR and Flt-1 stimulate multiple signal transduction pathways in endothelial cells. This review provides an overview of the role of VEGF-A in the regulation of endothelial function, angiogenesis, and arteriogenesis with regard to activation of signal transduction pathways and their functional consequences in the endothelium. Moreover, this article discusses recent developments exploring the therapeutic potential of VEGF-A for treatment of cardiovascular diseases. PMID- 10798278 TI - [Endothelium and endogenous fibrinolysis]. AB - Reduced activity of the endogenous fibrinolytic system contributes to intramural deposition of microthrombi in atherogenesis and to intraluminal deposition of thrombi leading to acute complications of atherosclerosis such as acute coronary syndromes. Endogenous fibrinolytic activity is predominantly regulated by the plasminogen activator inhibitor type 1 (PAI-1). Increased activity of PAI-1 leading to reduced endogenous fibrinolytic activity has been identified as an important independent risk factor for cardiovascular disease. Vascular endothelial cells form a barrier between the circulating blood with its dynamic balance between ongoing thrombosis and fibrinolysis and the subendothelial layers of the vascular wall with their prothrombotic activity. In addition, endothelial cells synthesize and secrete substantial amounts of plasminogen activators and their inhibitor PAI-1. Thus, endothelium plays an important role in the regulation of endogenous fibrinolysis. After describing the components of the endogenous fibrinolytic system and its interactions, this review focuses on the impact on endogenous fibrinolysis by the renin angiotensin system, the kallikrein kinin system, and type 2 diabetes mellitus. Investigations using transgenic and knock-out animal models--the results of which are also summarized--have improved our understanding of the interaction between endogenous fibrinolysis and endothelium. In each section of the review therapeutic implications and potentials are discussed. PMID- 10798279 TI - [Physical activity and training in heart failure]. AB - The main symptoms of chronic heart failure are dyspnea and exercise intolerance. The pathophysiological basis of these symptoms is not simply the dysfunction of the heart, but a complex interaction of the central circulation, the peripheral vessels, the skeletal muscles, the ventilatory, and the endocrine system. In contrast to acute heart failure, where bedrest is known to be beneficial, prolonged limitation of physical activity can be detrimental in the chronic stage of the disease. Therefore, since the 1980s there have been several reports about heart failure patients participating in exercise programs. The results were encouraging: the physiological gains were impressive, and contrary to prior fears, in the great majority of reports no deterioration of the cardiac function could be observed. The net result of training in this condition is an improvement in exercise capacity in the range of the best pharmacological treatment. In detail, blood flow into the working muscle is increased, the ventilation for each given workload is reduced, the skeletal muscle overall function (including biochemical and histological aspects) is improved, the increased neurohormonal activity--especially of the sympathico-adrenergic system--will be normalized, and the patient's quality of life is significantly improved. Whether all of these beneficial aspects will result in improved survival is not yet proven, although one recent study provides some evidence in this direction. But even if today no definite answer to this question from a large, multicenter trial is available, application of exercise training for selected heart failure patients can be recommended, if adequate supervision is provided. PMID- 10798280 TI - [New milestones in the therapy of chronic heart failure with beta blockers]. PMID- 10798281 TI - Angiogenesis and arteriogenesis--not yet for prescription. AB - Angiogenesis is the growth of new vessels from preexisting vessels by sprouting and intussusception with ischemia being the major stimulus. Circulating endothelial precursor cells have recently been found to participate in this process. The remodeling of preexisting bridging collateral arterioles, i.e., arteriogenesis, should be a much more efficient mechanism to compensate for the gradual or intermittent occlusion of a major epicardial or peripheral artery. Arteriogenesis is associated with an active growth process. It is probably not dependent on ischemia but initiated by local hemodynamic and mechanical effects on the vessel that occur with increasing blood flow. A variety of growth factors that act not only by stimulating endothelial and smooth muscle cell proliferation and migration, as well as substances that increase recruitment and activation of monocytes have been demonstrated to stimulate angiogenesis and arteriogenesis. Several clinical phase I trials suggest the feasibility and short-term safety of treatment with growth factors or their genes. The VIVA trial, the only phase II trial that has been published in this field, employed VEGF165 by intracoronary infusion followed by several intravenous infusions and did not demonstrate any increase of exercise time or angina by VEGF over placebo. The strong sustained placebo effect was surprising. Concerns about the long-term exacerbation of angiogenesis-dependent pathologic processes, like malignant tumors, atherosclerotic plaque formation and proliferative retinopathies, will require careful follow-up. Pro-angiogenic and pro-arteriogenic therapies may need further sophistication before they enter clinical practice. PMID- 10798282 TI - [Legal aspects: nonbinding recommendations or jurisdiction upon medicine?]. AB - The freedom of therapy granted to doctors through the ages by the jurisdiction does not imply a preferred position of the doctor but lies in the interest of the patient: the doctor after thorough examination should carry out self-responsibly the therapy that is in the best interest of the patient if and as far as the patient agrees. Unfortunately, today the freedom of medical treatment is thoroughly and from multiple sides endangered. Danger derives on the one hand from the increasing influence of jurisdiction on medicine, and on the other hand, from the increasing specialization and the segregation of fields of expertise resulting therefore. Most dangerous is the multiple and subtle regulation on free medical practice of the profession by reforming the mandatory health insurance with quotas, positive lists (allowed prescriptions), fixed budgets, etc. That the doctors have substantially contribute to the erosion of freedom of therapy which also increases the risk of criminal and civil liability of the doctor by excessive support of the creation of guidelines in the recent past, is not generally acknowledged. The following should cast a little light on the missing conscience. PMID- 10798283 TI - Latest on HRT & breast cancer risk. PMID- 10798284 TI - Blood pressure: why the top number counts. PMID- 10798286 TI - The rationale for off-label prescriptions. PMID- 10798287 TI - Your first line of defense against colorectal cancer. PMID- 10798285 TI - Enjoying lifelong sexual vitality. PMID- 10798288 TI - Are microwave ovens, cell phones, and other such devices dangerous to people with pacemakers? PMID- 10798289 TI - Which of the new smoking cessation treatments is best? PMID- 10798290 TI - Affinity chromatography on immobilized "biomimetic" ligands. Synthesis, immobilization and chromatographic assessment of an immunoglobulin G-binding ligand. AB - A synthetic bifunctional ligand (22/8) comprising a triazine scaffold substituted with 3-aminophenol (22) and 4-amino-1-naphthol (8) has been designed, synthesised, characterised and immobilized on agarose beads to create a robust, highly selective affinity adsorbent for human immunoglobulin G (IgG). Scatchard analysis of the binding isotherm for IgG on immobilized 22/8 (90 micromol 22/8/g moist weight gel) indicated an affinity constant (Ka) of 1.4 x 10(5) M(-1) and a theoretical maximum capacity of 151.9 mg IgG/g moist weight gel. The adsorbent shows similar selectivity to immobilized protein A and binds IgG from a number of species. An apparent capacity of 51.9 mg human IgG/g moist weight gel was observed under the experimental conditions selected for adsorption. Human IgG was eluted with glycine-HCl buffer with a recovery of 67-69% and a purity of 97.3 99.2%, depending on the pH value of the buffer used for elution. Preparative chromatography of IgG from human plasma showed that under the specified conditions, 94.4% of plasma IgG was adsorbed and 60% subsequently eluted with a purity of 92.5%. The immobilized ligand was able to withstand incubation in 1 M NaOH for 7 days without loss of binding capacity for IgG. PMID- 10798291 TI - New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography. AB - A new approach for the identification of ligands for the purification of pharmaceutical proteins by affinity chromatography is described. The technique involves four steps. Selection of an appropriate site on the target protein, design of a complementary ligand compatible with the three-dimensional structure of the site, construction of a limited solid-phase combinatorial library of near neighbour ligands and solution synthesis of the hit ligand, immobilisation, optimisation and application of the adsorbent for the purification of the target protein. This strategy is exemplified by the purification of a recombinant human insulin precursor (MI3) from a crude fermentation broth of Saccharomyces cerevisiae. PMID- 10798292 TI - Normal values and age-dependent changes in GTP cyclohydrolase I activity in stimulated mononuclear blood cells measured by high-performance liquid chromatography. AB - GTP cyclohydrolase I (GCH1) activity in phytohemaglutinin (PHA)-stimulated mononuclear blood cells (MBCs) is a useful clinical marker for diagnosis of tetrahydrobiopterin (BH4)-related genetic disorders such as recessively inherited GCH1 deficiency and dominantly inherited dopa-responsive dystonia (Segawa's disease). Since the assay is complex, including isolation of MBCs from blood, stimulation of MBCs by PHA under culture, isolation of the protein fraction from the PHA-stimulated MBCs, and the subsequent activity measurement, the reproducibility is problematic in its application to clinical study. We established a sensitive and reproducible method by high-performance liquid chromatography with fluorescence detection for clinical assay of GCH1 in PHA stimulated MBCs, and measured the normal values of 91 healthy males and females of various ages (1-74 years). The mean normal values were 19.1+/-0.9 pmol/mg protein per h (mean+/-S.E., n=91). There were no significant differences between males and females. The activity tends to be higher in the first decade and to be decreased from the second to third decade and becomes almost stable from the third decade. PMID- 10798293 TI - Simultaneous determination of the HIV protease inhibitors indinavir, amprenavir, saquinavir, ritonavir, nelfinavir and the non-nucleoside reverse transcriptase inhibitor efavirenz by high-performance liquid chromatography after solid-phase extraction. AB - As part of an on-going study on the suitability of a formal therapeutic drug monitoring (TDM) of antiviral drugs for improving the management of HIV infection, a high-performance liquid chromatography method has been developed to quantify simultaneously in plasma five HIV protease inhibitors (PIs) (i.e., indinavir, amprenavir, saquinavir, ritonavir, nelfinavir) and the novel non nucleoside reverse transcriptase inhibitor efavirenz. After viral inactivation by heat (60 degrees C for 60 min), plasma (600 microl), with clozapine added as internal standard, is diluted 1:1 with phosphate buffer, pH 7 and subjected to a solid-phase extraction on a C18 cartridge. Matrix components are eliminated with 2 x 500 microl of a solution of 0.1% H3PO4 neutralised with NaOH to pH 7. PIs and efavirenz are eluted with 3 x 500 microl MeOH. The resulting eluate is evaporated under nitrogen at room temperature and is reconstituted in 100 microl 50% MeOH. A 40-microl volume is subjected to HPLC analysis onto a Nucleosil 100, 5 microm C18 AB column, using a gradient elution of MeCN and phosphate buffer adjusted to pH 5.15 and containing 0.02% sodium heptanesulfonate: 15:85 at 0 min-->30:70 at 2 min-->32:68 at 8 min-->42:58 at 18 min-->46:54 at 34 min, followed by column cleaning with MeCN-buffer, pH 5.15 (90:10), onto which 0.3% AcOH is added. Clozapine, indinavir, amprenavir, saquinavir, ritonavir, efavirenz and nelfinavir are detected by UV at 201 nm at a retention time of 8.2, 13.0, 16.3, 21.5, 26.5, 28.7 and 31.9 min, respectively. The total run time for a single analysis is 47 min, including the washing-out and reequilibration steps. The calibration curves are linear over the range 100-10,000 ng/ml. The absolute recovery of PIs/efavirenz is always higher than 88%. The method is precise with mean inter day relative standard deviations within 2.5-9.8% and accurate (range of inter-day deviations -4.6 to +4.3%). The in vitro stability of plasma spiked with PIs/efavirenz at 750, 3000 and 9000 ng/ml has been studied at room temperature, 20 degrees C and +60 degrees C. The method has been validated and is currently applied to the monitoring of PIs and efavirenz in HIV patients. This HPLC assay may help clinicians confronted to questionable compliance, side effects or treatment failure in elucidating whether patients are exposed to adequate circulating drug levels. The availability of such an assay represents an essential step in elucidating the utility of a formal TDM for the optimal follow up of HIV patients. PMID- 10798294 TI - Development of predictive retention-activity relationship models of non-steroidal anti-inflammatory drugs by micellar liquid chromatography: comparison with immobilized artificial membrane columns. AB - The predictive and interpretative capability of quantitative chromatographic retention-biological activity models is supported by the fact that under adequate experimental conditions the solute partitioning into chromatographic system can emulate the solute partitioning into lipid bilayers of biological membranes, which is the basis for drug and metabolite uptake, passive transport across membranes and bioaccumulation. The use of micellar solutions of Brij35 as mobile phases in reversed-phase liquid chromatography has proven to be valid to predict some biological activities of different kinds of drugs. In this study, quantitative retention-activity relationship (QRAR) models to describe some of the biological activities and pharmacokinetic properties of non-steroidal anti inflammatory drugs (NSAIDs) with predictive and interpretative ability are obtained. These models are compared with those obtained using immobilized artificial membrane (IAM) column data taken from the literature. For NSAIDs, the statistical characteristics of the micellar liquid chromatography (MLC) QRAR models were better than or at least comparable to those of the IAM-QRAR models. PMID- 10798295 TI - Determination of the cephalosporin antibiotic cephradine in human plasma by high performance liquid chromatography with ultraviolet detection. AB - A simple and sensitive high-performance liquid chromatographic method, for the determination of cephradine in human plasma samples has been developed and validated. Cephradine and cephaloridine (internal standard) were extracted from human plasma by perchloric acid protein precipitation followed by centrifugation. Aliquots of the extracts were analysed by reversed-phase high-performance liquid chromatography (HPLC) utilising a polymeric reversed-phase PLRP-S column, followed by ultraviolet detection at 260 nm. The method has a working dynamic range from 0.2 to 30.0 microg/ml from 200 microl human plasma. The precision of the method at 0.2 microg/ml was 4.9% (intra-assay) and negligible (inter-assay) as calculated by one-way analysis of variance and the accuracy of the method at 0.2 microg/ml was -4.1% in terms of percentage bias. This method has been successfully applied to clinical studies including an oral bioequivalence study comparing the pharmacokinetics of 500 mg tablets of Kefdrin with 500 mg tablets of Velosef in healthy human volunteers. PMID- 10798296 TI - High-performance liquid chromatographic determination of p-aminohippuric acid and iothalamate in human serum and urine: comparison of two sample preparation methods. AB - A high-performance liquid chromatography method applied to determine p aminohippuric acid (PAH) and iothalamate (IOT) in serum and urine samples of patients was evaluated according to recovery, reproducibility and linearity utilizing narrow-bore columns. The mobile phase consisted of 0.15 M sodium dihydrogenphosphate with 1.2 mM tetrabutylammonium sulphate, the pH was adjusted to pH 4.6, acetonitrile was added to a final ratio of 95:5 (v/v), the flow-rate was set at 0.3 ml/min. The separation was achieved on a ODS Hypersil column (200 x 2.1 mm I.D.). The UV detector was set at 254 nm. PAH and IOT are used for evaluation of kidney function [effective renal plasma flow (ERPF) and glomerular filtration rate (GFR)]). Under the described chromatographic conditions two sample preparation techniques, ultrafiltration and acetonitrile precipitation were compared. The results demonstrate the accuracy of both methods in evaluation of ERPF and GFR. Due to its cost-effectiveness we recommend the acetonitrile precipitation method in clinical routine. PMID- 10798297 TI - Free malondialdehyde determination in rat brain tissue by capillary zone electrophoresis: evaluation of two protein removal procedures. AB - Two procedures for the determination of underivatised, free malondialdehyde in rat brain tissue have been evaluated. Both procedures are based on capillary zone electrophoresis (CZE) and UV detection at 267 nm and differ only with respect to the protein removal step, for which ultrafiltration or precipitation with acetonitrile have been employed. The total analytical processes include sample homogenisation, addition of antioxidant, protein removal, and separation and detection in the CZE system, and take less than 20 min. The CZE buffer consists of 10 mM borax and 0.5 mM CTAB at pH 9.3. The malondialdehyde peak reaches the detector about 3 min after injection as one of the very first peaks in the electropherogram. The limit of detection (3 S/N) is 0.2 microM, corresponding to 4 fmol for an injection volume of 20 nl. The method is fast, reproducible and has a large linear range, spanning 0-200 microM. PMID- 10798298 TI - Novel and sensitive method for the detection of anandamide by the use of its dansyl derivative. AB - The dansyl ester of anandamide was prepared and showed intense fluorescence on silica gel thin-layer chromatography when viewed under long-wavelength ultraviolet light (detection limit, 15 fmol). A high-performance liquid chromatography method for the quantitation of anandamide was developed using a C18 column (250 x 4.6 mm) with gradient elution (1% acetic acid-methanol) and detection at 255 nm. The method was applied to the measurement of anandamide in media from cultured hepatocytes. Sample preparation involved extraction with a C18 cartridge, derivatization with dansyl chloride, thin-layer chromatography, and quantitation. The detection limit in hepatocyte media is 4.3 nmol at a signal to-noise ratio of three. PMID- 10798299 TI - Separation and quantification of cholesterol and major phospholipid classes in human semen by high-performance liquid chromatography and light-scattering detection. AB - A high-performance liquid chromatographic method coupled with light-scattering detection for the separate and accurate quantification of cholesterol and main phospholipid classes was applied to human spermatozoa and seminal plasma (SP). This method is based on normal-phase chromatography with silica gel as stationary phase and a ternary gradient with hexane, mixtures of chloroform-methanol and water as mobile phase. Lipids are separated with a good resolution and a high reproducibility. About 5 x 10(6) spermatozoa or 25 microl of seminal plasma are sufficient to accurate quantitative analysis of phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidycholine (PC), sphingomyelin (SM) and cholesterol. PC is the predominant phospholipid class in spermatozoa (102+/-8 nmol/10(8) spermatozoa) whereas SM is the major in the SP (163+/-6 nmol/ml). Both in spermatozoa and SP, PI is the minor class of the phospholipids (12+/-1 nmol/10(8) spermatozoa and 24+/-2 nmol/ml). In conclusion, this method offers interesting perspectives for analysis of sperm lipid composition in semen samples with low quantities of spermatozoa. PMID- 10798300 TI - Arsanilic acid-Sepharose chromatography of pyruvate kinase from KB cells. AB - In the present study, arsanical-based affinity chromatography for pyruvate kinase (PK) isolation was explored. p-Arsanilic acid (4-aminophenyl arsonic acid), which contains an arsonic acid moiety structurally similar to inorganic pentavalent arsenate, was conjugated to Sepharose 4B via its para-amino group to form an As(V)-Sepharose matrix. The cellular proteins from KB cells bound to arsonic acid moieties were eluted by 50 mM sodium arsenate in Tris-HCl buffer (50 mM, pH 7.6). A single protein band with a molecular mass of 58 kDa was shown on a sodium dodecyl sulfate-polyacrylamide gel. By immunoblotting, amino acid sequencing and enzymatic analysis, the sodium arsenate-eluted 58-kDa protein was demonstrated to be a human PK (type M2). By using this one-step As(V)-Sepharose chromatography, PK from KB cells was purified 35.4-fold with a specific activity of 153.15 U/mg protein in the presence of 6 mM fructose-1,6-biphosphate. Although PK was eluted from an As(V)-Sepharose column with sodium arsenate, PK activity was apparently inhibited by the used eluent system, but not by p-arsanilic acid, indicating a specific interaction of As(V) to PK. In summary, our results indicate that As(V) Sepharose can serve as a simple and efficient chromatographic support for PK purification from KB cells. PMID- 10798301 TI - Determination of the vitamin D analog EB 1089 (seocalcitol) in human and pig serum using liquid chromatography-tandem mass spectrometry. AB - A liquid chromatographic-tandem mass spectrometric assay in human and pig serum has been developed for quantitative analysis of EB 1089 (seocalcitol). EB 1089 is a novel vitamin D analog under development for the treatment of cancer. The analyte was extracted from serum after protein precipitation using an automated solid-phase extraction procedure involving both a reversed-phase and normal-phase procedure on a single C18 cartridge. The analytical chromatography was performed using a Symmetri C8 50x2.1 mm, 3.5 microm column. The mobile phase was a linear gradient from 75% to 99% methanol with a constant concentration of 2 mM ammonium acetate. EB 1089 and the internal standard [d6]-EB 1089 were detected by using MS MS. The ion source was operated in the positive electrospray ionisation (ESI) mode. The assay is specific, sensitive, and has a capacity of more than 100 samples per day, with a limit of quantitation of 10 pg ml(-1) for a 1.0-ml sample aliquot. It is now used for routine analysis in connection with pharmacokinetic studies in humans and toxicokinetic studies in pigs. PMID- 10798302 TI - Liquid chromatography with multichannel electrochemical detection for the determination of trans-resveratrol in rat blood utilizing an automated blood sampling device. AB - A sensitive and selective multichannel liquid chromatography-electrochemistry method was developed for the determination of the natural product trans resveratrol (resveratrol) in rat blood. After administration of resveratrol, blood samples were periodically collected by an automated blood sampling device. Resveratrol was extracted from 150 microl of diluted blood (blood and saline at a ratio of 1:1) with acetonitrile containing 1% of trichloroacetic acid. Chromatographic separation was achieved within 12 min using a C18 (100x2.0 mm) 3 microm column with a mobile phase containing 20 mM sodium acetate, 0.5 mM EDTA, pH 4.5 and 21% acetonitrile at a flow-rate of 0.4 ml/min. A multichannel detector with glassy carbon electrodes was used, which can control up to four working electrodes simultaneously with applied potentials of +800, 700, 600, 500 mV vs. Ag/AgCl. The limit of detection was 2 ng/ml at a signal-to-noise ratio of 3:1 and the limit of quantitation was 4 ng/ml. The linearity of the calibration curve was obtained over the analytical range of 5-1000 ng/ml. The intra- and interassay precision was in the range of 2.5-4.4% and 1.2-4.3%, respectively. Using this method it was possible to quantify blood concentration following a single dose of resveratrol to rats with good accuracy and precision. Thus the pharmacokinetic properties of resveratrol in rats can be examined for intraperitoneal, oral and intravenous dosing. PMID- 10798303 TI - Automated determination of reboxetine by high-performance liquid chromatography with column-switching and ultraviolet detection. AB - A fully automated method including column-switching and isocratic high performance liquid chromatography (HPLC) was developed for quantitative analysis of the new antidepressant reboxetine, a noradrenaline reuptake inhibitor. After serum injection into the HPLC system and on-line sample clean-up on a silica C8 (10x4.0 mm I.D.) clean-up column with an eluent consisting of 2.5% acetonitrile in deionized water, the chromatographic separation was performed on an analytical column (Lichrospher CN; 250x4.6 mm I.D.) with an eluent of acetonitrile-aqueous potassium phosphate buffer (0.008 M, pH 6.4) (50:50). The UV detector was set at 273 or 226 nm. The limit of quantification was about 15 ng/ml at 273 nm and about 4 ng/ml at 226 nm. The day-to-day relative standard deviation ranged between 2.7 and 6.7% with recovery rates > or = 90%. Linear regression analyses revealed correlation coefficients > 0.998. The method can be applied to therapeutic drug monitoring of reboxetine as well as pharmacokinetic studies. PMID- 10798304 TI - A conceptual proposal for the study of the quality of rehabilitation care. AB - PURPOSE: To propose a framework for the assessment and ongoing, continuous improvement of the quality of rehabilitation care. METHOD: The generally clarified and accepted approaches to the study of the quality of care are examined in view of the inherent characteristics of medical rehabilitation and the differences between it and other fields of medical practice. RESULTS: The process and outcome of care are distinct but complementary and their elements should, in rehabilitation, be used jointly and simultaneously as indicators of quality. CONCLUSIONS: Rehabilitation is of long duration and consists of various phases, each of them focusing on a different issue. It is proposed to monitor activities of rehabilitation professionals (process elements) that constitute the focus of care of a given phase and simultaneously to monitor relevant outcome elements attributable to the activities of the given phase. Comparison of monitored elements (the observed care) to their standards (the expected care) would enable assessment of the quality and its continuous improvement. PMID- 10798305 TI - Long-term consequences of spinal cord injury on social participation: the occurrence of handicap situations. AB - PURPOSE: Handicap situations in daily life of persons with Spinal Cord Injury (SCI) is rarely evaluated in spite of their impact on long-term health. The purpose of the present study was to identify the occurrence of potential handicap situations in individuals with SCI and to determine the potential associations between the level of social participation and some characteristics of the person. METHODS: Four hundred and eighty-two individuals completed a mailed questionnaire that comprised the 'assessment of life habits', a tool developed to assess social participation in persons with disabilities. RESULTS: Significant disruptions were particularly observed in home maintenance, participation in recreational and physical activities as well as in productive activities and the achievement of sexual relations. However, many individuals successfully achieved various social roles despite the presence of disabilities. No indications of a potential impact of premature ageing on the accomplishment of life habits were observed. CONCLUSIONS: The severity of injury seems to significantly increase the occurrence of handicap situations as the individuals with tetraplegia reported carrying out their life habits with much more difficulty or requiring more assistance than those with a less severe impairment. PMID- 10798306 TI - Time use of stroke patients in an intensive rehabilitation unit: a comparison between a Belgian and a Swiss setting. AB - PURPOSE: Functional improvement after stroke has been related to the intensity of treatment. The present study was set up to observe how stroke patients spend their time in a rehabilitation unit. METHOD: Behavioural mapping was performed throughout a full working day in a Belgian and Swiss stroke unit. RESULTS: Patients were most frequently involved in therapeutic activities, 28% of the day in Belgium and 45% in Switzerland. Physiotherapy accounted for the majority of the therapy time. The Belgian patients spent 27% of the day in their own room and Swiss patients 49% of the day. The most striking finding was that the Swiss patients spent nearly 1.5 hours per day more in therapy. CONCLUSIONS: Differences between the two settings could only partially be explained by more favourable patient-staff ratios in the Swiss setting. Autonomous practice, group therapy sessions and family involvement have to also be considered. PMID- 10798307 TI - Commentary on the scope for rehabilitation in severely disabled stroke patients. PMID- 10798308 TI - Rehabilitation after severe stroke--an enthusiastic approval and a cautionary note. PMID- 10798309 TI - The scope for rehabilitation in severely disabled stroke patients. PMID- 10798310 TI - The scope for rehabilitation in severely disabled stroke patients. PMID- 10798311 TI - Rehabilitation is more than functional recovery. PMID- 10798312 TI - Production-orientated education and training of the mentally disabled in sheltered employment (PIONIER). AB - PURPOSE: The development and realization of a production-orientated education and training system for mentally disabled people in sheltered employment in the course of the project PIONIER. It focuses on the metal and electronic branches of a workshop with a mentally disabled workforce in Alsdorf, Germany. METHODS: For the development of the modules an assessment tool called MELBA was used to determine the abilities of the disabled people and the requirements of the tasks. Methods for the collection of this information was: observation, questionnaires and documents. RESULTS: Three months after the introduction of educational measures the improvements of the capacity for learning and education of disabled persons was visible. In the same way the development of their self-reliance and social abilities was recorded. The metal working branch was capable of fulfilling every task with respect to quality issues and within set delivery times. The electro working branch was accomplished enough to create new workplaces and to get orders from the computer industry. CONCLUSIONS: By the use of a qualification system like PIONIER the tasks of the disabled employees have been enlarged and enriched. In order to cope with the increase competition sheltered workshops have to realize new ways of vocational training for mentally disabled employees. PMID- 10798313 TI - Spermatozoa of the 'primitive type' in Scutigerella (Myriapoda, Symphyla). AB - The myriapod class Symphyla is of interest in that insects generally are assumed to be derived from symphylan-like ancestors. Male Symphyla form spermatophores that are picked up by the female. Both euspematozoa and paraspermatozoa are formed. In spite of the mode of fertilization their euspermatozoa were found to be of a kind that is typical of aquatic animals, so called 'primitive spermatozoa;' these are characterized by a short sperm head with a bilayered acrosome, a midpiece containing a few unmodified mitochondria, and a 9 + 2 flagellum. Scutigerella are unique among terrestrial arthropods in having 'primitive spermatozoa'; and together with horseshoe crabs they are only the second case in Arthropoda. Two further sperm plesiomorphies not found in other myriapods or insects are (1) the presence of a cytoplasmic canal housing the proximal flagellum and (2) the existence of microtubular triplets in centrioles, one of which acts as a basal body. Symphyla and Diplopoda both have a striated structure in the center of the subacrosomal material. The paraspermatozoa lack acrosome and nucleus but have a prominent crystal, a single mitochondrion, and two membrane systems. The structure of Scutigerella euspermatozoa is consistent with Symphyla being close to the stem group of Myriapoda plus Insecta. PMID- 10798314 TI - Ultrastructural and karyotypic examination of in vitro produced bovine embryos developed in the sheep uterus. AB - This study examined whether development of bovine in vitro produced (IVP) blastocysts in the sheep uterus resulted in morphologically and karyotypically normal elongation stage bovine blastocysts. Seven day IVP bovine blastocysts, resulting from either in vitro maturation and fertilization, nuclear transfer (NT), or parthenogenic activation, were surgically transferred at the blastocyst stage into sheep uteri. Sheep were sacrificed after 7-9 days, and blastocysts were flushed from their uteri. One of each kind of IVP bovine blastocyst was recovered from sheep uteri for analysis by transmission electron microscopy, and nine NT blastocysts were used to establish cell cultures that were analysed for chromosome complement. TEM analysis of in vivo-derived elongation stage bovine and ovine blastocysts was done for comparative purposes. Most ultrastructural features of the 13-19 day blastocysts were similar to earlier stage blastocysts except that distinct alternative mitochondrial morphologies were found between epiblast and trophectoderm cells. Monociliated cells, presumably nodal cells, were observed in the bovine epiblast and hypoblast, and retrovirus-like particles were elaborated by cells in these same areas. Development in the sheep uterus of IVP bovine blastocysts resulted in the presence of crystalloid bodies in the trophectoderm cells, and apoptotic and necrotic cells were observed in the epiblast tissue. Thus, in vivo incubation in the sheep uterus allowed nearly normal development to the elongated blastocyst stage and may be useful for assessment of NT bovine blastocyst developmental competence. Cell cultures derived from the NT blastocysts had normal chromosome complements suggesting that activation by ionomycin and 6-dimethyl-aminopurine did not cause detrimental changes in ploidy in those blastocysts that developed. PMID- 10798315 TI - Secretion of brochosomes during the ontogenesis of a leafhopper, Oncometopia orbona (F.) (Insecta, Homoptera, Cicadellidae). AB - Secretion of brochosomes, intricately structured symmetrical protein-lipid particles produced in the specialized Malpighian tubules of the Cicadellidae, was studied during the ontogenesis of Oncometopia orbona (F.). Unlike most other cicadellids, O. orbona displays sexual dimorphism in the production of brochosomes. The Malpighian tubule secretory cells of nymphs, males and young females produce spherical brochosomes of 0.3-1.4 microm in diameter, extruded after molts through the hindgut and spread across the integument as a coating. In females with developing eggs the same cells switch to producing rod-like brochosomes of 3.5-11.0 microm in length, which are used to powder the oviposition sites. The transition involves a dramatic change in morphogenesis of the secretory particles. Both types of brochosomes derive from homogenous condensing Golgi granules and acquire their definitive structure as a result of growth, differentiation of the contents into a wall and a core, and development of a lattice of surface invaginations. Final stages of this process take place in Golgi-derived vacuoles. In the development of rod-like brochosomes the condensing granules coalesce into masses several times larger than nascent spherical type. These giant granules flatten, then become doughnut-shaped and finally break open into a pair of linear particles. Whether the chemical composition differs between the two types of brochosomes remains unknown. PMID- 10798316 TI - Different types of response to foreign antigens by leech leukocytes. AB - We used morphological and immunocytochemical approaches to characterize and to show the behavior of cells involved in leech inflammatory responses. Leeches were injected with bacterial lipopolysaccharide, fluoresceinated yeasts, sulfate spheres and ciliates (Protozoa). Shortly after injection, migrating cells appeared in the area of injection. The response of the cells occurred in relation to the injected micro or macro antigens. Each injection first provoked a migration of cells towards the non-self material. Afterwards, different responses (degranulation, phagocytosis, encapsulation, melanization) occurred. The migrating cells involved in these series of processes have a similar behavior and are characterized by CD markers of macrophages, NK cells and granulocytes, which are typical of many invertebrates and vertebrates. PMID- 10798317 TI - The digestive cells of the hepatopancreas in Aplysia depilans (Mollusca, Opisthobranchia): ultrastructural and cytochemical study. AB - Digestive cells are the most abundant cell type in the digestive diverticula of Aplysia depilans. These are tall columnar or club shaped cells, covered with microvilli on their apical surface. A large number of endocytic vesicles containing electron-dense substances can be found in the apical zone, but the presence of many heterolysosomes of large diameter is the main feature of these cells. Glycogen particles and some lipid droplets were also observed. Peroxisomes with a circular or oval profile were common, but crystalline nucleoids were not detected in them, although a dense spot in the matrix was observed in a few cases. These organelles were strongly stained after cytochemical detection of catalase activity. The Golgi stacks are formed by 4 or 5 cisternae, with dilated zones containing electron dense material. Arylsulphatase activity was detected in the Golgi stacks and also in lysosomes. Cells almost entirely occupied by a very large vacuole containing a residual dense mass seem to be digestive cells in advanced stages of maturation. The observation of semithin and ultrathin sections indicates that these very large vacuoles are the result of a fusion among the smaller lysosomes. Some images suggest that the content of these large vacuoles is extruded into the lumen of the digestive diverticula. PMID- 10798318 TI - An immunohistochemical study of the insulin-, glucagon- and somatostatin immunoreactive cells in the developing pancreas of the chicken embryo. AB - The distributions and relative frequencies of insulin-, glucagon- and somatostatin-immunoreactive cells were studied in dorsal, ventral, third and splenic lobes of developing chicken pancreas during embryonic periods (10 days of incubation to hatching) by immunohistochemical methods. The regions of pancreas were subdivided into three regions: exocrine, light and dark islet. Round, oval and spherical shaped immunoreactive cells were detected in all four lobes. According to developmental stages, the types of lobes and the regions of pancreas showed various distributions and relative frequencies. In the splenic lobes, insulin, glucagon and somatostatin-immunoreactive cells were detected in exocrine, dark islet and light islet from time differentiation of splenic lobes, 13 days of incubation. The insulin- and somatostatin-immunoreactive cells of the third lobes were detected in exocrine and light islets from 10 days of incubation, and in dark islets from 15 and 11 days of incubation respectively. Glucagon-immunoreactive cells were detected in exocrine, dark and light islets from 16, 11 and 19 days of incubation respectively. These immunoreactive cells of the ventral lobes were detected in exocrine and light islets. However, dark islets were not found in this lobe. Insulin-immunoreactive cells were demonstrated from 10 days of incubation in these two regions. Glucagon immunoreactive cells were detected from 17 days of incubation in exocrine and 16 days of incubation in the light islets. Somatostatin-immunoreactive cells were demonstrated from 11 days of incubation in exocrine and 14 days of incubation in the light islets. In the dorsal lobes, insulin-immunoreactive cells were demonstrated in exocrine, dark and light islets from 12, 14, and 13 days of incubation, respectively. Glucagon- and somatostatin-immunoreactive cells were detected in dark and light islets from 13 and 14 days of incubation, respectively. Glucagon- and somatostatin-immunoreactive cells were demonstrated from 10 and 11 days of incubation in exocrine respectively. Generally, insulin immunoreactive cells were increased in light islets but decreased in light islets with developmental stages. However, glucagon-immunoreactive cells were decreased in light islets but increased in dark islets. In addition, somatostatin immunoreactive cells showed the same frequencies in light and dark islets with developmental stages except exocrine which increased with developmental stages. PMID- 10798320 TI - The Harderian gland of the Dhub lizard Uromastyx microlepis of the Kuwaiti desert: an ultrastructural approach. AB - Harderian glands exist in the orbits of most terrestrial vertebrates. The basic function of the gland is the lubrication of the eye. The present study was carried out to shed some light on the ultrastructure of the still enigmatic Harderian gland of the lizard Uromastyx microlepis, a common species in Kuwait and other Gulf areas. The Harderian gland of Uromastyx microlepis is well developed, relatively large in size and lingual in shape. The epithelial cells of the anterior part of the gland are characterized by the presence of membrane bound granules of almost homogeneous consistency. These secretory granules are gathered in compartments and separated by membranes and stacks of granular endoplasmic reticulum (GER). Most of the lumina were empty. Moderate amounts of GER, free ribosomes and pleiomorphic mitochondria were observed in the perinuclear area of the epithelial cells. The medial and caudal parts of the gland were rich in special secretory granules, GER, free ribosomes and pleiomorphic mitochondria. The anterior part of the gland could represent the future lacrimal gland of mammals. A network of myoepithelial cells was recognized around the gland tubules. While no melanocytes or lymphocytes were observed in the scarce interstitial tissue, macrophages, that might have an immune function in the gland, were observed. PMID- 10798319 TI - Apoptosis of mouse embryonic stem cells induced by single cell suspension. AB - Embryonic stem cells (ES cells) are pluripotential, and are therefore used to construct gene knock-out mice. We found that the apoptosis of mouse ES cells was induced when the cells were dispersed as single cells, whereas this process was suppressed when they proliferated in aggregates. The apoptosis of ES cells was repressed when the cells were cultured on feeders prepared from STO cells, a cell line established from embryonic fibroblasts. Culture supernatants from STO cells did not block the apoptosis of ES cells, which suggests that a direct interaction between ES cells and STO cells is required for the suppression of apoptosis. The viability of ES cells examined by the trypan blue exclusion test or by the MTT ((3-4,5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assay decreased dramatically when the cells were dispersed in phosphate-buffered saline PBS. Cellular activity was restored by the addition of culture medium for ES cells. Glucose in the medium was found to be a major factor responsible for the restoration. Amino acids also restored the decrease in reduction of MTT. Suspension of the ES cells in PBS(-) caused leakage of the nucleosome into cytoplasm. Results indicate that the single cell suspension of ES cells leads to leakage of substrates for oxidative phosphorylation from the mitochondria, and that these cells finally become committed to apoptosis. PMID- 10798321 TI - Localization and fate of aluminium in the digestive gland of the freshwater snail Lymnaea stagnalis. AB - The digestive gland of the freshwater snail Lymnaea stagnalis, exposed to water containing an elevated concentration of aluminium at neutral pH for up to 30 days, followed by a 20 day recovery period, was examined by light and electron microscopy and X-ray microanalysis. Aluminium was localized in the yellow granules present in the digestive and excretory cells and in the green and small granules present in the digestive cells. More aluminium, silicon, phosphorus and sulphur were present in all three granule types from aluminium exposed snails. The number of yellow and green granules from the digestive gland of aluminium exposed snails showed a progressive increase over the experimental period compared to controls. The number and aluminium content of the granules is likely to reflect the role of the digestive gland as a 'sink' for accumulated aluminium. We propose that intracellular monomeric silica is involved in the detoxification of aqueous aluminium which at neutral pH is largely in the form of an insoluble polyhydroxide. The increased amounts of sulphur and phosphorus in the granules are likely to be part of a broad response to metal loading but probably do not play a significant role in the storage and detoxification of aluminium. PMID- 10798322 TI - Chromatin condensation during Scrobicularia plana spermiogenesis: a controlled and comparative enzymatic ultracytochemical study. AB - In Scrobicularia plana testis, a nuclear acid phosphatase (ACPase) activity was detected in mid and late spermatids with the improved Gomori-chloride procedure. Lead deposits were first observed in mid spermatids at focal points over condensed chromatin strands, increasing in density as chromatin further condensated. In late spermiogenesis, lead deposits became concentrated between chromatin aggregates, and after total DNA compaction were transfered to the nuclear periphery and then shed into the cytoplasm. The specificity of the nuclear ACPase was tested against different pH values (3.9, 7.2, 7.8, 9.0), substrates (TPP, IDP, TMP, p-NCS, ATP, GTP, AMP, ADP, AMP-PNP) and inhibitors (NaF, levamisole, Zn, vanadate, theophylline). To further specify the nature of this nuclear ACPase, other enzymes were comparatively studied at their optimal pH values and at pH 5.0: nucleoside-diphosphatase, thiamin-pyrophosphatase, inorganic trimetaphosphatase, lysosomal arylsulfatases A and B, ATPase, GTPase, 5'-nucleotidase, adenylate kinase, and adenylate cyclase. Several other controls were introduced to exclude artefactual deposits induced by lead ions and tissue molecules. The results showed that the enzyme has an optimal pH at 5.0, a high specific affinity for beta-GP, and is inhibited by NaF, which suggests that it behaves as a type B-ACPase, and all controls demonstrated the specificity of the enzymic activity. Because lead deposits were specifically and temporally associated with spermatid chromatin condensation, when DNA and RNA synthesis, histones, phosphoproteins and RNA molecules strongly decrease, it is possible to suggest that the nuclear ACPase could be associated with DNA processing during chromatin compaction or involved in the hydrolysis of 2' and 3' nucleotides resulting from nuclear RNase action during RNA degradation. PMID- 10798323 TI - Stimulation of rat cutaneous fibroblasts and their synthetic activity by implants of powdered nacre (mother of pearl). AB - The components of the cutaneous envelope, the epidermis and the dermis, change in response to aging or environmental stress factors. The fibroblasts involved in maintaining skin tone are the main targets. Nacre, mother of pearl, from Pinctada maxima, which can stimulate and regulate bone forming cells, was implanted in the dermis of rats to test its action on the skin fibroblasts. This report describes the effect of nacre on the skin fibroblast recruitment and physiological activity. It resulted in enhanced extracellular matrix synthesis and the production of components implicated in cell to cell adhesion and communication (such as decorine) and in tissue regeneration (type I and type III collagens). The nacre implant produced a well vascularized tissue. The physiological conditions in the region around the implant are thus those required for the positive interactions between the dermis and epidermis which are fundamental for the physiological function of the skin. PMID- 10798324 TI - Peroxisomes and intracellular cholesterol trafficking in adult rat Leydig cells following Luteinizing hormone stimulation. AB - The present study was designed to explore the intracellular cholesterol trafficking in Leydig cells of adult rats following Luteinizing hormone (LH) injection. Histochemical techniques were used to demonstrate distribution of free cholesterol in Leydig cells of control and LH-injected rats. Two groups of sexually mature male Sprague Dawley rats (n=4/group) were used. Fifteen min following an injection of 200 microl of either saline (control) or luteinizing hormone (LH, 500 microg in saline) testes of rats were fixed by whole body perfusion using 0.5% glutaraldehyde and 4% paraformaldehyde in 0.1 M cacodylate buffer for 20 min. Fixed testes were cut into 3 mm3 and kept immersed in the fixative for further 15 min. Tissue cubes were then incubated at 37 degrees C in a medium containing cholesterol oxidase, 3,3'-diaminobenzidine tetrahydrochloride, horseradish peroxidase and dimethyl sulfoxide to histochemically localize free cholesterol in Leydig cells and processed for electron microscopy. Thin sections of these tissues were stained with aqueous uranyl acetate and lead citrate and examined with a Philips 201C electron microscope. In Leydig cells of control rats, free cholesterol was detected primarily in lipid droplets and plasma membrane. In the majority of Leydig cells, peroxisomes were unstained for free cholesterol, but occasionally few stained ones were present. Staining was not detected in mitochondria and smooth endoplasmic reticulum (SER) in Leydig cells of control rats. In LH-injected rats, lipid droplets, many peroxisomes, inner and outer mitochondrial membranes and some cisternae of SER in Leydig cells showed staining for free cholesterol. Fusion of Leydig cell peroxisomes with lipid droplets and mitochondria was also observed in the LH treated rats. These findings suggested that peroxisomes in adult rat Leydig cells participate in the intracellular cholesterol trafficking and delivery into mitochondria during LH stimulated steroidogenesis. Lipid droplets are used as one source for cholesterol for this process. PMID- 10798325 TI - Expression of Glut-4 and Glut-1 transporters in rat diaphragm muscle. AB - Glucose transporters (Gluts) are a family of membrane proteins responsible for the transport of glucose across cellular membranes. Generally, alterations of Gluts expression in limb skeletal muscle have been reported. However, the changes of Glut isoforms in respiratory muscle which contracts with a duty cycle have rarely been studied. This study was performed to evaluate at the light microscopy level the expression of Glut-4 and Glut-1 transporters in normal and denervated diaphragm by immunohistochemistry method with specific Gluts antibodies. The results showed Glut-4 immunoreactivity in both the cell periphery and the interior of myocytes. Glut-1 was also present in the cell border and in the interior of myocytes in control diaphragm. However, Glut-4 staining was stronger than Glut-1 staining in control diaphragm. In denervated hemidiaphragm, the Glut 4 immunolabelling decreased and Glut-1 increased. These data indicated that (1) Glut-4 and Glut-1 transporters were observed in diaphragm; and (2) there were alterations in the expression of both glucose transporters after denervation. These alterations in Glut isoforms after denervation may be associated with the removal of innervation itself, and/or may partly result from passive stretch imposed by inspiratory activation of the contralateral side. PMID- 10798326 TI - Chronic illicit drug use, health services utilization and the cost of medical care. AB - Few studies have examined the relationships between drug use, health services utilization and the cost of medical care for a community-based sample of drug users. The purpose of this study was to analyze recently collected data on chronic drug users (CDUs), CDUs who were also injecting drug users (IDUs) and non drug users (NDUs) to determine whether these groups exhibited differences in health services utilization and cost. In addition to descriptive analyses, these relationships were estimated with multivariate regression models. Data were collected in 1996 and 1997 through a standardized self-reported questionnaire administered to individuals who were recruited through community outreach activities in the USA. Annual differences in health services utilization between CDUs, IDUs and NDUs were estimated for three measures: number of times admitted to a hospital, number of outpatient visits and number of emergency room episodes. Results of this study indicate that CDUs and IDUs consumed significantly more inpatient and emergency care, but less outpatient services relative to NDUs. Analyses of total health care costs showed that CDUs and IDUs each generated about $1000 in excess services utilization per individual relative to NDUs. This research is the first study to compare differences in health services utilization and cost among out-of-treatment drug users relative to a matched group of non users in a community-based setting. The findings suggest that health care providers and managed care organizations should consider policies that promote more ambulatory care and discourage emergency room and inpatient care among drug users. Innovative and culturally acceptable approaches may be necessary to provide incentives without posing unusual financial hardship. PMID- 10798327 TI - Scaling back goals and recalibration of the affect system are processes in normal adaptive self-regulation: understanding 'response shift' phenomena. AB - This comment addresses a set of phenomena that have been labeled 'response shift'. We argue that many of these phenomena reflect recalibration of a goal seeking system and an affect-management system, both of which are involved in normal adaptive self-regulation. In brief, we hold that these systems act as feedback control mechanisms. The reference values for both systems continuously undergo gradual recalibration. Because in most circumstances the adjustments tend to occur with equivalent frequency in both directions, their cumulative effect is minimal. In situations of either unusually prolonged goal attainment (and overattainment) or unusually prolonged adversity (as occurs, e.g., with deteriorating health), the cumulative effect can be substantial. We believe that these latter recalibrations of reference value account for many response shift phenomena. Other such phenomena are accounted for by the principle of hierarchical organization among the self-regulatory goals that comprise the self. PMID- 10798328 TI - Participatory diagramming as a means to improve communication about sex in rural Zimbabwe: a pilot study. AB - It is increasingly recognised that unequal gender relations and poor communication between men and women about sexual matters, play a central role in the rapid transmission of HIV in sub-Saharan Africa. Analysis of how communication might practically be improved remains a critical area for investigation however. To this end a pilot study, conducted in January 1998 involving two all-female focus groups in two rural areas of Zimbabwe, explored the possibility of using 'participatory' methods and visual diagramming as a means to facilitate rural people's communication about issues of sexual health. While still provisional, the results hold considerable interest for future HIV/AIDS work in the region. As a research tool, diagramming provides richer, more nuanced data about sexual activity than wholly discursive focus groups. However, the technique also holds considerable potential for action research and positive interventions that seek to facilitate couples' more open communication and safer sexual decision making. The pilot established both that rural women were comfortable utilising the techniques and that they were prepared to use them to discuss the detail of their sex lives. The next and vital step, as participants themselves suggested, is to involve men in similar self-analytical activities. PMID- 10798329 TI - Is travel distance a barrier to veterans' use of VA hospitals for medical surgical care? AB - Lengthy travel distances may explain why relatively few veterans in the United States use VA hospitals for inpatient medical/surgical care. We used two approaches to distinguish the effect of distance on VA use from other factors such as access to alternatives and veterans' characteristics. The first approach describes how disparities in travel distance to the VA are related to other characteristics of geographic areas. The second approach involved a multivariate analysis of VA use in postal zip code areas (ZCAs). We used several sources of data to estimate the number of veterans who had priority access to the VA so that use rates could be estimated. Access to hospitals was characterized by estimated travel distance to inpatient providers that typically serve each ZCA. The results demonstrate that travel distance to the VA is variable, with veterans in rural areas traveling much farther for VA care than veterans in areas of high population density. However, Medicare recipients also travel farther in areas of low population density. In some areas veterans must travel lengthy distances for VA care because VA hospitals which were built over the past few decades are not located close to areas in which veterans reside in the 1990s. The disparities in travel distance suggest inequitable access to the VA. Use of the VA decreases with increases in travel distance only up to about 15 miles, after which use is relatively insensitive to further increases in distance. The multivariate analyses indicate that those over 65 are less sensitive to distance than younger veterans, even though those over 65 are Medicare eligible and therefore have inexpensive access to alternatives. The results suggest that proximity to a VA hospital is only one of many factors determining VA use. Further research is indicated to develop an appropriate response to the needs of the small but apparently dedicated group of VA users who are traveling very long distances to obtain VA care. PMID- 10798330 TI - Precursors of lethal violence: a death row sample. AB - A qualitative methodology based on the standards of criminal defense investigation was used to analyze the social and family histories of 16 men sentenced to death in California. Using a multisource cross-validation methodology, we assessed patterns of impairment, injury and deficit at each of four ecological levels: family, individual, community and social institutions. Investigation documented consistent and pervasive patterns of serious impairment, injury and deficit across the cases and levels. The men share numerous risk factors and few resiliency factors associated with violence. We found family violence in all 16 cases, including severe physical and/or sexual abuse in 14 cases; individual impairments in 16, including 14 with post-traumatic stress disorder, 13 with severe depression and 12 with histories of traumatic brain injury; community isolation and violence in 12; and institutional failure in 15, including 13 cases of severe physical and/or sexual abuse while in foster care or under state youth authority jurisdiction. Appropriate interventions might have made a difference in reducing lethal violence and its precursor conditions. PMID- 10798331 TI - Children and medicines: self-treatment of common illnesses among Luo schoolchildren in western Kenya. AB - In a rural area of western Kenya, primary schoolchildren's health seeking behaviour in response to common illnesses was investigated. 57 primary schoolchildren (age 11-17 years, median 13 years) were interviewed weekly about their health status and health seeking activities for 30 weeks. The children each experienced on average 25 illness episodes during this period. Most episodes could be categorised into 4 groups: 'cold', 'headache', 'abdominal complaints' and 'injuries'. One fifth (21%) of the illness episodes were serious enough to keep the children from school. In 28% of them, an adult was consulted, while 72% were not reported to an adult caretaker. Of the episodes without adult involvement, 81% remained untreated, while 19% were treated by the children themselves with either herbal or Western medicines. Of all the medicines taken by the children, two thirds were provided or facilitated by adults (assisted treatment) and one third taken by the children themselves without adult involvement (self-treatment). Among boys, the proportion of illnesses, which were self-treated increased with age from 12% in the youngest age group (< 13 years) to 34% in the oldest (> 14 years). In girls, the proportion of illnesses which were self-treated was consistently lower than among boys and remained constant around 9% for all age groups. The proportion of Western pharmaceuticals used for self-treatment increased with age from 44% in the youngest age group to 63% in the oldest (average 52% Western pharmaceuticals). Again, there were differences between boys and girls: among the youngest age group, boys were twice as likely to use pharmaceuticals than girls (62 versus 32% of the self-treatments, respectively) and in the oldest age group they were nearly three times more likely (75 versus 25%, respectively). These differences in self-treatment practices and choice of medicines between girls and boys may reflect the higher income potential of boys, who can earn money by fishing. Pharmaceuticals were generally preferred for the treatment of headache and fevers, or colds, while herbal remedies were the preferred choice for the treatment of abdominal complaints and wounds. The most commonly used pharmaceuticals were antimalarials (mainly chloroquine), painkillers and antipyretics (mainly aspirin and paracetamol), which were stocked in most small shops in the village at low prices and readily sold to children. Throughout primary school age Kenyan children are growing into a pluralistic medical practice, integrating Western pharmaceuticals into the local herbal medical system, and gradually become autonomous agents in their health care. PMID- 10798332 TI - Deviant insiders: medical acupuncturists in New Zealand. AB - Acupuncture gained considerable attention in anglophone countries in the 1970s. As part of that popularity many medical practitioners became interested in the therapy and learned acupuncture techniques. A number of studies have indicated that medical practitioners were able to take up the practice of acupuncture without threatening the cultural authority of medicine so long as they limited the scope of its practice and redefined acupuncture concepts in Western biomedical terms. These analyses tend to present the medical profession as monolithic and emphasize a dichotomous relationship between biomedicine and alternative therapies such as acupuncture. This study examines the ways in which acupuncture has been represented in different medical forums, suggesting that in order to understand the relationships between biomedicine and alternative medicine we need to be more aware of the changing nature of these representations and their dependency upon the context of the representation. Rather than emphasizing a duality between orthodox medicine and alternative medicine, it is argued here that there are pluralities of medical and healing worldviews. PMID- 10798333 TI - The socio-economic determinants of maternal health care utilization in Turkey. AB - The purpose of this study is to investigate the individual-, household- and community-level factors that affect women's use of maternal health care services in Turkey. The data used for the study come from the 1993 Turkey Demographic and Health Survey (TDHS), a nationally representative survey of ever married women 15 to 49 years of age. In order to assess the impact of socio-economic factors on maternal health care utilization, we use logistical regression techniques to estimate models of the prenatal care use and birth delivery assistance among women who have had at least one birth in the three years prior to the survey. Separate models are also estimated for urban and rural women. The results indicate that educational attainment, parity level, health insurance coverage, ethnicity, household wealth and geographic region are statistically significant factors that affect the use of health care services thought essential to reduce infant and child mortality rates. The results of the model are used to provide insights for both micro- and macro-level planning of maternal health service delivery. PMID- 10798334 TI - Women's health in relation with their family and work roles: France in the early 1990s. AB - In this paper, the health of women aged 30 to 49 years is analyzed according to the family and work roles which they exercise, based on the 1991-1992 French national health survey. Households are classified based on the amount of their material resources, and a variety of measures of health and of health-related behaviors are considered. Looking at each role separately, the 'healthy married', 'healthy mother' and 'healthy worker' effects are very obvious for almost all health measures, and higher household income per unit of consumption is clearly associated with better health of women. The role patterns of women are not evenly distributed across income levels: housewives and lone mothers are more common at the bottom and middle of the income scale than at the top, while working women without children, married or not, are much more common at the top. In health terms, more heterogeneity is attached to role patterns in the middle of the income scale than at either extreme. In the middle stratum, two groups of women stand out as being clearly disadvantaged in comparison with that of married women with children and a job: (1) lone mothers, particularly in terms of mental health conditions, malaise symptoms and health-related behaviour, and (2) housewives, particularly in terms of physical health conditions. At the bottom of the income scale, no significant disadvantage is found for housewives compared to married working mothers, yet their overall health pattern is somewhat negative. At the top of the income scale, married working women without children, as well as single women do feel more often than married working mothers that they suffer from handicap or discomfort. The findings are discussed in terms of role enhancement and role strain, health selection, the nature of the health disadvantage associated with specific role patterns, and the importance of the structural context in the role framework. PMID- 10798335 TI - Parenthood, gender and sickness absence. AB - It is well documented that women have generally higher morbidity rates than men. In line with this women are also more absent from work due to sickness. This paper considers one popular explanation of the morbidity difference in general and of the difference in sickness absence in particular, viz. that women to a greater extent than men are exposed to the 'double burden' of combining paid work with family obligations. We discuss theories of role overload and role conflict, which both assume that the combination of multiple roles may have negative health effects, as well theories of role enhancement, which assume positive health effects of multiple roles. Using two large Norwegian data sets, the relationship between the number of and the age of children on the one hand and sickness absence on the other is examined separately for men and women and for a number of theoretically interesting subpopulations of women defined in terms of marital status (also taking account of unmarried cohabitation), level of education, and working hours. Generally speaking the association between children and sickness absence is weak, particularly for married people of both genders. To the extent that married persons with children are more absent than married persons without children, this is largely due to respiratory conditions. The relationship between children and sickness absence is somewhat stronger for single, never married mothers, but not for single mothers who have been previously married or for women living in unmarried cohabitation. The findings thus provide little support for either role overload/conflict or role enhancement theories. The possibility that these effects are both present and counterbalancing each other or that they are confounded with uncontrolled selection effects can not, however, be ruled out. PMID- 10798336 TI - Interpretative repertoires of medication among the oldest-old. AB - The use of medical drugs is not founded on medical knowledge alone, but it is also dependent on lay logic and reasoning. This study set out to explore the views of the oldest-old on their medication. The data for the study came from narrative interviews with people aged 90 or over. Our aim was to look for different culturally shared interpretative repertoires used by the interviewees as they gave descriptions and accounts of their drug use and presented themselves as users of medical drugs. Three interpretative repertoires were identified. The moral repertoire stressed lay people's moral norms and presented them as morally acceptable and responsible users of drugs by explaining and minimizing. The patient repertoire was used by the respondents to show they had accepted the role of patient. The self-help repertoire was used by the respondents to emphasize that they had made their own choices in medical care despite the biomedical facts. These repertoires showed that not only the biomedical logic, but also other logics are valid in the everyday world where most medical drugs are used. A better understanding of cultural ideas of drug use would help to improve the care of older people. PMID- 10798337 TI - Technology, selfhood and physical disability. AB - Much has been written in recent times of the interface between technologies and the human body. The vast majority of this literature, however, focuses on a body that is assumed to be free of physical disability. This article seeks to address this lacuna by presenting findings from an exploratory study using in-depth interviews with fifteen people with physical disabilities living in the Australian city of Adelaide. The dominant research question was to explore the ways in which technologies contribute to the meanings and experiences of the lived body/self with disabilities. The data showed that the interviewees identified several technologies that they used as highly beneficial to allowing them to transcend some aspects of their disabilities. However, the interviewees also identified significant negative aspects to the use of some technologies. They noted that such technologies could serve to mark out people with disabilities as 'different' or 'lacking', acting as a barrier to the achievement and presentation of their preferred body/self. PMID- 10798338 TI - The power of place: space and time in women's and community health centres in South Australia. AB - This paper focuses on the importance of time and space in an Australian medical setting. It draws on research findings from a one year project that aimed to explore community perspectives of, and experiences of medical services in three South Australian women's and community health centres. Both qualitative and quantitative methods of data collection and analysis were used in order to address these objectives. A significant finding was the way in which participants described the organisation and experience of time and space in these centres and how this impacted on their health and well being and that of the community. In analysing these spatio-temporal dimensions and the underlying philosophical structures of women's and community health centres, this paper argues that experiences associated with space and time have a positive effect on health status by: diminishing barriers to health services, improving quality of care, increasing community participation, providing safe places for social interaction and strengthening people's sense of belonging or attachment to a particular community and place. Based on these findings, the authors conclude that the spatio-temporal dimensions of health care provision have empowering and positive impacts on a community's health, a significant finding that has implications for the maintenance and future funding of this style of health service. PMID- 10798340 TI - Cellulose and calcium lower the incidence of chemically-induced colon tumors in rats. AB - In a 30-week preliminary study and a follow-up 22-week study (2 x 2 factorial), dimethylhydrazine (DMH) injections effectively induced colon tumors in Fischer 344 rats. How this incidence of colon tumors might be affected by cellulose (preliminary study) or by calcium and folic acid (follow-up study) was examined. Cellulose in the diet appeared to provide some protection against DMH-induced colon tumors, but the protective effect of calcium was more evident; normal levels of calcium (500 mg per 100 g diet), but not of folic acid (0.1 mg per 100 g diet), provided protection against colon tumors. The effect due to calcium was observed whether viewed in terms of total number of tumors (p < 0.01) or number of tumors per tumor-bearing rat (p < 0.01). PMID- 10798341 TI - Variability in physicochemical and nutritional quality traits of parents, F2 and F3 generations of lentil crosses. AB - Ten physicochemical and nutritional quality traits (100-seed weight, seed volume, seed density, bulk density, hydration capacity, hydration index, swelling capacity, protein, tryptophan and energy value) were examined in the seed samples of parents, F2 and F3 generations of three microsperma x microsperma and two microsperma x macrosperma crosses of lentils. Significant variation for different traits was observed among the genotypes in both the groups and also among both types of crosses in F2 and F3 generations. Ranges, means and coefficients of variation (CV %) for various traits among parents, F2 and F3 generations of different crosses, along with the deviations between F2 and F3 generation means for various traits in different crosses are discussed. Also suggested is a breeding strategy for evolving lentil varieties with improved seed yield and nutritional quality parameters. PMID- 10798339 TI - Bioavailability of iron from four different local food plants in Jordan. AB - The bioavailability of iron from local plants (black cumin seeds, milk thistle seeds, sesame seeds and thyme leaves) was investigated. Apparent absorption of iron was calculated by subtracting fecal iron (using total collection of feces) from iron intake in Sprague-Dawley rats. Two trials of animal feeding were performed. Liver and serum concentrations of iron, and serum hemoglobin concentration were taken as response parameters for the bioavailability. Dry thyme was particularly rich in iron (117.2 mg/100 g dry matter) and milk thistle seeds in crude fiber (25 g/100 g dry matter). Rats fed the dry thyme diet or that supplemented with 5% egg white died but when the diet was supplemented with 10% egg white, the animals survived. Iron intake and apparent and total iron absorption were highest for the rats fed the dry thyme-egg white diet. Liver weights for the groups fed black cumin seeds and dry thyme were significantly higher (p<0.05) than those for the groups fed milk thistle and sesame seeds. Liver content of iron was highest for the animals fed black cumin seeds. Serum iron content increased significantly (p<0.05) for the animals fed black cumin seeds, and serum hemoglobin concentration increased significantly (p<0.05) in the groups fed milk thistle seed and black cumin seed diets, but decreased for the group fed the thyme-egg white diet. It is concluded from this study that iron was better utilized from black cumin seeds as indicated by liver storage of iron. On the other hand, thyme had the highest iron absorption but lowest utilization. PMID- 10798342 TI - Effect of extrusion process variables on physical and chemical properties of extruded oat products. AB - The purpose of this research was to study the effects of initial moisture levels and extrusion temperatures on bulk density, water absorption and water solubility indexes, viscosity, and color of extruded oat products. The dehulled grains were ground in a Brabender Quadrumat Senior mill and the coarse fraction, with higher amounts of crude protein, lipids, and dietary fiber content, were conditioned to moisture levels of 15.5-25.5% and extruded in a Brabender single-screw laboratory extruder. The water absorption index of extrudates were relatively low (4.16-6.35 g gel/g sample) but increased as the initial moisture of the raw material as well as the extrusion temperature was elevated. The water solubility index was inversely proportional to the extrusion temperature. Initial viscosity of the paste increased with the increase of raw material moisture and extrusion temperature, while the maximum viscosity (at a constant temperature) diminished with the increase of temperature. Products with lower values of L* (luminosity) and greater values of a* (red) and b* (yellow) were obtained at high moisture rates and at a 120 degrees C extrusion temperature. PMID- 10798343 TI - Nutrient and antinutrient composition of Amaranthus muricatus. AB - The chemical and nutritional composition of the aerial parts of Amaranthus muricatus was assessed. This is a wild plant growing well in arid and poor soil, which are not suitable for cereals. The material under study was boiled, dried and ground and then evaluated for chemical, nutritional and antinutritional properties. Protein concentration of the amaranth flour was 15.74 g/100 g and total dietary fiber content was high (53.81 g/100 g), with 79% coming from insoluble dietary fiber. The calcium content was 1533 mg/100 g and the Ca/P ratio (5.94) was markedly higher than the value suggested in the 1989 Recommended Dietary Allowances. Fifty-nine percent of the fatty acids present were unsaturated with linoleic acid accounting for 40% of the total fatty acid content. Nitrate, hemagglutinin, trypsin inhibitor and saponin contents were within the non-critical values. Net protein utilization (NPU), true digestibility (tD) and biological value (BV) measurements indicated a nitrogen gain of 74% that of casein. The results of this study indicate a nutritive potential for the A. muricatus leaves, therefore, domestication of this plant is suggested along with assessment of its chemical and nutritional properties. PMID- 10798344 TI - Caffeine and theobromine levels in selected Nigerian beverages. AB - Caffeine and theobromine contents (mg/g) were determined in samples of selected Nigerian beverage products. The beverages were cocoa (Milo, Bournvita, Rosevita and Enervita), coffee (Nescafe, Bongo, and Maxwell House decaffeinated) and tea (Lipton). The theobromine contents of samples of Milo, Bournvita, Rosevita, Enervita, Nescafe, Bongo, Maxwell House decaffeinated coffee and Lipton were 62.10+/-5.21, 64.80+/-6.72, 82.80+/-4.43, 80.37+/-6.80, 27.00+/-4.31, 14.67+/ 2.90, 23.46+/-3.13 and 12.60+/-1.52, respectively. The corresponding caffeine contents of these samples were 2.78+/-0.43 (Milo), 3.17+/-0.36 (Bournvita), 0.92+/-0.51 (Rosevita), 1.05+/-0.68 (Enervita), 93.66+/-8.91 (Nescafe), 6.47+/ 2.42 (Bongo), 37.22+/-5.34 (Lipton), and 0.21+/-0.11 (Maxwell House decaffeinated coffee). Semi-processed cocoa beverages (Rosevita and Enervita) had significantly (p < 0.05) higher levels of theobromine compared with the finished cocoas (Milo and Bournvita). Similarly, Nescafe contained significantly (p < 0.05) higher levels of caffeine compared to Maxwell House (decaffeinated coffee) and Bongo. Levels of caffeine in Lipton tea were moderate. PMID- 10798345 TI - Chemical analysis and nutritional assessment of Teramnus labialis (L.) Spreng. (Fabaceae). AB - Seeds of Teramnus labialis (L.) Spreng. (Fabaceae), used as food by Malayali tribals in the Kollihills of Salem District, Tamil Nadu in South India, were investigated for their nutritional value. These seeds were analyzed for proximate composition, total (true) seed proteins, seed protein fractions, amino acid composition, fatty acid composition, minerals and antinutritional factors. Crude protein, crude fat, ash, and nitrogen free extracts constituted 22.86%, 6.10%, 4.62%, and, 58.15%, respectively, of the seed weight. The caloric value of 100 g dry matter of seed material was 378.94 kcal. The essential amino acids lysine, leucine + isoleucine, arginine, valine and histidine were present in relatively large quantities. The unsaturated fatty acids constituted more than 60% of the crude fat. Concentrations of minerals such as potassium, magnesium, calcium, and, phosphorus were high. Antinutritional factors such as total free phenols, tannins, L-DOPA, hydrogen cyanide and phytic acid were present in minute quantities. These antinutritional factors are potentially eliminated using the conventional method of soaking the seeds in water, boiling with water and decanting prior to consumption. PMID- 10798346 TI - Chemical composition and selected functional properties of sweet orange (Citrus sinensis) seed flour. AB - Flour samples were prepared from dehulled and undehulled sweet orange (Citrus sinensis) seeds. The flour samples were evaluated for proximate composition, mineral content and selected functional properties. Proximate analysis showed a composition of 54.2% fat, 28.5% carbohydrate, 5.5% crude fiber, 3.1% crude protein and 2.5% ash for the dehulled orange seed flour (dry weight). Mineral analyses showed high levels of calcium and potassium in flour samples. Partially defatted and undefatted flour samples prepared from dehulled orange seeds had least gelation concentrations of 10 and 12% (w/v), respectively. Water absorption capacity for the defatted and undefatted dehulled flour samples were 240 and 220%, respectively. Defatting improved oil absorption capacity of the orange seed flour by 84%. Emulsion activity, emulsion stability and foaming capacity decreased following defatting of flour. Foam prepared from defatted flour was less stable than that from full-fat flour. Incorporation of NaCl up to 0.2 M improved the foaming capacity of orange seed flour. PMID- 10798347 TI - Traditional and possible technological uses of Digitaria exilis (acha) and Digitaria iburua (iburu): a review. AB - The focus of this article is the traditional and technological uses of two tropical cereal grains: Digitaria exilis (acha) and D. iburua (iburu); with emphasis on their carbohydrate and protein components. Some useful attributes of the grains emanating from present knowledge are discussed. The major classes of traditional foods from these grains are thick and thin porridges; steam cooked products, e.g. couscous; and nonalcoholic and alcoholic beverages. The proteins in these grains are not easily extractable; however, the digestibility of the proteins are better than those of sorghum and millet. The high levels of residue protein in them may have important functional properties. Technologically, acha and iburu can be utilized in ways similar to rice. The two grains require minimal processing due to grain size and location of constituents. Whole acha and iburu grains are now used for quick cooking, non-conventional food products including weaning foods of low bulk density and breakfast cereal with good fiber content. The grains could be used in a wide variety of other products. Cookies, crackers, and popcorn, made in an almost endless array of forms are examples. The breeding of acha and iburu cultivars with good kernel properties is critically important to their utilization potential. PMID- 10798348 TI - Possible nutritional implications of varietal influence on the 7S/11S seed globulin ratios in amaranth. AB - 7S/11S seed globulin ratios were determined for seven currently available Amaranthus hybrid lines i.e., Amaranthus K266, K283, K343, K432, K433, K436 and MT-3. Of the seven Amaranthus lines investigated, four lines had 7S/11S globulin ratios ranging from 0.47 to 0.81, while three lines were found to contain exclusively the 11S globulin form. In general, 7S globulins contained lower levels of essential amino acids such as tryptophan, methionine, lysine, histidine, phenylalanine, valine and isoleucine than the 11S globulins. PMID- 10798349 TI - Studies on the application of dynamic surface tensiometry of serum and cerebrospinal liquid for diagnostics and monitoring of treatment in patients who have rheumatic, neurological or oncological diseases. AB - Human biological liquids comprise various surfactants, which adsorb at liquid interfaces and lead to a variation in surface tension. The adsorption processes involving low molecular weight surfactants, proteins and phospholipids play a vital role in the physiological functions of the human organism, especially if large surfaces are involved (e.g., gas exchange in lungs, metabolism of kidneys, liver and brain). Dynamic surface tensiometric studies of biological liquids like serum and cerebrospinal fluid provide surrogate parameters that reflect surface tension phenomena. We provide dynamic surface tension data of serum and cerebrospinal fluid that were collected from healthy volunteers and patients with rheumatic, neurological or oncological diseases. Our studies indicate that dynamic surface tension data are helpful for diagnostic purposes and for monitoring of therapeutic interventions. PMID- 10798350 TI - Dynamics of protein and mixed protein/surfactant adsorption layers at the water/fluid interface. AB - The adsorption behaviour of proteins and systems mixed with surfactants of different nature is described. In the absence of surfactants the proteins mainly adsorb in a diffusion controlled manner. Due to lack of quantitative models the experimental results are discussed partly qualitatively. There are different types of interaction between proteins and surfactant molecules. These interactions lead to protein/surfactant complexes the surface activity and conformation of which are different from those of the pure protein. Complexes formed with ionic surfactants via electrostatic interaction have usually a higher surface activity, which becomes evident from the more than additive surface pressure increase. The presence of only small amounts of ionic surfactants can significantly modify the structure of adsorbed proteins. With increasing amounts of ionic surfactants, however, an opposite effect is reached as due to hydrophobic interaction and the complexes become less surface active and can be displaced from the interface due to competitive adsorption. In the presence of non-ionic surfactants the adsorption layer is mainly formed by competitive adsorption between the compounds and the only interaction is of hydrophobic nature. Such complexes are typically less surface active than the pure protein. From a certain surfactant concentration of the interface is covered almost exclusively by the non-ionic surfactant. Mixed layers of proteins and lipids formed by penetration at the water/air or by competitive adsorption at the water/chloroform interface are formed such that at a certain pressure the components start to separate. Using Brewster angle microscopy in penetration experiments of proteins into lipid monolayers this interfacial separation can be visualised. A brief comparison of the protein adsorption at the water/air and water/n-tetradecane shows that the adsorbed amount at the water/oil interface is much stronger and the change in interfacial tension much larger than at the water/air interface. Also some experimental data on the dilational elasticity of proteins at both interfaces measured by a transient relaxation technique are discussed on the basis of the derived thermodynamic model. As a fast developing field of application the use of surface tensiometry and rheometry of mixed protein/surfactant mixed layers is demonstrated as a new tool in the diagnostics of various diseases and for monitoring the progress of therapies. PMID- 10798351 TI - Effect of surfactant interfacial orientation/aggregation on adsorption dynamics. AB - The application of new thermodynamic adsorption isotherms allow to improve the description of surfactant adsorption kinetics based on a diffusional transport. While the consideration of interfacial reorientation corrects apparently too high diffusion coefficients, interfacial aggregation avoids too small diffusion coefficients or the assumption of adsorption barriers. The adsorption kinetics of alkyl dimethyl phosphine oxides is influenced by interfacial reorientation. While the lower homologues (C8-C12) follow the classical diffusion model, the higher homologues (C13-C15) yield diffusion coefficients several times larger than the physically reasonable values. Assuming two different adsorption states, the resulting diffusion coefficients agree with those expected from the geometric size of the molecules. The model also works well for oxyethylated non-ionics, such as C10EO8. As a second example, a good theoretical description is obtained for experiments of 1-decanol solutions when a mean surface aggregation number of n = 2.5 is assumed. The same n was obtained from the description of the equilibrium adsorption isotherm of 1-decanol. Assuming that the transition from one into the other state is controlled by a rate constant (change in orientation, formation or disintegration of two-dimensional aggregates) significant changes in the kinetics curves can result. The use of additional rate constants yields an improved fitting to experimental data. PMID- 10798352 TI - Penetration of dissolved amphiphiles into two-dimensional aggregating lipid monolayers. AB - The review demonstrates the recent theoretical and experimental progress in the understanding of penetration systems at the air-water interface in which a dissolved amphiphile (surfactant, protein) penetrates into a Langmuir monolayer. The critical review of the existing theoretical models which describe the thermodynamics of the penetration are critically reviewed. Although a rigorous thermodynamic analysis of penetration systems is unavailable due to their complexity, some model assumptions, e.g. the invariability of the activity coefficient of the insoluble component of the monolayer during the penetration of the soluble component results in reasonable solutions. New theoretical models describing the equilibrium behaviour of the insoluble monolayers which undergo the 2D aggregation in the monolayer, and the equations of state and adsorption isotherms which assume the existence of multiple states (conformations) of a protein molecule within the monolayer and the non-ideality of the adsorbed monolayers are now available. The theories which describe the penetration of a soluble surfactant into the main phases of Langmuir monolayers were presented first for the case of the mixture of the molecules possessing equal partial molar surfaces (the mixture of homologues), with further extension of the models to include the interesting process of the protein penetration into the monolayer of 2D aggregating phospholipid. This extension was based on a concept which subdivides the protein molecules into independent fragments with areas equal to those of the phospholipid molecule. Various mechanisms for the effect of the soluble surfactant on the aggregation of the insoluble component were considered in the theoretical models: (i) no effect on the aggregate formation process; (ii) formation of mixed aggregates; and (iii) the influence on the aggregating process via the change of aggregation constant, but without any formation of mixed aggregates. Accordingly depending on the mechanism, different forms of the equations of state of the monolayer and of the adsorption isotherms of soluble surfactant are predicted. Based on the shape of the experimental pi-A isotherms, interesting conclusions can be drawn on the real mechanism. First experimental evidence has been provided that the penetration of different proteins and surfactants into a DPPC monolayer in a fluid-like state induces a first order main phase transition of pure DPPC. The phase transition is indicated by a break point in the pi(t) penetration kinetics curves and the domain formation by BAM. Mixed aggregates of protein with phospholipid are not formed. These results agree satisfactorily with the predictions of the theoretical models. New information on phase transition and phase properties of Langmuir monolayers penetrated by soluble amphiphiles are obtained by coupling of the pi(t) penetration kinetics curves with BAM and GIXD measurements. The GIXD results on the penetration of beta-lactoglobulin into DPPC monolayers have shown that protein penetration occurs without any specific interactions with the DPPC molecules and the condensed phase consists only of DPPC. PMID- 10798353 TI - Characterization of a novel 21-kb deletion, CFTRdele2,3(21 kb), in the CFTR gene: a cystic fibrosis mutation of Slavic origin common in Central and East Europe. AB - We report a large genomic deletion of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, viz., a deletion that is frequently observed in Central and Eastern Europe. The mutation, termed CFTRdele2,3(21 kb), deletes 21,080 bp spanning introns 1-3 of the CFTR gene. Transcript analyses have revealed that this deletion results in the loss of exons 2 and 3 in epithelial CFTR mRNA, thereby producing a premature termination signal within exon 4. In order to develop a simple polymerase chain reaction assay for this allele, we defined the end-points of the deletion at the DNA sequence level. We next screened for this mutation in a representative set of European and European derived populations. Some 197 CF patients, including seven homozygotes, bearing this mutation have been identified during the course of our study. Clinical evaluation of CFTRdele2,3(21 kb) homozygotes and a comparison of compound heterozygotes for deltaF508/CFTRdele2,3(21 kb) with pairwise-matched deltaF508 homozygotes indicate that this deletion represents a severe mutation associated with pancreatic insufficiency and early age at diagnosis. Current data show that the mutation is particularly common in Czech (6.4% of all CF chromosomes), Russian (5.2%), Belorussian (3.3%), Austrian (2.6%), German (1.5%), Polish (1.5%), Slovenian (1.5%), Ukrainian (1.2%), and Slovak patients (1.1%). It has also been found in Lithuania, Latvia, Macedonia and Greece and has sporadically been observed in Canada, USA, France, Spain, Turkey, and UK, but not in CF patients from Bulgaria, Croatia, Romania or Serbia. Haplotype analysis has identified the same extragenic CF-haplotype XV-2c/KM. 19 "A" and the same infrequent intragenic microsatellite haplotype 16-33-13 (IVS8CA-IVS 17bTA-IVS 17bCA) in all examined CFTRdele2,3(21 kb) chromosomes, suggesting a common origin for this deletion. We conclude that the 21-kb deletion is a frequent and severe CF mutation in populations of Eastern- and Western-Slavic descent. PMID- 10798354 TI - SOX14 is a candidate gene for limb defects associated with BPES and Mobius syndrome. AB - Members of the SOX gene family encode proteins with homology to the HMG box DNA binding domain of SRY, the Y-linked testis-determining gene. SOX genes are expressed during embryogenesis and are involved in the development of a wide range of different tissues. Mutations in SRY, SOX9 and SOX10 have been shown to be responsible for XY sex reversal, campomelic dysplasia and Waardenburg Hirschsprung disease, respectively. It is likely that mutations in other SOX genes are responsible for a variety of human genetic diseases. SOX14 has been identified from a human genomic library and the mouse and chicken sequences obtained by polymerase chain reaction amplification. The SOX14 amino acid sequence is highly conserved across these species, suggesting an important role for this protein in vertebrate development. SOX14 is expressed in the neural tube and apical ectodermal ridge of the developing chicken limb. This is the only SOX gene known to be expressed in the apical ectodermal ridge, a structure that directs outgrowth of the embryonic limb bud. Human SOX14 is localised to a 1.15 Mb yeast artificial chromosome on chromosome 3q23, close to loci for BPES (blepharophimosis, ptosis, epicanthus inversus syndrome) and Mobius syndrome. Although SOX14 maps outside these loci, its expression pattern and chromosomal localisation suggest that it is a candidate gene for the limb defects frequently associated with these syndromes. PMID- 10798355 TI - Identification of a gene disrupted by inv(11)(q13.5;q25) in a patient with left right axis malformation. AB - An inv(11)(q13.5;q25) inversion was previously identified in a 9-month-old male patient with complex cyanotic heart defects, altered lung lobation, symmetric liver, and abnormally lobulated spleen (polysplenia). This chromosomal rearrangement was inherited from the phenotypically normal father. We termed these regions DHTX-A (disrupted in heterotaxy)-- A at 11q13.5 and DHTX-B at 11q25. Here, we report the isolation and characterization of the inversion breakpoints and the gene that is disrupted by the DHTX-A breakpoint. The putative DHTX is identical to the UVRAG gene, which was originally identified as a gene that complements the UV sensitivity of xeroderma pigmentosum complementation group C. The 4-kb mRNA was found to be encoded by a large gene, at least 300 kb long, composed of 15 exons. The function of the gene product remains largely unknown. However, the near central portion of the UVRAG protein is predicted to contain a coiled-coil domain, which has been implicated in mediating protein protein interactions. Southern analyses and fluorescence in situ hybridization (FISH) revealed that the DHTX-A breakpoint in the patient and his father lies within the intron between exons 6 and 7 of UVRAG. Northern blot analysis indicated strong expression in human fetal and adult tissues and in mouse embryonic day-7 and adult tissues, respectively. Whole mount in situ hybridization also showed that the Uvrag gene is expressed in the presomite-stage embryo. Several hypotheses are discussed to explain the relationship between the chromosomal inversion and the accompanying phenotypes. PMID- 10798356 TI - Identification of 187 single nucleotide polymorphisms (SNPs) among 41 candidate genes for ischemic heart disease in the Japanese population. AB - To investigate whether common variants in the human genetic background are associated with pathogenesis of ischemic heart diseases, we systematically surveyed 41 possible candidate genes for single-nucleotide polymorphisms (SNPs) by directly sequencing 96 independent alleles at each locus, derived from 48 unrelated Japanese patients with myocardial infarction, including 25.8 kb 5' flanking regions, 56.8 kb exonic and 35.4 kb intronic sequences, and 1.8 kb 3' flanking regions. In this genomic DNA of nearly 120 kb, we identified 187 SNPs: 55 in 5' flanking regions, seven in 5' untranslated regions (UTRs), 52 in coding elements, 64 in introns, eight in 3' UTRs, and one in a 3' flanking region. Among the 52 coding SNPs, 26 were non-synonymous changes. Allelic frequencies of some of the polymorphisms were significantly different from those reported in European populations. For example, the Q506R substitution in the coagulation factor V gene, the so-called "Leiden mutation", has a reported frequency of 2.3% in Europeans, but we detected the Leiden mutation in none of the Japanese genomes that we investigated. The allelic frequencies of the -33A>G SNP in the thrombomodulin gene were also very different; this allele occurred at a 12% frequency in the Japanese patients that we examined, although it had been detected in none of 82 Caucasians reported previously. These data support the hypothesis that some SNPs are specific to particular ethnic groups. PMID- 10798357 TI - Identification of 142 single nucleotide polymorphisms in 41 candidate genes for rheumatoid arthritis in the Japanese population. AB - Single-nucleotide polymorphisms (SNPs) can make an important contribution to our understanding of genetic backgrounds that may influence medical conditions and ethnic diversity. We undertook a systematic survey of genomic DNA for SNPs located not only in coding sequences but also in non-coding regions (e.g., introns and 5' flanking regions) of selected genes. Using DNA samples from 48 Japanese patients with rheumatoid arthritis (RA) as templates, we surveyed 41 genes that represent candidates for RA, screening a total of 104 kb of DNA (30 kb of coding sequences and 74 kb of non-coding DNA). Within this 104 kb of genomic sequences we identified 163 polymorphisms (1 per 638 bases on average), of which 142 were single-nucleotide substitutions and the remainder, insertions or deletions. Of the coding SNPs, 52% were non-synonymous substitutions, and non conservative amino acid changes were observed in a quarter of those. Sixty-nine polymorphisms showed high frequencies for minor alleles (more than 15%) and 20 revealed low frequencies (<5%). Our results indicated a greater average distance between SNPs than others have reported, but this disparity may reflect the type of genes surveyed and/or the relative ethnic homogeneity of our test population. PMID- 10798358 TI - Human glycine decarboxylase gene (GLDC) and its highly conserved processed pseudogene (psiGLDC): their structure and expression, and the identification of a large deletion in a family with nonketotic hyperglycinemia. AB - Mutations in the glycine decarboxylase gene (GLDC) cause nonketotic hyperglycinemia (NKH), an in-born error of metabolism characterized by severe neurological disturbance. We have determined the structure of GLDC and of its pseudogene (psiGLDC) and studied their expression for a molecular analysis of NKH. The GLDC gene spans at least 135 kb and consists of 25 exons. All donor and acceptor sites adhere to the canonical GT-AG rule, except for the donor site of intron 21, where a variant form GC is used instead of GT. The transcription initiation site has been assigned to a residue 163 bp upstream from the translation initiation triplet by primer extension analysis. The psiGLDC gene has no intron and shares 97.5% homology with the coding region of functional GLDC, suggesting that psiGLDC is a processed pseudogene that arose from the GLDC transcript about 4-8 million years ago. RNA blotting analysis has revealed that GLDC is expressed in human liver, kidney, brain, and placenta. We have also examined a patient with NKH with no detectable GLDC mRNA in his lymphoblasts. Exons 1-3 of the functional GLDC gene from this patient are not amplified by polymerase chain reaction (PCR), whereas those from control subjects are. These results suggest a large homozygous deletion (at least 30 kb) in the patient. Furthermore, we have devised a semi-quantitative PCR to estimate the number of GLDC alleles by using psiGLDC as an internal control and have confirmed the homozygosity and heterozygosity of the deletion in the patient and his parents, respectively. Structural information of GLDC and psiGLDC should facilitate the molecular analysis of NKH. PMID- 10798359 TI - Del(X)(p21.1) in a mother and two daughters: genotype-phenotype correlation of Turner features. AB - We report a mother and two daughters with partial Xp monosomy. Clinical assessment for Turner phenotype revealed that the three females manifested low normal to mild short stature (-1.6 to approximately -2.3 SD) and variable degrees of skeletal features, such as cubitus valgus, short 4th matacarpals, and Madelung deformity, but no soft tissue or visceral anomalies or gonadal dysfunction. Cytogenetic studies for lymphocytes showed that the karyotype was 45,X[3]/46,X,del(X)(p21.1)[27] in the mother and non-mosaic 46,X,del(X)(p21.1) in the two daughters. Fluorescence in situ hybridization and microsatellite analyses for 19 loci/regions on the X chromosome demonstrated that the del(Xp) chromosome was missing SHOX and had the breakpoint between DMD and CYBB. The results are consistent with the recently proposed notion that haploinsufficiency of SHOX results in not only short stature, but also Turner skeletal features in association with maturational effects of gonadal estrogens. The lack of soft tissue or visceral anomalies suggests the presence of the putative lymphogenic gene on the del(Xp) chromosome; the preservation of ovarian function appears to be compatible with meiotic pairing failure being relatively mild. PMID- 10798360 TI - Illegitimate splicing of the NF1 gene in healthy individuals mimics mutation induced splicing alterations in NF1 patients. AB - Neurofibromatosis type 1 (NF1) is a common inherited disease affecting one in 3,500 individuals. The mutation rate in the NF1 gene is one of the highest known for human genes. Compared to other methods, the protein truncation test (PTT) and subsequent sequence analysis of cloned cDNA provides improved efficiency in detecting NF1 mutations that are dispersed throughout the gene spanning 350 kb of genomic DNA. Sequencing of cDNA of patients affected with NF1 mutations revealed multiple splicing errors. Since similar missplicings were also found in "aged" blood of healthy individuals, they are most likely attributable to a general decrease in splice site selection in aged blood. We show that restoring viability of lymphocytes before RNA extraction by cultivation and PHA stimulation diminishes aberrant splicing in aged blood and is thus useful to circumvent splicing alterations which are frequently compromising mutation detection in patient samples and mimic mutation-induced alterations of mRNA. PMID- 10798361 TI - Association between M467T and 114 C-->A variants within the SLC3A1 gene and some phenotypical traits in cystinuria patients from Spain. AB - Cystinuria is an inherited metabolic disease characterized by an abnormal urinary excretion of cystine and dibasic amino acids. Formation of renal calculi, recurrent infections and renal failure are the main complications of this disease. The SLC3A1 gene, which codes for a dibasic amino acid transporter protein, is involved in the pathogenesis of cystinuria. We investigated the possible association between molecular variants (M467T, E483X, T216 M and 114 C- >A) within the SLC3A1 gene and some phenotypical traits in a Spanish area. The study population consisted of 45 cystinuria patients, 42 cystinuria relatives and 81 healthy control subjects. Only the M467T mutation was found in chromosomes of cystinuria patients and relatives. However, the 114 C-->A polymorphism was detected in cystinuria patients, in relatives and in control subjects but with different prevalences. Moreover, a statistically significant association between this polymorphism and urinary amino acid levels was found in cystinuria patients (P<0.05). Subjects with the C/C genotype showed significantly higher urinary levels of cystine, arginine and their sum as compared with carriers of the A allele (P<0.05). When multiple linear regression analysis was performed in cystinuria patients, the 114 C-->A polymorphism remained significantly associated (P=0.047) with cystine levels even after controlling for age, gender and the M467T mutation. Furthermore, we also found a statistically significant interaction term (P=0.028) between M467T and 114 C-->A in determining urinary cystine levels. According to our results, the 114 C-->A polymorphism might be a marker of a functional variant in the SLC3A1 gene or in other genes related to urinary amino acid excretion in cystinuria patients. PMID- 10798362 TI - The spectrum of mutations in erythrokeratodermias--novel and de novo mutations in GJB3. AB - Intercellular channels in skin are a complex and functionally diverse system formed by at least eight connexins (Cx). Our recent molecular studies implicating Cx defects in inherited skin disorders emphasize the critical role of this signaling pathway in epidermal differentiation. Erythrokeratodermia variabilis (EKV) is an autosomal dominant genodermatosis with a striking phenotype characterized by the independent occurrence of transient localized erythema and hyperkeratosis. The disease maps to 1p34-p35, and recently we identified the causative gene GJB3 encoding Cx31. We have now investigated GJB3 in two families and three sporadic cases with EKV, and report three new heterozygous mutations. In a sporadic case, we detected a mutation leading to substitution of a conserved phenylalanine (F137L) in the third transmembrane domain, which likely interferes with the proper assembly or gating properties of connexons. In another family, all three affected individuals carried two distinct mutations on the same GJB3 allele. However, only a de novo heterozygous missense mutation replacing arginine 42 with proline (R42P) co-segregated with the disease, while a 12 bp deletion predicted to eliminate four amino acid residues in the variable carboxy terminal domain of Cx31 was also found in clinically unaffected relatives but not in 90 unaffected controls. Including the previously published mutations, in toto, five different missense mutations have now been detected in 6 out of 17 families investigated by our laboratory, all of which presumably affect the cytoplasmic amino terminal and transmembrane domains of Cx31. In contrast, two mutations linked to progressive high-tone hearing impairment were located in the second extracellular domain, suggesting that the character and position of Cx mutations determine their phenotypic expression in different tissues. However, the phenotypic spectrum of GJB3 mutations seems not to include progressive symmetric erythrokeratodermia, another dominant genodermatosis with overlapping features, since no mutations were found in six unrelated families tested. PMID- 10798363 TI - A 3D model of human P450c21: study of the putative effects of steroid 21 hydroxylase gene mutations. AB - In order to better understand the disease-causing role of missense mutations found in the CYP21 gene from patients affected with congenital adrenal hyperplasia (CAH) due to steroid 21-hydroxylase deficiency, we built two three dimensional (3D) models of human P450c21 using all known 3D structures of P450s. For each residue affected by a missense mutation, its location in the 3D structure and the putative changes in terms of biochemical properties brought about by the mutation were analyzed. Most of the severe alleles were found to affect residues located in functionally important regions of the molecule such as substrate recognition sites (SRS) or the heme region, whereas moderate mutations were mostly found in less crucial regions of the molecule. Thus, there is a good correlation between the 3D structure study and clinical data and mutagenesis experiments previously reported. In one case, however, the observed clinical severity of the mutation (E380D) did not match its expected severity deduced from the model, pointing to a potential functionally important region of the molecule. Our 3D human models provide a basic model for further studies of mutations responsible for 21-hydroxylase, and for identification of important residues involved in the specific activity of the enzyme. PMID- 10798364 TI - Parental origin of the extra chromosome in prenatally diagnosed fetal trisomy 21. AB - Trisomy 21 (Down syndrome) is one of the most common chromosomal abnormalities. Of cases of free trisomy 21 causing Down syndrome, about 95% result from nondisjunction during meiosis, and about 5% are due to mitotic errors in somatic cells. Previous studies using DNA polymorphisms of chromosome 21 showed that paternal origin of trisomy 21 occurred in only 6.7% of cases. However, these studies were conducted in liveborn trisomy 21-affected infants, and the possible impact of fetal death was not taken into account. Using nine distinct DNA polymorphisms, we tested 110 families with a prenatally diagnosed trisomy 21 fetus. Of the 102 informative cases, parental origin was maternal in 91 cases (89.2%) and paternal in 11 (10.8%). This percentage differs significantly from the 7.0% observed in previous studies (P<0.001). In order to test the influence of genomic parental imprinting, we determined the origin of the extra chromosome 21 in relation to different factors: advanced maternal age, maternal serum human chorionic gonadotropin (hormone of placental origin), severity of the disease, gestational age at diagnosis and fetal gender. We found that the increased frequency of paternal origin of nondisjunction in trisomy 21-affected fetuses cannot obviously be explained by factors leading to selective loss of paternal origin fetuses. PMID- 10798365 TI - Identification of susceptibility loci for nonsyndromic cleft lip with or without cleft palate in a two stage genome scan of affected sib-pairs. AB - Nonsyndromic cleft lip with or without cleft palate (CL/P) is a complex disorder of multigenic origin involving between two and ten loci. Linkage and association studies of CL/P have implicated a number of candidate genes and regions but have often proved difficult to replicate. Here, we report the findings from a two stage genome-wide scan of 92 affected sib-pairs to identify susceptibility loci to CL/P. An initial set of 400 microsatellite markers was used, with an average spacing of 10 cM throughout the genome. Eleven regions on eight chromosomes were found to have a P-value smaller than 0.05. These eight chromosomes were then further mapped with a second set of markers to increase the average map density to 5 cM. In seven out of eleven areas densely mapped, significance was markedly increased by decreasing the marker interval. Excessive allele sharing was found at 1p (NPL=2.35, P=0.009, MLS=1.51), 2p (NPL=1.77, P= 0.04, MLS=0.66), 6p (NPL=2.35, P=0.009, MLS=1.34), 8q (NPL=2.15, P=0.015, MLS= 1.51) 11 cen (NPL=2.70, P=0.003, MLS=2.10), 12q (NPL=2.08, P=0.02, MLS= 1.5), 16p (NPL=2.1, P=0.018, MLS=0.97) and Xcen-q (NPL=2.40, P=0.008, MLS=2.68). Although none reached the level required for significant susceptibility loci, two of these areas have previously been implicated in CL/P, viz. 2p13, an area harbouring the TGFA gene, and 6p23-24. We also demonstrate highly suggestive linkage to a susceptibility locus for nonsyndromic clefting on the X chromosome. Further studies are currently underway to replicate these findings in a larger cohort of affected sib-pairs. PMID- 10798366 TI - Linkage of recessive Robinow syndrome to a 4 cM interval on chromosome 9q22. AB - Autosomal recessive Robinow syndrome is a form of mesomelic dwarfism with multiple rib and vertebral anomalies. Using autozygosity mapping we have identified a genetic locus (RBNW1) for this syndrome at chromosome 9q22 in seven consanguineous families from Oman. Our results indicate that the gene lies within a 4 cM region between markers D9S1836 and D9S1803 (maximum multipoint LOD score 12.3). In addition, we have analysed two non-Omani families with autosomal recessive Robinow and found no genetic heterogeneity. PMID- 10798367 TI - Segregation analysis of serum uric acid in the NHLBI Family Heart Study. AB - Segregation analysis was performed on the serum uric acid measurements from 523 randomly ascertained Caucasian families from the NHLBI Family Heart Study. Gender specific standardized residuals were used as the phenotypic variable in both familial correlation and segregation analysis. Uric acid residuals were adjusted for age, age2, age3, body mass index (kg/m2), creatinine level, aspirin use (yes/no), total drinks (per week), HOMA insulin resistance index [(glucose * insulin)/22.5], diuretic use (yes/no), and triglyceride level. Sibling correlations (r=0.193) and parent-offspring correlations (r=0.217) were significantly different from zero, but these two familial correlations were not significantly different from one another. After adjustment for covariates, the heritability estimate for serum uric acid was 0.399. Segregation analysis rejected the "no major gene" model but was unable to discriminate between an "environmental" and a "Mendelian major gene" model. These results support the hypothesis that uric acid is a multifactorial trait possibly influenced by more than one major gene, modifying genes, and environmental factors. PMID- 10798368 TI - Spectrum of CFTR mutations in Mexican cystic fibrosis patients: identification of five novel mutations (W1098C, 846delT, P750L, 4160insGGGG and 297-1G-->A). AB - We have analyzed 97 CF unrelated Mexican families for mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Our initial screening for 12 selected CFTR mutations led to mutation detection in 56.66% of the tested chromosomes. In patients with at least one unknown mutation after preliminary screening, an extensive analysis of the CFTR gene by single stranded conformation polymorphism (SSCP) or by multiplex heteroduplex (mHET) analysis was performed. A total of 34 different mutations representing 74.58% of the CF chromosomes were identified, including five novel CFTR mutations: W1098C, P750L, 846delT, 4160insGGGG and 297-1G-->A. The level of detection of the CF mutations in Mexico is still lower than that observed in other populations with a relatively low frequency of the deltaF508 mutation, mainly from southern Europe. The CFTR gene analysis described here clearly demonstrated the high heterogeneity of our CF population, which could be explained by the complex ethnic composition of the Mexican population, in particular by the strong impact of the genetic pool from southern European countries. PMID- 10798369 TI - A fragile X case with an amplification/deletion mosaic pattern. AB - Fragile X syndrome is the most common cause of hereditary mental retardation. The FMR1 gene, which is involved in fragile X syndrome, contains a polymorphic CGG repeat, which expands in affected patients. Expanding triplet repeats have been shown to be a new type of mutation, termed "dynamic mutation", responsible for more than 12 genetic diseases. These mutations occur as multiple steps rather than as a single event. The first step leads to an unstable allele that then becomes increasingly unstable generally achieving further increases in copy or occasionally contraction. In this report, we describe a fragile X boy with both a hypermethylated full mutation and a deletion of 905 bp encompassing the CGG repeat. The upstream breakpoint is 438 bp 5' to the CGG repeat and the downstream breakpoint is 420 bp 3' of the triplet repeats. The deletion includes the ATG starting codon for translation of the FMR1 gene. This was confirmed by using FMRP immunocytochemistry both on blood smears and hair roots. The deleted region is flanked by a ccgg direct repeat next to the breakpoints; this may have had a critical role in the formation of a secondary DNA structure leading to the deletion. PMID- 10798370 TI - Characterization of a partial pseudogene homologous to the Hermansky-Pudlak syndrome gene HPS-1; relevance for mutation detection. AB - The HPS-1 gene is the first gene found to be responsible for the autosomal recessive disorder Hermansky-Pudlak syndrome (HPS). HPS is characterized by oculocutaneous albinism, a platelet storage pool deficiency, and ceroid lipofuscinosis. The HPS-1 gene has been mapped to chromosome 10q23.1-23.3 and encodes a 79-kDa protein of unknown function with no homology to any known protein. A sequence database search has revealed that a portion of clone HS 1119A7 shows high sequence similarity to HPS-1 cDNA. By performing sequence alignments and PCR amplification of cDNA from several human tissues, we have shown that part of this clone consists of an unprocessed partial HPS-1 pseudogene located on chromosome 22q12.2-12.3. The pseudogene contains several intact HPS-1 exons and shows 95% sequence homology to the HPS-1 cDNA. Exon 6 of the pseudogene has 100% sequence homology to exon 6 of HPS-1 itself. In the pseudogene, this exon is surrounded by portions of both its normal flanking introns. These data provide the first characterization of an HPS-1 pseudogene, called HPS1-psi1. During amplification of exon 6 of the HPS-1 gDNA for mutation identification, the pseudogene might also be amplified, leading to a false positive for mutation. In addition, amplification of specific parts of the HPS-1 cDNA (e.g., exons 2-5) for mutation detection might lead to false positives for mutations, if the cDNA is contaminated with gDNA. This calls for caution when employing these screening approaches. PMID- 10798371 TI - Context-dependent conformation of diethylglycine residues in peptides. AB - Diethylglycine (Deg) residues incorporated into peptides can stabilize fully extended (C5) or helical conformations. The conformations of three tetrapeptides Boc-Xxx-Deg-Xxx-Deg-OMe (Xxx=Gly, GD4; Leu, LD4 and Pro, PD4) have been investigated by NMR. In the Gly and Leu peptides, NOE data suggest that the local conformations at the Deg residues are fully extended. Low temperature coefficients for the Deg(2) and Deg(4) NH groups are consistent with their inaccessibility to solvent, in a C5 conformation. NMR evidence supports a folded beta-turn conformation involving Deg(2)-Gly(3), stabilized by a 4-->1 intramolecular hydrogen bond between Pro(1) CO and Deg(4) NH in the proline containing peptide (PD4). The crystal structure of GD4 reveals a hydrated multiple turn conformation with Gly(1)-Deg(2) adopting a distorted type II/II' conformation, while the Deg(2)-Pro(3) segment adopts a type III/III' structure. A lone water molecule is inserted into the potential 4-->1 hydrogen bond of the Gly(1)-Deg(2) beta-turn. PMID- 10798372 TI - Solvolysis and aminolysis on peptidyl-Kaiser oxime resin assisted by Ca2+ and Eu3+: a mild procedure to prepare alpha-methyl and -ethyl esters of protected peptides. AB - Ca2+ and Eu3+ were able to assist solvolysis on peptidyl-Kaiser oxime resins generating alpha-methyl and -ethyl esters of protected peptides. The methanolysis assistance was at least twice as effective as that of acetic acid, the common catalyst used in aminolysis of the ester oxime linkage. No molar excess of Ca2+ or Eu3+ was needed to enhance this reaction efficiency. Ca2+ also assisted aminolysis on peptidyl-Kaiser oxime resins. Solvolysis and aminolysis rates depended on the nature of the C-terminal residue attached to the resin and on the alcohol used. Both reactions were selective to the ester oxime linkage since no significant amount of secondary products, resulting from rearrangements or simultaneous transesterification of the beta-benzyl or cyclohexyl esters, was detected in the reaction media. The alpha-methyl and -ethyl esters of Ac-Ala-Gly X [where, X = Gly, Ala, Phe or Lys (2-Cl-Z)] and of Ac-Ile-Ser (Bzl)-Asp(OZ) (where, Z = Bzl or cHex) were essentially the only products formed in the solvolyses performed. Ac-Ile-Ser(Bzl)-Asp(OcHex)Arg(HCl)-OMe and Ac-Ile-Ser(Bzl) Asp(OcHex)Arg (HCl)-OEt were the major products formed in the aminolysis reactions. In the presence of the metal ions, the resin-cleavage yields were > 50%. In their absence, they were < 15%. PMID- 10798373 TI - Convenient and efficient synthesis of Boc-/Z-/Fmoc-beta-amino acids employing N protected alpha-amino acid fluorides. AB - A new and efficient method for the synthesis of N(alpha)-Fmoc-/Boc-/Z-beta-amino acids using the two-step Arndt-Eistert approach is described. Fmoc-/Boc-/Z-alpha Amino acid fluorides were used for the acylation of diazomethane synthesizing Fmoc-/Boc-/Z-alpha-aminodiazoketones as crystalline solids with good yield and purity. They were then converted to the corresponding beta-amino acids using PhCOOAg/dioxane/H2O. PMID- 10798374 TI - Zinc-promoted simple synthesis of oligomer-free N(alpha)-Fmoc-amino acids using Fmoc-Cl as an acylating agent under neutral conditions. AB - A range of N(alpha)-Fmoc-protected amino acids, including those that contain t butyl moiety, have been synthesized by employing Fmoc-Cl utilizing the activated, commercial zinc dust-promoted synthesis of carbamates under neutral conditions. A general procedure is described that circumvents the oligomerization side reaction normally noticed in Schotten-Baumann conditions. It is a simple, convenient and clean method. Thus, Fmoc-amino acids are obtained in high yield (85-92%) and purity as checked by thin-layer chromatography, high-performance liquid chromatography and other physical methods. PMID- 10798375 TI - Diastereoselective synthesis of non-proteinogenic alpha-amino acids. AB - The synthesis of enantiomerically pure azatyrosine, tribromophenylalanine and trichlorophenylalanine is described, using two methods, diastereoselective alkylation and (or) diastereoselective protonation of chiral enolates. PMID- 10798376 TI - Structural features of the Pip/AzPip couple in the crystalline state: influence of the relative AzPip location in an azadipeptide sequence upon the induced chirality and conformational characteristics. AB - Azapipecolic (AzPip) is a pipecolic (Pip) residue analogue containing a nitrogen atom in place of the C(alpha)H group. AzPip was introduced into two reverse dipeptide sequences, Piv-AzPip-L-Ala-NHiPr I and Boc-L-Ala-AzPip-NHiPr II in order to evaluate, in the crystalline state, the influence of the L-Ala-induced chirality upon the prochiral AzPip residue, and therefore the resulting conformational characteristics, according to the relative position of the AzPip residue. Piv-DL-Pip-NHMe III served as a control derivative for comparison between the properties of the two different heterocycles of Pip and AzPip residues. Piperidine and hexahydropyridazine rings have a few characteristics in common: chair conformation, axial disposition of the C-terminal backbone substituent and the cisoid form of the N-terminal tertiary amide function. An almost pure sp3 hybridization state is observed for the substituted nitrogen atom N(alpha), so that L-Ala induces an AzPip (R) or (S) chirality when it follows or precedes, respectively, the azaresidue in such a pseudodipeptide sequence. If both I and II compounds present a short NH...N contact between the sp2 tertiary amide nitrogen atom and the NH of the next secondary amide function, whatever the chiral nature of the sequence, the heterochiral azadipeptide I adopts a rather totally extended conformation while the homochiral azadipeptide II is folded by a beta-VI turn-like structure stabilized by a classical 4-->1 intramolecular hydrogen bond. PMID- 10798377 TI - Isolation of leptin-binding peptides from a random peptide phage library. AB - Leptin plays a role in regulating the body weight in mice. Injection of recombinant mouse leptin expressed in Escherichia coli reduced the food intake and body weight in normal, ob/ob and diet-induced obesity mice. Hyperglycemia, hyperinsulinemia and hypothermia can also be corrected in ob/ob mice after leptin injection. Leptin is a 16-kDa secretory protein comprising 167 amino acids produced in adipose tissue and is secreted to blood stream. In this study, a recombinant mouse leptin was generated and purified from a baculovirus expression system. This protein was used to identify putative ligands using a phage library of random peptides. Three leptin-binding phage clones were found, which were characterized by DNA sequencing and ELISA methods. The amino acid sequences of the reactive peptides are: LAYCSDPVRCLVWWY, MFWISAVSFVDHALV and LVLVLSAFLCCGVG. All three clones bound to recombinant human and mouse leptins. These peptides may be useful tools to study leptin-receptor interaction, food intake and body weight regulation. PMID- 10798378 TI - Protease-catalyzed fragment condensation via substrate mimetic strategy: a useful combination of solid-phase peptide synthesis with enzymatic methods. AB - The concept of substrate mimetic strategy represents a new powerful method in the field of enzymatic peptide synthesis. This strategy takes advantage of the shift in the site-specific amino acid moiety from the acyl residue to the ester-leaving group of the carboxyl component enabling acylation of the enzyme by nonspecific acyl residues. As a result, peptide bond formation occurs independently of the primary specificity of proteases. Moreover, because of the coupling of nonspecific acyl residues, the newly formed peptide bond is not subject to secondary hydrolysis achieving irreversible peptide synthesis. Here, we report the combination of solid-phase peptide synthesis with substrate mimetic-mediated enzymatic peptide fragment condensations. First, the utility of the oxime resin strategy for the synthesis of peptide fragments in the form of substrate mimetics esterified as 4-guanidinophenyl-, phenyl- and mercaptopropionic acid esters was investigated. The study was completed by using the resulting N(alpha)-protected peptide esters as acyl donors in trypsin-, alpha-chymotrypsin- and V8 protease catalyzed fragment condensations. PMID- 10798379 TI - Conformational mapping of the N-terminal segment of surfactant protein B in lipid using 13C-enhanced Fourier transform infrared spectroscopy. AB - Synthetic peptides based on the N-terminal domain of human surfactant protein B (SP-B1-25; 25 amino acid residues; NH2-FPIPLPYCWLCRALIKRIQAMIPKG) retain important lung activities of the full-length, 79-residue protein. Here, we used physical techniques to examine the secondary conformation of SP-B1-25 in aqueous, lipid and structure-promoting environments. Circular dichroism and conventional, 12C-Fourier transform infrared (FTIR) spectroscopy each indicated a predominate alpha-helical conformation for SP-B1-25 in phosphate-buffered saline, liposomes of 1-palmitoyl-2-oleoyl phosphatidylglycerol and the structure-promoting solvent hexafluoroisopropanol; FTIR spectra also showed significant beta- and random conformations for peptide in these three environments. In further experiments designed to map secondary structure to specific residues, isotope-enhanced FTIR spectroscopy was performed with 1-palmitoyl-2-oleoyl phosphatidylglycerol liposomes and a suite of SP-B1-25 peptides labeled with 13C-carbonyl groups at either single or multiple sites. Combining these 13C-enhanced FTIR results with energy minimizations and molecular simulations indicated the following model for SP-B1-25 in 1-palmitoyl-2-oleoyl phosphatidylglycerol: beta-sheet (residues 1-6), alpha-helix (residues 8-22) and random (residues 23-25) conformations. Analogous structural motifs are observed in the corresponding homologous N-terminal regions of several proteins that also share the 'saposin-like' (i.e. 5-helix bundle) folding pattern of full-length, human SP-B. In future studies, 13C-enhanced FTIR spectroscopy and energy minimizations may be of general use in defining backbone conformations at amino acid resolution, particularly for peptides or proteins in membrane environments. PMID- 10798381 TI - TOXNET: A computerized collection of toxicological and environmental health information. AB - The Toxicology and Environmental Health Information Program, managed by the National Library of Medicine's Division of Specialized Information Services, provides access to a number of online bibliographic and factual computer files concerned with the toxicology, safety and handling, and environmental fate of chemicals, along with other files that cover genetic toxicology, developmental and reproductive toxicology, mutagenesis, carcinogenesis and toxic chemical releases. PMID- 10798380 TI - Design rationale, synthesis, and characterization of non-natural analogs of the cationic amino acids arginine and lysine. AB - A series of non-natural isosteric analogs of the cationic, ion-pairing, natural amino acids arginine and lysine have been synthesized, characterized with regard to relevant physical parameters, and protected for routine inclusion in Merrifield solid-phase synthesis. The design of these molecules is based on the concept of steric inhibition of solvation, in that judicious placement of alkyl groups can destabilize aqueous ion solvation and favor ion-pairing [see Beeson & Dix (1993) J. Am. Chem. Soc. 115, 10275]. When the residues are substituted for the natural amino acids in biologically active peptides, enhanced ion-pairing of the peptides to their receptors to increase the peptides' biological activities can result. The increased lipophilicity of the non-natural residues can also improve pharmacokinetic parameters and agonist/antagonist behaviors of peptides. While the synthesis of the L-series is described, the D-isomers were also prepared using identical chemistry. PMID- 10798382 TI - Histochemical demonstration of changes in the activity of hepatic phosphatases induced by experimental lead poisoning in male white rats (Rattus norvegicus). AB - Seven hepatic phosphatases were histochemically investigated in male white rats (Rattus norvegicus) pretreated with chronic subtoxic doses of lead acetate. Lead has increased the activities of alkaline-, acid-, neutral-, adenosine mono- and glucose-6-phosphatase, but has markedly decreased the activity of membrane-bound Na+-K+, ATPase while the activity of mitochondrial Mg2+-ATPase was not altered. It has also produced heterogenous alterations in the distribution patterns, sites of the enzymatic activities and in the intensity of phosphatase activities among the same type of cells in the terminal afferent and efferent venules of the hepatic lobules. The obtained histochemical findings indicate that the alterations in the activities of hepatic phosphatases could be an adaptation to the metabolic, structural and functional changes in the organelles of hepatic cells due to lead intoxication. PMID- 10798383 TI - Correlations among tumor types in mouse cancer bioassays: liver adenomas, liver carcinomas, leukemias and lymphomas. AB - In an examination of rodent bioassays Young and Gries [Young S.S., and Gries C.L. Exploration of the negative correlation between proliferative hepatocellular lesions and lymphoma in rats and mice--establishment and implications. Fundam. Appl. Toxicol. 1984: 4: 632-6401 and Haseman et al. [Haseman J.K., et al. Body weight-tumor incidence correlations in long-term rodent carcinogenicity studies. Toxicol. Pathol. 1997: 25: 256-263] noticed that there is a negative correlation (anticorrelation) between development of liver tumors and leukemia or lymphomas. If an animal has a lymphoma or leukemia it is less likely to develop liver tumors. These studies noted that this applies to several strains of animals. The anticorrelation appeared in control animals. In this paper we study this anticorrelation in the quarter of a million rodents exposed in the Carcinogenesis Bioassay Database System (CBDS) database of the National Toxicology program in both control and dosed animals. We failed to completely replicate Young and Gries or Haseman et al. However, when benign liver tumors (adenomas or nodules) and malignant liver tumors (carcinomas) are considered separately AND different leukemia and lymphoma types are considered, a strong anticorrelation appears. We identify survival and the time period (in years) during which the bioassay was completed as an important factor in interpreting correlations and anticorrelations. Differences between liver adenomas and carcinomas and lymphoma types contribute to a more general question of grouping of the individual tumor types. In our classification scheme (originally developed about 1986 by Dr. Bailar) an effort is made to distinguish benign and malignant neoplasms, while other investigators group all tumors at a specific site. For many analyses liver adenomas and carcinomas have been lumped together. This is because it is suspected that the diagnoses by pathologists may not distinguish, or ignore the (benign) adenoma when a (malignant) carcinoma is present. Possible biological differences in tumor mechanisms may require separate evaluation of these tumors with all the dangers of "pathologist bias" that this introduces. PMID- 10798384 TI - Trio quartet in D. (melanogaster) PMID- 10798385 TI - Integrating with action potentials. PMID- 10798386 TI - AMPA receptors unbound: membrane cycling and synaptic plasticity. PMID- 10798387 TI - Polyglutamine expansions: proteolysis, chaperones, and the dangers of promiscuity. PMID- 10798388 TI - From ionic currents to molecular mechanisms: the structure and function of voltage-gated sodium channels. PMID- 10798389 TI - Addiction becomes a brain disease. PMID- 10798390 TI - Ion channels and synaptic organization: analysis of the Drosophila genome. PMID- 10798391 TI - A genome-wide search for synaptic vesicle cycle proteins in Drosophila. AB - The recent completion of the Drosophila genome sequence opens new avenues for neurobiology research. We screened the fly genome sequence for homologs of mammalian genes implicated directly or indirectly in exocytosis and endocytosis of synaptic vesicles. We identified fly homologs for 93% of the vertebrate genes that were screened. These are on average 60% identical and 74% similar to their vertebrate counterparts. This high degree of conservation suggests that little protein diversification has been tolerated in the evolution of synaptic transmission. Finally, and perhaps most exciting for Drosophila neurobiologists, the genomic sequence allows us to identify P element transposon insertions in or near genes, thereby allowing rapid isolation of mutations in genes of interest. Analysis of the phenotypes of these mutants should accelerate our understanding of the role of numerous proteins implicated in synaptic transmission. PMID- 10798392 TI - Dynamic statistical parametric mapping: combining fMRI and MEG for high resolution imaging of cortical activity. AB - Functional magnetic resonance imaging (fMRI) can provide maps of brain activation with millimeter spatial resolution but is limited in its temporal resolution to the order of seconds. Here, we describe a technique that combines structural and functional MRI with magnetoencephalography (MEG) to obtain spatiotemporal maps of human brain activity with millisecond temporal resolution. This new technique was used to obtain dynamic statistical parametric maps of cortical activity during semantic processing of visually presented words. An initial wave of activity was found to spread rapidly from occipital visual cortex to temporal, parietal, and frontal areas within 185 ms, with a high degree of temporal overlap between different areas. Repetition effects were observed in many of the same areas following this initial wave of activation, providing evidence for the involvement of feedback mechanisms in repetition priming. PMID- 10798393 TI - Formation of precise connections in the olfactory bulb occurs in the absence of odorant-evoked neuronal activity. AB - Olfactory neurons expressing the same odorant receptor converge to a small number of glomeruli in the olfactory bulb. In turn, mitral and tufted cells receive and relay this information to higher cortical regions. In other sensory systems, correlated neuronal activity is thought to refine synaptic connections during development. We asked whether the pattern of connections between olfactory sensory axons and mitral cell dendrites is affected when odor-evoked signaling is eliminated in mice lacking functional olfactory cyclic nucleotide-gated (CNG) channels. We demonstrate that olfactory sensory axons converge normally in the CNG channel mutant background. We further show that the pruning of mitral cell dendrites, although slowed during development, is ultimately unperturbed in mutant animals. Thus, the olfactory CNG channel-and by inference correlated neural activity--is not required for generating synaptic specificity in the olfactory bulb. PMID- 10798394 TI - Peripheral olfactory projections are differentially affected in mice deficient in a cyclic nucleotide-gated channel subunit. AB - Axons of olfactory sensory neurons expressing a given odorant receptor converge to a few glomeruli in the olfactory bulb. We have generated mice with unresponsive olfactory sensory neurons by targeted mutagenesis of a cyclic nucleotide-gated channel subunit gene, OCNC1. When these anosmic mice were crossed with mice in which neurons expressing a given odorant receptor can be visualized by coexpression of an axonal marker, the pattern of convergence was affected for one but not another receptor. In a novel paradigm, termed monoallelic deprivation, axons from channel positive or negative neurons that express the same odorant receptor segregate into distinct glomeruli within the same bulb. Thus, the peripheral olfactory projections are in part influenced by mechanisms that depend on neuronal activity. PMID- 10798395 TI - The guanine nucleotide exchange factor trio mediates axonal development in the Drosophila embryo. AB - Recent analysis of Rho subfamily GTPases in Drosophila revealed roles for Rac and Cdc42 during axonogenesis. Here, we describe the identification and characterization of the Drosophila counterpart of Trio, a guanine nucleotide exchange factor (GEF) that associates with the receptor phosphatase LAR and regulates GTPase activation in vertebrate cells. Mutants deficient in trio activity display defects in both central and peripheral axon pathways reminiscent of phenotypes observed in embryos deficient in small GTPase function. Double mutant analysis shows that trio interacts with Rac in a dose-sensitive manner but not with Rho. Moreover, reduction of trio activity potentiates the phenotype of mutations in the LAR homolog Dlar, suggesting that these proteins collaborate in orchestrating the cytoskeletal events that underlie normal axonogenesis. PMID- 10798397 TI - The Drosophila trio plays an essential role in patterning of axons by regulating their directional extension. AB - We identified the Drosophila trio gene, which encodes a Dbl family protein carrying two Dbl homology (DH) domains, each of which potentially activates Rho family GTPases. Trio was distributed along axons in the central nervous system (CNS) of embryos and was strongly expressed in subsets of brain regions, including the mushroom body (MB). Loss-of-function trio mutations resulted in the misdirection or stall of axons in embryos and also caused malformation of the MB. The MB phenotypes were attributed to alteration in the intrinsic nature of neurites, as revealed by clonal analyses. Thus, Trio is essential in order for neurites to faithfully extend on the correct pathways. In addition, the localization of Trio in the adult brain suggests its postdevelopmental role in neurite terminals. PMID- 10798396 TI - Dosage-sensitive, reciprocal genetic interactions between the Abl tyrosine kinase and the putative GEF trio reveal trio's role in axon pathfinding. AB - The Abelson tyrosine kinase (Abl) is integrated into signal transduction networks regulating axon outgrowth. We have identified the Drosophila trio gene through a mutation that exacerbates the Abl mutant phenotype. Drosophila Trio is an ortholog of mammalian Trio, a protein that contains multiple spectrin-like repeats and two Dbl homology (DH) domains that affect actin cytoskeletal dynamics via the small GTPases Rho and Rac. Phenotypic analysis demonstrates that trio and Abl cooperate in regulating axon outgrowth in the embryonic central nervous system (CNS). Dosage-sensitive interactions between trio and Abl, failed axon connections (fax), and enabled (ena) indicate that Trio is integrated into common signaling networks with these gene products. These observations suggest a mechanism by which Abl-mediated signaling networks influence the actin cytoskeleton in neuronal growth cones. PMID- 10798398 TI - Neuropeptide FF and FMRFamide potentiate acid-evoked currents from sensory neurons and proton-gated DEG/ENaC channels. AB - Acidosis is associated with inflammation and ischemia and activates cation channels in sensory neurons. Inflammation also induces expression of FMRFamidelike neuropeptides, which modulate pain. We found that neuropeptide FF (Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe amide) and FMRFamide (Phe-Met-Arg-Phe amide) generated no current on their own but potentiated H+-gated currents from cultured sensory neurons and heterologously expressed ASIC and DRASIC channels. The neuropeptides slowed inactivation and induced sustained currents during acidification. The effects were specific; different channels showed distinct responses to the various peptides. These results suggest that acid-sensing ion channels may integrate multiple extracellular signals to modify sensory perception. PMID- 10798399 TI - Structure of the Homer EVH1 domain-peptide complex reveals a new twist in polyproline recognition. AB - Homer EVH1 (Ena/VASP Homology 1) domains interact with proline-rich motifs in the cytoplasmic regions of group 1 metabotropic glutamate receptors (mGluRs), inositol-1,4,5-trisphosphate receptors (IP3Rs), and Shank proteins. We have determined the crystal structure of the Homer EVH1 domain complexed with a peptide from mGluR (TPPSPF). In contrast to other EVH1 domains, the bound mGluR ligand assumes an unusual conformation in which the side chains of the Ser-Pro tandem are oriented away from the Homer surface, and the Phe forms a unique contact. This unusual binding mode rationalizes conserved features of both Homer and Homer ligands that are not shared by other EVH1 domains. Site-directed mutagenesis confirms the importance of specific Homer residues for ligand binding. These results establish a molecular basis for understanding the biological properties of Homer-ligand complexes. PMID- 10798400 TI - Molecular basis for K(ATP) assembly: transmembrane interactions mediate association of a K+ channel with an ABC transporter. AB - K(ATP) channels are large heteromultimeric complexes containing four subunits from the inwardly rectifying K+ channel family (Kir6.2) and four regulatory sulphonylurea receptor subunits from the ATP-binding cassette (ABC) transporter family (SUR1 and SUR2A/B). The molecular basis for interactions between these two unrelated protein families is poorly understood. Using novel trafficking-based interaction assays, coimmunoprecipitation, and current measurements, we show that the first transmembrane segment (M1) and the N terminus of Kir6.2 are involved in K(ATP) assembly and gating. Additionally, the transmembrane domains, but not the nucleotide-binding domains, of SUR1 are required for interaction with Kir6.2. The identification of specific transmembrane interactions involved in K(ATP) assembly may provide a clue as to how ABC proteins that transport hydrophobic substrates evolved to regulate other membrane proteins. PMID- 10798401 TI - Constitutive activity of the light-sensitive channels TRP and TRPL in the Drosophila diacylglycerol kinase mutant, rdgA. AB - Mutations in the Drosophila retinal degeneration A (rdgA) gene, which encodes diacylglycerol kinase (DGK), result in early onset retinal degeneration and blindness. Whole-cell recordings revealed that light-sensitive Ca2+ channels encoded by the trp gene were constitutively active in rdgA photoreceptors. Early degeneration was rescued in rdgA;trp double mutants, lacking TRP channels; however, the less Ca2+-permeable light-sensitive channels (TRPL) were constitutively active instead. No constitutive activity was seen in rdgA;trpI;trp mutants lacking both classes of channel, although, like rdgA;trp, these still showed a residual slow degeneration. Responses to light were restored in rdgA;trp but deactivated abnormally slowly, indicating that DGK is required for response termination. The findings suggest that early degeneration in rdgA is caused by uncontrolled Ca2+ influx and support the proposal that diacylglycerol or its metabolites are messengers of excitation in Drosophila photoreceptors. PMID- 10798402 TI - Origin and functional impact of dark noise in retinal cones. AB - Spontaneous fluctuations in the electrical signals of the retina's photoreceptors impose a fundamental limit on visual sensitivity. While noise in the rods has been studied extensively, relatively little is known about the noise of cones. We show that the origin of the dark noise in salamander cones varies with cone type. Most of the noise in long wavelength-sensitive (L) cones arose from spontaneous activation of the photopigment, which is a million-fold less stable than the rod photopigment rhodopsin. Most of the noise in short wavelength-sensitive (S) cones arose in a later stage of the transduction cascade, as the photopigment was relatively stable. Spontaneous pigment activation effectively light adapted L cones in darkness, causing them to have a smaller and briefer dim flash response than S cones. PMID- 10798403 TI - The actions of synaptically released zinc at hippocampal mossy fiber synapses. AB - Zn2+ is present at high concentrations in the synaptic vesicles of hippocampal mossy fibers. We have used Zn2+ chelators and the mocha mutant mouse to address the physiological role of Zn2+ in this pathway. Zn2+ is not involved in the unique presynaptic plasticities observed at mossy fiber synapses but is coreleased with glutamate from these synapses, both spontaneously and with electrical stimulation, where it exerts a strong modulatory effect on the NMDA receptors. Zn2+ tonically occupies the high-affinity binding site of NMDA receptors at mossy fiber synapses, whereas the lower affinity voltage-dependent Zn2+ binding site is occupied during action potential driven-release. We conclude that Zn2+ is a modulatory neurotransmitter released from mossy fiber synapses and plays an important role in shaping the NMDA receptor response at these synapses. PMID- 10798404 TI - Calcineurin-mediated LTD of GABAergic inhibition underlies the increased excitability of CA1 neurons associated with LTP. AB - Coincident pre- and postsynaptic activity generates long-term potentiation (LTP), a possible cellular model of learning and memory. LTP has two components: (1) an increase in the excitatory postsynaptic potential (EPSP), and (2) an increase in the ability of the EPSP to generate a spike (E-S coupling of LTP). We have used pharmacological and genetic approaches to address the molecular nature of E-S coupling in CA1 pyramidal neurons. Blockade of the Ca2+-sensitive phosphatase, calcineurin, prevents induction of E-S coupling without interfering with LTP of the EPSP. Calcineurin produces its effect on E-S coupling by inducing a long lasting depression (LTD) of the GABA(A)-mediated inhibitory postsynaptic potentials (IPSPs). This LTD of the IPSP was prevented by blockade of NMDA receptors. Thus, the tetanus that elicits NMDA-dependent LTP mediates a coordinately regulated double function. It produces LTP of the EPSP and, concomitantly, LTD of the IPSP that leads to enhancement of E-S coupling. PMID- 10798405 TI - Synaptic strength as a function of post- versus presynaptic expression of the neural cell adhesion molecule NCAM. AB - To evaluate the contributions of the pre- versus postsynaptic expression of NCAM in regulation of synaptic efficacy, we cultured dissociated hippocampal cells from NCAM-deficient and wild-type mice in homo- and heterogenotypic combinations. Double recordings from synaptically coupled neurons maintained in heterogenotypic cocultures showed that synaptic strength of excitatory but not inhibitory synapses depended on expression of NCAM post- but not presynaptically. This correlated with higher levels of potentiation and synaptic coverage of NCAM expressing neurons compared to NCAM-deficient neurons in heterogenotypic cocultures. Synaptic density was the same in homogenotypic cultures of NCAM deficient and wild-type neurons as well as in heterogenotypic cocultures in which glutamate receptors were blocked. These observations indicate that the relative levels of postsynaptic NCAM expression control synaptic strength in an activity dependent manner by regulating the number of synapses. PMID- 10798406 TI - Parallel molecular pathways mediate expression of distinct forms of intermediate term facilitation at tail sensory-motor synapses in Aplysia. AB - Three distinct temporal phases of synaptic facilitation (short-, intermediate-, and long-term) are induced by serotonin (5-HT) at sensory (SN) to motor (MN) synapses in Aplysia. Here, we characterize two mechanistically distinct forms of intermediate-term facilitation (ITF) at tail SN-MN synapses. One form, activity independent ITF, is produced by five spaced pulses of 5-HT in the absence of SN activity. Its induction requires protein synthesis, and its expression requires persistent activation of PKA but not PKC. The other form, activity-dependent ITF, is produced by a single pulse of 5-HT coincident with SN activation. Its induction does not require protein synthesis, and its expression requires persistent activation of PKC but not PKA. These results demonstrate that SN-MN synapses can exhibit two distinct forms of ITF that are mediated by parallel molecular pathways. PMID- 10798407 TI - Cortical degeneration in the absence of neurotrophin signaling: dendritic retraction and neuronal loss after removal of the receptor TrkB. AB - To examine functions of TrkB in the adult CNS, TrkB has been removed from neurons expressing CaMKII, primarily pyramidal neurons, using Cre-mediated recombination. A floxed trkB allele was designed so that neurons lacking TrkB express tau-beta galactosidase. Following trkB deletion in pyramidal cells, their dendritic arbors are altered, and cortical layers II/III and V are compressed, after which there is an apparent loss of mutant neurons expressing the transcription factor SCIP but not of those expressing Otx-1. Loss of neurons expressing SCIP requires deletion of trkB within affected neurons; reduction of neuronal ER81 expression does not, suggesting both direct and indirect effects of TrkB loss. Thus, TrkB is required for the maintenance of specific populations of cells in the adult neocortex. PMID- 10798409 TI - Stability of the memory of eye position in a recurrent network of conductance based model neurons. AB - Studies of the neural correlates of short-term memory in a wide variety of brain areas have found that transient inputs can cause persistent changes in rates of action potential firing, through a mechanism that remains unknown. In a premotor area that is responsible for holding the eyes still during fixation, persistent neural firing encodes the angular position of the eyes in a characteristic manner: below a threshold position the neuron is silent, and above it the firing rate is linearly related to position. Both the threshold and linear slope vary from neuron to neuron. We have reproduced this behavior in a biophysically plausible network model. Persistence depends on precise tuning of the strength of synaptic feedback, and a relatively long synaptic time constant improves the robustness to mistuning. PMID- 10798410 TI - Sensing without touching: psychophysical performance based on cortical microstimulation. AB - Unequivocal proof that the activity of a localized cortical neuronal population provides sufficient basis for a specific cognitive function has rarely been obtained. We looked for such proof in monkeys trained to discriminate between two mechanical flutter stimuli applied sequentially to the fingertips. Microelectrodes were inserted into clusters of quickly adapting (QA) neurons of the primary somatosensory cortex (S1), and the first or both stimuli were then substituted with trains of current pulses during the discrimination task. Psychophysical performance with artificial stimulus frequencies was almost identical to that measured with the natural stimulus frequencies. Our results indicate that microstimulation can be used to elicit a memorizable and discriminable analog range of percepts, and shows that activation of the QA circuit of S1 is sufficient to initiate all subsequent neural processes associated with flutter discrimination. PMID- 10798408 TI - Role for GDNF in biochemical and behavioral adaptations to drugs of abuse. AB - The present study examined a role for GDNF in adaptations to drugs of abuse. Infusion of GDNF into the ventral tegmental area (VTA), a dopaminergic brain region important for addiction, blocks certain biochemical adaptations to chronic cocaine or morphine as well as the rewarding effects of cocaine. Conversely, responses to cocaine are enhanced in rats by intra-VTA infusion of an anti-GDNF antibody and in mice heterozygous for a null mutation in the GDNF gene. Chronic morphine or cocaine exposure decreases levels of phosphoRet, the protein kinase that mediates GDNF signaling, in the VTA. Together, these results suggest a feedback loop, whereby drugs of abuse decrease signaling through endogenous GDNF pathways in the VTA, which then increases the behavioral sensitivity to subsequent drug exposure. PMID- 10798411 TI - Congenital Heart Surgery Nomenclature and Database Project: overview and minimum dataset. AB - The International Congenital Heart Surgery Nomenclature and Database Project was organized for the purpose of standardizing nomenclature and reporting strategies that would establish the foundations for an international database. Worldwide representatives met for a series of conferences, at which time, issues of nomenclature were discussed and debated. Authors were chosen to review the various congenital heart diagnoses and reflect the mediated debate that followed. Manuscripts were prepared that reviewed the appropriate extant nomenclature, made recommendations for an inclusive rather than an exclusive method of reporting, and determined a hierarchical database scheme that would allow several levels of reporting based on the data input. This manuscript outlines two datasets for an international congenital heart surgery database, a minimum dataset and a comprehensive dataset. The comprehensive dataset includes all the imagined variables, in a hierarchical scheme, which are detailed enough to generate risk stratification analyses. The minimum dataset will include data points that would create an essential dataset, which would be mandatory for data sharing and would lend itself to basic interpretation of trends. The minimum dataset has four drop down menus for short lists of: (1) noncardiac abnormalities/general preoperative risk factors, (2) diagnoses, (3) procedures, and (4) complications, from which clinicians can choose for entry into the minimum dataset. There was universal agreement for these datasets and short lists by the assembled members of the Society of Thoracic Surgeons-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. The datasets and short lists were also unanimously approved by the Congenital Heart Surgery Committee of The European Association for Cardiothoracic Surgery and adopted by the European Congenital Heart Surgeons Foundation. PMID- 10798412 TI - Congenital Heart Surgery Nomenclature and Database Project: atrial septal defect. AB - The extant nomenclature for atrial septal defect (ASD) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798413 TI - Congenital Heart Surgery Nomenclature and Database Project: ventricular septal defect. AB - The extant nomenclature for ventricular septal defect (VSD) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Four basic VSD types are described: Subarterial, Perimembranous, Inlet, and Muscular. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analysis. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798414 TI - Congenital Heart Surgery Nomenclature and Database Project: atrioventricular canal defect. AB - The extant nomenclature for atrioventricular (AV) canal/atrioventricular septal defect is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. The three general categories are: partial AV canal (ostium primum defect), transitional (intermediate) AV canal, and complete AV canal. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798415 TI - Congenital Heart Surgery Nomenclature and Database Project: aortopulmonary window. AB - The extant nomenclature for aortopulmonary window (AP window) and pulmonary artery origin from ascending aorta (hemitruncus) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798416 TI - Congenital Heart Surgery Nomenclature and Database Project: truncus arteriosus. AB - The extant nomenclature for truncus arteriosus (TA) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A modified Van Praagh (VP) classification is proposed involving three main categories of TA: TA with confluent or near confluent pulmonary arteries (large aorta type, VP A1, A2), TA with absence of one pulmonary artery (VP A3), and TA with interrupted aortic arch or coarctation (large pulmonary artery type, VP A4). A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798417 TI - Congenital Heart Surgery Nomenclature and Database Project: pulmonary venous anomalies. AB - The extant nomenclature for pulmonary venous anomalies is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. The basis for classification are the prenatal errors of embryologic development. The major categories include: partially anomalous pulmonary venous connection, totally anomalous pulmonary venous connection, atresia of the common pulmonary vein, cor triatriatum, and stenosis or abnormal number of pulmonary veins. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Potential diagnostic-related risk factors are presented. PMID- 10798418 TI - Congenital Heart Surgery Nomenclature and Database Project: systemic venous anomalies. AB - The extant nomenclature for systemic venous anomalies is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. This nomenclature system classifies systemic venous anomalies into two primary groups by venous segment: (1) systemic venous anomalies, superior vena cava; and (2) systemic venous anomalies, inferior vena cava. Subsets are clearly defined and categorized. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798419 TI - Congenital Heart Surgery Nomenclature and Database Project: tetralogy of Fallot. AB - The extant nomenclature for tetralogy of Fallot (TOF) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. The general categories of TOF are: classic TOF with varying degrees of pulmonary stenosis, TOF with common atrioventricular canal defect, and TOF with absent pulmonary valve. Although centers may choose to code a fourth general category, TOF with pulmonary atresia, this lesion will be grouped with pulmonary atresia-ventricular septal defect for multi-institutional analysis. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798420 TI - Congenital Heart Surgery Nomenclature and Database Project: right ventricular outflow tract obstruction-intact ventricular septum. AB - The extant nomenclature for pulmonary atresia/stenosis with intact ventricular septum, pulmonary artery and/or pulmonary branch stenosis, double chambered right ventricle, absent pulmonary valve with intact ventricular septum, and ventricular to pulmonary artery conduit failure is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Multiple risk factors were considered such as coronary artery fistulas/sinusoids, tricuspid annular size and location of peripheral pulmonary artery stenoses. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Potential diagnostic related risk factors for each lesion are presented. PMID- 10798421 TI - Congenital Heart Surgery Nomenclature and Database Project: pulmonary atresia- ventricular septal defect. AB - Pulmonary atresia (PA) and ventricular septal defect (VSD) is a complex and extremely heterogeneous cardiopulmonary malformation that has not been accurately defined, as evidenced by the synonymous use of the term with tetralogy of Fallot with PA. The anatomy and morphology of the pulmonary circulation to a large extent determines the surgical approach and overall outcome, with the intracardiac anatomy playing a secondary role. Based on the characterization of the pulmonary circulation a new classification of PA-VSD is proposed. In type A, there are only native pulmonary arteries (NPA). In type B, pulmonary blood flow is provided by both NPA and by major aortopulmonary collateral arteries [MAPCA(s)]. In type C, there are only MAPCA(s) and no NPA. This new classification is proposed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798422 TI - Congenital Heart Surgery Nomenclature and Database Project: Ebstein's anomaly and tricuspid valve disease. AB - Ebstein's anomaly is a rare congenital heart defect that is characterized by a spectrum of anatomical abnormalities of the tricuspid valve that also involve the right atrium and right ventricle. The extant nomenclature for Ebstein's anomaly and our approach to the description of the severity of Ebstein's anomaly are reviewed with the objective of establishing a unified reporting system. Although there are common features in Ebstein's anomaly, there is a wide spectrum of pathology with an infinite variety of combinations of severity of the involved structures. An effort was made to develop a classification system that would take into consideration the anatomic abnormalities that help direct the surgical management, particularly in regard to tricuspid valve repair or valve replacement. Isolated congenital tricuspid stenosis and regurgitation are also rare, and a simple classification system is presented. Acquired causes of tricuspid regurgitation and stenosis are more common and are included in the classification system. A comprehensive database set for these malformations is presented so that a comprehensive risk stratification analysis can be performed. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and risk factors are presented. PMID- 10798423 TI - Congenital Heart Surgery Nomenclature and Database Project: aortic valve disease. AB - The extant nomenclature for aortic valve disease is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Aortic valve disease has been subdivided into stenotic and regurgitant lesions. Stenotic lesions have been characterized by anatomic location: supravalvar, valvar, and subvalvar. Regurgitant lesions have been characterized as either congenital or acquired. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798424 TI - Congenital Heart Surgery Nomenclature and Database Project: mitral valve disease. AB - The extant nomenclature for mitral valve disease is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Mitral valve disease has been subdivided into stenotic and regurgitant lesions. Lesions have been characterized further by etiology and by anatomic location: supravalvar, valvar, and subvalvar. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798425 TI - Congenital Heart Surgery Nomenclature and Database Project: aortic aneurysm, sinus of Valsalva aneurysm, and aortic dissection. AB - The extant nomenclature for aortic aneurysms, sinus of valsalva aneurysms, and aortic dissections is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Classification was based on morphology, histology, anatomic location, etiology, and acuity. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing that would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798426 TI - Congenital Heart Surgery Nomenclature and Database Project: aortico-left ventricular tunnel. AB - The extant nomenclature for aortico-left ventricular tunnel is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. Efforts were made to include all relevant nomenclature categories using synonyms where appropriate. The Hovaguimian classification appears most useful to surgeons: type 1, a simple tunnel with a slit-like opening at the aortic end, no aortic valve distortion; type II, a large extracardiac aortic wall aneurysm of the tunnel with an oval opening at the aortic end, with or without ventricular distortion; type III, intracardiac aneurysm of the septal portion of the tunnel, with or without right ventricular outflow tract obstruction; and type IV, a combination of type II and III. A comprehensive database set is presented, which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798428 TI - Congenital Heart Surgery Nomenclature and Database Project: pediatric cardiomyopathies and end-stage congenital heart disease. AB - The extant nomenclature for cardiomyopathy is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A functional classification based on pathophysiology is proposed. Cardiomyopathy is subdivided into: dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, right ventricular cardiomyopathy, and end-stage congenital heart disease. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798427 TI - Congenital Heart Surgery Nomenclature and Database Project: hypoplastic left heart syndrome. AB - Hypoplastic left heart syndrome (HLHS) encompasses a spectrum of structural cardiac malformations that are characterized by severe underdevelopment of the structures in the left heart-aorta complex, including the left ventricular cavity and mass. The severe end of the spectrum consists of aortic atresia and mitral atresia with a nonexistent left ventricle, whereas at the mild end patients have aortic valve and mitral valve hypoplasia without intrinsic valve stenosis, and milder degrees of left ventricular hypoplasia, recently described as hypoplastic left heart complex (HLHC). Although the overwhelming majority of the patients can only have a univentricular repair, a small minority of patients with HLHS, particularly those that are described as having HLHC, may be candidates for biventricular repair. In this paper, the extant nomenclature for HLHS is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Nomenclature and Database Committee and representatives from the European Association for Cardiothoracic Surgery. Efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A comprehensive database set is presented, which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing, and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798429 TI - Congenital Heart Surgery Nomenclature and Database Project: diseases of the pericardium. AB - The extant nomenclature for pericardial disease is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories, using synonyms where appropriate. No classification system has been reported. The groups were based on disease processes, and include: effusive pericarditis (pericardial effusion), constrictive pericarditis, cardiac tamponade, postoperative pericardial effusion, postoperative cardiac tamponade, postpericardiotomy syndrome, congenital defect, neoplastic process, benign mass, pericardial cyst, pneumopericardium, and chylopericardium. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. PMID- 10798430 TI - Congenital Heart Surgery Nomenclature and Database Project: single ventricle. AB - The extant nomenclature for single ventricle (SV) hearts is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. Efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Although many issues regarding single ventricle or univentricular hearts remain unresolved among anatomists and pathologists, a classification is proposed that is relevant to surgical therapy. A comprehensive database set is presented, which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum data set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798431 TI - Congenital Heart Surgery Nomenclature and Database Project: transposition of the great arteries. AB - The extant nomenclature for transposition of the great arteries (TGA) is reviewed for the purposes of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include relevant nomenclature categories including synonyms where appropriate. The general categories of TGA are: TGA with intact ventricular septum, TGA with ventricular septal defect (VSD) and TGA with VSD and left ventricular outflow tract obstruction (LVOTO). A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. A detailed hierarchical system is described herein for classification of the coronary artery anatomy associated with TGA. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. PMID- 10798432 TI - Congenital Heart Surgery Nomenclature and Database Project: corrected (discordant) transposition of the great arteries (and related malformations). AB - The extant nomenclature for congenitally corrected transposition of the great arteries is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Congenitally corrected transposition of the great arteries is a complex cardiac lesion that is associated with ventricular septal defect, left ventricular outflow tract obstruction, and tricuspid valve abnormalities. Nomenclature for this lesion has been variable and confusing. Herein we attempt to clarify the terminology, with an emphasis on identification of synonymous and nonsynonymous appellations. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798433 TI - Congenital Heart Surgery Nomenclature and Database Project: double outlet right ventricle. AB - Double outlet right ventricle (DORV) is a type of ventriculoarterial connection in which both great vessels arise entirely or predominantly from the right ventricle. Although the presence of aortic-mitral discontinuity and bilateral coni are important descriptors, they should not serve as absolute prerequisites for the diagnosis of DORV. The morphology of DORV is encompassed by a careful description of the ventricular septal defect (VSD) with its relationship to the semilunar valves, the great artery relationships to each other, the coronary artery anatomy, the presence or absence of pulmonary outflow tract obstruction (POTO) and aortic outflow tract obstruction (AOTO), the tricuspid-pulmonary annular distance, and the presence or absence of associated cardiac lesions. The preferred surgical treatment involves the connection of the left ventricle to the systemic circulation by an intraventricular tunnel repair connecting the VSD to the systemic semilunar valve. This ideal surgical therapy is not always possible due to the presence of confounding anatomical barriers. A multitude of alternative surgical procedures has been devised to accommodate these more complex situations. A framework for the development of the DORV module for a pediatric cardiac surgical database is proposed. PMID- 10798434 TI - Congenital Heart Surgery Nomenclature and Database Project: double outlet left ventricle. AB - Double outlet left ventricle (DOLV) is a type of ventriculoarterial connection in which both great arteries arise entirely or predominantly from the left ventricle. Although it was initially believed that bilateral absence of conus is a prerequisite for such diagnosis, all possible conal configurations have been described in this malformation. The morphology of DOLV is encompassed by a careful description of the ventricular septal defect (VSD) with its relationship to the semilunar valves, the presence or absence of pulmonary outflow tract obstruction (POTO) and aortic outflow tract obstruction (AOTO), and the presence or absence of associated cardiac lesions. The preferred surgical treatment involves the connection of the right ventricle to the pulmonary circulation by an intraventricular tunnel repair connecting the VSD to the pulmonary semilunar valve. This ideal surgical therapy is not always possible, because of the presence of confounding anatomical barriers. Several alternative surgical procedures have been devised to accommodate these more complex situations. A framework for the development of the DOLV module of a pediatric cardiac surgical database is proposed. PMID- 10798435 TI - Congenital Heart Surgery Nomenclature and Database Project: anomalies of the coronary arteries. AB - The extant nomenclature for coronary artery anomalies is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. The seven major categories of coronary artery anomalies are: anomalous pulmonary artery origins of the coronaries, anomalous aortic origins of the coronaries, congenital atresia of the left main coronary artery, coronary artery fistulas, coronary artery bridging, coronary aneurysms, and coronary stenosis. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented, which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798436 TI - Congenital Heart Surgery Nomenclature and Database Project: patent ductus arteriosus, coarctation of the aorta, interrupted aortic arch. AB - The extant nomenclature for patent ductus arteriosus (PDA), coarctation of the aorta (CoAo), and interrupted aortic arch (IAA) is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. PDA is subclassified by origin, insertion, and patient weight. CoAo is subclassified into isolated CoAo, CoAo with ventricular septal defect, and CoAo with complex intracardiac anomalies. IAA is subclassified into anatomic types A, B, and C based on the location of the interruption. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798437 TI - Congenital Heart Surgery Nomenclature and Database Project: vascular rings, tracheal stenosis, pectus excavatum. AB - The extant nomenclature for vascular rings, tracheal stenosis, and pectus deformities is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Vascular rings are subclassified as double aortic arch, right arch/left ligamentum, pulmonary artery sling, and innominate compression. Tracheal stenosis is subclassified as congenital complete tracheal rings (localized or long-segment) or acquired postintubation types. Pectus deformities are subclassified as pectus excavatum and carinatum (mild, moderate, severe). A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798438 TI - Congenital Heart Surgery Nomenclature and Database Project: arrhythmias. AB - This discussion of arrhythmia terminology attempts to classify rhythm disorders for which surgical therapy may be necessary. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery, for the purpose of establishing a unified reporting system. Efforts were made to include all relevant nomenclature categories, using synonyms where appropriate. Extant surgical ablative procedures, detailed methods of pacemaker insertion, and AICD technology are discussed. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented, which will allow for data sharing and will lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798439 TI - Congenital Heart Surgery Nomenclature and Database Project: therapeutic cardiac catheter interventions. AB - The extant nomenclature for therapeutic cardiac catheter interventions is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798440 TI - Congenital Heart Surgery Nomenclature and Database Project: end-stage lung disease. AB - The extant nomenclature for end-stage lung disease is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories, using synonyms where appropriate. Indications for lung transplantation are coded under a broad category called pulmonary failure. The proposed hierarchical scheme also allows classification of complications of lung transplantation under a category called status post lung transplant. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail, which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented, which will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented. PMID- 10798441 TI - Congenital Heart Surgery Nomenclature and Database Project: cardiac tumors. AB - The extant nomenclature for cardiac tumors is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Cardiac tumors are characterized as primary, tumors which arise directly from tissues of the heart, and secondary, tumors which arise from tissues distant from the heart, with subsequent spread to the otherwise normal tissues of the heart. Tumor types are noted in the hierarchical scheme. A comprehensive database set is presented which is based on a hierarchical scheme. Data are entered at various levels of complexity and detail which can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented which will allow for data sharing and would lend itself to basic interpretation of trends. PMID- 10798442 TI - Congenital Heart Surgery Nomenclature and Database Project: palliation, correction, or repair? AB - BACKGROUND: Inconsistent use of the terms palliation, repair, and correction, fosters unclear thinking about the goals of cardiac operation. Actual usage of these terms has not been previously documented. METHODS: Every article, for two years, pertaining to congenital heart disease in The Annals of Thoracic Surgery and The Journal of Thoracic and Cardiovascular Surgery was systematically inspected for terms of classification. RESULTS: In the 301 articles reviewed, repair and correction were used virtually interchangeably. Palliation was generally used distinctively, but all 3 descriptors were used for 3 operations. The etymology of each descriptive term suggests that they have distinct traditional definitions and connotations. Repair suggests returning to normal that which was once normal, while correction carries no implication of prior normalcy; thus, correction is the more etymologically correct term for congenital heart operations. In current literature, palliation is used improperly for a few operations to denote lack of anatomic correction or lack of permanence of correction. CONCLUSIONS: Because proper usage reflects both etymology and actual usage, we suggest that repair and correction may be used interchangeably. Palliation, however, should describe only operations that are not intended to provide normal cardiac physiology. PMID- 10798443 TI - Recycling and resensitization of delta opioid receptors. AB - Exposure to opioids results in the activation of opioid receptors; this is followed by receptor endocytosis. Previously, we showed that delta opioid receptors undergo rapid agonist-mediated internalization and that mutations in the C-tail result in a substantial loss of agonist-mediated internalization. In this study, we investigated the fate of receptors following rapid internalization. We found that the majority of the wild type receptors recycled back to the surface after acute agonist treatment. The kinetics of internalization and recycling of the receptor were virtually identical to the kinetics of internalization and recycling of the radiolabeled agonist. In contrast, the kinetics of internalization and recycling of a C-tail mutant receptor were substantially altered, suggesting an involvement of the C-tail in the recycling process. It is possible that in addition to agonist-mediated internalization, opioid receptors undergo constitutive, agonist-independent internalization. We directly examined this possibility using an antibody prebinding assay. The wild type delta opioid receptors exhibited agonist independent internalization via the clathrin-coated pit pathway. We also examined the role of receptor internalization and recycling in the modulation of its function by quantitating the level of opioid-stimulated phosphorylation of MAP kinase (MAPK) under conditions of receptor internalization and recycling. We found that agonist treatment caused a rapid increase in the level of phosphorylated MAPK that was rapidly desensitized. The removal of the agonist, which results in receptor recycling, led to the resensitization of the receptor, as evidenced by the agonist's ability to reinduce MAPK phosphorylation. Mutant receptors that underwent rapid recycling exhibited enhanced resensitization, suggesting a role for receptor recycling in the resensitization process. Taken together, these results indicate that agonist-mediated internalization and recycling modulate opioid receptor function and that the receptor C-tail plays an important role in both processes. PMID- 10798445 TI - Characterization of a subcloned fragment (pBA0.6) of pCMM86 located on 17q21 and its potential use in generating an individual-specific DNA profile. AB - Sequence analysis was carried out of a human clone pBA0.6 generated after exonuclease III/S1 nuclease digestion and subcloning of pCMM86 (GDB: 168382, D17S74), which was not available in the database. It revealed the presence of a reiterating core motif of 24mer GTGGGTGTGTTGGAGGGGGTGAGG, present 23 times, which was GC-rich and minisatellitic in nature. Genomic blots of HaeIII-digested human DNA, when hybridized with pBA0.6, generated a ladder of bands between 29.0 kb and 2.1 kb. Hybridization analyses of 88 unrelated individuals belonging to four regions of India using this probe revealed polymorphic bands which were individual specific. The probability of identity ranged from 5.07x10(-14) in Punjabis to 2.64x10(-16) in Bengalis and was found to be 3.06x10(-16) in UPites, whereas in the case of South Indians, it was 3.9x10(-15). Three sets of isomorphic bands at 29.0 kb, 2.4 kb, and 2.1 kb were common between the individuals of all the regions and served as internal markers. The 29.0-kb band was observed to be Homo sapiens specific. Construction of dendrograms based on the UPGMA method with Jaccard's coefficient values suggested less genetic similarity/high genetic diversity in all the population groups, indicating that the samples taken were random. Maximum likelihood estimates through the bootstrap sampling method showed that Punjabis, Bengalis, and UPites formed one cluster, whereas South Indians formed a separate cluster, altogether thus showing the proximity of these three population groups compared with that from South India. A preliminary study by Northern hybridization with pBA0.6 resulted in two transcripts of 0.63 kb and 0.29 kb. This finding was corroborated with RT-PCR results where 2 amplicons, matching the expected size of two open reading frames within the minisatellite sequence, were obtained. The role of the two transcripts from the minisatellite sequence is not clear as yet, and it is probable that these messages may not get translated because of the absence of a eukaryotic Kozak sequence around the initiator methionine in the pBA0.6 sequence. PMID- 10798444 TI - Gene structure and chromosomal assignment of mouse GITR, a member of the tumor necrosis factor/nerve growth factor receptor family. AB - GITR is a type I transmembrane protein that belongs to the tumor necrosis factor/nerve growth factor receptor (TNF/NGFR) family. This receptor is preferentially expressed in activated T lymphocytes and may function as signaling molecule during T-cell development. In the present study, we examined the genomic organization of the entire mouse GITR (mGITR) gene. The gene spans a 2543-bp region and consists of five exons (with a length ranging from 88 bp to 395 bp) and four introns (67 bp to 778 bp). In agreement with GITR expression in activated T cells, consensus elements for transcription factors involved in T cell development and activation were identified in the 5' flanking region, including a consensus element for NF-kappaB. Two highly significant binding sites for MyoD and one binding site for myogenin were also found, suggesting involvement of GITR in muscle development. The mGITR gene contains 17 transcription initiation sites distributed over a 76-bp region, all used with the same frequency. We localized mGITR to the murine chromosome 4 (E region), where other 4 TNF/NGFR members localize, including m4-1BB and mOX40. These results further indicate that GITR shares several features with OX40, 4-1BB, and CD27, suggesting the existence of a new subfamily of the TNFR family, as also confirmed by the similarity of their cytoplasmic domains. PMID- 10798446 TI - Characterization of a zinc finger protein ZAN75: nuclear localization signal, transcriptional activator activity, and expression during neuronal differentiation of P19 cells. AB - The ZAN75 cDNA was first identified in NIH 3T3 cells and codes for a DNA-binding protein with two zinc finger motifs. In this study, we characterized the nuclear localization signal of ZAN75, tested if ZAN75 regulates transcription, and examined its expression during embryonic development and neuronal differentiation of P19 mouse embryonal carcinoma cells. By examining the cellular localization of deletion mutants of ZAN75 fused to green fluorescence protein, ZAN75 was revealed to have a bipartite nuclear localization signal sequence upstream of the zinc finger domains. The N-terminal region of ZAN75, when fused to the GAL4 DNA binding domain, strongly activated transcription. The expression of ZAN75 mRNA was found to be developmentally regulated, showing the highest expression in E11.5 embryos. In situ hybridization experiments using E11.5 embryos showed a high expression of the transcripts in neuronal tissues such as brain and neural tube. The expression of ZAN75 was transiently increased at both the mRNA and the protein levels when P19 cells were treated with retinoic acid to induce neuronal differentiation. Taken together, these results indicate that ZAN75 is a transcriptional activator with a bipartite nuclear localization signal and may play a role in neuronal differentiation. PMID- 10798447 TI - A triplex-mediated knot between separated polypurine-polypyrimidine tracts in circular DNA blocks transcription by Escherichia coli RNA polymerase. AB - Polypurine-polypyrimidine tracts are overrepresented in eukaryotes and many have the potential to form triplex DNA. Transmolecular triplexes form between separated but complementary polypurine-polypyrimidine tracts in duplex DNA. Transmolecular triplexes (T-loops) were studied previously using a circular plasmid containing a pair of separated polypurine-polypyrimidine tracts designed to able to form a triplex with each other. T-Loops formed when the nicked plasmid was incubated at low pH in the presence of spermine. When the pH was raised to 8, the T-loops were constrained by a hydrogen-bonded knot composed of multistranded and single-stranded regions. The present experiments used T-loops as a model system to investigate the influence of transmolecular triplex formation on transcription. T-Loops and control open circular, linear, and supercoiled plasmid forms were isolated from bands on agarose gels. Transcription assays were carried out with the isolated plasmid forms and Escherichia coli RNA polymerase holoenzyme and the core enzyme, which lacked sigma70. Transcription was significantly inhibited in T-loop forms compared with control plasmid forms. There was no evidence that the single-stranded regions of T-loops facilitated nonspecific initiation of transcription. Instead, the multistranded component of the hydrogen-bonded knot at the root of the T-loop structure inhibited transcription. PMID- 10798448 TI - Murine monoclonal antibodies biologically active against the amino region of HIV 1 gp120: isolation and characterization. AB - The human immunodeficiency virus (HIV)-1 envelope glycoprotein is synthesized as a precursor (gp160) and subsequently cleaved to generate the external gp120 and transmembrane gp41 glycoproteins. Both gp120 and gp41 have been demonstrated to mediate critical functions of HIV, including viral attachment and fusion with the cell membrane. The antigenic variability of the HIV-1 envelope glycoprotein has presented a significant problem in the design of appropriate and successful vaccines and offers one explanation for the ability of HIV to evade immune surveillance. Therefore, the development and characterization of functional antibodies against conserved regions of the envelope glycoprotein is needed. Because of this need, we generated a panel of murine monoclonal antibodies (MuMabs) against the HIV-1 envelope glycoprotein. To accomplish this, we immunized Balb/C mice with a recombinant glycoprotein 160 (gp160) that was synthesized in a baculovirus expression system. From the growth-positive hybridomas, three MuMabs were generated that demonstrated significant reactivity with recombinant gp120 but failed to show reactivity against HIV-1 gp41, as determined by enzyme-linked immunosorbent assay (ELISA). Using vaccinia constructs that synthesize variant truncated subunits of gp160, we were able to map reactivity of all three of the Mabs (ID6, AC4, and AD3) to the first 204 residues of gp120 (i.e., the N terminus of gp120) via Western blot analysis. Elucidation of the epitopes for these Mabs may have important implications for inhibition of infection by HIV-1. Our initial attempts to map these Mabs with linear epitopes have not elucidated a specific antigenic determinant; however, several physical characteristics have been determined that suggest a continuous surface epitope. Although these antibodies failed to neutralize cell-free or cell associated infection by HIV-1, they did mediate significant antibody-dependent cellular cytotoxicity (ADCC) activity, indicating potential therapeutic utility. In summary, these data suggest the identification of a potentially novel site in the first 200 aa of gp120 that mediates ADCC. PMID- 10798449 TI - Some basic issues for clinicians concerning things statistical. PMID- 10798450 TI - The 1999 IPA/Bayer Research Awards. PMID- 10798451 TI - Quality adjusted life years in older adults with depressive symptoms and chronic medical disorders. AB - We used data from a 4-year prospective study of 2,558 primary care patients age 65 and older in a large staff model health maintenance organization to examine the association of clinically significant depressive symptoms and eight other chronic medical conditions with quality adjusted life years (QALYs). We developed linear regression models to examine the association of clinically significant depressive symptoms as defined by a score of 16 or greater on the Center for Epidemiological Studies Depression Scale and eight common chronic medical disorders at baseline with QALYs over the 4-year study period. Estimates of QALYs were derived from Quality of Well-Being Scale scores at baseline, at 2-year follow-up, and at 4-year follow-up. Individuals with clinically significant depressive symptoms at baseline had significantly lower QALYs over the 4-year study period than nondepressed subjects, even after adjusting for differences in age, gender, and the eight other chronic medical conditions. In terms of the entire study population, only arthritis and heart disease were more strongly associated with QALYs than depression. PMID- 10798452 TI - Geriatric psychiatric morbidity in rural northern India: implications for the future. AB - This was an epidemiological study funded by the Indian Council of Medical Research and conducted in a representative rural geographical area in northern India. The study aimed to determine the pattern of psychiatric morbidity in those aged 60 years or above and to ascertain the causative/contributory role of biosociodemographic factors, if any. Three groups of subjects formed the sample: the geriatric psychiatrically ill and non-ill and the nongeriatric psychiatrically ill. The prevalence of psychiatric morbidity was found to be much higher in the geriatric group (43.32%) than in the nongeriatric group (4.66%). The psychiatric morbidity pattern consisted of neurotic depression, manic depressive psychosis depression, and anxiety state in descending order of frequency. Socially, economically, and educationally disadvantaged subjects were found to be more psychiatrically ill. The data indicate that by the year 2000, there will be about 28 million psychiatrically ill geriatric persons in the country requiring the attention of health policy planners. The implications of these findings in light of the available services for psychiatrically ill geriatric persons have been discussed and future directions identified. PMID- 10798453 TI - Effects of individualized versus classical "relaxation" music on the frequency of agitation in elderly persons with Alzheimer's disease and related disorders. AB - Confusion and agitation in elderly patients are crucial problems. This study tested Gerdner's mid-range theory of individualized music intervention for agitation. An experimental repeated measures pretest-posttest crossover design compared the immediate and residual effects of individualized music to classical "relaxation" music relative to baseline on the frequency of agitated behaviors in elderly persons with Alzheimer's disease and related disorders (ADRD). Thirty nine subjects were recruited from six long-term-care facilities in Iowa. The sample consisted of 30 women and 9 men (mean age 82 years) with severe cognitive impairment. Baseline data were collected for 3 weeks. Findings from the Modified Hartsock Music Preference Questionnaire guided the selection of individualized music. Group A (n = 16) received individualized music for 6 weeks followed by a 2 week "washout" period and 6 weeks of classical "relaxation" music. Group B (n = 23) received the same protocol but in reverse order. Music interventions were presented for 30 minutes, two times per week. The Modified Cohen-Mansfield Agitation Inventory measured the dependent variable. A repeated measures analysis of variance with Bonferroni post hoc test showed a significant reduction in agitation during and following individualized music compared to classical music. This study expands science by testing and supporting a theoretically based intervention for agitation in persons with ADRD. PMID- 10798454 TI - Dementia and depression in elderly medical inpatients. AB - The purpose of this study was to examine the prevalence and correlation of cognitive impairments, major depression, and depressive symptoms among elderly medical inpatients, and to compare the degree of depressive symptomatology as well as cognitive deterioration in possible vascular dementia and possible Alzheimer's disease. In a department of internal medicine, 100 (36 male, 64 female) 65-year-old or older patients were examined by a semistructured interview, and assessed by the Hachinski Ischemic Scale, the Hamilton Rating Scale for Depression (HDS), and the Modified Mini-Mental State (MMMS) Examination. In our total sample, the MMMS total score was (+/-SD) 76.0+/-15.5 and the HDS total score was (+/-SD) 12.0+/-6.1. Based on DSM-IV criteria, major depression was established in 11 patients. Deterioration of cognitive functions was seen in 66 patients; cognitive impairment was mild in 30 patients, moderate in 19, and severe in 17. Forty-six patients had mild depressive symptoms and 27had severe depressive symptoms. In summary, a high prevalence of cognitive dysfunction and depressive symptomatology was detected in our study, illustrating the importance of psychiatric care in elderly medical inpatients. PMID- 10798455 TI - Coping with depression and anxiety: preliminary results of a standardized course for elderly depressed women. AB - The experiences of 51 elderly depressed female patients with a standardized course "Coping with Depression and Anxiety" were investigated. Preliminary findings concerning the effectiveness of this course in 34 patients who completed the course will also be presented. This course consisted of psychoeducation and skills training. Depressive symptoms were scored by using the SCL-90 depression scale and the Geriatric Depression Scale. Results indicate that patients were very enthusiastic about the course and that there was no need to make major changes in its structure and content. Also, depressive symptoms were significantly reduced after completion of the course, which emphasizes the importance of this psychoeducational program in the treatment of depression in elderly female patients. PMID- 10798456 TI - Determinants and detection of low body mass index in community-dwelling adults with Alzheimer's disease. AB - OBJECTIVE: To determine the proportion of older adults with Alzheimer's disease presenting to a geriatric clinic with low body mass index (BMI), the proportion of these individuals recognized by clinicians as malnourished, and what patients' characteristics and caregivers' and clinicians' impressions are associated with low BMI. DESIGN: Cross-sectional study. SETTING: An outpatient geriatric clinic located in a university-affiliated teaching hospital. PARTICIPANTS: 340 patients with Alzheimer's disease, average age 75 years. MEASUREMENTS: Individuals with a BMI below 21 were considered at risk of malnutrition. Physical examination and medical information were obtained from patients and caregivers by clinicians using a standardized assessment protocol. Clinicians' impression regarding evidence of malnutrition was obtained. RESULTS: Forty-six patients (16%) had a BMI below 21. Clinicians reported evidence of potential malnutrition in 11 patients, 8 of whom had a BMI below 21. Using logistic regression, we found that women were five times more likely to have a BMI below 21 than men, and that individuals with low cognition were twice as likely to have a BMI below 21 than individuals with higher cognition. CONCLUSION: The proportion of patients with Alzheimer's disease with a BMI below 21 is similar to that encountered in the general population aged 65+. However, clinicians have difficulty identifying persons at risk of malnutrition according to BMI status. Women with low cognition were at increased risk of having a low BMI. Improvement in the detection of malnutrition is desirable. Further exploration of causal links between cognition and malnutrition is required. PMID- 10798457 TI - Effect of integrated family support versus day care only on behavior and mood of patients with dementia. AB - OBJECTIVE: This study tests the hypothesis that integrated family support, in which patients and caregivers are both supported by one professional staff, is more effective in influencing behavior problems and mood of the dementia patient than nonintegrated support, such as psychogeriatric day care only. DESIGN: A quasi-experimental pretest-posttest control group design with matched groups was applied. SETTING: Psychogeriatric day-care centers of four community centers and three nursing homes. PARTICIPANTS: Fifty-six dementia patients living at home and their caregivers. INTERVENTION: The patients in the experimental group (n = 33) participated together with their caregivers in an integrated family support program, whereas the patients in the control group (n = 23) received psychogeriatric day care only. MEASUREMENTS: Behavior problems and mood were observed using standardized behavior observation scales. RESULTS: After 7 months the experimental support program, compared to the regular psychogeriatric day care, showed a large positive effect on the total number of behavior problems (effect size .75), and also specifically on the degree of inactivity (effect size .66) and nonsocial behavior (effect size .61). No effect on mood was found. CONCLUSIONS: In influencing the total amount of behavior problems, as well as the degree of inactivity and nonsocial behavior, the integrated family support program proved to be more effective than psychogeriatric day care. Because behavior problems are an important determinant for admission of persons with dementia into a nursing home, integrated family support may contribute to the delay of institutionalization. PMID- 10798458 TI - Development of delirium: a prospective cohort study in a community hospital. AB - BACKGROUND: Previous research on risk of delirium in acute hospital settings identified mainly patient variables (e.g., age) that are not amenable to intervention. The purpose of this study was to develop a model for new delirium in hospitalized older patients that included process of care and social variables. METHODS: A prospective cohort study was undertaken in a community hospital in Ontario, Canada. Research participants included 156 hospitalized patients age 65+ years and without delirium on admission who were admitted to a medical or surgical unit. The measures included daily appraisal of delirium using a standardized and validated tool, and assessment of patient, process of care, and social variables. RESULTS: Delirium developed in 28 of the 156 patients (17.9%). Older age and cognitive impairment were significant patient variables. Significant process of care variables included a high number of medications administered during hospitalization, surgery, a high number of procedures during early hospitalization (e.g., x-rays, blood tests), and intensive care treatment. CONCLUSIONS: Approximately one older patient in five developed delirium after admission to a medical or surgical unit. Risks not easily amenable to intervention included age, cognitive dysfunction, surgery, and intensive care requirements. Risk factors that are potentially modifiable included number of medications and number of procedures. Future research might focus on the efficacy of such intervention to reduce new-onset delirium in acute hospital settings. PMID- 10798459 TI - Change in disability follows inpatient treatment for late life depression. AB - OBJECTIVES: The objective of this study was to examine the relationship between functional disability and improvement in late life depression after acute inpatient treatment. DESIGN: The study was a longitudinal assessment of depression and disability. Patients were assessed during an initial inpatient hospitalization and then 3 months postdischarge. SETTING: All patients were evaluated initially after admission to one of 71 inpatient psychiatric treatment facilities. PARTICIPANTS: The study comprised of 2572 patients older than age 60 who were relatively cognitively intact and experiencing significant depressive symptoms. MEASUREMENTS: Depressive symptoms were measured using the Geriatric Depression Scale. Disability was measured using the Instrumental Activities of Daily Living Scale and the Medical Outcomes SF-36. RESULTS: Depressive symptoms improved in the majority of patients. Moreover, improvement in depressive symptomatology was significantly related to improvement in instrumental activities of daily living (IADLs) and to health-related quality of life as measured by the SF-36. This relationship was strongest among those who initially presented with some disability in IADLs. CONCLUSIONS: This work underscores further the disabling nature of depression. Moreover, findings from this study suggest that treatment focused on depression can lead to significant improvements in both depressive symptoms and functional abilities. However, the results also suggest that the relationship between depression and disability is complex and that the effect of treating depression is not the only factor in the reversal of disability. PMID- 10798460 TI - Selecting target conditions for quality of care improvement in vulnerable older adults. AB - OBJECTIVE: To identify a set of geriatric conditions as optimal targets for quality improvement to be used in a quality measurement system for vulnerable older adults. DESIGN: Discussion and two rounds of ranking of conditions by a panel of geriatric clinical experts informed by literature reviews. METHODS: A list of 78 conditions common among vulnerable older people was reduced to 35 on the basis of their (1) prevalence, (2) impact on health and quality of life, (3) effectiveness of interventions in improving mortality and quality of life, (4) disparity in the quality of care across providers and geographic areas, and (5) feasibility of obtaining the data needed to test compliance with quality indicators. A panel of 12 experts in geriatric care discussed and then ranked the 35 conditions on the basis of the same five criteria. We then selected 21 conditions, based on panelists' iterative rankings. Using available national data, we compiled information about prevalence of the selected conditions for community-dwelling older people and older nursing home residents and estimated the proportion of inpatient and outpatient care attributable to the selected conditions. RESULTS: The 21 conditions selected as targets for quality improvement among vulnerable older adults include (in rank order): pharmacologic management; depression; dementia; heart failure; stroke (and atrial fibrillation); hospitalization and surgery; falls and mobility disorders; diabetes mellitus; end-of-life care; ischemic heart disease; hypertension; pressure ulcers; osteoporosis; urinary incontinence; pain management; preventive services; hearing impairment; pneumonia and influenza; vision impairment; malnutrition; and osteoarthritis. The selected conditions had mean rank scores from 1.2 to 3.8, and those excluded from 4.6 to 6.9, on a scale from 1 (highest ranking) to 7 (lowest ranking). Prevalence of the selected conditions ranges from 10 to 50% among community-dwelling older adults and from 25 to 80% in nursing home residents for the six most common selected conditions. The 21 target conditions account for at least 43% of all acute hospital discharges and 33% of physician office visits among persons 65 years of age and older. Actual figures must be higher because several of the selected conditions (e.g., end-of-life care) are not recorded as diagnoses. CONCLUSIONS: Twenty-one conditions were selected as targets for quality improvement in vulnerable older people for use in a quality measurement system. The 21 geriatric conditions selected are highly prevalent in this group and likely account for more than half of the care provided to this group in hospital and ambulatory settings. PMID- 10798461 TI - Combined behavioral and drug therapy for urge incontinence in older women. AB - OBJECTIVE: The purpose of this study was to examine the effects of combining behavioral treatment and drug treatment for urge incontinence in community dwelling older women. DESIGN: Modified crossover design (extension of a randomized clinical trial). Eligible subjects were stratified according to type and severity of incontinence and randomized to behavioral treatment, drug treatment, or a control condition (placebo). Subjects not totally continent or not satisfied after 8 weeks of a single treatment were offered the opportunity to cross over into combined therapy. SETTING: A university-based outpatient geriatric medicine clinic. PARTICIPANTS: Subjects in the clinical trial were 197 ambulatory, nondemented, community-dwelling women (age 55 years or older) with persistent urge urinary incontinence. Thirty-five subjects participated in combined treatment. INTERVENTION: One group of subjects received four sessions (over 8 weeks) of biofeedback-assisted behavioral training followed by 8 weeks of behavioral training combined with drug therapy (oxybutynin chloride individually titrated from 2.5 mg to 15 mg daily). The second group received drug therapy first, followed by 8 weeks of drug therapy combined with behavioral training. MEASUREMENTS: Bladder diaries completed by subjects before and after each treatment phase were used to calculate change in the frequency of incontinent episodes. RESULTS: Eight subjects (12.7%) crossed from behavioral treatment alone to combined behavioral and drug therapy. Additional benefit was seen in improvement from a mean 57.5% reduction of incontinence with single therapy to a mean 88.5% reduction of incontinence with combined therapy (P = .034). Twenty seven subjects (41.5%) crossed from drug therapy alone to combined drug and behavioral treatment. They also showed additional improvement, from a mean 72.7% reduction of incontinence with single therapy to a mean 84.3% reduction of incontinence with combined therapy (P = .001). CONCLUSIONS: This study shows that combining drug and behavioral therapy in a stepped program can produce added benefit for patients with urge incontinence. PMID- 10798462 TI - Binocular vision in older people with adventitious visual impairment: sometimes one eye is better than two. AB - PURPOSE: To determine the effect of adventitious visual impairment (low vision) on monocular and binocular spatial contrast sensitivity of older people. DESIGN: A between-within repeated measures design. PARTICIPANTS: Fifty-nine older adults between the ages of 50 and 96; 49 of the observers had age-related macular degeneration (AMD). MEASURES: Visual acuity screening and spatial contrast sensitivity. RESULTS: In almost half of the population with AMD, the sensitivity to spatial information, as measured by spatial contrast sensitivity, is worse when both eyes are used than when the stimuli are viewed with only one eye. This "binocular inhibition" is not related to the contrast sensitivity of the better eye or to acuities. Furthermore, this inhibition process is reflected primarily in images with medium to low spatial frequency components (medium to large size bars). CONCLUSIONS: These results have important implications for understanding the functional impact of low vision in older people. They suggest that almost one half of older people with AMD view the world best using only one of their eyes, whereas for the other half, there is an advantage to using binocular vision for certain visual tasks. PMID- 10798463 TI - Skeletal muscle mass and muscle strength in relation to lower-extremity performance in older men and women. AB - OBJECTIVE: Low muscle strength is associated with poorer physical function, but limited empirical evidence is available to prove the relationship between muscle mass and physical function. We tested the hypothesis that persons with lower muscle mass or muscle strength have poorer lower-extremity performance (LEP). DESIGN: A cross-sectional, population-based study. PARTICIPANTS: A cohort of 449 men and women aged 65 years and older living in Amsterdam and its surroundings participating in the second examination (1995-1996) of the Longitudinal Aging Study Amsterdam. MEASUREMENTS: Leg skeletal muscle mass was measured using dual energy X-ray absorptiometry (DXA). Grip strength was used as an indicator of muscle strength. Timed functional performance tests, including walking and repeated chair stands, were used to assess LEP. RESULTS: After adjustment for body height and age, leg muscle mass was positively associated with LEP in men (regression coefficient 0.178 [95% confidence interval 0.013-0.343], P = .035). In women an inverse association was observed, which became positive after additional adjustment for body mass index (BMI) (0.202 [-0.001-0.405], P = .052). Grip strength was positively associated with LEP in men and women. After additional adjustment for behavioral, physiological, and psychological variables, the associations between leg muscle mass and LEP disappeared, whereas grip strength remained to be independently associated with LEP in men (0.079 [0.042 0.116], P = .0001), with a tendency in women (0.046 [-0.009-0.101], P = .11). Results were similar when quartiles of leg muscle mass or grip strength were used. CONCLUSIONS: These results suggest that low muscle strength, but not low muscle mass, is associated with poor physical function in older men and women. However, prospective studies are needed to investigate the association between loss of muscle mass and physical function. PMID- 10798464 TI - Atrial fibrillation and cognitive disorders in older people. AB - OBJECTIVES: To find a correlation between chronic nonrheumatic atrial fibrillation (CNRAF) and cognitive impairment in a group of older, nondemented patients. SETTING: Acute Care Unit for the Elderly, Poliambulanza Hospital, Brescia (Italy). METHODS: Two hundred fifty-five hospital in-patients older than 70 years (42 with CNRAF and 213 controls with normal sinus rhythm) were assessed by complete clinical history, physical examination, ECG, serum albumin levels, APACHE II score, mental status (Mini-Mental State Exam [MMSE] and Geriatric Depression Score [GDS]), functional status (Barthel Index and instrumental activities of daily living [IADL]), number of prescribed drugs, and comorbidity (Charlson Index). RESULTS: The group of patients with CNRAF had MMSE scores significantly lower than that of the reference group with normal sinus rhythm. Chronic nonrheumatic atrial fibrillation retained an independent relation to cognitive impairment also after adjusting for those variables associated with mental decline in univariate models (GDS, IADL, and APACHE II scores). CONCLUSIONS: The results of this study support the relationship between nonrheumatic atrial fibrillation and impaired cognitive function. Independent of etiopathogenetic mechanisms (thromboembolic or hemodynamic hypotheses), prevention of cognitive impairment in older persons should take into account the treatment of atrial fibrillation and its consequences. PMID- 10798465 TI - A cross-national survey of tube-feeding decisions in cognitively impaired older persons. AB - OBJECTIVES: Many factors affect the decision to institute long-term tube-feeding in older persons. The objectives of this cross-national survey are to examine the tube-feeding decision-making process for cognitively impaired older persons from the perspective of the substitute decision-makers (SDM) and to contrast this process in US and Canadian healthcare settings. DESIGN: Survey. SETTING: Chronic care facilities in Ottawa and nursing homes in Boston. PARTICIPANTS: Patients more than age 65 who were tube-fed for at least 2 months and who were unable to make their own healthcare decisions at the time of tube placement were identified at both sites. The SDMs of 46 patients in Ottawa and 48 patients in Boston were surveyed. MEASUREMENTS: The survey asked questions relating to the following categories: health status of the patient, advance directives, communication with the healthcare team, perceived goals of tube-feeding, decision satisfaction, and sociodemographic data. RESULTS: Tube-fed patients in Boston were more likely to have a diagnosis of dementia than those in Ottawa (60.4% vs 10.9%, P < .001) and were less likely to have had an acute neurological event (35.4% vs 71.7%, P < .001). There was a greater likelihood in Boston than in Ottawa (68.7% vs 6.5%, P < .001) for tube-feeding decisions to be made in a nursing home (vs an acute hospital). In the combined cohort, 19.1% of patients had a living will, and only 47.9% of SDMs felt confident that the patients would want to have a feeding tube. The majority of SDMs at both sites felt they understood the benefits (83.0%), but not the risks (48.9%), of tube-feeding. The most commonly perceived reasons for tube-feeding were to "prolong life" (84.0%) and to "prevent aspiration" (67.0%). Approximately half of all SDMs felt they had received adequate support from the healthcare team. A minority of SDMs (38.3%) at both sites stated that they would want a feeding tube for themselves, and only 40% of SDMs felt the feeding tube had improved the patients' quality of life. CONCLUSIONS: A greater proportion of patients have feedings tubes inserted because of a degenerative dementia in Boston compared with an acute neurological event in Ottawa. Despite the difference in diagnostic indication for tube-feeding, the substitute decision making process was seriously limited at both sites by poor implementation of the principle of substituted judgement, a need for broader advance directives, and improved transfer of knowledge between clinicians and decision-makers. PMID- 10798466 TI - A drug use evaluation of selected opioid and nonopioid analgesics in the nursing facility setting. AB - OBJECTIVES: To determine the medical conditions for which selected analgesics are most frequently prescribed in nursing facilities (NFs), describe the use of pharmacologic and nonpharmacologic pain therapies, and determine the frequency and quality of pain assessment in NF residents. DESIGN: A multicenter, 3-month retrospective drug use evaluation conducted by consultant pharmacists. SETTING: Eighty-nine NFs having no more than 25% of their patient census representing special populations (e.g., head trauma). PARTICIPANTS: A total of 2065 adult NF residents who received at least one selected analgesic. MEASUREMENTS: Primary indication for analgesics, pain type, method of pain assessment, nonpharmacologic therapies for pain, prescribed analgesics and regimens, and comorbid conditions were recorded. RESULTS: A total of 54.3% of residents had one indication for analgesic therapy, 31.0% had two indications, and 14.7% had three or more indications. Arthritis was the most prevalent indication for analgesics (41.7% of residents), followed by bone fracture (12.4%) and other musculoskeletal conditions (9.7%). More residents (76.8%) were reported to have chronic pain than acute pain (19.9%), and 3.0% had both chronic and acute pain. Pain type was unknown for 0.2% of residents. Observational pain assessments were used more frequently (for 55.9% of residents) than objective methods (16.6%), and pain was not assessed in 40.6% of residents. Most residents (69.4%) received no nonpharmacologic treatment for pain. Of the 2542 opioid and nonsteroidal anti inflammatory drug (NSAID) prescriptions, 67.6% were for opioids, 24.8% were for NSAIDs, and 7.6% were for tramadol. Propoxyphene-containing drugs were the most frequently prescribed opioid group, and propoxyphene with acetaminophen was the most frequently prescribed analgesic (35.6% of all analgesics). Most analgesics (63.2%) were prescribed on an as-needed (prn) basis. CONCLUSIONS: The findings show a lack of adequate pain assessments, little use of nonpharmacologic interventions, and inappropriate use of analgesic medication. The small percentage of residents with chronic pain assessed objectively suggests the difficulty of monitoring pain progression in NFs. The prescribing of analgesic for most residents (with propoxyphene used most often, long-acting opioids used infrequently, and frequent prn use) was inconsistent with recommended pain therapy in older people and attests to the urgent need to educate NF practitioners on the appropriate use of analgesics. PMID- 10798467 TI - Predicting adverse postoperative outcomes in patients aged 80 years or older. AB - OBJECTIVE: The identification of reversible factors that are associated with postoperative morbidity in geriatric surgical patients is critical to improving perioperative outcomes in such patients. Our study aimed to compare the relative importance of intraoperative versus preoperative factors in predicting adverse postoperative outcomes in geriatric patients. DESIGN: Retrospective cohort study of consecutive patients undergoing noncardiac surgery in 1995. SETTING: Two University of California, San Francisco, teaching hospitals--Moffitt/Long and Mount Zion medical centers. PARTICIPANTS: All men and women 80 years of age or older undergoing noncardiac surgery. MEASUREMENTS: Medical records of all patients were reviewed to measure predefined pre- and intraoperative risk factors and postoperative outcomes. Predictors of postoperative outcomes were identified by multivariate logistic regression analyses. RESULTS: Three hundred sixty-seven patients were studied. The most prevalent preoperative risk factors were a history of hypertension and coronary artery, pulmonary, and neurologic diseases. Postoperative in-hospital mortality rate was 4.6%, and 25% of patients developed adverse postoperative outcomes, of which neurological and cardiovascular complications were the leading causes of morbidity (15% and 12%, respectively). By multivariate logistic regression, a history of neurological disease (odds ratio [OR] 4.0, 95% confidence interval [CI] 2.3 - 6.9, P = .0001), congestive heart failure (OR 2.7, 95% CI 1.4 - 5.3, P = .004), and a history of arrhythmia (OR 2.3, 95% CI 1.2 - 4.3, P = .01) increased the odds of adverse postoperative events. The only intraoperative event shown to be predictive of postoperative complications was the use of vasoactive agents (OR 8.0, 95% CI 1.6 - 40.5, P = .009). CONCLUSIONS: In this group of geriatric surgical patients, the overall postoperative in-hospital mortality rate was 4.6%, and 25% of the patients developed adverse postoperative outcomes involving either the neurological, cardiovascular, or pulmonary systems. Intraoperative events appeared to be less important than preoperative comorbidities in predicting adverse postoperative outcomes. PMID- 10798468 TI - The association of depressive symptoms and urinary incontinence among older adults. AB - OBJECTIVES: To examine the relationship of urinary incontinence (UI) and depressive symptoms (DS) in older adults. DESIGN: A randomized, controlled trial to determine the effects of clinical practice guideline implementation on provider attitudes and behavior, and patients' UI, health status, quality of life, and satisfaction with care. Baseline and endpoint data were collected from patients via computer-assisted telephone interviewing. SETTING: Forty-one nonacademic primary care practices (PCP) in North Carolina. PARTICIPANTS: A total of 668 community-dwelling adults (age > 60) who had visited the one of the selected PCPs. INTERVENTION: PCPs in the intervention group were given instruction in the detection and management of UI, educational materials for providers and patients, office system supports, and academic detailing. MEASUREMENTS: The dependent measure was assessed using an eight-item screener for DS. UI (status, frequency, amount), health (physical, mental), and demographic (age, gender, marital status) and self-report information about bladder control served as predictors. RESULTS: Wilcoxon rank sum tests showed that UI status was associated with moderate to severe DS (43% vs 30%, P = .05). Multivariate analyses showed that UI status, physical and mental health, and gender were significant predictors of DS. Among UI adults (n = 230), physical and mental health, life satisfaction, and the perception that UI interfered with daily life were significant predictors of DS. CONCLUSIONS: This study provides clear evidence that UI is related to DS in older adults. PMID- 10798469 TI - Health, functional, and psychological outcomes among older persons with chronic dizziness. AB - OBJECTIVE: The recommended clinical strategy for a health condition should depend both on the known causes of and outcomes associated with the condition. The aim of this study was to determine the range of adverse outcomes associated with chronic dizziness. DESIGN: Population-based prospective cohort study. SETTING AND PARTICIPANTS: A probability sample of 1087 persons, age 72 and older, living in the community. MEASUREMENTS: The following were measured: chronic dizziness, death, hospitalizations, falls, syncope, basic and instrumental activities of daily living, depressive symptoms, self-rated health, falls self-efficacy, and social activities. RESULTS: Of the 1087 participants, 261 (24%) reported chronic dizziness. Over 1 year of follow-up, chronic dizziness was not associated with mortality, hospitalization for any reason, or change in basic or instrumental activities of daily living, but was associated with risk of falling (unadjusted relative risk [RR] 1.35; 95% confidence interval [CI] 1.06-1.72) and with experiencing syncope (RR 2.31; 95% CI 1.24-4.30). After adjustment for baseline level, chronic dizziness also was associated with worsening of depressive symptoms, self-rated health, falls efficacy, and social activities. The relationship remained significant, after adjustment for potential confounding factors, for self-rated health (T-statistic -2.95, P = .003) and falls efficacy (T-statistic -2.68; P = .008), and was of marginal significance for depressive symptoms (T-statistic -1.73; P = .085). CONCLUSIONS: These results suggest that the goals of care for older persons with chronic dizziness should be redirected from solely identifying and treating discrete diseases--an often expensive and unrewarding task--toward reducing the symptoms of chronic dizziness and alleviating the resulting physical, psychological, and social disability. PMID- 10798470 TI - The association between parkinsonism, Alzheimer's disease, and mortality: a comprehensive approach. AB - BACKGROUND: The impact of parkinsonism on survival in older persons independent of dementia is not well understood. METHODS: Participants in the clinical examination of the Canadian Study of Health and Aging who had parkinsonism and were older than age 65 were identified. The impact of parkinsonism on 5-year survival was determined for a combined cohort with and without dementia, and a stratified analysis was then conducted for the subgroups with Alzheimer's disease (AD) and those without dementia. Subjects with a previous diagnosis of Parkinson's disease and those prescribed drugs causing extrapyramidal side effects were excluded. FINDINGS: A total of 721 subjects with AD and 1705 subjects without dementia were examined. After adjusting for age and residential status (community vs institution), parkinsonism was associated with poorer survival in the combined cohort (risk ratio 1.51; 95% CI, 1.22-1.85), in those with AD (risk ratio 1.34; 95% CI, 1.02-1.76), and those without dementia (risk ratio 1.54; 95% CI, 1.11-2.15). In the combined cohort, parkinsonism remained independently associated with higher mortality after adjusting for AD status (risk ratio 1.39; 95% CI, 1.13-1.72). In the subgroup with AD, parkinsonism remained associated with poorer survival after adjusting for severity of cognitive impairment (risk ratio 1.33; 95% CI. 1.04-1.74). INTERPRETATION: Parkinsonism is significantly associated with poorer survival in older persons, regardless of whether they have dementia. PMID- 10798471 TI - Fatigue in an older population. AB - OBJECTIVES: Fatigue is a common symptom that has not been studied well in the older populations. The purpose of this pilot study was to examine the epidemiology of fatigue symptoms in relation to demographic and medical characteristics of older patients in a long-term care setting. DESIGN: A cross sectional interviewer-assisted survey. PARTICIPANTS/SETTING: A total of 199 ambulatory older residents of a single residential care facility. MEASUREMENTS: Along with medical and demographic characteristics, the survey included mental status (Folstein), activities of daily living (Katz and Lawton), depression (Yesavage GDS), a 3-minute walk, a 7-item pain scale, and the modified Piper Fatigue Scale. RESULTS: One-hundred ninety-nine (65%) of 308 potential subjects completed the study (mean age 88 years, 82% female). Of these 199 subjects, 195 (98%) reported some fatigue symptoms (median duration 44 weeks). Significant (P < .0005) relationships were found between fatigue and GDS (r = 0.57), 3-minute walk (r = -0.29), Lawton IADLs (r = 0.31), pain (r = 0.36), and number of medications (r = 0.26). No significant relationships were found between fatigue and age, sex, Folstein score, or number of medical diagnoses. Multivariate regression analysis identified GDS, pain, number of medications and 3-minute walk as significant predictors of fatigue intensity (multiple R = 0.68, r2 = 0.46, P < .02). CONCLUSIONS: Fatigue is a symptom often found among older residents of a residential facility and has important implications for quality of life. Fatigue is poorly recognized and probably undertreated in older people. PMID- 10798472 TI - Mild cognitive impairment: potential pharmacological treatment options. AB - Both mild cognitive impairment and age-associated memory impairment are terms used to describe memory decline in otherwise healthy, intellectually intact individuals aged older than 50 years. It is estimated that up to 38% of the middle-aged and older population fulfill diagnostic criteria for this condition. Although the memory deficits observed in these individuals are fairly mild, they can interfere with day-to-day functioning. This article presents a review of the types of memory decline observed in older people, the diagnostic criteria used to define memory decline, the physiological and morphological brain changes that accompany aging, and the potential pharmacological treatment options, focusing on agents that have been evaluated in mildly cognitively impaired or normal older populations. PMID- 10798473 TI - Effects of an abilities-focused program of morning care on residents who have dementia and on caregivers. AB - OBJECTIVE: To conduct a controlled investigation to examine the effects of an abilities-focused program of morning care on the interaction behaviors and functioning of residents with dementia and on caregivers' interaction behaviors and perceptions of caregiving. DESIGN: A quasi-experimental, repeated measures design. SETTING: The study was conducted on four, nursing-home-level cognitive supports units in a geriatric care center. One of the units was randomly selected as the experimental unit; the other three served as controls. PARTICIPANTS: The final sample consisted of 40 residents (20 each in the experimental and three control groups) and 44 caregivers (16 on the experimental unit and 28 on the three control units). INTERVENTION: An educational program on delivering abilities-focused morning care, designed by the authors, was provided to caregivers on the experimental unit. MEASUREMENTS: Measures were taken at baseline and at 3 and 6 months postintervention with regard to residents' interaction behaviors, level of agitation, and level of function and to caregivers' interaction behaviors, perceived ease of caregiving, and level of stress. RESULTS: Repeated measures analysis of variance (RM-ANOVA) was used to compare the experimental and control groups in regard to changes in the outcomes over time. Results indicated that the abilities-focused program had statistically significant effects on (a) residents' personal attending and calm/functional behaviors, level of agitation, and levels of overall and social function, and (b) caregivers' verbal relevance and personal attending, relaxed, and social/flexible behaviors. CONCLUSIONS: The evidence suggests that both residents and caregivers benefit from morning care that is oriented toward the abilities of people with dementia. PMID- 10798474 TI - How the Balanced Budget Refinement Act of 1999 will affect geriatric medicine. PMID- 10798475 TI - The reciprocal relationship between disability and depression. PMID- 10798476 TI - Treating excess disability among cognitively impaired nursing home residents. PMID- 10798477 TI - Extracorporeal magnetic innervation (EXMI) therapy in the treatment of urinary incontinence in women: results from a single center. PMID- 10798478 TI - A case of lithium-induced asterixis. PMID- 10798479 TI - Medication adherence: is and why is older wiser? PMID- 10798480 TI - A GEM risk scoring protocol. PMID- 10798481 TI - Phospholipid fatty acid composition, vitamin E content and susceptibility to lipid peroxidation of duck spermatozoa. AB - Recent studies on chicken semen have suggested that the lipid and fatty acid composition of spermatozoa may be important determinants of fertility. Phospholipid fatty acid composition, vitamin E content and in vitro susceptibility to lipid peroxidation of duck spermatozoa were investigated using GC-MS and HPLC based methods. The total phospholipid fraction of duck spermatozoa was characterized by high proportions of the n-6 polyunsaturated fatty acids arachidonic (20:4n-6), docosatetraenoic (22:4n-6) and docosapentaenoic (22:5n-6) acids but a substantial proportion of the n-3 fatty acid docosahexaenoic (22:6n 3) acid was also present. Palmitic (16:0) and stearic (18:0) fatty acids were the major saturates in sperm phospholipids. Among the phospholipid classes, phosphatidylserine (PS) had the highest degree of unsaturation due to very high proportions of 22:6n-3, 22:5n-6, 22:4n-6 and 20:4n-6, comprising together more than 75% of total fatty acids in this fraction. Phosphatidylethanolamine (PE) also contained high proportions of these four C(20-22) polyunsaturates, which together formed 60% of total fatty acids in this phospholipid. Spermatozoa and seminal plasma of duck semen were characterized by unexpectedly low content of vitamin E, being more than 4-fold lower than in chicken semen. In duck semen the major proportion of the vitamin E (>70%) was located in the spermatozoa. The very high proportion of 22:6n-3 in PS and PE fractions of duck sperm lipids and the comparatively low levels of vitamin E could predispose semen to lipid peroxidation. Nevertheless the in vitro susceptibilities to Fe2+-stimulated lipid peroxidation of duck and chicken spermatozoa were very similar. The results of the study suggest that increased superoxide dismutase and glutathione peroxidase activity and increased antioxidant activity of seminal plasma may compensate for the low levels of vitamin E to help protect the membranes of duck spermatozoa, which exhibit a high degree of unsaturation from oxidative stress. PMID- 10798482 TI - Cryopreservation of bovine blastocysts obtained by intracytoplasmic sperm injection. AB - The freezability and survivability of zona-intact and zona-free (hatched) bovine blastocysts obtained by intracytoplasmic sperm injection (ICSI) were assessed. Day 7 or 8 blastocysts were cryopreserved by slow freezing using 1.5 M glycerol and 0.2 M sucrose. Embryos were exposed to solutions in a 2-step procedure at room temperature and frozen in a programmed cell freezer. Blastocysts that re expanded within 6 h of post-thaw culture were considered viable. The cleavage, morula and blastocyst development rates after ICSI were 52.4 (131/250), 39.7 (52/131), and 24.4% (32/131), respectively. Blastocyst stage embryos were randomly divided into 2 groups. The first group of embryos was frozen with their zonae intact, while the second group was allowed to hatch from their zonae during the additional 18 h culture, after which they were frozen. The data showed that more Group 2 blastocysts (14/16, 87.5%) than Group 1 (12/16; 75.0%; P<0.05) survived, and more zona-free bovine blastocysts frozen with glycerol as the cryoprotective agent (CPA) than zona-intact blastocysts after slow freezing retained their viability. PMID- 10798483 TI - Effect of trehalose and EDTA on cryoprotective action of ram semen diluents. AB - To obtain better ram semen extenders for artificial insemination (AI), we developed effective trehalose-containing hypertonic diluents. The cryoprotective action of trehalose has been explained by its dehydrating activity and interaction with cell membranes. Accordingly, we tested the cryopreserving capacity of different combinations of a Salamon's modified plus trehalose extender with EDTA. Evaluations were based on the percentage of motile spermatozoa and acrosome integrity, measured after thawing and after a 4-h post thaw resistance test at 37 degrees C. We conclude that the combination of trehalose plus EDTA confers the highest cryopreserving activity tested, not only for freeze-thawing but also for post-thawing resistance, possibly by removing calcium from the medium thereby preventing cation competition with trehalose for membrane-binding sites. PMID- 10798484 TI - Effect of superovulation induction on embryonic development on day 5 and subsequent development and survival after nonsurgical embryo transfer in pigs. AB - To evaluate the effects of eCG dosage on recovery and quality of Day 5 embryos and on subsequent development and survival after embryo transfer, batches of 5 to 10 donor sows were treated with 1000 or 1500 IU eCG. Recipients from the same batch were synchronously treated with 800 IU eCG. Ovulation was induced with 750 IU hCG (72 h after eCG) in donors and recipients. Donors were inseminated and embryos were collected at 162 h after hCG (120 h after ovulation). Ovulation rate was lower using 1000 IU eCG (28.5+/-11.7; n=48) than 1500 IU eCG (45.7+/-20.3; n=32; P<0.0001). Embryo recovery rate (82.9+/-16.9%) and percentage expanded blastocysts (56.2+/-31.4%) were similar (P>0.05). Expanded blastocysts from each group of sows were pooled into 2 groups within eCG treatment, containing embryos from normally ovulating sows (< or = 25 corpora lutea [CL]) or from superovulated sows (> 25 CL). Average diameter and number of cells of a random sample of the expanded blastocysts per pool were recorded. The average diameter of blastocysts (160.5+/-11.5 microm) was not affected by eCG dosage or ovulation rate (P>0.10). The average number of cells per embryo was higher in the 1000 IU eCG group (84.3+/-15.3) than in the 1500 IU eCG group (70.2+/-1.9; P<0.05) but was similar for normal and superovulated donors within each eCG group (P>0.10). Of the 4 groups, litters of 28 to 30 blastocysts were nonsurgically transferred to 27 synchronous recipients. Pregnant recipients were slaughtered on Day 37 after hCG treatment to evaluate embryonic development and survival. Pregnancy rate for the 1000 and 1500 IU eCG donor groups was 71% (10/14) and 46% (6/13; P>0.10), respectively. The number of implantations and fetuses for the 1000 IU eCG groups was 12.9+/-3.0 and 11.1+/-2.7, and 14.2+/-7.0 and 10.5+/-4.6, respectively, for the 1500 IU eCG groups (P>0.10). After post-priory categorizing the litters of blastocysts to below or above the average diameter (158 microm) of the transferred embryos, irrespective of eCG dosage or ovulation rate, the pregnancy rate was 43% (6/14) and 77% (10/13; P<0.10), respectively. Post-priory categorizing the transferred litters to below or above the average number of cells per embryo litter, irrespective of eCG dosage or ovulation rate, showed no differences in pregnancy rates or number of implantations and fetuses (P>0.10). It was concluded that eCG dosage affects embryonic development at Day 7 after hCG, and this effect was not due to ovulation rate. Embryonic survival after nonsurgical transfer was not related to eCG dosage but tended to be related to the diameter of the blastocysts. PMID- 10798485 TI - Long-term culture and characterization of goat primordial germ cells. AB - While the culture and identification of primordial germ cells (PGCs) in mice is established, only limited investigations on PGCs in livestock have been reported. This study was performed to characterize goat PGCs after culture and cryopreservation. Goat PGCs were isolated from Day 32 fetuses and cultured on a continuous cell line of murine embryonal fibroblasts (STO) as feeder-cells in the presence of leukemia inhibitory factor (LIF). The PGCs proliferated slowly and showed colony formation in early passages. Frozen-thawed PGCs continued to proliferate when stem cell factor (SCF) was added to the culture medium. However, differentiation into epithelial-like polygonal cells or neuronal cells was observed after 1 or 2 passages. The PGCs of 1 female and 1 male cell line were characterized by immunocytochemistry. The PGCs showed positive staining for anti stage-specific embryonic antigen-1 (SSEA-1) and FMA-1 (monoclonal antibody produced against a glycoprotein cell surface antigen of the embryonal carcinoma Nulli SCC1), whereas the reactivity to alkaline phosphatase (AP), an established marker for PGCs in mice, was inconsistent. After differentiation, PGCs lost their positive reaction to SSEA-1, EMA-1 and AP. In conclusion, SSEA-1 and EMA-1 can be used as reliable markers for identifying goat PGCs in addition to morphological criteria. The results indicate that goat PGCs can be kept in long-term culture without losing their morphological characteristics and their positive reaction to SSEA-1 and EMA-1, thus providing a promising source of donor-karyoplasts for nuclear transfer procedures. PMID- 10798486 TI - Exogeneous substances affecting development of in vitro-derived bovine embryos before and after embryonic genome activation. AB - Three experiments were conducted to evaluate how exogenous substances [fetal bovine serum (FBS), arachidonic acid (AA), glutathione (GSH), platelet-derived growth factor (PDGF), transforming growth factor-beta (TGF), fibroblast growth factor (FGF), insulin, transferrin and selenium (ITS)] affect preimplantation bovine embryo development. Cumulus-oocyte complexes (COC) were matured and fertilized in vitro, and their development was monitored up to 192 h post insemination in a two-step culture system. In Experiment 1, inseminated oocytes were cultured in modified bovine embryo culture medium (mBECM) supplemented with FBS or BSA for up to 60 h post insemination, and the resultant 8-cell embryos were then cultured singly in mBECM supplemented with AA+GSH+PDGF+TGF. More (P<0.005) blastocysts were derived from 8-cell embryos produced in media containing FBS than BSA. In Experiment 2, the 8-cell embryos produced in mBECM supplemented with FBS were cultured singly in mBECM as follows: 1) no supplementation; 2) AA and GSH or 3) AA, GSH, PDGF and TGF. Compared with no supplementation, a significant (P<0.05) increase in the proportion of 16-cell embryos and morulae was obtained after the addition of either AA+GSH or AA+GSH+PDGF+TGF. In Experiment 3, oocytes were cultured singly in mBECM as follows: 1) no supplementation; 2) AA+GSH+PDGF+TGF; 3) AA+GSH+PDGF+TGF and FGF; 4) AA+GSH+PDGF+TGF and ITS; 5) AA+GSH+PDGF+TGF, FGF and ITS or 6) FBS. Eight-cell embryos grown in each system were subsequently cultured singly in mBECM with AA+GSH+PDGF+TGF. More (P<0.05) 16-cell embryos were obtained in medium supplemented with either AA+GSH+PDGF+TGF and ITS or FBS than in unsupplemented medium. Fewer (P<0.05) oocytes developed to the 8-cell stage with the addition of AA+GSH+PDGF+TGF and FGF than without. In conclusion, embryo development to the blastocyst stage is regulated by exogenous AA, GSH, PDGF, FGF and ITS in a stage specific manner. PMID- 10798487 TI - Development of infantile rat ovaries autotransplanted after cryopreservation by vitrification. AB - We cryopreserved infantile rat ovaries by vitrification and assessed their viability by autotransplantation. Hemilateral ovarian transplantation was performed on rats on postnatal Days 10 to 12. The left ovary of each rat was dissected out, cryopreserved by vitrification using a modified vitrification solution (VS1), and then autotransplanted under the capsule of the right kidney. The right ovary of each rat was removed. For the control, the left ovary was dissected out from each rat and was immediately transplanted by the same procedure, without cryopreservation. Rats were nursed until weaning, and then the day of vaginal opening, estrous cyclicity from the day of vaginal opening until postnatal Day 84, and histology of ovarian grafts at postnatal Day 84 were examined. The time course of development of endocrine function of cryopreserved grafts was similar to that of fresh grafts. In ovarian transplants recovered on postnatal Day 84, antral follicles and corpora lutea (CL) were observed in addition to small follicles, although the number of antral follicles in cryopreserved grafts was smaller than in the fresh grafts. These results indicate that cryopreservation of ovarian tissue by vitrification can be used for the preservation of fertility and endocrine function of ovaries. PMID- 10798488 TI - Successful implantation of in vitro-matured, electro-activated oocytes in the pig. AB - In the present study, we derived parthenogenetic porcine fetuses from in vitro matured oocytes following a simple activation process in order to evaluate their developmental limitations in-vivo. Follicular oocytes collected from gilts at local slaughterhouses were matured for 48 h. They were subjected to a single square pulse of direct current for 100 microsec at 1,500 V/cm and then treated with 5 microg/mL cytochalasin B for 4 h to obtain activated diploid oocytes. The diploids were cultured in modified Whitten's medium until transfer. Diploids which had cleaved to the 2- and 3- to 4-cell stages were transferred to oviducts of recipients. Live and/or dead parthenogenetic fetuses were recovered in 6 of 8 trials at 17, 18, 19, 24 and 29 d post activation. The total proportion of fetuses to transferred diploids was 31.3% (62/198). When fetuses were recovered at 19 d post activation, the proportion of development into fetuses was 71% (15/21). Our results, however, suggest that periods of gestation longer than 19 d resulted in a decrease of these proportions to 45% (18/40) at 24 d and to 18% (7/40) at 29 d. The hearts were beating in nearly all of the fetuses recovered at 19, 24 and 29 d post activation. Thus, parthenogenetic porcine diploids developed to at least the stage of limb-bud formation beyond the early heart-beating stage. Abnormalities were also externally visible on some fetuses. Formation of cyst like structures in the heart and liver, and insufficient development of the head region and acephali were observed in some cases. PMID- 10798489 TI - Synchronization of ovulation in beef cows (Bos indicus) using GnRH, PGF2alpha and estradiol benzoate. AB - The objective of this study was to evaluate protocols for synchronizing ovulation in beef cattle. In Experiment 1, Nelore cows (Bos indicus) at random stages of the estrous cycle were assigned to 1 of the following treatments: Group GP controls (nonlactating, n=7) received GnRH agonist (Day 0) and PGF2alpha (Day 7); while Groups GPG (nonlactating, n=8) and GPG-L (lactating, n=9) cows were given GnRH (Day 0), PGF2alpha (Day 7) and GnRH again (Day 8, 30 h after PGF2alpha). A new follicular wave was observed 1.79+/-0.34 d after GnRH in 19/24 cows. After PGF2alpha, ovulation occurred in 19/24 cows (6/7 GP, 6/8 GPG, 7/9 GPG-L). Most cows (83.3%) exhibited a dominant follicle just before PGF2alpha, and 17/19 ovulatory follicles were from a new follicular wave. There was a more precise synchrony of ovulation (within 12 h) in cows that received a second dose of GnRH (GPG and GPG-L) than controls (GP, ovulation within 48 h; P<0.01). In Experiment 2, lactating Nelore cows with a visible corpus luteum (CL) by ultrasonography were allocated to 2 treatments: Group GPE (n=10) received GnRH agonist (Day 0), PGF2alpha (Day 7) and estradiol benzoate (EB; Day 8, 24 h after PGF2alpha); while Group EPE (n=11), received EB (Day 0), PGF2alpha (Day 9) and EB (Day 10, 24 h after PGF2alpha). Emergence of a new follicular wave was observed 1.6+/-0.31 d after GnRH (Group GPE). After EB injection (Day 8) ovulation was observed at 45.38+/-2.03 h in 7/10 cows within 12 h. In Group EPE the emergence of a new follicular wave was observed later (4.36+/-0.31 d) than in Group GEP (1.6+/-0.31 d; P<0.001). After the second EB injection (Day 10) ovulation was observed at 44.16+/-2.21 h within 12 (7/11 cows) or 18 h (8/11 cows). All 3 treatments were effective in synchronizing ovulation in beef cows. However, GPE and, particularly, EPE treatments offer a promising alternative to the GPG protocol in timed artificial insemination of beef cattle, due to the low cost of EB compared with GnRH agonists. PMID- 10798490 TI - D-penicillamine and granulosa cells can effectively extend the fertile life span of bovine frozen-thawed spermatozoa in vitro: effect on fertilization and polyspermy. AB - The effect of D-penicillamine on the fertile life span of frozen-thawed bull spermatozoa was studied in Experiment 1. After thawing, the washed spermatozoa were incubated for 8 h in fertilization medium with 1 mg/mL polyvinyl alcohol (PVA) and 0.5 mg/mL D-penicillamine. The addition of cumulus-free oocytes together with 10% of bovine serum (BOS) + 4 IU/mL heparin to the 8-h incubated spermatozoa resulted in high fertilization and polyspermy rates (97/103, 94.2% and 49/97, 50.5%, respectively). When BOS was substituted with 3 mg/mL of BSA, the fertilization and polyspermy rates decreased to 33/91 (36.3%) and 3/33 (9.1%), respectively. The total absence of fertilization was observed after substitution of proteins with PVA. The 8-h sperm incubation in fertilization medium without D-penicillamine and following fertilization in medium + different capacitation supplements resulted in the total absence of or a very low fertilization rate. In Experiment 2, the first set of cumulus-oocyte complexes (COC) and cumulus-free oocytes were fertilized for 8 h with nonincubated spermatozoa. The second set of COC and cumulus-free oocytes were fertilized in the same wells with spermatozoa after removal of the first set. The fertilization rate for the first set of COC and cumulus-free oocytes was 65/67 (97%) and 73/73 (100%), respectively, with 21/65 (32.3%) and 32/73 (43.8%) polyspermy, respectively. In the second set, the high penetration rate (67/73, 91.8%) was observed only for COC, while that for cumulus-free oocytes (19/76, 25%) was significantly lower (P < 0.01). In Experiment 3, the fertilization rate of COC in medium + PVA + 10 or 100 IU/mL heparin was 69/92 (75%) and 52/74 (70.2%), respectively, with no polyspermy. In medium + BSA + 10 or 100 IU/mL heparin, the fertilization rate of COC was 64/72 (88.9%) and 70/79 (88.6%), respectively, with polyspermy at 2/64 (3.1%) and 7/70 (10%), respectively. In medium + BOS + 10 or 100 IU/mL heparin, the fertilization rate was 79/82 (96.3%) and 79/80 (98.7%), respectively, with a significantly (P < 0.05) higher polyspermy rate (34/79, 43% and 30/79, 38%, respectively). The vitality and capacitation changes of frozen thawed and 8-h incubated spermatozoa were assessed by Hoechst and chlortetracycline (CTC) staining. PMID- 10798491 TI - Physiological basis for use of insulin-like growth factors in reproductive applications: a review. AB - The insulin-like growth factors (IGF-I and -II) are ubiquitously expressed factors that regulate cell growth, differentiation and maintenance of differentiated cell function. All aspects of male and female reproduction are influenced by the IGF system. This review will examine the IGF system as it pertains to reproductive physiology and applications. PMID- 10798492 TI - The protective action of polyvinyl alcohol during rapid-freezing of mouse embryos. AB - Biological products like serum and BSA are routinely used in embryo freezing solutions. These products are undefined and can potentially expose the embryos to infectious agents. Therefore, this experiment was designed to evaluate in vitro and in vivo survival of mouse embryos frozen in solutions supplemented with a chemically defined macromolecule, polyvinyl alcohol (PVA). Morula-stage embryos from superovulated mice were collected, frozen by a rapid freezing procedure, and cryoprotectant diluted out (after thawing) in media supplemented with either 10% fetal calf serum (FCS), 0.1 mg/mL PVA, or a combination of 10% FCS and 0.1 mg/mL PVA. Frozen-thawed (good to excellent quality) and nonfrozen (control, collected in FCS supplemented medium) embryos were cultured in medium M16 (32) supplemented with either 4 mg/mL BSA or 0.1 mg/mL PVA for 72 h. Embryos frozen in solutions supplemented with FCS or PVA and nonfrozen embryos were transferred to pseudopregnant recipients. Recipients were humanly killed on Day 15 after transfer, and the rate of implantation and percentage of live fetuses were recorded. The supplementation of collection, freezing and cryoprotectant dilution solutions with FCS, PVA or FCS plus PVA did not influence (P > 0.05) the rate of survival and in vitro development of embryos to hatched/hatching blastocyst stage. However, a higher (P < 0.01) in vitro development rate to hatching/hatched stage was recorded when frozen-thawed embryos were cultured in medium supplemented with BSA than with PVA. There was no difference (P > 0.05) in the rate of implantation (68 vs 72%) or percentage of live fetuses (62 vs 60%) between pregnant recipients with embryos frozen in medium with FCS or PVA. The rate of implantation and development of embryos frozen in medium supplemented with PVA or FCS was comparable (P > 0.05) to that of nonfrozen embryos. It may be concluded that PVA can be substituted for FCS in medium for freezing mouse embryos; however, it can not be completely substituted for BSA in the in vitro culture of embryos to the hatched blastocyst stage. PMID- 10798493 TI - Field investigations of bacterial contaminants and their effects on extended porcine semen. AB - Field investigations (n=23) were made over a 3-yr period at North American boar studs and farms in which the primary complaint was sperm agglutination in association with decreased sperm longevity of extended semen, and increased regular returns to estrus and/or vaginal discharges across parity. Microscopic examination of extended semen from these units revealed depressed gross motility (usually <30%), sperm agglutination, and sperm cell death occurring within 2 d of semen collection and processing regardless of the semen extender used. The extended semen exhibited a high number of induced acrosome abnormalities (>20%). Sample pH was acidic (5.7 to 6.4) in 93% of the submitted samples. Aerobic culture yielded a variety of bacteria from different genera. A single bacterial contaminant was obtained from 66% of the submitted samples (n=37 doses); 34% contained 2 or more different bacterial genera. The most frequently isolated contaminant bacteria from porcine extended semen were Alcaligenes xylosoxydans (n=3), Burkholderia cepacia (n=6), Enterobacter cloacae (n=6), Escherichia coli (n=6), Serratia marcescens (n=5), and Stenotrophomonas [Xanthomonas] maltophilia (n=6); these 6 bacteria accounted for 71% of all contaminated samples, and were spermicidal when re-inoculated and incubated in fresh, high quality extended semen. All contaminant bacteria were found to be resistant to the aminoglycoside gentamicin, a common preservative antibiotic used in commercial porcine semen extenders. Eleven genera were spermicidal in conjunction with an acidic environment, while 2 strains (E. coli, S. maltophilia) were spermicidal without this characteristic acidic environment. Bacteria originated from multiple sources at the stud/farm, and were of animal and nonanimal origin. A minimum contamination technique (MCT) protocol was developed to standardize hygiene and sanitation. This protocol focused on MCT's during boar preparation, semen collection, semen processing and laboratory sanitation. Implementation of the MCT, in addition to specific recommendations in stud management, resulted in the control of bacterial contamination in the extended semen. PMID- 10798494 TI - The decline of porcine sperm motility by geldanamycin, a specific inhibitor of heat-shock protein 90 (HSP90). AB - Sperm motility is an important parameter for fertility. The molecular mechanisms of mammalian sperm motility are still largely undefined. Our previous observations suggested that heat shock protein 90 (HSP90) may be associated with porcine sperm motility. The aim of the present study was to further characterize the plausible novel function of HSP90 on sperm motility. Semen from normal, sexually mature boars with sperm motility higher than 80% was used. An HSP90 specific inhibitor, geldanamycin (GA), was added to diluted semen at 0.5, 1.0, 2.5 or 5.0 microg/mL and the semen was then incubated at 37 degrees C for 15, 30, 45 or 60 min. Sperm motility was determined by using computer-assisted semen analyzer at the end of incubation. The results indicated that GA significantly reduced sperm motility in a dose and time dependent manner. Moreover, incubation of semen with 5.0 microg/mL GA for 15 min completely stopped sperm motility. To test the reversibility of the GA effect on sperm motility, GA was removed after 30 min incubation and was replaced with fresh extender alone or with extender plus 5 mM caffeine, then incubated for another 15, 30, 45 or 60 min. The results showed that simply removing GA did not reverse the inhibitory effect on sperm motility, while adding caffeine partially reversed this inhibitory effect. However, the effect of 2.5 or 5.0 microg/mL GA was not reversed by caffeine. Considering the specificity of GA targeting to HSP90, the above observations suggested that HSP90 may play a crucial role in regulating porcine sperm motility. PMID- 10798495 TI - Semen quality during vincristine treatment in dogs with transmissible venereal tumor. AB - The aim of this study was to evaluate the direct effects of vincristine on semen quality in dogs with transmissible venereal tumor (TVT). We examined the semen of 17 dogs suffering from TVT during vincristine treatment. Each animal received 0.6 mg, i.v. vincristine sulphate per square meter of body surface, per week for 4 wk until complete regression of the tumor. The following semen parameters were evaluated: semen volume (second fraction), sperm concentration, total spermatozoa per ejaculate, percentage of progressively motile spermatozoa, percentage of dead spermatozoa, percentage of swollen spermatozoa (hypo-osmotic swelling test) and percentage of morphologically abnormal spermatozoa (primary and secondary defects). Semen was collected and evaluated prior to the beginning of treatment, 3 d after each vincristine injection and 15 d after the last injection. Semen characteristics transiently deteriorated during treatment, but returned to normal 15 d later. These changes were attributed to a direct effect of vincristine on the extragonadal spermatozoal reserves contained in the epididymis and ductus deferens. A GnRH stimulation test was also performed after each semen collection in order to assess the function of the hypothalamic-pituitary-Leydig cell axis. No effect was noted on the above axis. PMID- 10798496 TI - Characterization of gossypol-induced sperm abnormalities in bulls. AB - To characterize sperm abnormalities induced by gossypol in cattle, young Brahman bulls (n=8) received either cottonseed meal delivering 8.2 g free gossypol/bull/d (treatment, n=4) or isonitrogenous soybean meal (control, n=4) for 11 wk. At slaughter, semen was collected from the following extragonadal sites: mediastinum/rete testis (Site 1), caput (Site 2), corpus (Site 3) and cauda epididymis (Site 4), and ductus deferens (Site 5). At least 200 fixed spermatozoa per site were examined via differential-interference-phase contrast (DIC) microscopy, with electron microscopy (EM) being employed with select samples. Sperm midpiece abnormalities were categorized as aplastic, fragile or asymmetric, with detached sperm heads being tabulated separately. Of these, aplastic defects were considered most likely to occur during spermatogenesis. Bull sperm midpiece lesions induced by gossypol were ultrastructurally similar to those observed in other, nonruminant, species. Combined midpiece abnormalities generally increased with extragonadal passage (EGP) in the treated bulls, as did fragile and asymmetric but not aplastic midpieces, or detached sperm heads. This pattern of EGP changes in bull sperm morphology following gossypol spermatoxicity suggests that structural weakness induced during spermatogenesis leads to secondary spermatozoal changes during EGP, possibly due to the imposition of motility stressors upon prior weakened structures. PMID- 10798497 TI - Simulation studies of vestibular macular afferent-discharge patterns using a new, quasi-3-D finite volume method. AB - A quasi-three-dimensional finite-volume numerical simulator was developed to study passive voltage spread in vestibular macular afferents. The method, borrowed from computational fluid dynamics, discretizes events transpiring in small volumes over time. The afferent simulated had three calyces with processes. The number of processes and synapses, and direction and timing of synapse activation, were varied. Simultaneous synapse activation resulted in shortest latency, while directional activation (proximal to distal and distal to proximal) yielded most regular discharges. Color-coded visualizations showed that the simulator discretized events and demonstrated that discharge produced a distal spread of voltage from the spike initiator into the ending. The simulations indicate that directional input, morphology, and timing of synapse activation can affect discharge properties, as must also distal spread of voltage from the spike initiator. The finite volume method has generality and can be applied to more complex neurons to explore discrete synaptic effects in four dimensions. PMID- 10798498 TI - A population density approach that facilitates large-scale modeling of neural networks: analysis and an application to orientation tuning. AB - We explore a computationally efficient method of simulating realistic networks of neurons introduced by Knight, Manin, and Sirovich (1996) in which integrate-and fire neurons are grouped into large populations of similar neurons. For each population, we form a probability density that represents the distribution of neurons over all possible states. The populations are coupled via stochastic synapses in which the conductance of a neuron is modulated according to the firing rates of its presynaptic populations. The evolution equation for each of these probability densities is a partial differential-integral equation, which we solve numerically. Results obtained for several example networks are tested against conventional computations for groups of individual neurons. We apply this approach to modeling orientation tuning in the visual cortex. Our population density model is based on the recurrent feedback model of a hypercolumn in cat visual cortex of Somers et al. (1995). We simulate the response to oriented flashed bars. As in the Somers model, a weak orientation bias provided by feed forward lateral geniculate input is transformed by intracortical circuitry into sharper orientation tuning that is independent of stimulus contrast. The population density approach appears to be a viable method for simulating large neural networks. Its computational efficiency overcomes some of the restrictions imposed by computation time in individual neuron simulations, allowing one to build more complex networks and to explore parameter space more easily. The method produces smooth rate functions with one pass of the stimulus and does not require signal averaging. At the same time, this model captures the dynamics of single-neuron activity that are missed in simple firing-rate models. PMID- 10798499 TI - On the simulation of large populations of neurons. AB - The dynamics of large populations of interacting neurons is investigated. Redundancy present in subpopulations of cortical networks is exploited through the introduction of a probabilistic description. A derivation of the kinetic equations for such subpopulations, under general transmembrane dynamics, is presented. The particular case of integrate-and-fire membrane dynamics is considered in detail. A variety of direct simulations of neuronal populations, under varying conditions and with as many as O(10(5)) neurons, is reported. Comparison is made with analogous kinetic equations under the same conditions. Excellent agreement, down to fine detail, is obtained. It is emphasized that no free parameters enter in the comparisons that are made. PMID- 10798501 TI - Pediatric iron poisonings in the United States. AB - BACKGROUND: Iron overdose is considered a leading cause of poisoning-related injury and death in young children. This report analyzes the nature, trend, and hazard patterns of unintentional pediatric iron overdoses in the United States from 1980 to 1996. METHODS: Analyses include multiple regression and correlation analysis of national data on pediatric iron ingestion-related injuries and deaths and review of in-depth investigation case reports. Data sources include files of the US Consumer Product Safety Commission, National Center for Health Statistics, American Association of Poison Control Centers, and US Census Bureau. RESULTS: Pediatric iron-related injuries increased 150% in 1986, from an annual average of 1,200 from 1980 through 1985 to 3,000 from 1986 through 1996. No such annual trend occurred before or after 1986. About one third of the injuries from 1980 through 1996 involved infants under 2 years old, a third involved 2-year-olds, and a third involved children 3 or 4 years old. Pediatric iron-related fatalities increased in 1986, peaked at 10 in 1991, and declined to 2 by 1995. The children often obtained the iron from a child-resistant container opened by themselves or another child or left open or improperly closed by an adult. CONCLUSIONS: Iron overdose remains a significant public health threat to young children. The frequency of pediatric iron overdose injuries increased in 1986 and has not declined. Unit-dose packaging of potent iron supplements is expected to reduce the frequency of severe pediatric iron overdose incidents. PMID- 10798502 TI - Radioguided surgical advancements for head and neck oncology. AB - Radioguided surgery is an innovative means by which a radionuclide is used to preoperatively image and intraoperatively visualize a structure of interest to the surgeon for excisional biopsy. This technology has allowed a cost-effective, highly specific means by which to locate a structure (usually a lymph node) and access it for pathologic analysis. The result of radioguided surgery is increased specificity in tissue obtained for biopsy, minimal access incisions, and the reduction of inpatient hospital utilization. Radioguided surgery should not be confused with radiosurgery, which is the stereotactic application of external beam radiation, usually for intracranial tumors. PMID- 10798500 TI - Models of calmodulin trapping and CaM kinase II activation in a dendritic spine. AB - Activation of calcium/calmodulin-dependent protein kinase II (CaMKII) by calmodulin following calcium entry into the cell is important for long-term potentiation (LTP). Here a model of calmodulin binding and trapping by CaMKII in a dendritic spine was used to estimate levels and durations of CaMKII activation following LTP-inducing tetani. The calcium signal was calcium influx through NMDA receptor channels computed in a highly detailed dentate granule cell model. Calcium could bind to calmodulin and calmodulin to CaMKII. CaMKII subunits were either free, bound with calmodulin, trapped, autonomous, or capped. Strong low frequency tetanic input produced little calmodulin trapping or CaMKII activation. Strong high-frequency tetanic input caused large numbers of CaMKII subunits to become trapped, and CaMKII was strongly activated. Calmodulin trapping and CaMKII activation were highly dependent on tetanus frequency (particularly between 10 and 100 Hz) and were highly sensitive to relatively small changes in the calcium signal. Repetition of a short high-frequency tetanus was necessary to achieve high levels of CaMKII activation. Three stages of CaMKII activation were found in the model: a short, highly activated stage; an intermediate, moderately active stage; and a long-lasting third stage, whose duration depended on dephosphorylation rates but whose decay rate was faster at low CaMKII activation levels than at high levels. It is not clear which of these three stages is most important for LTP. PMID- 10798503 TI - Persistence of cerebral blood flow after brain death. AB - Persistent cerebral blood flow occasionally confounds confirmatory tests for brain death and results in the anguish of delayed diagnosis, unnecessary use of expensive resources, and loss of transplant opportunities. We reviewed the literature to examine the reasons, frequency, and meaning of this problem. We found that this phenomenon occurs: (1) before increasing intracranial pressure completely shuts down flow; (2) in infants with pliable skulls; and with (3) decompressing fractures, (4) ventricular shunts, (5) ineffective deep brain flow, (6) reperfusion, (7) brain herniation, (8) jugular reflux, (9) emissary veins, and (10) pressure injection artifacts. Isolated venous sinus visualization is common (occurring in up to 57%) but represents trivial blood flow and confirms brain death. Arterial flow is much less common (2.6% incidence in our series). Normal flow occurs but is rare. Arterial flow does not exclude brain death, but the diagnosis should be confirmed by repeated studies or other means. PMID- 10798504 TI - Hormesis and health: a little of what you fancy may be good for you. AB - The term hormesis refers to beneficial effects from low doses of potentially harmful substances. Although there are many laboratory examples of this phenomenon, it remains controversial and has never become widely accepted by the health community. This review goes beyond the laboratory and describes many clinical and common sense, real-world examples of hormesis that often go unrecognized. Many vitamins and minerals are essential for life at low doses but toxic at higher ones. Similarly, exercise, caloric restriction, and alcohol consumption are examples of processes that are harmful in the extreme but beneficial in moderation. This review also highlights possible reasons why acceptance of the hormetic paradigm has lagged. These include high-dose toxicologic testing that precludes the demonstration of low-level effects and the threat posed by hormesis to the currently accepted precautionary principle, which assumes that any dose of a chemical is potentially harmful. PMID- 10798505 TI - Population-based survey of complementary and alternative medicine usage, patient satisfaction, and physician involvement. South Carolina Complementary Medicine Program Baseline Research Team. AB - BACKGROUND: With an increasing proportion of Americans using complementary or alternative medicine (CAM), physicians need to know which patients are using CAM to effectively manage care. METHODS: In this cross-sectional study, telephone interviews were conducted with 1,584 South Carolina adults (ages 18 and older); 66% responded to the survey of demographics, general health, frequency of CAM use, perceived CAM effectiveness, and physician knowledge of CAM use. RESULTS: A total of 44% had used a CAM during the past year. Increasing age and higher education were significantly associated with CAM use. More than 60% perceived CAM therapy as very effective, and 89% said they would recommend CAM to others. Physicians were unaware of CAM use in 57% of their patients using CAM. CONCLUSION: Complementary or alternative medicine use in this rural Southern state is similar to national usage. Users view CAM as effective. Physicians are frequently unaware of patients' CAM use. More research is needed to establish CAM effectiveness and how CAM affects medical care, training, and public health. PMID- 10798506 TI - Blunt pancreatic trauma: a difficult injury. AB - The diagnosis and initial management of hemodynamically stable blunt pancreatic trauma should include a high index of suspicion when a patient has sustained a severe force vector in the anterior-posterior direction of the abdominal cavity. Patients who are hemodynamically stable should have physical examination, computed tomography (CT) of the abdomen, and serial determinations of serum amylase levels. If CT shows any suggestion of injury to the pancreas, such as fluid between the splenic vein and the posterior border of the pancreas, follow up endoscopic retrograde pancreatography (ERP) should be done. In patients with a significant retroperitoneal hematoma, ERP should be followed by noncontrast CT for the evaluation of extravasation of contrast material. If ERP shows ductal disruption or follow-up CT shows extravasation of contrast, operative management is indicated. The astute physician should always remember that the most difficult aspect of pancreatic injury is timely diagnosis. PMID- 10798507 TI - Income inequality and all-cause mortality in the 100 counties of North Carolina. AB - BACKGROUND: This study evaluated the relationship between income inequality and all-cause mortality in the 100 counties of North Carolina. METHODS: Mortality data for 1985 to 1994 came from the National Center for Health Statistics and household income data from the 1990 Census. Associations between county-level income inequality and age-adjusted, all-cause mortality rates were evaluated in stratified and regression analyses. RESULTS: Stratified analyses suggest that all cause mortality was directly related to income inequality in all 100 counties and in non-metropolitan statistical area (MSA) counties. This relationship was statistically significant, controlling for per capita income in regression analyses for all age groups except > or =65 years among all 100 counties and for all ages combined and for ages 35 to 64 among non-MSA counties. CONCLUSIONS: A relationship between income inequality and all-cause mortality previously identified nationally among states and MSAs was also found among all counties and non-MSA counties of North Carolina in 1985 to 1994. PMID- 10798508 TI - Liver transplantation in the era of cost constraints. AB - BACKGROUND: The issue of containing cost has had a significant impact on organ transplantation. After our institution's 500th liver transplant, we critically examined the impact of the changing health care environment on liver transplantation. METHODS: We retrospectively analyzed 500 consecutive liver transplants done in the period of 1989 to 1998. RESULTS: Comparing the first 100 liver transplants to the last 100, patient demographics did not change significantly; however, mean waiting times increased significantly, from 30.4 days to 146.7 days, and median hospital stay decreased from 20.2 days to 10.9 days. One-year patient and graft survivals were not significantly different, 93.6% versus 96.5% and 88.0% versus 95.7%, respectively. CONCLUSIONS: Despite transplants in patients at higher risk and discharging patients sooner after transplantation, surgical results and patient survivals remained excellent. This was accomplished through improvements and modification of immunosuppression, outpatient treatment of uncomplicated acute rejection, and emphasis on close outpatient follow-up. PMID- 10798509 TI - Limited utility of routine drug screening in trauma patients. AB - BACKGROUND: Routine toxicology screening of seriously injured patients has become the standard of care in most trauma centers. However, the benefit of drug screening in acute trauma is unproven. We reviewed the impact of positive drug screening results on patient care within the first 3 days of treatment. METHODS: We retrospectively reviewed the charts of seriously injured patients admitted to an American College of Surgeons-certified level I trauma center over a 5-year period. Modifications of therapeutic regimens based on positive toxicology results were noted. Using current financial data, charges for toxicology were calculated. RESULTS: Between January 1, 1990, and December 31, 1995, 2,678 trauma patients had drug screening. Of these, 414 (15%) had detectable quantities of the following intoxicants: opiates, barbiturates, amphetamines, phencyclidine hydrochloride (PCP), cocaine, marijuana, or benzodiazepines. Review of all 401 available charts failed to identify any cases in which treatment was altered by a positive toxicology result. Hospital costs related to routine screening were $138,587, while charges to patients amounted to $538,278. CONCLUSIONS: Routine toxicology does not alter or improve the immediate care of the injured patient. Routine drug screening is expensive, and benefits were not easily documented. The policy of routine toxicology screening in trauma centers should be reevaluated. PMID- 10798510 TI - Clinical and biopsy specimen features in black and white men with clinically localized prostate cancer. AB - BACKGROUND: The incidence of prostate cancer in black men is estimated to be 30% to 50% higher than among age-matched white men, and black men have a twofold higher mortality rate. To determine whether racial differences exist in men with similarly staged disease, we compared clinical and pathologic features in black and white men with clinical stage T1-T2 prostate cancer. METHODS: We retrospectively reviewed the records of all men who had prostate biopsy at our facilities. Men were included in this study if they were of black or white race, if the clinical stage of their prostate cancer was T1-T2, and if detailed information regarding their prostate biopsy was available. RESULTS: Black men were found to have significantly higher serum prostate-specific antigen (PSA) levels, but no other differences were noted in the other clinical or pathologic features studied. CONCLUSION: While racial differences in the incidence and mortality of prostate cancer are well known, differences in the clinical and pathologic features between black and white men with similarly staged disease have been poorly studied. Our results suggest that serum PSA levels are higher in black men, but no other differences were noted. This suggests that early detection programs for prostate cancer in black men can potentially decrease prostate cancer mortality. PMID- 10798511 TI - Blunt trauma in intoxicated patients: is computed tomography of the abdomen always necessary? AB - BACKGROUND: Physical examination to detect abdominal injuries has been considered unreliable in alcohol-intoxicated trauma patients. Computed tomography (CT) plays the primary role in these abdominal evaluations. METHODS: We reviewed medical records of all blunt trauma patients admitted to our trauma service from January 1, 1992, to March 31, 1998. Study patients had a blood alcohol level > or =80 mg/dL, Glasgow Coma Scale (GCS) score of 15, and unremarkable abdominal examination. RESULTS: Of 324 patients studied, 317 (98%) had CT scans negative for abdominal injury. Abdominal injuries were identified in 7 patients (2%), with only 2 (0.6%) requiring abdominal exploration. A significant association was found between major chest injury and abdominal injury. CONCLUSION: The incidence of abdominal injury in intoxicated, hemodynamically stable, blunt trauma patients with a normal abdominal examination and normal mentation is low. Physical examination and attention to clinical risk factors allow accurate abdominal evaluation without CT. PMID- 10798512 TI - Measuring self-efficacy in substance abuse intervention in obstetric practices. AB - BACKGROUND: This study presents findings on the effect of an office-based obstetrics training program (given the acronym OBIWOM) on substance use assessment, management, and referral self-efficacy among obstetricians and staff of private, community-based obstetric practices. METHODS: Participants were obstetricians and staff from 10 of 27 available community-based, private obstetric practices in the target areas, for a practice participation rate of 37%. This study used a delayed treatment design to compare self-efficacy between practice staff randomly assigned to an immediate or delayed intervention group. RESULTS: Self-efficacy increased significantly after intervention from baseline to first follow-up for the immediate group, while no change was shown for the delayed (control) group. The impact of the intervention on self-efficacy was replicated in the assessment construct only after the intervention for the delayed group. CONCLUSION: This research shows that education and training can effectively improve self-efficacy in obstetricians and their staff in the management of substance use and pregnancy. PMID- 10798513 TI - Left main coronary artery rotational atherectomy and stenting. AB - BACKGROUND: Coronary artery bypass surgery is a difficult option in patients who are not candidates for bypass surgery and high-risk patients with critical left main coronary artery (LMCA) disease. We report outcomes and short-term follow-up of patients who had LMCA rotational atherectomy and/or stenting, assess the role of these interventions in protected and unprotected significant LMCA stenosis, and review the literature. METHODS: We reviewed the cases of seven men with critical LMCA stenosis for whom coronary artery bypass surgery was considered a high risk. Five patients had rotational atherectomy, one had coronary artery stenting, and one had both. RESULTS: In all cases, angiographic success was achieved, and symptoms were relieved. Six patients were discharged from the hospital in 3 to 6 days. One patient who had cardiogenic shock, respiratory failure, and acute renal failure before the procedure died of arrhythmia 4 days afterward. Another patient had elective coronary artery bypass graft surgery 3 weeks later for recurrent angina. Cardiac catheterization was repeated in 1 month for chest pain in three patients at 4 to 7 months follow-up, and none had progression of residual stenosis in the LMCA. CONCLUSIONS: Our study suggests that LMCA rotational atherectomy and stenting are safe and effective revascularization procedures in high-risk patients and patients who are not candidates for bypass surgery. PMID- 10798514 TI - Frequency of lipid-lowering drug therapy after coronary and carotid operations for atherosclerosis. AB - BACKGROUND: Coronary artery bypass grafting (CABG) and carotid endarterectomy (CE) are palliative operations for patients with severe atherosclerosis of the coronary or carotid arteries. Secondary preventive measures after these operations have received little attention. METHODS: We conducted a retrospective observational study to determine the frequency of lipid-lowering drug therapy before and after CABG or CE done in 1997. RESULTS: Of the 204 patients who had CABG, 56 (27%) were receiving lipid-lowering drug therapy before admission, and 71 (37%) of 192 were discharged receiving it. Of the 59 patients who had CE, only 12 (20%) were taking a lipid-lowering drug before admission, and 8 (14%) upon discharge. Of those 59 patients, 23 (39%) had coronary artery disease. CONCLUSION: Secondary prevention after coronary or carotid operations for atherosclerosis has been underemphasized at our institution, prompting the establishment of a postoperative risk reduction program. PMID- 10798515 TI - Medical complications due to mothball abuse. AB - Inhalant abuse is a major public health problem that has been associated with numerous acute and chronic medical problems. Inhalants are defined as volatile organic substances and are found in common household and commercial products that are easily accessible, inexpensive, and legally obtained. We discuss a rare case of mothball abuse predominantly by inhalation. The patient had acute peripheral neuropathy and chronic renal failure, both of which were concluded to be casually related to mothball abuse. We briefly review the abuse potential and medical complications of naphthalene and paradichlorobenzene, the compounds in mothballs. Our case illustrates that common household products not usually identified as recreational drugs can be abused; furthermore, linking specific medical problems with particular inhalants can be difficult because of confounding variables. PMID- 10798516 TI - Open air carbon monoxide poisoning in a child swimming behind a boat. AB - Carbon monoxide (CO) poisoning is the most common fatal poisoning in the United States. The circumstances often involve an unsuspected increase of CO in an enclosed environment. Victims often are unaware that their activity or environment placed them at risk for CO poisoning. The possibility of open air CO poisonings was first reported in 1987. We present a case of open air CO poisoning resulting in neurologic depression and a markedly elevated carboxyhemoglobin level in a child who had been swimming behind a house boat. Emergency physicians and pediatricians should be aware of the possibility of accidental open air CO poisoning in children and adults who swim around recreational boats. PMID- 10798517 TI - Tick paralysis syndrome in a 5-year-old girl. AB - Tick paralysis syndrome (TPS) is an uncommon cause of ascending paralysis in children. Familiarity with its clinical features is important, since prompt diagnosis and removal of the tick is curative. We report the case of a 5-year-old girl with TPS manifested as lower extremity ataxia and paralysis and briefly discuss the salient features of TPS. PMID- 10798518 TI - Pulmonary thromboembolism during transurethral resection of the prostate. AB - Although perioperative pulmonary thromboembolism (PTE) is a common complication of surgery, intraoperative pulmonary thromboembolism is unusual. We report the occurrence of PTE during transurethral resection of the prostate. We believe this to be the first report of such a case. PMID- 10798519 TI - Melanoma vaccines as a therapeutic option. PMID- 10798520 TI - Sinus bradycardia due to mitoxantrone. PMID- 10798521 TI - Social events and scientific innovations may affect the content of delusions. PMID- 10798522 TI - Benzodiazepine-induced hyperphagia: development and assessment of a 3D pharmacophore by computational methods. AB - Benzodiazepine receptor (BDZR) ligands are structurally diverse compounds that bind to specific binding sites on GABA(A) receptors and allosterically modulate the effect of GABA on chloride ion flux. The binding of BDZR ligands to this receptor system results in activity at multiple behavioral endpoints, including anxiolytic, sedative, anticonvulsant, and hyperphagic effects. In the work presented here, a computational procedure developed in our laboratory has been used to obtain a 3D pharmacophore for ligand recognition of the GABA(A)/BDZRs initiating the hyperphagic response. To accomplish this goal, 17 structurally diverse compounds, previously assessed in our laboratory for activity at the hyperphagic endpoint, were used. The result is a four-component 3D pharmacophore. It consists of two proton acceptor atoms, the centroid of an aromatic ring and the centroid of a hydrophobic moiety in a common geometric arrangement in all compounds with activity at this endpoint. This 3D pharmacophore was then assessed and successfully validated using three different tests. First, two BDZR ligands, which were included as negative controls in the set of seventeen compounds used for the pharmacophore development, did not fit the pharmacophore. Second, some benzodiazepine ligands known to have activity at the hyperphagia endpoint, but not included in the pharmacophore development, were used as positive controls and were found to fit the pharmacophore. Finally, using the 3D pharmacophore developed in the present work to search 3D databases, over 50 classical benzodiazepines were found. Among them, were benzodiazepine ligands known to have an effect at the hyperphagic endpoint. In addition, the novel compounds also found in this search are promising therapeutic agents that could beneficially affect feeding behavior. PMID- 10798523 TI - Computational analysis of the first biheterocyclization site of the antibiotic microcin B17. AB - Microcin B 17 (MccB17) undergoes an enzyme catalyzed posttranslational modification to form four oxazole and four thiazole rings. Four of these rings form 4,2 - connected biheterocyclic functionalities. In this study, the hexapeptide sequence surrounding the first biheterocyclization site of microcin B17 was examined using computational calculations and database analysis to see if it was preorganized for cyclization in a manner similar to that found in the autocatalytic posttranslational cyclization of Green Fluorescent Protein (GFP). Attention was focused on the intermolecular distances between the sulfur and oxygen atoms of the cysteine and serine residues and the carbonyl carbons which they attack in the ring formation. Conformational searches located some low energy conformations that contained relatively short oxygen to carbonyl carbon distances, which indicated that the oxazole forming fragment in microcin B17 is preorganized for cyclization. However, the lack of any clear patterns for the sulfur to carbon distances show that the side-chain of cysteine does not adopt any low energy conformations that are geometrically preorganized for cyclization. The MccB17 synthetase enzyme complex which catalyzes the cyclization process therefore has both steric and electronic functions. The data obtained in this investigation is in agreement with empirical data which shows that biheterocyclization will only occur if the thiazole forms before the oxazole. PMID- 10798524 TI - Molecular mechanics evaluation of the proposed mechanisms for the degradation of urea by urease. AB - A thorough conformational search of all the conformations available to oxygen bound urea within wild-type urease was carried out. Identical low energy urea conformations were obtained by a Ramachandran type plot for the NHis272-Ni1-O Curea, and Ni1-O-Curea-Nurea dihedral angles. Ramachandran plots, with active sites and protonation states modified to model the different urease mechanisms, were used to evaluate the different mechanisms. Based upon the low energy conformations available to urea in the active site of wild-type urease one can conclude that the traditional "His320 acts as a base" mechanism is unlikely. while the N,O urea bridged and the reverse protonation mechanisms cannot be ruled out. A consensus hydrogen-bonding network that does not favor any of the mechanisms has been reconfirmed by the extensive conformational search. PMID- 10798525 TI - A new approach for the calculation of the energy of van der Waals interactions in macromolecules of globular proteins. AB - Van der Waals interaction energy in globular proteins is presented by the interaction energies between regions of protein spatial structure with homogenous medium density distribution. We introduce a notion of the local medium permittivity as a function of absorptance of molecular groups with particular conformation. Proposed theory avoids shortcomings which are typical for the calculations on the basis of the pairwise additive approximation. The approach takes into account local peculiarities of protein spatial structure and physical chemical characteristics of amino acid residues and molecular groups. PMID- 10798526 TI - HELANAL: a program to characterize helix geometry in proteins. AB - A detailed analysis of structural and position dependent characteristic features of helices will give a better understanding of the secondary structure formation in globular proteins. Here we describe an algorithm that quantifies the geometry of helices in proteins on the basis of their C alpha atoms alone. The Fortran program HELANAL can extract the helices from the PDB files and then characterises the overall geometry of each helix as being linear, curved or kinked, in terms of its local structural features, viz. local helical twist and rise, virtual torsion angle, local helix origins and bending angles between successive local helix axes. Even helices with large radius of curvature are unambiguously identified as being linear or curved. The program can also be used to differentiate a kinked helix and other motifs, such as helix-loop-helix or a helix-turn-helix (with a single residue linker) with the help of local bending angles. In addition to these, the program can also be used to characterise the helix start and end as well as other types of secondary structures. PMID- 10798527 TI - Grafting of protein-protein interaction epitope. AB - Transferring the biological function of one protein to another is a key issue in understanding the structure and function relationship of proteins. We have developed a strategy for grafting protein-protein interaction epitopes. As a first step, residues at the interface of the ligand protein which strongly interact with the receptor protein were identified. Then protein scaffolds were docked onto receptor protein based on geometric complementarity. Only high docking score matches were saved. For each saved match, the scaffold protein was accepted if it had suitable positions for grafting key interaction residues of the ligand protein. These candidate residues were mutated to corresponding residues in the ligand protein at each relevant position and the mutated scaffold protein was co-minimized with receptor protein. Finally, the minimized complexes were evaluated by a scoring function deduced from statistical analysis of rigid binding data sets. As a test case, the binding epitope of barstar, the inhibitor of barnase, was grafted onto smaller proteins. Pheromone Er-1 (PDB entry 1erc) has been found to be a good scaffold. The calculated binding free energy for mutated Pheromone Er-1 is equivalent to that of barstar. PMID- 10798528 TI - A graphic approach to evaluate algorithms of secondary structure prediction. AB - Algorithms of secondary structure prediction have undergone the developments of nearly 30 years. However, the problem of how to appropriately evaluate and compare algorithms has not yet completely solved. A graphic method to evaluate algorithms of secondary structure prediction has been proposed here. Traditionally, the performance of an algorithm is evaluated by a number, i.e., accuracy of various definitions. Instead of a number, we use a graph to completely evaluate an algorithm, in which the mapping points are distributed in a three-dimensional space. Each point represents the predictive result of the secondary structure of a protein. Because the distribution of mapping points in the 3D space generally contains more information than a number or a set of numbers, it is expected that algorithms may be evaluated and compared by the proposed graphic method more objectively. Based on the point distribution, six evaluation parameters are proposed, which describe the overall performance of the algorithm evaluated. Furthermore, the graphic method is simple and intuitive. As an example of application, two advanced algorithms, i.e., the PHD and NNpredict methods, are evaluated and compared. It is shown that there is still much room for further improvement for both algorithms. It is pointed out that the accuracy for predicting either the alpha-helix or beta-strand in proteins with higher alpha-helix or beta-strand content, respectively, should be greatly improved for both algorithms. PMID- 10798529 TI - Structural equilibria in RNA as revealed by 19F NMR. AB - We have incorporated 5-fluorouridine into several sites within a 19-mer RNA modelled on the translational operator of the MS2 bacteriophage. The 19F NMR spectra demonstrate the different chemical shifts of helical and loop fluorouridines of the hairpin secondary structure. Addition of salt gives rise to a species in which the loop fluorouridine gains the chemical shift of its helical counterparts, due to the formation of the alternative bi-molecular duplex form. This is supported by UV thermal melting behaviour which becomes highly dependent on the RNA concentration. Distinct 19F NMR signals for duplex and hairpin forms allow the duplex-hairpin equilibrium constant to be determined under a range of conditions, enabling thermodynamic characterisation and its salt dependence to be determined. Mg2+ also promotes duplex formation, but more strongly than Na+, such that at 25 degrees C, 10 mM MgCl2 has a comparable duplex-promoting effect to 300 mM NaCl. A similar effect is observed with Sr2+, but not Ca2+ or Ba2+. Additional hairpin species are observed in the presence of Na+ as well as Mg2+, Ca2+, Sr2+ and Ba2+ ions. The overall, ensemble average, hairpin conformation is therefore salt-dependent. Electrostatic considerations are thus involved in the balance between different hairpin conformers as well as the duplex-hairpin equilibrium. The data presented here demonstrate that 19F NMR is a powerful tool for the study of conformational heterogeneity in RNA, which is particularly important for probing the effects of metal ions on RNA structure. The thermodynamic characterisation of duplex-hairpin equilibria will also be valuable in the development of theoretical models of nucleic acid structure. PMID- 10798530 TI - DNA duplexes containing altered sugar residues as probes of EcoRII and MvaI endonuclease interactions with sugar-phosphate backbone. AB - Oligonucleotides containing 1-(beta-D-2'-deoxy-threo-pentofuranosyl)cytosine (dCx) and/or 1-(beta-D-2'-deoxy-threo-pentofuranosyl)thymine (dTx) in place of dC and dT residues in the EcoRII and MvaI recognition site CC(A/T)GG were synthesized in order to investigate specific recognition of the DNA sugar phosphate backbone by EcoRII and MvaI restriction endonucleases. In 2' deoxyxylosyl moieties of dCx and dTx, 3'-hydroxyl groups were inverted, which perturbs the related individual phosphates. Introduction of a single 2' deoxyxylosyl moiety into a dC x dG pair resulted in a minor destabilization of double-stranded DNA structure. In the case of a dA x dT pair the effect of a 2' deoxyxylose incorporation was much more pronounced. Multiple dCx modifications and their combination with dTx did not enhance the destabilization effect. Hydrolysis of dCx-containing DNA duplexes by EcoRII endonuclease was blocked and binding affinity was strongly depended on the location of an altered sugar. A DNA duplex containing a dTx residue was cleaved by the enzyme, but kcat/K(M) was slightly reduced. In contrast, MvaI endonuclease efficiently cleaved both types of sugar-altered substrate analogs. However it did not cleave conformationally perturbed scissile bonds, when the corresponding unmodified bonds were perfectly hydrolyzed in the same DNA duplexes. Based on these data the possible contributions of individual phosphates in the recognition site to substrate recognition and catalysis by EcoRII were proposed. We observed strikingly non equivalent inputs for different phosphates with respect to their effect on EcoRII DNA complex formation. PMID- 10798531 TI - Fold-back tetraplex DNA species in DNase I-resistant DNA isolated from HeLa cells. AB - A DNase I-resistant DNA species has been isolated and purified from HeLa cells by gel electrophoresis. Our studies indicate that the DNase I-resistant DNA species was about 40-60 bp fragment sizes responding to double-strand DNA marker and has higher guanine content. The image of AFM showed that this species has been assumed to be tetraplex structure according to its apparent width and height. Its CD, UV spectrum also exhibited characteristics similar to some tetraplex structure, which was different from the standard duplex DNA. 32P-labeled probes (TTAGGG)4 and 5'-TGGGGAGGGTGGGGAGGGTGGGGAAGG-3' could be hybridized to purified DNase I-resistant species. All results suggest that the DNase I-resistant DNA species have at least two components, which adopt an intrastrand fold-back DNA tetraplex. Their sequences were similar to human telomere and human c-myc locus (NHE), respectively. PMID- 10798532 TI - Spectroscopic studies on ethidium bromide binding to intramolecular parallel and antiparallel triple helices containing T*A:T and G*G:C triplets. AB - The interaction of ethidium bromide (EB), a DNA intercalator, with two intramolecular triplexes 5'd(G4A4G4-[T4]-C4T4C4-[T4]-G4T4G4), 5'd(G4T4G4-[T4] G4A4G4-[T4]-C4T4C4) ([T4] represents a stretch of 4 thymine residues) and their precursor duplexes has been investigated by circular dichroism, fluorescence and UV absorption spectroscopy. Binding of EB induces a circular dichroism band in the region around 310 nm which is positive for the duplex forms but negative for the triplex forms. We observed that the binding of EB to the duplex form does not induce the formation of the triplex structures. Thermal denaturation experiments demonstrate that EB stabilizes more the parallel triple helix than the antiparallel one. Analysis of the binding process from fluorescence measurements shows that binding constants to the triple helical forms and to the hairpin reference duplex [T4]-G4A4G4-[T4]-C4T4C4) are close. However the binding site size is larger for the triplexes (4-6 base triplets) than for the duplex (2 base pairs). PMID- 10798534 TI - Long-range interactions between ligands bound to a DNA molecule give rise to adsorption with the character of phase transition of the first kind. AB - Influence of long-range interactions between ligands bound to DNA molecule on the character of their adsorption is studied using computer modeling. For this investigation, two calculation procedures are developed. They are based upon the method of the free energy minimum and on the partition function method. The both procedures demonstrate that in the case of a strong enough attraction between all the bound ligands their binding to DNA has the character of phase transition of the first kind. There is a break in the binding curve c(c0) where c - relative concentration of bound ligands, c0 - molar concentration of free ligands. The break occurs because there is an interval of central degrees of binding (approximately 50% of the maximum c value) that are prohibited for individual DNA molecules. Such a transition might be caused by some types of DNA condensation. Attraction between the neighboring ligands only, adjacent or/and separated by double helix regions, does not cause this effect. PMID- 10798533 TI - NMR investigation of the complexation of daunomycin with deoxytetranucleotides of different base sequence in aqueous solution. AB - 500 MHz NMR spectroscopy has been used to investigate the complexation of the anthracycline antibiotic daunomycin (DAU) with self-complementary deoxytetranucleotides, 5'-d(CGCG), 5'-d(GCGC), 5'-d(TGCA), 5'-d(ACGT) and 5' d(AGCT), of different base sequence in aqueous salt solution. 2D homonuclear 1H NMR spectroscopy (TOCSY and NOESY) and heteronuclear 1H - 31P NMR spectroscopy (HMBC) have been used for complete assignment of the non-exchangeable protons and the phosphorus resonance signals, respectively, and for a qualitative determination of the preferred binding sites of the drug. Analysis shows that DAU intercalates preferentially into the terminal sites of each of the tetranucleotides and that the aminosugar of the antibiotic is situated in the minor groove of the tetramer duplex, partly eclipsing the third base pair. A quantitative determination of the complexation of DAU with the deoxytetranucleotides has been made using the experimental concentration and temperature dependences of the drug proton chemical shifts; these have been analysed in terms of the equilibrium reaction constants, limiting proton chemical shifts and thermodynamical parameters (enthalpies deltaH, entropies deltaS) of different drug-DNA complexes (1:1, 1:2, 2:1, 2:2) in aqueous solution. It is found that DAU interacts with sites containing three adjacent base pairs but does not show any significant sequence specificity of binding with either single or double-stranded tetranucleotides, in contrast with other intercalating drugs such as proflavine, ethidium bromide and actinomycin D. The most favourable structures of the 1:2 complexes have been derived from the induced limiting proton chemical shifts of the drug in the intercalated complexes with the tetranucleotide duplex, in conjunction with 2D NOE data. It has been found that the conformational parameters of the double helix and the orientation of the DAU chromophore in the intercalated complexes depend on base sequence at the binding site of the tetramer duplexes in aqueous solution. PMID- 10798535 TI - Conformational transitions of DNA induced by changing water content of the sample: a theoretical study. AB - A semi-phenomenological model with spatially distributed parameters is suggested to describe the processes of conformational transitions induced with change of water content of wet DNA samples. It allows describing conformational dynamics of DNA molecules with heterogeneous primary structures. It has been shown that the process of cooperative conformational transition can be simulated as propagating front of a new conformation. The evolution of a conformational perturbation of DNA molecule has been described. It can collapse for finite time or occupy the whole molecule depending on the water content of the sample. PMID- 10798536 TI - Harmonic analysis of DNA dynamics in a viscous medium. AB - The harmonic dynamics of normal modes of double-stranded DNA in a viscous fluid are investigated. The model DNA consists of two backbone-supported DNA strands coiling around a common helix axis with base stacking, sugar puckering, interstrand hydrogen bonding, and intrastrand sugar-base interactions assigned values based on published data. Assuming that the DNA bases are shielded from direct bombardment by the solvent, analytical solutions are obtained. The dissipation and fluctuation of the normal modes of the bases moving along the spirals display the effect of the medium indirectly through interactions with the backbone. The dynamics of the backbone are found to be overdamped with the characteristic damping times extending to the picosecond region for disturbance in position and to the sub-picosecond region for disturbance in velocity. In addition to the dynamic mode of a rigid rod, the motions of the bases are coupled to the motions of the backbone with comparable amplitudes for disturbance in position. For disturbance in velocity, however, the bases are effectively at rest, not being able to follow the motions of the backbone. The angular frequencies of the underdamped vibrational modes, identified as the ringing modes of the bases with the backbone effectively at rest, are insensitive to the viscosity and lie in the low frequency region of the Raman spectrum. These findings indicate that the backbone of DNA plays a significant role in modulating the dynamics of double-stranded DNA in an overdamping environment. This modulation of the dynamics of the motions of the bases in DNA by environmental impediments to molecular motion is briefly discussed in connection with protein- and drug- DNA interactions as well as gene regulation. PMID- 10798537 TI - Analysis of individual specific cytotoxic T lymphocytes for two MAGE-3-derived epitopes presented by HLA-A24. AB - BACKGROUND: The human MAGE-3 gene encodes tumor-specific antigens that are recognized by cytotoxic T lymphocytes (CTLs) and expressed in a high percentage of various malignant tumors. Of the five MAGE-3-derived CTL epitopes identified to date, two nonapeptides (TFPDLESEF and IMPKAGLLI, designated MAGE-3.A24a and MAGE-3.A24b, respectively) can be expressed on the tumor surface by binding to the HLA-A24 molecule, which is the most frequent HLA class I molecule in Asian populations. To compare the immunogenecities of the two peptides, individual specific CTL lines were generated for each peptide (MAGE-3.A24a and MAGE-3.A24b). METHODS: Peripheral blood mononuclear cells (PBMCs) from four HLA-A24+ healthy donors were stimulated in vitro with autologous dendritic cells pulsed with MAGE 3.A24a, MAGE-3.A24b or both and were subsequently cultivated with a cytokine combination including interleukin-2. RESULTS: We succeeded in generating peptide specific CTL lines in two of the four donors. The two CTL lines showed similar cytolytic levels against three MAGE-3+/HLA-A24+ cancer cell lines and also target cells pulsed with the corresponding peptide. Cytolytic activities were blocked by either anti-CD8 or anti-HLA-A24 monoclonal antibodies. CONCLUSIONS: The results suggest that MAGE-3.A24a and MAGE-3.A24b peptides have equal potential in inducing MAGE-3-specific and HLA-A24-restricted CTLs. PMID- 10798539 TI - Infrequent involvement of the anterior base in low-risk patients with clinically localized prostate cancer and its possible significance in definitive radiation therapy. AB - BACKGROUND: The zonal distribution and location of tumors in different subgroups of Japanese patients with clinically localized prostate cancer have not been fully described. The appropriate radiation treatment volume thus remains unclear. METHODS: Radical prostatectomy specimens of 141 consecutive patients with clinically localized prostate cancer were examined by the whole organ step section technique. The zonal distribution and location of tumors at different levels of the gland were investigated after stratification into patient subgroups based on preoperative clinicopathological findings and risk group assessment. RESULTS: The median tumor volume was 2.8 cm3; 72 patients (51.1%) had pathologically organ-confined disease (pT2). Higher risk groups showed a statistically significant increase in tumor volume and a decrease in the rate of pathologically confirmed organ confinement. Involvement of the anterior base was found infrequently in certain patient subgroups: in only one of 20 patients (5%) with preoperative PSA <4.0 ng/ml, in three of 19 patients (15.8%) with specimen Gleason scores of 2-4 and in two of 32 patients (6.3%) identified as low-risk. CONCLUSIONS: Infrequent involvement of the anterior base in low-risk patients may be an intrinsic feature of clinically localized prostate cancer. Treatment volume modifications in these patients that reduce the radiation dose to the anterior base may be justified to avoid acute and late genitourinary toxicities. PMID- 10798538 TI - Successful initial treatment with weekly etoposide, epirubicin, cisplatin, 5 fluorouracil and leucovorin chemotherapy in advanced gastric cancer patients with disseminated intravascular coagulation. AB - BACKGROUND: Acute disseminated intravascular coagulation (DIC) is a rare but severe complication of gastric adenocarcinoma. Conventional treatments, such as fresh frozen plasma, platelet replacement and heparin injections, are disappointing. The only way to correct this fatal condition is to control the underlying cancer promptly by effective chemotherapy. Here the successful initial control of acute DIC in gastric cancer patients with weekly EEPFL chemotherapy is reported. METHODS: Advanced gastric cancer patients complicated with acute DIC were eligible. Patients were treated with weekly EEPFL therapy (etoposide 40, epirubicin 10, cisplatin 25, 5-fluorouracil 2200 and leucovorin 120 mg/m2 ). Response, survival and toxicity were evaluated. RESULTS: From April 1997 to April 1999, six patients were included in this study. All patients received EEPFL chemotherapy. Clinical and laboratory evidence of acute DIC stabilized quickly after starting chemotherapy. Four patients showed a partial response, one stable disease and one progressive disease. The toxicity was mild and well tolerated. Median survival was 28 weeks (12, 14, 26, 30, 30 and 32 weeks). All patients suffered from a relapse of DIC after initial successful control and died within 30 days of clinical and laboratory evidence of acute DIC relapse. CONCLUSION: EEPFL therapy is an effective chemotherapy regimen for patients with advanced gastric cancer associated with acute DIC. The prognosis is poor if the DIC relapses after the initial successful control. PMID- 10798540 TI - Early results of LH-RH agonist treatment with or without chlormadinone acetate for hormone therapy of naive localized or locally advanced prostate cancer: a prospective and randomized study. The Prostate Cancer Study Group. AB - BACKGROUND: The majority of patients with localized and some cases of locally advanced prostate cancer undergo radical prostatectomy. However, radical prostatectomy cannot always be selected for those patients. In this situation, primary hormone therapy is an alternative treatment option. We have designed a prospective randomized study of the effects of primary hormone therapy for such patients. METHODS: A total of 151 patients with T1b, T1c, T2a, T2b or T3a prostate cancer who were not scheduled for radical prostatectomy were enrolled into this study. Patients were randomly allocated into two groups; Group I received luteinizing hormone-releasing hormone (LH-RH) agonist monotherapy (leuprorelin acetate depot, 3.75 mg monthly) and Group II received LH-RH agonist in combination with chlormadinone acetate (100 mg/day). Effects on serum prostate specific antigen level, progression-free survival and survival were observed for 2 years. RESULTS: The reasons why radical prostatectomy was not scheduled were poor risk for surgery (38%), patient's wish (32%) and physician's recommendation (30%). After 12 weeks of treatment, 49% of the patients in both groups showed a complete response (CR). Of the patients showing a partial response (PR) after 12 weeks of treatment, 25% in Group I and 52% in Group II improved to CR 1 year later (p<0.05). Group II showed a longer progression-free survival (p <0.05). Progression-free survival rates were 62% (Group I) and 91% (Group II) in T2b patients and 43% (Group I) and 73% (Group II) in T3 patients. Only one patient in each group died from prostate cancer. CONCLUSIONS: Early primary hormone therapy is a reasonable treatment option for localized or locally advanced prostate cancer patients if radical prostatectomy was not scheduled. Chlormadinone acetate showed an additive effect with LH-RH agonist, at least in 2 years' observation. PMID- 10798541 TI - Feasibility study of autologous peripheral blood stem cell transplantation for the treatment of childhood acute myelogenous leukemia. AB - BACKGROUND: The primary object of this study was to identify treatment-related variables that may predict relapse of acute myelogenous leukemia (AML) after autologous peripheral blood stem cell transplantation (PBSCT), which will be critical for the development of a suitable protocol for wider application. METHODS: A total of 28 children (age 0-18 years) with AML underwent PBSCT and have had a minimum follow-up of 25 months; including 24 patients in their first complete remission (CR) and four in their second CR. The patients were divided into two cohorts according to the study phase: 16 patients were treated in an early phase pilot study and 12 patients in their first CR were treated in a prospective trial. Fifteen of the first-CR patients had any of the cited high risk features of high WBC count (>100x10(9)/l; n = 5), FAB M0/M4/M5/M7 subtypes (n = 11) or delayed achievement of CR (n = 9). Except in one patient, cytoreductive regimens did not include total body irradiation (TBI). RESULTS: After PBSCT, one patient died of veno-occulsive disease (VOD) and another patient relapsed early on day 43, but the remaining patients showed engraftment. Leukemic relapse was observed 1-29 months after PBSCT (median, 8 months); in all of the 4 children treated in their second CR and in 11 of the 24 patients (46%) treated in their first CR. The remaining patients have been disease-free for 24 to 97 months (median, 53 months). Using a multivariate analysis, the timing of apheresis was the most significant prognostic factor for those treated in their first CR (p = 0.03); 12 of the 16 patients whose PBSC were collected beyond 2.5 months of CR continue to remain in CR, while seven of the eight patients whose PBSC were harvested within 2.5 months of CR relapsed. CONCLUSION: Although the small number of patients studied does not allow firm conclusions to be drawn regarding the relative effectiveness of this therapy, the results do suggest the feasibility of further studies of PBSCT for the treatment of childhood AML with high-risk features including the assessment of minimum residual disease. PMID- 10798542 TI - Alternating combination chemotherapy C-MOPP (cyclophosphamide, vincristine, procarbazine, prednisone) and ABVd (adriamycin, bleomycin, vinblastine, dacarbazine) in clinical stage II-IV Hodgkin's disease: a multicenter phase II study (JCOG 8905). The Lymphoma Study Group of the Japan Clinical Oncology Group. AB - BACKGROUND: The main form of cytotoxic treatment for advanced Hodgkin's disease (HD) is conventional dose multiagents chemotherapy. As HD is not common in Japan, we conducted a phase II study of the commonly used combination chemotherapy (CCT) regimen established in the West for Japanese patients with advanced HD to confirm the efficacy and safety. METHOD: Between October 1989 and February 1993, a multicenter phase II study of alternating CCT C-MOPP (cyclophosphamide, vincristine, procarbazine, prednisone) and ABVd (adriamycin, vinblastine, bleomycin, dacarbazine) to evaluate its clinical usefulness for clinical stage (cS) II-IV HD was conducted by the Lymphoma Study Group of the Japan Clinical Oncology Group. RESULTS: Seventy-nine previously untreated patients were enrolled in the study. For 67 eligible patients, the response rate was 92.5% with 83.6% complete response (CR). For 40 cS II and 27 cS III/IV patients the response rate was 95.0% with 90.0% CR and 88.9% with 74.1% CR, respectively. The overall 5-year survival rate was 84.8%. Those of cS II and cS III/IV were 92.5 and 73.1%, respectively. There was no significant difference between cS II and cS III/IV (p = 0.1025). The progression-free 4-year survival rate was 72.8%. Those of cS II and cS III/IV were 77.5 and 65.7%, respectively. There was no significant difference between cS II and cS III/IV (p = 0.2483). Grade 4 toxicity by the criteria of the World Health Organization consisted of leukocytopenia in 28.4% of patients. There was GPT elevation in 4.5%, nausea/vomiting in 11.9% and CNS in 1.5% of patients, but there was no treatment-related death. CONCLUSION: The C MOPP/ABVd regimen for Japanese patients with advanced HD is considered to be one of the effective CCTs according to the results of the present phase II study. PMID- 10798543 TI - A successful and simplified filgrastim primed single apheresis method without large volume apheresis for peripheral blood stem cell collection. AB - BACKGROUND: There is a tendency to use only one apheresis collection to reduce the morbidity and the cost of peripheral blood stem cell collection. We studied whether rapid and complete engraftment could be achieved by single apheresis by using only Filgrastim without large volume apheresis in previously treated patients. METHODS: Engraftment of single apheresis in 25 patients was compared with those of multiple apheresis in 26 patients; 52% of patients in the single apheresis group and 62% of patients in the multiple apheresis group were heavily pretreated. All patients received 10-15 microg/kg/day of Filgrastim starting on day 14 after 3-4 cycles of induction chemotherapy. Apheresis was performed using Cobe Spectra on day 4, 5 or 6 in the single apheresis group and every other day in the multiple apheresis group after day 3. RESULTS: The median collection volume was 250 ml (250-300 ml) in the single apheresis group and 750 ml (200-1500 ml) in the multiple apheresis group. The median CD34(+) cell number was not significantly different in the two groups (11.79 vs. 9.38x10(6)/kg). The median times to achieve leukocytes > or =1x10(9)/l and platelets > or =50x10(9)/l counts were 10 days (8-21 days) and 15 days (9-38 days) in the single apheresis group vs 11 days (8-23 days) and 20 days (10-32 days) in the multiple apheresis group, respectively (p<0.05). Antibiotic use was less in the single apheresis group than the multiple apheresis group (9 vs. 12 days, p<0.05). CONCLUSION: Adequate numbers of peripheral stem cells were harvested by G-CSF in a single apheresis without large volume apheresis even in heavily pretreated patients. Rapid and complete engraftment occurred in all patients and it was faster in single than multiple apheresis. PMID- 10798544 TI - A case of bilateral heel ulcers associated with hydroxyurea therapy for chronic myelogenous leukemia. AB - Bilateral heel skin ulcers developed in a 50-year-old male in the chronic phase of chronic myelogenous leukemia who had been receiving hydroxyurea (HU) therapy for 3 years. Histological examination showed perivascular lymphocytic inflammation without vasculitis. After interruption of HU administration, the heel ulcers were completely resolved within 2 months. The clinical course strongly suggested that the heel ulcers were induced by long-term HU therapy. PMID- 10798546 TI - Diabetes and cancer: is there any relationship between two diseases? PMID- 10798545 TI - Secretion of hCG/beta-hCG by squamous cell carcinoma of the lung in a 31-year-old female smoker. AB - We describe a rare case of pulmonary squamous cell carcinoma secreting human chorionic gonadotropin (hCG) and its beta-subunit (beta-hCG) in a young female smoker. A 31-year-old mother of one child had been suffering from dysfunctional uterine bleeding for about 1 year. Pelvic examinations and abdominal ultrasonography yielded no abnormal findings and no signs of pregnancy. She developed a pain in the right chest and a huge (12x10 cm) squamous cell carcinoma was diagnosed in the right lower lobe. The serum hCG and beta-hCG levels were high: hCG 5611 mlU/ml (normal upper limit 0.7 mIU/ml), beta-hCG 12 238 mIU/ml (normal upper limit 0.5 mIU/ml). The patient underwent right lower lobectomy and systematic lymph node dissection. Microscopic study showed a poorly differentiated squamous cell carcinoma. The pathological stage was T2N0M0, stage IB. Immunohistochemical staining of the tumor was strongly positive for hCG. The serum hCG level returned to normal 1 month after the operation, while an additional 2 months were necessary for the beta-hCG level to normalize. Dysfunctional uterine bleeding disappeared and the patient is doing well, with no signs of recurrence, 9 months after the resection. PMID- 10798547 TI - Cancer Statistics Digest. All cancer mortality by prefectures in Japan PMID- 10798548 TI - European Organization for Research and Treatment of Cancer. PMID- 10798549 TI - Rhinoscleroma. PMID- 10798550 TI - Strain, clone and species: comments on three basic concepts of bacteriology. AB - Different aspects of the terms strain, clone and species are discussed. The term strain is commonly used to denote a pure culture - here called 'the strain in the taxonomic sense' - but does also refer to a natural concept closely related to the clone. The term clone on the other hand is used both in a general and in a more restricted sense, the latter indicating a low degree of genetic exchange. The important distinction between the definition of a species and the criteria for a species is emphasised and the main kinds of criteria are considered. PMID- 10798551 TI - Beta-lactamase expression in Yersinia enterocolitica biovars 1A, 1B, and 3. AB - Characteristic patterns of beta-lactam susceptibility are associated with different biovars of Yersinia enterocolitica. In a previous study differences in beta-lactam susceptibility among biovar 2, 4 and 5 strains were largely attributed to differences in expression of beta-lactamase A (BlaA) and beta lactamase B (BlaB). The basis for differences in beta-lactam susceptibility of strains of biovars 1A, 1B and 3 is now considered. All the strains examined had blaB; nine of 31 biovar 3 strains and two of 13 biovar 1B strains had blaA, but PCR did not amplify blaA from biovar 1A strains. Nevertheless, inhibition data indicated that the majority of uninduced biovar 1A strains expressed BlaA and BlaB in similar amounts. Strong inducibility was seen in all these strains. Biovar 1B strains (which were less inducible than strains of biovar 1A) predominantly produced BlaA without induction; ticarcillin-sensitive strains of biovar 3 produced only BlaB but were not inducible; without induction biovar 3 strains resistant to ticarcillin and amoxycillin/clavulanate produced either predominantly BlaA, predominantly BlaB or exclusively BlaB and induction was demonstrated except for strains producing BlaB alone; biovar 3 strains resistant to ticarcillin but sensitive to amoxycillin/clavulanate predominantly produced BlaA without induction and were inducible for beta-lactamase activity. After induction, nearly all strains predominantly or exclusively produced BlaB. Although PCR amplification fragments with primers specific for blaA were obtained only from some strains, the induction and inhibition data suggest that all Y. enterocolitica strains possess enzymes related to BlaA- as well as BlaB. Nevertheless, expression of the beta-lactamase is regulated differently in different biovars and varies within most biovars. Failure to predict beta lactamase expression profiles from MIC data indicates the presence of additional mechanisms contributing to differences in susceptibility. PMID- 10798552 TI - Altered expression of oligopeptide-binding protein (OppA) and aminoglycoside resistance in laboratory and clinical Escherichia coli strains. AB - Oligopeptide-binding protein (OppA) is the periplasmic component of the major oligopeptide transport system of enteric bacteria. Genetic and biochemical evidence suggests that OppA plays a role in the uptake of aminoglycoside antibiotics in Escherichia coli K-12. Forty-six (82%) of 56 aminoglycoside resistant mutants of E. coli K-12 selected in vitro had reduced or undetectable OppA levels, as compared with their parent strain. Moreover, nine (36%) of 25 aminoglycoside-resistant clinical isolates of E. coli expressed reduced or undetectable levels of OppA. No decrease in OppA expression was observed among aminoglycoside-sensitive E. coli strains from patients. Twenty-three (42%) of 56 aminoglycoside-resistant mutants of E. coli K-12 and six (24%) of 25 clinical isolates also were deficient for expression of ornithine or arginine decarboxylases, or both, and these deficiencies might negatively affect OppA expression by reducing polyamine synthesis. These results support the view that reduced OppA expression is associated with aminoglycoside resistance in E. coli strains. PMID- 10798553 TI - Primary and combined resistance to four antimicrobial agents in Helicobacter pylori in Sofia, Bulgaria. AB - The aim of this study was to evaluate the primary and combined resistance of Helicobacter pylori against four antimicrobial agents by a screening agar method (SAM) and a modified disk diffusion method (MDDM) alone and in combination. Pre treatment H. pylori isolates from 192 consecutive H. pylori-positive patients at three hospitals in Sofia were investigated. MDDM was performed with disks containing metronidazole (5 microg), clarithromycin (15 microg) or erythromycin (15 microg), ciprofloxacin (5 microg) and tetracycline (30 microg). Resistance was determined by an inhibitory zone of <16 mm for metronidazole and < or =30 mm for other agents tested. The cut-off concentrations used to define resistance by SAM were: metronidazole >8 mg/L, clarithromycin >2 mg/L, tetracycline >4 mg/L and ciprofloxacin >1 mg/L. Primary resistance rates in H. pylori were: metronidazole 28.6%, clarithromycin 9.7%, metronidazole + clarithromycin 2.8%, ciprofloxacin 3.9%, metronidazole + ciprofloxacin 2.3%, tetracycline 1.9% and metronidazole + tetracycline 1.2%. Among metronidazole-resistant isolates, combined resistance to clarithromycin, ciprofloxacin and tetracycline was present in 11.4% (5 of 44 strains), 8.3% (3 of 36) and 4.9% (2 of 41), respectively. Two strains exhibited triple resistance to macrolides, metronidazole and either ciprofloxacin or tetracycline. Three tetracycline-resistant strains were detected in 1999; however, resistance rates to other agents were relatively stable during the 6 years. Primary H. pylori resistance to metronidazole is moderate and resistance to clarithromycin and to ciprofloxacin is considerable in comparison with results in most other countries. The alarming appearance of strains harbouring combined resistance or multiresistance provides the motivation for continued surveillance of H. pylori at global, national and regional levels. PMID- 10798554 TI - Characterisation of methicillin-resistant Staphylococcus aureus from Kuwait hospitals with high-level fusidic acid resistance. AB - Forty-seven fusidic acid- and methicillin-resistant Staphylococcus aureus isolates from clinical samples in four hospitals in Kuwait were studied for their relatedness by biotyping and pulsed-field gel electrophoresis (PFGE) and for the genetic location of their resistance determinants. Forty-four isolates were resistant to gentamicin, kanamycin and neomycin. Forty-one isolates were resistant to erythromycin and trimethoprim, 10 were resistant to chloramphenicol and four were resistant to ciprofloxacin. They contained two or three plasmids of c. 28, 2.8 and 1.8 kb. Genetic studies demonstrated that resistance to cadmium, propamidine isethionate and ethidium bromide were linked and were carried on the c. 28-kb plasmid. Chloramphenicol resistance was encoded by the 2.8-kb plasmid in resistant isolates. No resistance was associated with the 1.8-kb plasmid and this was considered to be a cryptic plasmid. Resistance to fusidic acid, methicillin, benzylpenicillin, gentamicin, kanamycin, neomycin, tetracycline, trimethoprim, erythromycin and ciprofloxacin were located on the chromosome. All the isolates produced urease, but varied in the production of haemolysins, pigments, lipase and lecithinase and were classified into nine biotypes. In contrast, PFGE divided the isolates into two major patterns with one PFGE type constituting the majority of isolates in all four hospitals. The presence of the dominant PFGE pattern in all four hospitals suggests that it is an epidemic MRSA clone with the capacity to spread. Infection control measures should be directed towards restricting the further spread of this clone. PMID- 10798555 TI - Prevalence of the carbapenemase gene (cfiA) among clinical and normal flora isolates of Bacteroides species in Hungary. AB - The carbapenemase gene (cfiA) was detected in 4 (5.7%) of 70 clinical isolates of Bacteroides fragilis from different parts of Hungary. Among 24 other Bacteroides species isolated from infectious processes or from normal faecal flora, none was cfiA-positive. The MIC of imipenem and meropenem for all cfiA-positive B. fragilis isolates was < or =0.25 mg/L, but 17% of the B. fragilis and 46% of the non-fragilis Bacteroides isolates exhibited reduced susceptibility to imipenem (MICs 0.5-2 mg/L). Only one of these isolates produced increased levels of beta lactamase. No difference was observed in the outer-membrane proteins of B. fragilis isolates that harboured the cfiA gene and those with reduced susceptibility to imipenem. PMID- 10798556 TI - Translocation of indigenous microflora in an experimental model of sepsis. AB - Translocation of viable bacteria from gut to bloodstream and other sterile body sites during shock has been demonstrated in several experimental and clinical studies. The factors causing translocation and its incidence at different stages of shock are not known. The aim of the study was to evaluate the importance of several factors causing translocation of indigenous microflora in an experimental model of septic shock based on intraperitoneal Escherichia coli sepsis in rats. Counts of inoculated E. coli and translocated bacteria in different locations, gut morphology and haematological values were evaluated at different stages of sepsis. Sepsis developed in all animals and E. coli achieved the highest counts in blood 6 h after inoculation. Translocation was commonest at 6 and 12 h after inoculation. Frequently translocating bacteria were lactobacilli, bifidobacteria, bacteroides and peptostreptococci. In early sepsis, translocation was associated with high E. coli counts in blood, yet in late sepsis the opposite correlation was present. Low infiltration by neutrophils in the ileum and decreased mitotic activity in the colon were associated with a high translocation rate. In early sepsis, translocation was associated with low lymphocyte counts, but in late sepsis, with low neutrophil counts. Translocation of bacteria (including anaerobes) that colonise the gut in high counts takes place during sepsis. Putative influencing factors such as activity of the primary disease (bacterial counts in blood), gut morphology or haematological values seem to have different impacts on translocation, depending on the stage of the disease. PMID- 10798557 TI - Use of scanning electron microscopy to investigate the prophylactic efficacy of rifampin-impregnated CSF shunt catheters. AB - Infection continues to be one of the major complications of cerebrospinal fluid (CSF) shunting procedures, and is caused mainly by skin-derived bacteria. Production of an extracellular biofilm plays an important role in the pathogenesis of shunt-associated infections by protecting bacteria from immune mechanisms and antibiotics. So far, removal of the original shunt and implantation of a new shunting device has been the only successful treatment for most patients. As an alternative strategy to prevent CSF infections, a rifampin impregnated silicone catheter was designed to provide high initial and long lasting (>60 days) release of bactericidal drug. To investigate the pathophysiological mechanism of its function, this new device was investigated both in vitro and in a rodent model of CSF infection by scanning electron microscopy (SEM) and bacterial culture. Staphylococcus epidermidis (10(8) cfu/ml) and S. aureus (10(4) cfu/ml) served as test strains. SEM demonstrated that, in contrast to the unloaded catheters, initial bacterial adherence on the catheter surface could be reduced to a few single cells, which did not show visible signs of proliferation. Bacterial cultures obtained simultaneously were all sterile, showing that adherent bacteria were killed immediately by the rifampin released from the catheter. Although rifampin incorporation into silicone polymers was not able to prevent initial bacterial adhesion completely, subsequent colonisation could be prevented. PMID- 10798558 TI - Evaluation of a PCR-immunoassay technique for detection of Neisseria meningitidis in cerebrospinal fluid and peripheral blood. AB - A multiplex PCR-immunoassay for the rapid diagnosis of bacterial meningitis from cerebrospinal fluid (CSF) or peripheral blood was compared with conventional microbiological techniques used in the routine diagnostic laboratory. The multiplex PCR was designed to detect simultaneously a universal conserved sequence coding for bacterial 16S rRNA and the Neisseria meningitidis porA gene. The PCR-immunoassay had a detection limit of (3-5) x 10(2) cfu/ml (equivalent to 1-3 cfu/PCR) with spiked CSF or blood samples, compared with (3-5) x 10(3) cfu/ml for PCR followed by conventional agarose gel electrophoresis for detection of PCR products. Of 294 CSF samples from patients suspected on clinical grounds of suffering from meningitis, the PCR-immunoassay detected bacterial DNA in 29 CSF samples, 15 of which were also positive for N. meningitidis DNA. The 29 positive CSF samples comprised 25 samples that were also reported positive and four that were reported negative by conventional methodology; the latter four were all positive for N. meningitidis by the PCR-immunoassay. Of 173 peripheral blood samples examined, the PCR-immunoassay detected bacterial DNA in 18 samples, 14 of which were also positive for N. meningitidis DNA. In comparison, only 10 samples were reported positive for N. meningitidis by conventional methodology. All negative PCR-immunoassay results correlated with those obtained by conventional methodology for both CSF and blood samples. The sensitivity and speed of the PCR immunoassay system indicated that it could be used as a routine diagnostic test for meningococcal meningitis, enabling a diagnosis to be made within 4 h of receipt of the specimen. PMID- 10798559 TI - Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens. AB - This study was performed to generate and evaluate recombinant antigens for use in a species-specific Chlamydia trachomatis immunoassay. In a molecular genetic approach, fragments of the C. trachomatis major outer-membrane protein (MOMP) were produced as fusion proteins to create three different constructs encompassing the variable domains I, II and IV of selected C. trachomatis serovars. The recombinant MOMP polyantigens were affinity-purified and used in an enzyme-linked immunosorbent assay. Antibody detection was evaluated with 103 patient sera and the results were compared with titres obtained in the micro immunofluorescence test. The results showed that the generated MOMP polyantigens detected the presence of C. trachomatis-specific human antibodies with little cross-reaction to C. pneumoniae-specific antibodies. When compared to the micro immunofluorescence assay the MOMP polyantigen detected the presence of anti-C. trachomatis IgG antibodies with a sensitivity of 80% and a specificity of 91%. PMID- 10798560 TI - Molecular characterisation of group A streptococci from invasive and non-invasive disease episodes in Belgium during 1993-1994. AB - Five hundred clinical group A streptococcal (GAS) isolates were collected in Belgium during the period 1 Nov. 1993 to 31 Oct. 1994. Clinical and laboratory data were recorded and isolates were characterised. The presence of the genes encoding streptococcal pyrogenic exotoxin types A (speA), B (speB), C (speC), F (speF) and streptococcal superantigen (ssa) were determined by PCR to target specific sequences. These isolates were also emm-typed and analysed by pulsed field gel electrophoresis (PFGE) of genomic macrorestriction fragments with the enzyme SmaI. In total, 136 unrelated GAS PFGE types were identified and genetic diversity was clearly demonstrated. Two GAS PFGE types predominated; a first PFGE type comprised 66 (13.2%) emm1 isolates characterised by speA-, speB+, speC-, speF+ and ssa-; the second PFGE type comprised 44 (8.8%) emm12 isolates characterised by speA-, speB+, speC+ (or speC-), speF+ and ssa-. Indistinguishable PFGE types were observed among both invasive and non-invasive isolates. Ten different PFGE types were found among 11 streptococcal toxic shock syndrome (STSS) isolates, and five of these lacked speA. Twenty-five (34.7%) of 72 invasive isolates gave negative results for speA, speC and ssa. This retrospective study confirmed the observation that the dissemination of one specific clone cannot be associated with invasive GAS disease and posed a question regarding the role of SPE A as a major virulence factor. Other streptococcal virulence factors in conjunction with host factors may determine the outcome of invasive GAS infection. PMID- 10798561 TI - Characterisation of cytolethal distending toxin (CDT) mutants of Campylobacter jejuni. AB - In order to assess the contribution of cytolethal distending toxin (CDT) to the toxigenicity and pathogenicity of Campylobacter jejuni, the C. jejuni 81-176 and C. jejuni NCTC 11168 CDTs were inactivated by insertional mutation of the cdtB toxin subunit. Cell-free sonicates from isogenic C. jejuni 81-176 cdtB- strains were found to be greatly attenuated in HeLa cytotoxicity assays, whilst still retaining some toxigenicity. Sonicates from a C. jejuni NCTC 11168 cdtB- strain produced no detectable cytotoxicity. When orally administered to adult severe combined immunodeficient (SCID) mice, C. jejuni cdtB mutant strains were unaffected in enteric colonisation abilities but demonstrated impaired invasiveness into blood, spleen and liver tissues. These data suggest that CDT may be the principal toxin produced by this species and that some C. jejuni strains may generate additional toxigenic factor(s) distinct from CDT. PMID- 10798562 TI - The role of SEF14 and SEF17 fimbriae in the adherence of Salmonella enterica serotype Enteritidis to inanimate surfaces. AB - To gain an understanding of the role of fimbriae and flagella in the adherence of Salmonella enterica serotype Enteritidis to inanimate surfaces, the extent of adherence of viable wild-type strains to a polystyrene microtitration plate was determined by a crystal violet staining assay. Elaboration of surface antigens by adherent bacteria was assayed by fimbriae- and flagella-specific ELISAs. Wild type Enteritidis strains adhered well at 37 degrees C and 25 degrees C when grown in microtitration wells in Colonisation Factor Antigen broth, but not in other media tested. At 37 degrees C, adherent bacteria elaborated copious quantities of SEF14 fimbrial antigen, whereas at 25 degrees C adherent bacteria elaborated copious quantities of SEF17 fimbrial antigen. Non-fimbriate and non-flagellate knock-out mutant strains were also assessed in the adherence assay. Mutant strains unable to elaborate SEF14 and SEF17 fimbriae adhered poorly at 37 degrees C and 25 degrees C, respectively, but adherence was not abolished. Non-motile mutant strains showed reduced adherence whilst type-1, PEF and LPF fimbriae appeared not to contribute to adherence in this assay. These data indicate that SEF17 and SEF14 fimbriae mediate bacterial cell aggregation on inanimate surfaces under appropriate growth conditions. PMID- 10798563 TI - Determinants and analysis of blood alcohol concentrations after social drinking. PMID- 10798564 TI - Aspects of in-vivo pharmacokinetics of ethanol. PMID- 10798565 TI - Effect of food on the pharmacokinetics of alcohol absorption. PMID- 10798566 TI - Site and quantitative importance of alcohol first-pass metabolism. PMID- 10798567 TI - Metabolism of alcohol by the human stomach. PMID- 10798568 TI - Use of a two-compartment model to predict ethanol metabolism. PMID- 10798569 TI - Interaction of prandial state and beverage concentration on alcohol absorption. PMID- 10798570 TI - Effects of food and body composition on blood alcohol curves. PMID- 10798571 TI - Variation in ethanol pharmacokinetics and perceived gender and ethnic differences in alcohol elimination. PMID- 10798572 TI - Ethnic and gender differences in ethanol metabolism. PMID- 10798573 TI - Alcohol elimination: faster in women? PMID- 10798574 TI - Influence of sex hormones on alcohol metabolism. PMID- 10798575 TI - Transdermal alcohol measurement for estimation of blood alcohol concentration. PMID- 10798576 TI - Pharmacokinetics of intravenous alcohol: two compartment, dual Michaelis-Menten elimination. PMID- 10798577 TI - PBPK modeling as a basis for achieving a steady BrAC of 60 +/- 5 mg% within ten minutes. PMID- 10798578 TI - The infant rat learns about alcohol through interaction with an intoxicated mother. AB - BACKGROUND: Infant rats detect the presence of alcohol in milk when the dam suffers a moderate state of alcohol intoxication. The present study examined when rat pups begin to show behavioral changes indicative of the interaction with an intoxicated dam. The study also attempted to determine if infantile experiences involving a moderately intoxicated dam result in alcohol-derived memories with a particular hedonic content. METHODS: Infant rats were allowed to interact during postnatal days (PDs) 3, 5, 7, 9, 11 and 13 with alcohol-intoxicated (EtOH dose: 2.5 g/kg) or alcohol-free dams. After the interaction took place, some pups were tested in terms of ultrasonic vocalizations (USVs) and motor reactivity when isolated and placed on each of two distinctive tactile surfaces (sandpaper or soft fabric) presented in a counterbalanced order, and the second of which was always paired with ambient ethanol odor. At PD 14 pups were evaluated in terms of the preference for texture (sandpaper versus soft fabric) and odor (alcohol versus clove) as well as alcohol ingestion. RESULTS: Very early in life (PD 3) USVs and overall activity were significantly higher in pups that had previously interacted with an intoxicated dam than in those exposed to an alcohol-free dam. Although this difference was not apparent during the following days, it was clear that a specific memory of alcohol's chemosensory cues was formed. Pups interacting with intoxicated dams followed by pairing of ethanol odor and an arousing texture (sandpaper), later avoided this texture in the preference test; pups that interacted with alcohol-free dams did not show this effect. The former animals also exhibited less consumption of alcohol than preweanlings never exposed to alcohol in the context of nursing. CONCLUSIONS: In conjunction with prior studies these results indicate that very early in ontogeny the infant processes the presence of ethanol, and perhaps its effect on its mother, within the nursing context. Under the present experimental circumstances infants appear to acquire alcohol-related information that comprises an aversive hedonic component. PMID- 10798579 TI - Impairment in eyeblink classical conditioning in adult rats exposed to ethanol as neonates. AB - BACKGROUND: Eyeblink classical conditioning is a learning task that engages well defined neural circuitry in the cerebellum and brainstem. Binge-like exposure to alcohol during the neonatal brain growth spurt in rats produces neurotoxic effects on both the cerebellum and the brainstem. The precise localization of the neural substrates of eyeblink conditioning makes it an ideal task to study functional disruptions in the cerebellum and brainstem caused by early exposure to alcohol. The purpose of this study was to determine whether impairments in eyeblink conditioning caused by neonatal binge exposure to alcohol persist into adulthood, indicative of long-lasting abnormalities in cerebellar and brainstem function. METHODS: Group Ethanol received alcohol doses of 5.25 g/kg/day via intragastric intubation on postnatal days 4-9. Group Sham Intubated underwent the intragastric intubation procedures on postnatal days 4-9 but did not receive any infusions. Group Nonintubated did not receive any intubations. When all rats were at least 3 months old, they were tested in either paired or unpaired eyeblink conditioning. RESULTS: Group Ethanol showed impaired eyeblink conditioning and some abnormalities in conditioned response timing. Control groups did not differ from each other. CONCLUSIONS: The present data indicate that early exposure to alcohol has long-term effects on eyeblink conditioning, perhaps through enduring effects associated with alcohol-induced loss of Purkinje cells of the cerebellum. PMID- 10798580 TI - Individual differences predictive of drinking to manage anxiety among non-problem drinkers with panic disorder. AB - BACKGROUND: Alcohol problems co-occur with anxiety disorders at a rate that far exceeds chance ("comorbidity"). One view suggests that risk for developing a comorbid alcohol use disorder is increased when alcohol is used routinely to cope with anxiety symptoms ("self-medication"). Indicating that this view is overly broad, however, the literature suggests that only a subgroup of anxiety disordered individuals tend to drink to manage their symptoms. Therefore, we set out to identify psychological characteristics that might mark those for whom drinking to cope with anxiety is most likely. Our survey of the literature identified several possibilities, including anxiety-related personality traits (anxiety sensitivity, self-consciousness and Trait Anxiety); higher-order personality dimensions (Harm Avoidance, Reward Dependence, and Novelty Seeking); and, finally, alcohol outcome expectancies (specifically, those related to tension-reducing effects from alcohol). METHODS: In a sample of nonproblem drinkers with panic disorder, we regressed predictor variables on several alcohol use measures, including drinking aimed at the control of anxiety symptoms ("self medication"). RESULTS: Although each variable related positively to a self medicating style of drinking, expectancies for tension reduction from alcohol use accounted for about twice as much variance as did the other predictors. With simultaneous entry in a step-wise regression analysis, only tension-reduction alcohol outcome expectancies and the Harm Avoidance personality dimension were retained as significant predictors. CONCLUSIONS: Findings suggest that development of a self-medicating style of drinking among individuals with panic disorder is predicated, in part, on specific psychological characteristics of the individual. Alcohol outcome expectancies emerged as the single most important predictor of drinking behavior in this anxiety-disordered sample. PMID- 10798581 TI - Acute tyrosine depletion and alcohol ingestion in healthy women. AB - BACKGROUND: Recently we reported that, in vervet monkeys, ingestion of an amino acid mixture deficient in the catecholamine precursors, phenylalanine and tyrosine, produced a decrease in alcohol self-administration. We now report the results of a similar study in humans. METHODS: Three groups of healthy female social drinkers were administered a nutritionally balanced amino acid mixture (B, n = 13), a mixture deficient in the serotonin precursor, tryptophan (Trp-free, n = 14), or a phenylalanine/tyrosine deficient mixture (Phe/Tyr-free, n = 12). Six hours after administration of the amino acid mixture, alcohol ingestion was measured during a free-choice "Taste Test." RESULTS: Compared to the B mixture, Phe/Tyr-free, but not Trp-free, significantly decreased the ingestion of alcohol [p < 0.02]. Neither Phe/Tyr-free nor Trp-free significantly decreased orange juice ingestion or the self-reported "Liking" of either substance. Some subjects experienced transient nausea and/or regurgitated the amino acid mixtures, but excluding these subjects did not change the results. CONCLUSIONS: The results suggest that (a) Phe/Tyr-free may be a suitable method for investigating the role of catecholamines in the self-administration and subjective effects of alcohol, (b) acutely decreased catecholamine neurotransmission might disrupt mechanisms mediating alcohol self-administration, and (c) acutely decreased serotonin neurotransmission seems not to alter alcohol self-administration. PMID- 10798582 TI - Alcohol-induced brain pathology and behavioral dysfunction: using an animal model to examine sex differences. AB - BACKGROUND: Human epidemiological studies suggest that the female brain may be more susceptible to the toxic effects of alcohol and that this is the reason why women show greater behavioral dysfunction after chronic alcohol exposure. This hypothesis was tested by using a rat model of chronic alcoholism [chronic ethanol treatment (CET)]. The investigation assessed sex differences in neuropathology and behavior after chronic exposure and subsequent withdrawal from alcohol. METHODS: Young male and female rats (approximately 3 months old) were assigned to either a CET group, which received a 20% ethanol drinking solution for 20 weeks, or a pair-fed control group, which received ad libitum tap water and a restricted diet for 20 weeks. After the CET groups were phased off the 20% alcohol solution, learning and memory abilities were examined by using matching-to-position and nonmatching-to-position tasks. Neuropathology was assessed in the frontal cortex and medial septal region. RESULTS: CET was shown to cause behavioral deficits. The behavioral dysfunction was sex, task, and process dependent; i.e., CET-female rats displayed a delay-dependent impairment on delayed matching-to-position, whereas CET-male rats displayed a delay-independent impairment on delayed nonmatching-to-position. CET resulted in a significant reduction in the frontal cortical (FR1) and collosal thickness, as well as a decrease in cells staining immunopositive for choline acetyltransferase in the medial septal region. However, relative to male rats exposed to CET, female rats did not show any accelerated neuropathology after CET. CONCLUSIONS: Chronic exposure to ethanol does result in both brain and behavior dysfunction in male and female rats. The results demonstrate that different cognitive processes are altered by chronic ethanol exposure in male and female rats. However, the neurobiological mechanisms responsible for these differences remain to be determined. PMID- 10798583 TI - Involvement of dopamine D2 autoreceptors in the ventral tegmental area on alcohol and saccharin intake of the alcohol-preferring P rat. AB - BACKGROUND: The ventral tegmental area (VTA) dopamine (DA) system is considered to be involved in mediating the actions of ethanol (EtOH). The objective of the present study was to examine the role of VTA DA D2 receptors in regulating EtOH intake of alcohol-preferring P rats. METHODS: EtOH (10% v/v) and saccharin (SACC, 0.0125% g/v) intake during 2 hr of limited access was assessed after microinjections of the D2 agonist quinpirole and the D2 antagonist sulpiride into the anterior VTA (AVTA) of female P rats. Both EtOH-SACC alternate-day-access conditions and daily availability of EtOH and SACC solutions to separate groups of subjects were used. A second D2 agonist, quinelorane, and coadministration of 2.0 microg sulpiride with 2.0 microg quinpirole were tested in animals given limited access to EtOH. Finally, the effects of quinpirole injected 2 mm dorsal to the VTA and within the posterior VTA (PVTA) were assessed under EtOH-SACC alternate-day-access conditions. RESULTS: Microinjections of 2.0-6.0 microg quinpirole into the AVTA dose dependently decreased EtOH intake 40-80% during the first 30 min of the limited access sessions but did not alter SACC intake. Injections of 2.0-4.0 microg quinelorane into the AVTA also reduced EtOH intake in the first 30 min. Administration of 0.5-2.0 microg sulpiride into the AVTA had no effect on either EtOH or SACC intakes but did attenuate the effects of quinpirole on reducing EtOH intake. Injections of 2.0-4.0 quinpirole 2 mm dorsal to the VTA did not alter EtOH or SACC intakes. Posterior VTA injections of quinpirole decreased EtOH and SACC intakes approximately 25-30% and 60-70%, respectively, in the first 30 min. None of the treatments altered intakes during the 30-120 min period. CONCLUSIONS: The data suggest that DA neuronal activity within the AVTA may be important for maintaining EtOH drinking in P rats, whereas DA neuronal activity within the PVTA may be involved in regulating general drinking and/or motivational behaviors. Overall, the results confirm the involvement of mesolimbic DA in EtOH self-administration and suggest that there is functional heterogeneity within the VTA regulating drinking behavior of the P rat. PMID- 10798585 TI - Carbohydrate-deficient transferrin, gamma-glutamyltransferase, and macrocytic volume as biomarkers of alcohol problems in women. AB - BACKGROUND: Early identification of alcohol problems in women is important. Differences in patterns of drinking and in biological function between genders, however, may present unique difficulties in biochemical screening. METHODS: Published alcohol-screening studies with female samples and use as biomarkers of carbohydrate-deficient transferrin (CDT), gamma-glutamyltransferase (GGT), and macrocytic volume were reviewed. RESULTS: A wide range of sensitivities and specificities of GGT and CDT have been reported, although, in general, the two markers seem approximately equal in accuracy. As in the case of males, use of them in combination substantially enhances sensitivity and little reduces specificity. Use of macrocytic volume improves the sensitivity of both GGT and CDT as alcohol-screening markers. CONCLUSIONS: GGT and CDT have moderate sensitivity in screening for alcohol problems in women. Use of them in concert offers further advantage. PMID- 10798584 TI - Nifedipine alleviates alterations in hippocampal kindling after repeated ethanol withdrawal. AB - BACKGROUND: Current clinical treatment of alcohol detoxification commonly includes pharmacotherapy to lessen the potential for seizures, especially in those patients undergoing repeated treatment. Basic research continues to study the alcohol withdrawal-related "kindling" of seizures both to understand the mechanisms involved and to identify alternative treatments. Ethanol withdrawal has been shown to result in the delay of electrical kindling at several brain sites, which suggests a long-lasting disruption of neuronal function. METHODS: This study focused on the participation of the L-type voltage-gated calcium channels in this process by the treatment of animals during withdrawal with nifedipine, an agent that blocks these channels. Animals were randomly assigned to ethanol (ethanol-exposed/ethanol-naive) and drug treatment (nifedipine/vehicle) groups. Subjects receiving ethanol were exposed to five cycles of 3 days' ethanol exposure, with each exposure cycle separated by a 1-day withdrawal period. Drug treatment was administered twice during each withdrawal period. Twenty days after completion of ethanol exposure, animals received daily kindling stimulations to hippocampal area CA3 until the kindling criterion was attained. RESULTS: Ethanol-exposed animals that received vehicle treatment during ethanol withdrawal required more daily stimulations to become fully kindled than did ethanol-naive controls. This delay in seizure development was most pronounced in the progression from focal seizure behaviors to more generalized seizures. Animals that received the same ethanol exposure but that were treated with nifedipine required significantly fewer stimulations than did ethanol-exposed animals that received vehicle. Ethanol-exposed/ nifedipine-treated animals did not differ from ethanol-naive controls that received vehicle or nifedipine. CONCLUSIONS: Alcohol withdrawal-related alterations in seizure-sensitive neural circuitry such as the hippocampus persist long after cessation of ethanol exposure. Furthermore, the L-type voltage-gated calcium channels are involved in this effect in that blockade of these channels during acute withdrawal alleviates alterations in seizure mechanisms on a long-term basis. PMID- 10798586 TI - Carbohydrate-deficient transferrin (CDT) and HDL cholesterol (HDL) are highly correlated in male alcohol dependent patients. AB - BACKGROUND: Serum levels of total HDL cholesterol (HDL) are reportedly influenced by recent alcohol intake. We examined the correlation between HDL cholesterol and widely used markers of excessive alcohol intake, such as carbohydrate-deficient transferrin (CDT), gamma-glutamyl-transferase (GGT), or mean corpuscular volume of erythrocytes (MCV), of which CDT is thought to be the most specific. METHODS: Several serological markers [i.e., CDT, GGT, aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), MCV, and HDL] were determined in 100 actively drinking male patients with alcohol dependence (DSM-IV) and in 27 non-alcohol dependent controls, according to routine procedures. Spearman's rank correlation coefficients were calculated. RESULTS: We found a highly significant positive correlation between HDL and CDT (r(s) = 0.55; p < 0.0005) in patients, but not in controls (r(s) = 0.13;p = 0.51). HDL was also positively correlated with GGT, ALAT, ASAT, and MCV only in patients. CONCLUSIONS: HDL cholesterol, as a widely determined parameter, may represent a useful routine marker for recent excessive alcohol intake. High HDL cholesterol levels should alert clinicians to investigate a patient's recent pattern of alcohol consumption. PMID- 10798587 TI - Personality, family history, and alcohol use among older men: the VA Normative Aging Study. AB - BACKGROUND: We examined personality traits (Sociability, Impulsivity, Neuroticism) as mediators of the effects of family history on alcohol outcomes. METHODS: A sample of 485 men reported on family history of alcohol problems in 1973, completed the Eysenck Personality Inventory in 1976, and responded to a survey on alcohol use in 1982. RESULTS: Using structural equation modeling, family history was found to have direct effects on number of drinks per day and on the number of alcohol problems, as well as indirect effects mediated through Neuroticism. There were no effects of Sociability or Impulsivity on either alcohol outcome. CONCLUSIONS: In this sample of older men, family history had both direct and indirect effects, and personality traits found to affect alcohol outcomes were different from those that have been found in younger men. PMID- 10798588 TI - Amelioration of ethanol-induced neurotoxicity in the neonatal rat central nervous system by antioxidant therapy. AB - BACKGROUND: The cerebellum of the neonatal rat is highly susceptible to ethanol, with profound loss of Purkinje cells resulting from even brief exposure during the first postnatal week. Developmental ethanol exposure previously has been shown to induce free radicals/oxidative stress processes and/or down-regulate protective antioxidants. In an earlier study, we found antioxidants protected against ethanol neurotoxicity in a tissue culture environment. The present study was designed to determine whether similar protection could be manifested in the intact animal. METHODS: Neonatal rats were administered a liquid diet via intragastric intubation on postnatal days 4 and 5 (P4-P5), the peak period of ethanol sensitivity in the developing cerebellum. The diet consisted of milk formula with 12% ethanol, the isocaloric substitution of maltose-dextrin for ethanol, or ethanol plus the antioxidant vitamin E. Unbiased three-dimensional counting was utilized to analyze Purkinje cell numbers and density within defined volumes from these animals on P5. RESULTS: These determinations revealed a substantial loss of Purkinje cells in the ethanol-treated animals compared to controls (approximately 30-44%), but this loss was prevented by the inclusion of vitamin E (601U/100 ml) in the diet. A lower concentration of the antioxidant (301U/100 ml) was not effective in this regard, however. CONCLUSIONS: These results suggest that ethanol-related cerebellar damage during this early postnatal period may be related to oxidative stress processes or the insufficiency of protective antioxidants. Thus, antioxidant treatment may represent a possible therapy for preventing or ameliorating the central nervous system (CNS) damage seen in the fetal alcohol syndrome. PMID- 10798589 TI - Expansions of CD8+CD28- and CD8+TcRVbeta5.2+ T cells in peripheral blood of heavy alcohol drinkers. AB - BACKGROUND: Despite heavy alcohol consumption, only a low percentage of heavy drinkers develop liver disease. Imbalances in T-cell subsets and iron metabolism parameters are common findings in heavy drinkers, yet the possible role played by discrete T-lymphocyte subsets under heavy alcohol consumption remains unclear. METHODS: To gain new insights into the possible role played by T lymphocytes during alcohol consumption, characterization of CD28 expression and TcR repertoire in peripheral blood CD4+ and CD8+ T cells by two and three-color flow cytometry was performed. A group of heavy alcohol drinkers (AHD, n = 71) and a group of age-matched controls (n = 81), both HLA-phenotyped and HFE-genotyped, constituted the groups under study. RESULTS: Marked expansions of CD28- T cells within the CD8+ but not the CD4+ T-cell pool were observed in AHD compared with controls. These CD8+CD28- expansions were paralleled by expansions of CD8+ T cells bearing specific TcR Valpha/beta chains, namely VP5.2. Moreover, AHD, but not controls, carrying the H63D mutation in the HFE gene showed significantly higher percentages of CD28- T cells within the CD8+ T-cell pool than AHD carrying the normal HFE gene. Finally, high numbers of CD8+CD28- T cells in AHD were associated with lower levels of the liver-related enzymes ALT and GGT. CONCLUSIONS: This study showed that under active ethanol consumption, expansions of discrete CD8+ T-cell subsets occur within the CD8+ T-cell pool, that molecules of the MHC-class I locus seem to influence the extent of the expansions, and that high numbers of CD8+CD28- T cells are associated with low levels of liver enzymes in AHD. PMID- 10798590 TI - Effects of acute, moderate ethanol consumption on human platelet aggregation in platelet-rich plasma and whole blood. AB - BACKGROUND: Platelet-inhibitory effects of alcohol potentially contribute to the reduced risk of coronary heart disease associated with moderate drinking. However, few studies have directly examined the effects of acute consumption of a moderate dose of alcohol on aggregation of platelets from healthy human subjects. In the present study, we examined those effects, with the use of multiple platelet agonists and two aggregation measurement techniques, as part of an ongoing series of studies that evaluate the actions of ethanol on platelet function. METHODS: Human subjects consumed alcohol at a dose equivalent to one drink (0.25 ml/kg) or two drinks (0.5 ml/kg). One hour after ingestion, anticoagulated blood was collected and agonist-induced aggregation of platelets was measured in both whole blood and in platelet-rich plasma. RESULTS: Inhibition of aggregation by ethanol consumption was observed in whole blood (measured as maximum change in impedance) and reached statistical significance (p < 0.05) in the 0.5 ml/kg alcohol group for two collagen concentrations (0.625 and 1.25 microg/ml) as well as for the highest concentration of adenosine diphosphate tested (p < 0.05). Inhibition was 15.4%, 22.6%, and 10.5%, respectively, for these three situations. In platelet-rich plasma, after consumption of 0.5 ml/kg ethanol, aggregation (measured as maximum change in optical density) in response to 1.25 microg/ml collagen was significantly inhibited (p < 0.05); no other significant inhibition was observed at either dose of alcohol in any other cases with platelet-rich plasma. In comparison of male and female subjects, there was a statistically significant difference (p < 0.05) in the degree of inhibition by ethanol consumption (0.5 ml/kg) of whole-blood platelet aggregation induced by collagen, whereby female aggregation was inhibited to a greater extent than male. CONCLUSIONS: This study shows that alcohol, at physiologically relevant doses, below those investigated in most previous human studies, has a dose-dependent inhibitory effect on platelet aggregation. Such an effect could potentially contribute to the beneficial effects of alcohol consumption against coronary artery disease. The inhibitory action is most readily measured with whole-blood platelet aggregometry, with the use of collagen as the agonist. This observation is consistent with the view that alcohol reduces platelet sensitivity to thrombotic stimuli by inhibition of arachidonic acid release and, therefore, subsequent thromboxane synthesis. PMID- 10798591 TI - Ethanol induces Fas/Apo [apoptosis]-1 mRNA and cell suicide in the developing cerebral cortex. AB - INTRODUCTION: Animal studies modeling fetal alcohol syndrome have demonstrated that developmental exposure to alcohol is associated with decreased brain weight and significant neuronal loss in multiple regions of the developing brain. Our previous data suggest that the Fas/Apo [apoptosis]-1 receptor is transiently expressed in the developing cerebral cortex during the peak period of naturally occurring apoptotic cell death and maximum sensitivity to alcohol. Therefore, we hypothesized that ethanol increases the expression of suicide receptors such as Fas/Apo-1 in the developing fetal cerebral cortex and leads to an upregulation or extension of the normal period of apoptosis and consequent disorganization of the neural circuitry. METHODS: Ethanol was administered in one of four doses (120, 320, 630, and 950 mg/dl) to organotypic explant cultures of the developing cerebral cortex established from postnatal day 2 rats and maintained for 6 days in vitro. The number of cells expressing Fas/Apo-1 receptor mRNA was counted. Apoptosis was measured by the use of two independent assays; a cell death enzyme linked immunosorbent assay for DNA fragmentation and flow cytometric analysis of Annexin-V binding to phosphatidylserine externalized to the outer leaflet of the plasma membrane. Necrosis was also estimated by two independent measures, the amount of lactate dehydrogenase released into culture medium and flow cytometric analysis of cells that were positive for both Annexin-V and propidium iodide. RESULTS: A significantly larger number of developing cortical cells expressed Fas/Apo-1 mRNA at the lower doses (120 and 320 mg/dl) than at the higher doses (630 and 950 mg/dl). Furthermore, ethanol induced apoptosis in a dose-related manner, with peak apoptosis observed at a dose of 630 mg/dl in the case of DNA fragmentation and at 630 and 950 mg/dl in the case of phosphatidylserine translocation to the outer leaflet of the plasma membrane. Ethanol did not induce necrosis at any of the administered doses of ethanol. CONCLUSIONS: Our data suggest that ethanol induces a susceptibility to apoptotic signals at low doses by upregulating the expression of mRNAs for cytotoxic receptors such as Fas/Apo-1 in the developing cerebral cortex. However, ethanol itself specifically induces apoptosis in the developing cerebral cortex only at higher doses. PMID- 10798592 TI - Formation of malondialdehyde adducts in livers of rats exposed to ethanol: role in ethanol-mediated inhibition of cytochrome c oxidase. AB - BACKGROUND: Previous studies in our laboratory demonstrated that short-term ethanol consumption by maternal rats increased the hepatic levels of 4 hydroxynonenal (HNE) in both the adult and the fetus. Additionally, HNE inhibited cytochrome c oxidase (COX) by forming adducts with the enzyme subunits. The present study examined modification of COX by another major aldehydic lipid peroxidation product, malondialdehyde (MDA), and its role in COX inhibition by ethanol. METHODS AND RESULTS: It is demonstrated in vitro that MDA inhibits the activity of purified COX while forming adducts with the enzyme. Compared with HNE, MDA is a more potent inhibitor of COX. Overnight incubation at room temperature caused an 80% decrease in COX activity by MDA versus a 67% decrease by HNE. MDA produced marked inhibition of COX activity at physiologically relevant concentrations, e.g., 43% inhibition at 10 microM. Although our previous studies documented that HNE formed adducts primarily with subunit IV of COX via histidine residues, the current report showed that MDA forms adducts with both subunit IV and subunit V via lysine residues. Furthermore, both aldehydes induce carbonyl formation in subunit IV. The in vivo role of MDA in the impairment of COX by ethanol is assessed in both adult and fetal liver after maternal ethanol consumption. CONCLUSIONS: The results showed that: (1) there are significant increases in MDA levels in liver homogenate as well as mitochondria in both adult and fetal livers after ethanol exposure; (2) these MDA levels are in the nanomole/mg protein range, in contrast to picomole/mg protein range of HNE in identical setting; and (3) ethanol-induced production of MDA is accompanied by enhanced formation of MDA adducts with COX. These findings suggest that MDA may play at least as equally an important role as HNE in ethanol-induced inhibition of COX. PMID- 10798593 TI - Ethanol and murine interleukin (IL)-12 production. AB - BACKGROUND: Interleukin (IL)-12 is a cytokine with protean effects against bacterial and intracellular pathogens. Induction of IL-12 at the time of infection has salient effects on elimination of various microbes. This work describes the effect of exposure to ethanol on lipopolysaccharide (LPS)-induced production of IL-12 in mice and whether ethanol-induced increases in IL-10 mediates these changes in IL-12 production. METHODS: BALB/c mice were pretreated with ethanol and then challenged with LPS either intravenously (iv) or intratracheally (it), and blood and lung production of IL-12 (p70 and p40 components) and serum IL-10 were assayed. Splenic and lung mRNA for IL-12 p35 and p40 components was determined. RESULTS: Ethanol pretreatment suppressed LPS induced IL-12 p70 and p40 protein production in blood and lung. In spleen and lung, p40 mRNA was induced to a greater extent than p35 mRNA, and there was greater suppression of p40 mRNA compared with p35 mRNA in ethanol-treated animals. Ethanol up-regulated the production of IL-10, and pretreatment of these animals with a polyclonal anti-IL-10 antibody resulted in significant increases in IL-12 p70 and p40 levels, but not completely to those of control animals. CONCLUSIONS: Ethanol suppresses the production of murine IL-12 in response to LPS in blood and lung, with both the p70 and the p40 components affected. This suppression is accompanied by reductions of p40 mRNA in both spleen and lung. IL 10 may play a role in ethanol-induced suppression of IL-12, as neutralization of IL-10 partially attenuated the suppression of IL-12. PMID- 10798594 TI - Decreased natural killer cell responses and altered interleukin-6 and interleukin 10 production in alcoholism: an interaction between alcohol dependence and African-American ethnicity. AB - BACKGROUND: Epidemiological data indicate that among alcoholics African-American men have higher rates of infections compared with white alcoholics. Alcohol is thought to reduce natural killer (NK) cell responses and to alter cellular immunity by changing the relative balance of Th1 versus Th2 cytokine response profiles. However, translation of these basic immunological observations into the clinical setting is limited, and what clinical data are available are contradictory, in part, because of heterogeneity within the alcoholic patient population. Whether ethnic characteristics contribute to variation in the immunological effects of alcohol is not known. METHODS: This study compared NK activity, interleukin (IL)-2-stimulated NK activity, and concanavalin A stimulated peripheral blood mononuclear cell production of Th1 (IL-12 and IL-2), Th2 (IL-10), and proinflammatory (IL-6) cytokines in 31 hospitalized chronic alcoholic patients and 31 age-matched controls who were stratified on the basis of ethnicity. RESULTS: NK-cell responses were significantly different across the four groups, and African-American alcoholics showed the lowest levels of NK activity (F = 9.5;p < 0.001) and IL-2-stimulated NK activity (F = 2.9; p < 0.05). Compared with the other three groups, African-American alcoholics also showed lower levels of IL-6 (F = 7.2;p < 0.01) and higher levels of IL-10 (F = 4.9;p < 0.05). Stimulated production of IL-2 and IL-12 were similar in the four groups. Regression analyses showed that alcohol dependence and ethnicity predicted NK activity, whereas the interaction between alcohol dependence and ethnicity predicted levels of IL-6 and IL-10. CONCLUSIONS: In summary, the immunological effects of alcohol dependence are modified by ethnicity status, consistent with the increased health risks found in African-American alcoholics. PMID- 10798595 TI - Metronidazole increases intracolonic but not peripheral blood acetaldehyde in chronic ethanol-treated rats. AB - BACKGROUND: Metronidazole leads to the overgrowth of aerobic flora in the large intestine by reducing the number of anaerobes. According to our previous studies, this shift may increase intracolonic bacterial acetaldehyde formation if ethanol is present. Metronidazole is also reported to cause disulfiram-like effects after alcohol intake, although the mechanism behind this is obscure. Therefore, the aim was to study the effect of long-term metronidazole and alcohol treatment on intracolonic acetaldehyde levels and to explore the possible role of intestinal bacteria in the metronidazole related disulfiram-like reaction. METHODS: A total of 32 rats were divided into four groups: controls (n = 6), controls receiving metronidazole (n = 6), ethanol group (n = 10), and ethanol and metronidazole group (n = 10). All rats were pair-fed with the liquid diet for 6-weeks, whereafter blood and intracolonic acetaldehyde levels and liver and colonic mucosal alcohol (ADH) and aldehyde dehydrogenase (ALDH) activities were analyzed. RESULTS: The rats receiving ethanol and metronidazole had five times higher intracolonic acetaldehyde levels than the rats receiving only ethanol (431.4 +/- 163.5 microM vs. 84.7 +/- 14.4 microM,p = 0.0035). In contrast, blood acetaldehyde levels were equal. Cecal cultures showed the increased growth of Enterobacteriaceae in the metronidazole groups. Metronidazole had no inhibitory effect on hepatic or colonic mucosal ADH and ALDH activities. CONCLUSIONS: The increase in intracolonic acetaldehyde after metronidazole treatment is probably due to the replacement of intestinal anaerobes by ADH-containing aerobes. Unlike disulfiram, metronidazole neither inhibits liver ALDH nor increases blood acetaldehyde. Thus, our findings suggested that the mechanism behind metronidazole related disulfiram-like reaction might be located in the gut flora instead of the liver. PMID- 10798596 TI - Assessing the performance of neural encoding models in the presence of noise. AB - An analytical method is introduced for evaluating the performance of neural encoding models. The method addresses a critical question that arises during the course of the development and validation of encoding models: is a given model near optimal in terms of its accuracy in predicting the stimulus-elicited responses of a neural system, or can the predictive accuracy be improved significantly by further model development? The evaluation method is based on a derivation of the minimum mean-square error between actual responses and modeled responses. It is formulated as a comparison between the mean-square error of the candidate model and the theoretical minimum mean-square error attainable through an optimal model for the system. However, no a priori information about the nature of the optimal model is required. The theoretically minimum error is determined solely from the coherence function between pairs of system responses to repeated presentations of the same dynamic stimulus. Thus, the performance of the candidate model is judged against the performance of an optimal model rather than against that of an arbitrarily assumed model. Using this method. we evaluated a linear model for neural encoding by mechanosensory cells in the cricket cercal system. At low stimulus intensities, the best-fit linear model of encoding by single cells was found to be nearly optimal, even though the coherence between stimulus-response pairs (a commonly used measure of system linearity) was low. In this low-stimulus-intensity regime, the mean square error of the linear model was on the order of the power of the cell responses. In contrast, at higher stimulus intensities the linear model was not an accurate representation of neural encoding. even though the stimulus-response coherence was substantially higher than in the low-intensity regime. PMID- 10798597 TI - Stochastic modeling of facilitated neurosecretion. AB - Two models of neurosecretion were evaluated in terms of their ability to predict the dependency of quantal content (m) on the frequency of repetitive stimulation of a lobster motoneuron. First, the hypothesis that neurosecretion is limited by a fixed number of release sites was tested by the fit of the distribution of m by uniform and nonuniform binomial statistics. The obtained release probabilities suggest that frequency facilitation can be due to activation of a group of sites with high release probabilities. However, the fit obtained using this model is not statistically significant due to a large number of fitting parameters. Second, the hypothesis that neurosecretion is limited by the rates of exchange between the releasable pool and the total store of quanta and that each stimulus enhances quantal mobilization was tested. Monte Carlo simulation was carried out in accordance with this model and reproduced the observed distribution of m with very few fitting parameters and therefore with a high level of significance (>0.1). This result demonstrates that mobilization of extra vesicles with each stimulus is a mechanism that allows a very accurate and parsimonious quantitative description of frequency facilitation. PMID- 10798598 TI - A simple mathematical model of second-messenger mediated slow excitatory postsynaptic potentials. AB - We have developed a novel and simple mathematical model of a slow excitatory postsynaptic potential (EPSP) based on an abstraction of the processes of activation, inactivation, and summation of a cAMP, protein kinase A (PKA) dependent second-messenger cascade. The model describes the activation of receptors, G-proteins, and production of cAMP as the first stage and uses first order, non-rate-limited kinetics. The second stage corresponds to the release of active, PKA catalytic subunit and can use first- or higher-order kinetics. The third stage represents simple phosphorylation of ion channels and is limited by the number of channels available. The decay of each stage is based on first order, mass-action kinetics. These equations and some variations were solved numerically and values of the parameters were determined by fitting to a variety of experimental data from myenteric neurons of the guinea-pig ileum. The model produced a slow EPSP with a nonlinear stimulus-response relationship that resulted from the underlying kinetics of the signaling cascade. This system of equations is suitable for incorporation into a large-scale computer simulation, and the methodology should be generalizable to other pathways. PMID- 10798600 TI - Integrating top-down and bottom-up sensory processing by somato-dendritic interactions. AB - The classical view of cortical information processing is that of a bottom-up process in a feedforward hierarchy. However, psychophysical, anatomical, and physiological evidence suggests that top-down effects play a crucial role in the processing of input stimuli. Not much is known about the neural mechanisms underlying these effects. Here we investigate a physiologically inspired model of two reciprocally connected cortical areas. Each area receives bottom-up as well as top-down information. This information is integrated by a mechanism that exploits recent findings on somato-dendritic interactions. (1) This results in a burst signal that is robust in the context of noise in bottom-up signals. (2) Investigating the influence of additional top-down information, priming-like effects on the processing of bottom-up input can be demonstrated. (3) In accordance with recent physiological findings, interareal coupling in low frequency ranges is characteristically enhanced by top-down mechanisms. The proposed scheme combines a qualitative influence of top-down directed signals on the temporal dynamics of neuronal activity with a limited effect on the mean firing rate of the targeted neurons. As it gives an account of the system properties on the cellular level, it is possible to derive several experimentally testable predictions. PMID- 10798599 TI - Computational modeling of orientation tuning dynamics in monkey primary visual cortex. AB - In the primate visual pathway, orientation tuning of neurons is first observed in the primary visual cortex. The LGN cells that comprise the thalamic input to V1 are not orientation tuned, but some V1 neurons are quite selective. Two main classes of theoretical models have been offered to explain orientation selectivity: feedforward models, in which inputs from spatially aligned LGN cells are summed together by one cortical neuron; and feedback models, in which an initial weak orientation bias due to convergent LGN input is sharpened and amplified by intracortical feedback. Recent data on the dynamics of orientation tuning, obtained by a cross-correlation technique, may help to distinguish between these classes of models. To test this possibility, we simulated the measurement of orientation tuning dynamics on various receptive field models, including a simple Hubel-Wiesel type feedforward model: a linear spatiotemporal filter followed by an integrate-and-fire spike generator. The computational study reveals that simple feedforward models may account for some aspects of the experimental data but fail to explain many salient features of orientation tuning dynamics in V1 cells. A simple feedback model of interacting cells is also considered. This model is successful in explaining the appearance of Mexican-hat orientation profiles, but other features of the data continue to be unexplained. PMID- 10798601 TI - Gene-marked autologous hematopoietic stem cell transplantation of autoimmune disease. AB - In phase I (safety) trials, we have demonstrated the feasibility of autologous hematopoietic stem cell transplantation (HSCT) for patients with autoimmune diseases. Although this review comments on results of our phase I trials, the focus is on phase II (efficacy) trials using gene-marked autologous stem cells. PMID- 10798602 TI - Stem cell transplantation in experimental models of autoimmune disease. AB - A review of the experiments with animal models of autoimmune disease (AID) that have provided the rationale for the present clinical investigations on the use of autologous stem cells for treating patients with severe refractory AID. The various types of AID in laboratory animals and the recognition of the key-role of hematopoietic stem cells (HSC) in AID are discussed. Two animal models were employed for translational research on autologous bone marrow transplantation (BMT): adjuvant arthritis (AA) as model for rheumatoid arthritis (RA) and experimental allergic encephalomyelitis (EAE) as model for multiple sclerosis (MS). The principal aspects of the treatment, i.e., conditioning agents and doses and T cell depletion of the autograft, were investigated in relation to remission induction and the incidence of relapses. PMID- 10798603 TI - Does hemopoietic stem cell transplantation have a role in treatment of severe rheumatoid arthritis? AB - Based on animal models and limited clinical experience, there is considerable interest in use of high-dose immunosuppression followed by hemopoietic stem cell transplantation as treatment for severe rheumatoid arthritis. Because of its relatively low treatment-related mortality and morbidity, autologous transplantation is a more attractive option than allogeneic transplantation for initial clinical trials, even though anecdotal reports suggest that allogeneic transplantation has a greater likelihood of bringing about long-term disease control. The approach remains experimental with many unanswered questions such as the value and safety of high-dose therapy without transplantation, the need for T cell purging, the possible deleterious effects of post-transplant hemopoietic growth factors and the potential of "mini" allogeneic transplantation (a process whereby intense immunosuppression is combined with less intense myelosuppression). To achieve quick progress it is essential that clinical trials be carefully designed with all cases being reported to the Autoimmune Disease Stem Cell Project Database. PMID- 10798604 TI - Autologous stem cell transplantation in progressive multiple sclerosis--an interim analysis of efficacy. AB - Based on the good results of experimental transplantation in animal models of multiple sclerosis and of other autoimmune diseases, we have treated 24 patients suffering from chronic progressive multiple sclerosis with high-dose chemotherapy (BEAM regimen) followed by autologous blood stem cell rescue and antithymocyte globulin. Blood stem cells were mobilised with cyclophosphamide at 4g/m2 and G- (or GM-) CSF. In 9 cases, additional CD34+ cell-selection of the graft was performed. Here we update previously published results of this novel treatment, mainly with regard to clinical efficacy, as the median follow-up time has reached 40 months (range, 21-51). Infections were the principal toxicity early after the procedure, with death of a patient from aspergillosis 65 days post stem cell infusion. No serious late events occurred apart from a case of autoimmune thyroiditis that developed 11 months after transplant in a patient who had received a CD34+ cell-depleted graft. Mild and transient neurotoxicity was observed in 10 patients (42%), most probably associated with fever and infections. Eighteen patients (18/23; 78%) responded to the treatment, i.e., they were improved or stabilized, while five patients progressed, of which 4 had primary progressive disease. Of those improved or stabilised (18), 9 patients have maintained stable condition whereas 9 developed relapses or they slowly resumed progression, although their disability scores have not gotten worse than they were before transplantation. The probability of progression-free survival (compared to entry status) at 3 years is 92% for patients with secondary progressive disease and 39% for the primary progressive type. CD34+ cell selection did not seem to yield better results except for a delay in progression or in relapse after transplantation. These results appear better than those achieved by any other treatment of progressive multiple sclerosis, including beta interferon, but they need to be confirmed by other open or controlled studies in view of the well-known difficulty of judging objectively the effect of a treatment in patients with this disease. PMID- 10798605 TI - Intense immune suppression for systemic lupus--the role of hematopoietic stem cells. AB - The treatment of severe autoimmune diseases has been recently revitalized by the introduction of intense immune suppression with immune ablative intent followed by three different procedures. These are allogeneic hematopoietic stem cell transplantation (HSCT), autologous HSCT (using either marrow or peripheral blood), and intense immune suppression without stem cell support. Current trials suggest that high dose immune suppressive therapy with or without autologous hematopoietic stem cell support can induce remission of previously refractory disease. Follow-up is too brief to determine if intense immune suppression, and more specifically autologous HSCT, will ultimately cure SLE. It is conceivable that an allogeneic source of stem cells from a normal donor (e.g. HLA matched sibling) will be required to achieve a cure. It is also possible that autologous HSCT, even if not curative, may prolong the life of patients with otherwise high risk features. In carefully selected patients, the potential benefits of this procedure may outweigh the risks. PMID- 10798606 TI - Prospects of stem cell transplantation in autoimmune diseases. AB - Severe autoimmune diseases are not cured by current conventional treatments. Preclinical studies of treatments requiring stem cell transplantation have shown sustained remissions in some models of autoimmune diseases. Both autologous stem cell transplantation after high-dose immunoablative therapy and allogeneic stem cell transplantation have a profound effect on the recipient's immunological system. Pilot clinical studies of autologous stem cell transplantation are being conducted. Sustained remissions will require the induction of tolerance to autoantigens during recovery. In a group of patients highly selected to minimize transplant-related mortality, allogeneic stem cell transplantation should be tested to determine if elimination of the autoreactive immunological system will achieve sustained remissions. These are promising therapies which will require long-term follow-up of patients to evaluate efficacy. PMID- 10798607 TI - Phagocytic killing and antibody response during the first year after tetravalent meningococcal vaccine in complement-deficient and in normal individuals. AB - Seven individuals with late complement component (LCC) deficiency and seven control subjects were vaccinated with tetravalent meningococcal vaccine. The response to vaccination was evaluated by measuring the antibody titer and the phagocyte killing of the bacteria, before, 5-7 weeks, and 12-14 months after vaccination. Prior to vaccination, no phagocytic killing and a low titer of antibody was found in the LCC-deficient group and a low killing (mean of 40-58%, according to the serogroup) in normal controls. The phagocytic killing increased significantly 5-7 weeks after vaccination. However, while in normal controls the phagocytic killing was close to 100% after 5-7 weeks and decreased only slightly during the first year, the mean killing of the various meningococcal subgroups in LCC-deficient individuals was 70-89% and dropped to only 53-71% one year after vaccination. Six weeks after vaccination the mean antimeningococcal antibody titer increased similarly in the sera of LCC-deficient patients and controls. One year after vaccination the controls maintained the high concentration, while the LCC-deficient patients had tendency toward a decrease. In addition, the interpersonal variability of the antibody concentration, both in LCC-deficient individuals and in normal controls, was much higher than the phagocytic killing, with only a very mild increase in some individuals. Thus, it is possible that in spite of adequate increase of antimeningococcal antibody titer after vaccination of LCC-deficient individuals their immunity against the bacteria may not be optimal. Our data show also that phagocytic killing of meningococci is probably a more consistent assay than antibody titer levels for antimeningococcal immunity, especially in LCC-deficient patients. PMID- 10798609 TI - Lymphoproliferative responses to mitogens and prepared antigens of M. avium complex in patients with HIV infection. AB - Published reports have demonstrated that antigens of Mycobacterium avium complex (MAC) can suppress the normal response to mitogens in lymphoproliferation assays. We therefore studied the lymphoproliferative (LP) function of PBMC from 55 HIV infected patients and 16 controls in response to mitogens with and without MAC antigen. As expected, LP responses decline with progressive decline in CD4 count; MAC antigen in combination with PHA further suppresses that response in a dose dependent manner. More relevant were the LP responses in those with CD4 counts less than 100. All patients with MAC disease had poor responses (stimulation index, SI < 10) to PHA or anti-CD3 with or without MAC antigen. Those who did not have nor subsequently developed MAC were both good (SI > 10) or poor responders (SI < 10). The suppressive effect of MAC on lymphocyte function may serve as a weak virulence factor which is only relevant in severely immunocompromised HIV patients. PMID- 10798608 TI - Relationship between interferon-gamma, interleukin-10, and interleukin-12 production in chronic hepatitis C and in vitro effects of interferon-alpha. AB - In the current study, increased interferon (IFN)-gamma, interleukin (IL)-10, and IL-12 p40 serum levels were observed in patients with chronic hepatitis C (CHC) compared to controls. Patients also displayed an increased spontaneous IFN-gamma release but a deficient peripheral blood mononuclear cells (PBMC) IFN-gamma production following stimulation with Staphylococcus aureus Cowan I strain (SAC). No difference was found with reference to spontaneous or phytohaemagglutinin (PHA)-induced IL-10 release between patients and controls, whereas a higher IL-12 p70 and IL-12 p40 secretion triggered by SAC was observed in patients. Moreover, IL-12 p40/p70 ratio following SAC stimulation was higher in patients compared to controls and a negative correlation was found between this ratio and IFN-gamma amounts. Recombinant IL-12 (rIL-12) as well as neutralizing anti-IL-10 monoclonal antibodies (mAbs) were able to restore the compromised IFN-gamma production. Of note, anti-IL-10 supplementation induced a lower IL-12 p40/p70 ratio in HCV subjects as compared to controls. Finally, IFN-alpha upregulated in vitro IFN gamma, IL-10, and IL-12 p70 release but not IL-12 p40 secretion, this giving rise to a normalization of IL-12 p40/p70 ratio. The data suggest the occurrence of an enhanced responsiveness to IL-10 modulating effects, likely mediated by an imbalance of IL-12 p40/p70 ratio, in chronic HCV infection. Cytokine balance restoration might thus contribute to achieve therapeutical results in chronic hepatitis C. PMID- 10798610 TI - Distal proctocolitis and n-3 polyunsaturated fatty acids (n-3 PUFAs): the mucosal effect in situ. AB - It has been postulated that patients with ulcerative colitis (UC) have altered reactivity of gut-associated lymphoid tissue. In such cases there is intense infiltration of the mucosa with immune competent cells and associated tissue damage. We have shown previously that the dietary supplementation with the n-3 polyunsaturated fatty acids (n-3 PUFAs), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) results in significant systemic immune suppression. The aim of this study, therefore, was to evaluate the in situ effect of n-3 PUFAs on distal proctocolitis. Each patient received either fish oil extract (EPA 3.2 g, DHA 2.4 g) (n = 9) or sunflower oil (n = 9) daily in a double blind manner for six months. Monthly assessment included: (1) disease activity using clinical, sigmoidoscopic, and histological scores and (2) immunohistochemical analysis (immunoglobulins, CD profiles) of rectal biopsy specimens (before and after six months supplementation) using monoclonal antibodies and quantitative computer assisted video image analysis. Prior to receiving supplementation, patients with proctocolitis (n = 18) showed significantly higher numbers of cells expressing CD3 (pan T cells) and HLA-DR and IgM containing cells compared with non-colitic controls (n = 8). Six months supplementation with n-3 PUFAs resulted in significant reduction in the number of cells expressing CD3 and HLA and the percentage of cells containing IgM. There was no significant change in the CD20 nor the percentage of IgG or IgA containing cells in either group of patients with procto-colitis. In patients receiving n-3 PUFA supplementation, there was improvement in the disease activity and histological scores, compared with pretreatment evaluation. This study has demonstrated both evidence of suppression of in situ immune reactivity and concurrent reduction in disease activity in patients with proctocolitis receiving n-3 PUFA supplementation. This may have important implication for therapy in patients with ulcerative colitis. PMID- 10798611 TI - IgG anti-IgA subclasses in common variable immunodeficiency and association with severe adverse reactions to intravenous immunoglobulin therapy. AB - The current therapy for common variable immunodeficiency is based on the administration of intravenous immunoglobulin preparations which may cause severe adverse reactions. Some reports have associated these reactions with IgG anti-IgA antibodies, although this is not yet clear. We analyzed 20 sera from common variable immunodeficiency patients by an enzyme immunoassay to detect IgG anti IgA and determine its subclass profile. Five patients presented high levels of these antibodies, all of them had IgG1, two had IgG2 and IgG4 and one had IgG3. Three of these five patients were receiving non IgA depleted intravenous immunoglobulin and had no severe adverse reactions. One patient had persisted with similar high levels of IgG anti-IgA during three years. Therefore, the IgG anti-IgA antibodies, regardless to their subclass profile in the common variable immunodeficiency patients sera do not seem to be associated with severe adverse reactions to intravenous immunoglobulins. PMID- 10798613 TI - Specific association of a small protein with the telomeric DNA-protein complex during the onset of leaf senescence in Arabidopsis thaliana. AB - Telomeres and their changes in length throughout the life span of cells have been intensively investigated in different organisms. Telomere length is assumed to control replicative senescence in mammalian cells. However, only very few data are available on the developmental dynamics of plant telomeres. Here, changes of telomere length and DNA-protein structure of Arabidopsis thaliana telomeres were analysed in different stages of development, with the main focus resting on the transition from pre-senescent to senescent leaves. The lengths of the telomeres, ranging from ca. 2.0 to 6.5 kb, do not significantly change during plant development indicating that telomere length is not involved in differentiation and replicative senescence nor in post-mitotic senescence of A. thaliana. In dedifferentiated cultured cells a slight increase in length can be determined. The nucleoprotein structure of the telomeric DNA was investigated by gel mobility shift assays, with synthetic oligonucleotides and nuclear protein extracts derived from four defined stages of post-mitotic leaf senescence. In all four stages, a highly salt-resistant DNA-protein complex was formed with the double stranded as well as with the single-stranded G-rich telomeric DNA. An additional DNA-protein complex was identified in nuclear protein extracts isolated from plants in the transition stage from pre-senescence to senescence. The protein components of the DNA-protein complexes were analysed on native PAGE and SDS-PAGE gels. A protein of 67 kDa (ATBP1) bound to the telomeric DNA in all developmental stages. An additional protein of merely 22 kDa (ATBP2) was associated via protein protein interaction with ATBP to form a higher-order complex exclusively during the onset of senescence. DNA interaction of this higher-order protein complex seems to be restricted to double-stranded telomeric DNA. The defined period of ATBP1/ATBP2 complex formation with the telomeric DNA probably indicates that ATBP2 is involved in the onset of post-mitotic leaf senescence by either disturbing an established or establishing an additional function exhibited by the telomeres in the interphase nuclei. PMID- 10798612 TI - Plant homologue of flap endonuclease-1: molecular cloning, characterization, and evidence of expression in meristematic tissues. AB - Flap endonuclease-1 (FEN-1) is an important enzyme involved in DNA replication and repair. We isolated a 1.4 kb cDNA from rice (Oryza sativa), termed OsFEN-1, encoding a protein which shows homology with the eukaryotic FEN-1 proteins. OsFEN 1 protein was overexpressed in Escherichia coli and purified to near homogeneity. DNA cleavage analysis using different branched DNA structures indicated that OsFEN-1 protein possesses both 5'-flap endonuclease and 5' to 3' double-stranded DNA exonuclease activities. OsFEN-1 protein incises a 5'-flap and 5'-pseudo Y structure one base 3' of the branched point in the duplex region. The enzymatic properties indicated that we succeeded in obtaining the gene and the protein of a plant counterpart of FEN-1. OsFEN-1 transcripts were expressed strongly in proliferating tissues such as root tips and young leaves that contain root apical meristem and marginal meristem, respectively. No expression was detected in mature leaves although the leaves were exposed to UV. We analyzed the spatial distribution pattern of OsFEN-1 transcripts by in situ hybridization. In the shoot apex, OsFEN-1 mRNA was abundant in the shoot apical meristem, tiller bud, leaf primordia, ligule primordia and marginal meristem of young leaves. In the roots, the transcript accumulated to high levels in the root apical meristem. Our results indicate that OsFEN-1 is expressed in tissues rich in proliferating cells, and its expression may be required for cell growth and organ formation. PMID- 10798614 TI - Chlorophyll breakdown in Chlorella protothecoides: characterization of degreening and cloning of degreening-related genes. AB - Chlorella protothecoides cultures grown in a nitrogen-free bleaching medium (BM N) in the dark rapidly degraded chlorophyll (Chl) to red catabolites. This degreening process was investigated under different growth conditions. Supply of nitrogen to the culture medium (BM+N) inhibited bleaching and the synthesis of catabolites as did the addition to BM-N of cycloheximide or a chelator, 2,2' bipyridyl. In contrast, chloramphenicol or the protease inhibitor E64 had no effect. During bleaching, Chl breakdown was accompanied by the degradation of cellular proteins such as light-harvesting complex II, cytochrome f and protochlorophyllide oxido-reductase. During growth in BM-N, protease activity increased and proteins immunologically detectable with an antibody against a senescence-enhanced cysteine protease accumulated. cDNAs from BM-N and BM+N cells were used for differential and subtractive screening to isolate cDNAs representing genes with degreening-enhanced expression (dee) in C. protothecoides. Several different dees were identified with different patterns of expression during Chlorella growth but which were all expressed at higher levels during bleaching. Among these, dee4 was most abundant and its expression was exclusive in BM-N cultures. Analysis of the dee sequences showed that they encode different proteins including a novel amino acid carrier (dee4), ferritin, ATP dependent citrate lyase, a Ca2+-binding protein, MO25, ubiquinone-cytochrome c reductase and several new proteins. PMID- 10798615 TI - Glucose modulates the abscisic acid-inducible Rab16A gene in cereal embryos. AB - Glucose effects on the expression of the abscisic acid-inducible Rab16A gene were examined in rice and barley embryos. Glucose feeding to rice embryos negatively affects the endogenous abscisic acid content and represses the promoter activity of the Rab16A gene. Glucose repression of the Rab16A gene takes place both at a transcriptional and a post-transcriptional level. Modulation of the abscisic acid content in rice embryos triggered by glucose did not directly influence the expression of the rice alpha-amylase gene RAmy3D, which is known to be under glucose control. The possible interaction between the glucose and abscisic acid signaling pathway is discussed. PMID- 10798616 TI - Tissue-specific expression of Pa18, a putative lipid transfer protein gene, during embryo development in Norway spruce (Picea abies). AB - A full-length Picea abies cDNA clone Pa18, encoding a protein with the characteristics of plant lipid transfer proteins, has been isolated and characterized. The size of the deduced 173 amino acid (aa) long protein is around 18 kDa. The first 100-120 aa show similarity to angiosperm lipid transfer proteins in amino acid sequence as well as in predicted secondary structure. The Pa18 gene is constitutively expressed in embryogenic cultures of Picea abies representing different stages of development as well as in non-embryogenic callus and seedlings. The Pa18 gene product has an antimicrobial activity. In situ hybridization showed that the Pa18 gene is equally expressed in all embryonic cells of proliferating embryogenic cultures but during embryo maturation the expression of the gene in maturing and mature somatic as well as in mature zygotic embryos is stronger in the outer cell layer than in other tissues. Southern blot analysis at different stringencies was consistent with a single gene with one or two copies rather than a gene family. Twenty independent transgenic sublines over- and under-expressing the Pa18 gene under the Zea mays ubiquitin promoter were established. There was a high yield of mature somatic embryos with a smooth surface only in untransformed, control cultures. Irrespective of the expression level of Pa18, the somatic embryos started to mature when given a maturation treatment. However, in the transgenic sublines, the outer cells in the maturing embryos frequently became elongated and vacuolated instead of remaining small and uniform. One explanation for this was that the expression of Pa18 was not restricted to the outer cell layer in transformed sublines. Angiosperms and gymnosperms separated about 300 million years ago and the embryo genesis is different in the two groups. The outer cell layer (protoderm), the first tissue to differentiate, is less clearly delineated in gymnosperms. For normal embryo development in angiosperms, expression of the LTP gene must be restricted to the protodermal cells. In this work we show that the expression of the Pa18 gene must be restricted to the putative protodermal cells of the gymnosperm. PMID- 10798618 TI - Isolation and characterization of the wheat prolyl isomerase FK506-binding protein (FKBP) 73 promoter. AB - The wheat FK506-binding protein (FKBP) 73 is a member of the peptidyl prolyl cis trans isomerase gene family, which catalyses the interconversion between the cis and trans forms of the peptide bond preceding proline residues in proteins. A 3.5 kb sequence 5' upstream of the ATG codon of the wheat FKBP73 was isolated from a wheat genomic library, and characterized by deletion analysis and transient expression in wheat embryos. The 1517 bp fragment is referred to as the full promoter due to the maximal activity of the fused luciferase reporter gene. Sequence analysis revealed the presence of three abscisic acid (ABA)-responsive elements (ABREs) proximal to coupling elements (CE1-like), a putative lectin box, two putative binding sites for the myb transcription factor and a 36 bp fragment which exhibits 100% identity to the pSau3A9 clone located in the centromeric region of wheat chromosomes. In a transient expression assay the promoter preserved the tissue specificity described in vivo, namely it is expressed only in germinating embryos and young shoots. The promoter was induced 1.9-fold by ABA, the minimal promoter was designated at -221 and the TATA box located at 137. The inducibility by ABA and the expression during germination may indicate that FKBP73 belongs to the group of genes induced by ABA upon germination. PMID- 10798617 TI - Secreted proteins of tobacco cultured BY2 cells: identification of a new member of pathogenesis-related proteins. AB - Cultured cells of tobacco BY2 secrete more than 100 proteins into culture medium. Six major proteins were purified, and partial protein sequences were determined. Five of them were found to be similar to an ascorbic acid oxidase, three peroxidase isozymes and a beta-1,3-exoglucanase, respectively. A cDNA clone encoding the remaining polypeptide, whose amino acid sequence showed no similarity with earlier reported proteins, was isolated. It encoded a putative 27 kDa protein of 242 amino acids with resemblance to WCI-5, a wheat protein induced by benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) which activates genes involved in systemic acquired resistance. Transcripts of this clone accumulated upon tobacco mosaic virus infection, mechanical wounding and drought treatment, an induction profile that satisfies the definition of pathogenesis-related (PR) proteins by van Loon et al. (Plant Mol. Biol. Rep. 12 (1994) 245). No similar PR proteins have so far been reported, and therefore our newly designated NtPRp27 points to the existence of a novel PR protein family in tobacco plants. PMID- 10798619 TI - Characterization of the potato MADS-box gene STMADS16 and expression analysis in tobacco transgenic plants. AB - A new MADS-box gene, STMADS16, has been cloned in Solanum tuberosum L. that is expressed in all vegetative tissues of the plant, mainly in the stem, but not in flower organs. STMADS16 expression is established early during vegetative development and is not regulated by light. Sequence similarity besides the spatial and temporal expression patterns allow to define a novel MADS-box subfamily comprising STMADS16 and the gene STMADS11. Expression of the STMADS16 sense cDNA under the control of the 35S cauliflower mosaic virus promoter modifies the inflorescence structure by increasing both internode length and flower proliferation of the inflorescence meristems, and confers vegetative features to the flower. Moreover, STMADS16 ectopic expression overcomes the increase in flowering time and node number produced under short-day photoperiod, while the flowering time is not affected in long-day conditions. These results are discussed in terms of a possible role for STMADS16 in promoting vegetative development. PMID- 10798620 TI - Localization of AtROP4 and AtROP6 and interaction with the guanine nucleotide dissociation inhibitor AtRhoGDI1 from Arabidopsis. AB - The small GTPases of the Rho family play a key role in actin cytoskeletal organization. In plants, a novel Rho subfamily, called ROP (Rho of plants), has been found. In Arabidopsis, 12 ROP GTPases have been identified which differ mainly at their C-termini. To test the localization of two members of this subfamily (AtROP4 and AtROP6), we have generated translational fusions with the green fluorescent protein (GFP). Microscopic analysis of transiently transfected BY2 cells revealed a predominant localization of AtROP4 in the perinuclear region, while AtROP6 was localized almost exclusively to the plasma membrane. Swapping of the AtROP4 and AtROP6 C-termini produced a change in localization. As RhoGDIs are known to bind to the C-terminus of GTPases of the Rho family, we searched for Arabidopsis RhoGDI genes. We identified the AtRhoGDI1 gene and mapped it to chromosome 3. AtRhoGDI1 encodes a 22.5 kDa protein which contains highly conserved amino acids in the isoprene binding pocket and exhibits 29% to 37% similarity to known mammalian RhoGDI homologues. The AtRhoGDI1 gene was expressed in all tissues studied. Using the yeast two-hybrid system, we showed specific interaction of AtRhoGDI1 with both AtROP4 and AtROP6 as well as with their GTP-locked mutants, but not with a GTPase of the RAB family. Recombinant GST-AtRhoGDI1 could bind GFP-AtROP4 from transgenic tobacco BY2 cell extracts, confirming the interaction observed with the two-hybrid system. PMID- 10798621 TI - Male-sterile tobacco displays abnormal mitochondrial atp1 transcript accumulation and reduced floral ATP/ADP ratio. AB - Alloplasmic male sterility is commonly obtained in Nicotiana by combining the nucleus from N. tabacum with the cytoplasm from other Nicotiana species. Besides being male-sterile, most of these cultivars also display changes in floral organ structure. Flowers from male-sterile plants containing the nucleus from N. tabacum combined with N. repanda cytoplasm develop stamen with shortened filaments and shrivelled anthers capped with stigmatoids. Male fertility and normal floral development can be restored by introduction of a restorer chromosome fragment from the cytoplasmic donor N. repanda into the N. tabacum nucleus. A novel reading frame, orf274, located upstream of atp1 in the mitochondrial genomes of both N. tabacum and N. repanda, as well as in the male sterile and fertility-restored plants was identified. Co-transcripts of orf274 and atp1 were detected by RT-PCR in all four cultivars, but these transcripts accumulate to levels detectable by northern hybridization only in male-sterile plants. These co-transcripts neither generate detectable levels of an ORF274 polypeptide nor lead to altered expression of the ATP synthase subunit alpha. Measurement of ATP and ADP steady-state levels, however, revealed that the ATP/ADP ratio is significantly lower in young floral buds of male-sterile plants than in fertile plants. PMID- 10798622 TI - Comparative in vitro percutaneous absorption of nonylphenol and nonylphenol ethoxylates (NPE-4 and NPE-9) through human, porcine and rat skin. AB - The purpose of this study was to assess the percutaneous absorption of nonylphenol (NP) and the nonylphenol ethoxylates, NPE-4 and NPE-9, in human, porcine and rat skin. In vitro studies with the NPEs were conducted for 8 h in flowthrough diffusion cells using topical solutions of 0.1, 1.0 and 10% in PEG 400 or 1% in water (NPE-9 only). NP absorption was assessed as a 1% solution in PEG-400. All compounds were 14C ring-labeled and radioactivity in perfusate was monitored over time. Skin deposition was measured at the termination of the experiment. Absorption into perfusate and total penetration (compound absorbed plus compound sequestered in skin) were calculated. Absorption of NPE-4, NPE-9 and NP was similar across all species at less than 1% of the applied dose over 8 h. Penetration was generally below 5% of applied dose, the majority located in the stratum corneum. In all species and for both NPEs, the fraction of dose absorbed was highest for the lowest applied dose. Absorptions expressed as actual mass absorbed over 8 h were similar (approximately 0.3 microg/cm2) across all concentrations. Penetration, but not absorption, was greater from a water vehicle compared to a PEG-400 vehicle, particularly in rat skin. These studies suggest that NP, NPE-4 and NPE-9 were minimally absorbed across skin from all three species. Fractional absorption was concentration-dependent, making the actual absorbed flux constant across all doses. PMID- 10798623 TI - The activity of erythrocyte enzymes and basic indices of peripheral blood erythrocytes from workers chronically exposed to mercury vapours. AB - The influence of occupational exposure to mercury vapours on the activity of the red cell enzymes [glucose-6-phosphate dehydrogenase (G-6PD), acetylcholinesterase (AChE), glutathione reductase (GR) and superoxide dismutase (SOD)], as well as on peripheral blood indices [erythrocyte number (RBC), HCT, Hb, MCHC] and on serum concentrations of iron, ferritin, transferrin and total iron binding capacity (TIBC), was assessed. Studies were carried out on 46 men aged between 21 and 56 years (X = 39 +/- 10.4) exposed to mercury vapours during their work from 7 months to 32 years (= 14.7 +/- 10.8). The control group consisted of 35 healthy workers aged between 20 and 54 years (X = 33.6 +/- 9.8) not exposed to chemical nor physical agents. In both groups studied, there were 50% and 34.3% smokers, respectively. The activity of studied red cell enzymes--G-6PD, AChE, GR and SOD- was estimated according to the colorimetric methods described by Beutler and expressed as international units per gram of hemoglobin (IU g Hb(-1)). Peripheral blood cell parameters were determined using an automatic cell counter. The concentration of serum iron and TIBC was determined using colorimetric methods (Beckman), while that of ferritin and transferrin by nephelometric methods. The time-weighted average (TWA) of mercury concentration in the air determined before the study was 0.0028 mg m(-3). Statistical analysis of the data was performed using either the Cochran and Cox C-test or the Student's t-test. The medium mercury concentration in the urine was 77.44 +/- 48.15 microg l(-1). In the group exposed to mercury vapours, a significant decrease was found in G-6PD activity (23.9%, P<0.001), GR (18.8%, P<0.001), and SOD (5%, P<0.001) with a concomitant increase in AChE activity (35.9%, P<0.001) was found. Moreover, a statistically significant increase occurred in HCT and RBC, and a decrease in MCV and MCHC as well as increases of ferritin (130.9%, P<0.001), transferrin (118.4%, P<0.001) and TIBC (11.2%, P<0.05). Our results indicate that long-term exposure to mercury vapours induces changes in the activity of red cell enzymes--G-6PD, AChE, GR and SOD--and may also influence other important hematological parameters of the peripheral blood. PMID- 10798624 TI - Breast-feeding exposure of infants to selected pesticides: a public health viewpoint. AB - In this paper, we provide an overview of the public health implications of exposure to some pesticides via breast milk and provide health-based guidance. The presence of organochlorine pesticides in breast milk has been documented in many studies around the world. Included in our review are aldrin/dieldrin, chlordane, 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT)/1,1-dichloro-2,2 bis(p-chlorophenyl)ethylene (DDE), endrin, hexachlorobenzene (HCB), and hexachlorocyclohexane (HCH). Toxicological and environmental data on these chemicals are compiled in toxicological profiles published by the Agency for Toxic Substances and Disease Registry (ATSDR). Based on the data, ATSDR derives chemical-specific minimal risk levels (MRLs) that assist in evaluating public health risks associated with exposure. MRLs are health-based guidance values designed to protect the most sensitive populations, including breast-fed infants. We compare MRLs and projected intakes from the breast milk for the listed pesticides, explore the possibilities of toxicological interactions, and provide health-based recommendations. PMID- 10798625 TI - Effects of short-term JP-8 jet fuel exposure on cell-mediated immunity. AB - The U.S. Air Force has implemented the widespread use of JP-8 jet fuel in its operations, although a thorough understanding of its potential effects upon exposed personnel is unclear. Exposure to environmental toxicants such as JP-8 may have significant effects on host physiology. Jet fuel exposure has been shown to cause human liver dysfunction, abnormal electroencephalograms, shortened attention spans, and decreased sensorimotor speed. Previous studies have shown that short-term, low-concentration JP-8 exposure had significant effects on the immune system; e.g., decreased viable immune cell numbers, decreased immune organ weights, and loss of immune function that persisted for extended periods of time (i.e., up to 4 weeks post-exposure). In the current study, an in-depth analysis of the effects of JP-8 exposure on cellular immunity was performed. Short-term (7 days, 1 h/day), low-concentration (1000 mg/m3) exposures were conducted in mice, and T cell and natural killer (NK) cell functions were analyzed 24 h after the last exposure. The exposure regimen was found to almost completely ablate NK cell function, as well as significantly suppress the generation of lymphokine activated killer (LAK) cell activity. Furthermore, JP-8 exposure suppressed the generation of cytotoxic T lymphocyte (CTL) cells from precursor T cells, and inhibited helper T cell activity. These findings demonstrate that JP-8 jet fuel exposure has significant detrimental effects on immune functions of exposed individuals. JP-8 jet fuel should be considered a potential and significant immunotoxicant. Chronic exposure to JP-8 may have serious implications to the long-term health of exposed individuals. PMID- 10798626 TI - Imaging in thoracic trauma: the trauma surgeon's perspective. AB - Thoracic trauma is a common cause of significant disability and mortality. Most thoracic injury in developed countries results from motor vehicle crashes (MVC). Imaging of patients with thoracic trauma must be accurate and timely to avoid preventable death. Trauma surgeons prioritize imaging options based on the patient's hemodynamic status, associated injuries, and age. The screening test for the detection of life-threatening thoracic injury is the supine anteroposterior (AP) chest radiograph. Rib fractures are a marker for serious associated injuries, including abdominal injuries. Rib fractures are especially ominous in children and the elderly. Thoracic aortic injury is associated with high-speed mechanisms of injury and can occur in the absence of radiographic signs. Chest computed tomography (CT) can be used as a screening and diagnostic tool for suspected aortic injury. Aortography is reserved for patients with high suspicion of aortic injury or for confirmation of CT scan diagnosis. PMID- 10798627 TI - Radiographic and CT findings in blunt chest trauma. AB - Injuries of the thorax are a major cause of morbidity and mortality in blunt trauma patients. Radiologic imaging plays an important role in the workup of the patient with thoracic trauma. The chest radiograph is the initial imaging study obtained, but computed tomography (CT) is now used frequently in the evaluation of chest trauma. The primary role of chest CT has been to assess for aortic injuries, but CT has been shown to be useful for the evaluation of pulmonary, airway, skeletal, and diaphragmatic injuries as well. Magnetic resonance imaging (MRI) has a limited role in the initial evaluation of the trauma patient, but may be of use for the evaluation of the spine and diaphragm in patients who are hemodynamically stable. PMID- 10798628 TI - Imaging of blunt aortic and great vessel trauma. AB - The thoracic aorta and great vessels are at risk of injury by both blunt and penetrating trauma. High-speed deceleration injury, predominately caused by motor vehicle accidents, is the primary cause of blunt traumatic aortic injury (TAI). Though largely fatal if untreated, these injuries are amenable to surgical repair if appropriately diagnosed. Algorithms for both diagnosis and treatment of TAI have undergone changes in recent years. Radiologic imaging plays a key role in the evaluation of TAI, and this review focuses on the relative roles of chest radiography, computed tomography (CT) (particularly helical CT), and aortography in the diagnostic algorithm for TAI. Other aortic imaging methods have been used in the setting of TAI, such as transesophageal echocardiography, magnetic resonance imaging, and intravascular ultrasound; although these techniques may play a complementary role in TAI evaluation, they are unlikely to have as significant an impact on routine radiologic practice as will CT. PMID- 10798629 TI - Imaging of diaphragmatic injuries. AB - Multiple imaging modalities are available for the preoperative diagnosis of diaphragmatic injury. Chest radiographs are the initial and most commonly performed imaging study to evaluate the diaphragm after trauma. When chest radiography is indeterminate, spiral computed tomography (CT) with thin sections and reformatted images is the next study of choice, particularly because most hemodynamically stable patients with blunt diaphragm injury will require an admission CT examination to evaluate the extent and anatomical sites of coexisting thoracoabdominal injuries. Magnetic resonance imaging is used to evaluate the diaphragm for patients with clinical suspicion but an indeterminate diagnosis after chest radiography and spiral CT. PMID- 10798630 TI - Chest wall trauma. AB - Sufficient trauma to the chest can result in injury to the bony thorax and soft tissues of the chest wall, increasing patient morbidity and mortality. Fractured ribs can lacerate the pleura, lung, or abdominal organs. Fractures to upper ribs, clavicle, and upper sternum can signal brachial plexus or vascular injury. Paradoxical movement of a flail chest can impair respiratory mechanics, promote atelectasis, and impair pulmonary drainage. Most patients with thoracic spine fracture-dislocations have complete neurologic deficits. Scapular fractures, associated with other injuries in almost all patients, are frequently overlooked on supine chest radiographs. Sternal fractures, associated with clinically silent myocardial contusion, are best visualized on chest computed tomography (CT). Severe trauma to the chest wall can be associated with large chest wall hematomas or collections of air within the chest wall that can communicate with the intrathoracic space. CT scanning can easily distinguish chest wall from parenchymal or mediastinal injury, whereas this differentiation my not be possible with chest radiography. PMID- 10798631 TI - Nonaortic mediastinal injuries from blunt chest trauma. AB - In addition to traumatic aortic injuries (TAI), blunt chest trauma may damage other structures in the mediastinum, including the tracheobronchial tree, the heart and pericardium, and rarely the esophagus. Tracheobronchial injuries may be difficult to separate radiographically from accompanying parenchymal lung injuries. Experience with diagnosis by computed tomography (CT) is still limited. Cardiac injuries often require emergent surgery before extensive imaging can be done. Some patients, usually those with chamber ruptures of the right heart, survive long enough to receive a chest CT, at which time hemopericardium can be detected. Upper esophageal injuries may occur in conjunction with lower cervical or upper thoracic spine injures. Distal esophageal injuries are rarely caused by blunt trauma. PMID- 10798633 TI - Computers in thoracic radiology. PMID- 10798632 TI - Imaging of penetrating thoracic trauma. AB - The management of penetrating chest injuries has evolved significantly over the past few years, with an increasing emphasis on less invasive diagnostic and therapeutic modalities. Only 15% of patients need a therapeutic operative procedure. The challenge is to detect and treat these injuries rapidly while maximizing the use of noninvasive examinations and decreasing costs. The areas potentially at risk for injury include the heart, major vessels, thoracoabdomen, neck, spine, and aerodigestive tract. A review of injuries to these areas, including the use of new diagnostic modalities such as echocardiography and computed tomography (CT) scans, are discussed. PMID- 10798634 TI - The normal CT appearances of the second carina and bronchial stump after left upper lobectomy. AB - We retrospectively evaluated the computed tomography (CT) appearance of the bronchial stump and second carina (left upper lobe spur) after left upper lobectomy. There were 69 CT examinations in 38 patients; all were free of recurrent or metastatic disease. The spur was graded as a) sharp (wedge-shaped tip configuration with <90 degrees angulation), b) lobulated (bulbous tip with <90 degrees angulation), or c) widened (>90 degrees angulation regardless of tip configuration). The bronchial stump was evaluated for the presence or absence of soft tissue in proximity to the surgical staples. The spur had a sharp appearance in 33 of 38 patients (87%) and was lobulated in 5 of 38 (13%). No patient had soft tissue at the bronchial stump. Serial examinations showed no change in the appearance of either structure. The spur remains sharp after left upper lobectomy in most patients; lobulation occurs in 13%. No changes occur over time. Interval change, widening of the spur, or soft tissue at the bronchial stump may suggest abnormality. Knowledge of normal and potentially abnormal appearances is essential to proper CT interpretation, particularly in the setting of postoperative surveillance for recurrent or metastatic disease. PMID- 10798636 TI - Radiologic findings in primary pulmonary leiomyosarcoma. AB - Leiomyosarcomas are uncommon smooth muscle tumors most commonly encountered in the female genital organs, gastrointestinal tract, and soft tissues. They are rarely encountered as primary pulmonary neoplasms. Herein we describe the conventional radiographic and computed tomographic findings in a patient with a pulmonary leiomyosarcoma. PMID- 10798635 TI - MR characteristics of progressive massive fibrosis. AB - This study was performed to evaluate the static and dynamic magnetic resonance imaging (MRI) contrast enhancement pattern of progressive massive fibrosis (PMF) in coal workers' pneumoconiosis. Eighteen lesions in 12 patients were evaluated using a 1.5-T MR unit. T1-weighted FLASH images were obtained before and 0.5, 1, 2, 3, 4, 5, 7.5, 10, 12.5, and 15 minutes after injection of gadolinium diethylenetriamine penta-acetic acid (Gd-DTPA; 0.1 mmol/kg). Imaging findings, the contrast enhancing pattern, enhancement time curve, and the contrast uptake equivalent (CE; mmol/L) were evaluated. On T1-weighted images, 14 lesions showed high signal intensity, and four showed low signal intensity. On T2-weighted images, all lesions were of low signal intensity and were indistinguishable from aerated lung parenchyma. After contrast infusion, all lesions except two enhanced markedly. The time enhancement curve showed a marked, gradual increase in signal intensity up to 3 minutes, a subtle increase in signal intensity up to 7.5 minutes, and then a plateau until 15 minutes after Gd-DTPA injection. Characteristic MR findings of PMF in coal workers' pneumoconiosis include T1 high signal intensity, T2 low signal intensity, and marked postinfusion enhancement. The time enhancement curve shows a marked, gradual increase of signal intensity up to 3 minutes and plateau up to 15 minutes. PMID- 10798637 TI - Successful transcatheter coil embolization of coronary artery fistula in an infant. AB - Congenital coronary artery fistula is a rare anomaly that can cause several types of morbidity as well as mortality. Recently, transcatheter coil embolization for congenital coronary artery fistula has been advocated as an effective alternative to surgical repair and is associated with a low morbidity and good clinical outcome. We report a 49-day-old infant who had tachycardia, tachypnea, prolonged and interrupted feeding, cardiomegaly, and continuous murmur, and who underwent successful transcatheter coil embolization for a congenital right coronary artery fistula. At review 1 year after coil occlusion showed that serial plain chest radiographs, myocardial enzyme analysis, electrocardiography, and wall motion on echocardiography were normal. PMID- 10798638 TI - Choroidal thickness changes during altered eye growth and refractive state in a primate. AB - PURPOSE: In the chick, compensation for experimentally induced defocus involves changes in the thickness of the choroid. The choroid thickens in response to imposed myopic defocus and thins in response to imposed hyperopic defocus. This study was undertaken to determine whether similar choroidal changes occur in the primate eye with induced refractive errors. METHODS: Thirty-three common marmosets were used. Eyes in 26 monkeys served as untreated control eyes, and eyes in 7 received 3 weeks of monocular lid suture to induce changes in eye growth and refractive state. Refractive errors were measured using refractometry and retinoscopy, and axial ocular dimensions, including choroidal thickness, were measured using high-frequency A-scan ultrasonography. Eyes were measured before the lids were sutured and at frequent intervals after lid opening. RESULTS: In the marmoset, choroidal thickness ranges from 88 to 150 microm and increases significantly during the first year of life. Monocular lid suture initially results in short, hyperopic eyes that then become elongated and myopic. In these animals the choroids of both the experimental and the fellow control eyes also increase in thickness with age but additionally show interocular differences that vary significantly with the relative changes in vitreous chamber depth and refraction. In eyes that are shorter and more hyperopic than control eyes the choroids are thicker, and in eyes that are longer and more myopic than control eyes the choroids are thinner. CONCLUSIONS: In marmosets, the thickness of the choroid increases during postnatal eye growth. Superimposed on this developmental increase in choroidal thickness there are changes in thickness that are correlated with the induced changes in eye size. These changes are small (<50 microm) in comparison with those observed in the chick, contributing to less than a diopter change in refractive error. PMID- 10798639 TI - Vision-dependent changes in the choroidal thickness of macaque monkeys. AB - PURPOSE: To determine whether changes in the eye's effective refractive state produce changes in the thickness of the choroid in infant monkeys. METHODS: Normal developmental changes in choroidal thickness were studied in 10 normal rhesus monkeys. Hyperopia or myopia was induced by rearing 26 infant monkeys with either spectacle or diffuser lenses secured in front of one or both eyes. The treatment lenses were worn continuously beginning at approximately 3 weeks of age for an average of 120 days. Refractive status and ocular axial dimensions, including choroidal thickness, were measured by retinoscopy and high-frequency A scan ultrasonography, respectively. RESULTS: Three lines of evidence indicate that the normal increase in choroidal thickness that occurs during early maturation can be altered by the eye's refractive state. First, in monkeys experiencing form deprivation or those in the process of compensating for imposed optical errors, choroidal thickness and refractive error were significantly correlated with eyes developing myopia having thinner choroids than those developing hyperopia. Second, the choroids in eyes recovering from binocularly induced myopia increased in thickness at a faster rate than the choroids in recovering hyperopic eyes. Third, monkeys recovering from induced anisometropias showed interocular alterations in choroidal thickness that were always in the appropriate direction to compensate for the anisometropia. These changes in choroidal thickness, which were on the order of 50 microm, occurred quickly and preceded significant changes in overall eye size. CONCLUSIONS: Changes in the eye's effective refractive state produce rapid compensating changes in choroidal thickness. Although these choroidal changes are small relative to the eye's refractive error, they may play an important role in the visual regulation of axial growth associated with emmetropization. PMID- 10798640 TI - Conjunctiva-associated lymphoid tissue in the human eye. AB - PURPOSE: Mucosa-associated lymphoid tissue (MALT) represents a part of the immune system located at mucosal surfaces. Its presence in the human eye is the point in question in the current study. Its occurrence, components, topography, and probable functional significance in the human conjunctiva and lacrimal drainage system were investigated. METHODS: Fifty-three complete conjunctival sacs were obtained from cadaveric eyes, prepared as flat wholemounts, stained, optically cleared, observed in total thickness, and sectioned for light microscopic histology, immunohistochemistry, and electron microscopy. Eight lacrimal sacs and adhering canaliculi were prepared accordingly. RESULTS: Lymphoid tissue was mainly observed in the palpebral conjunctiva, more pronounced in the upper than in the lower lid. It occurred in different forms: 1) In all specimens, diffuse lymphoid tissue of lymphocytes and plasma cells, most of which were IgA positive, formed a thin layer in the lamina propria. The overlying epithelium produced secretory component. 2) In approximately three fifths of the conjunctival sacs, organized follicular accumulations were embedded in this layer. They had a lenticular shape, were composed of B lymphocytes, and were apically covered by lymphoepithelium. 3) Both types could be associated with the conjunctival crypts. Lymphoid tissue with similar characteristics, including secondary follicles, was also observed inside the lacrimal drainage system. High endothelial venules were present in all types of lymphoid tissue. CONCLUSIONS: Human conjunctiva and lacrimal drainage system show an associated lymphoid tissue (suggesting the term conjunctiva-associated lymphoid tissue [CALT]) that contains all components necessary for a complete immune response. Expression of immunoglobulins and secretory component indicates that the conjunctiva belongs to the secretory immune system. PMID- 10798641 TI - Evidence for active control of rectus extraocular muscle pulleys. AB - PURPOSE: Connective tissue structures constrain paths of the rectus extraocular muscles (EOMs), acting as pulleys and serving as functional EOM origins. This study was conducted to investigate the relationship of orbital and global EOM layers to pulleys and kinematic implications of this anatomy. METHODS: High resolution magnetic resonance imaging (MRI) was used to define the anterior paths of rectus EOMs, as influenced by gaze direction in living subjects. Pulley tissues were examined at cadaveric dissections and surgical exposures. Human and monkey orbits were step and serially sectioned for histologic staining to distinguish EOM fiber layers in relationship to pulleys. RESULTS: MRI consistently demonstrated gaze-related shifts in the anteroposterior locations of human EOM path inflections, as well as shifts in components of the pulleys themselves. Histologic studies of human and monkey orbits confirmed gross examinations and surgical exposures to indicate that the orbital layer of each rectus EOM inserts on its corresponding pulley, rather than on the globe. Only the global layer of the EOM inserts on the sclera. This dual insertion was visualized in vivo by MRI in human horizontal rectus EOMs. CONCLUSIONS: The authors propose the active-pulley hypothesis: By dual insertions the global layer of each rectus EOM rotates the globe while the orbital layer inserts on its pulley to position it linearly and thus influence the EOM's rotational axis. Pulley locations may also be altered in convergence. This overall arrangement is parsimoniously suited to account for numerous aspects of ocular dynamics and kinematics, including Listing's law. PMID- 10798642 TI - Allelic variation in the VMD2 gene in best disease and age-related macular degeneration. AB - PURPOSE: To assess the allelic variation of the VMD2 gene in patients with Best disease and age-related macular degeneration (AMD). METHODS: Three hundred twenty one AMD patients, 192 ethnically similar control subjects, 39 unrelated probands with familial Best disease, and 57 unrelated probands with the ophthalmoscopic findings of Best disease but no family history were screened for sequence variations in the VMD2 gene by single-strand conformation polymorphism (SSCP) analysis. Amplimers showing a bandshift were reamplified and sequenced bidirectionally. In addition, the coding regions of the VMD2 gene were completely sequenced in six probands with familial Best disease who showed no SSCP shift. RESULTS: Forty different probable or possible disease-causing mutations were found in one or more Best disease or AMD patients. Twenty-nine of these variations are novel. Of the 39 probands with familial Best disease, mutations were detected in all 39 (33 by SSCP and 6 by DNA sequencing). SSCP screening of the 57 probands with a clinical diagnosis of Best disease but no family history revealed 16 with mutations. Mutations were found in 5 of 321 AMD patients (1.5%), a fraction that was not significantly greater than in control individuals (0/192, 0%). CONCLUSIONS: Patients with the clinical diagnosis of Best disease are significantly more likely to have a mutation in the VMD2 gene if they also have a positive family history. These findings suggest that a small fraction of patients with the clinical diagnosis of AMD may actually have a late-onset variant of Best disease, whereas at the same time, a considerable fraction of isolated patients with the ophthalmoscopic features of Best disease are probably affected with some other macular disease. PMID- 10798643 TI - Altered expression of secreted frizzled-related protein-2 in retinitis pigmentosa retinas. AB - PURPOSE: Inherited retinal degenerations such as retinitis pigmentosa (RP) are characterized by progressive death of the photoreceptors due to apoptosis. To identify changes in gene expression associated with the degenerative state in RP retinas, expression profiling of apoptosis-related genes was performed using a gridded array technique. METHODS: Total RNAs from RP and control retinas were used to generate radiolabeled cDNA probes to screen gridded membrane arrays of 205 apoptosis-related genes. Reverse transcription-polymerase chain reaction was used to generate probes corresponding to differentially expressed genes for Northern blot analysis and for mRNA in situ hybridization studies of retinal cryosections. Fluorescence immunocytochemistry was performed on retinal sections using available antibodies. RESULTS: By expression profiling, we identified upregulated expression of the mRNA for secreted Frizzled-related protein-2 (SFRP2) in RP retina in comparison with control. By Northern blot analysis, SFRP2 mRNA levels were 2- to 20-fold higher in RP samples than in controls. The localization of SFRP2 mRNA by in situ hybridization varied according to the degree of degeneration, from stratified in relatively well-preserved retinas to diffuse in the highly degenerative state. By immunofluorescence, SFRP2 protein in RP retinas was found mainly to colocalize with the cell adhesion and signal transducing protein beta-catenin. CONCLUSIONS: SFRPs can regulate apoptosis in vitro and appear to interact with the Wnt/Frizzled signaling pathway, which includes routes to apoptotic activation. Increased SFRP2 expression in RP retinas suggests that an altered pattern of Wnt signal transduction may be a step in the degenerative process linking causal mutations with eventual photoreceptor demise. PMID- 10798644 TI - BIGH3 exon 14 mutations lead to intermediate type I/IIIA of lattice corneal dystrophies. AB - PURPOSE: To screen the BIGH3 gene in three unrelated families with lattice corneal dystrophy (LCD), two of which disclosed a particular phenotype. METHODS: Genomic DNA was extracted from peripheral leukocytes of the affected patients and their family members. The entire coding sequence of the BIGH3 gene was screened for mutations by means of transcript analysis on total RNA isolated from peripheral leukocytes by reverse transcription-polymerase chain reaction performed with primers designed for this study. Each mutation was confirmed at the genomic level, by using published primers. RESULTS: One family that had a typical form of LCD, had the described R124C mutation in the BIGH3 gene. Two families with atypical forms of LCD were negative for the previously known mutations of the gene. Direct sequencing of the BIGH3 mRNA in the latter two families allowed us to identify two mutations located in exon 14. They consist of a 9-bp insertion at position 18851886 and one missense mutation at position 1877 of the BIGH3 gene. Three new polymorphisms were also observed. CONCLUSIONS: Two mutations different from those linked to LCD have been found in clinically distinguishable forms of this disease, intermediate between LCDs types I and IIIA. The DNA segment comprising both alterations normally encodes for a highly conserved region of the fourth internal domain of the Betaig-h3 protein, suggesting that this region may be of functional and/or structural importance. The identification of new mutations by screening of the complete BIGH3 gene and the comparative analysis of the induced modifications in betaig-h3 protein should shed light in the understanding of the molecular mechanisms underlying LCDs resulting from mutations in the BIGH3 gene, and may help to explain their phenotypic heterogeneity. PMID- 10798645 TI - Visual function and subjective quality of life compared in subjects with acquired macular disease. AB - PURPOSE: To determine the objective measures of visual function that are most relevant to subjective quality of vision and perceived reading ability in patients with acquired macular disease. METHODS: Twenty-eight patients with macular disease underwent a comprehensive assessment of visual function. The patients also completed a vision-related quality-of-life questionnaire that included a section of general questions about perceived visual performance and a section with specific questions on reading. RESULTS: Results of all tests of vision correlated highly with reported vision-related quality-of-life impairment. Low-contrast tests explained most of the variance in self-reported problems with reading. Text-reading speed correlated highly with overall concern about vision. CONCLUSIONS: Reading performance is strongly associated with vision-related quality of life. High-contrast distance acuity is not the only relevant measure of visual function in relation to the perceived visual performance of a patient with macular disease. The results suggest the importance of print contrast, even over print size, in reading performance in patients with acquired macular disease. PMID- 10798646 TI - Detection of sialomucin complex (MUC4) in human ocular surface epithelium and tear fluid. AB - PURPOSE: To evaluate human ocular surface epithelium and tear fluid for the presence of sialomucin complex (MUC4), a high-molecular-weight heterodimeric glycoprotein composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis assays were used to identify sialomucin complex RNA in ocular surface epithelia. Immunoprecipitation and immunoblot analysis were used to identify immunoreactive species in human tears and in the corneal and conjunctival epithelia using antibodies specific for carbohydrate and peptide epitopes on the sialomucin complex subunits. Immunofluorescence staining was used to detect sialomucin complex in frozen sections and impression cytology specimens of human cornea and conjunctival epithelia. RESULTS: ASGP-1- and ASGP-2-specific sequences were amplified from RNA extracted from both conjunctival and corneal epithelial biopsies by RT-PCR. Sialomucin complex transcripts were also detected in these tissues by Northern blot analysis, with a greater level of RNA detected in the peripheral than the central corneal epithelium. Sialomucin complex was immunoprecipitated from tear fluid samples and both corneal and conjunctival epithelia and detected by immunoblot analysis with specific anti-ASGP-1 and anti ASGP-2 antibodies. The ASGP-1 peptide antibody HA-1 stained the full thickness of the corneal and conjunctival epithelia. In contrast, antibody 15H10, which reacts against a carbohydrate epitope on ASGP-1, stained only the superficial epithelial layers of these tissues. No staining was observed in the conjunctival goblet cells. CONCLUSIONS: Sialomucin complex was originally identified in rat mammary adenocarcinoma cells and has recently been shown to be produced by the ocular surface epithelia of rats. Furthermore, it has been identified as the rat homologue of human MUC4 mucin. The present studies show that it is expressed in the stratified epithelium covering the surface of the human eye and is present in human tear fluid. Expression of a carbohydrate-dependent epitope on the mucin subunit (ASGP-1) of sialomucin complex occurs in a differentiation-dependent fashion. Sialomucin complex joins MUC1 as another membrane mucin produced by the human ocular surface epithelia but is also found in the tear fluid, presumably in a soluble form, as found on the rat ocular surface. PMID- 10798647 TI - Modulation of TNF-alpha-induced apoptosis in corneal fibroblasts by transcription factor NF-kappaB. AB - PURPOSE: Previous studies have suggested no role for tumor necrosis factor (TNF) alpha in the modulation of apoptosis in corneal fibroblasts. However, recent investigations have demonstrated that nuclear factor (NF)-kappaB activation by TNF-alpha mediates negative apoptotic effects that must be blocked to unmask the apoptotic effects of TNF-alpha in vitro. The purpose of this study was to investigate the role of transcription factor NF-kappaB in the suppression of TNF alpha-induced apoptosis of corneal fibroblasts. METHODS: mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay. Proteins were detected by immunocytochemistry and immunoprecipitation with Western blot analysis. Cell death was evaluated by trypan blue exclusion assay in corneal fibroblasts treated with TNF-alpha in presence or absence of the specific inhibitor of NF-kappaB activation, SN50, actinomycin D, or actinomycin D with dexamethasone, ketorolac tromethamine, or diclofenac sodium. Apoptosis was monitored by trypan blue exclusion, colorimetric cell assay, CPP32 activation assay, DNA fragmentation assay, and transmission electron microscopy. NF-KB activation was monitored using electrophoretic gel shift assay. RESULTS: TNF alpha, TNF receptor (R)I, and TNFRII mRNAs were detected in all three cultured corneal cell types and in ex vivo corneal epithelium using RT-PCR. TNF-alpha mRNA was also detected in ex vivo corneal epithelium, corneal epithelial cells, and stromal fibroblasts with the RNase protection assay. TNF-alpha, TNFRI, and TNFRII proteins were detected by immunocytochemistry in all three major corneal cell types in human corneal tissue. TNF-alpha protein was also detected in ex vivo corneal epithelium, primary corneal epithelial cells, and primary stromal fibroblasts using immunoprecipitation and Western blot analysis. TNF-alpha stimulated corneal fibroblast cell death when NF-kappaB activation was blocked with actinomycin D or SN50. Enhanced cell death was noted with dexamethasone, ketorolac tromethamine, or diclofenac sodium when used in the presence, but not in the absence, of actinomycin D. A gel shift assay revealed induction of NF-KB by TNF-alpha and suppression of induction in the presence of actinomycin D or SN50, but not by the control peptide SN50M. CONCLUSIONS: The TNF-alpha receptor system is expressed in the cornea, and NF-kappaB activation is an important regulator of TNF-alpha-mediated corneal fibroblast apoptosis. Nonsteroidal anti inflammatory agents or corticosteroids may potentiate corneal fibroblast apoptosis in response to cytokine stimulation. PMID- 10798648 TI - Role of cell adhesion-associated protein, pinin (DRS/memA), in corneal epithelial migration. AB - PURPOSE: To determine whether the cellular distribution of cell adhesion associated protein, pinin, is altered during corneal epithelial migration in response to debridement wounding and to determine the effect of overexpression of pinin in cultured epithelial cells. METHODS: Corneas from guinea pig and embryonic (day 17) chickens were excised, wounded, and placed on organ-culture rafts. At time points from 0 to 24 hours, corneas were cryosectioned and subsequently analyzed by immunofluorescence or immunoelectron microscopy for the presence and distribution of pinin. Cultured epithelial cell line MDCK (Madin Darby canine kidney) confluent monolayers were wounded by scraping and examined by immunofluorescence for pinin and desmoplakin. MDCK cells were transfected with full-length pinin cDNA. After selection in Geneticin, clones of pinin-transfected cells were isolated. Monolayers of transfected cells were scrape-wounded and assayed for their ability to migrate. RESULTS: Within 2 hours after wounding, although morphologically identifiable desmosomes were present on migrating epithelial cells, the association of pinin to desmosomes was greatly reduced. Finally, after completion of wound closure, pinin returned to the corneal epithelial desmosome. Wounding of confluent epithelial monolayers (MDCK) in vitro demonstrated a very similar change in the distribution of pinin, whereas desmoplakin remained cell boundary-associated. Transfection of pinin into cultured epithelial cells resulted in an overexpression of pinin. Clones of cells expressing high levels of pinin exhibited marked reduction in their ability to migrate after wounding. CONCLUSIONS: Pinin is involved in corneal epithelium migration. The localization of pinin at or near the desmosome is correlated with the epithelial quiescence. The loss of pinin from the cell boundary correlates with the transition from quiescence to actively migrating. Overexpressing pinin in cultured epithelial cells affects epithelial homeostasis and, in turn, drives the epithelial cells to a hyperstable epithelial adhesive state and inhibits the transition from quiescence to migratory. PMID- 10798649 TI - Activation of epidermal growth factor receptor during corneal epithelial migration. AB - PURPOSE: Epidermal growth factor (EGF) and related growth factors: transforming growth factor (TGF)-alpha, heprin-ding (HB)-EGF, and amphiregulin (AR), have been shown to stimulate events associated with epithelial wound repair. These growth factors function by binding to a common EGF receptor (EGFR), tyrosine kinase. We have used in vivo and organ culture wound-healing models to examine the kinetics and extent of EGFR activation during corneal epithelial wound repair and whether the epithelium itself produces EGFR ligands capable of stimulating the healing process. METHODS: In the in vivo model, 3-mm debridement wounds were made in rat corneas and allowed to heal in situ. Activation of EGFR was analyzed by 1) indirect immunofluorescence microscopy, 2) immunoprecipitation using anti-EGFR and anti-phosphotyrosine (anti-PT), and 3) binding-site localization using EGF fluorescein isothiocyanate (FITC). Relative levels of mRNA for EGF, TGF-alpha, HB EGF, and AR were determined using reverse transcription-polymerase chain reaction. To determine whether inhibiting EGFR activation slows epithelial migration, wounded corneas were allowed to heal in organ culture in the presence of tyrphostin AG1478 (0-50 microM), a specific inhibitor of EGFR kinase activity. RESULTS: In unwounded corneas, EGFR was localized in basal cells and appeared to be membranous. Within 1 hour after wounding, EGFR was no longer immunolocalized in the membranes of cells migrating into the wound area. EGF-FITC-binding assays indicated that EGFR ligands could penetrate all the way to the limbus. Immunoprecipitation showed that EGFR was phosphorylated on tyrosine residues within 30 minutes after wounding and that phosphorylation levels increased after wounding. Levels of mRNA for TGF-alpha, HBEGF, and AR all appeared to increase after wounding. In organ culture experiments, tyrphostin AG1478 inhibited migration rates in a dose-dependent manner. CONCLUSIONS: These data indicate that EGFR was activated during corneal epithelial wound healing in vivo. Furthermore, this activation appears to be a necessary component of the process, because inhibition of the EGFR signaling cascade significantly slowed migration rates. PMID- 10798650 TI - Flow cytometric analysis of inflammatory markers in conjunctival epithelial cells of patients with dry eyes. AB - PURPOSE: To investigate in impression cytology (IC) specimens the expression of inflammatory and apoptosis-related markers by conjunctival epithelial cells from patients with dry eye as a rationale for treatment with topical cyclosporine. METHODS: Immunologic anomalies were identified at baseline, before treatment with the masked medication, in a homogeneous series of patients with dry eye syndrome, who were enrolled in a large European multicenter clinical trial (Cyclosporin A Dry Eye Study; Allergan, Irvine, CA). IC specimens were collected in 243 patients with moderate to severe keratoconjunctivitis sicca (KCS), with or without Sjogren's syndrome (SS). Fifty normal subjects were separately examined to provide normal control values. Specimens were analyzed in a masked manner by flow cytometry, using antibodies directed to markers of the immune system and/or apoptotic pathway: HLA DR, CD40, CD40 ligand, Fas, and APO2.7. Levels of expression were quantified, and results were compared with those obtained in the 50 normal patients. RESULTS: One hundred sixty-nine specimens were successfully interpreted at baseline, including 41% from patients with SS. A highly significant increase of HLA DR expression by conjunctival cells was found in KCS affected eyes compared with normal eyes, which did not express this marker or did so very weakly. HLA DR expression in eyes with SS was significantly higher than in KCS-affected eyes without SS. Fas and APO2.7 were found at low levels in all normal and KCS-affected eyes. CD40 and CD40 ligand expressions were significantly increased in eyes with KCS compared with normal eyes. HLA DR, CD40 and Fas were found at significantly higher levels in the SS group than in the non-SS group. CONCLUSIONS. Conjunctival cells from patients with dry eye with moderate to severe KCS, with or without SS, overexpress inflammatory and apoptosis-related markers. Whether inflammation is a primary phenomenon in KCS or is the consequence of repetitive abrasion of the ocular surface after tear film deficiency remains to be determined. These data, nevertheless, support the use of immunomodulatory and/or anti-inflammatory drugs in the treatment of patients with KCS. PMID- 10798651 TI - Telomerase activity and p53 expression in pterygia. AB - PURPOSE: To investigate tolomerase activity and p53 expression in pterygial tissue. METHODS: Pterygia tissue was obtained during excisional surgery fr om 35 eyes of 35 patients, and superior bulbar conjunctival tissue from the same eye was also sampled as control when possible. Fluorescence telomeric repeat amplification protocol was used to measure telomerase activity in whole pterygium samples from 9 cases and in the epithelium and stroma of pterygium from another 10 cases. p53 protein content was measured by enzyme-linked immunosorbent assay (ELISA) in tissues obtained from 7 eyes, as well as in epithelial cell suspensions collected by brush cytology in 8 eyes. Six samples were also analyzed for UV-specific mutations in the p53 gene by the single-strand conformation polymorphism technique and DNA sequencing. A conjunctival epithelial cell line was irradiated with sublethal levels of UV-B to investigate whether telomerase activity can be induced in vitro. RESULTS: In all, 63% of pterygia samples demonstrated telomerase activity, whereas all 10 paired conjunctival control samples were negative (P = 0.05, chi-square test). Of the 10 samples in which telomerase activity was measured separately in the epithelium and stroma of pterygia, 5 samples were positive in the epithelium, only 1 of which had activity in the stroma. Average telomerase activity in positive samples was 18.44 +/- 8.77 U/microg protein, compared with telomerase activity measured in a carcinoma in situ patient (33.73 U/microg), and in an immortalized conjunctival epithelial cell line (50.72 +/- 15.55 U/microg). Telomerase activity was not upregulated in this cell line by UV-B exposure. All 6 pterygia samples tested for p53 mutations did not reveal the UV-specific mutations in exons 5, 6, 7, or 8. No statistical significance was observed in the pterygium or conjunctiva p53 protein levels in epithelial cells collected by brush cytology, while p53 protein level was lower in pterygia when measured in whole tissue samples. CONCLUSIONS: Telomerase activity was detected in some pterygia, mostly in the epithelium. Pterygia was not associated with an increase in epithelial p53 protein content measured by ELISA. PMID- 10798652 TI - Experimental glaucoma and cell size, density, and number in the primate lateral geniculate nucleus. AB - PURPOSE: To examine the effects that elevated intraocular pressure (IOP), a glaucoma risk factor, has on the size, density, and number of neurons in the primate lateral geniculate nucleus (LGN). METHODS: The monkey model of experimental glaucoma was combined with standard histologic staining and analysis techniques. Fourteen animals were examined. RESULTS: Mean IOPs higher than 40 mm Hg for 2.5, 4, 8, and 24 weeks resulted in reductions of 10% to 58% in the cross sectional areas of LGN neurons receiving input from the glaucomatous eye. Reductions for animals with lower mean IOPs (37 and 28 mm Hg) for 16 and 27 weeks were 16% and 30%, respectively. Neurons receiving input from the normal eye also were reduced in size (4 -26%). No differential effect in cell size was seen for magnocellular versus parvocellular neurons. Elevation of IOP resulted in an increase in cell density in all layers of the LGN. The increase was approximately two times greater in parvocellular (59%) than magnocellular (31%) layers. When corrected for volumetric shrinkage of the LGN, the estimated loss of neurons was approximately four times greater in the magnocellular than parvocellular layers (38% versus 10%). CONCLUSIONS: Elevation of IOP affects the size, density, and number of neurons in the LGN, and the volume of the nucleus itself. Although higher mean pressures (more than 40 mm Hg) reduce the period during which these changes occur, comparable damage can be achieved by even moderate (28 -37 mm Hg) levels of elevated IOP. On the basis of cell loss, elevation of IOP appears to have a more profound degenerative effect on the magnocellular than on the parvocellular regions of the LGN. PMID- 10798653 TI - Patterns of intraocular pressure elevation after aqueous humor outflow obstruction in rats. AB - PURPOSE: To determine the diural intraocular pressure (IOP) response of Brown Norway rat eyes after sclerosis of the aqueous humor outflow pathways and its relationship to optic nerve damage. METHODS: Hypertonic saline was injected into a single episcleral vein in 17 animals and awake IOP measured in both the light and dark phases of the circadian cycle for 34 days. Mean IOP for light and dark phases during the experimental period were compared with the respective pressures of the uninjected fellow eyes. Optic nerve cross sections from each nerve were graded for injury by five independent masked observers. RESULTS: For fellow eyes, mean light- and dark-phase IOP was 21 +/- 1 and 31 +/- 1 mm Hg, respectively. For four experimental eyes, mean IOPs for both phases were not altered. Six eyes demonstrated significant mean IOP elevations only during the dark phase. Of these, five showed persistent, large circadian oscillations, and four had partial optic nerve lesions. The remaining seven eyes experienced significant IOP elevations during both phases, and all had extensive optic nerve damage. CONCLUSIONS: Episcleral vein injection of hypertonic saline is more likely to increase IOP during the dark phase than the light. This is consistent with aqueous outflow obstruction superimposed on a circadian rhythm of aqueous humor production. Because these periodic IOP elevations produced optic nerve lesions, both light- and dark-phase IOP determinations are necessary for accurate correlation of IOP history to optic nerve damage in animals housed in a light- dark environment. PMID- 10798654 TI - Truncations in the TIGR gene in individuals with and without primary open-angle glaucoma. AB - PURPOSE: To investigate the coding exons in the trabecular meshwork-induced glucocorticoid response protein (TIGR) gene for mutations in primary open-angle glaucoma (POAG) in Chinese subjects. METHODS: Ninety-one Chinese patients with POAG and 113 of their family members without glaucoma were screened for sequence alterations in the TIGR gene by polymerase chain reaction, conformation-sensitive gel electrophoresis, and DNA sequencing. One hundred thirty-two unrelated individuals without glaucoma, aged 50 years or more, were studied as control subjects. RESULTS: Five sequence variants that lead to amino acid changes were identified. One was novel: Arg91Stop in one patient with POAG. Four had been reported: Arg46Stop in subjects with and without POAG, including an unaffected 77 year-old woman homozygous for Arg46Stop; Gly12Arg in subjects without glaucoma; and Asp208Glu and Thr353Ile in subjects with and without POAG. The previously reported 1-83(G-->A) and Arg76Lys polymorphisms were detected in both patients and controls and always occurred together. CONCLUSIONS: A different pattern of TIGR sequence variants exists in the Chinese than in non-Chinese populations. No common TIGR mutation that causes POAG was found. The occurrence of subjects without glaucoma who are heterozygous or homozygous for Arg46Stop suggests that reduction in the amount of TIGR protein does not cause glaucoma. Thus, the TIGR missense mutations known to cause POAG probably do not cause glaucoma by inactivating a normal TIGR function, but rather through the gain of a pathologic function. PMID- 10798655 TI - Herpes simplex virus-mediated gene delivery to the rodent visual system. AB - PURPOSE: To determine the types of cells in the visual system of the mouse and rat that can express a transgene delivered by an attenuated replication competent Herpes simplex virus-1 (HSV-1) vector. METHODS: C57/BL6 x BALB/C mice and Albino rats were treated with 1 x 10(7) pfu of the HSV-1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. The hrR3 virus was delivered by topical application to the cornea, intravitreal (IV) injection, intracameral injection (IC), or stereotactic injection into the visual cortex (VC). At specified times postinfection, animals were killed and tissues were removed, fixed, sectioned, and stained with X-gal or hematoxylin and eosin for histochemical and histopathologic examination. RESULTS: Topical delivery after corneal scarification in both mouse and rat resulted in lacZ expression in 25% of the corneal epithelial cells and 25% of the retinal pigment epithelium (RPE) cells. Topical application without concurrent scarification also resulted in transgene delivery to 20% of the RPE cells of the rat but not the mouse. IV injection resulted in expression primarily in RPE cells, with up to 100% of the cells expressing lacZ in the mouse and rat. Other cells expressing the transgene included ciliary body (CB) and optic nerve cells. Up to 25% of the retinal ganglion cells in the rat expressed lacZ, but only rarely in mice. IC delivery in rats resulted in expression in trabecular meshwork, CB cells, RPE, and iris epithelium. Injection into area 17 of the rat VC resulted in efficient labeling of the VC neurons and neurons in the lateral geniculate nucleus without any evident pathology or inflammation. Neither inflammation nor disease pathology was observed in either the mouse or rat after any route of delivery. CONCLUSIONS: It was demonstrated that the hrR3 HSV-1 virus can deliver a functioning gene to several cell types in the eye and neurons in the VC and that the virus can move via retrograde transport to nuclei that project to the VC. PMID- 10798656 TI - Role of macrophages in restricting herpes simplex virus type 1 growth after ocular infection. AB - PURPOSE: To investigate the role macrophages play in controlling herpes simplex virus (HSV)-1 replication after infection of the murine cornea. METHODS: Macrophage depletion in selected tissues before or after virus infection was achieved by repeated subconjunctival (SCJ) and/or intravenous (IV) injection of liposomes containing dichloromethylene diphosphonate (L-Cl2MDP). Controls received liposomes containing phosphate-buffered saline (L-PBS). The efficiency of depletion was evaluated by histologic examination. Virus content in infected tissues was determined by standard plaque assay. Delayed-type hypersensitivity (DTH) responsiveness was assessed using the ear-swelling assay. Antibody isotype responses to virus antigens and cytokine production were monitored by enzyme linked immunosorbent assay. RESULTS: Balb/c mice given SCJ injection of L-Cl2MDP 4 and 2 days before HSV-1 corneal infection were found to have ocular virus titers as much as 10(5)-fold higher than that seen in the L-PBS-treated controls 8 days after infection. When L-Cl2MDP treatment was delayed until 2 and 4 days after infection, virus titers in the eye were analogous to those in the control animals. Subconjunctival and submandibular lymph node macrophages in mice given local (SCJ) L-Cl2MDP pretreatment were profoundly reduced, whereas the number of corneal Langerhans' cells and lymph node dendritic cells remained unchanged. Local L-Cl2MDP pretreatment was associated with significantly reduced DTH responsiveness to HSV-1 antigen, and an alteration in selected antibody isotype production. Depletion of macrophages in the subconjunctival tissue before corneal infection was not accompanied by enhanced virus growth at early times (2 or 4 days) after infection. CONCLUSIONS: Macrophages play an important role in restricting HSV-1 growth after corneal infection. These cells appear to be required for the development of an acquired immune response, presumably by functioning in antigen processing and presentation. The hypothesis that macrophages are major participants in innate immunity to HSV-1 corneal infection was not supported. PMID- 10798657 TI - Analysis of in vivo regulatory properties of T cells activated in vitro by TGFbeta2-treated antigen presenting cells. AB - PURPOSE: To determine whether naive T cells activated in vitro by antigen-pulsed, transforming growth factor(beta) (TGFbeta)-treated antigen presenting cells (APCs) acquire the capacity to suppress the induction and expression of delayed hypersensitivity in vivo. METHODS: Naive ovalbumin (OVA)specific T cells from DO 11.10 Tcr transgenic mice were stimulated in vitro with OVA-pulsed TGF(beta2) treated APCs. The cultured cells were harvested and assayed for in vitro production of mature TGFbeta. Similar cells were coinjected with primed OVA specific BALB/c T cells plus OVA-pulsed APCs into ear pinnae of normal BALB/c mice (assay for delayed hypersensitivity expression) or coinjected with OVA pulsed APCs into footpads of naive DO11.10 mice whose draining lymph node cells were harvested 4 days later and assayed in vitro for capacity to secrete interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) when stimulated with OVA (assay for induction of delayed hypersensitivity). RESULTS: DO11.10 T cells activated in vitro by OVA-pulsed TGFbeta2-treated APCs secreted large amounts of mature TGFbeta and suppressed the expression of delayed hypersensitivity in a local adoptive transfer assay. Suppression was reversed in the presence of neutralizing anti-TGFbeta antibodies. In addition, in vitro generated regulatory T cells influenced naive T cells in DO11.10 mice that were responding to an initial immunization with OVA to secrete IL-4, rather than IFN-gamma. This influence was independent of TGFbeta. CONCLUSIONS: OVA-pulsed APCs, pretreated in vitro with TGF(beta)2, activate DO11.10 T cells in a manner that endows the responding cells with the capacity to suppress the induction and then the expression of delayed hypersensitivity in vivo. In certain ways, these properties of in vitro-activated DO11.10 T cells resemble the properties of afferent and efferent regulatory T cells typically found in the spleens of animals with anterior chamber-associated immune deviation. PMID- 10798659 TI - Lysostaphin treatment of methicillin-resistant Staphylococcus aureus keratitis in the rabbit. AB - PURPOSE: To determine the efficacy of lysostaphin treatment of methicillin sensitive and methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a rabbit model. METHODS: The sensitivity to lysostaphin and vancomycin were compared for 34 MRSA and 12 methicillin-sensitive strains. Methicillin-resistant S. aureus strain 301 (MRSA 301) or a methicillin-sensitive strain of low virulence, ISP546, was intrastromally injected into rabbit corneas. Rabbit eyes were treated topically every 30 minutes from 4 to 9 or 10 to 15 hours postinfection with 0.28% lysostaphin or 5.0% vancomycin. Rabbits were killed and corneas were excised and cultured to determine the number of colony forming units (CFU) per cornea. RESULTS: Ninety percent minimal inhibitory concentrations were at least 19-fold lower for lysostaphin than for vancomycin. With early therapy (4 -9 hours postinfection) lysostaphin sterilized all MRSA 301-infected corneas, whereas untreated corneas contained 6.52 log CFU/cornea (P < or = 0.0001). Corneas infected with MRSA 301 and treated similarly with vancomycin retained 2.3 +/-0.85 log CFU/cornea, and none were sterile. When therapy was begun later (10 15 hours postinfection) the residual bacteria in lysostaphin-treated eyes were significantly less numerous than in vancomycin-treated eyes (0.58 +/- 0.34 vs. 5.83 +/- 0.16 log CFU/cornea, respectively; P < or = 0.0001). Three experiments were performed to demonstrate that lysostaphin penetrated the cornea to kill bacteria in vivo; lysostaphin-treated eyes were found to recover from infection, bacteria that did not cause epithelial defects (ISP546) were susceptible to lysostaphin, and inhibition of lysostaphin when harvesting corneas did not alter the observed therapeutic values of lysostaphin. CONCLUSIONS: Lysostaphin is very effective in treating keratitis mediated by methicillin-sensitive or methicillin resistant S. aureus. PMID- 10798658 TI - Langerhans cells, orthotopic corneal allografts, and direct and indirect pathways of T-cell allorecognition. AB - PURPOSE: To determine after orthotopic corneal allografting the role of Langerhans cells in activation of T cells via the direct and indirect pathways of allorecognition and the relationship between these pathways and the rapidity of graft rejection. METHODS: Corneas from eyes of normal mice and from eyes after superficial cauterization were grafted to eyes of major histocompatibility complex (MHC) and/or minor histocompatibility (H)-disparate recipient mice. The grafts were analyzed through time for content of class 1 MHC- bearing Langerhans cells and for rejection or acceptance. Graft recipients were evaluated for acquisition of delayed hypersensitivity (DH) and cytotoxic T cells (Tc) directed at donor MHC and minor H alloantigens. RESULTS: Langerhans cells migrated more rapidly into epithelium of cauterized grafts than normal grafts. Unlike normal grafts, the vast majority of cauterized allografts were rejected within 2 weeks. Normal grafts induced neither DH nor Tc directed at donor MHC antigens, whereas cauterized grafts induced both DH and Tc specific for donor MHC. All grafts induced DH directed at donor minor H antigens, but only rejected grafts correlated with acquisition of Tc directed at donor minor H antigens. CONCLUSIONS. The rapidity of orthotopic corneal allograft rejection correlated with density of Langerhans cells within epithelium and with acquisition of donor specific DH and Tc. Although recipient-derived Langerhans cells promoted minor H specific, self-MHC-restricted T cells (indirect pathway) and subacute graft rejection, donor-derived Langerhans cells promoted early, acute rejection in conjunction with allogeneic MHC-specific Tc (direct pathway). PMID- 10798660 TI - PCR-RFLP-mediated detection and speciation of bacterial species causing endophthalmitis. AB - PURPOSE: To determine the usefulness of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in the identification and speciation of bacteria causing endophthalmitis. METHODS: PCR-RFLP was performed on 53 strains of 14 bacterial species (eight Gram positive and five Gram negative) collected from both keratitis and endophthalmitis patients. Two pairs of oligonucleotide primers based on the 16S rDNA gene were used to PCR-amplify 1.2- and 1.0-kb fragments of bacterial genomic DNA. RFLPs within the PCR product were used to speciate the organisms. RESULTS: The sensitivity of the nested PCR amplification reaction was one organism. All bacteria tested could be identified and speciated using RFLP analysis except for Escherichia coli and Serratia marcescens, which could not be interdifferentiated using RFLP. Molecular analysis of two vitreous samples from two eyes with typical signs of bacterial endophthalmitis confirmed the presence of E. coli in the vitreous from a culture-positive case with E. coli endophthalmitis and revealed the presence of Staphylococcus epidermidis in the vitreous of a culture-negative case. CONCLUSIONS: It is expected that this technique will provide a useful laboratory tool for future microbiologic diagnosis of patients presenting with endophthalmitis, especially for those eyes that prove culture negative. PMID- 10798661 TI - Synergistic induction of eotaxin expression in human keratocytes by TNF-alpha and IL-4 or IL-13. AB - PURPOSE: To investigate the effects of tumor necrosis factor (TNF)-alpha, interleukin (IL)4, and IL-13 on expression of the chemokine eotaxin by cultured human keratocytes. METHODS: Cultured human keratocytes were incubated with various combinations and concentrations of TNF-alpha, IL-4, and IL-13. The concentration of eotaxin in the culture supernatant was subsequently measured by enzyme-linked immunosorbent assay, and the amount of eotaxin mRNA in cell lysates was determined by reverse transcription-polymerase chain reaction analysis. RESULTS: Keratocytes incubated in the absence of cytokines did not release detectable amounts of eotaxin into the culture medium. Whereas incubation of keratocytes with TNF-alpha, IL-4, or IL-13 alone or with the combination of IL-4 and IL-13 had only a small effect on eotaxin release, exposure of the cells to TNF-alpha in combination with either IL-4 or IL-13 resulted in a marked increase in eotaxin production that was both time and dose dependent. The abundance of eotaxin mRNA in keratocytes was also increased in a synergistic manner by incubation of cells with TNF-alpha together with either IL-4 or IL-13. CONCLUSIONS: Stimulation of human keratocytes with the combination of TNF-alpha and either IL-4 or IL-13 resulted in synergistic increases in both the abundance of eotaxin mRNA and the release of eotaxin protein. This cytokine-induced increase in eotaxin production by keratocytes may contribute to eosinophil infiltration in inflammatory ocular diseases such as vernal keratoconjunctivitis. PMID- 10798662 TI - Age-related changes in the absorption characteristics of the primate lens. AB - PURPOSE: To quantitate aging of the primate lens by changes in the absorption characteristics that are related to the yellowing of lens protein. METHODS: The lenses of lower primates and humans were sectioned anterior to posterior every 0.25 mm, and the UV-visible spectrum of each section was measured to determine the cumulative spectra along the visual axis. The ratio of the absorbance at 320 nm (formed with aging) to the absorbance at 365 nm (present in the young lens) was correlated with the age of the lens. RESULTS: In the young primate UV-B is transmitted to the retina, and UV-A is transmitted to the nucleus of the lens. By puberty, changes in the absorption characteristics of the lens that are associated with the yellowing of lens protein prevented most of the UV-B from reaching the retina and by the eighth decade, the transmittances at 320 and 365 nm to the nucleus of the lens were approximately 40% and 79%, respectively. A linear relationship between the ratio of absorbance at 320 to 365 nm and age was found for both lower primates and humans to the age of 80 years. This is surprising, because the maximum life span of the lower primate is approximately 35 years, whereas humans may live 100 years. CONCLUSIONS: These data suggest that the observed spectral changes associated with the yellowing of the lens are the result of a chronological process, such as chemical or photochemical modifications, not biological aging. PMID- 10798664 TI - Overexpression of Na(+)-dependent myo-inositol transporter gene in mouse lens led to congenital cataract. AB - PURPOSE: Maintaining appropriate osmotic pressure is essential for maintaining lens transparency. This study was performed to investigate whether high levels of myo-inositol, one of the major organic osmolytes in the lens, would lead to cataract development. METHODS: Transgenic mouse lines carrying the bovine Na(+) dependent myo-inositol transporter (bSMIT) cDNA under the control of the mouse alphaA-crystallin promoter were generated. RESULTS. Increased bSMIT expression was accompanied by increased myo-inositol level in the lens and increased uptake of (3H) myo-inositol by the lens in culture. The transgenic mice developed observable cataract under normal rearing conditions beginning at 2 to 8 weeks of age, and the severity of cataract development was correlated to the level of bSMIT gene expression and lens myo-inositol accumulation. For transgenic mouse line 3352, heterozygous mice did not develop cataract, whereas homozygous ones did. Prenatal feeding of heterozygous 3352 mice with high myo-inositol diet led to cataract development, indicating that cataract development was not merely due to a nonspecific effect of SMIT overexpression. Introducing aldose reductase overexpressing transgene into heterozygous 3352 mice also led to cataract development, indicating that this type of cataract is primarily due to osmotic stress. CONCLUSIONS: The present results indicate that high levels of myo inositol and sorbitol in the lens contribute to cataract development. This is a useful model to study the role of osmotic stress in cataractogenesis during lens development. PMID- 10798663 TI - Contribution of calpain Lp82-induced proteolysis to experimental cataractogenesis in mice. AB - PURPOSE: The purpose of the present experiments was to provide a biochemical mechanism for the involvement of lens-specific calpain Lp82 in experimental cataractogenesis in mice. METHODS: Nuclear cataracts were produced by culturing lenses from 4-week-old mice and rats in calcium ionophore A23187 or by injection of buthionine sulfoximine (BSO) into 7-day-old mice. Casein zymography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, calcium determinations, in vitro precipitation, and cleavage site analysis by mass spectrometry were performed on lens samples. RESULTS: Amino acid sequences for Lp82 were found to be highly conserved in lenses from mouse to cow, and expressed Lp82 proteolytic activity was high in the mouse and rat. Lenses from mice were more susceptible to A23187-induced cataract and BSO cataracts than rats. Both types of cataracts showed rapid elevation of calcium, activation of Lp82 and m calpain, and proteolysis of crystallins. Lp82 caused in vitro precipitation of crystallins; and in contrast to m-calpain, Lp82 truncated only the first five amino acids from the C-terminus of alphaA-crystallin. CONCLUSIONS: Under pathologic conditions of massive elevation of lens calcium found in young rodent lenses, overactivation of Lp82 and m-calpain leads to rapid truncation of crystallins at both common and unique cleavage sites, precipitation of truncated crystallins, and cataract. PMID- 10798665 TI - Transition metal-catalyzed oxidation of ascorbate in human cataract extracts: possible role of advanced glycation end products. AB - PURPOSE: With age, human lens crystallins become more pigmented, oxidized, modified by ascorbate oxidation and advanced glycation end products (AGEs), and bind copper. The hypothesis was tested that the major AGE and ascorbylation product in the human lens, N(epsilon)-carboxymethyl-L-lysine (CML), has an EDTA like structure, which may predispose it to bind redox active copper. METHODS: Young, old, and cataractous human lens protein fractions were glycated with ascorbic acid and tested for their ability to bind Cu(II) by atomic absorption spectroscopy and oxidize (14C1)-ascorbate by radiometric thin-layer chromatography method. AGEs were assayed by high-performance liquid chromatography (HPLC). CML-rich proteins were immunoprecipitated from young, old, and cataractous crystallins using affinity-purified CML antibody and tested for their ability to oxidize ascorbate and generate hydroxyl radicals in the presence of H2O2 using 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO) spin-trap and EPR spectroscopy. RESULTS: Ascorbate oxidizing activity at 24 hours of native crystallins was significantly increased in both the water soluble (WS; P < 0.001) and insoluble (WIS; P < 0.05) fractions from cataractous and normal lenses. The chelator DTPA completely prevented oxidation up to 24 hours of incubation but less effectively thereafter. Mean endogenous Cu content in pooled young, old, and cataract fractions increased from 0.016 to 0.026 nmol/mg protein, respectively, in WS (P < 0.05) and WIS (P < 0.001) fractions, and Cu(II) binding was 20% to 30% increased in cataractous versus old and young lenses in WS (P < 0.01) and WIS (P < 0.001) fractions. Mean levels of the AGEs, CML, and pentosidine were markedly elevated in WS and WIS fractions from cataractous versus old or young crystallins (20% to severalfold, P < 0.05 to P < 0.001). In a separate experiment, protein bound Fe was not elevated. Crystallins ascorbylated in vitro showed an increase in CML as well as Cu(II) binding. CML-rich proteins (immunoprecipitated from cataractous lenses) oxidized ascorbate approximately 4 times faster than similar proteins from young and old normal lenses (P < 0.01) and generated hydroxyl radicals in the presence of H2O2 and DMPO. CONCLUSIONS: The association between CML formation, copper binding, and generation of free radicals by cataractous lens crystallins can be duplicated by ascorbylation in vitro. These effects are only in part attributable to CML itself, and other modifications (AGEs, conformational changes) may participate in the process. A vicious cycle between AGE formation, lipoxidation, and metal binding may exist in the aging lens, suggesting that chelation therapy could be beneficial in delaying cataractogenesis. PMID- 10798666 TI - Erythrocyte sodium-lithium countertransport and proliferative diabetic retinopathy. AB - PURPOSE: To investigate whether elevated erythrocyte Na+/Li+ countertransport (Na+/Li+ CT) activity is present in patients with proliferative diabetic retinopathy (PDR). METHODS: The rate of Na+/Li+ CT activity assayed in 21 patients with type 1 diabetes mellitus (DM) presenting PDR was compared with 10 patients with nonproliferative retinopathy (NPDR) and with 11 patients with normal fundi. Twelve normal volunteers with no family history of hypertension were used as a control group. The albumin excretion rate was determined by nephelometry, and the glomerular filtration rate was measured by the plasma clearance of eidetic acid labeled with chromium-51. RESULTS: Patients with PDR showed higher diastolic blood pressure levels (mean +/- SD) compared with those with NPDR or normal fundi (95 +/- 13 versus 90 +/- 09 and 82 +/- 19 mm Hg, P = 0.02, respectively). The albumin excretion rate was higher [geometric mean (range)], and the glomerular filtration rate was lower (mean +/- SD) in patients with PDR than in those with NPDR or normal fundi [333 (2 to 5140) versus 32 (5.9 to 2200) and 6 (1.5 to 306) microg/min, P = 0.01, and 63 +/- 33 versus 99 +/- 37 and 93 +/- 43 ml/min, P = 0.02, respectively]. The mean Na+/Li+ CT in patients with PDR was significantly higher than in patients with NPDR or normal fundi and control group (0.46 +/-0.20 versus 0.32 +/- 0.12, 0.32 +/- 11, and 0.21 +/- 0.07 mM/L red blood cells (RBC)/h, respectively, P = 0.0001). In a multiple logistic regression analysis, with PDR as the dependent variable, Na+/Li+ CT (odds ratio [OR]: 4.7, confidence interval [CI]: 1.2-17.6, P = 0.02), diastolic blood pressure (OR, 3.4; CI, 1.3 to 9.6; P = 0.018), and glomerular filtration rate (OR, 5.1; CI, 1.6-17.7; P = 0.007) were the only variables that were maintained in the equation, indicating that they were the main determinants of PDR. CONCLUSIONS: Patients with type 1 DM and proliferative retinopathy have elevated erythrocyte Na+/Li+ CT. PMID- 10798667 TI - Intersession repeatability of macular thickness measurements with the Humphrey 2000 OCT. AB - PURPOSE: This study was designed to determine intersession repeatability of measurements of macular thickness made with a commercially available optical coherence tomography (OCT) system. The images that can be routinely acquired with the commercial instrument differ significantly in quality from the images in the literature, which have mostly been acquired on prototype systems. METHODS: Multiple OCT images of the nasal macula were obtained from the right eye during three independent measuring sessions, using the Humphrey 2000 OCT system (Humphrey, San Leandro, CA). Twenty-six volunteers with no history of ocular disease participated in this investigation. Eyes in all subjects were undilated during scan acquisition. Scans were horizontal, 3 mm long, and through the fovea. Five scans were used from each session, for a total of 15 scans per subject. Retinal boundaries were automatically detected using custom software. Statistical software was used to calculate intersession and intrasession repeatability. Manual correction was performed on the automatically detected boundaries, and a second analysis was performed using these boundaries. RESULTS: When no manual correction of boundaries was performed, there were no significant effects between different sessions (P = 0.529) or between different scans within the same session (P = 0.509). Average retinal thickness was found to be 274 +/- 17 microm for a 1 mm long region 0.75 mm from the fovea. Individual scan averages differed from overall patient averages by 0 +/- 4.3 microm (99% confidence interval, 11.2 microm). CONCLUSIONS: OCT measurements of macular thickness made with the Humphrey 2000 OCT system are repeatable over different sessions with an expected variation of less than 11 microm (99% confidence interval). PMID- 10798668 TI - Macular pigment optical density in a Southwestern sample. AB - PURPOSE: Increasing evidence implicates macular pigment in protecting the retina and retinal pigment epithelium from light-initiated oxidative damage. Little information, however, is available regarding "average" levels of macular pigment in the general population. This study was designed to assess macular pigment in a high-light environment and to determine what personal characteristics influence macular pigment density in that sample. METHODS: Macular pigment optical density was measured psychophysically using a 1 degree, 460-nm test stimulus. Personal data were collected using a questionnaire. RESULTS: 217 subjects (79 men, 138 women) were recruited from the Phoenix metropolitan area (age range = 17-92 years). The average macular pigment density was 0.22 +/- 0.13. There was a slight tendency for macular pigment density in this sample to decline with age (r = 0.14, P < 0.02). Average macular pigment density was significantly lower in women versus men (P < 0.05), lower in individuals with light-colored irises versus dark colored irises (P < 0.009), and lower in heavy smokers compared to light (P < 0.0045) and never (P < 0.034) smokers. CONCLUSIONS: Macular pigment density was lower than average levels obtained from the Northeast but similar to average values obtained in a recent study of adults recruited from Indianapolis. Consistent with past studies, MP density was 13% lower in women and 18% lower in individuals with light- versus dark-colored irises. The relation of smoking to macular pigment density was only significant for those current smokers who smoked more than 10 cigarettes per day (about a 25% reduction). The large number of individuals in this sample with low macular pigment density motivates the need for population-based assessment of the possibly poor nutritional state of the average American's retina. PMID- 10798669 TI - Retinal sensitivity to flicker modulation: reduced by early age-related maculopathy. AB - PURPOSE: To evaluate retinal, cone-mediated flicker sensitivity (CFS) in age related maculopathy (ARM) by quantifying response gain and threshold of the focal electroretinogram (FERG) to flicker modulation. METHODS: Nineteen patients with ARM (visual acuity > or =20/30) and 11 age-matched control subjects were examined. Twelve patients had less than 20 soft drusen in the macular region and no hyper-/hypopigmentation (early lesion), whereas seven had more than 20 soft drusen and/or focal hyper-/hypopigmentation (advanced lesion). Macular (18 degree ) FERGs were elicited by a sinusoidally flickering (41 Hz) uniform field (on a light-adapting background) whose modulation depth was varied between 16.5% and 94%. Amplitude and phase of the response's fundamental harmonic were measured. RESULTS: In both control subjects and patients with ARM, log FERG amplitude increased with log stimulus modulation depth with a straight line (power law) relation. However, the slope (or gain) of the function was, on average, steeper in control subjects than in patients with either early or advanced lesions. Mean FERG threshold, estimated from the value of the log modulation depth that yielded a criterion response, did not differ between control subjects and patients with early lesions but was increased (0.35 log units) compared with control subjects in those with advanced lesions. In both patient groups, but not in control subjects, mean FERG phase tended to delay with decreasing stimulus modulation depth. CONCLUSIONS: Retinal CFS losses can be detected in ARM by evaluating the FERG as a function of flicker modulation depth. Reduced response gain and phase delays, with normal thresholds, are associated with early lesions. Increased response thresholds, in addition to gain and phase abnormalities, may reflect more advanced lesions. Evaluating CFS by FERG may directly document different stages of macular dysfunction in ARM. PMID- 10798670 TI - Squalamine improves retinal neovascularization. AB - PURPOSE: Modalities for inhibiting neovascularization may be one avenue to the development of effective therapies for retinopathy. The effect of squalamine, an antiangiogenic amino sterol, on oxygen-induced retinopathy (OIR) was assessed in a mouse model. METHODS: OIR was induced in C57BL6 mice by a 5-day exposure to 75% oxygen from postnatal day (P)7 through P12. Squalamine (25 mg/kg, subcutaneous)treated animals received either daily doses for five days from P12 to P16 or one dose just after removal from oxygen on P12. Each set of animals was killed at P17 to P21. Retinopathy was assessed with a retinopathy scoring system evaluation of retinal wholemounts and by quantification of neovascular nuclei on retinal sections. RESULTS: Animals receiving 5 days of squalamine after a 5-day exposure to oxygen had total retinopathy scores (expressed as median score with 25th and 75th quartiles in parentheses) of 4(3, 5) versus oxygen-only-reared animals with scores of 8(7, 9; P < 0.001). Animals reared in room air and animals exposed to squalamine only had similar retinopathy scores: 1(1, 2) and 1(0, 2). Oxygen-reared animals receiving single-dose squalamine also showed improvement, with a median retinopathy score of 4(4, 6.75) versus oxygen-only-reared animals with median retinopathy score of 9(7, 10; P < 0.001). There was a decreased number of neovascular nuclei extending beyond the inner limiting membrane on retinal sections in animals treated with 5 days (P < 0.01) and 1 day (P < 0.001) of squalamine. CONCLUSIONS: Squalamine significantly improved retinopathy and may be a novel agent for effective treatment of ocular neovascularization. PMID- 10798671 TI - Dose dependency of canthaxanthin crystals in monkey retina and spatial distribution of its metabolites. AB - PURPOSE: To establish the threshold level of canthaxanthin crystals in the retina of cynomolgus monkeys. To correlate the spatial distribution of all-trans canthaxanthin and its metabolites with the grade of crystals. METHODS: Monkeys were orally administered 0, 0.2, 0.6, 1.8, 5.4, 16.2, and 48.6 mg/kg body wt canthaxanthin daily for 2.5 to 3 years. A second group of monkeys were administered 200 and 500 mg/kg body wt/d for 5 years. Ophthalmoscopy, electroretinography (ERG), retina and carotenoid analysis were performed as previously reported. RESULTS: Crystals in the retina periphery were observed by ophthalmoscopy preterminally only in the extreme high doses of 200 to 500 mg/kg body wt/d. There were no adverse effects on visual functions as measured by ERG. Crystals in the peripheral retina, and/or in the macula, were detected microscopically in all canthaxanthin treated groups except at the lowest dose of 0.2 mg/kg body wt/d. The grade of crystals increased up to a dose of 16.2 mg/kg body wt/d. Dose-dependent increases in canthaxanthin content also were noted in the retina, the liver, and in plasma. All-trans canthaxanthin was the major compound in the peripheral and paracentral retina of very highly dosed animals, where its concentration correlated largely with the grade of inclusions. In the macula, 4'-OH-echinenone was the dominant canthaxanthin metabolite. CONCLUSIONS: The grade of crystals in monkey retinas was dose dependent with a threshold level at 0.6 mg canthaxanthin/kg body wt/d. It correlated in the retinal periphery with the concentrations of all-trans-canthaxanthin and in the macula with its metabolites. PMID- 10798672 TI - Intravitreal toxicology and duration of efficacy of a novel antiviral lipid prodrug of ganciclovir in liposome formulation. AB - PURPOSE: To evaluate the intraocular safety and antiviral treatment efficacy of the sustained lipid prodrug of ganciclovir, 1-O-hexadecylpropanediol-3-phospho ganciclovir (HDP-P-GCV), as an intravitreal injectable drug system for viral retinitis. METHODS: HDP-P-GCV was synthesized by coupling 1-O-hexadecyl propanediol-3-phosphate to either free hydroxyl of ganciclovir in pyridine with dicyclohexylcarbodiimide as catalyst. The compound was formulated into liposomes. The antiviral activity was assessed by DNA reduction in vitro, and intraocular safety was assessed by ophthalmoscopy, electrophysiology, and histology after intravitreal injections, with resultant intravitreal concentrations of 0.2, 0.632, 1.12, and 2 mM. The treatment efficacy was evaluated by simultaneous intravitreal injection of HDP-P-GCV and herpes simplex virus type 1 (HSV-1) or by intravitreal injection of HDP-P-GCV at various times before HSV-1 intravitreal inoculation. Retinitis was scored with ophthalmoscopy and compared with controls. RESULTS: In vitro, the IC50 of HDP-P-GCV against HSV-1 and human cytomegalovirus (HCMV) infected cells was 0.02 and 0.6 microM, respectively. In rabbits in vivo, HDP-P-GCV dispersed evenly and maintained a good vitreous clarity at all doses except 2 mM final intravitreal concentration. Although cataracts were observed in some eyes at the higher doses, they were not observed in eyes with 0.2 mM final intravitreal concentration. No other indications of ocular toxicity were observed. Intravitreal injection of HDP-P-GCV with resultant 0.2 mM intravitreal concentration in the HSV-1 retinitis rabbit model demonstrated a complete protection of the retina with the simultaneous treatment strategy and a 4 (P = 0.03) to 6-(P = 0.058) week significant protection of retina with the pretreatment strategies when compared with ganciclovir or blank liposome controls. CONCLUSIONS: In the rabbit model of HSV-1 retinitis HDP-P-GCV acts as a long-lasting intravitreal injectable anti-CMV or anti-HSV compound. This self assembling liposome system could be applicable for many compounds available for intraocular diseases. PMID- 10798673 TI - Differential expression of neuroendocrine-specific protein in form-deprived chick eyes. AB - PURPOSE: To identify genes that are highly expressed in form-deprived retina retinal pigment epithelium-choroid tissues. Neuroendocrine-specific proteins were found to be highly expressed. METHODS: mRNAs enriched in retina-retinal pigment epithelium-choroid tissues from 3-, 7-, and 14-day form-deprived chick eyes were isolated by differential display technique with cDNA library screening. Neuroendocrine-specific protein A and C were cloned in control and form-deprived eyes. mRNA and protein levels, with respective regional localizations, were examined by Northern blot, Western blot, and immunohistochemical analyses, respectively. RESULTS: The isolated clone included an insert with a sequence homologous to both chick neuroendocrine-specific proteins A and C. The increases in mRNA and protein levels were confirmed by Northern and Western blot analyses, respectively. Immunohistochemical localization of neuroendocrine-specific proteins A and C was detected in the layer of photoreceptor inner segments, presumably in the cone cells. Northern blot analysis using negative lenses showed that levels of neuroendocrine-specific protein A and C mRNAs were not altered using negative lenses. CONCLUSIONS: The expression of both neuroendocrine specific proteins A and C mRNAs in cone photoreceptor cells was upregulated within 14 days of form deprivation, but not in response to negative spectacle lenses. These data suggest that the increase in induction of neuroendocrine specific proteins is not a secondary consequence of ocular elongation or myopic refraction. Induction of neuroendocrine-specific proteins in form-deprived eyes may be causally related to the development of myopia or may be an unrelated effect of form deprivation. PMID- 10798674 TI - Muller cell outgrowth after retinal detachment: association with cone photoreceptors. AB - PURPOSE: Subretinal gliosis is a relatively common occurrence after retinal reattachment. Because Muller cell processes only intermittently penetrate the outer limiting membrane (OLM) beneath extensive detachments, this study was conducted to determine whether this was preferentially associated with rod or cone photoreceptors. METHODS: Cat retinas were detached from the retinal pigment epithelium and 3 days later were fixed in 4% paraformaldehyde, embedded in 5% agarose, sectioned at 100 microm, and processed for standard immuohistochemistry. The retinas were double labeled with either anti-vimentin and anti-long/medium wavelength-sensitive (anti-L/M) cone opsin or anti-glial fibrillary acidic protein (GFAP) and biotinylated peanut agglutinin (PNA). RESULT: The hypertrophy of Muller cells was readily traced using antibodies to vimentin and GFAP. When labeling with these antibodies was combined with labeling by either antibodies to cone opsins or biotinylated PNA, a consistent relationship was observed between the Muller cell processes growing through the OLM and cone photoreceptors. CONCLUSIONS. The growth of Muller cell processes into the subretinal space forms a fibrotic layer that completely inhibits the regeneration of outer segments. The current results show that there appears to be a highly specific interaction between growing Muller cell processes and cone photoreceptors during the earliest phase in this process. PMID- 10798675 TI - Characterization of rhodopsin mis-sorting and constitutive activation in a transgenic rat model of retinitis pigmentosa. AB - PURPOSE: To determine the extent to which rhodopsin mis-sorting and constitutive activation of the phototransduction cascade contribute to retinal degeneration in a transgenic rat model of retinitis pigmentosa. METHODS: Retinas from transgenic rats expressing truncated rhodopsin (Ser334ter) were examined by light and electron microscopic immunocytochemistry at several time points. Retinal degeneration in transgenic rats raised in darkness was evaluated by quantification of outer nuclear layer thickness and by electroretinography. RESULTS: Mutant rhodopsin was found at inappropriately high levels in the plasma membrane and cytoplasm of Ser334ter rat photoreceptors. When the cell death rate was high this mis-sorting was severe, but mis-sorting attenuated greatly at later stages of degeneration, as the cell death rate decreased. The distributions of two other outer segment proteins (the cGMP-gated channel and peripherin) were examined and found to be sorted normally within the photoreceptors of these rats. Raising Ser334ter transgenic rats in darkness resulted in minimal rescue from retinal degeneration. CONCLUSIONS: Because dark rearing Ser334ter rats results in little rescue, it is concluded that constitutive activation of the phototransduction cascade does not contribute significantly to photoreceptor cell death in this rat model. The nature of the rhodopsin sorting defect and the correlation between the severity of mis-sorting and rate of cell death indicate that truncated rhodopsin may cause apoptosis by interfering with normal cellular machinery in the post-Golgi transport pathway or in the plasma membrane. PMID- 10798676 TI - Glucose-specific regulation of aldose reductase in human retinal pigment epithelial cells in vitro. AB - PURPOSE: To test the hypothesis that pathophysiological levels of glucose regulate aldose reductase (AR2) gene expression, protein production, and activity in human retinal pigment epithelial (RPE) cells in vitro. METHODS: Primary cultures of human RPE cells were grown for up to 72 hours in media supplemented with various concentrations of glucose (5, 20, or 75 mM), or in 5 mM glucose containing media supplemented with one of the following: galactose, the transported but nonmetabolized glucose analogue 3-O-methylglucose (3-OMG), or the impermeant hexitol mannitol-so that the final hexose concentrations were equimolar to those of the various glucose concentrations used. Changes in the transcript levels for AR2 mRNA, AR2 protein content, and AR2 enzyme activity were determined. RPE glucose utilization and lactate production were determined in media containing 5 and 20 mM glucose. RESULTS: Glucose utilization and lactate production increased 4.8-fold and 4.4-fold, respectively, when RPE cells were grown in media containing 20 mM versus 5 mM glucose. Glucose was more effective than any other hexose in the induction of AR2 mRNA or increased AR2 protein expression. When RPE cells were grown in media containing 20 mM mannitol, 3-OMG, or galactose they had lower levels of AR2 mRNA expression than when cells were grown in medium containing 5 mM glucose. RPE cells grown in medium supplemented with 20 or 75 mM galactose did not show a greater increase in AR2 protein expression than cells grown in medium containing 5 mM glucose. Hyperosmotic induction of AR2 mRNA was the same in medium containing 75 mM glucose or 75 mM mannitol, but was at least 50% lower when RPE cells were grown in 75 mM galactose or 3-OMG. CONCLUSIONS. These data indicate that elevations in ambient glucose result in greater metabolism of glucose through glycolysis and polyol metabolism. Induction of AR2 was greatest when RPE cells were grown in pathophysiological concentrations of glucose. Hyperosmolar stress is not a necessary determinant of AR2 mRNA, AR2 protein, or AR2 protein activity in cells that form the outer blood retinal barrier. Increased facilitative glucose transport or glucose metabolism appears to be requisite for glucose-specific and nonosmotic regulation of AR2 in the RPE cell in vitro. PMID- 10798677 TI - Quantitative MR imaging study of intravitreal sustained release of VEGF in rabbits. AB - PURPOSE: To determine whether sustained elevation of vascular endothelial growth factor (VEGF) in the vitreous cavity causes retinal hyperpermeability [blood retinal barrier (BRB) breakdown] before the development of retinal neovascularization (NV) and to document the kinetics of the integrity of BRB breakdown versus time. METHODS: Poly(L-lactide-co-glycolide)based devices loaded with VEGF were implanted intravitreally in rabbit eyes. Contrast-enhanced magnetic resonance imaging (MRI) methods were used to identify and quantitate the retinal permeability at various time points after implantation. This was done with the newly developed MR tracer AngioMARK (Epix Medical, Boston, MA). After the MRI measurements, fundus photography and fluorescein angiography (FA) also were performed on the same set of animals. RESULTS: At 3 days after implantation, the MR images showed a significant retinal leakage into the vitreous cavity (BRB breakdown) of the VEGF-implanted eyes. To quantitate this leakage, the permeability surface area product (PS) was measured. At 3 days, the mean PS product was 1.25 +/-0.25 x 10(-5) cm3/min. Based on the VEGF in vitro release study, this 3-day BRB breakdown corresponded to a total sustained release of 7.42 +/- 0.54 microg/ml of VEGF. The fundus and FA photographs of these VEGF-implanted eyes taken at 4 days after implantation also showed a considerable level of retinal vascular dilation and tortuosity. By 12 days after implantation, the mean PS product decreased to 5.83 +/- 1.38 x 10(-6) cm3/min. However, the retinal NV was observed only after the second week after implantation. By this time, a total of 10.70 +/- 0.92 microg/ml of VEGF was released in a sustained fashion. Also, after the retinal NV development, retinal detachment also was observed. The control eyes, however, which were implanted with blank devices, remained unchanged and normal during the entire course of this study (PS = 5.57 +/- 0.66 x 10(-7) cm3/min). CONCLUSIONS. The findings indicate that sustained delivery of elevated amounts of VEGF in the vitreous cavity induces a BRB breakdown even earlier than 3 days after implantation. This was achieved after a total sustained release of 7.42 +/- 0.54 microg/ml of VEGF. This retinal leakage regressed by more than half by the time the retinal NV developed. Furthermore, a retinal detachment occurred after this retinal NV. These results are similar to proliferative diabetic retinopathy (PDR). The sustained elevation of VEGF in the vitreous cavity of rabbit eyes is potentially a good model to test VEGF antagonists to treat or prevent PDR in humans. The quantifiable change of BRB breakdown by the contrast-enhanced MRI method is ideal to assess the therapeutic intervention in vivo without killing the animal and may prove to be clinically useful in humans. PMID- 10798678 TI - An attempt to detect glaucomatous damage to the inner retina with the multifocal ERG. AB - PURPOSE: To detect glaucomatous damage to the inner retina using the multifocal electroretinogram (mERG). METHODS: The stimulus array consisted of 103 hexagons with a mean luminance of 100 cd/m2 and a contrast of 50%. The mERG was recorded from 13 control subjects, 18 patients with open-angle glaucoma (OAG), 4 glaucoma suspects, and one patient with ischemic optic neuropathy (ION). Individual responses, as well as responses summed within quadrants or across the entire array, were measured in a number of ways. Humphrey visual fields were obtained for all patients, and the mean total deviation (MD) values for the 18 patients with OAG ranged from -2.2 to -18.2 with a mean (SD) of -7.3 (4.5). RESULTS: The mERG measure that best discriminated between the patients and the control subjects was the ratio of the amplitude at 8 msec after the peak response to the amplitude at the peak. Although the value of this ratio fell below the median of the control group for 16 of the 18 OAG patients, only 6 of these patients had ratios that fell below the normal range. Other measures of first- and second order kernels did not do as well. Both within and across patients, the correlation between local field loss and the mERG ratio measure was poor. Furthermore, although in some patients the mERG waveform is clearly different from normal, in other patients (including the patient with ION) the waveform approximates the normal even in visual field areas with substantial sensitivity loss. CONCLUSIONS: Because glaucomatous damage is known to affect the ganglion cell axon, these data suggest that damage to ganglion cell axons is not a sufficient condition to produce changes in the mERG as measured here and that in patients with clear changes in mERG waveforms, these changes do not appear to be well localized and local waveforms are poorly correlated with local changes in field sensitivity. PMID- 10798679 TI - An interocular comparison of the multifocal VEP: a possible technique for detecting local damage to the optic nerve. AB - PURPOSE: To develop a quantitative measure of local damage to the ganglion cells/optic nerve based on an interocular comparison of multifocal visual evoked potentials (mVEP). METHODS: Multifocal VEPs were recorded from both eyes of six normal subjects and four patients; each eye was stimulated separately. Two of the patients had glaucoma, one had ischemic optic neuropathy, and one had unilateral optic neuritis. All four patients had considerably more damage in one eye than in the other, as indicated by their Humphrey visual fields. The multi-input procedure of Sutter was used to obtain 60 VEP responses to a scaled checkerboard pattern. The amplitude in each response was obtained using a root mean square measure of response magnitude. For each of the 60 pairs of responses, a ratio between the amplitude of the responses from the two eyes was obtained as a measure of the relative health of one eye compared with the other. The mean and SD of this ratio measure for the control group were used to specify confidence intervals for each of the 60 locations. All patients had Humphrey 24-2 visual fields performed. To allow a comparison of the mVEPs to the visual fields, a procedure was developed for displaying the results of both tests on a common set of coordinates. RESULTS: Except for a small interocular difference in timing attributable to nasotemporal retinal differences, the pairs of mVEP responses from the two eyes of the control subjects were essentially identical. Many of the pairs of responses from the patients were significantly different. In general, there was reasonably good agreement with the Humphrey 24-2 visual field data. Although some regions with visual field defects were not detected in the mVEP due to small responses from the better eye, other abnormalities were detected that were hard to discern in the visual fields. CONCLUSIONS: Local monocular damage to the ganglion cell/optic nerve can be quantitatively measured by an interocular comparison of the mVEP. PMID- 10798680 TI - The development of scotopic sensitivity. AB - PURPOSE. Test the hypothesis that the developmental increases in rod photoreceptor sensitivity and rod-mediated visual sensitivity at 10 degrees, 20 degrees , and 30 degrees eccentric are concurrent. It is known that maturation of the parafoveal (10 degrees eccentric) rod outer segments and visual sensitivity is delayed compared to that at 30 degrees eccentric. METHODS: Rod isolated electroretinographic (ERG) responses to full-field stimuli were obtained from dark-adapted subjects (n = 71), ranging in age from early infancy through middle age. Rod photoreceptor sensitivity was calculated by fitting a model of the activation of phototransduction to the a-wave response. Rod driven b-wave sensitivity was calculated from stimulus-response functions. A logistic growth model was used to summarize the developmental increases in sensitivity of the rod photoreceptors and the b-wave. Previously reported dark-adapted, rod-mediated visual sensitivities at 10 degrees , 20 degrees, and 30 degrees eccentric, obtained using preferential looking procedures, were reanalyzed using the logistic growth model. RESULTS: The logistic growth model accounted for 57% to 85% of the variance of each sensitivity parameter with age in normal subjects. The shape of the growth curve and the age at which sensitivity reaches 50% of the adult value is similar (10.0-13.5 weeks) for the rods, the b-wave, and peripheral visual sensitivity, but is significantly older, 19.5 weeks, for rod-mediated parafoveal visual sensitivity. CONCLUSIONS: Rod photoreceptor sensitivity and peripheral, rod-mediated visual sensitivity develop concurrently. A parsimonious explanation is that rod photoreceptor sensitivity determines dark-adapted, rod mediated visual sensitivity during development. PMID- 10798681 TI - Waveform changes of the first-order multifocal electroretinogram in patients with glaucoma. AB - PURPOSE: To investigate the relationship between the components of the first order multifocal electroretinogram (M-ERG) and glaucomatous visual field loss. METHODS: Twenty-six eyes of 14 patients with primary open-angle glaucoma (POAG) were evaluated with the M-ERG techniques. Twenty-six eyes of 26 normal subjects also were tested as control subjects. To record the M-ERG, a stimulus matrix of 103 scaled hexagonal elements was displayed on a monitor driven at a 75-Hz frame rate according to a binary m-sequence. The M-ERG responses were averaged in each quadrant of the stimulus field and the peak-to-trough amplitudes and peak implicit times of the first trough (N1), the first peak (P1), and the second trough (N2) of the M-ERG were compared with the mean sensitivity values (dB) of the corresponding quadrant of the Humphrey static perimetric field. RESULTS: The changes in the peak latencies of P1 and N2 in the POAG group were small but significant compared with those in the normal group (P < 0.01). However, no significant differences in the amplitudes of (P1-N1) and (P1-N2) between the two groups were found. Significant negative correlations between the peak implicit times of N1, P1, and N2 and the mean sensitivity values (dB) of static perimetry were observed. The correlation coefficients were -0.20 (P < 0.05) for the N1, 0.41 (P < 0.001) for the P1, and -0.59 (P < 0.001) for the N2. No significant correlations were observed between the amplitudes (P1-N1 and P1-N2) and the mean sensitivity values. CONCLUSIONS: The present study findings suggest that the peak implicit times, but not the amplitudes, of the M-ERG increase as the glaucomatous visual field deteriorates. The amplitudes of the M-ERG did not decrease as the glaucomatous optic nerve dysfunction progressed. PMID- 10798682 TI - Medical physicists should position themselves as institutional resources in expanding areas such as healthcare informatics and information networking. PMID- 10798683 TI - Recommendations of the American Association of Physicists in Medicine on 103Pd interstitial source calibration and dosimetry: implications for dose specification and prescription. AB - The National Institute of Standards and Technology (NIST) introduced a national standard for air kerma strength of the ThreaSeed Model 200 103Pd source (the only 103Pd seed available until 1999) in early 1999. Correct implementation of the NIST-99 standard requires the use of dose rate constants normalized to this same standard. Prior to the availability of this standard, the vendor's calibration procedure consisted of intercomparing Model 200 seeds with a 109Cd source with a NIST-traceable activity calibration. The AAPM undertook a comprehensive review of 103Pd source dosimetry including (i) comparison of the vendor and NIST-99 calibration standards; (ii) comparison of original Task Group 43 dosimetry parameters with more recent studies; (iii) evaluation of the vendor's calibration history; and (iv) evaluation of administered-to-prescribed dose ratios from the introduction of 103Pd sources in 1987 to the present. This review indicates that for a prescribed dose of 115 Gy, the administered doses were (a) 124 Gy for the period 1988-1997 and (b) 135 Gy for the period 1997-1999. The AAPM recommends that the following three steps should be undertaken concurrently to implement correctly the 1999 dosimetry data and NIST-99 standard for 103Pd source: (1) the vendor should provide calibrations in terms of air kerma strength traceable to NIST-99 standard, (2) the medical physicist should update the treatment planning system with properly normalized (to NIST-99) dosimetry parameters for the selected 103Pd source model, and (3) the radiation oncologist in collaboration with the medical physicist should decide which clinical experience they wish to duplicate; the one prior to 1997 or the one from 1997 to 1999. If the intent is to duplicate the experience prior to 1997, which is backed by the long-term follow-up and published outcome studies, then the prior prescriptions of 115 Gy should be replaced by 124 Gy to duplicate that experience. PMID- 10798685 TI - Measurement of dose-rate constant for 103Pd seeds with air kerma strength calibration based upon a primary national standard. AB - Recent developments in the past two years require a significant change in the dosimetry of 103Pd brachytherapy sources (Theraseed model 200, manufactured by Theragenics Corp., Atlanta, GA). Since their introduction in 1987, the air kerma strength of 103Pd sources for interstitial brachytherapy has been determined using a system of apparent activity measurement based upon the measurement of photon fluence at a reference distance along the transverse axis of the source free in air, using a NaI (T1) scintillation detector at the manufacturer's facilities. This detection system has been calibrated against a National Institute of Standards and Technology (NIST)-traceable activity standard of a 109Cd source. This system produced a highly consistent standard (within +/-2%) for over 12 years, with the exception of the last 109Cd source change in September 1997, which resulted in a change of 9% from the original 1987 standard. The second major development affecting 103Pd dosimetry is that on 13 January 1999 a primary national standard for the air kerma strength of 103Pd seeds was developed by NIST. This primary standard is based upon an absolute measurement of air kerma rate free in air at a reference distance from the source along its transverse axis using a wide angle free air chamber (WAFAC). In order to implement this new standard for the calibration of source strength in clinical dosimetry for interstitial implants, it is necessary to measure the dose-rate constant for the 103Pd seeds using a calibration of source strength based on the NIST 99 standard. In this work, a measurement of the dose-rate constant using lithium fluoride (LiF) thermoluminescent dosimeters (TLDs) in a water equivalent solid phantom is reported. The measured value of this constant is 0.65 +/- 0.05 cGy h(-1) U(-1), where the unit air kerma strength is 1 U = 1 cGy h(-1) cm2 = 1 microGy h(-1) m2, and is directly traceable to the NIST 99 standard. The implementation of the NIST 99 standard for 103Pd should be accompanied by a simultaneous adoption of the new dose-rate constant reported here. No changes in radial dose function, anisotropy function, anisotropy factor, and geometry function are needed. However, a change in prescribed dose may be necessary to deliver the same physical dose as before. PMID- 10798684 TI - Monte Carlo modeling of the transverse-axis dose distribution of the model 200 103Pd interstitial brachytherapy source. AB - This study presents the first theoretical analysis of the absolute dose-rate distribution about the Model 200 103Pd interstitial brachytherapy source. Monte Carlo photon-transport (MCPT) simulation techniques have been used to evaluate the transverse-axis dose-rate distribution of the Model 200 source as a function of thickness of the Pd metal coating (containing the 103Pd) plated onto the surfaces of right cylindrical graphite pellets contained within the seed. The dose-rate constant, A, was realistically estimated by simulating the wide-angle, free-air chamber (WAFAC) calibration geometry. The WAFAC is the experimental realization of NIST's (National Institute of Standards and Technology) recently implemented primary standard of air-kerma strength (S(K)). Our results show that polar angle- and distance-dependent oblique filtration and shielding effects induce significant and unexpected photon fluence anisotropy near the transverse axis and inverse square law deviations at typical calibration distances. Any source consisting of radioactivity deposited on a highly attenuating surface with sharp edges may exhibit such effects. In the case of the Model 200 seed, the Pd metal thickness does not significantly influence the relative dose distribution in water at distances less than 5 cm, but does make A sensitive to the S(K) measurement geometry. Fortunately, the WAFAC averages fluence over a sufficiently large aperture that the resultant A, 0.68 +/- 0.02 cGy x h(-1) x U(-1), is almost independent of Pd metal layer thickness and in close agreement with recent measurements and calculations. This value is 20% higher than that of the renormalized Task Group 43 A value. PMID- 10798686 TI - Technical note: improved analytical fit to the TG-43 radial dose function, g(r). AB - An improved analytical fit for the TG-43 radial dose function, g(r), is presented using a modified sigmoid function. Fit coefficients and graphs are provided for comparison with the polynomial fits given in TG-43. PMID- 10798687 TI - A high-precision, high-resolution and fast dosimetry system for beta sources applied in cardiovascular brachytherapy. AB - A fast dosimetry system based on plastic scintillator detectors has been developed which allows three-dimensional measurement of the radiation field in water of beta-sources appropriate for application in cardiovascular brachytherapy. This system fulfills the AAPM Task Group 60 recommendations for dosimetry of cardiovascular brachytherapy sources. To demonstrate the use of the system, measurements have been performed with an 90Y-wire source. The dose distribution was determined with a spatial resolution of better than 0.2 mm, with only a few minutes needed per scan. The scintillator dosemeter was absolutely calibrated in terms of absorbed dose to water with a precision of +/-7.5%. The relative precision achievable is +/-2.5%. The response of the system is linear within +/-2% for dose rates from 0.5 mGy s(-1) to 500 mGy s(-1). PMID- 10798688 TI - Dosimetry of a W-188/Re-188 beta line source for endovascular brachytherapy. AB - PURPOSE: The objective was to determine the dosimetry of a potential endovascular brachytherapy source consisting of a coiled tungsten wire mounted on the distal end of a drive wire and neutron-activated to contain the parent-daughter nuclides tungsten-188 (188W) and rhenium-188 (188Re). METHODS: A coiled tungsten wire 40 mm in length was neutron-activated by double-neutron capture for 78 hours at 1.9 x 10(15) h/cm2/s to contain 925 MBq (25 mCi) of 188W/188Re in equilibrium. The dose-fall off from this source was determined using three independent methods: (a) Thermoluminescence dosimetry with small LiF-100 rods, (b) Gafchromic film dosimetry, and (c) Bang gel dosimetry. In addition, a Monte Carlo simulation was performed to compute the beta-dose. RESULTS: Each of the three measurement methods recorded similar values for the dose fall-off within the distances useful for endovascular brachytherapy. The Monte Carlo calculations closely approximated the measured results in the treatment range between 1 and 3 mm and may thus be useful for evaluating changing geometries in the development of catheters and source setups. A 2 min restenosis treatment delivering 20 Gy at a radius of 2 mm would require a source of 1384.8 MBq/cm (37.4 mCi/cm). CONCLUSIONS: The dose distribution from a 188W/188Re source is similar to that of a 90Y-source. An added advantage of the 188W/188Re source is that it can be used for at least two months and still provides fast treatment times because of the parent isotope's half-life of 69 days. The additional gamma emission from the source is too small to impose a serious radiological hazard. The high atomic number and density of the source material allows direct fluoroscopic imaging without additional markers. PMID- 10798689 TI - Electronic portal imaging with an avalanche-multiplication-based video camera. AB - The aim of this study is to investigate the degree to which the imaging quality of an existing (video-based) electronic portal imaging device (EPID) system may be improved by using an avalanche-multiplication-based video camera (called the avalanche-gain method). Due to avalanche multiplication in the target of the video camera tube, the new camera can be made up to several hundred times more sensitive than a camera using a conventional video (e.g., Saticon) tube. As a result, the camera noise which limits the performance of current video-based EPIDs should be overwhelmed and made negligible. The detective quantum efficiency (DQE) of an EPID using the avalanche-gain method has been measured with 6 MV and 18 MV beams obtained using a linear accelerator. It is shown that the camera noise is indeed much smaller than quantum noise and that the DQE of the system is significantly increased compared to conventional video-based EPIDs. Variation of DQE of the avalanche-gain video portal system with a change of demagnification was also investigated. It has been shown that the improvement of optical coupling has less effect in this system than that in a conventional video-based EPID system. PMID- 10798690 TI - Treatment planning algorithm corrections accounting for random setup uncertainties in fractionated stereotactic radiotherapy. AB - A number of relocatable head fixation systems have become commercially available or developed in-house to perform fractionated stereotactic radiotherapy (SRT) treatment. The uncertainty usually quoted for the target repositioning in SRT is over 2 mm, more than twice that of stereotactic radiosurgery (SRS) systems. This setup uncertainty is usually accounted for at treatment planning by outlining extra target margins to form the planning target volume (PTV). It was, however, shown by Lo et al. [Int. J. Radiat. Oncol., Biol., Phys. 34, 1113-1119 (1996)] that these extra margins partly offset the radiobiological advantages of SRT. The present paper considers dose calculations in SRT and shows that the dose predictions could be made at least as accurate as in SRS with no extra margins required. It is shown that the dose distribution from SRT can be calculated using the same algorithms as in SRS, with the measured off-axis ratios (OARs) replaced by "effective" OARs. These are obtained by convolving the probability density distribution of the isocenter positions (assumed to be normal) and the original OARs. An additional output correction factor has also been introduced accounting for the isocenter dose reduction (2.4% for a 7 mm collimator) due to the OARs "blurring." Another correction factor accommodates for the reduced (by 1% for 6 MV beam) dose rate at the isocenter due to x-ray absorption in the relocatable mask. Mean dose profiles and the standard deviations of the dose (STD) were obtained through simulating SRT treatment by a combination of normally distributed isocenters. These dose distributions were compared with those calculated using the convolution approach. Agreement of the dose distributions was within 1%. Since standard deviation reduces with the number of fractions, N, as STD/square root(N), the planning predictions in fractionated stereotactic radiotherapy can be made more accurate than in SRS by increasing N and using "effective" OARs along with corrected dose output. PMID- 10798691 TI - Optimization of inverse treatment planning using a fuzzy weight function. AB - A fuzzy approach has been applied to inverse treatment planning optimization in radiation therapy. The proposed inverse-planning algorithm optimizes both the intensity-modulated beam (IMB) and the normal tissue prescription. In the IMB optimization, we developed a fast-monotonic-descent (FMD) method that has the property of fast and monotonic convergence to the minimum for a constrained quadratic objective function. In addition, a fuzzy weight function is employed to express the vague knowledge about the importance of matching the calculated dose to the prescribed dose in the normal tissue. Then, a validity function is established to optimize the normal tissue prescription. The performance of this new fuzzy prescription algorithm has been compared to that based on hard prescription methods for two treatment geometries. The FMD method presented here both provides a full-analytical solution to the optimization of intensity modulated beams, and guarantees fast and monotonic convergence to the minimum. It has been shown that the fuzzy inverse planning technique is capable of achieving an optimal balance between the objective of matching the calculated dose to the prescribed dose for the target volume and the objective of minimizing the normal tissue dose. PMID- 10798692 TI - Algorithms and functionality of an intensity modulated radiotherapy optimization system. AB - The main purpose of this paper is to describe formalisms, algorithms, and certain unique features of a system for optimization of intensity modulated radiotherapy (IMRT). The system is coupled to a commercial treatment planning system with an accurate dose calculation engine based on the kernel superposition algorithm. The system was designed for use for research as well as for routine clinical practice. It employs dose- and dose-volume-based objective functions. The system can optimize IMRT plans with multiple target volumes simultaneously. Each target volume may be assigned a different prescription dose with constraints on either underdosing, or overdosing, or both. For organs at risk more than one constraint may be applied. This feature allows simultaneous treatment of primary, regional disease and electively treated nodes. The system allows specification of constraints on logical combinations of anatomic structures, such as a region of overlap between the prostate planning target volume and rectum or the volume of lung excluding the tumor. The optimization may also be performed on plans which, in addition to intensity-modulated beams, include other modalities such as non IMRT photon and electron beams and brachytherapy sources. The various features of the system are illustrated with one phantom example and two clinical examples: a brain stereotactic radiosurgery case and a nasopharynx case. In the cylindrical phantom example, the use of the system for overlap regions is demonstrated. The brain stereotactic radiosurgery example shows the improvement of IMRT plans over the conventional arcs based plan and the three-dimensional conformal plan with multiple fixed gantry angles and demonstrates the application of our system to cases where small grid sizes are important. The nasopharynx example shows the potential of IMRT to simultaneously treat large and boost fields. It also illustrates the power of IMRT to protect normal anatomic structures for highly complex situations and the efficiency in planning and delivery achievable with IMRT. The overall IMRT planning time is typically less than 2 h on a Sun Ultrasparc workstation, most of which is spent in repeated computation of dose distributions. PMID- 10798693 TI - A method for achieving variable widths of the spread-out Bragg peak using a ridge filter. AB - A method for designing a variable-SOBP (spread-out Bragg peak) ridge filter has been proposed. First, ridge filter parameters are determined by using a Monte Carlo calculation followed by a fast two-step iterative optimization. Then, tilting the ridge filter results in continuous variation of the SOBP width. Monte Carlo calculations show that depth dose uniformity changes from +/- 1.3% to +/- 1.6% for SOBP widths ranging from 10.3 cm to 14.5 cm. Advantages of the proposed tilting ridge filter include a capability of continuous SOBP variation and cost effective installation for a given SOBP width range. PMID- 10798694 TI - Ridge filter design and optimization for the broad-beam three-dimensional irradiation system for heavy-ion radiotherapy. AB - The broad-beam three-dimensional irradiation system under development at National Institute of Radiological Sciences (NIRS) requires a small ridge filter to spread the initially monoenergetic heavy-ion beam to a small spread-out Bragg peak (SOBP). A large SOBP covering the target volume is then achieved by a superposition of differently weighted and displaced small SOBPs. Two approaches were studied for the definition of a suitable ridge filter and experimental verifications were performed. Both approaches show a good agreement between the calculated and measured dose and lead to a good homogeneity of the biological dose in the target. However, the ridge filter design that produces a Gaussian shaped spectrum of the particle ranges was found to be more robust to small errors and uncertainties in the beam application. Furthermore, an optimization procedure for two fields was applied to compensate for the missing dose from the fragmentation tail for the case of a simple-geometry target. The optimized biological dose distributions show that a very good homogeneity is achievable in the target. PMID- 10798695 TI - Quality management of medical physics issues at the German heavy ion therapy project. AB - For the commissioning and operation of the German Heavy Ion Therapy Project a quality assurance program was developed and successfully applied. The complete radiotherapy process using heavy ions was carefully analyzed and divided into three areas related to beam delivery and control, safety-interlock system and medical physics issues. In this paper, the medical physics issues are addressed. Since the irradiation with heavy ions is a nonstandard modality, new concepts and ion specific tests were developed. As far as possible, national and international standard specifications for radiotherapy were adopted. For each aspect, a performance characteristic and a corresponding acceptance test were introduced. In addition, test characteristics for the constancy tests were established. For all tests, intervention thresholds and test frequencies were specified. Using the described protocol of acceptance tests, the commissioning was passed successfully. The heavy ion irradiation facility was approved by the governmental authorities on the basis of these test results. During clinical operation, constancy tests are performed at the beginning of each treatment period, in order to maintain the quality found during the acceptance tests. Up to now, 48 patients have been treated within 6 treatment periods of 4 weeks each. The concepts used and the tests developed for the quality assurance program may serve as an example of how to introduce systematically a quality assurance program for a new treatment modality. PMID- 10798696 TI - Modeling photon output caused by backscattered radiation into the monitor chamber from collimator jaws using a Monte Carlo technique. AB - Dose per monitor unit in photon fields generated by clinical linear accelerators can be affected by the backscattered radiation into the monitor chamber from collimator jaws. Thus, it is necessary to account for the backscattered radiation in computing monitor unit setting for a treatment field. In this work, we investigated effects of the backscatter from collimator jaws based on Monte Carlo simulations of a clinical linear accelerator. The backscattered radiation scored within the monitor chamber was identified as originating either from the upper jaws (Y jaws), or from the lower jaws (X jaws). From the results of Monte Carlo simulations, ratios of the monitor-chamber-scored dose caused by the backscatter to the dose caused by the forward radiation, R(x,y), were modeled as functions of the individual X and Y jaw positions. The amount of the backscattered radiation for any field setting was then computed as a compound contribution from both the X and Y jaws. The dose ratios of R(x,y) were then used to calculate the change in photon output caused by the backscatter, Scb(x,y). Results of these calculations were compared with available measured data based on counting the electron pulses or charge from the electron target of an accelerator. Data from this study showed that the backscattered radiation contributes approximately 3% to the monitor chamber-scored dose. A majority of the backscattered radiation comes from the upper jaws, which are located closer to the monitor chamber. The amount of the backscatter decreases approximately in a linear fashion with the jaw opening. This results in about a 2% increase of photon output from a 10 cm x 10 cm field to a 40 cm x 40 cm field. The off-axis location of the jaw opening does not have a significant effect on the magnitude of the backscatter. The backscatter effect is significant for monitor chambers using kapton windows, particularly for treatment fields using moving jaws. Applying the backscatter correction improves the accuracy of monitor-unit calculation using a model-based dose calculation algorithm such as the convolution method. PMID- 10798697 TI - Thermal and SAR characterization of multielement dual concentric conductor microwave applicators for hyperthermia, a theoretical investigation. AB - Six aperture array dual concentric conductor (DCO) microwave hyperthermia applicators were studied using theoretical models to characterize power deposition (SAR) and steady state temperature distributions in perfused tissue. SAR patterns were calculated using the finite difference time domain (FDTD) numerical method, and were used as input to a finite difference thermal modeling program based on the Pennes Bio-Heat Equation in order to calculate corresponding temperature distributions. Numerous array configurations were investigated including the use of different size DCC apertures (2, 3, and 4 cm), different spacing between apertures (1.0-2.0 cm), and different water bolus thicknesses (5 15 mm). Thermal simulations were repeated using blood perfusion values ranging from 0.5 to 5 kg/m3 s. Results demonstrate the ability of DCC array applicators to effectively and uniformly heat tissue down to a depth of 7.5-10 mm below the skin surface for a large number of different combinations of DCC element size, spacing, and water bolus thickness. Results also reveal the close correlation between SAR patterns and corresponding temperature distributions, verifying that design studies of the applicator can be performed confidently by analysis of SAR, from which the thermal behavior can be estimated. These simulations are useful in the design optimization of large microwave DCC array applicators for superficial tissue heating and for identifying appropriate aperture spacing and bolus thickness parameters for different size DCC aperture arrays and tissue blood perfusion conditions. PMID- 10798698 TI - Advanced single-slice rebinning in cone-beam spiral CT. AB - To achieve higher volume coverage at improved z-resolution in computed tomography (CT), systems with a large number of detector rows are demanded. However, handling an increased number of detector rows, as compared to today's four-slice scanners, requires to accounting for the cone geometry of the beams. Many so called cone-beam reconstruction algorithms have been proposed during the last decade. None met all the requirements of the medical spiral cone-beam CT in regard to the need for high image quality, low patient dose and low reconstruction times. We therefore propose an approximate cone-beam algorithm which uses virtual reconstruction planes tilted to optimally fit 180 degrees spiral segments, i.e., the advanced single-slice rebinning (ASSR) algorithm. Our algorithm is a modification of the single-slice rebinning algorithm proposed by Noo et al. [Phys. Med. Biol. 44, 561-570 (1999)] since we use tilted reconstruction slices instead of transaxial slices to approximate the spiral path. Theoretical considerations as well as the reconstruction of simulated phantom data in comparison to the gold standard 180 degrees LI (single-slice spiral CT) were carried out. Image artifacts, z-resolution as well as noise levels were evaluated for all simulated scanners. Even for a high number of detector rows the artifact level in the reconstructed images remains comparable to that of 180 degrees LI. Multiplanar reformations of the Defrise phantom show none of the typical cone-beam artifacts usually appearing when going to larger cone angles. Image noise as well as the shape of the respective slice sensitivity profiles are equivalent to the single-slice spiral reconstruction, z-resolution is slightly decreased. The ASSR has the potential to become a practical tool for medical spiral cone-beam CT. Its computational complexity lies in the order of standard single-slice CT and it allows to use available 2D backprojection hardware. PMID- 10798699 TI - A method for including redundant data in computed tomography. AB - A method for including redundant data in fan-beam computed tomography (CT) is presented. It is a natural extension of the Parker [Med. Phys. 9, 254-257 (1982)] short-scan approach applied to divergent fan-beam (or cone-beam) data when the data set covers between the minimally complete set of 180 degrees plus fan angle and 360 degrees. A virtual fan angle is introduced whose value is the difference between the angular range of the data collected and 180 degrees. Parker-weights are then applied as if the field-of-view is spanned by the virtual fan angle. PMID- 10798700 TI - Localization of cochlear implant electrodes in radiographs. AB - Multielectrode cochlear implantation is the most effective treatment for profound sensorineural hearing loss. In vivo three-dimensional 3-D localization of cochlear implant electrodes is important for modeling of the electrical field in the cochlea, design of electrode arrays, and may improve speech processor programming for better speech recognition. The prerequisite for 3-D localization of the electrodes is their 2-D localization in x-ray radiographs. In this paper, we develop a practical method to localize the electrodes with high efficiency, accuracy, and reproducibility. In this method, a priori knowledge of the electrodes and their approximate positions are utilized, an intelligent thresholding and segmentation mechanism is embedded, and the electrode center is computed as the weighted geometric center of segmented electrode pixels. Experiments with physical phantoms and human data demonstrate the feasibility and utility of this method. The PC-based program developed for this project is disseminated on the Web. PMID- 10798702 TI - Intraoperative ultrasound for guidance and tissue shift correction in image guided neurosurgery. AB - We present a surgical guidance system that incorporates pre-operative image information (e.g., MRI) with intraoperative ultrasound (US) imaging to detect and correct for brain tissue deformation during image-guided neurosurgery (IGNS). Many interactive IGNS implementations employ pre-operative images as a guide to the surgeons throughout the procedure. However, when a craniotomy is involved, tissue movement during a procedure can be a significant source of error in these systems. By incorporating intraoperative US imaging, the target volume can be scanned at any time, and two-dimensional US images may be compared directly to the corresponding slice from the pre-operative image. Homologous points may be mapped from the intraoperative to the pre-operative image space with an accuracy of better than 2 mm, enabling the surgeon to use this information to assess the accuracy of the guidance system along with the progress of the procedure (e.g., extent of lesion removal) at any time during the operation. Anatomical features may be identified on both the pre-operative and intraoperative images and used to generate a deformation map, which can be used to warp the pre-operative image to match the intraoperative US image. System validation is achieved using a deformable multi-modality imaging phantom, and preliminary clinical results are presented. PMID- 10798701 TI - In vivo biodistribution of 125IPIP and internal dosimetry of 123IPIP radioiodinated agents selective to the muscarinic acetylcholinergic receptor complex. AB - The development of new radioiodinated ligands for imaging the muscarinic acetylcholinergic complex (mAChR) using single photon emission computed tomography (SPECT) requires the evaluation of human organ doses prior to approval for human use. Animal biodistribution and excretion data were obtained and evaluated for IPIP, a new mAChR agent. Preliminary biodistribution studies were performed on four different stereoisomers of IPIP. A biokinetic model of the Z (S)-IPIP stereoisomer was constructed for the rat and used to estimate the internal absorbed dose in humans based on an extrapolation of the rat model. The thyroid is the critical organ for this radiopharmaceutical, with an absorbed dose estimate of 2.4 mGy/MBq for both males and females, when labeled with 123I. Even when blocked, the thyroid is still the critical organ, yet with a 90% dose reduction. The heart and brain receive the next highest doses in both males and females. Effective dose estimates for the use of pure 123I-PIP in humans are 0.16 mSv/MBq for males and 0.14 mSv/MBq for females. The biodistribution studies of the Z-(S)-IPIP stereoisomer showed the most promise as a successful agent for imaging muscarinic receptor sites in the heart and brain. IPIP also demonstrated potential as a therapeutic radiopharmaceutical for some colon carcinomas where muscarinic receptor sites are expressed in the tumor cells. These results provide preliminary data for use of IPIP in clinical studies on humans. PMID- 10798703 TI - Near-infrared study of fluctuations in cerebral hemodynamics during rest and motor stimulation: temporal analysis and spatial mapping. AB - We have noninvasively studied the motor cortex hemodynamics in human subjects under rest and motor stimulation conditions using a multichannel near-infrared tissue spectrometer. Our instrument measures optical maps of the cerebral cortex at two wavelengths (758 and 830 nm), with an acquisition time of 160 ms per map. We obtained optical maps of oxy- and deoxy-hemoglobin concentration changes in terms of amplitudes of folding average, power spectrum and coherence at the stimulation repetition frequency, and the phase synchronization index. Under periodic motor stimulation conditions, we observed coherence and frequency or phase synchronization of the local hemodynamic changes with stimulation. Our main findings are the following: (1) The amplitude of the hemodynamic response to the motor stimulation is comparable to the amplitude of the fluctuations at rest. (2) The spatial patterns of the oxy- and deoxy-hemoglobin responses to the stimulation are different. (3) The hemodynamic response to stimulation shows a spatial localization and a level of phase synchronization with the motor stimulation that depends on the stimulation period. PMID- 10798704 TI - Addendum to "Relevance of accurate Monte Carlo modeling in nuclear medical imaging" [Med. Phys. 26, 574-608 (1999)]. PMID- 10798705 TI - Comment on "Subunity coordinate translation with Fourier transform to achieve efficient and quality three-dimensional medical image interpolation" [Med. Phys. 26, 1776-1782 (1999)]. PMID- 10798706 TI - A fast fixed-point algorithm for independent component analysis of complex valued signals. AB - Separation of complex valued signals is a frequently arising problem in signal processing. For example, separation of convolutively mixed source signals involves computations on complex valued signals. In this article, it is assumed that the original, complex valued source signals are mutually statistically independent, and the problem is solved by the independent component analysis (ICA) model. ICA is a statistical method for transforming an observed multidimensional random vector into components that are mutually as independent as possible. In this article, a fast fixed-point type algorithm that is capable of separating complex valued, linearly mixed source signals is presented and its computational efficiency is shown by simulations. Also, the local consistency of the estimator given by the algorithm is proved. PMID- 10798707 TI - A study of early stopping and model selection applied to the papermaking industry. AB - This paper addresses the issues of neural network model development and maintenance in the context of a complex task taken from the papermaking industry. In particular, it describes a comparison study of early stopping techniques and model selection, both to optimise neural network models for generalisation performance. The results presented here show that early stopping via use of a Bayesian model evidence measure is a viable way of optimising performance while also making maximum use of all the data. In addition, they show that ten-fold cross-validation performs well as a model selector and as an estimator of prediction accuracy. These results are important in that they show how neural network models may be optimally trained and selected for highly complex industrial tasks where the data are noisy and limited in number. PMID- 10798708 TI - Neural networks in analog hardware--design and implementation issues. AB - This paper presents a brief review of some analog hardware implementations of neural networks. Several criteria for the classification of general neural networks implementations are discussed and a taxonomy induced by these criteria is presented. The paper also discusses some characteristics of analog implementations as well as some trade-offs and issues identified in the work reviewed. Parameters such as precision, chip area, power consumption, speed and noise susceptibility are discussed in the context of neural implementations. A unified review of various "VLSI friendly" algorithms is also presented. The paper concludes with some conclusions drawn from the analysis of the implementations presented. PMID- 10798709 TI - Inductive learning approaches to rainfall-runoff modelling. AB - Trying to model the rainfall-runoff process is a complex activity as it is influenced by a number of implicit and explicit factors--for example, precipitation distribution, evaporation, transpiration, abstraction, watershed topography, and soil types. However, this kind of forecasting is particularly important as it is used to predict serious flooding, estimate erosion and identify problems associated with low flow. Inductive learning approaches (e.g. decision trees and artificial neural networks) are particularly well suited to problems of this nature as they can often interpret underlying factors (such as seasonal variations) which cannot be modelled by other techniques. In addition, these approaches can easily be trained on the explicit factors (e.g. rainfall) and the inexplicit factors (e.g. abstraction) that affect river flow. Inductive learning approaches can also be extended to account for new factors that emerge over a period of time. This paper evaluates the application of decision trees and two artificial neural network models (the multilayer perceptron and the radial basis function network) to river flow forecasting in two flood prone UK catchments using real hydrometric data. Comparisons are made between the performance of these approaches and conventional flood forecasting systems. PMID- 10798710 TI - Curved trajectory prediction using a self-organizing neural network. AB - Existing neural network models are capable of tracking linear trajectories of moving visual objects. This paper describes an additional neural mechanism, disfacilitation, that enhances the ability of a visual system to track curved trajectories. The added mechanism combines information about an object's trajectory with information about changes in the object's trajectory, to improve the estimates for the object's next probable location. Computational simulations are presented that show how the neural mechanism can learn to track the speed of objects and how the network operates to predict the trajectories of accelerating and decelerating objects. PMID- 10798711 TI - Autonomous learning based on cost assumptions: theoretical studies and experiments in robot control. AB - Autonomous learning techniques are based on experience acquisition. In most realistic applications, experience is time-consuming: it implies sensor reading, actuator control and algorithmic update, constrained by the learning system dynamics. The information crudeness upon which classical learning algorithms operate make such problems too difficult and unrealistic. Nonetheless, additional information for facilitating the learning process ideally should be embedded in such a way that the structural, well-studied characteristics of these fundamental algorithms are maintained. We investigate in this article a more general formulation of the Q-learning method that allows for a spreading of information derived from single updates towards a neighbourhood of the instantly visited state and converges to optimality. We show how this new formulation can be used as a mechanism to safely embed prior knowledge about the structure of the state space, and demonstrate it in a modified implementation of a reinforcement learning algorithm in a real robot navigation task. PMID- 10798712 TI - Copper-dependent DNA damage induced by hydrazobenzene, an azobenzene metabolite. AB - Hydrazobenzene is carcinogenic to rats and mice and azobenzene is carcinogenic to rats. Hydrazobenzene is a metabolic intermediate of azobenzene. To clarify the mechanism of carcinogenesis by azobenzene and hydrazobenzene, we investigated DNA damage induced by hydrazobenzene, using 32P-5'-end-labeled DNA fragments obtained from the c-Ha-ras-1 protooncogene and the p53 tumor suppressor gene. Hydrazobenzene caused DNA damage in the presence of Cu(II). Piperidine treatment enhanced the DNA damage greatly, suggesting that hydrazobenzene caused base modification and liberation. However, azobenzene did not cause DNA damage even in the presence of Cu(II). Hydrazobenzene plus Cu(II) caused DNA damage frequently at thymine residues. Catalase and a Cu(I)-specific chelator inhibited Cu(II) mediated DNA damage by hydrazobenzene. Typical *OH scavengers did not inhibit the DNA damage. The main active species is probably a metal oxygen complex, such as Cu(I)-OOH. Formation of 8-oxo-7, 8-dihydro-2'-deoxyguanosine was increased by hydrazobenzene in the presence of Cu(II). Oxygen consumption and UV-Visible spectroscopic measurements have shown that hydrazobenzene is autoxidized to azobenzene with H2O2 formation. It is considered that the metal-mediated DNA damage by hydrazobenzene through H2O2 generation may be relevant for the expression of carcinogenicity of azobenzene and hydrazobenzene. PMID- 10798713 TI - Evidence for oxidised low density lipoprotein in synovial fluid from rheumatoid arthritis patients. AB - The oxidative modification of human LDL has been implicated in atherosclerosis, but the mechanisms by which such modification occurs in vivo are not fully understood. In the present study, we have isolated LDL from knee-joint synovial fluid of patients with rheumatoid arthritis. We demonstrate that such LDL is oxidatively modified as evidenced by an increased negative charge, distorted particulate nature and more rapid degradation by cultured macrophages. These results indicate that formation of oxidised LDL is associated with the local inflammatory response. Because the cellular interactions in rheumatoid arthritis have analogies with those in atherogenesis, we suggest that the rheumatoid joint is a useful model of atherosclerosis in which the in vivo process of LDL oxidation may be readily studied. PMID- 10798714 TI - The role of protein kinase C in the increased generation in isolated rat hepatocytes of the hydroxyl radical by puromycin aminonucleoside. AB - Puromycin aminonucleoside (PAN) has been known to induce proteinuria. The increased generation of reactive oxygen species (ROS) has been implicated in this toxicity of PAN. We have reported that PAN increases the synthesis of methylguanidine (MG) and creatol which are the products of the reaction of creatinine and the hydroxyl radical in isolated rat hepatocytes. However, the mechanism for the increased ROS induced by PAN is still unclear. In this paper, we investigate the role of protein kinase C (PKC) on the PAN induced reactive oxygen generation in isolated rat hepatocytes. Isolated hepatocytes were incubated in Krebs-Henseleit bicarbonate buffer containing 3% BSA, 16.6 mM creatinine and tested reagents. MG and creatol were determined by high performance liquid chromatography using 9,10-phenanthrenequinone for the post labeling. PAN increased MG and creatol synthesis in isolated rat hepatocytes by 60%. 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7), a PKC inhibitor, at 10 and 100 microM significantly inhibited MG and creatol synthesis with or without PAN. The inhibition rate is dose dependent from 10 to 100 microM. H1004, a reagent used as control for H-7, did not affect (at 10 microM) or increased little (at 100 microM) the synthesis of MG and creatol. Ro31-8425, a potent PKC inhibitor, significantly inhibited (at 10 microM) MG synthesis in the presence of PAN. PKC in the membrane fraction, a marker of PKC activation, increased over the initial concentration by a factor of 1.65-fold at 60 min incubation and 2.16-fold at 120 min with PAN, while it changed little without PAN. These results indicate that PAN activates PKC resulting in increased hydroxyl radical generation in isolated rat hepatocytes. PMID- 10798715 TI - Antioxidants and gadolinium chloride attenuate hepatic parenchymal and endothelial cell injury induced by low flow ischemia and reperfusion in perfused rat livers. AB - The objective of this study was to determine whether Kupffer cells contribute to parenchymal and endothelial cell damage induced by ischemia-reperfusion in perfused rat livers. Parenchymal and endothelial cell injury were determined by measuring activities of lactate dehydrogenase (LDH) and purine nucleoside phosphorylase (PNP), respectively, in the effluent perfusate of livers subjected to 60 min of low flow ischemia followed by 30 min of reperfusion. Upon reperfusion, LDH and PNP activities increased significantly within the first 10 min of reperfusion and remained elevated over control values throughout the duration of reperfusion. Pretreatment with gadolinium chloride, an inhibitor of Kupffer cell function, significantly decreased LDH and PNP efflux during reperfusion by approximately 60% and 50%, respectively. When Kupffer cells were stimulated by vitamin A pretreatment, PNP efflux was doubled during reperfusion. Vitamin E pretreatment attenuated LDH and PNP release by approximately 70% during reperfusion compared to enzyme release in untreated livers. Moreover, the water soluble antioxidants superoxide dismutase and desferrioxamine reduced reperfusion injury, whereas catalase had no effect on enzyme release. These results demonstrate that superoxide anions released from Kupffer cells are involved in oxidative damage to endothelial cells as well as hepatocytes during the early stages of hepatic reperfusion. PMID- 10798716 TI - Caspase activation and cytochrome c release during HL-60 cell apoptosis induced by a nitric oxide donor. AB - Nitric oxide (NO) from (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1- ium-1,2-diolate (NOC-18) induces apoptosis in human leukemia HL-60 cells. This effect was prevented by the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD-FMK), thereby implicating caspase activity in the process. NOC-18 treatment resulted in the activation of several caspases including caspase-3, -6, -8, and -9(-like) activities and the degradation of several caspase substrates such as nuclear lamins and SP120 (hnRNP-U/SAF-A). Moreover, release of cytochrome c from mitochondria was also observed during NOC 18-induced apoptosis. This change was substantially prevented by Z-VAD-FMK, thereby suggesting that the released cytochrome c might function not only as an initiator but also as an amplifier of the caspase cascade. Bid, a death agonist member of the Bcl-2 family, was processed by caspases following exposure of cells to NOC-18, supporting the above notion. Thus, NO-mediated apoptosis in HL-60 cells involves a caspase/cytochrome c-dependent mechanism. PMID- 10798717 TI - 4-Mercaptoimidazoles derived from the naturally occurring antioxidant ovothiols 1. Antioxidant properties. AB - 4-Mercaptoimidazoles derived from the naturally occurring family of antioxidants, the ovothiols, were assayed for their antioxidant properties. These compounds are powerful HOCl scavengers, more potent than the aliphatic thiol N-acetylcysteine. They react slowly with hydrogen peroxide with second order rate constants of 0.13 0.89 M(-1)s(-1). Scavenging of hydroxyl radical occurs at a diffusion-controlled rate (k=2.0-5.0 x 10(10)M(-1)s(-1)) for the most active compounds, which are also able to inhibit copper-induced LDL peroxidation. The combination of radical scavenging and copper chelating properties may explain the inhibitory effects on LDL peroxidation. Two molecules of mercaptoimidazole can chelate a copper ion and form a square planar complex detected by EPR. Compounds bearing an electron withdrawing group on position 2 of the imidazole ring are the most potent antioxidant molecules in this series. PMID- 10798718 TI - 4-Mercaptoimidazoles derived from the naturally occurring antioxidant ovothiols 2. Computational and experimental approach of the radical scavenging mechanism. AB - The radical-scavenging mechanism of fourteen 4-mercaptoimidazoles, derived from the natural family of ovothiols, was studied via a QSAR approach, cyclic voltammetry, ESR and NMR spectroscopy. A significant correlation was found between the DPPH scavenging abilities of test compounds and thermodynamic parameters like overall ease of disulphide formation. The production of a disulphide compound via thiyl radical formation is proposed. Upon DPPH scavenging, hydrogen abstraction from thiols yields transient short-lived thiyl radicals, which were characterised by ESR and rapidly dimerise to form a disulphide compound. Cyclic voltammetry showed that the best DPPH scavengers exhibit low oxidation potentials for their oxidation to disulphides. PMID- 10798719 TI - Application of confocal laser scanning microscopy to detect oxidative stress in human colon cells. AB - INTRODUCTION: Excess of intracellular reactive oxygen species in relation to antioxidative systems results in an oxidative environment which may modulate gene expression or damage cellular molecules. These events are expected to greatly contribute to processes of carcinogenesis. Only few studies are available on the oxidative/reductive conditions in the colon, an important tumour target tissue. It was the objective of this work to further develop methods to assess intracellular oxidative stress within human colon cells as a tool to study such associations in nutritional toxicology. METHODS: We have measured H2O2-induced oxidative stress in different colon cell lines, in freshly isolated human colon crypts, and, for comparative purposes, in NIH3T3 mouse embryo fibroblasts. Detection was performed by loading the cells with the fluorigenic peroxide sensitive dye 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate (diacetoxymethyl ester), followed by in vitro treatment with H2O2 and fluorescence detection with confocal laser scanning microscopy (CLSM). Using the microgel electrophoresis ("Comet") Assay, we also examined HT29 stem and clone 19A cells and freshly isolated primary colon cells for their relative sensitivity toward H2O2-induced DNA damage and for steady-state levels of endogenous oxidative DNA damage. RESULTS: A dose-response relationship was found for the H2O2-induced dye decomposition in NIH3T3 cells (7.8-125 microM H2O2) whereas no effect occurred in the human colon tumour cell lines HT29 stem and HT29 clone 19A (62-1000 M H2O2). Fluorescence was significantly increased at 62microM H2O2 in the human colon adenocarcinoma cell line Caco-2. In isolated human colon crypts, the lower crypt cells (targets of colon cancer) were more sensitive towards H2O2 than the more differentiated upper crypt cells. In contrast to the CLSM results, oxidative DNA damage was detected in both cell lines using the Comet Assay. Endogenous oxidative DNA damage was highest in HT29 clone 19A, followed by the primary colon cells and HT29 stem cells. CONCLUSIONS: Oxidative stress in colon cells leads to damage of macromolecules which is sensitively detected in the Comet Assay. The lacking response of the CLSM-approach in colon tumour cells is probably due to intrinsic modes of protective activities of these cells. In general, however, the CLSM method is a sensitive technique to detect very low concentrations of H2O2-induced oxidative stress in NIH3T3 cells. Moreover, by using colon crypts it provides the unique possibility of assessing cell specific levels of oxidative stress in explanted human tissues. Our results demonstrate that the actual target cells of colon cancer induction are indeed susceptible to the oxidative activity of H2O2. PMID- 10798720 TI - Comparison of the phenolic composition of fruit juices by single step gradient HPLC analysis of multiple components versus multiple chromatographic runs optimised for individual families. AB - After minimal sample preparation, two different HPLC methodologies, one based on a single gradient reversed-phase HPLC step, the other on multiple HPLC runs each optimised for specific components, were used to investigate the composition of flavonoids and phenolic acids in apple and tomato juices. The principal components in apple juice were identified as chlorogenic acid, phloridzin, caffeic acid and p-coumaric acid. Tomato juice was found to contain chlorogenic acid, caffeic acid, p-coumaric acid, naringenin and rutin. The quantitative estimates of the levels of these compounds, obtained with the two HPLC procedures, were very similar, demonstrating that either method can be used to analyse accurately the phenolic components of apple and tomato juices. Chlorogenic acid in tomato juice was the only component not fully resolved in the single run study and the multiple run analysis prior to enzyme treatment. The single run system of analysis is recommended for the initial investigation of plant phenolics and the multiple run approach for analyses where chromatographic resolution requires improvement. PMID- 10798721 TI - Comparison of identified leg motoneuron structure and function between larval and adult Manduca sexta. AB - Persistent leg motoneurons of the moth Manduca sexta were investigated in larval and adult animals to compare their dendritic structures, intrinsic electrical properties and pattern of target innervation. The study focused on two identified motoneurons of the prothoracic leg. Despite the complete remodeling of leg muscles, the motoneurons innervated pretarsal flexor muscles in both larval and adult legs. Similarly, although the central dendrites regress and regrow, the branching pattern was similar with the exception of a prominent midline branch that was not present in the adult stage. The intrinsic electrical properties of the motoneurons differed between larval and adult stages. Larval motoneurons had significantly higher membrane input resistances and more depolarized resting membrane potentials than did motoneurons in pharate adults or adults. In all stages, one motoneuron had a low maximal firing frequency, whereas the second motoneuron, which innervated the other half of the muscle, had a high maximum firing frequency. Although the two motoneurons continued to innervate the same halves of the target muscle, their relative effects on muscular contraction were reversed during metamorphosis along with concomitant changes in intrinsic properties. Pretarsal flexor motoneurons in pharate adults (just prior to emergence) displayed properties similar to those in emerged adults. PMID- 10798722 TI - The need for speed. I. Fast reactions and myelinated axons in copepods. AB - A rapid and powerful escape response decreases predation risk in planktonic copepods. Calanoid copepods are sensitive to small and brief hydrodynamic disturbances: they respond with multiple nerve impulses to a vibrating sphere. Some species, such as Pleuromamma xiphias and Labidocera madurae, respond with very large spikes (1-4 mV), whereas maximum spike heights are an order of magnitude smaller in others, such as Undinula vulgaris and Neocalanus gracilis. A comparative study of the escape responses showed that all species reacted within 10 ms of the initiation of a hydrodynamic stimulus. However, U. vulgaris and N. gracilis had significantly shorter reaction times (minimum reaction times: 1.5 ms and 1.6 ms) than the other two, P. xiphias (6.6 ms) and L. madurae (3.1 ms). Examination of the first antenna and the central nervous system using transmission electron microscopy revealed extensive myelination of sensory and motor axons in the two species with the shorter reaction times. Axons of the other two species resembled typical crustacean unmyelinated fibers. A survey of 20 calanoids revealed that none of the species in two of the more ancient superfamilies possessed myelin, but myelination was present in the species from three more recently-evolved superfamilies. PMID- 10798723 TI - The need for speed. II. Myelin in calanoid copepods. AB - Speed of nerve impulse conduction is greatly increased by myelin, a multi-layered membranous sheath surrounding axons. Myelinated axons are ubiquitous among the vertebrates, but relatively rare among invertebrates. Electron microscopy of calanoid copepods using rapid cryofixation techniques revealed the widespread presence of myelinated axons. Myelin sheaths of up to 60 layers were found around both sensory and motor axons of the first antenna and interneurons of the ventral nerve cord. Except at nodes, individual lamellae appeared to be continuous and circular, without seams, as opposed to the spiral structure of vertebrate and annelid myelin. The highly organized myelin was characterized by the complete exclusion of cytoplasm from the intracellular spaces of the cell generating it. In regions of compaction, extracytoplasmic space was also eliminated. Focal or fenestration nodes, rather than circumferential ones, were locally common. Myelin lamellae terminated in stepwise fashion at these nodes, appearing to fuse with the axolemma or adjacent myelin lamellae. As with vertebrate myelin, copepod sheaths are designed to minimize both resistive and capacitive current flow through the internodal membrane, greatly speeding nerve impulse conduction. Copepod myelin differs from that of any other group described, while sharing features of every group. PMID- 10798724 TI - Encoding of forces by cockroach tibial campaniform sensilla: implications in dynamic control of posture and locomotion. AB - Forces exerted by a leg in support and propulsion can vary considerably when animals stand upon or traverse irregular terrains. We characterized the responses of the cockroach tibial campaniform sensilla, mechanoreceptors which encode force via strains produced in the exoskeleton, by applying forces to the leg at controlled magnitudes and rates. We also examined how sensory responses are altered in the presence of different levels of static load. All receptors exhibit phasico-tonic discharges that reflect the level and rate of force application. Our studies show that: (1) tonic discharges of sensilla can signal the level of force, but accurate encoding of static loads may be affected by substantial receptor adaptation and hysteresis; (2) the absolute tonic sensitivities of receptors decrease when incremental forces are applied at different initial load levels; (3) phasic discharges of sensilla accurately encode the rate of force application; and (4) sensitivities to changing rates of force are strictly preserved in the presence of static loads. These findings imply that discharges of the sensilla are particularly tuned to the rate of change of force at all levels of leg loading. This information could be utilized to adapt posture and walking to varying terrains and unexpected perturbations. PMID- 10798725 TI - Visual pigments, oil droplets, ocular media and cone photoreceptor distribution in two species of passerine bird: the blue tit (Parus caeruleus L.) and the blackbird (Turdus merula L.). AB - The spectral absorption characteristics of the retinal photoreceptors of the blue tit (Parus caeruleus) and blackbird (Turdus merula) were investigated using microspectrophotometry. The retinae of both species contained rods, double cones and four spectrally distinct types of single cone. Whilst the visual pigments and cone oil droplets in the other receptor types are very similar in both species, the wavelength of maximum sensitivity (lambda(max)) of long-wavelength-sensitive single and double cone visual pigment occurs at a shorter wavelength (557 nm) in the blackbird than in the blue tit (563 nm). Oil droplets located in the long wavelength-sensitive-single cones of both species cut off wavelengths below 570 573 nm, theoretically shifting cone peak spectral sensitivity some 40 nm towards the long-wavelength end of the spectrum. This raises the possibility that the precise lambda(max) of the long-wavelength-sensitive visual pigment is optimised for the visual function of the double cones. The distribution of cone photoreceptors across the retina, determined using conventional light and fluorescence microscopy, also varies between the two species and may reflect differences in their visual ecology. PMID- 10798726 TI - Neural inhibition sharpens auditory spatial selectivity of bat inferior collicular neurons. AB - This study examines the role of neural inhibition in auditory spatial selectivity of inferior collicular neurons of the big brown bat, Eptesicus fuscus, using a two-tone inhibition paradigm. Two-tone inhibition decreases auditory spatial response areas but increases the slopes of directional sensitivity curves of inferior collicular neurons. Inferior collicular neurons have either directionally-selective or hemifield directional sensitivity curves. A directionally-selective curve always has a peak which is at least 50% larger than the minimum. A hemifield directional sensitivity curve rises from an ipsilateral angle by more than 50% and either reaches a plateau or declines by less than 50% over a range of contralateral angles. Two-tone inhibition does not change directionally-selective curves but changes most hemifield directional sensitivity curves into directionally-selective curves. Auditory spatial selectivity determined both with and without two-tone inhibition increases with increasing best-excitatory frequency. Sharpening of auditory spatial selectivity by two-tone inhibition is larger for neurons with smaller differences between excitatory and inhibitory best frequencies. The effect of two-tone inhibition on auditory spatial selectivity increases with increasing inhibitory tone intensity but decreases with increasing intertone interval. The implications of these findings in bat echolocation are discussed. PMID- 10798727 TI - Temperature and the temporal resolving power of fly photoreceptors. AB - A hot head gives an insect a clearer view of a moving world because warming reduces motion blur by accelerating photoreceptor responses. Over a natural temperature range, 19-34 degrees C, the speed of response of blowfly (Calliphora vicina) photoreceptors more than doubles, to produce the fastest functional responses recorded from an ocular photoreceptor. This acceleration increases temporal resolving power, as indicated by the corner frequency of the response power spectrum. When light adapted, the corner frequency increases from 53 Hz to 119 Hz with a Q10 of 1.9, and when dark adapted from 8 Hz to 32 Hz with a Q10 of 3.0. Temperature sensitivity originates in the phototransduction cascade, and is associated with signal amplification. The temperature sensitivity of photoreceptors must be taken into account when studying the mechanisms, function and ecology of vision, and gives a distinct advantage to insects that thermoregulate. PMID- 10798728 TI - Behavioral evidence of the role of lumbosacral anatomical specializations in pigeons in maintaining balance during terrestrial locomotion. AB - In birds there are anatomical specializations in the lumbosacral vertebrae and spinal cord. These include segmentally organized bony canals which are related to accessory lobes of the spinal cord. Both structures are connected by cerebrospinal fluid. To test whether these specializations function as a sense organ of equilibrium the effect of opening the fluid space was studied in pigeons. Locomotory behaviors on the ground (landing on a perch, keeping balance on a rotating perch, walking) but not flight were significantly impaired after lesion. These results support the assumption that the lumbosacral specializations are involved in the control of locomotion on the ground. PMID- 10798729 TI - Immunomodulation by fetal kidney cells: a reflection of immature antigen presenting cells? PMID- 10798730 TI - Unravelling the role of histocompatibility in liver transplantation. PMID- 10798731 TI - Regulation of xeno-reactive CD4+ T cells by CD8+CD28- T suppressor cells. PMID- 10798732 TI - Place of transplantation in infant intestinal failure management. PMID- 10798733 TI - To C or not to C: these are the questions in renal transplantation. PMID- 10798734 TI - Tumor necrosis factor polymorphisms and acute rejection: proven link or provisional association? PMID- 10798735 TI - Leukocyte activation and early dysfunction of liver grafts. PMID- 10798736 TI - A predictive parameter for good graft survival with limited relevance? PMID- 10798737 TI - Establishment of stable multilineage hematopoietic chimerism and donor-specific tolerance without irradiation. AB - BACKGROUND: Induction of tolerance to organ transplants will increase graft survival and decrease patient mortality and morbidity. Radiation-induced cytoreduction/ablation followed by donor hematopoietic cell reconstitution has been the most consistently successful approach to experimental tolerance induction. However, reluctance of clinicians to expose recipients to radiation has hampered its clinical application. METHODS: In the studies described, administration of polyclonal antilymphocyte serum (ALS), donor-specific bone marrow (DSBM) (150x10(6) cells), and sirolimus (24 mg/kg) in a completely mismatched murine model (B10.A donor, C57B/10 recipient) produced 100% indefinite (>250 days) skin graft survival. The level and character of donor-specific chimerism was evaluated with flow cytometry. RESULTS: Specific tolerance was confirmed by continued acceptance of primary and secondary donor-specific skin allografts and rejection of third-party grafts. The level and duration of chimerism induced was directly related to the dose of DSBM administered. Mice given 150x10(6) DSBM cells showed levels of 8-10% donor peripheral blood mononuclear cell chimerism by 30 days, and these levels persisted indefinitely (>250 days) in association with permanent tolerance of donor grafts. Eighty percent of donor chimeric cells were B lymphocytes (MHC class I and II positive, Fc receptor positive, CD45/B220 positive but negative for CD4, CD8 and Thy 1.2) and 20% were sorted in the macrophage monocyte population. CONCLUSIONS: These studies demonstrate for the first time that cytoreduction/ablation with ALS combined with sirolimus and reconstitution with donor bone marrow induces tolerance and chimerism in a completely mismatched murine combination. The use of ALS and sirolimus, currently employed therapies in clinical transplantation, and the lack of requirement for radiation make this tolerance protocol attractive for clinical application. PMID- 10798738 TI - Sirolimus in association with mycophenolate mofetil induction for the prevention of acute graft rejection in renal allograft recipients. AB - INTRODUCTION: A previous trial in renal transplantation comparing sirolimus (rapamycin) to cyclosporine (CsA) in a triple-drug therapy regimen with azathioprine and corticosteroids found that the incidence of acute rejection was similar (approximately 40%) with a trend for better renal function with sirolimus. METHODS: In 14 European centers, first cadaveric renal allograft recipients were randomized to receive sirolimus (n = 40) or CsA (n = 38) in an open-label design. All patients received corticosteroids and mycophenolate mofetil 2 g/day. Sirolimus and CsA were concentration controlled; trough levels of mycophenolic acid and prednisolone were also measured. RESULTS: At 12 months, graft survival (92.5% sirolimus vs. 89.5% CsA), patient survival (97.5% sirolimus vs. 94.7% CsA), and the incidence of biopsy-proven acute rejection (27.5% sirolimus vs. 18.4% CsA) were not statistically different. The use of antibodies to treat suspected rejection episodes was also similar (7.5% sirolimus vs. 5.3% CsA). More sirolimus patients received bolus steroid therapy (20 vs. 11, P = 0.068). From month 2 onward, the calculated glomerular filtration rate was consistently higher in sirolimus-treated patients. The adverse events reported more frequently with sirolimus were thrombocytopenia (45% vs. 8%) and diarrhea (38% vs. 11%). In the CsA group, increased creatinine (18% vs. 39%), hyperuricemia (3% vs. 18%), cytomegalovirus infection (5% vs. 21%), and tremor (5% vs. 21%) were observed significantly more often. DISCUSSION: Patient and graft survival and the incidence of biopsy-proven acute rejection at 12 months were comparable between sirolimus and CsA, whereas safety profiles were different. These data suggest that sirolimus may be used as primary therapy for the prevention of acute rejection. PMID- 10798739 TI - 24-hour lung preservation: simplified versus conventional University of Wisconsin solution in a porcine model. AB - BACKGROUND: Experimentally, the University of Wisconsin solution (UW) has been shown to be superior to the EuroCollins solution (EC) for lung graft preservation. We showed previously that the inclusion of the trisaccharide raffinose as an impermeant in the UW is largely responsible for this superiority. In this study, we used a new porcine model of isolated lung reperfusion to evaluate the use of a simple solution of phosphate-buffered raffinose (PBr) for lung preservation. METHODS: Lungs were stored for 24 hr at 4 degrees C after a single pulmonary artery flush with either UW (n = 5) or PBr (n = 5) solution. Left lungs were ventilated with room air and reperfused for 4 hr by venovenous extracorporeal circulation from a support animal. Controls (n = 5) were flushed with UW and reperfused without storage. RESULTS: Control lungs performed better than those stored in either solution in terms of oxygenation (P = 0.034) and airway pressure (P = 0.032). There were no significant differences between the two stored groups for any parameters. Data for stored lungs after 4 hr of reperfusion (means with 95% confidence intervals) include oxygenation (mm Hg): control 101.6 (14.5), UW 85.2 (14.5), PBr 75.0 (14.5); blood flow (ml/ min): control 572 (90), UW 466 (90), PBr 468 (90); peak airway pressure (mm Hg): control 15.9 (3.0), UW 21.0 (3.0), PBr 22.6 (3.0); pulmonary artery pressure (mm Hg): control 17.5 (3.2), UW 22.3 (2.9), PBr 24.5 (2.9). Graft edema (percentage tissue water): control 86.4 (0.8), UW 89.9 (1.8), PBr 89.3 (1.0). CONCLUSION: PBr is a far simpler and less expensive alternative to UW, and appears to provide a similar level of lung graft protection. PMID- 10798740 TI - Time-dependent impairment of mitochondrial function after storage and transplantation of rabbit kidneys. AB - BACKGROUND: The mitochondrial respiratory chain is implicated as a major target of kidney damage after ischemia-reperfusion. This study measures changes in integrated mitochondrial function and in the activity of enzymes of the respiratory chain after cold storage and transplantation-reperfusion in vivo. METHODS: Mitochondrial oxygen consumption and activities of respiratory chain enzymes and citrate synthase were measured in cortical mitochondria isolated from rabbit kidneys after 1-48 hr of cold ischemia with or without transplantation reperfusion. RESULTS: State 4 mitochondrial oxygen consumption was significantly increased after 48 hr of ischemia or 24-48 hr of ischemia with transplantation. Prolonged (24 or 48 hr) ischemic storage with and without transplantation caused a significant decrease in state 3 oxygen consumption, as did transplantation after 1, 24, and 48 hr of cold storage. Complex I and complex II-III activity decreased after 24 or 48 hr of ischemia, with transplantation having little additional effect. Complex IV activity was significantly decreased after 48 hr of ischemia, this decrease being exacerbated by transplantation-reperfusion. Complex V activity decreased significantly after 1 hr of ischemia and continued to decrease after 24-48 hr of ischemia. Transplantation after 1-24 hr (but not 48 hr) of ischemia resulted in partial recovery of complex V activity. Citrate synthase activity was decreased significantly only after 48 hr of ischemia and reperfusion, consistent with the loss of mitochondrial membrane integrity seen in electron micrographs of the transplanted 48-hr group. CONCLUSIONS: These data suggest that individual rabbit kidney mitochondrial complexes have different susceptibilities to cold ischemic and reperfusion damage. PMID- 10798741 TI - Effect of tacrolimus (FK506) and sirolimus (rapamycin) mono- and combination therapy in prolongation of renal allograft survival in the monkey. AB - BACKGROUND: Our previous studies confirmed that tacrolimus (FK506) and sirolimus [rapamycin (RAPA)], in combination, are not antagonistic but are synergistic in the prolongation of heart and small bowel grafts in the rodent. The aim of this study was to confirm further the synergistic effect of combined FK506 and RAPA in the more clinically relevant model, kidney transplantation in monkeys. METHODS: A total of 60 male Vervet monkeys were randomly assigned to 10 groups (n> or =5). Monkeys with renal allografts were treated with different doses of FK506 and/or RAPA orally for 60 days. Graft survival, body weight, clinical biochemistry determinations, oral glucose tolerance test, trough levels of the two drugs, and histopathology were investigated. RESULTS: Low doses of FK506 (1 or 4 mg/kg) combined with RAPA (0.5 mg/kg) produced synergistic effect in the prolongation of renal graft survival [combination index (CI) = 0.292, 0.565]. There were no additive or synergistic drug-associated toxicities such as hyperglycemia, nephrotoxicity, and hyperlipidemia. There also was no pharmacological antagonism. CONCLUSION: Concomitant therapy of low-dose (drug-optimal) FK506 and RAPA produced a synergistic effect in the prolongation of kidney allograft survival in Vervet monkeys without additive drug-associated toxicities. PMID- 10798742 TI - The protective role of Kupffer cells in humoral injury of xenoperfused rat livers. AB - BACKGROUND: The role of Kupffer cells in a hepatic xenograft rejection is still unclear. We investigated the effect of blocking Kupffer cells on xenogeneic humoral injury using rat livers as the xenoperfusion models. METHODS: Rat livers were perfused with fresh human blood after pretreatment either with normal saline (group 1; n = 8) or with gadolinium chloride (GdCl3) solution (group 2; n = 8). Tissue injury was evaluated by alanine aminotransferase release and histological examination. Tumor necrosis factor-alpha (TNF-alpha) production from rat livers was measured by enzyme-linked immunosorbent assay and also examined by immunohistochemistry. In addition, Kupffer cells were isolated after pretreatment either with normal saline or with GdCl3 solution and incubated with human serum. Localization of human C3 and IgM was examined by immunofluorescence. RESULTS: Alanine aminotransferase release in group 2 was significantly higher than in group 1 (P = 0.015). Histological examination revealed more severe tissue injury in group 2. The mean TNF-alpha level was not significantly different between the two groups. In immunohistochemistry, TNF-alpha was positive primarily on vascular endothelial cells in both groups. Immunofluorescence of saline-treated Kupffer cells showed an uptake of human C3 in the cytoplasm, whereas no uptake was observed in GdCl3-treated cells. The uptake of human IgM did not differ between the two groups. CONCLUSIONS: These results suggest that Kupffer cells have a protective role in preventing xenogeneic humoral injury. Their ability to absorb xenogeneic complements may contribute to this protective mechanism. PMID- 10798743 TI - Cyclosporine A and azathioprine are equipotent in chronic kidney allograft rejection. AB - Chronic rejection is the major cause of graft loss after kidney transplantation. Various immunosuppressive protocols have been used to ameliorate this process. We investigated whether cyclosporin A- (CyA) or azathioprine- (Aza) based immunosuppression is better able to slow the progression of chronic rejection. Fisher kidneys were transplanted into bilaterally nephrectomized Lewis rats. Recipients received CyA (1.5 mg/kg/day, s.c.) for 10 days, and were treated from day 11 with either CyA (1.5 mg/kg)+pred (0.15 mg/kg) (C+P),Aza (2 mg/kg)+pred (A+P), vehicle+pred (P), or vehicle alone (controls) (n = 8/group). Proteinuria was regularly assessed and grafts were harvested for morphological, immunohistological, and molecular biological analysis at week 24. By week 12 proteinuria had increased to significant levels. At week 24, proteinuria was significantly lower and creatinine clearance was significantly higher in C+P and A+P, than in P or controls. Morphological analysis supported these functional results: at week 24, glomerulopathy, tubular atrophy and intimal proliferation (as assessed according to the BANFF score) were less pronounced in C+P and A+P, as compared with P or controls. These morphological parameters were accompanied by a reduced infiltration of ED-1+ macrophages and CD-5+ T lymphocytes. In P or controls the synthesis of IL-2Ralpha mRNA was markedly elevated at this time. In parallel to the reduced cellular infiltration, IL-2Ralpha mRNA expression was markedly inhibited, both, in C+P and A+P. There were no significant differences between C+P and A+P regarding the parameters studied. In conclusion, both C+P and A+P reduced the infiltration of activated T lymphocytes, and the pace of chronic kidney allograft rejection. The outcome of C+P and A+P based therapy did not differ significantly. PMID- 10798744 TI - Tolerance induced by bone marrow chimerism prevents transplant vascular sclerosis in a rat model of small bowel transplant chronic rejection. AB - BACKGROUND: The major impediment to success in solid organ transplantation is chronic rejection (CR). The characteristic lesion of CR is transplant vascular sclerosis (TVS). Although the mechanism of TVS is thought to have an immunologic basis, in humans immunosuppression does not prevent or reverse it. One possible therapy to prevent TVS is induction of donor-specific tolerance. Bone marrow chimerism has been successful in inducing tolerance in acute and chronic rejection heart and kidney transplant models. The highly immunogenic small bowel (SB) allograft provides a rigorous test of the efficacy of this tolerance regimen. We examined whether induction of tolerance by bone marrow chimerism could prevent TVS in a model of Fisher 344 (F344) to Lewis (LEW) rat SB transplantation. METHODS: Bone marrow chimeras (BMC) were created by transplantation of T-cell-depleted F344 bone marrow into irradiated LEW rats. Chimerism was assessed by flow cytometric method. F344 SB, heterotopically transplanted into the chimeras, was clinically and histologically assessed for CR. F344 SB grafts, transplanted into cyclosporine-A-treated LEW recipients, served as control grafts for CR. RESULTS: Cyclosporine-A-treated LEW rats chronically rejected F344 SB grafts. By contrast, the BMC group demonstrated tolerance and had long-term SB graft survival (>120 days) without TVS. The BMC demonstrated immunocompetence by prompt rejection of third party ACI (RT1av1) SB allografts. CONCLUSIONS: Bone marrow chimerism prevents chronic graft failure secondary to TVS in a model of chronic SB rejection. TVS fails to develop when tolerance is established, suggesting that the mechanisms involved in TVS are, in part, immunologically mediated. PMID- 10798745 TI - Induction of xenoreactive CD4+ T-cell anergy by suppressor CD8+CD28- T cells. AB - BACKGROUND: The underlying mechanism of immune suppression mediated by regulatory T cells is not completely understood. In previous studies we have shown that antigen-specific human T suppressor cells (Ts) can be generated in vitro by multiple rounds of stimulation with allogeneic, xenogeneic, or antigen-pulsed autologous antigen-presenting cells (APC). Human Ts express the CD8+CD28- phenotype and require specific recognition of MHC class I/peptide complexes on the surface of APC to block proliferation of T helper cells (Th). The aim of the present study was to explore the activation requirements of Ts as well as the nature of Th unresponsiveness to xenogeneic (swine) antigens induced by Ts. METHODS AND RESULTS: We investigated whether specific antigenic stimulation of Ts is required for their ability to inhibit early activation of xenoreactive Th (up regulation of CD40 ligand). Flow cytometry studies indicated that Ts function required specific recognition of MHC class I on the surface of the stimulating APC. However, neither proliferation nor protein synthesis was required for the ability of Ts to inhibit Th. Ts drastically reduced the capacity of xenoreactive Th cells to produce interleukin (IL)-2 in response to the specific APC, without affecting their surface expression of IL-2 receptor. The suppressor effect that Ts exerted on Th proliferation could not be circumvented by CD40 ligation on the surface of the APC but could be reversed by the addition of exogenous IL-2. CONCLUSION: These data indicate that Ts induce anergy of xenoreactive human Th cells upon specific recognition of MHC class I antigens. Hence, Ts may prevent the activation of T cell-mediated immune responses against xenogeneic transplants. PMID- 10798747 TI - Insulin secretion and sensitivity after simultaneous pancreas-kidney transplantation estimated by continuous infusion of glucose with model assessment. AB - BACKGROUND: Monitoring of insulin secretion and sensitivity after pancreas transplantation remains a practical problem. METHODS: We introduced the simple structural model, continuous infusion of glucose with model assessment (CIGMA), to obtain insulin secretion and insulin sensitivity estimations after 35 successful simultaneous pancreas-kidney transplantations. Eighteen non-diabetic kidney transplant recipients were used as control group. RESULTS: The baseline characteristics were equal between the two groups except for higher fasting insulin levels in the pancreas transplant group. After the 1-hr CIGMA glucose load, the pancreas transplant group reached a mean +/- SD blood glucose of 8.2+/ 1.7 mmol/L compared with 7.3+/-1.0 mmol/L in the control group (P = 0.05). Concurrent stimulated insulin and C-peptide levels were 48+/-28 mU/L and 2.3+/ 0.9 nmol/L in the pancreas transplant group compared with 36+/-21 mU/L and 2.9+/ 1.1 nmol/L in the control group (P = 0.1 and P = 0.03, respectively). Both the CIGMA estimation for secretion as well as the CIGMA estimation for sensitivity were lower in pancreas transplant group (P = 0.003 and P = 0.01, respectively). Mean +/- SE coefficients of variation for the model estimations were 15+/-4% for secretion and 17+/-6% for sensitivity. CONCLUSIONS: We conclude that CIGMA can be used clinically to evaluate carbohydrate metabolism in pancreas-kidney transplant recipients. These patients have a reduction in insulin secretory capacity and evidence of more insulin resistance than non-diabetic kidney transplant recipients. PMID- 10798746 TI - Improving the preservation of isolated rat skeletal muscles stored for 16 hours at 4 degrees C. AB - BACKGROUND: Limiting factors for long-term cold preservation of isolated skeletal muscles are increased intracellular calcium levels, the occurrence of hypercontraction, and the overproduction of oxygen free radicals. In the present study, we investigated whether muscle preservation during cold storage could be improved by additives that can protect against such processes or by oxygen supply. METHODS: The soleus (SOL) and a strip of the cutaneus trunci muscle (CT) from the rat were isolated and stored for 16 hr at 4 degrees C in Bretschneider's Histidine Tryptophane Ketoglutarate (HTK) and subsequently acclimatized in Krebs Henseleit solution for 90 min at room temperature. The protective effects of 2,3 butanedione monoxime (BDM; reduces intracellular calcium release and inhibits fiber contraction) and of the following antioxidants were investigated: N-tert butyl-alpha-phenylnitrone (PBN), trolox, desferal, and deferione. The antioxidants and BDM were added to both HTK and Krebs-Henseleit solution. Dose response curves were made for each of the additives (n> or =4 for each dose). To evaluate the effect of oxygen supply, HTK was aerated with 95% O2/5% CO2. Muscle function (P0), energy metabolism (ATP), and cytoarchitecture were analyzed. The measured values were compared with those of fresh unstored muscles (% of control) and with those of muscles stored in HTK without any additive (multivariate analysis of variance, P<0.05). RESULTS: We found a significant protection of the contractile function (P0) of both muscles after the addition of 1 mM of trolox (SOL: 46% of control; CT: 53%) and after the addition of 3 mM or 0.3 mM of deferione to the SOL and CT, respectively (P0 for both muscles: 55%), whereas no protection was found with PBN (0.03-1 mM) and Desferal (0.001-1 mM). The addition of BDM (10 or 30 mM) resulted in the highest increase of P0 (84% and 60% for the SOL and CT, respectively). The combinations BDM-trolox and BDM-deferione did not further improve the preservation of the SOL function, but P0 values (88% and 91% of control, respectively) were not different from those found for control muscles. Oxygenation of HTK was only beneficial for the SOL (P0: 83%). The improved preservation of muscle function was accompanied by a reduction of the twitch threshold current, increased by storage, suggesting a protective effect of the intervention on the preservation of the muscle cell membrane integrity. Biochemical and histological data corresponded well with the functional data. CONCLUSIONS: The results showed that the addition of BDM and antioxidants (trolox and deferione) to the bathing solutions improved the preservation of the function, metabolism, and cytoarchitecture of isolated skeletal muscles after cold storage for 16 hr. PMID- 10798748 TI - Factors predicting the long-term success of maintenance cyclosporine monotherapy after kidney transplantation. AB - BACKGROUND: The theoretical aim of maintenance cyclosporine monotherapy (mCsA) after kidney transplantation is to reduce the incidence of the metabolic complications of corticosteroids and to minimize the adverse effects of excessive long-term immunosuppression. This study was performed in low-immunological-risk cadaveric kidney transplant recipients to evaluate the risks and benefits of mCsA and the long-term graft survival, and to determine the factors predicting success of this policy. METHODS: The multicenter retrospective study was conducted in 329 Caucasian patients receiving mCsA out of 728 first cadaveric kidney transplant recipients. The inclusion criteria were: HLA antibodies < or =25%, serum creatinine <200 micromol/L, and no rejection or only one rejection episode. At the end of the study, we compared the group of patients successfully treated with mCsA (successful group) with those requiring additional immunosuppressive agents (unsuccessful mCsA group). RESULTS: Overall patient and graft survival rates for the 728 first cadaveric graft were 92% and 64%, respectively, at 8 years. Out of 329 patients enrolled in mCsA, 240 were maintained on this treatment and 89 were withdrawn (3 deaths, 18 graft losses, 68 functional grafts). The 8-year graft survival in the 329 enrolled mCsA patients was 84%, 95% in the successful mCsA group, and 70% in the unsuccessful mCsA group. Multivariate analysis showed that the factors predicting success of mCsA were: donor age <40 years (P = 0.001), serum creatinine at mCsA initiation <125 micromol/L (P = 0.02), no rejection episode before mCsA initiation (P = 0.005), and glomerulopathy as the primary renal disease (P = 0.001). CONCLUSION: Numerous kidney transplant recipients with a low immunological risk and good and stable renal function may benefit from discontinuation of prednisone and azathioprine in order to reduce the complications related to these drugs. This therapeutic approach had no adverse impact on the overall long-term graft survival for "low risk" and successful patients. PMID- 10798749 TI - Impaired antigen-presenting cell function contributes to T-cell hyporesponsiveness in stable lung transplant recipients. AB - BACKGROUND: Peripheral blood mononuclear cells (PBMC) of stable renal or cardiac transplant recipients were previously shown to respond to allogeneic cells but not to soluble protein antigens. The aim of the present study was to assess the T cell and antigen-presenting cell (APC) functions of stable lung transplant (LT) recipients. METHODS: We obtained PBMC from 38 stable LT recipients. PBMC from healthy volunteers served as controls. PBMC were stimulated with either anti-CD3 monoclonal antibody, allogeneic PBMC, or tetanus toxoid (TT). T-cell activation was assessed by determination of interleukin (IL)-2 levels in culture supernatants; in some experiments, interferon-y levels were also determined. Patients' APC function was tested in a mixed leukocyte reaction using patients' PBMC as stimulators. The expression of class II MHC, B7.2, and CD40 molecules on patients' APC was determined by flow cytometry, and their production of IL-10 and IL-12 at the basal state and upon CD40 ligation was also measured. RESULTS: Patients' T cells produced normal amounts of IL-2 in response to anti-CD3 monoclonal antibody and allogeneic PBMC. In contrast, the response of memory T cells to TT was severely blunted both in terms of IL-2 and interferon-y production. Patients' PBMC were poor stimulators in mixed leukocyte reaction, and class II MHC expression on patients' monocytes was significantly reduced. Patients' APC presented a modest but significant increase in basal IL-10 production and produced significantly less IL-12 upon CD40 ligation than control APC. CONCLUSIONS: T cells from stable LT recipients respond normally to stimuli that do not depend on autologous APC. The major impairment in the T-cell response to TT is caused by APC dysfunction, which involves decreased class II MHC expression and deficient IL-12 synthesis. PMID- 10798750 TI - Graft-protective role of high pretransplantation IgA-anti-Fab autoantibodies: confirmatory evidence obtained in more than 4000 kidney transplants. The Collaborative Transplant Study. AB - BACKGROUND: Preliminary studies showed an excellent success rate of kidney grafts in patients with high pretransplantation serum levels of IgA autoantibodies directed against the Fab region of the human IgG molecule. METHODS: With the collaboration of 30 centers from around the world, we attempted to verify the role of IgA-anti-Fab autoantibodies in kidney transplantation in an entirely new series of 4316 cadaveric kidney transplants, with special consideration of presensitized and poorly HLA-matched recipients. RESULTS: In agreement with previously published preliminary findings, 147 recipients with a high pretransplantation IgA-anti-Fab of >1000 had a 2-year kidney graft survival rate of 88+/-3% (+/- SE), compared with an 80+/-1% rate in 851 recipients with a low IgA-anti-Fab of <60 (P = 0.02). Even in patients at an increased risk of graft rejection, high pretransplantation IgA-anti-Fab autoantibody activity was associated with superior graft survival. Among 815 presensitized patients, 31 had a high pretransplantation IgA-anti-Fab activity of >1000 and their 2-year graft survival rate was 94+/-4%, in contrast to a 71+/-4% rate in 165 presensitized recipients with a low IgA-anti-Fab of <60 (P = 0.02). Of 2294 recipients who received a kidney with > or =3 HLA-A+B+DR mismatches, 79 recipients had a high pretransplantation IgA-anti-Fab of >1000 and a 2-year graft survival rate of 90+/ 4%, as compared with a 79+/-2% rate in 459 patients with a low IgA-anti-Fab of <60 (P = 0.04). CONCLUSIONS: The present study confirms that kidney graft recipients with high pretransplantation IgA-anti-Fab activity have excellent graft survival, and it extends this observation to presensitized recipients and poor HLA matches. PMID- 10798751 TI - Immune reconstitution after allogeneic bone marrow transplantation depleted of T cells. AB - BACKGROUND: Immune reconstitution following transplantation in individuals who had received T-cell-depleted marrow from HLA identical siblings was serially documented and correlated with the clinical recovery. METHODS: Patients were preconditioned with radiation containing programs. GvHD prophylaxis was by T-cell depletion with CAMPATH 1G (ex vivo; median dose 20 mg). After transplantation lymphoid development was studied by flow cytometry and serum Ig concentrations were determined. Charts were reviewed to determine the effects of the immune reconstitution on the clinical performance. RESULTS: The mean donor mononuclear cell number infused was 0.89x10(8)/kg. Within 6 months all the patients recovered their blood parameters and only one required therapy for GvHD. However, despite normal blood counts, 15 suffered life-threatening opportunistic infections, developing at a median of 24 weeks post grafting, but occurring even after 11 months. At 8 weeks from marrow infusion when leukocyte values had normalized in 15/20, compared to normal, immunophenotyping of blood cells from BMT revealed a significantly reduced mean lymphocyte count (1.06, SD 0.83x10(9)/l; P = 0.01), cells expressing CD3 (0.7x10(9)/l, SD 0.68; P = 0.05), CD4 (0.13x10(9)/l, SD 0.21; P = 0.0001) and CD19 (0.04x10(9)/l, SD 0.05; P = 0.001). Populations expressing CD8 and CD56 remained within normal range throughout the study. Normalization of cell numbers displaying CD2, CD3 and CD19 was delayed until 52, 52 and 24 weeks respectively, while CD4 counts persisted subnormal even at 72 weeks. Serum IgA levels were significantly decreased for the entire study period. CONCLUSIONS: T-cell depletion with CAMPATH 1G while effectively preventing GvHD, also causes clinically significant and prolonged immunosuppression with apparently important clinical implications. PMID- 10798752 TI - Frequency, potential risk and therapeutic intervention in end-stage renal disease patients with antiphospholipid antibody syndrome: a multicenter study. AB - BACKGROUND: Antiphospholipid antibody syndrome (APAS) is characterized by the presence of anticardiolipin antibodies (ACA) in association with thrombotic disorders of arterial and/or venus systems, spontaneous abortion(s) or thrombocytopenia. METHODS: In this multicenter study, 502 end-stage renal disease (ESRD) patients awaiting renal transplants were screened to determine the frequency of APAS, the potential risk associated with APAS, and strategies for therapeutic intervention. Ninety-three patients (19%) had high titers of ACA. Twenty-three patients had documented evidence of one or more of the thrombotic disorders such as lupus, frequent abortions, frequent thrombosis of arteriovenous shunts, biopsy-proven microrenal angiopathy, or thrombocytopenia and thus were diagnosed with APAS. Of these 23 patients, 11 received kidney transplants either with (4 patients) or without (7 patients), concomitant anticoagulation therapy. RESULTS: All seven of the patients with APAS not treated with anticoagulation therapy lost their allografts within 1 week as a result of renal thrombosis. In contrast, three out of four transplant patients with APAS treated with anticoagulation therapy maintained their allografts for over 2 years. The fourth patient lost his graft within a week because of thrombosis. Of the remaining 70 patients with high titers of ACA but no evidence of thrombotic disorders, 37 received kidney transplants. None lost their allografts as a result of thrombosis. Our data suggest that, although 19% of our ESRD patients exhibit high titer of ACA, only 5% of the patients have APAS. CONCLUSION: In conclusion, our data suggest that the patients with APAS are at high risk of posttransplant renal thrombosis. Anticoagulation therapy could prevent patients from posttransplant thrombosis in patients with APAS. PMID- 10798753 TI - Biopsy of marginal donor kidneys: correlation of histologic findings with graft dysfunction. AB - BACKGROUND: Kidney biopsies are being used to evaluate marginal donors, but rigorous statistical validation of this practice with multivariate analysis has not been performed. METHODS: To analyze histologic parameters in 78 donor biopsies for their ability to predict graft dysfunction, we used a proportional odds model that included both donor and recipient factors. Glomerulosclerosis was categorized into grades 0, 1, 2, and 3, corresponding to 0, 1-10%, 11-20%, and 21 30% global sclerosis, respectively. The degrees of interstitial fibrosis, tubular atrophy, arteriosclerosis, and arteriolar hyalinosis were graded from 0 to 3+, using definitions suggested by the Banff Schema of allograft pathology. RESULTS: Increasing donor age was associated with higher glomerulosclerosis, tubular atrophy, and arteriosclerosis. Kidneys with any degree of interstitial fibrosis were 2.6 times [odds ratio (OR)] more likely to experience a worse outcome at 6 months (P = 0.02). This association held up after correction for acute rejection (OR 2.5, P = 0.03) and high panel-reactive antibody (OR 3.4, P = 0.006), However, the OR was reduced to 1.9 (P = 0.15) after controlling for recipient age. With each increment in the grade of glomerulosclerosis, the odds for a worse outcome at 12 months increased to 2.3 (P = 0.005). The value for OR became 2.0 (P = 0.03) when controlling for recipient age (P = 0.01), 2.4 (P = 0.005), when controlling for acute rejection, and 2.3 (P = 0.006) when controlling for high panel-reactive antibody. CONCLUSIONS: Histopathological parameters present in donor biopsies can independently predict post-transplant graft function. Implications for the pool of donor organs available for transplantation are discussed. PMID- 10798754 TI - Bone marrow transplantation depleted of T cells followed by repletion with incremental doses of donor lymphocytes for relapsing patients with chronic myeloid leukemia: a therapeutic strategy. AB - BACKGROUND: For patients with chronic myeloid leukemia (CML), long-term survival after stem cell transplantation requires adequate control of graft-versus-host disease (GVHD) and disease recurrence. Relapsing patients respond to donor lymphocyte infusion (DLI) but develop life-threatening complications. METHODS: Patients with CML in first chronic phase received bone marrow (n = 14) or peripheral blood progenitor cell transplants (n = 4) from HLA-identical siblings. GVHD prophylaxis was by ex vivo T-cell depletion with CAMPATH 1G. If disease recurred, donors' mononuclear cells were collected by apheresis, the CD3 samples commencing at 10(6)/kg were aliquoted at half-log increment intervals, cryopreserved, and infused until disease clearance. RESULTS: Eighteen patients (median age: 32.5 years) received transplants. All engrafted without procedure related mortality. Fourteen patients relapsed, and 13 entered the DLI program. Two developed extensive GVHD after single schedule infusions ranging from 89x10(6) to 670x10(6) mononuclear cells/kg, and one survives in complete remission (CR). The rest, treated with incremental dose DLI, experienced no acute toxicities. One, who had developed grade III steroid-responsive GVHD, died in CR2 from opportunistic infections. Steroids reversed limited cutaneous GVHD and elevated liver enzymes in five patients. Three others developed pancytopenia, and two restored blood counts only after donor peripheral blood progenitor cell infusions. Molecular CR2 was established in 12/13 patients, occurring in 10/11 (91%) on the incremental program at a median accumulation of 67 (range: 5-166) x10(6) CD3 cells/kg. Sixteen of 18 (89%) survive at median of 854.5 days from bone marrow transplantation, 4 in CR1 and 10 in CR2 at a median disease-free survival (for remission 2) duration of 341 days. The median combined disease-free survival of the 14 patients in CR 1+2 is 660 days, with 99% average performance status. CONCLUSIONS: Escalating DLI leads to safe new molecular CR in most CML relapse patients. These results raise the possibility of using "safe" transplantation programs of T-cell depletion, that include graded DLI as prevention against disease recurrence. PMID- 10798755 TI - A prospective randomized study of preoperative nutritional supplementation in patients awaiting elective orthotopic liver transplantation. AB - BACKGROUND: Poor nutritional status is common among patients awaiting orthotopic liver transplantation and is associated with poor outcome. METHODS: This prospective randomized controlled trial examined the effect of pretransplant nutritional supplementation on the outcome of patients undergoing liver transplantation. Of 82 consecutive patients with mid-arm muscle circumference <25th percentile, 42 received enteral supplementation, and the remainder acted as the control group. The supplemented group received a calorie-dense enteral feed taken daily (in addition to diet) until transplantation. Nutritional status was monitored by upper arm anthropometric measurements and handgrip strength. Dietary intake was calculated from 5-day food diaries. RESULTS: Supplementation improved mid-arm circumference, mid-arm muscle circumference, and grip strength. Pretransplant nutritional status was not associated with posttransplant sepsis or major complications. Preoperative grip strength of <85% of normal values was predictive of increased incidence of posttransplant major complications. Supplementation did not affect outcome, although there were more deaths in the control group (seven deaths before and two deaths after transplant) than there were in the supplemented group (two deaths before and three deaths after transplant). There was no difference in overall survival (P = 0.075). CONCLUSIONS: Enteral supplementation improved some parameters of nutritional status pretransplant. Dietary intake of patients in the two groups was similar at transplant. Nutritional supplementation has not increased nutritional intake, although this may reflect the importance of regular dietetic input and support, rather than suggesting that nutritional supplementation is ineffective. Supplementation had no effect on outcome of liver transplantation. PMID- 10798756 TI - Sensitization and sensitivity: defining the unsensitized patient. AB - BACKGROUND: Since the landmark studies of Patel and Terasaki in the late 1960s, pretransplant cross-matching has been performed by HLA laboratories on a 24-hr/7 day basis. In fact, regulating agencies such as the American Society for Histocompatibility and Immunogenetics and the United Network for Organ Sharing have mandated prospective crossmatching for selected solid organ transplants. However, two recent publications (Transplantation 1998; 66: 1833; and Transplantation 1998; 66: 1835) have suggested a change to this approach. Specifically, those authors advocate the transplantation of non-sensitized individuals without a final prospective cross-match as a means to reduce cold ischemia time and the incidence of delayed graft function. Such considerations were predicated upon results generated by cytotoxicity-based antibody screening. We and others, however, have reported that a flow cytometric-based assay is a more sensitive method to detect alloantibodies than cytotoxicity. Furthermore, an increasing number of reports document that graft survival is improved among patients whose final flow cytometric crossmatches were negative compared to patients with positive flow cytometric crossmatches. Although we agree that it is reasonable to transplant truly non-sensitized patients without a prospective final crossmatch, our data demonstrate that a large number of patients deemed non sensitized by cytotoxicity-based antibody assessment are, in fact, sensitized. METHODS: Panel-reactive antibody (PRA) testing was performed with 703 sera from 527 patients. The patient population consisted of individuals awaiting either renal or cardiac transplantation. PRA evaluations were performed using lymphocyte cytotoxicity (antiglobulin-enhanced, complement-dependent cytotoxicity [AHG-CDC]) or assays (enzyme-linked immunosorbent assay [ELISA]; flow cytometry) in which solubilized HLA molecules were affixed to solid phase matrices. RESULTS: PRA activity in 264 sera from 88 patients was evaluated by AHG-CDC, ELISA, and flow cytometry. Results among the three methods were concordant for 83% of these sera. Discordant results occurred with 32 samples and demonstrated a distinct hierarchy in the sensitivity of the three techniques to detect alloantibodies. None of the 32 sera were positive by AHG-CDC, 20/32 were positive by ELISA, and 32/32 were positive by flow cytometry. Subsequent studies revealed that, among 527 patients, 302 (57%) exhibited 0% PRA by AHG-CDC. Of these 302 AHG-CDC-negative patients, 76 (25%) had class I or class II antibodies detectable using a flow cytometric approach. Within the AHG-CDC-negative/flow cytometric-positive patients, PRA values exhibited a wide range (6-99%) for both class I and class II antibodies. The average PRA was 27% and 38% for class I and II, respectively. Retrospective flow cytometric crossmatches performed for 30 recipients of cardiac allografts whose AHG-CDC PRA were 0% revealed that 11/30 crossmatches were positive. CONCLUSIONS: The concept of transplanting non-sensitized patients without a prospective final crossmatch is appealing and, if bona fide, clearly makes sense. However, our data demonstrate that how a patient is deemed non-sensitized is critical. The difference between AHG- and flow cytometric-based PRA testing is significant and can result in transplantation of alloimmunized patients considered to be non-sensitized. Therefore, we recommend that, if a transplant center chooses to forego a prospective final crossmatch, the decision to do so should be based on methods more sensitive than AHG-CDC. PMID- 10798757 TI - Liver regeneration and function in donor and recipient after right lobe adult to adult living donor liver transplantation. AB - BACKGROUND: Regeneration of the liver to a predetermined size after resection or transplantation is a well described phenomenon, but the time course over which these events occur has not been well defined. It is not clear how initial liver mass, reperfusion, immunosuppression, or steatosis influence this process. METHODS: Liver regeneration was assessed prospectively by volumetric magnetic resonance imaging (MRI) in living right lobe liver donors and the recipients of these grafts. Imaging was performed at regular intervals through 60 days after resection/transplantation, and liver mass was determined. Liver function tests and synthetic function were monitored throughout the study period in donors and recipients of these grafts as well as recipients of cadaveric grafts. RESULTS: MRI consistently overestimated liver mass by a mean of 45 g (+/-65) (range 10 123). Donor liver mass increased by 101%, 110%, 115%, and 144% at 7, 14, 30, and 60 days after resection, respectively. Recipient liver mass increased by 87,101, 119, and 99% at 7, 14, 30, and 60 days after transplantation, respectively. Steatosis did not influence the degree of regeneration or graft function, nor was there a functional difference between grafts of >1% graft to recipient body weight ratio or <1%. CONCLUSIONS: MRI accurately determines right lobe mass. Most liver regeneration occurs in the 1st week after resection or transplantation, and the time course does not differ significantly in donors or recipients. The mass of the graft or remnant segment affects the duration of the regeneration process, with a smaller initial liver mass prolonging the course. Steatosis of <30% had no bearing on liver function or regeneration and, therefore, should not be an absolute criterion for exclusion of donors. A calculated graft to recipient body weight ratio of 0.8% is adequate for right lobe living donor liver transplantation. PMID- 10798758 TI - Sequential histological and immunohistochemical study of the skin of the first human hand allograft. AB - BACKGROUND: On September 1998, the first human hand allograft was successfully performed in Lyon. METHODS: A 48-year-old white man who had suffered accidental amputation of the arm in 1984, received a forearm and hand allograft from a 42 year-old white male cadaveric heart-beating donor. Immunosuppressive therapy included prednisone, mycophenolate mofetil, FK506, and antithymocyte globulins. Sequential skin biopsies were taken from the grafted limb and examined (immuno)histologically to detect a possible graft rejection and to evaluate the structural integrity of the skin of the allograft. RESULTS: The skin showed histologically a normal appearance, except on days 57 and 63, when a mononuclear perivascular cell infiltrate was observed in the dermis; this appeared concomitantly with erythematous lesions of the skin that developed after a slight decrease of the immunosuppressive treatment. These changes were considered as signs of graft rejection, and were reversed by an increase of the immunosuppressive treatment. No skin necrosis was seen at any time. Immunohistochemically, the main cell types of the skin were present throughout the study. From day 77 onward the epidermis of the grafted hand harbored some epidermal Langerhans cells of recipient's origin. CONCLUSION: This study shows that the skin of the hand allograft maintains overall a normal histological structure and contains most essential cell types, including cells of recipient origin, such as Langerhans cells. Furthermore, it shows that in this system of composite tissue transplantation, skin biopsies may reveal a starting graft rejection, before the appearance of clinically obvious lesions. PMID- 10798759 TI - An interleukin-2-IgG-Fas ligand fusion protein suppresses delayed-type hypersensitivity in mice by triggering apoptosis in activated T cells as a novel strategy for immunosuppression. AB - BACKGROUND: Cell-mediated immune responses can be down-regulated by induction of apoptosis of immunoreactive lymphocytes. In the present study, we have tested the feasibility of a strategy for immunosuppression by the selective induction of apoptosis in activated, interleukin (IL)-2 receptor-positive lymphocytes, using a triple IL-2-IgG-FasL fusion protein. The IL-2-IgG-FasL fusion protein combines IL 2 for the selection of activated T cells, with the extracellular domain of the FasL molecule for inducing T-cell apoptosis. These components were separated by the Fc part of IgG1 serving as a spacer as well as for half-life prolongation. METHODS: The gene for the chimeric protein was created by fusing DNA sequences encoding for the three functional components: human IL-2, the Fc part of human IgG1, and the extracellular domain of murine FasL. When the fusion gene was expressed in murine J558L cells, we obtained soluble dimeric immunoglobulin-like proteins in the supernatant. After analyzing the function of the IL-2 and FasL portions individually in vitro, a delayed-type hypersensitivity (DTH) reaction to sheep red blood cells as model for cell-mediated immune responses was investigated to evaluate the IL-2-IgG-FasL-mediated immunosuppression in vivo. RESULTS: In vitro, the IL-2-IgG-FasL fusion protein supported IL-2-dependent proliferation of Fas-resistant CTLL-2 cells, whereas concanavalin A-T blasts were induced to undergo apoptosis by the FasL portion. In vivo, this fusion protein potently inhibited a murine DTH. This was associated with an increased rate of apoptosis in activated lymphocytes in the spleen, even at very low doses of the fusion protein. Furthermore, a second antigen challenge 10 days after IL-2-IgG FasL treatment still failed to elicit a DTH response. CONCLUSION: The abrogation of a standard T cell-dependent immune response in vivo demonstrates that IL-2-IgG FasL can be successfully exploited to trigger the death of deleterious T cells, presenting a potentially useful strategy in the management of autoimmune diseases and allotransplant rejections. PMID- 10798760 TI - Size reduction of small bowels from adult cadaveric donors to alleviate the scarcity of pediatric size-matched organs: an anatomical and feasibility study. AB - BACKGROUND: Small bowel transplantation in children weighing less than 10 kg is hindered by the lack of size-matched donors. The ability to create reduced size small bowel grafts from adult cadaveric donors suitable for use in young children has been studied. METHODS: Volumetric assessment of computed tomography scans were used to evaluate abdominal cavity and small bowel volumes in children. Small bowels were retrieved from adult cadaveric donors and reduced in size. RESULTS: Computed tomography studies of the abdominal cavity showed that the mean volume available for a small bowel graft was 260 ml in children less than 5 kg (n = 5) and 460 ml in children weighing 5-10 kg (n = 5). Fifteen small bowels were successfully reduced to provide an ileal graft of one meter while keeping the whole length of the superior mesenteric artery and vein after their dissection in the proximal part of the mesentery. The mean volume of the grafts created was 270 ml in seven thin patients (body mass index [BMI] <25), 390 ml in five preobese patients (25< BMI<30), and 490 ml in three obese patients (BMI>30). Mesenteric transillumination in thin donors allowed safe dissection and complete hemostasis. No diameter reduction was required. Technical modifications permitted the creation of two grafts, one ileal and the other jejunal from a single donor. Volumetric and surgical data show that implantation of up to two meters of ileum from a thin adult weighing up to 80 kg is feasible in children weighing less than 10 kg. CONCLUSION: Size reduction of adult cadaveric small bowels can provide suitable grafts for children of less than 10 kg and could expand the potential pool of donors for these patients. PMID- 10798761 TI - Prediction of early renal graft function by the measurement of donor urinary glutathione S-transferases. AB - BACKGROUND: We have investigated the possibility of urinary alpha- and pi class glutathione S-transferases (GST-a; GST-pi) serving as a valuable parameter to predict early graft function after transplantation. METHOD: Urinary GST concentrations of 61 donors (DON) and recipients (REC) were analyzed at preoperative, intraoperative, and postoperative periods. We grouped recipients according to the early postoperative graft recovery days. RESULTS: The donor graft function, represented by the donor urinary GST concentration (GST-pi:17,1+/ 12 microg/l mmol creatinine (crea); GST-a:14,3+/-10 microg/mmol crea), sustained a loss in comparison to the healthy controls (GST-a; pi< or =1 microg/mmol crea). According to statistical analysis, the donor GST-pi level showed a strong correlation with graft recovery days-pi (r = 0.84; P<0.001). The early graft function cannot be predicted by means of cold ischemia time (22.8+/-3.4 hr), nor handling time (42.4+/-11.1 min), nor even the intraoperative enzyme concentrations. The GST-pi cut off level (12.55 microg/mmol crea) might predict the possible posttransplant graft dysfunction. The discriminative analysis showed that using only DON GST-pi alone could discriminate well between the groups among all grafts in 68%. CONCLUSION: Prognosis is poorer if the donor GST-pi concentration is above 12.55 microg/mmol crea. On the basis of the determination of GST-pi concentration in the donor urine, we can predict graft viability before the surgical procedure with a reliability of 68%. PMID- 10798762 TI - Lamivudine for hepatitis B in liver transplantation: a single-center experience. AB - BACKGROUND: Liver transplantation for hepatitis B virus (HBV) has been associated with a high rate of reinfection and graft failure. Lamivudine, a potent inhibitor of HBV replication, has been shown to prevent viral recurrence after transplantation. METHODS: The effectiveness of lamivudine monotherapy for the management of HBV recurrence after liver transplantation was assessed. Lamivudine was used in three patient groups: (1) patients started before transplantation and continued after transplantation (n = 13); (2) patients treated after transplantation (n = 15); and (3) patients with de novo hepatitis B after transplantation (n = 4). RESULTS: Median follow-up on lamivudine was 24 months. Active viral replication (HBV-DNA+) was seen in 17 (53%) of 32 at treatment initiation. All lost HBV-DNA at a mean of 2.4+/-1.6 months after lamivudine initiation. Twenty-six (81%) patients remain free of viral recurrence. Six (19%) patients have evidence of breakthrough infection with the YMDD mutant of HBV, two of whom progressed to graft failure. All four patients in group 1 who developed breakthrough had evidence of hepatitis B surface antigen expression in the explanted liver by immunohistochemistry despite being serum HBV-DNA negative before transplantation. No difference was observed among the three groups in DNA clearance or breakthrough rates. CONCLUSIONS: Lamivudine achieves viral DNA clearance in almost all patients. Expression of viral antigens in the liver seems to identify patients at risk of developing HBV-DNA recurrence. Disease-free survival of 81% at 22 months is similar to data with hepatitis B immunoglobulin therapy. Given the safe clinical profile and high efficacy in the prevention of disease recurrence, lamivudine will favorably change the outlook of liver transplantation for HBV. PMID- 10798763 TI - Cyclosporine, FK506, mycophenolate mofetil, and prednisolone differentially modulate cytokine gene expression in human airway-derived epithelial cells. AB - BACKGROUND: The immunosuppressive effects of cyclosporine (CsA), tacrolimus (FK506), mycophenolate mofetil (MMF), and prednisolone in cells from the immunological compartment are well documented. In contrast, limited information is available with respect to the effects of these immunosuppressive drugs on airway-epithelial cells, although these cells may contribute to the development of obliterative bronchiolitis (OB) through the production of interleukin (IL)-6 and IL-8. METHODS: We studied the production of IL-6 and IL-8 proteins by airway derived epithelial cell lines and primary epithelial cell cultures obtained from lung brushings. Transcriptional mechanisms were detected by transient transfections. RESULTS: We demonstrate that CsA dose dependently induces the production of the proinflammatory cytokines IL-6 and IL-8 in both cell lines and primary epithelial cells. FK506 and MMF were also able to upregulate IL-8, although the effect was less dramatic than observed for CsA. Low concentrations of prednisolone (0.01 and 0.001 microg/ml) enhanced IL-6 and IL-8 secretion, whereas concentrations > or =0.01 microg/ml significantly diminished IL-6 secretion. Furthermore, we showed that CsA and prednisolone mediate their effects at the transcriptional level. CONCLUSIONS: The data provide evidence that relevant concentrations of CsA and MMF in vivo may enhance the inflammatory processes in the lower airways of patients after lung transplantation. PMID- 10798764 TI - Pretransplant dialysis status and outcome of renal transplantation in North American children: a NAPRTCS Study. North American Pediatric Renal Transplant Cooperative Study. AB - BACKGROUND: There are no large studies of the effect of pretransplant dialysis status on the outcome of renal transplantation (Tx) in children. This study evaluated the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) registry data for the outcome of Tx in pediatric patients who either (1) received their transplants preemptively or (2) were maintained on dialysis before receiving their transplants. METHODS: We compared graft survival and patient survival rates, incidence of acute tubular necrosis (ATN), acute rejection episodes, and causes of graft failure in peritoneal dialysis (PD) patients with those maintained on hemodialysis (HD) and those undergoing preemptive Tx (PTx). RESULTS: Primary Tx was performed in 2495 children (59% male; 61% Caucasian; 1090 PD, 780 HD, 625 PTx) between 1/1/1992 and 12/31/1996. The overall graft survival rates of the PD and HD groups were similar, but were less than that of the PTx group (3-year: 82% PD and HD, 89% PTx, overall P = 0.0003). Improved graft survival in the PTx group was present only in recipients of grafts from living donors. There was no difference in the overall patient survival rate at 3 years, or in time to first acute-rejection episodes in the three groups. The incidence of ATN in the first 7 days post-Tx was higher in PD and HD patients than in PTx patients (11% PD and 12% HD vs. 2% PTx, P<0.001; HD vs. PD, P = NS). The major single cause of graft failure in each group was: PD, vascular thrombosis (200%); HD, chronic rejection (27%); PTx, acute and chronic rejection (21% each). CONCLUSION: NAPRTCS data show that graft survival is improved in patients receiving PTx, compared with those receiving PD and HD. Graft loss resulting from vascular thrombosis is more common in children who receive PD than in those receiving HD. PMID- 10798765 TI - The experimental (in vitro) and clinical (in vivo) immunosuppressive effects of a rat IgG2b anti-human CD2 mAb, LO-CD2a/BTI-322. AB - BACKGROUND: CD2 is a cell surface glycoprotein expressed on most human T cells and natural killer (NK) cells, working as a cell adhesion and costimulatory molecule. The aim of this paper is to analyze the mechanism of action of a rat IgG2b anti-human CD2 monoclonal antibody (mAb) (LO-CD2a/BTI-322 mAb), which is a potent immunosuppressive agent and inducer of cell death. In vivo, this mAb is able to prevent or treat kidney allograft rejection. METHODS: The mechanisms by which the LO-CD2a/BTI-322 mAb is able to induce inhibition of cell activation and cell death were analyzed by mixed lymphocyte reactions and by flow cytometry. After in vivo treatment, levels of circulating mAb were measured by ELISA as well as anti-rat immunization and cytokine release. RESULTS: We show that the inhibition of cell activation induced by LO-CD2a/BTI-322 mAb after allogeneic or OKT3 stimulation is due to an Fcgamma receptor-dependent CD2 down-modulation and to T-cell depletion through an antibody-dependent cell-mediated cytotoxicity mechanism mediated by NK cells or activated monocytes. Peripheral T- and NK-cell depletion was observed after in vivo treatment with LO-CD2a/BTI322. Cytokine release (TNFalpha) was correlated with some side effects, but only after the first injection, and the effects were never severe or life threatening. CONCLUSION: The correlation between the in vitro and in vivo data suggests that T cell depletion, especially of activated cells, and inhibition of cell activation after CD2 down-modulation are the main mechanisms of action of the LO-CD2a/BTI 322 mAb. PMID- 10798766 TI - Susceptibility of chacma baboons (Papio ursinus orientalis) to infection by hepatitis B virus. AB - BACKGROUND: Because baboons are being considered as a source of xenografts for human liver transplantation in patients with hepatitis B virus- (HBV) induced cirrhosis to forestall infection of the graft by the virus, we undertook a study to ascertain if baboons are resistant to HBV infection. METHODS: Six chacma baboons were inoculated with serum containing HBV and were followed for 52 weeks to detect transmission of infection. RESULTS: Anti-HBc was detected in the serum of four baboons 16 weeks after inoculation. Virions, small spherical particles, and tubular forms were seen at this time in the serum of the one baboon studied by transmission electron microscopy. HBV DNA was detected by polymerase chain reaction in the serum of the same four baboons throughout the period of follow up, as well as in liver tissue obtained after 52 weeks. The specificity of the DNA was confirmed by Southern hybridization. Nucleotide sequences showed complete sequence identity between the HBV DNA in each of the baboon sera and one of the two HBV genotypes inoculated. Serum transaminase levels tested at 4-weekly intervals were always normal and histological examination of liver tissue after 52 weeks showed no evidence of chronic hepatitis. Examination of squash preparations of liver tissue by electron microscopy in one baboon revealed core like particles. CONCLUSIONS: Chacma baboons are susceptible to HBV infection and appear to develop a chronic carrier state. The use of xenografts from baboons should preferably be avoided, but if they are used again for HBV-infected patients it would be prudent to treat the patients as if they had received an organ from a human donor. PMID- 10798767 TI - Decreased donor-specific cytotoxic T cell precursor frequencies one year after clinical lung transplantation do not reflect transplantation tolerance: a comparison of lung transplant recipients with or without bronchiolitis obliterans syndrome. AB - BACKGROUND: Decreased in vitro T cell alloreactivity, demonstrated by decreased frequencies of peripheral blood donor-specific T cell precursors, may reflect a tolerant state after transplantation and lower the risk for development of chronic graft dysfunction. It is unknown whether a decrease in donor-specific T cell frequencies also occurs after clinical lung transplantation and if such a decrease lowers the risk for bronchiolitis obliterans syndrome (BOS), a hallmark of chronic graft dysfunction. Therefore, we compared changes in posttransplant donor-specific cytotoxic T lymphocyte (CTLp) and helper T lymphocyte precursor (HTLp) frequencies in lung allograft recipients with good graft function and in recipients with BOS. METHODS: Donor and third party specific CTLp and HTLp frequencies were determined by limiting dilution assay in pre- and posttransplant (1 year) peripheral blood samples of lung allograft recipients with good graft function (n = 13) and BOS (n = 10). RESULTS: In recipients with good graft function, mean donor-specific CTLp frequencies decreased after transplantation (183 vs. 16 precursors before and after transplantation, respectively). Additionally, HTLp frequencies decreased but this was not specific for donor alloantigens because third party-specific HTLp frequencies decreased also. Surprisingly, recipients with BOS also showed a decrease in mean donor-specific CTLp frequencies after transplantation (332 vs. 49 precursors before and after transplantation, respectively). Again, HTLp frequencies decreased nonspecifically. CONCLUSIONS: We conclude that donor-specific CTLp frequencies decrease after lung transplantation, but that this does not result in transplantation tolerance protecting the lung against the development of chronic graft dysfunction. PMID- 10798768 TI - A dendritic cell line genetically modified to express CTLA4-IG as a means to prolong islet allograft survival. AB - BACKGROUND: Dendritic cells are potent antigen-presenting cells that bind allogeneic T cells. They are thus candidates for targeting immunoregulatory molecules to the alloreactive T cell compartment and suppressing the alloimmune response. METHOD: A dendritic cell line derived from the BALB/c mouse (H2d) was genetically modified to express the immunoregulatory molecule CTLA4-Ig. The ability of these dendritic cell transfectants to downregulate the alloimmune response was tested in an islet transplant model. Allogeneic C57Bl/6 (H2b) mice were rendered diabetic with streptozocin, and they received BALB/c islet (H2d) transplants. Mice were administered 25 million untransfected or CTLA4-Ig transfected D2SC/1 cells i.v. on the day of islet transplantation and 6 days later[fnc]. RESULT: Mice treated with CTLA4-Ig-transfected D2SC/1 cells demonstrated prolonged allograft survival (mean = 20 days, median = 17 days, SD = 9.39) compared with mice treated with untransfected D2SC/1 cells (mean = 12 days, median = 11 days, SD=2.74) or untreated control mice (mean = 11 days, median = 11 days SD = 1.41). Third party allograft survival was not prolonged in mice receiving similar treatment. CONCLUSIONS: These results demonstrate that a genetically modified dendritic cell line can suppress the alloimmune response and prolong islet allograft survival in an allospecific manner. The findings also suggest that genetically modified dendritic cells may be useful in targeting alloreactive T cells and prolonging allograft survival. PMID- 10798769 TI - The induction of specific pig skin graft tolerance by grafting with neonatal pig thymus in thymectomized mice. AB - BACKGROUND: Xenogeneic donor-specific tolerance can be induced by transplanting fetal pig thymus and liver tissue (FP THY/LIV) to thymectomized (ATX), T/NK cell depleted mice. By using neonatal pig tissue, we hoped to overcome two obstacles that arise with the use of fetal pig tissue: (1) the inability to keep fetal pigs alive after harvesting their thymic tissue, resulting in unavailability of their skin or other organs for grafting; and (2) the limited fetal thymic tissue yield, making application to large animals and humans more difficult. METHODS: Neonatal pig thymus tissue (NP THY) was grafted into ATX, T/NK cell-depleted, 3Gy whole body-irradiated, originally immunocompetent B6 mice to evaluate the ability of NP THY to reconstitute mouse CD4+ T cells and to induce xenogeneic tolerance to donor pig skin grafts. RESULTS: Repopulation of mouse CD4+ T cells in the peripheral tissues was observed in T/NK cell-depleted, ATX B6 mice that received NP THY with or without neonatal pig spleen (NP SPL), but not in those receiving NP SPL alone, indicating that pig thymus grafting was necessary and sufficient for mouse T cell recovery. Seven of nine NP THY/SPL-grafted ATX mice and two of six NP THY-grafted ATX mice that reconstituted >5% CD4+ cells in PBL accepted donor pig skin long-term without lymphocyte infiltration, whereas they rejected allogeneic BALB/c skin and third party pig skin grafts as rapidly as euthymic mice. CONCLUSIONS: NP THY can support the development of mouse CD4+ T cells that are functional and specifically tolerant to donor pig antigens in ATX, T/NK cell depleted, 3 Gy whole body-irradiated, originally immunocompetent B6 mice. Additional grafting of NP SPL with NP THY improves the efficiency of tolerance induction in this model. PMID- 10798770 TI - Induction of hyperacute rejection of skin allografts by CD8+ lymphocytes. AB - BACKGROUND: Second-set rejection is generally regarded as a phenomenon mainly mediated by humoral cytotoxic antibodies, although a few discordant data have been presented. In the reported experiments, we have taken advantage of the absence of production of specific cytotoxic alloantibodies contrasting with the normal development of transplantation cellular immunity, in two murine models: chimeric mice and RAG mice. METHODS: Chimeras (BALB/c-->CBA) were obtained by transplantation of 2x10(7) fetal liver cells from BALB/c (H-2d) mice to lethally irradiated CBA (H-2k) mice. After hyperimmunization with third-party C57/ BL6 (B6) (H-2b) skin transplants and with injections of 2x10(7) B6 spleen cells, antibody production, and skin graft survival were analyzed. To identify further the factors or cells responsible for accelerated rejection of B6 skin transplants in hyperimmunized chimeras, transfer experiments were carried out involving the injection of serum, whole spleen cells, spleen T cells, spleen CD8+ T cells or spleen CD4+ T cells from chimeras into BALB/c mice that had received 6 Gy irradiation. The recipient mice were then grafted with B6 skin. Similarly, the immunodeficient RAG mice were used to construct a model of recipient animals with anti-H-2d hyperimmunized B6 T cells in the total absence of antibody. RESULTS: In chimeras, anti-B6 cytotoxic antibodies were not detectable in any of hyperimmunized chimeric mice, yet accelerated rejection of B6 skin transplant occurred: a graft survival of 8.6+/-0.5 days (d), comparable to 8.9+/-0.8 d survival in CBA control mice subjected to the same hyperimmunization procedure, and significantly shorter than that in nonhyperimmunized (BALB/c-->CBA) chimeras (11.6+/-0.5 d) or in non-hyperimmunized CBA control mice (12.1+/-0.6 d). High titers of anti-B6 cytotoxic antibodies were present in the serum of hyperimmunized CBA control mice. In transfer experiments, the graft survival was over 14 d in mice treated with irradiation alone, with irradiation + serum or with irradiation + CD4+ T cells. It was significantly shorter in mice treated with irradiation + whole spleen cells, with irradiation + T cells or with irradiation + CD8+ T cells (8.9+/-0.8 d). Similarly, in immunodeficient RAG mice, reconstitution of the T cell compartment with T cells from hyperimmunized B6 mice led to accelerated rejection of BALB/c skin allografts (11.4+/-1.1 d vs. 18.8+/ 0.8 d when T cells were provided by nonimmunized mice). In a second transfer of cells from these reconstituted RAG mice into naive RAG mice, CD8+ T cells were shown to induce accelerated rejection of skin allografts (12.0+/-0.6 d) whereas CD4+ T cells were much less efficient (16.5+/-0.1 d). CONCLUSION: These data indicate that T cells, and especially the CD8+ subset, can be responsible for second-set rejection in the absence of anti-donor antibodies in chimeric and RAG mouse models. These sensitized CD8+ T cells are also likely to play an important role in normal mice, in addition to that of cytotoxic antibodies. PMID- 10798771 TI - Transhepatic neutrophil and monocyte activation during clinical liver transplantation. AB - BACKGROUND: During experimental liver transplantation, neutrophil sequestration results in increased oxygen free radical production and correlates inversely with graft viability. Neutrophil activation in clinical liver transplantation is poorly understood. METHODS: We assessed leukocyte sequestration and transhepatic differences of neutrophil and monocyte CD11b expression, neutrophil free radical production, and plasma concentrations of interleukin 6 and interleukin 8 in nine patients during liver transplantation. RESULTS: Significant hepatic neutrophil sequestration occurred during initial graft rewarming with portal blood, after inferior vena cava declamping, and after hepatic artery declamping (all P<0.05). A positive transhepatic difference (i.e., outcoming - ingoing) in CD11b expression of neutrophils was observed after portal vein declamping (51+/-32 relative fluorescence unit [RFU]) and in CD11b expression of monocytes during initial graft rewarming (67+/-86 RFU, both P<0.05). A transcoronary increase in both unstimulated (74+/-80 RFU) and N-formyl-methionyl-leucylphenylalanine stimulated (112+/-168 RFU) neutrophil free radical production took place after hepatic artery declamping (both P<0.05). A negative transcoronary difference of interleukin 6 occurred during initial graft rewarming (-192+/-176 pg/ml) and a positive difference of interleukin 8 occurred after hepatic artery declamping (17+/-23 pg/ml, both P<0.05). CONCLUSIONS: Hepatic sequestration and transhepatic activation of neutrophils, and hepatic production of interleukin 8 occur during clinical liver transplantation. A splanchnic influx of interleukin 6 occurs to the graft, possibly modulating neutrophil-mediated graft reperfusion injury. PMID- 10798772 TI - Implantation of rat insulinoma cell line into cyclosporine treated rats. Effect of the in vivo environment on beta-cell specific gene expression. AB - BACKGROUND: Transplantation of engineered beta cell-derived lines is a promising modality for cell-based therapy of diabetes mellitus. The in vivo environment and antirejection and other medications may have significant effects on the differentiation and proliferation of the transplanted beta cells, thus affecting their function. The effect of the in vivo environment on expression of genes encoding proteins involved in insulin production, secretion, and glucose sensing were analyzed in the RIN 104638 cell line. METHODS: RIN 104638 cells, were used for s.c. implantation in cyclosporine treated rats and for parallel in vitro culture. The differential expression of the insulin, PDX-1, GLUT-2, and glucokinase genes were assessed by quantitative reverse transcription polymerase chain reaction. RESULTS: The in vivo environment of cyclosporine-treated rats, preserved most of the differentiated characteristics of the implanted cells. Insulin and glucokinase gene expression were maintained at high levels, although GLUT-2 expression decreased. This was in contrast to the substantial decrease of all the three genes expression when cultured in vitro. Cyclosporine treatment reduced insulin and GLUT-2 gene expression in in vitro culture. CONCLUSIONS: Beta cell implantation in cyclosporine-treated rats induces alteration in expression of genes pivotal to insulin production and secretion and the glucose sensing abilities. The normal in vivo environment improves the implanted b cell function by increasing the insulin gene expression and content. Furthermore, it reverses some of the dedifferentiating changes caused by the in vitro culture. This may have a positive effect on the therapeutic efficiency of this cell line. PMID- 10798773 TI - In vivo modulation of the allogeneic immune response by human fetal kidneys: the role of cytokines, chemokines, and cytolytic effector molecules. AB - BACKGROUND: We have recently demonstrated that human fetal renal tissue, implanted under the kidney capsule of severe immunodeficient rats, escapes early destruction by intraperitoneal infusion of allogeneic human peripheral blood mononuclear cells, compared with the rapid rejection of implants of human adult kidney tissue. Variable amounts of human mononuclear infiltrates were seen in the transplanted fetal kidney, however, prolonged survival of the fetal tissue (maintenance of graft architecture and significant growth) was independent of the cellular infiltrate. METHODS: We have used this experimental model to sequentially analyze transcript levels of interferon-gamma and interleukin (IL)-2 (T helper 1 cytokines), IL-4 and IL-10 (T helper 2 cytokines), RANTES, MIP1beta (beta chemokines) and their receptor CCR5, and Fas ligand (cytolytic effector molecule). Analysis was performed by reverse transcriptase-polymerase chain reaction, in both fetal and adult kidney grafts, after infusion of allogeneic human peripheral blood mononuclear cells. RESULTS: Transcript levels of interferon-gamma and IL-2 in the fetal grafts were markedly reduced throughout follow-up, compared with those observed in the adult implants. Peak levels of these cytokines appeared late in the rejection process. Concomitant with these findings, IL-4 mRNA was up-regulated during the early phase, whereas IL-10 mRNA persisted throughout the rejection process, indicating that a T helper 2 bias occurred in the fetal grafts. In addition, RANTES (after an early peak), MIP1beta, CCR5, and Fas ligand mRNA levels were suppressed in the fetal grafts compared with those in the adult grafts. CONCLUSIONS: These findings indicate that the immune response of kidney rejection is dependent on whether the target organ is of fetal or adult origin, and suggest that an allogeneic immune system mounts a T helper 2-biased response when the target organ is of fetal origin. PMID- 10798774 TI - Histopathological features of hepatitis C in renal transplant candidates [see comment]. AB - BACKGROUND: Although hepatitis C virus (HCV) infection is common in renal transplant candidates, its clinical significance remains unclear in this population. Little detailed information is available about the histological severity of HCV infection in these patients. We evaluated the liver biopsy features of chronic HCV in a large population of renal transplant candidates and investigated associations between histopathological changes and host- and virus related factors. METHODS: Thirty-seven patients seropositive for anti-HCV with chronic renal failure (CRF) referred to UCLA Medical Center for kidney or kidney/liver transplantation during the period 1992-1997 were included. HCV genotype and viral load were measured. A multivariate analysis by logistic regression model was performed: age, gender, race, HCV load and genotype, CRF level, aspartate and alanine aminotransferase activity, duration of HCV infection, underlying nephropathy, and alcohol abuse were independent variables; liver histology score was assumed a dependent variable. RESULTS: Liver disease was present in all HCV-infected patients. Logistic regression analysis revealed that histological damage was (P = 0.0017) independently associated with the CRF level; the severity of liver disease, as shown by univariate analysis, being significantly higher in CRF patients not requiring dialysis than among dialysis population. All patients on dialysis showed mild or moderate necroinflammatory activity; the majority (22/28 = 79%) of these individuals had fibrosis, three (3/28 = 11%) dialysis patients had established cirrhosis. Thirty-one (84%) of 37 patients were tested by polymerase chain reaction, 25 (81%) patients had detectable HCV RNA in serum, the mean HCV load among viremic patients was 10.9x10(5) copies/ ml. The most frequent HCV genotypes were la (8/24 = 33%) and 1b (7/24 = 29%), followed by genotype 2b (3/24 = 12%). CONCLUSIONS: Pathological changes on liver biopsy were observed in all HCV-infected patients awaiting renal transplantation. The severity of histologic damage observed on liver biopsy was less in dialysis than predialysis CRF patients. All dialysis patients had mild or moderate necroinflammatory activity; fibrosis was frequent with 11% of them having cirrhosis. The HCV viral load was rather low; no relationship between liver histology changes and virological features of HCV or aminotransferase activity was apparent. Further studies with repeat liver biopsies after kidney transplantation to observe the evolution of HCV-related liver disease after immunosuppressive therapy are indicated. We suggest including liver biopsy in the evaluation of the HCV-infected renal transplant candidate. PMID- 10798775 TI - Renin-angiotensin system gene expression in post-transplant hypertension predicts allograft function. AB - BACKGROUND: Registry analyses and single-center studies have demonstrated that hypertension significantly increases the risk for chronic graft loss. The graft itself may contribute to posttransplant hypertension, and intragraft vasoactive hormones therefore, may be dysregulated in posttransplant hypertension. METHODS: We used the reverse-transcription polymerase chain reaction to assess the intragraft regulation of renin-angiotensin system transcripts in biopsy samples from 42 stable renal transplant patients with posttransplant hypertension. We also examined mRNA expression of inducible nitric oxide synthase, transforming growth factor-beta (TGF-beta), select cytokines, and metalloproteinase transcripts in biopsy tissue. Polymerase chain reaction products were quantitated using high performance liquid chromatography and normalized to beta-actin mRNA expression. Serum creatinine, glomerular filtration rate or creatinine clearance and tubular atrophy on biopsy were concurrently assessed. RESULTS: Renin and select Thl cytokine mRNA expression correlated with blood pressure. Type 1 angiotensin II receptor mRNA expression significantly correlated with glomerular filtration rate or creatinine clearance (P = 0.034) and inversely correlated with Th1 cytokines, inducible nitric oxide synthase, and cyclooxygenase-1 mRNA expression (P< or =0.013 for each). Type 1 angiotensin II receptor mRNA also approached a significant inverse correlation with TGF-beta mRNA expression (P = 0.09). Conversely, angiotensin-converting enzyme mRNA expression directly correlated with Thl cytokine (P< or =0.008 for each) and TGF-beta mRNA expression (P = 0.006). Type 1 angiotensin II receptor mRNA expression also correlated with matrix metalloproteinase-1 promoter region, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and tissue inhibitor of matrix metalloproteinase-3 mRNA expression. Notably, matrix metalloproteinase-1 promoter region, tissue inhibitor of matrix metalloproteinase-2, and tissue inhibitor of matrix metalloproteinase-3 inversely correlated with TGF-beta mRNA expression (P< or =0.0027 for each). Type 1 angiotensin II receptor mRNA expression at biopsy directly correlated with glomerular filtration rate at 2 year's follow-up. However, angiotensin-converting enzyme mRNA expression at biopsy inversely correlated with glomerular filtration rate at 2 year's follow-up. CONCLUSIONS: These data suggest that allograft-level RAS gene expression may be predictive of future graft function in the setting of diastolic hypertension. PMID- 10798776 TI - A large, single center investigation of the immunogenetic factors affecting liver transplantation. AB - BACKGROUND: Reports on the relevance of immunogenetic factors in liver transplantation are often conflicting or inconclusive. We have, therefore, investigated a range of factors that may underlie liver graft survival. METHODS: The influences of HLA, flow cytometric, and enhanced cytotoxic crossmatching and immunoglobulin (Ig)A levels on graft survival, and acute and chronic rejection were investigated for a single center involving 446 patients over 13 years. RESULTS: The effect of HLA mismatching on graft survival was significant (P<10( 2)) and was reversed in recipients with autoimmune diseases (P<0.5x10(-2)), whereas the effect of HLA mismatches on the level of acute rejection was detrimental in all recipients. There was a significant effect of a positive cytotoxic crossmatch on 3-month (P<10(-5)) and 1-year (P<10(-4)) graft survival, and an additional effect of the flow cytometric crossmatch was seen for chronic rejection (P<10(-2)) and acute rejection (P<10(-2)). Recipients with HLA A1,B8,DRB1*0301 had higher levels of acute rejection (P<0.5x10(-2)), and recipients who received an ABO compatible-nonidentical transplant have a significantly higher risk (P<10(-2)) of developing chronic rejection. Finally, the beneficial effect of high serum IgA and, specifically, IgA anti Fab, seen in renal transplants was not evident in liver transplants, and in fact the opposite may be true, at least for acute rejection (P<0.5x10(-2)). CONCLUSIONS: By separating the recipients with autoimmune disease from other patients and by including acute and chronic rejection as outcome parameters, we have used the power of a large single-centre study to delineate the significance of some of the important immunogenetic factors involved in liver transplantation. PMID- 10798777 TI - Left hemihepatectomy in living donors with a thick middle hepatic vein draining the caudal half of the right liver. AB - Donor safety is the first consideration in living related liver transplantation. Left hemihepatectomy including the middle hepatic vein is a reasonable donor procedure for obtaining a large graft for living related liver transplantation. This procedure, however, needs to be modified in donors with hepatic venous variation. While carrying out donor hepatectomy, we encountered two cases showing a variant form of hepatic venous drainage comprising a thick middle hepatic vein draining segment 6 of the liver. This variation made it necessary to preserve the middle hepatic vein in the donor liver remnant. Failure to recognize such a variant would result in congestion in the remaining right liver of the donor. To guarantee donor safety, evaluation of the drainage area of the corresponding hepatic vein is a matter of great importance in donor hepatectomy. PMID- 10798778 TI - Nasal natural killer cell lymphoma in a post-renal transplant patient. AB - Posttransplant lymphoproliferative disorders in organ allograft recipients are most commonly of B-cell origin and only occasionally of T-cell origin. We present here a case of nasal natural killer cell lymphoma associated with Epstein-Barr virus that occurred in a recipient of a renal transplant 4 years posttransplantation. Immunohistochemically, the lymphoma cells showed CD2-, surface CD3-, cytoplasmic CD3E+, CD56+, CD57-, CD16-, and CD43+ phenotype. Analyses of T-cell receptor beta and gamma genes showed germ line configurations. EBER-1 was detectable in the lymphoma cells. The patient was diagnosed as having natural killer cell lymphoma and was treated with six courses of combination chemotherapy for non-Hodgkin's lymphoma He has been in remission for more than 3 years thereafter. To the best of our knowledge, this is the first report of a posttransplant NK cell lymphoma associated with Epstein-Barr virus. PMID- 10798779 TI - Unmasking of primary hyperaldosteronism by renal transplantation. AB - BACKGROUND: Primary hyperaldosteronism is an uncommon cause of hypertension in the general population. Given the mechanism of action of aldosterone clinical manifestations may not occur in the setting of end stage renal disease. However, if a successful renal transplant is performed clinical manifestations may occur. METHODS: We present a case of a patient with a preexisting adrenal adenoma who only presented with clinical signs of hyperaldosteronism after renal transplantation. Patients' work-up included plasma aldosterone, plasma renin activity, serum cortisol, and estimation of trans tubular potassium gradient. RESULTS: The patient's serum aldosterone was markedly elevated with a relatively suppressed plasma renin activity. Trans tubular potassium gradient was high in the presence of hypokalemia. CONCLUSION: Previously silent hyperaldosteronism may be unmasked by a successful renal transplant. PMID- 10798781 TI - Growth of intramedullary lipoma in a renal transplant recipient. AB - BACKGROUND: Administration of glucocorticoids can lead to a variety of complications in addition to deposition of fat leading to cushingoid features. Corticosteroids, either endogenously produced or exogenously administered, are implicated in the growth of lipomas in different anatomic locations including the epidural space in the spinal cord causing cord compression. METHOD: We report a growth of lipoma in an unusual site in a 28-year-old female renal transplant recipient within 6 weeks of renal transplant surgery. RESULT: Our patient had an intradural lipoma that had merged with the medulla of the spinal cord making its total excision unfeasible without damaging the spinal cord. CONCLUSION: Epidural lipomas causing cord compression is documented in patients receiving long-term corticosteroid therapy. This is the first case of intramedullary lipoma of the spinal cord that may be related to steroid use. PMID- 10798780 TI - Low incidence of kidney rejection after simultaneous kidney-pancreas transplantation after antithymocyte globulin induction and in the absence of corticosteroids: results of a prospective pilot study in 28 consecutive cases. AB - BACKGROUND: Recipients of simultaneous kidney-pancreas transplantation receive a combination of polyclonal antithymocyte globulin (ATG), cyclosporin or tacrolimus, mycophenolate mofetil (MMF) and corticosteroids (Cs). To avoid the side effects and adverse events associated with Cs, we investigated a new immunosuppressive regimen without Cs after simultaneous kidney-pancreas transplantation. METHODS: A total of 28 consecutive patients who underwent simultaneous kidney-pancreas transplantation were included in this study. All patients received ATG, cyclosporin, and MMF. RESULTS: All patients but one tolerated the ATG course well. MMF was definitively discontinued in three patients because of leukopenia. Cytomegalovirus infection was diagnosed in eight patients (28.5%). Only two patients (7%) required an antirejection treatment. Patient, kidney, and pancreas survival is currently 96.4, 96.4, and 75%, respectively. CONCLUSIONS: The combination of ATG, cyclosporin, and MMF, without Cs, was well tolerated. The unexpectedly low (7%) incidence of acute kidney rejection observed suggests that Cs may partially interfere with the immunosuppressive effect of ATG. PMID- 10798782 TI - The clinical impact of early gram-positive bacteremia and the use of vancomycin after allogeneic bone marrow transplantation. AB - BACKGROUND: Gram-positive bacteremia (GPB) is an increasing infection after allogeneic bone marrow transplantation (BMT). Our purpose was to identify risk factors for GPB, to evaluate its impact on early mortality and morbidity, and to compare prophylactic with empirical intravenous vancomycin. METHODS AND RESULTS: We studied 89 consecutive BMTs in adult patients. Early GPB occurred in 29% of posttransplantation episodes. T-cell depletion (odds ratio [OR]: 0.18) and vancomycin-prophylaxis (OR: 0.28) reduced the risk of GPB. Mortality at 6 weeks was not significantly different in patients with GPB (15% vs. 9.5%, P = 0.669). GPB was associated with the development of major complications, the use of amphotericin B, and prolonged neutropenia. Vancomycin prophylaxis led to an increased risk of early renal dysfunction (OR: 18.7). CONCLUSION: GPB contributes to overall morbidity during the early post-BMT episode but has no impact on mortality. Vancomycin prophylaxis is effective to reduce GPB but has a negative effect on renal function. PMID- 10798784 TI - Transforming growth factor-beta and interleukin-10 subvert alloreactive delayed type hypersensitivity in cardiac allograft acceptor mice. AB - We have previously reported that temporary treatment of cardiac allograft recipients with gallium nitrate (GN) results in indefinite graft survival, and the inability to mount donor-reactive delayed type hypersensitivity (DTH) responses. We report that antibodies to either transforming growth factor-beta (TGFbeta) or interleukin-10 (IL10) can uncover DTH responses to donor alloantigens in cardiac allograft acceptor mice. The DTH responses uncovered with TGFbeta-reactive antibodies can be blocked by exogenous IL10, and those uncovered with IL10-reactive antibodies can be blocked by exogenous TGFbeta. These data demonstrate that allograft acceptor mice are fully allosensitized, and poised to make donor-reactive cell-mediated immune responses. However, such responses are subverted by a donor alloantigen-dependent mechanism that involves TGFbeta and IL10, which in turn interfere with local cell-mediated immune responses. PMID- 10798783 TI - The effect of polymorphisms in tumor necrosis factor-alpha, interleukin-10, and transforming growth factor-beta1 genes in acute hepatic allograft rejection. AB - BACKGROUND: The occurrence of acute rejection in orthotopic liver transplantation is unpredictable. The role of cytokines in the process of rejection is not entirely clear. We investigated polymorphisms in the genes encoding tumor necrosis factor (TNF)-alpha, interleukin (IL)-10, and transforming growth factor (TGF)-beta1, which affect the amount of cytokine produced in vitro, in a liver transplant population to determine any association with acute rejection. METHOD: DNA was extracted from whole blood of liver transplant patients. After amplification with polymerase chain reactions, the polymorphisms at TNF-alpha 308, IL-10 -1082, and TGF-beta1 +869 and +915 were determined using sequence specific oligonucleotide probes. Acute cellular rejection was a clinical and histological diagnosis. RESULTS: Acute cellular rejection requiring treatment occurred in 68 (48%) of 144 patients. Acute cellular rejection was significantly associated with the TNF-alpha -308 A/A genotype (P<0.02). There was no significant association with either IL-10 or TGF-beta1 polymorphisms in acute rejection. CONCLUSION: Patients with a homozygous TNF-alpha -308 genotype A/A are more likely to suffer from acute cellular rejection after liver transplantation. PMID- 10798785 TI - STAT 6 up-regulation by FK506 in the presence of interleukin-4. AB - BACKGROUND: FK506 perturbs normal phosphorylation by inhibition of the PP2B protein phosphatase, calcineurin. Calcineurin activity is required for intracellular signal transduction via the T cell receptor that in turn leads to either TH1, or TH2, -type responses to antigen. This choice of response involves differential phosphorylation of STATS (Signal Transducers and Activators of Transcription) for induction of STAT activity. Interferon-gamma activates STAT1, a TH1-type mediator, and interleukin-4 activates STAT6, a TH2-type mediator. We ask if FK506 biases STAT activation toward a TH2-type response. METHODS: Cells of the RAW 264.7 mouse macrophage line were treated with interleukin-4, or interferon-gamma, plus or minus FK506, and any effect on STAT6 and STAT1 was compared. RESULTS: Interleukin-4 specifically induced activation of STAT6, and pretreatment with FK506 enhanced this activity. Interferon-y induced STAT1 activity but this was not influenced by FK506 pretreatment. CONCLUSION: FK506 protects against allograft rejection by inhibiting interleukin-2 production. Such protection may be enhanced by FK506-mediated up-regulation of STAT6 activity. PMID- 10798786 TI - Thiopurine S-methyltransferase gene polymorphism is predictive of azathioprine induced myelosuppression in heart transplant recipients. AB - Azathioprine (AZA) is metabolized via the cytosolic enzyme thiopurine S methyltransferase (TPMT). TPMT activity exhibits genetic polymorphism with four prevalent (75%) mutant alleles TPMT*2 (G238C) and TPMT*3 (A719G and/or G460A) and a wild-type allele TPMT*1. To test the hypothesis that presence of these mutations is associated with greater toxicity of AZA in heart transplant recipients, 30 consecutive patients treated with AZA were followed up for the first month after heart transplant. Mutation of TPMT gene (mutation-specific polymerase chain reaction-based methods) was observed in four patients (A719G: n = 2; A719G plus G460: n = 2). Agranulocytosis did not occur in patients with the wild genotype. It occurred in the two patients with mutation A719G and there was a 40% drop in neutrophils in the two other patients. Discontinuation of AZA in the four mutant patients corrected for the drop. Presence of TPMT mutations is associated with a greater likelihood of agranulocytosis. Determination of these mutations could reduce the risk for hematological side-effects. PMID- 10798787 TI - Tumor necrosis factor-alpha and tumor growth factor-beta1 genotype: partial association with intragraft gene expression in two cases of long-term peripheral tolerance to a kidney transplant. AB - Genomic DNA was obtained from peripheral blood samples of patients JB and DS each of whom received a kidney transplant at 16 years of age from a serologically HLA DR matched and HLA-class I -mismatched donor. Both patients discontinued immunosuppression after 1-2 years and retained good renal function for an additional 5 years or more. DNA was analyzed for genetic polymorphisms in the tumor necrosis factor-alpha (TNFalpha) and tumor growth factor-beta1 (TGFbeta1) loci. Biopsy samples obtained during stable function (DS, JB) and during rejection (JB) were analyzed by RT/PCR for cytokine gene expression. Both patients had a high responder genotype for TGFbeta1. DS had a low responder TNFalpha genotype, while JB and his donor were both genotypically TNFalpha intermediate responders. DS had a high TGFbeta1: TNFalpha mRNA ratio in two biopsies obtained during tolerance, while JB, who eventually lost his graft, had more TNFalpha than TGFbeta1 mRNA. The results suggest a possible role for cytokine immunogenetics in the stability of peripheral tolerance. PMID- 10798788 TI - Management of injuries to the anterior cruciate ligament: results of a survey of orthopaedic surgeons in Canada. AB - OBJECTIVE: To identify the approaches to management of anterior cruciate ligament (ACL) injury by Canadian orthopedic surgeons. METHODS: A questionnaire was mailed to 234 physicians randomly chosen from the Canadian Orthopaedic Association directory to obtain the following information: 1) how orthopaedic surgeons diagnose acute hemarthroses; 2) how patients in any of three common ACL injury scenarios would be managed; 3) what variations exist in surgical technique; and 4) how patient variables such as age, gender, and alignment influence the decision-making process. RESULTS: The return rate was 72%, and 56% of respondents were from academic centers. Patients such as those described in the protocol are routinely managed by 80% of the respondents. The diagnosis of acute hemarthrosis is predominantly made by means of clinical examination and radiographs. Magnetic resonance imaging (MRI) is used occasionally by 43% and routinely by 6% of those who responded; arthroscopy is used routinely by 24%. For the competitive athlete with a complete ACL tear, 64% would recommend reconstruction and 33% would recommend bracing and rehabilitation. For reconstruction, 59% would use bone patellar tendon-bone (B-PT-B) autograft and 32% would use hamstring tendon autograft; 40% would incorporate the ACL stump during reconstruction. Of the respondents, 77% would advocate ACL reconstruction for competitive athletes with chronic ACL injury. Of these, 63% would use B-PT-B autograft and 27% would use hamstring tendons. If bracing and rehabilitation failed, 98% would recommend ACL reconstruction. In ACL reconstruction, synthetic augmentation would be used by 12% in chronic cases and by 16% in acute cases. In making the decision to perform ACL reconstruction, 53% consider limb alignment to be important and 67% consider moderate patellofemoral pain to be important. Seventy-one percent are influenced by patellofemoral pain when choosing a surgical technique, with a trend toward semitendinosis autograft rather than B-PT-B autograft reconstruction. For the 8 year-old child with an acute ACL injury, 63% of the respondents would recommend rehabilitation and bracing. For the 14-year-old, 45% would recommend rehabilitation and bracing and 37% would recommend ACL reconstruction after physeal closure. CONCLUSION: The results of the survey indicate that, with respect to some of the issues, there is a wide variation in management of acute and chronic ACL injuries among Canadian orthopedic surgeons. Future research and randomized, controlled clinical trials should be directed toward these areas. PMID- 10798789 TI - The relationship between lower extremity injury, low back pain, and hip muscle strength in male and female collegiate athletes. AB - OBJECTIVE: To determine the relationship of previous lower extremity (LE) injury and/or low back pain (LBP) on hip abduction and extension strength. DESIGN: Cohort study of college athletes at time of preparticipation screening physical. SETTING: An NCAA Division I college. PARTICIPANTS: Two hundred ten college athletes (140 males and 70 females) from an NCAA Division I school. MAIN OUTCOME MEASURES: Mean and maximal hip abduction and extension strengths were recorded using a specially designed dynamometer anchoring station. Previous injury to the LE or LBP in the past year was recorded via personal interview at the time of screening and verified by review of previous injury records. RESULTS: A significant difference in side-to-side symmetry of maximum hip extension strength was observed in female subjects who reported LE injury or LBP as compared to those who did not. Side-to-side difference in hip strength, however, did not differ between male athletes, regardless of reported LE injury or LBP status. CONCLUSION: Female athletes appear to have a differing response of the proximal hip musculature to LE injury or LBP, as compared with their male counterparts. Research is under way to further validate these findings. CLINICAL RELEVANCE: This study provides some reasoning to support the screening of hip strength during the preparticipation physical, as it may be important in the prevention of LE injury and LBP in collegiate athletes. PMID- 10798790 TI - Can 16-18-year-old elite ballet dancers improve their hip and ankle range of motion over a 12-month period? AB - OBJECTIVE: To determine the effect of a 12-month intensive ballet training regimen on hip and ankle range of motion in male and female, first- and second year professional dancers. DESIGN: 12-month longitudinal follow-up. SETTING: National classical ballet school in Australia. PARTICIPANTS: 28 female and 20 male full-time ballet students with a mean +/- 1 SD, ages 16.8 +/- 0.8 and 17.7 +/- 1.2 years, respectively. MAIN OUTCOME MEASURES: Degrees of range of motion of left and right sides for the following movements: standing plie in parallel passive ankle dorsiflexion (DF); standing turnout in the balletic first position- lower leg external rotation (LLER); supine hip external rotation (ER); supine hip internal rotation (IR). An additional range of motion was calculated: external rotation below the hip joint (BHER) derived by subtracting hip ER from LLER. MAIN RESULTS: In all subjects combined, hip and ankle ranges increased statistically on the right. However, the amount was generally minimal and most at the borderline of the amount of error associated with the measurement tool. While there was no change in LLER, there was a decrease in BHER. There were no overall gender differences, and year differences existed only for left hip ER and total hip ER with first-year dancers showing significant improvements in these ranges. For DF and sum of hip IR, first-year males and second-year females had increases in range. There was a negative relationship between baseline range and the amount of change over the 12 months. CONCLUSIONS: Dancers ages 16-18 years who enter full-time ballet training did not augment their ankle dorsiflexion to any appreciable degree. Some, but certainly not all, increased their hip active external rotation over 12 months without increasing their total lower limb turnout. Hip ER was more likely to improve in the first-year rather than second year student in this elite full-time training school. PMID- 10798791 TI - The acute neuropsychological effects of heading in soccer: a pilot study. AB - OBJECTIVE: The main objective of this study was to prospectively examine the acute effects of heading in soccer on cognitive function. DESIGN: This was a prospective cross-over study using a brief neuropsychological battery to assess cognitive function. The tests were performed before and after two separate practice sessions, with athletes serving as their own controls. SETTING: Male and female Division I college athletes. PARTICIPANTS: Members of the men's and women's varsity collegiate Penn State University soccer teams. Forty-four males and 56 females entered and finished the study. All athletes had a normal physical examination. INTERVENTIONS: Before and after both practice sessions, all athletes had a brief battery of neuropsychological tests and a symptom checklist. MAIN OUTCOME MEASURES: Neuropsychological tests symptom checklist compared at baseline with those after the practice sessions. RESULTS: There were no significant differences in pretest scores between groups and no difference on posttest scores between heading and nonheading groups. A significant difference was detected using MANOVA (p = < 0.001) between pre- and posttest scores for measures of attention and concentration, indicating a practice effect. A gender-specific effect in one test measuring attention and concentration was found. There was no difference in symptoms before and after heading as compared with exertional controls. CONCLUSIONS: In this study, soccer players heading the ball does not appear to lead to acute changes in cognitive function as assessed by a brief neuropsychological battery. There are practice effects that occur with repetitive neuropsychological testing and gender differences with certain tests. PMID- 10798792 TI - High school cross country running injuries: a longitudinal study. AB - OBJECTIVE: To determine the incidence rate of injury among high school cross country runners over a 15-year period. DESIGN: Prospective-longitudinal. SETTING: Twenty-three high schools in western Washington State under the surveillance of the University of Washington Athletic Health Care System between 1979-1994. PARTICIPANTS: One hundred and ninety-nine cross country teams. MAIN OUTCOME MEASURE: Injuries resulting from running in a cross country practice or meet. RESULTS: There were 1,622 injuries for an overall injury rate of 13.1/1,000 athletic exposures (AEs), i.e., participation of a runner in a practice or meet. Girls had a significantly higher overall injury rate (16.7/1,000 AEs) than boys (10.9/1,000 AEs) (p < 0.0001). Girls also had significantly higher injury rates than boys for both initial (p < 0.0001) and subsequent injuries (p < 0.0001), especially those at the same body location (p = 0.0001). This difference in risk estimates was consistent over a 15-year period. Nearly three-fourths of the injuries resulted in < or =4 days of disability. Overall, higher rates of initial injuries were reported during practices (9.2/1,000 AEs) than in meets (7.8/1,000 AEs) (p = 0.04). Shin injuries had the highest overall rates of new injury (1.9/1,000 AEs) and reinjury at the same body location (53.9/1,000 AEs). Girls had significantly higher initial injury rates than boys for shin (p < 0.0001), hip. and foot injuries (p < 0.01), and higher reinjury rates for knee. calf, and foot injuries, respectively (p < 0.05). CONCLUSION: The results of this study suggest that girl cross country runners are at higher risk of injury and reinjury than boy cross country runners. PMID- 10798793 TI - Safety recommendations in Shotokan karate. AB - OBJECTIVE: To study risk factors for injury in karate and to establish safety recommendations. DESIGN: Cross-sectional survey of karate injuries. SETTING: Shotokan karate clubs in Boston, Massachusetts, Dallas, Texas, and Winnipeg, Manitoba, Canada. PARTICIPANTS: All athletes training at each club received surveys. A total of 114 surveys were analyzed (74% response rate). MAIN OUTCOME MEASURES: Presence of injuries (requiring any time off from practice), major injuries (requiring at least 7 days off), and multiple injuries (3 or more injuries). RESULTS: No statistically significant differences were found with respect to sex. For all outcomes, karateka younger than 18 years of age had fewer injuries. The number of karateka with injuries and with multiple injuries increased with belt rank until brown belt, then reached a plateau. Brown and black belts had a greater frequency of major injuries than the lower ranks. Training more than 3 hours per week correlated with an increase in injuries, major injuries, and multiple injuries. CONCLUSION: Shotokan karate appears to be a safe sport, especially for those younger than 18 years of age. Risk of injury increases significantly when younger karateka of any rank or older karateka of lower ranks train more than 3 hours per week; therefore, to reduce the risk of injury to less than 50%, weekly training should be limited to a maximum of 3 hours in these groups. PMID- 10798794 TI - Comparative effects of zopiclone and loprazolam on psychomotor and physical performance in active individuals. AB - OBJECTIVE: Sedative hypnotics are used by athletes to alleviate precompetition anxiety and insomnia. The effects of these agents on exercise tolerance have not been extensively researched. DESIGN: To determine the effects of sedative hypnotics on psychomotor and physical performance, a double-blind, placebo- (P) controlled, cross-over designed trial investigated the effects of zopiclone (Z) and loprazolam (L) on performance in 12 athletes. INTERVENTION: Subjects ingested either P, Z (7.5 mg), or L (2 mg) on three different occasions separated by a 1 week washout period. Eye-hand coordination tests, a 30-m sprint test, an agility test, and a graded treadmill run to exhaustion for determination of VO2max were performed 10 hours after drug administration. RESULTS: Subjects reported a significantly greater hangover effect following ingestion of L (8/11 subjects) compared with ingestion of Z (3/11 subjects; p < 0.01). A greater number of subjects felt alert after ingestion of P (9/11 subjects) and Z (9/11 subjects) compared with L (4/11 subjects; p < 0.01). The results of the eye-hand coordination tests, the 30-m sprint, the T-test, the VO2max, and the time to exhaustion during the treadmill run were not significantly altered following the ingestion of P, Z, and L. There was a significant difference between the delta values for Z and L for the number of missed responses in the eye-hand coordination tests (p < 0.02). Therefore, following the ingestion of L, subjects experienced a significant hangover effect and altered reaction time, whereas the ingestion of Z did not significantly impair either psychomotor or physical performance in the administered tests. CLINICAL RELEVANCE: Investigate the extent of the effects of sedative hypnotics on exercise performances, enabling team physicians to prescribe such drugs to the athlete more effectively. PMID- 10798795 TI - Changes in muscle power and neuromuscular efficiency after a 40-minute downhill run in veteran long distance runners. AB - OBJECTIVE: To establish whether there was a relationship between the total accumulated distances of racing in veteran runners and the neuromuscular efficiency of the quadriceps muscles before and after a downhill run. SETTING: University of Cape Town, Sports Science Institute of South Africa. PARTICIPANTS: Twenty male veteran long distance runners (45-50 years of age) with a range of training (1,300 km to 111,280 km) and racing (0 km to 9,737 km) experience. INTERVENTION: A 40-minute downhill run (-10% decline) on a treadmill, at a speed corresponding to 70% of the subject's peak treadmill running speed. MAIN OUTCOME MEASURES: The difference between integrated electromyography (IEMG)/mean force over a 5 s maximal voluntary isometric contraction before and after the downhill run was calculated as the delta (delta) neuromuscular efficiency. This was related to the total kilometers trained, current training distance, total kilometers raced, and number of races > 56 km. The difference in drop jump height before and after the downhill run was measured as well as changes in heart rate throughout the run. RESULTS: There was a significant curvilinear relationship between the delta neuromuscular efficiency and total kilometers raced (R2 = 0.53, p < 0.05), and a significant inverse relationship between delta neuromuscular efficiency and the number of races > 56 km (r = -0.50, p < 0.05). Drop jump height decreased after the downhill run, and heart rate increased during the run. CONCLUSIONS: Runners who have raced an accumulated distance of > 5,000 km show a significant dissociation in the delta neuromuscular efficiency after a downhill run, compared with less experienced runners. Although possible causes for the dissociation are discussed, further research is needed. PMID- 10798796 TI - A prospective study of exercise-associated hyponatremia in two ultradistance triathletes. AB - OBJECTIVE: To study fluid and sodium balance in two ultradistance triathletes. DESIGN: Prospective case study. SETTING: An ultradistance triathlon (3.8 km swim, 180 km cycle, 42.2 km run), and during overnight recovery. Ambient air temperature at 12:00 p.m. race day was 21 degrees C, with a relative humidity of 91%. Water temperature was 20.7 degrees C. SUBJECTS: Two female ultradistance triathletes, ages 30 and 39 years, who were participating in a larger study investigating weight and electrolyte changes in the Ironman triathlon. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Subjects were weighed and had blood drawn for serum sodium concentration, hemoglobin, hematocrit, arginine vasopressin, and aldosterone concentration prior to and after the race, and at 8:00 a.m. the following morning. Sodium and fluid intake and urinary output were measured during recovery. RESULTS: Both subjects developed mild hyponatremia (Na 131 and 130 mmol/L) during the race, with a weight gain (0.5 and 1.5 kg). Neither subject had large sodium losses (24 mmol and 20 mmol). Fluid consumption was 733 ml/h and 764 ml/h. Plasma volume increased during the race (25 and 16%). Arginine vasopressin (AVP) levels were not elevated in either subject (1.2 and 1.9 pmol/L). Both subjects demonstrated a water excess during the race (1.5 and 2.5 L), and lost weight during recovery (2.0 and 4.5 kg). CONCLUSIONS: Hyponatremia resulted from fluid retention in the extracellular space, without evidence of large sodium losses or inappropriate AVP secretion. PMID- 10798797 TI - An unusual cause of knee pain in an adolescent basketball player. PMID- 10798798 TI - Avulsion fracture of the tibial tubercle with avulsion of the patellar ligament in an adolescent female athlete. PMID- 10798799 TI - Unusual cause of knee pain mimicking meniscal pathology in a 14-year-old baseball player. PMID- 10798800 TI - External iliac artery endofibrosis in a triathlete. PMID- 10798801 TI - Use of artificial neural networks in the clinical staging of prostate cancer: implications for prostate brachytherapy. AB - PURPOSE: This review describes two studies to evaluate artificial neural networks (ANNs) in prostate cancer staging. In the first study, an ANN was trained to identify prostate cancer patients at low risk of lymph node spread (LNS). The second study evaluated an ANN to predict capsular penetration (CP) in men with clinically localized prostate cancer. An accurate assessment of lymph node status will help identify those brachytherapy patients in whom lymphadenectomy can be avoided. The accurate prediction of CP can help determine the appropriateness of brachytherapy as a treatment option. MATERIALS AND METHODS: An ANN to predict LNS was trained and tested using a database from one institution (n = 4,133) and validated using two databases (n = 330 and n = 227) from different institutions. The clinical variables used were clinical stage (cTNM), Gleason sum, and prostate specific antigen concentration (PSA). The ANN to predict CP was trained and validated with data from a single institution (n = 409). The variables used were age, race, PSA, PSA velocity, Gleason sum, and cTNM. RESULTS: The LNS ANN was able classify 76%, 75%, and 30% of the patients in each database as being at low risk of LNS with 98% accuracy. The CP ANN correctly identified CP in 25 (84%) of patients and produced 5 (16%) false-negative predictions. CONCLUSIONS: These preliminary results suggest that ANNs can be useful in staging prostate cancer. If sufficiently accurate ANNs can be developed and tested, they have the potential to increase the accuracy of clinical staging and thus improve treatment decisions. PMID- 10798802 TI - Brachytherapy in patients with small prostate glands. AB - PURPOSE: To better define the role of small prostate volume in selecting patients for brachytherapy. MATERIALS AND METHODS: Thirty patients with a transrectal ultrasound (TRUS)-based prostate volumes less than 20 cc were treated at the University of Washington by permanent isotope implantation for prostatic carcinoma. Preimplant TRUS studies were taken at 0.5-cm intervals from the base of the gland to the apex. Planning margins of 1 to 5 mm were added to the prostatic margins, and sources were routinely planned to be placed as much as 5 mm outside of the prostatic margin. The prescription dose was 144 and 115 Gy for full-dose iodine 125 and palladium 103 monotherapy, respectively. For patients receiving supplemental external-beam irradiation, the implant doses were 120 and 90 Gy for 125I and 103Pd, respectively. The morning following the implant, axial computed tomographic (CT) images of the prostate were obtained at 0.5-cm intervals with patients in the supine position. Follow-up ranged from 11 to 28 months (median 21 months). RESULTS: The median coverage of the postimplant prostate volume by the prescription dose was 92%. To calculate the incidence of source migration, the number of sources placed at the time of implant was compared with the number identified on postimplant CT scan. The median number of sources implanted was 84 (range 65-103) compared to an average of 82 identified postoperatively, which is consistent with a source migration rate of two. A median of 31 sources appeared to be outside of the prostatic margins, as identified on postimplant CT scan (range 14-53). Of the 23 patients contacted at the time of this report, one had developed acute postimplant urinary retention that resolved within 2 weeks of implantation. At last follow-up, patient pre- and postimplant AUA scores were not substantially different, with the median AUA score increasing from 7 (range 2-21) to 8 (range 1-27). CONCLUSIONS: Patients with small prostate volumes appear to have acceptable morbidity and target coverage with prostate brachytherapy. Based on the data reported here, we do not believe that a small prostate volume in itself is a contraindication to brachytherapy. PMID- 10798803 TI - Treatment of extraprostatic cancer in clinically organ-confined prostate cancer by permanent interstitial brachytherapy: is extraprostatic seed placement necessary? AB - PURPOSE: Successful treatment with ultrasound-guided transperineal interstitial permanent prostate brachytherapy (TIPPB) relies on effective radiation coverage of intraprostatic and clinically occult extraprostatic cancer. This study examines prostatectomy findings as they relate to treatment of extraprostatic extension (EPE) of cancer and TIPPB techniques and dosimetry. MATERIALS AND METHODS: A total of 313 prostatectomy specimens from patients with clinical tumor classification T1-T2b adenocarcinomas, serum prostate-specific antigen <20 ng/mL, and Gleason score <8 were whole mounted and evaluated for intraprostatic cancer volume and extraprostatic radial distance, area of perforation, and cancer density. From these data, extraprostatic cancer volume is calculated and used to estimate extraprostatic tumor control probabilities using the linear quadratic radiobiological model and Poisson statistics. TIPPB dose-gradient characteristics at the prostate periphery are examined. RESULTS: Intraprostatic cancer volume ranges from 0 to 38 cc, whereas extraprostatic cancer volume ranges from 0 to 4.6 cc (mean 0.06 cc). The radial distance of EPE ranges from 0 to 4.4 mm (mean 0.18 mm). The ratio of extraprostatic to intraprostatic cancer volume ranges from 0% to 18% (mean 0.4%). CONCLUSIONS: Only small amounts of clinically occult extraprostatic cancer were identified in the majority of specimens with EPE. Tumor control probability calculations suggest that this volume of cancer may be treated effectively with TIPPB. Treatment of this cancer possibly is achieved with an intraprostatic implant, but treatment of all cancers identified in this study suggests that some extraprostatic seed placement is desirable. PMID- 10798804 TI - Modified uniform seed loading for prostate brachytherapy: rationale, design, and evaluation. AB - PURPOSE: Despite a multitude of preplanning approaches and seed loading philosophies, multiple groups have reported comparable results in terms of biochemical disease-free survival and complication rates. None of the seed loading philosophies have been proven superior and no generally accepted loading philosophy has been adopted by the brachytherapy community. MATERIALS AND METHODS: Review of the preplanning philosophy, preplanning dosimetry, and preimplant evaluation is presented. RESULTS: A modified uniform seed loading approach is used because of its ability to deliver a relatively homogeneic dose to the prostate gland with a periprostatic margin, to routinely maintain the urethral dose at approximately 115% of the prescribed dose and to avoid significant rectal and urinary toxicity. CONCLUSION: Of the commonly utilized seed loading philosophies, a modified uniform approach delivers a relatively homogeneic dose to the prostate gland/periprostatic region and is the most forgiving of local and systematic errors in seed placement. PMID- 10798805 TI - Can the cost of permanent prostate implants be reduced? An argument for peripheral loading with higher strength seeds. AB - PURPOSE: To evaluate the economic impact of higher vs. lower strength permanent radioactive seeds used for prostate brachytherapy in the treatment of localized adenocarcinoma of the prostate. MATERIALS AND METHODS: Treatment plans for 50 patients who received an iodine 125 implant as monotherapy for favorable risk prostate cancer were reviewed and specific activity (mCi/cc) was determined for prostate volumes that ranged from 12 to 87 cc. Current prices for individual model 6711 125I seeds were obtained from the manufacturer (Nycomed/Amersham). Total seed costs for three theoretical prostate implant volumes of 25, 40, and 55 cc were calculated using seed strengths of 0.25, 0.34, 0.50, and 0.75 mCi/seed. RESULTS: Specific activities for prostate volumes of 25, 40, and 55 cc were 1.25, 1.15, and 1.00 mCi/cc, respectively. Total seed cost was inversely related to seed strength. For a 25-cc prostate the cost ranged from $1,890 (0.75 mCi/seed) to $5,625 (0.25 mCi/seed), for a 40-cc prostate $2,745 to $8,280, and for a 55-cc prostate $3,285 to $9,900. For a medium-sized gland (40 cc), the treatment plan using a seed strength of 0.75 mCi/seed resulted in a total seed cost of $2,745 vs. $6,075 for a plan using an activity of 0.34 mCi/seed. This savings of approximately 55% in total seed cost between seed strengths of 0.75 and 0.34 mCi/seed held true for small (25 cc) and large (55 cc) prostate volumes as well. CONCLUSIONS: Given the current prices for 125I individual seeds, prostate implants using a peripheral loading technique with higher activity seeds may result in a significantly lower material cost than techniques using lower activity seeds. However, issues regarding morbidity as well as sensitivity to source placement error need to be addressed further before any final conclusions can be made. PMID- 10798806 TI - Three-dimensional real-time magnetic resonance-guided interstitial prostate brachytherapy optimizes radiation dose distribution resulting in a favorable acute side effect profile in patients with clinically localized prostate cancer. AB - PURPOSE: A large number of men are diagnosed with early-stage prostate cancer as a result of prostate-specific antigen (PSA) screening. For many of these men, prostatectomy results in long-term freedom from biochemical and clinical failure. Despite limited follow-up data, ultrasound-guided prostate brachytherapy has gained acceptance as a treatment for early-stage prostate cancer, in part due to its favorable side effect profile and brief recovery period. We report on the use of three-dimensional real-time magnetic resonance (MR) guidance, which has several advantages compared with biplanar ultrasound guidance for prostate brachytherapy, including enhanced geometric and dosimetric feedback during the procedure. MATERIALS AND METHODS: Median clinical target volume coverage of 96% was achieved using MR guidance. The ability to define more precisely the clinical target volume with MR and the use of real-time assessment of dose distribution resulted in an excellent acute side effect profile. Only 19% of patients required oral alpha1 blockers for postimplant urethritis and 9% required temporary reinsertion of the Foley catheter due to acute urinary obstruction. CONCLUSIONS: These results compare favorably to those of ultrasound-guided brachytherapy. Further follow-up is needed to ascertain the impact this technique will have on cancer control and long-term quality of life. PMID- 10798807 TI - Intraoperative dosimetric representation of the real-time ultrasound-guided prostate implant. AB - PURPOSE: To describe a method of creating an intraoperative dosimetric representation of the real-time ultrasound-guided prostate implant. MATERIALS AND METHODS: An intraoperative dosimetry system (Multi Media Systems [MMS]) captures transverse ultrasound images after peripheral needles have been implanted in the prostate. The prostate contour and needle positions are outlined on the system. The volume of the prostate with needles in place is calculated. As seeds are deposited in the actual implant, the positions of the seeds are marked on the intraoperative system. Following implantation of the peripheral needles, the resulting isodose lines are displayed. The interior needles are inserted into the prostate, and these positions are captured on the system. As seeds are deposited through these needles into the prostate, their positions are captured on the planning system. When the implant is complete, the final dose coverage and dose volume histogram can be visualized. RESULTS: Ten consecutive patients underwent iodine 125 implants using real-time intraoperative isodose generation. The ratio of the preneedle prostate volume to postneedle prostate volume ranged from 0.89 to 1.0 (median 0.97). The calculated dose delivered to 90% of the prostate volume from the dose volume histogram (D90) ranged from 146.5 to 194 Gy (median 174.75 Gy). The percentage of the prostate covered by 240 Gy ranged from 14.6% to 59% (median 40.75%). CONCLUSION: Dosimetric representation of the real-time ultrasound-guided prostate implant can be achieved and demonstrates the efficacy of this brachytherapy technique. PMID- 10798808 TI - A nomogram to compensate for intraoperative prostate edema during transperineal brachytherapy. AB - PURPOSE: Prostate edema during and following prostate brachytherapy may have a negative impact on implant dosimetry. The purpose of this study was to assess the magnitude of prostate edema during the operative procedure and to develop a nomogram of isotope activity required for compensation of such intraoperative prostate volume changes. MATERIALS AND METHODS: Twenty-five consecutive patients with early-stage, localized adenocarcinoma of the prostate underwent ultrasound guided transperineal interstitial permanent prostate brachytherapy with either iodine 125 or palladium 103. Transrectal ultrasound volume studies of the prostate were performed before and during the implant procedure. Computed tomography-based postimplant dosimetry was performed 3-4 weeks after surgery. RESULTS: A median intraoperative prostate volume increase after insertion of applicator needles of 10.4% (range 1.2-32.5%) was identified. A correlation of 0.55 (95% confidence interval -0.78 to -0.19) between the minimum dose covering 90% of the prostate volume (%D90) and the amount of edema was identified. An algorithm and nomogram was developed to calculate the extra isotope activity necessary to compensate for intraoperative edema. CONCLUSIONS: Prostate edema occurs at the time of transperineal needle placement. A negative correlation was found between the amount of edema and dose coverage of the prostate (%D90). Therefore, to cover the prostate volume adequately, additional isotope activity is required when preoperative treatment planning is performed. This nomogram can be used to compensate for such volume changes. PMID- 10798809 TI - Is there a role for antibiotic prophylaxis in transperineal interstitial permanent prostate brachytherapy? AB - PURPOSE: There are few data to guide the physician on the use of prophylactic antibiotic(s) for prostate brachytherapy. The purpose of this study was to evaluate the symptomatic urinary tract infection (UTI) rate after performing transperineal interstitial permanent prostate brachytherapy (TIPPB) in conjunction with cystoscopy. MATERIALS AND METHODS: One-hundred twenty-five patients underwent TIPPB and cystoscopy. All patients received intravenous perioperative antibiotic prophylaxis. No postimplant antibiotic medication was prescribed. All patients were evaluated at 1-month follow- up for symptomatic UTI. No screening (U/A, C+S) was performed for asymptomatic patients. Any UTI within 1 month of TIPPB was considered a complication and scored as an infection. RESULTS: Of 125 patients who underwent TIPPB and cystoscopy, one patient (1%) developed a symptomatic UTI. In our study, a one-time perioperative intravenous dose of cefazolin (Ancef) without additional postoperative antibiotics resulted in an overall symptomatic UTI rate of 1%. Hence, additional postoperative antibiotics may not be warranted, thus providing a cost saving (500 mg of ciprofloxacin orally, two times a day for 5 days at a cost of $44.95) and reducing the potential risk of antibiotic resistance. CONCLUSIONS: When cystoscopy is used in conjunction with TIPPB, perioperative antibiotic prophylaxis is recommended. However, due to the low infection rate expected from TIPPB, postimplant antibiotic use is not recommended. As a result of the low infection rate anticipated from TIPPB and cystoscopy, a large multiinstitutional trial is needed to determine the necessity of antibiotic prophylaxis for TIPPB and cystoscopy. PMID- 10798810 TI - Cystourethroscopic findings before and after prostate brachytherapy. AB - PURPOSE: To assess the role of cystourethroscopy in predicting the risk of postimplant urinary retention. MATERIALS AND METHODS: Fifteen unselected prostate brachytherapy patients implanted with 125I or 103Pd under spinal or general anesthesia were studied. Following induction of anesthesia, the patient was placed in the lithotomy position and cystourethroscopy performed using a 17 sheath and a 30 degrees lens. Irrigation pressure was 100-cm water. A photograph was taken from the level of the verumontanum. At completion of the implant procedure, a second cystourethroscopy was performed and another photograph taken. The degree of obstruction was rated using a 3-point scale. Each patient was contacted at the time of this report to update postimplant morbidity information with follow-up from 1 to 10 months. RESULTS: The patients' preimplant cystourethroscopic findings ranged from minimal to complete occlusion. There was no clear relationship between the American Urologic Association (AUA) score or preimplant prostate volume and the degree of obstruction. Nearly all patients had some increased physical obstruction following completion of the procedure, but only 8 of 14 patients had an increase in obstruction grade. Six patients were completely obstructed at the completion of the implant procedure, only one of whom developed urinary retention. CONCLUSIONS: Marked variability in cystourethroscopy findings do not appear to have a strong influence on the likelihood of postimplant urinary retention and are not a reliable predictor of retention. PMID- 10798811 TI - Dynamic cystography can replace cystoscopy following prostate seed implantation. AB - PURPOSE: To determine if dynamic cystography can replace cystoscopy following prostate seed implantation. MATERIALS AND METHODS: Three hundred three consecutive patients underwent prostate seed implantation using the real-time method. Following implantation, 60 cc of contrast media was injected into the bladder using continuous fluoroscopy. If any seed motion was detected, cystoscopy was performed and any seeds encountered were removed. Patients were followed after discharge for any seeds passed in the urine. RESULTS: Of the 303 patients, 3 (1%) were found to have one or more seeds in the bladder. No patients passed seeds following discharge. There were no adverse events associated with the cystography. CONCLUSION: Dynamic cystography can replace cystoscopy as the postimplant study to evaluated misplaced intravesical or urethral seeds. PMID- 10798812 TI - Influence of prophylactic dexamethasone on edema following prostate brachytherapy. AB - PURPOSE: This work evaluates the prophylactic administration of dexamethasone as an adjunct to prostate brachytherapy to determine the effect of dexamethasone on postimplant edema, dosimetric quality, catheter dependency, and clinical outcome as assessed by serial American Urological Association (AUA) symptom score determinations. MATERIALS AND METHODS: In a controlled study, 22 unselected patients were placed alternately into either a dexamethasone or control arm. RESULTS: There was no statistically significant difference in volume between the experimental and control groups on either day 0 or day 28. Accordingly, the day 0 dosimetric parameters, short-term catheter dependency, and serial AUA symptom score determinations were equivalent in both patient groups. However, the patients receiving dexamethasone had a markedly smaller volume on day 3, but 9 of 11 experienced rebound edema with a volume on day 14 significantly greater than day 3. CONCLUSIONS: Because of the small number of patients evaluated, additional studies are essential to confirm these findings. At the present time, we do not recommend this dexamethasone schedule as a routine component of prostate brachytherapy. PMID- 10798813 TI - Prior transurethral resection does not increase morbidity following real-time ultrasound-guided prostate seed implantation. AB - PURPOSE: Patients with localized prostate cancer who had a prior open prostatectomy or transurethral resection of the prostate (TURP) for benign prostatic hyperplasia (BPH) may be at risk for greater morbidity when treated with brachytherapy. This analysis examines the morbidity following brachytherapy using the real-time method to determine if patients with a history of TURP are at increase risk for developing complications. MATERIALS AND METHODS: An ultrasound guided transperineal interactive prostate seed was implanted in 419 patients with T1-T2 prostate cancer. All patients were implanted using a peripheral weighting of sources (75%) with the interior sources placed at least 5 mm from the urethra. The patients were divided into two groups: group 1 consisted of 376 patients (89.7%) without a prior TURP, and group 2 consisted of 43 patients (10.3%) who had a TURP prior to their implant. The mean age, prostate-specific antigen level, Gleason score, clinical stage, prostate volume, isotope implanted, and number of patients treated with neoadjuvant hormone therapy were comparable for both groups. RESULTS: Median follow-up for group 1 was 12 months and for group 2 was 18 months. No patients suffered from radiation-related proctitis or cystitis in either group of patients. Two patients in group 2 implanted with iodine 125 and who had a history of two prior TURPs developed mild superficial urethral necrosis (SUN). The actuarial freedom from developing superficial urethral necrosis at 4 years was 84% in patients with a history of prior TURP. There were no episodes of SUN in group 1 and no cases of incontinence reported in either group of patients. The actuarial rate of potency was 78% at 2 years. CONCLUSION: Whereas other techniques of seed implantation report incontinence in patients who had a prior open prostatectomy or TURP, the real-time method combined with peripheral loading avoids this complication. PMID- 10798814 TI - Evolution of techniques for ultrasound-guided palladium 103 brachytherapy in 950 patients with prostate cancer. AB - PURPOSE: Brachytherapy is an effective, low-morbidity, cost-effective treatment for adenocarcinoma of the prostate. Although both iodine 125 and palladium 103 have been used extensively over the last 14 years, 103Pd (Theraseed) provides a more rapid initial dose and therefore better control of higher Gleason grade tumors. We assessed a highly refined methodology for 103Pd brachytherapy in patients with prostate cancer and compared outcomes before and after making adjustments in technique. MATERIALS AND METHODS: Standard brachytherapy techniques were modified to include combination of preplanning with real-time adjustment, placing all needles at once to minimize prostate movement, using monitored anesthesia control sedation during preoperative volume study and cystoscopy, and several other small but important changes. Charts were reviewed from 950 patients treated with 103Pd implants from 1991 through 1999. Prostate specific antigen (PSA) and biopsy data before and after 1993 were compared for 733 patients whose data qualified for analysis. RESULTS: At 1 and 5 years after treatment, 85-88% of patients had stable PSA <1.5 ng/mL. Biopsies were negative in 90-94% at 2 years. CONCLUSION: The modification in techniques provides positive results that remain stable over time. PMID- 10798815 TI - Use of conformal high-dose rate brachytherapy for management of patients with prostate cancer: optimizing dose escalation. AB - PURPOSE: We reviewed our institution's experience treating patients with locally advanced prostate cancer using high-dose rate (HDR) conformal brachytherapy. Treatment technique, interim results, and toxicity are reviewed. MATERIALS AND METHODS: From November 1991 to May 1998, 161 patients with locally advanced prostate cancer were treated on a dose escalation trial of external-beam radiation therapy (EBRT) combined with HDR conformal brachytherapy boosts. Patients with any of the following characteristics were acceptable for enrollment: pretreatment prostate-specific antigen (PSA) > or =10 ng/mL, Gleason score > or =7, or clinical stage T2b or higher. All patients received pelvic EBRT to a median dose of 46 Gy. Transperineal ultrasound-guided temporary HDR brachytherapy implant boosts were performed in the first, second, and third week of EBRT. Seventy-two patients were boosted with three implants (boost dose 5.50 to 6.50 Gy), and 89 patients received two implants (boost dose 8.25 to 10.50 Gy). All implants were placed using interactive real-time dosimetry. Biochemical failure was calculated using the American Society for Therapeutic Radiology and Oncology Consensus Panel definition. Median follow-up was 2.8 years (range 0.3 7.2). RESULTS: The 2- and 5-year actuarial biochemical control rates were 86% and 67%, respectively. The 5-year actuarial biochemical control rates for patients with a pretreatment PSA < or =3.9, 4.0-9.9, 10.0-19.9, and > or =20.0 ng/mL were 80%, 87%, 56%, and 54%, respectively. Factors associated with biochemical failure on multivariate analysis included the pretreatment PSA, Gleason score, PSA nadir, and time to PSA nadir. A total of 6 patients (4%) developed grade 3 late toxicity consisting of urethral stricture (5 patients) or incontinence (1 patient). Forty- four patients (27%) developed impotence after radiation therapy. CONCLUSIONS: Conformal HDR brachytherapy boosts appear to offer a safe, reproducible, and effective method of dose escalation in patients with locally advanced prostate cancer treated with RT. Interim results with this technology reveal biochemical control rates paralleling those achieved with three-dimensional conformal EBRT and other forms of treatment. PMID- 10798817 TI - Use of image coregistration in salvage prostate brachytherapy. AB - PURPOSE: We describe a method of performing salvage prostate brachytherapy on patients whose initial implant was suboptimal. This technique uses an image correlation algorithm only previously used to fuse postimplant magnetic resonance and computed tomographic (CT) images. Here, the initial postimplant CT and the second preimplant volume study are coregistered to plan delivery of the salvage implant. MATERIALS AND METHODS: Two early-stage patients had salvage implants performed with this technique, in which only a limited number of sources were visible on the ultrasound images. The dosimetric results of the first implant were displayed on the preplan generated for the second procedure. The planned total dose then was visualized prior to salvage implant. RESULTS: The implants were performed without complication. Rectum and urethra doses remained acceptable. In each case, the improvement in coverage of the gland was dramatic (V80 coverage improved from 65.2% and 47.3% to 93.1% and 92.2%, respectively), precluding the need for further intervention. CONCLUSIONS: Coregistration of the postimplant CT scan to an ultrasound volume study can be quantifiably and reliably performed. The resulting image set can be used to guide needle placement during a second salvage implant to achieve much improved dosimetric coverage of the gland. PMID- 10798816 TI - Use of radioimmunoscintigraphy with indium-111-labeled CYT-356 (ProstaScint) scan for evaluation of patients for salvage brachytherapy. AB - PURPOSE: Indium 111 capromab pendetide radioimmunoscintigraphy (ProstaScint) has been used to detect occult recurrent carcinoma after radical prostatectomy. MATERIALS AND METHODS: We evaluated the role of ProstaScint in 24 men with rising prostate-specific antigen (PSA) level following definitive radiation therapy to differentiate between local and distant recurrence in this patient population. RESULTS: ProstaScint scan detected prostatic uptake only in 16 patients, extraprostatic uptake in 5, both prostatic and extraprostatic uptake in 1, and no uptake in 2. Ten of 21 patients with prostatic uptake had positive biopsies, 5 had negative biopsies, and 6 were not biopsied. Two of the three patients with negative scans had positive biopsies, and the third patient was not biopsied. Three patients had evidence of osseous metastasis on radionuclide bone scan, two corresponding to the sites detected on ProstaScint. All three patients had abnormal uptake beyond the prostatic fossa with (n =2) or without (n = 1) prostatic uptake. There were no positive bone scans in patients without extraprostatic uptake on ProstaScint. CONCLUSIONS: ProstaScint scan is useful in detecting occult recurrence outside the prostate in patients with rising PSA following radiation therapy. Compared to data from radical prostatectomy, ProstaScint scans in these patients reveal a higher prevalence of abnormal uptake in the prostate and less frequent extraprostatic uptake. PMID- 10798818 TI - Techniques to achieve optimal seed placement in salvage and primary brachytherapy for prostate cancer. AB - PURPOSE: To delineate flexible, interactive prostate brachytherapy techniques relying extensively on intraoperative biplane ultrasound and fluoroscopic visualization as developed over a period of 10 years in a brachytherapy program involving more than 2,500 patients treated to date. MATERIALS AND METHODS: Specialized techniques were developed and refined in pretreatment transrectal ultrasound evaluation, interactive seed placement, and postoperative evaluation of implant quality. Special methods were also designed and implemented for treatment of salvage patients and otherwise difficult cases involving large prostates, interfering pubic bone structure, and prostatic defects due to prior transurethral resection of the prostate. RESULTS: Favorable clinical results, including 79% actuarial disease-free survival at 5 years with low morbidity, have been recently reported in 543 patients with localized primary prostate cancer treated by brachytherapy using the specialized techniques. Promising results have also been recently described in 49 patients who had failed prior radiotherapy and then underwent salvage brachytherapy. Among the techniques found useful were the judicious use of curved stylets and disposable and nondisposable needles, as well as monitoring and minimizing intraoperative prostate movement and compensating for unavoidable movement. CONCLUSIONS: Prostate brachytherapy has attracted renewed interest, largely due to advancements in technique. The presented techniques allow brachytherapy to be performed optimally in a broad range of patients. PMID- 10798819 TI - The prognostic value of thought disorder in psychotic depression. AB - Thirty-eight patients with psychotic depression were compared to 40 patients with severe, nonpsychotic depression. Psychiatric variables, along with demographic data, were used to predict relapse over a 7-year period in a multiple regression model. Initial formal thought disorder was a strong predictor of relapse during the follow-up (F = 43, p < .001). The predictive value of this symptom in psychotic depression was not enhanced by the addition of other symptomatic or demographic variables. It appears that individuals with psychotic depression who experienced formal thought disorder are much more likely to have a relapse within a 7-year period than other people with depression. PMID- 10798820 TI - Lamotrigine compared with lithium in mania: a double-blind randomized controlled trial. AB - BACKGROUND: Preliminary data from case reports and small open trials suggest a role for lamotrigine in the treatment of bipolar disorder, although controlled data for the manic phase are lacking. METHOD: Thirty inpatients with a DSM-IV diagnosis of bipolar I disorder, currently manic, were randomly allocated to receive either lamotrigine (25 mg once daily for 1 week, 50 mg once daily for the second week, and 100 mg once daily for the last 2 weeks) or lithium (400 mg twice daily) in a 4-week randomized, double-blind, clinical trial. RESULTS: Both treatments improved symptoms of mania, as assessed by the Mania Rating Scale, Brief Psychiatric Rating Scale, Clinical Global Impression severity and improvement scales, and the Global Assessment of Functioning scale. There were no significant differences between groups at any time point, suggesting that the dose escalation required for lamotrigine did not adversely affect its onset of action. Secondary outcome measures, including the use of lorazepam as rescue medication, did not differ between the groups. No significant adverse events were noted in either group. CONCLUSION: In this pilot study, lamotrigine was as effective as lithium in the treatment of patients with bipolar disorder hospitalised for acute mania. PMID- 10798821 TI - Efficacy and tolerability of olanzapine in elderly patients with psychotic disorders: a prospective study. AB - Olanzapine is a novel antipsychotic effective in reducing positive and negative symptoms of schizophrenia and with a safe side-effect profile. Premarketing trials, however, included only a few elderly patients. Further data are needed regarding the effects of olanzapine in the elderly and those with comorbid medical illness. In this pilot study, 11 hospitalized patients (age range 60-85 years) who manifested symptoms of psychosis related to schizophrenia and schizoaffective disorders were treated with olanzapine (dose range, 5-20 mg/day). Efficacy and safety were assessed by the Positive and Negative Syndrome Scale (PANSS), Clinical Global Impression Scale (CGI), Extrapyramidal Symptom Rating Scale (ESRS), Mini-Mental State Examination (MMSE), Calgary Depression Scale For Schizophrenia (CDSS), EKG, physical examination, and various laboratory tests. Seven patients responded to treatment and all of them showed improvement in both positive and negative symptoms, with greater reduction in positive symptoms. Treatment was discontinued in 2 patients whose symptoms showed no improvement or worsened. The CGI showed significant improvement in 9 patients, remained the same in 1, and worsened in 1 patient. ESRS showed significant reduction from baseline to final visit. Of the 10 patients who cooperated for MMSE, 9 had improved scores. The CDSS showed significant reduction in scores from baseline to final visit. No significant changes were noted in laboratory tests, prolactin levels, EKG, and physical examination. Concomitant administration of lorazepam, carbamazepine, divalproex sodium, and lithium carbonate caused no adverse consequences. The reduction of positive and negative symptoms, lack of significant extrapyramidal symptoms and other side effects, and lack of any significant drug interaction suggest that olanzapine may be a safe and effective antipsychotic medication in the elderly. PMID- 10798822 TI - The psychological and physical health of hospice caregivers. AB - This study explores the psychological distress of caring for a dying family member and examines the differences in depression, anxiety, health, social and occupational functioning, and social support among hospice caregivers and community controls. It compares psychological functioning of spousal and adult child hospice caregivers. Caregivers of terminally ill hospice patients were assessed prior to death as a part of a longitudinal bereavement study. Caregivers reported experiencing higher levels of depression, anxiety, anger, and health problems than controls. Hospice caregiving was associated with deterioration in physical health and in social and occupational functioning. The comparisons between adult children and spouse caregivers revealed that levels of psychological and physical morbidity were very similar for the two generations of caregivers. An awareness of distress symptoms among hospice caregivers could lead to timely proactive clinical intervention that may prevent bereavement complications. PMID- 10798823 TI - Bipolar II depression with melancholic features. AB - Bipolar II depression with melancholic features has been understudied. The aims of the present study were to find the prevalence of melancholic features in bipolar II depression and in unipolar depression, and to compare melancholic with nonmelancholic bipolar II/ unipolar depression in private practice. One hundred and sixty two consecutive unipolar and bipolar II depressed outpatients were interviewed with the DSM-IV Structured Clinical Interview, the Montgomery-Asberg Depression Rating Scale, and the Global Assessment of Functioning Scale. Melancholic features were present in 19.2% of bipolar II patients and in 22.6% of unipolar patients, a nonsignificant difference. Melancholic bipolar II patients versus nonmelancholic bipolar II patients had significantly more psychosis and higher severity. All the other variables (age, age at onset, gender, illness duration, recurrences, atypical features, chronicity, comorbidity) were not significantly different. Melancholic bipolar II patients versus melancholic unipolar patients were not significantly different. Nonmelancholic bipolar II patients versus nonmelancholic unipolar patients had significantly lower age, lower age at onset, more atypical features, and more comorbidity. The prevalence of melancholic features in bipolar II depression in private practice was higher than previously reported in academic centers. PMID- 10798824 TI - Hair loss in psychopharmacology. AB - Medication-induced alopecia is an occasional side effect of many psychopharmaceuticals. Most of the mood stabilizer and antidepressant drugs can lead to this condition. Some antipsychotic and antianxiety agents induce alopecia. Hair loss is also related to hypothyroidism, which can be induced by lithium and other agents. Alopecia might not be reported by some people, but physicians should be aware of this potential problem which may contribute to noncompliance. Lithium causes hair loss in 12-19% of long-term users. Valproic acid and/ or divalproex precipitates alopecia in up to 12% of patients in a dose dependent relationship. Incidences up to 28% are observed with high valproate concentration exposures. These pharmaceuticals also can change hair color and structure. The occurrence of carbamazepine-induced alopecia is at or below 6%. Hair loss is less common with other mood stabilizers. Tricyclic antidepressants, maprotilene, trazodone, and virtually all the new generation of antidepressants may on rare occasions lead to alopecia. The same applies to haloperidol, olanzepine, risperidone, clonazepam, and buspirone, but not to other neuroleptics, benzodiazepines, or barbiturates, selected antihistamines, and antiparkinsonians. Discontinuation of the medication or dose reduction almost always leads to complete hair regrowth. The therapeutic value of mineral supplements remains unclear. PMID- 10798825 TI - Risk of suicidality in depression with serotonergic antidepressants. AB - Since depression is a risk factor for suicidal thoughts and behaviors, and since suicidal behaviors are associated with low serotonin activity, are selective serotonin reuptake inhibitors (SSRIs) more effective than other antidepressants in treating suicidality in depressed patients? There is inconclusive evidence for and against this hypothesis. However, all studies suggest that antidepressants are effective treatments of suicidal ideations and behaviors, and SSRIs have been shown to have prophylactic effects in preventing suicidal behaviors. Although some reports suggest that SSRIs might increase suicidal ideations and behaviors, the results of large, double-blind studies do not suggest a causal relationship between pharmacotherapy and the emergence of suicidality. Undertreatment of depression and therapeutic failure are more significant problems with the use of antidepressants in suicidal patients than the risk of using antidepressants in overdose. Prescribing inadequate doses of antidepressants is therefore a source of overlooked risk. PMID- 10798826 TI - Neuroleptic malignant syndrome and severe thrombocytopenia: case report and literature review. AB - We report an unusual case of thrombocytopenia associated with neuroleptic malignant syndrome (NMS). A 31-year-old Black male with a history of hypertension, partial seizures, and schizophrenia developed acute rigidity closely followed by severe hyperpyrexia (temperature 102 degree F), tachypnea, and tachycardia. His home medications at the time of presentation included propanolol 10 mg tid, haloperidol 10 mg bid, sodium valproate 500 mg bid, benztropine 1 mg bid, and haloperidol decanoate 100 mg i.m. every 3 weeks, from another psychiatric facility. Despite vigorous therapy for the hyperthermia, he rapidly developed significant hypoxia requiring mechanical ventilation. A diagnosis of neuroleptic malignant syndrome was made and the patient continued to receive aggressive supportive care. On hospital day 2 his platelet count dropped to 47,000/microl and bottomed out at 36,000/microl by day 3 with other blood cell counts remaining within normal limits. Over the next few days he showed rapid clinical improvement with normalization of his blood chemistries and he was discharged home after 5 days of hospitalization in good condition. PMID- 10798827 TI - Are stimulants overprescribed for youths with ADHD? AB - Critics of stimulant treatment for youths with attention deficit hyperactivity disorder (ADHD) have increased their rhetoric of late, contending that the leading medication for it, Ritalin, is vastly overprescribed. Additionally, they claim that Ritalin (methylphenidate) is inherently dangerous and that the entire system of the diagnosis and treatment of ADHD is seriously flawed. The critics view the underlying reason for the "epidemic" as societal, due to our modern pace of living, our competitive society, and our consumer emphasis. Rejoinders to and clarifications of the more tangible points of the critics are presented, followed by a discussion of some more practical and legitimate concerns for researchers in this area. These concerns include changes within the ADHD category, the clinical need for multiple sources of diagnostic data, infrequent teacher-physician communication, problematic ADHD/conduct disorder comorbidity in adolescence, and the limited amount of community-based research. PMID- 10798828 TI - Are vestibular hair cells excited to death by aminoglycoside antibiotics? AB - Aminoglycoside antibiotics are commonly prescribed throughout the world for the treatment of serious gram-negative bacterial infections. However, their use is limited by nephrotoxicity and ototoxicity. Although the traditional explanation for their ototoxicity in cochlear and vestibular hair cells has been the inhibition of mitochondrial protein synthesis, evidence is accumulating to suggest that many aminoglycosides cause excitotoxicity in hair cells as a result of their agonist action at the polyamine site on the N- methyl-D-aspartate (NMDA) receptor. The aim of this minireview is to summarise and critically evaluate this evidence and to consider the therapeutic implications of this hypothesis. PMID- 10798829 TI - Vestibular disorders and impaired path integration along a linear trajectory. AB - The goal of this study was to determine if people use vestibular information to keep track of their positions while walking through a simple course. Subjects were normals and patients with chronic peripheral vestibulopathies-- each of whom were tested once--and patients with acoustic neuromas tested pre-operatively and one and three weeks post-operatively. Subjects walked over a straight course, 7.62 m, with their eyes open and then with their eyes closed. The time needed for task performance, the forward distance subjects walked before veering, and the lateral distance subjects veered from the straight ahead were recorded. The angle of veering was then calculated. Normals were able to perform this task easily with eyes open or closed. With eyes closed pre-operative acoustic neuroma subjects walked significantly shorter distances before veering than normals but did not veer significantly more than normals or take longer than normals to perform the task. Chronic vestibulopathy subjects, by contrast, were significantly impaired compared to normals on all measures. With eyes open within a week after acoustic neuroma resection subjects could perform the task as well as normals. With eyes closed, however, post-operative subjects were impaired compared to their own pre- operative levels, but they had returned to their pre operative levels at the second post-operative test. Ataxia was only weakly correlated to any measures and tumor size was not related to performance. These findings support the hypothesis that vestibular input is used for spatial orientation during active motion. PMID- 10798830 TI - Short-term vestibular responses to repeated rotations. AB - To investigate the short-term vestibular habituation, we performed the 4 successive velocity step tests on 28 volunteers, the peak velocity of which was 100 deg/sec with acceleration and deceleration of 100 deg/sec2. As the repeated rotations might alter the vestibulo-ocular reflex (VOR), sinusoidal rotations at a frequency of 0.16 Hz were also given before and after the 4 successive velocity step tests to investigate the changes of gain and phase of VOR. The 28 volunteers were divided into two groups, group I and II, according to their responses to repeated rotations. In group I (25 subjects, mean age 23.7 year), the mean values of the slow cumulative eye position (SCEP), the time integral of eye velocity during nystagmus, was reduced after each trial of the 4-successive velocity step tests as follows; 403.4 +/- 29 degree (1st trial), 346.2 +/- 37 degree (2nd trial), 278.3 +/- 33 degree (3rd trial) and 256.6 +/- 36 degree (4th trial). The time constant of the nystagmus was also reduced as follows; 12.9 +/- 0.78 second (1st trial), 12 +/- 0.63 second (2nd trial), 9.7 +/- 0.78 second (3rd trial) and 9.9 +/- 0.54 second (4th trial). In group II (3 subjects, mean age 28.3 year), the mean values of SCEP gradually increased; 774.3 +/- 135 degree (1st trial), 1127 +/- 178 degree (2nd trial), 1096.3 +/- 123.4 degree (3rd trial) and 1225.7 +/- 199.7 degree (4th trial). The time constant of the nystagmus increased; 15.7 +/- 2.7 second (1st trial), 22 +/- 4.5 second (2nd trial), 22.3 +/- 3.3 second (3rd trial) and 23.3 +/- 5.7 second (4th trial). The gain of vestibulo-ocular reflex (VOR) induced by sinusoidal rotations at a frequency 0.16 Hz increased in both groups: 0.59 +/- 0.03 to 0.78 +/- 0.06 (group I) and 0.65 +/- 0.07 to 1.15 +/- 0.06 (group II). Phase changes were also observed. In group I, the phase was shifted from 0.6 +/- 0.6 degree to -0.40 +/- 0.6 degree. In group II, the phase was shifted from 3.67 +/- 1.86 degree to -0.33 +/- 0.33 degree. The repeated rotation did not induce a common nystagmic response in all subjects. Thus, person to person variations should be considered in short term vestibular habituation. PMID- 10798831 TI - Measurement of vestibular ocular reflex (VOR) time constants with a caloric step stimulus. AB - A protocol is described for measuring responses to a broad-band (1-2 Hz) caloric step stimulus from which the vestibular ocular reflex (VOR) and adaptation time constants can be estimated. This novel stimulation is the caloric equivalent to a rotatory step of head acceleration. In this protocol, the ear is irrigated continuously for 5 min with water at a constant temperature. During the initial 2 min period of irrigation the subject is seated and leaning forward in a nonstimulable position (horizontal canals in a horizontal plane). This irrigation phase establishes a steady-state thermal gradient across the horizontal canal, effectively eliminating thermal dynamic properties of the caloric transmission as a confounding factor. At the end of this phase, the subject is rapidly reclined to a stimulable position (horizontal canals in vertical plane) that elicits the VOR nystagmus response to an on-step of force on the cupula. Consistent with adaptation processes, the VOR response first increases and then declines gradually over the 2-min period that the step of force is maintained. Four minutes after the onset of irrigation, the subject is rapidly returned to the nonstimulable position (off-step), which is then maintained for a final 1 min. The response after the off-step, which releases the force on the cupula, reveals reversed after-nystagmus due to adaptation. Five subjects provided caloric step responses for 26 caloric temperature conditions spanning the range from 28.4 to 43.0 degrees C. The resulting responses were fitted with an adaptation model similar to models applied to rotatory acceleration step responses. Estimates of the model parameters for robust caloric stimulation, including time constants for the VOR (18.3 sec) and for vestibular adaptation (153.2 sec), are considered in relation to corresponding values reported in the literature for rotatory and caloric vestibular stimulation. The results suggest that caloric step stimulation can be used successfully to probe VOR dynamics. PMID- 10798832 TI - Rhythmical eye-head-torso rotation alters fore-aft head stabilization during treadmill locomotion in humans. AB - A repetitive manoeuvre called torso rotation (TR) is known to temporarily reduce the gain of the horizontal vestibulo-ocular reflex by 10-15% in healthy humans. TR consists of a series of rhythmical rotations of the eyes, head and upper body executed continuously for 30 minutes while standing. Our aim was to investigate whether TR affects the ability to hold the head in a fixed fore-aft position relative to space while walking on a treadmill with eyes closed. Ten healthy subjects stood in a carefully standardized position on a stationary treadmill. The treadmill started unexpectedly and ran for 4 s at 29 cm/s. The test stimulus was a linear acceleration in the fore-aft direction at the moment of treadmill start-up. Linear head position (i.e., ability to stabilize the head) was measured during and following the stimulus. A mechanical system prevented head rotation. Two series of 60 trials were performed before TR (control 1 and control 2 series) and one after TR. Before TR, subjects drifted rearward at an average drift velocity +/- S.D. = 3.1 +/- 0.9 cm/s. This drift was reasonably stable over time within and between the two control series. After TR, head holding ability was further impaired, with subjects having more difficulty to stabilize their head after treadmill start-up. In the first 10 trials after the arrest of TR, the average drift velocity was significantly larger than before TR(6.1 +/- 1.5 cm/sec, p < 0.01). Recovery to control values followed a roughly exponential time course, with 67% recovery occurring in the first 3.4 minutes after TR. Our results indicate that TR impairs the ability to sense and/or respond to fore-aft linear accelerations of the head following treadmill start-up in the absence of vision. PMID- 10798833 TI - The influence of light on modulation of the human vestibulo-ocular reflex. AB - The goal of this study was to investigate the influence of light, without retinal slip information, on the ability to generate eye movements to compensate for head rotations. Subjects were rotated sinusoidally at 1.0, 2.0 or 3.0 Hz at a peak velocity of 30 deg/sec while they: 1) performed mental arithmetic in darkness; 2) attempted to view the remembered location of a stationary target in darkness; 3) attempted to view the remembered location of the stationary target through translucent contact lenses that allowed the passage of light but did not provide any target information (ganzfeld stimulus); 4) directly viewed the illuminated stationary target. The gain of compensatory eye movements was least while subjects viewed through the translucent contact lenses (median = 0.76), intermediate while subjects either performed mental arithmetic in darkness (median = 0.84) or attempted to view the remembered location of the target in darkness (median = 0.84), and greatest if they actually viewed the target (median = 0.95). Our findings suggest that factors other than light alone account for the increased gain of compensatory eye movements that occurs when subjects view rather than imagine a stationary target. PMID- 10798834 TI - Projective techniques and the detection of child sexual abuse. PMID- 10798835 TI - A required mind-set for child protection practice: comments on Munro (1999) PMID- 10798836 TI - Social support and the relationship between family and community violence exposure and psychopathology among high risk adolescents. AB - OBJECTIVE: The objective of this study was to examine the protective effect of social support in the relationship between exposure to violence and psychopathology. Exposure to violence in the family and exposure to violence in the community were examined separately. Exposure to violence was further divided according to whether violence was experienced as a victim or as a witness. Internalizing and externalizing forms of psychopathology, as well as post traumatic stress symptomatology were examined. METHOD: Participants consisted of 65 high-risk adolescents admitted consecutively to psychiatric inpatient units. Data were collected by means of individual interviews, self-report questionnaires, and hospital charts. RESULTS: Social support emerged as a protective factor with respect to the maladaptive effects of family violence, experienced as either a victim or as a witness. In contrast, social support did not appear to buffer the maladaptive effects of community violence, regardless of whether violence was experienced as a victim or as a witness. In fact, the relationship between community violence and psychopathology was found to be generally nonsignificant regardless of social support status. CONCLUSIONS: These findings suggest that exposure to family violence may affect development differently than exposure to community violence, allowing social support to effectively buffer the effects of family, but not community violence. This finding highlights the importance of examining violence exposure that occurs within the family separately from violence exposure that occurs within the community. PMID- 10798838 TI - Evaluations of child transgressions, disciplinary choices, and expected child compliance in a no-cry and a crying infant condition in physically abusive and comparison mothers. AB - OBJECTIVE: Several components of a social information processing model of child physical abuse were tested. Abusive and comparison mothers' evaluations of children's transgressions, choices of disciplinary techniques, expectations for children's compliance following discipline, and appraisals of the appropriateness of disciplinary choice were examined in a no-cry and a crying-infant condition. METHOD: Thirty physically abusive and 30 matched comparison mothers were individually matched on ethnic background, age, education, marital status, number of children, and cognitive ability. Mothers were asked to respond to questions related to vignettes describing children engaging in moral, conventional, and personal transgressions. RESULTS: As predicted, abusive, relative to comparison, mothers evaluated conventional and personal, but not moral, transgressions as more wrong, used more power assertion (physical and verbal force), expected less compliance from their own children, and appraised their own disciplinary responses as less appropriate. In contrast to expectations, there were no group by cry condition interaction effects on any of the study measures. CONCLUSIONS: The findings provide additional support for the view that abusive, relative to comparison, mothers are different in their evaluations and expectations of their own children's behaviors and that they more frequently select aversive disciplinary techniques. However, given the lack of an expected differential impact of a stressful condition on the cognitions and disciplinary choices in abusive mothers, additional research is needed. PMID- 10798837 TI - Children in foster care: factors influencing outpatient mental health service use. AB - OBJECTIVE: To determine factors influencing outpatient mental health service use by children in foster care. METHOD: Detailed survey and administrative data were collected on 480 children who entered long-term foster care in San Diego County from May 1990 through October 1991. These data were linked with claims data from Medicaid and San Diego County Mental Health Services information systems. A Poisson regression model was used to determine whether the following factors influenced outpatient mental health service use: age, race/ethnicity, gender, maltreatment history, placement pattern, and behavioral problems as measured by the Achenbach Child Behavior Checklist (CBCL). RESULTS: Except for maltreatment history, all independent variables included in the multivariate regression model were statistically significant. The total number of outpatient mental health visits increased with age, male gender, and non-relative foster placements. Relative to Caucasians, visits were lower for Latinos, and Asian/Others, but comparable for African-Americans. Concerning maltreatment history, differences were only found in one category; children experiencing caretaker absence received fewer visits compared to children who did not experience caretaker absence. Children with CBCL Total Problem Scale T-scores of 60 or greater had significantly more visits than those with a score less than 60. CONCLUSIONS: Both clinical and non-clinical factors influence outpatient mental health service use by foster children. Limitations imposed by gender, race/ethnicity, and placement setting need to be addressed by child welfare policies. These finding suggest that guidelines are needed to systematically link children in foster care with behavioral problems to appropriate services. PMID- 10798839 TI - Impact of child noncompliance on stress appraisals, attributions, and disciplinary choices in mothers at high and low risk for child physical abuse. AB - OBJECTIVE: The study investigated the impact of repeated child noncompliance on stress appraisals, attributions, and disciplinary choices in high- and low-risk mothers. METHOD: Fifty (25 high-risk and 25 demographically, matched low-risk) mothers responded to questions related to stress appraisals, attributions, and disciplinary choices following presentations of a child engaging in repeated noncompliance. RESULTS: After repeated child noncompliance, high-risk, compared to low-risk, mothers perceived more threat and uncontrollability, rated child behaviors as more stressful, and reported higher levels of negative affect. High risk mothers also reported more stable, global, and intentional attributions, with a trend toward more internal attributions, but did not differ in their evaluations of wrongness and seriousness of the child's behavior. After repeated noncompliance, a risk group difference was found in estimates of future child compliance but not in the use of power assertive discipline. CONCLUSIONS: Results support the view that high-risk, relative to low-risk, mothers are differentially responsive to stressful situations and differ in their attributions for negative child behaviors and in their expectations of future child compliance. However, since risk group differences in disciplinary choices were not also found, additional research is needed to demonstrate the process through which risk group cognitive and affective differences are related to differences in disciplinary behavior. PMID- 10798840 TI - From maltreatment report to juvenile incarceration: the role of child welfare services. AB - OBJECTIVE: This study examined whether children who received child welfare services (e.g., in-home or out-of-home placement) were more or less likely to become incarcerated as serious and violent youthful offenders than those children who were investigated as victims of abuse and neglect but received no further child welfare intervention. METHOD: Administrative data on child abuse reporting, foster care, birth records, and juvenile corrections (CYA) were linked to prospectively examine the risk of incarceration as an adolescent following an investigation of abuse or neglect after age 6. The 10 county California sample included 159,549 school-aged children reported for abuse and neglect after 1990. RESULTS: About 8 per 1,000 children in the sample were later incarcerated in CYA. African American and Hispanic children who received in-home or foster care services after the index investigation event had a lower risk of incarceration than those whose cases were closed after the investigation. Among females, the rate of incarceration was highest for those who experienced foster or group care placements. Children initially reported for neglect were more likely to be incarcerated than those reported for physical or sexual abuse. CONCLUSIONS: Public child welfare services have rarely been assessed in terms of future negative child outcomes. This study finds that one serious negative outcome, CYA involvement, can only be understood when a number of factors are considered. The importance of understanding the differences between how different subpopulations respond to services is highlighted. Specifically, our findings suggest that more attention should be focused on children who are now receiving no services after an investigated child abuse and neglect report, on females, and on victims of child neglect. PMID- 10798841 TI - A profile of the oro-facial injuries in child physical abuse at a children's hospital. AB - OBJECTIVE: The aim of the present study was to determine the head, face and neck injuries associated with child abuse cases in the Cape Peninsula, Cape Town. South Africa. METHOD: A retrospective, record-based analysis (n = 300) of non accidental injuries at a Children's Hospital over a 5-year period (1992-1996) was carried out. RESULTS: The mean age of the sample was 4.75 years--54.3% were boys and 45.7% were girls. Most of the crimes were committed in the child's own home (88.7%). Crimes were reported by mothers (48.7%), grandmothers (11.7%) and day hospitals (13%). Ninety percent of the perpetrators were known to the victim. The majority of the perpetrators were male (79%)--20% the perpetrators were the mother's boyfriend; 36% the father or step father, and in 12% the mother was responsible. Thirty-five percent of perpetrators were under the influence of alcohol or drugs when they committed the offence: 64.7% of cases suffered serious injuries, 48.7% had to be hospitalized, four children were critically injured and died. The head, face, neck, and mouth were the sites of physical injury in 67% of the 300 cases reviewed. The face was the most frequently injured (41%) part of the body, with the cheek being the most common site for the injury. The range and diversity of the oro-facial injuries included skull fractures, subdural hematomas, retinal hemorrhages, bruises, burns, and lacerations. Injuries to the mouth included fractured teeth, avulsed teeth, lacerations to the lips, frenum, tongue, and jaw fractures. CONCLUSIONS: The main conclusions of this study were (i) under 2-year-old children were most at risk from abuse (36%); (ii) the number of the reported injuries to the oral cavity was extremely low (11%); and (iii) no dentists participated in the examination of any of the patients. Intra-oral injuries may be overlooked because of the medical examiner's unfamiliarity with the oral cavity. Oral health professionals should be consulted for diagnosis, advice and treatment. PMID- 10798842 TI - A study of clerics who commit sexual offenses: are they different from other sex offenders? AB - OBJECTIVE: The goal of this study was to determine if cleric-sex offenders differed significantly from other sex offenders when compared to a control group and assessed with standard instruments that examine the major factors important in sexual offenses. METHOD: Twenty-four male clerics accused of sexual offenses were compared to 24 male sex offender controls, matched on offense type, age, education, and marital status. Both groups were compared to a general sample of sex offenders (n = 2125) matched only for offense type. The three groups were compared on sexual history and preference, substance abuse, mental illness and personality, history of crime and violence, neuropsychological impairment, and endocrine abnormalities, using reliable and valid measures. RESULTS: The clerics in this study formed a statistically significant highly educated, older, and predominantly single subgroup of sex offenders. The majority of cleric-sex offenders suffered from a sexual disorder (70.8%), predominantly homosexual pedophilia, as measured by phallometric testing, but did not differ from the control groups in this respect. The clerics were comparable to the other two groups in most respects, but tended to show less antisocial personality disorders and somewhat more endocrine disorders. The most noteworthy features differentiating the clerics from highly educated matched controls were that clerics had a longer delay before criminal charges were laid, or lacked criminal charges altogether, and they tended to use force more often in their offenses. CONCLUSIONS: In spite of differences in age, education, and occupation between cleric-sex offenders and sex offenders in general, the same procedures should be used in the assessment of this group as for the sex offender population in general. Hypotheses about reduced sexual outlet and increased sexual abuse of clerics in childhood were not supported. Assessment and treatment of cleric-sex offenders should focus especially on sexual deviance, substance abuse, and endocrine disorders. PMID- 10798843 TI - Chronic pain and health care utilization in women with a history of childhood sexual abuse. AB - OBJECTIVE: This cross-sectional controlled study investigated the association between chronic pain, health care utilization and a history of childhood sexual abuse. SUBJECTS: Three groups, constituting 80 women in total, were studied (1) attendees at group therapy for individuals who had experienced childhood sexual abuse (n = 26); (2) Two control groups consisting of nonabused (a) psychiatric outpatients (n = 33); and (b) nurses (n = 21). SETTING: The setting was a university affiliated community and tertiary care hospital in London. Ontario. OUTCOME MEASURES: Each subject voluntarily completed questionnaires documenting history of childhood abuse, pain, psychological symptomatology and medical and surgical history. RESULTS: Sixty-nine percent of the women who had experienced childhood sexual abuse reported a chronic painful condition lasting more than three months, compared to 43% of the combined control groups (p = .026). Women who had experienced childhood sexual abuse reported a greater number of painful body areas (p = .003), more diffuse pain and more diagnoses of fibromyalgia (p = .013). They had more surgeries (p = .037), hospitalizations (p = .0004) and family physician visits (p = .046). CONCLUSIONS: Women with a history of childhood sexual abuse reported more chronic pain symptoms and utilized more health care resources compared to nonabused control subjects. Identification of such a history in the patient experiencing persisting pain may be the first step toward a successful combination of medical and psychosocial interventions. PMID- 10798844 TI - A comparison of sexually abused and non-sexually abused adolescents in a clinical treatment facility using the MMPI-A. AB - OBJECTIVE: In the current study, the adolescent version of the Minnesota Multiphasic Personality Inventory, the MMPI-A, was used to address concerns voiced about the mixing of different developmental ages, use of inadequate comparison groups, and the lack of reliance on reliable and valid measures of psychological, behavioral, and psychosocial problems used in previous studies of sexually abused children and adolescents. METHOD: A total of 107 adolescents in residential treatment (73 boys and 34 girls) were included in the study. Seventy two of these adolescents had reportedly been sexually abused (42 boys and 30 girls); 35 had not been abused (31 boys and 4 girls). Predictions based on previous research with adolescents were made and tested regarding which MMPI-A validity, clinical, and content scales would differ between the sexually abused and non sexually abused groups of adolescents. RESULTS: Overall, and consistent with many predictions, sexually abused adolescents had both statistically and clinically higher elevations on several MMPI-A scales than did their non-abused counterparts. No scales were more elevated for non-abused adolescents than for abused adolescents. CONCLUSIONS: Sexually abused adolescents in residential treatment, as a group, present with concerns that their non-abused counterparts did not have, or did not share to the same extent. Treatment recommendations based on the MMPI-A scale elevations are provided, limitations of the current study discussed, and directions for further research are suggested. PMID- 10798845 TI - A matched cohort comparison of a criminal justice system's response to child sexual abuse: a profile of perpetrators. AB - OBJECTIVE: This study was undertaken to examine whether alleged child sexual abuse perpetrators are handled differently by the courts than other alleged felony perpetrators. Comparisons were made from the time of prosecutorial case acceptance through prosecution to sentencing, with emphasis on judicial and prosecutorial decision-making and plea-bargaining. METHOD: Data were retrospectively abstracted on the entire defendant population of cases of sexual abuse of children and adolescents (ages 2-17) over a 5-year period. Using a case flow analysis, comparisons were made between a child sexual abuse cohort and a cohort of matched felony cases from a single jurisdiction. RESULTS: Three important findings emerged. First, compared to other felons, abuse perpetrators were employed, had been married, were mostly European American, and were older than 30 years of age. Second, in the abuse cohort, as many as 14% had a previous sexual or violent record compared to 2% in the comparison group. Third, similar percentages of perpetrators in both groups were released on their own recognizance, had the charges against them dropped, and were found guilty. As well, no differences between groups were found in the proportion of individuals sentenced to jail, probation, counseling, or work release. CONCLUSIONS: Although the treatment of perpetrators of child sexual abuse was similar to the treatment of perpetrators of other felonies, the profile of the child abuse perpetrator was quite different. Knowledge about this profile may impact prosecution or treatment and recidivism rates, to the extent that recidivism is related to characteristics of the abuse perpetrator. PMID- 10798846 TI - Multi-criminal and violent groups among child sex offenders: a heuristic typology in a 2-year cohort of 374 men in two English counties. AB - OBJECTIVE: To determine the lifetime criminality of men charged with a sexual offence against a child and to identify any heuristic typologies. METHOD: The police files of a 2-year cohort of 462 men charged with a sexual offence against a minor (<17) in two English counties were analyzed; 374 were convicted. The patterns of their convictions produced a typology based upon judicially substantiated parameters. RESULTS: Half the guilty men had prior convictions for non-sexual crimes, including 24% with convictions for violent offences. Three heuristic groups emerged: "Sex-Only" offenders (S.O., n = 186); "Sex And Other" offenders (S.A.O., n = 97); and "Sex and Violent" offenders (S.V., n = 91), averaging three sex convictions each. However, the S.A.O. and S.V. men had more non-sexual than sex convictions, averaging 5.1 versus 10.5, justifying the designation of "multi-criminal" child sex abusers. However, the patterns of these men's sexual offending did not differ, except that the S.O. men's victims were significantly more often intra-family, 36%, 31%, and 13%, respectively; indicating that the majority of these men's victims were extra-familial (71%). Significantly, 19% of the convicted men had at least one "not guilty" verdict for a sex charge, as opposed to 4% for non-sexual crimes. CONCLUSIONS: While the S.O. group appear to consist of traditional types of fixated and regressed offenders, in men charged with offences, they are atypical of most sex abusers since their victims are mainly extra-familial. These "multi-criminal" men present particular challenges for custodial and therapeutic settings, and the potential dangerousness of these offenders should not be underestimated. PMID- 10798847 TI - Proteolytic signals from Magdeburg. PMID- 10798848 TI - Mindful immunology: neuroimmunomodulation. PMID- 10798849 TI - Preparing the ground for vaccination against multiple myeloma. PMID- 10798850 TI - Systemic adjuvant treatment for premenopausal node-negative breast cancer. The International Breast Cancer Study Group. PMID- 10798851 TI - [New forms of hepatobiliary complications in chronic intestinal inflammatory disease]. PMID- 10798852 TI - Teen pregnancy reduction in program with peer staff. PMID- 10798853 TI - Case-based teaching in adolescent medicine. PMID- 10798854 TI - Congratulations on the 20th anniversary of the Journal of Adolescent Health!!! PMID- 10798856 TI - Proceedings of the VIIth International Symposium on the Facial Nerve. Cologne, Germany, June 9-14, 1992. PMID- 10798855 TI - A latent trait finite mixture model for the analysis of rating agreement. AB - This article presents a latent distribution model for the analysis of agreement on dichotomous or ordered category ratings. The model includes parameters that characterize bias, category definitions, and measurement error for each rater or test. Parameter estimates can be used to evaluate rater performance and to improve classification or measurement with use of multiple ratings. A simple maximum likelihood estimation procedure is described. Two examples illustrate the approach. Although considered in the context of analyzing rater agreement, the model provides a general approach for mixture analysis using two or more ordered caregory measures. PMID- 10798858 TI - The Transgenic Animals Research Conference. Tahoe City, California, USA. 15-19 August 1999. Abstracts. PMID- 10798857 TI - 7th International HUGO Mutation Database Meeting, October 19, 1999, San Francisco, USA. AB - The 7th International HUGO Mutation Database Meeting was held on October 19, 1999 in conjunction with the annual meeting of the American Society of Genetics in San Fransisco, California, U.S.A. Meeting highlights are described, including discussions of topics such as the ethical aspects of variation databases, ethical guidelines which should be established immediately, data protection laws which may affect access to data, and plans to make variation databases financially self sustaining. A resolution was passed which encourages HUGO and Mutation database Initiative (MDI) collaboration (under the name HUGO-MDI) to provide an integrated, properly funded system of variation databases. PMID- 10798859 TI - Working with Paul Sigler. PMID- 10798860 TI - Paul Sigler: a structural biologist with passion. PMID- 10798861 TI - Towards a synthetic pneumococcal vaccine: synthetic oligosaccharides as tools for improving the specificity of enzyme-linked immunosorbent assays. PMID- 10798862 TI - Comparison of mutant and wild-type p53 proteins in Merkel cell carcinoma. PMID- 10798863 TI - Malaria in the French West Indies. PMID- 10798864 TI - Smoking among Buddhist monks in Phnom Penh, Cambodia. PMID- 10798865 TI - Effect of smokefree bar law on bar revenues in California. PMID- 10798866 TI - Passive smoking and an increased risk of acute stroke. PMID- 10798867 TI - A tentative illustration of the smoking initiation and cessation cycles. PMID- 10798868 TI - Identification of ider[17q] in addition to t[15;17] in acute promyelocytic leukemia using whole chromosome painting probes made by interspecies hybrid using inter-Alu PCR. PMID- 10798869 TI - 2nd International Conference on HIV and Iron. Brugge, Belgium, March 31-April 1, 2000. Abstracts. PMID- 10798870 TI - Reappraisal of old operation methods. PMID- 10798871 TI - Environmental medicine, part 2 - health effects of and protection from ubiquitous airborne solvent exposure. AB - Chemicals known as solvents are part of a broad class of chemicals called volatile organic compounds. These compounds are used in a variety of settings, are ubiquitous, and off-gas readily into the atmosphere. Asa result of their overuse, they can be found in detectable level virtually all samples of both indoor and outdoor air. Certain of these compounds are detectable in adipose samples of all U.S. residents Once in the body they can lead to a variety of neurological, immunological, endocrinological, genitourinary, and hematopoietic problems. Some individuals also have metabolic defects that diminish the liver's clearing capacity for these compounds. Supplementation may be of benefit to help clear these compounds from the body and to prevent adverse health effects. PMID- 10798872 TI - Literature review. PMID- 10798873 TI - Gene therapy and tissue engineering patents abound. PMID- 10798875 TI - Web alert. Cell regulation. PMID- 10798874 TI - Paper alert. Microbiology. PMID- 10798876 TI - Prevalence of benign epileptiform variants and its implications in the management of epilepsy. PMID- 10798877 TI - Stages of adjustment to a medical diagnosis of a serious somatic condition. AB - Diagnosis of a serious somatic condition may precipitate a seven stage coping process: shock, denial, procrastination, bargaining, depression, anger and adaptation. Psychometric/ statistical methods for operationalizing the stages are suggested. 1999 Elsevier, Paris. PMID- 10798879 TI - Prison suicides in Italy in 1996-1997. PMID- 10798878 TI - Liaison-psychiatry: reasons for consultation, diagnosis, and differences between medical and surgical patients. PMID- 10798880 TI - 'Infectious web'. PMID- 10798881 TI - Hydration of fat-fre body mass: new physiological modeling approach. PMID- 10798882 TI - Interpretation of muscle damage from fixed tissue obtained by needle biopsy. PMID- 10798883 TI - Intracellular Po2 and bioenergetic measurements in skeletal muscle: the role of exercise paradigm. PMID- 10798884 TI - HHS award $527 million for AIDS CARE. PMID- 10798885 TI - HIV receptor gene is patented. PMID- 10798886 TI - Sustiva granted full FDA approval. PMID- 10798887 TI - National urine-based HIV and STD testing service. PMID- 10798888 TI - Glaxo supports in-prison programs. PMID- 10798889 TI - HIV drug resistance testing. PMID- 10798890 TI - HIV reference laboratory opens in Botswana. PMID- 10798891 TI - [Comment on M. Dettenkofer et al.: Disposable versus reuse of surgical drapes in the operating room]. PMID- 10798892 TI - Follow up of a HNF-1alpha mutant carrier with a severely impaired beta cell function. PMID- 10798893 TI - Mild traumatic brain injury and the postconcussive syndrome. PMID- 10798894 TI - Accidental injection of thiopental into the epidural space. PMID- 10798895 TI - 'Diprivan': a change of formulation. PMID- 10798896 TI - Electromyographic quantification of the paralysing effect of botulinum toxin in the sternocleidomastoid muscle. AB - The effect of botulinum toxin (BT) upon the human body has so far been measured by using clinical scales monitoring its overall therapeutic effect upon the disorders treated. Clinical scales, however, usually lack sensitivity, are rarely validated and are integrating a number of uncontrollable parameters. After validation of the methodology in a group of 10 controls, we investigated the BT induced amplitude reduction of the maximal voluntary electromyographic activity (M-EMG amplitude reduction) of the sternocleidomastoid muscle in a group of 34 patients with cervical dystonia undergoing regular BT therapy with Botox (Allergan, Irvine, Calif., USA; n = 16) or Dysport (Ipsen, Maidenhead, UK; n = 18). With Botox doses of 20 mouse units the M-EMG amplitude reduction was 80% (SD = 3.9%, n = 4), with 40 it was 84% (SD = 10.8%, n = 4), with 60 it was 85% (SD = 2.6%, n = 2) and with 80 it was 91% (SD = 5.8%, n = 6). With Dysport doses of 100 mouse units the M-EMG amplitude reduction was 70% (SD = 7.6%, n = 4), with 200 it was 85% (SD = 10.4%, n = 5), with 300 it was 83% (SD = 9.2%, n = 3), with 400 it was 78% (SD = 6.7%, n = 3) and with 500 it was 91% (SD = 5.8%, n = 5). The methodology presented can measure M-EMG amplitude reductions with a precision of about 10%. Dose-efficacy relationships can be used for dose optimisation, evaluation of BT therapy failure and comparison of different preparations and types of BT. PMID- 10798897 TI - Literature and the arts in medical education. PMID- 10798898 TI - Spirituality in residency education: taking the next step. PMID- 10798899 TI - Goal setting for community preceptorships. PMID- 10798900 TI - Along for the ride. PMID- 10798901 TI - Towards immunotherapy for pancreatic cancer. PMID- 10798902 TI - Ring-like elevations in the large bowel: endoscopic signs to distinguish the artefact from true neoplastic lesions. PMID- 10798903 TI - Liver biopsy under ultrasound control: implications for training. PMID- 10798904 TI - Recent publications in hematological oncology. PMID- 10798905 TI - Novel immunological aspects of CMV-related disease: pathogenesis, diagnosis, and therapy. Proceedings of the 2nd Symposium on CMV-Related Immunopathology. Maastricht, The Netherlands. September 17-18, 1999. PMID- 10798906 TI - The Agency for Healthcare Research and Quality: new challenges, new opportunities. PMID- 10798907 TI - Slowly remaking the U.S. Healthcare system. PMID- 10798908 TI - Death of Dr. Still. PMID- 10798909 TI - Regarding Cieki et al. IJROBP 1999;45(4):963-968. PMID- 10798910 TI - Pathologic examination of resected specimen. PMID- 10798911 TI - The Rorschach test in clinical diagnosis--a brief commentary. PMID- 10798912 TI - Limitations of the Rorschach as a diagnostic tool: A reply to Garfield (2000), Lerner (2000), and Weiner (2000) PMID- 10798913 TI - A modified JAM assay detects apoptosis induced in activated lymphocytes by FasL+ human adherent tumor cells. PMID- 10798914 TI - Antiprogestins RU486 and ZK299 suppress basal and LHRH-stimulated FSH and LH secretion at pituitary level in the rat in an oestrous cycle stage-dependent manner. AB - We have previously shown that administration of antiprogestin (AP) type II RU486 to ovariectomized (OVX) rats on the morning of pro-oestrus decreases the magnitude of preovulatory gonadotrophin surge. This suggests that the effect of RU486 on LHRH-dependent gonadotrophin release may be independent of its ability to block progesterone actions. The aim of the present research was to study the possible site of RU486 action and to determine whether the gonadotrophin suppressive effect of APs RU486 and ZK299 is dependent on the oestrogen background. Intact or OVX rats in the morning of pro-oestrus were injected s.c. with 4 mg of RU486 or ZK299 (AP type I) at 0900 h on pro-oestrus. At 1830 h, serum concentration of FSH and LH and median eminence (ME) content of LHRH were determined. In the second experiment, the effect of RU486 and ZK299 on pituitary responsiveness to LHRH (100 ng, i.p.) and ME content of LHRH at 1830 h pentobarbital-blocked intact or OVX rats was evaluated. In the last study, the anterior pituitary release of FSH and LH from pro-oestrus or metoestrus donors incubated with or without LHRH (1, 10 or 100 nM) in the presence or absence of APs (20 nM) was evaluated. Both APs reduced serum FSH and LH levels at 1830 h on pro-oestrus in intact and OVX rats. The suppressive effect on gonadotrophin release brought about by AP treatment was also evidenced in PB-blocked intact and OVX rats. This suggested that the inhibitory effect of APs occurred, at least in part, at pituitary level. Furthermore, in the absence of the natural ligand, APs significantly reduced basal and LHRH-stimulated FSH and LH release from pro oestrous but not from metoestrus pituitaries. In conclusion, these experiments have shown, both 'in vivo' and 'in vitro', that APs RU486 and ZK299 have suppressive effects at pituitary level on basal and LHRH-stimulated FSH and LH secretion, regardless of their antiprogestagenic activity, in pro-oestrus but not in metoestrus. PMID- 10798915 TI - Diagnostic re-evaluation of a case of 'cerebellar atrophy with Huntington's disease'. PMID- 10798916 TI - Larger debate underlies spiral CT screening for lung cancer. PMID- 10798917 TI - Antibiotics for bacterial prostatitis. PMID- 10798918 TI - Female orthotopic bladder substitution: a good operation in the right circumstances. AB - PURPOSE: We present the long-term results of orthotopic bladder substitution in women. MATERIALS AND METHODS: We prospectively assessed 15 female patients undergoing orthotopic bladder substitution between October 1984 and April 1998. The indication for diversion was bladder carcinoma in 8 cases, gynecologic malignancy in 2 and benign disease in 5. RESULTS: At a median followup of 19 months daytime and nighttime continence was 100% and 80%, respectively. All but 1 patient voided spontaneously. Two patients who were initially able to void successfully had voiding difficulties in the initial 10 months due to prominent mucosal tissue at the bladder neck. In each case this condition was successfully managed by endoscopic resection. There has been no local recurrence of bladder carcinoma. CONCLUSIONS: With strict selection criteria and vigilant followup female orthotopic bladder substitution produces excellent long-term functional results comparable to those in men. However, the limits in women who require radical cystectomy for invasive bladder cancer cannot be determined from our results. Further long-term data on the local recurrence rate of invasive urothelial malignancy are needed before the true risk may be quantified accurately. PMID- 10798919 TI - Hypoxic radiosensitizers in radical radiotherapy for patients with bladder carcinoma: hyperbaric oxygen, misonidazole, and accelerated radiotherapy, carbogen and nicotinamide. PMID- 10798920 TI - A phase II trial of methotrexate, cisplatin, 5-fluorouracil, and leucovorin in the treatment of invasive and metastatic urothelial carcinoma. PMID- 10798922 TI - A new generation of semirigid fiberoptic ureteroscopes. PMID- 10798923 TI - Nephroscopy sheath characteristics and intrarenal pressure: human kidney model. PMID- 10798921 TI - Neoadjuvant cisplatin, methotrexate, and vinblastine chemotherapy for muscle invasive bladder cancer: a randomised controlled trial. PMID- 10798924 TI - In vitro results with special plastics for biodegradable endoureteral stents. PMID- 10798925 TI - [Johann Mikulitch (150-th anniversary)]. PMID- 10798926 TI - Innovations in medical education. PMID- 10798927 TI - GP registrars need training in child abuse and neglect. PMID- 10798928 TI - Teacher development in hospitals--where should we go? PMID- 10798929 TI - 'I wished I was the patient'. An evaluation of a complementary medicine module for third year medical students. PMID- 10798930 TI - [New classes of drug for treating HIV infections]. PMID- 10798931 TI - The ABCs of cystic fibrosis. PMID- 10798932 TI - Death and antimutagenicity. PMID- 10798933 TI - Medicine in the past millennium. PMID- 10798934 TI - Medicine in the past millennium. PMID- 10798935 TI - Medicine in the past millennium. PMID- 10798936 TI - Medicine in the past millennium. PMID- 10798937 TI - Medicine in the past millennium. PMID- 10798938 TI - Medicine in the past millennium. PMID- 10798939 TI - Medicine in the past millennium. PMID- 10798940 TI - Medicine in the past millennium. PMID- 10798941 TI - Reducing neonatal group B streptococcal disease. PMID- 10798942 TI - Insertion of femoral-vein catheters for practice during cardiopulmonary resuscitation. PMID- 10798943 TI - Insertion of femoral-vein catheters for practice during cardiopulmonary resuscitation. PMID- 10798944 TI - Insertion of femoral-vein catheters for practice during cardiopulmonary resuscitation. PMID- 10798945 TI - Insertion of femoral-vein catheters for practice during cardiopulmonary resuscitation. PMID- 10798946 TI - Executives with white coats--managed-care medical directors. PMID- 10798947 TI - Executives with white coats--managed-care medical directors. PMID- 10798948 TI - Executives with white coats--managed-care medical directors. PMID- 10798949 TI - Executives with white coats--managed-care medical directors. PMID- 10798950 TI - Anaphylaxis from inulin in vegetables and processed food. PMID- 10798951 TI - 10th Italian Congress of Neuroepidemiology. Taormina, March 23-26, 2000. Abstracts. PMID- 10798952 TI - Cerebellar and medullar histoplasmosis. PMID- 10798953 TI - Better late than never. PMID- 10798954 TI - [Polina Andreevna Petrishcheva and the importance of her works in the development of parasitology (on the centenary of her birth)]. PMID- 10798955 TI - [The 60th anniversary of the creation of the theory of the natural foci of diseases]. PMID- 10798956 TI - [An evaluation of male reproductive success by body size in swarms of Culex pipiens pipiens (Diptera: Culicidae)]. PMID- 10798957 TI - [A new species of myobiid mite Radfordia cricetuliphila sp. n. (Acari: Myobiidae) from the striped hamster Cricetulus barabensis (Rodentia: Cricetidae) from Buryatia]. PMID- 10798958 TI - Sodium channels in the cytoplasm of Schwann cells. PMID- 10798959 TI - Summer 1999: caring, death, and good doctors. PMID- 10798960 TI - Summer 1999: caring, death, and good doctors. PMID- 10798962 TI - Milestones for training safe doctors who are good doctors. PMID- 10798961 TI - Summer 1999: caring, death, and good doctors. PMID- 10798963 TI - The radioresponse of the central nervous system: a dynamic process. AB - Radiation continues to be a major treatment modality for tumors located within and close to the central nervous system (CNS). Consequently, alleviating or protecting against radiation-induced CNS injury would be of benefit in cancer treatment. However, the rational development of such interventional strategies will depend on a more complete understand-ing of the mechanisms responsible for the development of this form of normal tissue injury. Whereas the vasculature and the oligodendrocyte lineage have traditionally been considered the primary radiation targets in the CNS, in this review we suggest that other phenotypes as well as critical cellular interactions may also be involved in determining the radio-response of the CNS. Furthermore, based on the assumption that the CNS has a limited repertoire of responses to injury, the reaction of the CNS to other types of insults is used as a framework for modeling the pathogenesis of radiation-induced damage. Evidence is then provided suggesting that, in addition to acute cell death, radiation induces an intrinsic recovery/repair response in the form of specific cytokines and may PMID- 10798964 TI - Toxicity of ALS-linked SOD1 mutants. PMID- 10798965 TI - Patent prompts Rochester to sue for slice of drug profits. PMID- 10798966 TI - Paleoanthropology. Is Alexander the Great's father missing, too? PMID- 10798967 TI - German vote. Animal rights amendment defeated. PMID- 10798968 TI - Evolution. When fittest survive, do other animals matter? PMID- 10798969 TI - Collaborative research. Plans for Mars unite cancer, space agencies. PMID- 10798971 TI - Paleontology. Revealing a dinosaur's heart of stone. PMID- 10798970 TI - Psychiatry. Are placebo-controlled drug trials ethical? PMID- 10798972 TI - Human genome. DOE team sequences three chromosomes. PMID- 10798973 TI - Science education. Ehlers offers remedies for ailing U.S. system. PMID- 10798975 TI - American Chemical Society meeting. Chemists unveil molecular wizardry in San Francisco. PMID- 10798974 TI - Society of Toxicology meeting. Hazards of particles, PCBs focus of Philadelphia meeting. PMID- 10798976 TI - Plant breeding. Hopes grow for hybrid rice to feed developing world. PMID- 10798977 TI - Physical hazards of tailings dams. PMID- 10798978 TI - Historical views on the human embryo. PMID- 10798980 TI - Shape persistence of synthetic polymers. PMID- 10798979 TI - HIV-associated dementia. PMID- 10798981 TI - Movement patterns in spoken language. PMID- 10798982 TI - Circadian rhythms. Marking time for a kingdom. PMID- 10798983 TI - Nota bene. Contortions of the heart. PMID- 10798985 TI - NIH nomination on hold for this year. PMID- 10798984 TI - In contrast to Dolly, cloning resets telomere clock in cattle. PMID- 10798986 TI - National academies. Task force tinkers with research council. PMID- 10798987 TI - Global warming. Draft report affirms human influence. PMID- 10798989 TI - Endangered species. CITES puts off plan to hasten shipments. PMID- 10798988 TI - South Africa. AIDS researchers decry Mbeki's views on HIV. PMID- 10798990 TI - Biomedical policy. NIH, under pressure, boosts minority health research. PMID- 10798991 TI - Working in the hot zone: Galveston's microbe hunters. PMID- 10798992 TI - National Science Foundation. Information technology takes a different tack. PMID- 10798993 TI - Nuclear Waste disposal. Science and policy clash at Yucca Mountain. PMID- 10798994 TI - Astrobiology science conference. The science of astrobiology takes shape. PMID- 10798995 TI - Opportunity for agricultural biotechnology. PMID- 10798996 TI - Support for Congolese conservationists. PMID- 10798997 TI - BIOPAT does not trade in names. PMID- 10798998 TI - Metabolic analysis in drug discovery. PMID- 10798999 TI - Sulfate clues for the early history of atmospheric oxygen. PMID- 10799000 TI - Gene therapy. The best of times, the worst of times. PMID- 10799001 TI - Neurobiology. Of flies and mice. PMID- 10799003 TI - Proceedings of the XXXVII European Congress of Toxicology. EUROTOX '99. Oslo, Norway, June 27-30, 1999. PMID- 10799002 TI - Structural biology. Light at the end of the channel. PMID- 10799005 TI - Is there a cure for aplastic anaemia? In memoriam Bruno Speck 1934-1998. PMID- 10799004 TI - 49,XXXXY syndrome with hydronephrosis caused by intravesical ureterocele. AB - A 1-month-old boy was referred to our hospital with right hydronephrosis. Excretory urography showed poor visualization of the right kidney and a filling defect in the bladder. Chromosomal analysis of peripheral blood revealed a karyotype of 49,XXXXY, and a diagnosis of 49,XXXXY Klinefelter s syndrome associated with hydronephrosis caused by intravesical ureterocele was made. 49,XXXXY Klinefelter s syndrome with anomalies of the urinary tract is extremely rare, and only 2 cases have been reported so far. PMID- 10799006 TI - Living with severe aplastic anaemia for 22 years. A patient tells his story (supplemented with information from the Hematologic Clinic Basel, Switzerland). PMID- 10799007 TI - Bruno Speck in Leiden: the clinician, the scientist, the man. PMID- 10799008 TI - An invasive isolate of Neisseria meningitidis showing decreased susceptibility to quinolones. PMID- 10799010 TI - Annual Scientific Meeting of the Urological Society of Australasia. Adelaide, March 1999. Abstracts. PMID- 10799009 TI - Antiretroviral-drug concentrations in semen. PMID- 10799011 TI - Sir William Manchester Memorial Lecture 1999: the influence of one man on plastic and reconstructive surgery in New Zealand. PMID- 10799012 TI - Should reductase inhibitor therapy to lower cholesterol be instituted in the setting of an acute coronary event? PMID- 10799013 TI - The in vivo oxidized LDL to LDL-cholesterol ratio is correlated with lipid content of LDL. PMID- 10799014 TI - A common lecithin: cholesterol acyltransferase gene variant (Ser208-->Thr) PMID- 10799015 TI - Serum homocysteine, folate and thermolabile variant of MTHFR in healthy Sri Lankans living in London. PMID- 10799016 TI - Glycemia, MTHFR genotype and low homocysteine in uncomplicated type 2 diabetic patients. PMID- 10799017 TI - Does vitamin E beneficially affect muscle pains during HMG-Co-enzyme-A-reductase inhibitors without CK-elevation? PMID- 10799018 TI - Circadian variation of endothelium-dependent vasodilatation of the brachial artery as a confounding factor in the evaluation of endothelial function. PMID- 10799049 TI - Live oral poliovirus vaccines do not contain detectable simian virus 40 (SV40) DNA. AB - Prior to 1962, poliovirus vaccines produced in rhesus monkey kidney cells were contaminated with SV40. Recent studies reporting the detection of SV40 in human tumours raised concern that SV40 may be oncogenic in humans. To provide further assurance that currently used poliovirus vaccines are not contaminated with SV40, we used the polymerase chain reaction (PCR) to search for SV40 DNA in live oral poliovirus vaccines manufactured in the United States between 1972 and 1996. SV40 DNA sequences were not found in any of the vaccine lots tested. PMID- 10799050 TI - Viral inactivation of intramuscular immune serum globulins. AB - A new process for the production of intramuscular immunoglobulin products is described which includes viral inactivation through solvent-detergent treatment. Removal of solvent-detergent was accomplished by precipitation, filtration and diafiltration. Process-scale preparations had appropriate antibody potency levels, and improved IgG integrity relative to traditional IGIM products. Moreover, acceptable results were obtained in all in vitro and in vivo pre clinical toxicology testing, as well as clinical evaluation. Scaled-down experiments demonstrated that the new process provides effective viral inactivation. Taken together, these results indicate that the new products should have the same efficacy of the previous IGIM products albeit with safety and processing improvements. PMID- 10799051 TI - Use and validation of an NMR test for the identity and O-acetyl content of the Salmonella typhi Vi capsular polysaccharide vaccine. PMID- 10799052 TI - The maintenance of challenge strains used in the potency test for canine leptospira vaccines. AB - The challenge test for leptospira vaccines required by most licensing authorities is difficult to standardise and unreliable. One of the main contributory factors to this, is the difficulty in maintaining the virulence of the challenge strain. This paper describes work carried out to assess the practicality of storing challenge strains of Leptospira canicola and Leptospira icterohaemorrhagiae in liquid nitrogen. The effects of different concentrations of glycerol and dimethyl sulphoxide on the recovery of virulent and avirulent strains of Leptospira canicola and Leptospira icterohaemorrhagiae were investigated. Concentrations of cryopreservant above 5% increased the time taken for the leptospires to grow after recovery from vials stored in liquid nitrogen. In addition, the virulence of five challenge strains were shown to be little affected after 18 passages in vitro. PMID- 10799053 TI - Resistance of vaccinia virus to inactivation by solvent/detergent treatment of blood products. AB - The inactivation of enveloped viruses by two different solvent/detergent combinations, i.e. tri-n-butyl phosphate (TNBP)/Triton X-100 or TNBP/Tween 80, has been investigated using a high purity factor VIII (Replenate) and factor IX (Replenine) respectively. Treatment with TNBP/Triton X-100 rapidly inactivated all the typical enveloped viruses tested, i.e. Sindbis, semliki forest virus (SFV), herpes simplex virus type-1 (HSV-1) and vesicular stomatitis virus (VSV), by 3.7-5.8 log within 15 seconds. While virus inactivation with TNBP/Tween 80 was slower, effective inactivation of Sindbis, HSV-1, VSV and human immunodeficiency virus type-1, i.e. 4.1-->6.3 log, occurred within 30 minutes. In contrast, vaccinia virus was relatively resistant to inactivation in either of these solvent/detergent combinations. Incubation times of 10 minutes for TNBP/Triton X 100 or 6-24 hours for TNBP/Tween 80, were required to reach inactivation levels of about 4 log. PMID- 10799054 TI - In vitro potency assay for yellow fever vaccines: comparison of three vero cell lines sources. AB - Quality control of Yellow Fever vaccines performed by Control Authorities prior to marketing vaccines batches requires in vitro potency assays. The two currently available methods are the plaque formation assay and the cytopathic effect assay based on the use of porcine kidney PS cells or monkey kidney Vero cells. Among several sources of variation in virus titration, the cell systems are considered as important issues and Quality Assurance strongly recommends working with cell banks from certified suppliers. The aim of our study was to compare the behaviour and the sensitivity of three Vero cell sources obtained from ATCC, WHO and EP used at different passage levels in a plaque formation test. The conclusion of this work was that the yellow fever live attenuated virus titration, adapted in Vero cell lines appeared as a reliable method applicable for routine in vitro potency assay. The comparison of Vero cell lines, originated from three different sources, showed that they could be equally used as substrates by laboratories having the basic facility of cell culture, without influence on the final viral titre. PMID- 10799055 TI - Evaluation of vaccines, interferons and cell substrates for pestivirus contamination. AB - Pestiviruses are potential contaminants of biological products produced in bovine or porcine cells or manufactured via processes using animal-derived raw materials such as bovine serum. In order to investigate possible contamination of products including those manufactured and/or licensed in the US, 38 lots of viral vaccines and five lots of interferon alpha (IFNalpha) were tested by reverse transcriptase polymerase chain reaction (RT-PCR) for the presence of bovine viral diarrhoea virus (BVDV). All vaccines and interferons were negative for contaminating BVDV RNA when tested by RT-PCR, with the exception of an experimental live viral vaccine that had been produced in BVDV contaminated rabbit kidney cells. Cell lines commonly used to produce biological products and vaccines were experimentally infected with the NADL strain of BVDV to determine if they were permissive for virus replication. MRC-5 and WI-38 cells were not infected. In contrast, Vero, CHO and CEF cells showed evidence of pestivirus infection. Taken together these data suggested that currently licensed viral vaccines were unlikely to be contaminated with pestiviruses. However, cell banks derived from non-human primate, hamster or rabbit kidney cell lines, or cultures of primary chick embryo fibroblasts, may be infected with BVDV if exposed to pestivirus contaminated raw materials during manufacture. PMID- 10799056 TI - Biological standardization and control at the World Health Organization: successes, current issues and future challenges. PMID- 10799057 TI - The main problem with biological products of animal origin. PMID- 10799078 TI - Admitting physical examinations: should they be generic or problem-based? PMID- 10799079 TI - Documentation of rectal examination performance in the clinical teaching unit of a university hospital. AB - Digital rectal examination is used to evaluate the distal rectum and other organs, including the prostate gland. It may be combined with fecal screening for occult blood loss, and annual performance has been recommended for asymptomatic individuals over age 40 years for cancer screening. In this study, documentation of digital rectal examinations was assessed through a review of hospital medical records of a randomly selected group of 100 patient discharges (55 females and 45 males) from a total of 896 patients admitted through a hospital emergency room to a medical clinical teaching inpatient unit of a university hospital during a six month period. In this group, 26% were admitted for a gastrointestinal disorder, but only 17% of all hospitalized patients had rectal examinations done by the medical resident house staff and/or attending medical staff directly responsible for the care of these patients. Occult blood testing was done in 15 patients. Pelvic and breast examinations were rarely documented. The majority of rectal examinations (ie, 13 of 17) were 'same sex' examinations, appeared to be used largely for testing or confirmation of grossly visible blood loss and were never confirmed by attending staff. The presence or absence of nursing staff during examinations was not documented. The prostate examination was normal in one patient but not documented in the other 44 males (ie, 26 patients over age 60 years). In conclusion, rectal examinations (as well as breast and pelvic examinations) were rarely documented in the medical teaching unit by medical resident house staff or their attending staff. PMID- 10799080 TI - Effect of acid-suppressive therapy on Helicobacter pylori production of interleukin-8 in the gastric mucosa. AB - BACKGROUND: Recent studies have shown that acid-suppressive therapy increases the severity of Helicobacter pylori- associated gastritis in the corpus. PURPOSE: To evaluate interleukin (IL)-8 production in the gastric corpus mucosa before and during acid-suppressive therapy in H pylori-infected patients. PATIENTS AND METHODS: Ten patients with reflux esophagitis (five H pylori-positive and five H pylori-negative) were treated with omeprazole 20 mg. Serum gastrin concentrations, H pylori colonization density and mucosal IL-8 levels in the corpus were investigated at entry and two weeks after starting therapy. IL-8 levels were measured by ELISA. The organism density was determined, and scored according to area occupied by the bacterial colonies. The presence of H pylori was assessed by rapid urease testing and histological finding of gastric biopsy specimens. RESULTS: In H pylori-positive patients, concentrations of IL-8 during therapy significantly exceeded those before therapy (36.2+/-6. 8 versus 18.3+/ 3.8 pg/mg protein; P<0.05) without altering H pylori density. In H pylori negative patients, IL-8 levels were similar before and during therapy (6.1+/-2.7 versus 6.3+/-3.0 pg/mg protein). Concentrations of gastrin during therapy were significantly higher than those before therapy in all patients. CONCLUSIONS: The results suggest that acid suppression increases mucosal IL-8 levels in H pylori infected patients with reflux esophagitis. PMID- 10799081 TI - Applications of recombinant DNA technology in gastrointestinal medicine and hepatology: basic paradigms of molecular cell biology. Part B: eukaryotic gene transcription and post-transcriptional RNA processing. AB - The transcription of DNA into RNA is the primary level at which gene expression is controlled in eukaryotic cells. Eukaryotic gene transcription involves several different RNA polymerases that interact with a host of transcription factors to initiate transcription. Genes that encode proteins are transcribed into messenger RNA (mRNA) by RNA polymerase II. Ribosomal RNAs (rRNAs) and transfer RNAs (tRNAs) are transcribed by RNA polymerase I and III, respectively. The production of each mRNA in human cells involves complex interactions of proteins (ie, trans-acting factors) with specific sequences on the DNA (ie, cis-acting elements). Cis-acting elements are short base sequences adjacent to or within a particular gene. While the regulation of transcription is a pivotal step in the control of gene expression, a variety of molecular events, collectively known as 'RNA processing' add an additional level of control of gene expression in eukaryotic cells. (Pour le resume, voir page suivante) PMID- 10799082 TI - Lessons to be learned for gastroenterology from recent issues in clinical trial methodology. AB - Randomized trials are the preferred tool for patient-oriented research, and their main role is to enable the transfer of results from basic research to routine application. While the need for randomized trials is evident, conducting these trials is becoming increasingly difficult and complex. This article reviews actual and conflicting issues of clinical trials with respect to gastroenterology. Major problems in trial design are neglect of previous research, inadequate sample size calculations and irrelevant outcome criteria. Significant trial management problems include subversion of random allocation, and the design of systems and procedures that are inefficient, ineffective and inflexible. One of the major challenges in conducting randomized, controlled trials is obtaining informed consent because of the differing perspectives and languages of physicians and patients. Recommendations include practical guidance in obtaining informed consent, feedback of trial results to patients and support of research related to obtaining informed consent. Despite statistical guidance, several critical issues persist with respect to trial analysis. The use of confidence intervals is under-represented, the presentation of baseline data is often omitted and postsubgroup analysis is performed. Another controversial but relevant issue is the intention-to-treat analysis. Despite the formulation of standards, there is consistently poor quality of trial reporting, poor registration of unpublished trials and limited registration of ongoing trials. The authors conclude that there is a need for more randomized trials in gastroenterology. While the complexity of trial conduction has increased, so have the means of methodological and practical support. Thus, all problems can be professionally tackled, resulting in good clinical research. PMID- 10799083 TI - Do NSAIDs prevent colorectal cancer? AB - There is increasing evidence to suggest that acetylsalicylic acid (ASA) and other nonsteroidal anti-inflammatory drugs (NSAIDs) reduce the risk of colorectal cancer. This observation is supported by animal studies that show fewer tumours per animal and fewer animals with tumours after administration of several different NSAIDs. Studies in humans consistently support this hypothesis. Intervention data from familial adenomatosis coli establish that the process of human colonic adenoma polyp formation is affected. Supportive evidence comes from 21 of 23 human studies - both case-control and cohort. The reduced risk has been found in men and women, for cancers of the colon and the rectum and for the use of both ASA and the other NSAIDs. Earlier detection of lesions as a result of drug-induced bleeding does not seem to account for these findings. The molecular mechanisms responsible for the chemopreventive action of this class of drugs is not completely established. Protection may affect several pathways, including cell cycle arrest and induction of apoptosis. Because of the consistency of epidemiological, clinical and experimental data, there is no need for further placebo trials. At the same time, there is a need to establish the dose, duration and frequency of use required for cancer-preventive activity. PMID- 10799084 TI - Primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by fibrosing inflammation and obliteration of intra- and/or extrahepatic bile ducts. The disease is one of the most common cholestatic diseases in adults and is diagnosed with increasing frequency. It is very often associated with ulcerative colitis. Patients with PSC have an increased incidence of bile duct carcinomas, and those with ulcerative colitis also have an increased incidence of colonic carcinomas. In end-stage disease, liver transplantation is the treatment of choice. Immunosuppressive treatment has little effect. Ursodeoxycholic acid (UDCA), which has been shown to improve liver histology and survival in patients with primary biliary cirrhosis, has a beneficial effect in PSC, provided that patients who develop major duct stenoses are treated endoscopically. The aim is to treat patients as early as possible to prevent progression to the advanced stages of the disease. During treatment with UDCA, stenoses of major ducts may develop, and early endoscopic dilation is highly effective. Because UDCA treatment improves but does not cure cholestatic liver diseases, permanent treatment seems to be necessary. Such prolonged treatment with UDCA may be recommended because, until now, no side effects have been reported. In patients with end-stage disease, UDCA is not effective and liver transplantation is indicated. PMID- 10799085 TI - Nonalcoholic steatohepatitis: an evolving diagnosis. AB - Nonalcoholic steatohepatitis (NASH) is a histological diagnosis applied to a constellation of liver biopsy findings that develop in the absence of alcohol abuse. Steatosis, a mixed cellular inflammatory infiltrate across the lobule, evidence of hepatocyte injury and fibrosis are the findings that can be seen. This entity is often identified during evaluation of elevated aminotransferases after exclusion of viral, metabolic and other causes of liver disease. Obesity is a major risk factor for NASH. The role of diabetes is less certain, although evidence is accumulating that hyperinsulinism may play an important pathophysiological role. Patients sometimes suffer from right upper quadrant abdominal pain and fatigue; examination may reveal centripetal obesity and hepatomegaly. Although patients are often discovered because of persistent aminotransferase elevations, these enzymes can be normal in NASH. When they are elevated, the alanine aminotransferase level is typically significantly greater than the aspartate aminotransferase level. This can be particularly helpful for excluding occult alcohol abuse. Imaging studies identify hepatic steatosis when the amount of fat in the liver is significant; however, imaging does not distinguish benign steatosis from NASH. Ultimately a liver biopsy is needed to diagnose NASH. The biopsy may be useful for establishing prognosis based on the presence or absence of fibrosis and for excluding other unexpected causes of liver enzyme elevations. Weight loss is the mainstay of treatment for obese patients. About 15% to 40% of NASH patients develop fibrosis; how many of these cases progress to cirrhosis is unknown, but about 1% of liver transplants are performed with a pretransplant diagnosis of NASH. PMID- 10799086 TI - Mechanisms of alcoholic liver injury. AB - There have been numerous recent advances in the understanding of the mechanisms of alcoholic liver disease pathogenesis. Endotoxin-induced Kupffer cell activation plays a role in cytokine-mediated inflammatory changes in the liver, and this can be blocked by a diet high in saturated fat, by a diet containing lactobacillus, which does not produce endotoxin, by neomycin antibiotic sterilization of the gut, by eliminating Kupffer cells, or by removing tumour necrosis factor-alpha with antibody or by using tumour necrosis factor-alpha knockout mice. The fatty liver component is mainly the result of the nicotinamide adenine dinucleotide/reduced nicotinamide adenine dinucleotide redox shift to the reduced state by ethanol oxidation generation of reduced nicotinamide adenine dinucleotide, although this too can be blocked by a diet high in saturated fat. Hepatocytic enlargement occurs due to ethanol-induced inhibition of the ubiquitin proteasome pathway of cytoplasmic protein degradation and the retention of oxidized proteins in hepatocytes. The liver is scarred by stellate cells that have been activated by inflammatory cytokines and growth factors produced by activated Kupffer cells, and by bile ductule metaplasia. Mallory bodies and balloon cell degeneration develop through the ethanol-induced oxidative stress protein kinase activation pathway, inhibition of phosphatase activity and inhibition of the ubiquitin-proteasome pathway. PMID- 10799087 TI - Hepatitis in disseminated bacillus Calmette-Guerin infection. AB - Local immunotherapy with an attenuated live strain of Mycobacterium bovis, bacillus Calmette-Guerin (BCG), is an effective and frequently used treatment for in situ transitional cell carcinoma (TCC) of the bladder. Success rates are high, and serious side effects are infrequent but can affect every organ system. A 79 year-old patient with recently diagnosed TCC who was treated with intravesical BCG for a recurrence after initial surgical treatment is reported. After unsuccessful attempts at bladder catheterization with the creation of a false passage for his third treatment, BCG was instilled via a suprapubic catheter the same day and again a week later. Two weeks after the third BCG instillation, the patient presented with profound lethargy and weakness to the point of not being able to get up out of a chair. He was febrile, anorexic, icteric and had hepatosplenomegaly. Disseminated BCG infection was suspected on the basis of history, clinical examination and a liver biopsy that showed noncaseating granulomatous hepatitis. Empirical treatment was started with antituberculous combination therapy. A short course of an oral corticosteroid was given. Clinical improvement was marked and sustained so that the patient could be discharged home for the full six-month course of his treatment. Disseminated BCG infection with granulomatous hepatitis can be severe and life-threatening in cases where a large intravascular inoculum of BCG may have been given inadvertently. PMID- 10799088 TI - Hereditary angioneurotic edema and familial Crohn's disease. AB - A 29-year-old man with Crohn's disease involving the ileum and cecum was seen. He had angioneurotic edema with C1' esterase inhibitor deficiency. Later, his 50 year-old mother was evaluated because of abdominal pain. She had recurrent urticaria, C1' esterase inhibitor deficiency and radiographic studies showed Crohn's disease of the ileum. A maternal family history revealed other members affected with either Crohn's disease or angioneurotic edema. The clinical observations in this family suggest that angioneurotic edema associated with C1' esterase inhibitor deficiency may be closely linked genetically with a familial form of Crohn's disease. PMID- 10799089 TI - An unusual cause of recurrent pancreatitis: duodenal duplication cyst. AB - Intestinal duplications are rare anomalies that usually present in childhood. Of these, duodenal duplications are among the most uncommon. Although these usually present with obstructive symptoms or bleeding, pancreatitis may occur. A case of duodenal duplication cyst causing recurrent pancreatitis in a 12-year-old girl is presented. The literature on duodenal duplications, and their epidemiology, embryology and pathophysiology is reviewed. PMID- 10799090 TI - C1 inhibitor deficiency and angioedema of the small intestine masquerading as Crohn's disease. AB - A case of C1 inhibitor deficiency presenting as localized edema of the small intestine is described. A 16-year-old, previously healthy woman presented with recurrent attacks of abdominal pain and vomiting following minor abdominal trauma. Investigations including computed tomography scan and barium studies confirmed localized edema of the jejunum. At laparoscopy, Crohn's disease was suspected; however, a subsequent enteroscopy was normal. Complement levels revealed a low C4 level, and C1 inhibitor deficiency was later confirmed. Attacks of abdominal pain began after starting oral contraceptives and have not returned since stopping the birth control pill. This rare cause of abdominal pain is examined, and C1 inhibitor deficiency and angioedema are reviewed. PMID- 10799091 TI - Synchronous occurrence of collagenous colitis and pseudomembranous colitis. AB - Synchronous collagenous and pseudomembranous colitis has not been previously reported. A 73-year-old woman presented with chronic watery diarrhea and abdominal cramping of six weeks' duration. Biopsies of the colon revealed findings of collagenous colitis involving the endoscopically normal right colon, and superimposed collagenous and pseudomembranous colitis involving the rectosigmoid colon. Endoscopically, the left colon revealed discrete ulcerative plaques, and Clostridium difficile toxin A assay was positive. The patient partially responded to a three-week regimen of metronidazole, and symptoms resolved completely with subsequent steroid therapy. At follow-up endoscopy four months later, colon biopsies demonstrated persistence of subepithelial collagen but no pseudomembranes. The patient remained asymptomatic during this interval. Collagenous colitis has been reported in association with other inflammatory bowel diseases, including lymphocytic colitis, sprue and idiopathic inflammatory bowel disease. This unique association of collagenous colitis with an endotoxigenic inflammatory bowel disease is presented with a review of related disease features. PMID- 10799092 TI - Patellar resurfacing in total knee arthroplasty. AB - Whether or not to resurface the patella when performing a primary total knee arthroplasty remains an open question. A number of recent studies have added new information relevant to this controversy. Anatomic studies show that there is normally substantial variability in the anatomy of the trochlear groove. Implanting a femoral component therefore results in a change in the surface topography of the knee in a high percentage of cases. Even though a number of intraoperative techniques have been described in an attempt to accurately reproduce femoral and tibial component rotation, studies of the application of these techniques reveal that component malpositioning or malrotation of a measurable degree occurs in 10% to 30% of cases, depending on the surgical technique and landmarks used. There has been substantial change in the design of both femoral and patellar components in recent years. Even with current designs, biomechanical studies indicate that some degree of change in kinematics and contact stresses occurs following total knee arthroplasty. However, the results of clinical studies have been extremely variable, with most showing either no difference or very little difference between resurfaced and nonresurfaced patellae in osteoarthritic knees. The decision to resurface the patella or not must be individualized on the basis of the surgeon's training and experience and an intraoperative assessment of the patellofemoral articulation. PMID- 10799093 TI - Intramedullary nailing of the femur: reamed versus nonreamed. AB - All intramedullary nailing creates some loss of endosteal blood supply and an increase in intramedullary pressure, resulting in marrow embolization. In laboratory studies, both reamed and nonreamed intramedullary nailing have led to alteration in selected pulmonary variables. This effect, although transient, appeared more pronounced with reamed techniques than with nonreamed techniques. Concern about the systemic pulmonary effects of reamed intramedullary nailing has led to an increase in the use of nonreamed nailing. The authors of most clinical studies have reported that reamed intramedullary nailing has not been associated with a concomitant increase in pulmonary complications in multiply injured patients, although this point is still controversial. Femoral shaft fractures treated with nonreamed nailing have been shown to have slightly higher rates of delayed union and nonunion compared with those treated with reamed nails. Reamed interlocking intramedullary fixation remains the treatment of choice for femoral shaft fractures in adults. Further study is required to determine whether an identifiable subgroup of trauma patients is adversely affected by intramedullary reaming, which would suggest the need for alternative fixation techniques. PMID- 10799094 TI - Domestic violence: the role of the orthopaedic surgeon in identification and treatment. AB - Domestic violence is a major public health problem in the United States. As many as 35% of women visiting hospital emergency departments for trauma care are there because of injuries caused by intimate partner violence. The practicing orthopaedic surgeon may come in contact with these women in the emergency department as well as in the office setting. The ability to identify victims of abuse requires a sensitive approach and a specific set of skills. Once the victim has been identified, appropriate referral to local agencies is critical to help ensure the victim's safety. The issues surrounding identification, documentation, inquiry about safety, and activation of community services need to be incorporated into the core curriculum of resident training programs and the continuing education of the practicing orthopaedic surgeon. PMID- 10799095 TI - Acute and chronic posterolateral rotatory instability of the knee. AB - Isolated posterolateral rotatory instability of the knee is an uncommon injury pattern that may result in significant degrees of functional disability. This injury complex can be a challenging diagnostic and therapeutic problem for the orthopaedic surgeon. The presence of associated ligamentous and soft-tissue injuries, resulting in combined instability patterns, further complicates management. The results of recent research have enhanced our understanding of the complex anatomy and biomechanics of the posterolateral aspect of the knee. Numerous surgical techniques have been described for both repair and reconstruction of the injured posterolateral structures; however, long-term functional results have been only moderately successful. PMID- 10799096 TI - Operative treatment of metacarpal and phalangeal shaft fractures. AB - Diaphyseal fractures of the metacarpals and phalanges are common injuries that can lead to impairment of hand function. The fracture pattern and soft-tissue injury vary with the mechanism of injury. The imbalance of the flexor and extensor forces created by displaced fractures will often produce a secondary angulatory deformity. Nonoperative treatment is indicated for reducible and stable fracture configurations. Irreducible or unstable fracture patterns require open or closed reduction and fixation. Reduction must be assessed in flexion and extension to ensure correct rotatory alignment. Fracture fixation can be achieved with the use of Kirschner wires, interfragmentary screws, or plates. The outcome after surgery is greatly influenced by the condition of the surrounding soft tissues; therefore, surgical trauma should be minimized to optimize the result. PMID- 10799097 TI - Complications of open anterior stabilization of the shoulder. AB - Complications of surgery for glenohumeral instability are relatively uncommon. When they occur, salvaging failures and obtaining a stable joint can be awesome challenges. Accurate recognition of the cause of the instability and application of the appropriate surgical technique are critical. Deficiencies of the glenoid concavity, the anterior capsule, or the subscapularis may be present and require correction. Overtightening a shoulder and eliminating its normal laxity should be avoided. Loose or malpositioned hardware about the glenohumeral joint must be recognized as soon as possible and removed. The goal of treatment is to correct the deficient stabilizing mechanisms without altering normal glenohumeral function. PMID- 10799099 TI - Orthodontia: its outlook. PMID- 10799098 TI - Quality and outcome determination in health care and orthopaedics: evolution and current structure. AB - Quality health care has many definitions. Among those definitions is "care that consistently contributes to the improvement or maintenance of the quality and/or duration of life." The current evolution in health care has been fueled by three necessities frequently demanded by payers and employers: improvement in access, lowering of cost, and definition and quantification of the quality of care. This evolution has been facilitated by the so-called industrialization of medicine. This concept includes the adoption of industrial economic principles and techniques that facilitate the measurement of processes and outcomes. Quality health care is currently recognized as health care that is characterized by three elements: the use of practice guidelines or standards, the implementation of continuous quality improvement techniques, and the use of outcome determination and management. Practice guidelines demand the adoption of evidence-based principles in evaluation and care, as well as minimization of variations in evaluation and care. Continuous quality improvement seeks to determine why variations in processes of care occur and then to minimize those variations. Outcomes may be measured in terms of both very objective and very subjective variables and also on the basis of cost-efficiency. Most tools currently used to quantify outcomes, especially in orthopaedics, involve measurements of general health and of specific body part or organ system function. This evolution in health care is producing significant alterations in methods of traditional health care delivery. The accumulating evidence indicates that these changes, although frequently unpopular, are improving the quality of health care. PMID- 10799100 TI - Vignette: T. M. Graber. PMID- 10799101 TI - The American Association of Orthodontists at 2000: some thoughts for the new millennium. PMID- 10799102 TI - The American Association of Orthodontists: for our common good, today and tomorrow. PMID- 10799103 TI - American Board of Orthodontics: past, present, and future. PMID- 10799104 TI - American Association of Orthodontists Foundation: past, present, and future. PMID- 10799105 TI - The World Federation of Orthodontists: "bringing the world together". PMID- 10799106 TI - Toward understanding the molecular basis of craniofacial growth and development. PMID- 10799107 TI - Stop me before I write again .... PMID- 10799108 TI - Evidence-based treatment strategies: an ambition for the future. PMID- 10799109 TI - The evolution of orthodontics to a data-based specialty. PMID- 10799110 TI - Clinical research about clinical treatment: a new agenda for a new century. PMID- 10799111 TI - Tomorrow's challenges for the science of orthodontics. PMID- 10799113 TI - A statement regarding early treatment. PMID- 10799112 TI - The contributions of craniofacial growth to clinical orthodontics. PMID- 10799114 TI - The significance of late developmental crowding to early treatment planning for incisor crowding. PMID- 10799115 TI - Orthodontic relapse versus natural development. PMID- 10799116 TI - Serial extraction ... nobody does that anymore! PMID- 10799117 TI - Maxillary transverse deficiency. PMID- 10799118 TI - Dentofacial orthopedics or orthognathic surgery: is it a matter of age? PMID- 10799119 TI - Orthodontics about face: the re-emergence of the esthetic paradigm. PMID- 10799120 TI - Mysteries of asymmetries. PMID- 10799121 TI - 2D or not 2D? That is the question. PMID- 10799122 TI - Looking back and forward through my career in orthodontics. PMID- 10799123 TI - Orthodontic magic. PMID- 10799124 TI - Clubs, quips, phrases, and hype: musings for the new millennium. PMID- 10799125 TI - Orthodontic biomechanics: vistas from the top of a new century. PMID- 10799126 TI - Orthodontic bonding to artificial tooth surfaces: clinical versus laboratory findings. PMID- 10799127 TI - Ceramic brackets and the need to develop national standards. PMID- 10799128 TI - Orthodontics as a science: the role of biomechanics. PMID- 10799129 TI - Enhancing the value of orthodontic treatment: incorporating effective preventive dentistry into treatment. PMID- 10799130 TI - The evolutionary tidal wave. PMID- 10799132 TI - One viewpoint on teaching clinical orthodontics. PMID- 10799131 TI - Contemporary technology-centered practice. PMID- 10799133 TI - The winds of change. PMID- 10799134 TI - Orthodontics in the next 100 years: prediction or speculation? PMID- 10799135 TI - The decade ahead: finding a better way. PMID- 10799136 TI - Pride in orthodontics. PMID- 10799137 TI - William A. Mitchell, jr, april 26, 1933 to march 17, 2000 PMID- 10799138 TI - Do you have a satellite office in cyberspace? PMID- 10799139 TI - Litigation, legislation, and ethics. If a professional practice is a small business .... PMID- 10799140 TI - Women and heart disease. PMID- 10799141 TI - Was the danger of diet pills overstated? PMID- 10799142 TI - Hidden benefits of ACE inhibitors. PMID- 10799143 TI - Vigorous activity lowers diabetes risk. PMID- 10799144 TI - Ask the doctor: My heart was damaged by two heart attacks. I know that ACE inhibitors help patients with heart failure, but I just can't take them. They make me cough. is there something else I can try? PMID- 10799145 TI - New twist on the old stethoscope. PMID- 10799146 TI - Beta blockers safe and helpful for diabetics with heart attacks. PMID- 10799147 TI - A gooey but effective cholesterol medication. PMID- 10799148 TI - Exhaustion and heart disease. PMID- 10799149 TI - Low cholesterol doesn't make smoking safer. PMID- 10799150 TI - In the pipeline... PMID- 10799151 TI - Prostate specific antigen only progression of prostate cancer. AB - PURPOSE: Introduction of the prostate specific antigen (PSA) serum marker for prostate cancer and the subsequent PSA era from 1988 to the present have dramatically altered the diagnosis of the disease. The early to mid 1990s diagnosis boom resulted in a huge increase in clinically localized and early stage disease treatments. Radical prostatectomy rates increased from 17.4 to 54.6/100,000 between 1988 and 1992, and age adjusted rates increased 2 to 4-fold for men in the fifth and sixth decades of life. Since the late 1990s clinicians have been seeing the effects of this diagnosis and localized treatment boom, in that many men each year are experiencing PSA only disease recurrence. Given that the majority are relatively young and otherwise healthy, treatment of PSA only recurrence requires approaches that not only improve survival, but also preserve quality of life. A comprehensive overview of the definition of PSA only recurrence, staging controversies and the wide variety of treatments to be considered is provided. MATERIALS AND METHODS: A literature review and overview of the topic of PSA only recurrence after prior clinically localized prostate cancer treatment were performed. RESULTS: For radical prostatectomy cases PSA only recurrence is broadly defined as any elevation of PSA postoperatively. For radiation treated patients the 1997 American Society for Therapeutic Radiology and Oncology guidelines specify 3 consecutive elevations of PSA after posttreatment PSA nadir is achieved. As localized treatment series in the PSA era have matured, and database and statistical support have improved, a number of useful models to predict PSA only recurrence have emerged. These models are based on traditional prognostic markers, such as pretreatment PSA, and grade and stage of disease as well as emerging molecular and cellular biomarkers. Although bone scans and pelvic computerized tomography are commonly used for re-staging at PSA only recurrence, recent study suggests that their value is limited unless PSA recurrence exceeds 30 to 40 ng./ml. 111Indium capromab pendetide radionuclide scan, which has been approved for radical prostatectomy PSA only recurrence, may be helpful to determine cases best suited for salvage radiotherapy versus systemic hormonal therapy, although more study is needed. Treatment of PSA only recurrence is divided into 2 main categories of salvage local treatments and systemic therapy. The principal dilemma is the inability to determine definitively whether PSA only recurrence is solely due to local progression or distant micrometastases. External beam radiation is the main local salvage treatment for radical prostatectomy recurrence, and cryotherapy, salvage prostatectomy and salvage brachytherapy are options for radiation recurrence. Proper patient selection is critical to the success of all salvage local treatments. Traditional hormonal therapy is the mainstay of systemic treatment for PSA only recurrence, although nontraditional approaches, such as intermittent and low dose hormonal therapy, are under study. Emerging chemopreventive agents, such as vitamins, minerals and other supplements, may have a future role in treatment. CONCLUSIONS: PSA only recurrence after prior local prostate cancer treatment remains a common problem facing clinicians. PMID- 10799152 TI - Laparoscopic radical prostatectomy: the Montsouris technique. AB - PURPOSE: Laparoscopic radical prostatectomy has become standard at our institution based on experience with 260 consecutive cases operated on between January 1998 and December 1999. In view of the favorable short-term outcomes we describe our standardized laparoscopic radical prostatectomy technique. MATERIALS AND METHODS: Two urologists trained in open retropubic radical prostatectomy and laparoscopy combined their experience to develop a specific technique of nonincisional radical prostatectomy for localized prostate cancer. Patients presented with clinical stages T1b to T2 prostate cancer and tumor size was approximately 18 to 130 gm. Operations were performed by 1 senior surgeon and 1 assistant, with the help of a voice controlled robot and with the patient under general anesthesia. The 2, 10 mm. ports and 3, 5 mm. ports were placed in the umbilicus and iliac fossa. The laparoscopic procedure was performed transperitoneally, combining anterograde and retrograde approaches in 7 standardized steps. Urethrovesical anastomosis was performed with 3-zero interrupted sutures tied intracorporeally. Technical details were compiled, summarized and illustrated with schematic views. RESULTS: Operating time was approximately 3 hours for the last 120 cases. Estimated average blood loss was 250 ml. with a transfusion rate of less than 1%. The conversion rate was 0%. Postoperative pain was minimal and analgesics were generally not required by postoperative day 2. The accuracy of dissection and sutures allowed patients to be discharged home without urethral catheterization starting on postoperative day 3. CONCLUSIONS: Laparoscopic radical prostatectomy is now not only feasible, but more importantly reproducible. Each step has been checked and validated, and the procedure is standardized and has definitively replaced the retropubic approach in our practice. PMID- 10799153 TI - Prospective, case matched comparison of hand assisted laparoscopic and open surgical live donor nephrectomy. AB - PURPOSE: The technical difficulty of standard laparoscopic live donor nephrectomy has limited its application. Hand assistance, which takes advantage of the incision necessary for organ removal, facilitates laparoscopy without significant impact on patient recovery. We prospectively compared open surgical and hand assisted laparoscopic donor nephrectomy. MATERIALS AND METHODS: Our first 10 laparoscopic live donor nephrectomies were matched with 40 open donor nephrectomies by gender, age and body mass index. Data were obtained by pain scales, SF-12 survey instruments, questionnaires and chart abstraction. RESULTS: Operative time was longer for the laparoscopic approach (mean 95 versus 215 minutes). However, laparoscopic group patients had a shorter hospital stay compared to those undergoing open surgery (mean 2.9 versus 1.8 days), returned sooner to nonstrenuous activity (mean 19.0 versus 9.9 days) and reported less pain 6 weeks postoperatively (mean 2.3 versus 0.6) (p P2X5 = P2X6. P2X3 immunoreactivity was seen within nerve bundles in detrusor muscle only. The fine capillary network supplying bladder and ureter smooth muscle and lamina propria was visualized with P2X4 immunoreactivity, membranes of urothelial cells gave a strong reaction with P2X5, whereas P2X6 immunostained the thin basement membrane beneath the urothelium. Nuclear staining was seen with P2X7 in the urothelium but more prominent in the bladder than in the ureter. CONCLUSIONS: Having established the distribution of P2X receptors in normal animal bladder and ureter tissue, it is now possible to perform comparable investigations on normal and diseased human tissue to establish a possible role of P2X receptors in pathogenic events. PMID- 10799248 TI - Improved contractility of obstructed bladders after Tadenan treatment is associated with reversal of altered myosin isoform expression. AB - PURPOSE: Tadenan is a plant extract from Pygeum africanum used in the treatment of benign prostatic hyperplasia, to protect the bladder from contractile dysfunction induced by partial bladder outlet obstruction (BOO). The aim of the present study was to determine whether the Tadenan-induced return of detrusor contractility affects the expression of myosin isoforms, which differ at the C terminal (SM1 and SM2) and the N-terminal regions (SM-A and SM-B). MATERIALS AND METHODS: Four groups of New Zealand White rabbits (3 to 5 kg., 4 to 6 rabbits per group) were either partially obstructed by ligation of the urethra (groups 1 and 2) or not obstructed (groups 3 and 4). After 2 weeks, rabbits from groups 2 and 4 received Tadenan in peanut oil (vehicle) orally at 100 mg. /kg./day for 3 weeks and rabbits in groups 1 and 3 received vehicle only. Rabbits were sacrificed and bladders were removed and weighed. Contractility studies were performed on isolated strips of detrusor and the remaining muscular layer from the bladder body was used to study the expression of myosin heavy chain (MHC) isoforms at mRNA (SM1, SM2, SM-A, and SM-B) and the protein (SM1 and SM2) levels by RT-PCR and SDS-PAGE analyses, respectively. RESULTS: Tadenan significantly reduced the effect of BOO on bladder mass. The diminished contractile response to field stimulation and carbachol secondary to urethral obstruction was significantly reversed by Tadenan treatment. The relative ratios for MHC isoforms were altered at the mRNA (SM2:SM1 and SM-A:SM-B) and protein (SM2:SM1) levels in obstruction. Upon treatment with Tadenan, the ratio of these isoforms returned to normal, as shown at the mRNA levels. In addition, the altered relative ratio of SM2:SM1 at the protein level also returned to nearly normal values after treatment. CONCLUSIONS: Improvement of obstruction-induced contractile dysfunction of the detrusor following treatment with Tadenan is associated with changes in the expression of myosin isoforms. The alteration in the expression of myosin isoforms associated with obstruction-induced hypertrophy is reversed close to normal in the detrusor smooth muscle from Tadenan-treated obstructed rabbits. PMID- 10799249 TI - BCG-induced urinary cytokines inhibit microvascular endothelial cell proliferation. AB - PURPOSE: Angiogenesis is thought to depend on a net balance of molecules that inhibit or stimulate microvascular endothelial cells. A variety of molecules that affect angiogenesis are induced locally by the administration of intravesical bacille Calmette-Guerin (BCG) for superficial bladder cancer. We sought to determine whether BCG-induced urinary cytokines alter the effects of patient urine on assays of angiogenic activity. MATERIALS AND METHODS: Patients undergoing BCG treatment provided urine samples before and at peak cytokine production times after BCG instillation. Fifty-four urine samples from 8 patients were analyzed by ELISA for a panel of molecules known to affect angiogenesis, and tested for angiogenic activity in human dermal microvascular endothelial cell (HDMEC) proliferation and migration assays. To assess the role of specific BCG induced cytokines, urinary HDMEC proliferation assays were repeated in the presence of neutralizing antibodies to tumor necrosis factor-alpha (TNF-alpha), interferon-inducible protein-10 (IP-10), and/or interferon-gamma (IFN-gamma). RESULTS: Urinary IFN-gamma, IP-10, TNF-alpha, and vascular endothelial growth factor (VEGF) were induced to nanogram/ml amounts by BCG treatment. While pre-BCG treatment urine samples minimally stimulated microvascular endothelial cell proliferation (+ 9%), post-BCG treatment urine became progressively inhibitory to endothelial cells (to -85%, p = 0.005) during weekly treatment courses. Neutralizing antibodies to TNF-alpha or to IP-10, either alone or in combination, greatly reduced this inhibitory effect. CONCLUSIONS: Intravesical BCG induces a cytokine-rich urinary microenvironment that is inhibitory to human endothelial cells. Urinary cytokine profiles and assays of angiogenic inhibition may provide prognostically important information regarding BCG treatment outcomes. PMID- 10799251 TI - Ask the doctor. I had a heart attack last year and have resumed most of my activities, including some tennis, without problems. However, both my wife and I are reluctant to have sexual intercourse. To be frank, she is concerned that it would be dangerous for me. Is she right? If not, what can I do to put her mind at ease? PMID- 10799250 TI - Detection of circulating prostate derived cells in patients with prostate adenocarcinoma is an independent risk factor for tumor recurrence. AB - PURPOSE: To determine whether the presence of prostate-derived cells in the peripheral blood circulation is a marker of prostate cancer and to define the clinical impact of the test. MATERIALS AND METHODS: We tested the peripheral blood of 99 patients with prostate adenocarcinoma (PAC), 79 of them undergoing radical prostatectomy, and 92 controls (31 healthy volunteers, 50 patients with adenoma and 11 with prostatitis) using a highly controlled procedure including reverse-transcriptase polymerase chain reaction (RT-PCR) targeted to prostate specific antigen (PSA) mRNA. Patients were followed for 26 +/- 12 (range: 4 to 49) months. Forty tumor tissues were analyzed by immunohistochemistry for expression of p53 and E-cadherin antigens. RESULTS: Thirty three (33%) patients with PAC and 2 (2%) controls scored positive (p <0.0001) for the test. Detection of circulating prostatic cells was associated with development of metastases (p <0. 001), with relapse (p <0.001) and with a serum PSA level at diagnosis higher than 15 ng./ml. (p = 0.009). The rate of development of metastases according to time was significantly higher in patients who scored positive for the test (p <0.04). In a multivariate analysis, only the RT-PCR test was an independent risk factor associated with relapse (RR: 6.7). Finally, E-cadherin expression was significantly lower in the tumor tissues of positive patients as compared with those who scored negative for the test (p <0.01). CONCLUSIONS: This RT-PCR procedure, performed at diagnosis and with appropriate controls, is a clinically useful assay in evaluating the risk of tumor recurrence after radical prostatectomy in patients with PAC. PMID- 10799252 TI - Heart effects of herbal medicine. PMID- 10799254 TI - People with high blood pressure more prone to panic disorder. PMID- 10799253 TI - Endoscopic vein harvesting. PMID- 10799255 TI - Ask the doctor. I take atenolol and lisinopril for my blood pressure. I have a blood pressure monitor that I use at home and I find that, on average, my pressure is 150/85 in the morning and 130/80 in the afternoon. Should I be worried about the high morning readings? PMID- 10799256 TI - New skeletal remains of Omomys (Primates, Omomyidae): functional morphology of the hindlimb and locomotor behavior of a Middle Eocene primate. AB - New associated craniodental and postcranial remains of Omomys carteri from Bridger C beds in Uinta County, Wyoming represent the largest and most nearly complete single taxon sample of omomyid postcranial fossils presently known. They allow, for the first time, a description and detailed analysis of the functional morphology of the hindlimb of this middle Eocene omomyine primate. Comparisons of pelvic, femoral, tibial, and pedal morphology and metrics with a large sample of prosimian primates of known locomotor behavior suggest that Omomys possessed a highly distinctive mosaic of functional adaptations related to active quadrupedalism and leaping. Traits suggestive of quadrupedalism include the lengths of the ischium and ilium, position of the femoral trochanters, and lengths and features of the tarsal bones. Morphological traits that suggest leaping include a semi-cylindrical femoral head with moderate posterior expansion of the articular surface, greater trochanter projecting anterior to the femoral shaft, deeper than wide femoral condyles, narrow and deep patellar groove with prominent lateral border, elongated calcaneus, and close aposition of distal tibia and fibula. While Omomys most closely resembles active quadrupedal cheirogaleids like Cheirogaleus and Mirza, leaping must have been an important component of its locomotor repertoire. In this respect, Omomys closely resembles other North American omomyines (notably Hemiacodon), and is significantly more generalized postcranially than European microchoerines (e.g., Microchoerus, Nannopithex, and Necrolemur). PMID- 10799257 TI - New wrist bones of the Malagasy giant subfossil lemurs. AB - Recently discovered wrist bones of the Malagasy subfossil lemurs Babakotia radofilai, Palaeopropithecus ingens, Mesopropithecus dolichobrachion, and Megaladapis madagascariensis shed new light on the postcranial morphologies and positional behaviors that characterized these extinct primates. Wrist bones of P. ingens resemble those of certain modern hominoids in having a relatively enlarged ulnar head and dorsally extended articular surface on the hamate, features related to a large range of rotation at the inferior radioulnar and midcarpal joints. The scaphoid of P. ingens is also similar to that of the extant tree sloth Choloepus in having an elongate, palmarly directed tubercle forming a deep radial margin of the carpal tunnel for the passage of large digital flexors. In contrast, wrist remains of Megaladapis edwardsi and M. madagascariensis exhibit traits observed in the hands of extant pronograde, arboreal primates; these include a dorsopalmarly expanded pisiform and well-developed "spiral" facet on the hamate. Moreover, Megaladapis spp. and Mesopropithecus dolichobrachion possess bony tubercles (e.g., scaphoid tubercle and hamate hamulus) forming the carpal tunnel that are relatively similar in length to those of modern pronograde lemurs. Babakotia and Mesopropithecus differ from Megaladapis in exhibiting features of the midcarpal joint related to frequent supination and radioulnar deviation of the hand characteristic of animals that use vertical and quadrumanous climbing in their foraging behaviors. Comparative analysis of subfossil lemur wrist morphology complements and expands upon prior inferences based on other regions of the postcranial skeleton, and suggests a considerable degree of locomotor and postural heterogeneity among these recently extinct primates. PMID- 10799258 TI - Locomotor energetics and leg length in hominid bipedality. AB - Because bipedality is the quintessential characteristic of Hominidae, researchers have compared ancient forms of bipedality with modern human gait since the first clear evidence of bipedal australopithecines was unearthed over 70 years ago. Several researchers have suggested that the australopithecine form of bipedality was transitional between the quadrupedality of the African apes and modern human bipedality and, consequently, inefficient. Other researchers have maintained that australopithecine bipedality was identical to that of Homo. But is it reasonable to require that all forms of hominid bipedality must be the same in order to be optimized? Most attempts to evaluate the locomotor effectiveness of the australopithecines have, unfortunately, assumed that the locomotor anatomy of modern humans is the exemplar of consummate bipedality. Modern human anatomy is, however, the product of selective pressures present in the particular milieu in which Homo arose and it is not necessarily the only, or even the most efficient, bipedal solution possible. In this report, we investigate the locomotion of Australopithecus afarensis, as represented by AL 288-1, using standard mechanical analyses. The osteological anatomy of AL 288-1 and movement profiles derived from modern humans are applied to a dynamic model of a biped, which predicts the mechanical power required by AL 288-1 to walk at various velocities. This same procedure is used with the anatomy of a composite modern woman and a comparison made. We find that AL 288-1 expends less energy than the composite woman when locomoting at walking speeds. This energetic advantage comes, however, at a price: the preferred transition speed (from a walk to a run) of AL 288-1 was lower than that of the composite woman. Consequently, the maximum daily range of AL 288-1 may well have been substantially smaller than that of modern people. The locomotor anatomy of A. afarensis may have been optimized for a particular ecological niche-slow speed foraging-and is neither compromised nor transitional. PMID- 10799259 TI - Masticatory stress, orbital orientation and the evolution of the primate postorbital bar. AB - A postorbital bar is one of a suite of derived features which distinguishes basal primates from their putative sister taxon, plesiadapiforms. Two hypotheses have been put forward to explain postorbital bar development and variation in circumorbital form: the facial torsion model and visual predation hypothesis. To test the facial torsion model, we employ strain data on circumorbital and mandibular loading patterns in representative primates with a postorbital bar and masticatory apparatus similar to basal primates. To examine the visual predation hypothesis, we employ metric data on orbit orientation in Paleocene and Eocene primates, as well as several clades of visual predators and foragers that vary interspecifically in postorbital bar formation.A comparison of galago circumorbital and mandibular peak strains during powerful mastication demonstrates that circumorbital strains are quite low. This indicates that, as in anthropoids, the strepsirhine circumorbital region is excessively overbuilt for countering routine masticatory loads. The fact that circumorbital peak-strain levels are uniformly low in both primate suborders undermines any model which posits that masticatory stresses are determinants of circumorbital form, function and evolution. This is interpreted to mean that sufficient cortical bone must exist to prevent structural failure due to non-masticatory traumatic forces. Preliminary data also indicate that the difference between circumorbital and mandibular strains is greater in larger taxa.Comparative analyses of several extant analogs suggest that the postorbital bar apparently provides rigidity to the lateral orbital margins to ensure a high level of visual acuity during chewing and biting. The origin of the primate postorbital bar is linked to changes in orbital convergence and frontation at smaller sizes due to nocturnal visual predation and increased encephalization. By incorporating in vivo and fossil data, we reformulate the visual predation hypothesis of primate origins and thus offer new insights into major adaptive transformations in the primate skull. PMID- 10799260 TI - Early hominid brain evolution: a new look at old endocasts. AB - Early hominid brain morphology is reassessed from endocasts of Australopithecus africanus and three species of Paranthropus, and new endocast reconstructions and cranial capacities are reported for four key specimens from the Paranthropus clade. The brain morphology of Australopithecus africanus appears more human like than that of Paranthropus in terms of overall frontal and temporal lobe shape. These new data do not support the proposal that increased encephalization is a shared feature between Paranthropus and early Homo. Our findings are consistent with the hypothesis that Australopithecus africanus could have been ancestral to Homo, and have implications for assessing the tempo and mode of early hominid neurological and cognitive evolution. PMID- 10799261 TI - Redating Australia's oldest human remains: a sceptic's view. PMID- 10799262 TI - On the reliability of age estimates for human remains at lake mungo. PMID- 10799263 TI - Age of the Lake Mungo 3 skeleton, reply to Bowler & Magee and to Gillespie & Roberts. PMID- 10799264 TI - Australian Pleistocene variation and the sex of Lake Mungo 3. PMID- 10799265 TI - Influence of DNA polymerases on quantitative PCR results using TaqMan probe format in the LightCycler instrument. AB - Real-time fluorescence polymerase chain reaction (PCR) techniques are increasingly used to quantitate target sequences for diagnostic and research purposes. Currently, the so called TaqMan probe chemistry is mostly used as fluorogenic system. This probe format is strictly dependent on the 5'-exonuclease activity of DNA polymerase as fragmentation of the probe during the reaction is essential for this assay. Based on our experience that dramatic differences in quantitative PCR results may be due to different DNA polymerases we performed a detailed comparison of 15 enzymes. We found that clear differences exist between polymerases of different manufacturers. Thus, three out of seven polymerases which were declared to possess 5'-exonuclease activity appeared to be completely unsuitable for this method while the remaining had significantly different reaction efficiencies. We conclude that different DNA polymerases may determine the entire analytical performance of TaqMan assays suggesting that DNA polymerase testing is of special importance when this probe format is used. PMID- 10799266 TI - Rapid diagnosis of acute pyogenic meningitis by a combined PCR dot-blot assay. AB - A multiplex PCR was employed to amplify unique conserved sequences of DNA from the pathogens Haemophilus influenzae, Neisseria meningitidis and Streptococcus pneumoniae from cerebrospinal fluid samples of patients suffering from acute pyogenic meningitis. The accurate identification of the PCR amplified product was achieved by hybridizing dot-blots of the PCR products to probes which were specific, biotinylated internal sequences of the amplified target DNA. Detection of the hybrids was done in a colour reaction using streptavidin-alkaline phosphatase conjugate and BCIP/NBT substrates. The entire protocol took only 7 h for the correct identification of the pathogen present in clinical samples of cerebrospinal fluid. The sensitivity and specificity were >95%. PMID- 10799267 TI - Identification of Staphylococcus aureus enterotoxins A and B genes by PCR-ELISA. AB - We developed PCR-enzyme linked immunosorbent (ELISA) assays to detect Staphylococcus aureus enterotoxins A and B genes. The assays use internal biotin labelled oligonucleotides as capture probes for immobilizing and subsequently detecting target sequences on microtiter plates. The detection limits of the PCR ELISAs were approximately 250 gene copies, versus 2500 gene copies by agarose gel analysis. The sensitivity of the assays, as determined from a reference panel of 46 coded samples that included DNA purified from 31 different bacterial species and strains, SEA and SEB plasmid controls, and no-template controls was 100%. No cross-reactivity was observed with DNA from non-staphylococcal species. Using 27 clinical isolates of S. aureus, the SEA PCR-ELISA identified the enterotoxin A (sea) gene in 26 samples, and the SEB PCR-ELISA identified the enterotoxin B (seb) gene in all 27 samples. Compared with conventional antigen capture ELISAs for SEA and SEB toxins, the PCR-ELISAs showed overall superior detection limits. The sensitivity and specificity levels of the SEA PCR-ELISA and the SEA toxin ELISA were comparable within their respective detection thresholds, but the sensitivity and specificity of the SEB PCR-ELISA was much greater than that of SEB toxin ELISA. PMID- 10799268 TI - Subtyping of uncultured bartonellae using sequence comparison of 16 S/23 S rRNA intergenic spacer regions amplified directly from infected blood. AB - This study aimed to assess the usefulness of a PCR-based approach to the detection and differentiation of Bartonella strains in infected blood. The conservation of potential genus-specific PCR primer hybridisation sites within the 16 S/23 S rRNA gene intragenic spacer regions of Bartonella species was confirmed following sequence analysis of the intragenic spacer regions of four previously untested species. The extent of intra-species variation within the specific amplicons was assessed by comparison of sequences obtained from 17 strains of four Bartonella species. Eight sequence variants were obtained. Each species for which multiple strains were tested possessed at least two intragenic spacer regions variants, but the differences between these strains were markedly less than those observed inter-species. Sequence analysis was performed on 60 amplicons obtained from blood pellets collected from woodland rodent communities in which bartonella infections were known to be highly prevalent. Twelve variants were encountered, only five of which had been found among the studied isolates. Partial intragenic spacer region amplification followed by product sequence analysis offers a potentially sensitive and totally transferable means of inter- and intra-species differentiation of Bartonella strains, and its use in this study has broadened our knowledge of the genotypic spectrum of bartonellae associated with natural infections among UK woodland rodents. PMID- 10799270 TI - Multiplex PCR detection of Campylobacter jejuni and Arcobacter butzleri in food products. AB - Arcobacter is a recently described species, previously considered part of the Campylobacter family. A sensitive assay such as that provided by PCR could help to distinguish the closely related Arcobacter from Campylobacter. A PCR method to specifically detect both Campylobacter jejuni and Arcobacter butzleri in the same reaction tube has been developed. C. jejuni and A. butzleri were inoculated into a range of dairy products, raw and ready-to-eat foods. The presence of these two organisms was detected in these test foods by the multiplex PCR assay. A product of 159 bp was apparent when C. jejuni was present, while a 1223 bp product was seen when A. butzleri was present. When both organisms were present, both bands could be detected on the agarose gel. All organisms were confirmed by standard microbiological methods. There was complete agreement between the PCR and standard methods. This PCR assay will allow detection of both organisms within the same PCR tube and can be performed within an 8 h day. The presence of these two human pathogens, which are difficult to distinguish by standard biochemical methods, can now be identified using this PCR assay. PMID- 10799269 TI - Labelled trinucleotides as quantitative probes to identify Bacillus spp. using fluorescent in situ hybridization. AB - The number of nucleotide triplet repeats in 16 S rRNA sequences can be used for detection and identification of bacteria. Labelled TTT, GGG and ATA triplets were hybridized to the ribonucleic acid of Bacillus subtilis and Bacillus fusiformis whole-cells and the number of such triplets was quantified by synchronous fluorescence spectrometry. Each species was distinctly identified by specific ratios of labelled TTT, GGG and ATA triplets as well as characteristic fluorescence spectra. Notwithstanding the absence of intrinsic specificity, fluorescein-conjugated nucleotide triplet probes appear to be a useful tool for fluorescent spectrometric identification of micro-organisms through the quantitation of trinucleotide repeats. PMID- 10799271 TI - Improved diagnosis of porcine proliferative enteropathy caused by Lawsonia intracellularis using polymerase chain reaction-enzyme-linked oligosorbent assay (PCR-ELOSA). AB - Proliferative enteropathy (PE) caused by Lawsonia intracellularis is a major diarrheal disease affecting swine worldwide. Routine laboratory diagnosis of PE is done by amplification of L. intracellularis -specific DNA sequences by PCR followed by agarose gel electrophoresis and staining of PCR products with ethidium bromide. We report the development of an enzyme-linked oligosorbent assay (ELOSA) for specific identification of chromosomal L. intracellularis 328 bp PCR amplified products. The ELOSA involved determination of optical density value at 450 nm (OD(450)) after hybridization of biotin-labelled PCR products with an amine-modified internal oligonucleotide capture probe immobilized in microwell plates, and avidin-biotin-peroxidase complex. A positive ELOSA cut-off value of > or =0.375 was established using the mean OD(450)of negative control specimens plus three times the standard deviation. Using this value, the detection limit of PCR amplified L. intracellularis -specific products by ethidium bromide-stained agarose gel electrophoresis, Southern blot, and ELOSA were estimated to be 6.1 ng, between 0.8 and 3.0 ng, and 0.8 ng of DNA, respectively. Comparison of ethidium bromide-stained agarose gel analysis with ELOSA for detection of L. intracellularis -specific PCR products from 315 clinical specimens revealed 78% sensitivity, 100% specificity and 94% accuracy. The ELOSA produced a spectrophotometric signal that confirmed the authenticity of PCR products without subjective interpretation of ethidium bromide-stained PCR products after agarose gel electrophoresis. PMID- 10799272 TI - Detection of Yersinia pestis by pesticin fluorogenic probe-coupled PCR. AB - The <> assay (PE Applied Biosystems) combines PCR with concomitant release of fluorogenic nucleotides for immediate product detection by fluorometry. Yersinia pestis, the etiological agent of bubonic plague, expresses species-specific genes known to be located on two unique plasmids (9.6-kb pPCP and 100.9-kb pMT). Pesticin (pst) is a unique gene located on pPCP which encodes for a bacteriocin. Using fluorogenic probe coupled PCR as few as three copies of pst targets were detected from total Y. pestis genomic DNA. The pst probe used in this report was positive only for pesticinogenic isolates and did not show complementarity with Yersiniae nor with other bacteria targeted in this study suggesting, that the pst probe is very specific for Y. pestis. Under optimal conditions of Mg(2+)concentration and thermal cycle number, addition of extraneous DNA to respective assay mixtures had no effect on detection. PMID- 10799273 TI - Rare variants in the promoter of the fragile X syndrome gene (FMR1). AB - Fragile X syndrome, the most common form of familial mental retardation, is mainly caused by the expansion of an unstable region of CGG repeats in the 5' untranslated region of the FMR1 (Fragile X Mental Retardation-1) gene. Molecular tools to detect an abnormal CGG expansion in FMR1 include Southern blot hybridization and PCR amplification. Southern blotting with the StB12.3 probe and Eco RI/Eag I double digestion is widely used as a routine test for fragile X syndrome diagnosis in laboratories around the world. A patient with mental retardation of unknown origin showed absence of digestion for Eag I due to a 149C-->G substitution in the CpG island of the FMR1 gene, which destroys that restriction enzyme site. Screening for other changes around that region also detected a -154insGGC in a patient with a phenotype highly suggestive of fragile X syndrome but without CGG expansion. Expression studies did not show any abnormal changes in FMR1 function. In summary, we have identified two different changes (a C to G substitution at -149 and a GGC insertion at -154) in the promoter of the FMR1 gene. These are the first variants described in the promoter of the FMR1 gene. PMID- 10799274 TI - Quantitative polymerase chain (QPCR) reaction using the MIMIC approach to estimate Cryptosporidium parvum oocysts, an intestinal pathogen, in municipal water treatment sludge samples. AB - An accurate estimation of the number of Cryptosporidium parvum oocysts in water treatment plant sludge was determined using the Quantitative Polymerase Chain Reaction (QPCR) method. Approximately 8x10(6)purified viable oocysts were spiked into raw water and treated by conventional water treatment methods. The settled sludge was collected and the DNA extracted. The QPCR Mimic produced two competing products that were 300 and 435 base pairs in size. The log ratio of the products were used in the standard curve to determine a final estimation of oocysts in the sludge sample. The final number of oocysts in the sludge sample was estimated at 258 oocyst per two litres of treated water. This is the first time sludge from a water treatment process has been tested for presence of C. parvum oocysts, which is a known contaminant of drinking water. The QPCR method can be used to test other sludge samples and help estimate the sanitary risks associated with using sludge to fertilize agricultural lands. PMID- 10799276 TI - Uptake of metabolites by gonococci grown with lactate in a medium containing glucose: evidence for a surface location of the sialyltransferase. AB - Promotion of uptake of essential metabolites is a possible reason for the general stimulation of gonococcal metabolism which is caused by lactate (1 mM) in a defined medium containing glucose (5 mM). However, although uptake of(14)C adenine by gonococci [strain BS4(agar)] held for 4 or 7 min at 37 degrees C in Hanks balanced salt solution was increased for lactate treated gonococci compared with control organisms, uptake of(14)C glucose and(14)C proline under these conditions was unaffected. Hence, there is no evidence that lactate produces general stimulation of metabolite uptake. Unlike the other metabolites, cytidine 5'-monophospho-(14)CN-acetyl neuraminic acid (CMP-(14)CNANA), the substrate for sialylation of gonococcal lipopolysaccharide (LPS), was adsorbed in substantial quantities by gonococci held on ice for 6 min. Also, the increase in uptake of CMP-(14)CNANA at 37 degrees C over that adsorbed at 0 degrees C was much smaller (less than two-fold) than for the other compounds (4-30-fold). The substantial adsorption at 0 degrees C suggested a surface receptor for CMP-(14)CNANA. It is probably the sialyltransferase because a sialyltransferase deficient mutant, JB1, did not absorb CMP-(14)CNANA at 0 degrees C or take it up at 37 degrees C, in contrast to its parent strain, F62, which behaved similarly to strain BS4 (agar). This supports previous evidence for a surface location of the sialyltransferase. There was a small increase in adsorption of CMP-(14)CNANA in lactate treated gonococci indicating a slight increase in the surface enzyme. This could enhance LPS sialylation and hence affect pathogenicity. PMID- 10799275 TI - A reliability test of standard-based quantitative PCR: exogenous vs endogenous standards. AB - The quantitative measurement of gene expression requires consistent and reliable standards. At least two categories of standards, endogenous and exogenous, are currently used for quantitative PCR. The reliability of these two methods, however, has not been carefully compared. We hypothesized that a reliable quantitative PCR assay would be able to detect known dilutions of a given single stranded (ss-) cDNA. By measuring VEGF ss-cDNA copy numbers or signal ratios of GAPDH/VEGF in 10x and 100x diluted samples of two original ss-cDNA preparations, an exogenous recombinant DNA standard (a VEGF-mimic plasmid) and an endogenously expressed GAPDH standard were tested for their ability to detect dilution factors. Using the recombinant DNA standard, the dilution factor was detected as 10.3 and 135.0 in 10x and 100x diluted samples of the original CaSki cell ss cDNA, respectively. The detected dilution factors were 12.3 and 226.2, respectively, in 10x and 100x diluted ss-cDNA from U-251 MG cells. On the other hand, with the endogenous GAPDH standard, the dilution factors were detected as 2.7 and 8.0 in the same 10x and 100x dilutions of the original U-251 MG cell ss cDNA. Using the same endogenous GAPDH standard, the detected dilution factors were both 4.8 in 10x and 100x dilutions of the original CaSki cell ss-cDNA. It was also found that the number of endogenous copies of GAPDH mRNA was about 1000 times higher than VEGF. The high internal lockup ratio of GAPDH vs VEGF copy numbers and the requirement for additional primer pairs make the use of an abundant endogenous standard an unreliable choice in quantitative or semi quantitative PCR. In contrast, exogenous standard-based quantitative PCR was shown to be an accurate and reliable method for the quantitation of gene expression. PMID- 10799277 TI - An RNA helicase, RHIV -1, induced by porcine reproductive and respiratory syndrome virus (PRRSV) is mapped on porcine chromosome 10q13. AB - The impact of porcine reproductive and respiratory syndrome virus (PRRSV) infection on porcine alveolar macrophages (Mo) was examined by differential display reverse transcription PCR (DDRT-PCR). A PRRSV-induced expressed gene tag (EST) was used to isolate and identify a single cDNA clone from a library prepared from porcine peripheral blood. Rapid amplification of cDNA ends (RACE) was employed to clone a 1.5 kb fragment at the 5' end of the mRNA. DNA sequencing identified an open reading frame (ORF) of 2820 bp. Deduced amino acid sequence revealed the eight conserved domains characteristic of the DEAD/H box protein superfamily. The putative porcine RNA helicase induced by virus (RHIV -1) showed 84% amino acid similarity to human retinoic acid-induced gene (RIG-I). Porcine RHIV -1 transcripts were ubiquitously expressed in various pig tissues, while in PRRSV-infected pigs, higher expression was observed in several tissues persistent for PRRSV. These data indicate the association of PRRSV genome replication with enhaced host cell RNA helicase gene expression. Finally, the RHIV -1 gene was localized on porcine chromosome 10q13 between markers SSC25A02 and SWR334 via somatic cell panel and radiation hybrid (RH) mapping strategies. PMID- 10799278 TI - Isolation and characterization of a capsule-deficient mutant of Actinobacillus pleuropneumoniae serotype 1. AB - The capsular polysaccharides (CPS) play a major role in pathogenicity of Actinobacillus pleuroIpneumoniae, the causative agent of porcine pleuropneumonia. The purpose of the present study was to isolate a mutant in CPS biosynthesis by using a mini-Tn 10 transposon mutagenesis system and evaluate its adherence to host cells. One mutant apparently did not possess CPS as it did not react with a monoclonal antibody against A. pleuropneumoniae serotype 1 capsular antigen. Absence of capsule was confirmed by flow cytometry and also by transmission electron microscopy after polycationic ferritin labelling. The site of insertion of the mini-Tn 10 was determined and found to be in the cpxC gene. Its gene product, CpxC, is a protein involved in polysaccharide transport across the cytoplasmic membrane during CPS biosynthesis. Use of piglet tracheal frozen sections indicated that the CPS mutant adhered significantly (P=0.0001) more than the parent strain. The non-capsular mutant was less virulent in pigs compared to the parent strain and showed no mortality in experimentally infected pigs. The CPS mutant was however resistant to pig serum. This CPS mutant is the first A. pleuropneumoniae mutant in a CPS transport gene. It is also the first time that adherence of a CPS mutant of A. pleuropneumoniae is evaluated. Our observations indicate that capsular polysaccharides of A. pleuropneumoniae serotype 1 are not involved in adherence to piglet tracheal frozen sections but rather mask, at least in part, the adhesive functions. PMID- 10799279 TI - Presence of the LEE (locus of enterocyte effacement) in pig attaching and effacing Escherichia coli and characterization of eae, espA, espB and espD genes of PEPEC (pig EPEC) strain 1390. AB - In the present study, attaching and effacing Escherichia coli (AEEC) O45 isolates from post-weaning pigs with diarrhoea were examined for the presence of the LEE (locus of enterocyte effacement) using various DNA probes derived from the LEE of human enteropathogenic E. coli (EPEC) strain E2348/69. The LEE fragment was conserved among the eae -positive pig isolates. The attaching and effacing activity of PEPEC (pig EPEC) O45 isolates is highly correlated with the presence of the LEE. Nevertheless, for some PEPEC isolates, the insertion site of the LEE is different or has diverged during evolution. The presence of the LEE fragment in PEPEC isolates provides further evidence that the LEE region is conserved among AEEC of different animal origins. In addition, the nucleotide sequence of the region containing the eae gene and esp genes of a pig AEEC isolate, strain 1390, was determined. Among examined Eae proteins, Eae of strain 1390 showed the highest similarity with Eae belonging to the beta intimin group such as the Eae of rabbit AEEC. Moreover, all pig strains that produced attaching and effacing lesions in piglets and pig ileal explants belonged to the beta intimin group. The deduced amino acid sequences of the EspA, EspB and EspD proteins of strain 1390 showed particularly strong homology to those of AEEC strains presenting a beta intimin allele. Thus, pig AEEC possess the LEE sequences, and for the strain 1390, sequences of the eae and esp regions are related to those of other AEEC, in particular, strains presenting a beta intimin allele, such as the rabbit AEEC. PMID- 10799280 TI - Characterization of a DNA region containing 5'-(CAAT)(n)-3' DNA sequences involved in lipooligosaccharide biosynthesis in Haemophilus somnus. AB - Repetitive tetranucleotide sequences of 5'-(CAAT)(n)-3' have been identified at the 5' end of an open reading frame (ORF) named lob1 from Haemophilus somnus strain 738. Based on sequence analysis, lob1 has 59% DNA homology to lex2B, which is involved in lipooligosaccharide (LOS) biosynthesis in H. influenzae. We now report that the number of 5'-CAAT-3' repeats in lob1 varied from 31-35, but that 94% of colonies contained 33 repeats of 5'-CAAT-3' downstream of two potential start codons, as determined by DNA sequence analysis of the 5'-CAAT-3' region from individual colonies. If transcription began with the start codon closest to the 5'-CAAT-3' repeats, a protein of 34.5 kDa would be encoded when 33 repeats were present. However, we could not establish a correlation between the number of 5'-CAAT-3' repeats in lob1 with a specific LOS electrophoretic profile or reactivity with two LOS monoclonal antibodies, indicating multiple genes control LOS phase variation in H. somnus. Complementation of strain 129Pt with lob1 containing 33 5 '-CAAT-3' repeats in shuttle vector pLS88 resulted in transformants 129Pt(pLSlob1-33A) and 129Pt(pLSlob1-33B), both of which demonstrated the same altered LOS electrophoretic profile. Unlike strain 129Pt, both transformants underwent limited LOS phase variation, which correlated with variation in the number of 5'-CAAT-3' repeats in pLSlob1-33. Nanoelectrospray mass spectrometry of O-deacylated LOS indicated that transformant 129Pt(pLSlob1 33A) LOS was composed of a different distribution of glycoforms than LOS of the parent strain. The ratio of glucose to galactose changed from 1:2 in strain 129Pt LOS to 2:1 in transformant 129Pt(pLSlob1-33A) LOS, as determined by gas chromatography-mass spectrometry. Nuclear magnetic resonance spectroscopy confirmed and extended these observations. Transformant 129Pt(pLSlob1-33A) was constitutively more reactive in colony immunoblotting to polyclonal antiserum made to purified strain 738 LOS, and was more susceptible to complement-mediated killing in the presence of anti-738 LOS serum than parent strain 129Pt. Based on these results, Lob1 appears to be a phase variable galactosyl transferase involved in LOS biosynthesis in H. somnus. PMID- 10799281 TI - Secretion of cytokines by human macrophages upon infection by pathogenic and non pathogenic mycobacteria. AB - The interaction of various pathogenic (Mycobacterium tuberculosis, M. avium, M. kansasii, M. xenopi), and non-pathogenic mycobacteria (M. smegmatis, M. phlei) with human macrophages at the level of macrophage cytokine expression (TNFalpha, IL1, IL6 and GM-CSF) was investigated. Both for TNFalpha and GM-CSF, the lowest levels were obtained with pathogenic mycobacterial species, whereas about 2-8 times higher levels were observed for non-pathogenic species. Contrary to the above, the differences for IL6 and IL1 were not marked, although IL6 appeared to be more elevated for non-pathogenic species. Heat-killed bacteria induced a lower level of the cytokines for all the three cytokines assayed (TNFalpha, IL6 and IL1), except for M. tuberculosis for whom a significantly higher proportion of TNFalpha was induced by killed bacilli. The RT-PCR experiments performed on M. avium (as a low inducer of the cytokines) and M. smegmatis (as a high inducer of the cytokines) showed that the differences observed among pathogenic vs non pathogenic strains were also reflected at the transcriptional level for TNFalpha and to a lesser extent for IL6, but not for IL1. This investigation underlined important differences existing between the pathogenic and non-pathogenic species, particularly as regards TNFalpha and GM-CSF. PMID- 10799282 TI - Effects of tetrandrine on growth factor-induced DNA synthesis and proliferative response of rat pulmonary artery smooth muscle cells. AB - The objective of the present study was to investigate the effect of tetrandrine (a plant alkaloid isolated from Stephenia tetrandra) on growth factor-induced DNA synthesis and proliferative responses of rat pulmonary artery smooth muscle cells. Male rat and bovine pulmonary artery smooth muscle cells (PASMC) were cultured in Medium 199 containing FBS (10%). DNA synthesis was monitored from [(3)H]-thymidine uptake and cell proliferation by direct cell counting. In the present study FBS (1% v/v) caused a small increase in DNA synthesis above basal levels in rat and bovine PASMC (6% and 11% respectively). Platelet-derived growth factor (PDGF, 50 ng/ml), fibroblast growth factor (FGF, 50 ng/ml) or interleukin 1alpha (IL-1alpha, 100 pg/ml) alone increased rat PASMC proliferation (69-85%) and DNA synthesis above basal levels (76-92%). The addition of these growth factors in combination with FBS (1%) resulted in higher increases in DNA synthesis above basal levels (rat PASMC:PDGF, 465%; FGF, 421%; IL-1alpha, 406%; bovine PASMC:PDGF, 279%). Tetrandrine (10(-5) M) inhibited FBS (10%)-induced rat PASMC proliferation (90.5%) and DNA synthesis (89.0%). Tetrandrine significantly inhibited cell proliferation (86.5-98.5%) and DNA synthesis (79.9-89.0%) induced by FBS (1%) in combination with one of the following mitogens; PDGF (50 ng/ml), FGF (50 ng/ml), IL-1alpha (100 pg/ml). The inhibitory effects of tetrandrine were observed between 10(-6) and 10(-5)M and PASMC viability was not affected by tetrandrine below 3x10(-5) m. In summary, these results suggest that tetrandrine can exert anti-proliferative effects against a range of mitogenic stimuli for vascular smooth muscle cells in vitro. Such effects may contribute to the inhibitory effect of tetrandrine on pulmonary vascular remodelling associated with pulmonary hypertension. PMID- 10799283 TI - Lipopolysaccharide-induced lung injury in mice. I. Concomitant evaluation of inflammatory cells and haemorrhagic lung damage. AB - Intratracheal instillation of lipopolysaccharide (LPS) induces an inflammatory response characterized by infiltration of polymorphonuclear neutrophils (PMNs) into the extracellular matrix and by the release of mediators that play a fundamental role in lung damage. In the present study, we developed a mouse model which allows correlation of the inflammatory response and haemorrhagic tissue injury in the same animal. In particular, the different steps of the inflammatory response and tissue damage were evaluated by the analysis of three parameters: myeloperoxidase (MPO) activity in the parenchyma, reflecting PMNs accumulation into the lung, inflammatory cells count in the bronchoalveolar lavage fluid (BALF), reflecting their extravasation, and total haemoglobin estimation in BALF, a marker of haemorrhagic tissue damage consequent to PMNs degranulation. In our experimental conditions, intra-tracheal administration of 10 microg/mouse of LPS evoked an increase of MPO activity in the lung at 4 h (131%) and 6 h (147%) from endotoxin challenge. A significant increase of PMNs in the BALF was noticed at these times with a plateau between the 12nd and 24th h. PMN accumulation produced a time-dependent haemorrhagic lung damage until 24 h after LPS injection (4 h: +38%; 6 h: +23%; 12 h: +44%; 24 h: +129% increase of haemoglobin concentration in the BALF vs. control). Lung injury was also assessed histopathologically. Twenty four hours after the challenge, diffuse alveolar haemorrhage, as well as PMN recruitment in the interstitium and alveolus were observed in the LPS group. This model was pharmacologically characterized by pretreatment of LPS-treated mice with antiinflammatory drugs acting on different steps of the <>. We demonstrated that: a) betamethasone (1, 3, 10, 30 mg/kg p.o.) reduced in a dose-dependent manner the MPO activity, the number of inflammatory cells and, at the same time, lung injury; b) pentoxifylline, a TNFalpha production inhibitor (200 mg/kg i.p.), inhibited PMN extravasation and lung haemorrhage but it was not able to reduce MPO activity in the lung; c) L-680,833, an anti-elastase compound (30 mg/kg po), decreased significantly only the haemorrhagic lung damage; d) indomethacin, a non steroidal antiinflammatory drug (5 mg/kg p.o.), did not show any effect on any of the parameters considered. In conclusion, our in vivo mouse model is a practical alternative to animal models of ARDS (Adult Respiratory Distress Syndrome) recently described and it permits to dissect and to characterize the different steps of PMNs infiltration and, at the same time, the damage caused by their activation. PMID- 10799284 TI - Lipopolysaccharide-induced lung injury in mice. II. Evaluation of functional damage in isolated parenchyma strips. AB - Pulmonary inflammatory diseases are characterized by changes in airway responsiveness. This phenomenon is commonly related to the action of inflammatory mediators produced by infiltrated leukocytes. The aim of this study was to investigate in an ex vivo experimental model the effect of acute instillation of lipopolysaccharide (bacterial endotoxin; LPS) on lung parenchyma contractility. We firstly characterized the responsiveness of isolated murine lung to airway stimuli. Murine parenchymal strips were found to be mainly sensitive to 5 hydroxytryptamine (5-HT) while the cholinergic agonist, methacholine (MCh), evoked a smaller contractile response. 5-HT responsiveness was inhibited by methysergide. No significant parenchymal contraction was evoked by histamine, substance P and bradykinin. Lung responsiveness to 5-HT was significantly reduced by in vivo LPS treatment and this effect was only partially paralleled by leukocyte infiltration. In addition, LPS-induced hyporesponsiveness was significantly inhibited by betamethasone (BMS) or pentoxifylline (PTX) pretreatment suggesting that 5-HT lung hyporesponsiveness could be mediated by LPS-induced inflammatory mediators such as inflammatory cytokines. PMID- 10799285 TI - Effect of Ca(2+)-independent mechanisms on the hypoxic relaxation of guinea-pig tracheal rings. AB - Hypoxia induces bronchodilation in vivo and in vitro, but the mechanisms are still unclear. To evaluate whether an extra- or intracellular free Ca(2+) ion is involved in the mechanisms of hypoxic relaxation, we simultaneously measured cytosolic Ca(2+)levels and tensions in both intact and denuded guinea-pig tracheal strips precontracted with histamine (100 microM), and assessed the effect of hypoxia on guinea-pig tracheal rings precontracted with okadaic acid (10 microM) and calyculin-A (0.1 approximately 10 microM) under an extracellular Ca(2+)-free state. The exposure of tracheal rings to hypoxia induced an immediate decrease of tracheal tension without decrease in intracellular free Ca(2+)levels. In the presence of okadaic acid but not calyculin-A, hypoxic air exposure caused significant transient reductions in tracheal tone. Further, thapsigargin (5 microM or 10 microM) did not affect hypoxic bronchodilation, suggesting that the release of intracellular Ca(2+) does not take a role in hypoxic bronchodilation. Hypoxic dilation decreased ATP content in epithelium-intact rings but not epithelium-denuded rings, indicating a relationship between hypoxic dilation and change of adenine nucleotide in epithelium-intact rings. Our findings indicate that the epithelium dependent mechanisms of hypoxic relaxation of guinea pig tracheal rings preconstricted with histamine may not be related to the mobilization of extra and intra-cellular Ca(2+). PMID- 10799286 TI - Attenuation of pulmonary vascular hypertension and cardiac hypertrophy with sitaxsentan sodium, an orally active ET(A) receptor antagonist. AB - Effects of sitaxsentan (TBC11251), an orally active, highly selective antagonist of endothelin A receptors, were examined on the development and maintenance of pulmonary hypertension, pulmonary vascular remodeling, and cardiac hypertrophy in the rat. The pulmonary vasoconstrictor response to acute hypoxia (10% O(2)for 90 min) was prevented with sitaxsentan (5 mg/kg infused iv 10 min prior to the onset of hypoxia) while BQ-788 (a specific endothelin B receptor antagonist) was without effect. The same dose of sitaxsentan delivered iv 50 min after the onset of hypoxia reversed the established pulmonary vasoconstriction. In a 2-week model of hypoxia using 10% O(2), treatment with sitaxsentan (15 mg/kg per day in drinking water) attenuated pulmonary hypertension and the associated right ventricular hypertrophy, and prevented the remodeling of small pulmonary arteries (50-100 microM) without affecting systemic arterial blood pressure or heart rate. Institution of sitaxsentan treatment (15 and 30 mg/kg per day in drinking water) for 4 weeks after 2 weeks of untreated hypoxia produced a significant, dose dependent reversal of the established pulmonary hypertension, right heart hypertrophy, and pulmonary vascular remodeling despite continued hypoxic exposure. Sitaxsentan blocked increased plasma endothelin levels in the prevention protocol but did not affect the established elevated levels in the intervention study. Sitaxsentan dose dependently (10 and 50 mg/kg per day in the drinking water) attenuated right ventricular systolic pressure, right heart hypertrophy, and pulmonary vascular remodeling observed 3 weeks after a single subcutaneous injection of monocrotaline. These findings support the hypothesis that endothelin-1 plays a significant role in the development of pulmonary hypertension, pulmonary vascular remodeling, and the associated cardiac hypertrophy, and further suggest that specific endothelin-A receptor blockade may be useful in the treatment of pulmonary hypertension of diverse etiologies. PMID- 10799287 TI - Structure of O-linked GlcNAc transferase: mediator of glycan-dependent signaling. PMID- 10799288 TI - An O-acetylated sialyl-Tn is involved in ovarian cancer-associated antigenicity. AB - We reported previously that a monoclonal antibody, 6G9, raised against bovine submaxillary mucin (BSM) reacted with mucinous ovarian cancer and recognized tumor-associated sialylated carbohydrate antigens. To obtain structural information on the carbohydrate antigens recognized by 6G9, the reactivity of several mucins and carbohydrates with the antibody was determined by ELISA. Exoglycosidase digestion of BSM showed that 6G9 recognized Sia as a nonreducing monosaccharide but neither Gal nor GlcNAc. Reactivity of BSM with 6G9 decreased markedly on de-O-acetylation of BSM in mild alkali, and O-acetyl Sia obtained from BSM reacted with the antibody, indicating the presence of O-acetyl groups on Sia in the epitope. A sialyl-Tn structure located in the epitope was also demonstrated by the findings that de-O-acetylated BSM retained weak but significant reactivity with 6G9 and that ovine submaxillary mucin, major sugar chains of which are sialyl-Tn, reacted with 6G9 stronger than de-O-acetylated BSM. Furthermore, weak reactivity of NeuAcalpha2 --> 6GalNAc prepared from BSM demonstrated that 6G9 recognized the sialyl-Tn structure, but the modification of Sia with O-acetyl groups was essential for the recognition. The failure of 9-O acetyl NeuAc, synthesized chemically, to react with the antibody implied that 6G9 recognized sialyl-Tn with O-acetylation on Sia distinct from C-9 O-acetylation. PMID- 10799289 TI - Glycosaminoglycan-binding properties and secondary structure of the C-terminus of netrin-1. AB - Netrins are soluble neurite-outgrowth-promoting proteins related to the laminin B2 chain. Since these proteins and their receptor DCC (the "deleted in colorectal carcinoma" gene product) bind heparin, glycosaminoglycans may modulate their biological actions in a similar fashion as described for several other ligand receptor systems. Here we show that a polypeptide encompassing the C-terminal cluster of basic amino acids of netrin-1 (i) adopts an alpha-helical conformation in water-trifluoroethanol mixtures according to circular dichroism experiments and (ii) binds electrostatically to heparin with high affinity under physiological ionic conditions (K(D) = 15 nM for the binding to immobilized heparin according to surface plasmon resonance, K(D) = 50 nM in solution as determined with isothermal titration calorimetry). These data indicate that the cluster of basic amino acids at the C-terminus of netrin-1 forms an alpha-helical structural element which can contribute to the glycosaminoglycan-binding activity of this neurotrophic guidance molecule. PMID- 10799290 TI - Stable interdomain interaction within the cytoplasmic domain of CD45 increases enzyme stability. AB - CD45 is a leukocyte-specific, two domain transmembrane tyrosine phosphatase. Co purification of a recombinant protein containing the first phosphatase domain of CD45 (6His-D1) with a recombinant protein containing the second phosphatase domain (GST-D2) from E. coli indicated a stable interaction which resulted in increased stability of the active phosphatase domain present in 6His-D1. This interaction was not dependent on the acidic region unique to CD45 domain 2, but was affected by a destabilizing point mutation (Q1180G) in GST-D2. CD45 domain 2 enhanced phosphatase activity of the first domain in the full length cytoplasmic domain protein, whereas a chimeric protein with the SH2 domain of p56(lck) in place of the CD45 C-terminal region did not. Thus the C-terminal domain of CD45 associates with the N-terminal domain and this stabilizes the active phosphatase domain. A single destabilizing point mutation in the second domain is sufficient to attenuate this effect. PMID- 10799291 TI - Phosphorylation of Cdc20/fizzy negatively regulates the mammalian cyclosome/APC in the mitotic checkpoint. AB - The cyclosome/anaphase promoting complex (APC) is a multisubunit ubiquitin ligase that targets mitotic regulators for degradation in exit from mitosis. It is activated at the end of mitosis by phosphorylation and association with the WD-40 protein Cdc20/Fizzy and is then kept active in the G1 phase by association with Cdh1/Hct1. The mitotic checkpoint system that keeps cells with defective spindles from leaving mitosis interacts with Cdc20 and prevents its stimulatory action on the cyclosome. The activity of Cdh1 is negatively regulated by phosphorylation, while the abundance of Cdc20 is cell cycle regulated, with a peak in M-phase. Cdc20 is also phosphorylated in G2/M and in mitotically arrested cells, but the role of phosphorylation remained unknown. Here we show that phosphorylation of Cdc20 by Cdk1/cyclin B abrogates its ability to activate cyclosome/APC from mitotic HeLa cells. A nonphosphorylatable derivative of Cdc20 stimulates cyclin ubiquitin ligation in extracts from nocodazole-arrested cells to a much greater extent than does wild-type Cdc20. It is suggested that inhibitory phosphorylation of Cdc20/Fizzy may have a role in keeping the cyclosome inactive in early mitosis and under conditions of mitotic checkpoint arrest. PMID- 10799292 TI - PCCX1, a novel DNA-binding protein with PHD finger and CXXC domain, is regulated by proteolysis. AB - We identified a novel gene PCCX1 that encoded a nuclear protein carrying a PHD finger, a CXXC domain, and an acidic region. The CXXC domain was found to be sufficient for binding to DNA. The acidic region exhibited a high transactivation ability, but the full-length protein was inactive due to regions which inhibited the acidic region, including the C-terminal region. We examined the expression of PCCX1 during cellular aging and immortalization of SV40-transformed human fibroblasts. PCCX1 mRNA was expressed constitutively through stages of cellular aging and immortalization, but at the protein level, a shorter form lacking the C terminal region appeared as the cells approached crisis. These results suggested that PCCX1 was activated by proteolytic cleavage, which removed the C-terminal inhibitory region. PMID- 10799293 TI - Differential involvement of vacuolar H(+)-ATPase in the refilling of thapsigargin and agonist-mobilized Ca(2+) stores. AB - Our objective was to evaluate the role of vacuolar H(+)-ATPase and proton gradients in the refilling of Ca(2+) stores in fura-2-loaded pancreatic acinar cells. Once depleted with a high level of ACh, the Ca(2+) stores were replenished with a Ca(2+)-containing solution. The degree of refilling was estimated with a second release in response to either ACh (ACh-releasable store) or thapsigargin (thapsigargin-releasable store), a specific inhibitor of the endoplasmic reticulum Ca(2+) pumps. Both the protonophore nigericin and folimycin, a specific inhibitor of the vacuolar H(+)-ATPase, reduced reuptake into the ACh-mobilized stores but not into the thapsigargin-releasable pools. These treatments effectively dissipated the subcellular pH gradients (revealed by confocal observation of the distribution of a marker for acidic compartments), and did not impair the [Ca(2+)](i) response to ACh in control cells. Our results indicate that thapsigargin and ACh release heterogeneous Ca(2+) stores which are differently operated by vacuolar proton ATPase. PMID- 10799294 TI - Suppression of endothelin-converting enzyme-1 during buccal mucosal ulcer healing: effect of chronic alcohol ingestion. AB - Among the factors affecting the efficiency of soft oral tissue healing is endothelin-1 (ET-1), a potent vasoactive peptide produced from a biologically inactive big ET-1 by the action of endothelin-converting enzyme-1 (ECE-1). We investigated the expression of ECE-1 during buccal mucosal ulcer healing in rats maintained for 5 weeks on alcohol containing or control diet. The mucosal activity of ECE-1, characterized by sensitivity to phosphoramidon, was associated with microsomal fraction and showed an elevated (3.1-fold) level in the alcohol diet group. Moreover, the ulcer onset in the alcohol group was reflected in a 39% greater expression of ECE-1 activity, and was accompanied by a 1.4-fold greater increase in TNF-alpha and a 2.5-fold greater enhancement in epithelial cell apoptosis. While in both groups the ulcer healing was associated with a decrease in buccal mucosal expression of ECE-1, as well as a decline in TNF-alpha and apoptosis, the changes were significantly slower in the alcohol diet group and manifested by a 40% delay in healing. Thus, chronic alcohol ingestion leads to up regulation of ECE-1 expression, induction of TNF-alpha, and triggering apoptotic events that delay the mucosal repair. PMID- 10799295 TI - Exposure to extremely low frequency magnetic fields suppresses x-ray-induced transformation in mouse C3H10T1/2 cells. AB - We designed and manufactured equipment for exposure of cultured cells to extremely low frequency magnetic fields (ELFMF) at 5, 50, and 400 mT and examined the effect of ELFMF on cellular transformation in mouse C3H10T1/2 cells (clone 8). Transformed foci, Type II and Type III, were independently counted as transformants. The cells were exposed to ELFMF alone at 5, 50, and 400 mT for 24 h or X-irradiated with 3 Gy followed by the ELFMF exposure. No significant difference in the transformation was observed between sham-exposed control and the ELFMF exposure from 5 to 400 mT. The transformation frequency for X-rays plus ELFMF was decreasing compared with X-rays alone. When 12-O-tetra-decanoylphorbol 13-acetate (TPA) was contained in the medium throughout the experiment, the transformation frequency by X-rays alone was elevated more. In the combined treatment with X-rays followed by ELFMF, the transformation frequency was slightly decreased at 50 and 400 mT even in the medium containing TPA. The long term exposure at 5 mT suppressed both spontaneous and X-ray-induced transformations significantly. It is well known that overexpressing protein kinase C (PKC) failed to yield identifiable transformation of foci induced by ionizing radiation. We demonstrated previously that exposure to high-density ELFMF induced expression of several genes through an increase in PKC activity. From these results, it is suggested that ELFMF might suppress X-ray-induced transformation through activation of PKC by ELFMF. PMID- 10799296 TI - Ras GTPase is essential for fas-mediated activation of phospholipase D in A20 cells. AB - We have previously reported that Fas cross-linking resulted in an increase in phospholipase D activity in A20 murine cells (J.-S. Han et al., Arch. Biochem. Biophys. 367, 233-239, 1999). In an attempt to explore the Fas downstream factor contributing to the activation of phospholipase D, we have investigated the possible involvement of a small GTP biding protein Ras in signaling events that were triggered by Fas cross-linking. Upon adenoviral expression of dominant negative mutant of Ras (N17Ras), an increase in phospholipase D activity by anti Fas monoclonal antibody was diminished. Also, the Fas downstream signaling events triggered by Fas cross-linking such as the activation of phosphatidylcholine specific phospholipase C, the increase in diacylglycerol level, and the translocation of protein kinase C to membrane fraction were all reduced by N17Ras expression. When parallel experiments were performed with manumycin-A, a Ras farnensyltransferase inhibitor, almost identical inhibitory effects on Fas downstream signaling were exhibited. These data suggest that Ras GTPase is essential in transmitting phospholipase D activation signal induced by Fas cross linking and is located at phosphatidylcholine-specific phospholipase C upstream in Fas signaling cascades. PMID- 10799297 TI - Cloning and expression of beta,beta-carotene 15,15'-dioxygenase. AB - beta,beta-Carotene 15,15'-dioxygenase cleaves beta-carotene into two molecules of retinal and is therefore the key enzyme in beta-carotene metabolism to vitamin A. In the present study, it was possible to enrich the chicken beta,beta-carotene 15,15'-dioxygenase to such an extent that partial amino acid sequence information could be obtained to design degenerate oligonucleotides. With RT-PCR a cDNA fragment could be obtained and used subsequently in a radioactive screening of a chicken duodenal expression library. We cloned the first eukaryotic beta,beta carotene 15,15'-dioxygenase which symmetrically cleaves beta-carotene at the 15,15'-double bond. PMID- 10799298 TI - Susceptibility of the prion protein to enzymic phosphorylation. AB - Ten protein kinases have been assayed for their ability to phosphorylate in vitro the recombinant bovine PrP (25-242) (rbPrP). Substantial phosphorylation was observed with PKC, CK2, and two tyrosine kinases, Lyn and c-Fgr. With regard to CK2, phosphorylation occurs at Ser 154 with a stoichiometry of about 0.1 mol phosphate/mol rbPrP, which is doubled by mild heat treatment of rbPrP. Heat also reduces the overall protein ellipticity, suggesting that reversibly unfolded conformers are more susceptible to phosphorylation. Our data disclose the possibility that phosphorylation might modulate PrP biological activity. PMID- 10799299 TI - PI3-K/AKT regulation of NF-kappaB signaling events in suppression of TNF-induced apoptosis. AB - We found that in MCF-7 breast carcinoma cells, PI3K and Akt suppressed a dose dependent induction of apoptosis by tumor necrosis factor alpha (TNF). PI3K and Akt stimulated NF-kappaB activation in a dose-dependent manner, suggesting a common link between these two pathways. TNF has been shown to activate both an apoptotic cascade, as well as a cell survival signal through NF-kappaB. PI3K and AKT cell survival signaling were correlated with increased TNF-stimulated NF kappaB activity in MCF-7 cells. We demonstrate that while both TNFR1 and NIK are partially involved in Akt-induced NF-kappaB stimulation, a dominant negative IkappaBalpha completely blocked Akt-NF-kappaB cross-talk. PI3K-Akt signaling activated NF-kappaB through both TNFR signaling-dependent and -independent mechanisms, potentially representing a mechanism by which Akt functions to suppress apoptosis in cancer. PMID- 10799300 TI - The ovine CCAAT-enhancer binding protein delta gene: cloning, characterization, and species-specific autoregulation. AB - Transcription factors belonging to the CCAAT-enhancer binding protein (C/EBP) family have been implicated in the regulation of gene expression during growth, differentiation, apoptosis, and inflammation. Autoregulation is relatively common in the modulation of C/EBP gene expression and, for the human and murine C/EBPalpha, it is known that species-specific autoregulatory mechanisms operate. It is therefore essential to investigate the autoregulation of additional C/EBP genes from a wider range of different species to gauge the degree of commonality, or otherwise, which exists. As an important step towards this goal, we report here the cloning and the characterisation of the ovine C/EBPdelta gene (ovC/EBPdelta) and analysis of its promoter region. Transient transfection assays reveal that ovC/EBPdelta acts as a transcriptional activator. Although several motifs that are characteristic of C/EBPdelta genes are conserved in the ovine sequence, including the basic region, leucine zipper, and activation domains, two regions have been identified that are specifically absent in the ovine and bovine homologues. The ovC/EBPdelta promoter is active in both the hepatoma Hep3B and the mammary epithelial HC11 cell lines, induced by the cytokine interleukin-6 and autoregulated by mechanisms that are potentially different from those described for the rat promoter. These results suggest that, in common with C/EBPalpha, the C/EBPdelta genes may also be subject to autoregulation by distinct species specific mechanisms. PMID- 10799301 TI - Nitric oxide inhibits inducible nitric oxide synthase mRNA expression in RAW 264.7 macrophages. AB - Using cultured murine RAW 264.7 macrophages, the present study investigates the influence of nitric oxide (NO) on the expression of the inducible NO synthase (iNOS) enzyme at the transcriptional level. Incubation of cells with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) led to a marked increase in iNOS mRNA levels. Inhibition of LPS/IFN-gamma-induced NO synthesis with the L-arginine analogue N(G)-monomethyl-L-arginine (L-NMMA) was accompanied by a significant up-regulation of iNOS mRNA that was reversed in the presence of the NO donor sodium nitroprusside (SNP). Treatment of cells with SNP alone decreased LPS/IFN-gamma-induced iNOS mRNA levels in a concentration-dependent manner. The inhibitory effect of SNP on iNOS mRNA expression was not prevented by 1H-[1,2, 4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ), a selective inhibitor of the soluble guanylyl cyclase. In agreement with this finding, incubation of cells with the membrane-permeable cyclic GMP analogue 8-bromo cyclic GMP left LPS/IFN gamma-induced iNOS mRNA expression virtually unaltered. Together, our results demonstrate that both iNOS-derived and exogenous NO exert an inhibitory effect on the expression of iNOS by a mechanism independent of the soluble guanylyl cyclase/cyclic GMP pathway. In conclusion, NO may control the extent of iNOS mRNA expression by a negative autoregulatory feedback. PMID- 10799302 TI - Mutation analysis of cadherin-4 reveals amino acid residues of EC1 important for the structure and function. AB - To clarify the structural basis of the cell adhesion activity of cadherins, we examined the effects of point mutations of well-conserved amino acid residues in the extracellular domain 1 of cadherin-4 (Cdh4) on the adhesion properties by alanine scanning mutagenesis. Mutations of two well-conserved aromatic amino acid residues in the extracellular domain 1 resulted in abnormal processing of Cdh4 molecules and no cell adhesion activity, whereas mutations of the corresponding aromatic amino acids in the extracellular domain 2 did not show these effects, suggesting a role for the two residues in the extracellular domain 1 in the folding and/or intracellular transport processes of Cdh4. Mutations of the amino acid residues suspected to be involved in strand dimer formation resulted in loss or significant decrease in cell adhesion activity. The mutant Cdh4s showed weak concentration at cell-cell adhesion sites and chemical cross-linking suggested that the strand dimer formation was actually impaired in the mutants. These results are consistent with the zipper model, in which the extracellular domain 1 of Cdh4 has intrinsic strand dimer formation activity in addition to adhesion dimer formation activity, both of which are involved in cell adhesion activity. The zipper model, however, needs further improvement to fully account for the present results. PMID- 10799303 TI - Dexamethasone inhibits tumor necrosis factor-alpha-induced expression of macrophage inflammatory protein-2 and adhesion of neutrophils to endothelial cells. AB - Macrophage inflammatory protein-2 (MIP-2) belongs to the C-X-C subfamily of chemokines and appears to play an important role in cytokine-induced inflammatory and immune cell-mediated responses. We found that tumor necrosis factor-alpha (TNF-alpha) time- and dose-dependently increased gene and protein expression of MIP-2 in endothelial cells. Moreover, it was observed that dexamethasone treatment inhibited endothelial cell expression of MIP-2 in response to TNF-alpha stimulation and markedly reduced the number of adherent neutrophils. Moreover, we found that a monoclonal antibody against murine MIP-2 abolished neutrophil adhesion to TNF-alpha-activated endothelial cells. These data demonstrate that TNF-alpha induces expression of MIP-2 in endothelial cells and support the hypothesis that the anti-inflammatory action of dexamethasone may, at least in part, be attributable to an inhibition of MIP-2 induction on cytokine-activated endothelial cells. PMID- 10799304 TI - Hyperosmotic NaCl and urea synergistically regulate the expression of the UT-A2 urea transporter in vitro and in vivo. AB - The UT-A2 urea transporter is involved in the recycling of urea through the kidney, a process required to maintain high osmotic gradients. Dehydration increases UT-A2 expression in vivo. The tissue distribution of UT-A2 suggested that hyperosmolarity, and not vasopressin, might mediate this effect. We have analyzed the regulation of UT-A2 expression by ambiant osmolarity both in vitro (mIMCD3 cell line) and in vivo (rat kidney medulla). The UT-A2 mRNA was found to be synergistically up-regulated by a combination of NaCl and urea. Curiously, the UT-A2 protein was undetectable in this hypertonic culture condition, or after transfection of the UT-A2 cDNA, whereas it could be detected in HEK-293 transfected cells. Treating rats with furosemide, a diuretic which decreases the kidney interstitium osmolarity without affecting vasopressin levels, led to decreased levels of the UT-A2 protein. Our results show that the UT-A2 urea transporter is regulated by hyperosmolarity both in vitro and in vivo. PMID- 10799305 TI - Natriuretic peptide receptor-A negatively regulates mitogen-activated protein kinase and proliferation of mesangial cells: role of cGMP-dependent protein kinase. AB - We have examined the effect of atrial natriuretic peptide (ANP) and its guanylyl cyclase/natriuretic peptide receptor-A (NPRA) on mitogen-activated protein kinase/extracellular signal-regulated kinase 2 (MAPK/ERK2) activity in rat mesangial cells overexpressing NPRA. Agonist hormones such as platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), angiotensin II (ANG II), and endothelin-1 (ET-1) stimulated 2.5- to 3.5-fold immunoreactive MAPK/ERK2 activity in these cells. ANP inhibited agonist-stimulated activity of MAPK/ERK2 by 65-75% in cells overexpressing NPRA, whereas in vector-transfected cells, its inhibitory effect was only 18-20%. NPRA antagonist A71915 and KT5823, a specific inhibitor of cGMP-dependent protein kinase (PKG) completely reversed the inhibitory effect of ANP on MAPK/ERK2 activity. ANP also inhibited the PDGF stimulated [(3)H]thymidine uptake by almost 70% in cells overexpressing NPRA, as compared with only 20-25% inhibition in vector-transfected cells. These results demonstrate that ANP/NPRA system negatively regulates MAPK/ERK2 activity and proliferation of mesangial cells in a PKG-dependent manner. PMID- 10799306 TI - The nuclear-encoded SDH2-RPS14 precursor is proteolytically processed between SDH2 and RPS14 to generate maize mitochondrial RPS14. AB - In maize, the functional gene encoding mitochondrial ribosomal protein S14 (rps14) has been translocated to the nucleus where it became integrated between both exons of a gene encoding the iron-sulfur subunit of succinate dehydrogenase (sdh2). Two transcripts are generated from this locus by alternative splicing. One transcript encodes a precursor for a functional SDH2 protein, while the second transcript encodes a chimeric SDH2(t)-RPS14 precursor protein. In this paper we show that the same mitochondrial targeting presequence is able to direct the import of both precursors into isolated mitochondria and is removed during import. This processing event generates a 28 kDa protein from the SDH2 precursor, which corresponds to the iron-sulfur subunit of respiratory complex II present in maize mitochondria. In addition to cleavage of the presequence, the chimeric precursor undergoes proteolytical processing between SDH2 and RPS14. This processing generates RPS14, which is found assembled into mitochondrial ribosomes, and a truncated SDH2 protein which is degraded. Therefore, our results support a role of the SDH2 domain in the chimeric precursor only in providing a mitochondrial targeting function for RPS14. PMID- 10799307 TI - Enhanced macrophage uptake of elastase-modified high-density lipoproteins. AB - Incubation of human HDL (d = 1.063-1.21 g/ml) with monocyte-derived elastase causes selective proteolysis of apoA-II and apoA-I apolipoproteins. We have found that elastase-digested HDL (ED-HDL) bind to J774-A1 murine macrophages with enhanced affinity and are internalized and degraded at a rate threefold higher than that of native HDL. Unlike oxidized LDL and HDL and proteolytically modified LDL, the uptake of ED-HDL lipoproteins does not affect the cellular lipid biosynthesis nor modify the cell lipid content. The cell surface binding of (125)I-ED-HDL can be competed by native HDL but not by acetylated LDL, consistent with the idea that ED-HDL are recognized by the class B type I scavenger receptor. The liberation of elastase by lipid-engorging macrophages is regarded as an important event during atherogenesis. By enhancing the cellular uptake of HDL this process can lead to a local decrease of antiatherogenic HDL particles. PMID- 10799308 TI - Overexpression of human UDP-glucose pyrophosphorylase rescues galactose-1 phosphate uridyltransferase-deficient yeast. AB - To better understand the pathophysiology of galactose-1-phosphate uridyltransferase (GALT) deficiency in humans, we studied the mechanisms by which a GALT-deficient yeast survived on galactose medium. Under normal conditions, GALT-deficient yeast cannot grow in medium that contains 0.2% galactose as the sole carbohydrate, a phenotype of Gal(-). We isolated revertants from a GALT deficient yeast by direct selection for growth in galactose, a phenotype of Gal(+). Comparison of gene expression profiles among wild-type and revertant strains on galactose medium revealed that the revertant down-regulated genes encoding enzymes including galactokinase, galactose permease, and UDP-galactose-4 epimerase (the GAL regulon). By contrast, the revertant strain up-regulated the gene for UDP-glucose pyrophosphorylase, UGP1. There was reduced accumulation of galactose-1-phosphate in the galactose-grown revertant cells when compared to the GALT-deficient parent cells. In vitro biochemical analysis showed that UDP glucose pyrophosphorylase had bifunctional properties and could catalyze the conversion of galactose-1-phosphate to UDP-galactose in the presence of UTP. To test if augmented expression of this gene could produce a Gal(+) phenotype in the GALT-deficient parent cells, we overexpressed the yeast UGP1 and the human homolog, hUGP2 in the mutant strain. The Gal(-) yeast transformed with either UGP1 or hUGP2 regained their ability to grow on galactose. We conclude that revertant can grow on galactose medium by reducing the accumulation of toxic precursors through down-regulation of the GAL regulon and up-regulation of the UGP1 gene. We speculate that increased expression of hUGP2 in humans could alleviate poor outcomes in humans with classic galactosemia. PMID- 10799309 TI - Structure of the cytoplasmic domain of p23 in solution: implications for the formation of COPI vesicles. AB - Coatomer, the coat protein complex of coat protein (COPI) vesicles, is involved in the budding of these vesicles. Its interaction with the cytoplasmic domains of some p24-family members, type I transmembrane proteins of the Golgi, has been shown to induce a conformational change of coatomer that initiates polymerization of the complex. From stoichiometrical data it is likely that interaction of coatomer with the small tail domains involves an oligomeric form of the p24 proteins. Here we present the structure of peptide analogs of the cytoplasmic domain of p23, a member of the p24 family, as determined by two-dimensional nuclear magnetic resonance spectroscopy in the presence of 2,2,2 trifluoroethanol. An improved strategy for structure calculation revealed that the tail domain peptides form alpha-helices and adopt a tetrameric state. Based on these results we propose an initial model for the binding of coatomer by p23 and the induced conformational change of coatomer that results in its polymerization, curvature of the Golgi membrane to form a bud, and finally a COPI coated vesicle. PMID- 10799310 TI - Enhanced expression of Silencer of death domains (SODD/BAG-4) in pancreatic cancer. AB - Pancreatic cancers are resistant to TNF-alpha-mediated apoptosis. Silencer of death domains (SODD) binds to TNF-alpha receptor TNFR-1, and prevents spontaneous self-association of death domains and inappropriate receptor signaling. In addition, overexpression of SODD suppresses TNF-alpha-induced cell death. In this report, we demonstrate increased SODD mRNA levels in pancreatic cancer (n = 30) in comparison to normal control tissues (n = 20, P < 0.01). In situ hybridization analysis revealed SODD expression predominantly in the cancer cells within the pancreatic tumor mass. In contrast, SODD mRNA levels were not different (P > 0.05) in four other gastrointestinal cancers (liver, esophagus, stomach, colon) compared with the corresponding normal tissues. These findings indicate that in contrast to other gastrointestinal malignancies, in pancreatic cancer SODD might block TNF-alpha-mediated apoptosis which may influence the growth of pancreatic cancer cells. PMID- 10799311 TI - Neuregulin induces the rapid association of focal adhesion kinase with the erbB2 erbB3 receptor complex in schwann cells. AB - Neuregulins signal cells by binding to an activating hetero- and homodimeric forms of the neuregulin receptors HER2 (erbB2), HER3 (erbB3), and HER4 (erbB4). Axonally derived neuregulin signals myelin forming cells of the central and peripheral nervous systems through different receptor complexes: oligodendrocytes through erbB2/erbB4 heterodimers and Schwann cells through erbB2/erbB3 heterodimers. Since the leading edge of myelinating cells interacts directly with the axonal surface, we were interested in determining if signaling molecules localized at the leading edge associate with activated neuregulin receptors. We found a novel association between neuregulin receptors and focal adhesion kinase (FAK) in primary cultures of Schwann cells. Following stimulation with ligand, maximal binding of FAK to HER2 occurred by 1 min whereas maximal binding to HER3 was delayed to approximately 7 min. FAK is localized in focal adhesions of Schwann cells. We have previously shown HER2 and HER3 are distributed evenly throughout the plasmalemma. Neuregulins thus use FAK to transmit intracellular signals and the differential kinetics of FAK association with individual neuregulin receptors, as well as its restricted subcellular localization, may play a role in specifying biologic responses. PMID- 10799313 TI - Sustained enhancement of Ca(2+) influx by glibenclamide induces apoptosis in RINm5F cells. AB - Cytosolic Ca(2+) elevations are known to be involved in triggering apoptosis in many tissues, but the effect of sustained enhancement of Ca(2+) influx on apoptosis in beta cells remains unknown. We have found that the viability of RINm5F cells is decreased dose-dependently by continuous exposure to glibenclamide at concentrations from 10(-7) to 10(-4) M, and that this effect is partially ameliorated by pretreatment with cycloheximide. Electrophoresis of the cells exposed to glibenclamide revealed ladder-like fragmentation characteristic of apoptosis, and which also is suppressed by cycloheximide pretreatment. By using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining, we detected increased DNA fragmentation in the nuclei of the cells exposed to glibenclamide, and staining with Hoechst 33342 and propidium iodide showed a dose-dependent increase in the number of cells with the chromatin condensation and fragmentation in their nuclei that is characteristic of apoptosis. The effects of glibenclamide on cell viability and apoptotic cell death were partially inhibited by treatment with Ca(2+) channel blocker, and by reducing the extracellular Ca(2+) concentration during glibenclamide exposure, suggesting that they may be derived from increased Ca(2+) influx. Furthermore, only the percentage of apoptotic cells, and not that of necrotic cells, increased with the increasing intracellular Ca(2+) concentration during glibenclamide exposure. In conclusion, we have demonstrated that the sustained enhancement of Ca(2+) influx caused by glibenclamide exposure can induce apoptotic cell death in a pure beta cell line. PMID- 10799312 TI - Neuronal differentiation of cryopreserved neural progenitor cells derived from mouse embryonic stem cells. AB - Embryonic stem cells (ES cells) are developmentally pluripotent cells isolated from pre-implantation mammalian embryos. In cell culture ES cells can be easily differentiated to generate cultures of neural progenitors. We present a simple method for the cryopreservation of these ES-derived neural progenitors. Cryopreserved neural progenitor stocks can be thawed, expanded with FGF2, and differentiated into functional neurons. This method will facilitate studies using ES-derived neural progenitor cells as a cell culture model system for neural development and differentiation. It will also aid studies designed to test the ability of these progenitor cells to functionally engraft and repair damaged neural tissue. PMID- 10799314 TI - Osmomechanical regulation of membrane trafficking in polarized cells. AB - The regulation of membrane trafficking is thought to be predominantly under the control of agonist-receptor transduction pathways. In the present study, osmomechanical stress due to swelling, a condition often accompanying cell activation, was shown to induce multiple membrane trafficking pathways in polarized absorptive epithelial cells in the absence of agonists. Osmomechanical stress activated rapidly (seconds) pathways of calcium-dependent membrane insertion into the basolateral domain, pathways of calcium-independent membrane retrieval from the basolateral domain, and a novel pathway of transcytosis (transcellular) between basolateral and apical cell domains. These pathways appear to underlie the transfer and regulation of transport proteins amongst cell compartments. This broad affect of osmomechanical stress on trafficking pathways may reflect a global mechanism for redistribution of transport proteins and other membrane components amongst cell compartments during states of mechanical stress. PMID- 10799315 TI - Recombinant expression of a selective blocker of M(1) muscarinic receptors. AB - Mamba venoms contain peptides with high selectivity for muscarinic receptors. Due to the limited availability of the M(1) muscarinic receptor-selective MT7 or m1 toxin 1, the peptide was expressed in Sf9 cells using a synthetic cDNA and purified. The isolated peptide had over four orders of magnitude higher affinity for the M(1) compared to M(2)-M(5) muscarinic receptors. The peptide strongly inhibited Ca(2+) mobilisation through recombinant and endogenously expressed M(1) receptors, having no effect on the function of the other subtypes. The MT7 peptide provides a unique tool for identification and functional characterisation of M(1) receptors in cells and tissues. PMID- 10799316 TI - Transient expression of megakaryocyte-derived protein immunoreactive with an antiserum to cartilage oligomeric matrix protein in developing rat liver. AB - We analyzed a megakaryocyte-derived protein immunoreactive with an antiserum to cartilage oligomeric matrix protein (COMP) in the developing rat liver. Staining with the anti-COMP antiserum in the developing rat liver increased during embryogenesis, and was strongest in the livers of 17-day-old embryos. However, staining in the liver was not detected at eight days after birth or thereafter. The stained cells were found to be megakaryocytes. We partially purified the protein showing cross-reaction with the antiserum to COMP from a megakaryocyte rich cells fraction in 17-day-old embrionic rat livers. The molecular weight of this protein (approximately 95 kDa) was close to the molecular weight of COMP (105 kDa). Amplification of an RT-PCR fragment (225 bp) corresponding to part of COMP mRNA was detected in cartilage, but not in megakaryocytes of fetal liver or bone marrow. Based on these results, the fetal rat liver megakaryocyte-derived protein that reacted with the antiserum against COMP was thought to contain a common epitope with COMP from cartilage, but to be a different protein from COMP. PMID- 10799317 TI - Fenofibrate and rosiglitazone lower serum triglycerides with opposing effects on body weight. AB - Activators of peroxisome proliferator activated receptors (PPARs) are effective drugs to improve the metabolic abnormalities linking hypertriglyceridemia to diabetes, hyperglycemia, insulin-resistance, and atherosclerosis. We compared the pharmacological profile of a PPARalpha activator, fenofibrate, and a PPARgamma activator, rosiglitazone, on serum parameters, target gene expression, and body weight gain in (fa/fa) fatty Zucker rats and db/db mice as well as their association in db/db mice. Fenofibrate faithfully modified the expression of PPARalpha responsive genes. Rosiglitazone increased adipose tissue aP2 mRNA in both models while increasing liver acyl CoA oxidase mRNA in db/db mice but not in fatty Zucker rats. Both drugs lowered serum triglycerides yet rosiglitazone markedly increased body weight gain while fenofibrate decreased body weight gain in fatty Zucker rats. KRP 297, which has been reported to be a PPARalpha and gamma co-activator, also affected serum triglycerides and insulin in fatty Zucker rats although no change in body weight gain was noted. These results serve to clearly differentiate the metabolic finality of two distinct classes of drugs, as well as their corresponding nuclear receptors, having similar effects on serum triglycerides. PMID- 10799318 TI - Analysis of hABC1 gene 5' end: additional peptide sequence, promoter region, and four polymorphisms. AB - Evidence linking mutations in ATP-binding-cassette transporter gene 1 (ABC1) to Tangier disease suggests it functions in the active transport of free cholesterol out of cells. Since its mRNA level is regulated in response to cellular cholesterol stores it is of interest to explore its promoter response elements, and to investigate polymorphisms for their contributions to the prevalence of low levels of HDL in the population that promotes premature coronary heart disease. Investigation of the 5' end of the gene by 5' RACE analysis revealed 455 nucleotides additional to published sequences, and predicts another 60 amino acid N-terminal residues, resulting in a 2261-residue protein. Protein sequence analysis predicts a membrane-spanning region and possible signal peptide. The 5' flanking region was located by a Human Research Project BLAST search. This region contains regulatory elements that potentially control ABC1 gene expression. In addition to numerous SP1 binding sites there are four putative sterol regulatory elements (SREs). Our studies uncovered three single nucleotide substitution polymorphisms, one in the promoter region and two in the 5' untranslated region (5'UTR), plus an insertion/deletion polymorphism. PMID- 10799319 TI - Characterization of hPRP4 kinase activation: potential role in signaling. AB - Mitogen-activated protein kinases (MAPKs) and cyclin-dependent kinases (CDKs) are important proline-directed Ser/Thr kinases that play distinct roles in cell differentiation and proliferation. hPRP4 (pre-mRNA processing gene), a human homologue of S. pombe Prp4, is a recently isolated CDK-like kinase with homology to MAPKs. Little is known about the mRNA processing function of hPRP4 or about the signaling pathways with which it is associated. hPRP4 is expressed in a variety of human tissues with the highest expression in the brain, lung and liver. In this paper, we characterize the activation of hPRP4 in COS-7 cells and show that hPRP4 also possesses a transcription factor activation function. hPRP4 is activated by epidermal growth factor (EGF) or forskolin treatment, but not tetradecanoyl phorbol acetate (TPA) nor ultraviolet (UV) irradiation. Activated hPRP4 phosphorylates residue Thr-417 on Elk-1 resulting in Elk-1 activation. This site of Elk-1 phosphorylation is distinct from that of other MAPKs. Coexpression of hPRP4 with an Elk-1 reporter construct causes trans activation of the reporter. These findings suggest that hPRP4, a CDK-like kinase related to MAPKs, may play a distinct role in signal transduction in addition to its role in mRNA processing. PMID- 10799320 TI - Purification and polymerization properties of two lethal yeast actin mutants. AB - The budding yeast Saccharomyces cerevisiae contains a single actin gene and the gene product, actin, is essential for growth. Two mutants of yeast actin that do not support yeast growth were prepared from yeast by coexpressing the mutant and a 6-histidine-tagged wild-type actin followed by separation of the wild-type and mutant actin using Ni-NTA chromatography as described elsewhere [Buzan, J., Du, J., Karpova, T., and Frieden, C. (1999) Proc. Natl. Acad. Sci. USA 96, 2823 2827]. The mutations, in muscle actin numbering, were at positions 334 (Glu334Lys) and 168 (Gly168Arg) and were chosen based on phenotypic changes observed in the behavior of actin mutants of Caenorhabditis elegans. Glu334 is located on the surface of actin between subdomains 1 and 3 while Gly168 is located in a region near actin-actin contacts in the actin filament. The Glu334Lys mutant polymerized slightly faster than wild-type yeast actin, suggesting that loss of interactions with some actin binding protein, rather than loss of actin-actin contacts, was responsible for its inability to support yeast growth. The Gly168Arg mutant polymerized at a rate similar to wild-type but the extent was considerably less, kinetic characteristics suggesting a high critical concentration (ca. 4 microM) without a large change in the ability to form nuclei for the nucleation-elongation process. PMID- 10799321 TI - The N-terminal domains of cyclin-dependent kinase inhibitory proteins block the phosphorylation of cdk2/Cyclin E by the CDK-activating kinase. AB - It has been suggested that binding of p27 and p21 kinase inhibitory proteins (KIPs) to cyclin-dependent kinases (cdks) render them inaccessible to cdk activating kinase (CAK), presumably by steric hindrance by the C-terminal residues. However, this common mechanism of inhibition is inconsistent with the known structural divergence in the p27 and p21 C-terminal domains. Therefore, we studied the direct binding of N-terminal minimal domain of p27 (amino acids 28 81) to cdk2/cyclin E. An unlabeled p27 minimal domain, mutated in the N-terminal LFG motif, was unable to compete with a labeled minimal domain for binding to cdk2/cyclin E. The p27 and its minimal domain inhibited CAK-mediated phosphorylation of cdk2/cyclin E. This inhibitory effect was significantly diminished with p27 minimal domain mutated in the LFG motif. A synthetic peptide, ACRRLFGPVDSE, from the N-terminal residues 17-28 of p21, was also a potent inhibitor of CAK-mediated cdk2/cyclin E phosphorylation. Taken together, these results show that anchoring of p27 or p21 KIPs to cyclin E via the N-terminal LFG containing motif can block CAK access to its cdk2/cyclin E substrate. PMID- 10799322 TI - Cloning, characterization, and expression of the human TIN-ag-RP gene encoding a novel putative extracellular matrix protein. AB - The human gene encoding a novel tubulointerstitial nephritis antigen (TIN-ag) related protein (TIN-ag-RP) was isolated, and its genomic organization was determined. BLAST searches revealed the highest degree of homology to several mammalian TIN-ag orthologues, and a weak homology to cathepsin B-like proteases. The 12 kb gene was mapped by fluorescence in situ hybridization to chromosome 1p34.2-3, a locus neither related to that of the human TIN-ag (6p11.2-12) nor to that of cathepsin B (8p22-23.1). The TIN-ag-RP is encoded in ten exons with introns ranging from 83 bp to 4 kb. In addition, the gene contained one exon in the 5'UTR, but none in the 3'UTR. Five of the 10 splice sites of the TIN-ag-RP gene were fully conserved when compared to a related gene of C. elegans, whereas only one splice site was identical to those found in cathepsin B genes. Furthermore, human TIN-ag-RP tagged with the T7-epitope, was expressed in HeLa cells, and was found to be localized in vesicular compartments as well as secreted into the medium suggesting the involvement of the endosomal trafficking pathway. Based on the high degree of homology of the amino acid sequences and genomic organization between TIN-ag-RP and TIN-ag, we suggest that both molecules may form a distinct group or family of TIN-ag-like proteins. PMID- 10799323 TI - Differential regulation of the carbonic anhydrase II gene expression by hormonal nuclear receptors in monocytic cells: identification of the retinoic acid response element. AB - The Carbonic Anhydrase II (CAII) gene that encodes an enzyme involved in proton production is expressed in several cell types including monocyte/macrophage derived osteoclasts. We have analyzed the regulation of the chicken CAII promoter/reporter construct by nuclear hormone receptors of the VDR subfamily in HD11 avian macrophages. The CAII expression is stimulated by 1, 25 dihydroxyvitamin D(3) but not by 9-cis retinoic acid and repressed by VDR overexpression due to RXR squelching. It is also stimulated by all-trans retinoic acid only when RARalpha is overexpressed, and is dependent on a RARE located in the distal part of the promoter and bound by RARalpha homodimer. Finally, in macrophages, unlike in erythrocytes, the CAII promoter is unresponsive to thyroid hormone. Our results demonstrate the first retinoic acid response element in the CAII promoter and show that according to cell type, different nuclear receptors of the VDR subfamily can regulate the CAII gene. PMID- 10799324 TI - Binding of human mitochondrial transcription factor A, an HMG box protein, to a four-way DNA junction. AB - Mitochondrial transcription factor A (mtTFA), the only known transcription factor in mitochondria, is also implicated in maintenance of mitochondrial genome although little is elucidated about its molecular basis. mtTFA is a member of HMG box proteins family. Some HMG proteins bind with high affinity to four-way DNA junctions that mimic a Holliday structure, a putative intermediate in DNA recombination. To explore possible involvement of a Holliday-like structure in the maintenance of mitochondrial genome, we examine the binding of recombinant human mtTFA to a synthetic four-way DNA junction. The human mtTFA binds to the four-way DNA junction with an approximately 10-fold higher affinity than to the corresponding linear duplex DNA and with essentially the same affinity as to a 40 mer DNA containing the human mitochondrial light strand promoter sequence. The mtTFA binds to the four-way as a monomer. Both of the two HMG box domains of human mtTFA are required for the high affinity binding to the four-way junction. PMID- 10799325 TI - Endothelial proliferation, migration, and differentiation are blunted by conditionally expressed protein kinase C pseudosubstrate peptides. AB - Peptides based on the pseudosubstrate (PS) sequence of conventional protein kinase C isoenzymes (alpha, beta, gamma) specifically inhibit PKC activity in permeabilized cells, but whether PS can be used to study the role of PKC in the proliferation or migration of intact endothelial cells (EC) and angiogenesis is unknown. Peptides based on the PKCeta pseudosubstrate (etaPS) sequence were 3.5- to 8-fold more potent in inhibiting the PKCalpha, delta, epsilon, or eta kinase activity than was the peptide based on the PKCalpha pseudosubstrate (alphaPS) sequence. Thus, etaPS was conditionally overexpressed in intact EC and compared to alphaPS. Serum-induced growth of EC expressing etaPS was significantly slower than that of control EC. etaPS EC demonstrated slower rate of serum stimulated migration than that of either control or alphaPS EC. Expression of either etaPS or alphaPS produced slower rates of PMA induced EC migration, as compared to control EC. In an in vitro three-dimensional assay in which EC organize into capillary tubules, the EC that expressed etaPS formed fewer such tubules. This study shows that pseudosubstrate inhibitors derived from PKCeta are more potent both in vitro and in vivo than one based on the conventional isoenzyme PKCalpha. These data further support a role for PKC in proliferation and migration of intact EC, and angiogenesis. PMID- 10799326 TI - Inhibitory effect of nitric oxide on voltage-dependent calcium currents in rat dorsal root ganglion cells. AB - The effect of nitric oxide (NO) on calcium current (I(Ca)) and intracellular calcium concentration ([Ca(2+)](i)) in primarily cultured dorsal root ganglion (DRG) neurons was investigated from neonatal rats. I(Ca) and [Ca(2+)](i) were simultaneously recorded using perforated-patch technique in combination with fluorescence measurement from single DRG neurons. NO donors, sodium nitroprusside (SNP) and S-nitro-N-acetylpenicillamine (SNAP), inhibited I(Ca) in small-diameter neurons without significant change in voltage-dependence of activation and activation time constants. SNP and SNAP also reduced the transient [Ca(2+)](i) peak accompanied by I(Ca). Inhibition by NO was reproducible, but gradually desensitized. In some DRG neurons, SNP and SNAP increased basal [Ca(2+)](i) in concentration of 10 microM with little effect on NO-induced inhibition of I(Ca). 8-Br-cGMP, a permeable cGMP analog, mimicked the effects of SNP and SNAP. These results suggest that, in DRG neurons, NO has inhibitory effect on I(Ca), which is independent of NO-induced increase of basal [Ca(2+)](i), through cGMP-dependent pathway. PMID- 10799327 TI - Identification of six CAG repeat domains into the human chromosomic region 12q24.1. AB - Six different domains of CAG repeats from a human chromosome 12 specific cosmid library were identified, cloned, and sequenced. These CAG repeat domains were localized into the human chromosomic region 12q24.1. Five of them constitute repeat candidates for expansions in autosomal dominant neurological disorders with genetic anticipation, and they can also contribute to the chromosome walking in the human genome project. PMID- 10799328 TI - ATP hydrolysis by a CFTR domain: pharmacology and effects of G551D mutation. AB - Residues 417-830 of the cystic fibrosis transmembrane conductance regulator (CFTR) were expressed as a glutathione-S-transferase fusion protein. This fusion protein, NBD1/R/GST, contains the regulatory and first nucleotide binding domains of CFTR. NBD1/R/GST hydrolyzed ATP with a K(M) (60 microM) and V(max) (330 nmol/min/mg) that differed from those reported for CFTR and for a peptide containing CFTR residues 433-589. The ATPase inhibitor profile of NBD1/R/GST indicates that CFTR resembles P-glycoprotein with respect to the NBD1 ATPase catalytic mechanism. ATP hydrolysis by NBD1/R/GST was unaffected by genistein, glybenclamide, and other agents known to affect CFTR's chloride channel function, suggesting that these agents do not act by directly influencing the ATPase function of NBD1. The disease-causing mutation, G551D, reduced ATP hydrolysis by NBD1/R/GST by increasing the K(M) for ATP fourfold. This suggests that when G551D occurs in patients with cystic fibrosis, it affects CFTR function by reducing the affinity of NBD1 for ATP. PMID- 10799329 TI - Mouse myosin X: molecular architecture and tissue expression as revealed by northern blot and in situ hybridization analyses. AB - The structure of the coding region of mouse myosin X cDNA was determined. The predicted protein sequence indicated an approximately 240 kDa molecular mass with 2062 amino acids. When aligned with the structure predicted for calf myosin X (GenBank Accession No. U55042), extremely highly conserved pleckstrin homology domains and a myosin tail homology 4 domain were apparent in the tail region, suggesting their importance for myosin X's function. Northern blot analysis revealed the existence of a myosin X mRNA, 8.7 kb in size, in various mouse tissues, while a similar size of human type myosin X mRNA was recognized mainly in the testis. In addition to the adult-type transcripts in mice, a smaller embryo-specific mRNA, 4.8 kb in size, was identified in early to late embryonic stages, suggesting the presence of a shorter myosin X isoform in mouse embryos. In situ hybridization experiments with mouse testis revealed that myosin X mRNA was restricted to Sertoli cells at stages VIII-X of the spermatogenesis cycle, suggesting that myosin X is implicated in the supporting cells during the spermatid morphogenesis. PMID- 10799330 TI - Vectors with hidden cloning sites. AB - A general strategy is described for using the cleavage site of restriction enzymes in vectors for cloning regardless of how many sites the given enzymes have in the vector. The application of this method allows one to open any vector at its cloning site with protruding ends which can be compatible with almost every commercially available Class II restriction enzyme. By employing this method, the laborious construction of new vectors can be simplified considerably. This general strategy is based on the known ability of Class IIS restriction enzymes to cut any sequence located outside of their recognition site; the introduction of a linker containing recognition site(s) for Class IIS restriction enzyme(s), not present originally in the vector, gives rise to the possibility of opening the vector so as to produce overhangs of arbitrary sequence. In particular, when a symmetrical short sequence representing the protruding end of any Class II enzyme is situated at the cutting position of the Class IIS enzyme, cleavage with the Class IIS enzyme exposes the hitherto hidden, "unique" cloning site. This technique is demonstrated by cloning the cDNA of the multidrug resistance protein to an expression vector. PMID- 10799331 TI - Elevated levels of RanBP7 mRNA in colorectal carcinoma are associated with increased proliferation and are similar to the transcription pattern of the proto oncogene c-myc. AB - We have used suppression subtractive hybridisation, "in silico" cloning and reverse Northern dot blot analysis to identify significant up-regulation of RanBP7 transcription in a human colorectal carcinoma. Quantitative RT-PCR analyses using the Taqman system demonstrated that RanBP7 mRNA levels were elevated in 47/75 colorectal tumours. There was no significant difference in 17 matched normal and tumour pairs and reduced levels in 11. Since RanBP7 specifies a key member of nuclear transport receptors responsible for the nuclear import of histone H1 and ribosomal proteins, we investigated whether this up-regulation might be proliferation-associated. RanBP7 mRNA copy numbers were significantly correlated with those of proliferating cell nuclear antigen in both normal and cancer tissue. Interestingly, the transcription pattern of the proto-oncogene c myc showed a similar correlation with PCNA mRNA. Our results highlight the need for the careful interpretation of quantitative data that compare mRNA levels in normal and cancer tissue. PMID- 10799332 TI - Adenovirus-mediated transfer of caspase-8 in combination with superrepressor of NF-kappaB drastically induced apoptosis in gliomas. AB - Inhibition of NF-kappaB in the presence of tumor necrosis factor-alpha (TNF) is supposed to be a promising cancer therapeutic approach, since it disrupts the protective mechanism of NF-kappaB activated by TNF. To test this approach in gliomas, we introduced a superrepressor of NF-kappaB, an N-terminal deleted form of inhibitor kappa B alpha (IkappaBdN) gene, to human glioma cells (U251 and U 373MG) via adenoviral vector (Adv) in the presence of TNF. U-373MG cells were refractory to TNF-induced apoptosis even when they were transduced with the IkappaBdN gene. On the other hand, transduction of IkappaBdN drastically augmented caspase-8-mediated apoptosis in U-373MG cells. Similar results were obtained in U251 cells. Cotransduction of IkappaBdN and caspase-8 induced cleavage of PARP. Taken together, Adv-mediated transfer of IkappaBdN plus caspase 8 may be a promising therapeutic approach to treat gliomas. PMID- 10799333 TI - Kinetic determination of focal adhesion protein formation. AB - I examined the binding kinetics between integrin (alpha(IIb)beta(3)) and purified focal adhesion proteins, including alpha-actinin, filamin, vinculin, talin, and F actin. Using static light-scatter technique, I observed affinities of the order talin > filamin > F-actin > alpha-actinin > (talin when bound to vinculin) which were lower when integrin was complexed with fibronectin. No binding between integrin and vinculin was detected. The calculated dissociation constants (K(d)) ranged between 0.4 microM and 5 microM. These results in part confirm previously published data using different methods. The modest affinity with which the focal adhesion proteins interact in vitro might be indicative of how cells, e.g., thrombocytes, gain a high degree of versatility and velocity. PMID- 10799334 TI - Evidence that E-cadherin may be a target for cadmium toxicity in epithelial cells. AB - E-cadherin is a Ca(2+)-dependent cell adhesion molecule that plays an important role in the development and maintenance of epithelial polarity and barrier function. This commentary describes the results of recent studies showing that the environmental pollutant Cd(2+) can damage the E-cadherin-dependent junctions between many types of epithelial cells and reviews the evidence indicating that this effect results from the direct interaction of Cd(2+) with the E-cadherin molecule. In addition, the implications of these findings with respect to the mechanisms of Cd(2+) toxicity in specific target organs such as lung, kidney, bone, and the vascular endothelium are discussed. PMID- 10799335 TI - The spontaneously hypertensive rat as a model of human cardiovascular disease: evidence of exacerbated cardiopulmonary injury and oxidative stress from inhaled emission particulate matter. AB - Cardiovascular disease is considered a probable risk factor of particulate matter (PM)-related mortality and morbidity. It was hypothesized that rats with hereditary systemic hypertension and underlying cardiac disease would be more susceptible than healthy normotensive rats to pulmonary injury from inhaled residual oil fly ash (ROFA) PM. Eight spontaneously hypertensive (SH) and eight normotensive Wistar-Kyoto (WKY) rats (12-13 weeks old) were implanted with radiotelemetry transmitters on Day -10 for measurement of electrocardiographic (ECG) waveforms. These and other nonimplanted rats were exposed to filtered air or ROFA (containing leachable toxic levels of metals) on Day 0 by nose-only inhalation (ROFA, 15 mg/m(3) x 6 h/day x 3 days). ECGs were monitored during both exposure and nonexposure periods. At 0 or 18 h post-ROFA exposure, rats were assessed for airway hyperreactivity, pulmonary and cardiac histological lesions, bronchoalveolar lavage fluid (BALF) markers of lung injury, oxidative stress, and cytokine gene expression. Comparisons were made in two areas: (1) underlying cardiopulmonary complications of control SH rats in comparison to control WKY rats; and (2) ROFA-induced cardiopulmonary injury/inflammation and oxidative burden. With respect to the first area, control air-exposed SH rats had higher lung and left ventricular weights when compared to age-matched WKY rats. SH rats had hyporeactive airways to acetylcholine challenge. Lung histology revealed the presence of activated macrophages, neutrophils, and hemorrhage in control SHrats. Consistently, levels of BALF protein, macrophages, neutrophils, and red blood cells were also higher in SH rats. Thiobarbituric acid-reactive material in the BALF of air-exposed SH rats was significantly higher than that of WKY rats. Lung inflammation and lesions were mirrored in the higher basal levels of pulmonary cytokine mRNA expression. Cardiomyopathy and monocytic cell infiltration were apparent in the left ventricle of SH rats, along with increased cytokine expression. ECG demonstrated a depressed ST segment area in SH rats. With regard to the second area of comparison (ROFA-exposed rats), pulmonary histology indicated a slightly exacerbated pulmonary lesions including inflammatory response to ROFA in SH rats compared to WKY rats and ROFA-induced increases in BALF protein and albumin were significantly higher in SH rats than in WKY rats. In addition, ROFA caused an increase in BALF red blood cells in SH rats, indicating increased hemorrhage in the alveolar parenchyma. The number of alveolar macrophages increased more dramatically in SH rats following ROFA exposure, whereas neutrophils increased similarly in both strains. Despite greater pulmonary injury in SH rats, ROFA-induced increases in BALF GSH, ascorbate, and uric acid were attenuated when compared to WKY rats. ROFA inhalation exposure was associated with similar increases in pulmonary mRNA expression of IL-6, cellular fibronectin, and glucose-6-phosphate dehydrogenase (relative to that of beta-actin) in both rat strains. The expression of MIP-2 was increased in WKY but attenuated in SH rats. Thus, SH rats have underlying cardiac and pulmonary complications. When exposed to ROFA, SH rats exhibited exacerbated pulmonary injury, an attenuated antioxidant response, and acute depression in ST segment area of ECG, which is consistent with a greater susceptibility to adverse health effects of fugitive combustion PM. This study shows that the SH rat is a potentially useful model of genetically determined susceptibility with pulmonary and cardiovascular complications. PMID- 10799336 TI - Uncoupling between bone formation and resorption in ovariectomized rats with chronic cadmium exposure. AB - Osteoporosis, osteomalacia, and pathological fractures are characteristic features of Itai-Itai disease. The mechanisms of bone damage caused by cadmium (Cd) exposure have not been fully clarified. We investigated skeletal changes in ovariectomized rats with chronic Cd exposure, using bone histomorphometry and mechanical tests. Female Sprague-Dawley rats at the age of 8 weeks were ovariectomized. Eight weeks after ovariectomy, the rats were divided into two groups: Cd-OVX group (n = 15), ovariectomized rats given cadmium chloride (CdCl(2), 0.18 mg/rat) ip three times a week for 28 weeks; Cont-OVX group (n = 10), ovariectomized rats given distilled water alone for 28 weeks. Cd-OVX rats had a significant increase in serum concentration of intact osteocalcine and showed numerical but not significant increase in urinary excretion of deoxypyridinoline despite a significant decrease in glomerular filtration rate to 40% of the value in Cont-OVX rats. Bone mineral content (BMC) and density were significantly decreased in both the lumbar vertebral body and femur of Cd-OVX rats. Ultimate compressive load in the lumbar body and bending load in the midfemur were significantly lower in Cd-OVX rats than in Cont-OVX rats but the differences were not demonstrated when the values were corrected for BMC. Structural moduli in the lumbar vertebral body and the midfemur were not different between the two groups. Cd-OVX rats showed significant decreases in the trabecular bone volume and trabecular number with increased values in the indices of bone formation and resorption in the lumbar vertebral body cancellous bone in comparison with Cont-OVX rats. In the midfemur, Cd-OVX rats had significantly smaller cortical bone area than Cont-OVX rats but the moment of inertia was identical between the two groups. The indices of bone formation and resorption at endocortical surface of the midfemur were significantly increased in Cd-OVX rats over those in Cont-OVX rats, whereas the indices of bone formation at the periosteal surface were not different between the two groups. These data suggested that chronic Cd exposure exacerbated the uncoupling between bone formation and resorption in ovariectomized rats, which resulted in the osteopenia, structural changes of the bone, and decreased mechanical strength in ovariectomized rats with chronic Cd exposure. PMID- 10799338 TI - A comparison of apoptosis and necrosis induced by hepatotoxins in HepG2 cells. AB - 7H-Dibenzo[c,g]carbazole (DBC), an N-heterocyclic aromatic hydrocarbon, is cytotoxic and carcinogenic in rodent liver. While DBC leads to necrotic lesions in the liver, the induction of apoptosis by DBC has not been investigated. The focus of this study was to determine the degree to which apoptosis and necrosis contributed to DBC cytotoxicity in a human hepatoma cell line (HepG2). To determine if these effects were unique to DBC, the results were compared to another hepatotoxin, aflatoxin B(1) (AFB(1)). DBC produced a distinct biphasic LDH release curve within 24 h of exposure. During the same time period lower concentrations of DBC (<10 microM) induced the formation of DBC-DNA adducts and increased p53 protein levels followed by apoptotic cell death. However, increasing the concentration of DBC to 80 microM led to lower DNA adduct and p53 protein levels. At this concentration, intracellular ATP levels were rapidly depleted followed by cell swelling and loss of membrane integrity consistent with necrotic cell death. In contrast to DBC, a biphasic LDH release curve was not observed for AFB(1). Instead, AFB(1) induced a concentration-dependent increase in apoptosis that reached two- to threefold higher levels than DBC. These results suggest that differences exist in the extent and type of cell death induced by DBC and AFB(1) at equimolar concentrations. Apoptosis and necrosis result from low and high concentrations of DBC, respectively, and may be dependent upon intracellular ATP levels. PMID- 10799337 TI - Diflubenzuron, a benzoyl-urea insecticide, is a potent inhibitor of TCDD-induced CYP1A1 expression in HepG2 cells. AB - Diflubenzuron (DFB) belongs to a group of compounds called benzoyphenyl ureas acting as chitin synthesis inhibitors, which also inhibit growth of B16 murine melanomas. The present study was designed to investigate the effect of this insecticide, on CYP1A1 expression and induction in human hepatoma cells HepG2. Treatment of HepG2 cells over 72 h with noncytotoxic concentrations of DFB resulted in a strong dose-dependent decrease in constitutive ethoxyresorufin-O deethylase activity. Moreover, DFB significantly decreased CYP1A1 induction by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) after 24 h exposure, as demonstrated by ethoxyresorufin-O-deethylase (EROD) activity and Northern blot analysis. Additional studies were performed both on parental HepG2 cells and HepG2-241c.1, which were stably transfected with the chloramphenicol acetyltransferase (CAT) reporter gene, cloned under the control of the human CYP1A1 promoter (-1140 to +59). Ribonuclease protection assays (RPA) analysis clearly demonstrated an inhibition of CYP1A1 transcription in both cell lines. Surprisingly, in corresponding experiments using 3-methylcholanthrene (3-MC) as a CYP1A1 inducer, DFB was less effective. Finally, in competitive binding studies using a 9S enriched fraction of HepG2 cytosol, DFB was capable of displacing [(3)H]-2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) from its Ah receptor binding site. Taken together, these results support the involvement of a transcriptional mechanism in the inhibition of CYP1A1 expression in HepG2 cells by DFB, possibly via an Ah receptor antagonism. PMID- 10799339 TI - Cadmium induces c-myc, p53, and c-jun expression in normal human prostate epithelial cells as a prelude to apoptosis. AB - Cadmium is a suspected human prostatic carcinogen shown to induce prostatic tumors and proliferative lesions in rats. The carcinogenic mechanism of cadmium is unknown, but its poor mutagenicity points toward an epigenetic mechanism. Here we studied the effect of cadmium on genes involved in growth regulation of prostate epithelial cell using the human prostate epithelial cell line RWPE-1, which is immortalized but not transformed and is androgen-responsive. Treatment with 10 microM cadmium resulted in transient increases in c-myc and p53 mRNA levels that peaked at 2-fold and 1.4-fold, respectively, compared to control after 2 h. In contrast, c-jun mRNA levels were increased >3-fold after 2, 4, and 6 h and 20-fold after 24 h. DNA synthesis decreased after 24 h of cadmium exposure. Further study revealed a significant increase in apoptosis after 48 h of cadmium exposure. However, approximately 35% of the cells were still viable and appeared normal, indicating this subpopulation was more resistant to cadmium. Furthermore, these resistant cells had 2.5-fold more metallothionein than untreated control cells. This suggests that cadmium could act to select for apoptotic-defective cells in vivo, thereby increasing the likelihood of tumor formation. This work represents the first description of cadmium affecting oncogene expression in a human cell model of a potential in vivo target site of cadmium carcinogenesis. PMID- 10799340 TI - Stress does not enable pyridostigmine to inhibit brain cholinesterase after parenteral administration. AB - The peripherally acting cholinesterase inhibitor pyridostigmine was widely used during the Gulf War as a pretreatment against possible chemical warfare attack. Following consistent reports on long-term illness among Gulf War veterans, pyridostigmine was examined for its possible long-term effects. These effects were suggested to be induced by the combination of pyridostigmine administration and stress exposure that allowed this quaternary compound to enter the brain through stress induced changes in blood-brain barrier (BBB) permeability. Recently, pyridostigmine administration was demonstrated to inhibit brain cholinesterase following acute stress in mice. However, the effect was not replicated under similar conditions in guinea pigs. Because of the significant implication of these findings, we tested brain cholinesterase (ChE) inhibition following the administration of pyridostigmine, or the tertiary carbamate physostigmine, with or without stress in mice. Different experiments were performed to examine the contribution of gender, age (young and adults), stress (type and intensity), or strain (CD-1 and FVB/n) parameters. No inhibition of brain ChE was detected in any of these experiments. At the same time, physostigmine induced the expected decrease in brain ChE in all the experiments. Thus, we could not replicate the findings that suggest pyridostigmine can affect brain cholinesterase following stress. PMID- 10799341 TI - Ethylbenzene induces microsomal oxygen free radical generation: antibody-directed characterization of the responsible cytochrome P450 enzymes. AB - Small aromatic hydrocarbons cause changes in oxidative metabolism by modulating the levels of cytochrome P450 enzymes, with the changes in these enzymes being responsible for qualitative changes in aromatic hydrocarbon metabolism. The goal of this study was to determine if exposure to the small alkylbenzene ethylbenzene (EB) leads to an increase in hepatic free radical production. Male F344 rats were treated with ip injections of EB (10 mmol/kg) and compared to corn oil controls. Hepatic free radical production was examined by measuring the conversion of 2',7' dichlorofluorescin diacetate (DCFH-DA) to its fluorescent product 2',7' dichlorofluorescein (DCF). A significant elevation of fluorescent DCF production was observed after treatment with EB, despite the lack of effect on overall cytochrome P450 levels. This process was shown to be inhibitable by metyrapone, an inhibitor of P450. DCF production was also inhibited by catalase, suggesting that hydrogen peroxide (H(2)O(2)) is one of the reactive oxygen intermediates involved in EB-mediated reactive oxygen species (ROS) formation. Interestingly, superoxide dismutase (SOD) did not inhibit DCF production in corn oil-treated rats but was an effective inhibitor in the EB-treated groups. In an effort to determine if the increase in ROS production was related to changes in specific P450 enzymes, DCF production was measured in the presence of anti-CYP2B, anti CYP2C11, anti-CYP2E1, and anti-CYP3A2 inhibitory antibodies. Anti-CYP2B antibodies inhibited DCF production in EB-treated, but not corn oil groups, which is consistent with the low constitutive levels of this enzyme and its induction by EB. The data also demonstrate that CYP2B contributes to ROS production. Anti CYP2C11 did not influence DCF production in either group. ROS formation in corn oil-treated rats as well as in ethylbenzene-treated rats was also inhibited with antibodies to anti-CYP2E1 and anti-CYP3A2. These results suggest that CYP2C11 does not appear to influence free radical production and that the increase in free radical production in EB treated rats is consistent with the EB-mediated elevation of CYP2B, CYP 2E1, and CYP3A2. Such alterations in free radical generation in response to hydrocarbon treatment may contribute to the toxicity of these compounds. PMID- 10799342 TI - Induction of rat hepatic aryl sulfotransferase (SULT1A1) gene expression by triamcinolone acetonide: impact on minoxidil-mediated hypotension. AB - The hypotensive agent minoxidil (6-imino-1, 2-dihydro-1-hydroxy-2-imino-4 piperidinopyrimidine) depends upon aryl sulfotransferase (SULT1)-catalyzed sulfation for its bioactivation. Previous reports suggest that glucocorticoids induce class-specific SULT1 and isoform-specific SULT1A1 gene expression in rat liver. In the present study, rats were treated with the glucocorticoid triamcinolone acetonide (TA, 5 mg/kg/day i.p. x 3 days) or its vehicle, 2% Tween 20, prior to minoxidil, and subsequent effects on mean arterial pressure (MAP), heart rate (HR), and hepatic SULT1 gene expression were characterized. Minoxidil treatment (1.5 mg/kg) resulted in a steady decline in MAP values of 16.3 to 18.6% relative to basal control levels at 35 to 60 min following minoxidil injection. Pentachlorophenol (PCP, 40 micromol/kg i.p.), an inhibitor of SULT1 enzyme activity, effectively ablated the hypotensive effects of minoxidil. By contrast, pretreatment with TA significantly enhanced minoxidil-induced hypotension. Relative to vehicle-treated controls, TA-treated rats displayed a steeper rate of decline in MAP and more profound levels of hypotension with decreases in MAP following minoxidil administration of 27.8%. TA also produced significant increases in hepatic SULT1 mRNA expression (of 271%) and SULT1A1 immunoreactive protein levels (of 273%), relative to vehicle-treated controls. These results provide physiological evidence to support the biological relevance of SULT1A1 induction by glucocorticoids. The data indicate that steroid treatment induces SULT1A1 gene expression and, as a consequence, accentuates the hypotensive effects of minoxidil. PMID- 10799343 TI - Possible role of caspase-3 inhibition in cadmium-induced blockage of apoptosis. AB - Cadmium (Cd) and chromium (Cr) are human carcinogens. Cr(VI) is taken up into cells and reduced by cellular reductants to the potential DNA damaging species Cr(V), (IV), and (III). Reactive oxygen species and carbon-based radicals may also be produced during Cr reduction. We previously found that Cd blocks Cr induced apoptosis, which could allow a larger proportion of genetically damaged cells to escape and become transformed. This study helped define the mechanisms of Cd-induced suppression of apoptosis. Chinese hamster ovary (CHO K1-BH4) cells were treated with either Cd (5-20 microM), Cr(VI) (350 microM), or Cd (5-20 microM) plus Cr(VI) (350 microM) for 3 h and then cultured in metal-free media for an additional 48 h at which time DNA was extracted or nuclei were examined to determine apoptosis. Cd markedly reduced Cr-induced DNA fragmentation and reduced the number of Cr-induced apoptotic cell nuclei to control levels. Additional study investigated the biokinetics and cellular metabolism of Cr. Cd did not alter the cellular Cr accumulation and there were no differences in the levels of reduced glutathione, a compound possibly important in Cr reduction and reflective of the cellular reducing environment. The antiapoptotic effect of Cd was not due to diminished cellular reduction of Cr(VI) as assessed by electron-spin resonance determination of the levels of Cr(V). Thus, Cd suppression of Cr-induced apoptosis is not based on altered Cr toxicokinetics or metabolism. In addition to Cr, Cd also inhibited apoptosis induced by hygromycin B and actinomycin D. Cd was a very effective inhibitor of caspase-3 activity, a central mediator of apoptosis, with nontoxic levels of Cd resulting in up to approximately 60% inhibition. These results indicate that Cd may have a generalized inhibitory effect on apoptosis, possibly by inhibiting caspase-3. Inhibition of apoptosis by Cd may allow a greater portion of genetically damaged cells to survive, or give selective growth advantages, and has implications as a potential nongenotoxic mechanism of Cd carcinogenesis. PMID- 10799345 TI - Reasonable explanation for both the antiandrogenic mechanism of DBP and DBP induced Leydig cell hyperplasia in prenatally DBP-treated rats. PMID- 10799344 TI - Determining relative estrogenicity by quantifying vitellogenin induction in rainbow trout liver slices. AB - Precision cut tissue slices, by modeling the entire organ, are a valuable tool for studying protein induction or inhibition by test chemicals. This manuscript describes parameters to quantify relative estrogenicity of chemicals in rainbow trout liver slices by measuring vitellogenin (Vg) induction, a well-characterized biomarker of estrogen receptor signal transduction. Hank's medium (phenol-red free) supplemented with Hepes, sodium bicarbonate, and 1% bovine serum albumin was utilized. The experimental parameters were optimized using 1000 nM 17beta estradiol, a potent estrogen in rainbow trout that induces Vg production in vivo. The addition of trout serum and retention of the media was essential, probably to allow for the accumulation of Vg in the slices and media. Histological examination and ATP analyses indicated no toxicity in control or 17beta-estradiol treated liver slices after 120 h. Induction was 4-fold greater with 25% serum containing media compared to media with 10% serum. We observed Vg induction as great as 500-fold over controls at 96 h in liver slices and media containing 25% serum and 1000 nM 17beta-estradiol. Controls without 17beta-estradiol, incubated in media with 10 or 25% serum, exhibited no detectable Vg production, indicating that the induction seen was not from the media or serum. We observed that 48 h was required for significant Vg induction in the media and liver slices. Maximum induction in slices occurred at 96 h, whereas media Vg levels continued to increase to 120 h, suggesting a time delay between Vg production and excretion by the liver. The feasibility of this model to detect weak environmental estrogens was determined with 0-250 microM o,p'DDE and bisphenol A. Both compounds induced Vg in this model with EC50 values of 10(4) and 2x10(5) higher than E(2), respectively. Our results indicate the importance of media, serum, and time selection for optimal Vg induction. This model allows for the determination of relative estrogenicity of chemicals in a controlled in vitro system while utilizing the advantages of precision cut slice technology. PMID- 10799346 TI - Chemical index for volume 164 PMID- 10799347 TI - Pharmacodynamic effects of valerian and hops extract combination (Ze 91019) on the quantitative-topographical EEG in healthy volunteers. AB - The aim of this investigation was to objectify the pharmacodynamic effects of different dosages of a commercially available plant extract mixture of valerian and hops by means of the quantitative topographical EEG (qEEG) in healthy young adults in comparison to placebo. Two different dosages were applied in two single blind, cross-over designed observation trials in 12 healthy volunteers (1st dosage: 500 mg valerian and 120 mg hops, versus placebo, first clinical trial; 2nd dosage: 1500 mg valerian and 360 mg hops, versus placebo, second clinical trial). QEEG was recorded bipolarly from 17 surface electrodes according to the 10:20 system and analysed using the Fast Fourier Transformation prior to, 1, 2 and 4 hours after drug intake in the recording conditions eyes open, eyes closed and under mental demand. The EEG-spectra were cut into six frequency bands. Both resting conditions (eyes open and eyes closed) were analysed together. After application of the low dosage qEEG power changes remained more or less within placebo range following the normal circadian rhythmics, except for a tendentious reduction of alpha- and beta1-power 4 h after drug intake. The high dosage led to power increases in delta, decreases in alpha and a weak decrease in beta-power. Under mental performance only weak differences to placebo were seen which are not discussed here. In the CPT (completion of complicated additions and subtractions) the concentration and performance capability were hardly influenced. However, a minimal increase of mean answer time and mean OK time (time for correct answers) was observed 4 hours after intake of 2 dragees and 1 hour after 6 dragees of valerian and hops mixture with more pronounced changes after the low dosage than the high one. In summary, the quantitative topographical EEG was able to show slight, but clear visible effects on the CNS especially after intake of the high dosage of valerian-hops mixture Ze 91019 indicating reproducible pharmacodynamic responses of the target organ. PMID- 10799348 TI - Fibroblast growth factor cardioprotection against ischemia-reperfusion injury may involve K+ ATP channels. AB - We investigated whether the adenosine triphosphate (ATP)-sensitive K+ (K+ATP) channel that was implicated in the ischemic preconditioning (I-PC) phenomenon, has a role in the cardioprotective effects of fibroblast growth factors (FGFs). For this purpose, we administered glibenclamide, a specific K+ATP channel blocker, before acidic fibroblast growth factor (aFGF, FGF-1) treatment, in rat heart subjected to left ventricular ischemia for 20 minutes followed by reperfusion for 24 hours. Creatine kinase (CK) activity was analyzed in myocardial tissue to assess the degree of cardiac injury. FGF-1 treatment markedly maintains CK activity. This cardioprotective effect of FGF-1 was blocked by glibenclamide. As shown by ultrastructural data, Ca2+ overload and associated cardiomyocyte alterations shown in glibenclamide-treated rats were not observed in specimens from the FGF-1 group. These findings suggest that FGF serves as an effector in I-PC and support a clinical interest of these proteins for increasing myocardial ischemic tolerance. PMID- 10799349 TI - Expansion of CD60 helper lymphocytes detected in peripheral lymphocytes of HIV-1 infected individuals is not paralleled in lymph nodes. AB - BACKGROUND AND OBJECTIVE: A significant expansion of CD8 cells with capability of Th2 type helper function had been observed in hemophiliacs with HIV infection. These cells were characterised by the surface co-expression of CD8 and CD60 antigen. Our objective was to investigate this lymphocyte subset in relation to other subsets in homosexuals and drug users in two compartments: blood and lymph nodes. Blood and lymph nodes from not HIV-infected persons served as control. RESULTS: CD8+CD60+ cells were expanded in perpheral blood of HIV - infected patients as compared to age matched controls (10.0 versus 4.1%, p <0.05). This difference was not observed in lymph node cell suspensions (6.2 vs. 4.3% of all lymph node cells; p = n.s.). The CD 4/CD8 ratio was significantly less impaired in lymph nodes than in blood (2.27 vs. 0.83; p <0.05). Cytotoxic T cells were more abundant in the lymph nodes of patients with early stage HIV disease when compared two late stage patients (4.3 vs 2.1%; p <0.05). Immunohistochemistry on frozen lymph node cuts showed presence of CD60 cells mainly in the interfollicular and paracortical area. In 3 of 10 HIV infected patients these cells were also found in the germinal centers. In controls no CD60 cells were detected in the follicles. Numbers and percentages of CD60 cellls and CD8+CD60+ cells in blood and in lymph nodes did not correlate with HIV - stage, CD4 count or plasma viral load. No correlation with lymph node viral load was seen. CONCLUSIONS: Our data confirm that like in hemophiliacs expansion of CD8+CD60+ is also found in the blood of other HIV risk groups and seems not to be specific for hemophiliacs. However, the higher percentage in peripheral blood is not paralleled in lymph nodes. Redistribution phenomena seem to be the most plausible explanation. According to these data, a major impact of CD8+CD60+ cells in the immunopathogenesis of HIV infection does not seem likely. PMID- 10799350 TI - Ocular massage in a case of central retinal artery occlusion the successful treatment of a hitherto undescribed type of embolism. AB - BACKGROUND: The pathogenesis of central retinal artery occlusions (CRAO) varies, depending upon the underlying disease. An embolic origin of an occlusion often occurs. PATIENT AND METHODS: A 50-year-old man with an occlusion of the left internal carotid artery was examined because of a left central retinal artery occlusion. Fluorescein angiography revealed that in no blood vessel could any circulation of blood be demonstrated. The slight vascular filling with dye mainly came from collateral circulation at the disc. RESULTS: Ocular massage was carried out. After a delay of several minutes, the vessels of the fundus became increasingly filled with blood. The patient noticed continuing recovery in the vision of his left eye. During fluorescein angiography, carried out one day later, white embolic clots appeared in the arteries of the papilla, some of them moving with the velocity of the blood flow through the retinal vessels and reaching the periphery of the retina and immediately disappearing. Others moved more slowly. This was seen repeatedly over several minutes. CONCLUSIONS: In this patient we have recorded in a fluorescein angiogram bright boluses visible in the blood of the retinal arteries. We suggest that conglomerations of blood cells can also cause an obstruction of blood flow. The observation of this kind of bright clots (boluses) visible in the blood of the retinal arteries we did not find described in the literature. In such a situation ocular massage is extremely helpful. Therefore, ocular massage should be carried out in every patient with CRAO. PMID- 10799351 TI - Octreotide in the prevention of pancreatic damage induced by endoscopic sphincterotomy. AB - Somatostatin and the long acting analogue octreotide have been proposed as a therapeutic agent in acute pancreatitis and for the prophylaxis of pancreatic damage by ERCP and EST for their ability to reduce exocrine pancreatic secretion. However, clinical trials could not show significant beneficial effects in acute pancreatitis and ERCP. In patients undergoing EST, data remained controversial, most authors describing positive effects of prophylaxis. In this study we investigated the use of octreotide prophylaxis to reduce EST-induced pancreatic damage in a randomised, double blind trial. 94 consecutive ERCP/EST-patients were randomised to receive either octreotide 200 microgram s.c. or placebo 3 times daily, starting the night before endoscopic procedures. In 59 patients EST was performed. Blood samples were collected before and 40 min, 2 hrs, 6 hrs, 24 hrs, 48 hrs and 72 hrs after the endoscopic procedures. Samples were analysed for pancreatic serum enzymes, acute phase proteins and blood counts. A clinical pain score was investigated. Post-EST-pancreatitis (amylase > 3x upper limit and persistent abdominal pain) was diagnosed in 3 patients in the treatment group, in 4 patients in the placebo group. There were no significant differences in the time-courses of serum enzymes or acute phase proteins in-between the groups, nor in the pain-score. According to these data, prophylactic octreotide application does not prevent acute pancreatic damage induced by endoscopic sphincterotomy. PMID- 10799352 TI - Proof of efficacy of Kamillosan(R) cream in atopic eczema. AB - Kamillosan(R) cream contains chamomile extract as active principle manufactured from the chamomile sort Manzana which is rich in active principles and has been proved not to exhibit a chamomile-related allergen potential. For this reason Kamillosan(R) cream is suited for local therapy of atopic eczema. In a partially double-blind, randomized study carried out as a half-side comparison, Kamillosan(R) cream was tested vs. 0.5% hydrocortisone cream and the vehicle cream as placebo in patients suffering from medium-degree atopic eczema. After a 2-week treatment Kamillosan(R) cream showed a mild superiority towards 0.5% hydrocortisone and a marginal difference as compared to placebo. PMID- 10799353 TI - Treatment of menstrual migraine with prostaglandin synthesis inhibitor mefenamic acid: double-blind study with placebo. AB - The therapeutic effect of mefenamic acid, prostaglandin synthesis inhibitor, on pain of acute menstrual migraine and at the following days during menstrual bleeding period was studied and compared with placebo. 24 patients, 18 to 35 years old, with menstrual migraine were entered for study. They had regular menstrual cycles and they had been diagnosed as experiencing menstrual migraine without aura for more than one year. The patients were treated for 2 consecutive menstrual cycles, one cycle with 500 mg mefenamic acid and one cycle with placebo. Each drug was given at beginning of complaint and similar dose was repeated 8 hourly at following days during the menstrual bleeding period (Total dosage used 1500 mg per day). The use of medication was double blind. Pain intensity was rated by means of a 4 points scale and functional disability was rated from 0 to 3. Results showed that 79.16% of the patients showed significant pain relief with mefenamic acid as compared to 16.6% with placebo. The mean pain score of the mefenamic acid treated attacks decreased significantly from 2.46 +/- 0.5 to 0.62 +/- 1.0 at 2hr postdose. 83.3% of patients treated with mefenamic acid was able to function with or without little effort whereas 12.4% restored their activities with placebo. All the patients (100%) who showed significant initial responses to placebo experienced headache recurrence as compared to 26.3% with mefenamic acid. When considering mean pain scores, percentage of patients with pain free at 2h postdose, percentage of patients required rescue treatment, percentage of patients with headache recurrence and percentages of patients restored full activities, mefenamic acid is significantly superior to placebo in treatment of acute menstrual migraine. It might be concluded that mefenamic acid is safe and effective for treatment of acute menstrual migraine. PMID- 10799354 TI - UV-ozone ashing of cells and tissues for spatially resolved trace element analysis. AB - UV/ozone ashing of thin tissue sections and cell cultures is a simple technique to enhance relative elemental concentrations, while maintaining their spatial location at the sub-micron level. This approach may enhance the capability of spatially resolved analysis techniques to detect the distribution of trace elements in biological matrices. We present results from light microscopy and x ray spectromicroscopy studies of tissues and cells demonstrating that the micro structure is very well conserved. We show the signal enhancement resulting from the removal of carbon, which allows otherwise undetectable gadolinium to be mapped in cancer tissue for a novel neutron capture therapy. PMID- 10799355 TI - Current developments and future prospects for HIV gene therapy using interfering RNA-based strategies. AB - Acquired immunodeficiency syndrome (AIDS) is a slow, progressive, degenerative disease of the human immune system, ultimately leading to premature death of the patient. This disease is primarily caused by human immunodeficiency virus type-1 (HIV-1). The major targets of HIV infection are blood cells, namely lymphocytes and macrophages. While the immune response fails to eliminate the infected cells, the virus continues to spread. The purpose of HIV gene therapy is to provide "anti-HIV" genes to cells that are susceptible to HIV infection. Anti-HIV genes may be designed to express RNAs or proteins that interfere with the function of viral or cellular RNA(s)/protein(s), thereby inhibiting virus replication. Whereas interfering proteins may be cytotoxic and/or immunogenic, interfering RNAs are not. Interfering protein-based strategies requiring inducible gene expression (under the control of HIV regulatory proteins) can only be designed to block steps subsequent to the viral regulatory protein production. In contrast, interfering RNAs can be produced in a constitutive manner, which further enhances their antiviral activity and allows one to design strategies to inhibit virus replication before viral regulatory protein production occurs. Thus, interfering RNAs are of particular interest and are the focus of this review. Genes expressing interfering RNAs were designed to inhibit syncytium formation to prevent the death of the gene-modified cells. Strategies may also be developed to prevent gene-modified cells from becoming infected by HIV or from supporting HIV replication. Genes expressing interfering RNAs have been designed to inhibit HIV 1 entry and to cleave the incoming virion RNA, thus blocking virus replication before provirus DNA synthesis can be completed. A number of genes were also designed to express interfering RNAs that inhibit HIV replication at a post integration step, by inhibiting the function of HIV RNAs or proteins produced in the infected cell. Also in development are anti-HIV genes that produce RNAs that would not only inhibit HIV replication in the gene-modified cell, but also prevent HIV RNA packaging and/or reverse transcription such that the progeny virus produced would be non-infectious. Further refinements to these strategies may lead to the development of "self-propagating" anti-HIV genes. These genes would express interfering RNAs that not only inhibit virus replication in the cell and prevent HIV RNA packaging and/or reverse transcription in the progeny virus, but also make use of the HIV itself to deliver the anti-HIV gene(s) to other cells. Thus, more and more cells susceptible to HIV infection would become resistant. Such "self-propagation" of anti-HIV-1 genes would only occur in cells that are susceptible to HIV infection, and would continue to take place for as long as HIV exists in the body. PMID- 10799356 TI - Cell cycle control in growth plate chondrocytes. AB - Growth of endochondral bones is regulated by the coordinated proliferation and differentiation of chondrocytes in the epiphyseal growth plates. Many skeletal diseases are caused by pathogenic disruptions of these two processes. While the intracellular mechanisms regulating chondrocyte proliferation and differentiation are poorly understood, recent evidence from studies using genetically altered mice and from experiments in cultured chondrocytes point to a prominent role of cell cycle proteins in this context. This article summarizes our current understanding of the expression, regulation, and function of cell cycle genes in chondrocytes. PMID- 10799357 TI - Potential nuclease-based strategies for HIV gene therapy. AB - Gene therapy for HIV (human immunodeficiency virus) involves the introduction of a therapeutic gene into the infected individual for the purposes of reducing viral load and ultimately reconstituting a healthy immune system. Clinical trials for HIV gene therapy have not yet reported therapeutic benefit. In addition to improving the efficiency of gene delivery and the maintenance of gene expression, better therapeutic genes must be designed before this therapy becomes available to patients. A new class of therapeutic genes expressing nucleases may be designed. These nucleases may be classified into three categories based on their mode of action: (i) 'targeted nucleases' for specifically cleaving HIV RNA within the cell, (ii) 'colocalized nucleases' for cleaving HIV genomic DNA or RNA present within the cell or progeny virus, and (iii) 'cytotoxic nucleases' for conferring selective toxicity to HIV-infected cells. The focus of this review is on the design and application of these nucleases for HIV gene therapy. PMID- 10799358 TI - Protein oxidation and age-dependent alterations in calcium homeostasis. AB - Alterations in the capacity to maintain normal calcium homeostasis have been suggested to underlie the reduced cellular function characteristic of the aging process, and to predispose the senescent organism to a host of diverse pathologies including cancer, heart disease, and a range of muscle and neurodegenerative diseases. Therefore, critical to the eventual treatment of many age-related diseases has been the identification of both post-translational modifications and the underlying structural changes that result in an age-related decline in the function of critical calcium regulatory proteins. In brain, multiple methionines within the calcium signaling protein calmodulin (CaM) are oxidized to their corresponding methionine sulfoxides during aging, resulting in an inability to activate a range of target proteins, including the plasma membrane (PM) Ca-ATPase involved in the maintenance of the low intracellular calcium levels necessary for intracellular signaling. Likewise, changes in the transport activity of the PM-Ca-ATPase occur during aging. In muscle, the function of the SERCA2a isoform of the Ca-ATPase within the sarcoplasmic reticulum (SR) declines during aging as a result of the nitration of selected tyrosines. The age-related loss-of-function of these critical calcium regulatory proteins are consistent with observed increases in intracellular calcium levels within senescent cells. A possible regulatory role for these post-translational modifications is discussed, since they have the potential to be reversed following the restoration of normal cellular redox conditions by intracellular repair enzymes that are specific for these post-translational modifications. It is suggested that the reversible oxidation of critical calcium regulatory proteins within excitable cells by reactive oxygen species functions to enhance cellular survival under conditions of oxidative stress by reducing the energy expenditure within excitable cells. Thus, a diminished ability to efficiently generate cellular ATP may ultimately underlie the loss of calcium homeostasis and cellular function during aging. PMID- 10799359 TI - The dietary guideline for sodium: should we shake it up? Yes! AB - The dietary guidelines established under the auspices of public health policy are intended to promote healthy diets in the general public. The current recommendations for sodium intake stem from studies and publications that are older than much of the public they are designed to benefit. The past 2 decades have seen a dramatic increase in our knowledge of nutritional science, particularly our understanding of the role of sodium in blood pressure regulation. With a myriad of data from observational studies and randomized, controlled trials, we have the information to finally put sodium into its correct context in terms of its role in the regulation of blood pressure and hypertension. Not the sole and pervasive dietary villain it was once believed to be, sodium is but one factor in the complex interplay of multiple, inextricably related regulatory systems of which hypertension is the end result. With the data now available concerning dietary sodium, including the minimal and specific blood pressure effects of sodium in normotensive adults and both the benefits and risks of sodium reduction, future public health recommendations can be based on carefully acquired, consistent, and rational science. PMID- 10799360 TI - The dietary guideline for sodium: should we shake it up? No. AB - The current US dietary guideline for sodium is a limit of 2.4 g/d or 6 g NaCl/d. This amount of sodium is far in excess of any physiologic need and is likely an essential though not by itself sufficient primary cause of hypertension as well as a contributor to many other cardiovascular and renal abnormalities. The evidence incriminating the current excessive consumption of sodium derives from epidemiologic, experimental, and interventional data, most of which support a threshold of approximately 100 mmol/d for the harmful effects of sodium to be expressed. Although the current recommendation may not be low enough to go below that threshold, it is an appropriate and attainable goal for now. PMID- 10799361 TI - Vitamin C and myocardial infarction: the heart of the matter. PMID- 10799362 TI - The bioavailability bugaboo. PMID- 10799363 TI - Osteoporosis and vitamin K intake. PMID- 10799364 TI - Skeletal muscle dysfunction in chronic obstructive pulmonary disease and chronic heart failure: underlying mechanisms and therapy perspectives. AB - Low exercise tolerance has a large influence on health status in chronic obstructive pulmonary disease and chronic heart failure. In addition to primary organ dysfunction, impaired skeletal muscle performance is a strong predictor of low exercise capacity. There are striking similarities between both disorders with respect to the muscular alterations underlying the impairment. However, different alterations occur in different muscle types. Histologic and metabolic data show that peripheral muscles undergo a shift from oxidative to glycolytic energy metabolism, whereas the opposite is observed in the diaphragm. These findings are in line with the notion that peripheral and diaphragm muscle are limited mainly by endurance and strength capacity, respectively. In both diseases, muscular impairment is multifactorially determined; hypoxia, oxidative stress, disuse, medication, nutritional depletion, and systemic inflammation may contribute to the observed muscle abnormalities and each factor has its own potential for innovative treatment approaches. PMID- 10799365 TI - Physician-nutrition-specialist track: if we build it, will they come? Intersociety Professional Nutrition Education Consortium. AB - The Intersociety Professional Nutrition Education Consortium (IPNEC) has made substantial progress in its first 2 y. With support from 9 participating nutrition societies and certification organizations and with funding from the National Institutes of Health and several nutrition industry partners, a sustained, functioning consortium has been established. The consortium's 2 principal aims are to establish educational standards for fellowship training of physician nutrition specialists (PNSs) and to create a unified mechanism for certifying physicians who are so trained. Its long-term goals are to increase the pool of PNSs to enable every US medical school to have at least one PNS on its faculty and to surmount obstacles that currently impede the incorporation of nutrition education into the curricula of medical schools and residency programs. The consortium formulated and refined a paradigm for PNSs, conducted a national role delineation survey to define the scope of the discipline of clinical nutrition, and developed a preliminary curriculum template for training PNSs that can be completed in a minimum of 6 mo. IPNEC and its sponsoring societies are strategically positioned to play an important long-term role in nutrition education for physicians. We intend to continue soliciting broad input, especially from directors of fellowship training programs in nutrition and closely related subspecialties; to develop the core content for fellowships in nutrition and related subspecialties; and to initiate a unified PNS certification examination. PMID- 10799366 TI - Mothers' child-feeding practices influence daughters' eating and weight. AB - BACKGROUND: Childhood overweight has increased dramatically, particularly among young girls. Genetic and environmental factors produce the overweight phenotype. Nonshared environments appear to account for a substantial proportion of the population variance in overweight but remain largely unspecified and unmeasured. OBJECTIVE: Our goal was to evaluate the influence of maternal control in feeding, an aspect of nonshared family environment, on daughters' eating and relative weight. DESIGN: Structural equation modeling was used to test models that describe maternal influences on daughters' eating and relative weight. The participants were 197 white, non-Hispanic families with 5-y-old daughters. The mothers' own dietary restraint and their perceptions of their daughters' risk of overweight were used to predict maternal control in feeding, which was used to predict the daughters' eating and weight outcomes. RESULTS: Maternal body mass index was a modest predictor of daughters' relative weight. The addition of the family-environment pathway provided a good fit and showed additional, independent prediction of daughters' relative weight. Mothers' dietary restraint and perceptions of their daughters' risk of overweight predicted maternal child feeding practices, which in turn predicted daughters' eating and relative weight. CONCLUSIONS: Child-specific aspects of the family environment, including mothers' child-feeding practices and perceptions of their daughters' risk of overweight, may represent important, nonshared, environmental influences on daughters' eating and relative weight. The environmental effects noted were modest but comparable in magnitude to the direct association between maternal and child weight, which indicates that measuring family environmental factors can enhance our understanding of the etiology of childhood overweight. PMID- 10799367 TI - Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and in atherosclerotic apolipoprotein E-deficient mice. AB - BACKGROUND: Dietary supplementation with nutrients rich in antioxidants is associated with inhibition of atherogenic modifications to LDL, macrophage foam cell formation, and atherosclerosis. Pomegranates are a source of polyphenols and other antioxidants. OBJECTIVE: We analyzed, in healthy male volunteers and in atherosclerotic apolipoprotein E-deficient (E(0)) mice, the effect of pomegranate juice consumption on lipoprotein oxidation, aggregation, and retention; macrophage atherogenicity; platelet aggregation; and atherosclerosis. DESIGN: Potent antioxidative effects of pomegranate juice against lipid peroxidation in whole plasma and in isolated lipoproteins (HDL and LDL) were assessed in humans and in E(0) mice after pomegranate juice consumption for /=30 g ISP and in non-HDL cholesterol for the groups that consumed >/=40 g ISP. HDL-cholesterol, apo A-I, lipoprotein(a), and triacylglycerol concentrations were not significantly affected by dietary treatment. CONCLUSION: Our findings show that consuming as little as 20 g soy protein/d instead of animal protein for 6 wk reduces concentrations of non-HDL cholesterol and apo B by approximately 2.6% and 2.2%, respectively. 2000;71:-84. PMID- 10799369 TI - Purified eicosapentaenoic and docosahexaenoic acids have differential effects on serum lipids and lipoproteins, LDL particle size, glucose, and insulin in mildly hyperlipidemic men. AB - BACKGROUND: Regular consumption of n-3 fatty acids of marine origin can improve serum lipids and reduce cardiovascular risk. OBJECTIVE: This study aimed to determine whether eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids have differential effects on serum lipids and lipoproteins, glucose, and insulin in humans. DESIGN: In a double-blind, placebo-controlled trial of parallel design, 59 overweight, nonsmoking, mildly hyperlipidemic men were randomly assigned to receive 4 g purified EPA, DHA, or olive oil (placebo) daily while continuing their usual diets for 6 wk. RESULTS: Fifty-six men aged 48.8 +/- 1.1 y completed the study. Relative to those in the olive oil group, triacylglycerols fell by 0.45 +/- 0.15 mmol/L ( approximately 20%; P = 0.003) in the DHA group and by 0.37 +/- 0.14 mmol/L ( approximately 18%; P = 0.012) in the EPA group. Neither EPA nor DHA had any effect on total cholesterol. LDL, HDL, and HDL(2) cholesterol were not affected significantly by EPA, but HDL(3) cholesterol decreased significantly (6.7%; P = 0.032). Although HDL cholesterol was not significantly increased by DHA (3. 1%), HDL(2) cholesterol increased by approximately 29% (P = 0.004). DHA increased LDL cholesterol by 8% (P = 0.019). Adjusted LDL particle size increased by 0.25 +/- 0.08 nm (P = 0.002) with DHA but not with EPA. EPA supplementation increased plasma and platelet phospholipid EPA but reduced DHA. DHA supplementation increased DHA and EPA in plasma and platelet phospholipids. Both EPA and DHA increased fasting insulin significantly. EPA, but not DHA, tended to increase fasting glucose, but not significantly so. CONCLUSIONS: EPA and DHA had differential effects on lipids, fatty acids, and glucose metabolism in overweight men with mild hyperlipidemia. PMID- 10799370 TI - Serum, biliary, and fecal cholesterol and plant sterols in colectomized patients before and during consumption of stanol ester margarine. AB - BACKGROUND: Cholesterol metabolic studies are simplified in colectomized patients because of rapid intestinal passage and reduced bacterial action. OBJECTIVE: Our objective was to study the effect on cholesterol and plant sterol metabolism of feeding a margarine containing stanol ester to 11 colectomized patients. DESIGN: A margarine containing 2 g stanol was consumed for 7-18 d. Serum, biliary, and fecal lipids were measured before and during consumption of the margarine. RESULTS: Serum cholesterol concentrations and the ratio of plant sterol to cholesterol decreased after 1 d of consumption of stanol esters (P < 0.05). After 7 d, serum cholesterol decreased by 16% (P < 0.01), cholesterol absorption efficiency decreased by approximately 40%, and fecal output of cholesterol as neutral sterols (but not as bile acids) increased by 36%. Biliary bile acid composition and the molar percentage of biliary cholesterol were unchanged. Increased ratios of cholesterol precursor sterols in serum and bile indicated enhanced cholesterol synthesis during consumption of stanol esters; the percentage absorption of plant sterols and the ratios of plant sterols to cholesterol decreased, whereas serum and biliary plant stanols and their biliary secretion gradually increased. In feces, 95% of cholesterol and 90% of plant stanols were in unesterified form. CONCLUSIONS: In colectomized patients, effective inhibition of cholesterol absorption and lowering of serum cholesterol concentrations and plant sterol ratios occurs within 1 d of the start of consumption of stanol esters. The composition of major bile lipids is unchanged, indicating that gallstone formation is unlikely. Small amounts of plant stanols are recovered in serum and bile during consumption of stanol esters but effectively are secreted through bile, thereby balancing the intake-induced increase in their absorption. PMID- 10799371 TI - Acute effects of ingestion of black and green tea on lipoprotein oxidation. AB - BACKGROUND: Tea has been associated with a reduced risk of cardiovascular disease. One proposed mechanism of this risk reduction involves inhibition of lipoprotein oxidation in vivo by antioxidant polyphenolic compounds derived from tea. However, controlled interventions uniformly failed to show that ingestion of tea can inhibit LDL oxidation ex vivo. The absence of effects in previous studies may be due to the isolation of LDL particles from polyphenolic compounds that are present in the aqueous phase of serum. OBJECTIVE: The objective of this study was to examine the acute effects of ingestion of black and green tea on ex vivo Cu(2+)-induced lipoprotein oxidation without prior isolation of lipoproteins from serum. DESIGN: The acute effects of 4 hot drinks-green tea and black tea (each at a dose equivalent to 4 standard cups), water matched to the teas for caffeine content, and water-were assessed in 20 healthy men by using a Latin-square design. The lag time to lipoprotein diene formation, slope of the propagation phase of the oxidation curve, and area under the oxidation curve were calculated. Urinary concentrations of 4-O-methylgallic acid were used as a marker of uptake and metabolism of polyphenolic compounds from tea. RESULTS: Significant increases in urinary 4-O-methylgallic acid for black and green tea (P < 0. 0001) were observed. Caffeine did not significantly influence lipoprotein oxidation. Compared with the water control, there was a greater lag time for black tea (5.4 +/- 2.9 min; P = 0.05) that was of borderline significance and a similar trend for green tea (4.4 +/- 2.8 min; P = 0.17). Slope and area under the oxidation curve were not altered. CONCLUSION: Black tea has a mild acute effect on ex vivo lipoprotein oxidation in human serum. 2000;71:-7. PMID- 10799372 TI - Suppression of nocturnal fatty acid concentrations by bedtime carbohydrate supplement in type 2 diabetes: effects on insulin sensitivity, lipids, and glycemic control. AB - BACKGROUND: Bedtime ingestion of slow-release carbohydrates leads to sustained nocturnal fatty acid suppression and improved glucose tolerance in type 2 diabetic patients. OBJECTIVE: This study assessed the effects of 2 different doses of bedtime carbohydrate supplement (BCS) on morning glycemic control and glycated hemoglobin (Hb A(1c)) in type 2 diabetic patients. In addition, the effects of the high-dose BCS on insulin sensitivity and postprandial glucose and triacylglycerol concentrations were assessed. DESIGN: Two BCS doses were studied separately in 7-wk randomized, placebo-controlled, double-blind studies with either a parallel (low-dose BCS; n = 24 patients) or crossover (high-dose BCS; n = 14 patients) design. The effects of the low and high doses (0.30 and 0.55 g uncooked cornstarch/kg body wt, respectively) were compared with those of a starch-free placebo. RESULTS: Compared with the starch-free placebo, the high dose BCS ( approximately 45 g) produced enhanced nocturnal glucose (P < 0.01) and insulin (P < 0.01) concentrations as well as a 32% suppression of fatty acid concentrations (P < 0.01). Moreover, glucose tolerance (P < 0.05) and C-peptide response (P < 0.05) improved after breakfast the next morning. The low-dose BCS ( approximately 25 g) improved fasting blood glucose concentrations (P < 0.05). However, there were no improvements in insulin sensitivity, postprandial triacylglycerol concentrations, or Hb A(1c) after 7 wk. CONCLUSION: Nocturnal fatty acid suppression by BCS improved fasting and postprandial blood glucose concentrations in type 2 diabetic patients the next morning. In contrast, no improvements in insulin sensitivity, postprandial triacylglycerol concentrations, or long-term glycemic control assessed by Hb A(1c) were seen after BCS supplementation. PMID- 10799373 TI - Association between glycated hemoglobin and diet and other lifestyle factors in a nondiabetic population: cross-sectional evaluation of data from the Potsdam cohort of the European Prospective Investigation into Cancer and Nutrition Study. AB - BACKGROUND: Glycation reactions of proteins and other compounds, depending on blood glucose concentrations, have a detrimental effect on health. OBJECTIVE: The association of diet and other lifestyle factors with glycated hemoglobin (Hb A(1c)) values was examined in a nondiabetic population. DESIGN: This was a cross sectional study of 1773 middle-aged men and women. Mean Hb A(1c) values were calculated for categories of diet and lifestyle factors, and odds ratios (ORs) for the highest versus lowest tertiles of Hb A(1c) were determined and compared. RESULTS: The OR of being in the highest Hb A(1c) tertile compared with the lowest increased with greater age [age 40-44 y compared with >60 y: men (OR: 2.86; 95% CI: 1.60, 5.20) and women: (6.11; 3.15, 12.30)] and greater obesity [body mass index (in kg/m(2)) >25 and waist-hip ratio >1.0 in men and >0.8 in women): men (2.80; 1.48, 5.45) and women (1.73; 1.15, 2.61)]. High energy and energy-adjusted saturated fat intakes were associated with increased risk of being in the highest tertile of Hb A(1c) [highest compared with lowest quintile: (1.53; 1.04, 2.26; P for trend = 0.013) and (1. 98; 1.33, 2.95; P for trend = 0.003), respectively]. No significant associations were observed for intakes of carbohydrates, protein, dietary fiber, or beta-carotene; however, some of the associations were nearly significant. Alcohol, vitamin C, and vitamin E intakes were inversely related to risk [highest compared with lowest quintile: (0.56; 0.38, 0.83; P for trend = 0.001), (0.50; 0.33, 0. 74; P for trend = 0.003), and (0.65; 0.43, 0.96; P for trend = 0. 036), respectively]. CONCLUSION: Hb A(1c) values might be modifiable by diet and other lifestyle factors. PMID- 10799374 TI - Arabinoxylan fiber, a byproduct of wheat flour processing, reduces the postprandial glucose response in normoglycemic subjects. AB - BACKGROUND: Arabinoxylan (AX) is the major component of dietary fiber in the cereal grains that make up a large proportion of our diet. However, the physiologic effect of AX is unknown. OBJECTIVE: The objective of this study was to determine whether AX improves postprandial glucose and insulin responses in healthy humans. DESIGN: AX-rich fiber was extracted from the byproduct of wheat flour processing. Three isoenergic breakfasts, comprising bread, margarine, and jam, had 75 g available carbohydrate, 10 g protein, and 14 g fat and contained 0, 6, and 12 g AX-rich fiber, respectively. Fourteen healthy subjects consumed the 3 breakfast meals in random order on 3 mornings >/=3 d apart after an overnight fast. Blood was taken from the subjects at regular intervals over 2 h and was analyzed for glucose and insulin. The palatability of bread containing AX-rich fiber was compared with that of a control bread. RESULTS: Compared with the control meal containing 0 g AX-rich fiber, the peak postprandial glucose concentration after meals containing 6 and 12 g AX-rich fiber was significantly lower (6. 3 +/- 1.3 compared with 7.2 +/- 1.0 mmol/L, P < 0.01; 5.9 +/- 0.9 compared with 7.2 +/- 1.0 mmol/L, P < 0.001, respectively). The incremental area under the curve (IAUC) for glucose was 20.2% (95% CI: 5.8%, 34.7%; P < 0.01) and 41.4% (25.9%, 56.8%; P < 0.001) lower, whereas IAUC for insulin was 17.0% (2.0%, 32.1%; P < 0.05) and 32. 7% (18.8%, 46.6%; P < 0.001) lower, respectively. Bread containing AX-rich fiber was as pala as 50% whole-wheat bread when evaluated with sensory analysis by 30 volunteers. CONCLUSIONS: Postprandial glucose and insulin responses were improved by ingestion of AX-rich fiber. Further research is required to determine whether AX-rich fiber is of benefit to people with type 2 diabetes. PMID- 10799375 TI - Short-term protein and energy supplementation activates nitrogen kinetics and accretion in poorly nourished elderly subjects. AB - BACKGROUND: An increase in protein intake exerts a stimulating effect on protein kinetics in children, young adults, and healthy elderly persons. However, there are few data on the response to such dietary changes in malnourished elderly subjects, despite important medical implications in this population. OBJECTIVE: The objective of this study was to determine the metabolic response to short-term nutritional supplementation in moderately malnourished elderly subjects. DESIGN: The influence of 10 d of supplementation (1.67 MJ/d and 30 g protein/d) on body composition, resting energy expenditure, and whole-body protein kinetics was studied in 17 malnourished elderly patients and 12 healthy young adults. A control group of 6 malnourished elderly patients received no supplementation. RESULTS: Supplemented elderly subjects had a significantly greater fat-free mass gain than did unsupplemented elderly subjects (1.3 and 0.1 kg, respectively; age effect, P < 0.05; diet effect, P < 0.02) and a significantly greater increase in fasting rate of protein synthesis than did young supplemented subjects (0.6 and 0.2 g*kg FFM(-1)*11 h(-1); age effect, P < 0.05). The net protein balance in the supplemented elderly subjects in the fed state was positively correlated with protein intake (r(2) = 0.46) and in the fasted state was negatively correlated with protein intake (r(2) = 0.27). The sum of these regressions is a line with increasingly positive net diurnal protein balance produced by increasing protein intake. CONCLUSION: These data provide evidence of a short-term anabolic response of protein metabolism to dietary supplementation in malnourished elderly patients that is likely to improve muscle strength and functional status. PMID- 10799376 TI - Energy expenditure and free-living physical activity in black and white women: comparison before and after weight loss. AB - BACKGROUND: The prevalence of obesity is higher in black than in white women. Differences in energy economy and physical activity may contribute to this difference. OBJECTIVE: The objective of this study was to compare free-living energy expenditure and physical activity in black and white women before and after weight loss. DESIGN: Participants were 18 white and 14 black women with body mass indexes (in kg/m(2)) between 27 and 30. Diet, without exercise, was used to achieve a weight loss of >/=10 kg and a body mass index <25. After 4 wk of energy balance in overweight and normal-weight states, body composition was assessed by using a 4-compartment model, sleeping and resting energy expenditures were assessed by using a chamber calorimeter, physiologic stress of exercise and exercise economy were measured by using standardized exercise tasks, and daily energy expenditure was assessed by using doubly labeled water. RESULTS: Weight loss averaged 12.8 kg. Sleeping and resting energy expenditures decreased in proportion to changes in body composition. Weight reduction significantly improved physiologic capacity for exercise in both groups of women, making it easier for them to be physically active. Black women had lower body composition adjusted energy requirements than did white women-both before and after weight loss-during sleep (9% lower, 519 kJ/d; P < 0.001), at rest (14% lower, 879 kJ/d; P < 0.001), during exercise (6% lower; P < 0. 05), and as a daily total (9% lower, 862 kJ/d; P < 0.06). By contrast, free-living physical activity was similar between the groups. CONCLUSIONS: Weight-reduced women had metabolic rates appropriate for their body sizes. Black women had lower resting and nonresting energy requirements in both overweight and normal-weight states than did white women and did not compensate with greater physical activity, potentially predisposing them to greater weight regain. PMID- 10799377 TI - Prediction of dietary iron absorption: an algorithm for calculating absorption and bioavailability of dietary iron. AB - BACKGROUND: Dietary iron absorption from a meal is determined by iron status, heme- and nonheme-iron contents, and amounts of various dietary factors that influence iron absorption. Limited information is available about the net effect of these factors. OBJECTIVE: The objective was to develop an algorithm for predicting the effects of factors known to influence heme- and nonheme-iron absorption from meals and diets. DESIGN: The basis for the algorithm was the absorption of iron from a wheat roll (22.1 +/- 0.18%) containing no known inhibitors or enhancers of iron absorption and adjusted to a reference dose absorption of 40%. This basal absorption was multiplied by the expected effect of different amounts of dietary factors known to influence iron absorption: phytate, polyphenols, ascorbic acid, meat, fish and seafood, calcium, egg, soy protein, and alcohol. For each factor, an equation describing the dose-effect relation was developed. Special considerations were made for interactions between individual factors. RESULTS: Good agreement was seen when measurements of iron absorption from 24 complete meals were compared with results from use of the algorithm (r(2) = 0.987) and when mean iron absorption in 31 subjects served a varied whole diet labeled with heme- and nonheme-iron tracers over a period of 5 d was compared with the mean total iron absorption calculated by using the algorithm (P = 0.958). CONCLUSIONS: This algorithm has several applications. It can be used to predict iron absorption from various diets, to estimate the effects expected by dietary modification, and to translate physiologic into dietary iron requirements from different types of diets. PMID- 10799379 TI - Bioavailability of the calcium in fortified soy imitation milk, with some observations on method. AB - BACKGROUND: Calcium-fortified soy milk is growing in popularity, particularly among vegetarians, but the bioavailability of its calcium was not previously known. Additionally, the validity of isotopic labeling methods for fortified liquid products had not been established. OBJECTIVES: The objectives of this study were to compare the bioavailability of the calcium in fortified soy milk with that of calcium in cow milk and to evaluate the method of labeling soy milk for bioavailability testing. DESIGN: A within-subject comparison of extrinsically labeled cow milk with intrinsically and extrinsically labeled soy milks was undertaken in 16 healthy men. In all tests, 300-mg Ca loads were given as a part of a light breakfast after an overnight fast. The milks were physically partitioned into liquid and solid phases to enable evaluation of tracer distribution. RESULTS: Calcium from intrinsically labeled soy milk was absorbed at only 75% the efficiency of calcium from cow milk. Extrinsic labeling of soy milk did not produce uniform tracer distribution throughout the liquid and solid phases and resulted in a 50% overestimate of true absorbability. CONCLUSION: Calcium-fortified soy milk does not constitute a calcium source comparable to cow milk, and extrinsic labeling of such calcium particulate suspensions does not produce the uniform tracer distribution needed for bioavailability testing. Hence, intrinsic labeling of the fortificant is required for such liquid suspensions. PMID- 10799378 TI - Serum 25-hydroxyvitamin D concentrations and related dietary factors in peri- and postmenopausal Japanese women. AB - BACKGROUND: Few studies of vitamin D nutrition in Asian populations have been conducted. OBJECTIVE: The objective was to assess 25-hydroxyvitamin D [25(OH)D] concentrations in healthy elderly Japanese women during the winter and to determine whether 25(OH)D concentrations are associated with lifestyle. DESIGN: We investigated 151 women aged 66.5 +/- 6.7 y (f1.gif" BORDER="0"> +/- SD) living in a rural community in February 1999. Serum 25(OH)D and intact parathyroid hormone were measured by using HPLC and an immunoradiometric assay, respectively. Information on lifestyle factors, including sunshine exposure and the consumption of vitamin D-rich foods, was also obtained through an interview. RESULTS: The mean (+/-SD) 25(OH)D concentration was 59.9 +/- 17.0 nmol/L. Vitamin D insufficiencies (<30 nmol/L) were found in 4.6% of the women, a value lower than that found in white populations. No correlation was found between age and 25(OH)D concentrations (r = 0.004, P = 0.957). The 25(OH)D concentration of subjects who consumed fish frequently (>/=4 times/wk) was 10.1 nmol/L higher (P < 0.001) than that of subjects with a moderate consumption of fish (1-3 times/wk). Additionally, those who did not consume eggs had significantly lower 25(OH)D concentrations than did those who consumed eggs >/=1 time/wk (P < 0.05). CONCLUSIONS: : The nutritional status of vitamin D in Japanese populations seems to be better than that in most Western populations. Frequent fish consumption is believed to help maintain adequate concentrations of serum 25(OH)D in elderly Japanese women during the winter. PMID- 10799380 TI - Intestinal absorption of beta-carotene ingested with a meal rich in sunflower oil or beef tallow: postprandial appearance in triacylglycerol-rich lipoproteins in women. AB - BACKGROUND: Evidence indicates that different types of fat have different effects on the postprandial plasma triacylglycerol response. Therefore, the type of fat may influence the appearance of beta-carotene in postprandial triacylglycerol rich lipoproteins, which is used as an indicator of intestinal beta-carotene absorption. OBJECTIVE: We compared in female subjects the appearance of beta carotene in plasma triacylglycerol-rich lipoproteins after beta-carotene was ingested with a meal containing sunflower oil or beef tallow. DESIGN: Women (n = 11) each ingested 2 different vitamin A-free, fat-rich meals that were supplemented with beta-carotene (47 micromol) and contained equivalent amounts (60 g) of sunflower oil or beef tallow. Blood samples were collected hourly from 0 to 10 h; additional samples were collected at selected intervals until 528 h. In a subgroup of the women (n = 7), plasma chylomicrons and 3 subfractions of VLDLs were separated by cumulative rate ultracentrifugation. RESULTS: The appearance of beta-carotene in chylomicrons and in each VLDL subfraction was lower after ingestion with the meal containing sunflower oil than after ingestion with the meal containing beef tallow (P < 0.03). In chylomicrons, the area under the concentration-versus-time curve (AUC) for beta-carotene was 38.1 +/- 13.6% lower (P < 0.03); in contrast, the AUC for triacylglycerol was higher (P < 0.05) after the sunflower-oil-rich meal than after the beef-tallow-rich meal. CONCLUSIONS: Ingestion of beta-carotene with a meal rich in sunflower oil as compared with a meal rich in beef tallow results in lower appearance of beta carotene and greater appearance of triacylglycerol in triacylglycerol-rich lipoproteins. PMID- 10799381 TI - Vitamin C and the risk of acute myocardial infarction. AB - BACKGROUND: Low-fat soluble-antioxidant status is associated with an increased risk of heart disease. OBJECTIVE: The aim of this study was to examine whether low plasma concentrations of vitamin C confer an independent risk of acute myocardial infarction (AMI). DESIGN: Male patients (n = 180) aged <65 y with a first AMI and without an existing diagnosis of angina (>6 mo) who were admitted within 12 h after onset of symptoms were compared with apparently healthy volunteers (n = 177). Plasma concentrations and dietary intakes of vitamin C were determined during hospitalization and 3 mo later. RESULTS: Compared with the control subjects, the patients had higher total cholesterol and lower HDL cholesterol concentrations and more of them smoked. The relative risk of AMI for the lowest compared with the highest quintile of plasma vitamin C during hospitalization (14.5 and >60.5 micromol/L, respectively) was 8.37 (95% CI: 3.28, 21. 4) after adjustment for classic risk factors. At 3 mo, mean (+/-SEM) plasma vitamin C concentrations in patients had increased significantly, from 19.6 +/- 1.2 to 35.1 +/- 1.9 micromol/L (P < 0. 001) and no longer conferred a risk of AMI [relative risk: 1.02 (95% CI: 0.51, 2.03)]. Habitual dietary vitamin C intake of patients (before AMI) did not differ significantly from that of control subjects. The increase in plasma vitamin C after recovery from the infarction could not be explained by a similarly large increase in dietary vitamin C. CONCLUSIONS: A low plasma concentration of vitamin C was not associated with an increased risk of AMI, irrespective of smoking status. The apparent risk of AMI due to a low plasma vitamin C concentration was distorted by the acute phase response. PMID- 10799382 TI - Amount of fat in the diet affects bioavailability of lutein esters but not of alpha-carotene, beta-carotene, and vitamin E in humans. AB - BACKGROUND: Fat-soluble vitamin E and carotenoids are regarded as being protective against chronic diseases. Little is known about the effect of dietary fat on the bioavailability of these compounds. OBJECTIVE: The objective of this study was to assess the effect of the amount of dietary fat on plasma concentrations of vitamin E and carotenoids after supplementation with these compounds. DESIGN: During two 7-d periods, 4 groups of 14-15 volunteers received daily, with a low-fat hot meal, 1 of 4 different supplements: vitamin E (50 mg), alpha- plus beta-carotene (8 mg), lutein esters (8 mg lutein), or placebo. The supplements were provided in a low- or high-fat spread supplied in random sequence during either of the 2 experimental periods. RESULTS: As anticipated, plasma concentrations of vitamin E, alpha- and beta-carotene, and lutein were significantly higher in the supplemented groups than in the placebo group. The amount of dietary fat consumed with the hot meal (3 or 36 g) did not affect the increases in plasma concentrations of vitamin E (20% increase with the low-fat spread and 23% increase with the high-fat spread) or alpha- and beta-carotene (315% and 139% with the low-fat spread and 226% and 108% with the high-fat spread). The plasma lutein response was higher when lutein esters were consumed with the high-fat spread (207% increase) than with the low-fat spread (88% increase). CONCLUSION: Optimal uptake of vitamin E and alpha- and beta-carotene requires a limited amount of fat whereas the amount of fat required for optimal intestinal uptake of lutein esters is higher. 2000;71:-93. PMID- 10799383 TI - Direct correlation of glutathione and ascorbate and their dependence on age and season in human lymphocytes. AB - BACKGROUND: Endogenous reactive oxygen species appear to contribute to aging and cancer and dietary antioxidants, present in fruit and vegetables, counteract these effects. OBJECTIVE: The objective was to examine the association between intracellular glutathione, ascorbate (vitamin C), and alpha-tocopherol (vitamin E) in human lymphocytes. DESIGN: The study group consisted of 240 healthy nonsmoking volunteers with an approximately equal number of male and female subjects subdivided into 3 age groups: 18-39, 40-59, and >/=60 y). Glutathione, glutathione disulfide, ascorbate, and alpha-tocopherol were measured in lymphocytes by HPLC. RESULTS: The average concentration of antioxidants in lymphocytes was 27 +/- 8 nmol/mg protein for glutathione, 21 +/- 8 nmol/mg protein for ascorbate, and 0.4 +/- 0.2 nmol/mg protein for alpha-tocopherol. There was a strong positive correlation between glutathione and ascorbate (r = 0.62, P < 0.001). No correlation was observed for glutathione and ascorbate with alpha-tocopherol. The concentration of glutathione in lymphocytes was inversely correlated with age (r = -0.19, P < 0.01), as was that of ascorbate (r = -0.22, P < 0.01), with 10-20% lower values in elderly than in young and elderly subjects. The concentrations of glutathione in lymphocytes were as much as 25% higher and those of ascorbate were as much as 38% higher during the summer than during the winter. The seasonal variation of ascorbate in lymphocytes was described by a linear function for age and a periodic sine function for season. CONCLUSION: Glutathione and ascorbate are directly correlated in human lymphocytes. PMID- 10799385 TI - Protein intake and appendicular skeletal muscle mass in older men. PMID- 10799384 TI - Dietary vitamin K intakes are associated with hip fracture but not with bone mineral density in elderly men and women. AB - BACKGROUND: Vitamin K has been associated with bone mineral density (BMD) and risk of hip fracture. The apolipoprotein (apo) E4 allele (APOE*E4) has been associated with bone fracture through a putative effect on vitamin K transport in blood. OBJECTIVE: The objective was to determine the associations between vitamin K intake, apo E genotype, BMD, and hip fracture in a population-based cohort of elderly men and women. DESIGN: Dietary vitamin K intake was assessed with a food frequency questionnaire in 335 men and 553 women (average age: 75.2 y) participating in the Framingham Heart Study in 1988-1989. Incidence of hip fractures was recorded from 1988 to 1995. BMD at the hip, spine, and arm was assessed on 2 separate occasions (1988-1989 and 1992-1993). Comparisons between apo E genotype and BMD were made relative to E4 allele status (at least 1 epsilon4 allele compared with no epsilon4 allele). RESULTS: Individuals in the highest quartile of vitamin K intake (median: 254 microg/d) had a significantly lower fully adjusted relative risk (0.35; 95% CI: 0. 13, 0.94) of hip fracture than did those in the lowest quartile of intake (median: 56 microg/d). There were no associations between vitamin K intake and BMD in either men or women. No association was found between the E4 allele and BMD, and there were no significant interactions between the E4 allele and phylloquinone intake and BMD or hip fracture. CONCLUSIONS: Low vitamin K intakes were associated with an increased incidence of hip fractures in this cohort of elderly men and women. Neither low vitamin K intake nor E4 allele status was associated with low BMD. PMID- 10799387 TI - Lutein supplementation or green vegetables. PMID- 10799389 TI - Use of the term vegetarian. PMID- 10799391 TI - Dietary copper and risk of coronary heart disease. PMID- 10799393 TI - Maternal Nutrition: New Developments and Implications. Proceedings of a symposium. Paris, France, June 11-12, 1998. PMID- 10799394 TI - Physiology of pregnancy and nutrient metabolism. AB - Pregnancy consists of a series of small, continuous physiologic adjustments that affect the metabolism of all nutrients. The adjustments undoubtedly vary widely from woman to woman depending on her prepregnancy nutrition, genetic determinants of fetal size, and maternal lifestyle behavior. Studies of protein and energy metabolism illustrate the potential of adjusting the use of those nutrients to conserve a fetal supply. Adjustments in the metabolism of nitrogenous compounds are in place by the second quarter of pregnancy. During the last quarter of pregnancy, when fetal demands are greatest, those adjustments allow a positive nitrogen retention. The energy requirement of basal metabolism is influenced by maternal prepregnant nutrition and by fetal size. If maternal energy reserves are low at conception, the basal metabolic rate is down-regulated to conserve energy. Also, women having larger babies tend to have greater increases in their basal metabolic rate and lower rates of maternal energy storage. Changes in maternal food and physical activity behaviors during gestation may augment the physiologic adjustments. However, the substantial variability in food intakes and physical activity makes it difficult to show those changes. Thresholds in the capacity to adjust nutrient use to the amount supplied exist for all nutrients. When intakes fall below the threshold, fetal growth and development is affected more than is maternal health. Efforts to achieve good maternal nutritional status preconception as well as throughout gestation best assure a good milieu for fetal growth and development. PMID- 10799395 TI - Energy adaptations in human pregnancy: limits and long-term consequences. AB - The very slow rate of human fetal growth generates a lower incremental energy stress than in any other mammalian species. This creates a situation in which adaptive changes in metabolic rate and in the amount of additional maternal fat stored during gestation can make a profound difference to the overall energy needs of pregnancy. Comparisons of women in affluent and poor countries have recorded mean population energy needs ranging from as high as 520 MJ to as low as -30 MJ per pregnancy. These energy costs are closely correlated with maternal energy status when analyzed both between and within populations, suggesting that they represent functional adaptations that have been selected for their role in protecting fetal growth. Although this metabolic plasticity represents a powerful mechanism for sustaining pregnancy under very marginal nutritional conditions, it must not be construed as a perfect mechanism that obviates the need for optimal nutritional care of pregnant women. The fact that fetal weight represents up to 60% of total pregnancy weight gain in many pregnancies in poor societies (compared with a well-nourished norm of 25%) indicates that the fetus is developing under suboptimal nutritional and physiologic conditions. It has long been recognized that this has immediate consequences for the offspring in terms of increased perinatal mortality. The more recent appreciation that impaired fetal growth may also precipitate longer-term defects in terms of adult susceptibility to noncommunicable and infectious diseases reinforces the view that pregnancy may be the most sensitive period of the life cycle in which nutritional intervention may reap the greatest benefits. PMID- 10799396 TI - Pregnancy weight gain: still controversial. AB - During the 20th century, recommendations for maternal weight gain in pregnancy were controversial, ranging from rigid restriction to encouragement of ample gain. In 1990, the Institute of Medicine (IOM) recommended weight-gain ranges with the primary goal of improving infant birth weight. These guidelines were widely adopted but not universally accepted. Critics have argued that the IOM's recommendations are unlikely to improve perinatal outcomes and may actually increase the risk of negative consequences to both infants and mothers. We systematically reviewed studies that examined fetal and maternal outcomes according to the IOM's weight-gain recommendations in women with a normal prepregnancy weight. These studies showed that pregnancy weight gain within the IOM's recommended ranges is associated with the best outcome for both mothers and infants. However, weight gain in most pregnant women is not within the IOM's ranges. All of the studies reviewed were observational and there is a compelling need to conduct experimental studies to examine interventional strategies to improve maternal weight gain with the objective of optimizing health outcomes. PMID- 10799397 TI - Obesity and pregnancy: complications and cost. AB - The prevalence of obesity is currently rising in developed countries, making pregravid overweight one of the most common high-risk obstetric situations. Although the designs and populations of published studies vary widely, most authors agree that pregravid overweight increases maternal and fetal morbidity. Even moderate overweight is a risk factor for gestational diabetes and hypertensive disorders of pregnancy, and the risk is higher in subjects with overt obesity. Compared with normal weight, maternal overweight is related to a higher risk of cesarean deliveries and a higher incidence of anesthetic and postoperative complications in these deliveries. Low Apgar scores, macrosomia, and neural tube defects are more frequent in infants of obese mothers than in infants of normal-weight mothers. The regional distribution of fat modulates the effects of weight on carbohydrate tolerance, hemodynamic adaptation, and fetal size. Maternal obesity increases perinatal mortality. Long-term complications include worsening of maternal obesity and development of obesity in the infant. The average cost of hospital prenatal and postnatal care is higher for overweight mothers than for normal-weight mothers, and infants of overweight mothers require admission to neonatal intensive care units more often than do infants of normal weight mothers. Preconception counseling, careful prenatal management, tight monitoring of weight gain, and long-term follow-up could minimize the social and economic consequences of pregnancies in overweight women. PMID- 10799398 TI - Protein metabolism in pregnancy. AB - Adaptation to pregnancy involves major changes in maternal metabolism to provide for the growing demands of the conceptus. Although changes in glucose metabolism, and possibly in fatty acid metabolism, occur in parallel with the increasing energy demands of the mother and the fetus, adaptation of protein metabolism appears to be in anticipation of maternal and fetal needs. During pregnancy, there is an excess of maternal nitrogen in the form of lean body mass over that deposited in the fetus and the products of conception; there is also a pregnancy induced hypoaminoacidemia and a diminished amino acid response to protein intake, suggesting an increased uptake of amino acids in the splanchnic compartment. With the use of stable-isotope-labeled tracers, it was shown that there is a decreased rate of urea synthesis during pregnancy that is evident early in gestation. Kinetic studies of leucine metabolism showed no significant change in leucine carbon turnover but a significantly lower rate of leucine nitrogen turnover, suggesting a lower rate of leucine transamination. These data suggest an integral regulation of whole-body protein and nitrogen metabolism starting early in gestation and aimed at conservation and accretion of nitrogen by the mother and the fetus. PMID- 10799399 TI - Carbohydrate and lipid metabolism in pregnancy: normal compared with gestational diabetes mellitus. AB - This article reviews maternal metabolic strategies for accommodating fetal nutrient requirements in normal pregnancy and in gestational diabetes mellitus (GDM). Pregnancy is characterized by a progressive increase in nutrient stimulated insulin responses despite an only minor deterioration in glucose tolerance, consistent with progressive insulin resistance. The hyperinsulinemic euglycemic glucose clamp technique and intravenous-glucose-tolerance test have indicated that insulin action in late normal pregnancy is 50-70% lower than in nonpregnant women. Metabolic adaptations do not fully compensate in GDM and glucose intolerance ensues. GDM may reflect a predisposition to type 2 diabetes or may be an extreme manifestation of metabolic alterations that normally occur in pregnancy. In normal pregnant women, basal endogenous hepatic glucose production (R(a)) was shown to increase by 16-30% to meet the increasing needs of the placenta and fetus. Total gluconeogenesis is increased in late gestation, although the fractional contribution of total gluconeogenesis to R(a), quantified from (2)H enrichment on carbon 5 of glucose (65-85%), does not differ in pregnant women after a 16-h fast. Endogenous hepatic glucose production was shown to remain sensitive to increased insulin concentration in normal pregnancy (96% suppression), but is less sensitive in GDM (80%). Commensurate with the increased rate of glucose appearance, an increased contribution of carbohydrate to oxidative metabolism has been observed in late pregnancy compared with pregravid states. The 24-h respiratory quotient is significantly higher in late pregnancy than postpartum. Recent advances in carbohydrate metabolism during pregnancy suggest that preventive measures should be aimed at improving insulin sensitivity in women predisposed to GDM. Further research is needed to elucidate the mechanisms and consequences of alterations in lipid metabolism during pregnancy. PMID- 10799400 TI - Essential fatty acids in mothers and their neonates. AB - Essential fatty acids (EFAs) and their long-chain polyenes (LCPs) are indispensable for human development and health. Because humans cannot synthesize EFAs and can only ineffectively synthesize LCPs, EFAs need to be consumed as part of the diet. Consequently, the polyunsaturated fatty acid (PUFA) status of the developing fetus depends on that of its mother, as confirmed by the positive relation between maternal PUFA consumption and neonatal PUFA status. Pregnancy is associated with a decrease in the biochemical PUFA status, and normalization after delivery is slow. This is particularly true for docosahexaenoic acid (DHA) because, on the basis of the current habitual diet, birth spacing appeared to be insufficient for the maternal DHA status to normalize completely. Because of the decrease in PUFA status during pregnancy, the neonatal PUFA status may not be optimal. This view is supported by the lower neonatal PUFA status after multiple than after single births. The neonatal PUFA status can be increased by maternal PUFA supplementation during pregnancy. For optimum results, the supplement should contain both n-6 and n-3 PUFAs. The PUFA status of preterm neonates is significantly lower than that of term infants, which is a physiologic condition. Because the neonatal DHA status correlates positively with birth weight, birth length, and head circumference, maternal DHA supplementation during pregnancy may improve the prognosis of preterm infants. In term neonates, maternal linoleic acid consumption correlates negatively with neonatal head circumference. This suggests that the ratio of n-3 to n-6 PUFAs in the maternal diet should be increased. Consumption of trans unsaturated fatty acids appeared to be associated with lower maternal and neonatal PUFA status. Therefore, it seems prudent to minimize the consumption of trans fatty acids during pregnancy. PMID- 10799401 TI - Role of cholesterol in embryonic development. AB - We showed previously that 3 distal inhibitors of cholesterol synthesis are highly teratogenic in rats. AY 9944 and BM 15766 inhibit 7-dehydrocholesterol reductase, which catalyzes the last step of cholesterol synthesis, and triparanol inhibits Delta(24)-dehydrocholesterol reductase, which catalyzes the last step in another pathway. These molecules cause holoprosencephalic brain anomalies. Under certain experimental conditions, other anomalies (of the limbs and male genitalia) are also observed. Assays performed by gas chromatography-mass spectrometry (GC-MS) show hypocholesterolemia and an accumulation of precursors. These data indicate that this animal model can be considered a model of Smith-Lemli-Opitz syndrome. Smith-Lemli-Opitz syndrome is a recessive autosomal genetic disease characterized by malformations (microcephaly, corpus callosum agenesis, holoprosencephaly, and mental retardation), male pseudohermaphroditism, finger anomalies, and failure to thrive. The syndrome has been attributed to a deficit in 7-dehydrocholesterol reductase. As assayed by GC-MS, the sterol status of these patients indicates severe hypocholesterolemia and an accumulation of precursors: 7 dehydrocholesterol, 8-dehydrocholesterol, and oxidized derivatives. The presence of 7-dehydrocholesterol in the serum of patients is pathognomonic of the disease. The developmental gene Shh (sonic hedgehog) plays a key role in brain, limb, and genital development; it was shown recently that the Shh protein has to be covalently linked to cholesterol to be active. This is the first time that a posttranslational function has been attributed to cholesterol. There is an obvious relation between Shh dysfunction and the malformations observed in our experiments and in patients with Smith-Lemli-Opitz syndrome. However, the exact relation remains to be clarified. It is clear, however, that the role of cholesterol in embryonic development must be taken into account. PMID- 10799402 TI - Anemia and iron deficiency: effects on pregnancy outcome. AB - This article reviews current knowledge of the effects of maternal anemia and iron deficiency on pregnancy outcome. A considerable amount of information remains to be learned about the benefits of maternal iron supplementation on the health and iron status of the mother and her child during pregnancy and postpartum. Current knowledge indicates that iron deficiency anemia in pregnancy is a risk factor for preterm delivery and subsequent low birth weight, and possibly for inferior neonatal health. Data are inadequate to determine the extent to which maternal anemia might contribute to maternal mortality. Even for women who enter pregnancy with reasonable iron stores, iron supplements improve iron status during pregnancy and for a considerable length of time postpartum, thus providing some protection against iron deficiency in the subsequent pregnancy. Mounting evidence indicates that maternal iron deficiency in pregnancy reduces fetal iron stores, perhaps well into the first year of life. This deserves further exploration because of the tendency of infants to develop iron deficiency anemia and because of the documented adverse consequences of this condition on infant development. The weight of evidence supports the advisability of routine iron supplementation during pregnancy. PMID- 10799403 TI - Maternal hemoglobin concentration and birth weight. AB - Pregnancy requires additional maternal absorption of iron. Maternal iron status cannot be assessed simply from hemoglobin concentration because pregnancy produces increases in plasma volume and the hemoglobin concentration decreases accordingly. This decrease is greatest in women with large babies or multiple gestations. However, mean corpuscular volume does not change substantially during pregnancy and a hemoglobin concentration <95 g/L in association with a mean corpuscular volume <84 fL probably indicates iron deficiency. Severe anemia (hemoglobin <80 g/L) is associated with the birth of small babies (from both preterm labor and growth restriction), but so is failure of the plasma volume to expand. Hemoglobin concentrations >120 g/L at the end of the second trimester are associated with a 84 fL, should be considered optimal. PMID- 10799404 TI - Effectiveness and strategies of iron supplementation during pregnancy. AB - Iron deficiency continues to be one of the most prevalent single-nutrient deficiencies in the world. Interventions are often designed to prevent the decrease in hemoglobin concentration and the decline in iron stores associated with pregnancy. Although this is believed to be desirable for both the health of the mother and the well-being of the growing fetus, some scientists disagree. Enrichment and fortification of food items, and dietary changes resulting from education interventions, have met with some success in developed countries, but not often in the developing world. A therapeutic approach to iron supplementation, rather than a public health-based approach, is used throughout much of the world but suffers from real, or perceived, problems of compliance. Large doses of iron are most often prescribed and are associated with side effects and with increased oxidative damage. Alternatively, delayed-release preparations and intermittent oral iron supplementation lead to better overall compliance and alleviate side effects. Daily iron intervention provides more protection against a decline in the storage iron pool in pregnant women than does an intermittent schedule, but the latter is generally associated with fewer side effects, better compliance, and possibly a reduction in risk of oxidative damage. An improved cost-benefit ratio associated with a lower-dose oral iron supplement may prove to be quite positive in the future. Currently, no single approach may be universally acceptable, although a moderate iron dosage protocol will likely provide the most benefit to those who require supplemental iron. PMID- 10799405 TI - Folic acid: influence on the outcome of pregnancy. AB - The periconceptional use of folic acid-containing supplements reduces the first occurrence, as well as the recurrence, of neural tube defects. Women of populations in which adverse pregnancy outcomes are prevalent often consume diets that contain a low density of vitamins and minerals, including folate. Folate intake may need to be sustained after complete closure of the neural tube to decrease the risk of other poor pregnancy outcomes. A central feature of embryonic and fetal development is widespread cell division; folate is central because of its role in nucleic acid synthesis. During gestation, marginal folate nutriture can impair cellular growth and replication in the fetus or placenta. Folate deficiency can occur because dietary folate intake is low or because the metabolic requirement for folate is increased by a particular genetic defect or defects. During pregnancy, low concentrations of dietary and circulating folate are associated with increased risks of preterm delivery, infant low birth weight, and fetal growth retardation. A metabolic effect of folate deficiency is an elevation of blood homocysteine. Likewise, the presence of maternal homocysteine concentrations have been associated both with increased habitual spontaneous abortion and pregnancy complications (eg, placental abruption and preeclampsia), which increase the risk of poor pregnancy outcome and of decreased birth weight and gestation duration. PMID- 10799406 TI - New standard for dietary folate intake in pregnant women. AB - The Institute of Medicine Panel for Folate and Other B Vitamins and Choline considered data from population-based and metabolic studies to revise the dietary intake standards for pregnancy. The recommended dietary allowance (RDA) for pregnant women is the average daily dietary intake sufficient to meet the requirements of 97-98% of pregnant women. The RDA is derived from the amount estimated to meet the requirement of half of healthy pregnant women, or the estimated average requirement (EAR). Maintenance of red cell folate was selected as the primary indicator of adequacy of folate status during pregnancy. The dietary folate equivalent (DFE) was used to interpret studies in which folate was provided as a combination of food folate and synthetic folic acid because folic acid is more bioavailable than is food folate. Many population-based studies confirmed that approximately 680 nmol (approximately 300 microg) folic acid/d consumed in conjunction with a low-folate diet prevented folate deficiency in pregnant women. Additional studies showed that 227 nmol (100 microg) folic acid/d was inadequate to maintain normal folate status in a significant percentage of the groups assessed. The EAR was derived by adding the DFE of this quantity [454 nmol (200 microg)/d] to the EAR for nonpregnant women [725 nmol (320 microg)/d] to provide an EAR of 1178 nmol (520 microg)/d. The RDA of 1362 nmol (600 microg) DFE/d was derived by multiplying the EAR by 1.2 to account for an estimated 10% CV. Data from the metabolic studies support an RDA of 1362 nmol (600 microg) DFE/d on the basis of the maintenance of normal red cell folate concentrations and agree with the findings from the population studies that 1362 nmol DFE/d is adequate to maintain normal folate status in pregnant women. PMID- 10799407 TI - Response of red blood cell folate to intervention: implications for folate recommendations for the prevention of neural tube defects. AB - Committees worldwide have set almost identical folate recommendations for the prevention of the first occurrence of neural tube defects (NTDs). We evaluate these recommendations by reviewing the results of intervention studies that examined the response of red blood cell folate to altered folate intake. Three options are suggested to achieve the extra 400 microg folic acid/d being recommended by the official committees: increased intake of folate-rich foods, dietary folic acid supplementation, and folic acid fortification of food. A significant increase in foods naturally rich in folates was shown to be a relatively ineffective means of increasing red blood cell folate status in women compared with equivalent intakes of folic acid-fortified food, presumably because the synthetic form of the vitamin is more stable and more bioavailable. Although folic acid supplements are highly effective in optimizing folate status, supplementation is not an effective strategy for the primary prevention of NTDs because of poor compliance. Thus, food fortification is seen by many as the only option likely to succeed. Mandatory folic acid fortification of grain products was introduced recently in the United States at a level projected to provide an additional mean intake of 100 microg folic acid/d, but some feel that this policy does not go far enough. A recent clinical trial predicted that the additional intake of folic acid in the United States will reduce NTDs by >20%, whereas 200 microg/d would be highly protective and is the dose also shown to be optimal in lowering plasma homocysteine, with possible benefits in preventing cardiovascular disease. Thus, an amount lower than the current target of an extra 400 microg/d may be sufficient to increase red blood cell folate to concentrations associated with the lowest risk of NTDs, but further investigation is warranted to establish the optimal amount. PMID- 10799408 TI - Maternal calcium metabolism and bone mineral status. AB - Human pregnancy is associated with major changes in calcium and bone metabolism and in bone mineral status before and after gestation. The changes are compatible with the uptake and mobilization of calcium by the maternal skeleton to meet the high requirement for fetal growth and for breast-milk production. Breast-feeding is accompanied by decreases in bone mineral status, increases in bone turnover rate, and reductions in urinary calcium excretion. These effects are reversed during and after weaning, and, in several skeletal regions, bone mineral content ultimately exceeds that measured after delivery. By 3-6 mo after lactation, the postpartum changes in bone mineral status of women who breast-feed largely match those of women who do not, regardless of the duration of lactation. No consistent picture has emerged of the effect of pregnancy on bone mineral status, although increases in bone turnover, calcium absorption, and urinary calcium excretion are well recognized. Events before conception may modify the bone response, particularly if conception occurs within a few months of a previous pregnancy or lactation. There is no evidence that the changes observed during lactation reflect inadequacies in calcium intake. Supplementation studies have shown that neither the bone response nor breast-milk calcium secretion is modified by increases in calcium supply during lactation, even in women with a low calcium intake. The situation in pregnancy is less clear. Calcium nutrition may influence the health of the pregnant woman, her breast-milk calcium concentration, and the bone mineralization and blood pressure of her infant, but these possibilities require formal testing. PMID- 10799409 TI - Perinatal metabolism of vitamin D. AB - During pregnancy, maternal serum concentrations of 25-hydroxyvitamin D, the circulating form of vitamin D, correlate with dietary vitamin D intake. Maternal serum concentrations of 1,25-dihydroxyvitamin D, the hormonal circulating and active form of vitamin D, are elevated during pregnancy; 1,25-dihydroxyvitamin D is synthesized mainly by the decidual cells of the placenta and allows for increased calcium absorption. The fetus is entirely dependent on the mother for its supply of 25-hydroxyvitamin D, which is believed to cross the placenta. Hypocalcemia and increased parathyroid hormone secretion induce synthesis of 1,25 dihydroxyvitamin D after birth in both full-term and preterm neonates. Nevertheless, serum concentrations of 25-hydroxyvitamin D are a rate-limiting factor in the synthesis of 1,25-dihydroxyvitamin D. In vitamin D-replete infants, circulating 1,25-dihydroxyvitamin D concentrations are higher than those observed in older infants. In countries where dairy products are not routinely supplemented with vitamin D, maternal vitamin D supplementation during pregnancy is necessary. However, there is no indication for the use of pharmacologic doses of vitamin D or its metabolites in the perinatal period. PMID- 10799410 TI - Vitamin A in pregnancy: requirements and safety limits. AB - Most of the functions of vitamin A are mediated through the binding of retinoic acid to specific nuclear receptors that regulate genomic expression. Recent experimental work in transgenic mice showed clearly that normal embryonic development depends on the correct spatial and temporal expression of the receptors in the differentiating cells and on the binding of specific forms of retinoic acid. This implies that the parent compound, vitamin A, is available in adequate forms and quantities. Excessive dietary intake of vitamin A has been associated with teratogenicity in humans in <20 reported cases over 30 y. However, caution must be exercised to avoid unnecessary supplementation of women of childbearing age. Hypovitaminosis A affects millions of women and children worldwide. The main consequence of a poor vitamin A supply during pregnancy is a low vitamin A status at birth and in the next few months. Vitamin A deficiency is strongly associated with depressed immune function and higher morbidity and mortality due to infectious diseases such as diarrhea, measles, and respiratory infections. Vitamin A deficiency is often associated with an increased mother-to child transmission of HIV-1. The initiation of vitamin A supplementation should be carefully examined in each case according to the risk-to-benefit ratio. The final decision should take into account the estimated vitamin A status of the woman, the availability of vitamin A-rich foods in her diet, and whether supplementation can be supervised. PMID- 10799411 TI - Determinants of maternal zinc status during pregnancy. AB - Zinc deficiency in pregnant experimental animals limits fetal growth and, if severe, causes teratogenic anomalies. Although the data from human studies are not consistent, similar outcomes have been observed and were associated with poor maternal zinc status. This paper reviews humans studies of zinc status and pregnancy outcome, describes the physiologic adjustments in zinc utilization during pregnancy to meet fetal needs while maintaining maternal status, and identifies dietary and environmental conditions that may override those physiologic adjustments and put the health of the mother and fetus at risk. Adjustments in intestinal zinc absorption appear to be the primary means by which zinc retention is increased to meet fetal demands. However, transfer of sufficient zinc to the fetus is dependent on maintenance of normal maternal serum zinc concentrations. Conditions that could interfere with zinc absorption include intake of cereal-based diets that are high in phytate, high intakes of supplemental iron, or any gastrointestinal disease. Conditions that may alter maternal plasma zinc concentrations and the transport of zinc to the fetus include smoking, alcohol abuse, and an acute stress response to infection or trauma. Supplemental zinc may be prudent for women with poor gastrointestinal function or with any of these conditions during pregnancy. PMID- 10799412 TI - Fetal nutrition and adult disease. AB - Recent research suggests that several of the major diseases of later life, including coronary heart disease, hypertension, and type 2 diabetes, originate in impaired intrauterine growth and development. These diseases may be consequences of "programming," whereby a stimulus or insult at a critical, sensitive period of early life has permanent effects on structure, physiology, and metabolism. Evidence that coronary heart disease, hypertension, and diabetes are programmed came from longitudinal studies of 25,000 UK men and women in which size at birth was related to the occurrence of the disease in middle age. People who were small or disproportionate (thin or short) at birth had high rates of coronary heart disease, high blood pressure, high cholesterol concentrations, and abnormal glucose-insulin metabolism. These relations were independent of the length of gestation, suggesting that cardiovascular disease is linked to fetal growth restriction rather than to premature birth. Replication of the UK findings has led to wide acceptance that low rates of fetal growth are associated with cardiovascular disease in later life. Impaired growth and development in utero seem to be widespread in the population, affecting many babies whose birth weights are within the normal range. Although the influences that impair fetal development and program adult cardiovascular disease remain to be defined, there are strong pointers to the importance of the fetal adaptations invoked when the maternoplacental nutrient supply fails to match the fetal nutrient demand. PMID- 10799413 TI - Improving prenatal nutrition in developing countries: strategies, prospects, and challenges. AB - In developing countries, the health and nutrition of females throughout their entire life is affected by complex and highly interrelated biological, social, cultural, and health service-related factors. Rather than focusing exclusively on the prenatal period, we describe a life cycle approach to improving maternal nutrition, which goes beyond the traditional provision of nutrition services during pregnancy, by addressing risk factors that are present well before pregnancy, even before childbearing age. This approach involves specific policy initiatives and a "minimum package" program that is targeted at females. Policy actions and the components for effective implementation of the program are described. The prospects and challenges to be overcome-which include translating scientific knowledge into action, removing conceptual and implementational constraints, identifying biologically meaningful indicators for problem identification, and improving understanding of physiologic and social adaptation mechanisms-are discussed, as are persistent problems with health care delivery systems. PMID- 10799414 TI - Sociocultural and behavioral influences on weight gain during pregnancy. AB - Studies have consistently identified a positive association between prenatal weight gain and birth weight. Much less, however, is known about factors that may influence women to gain weight within currently recommended ranges. The importance of this issue is suggested by recent reports indicating that only 30 40% of women actually gain weight within these ranges. This paper examines demographic, sociocultural, and behavioral factors that are associated with, and may influence risk of, low prenatal weight gain among adult women with low and normal body mass indexes. Available data suggest that these factors include ethnicity, socioeconomic status, age, education, pregnancy intendedness or wantedness, prenatal advice, and psychosocial characteristics such as attitude toward weight gain, social support, depression, stress, anxiety, and self efficacy. Potential theoretical models for these associations include biological, behavioral, and mixed pathways. The design of targeted intervention studies will depend on further identification and characterization of sociocultural and behavioral risk factors that, along with reproductive and nutritional characteristics, may predict which women are most likely to have inadequate prenatal weight gain. PMID- 10799415 TI - Dietary calcium and pregnancy-induced hypertension: is there a relation? AB - The evidence that calcium plays a role in the etiology, prevention, and treatment of pregnancy-induced hypertension (PIH) is reviewed. The precise factors involved in the pathogenesis of PIH are unclear, but several alterations in calcium metabolism have been identified. Epidemiologic data suggest an inverse correlation between dietary calcium intake and incidence of PIH. Although evidence suggests a possible beneficial effect of supplemental calcium, contradictions persist in clinical trials of pregnant women. Presently, there is insufficient evidence to support routine calcium supplementation of all pregnant women. However, high-risk groups, such as pregnant teens, populations with inadequate calcium intake, and women at risk of developing PIH, may benefit from consuming additional dietary calcium. PMID- 10799416 TI - Same nutrient, different hypotheses: disparities in trials of calcium supplementation during pregnancy. AB - Calcium supplementation during pregnancy has been provided either to increase the intake in those with a deficiency or to obtain a pharmacologic, perhaps nonnutritional, effect in individuals with an adequate calcium intake. A systematic review, including only randomized, double-blind, controlled trials of calcium supplementation during pregnancy was prepared independently for the Cochrane Library and updated by us for this paper. In view of the heterogeneity of results included in the meta-analysis, a stratified analysis by baseline dietary calcium intake (mean calcium intake in the population < or >/=900 mg/d) was conducted. On the basis of the results of the 5 randomized, controlled trials available, the risk of high blood pressure was lower in women with low baseline dietary calcium [typical relative risk (TRR): 0.49; 95% CI: 0.38, 0.62]. Of the 4 trials in which subjects had adequate dietary calcium, the TRR of high blood pressure was 0.90 (95% CI: 0.81, 0.99). The risk of preeclampsia was considerably reduced in the 6 trials conducted in populations with low-calcium diets (TRR: 0.32; 95% CI: 0.21, 0.49) but was not reduced as much in women enrolled in the 4 trials with adequate-calcium diets (TRR: 0.86; 95% CI: 0.71, 1.05). On the basis of these results, it seems clear that calcium supplementation during pregnancy for women with deficient calcium intake is a promising preventive strategy for preeclampsia. Calcium supplementation in pregnancy should be evaluated definitively in an adequately sized trial conducted in a population with a low calcium intake because this is the most likely population to benefit from such a nutritional intervention. Long-term health benefits for the offspring are also an attractive possibility. PMID- 10799417 TI - The biology of mitochondrial disease. PMID- 10799418 TI - Stamps in paediatrics. Infectious diseases. PMID- 10799419 TI - Systematic review of controlled trials of interventions to promote smoke alarms. AB - AIMS: To evaluate the effects of promotion of residential smoke alarms. METHODS: Electronic databases, conference proceedings, and bibliographies were systematically searched, and investigators and organisations were contacted, in order to identify controlled trials evaluating interventions designed to promote residential smoke alarms. The following were assessed: smoke alarm acquisition, ownership, and function; fires; burns; and fire related injuries. Odds ratios (OR) were estimated by meta analysis of randomised trials. RESULTS: A total of 26 trials were identified, of which 13 were randomised. Overall, counselling and educational interventions had only a modest effect on the likelihood of owning an alarm (OR = 1.26; 95% confidence interval (CI): 0.87 to 1.81) or having a functional alarm (OR = 1.19; 95% CI: 0.85 to 1.66). Counselling as part of primary care child health surveillance had greater effects on ownership (OR = 1.93; 95% CI: 1.04 to 3.58) and function (OR = 1. 72; 95% CI: 0.78 to 3.78). Results were sensitive to trial quality, however, and effects on fire related injuries were not reported. In two non-randomised trials, direct provision of free alarms significantly increased functioning alarms and reduced fire related injuries. Media and community education showed little benefit in non-randomised trials. CONCLUSION: Counselling as part of child health surveillance may increase smoke alarm ownership and function, but its effects on injuries are unevaluated. Community smoke alarm give away programmes apparently reduce fire related injuries, but these trials were not randomised and results must be interpreted cautiously. Further efforts to promote smoke alarms in primary care or through give away programmes should be evaluated by adequately designed randomised controlled trials measuring injury outcomes. PMID- 10799420 TI - Socioeconomic inequalities in risk of congenital anomaly. AB - AIMS: To investigate socioeconomic inequalities in the risk of congenital anomalies, focusing on risk of specific anomaly subgroups. METHODS: A total of 858 cases of congenital anomaly and 1764 non-malformed control births were collected between 1986 and 1993 from four UK congenital malformation registers, for the purposes of a European multicentre case control study on congenital anomaly risk near hazardous waste landfill sites. As a measure of socioeconomic status, cases and controls were given a value for the area level Carstairs deprivation index, by linking the postcode of residence at birth to census enumeration districts (areas of approximately 150 households). RESULTS: Risk of non-chromosomal anomalies increased with increasing socioeconomic deprivation. The risk in the most deprived quintile of the deprivation index was 40% higher than in the most affluent quintile. Some malformation subgroups also showed increasing risk with increasing deprivation: all cardiac defects, malformations of the cardiac septa, malformations of the digestive system, and multiple malformations. No evidence for socioeconomic variation was found for other non chromosomal malformation groups, including neural tube defects and oral clefts. A decreasing risk with increasing deprivation found for all chromosomal malformations and Down's syndrome in unadjusted analyses, occurred mainly as a result of differences in the maternal age distribution between social classes. CONCLUSION: Our data, although based on limited numbers of cases and geographical coverage, suggest that more deprived populations have a higher risk of congenital anomalies of non-chromosomal origin and some specific anomalies. Larger studies are needed to confirm these findings and to explore their aetiological implications. PMID- 10799421 TI - Academic career after treatment for acute lymphoblastic leukaemia. AB - AIM: To evaluate academic career in long term survivors of childhood acute lymphoblastic leukaemia (ALL), in comparison to their healthy siblings. PATIENTS: Ninety four children treated for ALL with cranial irradiation 18 or 25 Gy and intrathecal methotrexate as CNS prophylaxis. Median age at evaluation was 20 years; median follow up since diagnosis was 15 years at the time of the study. METHODS: Patients and their 134 siblings completed a questionnaire on school career. The percentage of referrals to special primary schools for learning disabled, and the final level of secondary education in patients and siblings were compared, using a six point classification. Within the patient group, the effect of possible risk factors (age at diagnosis, irradiation dose, and gender) was investigated. RESULTS: Significantly more patients than siblings were placed in special educational programmes. A significant difference was found for level of secondary education. No effect of gender or irradiation dose was found, but younger age at diagnosis was significantly related to both referrals and school levels. CONCLUSION: Treatment for childhood ALL with cranial irradiation and chemotherapy at a young age is clearly associated with poorer academic career. PMID- 10799422 TI - Circumcision in the United States-the debate continues. PMID- 10799424 TI - Anthropometric indices of failure to thrive. AB - AIMS: To compare five anthropometric methods of classifying failure to thrive in order to ascertain their relative merits in predicting developmental, dietary, and eating problems. METHODS: The five anthropometric methods were compared in 83 children with failure to thrive. RESULTS: The methods were inconsistent in classification of severity, and no one method was superior in predicting problems. CONCLUSIONS: Weight alone, being the simplest, is still the most reasonable marker for failure to thrive and associated problems. PMID- 10799423 TI - Prepubertal stature and blood pressure in early old age. AB - AIMS: To test the hypothesis that childhood growth rate is a marker for formation of control mechanisms that influence blood pressure in early old age. METHODS: Data are from a sample of 149 (74 male) members of Sir John Boyd Orr's survey of British families conducted between 1937 and 1939. Measured heights were collected between ages 5 and 8 years, and in early old age between 1997 and 1998. Multiple linear regression investigated the relations of blood pressure with age and sex standardised childhood height with adjustment for potential confounding factors, including adult height. Inclusion of both childhood and adult heights in the same model was used to estimate growth, as measures of childhood height are relative to adult height. RESULTS: Mean blood pressures in early old age for those in the shortest childhood height fifth were 167.8 and 76.3 mm Hg for systolic blood pressure and pulse pressure, respectively. For the tallest fifth they were 150.8 and 63.7 mm Hg, respectively. After adjustment for potential confounding factors including adult height, the mean increase for the shortest childhood height fifth compared with the tallest was 28.5 mm Hg for systolic pressure (p = 0.015) and 22.8 mm Hg (p = 0.010) for pulse pressure. The relations of blood pressure with adult height were not statistically significant in the adjusted models. CONCLUSION: Prepubertal growth rate is associated with the formation of mechanisms associated with the control of blood pressure in later life. PMID- 10799425 TI - Fish oil supplementation improves docosahexaenoic acid status of malnourished infants. AB - AIM: To investigate whether the low docosahexaenoic acid (DHA) status of malnourished, mostly breast fed, Pakistani children can be improved by fish oil (FO) supplementation. METHODS: Ten malnourished children (aged 8-30 months) received 500 mg FO daily for nine weeks. The supplement contained 62.8 mol% (314 mg) long chain polyunsaturated fatty acids of the omega3 series (LCPUFAomega3) and 22.5 mol% (112 mg) DHA. Seven FO unsupplemented children served as controls. Red blood cell (RBC) fatty acids were analysed at baseline and at the study end. RESULTS: FO supplementation augmented mean (SD) RBC DHA from 2.27 (0.81) to 3.35 (0.76) mol%, without significantly affecting the concentrations of LCPUFAomega6. Unsupplemented children showed no RBC fatty acid changes. One FO supplemented child with very low initial RBC arachidonic acid showed a remarkable increase from 4.04 to 13.84 mol%, whereas another with high RBC arachidonic acid showed a decrease from 15.64 to 10.46 mol%. CONCLUSION: FO supplementation improves the DHA status of malnourished children. The supplement is apparently well absorbed and not exclusively used as a source of energy. PMID- 10799426 TI - Inhaled salbutamol for wheezy infants: a randomised controlled trial. AB - BACKGROUND: Salbutamol is frequently used as a bronchodilator for infants who wheeze. Many single dose studies have questioned its effectiveness. AIMS: To investigate the response of wheezy infants to salbutamol over an extended time period in order to elucidate either symptomatic relief or a protective effect. METHODS: Eighty infants under 1 year, with persistent or recurrent wheeze and a personal or family history of atopy, were recruited to a randomised, double blind, cross over, placebo controlled trial. Salbutamol (200 microg three times daily) or placebo were administered regularly over two consecutive treatment periods of four weeks via a spacer and mask. Symptoms of wheeze and cough were recorded in a diary. At the end of the study pulmonary function tests were performed before and after salbutamol (400 microg). RESULTS: Forty eight infants completed the diary study; 40 infants underwent pulmonary function testing. No difference in mean daily symptom score was observed between the salbutamol and placebo periods. There was no difference in the number of symptom free days. Compliance and forced expiratory flows remained unchanged and resistance increased following salbutamol. There was no relation between the response measured by symptom score or pulmonary function in individual patients. CONCLUSION: In wheezy infants with an atopic background, there was no significant beneficial effect of salbutamol on either clinical symptoms or pulmonary function. Clinical effects could not be predicted from pulmonary function tests. Salbutamol cannot be recommended as the bronchodilator of choice in this age group. PMID- 10799427 TI - Risk assessment of renal cortical scarring with urinary tract infection by clinical features and ultrasonography. AB - AIMS: To address some of the issues in the ongoing debate over the optimal diagnostic imaging following childhood urinary tract infection (UTI), by determining the risk of missing renal cortical scarring which would be detected on a technetium-99m dimercaptosuccinic acid (DMSA) gold standard if ultrasound alone were used, factoring for clinical features (upper or lower tract), UTI recurrence, and age group (infants, preschool, or school age). METHODS: Details of UTI clinical features and recurrence were recorded for 990 children with a proven UTI, and their DMSA and ultrasound results were compared for each kidney. RESULTS: The risks of missing DMSA scarring varied between 0.4% (school age children with solitary lower tract UTI) and 11.1% (infants with recurrent upper tract UTI). CONCLUSIONS: UTI clinical features are important in assessing the need for DMSA imaging. Current UK imaging guidelines are endorsed, although preschool children with solitary lower tract UTI remain a controversial group and more attention needs to focused on children with recurrent UTI. PMID- 10799428 TI - Visceral leishmaniasis in Malta--an 18 year paediatric, population based study. AB - BACKGROUND: Visceral leishmaniasis (VL) is a chronic parasitic infection that infects approximately 400,000 individuals annually, with a predilection towards early childhood. AIMS: To study the epidemiology of VL in childhood. METHODS: VL is endemic in Malta, a small archipelago of islands in the centre of the Mediterranean with a total population approaching half a million. Notification of human cases of leishmaniasis is compulsory. Case records of all 81 paediatric patients with VL between 1980 and 1998 were analysed. RESULTS: The annual incidence of VL declined for all cases of VL, and declined significantly for paediatric cases (p = 0.01). For 1994 to 1998, the overall incidence of VL was 0.9 per 100,000 total population and the paediatric incidence was 2.5 per 100,000 population. Median age at presentation was 34 months. Common features at presentation were splenomegaly, hepatomegaly, fever, and pancytopenia with high lymphocyte and monocyte counts. The diagnostic sensitivity of isolated immunofluorescent antibody testing was equivalent to bone marrow aspiration (95%). Blood transfusions for anaemia were required in 93% of patients. Eleven per cent had intercurrent infections. All patients were cured, and were initially treated with intravenous sodium stibogluconate. Defervescence occurred after a median of six days of treatment, and patients continued to be treated on a day case basis. Nine relapsers were retreated with sodium stibogluconate, achieving a cure rate of 94%, but five patients required additional drug therapy. There were no permanent sequelae associated with VL or its treatment. CONCLUSIONS: The decreased incidence is attributed to the eradication of stray dogs which are the disease reservoir. PMID- 10799429 TI - Epidemiology of Perthes' disease. PMID- 10799430 TI - High incidence of primary tuberculosis. AB - An usually high rate of both tuberculosis infection and active disease is reported in 11 of 38 nursery children in contact with a case of smear positive pulmonary tuberculosis, emphasising the susceptibility of young children to this disease. This report also underlines some important principles in case finding and disease control. PMID- 10799431 TI - Angiotensin for septic shock unresponsive to noradrenaline. AB - Two children with severe septic shock are reported. One had meningococcal septicaemia and the other Escherichia coli septicaemia. They remained hypotensive despite high concentrations of conventional inotropes and vasopressors. In one child, using a pulmonary artery catheter, extended haemodynamic variables were measured. To restore blood pressure, in both cases, an infusion of angiotensin II was used; there was significant improvement in clinical status, resulting in a rapid reduction in the concentration of inotropes required. Both patients successfully survived their septic episodes. Angiotensin II in cases of severe refractory septic hypotension in the paediatric population offers an extra therapeutic manoeuvre. PMID- 10799432 TI - Under recognition of late onset ornithine transcarbamylase deficiency. AB - Late onset ornithine transcarbamylase deficiency (McKusick 311250) is reported in four Finnish patients, two boys and two heterozygous girls. The subtle onset and course of ornithine transcarbamylase deficiency emphasises the need for plasma ammonia and amino acid measurements in clinical situations suggesting a disorder of this nature. PMID- 10799434 TI - FETAL AND NEONATAL EDITION may 2000 issue PMID- 10799433 TI - Deliberate sulphonylurea poisoning mimicking hyperinsulinaemia of infancy. AB - A 6 month old child presenting with seizures was found to be hypoglycaemic secondary to hyperinsulinism. A family history of type II diabetes prompted estimation of sulphonylurea in the baby's blood, which was found to be high. A multidisciplinary case conference concluded that the sulphonylurea ingestion was likely to be the result of Munchausen syndrome by proxy. When investigating hypoglycaemia of infancy this possibility should be considered. PMID- 10799435 TI - Intestinal inflammation in cystic fibrosis. AB - BACKGROUND: There is controversy about whether the inflammatory response observed in the cystic fibrosis (CF) lung occurs secondary to bacterial infection or is caused by a dysregulation of the inflammatory response associated with the basic cellular defect of CF. AIMS: To study the inflammatory response in the gastrointestinal tract of children with CF; and to investigate whether there is increased inflammation in the gastrointestinal tract of CF children with fibrosing colonopathy. METHODS: Whole gut lavage was performed on 21 pancreatic insufficient children with CF, who were clinically well, five children with CF and fibrosing colonopathy, and 12 controls. Intestinal outputs of plasma derived proteins (albumin, alpha(1) antitrypsin, IgG), secretory immunoglobulins (IgA and IgM), cellular constituents (eosinophil cationic protein and neutrophil elastase), and cytokines (interleukin 8 and interleukin 1beta) were measured. RESULTS: Compared to controls, the 21 CF patients, with no intestinal complications, had increased intestinal outputs of albumin, IgG, IgM, eosinophil cationic protein, neutrophil elastase, interleukin 1beta, and interleukin 8. Similar values were obtained for the CF patients with fibrosing colonopathy. CONCLUSIONS: These data suggest that there is immune activation in the gastrointestinal mucosa of children with cystic fibrosis, which may result from the basic cellular defect. Fibrosing colonopathy does not appear to be associated with increased inflammation. PMID- 10799436 TI - Nijmegen breakage syndrome. The International Nijmegen Breakage Syndrome Study Group. AB - BACKGROUND: Nijmegen breakage syndrome (NBS) is a rare autosomal recessive disorder. NBS-1, the gene defective in NBS, is located on chromosome 8q21 and has recently been cloned. The gene product, nibrin, is a novel protein, which is member of the hMre11/hRad50 protein complex, suggesting that the gene is involved in DNA double strand break repair. AIMS: To study the clinical and laboratory features of NBS as well as the genotype-phenotype relation. METHODS: Fifty five patients with NBS, included in the NBS registry in Nijmegen were evaluated. The majority of the patients were of eastern European ancestry. Most of them had shown a truncating 5 bp deletion 657-661 delACAAA. Four further truncating mutations have been identified in patients with other distinct haplotypes. RESULTS AND CONCLUSIONS: Essential features found in NBS were microcephaly, usually without severe retardation, typical facial appearance, immunodeficiency, chromosomal instability, x ray hypersensitivity, and predisposition to malignancy. In 40% of the patients cancer was noted before the age of 21 years. Important additional features were skin abnormalities, particularly cafe au lait spots and vitiligo, and congenital malformations, particularly clinodactyly and syndactyly. Congenital malformations, immunodeficiency, radiation hypersensitivity, and cancer predisposition were comprehensible in case of dysfunctioning of DNA repair mechanisms. No specific genotype-phenotype relation could be found. Patients with the same genotype may show different phenotypes and patients with different genotypes may express the same phenotype. Specific mutations did not lead to specific clinical features. PMID- 10799437 TI - Heterogeneous presentation in A3243G mutation in the mitochondrial tRNA(Leu(UUR)) gene. AB - AIMS: To clarify the phenotype-genotype relation associated with the A3243G mitochondrial DNA mutation. METHODS: Five unrelated probands harbouring the A3243G mutation but presenting different clinical phenotype were analysed. Probands include Leigh syndrome (LS(3243)), mitochondrial myopathy, encephalopathy, lactic acidosis and stroke like episodes (MELAS(3243)), progressive external ophthalmoplegia (PEO(3243)), and mitochondrial diabetes mellitus (MDM(3243)). Extensive clinical, histological, biochemical, and molecular genetic studies were performed on five families. RESULTS: All patients showed ragged red fibres (RRF), and focal cytochrome c oxidase (COX) deficiency except for the patient with MDM(3243). The mutation load was highest in the proband with LS(3243) (>90%), who also presented the highest proportion of RRF (68%) and COX negative fibres (10%), and severe complex I plus IV deficiency. These proportions were lower in the probands with PEO(3243) and with MDM(3243). CONCLUSION: The most severe clinical phenotype, LS(3243), was associated with the highest proportion of the A3243G mutation as well as the most prominent histological and biochemical abnormalities. PMID- 10799438 TI - Neuroimaging in child and adolescent psychiatric disorders. AB - Neuroimaging in child psychiatry is a rapidly developing field and the number of different techniques being used is increasing rapidly. This review describes the current status of neuroimaging in childhood psychopathology and discusses limitations of the various studies. As yet, no specific and consistent abnormality has been detected in childhood psychiatric disorders. Obsessive compulsive disorder has shown the most consistent findings so far, with orbitofrontal cortex and the caudate nucleus being implicated. Better understanding of the corticostriatal neural networks will shed more light on the neurodevelopmental disorders of childhood. PMID- 10799440 TI - Inconsistencies in sweat testing in UK laboratories. AB - BACKGROUND: Sweat testing procedures are perceived to vary widely. AIM: To evaluate variability in sweat collection, analysis, and interpretation. METHODS: Questionnaire responses from 30 self selected centres: 15 paediatric centres, and 15 district general hospitals. RESULTS: Centres carried out 30-400 tests per year (median 100), with a diagnostic rate of 1:5-152 (median 1:30). Staff performed 5 268 tests per year. Minimum test age varied from 24 hours to four months. All stimulated sweating by pilocarpine iontophoresis using varying currents and times. Twenty six had observed urticaria or skin reddening, and nine blistering or burns. Sweat was collected for 10-60 minutes onto filter paper or into Macroduct coils. Between 2% and 25% of tests were considered insufficient. Twenty eight measured sodium, 24 chloride, and one osmolality and conductivity. Fifteen used literature and five in house reference ranges. Eleven would not test severely eczematous children. CONCLUSIONS: Local audit is required to improve performance, as well as a national guideline to standardise collection, and external quality assessment to provide analytical feedback. PMID- 10799439 TI - Indirect measurements of sweat electrolyte concentration in the laboratory diagnosis of cystic fibrosis. AB - AIM: To investigate whether analytical methods based on the colligative physical chemical properties of ions or solutes in sweat are less effective than the specific measurement of electrolytes in the diagnosis of cystic fibrosis (CF). METHODS: A single sweat sample was collected (Macroduct) from each of 211 infants and children, of whom 57 had CF, for the measurement of sodium, chloride, osmolality, and conductivity. RESULTS: The ranges within which CF and non-CF individual values overlapped (equivocal ranges), were wider for sodium and osmolality measurement than for chloride or conductivity. Neither of the latter two measurements provided a discriminatory advantage over the other. The utilisation of broadly based age related ranges for non-CF control subjects served to improve the discriminatory power of all four measurements to an extent that, in this cohort, both chloride and conductivity provided complete discrimination. CONCLUSION: Sweat conductivity is as effective as chloride measurement in the laboratory diagnosis of CF. PMID- 10799441 TI - Monitoring of antigen-specific cytolytic T lymphocytes in cancer patients receiving immunotherapy. PMID- 10799442 TI - Adoption of single-platform technologies for enumeration of absolute T-lymphocyte subsets in peripheral blood. PMID- 10799443 TI - Evaluation of TruCount absolute-count tubes for determining CD4 and CD8 cell numbers in human immunodeficiency virus-positive adults. Site Investigators and The NIAID DAIDS New Technologies Evaluation Group. AB - A single-platform technology that uses an internal bead standard and three-color flow cytometry to determine CD4 and CD8 absolute counts was evaluated for reproducibility and agreement. Values obtained using TruCount absolute-count tubes were compared to those obtained using a two-color predicate methodology. Sixty specimens from human immunodeficiency virus type 1-infected donors were shipped to five laboratories. Each site also analyzed replicates of 14 human immunodeficiency virus type 1-infected local specimens at 6 h and again at 24 h. The interlaboratory variability was significantly less with TruCount (median difference in percent coefficient of variation [%CV] between the two methods was 8% and -3% for CD4 and CD8, respectively) than with the predicate method. Intralaboratory variability was smaller, with a median difference in %CV of -1% for both CD4 and CD8 with 6-h samples and -2% and -3% for CD4 and CD8, respectively, with 24-h samples. Use of TruCount for shipped samples resulted in a median CD4 count change of 7 cells (50th estimated percentile) when all laboratories and CD4 strata were combined. For on-site samples, the median CD4 count change was 10 CD4 cells for 6-h samples and 2 CD4 cells for 24-h samples. Individual site biases occurred in both directions and cancelled each other when the data were combined for all laboratories. Thus, the combined data showed a smaller change in median CD4 count than what may have occurred at an individual site. In summary, the use of TruCount decreased both the inter- and intralaboratory variability in determining absolute CD4 and CD8 counts. PMID- 10799444 TI - Multisite comparison of CD4 and CD8 T-lymphocyte counting by single- versus multiple-platform methodologies: evaluation of Beckman Coulter flow-count fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group. AB - New analytic methods that permit absolute CD4 and CD8 T-cell determinations to be performed entirely on the flow cytometer have the potential for improving assay precision and accuracy. In a multisite trial, we compared two different single platform assay methods with a predicate two-color assay in which the absolute lymphocyte count was derived by conventional hematology. A two-color method employing lymphocyte light scatter gating and Beckman Coulter Flow-Count fluorospheres for absolute counting produced within-laboratory precision equivalent to that of the two-color predicate method, as measured by coefficient of variation of replicate measurements. The fully automated Beckman Coulter tetraONE System four-color assay employing CD45 lymphocyte gating, automated analysis, and absolute counting by fluorospheres resulted in a small but significant improvement in the within-laboratory precision of CD4 and CD8 cell counts and percentages suggesting that the CD45 lymphocyte gating and automated analysis might have contributed to the improved performance. Both the two-color method employing Flow-Count fluorospheres and the four-color tetraONE System provided significant and substantial improvements in between-laboratory precision of absolute counts. In some laboratories, absolute counts obtained by the single platform methods showed small but consistent differences relative to the predicate method. Comparison of each laboratory's absolute counts with the five laboratory median value suggested that these differences resulted from a bias in the absolute lymphocyte count obtained from the hematology instrument in some laboratories. These results demonstrate the potential for single-platform assay methods to improve within-laboratory and between-laboratory precision of CD4 and CD8 T-cell determinations compared with conventional assay methods. PMID- 10799446 TI - Relevance of the viral RAK alpha gene in diagnosis of malignant versus nonmalignant tumors of the ovary and uterus. AB - Human immunodeficiency virus type 1 (HIV-1)-like antigens RAK (named after the inventor E. M. Rakowicz) p120, p42, and p25, as well as HIV-1-like segments of cancer DNA (RAK gene alpha), have been found before in breast and prostate cancers. The present study focused on determining the value of markers RAK in the diagnosis and prognosis of gynecological cancer. Expression of RAK antigens in ovarian, uterine, cervical, and vulvar cancer, in benign tumors, in tissues adjacent to cancer, and in normal tissues was tested by Western blot hybridization of the electrophoretically separated proteins with monoclonal antibody RAK BrI. The RAK alpha gene was PCR amplified with HIV-1-derived primers SK68 and SK69. RAK antigens p120, p42, and p25 were found in 95% of ovarian, uterine, and cervical cancer cases and in 75% of vulvar cancer cases. The RAK alpha gene was expressed in 100% of cancer cases, in approximately 25% of benign ovarian tumors, and in 40% of benign tumors of the uterus. DNA sequences amplified in all cancer cases exhibited more than 90% homology to HIV-1 gp41 and were encoded for the functional peptide. DNA sequences found in benign tumors contained frameshift mutations and encoded truncated or nonfunctional peptides. Such sequences have not been amplified in normal tissues. RAK antigens and the RAK alpha gene seem to belong to a lentivirus type that is highly related to HIV 1. Beyond the diagnostic value of RAK markers, future cloning of the full viral genome would lead to a better understanding of the etiology of malignant and nonmalignant tumors of reproductive organs and to the development of novel therapeutic approaches. PMID- 10799445 TI - Preservation of lymphocyte immunophenotype and proliferative responses in cryopreserved peripheral blood mononuclear cells from human immunodeficiency virus type 1-infected donors: implications for multicenter clinical trials. The ACTG Immunology Advanced Technology Laboratories. AB - Human immunodeficiency virus type 1 (HIV-1) infection results in impaired immune function that can be measured by changes in immunophenotypically defined lymphocyte subsets and other in vitro functional assays. These in vitro assays may also serve as early indicators of efficacy when new therapeutic strategies for HIV-1 infection are being evaluated. However, the use of in vitro assays of immune function in multicenter clinical trials has been hindered by their need to be performed on fresh specimens. We assessed the feasibility of using cryopreserved peripheral blood mononuclear cells (PBMC) for lymphocyte immunophenotyping and for lymphocyte proliferation at nine laboratories. In HIV-1 infected patients with moderate CD4(+) lymphocyte loss, the procedures of density gradient isolation, cryopreservation, and thawing of PBMC resulted in significant loss of CD19(+) B cells but no measurable loss of total T cells or CD4(+) or CD8(+) T cells. No significant changes were seen in CD28(-) CD95(+) lymphocytes after cell isolation and cryopreservation. However, small decreases in HLA-DR(+) CD38(+) lymphocytes and of CD45RA(+) CD62L(+) were observed within both the CD4(+) and CD8(+) subsets. Fewer than 10% of those specimens that showed positive PBMC proliferative responses to mitogens or microbial antigens lost their responsiveness after cryopreservation. These results support the feasibility of cryopreserving PBMC for immunophenotyping and functional testing in multicenter AIDS clinical trials. However, small changes in selected lymphocyte subsets that may occur after PBMC isolation and cryopreservation will need to be assessed and considered in the design of each clinical trial. PMID- 10799447 TI - Induction of hypergammaglobulinemia and macrophage activation by silicone gels and oils in female A.SW mice. AB - Although most published epidemiological studies have found little evidence of systemic autoimmune disease associated with silicone breast implants, there still remains a question of whether silicones can cause local and/or systemic immune dysfunction. This study further investigates the effects of silicones on autoantibody and immunoglobulin production and macrophage activation in female A.SW mice. Sixty mice were divided among four treatment groups receiving a 0.5-ml intraperitoneal injection of either phosphate-buffered saline (PBS), pristane, silicone gel, or silicone oil. Test bleeds were taken periodically for 6 months. In contrast to pristane, neither silicone gel nor silicone oil induced lupus associated antinuclear autoantibodies (immunoglobulin G [IgG] anti-nRNP/Sm, Su, and ribosomal P) or lupus nephritis. However, serum IgM became elevated persistently within 1 month of silicone gel or silicone oil administration. Also, the level of IgG3 was clearly elevated in silicone oil-treated mice. In contrast, IgG1, IgG2a, and IgG2b levels were not affected greatly by either silicone gel or oil. Furthermore, peritoneal macrophages from silicone- and pristane-treated mice produced higher levels of interleukin-1beta (IL-1beta) and IL-6 than those from PBS-treated mice after lipopolysaccharide stimulation. These results suggest that silicone gels and oils are capable of inducing hypergammaglobulinemia and activating macrophages in female A.SW mice. PMID- 10799448 TI - Differential expression of neurokinin-1 receptor by human mucosal and peripheral lymphoid cells. AB - Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-1 receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM). Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-1R was evident in LPMC but not in PBMC, PBL, monocytes, or MDM. However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM. This level of expression was found to represent one NK-1R mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC. These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation. PMID- 10799449 TI - Variability and immunogenicity of human immunodeficiency virus type 1 p24 gene quasispecies. AB - Despite the conserved nature of the human immunodeficiency virus type 1 (HIV-1) gag gene, multiple quasispecies of the p24 gene coexist in HIV-1-infected patients. We cloned and sequenced 31 p24 genes from four HIV-1-infected patients. The intrapatient homology between the p24 genes ranged from 97.1 to 99.1%, whereas the interpatient homology ranged from 91.5 to 93.8%, suggesting a host specific evolution. Synonymous and nonsynonymous nucleotide changes were evenly distributed in the p24 gene, with 27 and 28%, respectively, located within host human leukocyte antigen class I recognition sites. This would suggest only a minor influence from the host cytotoxic T-cell response on the evolution of the p24 gene. The importance of minor variations within p24 was analyzed by designing DNA-based immunogens from two distinct p24 quasispecies genes simultaneously derived from one patient. In plasmid-immunized H-2(b), H-2(d), and H-2(k) haplotype mice, a clear influence from the host major histocompatibility complex was noted on the immune responses, fully consistent with those noted when a recombinant p24 protein is used as the immunogen. The two p24 DNA immunogens did not differ in their immunogenicity, indicating that the limited genetic variability (<1%) had little influence on the immune responses. PMID- 10799450 TI - Molecular markers in acute and chronic phases of human toxoplasmosis: determination of immunoglobulin G avidity by Western blotting. AB - We characterized antigenic markers recognized by human serum samples from patients presenting with acute and chronic toxoplasmosis by the determination of immunoglobulin G (IgG) antibody avidity by a Western blot modified technique (avidity immunoblotting) that includes the dissociation of the antigen-antibody interaction with 6 or 8 M urea solutions. Human serum samples from 20 patients presenting with recent infection and from 20 patients with chronic infection were analyzed. It was observed that bands p16, p32, p38, p40, p43, p54, p60, p66, and p97 were more frequently recognized by low-avidity IgG in recent infection and by high-avidity IgG in chronic toxoplasmosis. From these antigenic bands, p38 can be characterized as an optimal antigenic marker of low avidity for recent forms of toxoplasmosis due to a significant decrease of their frequencies (from 80 to 0%) after treatment with 6 M urea solutions. The p30 antigen was not considered a good marker to distinguish acute from chronic infection since corresponding IgG antibodies were determined to have high avidity in both phases of the infection. Thus, the avidity immunoblotting assay proved to be a useful tool for determining antigenic markers of recent and chronic phases of Toxoplasma gondii infection. PMID- 10799451 TI - Sequence variation in the porA gene of a clone of Neisseria meningitidis during epidemic spread. AB - The ET-15 clone within the electrophoretic type (ET)-37 complex of Neisseria meningitidis was first detected in Canada in 1986 and has since been associated with outbreaks of meningococcal disease in many parts of the world. While the majority of the strains of the ET-37 complex are serosubtype P1.5,2, serosubtype determination of ET-15 strains may often be incomplete, with either only one or none of the two variable regions (VRs) of the serosubtype PorA outer membrane protein reacting with monoclonal antibodies. DNA sequence analysis of the porA gene from ET-15 strains with one or both unidentified serosubtype determinants was undertaken to identify the genetic basis of the lack of reaction with the monoclonal antibodies. Fourteen different porA alleles were identified among 38 ET-15 strains from various geographic origins. The sequences corresponding to subtypes P1.5a,10d, P1.5,2, P1.5,10d, P1.5a,10k, and P1.5a,10a were identified in 18, 11, 2, 2, and 1 isolate, respectively. Of the remaining four strains, which all were nonserosubtypeable, two had a stop codon within the VR1 and the VR2, respectively, while in the other two the porA gene was interrupted by the insertion element, IS1301. Of the strains with P1.5,2 sequence, one had a stop codon between the VR1 and VR2, one had a four-amino-acid deletion outside the VR2, and another showed no expression of PorA on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Our results reveal that numerous genetic events have occurred in the porA gene of the ET-15 clone in the short time of its epidemic spread. The magnitude of microevolutionary mechanisms available in meningococci and the remarkable genetic flexibility of these bacteria need to be considered in relation to PorA vaccine development. PMID- 10799452 TI - Use of monoclonal antibodies for rapid diagnosis of respiratory viruses in a community hospital. AB - An indirect fluorescence antibody (IFA) procedure was used for the rapid detection of respiratory viruses in direct clinical specimens and for determining the epidemiology of viruses in a community hospital setting. Viral respiratory diseases were monitored for 10 consecutive respiratory seasons. The Bartels Viral Respiratory Screening and Identification Kit is an IFA method that contains pooled and individual monoclonal antibodies for seven common respiratory viruses. Compared with 8,670 conventional tube cell cultures, IFA staining of direct patient specimens had an overall sensitivity of 84.2% and a specificity of 87.7%. Yearly epidemics of respiratory syncytial virus were seen with alternating short and long intervals between successive periods when virus was isolated. Epidemics following short intervals were more severe. Influenza A virus epidemics occurred yearly, and influenza B virus activity was seen generally every other year. When influenza A and influenza B viruses were cocirculating in a given season, the months of peak activity of one virus were always within 1 month of the peak activity of the other virus. Parainfluenza virus type 1 was detected in the autumn of odd-numbered years, and parainfluenza type 2 virus was seen usually in the autumn of even-numbered years. Parainfluenza type 3 virus and adenovirus were the most ubiquitous agents, with peak incidence occurring in the late winter to spring. PMID- 10799453 TI - Development of an indirect Tams1 enzyme-linked immunosorbent assay for diagnosis of Theileria annulata infection in cattle. AB - An enzyme-linked immunosorbent assay (ELISA) was developed based on a recombinant major Theileria annulata merozoite surface antigen, Tams1. Four different recombinant proteins derived from two different Tams1 alleles, both in two different truncated forms, were tested for their performance in the ELISA. Furthermore, antigen concentration, various buffers, washing protocol, and the choice of anti-total-immunoglobulin G (IgG), anti-IgG1, or anti-IgG2 as second antibody were evaluated. The performance of the resulting ELISA was analyzed by measuring the coefficient of variation (CV). A total of 22 sera were analyzed over the measurement range, resulting in a CV of ca. 10%, whereas 30% variation is the maximum acceptable. The cutoff value was determined by the two-graph receiver operating characteristic (TG-ROC), using the indirect fluorescent antibody test (IFAT) as a reference. It was shown that up to 3 months postinfection (p.i.) IFAT is more sensitive and specific, whereas beyond 3 months p.i. ELISA performed as well as IFAT. The cutoff was determined at maximal sensitivity, based on the TG-ROC after 3 months p.i. Nine calves experimentally infected with four different T. annulata stocks remained positive in the ELISA for at least 1 year p.i. Finally, limited cross-reaction was found only with T. parva antisera, but not with any other Theileria or Babesia species. Since the T. parva endemic area hardly overlaps with T. annulata, the Tams1 ELISA has the potential to become a useful tool in the epidemiology of tropical theileriosis. PMID- 10799454 TI - Neutralization profiles of sera from human immunodeficiency virus (HIV)-infected individuals: relationship to HIV viral load and CD4 cell count. AB - The relationship of the neutralizing activity (NA) profile of sera from human immunodeficiency virus (HIV)-infected individuals to the HIV viral load and the absolute CD4 count was examined. The NA of 24 serum samples against autologous isolates (AI) and HIV type 1 strain MN was examined. Three NA patterns were recognized. Nine sera neutralized both AI and MN (+/+), six sera neutralized MN but not AI (-/+), and nine sera failed to neutralize both AI and MN (-/-). The identification of the three neutralization patterns (+/+, -/+, and -/-) indicated that resistance to neutralization was progressive. A reciprocal relationship between the viral burden of the patients and the NA profiles was observed. The nine subjects with a -/- NA profile had a plasma viral load of > or =5 x 10(4) copies/ml and a cellular viral burden of > or =1,122 infectious units per million viable cells, which were significantly different from those of the other groups (P < 0.02). These patterns were independent of the phenotypic characteristics of the virus. Longitudinally, subjects with a -/- profile at baseline gained their HIV-specific NA by 24 weeks of antiretroviral therapy when this was associated with a >/=1-log(10) decline in the plasma HIV viral load. The sera from week 24 from some patients were able to neutralize both the 24-week and the baseline dominant virus isolates. A change in CD4 cell count of 50 or more in either direction predicted a -/- or +/+ profile. The verification of the autologous NA profile might be important in selecting patients who may benefit from immune based therapies involving neutralizing monoclonal antibodies. PMID- 10799455 TI - Treponema pallidum surface immunofluorescence assay for serologic diagnosis of syphilis. AB - A surface immunofluorescence assay (SIFA) using live spirochetes was analyzed and compared with Western blot (WB), fluorescent treponemal antibody absorption (FTA ABS), microhemagglutination (MHA-TP), and Treponema pallidum immobilization (TPI) assays for detecting serum antibodies to T. pallidum in patients with syphilis, in disease controls, and in healthy subjects. SIFA and WB were 99% sensitive (99 of 100 positive specimens) and specific (140 of 140 negative specimens); FTA-ABS showed a sensitivity and a specificity of 90 and 89% (90 of 100 positive and 125 of 140 negative specimens), respectively. MHA-TP showed a sensitivity of 84% (84 of 100 positive specimens) and a specificity of 98.5% (138 of 140 negative specimens). Finally, TPI had a sensitivity of 52% (52 of 100 positive specimens) and a specificity of 100% (140 of 140 negative specimens). The T. pallidum SIFA was therefore highly specific, showing no equivocal reactivities with control sera, and sensitive. The results suggest the possible use of SIFA as a confirmatory test in the serologic diagnosis of syphilis. PMID- 10799456 TI - Comparison of five commercial enzyme-linked immunosorbent assays for detection of antibodies to Bordetella pertussis. AB - Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme linked immunosorbent assays (ELISAs). We compared the performance of five commercially available ELISA kits with the help of 65 serum specimens which were repetitively tested for evaluation of the kits. The specimens contained 20 paired serum samples from patients with clinical pertussis, 15 samples were from children vaccinated with a diphtheria-tetanus-acellular pertussis vaccine, seven specimens were taken from an interlaboratory comparison of ELISAs, and there were three reference preparations from the Food and Drug Administration's (FDA's) Laboratory of Pertussis and from our laboratory. Reference values were obtained from the FDA or from results obtained with an in-house ELISA. Commercial ELISAs were compared with respect to their reproducibility and variability, their ability to detect significant titer rises in paired serum samples, their ability to detect an immune response after vaccination, and the comparability of semiquantitative and quantitative results. Reproducibility was generally good (>89%), intra-assay variation ranged from 2.4 to 28.7%, and indeterminate results were recorded in up to 18.5% of all specimens. Most kits correctly identified the antibody response to an acellular pertussis vaccine. None of the commercial kits identified all cases of pertussis correctly, and the sensitivity ranged between 60 and 95%. All five commercial ELISAs showed great discrepancies when comparing semiquantitative results and contained obviously different antigen preparations. Our data suggest that the five commercial ELISAs tested here need further improvement and standardization. PMID- 10799457 TI - New immunofluorescence assays for detection of Human herpesvirus 8-specific antibodies. AB - Several assays have been developed for detection of immunoglobulin G antibodies to Human herpesvirus 8 (HHV-8), including immunofluorescence assays (IFAs) and enzyme-linked immunosorbent assays (ELISAs). However, the specificity and sensitivity of these assays are not completely defined due to the lack of a "gold standard." Although IFAs based on primary effusion lymphoma (PEL) cell lines are used widely, the assays can be confounded by nonspecific reactions against cellular components and potential cross-reaction with antibodies against other herpesviruses. To provide more reliable IFAs, we established recombinant Semliki Forest viruses (rSFVs) expressing the HHV-8-specific proteins ORF73 and K8.1 and used BHK-21 cells infected with these rSFVs for IFA (ORF73-IFA and K8.1-IFA). Expression of the HHV-8-specific proteins at very high levels by the rSFV system allowed easy scoring for IFA and thereby increased specificity. The rSFV system also allowed detection of antibodies against glycosylation-dependent epitopes of K8.1. Titers measured by rSFV-based IFAs and PEL-based IFAs correlated well (correlation coefficients of >0.9), and concordances of seroreactivities between rSFV-based and PEL-based IFAs were >97% (kappa > 0.93). K8.1-IFA was more sensitive than either ORF73-IFA or peptide ELISAs. Using PEL-based lytic IFA as a reference assay, the sensitivity and specificity of K8.1-IFA were estimated to be 94 and 100%, respectively. HHV-8 prevalences determined by K8.1-IFA among the human immunodeficiency virus (HIV)-positive (HIV(+)) Kaposi's sarcoma (KS) patients, HIV(+) KS(-) patients, and healthy controls were 100, 65, and 6.7%, respectively, which were consistent with prior reports. Therefore, our rSFV-based IFAs may provide a specific and sensitive method for use in epidemiology studies. In addition, they will provide a basis for further development of diagnostic tests for HHV-8 infection. PMID- 10799458 TI - Development of the antinuclear and anticytoplasmic antibody consensus panel by the Association of Medical Laboratory Immunologists. AB - The Association of Medical Laboratory Immunologists (AMLI) have developed a panel of antinuclear and anticytoplasmic antibody consensus sera that can be useful for enzyme immunoassay (EIA), Ouchterlony, and immunofluorescence assay methods. It was developed to assist in the evaluation of newly available EIA methods for the detection of autoantibodies. The panel of sera was evaluated in several clinical laboratories and a large number of laboratories owned by manufacturers of clinical autoantibody testing kits. The majority of sera performed well for the EIAs in both the clinical laboratories and the manufacturers' laboratories, but some samples had discrepant results. A major source of discrepancy is the current inability of the EIA results to be directly compared in a quantitative way as no standardization exists. The evaluation demonstrated lower sensitivity of detection by the Ouchterlony method. The limited evaluation of the sera with immunoblotting and Western blotting did not show good agreement with other methods. Further work must be done to standardize blotting methods prior to their use in routine clinical testing. The sera are now available to vendors and clinical laboratories for use in the detection of SS-A, SS-B, Sm, U1-RNP, Scl-70, Jo-1, double-stranded DNA, and centromere antibodies. The availability of the consensus sera will help evaluate and improve the EIA methods currently being used. PMID- 10799459 TI - Susceptibility of Vibrio cholerae O139 to antibody-dependent, complement-mediated bacteriolysis. AB - Volunteer studies with Vibrio cholerae O1 have shown that the best correlate of a vaccine's protective efficacy is its propensity to elicit serum bactericidal responses in its recipients. Attempts to detect such responses following infection with V. cholerae O139, however, have met with varying success. Using a tube-based assay which involves viable counting, we now report that strains of serogroup O139 can appear to be sensitive or resistant to a fixed concentration of complement in the presence of antibody, depending on assay conditions. Susceptibility to lysis is critically dependent on the availability of complement, but with O139 indicator strains this is not simply determined by the concentration of serum added to the reaction mix. The nature of the assay diluent and the concentration of indicator bacteria can also dramatically affect bactericidal end points, whereas such variables have minimal significance with O1 indicator bacteria. Although some laboratories use unencapsulated mutant strains to seek evidence of seroconversion following exposure to V. cholerae O139, this is not necessary, and our findings question the significance of capsule expression as a determinant of complement sensitivity when antibody is present. The medium used for growth of the indicator strain and the particular strain used appeared to be unimportant. Each of seven O139 isolates tested was found to be lysed by antibody and complement in our standard assay system, which allowed the detection of significant serum bactericidal responses in 9 of 11 cases of O139 disease. PMID- 10799460 TI - Evaluation of 12 commercial tests for detection of Epstein-Barr virus-specific and heterophile antibodies. AB - Ten microbiological departments in Norway have participated in a multicenter evaluation of the following commercial tests for detection of Epstein-Barr virus (EBV)-specific and heterophile antibodies: CAPTIA Select viral capsid antigen (VCA)-M/G/EBNA (Centocor Inc.), Enzygnost anti-EBV/immunoglobulin M (IgM) and IgG (Dade Behring), Vironostika EBV VCA IgM/IgG/EBNA enzyme-linked immunosorbent assay (ELISA) (Organon Teknika), SEROFLUOR immunofluorescence assay and EBV Combi Test (Institute Virion Ltd.), anti-EBV recombinant IgM- and IgG-early antigen/EBNA IgG ELISA (Biotest Diagnostics), EBV IgM/IgG/EBNA ELISA (Gull Laboratories), Paul-Bunnell-Davidsohn test (Sanofi Diagnostics Pasteur), Monosticon Dri-Dot (Organon Teknika), Avitex-IM (Omega Diagnostics Ltd.), Alexon Serascan infectious mononucleosis test (Alexon Biomedical Inc. ), Clearview IM (Unipath Ltd.), and Cards+/-OS Mono (Pacific Biotech, Inc.). The test panel included sera from patients with primary EBV infection, immunocompromised patients with recent cytomegalovirus infection, healthy persons (blood donors), and EBV-seronegative persons. Among the tests for EBV-specific antibodies the sensitivity was good, with only small differences between the different assays. However, there was a greater variation in specificity, which varied between 100% (Enzygnost) and 86% (Biotest). Tests for detection of heterophile antibodies based on purified or selected antigen (Avitex, Alexon, Clearview IM, and Cards+/ OS Mono) were more sensitive than the Paul-Bunnell-Davidsohn and Monosticon tests. PMID- 10799461 TI - Conjugation of hydroxyethyl starch to desferrioxamine (DFO) modulates the dual role of DFO in Yersinia enterocolitica infection. AB - The iron chelator desferrioxamine (DFO) B is widely used in the therapy of patients with iron overload. As a side effect, DFO may favor the occurrence of fulminant Yersinia infections. Previous work from our laboratory showed that this might be due to a dual role of DFO: growth promotion of the pathogen and immunosuppression of the host. In this study, we sought to determine whether conjugation of DFO to hydroxyethyl starch (HES-DFO) may prevent exacerbation of Yersinia infection in mice. We found HES-DFO to promote neither growth of Yersinia enterocolitica nor mitogen-induced T-cell proliferation and gamma interferon production by T cells in vitro. Nevertheless, in vivo HES-DFO promoted growth of Y. enterocolitica possibly due to cleavage of HES and release of DFO. The pretreatment of mice with DFO resulted in death of all mice 2 to 5 days after application of a normally sublethal inoculum of Y. enterocolitica, while none of the mice pretreated with HES-DFO died within the first 7 days postinfection. However, some of the HES-DFO-treated mice died 8 to 14 days postinfection. Thus, due to the delayed in vivo effect HES-DFO failed to trigger Yersinia-induced septic shock, which accounts for early mortality in DFO-associated septicemia. Moreover, our data suggest that DFO needs to be taken up by host cells in order to exert its immunosuppressive action. These results strongly suggest that HES DFO might be a favorable drug with fewer side effects than DFO in terms of DFO promoted fulminant infections. PMID- 10799462 TI - Antibody response of patients with Helicobacter pylori-related gastric adenocarcinoma: significance of anti-cagA antibodies. AB - The aim of this study was to search for a specific antibody pattern in sera from patients suffering from Helicobacter pylori-related gastric adenocarcinoma (GAC). The serological response of 22 patients suffering from GAC, 31 patients with gastroduodenal ulcer, and 39 asymptomatic subjects was analyzed using immunoblotting performed with three H. pylori strains: strain ATCC 43579; strain B110, isolated from a patient with ulcers; and strain B225, isolated from a patient with GAC. In addition, the latex agglutination test Pyloriset Dry was used to analyze ambiguous sera. H. pylori seropositivity was 75% in the GAC group, 61.3% in the ulcer group, and 56.4% in the asymptomatic group. Anti-CagA antibodies were found more often in the GAC group (48.8%) and in the ulcer group (47.3%) than in the asymptomatic group (21.2%). These percentages depended on the strain used as an antigen: in the GAC group, the anti-CagA frequencies were 93.3, 40, and 13.3% with strains B225, B110, and ATCC 43579, respectively. Thus the presence of anti-CagA antibodies was increased in patients suffering from H. pylori-related GAC, in particular when the CagA antigen was from a GAC strain. These data suggest the existence of a CagA protein specifically expressed by H. pylori strains isolated from GAC patients. PMID- 10799463 TI - Are the enzyme immunoassays for antibodies to pneumococcal capsular polysaccharides serotype specific? AB - The specificity of antibody binding to pneumococcal capsular polysaccharides (Pnc PSs) measured by enzyme immunoassay (EIA) was studied by inhibition of antibody binding by homologous and heterologous PSs. We found extensive cross-reactivity of antibody binding to type 6B, 19F, and 23F PSs but not to type 14 PS, even after treatment with cell wall PS (CPS). The cross-reactive antibody was highly prevalent in sera of infants and adults with naturally acquired antibody, but not in sera of infants and adults immunized with pneumococcal vaccines. However, a type 11A antibody response was seen after vaccination with heterologous PSs. Monoclonal antibodies prepared against a type 6B PS-tetanus toxoid conjugate recognized also other than the specific type of PS in the EIA, implying the possible existence of a cross-reactive epitope. Remarkable differences in specificity among type 6B PS preparations from different manufacturers were found. Moreover, different lots of type 11A PS from the same manufacturer showed differences in specificity. The results suggest that some Pnc PS preparations may contain cross-reactive epitopes or impurities, other than CPS, that are common to many types of Pnc PS. The specificity of antibodies, especially in sera from nonimmunized subjects, measured by EIA can be questioned. PMID- 10799464 TI - Comparison of commercial latex agglutination and sandwich enzyme immunoassays with a competitive binding inhibition enzyme immunoassay for detection of antigenemia and antigenuria in a rabbit model of invasive aspergillosis. AB - A commercial latex agglutination assay (LA) and a sandwich enzyme immunoassay (SEIA) (Sanofi Diagnostics Pasteur, Marnes-la-Coquette, France) were compared with a competitive binding inhibition assay (enzyme immunoassay [EIA]) to determine the potential uses and limitations of these antigen detection tests for the sensitive, specific, and rapid diagnosis of invasive aspergillosis (IA). Toward this end, well-characterized serum and urine specimens were obtained by using a rabbit model of IA. Serially collected serum or urine specimens were obtained daily from control rabbits or from rabbits immunosuppressed and infected systemically with Aspergillus fumigatus. By 4 days after infection, EIA, LA, and SEIA detected antigen in the sera of 93, 93, and 100% of A. fumigatus-infected rabbits, respectively, whereas antigen was detected in the urine of 93, 100, and 100% of the rabbits, respectively. False-positive results for non-A. fumigatus infected rabbits for EIA, LA, and SEIA were as follows: for serum, 14, 11, and 23%, respectively; for urine, 14, 84, and 90%, respectively. Therefore, although the sensitivities of all three tests were similar, the specificity was generally greater for EIA than for LA or SEIA. Infection was also detected earlier by EIA, by which the serum of 53% of A. fumigatus-infected rabbits was positive as early as 1 day after infection, whereas the serum of only 27% of the rabbits tested by LA was positive. Although the serum of 92% of A. fumigatus-infected rabbits was positive by SEIA as early as 1 day after infection, the serum of a high percentage (50%) was false positive before infection. The urine of 21% of A. fumigatus-infected rabbits was positive by EIA as early as 1 day after infection, and the urine of none of the rabbits was false positive before infection. When EIA results for urine specimens were combined with those for serum, sensitivity was improved (i.e., 67% of rabbits were positive by 1 day after infection and only one rabbit gave a false-positive result). A total of 93% of A. fumigatus infected rabbits were positive for antigen in urine as early as 1 day after infection and the urine of 100% of the rabbits was positive by SEIA. However, before infection, 79% of A. fumigatus-infected rabbits were false positive for antigen in urine by LA and 90% were false positive for antigen in urine by SEIA. These data indicate that the EIA has the potential to be used to diagnose IA with both serum and urine specimens and to detect a greater number of infections earlier with greater specificity than the specificities achieved with the commercial tests. PMID- 10799465 TI - A latex bead-based flow cytometric immunoassay capable of simultaneous typing of multiple pneumococcal serotypes (Multibead assay). AB - A simple and rapid method of simultaneously determining 15 Streptococcus pneumoniae serotypes was developed. Fifteen latex beads of different sizes and different red fluorescence levels were coated with 1 of 15 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9N, 9V, 14, 18C, 19A, 19F, 22F, and 23F) of pneumococcal capsular polysaccharide (PS). The bead mixture was incubated with individual pneumococcal lysate, a pool of rabbit antisera capable of binding the 15 serotypes, and fluorescein (green fluorescence)-conjugated anti-rabbit antibody. Bead size, red fluorescence, and green fluorescence were measured in a single flow cytometer run. The green fluorescence of the beads was inhibited only when there was a serotypic match between PS on the bead and PS in the pneumococcal lysate. This method distinguished cross-reactive serotypes and correctly identified the serotypes in 100% of 86 pneumococcal isolates tested. PMID- 10799466 TI - Specificity and prevalence of natural bovine anti-alpha galactosyl (Galalpha1 6Glc or Galalpha1-6Gal) antibodies. AB - Immunity against the carbohydrate components of microorganisms mediated by antibodies is an important part of host defenses. Humans and closely related primates, but not other mammals, possess natural anti-Galalpha1-3Gal antibodies which also, although less avidly, react with melibiose (Galalpha1-6Glc). Using an enzyme-linked immunosorbent assay (ELISA) with melibiose-bovine serum albumin as an antigen, we analyzed bovine anti-alpha galactosyl antibodies with respect to specificity and distribution in individual animals. Inhibition assays showed that melibiose was the strongest inhibitor, followed equally by stachyose (Galalpha1 6Galalpha1-6Glcbeta1-2Fru) and raffinose (Galalpha1-6Glcbeta1-2Fru) and then by Galbeta1-6Gal, Gal, and Galalpha1-2Gal. Others, including Galalpha1-3Gal and Galalpha1-4Gal, only exhibited minor inhibition. Thus, these bovine anti-alpha galactosyl antibodies appeared to preferentially react with Galalpha1-6Glc or Galalpha1-6Gal. The distinction of this specificity from that (Galalpha1-3Gal) of human antibodies was further demonstrated by the poor reaction of bovine serum to the Galalpha1-3Gal antigen in comparison to human serum. All 27 healthy bovine serum samples of the three age groups (newborn, calf, and adult) tested contained such antibodies with titers increasing with age. The antibodies purified by affinity chromatography using a melibiose-agarose column were mainly of the immunoglobulin G (IgG) isotype with a concentration of >23 microg/ml in most samples. IgG1 was found to be the primary antimelibiose IgG isotype in all age groups by isotype-specific ELISA, but a significant increase in IgG2, an isotype more related to innate immunity, was observed in calves and adults, compared to newborns. The purified antibodies reacted with the type II bovine strain of Streptococcus agalactiae, a common pathogen of bovine mastitis. Thus, these anti Galalpha1-6Glc or Galalpha1-6Gal antibodies in cattle might be involved in defense against microbes bearing this or the related epitopes. PMID- 10799467 TI - Humoral immune response in chromoblastomycosis during and after therapy. AB - A longitudinal study was carried out in Madagascar, the most important focus of chromoblastomycosis (P. Esterre, A. Andriantsimahavandy, E. Ramarcel, and J. L. Pecarrere, Am. J. Trop. Med. Hyg. 55:45-47, 1996), to investigate natural immunity to this disease. Sequential blood samples were obtained before, during, and at the end of a successful therapeutic trial with terbinafine, a new antifungal drug. Using enzyme-linked immunosorbent assay and immunoblot methods, detailed analyses of antibody concentration and antigen mapping were conducted for 136 serum samples and tentatively correlated to epidemiological and pathobiological data. Two different cytoplasmic antigens, corresponding to the two fungal species involved (Fonsecaea pedrosoi and Cladophialophora carrionii), were used to analyze the distribution of different classes of immunoglobulins. This was done with respect to the origin of the isolates, clinical and pathobiological. Although strong individual variations were noticed, some major antigens (one of 18.5 kDa specific for F. pedrosoi and two of 23.5 and 33 kDa, respectively, specific for C. carrionii) corresponded to high antibody prevalence and concentration. As some antigenic components were also detected by immunoglobulin M (IgM) and IgA antibodies, the role that these specific antibodies could play in the immune response is discussed. PMID- 10799468 TI - Intestinal secretory immunoglobulin A response to enteroaggregative Escherichia coli in travelers with diarrhea. AB - We examined stool samples from travelers for secretory immunoglobulin A (sIgA) to enteroaggregative Escherichia coli (EAEC) during episodes of acute diarrhea. Ten paired samples from 10 patients with diarrhea caused by EAEC were examined for the presence of specific sIgA by dot blot and Western blot immunoassays. Five samples were positive by dot blotting, and two samples were positive by Western blotting. PMID- 10799469 TI - Diagnosis of Enterocytozoon bieneusi by PCR in stool samples eluted from filter paper disks. AB - We report a PCR-based assay for the detection of Enterocytozoon bieneusi. We extracted DNA from feces which had been applied to filter paper disks and evaluated four preserving solutions. Infected specimens were identified by electrophoresis of amplicons from concentrated formalin-fixed samples and unconcentrated fresh feces. Our findings demonstrate that this methodology is effective for sample collection, mailing, and diagnosis of this pathogen. PMID- 10799470 TI - Clonal analysis of the human B-cell repertoire using a heteromobility assay. AB - A heteromobility duplex tracking assay was developed to analyze B-cell clonality. The assay was based on the genetic variability of B-cell immunoglobulin (Ig) sequences. Binding of amplified (Ig) sequences to a single-stranded radiolabeled Ig DNA probe resulted in the formation of heteroduplexes. The mobilities of these heteroduplexes helped to distinguish clonal B cells. PMID- 10799471 TI - Apoptosis of primary-culture rat microglial cells induced by pathogenic Acanthamoeba spp. AB - To determine whether trophozoites and lysates of pathogenic Acanthamoeba spp. induce apoptosis in primary-culture microglial cells, transmission electron microscopic (TEM) examinations, assessment of DNA fragmentation by agarose gel electrophoresis, and the TdT-mediated dUTP nick-end labeling assay were performed. When a trophozoite of pathogenic Acanthamoeba culbertsoni came in contact with a microglial cell, the digipodium was observed by TEM. Nuclear chromatin condensation was observed in 10% of microglial cells, while it was not revealed when they were cocultured with weakly pathogenic Acanthamoeba royreba trophozoites. DNA fragmentation in microglial cells cocultured with the A. culbertsoni lysate was detected by electrophoresis, showing DNA ladder formation, whereas it was hardly observed in microglial cells cocultured with A. royreba. DNA fragmentation of microglial cells was also confirmed by flow cytometry analysis. The fluorescence of TdT-stained apoptotic bodies became intensely visible with microglial cells cocultured with the A. culbertsoni lysate. In contrast, with microglial cells cocultured with the A. royreba lysate, only a background level of fluorescence of TdT-stained apoptotic bodies was detected. These results suggest that some rat microglial cells cocultured with pathogenic A. culbertsoni undergo cytopathic changes which show the characteristics of the apoptotic process, such as nuclear condensation and DNA fragmentation. PMID- 10799472 TI - Secretory leukocyte protease inhibitor inhibits infection of monocytes and lymphocytes with human immunodeficiency virus type 1 but does not interfere with transcytosis of cell-associated virus across tight epithelial barriers. AB - In the present study, we demonstrate that recombinant human secretory leukocyte protease inhibitor (rhSLPI) inhibits infection of lymphocyte- and monocyte derived tumor cell lines and peripheral blood lymphocytes with laboratory-adapted isolates and with the primary isolate, NDK, of free human immunodeficiency virus type 1 (HIV-1). In contrast, rhSLPI did not exhibit inhibitory activity toward transcytosis of cell-associated HIV-1 through a tight monolayer of endometrial epithelial cells. These observations indicate that the inhibitory effect of SLPI is restricted to free HIV-1 in corporal fluids. PMID- 10799474 TI - Defining the carbohydrate specificities of aplysia gonad lectin exhibiting a peculiar D-galacturonic acid affinity. AB - Aplysia gonad lectin (AGL), which has been shown to stimulate mitogenesis in human peripheral lymphocytes, to suppress tumor cells, and to induce neurite outgrowth and improve cell viability in cultured Aplysia neurons, exhibits a peculiar galacturonic acid/galactose specificity. The carbohydrate binding site of this lectin was characterized by enzyme-linked lectino-sorbent assay and by inhibition of AGL-glycan interactions. Examination of the lectin binding with 34 glycans revealed that it reacted strongly with the following glycoforms: most human blood group precursor (equivalent) glycoproteins (gps), two Galalpha1- >4Gal-containing gps, and two d-galacturonic acid (GalUA)-containing polysaccharides (pectins from apple and citrus fruits), but poorly with most human blood group A and H active and sialylated gps. Among the GalUA and mammalian saccharides tested for inhibition of AGL-glycan binding, GalUA mono- to trisaccharides were the most potent ones. They were 8.5 x 10(4) times more active than Gal and about 1.5 x 10(3) more active than the human blood group P(k) active disaccharide (E, Galalpha1-->4Gal). This disaccharide was 6, 28, and 120 times more efficient than Galbeta1-->3GlcNAc(I), Galbeta1-->3GalNAc(T), and Galbeta1--> 4GlcNAc (II), respectively, and 35 and 80 times more active than melibiose (Galalpha1-->6Glc) and human blood group B active disaccharide (Galalpha1- >3Gal), respectively, showing that the decreasing order of the lectin affinity toward alpha-anomers of Gal is alpha1-->4 > alpha1-->6 > alpha1-->3. From the data provided, the carbohydrate specificity of AGL can be defined as GalUAalpha1- >4 trisaccharides to mono GalUA > branched or cluster forms of E, I, and II monomeric E, I, and II, whereas GalNAc is inactive. PMID- 10799473 TI - Monoclonal antibody differentiation of Mycoplasma mycoides subsp. mycoides small colony strains causing contagious bovine pleuropneumonia from less important large-colony strains. AB - Monoclonal antibody (MAb) PK-2 inhibited the in vitro growth of nine Mycoplasma mycoides subsp. mycoides small-colony strains. In contrast to the results with polyclonal antisera, growth inhibition by MAb PK-2 was specific for M. mycoides subsp. mycoides small-colony strains and constituted a reliable means of distinguishing them from other mycoplasmas. PMID- 10799475 TI - Protein traffic from the secretory pathway to the endosomal system in pancreatic beta-cells. AB - Constitutive-like secretion involves vesicular trafficking corresponding kinetically and biochemically with a post-trans-Golgi network (TGN) origin. In pancreatic beta-cells, the budding of AP-1/clathrin-coated vesicles, a portion of which is derived from immature secretory granules, has been hypothesized to initiate constitutive-like trafficking. However, approximately 30 min after release of a 20 degrees C intracellular transport block in pancreatic beta-cells (to synchronize protein egress from the TGN), addition of brefeldin A (BFA) (which inhibits AP-1 recruitment) was reported not to block subsequent constitutive-like secretion. To further explore post-TGN trafficking in pancreatic beta-cell lines, we have followed the fate of pulse-labeled procathepsin B (ProB, a lysosomal proenyzme) after postpulse wortmannin treatment or the BFA treatment described above. We find that continuous wortmannin treatment allows ProB to reach immature secretory granules but inhibits its egress from maturing granules. Remarkably, BFA treatment causes augmented unstimulated secretion of newly synthesized ProB that is not paralleled by insulin. This effect requires a delay of 25-35 min after release from the 20 degrees C block. Further, when ProB delivery to endosomes is inhibited, its BFA augmented secretion is eliminated. We hypothesize that the constitutive-like pathway involves an endosomal intermediate. PMID- 10799476 TI - Two glyceraldehyde-3-phosphate dehydrogenases with opposite physiological roles in a nonphotosynthetic bacterium. AB - Bacillus subtilis possesses two similar putative phosphorylating glyceraldehyde-3 phosphate dehydrogenase (GAPDH) encoding genes, gap (renamed gapA) and gapB. A gapA mutant was unable to grow on glycolytic carbon sources, although it developed as well as the wild-type strain on gluconeogenic carbon sources. A gapB mutant showed the opposite phenotype. Purified GapB showed a 50-fold higher GAPDHase activity with NADP(+) than with NAD(+), with K(m) values of 0.86 and 5.7 mm, respectively. lacZ reporter gene fusions revealed that the gapB gene is transcribed during gluconeogenesis and repressed during glycolysis. Conversely, gapA transcription is 5-fold higher under glycolytic conditions than during gluconeogenesis. GAPDH activity assays in crude extracts of wild-type and mutant strains confirmed this differential expression pattern at the enzymatic level. Genetic analyses demonstrated that gapA transcription is repressed by the yvbQ (renamed cggR) gene product and indirectly stimulated by CcpA. Thus, the same enzymatic step is catalyzed in B. subtilis by two enzymes specialized, through the regulation of their synthesis and their enzymatic characteristics, either in catabolism (GapA) or in anabolism (GapB). Such a dual enzymatic system for this step of the central carbon metabolism is described for the first time in a nonphotosynthetic eubacterium, but genomic analyses suggest that it could be a widespread feature. PMID- 10799477 TI - Bicarbonate enhances the hydroxylation, nitration, and peroxidation reactions catalyzed by copper, zinc superoxide dismutase. Intermediacy of carbonate anion radical. AB - The effect of bicarbonate anion (HCO(3)(-)) on the peroxidase activity of copper, zinc superoxide dismutase (SOD1) was investigated using three structurally different probes: 5, 5'-dimethyl-1-pyrroline N-oxide (DMPO), tyrosine, and 2, 2' azino-bis-[3-ethylbenzothiazoline]-6-sulfonic acid (ABTS). Results indicate that HCO(3)(-) enhanced SOD/H(2)O(2)-dependent (i) hydroxylation of DMPO to DMPO-OH as measured by electron spin resonance, (ii) oxidation and nitration of tyrosine to dityrosine, nitrotyrosine, and nitrodityrosine as measured by high pressure liquid chromatography, and (iii) oxidation of ABTS to the ABTS cation radical as measured by UV-visible spectroscopy. Using oxygen-17-labeled water, it was determined that the oxygen atom present in the DMPO-OH adduct originated from H(2)O and not from H(2)O(2). This result proves that neither free hydroxyl radical nor enzyme-bound hydroxyl radical was involved in the hydroxylation of DMPO. We postulate that HCO(3)(-) enhances SOD1 peroxidase activity via formation of a putative carbonate radical anion. This new and different perspective on HCO(3)(-)-mediated oxidative reactions of SOD1 may help us understand the free radical mechanism of SOD1 and related mutants linked to amyotrophic lateral sclerosis. PMID- 10799478 TI - Membrane type 4 matrix metalloproteinase (MMP17) has tumor necrosis factor-alpha convertase activity but does not activate pro-MMP2. AB - Membrane type 4 matrix metalloproteinase (MT4-MMP) shows the least sequence homology to the other MT-MMPs, suggesting a distinct function for this protein. We have isolated a complete cDNA corresponding to the mouse homologue which includes the signal peptide and a complete pro-domain, features that were lacking from the human form originally isolated. Mouse MT4-MMP (mMT4-MMP) expressed in COS-7 cells is located at the cell surface but does not show ability to activate pro-MMP2. The pro-catalytic domain was expressed in Escherichia coli as insoluble inclusions and active enzyme recovered after refolding. Activity of the isolated catalytic domain against synthetic peptides commonly used for MMP enzyme assays could be inhibited by TIMP1, -2, and -3. The recombinant mMT4-MMP catalytic domain was also unable to activate pro-MMP2 and was very poor at hydrolyzing components of the extracellular matrix with the exception of fibrinogen and fibrin. mMT4-MMP was able to hydrolyze efficiently a peptide consisting of the pro-tumor necrosis factor alpha (TNFalpha) cleavage site, a glutathione S transferase-pro-TNFalpha fusion protein, and was found to shed pro-TNFalpha when co-transfected in COS-7 cells. MT4-MMP was detected by Western blot in monocyte/macrophage cell lines which in combination with its fibrinolytic and TNFalpha-converting activity suggests a role in inflammation. PMID- 10799479 TI - Polyglutamylation of folate coenzymes is necessary for methionine biosynthesis and maintenance of intact mitochondrial genome in Saccharomyces cerevisiae. AB - One-carbon metabolism is essential to provide activated one-carbon units in the biosynthesis of methionine, purines, and thymidylate. The major forms of folates in vivo are polyglutamylated derivatives. In organisms that synthesize folate coenzymes de novo, the addition of the glutamyl side chains is achieved by the action of two enzymes, dihydrofolate synthetase and folylpolyglutamate synthetase. We report here the characterization and molecular analysis of the two glutamate-adding enzymes of Saccharomyces cerevisiae. We show that dihydrofolate synthetase catalyzing the binding of the first glutamyl side chain to dihydropteroate yielding dihydrofolate is encoded by the YMR113w gene that we propose to rename FOL3. Mutant cells bearing a fol3 mutation require folinic acid for growth and have no dihydrofolate synthetase activity. We show also that folylpolyglutamate synthetase, which catalyzes the extension of the glutamate chains of the folate coenzymes, is encoded by the MET7 gene. Folylpolyglutamate synthetase activity is required for methionine synthesis and for maintenance of mitochondrial DNA. We have tested whether two folylpolyglutamate synthetases could be encoded by the MET7 gene, by the use of alternative initiation codons. Our results show that the loss of mitochondrial functions in met7 mutant cells is not because of the absence of a mitochondrial folylpolyglutamate synthetase. PMID- 10799480 TI - Mitochondrial aconitase is a source of hydroxyl radical. An electron spin resonance investigation. AB - Mitochondrial aconitase (m-aconitase) contains a [4Fe-4S](2+) cluster in its active site that catalyzes the stereospecific dehydration-rehydration of citrate to isocitrate in the Krebs cycle. It has been proposed that the [4Fe-4S](2+) aconitase is oxidized by superoxide, generating the inactive [3Fe-4S](1+) aconitase. In this reaction, the likely products are iron(II) and hydrogen peroxide. Consequently, the inactivation of m-aconitase by superoxide may increase the formation of hydroxyl radical ((*)OH) through the Fenton reaction in mitochondria. In this work, evidence for the generation of (*)OH from the reaction of m-aconitase with superoxide is provided using ESR spin trapping experiments with 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide and alpha phenyl-N-tert-butylnitrone. Formation of free ( small middle dot)OH was verified with the (*)OH scavenger Me(2)SO, which forms methyl radical upon reacting with (*)OH. The addition of Me(2)SO to incubation mixtures containing m-aconitase and xanthine/xanthine oxidase yielded methyl radical, which was detected by ESR spin trapping. Methyl radical formation was further confirmed using [(13)C]Me(2)SO. Parallel low temperature ESR experiments demonstrated that the generation of the [3Fe-4S](1+) cluster increased with increasing additions of superoxide to m aconitase. This reaction was reversible, as >90% of the initial aconitase activity was recovered upon treatment with glutathione and iron(II). This mechanism presents a scenario in which (*)OH may be continuously generated in the mitochondria. PMID- 10799481 TI - Cu2+ and Zn2+ inhibit nitric-oxide synthase through an interaction with the reductase domain. AB - Cu(2+) and Zn(2+) inhibit all of the NADPH-dependent reactions catalyzed by neuronal nitric-oxide synthase (nNOS) including ferricytochrome c reduction, NADPH oxidation, and citrulline formation. Cu(2+) and Zn(2+) also inhibit ferricytochrome c reduction by the independent reductase domain. Zn(2+) affects all activities of the full-length nNOS and the reductase domain to the same extent (estimated IC(50) values from 9 to 31 microm), suggesting Zn(2+) occupation of a single site in the reductase domain. Citrulline formation and NADPH oxidation by the full-length nNOS and ferricytochrome c reduction by the reductase domain are affected similarly by Cu(2+), with estimated IC(50) values ranging from 6 to 33 microm. However, Cu(2+) inhibits ferricytochrome c reduction by the full-length nNOS 2 orders of magnitude more potently, with an estimated IC(50) value of 0.12 microm. These data suggest the possibility that Cu(2+) may interact with nNOS at two sites, one composed exclusively of the reductase domain (which is perhaps also involved in Zn(2+)-mediated inhibition), and another that includes components of both domains. Occupation of the second (higher affinity) site could then promote the selective inhibition of ferricytochrome c reduction in full-length nNOS. Neither the inhibition by Cu(2+) nor that by Zn(2+) is dependent on calmodulin. PMID- 10799483 TI - Analysis by high density cDNA arrays of altered gene expression in human intestinal epithelial cells in response to infection with the invasive enteric bacteria Salmonella. AB - Many clinically important enteric pathogens initiate disease by invading and passing through the intestinal epithelium, a process accompanied by increased epithelial expression of proinflammatory cytokines. To further define the role intestinal epithelial cells play in initiating and modulating the host response to infection with invasive bacteria, hybrid selection on high density cDNA arrays was used to characterize the mRNA expression profile of approximately 4,300 genes in human intestinal epithelial cells after infection with the prototypic invasive bacteria, Salmonella. Selected findings were further evaluated by reverse transcription-polymerase chain reaction, Northern blot analysis, and protein assays. Epithelial infection with Salmonella significantly up-regulated mRNA expression of a relatively small fraction of all genes tested. Of these, several cytokines (granulocyte colony-stimulating factor, inhibin A, Epstein-Barr virus induced gene 3, interleukin-8, macrophage inflammatory protein-2alpha), kinases (TKT, Eck, HEK), transcription factors (interferon regulatory factor-1), and HLA class I were the most prominent. Furthermore, the transcription factor NF-kappaB is shown to be important for inducible mRNA expression for a broad group of genes tested. These findings expand the repertoire of known epithelial cell responses to infection with an invasive enteric pathogen. The results also show that evaluation of mRNA expression profiles by cDNA array analysis is a powerful approach to characterizing and understanding host-pathogen interactions. PMID- 10799482 TI - Alzheimer's beta-amyloid, human islet amylin, and prion protein fragment evoke intracellular free calcium elevations by a common mechanism in a hypothalamic GnRH neuronal cell line. AB - A growing number of reports suggest that elevated levels of extracellular Alzheimer's beta-amyloid protein alter the homeostasis of free [Ca(2+)](i) in different cell types of the mammalian brain. In line with these results, we have previously shown that AbetaP[1-40] forms cation-selective channels (Ca(2+) included) across artificial planar bilayers formed from acidic phospholipids and across excised membrane patches from immortalized hypothalamic GnRH neurons (GT1 7 cells), suggesting that the nonregulated Ca(2+)-influx through these spontaneously formed "amyloid channels" may provide a mechanism to explain its toxicity (1). We have now found and report here that the application of AbetaP[1 40] to GT1-7 neurons consistently elevates [Ca(2+)](i) levels. We also found that human islet amylin and the prion protein fragment (PrP106-126), peptides that acquire beta-pleated sheet conformation in water solutions and have been reported to form ion channels across planar bilayer membranes, also increase cytosolic free calcium in GT1-7 neurons. Searching for protective agents, we found that soluble cholesterol, known to decrease the fluidity of the cell membrane, inhibits AbetaP[1-40]-evoked [Ca(2+)](i) rise. These results suggest that unregulated Ca(2+) entry across amyloid channels may be a common mechanism causing cell death, not only in diseases of the third age, including Alzheimer's disease and type 2 diabetes mellitus, but also in prion-induced diseases. PMID- 10799484 TI - cDNA cloning of phosphoethanolamine N-methyltransferase from spinach by complementation in Schizosaccharomyces pombe and characterization of the recombinant enzyme. AB - The N-methylation of phosphoethanolamine is the committing step in choline biogenesis in plants and is catalyzed by S-adenosyl-L methionine:phosphoethanolamine N-methyltransferase (PEAMT, EC ). A spinach PEAMT cDNA was isolated by functional complementation of a Schizosaccharomyces pombe cho2(-) mutant and was shown to encode a protein with PEAMT activity and without ethanolamine- or phosphatidylethanolamine N-methyltransferase activity. The PEAMT cDNA specifies a 494-residue polypeptide comprising two similar, tandem methyltransferase domains, implying that PEAMT arose by gene duplication and fusion. Data base searches suggested that PEAMTs with the same tandem structure are widespread among flowering plants. Size exclusion chromatography of the recombinant enzyme indicates that it exists as a monomer. PEAMT catalyzes not only the first N-methylation of phosphoethanolamine but also the two subsequent N methylations, yielding phosphocholine. Monomethyl- and dimethylphosphoethanolamine are detected as reaction intermediates. A truncated PEAMT lacking the C-terminal methyltransferase domain catalyzes only the first methylation. Phosphocholine inhibits both the wild type and the truncated enzyme, although the latter is less sensitive. Salinization of spinach plants increases PEAMT mRNA abundance and enzyme activity in leaves by about 10-fold, consistent with the high demand in stressed plants for choline to support glycine betaine synthesis. PMID- 10799485 TI - A new SUMO-1-specific protease, SUSP1, that is highly expressed in reproductive organs. AB - A full-length cDNA encoding a SUMO-1-specific protease, named SUSP1, was identified and cloned for the first time from the human brain. Nucleotide sequence analysis of the cDNA containing an open reading frame of 3336 base pairs revealed that the protease consists of 1112 amino acids with a calculated molecular mass of 126,116 Da. Like yeast Ulp1, SUSP1 is a cysteine protease containing the well conserved His/Asp/Cys catalytic triad. SUSP1 expressed in Escherichia coli cells efficiently released SUMO-1 from SUMO-1. beta galactosidase fusion but not from other ubiquitin-like protein fusions, including Smt3.beta-galactosidase, suggesting its role in the generation of matured SUMO-1 specifically from its precursors. Interestingly, reproductive organs, such as testis, ovary, and prostate, contained much higher amounts of SUSP1 mRNA than colon and peripheral blood leukocyte, whereas other tissues, such as heart and spleen, had little or none. In addition, confocal microscopy using green fluorescent protein.SUSP1 fusion showed that SUSP1 is exclusively localized to the cytoplasm of NIH3T3 and HeLa cells. These results suggest that SUSP1 may play a role in the regulation of SUMO-1-mediated cellular processes particularly related to reproduction. PMID- 10799486 TI - Rb protein down-regulates the stress-activated signals through inhibiting c-Jun N terminal kinase/stress-activated protein kinase. AB - The Rb protein is the product of the retinoblastoma susceptibility gene and loss of Rb function is detected in many types of human cancers. Rb plays important roles in the regulation of cell proliferation, differentiation, senescence, and apoptotic cell death. Here we show that Rb can physically interact with c-Jun NH(2)-terminal kinase/stress-activated protein kinase (JNK/SAPK), thereby inhibiting intracellular signals mediated by JNK/SAPK. Both in vitro binding and in vitro kinase studies suggest that a carboxyl-terminal domain of Rb containing amino acids 768-928 might be crucial for inhibiting JNK/SAPK. In comparison, Rb did not affect enzymatic activity of either extracellular signal-regulated kinase 1 or p38. Ectopically expressed Rb also abrogated the apoptotic cell death induced by ultraviolet radiation or the activation of MEKK1, an upstream kinase that can stimulate the JNK/SAPK cascade. JNK/SAPK inhibition highlights a novel function of Rb, which may provide a new mechanism by which Rb regulates cell death. JNK/SAPK is a major protein kinase that can be stimulated in response to a variety of cellular stresses. Our results, therefore, suggest that Rb, by inhibiting JNK/SAPK, may act as a negative regulator in stress-activated intracellular signaling cascades. PMID- 10799488 TI - Primary structure requirements for Xenopus nodal-related 3 and a comparison with regions required by Xenopus nodal-related 2. AB - Transforming growth factor-beta superfamily members play important roles in the early development of animals. Activin and the Xenopus nodal related proteins 1, 2, and 4 induce muscle actin from Xenopus ectodermal explants, whereas the bone morphogenetic proteins 4 and 7 induce ectoderm to differentiate as epidermis. Bone morphogenetic proteins are antagonized by soluble binding proteins such as noggin and chordin, which leads to expression of neural cell adhesion molecule in animal caps. The transforming growth factor-beta superfamily member Xenopus nodal related 3 also induces the neural cell adhesion molecule through inhibition of bone morphogenetic proteins. Therefore, whereas Xenopus nodal-related 2 and 3 share a high amount of sequence homology, they lead to very different cell fates. This study investigates the functional domains that distinguish the activities of these two factors. It was found that mutually exclusive regions of nodal-related 2 and 3 were required for activity. The central region of the mature domain is required for nodal-related 2 to induce muscle actin, whereas the N- and C terminal ends of the mature domain are required for nodal-related 3 to induce neural cell adhesion molecule. These results help to define the minimal domains required for the unique activities of these factors. PMID- 10799487 TI - The active site of the thermophilic CYP119 from Sulfolobus solfataricus. AB - CYP119 from Sulfolobus solfataricus, the first thermophilic cytochrome P450, is stable at up to 85 degrees C. UV-visible and resonance Raman show the enzyme is in the low spin state and only modestly shifts to the high spin state at higher temperatures. Styrene only causes a small spin state shift, but T(1) NMR studies confirm that styrene is bound in the active site. CYP119 catalyzes the H(2)O(2) dependent epoxidation of styrene, cis-beta-methylstyrene, and cis-stilbene with retention of stereochemistry. This catalytic activity is stable to preincubation at 80 degrees C for 90 min. Site-specific mutagenesis shows that Thr-213 is catalytically important and Thr-214 helps to control the iron spin state. Topological analysis by reaction with aryldiazenes shows that Thr-213 lies above pyrrole rings A and B and is close to the iron atom, whereas Thr-214 is some distance away. CYP119 is very slowly reduced by putidaredoxin and putidaredoxin reductase, but these proteins support catalytic turnover of the Thr-214 mutants. Protein melting curves indicate that the thermal stability of CYP119 does not depend on the iron spin state or the active site architecture defined by the threonine residues. Independence of thermal stability from active site structural factors should facilitate the engineering of novel thermostable catalysts. PMID- 10799489 TI - Function and properties of chimeric MPR 46-MPR 300 mannose 6-phosphate receptors. AB - The two known mannose 6-phosphate receptors (MPR 46 and MPR 300) mediate the transport of mannose 6-phosphate-containing lysosomal proteins to lysosomes. Endocytosis of extracellular mannose 6-phosphate ligands can only be mediated by MPR 300. Neither type of MPR appears to be sufficient for targetting the full complement of lysosomal enzymes to lysosomes. The complements of lysosomal enzymes transported by either of the two receptors are distinct but largely overlapping. Chimeric receptors were constructed in which the transmembrane and cytoplasmic domains of the two receptors were systematically exchanged. After expression of the chimeric receptors in cells lacking endogenous MPRs the binding of ligands, the subcellular distribution and the sorting efficiency for lysosomal enzymes were analyzed. All chimeras were functional, and their subcellular distribution was similar to that of wild type MPRs. The ability to endocytose lysosomal enzymes was restricted to receptors with the lumenal domain of MPR 300. The efficiency to sort lysosomal enzymes correlated with the lumenal and cytoplasmic domains of MPR 300. In contrast to the wild type receptors, a significant fraction of most of the chimeric receptors was misrouted to lysosomes, indicating that the signals determining the routing of MPRs have been fitted for the parent receptor polypeptides. PMID- 10799490 TI - Hypoxic induction of prolyl 4-hydroxylase alpha (I) in cultured cells. AB - Accumulated evidence indicates that hypoxia activates collagen synthesis in tissues. To explore the molecular mechanism of activation, we screened genes that are up-regulated or down-regulated by hypoxia. Fibroblasts isolated from fetal rat lung were cultured under hypoxia. Differential display technique showed that the mRNA level of prolyl 4-hydroxylase (PH) alpha(I), an active subunit that catalyzes the oxygen-dependent hydroxylation of proline residue in procollagen, increased 2-3-fold after an 8-h exposure to hypoxia. This elevated level was maintained over 40 h and returned to the basal level after reoxygenation. The transcription rate, protein level, and hydroxyproline content (an indicator of the prolyl hydroxylation) were all elevated by hypoxic culture. Analysis of the promotor region of PHalpha(I) gene indicated that a motif similar to hypoxia responsive element (HRE) of hypoxia-inducible genes such as erythropoietin, was identified within a 120-base pair sequence upstream of the transcription start site. Luciferase reporter assay and mutational analysis showed that a site similar to the HRE in this motif is functionally essential to hypoxic response. Electrophoretic mobility shift assay revealed that hypoxia-inducible factor-1 was stimulated and bound to the PHalpha(I) HRE upon hypoxic challenge. Our results indicate that PHalpha(I), an essential enzyme for collagen synthesis, is a target gene for hypoxia-inducible factor-1. PMID- 10799491 TI - Glycosyl phosphatidylinositol myristoylation in African trypanosomes. New intermediates in the pathway for fatty acid remodeling. AB - Glycosyl phosphatidylinositol (GPI) anchors in the bloodstream form of Trypanosoma brucei are unusual in that their two fatty acids are myristate. The myristates are added in the final stages of GPI biosynthesis in a remodeling reaction. Remodeling occurs first at the sn-2 position of glycerol, involving removal of a longer fatty acid and subsequent attachment of myristate. The second myristate is then incorporated into the sn-1 position, but the mechanism has been unclear due to the unavailability of a reliable cell-free system supporting complete remodeling. Here, we first refined the cell-free system (by removing Mn(2+) ions), thereby allowing efficient production of the dimyristoylated GPI precursor. Using this improved system, we made three new discoveries concerning the pathway for fatty acid remodeling. First, we discovered a monomyristoylated GPI (known as glycolipid theta') as an intermediate involved in remodeling at the sn-1 position. Second, we found an alternative pathway for production of glycolipid theta, the first lyso intermediate in remodeling. The alternative pathway involves an inositol-acylated GPI known as glycolipid lyso-C'. Finally, we found that there is significant breakdown of GPIs during remodeling in the cell-free system, and we speculate that this breakdown has a regulatory role in GPI biosynthesis. PMID- 10799492 TI - Glucose-regulated turnover of mRNA and the influence of poly(A) tail length on half-life. AB - Glucose repression in Saccharomyces cerevisiae can now be seen to operate at two levels: regulation of transcription of certain genes and control of the half-life of the corresponding mRNAs (Scheffler, I. E., de la Cruz, B. J., and Prieto, S. (1998) Int. J. Biochem Cell Biol. 30, 1175-1193). For example, the steady state levels of SDH2 mRNA and SUC2 mRNA are significantly determined by their differential rates of turnover. A current model for the mechanism of mRNA turnover includes three distinct steps: a rate-limiting deadenylation, removal of the 5'-7-methyl-G (decapping), and 5'-3' exonuclease digestion. We have investigated the same three reactions during glucose-induced degradation of these transcripts. Our results indicate that while decapping (by Dcp1p) and 5'-3' exonuclease digestion (by Xrn1p) are obligatory steps for the rapid degradation of these mRNAs, the dependence on deadenylation is more complicated. At steady state in glycerol these transcripts have very short poly(A) tails but are nevertheless very stable; the addition of glucose causes immediate decapping and degradation without further deadenylation; in contrast, newly made SUC2 mRNA (after a shift from glucose to glycerol) has significantly longer poly(A) tails, and such transcripts are not rapidly degraded upon addition of glucose. A constitutive deadenylation reaction that is independent of the carbon source eventually makes the stability of these transcripts very sensitive to glucose. These results are interpreted in terms of a working hypothesis proposing a competition between translational initiation and decapping influenced by the carbon source. The presence of a long poly(A) tail may also affect this competition in favor of translational initiation and mRNA stabilization. PMID- 10799493 TI - Identification and characterization of a putative active site for peptide methionine sulfoxide reductase (MsrA) and its substrate stereospecificity. AB - Peptide methionine sulfoxide reductases (MsrA) from many different organisms share a consensus amino acid sequence (GCFWG) that could play an important role in their active site. Site-directed single substitution of each of these amino acids except glycines in the yeast MsrA resulted in total loss of enzyme activity. Nevertheless, all the recombinant MsrA mutants and native proteins had a very similar circular dichroism spectrum. The demonstration that either treatment with iodoacetamide or replacement of the motif cysteine with serine leads to inactivation of the enzyme underscores the singular importance of cysteine residues in the activity of MsrA. The recombinant yeast MsrA was used for general characterization of the enzyme. Its K(m) value was similar to the bovine MsrA and appreciably lower than the K(m) of the bacterial enzyme. Also, it was shown that the enzymatic activity increased dramatically with increasing ionic strength. The recombinant yeast MsrA activity and the reduction activity of free methionine sulfoxide(s) were stereoselective toward the L-methionine S sulfoxide and S-methyl p-tolyl sulfoxide. It was established that a methionine auxotroph yeast strain could grow on either form of L-methionine sulfoxide. PMID- 10799494 TI - Divergent signaling pathways requiring discrete calcium signals mediate concurrent activation of two mitogen-activated protein kinases by gonadotropin releasing hormone. AB - Receptors coupled to heterotrimeric G proteins are linked to activation of mitogen-activated protein kinases (MAPKs) via receptor- and cell-specific mechanisms. We have demonstrated recently that gonadotropin-releasing hormone (GnRH) receptor occupancy results in activation of extracellular signal-regulated kinase (ERK) through a mechanism requiring calcium influx through L-type calcium channels in alphaT3-1 cells and primary rat gonadotropes. Further studies were undertaken to explore the signaling mechanisms by which the GnRH receptor is coupled to activation of another member of the MAPK family, c-Jun N-terminal kinase (JNK). GnRH induces activation of the JNK cascade in a dose-, time-, and receptor-dependent manner in clonal alphaT3-1 cells and primary rat pituitary gonadotrophs. Coexpression of dominant negative Cdc42 and kinase-defective p21 activated kinase 1 and MAPK kinase 7 with JNK and ERK indicated that specific activation of JNK by GnRH appears to involve these signaling molecules. Unlike ERK activation, GnRH-stimulated JNK activity does not require activation of protein kinase C and is not blocked after chelation of extracellular calcium with EGTA. GnRH-induced JNK activity was reduced after treatment with the intracellular calcium chelator BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid acetoxymethyl ester), whereas activation of ERK was not affected. Chelation of intracellular calcium also reduced GnRH-induced activation of JNK in rat pituitary cells in primary culture. GnRH-induced induction and activation of the JNK target c-Jun was inhibited after chelation of intracellular calcium, whereas induction of c-Fos, a known target of ERK, was unaffected. Therefore, although activation of ERK by GnRH requires a specific influx of calcium through L-type calcium channels, JNK activation is independent of extracellular calcium but sensitive to chelation of intracellular calcium. Our results provide novel evidence that GnRH activates two MAPK superfamily members via strikingly divergent signaling pathways with differential sensitivity to activation of protein kinase C and mobilization of discrete pools of calcium. PMID- 10799495 TI - Downstream promoter sequences facilitate the formation of a specific transcription factor IID-promoter complex topology required for efficient transcription from the megalin/low density lipoprotein receptor-related protein 2 promoter. AB - Megalin/low density lipoprotein receptor-related protein 2 (LRP-2) is an endocytic receptor expressed in highly specialized cell types such as parathyroid cells and epithelia of the kidney. Previous experiments identified a nonconsensus TATA element, with the sequence TAGAAAA, as crucial for accurate and efficient transcription from the LRP-2 promoter. Here we show that, in addition to the TAGA element, promoter sequences downstream of the transcription start site contribute significantly to transcription both in vitro and in transfected cells. Deletion and point mutational analyses reveal that the promoter region located between positions +5 and +11 (sequence TTTTGGC) is of particular importance. Complementation experiments in nuclear extracts lacking transcription factor IID (TFIID) activity show that TATA-binding protein-associated factors of TFIID are essential for the function of LRP-2 downstream promoter sequences. Interestingly, DNase I footprinting studies show that the downstream region between positions +5 and +11 does not significantly affect overall TFIID affinity to the promoter but that it profoundly affects the topology of the TFIID x promoter complex not only downstream of the transcription start site, but in particular in the TATA box region. Our observations suggest a model for a novel downstream sequence function, in which TATA-binding protein-associated factor-promoter interactions downstream of the transcription start site modulate TFIID-DNA interactions in the TATA box region. PMID- 10799496 TI - Tyrosine 62 of the gamma-aminobutyric acid type A receptor beta 2 subunit is an important determinant of high affinity agonist binding. AB - The gamma-aminobutyric acid type A receptor (GABA(A)R) carries both high (K(D) = 10-30 nm) and low (K(D) = 0.1-1.0 microm) affinity binding sites for agonists. We have used site-directed mutagenesis to identify a specific residue in the rat beta2 subunit that is involved in high affinity agonist binding. Tyrosine residues at positions 62 and 74 were mutated to either phenylalanine or serine and the effects on ligand binding and ion channel activation were investigated after the expression of mutant subunits with wild-type alpha1 and gamma2 subunits in tsA201 cells or in Xenopus oocytes. None of the mutations affected [(3)H]Ro15 4513 binding or impaired allosteric interactions between the low affinity GABA and benzodiazepine sites. Although mutations at position 74 had little effect on [(3)H]muscimol binding, the Y62F mutation decreased the affinity of the high affinity [(3)H]muscimol binding sites by approximately 6-fold, and the Y62S mutation led to a loss of detectable high affinity binding sites. After expression in oocytes, the EC(50) values for both muscimol and GABA-induced activation of Y62F and Y62S receptors were increased by 2- and 6-fold compared with the wild-type. We conclude that Tyr-62 of the beta subunit is an important determinant for high affinity agonist binding to the GABA(A) receptor. PMID- 10799497 TI - The unfolding pathway and conformational stability of potato carboxypeptidase inhibitor. AB - The unfolding and denaturation curves of potato carboxypeptidase inhibitor (PCI) were investigated using the technique of disulfide scrambling. In the presence of denaturant and thiol initiator, the native PCI denatures by shuffling its native disulfide bonds and converts to form a mixture of scrambled PCI that consists of 9 out of a possible 14 isomers. The denaturation curve is determined by the fraction of native PCI converted to scrambled isomers under increasing concentrations of denaturant. The concentration of guanidine thiocyanate, guanidine hydrochloride, and urea required to denature 50% of the native PCI was found to be 0.7, 1.45, and 8 m, respectively. The PCI unfolding curve was constructed through the analysis of structures of scrambled isomers that were denatured under increasing concentrations of denaturant. These results reveal the existence of structurally defined unfolding intermediates and a progressive expansion of the polypeptide chain. The yield of the beads-form isomer (Cys(8) Cys(12), Cys(18)-Cys(24), and Cys(27)-Cys(34)) as a fraction of total denatured PCI was shown to be directly proportional to the strength of the denaturing condition. Furthermore, the PCI sequence was unable to fold quantitatively into a single native structure. Under physiological conditions, the scrambled isomers of PCI that constitute about 4% of the protein were in equilibrium with native PCI. PMID- 10799498 TI - Identification of a novel isopeptidase with dual specificity for ubiquitin- and NEDD8-conjugated proteins. AB - Covalent conjugation of proteins by ubiquitin or ubiquitin-like molecules is an important form of post-translational modification and plays a critical role in many cellular processes. Similar to the concept of phosphorylation and dephosphorylation, these conjugates are regulated by a large number of deconjugating enzymes. Here, we report the cloning of a 2,141-base pair DNA fragment from human placenta cDNA library by a strategy that involves expressed sequence tag data base searching, polymerase chain reaction, and rapid amplification of cDNA ends. Nucleotide sequence analysis revealed that the cloned cDNA contains an open reading frame of 1,143 base pairs encoding a novel protease, USP21, which is composed of 381 residues with a calculated molecular mass of 43 kDa. The human USP21 gene is located on chromosome 1q21 and encodes a member of the ubiquitin-specific protease family with highly conserved Cys and His domains. The activity and specificity of USP21 were determined by using a COS cell expression system in vivo. We showed that USP21 is capable of removing ubiquitin from ubiquitinated proteins as expected. Furthermore, USP21 is capable of removing NEDD8 from NEDD8 conjugates but has no effect on Sentrin-1 conjugates. As expected from its biochemical activity, overexpression of USP21 has a profound growth inhibitory effect on U2OS cells. Thus, USP21 is the first ubiquitin-specific protease shown to have dual specificity for both ubiquitin and NEDD8 and may play an important role in the regulation of cell growth. PMID- 10799499 TI - Complementation of a glucose transporter mutant of Schizosaccharomyces pombe by a novel Trypanosoma brucei gene. AB - The African trypanosome Trypanosoma brucei has a digenetic life cycle that involves the insect vector and the mammalian host. This is underscored by biochemical switches in its nutritional requirements. In the insect vector, the parasite relies on amino acid catabolism, but in the mammalian host, it derives its energy exclusively from blood glucose. Glucose transport is facilitated, and constitutes the rate-limiting step in ATP synthesis. Here, we report the cloning of a novel glucose transporter-related gene by heterologous screening of a lambdaEMBL4 genomic library of T. brucei EATRO 164 using a rat liver glucose transporter cDNA clone. Genomic analysis shows that the gene is present as a single copy within the parasite genome. The gene encodes a protein with an estimated molecular mass of 55.9 kDa, which shares only segmental homology with members of the glucose transporter superfamily. Several potential post translational modification sites including phosphorylation, N-glycosylation, and cotranslational myristoylation sites also punctuate the sequence. It is distinguished from classical transporter proteins by the absence of putative hydrophobic membrane-spanning domains. However, this protein was capable of complementing Schizosaccharomyces pombe glucose transporter mutants. The rescued phenotype conferred the ability of the cells to grow on a broad range of sugars, both monosaccharides and disaccharides. The kinetics of glucose uptake reflected those in T. brucei. In addition to complementation in yeast, we also showed that the gene enhanced glucose uptake in cultured mammalian cells. PMID- 10799500 TI - Binding of multivalent carbohydrates to concanavalin A and Dioclea grandiflora lectin. Thermodynamic analysis of the "multivalency effect". AB - Binding of a series of synthetic multivalent carbohydrate analogs to the Man/Glc specific lectins concanavalin A and Dioclea grandiflora lectin was investigated by isothermal titration microcalorimetry. Dimeric analogs possessing terminal alpha-D-mannopyranoside residues, and di-, tri-, and tetrameric analogs possessing terminal 3, 6-di-O-(alpha-D-mannopyranosyl)-alpha-D-mannopyranoside residues, which is the core trimannoside of asparagine-linked carbohydrates, were selected in order to compare the effects of low and high affinity analogs, respectively. Experimental conditions were found that prevented precipitation of the carbohydrate-lectin cross-linked complexes during the isothermal titration microcalorimetry experiments. The results show that the value of n, the number of binding sites on each monomer of the lectins, is inversely proportional to the number of binding epitopes (valency) of each carbohydrate. Hence, n values close to 1.0, 0.50, and 0.25 were observed for the binding of mono-, di-, and tetravalent sugars, respectively, to the two lectins. Importantly, differences in the functional valency of a triantennary analog for concanavalin A and D. grandiflora lectin are observed. The enthalpy of binding, DeltaH, is observed to be directly proportional to the number of binding epitopes in the higher affinity analogs. For example, DeltaH of a tetravalent trimannoside analog is nearly four times greater than that of the corresponding monovalent analog. Increases in K(a) values of the multivalent carbohydrates relative to monovalent analogs, known as the "multivalency effect," are shown to be due to more positive entropy (TDeltaS) contributions to binding of the former sugars. A general thermodynamic model for distinguishing binding of multivalent ligands to a single receptor with multiple, equal subsites versus binding to separate receptor molecules is given. PMID- 10799501 TI - Cdc42-induced activation of the mixed-lineage kinase SPRK in vivo. Requirement of the Cdc42/Rac interactive binding motif and changes in phosphorylation. AB - Src homology 3 domain (SH3)-containing proline-rich protein kinase (SPRK)/mixed lineage kinase (MLK)-3 is a serine/threonine kinase that upon overexpression in mammalian cells activates the c-Jun NH(2)-terminal kinase pathway. The mechanisms by which SPRK activity is regulated are not well understood. The small Rho family GTPases, Rac and Cdc42, have been shown to bind and modulate the activities of signaling proteins, including SPRK, which contain Cdc42/Rac interactive binding motifs. Coexpression of SPRK and activated Cdc42 increases SPRKs activity. SPRKs Cdc42/Rac interactive binding-like motif contains six of the eight consensus residues. Using a site-directed mutagenesis approach, we show that SPRK contains a functional Cdc42/Rac interactive binding motif that is required for SPRKs association with and activation by Cdc42. However, experiments using a SPRK variant that lacks the COOH-terminal zipper region/basic stretch suggest that this region may also contribute to Cdc42 binding. Unlike the PAK family of protein kinases, we find that the activation of SPRK by Cdc42 cannot be recapitulated in an in vitro system using purified, recombinant proteins. Comparative phosphopeptide mapping demonstrates that coexpression of activated Cdc42 with SPRK alters the in vivo serine/threonine phosphorylation pattern of SPRK suggesting that the mechanism by which Cdc42 increases SPRKs catalytic activity involves a change in the in vivo phosphorylation of SPRK. This is, to the best of our knowledge, the first demonstrated example of a Cdc42-mediated change in the in vivo phosphorylation of a protein kinase. These studies suggest an additional component or cellular environment is required for SPRK activation by Cdc42. PMID- 10799502 TI - Ornithine decarboxylase of Stagonospora (Septoria) nodorum is required for virulence toward wheat. AB - A knockout strain of Stagonospora (Septoria) nodorum lacking the single ornithine decarboxylase (ODC) allele has been created by targeted gene replacement. A central region of the S. nodorum ODC gene was isolated by polymerase chain reaction using degenerate oligonucleotides and used to probe a lambda genomic library. The gene was sequenced and the encoded ODC protein sequence was shown to be similar to those from other fungi. The functionality of the S. nodorum ODC was confirmed by complementation of an Aspergillus nidulans mutant (puA) strain devoid of ODC activity, restoring growth in the absence of exogenous polyamines. Sporulation of the transformants was reduced suggesting abberant regulation of the S. nodorum gene in A. nidulans. Transformation-mediated gene replacement was used to create strains which were auxotrophic for putrescine and lack ODC coding sequences. Pathogenicity studies on these mutants showed that they are greatly reduced in virulence compared with non-disrupted transformants. This confirms that the strains carrying an ODC disruption cannot obtain sufficient polyamines from the host plant for normal growth and, thus, that fungal ODC may be a suitable target for chemical intervention. PMID- 10799503 TI - The prodomain of caspase-1 enhances Fas-mediated apoptosis through facilitation of caspase-8 activation. AB - Caspase-1 (interleukin-1beta converting enzyme) is produced in the form of a latent precursor, which is cleaved to yield a prodomain in addition to the p20 and p10 subunits. It has been established that the (p20/p10)(2) heterotetramer processes the latent precursor of interleukin-1beta into an active form during apoptosis, but the function of the residual prodomain of caspase-1 (Pro-C1) has not been established. To evaluate the involvement of Pro-C1 in apoptosis, a Pro C1 expression vector was transfected into the HeLa cell line, which is susceptible to Fas-mediated apoptosis. Expression of recombinant Pro-C1 in HeLa cells enhanced apoptosis mediated by Fas, but not etoposide-induced apoptosis. This enhancement of Fas-mediated apoptosis was abolished by inhibitors of caspase 8 (Ile-Glu-Thr-Asp-fluoromethyl ketone) and caspase-3 (Asp-Glu-Val-Asp-aldehyde) but was only slightly diminished by an inhibitor of caspase-1 (acetyl-Tyr-Val-Ala Asp-chloromethyl ketone). During apoptosis induced by an agonistic anti-Fas antibody, the activation of caspase-8 and caspase-3 was more pronounced and occurred more rapidly in HeLa/Pro-C1 cells than in the empty vector transfectant (HeLa/vec) cells; in contrast, caspase-1 was not activated in either HeLa/Pro-C1 or HeLa/vec cells. These results demonstrate an additional and novel function for caspase-1 in which Pro-C1 acts to enhance Fas-mediated apoptosis, most probably through facilitation of the activation of caspase-8. PMID- 10799504 TI - The role of the DELSEED motif of the beta subunit in rotation of F1-ATPase. AB - F(1)-ATPase is a rotary motor protein, and ATP hydrolysis generates torque at the interface between the gamma subunit, a rotor shaft, and the alpha(3)beta(3) substructure, a stator ring. The region of conserved acidic "DELSEED" motif of the beta subunit has a contact with gamma subunit and has been assumed to be involved in torque generation. Using the thermophilic alpha(3)beta(3)gamma complex in which the corresponding sequence is DELSDED, we replaced each residue and all five acidic residues in this sequence with alanine. In addition, each of two conserved residues at the counterpart contact position of gamma subunit was also replaced. Surprisingly, all of these mutants rotated with as much torque as the wild-type. We conclude that side chains of the DELSEED motif of the beta subunit do not have a direct role in torque generation. PMID- 10799505 TI - The human nm23-H4 gene product is a mitochondrial nucleoside diphosphate kinase. AB - We demonstrate here the catalytic activity and subcellular localization of the Nm23-H4 protein, product of nm23-H4, a new member of the human nm23/nucleoside diphosphate (NDP) kinase gene family (Milon, L., Rousseau-Merck, M., Munier, A., Erent, M., Lascu, I., Capeau, J., and Lacombe, M. L. (1997) Hum. Genet. 99, 550 557). Nm3-H4 was synthesized in escherichia coli as the full-length protein and as a truncated form missing the N-terminal extension characteristic of mitochondrial targeting. The truncated form possesses NDP kinase activity, whereas the full-length protein is inactive, suggesting that the extension prevents enzyme folding and/or activity. X-ray crystallographic analysis was performed on active truncated Nm23-H4. Like other eukaryotic NDP kinases, it is a hexamer. Nm23-H4 naturally possesses a serine residue at position 129, equivalent to the K-pn mutation of the Drosophila NDP kinase. The x-ray structure shows that the presence of Ser(129) has local structural effects that weaken subunit interactions. Site-directed mutagenesis shows that the serine is responsible for the lability of Nm23-H4 to heat and urea treatment, because the S129P mutant is greatly stabilized. Examination of human embryonic kidney 293 cells transfected with green fluorescent protein fusions by confocal microscopy shows a specific mitochondrial localization of Nm23-H4 that was also demonstrated by Western blot analysis of subcellular fractions of these cells. Import into mitochondria is accompanied by cleavage of the N-terminal extension that results in NDP kinase activity. Submitochondrial fractionation indicates that Nm23-H4 is associated with mitochondrial membranes, possibly to the contact sites between the outer and inner membranes. PMID- 10799506 TI - Characterization of the heparin-binding site of the mycobacterial heparin-binding hemagglutinin adhesin. AB - The mycobacterial adhesin heparin-binding hemagglutinin (HBHA) contains several lysine-rich repeats at its carboxyl-terminal end. Using truncated recombinant HBHA forms and hybrid proteins containing HBHA repeats grafted onto the Escherichia coli maltose-binding protein (MBP), we found that these repeats are responsible for heparin binding. Immunofluorescence microscopy studies revealed that their deletion abrogates binding of HBHA to human pneumocytes. Conversely, when fused to MBP, the HBHA repeats confer pneumocyte adherence properties to the hybrid protein. Treatment of pneumocytes with glycosaminoglycan-degrading enzymes showed that HBHA binding depends on the presence of heparan sulfate chains on the cell surface. The epitope of a monoclonal antibody that inhibits mycobacterial adherence to epithelial cells was mapped within the lysine-rich repeats, confirming their involvement in mycobacterial adherence to epithelial cells. Surface plasmon resonance analyses showed that recombinant HBHA binds to immobilized heparin with fast association kinetics (k(a) = 5.62 (+/- 0.10) x 10(5) m(-1) s(-1)), whereas the dissociation kinetics were slower (k(d) = 0.015 (+/- 0.002) s(-1)), yielding a K(D) value of 26 nm. Similar analyses with grafted MBP indicated similar kinetic constants, indicating that the carboxyl-terminal repeats contain the entire heparin-binding site of HBHA. The molecular characterization of the interactions of HBHA with epithelial glycosaminoglycans should help to better understand mycobacterial adherence within the lungs and may ultimately lead to new approaches for therapy or immunoprophylaxis. PMID- 10799507 TI - Characterization of a novel sphingosine 1-phosphate receptor, Edg-8. AB - Three G protein-coupled receptors (Edg-1, Edg-3, and Edg-5) for the lysolipid phosphoric acid mediator sphingosine 1-phosphate have been described by molecular cloning. Using a similar sequence that we found in the expressed sequence tag data base, we cloned and characterized of a fourth, high affinity, rat brain sphingosine 1-phosphate receptor, Edg-8. When HEK293T cells were co-transfected with Edg-8 and G protein DNAs, prepared membranes showed sphingosine 1- phosphate dependent increases in [(35)S]guanosine 5'-(3-O-thio)triphosphate binding with an EC(50) of 90 nm. In a rat hepatoma Rh7777 cell line that exhibits modest endogenous responses to sphingosine 1-phosphate, this lipid mediator inhibited forskolin-driven rises in cAMP by greater than 90% when the cells were transfected with Edg-8 DNA (IC(50) 0.7 nm). This response is blocked fully by prior treatment of cultures with pertussis toxin, thus implicating signaling through G(i/o)alpha proteins. Furthermore, Xenopus oocytes exhibit a calcium response to sphingosine 1-phosphate after injection of Edg-8 mRNA, but only when oocytes are co-injected with chimeric G(q/i)alpha protein mRNA. Membranes from HEK293T and Rh7777 cell cultures expressing Edg-8 exhibited high affinity (K(D) = 2 nm) binding for radiolabeled sphingosine 1-phosphate. Rat Edg-8 RNA is expressed in spleen and throughout adult rat brain where in situ hybridization revealed it to be associated with white matter. Together our data demonstrate that Edg-8 is a high affinity sphingosine 1-phosphate receptor that couples to G(i/o)alpha proteins and is expressed predominantly by oligodendrocytes and/or fibrous astrocytes in the rat brain. PMID- 10799508 TI - Development of the structural basis for antimicrobial and hemolytic activities of peptides based on gramicidin S and design of novel analogs using NMR spectroscopy. AB - The structures of 14-residue head-to-tail cyclic gramicidin S peptides have been investigated to develop the structural rationale for their antimicrobial and hemolytic profiles. The basis for these studies is GS14 (cyclo(VKLKVdYPLKVKLdYP)), designed as an extension of the naturally occurring antimicrobial peptide. The structure of GS14 has been determined using NMR methods and was found to exist in a highly amphipathic antiparallel beta-sheet conformation. Systematic enantiomeric substitutions within the framework of the GS14 peptide were found to decrease the amphipathicity of this molecule. These results indicated that there was a direct correlation between the high amphipathic character and potent hemolytic activity in the diastereomers, whereas an inverse correlation existed between amphipathicity and antimicrobial function. To define the structural consequences of changing the amphipathic nature of GS14 analogs to maximize antimicrobial activity and to minimize hemolysis, NMR structures were determined in water and the membrane-mimetic solvent trifluoroethanol. The structures show that these attributes are the result of induction of the beta-sheet character in a membrane environment and the positioning of charged side chains on the hydrophobic face of the cyclic framework, thus decreasing the amphipathicity and directed hydrophobicity of these molecules. Implications for the design of more effective antimicrobials are discussed. PMID- 10799509 TI - Identification and characterization of CKIP-1, a novel pleckstrin homology domain containing protein that interacts with protein kinase CK2. AB - The catalytic subunits of protein kinase CK2, CK2alpha and CK2alpha', are closely related to each other but exhibit functional specialization. To test the hypothesis that specific functions of CK2alpha and CK2alpha' are mediated by specific interaction partners, we used the yeast two-hybrid system to identify CK2alpha- or CK2alpha'-binding proteins. We report the identification and characterization of a novel CK2-interacting protein, designated CKIP-1, that interacts with CK2alpha, but not CK2alpha', in the yeast two-hybrid system. CKIP 1 also interacts with CK2alpha in vitro and is co-immunoprecipitated from cell extracts with epitope-tagged CK2alpha and an enhanced green fluorescent protein fusion protein encoding CKIP-1 (i.e. EGFP-CKIP-1) when they are co-expressed. CK2 activity is detected in anti-CKIP-1 immunoprecipitates performed with extracts from non-transfected cells indicating that CKIP-1 and CK2 interact under physiological conditions. The CKIP-1 cDNA is broadly expressed and encodes a protein with a predicted molecular weight of 46,000. EGFP-CKIP-1 is localized within the nucleus and at the plasma membrane. The plasma membrane localization is dependent on the presence of an amino-terminal pleckstrin homology domain. We postulate that CKIP-1 is a non-enzymatic regulator of one isoform of CK2 (i.e. CK2alpha) with a potential role in targeting CK2alpha to a particular cellular location. PMID- 10799510 TI - The lymphotoxin-beta receptor is necessary and sufficient for LIGHT-mediated apoptosis of tumor cells. AB - LIGHT is a tumor necrosis factor (TNF) ligand superfamily member, which binds two known cellular receptors, lymphotoxin-beta receptor (LTbetaR) and the herpesvirus entry mediator (HveA). LIGHT is a homotrimer that activates proapoptotic and integrin-inducing pathways. Receptor binding residues via LIGHT were identified by introducing point mutations in the A' --> A" and D --> E loops of LIGHT, which altered binding to LTbetaR and HveA. One mutant of LIGHT exhibits selective binding to HveA and is inactive triggering cell death in HT29.14s cells or induction of ICAM-1 in fibroblasts. Studies with HveA- or LTbetaR-specific antibodies further indicated that HveA does not contribute, either cooperatively or by direct signaling, to the death pathway activated by LIGHT. LTbetaR, not HveA, recruits TNF receptor-associated factor-3 (TRAF3), and LIGHT-induced death is blocked by a dominant negative TRAF3 mutant. Together, these results indicate that TRAF3 recruitment propagates death signals initiated by LIGHT-LTbetaR interaction and implicates a distinct biological role for LIGHT-HveA system. PMID- 10799511 TI - Binding of the second generation non-nucleoside inhibitor S-1153 to HIV-1 reverse transcriptase involves extensive main chain hydrogen bonding. AB - S-1153 (AG1549) is perhaps the most promising non-nucleoside inhibitor of HIV-1 reverse transcriptase currently under development as a potential anti-AIDS drug, because it has a favorable profile of resilience to many drug resistance mutations. We have determined the crystal structure of S-1153 in a complex with HIV-1 reverse transcriptase. The complex possesses some novel features, including an extensive network of hydrogen bonds involving the main chain of residues 101, 103, and 236 of the p66 reverse transcriptase subunit. Such interactions are unlikely to be disrupted by side chain mutations. The reverse transcriptase/S 1153 complex suggests different ways in which resilience to mutations in the non nucleoside inhibitors of reverse transcriptase binding site can be achieved. PMID- 10799512 TI - Identification of the residues in the extracellular region of KDR important for interaction with vascular endothelial growth factor and neutralizing anti-KDR antibodies. AB - The kinase domain receptor (KDR) of vascular endothelial growth factor (VEGF) is the main human receptor responsible for the angiogenic activity of VEGF. The extracellular region of KDR is comprised of seven immunoglobulin-like domains, of which the first three have been shown to be required for ligand binding. We have previously described antibodies directed against the extracellular region of KDR, including MAB383 and MAB664, which were shown to block the binding of VEGF to the receptor and to inhibit both VEGF-induced mitogenesis of human endothelial cells in vitro and tumor growth in vivo. Here we generated a series of KDR deletion mutants consisting of truncated extracellular regions and mapped out the domain(s) responsible for binding to VEGF and the neutralizing anti-KDR antibodies. All neutralizing antibodies were found to require domain 3 for efficient binding. Alanine-scanning mutagenesis of domain 3 identified two different sets of five residues, Ile(256), Asp(257), Glu(261), Leu(313), and Thr(315) and Tyr(262), Pro(263), Ser(264), Ser(265), and Lys(266), that were critical for binding to MAB383 and MAB664, respectively. Combination of alanine mutations affecting both MAB383 and MAB664 binding resulted in a variant that also lost binding to VEGF. These results suggest that the residues within this region of domain 3 are critical for VEGF binding. Our studies provide a basis for the mechanism of action of our anti-KDR antibodies and establish a functional foundation for the development of other classes of antagonists to the receptor. PMID- 10799513 TI - Differential influence of the 4F2 heavy chain and the protein related to b(0,+) amino acid transport on substrate affinity of the heteromeric b(0,+) amino acid transporter. AB - We provide evidence here that b(0,+) amino acid transporter (b(0, +)AT) interacts with 4F2 heavy chain (4F2hc) as well as with the protein related to b(0,+) amino acid transporter (rBAT) to constitute functionally competent b(0,+)-like amino acid transport systems. This evidence has been obtained by co-expression of b(0, +)AT and 4F2hc or b(0,+)AT and rBAT in human retinal pigment epithelial cells and in COS-1 cells. The ability to interact with 4F2hc and rBAT is demonstrable with mouse b(0,+)AT as well as with human b(0,+)AT. Even though both the 4F2hc x b(0,+)AT complex and the rBAT x b(0,+)AT complex exhibit substrate specificity that is characteristic of system b(0,+), these two complexes differ significantly in substrate affinity. The 4F2hc x b(0,+)AT complex has higher substrate affinity than the rBAT x b(0,+)AT complex. In situ hybridization studies demonstrate that the regional distribution pattern of mRNA in the kidney is identical for b(0,+)AT and 4F2hc. The pattern of rBAT mRNA expression is different from that of b(0,+)AT mRNA and 4F2hc mRNA, but there are regions in the kidney where b(0,+)AT mRNA expression overlaps with rBAT mRNA expression as well as with 4F2hc mRNA expression. PMID- 10799514 TI - Hybrid proteasomes. Induction by interferon-gamma and contribution to ATP dependent proteolysis. AB - Eukaryotic cells contain various types of proteasomes. Core 20 S proteasomes (abbreviated 20 S below) have two binding sites for the regulatory particles, PA700 and PA28. PA700-20 S-PA700 complexes are known as 26 S proteasomes and are ATP-dependent machines that degrade cell proteins. PA28 is found both in previously described complexes of the type PA28-20 S-PA28 and in complexes that also contain PA700, as PA700-20 S-PA28. We refer to the latter as "hybrid proteasomes." The relative amounts of the various types of proteasomes in HeLa extracts were determined by a combination of immunoprecipitation and immunoblotting. Hybrid proteasomes accounted for about a fourth of all proteasomes in the extracts. Association of PA28 and proteasomes proved to be ATP dependent. Hybrid proteasomes catalyzed ATP-dependent degradation of ornithine decarboxylase (ODC) without ubiquitinylation, as do 26 S proteasomes. In contrast, the homo-PA28 complex (PA28-20 S-PA28) was incapable of degrading ODC. Intriguingly, a major immunomodulatory cytokine, interferon-gamma, appreciably enhanced the ODC degradation in HeLa and SW620 cells through induction of the hybrid proteasome, which may also be responsible for the immunological processing of intracellular antigens. Taken together, we report here for the first time the existence of two types of ATP-dependent proteases, the 26 S proteasome and the hybrid proteasome, which appear to share the ATP-dependent proteolytic pathway in mammalian cells. PMID- 10799515 TI - The Toxoplasma adhesive protein MIC2 is proteolytically processed at multiple sites by two parasite-derived proteases. AB - MIC2 is an adhesive protein that participates in host cell invasion by the obligate intracellular parasite Toxoplasma gondii. Earlier studies established that MIC2 is secreted into the culture medium by extracellular parasites and that release is coincident with proteolytic modification. Since little is known about proteolytic processing of proteins secreted by T. gondii, we undertook this study to investigate the proteolytic events that accompany secretion of MIC2. We demonstrate that the C-terminal domain of MIC2 is removed by a protease, termed MPP1, when MIC2 is released into the culture supernatant. Additionally, prior to release, a second protease, termed MPP2, trims the N terminus of MIC2, resulting in the release of heterogeneously sized species of MIC2. Although MPP1 activity was unaffected by any of the protease inhibitors tested, MPP2 activity was blocked by a subset of serine and cysteine protease inhibitors. These results establish that MIC2 is proteolytically modified at multiple sites by two distinct enzymes that probably operate on the parasite surface. PMID- 10799516 TI - Three-dimensional reconstructions of calcium/calmodulin-dependent (CaM) kinase IIalpha and truncated CaM kinase IIalpha reveal a unique organization for its structural core and functional domains. AB - Studies of the structural organization of calcium/ calmodulin-dependent protein kinase IIalpha (CaM KIIalpha) and truncated CaM KIIalpha by three-dimensional electron microscopy and protein engineering show that the structures consist of 12 subunits that are organized in two stacked hexameric rings with 622 symmetry. The body of CaM KIIalpha is gear-shaped, consisting of six slanted flanges, and has six foot-like processes attached by narrow appendages to both ends of the flanges. Truncated CaM KIIalpha that lacks functional domains has a structure that is very similar to the body of CaM KIIalpha. Thus, the functional domains reside in the foot-like processes, and the association domain comprises the gear shaped core. The ribbon diagram of the bilobate structure of CaM KI fits nicely in the envelope of the foot-like component and indicates that the crevice between the two lobes comprising the functional domains is near the middle portion of the foot. The clustering of the functional domains provides a favorable arrangement for the autophosphorylation reaction, and the unusual arrangement of the catalytic domain on extended tethers appears to be significant for the remarkable functional diversity of CaM KIIalpha in cellular regulation. PMID- 10799517 TI - Protein kinase A associates with cystic fibrosis transmembrane conductance regulator via an interaction with ezrin. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is an epithelial Cl(-) channel whose activity is controlled by cAMP-dependent protein kinase (PKA) mediated phosphorylation. We found that CFTR immunoprecipitates from Calu-3 airway cells contain endogenous PKA, which is capable of phosphorylating CFTR. This phosphorylation is stimulated by cAMP and inhibited by the PKA inhibitory peptide. The endogenous PKA that co-precipitates with CFTR could also phosphorylate the PKA substrate peptide, Leu-Arg-Arg-Ala-Ser-Leu-Gly (kemptide). Both the catalytic and type II regulatory subunits of PKA are identified by immunoblotting CFTR immunoprecipitates, demonstrating that the endogenous kinase associated with CFTR is PKA, type II (PKA II). Phosphorylation reactions mediated by CFTR-associated PKA II are inhibited by Ht31 peptide but not by the control peptide Ht31P, indicating that a protein kinase A anchoring protein (AKAP) is responsible for the association between PKA and CFTR. Ezrin may function as this AKAP, since it is expressed in Calu-3 and T84 epithelia, ezrin binds RII in overlay assays, and RII is immunoprecipitated with ezrin from Calu-3 cells. Whole cell patch clamp of Calu-3 cells shows that Ht31 peptide reduces cAMP-stimulated CFTR Cl(-) current, but Ht31P does not. Taken together, these data demonstrate that PKA II is linked physically and functionally to CFTR by an AKAP interaction, and they suggest that ezrin serves as an AKAP for PKA-mediated phosphorylation of CFTR. PMID- 10799518 TI - Regulation of phosphatidylcholine metabolism in Chinese hamster ovary cells by the sterol regulatory element-binding protein (SREBP)/SREBP cleavage-activating protein pathway. AB - Sterol regulation-defective (SRD) 4 cells expressing a mutant sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP D443N) and Chinese hamster ovary (CHO) cells stably expressing SCAP (CHO-SCAP) and SCAP D443N (CHO-SCAP-D443N) have increased cholesterol and fatty acid synthesis because of constitutive processing of SREBPs. We assessed whether constitutive activation of SREBPs also influenced the CDP-choline pathway for phosphatidylcholine (PtdCho) biosynthesis. Relative to control CHO 7 cells, SRD 4 cells displayed increased PtdCho synthesis and degradation as indicated by a 4-6 fold increase in [(3)H]choline incorporation into PtdCho and 10-15-fold increase in intracellular [(3)H]glycerophosphocholine. [(3)H]Phosphocholine levels in SRD 4 cells were reduced by over 10-fold, suggesting enhanced activity of CTP:phosphocholine cytidylyltransferase alpha (CCTalpha). CHO-SCAP and CHO-SCAP D443N cells displayed modest increases in [(3)H]choline incorporation into PtdCho (2-fold) and only a 2-fold reduction in [(3)H]phosphocholine. Elevated PtdCho metabolism in SRD 4, compared with SCAP-overexpressing cells, was correlated with fatty acid synthesis. Inhibition of fatty acid synthesis by cerulenin resulted in almost complete normalization of PtdCho synthesis and choline metabolite profiles in SRD 4 cells, indicating that fatty acids or a fatty acid-derived metabolite was responsible for up-regulation of PtdCho synthesis. In contrast to apparent activation in vivo, CCTalpha protein, mRNA, and in vitro activity were reduced in SRD 4 cells and unchanged in SCAP transfected cells. Unlike control and SCAP transfected cells, CCTalpha in SRD 4 cells was localized by immunofluorescence to the nuclear envelope, suggesting that residual enzyme activity in these cells was in an active membrane-associated form. Translocation of CCTalpha to the nuclear envelope was reproduced by treatment of CHO 7 cells with exogenous oleate. We conclude that the SREBP/SCAP pathway regulates PtdCho synthesis via post transcriptional activation of nuclear CCTalpha by fatty acids or a fatty acid derived signal. PMID- 10799519 TI - Conversion of interleukin-13 into a high affinity agonist by a single amino acid substitution. AB - We created a novel mutated form of human interleukin-13 (IL-13) in which a positively charged arginine (R) at position 112 was substituted to a negatively charged aspartic acid (D). This mutant, termed IL-13R112D, was expressed in Escherichia coli and purified to near homogeneity. IL-13R112D was found to be a potent IL-13 agonist with 5-10-fold improved binding affinity to IL-13 receptors compared with wild-type IL-13 (wtIL-13). The conclusion of IL-13 agonist activity was drawn on the basis of approximately 10-fold improved activity over wtIL-13 in several assays: (a) inhibition of CD14 expression in primary monocytes; (b) proliferation of TF-1 and B9 cell lines; and (c) activation of STAT6 in Epstein Barr virus-immortalized B cells, primary monocytes, and THP-1 monocytic cell line. Furthermore, mutant IL-13R112D neutralized the cytotoxic activity of a chimeric fusion protein composed of wtIL-13 and a Pseudomonas exotoxin A (IL-13 PE38) approximately 10 times better than wtIL-13. Based on these results, it was concluded that IL-13R112D interacts with much stronger affinity than wtIL-13 on all cell types tested and that Arg-112 plays an important role in the interaction with its receptors (IL-13R). Thus, these results suggest that IL-13R112D may be a useful ligand for the study of IL-13 interaction with its receptors or, alternatively, in designing specific targeted agents for IL-13R-positive malignancies. PMID- 10799520 TI - Byr4 localizes to spindle-pole bodies in a cell cycle-regulated manner to control Cdc7 localization and septation in fission yeast. AB - Cytokinesis and septation in the fission yeast Schizosaccharomyces pombe are studied as a model for mammalian cell division. In fission yeast, septation is positively regulated by Spg1, a Ras family GTPase that localizes to spindle-pole bodies (SPBs) throughout the cell cycle. As cells enter mitosis, Spg1 accumulates in an active, GTP-bound form and binds the Cdc7 protein kinase to cause Cdc7 translocation to SPBs. Cdc7 disappears from one SPB in mid-anaphase and from the second SPB in late mitosis. Byr4 plus Cdc16 negatively regulate septation by forming a two-component GTPase-activating protein for Spg1. These results led us to hypothesize that Byr4 localization to SPBs regulated the nucleotide state of Spg1, due to its ability to form Spg1GAP activity with Cdc16 and thus the binding of Cdc7 to Spg1 at SPBs. To test this hypothesis, Byr4 localization was determined using indirect immunofluorescence. This analysis revealed that Byr4 was localized to SPBs that did not contain Cdc7. In byr4(-) mutants, Cdc7 localized to interphase SPBs and only symmetrically localized to mitotic SPBs. In contrast, Byr4 overexpression prevented Spg1 and Cdc7 localization to SPBs. These results suggest that Byr4 localization to SPBs maintains Spg1 in an inactive form, presumably by stimulating Spg1 GTPase activity with Cdc16, and that loss of Byr4 from mitotic SPBs increases the active fraction of Spg1 and thereby increases Spg1-Cdc7 binding. Byr4 localization to SPBs was decreased in spg1, cdc16, sid4, and cdc11 mutants as well as in several mutants that affect medial F actin structures, suggesting that multiple pathways regulate Byr4 localization to SPBs. PMID- 10799521 TI - Activation of peroxisome proliferator-activated receptor-gamma pathway inhibits osteoclast differentiation. AB - The nuclear receptor and transcription factor, peroxisome proliferator-activated receptor-gamma (PPAR-gamma), regulates the activity of other transcription factors in the adipogenic differentiation and inflammatory response pathways. We examined the possible function of the PPAR-gamma pathway in osteoclast (Ocl) formation from CD34(+) hematopoietic stem cells (CD34(+) HSCs), using a co culture system comprised of human mesenchymal stem cells (hMSCs) and CD34(+) HSCs, both derived from bone marrow. Ocl formation in this co-culture system is enhanced by the addition of exogenous osteoprotegerin ligand (OPGL), an essential Ocl differentiation factor, and macrophage-colony stimulating factor (M-CSF). The data indicate that soluble OPGL (sOPGL) and M-CSF stimulate Ocl formation in the co-cultures up to 4-fold compared with CD34(+) HSCs alone treated with sOPGL and M-CSF. CD34(+) HSCs, but not hMSCs, express PPAR-gamma, and 15-deoxy-Delta(12, 14)-prostaglandin-J2 (15d-PG-J2), a PPAR-gamma agonist, completely blocked the effects of sOPGL and M-CSF on Ocl formation and activity. The inhibitory effect of 15d-PG-J2 is specific to the Ocl lineage in both human and mouse models of osteoclastogenesis. Accordingly, parallel experiments demonstrate that sOPGL activates the NF-kappaB pathway within mouse Ocl progenitors, and this effect was abolished by 15d-PG-J2. These data establish a link between PPAR-gamma and OPGL signaling within Ocl progenitors, and support a role for PPAR-gamma pathway in the modulation of osteoclastogenesis. PMID- 10799522 TI - Molecular cloning and characterization of a novel ATP P2X receptor subtype from embryonic chick skeletal muscle. AB - We have cloned a new P2X ligand-gated ion channel receptor from embryonic chick skeletal muscle, which is tentatively named as chick P2X(8) (cP2X(8)) receptor. The cloned cDNA encodes a protein with 402 amino acids. Electrophysiological study of the recombinant cP2X(8) receptor expressed in Xenopus oocytes showed that 10 microm ATP induced a fast inward current followed by rapid and long lasting desensitization in medium containing 1.8 mm Ca(2+). In medium with 0. 3 mm Ca(2+) ATP induced a bi-phasic response as follows: a slower inward current succeeded the initial fast one. 2-Methylthio-ATP, alpha,beta-methylene-ATP, and adenosine 5'-O-(thio)triphosphate were potent agonists, whereas ADP was a very weak agonist. ATP-induced currents were blocked by 100 microm suramin and pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid. Northern blot analysis and reverse transcription-polymerase chain reaction showed that cP2X(8) RNA transcripts were mainly expressed in skeletal muscle, brain, and heart of Day 10 chick embryos. A moderate level of expression was also detected in gizzard and retina. Whole mount in situ hybridization showed that cP2X(8) RNA transcripts were expressed mainly in neurotube, notochord, and stomach in Day 3 embryos. In Day 4 and Day 6 embryos, the cP2X(8) RNA transcripts were highly expressed in the myotome and premuscle mass. The physiological role of this receptor in the establishment of the skeletal muscle innervation will be studied. PMID- 10799524 TI - Evidence for a novel Cdc42GAP domain at the carboxyl terminus of BNIP-2. AB - We recently identified BNIP-2, a previously cloned Bcl-2- and E1B-associated protein, as a putative substrate of the FGF receptor tyrosine kinase and showed that it possesses GTPase-activating activity toward Cdc42 despite the lack of homology to previously described catalytic domains of GTPase-activating proteins (GAPs). BNIP-2 contains many arginine residues at the carboxyl terminus, which includes the region of homology to the noncatalytic domain of Cdc42GAP, termed BNIP-2 and Cdc42GAP homology (BCH) domain. Using BNIP-2 glutathione S-transferase recombinants, it was found that its BCH bound Cdc42, and contributed the GAP activity. This domain was predicted to fold into alpha-helical bundles similar to the topology of the catalytic GAP domain of Cdc42GAP. Alignment of exposed arginine residues in this domain helped to identify Arg-235 and Arg-238 as good candidates for catalysis. Arg-238 matched well to the arginine "finger" required for enhanced GTP hydrolysis in homodimerized Cdc42. Site-directed mutagenesis confirmed that an R235K or R238K mutation severely impaired the BNIP-2 GAP activity without affecting its binding to Cdc42. From deletion studies, a region adjacent to the arginine patch ((288)EYV(290) on BNIP-2) and the Switch I and Rho family-specific "Insert" region on Cdc42 are involved in the binding. The results indicate that the BCH domain of BNIP-2 represents a novel GAP domain that employs an arginine patch motif similar to that of the Cdc42-homodimer. PMID- 10799523 TI - Nitrogen catabolite repression of DAL80 expression depends on the relative levels of Gat1p and Ure2p production in Saccharomyces cerevisiae. AB - GATA family activators (Gln3p and Gat1p) and repressors (Dal80p and Deh1p) regulate nitrogen catabolite repression (NCR)-sensitive transcription in Saccharomyces cerevisiae presumably via their competitive binding to the GATA sequences upstream of NCR-sensitive genes. Ure2p, which is not a GATA family member, inhibits Gln3p/Gat1p from functioning in the presence of good nitrogen sources. We show that NCR-sensitive DAL80 transcription can be influenced by the relative levels of GAT1 and URE2 expression. NCR, normally observed with ammonia or glutamine, is severely diminished when Gat1p is overproduced, and this inhibition is overcome by simultaneously increasing URE2 expression. Further, overproduction of Ure2p nearly eliminates NCR-sensitive transcription under derepressive growth conditions, i.e. with proline as the sole nitrogen source. Enhanced green fluorescent protein-Gat1p is nuclear when Gat1p-dependent transcription is high and cytoplasmic when it is inhibited by overproduction of Ure2p. PMID- 10799525 TI - Bivalent sequential binding model of a Bacillus thuringiensis toxin to gypsy moth aminopeptidase N receptor. AB - Specificity for target insects of Bacillus thuringiensis insecticidal Cry toxins is largely determined by toxin affinity for insect midgut receptors. The mode of binding for one such toxin-receptor complex was investigated by extensive toxin mutagenesis, followed by real-time receptor binding analysis using an optical biosensor (BIAcore). Wild-type Cry1Ac, a three-domain, lepidopteran-specific toxin, bound purified gypsy moth (Lymantria dispar) aminopeptidase N (APN) biphasically. Site 1 displayed fast association and dissociation kinetics, while site 2 possessed slower kinetics, yet tighter affinity. We empirically determined that two Cry1Ac surface regions are involved in in vivo toxicity and APN binding. Mutations within domain III affected binding rates to APN site 1, whereas mutations in domain II affected binding rates to APN site 2. Furthermore, domain III contact is completely inhibited in the presence of N-acetylgalactosamine, indicating loss of domain III binding eliminates all APN binding. Based upon these observations, the following model is proposed. A cavity in lectin-like domain III initiates docking through recognition of an N-acetylgalactosamine moiety on L. dispar APN. Following primary docking, a higher affinity domain II binding mechanism occurs, which is critical for insecticidal activity. PMID- 10799526 TI - A map of protein-rRNA distribution in the 70 S Escherichia coli ribosome. AB - Neutron scattering exploits the enormous scattering difference between protons and deuterons. A set of 42 x-ray and neutron solution scattering curves from hybrid Escherichia coli ribosomes was obtained, where the proteins and rRNA moieties in the subunits were either protonated or deuterated in all possible combinations. This extensive data set is analyzed using a novel method. The volume defined by the cryoelectron microscopic model of Frank and co-workers (Frank, J., Zhu, J., Penczek, P., Li, Y. H., Srivastava, S., Verschoor, A., Radermacher, M., Grassucci, R., Lata, R. K., and Agrawal, R. K. (1995) Nature 376, 441-444) is divided into 7890 densely packed spheres of radius 0.5 nm. Simulated annealing is employed to assign each sphere to solvent, protein, or rRNA moieties to simultaneously fit all scattering curves. Twelve independent reconstructions starting from random approximations yielded reproducible results. The resulting model at a resolution of 3 nm represents the volumes occupied by rRNA and protein moieties at 95% probability threshold and displays 15 and 20 protein subvolumes in the 30 S and 50 S, respectively, connected by rRNA. 17 proteins with known atomic structure can be tentatively positioned into the protein subvolumes within the ribosome in agreement with the results from other methods. The protein-rRNA map enlarges the basis for the models of the rRNA folding and can further help to localize proteins in high-resolution crystallographic density maps. PMID- 10799527 TI - A new gene involved in the transport-dependent metabolism of phosphatidylserine, PSTB2/PDR17, shares sequence similarity with the gene encoding the phosphatidylinositol/phosphatidylcholine transfer protein, SEC14. AB - A new yeast strain, designated pstB2, that is defective in the conversion of nascent phosphatidylserine (PtdSer) to phosphatidylethanolamine (PtdEtn) by PtdSer decarboxylase 2, has been isolated. The pstB2 strain requires ethanolamine for growth. Incubation of cells with [(3)H]serine followed by analysis of the aminoglycerophospholipids demonstrates a 50% increase in the labeling of PtdSer and a 72% decrease in PtdEtn formation in the mutant relative to the parental strain. The PSTB2 gene was isolated by complementation, and it restores ethanolamine prototrophy and corrects the defective lipid metabolism of the pstB2 strain. The PSTB2 gene is allelic to the pleiotropic drug resistance gene, PDR17, and is homologous to SEC14, which encodes a phosphatidylinositol/phosphatidylcholine transfer protein. The protein, PstB2p, displays phosphatidylinositol but not PtdSer transfer activity, and its overexpression causes suppression of sec14 mutants. However, overexpression of the SEC14 gene fails to suppress the conditional lethality of pstB2 strains. The transport-dependent metabolism of PtdSer to PtdEtn occurs in permeabilized wild type yeast but is dramatically reduced in permeabilized pstB2 strains. Fractionation of permeabilized cells demonstrates that the pstB2 strain accumulates nascent PtdSer in the Golgi apparatus and a novel light membrane fraction, consistent with a defect in lipid transport processes that control substrate access to PtdSer decarboxylase 2. PMID- 10799528 TI - A 56-kDa selenium-binding protein participates in intra-Golgi protein transport. AB - Transport of proteins between intracellular membrane compartments is a highly regulated process that depends on several cytosolic factors. By using the well characterized intra-Golgi cell-free transport assay, we purified from bovine brain cytosol a 56-kDa protein that shows a significant transport activity. Partial sequencing of four tryptic peptides obtained from the 56-kDa protein revealed its identity to a cytosolic protein previously characterized as a selenium-binding protein, SBP56. Recombinant SBP56 expressed in Escherichia coli exhibited transport activity when added to the cell-free intra-Golgi transport. Affinity purified anti-SBP56 polyclonal antibodies specifically inhibited intra Golgi transport in vitro. Although SBP56 is predominantly localized in the cytosol, a significant amount is associated with membranes. Subcellular fractionation showed that this protein is peripherally associated with the Golgi membrane. The experiments presented in this study indicate that SBP56 participates in late stages of intra-Golgi protein transport. PMID- 10799529 TI - The Akt-glycogen synthase kinase 3beta pathway regulates transcription of atrial natriuretic factor induced by beta-adrenergic receptor stimulation in cardiac myocytes. AB - We examined the mechanism of atrial natriuretic factor (ANF) transcription by isoproterenol (ISO), an agonist for the beta-adrenergic receptor (betaAR), in cardiac myocytes. ISO only modestly activated members of the mitogen-activated protein kinase family. ISO-induced ANF transcription was not affected by inhibition of mitogen-activated protein kinases, whereas it was significantly inhibited by KN93, an inhibitor of Ca(2+)/calmodulin-dependent kinase (CaM kinase II). Production of 3'-phosphorylated phosphatidylinositides (3 phosphoinositides) was also required for ISO-induced ANF transcription. ISO caused phosphorylation (Ser-473) and activation of Akt through CaM kinase II- and 3 phosphoinositides dependent mechanisms. Constitutively active Akt increased myocyte surface area, total protein content, and ANF expression, whereas dominant negative Akt blocked ISO-stimulated ANF transcription. ISO caused Ser-9 phosphorylation and decreased activities of GSK3beta. Overexpression of GSK3beta inhibited ANF transcription, which was reversed by ISO. ISO failed to reverse the inhibitory effect of GSK3beta(S9A), an Akt-insensitive mutant. Kinase-inactive GSK3beta increased ANF transcription. Cyclosporin A partially inhibited ISO-stimulated ANF transcription, indicating that calcineurin only partially mediates ANF transcription. These results suggest that both CaM kinase II and 3 phosphoinositides mediate betaAR-induced Akt activation and ANF transcription in cardiac myocytes. Furthermore, betaAR-stimulated ANF transcription is predominantly mediated by activation of Akt and subsequent phosphorylation/inhibition of GSK3beta. PMID- 10799530 TI - Absence of the gamma subunit of the skeletal muscle dihydropyridine receptor increases L-type Ca2+ currents and alters channel inactivation properties. AB - In skeletal muscle the oligomeric alpha(1S), alpha(2)/delta-1 or alpha(2)/delta 2, beta1, and gamma1 L-type Ca(2+) channel or dihydropyridine receptor functions as a voltage sensor for excitation contraction coupling and is responsible for the L-type Ca(2+) current. The gamma1 subunit, which is tightly associated with this Ca(2+) channel, is a membrane-spanning protein exclusively expressed in skeletal muscle. Previously, heterologous expression studies revealed that gamma1 might modulate Ca(2+) currents expressed by the pore subunit found in heart, alpha(1C), shifting steady state inactivation, and increasing current amplitude. To determine the role of gamma1 assembled with the skeletal subunit composition in vivo, we used gene targeting to establish a mouse model, in which gamma1 expression is eliminated. Comparing litter-matched mice with control mice, we found that, in contrast to heterologous expression studies, the loss of gamma1 significantly increased the amplitude of peak dihydropyridine-sensitive I(Ca) in isolated myotubes. Whereas the activation kinetics of the current remained unchanged, inactivation of the current was slowed in gamma1-deficient myotubes and, correspondingly, steady state inactivation of I(Ca) was shifted to more positive membrane potentials. These results indicate that gamma1 decreases the amount of Ca(2+) entry during stimulation of skeletal muscle. PMID- 10799531 TI - An ordered array of cold shock domain repressor elements across tumor necrosis factor-responsive elements of the granulocyte-macrophage colony-stimulating factor promoter. AB - The tumor necrosis factor-alpha-responsive region of the human granulocyte macrophage colony-stimulating factor (GM-CSF) promoter (-114 to -31) encompasses binding sites for NF-kappaB, CBF, AP-1, ETS, and NFAT families of transcription factors. We show both here and previously that mutation of any one of these binding sites greatly reduces tumor necrosis factor-alpha induction of the GM-CSF promoter. Interspersed between these elements are sequences that when mutated lead to an increase in GM-CSF promoter activity. We have previously shown that two of these repressor elements bind proteins known as cold shock domain (CSD) factors and that overexpression of CSD proteins leads to repression of GM-CSF promoter activity in fibroblasts. CSD proteins are single strand DNA- and RNA binding proteins that contact 5'-CCTG-3' sequences in the GM-CSF repressor elements. We show here that two newly identified repressor sequences in the proximal promoter can also bind CSD proteins. We have characterized the CSD containing protein complexes that bind to the GM-CSF promoter and identified a novel protein related to mitochondrial single strand binding protein that forms part of one of these complexes. The four CSD-binding sites on the promoter occur in pairs on opposite strands of the DNA and appear to form an ordered array of binding elements. A similar ordered array of CSD sites are present in the promoters of the granulocyte colony-stimulating factor and interleukin-3 genes, implying a common mechanism for negative regulation of these myeloid growth factors. PMID- 10799532 TI - The phosphorylation state and expression of soybean BiP isoforms are differentially regulated following abiotic stresses. AB - The mammalian BiP is regulated by phosphorylation, and it is generally accepted that its unmodified form constitutes the biologically active species. In fact, the glycosylation inhibitor tunicamycin induces dephosphorylation of mammalian BiP. The stress-induced phosphorylation state of plant BiP has not been examined. Here, we demonstrated that soybean BiP exists in interconvertible phosphorylated and nonphosphorylated forms, and the equilibrium can be shift to either direction in response to different stimuli. In contrast to tunicamycin treatment, water stress condition stimulated phosphorylation of BiP species in soybean cultured cells and stressed leaves. Despite their phosphorylation state, we demonstrated that BiP isoforms from water-stressed leaves exhibit protein binding activity, suggesting that plant BiP functional regulation may differ from other eukaryotic BiPs. We also compared the induction of the soybean BiP gene family, which consists of at least four members designated soyBiPA, soyBiPB, soyBiPC, and soyBiPD, by tunicamycin and osmotic stress. Although all soybean BiP genes were induced by tunicamycin, just the soyBiPA RNA was up-regulated by osmotic stress. In addition, these stresses promoted BiP induction with different kinetics and acted synergistically to increase BiP accumulation. These results suggest that the soybean BiP gene family is differentially regulated by abiotic stresses through distinct signaling pathways. PMID- 10799533 TI - Acute regulation of fatty acid uptake involves the cellular redistribution of fatty acid translocase. AB - We used muscle contraction, which increases fatty acid oxidation, as a model to determine whether fatty acid transport is acutely regulated by fatty acid translocase (FAT/CD36). Palmitate uptake by giant vesicles, obtained from skeletal muscle, was increased by muscle contraction. Kinetic studies indicated that muscle contraction increased V(max), but K(m) remained unaltered. Sulfo-N succinimidyl oleate, a specific inhibitor of FAT/CD36, fully blocked the contraction-induced increase in palmitate uptake. In giant vesicles from contracting muscles, plasma membrane FAT/CD36 was also increased in parallel with the increase in long chain fatty acid uptake. Further studies showed that like GLUT-4, FAT/CD36 is located in both the plasma membrane and intracellularly (endosomally). With muscle contraction, FAT/CD36 at the surface of the muscle was increased, while concomitantly, FAT/CD36 in the intracellular pool was reduced. Similar responses were observed for GLUT-4. We conclude that fatty acid uptake is subject to short term regulation by muscle contraction and involves the translocation of FAT/CD36 from intracellular stores to the sarcolemma, analogous to the regulation of glucose uptake by GLUT-4. PMID- 10799534 TI - Heterogeneous nuclear ribonucleoprotein A1 and UP1 protect mammalian telomeric repeats and modulate telomere replication in vitro. AB - The heterogeneous nuclear ribonucleoprotein A1 protein and a shortened derivative (UP1) promote telomere elongation in mammalian cells. To gain insights into the function of A1/UP1 in telomere biogenesis, we have investigated the binding properties of recombinant A1/UP1 and derivatives to single-stranded DNA oligonucleotides. Our results indicate that UP1 prefers to bind to DNA carrying single-stranded telomeric extensions at the 3' terminus. The RNA recognition motif 1 is sufficient for strong and specific binding to oligomers carrying vertebrate telomeric repeats. We find that the binding of A1/UP1 protects telomeric sequences against degradation by endo- and exonucleases. Moreover, A1/UP1 binding prevents extension by telomerase and terminal deoxynucleotidyltransferase and inhibits rNTP-dependent DNA synthesis in vitro. These observations are consistent with the hypothesis that A1/UP1 is a telomere end-binding protein that plays a role in the maintenance of long 3' overhangs. PMID- 10799535 TI - Identification of a major heparin and cell binding site in the LG4 module of the laminin alpha 5 chain. AB - The G domain of the laminin alpha chains consists of five homologous G modules (LG1-5) and has been implicated in various biological functions. In this study, we identified an active site for cell and heparin binding within the laminin alpha5 G domain using recombinant proteins and synthetic peptides. Recombinant LG4, LG5, and LG4-5 modules were generated using a mammalian expression system. The LG4 and LG4-5 modules were highly active for cell binding, whereas the LG5 module alone showed only weak binding. Heparin inhibited cell binding to the LG4 5 module, whereas no inhibition was observed with EDTA or antibodies against the integrin beta(1) subunit. These results suggest that the LG4-5 module interacts with a cell surface receptor containing heparan sulfate but not with integrins. Solid-phase assays and surface plasmon resonance measurements demonstrated strong binding of the LG4 and LG4-5 modules to heparin with K(D) values in the nanomolar range, whereas a 16-fold lower value was determined for the LG5 module. Treatment with glycosidases demonstrated that N-linked carbohydrates on the LG5 module are complex-type oligosaccharides. The LG4-5 module, devoid of N-linked carbohydrates, exhibited similar binding kinetics toward heparin. Furthermore, cell binding was unaffected by removal of N-linked glycosylation. To localize active sites on the LG4 module, various synthetic peptides were used to compete with binding of the tandem module to heparin and cells. Peptide F4 (AGQWHRVSVRWG) inhibited binding, whereas a scrambled peptide of F4 failed to compete binding. Alanine replacements demonstrated that one arginine residue within F4 was important for cell and heparin binding. Our results suggest a critical role of the LG4 module for heparan sulfate-containing receptor binding within the laminin alpha5 chain. PMID- 10799536 TI - Involvement of thyrotroph embryonic factor in calcium-mediated regulation of gene expression. AB - In the present study, we used an expression cloning strategy to identify transcription factors that bind specifically to a limited region of the inducible cAMP early repressor (ICER) promoter and regulate transcription. Murine thyrotroph embryonic factor (mTEF) was isolated and was shown to bind to a site located at nucleotides -117 to -108 from the transcriptional start site. Transient expression of reporter constructs containing either a consensus TEFRE or the icerTEF binding site demonstrated that TEF-dependent transcription correlated with relative binding affinities, i.e. the consensus TEFRE bound TEF more tightly and was more responsive to TEF than the icerTEFRE. Because the icerTEFRE overlapped a cAMP response element, the responsiveness of these sequences to either cAMP or Ca(2+) was tested. Although TEF expression had no effect on the cAMP-regulated transcriptional response of the ICER promoter, TEF did confer calcium responsiveness to these sequences. Calcium also modestly increased the TEF-mediated transcription from a consensus TEFRE. Additional studies using Ca(2+)-activated kinases indicate that Ca(2+)/TEF/TEFRE-regulated transcription may be mediated through Ca(2+)/calmodulin-dependent kinase (CaMK) IV. Moreover, studies with the icerTEFRE in a CaMK IV-deficient cell line demonstrated that these cells were transcriptionally unresponsive to thapsigargin; however, responsiveness was restored by co-expression of the active CaMK IV. These studies are the first to demonstrate that TEF is a calcium responsive transcription factor, and they suggest that there are two classes of TEF-regulated genes. One class, represented by a consensus TEFRE, is regulated by TEF in the resting cell; the second class, represented by icerTEFRE, is regulated by TEF in the calcium-activated cell. PMID- 10799537 TI - The collagen-like peptide (GER)15GPCCG forms pH-dependent covalently linked triple helical trimers. AB - A collagen-like peptide with the sequence (GER)(15) GPCCG was synthesized to study the formation of a triple helix in the absence of proline residues. This peptide can form a triple helix at acidic and basic pH, but is insoluble around neutral pH. The formation of a triple helix can be used to covalently oxidize the cysteine residues into a disulfide knot. Three disulfide bonds are formed between the three chains as has been found at the carboxyl-terminal end of the type III collagen triple helix. This is a new method to covalently link collagen-like peptides with a stereochemistry that occurs in nature. The peptide undergoes a reversible, cooperative triple helix coil transition with a transition midpoint (T(m)) of 17 to 20 degrees C at acidic pH and 32 to 37 degrees C at basic pH. At acidic pH there was little influence of the T(m) on the salt concentration of the buffer. At basic pH increasing the salt concentration reduced the T(m) to values comparable to the stability at acidic pH. These experiments show that the tripeptide unit GER which occurs frequently in collagen sequences can form a triple helical structure in the absence of more typical collagen-like tripeptide units and that charge-charge interactions play a role in the stabilization of the triple helix of this peptide. PMID- 10799538 TI - Characterization of a P-type Na+-ATPase of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum. AB - A facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum, possesses a P type Na(+)-stimulated ATPase in the membrane (Koyama, N. (1999) Curr. Microbiol. 39, 27-30). In this study, we attempted to purify and characterize the enzyme. The ATPase appears to consist of a single polypeptide with an apparent molecular mass of 100 kDa. The enzyme exhibited an optimum pH for activity at around 9. The enzyme was strongly inhibited by vanadate (50% inhibition observed at 3 microm) and forms an acylphosphate intermediate, suggesting a P-type ATPase. The enzyme, when reconstituted into soybean phospholipid vesicles, exhibited ATP-dependent (22)Na(+) uptake, which was completely inhibited by gramicidin. The reconstituted vesicles exhibited a generation of membrane potential (positive, inside). The enzyme is likely to be involved in an electrogenic transport of Na(+). PMID- 10799539 TI - Overproduction in Escherichia coli and characterization of yeast replication factor C lacking the ligase homology domain. AB - Eukaryotic replication factor C (RF-C) is a heteropentameric complex that is required to load the replication clamp proliferating cell nuclear antigen onto primed DNA. Saccharomyces cerevisiae RF-C is encoded by the genes RFC1-RFC5. The RFC1 gene was cloned under control of the strong inducible bacteriophage T7 promoter, yet induction did not yield detectable Rfc1p. However, a truncated form of RFC1 deleted for the coding region for amino acids 3-273, rfc1-DeltaN, did allow overproduction. The other four RFC genes were cloned into the latter plasmid to yield a single plasmid that overproduced RF-C to moderate levels. Overproduction of the complex was further enhanced when the Escherichia coli argU gene encoding the rare arginine tRNA was also overproduced. The enzyme thus produced in E. coli was purified to homogeneity through three column steps, including a proliferating cell nuclear antigen affinity column. This enzyme, as well as the enzyme purified from yeast, is prone to aggregation and inactivation, and therefore, light scattering was used to determine conditions stabilizing the enzyme and preventing aggregation. Broad-range carrier ampholytes at about 0.05% were found to be most effective. In some assays, the Rfc1-DeltaN containing RF-C from E. coli showed an increased activity compared with the full-length enzyme from yeast, likely because the latter enzyme exhibits significant nonspecific binding to single-stranded DNA. Replacement of RFC1 by rfc1-DeltaN in yeast shows essentially no phenotype with regard to DNA replication, damage susceptibility, telomere length maintenance, and intrachromosomal recombination. PMID- 10799540 TI - Mammalian Sec61 is associated with Sec62 and Sec63. AB - In yeast, efficient protein transport across the endoplasmic reticulum (ER) membrane may occur co-translationally or post-translationally. The latter process is mediated by a membrane protein complex that consists of the Sec61p complex and the Sec62p-Sec63p subcomplex. In contrast, in mammalian cells protein translocation is almost exclusively co-translational. This transport depends on the Sec61 complex, which is homologous to the yeast Sec61p complex and has been identified in mammals as a ribosome-bound pore-forming membrane protein complex. We report here the existence of ribosome-free mammalian Sec61 complexes that associate with two ubiquitous proteins of the ER membrane. According to primary sequence analysis both proteins display homology to the yeast proteins Sec62p and Sec63p and are therefore named Sec62 and Sec63, respectively. The probable function of the mammalian Sec61-Sec62-Sec63 complex is discussed with respect to its abundance in ER membranes, which, in contrast to yeast ER membranes, apparently lack efficient post-translational translocation activity. PMID- 10799541 TI - C1 inhibitor cross-linking by tissue transglutaminase. AB - C1 inhibitor, a plasma proteinase inhibitor of the serpin superfamily involved in the regulation of complement classical pathway and intrinsic blood coagulation, has been shown to bind to several components of the extracellular matrix. These reactions may be responsible for C1 inhibitor localization in the perivascular space. In the study reported here, we have examined whether C1 inhibitor could function as a substrate for plasma (factor XIIIa) or tissue transglutaminase. We made the following observations: 1) SDS-polyacrylamide gel electrophoresis and autoradiography showed that C1 inhibitor exposed to tissue transglutaminase (but not to factor XIIIa) incorporated the radioactive amine donor substrate [(3)H]putrescine in a calcium-dependent manner; 2) the maximum stoichiometry for the uptake of [(3)H]putrescine by C1 inhibitor was 1:1; 3) proteolytic cleavage and peptide sequencing of reduced and carboxymethylated [(3)H]putrescine-C1 inhibitor identified Gln(453) (P'9) as the single amine acceptor residue; 4) studies with (125)I-labeled C1 inhibitor showed that tissue transglutaminase was also able to cross-link C1 inhibitor to immobilized fibrin; and 5) C1 inhibitor cross-linked by tissue transglutaminase to immobilized fibrin had inhibitory activity against its target enzymes. Thus, tissue transglutaminase-mediated cross linking of C1 inhibitor to fibrin or other extracellular matrix components may serve as a mechanism for covalent serpin binding and influence local regulation of the proteolytic pathways inhibited by C1 inhibitor. PMID- 10799542 TI - Activation of downstream signals by the long form of the leptin receptor. AB - The adipocyte-derived hormone leptin signals the status of body energy stores by activating the long form of the leptin receptor (LRb). Activation of LRb results in the activation of the associated Jak2 tyrosine kinase and the transmission of downstream phosphotyrosine-dependent signals. We have investigated the signaling function of mutant LRb intracellular domains under the control of the extracellular erythropoietin (Epo) receptor. By using this system, we confirm that two tyrosine residues in the intracellular domain of murine LRb become phosphorylated to mediate LRb signaling; Tyr(985) controls the tyrosine phosphorylation of SHP-2, and Tyr(1138) controls STAT3 activation. We furthermore investigated the mechanisms by which LRb controls downstream ERK activation and c fos and SOCS3 message accumulation. Tyr(985)-mediated recruitment of SHP-2 does not alter tyrosine phosphorylation of Jak2 or STAT3 but results in GRB-2 binding to tyrosine-phosphorylated SHP-2 and is required for the majority of ERK activation during LRb signaling. Tyr(985) and ERK activation similarly mediate c fos mRNA accumulation. In contrast, SOCS3 mRNA accumulation requires Tyr(1138) mediated STAT3 activation. Thus, the two LRb tyrosine residues that are phosphorylated during receptor activation mediate distinct signaling pathways as follows: SHP-2 binding to Tyr(985) positively regulates the ERK --> c-fos pathway, and STAT3 binding to Tyr(1138) mediates the inhibitory SOCS3 pathway. PMID- 10799543 TI - Gelsolin binding and cellular presentation of lysophosphatidic acid. AB - Lysophosphatidic acid (LPA) in biological fluids binds to serum albumin and other proteins that enhance its effects on cellular functions. The actin-severing protein gelsolin binds LPA with an affinity (K(d) = 6 nm) similar to that of the G protein-coupled LPA receptors encoded by endothelial differentiation genes 2, 4, and 7 (Edg-2, -4, and -7 receptors) and greater than that of serum albumin (K(d) = 360 nm). At concentrations of 10% or less of that in plasma, which are observed in fluids of injured tissues, purified and recombinant gelsolin augment LPA stimulation of nuclear signals and protein synthesis in rat cardiac myocytes (RCMs) that express Edg-2 and -4 receptors. At concentrations of 20% or more of that in plasma, gelsolin suppresses LPA stimulation of RCMs. The lack of effect of gelsolin on RCM responses to monoclonal anti-Edg-4 receptor antibody plus a phorbol ester without LPA attests to its specificity for LPA delivery and the absence of post-receptor effects. Inhibition of gelsolin binding and cellular delivery of LPA by l-alpha-phosphatidylinositol-4,5-bisphosphate (PIP2) and peptides constituting the two PIP2 binding domains of gelsolin suggests competition between LPA and PIP2 for the same sites. Thus, delivery of LPA to RCMs is affinity-coupled to Edg receptors by gelsolin in a PIP2-regulated process. PMID- 10799544 TI - Streptokinase binds to human plasmin with high affinity, perturbs the plasmin active site, and induces expression of a substrate recognition exosite for plasminogen. AB - Binding of streptokinase (SK) to plasminogen (Pg) conformationally activates the zymogen and converts both Pg and plasmin (Pm) into specific Pg activators. The interaction of SK with Pm and its relationship to the mechanism of Pg activation were evaluated in equilibrium binding studies with active site-labeled fluorescent Pm derivatives and in kinetic studies of SK-induced changes in the catalytic specificity of Pm. SK bound to fluorescein-labeled and native Pm with dissociation constants of 11 +/- 2 pm and 12 +/- 4 pm, which represented a 1,000 10,000-fold higher affinity than determined for Pg. Stoichiometric binding of SK to native Pm was followed by generation of a two-fragment form of SK cleaved at Lys(59) (SK'), which exhibited an indistinguishable affinity for labeled Pm, while a truncated, SK(55-414) species had a 120-360-fold reduced affinity. Binding of SK to native Pm was accompanied by a >50-fold enhancement in specificity for activation of Pg, which was paralleled by a surprising 2.6-10 fold loss of specificity of Pm for 8 of 11 tripeptide-pNA substrates. Further studies with Pm labeled at the active site with 2-anilinonaphthalene-6-sulfonic acid demonstrated directly that binding of SK to Pm resulted in expression of a new substrate binding exosite for Pg on the SK.Pm complex. It is concluded that SK activates Pg in part by preferential binding to the active zymogen conformation. High affinity binding of SK to Pm enhances Pg substrate specificity principally through emergence of a substrate recognition exosite. PMID- 10799545 TI - Evidence that Llck-mediated phosphorylation of p56dok and p62dok may play a role in CD2 signaling. AB - The Lck tyrosine kinase is involved in signaling by T cell surface receptors such as TCR/CD3, CD2, and CD28. As other downstream protein-tyrosine kinases are activated upon stimulation of these receptors, it is difficult to assign which tyrosine-phosphorylated proteins represent bona fide Lck substrates and which are phosphorylated by other tyrosine kinases. We have developed a system in which Lck can be activated independently of TCR/CD3. We have shown that activation of an epidermal growth factor receptor/Lck chimera leads to the specific phosphorylation of Ras GTPase-activating protein (RasGAP) and two RasGAP associated proteins, p56(dok) and p62(dok). Activation of the chimeric protein correlates with an increase in cellular Ca(2+) in the absence of ZAP-70 and phospholipase Cgamma1 phosphorylation. Furthermore, we have found that p62(dok) co-immunoprecipitates with the activated epidermal growth factor receptor/LckF505 and that phosphorylated Dok proteins bind to the Src homology 2 domain of Lck in vitro. In addition, we have shown that activation via the CD2 but not the TCR/CD3 receptor leads to the phosphorylation of p56(dok) and p62(dok). Using JCaM1.6 cells, we have demonstrated that Lck is required for CD2-mediated phosphorylation of Dok proteins. We propose that phosphorylation and Src homology 2-mediated association of p56(dok) and p62(dok) with Lck play a selective function in accessory receptor signal transduction mechanisms. PMID- 10799546 TI - Stimulation-induced down-regulation of tumor necrosis factor-alpha converting enzyme. AB - The extracellular domains of many proteins, including growth factors, cytokines, receptors, and adhesion molecules, are proteolytically released from cells, a process termed "shedding." Tumor necrosis factor-alpha converting enzyme (TACE/ADAM-17) is a metalloprotease-disintegrin that sheds tumor necrosis factor alpha and other proteins. To study the regulation of TACE-mediated shedding, we examined the effects of stimulation of cells on TACE localization and expression. Immunofluorescence microscopy revealed a punctate distribution of TACE on the surface of untreated cells, and stimulation of monocytic cells with lipopolysaccharide did not affect TACE staining. Phorbol 12-myristate 13-acetate (PMA), a potent inducer of shedding, decreased cell-surface staining for TACE. Surface biotinylation experiments confirmed and extended this observation; PMA decreased the half-life of surface-biotinylated TACE without increasing the turnover of total cell-surface proteins. Soluble fragments of TACE were not detected in the medium of cells that had down-regulated TACE, and TACE was not down-regulated when endocytosis was inhibited. Antibody uptake experiments suggested that cell-surface TACE was internalized in response to PMA. Surprisingly, a metalloprotease inhibitor prevented the PMA-induced turnover of TACE. Thus, PMA activates shedding and causes the down-regulation of a major "sheddase," suggesting that induced shedding may be regulated by a mechanism that decreases the amount of active TACE on the cell surface. PMID- 10799547 TI - Functional analysis of the domain structure of tumor necrosis factor-alpha converting enzyme. AB - Many membrane-bound proteins, including cytokines, receptors, and growth factors, are proteolytically cleaved to release a soluble form of their extracellular domain. The tumor necrosis factor (TNF)-alpha converting enzyme (TACE/ADAM-17) is a transmembrane metalloproteinase responsible for the proteolytic release or "shedding" of several cell-surface proteins, including TNF and p75 TNFR. We established a TACE-reconstitution system using TACE-deficient cells co transfected with TACE and substrate cDNAs to study TACE function and regulation. Using the TACE-reconstitution system, we identified two additional substrates of TACE, interleukin (IL)-1R-II and p55 TNFR. Using truncations and chimeric constructs of TACE and another ADAM family member, ADAM-10, we studied the function of the different domains of TACE in three shedding activities. We found that TACE must be expressed with its membrane-anchoring domain for phorbol ester stimulated shedding of TNF, p75 TNFR, and IL-1R-II, but that the cytoplasmic domain is not required for the shedding of these substrates. The catalytic domain of ADAM-10 could not be functionally substituted for that of TACE. IL-1R-II shedding required the cysteine-rich domain of TACE as well as the catalytic domain, whereas TNF and p75 TNFR shedding required only the tethered TACE catalytic domain. PMID- 10799548 TI - Hck enhances the adherence of lipopolysaccharide-stimulated macrophages via Cbl and phosphatidylinositol 3-kinase. AB - Src family tyrosine kinases have previously been proposed to mediate some of the biological effects of lipopolysaccharide on macrophages. Accordingly, we have sought to identify substrates of Src family kinases in lipopolysaccharide stimulated macrophages. Stimulation of Bac1.2F5 macrophage cells with lipopolysaccharide was found to induce gradual and persistent tyrosine phosphorylation of Cbl in an Src family kinase-dependent manner. Immunoprecipitation experiments revealed that Cbl associates with Hck in Bac1.2F5 cells, while expression of an activated form of Hck in Bac1.2F5 cells induces tyrosine phosphorylation of Cbl in the absence of lipopolysaccharide stimulation. The Src homology 3 domain of Hck can directly bind Cbl, and this interaction is important for phosphorylation of Cbl. Association of the p85 subunit of phosphatidylinositol (PI) 3-kinase with Cbl is enhanced following lipopolysaccharide stimulation of Bac1.2F5 cells, and transient expression experiments indicate that phosphorylation of Cbl by Hck can facilitate the association of p85 with Cbl. Lipopolysaccharide treatment also stimulates the partial translocation of Hck to the cytoskeleton of Bac1.2F5 cells. Notably, lipopolysaccharide enhances the adherence of Bac1.2F5 cells, an effect that is dependent on the activity of Src family kinases and PI 3-kinase. Thus, we postulate that Hck enhances the adherence of lipopolysaccharide-stimulated macrophages, at least in part, via Cbl and PI 3-kinase. PMID- 10799549 TI - Epidermal growth factor receptor-dependent Akt activation by oxidative stress enhances cell survival. AB - The serine/threonine kinase Akt (also known as protein kinase B) is activated in response to various stimuli by a mechanism involving phosphoinositide 3-kinase (PI3-K). Akt provides a survival signal that protects cells from apoptosis induced by growth factor withdrawal, but its function in other forms of stress is less clear. Here we investigated the role of PI3-K/Akt during the cellular response to oxidant injury. H(2)O(2) treatment elevated Akt activity in multiple cell types in a time- (5-30 min) and dose (400 microM-2 mm)-dependent manner. Expression of a dominant negative mutant of p85 (regulatory component of PI3-K) and treatment with inhibitors of PI3-K (wortmannin and LY294002) prevented H(2)O(2)-induced Akt activation. Akt activation by H(2)O(2) also depended on epidermal growth factor receptor (EGFR) signaling; H(2)O(2) treatment led to EGFR phosphorylation, and inhibition of EGFR activation prevented Akt activation by H(2)O(2). As H(2)O(2) causes apoptosis of HeLa cells, we investigated whether alterations of PI3-K/Akt signaling would affect this response. Wortmannin and LY294002 treatment significantly enhanced H(2)O(2)-induced apoptosis, whereas expression of exogenous myristoylated Akt (an activated form) inhibited cell death. Constitutive expression of v-Akt likewise enhanced survival of H(2)O(2) treated NIH3T3 cells. These results suggest that H(2)O(2) activates Akt via an EGFR/PI3-K-dependent pathway and that elevated Akt activity confers protection against oxidative stress-induced apoptosis. PMID- 10799550 TI - Factor VIIa and thrombin induce the expression of Cyr61 and connective tissue growth factor, extracellular matrix signaling proteins that could act as possible downstream mediators in factor VIIa x tissue factor-induced signal transduction. AB - Extracellular interactions of plasma clotting factor VIIa (FVIIa) with tissue factor (TF) on cell surfaces trigger the intracellular signaling events. At present, it is unclear how these signals influence phenotype. To elucidate this, we have used cDNA microarray technology to examine changes in transcriptional program in human fibroblasts in response to exposure to FVIIa. cDNA microarrays revealed that FVIIa binding to TF up-regulated the expression of Cyr61 and CTGF (connective tissue growth factor), the genes that encode extracellular matrix signaling proteins Cyr61 and CTGF, respectively. Northern blot analysis confirmed that FVIIa binding to TF markedly increased the expression of Cyr61 and CTGF in a time- and dose-dependent manner. FVIIa catalytic activity is required for the gene induction. In addition to FVIIa, thrombin also induced the expression of Cyr61 and CTGF. Hirudin abolished the thrombin-induced expression of these mRNAs but not the FVIIa-induced expression. FVIIa-induced expression of Cyr61 appears not to involve the currently known protease-activated receptors (PARs), whereas thrombin-induced expression involves the activation of PAR1 and possibly an additional PAR. Various intracellular signaling pathway inhibitors exhibited different inhibitory pattern on FVIIa and thrombin-induced up-regulation of Cyr61. Cyr61 and CTGF could act as downstream mediators of FVIIa x TF in affecting various biological processes. PMID- 10799551 TI - Characterization of recombinant phosphatidylinositol 4-kinase beta reveals auto- and heterophosphorylation of the enzyme. AB - Phosphatidylinositol (PI) 4-kinases catalyze the synthesis of PI 4-phosphate, an important intermediate for the synthesis of membrane polyphosphoinositides, regulators of multiple cellular functions. Two mammalian PI 4-kinases have been cloned, a 230-kDa enzyme (alpha-form) and a 110-kDa (beta-form), both of which are inhibited by >0.1 microm concentrations of the PI 3-kinase inhibitor, wortmannin (WT). In the present study, we created a glutathione S-transferase PI4Kbeta fusion protein for expression in Escherichia coli. The purified protein was biologically active and phosphorylated PI in its 4-position with WT sensitivity and kinetic parameters that were identical to those of purified bovine brain PI4Kbeta. In addition to its lipid kinase activity, the enzyme exhibited autophosphorylation that was enhanced by Mn(2+) ions and inhibited by WT and another PI 3-kinase inhibitor, LY 294002. The recombinant protein was unable to transphosphorylate, but its isolated C-terminal catalytic domain still displayed autophosphorylation, suggesting that the autophosphorylation site resides within the C-terminal catalytic domain of the protein and is held in position by intramolecular interactions. Autophosphorylation inhibited subsequent lipid kinase activity, which was reversed upon dephosphorylation, by protein phosphatases, PP1 and PP2A(1), suggesting that it may represent a regulatory mechanism for the enzyme. Phosphorylation of endogenous or overexpressed PI4Kbeta was also observed in COS-7 cells; however, the in vivo phosphorylation of the expressed protein was only partially inhibited by WT and also occurred in a catalytically inactive form of the enzyme, indicating the presence of additional phosphorylation site(s). Successful bacterial expression of PI4Kbeta should aid research on the structure-function relationships of this protein as well as of other, structurally related enzymes. PMID- 10799552 TI - Distinct effects of the UvrD helicase on topoisomerase-quinolone-DNA ternary complexes. AB - Quinolone antibacterial drugs target both DNA gyrase (Gyr) and topoisomerase IV (Topo IV) and form topoisomerase-quinolone-DNA ternary complexes. The formation of ternary complexes results in the inhibition of DNA replication and leads to the generation of double-strand breaks and subsequent cell death. Here, we have studied the consequences of collisions between the UvrD helicase and the ternary complexes formed with either Gyr, Topo IV, or a mutant Gyr, Gyr (A59), which does not wrap the DNA strand around itself. We show (i) that Gyr-norfloxacin (Norf) DNA and Topo IV-Norf-DNA, but not Gyr (A59)-Norf-DNA, ternary complexes inhibit the UvrD-catalyzed strand-displacement activity, (ii) that a single-strand break is generated at small portions of the ternary complexes upon their collisions with UvrD, and (iii) that the majority of Topo IV-Norf-DNA ternary complexes become nonreversible when UvrD collides with the Topo IV-Norf-DNA ternary complexes, whereas the majority of Gyr-Norf-DNA ternary complexes remain reversible after their collision with the UvrD helicase. These results indicated that different DNA repair mechanisms might be involved in the repair of Gyr-Norf DNA and Topo IV-Norf-DNA ternary complexes. PMID- 10799553 TI - Cytochrome P450 CYP79A2 from Arabidopsis thaliana L. Catalyzes the conversion of L-phenylalanine to phenylacetaldoxime in the biosynthesis of benzylglucosinolate. AB - Glucosinolates are natural plant products gaining increasing interest as cancer preventing agents and crop protectants. Similar to cyanogenic glucosides, glucosinolates are derived from amino acids and have aldoximes as intermediates. We report cloning and characterization of cytochrome P450 CYP79A2 involved in aldoxime formation in the glucosinolate-producing Arabidopsis thaliana L. The CYP79A2 cDNA was cloned by polymerase chain reaction, and CYP79A2 was functionally expressed in Escherichia coli. Characterization of the recombinant protein shows that CYP79A2 is an N-hydroxylase converting L-phenylalanine into phenylacetaldoxime, the precursor of benzylglucosinolate. Transgenic A. thaliana constitutively expressing CYP79A2 accumulate high levels of benzylglucosinolate. CYP79A2 expressed in E. coli has a K(m) of 6.7 micromol liter(-1) for L phenylalanine. Neither L-tyrosine, L-tryptophan, L-methionine, nor DL homophenylalanine are metabolized by CYP79A2, indicating that the enzyme has a narrow substrate specificity. CYP79A2 is the first enzyme shown to catalyze the conversion of an amino acid to the aldoxime in the biosynthesis of glucosinolates. Our data provide the first conclusive evidence that evolutionarily conserved cytochromes P450 catalyze this step common for the biosynthetic pathways of glucosinolates and cyanogenic glucosides. This strongly indicates that the biosynthesis of glucosinolates has evolved based on a cyanogenic predisposition. PMID- 10799554 TI - Mutations in the catalytic domain of prohormone convertase 2 result in decreased binding to 7B2 and loss of inhibition with 7B2 C-terminal peptide. AB - Prohormone convertases 1 (PC1) and 2 (PC2) are members of a family of subtilisin like proprotein convertases responsible for proteolytic maturation of a number of different prohormones and proneuropeptides. Although sharing more than 50% homology in their catalytic domains, PC1 and PC2 exhibit differences in substrate specificity and susceptibility to inhibitors. In addition to these differences, PC2, unlike PC1 and other members of the family, specifically binds the neuroendocrine protein 7B2. In order to identify determinants responsible for the specific properties of the PC2 catalytic domain, we compared its primary sequence with that of other PCs. This allowed us to distinguish a PC2-specific sequence at positions 242-248. We constructed two PC2 mutants in which residues 242 and 243 and residues 242-248 were replaced with the corresponding residues of PC1. Studies of in vivo cleavage of proenkephalin, in vivo production of alpha-MSH from proopiomelanocortin, and in vitro cleavage of a PC2-specific artificial substrate by mutant PC2s did not reveal profound alterations. On the other hand, both mutant pro-PC2s exhibited a considerably reduced ability to bind to 21-kDa 7B2. In addition, inhibition of mutant PC2-(242-248) by the potent natural inhibitor 7B2 CT peptide was almost completely abolished. Taken together, our results show that residues 242-248 do not play a significant role in defining the substrate specificity of PC2 but do contribute greatly to binding 7B2 and are critical for inhibition with the 7B2 CT peptide. PMID- 10799555 TI - Specific recognition of parental terminal protein by DNA polymerase for initiation of protein-primed DNA replication. AB - The linear genome of Bacillus subtilis phage phi29 has a protein covalently linked to the 5' ends, called parental terminal protein (TP), and is replicated using a free TP as primer. The initiation of phage phi29 DNA replication requires the formation of a DNA polymerase/TP complex that recognizes the replication origins located at the genome ends. The DNA polymerase catalyzes the formation of the initiation complex TP-dAMP, and elongation proceeds coupled to strand displacement. The same mechanism is used by the related phage Nf. However, DNA polymerase and TP from phi29 do not initiate the replication of Nf TP-DNA. To address the question of the specificity of origin recognition, we took advantage of the initiation reaction enhancement in the presence of Mn(2+), allowing us to detect initiation activity in heterologous systems in which DNA polymerase, TP, and template TP-DNA are not from the same phage. Initiation was selectively stimulated when DNA polymerase and TP-DNA were from the same phage, strongly suggesting that specific recognition of origins is brought through an interaction between DNA polymerase and parental TP. PMID- 10799556 TI - Enhanced DNA binding and activation of transcription factors NF-kappa B and AP-1 by acetaldehyde in HEPG2 cells. AB - Because transcription factors NF-kappaB and activator protein-1 (AP-1) are known to regulate gene expression, we have analyzed the role of acetaldehyde in the activation of NF-kappaB and AP-1 in HepG2 cells. Binding activity and transactivation of NF-kappaB and AP-1 were determined by gel retardation assays and transfection of a luciferase reporter construct controlled by kappaB and AP-1 binding sites, respectively. Acetaldehyde enhanced the DNA binding of NF-kappaB and AP-1 by 1 and 4 h, respectively, increasing the kappaB- and AP-1-dependent luciferase expression. Supershift assays revealed the presence of NF-kappaB heterodimers p65/p50 and p50/p52, whereas nuclear c-Jun levels correlated with the DNA binding of AP-1. The enhanced binding of NF-kappaB to DNA by acetaldehyde in intact cells was accompanied by the proteolytic degradation of IkappaB-alpha. However, the addition of acetaldehyde to cytostolic extracts from untreated Hep G2 cells did not affect the DNA binding of AP-1 but activated the NF-kappaB heterodimer p65/p50 in the absence of IkappaB-alpha degradation. Preincubation of HepG2 cells with protein kinase C inhibitors abolished the enhanced DNA binding of NF-kappaB and AP-1 caused by acetaldehyde. Hence, these findings uncover a previously unrecognized role for acetaldehyde in the activation of NF-kappaB and AP-1, which may be of relevance in the alcohol-induced liver disease. PMID- 10799557 TI - The type 8 adenylyl cyclase is critical for Ca2+ stimulation of cAMP accumulation in mouse parotid acini. AB - Capacitative Ca(2+) entry stimulates cAMP synthesis in mouse parotid acini, suggesting that one of the Ca(2+)-sensitive adenylyl cyclases (AC1 or AC8) may play an important role in the regulation of parotid function (Watson, E. L., Wu, Z., Jacobson, K. L., Storm, D. R., Singh, J. C., and Ott, S. M. (1998) Am. J. Physiol. 274, C557-C565). To evaluate the role of AC1 and AC8 in Ca(2+) stimulation of cAMP synthesis in parotid cells, acini were isolated from AC1 mutant (AC1-KO) and AC8 mutant (AC8-KO) mice and analyzed for Ca(2+) stimulation of intracellular cAMP levels. Although Ca(2+) stimulation of intracellular cAMP levels in acini from AC1-KO mice was indistinguishable from wild type mice, acini from AC8-KO mice showed no Ca(2+)-stimulated cAMP accumulation. This indicates that AC8, but not AC1, plays a major role in coupling Ca(2+) signals to cAMP synthesis in parotid acini. Interestingly, treatment of acini from AC8-KO mice with agents, i.e. carbachol and thapsigargin that increase intracellular Ca(2+), lowered cAMP levels. This decrease was dependent upon Ca(2+) influx and independent of phosphodiesterase activation. Immunoblot analysis revealed that AC5/6 and AC3 are expressed in parotid glands. Inhibition of calmodulin (CaM) kinase II with KN-62, or inclusion of the CaM inhibitor, calmidazolium, did not prevent agonist-induced inhibition of stimulated cAMP accumulation. In vitro studies revealed that Ca(2+), independently of CaM, inhibited isoproterenol stimulated AC. Data suggest that agonist augmentation of stimulated cAMP levels is due to activation of AC8 in mouse parotid acini, and strongly support a role for AC5/6 in the inhibition of stimulated cAMP levels. PMID- 10799558 TI - Human white/murine ABC8 mRNA levels are highly induced in lipid-loaded macrophages. A transcriptional role for specific oxysterols. AB - To identify genes that are transcriptionally activated when human macrophages accumulate excess lipids, we employed the mRNA differential display technique using RNA isolated from human monocyte-macrophages incubated in the absence or presence of acetylated low density lipoprotein and sterols (cholesterol and 25 hydroxycholesterol). These studies identified a mRNA whose levels were highly induced in lipid-loaded macrophages. The mRNA encoded the human White protein, a member of the ATP-binding cassette (ABC) transporter superfamily of proteins. The mRNA levels of ABC8, the murine homolog of the human white gene, were also induced when a murine macrophage cell line, RAW264.7, was incubated with acetylated low density lipoprotein and sterols. Additional studies demonstrated that white/ABC8 mRNA levels were induced by specific oxysterols that included 25 , 20(S)-, and 22(R)-hydroxycholesterol, and by a retinoid X receptor-specific ligand. Furthermore, the oxysterol-mediated induction of ABC8 expression in mouse peritoneal macrophages was dependent on the presence of the nuclear oxysterol receptors, liver X receptors (LXRs). Macrophages derived from mice lacking both LXRalpha and LXRbeta failed to up-regulate the expression of ABC8 following incubation with 22(R)-hydroxycholesterol. Oxysterol-dependent induction of white/ABC8 mRNA was blocked by actinomycin D but not by cycloheximide treatment of cells. We conclude that the white and ABC8 genes are primary response genes that are transcriptionally activated by specific oxysterols and that this induction is mediated by the LXR subfamily of nuclear hormone receptors. These data strongly support the hypothesis that white/ABC8 has a role in cellular sterol homeostasis. PMID- 10799560 TI - The molecular physiology of hepatic nuclear factor 3 in the regulation of gluconeogenesis. AB - Glucocorticoids stimulate gluconeogenesis by increasing the rate of transcription of genes that encode gluconeogenic enzymes such as phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase. Previous studies have shown that hepatic nuclear factor 3 (HNF3) is required as an accessory factor for several glucocorticoid-stimulated genes, including PEPCK. Here, we show that adenovirus mediated expression of an HNF3beta protein with a deleted C-terminal transactivation domain (HNF3betaDeltaC) reduces the glucocorticoid-induced expression of the PEPCK and glucose-6-phosphatase genes in H4IIE hepatoma cells. Furthermore, expression of this truncated HNF3 protein results in a proportionate reduction of glucocorticoid-stimulated glucose production from lactate and pyruvate in these cells. The expression of HNF3betaDeltaN, in which the N terminal transactivation domain is deleted, does not exhibit any of these effects. These results provide direct evidence that members of the HNF3 family are required for proper regulation of hepatic gluconeogenesis. Modulation of the function of the HNF3 family of proteins might be used to reduce the excessive hepatic production of glucose that is an important pathophysiologic feature of diabetes mellitus. PMID- 10799559 TI - Successive expression and activation of NFAT family members during thymocyte differentiation. AB - Differentiation of immature CD4(+)CD8(+) thymocytes to mature CD4(+) or CD8(+) T cells is induced by positive selection and appears to involve calcineurin dependent activation of NFAT, a family of transcription factors. NFATx is predominantly expressed in CD4(+)CD8(+) thymocytes, whereas NFATp and NFATc are expressed at much lower levels in the thymus than in mature T cells. However, how or when each NFAT member is involved in the differentiation pathway is unclear. Using an in vitro model system where isolated CD4(+)CD8(+) thymocytes can survive and differentiate into semi-mature CD4-lineage T cells, we suggest that low calcineurin activity sustained for approximately 20 h is required for cell survival and differentiation. Accordingly, the DNA binding activity of NFAT slowly increased during the stimulation of 20 h to induce the differentiation. NFATx significantly contributed to the early rise, but the late increase was mostly due to NFATc activation. Meanwhile, the expression of NFATx mRNA decreased and that of NFATc mRNA increased. The DNA-binding activity of NFATp was detectable but low throughout the stimulation. NFATp became dominantly active after the semi-mature T cells differentiated into mature and activated CD4 T cells. These findings suggest that NFATx and NFATc successively play roles in T cell development. PMID- 10799561 TI - UDP-N-acetylglucosamine pyrophosphorylase, a key enzyme in encysting Giardia, is allosterically regulated. AB - Giardia synthesizes UDP-GalNAc during cyst wall formation (encystment) via a pathway of inducible enzymes similar to that used to synthesize chitin or peptidoglycan and that includes the UTP-requiring UDP-N-acetylglucosamine pyrophosphorylase. Although it has never been reported as a regulatory enzyme in any system studied to date, kinetic data including Hill plots demonstrate clearly that UDP-N-acetylglucosamine pyrophosphorylase activity, purified from encysting Giardia, is allosterically activated anabolically by physiological levels of glucosamine 6-phosphate (3 microm). Capillary electrophoresis demonstrates that within 24 h after trophozoites are induced to encyst, the level of glucosamine 6 phosphate increases 3-fold over that of non-encysting cells and that by 48 h into encystment the level of glucosamine 6-phosphate has decreased to non-encysting levels or below. UDP-N-acetylglucosamine pyrophosphorylase protein is present constitutively in encysting as well as non-encysting cells. UDP-N acetylglucosamine pyrophosphorylase immunoaffinity purified from encysting and non-encysting cells exhibited the same molecular weight, amino acid composition, and circular dichroism spectra. Moreover, regardless of whether the enzyme came from encysting or non-encysting cells, the change in its circular dichroism spectra and up to a 6-fold increase in its specific activity anabolically were due to its activation with glucosamine 6-phosphate. Thus, the data support the idea that UDP-N-acetylglucosamine pyrophosphorylase is a major regulatory point in amino sugar synthesis in encysting Giardia and that its allosteric anabolic activation may shift the equilibrium of this pathway toward UDP-GalNAc synthesis. PMID- 10799562 TI - Association of p130CAS with phosphatidylinositol-3-OH kinase mediates adenovirus cell entry. AB - The Crk-associated substrate, p130(CAS), has been implicated in the regulation of the actin cytoskeleton following ligation of cell integrins with the extracellular matrix. Integrin-mediated cell adhesion involves p130(CAS) association with focal adhesion kinase (p125(FAK)). Internalization/cell entry of type 2 and type 5 adenoviruses (Ad) is also mediated by alpha(v) integrins. However, expression of dominant negative forms of p125(FAK) does not alter virus entry, and Ad entry occurs normally in p125(FAK)-deficient fibroblasts. We now provide evidence that Ad internalization, a process which is mediated by alpha(v) integrins, also requires p130(CAS) and phosphatidylinositol-3-OH kinase (PI 3 kinase). Ad induces p130(CAS) phosphorylation and inhibition of p130(CAS) phosphorylation by tyrphostin and genistein, or expression of the substrate domain deleted p130(CAS) blocks Ad internalization. p130(CAS) was also found to associate with the p85 subunit of PI 3-kinase through its proline-rich domain during virus internalization and expression of p130(CAS) containing a deleted proline-rich domain (PRD) inhibited adenovirus cell entry. We showed further that the RPLPSPP motif in the proline-rich region of p130(CAS) interacts with the SH3 domain of p85/PI 3-kinase. These studies reveal the molecular basis by which p130(CAS) coordinates the signaling pathways involved in integrin-mediated Ad endocytosis. PMID- 10799563 TI - Beta-cell differentiation factor Nkx6.1 contains distinct DNA binding interference and transcriptional repression domains. AB - beta-Cell differentiation factor Nkx6.1 is a homeodomain protein expressed in developing and mature beta-cells in the pancreatic islets of Langerhans. To understand how it contributes to beta-cell development and function, we characterized its DNA binding and transactivation properties. A single copy of the homeodomain of Nkx6. 1 binds to a strictly conserved 8-base pair DNA consensus sequence, TTAATTAC; even minor variations to this consensus reduce DNA binding affinity significantly. Full-length Nkx6.1, however, has markedly reduced DNA binding affinity due to an acidic domain at the carboxyl end of the molecule that functions as a mobile binding interference domain capable of interrupting the interaction between DNA and DNA binding domains of the helix-turn-helix type. When expressed in fibroblast cell lines, Nkx6.1 represses transcription through isolated Nkx6.1 binding sites; in beta-cell lines, Nkx6.1 specifically represses the intact insulin promoter through TAAT-containing sequences. In Gal4 one-hybrid fusion studies, transcriptional repression maps to a discreet region within the amino terminus. Our findings suggest a model in which Nkx6.1, regulated by interactions through its carboxyl terminus, directs the repression of specific genes in developing and mature beta-cells. PMID- 10799564 TI - Differential effects of sphingomyelin hydrolysis and resynthesis on the activation of NF-kappa B in normal and SV40-transformed human fibroblasts. AB - The precise role of ceramide in NF-kappaB signaling remains unclear. The recent observation of differential sphingomyelin synthase (SMS) activity in normal (low SMS) versus SV40-transformed (high SMS) WI38 human lung fibroblasts provides an opportunity to assess the involvement of ceramide and SMS in NF-kappaB activation. Treatment of normal WI38 fibroblasts with bacterial sphingomyelinase resulted in a 4-fold elevation of ceramide and blocked NF-kappaB activation by serum stimulation. Such inhibition was not observed in SV40-transformed fibroblasts. Under regular growth conditions, after sphingomyelinase was washed out, normal WI38 did not show SM re-synthesis nor NF-kappaB activation. In SV40 WI38, on the other hand, sphingomyelinase washout induced resynthesis of SM due to the action of SMS on ceramide generated at the plasma membrane. NF-kappaB activation correlated with SM resynthesis. This activation was abrogated by D609, which inhibited SM resynthesis but not the initial formation of ceramide. The differential activity of SMS may explain the effects of ceramide in NF-kappaB signaling: in the absence of significant SMS activity, ceramide inhibits NF kappaB, whereas with high SMS, the conversion of the ceramide signal to a diacylglycerol signal by the action of SMS stimulates NF-kappaB. These results also suggest a role for SMS in regulating NF-kappaB. PMID- 10799567 TI - Functional analyses of the EBNA1 origin DNA binding protein of Epstein-Barr virus. AB - The EBNA1 protein of Epstein-Barr virus (EBV) governs the replication and segregation of the viral episomes in latently infected cells and transactivates the expression of other EBV latency proteins through direct interactions with DNA sequences in the EBV latent origin of replication, oriP. To better understand how EBNA1 controls these processes, we have assessed the contribution of various EBNA1 sequences to its replication, segregation, and transactivation functions. Here we show that EBNA1 residues 325 to 376 are responsible for the transactivation activity of EBNA1. This region coincides with the DNA looping domain previously shown to mediate interactions at a distance between DNA-bound EBNA1 molecules. The same residues mediate DNA segregation but have no apparent role in DNA replication, indicating that the replication and transcription activation activities of EBNA1 are distinct. The acidic C-terminal tail of EBNA1 was not found to contribute to replication, transactivation, or segregation. We have also investigated the functional significance of two structural motifs within the DNA binding and dimerization domains of EBNA1, the proline loop and the WF motif. Although the amino acids in these motifs do not directly contact the DNA, both of these motifs were found to contribute to EBNA1 functions by increasing the DNA-binding ability of EBNA1. Mechanisms by which DNA binding is stimulated by these motifs are discussed. PMID- 10799568 TI - The epithelial integrin alphavbeta6 is a receptor for foot-and-mouth disease virus. AB - Field isolates of foot-and-mouth disease virus (FMDV) have been shown to use the RGD-dependent integrin alphavbeta3 as a cellular receptor on cultured cells. However, several other RGD-dependent integrins may have the potential to act as receptors for FMDV in vivo. Of these, alphavbeta6 is a likely candidate for use as a receptor by FMDV as it is expressed on epithelial cells, which correlates with the tissue tropism of the virus. In this report, we show that human colon carcinoma cells (SW480) that are normally nonpermissive for FMDV become susceptible to infection as a result of transfection with the integrin beta6 subunit and expression of alphavbeta6 at the cell surface. Integrin alphavbeta6 is the major site for virus attachment on the beta6-transfected cells, and binding to alphavbeta6 serves to increase the rate of virus entry into these cells. In addition, we show that virus binding and infection of the beta6 transfected cells is mediated through an RGD-dependent interaction that is specifically inhibited by a monoclonal antibody (10D5) that recognizes alphavbeta6. These studies establish a role for alphavbeta6 as a cellular receptor for FMDV. PMID- 10799569 TI - Modulation of Dengue virus infection in human cells by alpha, beta, and gamma interferons. AB - A role for interferon (IFN) in modulating infection by dengue virus (DV) has been suggested by studies in DV-infected patients and IFN receptor-deficient mice. To address how IFN modulates DV type 2 infection, we have assayed IFN-alpha, -beta, and -gamma for the ability to enhance or diminish antibody-independent and antibody-dependent cell infection using a competitive, asymmetric reverse transcriptase-mediated PCR (RT-PCR) assay that quantitates positive and negative strands of viral RNA, a flow cytometric assay that measures viral antigen, and a plaque assay that analyzes virion production. Our data suggest that IFN-alpha and -beta protect cells against DV infection in vitro. Treatment of hepatoma cells with IFN-alpha or -beta decreases viral RNA levels greater than 1, 000-fold, the percentage of cells infected 90 to 95%, and the amount of infectious virus secreted 150- to 100,000-fold. These results have been reproduced with several cell types and viral strains, including low-passage isolates. In contrast, IFN gamma has a more variable effect depending on the cell type and pathway of infection. Quantitative RT-PCR experiments indicate that IFN inhibits DV infection by preventing the accumulation of negative-strand viral RNA. PMID- 10799570 TI - Assembly of the coronavirus envelope: homotypic interactions between the M proteins. AB - The viral membrane proteins M and E are the minimal requirements for the budding of coronavirus particles. Since the E protein occurs in particles only in trace amounts, the lateral interactions between the M proteins apparently generate the major driving force for envelope formation. By using coimmunoprecipitation and envelope incorporation assays, we provide extensive evidence for the existence of such M-M interactions. In addition, we determined which domains of the M protein are involved in this homotypic association, using a mutagenetic approach. Mutant M proteins which were not able to assemble into viruslike particles (VLPs) by themselves (C. A. M. de Haan, L. Kuo, P. S. Masters, H. Vennema, and P. J. M. Rottier, J. Virol. 72:6838-6850, 1998) were tested for the ability to associate with other M proteins and to be rescued into VLPs formed by assembly-competent M proteins. We found that M proteins lacking parts of the transmembrane cluster, of the amphipathic domain, or of the hydrophilic carboxy-terminal tail, or M proteins that had their luminal domain replaced by heterologous ectodomains, were still able to associate with assembly-competent M proteins, resulting in their coincorporation into VLPs. Only a mutant M protein in which all three transmembrane domains had been replaced lost this ability. The results indicate that M protein molecules interact with each other through multiple contact sites, particularly at the transmembrane level. Finally, we tested the stringency with which membrane proteins are selected for incorporation into the coronavirus envelope by probing the coassembly of some foreign proteins. The observed efficient exclusion from budding of the vesicular stomatitis virus G protein and the equine arteritis virus M protein indicates that envelope assembly is indeed a highly selective sorting process. The low but detectable incorporation of CD8 molecules, however, demonstrated that this process is not perfect. PMID- 10799571 TI - Murine cytomegalovirus stimulates cellular thymidylate synthase gene expression in quiescent cells and requires the enzyme for replication. AB - Herpesviruses accomplish DNA replication either by expressing their own deoxyribonucleotide biosynthetic genes or by stimulating the expression of the corresponding cellular genes. Cytomegalovirus (CMV) has adopted the latter strategy to allow efficient replication in quiescent cells. In the present report, we show that murine CMV (MCMV) infection of quiescent fibroblasts induces both mRNA and protein corresponding to the cellular thymidylate synthase (TS) gene, which encodes the enzyme that catalyzes the de novo synthesis of thymidylic acid. The increase in TS gene expression was due to an increase in gene transcription, since the activity of a reporter gene driven by the mouse TS promoter was induced following MCMV infection. Mutagenesis of the potential E2F responsive element immediately upstream from the TS essential promoter region abolished the virus-mediated stimulation of the TS promoter, suggesting that the transactivating activity of MCMV infection was E2F dependent. Cotransfection experiments revealed that expression of the viral immediate-early 1 protein was sufficient to mediate the increase in TS promoter activity. Finally, MCMV replication and viral DNA synthesis were found to be inhibited by ZD1694, a quinazoline-based folate analog that inhibits TS activity. These results demonstrate that upregulation of cellular TS expression is required for efficient MCMV replication in quiescent cells. PMID- 10799572 TI - Synergistic action of GA-binding protein and glucocorticoid receptor in transcription from the mouse mammary tumor virus promoter. AB - B lymphocytes are among the first cells to be infected by mouse mammary tumor virus (MMTV), and they play a crucial role in its life cycle. To study transcriptional regulation of MMTV in B cells, we have analyzed two areas of the long terminal repeat (LTR) next to the glucocorticoid receptor binding site, fp1 (at position -139 to -146 from the cap site) and fp2 (at -157 to -164). Both showed B-cell-specific protection in DNase I in vitro footprinting assays and contain binding sites for Ets transcription factors, a large family of proteins involved in cell proliferation and differentiation and oncogenic transformation. In gel retardation assays, fp1 and fp2 bound the heterodimeric Ets factor GA binding protein (GABP) present in B-cell nuclear extracts, which was identified by various criteria: formation of dimers and tetramers, sensitivity to pro oxidant conditions, inhibition of binding by specific antisera, and comigration of complexes with those formed by recombinant GABP. Mutations which prevented complex formation in vitro abolished glucocorticoid-stimulated transcription from an MMTV LTR linked to a reporter gene in transiently transfected B-cell lines, whereas they did not affect the basal level. Exogenously expressed GABP resulted in an increased level of hormone response of the LTR reporter plasmid and produced a synergistic effect with the coexpressed glucocorticoid receptor, indicating cooperation between the two. This is the first example of GABP cooperation with a steroid receptor, providing the opportunity for studying the integration of their intracellular signaling pathways. PMID- 10799573 TI - Induction of CD4(+) T-cell-independent immunoglobulin responses by inactivated influenza virus. AB - Through cognate interaction between antigen-specific B-cell and CD4(+) alphabeta T cells, the CD4(+) alphabeta T cells secrete cytokines that initiate immunoglobulin (Ig) class switching from IgM to IgG. In this study, we show that formalin-inactivated influenza PR8 virus induces virus-specific IgM and IgG responses in the absence of CD4(+) T cells and that all four subclasses of IgG are produced. The immunized CD4-deficient mice were also found to be completely protected against lethal infection with live, pathogenic influenza virus. The ability of CD4(+) T-cell-deficient mice to generate these IgG responses was not found to be impaired when these mice were depleted of CD8(+) T cells with an anti CD8 monoclonal antibody. In contrast, alphabeta T-cell-deficient mice (TCRbeta(-/ )) were not found to produce significant amounts of IgG upon immunization with formalin-inactivated PR8 virus. These results suggest that CD4(-) CD8(-) double negative alphabeta T cells are playing a role in regulating Ig class switching in the absence of CD4(+) T cells. PMID- 10799574 TI - Pegylated alpha interferon is an effective treatment for virulent venezuelan equine encephalitis virus and has profound effects on the host immune response to infection. AB - Venezuelan equine encephalitis virus (VEEV) is a highly infectious alphavirus endemic in parts of Central and South America. The disease is transmitted by mosquitoes, and the natural reservoir is the small rodent population, with epidemics occurring in horses and occasionally humans. Following infection, VEEV replicates in lymphoid tissues prior to invasion of the central nervous system. Treatment of VEEV-infected BALB/c mice with polyethylene glycol-conjugated alpha interferon (PEG IFN-alpha) results in a greatly enhanced survival from either a subcutaneous or an aerosol infection. Virus is undetectable within PEG IFN-alpha treated individuals by day 30 postinfection (p.i.). Treatment results in a number of changes to the immune response characteristics normally associated with VEEV infection. Increased macrophage activation occurs in PEG IFN-alpha-treated BALB/c mice infected with VEEV. The rapid activation of splenic CD4, CD8, and B cells by day 2 p.i. normally associated with VEEV infection is absent in PEG IFN-alpha treated mice. The high tumor necrosis factor alpha production by macrophages from untreated mice is greatly diminished in PEG IFN-alpha-treated mice. These results suggest key immunological mechanisms targeted by this lethal alphavirus that can be modulated by prolonged exposure to IFN-alpha. PMID- 10799575 TI - Coreceptor competition for association with CD4 may change the susceptibility of human cells to infection with T-tropic and macrophagetropic isolates of human immunodeficiency virus type 1. AB - The chemokine receptors CCR5 and CXCR4 were found to function in vivo as the principal coreceptors for M-tropic and T-tropic human immunodeficiency virus (HIV) strains, respectively. Since many primary cells express multiple chemokine receptors, it was important to determine if the efficiency of virus-cell fusion is influenced not only by the presence of the appropriate coreceptor (CXCR4 or CCR5) but also by the levels of other coreceptors expressed by the same target cells. We found that in cells with low to medium surface CD4 density, coexpression of CCR5 and CXCR4 resulted in a significant reduction in the fusion with CXCR4 domain (X4) envelope-expressing cells and in their susceptibility to infection with X4 viruses. The inhibition could be reversed either by increasing the density of surface CD4 or by antibodies against the N terminus and second extracellular domains of CCR5. In addition, treatment of macrophages with a combination of anti-CCR5 antibodies or beta-chemokines increased their fusion with X4 envelope-expressing cells. Conversely, overexpression of CXCR4 compared with CCR5 inhibited CCR5-dependent HIV-dependent fusion in 3T3.CD4.401 cells. Thus, coreceptor competition for association with CD4 may occur in vivo and is likely to have important implications for the course of HIV type 1 infection, as well as for the outcome of coreceptor-targeted therapies. PMID- 10799576 TI - Antiapoptotic herpesvirus Bcl-2 homologs escape caspase-mediated conversion to proapoptotic proteins. AB - The antiapoptotic Bcl-2 and Bcl-x(L) proteins of mammals are converted into potent proapoptotic factors when they are cleaved by caspases, a family of apoptosis-inducing proteases (E. H.-Y. Cheng, D. G. Kirsch, R. J. Clem, R. Ravi, M. B. Kastan, A. Bedi, K. Ueno, and J. M. Hardwick, Science 278:1966-1968, 1997; R. J. Clem, E. H.-Y. Cheng, C. L. Karp, D. G. Kirsch, K. Ueno, A. Takahashi, M. B. Kastan, D. E. Griffin, W. C. Earnshaw, M. A. Veliuona, and J. M. Hardwick, Proc. Natl. Acad. Sci. USA 95:554-559, 1998). Gamma herpesviruses also encode homologs of the Bcl-2 family. All tested herpesvirus Bcl-2 homologs possess antiapoptotic activity, including the more distantly related homologs encoded by murine gammaherpesvirus 68 (gammaHV68) and bovine herpesvirus 4 (BHV4), as described here. To determine if viral Bcl-2 proteins can be converted into death factors, similar to their cellular counterparts, five herpesvirus Bcl-2 homologs from five different viruses were tested for their susceptibility to caspases. Only the viral Bcl-2 protein encoded by gammaHV68 was susceptible to caspase digestion. However, unlike the caspase cleavage products of cellular Bcl-2, Bcl x(L), and Bid, which are potent inducers of apoptosis, the cleavage product of gammaHV68 Bcl-2 lacked proapoptotic activity. KSBcl-2, encoded by the Kaposi's sarcoma-associated herpesvirus, was the only viral Bcl-2 homolog that was capable of killing cells when expressed as an N-terminal truncation. However, because KSBcl-2 was not cleavable by caspases, the latent proapoptotic activity of KSBcl 2 apparently cannot be released. The Bcl-2 homologs encoded by herpesvirus saimiri, Epstein-Barr virus, and BHV4 were not cleaved by apoptotic cell extracts and did not possess latent proapoptotic activities. Thus, herpesvirus Bcl-2 homologs escape negative regulation by retaining their antiapoptotic activities and/or failing to be converted into proapoptotic proteins by caspases during programmed cell death. PMID- 10799577 TI - The hepatitis B virus core promoter is strongly activated by the liver nuclear receptor fetoprotein transcription factor or by ectopically expressed steroidogenic factor 1. AB - Orphan nuclear receptor fetoprotein transcription factor (FTF) was previously identified as a specific regulator of the alpha(1)-fetoprotein gene during early liver development and in response to hormonal signals (L. Galarneau, J.-F. Pare, D. Allard, D. Hamel, L. Levesque, J. D. Tugwood, S. Green, and L. Belanger, Mol. Cell. Biol. 16:3853-3865, 1996). Here we report a functional analysis of FTF interactions with the hepatitis B virus (HBV) nucleocapsid promoter. DNA-protein binding assays show that the HBV core promoter contains two high-affinity FTF binding sites and a third, lower-affinity site shared with other receptors. Transfections in HepG2, Hep3B, and PLC/PRF/5 hepatoma cells using chloramphenicol acetyltransferase reporter genes with the nucleocapsid promoter linked or not linked to enhancer I indicate that FTF is a potent activator of the HBV core promoter, more efficient than HNF4alpha, HNF3alpha, HNF3beta, or C/EBPalpha. Steroidogenic factor 1, a close FTF homolog which binds to the same DNA motif and is expressed ectopically in HepG2 cells, seems to be an even stronger inducer than FTF. Point mutations of the FTF-binding sites indicate direct FTF activatory effects on the core promoter and the use of both high-affinity sites for productive interaction between the core promoter and enhancer I. Coexpression assays further indicate that FTF and HNF4alpha are the most efficient partners for coactivation of the pregenomic core promoter, which may largely account for the hepatic tropism and the early amplification of HBV infection. Carboxy terminus-truncated FTF behaves as a dominant negative mutant to compete all three FTF sites and strongly deactivate core promoter interactions with enhancer I; this suggests possible new ways to interfere with HBV infection. PMID- 10799578 TI - Loss of G(1)/S checkpoint in human immunodeficiency virus type 1-infected cells is associated with a lack of cyclin-dependent kinase inhibitor p21/Waf1. AB - Productive high-titer infection by human immunodeficiency virus type 1 (HIV-1) requires the activation of target cells. Infection of quiescent peripheral CD4 lymphocytes by HIV-1 results in incomplete, labile reverse transcripts and lack of viral progeny formation. An interplay between Tat and p53 has previously been reported, where Tat inhibited the transcription of the p53 gene, which may aid in the development of AIDS-related malignancies, and p53 expression inhibited HIV-1 long terminal repeat transcription. Here, by using a well-defined and characterized stress signal, gamma irradiation, we find that upon gamma irradiation, HIV-1-infected cells lose their G(1)/S checkpoints, enter the S phase inappropriately, and eventually apoptose. The loss of the G(1)/S checkpoint is associated with a loss of p21/Waf1 protein and increased activity of a major G(1)/S kinase, namely, cyclin E/cdk2. The p21/Waf1 protein, a known cyclin dependent kinase inhibitor, interacts with the cdk2/cyclin E complex and inhibits progression of cells into S phase. We find that loss of the G(1)/S checkpoint in HIV-1-infected cells may in part be due to Tat's ability to bind p53 (a known activator of the p21/Waf1 promoter) and sequester its transactivation activity, as seen in both in vivo and in vitro transcription assays. The loss of p21/Waf1 in HIV-1-infected cells was specific to p21/Waf1 and did not occur with other KIP family members, such as p27 (KIP1) and p57 (KIP2). Finally, the advantage of a loss of the G(1)/S checkpoint for HIV-1 per se may be that it pushes the host cell into the S phase, which may then allow subsequent virus-associated processes, such as RNA splicing, transport, translation, and packaging of virion specific genes, to occur. PMID- 10799579 TI - Host protein interactions with the 3' end of bovine coronavirus RNA and the requirement of the poly(A) tail for coronavirus defective genome replication. AB - RNA viruses have 5' and 3' untranslated regions (UTRs) that contain specific signals for RNA synthesis. The coronavirus genome is capped at the 5' end and has a 3' UTR that consists of 300 to 500 nucleotides (nt) plus a poly(A) tail. To further our understanding of coronavirus replication, we have begun to examine the involvement of host factors in this process for two group II viruses, bovine coronavirus (BCV) and mouse hepatitis coronavirus (MHV). Specific host protein interactions with the BCV 3' UTR [287 nt plus poly(A) tail] were identified using gel mobility shift assays. Competition with the MHV 3' UTR [301 nt plus poly(A) tail] suggests that the interactions are conserved for the two viruses. Proteins with molecular masses of 99, 95, and 73 kDa were detected in UV cross-linking experiments. Less heavily labeled proteins were also detected in the ranges of 40 to 50 and 30 kDa. The poly(A) tail was required for binding of the 73-kDa protein. Immunoprecipitation of UV-cross-linked proteins identified the 73-kDa protein as the cytoplasmic poly(A)-binding protein (PABP). Replication of the defective genomes BCV Drep and MHV MIDI-C, along with several mutants, was used to determine the importance of the poly(A) tail. Defective genomes with shortened poly(A) tails consisting of 5 or 10 A residues were replicated after transfection into helper virus-infected cells. BCV Drep RNA that lacked a poly(A) tail did not replicate, whereas replication of MHV MIDI-C RNA with a deleted tail was detected after several virus passages. All mutants exhibited delayed kinetics of replication. Detectable extension or addition of the poly(A) tail to the mutants correlated with the appearance of these RNAs in the replication assay. RNAs with shortened poly(A) tails exhibited less in vitro PABP binding, suggesting that decreased interactions with the protein may affect RNA replication. The data strongly indicate that the poly(A) tail is an important cis-acting signal for coronavirus replication. PMID- 10799580 TI - Cell fusion activity of hepatitis C virus envelope proteins. AB - To examine the cell fusion activity of hepatitis C virus (HCV) envelope proteins (E1 and E2), we have established a sensitive cell fusion assay based on the activation of a reporter gene as described previously (O. Nussbaum, C. C. Broder, and E. A. Berger, J. Virol. 68:5411-5422, 1994). The chimeric HCV E1 and E2 proteins, each consisting of the ectodomain of the E1 and E2 envelope protein and the transmembrane and cytoplasmic domains of the vesicular stomatitis virus G glycoprotein, were expressed on the cell surface. Cells expressing the chimeric envelope proteins and T7 RNA polymerase were cocultured with the various target cell lines transfected with a reporter plasmid encoding the luciferase gene under the control of the T7 promoter. After cocultivation, the cell fusion activity was determined by the expression of luciferase in the cocultured cells. The induction of cell fusion requires both the chimeric E1 and E2 proteins and occurs in a low pH-dependent manner. Although it has been shown that HCV E2 protein binds human CD81 (P. Pileri, Y. Uematsu, S. Campagnoli, G. Galli, F. Falugi, R. Petracca, A. J. Weiner, M. Houghton, D. Rosa, G. Grandi, and S. Abrignani, Science 282:938 941, 1998), the expression of human CD81 alone is not sufficient to confer susceptibility to cell fusion in the mouse cell line. Treatment of the target cells with pronase, heparinase, or heparitinase reduced the cell fusion activity induced by the chimeric envelope proteins. These results suggest (i) that both HCV E1 and E2 proteins are responsible for fusion with the endosomal membrane after endocytosis and (ii) that certain protein molecules other than human CD81 and some glycosaminoglycans on the cell surface are also involved in the cell fusion induced by HCV. PMID- 10799581 TI - Simian immunodeficiency virus utilizes human and sooty mangabey but not rhesus macaque STRL33 for efficient entry. AB - It has been established that many simian immunodeficiency virus (SIV) isolates utilize the orphan receptors GPR15 and STRL33 about as efficiently as the chemokine receptor CCR5 for entry into target cells. Most studies were performed, however, with coreceptors of human origin. We found that SIV from captive rhesus macaques (SIVmac) can utilize both human and simian CCR5 and GPR15 with comparable efficiencies. Strikingly, however, only human STRL33 (huSTRL33), not rhesus macaque STRL33 (rhSTRL33), functioned efficiently as an entry cofactor for a variety of isolates of SIVmac and SIV from sooty mangabeys. A single amino acid substitution of S30R in huSTRL33 impaired coreceptor activity, and the reverse change in rhSTRL33 greatly increased coreceptor activity. In comparison, species specific sequence variations in N-terminal tyrosines in STRL33 had only moderate effects on SIV entry. These results show that a serine residue located just outside of the cellular membrane in the N terminus of STRL33 is critical for SIV coreceptor function. Interestingly, STRL33 derived from sooty mangabeys, a natural host of SIV, also contained a serine at the corresponding position and was used efficiently as an entry cofactor. These results suggest that STRL33 is not a relevant coreceptor in the SIV/macaque model but may play a role in SIV replication and transmission in naturally infected sooty mangabeys. PMID- 10799582 TI - Pseudorabies virus glycoprotein K requires the UL20 gene product for processing. AB - Glycoprotein K (gK) of pseudorabies virus (PrV) has recently been identified as a virion component which is dispensable for viral entry but required for direct cell-to-cell spread. Electron microscopic data suggested a possible function of gK in virus egress by preventing immediate fusion of released virus particles with the plasma membrane (B. G. Klupp, J. Baumeister, P. Dietz, H. Granzow, and T. C. Mettenleiter, J. Virol. 72:1949-1958, 1998). For more detailed analysis, a PrV mutant with a deletion of the UL53 (gK) open reading frame (ORF) from codons 48 to 275 was constructed, and the protein was analyzed with two monoclonal antibodies directed against PrV gK. The salient findings of this report are as follows. (i) From the PrV UL53 ORF, a functional gK is translated only from the first in-frame methionine. From the second in-frame methionine, a nonfunctional product is expressed which is not incorporated into virions. (ii) When constitutively expressed in a stable cell line without other viral proteins, gK is only incompletely processed. After superinfection with gK-deletion mutants, proper processing is restored and mature gK is incorporated into virions. (iii) The UL20 gene product is specifically required for processing of gK. gK is not correctly processed in a UL20 deletion mutant of PrV, and superinfection of gK expressing cells with PrV-UL20(-) does not restore processing. However, all other known structural viral glycoproteins appear to be processed normally in PrV-UL20( )-infected cells. (iv) Coexpression of gK and UL20 restored gK processing at least partially. Thus, our data show that the UL20 gene product is required for proper processing of PrV gK. PMID- 10799583 TI - Variable-loop-deleted variants of the human immunodeficiency virus type 1 envelope glycoprotein can be stabilized by an intermolecular disulfide bond between the gp120 and gp41 subunits. AB - We have described an oligomeric gp140 envelope glycoprotein from human immunodeficiency virus type 1 that is stabilized by an intermolecular disulfide bond between gp120 and the gp41 ectodomain, termed SOS gp140 (J. M. Binley, R. W. Sanders, B. Clas, N. Schuelke, A. Master, Y. Guo, F. Kajumo, D. J. Anselma, P. J. Maddon, W. C. Olson, and J. P. Moore, J. Virol. 74:627-643, 2000). In this protein, the protease cleavage site between gp120 and gp41 is fully utilized. Here we report the characterization of gp140 variants that have deletions in the first, second, and/or third variable loop (V1, V2, and V3 loops). The SOS disulfide bond formed efficiently in gp140s containing a single loop deletion or a combination deletion of the V1 and V2 loops. However, deletion of all three variable loops prevented formation of the SOS disulfide bond. Some variable-loop deleted gp140s were not fully processed to their gp120 and gp41 constituents even when the furin protease was cotransfected. The exposure of the gp120-gp41 cleavage site is probably affected in these proteins, even though the disabling change is in a region of gp120 distal from the cleavage site. Antigenic characterization of the variable-loop-deleted SOS gp140 proteins revealed that deletion of the variable loops uncovers cryptic, conserved neutralization epitopes near the coreceptor-binding site on gp120. These modified, disulfide stabilized glycoproteins might be useful as immunogens. PMID- 10799584 TI - A single amino acid change in the Newcastle disease virus fusion protein alters the requirement for HN protein in fusion. AB - The role of a leucine heptad repeat motif between amino acids 268 and 289 in the structure and function of the Newcastle disease virus (NDV) F protein was explored by introducing single point mutations into the F gene cDNA. The mutations affected either folding of the protein or the fusion activity of the protein. Two mutations, L275A and L282A, likely interfered with folding of the molecule since these proteins were not proteolytically cleaved, were minimally expressed at the cell surface, and formed aggregates. L268A mutant protein was cleaved and expressed at the cell surface although the protein migrated slightly slower than wild type on polyacrylamide gels, suggesting an alteration in conformation or processing. L268A protein was fusion inactive in the presence or absence of HN protein expression. Mutant L289A protein was expressed at the cell surface and proteolytically cleaved at better than wild-type levels. Most importantly, this protein mediated syncytium formation in the absence of HN protein expression although HN protein enhanced fusion activity. These results show that a single amino acid change in the F(1) portion of the NDV F protein can alter the stringent requirement for HN protein expression in syncytium formation. PMID- 10799586 TI - A metastable form of the large envelope protein of duck hepatitis B virus: low-pH release results in a transition to a hydrophobic, potentially fusogenic conformation. AB - We have examined the structure and fusion potential of the duck hepatitis B virus (DHBV) envelope proteins by treating subviral particles with deforming agents known to release envelope proteins of viruses from a metastable to a fusion active state. Exposure of DHBV particles to low pH triggered a major structural change in the large envelope protein (L), resulting in exposure of trypsin sites within its S domain but without affecting the same region in the small surface protein (S) subunits. This conformational change was associated with increased hydrophobicity of the particle surface, most likely arising from surface exposure of the hydrophobic first transmembrane domain (TM1). In the hydrophobic conformation, DHBV particles were able to bind to liposomes and intact cells, while in their absence these particles aggregated, resulting in viral inactivation. These results suggests that some L molecules are in a spring-loaded metastable state which, when released, exposes a previously hidden hydrophobic domain, a transition potentially representing the fusion-active state of the envelope. PMID- 10799585 TI - Mov34 protein from mouse brain interacts with the 3' noncoding region of Japanese encephalitis virus. AB - The plus-sense RNA genome of Japanese encephalitis virus (JEV) contains noncoding regions (NCRs) of 95 and 585 bases at its 5' and 3' ends, respectively. The last 83 nucleotides of the 3'-NCR are predicted to form stable stem-loop (SL) structures. The shape of this 3'-SL structure is highly conserved among divergent flaviviruses even though only small stretches of nucleotide sequence contained within these structures are conserved. These SL structures have been predicted to function as cis-acting signals for RNA replication and as such may bind to viral and cellular proteins that may be involved in viral replication. We have studied the interaction of the JEV 3'-NCR RNA with host proteins using gel retardation assays. We show that the JEV 3'-SL structure RNA forms three complexes with proteins from the S100 cytoplasmic extract prepared from the neonatal mouse brain. These complexes could be obtained in the presence of 200 mM KCl, indicating that the RNA-protein interaction may be physiologically relevant. UV induced cross-linking and Northwestern blotting analyses detected three proteins with apparent molecular masses of 32, 35, and 50 kDa that bound to the JEV 3'-SL structure RNA. Screening of the neonatal mouse brain cDNA library with the JEV 3' SL structure RNA identified a 36-kDa Mov34 protein interacting with it. Competition experiments using the RNA extracted from JEV virions established that the 36-kDa Mov34 protein indeed bound to the JEV genome. Murine Mov34 belongs to a family of proteins whose members have been shown to be involved in RNA transcription and translation. It is, therefore, likely that the murine Mov34 interaction with JEV 3'-NCR has a role in RNA replication. PMID- 10799587 TI - Characterization and construction of functional cDNA clones of Pariacoto virus, the first Alphanodavirus isolated outside Australasia. AB - Pariacoto virus (PaV) was recently isolated in Peru from the Southern armyworm (Spodoptera eridania). PaV particles are isometric, nonenveloped, and about 30 nm in diameter. The virus has a bipartite RNA genome and a single major capsid protein with a molecular mass of 39.0 kDa, features that support its classification as a Nodavirus. As such, PaV is the first Alphanodavirus to have been isolated from outside Australasia. Here we report that PaV replicates in wax moth larvae and that PaV genomic RNAs replicate when transfected into cultured baby hamster kidney cells. The complete nucleotide sequences of both segments of the bipartite RNA genome were determined. The larger genome segment, RNA1, is 3,011 nucleotides long and contains a 973-amino-acid open reading frame (ORF) encoding protein A, the viral contribution to the RNA replicase. During replication, a 414-nucleotide long subgenomic RNA (RNA3) is synthesized which is coterminal with the 3' end of RNA1. RNA3 contains a small ORF which could encode a protein of 90 amino acids similar to the B2 protein of other alphanodaviruses. RNA2 contains 1,311 nucleotides and encodes the 401 amino acids of the capsid protein precursor alpha. The amino acid sequences of the PaV capsid protein and the replicase subunit share 41 and 26% identity with homologous proteins of Flock house virus, the best characterized of the alphanodaviruses. These and other sequence comparisons indicate that PaV is evolutionarily the most distant of the alphanodaviruses described to date, consistent with its novel geographic origin. Although the PaV capsid precursor is cleaved into the two mature capsid proteins beta and gamma, the amino acid sequence at the cleavage site, which is Asn/Ala in all other alphanodaviruses, is Asn/Ser in PaV. To facilitate the investigation of PaV replication in cultured cells, we constructed plasmids that transcribed full length PaV RNAs with authentic 5' and 3' termini. Transcription of these plasmids in cells recreated the replication of PaV RNA1 and RNA2, synthesis of subgenomic RNA3, and translation of viral proteins A and alpha. PMID- 10799588 TI - Mutational analysis of the rubella virus nonstructural polyprotein and its cleavage products in virus replication and RNA synthesis. AB - Rubella virus nonstructural proteins, translated from input genomic RNA as a p200 polyprotein and subsequently processed into p150 and p90 by an intrinsic papain like thiol protease, are responsible for virus replication. To examine the effect of p200 processing on virus replication and to study the roles of nonstructural proteins in viral RNA synthesis, we introduced into a rubella virus infectious cDNA clone a panel of mutations that had variable defective effects on p200 processing. The virus yield and viral RNA synthesis of these mutants were examined. Mutations that completely abolished (C1152S and G1301S) or largely abolished (G1301A) cleavage of p200 resulted in noninfectious virus. Mutations that partially impaired cleavage of p200 (R1299A and G1300A) decreased virus replication. An RNase protection assay revealed that all of the mutants synthesized negative-strand RNA as efficiently as the wild type does but produced lower levels of positive-strand RNA. Our results demonstrated that processing of rubella virus nonstructural protein is crucial for virus replication and that uncleaved p200 could function in negative-strand RNA synthesis, whereas the cleavage products p150 and p90 are required for efficient positive-strand RNA synthesis. PMID- 10799589 TI - Relationship between human immunodeficiency virus type 1 Gag multimerization and membrane binding. AB - The human immunodeficiency virus type 1 (HIV-1) Gag precursor, Pr55(Gag), is necessary and sufficient for the assembly and release of viruslike particles. Binding of Gag to membrane and Gag multimerization are both essential steps in virus assembly, yet the domains responsible for these events have not been fully defined. In addition, the relationship between membrane binding and Gag-Gag interaction remains to be elucidated. To investigate these issues, we analyzed, in vivo, the membrane-binding and assembly properties of a series of C-terminally truncated Gag mutants. Pr55(Gag) was truncated at the C terminus of matrix (MAstop), between the N- and C-terminal domains of capsid (CA146stop), at the C terminus of capsid (p41stop), at the C terminus of p2 (p43stop), and after the N terminal 35 amino acids of nucleocapsid (NC35stop). The ability of these truncated Gag molecules to assemble and release viruslike particles and their capacity to copackage into particles when coexpressed with full-length Gag were determined. We demonstrate that the amount of truncated Gag incorporated into particles is incrementally increased by extension from CA146 to NC35, suggesting that multiple sites in this region are involved in Gag multimerization. Using membrane flotation centrifugation, we observe that MA shows significantly reduced membrane binding relative to full-length Gag but that CA146 displays steady-state membrane-binding properties comparable to those of Pr55(Gag). The finding that the CA146 mutant, which contains only matrix and the N-terminal domain of capsid, exhibits levels of steady-state membrane binding equivalent to those of full length Gag indicates that strong Gag-Gag interaction domains are not required for the efficient binding of HIV-1 Gag to membrane. PMID- 10799590 TI - Epstein-barr virus nuclear antigen 3C activates the latent membrane protein 1 promoter in the presence of Epstein-Barr virus nuclear antigen 2 through sequences encompassing an spi-1/Spi-B binding site. AB - The Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA-3C) protein is a transcriptional regulator of viral and cellular genes that is essential for EBV mediated immortalization of B lymphocytes in vitro. EBNA-3C can inhibit transcription through an association with the cellular DNA-binding protein Jkappa, a function shared by EBNA-3A and EBNA-3B. Here, we report a mechanism by which EBNA-3C can activate transcription from the EBV latent membrane protein 1 (LMP-1) promoter in conjunction with EBNA-2. Jkappa DNA-binding sites were not required for this activation, and a mutant EBNA-3C protein unable to bind Jkappa activated transcription as efficiently as wild-type EBNA-3C, indicating that EBNA 3C can regulate transcription through a mechanism that is independent of Jkappa. Furthermore, activation of the LMP-1 promoter is a unique function of EBNA-3C, not shared by EBNA-3A and EBNA-3B. The DNA element through which EBNA-3C activates the LMP-1 promoter includes a Spi-1/Spi-B binding site, previously characterized as an important EBNA-2 response element. Although this element has considerable homology to mouse immunoglobulin light chain promoter sequences to which the mouse homologue of Spi-1 binds with its dimerization partner IRF4, we demonstrate that the IRF4-like binding sites in the LMP-1 promoter do not play a role in EBNA-3C-mediated activation. Both EBNA-2 and EBNA-3C were required for transcription mediated through a 41-bp region of the LMP-1 promoter encompassing the Spi binding site. However, EBNA-3C had no effect on transcription mediated in conjunction with the EBNA-2 activation domain fused to the GAL4 DNA-binding domain, suggesting that it does not function as an adapter between EBNA-2 and the cellular transcriptional machinery. Like EBNA-2, EBNA-3C bound directly to both Spi-1 and Spi-B in vitro. This interaction was mediated by a region of EBNA-3C encompassing a likely basic leucine zipper (bZIP) domain and the ets domain of Spi-1 or Spi-B, reminiscent of interactions between bZIP and ets domains of other transcription factors that result in their targeting to DNA. There are many examples of regulation of the hematopoietic-specific Spi transcription factors through protein-protein interactions, and a similar regulation by EBNA-3C, in conjunction with EBNA-2, is likely to be an important and unique contribution of EBNA-3C to EBV-mediated immortalization. PMID- 10799591 TI - Circular double-stranded forms of TT virus DNA in the liver. AB - TT virus (TTV) is an unenveloped, circular, and single-stranded DNA virus commonly infecting human beings worldwide. TTV DNAs in paired serum and liver tissues from three viremic individuals were separated by gel electrophoresis and characterized biophysically. TTV DNAs in sera migrated in sizes ranging from 2.0 to 2.5 kb. TTV DNAs in liver tissues, however, migrated at 2.0 to 2.5 kb as well as at 3.5 to 6.1 kb. Both faster- and slower-migrating forms of TTV DNAs in the liver were found to be circular and of the full genomic length of 3.8 kb. TTV DNAs migrating at 2.0 to 2.5 kb, from either serum or liver tissues, were sensitive to S1 nuclease but resistant to restriction endonucleases, and therefore, they were single-stranded. By contrast, TTV DNAs in liver tissues that migrated at 3.5 to 6.1 kb were resistant to S1 nuclease. They migrated at 3.7 to 4.0 kb after digestion with EcoRI, which suggests that they represent circular, double-stranded replicative intermediates of TTV. When TTV DNAs were subjected to strand-specific primer extension and then amplified by PCR with internal primers, those in serum were found to be minus-stranded DNAs while those in liver tissues were found to be a mixture of plus- and minus-stranded DNAs. These results suggest that TTV replicates in the liver via a circular double-stranded DNA. PMID- 10799592 TI - Two distinct activities contribute to the oncogenic potential of the adenovirus type 5 E4orf6 protein. AB - Previous studies have shown that the adenovirus type 5 (Ad5) E4orf6 gene product displays features of a viral oncoprotein. It initiates focal transformation of primary rat cells in cooperation with Ad5 E1 genes and confers multiple additional transformed properties on E1-expressing cells, including profound morphological alterations and dramatically accelerated tumor growth in nude mice. It has been reported that E4orf6 binds to p53 and, in the presence of the Ad5 E1B 55kDa protein, antagonizes p53 stability by targeting the tumor suppressor protein for active degradation. In the present study, we performed a comprehensive mutant analysis to assign transforming functions of E4orf6 to distinct regions within the viral polypeptide and to analyze a possible correlation between E4orf6-dependent p53 degradation and oncogenesis. Our results show that p53 destabilization maps to multiple regions within both amino- and carboxy-terminal parts of the viral protein and widely cosegregates with E4orf6 dependent acceleration of tumor growth, indicating that both effects are related. In contrast, promotion of focus formation and morphological transformation require only a carboxy-terminal segment of the E4 protein. Thus, these effects are completely independent of p53 stability, but may involve other interactions with the tumor suppressor. Our results demonstrate that at least two distinct activities contribute to the oncogenic potential of Ad5 E4orf6. Although genetically separable, both activities are largely mediated through a novel highly conserved, cysteine-rich motif and a recently described arginine-faced amphipathic alpha helix, which resides within a carboxy-terminal "oncodomain" of the viral protein. PMID- 10799593 TI - Differential activity of two non-hr origins during replication of the baculovirus Autographa californica nuclear polyhedrosis virus genome. AB - The identification of potential baculovirus origins of replication (ori) has involved the generation and characterization of defective interfering particles that contain major genomic deletions yet retain their capability to replicate by testing the replication ability of transiently transfected plasmids carrying viral sequences in infected cells. So far, there has not been any evidence to demonstrate the actual utilization of these putative origins in Autographa californica multinucleocapsid nuclear polyhedrosis virus (AcMNPV) replication. By using the method of origin mapping by competitive PCR, we have obtained quantitative data for the ori activity of the HindIII-K region and the ie-1 promoter sequence in AcMNPV. We also provide evidence for differential activity of the two ori in the context of the viral genome through the replication phase of viral infection. Comparison of the number of molecules representing the HindIII-K and ie-1 origins vis-a-vis the non-ori polH region in a size-selected nascent DNA preparation revealed that the HindIII-K ori is utilized approximately 14 times more efficiently than the ie-1 region during the late phase of infection. HindIII-K also remains the more active ori through the early and middle replication phases. Our results provide in vivo evidence in support of the view that AcMNPV replication involves multiple ori that are activated with vastly different efficiencies during the viral infection cycle. PMID- 10799594 TI - Involvement of the mannose receptor in infection of macrophages by influenza virus. AB - Influenza viruses A/PR/8/34 (PR8; H1N1), A/Aichi/68 X-31 (HKx31; H3N2), and A/Beijing/89 X-109 (BJx109; H3N2) show marked differences in their ability to infect murine macrophages, including resident alveolar and peritoneal macrophages as well as the macrophage-derived cell line J774. The hierarchy in infectivity of the viruses (PR8 < HKx31 < BJx109) resembles that of their reactivity with mannose-binding lectins of the collectin family. Since the macrophage mannose receptor recognizes the same spectrum of monosaccharides as the collectins do, we investigated the possible involvement of this receptor in infection of macrophages by influenza virus. In competitive binding studies, the binding of (125)I-labeled mannosylated bovine serum albumin to macrophages was inhibited by the purified hemagglutinin and neuraminidase (HANA) glycoproteins of influenza virus but not by HANA that had been treated with periodate to oxidize its oligosaccharide side chains. The inhibitory activity of HANA from the three strains of virus differed markedly and correlated with the infectivity of each virus for macrophages. Infection of macrophages, but not MDCK cells, by influenza virus was inhibited by yeast mannan. A variant line of J774 cells, J774E, which expresses elevated levels of the mannose receptor, was more readily infected than J774, and the sensitivity of J774E cells to infection was greatly reduced by culture in the presence of D-mannose, which down-modulated mannose receptor expression. Together, the data implicate the mannose receptor as a major endocytic receptor in the infectious entry of influenza virus, and perhaps other enveloped viruses, into murine macrophages. PMID- 10799596 TI - Influenza A viruses lacking sialidase activity can undergo multiple cycles of replication in cell culture, eggs, or mice. AB - Influenza A viruses possess both hemagglutinin (HA), which is responsible for binding to the terminal sialic acid of sialyloligosaccharides on the cell surface, and neuraminidase (NA), which contains sialidase activity that removes sialic acid from sialyloligosaccharides. Interplay between HA receptor-binding and NA receptor-destroying sialidase activity appears to be important for replication of the virus. Previous studies by others have shown that influenza A viruses lacking sialidase activity can undergo multiple cycles of replication if sialidase activity is provided exogenously. To investigate the sialidase requirement of influenza viruses further, we generated a series of sialidase deficient mutants. Although their growth was less efficient than that of the parental NA-dependent virus, these viruses underwent multiple cycles of replication in cell culture, eggs, and mice. To understand the molecular basis of this viral growth adaptation in the absence of sialidase activity, we investigated changes in the HA receptor-binding affinity of the sialidase deficient mutants. The results show that mutations around the HA receptor-binding pocket reduce the virus's affinity for cellular receptors, compensating for the loss of sialidase. Thus, sialidase activity is not absolutely required in the influenza A virus life cycle but appears to be necessary for efficient virus replication. PMID- 10799595 TI - Yin yang 1 negatively regulates the differentiation-specific E1 promoter of human papillomavirus type 6. AB - Human papillomavirus type 6 (HPV-6) is a low-risk HPV whose replication cycle, like that of all HPVs, is differentiation dependent. We have previously shown that CCAAT displacement protein (CDP) binds the differentiation-induced HPV-6 E1 promoter and negatively regulates its activity in undifferentiated cells (W. Ai, E. Toussaint, and A. Roman, J. Virol. 73:4220-4229, 1999). Using electrophoretic mobility shift assays (EMSAs), we now report that Yin Yang 1 (YY1), a multifunctional protein that can act as a transcriptional activator or repressor and that can also inhibit HPV replication in vitro, binds the HPV-6 E1 promoter. EMSAs, using subfragments of the promoter as competitors, showed that the YY1 binding site is located at the 5' end of the E1 promoter. When a putative YY1 site was mutated, the ability of YY1 to bind was greatly decreased. The activity of the mutated E1 promoter, monitored with the reporter gene luciferase, was threefold greater than that of the wild-type promoter, suggesting that YY1 negatively regulates HPV-6 E1 promoter activity. Nuclear extracts from differentiated keratinocytes showed decreased binding of YY1 to the wild-type promoter. Consistent with this, in differentiated keratinocytes, the activity of the transfected luciferase gene transcribed from the mutated promoter was comparable to that of the wild-type promoter; both promoters were up-regulated in differentiated keratinocytes compared to undifferentiated cells. These data suggest that YY1 functions in undifferentiated keratinocytes but not in differentiated keratinocytes. Both the wild-type and mutated promoters could be negatively regulated by overexpression of a plasmid encoding CDP. Thus, both YY1 and CDP appear to be negative regulators of the differentiation-induced HPV-6 E1 promoter and thereby the HPV life cycle. In contrast, only binding of CDP was detected using the E1 promoter of the high-risk HPV-31. PMID- 10799597 TI - The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis. AB - Equine arteritis virus (EAV), the prototype Arterivirus, is a positive-stranded RNA virus that expresses its replicase in the form of two large polyproteins of 1,727 and 3,175 amino acids. The functional replicase subunits (nonstructural proteins), which drive EAV genome replication and subgenomic mRNA transcription, are generated by extensive proteolytic processing. Subgenomic mRNA transcription involves an unusual discontinuous step and generates the mRNAs for structural protein expression. Previously, the phenotype of mutant EAV030F, which carries a single replicase point mutation (Ser-2429-->Pro), had implicated the nsp10 replicase subunit (51 kDa) in viral RNA synthesis, and in particular in subgenomic mRNA transcription. nsp10 contains an N-terminal (putative) metal binding domain (MBD), located just upstream of the Ser-2429-->Pro mutation, and a helicase activity in its C-terminal part. We have now analyzed the N-terminal domain of nsp10 in considerable detail. A total of 38 mutants, most of them carrying specific single point mutations, were tested in the context of an EAV infectious cDNA clone. Variable effects on viral genome replication and subgenomic mRNA transcription were observed. In general, our results indicated that the MBD region, and in particular a set of 13 conserved Cys and His residues that are assumed to be involved in zinc binding, is essential for viral RNA synthesis. On the basis of these data and comparative sequence analyses, we postulate that the MBD may employ a rather unusual mode of zinc binding that could result in the association of up to four zinc cations with this domain. The region containing residue Ser-2429 may play the role of "hinge spacer," which connects the MBD to the rest of nsp10. Several mutations in this region specifically affected subgenomic mRNA synthesis. Furthermore, one of the MBD mutants was replication and transcription competent but did not produce infectious progeny virus. This suggests that nsp10 is involved in an as yet unidentified step of virion biogenesis. PMID- 10799598 TI - Topoisomerase I associates specifically with simian virus 40 large-T-antigen double hexamer-origin complexes. AB - Topoisomerase I (topo I) is required for releasing torsional stress during simian virus 40 (SV40) DNA replication. Recently, it has been demonstrated that topo I participates in initiation of replication as well as in elongation. Although T antigen and topo I can bind to one another in vitro, there is no direct evidence that topo I is a component of the replication initiation complex. We demonstrate in this report that topo I associates with T-antigen double hexamers bound to SV40 origin DNA (T(DH)) but not to single hexamers. This association has the same nucleotide and DNA requirements as those for the formation of double hexamers on DNA. Interestingly, topo I prefers to bind to fully formed T(DH) complexes over other oligomerized forms of T antigen associated with the origin. High ratios of topo I to origin DNA destabilize T(DH). The partial unwinding of a small-circular DNA substrate is dependent on the presence of both T antigen and topo I but is inhibited at high topo I concentrations. Competition experiments with a topo I binding fragment of T antigen indicate that an interaction between T antigen and topo I occurs during the unwinding reaction. We propose that topo I is recruited to the initiation complex after the assembly of T(DH) and before unwinding to facilitate DNA replication. PMID- 10799599 TI - Nonstructural protein 5A of hepatitis C virus inhibits the function of karyopherin beta3. AB - It has been suggested that nonstructural protein 5A (NS5A) of hepatitis C virus (HCV) plays a role in the incapacitation of interferon by inactivation of RNA dependent protein kinase PKR. In order to further investigate the role of NS5A, we tried to identify cellular proteins interacting with NS5A by using the yeast two-hybrid system. The karyopherin beta3 gene was isolated from a human liver cell library as a protein interacting with NS5A. The protein-protein interaction between NS5A and karyopherin beta3 was confirmed by in vitro binding assay and an in vivo coimmunoprecipitation method. The effect of NS5A on the karyopherin beta3 activity was investigated using a yeast cell line containing mutations in both PSE1 and KAP123, genes that are homologous to the human karyopherin beta3 gene. Human karyopherin beta3 complemented the loss of the PSE1 and KAP123 functions, supporting growth of the double mutant cells. However, expression of NS5A hampered the growth of the double mutant cells supplemented with human karyopherin beta3. On the other hand, expression of NS5A by itself had no effect on the growth of the double mutant expressing wild-type yeast PSE1. This indicates that NS5A may inhibit karyopherin beta3 function via protein-protein interaction. The role of NS5A in HCV replication is discussed. PMID- 10799600 TI - Chimeric retroviral helper virus and picornavirus IRES sequence to eliminate DNA methylation for improved retroviral packaging cells. AB - Most retroviral packaging cell lines were established by a helper virus plasmid cotransfected with a separate plasmid encoding a selection marker. Since this selection marker coexisted in trans with the helper virus sequence, helper virus gene expression could be inactivated by host DNA methylation despite selection for the cotransfected selection marker. We have reported that DNA methylation could occur in the long terminal repeat (LTR) region of helper virus in vector producer cells (VPC) in up to 2% of the population per day (W. B. Young, G. L. Lindberg, and C. J. Link, Jr., J. Virol. 74:3177-3187, 2000). To overcome host cell DNA methylation that suppresses viral gene expression, we constructed a chimeric retroviral helper virus, pAM3-IRES-Zeo, that contains Moloney murine leukemia virus as a helper virus and a picornavirus internal ribosome entry site (IRES) sequence followed by a Zeocin selection marker at the 3' end of the env sequence. This pAM3-IRES-Zeo permitted selection for intact and functional helper virus in transfected cells without subcloning. By selection with Zeocin, a mixed population of pAM3-IRES-Zeo-transfected NIH3T3 cells (AMIZ cells) was maintained with little or no DNA methylation of the helper virus 5' LTR. The high level of pAM3-IRES-Zeo gene expression resulted in no detectable vector superinfection and in high vector titers (2 x 10(6) to 1.5 x 10(7) CFU/ml) after introduction of a retroviral vector. When Zeocin selection was withdrawn from AMIZ cells, methylation of the 5' LTR increased from 17 to 36% of the population during 67 days of continuous culture and the cells became susceptible to superinfection. During this period, gene expression of pAM3-IRES-Zeo decreased and vector titer production was reduced to 2 x 10(4) CFU/ml. These data demonstrate an important role of DNA methylation in the genetic instability of VPC. The chimeric helper virus allows the establishment of a mixed population of packaging cells capable of high-level and sustained vector production without cloning procedures. PMID- 10799601 TI - Protective immunity to rotavirus shedding in the absence of interleukin-6: Th1 cells and immunoglobulin A develop normally. AB - We investigated whether interleukin-6 (IL-6) was required for the development of immunoglobulin A (IgA)- and T-helper 1 (Th1)-associated protective immune responses to rotavirus by using adult IL-6-deficient mice [BALB/c and (C57BL/6 x O1a)F(2) backgrounds]. Naive IL-6(-) mice had normal frequencies of IgA plasma cells in the gastrointestinal tract. Consistent with this, total levels of IgA in fecal extracts, saliva, and sera were unaltered. In specific response to oral infection with rhesus rotavirus, IL-6(-) and IL-6(+) mice exhibited efficient Th1 type gamma interferon responses in Peyer's patches with high levels of serum IgG2a and intestinal IgA. Although there was an increase in Th2-type IL-4 in CD4(+) T cells from IL-6(-) mice following restimulation with rotavirus antigen in the presence of irradiated antigen-presenting cells, unfractionated Peyer's patch cells failed to produce a significant increase in IL-4. Moreover, virus specific IgG1 in serum was not significantly increased in IL-6(-) mice in comparison with IL-6(+) mice. Following oral inoculation with murine rotavirus, IL-6(-) and IL-6(+) mice mediated clearance of rotavirus and mounted a strong IgA response. When IL-6(-) and IL-6(+) mice [(C57BL/6 x O1a)F(2) background] were orally inoculated with rhesus rotavirus and later challenged with murine rotavirus, all of the mice maintained high levels of IgA in feces and were protected against reinfection. Thus, IL-6 failed to provide unique functions in the development of IgA-secreting B cells and in the establishment of Th1 associated protective immunity against rotavirus infection in adult mice. PMID- 10799602 TI - Low dynamic state of viral competition in a chronic avian hepadnavirus infection. AB - The dynamic state of infection of 11 ducks with the duck hepatitis B virus was investigated. Chronic infections were established in newly hatched ducklings by inoculation with a mixture of wild-type virus and a mutant virus with a partial replication defect. As expected, the wild-type virus was rapidly enriched in the virus population during the spread of infection. Enrichment thereafter was correlated with normal growth of the liver, with the average mutant-to-wild-type ratio stabilizing for at least 2 months beyond the time at which the liver mass stabilized. Using experimentally determined growth rates for the mutant and wild type viruses, we estimated that after the spread of infection, competition between the two virus strains was limited by the amount of replication required to infect new hepatocytes in the growing livers. The results suggest that, in a chronically infected liver, the selection of variants with a replication rate advantage is inefficient and that the emergence of such variants would depend on induced liver cell turnover, such as that occurring during chronic hepatitis. PMID- 10799604 TI - The Autographa californica nuclear polyhedrosis virus p143 gene encodes a DNA helicase. AB - The P143 protein of Autographa californica nuclear polyhedrosis virus is essential for replication of viral DNA. To determine the function of P143, the protein was purified to near homogeneity from recombinant baculovirus-infected cells that overexpress P143. ATPase activity copurified with P143 protein during purification and also during gel filtration at a high salt concentration. The ATPase activity did not require the presence of single-stranded DNA, but was stimulated fourfold by the addition of single-stranded DNA. The ATPase activity of P143 had a K(m) of 60 microM and a turnover of 4.5 molecules of ATP hydrolyzed/s/molecule of enzyme, indicating moderate affinity for ATP and high catalytic efficiency. P143 unwound a 40-nucleotide primer in an ATP-dependent manner, indicating that the enzyme possesses in vitro DNA helicase activity. Based on this result, it seems likely that P143 functions as a helicase in viral DNA replication. PMID- 10799603 TI - Expression of hepatitis B virus X protein does not alter the accumulation of spontaneous mutations in transgenic mice. AB - Chronic infection with hepatitis B virus (HBV) is one of the major etiological factors in the development of human hepatocellular carcinoma. Transgenic mice that express the HBV X protein (HBx) have previously been shown to be more sensitive to the effects of hepatocarcinogens, although the mechanism for this cofactor role remains unknown. The ability of HBx to inhibit DNA repair in transiently transfected cell lines suggests one possible pathway. In the present study, primary hepatocytes isolated from transgenic mice that possess the HBV X gene under the control of the human alpha-1-antitrypsin regulatory region (ATX mice) were found to be deficient in their ability to conduct unscheduled DNA synthesis in response to UV-induced DNA damage. In order to measure the impact of HBx expression on DNA repair in vivo, double-transgenic mice that express HBx and possess a bacteriophage lambda transgene were sacrificed at 30, 90, and 240 days of age. Mutation frequency was determined for high-molecular-weight liver DNA of ATX and control mice by functional analysis of the lambda transgene. Expression of HBx did not significantly increase the accumulation of spontaneous mutations. These results are consistent with previous studies of HBx transgenic mice in which no effect of HBx on liver histology was apparent. This new animal model provides a powerful system in which to investigate the in vivo cooperation between HBx expression and environmental carcinogens. PMID- 10799605 TI - J domain-independent regulation of the Rb family by polyomavirus large T antigen. AB - The ability of polyomavirus large T antigen (LT) to promote cell cycling, to immortalize primary cells, and to block differentiation has been linked to its effects on tumor suppressors of the retinoblastoma susceptibility (Rb) gene family. Our previous studies have shown that LT requires an intact N-terminal DnaJ domain, in addition to an Rb binding site, for activation of simple E2F containing promoters and stimulation of cell cycle progression. Here we show that some LT effects dependent on interaction with the Rb family are largely DnaJ independent. In differentiating C2C12 myoblasts, overexpression of LT caused apoptosis. Although this activity of LT completely depended on Rb binding, LTs with mutations in the J domain remained able to kill. Comparisons of Rb(-) and J( ) LTs revealed additional differences. Wild-type but not Rb(-) LT activated the cyclin A promoter under serum starvation conditions. Genetic analysis of the promoter linked the Rb requirement to an E2F site in the promoter. LTs with mutations in the J domain were still able to activate the promoter. Finally, J mutant LTs caused changes in phosphorylation of both pRb and p130. In the case of p130, Thr-986 was shown to be a site that is regulated by J mutant LT. Taken together, these observations reveal that LT regulation of Rb function can be separated into both DnaJ-dependent and DnaJ-independent pathways. PMID- 10799607 TI - Downregulation of major histocompatibility complex class I molecules by Kaposi's sarcoma-associated herpesvirus K3 and K5 proteins. AB - The T-cell-mediated immune response plays a central role in the defense against intracellular pathogens. To avoid this immune response, viruses have evolved elaborate mechanisms that target and modulate many different aspects of the host's immune system. A target common to many of these viruses is the major histocompatibility complex (MHC) class I molecules. Kaposi's sarcoma-associated herpesvirus (KSHV) encodes K3 and K5 zinc finger membrane proteins which remove MHC class I molecules from the cell surface. K3 and K5 exhibit 40% amino acid identity to each other and localize primarily near the plasma membrane. While K3 and K5 dramatically downregulated class I molecules, they displayed different specificities in downregulation of HLA allotypes. K5 significantly downregulated HLA-A and -B and downregulated HLA-C only weakly, but not HLA-E, whereas K3 downregulated all four HLA allotypes. This selective downregulation of HLA allotypes by K5 was partly due to differences in amino acid sequences in their transmembrane regions. Biochemical analyses demonstrated that while K3 and K5 did not affect expression and intracellular transport of class I molecules, their expression induced rapid endocytosis of the molecules. These results demonstrate that KSHV has evolved a novel immune evasion mechanism by harboring similar but distinct genes, K3 and K5, which target MHC class I molecules in different ways. PMID- 10799606 TI - HLA-B57-restricted cytotoxic T-lymphocyte activity in a single infected subject toward two optimal epitopes, one of which is entirely contained within the other. AB - Viral peptides are recognized by cytotoxic T lymphocytes (CTL) as a complex with major histocompatibility complex (MHC) class I molecules, but the extent to which a single HLA allele can accommodate epitope peptides of different length and sequence is not well characterized. Here we report the identification of clonal CTL responses from the same donor that independently recognize one of two HLA-B57 restricted epitopes, KAFSPEVIPMF (KF11; p24(Gag) residues 30 to 40) and KAFSPEVI (KF8; p24(Gag) residues 30 to 37). Although lysis studies indicated that the KF11 peptide stabilized the HLA-B57-peptide complex more efficiently than the KI8 peptide, strong clonal responses were directed at each epitope. In samples from a second donor, the same phenomenon was observed, in which distinct CTL clones recognized peptide epitopes presented by the same HLA class I allele (in this case, HLA-A3) which were entirely overlapping. These data are relevant to the accurate characterization of CTL responses, which is fundamental to a detailed understanding of MHC class I-restricted immunity. In addition, these studies demonstrate marked differences in the length of peptides presented by HLA-B57, an allele which is associated with nonprogressive human immunodeficiency virus infection. PMID- 10799608 TI - Mutation of a conserved residue (D123) required for oligomerization of human immunodeficiency virus type 1 Nef protein abolishes interaction with human thioesterase and results in impairment of Nef biological functions. AB - Nef is a myristoylated protein of 27 to 35 kDa that is conserved in primate lentiviruses. In vivo, Nef is required for high viral load and full pathological effects. In vitro, Nef has at least four activities: induction of CD4 and major histocompatibility complex (MHC) class I downregulation, enhancement of viral infectivity, and alteration of T-cell activation pathways. We previously reported that the Nef protein from human immunodeficiency virus type 1 interacts with a novel human thioesterase (hTE). In the present study, by mutational analysis, we identified a region of the Nef core, extending from the residues D108 to W124, that is involved both in Nef-hTE interaction and in Nef-induced CD4 downregulation. This region of Nef is located on the oligomer interface and is in close proximity to the putative CD4 binding site. One of the mutants carrying a mutation in this region, targeted to the conserved residue D123, was also found to be defective in two other functions of Nef, MHC class I downmodulation and enhancement of viral infectivity. Furthermore, mutation of this residue affected the ability of Nef to form dimers, suggesting that the oligomerization of Nef may be critical for its multiple functions. PMID- 10799609 TI - Incorporation of tumor vasculature targeting motifs into moloney murine leukemia virus env escort proteins enhances retrovirus binding and transduction of human endothelial cells. AB - Adhesion receptors expressed on the surfaces of tumor-activated endothelial cells provide an advantageous locus for targeting gene therapy vectors to angiogenic tissues and/or tumor vasculature. In this study, we engineered a series of Asn Gly-Arg (NGR)-containing congeners of the presumptive cell binding motif contained within the ninth type III repeat of fibronectin and displayed these tumor vasculature targeting motifs (TVTMs) within the context of Moloney murine leukemia envelope "escort" proteins. Comparative studies of envelope incorporation into viral particles and evaluation of the cell binding properties of the targeted vectors revealed critical structural features, thus identifying a subset of optimal TVTMs. Utilizing a modified ELISA to evaluate viral binding to target cells, we observed a significant down-regulation of TVTM-virion binding to human endothelial cells following sustained (48-h) exposure to VEGF. Normalized for equivalent titers (10(6) CFU/ml), as assayed on NIH 3T3 cells, vectors displaying TVTM escort proteins significantly enhanced the transduction efficiency from 12.2 to 37.4% in human KSY-1 endothelial cell cultures (P < 0.001) and from 0.4 to 4.1% in human umbilical vein endothelial cell (HUVEC) cultures (P < 0.001). In summary, these studies utilized an engineering approach to identify a subset of TVTMs that are stably incorporated as envelope "escort" proteins into retroviral vectors and that, by functioning to improve the binding efficiency and transduction of both HUVEC and KSY1 endothelial cells, may have therapeutic potential for targeting gene delivery to the tumor-associated vasculature. PMID- 10799610 TI - Hepatitis delta virus replication generates complexes of large hepatitis delta antigen and antigenomic RNA that affiliate with and alter nuclear domain 10. AB - Hepatitis delta virus (HDV), a single-stranded RNA virus, bears a single coding region whose product, the hepatitis delta antigen (HDAg), is expressed in two isoforms, small (S-HDAg) and large (L-HDAg). S-HDAg is required for replication of HDV, while L-HDAg inhibits viral replication and is required for the envelopment of the HDV genomic RNA by hepatitis B virus proteins. Here we have examined the spatial distribution of HDV RNA and proteins in infected nuclei, with particular reference to specific nuclear domains. We found that L-HDAg was aggregated in specific nuclear domains and that over half of these domains were localized beside nuclear domain 10 (ND10). At later times, ND10-associated proteins like PML were found in larger HDAg complexes that had developed into apparently hollow spheres. In these larger complexes, PML was found chiefly in the rims of the spheres, while the known ND10 components Sp100, Daxx, and NDP55 were found in the centers of the spheres. Thus, ND10 proteins that normally are closely linked separate within HDAg-associated complexes. Viral RNA of antigenomic polarity, whether expressed from genomic RNA or directly from introduced plasmids, colocalizes with L-HDAg and the transcriptional repressor PML. In contrast, HDV genomic RNA was distributed more uniformly throughout the nucleus. These results suggest that different host protein complexes may assemble on viral RNA strands of different polarities, and they also suggest that this RNA virus, like DNA viruses, can alter the distribution of ND10-associated proteins. The fact that viral components specifically linked to repression of replication can associate with one of the ND10-associated proteins (PML) raises the possibility that this host protein may play a role in the regulation of HDV RNA synthesis. PMID- 10799611 TI - Persistence and reactivation of bovine herpesvirus 1 in the tonsils of latently infected calves. AB - Bovine herpesvirus 1 (BHV-1), like other members of the Alphaherpesvirinae subfamily, establishes latent infection in sensory neurons. Reactivation from latency can occur after natural or corticosteroid-induced stress culminating in recurrent disease and/or virus transmission to uninfected animals. Our previous results concluded that CD4(+) T cells in the tonsil and other adjacent lymph nodes are infected and undergo apoptosis during acute infection (M. T. Winkler, A. Doster, and C. Jones, J. Virol. 73:8657-8668, 1999). To test whether BHV-1 persisted in lymphoreticular tissue, we analyzed tonsils of latently infected calves for the presence of viral DNA and gene expression. BHV-1 DNA was consistently detected in the tonsils of latently infected calves. Detection of the latency-related transcript (LRT) in tonsils of latently infected calves required nested reverse transcription-PCR (RT-PCR) suggesting that only a few cells contained viral DNA or that LRT is not an abundant transcript. bICP0 (immediate-early and early transcripts), ribonucleotide reductase (early transcript), and glycoprotein C (late transcript) were not detected by RT-PCR in latently infected calves. When reactivation was initiated by dexamethasone, bICP0 and ribonucleotide reductase transcripts were detected. Following dexamethasone treatment, viral nucleic acid was detected simultaneously in trigeminal ganglionic neurons and lymphoid follicles of tonsil. LRT was detected at 6 and 24 h after dexamethasone treatment but not at 48 h. Dexamethasone-induced reactivation led to apoptosis that was localized to tonsillar lymphoid follicles. Taken together, these findings suggest that the tonsil is a site for persistence or latency from which virus can be reactivated by dexamethasone. We further hypothesize that the shedding of virus from the tonsil during reactivation plays a role in virus transmission. PMID- 10799612 TI - Human immunodeficiency virus type 1 coreceptor preferences determine target T cell depletion and cellular tropism in human lymphoid tissue. AB - The present study sought to determine how usage of coreceptors by human immunodeficiency virus type 1 dictates cell tropism and depletion of CD4(+) T cells in human lymphoid tissues cultured ex vivo. We found that coreceptor preferences control the marked, preferential depletion of coreceptor-expressing CD4(+) lymphocytes. In addition, there was a strong, but not absolute, preference shown by CXCR4-using strains for lymphocytes and by CCR5-using strains for macrophages. PMID- 10799613 TI - Activation of divergent neuronal cell death pathways in different target cell populations during neuroadapted sindbis virus infection of mice. AB - Infection of adult mice with neuroadapted Sindbis virus (NSV) results in a severe encephalomyelitis accompanied by prominent hindlimb paralysis. We find that the onset of paralysis parallels morphologic changes in motor neuron cell bodies in the lumbar spinal cord and in motor neuron axons in ventral nerve roots, many of which are eventually lost over time. However, unlike NSV-induced neuronal cell death found in the brain of infected animals, the loss of motor neurons does not appear to be apoptotic, as judged by morphologic and biochemical criteria. This may be explained in part by the lack of detectable caspase-3 expression in these cells. PMID- 10799614 TI - Convergent evolution of reverse transcriptase (RT) genes of human immunodeficiency virus type 1 subtypes E and B following nucleoside analogue RT inhibitor therapies. AB - Changes in the drug susceptibility, gene lineage, and deduced amino acid sequences of the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) subtype E following 3'-azido-3'-deoxythymidine (AZT) monotherapy or AZT 2', 3'-dideoxyinosine combination therapy were examined with sequential virus isolates from a single family. The changes were compared to those reported for HIV-1 subtype B, revealing striking similarities in selected phenotype and amino acids independent of differences in the RT backbone sequences that constantly distinguish the two subtypes. Particularly, identical amino acid substitutions were present simultaneously at four different positions (D67N, K70R, T215F, and K219Q) for high-level AZT resistance. These data suggest that HIV-1 subtypes E and B evolve convergently at the phenotypic and amino acid levels when the nucleoside analogue RT inhibitors act as selective forces. PMID- 10799615 TI - Major histocompatibility complex class I gene controls the generation of gamma interferon-producing CD4(+) and CD8(+) T cells important for recovery from friend retrovirus-induced leukemia. AB - Recovery from leukemia induced by Friend virus complex (FV) requires strong CD4(+) helper, CD8(+) cytotoxic T-lymphocyte, and B-cell responses. The development of these immune responses is dependent on the major histocompatibility complex (MHC) (H-2) genotype of the mouse. In H-2(b/b) mice, which spontaneously recover from FV-induced erythroleukemia, neutralization of gamma interferon (IFN-gamma) in vivo inhibited recovery, which indicated that IFN gamma was a necessary component of the immune response to FV. Furthermore, in H 2(b/b) mice, high numbers of IFN-gamma-producing cells were detected after FV infection, whereas in H-2(a/b) mice, which have a low-recovery phenotype, only low numbers of IFN-gamma-producing cells were detected. Similarly, H-2(bm14/b) mice, which cannot recover from FV infection due to a point mutation in one allele of the H-2D(b) gene, also had low numbers of IFN-gamma-producing T cells. Surprisingly, this effect was observed for both CD8(+) and CD4(+) T cells. These findings reveal a novel influence of MHC class I genes on CD4(+) T-cell responses to viral infection. Furthermore, the influence of MHC class I genotype on the generation of both IFN-gamma-producing CD4(+) and CD8(+) T cells helps explain the major impact of the H-2D gene on recovery from FV disease. PMID- 10799616 TI - Structure-function studies of the self-assembly domain of the human immunodeficiency virus type 1 transmembrane protein gp41. AB - The coiled-coil region of the human immunodeficiency virus type 1 transmembrane protein (gp41) makes up the interior core of the six-helix bundle structure of the gp41 self-assembly domain. We extended our previous study of this domain (Y. Weng and C. D. Weiss, J. Virol. 72:9676-9682, 1998) by analyzing 23 additional mutants at positions that lie at the interface of the interior core and outer helices. We found nine new functional mutants. For most mutants, the activity could be explained by the ability of the modeled mutants to stabilize the six helix bundle structure. The present study provides insights into the envelope glycoprotein fusion mechanism and information for rational drug and vaccine design. PMID- 10799617 TI - Ubiquitination of the human immunodeficiency virus type 1 env glycoprotein. AB - Expression of the human immunodeficiency virus type 1 (HIV-1) Env glycoprotein is stringently regulated in infected cells. The majority of the glycoprotein does not reach the cell surface but rather is retained in the endoplasmic reticulum or a cis-Golgi compartment and subsequently degraded. We here report that Env of various HIV-1 isolates is ubiquitinated at the extracellular domain of gp41 and that Env expression could be increased by lactacystin, a specific proteasome inhibitor, suggesting that the ubiquitin/proteasome system is involved in control of expression and degradation. PMID- 10799618 TI - Molecular epidemiology of hepatitis B virus variants in nonhuman primates. AB - We characterized hepatitis B virus (HBV) isolates from sera of 21 hepatitis B virus surface antigen-positive apes, members of the families Pongidae and Hylobatidae (19 gibbon spp., 1 chimpanzee, and 1 gorilla). Sera originate from German, French, Thai, and Vietnamese primate-keeping institutions. To estimate the phylogenetic relationships, we sequenced two genomic regions, one located within the pre-S1/pre-S2 region and one including parts of the polymerase and the X protein open reading frames. By comparison with published human and ape HBV isolates, the sequences could be classified into six genomic groups. Four of these represented new genomic groups of gibbon HBV variants. The gorilla HBV isolate was distantly related to the chimpanzee isolate described previously. To confirm these findings, the complete HBV genome from representatives of each genomic group was sequenced. The HBV isolates from gibbons living in different regions of Thailand and Vietnam could be classified into four different phylogenetically distinct genomic groups. The same genomic groups were found in animals from European zoos. Therefore, the HBV infections of these apes might have been introduced into European primate-keeping facilities by direct import of already infected animals from different regions in Thailand. Taken together, our data suggest that HBV infections are indigenous in the different apes. One event involving transmission between human and nonhuman primates in the Old World of a common ancestor of human HBV genotypes A to E and the ape HBV variants might have occurred. PMID- 10799619 TI - Evaluation of a neonatal rat model for prediction of mumps virus neurovirulence in humans. AB - Neurovirulence of several mumps virus strains was assessed in a prototype rat neurovirulence test and compared to results obtained in the monkey neurovirulence test. The relative human neurovirulence of these strains was proportional to the severity of hydrocephalus in rats but not to lesion scores in the monkeys. PMID- 10799620 TI - A single amino acid change in the murine leukemia virus capsid gene responsible for the Fv1(nr) phenotype. AB - The nr allele at the mouse Fv1 restriction locus governs resistance to B-tropic and some N-tropic murine leukemia viruses (MLVs). Sequence analysis and site specific mutagenesis of N-tropic MLVs identified a single amino acid difference responsible for this restriction that is distinct from the site that governs N or B tropism. Viruses with other substitutions at this site were evaluated for altered replication patterns. PMID- 10799621 TI - Probing the structure of rotavirus NSP4: a short sequence at the extreme C terminus mediates binding to the inner capsid particle. AB - The rotavirus nonstructural glycoprotein NSP4 functions as the receptor for the inner capsid particle (ICP) which buds into the lumen of the endoplasmic reticulum during virus maturation. The structure of the cytoplasmic domain of NSP4 from rotavirus strain SA11 has been investigated by using limited proteolysis and mass spectrometry. Digestion with trypsin and V8 protease reveals a C-terminal protease-sensitive region that is 28 amino acids long. The minimal sequence requirements for receptor function have been defined by constructing fusions with glutathione S-transferase and assessing their ability to bind ICPs. These experiments demonstrate that 17 to 20 amino acids from the extreme C terminus are necessary and sufficient for ICP binding and that this binding is cooperative. These observations are consistent with a model for the structure of the NSP4 cytoplasmic region in which four flexible regions of 28 amino acids are presented by a protease-resistant coiled-coil tetramerization domain, with only the last approximately 20 amino acids of each peptide interacting with the surface binding sites on the ICP. PMID- 10799622 TI - What is the yield of screening echocardiography in pediatric syncope? AB - OBJECTIVE: To determine the yield of screening echocardiography in the evaluation of pediatric syncope. DESIGN: All patients diagnosed with syncope from January 1993 to January 1999 were identified and their records were reviewed for age, weight, sex, year of presentation, personal and family history, physical examination, and cardiac diagnostic testing. Cardiac defects were identified by reviewing echocardiograms and reports. RESULTS: The 480 patients (268 females) ranged in age from 1.5 to 18.0 years old and ranged in weight from 10.3 to 113.6 kg. Final diagnoses included noncardiac causes in 458, long QT syndrome in 14, arrhythmias in 6, and cardiomyopathy in 2. An abnormal history, physical examination, or electrocardiogram identified 21 of the 22 patients with a cardiac cause of syncope. Of the 322 (67%) echocardiograms performed, abnormalities were detected in 37. These abnormalities included 26 minor valve anomalies, 7 hemodynamically insignificant shunt lesions, 2 mildly decreased left ventricular shortening fractions, and 2 cardiomyopathies. Only the 2 cardiomyopathies were considered to be potential causes of syncope, and in both cases, the electrocardiogram was markedly abnormal. A similar percentage of echocardiograms were ordered during the first and last 3 years of the study (61% vs 71%). CONCLUSION: History, physical examination, and electrocardiography provide a screening protocol that allows the identification of a cardiac cause of syncope in the overwhelming majority of pediatric patients. In the absence of a positive screen result, the echocardiogram does not contribute to the evaluation of syncope in children. We speculate that primary care providers and pediatric cardiologists continue to use echocardiography because of the paucity of data regarding its value in pediatric syncope. However, this study shows little benefit of screening echocardiography and should discourage its routine use. PMID- 10799623 TI - Nonresponders: prolonged fever among infants with urinary tract infections. AB - BACKGROUND: The majority of young children with fever and urinary tract infections (UTIs) have evidence of pyelonephritis based on renal scans. Resolution of fever during treatment is 1 clinical marker of adequate treatment. Theoretically, prolonged fever may be a clue to complications, such as urinary obstruction or renal abscess. OBJECTIVE: Describe the pattern of fever in febrile children undergoing treatment of a UTI. Compare the clinical characteristics of those patients with prolonged fever to those who respond faster to therapy. SETTING: An urban pediatric hospital. DESIGN: Medical record review. METHODS: All children /=38 degrees C and those who met standard culture criteria were studied. Temperatures are not recorded hourly on the inpatient unit; therefore, they were assigned to blocks of time. Nonresponders were defined as those above the 90th percentile for the time to defervesce. Nonresponders were then compared with the balance of the study patients, termed responders. RESULTS: Of 288 patients studied, the median age was 5.6 months (interquartile range: 1.3-7.9 months old). Median admission temperature was 39.3 degrees C (interquartile range: 38.5 degrees C-40.1 degrees C). Median time to defervesce ranged in the time block 13 to 16 hours. Sixty eight percent were afebrile by 24 hours and 89% by 48 hours. Thirty-one patients had fever >48 hours (nonresponders). Nonresponders were older than responders (9.4 vs 4.1 months old) but had similar initial temperatures (39.8 vs 39.2 degrees C), white blood cell counts (18.4 vs 17.1 x 1000/mm(3)), and band counts (1.4 vs 1.2 x 1000/mm(3)). Nonresponders had similar urinalyses with regard to leukocyte esterase positive (23/29 vs 211/246), nitrite-positive (8/28 vs 88/221], and the number of patients with "too numerous to count" white blood cell counts per high power field (12/28 vs 77/220). Nonresponders were as likely as responders to have bacteremia (3/31 vs 21/256), hydronephrosis by renal ultrasound (1/31 vs 12/232), and significant vesicoureteral reflux (more than or equal to grade 3; 5/26 vs 30/219). Eschericia coli was the pathogen in cultures of 28 of 31 (nonresponders) and 225 of 257 (responders) cultures. The number of cultures with >/=100 colony-forming units/mL was similar (25/31 nonresponders vs 206/257 responders). Repeat urine cultures were performed in 93% of patients during the admission; all culture results were negative. No renal abscesses or pyo-hydronephrosis was diagnosed. CONCLUSIONS: Eighty-nine percent of young children with febrile UTIs were afebrile within 48 hours of initiating parenteral antibiotics. The patients who took longer than 48 hours to defervesce were clinically similar to those whose fevers responded faster to therapy. If antibiotic sensitivities are known, additional diagnostic studies or prolonged hospitalizations may not be justified solely based on persistent fever beyond 48 hours of therapy. PMID- 10799624 TI - Invasive group A streptococcal disease in children and association with varicella zoster virus infection. Ontario Group A Streptococcal Study Group. AB - OBJECTIVES: To describe the incidence and clinical features of invasive group A streptococcal (GAS) disease in children in Ontario and determine the risk of invasive GAS infection following chickenpox. METHODS: During 1992-1996, we conducted prospective, active, population-based surveillance for pediatric invasive GAS disease in Ontario, Canada (population: 11 million; 2.5 million children) and reviewed clinical and laboratory records. RESULTS: There were 1.9 cases of invasive GAS disease per 100,000 children per year. Streptococcal toxic shock syndrome (STSS) occurred in 7% of cases and necrotizing fasciitis (NF) in 4% for incidences of.08 and.13 per 100,000 per year, respectively. Case-fatality rates were 56% for STSS, 10% for NF, and 4% overall. The presence of chronic underlying illness other than asthma was associated with death (relative risk [RR]: 11; 95% confidence interval [CI]: 2.4-45). Fifteen percent of children identified had preceding chickenpox infection, which significantly increased the risk for acquisition of invasive GAS disease (RR: 58; 95% CI: 40-85). Children with invasive GAS and recent chickenpox were more likely to have NF (RR: 6.3; 95% CI: 1.8-22.3). CONCLUSIONS: Childhood invasive GAS disease occurs at an incidence similar to the adult population but has a lower rate of STSS and case-fatality. Chickenpox dramatically increases the risk for acquiring invasive GAS disease, and universal chickenpox vaccination could potentially prevent up to 15% of all pediatric invasive GAS disease. PMID- 10799625 TI - A prospective search for urinary tract abnormalities in infants with isolated preauricular tags. AB - OBJECTIVE: To determine whether isolated preauricular tags are associated with urinary tract abnormalities. METHODOLOGY: Seventy consecutive infants with isolated preauricular tags were examined by ultrasonography for urinary tract abnormalities on day 3 or 4 of life between January 1993 and August 1999, after parental consent and ethics approval. Karyotype analysis was conducted in all infants with urinary tract abnormalities. The study group was compared with a control group of 69 infants without preauricular tags hospitalized during the same period. The control group consisted of infants who underwent urinary tract ultrasonography as part of an investigation for persistent regurgitation and/or vomiting associated with cyanotic spells. RESULTS: Urinary tract abnormalities were detected in 6 infants with isolated preauricular tags (6/70; 8.6%). Types of anomalies were as follows: hydronephrosis in 5 cases and horseshoe kidney in 1 case. The causes of hydronephrosis were ureteropelvic junction obstruction in 3 cases and vescicoureteric reflux in 2 cases. None of the infants in the control group had such abnormalities. All infants with urinary tract abnormalities had normal chromosomes. No statistically significant differences existed between groups concerning birth weight, gestational age, intrauterine growth, and male-to female ratio. CONCLUSIONS: This study suggests a significant prevalence of urinary tract abnormalities in infants with preauricular tags. We recommend, therefore, that urinary tract ultrasonography be conducted in the routine assessment of infants with isolated preauricular tags. PMID- 10799626 TI - A family with extrinsic allergic alveolitis caused by wild city pigeons: A case report. AB - We describe a family in which the mother died of unresolved lung disease and whose 5 children, some of whom had previous signs of asthma, were subsequently affected by extrinsic allergic alveolitis caused by contact with wild city pigeon antigens. The children received systemic corticosteroids for 1 month and inhaled steroids for 24 months, while antigen exposure was reduced as much as feasible. This was followed by a quick clinical recovery and a slow normalization of chest radiographs and pulmonary function indices, especially of diffusion capacity, during a follow-up of 24 months. Because pigeon-breeder's lung caused by free roaming city pigeons has not been previously described, it remains unclear whether this family developed the disease because of high antigen exposure or because of increased susceptibility. None of the supposedly high-risk human leukocyte antigen types were found in the children. Whether human leukocyte antigen B7 in 1 child played a role in the course of the illness remains speculative. It is unknown to what extent pigeon-breeder's lung caused by nondomestic birds remains undetected and misdiagnosed as difficult or steroid resistant asthma. The question remains whether free-roaming city pigeons are indeed a public health risk. We suggest that atypical outdoor antigens be considered in all patients with nonresolving chest disease or therapy-resistant asthma. PMID- 10799627 TI - Predicting heart growth during puberty: The Muscatine Study. AB - OBJECTIVES: During childhood, heart growth is closely associated with somatic growth including increases in body weight, fat-free body mass (FFM), and height. However, with age, greater variability in heart size in relationship to body size is observed, presumably attributable to the increased effect of cardiac workload. At this time, little is known as to what functional attributes (eg, aerobic fitness) contribute to cardiac workload and the relative contribution of these attributes to heart growth during childhood and adolescence. In this article, we report cross-sectional and longitudinal relationships among aerobic fitness, body size, blood pressure (BP), and left ventricular mass (LVM) through puberty including the predictors of heart growth during puberty and the tracking of LVM from pre-puberty to late and post-puberty. Describing the predictors of heart size and heart growth and establishing the likelihood that a large heart, relative to peers, may (or may not) remain a large heart should aid pediatricians in discerning between normal developmental increases in LVM and increases in LVM suggestive of excessive heart growth (left ventricular hypertrophy). METHODOLOGY: Using a repeated-measures design, we assessed aerobic fitness, FFM, fatness, weight, height, sexual maturation, resting BP, peak exercise BP, and LVM in 125 healthy children (mean baseline age: 10.5 years) for a period of 5 years. All subjects were either in prepuberty or early puberty at the beginning of the study. At follow-up, 110 subjects attempted all research procedures (87% of the initial cohort). Using anthropometry and bioelectrical impedance, we measured FFM, fatness, weight, and height quarterly (once every 3 months) for a total of 20 examinations. Resting BP and LVM (2-dimensional echocardiography) were also assessed quarterly. Aerobic fitness, peak exercise BP, and sexual maturation (staging of secondary sex characteristics and, for boys, serum testosterone) were measured annually (5 examinations). The same field staff conducted all examinations. Statistical methods included Spearman rank correlation coefficients (r(s)) calculated to estimate how well the year 5 LVM was predicted by LVM at earlier years. We also categorized the LVM data into tertiles and reported the percentage who remained in the extreme tertiles in year 5, given they began in that tertile in year 1. Gender-specific stepwise multivariate analysis was used to evaluate predictors of follow-up LVM and predictors of changes in LVM. The latter model examined whether the variability in the changes in LVM, as quantified by subject-specific slopes, could be explained by changes in predictor variables, also quantified by subject-specific slopes. RESULTS: At baseline and at follow-up, boys tended to be taller, leaner, more aerobically fit, and had greater LVM than girls. Rate of change for these variables was also greater in boys than girls. For example, LVM increased 62% in boys and 48% in girls. At year 5, subjects had advanced at least 1 stage in genital or breast development and over 80% of the subjects were in late- or post-puberty. Significant and strong tracking of heart size (r(s) =.65-.87) was observed. The likelihood that a subject would be in an extreme tertile for heart size at follow-up was approximately doubled if he or she started there at baseline. In boys, baseline FFM explained 54% of the variability in follow-up LVM. Change in aerobic fitness and change in FFM explained 55% of the variability in change in LVM. In girls, baseline aerobic fitness and fatness explained 45% of the variability in follow up LVM. Because FFM did not enter in this model, we constructed an alternative model in which baseline aerobic fitness adjusted for FFM was entered. Using this approach, 43% of the variability in follow-up LVM was explained by baseline FFM, fatness, and adjusted aerobic fitness. Change in FFM explained 58% of the variability in change in LVM. (ABSTRACT TRUNCATED) PMID- 10799628 TI - Ticlopidine plus aspirin for coronary thrombosis in Kawasaki disease. AB - Selective inhibitors of the adenosine 5'-diphosphate pathway of platelet activation have been used rarely in children in the United States. We report the successful use of ticlopidine, together with aspirin, in a 7-month-old infant with Kawasaki disease complicated by a thrombus in a giant coronary aneurysm that failed to resolve with thrombolytic therapy. Kawasaki disease, coronary aneurysms, antithrombotic therapy, ticlopidine, children. PMID- 10799630 TI - False-positive tricyclic antidepressant drug screen results leading to the diagnosis of carbamazepine intoxication. AB - Ingestion of toxic substances is a common problem in pediatrics. When presented with the limited history of an unknown ingestion in a patient with altered mental status, a clinician depends on the physical examination and a toxic screen to determine the ingested substance(s). Some toxic screens yield false-positive or false-negative results that confound identification of ingested toxins. Three cases are presented in which carbamazepine ingestions were identified because of the false-positive tricyclic antidepressant serum toxic screen result in each case. Carbamazepine ingestion is one of the most common pediatric overdoses. Side effects include altered mental status, tachycardia, mydriasis, seizures, coma, and death. Several other substances also cause false-positive tricyclic antidepressant toxic screen results, including certain antipsychotic medications, antihistamines, and the muscle relaxant cyclobenzaprine. Specific tests and drugs causing false-positive results are presented in table form. More modern methods, specifically gas chromatographic-mass spectrometric, are more reliable in distinguishing these drugs. Knowledge of which substances commonly cause false positive results on a given toxic screen can still lead the clinician to the correct diagnosis. tricyclic, carbamazepine, ingestion, intoxication, drug screen. PMID- 10799629 TI - Counseling parents regarding prognosis in autistic spectrum disorder. AB - A triaxial model for autistic spectrum disorder (ASD) is presented, incorporating age, degree of intelligence, and severity of autistic features. As the name implies, ASD can vary in degree of expression from minimal to profound. Furthermore, the symptoms of ASD change in predictable ways with the passage of time. For example, echolalia during early childhood may be replaced by verbal literalism and difficulty with verbal humor during later childhood or adolescence. The prognosis for children with ASD is governed by the joint impact of the degree of expression of ASD and the degree of developmental delay, if any. All combinations of ASD and intellect are possible (ie, severe ASD plus severe mental retardation, severe ASD plus normal general intelligence, and so forth). The relationship among these 3 parameters-severity of ASD, level of general intelligence, and change in symptom expression over time, is represented schematically as a 3-dimensional graph. The utility of this graph as a counseling tool, and as the basis for future research on the prognosis of ASD are discussed. PMID- 10799631 TI - Undiluted albuterol aerosols in the pediatric emergency department. AB - OBJECTIVE: To evaluate the efficacy and efficiency of a short treatment method of administering albuterol aerosols. METHODS: Fifty children 6 to 18 years of age with severe acute asthma (peak flow rates <60% of predicted) were enrolled in a single-blind, controlled trial in an urban pediatric emergency department. Patients were randomized to receive either the study short treatment (3.5 mL of undiluted albuterol nebulized for 20 inhalations) or the control treatment (albuterol diluted [.5 mL] and nebulized in normal saline [3 mL]) every 20 minutes for a total of 3 treatments. Peak flow and spirometric measurements were performed before and after each treatment. RESULTS: There were 25 patients in the study group and 25 in the control group. There were no demographic differences between groups; both had comparable pulmonary function at presentation. The mean forced expiratory volume in 1 second percent predicted improvement between 0 and 60 minutes was 18.8% in the study group and 14.5% in the control group. The total time of treatment delivery for the study group was 6.4 minutes versus 32.7 minutes for the control group. CONCLUSION: Undiluted albuterol short treatments seem to be as effective as standard diluted albuterol in severe asthmatics, while offering the ease and efficiency of shorter treatment administration time. PMID- 10799632 TI - Presentation of low anorectal malformations beyond the neonatal period. AB - OBJECTIVE: Anorectal malformations are usually diagnosed at birth, but some patients have presented to this institution beyond the early newborn period without recognition of their anorectal malformations. To quantify the extent of this problem, we undertook a review of all patients presenting to this hospital with anorectal malformations. METHODS: We reviewed all new cases of anorectal malformations treated at British Columbia's Children's Hospital during the past 11 years. We looked specifically at the time of diagnosis, patient age, sex and mode of presentation, the type of anorectal malformations, and any associated anomalies. RESULTS: One hundred twenty new cases of anorectal malformations were seen here, of whom, 15 patients (9 girls and 6 boys) presented beyond the early newborn period. Of these, 1 male infant was diagnosed at 2 weeks of age and another girl at 14 years of age. The remaining 13 presented between 3 and 11 months of age because of increasing constipation, usually associated with the introduction of solid foods. All had low anorectal malformations. Nine patients had at least 1 other feature of the VACTERL complex. CONCLUSIONS: Most anorectal malformations are identified at birth, but a significant number of the milder lesions may not be recognized until later. Therefore, this condition must be considered in older infants and children presenting with constipation, particularly if they also have cardiac or genitourinary anomalies. constipation, imperforate anus, VACTERL. PMID- 10799633 TI - Coccygeal pits. AB - BACKGROUND: Congenital dermal sinuses represent cutaneous depressions or tracts that are lined by stratified squamous epithelium. They communicate between the surface of the skin and deeper structures and may occur anywhere along the craniospinal axis. These sinuses are thought to result from abnormal separation of the cutaneous and neural ectoderm between the third and fifth week of intrauterine life. They may be often accompanied by other cutaneous stigmata, various dysraphic abnormalities, or intraspinal tumors. In the sacrococcygeal area, cutaneous congenital abnormalities are relatively common. It is estimated that 2% to 4% of children harbor intergluteal dorsal dermal sinuses. These intergluteal sinuses in the perianal region are frequently referred to as pits or dimples. Their cause is considered similar to other congenital dermal sinuses and appears unrelated to acquired pilonidal conditions observed in adults. They may become susceptible to local recurrent infection from trauma or hirsutism. Controversy regarding the evaluation and management of cutaneous defects in the coccygeal region exists. METHODS: Both a literature review and a career review of clinical material were performed. Databases for articles published in English were surveyed for key words relating to coccygeal sinuses using standard computerized search techniques. The medical records of children presenting to our neurosurgical clinic for evaluation of dorsal dermal sinuses were reviewed to identify those with intergluteal sinuses. RESULTS: In the evaluation of reported cases and of our own, we were unable to identify any children with coccygeal sinuses without other cutaneous markers other than hair with findings suggestive of intraspinal communication. CONCLUSIONS: Intergluteal dorsal dermal sinuses are relatively common lesions that frequently come to neurosurgical attention. They do not seem to be associated with significant risk of spinal cord and intraspinal anomalies. Simple intergluteal dorsal dermal sinuses without other cutaneous findings do not require radiographic or surgical evaluation and treatment. If other markers or neurologic symptoms are present, however, radiographic evaluation may be indicated. PMID- 10799634 TI - Familial Mediterranean fever: effects of genotype and ethnicity on inflammatory attacks and amyloidosis. AB - OBJECTIVE: The gene causing familial Mediterranean fever (FMF)-an autosomal recessive disease characterized by recurrent short episodes of fever associated most commonly with peritonitis, pleuritis, and arthritis-has recently been found and several mutations identified. The most severe complication of the disease is amyloidosis, which can lead to renal failure. The aim of this study was to investigate the role of genetic versus nongenetic factors on the phenotype as well as on the development of amyloidosis in FMF in a large and heterogeneous group of patients. METHODOLOGY: We studied 382 patients from 4 ethnic origins living in different environments: North African Jews, other Jews, Turks, Armenians living in the United States, and Armenians from Yerevan, Armenia. Information regarding amyloidosis was available for 371 patients. We examined the association between the mutation M694V and the development of amyloidosis, and we also compared the clinical characteristics of the inflammatory attacks in patients from different ethnic origins, while controlling for the type of mutation. RESULTS: A significant association was found between amyloidosis and the most common mutation in exon 10 of the FMF gene (MEFV), M694V (for M694V homozygotes, relative risk = 1.77; 95% CI = 1.16-2.71). Amyloidosis was present in 44 of 171 homozygous FMF patients (25.7%), in 22 of 143 compound heterozygous FMF patients (15.4%), and in 7 of 57 patients carrying other mutations (12.3%). In homozygotes for M694V who had not been treated with colchicine before 20 years of age, the risk of amyloidosis developing before this age was 61.0%. In our series, there were no cases of amyloidosis in 16 patients carrying the common mutation E148Q. We found that the type and severity of the FMF inflammatory symptoms were associated with both the genotype and the country of residence of the patient. CONCLUSIONS: In the light of the high frequency of amyloidosis in homozygotes for the mutation M694V, colchicine treatment should be given to this group irrespective of the severity of the inflammatory attacks to prevent the development of amyloidosis. Our findings also suggest that factors other than genotype, such as environment or genes other than MEFV, play a role in the determination of the severity of the inflammatory attacks in FMF. amyloidosis, specific mutation, phenotype-genotype correlation, ethnicity. PMID- 10799635 TI - A custodian cured the doctor! PMID- 10799636 TI - FORUM: The Relationship of Ecosystem Management to NEPA and Its Goals. AB - / The National Environmental Policy Act of 1969 (NEPA) was intended to promote a systematic, comprehensive, interdisciplinary approach to planning and decisionmaking, including the integration of the natural and social sciences and the design arts. NEPA critics have cited three key shortcomings in its implementation: (1) a lack of engagement with the NEPA process early in the planning process through interdisciplinary collaboration; (2) a lack of rigorous science and the incorporation of ecological principles and techniques; and (3) a lack of emphasis on the Act's substantive goals and objectives. In recent years and independent of NEPA, a policy of ecosystem management has been developed, which represents a fundamental change from a fragmented, incremental planning and management approach to a holistic, comprehensive, interdisciplinary land and resource management effort. We postulate that by incorporating ecosystem management principles in their planning and decisionmaking, federal agencies can address the shortcomings in NEPA implementation and move closer to NEPA's intent. A case analysis of EISs prepared by the USDA Forest Service before and after adopting an ecosystem management approach supports our hypothesis. PMID- 10799637 TI - PROFILE: Using GIS to Identify Functionally Significant Wetlands in the Northeastern United States. AB - / Of the several automated wetland assessment methods currently available, none are comprehensive in considering all of the primary functions a wetland can perform. We developed a methodology particularly suited to the Northeastern United States that enumerates spatial predictors of wetland function for three primary wetland functions: flood flow alteration, surface water quality improvement, andwildlife habitat. Predictors were derived from several wetland assessment techniques and directly from the literature on wetland structure and function. The methodology was then automated using a Geographic Information System (GIS). The resulting Automated Assessment Method for Northeastern Wetlands (AMNEW) consists of a suite of eight Arc Macro Language (AML) programs that run in the ARC/INFO GRID module. Using remotely sensed land use information and digital elevation models (DEMs), AMNEW produces three separate grids of wetlands that perform each function. The method was tested on four watersheds in Vermont's Lake Champlain Basin. Results and preliminary verification indicate that the method can successfully identify those wetlands in the Northeastern region that have the potential to be functionally important. PMID- 10799638 TI - Demonstration of GIS Capabilities for Fisheries Management Decisions: Analysis of Acquisition Potential Within the Meramec River Basin. AB - / Geographic information systems (GIS) allow users to explore possible spatial relations that may exist within their data. At the Missouri Department of Conservation (MDC), GIS data is being used to help make management decisions. Thirteen geographic data layers of the Meramec River Basin, Missouri, were used to help demonstrate the usefulness of GIS for making fisheries management decisions. The data were used to help identify potential acquisition areas within the Meramec River Basin. The basin was separated into 22 strata based on ecoregion boundaries, watershed boundaries, and stream order. Suitability for acquisition was determined for each stratum based on species richness, habitat characteristics, percent of public land, and number of human impacts, such as gravel and ore mining. Eleven strata scored high enough to be recommended as potential acquisition areas. After further evaluation of the 11 strata, three were chosen as areas where available land and willing sellers should be considered. Four strata needed more sampling before land within them should be considered for acquisition. The final four were considered low priority because there was already a considerable amount of public land present in the strata. The analysis was helpful in allowing managers to focus in on a smaller area for acquisition consideration; 91% of the area was eliminated from the analysis. Instead of having to survey every parcel of land that becomes available, parcels that don't fall within the recommended strata can be eliminated without further investigation. PMID- 10799639 TI - Environmental Protection in the Tomsk Region of the Russian Federation: A Case Study. AB - / Implementation of environmental management in Russia is undergoing rapid changes. Federal responsibility rests with the Russian Federation State Committee for Environmental Protection (RFSCEP) and is delegated at regional level to local State Committees for Environmental Protection (SCEPs). This paper focuses on the functioning of the SCEP for Tomsk oblast' (region) in Siberia, which is strongly committed to forging constructive links with regional government, academia, industry, and environmental NGOs. Considerable difficulties exist for SCEPs in Russia, however, and prominent among these are (1) a rigidly vertical civil service structure, with separate organs having responsibility for different natural resources, persisting from Soviet times, which hinders effective intergrated and holistic environmental management; and (2) a lack of open access to environmental information from military and quasi-military sites. PMID- 10799640 TI - A Mediation Case for Resolving the Energy and Environment Dispute at Aliaga Izmir, Turkey. AB - / Aliaga town, located 50 km north of Izmir, Turkey, is facing serious air, water, and soil pollution problems of industrial origin. The town has had a widespread public reaction against the estimated environmental effects of a 500 MW power plant originally to be built by a private international company during the first half of the 1990s. This project was rejected by court order at that time, but recently a number of new power projects emerged, and overall environmental burdens had to be reconsidered.A mediation exercise to resolve the ongoing dispute against these power plant projects at Aliaga was recommended and participated in by the author in 1997. In this article the basis of the continuing environmental concern about the feared impacts of the new power plants, procedure, and results of this mediation are mentioned. The basis of the "energy versus environment" dispute in Aliaga are introduced. Mediation exercise and its end results have been criticized. PMID- 10799641 TI - A Multimethodological Approach for the Sustainable Management of Perifluvial Wetlands of the Po River (Italy). AB - / Marginal aquatic systems (wetlands) of the Po River (Italy) have become the target of a renewed interest because of their value for recreation, natural reserves, and deposits of sand. To preserve these sites, wise management must be the objective of local administrations. In this paper a strategy for the sustainable use of 11 wetlands is presented. It uses simple economic analysis and multiple criteria techniques and provides suggestions to promote sustainability in terms of conservation of natural resources, economic self-sufficiency, and minimization of potential conflicts about the use of the wetlands. In the understanding that sustainability is framed in a long-term perspective, stability analysis is also considered and performed by means of loop analysis, a qualitative technique. Conditions for stability are then discussed about management opportunities. PMID- 10799642 TI - RESEARCH: Influence of Social, Biophysical, and Managerial Conditions on Tourism Experiences Within the Great Barrier Reef World Heritage Area. AB - / Managing protected areas involves balancing the enjoyment of visitors with the protection of a variety of cultural and biophysical resources. Tourism pressures in the Great Barrier Reef World Heritage Area (GBRWHA) are creating concerns about how to strike this balance in a marine environment. Terrestrial-based research has led to conceptual planning and management frameworks that address issues of human use and resource protection. The limits of acceptable change (LAC) framework was used as a conceptual basis for a study of snorkeling at reef sites in the GBRWHA. The intent was to determine if different settings existed among tourism operators traveling to the reef and, if so, to identify specific conditions relating to those settings. Snorkelers (N = 1475) traveling with tourism operations of different sizes who traveled to different sites completed surveys. Results indicated that snorkelers who traveled with larger operations (more people and infrastructure) differed from those traveling with smaller operations (few people and little on-site infrastructure) on benefits received and in the way that specific conditions influenced their enjoyment. Benefits related to nature, escape, and family helped to define reef experiences. Conditions related to coral, fish, and operator staff had a positive influence on the enjoyment of most visitors but, number of people on the trip and site infrastructure may have the greatest potential as setting indicators. Data support the potential usefulness of visitor input in applying the LAC concept to a marine environment where tourism and recreational uses are rapidly changing. PMID- 10799643 TI - Erosion Control at Construction Sites: The Science-Policy Gap. AB - / To test the effectiveness of Michigan's soil erosion control law, 30 construction sites were evaluated in the east-central part of the state. The analytical framework lumped nine best management practices (BMPs) most closely related to the law into three categories: slope stabilization, soil stabilization, and water management. All sites were in the land clearing or foundation/framing stage of construction and were evaluated within 2 days after a rainfall event. Only four of the sites performed above the mean of the scoring scale, with the categorical scoring of BMPs indicating the worst performance for slope stabilization measures. The poor results suggest a failure to integrate scientific knowledge of erosion control with policy. A fundamental problem is the lack of basic site data on soil, topography, and hydrology, resulting in the incorrect application of BMPs, such as staging, filter fences, and berms. The current institutional framework for soil erosion control also provides disincentives to mitigate local erosion problems. PMID- 10799644 TI - ENVIRONMENTAL AUDITING: Chemical Characteristics of Urban Stormwater Sediments and Implications for Environmental Management, Maricopa County, Arizona. AB - / Investigations of the chemical characteristics of urban stormwater sediments in the rapidly growing Phoenix metropolitan area of Maricopa County, Arizona, showed that the inorganic component of these sediments generally reflects geologic background values. Some concentrations of metals were above background values, especially cadmium, copper, lead, and zinc, indicating an anthropogenic contribution of these elements to the sediment chemistry. Concentrations, however, were not at levels that would require soil remediation according to guidelines of the U.S. Environmental Protection Agency. Arsenic concentrations generally were above recommended values for remediation at a few sites, but these concentrations seem to reflect geologic rather than anthropogenic factors. Several organochlorine compounds no longer in use were ubiquitous in the Phoenix area, although concentrations generally were low. Chlordane, DDT and its decay products DDE and DDD, dieldrin, toxaphene, and PCBs were found at almost all sites sampled, although some of the pesticides in which these compounds are found have been banned for almost 30 years. A few sites showed exceptionally high concentrations of organochlorine compounds.On the basis of published guidelines, urban stormwater sediments do not appear to constitute a major regional environmental problem with respect to the chemical characteristics investigated here. At individual sites, high concentrations of organic compounds-chlordane, dieldrin, PCBs, and toxaphene-may require some attention. The possible environmental hazard presented by low-level organochlorine contamination is not addressed in this paper; however, high levels of toxicity in urban sediments are difficult to explain. Sediment toxicity varied significantly with time, which indicates that these tests should be evaluated carefully before they are used for management decisions. PMID- 10799645 TI - Mechanism of action in thalidomide teratogenesis. AB - In this commentary, we describe a model to explain the mechanism of the embryopathy of thalidomide. We propose that thalidomide affects the following pathway during development: insulin-like growth factor I (IGF-I) and fibroblast growth factor 2 (FGF-2) stimulation of the transcription of alphav and beta3 integrin subunit genes. The resulting alphavbeta3 integrin dimer stimulates angiogenesis in the developing limb bud, which promotes outgrowth of the bud. The promoters of the IGF-I and FGF-2 genes, the genes for their binding proteins and receptors, as well as the alphav and beta3 genes, lack typical TATA boxes, but instead contain multiple GC boxes (GGGCGG). Thalidomide, or a breakdown product of thalidomide, specifically binds to these GC promoter sites, decreasing transcription efficiency of the associated genes. A cumulative decrease interferes with normal angiogenesis, which results in truncation of the limb. Intercalation into G-rich promoter regions of DNA may explain why certain thalidomide analogs are not teratogenic while retaining their anti-tumor necrosis factor-alpha (TNF-alpha) activity, and suggests that we look elsewhere to explain the action of thalidomide on TNF-alpha. On the other hand, the anti-cancer action of thalidomide may be based on its antiangiogenic action, resulting from specific DNA intercalation. The tissue specificity of thalidomide and its effect against only certain neoplasias may be explained by the fact that various developing tissues and neoplasias depend on different angiogenesis or vasculogenesis pathways, only some of which are thalidomide-sensitive. PMID- 10799646 TI - Regional and cellular induction of nicotine-metabolizing CYP2B1 in rat brain by chronic nicotine treatment. AB - In the rat, nicotine is metabolized to cotinine primarily by hepatic cytochrome P450 (CYP) 2B1. This enzyme is also found in other organs such as the lung and the brain. Hepatic nicotine metabolism is unaltered after nicotine exposure; however, nicotine may regulate CYP2B1 in other tissues. We hypothesized that nicotine induces its own metabolism in brain by increasing CYP2B1. Male rats were treated with nicotine (0.0, 0.1, 0.3, or 1.0 mg base/kg in saline) s.c. daily for 7 days. CYP2B1 mRNA and protein were assayed in the brain and liver by reverse transcriptase-polymerase chain reaction (RT-PCR), immunoblotting, and immunocytochemistry. In control rats, CYP2B1 mRNA and protein expression were brain region- and cell-specific. CYP2B1 was not induced in the liver, but CYP2B1 mRNA and protein showed dose-dependent, region- and cell-specific patterns of induction across brain regions. At 1.0 mg nicotine/kg, the largest increase in protein was in the brain stem (5.8-fold, P < 0.05) with a corresponding increase in CYP2B1 mRNA (7.6-fold, P < 0.05). Induction of CYP2B1 was also observed in the frontal cortex, striatum, and olfactory tubercle. Immunocytochemistry showed that induction was restricted principally to neurons. These data indicate that nicotine may alter its own metabolism in the brain through transcriptional regulation, perhaps contributing to central tolerance to the effects of nicotine. CYP2B1 and its human homologue CYP2B6 also activate tobacco smoke procarcinogens such as NNK [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone]. Highly localized increases in CYP2B could result in increased mutagenesis. These data suggest roles for nicotine-induced CYP2B in central metabolic tolerance, nicotine-induced neurotoxicity, neuroplasticity, and carcinogenesis. PMID- 10799647 TI - Protein dephosphorylation rates in myocytes after isoproterenol withdrawal. AB - Dephosphorylation of substrates for cyclic AMP-dependent protein kinase is essential for reversing the effects of this enzyme. It has been proposed that the relevant phosphatase(s) is stimulated by muscarinic cholinergic agonists, thereby accentuating cholinergic antagonism of beta-adrenergic agonists in the heart. To test this hypothesis, dephosphorylation of the three major substrates of cardiac cyclic AMP-dependent protein kinase (phospholamban, troponin-I, and C-protein) was examined. In isolated myocytes, isoproterenol caused concentration-dependent phosphorylation of these three proteins. Simultaneous exposure to acetylcholine with the isoproterenol caused a rightward shift in the concentration-response curve that was similar for protein phosphorylation in myocytes and for the inotropic response of the intact heart. The addition of propranolol after exposure to isoproterenol resulted in protein dephosphorylation, the onset of which was accelerated by acetylcholine. However, acetylcholine did not affect the rate of dephosphorylation for any of the proteins, indicating that phosphatase activity in cardiac muscle is not enhanced by acetylcholine. PMID- 10799648 TI - Shifts in the affinity distribution of one class of seven-transmembrane receptors by activation of a separate class of seven-transmembrane receptors. AB - We have demonstrated previously that activation of thrombin receptors causes increased Galpha(q) coupling to thromboxane A(2) receptors and increased thromboxane A(2) receptor ligand affinity. These results led to the hypothesis that thrombin receptor activation stimulates Galpha(q) redistribution to thromboxane A(2) receptors, thereby shifting them to a higher affinity state. The present study investigated three questions regarding this inter-receptor signaling phenomenon: (i) does activation of thrombin receptors cause a redistribution of thromboxane A(2) receptor subpopulations; (ii) does inter receptor signaling require that participating receptors couple to the same family of G-protein alpha-subunits; and (iii) does inter-receptor signaling occur in cell types other than platelets? It was found that thrombin receptor activation caused a shift in the thromboxane A(2) receptor binding data from a one-site model to a two-site model (K(i) = 0.5 microM vs K(i) = 10 nM and 1.1 microM for the antagonist 4-[2-[[(4-chlorophenyl)sulfonyl]amino]ethyl]benzeneacetic acid (BM13. 505) and K(i) = 2.5 microM vs K(i) = 29.5 nM and 2.6 microM for the agonist 9,11-dideoxy-9alpha,11alpha-methanoepoxy prostaglandin F(2alpha) (U46619). It also was found that activation of prostaglandin D(2) receptors also caused a shift of prostacyclin receptor binding data from a one-site model (IC(50) = 10.1 nM) to a two-site model (IC(50) = 3.3 and 12.5 nM). The physiological manifestation of this inter-receptor signaling between prostacyclin and prostaglandin D(2) receptors was a synergistic inhibition of human platelet aggregation. Finally, the present results established that activation of endothelial cell thrombin receptors shifts thromboxane A(2) receptor affinity from K(i) = 0.8 microM (control) to K(i) = 0.2 microM (thrombin receptor activating peptide), indicating that cells other than platelets have the capability to signal between seven-transmembrane receptors. PMID- 10799649 TI - alpha(2A)-adrenoceptor: G(alphai1) protein-mediated pertussis toxin-resistant attenuation of G(s) coupling to the cyclic AMP pathway. AB - Fusion proteins were constructed between a recombinant human alpha(2A) adrenoceptor and either a rat wild-type G(alphai1) or putative pertussis toxin resistant form of the G(alphai1) protein (G(alphai1)Cys(351)Gly). [(3)H]2-[2-(2 Methoxy-1, 4-benzodioxanyl)]imidazoline hydrochloride (RX 821002) saturation binding experiments demonstrated that both fusion proteins were expressed at a similar level as the alpha(2A)-adrenoceptor co-expressed with either a wild-type G(alphai1) or mutant G(alphai1)Cys(351)Gly protein in COS-7 cells, and displayed a ligand binding profile similar to that for the alpha(2A)-adrenoceptor protein. In alpha(2A)-adrenoceptor-transfected COS-7 cells, 5-bromo-6-(2-imidazolin-2-yl amino) quinoxaline tartrate (brimonidine, 10 microM) induced stimulation (151 +/- 28%) of adenosine 3',5'-cyclic monophosphate (cAMP) formation which was prevented by cholera toxin treatment, demonstrating a direct coupling of the alpha(2A) adrenoceptor to an endogenous G(alphas) protein in COS-7 cells. Expression of either the wild-type G(alphai1) or mutant G(alphai1)Cys(351)Gly protein in co expression or fusion with the alpha(2A)-adrenoceptor in COS-7 cells suppressed the brimonidine-induced stimulation of cAMP formation, both in the presence and absence of pertussis toxin pretreatment. Hence, the G(alphai1) protein apparently blocks the G(s)-coupled alpha(2A)-adrenoceptor-mediated pathway in a pertussis toxin-non-sensitive way. PMID- 10799650 TI - Clavilactones, a novel class of tyrosine kinase inhibitors of fungal origin. AB - Targeting of deregulated protein tyrosine kinases has been proposed as a new approach in the therapeutic intervention against pathological processes including proliferative disorders and cancer. Using a screening approach based on a comparative evaluation of antiproliferative effects in a panel of tumor cells with differential expression of protein tyrosine kinases, three benzoquinoid macrolidic fungal metabolites produced by Clitocybe clavipes, clavilactones A, B, and D (CA, CB, and CD) and two semisynthetic derivatives of these products, diacetyl-CA and dimethyl-CA, were identified as inhibitors of protein tyrosine kinases. Naturally occurring CA, CB, and CD showed inhibitory activity in kinase assays against the Ret/ptc1 and epidermal growth factor receptor (EGF-R) tyrosine kinases, while being less effective against the v-Abl tyrosine kinase and p34(cdc2) serine/threonine kinase (IC(50) 2.8, 5.5, 81.3, and 128 microM respectively, for the most potent compound CD). CB was shown to be a non competitive inhibitor of EGF-R with respect to ATP or poly(Glu(6)Ala(3)Tyr). CD also preferentially inhibited the growth of A431 cells, which overexpress a constitutively active EGF-R, as opposed to IGROV-1 and SKOV-3 cells, which express low levels of the receptor. Further, EGF-R was shown to be a target for clavilactones in A431 cells, since EGF-induced receptor autophosphorylation was inhibited in the presence of CB, CD, and diacetyl-CA. Both CD and diacetyl-CA displayed weak activity when administered daily (i.p.) to mice bearing ascitic A431 tumor. These findings indicate that clavilactones represent the prototypes of a new structural class of tyrosine kinase inhibitors deserving further investigation. PMID- 10799651 TI - Inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-stimulated Cyp1a1 promoter activity by hypoxic agents. AB - Since hypoxia-inducible factor-1alpha (HIF-1alpha) and the arylhydrocarbon receptor (AhR) shared the AhR nuclear translocator (Arnt) for hypoxia- and AhR mediated signaling, respectively, it was possible to establish the hypothesis that hypoxia could regulate cytochrome P450 1a1 (Cyp1a1) expression. In order to test this hypothesis, we undertook to examine the effect of hypoxia on Cyp1a1 transcription in Hepa-I cells. Mouse Cyp1a1 5'-flanking DNA, 1.6 kb was cloned into pGL3 expression vector in order to construct pmCyp1a1-Luc. Hepa-I cells were transfected with pmCyp1a1-Luc and treated with 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) in the presence or absence of various hypoxic agents such as 1-100 microM cobalt chloride, 1-100 microM picolinic acid, and 1-100 microM desferrioxamine. Luciferase activity of the reporter gene was measured from pmCyp1a1-Luc-transfected Hepa-I cell lysate which contains 2 microgram total protein using luciferin as a substrate. Hypoxic agents such as cobalt chloride, picolinic acid, and desferrioxamine showed inhibition of luciferase activity that was induced by 1-nM TCDD treatment in a dose-and time-dependent manner. Concomitant treatment of 150 microM ferrous sulfate with 1-100 microM desferrioxamine or 1-100 microM picolinic acid recovered luciferase activity from that inhibited by hypoxic agents or induced by TCDD. These data demonstrated that iron-chelating and hypoxic agents inhibited dioxin-induced Cyp1a1 transcription in Hepa-I cells. Thus, we might suggest that hypoxia inhibits TCDD-induced Cyp1a1 expression due to the competition between HIF-1alpha and the AhR for the Arnt in Hepa-I cells. PMID- 10799652 TI - Development of resistance to glutathione depletion-induced cell death in CC531 colon carcinoma cells: association with increased expression of bcl-2. AB - The glutathione (GSH) level of CC531 rat colorectal cancer cells is readily decreased by exposure to buthionine sulfoximine (BSO), an inhibitor of GSH synthesis; at 25 microM BSO, these cells died in a non-apoptotic fashion. By continuous exposure of CC531 cells to increasing concentrations of BSO, we obtained a BSO-resistant cell line (CCBR25) that was 50 times more resistant to BSO than the parental cell line. Whereas the GSH content of CCBR25 and CC531 cells was similar, the former contained a much higher level of the Bcl-2 protein. After stable transfection of CC531 cells with the human bcl-2 gene, the resulting Bcl-2-overexpressing cell line appeared to be 9 times more resistant to BSO than the parental cell line. These findings suggest that the Bcl-2 protein offers resistance against the cytotoxic effect of severe GSH depletion. PMID- 10799653 TI - Identification of the human cytochromes P450 involved in the oxidative metabolism of "Ecstasy"-related designer drugs. AB - The human cytochrome P450 (CYP) isozymes catalyzing the oxidative metabolism of the widely abused amphetamine derivatives MDMA (N-methyl-3,4 methylenedioxyamphetamine, "Ecstasy"), MDE (N-ethyl-3, 4 methylenedioxyamphetamine, "Eve"), and MDA (3, 4-methylenedioxyamphetamine) were identified. Using a simplified non-extractive reversed-phase HPLC assay with fluorescence detection, biphasic Michaelis-Menten kinetics were obtained for formation of all three dihydroxyamphetamines in liver microsomes from a CYP2D6 extensive metabolizer subject. In contrast, no low K(m) component was detectable in microsomes from a poor metabolizer subject. Additional specific probes for CYP2D6 further confirmed this isozyme as the exclusive low K(m) component for demethylenation. P450-selective inhibitors applied to CYP2D6-inhibited microsomes and activity measurements in a series of recombinant P450s suggested CYP1A2 as the major high K(m) component with contributions by CYP2B6 and CYP3A4. Moreover, the relative CYP1A2 content of a panel of 12 human livers was weakly but significantly correlated to the high K(m) demethylenase activity (Spearman rank correlation coefficient [r(s)] = 0.58; P < 0.05). Microsomal maximal velocities for N-dealkylation were at least 7-fold lower than for demethylenation and were characterized by apparently monophasic kinetics. The most important isozyme for this reaction appeared to be CYP2B6, the microsomal content of which was found to be strongly correlated to N-deethylation of MDE (r(s) = 0.90; P < 0.001). We conclude that, in addition to CP2D6 as the sole high-affinity demethylenase, several other P450 isozymes have the capacity to contribute to microsomal oxidative metabolism of methylenedioxyamphetamines. This may be of particular importance in individuals genetically lacking functional CYP2D6. PMID- 10799654 TI - Increased cytotoxicity of cadmium in fibroblasts lacking c-fos. AB - Cadmium has been known to induce the expression of the c-fos gene in various cell types including fibroblasts. To clarify the biological significance of c-fos induction by cadmium, mouse 3T3-like fibroblasts lacking c-fos were exposed to cadmium, and the resultant cellular damage was assayed by WST-8 (4-[3-(2-methoxy 4-nitrophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio] -1, 3-benzene disulfonate sodium salt) conversion, trypan blue exclusion, or lactate dehydrogenase leakage. The c-fos-deficient cells (f1 and f10) were affected more severely than the wild type cells (NIH 3T3 and f20) with respect to both cell growth and cellular damage following exposure to 10 or 20 microM cadmium chloride. These results suggest that c-fos may play a protective role against the cytotoxic effects of cadmium at least in these 3T3-like fibroblasts. PMID- 10799655 TI - Stimulatory effect of curcumin on osteoclast apoptosis. AB - Curcumin is a potent inhibitor of the transcriptional factors activator protein-1 and nuclear factor-kappaB. Since transcriptional factors may play a functional role in the survival of osteoclasts, it was of interest to us to examine the effect of curcumin on osteoclast apoptosis. We observed that curcumin is a potent stimulator of this process in rabbit osteoclasts, as evidenced by morphological changes in nuclei and DNA fragmentation as criteria of apoptosis. The curcumin stimulation of the osteoclast apoptosis was dose-and treatment time-dependent. In addition, curcumin dramatically inhibited osteoclastic bone resorption, supporting our data that curcumin is a potent stimulator of osteoclast apoptosis. PMID- 10799656 TI - Expression of human mitochondrial thymidine kinase in Escherichia coli: correlation between the enzymatic activity of pyrimidine nucleoside analogues and their inhibitory effect on bacterial growth. AB - Mitochondrial thymidine kinase (TK2) phosphorylates pyrimidine nucleosides to monophosphates and is expressed constitutively through the cell cycle in all cells. Because of the overlap of its substrate specificity with that of the cytosolic thymidine kinase (TK1) and deoxycytidine kinase (dCK), it has been difficult to determine the role of TK2 in activating nucleosides used in chemotherapy. In this report, we described the construction of a recombinant Escherichia coli strain which could be used to test if TK2 activity is limiting for the toxicity of nucleosides. Enzymes of bacterial origin which are involved in thymidine and deoxyuridine anabolism and catabolism were eliminated, and the cDNA for human TK2 was introduced. In the crude extract of the engineered E. coli, the level of thymidine kinase was, after induction of TK2 expression, several hundred fold higher than in the control strain. Several pharmacologically interesting nucleoside analogues, including 3'-azidothymidine, 2',3'-didehydro 2',3'-dideoxythymidine, and 2', 3'-dideoxy-beta-L-3'-thiacytidine, were tested for their effects on the growth of this recombinant strain. For a comparison, the phosphorylation of these compounds was determined with purified recombinant TK1, TK2, and dCK. A correlation was observed between the phosphorylation of several of these compounds by TK2 and their effects on bacterial growth. These results demonstrate that activation of growth-inhibiting pyrimidine nucleosides can be catalyzed by TK2, and together with recombinant E. coli strains expressing other cellular nucleoside kinases, this whole-cell bacterial system may serve as a tool to predict the efficacy and side effects of chemotherapeutic nucleosides. PMID- 10799657 TI - Protective effect of L-deprenyl against apoptosis induced by okadaic acid in cultured neuronal cells. AB - L-Deprenyl, an irreversible MAO-B (monoamine oxidase B, EC 1.4.3.4) inhibitor, is used for the treatment of Parkinson's disease and to delay the progression of Alzheimer's disease. L-Deprenyl also exhibits protective effects against neuronal apoptosis which are independent of its ability to inhibit MAO-B. The purpose of this study was to compare the antiapoptotic efficacy of L-deprenyl against different types of apoptotic inducers in three neuronal cell culture models. The level of apoptosis was quantified by measuring the activation of caspase-3 enzyme, which is the main apoptotic executioner in neuronal cells. MTT [3-(4,5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] and LDH (lactate dehydrogenase, EC 1. 1.1.27) assays were used to demonstrate the cytotoxic response of apoptotic treatments. Our results showed that okadaic acid, an inhibitor of protein phosphatase 1 and 2A, induced a prominent increase in caspase-3 activity both in cultured hippocampal and cerebellar granule neurons as well as in Neuro-2a neuroblastoma cells. Interestingly, L-deprenyl offered a significant protection against the apoptotic response induced by okadaic acid in all three neuronal models. The best protection appeared at the concentration level of 10(-9) M. L-Deprenyl also provided a protection against apoptosis after AraC (cytosine beta-D-arabinoside) treatment in hippocampal neurons and Neuro-2a cells and after etoposide treatment in Neuro-2a cells. However, L-deprenyl did not offer any protection against apoptosis caused by serum withdrawal or potassium deprivation. Okadaic acid treatment in vivo is known to induce an Alzheimer's type of hyperphosphorylation of tau protein, formation of beta amyloid plaques, and a severe memory impairment. Our results show that the okadaic acid model provides a promising tool to study the molecular basis of Alzheimer's disease and to screen the neuroprotective capacity of L-deprenyl derivatives. PMID- 10799658 TI - New vitamin D receptor agonists with decreased metabolic stability. AB - The aim of the study was the development of vitamin D receptor agonists with decreased metabolic stability for the topical treatment of psoriasis and related hyperproliferative skin diseases. Calcitriol analogues 1, 2, 3, all of which contain modifications in the side chain, were synthesized. The obtained analogues were full agonists when the induction of CD14 expression in HL-60 cells, the induction of 5-lipoxygenase activity in Mono Mac 6 cells, and the inhibition of phytohemagglutinin (PHA)-stimulated lymphocyte proliferation were studied. The EC(50) value of the most active compound 1 was 1.2 nM in the CD14 assay and 1 nM in the 5-lipoxygenase assay, whereas calcitriol gave EC(50) values in these assays of 3.7 and 9 nM, respectively. In the lymphocyte proliferation assay, compound 1 and calcitriol had IC(50) values of 0.3 and 2.8 nM, respectively. All three compounds had receptor binding affinities similar to that of calcitriol. The compounds showed a decreased metabolic stability in rat liver homogenates and had a 50-fold lower affinity for the vitamin D-binding protein than calcitriol, which suggests that calcitriol analogues are metabolized more rapidly after systemic uptake or application. When injected into rats, the analogues displayed an approximately 100-fold lower hypercalcemic effect than calcitriol. In summary, our study presents three new and potent vitamin D receptor agonists with interesting profiles for development as antipsoriatic drugs. PMID- 10799659 TI - Calcitonin gene-related peptide-dependent vascular relaxation of rat aorta. An additional mechanism for nitroglycerin. AB - We investigated the involvement of calcitonin gene-related peptide (CGRP) in the vasodilatory mechanism of action of nitric oxide (NO) donors. The functional role of CGRP in NO donor-induced vasodilation of isolated rat aortic rings was determined by incubating these drugs with and without CGRP(8-37), a selective CGRP receptor antagonist. CGRP(8-37) (0.63 microM) induced rightward shifts in the vasodilatory concentration-response curves for nitroglycerin (NTG), Piloty's acid (PA), and SIN-1 (linsidomine). The EC(50) values for NTG, PA, and SIN-1 were increased by 8.3-, 5.2-, and 2.3-fold, respectively (P < 0.05). The release of CGRP from rat aorta in response to NTG and PA was measured specifically by radioimmunoassay. Thirty-minute incubations of NTG or PA with rat aorta induced 189.5 and 214.6% increases, respectively, in CGRP release when compared with the control (P < 0.05). The concentration-response curves of sodium nitroprusside (SNP), S-nitroso-acetylpenicillamine (SNAP), tetranitromethane (TNM), diethylamine NO complex (DEA-NO), and diethylenetriamine/nitric oxide adduct (DETA NONOate) were not inhibited significantly by CGRP(8-37) co-incubation (P 0.05). NO donors also were incubated with aortic strips, and NTG and PA alone induced significant formation of hydroxylamine, a NO(-) metabolite (232.4 and 364.9%, respectively, P < 0.05). These results indicate that only NTG and PA, and to a lesser extent SIN-1, stimulate the release of CGRP from the rat aorta, which subsequently contributes to the vasodilatory activity of these agents. The hydroxylamine formation suggests a possible link between NO(-) generation and CGRP release from the vascular wall. PMID- 10799661 TI - The effects of bile acids on beta-adrenoceptors, fluidity, and the extent of lipid peroxidation in rat cardiac membranes. AB - Bile acids have been proposed as a causative factor for the cardiomyopathy of cholestatic liver disease, since they cause negative inotropism and chronotropism and attenuate cardiac responsiveness to sympathetic stimulation. Bile acids can also modify membrane fluidity and generate reactive oxygen species (ROS). The effects of 10(-6)-10(-3) M deoxycholic acid (DCA) and chenodeoxycholic acid (CDCA) and their taurine conjugates, TDCA and TCDCA, on (1) the binding characteristics of beta-adrenoceptors, (2) membrane fluidity, and (3) the extent of lipid peroxidation in rat cardiac membranes were assessed. The results were compared to the effects of the oxidant, 10(-4)-10(-3) M hydrogen peroxide (H(2)O(2)), and the membrane-fluidizing compound, 5 x 10(-5) M 2-(2 methoxyethoxy)ethyl 8-(cis-2-n-octylcyclopropyl)octanoate (A(2)C). Cardiac beta adrenoceptor density alone was reduced at 10(-4) M bile acid concentration while, at 10(-3) M bile acids, reductions in both receptor density and affinity were seen. At 10(-4) M H(2)O(2), receptor number and affinity were reduced, whereas A(2)C increased receptor affinity without affecting receptor density. Bile acids (10(-3) M) and 10(-4) M H(2)O(2) reduced membrane fluidity. H(2)O(2) caused a concentration-dependent increase in the extent of lipid peroxidation, whereas the bile acids and A(2)C had no effect. Bile acids (10(-4) M) reduced beta adrenoceptor density in the absence of variations in membrane fluidity and in the extent of membrane lipid peroxidation. This result suggests that bile acids, at concentrations equivalent to the plasma/serum total or estimated free bile acid concentration, may have a possible role in the etiology of cardiomyopathy of cholestatic liver disease. At 10(-3) M bile acid concentration, beta-adrenoceptor number and affinity were adversely affected, accompanied by a decrease in membrane fluidity but without any significant increase in the extent of membrane lipid peroxidation. Although cardiac beta-adrenoceptor density and affinity and membrane fluidity were adversely affected by bile acids, the relevance of these findings to our understanding of the etiological basis of hepatic cardiomyopathy is questionable, since such concentrations exceeded the highest concentrations seen in the plasma and/or tissues of patients with cholestatic liver disease. PMID- 10799660 TI - Characterization of phentermine and related compounds as monoamine oxidase (MAO) inhibitors. AB - Phentermine was shown in the 1970s to inhibit the metabolism of serotonin by monoamine oxidase (MAO), but never was labeled as an MAO inhibitor; hence, it was widely used in combination with fenfluramine, and continues to be used, in violation of their labels, with other serotonin uptake blockers. We examined the effects of phentermine and several other unlabeled MAO inhibitors on MAO activities in rat lung, brain, and liver, and also the interactions of such drugs when administered together. Rat tissues were assayed for MAO-A and -B, using serotonin and beta-phenylethylamine as substrates. Phentermine inhibited serotonin-metabolizing (MAO-A) activity in all three tissues with K(i) values of 85-88 microM. These potencies were similar to those of the antidepressant MAO inhibitors iproniazid and moclobemide. When phentermine was mixed with other unlabeled reversible MAO inhibitors (e.g. pseudoephedrine, ephedrine, norephedrine; estradiol benzoate), the degree of MAO inhibition was additive. The cardiac valvular lesions and primary pulmonary hypertension that have been reported to be associated with fenfluramine-phentermine use may have resulted from the intermittent concurrent blockage of both serotonin uptake and metabolism. PMID- 10799662 TI - Astrocytic hypertrophy and altered GFAP degradation with age in subcortical white matter of the rhesus monkey. AB - Reactive astrocytosis is a well known phenomenon that occurs in the normal aging process of the brain. While many studies indicate astrocytic hypertrophy and glial fibrillary acidic protein (GFAP) content increase with age in the hippocampal formation of certain animal models, it is unclear whether these findings are generalizable to the primate and to other areas of the brain. In this study, we quantitatively assessed age-related changes in astrocytic cell size and density in a rhesus monkey model of normal aging. By GFAP immunohistochemistry, we observed an increase in GFAP(+) cell size but not density in all subcortical white matter areas of the frontal, temporal, and parietal cortices. No significant increases in astrocyte hypertrophy were observed in any gray matter area examined. In addition, Western blotting experiments showed increases in total and degraded GFAP content with age, suggesting altered degradation and possibly production of GFAP occur with age. PMID- 10799664 TI - Differential profiles of nitric oxide and norepinephrine releases in the paraventricular nucleus region in response to mild footshock in rats. AB - The purpose of this study was to determine whether the application of mild intermittent footshock stress can cause changes in the nitric oxide (NO) and norepinephrine (NE) releases in the hypothalamic paraventricular nucleus (PVN) region and medial prefrontal cortex (mPFC). Extracellular levels of NO metabolites and NE in the PVN region and mPFC were determined using an in vivo brain microdialysis technique in conscious rats. In the PVN region, we demonstrated that perfusion of N-methyl-D-aspartate through a microdialysis probe resulted in a dose-dependent increase in NO metabolite levels, whereas intraperitoneal administration of N(G)-nitro-L-arginine methyl ester produced a dose-dependent reduction in the levels of NO metabolites. The levels of NO metabolites in the PVN region increased after intraperitoneal administration of interleukin-1beta in a dose-dependent manner, as we previously reported. This increase in NO metabolite levels was abolished 60 min after systemic administration of N(G)-nitro-L-arginine methyl ester compared to the vehicle treated control group. Twenty minutes of intermittent footshock induced NE release but did not induce NO release in the PVN region. On the contrary, in the mPFC, 20 min of intermittent footshock induced both NO and NE releases. The present results reveal different patterns and time courses in NO and NE releases between the PVN region and the mPFC in response to mild intermittent footshock stress. These findings are likely to have helpful suggestions for our understanding of the hypothalamic-pituitary-adrenal axis and the limbic forebrain system response to different kinds of stress. PMID- 10799663 TI - ARC POMC mRNA and PVN alpha-MSH are lower in obese relative to lean zucker rats. AB - Effects of obesity on gene expression for opioid peptides and neuropeptide-Y (NPY) in the arcuate nucleus (ARC), and on opioid peptides and alpha-melanocyte stimulating hormone (alpha-MSH) in the paraventricular nucleus (PVN) were examined in obese Zucker rats (18 weeks old). Obese Zucker rats are insulin resistant, diabetic and hyperleptinemic as indicated by high serum glucose, insulin and leptin levels. ARC proOpiomelanocortin (POMC) mRNA levels were significantly lower in the obese relative to lean Zucker rats and ARC proNeuropeptide Y (proNPY) mRNA levels were higher (P<0.05). There were no differences in proDynorphin and proEnkephalin mRNA levels in the ARC (0.05). Obese Zucker rats had lower alpha-MSH and dynorphin A(1-17) peptide levels in the paraventricular nucleus (PVN) (P<0.05), but did not have lower PVN beta-endorphin peptide levels (0.05). The decrease in POMC in the ARC and decrease in alpha-MSH in the PVN seen in the obese Zucker rat in the present study suggest that reduced activity of the melanocortin system in the ARC to PVN pathway may contribute to the related hyperphagia. Reduced activity of the melanocortin system in the ARC to PVN pathway may be due to a disturbance of leptin signaling coupling to POMC. PMID- 10799665 TI - alpha(2)-adrenergic receptors are not required for central anti-hypertensive action of moxonidine in mice. AB - In the mouse medulla oblongata, we characterized binding properties and functional responses of two recognition sites for imidazoline compounds: I(1) imidazoline and alpha(2)-adrenergic receptors. The mouse medulla expresses a higher density of I(1)-receptors than in the rat, whereas alpha(2)-receptor densities were similar between the two species. In anesthetized, ventilated and paralyzed mice, we tested the hypotensive actions of the I(1)/alpha(2) agonist moxonidine, determined its central site of its actions, and the relative roles of I(1) and alpha(2)-receptors. Experiments were performed in C(57)Bl(6) wild type and alpha(2A)-adrenergic receptor deficient mice. In both types of mice, neuronal activation within the rostral ventrolateral medulla (RVLM) region by glutamate microinjection elicited increases in arterial pressure. Moxonidine (0.5 nmol/site/10 nl) microinjected bilaterally into this vasopressor region decreased arterial pressure by 30% and heart rate by 11% in wild type mice. Efaroxan, the I(1)/alpha(2) antagonist (0.4 nmol) when microinjected into the RVLM elevated blood pressure itself and abolished the action of moxonidine, whereas alpha(2) blockade with SK&F 86466 had no significant effect on blood pressure and did not attenuate moxonidine's effect. To more definitively test the role of alpha(2) adrenergic receptors in the action of moxonidine, moxonidine was microinjected into the RVLM of alpha(2A)-adrenergic deficient mice. The decreases in arterial pressure were nearly identical to those of wild type mice, whereas bradycardia was attenuated. Thus, in the mouse moxonidine acts within the RVLM region to lower arterial pressure mainly through the I(1)-imidazoline receptor independent of alpha(2)-adrenergic receptors. PMID- 10799666 TI - Retinas from albino rats are more susceptible to ischaemic damage than age matched pigmented animals. AB - Age- and sex-matched pigmented (Lister Hooded) and albino (Wistar) rats were used in this study. The retinas of the animals were subjected to pressure-induced ischaemia (35 min, 120 mmHg) and reperfusion (3 days) in precisely the same way. The b-wave of the electroretinogram (ERG) in the pigmented animals recovered to normal levels while those of the albino rats were reduced by more than 80%. Moreover, the choline acetyltransferase (ChAT) immunoreactivity associated with a sub-set of amacrine cells was almost completely obliterated in the retinas from the albino rats but unaffected in the retinas of the pigmented rats. Also, in certain areas of the retina from albino rats there was a suggestion that the calretinin-immunoreactivity was affected. This was never seen in the retinas of the pigmented animals. The GABA-immunoreactivity in the retina of both albino and pigmented rats appeared to be unaffected by ischaemia/reperfusion. The data presented show that retinas from albino rats are more susceptible to ischaemia/reperfusion than retinas from pigmented animals. The results also show that reduction of the b-wave of the ERG and changes in the nature of the ChAT immunoreactivity represent sensitive markers to detect the effect of ischaemia/reperfusion to the retina. PMID- 10799667 TI - Ovarian cycle-related changes of glial fibrillary acidic protein (GFAP) immunoreactivity in the rat interpeduncular nucleus. AB - The interpeduncular nucleus (IPN) of female rats was studied across the estrous cycle to observe whether the expression of the astroglial marker, glial fibrillary acidic protein (GFAP) reacts to hormonal changes in an area not belonging to the 'endocrine brain'. A marked reduction of immunoreactive GFAP was observed in estrus as compared to the immunoreactivities in met- and proestrus. This finding is consistent with earlier observations in the endocrine hypothalamus, but also proves that gonadal steroids influence astroglia in brain regions not involved in neuroendocrine regulation. Since cyclic fluctuations of synaptic numbers in the female have been described only for the endocrine hypothalamus, decrease of immunoreactive GFAP in the IPN during estrus may reflect a down-regulation of GFAP synthesis. PMID- 10799668 TI - Effects of cocaine context on NAcc dopamine and behavioral activity after repeated intravenous cocaine administration. AB - In two conditioning experiments, identical procedures (previously shown to produce place preferences for a cocaine-paired environment) were used to assess dopaminergic and behavioral activity correlates of cocaine reward conditioning and sensitization. In these experiments, animals received repeated injections of intravenous cocaine (4.2 mg/kgx6) or saline (0.2 mlx6) on alternating days. One group in each of these experiments ('Cocaine Cues') occupied a consistent distinctive environment during cocaine treatments and testing sessions. For the other conditioned group ('Novel'), all procedures were the same, except that the last cocaine injection was administered while animals were occupying a novel environment. During day 1 and day 6 of the cocaine treatment, behavioral activity was assessed in experiment 1 and in vivo microdialysis procedures were conducted in experiment 2. Over the course of the conditioning sessions, cocaine-induced behavioral activity (locomotion and rearing) increased significantly in the Cocaine Cues group, but not in the Novel group. In addition, cocaine-induced increases in NAcc dopamine levels were significantly greater when cocaine experienced animals were tested in a cocaine-paired environment compared to equally experienced and cocaine-naive animals tested in a novel environment. Context-dependent behavioral sensitization is a well-documented phenomenon. The observation of a corresponding enhancement of dopamine efflux in lieu of a lengthy withdrawal period is uncommon, but can be attributed to methodological differences across studies. The present study uniquely demonstrates concurrent context-dependent potentiation of behavioral and dopaminergic responses to cocaine occurring in conjunction with cocaine reward. PMID- 10799669 TI - Clomethiazole is neuroprotective in models of global and focal cerebral ischemia when infused at doses producing clinically relevant plasma concentrations. AB - We investigated the neuroprotective effect of infusing various doses of clomethiazole in models of global and focal cerebral ischemia. In a model of global ischemia, gerbils were infused with clomethiazole (intravenous), attaining steady state plasma concentrations of between 1 and 13 microM for 24 h. In a transient middle cerebral artery occlusion model in rats, clomethiazole was administered subcutaneously over 22.75 h using osmotic minipumps producing steady state plasma concentrations of between 1 and 13 microM. Clomethiazole was protective in these models at plasma concentrations of respectively 6.1 microM and above and 3.5 microM and above. Clomethiazole is thus neuroprotective in both global and focal ischemia at plasma concentrations known to be well tolerated in stroke patients. PMID- 10799670 TI - Early microglial reaction following mild forebrain ischemia induced by common carotid artery occlusion in rats. AB - Early microglial reaction following mild ischemic injury caused by bilateral common carotid artery occlusion has been investigated in rats. The ischemic insults lasted for 10, 15 and 20 min without recirculation, and with several reperfusion intervals from 1 h to 3 days. The resting and activated microglial cells were visualized with immunohistochemistry using monoclonal antibodies raised against the CR3 complement receptor, the MHC class I and class II antigens, the macrophage common antigen and with Bandeiiraea simplicifolia lectin histochemistry. The neuroprotective effect of hypothermia on the early microglial activation was also studied. Ten minutes bilateral common carotid artery occlusion in hypothermic rats without reperfusion caused a mild microglial reaction in the hippocampus. Strong reaction was seen following 20 min insult without reperfusion. Ischemia followed by recirculation caused milder reaction than without reperfusion. Our results suggest that the microglial cells are very sensitive indicators of a mild, transient ischemic insult that does not result in neuronal cell death. PMID- 10799671 TI - Physiology and pharmacology of native glycine receptors in developing rat ventral tegmental area neurons. AB - Properties of whole-cell glycine-activated currents (I(Gly)) of neurons freshly isolated from ventral tegmental area of rats between postnatal day (P) 0 and 40 (P0-P40) were examined using the gramicidin-perforated patch technique. I(Gly) was present throughout the period of investigation and displayed age-related alternations in its kinetics, reversal potentials (E(Gly)) and sensitivity to antagonists. During development, the time constant for desensitization of I(Gly) decreased about two-fold, and the E(Gly) shifted to more negative potentials. Specifically, E(Gly) was -29 mV in neonatal (P0-P7) and decreased to -50 mV in adult (P24-P40) neurons. While the neonatal glycine receptors (GlyRs) are resistant to strychnine, the adult GlyRs are sensitive to STR. The values for 50% inhibition (IC(50)) of I(Gly) was 110 and 12 nM in P0-P3 and P24-P40 neurons, respectively. Picrotoxin, on the other hand, was more effective in blocking neonatal neurons. The IC(50) values were 17 and 231 microM for the neonatal and adult neurons, respectively. These observations are discussed in terms of developmental changes of subunit composition of GlyRs and of intracellular Cl(-) concentrations. PMID- 10799672 TI - Intracellular glutathione levels determine cerebellar granule neuron sensitivity to excitotoxic injury by kainic acid. AB - Glutathione (GSH) is a key component of the cellular defence cascade against injury caused by reactive oxygen species. Kainic acid (KA) is a potent central nervous system excitotoxin. KA-elicited neuronal death may result from the generation of ROS. The present study was undertaken to characterize the role of GSH in KA-induced neurotoxicity. Cultures of cerebellar granule neurons were prepared from 8-day-old rats, and used at 8, 14 and 20 days in vitro (DIV). Granule neurons displayed a developmental increase in their sensitivity to KA injury, as quantified by an ELISA-based assay with the tetrazolium salt MTT. At DIV 14 and 20, a 30-min challenge with KA (500 microM) reduced cell viability by 45% after 24 h, significantly greater (P<0.01) than the 22% cell loss with DIV 8 cultures. Moreover acute (30 min) KA exposure concentration-dependently reduced intracellular GSH and enhanced reactive oxygen species generation (evaluated by 2', 7'-dichlorofluorescein diacetate). In comparison to control, KA (500 microM) lowered GSH levels in DIV 8 granule neurons by 16% (P=0. 0388), and by 36% (P=0.0001) in both DIV 14 and DIV 20 neurons, after 30 min. Preincubation of granule neurons with the membrane permeant GSH delivery agent, GSH ethyl ester (5 mM), for 30 min significantly increased intracellular GSH content. Importantly, GSH ethyl ester reduced the toxic effects of KA, becoming significant at 1 mM (P=0.007 vs. KA-treated group), and was maximal at >/=2.5 mM (P<0.0001). GSH ethyl ester displayed a similar dose-dependence in its ability to counteract KA induced depletion of cellular GSH. The data strengthen the notion that cellular GSH levels have a fundamental role in KA-induced neurotoxicity. PMID- 10799674 TI - The pain inhibiting pain effect: an electrophysiological study in humans. AB - This study examines the counterirritation phenomenon of experimental pain in human subjects. Phasic pain induced by intracutaneous electrical stimuli was simultaneously applied with tonic pain induced by ischemic muscle work. Pain ratings, spontaneous EEG and evoked potentials were measured. We found a significant reduction of phasic pain ratings during and 10 min after tonic pain. The late somatosensory evoked potentials as neurophysiological correlates of phasic pain sensation were attenuated until 20 min after tonic pain offset. The extent of phasic pain relief due to concomitant tonic pain was small but significant, comparable to the effect of a regular systemic dose of a narco analgesic drug in this experimental pain model. On the other hand, no modulations in the late components of the auditory evoked potential and the power spectrum of the spontaneous EEG were observed. These variables reflect the attention and vigilance of the subject and are well-known to be affected by opioids. The only exception was an increase of beta power, which might reflect hyperarousal during tonic pain. These results support the suggestion, that the analgesic effect of heterotopic noxious stimulation in humans is based on the activation of a specific inhibitory pain control system. Systemic release of endogenous opioids is unlikely to be involved, because the typical effects of opioids on the EEG were not observed. PMID- 10799673 TI - Neuronal activity in female rat preoptic area associated with sexually motivated behavior. AB - Single unit activities were recorded from 31 neurons in the preoptic area (POA) of female rats engaging in sexual interactions. Concurrent videotape recordings were used to establish a relationship between neuronal activity and particular behavioral events. In 14 of the 31 neurons, the firing rate changed in association with bouts of sexual activity. The remaining 17 fired with more variability regardless of episodes of sexual interactions. Peri-event histograms identified four types of neurons: type 1 (n=4) increased their firing rate when the female rats initiated proceptive behavior; type 2 (n=4) showed a brief activation when the male mounted; type 3 (n=4) fired in response to intromission, and type 4 (n=2) were inhibited prior to and throughout the display of lordosis reflex. Type 1 neurons fired at significantly higher rates during the solicitatory period, from the initiation of solicitatory locomotion to the male mounts. Their activity was suppressed when the males mounted successfully with intromission. Types 1-3 neurons were recorded from the transitional region between the medial and lateral POAs. Type 4 neurons were located more medially in the medial POA. Systemic injection of pimozide, a dopamine receptor blocker, diminished firing in type 1 neurons and abolished proceptivity. The firing pattern in type 1 neurons appeared to embody the motivational state of the animal with an implication for a consummatory value of penile intromission. Visceral or somatosensory inputs may be responsible for short bursts in types 2 and 3 neurons. Type 4 neurons behaved exactly as if they inhibit the execution of the lordosis reflex. The results showed separate sets of POA neurons each specifically associated with proceptive and receptive components of female rat sexual behavior. PMID- 10799675 TI - MRZ 2/579, a novel uncompetitive N-methyl-D-aspartate antagonist, reduces infarct volume and brain swelling and improves neurological deficit after focal cerebral ischemia in rats. AB - The purpose of this study was to evaluate the effects of MRZ 2/579, an uncompetitive N-methyl-D-aspartate antagonist, on infarct size, extent of swelling and neurological deficit in a model of transient middle cerebral artery occlusion in rats. Physiologically controlled Sprague-Dawley rats received 2 h MCAo by retrograde insertion of an intraluminal suture coated with poly-L-lysine. The agent (MRZ 2/579) or vehicle (sodium chloride 0.9%) was administered i.v. immediately after suture removal following a 2-h period of MCAo. Two experimental groups were studied: group A was treated by vehicle (bolus infusion:1 ml/kg for 10 min followed by infusion of 6 ml/kg/h over 6 h). Group B was treated by MRZ 2/579 (bolus infusion:10 mg/kg for 10 min followed by infusion of 6 mg/kg/h over 6 h). The neurological status was evaluated during occlusion (at 60 min) and daily for 3 days after MCAo. Brains were then perfusion-fixed, and infarct volumes and brain swelling were determined. MRZ 2/579 significantly improved the neurological score compared to vehicle-treated rats at 48 h (6.2+/-0.6 and 8.7+/ 0.5, respectively; P<0.004) and 72 h after MCAo (5.2+/-0.6 and 8.4+/-0.5, respectively; P<0.001). Treatment with MRZ 2/579 also significantly reduced total infarct volume (29.3+/-11.1 and 83.2+/-16.5 mm(3), respectively; P<0. 01), cortical infarct volume (24.8+/-11.2 and 70.0+/-18.0 mm(3), respectively; P<0.04) and subcortical infarction (21.2+/-4.1 and 49. 6+/-4.5 mm(3), respectively; P<0.0002). Brain swelling was also markedly reduced compared with vehicle-treated rats (4.7+/-1.3 and 10.8+/-2.1%, respectively; P<0.02). These results demonstrate that treatment with MRZ 2/579, when administered promptly after reperfusion, confers neuroprotective effects on infarct volume, brain swelling, and neurological score compared to the vehicle group. PMID- 10799676 TI - Effects of differential modulation of mu-, delta- and kappa-opioid systems on bicuculline-induced convulsions in the mouse. AB - The present study investigated the effects of micro-, delta- and kappa-opioid receptor agonists on seizures produced by blockade of gamma-aminobutyric acid (GABA)-mediated synaptic transmission in the mouse. The selective GABA(A) receptor antagonist bicuculline (1.25-3 mg/kg) given subcutaneously caused dose dependent clonic-tonic convulsions. These convulsions were potentiated by the prototypic mu-opioid receptor agonist morphine given subcutaneously 20 min prior to a subconvulsive dose of bicuculline. The potentiation by morphine was completely reversed by pretreatment intraventricularly with the selective mu opioid receptor antagonist beta-funaltrexamine (0.5 microgram/mouse). Pretreatment intraventricularly with the selective delta-opioid receptor agonists 2-methyl-4aalpha-(3-hydroxyphenyl)-1,2,3,4,4a,5,12, 12abeta-octahydro quinolino[2,3,3-g]isoquinoline ((-)TAN-67) or [D-Pen(2,5)]-enkephalin (DPDPE) showed a dose-dependent increase in the incidence of convulsions. Pretreatment with naltrindole (2 mg/kg, s.c.), a selective delta-opioid receptor antagonist, abolished the enhancement of the bicuculline-induced convulsions by DPDPE. In contrast, pretreatment with the selective kappa-opioid receptor agonist U-50,488H (0.6-80 mg/kg, subcutaneously or 25-100 microgram/mouse, intraventricularly) produced a dose-dependent suppression of the bicuculline-induced convulsions. The inhibitory effect of U-50,488H was completely blocked by pretreatment subcutaneously with nor-binaltorphimine (5 mg/kg), a selective kappa-opioid receptor antagonist. This study demonstrates that activation of both mu- and delta-opioid receptors increases the incidence of convulsions produced by blockade of GABA-mediated synaptic transmission, while stimulation of kappa opioid receptors has an anticonvulsive effect. PMID- 10799677 TI - The role of GABAergic inhibition on direction-dependent sharpening of frequency tuning in bat inferior collicular neurons. AB - This study examined the role of GABAergic inhibition on direction-dependent sharpening of frequency tuning curves (FTCs) in bat inferior collicular (IC) neurons under free field stimulation conditions. The minimum threshold (MT) at the neurons best frequency (BF) and the sharpness (Q(10), Q(20), Q(30)) of FTCs of most IC neurons increased as the sound direction changed from contralateral azimuths to ipsilateral azimuths. The application of GABA(A) antagonist, bicuculline, lowered all MTs but the application did not abolish direction dependent variation in MT. MTs determined during bicuculline application at 40 ipsilateral were still significantly higher than those determined at 40 degrees contralateral (two-tailed paired t-test, P<0.0001). In contrast, although application of bicuculline essentially had no effect on the BFs of IC neurons, it differentially broadened neurons FTCs at different azimuths abolishing the direction-dependent sharpening of frequency tuning (i. e. Q(n) values, two-tailed paired t-test, P<0.01). These data indicate that GABAergic inhibition makes an important contribution to the direction-dependent frequency tuning of most IC neurons. PMID- 10799678 TI - Conditioned c-Fos in mouse NTS during expression of a learned taste aversion depends on contextual cues. AB - c-Fos expression in the nucleus tractus solitarius (NTS) of the rat has been found to follow administration of a variety of pharmacologically diverse unconditioned stimuli (US), and it has been proposed that NTS is a critical structure in transduction of the US during taste aversion learning. Before conditioning, the conditioned stimulus (CS) taste does not induce c-Fos in NTS, but following pairing of the CS and US, subsequent CS presentation induces c-Fos in NTS. Although it has been suggested that the shift in the c-Fos response following conditioning represents a molecular correlate of taste aversion learning, i.e. the formerly neutral CS now predicts the toxicity associated with the US, the data presented here suggest a more cautious interpretation of c-Fos expression in NTS. In mice, post-conditioning c-Fos expression to the CS depends on contextual cues: when conditioning and testing occur in a novel environment, CS saccharin causes an increase in c-Fos expression, and when conditioning and testing occur in the home cage, CS saccharin produces a decrease in c-Fos expression relative to controls. Furthermore, we show that merely placing an animal into a novel environment is sufficient to drive c-Fos expression in NTS. These data suggest that c-Fos expression in NTS can be driven by a number of different stimuli and conditions, and that these responses may depend on context dependent activation of forebrain structures shown to drive conditioned c-Fos expression in NTS. PMID- 10799679 TI - Central 5-HT(1) and 5-HT(2) binding sites in transgenic mice with reduced glucocorticoid receptor number. AB - Transgenic mice bearing a transgene coding for a glucocorticoid receptor antisense mRNA, which partially blocks glucocorticoid receptor expression, were used in order to clarify the role of glucocorticoid receptors in the regulation of 5-HT(1A), 5-HT(1nonA) and 5-HT(2) binding sites labelled by quantitative autoradiography in the frontal and prefrontal cortex, striatum, hypothalamus, amygdala and raphe nuclei. We found that 1 nM [3H]8-hydroxy-2-[di-N propylamino]tetralin ([3H]8-OH-DPAT) binding to 5-HT(1A) sites was decreased in strata oriens (-15.1+/-3.5%) and radiatum-lacunosum-moleculare (-13.3+/-4.3%) of the hippocampal CA(3) area, and 2 nM [3H]5-hydroxytryptamine binding to 5 HT(1nonA) sites in the presence of 100 nM 8-OH-DPAT and mesulergine was decreased in the dorsal subiculum (-17.8+/-6.9%). By contrast, 5-HT(2) sites labelled by 0.5 nM of (+/-)-1-(2, 5-dimethoxy-4-[125I]iodophenyl)-2-aminopropane was increased in the dorsal subiculum (+35.2+/-11.5%) and CA(2) area (+29.2+/-11.3%). The observed differences in binding to 5-HT(1) and 5-HT(2) sites were all located in areas of the hippocampus that contain both gluco- and mineralo-corticoid receptors, and no difference was observed in anatomical structures which contain only glucocorticoid receptors. Therefore, it seems that the important factor for the regulation of these 5-HT receptors is the interaction between gluco- and mineralo-corticoid receptors rather than the absolute density of glucocorticoid receptors. These results suggest that some of the alterations of the serotonergic neurotransmission observed in depressed patients might be secondary to an altered glucocorticoid receptor function. PMID- 10799680 TI - DARPP-32 and CREB are present in type 2 iodothyronine deiodinase-producing tanycytes: implications for the regulation of type 2 deiodinase activity. AB - Type 2 iodothyronine deiodinase, an enzyme involved in the conversion of thyroxin to the biologically active 3,5, 3'-triiodothyronine, is highly concentrated in a group of specialized ependymal cells, tanycytes, lining the wall and floor of the third ventricle. As this distribution is highly reminiscent of the distribution of cells containing the phosphatase inhibitor, DARPP-32, we raised the possibility that these two proteins may coexist in tanycytes and that DARPP-32 may modulate type 2 deiodinase activity by regulating the phosphorylation state of the cAMP regulatory factor, CREB. To address this question, double-labeling histochemical studies were performed for type 2 deiodinase mRNA and DARPP-32 immunoreactivity (IR), or DARPP-32- and CREB-IR in the same tissue sections. Type 2 deiodinase mRNA was found in the cell bodies of all DARPP-32-immunolabeled tanycytes. Both type 2 deiodinase mRNA and DARPP-32-IR also extended into tanycyte processes that ramified in the arcuate nucleus and median eminence, in close association with blood vessels and portal capillaries. In contrast, type 2 deiodinase mRNA was not present in the same cells that contained DARPP-32-IR in the pituitary gland. All tanycytes containing DARPP-32-IR also contained CREB-IR in their nucleus. Since type 2 deiodinase activity can be induced by substances that increase cAMP, we hypothesize that DARPP-32 may regulate the activity of type 2 deiodinase by prolonging the activation of CREB. Selectivity for the colocalization of these factors to tanycytes but not the pituitary gland, may explain the heterogeneous response of type 2 deiodinase activity in these two loci in response to specific stimuli such as fasting. PMID- 10799681 TI - HPLC and electrospray ionization mass spectrometry as tools for the identification of APGWamide-related peptides in gastropod and bivalve mollusks: comparative activities on Mytilus muscles. AB - The APGWamide-related neuropeptides, predicted by the cDNA of the APGWamide precursor of Mytilus edulis, have been sought by means of HPLC and electrospray mass ionization. The three predicted peptides KPGWamide, RPGWamide and TPGWamide were detected in the three main muscles and surprisingly an ion at m/z 429 corresponding to the gastropod peptide APGWamide was also demonstrated. Similar investigations performed in Lymnaea stagnalis central nervous system (CNS) revealed the occurrence of mussel APGWamide-related peptides (APGWamide-RPs) demonstrating for the first time the presence and the expression of the two precursors in both gastropod and bivalve mollusks. The absence of homologous domain in the Mytilus precursor [P. Favrel, M. Mathieu, Molecular cloning of a cDNA encoding the precursor of Ala-Pro-Gly-Trp-amide related neuropeptides from the bivalve Mytilus edulis. Neurosci. Lett. 1996;205:210-214] and the Lymnaea precursor [A.B. Smit, C.R. Jimenez, R.W. Dirks, R.P. Croll, W.P.M. Geraerts, Characterization of cDNA clone encoding multiple copies of the neuropeptide APGWamide in the molluscs Lymnaea stagnalis. J. Neurosci. 1992;12:1709-1715] eliminates the hypothesis of an alternative splicing of a single gene and suggests the likelihood of two genes probably resulting from duplication of an ancestral gene before the divergence between gastropods and bivalves. The similar potency observed on contraction assay and the differential distribution of the various peptides suggest that they may exert distinct activities on multiple targets. PMID- 10799682 TI - GABAergic neurons in the rabbit visual cortex: percentage, layer distribution and cortical projections. AB - 6250 neurons yielding either callosal or inter-areal ipsilateral projections extrinsic to area 17 was GABAergic. Comparing these findings with those reported for other mammals, it seems that the incidence and distribution of GABAergic neurons in the visual cortex is similar in rabbits and rats. In contrast to rats but akin to higher mammals, no GABAergic neuron was found to furnish cortico cortical connections to area 17 other than intrinsic connections. PMID- 10799683 TI - Docosahexaenoic acid (DHA) improves the age-related impairment of the coupling mechanism between neuronal activation and functional cerebral blood flow response: a PET study in conscious monkeys. AB - The regional cerebral blood flow (rCBF) response to vibrotactile stimulation was compared in conscious young (5.9+/-1.8 years old) and aged (18.0+/-3.3 years old) monkeys using [15O]H(2)O and high-resolution positron emission tomography (PET). In addition, the effects of docosahexaenoic acid (DAH), an n-3 polyunsaturated fatty acid (PUFA), on the rCBF response to stimulation were evaluated in aged monkeys. Soybean milk (SBM) or DHA-containing SBM (DHA/SBM) was supplied to aged monkeys for 1 and 4 weeks. Under control conditions, vibrotactile stimulation elicited an increase in the rCBF response in the contralateral somatosensory cortices of both young and aged monkeys, but the degree of increase in the rCBF response was significantly lower in aged monkeys (116% of corresponding 'rest' condition) than that in young animals (141%). The regional cerebral metabolic rate of glucose (rCMRglc) response to the stimulation, an index of neuronal activation, was not significantly different between young and aged monkeys as measured by [18F]-2-fluoro-2-deoxy-D-glucose (FDG). Supply of DHA/SBM at a dose of 150 mg/kg/day as DHA for 1 week resulted in a significant increase in rCBF response to stimulation (127%) in aged monkeys, and 4-week supply of DHA induced further facilitation of the rCBF response (133%). In contrast, the reduced rCBF response in the aged monkeys was not affected by SBM alone for either 1 or 4 weeks. The neuronal activation itself elicited by the stimulation, as measured by [18F]FDG, was not affected by SBM or DHA/SBM. These results suggested the involvement of DHA in the coupling mechanism between the neuronal activation and rCBF response, possibly by modulation of cholinergic neuronal transmission. PMID- 10799684 TI - A function of delayed rectifier potassium channels in glial cells: maintenance of an auxiliary membrane potential under pathological conditions. AB - Muller glial cells from human and guinea-pig retinae were investigated using the whole-cell patch-clamp technique. Human Muller cells from eyes with different diseases were characterized by diminished inwardly-rectifying K(+) currents. A comparable reduction of these currents was achieved in guinea pig Muller cells by treatment with iodoacetate to generate ischemia-like conditions. Consequently, the membrane potentials were reduced significantly in both diseased human and iodoacetate-treated guinea-pig Muller cells as compared to normal controls. However, the potentials were still clearly negative. Delayed rectifier currents could still be recorded under these conditions. Application of quinine blocked the delayed rectifier K(+) channels, and resulted in a total breakdown of the membrane potentials. Thus, it becomes apparent that the glial delayed rectifier K(+) channels are necessary to maintain an 'auxiliary' membrane potential under certain pathological conditions that are characterized by an almost total loss of inward rectifier conductance. Therefore, the delayed rectifier K(+) channels of glial cells may become crucial for the support of basic glial functions. PMID- 10799685 TI - Changes in breathing during observation of effortful actions. AB - Respiration and heart rates were recorded in normal subjects watching effortful actions produced by an actor in front of them. Subjects remained immobile throughout. Two experiments were performed. In experiment 1, subjects watched a weight-lifting performance, either static or dynamic, with increasing weights. In experiment 2, they watched a walking/running performance on a treadmill moving at increasing speed. In both experiments, no change was found in observers' heart rate. By contrast, consistent changes were found in respiration rate. These changes tended to follow the exercise rhythm of the actor, specially during accelerated running (from 2.5 to 10 km/h) where respiration rate increased linearly with speed of the treadmill. Average maximum increase ranged between 25 and 30% above resting rate. This finding demonstrates activation of central mechanisms related to action performance during observation of effortful actions. It could represent a basis for understanding and imitating actions performed by other people. PMID- 10799686 TI - Calcium transient activity in cultured murine neural crest cells is regulated at the IP(3) receptor. AB - In a previous study we have shown that cultured neural crest cells exhibit spontaneous calcium transients and that these events are required for neurogenesis. In this study, we examine the mechanism that generates these calcium transients. Extracellular Ca(2+) modulates calcium transient activity. Lanthanum (La(3+)), a general calcium channel antagonist and zero extracellular Ca(2+), reduces the percentage of cells exhibiting calcium transients (26.2 and 40. 5%, respectively) and decreases calcium spiking frequency (4.5 to 1. 0 and 2.5 to 1.0 spikes/30 min, respectively). Intracellular calcium stores also contribute to the generation of calcium transients. Depleting the calcium stores of the endoplasmic reticulum (ER) reduces the percentage of active cells (15.7%) and calcium spiking frequency (2.8 to 1.5 spikes/30 min). Ryanodine (100 microM), which blocks calcium release regulated by the ryanodine receptor (RyR), had no effect on calcium transient activity. Blocking inositol 1,4, 5-triphosphate receptor (IP(3)R)-dependent calcium release, with elevated extracellular Mg(2+) (20 mM), abolished calcium transient activity. Mg(2+) did not block caffeine sensitive calcium release (RyR-dependent) or voltage dependent calcium channels. Mg(2+) also suppressed thimerosal-induced calcium oscillations (IP(3)R dependent). Small increases in the intracellular calcium concentration ([Ca(2+)](i)), increases the percentage of active cells and the calcium spiking frequency, while larger increases in [Ca(2+)](i) block the transients. Reducing intracellular IP(3) levels reduces the percentage of active cells and the calcium spiking frequency. We conclude that the mechanism for generating spontaneous calcium transients in cultured neural crest cells fits the model for IP(3)R dependent calcium excitability of the ER. PMID- 10799687 TI - Bidirectional actions of docosahexaenoic acid on hippocampal neurotransmissions in vivo. AB - Docosahexaenoic acid (DHA), a 22-carbon fatty acid with six double bonds, is one of the major polyunsaturated fatty acids in fish oils or in the mammalian central nervous system and is believed to be essential for neuronal plasticity and development. In the present study, we evaluated the effect of DHA on hippocampal neurotransmissions using anesthetized rats. Field excitatory postsynaptic potential (fEPSP) evoked by stimulation of the Schaffer collaterals was recorded from the CA1 stratum radiatum. Following intracerebroventricular injection of DHA 25 nmol, the fEPSP slope decreased gradually in 30 min and was eventually suppressed by about 30%. On the other hand, when fEPSP was evoked by stimulation of the perforant path was recorded in the molecular layer of the dentate gyrus, an increase in fEPSP slope occurred over a similar time course after DHA injection. These phenomena were independent of N-methyl-D-aspartate receptor activity. Linoleic acid, one of polyunsaturated fatty acids, was virtually ineffective. Furthermore, we investigated the effect of DHA on hippocampal synaptic plasticity. Although DHA did not alter the profile of paired-pulse facilitation, it inhibited the induction of long-term potentiation in the CA1 area but not in the dentate gyrus. Thus, DHA exerts regionally different effects on hippocampal neurotransmission and may be a good tool for clarifying physiological functions of the hippocampus. PMID- 10799688 TI - The role of pH and osmolarity in evoking the acetic acid-induced wiping response in a model of nociception in frogs. AB - Acetic acid applied to the hindlimb of a frog evokes a vigorous wiping of the exposed skin. The aim of this study was to determine if acetic acid evokes this wiping response by decreasing subepidermal pH. Because acetic acid is hyperosmolar, a second aim was to determine if the osmolarity of acetic acid contributed to evoking the wiping response. In behavioral experiments, different acids or acetic acid/sodium acetate buffers at different pHs were used to evoke the wiping response. In separate experiments, subepidermal pH was measured in vitro while these same solutions were applied to samples of skin from frogs. The wiping response evoked by acetic acid was associated with a decrease in subepidermal pH to a level that has been shown to activate nociceptors. Interestingly, formic, oxalic, sulfuric, and hydrochloric acid evoked the wiping response without decreasing subepidermal pH. The osmolarity of acetic acid contributed to evoking the wiping response because buffers at subthreshold pHs evoked the wiping response. Also, the osmolarity required to evoke the wiping response depended upon the pH of the buffer. Thus, acetic acid and the buffers at pH 2.97 and 4.67 could evoke the wiping response by decreasing subepidermal pH. In contrast, formic, oxalic, sulfuric, and hydrochloric acid, as well as the buffers at pH 5.17 and 5.67, evoked the wiping response through another mechanism, perhaps by increasing subepidermal osmolarity. These studies demonstrate that both pH and osmolarity may contribute to nociception produced by algesic chemicals and may be important in inflammatory pain. PMID- 10799689 TI - Modulation of G-protein linked cAMP accumulation in immortalized murine cortical astrocytes by retroviral infection. AB - The present study characterized beta-adrenergic receptors (beta-AR) in a clonal cell line (C1) immortalized from cerebral cortical astroglial cells of FVB/N mice. We also determined whether the wild type Moloney murine leukemia virus (wt MoMuLV) and one of its neuropathogenic mutants, ts1-MoMuLV, modulated the beta-AR system in these cells. We observed that C1 cells possess a functional beta-AR system coupled to cAMP accumulation and capable of normal agonist-induced regulation (desensitization). Significant increases were observed in forskolin stimulated cAMP accumulation in C1 cells infected by wt MoMuLV and by ts1-MoMuLV. In contrast, the cAMP response to beta-AR stimulated by isoproterenol was relatively spared after viral exposure. PMID- 10799690 TI - Striatal oxidative damage parallels the expression of a neurological phenotype in mice transgenic for the mutation of Huntington's disease. AB - We examined the degree of oxidative damage to the brain of mice transgenic for the mutation responsible for Huntington's disease. We found that there is a progressive increase in striatal lipid peroxidation (LP), that parallels the worsening of the neurological phenotype. We consider that these transgenic mice may provide an interesting system to test treatments aimed at protecting cells from damage induced by free radicals. PMID- 10799691 TI - Effect of N-methyl-D,L-aspartate (NMA) on gonadotropin-releasing hormone (GnRH) gene expression in male mice. AB - The glutamate analog N-methyl-D,L-aspartate (NMA) affects the regulation of GnRH and LH release in mammals. Several laboratories have reported a rapid and transient increase in GnRH mRNA levels of male rats after NMA injection. Studies employing the simultaneous measurements of nuclear GnRH primary transcript RNA, a reflection of gene transcription, and GnRH mRNA suggest that NMA's effect on GnRH gene expression in the rat is likely due to post-transcriptional regulation. Despite the increasingly widespread use of transgenic mice, surprisingly little is known about the regulation of GnRH gene expression in the mouse. In this study, we assessed in detail the effects of NMA on GnRH gene expression in adult male mice. In the first experiment, GnRH mRNA levels in mice killed 60-min post NMA injection (20 mg/kg bw, ip; n=9/treatment group) were lower (P<0.05) when compared to controls (saline vehicle). In the second experiment, mice (n=7/treatment group) were administered NMA or saline vehicle and were killed at 15-, 60- and 120-min post-injection. Consistent with the first experiment, treatment with NMA resulted in a significant decrease (P<0.05) in cytoplasmic GnRH mRNA compared to control levels at 15- and 60-min but not 120-min. NMA treatment decreased the nuclear GnRH primary transcript RNA at 120-min but not at earlier time points. In summary, we have shown that regulation by NMA of GnRH gene expression in mice differs substantially from rats. This differential regulation of GnRH gene expression between rats and mice warrants further investigation. PMID- 10799692 TI - The 5-HT(1A) antagonist WAY 100635 can block the low-dose locomotor stimulant effects of cocaine. AB - Cocaine treatments of 2.5, 5.0, 10.0 and 15.0 mg/kg induced dose-dependent increases in locomotor behavior. This cocaine-induced increase in locomotion was blocked if the animals were administered the selective 5-HT(1A) antagonist, WAY 100635 (0.4 mg/kg) prior to the cocaine treatment. The 0.4-mg/kg dose of WAY 100635 did not affect locomotor behavior or alter cocaine availability in brain. PMID- 10799693 TI - Selenium deficiency potentiates methamphetamine-induced nigral neuronal loss; comparison with MPTP model. AB - The present study was designed to understand the role of an antioxidant, selenium (Se) on methamphetamine (MA)-induced dopaminergic cell damage in the substantia nigra (SN). Male C57BL/6J mice were fed either selenium-deficient (<0.01 ppm Se) or selenium-replete (0.2 ppm Se) diet for 90 days. Se-deficiency potentiates MA induced reductions of tyrosine hydroxylase-like immunoreactivity (TH-IR), dopamine (DA) and its metabolites, 3, 4-dihydroxyphenylacetic acid (DOPAC) and homovanilic acid (HVA) in the SN. These dopaminergic toxicities were comparable to that induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). By contrast, Se-repletion significantly blocked dopaminergic toxicity after MA treatments. These results suggest that Se-deficient MA-treated mouse is a relevant model of Parkinsonism, and that optimal level of Se plays a crucial role in preventing nigral dopaminergic toxicity induced by MA. However, different mechanisms in the thermoregulation mediated by MA or MPTP remain to be further determined. PMID- 10799694 TI - Cyclin-dependent kinase 5 (Cdk5) associated with Lewy bodies in diffuse Lewy body disease. AB - We investigated the cyclin-dependent kinase (Cdk) 5 distribution pattern in diffuse Lewy body disease brains using immunohistochemistry. Cdk5 immunoreactivity was detected in both brainstem-type Lewy bodies (LBs) and cortical LBs. The number of Cdk5-positive LBs was less than that of ubiquitin- or alpha-synuclein-positive LBs, and more than that of phosphorylated neurofilament positive LBs. Immunoelectron microscopy revealed Cdk5-immunolabeled granulo filamentous components in LBs and LB-related neurites. These data suggest that Cdk5 may be associated with LB formation. PMID- 10799695 TI - Modulation of the formalin-induced nociceptive response by diabetes: possible involvement of intracellular calcium. AB - We examined the involvement of cytosolic calcium in the modulation of the formalin-induced nociceptive response by diabetes. Injection of formalin into the hindpaw of mice produced a biphasic nociceptive response consisting of immediate (first phase) and tonic (second phase) components. Although the duration of the first-phase response was significantly longer in diabetic mice than in non diabetic mice, the second phase was significantly shorter in diabetic mice. The first-phase response was dose-dependently and significantly reduced by pretreatment with ryanodine, which blocks Ca(2+) release from Ca(2+)/caffeine sensitive microsomal pools. The second-phase response was also significantly increased when diabetic mice were pretreated with ryanodine. However, ryanodine had no significant effect on either the first-phase or second-phase response in non-diabetic mice. On the other hand, pretreatment with thapsigargin, which inhibits Ca(2+) uptake into the inositol-1,4, 5-trisphosphate-sensitive microsomal Ca(2+) pool, significantly enhanced the first-phase response in non diabetic mice. Furthermore, thapsigargin significantly and dose-dependently reduced the second phase of the formalin-induced nociceptive response in non diabetic mice. Thapsigargin administered i.t. did not significantly affect either the first- or the second-phase response in diabetic mice. These results suggest that the change in the formalin-induced nociceptive response in diabetic mice may be due, at least in part, to the modification of nociceptive transmission in the spinal cord by intracellular calcium. PMID- 10799696 TI - Attenuation of free radical generation during reversible focal cerebral ischemia with the nitric oxide inhibitor, L-NAME (L-N(G)-nitro-L-arginine methyl ester). AB - The role of oxygen free radical generation during reversible focal cerebral ischemia and its relationship to nitric oxide mediated mechanisms were examined. In this study, a left frontal cortex microdialysis probe was placed into the previously defined ischemic penumbra region and perfused with a salicylate/CSF solution in the presence or absence of the nitric oxide synthase (NOS) inhibitor L-NAME. Rats were then subjected to transient left hemisphere focal cerebral ischemia. Dialysate was collected at baseline and during the ischemic/reperfusion phase, and the hydroxylation products of salicylate were measured by HPLC with electrochemical detection. A significant elevation of free radical adduct formation was observed in the penumbra region during ischemia/reperfusion. This elevation was significantly attenuated by L-NAME during the reperfusion phase. Elevation of free radical adduct formation within the penumbra region during cerebral ischemia/reperfusion may be mediated in part by NOS-dependent mechanisms. PMID- 10799697 TI - Impulsivity and AMPA receptors: aniracetam ameliorates impulsive behavior induced by a blockade of AMPA receptors in rats. AB - The study aimed to ascertain the involvement of central AMPA receptors in impulsive behaviors of aged rats and to examine the effects of aniracetam. Premature response in the two-lever choice reaction task was assessed as an index of impulsivity. Intracerebroventricular injection of 2, 3-dihydroxy-6-nitro-7 sulfamoyl-benzo(F)quinoxaline (NBQX), an AMPA receptor antagonist, dose dependently (10.1-1009 ng/rat) increased only premature response without altering responding speed and choice accuracy 30 min after the injection. Aniracetam (30 mg/kg p.o.), a positive allosteric modulator of AMPA receptors, or AMPA (55.9 ng/rat, co-injected with NBQX) completely restored the NBQX-induced increase in impulsivity. These results indicate that AMPA receptors are tonically involved in the regulation of impulsivity. PMID- 10799698 TI - Excitotoxic profile of LY339434, a GluR5 agonist, in cultured murine cortical neurons. AB - The neurotoxic profile of (2S,4R, 6E)-2-amino-4-carboxy-7-(2-naphthyl)hept-6 enoic acid (LY339434), a low-affinity kainate receptor subtype 5 (GluR5) agonist at recombinant human glutamate receptors, was evaluated to investigate the involvement of GluR5 in excitotoxic neuronal death. Murine cortical neurons were exposed to treatments for 24 h and assessed by a cell viability assay and phase contrast microscopy. LY339434 (1-1000 microM) caused a concentration-dependent decrease in cell viability (EC(50)=11.4+/-1.2 microM) that was only attenuated by (5R, 10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine (MK 801, 10 microM), but not by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 50 microM) or 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine (GYKI 52466, 20 microM). Labeling with nucleic acid binding dyes revealed that LY339434 induced few apoptotic-like characteristics. These findings indicate that in cultured murine cortical neurons, LY339434 acts predominantly through N-methyl D-aspartate (NMDA) receptors rather than GluR5 to effect neuronal death that is rapid and involves predominantly necrosis rather than morphological apoptosis. PMID- 10799699 TI - Effect of chronic intraperitoneal injections of leptin on hypothalamic neurotensin content and food intake. AB - This study was intended for the investigation of the effects of chronic injections of leptin for 7 days on food intake and hypothalamic neurotensin (NT). Leptin treatment significantly reduced food intake [144.3+/-2.5 g (L) vs. 156.7+/ 2.5 g (C); P=0. 002] and body weight gain [23.7 g+/-1.0 g (L) vs. 31.5+/-1.3 g (C); P=0.003]. NT concentration was lower in the lateral hypothalamus (LH) of leptin-treated rats than in the control ad libitum fed rats (-30%; P<0.05). The same diminution was observed in pair-fed rats (-27%; P<0.05). This diminution was therefore related to the decrease in food intake rather than to a direct effect of leptin. As the LH was the only area where NT was modified, it appears that among the hypothalamic nuclei involved in the regulation of feeding behavior it is the most sensitive area to a low energy depletion. Therefore, it might play a specific role in triggering the mechanisms necessary to restore body weight and/or energy balance. PMID- 10799700 TI - Changes in [3H]zolpidem and [3H]Ro 15-1788 binding in rat globus pallidus and substantia nigra pars reticulata following a nigrostriatal tract lesion. AB - Changes in GABA(A) receptor alpha(1) subunit gene expression occur in the globus pallidus and substantia nigra pars reticulata following lesions of the nigrostriatal tract. To determine whether these changes are translated at the protein level, we performed quantitative autoradiography with the alpha(1) selective ligand, [3H]zolpidem, and the non-selective benzodiazepine site ligand, [3H]Ro 15-1788. Binding of both [3H]zolpidem and [3H]Ro 15-1788 was significantly increased in the substantia nigra pars reticulata (13. 5+/-4.1 and 26.3+/-2.9%, respectively) and significantly reduced in the globus pallidus (20.9+/-0.8 and 18.3+/-1.3%, respectively). These changes in alpha(1) subunit protein expression may help to compensate for the pathological changes in GABAergic activity that occur after striatal dopamine depletion. PMID- 10799702 TI - Evidence for the anatomical origins of hypoglossal afferents in the tongue of the leopard frog, Rana pipiens. AB - In this study, the origins of sensory neurons from the tongue that ascend in the hypoglossal nerve were identified and described in the leopard frog, Rana pipiens. Previous studies have shown that these afferents are used to coordinate the timing of jaw and tongue muscles, and are important in the motor control of feeding. These sensory neurons innervate the tongue bilaterally and appear to originate in the dorsal fungiform papillae of the tongue epithelium. PMID- 10799701 TI - Transient depression of excitatory synaptic transmission induced by adenosine uptake inhibition in rat hippocampal slices. AB - The transient property of the dipyridamole-induced depression of excitatory synaptic transmission was analyzed using field EPSPs (fEPSPs) recorded from the CA1 region in rat hippocampal slices. The fEPSPs were depressed by 1 microM dipyridamole and then gradually recovered to the control level. The depression was antagonized by aminophylline or DPCPX, although it was not significantly affected by DMPX. The results suggest that the fEPSP depression is induced by a mechanism through the A(1) receptor. PMID- 10799703 TI - Oxidative stress and HNE conjugation of GLUT3 are increased in the hippocampus of diabetic rats subjected to stress. AB - Recent studies demonstrate that cellular, molecular and morphological changes induced by stress in rats are accelerated when there is a pre-existing strain upon their already compromised adaptive responses to internal or external stimuli, such as may occur with uncontrolled diabetes mellitus. The deleterious actions of diabetes and stress may increase oxidative stress in the brain, leading to increases in neuronal vulnerability. In an attempt to determine if stress, diabetes or stress+diabetes increases oxidative stress in the hippocampus, radioimmunocytochemistry was performed using polyclonal antisera that recognize proteins conjugated by the lipid peroxidation product 4-hydroxy-2 nonenal (HNE). Radioimmunocytochemistry revealed that HNE protein conjugation is increased in all subregions of the hippocampus of streptozotocin (STZ) diabetic rats, rats subjected to restraint stress and STZ diabetic rats subjected to stress. Such increases were not significant in the cortex. Because increases in oxidative stress may contribute to stress- and diabetes-mediated decreases in hippocampal neuronal glucose utilization, we examined the stress/diabetes mediated HNE protein conjugation of the neuron specific glucose transporter, GLUT3. GLUT3 immunoprecipitated from hippocampal membranes of diabetic rats subjected to stress exhibited significant increases in HNE immunolabeling compared to control rats, suggesting that HNE protein conjugation of GLUT3 contributes to decreases in neuronal glucose utilization observed during diabetes and exposure to stress. Collectively, these results demonstrate that the hippocampus is vulnerable to increases in oxidative stress produced by diabetes and stress. In addition, increases in HNE protein conjugation of GLUT3 provide a potential mechanism for stress- and diabetes-mediated decreases in hippocampal neuronal glucose utilization. PMID- 10799704 TI - Recovery of the brain choline level in treated Cushing's patients as monitored by proton magnetic resonance spectroscopy. AB - In a previous study from our group [A. Khiat, C. Bard, A. Lacroix, J. Rousseau, Y. Boulanger, Brain metabolic alterations in Cushing's syndrome as monitored by proton magnetic resonance spectroscopy, NMR Biomed. 12 (1999) 357-363], proton magnetic resonance spectroscopy (1H MRS) was used to evaluate changes in cerebral metabolites in patients with Cushing's syndrome as compared to normal subjects. Data recorded in the frontal, thalamic and temporal areas demonstrated statistically significant decreases of the Cho/Cr ratios in the frontal and thalamic areas but not in the temporal area for Cushing's syndrome patients. No statistically significant changes in the NAA/Cr ratios were measured in any of the areas studied. In this follow-up study, MRS data are reported for ten patients after correction of hypercortisolism which demonstrate a statistically significant recovery of the choline levels in the frontal and thalamic areas. No variation in the NAA, Cr and mI metabolite ratios relative to H(2)O could be measured. Results are interpreted as an inhibition of the phosphatidylcholine degrading phospholipases by glucocorticoids which disappears after correction of hypercortisolism. PMID- 10799705 TI - Telomerase and mammalian ageing: a critical appraisal. AB - The telomeres that occur at the end of chromosomes are maintained by the activity of telomerase and are thought to be important protective factors in maintaining the integrity of chromosomes. It now appears that in vitro replicative senescence, which has been observed in cultured somatic cells, is due to a loss of telomere length in those cells, caused by inactivity of telomerase. This has led to the proposition that telomerase activity is an important determinant in organismal ageing. However, many cells in the body do not proliferate regularly and therefore will not lose telomere length. Cells that do proliferate frequently have now been shown to have active telomerase. Other cells, such as fibroblasts, that do not have telomerase activity but proliferate only occasionally may not reach the Hayflick limit during the lifetime of an animal. There is also no correlation between telomere length and the maximal lifespan exhibited by different species. Studies of telomerase knock-out mice have reported some aspects of accelerated ageing after three generations, but the relevance of these observations to normal ageing remains unconvincing. The role of telomerase in producing immortal tumour cells and the possibility that activation of telomerase is an important event in malignant transformation is similarly controversial and open to alternative interpretations. The significance of these and other observations, and how they define the role of telomerase in ageing, is discussed. PMID- 10799706 TI - Increased TNF-alpha and PGI(2), but not NO release from macrophages in 18-month old rats. AB - TNF-alpha is an important pro-inflammatory mediator that influences host defense against infection and cancer. Previous examinations of TNF-alpha release and synthesis within the context of age have provided conflicting data, as both increased and decreased TNF-alpha synthesis have been described in aged populations. The present study was designed to reevaluate TNF-alpha production and synthesis in primary cultured peritoneal macrophages of young and 18-month old rats. We were also interested in the link between the production of this cytokine and other important mediators, such as prostaglandin I(2) (PGI(2)) and nitric oxide (NO) in these rats. Primary cultured peritoneal macrophages of rat were stimulated with 1.0 microg/ml of lipopolysaccharide (LPS) for 12 h. The level of TNF-alpha protein in culture supernatant was measured by enzyme-linked immunosorbent-assay (ELISA), and TNF-alpha mRNA production was assessed by semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). At the same time, the levels of NO and PGI(2) were measured. Macrophages from 18-month old rats produced more TNF-alpha protein, PGI(2) and TNF-alpha mRNA than those from the young rats (2 month). There was no difference in NO production of macrophages between 18-month-old and young rats. The results demonstrate that TNF alpha and PGI(2) production by rat macrophages increase with age. The results also suggest that NO might not contribute to the increased TNF-alpha production in 18-month-old rat macrophages. PMID- 10799707 TI - Tubular aggregates in the skeletal muscle of the senescence-accelerated mouse; SAM. AB - We investigated the skeletal muscles of nine strains of senescence accelerated mouse (SAM), DDD, AKR/J, C57BL/6J, A/J and BALB/c mice. We found that male SAMP8, SAMP7, C57BL/6J, A/J and BALB/c mice expressed tubular aggregates (TAs) in their skeletal muscle. Among these strains, the SAMP8 strain, which exhibits a short life span and various age-associated neurodegenerative disorders plus mitochondrial dysfunction, showed TAs more markedly than the others. Thus, we compared SAMP8 mice against SAMR1 mice, an accelerated senescence-resistant strain. Light- and electron micrographs showed that male SAMP8 mice exhibited an age-dependent aggravation of TA accumulation. There were no significant differences in the serum lactate/pyruvate levels between the SAMP8 and SAMR1 mice. However, the serum creatine kinase (CK) levels of the 3 and 6-month-old SAMP8 mice were higher than that of the corresponding SAMR1 mice. Considering the serum CK levels and the mitochondrial dysfunction of SAMP8 mice, we conclude that the TAs may be involved in the homeostasis of energy metabolism that is not appropriately regulated in the SAMP8 mouse mitochondrion. PMID- 10799708 TI - Altered aging-related thymic involution in T cell receptor transgenic, MHC deficient, and CD4-deficient mice. AB - During aging in mice and humans, a gradual decline in thymus integrity and function occurs (thymic involution). To determine whether T cell reactivity or development affects thymic involution, we compared the thymic phenotype in old (12 months) and young (2 months) mice transgenic for rearranged alphabeta or beta 2B4 T cell receptor (TCR) genes, mice made deficient for CD4 by gene targetting (CD4(-/-)), mice made deficient for major histocompatibility complex (MHC) class I (beta2M-/-) or class II genes (A(beta)(b-/-) on C57Bl/6 background) or both. The expected aging-related reductions in thymic weights were observed for all strains except those bearing disruption of both class I and class II MHC genes. Therefore, disruption of MHC class I and class II appeared to reverse or delay aging-related thymic atrophy at 12 months. Immunohistochemical analysis of aging associated alterations in thymic morphology revealed that TCR alphabeta transgenes, CD4 disruption, and MHC class II disruption all reduced or eliminated these changes. All strains examined at 12 months showed alterations in the distribution of immature thymocyte populations relative to young controls. These results show that aging-associated thymic structural alterations, size reductions, and thymocyte developmental delays can be separated and are therefore causally unrelated. Furthermore, these results suggest that the T cell repertoire and/or its development play a role in aging-related thymic involution. PMID- 10799709 TI - Tissue specific expression of PKR protein kinase in aging B6D2F1 mice. AB - A decline in the rate of protein synthesis is a common biochemical change observed with aging in a wide variety of cells and organisms. The double stranded RNA-dependent protein kinase PKR has been shown to phosphorylate eukaryotic initiation factor 2 alpha (eIF-2alpha), a well-characterized factor for down regulating protein synthesis, in response to environmental stress conditions. Therefore, we were interested in evaluating the role of PKR in the aging process. Tissues from 2- and 20-month-old B6D2F1 male mice were evaluated by Western blot analysis. PKR was detected in all tissues of aging mice confirming its ubiquitous nature. Tissues examined from young mice showed little evidence of PKR expression, suggesting an age-associated up-regulation. P58(IPK), a cellular inhibitor of PKR, was expressed in tissues from both age groups but to a greater extent in tissues of aging mice suggesting an up-regulation to control PKR activity. Hyperphosphorylated eIF-2alpha was increased in selected tissues from older mice compared with tissues from younger mice indicating a possible correlation between PKR expression and kinase function. The data suggest that translational activity is slowing down in a tissue specific manner during the aging process in mice, possibly as the result of increased levels of PKR, and could be a factor in the reduction of the rate of protein synthesis during senescence seen in specific tissues of many organisms. PMID- 10799710 TI - Calorie restriction modulates age-dependent changes in the retinas of Brown Norway rats. AB - The present study examined the effect of a 40% reduction in caloric intake (CR) versus ad libitum (AL) feeding on retinal aging. CR- and AL-fed Brown Norway (BN) rats were obtained at 12, 24 and 30 months of age from the National Center for Toxicological Research (NCTR). Age-dependent declines in outer nuclear layer (ONL=photoreceptor) cell densities, ONL height, inner nuclear layer (INL) cell densities, and thicknesses of the inner retina and whole retina were quantified in thick sections at six loci across the circumference of the sensory retina (four peripheral, two central). Data were analyzed by repeated measures, general linear models. Aging in both diet groups was associated with declines in ONL cell density, ONL height, peripheral INL cell density and total retinal thickness (P< or =0.05). However, ONL cell densities, ONL height and retinal thickness were significantly greater in the CR versus AL diet group at all three ages (P< or =0.005). CR was also associated with a trend for greater peripheral INL cell density (P=0.06) and with greater INL thickness at 30 months (Bonferroni P=0.03). Elevated ONL cell densities in the CR-12 cohort relative to the AL-12 cohort could be explained by diet-associated differences in retinal length, i.e. delayed retinal growth in response to CR. Enhanced ONL cell density, ONL height, INL cell density, INL thickness and total retinal thickness in the CR-30 cohort appear to be as a result of reduced rates of retinal cell loss between 24 and 30 months. However, the protective effect of CR in retinas of older animals may also reflect the initial growth-associated enhancements which were observed in 12 month-old animals. The rat retina may provide a useful model for elucidating the neuroprotective mechanism(s) of CR. PMID- 10799711 TI - Factors influencing the processing and function of the amyloid beta precursor protein--a potential therapeutic target in Alzheimer's disease? AB - The amyloid beta precursor protein (AbetaPP), which plays a pivotal role in Alzheimer's disease (AD), can exist as either a membrane-bound or soluble protein. The former is cleaved at the level of the plasma membrane to generate the soluble form of the protein (AbetaPP(s)). An alternative pathway exists, however, for the cleavage of AbetaPP to generate a 40-42 amino acid peptide termed amyloid beta (Abeta), either within the lysosomal or the endoplasmic reticulum/Golgi compartments of the cell. In AD, there is an increase in the ratio of the 42 amino acid form of the Abeta peptide (Abeta(42)) to Abeta(40). The Abeta(42) form is the more amyloidogenic form and has an increased potential to form the insoluble amyloid deposits characteristic of AD pathology. Studies on the familial form of the disease, with mutations in AbetaPP or in the presenilin proteins, have confirmed an increase in Abeta(42) generation associated with the early stages of the disease. This review will examine the factors that influence AbetaPP processing, how they may act to modulate the biological effects of AbetaPP(s) and Abeta, and if they provide a viable target for therapeutic intervention to modify the rate of progression of the disease. PMID- 10799712 TI - Human immunodeficiency virus type-1 protease inhibitors: therapeutic successes and failures, suppression and resistance. AB - The retroviral protease (PR) is responsible for cleaving precursor proteins that contain the virion structural proteins and enzymes. Highly potent inhibitors of the human immunodeficiency virus type-1 PR have been developed, and to date, five of these inhibitors have been approved for clinical use. These inhibitors bind to the active site of the dimeric PR and represent transition state analogs. Combination therapy in which a potent protease inhibitor is combined with inhibitors of the viral DNA polymerase reverse transcriptase can result in the apparent complete suppression of virus replication. Low virus loads associated with suppressed replication are resulting in dramatic reductions in the rate of disease progression. However, incomplete suppression of virus replication results in the selection of resistant variants. Resistance to protease inhibitors is the result of mutations within the PR coding domain, and most of these mutations are able to contribute to cross-resistance among this class of inhibitors. PMID- 10799713 TI - Neuronal fast oscillations as a target site for psychoactive drugs. AB - Neuronal oscillations within the electroencephalogram beta and gamma bands (15-80 Hz) are associated with intense mental activity and cognitive function in general. Specifically, recent advances have implicated gamma oscillations in the processing of sensory stimuli and demonstrated that synchronous gamma oscillations, appearing concurrently in spatially separate brain regions, can induce beta activity. beta activity generated in this manner represents established synchronous communication between brain regions and is thought to represent a neuronal network correlate of the "binding phenomenon" in cognitive theory. This review will outline the mechanisms of generation of these oscillations at the cellular and network level, and will highlight the effects of drugs that may modify these mechanisms. Possible modification of fast oscillations by disease processes and clinical intervention are discussed. PMID- 10799714 TI - Traditional Chinese medicine: an approach to scientific proof and clinical validation. AB - The classical Chinese pharmacopoeia describes a large number of herbal formulations that are used for the treatment of a wide variety of diseases. This therapeutic approach is ignored by many and considered to be an alternative to conventional medicine by others. The scientific proof and clinical validation of these herbal formulations require a rigorous approach that includes chemical standardization, biological assays, animal models, and clinical trials. Such Western methodologies need to take into consideration the complex mixture of chemicals and how they are to be used in humans. This review examines relevant studies on the use of traditional Chinese medicines for the treatment of such diseases as bronchial asthma, atopic dermatitis, and irritable bowel syndrome. An interdisciplinary approach to traditional Chinese medicine may provide a platform for the discovery of novel therapeutics composed of multiple chemical compounds. PMID- 10799716 TI - Sphingomyelin metabolites in vascular cell signaling and atherogenesis. AB - The atherosclerotic lesion most probably develops through a number of cellular events which implicate all vascular cell types and include synthesis of extracellular proteins, cell proliferation, differentiation and death. Sphingolipids and sphingolipid metabolizing enzymes may play important roles in atherogenesis, not only because of lipoprotein alterations but also by mediating a number of cellular events which are believed to be crucial in the development of the vascular lesions such as proliferation or cell death. Exogenous sphingolipids may mediate various biological effects such as apoptosis, mitogenesis or differentiation depending on the cell type. Moreover, several molecules present in the atherogenic lesion, such as oxidized LDL, growth factors or cytokines, which activate intracellular signaling pathways leading to vascular cell modifications, can stimulate sphingomyelin hydrolysis and generation of ceramide (and other metabolites as sphingosine-1-phosphate). Here we review the potential implication of the sphingomyelin/ceramide cycle in vascular cell signaling related to atherosclerosis, and more generally the role of sphingolipids in the events observed during the atherosclerotic process as cell differentiation, migration, adhesion, retraction, proliferation and death. PMID- 10799717 TI - Vitamin E: non-antioxidant roles. AB - Vitamin E was originally considered a dietary factor of animal nutrition especially important for normal reproduction. The significance of vitamin E has been subsequently proven as a radical chain breaking antioxidant that can protect the integrity of tissues and play an important role in life processes. More recently alpha-tocopherol has been found to possess functions that are independent of its antioxidant/radical scavenging ability. Absorption in the body is alpha-tocopherol selective and other tocopherols are not absorbed or are absorbed to a lesser extent. Furthermore, pro-oxidant effects have been attributed to tocopherols as well as an anti-nitrating action. Non-antioxidant and non-pro-oxidant molecular mechanisms of tocopherols have been also described that are produced by alpha-tocopherol and not by beta-tocopherol. alpha Tocopherol specific inhibitory effects have been seen on protein kinase C, on the growth of certain cells and on the transcription of some genes (CD36, and collagenase). Activation events have been seen on the protein phosphatase PP2A and on the expression of other genes (alpha-tropomyosin and Connective Tissue Growth Factor). Non-antioxidant molecular mechanisms have been also described for gamma-tocopherol, delta-tocopherol and tocotrienols. PMID- 10799718 TI - The biosynthesis and functional role of cardiolipin. PMID- 10799719 TI - In vitro radiosensitivity and normal tissue damage. PMID- 10799720 TI - Is there a role for molecular prognostic factors in the clinical management of ductal carcinoma in situ (DCIS) of the breast? AB - INTRODUCTION: The incidence of ductal carcinoma of the breast (DCIS) increased in Europe and the US up to 10-fold over the last 20 years ?8. This could be linked to more vigorous screening mammography, as well as changes of histopathologic and diagnostic criteria for breast lesions during the last decades ?31,26. Optimal screening for DCIS, the diagnostic procedures and best treatment is still controversial. For many DCIS patients lumpectomy and adjuvant radiotherapy are a valid treatment option. There is need for better prognostic factors in DCIS, which indicate the need for therapy and tailor the intensity of treatment. Recently prognostic factors based on clinical and pathological findings for DCIS were established and are currently validated. Molecular mechanisms involved in DCIS formation, DCIS progression to invasive breast cancer, and predicting DCIS treatment response are rapidly emerging ?46. RESULTS AND CONCLUSION: Here we discuss some of the known molecular mechanisms of DCIS and how they could be further exploited as prognostic factors for screening and tailoring DCIS therapy. This review will summarize relevant molecular mechanism of DCIS carcinogenesis including dysregulation of the cell cycle clock and changes of the apoptotic threshold. In particular, recently published molecular and cellular abnormalities in DCIS, potentially relevant for treatment decision, will be discussed. PMID- 10799721 TI - Randomized clinical trial on 7-day-continuous accelerated irradiation (CAIR) of head and neck cancer - report on 3-year tumour control and normal tissue toxicity. AB - PURPOSE: To evaluate tumour and normal tissues 3-year response to 7-day-a-week continuous accelerated irradiation (CAIR) compared to a conventional treatment (5 days per week) in a randomized trial. MATERIALS AND METHODS: One hundred patients with squamous cell carcinoma of the head and neck in stage T(2-4)N(0-1)M(0) were entered into the trial between December 1, 1993 and June 30, 1996. Dose per fraction of 2.0 Gy (to the end of 1994), and 1.8 Gy (since January 1, 1995) was the same in both arms and delivered once a day at regular 24-h intervals to total dose in the range of 66-72 Gy (depending on tumour stage). The only difference was overall treatment time being 5 weeks in the CAIR and 7 weeks in control arm. RESULTS: Actuarial 3-year local tumour control was 82% in the CAIR and 37% in the control group (P<0.0001) with reduction in local recurrence rate of 83%. Actuarial 3-year overall survival was 78 and 32% (P<0.0001), respectively. Confluent mucositis was significantly more severe and lasted longer in the CAIR than in control arm. After 2.0 Gy fractions five of 23 patients (22%) in the CAIR developed early necroses over a period of 2-4 months of follow-up which can be considered as a consequential to severe protracted acute mucosal reactions (CLE). For this reason dose per fraction was lowered to 1. 8 Gy and the CLE was not observed again until now. Thus the overall rate of CLE decreased to 10%. CONCLUSIONS: The gain in tumour control is likely the effect of shortening of overall treatment time by 14 days and regular continuous dose delivery during the whole course of radiation therapy including weekends. A 7-day schedule produces more severe acute mucosal reactions lasting longer than in conventional fractionation, however tolerable by patients. Relatively high rate (22%) of CLE in the 7-day arm observed during the first year of the study was eliminated by decreasing dose per fraction from 2.0 Gy to 1.8 Gy. PMID- 10799722 TI - ARCON: accelerated radiotherapy with carbogen and nicotinamide in head and neck squamous cell carcinomas. The experience of the Co-operative group of radiotherapy of the european organization for research and treatment of cancer (EORTC). AB - BACKGROUND: Since there is increasing evidence that both acute (perfusion limited) and chronic (diffusion-limited) hypoxia, and tumor repopulation may prejudice the outcome of radiotherapy, the combination of carbogen (95% oxygen-5% carbon dioxide) and nicotinamide with accelerated radiotherapy (ARCON) should reduce the impact of these factors of radioresistance. AIM: This clinical study was aimed at determining the feasibility, as well as the qualitative and quantitative toxic effects of a therapeutic approach based on ARCON, and assessing the tumor response rates that can be achieved with this regime in patients with locally advanced tumors of the head and neck. METHODS: A phase I/II study conducted between 1993 and 1996 by the Co-operative Group of Radiotherapy of the EORTC included three consecutive steps: accelerated fractionation (AF) combined with carbogen (11 analyzable patients), AF combined with the daily administration of nicotinamide (n=10), and AF with both carbogen and nicotinamide (n=17). Radiotherapy was based on an accelerated regime (72 Gy in 5.5 weeks). Nicotinamide was delivered 90 min before the first irradiation session, at a daily dose of 6 g. Carbogen breathing started 5 min before irradiation and lasted throughout the entire radiotherapy sessions. RESULTS: No significant difference in loco-regional toxicity was found among the three study steps, when carbogen and nicotinamide, either alone or in combination, were combined with AF. The feasibility of the ARCON protocol, as proposed in the present EORTC study, appears to be significantly impaired when nicotinamide is added, at a daily dose of 6 g, to AF and carbogen, in an unselected group of patients. More than 20% of patients experienced grade 2 or 3 emesis. It also demonstrates, in unselected groups of patients, no significant difference in tumor response and local control when carbogen and nicotinamide, either alone or in combination, are added to accelerated radiotherapy. The percentages of objective response at 2 months were 81, 70 and 87%, respectively. CONCLUSION: Future ARCON trials should target selected head and neck tumor localizations and stages, and a lower nicotinamide dose is needed to reduce severe upper gastro-intestinal toxicity. PMID- 10799723 TI - Cervical lymph node metastases from unknown primary tumours. Results from a national survey by the Danish Society for Head and Neck Oncology. AB - BACKGROUND AND PURPOSE: The management of patients with cervical lymph node metastases from unknown primary tumours is a major challenge in oncology. This study presents data collected from all five oncology centres in Denmark. MATERIAL AND METHODS: Of the 352 consecutive patients with squamous cell or undifferentiated tumours seen from 1975 to 1995, a total of 277 (79%) were treated with radical intent. The general treatment policy at all centres during the entire study period has been to treat all suitable candidates with radiotherapy to both sides of the neck and include elective irradiation of the mucosal sites in nasopharynx, and larynx, hypopharynx and larynx (81%). Irradiation of the ipsilateral neck only was done in 26 patients (10%). Radical surgery was the only treatment in 23 N1-N2 patients (9%). RESULTS: The 5-year estimates of neck control, disease-specific survival and overall survival for radically treated patients were 51, 48 and 36%, respectively. The emergence of the occult primary was observed in 66 patients (19%). About half of the emerging primaries were within the head and neck region with oropharynx, hypopharynx and oral cavity being the most common sites. Emerging primaries outside the head and neck region were primarily located in the lung (19 patients) and oesophagus (five patients). The frequency of emerging primary in the head and neck was significantly higher in patients treated with surgery alone, the actuarial risks at 5-year being 54+/-1% (no RT) vs. 15+/-3% (with RT), P<0.0001. The most important factor for neck control was nodal stage (5-year estimates 69% (N1), 58% (N2) and 30% (N3)). Other important parameters for neck control and disease specific survival included haemoglobin, gender and overall treatment time. Patients treated with ipsilateral radiotherapy had a relative risk of recurrence in the head and neck region of 1.9 compared with patients treated to both neck and mucosa. At 5 years, the estimated control rates were 27% (ipsilateral) and 51% (bilateral; P=0.05). The 5-year disease-specific survival estimates were 28 and 45%, respectively (P=0.10). CONCLUSIONS: This study has confirmed that patients with neck node metastases from occult head and neck cancer have clinical features and prognosis similar to other head and neck malignancies. Extensive irradiation to both sides of the neck and the mucosa in the entire pharyngeal axis and larynx resulted in significantly less loco-regional failures compared with patients treated with ipsilateral techniques, but only a trend towards better survival. A prospective randomized trial is required to determine the optimal strategy in terms of locoregional control, survival and morbidity. PMID- 10799724 TI - CT-based parotid gland location: implications for preservation of parotid function. AB - PURPOSE: The position of the parotid gland in relation to surrounding structures was investigated. MATERIALS AND METHODS: Sixty-five patients with head and neck tumours were prospectively evaluated. Parotid position was determined using beam's eye views of CT images projected on simulator films. Distances between the different borders of the parotid gland and surrounding bony marks were quantitatively assessed. RESULTS: The parotid gland volume ranged from 12.9 to 46.4 cm(3). The distance between the cranial border of the parotid gland and the tuberculum anterior of the atlas ranged between 0.7 and 4.8 cm. The position of the parotid gland was unaffected by the angle of the mandible. CONCLUSIONS: The size and position of the parotid gland varies largely among patients. As the extent of radiation-induced salivary dysfunction depends on the volume of the gland tissue exposed, CT-based simulation of radiation fields is necessary. PMID- 10799725 TI - Local tumor control and morbidity after one to three fractions of stereotactic external beam irradiation for uveal melanoma. AB - BACKGROUND AND PURPOSE: To evaluate prospectively local tumor control and morbidity after 1-3 fractions of stereotactic external beam irradiation (SEBI) in patients with uveal melanoma, unsuitable for ruthenium-106 brachytherapy or local resection. MATERIAL AND METHODS: This phase I/II study includes 62 selected patients with uveal melanoma. The mean initial tumor height was 7.8+/-2.8 mm. With the Leskell gamma knife SEBI, 41 patients (66%) were irradiated with two equal fractions of 35, 30 or 25 Gy/fraction, 14 patients (22%) were treated with three fractions of 15 Gy each, and seven patients (11%) with small tumor volumes below 400 mm(3) were treated with one fraction of 45 Gy. The mean total dose was 54+/-8 Gy. The minimal follow-up period was 12 months, and the median follow-up was 28.3 months. Data on radiation-induced side-effects were analyzed with the Cox proportional hazards model for possible risk factors. RESULTS: Local tumor control was achieved in 98% and tumor height reduction in 97%. The mean relative tumor volume reductions were 44, 60 and 72% after 12, 24 and 36 months, respectively. Seven patients developed metastases (11%). Secondary enucleation was performed in eight eyes (13%). Morbidity was significant in tumors exceeding 8 mm in initial height; it was comparable and acceptable in those smaller. In the stepwise multiple Cox model, tumor localization, height and volume, planning target volume (PTV), total dose and patient age were identified as the strongest risk factors for radiation-induced lens opacities, secondary glaucoma, uveitis, eyelash loss and exudative retinal detachment. In this model, the high-dose volume irradiated with more than 10 Gy/fraction was the strongest risk factor for radiation-induced uveitis. CONCLUSIONS: Stereotactic external photon beam irradiation and a total dose of 45-70 Gy delivered in one to three fractions are highly effective at achieving local tumor control in uveal melanoma. Further clinical studies using smaller fraction doses, and consequent smaller high-dose volumes, are justified to optimize dose and fractionation. Fractionated stereotactic irradiation has a challenging potential as an eye-preserving treatment in uveal melanoma. PMID- 10799726 TI - Cardiac and lung complication probabilities after breast cancer irradiation. AB - PURPOSE: To assess for locoregional irradiation of breast cancer patients, the dependence of cardiac (cardiac mortality) and lung (radiation pneumonitis) complications on treatment technique and individual patient anatomy. MATERIALS AND METHODS: Three-dimensional treatment planning was performed for 30 patients with left-sided breast cancer and various breast sizes. Two locoregional techniques (Techniques A and B) and a tangential field technique, including only the breast in the target volume, were planned and evaluated for each patient. In both locoregional techniques tangential photon fields were used to irradiate the breast. The internal mammary (IM)-medial supraclavicular (MS) lymph nodes were treated with an anterior mixed electron/photon field (Technique A) or with an obliquely incident mixed electron/photon IM field and an anterior electron/photon MS field (Technique B). The optimal IM and MS electron field dimensions and energies were chosen on the basis of the IM-MS lymph node target volume as delineated on CT-slices. The position of the tangential fields was adapted to match the IM-MS fields. Dose-volume histograms (DVHs) and normal tissue complication probabilities (NTCPs) for the heart and lung were compared for the three techniques. In the beam's eye view of the medial tangential fields the maximum distance of the heart contour to the posterior field border was measured; this value was scored as the Maximum Heart Distance. RESULTS: The lymph node target volume receiving more than 85% of the prescribed dose was on average 99% for both locoregional irradiation techniques. The breast PTV receiving more than 95% of the prescribed dose was generally smaller using Technique A (mean: 90%, range: 69-99%) than using Technique B (mean: 98%, range: 82-100%) or for the tangential field technique (mean: 98%, range: 91-100%). NTCP values for excess cardiac mortality due to acute myocardial ischemia varied considerably between patients, with minimum and maximum values of 0.1 and 7.5% (Technique A), 0.1 and 5.8% (Technique B) and 0.0 and 6.1% (tangential tech.). The NTCP values were on average significantly higher (P<0.001) by 1.7% (Technique A) and 1.0% (Technique B) when locoregional breast irradiation was given, compared with irradiation of the left breast only. The NTCP values for the tangential field technique could be estimated using the Maximum Heart Distance. NTCP values for radiation pneumonitis were very low for all techniques; between 0.0 and 1.0%. CONCLUSIONS: Technique B results in a good coverage of the breast and locoregional lymph nodes, while Technique A sometimes results in an underdosage of part of the target volume. Both techniques result in a higher probability of heart complications compared with tangential irradiation of the breast only. Irradiation toxicity for the lung is low in all techniques. The Maximum Heart Distance is a simple and useful parameter to estimate the NTCP values for cardiac mortality for tangential breast irradiation. PMID- 10799727 TI - Long-term cardiac mortality following radiation therapy for Hodgkin's disease: analysis with the relative seriality model. AB - PURPOSE: (a) To assess the increased risk of death due to ischemic heart disease (IHD) in a group of patients treated for Hodgkin's disease (HD) with radiation therapy (RT) as the primary treatment. (b) To quantify the dose response of IHD using a biophysical model. MATERIALS AND METHODS: Patient material consisted of 157 patients diagnosed for HD between 1972 and 1985 who received RT as the primary treatment at Radiumhemmet, Karolinska Hospital. The general population formed the control group. The RT treatments were reconstructed based on the individual treatment data and simulator films. Individual clinical and dosimetrical data were analyzed with the relative seriality model. The material was also analyzed grouping the material according to dose-volume constraints. RESULTS: Of the 157 patients, 13 (8.3%) died due to IHD. The standardized mortality ratio (SMR) was 5.0 (95% CI, 2.7-8.6). Analysis of dose-volume histograms (DVH) showed an increasing risk with increasing dose to a larger volume fraction. The observed individual clinical complication data could not be modeled unambiguously. The group analysis resulted in the dose-response parameters: D(50)=71 Gy, gamma=0.96 and s=1.0. CONCLUSIONS: A significantly increased risk of death due to IHD following RT for HD was found. The risk was found to increase with higher dose and larger volume fraction irradiated. PMID- 10799728 TI - Myocardial perfusion imaging in breast cancer patients treated with or without post-mastectomy radiotherapy. AB - PURPOSE: To assess the occurrence and location of myocardial perfusion defects in left-sided mastectomized breast cancer patients, treated with or without postoperative radiotherapy according to the guidelines from the Danish Breast Cancer Cooperative Group (DBCG). PATIENTS AND METHODS: Seventeen left-sided breast cancer patients, with a median age of 59 years (range, 47-75 years), randomized to post-mastectomy irradiation plus systemic treatment, or systemic treatment alone, were examined after a median follow-up of 7.9 years (range, 6.0 12.2 years). The chest wall and the ipsilateral internal mammary nodes had been treated through two anterior-shaped electron fields, and the electron energy was chosen according to chest wall thickness, measured individually by ultrasound. The median absorbed dose was 50 Gy in 25 fractions, with 5 fractions/week. Information on clinical history was obtained and symptoms of ischemic heart disease (IHD), as well as major risk factors, were recorded. All patients had a physical examination, blood chemistry, electrocardiogram (ECG), chest X-ray and myocardial perfusion imaging by sestamibi-single photon emission computerized tomography (SPECT). SPECT-scanning was performed as a rest/dipyridamole 2-day protocol. The evaluation of regional myocardial perfusion was based on scintigrams using a 20-segment model. RESULTS: There was no significant difference between the scintigraphic findings in the two groups. Four of ten irradiated patients and four of seven non-irradiated patients showed scintigraphic defects. An anterior defect was found in one non-irradiated patient. CONCLUSIONS: This study does not indicate that the described radiotherapy technique induces detectable coronary artery disease. However, the small number of patients does not allow strong conclusions to be drawn. PMID- 10799729 TI - Cellular radiosensitivity and complication risk after curative radiotherapy. AB - PURPOSE: To test for an association between in vitro fibroblast radiosensitivity and complication risk in a case-control study of breast cancer patients treated under standard conditions in a clinical trial of radiotherapy dose fractionation. PATIENTS AND METHODS: A cohort of patients participating in a randomised clinical trial of radiotherapy dose fractionation was selected on the basis of treatment induced changes in the breast several years later. Thirty-nine cases with marked normal tissue changes were matched on several variables with 65 controls with no changes attributable to radiotherapy. Dermal fibroblast strains were established from duplicate skin biopsies, and clonogenic cell survival assays performed in triplicate after both high ( approximately 1.6 Gy/min) and low ( approximately 1 cGy/min) dose-rate irradiation. Laboratory studies were blind to patient identity, treatment outcome and radiotherapy schedule. RESULTS: Analysis of 1128 clonogenic survival curves confirmed significant inter-patient variation in fibroblast radiosensitivity as measured by clonogenic survival. However, no association between fibroblast radiosensitivity and the development of late radiotherapy normal tissue effects was detected. CONCLUSIONS: Inter-individual variation in cellular radiosensitivity may not be the main determinant of complication risk in patients undergoing radiotherapy for breast cancer. Other biological and technical factors may be more important in explaining the marked inter-patient differences in normal tissue damage evident several years after curative radiotherapy. PMID- 10799730 TI - Relationship between in vitro chromosomal radiosensitivity of peripheral blood lymphocytes and the expression of normal tissue damage following radiotherapy for breast cancer. AB - BACKGROUND AND PURPOSE: There is a need for rapid and reliable tests for the prediction of normal tissue responses to radiotherapy, as this could lead to individualization of patient radiotherapy schedules and thus improvements in the therapeutic ratio. Because the use of cultured fibroblasts is too slow to be practicable in a clinical setting, we evaluated the predictive role of assays of lymphocyte chromosomal radiosensitivity in patients having radiotherapy for breast cancer. MATERIALS AND METHODS: Radiosensitivity was assessed using a micronucleus (MN) assay at high dose rate (HDR) and low dose rate (LDR) on lymphocytes irradiated in the G(0) phase of the cell cycle (Scott D, Barber JB, Levine EL, Burril W, Roberts SA. Radiation-induced micronucleus induction in lymphocytes identifies a frequency of radiosensitive cases among breast cancer patients: a test for predispostion? Br. J. Cancer 1998;77;614-620) and an assay of G(2) phase chromatid radiosensitivity ('G(2) assay') (Scott D, Spreadborough A, Levine E, Roberts SA. Genetic predisposition in breast cancer. Lancet 1994; 344: 1444). In a study of acute reactions, blood samples were taken from breast cancer patients before the start of radiotherapy, and the skin reaction documented. 116 patients were tested with the HDR MN assay, 73 with the LDR MN assay and 123 with the G(2) assay. In a study of late reactions, samples were taken from a series of breast cancer patients 8-14 years after radiotherapy and the patients assessed for the severity of late effects according to the'LENT SOMA' scales. 47 were tested with the HDR assay, 26 with the LDR assay and 19 with the G(2) assay. For each clinical endpoint, patients were classified as being normal reactors or 'highly radiosensitive patients' (HR patients (Burnet NG. Johansen J, Turesson I, Nyman J. Describing patients' normal tissue reactions: Concerning the possiblity of individualising radiotherapy dose presciptions based on potential predictive assays of normal tissue radiosensitivity. Int. J. Cancer 1998;79:606-613)). RESULTS: The HR patients could be identified in some of the assays. For example, for acute skin reactions, 9/123 patients were judged as HR; they had significantly higher G(2) scores than normal reactors (P=0.004). For the late reactions, the mean HDR MN scores were higher for the 4/47 patients who had severe telangiectasia (P=0.042) and the 8/47 patients had severe fibrosis (P=0.055). However, there were no trends towards increased chromosomal radiosensitivity with the micronucleus scores at HDR or LDR, or with G(2) chromosomal radiosensitivity. CONCLUSIONS: While these results support the concept of using lymphocytes to detect elevated sensitivity to radiotherapy (as an alternative to fibroblasts), these assays are unlikely to be of assistance for the prediction of normal tissue effects in the clinic in their present form. PMID- 10799731 TI - Radiobiological depth of subcutaneous induration. AB - The incidence of subcutaneous induration in 23 breast cancer patients with postmastectomy radiotherapy was investigated. The patients were treated with three different radiotherapy schedules 7. 9-9.8 years ago. The incidence was correlated with the biological effective dose (BED) at different depths from the skin surface and a best fit was obtained using an alpha/beta ratio of 2 Gy. The most representative depth at which to assess dose for subcutaneous induration was 2 mm which is near the subcutaneous fat/dermis interface. PMID- 10799732 TI - Skin damage probabilities using fixation materials in high-energy photon beams. AB - INTRODUCTION: Patient fixation, such as thermoplastic masks, carbon-fibre support plates and polystyrene bead vacuum cradles, is used to reproduce patient positioning in radiotherapy. Consequently low-density materials may be introduced in high-energy photon beams. The aim of the this study was to measure the increase in skin dose when low-density materials are present and calculate the radiobiological consequences in terms of probabilities of early and late skin damage. METHOD: An experimental thin-windowed plane-parallel ion chamber was used. Skin doses were measured using various overlaying low-density fixation materials. A fixed geometry of a 10x10 cm field, a SSD=100 cm and photon energies of 4, 6 and 10 MV on Varian Clinac 2100C accelerators were used for all measurements. Radiobiological consequences of introducing these materials into the high-energy photon beams were evaluated in terms of early and late damage of the skin based on the measured surface doses and the LQ-model. RESULTS: The experimental ion chamber gave results consistent with other studies. A relationship between skin dose and material thickness in mg/cm(2) was established and used to calculate skin doses in scenarios assuming radiotherapy treatment with opposed fields. CONCLUSION: Conventional radiotherapy may apply mid-point doses up to 60-66 Gy in daily 2-Gy fractions opposed fields. Using thermoplastic fixation and high-energy photons as low as 4 MV do increase the dose to the skin considerably. However, using thermoplastic materials with thickness less than 100 mg/cm(2) skin doses are comparable with those produced by variation in source to skin distance, field size or blocking trays within clinical treatment set-ups. The use of polystyrene cradles and carbon-fibre materials with thickness less than 100 mg/cm(2) should be avoided at 4 MV at doses above 54-60 Gy. PMID- 10799733 TI - Pregnancy induced mammary tumor specific effector cells are present long after parturition in a breast cancer model in rats. AB - Pregnancy is known to provide protection against 7,12 dimethylbenz[a]anthracene (DMBA) induced mammary carcinogenesis in rats. We observed in earlier studies that splenocytes of parous rats have significant cytotoxicity against mammary tumor cells both in vitro and in vivo. However, it remains to be established how long these cytolytic cells persist after parturition in parous host. The present study was designed using parous rats, 36 or more days after parturition. We observed that fresh splenocytes from these rats had low cytolytic activity against mammary tumor cells. However, when these cells were re-stimulated with irradiated mammary tumor cells in vitro, they had significantly higher cytotoxicity against mammary tumor cells. These studies show for the first time that pregnancy induced cytotoxic splenocytes are present long after parturition and they can be restimulated in vitro to enhance the cytotoxic effect. PMID- 10799734 TI - Evidence for microsatellite instability in bilateral breast carcinomas. AB - The molecular pathogenesis of various categories of breast cancer (BC) has been well described, but surprisingly few reports have appeared on analysis of somatic mutations in bilateral BC. We have performed a polymerase chain reaction (PCR) driven investigation of chromosomal regions showing common loss of heterozygosity (LOH) in 23 cases (46 tumors) from patients diagnosed with bilateral BC. LOH was observed in 15/46 (33%) informative tumors for chromosome 1p, 5/32 (16%) for 5q, 12/44 (27%) for 11q, 15/40 (38%) for 13q and 4/24 (17%) for 17p. These values are within the range of interlaboratory variations reported for unilateral BC. There was no strong evidence for concordance of LOH within the same patient for any of the chromosomal loci tested. Atypical for breast carcinomas, 7/46 (15%) tumors accumulated a high frequency (ranging from 11 to 29%) of shortened dinucleotide CA repeats, implying microsatellite instability (MI). Further analysis with the highly informative BAT-26 marker allowed for the classification of two of these tumors as having a replication error positive (RER(+)/MSI-H) phenotype, whereas the remaining five carcinomas harbored so-called borderline MI. Thus an involvement of both RER(+) and borderline MI appears to be a distinct feature of bilateral breast carcinomas compared to unilateral lesions. PMID- 10799735 TI - Fatty acyl-CoAs inhibit retinoic acid-induced apoptosis in Hep3B cells. AB - Retinoic acid (RA) induces apoptosis in Hep3B human hepatoma cells. 9-Cis-RA (c RA) had a similar effect as all-trans-RA (t-RA) in inducing cell death in Hep3B cells. RA-induced Hep3B-cell death was associated with inhibited expression of the hepatocyte nuclear factor 4 (HNF-4) gene. Palmitoyl-CoA ((C16:0)-CoA), the reported HNF-4 ligand, prevented RA-induced apoptosis. The effect of (C16:0)-CoA was specific, since palmitic acid and co-enzyme A had no effect in preventing RA induced apoptosis. Bovine serum albumin (BSA) also prevented RA-induced apoptosis. However, in contrast to BSA, which induced cell growth, (C16:0)-CoA alone had no effect on cell growth. Investigating the possible role of HNF-4 in apoptosis, the reported HNF-4 antagonist (C18:0)-CoA was employed, and it also prevented RA-induced apoptosis. By transient transfection, overexpression of HNF 4 did not prevent RA-induced apoptosis. The induction and prevention of apoptosis caused by RA and (C16:0)-CoA were associated, respectively with the induction and inhibition of the expression of transforming growth factor beta (TGFbeta), which is known to play a role in apoptosis. Furthermore, RA and (C16:0)-CoA can regulate AP-1, which is a key regulator of the TGFbeta gene. Our data indicate that fatty acyl-CoAs can prevent RA-induced apoptosis and that TGFbeta, rather than HNF-4, may play a role in these regulatory processes. Our data also suggest that (C16:0)-CoA and (C18:0)-CoA are not the agonist and antagonist for HNF4, respectively in the Hep3B cell system. PMID- 10799736 TI - DNA breakage by resveratrol and Cu(II): reaction mechanism and bacteriophage inactivation. AB - Resveratrol (3,4',5-trihydroxy stilbene) is a phytoalexin and a polyphenolic compound present in human dietary material such as peanuts, mulberries, grapes and red wine. It is widely considered to possess cardiovascular protective properties and has also been shown to be chemopreventive against various stages of chemically induced carcinogenesis. It has recently been shown that resveratrol induces strand breakage in DNA in the presence of copper ions. In this paper, we have shown that resveratrol catalyzes the reduction of Cu(II) to Cu(I), which is accompanied by the formation of 'oxidized product(s)' of resveratrol, which in turn also appear to catalyze the reduction of Cu(II). Strand scission by the resveratrol-Cu(II) system was found to be biologically active as assayed by bacteriophage inactivation. The results are discussed in relation to the putative chemopreventive mechanism of resveratrol. PMID- 10799737 TI - Catalytic efficiencies of allelic variants of human glutathione S-transferase Pi in the glutathione conjugation of alpha, beta-unsaturated aldehydes. AB - The catalytic efficiencies of the allelic variants of human glutathione (GSH) S transferase Pi (hGSTP1-1), which differ in their primary structures by the amino acids in positions 104 (isoleucine or valine) and/or 113 (alanine or valine), in the GSH conjugation (detoxification) of acrolein and crotonaldehyde have been determined. The k(cat)/K(m) values for hGSTP1-1 isoforms I104,A113 (IA), I104, V113 (IV), V104,A113 (VA) and V104,V113 (VV) toward acrolein were 129+/-3, 116+/ 3, 128+/-4 and 92+/-3 mM(-1) s(-1), respectively. The catalytic efficiencies of the hGSTP1-1 variants IA, IV, and VA in the GSH conjugation of acrolein were statistically significantly higher (at P=0.05) compared with the VV isoform. On the other hand, the catalytic efficiencies of the hGSTP1-1 isoforms IA, IV, VA and VV toward crotonaldehyde (16+/-2, 12+/-1, 17+/-2, and 12+/-2 mM(-1)s(-1), respectively) were not statistically significantly different from each other. Our results suggest that hGSTP1-1 polymorphism may be an important factor in differential susceptibility of individuals to the toxic effects of acrolein, which is a widely spread environmental pollutant and generated endogenously during metabolic activation of anticancer drug cyclophosphamide. PMID- 10799738 TI - Concurrent administration of fish meal and sodium nitrite does not promote renal carcinogenesis in rats after initiation with N-ethyl-N-hydroxyethylnitrosamine. AB - The modifying effects of concurrent administration of fish meal and sodium nitrite on the development of renal tumors after initiation with N-ethyl-N hydroxyethylnitrosamine (EHEN) were investigated. A total of 120 male 6-week-old Wistar rats were divided into six groups. Groups 1-3 (30 animals each) were given 1000 ppm EHEN in their drinking water for 3 weeks as an initiation treatment for renal cancer induction and thereafter fed respective diets containing 64, 32, and 8% (original concentration in the basal diet) fish meal, and simultaneously given 0.12% sodium nitrite in the drinking water for 33 weeks. Groups 4-6 (ten animals each) were similarly treated without the prior application of EHEN. At the end of the 37th experimental week, all surviving animals were autopsied and examined histopathologically for the existence of renal proliferative lesions. The incidences of dysplastic lesions, adenomas or adenocarcinomas of the kidney were not significantly different among groups 1-3. No renal proliferative lesions were found in groups 4-6. Chronic nephropathy was slightly but significantly enhanced in the 64 and 32% fish meal-treated groups as compared with group 3. Our results suggest that concurrent administration of fish meal and sodium nitrite does not affect the post-initiation phase of EHEN-induced renal carcinogenesis in the rat. PMID- 10799740 TI - Inhibition by 9alpha-fluoromedoroxyprogesterone acetate (FMPA) against mammary carcinoma induced by dimethylbenz[a]anthracene in rats and angiogenesis in the rabbit cornea - comparison with medroxyprogesterone acetate (MPA). AB - Medroxyprogesterone acetate (MPA) is currently used therapeutically in the treatment of mammary and endometrial carcinomas. In order to develop a more potent and useful drug, we synthesized the novel compound, 9alpha fluoromedoroxyprogesterone acetate (FMPA), by fluorinating MPA, and we also previously reported that FMPA displays more potent anti-angiogenic activity in the chorioallantoic membrane assay than MPA. In the present study, we investigated (1) the effects of FMPA on rat mammary carcinomas induced by dimethylbenz[a]anthracene (DMBA) to determine the anti-tumor activity, (2) the effect on angiogenesis in rabbit corneal assays, and (3) compared these results with those for MPA. FMPA inhibited the growth of mammary carcinomas in a dose dependent manner (7.5, 30 and 120 mg/kg). Almost complete involution of the carcinomas was observed at doses of 30 and 120 mg/kg. MPA also inhibited the growth of carcinomas at doses of 30 and 120 mg/kg, but no involution of carcinomas was observed even at 120 mg/kg. FMPA significantly and MPA to a lesser degree inhibited carcinogenesis at 120 mg/kg within their treatments. In rabbit corneal assays, FMPA significantly inhibited angiogenesis (IC50 value=0.085 microg/pellet). MPA also significantly inhibited angiogenesis (IC50 value=0.60 microg/pellet). From these results, we conclude that FMPA is potentially more effective in the treatment of mammary carcinomas than MPA. PMID- 10799739 TI - Intercellular communication and cell proliferation in precision-cut rat liver slices: effect of medium composition and DDT. AB - Gap junctional intercellular communication (GJIC) and cell proliferation were studied in control and 1,1'-bis(p-chlorophenyl)-2, 2,2,-trichloroethane (DDT) treated precision-cut liver slices of rat by evaluating connexin 32 (Cx32) expression and 5-bromo-2'-deoxyuridine (BrdU) incorporation. In addition, the effect of different culture media (RPMI and WME) on control and DDT influenced Cx32 expression and cell proliferation was determined. Cx32 expression in control precision-cut liver slices was maintained during 8 h of culturing, but decreased after prolonged culturing. Control cell proliferation was significantly higher when WME was used as culture medium than when RPMI was used. In slices treated with DDT Cx32 expression was decreased. In slices cultured in RPMI medium, this decrease preceded a dose-dependent increase in cell proliferation. These results show the usefulness of precision-cut liver slices in studying cellular proliferation and intercellular communication. PMID- 10799741 TI - Apoptosis and p53 are associated with effect of preoperative radiation in oral squamous cell carcinomas. AB - This study was carried out to elucidate whether apoptosis and p53 can be used to stratify oral cancer patients into groups with a favorable or unfavorable response to preoperative radiation therapy. Thirty-two patients were evaluated. The apoptosis index was 1.7+/-0. 9% in the ineffective cases, and it was significantly lower than effective cases (3.2+/-1.2%). While 14 of 16 effective cases (86.7%) did not express p53, 13 of 16 ineffective cases (81.3%) overexpressed p53. These results suggest that mutated p53 in tumors is associated with a poor response to radiation which may be related to evasion of apoptosis in oral cancer. PMID- 10799742 TI - Reduction of aflatoxin B(1) adduct biomarkers by oltipraz in the tree shrew (Tupaia belangeri chinensis). AB - The risk of liver cancer is greatest in people both infected with hepatitis B virus (HBV) and highly exposed to aflatoxin B(1) (AFB(1)). The tree shrew (Tupaia belangeri chinensis) is a unique species that can be infected with human HBV, is susceptible to AFB(1)-induced liver cancer, and shows a synergistic interaction between HBV and AFB(1) for liver cancer. In this regard, the tree shrew may be useful for evaluating experimental chemoprevention strategies relevant to high risk human populations as it mirrors the human epidemiology of liver cancer. To begin developing the model for chemoprevention study, two groups of tree shrews were fed 400 microg AFB(1)/kg b.wt. in milk daily for 4 weeks. One week prior to AFB(1) administration, one group also received oltipraz (0.5 mmol/kg, p.o.) daily for 5 weeks. At weekly intervals, 1 ml of blood and a 24-h urine sample were obtained from each animal. Aflatoxin-albumin adducts in serum were determined by a radioimmunological assay and aflatoxin-N(7)-guanine adducts in urine were measured by HPLC. Aflatoxin-albumin adducts increased rapidly in 2 weeks to plateau at 20 pmol/mg protein, and they diminished after cessation of AFB(1) exposure. Oltipraz significantly attenuated the overall burden of aflatoxin albumin adducts throughout the exposure period with a median reduction of 80%. In a single cross-sectional analysis at the end of AFB(1) dosing, oltipraz treatment decreased urinary aflatoxin-N(7)-guanine by 93%. Collectively, these results indicate that oltipraz reduces AFB(1) risk biomarkers in the tree shrew in a manner similar to that observed in rodents and humans, and establishes a rationale to evaluate cancer chemoprevention by oltipraz in human HBV-infected, AFB(1) exposed tree shrews. PMID- 10799743 TI - Altered p21(WAF1/CIP1) expression is associated with poor prognosis in extrahepatic bile duct carcinoma. AB - This study was designed to determine the clinical implications of p21(WAF1/CIP1) expression and the relationship between p21(WAF1/CIP1) expression and p53 status in extrahepatic bile duct carcinoma (EBDC). Low p21(WAF1/CIP1) expression was immunohistochemically detected in 23 (67.6%) of 34 EBDCs, moderate in six (17.7%), and high in five (14.7%). Kaplan-Meier curves showed that low and high p21(WAF1/CIP1) expressions were significantly associated with shortened disease free survival (low vs. moderate, P=0.02; high vs. moderate, P=0.01). There was no correlation between p21(WAF1/CIP1) and p53 expression. These findings suggest that altered p21(WAF1/CIP1) expression exerts an adverse influence on the prognosis of EBDC. PMID- 10799744 TI - Loss of p16(INK4) protein, alone and together with loss of retinoblastoma protein, correlate with hepatocellular carcinoma progression. AB - To investigate the role of p16(INK4) protein absence in hepatocellular carcinoma progression, we examined p16(INK4) expression immunohistochemically in 81 primary and 23 metastatic lesions of hepatocellular carcinoma, in which retinoblastoma protein status had been determined. p16(INK4) protein was absent from 44% of the total of 104 tumors. The rate of p16(INK4) absence was twice as high in metastatic lesions (74%) compared with primary lesions (36%) (P=0.001). Loss of p16(INK4) and/or retinoblastoma protein was significantly associated with decreased tumor differentiation, vascular invasion and metastasis. In conclusion, p16(INK4) protein absence, alone and together with loss of retinoblastoma protein, contributes to hepatocellular carcinoma progression. PMID- 10799745 TI - Inhibitory effect of flavonoids from citrus plants on Epstein-Barr virus activation and two-stage carcinogenesis of skin tumors. AB - To search for possible anti-tumor promoters, thirteen flavones (1-13) obtained from the peel of Citrus plants were examined for their inhibitory effects on the Epstein-Barr virus early antigen (EBV-EA) activation by a short-term in vitro assay. Of these flavones, 3,5,6,7,8,3',4'-heptamethoxyflavone (HPT) (13) exhibited significant inhibitory effects on the EBV-EA activation induced by the tumor promoter, 12-O-tetradecanoylphorbol 13-acetate (TPA). Further, compound 13 exhibited remarkable inhibitory effects on mouse skin tumor promotion in an in vivo two-stage carcinogenesis test. PMID- 10799746 TI - Expression of the c-kit protein is associated with certain subtypes of salivary gland carcinoma. AB - The c-Kit protein, a receptor type tyrosine kinase that plays an important role in the development of hematopoietic cells, melanocytes, and germ cells, is expressed in mastocytosis, gastrointestinal stromal cell tumors (GISTs), germ cell tumors, and several other tumors. Gain-of-function mutations in exon 11 and exon 17 have been shown as a mechanism of c-kit activation in some tumors. To study the role of c-kit in salivary gland carcinomas, we analyzed the c-kit protein expression in 79 carcinomas of major and minor salivary glands by immunohistochemistry. Although varying in intensity of staining, c-kit expression was identified very often in adenoid cystic carcinomas (20/25), lymphoepithelioma like carcinomas (6/6) and myoepithelial carcinomas (2/2), but not in other types of salivary gland carcinoma (0/46), P<0.00001. By DNA sequencing, genetic alteration of c-kit juxtamembrane domain (exon 11) and tyrosine kinase domain (exon 17) was not found in all the three types of salivary carcinoma that had c kit protein expression. In conclusion, c-kit protein overexpression is involved in the pathogenesis of certain types of salivary gland carcinoma, but mutation of the gene is not the mechanism of c-kit activation. PMID- 10799747 TI - Epothilones and related structures--a new class of microtubule inhibitors with potent in vivo antitumor activity. PMID- 10799748 TI - p63 and p73: old members of a new family. PMID- 10799749 TI - Multiplying functions for BRCA1 and BRCA2?. Meeting report, The Breakthrough Breast Cancer Second International Workshop on the function of BRCA1 and BRCA2, Cambridge, UK, 9-10 September 1999. PMID- 10799750 TI - Molecular horizons of cancer therapeutics: 11th Pezcoller symposium. PMID- 10799751 TI - Dominant negative effects of apolipoprotein E4 revealed in transgenic models of neurodegenerative disease. AB - Apolipoprotein E fulfills fundamental functions in lipid transport and neural tissue repair after injury.(6,8) Its three most common isoforms (E2, E3, and E4) are critical determinants of diverse human diseases, including major cardiovascular and neurodegenerative disorders.(8,14) Apolipoprotein E4 is associated with an increased risk for Alzheimer's disease(3,5) and poor clinical outcome after head injury or stroke.(11,16) The precise role of apolipoprotein E4 in these conditions remains unknown. To characterize the effects of human apolipoprotein E isoforms in vivo, we analysed transgenic Apoe knockout mice that express apolipoprotein E3 or E4 or both in the brain. Hemizygous and homozygous apolipoprotein E3 mice were protected against age-related and excitotoxin-induced neurodegeneration, whereas apolipoprotein E4 mice were not. Apolipoprotein E3/E4 bigenic mice were as susceptible to neurodegeneration as apolipoprotein E4 singly transgenic mice. At eight months of age neurodegeneration was more severe in homozygous than in hemizygous apolipoprotein E4 mice consistent with a dose effect. Thus, apolipoprotein E4 is not only less neuroprotective than apolipoprotein E3 but also acts as a dominant negative factor that interferes with the beneficial function of apolipoprotein E3. The inhibition of this apolipoprotein E4 activity may be critical for the prevention and treatment of neurodegeneration in APOE varepsilon4 carriers. PMID- 10799752 TI - Dopamine D(4) receptors in rat forebrain: unchanged with amphetamine-induced behavioral sensitization. AB - Dopamine D(2) receptors are implicated in stimulant-induced behavioral sensitization.(7,10) Studies using selective receptor antagonists also implicate the D(4) receptor, a member of the dopamine D(2)-like receptor family.(3) Accordingly, dopamine D(4) and D(2)-like receptor levels in rat forebrain were examined by computed autoradiography after repeated (+)-amphetamine treatment that induced behavioral sensitization. Receptor binding was quantified in critical brain regions including caudate-putamen, nucleus accumbens septi, medial prefrontal cortex and hippocampus. No significant differences in D(4) or D(2) like receptor levels were detected among rats sensitized to amphetamine, those exposed to amphetamine but killed before behavioral sensitization emerged or vehicle-treated controls. The findings indicate that expression of amphetamine induced behavioral sensitization is not associated with altered D(4) (or D(2)) receptor density in rat forebrain. PMID- 10799753 TI - Distance of target search of isolated rat hippocampal neuron is about 150 microm. AB - Although the survival of neuronal cells is highly dependent on neural connections with afferents or targets,(10,14,15) little is known about the survival of immature neurons that have not yet encountered the partners. Herein, using cultures of isolated hippocampal neurons of rat embryos, we have attempted to elucidate the contribution of neurite outgrowth to neuron survival and found that neurons died at a certain degree of neurite length with apoptotic characteristics in cases of no contact with other neurons. The threshold was 143.4microm, which was about five times as long as the cell body diameter. It was altered by depolarization or in the presence of basic fibroblast growth factor. Thus, neurons may be designed to kill themselves if they cannot find their targets after exploration within a particular area, the extent of which is variable due to cellular conditions. PMID- 10799754 TI - Inhibition of synaptically evoked cortical acetylcholine release by adenosine: an in vivo microdialysis study in the rat. AB - The release of cortical acetylcholine from the intracortical axonal terminals of cholinergic basal forebrain neurons is closely associated with electroencephalographic activity. One factor which may act to reduce cortical acetylcholine release and promote sleep is adenosine. Using in vivo microdialysis, we examined the effect of adenosine and selective adenosine receptor agonists and antagonists on cortical acetylcholine release evoked by electrical stimulation of the pedunculopontine tegmental nucleus in urethane anesthetized rats. All drugs were administered locally within the cortex by reverse dialysis. None of the drugs tested altered basal release of acetylcholine in the cortex. Adenosine significantly reduced evoked cortical acetylcholine efflux in a concentration-dependent manner. This was mimicked by the adenosine A(1) receptor selective agonist N(6)-cyclopentyladenosine and blocked by the selective A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). The A(2A) receptor agonist 2-[p-(2-carboxyethyl)-phenethylamino]-5'-N ethylcarboxamidoadenosi ne hydrochloride (CGS 21680) did not alter evoked cortical acetylcholine release even in the presence of DPCPX. Administered alone, neither DPCPX nor the non-selective adenosine receptor antagonist caffeine affected evoked cortical acetylcholine efflux. Simultaneous delivery of the adenosine uptake inhibitors dipyridamole and S-(4-nitrobenzyl)-6-thioinosine significantly reduced evoked cortical acetylcholine release, and this effect was blocked by the simultaneous administration of caffeine. These data indicate that activation of the A(1) adenosine receptor inhibits acetylcholine release in the cortex in vivo while the A(2A) receptor does not influence acetylcholine efflux. Such inhibition of cortical acetylcholine release by adenosine may contribute to an increased propensity to sleep during prolonged wakefulness. PMID- 10799755 TI - Heterogeneous populations of cells mediate spontaneous synchronous bursting in the developing hippocampus through a frequency-dependent mechanism. AB - Under normal conditions, hippocampal slices from newborn rats and rabbits (postnatal days 0-8) show spontaneous synchronous bursts known as giant depolarizing potentials. These bursts are recorded from CA3, CA1 and the fascia dentata in both intact slices and isolated hipocampal regions. Giant depolarizing potentials are network-driven events resulting from the synergistic activation of N-methyl-D-aspartate, alpha-amino-3-hydroxy-5-methyl-4-isoxadepropionate and GABA(A) receptors, the latter playing an excitatory role. Recently, we showed that they spontaneously emerge in an all-or-none manner after the increase of synaptic and cellular activity beyond a threshold frequency [Menendez de la Prida L. and Sanchez-Andres J. V. (1999) J. Neurophysiol. 82, 202-208]. Under this framework, background levels of spontaneous activity at individual neurons build up network synchronization 100-300ms prior to the onset of giant depolarizing potentials. However, the role of distinct cellular populations and connectivity in determining the threshold frequency has not been examined. By performing simultaneous intracellular recordings from pyramidal cells, non-pyramidal cells and interneurons, we investigated their participation in the generation of giant depolarizing potentials. Electrodes containing Neurobiotin were used to examine the cellular morphology. We found that giant depolarizing potentials were not initiated from a single pacemaker cellular group; instead, they involved recurrent cooperation among these groups, which contributed differently according to their intrinsic firing capability. In all the neurons examined, the onset of these bursts took place in an all-or-none frequency-dependent manner, both spontaneously (depending on the frequency of the excitatory postsynaptic potentials) or when triggered by extracellular stimulation. The CA3 threshold of frequency was at 12Hz in both pyramidal cells and interneurons, while in the fascia dentata it was 17Hz. The application of 6-cyano-7-nitroquinoxaline-2,3 dione increased CA3 threshold of frequency up to 50Hz, suggesting that it is determined by combined synaptic components. We examined the role of postsynaptic summation on the threshold of frequency. Heterogeneity is present among the cellular groups, pyramidal neurons from CA1 and CA3 showing less evidence of postsynaptic summation prior to giant depolarizing potentials. Cells showing stronger evidence of postsynaptic summation were more typically recorded at the hilus, the granule layer of the fascia dentata and the CA3/CA4 area. Nevertheless, for a given cell, not all the giant depolarizing potentials were preceded by summation of postsynaptic potentials. These outcomes, together with the long and variable time delays recorded between different areas, strongly suggest that giant depolarizing potentials are locally generated from different initiation sites and not from a single region. We discuss these results in view of the principles underlying hyperexcitability in hippocampal slices, i.e. the intrinsic firing properties of individual cells and the connectivity patterns. PMID- 10799756 TI - Kynurenergic manipulations influence excitatory synaptic function and excitotoxic vulnerability in the rat hippocampus in vivo. AB - Competing enzymatic mechanisms degrade the tryptophan metabolite L-kynurenine to kynurenate, an inhibitory and neuroprotective compound, and to the neurotoxins 3 hydroxykynurenine and quinolinate. Kynurenine 3-hydroxylase inhibitors such as PNU 156561 shift metabolism towards enhanced kynurenate production, and this effect may underlie the recently discovered anticonvulsant and neuroprotective efficacy of these drugs. Using electrophysiological and neurotoxicological endpoints, we now used PNU 156561 as a tool to examine the functional interplay of kynurenate, 3-hydroxykynurenine and quinolinate in the rat hippocampus in vivo. First, population spike amplitude in area CA1 and the extent of quinolinate induced excitotoxic neurodegeneration were studied in animals receiving acute or prolonged intravenous infusions of L-kynurenine, PNU 156561, (L-kynurenine+PNU 156561) or kynurenate. Only the latter two treatments, but not L-kynurenine or PNU 156561 alone, caused substantial inhibition of evoked responses in area CA1, and only prolonged (3h) infusion of (L-kynurenine+PNU 156561) or kynurenate was neuroprotective. Biochemical analyses in separate animals revealed that the levels of kynurenate attained in both blood and brain (hippocampus) were essentially identical in rats receiving extended infusions of L-kynurenine alone or (L-kynurenine+PNU 156561) (4 and 7microM, respectively, after an infusion of 90 or 180min). However, addition of the kynurenine 3-hydroxylase inhibitor resulted in a significant decrement in the formation of 3-hydroxykynurenine and quinolinate in both blood and brain. These data suggest that the ratio between kynurenate and 3-hydroxykynurenine and/or quinolinate in the brain is a critical determinant of neuronal excitability and viability. The anticonvulsant and neuroprotective potency of kynurenine 3-hydroxylase inhibitors may therefore be due to the drugs' dual action on both branches of the kynurenine pathway of tryptophan degradation. PMID- 10799757 TI - Reorganization of the morphology of hippocampal neurites and synapses after stress-induced damage correlates with behavioral improvement. AB - We recently demonstrated that stress-induced cognitive deficits in rats do not correlate with hippocampal neuronal loss. Working on the premise that subtle structural changes may however be involved, we here evaluated the effects of chronic stress on hippocampal dendrite morphology, the volume of the mossy fiber system, and number and morphology of synapses between mossy fibers and CA3 dendritic excrescences. To better understand the mechanisms by which stress exerts its structural effects, we also studied these parameters in rats given exogenous corticosterone. Further, to search for signs of structural reorganization following the termination of the stress and corticosterone treatments, we analysed groups of rats returned to treatment-free conditions. All animals were assessed for spatial learning and memory performance in the Morris water maze. Consistent with previous findings, dendritic atrophy was observed in the CA3 hippocampal region of chronically stressed and corticosterone-treated rats; in addition, we observed atrophy in granule and CA1 pyramidal cells following these treatments. Additionally, profound changes in the morphology of the mossy fiber terminals and significant loss of synapses were detected in both conditions. These alterations were partially reversible following rehabilitation from stress or corticosterone treatments. The fine structural changes, which resulted from prolonged hypercortisolism, were accompanied by impairments in spatial learning and memory; the latter were undetectable following rehabilitation. We conclude that there is an intimate relationship between corticosteroid levels, hippocampal neuritic structure and hippocampal-dependent learning and memory. PMID- 10799758 TI - Evidence for striatal dopaminergic overactivity in paroxysmal dystonia indicated by microinjections in a genetic rodent model. AB - Mutant dystonic hamsters (dt(sz)), a model of primary paroxysmal dystonia, display attacks of generalized dystonia in response to mild stress in an age dependent manner. Recent studies in dystonic hamsters have revealed decreased densities of dopamine D(1) and D(2) in the dorsal striatum. This finding has been interpreted as a down-regulation in response to enhanced dopamine release because systemic treatments with neuroleptics reduced the severity of dystonia while levodopa exerted prodystonic effects. Therefore, in the present study we investigated the effects of amphetamine as well as of selective D(1) or D(2) receptor agonists and antagonists on the severity of dystonia after systemic administrations and after microinjections into the dorsal striatum. Amphetamine and the dopamine D(2) agonist quinpirole increased the severity of dystonia after systemic and striatal injections, while the dopamine D(1) agonist SKF 38393 exerted only moderate prodystonic effects after systemic administration of a high dose but not after striatal injections. These results suggest that a predominant overstimulation of D(2) receptors is pathogenetically involved in the dystonic syndrome. Combined systemic or striatal administrations of the D(1) and D(2) receptor agonists did not reveal synergistic prodystonic effects at the examined doses. The selective D(1) antagonist SCH 23390 as well as the D(2) antagonist raclopride tended to decrease the severity of dystonia after systemic administration but failed to exert significant effects after striatal injection. The coadministration of ineffective doses of the antagonists SCH 23390 and raclopride, however, exerted an enormous antidystonic efficacy after both systemic and striatal injections. Since striatal injections of compounds which enhance dopaminergic activity aggravated dystonia, while coinjections of dopamine D1 and D2 receptor antagonists reduced the severity of dystonia, the present findings clearly support the hypothesis that striatal dopaminergic overactivity plays a crucial role for the manifestation of dystonic attacks in the hamster model of paroxysmal dystonia. PMID- 10799759 TI - Enhanced vulnerability to oxidative stress by alpha-synuclein mutations and C terminal truncation. AB - alpha-Synuclein is a key component of Lewy bodies found in the brains of patients with Parkinson's disease and two point mutations in this protein, Ala53Thr and Ala30Pro, are associated with rare familial forms of the disease. Several lines of evidence suggest the involvement of oxidative stress in the pathogenesis of nigral neuronal death in Parkinson's disease. In the present work we studied the effects of changes in the alpha-synuclein sequence on the susceptibility of cells to reactive oxygen species. Human dopaminergic neuroblastoma SH-SY5Y cells were stably transduced with various isoforms of alpha-synuclein and their survival following exposure to hydrogen peroxide or to the dopaminergic neurotoxin MPP(+) was assessed. Cells expressing the two point mutant isoforms of alpha-synuclein were significantly more vulnerable to oxidative stress, with the Ala53Thr engineered cells faring the worst. In addition, cells expressing C-terminally truncated alpha-synuclein, particularly the 1-120 residue protein, were more susceptible than control beta-galactosidase engineered cells. The present experiments indicate that point mutations and C-terminal truncation of alpha synuclein exaggerate the susceptibility of dopaminergic cells to oxidative damage. Thus, these observations provide a pathogenetic link between alpha synuclein aberrations and a putative cell death mechanism in Parkinson's disease. PMID- 10799760 TI - Naloxone prevents microglia-induced degeneration of dopaminergic substantia nigra neurons in adult rats. AB - Resident microglia are involved in immune responses of the central nervous system and may contribute to neuronal degeneration and death. Here, we tested in adult rats whether injection of bacterial lipopolysaccharide (which causes inflammation and microglial activation) just above the substantia nigra, results in the death of dopaminergic substantia nigra pars compacta neurons. Two weeks after lipopolysaccharide injection, microglial activation was evident throughout the nigra and the number of retrogradely-labeled substantia nigra neurons was reduced to 66% of normal. This suggests that inflammation and/or microglial activation can lead to neuronal cell death in a well-defined adult animal model. The opioid receptor antagonist naloxone reportedly reduces release of cytotoxic substances from microglia and protects cortical neurons in vitro. Here, a continuous two week infusion of naloxone at a micromolar concentration close to the substantia nigra, prevented most of the neuronal death caused by lipopolysaccharide, i.e. 85% of the neurons survived. In addition, with systemic (subcutaneous) infusion of 0. 1mg/d naloxone, 94% of the neurons survived. Naloxone infusions did not obviously affect the morphological signs of microglial activation, suggesting that naloxone reduces the release of microglial-derived cytotoxic substances. Alternatively, microglia might not cause the neuronal loss, or naloxone might act by blocking opioid receptors on (dopaminergic or GABAergic) neurons.Thus, local inflammation induces and the opioid antagonist naloxone prevents the death of dopaminergic substantia nigra neurons in adult rats. This may be relevant to the understanding of the pathology and treatment of Parkinson's disease, where these neurons degenerate. PMID- 10799761 TI - Neurotensin regulates intracellular calcium in ventral tegmental area astrocytes: evidence for the involvement of multiple receptors. AB - Recent evidence suggests that some types of neurotensin receptors may be expressed by astrocytes. In order to explore the function of neurotensin receptors in astrocytes, the effect of a neurotensin receptor agonist, neurotensin(8-13), on intracellular Ca(2+) dynamics in mixed neuronal/glial cultures prepared from rat ventral tegmental area was examined. It was found that neurotensin(8-13) induces a long-lasting rise in intracellular Ca(2+) concentration in a subset of glial fibrilary acidic protein-positive glial cells. This response displays extensive desensitization and appears to implicate both intracellular and extracellular Ca(2+) sources. In the absence of extracellular Ca(2+), neurotensin(8-13) evokes only a short-lasting rise in intracellular Ca(2+). The neurotensin-evoked intracellular Ca(2+) accumulation is blocked by the phospholipase C inhibitor U73122 and by thapsigargin, suggesting that it is initiated by release of Ca(2+) from an inositol triphosphate-dependent store. The Ca(2+)-mobilizing action of neurotensin(8-13) in astrocytes is dependent on at least two receptors, because the response is blocked in part only by SR48692, a type 1 neurotensin receptor antagonist, and is blocked completely by SR142948A, a novel neurotensin receptor antagonist. The finding that the type 2 neurotensin receptor agonist levocabastine fails to mimic or alter the effects of neurotensin(8-13) on intracellular Ca(2+) makes it unlikely that the type 2 neurotensin receptor is involved. In summary, these results show that functional neurotensin receptors are present in cultured ventral tegmental area astrocytes and that their activation induces a highly desensitizing rise in intracellular Ca(2+). The pharmacological profile of this response suggests that a type 1 neurotensin receptor is involved but that another, possibly novel, non-type 2 neurotensin receptor is also implicated. If present in vivo, such signalling could be involved in some of the physiological actions of neurotensin. PMID- 10799762 TI - Brain-derived neurotrophic factor modulates dendritic morphology of cerebellar basket and stellate cells: an in vitro study. AB - The dendrites of cerebellar basket/stellate cells show a highly stereotyped orientation relative to granule cell axons (parallel fibers) and Purkinje cell dendrites. This specific morphology is acquired during the early postnatal phase of cerebellar development, when basket/stellate cells become synaptically integrated with Purkinje neurons and granule cells. In the present study, we used primary cerebellar cultures to test how the spatial arrangement of granule cell axons affects basket/stellate cell dendritic morphology. In addition, we sought to determine whether active signals as might be provided by granule cells, i.e. synaptic input and the neurotrophin, brain-derived neurotrophic factor, affect basket/stellate cell development. Our results confirm the critical role of parallel fiber orientation for basket/stellate dendritic morphogenesis. Moreover, we found that both electrical activity and brain-derived neurotrophic factor increased basket and stellate cell dendritic arborization. Together with previously published findings, our data led to the conclusion that both structural cues and active interneuronal signaling collaborate to bring about the precise morphogenesis of cerebellar basket/stellate cells. The distinct responses of various cerebellar phenotypes towards the morphogenetic effects of brain derived neurotrophic factor suggest that this neurotrophin, within the developing cerebellum, enhances synaptic connectivity by concerting the formation of appropriate pre- and postsynaptic structures. PMID- 10799763 TI - Effects of persistent nociception on periaqueductal gray glycine release. AB - Glycine is a candidate nociception inhibitory transmitter in specific brain regions, like for example the spinal cord, the thalamic nuclei and the periaqueductal gray matter. However, quantitative changes in glycine released in these brain regions during peripheral inflammation episodes have not been characterized in awake animals. To address this issue, an in vivo microdialysis study was carried out in freely moving rats in order to analyse periaqueductal gray matter extracellular glycine concentration following unilateral formalin injection into the dorsal skin of the right hind-paw. The extracellular concentration of glutamine was also evaluated in order to analyse whether or not a non-neurotransmitter amino acid was equally modified. Intra-periaqueductal gray matter tetrodotoxin perfusion reduced extracellular glycine concentration (-44+/ 5%), but did not change the glutamine dialysate values. Peripheral injection of formalin reduced the glycine release during the early phase (-62+/-8%) and the late phase (-36+/-6%) of hyperalgesia, although not during the analgesic period. Perfusion with naloxone (300microM) neither prevented the formalin-induced decreases in extacellular glycine concentration, nor modified the perfusate basal values of glycine and glutamine. These results show that, contrary to what has been recognized on the interactive role of opioids and GABA into the periaqueductal gray matter (i.e. opioid disinhibition), endogenous opioids seem not to modulate the activity of glycinergic neurons in the same midbrain area. In the light of these preliminary data, it is reasonable to suppose that GABA and glycine are probably not co-released at the level of periaqueductal gray matter of the rat. PMID- 10799764 TI - Lemniscal recurrent and transcortical influences on cuneate neurons. AB - Intracellular recordings were obtained from cuneate neurons of chloralose anesthetized, paralysed cats to study the synaptic responses induced by electrical stimulation of the contralateral medial lemniscus. From a total of 178 cells sampled, 109 were antidromically fired from the medial lemniscus, 82 of which showed spontaneous bursting activity. In contrast, the great majority (58/69) of the non-lemniscal neurons presented spontaneous single spike activity. Medial lemniscus stimulation induced recurrent excitation and inhibition on cuneolemniscal and non-lemniscal cells. Some non-lemniscal neurons were activated by somatosensory cortex and inhibited by motor cortex stimulation. Some other non lemniscal cells that did not respond to medial lemniscus stimulation in control conditions were transcortically affected by stimulating the medial lemniscus after inducing paroxysmal activity in the sensorimotor cortex. These findings indicate that different sites in the sensorimotor cortex can differentially influence the sensory transmission through the cuneate, and that the distinct available corticocuneate routes are selected within the cerebral cortex. From a total of 92 cells tested, the initial effect induced by low-frequency stimulation of the sensorimotor cortex was inhibition on most of the cuneolemniscal neurons (32/52) and excitation on the majority of the non-lemniscal cells (25/40). The fact that a substantial proportion of cuneolemniscal and non-lemniscal cells was excited and inhibited, respectively, suggests that the cerebral cortex may potentiate certain inputs by exciting and disinhibiting selected groups of cuneolemniscal cells. Finally, evidence is presented demonstrating that the tendency of the cuneolemniscal neurons to fire in high-frequency spike bursts is due to different mechanisms, including excitatory synaptic potentials, recurrent activation through lemniscal axonal collaterals, and via the lemnisco-thalamo cortico-cuneate loop.A corticocuneate network circuit to explain the results is proposed. PMID- 10799765 TI - Spinal lamina I neurons that express neurokinin 1 receptors: morphological analysis. AB - The morphology of neurons in lamina I of the dorsal horn of the lumbar spinal cord which express neurokinin 1 receptors in the rat has been investigated. On the basis of soma and dendritic measurements, these neurons form two populations. One group consists of large neurons that stain intensely for the neurokinin 1 receptor with the immunochemical methods employed. They have a large soma, typically giving rise to between three and five thick principal dendrites. The dendritic tree, however, is relatively sparse, with the principal dendrites giving rise to small numbers of second- and third-order branches. All these dendrites are almost spine free. The dendritic tree spreads extensively in the rostrocaudal (approximately 550microm) and mediolateral (approximately 30microm) orientations, with few ventrally directed branches. These cells give rise to a single axon from their soma or a principal dendrite that generates a few local branches and also ramifies sparsely in deeper laminae (II-IV). The details of axonal morphology were established from intracellularly labelled material. Ultrastructural analysis of the synaptic input to these neurons reveals that they receive synapses with both clear round, flattened and dense-core vesicles; however, they do not form components of glomerular synapses. The second neuron type stains less intensely and typically has a small fusiform soma, giving rise to dendrites at its rostral and caudal poles. The dendritic tree is long in the rostrocaudal orientation (approximately 350microm), but restricted mediolaterally (approximately 40microm). The primary dendrites of these neurons bifurcate and soon give rise to third-order branches that are spiny. No pattern of organization could be detected for the distribution of either neuron type. These observations are discussed in the light of other recent studies indicating a central role for lamina I neurons expressing neurokinin 1 in the perception of severe pain. PMID- 10799766 TI - Mechanosensitive chloride channels on the growth cones of cultured rat dorsal root ganglion neurons. AB - We developed an experimental system to investigate mechanosensitivity of a single neuron, using cultured rat dorsal root ganglion cells. Highly precise mechanical stimulation was applied to various sites of the cells, using a piezo-driven glass microcapillary whose movement was computer-controlled, while whole-cell patch clamp recordings were made from the cell bodies. When the growth cones and lamillipodia from the cell soma were mechanically stimulated, inward currents were recorded at the holding potential of -60mV. Filopodia were most sensitive to mechanical stimulation. However, when neurites or soma of dorsal root ganglion cells were stimulated in the same way, electrical responses were hardly recorded. Two types of currents varying in time-course were observed: fast type of 100 200ms and slow type of several seconds in duration. When the membrane potential was held at around 0mV, both types of currents were almost abolished or even reversed, and the reversal potential was estimated to be about -2. 2mV. Replacement of extracellular sodium by tetraethylammonium did not significantly change the reversal potential. In the low-chloride solution ([Cl(-)](o)=11.7mM), the reversal potential was about +60mV, as expected from the Nernst equation for chloride. These inward currents were almost completely inhibited by extracellular application of chloride channel blocker, 5-nitro-2-(3-phenylpropylamino) benzoic acid (100microM). These results indicate that the inward currents are due to activation of mechanosensitive chloride channels, preferentially located on the growth cones of cultured dorsal root ganglion neurons. PMID- 10799767 TI - Reducing transmitter release from nerve terminals influences motoneuron survival in developing rats. AB - Motoneurons in neonatal rats die following injury to the peripheral nerve. However, this vulnerability to nerve injury declines rapidly so that nerve injury at five days of age results in little if any motoneuron death. We have proposed that the role of the target during this critical period of development is to up regulate the release of transmitter from developing motor nerve terminals. Here we show that reducing the release of acetylcholine from nerve terminals in neonatal rats can affect motoneuron maturation and survival. The soleus muscle in neonatal rats was treated with either magnesium or hemicholinium, and the number of motoneurons that survived was established 10 weeks later by retrograde labelling. Following treatment with magnesium, only 58.1% (+/-10.4 S.E.M., n=5) of the motoneurons in the soleus motor pool survived, although hemicholinium had no effect on motoneuron survival. However, those motoneurons that survived following treatment with either magnesium or hemicholinium did not develop normally since they remained susceptible to axotomy-induced cell death for longer than normal. In adult animals in which the sciatic nerve was crushed at five days of age following prior treatment with either magnesium or hemicholinium, only 27.6% (+/-6.2 S.E.M., n=5) and 44% (+/-6.1 S.E.M., n=4) of motoneurons in the sciatic motor pool survived, respectively, although no motoneurons died following injury alone or when injury was preceded by treatment with control implants containing NaCl. These results indicate that the release of acetylcholine from motor nerve terminals plays an important role in the development and survival of motoneurons. PMID- 10799768 TI - Mitochondrial porin required for ischemia-induced mitochondrial dysfunction and neuronal damage. AB - The precise molecular events of mitochondrial dysfunction, one of the last steps that irreversibly determines cellular degeneration and death, remain unknown. We introduce a novel strategy to isolate and assess the molecular mechanisms underlying mitochondrial dysfunction. Using an in vitro ischemia model, we obtained evidence for prolonged mitochondrial depolarization in rat organotypic hippocampal brain slices during reperfusion. Then, mitochondria were isolated from brain slices and mitochondrial proteins were purified on a cyclosporin-A affinity column. Cyclosporin-A is the most potent inhibitor of mitochondrial dysfunction, in particular the mitochondrial permeability transition, and therefore we hypothesized that it may interact with proteins involved in the permeability transition after mitochondria were subjected to manipulations that promote this event. Mitochondrial porin was reproducibly eluted from the affinity column using proteins from ischemic brain mitochondria, or from mitochondria exposed to oxidative stress that were used as a positive control. Anti-porin antibodies prevented mitochondrial depolarization and electrophysiological deterioration of hippocampal neurons during hypoxia-reperfusion, as measured by simultaneous fluorescence imaging and whole-cell recordings. These observations provide biochemical and functional evidence that porin is directly involved in mitochondrial dysfunction and neuronal impairment during ischemia-reperfusion, and indicate that porin could be a novel therapeutic target to prevent cellular degeneration. PMID- 10799769 TI - Astrocytes in sensory circumventricular organs of the rat brain express functional binding sites for endothelin. AB - Sensory circumventricular organs bordering the anterior third cerebral ventricle, the subfornical organ and the organum vasculosum laminae terminalis, lack blood brain barrier characteristics and are therefore accessible to circulating peptides like endothelins. Astrocytes of the rat subfornical organ and organum vasculosum laminae terminalis additionally showed immunocytochemical localization of endothelin-1/endothelin-3-like peptides, possibly acting as circumventricular organ-intrinsic modulators. Employing [125I]endothelin-1 as radioligand, quantitative autoradiography demonstrated specific binding sites throughout the rat organum vasculosum laminae terminalis and subfornical organ, and competitive displacement studies revealed expression of both ET(A) and ET(B) receptor subtypes for either circumventricular organ. ET(B) receptor binding prevailed for the whole brain and ET(A) receptors could be labelled in the peripheral vascular system. To characterize endothelin-specific receptors in astrocytes of both circumventricular organs, alterations in the intracellular calcium concentration due to endothelin-1/endothelin-3 stimulation were studied in primary culture of subfornical organ and organum vasculosum laminae terminalis cells obtained from early postnatal rat pups. Endothelin-1 and endothelin-3 induced Ca(2+) transients in 9-13% of either subfornical organ or organum vasculosum laminae terminalis astrocytes, respectively, and some glial cells (subfornical organ: 2%, organum vasculosum laminae terminalis: 5%) responded to both endothelin analogues. The antagonistic action of BQ123 specific for ET(A) receptors (74% of all astrocytes tested), and the pronounced responsiveness to the ET(B) receptor agonist [4Ala]ET 1 (subfornical organ: 27%, organum vasculosum laminae terminalis: 35%) demonstrated glial expression of both endothelin receptor subtypes. Agonist induced elevations in the intracellular calcium concentration proved to be independent of extracellular Ca(2+). In summary, the results indicate that endothelin(s) interact(s) with circumventricular organ astrocytes. Competitive receptor binding techniques using brain tissue sections as well as a fura-2 loaded primary cell culture system of the subfornical organ and organum vasculosum laminae terminalis demonstrate glial expression of functional ET(A) and ET(B) receptors, with calcium as intracellular messenger emerging primarily from intracellular stores. Endothelin(s) of both circulating and circumventricular organ-intrinsic origin may afferently transfer information important for cardiovascular homeostasis to circumventricular organs serving as "windows to the brain". PMID- 10799770 TI - Role of nitric oxide in the ethylcholine aziridinium model of delayed apoptotic neurodegeneration in vivo and in vitro. AB - The involvement of nitric oxide in neurodegenerative processes still remains incompletely characterized. Although nitric oxide has been reported to be an important mediator in neuronal degeneration in different models of cell death involving NMDA-receptor activation, increasing evidence for protective mechanisms has been obtained. In this study the role of nitric oxide was investigated in a model of NMDA-independent, delayed apoptotic cell death, induced by the neurotoxin ethylcholine aziridinium ethylcholine aziridinium both in vivo and in vitro. For the in vivo evaluation rats received bilateral intracerebroventricular injections of ethylcholine aziridinium (2nmol/ventricle) or vehicle. In the hippocampus a transient decrease in nitric oxide synthase activity occurred, reaching its lowest levels three days after ethylcholine aziridinium treatment (51.7+/-9.8% of controls). The decrease coincided with the maximal reduction in choline acetyltransferase activity as marker for the extent of cholinergic lesion. The effect of pharmacological inhibition of nitric oxide synthase was tested by application of various nitric oxide synthase inhibitors with different selectivity for the nitric oxide synthase-isoforms. Unspecific nitric oxide synthase inhibition resulted in a significant potentiation of the loss of choline acetyltransferase activity in the hippocampus measured seven days after ethylcholine aziridinium application, whereas the specific inhibition of neuronal or inducible nitric oxide synthase was ineffective. These pharmacological data are suggestive for a neuroprotective role of nitric oxide generated by endothelial nitric oxide synthase. In vitro experiments were performed using serum-free primary neuronal cell cultures from hippocampus, cortex and septum of E15-17 Wistar rat embryos. Ethylcholine aziridinium-application in a range of 5 80microM resulted in delayed apoptotic neurodegeneration with a maximum after three days as confirmed by morphological criteria, life-death assays and DNA laddering. Nitric oxide synthase activity in harvested cells decreased in a dose- and time-dependent manner. Nitric oxide production as determined by measurement of the accumulated metabolite nitrite in the medium was equally low in controls and in ethylcholine aziridinium treated cells (range 0.77-1.86microM nitrite). An expression of inducible nitric oxide synthase messenger RNA could not be detected by semiquantitative RT-PCR 13h after ethylcholine aziridinium application. The present data indicate that in a model of delayed apoptotic neurodegeneration as induced by ethylcholine aziridinium neuronal cell death in vitro and in vivo is independent of the cytotoxic potential of nitric oxide. This is confirmed by a decrease in nitric oxide synthase activity, absence of nitric oxide production and absence of inducible nitric oxide synthase expression. In contrast, evidence for a neuroprotective role of nitric oxide was obtained in vivo as indicated by the exaggeration of the cholinergic lesion after unspecific nitric oxide synthase inhibition by N-nitro-L-arginine methylester. PMID- 10799771 TI - Regionally selective changes in brain lysosomes occur in the transition from young adulthood to middle age in rats. AB - The possibility that brain aging in rats exhibits regional variations of the type found in humans was studied using lysosomal chemistry as a marker. Age-related (two vs 12months; male Sprague-Dawley) differences in cathepsin D immunostaining were pronounced in the superficial layers of entorhinal cortex and in hippocampal field CA1, but not in neocortex and field CA3. Three changes were recorded: an increase in the intraneuronal area occupied by labeled lysosomes; clumping of immunopositive material within neurons; more intense cytoplasmic staining. Western blot analyses indicated that the increases involved the active forms of cathepsin D rather than their proenzyme. Shrinkage of cathepsin-D-positive neuronal cell bodies was observed in entorhinal cortex but not in neocortical sampling zones. Age-related lysosomal changes as seen with cathepsin B immunocytochemistry were considerably more subtle than those obtained with cathepsin D antibodies. In contrast, a set of glial and/or vascular elements located in a distal dendritic field of the middle-aged hippocampus was much more immunoreactive for cathepsin B than cathepsin D. The areas exhibiting sizeable changes in the present study are reported to be particularly vulnerable to aging in humans. The results thus suggest that aspects of brain aging common to mammals help shape neurosenescence patterns in humans. PMID- 10799772 TI - Dietary protein alters age-induced change in hypothalamic GABA and immune response. AB - The effect of dietary protein on hypothalamic GABAergic activity and immune response of rats in relation to age was studied. The age-induced (due to increase of age from three to 18 months) decrease in hypothalamic GABAergic activity and immune response were potentiated with the supplementation of protein rich diet under both short- and long-term conditions. Long-term consumption of protein-poor diet, in contrast, produced activation of hypothalamic GABAergic activity with an immunopotentiation with the increase of age from three to 18 months; whereas, short-term supplementation of low protein diet did not show any effect. The results of the present study may indicate that the activation or inhibition of hypothalamic GABAergic activity by immunopotentiation or immunosuppression during aging depends on the variation of the amount of dietary protein as well as the duration of its supplementation. PMID- 10799773 TI - Preface PMID- 10799774 TI - A review of avian influenza in different bird species. AB - Only type A influenza viruses are known to cause natural infections in birds, but viruses of all 15 haemagglutinin and all nine neuraminidase influenza A subtypes in the majority of possible combinations have been isolated from avian species. Influenza A viruses infecting poultry can be divided into two distinct groups on the basis of their ability to cause disease. The very virulent viruses cause highly pathogenic avian influenza (HPAI), in which mortality may be as high as 100%. These viruses have been restricted to subtypes H5 and H7, although not all viruses of these subtypes cause HPAI. All other viruses cause a much milder, primarily respiratory disease, which may be exacerbated by other infections or environmental conditions. Since 1959, primary outbreaks of HPAI in poultry have been reported 17 times (eight since 1990), five in turkeys and 12 in chickens. HPAI viruses are rarely isolated from wild birds, but extremely high isolation rates of viruses of low virulence for poultry have been recorded in surveillance studies, giving overall figures of about 15% for ducks and geese and around 2% for all other species. Influenza viruses have been shown to affect all types of domestic or captive birds in all areas of the world, but the frequency with which primary infections occur in any type of bird depends on the degree of contact there is with feral birds. Secondary spread is usually associated with human involvement, probably by transferring infective faeces from infected to susceptible birds. PMID- 10799775 TI - Evolution of avian influenza viruses. AB - Although influenza viruses can infect a wide variety of birds and mammals, the natural host of the virus is wild waterfowl, shorebirds, and gulls. When other species of animals, including chickens, turkeys, swine, horses, and humans, are infected with influenza viruses, they are considered aberrant hosts. The distinction between the normal and aberrant host is important when describing virus evolution in the different host groups. The evolutionary rate of influenza virus in the natural host reservoirs is believed to be slow, while in mammals the rate is much higher. The higher rate of evolution in mammals is thought to be a result of selective pressure on the virus to adapt to an aberrant host species. Chickens and turkey influenza virus isolates have previously and incorrectly been lumped together with wild waterfowl, gull, and shorebird influenza viruses when determining rates of evolutionary change. To determine mutational and evolutionary rates of a virus in any host species, two primary assumptions must be met: first, all isolates included in the analysis must have descended from a single introduction of the virus, and second, the outbreak must continue long enough to determine a trend. For poultry, three recent outbreaks of avian influenza meet these criteria, and the sequences of the hemagglutinin and nonstructural genes were compared. Sequences from all three outbreaks were compared to an avian influenza virus consensus sequence, which at the amino acid level is highly conserved for all the internal viral proteins. The consensus sequence also provides a common point of origin to compare all influenza viruses. The evolutionary rates determined for all three outbreaks were similar to what is observed in mammals, providing strong evidence of adaptation of influenza to the new host species, chickens and turkeys. PMID- 10799776 TI - The epidemiology and evolution of influenza viruses in pigs. AB - Pigs serve as major reservoirs of H1N1 and H3N2 influenza viruses which are endemic in pig populations world-wide and are responsible for one of the most prevalent respiratory diseases in pigs. The maintenance of these viruses in pigs and the frequent exchange of viruses between pigs and other species is facilitated directly by swine husbandry practices, which provide for a continual supply of susceptible pigs and regular contact with other species, particularly humans. The pig has been a contender for the role of intermediate host for reassortment of influenza A viruses of avian and human origin since it is the only domesticated mammalian species which is reared in abundance and is susceptible to, and allows productive replication, of avian and human influenza viruses. This can lead to the generation of new strains of influenza, some of which may be transmitted to other species including humans. This concept is supported by the detection of human-avian reassortant viruses in European pigs with some evidence for subsequent transmission to the human population. Following interspecies transmission to pigs, some influenza viruses may be extremely unstable genetically, giving rise to variants which could be conducive to the species barrier being breached a second time. Eventually, a stable lineage derived from the dominant variant may become established in pigs. Genetic drift occurs particularly in the genes encoding the external glycoproteins, but does not usually result in the same antigenic variability that occurs in the prevailing strains in the human population. Adaptation of a 'newly' transmitted influenza virus to pigs can take many years. Both human H3N2 and avian H1N1 were detected in pigs many years before they acquired the ability to spread rapidly and become associated with disease epidemics in pigs. PMID- 10799777 TI - Emergence of H3N2 reassortant influenza A viruses in North American pigs. AB - In late summer through early winter of 1998, there were several outbreaks of respiratory disease in the swine herds of North Carolina, Texas, Minnesota and Iowa. Four viral isolates from outbreaks in different states were analyzed, both antigenically and genetically. All of the isolates were identified as H3N2 influenza viruses with antigenic profiles similar to those of recent human H3 strains. Genotyping and phylogenetic analysis demonstrated that the four swine viruses had emerged through two different pathways. The North Carolina isolate is the product of genetic reassortment between human and swine influenza viruses, while the others arose from reassortment of human, swine and avian viral genes. The hemagglutinin genes of the four isolates were all derived from the human H3N2 virus circulating in 1995. It remains to be determined if either of these recently emerged viruses will become established in the pigs in North America and whether they will become an economic burden. PMID- 10799779 TI - Host-range barrier of influenza A viruses. AB - Ample evidence suggests that all influenza viruses in mammals were probably derived from those in wild waterfowl at some time. In addition to those already established in mammals, the viruses have been transmitted to both mammals and to poultry from wild waterfowl and caused outbreaks in recent years. Experimentally, however, the viruses from one species of animals do not grow efficiently in other species. For example, human influenza viruses do not replicate in ducks or in horses, indicating their host range restriction. This paper reviews current knowledge on the host-range restriction of influenza viruses, focusing on the role of the hemagglutinin (HA). PMID- 10799778 TI - Evidence for evolutionary stasis and genetic drift by genetic analysis of two equine influenza H3 viruses isolated in France. AB - The amino acid sequences of the HA(1) portion of the haemagglutinin of two equine A(H3N8) influenza viruses isolated in France in 1993 and 1998 were analysed to determine their evolutionary relationship with 51 other HA(1) amino acid sequences available in databanks. Our data show that the French strain isolated in 1993 belongs to a group of phylogenetically related viruses branched on the main trunk, illustrating the main lineage of evolution of the equine-2 H3 sequences before its split into two distinct lineages in the late 1980s. By contrast, the 1998 French isolate appears to belong to the more recent 'Eurasian' lineage. These data suggest that equine-2 strains antigenically related to old prototype viruses may cocirculate with the more recent 'Eurasian' and 'American' lineages. In conclusion, it may be necessary to include both strains representative of recent equine influenza variants and an older prototype strain in the current equine influenza vaccines. PMID- 10799780 TI - Structural features of the avian influenza virus hemagglutinin that influence virulence. AB - Analysis of the structure of the avian influenza (AI) virus hemagglutinin (HA) gene and protein has yielded a wealth of information on the virulence mechanisms of influenza viruses. The AI hemagglutinin appears to be unique in its capacity to accept basic amino acids at its proteolytic cleavage site (PCS). The association of multiple basic (MB) amino acids, HA cleavage, tissue spread and virulence by AI strains first proposed in the late 1970s and early 1980s [Klenk, H.D., Rott, R., Orlich, M., 1977. J. Gen. Virol. 36, 151-161; Bosch, F.X., Garten, W., Klenk, H.D., Rott, R., 1981. Virology 113, 725-735] has held fast for two decades now. While other structural characteristics and other genes can certainly influence virulence, the presence of MB amino acids at the PCS has provided a hallmark structural feature which justifies continuing sequence analysis of emerging field isolates of AI strains. In addition to this structural feature, the distal tip of the HA is prone to appearance and disappearance of glycosylation sites, some of which have been associated with virulence. The recent outbreaks of highly pathogenic AI in Mexico, Australia, Pakistan, Hong Kong and in the ongoing outbreak of moderately pathogenic H7 avian influenza in the northeast US have all provided new and useful information regarding the role of HA RNA and protein structure in both virulence and host adaptation. We have previously noted that stable RNA secondary structure near the PCS is related to the acquisition of virulence and have proposed that the secondary structure may promote the insertion of basic amino acids. In this report we evaluate the phylogenetic relationships for three recent isolates of highly pathogenic avian influenza viruses and the possible virulence factors associated with their primary and secondary structure. PMID- 10799781 TI - The influenza A virus M1 protein interacts with the cellular receptor of activated C kinase (RACK) 1 and can be phosphorylated by protein kinase C. AB - The M1 protein of influenza A virus has multiple regulatory functions during the infectious cycle, which include mediation of nuclear export of viral ribonucleoproteins, inhibition of viral transcription and a crucial role in virus assembly and budding. The only known modification of the M1 protein is by phosphorylation through yet-to-be-identified kinases. We postulated that at least some of the M1 functions are exerted or regulated through interactions with cellular components. In a screen for such cellular mediators, the protein receptor of the activated C-kinase (RACK 1) was identified by its interaction with the viral M1 protein in the yeast two hybrid system. The physical M1-RACK 1 interaction was confirmed in glutathione-S-transferase-based coprecipitation assays for the diverged M1 proteins of avian, swine and human influenza A virus strains. This conservation suggests that the M1-RACK 1 interaction is of general importance during influenza A virus infections. RACK 1 has previously been identified to specifically bind the activated form of protein kinase C (PKC) and is assumed to anchor the kinase at membranes in the vicinity of its substrates. Since the M1 protein becomes phosphorylated during influenza virus infection, we examined if PKC could catalyze the phosphate transfer. We demonstrate that virion derived and recombinant M1 protein can indeed be efficiently phosphorylated by purified PKC. Moreover, in cell extracts, we detected M1 phosphorylation activity that was strongly reduced in the presence of the PKC-specific inhibitor compound GF109203X. These data suggest that PKC is the main M1-phosphorylating activity in the cell. Since both, the M1 protein and PKC have been shown to interact with RACK 1, we suggest that the M1-RACK 1 interaction is involved in M1 phosphorylation. PMID- 10799782 TI - Studies on influenza viruses H10N4 and H10N7 of avian origin in mink. AB - An influenza A virus, A/mink/Sweden/84 (H10N4), was isolated from farmed mink during an outbreak of respiratory disease, histopathologically characterised by severe interstitial pneumonia. The virus was shown to be of recent avian origin and closely related to concomitantly circulating avian influenza virus. Serological investigations were used to link the isolated virus to the herds involved in the disease outbreak. Experimental infection of adult mink with the virus isolate from the disease outbreak reproduced the disease signs and pathological lesions observed in the field cases. The mink influenza virus also induced an antibody response and spread between mink by contact. The same pathogenesis in mink was observed for two avian influenza viruses of the H10N4 subtype, circulating in the avian population. When mink were infected with the prototype avian H10 influenza virus, A/chicken/Germany/N/49, H10N7, the animals responded with antibody production and mild pulmonary lesions but neither disease signs nor contact infections were observed. Detailed studies, including demonstration of viral antigen in situ by immunohistochemistry, of the sequential development of pathological lesions in the mink airways after aerosol exposure to H10N4 or H10N7 revealed that the infections progress very similarly during the first 24h, but are distinctly different at later stages. The conclusion drawn is that A/mink/Sweden/84, but not A/chicken/Germany/N/49, produces a multiple-cycle replication in mink airways. Since the viral distribution and pathological lesions are very similar during the initial stages of infection we suggest that the two viruses differ in their abilities to replicate and spread within the mink tissues, but that their capacities for viral adherence and entry into mink epithelial cells are comparable. PMID- 10799783 TI - Cytokines in the pathogenesis of influenza. AB - Uncomplicated influenza in humans, horses or swine is characterized by massive virus replication in respiratory epithelial cells, inflammation and an abrupt onset of general and respiratory disease. There is now growing evidence that the so-called early cytokines produced at the site of infection mediate many of the clinical and pathological manifestations. Among these cytokines are interferon alpha (IFN-alpha), tumour necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) alpha and beta, interleukin-6 (IL-6), interleukin-8 (IL-8) and monocyte attracting chemokines. This paper reviews: (1) in vivo examinations of the cytokine profiles during influenza in mice, humans or swine; (2) in vivo data on the probable role of these cytokines; and (3) selected in vitro data on cytokine induction by the influenza virus. Examination of respiratory secretions of experimentally infected humans or animals revealed a brisk and concurrent rise in several of the cytokines mentioned. Moreover, peak cytokine levels directly correlated with virus replication and disease. In the mouse model, specific anti cytokine strategies have further confirmed the role of cytokines in body temperature changes, anorexia and lung inflammation. However, cytokines were clearly not the only factor contributing to disease, and they seemed to be essential for resolution of the infection. Though influenza virus was shown to induce cytokines in cell culture, in vitro experiments have also revealed conflicting data. Furthermore, the viral genes or products that are responsible for cytokine induction are unknown. Exactly this information would make important contributions to our understanding of the genetic basis of viral virulence. PMID- 10799784 TI - Systemic and mucosal immune responses to H1N1 influenza virus infection in pigs. AB - Influenza is a common respiratory disease in pigs, and since swine influenza viruses are zoonotic pathogens, they also pose human health risks. Pigs infected with influenza virus mount an effective immune response and are protected from subsequent challenge, whereas the currently available, inactivated-virus vaccine does not consistently confer complete protection to challenge. To develop and evaluate new vaccination strategies, it is imperative to fully understand the immune responses that are associated with protection following natural infection. Therefore, we have evaluated the phenotype and kinetics of immune responses to primary and re-challenge infection with H1N1 swine influenza virus in the pig. Through the use of isotype-specific antibody secreting cell ELISPOT assays, interferon-gamma ELISPOT assays and isotype-specific ELISAs on serum, nasal wash and bronchoalveolar lavage samples, we defined the humoral and cellular immune responses, both locally in the respiratory tract and systemically, to this viral infection. Virus-specific serum IgG, IgA, and HI titers all peaked 2-3 weeks after primary infection and did not substantially increase after re-challenge. The predominant virus-specific isotype in serum was IgG. Pigs responded with virus-specific IgG and IgA in both the upper (nasal washes) and lower (bronchoalveolar lavages) airways; IgA was the predominant isotype in both sites. Despite the fact that the pigs were completely protected from re-challenge, the antibody titers in the nasal washes increased. Results of the antibody-secreting cell ELISPOT assays demonstrated that the numbers of both IgG and IgA secreting cells in the nasal mucosa were dramatically higher than in any other tissue examined. In contrast, IFN-gamma secreting cells were predominantly localized to the spleen and tracheobronchial lymph nodes. These data will be helpful in the future development and evaluation of novel vaccines. PMID- 10799785 TI - Pandemic influenza is a zoonosis, as it requires introduction of avian-like gene segments in the human population. AB - Human influenza viruses manage to cause epidemics almost every year. The circulating viruses change their surface glycoproteins by accumulating mutations (antigenic drift) which results in variant viruses of the same subtype that are able to evade the immune pressure in the population. Every now and then, a completely new subtype of influenza A virus is introduced in the human population, which can result in an influenza pandemic. Pandemic human influenza viruses have been emerging for many centuries. Based on the genetic information of influenza viruses that have been isolated in this century, introduction of genes of the avian influenza virus reservoir obviously is required. Interspecies transmission, via another mammalian host and reassortment of avian and human influenza viruses are potential mechanisms for such an introduction. A summary of the cases in which influenza viruses containing avian-like gene segments were introduced into the human population is presented. In three cases, such infections resulted in conjunctivitis. Influenza-like illness and even pneumonia was reported in some other infections. Finally, a mortality rate of 33% was observed in the avian influenza A (H5N1) viruses that infected 18 people in Hong Kong in 1997. Although some of these viruses fulfilled some criteria of pandemic influenza viruses, they lacked the ability to rapidly spread through the human population. PMID- 10799786 TI - Interspecies transmission of influenza viruses: H5N1 virus and a Hong Kong SAR perspective. AB - This account takes stock of events and involvements, particularly on the avian side of the influenza H5N1 'bird flu' incident in Hong Kong SAR in 1997. It highlights the role of the chicken in the many live poultry markets as the source of the virus for humans. The slaughter of chicken and other poultry across the SAR seemingly averted an influenza pandemic. This perspective from Hong Kong SAR marks the coming-of-age of acceptance of the role of avian hosts as a source of pandemic human influenza viruses and offers the prospect of providing a good baseline for influenza pandemic preparedness in the future. Improved surveillance is the key. This is illustrated through the H9N2 virus which appears to have provided the 'replicating' genes for the H5N1 virus and which has since been isolated in the SAR from poultry, pigs and humans highlighting its propensity for interspecies transmission. PMID- 10799787 TI - DNA vaccination against influenza viruses: a review with emphasis on equine and swine influenza. AB - The influenza virus vaccines that are commercially-available for humans, horses and pigs in the United States are inactivated, whole-virus or subunit vaccines. While these vaccines may decrease the incidence and severity of clinical disease, they do not consistently provide complete protection from virus infection. DNA vaccines are a novel alternative to conventional vaccination strategies, and offer many of the potential benefits of live virus vaccines without their risks. In particular, because immunogens are synthesized de novo within DNA transfected cells, antigen can be presented by MHC class I and II molecules, resulting in stimulation of both humoral and cellular immune responses. Influenza virus has been used extensively as a model pathogen in DNA vaccine studies in mice, chickens, ferrets, pigs, horses and non-human primates, and clinical trials of DNA-based influenza virus vaccines are underway in humans. Our studies have focused on gene gun delivery of DNA vaccines against equine and swine influenza viruses in mice, ponies and pigs, including studies employing co-administration of interleukin-6 DNA as an approach for modulating and adjuvanting influenza virus hemagglutinin-specific immune responses. The results indicate that gene gun administration of plasmids encoding hemagglutinin genes from influenza viruses is an effective method for priming and/or inducing virus-specific immune responses, and for providing partial to complete protection from challenge infection in mice, horses and pigs. In addition, studies of interleukin-6 DNA co administration in mice clearly demonstrate the potential for this approach to enhance vaccine efficacy and protection. PMID- 10799788 TI - Vaccines protect chickens against H5 highly pathogenic avian influenza in the face of genetic changes in field viruses over multiple years. AB - Inactivated whole avian influenza (AI) virus vaccines, baculovirus-derived AI haemagglutinin vaccine and recombinant fowlpoxvirus-AI haemagglutinin vaccine were tested for the ability to protect chickens against multiple highly pathogenic (HP) H5 AI viruses. The vaccine and challenge viruses, or their haemagglutinin protein components, were obtained from field AI viruses of diverse backgrounds and included strains obtained from four continents, six host species, and isolated over a 38-year-period. The vaccines protected against clinical signs and death, and reduced the number of chickens shedding virus and the titre of the virus shed following a HP H5 AI virus challenge. Immunization with these vaccines should decrease AI virus shedding from the respiratory and digestive tracts of AI virus exposed chickens and reduce bird-to-bird transmission. Although most consistent reduction in respiratory shedding was afforded when vaccine was more similar to the challenge virus, the genetic drift of avian influenza virus did not interfere with general protection as has been reported for human influenza viruses. PMID- 10799789 TI - Equine influenza vaccine efficacy: the significance of antigenic variation. AB - To investigate the level of cross-protection induced by equine influenza H3N8 vaccines derived from different lineages, two studies have been carried out with ponies vaccinated with 'American-like' and 'European-like' vaccines and experimentally challenged with a European-like strain. The results demonstrated that equine influenza vaccines clearly protect against challenge with homologous virus if serum antibody titres are sufficiently high. On the other hand, protection is incomplete even when animals vaccinated with heterologous strains have comparative antibody levels. Nevertheless, the protection afforded by heterologous viruses can be improved by stimulating high levels of antibody. It would be advisable to update equine influenza vaccine strains regularly so that they contain similar strains to variants that are circulating in the field. PMID- 10799790 TI - A comparison of pain and its treatment in advanced dementia and cognitively intact patients with hip fracture. AB - Advanced dementia patients may be at substantial risk for undetected or undertreated pain. To examine the treatment of pain following hip fracture, a prospective cohort study was conducted in an academic teaching hospital. Fifty nine cognitively intact elderly patients with hip fracture and 38 patients with hip fracture and advanced dementia were assessed daily. The cognitively intact patients rated their pain on a numeric scale ranging from 0 (none) to 4 (very severe). Analgesics prescribed and administered were recorded and compared to hip fracture patients with advanced dementia. The advanced dementia patients received one-third the amount of morphine sulfate equivalents as the cognitively intact patients. Forty-four percent of cognitively intact individuals reported severe to very severe pain preoperatively and 42% reported similar pain postoperatively. Half the cognitively intact patients who experienced moderate to very severe pain were prescribed inadequate analgesia for their level of pain. Eighty-three percent of cognitively intact patients and 76% of dementia patients did not receive a standing order for an analgesic agent. These data reveal that a majority of elderly hip fracture patients experienced undertreated pain. The fact that advanced dementia patients received one-third the amount of opioid analgesia as compared to cognitively intact subjects-40% of whom reported severe pain postoperatively-suggests that the majority of dementia patients were in severe pain postoperatively. This study and others suggest that directed interventions to improve pain detection and alter physician prescribing practices in the cognitively impaired are needed. PMID- 10799791 TI - Symptomatology and loss of physical functioning among geriatric patients with lung cancer. AB - In this study of 129 geriatric patients with lung cancer, we investigated how symptom severity varied according to treatment type, stage of disease, and gender; how change in physical functioning (prediagnosis versus post-hospital discharge) was predicted by symptomatology, prior physical functioning, comorbidity, and age; and whether differences exist according to stage of disease, treatment status, or gender. Data were gathered through patient interviews and audits of patient records. Analysis of variance (ANOVA) techniques revealed that there were no significant differences in average symptom severity scores by gender, treatment categories, or stages of disease. Significant predictors of loss of physical functioning were symptom severity, prior physical functioning and patient age. Characteristics of a profile for elderly lung cancer patients at high risk of suffering substantial losses in physical functioning include higher prior levels of physical functioning, higher levels of current symptomatology, and lower age. PMID- 10799792 TI - Outcome of malignant spinal cord compression at a cancer center: implications for palliative care services. AB - One hundred sixty-six patients presented to a cancer center with malignant spinal cord compression (SCC) proven by magnetic resonance imaging (MRI). The majority of patients (92%) were treated with radiotherapy. Changes in functional capability over time were assessed using performance (PS) and neurological status (NS). Over the course of treatment, there was no significant change in PS or NS. The median survival from confirmation of SCC was 82 days (range 1-1349 days). Survival was significantly better for those presenting with good functional status. One hundred thirteen patients (68%) were discharged from hospital; 88 (78%) were discharged home, 11% were sent to another hospital, 4% were transferred to a rehabilitation unit, and 5% went to a hospice. Fifty-three patients (32%) died before discharge. The confirmation that PS and NS have prognostic significance in the functional outcome of patients with SCC may prove helpful in decisions regarding care planning for individual patients with SCC who are discharged from hospital. PMID- 10799793 TI - A titrated morphine analgesic regimen comparing substance users and non-users with AIDS-related pain. AB - To compare morphine dosage and effectiveness in AIDS patients with/without prior substance use and pain, a prospective, open-label case series lasting 3-18 days was conducted in both outpatients and inpatients at major pain service teaching programs. Forty-four patients, 13 with prior drug use history, who had pain associated with HIV infection or its treatment were administered sustained release morphine (SRM) every 12 hours. The dose was titrated to pain relief for a period of > or =3 consecutive days (associated with < or =2 immediate-release morphine tablets per 24 hours), or until the patient discontinued from the study or completed 18 study days. Forty-four patients were enrolled (13 with a prior drug use history). Forty were evaluable for an intent-to-treat analgesia, including 11 with a drug use history. Twenty-four (6 users) completed this study. Former users and non-users were similar in demographics, baseline pain intensities, causes of pain, discontinuation, quality of life, and acceptability of therapy. Pain intensity decreased by > or =50% in both groups (P < or = 0.0001). To identify a stable dose, the dose of SRM more than doubled in former users and rose by 31% in non-users (mean final dose 177.4 mg and 84.9 mg, respectively) (P = 0.0018). Immediate-release morphine decreased in both; former users required more (P = 0.0006). These data suggest the utility of morphine for AIDS-related pain. Patients with a prior drug use history benefited but required substantially more morphine. PMID- 10799794 TI - A pilot survey of aberrant drug-taking attitudes and behaviors in samples of cancer and AIDS patients. AB - The clinical assessment of drug-taking behaviors in medically ill patients with pain is complex and may be hindered by the lack of empirically derived information about such behaviors in particularly medically ill populations. To investigate issues surrounding the assessment of these behaviors, we piloted a questionnaire based on the observations of specialists in pain management and substance abuse. This preliminary questionnaire evaluated medication use, present and past drug abuse, patients' beliefs about the risk of addiction in the context of pain treatment, and aberrant drug-taking attitudes and behaviors. This instrument was piloted in a mixed group of cancer patients (N = 52) and a group of women with HIV/AIDS (N = 111). Reports of past drug use and abuse were more frequent than present reports in both groups. Current aberrant drug-related behaviors were seldom reported, but attitude items revealed that patients would consider engaging in aberrant behaviors, or would possibly excuse them in others, if pain or symptom management were inadequate. Aberrant behaviors and attitudes were endorsed more frequently by the women with HIV/AIDS than by the cancer patients. Patients greatly overestimated the risk of addiction in pain treatment. We discuss the significance of these findings and the need for cautious interpretation given the limitations of the methodology. This early experience suggests that both cancer and HIV/AIDS patients appear to respond in a forthcoming fashion to drug-taking behavior questions and describe attitudes and behaviors that may be highly relevant to the diagnosis and understanding management of substance use among patients with medical illness. PMID- 10799795 TI - Topical diclofenac patch relieves minor sports injury pain: results of a multicenter controlled clinical trial. AB - Sports-related soft tissue injuries, such as sprains, strains, and contusions, are a common painful condition. Current treatment includes oral nonsteroidal anti inflammatory drugs (NSAIDs), which have a high incidence of intolerable gastrointestinal side effects. Topically applied drugs have the potential to act locally in the soft tissues without systemic effects. This study assessed the efficacy and safety of topical diclofenac (NSAID) patch applied directly to the painful injury site for the treatment of acute minor sports injury pain. Adult subjects (N = 222) were recruited from two communities for a multicenter, randomized, placebo-controlled, parallel design study. All subjects had suffered a painful minor sports injury within the prior 72 hours of study entry. Either a diclofenac epolamine or placebo topical patch was applied directly to the skin overlying the painful injured site twice daily for 2 weeks. Measures of pain intensity were performed in a daily diary and at clinic visits on days 3, 7, and 14. Diclofenac patch was superior to placebo patch in relieving pain. Statistical significance was seen on clinic days 3 (P = 0.036) and 14 (P = 0. 048), as well as the daily diary pain ratings at days 3, 7, and 14 (P < or =0.044). No statistically significant differences were seen in any safety or side-effect measures with the diclofenac patch as compared to the placebo patch. Diclofenac epolamine patch is an effective and safe pain reliever for treatment of minor sports injury pain. The advantages of this novel therapy include its ease of use and lack of systemic side effects. PMID- 10799796 TI - Clinical economics: calculating the cost of acute postoperative pain medication. AB - Few data are available that address the cost of postoperative pain management, although such knowledge would enhance our understanding of caregiver choices related to direct medical costs, such as type, frequency, and route of medication. This article describes the cost of postoperative pain medications before and after an educational program provided to nurses, pharmacists, and physicians in six community hospitals. Medication costs were calculated by averaging across all brands the average wholesale price of the most common dose administered in the sample for each medication. The median cost of postoperative pain medication across all days, all surgeries, was $9.46. Calculating the cost of acute postoperative pain medication suggested that cost over stay is highly influenced by the use of a few expensive medications. The relationship of medication cost to length of stay (LOS), function, and pain intensity is discussed. PMID- 10799797 TI - Difficulties in diagnosing neuropsychiatric complications of corticosteroids in advanced cancer patients: two case reports. AB - Because of their variety of uses, corticosteroids are frequently prescribed in advanced cancer patients. Two patients who developed neuropsychiatric complications on corticosteroids and their subsequent management are described. The first patient, who had a known history of steroid-induced psychotic depression, required corticosteroids to treat recurrent brain edema from a malignant meningioma. The patient was managed by using low-dose corticosteroids and concomitant haloperidol. The second patient was prescribed corticosteroids for a constellation of symptoms, including pain and nausea from a possible bowel obstruction, and developed a severe delirium that required discontinuation of the corticosteroids. The difficulties of diagnosing steroid-related cognitive and mood changes in advanced cancer patients who often have multisystem disease are discussed, as well as strategies for minimizing the effects of corticosteroids' neuropsychological complications. PMID- 10799798 TI - Characterization of the interaction of Escherichia coli heat-stable enterotoxin (STa) with its putative receptor on the intestinal tract of newborn calves. AB - Enterotoxigenic Escherichia coli (ETEC) induces severe diarrhea in newborn calves through the elaboration of heat-stable enterotoxin (STa). We investigated the distribution and characteristics of the STa-specific receptors on enterocytes and brush border membrane vesicles (BBMVs) prepared from anterior jejunum, posterior jejunum, ileum and colon of newborn calves. We found that density of the receptors and their affinity to STa were higher on enterocytes and BBMVs that were derived from the ileum than enterocytes and BBMVs prepared from other segments of the calf intestine. This study suggests that, in newborn calves, the ileum is the major part of the intestinal tract that is affected in the course of secretory diarrhea caused by STa-producing ETEC strains. PMID- 10799799 TI - Stimulation of bacterial adherence by neutrophil defensins varies among bacterial species but not among host cell types. AB - Adherence of Haemophilus influenzae to bronchial epithelial cells is enhanced by neutrophil defensins, which are released from activated neutrophils during inflammation [Gorter et al. (1998) J. Infect. Dis. 178, 1067-1078]. In this study, we showed that the adherence of H. influenzae to various epithelial, fibroblast-like and endothelial cell types was significantly enhanced by defensins (20 microg ml(-1)). Defensins stimulated also the adherence of Moraxella catarrhalis, Neisseria meningitidis and nonencapsulated Streptococcus pneumoniae to the NCI-H292 cell line. In contrast, defensins did not affect the adherence of Pseudomonas aeruginosa, encapsulated S. pneumoniae, Escherichia coli and Staphylococcus epidermidis. These results suggest that the defensin-enhanced adherence might support the adherence and possibly persistence of the selected bacterial species using the respiratory tract as port of entry. PMID- 10799800 TI - Genotyping of cagA and vacA, Lewis antigen status, and analysis of the poly-(C) tract in the alpha(1,3)-fucosyltransferase gene of Irish Helicobacter pylori isolates. AB - Much work has focused on trying to identify markers in Helicobacter pylori that might allow the eventual disease outcome of an infection to be predicted. In this study we examined the cagA and vacA genotype, and Lewis status in a panel of 43 Irish H. pylori clinical isolates, and investigated a possible correlation with disease pathology. In addition, differences in the poly-(C) tract of the alpha(1,3)-fucosyltransferase gene were examined to identify a possible correlation with gene expression. Only three of 43 isolates were cagA-negative, whereas the remaining 40 isolates, independent of pathology, were cagA-positive. In all the strains we examined, the vacA signal-sequence was type s1a. For the vacA mid-region 12/43 isolates were type m1 and 31/43 isolates were type m2. These data, and examination of isolates from different pathology groups, suggests that there is no correlation between virulence and vacA genotype in the Irish population of H. pylori isolates. Western blotting of whole cell lysates from 32 H. pylori isolates showed 3/32 displayed only the Le(x) epitope, 12/32 only the Le(y), 13/32 both epitopes and 4/32 neither epitope. No apparent association between Lewis phenotype and disease pathology was evident. A range of lengths of poly-(C) tract were observed in the alpha(1, 3)-fucosyltransferase gene, however the length of the tract in an isolate did not correlate with the Lewis structures present. We conclude that future studies on H. pylori pathogenesis should not alone focus on the importance of molecular markers, but also on the host response, including genetic background and immune responsiveness. PMID- 10799801 TI - Molecular cloning, purification and immunological responses of recombinants GroEL and DnaK from Streptococcus pyogenes. AB - To better understand the roles of heat shock proteins in streptococcal diseases, the groEL and dnaK genes from Streptococcus pyogenes were cloned and their products (GroEL and DnaK) and derivatives (F2GroEL, F3GroEL and C1DnaK) purified as His-tagged fusion proteins. Western blot analysis of the purified proteins with sera from individuals with streptococcal diseases demonstrated that 29 out of 36 sera tested were reactive with GroEL and eight recognized DnaK. Rabbit antiserum against myosin recognized both GroEL and DnaK. Antibodies raised against purified F2GroEL and DnaK reacted with myosin in the ELISA but not in a Western immunoblot. These data indicate that the S. pyogenes GroEL and DnaK may be important immunogens during streptococcal infections. Furthermore, we provide evidence of an immunogenic relatedness of the GroEL and DnaK proteins with myosin that could play a role in the pathogenesis of streptococcal non-suppurative sequelae. PMID- 10799802 TI - Interaction of clinical isolates of nonencapsulated Haemophilus influenzae with mammalian extracellular matrix proteins. AB - The adherence of clinical isolates of nonencapsulated Haemophilus influenzae strains from patients with chronic bronchitis to distinct immobilized extracellular matrix components was determined. With selected strains the induction of plasmin formation by these isolates was studied. The strains could be divided into two groups: strains that showed a very high level of adherence to laminin and type I collagen, as well as adhesion to fibronectin and strains that showed only a moderate level of adhesion to laminin and a low level of adhesion to fibronectin. Plasmin formation was demonstrated for three out of eight isolates. Persisting and nonpersisting strains did not differ quantitatively or qualitatively with respect to the level of adhesiveness to the distinct matrix proteins and in their ability to induce plasmin formation. PMID- 10799803 TI - Determination of the immunodominant part in the O-antigenic polysaccharide from Escherichia coli O128 by ELISA-inhibition study. AB - The immunodominant part in the O-antigenic polysaccharide from Escherichia coli O128 was immunologically characterized by an enzyme-linked immunosorbent assay (ELISA). The antibody specificity was determined by the inhibitory effects of the methyl glycosides of constituent mono- and oligosaccharides synthesized related to the O-antigenic polysaccharide from E. coli O128. It was found that methyl alpha-L-fucopyranoside was the most effective inhibitor amongst the monosaccharides while the highest antibody specificity was directed towards the trisaccharide with the structure: beta-D-GalpNAc-(1-->6)-[alpha-L-Fucp-(1-->2)] beta-D-Galp-1-->OMe suggesting that the monospecific antibody has the extended combining site. PMID- 10799804 TI - IgG antibody titer against Helicobacter pylori correlates with presence of cytotoxin associated gene A-positive H. pylori strains. AB - The level of the IgG antibody titer against Helicobacter pylori correlates with the severity of gastritis. H. pylori strains can harbor the so-called pathogenicity island, containing the cytotoxin associated gene (cagA). Since cagA positive strains are more virulent it can be postulated that the gastritis will be more severe and hence the IgG antibody titer higher. In a cross-sectional study the correlation of IgG antibody titer and cagA status was studied from patients undergoing upper gastrointestinal endoscopy. Biopsy specimens were obtained to determine the H. pylori status. In addition a serum sample was taken for detection of IgG antibodies against H. pylori as well as CagA. A total of 290 patients positive for IgG antibodies against H. pylori were included. Of these 153 were cagA-positive and 137 were cagA-negative. The mean IgG antibody titer was significantly higher in cagA-positive patients compared to cagA-negatives, 0.75 (S.D. 0.22) versus 0.69 (S.D. 0.24) (P=0.033). It is concluded that the IgG antibody titer is significantly higher in patients harboring cagA-positive H. pylori strains. However, in daily practice the level in IgG antibody titer cannot predict whether or not an individual carries a cagA-positive H. pylori strain since major overlap in IgG antibody titer between cagA-positive and cagA-negative patients is present. PMID- 10799805 TI - Clinical relevance and virulence factors of pigmented Serratia marcescens. AB - Pigmented Serratia marcescens isolated in a Brazilian hospital were studied with respect to frequency of isolation, serotyping, antibiotic resistance and virulence factors. The serotype most frequent was O6:K14 (53%) and all isolates were resistant to ampicillin, cephalothin and tetracycline. The majority of the isolates (92%) were resistant to the action of human serum and all produced cytotoxins on Vero, CHO, HEp-2 and HeLa cells. These isolates were virulent for mice (LD(50)=10(7) bacteria ml(-1)) and showed virulence factors, but were isolated with low frequency (3. 4%) and caused infection in only 31% of cases. Analysis of serotyping, phage typing and chromosomal DNA revealed at least 13 unrelated strains among pigmented S. marcescens. In conclusion, this work describes a low frequency of isolation of pigmented S. marcescens from clinical specimens, indicating that non-pigmented strains are clinically more significant. PMID- 10799806 TI - Role of interleukin-12 in patients with dengue hemorrhagic fever. AB - Interleukin (IL)-12 has a broad range of activities including regulation of cytokine synthesis and selective promotion of Th1-type cell development. A shift from a Th1-type response to Th2-type has been suggested to be important in the pathogenesis of dengue hemorrhagic fever (DHF). This study was undertaken to investigate the possible role of IL-12 in this shift. A total of 76 patients with various grades of dengue illness and 21 normal healthy controls were tested for IL-12 levels in serum samples and IL-12 mRNA in their peripheral blood mononuclear cells. The results showed that the levels of IL-12 were the highest in patients with dengue fever (270+/-102 pg ml(-1)) followed by decreasing levels in the patients with DHF grade I (198+/-86 pg ml(-1); P<0.05) and DHF grade II (84+/-52 pg ml(-1); P<0.001). Neither IL-12 nor its mRNA could be detected in the patients with DHF grades III and IV. The cytokine appeared and reached peak levels during the first 4 days of illness, started to decline by day 5-8 and disappeared by day 9 onwards. The absence of IL-12 during severe illness and late phases of the disease may be responsible for the shift to a Th2-type response and thus for the pathogenesis of DHF. PMID- 10799807 TI - Enhancement of IL-8 production from human monocytic and granulocytic cell lines, THP-1 and HL-60, stimulated with Malassezia furfur. AB - Previously, we reported that Malassezia furfur, causing systemic fungal infection, was taken up into human monocytic cell line, THP-1, in a concentration dependent manner. This fact suggested that M. furfur could activate phagocytes, such as monocyte and polymorphonuclear leukocyte. Thus we examined cytokine mRNA expression from human monocytic and granulocytic cell line, THP-1 and HL-60, stimulated with M. furfur by using reverse transcriptase-polymerase chain reaction (RT-PCR) and ELISA. We chose IL-1alpha, IL-6, IL-8, IL-12 and TNF-alpha as primers for THP-1, and IL-1alpha, IL-6 and IL-8 for HL-60. M. furfur induced the expression of IL-8 mRNA from THP-1 and HL-60 following incubation for 3 h, and also induced IL-1alpha mRNA from HL-60, although this induction was weaker than that of IL-8 mRNA. Furthermore, opsonized M. furfur induced stronger expression of IL-8 mRNA in comparison with intact M. furfur. IL-8 production from THP-1 and HL-60 was enhanced in a concentration- and incubation time-dependent manner. These facts strongly suggested that M. furfur could activate phagocytes, and could induce inflammatory responses in systemic infection. PMID- 10799808 TI - Structure and serologic properties of O-specific polysaccharide from Citrobacter freundii possessing cross-reactivity with Escherichia coli O157:H7. AB - Citrobacter freundii OCU158 is a serologically cross-reactive strain with Escherichia coli O157:H7. To explore the close relationship between two strains, we have analyzed the chemical structures of O-specific polysaccharides and antigenic properties of lipopolysaccharides (LPSs) of both strains. The structure of O-specific polysaccharides from both strains was found to be identical by chemical and nuclear magnetic resonance analyses, in which D-PerNAc was 4 acetamido-4,6-dideoxy-D-mannose: [-->4)-beta-D-Glc-(1-->3)-alpha-D-PerNAc-(1-->4) alpha-D-GalNAc-(1 --> 3)-alpha-L-Fuc-(1-->](n). The enzyme immunoassay using LPS derived either from E. coli O157 or from C. freundii could equally detect high levels of serum antibodies against LPS in patients with enterohemorrhagic E. coli (EHEC) O157 infection. Absorption of antibodies in EHEC patient serum by LPS from E. coli O157 or C. freundii, however, showed a difference in the epitopes. This difference was attributable to the epitope specificity of the core region and/or lipid A structure in LPS. PMID- 10799809 TI - Evaluation of the Applied Biosystems automated Taqman polymerase chain reaction system for the detection of meningococcal DNA. AB - In a period where the proportion of culture confirmed cases in the UK has been steadily declining, diagnosis by PCR has been used to increase the number of confirmed cases and provide additional epidemiological data. This report presents a comparative evaluation of the fluorogenic probe-based 5' exonuclease assay (Taqman) using the Perkin-Elmer Applied Biosystems automated sequence detection system 7700 with previously reported polymerase chain reaction enzyme-linked immunosorbent (PCR ELISA) assays for the detection of meningococcal DNA in CSF, plasma and serum samples. Taqman assays developed were based on the detection of a meningococcal capsular transfer gene (ctrA), the insertion sequence IS1106 and the sialytransferase gene (siaD) for serogroup B and C determination and compared with similar assays in a PCR ELISA format. The Taqman ctrA assay was specific for Neisseria meningitidis, however the IS1106 assay gave false positive reactions with a number of non-meningococcal isolates. Sensitivity of the Taqman ctrA, IS1106 and siaD assays testing samples from culture-confirmed cases were 64, 69 and 50%, respectively, compared with 26, 67 and 43% for the corresponding PCR ELISA assays. Improvements to the DNA extraction procedure has increased the sensitivity to 93 and 91% for the TaqMan ctrA and siaD assays, respectively, compared to culture confirmed cases. Since the introduction of Taqman PCR a 56% increase in laboratory confirmed cases of meningococcal disease has been observed compared to culture only confirmed cases. The developed Taqman assays for the diagnosis of meningococcal disease enables a high throughput, rapid turnaround of samples with considerable reduced risk of contamination. PMID- 10799810 TI - Enhanced antisense efficacy of oligonucleotides adsorbed to monomethylaminoethylmethacrylate methylmethacrylate copolymer nanoparticles. AB - The purpose of this study was the investigation of cationic nanoparticles as drug delivery systems for antisense oligonucleotides. Cationic monomethylaminoethylmethacrylate (MMAEMA) copolymer nanoparticles were prepared from N-monomethylaminoethylmethacrylate hydrochloride and methylmethacrylate. Oligonucleotides were adsorbed onto MMAEMA nanoparticles. Cell penetration was investigated in vitro with fluorescently labeled oligonucleotides and nanoparticles. Antisense effects of oligonucleotides adsorbed to MMAEMA nanoparticles were evaluated by sequence specific inhibition of ecto-5' nucleotidase expression. The amount of enzyme expressed in PC12 cells was detected and quantified by immunocytochemistry using fluorescein isothiocyanate labeled antibodies. Oligonucleotides were adsorbed to MMAEMA nanoparticles by the formation of ion-pairs between the positively charged secondary amino groups located on the particle surface and the anionic phosphodiester or phosphorothioate backbones of the oligonucleotides. Adsorption to nanoparticles led to an increased cellular uptake of oligonucleotides and to a significantly enhanced antisense efficacy of unmodified phosphodiester oligonucleotides as well as phosphorothioates. The results of the cell penetration and the antisense assay demonstrated that MMAEMA nanoparticles are promising carriers for oligonucleotide administration. PMID- 10799811 TI - Vitamin A loaded solid lipid nanoparticles for topical use: occlusive properties and drug targeting to the upper skin. AB - To evaluate the potential use of solid lipid nanoparticles (SLN) in dermatology and cosmetics, glyceryl behenate SLN loaded with vitamin A (retinol and retinyl palmitate) and incorporated in a hydrogel and o/w-cream were tested with respect to their influence on drug penetration into porcine skin. Conventional formulations served for comparison. Excised full thickness skin was mounted in Franz diffusion cells and the formulations were applied for 6 and 24 h, respectively. Vitamin A concentrations in the skin tissue suggested a certain drug localizing effect. High retinol concentrations were found in the upper skin layers following SLN preparations, whereas the deeper regions showed only very low vitamin A levels. Because of a polymorphic transition of the lipid carrier with subsequent drug expulsion following the application to the skin, the drug localizing action appears to be limited for 6-24 h. Best results were obtained with retinol SLN incorporated in the oil-in-water (o/w) cream retarding drug expulsion. The penetration of the occlusion sensitive drug retinyl palmitate was even more influenced by SLN incorporation. Transepidermal water loss (TEWL) and the influence of drug free SLN on retinyl palmitate uptake exclude pronounced occlusive effects. Therefore enhanced retinyl palmitate uptake should derive from specific SLN effects and is not due to non-specific occlusive properties. PMID- 10799812 TI - Characterization of the physicochemical properties of the micelles by the novel platelet activating factor receptor antagonist E5880. AB - E5880, a novel platelet activating factor receptor antagonist, was dispersed in the buffer solution (4.8 mM citric acid, 10% lactose, pH 2.8) for the preparation of an injectable formulation and the physicochemical properties of the micelles were characterized. The critical micelle concentration of E5880 was 0.12 mM. Using the area per molecule results, the critical packing parameter was calculated and showed that the structure was spherical and the number of molecules in the aggregates was 46. The diameter of the micelle was 5.6 nm. The micropolarity around the hydrocarbon region of the micelle was similar to that of isobutanol. PMID- 10799813 TI - A five way crossover human volunteer study to compare the pharmacokinetics of paracetamol following oral administration of two commercially available paracetamol tablets and three development tablets containing paracetamol in combination with sodium bicarbonate or calcium carbonate. AB - This report concerns a single dose randomized five way crossover study to compare the pharmacokinetics of paracetamol from two commercially available paracetamol (500 mg) tablets and three different development paracetamol (500 mg) tablet formulations containing either sodium bicarbonate (400 mg), sodium bicarbonate (630 mg) or calcium carbonate (375 mg). The results demonstrated that addition of sodium bicarbonate (630 mg) to paracetamol tablets, increased the rate of absorption of paracetamol relative to conventional paracetamol tablets and soluble paracetamol tablets. Addition of sodium bicarbonate (400 mg) to paracetamol tablets increased the absorption rate of paracetamol relative to conventional paracetamol tablets, but there was no difference in the rate of absorption compared to soluble paracetamol tablets. Inclusion of calcium carbonate (375 mg) to paracetamol tablets had no effect on absorption kinetics compared to the conventional paracetamol tablet. The faster absorption observed for the sodium bicarbonate formulations may be as a result of an increase in gastric emptying rate leading to faster transport of paracetamol to the small intestine where absorption takes place. PMID- 10799814 TI - Hydroxypropylmethylcellulose (HPMC) as emulsifier for submicron emulsions: influence of molecular weight and substitution type on the droplet size after high-pressure homogenization. AB - Hydroxypropylmethylcellulose (HPMC) is a known emulsifier as well as a common viscosity enhancer in eye drops. Therefore, HPMC stabilized emulsions appear as interesting drug carriers for ophthalmic use and as a suitable treatment of dry eye syndrome. Since submicron emulsions are known to have an improved drug delivery, attempts were made to reduce the emulsion's droplet size by high pressure homogenization. Droplet size was dependent on the homogenization pressure and the polymer content. Differences were found between emulsions stabilized with higher and lower molecular weight HPMC. Smaller droplet sizes were obtained with the shorter chained HPMC. No considerable influence of the substitution type was observed. PMID- 10799815 TI - Melt extrusion--an alternative method for enhancing the dissolution rate of 17beta-estradiol hemihydrate. AB - 17Beta-estradiol hemihydrate (17beta-E2) is a poorly water-soluble drug. Physical methods for improving the solubility and dissolution rate, e.g. micronization, have certain inherent disadvantages. The method of choice in this study, melt extrusion, proved to overcome many of the shortcomings of conventional methods. Different compositions of excipients such as PEG 6000, PVP (Kollidon 30) or a vinylpyrrolidone-vinylacetate-copolymer (Kollidon VA64) were used as polymers and Sucroester WE15 or Gelucire 44/14 as additives during melt extrusion. The solid dispersions resulted in a significant increase in dissolution rate when compared to the pure drug or to the physical mixtures. For example, a 30-fold increase in dissolution rate was obtained for a formulation containing 10% 17beta-E2, 50% PVP and 40% Gelucire 44/14. The solid dispersions were then processed into tablets. The improvement in the dissolution behavior was also maintained with the tablets. The USP XXIII requirement for estradiol tablets reaching greater than 75% drug dissolved after 60 min was obtained in this investigation. PMID- 10799816 TI - Influence of processing and curing conditions on beads coated with an aqueous dispersion of cellulose acetate phthalate. AB - The influence of fluidized-bed processing conditions, as well as curing parameters with and without humidity, on drug release from beads coated with cellulose acetate phthalate (CAP) aqueous dispersion was investigated. Theophylline beads prepared by extrusion-spheronization were coated with diethyl phthalate (DEP)-plasticized CAP dispersion (Aquacoat CPD) using a Strea-1 fluidized-bed coater. The parameters investigated were plasticizer level, outlet temperature, spray rate during coating application and fluidizing air velocities using a half-factorial design. The processing temperature during coating applications was identified as a critical factor among the variables investigated. The release rate significantly decreased when the beads were coated at 36 degrees C compared to those coated at 48 degrees C (P<0.01). Higher coating efficiencies and better coalescence of films were obtained at the lower coating temperature. Above the minimum film-formation temperature (MFFT), drug release in acid decreased as the coating temperature was decreased. Curing at 60 degrees C significantly reduced the drug release for beads coated at 32 degrees C, but had no significant effect on drug release for beads coated at temperatures above 36 degrees C. Curing at 50 degrees C in an atmosphere containing 75% RH (relative humidity), irreversibly converted poor film formation into better coalescence, and increased the mechanical toughness of films. Subsequent removal of the moisture absorbed from beads did not significantly alter the enteric profiles obtained through heat-humidity curing. The extent of coalescence via heat humidity curing was dependent on the curing temperature, % humidity, curing time and coating temperature. The results demonstrated the importance of the selection of coating temperature for CAP-coated beads and the role of moisture on CAP film formation. Curing with humidity was found to be more effective than without. PMID- 10799818 TI - Physicochemical properties of amorphous precipitates of cimetidine-indomethacin binary system. AB - We have found that the binary system, consisting of a precipitate of cimetidine and naproxen, became amorphous due to intermolecular interaction. In order to clarify the interaction between cimetidine and other drugs, the physicochemical properties of binary systems consisting of cimetidine and drugs, phenacetin, salicylamide or indomethacin, were investigated. X-ray powder diffraction patterns and thermal analysis findings for the precipitates indicated that the cimetidine-indomethacin system has an amorphous structure, whereas the cimetidine phenacetin and cimetidine-salicylamide systems do not. Fourier-transform infra red (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy findings suggested that there is an intermolecular interaction between a proton in the imidazole ring of cimetidine and the C=O in the COOH of indomethacin. Since an interaction by the hydrogen bond between cimetidine and indomethacin would prevent three-dimensional arrangements of the molecules, the precipitate would be amorphous. In the cimetidine-indomethacin system, decarboxylation of indomethacin occurred below the melting temperature, indicating that the chemical stability decreased upon precipitation. Cimetidine was found to interact with drugs with a carboxyl group. The interaction would be applicable to make the amorphous system of the drugs and increase the solubility of the drugs. PMID- 10799817 TI - Formation of peptide impurities in polyester matrices during implant manufacturing. AB - Most peptides are susceptible, in vivo, to proteolytic degradation, and it is difficult to formulate and to deliver them without loss of biological activity. In addition, it is often desirable to release them continuously and at a controlled rate over a period of weeks or months. For these reasons, a controlled release system is suitable. Poly(lactic acid) (PLA) is a biocompatible and biodegradable material that can be used for many applications, including the design of injectable controlled release systems for pharmaceutical agents. Development of these delivery systems presents challenges in the assessment of stability, specially for peptide drugs. By means of an extrusion method, long acting poly(lactic acid) implants containing vapreotide, a somatostatin analogue, were prepared. The nature of the main degradation product obtained after implant manufacturing was elucidated. It was found that the main peptide impurity was a lactoyl lactyl-vapreotide conjugate. Because lactide are found in small quantities in most commercially available PLA, the influence of residual lactide in the polymeric matrix, on the formation of peptide impurities during manufacturing, was specially investigated. This work demonstrates that the degree of purity of the carrier is of great importance with regard to the formation of peptide impurities. PMID- 10799819 TI - A new theoretical model to characterize the densification behavior of tableting materials. AB - The purpose of the study was to develop a new three-dimensional model using force, time and displacement to characterize the densification behavior of tableting materials. Normalized time (x), displacement converted to ln(1/1 - D(rel)) according to Heckel (y) and force presented as pressure (z) were used to plot a graph. A twisted plane was fitted to this three-dimensional plot. This plane was characterized by three parameters d, the slope over time called 'time plasticity', e, the slope over pressure called 'pressure plasticity' and omega, the angle of rotation called 'fast elastic decompression'. These parameters were used to characterize the densification behavior of the well-known materials microcrystalline cellulose, dicalcium phosphate dihydrate, theophylline monohydrate, cellulose acetate and hydroxypropyl methylcellulose at different rho(rel, max). It could be shown that brittle, elastic and plastic compression properties could be very well distinguished and differentiated. Further on, it could be shown whether these properties were due to pressure or time. Thus this model has the prevailing advantage to characterize tableting materials in one step according to time and pressure and it is a useful tool to develop tablet formulations or new excipients. PMID- 10799820 TI - Development of computerised procedures for the characterisation of the tableting properties with eccentric machines: extended Heckel analysis. AB - Heckel plots are a suitable and valuable method for analysis of powder compaction with very small amounts of powder. The determination is based upon a non-linear transformation of compression data and thus the signal errors that might be introduced into the analysis might be enlarged and become critical. The method of determination of true density affects the results dramatically as does the accuracy of the powder height determination. The porosity should be corrected for compression of the solid fraction. The accuracy of the powder height detection is the most demanding parameter. The statements are proven by simulations based on real data and analytic calculation. According to these highly corrected Heckel plots, the shape of the plots during the compression phase gives the information about fragmentation and plasticity and additionally about the time dependency of the compression behaviour within one compression on an eccentric press. PMID- 10799821 TI - Modeling of the drug delivery from a hydrophilic transdermal therapeutic system across polymer membrane. AB - A mathematical simulation is presented which describes the in vitro drug delivery kinetics from hydrophilic adhesive water-soluble poly-N-vinylpyrrolidone (PVP) polyethylene glycol (PEG) matrices of transdermal therapeutic systems (TTS) across skin-imitating hydrophobic Carbosil membranes. Propranolol is employed as the test drug. The contributions of the following physicochemical determinants to drug delivery rate control have been estimated: the drug diffusion coefficients both in the matrix and the membrane; the membrane-matrix drug partition coefficient: the drug concentration in the matrix and the membrane thickness. Drug transfer from the hydrophilic matrix across the membrane is shown to be controlled by the drug partitioning from the matrix into the membrane. The best correlation between simulation data and experimental results is obtained when the effect of membrane hydration is taken into consideration during in vitro drug release. PMID- 10799822 TI - Equivalence testing and equivalence limits of metered-dose inhalers and dry powder inhalers measured by in vitro impaction. AB - In this study, criteria for the acceptability of comparative in vitro equivalence testing are proposed. Furthermore, the following equivalence limits for in vitro impaction methods are postulated: the 90% confidence interval (CI) of the in vitro deposition ratio of the test product and the reference product should lie within 0.80-1.20. The aim of this study was to challenge these limits by applying them to in vitro impaction results of several groups of pressurized metered-dose inhalers and dry powder inhalers containing salbutamol and beclomethasone dipropionate. The deposition results were obtained with the Twin Impinger. All products had a marketing authorization in The Netherlands and were considered therapeutically equivalent within each group. The postulated equivalence limits/group were challenged by fictitiously assigning a preparation as a test product or reference product and calculating the 90% CI of the deposition ratio of the test and reference products. All possible combinations of products within a group were tested. The products were considered equivalent if the 90% CI of the quotient lay within 0.80-1.20. In most cases, the quotient of the test product and reference product remains within 0.80-1.20, but due to a high variability in the deposition results of several products, the 90% CI of the quotient sometimes falls outside the proposed equivalence limits. It is concluded that the equivalence limits postulated are rather conservative, with respect to accepting equivalence. The limits can therefore serve as a prudent predictor of equivalence within the acceptability criteria proposed, but have to be further validated. PMID- 10799823 TI - Preparation of avidin-labeled protein nanoparticles as carriers for biotinylated peptide nucleic acid. AB - The possibility of preparing protein nanoparticles followed by covalent linkage of avidin was investigated. Free sulfhydryl groups were introduced onto the surface of protein nanoparticles either by aldehyde quenching with cysteine or reaction of free amino groups with 2-iminothiolane. The number of primary amino groups and sulfhydryl groups on the surface of the resulting particles was quantified with site-specific reagents. Avidin was attached to the surface of the thiolated nanoparticles via a bifunctional spacer which reacted in a first step with amino groups of avidin and in a second step with the sulfhydryl groups introduced onto the surface of the nanoparticles. Biotinylated peptide nucleic acid (PNA) as a model compound for biotinylated drugs was effectively coupled to the nanoparticles by complex formation with the covalently attached avidin. Since the formation of the interaction between biotin and avidin is very rapid and stable a highly effective drug carrier system for biotinylated compounds such as PNAs was achieved. PMID- 10799824 TI - Ruptured thoracoabdominal aortic aneurysm repaired with minimal aortic occlusion time and continuous visceral perfusion - a new technique. PMID- 10799825 TI - Emergency surgery of acute traumatic arteriovenous fistulas. AB - The purpose of this report was to analyze the clinical presentation, diagnosis, surgical and non-surgical treatment and the outcome of patients with acute AVFs seen during the last 8years. During 1991-98 we treated 47 patients with traumatic AVFs. Among these patients 11 were classified as 'acute' AVF. All the patients were male and they ranged in age from 17 to 64yr (mean 25yr). The time from injury to admission to our two institutions varied from 4h to 17days. Emergency surgery was performed in unstable patients and in those with an expanding haematoma base on clinical assessment alone. The acute surgery group consisted of stable patients that were operated after angiography examination. A ??? murmur and thrill were present in seven patients, and peripheral pulses were absent in four patients. The main blood vessels were reconstructed using various techniques in eight patients and minor blood vessel were ligated in three patients. Two lower limb amputations had to be performed, both after reconstruction of popliteal AVFs. In the remaining nine patients reconstruction of the axial vessels was successful and no signs of ischemia developed in patients after ligation of minor vessels. No neurological deficit developed in two patients whose AVFs were vaporised in the neck. The average hospital stay was 8. 6days. Emergency surgery is safe in unstable patients with traumatic AVFs. A thrill and murmur are characteristic signs even in the acute setting, but the examiner must be persistent in looking for them whenever there is a suspicion of a blood vessel injury. Angiography is a reliable diagnostic tool in stable patients, but whether it is essential when there is an expanding pulsating hematoma remains debatable. PMID- 10799826 TI - Early assessment of skeletal muscle damage after ischaemia-reperfusion injury using Tc-99m-glucarate. AB - PURPOSE: After acute arterial obstruction of the lower extremity, muscle damage is the critical determinant for clinical outcome. The extent of muscle damage and limb viability are currently assessed by clinical examination, which is inaccurate. Tc-99m-pyrophosphate (PYP) has been applied for imaging ischaemia reperfusion damage. More recently, a new imaging agent, Tc-99m-glucarate (GLUC), was introduced for delineating early myocardial infarction after ischaemia reperfusion. The aim of this study was to determine if GLUC could delineate early skeletal muscle damage after ischaemia-reperfusion. Both tracers were used in a novel murine model of hindlimb ischaemia-reperfusion. METHODS: In anaesthetised mice, ischaemia of one hindlimb was maintained for 2, 3 and 4h using a tourniquet, followed by a reperfusion period of 1h. Additionally, reperfusion periods of 3, 24 and 96h were studied after 3h of ischaemia. PYP or GLUC was injected 45min before end of reperfusion. Concentrations of both agents were determined in blood, reperfused and contralateral muscle. Reperfused-to contralateral muscle ratios were calculated. In separate experiments, muscle biopsies were obtained for histologic examination. RESULTS: Ischaemia and reperfusion damage was demonstrated histologically. Using scintigraphy GLUC depicted reperfusion significantly better than PYP. After 2, 3 and 4h of ischaemia, the reperfused-to-contralateral ratios for GLUC were 10.7+/-0.9, 8.9+/ 0.9 and 8.6+/-1.1, as compared to 4.5+/-0.7, 4.9+/-0.4 and 4.5+/-0.4 for PYP (P<0.05 at all points). For longer periods of reperfusion, the ratios for GLUC decreased to similar levels as observed for PYP. CONCLUSION: The present study indicates that GLUC is a specific early marker of myocyte necrosis after ischaemia-reperfusion. GLUC may become an useful agent for clinical, early, non invasive monitoring of skeletal muscle damage after ischaemia-reperfusion. PMID- 10799828 TI - A review of myocardial normothermia: its theoretical basis and the potential clinical benefits in cardiac surgery. AB - Myocardial protection during cardiac surgery aims to preserve myocardial function while providing a bloodless and motionless operating field to make surgery easier. Myocardial protection is achieved by decreasing the oxygen needs using hypothermia and producing electromechanical cardiac arrest using potassium infusion which allows surgery to be performed on a non-beating heart. The deleterious effects of hypothermia include dysfunction of enzymatic systems, development of acidosis, a decrease in tissue oxygen delivery, an increase in blood viscosity and a decrease in erythrocyte deformability. Ninety percent of the decrease in oxygen consumption is obtained by inducing electromechanical arrest and inducing hypothermia has little additional benefit. Maintenance of systemic and myocardial normothermia reduces problems and provides a more physiological approach for cardiopulmonary bypass (CPB). The current results obtained using normothermic protection are very encouraging, and it is an easier inexpensive option. This review summarizes the current knowledge on the benefits of normothermia, based upon experimental and clinical studies. PMID- 10799827 TI - The effect of captopril on membrane bound enzymes in ischemia-reperfusion injury. AB - There is substantial evidence that Na(+)K(+)/Mg(2+) ATPase and Ca(2+)/Mg(2+) ATPase enzymes would effect the membrane integrity. Forty guinea pig (n=10 in each group) hearts were studied in an isolated Krebs-Henseleit solution perfused Langendorff cardiac model. The first group was utilized as the control group. Group 2 hearts were arrested with captopril (200micromol/l) added St Thomas Hospital Cardioplegic Solution (STHCS). Group 3 animals were pretreated with oral captopril (0.3mg/kg/twice a day) for 10days and then arrested with STHCS. Group 4 hearts were again pretreated with oral captopril (0.3mg/kg/twice a day for 10days) arrested with STHCS and reperfused with captopril added Krebs-Henseleit solution (200micromol/l). Hearts were subjected to normothermic global ischemia for 90min and than were reperfused at 37 degrees C. When the treated groups were compared with control, best results were achived by group 4. The Na(+)K(+) and Ca(2+)/Mg(2+) ATPase levels increased from 466.38+/-5.99 to 564.13+/-7.77 and 884.69+/-9.13 to 1254.29+/-5.75 nmol Pi/mg/prot/h respectively (P<0.05). These results suggest that captopril protects the membrane integrity and thus played a role at the recovery of depressed membrane bound Na(+)K(+)/Mg(2+) ATPase and Ca(2+)/Mg(2+) ATPase activity and also in ischemia-reperfusion injury. PMID- 10799829 TI - Haemodynamic changes during minimally invasive coronary artery bypass surgery using high-dose esmolol. AB - We aimed to investigate the effects of high-dose esmolol on haemodynamics and oxygen extraction in minimally invasive direct coronary artery bypass (MIDCAB) surgery patients. METHODS: In 18 patients, heart rate (HR), mean arterial (MAP), central venous (CVP), pulmonary capillary wedge pressure (PCWP), cardiac output (CO), and mixed venous oxygen saturation (Sv0(2)) were prospectively measured after induction of anaesthesia (T1), start of surgery (T2), during bypass grafting with beta-blockade (T3), and at the end of surgery (T4). RESULTS: Mean esmolol dose at T3 was 0.44+/-0.2mgkg(-1)min(-1). HR was unchanged, whereas significant decreases in mean CO (3.1+/-0. 8 vs 4.8+/-1.0lmin(-1)m(-2), pre esmolol), MAP (53+/-10 vs 89+/-14mmHg), and SvO(2) (65+/-10 vs 81+/-4%) were observed during esmolol administration. All haemodynamic parameters normalized immediately after termination of esmolol (T4). CONCLUSIONS: Despite unchanged HR esmolol reduced CO and MAP suggesting a favorable reduction of myocardial oxygen consumption. Mean Sv0(2) during esmolol administration reflects an acceptable ratio of whole-body oxygen delivery and consumption. Haemodynamic changes with high-dose esmolol during MIDCAB surgery remain within safety margins. PMID- 10799830 TI - Active infective endocarditis: low mortality associated with early surgical treatment. AB - BACKGROUND: Early surgical treatment is important for successful outcome in selected cases of active, either native (NVE) or prosthetic valve endocarditis (PVE). The aim of this study was to evaluate the early results of the surgical treatment of active NVE and PVE. METHODS: During a 3-yr period (January 1 1996 December 31 1998), 57 out of 60 patients (pts) with active, either NVE (46 pts) or PVE (11 pts) underwent surgical treatment. There were 11 women (23.9%), average age of the group being 43.3+/-9.1yr (18-73). They were operated on 12 35days, mean 17.7+/-7.5days (for NVE) and 5-33days, mean 13.2+/-10.1days (for PVE) after the diagnosis of endocarditis was first suspected. All pts had at least one absolute indication for early surgical treatment, the most frequent being (in NVE) worsening heart failure (19 cases) and inability to control the infection (10 cases), while in PVE it was valve dehiscence (8 cases). In 8 cases of NVE and 2 cases of PVE fresh, antibiotic sterilized aortic homograft was used to replace the aortic valve. RESULTS: Operative mortality was 1.8% (1/57) and hospital mortality 5.2% (3/57). Three pts with PVE died before they were operated on, giving an overall mortality of 10% (6/60). Postoperative morbidity included valve dehiscence in two pts (probable late onset recurrent endocarditis - 3.5%), three episodes of acute renal failure (5.3%), four cases of respiratory insufficiency (7.0%) and one chronic pleural effusion (1.8%). All pts that were discharged from the hospital (54/60), are still alive and well 1-35months postoperatively (mean 20.3+/-9.6months), including pts with recurrent endocarditis and valve dehiscence, after they were successfully reoperated. CONCLUSIONS: Along with early diagnosis and appropriate antibiotic treatment, aggressive surgical attitude is of importance for the successful outcome in this group of seriously ill patients. Our data indicate that early surgical treatment in cases of active endocarditis may be associated with low mortality and morbidity. PMID- 10799832 TI - Celiac artery compression syndrome. AB - Celiac artery compression syndrome occurs when the median arcuate ligament of the diaphragm causes extrinsic compression of the celiac trunk. We report a case of a 65-year-old woman who presented with a three-month history of postprandial abdominal pain, nausea and some emesis, without weight loss. There was a bruit in the upper mid-epigastrium and the lateral aortic arteriography revealed a significant stenosis of the celiac artery. At operation, the celiac axis was found to be severely compressed anteriorly by fibers forming the inferior margin of the arcuate ligament of the diaphragm. The ligament was cut and a vein by-pass from the supraceliac aorta to the distal celiac artery was performed. The patient remains well and free of symptoms two and a half years since operation.In this report we discuss the indications and the therapeutic options of this syndrome as well as a review of the literature is being given. PMID- 10799831 TI - Trousseau's syndrome and acute arterial thrombosis. AB - This report describes three patients treated for acute arterial thrombosis due to malignancy-related hypercoagulability (Trousseau's syndrome). The average age was 59yr. There were two women and one man. The cancers were breast, lung, and pancreas. Atherosclerosis or nonneoplastic hypercoagulable states did not appear to be a factor in any patient. One patient who presented with irreversible arm ischemia and Stage IV breast cancer underwent primary amputation. The other two patients underwent immediate surgical thrombectomy and thrombolytic therapy, and malignancy was discovered during postoperative workup for hypercoagulable states. Both ultimately required amputation. All three patients died due to cancer less than one year after presentation. When a hypercoagulable state is suspected as the cause of acute arterial thrombosis, an evaluation for occult malignancy is indicated. Although aggressive revascularization attempts may be appropriate, the prognosis for limb salvage and long-term survival is poor. PMID- 10799833 TI - New targets, new drugs. PMID- 10799834 TI - Clinical studies in the development of new anticancer agents exhibiting growth inhibition in models: facing the challenge of a proper study design. AB - Various new specifically targeted anticancer agents such as matrixmetalloproteinase inhibitors, angiogenesis inhibitors, farnesyl transferase inhibitors, and tyrosine kinase inhibitors have been developed in recent years. These agents exert antitumor activity through specific target inhibition, and preclinical studies showed dose-dependent tumor growth inhibition and only sporadic tumor regressions. Toxicity of these new agents was often strikingly mild or did not occur at all. Clinical studies are now being performed. Because these new agents might have a different toxicity profile and exert their antitumor activity in a way that is completely different from that of cytotoxic agents, the design of clinical studies will have to be adapted in several ways, and new endpoints for phase I, II, and III studies must be defined. PMID- 10799835 TI - Target molecules for anti-angiogenic therapy: from basic research to clinical trials. AB - There is growing evidence that anti-angiogenic drugs will improve future therapies of diseases like cancer, rheumatoid arthritis and ocular neovascularisation. However, it is still uncertain which kind of substance, out of the large number of angiogenesis inhibitors, will prove to be a suitable agent to treat these human diseases. There are currently more than 30 angiogenesis inhibitors in clinical trials and a multitude of promising new candidates are under investigation in vitro and in animal models. Important therapeutic strategies are: suppression of activity of the major angiogenic regulators like vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF); inhibition of function of alphav-integrins and matrix metalloproteinases (MMPs); the exploitation of endogenous anti-angiogenic molecules like angiostatin, endostatin or thrombospondin. Given the wide spectrum of diseases which could be treated by anti-angiogenic compounds, it is important for today's clinicians to understand their essential mode of action at a cellular and molecular level. Here we give an in-depth overview of the basic pathophysiological mechanisms underlying the different anti-angiogenic approaches used to date based on the most recent fundamental and clinical research data. The angiogenesis inhibitors in clinical trials are presented and promising future drug candidates are discussed. PMID- 10799836 TI - Telomerase: a therapeutic target for the third millennium? AB - Telomerase offers the potential opportunity to control cell proliferation by interfering with a totally new and unique biological process which is cell senescence. The aim of this review is to impartially present the state of the art in telomerase with the pros and the cons of the current scientific situation of this fast-growing and fascinating topic for answering the key question asked by experimental and medical oncologists: Will telomerase be a therapeutic target for the third millenium? The most convincing argument (which is a scientifically documented one) for going ahead with this target is obviously the strong correlation existing between the level and frequency of telomerase expression and the malignant properties of tumors. This has been now largely documented in established tumor cell lines and fresh tumor samples obtained from patients. Noteworthy is the very important difference of telomerase expression between malignant and normal tissues. This difference is much higher than those observed for classical enzymatic targets of chemotherapy such as thymidylate synthetase, dihydrofolate reductase and topoisomerases. If this translates to the clinical situation, telomerase inhibitors might display a good selectivity for tumor cells with a minimal toxicity for normal tissues. The most appealing criticism (which is still purely speculative) is obviously the clinical relevance of inhibiting telomerase in cancer patients. According to the paradigm currently proposed for telomeres and telomerases, it can be predicted that telomerase inhibition will not affect a tumor until its telomeres reach the critical size for entering senescence. This means that during anti-telomerase therapy, the tumor cells will continue grow undergoing 20-30 divisions until the telomeres reach a critical size leading to tumor senescence. Does this make sense, especially in patients with advanced tumors at the beginning of the therapy? Ultimately, the definitive answer to the question will not come from intellectual speculation but from the properties of telomerase inhibitors, first in tumor bearing animals, then finally in cancer patients! Several institutions are very active in the development of telomerase inhibitors. Different stategies are used: direct inhibition of telomerase, interference with telomeres (G quartets), interaction with other proteins involved in the regulation of telomerase and telomeres. PMID- 10799837 TI - Staurosporine analogues - pharmacological toys or useful antitumour agents? AB - This review summarises the evidence for the potential antineoplastic activity of the staurosporine analogues 7-hydroxystaurospine (UCN-01) and N benzoylstaurosporine (CGP 41251) and defines the role of the enzyme family protein kinase C (PKC) in the mechanisms by which these agents interfere with malignant cell growth. PKC function is altered in some neoplasias, and this dysfunction has been related to uncontrolled proliferation. PKC also influences resistance of cancer cells against cytotoxic drugs. Staurosporine analogues compete with ATP, even though the exact action by which they inhibit PKC is more complicated. Staurosporine analogues do not exhibit specificity for particular PKC isoenzymes, but they inhibit 'conventional' PKC isoenzymes more potently than 'novel' and 'atypical' ones. They also interfere directly with the cell cycle machinery. Both CGP 41251 and UCN-01 are currently progressing through clinical evaluation. There is a remarkable difference in pharmacokinetic handling of CGP 41251 and UCN-01 between rodents and humans. CGP 41251 and UCN-01 might offer advantages in cancer therapy when applied in combination with conventional cytotoxic agents. PMID- 10799838 TI - Study of the combined influence of environmental factors on viability of cryptosporidium parvum oocysts in water evaluated by fluorogenic vital dyes and excystation techniques. AB - Using a factorial experimental design, the combined effect of salinity, temperature and storage time on the viability of Cryptosporidium parvum oocysts in water was evaluated by fluorogenic vital dyes (4',6-diamidino-2-phenylindole and propidium iodide) and an excystation technique. Salinity, storage time and their interaction seemed to be the most influential factors, whereas temperature was not a significant factor. Under unfavourable conditions (salinity 35 per thousand, storage time 40 days), even more than 20% of oocysts remain viable, indicating a high risk of infection for immunocompromised individuals. PMID- 10799839 TI - Viability and infectivity of two Cryptosporidium parvum bovine isolates from different geographical location. AB - The viability of two Cryptosporidium parvum bovine isolates from Spain and Colombia was evaluated by in vitro excystation, inclusion/exclusion of two fluorogenic vital dyes (DAPI and PI) and infectivity assay in a suckling murine model. Excystation percentages were similar for both Spain and Colombia isolates (83% and 87%, respectively). The total viability of the Spain isolate, measured by inclusion/exclusion of two fluorogenic vital dyes, was 71% in comparison with that detected for oocysts of the Colombia isolate, 32.3%. The bovine C. parvum oocysts of both isolates were viable and infectious for suckling Swiss CD-1 mice. However, infectivity percentage and the mean intensity of infection were consistently higher in the Spain isolate than those from Colombia isolate. It was not possible to obtain a good correlation between in vitro excystation, inclusion/exclusion of vital dyes and in vivo infectivity for the Colombia isolate, while data obtained with the Spain isolate indicated that there was an apparent strong correlation between excystation efficiency, total viability and the infectivity. Although a comparative analysis of genetic variation among these isolates from different geographical location is necessary, variations observed between the both isolates seemed to be a result of parasite adaptation to environmental stresses such as temperature which appears to have a direct effect on the permeability of the oocysts. PMID- 10799840 TI - Prevalence of Cryptosporidium in sheep and goats bred on five farms in west central region of Poland. AB - Faecal specimens were taken from 205 sheep and goats housed in five different localities in the west-central part of Poland. All faecal specimens were examined for Cryptosporidium by using microscopy screening of smears stained by modified Ziehl-Neelsen technique and commercial enzyme immunoassay. PCR technique using genus specific primers was additionally applied in the surveys of 10 faecal specimens collected from lambs. C. parvum infection was identified in 16 of 159 sheep (10.1%). Lambs were more often infected than adult sheep, and the intensity of infection was higher in lambs than in sheep, as a rule. Both lambs and sheep examined in the study were asymptomatically infected with Cryptosporidium. Both microscopy and enzyme immunoassay methods gave one false negative result. The examination of 10 faecal samples revealed 100% agreement among the results obtained by microscopic, immunologic and molecular methods. None of the goats raised on three farms were infected with Cryptosporidium. PMID- 10799841 TI - Control of gastrointestinal nematodes in first season grazing calves by two strategic treatments with eprinomectin. AB - A study was carried out to evaluate the effects of strategic early-season treatments with eprinomectin on first-season grazing calves exposed to strongyle infections on a naturally contaminated pasture. Two groups of first grazing season (FGS) calves were turned out in mid-May on two plots that were similar with respect to size and herbage infectivity. They grazed separately until housing at the end of October. One of these groups was given eprinomectin pour-on at turnout and 8 weeks later, while the other group served as untreated controls. The results showed that the treatments reduced gastrointestinal strongyle infections throughout the season as evidenced by lower faecal egg counts and serum pepsinogen levels compared with the controls. Furthermore, the results of herbage larval counts and postmortem worm counts in tracer animals demonstrated that the treatment had reduced herbage infectivity on the 'treated' plot. Finally, the chemoprophylactically treated calves had a better weight gain over the duration of the study than the untreated controls. PMID- 10799842 TI - Nematode parasites of adult dairy cattle in the Netherlands. AB - Abomasa, blood samples and faecal samples for examination of nematode infections were collected from 125 dairy cows during the period November 1997-October 1998. Of these, 12 had no grazing history and were, therefore, excluded from this study. From the remaining 113, 88.5% had nematode eggs in the faeces. Larval identification of the positive cultures showed that Ostertagia spp. larvae were most frequent (97%), followed by Trichostrongylus spp. (29%), Oesophagostomum spp. (23%), Cooperia punctata (20%), Cooperia oncophora (4%), Haemonchus contortus (2%) and Bunostomum phlebotomum (1%). The geometric mean EPG was 2.4. Two cows excreted larvae of Dictyocaulus viviparus (0.1 and 0.6 LPG resp.). Worms were found in the abomasa of 108 cows (96%). In all these abomasa Ostertagia spp. was present (100%). Trichostrongylus axei was found in 47 abomasa (43.5%) and two cows (2%) were infected with Capillaria bovis. The geometric mean of the total abomasal worm counts was 1743 and of Ostertagia spp. alone 1615. Almost all male worms were Ostertagia ostertagi, only occasionally Skrjabinagia lyrata10,000) total worm burden. Ostertagia specific antibodies were highest in late summer and autumn and lowest in spring and early summer. The same pattern, although not so pronounced, was observed for the serum pepsinogen values. No clear seasonal pattern was found for the Cooperia specific antibodies. Antibodies against D. viviparus were detected in seven cows (6%). PMID- 10799843 TI - Resistance of Holstein-Friesian cows to infestation by the cattle tick (Boophilus microplus). AB - In two experiments, the milk production of 59 Holstein-Friesian cows in mid lactation was measured over 1 week before artificial infestation with 2,500 or 5,000 Boophilus microplus larvae. Host resistance, the proportion of female ticks applied but not engorging was estimated from weekly counts of engorging female ticks of 4.5-8. 0mm long. Mean host resistance was 79 and 67% in two experiments. Host resistance was not significantly related to milk yield before infestation, to early pregnancy, or to parity. Culling the 10% of cattle with the least resistance to ticks would result in removal of 19-21% of ticks in a herd. The results suggest that selection to improve the existing low levels of resistance to B. microplus might be undertaken without compromising milk production. However, low levels of resistance among the cattle studied and difficulties in assessment of resistance are likely to limit the usefulness of selection within the Holstein-Friesian breed. PMID- 10799844 TI - Activity of decoquinate against Cryptosporidium parvum in cell cultures and neonatal mice. AB - Cryptosporidium parvum is an apicomplexan parasite that is an important cause of diarrhea in neonatal calves and humans. No treatment is currently available for neonatal calves. We have recently learned from colleagues in the pharmaceutical industry that dairy practitioners are sometimes using decoquinate for the treatment of neonatal bovine cryptosporidiosis. Therefore, the present study was undertaken to determine whether the clinical observations in calves can be substantiated by laboratory investigation. Oocysts of the KSU-1 isolate of C. parvum were used to infect human ileocecal epithelial cells in vitro to measure the efficacy of treatment using an ELISA based assay. No activity was observed at 10 or 50microM decoquinate, but at 100microM an 8% inhibition of development was seen. Oocysts of the AUCp-1 isolate of C. parvum were then used to infect suckling mice. The numbers of oocysts observed in suckling mice treated with 2.5 or 5.0mg/kg decoquinate were not significantly different from untreated control suckling mice (p0.05). The results of our study suggest that decoquinate should have little efficacy for treatment of neonatal bovine cryptosporidiosis if administered once per day and that any clinical improvement observed in treated calves may be due to factors unrelated to decoquinate's effect on C. parvum. PMID- 10799845 TI - Influence of in ovo administered Cryptosporidium baileyi oocyst extract on the course of homologous infection. AB - In order to evaluate the effect of in ovo vaccination on avian cryptosporidiosis, two doses (1 and 10microg) of Cryptosporidium baileyi oocyst extract (OE) were injected into the amnionic sac of embryonated, specific pathogen-free chicken eggs. After hatching these birds as well as infected controls (IC) were inoculated with 8x10(5) C. baileyi oocysts at 10 days of age. Another group of chickens remained uninfected (UC). Faecal oocyst shedding was measured every second day, and weekly ELISAs were performed to monitor seroconversion. Those chickens that received OE during embryogenesis showed dose-dependent shift in their oocyst shedding, with higher oocyst output of OE1 and OE10 birds compared to IC ones. The patency was significantly longer in the OE10 group than in IC or OE1. ELISA results showed low seroconversion of OE1 and OE10 chickens prior to homologous challenge. Challenge infection resulted in antibody levels without significant difference between IC, OE1 and OE10 groups. These data suggest that in ovo vaccination with C. baileyi oocyst extract does not promote immune response, moreover, it may impair immunity and thus delay the clearance of cryptosporidia from chickens. PMID- 10799846 TI - Cross-reactivity of anti-Taenia crassiceps cysticerci immune antibodies with Taenia solium antigens. AB - A comparative study of antibody production was carried out using BALB/c mice immunized with 20 or 50microg vesicular fluid (VF)-Tcra (Taenia crassiceps) antigens, and gel of <30kD or eluate from <30kD peptides. Good IgM, IgA and IgG levels were detected by ELISA-Tcra and the antibodies presented reactivity with the <20kD peptides when tested by immunoblotting-Tcra. The antibodies from animals immunized with 20 and 50microg presented high anti-Tso cross-reactivity in ELISA (IgG>>IgM and IgA). All groups presented IgG antibodies identifying the 12kD Tso-peptide. PMID- 10799847 TI - Seasonal patterns of strongyle infections in grazing sheep under the traditional production system in the region of Trikala, Greece. AB - Ten 4-month-old female sheep of the Karagouniko dairy breed were used to monitor the seasonal fluctuations of strongyle infections in sheep during the first year of grazing under the traditional production system in the region of Trikala, Greece, where control of nematodes is currently based exclusively on the frequent use of anthelmintics which might contribute to the appearance of anthelmintic resistance. The sheep grazed communal pasture plots and did not receive any anthelmintic treatment during the entire study period. Faecal egg counts (epg), plasma pepsinogen levels, blood values, and genera of parasitic strongyles recovered from coprocultures were recorded monthly. Mean epg for strongyle-type eggs were significantly higher during summer. Mean plasma pepsinogen levels were significantly higher during spring and summer. The genera of parasitic strongyles recovered from the faecal cultures were Haemonchus, Ostertagia, Trichostrongylus, Cooperia, Nematodirus, Oesophagostomum, and Chabertia. The percentages of larvae for Haemonchus, Ostertagia, Cooperia, and Oesophagostomum were significantly higher during winter. The percentages of larvae for Trichostrongylus were significantly higher during summer. During summer, levels of RBC, HGB, and HCT were depressed and levels of MCV, MCH, and MCHC were elevated. Mean epg for strongyle-type eggs had a significant inverse correlation with RBC, HGB, HCT, and positive correlation with MCH, MCHC, and percentages of eosinophils in differential leucocyte counts. The seasonal pattern of infection observed in the present study indicates that it is possible to decrease the number of treatments to one per year, thus, reducing the possibility for the appearance of anthelmintic resistance. PMID- 10799849 TI - Cutting edge: inflammatory responses can be triggered by TREM-1, a novel receptor expressed on neutrophils and monocytes. AB - We have identified new activating receptors of the Ig superfamily expressed on human myeloid cells, called TREM (triggering receptor expressed on myeloid cells). TREM-1 is selectively expressed on blood neutrophils and a subset of monocytes and is up-regulated by bacterial LPS. Engagement of TREM-1 triggers secretion of IL-8, monocyte chemotactic protein-1, and TNF-alpha and induces neutrophil degranulation. Intracellularly, TREM-1 induces Ca2+ mobilization and tyrosine phosphorylation of extracellular signal-related kinase 1 (ERK1), ERK2 and phospholipase C-gamma. To mediate activation, TREM-1 associates with the transmembrane adapter molecule DAP12. Thus, TREM-1 mediates activation of neutrophil and monocytes, and may have a predominant role in inflammatory responses. PMID- 10799850 TI - Cutting edge: differential constitutive expression of functional receptors for lysophosphatidic acid by human blood lymphocytes. AB - Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) from platelets and macrophages mediate T cell functions. Endothelial differentiation gene-encoded G protein-coupled receptors (Edg Rs) are specific for S1P (Edg-1, -3, -5, and -8 Rs) and LPA (Edg-2, -4, and -7 Rs). Human T cell tumors express many Edg Rs for both LPA and S1P. In contrast, human blood CD4+ T cells express predominantly Edg 4, and CD8+ T cells show only traces of Edg-2 and -5, by quantification of mRNA and Edg R Ags. LPA at 10-10-10-6 M suppressed significantly the secretion of IL-2 from anti-CD3 plus anti-CD28 Ab-challenged CD4+ T cells, but not CD8+ T cells. Monoclonal anti-Edg-4 R Ab, like LPA, suppressed stimulated IL-2 secretion from CD4+ T cells, but not CD8+ T cells. Constitutive expression of Edg-4 by CD4+, but not CD8+, human T cells accounts for differential functional responsiveness of the T cell subsets to LPA. PMID- 10799851 TI - Cutting edge: a role for CD1 in the pathogenesis of lupus in NZB/NZW mice. AB - Since anti-CD1 TCR transgenic T cells can activate syngeneic B cells via CD1 to secrete IgM and IgG and induce lupus in BALB/c mice, we studied the role of CD1 in the pathogenesis of lupus in NZB/NZW mice. Approximately 20% of B cells from the spleens of NZB/NZW mice expressed high levels of CD1 (CD1high B cells). The latter subset spontaneously produced large amounts of IgM anti-dsDNA Abs in vitro that was up to 25-fold higher than that of residual CD1int/low B cells. T cells in the NZB/NZW spleen proliferated vigorously to the CD1-transfected A20 B cell line, but not to the parent line. Treatment of NZB/NZW mice with anti-CD1 mAbs ameliorated the development of lupus. These results suggest that the CD1high B cells and their progeny are a major source of autoantibody production, and activation of B cells via CD1 may play an important role in the pathogenesis of lupus. PMID- 10799852 TI - Cutting edge: the IgG response to the circumsporozoite protein is MHC class II dependent and CD1d-independent: exploring the role of GPIs in NK T cell activation and antimalarial responses. AB - Biochemical analysis has suggested that self GPI anchors are the main natural ligand associated with mouse CD1d molecules. A recent study reported that Valpha14+ NK T cells responded to self as well as foreign (parasite-derived) GPIs in a CD1d-dependent manner. It further reported that the IgG response to the Plasmodium berghei malarial circumsporozoite (CS) protein was severely impaired in CD1d-deficient mice, leading to a model whereby NK T cells, upon recognition of CD1d molecules presenting the CS-derived GPI anchor, provide help for B cells secreting anti-CS Abs. We tested this model by comparing the anti-CS Ab responses of wild-type, CD1d-deficient, and MHC class II-deficient mice. We found that the IgG response to the CS protein was solely MHC class II-dependent. Furthermore, by measuring the response of a broad panel of CD1d-autoreactive T cells to GPI deficient CD1d-expressing cells, we found that GPIs were not required for autoreactive responses. PMID- 10799853 TI - Cutting edge: stromal cell-derived factor-1 is a costimulator for CD4+ T cell activation. AB - Stromal cell-derived factor (SDF)-1 is a chemoattractant for T cells, precursor B cells, monocytes, and neutrophils. SDF-1alpha was also found to up-regulate expression of early activation markers (CD69, CD25, and CD154) by anti-CD3 activated CD4+ T cells. In addition, SDF-1alpha costimulated proliferation of CD4+ T cells and production of IL-2, IFN-gamma, IL-4, and IL-10. Stimulation with SDF-1alpha alone did not induce activation marker expression, proliferation, or cytokine production by the CD4+ T cells. SDF-1alpha-mediated costimulation was blocked by anti-CXC chemokine receptor-4 mAb. RANTES also increased activation marker expression by anti-CD3-stimulated peripheral CD4+ T cells, but less effectively than SDF-1alpha did, and did not up-regulate IL-2 production and proliferation. These results indicate that SDF-1 and CXC chemokine receptor-4 interactions not only play a role in T cell migration but also provide potent costimulatory signals to Ag-stimulated T cells. PMID- 10799854 TI - Cutting edge: a soluble form of CTLA-4 in patients with autoimmune thyroid disease. AB - We have recently identified a novel transcript of the CTLA-4 gene that may represent a native soluble form of CTLA-4 (sCTLA-4). To determine whether sCTLA-4 was expressed in humans, we applied a sensitive enzyme immunoassay on serum from patients with autoimmune thyroid disease (ATD). Eleven of 20 patients with ATD had circulating levels of sCTLA-4 ranging from 28 to 78 ng/ml, whereas only 1 of 30 apparently healthy volunteers had a level greater than 4 ng/ml. sCTLA-4 immunoreactivity was inhibited by its binding to B7.1, suggesting that sCTLA-4 is a functional receptor. Immunoprecipitation analysis of serum from patients with ATD revealed a polypeptide consistent with the predicted size of sCTLA-4. We conclude that a native soluble form of CTLA-4 is derived from an alternate transcript of the CTLA-4 gene, and its level in plasma is elevated among a population of patients with ATD. PMID- 10799855 TI - Cutting edge: the human cytomegalovirus UL40 gene product contains a ligand for HLA-E and prevents NK cell-mediated lysis. AB - Human CMV has evolved multiple strategies to interfere with immune recognition of the host. A variety of mechanisms target Ag presentation by MHC class I molecules resulting in a reduced class I cell-surface expression. This down-regulation of class I molecules is expected to trigger NK cytotoxicity, which would have to be counteracted by the virus to establish long-term infection. Here we describe that the human CMV open reading frame UL40 encodes a canonical ligand for HLA-E, identical with the HLA-Cw03 signal sequence-derived peptide. Expression of UL40 in HLA-E-positive target cells conferred resistance to NK cell lysis via the CD94/NKG2A receptor. Generation of the UL40-derived HLA-E ligand was also observed in TAP-deficient cells. The presence of a functional TAP-independent HLA E ligand in the UL40 signal sequence implicates this viral gene as an important negative regulator of NK activity. PMID- 10799856 TI - Cutting edge: the tumor counterattack hypothesis revisited: colon cancer cells do not induce T cell apoptosis via the Fas (CD95, APO-1) pathway. AB - The counterattack hypothesis, suggesting that cancer cells express Fas ligand (FasL) and are able to kill Fas-expressing tumor-infiltrating activated T cells, was supported by reports of the killing of Jurkat cells by FasL-expressing human colon cancer cell lines. Through the use of an improved cytotoxic assay in which soluble FasL and FasL-transfected KFL9 cells were used as positive controls, we show that none of seven human colon cancer cell lines induce apoptosis of two Fas expressing target cell lines, Jurkat and L1210-Fas cells. Moreover, in coculture experiments, cancer cell monolayers do not inhibit the growth of Fas-expressing lymphoid cells. Although FasL mRNA and protein were detected in the extracts of the colon cancer cell lines, flow cytometry and confocal microscopy failed to detect the protein on the surface of tumor cells. These results suggest that the counterattack of tumor-infiltrating T lymphocytes by cancer cells may not account for immune tolerance toward tumor cells. PMID- 10799857 TI - Human intestinal epithelial cells express a novel receptor for IgA. AB - Binding and transport of polymeric Igs (pIgA and IgM) across epithelia is mediated by the polymeric Ig receptor (pIgR), which is expressed on the basolateral surface of secretory epithelial cells. Although an Fc receptor for IgA (FcalphaR) has been identified on myeloid cells and some cultured mesangial cells, the expression of an FcalphaR on epithelial cells has not been described. In this study, binding of IgA to a human epithelial line, HT-29/19A, with features of differentiated colonic epithelial cells, was examined. Radiolabeled monomeric IgA (mIgA) showed a dose-dependent, saturable, and cation-independent binding to confluent monolayers of HT-29/19A cells. Excess of unlabeled mIgA, but not IgG or IgM, competed for the mIgA binding, indicating that the binding was IgA isotype-specific and was not mediated by the pIgR. The lack of competition by asialoorosomucoid and the lack of requirement for divalent cations excluded the possibility that IgA binding to HT-29/19A cells was due to the asialoglycoprotein receptor or beta-1, 4-galactosyltransferase, previously described on HT-29 cells. Moreover, the FcalphaR (CD89) protein and message were undetectable in HT-29/19A cells. FACS analysis of IgA binding demonstrated two discrete populations of HT 29/19 cells, which bound different amounts of mIgA. IgA binding to other colon carcinoma cell lines was also demonstrated by FACS analysis, suggesting that an IgA receptor, distinct from the pIgR, asialoglycoprotein receptor, galactosyltransferase, and CD89 is constitutively expressed on cultured human enterocytes. The function of this novel IgA receptor in mucosal immunity remains to be elucidated. PMID- 10799858 TI - Effects of continuous exposure to stromal cell-derived factor-1 alpha on T cell rolling and tight adhesion to monolayers of activated endothelial cells. AB - Immobilized stromal cell-derived factor-1 alpha (SDF-1 alpha) has been shown to induce tight adhesion of T cells to purified ICAM-1 in assays done under flow conditions. In this study, we show that soluble SDF-1 alpha induced a rapid (within 20 s) cessation of rolling and tight adhesion of >90% of the rolling T cells on monolayers of activated endothelial cells under similar flow. Within 4 min, the T cells had either started to migrate between the endothelial cells or re-entered the rolling and circulating lymphocyte pool. This deadherence of the firmly bound cells, with either ensuing transmigration or continued rolling, was most likely due to desensitization of lymphocytes to the continuously present SDF 1 alpha. The released rolling lymphocytes could still respond to other activating signals by a second round of tight adhesion. Pretreating the lymphocytes with pertussis toxin almost completely blocked the effect of the chemokine, confirming that the induction of firm adhesion was due to the function of the chemokine on the lymphocytes and not the endothelial cells. Pretreating the endothelium with SDF-1 alpha did not lead to firm adhesion of subsequently added lymphocytes, also indicating that the effect was due to soluble, not endothelially bound, chemokine. Blocking experiments showed that the same molecules mediated rolling before and after SDF-1 alpha-induced tight adhesion. This is the first study to demonstrate the effect of soluble SDF-1 alpha on T cell rolling on an endothelial cell monolayer. The data broaden our understanding of the stimulatory factors directing the firm adhesion and ensuing transmigration of leukocytes into tissues through activated endothelium. PMID- 10799859 TI - Perinatal deletion of B cells expressing surface Ig molecules that lack V(D)J encoded determinants in the bursa of Fabricius is not due to intrafollicular competition. AB - During embryonic development, the avian bursa of Fabricius selects B cell precursors that have undergone productive V(D)J recombination for expansion in oligoclonal follicles. During this expansion, Ig diversity is generated by gene conversion. We have used retroviral gene transfer in vivo to introduce surface Ig molecules that lack V(D)J-encoded determinants into B cell precursors. This truncated mu heavy chain supports both B cell expansion within embryo bursal lymphoid follicles and gene conversion. We show that individual follicles can be colonized exclusively by cells expressing the truncated mu chain and lacking endogenous surface IgM, ruling out a requirement for V(D)J-encoded determinants in the establishment of bursal lymphoid follicles. In striking contrast to their normal development in the embryo, bursal cells expressing the truncated mu-chain exhibit reduced rates of cell division and increased levels of apoptosis after hatching. The level of apoptosis in individual follicles reflects the proportion of cells within the follicle that express the truncated mu-chain. In particular, high levels of apoptosis are associated with follicles containing exclusively cells expressing the truncated micro receptor. Thus, apoptotic elimination of such cells is not due to competition within the follicle by cells expressing endogenous surface IgM receptors. This provides the first direct demonstration that the regulation of B cell development in the avian bursa after hatching differs fundamentally from that seen in the embryo. The requirement for intact IgM expression when the bursa is exposed to exogenous Ag implicates a role for Ag in avian B cell development after hatching. PMID- 10799860 TI - Quantification of cell turnover kinetics using 5-bromo-2'-deoxyuridine. AB - 5-Bromo-2'-deoxyuridine (BrdU) is frequently used to measure the turnover of cell populations in vivo. However, due to a lack of detailed mathematical models that describe the uptake and loss of BrdU in dividing cell populations, assessments of cell turnover kinetics have been largely qualitative rather than quantitative. In this study, we develop a mathematical framework for the analysis of BrdU-labeling experiments. We derive analytical expressions for the fraction of labeled cells within cell populations that are growing, declining, or at equilibrium. Fitting the analytical functions to data allows us to quantify the rates of cell proliferation and cell loss, as well as the rate of cell input from a source. We illustrate this for the BrdU labeling of T lymphocytes of uninfected and SIV infected rhesus macaques. PMID- 10799861 TI - CD40 engagement on synovial fibroblast up-regulates production of vascular endothelial growth factor. AB - We tested the impact of CD40 engagement on the production of vascular endothelial growth factor (VEGF) from rheumatoid synovial fibroblasts. Fibroblast-like synovial cells (FLS) were prepared from the synovial tissues of rheumatoid arthritis patients and cultured in the presence of CD40 ligand-transfected (CD40L+) L cells. VEGF levels were determined in the culture supernatants by ELISA. Stimulation of FLS by CD40L+ L cells increased the production of VEGF by 4.1-fold over the constitutive levels of unstimulated FLS. The CD40L on activated T cells from rheumatoid synovial fluid also up-regulated VEGF production from FLS. Neither indomethacin nor Abs to IL-1beta, TNF-alpha, and TGF-beta did affect CD40L-induced VEGF production. Stimulation of FLS with TNF-alpha, IL-1beta, and TGF-beta increased VEGF production by 1.6-, 2.0-, and 5.2-fold, respectively, and displayed an additive effect on the production of VEGF by CD40L. VEGF mRNA expression was also up-regulated by the stimulation of FLS with membranes from the CD40L+ L cells. Dexamethasone completely abrogated CD40L-induced VEGF production. In addition, pyrrolidine dithiocarbamate partially down-regulated CD40L-induced VEGF production, showing that the NF-kappaB pathway was partly involved in the signaling of CD40L leading to VEGF production. Collectively, these results suggest that the interaction between CD40 on synovial fibroblasts and CD40L expressed on activated T lymphocytes may be directly involved in the neovascularization in rheumatoid synovitis by enhancing the production of VEGF. PMID- 10799862 TI - Bioactivities of Fas ligand-expressing retroviral particles. AB - Culture supernatants from retroviral packaging cells carrying the human Fas ligand (FasL) gene killed both human (Jurkat) and mouse (LB27.4) targets within 5 h of incubation. Cytotoxicity was found both in a fraction >/=500 kDa and a fraction between 50 and 500 kDa. Following ultracentrifugation, the activity in the >/=500-kDa fraction was concentrated in the pellet (FasL vector preparation (VP)), which was also infective when added to NIH-3T3 cells. Both Polybrene and poly-l -lysine significantly enhanced the cytotoxicity of FasL VP but not anti Fas mAb, soluble FasL (sFasL), and cell-associated FasL. In the presence of Polybrene, FasL VP killed targets that are resistant to anti-Fas mAb and sFasL. The infectivity but not FasL cytotoxicity of FasL VP was sensitive to irradiation and heat shock. By contrast, cytotoxicity of FasL VP could be enhanced or inhibited depending on the doses of anti-FasL mAb. Interestingly, the infectivity of FasL VP was specifically enhanced by anti-FasL mAb, suggesting that a nonviral gene product could be used to regulate the behavior of the retroviral vector. Thus, in addition to expressing potent FasL cytotoxicity, the FasL VP exhibits unique properties heretofore not attributed to anti-Fas mAb, sFasL, and cell associated FasL. Our study raises the possibility of using the retroviral gene packaging technology to make powerful, versatile, and regulatable bioactive vesicles expressing a predetermined function of the protein encoded by the target gene. PMID- 10799864 TI - Engagement of CD4 before TCR triggering regulates both Bax- and Fas (CD95) mediated apoptosis. AB - In the present study, we have aimed at clarifying the CD4-dependent molecular mechanisms that regulate human memory T cell susceptibility to both Fas (CD95) dependent and Bcl-2-dependent apoptotic pathways following antigenic challenge. To address this issue, we used an experimental system of viral and alloantigen specific T cell lines and clones and two ligands of CD4 molecules, Leu-3a mAb and HIV gp120. We demonstrate that CD4 engagement before TCR triggering suppresses the TCR-mediated neosynthesis of the Flice-like inhibitory protein and transforms memory T cells from a CD95-resistant to a CD95-susceptible phenotype. Moreover, evidence that the apoptotic programs were executed while Fas ligand mRNA expression was inhibited led us to analyze Bcl-2-dependent pathways. The data show that the engagement of CD4 separately from TCR influences the expression of the proapoptotic protein Bax independently of the anti-apoptotic protein Bcl-2, whereas Ag activation coordinately modulates both Bax and Bcl-2. The increased expression of Bax and the consequent dissipation of the mitochondrial transmembrane potential (DeltaPsim) suggest a novel immunoregulatory function of CD4 and demonstrate that both passive cell death and activation-induced cell death are operative in CD4+ memory T cells. Furthermore, analysis of the mechanisms by which IL-2 and IL-4 cytokines exert their protective function on CD4+ T cells in the presence of soluble CD4 ligands shows that they were able to revert susceptibility to Bax-mediated but not to CD95-dependent apoptotic pathways. PMID- 10799863 TI - Sequential cleavage by metallopeptidases and proteasomes is involved in processing HIV-1 ENV epitope for endogenous MHC class I antigen presentation. AB - Antigenic peptides derived from viral proteins by multiple proteolytic cleavages are bound by MHC class I molecules and recognized by CTL. Processing predominantly takes place in the cytosol of infected cells by the action of proteasomes. To identify other proteases involved in the endogenous generation of viral epitopes, specifically those derived from proteins routed to the secretory pathway, we investigated presentation of the HIV-1 ENV 10-mer epitope 318RGPGRAFVTI327 (p18) to specific CTL in the presence of diverse protease inhibitors. Both metalloproteinase and proteasome inhibitors decreased CTL recognition of the p18 epitope expressed from either native gp160 or from a chimera based on the hepatitis B virus secretory core protein as carrier protein. Processing of this epitope from both native ENV and the hepatitis B virus secretory core chimeric protein appeared to proceed by a TAP-dependent pathway that involved sequential cleavage by proteasomes and metallo-endopeptidases; however, other protease activities could replace the function of the lactacystin sensitive proteasomes. By contrast, in a second TAP-independent pathway we detected no contribution of metallopeptidases for processing the ENV epitope from the chimeric protein. These results show that, in the classical TAP-dependent MHC class I pathway, endogenous Ag processing of viral proteins to yield the p18 10 mer epitope requires metallo-endopeptidases in addition to proteasomes. PMID- 10799865 TI - Kappa-opioid regulation of thymocyte IL-7 receptor and C-C chemokine receptor 2 expression. AB - Endogenous and exogenous kappa-opioid agonists have been widely reported to modulate the immune response. We have published results that show that the superantigen-induced proliferative response of thymocytes is inhibited by the selective kappa-opioid agonist trans-3, 4-dichloro-N-methyl-N-[2-(1 pyrolidinyl)cyclohexyl] benzeneaceamide methanesulfonate (U50,488H). Previous work has established that the kappa-opioid receptor is widely expressed within the thymus; however, little is known about the role of the kappa-opioid receptor in the function of thymocytes. In the present report, we have examined the impact of U50,488H administration on the expression of cytokines in superantigen stimulated thymocytes by RNase protection analysis. We have measured detectable levels of the cytokines IL-2, IL-4, IL-5, IL-13, and IFN-gamma, and the chemokines lymphotactin and RANTES, in stimulated thymocyte cultures; however, addition of U50,488H did not alter the expression of these cytokines. Examination of cytokine receptor expression by these thymocytes revealed a significant inhibition in the expression of the transcript for the IL-7 receptor alpha-chain (IL-7Ralpha), and these results were confirmed by flow cytometry. Surprisingly, the expression of several other cytokine receptor chains including the common gamma-chain, IL-2Rbeta, or the IL-2Ralpha, IL-4Ralpha, and IL-15Ralpha chains, was not altered. In contrast to these results, a significant elevation in the expression of the chemokine receptor CCR2 was observed in U50,488H-treated cultures. These results suggest that the kappa-opioid receptor may function to promote cellular migration at the expense of the sensitivity to the growth promoting/maturation activity of IL-7. PMID- 10799867 TI - Phagosomes acquire nascent and recycling class II MHC molecules but primarily use nascent molecules in phagocytic antigen processing. AB - Phagosomes contain class II MHC (MHC-II) and form peptide:MHC-II complexes, but the source of phagosomal MHC-II molecules is uncertain. Phagosomes may acquire nascent MHC-II or preexisting, recycling MHC-II that may be internalized from the plasma membrane. Brefeldin A (BFA) was used to deplete nascent MHC-II in murine macrophages to determine the relative contributions of nascent and recycling MHC II molecules to phagocytic Ag processing. In addition, biotinylation of cell surface proteins was used to assess the transport of MHC-II from the cell surface to phagosomes. BFA inhibited macrophage processing of latex bead-conjugated Ag for presentation to T cells, suggesting that nascent MHC-II molecules are important in phagocytic Ag processing. Furthermore, detection of specific peptide:MHC-II complexes in isolated phagosomes confirmed that BFA decreased formation of peptide:MHC-II complexes within phagosomes. Both flow organellometry and Western blot analysis of purified phagosomes showed that about two-thirds of phagosomal MHC-II was nascent (depleted by 3 h prior treatment with BFA) and primarily derived from intracellular sites. About one-third of phagosomal MHC-II was preexisting and primarily derived from the plasma membrane. BFA had little effect on phagosomal H2-DM or the degradation of bead-associated Ag. Thus, inhibition of phagocytic Ag processing by BFA correlated with depletion of nascent MHC-II in phagosomes and occurred despite the persistent delivery of plasma membrane-derived recycling MHC-II molecules and other Ag-processing components to phagosomes. These observations suggest that phagosomal Ag processing depends primarily on nascent MHC-II molecules delivered from intracellular sites, e.g., endocytic compartments. PMID- 10799866 TI - The mouse tumor cell lines EL4 and RMA display mosaic expression of NK-related and certain other surface molecules and appear to have a common origin. AB - As a potential means for facilitating studies of NK cell-related molecules, we examined the expression of these molecules on a range of mouse tumor cell lines. Of the lines we initially examined, only EL4 and RMA expressed such molecules, both lines expressing several members of the Ly49 and NKRP1 families. Unexpectedly, several of the NK-related molecules, together with certain other molecules including CD2, CD3, CD4, CD32, and CD44, were often expressed in a mosaic manner, even on freshly derived clones, indicating frequent switching in expression. In each case examined, switching was controlled at the mRNA level, with expression of CD3zeta determining expression of the entire CD3-TCR complex. Each of the variable molecules was expressed independently, with the exception that CD3 was restricted to cells that also expressed CD2. Treatment with drugs that affect DNA methylation and histone acetylation could augment the expression of at least some of the variable molecules. The striking phenotypic similarity between EL4 and RMA led us to examine the state of their TCRbeta genes. Both lines had identical rearrangements on both chromosomes, indicating that RMA is in fact a subline of EL4. Overall, these findings suggest that EL4 is an NK-T cell tumor that may have retained a genetic mechanism that permits the variable expression of a restricted group of molecules involved in recognition and signaling. PMID- 10799868 TI - Granulocyte/macrophage colony-stimulating factor inhibits IL-12 production of mouse Langerhans cells. AB - We investigated the capacity of mouse Langerhans cells (LC) to produce IL-12, a central cytokine in a Th1 type of immune responses. We prepared purified LC (>95%) from BALB/c mouse skin by the panning method using anti-I-Ad mAb. An ELISA showed that purified LC spontaneously produced IL-12 p40, and that its production was up-regulated following simultaneous stimulation with anti-CD40 mAb and IFN gamma. Surprisingly, GM-CSF strikingly inhibited IL-12 p40 production by anti CD40/IFN-gamma-stimulated LC (% inhibition = 97.0 +/- 0.9% at 1 ng/ml GM-CSF). Supernatants of 48-h cultured keratinocytes (KC) also caused the inhibition of LC IL-12 p40 secretion, and this effect was neutralized by anti-GM-CSF mAb. IL 1alpha (1 ng/ml)-stimulated KC produced much more GM-CSF than unstimulated KC (60.9 +/- 0.2 pg/ml vs 20.9 +/- 1.7 pg/ml), and IL-1alpha-stimulated KC supernatants strongly inhibited IL-12 p40 production by anti-CD40/IFN-gamma stimulated LC (% inhibition = 89.4 +/- 1.4%). A bioassay using an IL-12-dependent T cell line demonstrated the correlation of the level of IL-12 p40 with the bioactivity of IL-12. These results provide important implications for the pathogenesis of atopic dermatitis, which involves the participation of LC and KC with the capacity to produce IL-12 and GM-CSF, respectively. PMID- 10799870 TI - Systemic immune deviation in the brain that does not depend on the integrity of the blood-brain barrier. AB - OVA injected into the brain of normal mice evoked a deviant immune response (brain-associated immune deviation (BRAID)) that was deficient in OVA-specific delayed-type hypersensitivity. This response was not dependent on an intact blood brain barrier since BRAID was induced even when OVA was injected into a newly created lesion site with extensive BBB leakage. However, newly activated microglia at the injection site 2 days after ablation of the striatum correlated with the loss of BRAID. At day 4 after trauma, when activated microglia were only visible further away from the injection site, BRAID was again able to be induced. In contrast to immune deviation elicited via the eye, an intact spleen was not required for BRAID, nor was BRAID adoptively transferable with spleen cells. In contrast i.v. injection of cervical lymph node cells harvested 8 days after OVA injection into the striatum was able to transfer BRAID into naive animals. Together, these data indicate that immune privilege in the brain is actively maintained and is mediated by an immune deviation mechanism that differs from eye derived immune deviation and arises even when the BBB is compromised. PMID- 10799869 TI - Potassium regulates IL-1 beta processing via calcium-independent phospholipase A2. AB - We report that potassium leakage from cells leads to activation of the Ca2+ independent phospholipase A2 (iPLA2), and the latter plays a pivotal role in regulating the cleavage of pro-IL-1 beta by the IL-converting enzyme caspase-1 in human monocytes. K+ efflux led to increases of cellular levels of glycerophosphocholine, an unambiguous indicator of phospholipase A2 activation. Both maturation of IL-1 beta and formation of glycerophosphocholine were blocked by bromoenol lactone, the specific iPLA2 inhibitor. Bromoenol lactone-dependent inhibition of IL-1 beta processing was not due to perturbation of the export machinery for pro-IL-1 beta and IL-1 beta or to caspase-1 suppression. Conspicuously, activation of Ca2+-dependent phospholipase A2 did not support but rather suppressed IL-1 beta processing. Thus, our findings reveal a specific role for iPLA2 activation in the sequence of events underlying IL-1 beta maturation. PMID- 10799871 TI - Mechanisms of graft acceptance: evidence that plasminogen activator controls donor-reactive delayed-type hypersensitivity responses in cardiac allograft acceptor mice. AB - We have used delayed-type hypersensitivity (DTH) responses to probe the mechanisms of drug-induced cardiac allograft acceptance in mice. DBA/2-->C57BL/6 cardiac allograft recipients treated transiently with gallium nitrate accept their grafts for >90 days and fail to display DBA/2-reactive DTH responses. These DTH responses are restored when anti-TGF-beta Abs are included at the challenge site, and cell depletion studies showed that this DTH inhibition is mediated by CD4+ cells. Real-time PCR analysis revealed that allograft acceptor mice produce no more than background levels of TGF-beta mRNA at DTH challenge sites. This suggests that DTH regulation in allograft acceptor mice may involve TGF-beta activation, rather than TGF-beta production. The protease, plasmin, can activate TGF-beta, and activated T cells can express a receptor for the plasmin-producing enzyme urokinase-type plasminogen activator (uPA), and can also produce both uPA and tissue-type plasminogen activator (tPA). We observed that Abs to tPA or uPA can replace anti-TGF-beta mAb for the restoration of donor-reactive DTH responses in allograft acceptor mice. Histologic analysis revealed that accepted cardiac allografts express uPA, tPA, and active TGF-beta, whereas accepted cardiac isografts express only tPA, but not uPA or activated TGF-beta. These data demonstrate that local tPA and uPA contribute to DTH regulation in allograft acceptor mice and suggest that these elements of the fibrinolytic pathway are used to control donor-reactive cell-mediated immunity in allograft acceptor mice. PMID- 10799872 TI - A new look at Syk in alpha beta and gamma delta T cell development using chimeric mice with a low competitive hematopoietic environment. AB - The Syk protein tyrosine kinase (PTK) is essential for B, but not T or NK, cell development, although certain T cell subsets (i.e., gamma delta T cells of intestine and skin) appear to be dependent on Syk. In this report, we have re evaluated the role of Syk in T cell development in hematopoietic chimeras generated by using Syk-deficient fetal liver hematopoietic stem cells (FL-HSC). We found that Syk-/- FL-HSC were vastly inferior to wild-type FL-HSC in reconstituting T cell development in recombinant-activating gene 2 (RAG2) deficient mice, identifying an unexpected and nonredundant role for Syk in this process. This novel function of Syk in T cell development was mapped to the CD44 CD25+ stage. According to previous reports, development of intestinal gamma delta T cells was arrested in Syk-/- -->RAG2-/- chimeras. In striking contrast, when hosts were the newly established alymphoid RAG2 x common cytokine receptor gamma chain (RAG2/gamma c) mice, Syk-/- chimeras developed intestinal gamma delta T cells as well as other T cell subsets (including alpha beta T cells, NK1.1+ alpha beta T cells, and splenic and thymic gamma delta T cells). However, all Syk deficient T cell subsets were reduced in number, reaching about 25-50% of controls. These results attest to the utility of chimeric mice generated in a low competitive hematopoietic environment to evaluate more accurately the impact of lethal mutations on lymphoid development. Furthermore, they suggest that Syk intervenes in early T cell development independently of ZAP-70, and demonstrate that Syk is not essential for the intestinal gamma delta T cell lineage to develop. PMID- 10799873 TI - Human vascular endothelial cells stimulate memory but not naive CD8+ T cells to differentiate into CTL retaining an early activation phenotype. AB - Endothelial cell (EC)-selective alloreactive CTL may mediate alloimmune vascular injury. In the present study, EC-selective CTL were generated in cocultures of purified human CD8+ T cells with allogeneic EC and were compared with conventional CTL against corresponding B lymphoblastoid cells (BLC). EC caused activation and expansion of memory but not naive CD8+ T cells, which differentiated into EC-selective CTL that retained high surface expression of CD69, CD25, and CD62L and displayed low intracellular perforin content. In contrast, BLC-stimulated CTL could be generated from naive or memory CD8+ T cells and showed a more mature phenotype (low CD69, CD25, and CD62L with higher levels of perforin). The expansion of alloreactive T cells by EC stimulation was 5- to 20-fold less effective than in corresponding BLC-stimulated cultures, accounting for a reduction in the assayable cytotoxicity of individual microcultures. In these IL-2-supplemented cocultures, no effect on CTL generation or phenotype was observed by mAb blocking of costimulation provided by LFA-3, ICAM-1, or CD40, by addition of comitogenic anti-CD28 mAb, or by preactivation of EC with CD40 ligand. Cyclosporine inhibited CTL expansion and cytotoxicity similarly in both EC- and BLC-stimulated cultures but did not affect the phenotype of those CTL that did emerge. This study extends the characterization of endothelium as an immunoregulatory cell type distinct from conventional APC and may explain why graft rejection within the arterial intima, an anatomic compartment in which EC may be the primary type of APC, is separable from rejection in the graft parenchyma. PMID- 10799874 TI - Differential requirement for p56lck in HIV-tat versus TNF-induced cellular responses: effects on NF-kappa B, activator protein-1, c-Jun N-terminal kinase, and apoptosis. AB - HIV-tat protein, like TNF, activates a wide variety of cellular responses, including NF-kappa B, AP-1, c-Jun N-terminal kinase (JNK), and apoptosis. Whether HIV-tat transduces these signals through the same mechanism as TNF is not known. In the present study we investigated the role of the T cell-specific tyrosine kinase p56lck in HIV-tat and TNF-mediated cellular responses by comparing the responses of Jurkat T cells with JCaM1 cells, an isogeneic lck-deficient T cell line. Treatment with HIV-tat protein activated NF-kappa B, degraded I kappa B alpha, and induced NF-kappa B-dependent reporter gene expression in a time dependent manner in Jurkat cells but not in JCaM1 cells, suggesting the critical role of p56lck kinase. These effects were specific to HIV-tat, as activation of NF-kappa B by PMA, LPS, H2O2, and TNF was minimally affected. p56lck was also found to be required for HIV-tat-induced but not TNF-induced AP-1 activation. Similarly, HIV-tat activated the protein kinases JNK and mitogen-activated protein kinase kinase in Jurkat cells but not in JCaM1 cells. HIV-tat also induced cytotoxicity, activated caspases, and reactive oxygen intermediates in Jurkat cells, but not in JCaM1 cells. HIV-tat activated p56lck activity in Jurkat cells. Moreover, the reconstitution of JCaM1 cells with p56lck tyrosine kinase reversed the HIV-tat-induced NF-kappa B activation and cytotoxicity. Overall, our results demonstrate that p56lck plays a critical role in the activation of NF kappa B, AP-1, JNK, and apoptosis by HIV-tat protein but has minimal or no role in activation of these responses by TNF. PMID- 10799875 TI - Monophosphoryl lipid A and QS21 increase CD8 T lymphocyte cytotoxicity to herpes simplex virus-2 infected cell proteins 4 and 27 through IFN-gamma and IL-12 production. AB - We have shown previously that IFN-gamma pretreatment of human epidermal cells (ECs) cultured in vitro partially reverses down-regulation of surface MHC class I by HSV infection, allowing recognition by CD8 CTLs, and that HSV immediate early (IE)/early (E) proteins are the predominant targets for CD8 CTLs. In this study of 25 subjects, CD8 CTLs recognized the HSV-2 IE infected cell protein 27 (ICP27) (expressed in autologous IFN-gamma-pretreated, Vaccinia virus recombinant infected ECs) in all subjects studied, ICP4 in 89%, and ICP0 in 11%. The main hierarchy of recognition was ICP27 > ICP4. ICP27 was the dominant target in 89% of subjects but showed great individual variability in the degree of cytotoxicity. CD8 cytotoxicity specific for HSV-2 IE proteins was enhanced by 48 67% when CD8 CTLs were coincubated with the combination of monophosphoryl lipid A and QS21 adjuvants at the time of Ag presentation. These adjuvants also significantly enhanced IL-12 and IFN-gamma production from nonadherent mononuclear cells stimulated by HSV-2-infected ECs. Addition of IL-12 and IFN gamma at the time of initial Ag presentation enhanced CD8 cytotoxicity to levels comparable with those stimulated by the adjuvants. Addition of neutralizing Abs to IL-12 or IFN-gamma inhibited CD8 T cell cytotoxicity up to 95% when a combination of the Abs were added at the time of initial Ag presentation. Therefore, the mechanism for the enhancement of CD8 T cell cytotoxicity by adjuvants in this system appears to be via increased levels of IL-12 and IFN gamma. PMID- 10799876 TI - Superoxide prevents nitric oxide-mediated suppression of helper T lymphocytes: decreased autoimmune encephalomyelitis in nicotinamide adenine dinucleotide phosphate oxidase knockout mice. AB - NO, which suppresses T cell proliferation, may be inactivated by superoxide (O2-) due to their strong mutual affinity. To examine this possibility, preactivated Th clones were cocultured with stimulated macrophages. PMA neutralized the inhibitory activity of NO, which was dependent on extracellular O2- production. In contrast, macrophages from p47phox -/- (pKO) mice, which lack functional NADPH oxidase, retained their NO-dependent inhibition of T cell proliferation upon stimulation with PMA, indicating that NADPH oxidase is the major source of NO inactivating O2- in this system. To examine the NO-O2- interaction in vivo, the role of NADPH oxidase in experimental autoimmune encephalomyelitis was studied in pKO mice. No clinical or histological signs were observed in the pKO mice. Neither a bias in Th subsets nor a reduced intensity of T cell responses could account for the disease resistance. Although spleen cells from pKO mice proliferated poorly in response to the immunogen, inhibition of NO synthase uncovered a normal proliferative response. These results indicate that NO activity may play a critical role in T cell responses in pKO mice and that in normal spleens inhibition of T cell proliferation by NO may be prevented by simultaneous NADPH oxidase activity. PMID- 10799877 TI - Alternate mucosal immune system: organized Peyer's patches are not required for IgA responses in the gastrointestinal tract. AB - The progeny of mice treated with lymphotoxin (LT)-beta receptor (LTbetaR) and Ig (LTbetaR-Ig) lack Peyer's patches but not mesenteric lymph nodes (MLN). In this study, we used this approach to determine the importance of Peyer's patches for induction of mucosal IgA Ab responses in the murine gastrointestinal tract. Immunohistochemical analysis revealed that LTbetaR-Ig-treated, Peyer's patch null (PP null) mice possessed significant numbers of IgA-positive (IgA+) plasma cells in the intestinal lamina propria. Further, oral immunization of PP null mice with OVA plus cholera toxin as mucosal adjuvant resulted in Ag-specific mucosal IgA and serum IgG Ab responses. OVA-specific CD4+ T cells of the Th2 type were induced in MLN and spleen of PP null mice. In contrast, when TNF and LT-alpha double knockout (TNF/LT-alpha-/-) mice, which lack both Peyer's patches and MLN, were orally immunized with OVA plus cholera toxin, neither mucosal IgA nor serum IgG anti-OVA Abs were induced. On the other hand, LTbetaR-Ig- and TNF receptor 55 Ig-treated normal adult mice elicited OVA- and cholera toxin B subunit-specific mucosal IgA responses, indicating that both LT-alphabeta and TNF/LT-alpha pathways do not contribute for class switching for IgA Ab responses. These results show that the MLN plays a more important role than had been appreciated until now for the induction of both mucosal and systemic Ab responses after oral immunization. Further, organized Peyer's patches are not a strict requirement for induction of mucosal IgA Ab responses in the gastrointestinal tract. PMID- 10799878 TI - An altered peptide ligand antagonizes antigen-specific T cells of patients with human T lymphotropic virus type I-associated neurological disease. AB - Human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is an inflammatory neurologic disease associated with HTLV I infection, in which chronically activated, HTLV-I-specific CD8+ CTL have been suggested to be immunopathogenic. In HLA-A2 HAM/TSP patients, CD8+ HTLV-I specific CTLs recognize an immunodominant peptide of the HTLV-I Tax protein, Tax11-19. We examined the functional outcome on activation of both cloned peripheral blood and cerebrospinal spinal fluid-derived CTL and bulk PBMC from HAM/TSP patients by altered peptide ligands (APL) derived from HTLV-I Tax11-19. In CTL clones generated from PBMC and CSF of HLA-A2 HAM/TSP patients, an APL substituted at position 5 significantly decreased CTL responses when compared with the native peptide. Moreover, these ligands were also shown to inhibit CTL responses to the native peptide in bulk PBMC of HLA-A2 HAM/TSP patients. These data suggest that a modification of an antigenic peptide at the central position can manipulate the T cell responses in bulk PBMC from different individuals with an inflammatory disease. Additionally, these results have implications for the potential use of APL-based immunotherapy in this T cell-mediated CNS disease. PMID- 10799879 TI - Cloning and characterization of human Lnk, an adaptor protein with pleckstrin homology and Src homology 2 domains that can inhibit T cell activation. AB - Lnk was originally cloned from a rat lymph node cDNA library and shown to participate in T cell signaling. Human Lnk (hLnk) was cloned by screening a Jurkat cell cDNA library. hLnk has a calculated molecular mass of 63 kDa, and its deduced amino acid sequence indicates the presence of an N-terminal proline-rich region, a pleckstrin homology domain, and a Src homology 2 domain. When expressed in COS cells, hLnk migrates with an apparent molecular mass of 75 kDa. Confocal fluorescence microscope analysis indicates that in COS cells transfected with an expression vector encoding a chimeric Lnk-green fluorescent protein, hLnk is found at the juxtanuclear compartment and also appears to be localized at the plasma membrane. Lnk is tyrosine-phosphorylated by p56lck. Following phosphorylation, p56lck binds to tyrosine-phosphorylated hLnk through its Src homology 2 domain. In COS cells cotransfected with hLnk, p56lck, and CD8-zeta, hLnk associated with tyrosine-phosphorylated TCR zeta-chain through its Src homology 2 domain. The overexpression of Lnk in Jurkat cells led to an inhibition of anti-CD3 mediated NF-AT-Luc activation. Our study reveals a potentially new mechanism of T cell-negative regulation. PMID- 10799880 TI - Inhibition of functional T cell priming and contact hypersensitivity responses by treatment with anti-secondary lymphoid chemokine antibody during hapten sensitization. AB - Recent studies have suggested a pivotal role for secondary lymphoid chemokine (SLC) in directing dendritic cell trafficking from peripheral to lymphoid tissues. As an extension of these studies, we examined the consequences of anti SLC Ab treatment during Ag priming on T cell function in an inflammatory response. We used a model of T cell-mediated inflammation, contact hypersensitivity (CHS), where priming of the effector T cells is dependent upon epidermal dendritic cell, Langerhans cells, and migration from the hapten sensitization site in the skin to draining lymph nodes. A single injection of anti-SLC Ab given at the time of sensitization with FITC inhibited Langerhans cell migration into draining lymph nodes for at least 3 days. The CHS response to hapten challenge was inhibited by anti-SLC Ab treatment in a dose-dependent manner. Despite the inhibition of CHS, T cells producing IFN-gamma following in vitro stimulation with anti-CD3 mAb or with hapten-labeled cells were present in the skin-draining lymph nodes of mice treated with anti-SLC Ab during hapten sensitization. These T cells were unable, however, to passively transfer CHS to naive recipients. Animals treated with anti-SLC Ab during hapten sensitization were not tolerant to subsequent sensitization and challenge with the hapten. In addition, anti-SLC Ab did not inhibit CHS responses when given at the time of hapten challenge. These results indicate an important role for SLC during sensitization for CHS and suggest a strategy to circumvent functional T cell priming for inflammatory responses through administration of an Ab inhibiting dendritic cell trafficking. PMID- 10799881 TI - Inducible expression of the gp49B inhibitory receptor on NK cells. AB - Murine NK cells express inhibitory receptors belonging to the C-type lectin-like (Ly-49, CD94/NKG2) and Ig superfamily-related (gp49) receptors. The murine gp49B receptor displays structural homology with human killer inhibitory receptors, and was previously identified to be a receptor on mast cells and activated NK cells. The gp49B receptor is highly related to gp49A, a receptor with unknown function. In this study, using a novel mAb produced against soluble gp49B molecules that cross-reacts with gp49A, we examined the cellular distribution and function of these receptors. gp49 is constitutively expressed on cells of the myeloid lineage throughout development, as well as on mature cells. Importantly, gp49 is not expressed on spleen- and liver-derived lymphocytes, including NK cells, but its expression is induced in vitro on NK cells following IL-2 stimulation, or in vivo by infection with murine CMV. Molecular studies revealed that both the immunoreceptor tyrosine-based inhibitory motif-containing gp49B as well as immunoreceptor tyrosine-based inhibitory motif-less gp49A receptors are up regulated on NK cells following murine CMV infection. When co-cross-linked with NK1.1, gp49B can inhibit NK1.1-mediated cytokine release by NK cells. Taken together, these studies demonstrate that the expression of gp49B on NK cells is regulated, providing the first example of an in vivo activation-induced NK cell inhibitory receptor, in contrast to the constitutively expressed Ly49 family. PMID- 10799882 TI - B lymphopoiesis in the thymus. AB - The thymus has been regarded as the major site of T cell differentiation. We find that in addition to alphabeta and gammadelta T cells, a significant number (approximately 3 x 104 per day) of B220+IgM+ mature B cells are exported from the thymus of C57BL/6 mice. Of these emigrating B cells, we estimate that at least approximately 2 x 104 per day are cells which developed intrathymically, whereas a maximum of approximately 0.8 x 104 per day are cells which circulated through the thymus from the periphery. The thymus possesses a significant number of pro-B and pre-B cells that express CD19, VpreB, lambda5, and pax-5. These B cell progenitors were found in the thymic cortex, whereas increasingly mature B cells were found in the corticomedullar and medullary regions. Other lymphoid cells, including NK cells and lymphoid dendritic cells, are not exported from the thymus at detectable levels. Thus, the thymus contributes to the formation of peripheral pools of B cells as well as of alphabeta and gammadelta T cells. PMID- 10799883 TI - Bcl-2 expression in synovial fibroblasts is essential for maintaining mitochondrial homeostasis and cell viability. AB - The regulation of proliferation and cell death is vital for homeostasis, but the mechanism that coordinately balances these events in rheumatoid arthritis (RA) remains largely unknown. In RA, the synovial lining thickens in part through increased proliferation and/or decreased synovial fibroblast cell death. Here we demonstrate that the anti-apoptotic protein, Bcl-2, is highly expressed in RA compared with osteoarthritis synovial tissues, particularly in the CD68-negative, fibroblast-like synoviocyte population. To determine the importance of endogenous Bcl-2, an adenoviral vector expressing a hammerhead ribozyme to Bcl-2 (Ad-Rbz-Bcl 2) mRNA was employed. Ad-Rbz-Bcl-2 infection resulted in reduced Bcl-2 expression and cell viability in synovial fibroblasts isolated from RA and osteoarthritis synovial tissues. In addition, Ad-Rbz-Bcl-2-induced mitochondrial permeability transition, cytochrome c release, activation of caspases 9 and 3, and DNA fragmentation. The general caspase inhibitor zVAD.fmk blocked caspase activation, poly(ADP-ribose) polymerase cleavage, and DNA fragmentation, but not loss of transmembrane potential or viability, indicating that cell death was independent of caspase activation. Ectopically expressed Bcl-xL inhibited Ad-Rbz-Bcl-2 induced mitochondrial permeability transition and apoptosis in Ad-Rbz-Bcl-2 transduced cells. Thus, forced down-regulation of Bcl-2 does not induce a compensatory mechanism to prevent loss of mitochondrial integrity and cell death in human fibroblasts. PMID- 10799885 TI - Nitric oxide synthase-2 and expression of perforin in uterine NK cells. AB - In human, mouse, and rat pregnancy, maternal NK cells accumulate and differentiate at implantation sites. These cells, termed uterine NK (uNK) cells, express NO synthase (NOS)-2 and develop cytolytic molecules such as perforin and granzymes during differentiation in situ. In this study, relationships between expression of the NOS-2 gene, uNK cell population density and tissue distribution, and synthesis of perforin were investigated. Uteri from wild-type (WT) and NOS-2-/- mice were collected at gestation days (g.d.) 8, 10, 12, 14, and 16 (n, >2/g.d.). Histochemical staining failed to reveal any differences between the population densities or tissue distributions of uNK cells in WT and NOS-2-/- uteri at any stage of gestation. By contrast, immunohistochemical staining with anti-perforin Abs demonstrated significantly fewer perforin-positive uNK cells in two uterine compartments of NOS-2-/- mice in comparison to the same compartments in WT mouse uteri. Perforin-positive uNK cells were reduced in NOS-2-/- metrial glands at g.d. 8, 10, and 12 and in decidua basalis at g.d. 12 (p < 0.05). Analysis of perforin protein by immunoblotting confirmed this observation. Northern blot hybridization studies showed that loss of perforin protein in NOS-2 /- mice was accompanied by decreased steady-state levels of perforin mRNA. These results demonstrate that migration of uNK cells into the uterus, selection of residency sites, and proliferation in situ are independent of NOS-2. By contrast, their differentiation, including transcription and translation of the cytotoxic molecule perforin, was shown to rely on normal expression of the NOS-2 gene. PMID- 10799884 TI - Absence of IFN-gamma or IL-12 has different effects on experimental myasthenia gravis in C57BL/6 mice. AB - Immunization with acetylcholine receptor (AChR) causes experimental myasthenia gravis (EMG). Th1 cells facilitate EMG development. IFN-gamma and IL-12 induce Th1 responses: we investigated whether these cytokines are necessary for EMG development. We immunized wild-type (WT) C57BL/6 mice and IFN-gamma and IL-12 knockout mutants (IFN-gamma-/-, IL-12-/-) with Torpedo AChR (TAChR). WT and IFN gamma-/- mice developed EMG with similar frequency, IL-12-/-mice were resistant to EMG. All strains synthesized anti-AChR Ab that were not IgM or IgE. WT mice had anti-AChR IgG1, IgG2b, and IgG2c, IFN-gamma-/- mice had significantly less IgG2c, and IL-12-/- mice less IgG2b and IgG2c. All mice had IgG bound to muscle synapses, but only WT and IFN-gamma-/- mice had complement; WT mice had both IgG2b and IgG2c, IFN-gamma-/- only IgG2b, and IL-12-/- neither IgG2b nor IgG2c. CD4+ cells from all AChR-immunized mice proliferated in response to AChR and recognized similar epitopes. After stimulation with TAChR, CD4+ cells from IFN gamma-/- mice secreted less IL-2 and similar amounts of IL-4 and IL-10 as WT mice. CD4+ cells from IL-12-/- mice secreted less IFN-gamma, but more IL-4 and IL 10 than WT mice, suggesting that they developed a stronger Th2 response to TAChR. The EMG resistance of IL-12-/- mice is likely due to both reduction of anti-TAChR Ab that bind complement and sensitization of modulatory Th2 cells. The reduced Th1 function of IFN-gamma-/- mice does not suffice to reduce all complement fixing IgG subclasses, perhaps because as in WT mice a protective Th2 response is missing. PMID- 10799886 TI - Retargeting T cell-mediated inflammation: a new perspective on autoantibody action. AB - To understand the pathogenesis of organ-specific autoimmune disease requires an appreciation of how the T cell-mediated inflammation is targeted, and how the organ function is compromised. In this study, autoantibody was documented to influence both of these parameters by modulating the distribution of T cell mediated inflammation. The murine autoimmune ovarian disease is induced by immunization with the ZP3330-342 peptide of the ovarian zona pellucida 3 glycoprotein, ZP3. Passively transferred or actively induced Ab to ZP3335-342 bound to the zona pellucida in the functional and degenerative ovarian follicles, and the ovaries remained histologically normal. Transfer of ZP3330-342 peptide specific T cells targeted the degenerative follicles and spared the functional follicles, and the resultant interstitial oophoritis was associated with unimpaired ovarian function. Unexpectedly, the coexistence of ZP3330-342 peptide specific T cells and zona-bound autoantibody led to a dramatic translocation of the ovarian inflammation to the growing and mature ovarian follicles, with destruction of the ovarian functional unit. Ab retargeted both Th1-induced mononuclear inflammation and Th2-induced eosinophilic inflammation, and retargeting was induced by murine and rat polyclonal Abs to multiple distinct native B cell determinants of the zona pellucida. Therefore, by reacting with the native determinants in tissue Ag, Ab alters the distribution of T cell-mediated inflammation, and results in destruction of the functional units of the target organ. We propose that this is a clinically important and previously unappreciated element of Ab action in autoimmune disease. PMID- 10799887 TI - Tolerance to cardiac allografts via local and systemic mechanisms after adenovirus-mediated CTLA4Ig expression. AB - Blockade of the CD28/B7 T cell costimulatory pathway prolongs allograft survival and induces tolerance in some animal models. We analyzed the efficacy of a CTLA4Ig-expressing adenovirus in preventing cardiac allorejection in rats, the mechanisms underlying heart transplant acceptance, and whether the effects of CTLA4Ig were restricted to the graft microenvironment or were systemic. CTLA4Ig gene transfer into the myocardium allowed indefinite graft survival (>100 days vs 9 +/- 1 days for controls) in 90% of cases, whereas CTLA4Ig protein injected systemically only prolonged cardiac allograft survival (by up to 22 days). CTLA4Ig could be detected in the graft and in the serum for at least 1 year after gene transfer. CTLA4Ig gene transfer induced local intragraft immunomodulation at day 5 after transplantation, as shown by decreased expression of the IL-2R and MHC II Ags; decreased levels of mRNA encoding for IFN-gamma, inducible NO synthase, and TGF-beta; and inhibited proliferative responses of graft infiltrating cells. Systemic immune responses were also down-modulated, as shown by the suppression of Ab production against donor alloantigens and cognate Ags, up to at least 120 days after gene transfer. Alloantigenic and mitogenic proliferative responses of graft-infiltrating cells and total splenocytes were inhibited and were not reversed by IL-2. In contrast, lymph node cells and T cells purified from splenocytes showed normal proliferation. Recipients of long term grafts treated with adenovirus coding for CTLA4Ig showed organ and donor specific tolerance. These data show that expression of CTLA4Ig was high and long lasting after adenovirus-mediated gene transfer. This expression resulted in down modulation of responses against cognate Ags, efficient suppression of local and systemic allograft immune responses, and ultimate induction of donor-specific tolerance. PMID- 10799888 TI - Transgenic human lambda 5 rescues the murine lambda 5 nullizygous phenotype. AB - The human lambda 5 (hu lambda 5) gene is the structural homologue of the murine lambda 5 (m lambda 5) gene and is transcriptionally active in pro-B and pre-B lymphocytes. The lambda 5 and VpreB polypeptides together with the Ig mu H chain and the signal-transducing subunits, Ig alpha and Ig beta, comprise the pre-B cell receptor. To further investigate the pro-B/pre-B-specific transcription regulation of hu lambda 5 in an in vivo model, we generated mouse lines that contain a 28-kb genomic fragment encompassing the entire hu lambda 5 gene. High levels of expression of the transgenic hu lambda 5 gene were detected in bone marrow pro-B and pre-B cells at the mRNA and protein levels, suggesting that the 28-kb transgene fragment contains all the transcriptional elements necessary for the stage-specific B progenitor expression of hu lambda 5. Flow cytometric and immunoprecipitation analyses of bone marrow cells and Abelson murine leukemia virus-transformed pre-B cell lines revealed the hu lambda 5 polypeptide on the cell surface and in association with mouse Ig mu and mouse VpreB. Finally, we found that the hu lambda 5 transgene is able to rescue the pre-B lymphocyte block when bred onto the m lambda 5-/- background. Therefore, we conclude that the hu lambda 5 polypeptide can biochemically and functionally substitute for m lambda 5 in vivo in pre-B lymphocyte differentiation and proliferation. These studies on the mouse and human pre-B cell receptor provide a model system to investigate some of the molecular requirements necessary for B cell development. PMID- 10799889 TI - IL-1 signaling cascade in liver cells and the involvement of a soluble form of the IL-1 receptor accessory protein. AB - The proinflammatory cytokine IL-1 induces the biosynthesis of a number of immunologically important proteins during infection, tissue damage, and/or stress, in part through the activation of the transcription factor NF-kappaB. Signal transduction is initiated at the cell membrane by complex formation between extracellular IL-1 and the transmembrane IL-1R type I (IL-1RI) and IL-1R accessory protein (IL-1RAcP). The intracellular signaling cascade involves recruitment of two IL-1R-associated kinases, IRAK1 and IRAK2, and the adapter protein MyD88, events which are dependent on the intracellular domain of membrane bound IL-1RAcP (mIL-1RAcP). In mouse liver, IL-1RAcP is expressed as a soluble protein (sIL-1RAcP), the function of which is unknown. We have cloned the human sIL-1RAcP and established by sequence analysis that the human sIL-1RAcP mRNA arises from alternative splicing of the IL-1RAcP gene (shown here to encompass 12 exons spanning more than 56 kb). Furthermore, we demonstrate that human HepG2 hepatoma cells express both mIL-1RAcP and sIL-1RAcP and that signal transduction in these cells is mediated through IRAK1, IRAK2, and MyD88. We show that phorbol esters induce a change in the pre-mRNA splice pattern such that sIL-1RAcP mRNA becomes the dominant form. Overexpression of a membrane-anchored fusion protein of sIL-1RAcP and MHC in HepG2 cells inhibits IL-1-mediated NF-kappaB activation, whereas coexpression of IL-1RI with membrane-anchored sIL-1RAcP restores the capacity of the cells to respond to IL-1. This suggests that sIL-1RAcP may act as an inhibitor of IL-1 by directly interacting with IL-1RI to abolish its capacity to transduce signal. PMID- 10799890 TI - Regulation of alternative splicing of CD45 by antagonistic effects of SR protein splicing factors. AB - CD45 is a transmembrane glycoprotein possessing tyrosine phosphatase activity, which is involved in cell signaling. CD45 is expressed on the surface of most leukocytes and can be alternatively spliced by the inclusion or skipping of three variable exons (4, 5, and 6 or A, B, and C) to produce up to eight isoforms. In T cells, the splicing pattern of CD45 isoforms changes after activation; naive cells express high m.w. isoforms of CD45 which predominantly express exon A (CD45RA), whereas activated cells lose expression of exon A to form low m.w. isoforms of CD45 including CD45RO. Little is known about the specific factors controlling the switch in CD45 splicing which occurs on activation. In this study, we examined the influence of the SR family of splicing factors, which, like CD45, are expressed in tissue-specific patterns and have been shown to modulate the alternative splicing of a variety of transcripts. We show that specific SR proteins have antagonistic effects on CD45 splicing, leading either to exon inclusion or skipping. Furthermore, we were able to demonstrate specific changes in the SR protein expression pattern during T cell activation. PMID- 10799891 TI - A contig map of the Mhc class I genomic region in the zebrafish reveals ancient synteny. AB - In contrast to the human and mouse Mhc, in which the clusters of class I and class II loci reside in close vicinity to one another, in the zebrafish, Danio rerio, they are found in different linkage groups. Chromosome walking using BAC (bacterial artificial chromosome) and PAC (P1 artificial chromosome) clones reveals the zebrafish class I region to occupy a segment of approximately 450 kb and to encompass at least 19 loci. These include three class I (Dare-UDA, -UEA, UFA), five proteasome subunit beta (PSMB8, -9A, -9C, -11, -12), two TAPs (TAP2A, TAP2B), and one TAP binding protein (TAPBP). This arrangement contrasts with the arrangements found in human and mouse Mhc, in which the orthologues of the PSMB, TAP, and TAPBP loci reside within the class II region. In addition to this main zebrafish class I contig, a shorter contig of about 150 kb contains two additional class I (UBA, UCA) and at least five other loci. It probably represents a different haplotype of part of the class I region. The previously identified UAA gene shares an identical 5' part with UEA, but the two genes differ in their 3' parts. One of them is probably the result of an unequal crossing over. The described organization has implications for the persistence of syntenic relationships, coevolution of loci, and interpretation of the origin of the human/mouse Mhc organization. PMID- 10799892 TI - Role of OCA-B in 3'-IgH enhancer function. AB - OCA-B (alternately called Bob1 and OBF-1) is a B cell-specific coactivator that interacts with the ubiquitously expressed Oct-1 and the B cell-restricted Oct-2 to activate transcription via the octamer site (5'-ATGCAAAT-3'). OCA-B-/- mice appear to undergo normal Ag-independent B cell maturation. However, Ag-dependent B cell differentiation, including germinal center formation, production of secondary Ig isotypes, and proliferation in response to surface Ig cross-linking, is greatly affected. We demonstrate that the observed reductions in expression of class-switched isotypes in OCA-B-/- mice may be due in part to deficiencies in the function of the 3'-IgH enhancer elements. Furthermore, we find that surface Ig cross-linking represses all the Ig enhancers and that this repression is absent in OCA-B-/- B cells. These results suggest an important role for OCA-B in Ig enhancer function in vivo. PMID- 10799893 TI - The IgG Fc contains distinct Fc receptor (FcR) binding sites: the leukocyte receptors Fc gamma RI and Fc gamma RIIa bind to a region in the Fc distinct from that recognized by neonatal FcR and protein A. AB - The CH2-CH3 interface of the IgG Fc domain contains the binding sites for a number of Fc receptors including Staphylococcal protein A and the neonatal Fc receptor (FcRn). It has recently been proposed that the CH2-CH3 interface also contains the principal binding site for an isoform of the low affinity IgG Fc receptor II (Fc gamma RIIb). The Fc gamma RI and Fc gamma RII binding sites have previously been mapped to the lower hinge and the adjacent surface of the CH2 domain although contributions of the CH2-CH3 interface to binding have been suggested. This study addresses the question whether the CH2-CH3 interface plays a role in the interaction of IgG with Fc gamma RI and Fc gamma RIIa. We demonstrate that recombinant soluble murine Fc gamma RI and human Fc gamma RIIa did not compete with protein A and FcRn for binding to IgG, and that the CH2-CH3 interface therefore appears not to be involved in Fc gamma RI and Fc gamma RIIa binding. The importance of the lower hinge was confirmed by introducing mutations in the proposed binding site (LL234,235AA) which abrogated binding of recombinant soluble Fc gamma RIIa to human IgG1. We conclude that the lower hinge and the adjacent region of the CH2 domain of IgG Fc is critical for the interaction between Fc gamma RIIa and human IgG, whereas contributions of the CH2-CH3 interface appear to be insignificant. PMID- 10799894 TI - Structural analysis of CTLA-4 function in vivo. AB - CTLA-4-mediated inhibition of T cell activation may be accomplished by competition for ligands and/or by signals mediated through the intracellular domain. Studies have implicated Tyr201 in the cytoplasmic domain of CTLA-4 in regulating CTLA-4 signal transduction and intracellular trafficking. To investigate the mechanism of CTLA-4 function in vivo, transgenes encoding wild type CTLA-4 (FL), a mutant lacking the cytoplasmic domain of CTLA-4 (DeltaCTLA-4 tail), or a CTLA-4 Tyr201 mutant (Y201V) were introduced into CTLA-4-deficient mice. CTLA-4-/- mice display an autoimmune lymphoproliferative disorder resulting in tissue destruction and early death. When either the FL or the Y201V transgene was bred into CTLA-4-/- animals, a complete rescue from lymphoproliferation and autoimmunity was observed. In contrast, CTLA-4-/- mice expressing the DeltaCTLA-4 tail transgene were long lived with no evidence of multiorgan lymphocytic infiltration, but exhibited lymphadenopathy and accumulated large numbers of activated T cells. Furthermore, these animals displayed a Th2-biased phenotype which conferred susceptibility to Leishmania infection. These results indicate that the inhibitory effect of CTLA-4 is mediated in part through the ability of the extracellular domain to compete for ligands. The cytoplasmic domain of CTLA 4, however, is required for complete inhibitory function of the receptor and for regulation of Th cell differentiation in vivo. PMID- 10799895 TI - Human C4b-binding protein has overlapping, but not identical, binding sites for C4b and streptococcal M proteins. AB - Many strains of Streptococcus pyogenes bind C4b-binding protein (C4BP), an inhibitor of complement activation. The binding is mediated by surface M proteins in a fashion that has been suggested to mimic the binding of C4b. We have previously shown that a positively charged cluster at the interface between complement control protein domains 1 and 2 of C4BP alpha-chain is crucial for the C4b-C4BP interaction. To extend this observation, and to investigate the interaction with M proteins, we constructed and characterized a total of nine mutants of C4BP. We identified a key recognition surface for M proteins that overlaps with the C4b binding site because substitution of R64 and H67 by Gln dramatically reduces binding to both ligands. However, the analysis of all mutants indicates that the binding sites for C4b and M proteins are only overlapping, but not identical. Furthermore, M proteins were able to displace C4BP from immobilized C4b, whereas C4b only weakly affected binding of C4BP to immobilized M proteins. We found that the molecular mechanisms involved in these two interactions differ because the binding between M proteins and C4BP is relatively insensitive to salt in contrast to the C4BP-C4b binding. In addition, six mAbs directed against the alpha-chain interfered with C4b-C4BP interaction, whereas only two of them efficiently inhibited binding of C4BP to M proteins. Collectively, our results suggest that binding between C4b and C4BP is governed mostly by electrostatic interactions, while additional noncovalent forces cause tight binding of C4BP to streptococcal M proteins. PMID- 10799896 TI - Repeated exposure induces periportal fibrosis in Schistosoma mansoni-infected baboons: role of TGF-beta and IL-4. AB - Recently, we observed that repeated Schistosoma mansoni infection and treatment boost Th2-associated cytokines and TGF-beta production in baboons. Other studies have shown that some chronically infected baboons develop hepatic fibrosis. Because TGF-beta, IL-2, and IL-4 have been shown to participate in development of fibrosis in murine schistosomiasis, the present study examined whether repeated exposure stimulates hepatic fibrosis in olive baboons. To test this hypothesis, animals were exposed to similar numbers of S. mansoni cercariae given once or repeatedly. After 19 wk of infection, animals were cured with praziquantel and reinfected once or multiple times. Hepatic granulomatous inflammation and fibrosis were assessed from serial liver biopsies taken at weeks 6, 9, and 16 after reinfection and egg Ag (schistosome egg Ag)-specific cytokine production by PBMC were measured simultaneously. Periportal fibroblast infiltration and extracellular matrix deposition (fibrosis), angiogenesis, and biliary duct hyperplasia developed in some animals. The presence and amount of fibrosis directly correlated with the frequency of exposure. Fibrosis was not associated with adult worm or tissue egg burden. The amount of fibrosis correlated with increased schistosome egg Ag-driven TGF-beta at 6, 9, and 16 wk postinfection (rs = 0.9, 0.8, and 0.54, respectively, all p < 0.01) and IL-4 production (p = 0.02) at 16 wk postinfection and not IFN-gamma, IL-2, IL-5, or IL-10. These data suggest that repeated exposure is a risk factor for periportal fibrosis by a mechanism that primes lymphocytes to produce increased levels of profibrotic molecules that include TGF-beta and IL-4. PMID- 10799897 TI - Arthropod- and host-specific Borrelia burgdorferi bbk32 expression and the inhibition of spirochete transmission. AB - Antisera to BBK32 (a Borrelia burgdorferi fibronectin-binding protein) and BBK50, two Ags synthesized during infection, protect mice from experimental syringe borne Lyme borreliosis. Therefore, B. burgdorferi bbk32 and bbk50 expression within Ixodes scapularis ticks and the murine host, and the effect of BBK32 and BBK50 antisera on spirochetes throughout the vector-host life cycle were investigated. bbk32 and bbk50 mRNA and protein were first detected within engorged ticks, demonstrating regulated expression within the vector. Then bbk32 expression increased in mice at the cutaneous site of inoculation. During disseminated murine infection, bbk32 and bbk50 were expressed in several murine tissues, and mRNA levels were greatest in the heart and spleen at 30 days. BBK32 antisera protected mice from tick-borne B. burgdorferi infection and spirochete numbers were reduced by 90% within nymphs that engorged on immunized mice. Moreover, 75% of these ticks did not retain spirochetes upon molting, and subsequent B. burgdorferi transmission by adult ticks was impaired. Larval acquisition of B. burgdorferi by I. scapularis was also inhibited by BBK32 antisera. These data demonstrate that bbk32 and bbk50 are expressed during tick engorgement and that BBK32 antisera can interfere with spirochete transmission at various stages of the vector-host life cycle. These studies provide insight into mechanisms of immunity to Lyme borreliosis and other vector-borne diseases. PMID- 10799898 TI - Regulation of RANTES chemokine gene expression requires cooperativity between NF kappa B and IFN-regulatory factor transcription factors. AB - Virus infection of host cells activates a set of cellular genes, including cytokines, IFNs, and chemokines, involved in antiviral defense and immune activation. Previous studies demonstrated that virus-induced transcriptional activation of a member of the human CC-chemokine RANTES required activation of the latent transcription factors IFN-regulatory factor (IRF)-3 and NF-kappa B via posttranslational phosphorylation. In the present study, we further characterized the regulatory control of RANTES transcription during virus infection using in vivo genomic footprinting analyses. IRF-3, the related IRF-7, and NF-kappa B are identified as important in vivo binding factors required for the cooperative induction of RANTES transcription after virus infection. Using fibroblastic or myeloid cells, we demonstrate that the kinetics and strength of RANTES virus induced transcription are highly dependent on the preexistence of IRFs and NF kappa B. Use of dominant negative mutants of either I kappa B-alpha or IRF-3 demonstrate that disruption of either pathway dramatically abolishes the ability of the other to bind and activate RANTES expression. Furthermore, coexpression of IRF-3, IRF-7, and p65/p50 leads to synergistic activation of RANTES promoter transcription. These studies reveal a model of virus-mediated RANTES promoter activation that involves cooperative synergism between IRF-3/IRF-7 and NF-kappa B factors. PMID- 10799899 TI - Pivotal role of the CC chemokine, macrophage-derived chemokine, in the innate immune response. AB - Macrophage-derived chemokine (MDC), a recently identified CC chemokine, has been regarded to be involved in chronic inflammation and dendritic cell and lymphocyte homing. In this study, we demonstrate a pivotal role for MDC during experimental sepsis induced by cecal ligation and puncture (CLP). Intraperitoneal administration of MDC (1 microg/mouse) protected mice from CLP-induced lethality. The survival was accompanied by increased number of peritoneal macrophages and decreased recovery of viable bacteria from the peritoneum and peripheral blood. In addition, mice treated with an i.p. injection of MDC cleared bacteria more effectively than those in the control when 3 x 108 CFU live Escherichia coli was i.p. inoculated. Endogenous MDC was detected in the peritoneum after CLP, and neutralization of the MDC with anti-MDC Abs decreased CLP-induced recruitment of peritoneal macrophages and increased the recovery of viable bacteria from the peritoneum and peripheral blood. MDC blockade was deleterious in the survival of mice after CLP. In vitro, MDC enhanced the phagocytic and killing activities of peritoneal macrophages to E. coli and induced both a respiratory burst and the release of lysozomal enzyme from macrophages. Furthermore, MDC dramatically ameliorated CLP-induced systemic tissue inflammation as well as tissue dysfunction, which were associated in part with decreased levels of TNF-alpha, macrophage inflammatory proteins-1alpha and -2, and KC in specific tissues. Collectively, these results indicate novel regulatory activities of MDC in innate immunity during sepsis and suggest that MDC may aid in an adjunct therapy in sepsis. PMID- 10799900 TI - Modulation of HIV-1 replication by a novel RhoA effector activity. AB - The RhoA GTPase is involved in regulating actin cytoskeletal organization, gene expression, cell proliferation, and survival. We report here that p115-RhoGEF, a specific guanine nucleotide exchange factor (GEF) and activator of RhoA, modulates HIV-1 replication. Ectopic expression of p115-RhoGEF or Galpha13, which activates p115-RhoGEF activity, leads to inhibition of HIV-1 replication. RhoA activation is required and the inhibition affects HIV-1 gene expression. The RhoA effector activity in inhibiting HIV-1 replication is genetically separable from its activities in transformation of NIH3T3 cells, activation of serum response factor, and actin stress fiber formation. These findings reveal that the RhoA signal transduction pathway regulates HIV-1 replication and suggest that RhoA inhibits HIV-1 replication via a novel effector activity. PMID- 10799901 TI - IL-10 is required for prevention of necrosis in the small intestine and mortality in both genetically resistant BALB/c and susceptible C57BL/6 mice following peroral infection with Toxoplasma gondii. AB - The role for IL-10 in the immunopathogenesis of acute toxoplasmosis following peroral infection was examined in both genetically susceptible C57BL/6 and resistant BALB/c mice. C57BL/6-background IL-10-targeted mutant (IL-10-/-) mice all died in 2 wk after infection with 20 cysts of the ME49 strain, whereas only 20% of control mice succumbed. Histological studies revealed necrosis in the small and large intestines and livers of infected IL-10-/- mice. The necrosis in the small intestine was the most severe pathologic response and was not observed in control mice. Treatment of infected IL-10-/- mice with either anti-CD4 or anti IFN-gamma mAb prevented intestinal pathology and significantly prolonged time to death. Treatment of these animals with anti-IL-12 mAb also prevented the pathology. Significantly greater amounts of IFN-gamma mRNA were detected in the lamina propria lymphocytes obtained from the small intestine of infected IL-10-/- mice than those from infected control mice. In common with C57BL/6-background IL 10-/- mice, BALB/c-background IL-10-/- mice all died developing intestinal pathology after infection. Control BALB/c mice all survived even after infection with 100 cysts and did not develop the intestinal lesions. Treatment with anti IFN-gamma mAb prevented the pathology and prolonged time to death of the infected IL-10-/- mice. These results strongly suggest that IL-10 plays a critical role in down-regulating IFN-gamma production in the small intestine following sublethal peroral infection with Toxoplasma gondii and that this down-regulatory effect of IL-10 is required for prevention of development of IFN-gamma-mediated intestinal pathology and mortality in both genetically resistant BALB/c and susceptible C57BL/6 mice. PMID- 10799902 TI - Expression of IFN-inducible T cell alpha chemoattractant by human endothelial cells is cyclosporin A-resistant and promotes T cell adhesion: implications for cyclosporin A-resistant immune inflammation. AB - IFN-inducible T cell alpha chemoattractant (I-TAC) is a recently discovered member of the CXC chemokine family. It is a potent T cell chemoattractant expressed by IFN-gamma-treated astrocytes, monocytes, keratinocytes, bronchial epithelial cells, and neutrophils. In this study, we show that I-TAC is also expressed by IFN-gamma-treated endothelial cells (EC), both at the mRNA and protein levels. Induction of the I-TAC message is rapid and sustained over 24 h. TNF-alpha does not induce I-TAC mRNA alone, but does act synergistically with IFN gamma. Blocking Abs to I-TAC, or to its receptor, CXCR3, reduce T cell adhesion to EC monolayers demonstrating that the expressed protein is functional. Finally, the expression of I-TAC by EC is resistant to the immunosuppressive drug cyclosporin A, suggesting that I-TAC may contribute to the chronic immune inflammation characteristic of graft arteriosclerosis. PMID- 10799904 TI - Cellular regulation of cytosolic group IV phospholipase A2 by phosphatidylinositol bisphosphate levels. AB - Cytosolic group IV phospholipase A2 (cPLA2) is a ubiquitously expressed enzyme with key roles in intracellular signaling. The current paradigm for activation of cPLA2 by stimuli proposes that both an increase in intracellular calcium and mitogen-activated protein kinase-mediated phosphorylation occur together to fully activate the enzyme. Calcium is currently thought to be needed for translocation of the cPLA2 to the membrane via a C2 domain, whereas the role of cPLA2 phosphorylation is less clearly defined. Herein, we report that brief exposure of P388D1 macrophages to UV radiation results in a rapid, cPLA2-mediated arachidonic acid mobilization, without increases in intracellular calcium. Thus, increased Ca2+ availability is a dispensable signal for cPLA2 activation, which suggests the existence of alternative mechanisms for the enzyme to efficiently interact with membranes. Our previous in vitro data suggested the importance of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) in the association of cPLA2 to model membranes and hence in the regulation of cPLA2 activity. Experiments described herein show that PtdInsP2 also serves a similar role in vivo. Moreover, inhibition of PtdInsP2 formation during activation conditions leads to inhibition of the cPLA2-mediated arachidonic acid mobilization. These results suggest that cellular PtdInsP2 levels are involved in the regulation of group IV cPLA2 activation. PMID- 10799903 TI - Glycosphingolipid-induced relocation of Lyn and Syk into detergent-resistant membranes results in mast cell activation. AB - Sphingosine, sphingosine-1-phosphate, and the more complex sphingolipid ceramide exert strong immunomodulatory effects on a variety of leukocytes. However, little is known regarding such a potential of glycosphingolipids, a class of sugar derivatives of sphingosine. Here we demonstrate that galactosylsphingosine, one of the smallest representatives of this group, accumulates in the detergent resistant membranes resulting in the relocation of the tyrosine kinases Lyn and Syk into this compartment. The result of this is an enhanced tyrosine phosphorylation and kinase activity leading to priming and activation of mast cells by conveying a weak yet significant activation of the mitogen-activated protein kinase pathway(s). In comparison to IgE/Ag triggering, galactosylsphingosine stimulates the mitogen-activated protein kinase pathway more rapidly and favors c-Jun NH2-terminal kinase 1 activation over extracellular signal-regulatory kinase 1 and 2. At the transcription factor level, this "ultratransient signaling event" results in an activation of JunD as the predominant AP-1 component. In this respect, the effects of galactosylsphingosine are clearly distinct from the signaling elicited by other sphingolipids without the sugar moiety, such as sphingosine-1-phosphate. PMID- 10799905 TI - Prostaglandin E2 stimulates AP-1-mediated CD14 expression in mouse macrophages via cyclic AMP-dependent protein kinase A. AB - PGs play a functional role in the early stage of Gram-negative bacterial infections, because this prostanoid is produced rapidly by epithelial cells after a bacterial infection. CD14, one of the LPS receptors, is a key molecule in triggering the response to bacterial LPS in association with a Toll-like molecule. Therefore, in this study, we investigated the effect of PG on CD14 expression in mouse macrophages. PGE1, PGE2, and PGA1 among the PGs tested strongly stimulated the expression of the CD14 gene in the cells. The stimulatory action also was observed by Western blot analysis. cAMP-elevating agents stimulated expression of CD14 gene as well. Protein kinase A inhibitor, N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), but not protein kinase C inhibitor 3-(1-[3-(dimethylamino)propyl]-1H-indol-3-yl)-4-(1H-indol-3 yl)-1H-py rrole-2,5-dione (GF109203X), abolished the stimulated expression of CD14. A run-on assay showed that PGE2 stimulated the CD14 gene expression at the transcriptional level via protein kinase A. PGE2 also stimulated activation of AP 1, a heterodimer of c-Jun and c-Fos, because the prostanoid increased specific binding of nuclear proteins to the AP-1 consensus sequence and stimulated AP-1 promoted luciferase activity. PGE2-stimulated expression of CD14 was inhibited by antisense c-fos and c-jun oligonucleotides, but not by their sense oligonucleotides. Finally, PGE2 pretreatment synergistically stimulated LPS induced expression of IL-1beta and IL-6 genes in mouse macrophages. Therefore, the present study demonstrates that PGE2 has the ability to stimulate AP-1 mediated expression of CD14 in mouse macrophages via cAMP-dependent protein kinase A. PMID- 10799906 TI - Inhibition of antigen-induced eosinophilia and late phase airway hyperresponsiveness by an IL-5 antisense oligonucleotide in mouse models of asthma. AB - Chronic airway eosinophilia is associated with allergic asthma and is mediated in part by secretion of IL-5 from allergen-specific Th2 lymphocytes. IL-5 is a known maturation and antiapoptotic factor for eosinophils and stimulates release of nascent eosinophils from bone marrow into the peripheral circulation. An antisense oligonucleotide found to specifically inhibit IL-5 expression in vitro was observed to significantly reduce experimentally induced eosinophilia in vivo, in both the murine OVA lung challenge and allergic peritonitis models. Intravenous administration resulted in sequence-dependent inhibition of eosinophilia coincident with reduction of IL-5 protein levels, supporting an antisense mechanism of action. Potent suppression of lung eosinophilia was observed up to 17 days after cessation of oligonucleotide dosing, indicating achievement of prolonged protection with this strategy. Furthermore, sequence specific, antisense oligonucleotide-mediated inhibition of Ag-mediated late phase airway hyperresponsiveness was also observed. These data underscore the potential utility of an antisense approach targeting IL-5 for the treatment of asthma and eosinophilic diseases. PMID- 10799907 TI - Anti-inflammatory effect of heat shock protein induction is related to stabilization of I kappa B alpha through preventing I kappa B kinase activation in respiratory epithelial cells. AB - Heat shock protein (HSP) induction confers protection against diverse forms of cellular and tissue injury. However, the mechanism by which HSP exerts cytoprotective effects is unclear. Because HSP induction inhibits genetic expression of pro-inflammatory cytokines, the transcription of which is dependent on NF-kappa B activation, we explored the relationship between the anti inflammatory effect of HSP induction and the NF-kappa B/I kappa B alpha pathway. Both HS and sodium arsenite treatment increased HSP70 expression time dependently at mRNA and protein levels. Prior induction of HSP suppressed cytokine-induced IL 8 and TNF-alpha expression at both mRNA and protein levels. Although HSP induction did not affect total cellular expression of NF-kappa B, TNF-alpha induced increase in NF-kappa B-DNA binding activity and nuclear translocation of the p65 subunit of NF-kappa B were inhibited by prior HSP induction, suggesting that activation of NF-kappa B was blocked. Cytokine-induced I kappa B alpha phosphorylation and its degradation were blocked in HSP-induced cells. Immune complex kinase assays demonstrated that TNF-alpha induced increase in I kappa B kinase activity was suppressed by prior HSP induction. These results suggest that the anti-inflammatory effect of HSP induction in respiratory epithelial cells is related to stabilization of I kappa B alpha, possibly through the prevention of I kappa B kinase activation, which thereby inhibits activation of NF-kappa B. PMID- 10799908 TI - Neurokinin-1 receptor agonists are involved in mediating neutrophil accumulation in the inflamed, but not normal, cutaneous microvasculature: an in vivo study using neurokinin-1 receptor knockout mice. AB - We have used tachykinin neurokinin-1 receptor (NK1 receptor) knockout mice to learn of the link between NK1 receptors and neutrophil accumulation in normal naive skin, as compared with inflamed skin. Intradermal substance P (300 pmol) induced edema formation in wild-type mice, but not in NK1 knockout mice, as expected. However, in contrast to IL-1beta (0.3 pmol), substance P did not induce neutrophil accumulation in wild-type mice. IL-1beta-induced neutrophil accumulation was similar in wild-type and knockout mice, but a significant (p < 0.05) contributory effect of added NK1 agonists, which by themselves have no effect on neutrophil accumulation in normal skin, was observed. The results support the concept that NK1 agonists such as substance P cannot act on their own to mediate neutrophil accumulation in naive skin and provide direct evidence that in inflamed skin, under certain circumstances, the NK1 receptor can play a pivotal role in modulating neutrophil accumulation during the ongoing inflammatory process. We investigated responses to two inflammatory stimuli (carrageenin and zymosan). Neutrophil accumulation was significantly attenuated (p < 0.001) in carrageenin- but not zymosan-induced inflammation in NK1 knockout mice. The carrageenin (500 microg)-induced response was inhibited (p < 0.05) by a NK1 receptor antagonist, SR140333 (480 nmol/kg i.v. at -5 min), in the wild-type group. The bradykinin B1 and B2 receptor antagonists (desArg9[Leu8]bradykinin and HOE 140) each reduced neutrophil accumulation to carrageenin in wild-type animals (p < 0.05), but did not cause further reduction of the suppressed response of knockout mice. The results provide evidence that kinin receptors participate in NK1 receptor-dependent neutrophil accumulation in inflamed mouse skin. PMID- 10799909 TI - Na+-dependent high-affinity glutamate transport in macrophages. AB - Excessive accumulation of glutamate in the CNS leads to excitotoxic neuronal damage. However, glutamate clearance is essentially mediated by astrocytes through Na+-dependent high-affinity glutamate transporters (excitatory amino acid transporters (EAATs)). Nevertheless, EAAT function was recently shown to be developmentally restricted in astrocytes and undetectable in mature astrocytes. This suggests a need for other cell types for clearing glutamate in the brain. As blood monocytes infiltrate the CNS in traumatic or inflammatory conditions, we addressed the question of whether macrophages expressed EAATs and were involved in glutamate clearance. We found that macrophages derived from human blood monocytes express both the cystine/glutamate antiporter and EAATs. Kinetic parameters were similar to those determined for neonatal astrocytes and embryonic neurons. Freshly sorted tissue macrophages did not possess EAATs, whereas cultured human spleen macrophages and cultured neonatal murine microglia did. Moreover, blood monocytes did not transport glutamate, but their stimulation with TNF-alpha led to functional transport. This suggests that the acquisition of these transporters by macrophages could be under the control of inflammatory molecules. Also, monocyte-derived macrophages overcame glutamate toxicity in neuron cultures by clearing this molecule. This suggests that brain-infiltrated macrophages and resident microglia may acquire EAATs and, along with astrocytes, regulate extracellular glutamate concentration. Moreover, we showed that EAATs are involved in the regulation of glutathione synthesis by providing intracellular glutamate. These observations thus offer new insight into the role of macrophages in excitotoxicity and in their response to oxidative stress. PMID- 10799910 TI - Effect of CD14 blockade in rabbits with Escherichia coli pneumonia and sepsis. AB - CD14, a pattern recognition receptor found on myeloid cells, is a critical component of the innate immune system that mediates local and systemic host responses to Gram-negative and Gram-positive bacterial products. Previous studies in normal animals have tested the effect of CD14 blockade on the systemic response to i.v. LPS. The goals of the study were to determine whether CD14 blockade protected against the deleterious systemic response associated with Escherichia coli pneumonia and to determine whether this strategy affected the pulmonary response to tissue infection. Rabbits were pretreated with either anti CD14 mAb or isotype control mAb at 2.5 mg/kg. E. coli (1 x 109 CFU) was inoculated into the lungs, and the animals were observed for either 4 or 24 h. The blockade of CD14 improved the mean arterial blood pressure (p = 0.001) and decreased the i.v. fluid requirements (p = 0.01). Although this therapy protected the vascular compartment, rabbits treated with anti-CD14 mAb had increased bacterial burdens in the bronchoalveolar lavage fluid recovered from the instilled lung (p = 0.005) and widened alveolar-arterial oxygen difference. Blockade of CD14 prevents the deleterious systemic responses that occur in sepsis; however, other measures are necessary to control bacterial proliferation at the primary site of infection. PMID- 10799911 TI - Complement C1q is dramatically up-regulated in brain microglia in response to transient global cerebral ischemia. AB - Recent evidence suggests that the pathophysiology of neurodegenerative and inflammatory neurological diseases has a neuroimmunological component involving complement, an innate humoral immune defense system. The present study demonstrates the effects of experimentally induced global ischemia on the biosynthesis of C1q, the recognition subcomponent of the classical complement activation pathway, in the CNS. Using semiquantitative in situ hybridization, immunohistochemistry, and confocal laser scanning microscopy, a dramatic and widespread increase of C1q biosynthesis in rat brain microglia (but not in astrocytes or neurons) within 24 h after the ischemic insult was observed. A marked increase of C1q functional activity in cerebrospinal fluid taken 1, 24, and 72 h after the ischemic insult was determined by C1q-dependent hemolytic assay. In the light of the well-established role of complement and complement activation products in the initiation and maintenance of inflammation, the ischemia-induced increase of cerebral C1q biosynthesis and of C1q functional activity in the cerebrospinal fluid implies that the proinflammatory activities of locally produced complement are likely to contribute to the pathophysiology of cerebral ischemia. Pharmacological modulation of complement activation in the brain may be a therapeutic target in the treatment of stroke. PMID- 10799912 TI - Induction of functional anaphylatoxin C5a receptors on hepatocytes by in vivo treatment of rats with IL-6. AB - In normal rat liver, anaphylatoxin C5a receptors (C5aR) are only expressed by nonparenchymal cells, mainly Kupffer cells and hepatic stellate cells, but not by parenchymal cells, i.e., hepatocytes (HC). Nevertheless, C5a stimulates glucose output by HC. This HC-specific defense reaction is induced indirectly via prostanoids secreted by the C5aR-expressing Kupffer cells and hepatic stellate cells. It is shown here that under inflammatory conditions simulated by in vivo treatment of rats with IL-6 C5aR mRNA and protein were induced in HC in a time dependent manner. Maximal mRNA and protein expression were observed at 4-8 h and 8-10 h, respectively, after IL-6 injection. The newly expressed receptors were functional, because recombinant rat C5a significantly activated glycogen phosphorylase in HC isolated from IL-6-treated but not in HC from control rats. In perfused livers of IL-6-treated animals in contrast to control animals, recombinant rat C5a-induced glucose output was not impaired by inhibition of prostanoid synthesis and function with the cyclooxygenase inhibitor indomethacin and the thromboxane receptor antagonist daltroban. These results indicate that HC specific defense reactions might be differently regulated under normal and inflammatory conditions as shown here for the indirect prostanoid-dependent or direct C5a-induced activation of hepatocellular glycogen phyosphorylase and glucose output in control or IL-6-treated rats, respectively. PMID- 10799913 TI - The inhibition of apoptosis in myositis and in normal muscle cells. AB - The mechanism of injury and death of muscle cells in the inflammatory myopathies (dermatomyositis, polymyositis, and inclusion body myositis) remains obscure. We and others have not detected apoptosis in the muscle biopsies from patients with myositis despite clear evidence of cell damage and loss. We provide evidence in this study that Fas ligand (FasL) as well as Fas is present on muscle cells and inflammatory cells in myositis biopsies: Fas is present on most muscle cells and lymphocytes, and FasL is present on degenerating muscle cells and many infiltrating mononuclear cells. The expression of both Fas and FasL in the inflamed tissue makes the absence of apoptosis more striking. To address the mechanisms of this resistance to classical apoptosis in muscle cells, we have investigated the expression of the antiapoptotic molecule FLICE (Fas-associated death domain-like IL-1-converting enzyme)-inhibitory protein (FLIP) in muscle biopsies of myositis patients and in cultured human skeletal muscle cells. Using laser capture microscopy, we have shown that FLIP is expressed in the muscle fibers and on infiltrating lymphocytes of myositis biopsies. Furthermore, we have shown that FLIP, but not Bcl-2, is expressed in cultured human skeletal muscle cells stimulated with proinflammatory cytokines, and inhibition of FLIP with antisense oligonucleotides promotes significant cleavage of poly(ADP-ribose) polymerase autoantigen, a sensitive indicator of apoptosis. These studies strongly suggest that the resistance of muscle to Fas-mediated apoptosis is due to the expression of FLIP in muscle cells in the inflammatory environment in myositis. PMID- 10799914 TI - Thymic and postthymic regulation of diabetogenic CD8 T cell development in TCR transgenic nonobese diabetic (NOD) mice. AB - Natural development of diabetes in nonobese diabetic (NOD) mice requires both CD4 and CD8 T cells. Transgenic NOD mice carrying alphabeta TCR genes from a class I MHC (Kd)-restricted, pancreatic beta cell Ag-specific T cell clone develop diabetes significantly faster than nontransgenic NOD mice. In these TCR transgenic mice, a large fraction of T cells express both transgene derived and endogenous TCR beta chains. Only T cells expressing two TCR showed reactivity to the islet Ag. Development of diabetogenic T cells is inhibited in mice with no endogenous TCR expression due to the SCID mutation. These results demonstrate that the expression of two TCRs is necessary for the autoreactive diabetogenic T cells to escape thymic negative selection in the NOD mouse. Further analysis with MHC congenic NOD mice revealed that diabetes development in the class I MHC restricted islet Ag-specific TCR transgenic mice is still dependent on the presence of the homozygosity of the NOD MHC class II I-Ag7. PMID- 10799915 TI - Human autoreactive CD4+ T cells from naive CD45RA+ and memory CD45RO+ subsets differ with respect to epitope specificity and functional antigen avidity. AB - T cells with specificity for self-Ags are normally present in the peripheral blood, and, upon activation, may target tissue Ags and become involved in the pathogenesis of autoimmune processes. In multiple sclerosis, a demyelinating disease of the CNS, it is postulated that inflammatory damage is initiated by CD4+ T cells reactive to myelin Ags. To investigate the potential naive vs memory origin of circulating myelin-reactive cells, we have generated myelin basic protein (MBP)- and tetanus toxoid-specific T cell clones from CD45RA+/RO- and CD45RO+/RA- CD4+ T cell subsets from the peripheral blood of multiple sclerosis patients and controls. Our results show that 1) the response to MBP, different from that to TT, predominantly emerges from the CD45RA+ subset; 2) the reactivity to immunodominant MBP epitopes mostly resides in the CD45RA+ subset; 3) in each individual, the recognition of single MBP epitopes is skewed to either subset, with no overlap in the Ag fine specificity; and 4) in spite of a lower expression of costimulatory and adhesion molecules, CD45RA+ subset-derived clones recognize epitopes with higher functional Ag avidity. These findings point to a central role of the naive CD45RA+ T cell subset as the source for immunodominant, potentially pathogenic effector CD4+ T cell responses in humans. PMID- 10799916 TI - Impact of HIV-1 infection on VH3 gene repertoire of naive human B cells. AB - B cells of the largest Ig variable heavy chain gene (VH) family, VH3, are reportedly decreased in patients with late stage HIV-1 disease. This deficit may contribute to their impaired responses to infections and vaccines. We confirmed that the VH3 family was underrepresented in serum IgM proteins, with a 45% decrease in patients with advanced HIV-1 disease. However, the proportion of VH3 within VH(1-6) IgM mRNA from peripheral B cells did not differ from that of control subjects (mean +/- SD, 57.1 +/- 9.7 vs 61.1 +/- 8. 7%). Similarly, within VH(1-6) IgD mRNA, which even more closely represents the unstimulated naive repertoire, the relative expression of VH3 mRNA was comparable in the two groups. Moreover, the frequency of individual genes within the VH3 family for IgD, particularly genes which encode putative HIV-1 gp120 binding sites, also was normal in HIV-1-infected patients. However, VH3 family expression for IgG mRNA was significantly decreased (17%) and VH4 IgG was increased (33%) relative to other VH families in advanced HIV-1-infected patients. Thus, the changes in VH family expression were more readily apparent in previously activated IgG "memory" B cell populations and, likely, in cells actively producing IgM rather than in resting naive cells. The presence of a relatively normal naive VH3 IgM and IgD mRNA repertoire in resting cells supports the prospect that with proper stimulation, particularly in conjunction with effective antiviral therapy, vigorous humoral immune responses to infections and vaccines may be elicited in this high-risk population. PMID- 10799917 TI - Induction of an antigen-specific CTL response by a conformationally biased agonist of human C5a anaphylatoxin as a molecular adjuvant. AB - A conformationally biased decapeptide agonist of human C5a anaphylatoxin (YSFKPMPLaR) was used as a molecular adjuvant in stimulating an Ag-specific CTL response against murine P815S target cells expressing an Ld-restricted CTL epitope of the hepatitis B surface Ag (HBsAg). Groups of BALB/c mice (H-2d) were immunized with aqueous solutions of the HBsAg CTL epitopes (IPQSLDSWWTSL and IPQSLDSWWTSLRR); the C5a agonist (YSFKPMPLaR); the C5a agonist and HBsAg CTL epitopes admixed (IPQSLDSWWTSL and IPQSLDSWWTSLRR + YSFKPMPLaR); the C5a-active, HBsAg CTL epitope-C5a agonist constructs (IPQSLDSWWTSLYSFKPMPLaR, IPQSLDSWWTSLRRYSFKPMPLaR, and IPQSLDSWWTSLRVRRYSFPMPLaR); a C5a-inactive, reverse moiety construct (YSFKPMPLaRRRIPQSLDSWWTSL); and a C5a-attenuated, carboxyl terminal-blocked construct (IPQSLDSWWTSLRRYSFKPMPLaRG). Ag-specific CD8+ CTL responses were observed after the secondary boost in the absence of any added adjuvant only in mice that were immunized with C5a-active contructs, IPQSLDSWWTSLRRYSFKPMPLaR and IPQSLDSWWTSLRVRRYSFKPMPLaR. These two C5a-active immunogens contained potential subtilisin-sensitive linker sequences between the HBsAg CTL epitope and the C5a agonist; i.e., a double-Arg (RR) and a furin protease sensitive sequence (RVRR). The introduction of these potentially cleavable sequences may be a method of increasing the likelihood of liberating the CTL epitope from the C5a agonist by intracellular proteases, thereby facilitating entry of the epitope into Ag-processing pathways via an exogenous route. PMID- 10799918 TI - Repertoire requirements of CD4+ T cells that prevent spontaneous autoimmune encephalomyelitis. AB - Spontaneous experimental autoimmune encephalomyelitis arises in 100% of mice exclusively harboring myelin basic protein-specific T cells, and can be prevented by a single injection of CD4+ T cells obtained from normal donors. Given the powerful regulatory effect of the transferred T cells, we further investigated their properties, and, in particular, their repertoire requirements. Transfer of monoclonal OVA-specific CD4+ T cells did not confer protection from disease even when present at very high proportions (about 80% of total lymphocytes). Lack of protection was also evident after immunization of these animals with OVA, indicating that not just any postthymic CD4+ T cells has the potential to become regulatory. However, protection was conferred by cells bearing limited TCR diversity, including cells expressing a single Valpha4 TCR chain or cells lacking N nucleotides. We also investigated whether coexpression of the myelin basic protein-specific TCR with another TCR in a single cell would alter either pathogenesis or regulation. This was not the case, as myelin basic protein specific/OVA-specific recombinase activating gene-1-/- double TCR transgenic mice still developed experimental autoimmune encephalomyelitis spontaneously even after immunization with OVA. Based on this evidence, we conclude that CD4+ T regulatory cells do not express canonical TCRs and that the altered signaling properties brought about by coexpression of two TCRs are not sufficient for the generation of regulatory T cells. Instead, our results indicate that regulatory T cells belong to a population displaying wide TCR diversity, but in which TCR specificity is central to their protective function. PMID- 10799919 TI - Human dendritic cells transfected with RNA encoding prostate-specific antigen stimulate prostate-specific CTL responses in vitro. AB - Although immunological tolerance to self Ags represents an important mechanism to prevent normal tissue injury, there is growing evidence that tolerance to tumor Ags, which often represent normal peripherally expressed proteins, is not absolute and can be effectively reverted. Prostate-specific Ag (PSA) is a self Ag expressed by both normal and malignant prostatic epithelium, and therefore offers a unique opportunity to examine the ability of self Ags to serve as specific CTL targets. In this study, we investigated the efficacy of autologous dendritic cells (DC) transfected with mRNA encoding PSA to stimulate CTL against PSA Ags in vitro. Ag in form of RNA carries the advantage to encode multiple epitopes for many HLA alleles, thus permitting induction of CTL responses among many cancer patients independent of their HLA repertoire. In this study, we show that PSA mRNA-transfected DC were capable of stimulating primary CTL responses against PSA Ags in vitro. The PSA-specific CTL did not cross-react with kallikrein Ags, a protein, which shares significant homology with PSA, suggesting that harmful autoimmune toxicity may not represent a significant problem with this approach. PSA RNA-transfected DC generated from male or female healthy volunteers or from cancer patients were equally effective in stimulating PSA-specific CTL in vitro, implying that neither natural tolerance to PSA Ags nor tumor-mediated T cell anergy may represent major barriers for CTL generation against the self Ag PSA. This study provides a preclinical rationale for using PSA RNA-transfected DC in active or adoptive immunization protocols. PMID- 10799922 TI - Corrections PMID- 10799921 TI - Chlamydia pneumoniae inhibits apoptosis in human peripheral blood mononuclear cells through induction of IL-10. AB - Chlamydia pneumoniae is a common cause of pulmonary infection, with serum positivity in at least 50% of the general population. In this study, we report that human PBMCs exposed to C. pneumoniae are resistant to apoptosis induced by the potent photoactivated chemotherapeutic agents 8-methoxypsoralen and hypericin. In contrast, PBMCs treated with a heat-inactivated inoculum exhibit normal susceptibility to apoptosis. We also observed that human PBMCs are responsive to C. pneumoniae infection by secretion of key immune regulatory cytokines, including IL-12 and IL-10. While IL-12 may play an important role in limiting C. pneumoniae proliferation within cells, IL-10 serves an anti inflammatory function by down-regulating proinflammatory cytokines such as IL-12 and TNF-alpha. Depletion of endogenous IL-10, but not of IL-12, abolished the apoptosis resistance of C. pneumoniae-infected PBMCs. Furthermore, addition of exogenous IL-10, but not IL-12, significantly increased the resistance of control inoculum-treated PBMCs to photoactivated 8-methoxypsoralen- and hypericin-induced apoptosis. Therefore, we conclude that C. pneumoniae possesses an antiapoptotic mechanism. The resistance to apoptosis observed in PBMCs exposed to C. pneumoniae is due, at least partially, to the IL-10 induced during C. pneumoniae infection. PMID- 10799920 TI - Enhanced susceptibility to lupus contributed from the nonautoimmune C57BL/10, but not C57BL/6, genome. AB - Genes from New Zealand Black and New Zealand White mice have been implicated in the development of a disease similar to human systemic lupus erythematosus. In an attempt to define the MHC class II genes involved in disease, we previously studied similarly designed backcrosses of New Zealand Black mice with C57BL/6 (B6) mice transgenic for Ez genes or with C57BL/10 (B10) mice transgenic for Az genes. Although the transgenes showed no effect on the development of autoantibody production or lupus nephritis in either backcross, surprisingly, there was greatly increased expression of these disease traits in the backcrosses involving B10 compared with B6 mice. These studies therefore implicated genetic contributions in B10 vs B6 backgrounds, despite their 98% identity. A genome-wide linkage analysis uncovered a B10 locus on mid-chromosome 13, which enhanced nephritis and was strongly linked with the production of pathogenic retroviral gp70-anti-gp70 immune complexes when contributed by B10, but not B6, mice. The subsequent identification of a single marker polymorphic between B10 and B6, along with the extreme genetic similarity between the two strains in this region, is likely to permit expedited identification of the lupus-susceptibility gene from this nonautoimmune strain. PMID- 10799924 TI - [Optimization of the recovery of 99m Tc-ciprofloxacin on Sephadex]. PMID- 10799923 TI - [Value of scintigraphy with 99mTc-MIBI in the diagnosis and follow-up of Hodgkin's and non-Hodgkin's lymphomas]. AB - This work aimed to assess the utility of 99mTc-MIBI scintigraphy in the diagnosis and follow-up of lymphoma patients. Seventy-two patients with a previous diagnosis of lymphoma (41 with Hodgkin's and 31 with non-Hodgkin's disease) and a control group of 8 patients with palpable lymph nodes and a negative diagnosis of lymphoma based on the clinical examination, chest x-ray, lymphography, ultrasounds and biopsy were studied. Whole body scintigraphies were performed 20 min and 2 h after i.v. administration of 740 MBq of 99mTc-MIBI. The joint result of the already mentioned standard diagnostic tests was considered as the confirmation criterion. A sensitivity, specificity and accuracy of 96,2%, 100% and 97,4% respectively were obtained. Kappa coefficient was k = 0,94 +/- 0,11. CONCLUSION: Scintigraphy with 99mTc-MIBI is a useful tool for the diagnosis and follow-up of lymphomas regardless of their classification. PMID- 10799925 TI - [Neuroanatomy of the nuclear vestibular complex in the syrian hamster. Comparison with other mammals]. AB - The vestibular complex has been studied since the fifties. The general conclusion of research on many mammals is that the vestibular complex has four main vestibular nuclei and some less constantly associated neuronal groups. The general distribution of the four main vestibular nuclei in the hamster does not differ substantially from that of other mammals: humans, many primates, cat, opossom, rabbit, chinchilla, guinea pig, etc. Of the many associated groups that have been described, we clearly identified groups <>, <>, <>, <>, and the interstitial nucleus of the vestibular nucleus of Cajal. However, the latter seems to be less developed than in other mammals. We present results and a map of serial sections of the vestibular area showing the most characteristic anatomic relations with brainstem structures, and the most relevant cytomorphometric results compared with other mammals. PMID- 10799926 TI - [Benign positional paroxysmal vertigo in the general ear, nose, and throat clinic]. AB - Benign positional paroxysmal vertigo is a common disease which may be caused by abnormal movement of utricular debris in the posterior semicircular canal. It is diagnosed by the Dix-Hallpike positional maneuver eliciting vertigo and nystagmus. Treatment generally consists of physical exercises with the Epley or Semont maneuvers. We review 43 consecutive patients diagnosed as BPPV in the last year and treated with physical therapy (Epley maneuvers). Results were very good, with a cure rate of 88.37% (53.5% after a single maneuver). Physical therapy is an effective treatment for BPPV. PMID- 10799927 TI - [Descriptive study of infectious ear disease in relation to summer]. AB - A descriptive study was made of infectious ear disease (including diffuse otitis externa, otomycosis, acute-on-chronic otitis media, and superinfection of a radical mastoidectomy cavity) in relation to changes of weather and habits in summer. During the months of June, July, and August 1996, 179 patients were evaluated in the emergency room of the Alicante General University Hospital, Spain. Average patient age at presentation was 30.52 (+/- 20.08) years and 56% were men. The most frequent disease was diffuse otitis externa (78%) followed by acute-on-chronic otitis media (12%), otomycosis (8%), and superinfection of a radical mastoidectomy cavity (2%). The most frequently involved microorganisms were Pseudomonas aeruginosa in diffuse otitis externa, Aspergillus niger and Candida in otomycosis, Escherichia coli, Haemophilus influenzae, Proteus mirabilis, and Staphylococcus aureus in acute-on-chronic otitis media. Patients were treated by cleaning detritus and secretions, usually followed by topical antibiotics for a maximum period of one week. PMID- 10799928 TI - [Diagnosis of common causes of vertigo using a structured clinical history]. AB - The structured clinical history is the most sensitive test for diagnosing vertigo. Its diagnostic effectiveness on the first visit was analyzed and key signs and symptoms with high predictive value for common causes of vertigo were identified. One hundred outpatients who complained of dizziness or loss of balance were evaluated using a structured clinical interview. Each questionnaire was examined independently by three blinded investigators, who assigned a diagnosis and identified the elements of the history that figured most prominently in the diagnosis. The gold standard was defined as independent selection of the same diagnostic category by all three investigators. A first visit diagnosis was obtained in 40% of patients (95% confidence interval 30-50%): 38% women and 42% men. Causes included benign positional paroxysmal vertigo (BPPV, 13 patients), headache-associated vertigo (9), Meniere disease (7), cervical vertigo (3), psychiatric dizziness (2), post-traumatic vertigo (2), vertebro-basilar transient ischemic attack (1), vestibular neuritis (1), convulsive seizure (1), and presyncope (1). The best predictors of BPPV were the precipitating mechanism (specificity [SP] 100%), positional nystagmus (sensitivity [SE] 90%, SP 63%), and the Dix-Hallpike test (SE 82%, SP 71%). Elements predictive of headache-associated vertigo were duration of the attack (minutes) and a personal history of headache (both, SP 100%). Other predictors were facial hypoesthesia (SE 92%, SP 47%) and associated neurological disease (SE 82%, SP 58%). PMID- 10799929 TI - [Early diagnosis of childhood deafness: detection protocol in high-risk neonates]. AB - The protocol developed at our hospital for auditory assessment of children with risk factors for deafness is presented. The results obtained with tympanometry and auditory brainstem response (ABR) audiometry from October 1994 to March 1998 were analyzed. In 122 children, 17 cases (14%) of severe-to-profound bilateral deafness were found. Only 50% of these cases were associated with risk factors present at birth. The average age at time of diagnosis of deafness was over 3 years. In view of these results, we recommend auditory screening for all newborns as the only effective procedure for early detection of childhood deafness. PMID- 10799930 TI - [A new sclerosant for the treatment of Rendu-Osler-Weber disease]. AB - We report the cases of two women with Rendu-Osler-Weber disease and severe epistaxis. After several procedures failed (electrocoagulation, vascular clamping, dermoplasty, embolization), submucosal infiltration of a sclerosant was performed using the new product etoxysclerol. Epistaxis remained under control after two years of monitoring. PMID- 10799931 TI - [Predictive value of the Muller maneuver in obstructive sleep apnea syndrome]. AB - Dynamic changes in the upper airway during sleep have been identified in patients with obstructive sleep apnea syndrome (OSAS). Fiberoptic nasopharyngoscopy with the Muller maneuver (FNMM) has been advocated as a practical, clinically useful, preoperative dynamic diagnostic method for assessing the site of collapse. The value of FNMM in predicting response to uvulopalatopharyngoplasty (UPPP) was assessed in patients with obstructive sleep apnea syndrome. Results indicate that preoperative FNMM can reliably enhance surgical success. PMID- 10799932 TI - [Laryngeal fibroendoscopic surgery. Our experience]. AB - Since 1992, when our department acquired a flexible fiberscope with a working channel, we have used it in surgery on the upper respiratory and digestive tract. We call our technique laryngeal fiberscopic surgery. We report the results of four years of experience in this field. A total of 368 surgical interventions were performed, almost half to remove benign lesions of the vocal cords (polyps, nodules, etc). This technique is a major advance in our field of study. Its main advantages are avoidance of general anesthesia and hospitalization, reduction of surgical risk, and savings of time and money. We consider it to be a useful alternative to traditional direct laryngoscopy (Kleinsasser technique) for many applications. PMID- 10799933 TI - [Spectrographic study of voice disorders: subharmonics]. AB - The presence of subharmonics in abnormal adult voices (polyps, polypoid degeneration, plicae ventricularis) was investigated. Subharmonics are low frequency modulations located between two consecutive harmonics in the spectrum. Spectrum analysis was performed on 115 abnormal voices, of which 36 (31%) had subharmonics. Thirteen of the 36 voices with subharmonics had normal jitter and shimmer, so the rough voice was characterized acoustically not only by jitter and shimmer but by the presence of subharmonics in the spectrum. Subharmonics may correspond to a qualitative change in the behavior of the vibratory system. PMID- 10799934 TI - [Radiotherapy in glottic T1 carcinoma. Local control and survival]. AB - Between January 1980 and April 1995, 57 treatment-naive patients diagnosed as glottic epidermoid T1 carcinoma were treated in the ear, nose and throat department of Juan Canalejo Hospital of La Coruna, Spain. Seventy-nine percent (79%) (45/57) had T1a lesions and 21% (12/57) had T1b. All patients were treated by the radiotherapy department of the Regional Oncological Center with Co60 at doses ranging from 50 Gy to 70 Gy. Local control was achieved in 74.5% (43/57). Cause-specific survival was 96% at 3, 5, and 10 years. The larynx was preserved in 96.5% (55/57). The local recurrence rate was significantly lower for T1a (16.8%) than for T1b (57.8%) (p < 0.05). No significant differences were found in local control or survival with different dosing regimens. PMID- 10799935 TI - [Tularemia in the differential diagnosis of cervical lymph node enlargement. An outbreak of tularemia in Castilla-Leon, Spain]. AB - The presence of enlarged cervical lymph nodes is a diagnostic challenge in disease of the head and neck. Lymph node enlargement may be secondary to local or general infectious disease, to non-infectious systemic disease, or to lymphatic metastasis of tumoral processes. Among the many infectious processes that originate cervical lymph nodes is tularemia. This disease is uncommon in Spain, but was unusually frequent in Castilla-Leon in the last months of 1997, with 136 cases diagnosed in our hospital, 13 accompanied by lymph node enlargement of the head and neck. This article shows that tularemia should be considered in the differential diagnosis of cervical lymph node enlargement in our region. PMID- 10799936 TI - [Hernia of the external ear canal of uncommon origin]. AB - The temporo-mandibular joint is directly related to the anterior wall of the external ear canal. Therefore, defects in this wall may lead to herniation of the soft tissues of the temporo-mandibular joint into the external ear canal. Such herniation usually is secondary to external injuries, iatrogenic trauma, neoplasms, and so on, but it may also be congenital. We report a well-documented case of temporo-mandibular joint herniation into the external ear canal of probable congenital origin. PMID- 10799937 TI - [Mediofacial degloving in extramedullary plasmacytoma with orbital invasion]. AB - Extramedullary solitary plasmacytoma is a rare plasmatic cell tumor occurring in the head and neck. We report the case of a patient with a mass that arose from the sphenoidal base, extended toward the right paranasal sinuses, and invaded the orbit. The clinical characteristics, diagnostic criteria, and therapeutic problems are discussed and reviewed in the literature, with special attention to an alternative therapeutic approach, midfacial degloving. PMID- 10799938 TI - [Ethmoidal metastasis of renal adenocarcinoma]. AB - The sinonasal tract and, more specifically, the ethmoid sinus, are uncommon sites for metastatic tumors. Fewer than 100 cases have been reported in the world literature. We report the case of a 65-year-ol woman who underwent nephrectomy and adrenalectomy for renal cell carcinoma (hypernephroma) with adrenal metastasis. Four months later the patient was seen for repeated epistaxis and nasal obstruction, which was diagnosed as ethmoidal metastasis of renal adenocarcinoma in a second biopsy. The second metastasis was removed by lateral rhinotomy but recurred six months later. Palliative surgery was performed and the patient died eight months later from brain metastases. Therapeutic strategies for metastatic nasal tumors are reviewed. PMID- 10799940 TI - [Leiomyosarcoma of the larynx]. AB - A patient with leiomyosarcoma of the larynx met diagnostic criteria for radiation induced malignancy. In a review of the literature, only 27 leiomyosarcomas of the larynx were found, none radiation-induced. The histological features and diagnosis and treatment are analyzed. PMID- 10799941 TI - [Inflammatory pseudotumor of the lymph nodes. A case report]. AB - Inflammatory pseudotumor of the lymph nodes is a little-known condition characterized mainly by cervical lymph node enlargement, hematological manifestations, and major constitutional symptoms in most patients. The diagnosis is histological, after surgical removal of an involved lymph node. Histologically, stromal proliferation and neovascularity are present, as well as lymphocytic infiltration. This disease is considered benign because of its tendency toward spontaneous resolution, even though symptoms are usually intense. A male patient presented cervical lymph node enlargement, constitutional syndrome, and hematological manifestations, including oscillating thrombocytopenia, leukopenia, and increased ESR. The diagnosis was inflammatory pseudotumor of the lymph nodes, a rare disease. The evolution is torpid but the prognosis is good. This disease should be considered in the differential diagnosis of tumors of the neck region. PMID- 10799939 TI - [Lingual actinomycosis: diagnostic problems. Review of the literature]. AB - Actinomycosis of the cervicofacial area is a rare infection with a difficult clinical diagnosis. We report a case of lingual actinomycosis which was diagnosed after a long remission with empirical antibiotic treatment, two years after onset. The clinical characteristics, differential diagnosis, and therapeutic options in this unusual infection are discussed. PMID- 10799942 TI - [Myasthenia gravis. Otorhinolaryngological considerations]. AB - Myasthenia gravis is a motor neuron disease caused by the presence of antibodies against acetylcholine receptors that interfere with proper functioning of the neuromuscular junction. Twenty percent of patients show bulbar involvement as the first indication of disease, with symptoms such as rhinolalia, dysphagia or phonasthenia. We report the cases of five patients for which our intervention was requested. We were involved in capacities ranging from the interpretation of the first symptom of disease to assessment of surgical possibilities for the treatment of chronic aspiration and severe respiratory symptoms in patients with major dysphagia. We review the scant bibliography published in the last five years. PMID- 10799943 TI - Anisakis simplex: a cause of intestinal pseudo-obstruction. AB - OBJECTIVE: the ingestion of Anisakis simplex larvae may lead to the appearance of gastrointestinal symptoms. However, the number of reported cases of parasitization by Anisakis in Spain is lower than would be expected in a country with the second-highest fish consumption per inhabitant in the world, particularly since fish is often eaten raw or only slightly cooked. We suggest that the incidence of anisakiasis in Spain would be higher if complementary studies were used in all patients suspected of having anisakiasis. METHODS: we studied 6 patients with a diagnosis of intestinal obstruction who frequently ate fish. Skin prick tests with seafood, inhalant allergen and Anisakis extracts were done. Total and specific IgE against Anisakis larvae were tested with a CAP system radioimmunoassay and immunoblot assays. Oral challenge tests with frozen larvae were also used. RESULTS: a positive skin prick result and high levels of total and specific IgE were found in all patients. The results of immunoblot assays for IgE did not show a consistent pattern, but a group of several low (14 18 kDa) and intermediate molecular weight antigens (30-50 kDa) were found in all patients. All patients tolerated the oral challenge test well. CONCLUSIONS: in our patients with intestinal pseudo-obstruction and a history of frequent fish eating, the clinical and laboratory findings were suggestive of parasitization by Anisakis simplex larvae as the cause of the obstruction. Such complementary studies should be used whenever there is a suspicion of anisakiasis. The results of the oral provocation test show that the intake of dead larvae does not induce clinical parasitization. PMID- 10799944 TI - Rapid method for the detection of anti-HCV antibodies in patients with chronic hepatitis C. AB - OBJECTIVE: to evaluate the usefulness of a simple, rapid, qualitative technique (MedMira Rapid Test) to detect antibodies against hepatitis C virus (HCV) and compare this approach with an immunometric technique in patients with chronic hepatitis C infected with different genotypes. METHODS: anti-HCV antibodies were determined with the MedMira rapid technique and an immunometric method in samples from 138 patients with chronic hepatitis C infected with different HCV genotypes, and in 50 samples from healthy individuals. RESULTS: the MedMira rapid technique detected anti-HCV antibodies in 135 (98%) of 138 serum samples from patients with chronic hepatitis C, whereas the immunometric method gave positive results in all 138 samples. Three of the 138 anti-HCV-positive samples identified with the immunometric method and confirmed by inmunoblotting were repeatedly negative with the MedMira rapid technique. Two of these samples were genotype 1 and the third was not genotyped. All samples from the control group were negative for anti-HCV antibodies by both methods. The sensitivity and specificity of the MedMira rapid technique relative to the immunometric technique were 98% and 100% respectively. CONCLUSION: the MedMira rapid technique is a quick, specific and sensitive method that is easy to use by nonspecialized personnel, and is a good alternative to other, slower methods for the diagnosis of chronic hepatitis C. PMID- 10799945 TI - Role of manometry, defecography and anal endosonography in the evaluation of colorectal disorders. AB - OBJECTIVE: altered motility or anatomy of the rectum, anus and perineal floor may lead to symptoms which are unresponsive to routine therapeutic approaches. These disturbances usually lead to constipation, fecal incontinence, or both. Different tests and techniques for evaluating anorectal and perineal disorders, developed in the last two decades, make a better understanding of these disorders possible. This study was designed to evaluate the diagnostic benefits of combining manometry, defecography and anal endosonography in the assessment of patients with anorectal disorders. METHODS: twenty-five children with constipation (with or without soiling), incontinence and/or prolapse underwent anal manometry, defecography and anal endosonography. Group A consisted of 9 children with fecal incontinence, group B consisted of 10 children with constipation with soiling, and group C comprised 6 children with constipation without soiling. RESULTS: in group A resting incontinence was associated with a hypotonic external sphincter in 4 out of 9 patients, 2 of whom had internal anal sphincter thinning. In group B resting incontinence was associated with a hypotonic external sphincter in 8 out of 10 patients, 6 of whom had internal anal sphincter thinning. In group C these associations were not seen in any of the patients. CONCLUSIONS: barium enema is not sufficient for an accurate diagnosis of anorectal disorders. No single test is capable of revealing the type of disease. Anal manometry, defecography and endosonography are complementary procedures in the assessment of this group of disorders. This new approach will improve our knowledge of the pathogenesis of these disorders in children. However, further studies are needed to obtain conclusive evidence. PMID- 10799947 TI - [Atypical premiere of a metastatic gastric adenocarcinoma] [In Process Citation] PMID- 10799946 TI - Clinical relevance of Helicobacter pylori CagA-positive strains: gastroduodenal peptic lesions marker. AB - OBJECTIVES: peptic ulcer is characterized by its recurrent nature, which necessitates maintenance treatment in most patients. But this natural history can be changed in patients with peptic ulcer associated to Helicobacter pylori, as shown by the low rates of recurrence and decreased hemorrhagic recidivism associated with this infection. Whether CagA or VacA strains are associated with a greater risk of peptic ulcer is controversial. This study was designed to examine endoscopic findings and their relation with H. pylori phenotype (CagA or VacA). METHODS: 106 selected dyspeptic patients underwent upper gastrointestinal tract endoscopic examination between September 1996 and May 1997 [69 with H. pylori (Hp) and 37 without this infection]. Endoscopic findings were classified as gastric ulcer (GU), duodenal ulcer (DU), gastric erosions (GE), duodenitis (Du), chronic gastritis (CG) and normal mucosa (NM). Hp phenotype was analyzed with a western blot test. RESULTS: 75% of H. pylori strains were CagA-positive and 54.2% were VacA-positive. 82.4% of the cases of DU were associated with a CagA+ phenotype, but the association was not statistically significant. Otherwise 100% of gastric ulcers were associated with CagA+ strains (p < 0.005). VacA phenotype was not associated with any particular endoscopic finding. Peptic ulcer (DU or GU) was also associated with the CagA+ phenotype (p < 0.05). CONCLUSIONS: the CagA+ H. pylori phenotype seems to be a peptic lesion marker, but was more frequently related with GU than with DU in our sample of Spanish patients. PMID- 10799949 TI - [Hydatic disease of the pancreas] [In Process Citation] PMID- 10799948 TI - [Portal and mesenteric thrombosis and protein <> deficiency] [In Process Citation] PMID- 10799950 TI - [Transmural hematoma of the rectum complicating anticoagulant therapy] [In Process Citation] PMID- 10799951 TI - [Resolutive fibrinolysis in a case of acute mesenteric ischaemia] [In Process Citation] PMID- 10799967 TI - Mapping genes for polygenic disorders: considerations for study design in the complex trait of inflammatory bowel disease. AB - While the methodology for the mapping of Mendelian disorders is well established, the practical and theoretical steps required for successful gene identification in a complex trait are still difficult to predict. A number of analytical models and simulations based on repetitive drawings from predefined statistical distributions are available. To supplement these analytical models, we developed an integrated simulation approach by directly simulating entire populations under a disease model based on epidemiological data. Random mating, nonoverlapping populations and the absence of differential fitness were assumed. Samples were drawn from these homogeneous and heterogeneous populations and analyzed with established analysis tools. We investigated the properties of linkage and association studies in inflammatory bowel disease - modeled as a six-locus polygenic disorder - as an example of this approach. In nonparametric linkage studies, lod scores varied widely, with the median required sample size depending on the locus-specific relative sibling risk. A fine mapping resolution <4 cM was found to require nonparametric lod scores >10. Family-based association studies (TDT test) and case-control studies showed a similar sensitivity and can identify risk loci in populations with moderate levels of linkage disequilibrium in sample sizes of 500-800 triplets. Case-control association studies were prone to false positive results if applied in heterogeneous populations, with the false-positive rate increasing with sample size because population heterogeneity is detected with increasing power. PMID- 10799968 TI - Fine mapping of the human biotinidase gene and haplotype analysis of five common mutations. AB - Biotinidase deficiency is an autosomal recessive defect in the recycling of biotin that can lead to a variety of neurologic and cutaneous symptoms. The disease can be prevented or effectively treated with exogenous biotin. The biotinidase locus (BTD) has been maped to 3p25 by in situ hybridization. The gene has been cloned, the coding region sequenced, the genomic organization determined, and a spectrum of mutations has been characterized in more than 90 individuals with profound or partial biotinidase deficiency. We have conducted haplotype analysis of 10 consanguineous and 39 nonconsanguineous probands from the United States and 8 consanguineous probands from Turkey to localize BTD with respect to polymorphic markers on 3p and to investigate the origins of five common mutations. The inbred probands were homozygous for overlapping regions of 3p ranging in size from 1.1 to 80 cM which were flanked most narrowly by D3S1259 and D3S1293. Radiation hybrids and haplotype analysis of markers within this region suggest that BTD is located within a 0.1-cM region flanked by D3S3510 and D3S1286. The radiation hybrid data suggest that the BTD gene is oriented 5' to 3' between the centromere and the 3p telomere. Association studies indicate that the gene is closer to a third locus D3S3613 than D3S3510, two markers which cannot be resolved by existing linkage data. The BTD locus and D3S3613 must therefore lie between D3S3510 and D3S1286. Comparison of haplotypes reveals evidence for possible founder effects for four of the five common mutations. PMID- 10799970 TI - alpha-thalassemia in Bantu population from Congo-Brazzaville: its interaction with sickle cell anemia. AB - Deletional alpha(+)-thalassemia (-alpha(3.7)) was investigated in four groups of unrelated individuals from the Bantu population (newborns, normal adults, sickle cells trait carriers, sickle cell anemia patients) of Brazzaville, Congo. The frequency of the (-alpha(3.7)) chromosome was similar between newborns (f = 0.40) and adult subjects (f = 0.36), and between sicklers and nonsickler subjects. The frequency of the (-alpha(3.7)) chromosome in sickle cell anemia patients (SS patients) did not change when age was stratified. The hematological characteristics of SS patients with (-alpha/alphaalpha, -alpha/-alpha) and without (alphaalpha/alphaalpha) alpha(+)-thalassemia were similar to those reported in Jamaican and US sickle cell anemia patients. alpha(+)-Thalassemia had an effect on the percentage of hemoglobin S in sickle cell trait carriers. Thus, the high frequency of alpha(+)-thalassemia in the Congolese population presumably results from this disorder having a selective advantage favoring survival. However, the frequency of alpha(+)-thalassemia was not affected by age. Although in this selective tropical environment, alpha(+)-thalassemia as elsewhere markedly affects the hematological characteristics of sickle cell anemia patients, however our data provide no evidence that alpha(+)-thalassemia increases survival of SS patients. PMID- 10799969 TI - Allele-specific variation in the gene copy number of human cytosine 5 methyltransferase. AB - Previously, we have identified two alternate allelic forms of cytosine 5 methyltransferase, 5-MT I and 5-MT II, specified by polymorphic fragments of 1.5 and 1.1 kb, respectively. In the presence study, a 0.8-kb genomic probe was prepared which was confirmed to be included within the polymorphic fragments. The 0. 8-kb probe hybridised with greater intensity to the 1.1-kb fragment than the 1.5-kb fragment. Densitometric analysis indicated that there is 1 copy of 5-MT associated with 5-MT I, whereas there may be 1-4 copies of the gene associated with the 5-MT II allele. Segregation studies demonstrated that the multiple copies of 5-MT II are inherited in a Mendelian fashion. These results allow novel approaches to investigating the underlying mechanisms of cytosine methylation and gene duplication. PMID- 10799971 TI - Effects of misspecification of allele frequencies on the type I error rate of model-free linkage analysis. AB - Linkage analyses of simulated quantitative trait data were performed using the Haseman-Elston (H-E) sib pair regression test to investigate the effects of inaccurate allele frequency estimates on the type I error rates of this test. Computer simulations generating a quantitative trait in nuclear families were performed using GASP [1]. Assuming no linkage, several data sets were simulated; they differed in marker allele numbers and frequencies, number of sib pairs and number of sibships. Each set of simulated data was analyzed using (1) all parental marker data, (2) half of the parental marker data, and (3) no parental marker data, using both correct and incorrect allele frequencies in the latter 2 cases. The H-E sib pair linkage method was found to be robust to misspecification of marker allele frequencies regardless of the number of alleles. PMID- 10799972 TI - Model-free analysis and permutation tests for allelic associations. AB - In this short report, we address some practical problems in performing likelihood based allelic association analysis of case-control data. Model-free statistics are proposed and their properties assessed by simulation, and procedures based on permutation tests are described for marker-marker as well as marker-disease associations. A memory-efficient algorithm is developed which enables several highly polymorphic markers to be analysed. PMID- 10799974 TI - Cystic fibrosis mutations: report from the French Registry. The Clinical Centers of the CF. AB - Data from 2,666 patients with cystic fibrosis (CF) born in France, submitted during the period of 1992-1996 to the French registry for CF, were used to describe the different mutations, their frequency and their regional distribution. A total of 5,332 CF chromosomes have been analyzed, demonstrating 229 different mutations and accounting for 87% of CF genes in the French population. DeltaF508 is the most common mutation at 67.9% of CF mutations, followed by G542X (2.5%), N1303K (2.0%), 1717-1G-->A (1.2%), R553X (0.8%) and G551D (0.7%). The data show a clear geographical variation in the distribution of many of the mutations. Given the geographical heterogeneity of these mutations, carrier screening does not appear to be feasible in most French regions. PMID- 10799973 TI - A novel Q562X mutation identified in the hMLH1 gene in a Slovenian patient with hereditary nonpolyposis colorectal cancer. PMID- 10799975 TI - A new inherited interstitial deletion of the distal long arm of chromosome 4. AB - A 12-year-old boy showed mild dysmorphic features, late presentation of learning difficulties and behaviour problems, obesity, breast hypertrophy and bilateral slipped capital femoral epiphysis. His mother also has mild dysmorphic features, obesity, and a similar history of late presentation of learning difficulties and behaviour problems. Cytogenetic analysis demonstrated an inherited distal long arm deletion of one chromosome 4. The boy's karyotype was interpreted as 46,XY,del(4)(q32 q33)mat and the mother's karyotype as 46,XX,del(4)(q32 q33). This is the second report of an inherited distal 4q deletion and the first report of interstitial chromosome 4 deletion involving q32 q33 segments. PMID- 10799976 TI - The position of the LysN epsilon H2-grafted antigens along the sequential oligopeptide carrier, Ac-(Aib-Lys-Aib-Gly)n (SOCn-II), influences the antibody recognition: application to the Sm main autoimmune epitope. AB - A sequential oligopeptide carrier of antigenic peptides is presented, incorporating two Aib residues in each repetitive moiety: Ac-(Aib-Lys-Aib-Gly)(n) (SOC(n) -II; n = 2-4). The conformational study, by (1)H-nmr, CD, and Fourier transform ir spectroscopy, indicated that the SOC(n) -II carrier displays a pronounced 3(10)-helix, compared to the Ac-(Lys-Aib-Gly)(n) (SOC(n) -I) carrier of the same approximately backbone length, previously reported. One of the dominant autoimmune epitopes of the Sm and U1RNP cellular components, the PPGMRPP sequence, was coupled to the Lys-N(epsilon)H(2) groups of the SOC(n) -II carrier and used as antigenic substrate for detecting anti-Sm/U1RNP autoantibodies in ELISA assays. Anti-Sm antibodies are highly specific for systemic lupus erythematosus, while anti-U1RNP are specific for mixed connective tissue disease. The anti-(PPGMRPP)(5)-SOC(n) -II ELISA was compared with the anti-(PPGMRPP)(n) SOC(n) -I ELISA, provided that both antigenic substrates possess the same amount of the epitope replicates. The significance of the lysine positions along the oligopeptide backbone of the carrier for a favorable antibody recognition of the anchored antigens is also examined. PMID- 10799977 TI - A conformational study of the xyloglucan oligomer, XXXG, by NMR spectroscopy and molecular modeling. AB - A structural study of the XXXG xyloglucan heptasaccharide (X = alpha-D-Xylp(1 --> 6)-beta-D-Glcp and G = beta-D-Glcp) isolated from apple fruit has been undertaken with nmr and molecular mechanics methods. Quantitative 400 MHz nmr data including nuclear Overhauser effect spectroscopy (NOESY) volumes were recorded at both 6 and 20 degrees C. In spite of severe overlapping of resonances, it was possible to estimate summed NOEs for the majority of the anomeric and glucosyl methylene protons. An ensemble-average population of preferred geometries has been established with the CICADA conformational searching algorithm associated with the MM3 force field. Comparison of the theoretical data obtained by back calculation of the NOESY volumes from the ensemble-average distance matrix program and motional models based on the Stokes-Einstein-Debye relation satisfactorily reproduce the experimental data. Conformational averaging about the mainchain glycosidic linkages includes both the syn and anti conformers and a minor gauche-gauche population is highly probable. The theoretical data overestimate the syn preference of the Glc(c) --> Glc(b) linkage as well as the Glc(c) GT rotamer population. Finally, both the motional models and the conformational search indicate a fairly rigid backbone and greater flexiblity for the xylose side chains. PMID- 10799978 TI - Amphipathic structure of theonellapeptolide-Id, a hydrophobic tridecapeptide lactone from the Okinawa marine sponge Theonella swinhoei. AB - Theonellapeptolide-Id (TNLP), a cyclic tridecapeptide lactone, was crystallized from dimethylformamide-water solution. In the asymmetric unit, two peptide molecules were combined with solvent molecules, and the total molecular weight was over 3000 Dalton. The crystal structure including solvent molecules was finally determined at 0.80 A resolution using synchrotron radiation. The conformations of two independent molecules were similar to each other and were also similar to the previously reported structure (Doi, Ishida, Kobayashi, Deschamps and Flippen-Anderson, 1999, Acta Crystallogr Sect C, 55, 796-798). About 13 hydrated water molecules were found at disordered 19 sites; they were located at a certain region to avoid contact with aliphatic side-chains of peptolide in the crystal. The spatial disposition of the solvent molecules and peptides subsequently caused the formation of the amphipathic layer. PMID- 10799979 TI - The rotating frame nuclear Overhauser effect spectroscopy experiment as an aid to determining solution structures of DNA oligomers. AB - Important intrinsic characteristics of the rotating frame nuclear Overhauser effect spectroscopy (ROESY) experiment were found to be advantageous in DNA solution structure determination. In a ROESY experiment, the different mechanisms of relaxation result in different signs of cross peaks, enabling a clear distinction between H2' resonances and H2" resonances of the DNA sugar backbone. This method is of particular importance in crowded spectra, for purine resonances whose H2', H2" protons typically resonate closely, as well as in conditions where line broadening makes coupling constants in a correlated spectroscopy experiment impossible to determine. By observing the signs of cross peaks in the base proton to H2', H2" sugar proton region, the ROESY spectrum can be used to distinguish A form, B-form, and Z-form DNA. PMID- 10799980 TI - Structural basis for the biological activity of dendrotoxin-I, a potent potassium channel blocker. AB - A topochemical model to explain the biological activity of dendrotoxin-I (DTX-I), a potent blocker for potassium channels, was developed by searching common spatial arrangements of functionally important residues between DTX-I, alpha dendrotoxin, dendrotoxin-K, BgK, ShK, and charybdotoxin. The first three are structurally and functionally related to one another, and specifically target to Kv1 type potassium channels. The last three are structurally unrelated to the first three but have the ability to displace (125)I-labeled dendrotoxins on the same types of potassium channels. In order to obtain the correct electronic surface potential, thought to be crucial for the DTX-I function, we determined the three-dimensional solution structure of DTX-I by nmr spectroscopy using its correct amino acid sequence recently determined by our group. The most interesting characteristic of our model is that DTX-I has two binding sites to potassium channels: one is the cationic domain made up of Lys residues at positions 5 in the 3(10)-helix, 28 and 29 in the beta-turn, and the other is the Lys19/Tyr17/Trp37 triad located in the antiprotease domain. The cationic domain and the triad are located at the opposite sides of the molecular structure and are separated by about 25 A between Lys29 Calpha and Tyr17 Calpha. The functional triad is characterized by three distances, d(1) approximately 7.5 A (Lys19 Calpha the center of the Tyr17 aromatic ring), d(2) approximately 8.1 A (Lys19 Calpha the center of the 6-membered ring of the Trp37 indole group), and d(3) approximately 7. 3 A (the center of the Tyr17 aromatic ring-the center of the 6 membered ring of the Trp37 indole group). This model should aid in the pharmaceutical design of peptide and nonpeptide drugs with potassium channel blocking potencies, as well as in understanding of the physiology, pharmacology, biochemistry, and structure-function analysis of potassium channels. PMID- 10799981 TI - Applicability of broken-rodlike chain model to conformational analysis of polypeptide chain. AB - In order to check the applicability of the broken-rodlike (BR) chain model, consisting of several rods alternatively joined by flexible random coils, to the conformational analysis of a polypeptide chain in the helix-to-coil transition regions, two relations predicted by the Zimm and Bragg theory and the method with the BR chain model are compared. It is shown that, despite a clear difference between the models employed in the two methods, they give substantially identical results in both probability P(j) that a helical residue is in a helical sequence j units long and averaged helical fraction dependence of the mean-squared radius of gyration. Thus the use of the method with the BR chain model in the conformational analysis of a polypeptide chain could be rationalized, at least, with the same degree of approximation as is assumed in the Zimm and Bragg theory. Using the scattering function for the BR chain model, averaged helical-sequence lengths are evaluated for partially ionized poly(L-glutamic acid) (PGA) in added salt aqueous solution and nonionized PGA in N-methylacetamide, both in a helical state. As a result, it is shown that the length in the latter molecule is approximately tenfold longer than that in the former one. PMID- 10799982 TI - Analysis of a local fitness landscape with a model of the rough Mt. Fuji-type landscape: application to prolyl endopeptidase and thermolysin. AB - A method of analysis of a local fitness landscape for a current biopolymer is presented. Based on the assumption of additivity of mutational effects in the biopolymer, we assigned a site-fitness to each residue at each site. The assigned values of site-fitnesses were obtained by the least-squares method to minimize discrepancies between experimental fitnesses and theoretical ones. As test cases, we analyzed a section of a local landscape for the thermostability of prolyl endopeptidase and that for the enzymatic activity of thermolysin. These sections were proved to be of the rough Mt. Fuji-type with straight theta values of larger than 1.0, where straight theta is defined as the ratio of the "mean slope" to the "degree of roughness" on the fitness surface. Furthermore, we theoretically explained discrepancies between the fitnesses of multiple mutants and those predicted based on strict additivity of the component mutations by using a model of the rough Mt. Fuji-type landscape. According to this model, the discrepancies depend on the local landscape property (such as the straight theta value) and the location of the wild type on the landscape and the mean change in fitness by the component mutations. Our results suggest that this model may provide a good approximation of real sections of local landscapes for current biopolymers phenomenologically. PMID- 10799983 TI - Power consumption in shaking flasks on rotary shaking machines: I. Power consumption measurement in unbaffled flasks at low liquid viscosity. AB - In this first article of a series a new method is introduced that enables the accurate determination of the power consumption in a shaking flask. The method is based on torque measurements in the drive and appropriate compensation of the friction losses. The results for unbaffled shaking flasks at low viscosities are presented after varying shaking frequency, flask size, filling volume, shaking diameter, and surface quality (hydrophilic and hydrophobic) of the inner flask walls. The order of magnitude of the values of power consumption in shaking flasks is equal to, or even higher than, the values typical for agitated tank bioreactors. A physically based model equation for shaking flasks is derived that introduces a modified power number and a resulting constant as the only fitting parameter. With this equation, the measured results are correlated with sufficient accuracy. For the first time, comprehensive data for the power consumption in unbaffled shaking flasks at low viscosity is available, giving a detailed picture of the influences of the different variables. PMID- 10799984 TI - Power consumption in shaking flasks on rotary shaking machines: II. Nondimensional description of specific power consumption and flow regimes in unbaffled flasks at elevated liquid viscosity. AB - This article is the second part of a series presenting and modeling the hydrodynamics and specific power consumption in shaking flasks on rotary (orbital) shaking machines. In part I, a new method was introduced that enables the accurate determination of the specific power consumption in shaking flasks. The method was first applied to investigate unbaffled flasks with a nominal volume of < or =1 L at low viscosity. In part II, the results for the specific power consumption of unbaffled shaking flasks at elevated viscosities are investigated after varying shaking frequency, flask size, filling volume, and shaking diameter. The theory introduced in part I is extended to liquids of elevated viscosities using nondimensional equations. With these results, the specific power consumption in unbaffled shaking flasks can now be fully described. For the first time, the phenomenon of the liquid being "out of phase" is observed and described. This occurs at certain operating conditions and is characterized by an increasing amount of liquid not following the movement of the shaking table, thus reducing the specific power consumption. This, of course, has much relevance for practical work with microbial cultures. The phenomenon of being "out-of-phase" is described in the form of a newly defined nondimensional phase number (Ph) in analogy to a partially filled, rotating horizontal drum. The Ph can be used to determine reasonable operating conditions for shaking flask experiments when using viscous media, avoiding unfavorable "out-of-phase" operation. PMID- 10799985 TI - Application of metabolic flux analysis for the identification of metabolic bottlenecks in the biosynthesis of penicillin-G. AB - A detailed stoichiometric model was developed for growth and penicillin-G production in Penicillium chrysogenum. From an a priori metabolic flux analysis using this model it appeared that penicillin production requires significant changes in fluxes through the primary metabolic pathways. This is brought about by the biosynthesis of carbon precursors for the beta-lactan nucleus and an increased demand for NADPH, mainly for sulfate reduction. As a result, significant changes in flux partitioning occur around four principal nodes in primary metabolism. These are located at: (1) glucose-6-phosphate; (2) 3 phosphoglycerate; (3) mitochondrial pyruvate; and (4) mitochondrial isocitrate. These nodes should be regarded as potential bottlenecks for increased productivity. The flexibility of these principal nodes was investigated by experimental manipulation of the fluxes through the central metabolic pathways using a high-producing strain of P. chrysogenum. Metabolic fluxes were manipulated through growth of the cells on different substrates in carbon-limited chemostat culture. Metabolic flux analysis, based on measured input and output fluxes, was used to calculate the fluxes around the principal nodes. It was found that, for growth on glucose, ethanol, and acetate, the flux partitioning around these nodes differed significantly. However, this had hardly any effect on penicillin productivity, showing that primary carbon metabolism is not likely to contain potential bottlenecks. Further experiments were performed to manipulate the total metabolic demand for the cofactor nicotinamide adenine dinucleotide phosphate (NADPH). NADPH demand was increased stepwise by cultivating the cells on glucose or xylose as the carbon source combined with either ammonia or nitrate as the nitrogen source, which resulted in a stepwise decrease of penicillin production. This clearly shows that, in penicillin fermentation, possible limitations in primary metabolism reside in the supply/regeneration of cofactors (NADPH) rather than in the supply of carbon precursors. PMID- 10799986 TI - Two-phase model of the kinetics of growth of Rhizopus oligosporus in membrane culture. AB - An empirical model was developed to describe a growth profile occurring in solid state fermentation (SSF), namely that consisting of an initial period of rapid acceleration followed by an extended period of deceleration. This kinetic profile is not adequately described by the logistic model. The empirical model is based on the concept of active and nonactive hyphal segments. Exponential and deceleration growth phases are modeled. The model parameters can be determined directly from the dry-weight profile and they depend on the growth medium present. The model suggests that, at the instant the culture enters the deceleration phase, there is a 71% to 86% decrease in the number of actively extending hyphal tips and that, during the deceleration phase, there is an exponential decay in the number of active hyphal segments, with a first-order decay constant of 0.042 to 0.072 h(-1). PMID- 10799987 TI - Novel experimental study of receptor-mediated bacterial adhesion under the influence of fluid shear. AB - Dynamic adhesion of cells to surfaces is a vital step in a variety of biochemical and physiological phenomena. Bacterial adhesion is responsible not only for problems associated with biofouling and biofilm formation in the biochemical industry but also in the initiation of certain infectious diseases. In this study, we report the effect of critical parameters, such as receptor and ligand densities and shear rate, on receptor-mediated dynamic bacterial adhesion. Adhesion of a pathogenic strain of Staphylococcus aureus to immobilized collagen was studied. The receptor density on the cell surface was varied by harvesting cells at different growth times and was quantified using flow cytometry. Dynamic adhesion experiments were conducted over a range of physiologically relevant shear rates (50 to 1500 s(-1)) using a parallel-plate flow chamber. Video microscopy coupled with digital image processing was employed to quantify adhesion. A semiquantitative comparison between experimental results and theoretical data obtained using a previously proposed mathematical model was also performed. The results suggest that dynamic adhesion is dependent on receptor density and shear rate, but independent of ligand density. This report demonstrates the feasibility of using bacteria to study fundamental aspects of receptor-mediated dynamic adhesion. PMID- 10799988 TI - Ammonium alters N-glycan structures of recombinant TNFR-IgG: degradative versus biosynthetic mechanisms. AB - The effect of ammonium on the glycosylation pattern of the recombinant immunoadhesin tumor necrosis factor-IgG (TNFR-IgG) produced by Chinese hamster ovary cells is elucidated in this study. TNFR-IgG is a chimeric IgG fusion protein bearing one N-linked glycosylation site in the Fc region and three complex-type N-glycans in the TNF-receptor portion of each monomer. The ammonium concentration of batch suspension cultures was adjusted with glutamine and/or NH(4)Cl. The amount of galactose (Gal) and N-acetylneuraminic acid (NANA) residues on TNFR-IgG correlated in a dose-dependent manner with the ammonium concentration under which the N-linked oligosaccharides were synthesized. As ammonium increased from 1 to 15 mM, a concomitant decrease of up to 40% was observed in terminal galactosylation and sialylation of the molecule. Cell culture supernatants contained measurable beta-galactosidase and sialidase activity, which increased throughout the culture. The beta-galactosidase, but not the sialidase, level was proportional to the ammonium concentration. No loss of N glycans was observed in incubation studies using beta-galactosidase and sialidase containing cell culture supernatants, suggesting that the ammonium effect was biosynthetic and not degradative. Several biosynthetic mechanisms were investigated. Ammonium (a weak base) is known to affect the pH of acidic intracellular compartments (e.g., trans-Golgi) as well as intracellular nucleotide sugar pools (increases UDP-N-acetylglucosamine and UDP-N acetylgalactosamine). Ammonium might also affect the expression rates of beta1, 4 galactosyltransferase (beta1,4-GT) and alpha2,3-sialyltransferase (alpha2,3-ST). To separate these mechanisms, experiments were designed using chloroquine (changes intracellular pH) and glucosamine (increases UDP-GNAc pool [sum of UDP GlcNAc and UDP-GalNAc]). The ammonium effect on TNFR-IgG oligosaccharide structures could be mimicked only by chloroquine, another weak base. No differences in N-glycosylation were found in the product synthesized in the presence of glucosamine. No differences in beta1, 4-galactosyltransferase (beta1,4-GT) and alpha2,3-sialyltransferase (alpha2,3-ST) messenger RNA (mRNA) and enzyme levels were observed in cells cultivated in the presence or absence of 13 mM NH(4)Cl. pH titration of endogenous CHO alpha2,3-ST and beta-1,4-GT revealed a sharp optimum at pH 6.5, the reported trans-Golgi pH. Thus, at pH 7.0 to 7.2, a likely trans-Golgi pH range in the presence of 10 to 15 mM ammonium, activities for both enzymes are reduced to 50% to 60%. Consequently, ammonium seems to alter the carbohydrate biosynthesis of TNFR-IgG by a pH-mediated effect on glycosyltransferase activity. PMID- 10799989 TI - Microaerobic hydrogen production by photosynthetic bacteria in a double-phase photobioreactor. AB - The rate of hydrogen production by the marine nonsulfur photosynthetic bacterium, Rhodovulum sp., increased with increasing light intensity. A light intensity of 1800 W/m(2) hydrogen production rate was achieved at the rate of 9.4 micromol/mg dry weight/h. The hydrogen production of this strain was enhanced by the addition of a small amount of oxygen (12 micromol O(2)/reactor). Intracellular ATP content was most efficiently accumulated under microaerobic, dark conditions. Hydrogen production rate by Rhodovulum sp. was investigated using a double-phase photobioreactor consisting of light and dark compartments. This rate was compared with data obtained using a conventional photobioreactor. Rhodovulum sp. produced hydrogen at a rate of 0.38+/-0.03 micromol/mg dry weight/h under microaerobic conditions using the double-phase photobioreactor. The hydrogen production rate was four times greater under microaerobic conditions, as compared with anaerobic conditions using either type of photobioreactor. Hydrogen production using a double-phase photobioreactor was demonstrated continuously at the same rate for 150 h. PMID- 10799990 TI - Metabolic network analysis of Penicillium chrysogenum using (13)C-labeled glucose. AB - Using (13)C-labeled glucose fed to a penicillin-overproducing strain of Penicillium chrysogenum, the intracellular fluxes were quantified, and the presence of two new pathways, not previously described in this organism, is suggested. Thus, glycine was synthesized not only by serine hydroxymethyltransferase, but also by threonine aldolase. The formation of cytosolic acetyl-CoA was found to be synthesized both via the citrate lyase catalyzed reaction and by degradation of the penicillin side-chain precursor, phenoxyacetic acid. Furthermore, the experimental data indicate that the main activities of homocitrate synthase and alpha-isopropylmalate synthase are located in the cytosol. All experimental data on the labeling patterns were obtained using gas chromatography-mass spectrometry, which is faster and more sensitive than the nuclear magnetic resonance methods usually applied for analysis of labeling patterns. PMID- 10799991 TI - Hydrolysis of glucosinolates using nylon-immobilized myrosinase to produce pure bioactive molecules. AB - Bioactive compounds were produced from natural glucosinolates, secondary plant metabolites, using myrosinase (thioglucoside glucohydrolase EC 3.2.3.1) isolated from ripe seeds of Sinapis alba. The enzyme was immobilized on granular nylon 6.6 with the crosslinking technique. Immobilized myrosinase displayed extraordinary operational and storage stability. Using a small thermostatted continuous packed bed bioreactor, the enzyme activity was unchanged after 15 days of continuous use at 37 degrees C and after >1 year of storage at room temperature. The bioreactor was particularly efficient in producing pure isothiocyanates, but it was less efficient for pure nitrile production. PMID- 10799992 TI - Investigating the effects of polymer chemistry on activity of biocatalytic plastic materials. AB - The affects of polymer chemistry on the organic solvent activity of alpha chymotrypsin-containing biocatalytic plastic materials are investigated in this study. To incorporate alpha-chymotrypsin into the polymer, the enzyme is first acryloylated, then solubilized into organic solvents via hydrophobic ion paring with surfactant molecules. Once in the organic solvent, a vinyl monomer and crosslinker are added and copolymerized with the enzyme. Due to the intimate contact between the enzyme and the resulting polymer network, the polymer chemistry plays an important role in the activity of these biocatalytic materials. The chemical composition of the monomer/polymer has the greatest effect on catalytic activity. The activity spans a range of 100-fold and appears to correlate with the hydrophilicity of the monomer, with the lowest activity exhibited for poly(methyl methacrylate) and the highest for poly(2-hydroxyethyl methacrylate). The effect of the chemical structure of the monomer/polymer appears to be an intrinsic kinetic effect, whereas other polymer chemistry conditions investigated, including crosslinker concentration and length and ratio of solvent:monomer during synthesis, appear to effect the rate of substrate diffusion, thereby affecting observed enzyme activity. Changes in the conditions of polymer synthesis can cause as much as a 20-fold change in activity for a given polymeric material. This is most likely due to an increase in the porosity of the materials, and thus a relaxation of diffusional limitations. PMID- 10799993 TI - Hydrophobic displacement chromatography of proteins. AB - Displacement chromatography of proteins was successfully carried out in both hydrophobic interaction and reversed-phase chromatographic systems using low molecular weight displacers. The displacers employed for hydrophobic displacement chromatography were water soluble, charged molecules containing several short alkyl and/or aryl groups. Spectroscopy was employed to verify the absence of structural changes to the proteins displaced on these hydrophobic supports. Displacement chromatography on a reversed-phase material was employed to purify a growth factor protein from its closely related variants, demonstrating the high resolutions that can be achieved by hydrophobic displacement chromatography. This process combines the high-resolution/high-throughput characteristics of displacement chromatography with the unique selectivity of these hydrophobic supports and offers the chromatographic engineer a powerful tool for the preparative purification of proteins. PMID- 10799994 TI - A vortex-bowl disk atomizer system for the production of alginate beads in a 1500 liter fermentor. AB - Using a model system, a concept for the immobilization of microbial cultures within alginate beads directly in a 1500-L fermentor with a height to diameter ratio of 1.85 is described. The system is comprised of a 60-cm diameter bowl fixed to the top of an agitation shaft, where calcium-ion-rich media is continuously recirculated from the bulk solution to the bowl. The rotation of the shaft and bowl creates a climbing film (vortex) of solution. An atomizing disk centrally recessed within the bowl sprays an alginate solution into the climbing film where the droplets harden into beads. The effect of heat treatment on the alginate solution on resulting bead properties was examined. The sterilization operation did not appear to have a major effect on the alginate bead mechanical properties of firmness and elasticity which was much more a function of alginate concentration. Beads of various sizes were produced by the unit. The system was characterized by the dimensionless numbers Reomega = (omega x rho x D(2))/mu and ReQ = (Q x rho)/(mu x D). At Reomega and ReQ values less than 500 and 0.15, respectively, the mechanism was direct drop. Parent droplets followed by satellite droplets were observed. When either the flow rate or speed was increased, filaments formed predominantly, which was unwanted in this system because filament breakdown into smaller droplets does not occur due to the proximity of the disk to the climbing film in the bowl. This system could be applied to the immobilization of microorganisms, as well as plant or animal cell cultures, and for other sizes or fermentors. The overflow from the bowl carries the gellified beads into the bulk solution where immobilized cells could act upon the fermentation media. PMID- 10799995 TI - Exploiting viral cell-targeting abilities in a single polypeptide, non infectious, recombinant vehicle for integrin-mediated DNA delivery and gene expression. AB - A recombinant, multifunctional protein has been designed for optimized, cell targeted DNA delivery and gene expression in mammalian cells. This hybrid construct comprises a viral peptide ligand for integrin alpha(V)beta(3) binding, a DNA-condensing poly-L-lysine domain, and a complete, functional beta galactosidase protein that serves simultaneously as purification tag and DNA shielding agent. This recombinant protein is stable; it has been produced successfully in Escherichia coli and can be purified in a single step by affinity chromatography. At optimal molar ratios, mixtures of this vector and a luciferase reporter plasmid form stable complexes that transfect cultured cells. After exposure to these cell-targeted complexes, steady levels of gene expression are observed for more than 3 days after transfection, representing between 20 and 40% of those achieved with untargeted, lipid-based DNA-condensing agents. The principle to include viral motifs for cell infection in single polypeptide recombinant proteins represents a promising approach towards the design of non viral modular DNA transfer vectors that conserve the cell-target- ing specificity of native viruses and that do not need further processing after bioproduction in a recombinant host. PMID- 10799996 TI - Observations on the shear damage to different animal cells in a concentric cylinder viscometer. AB - A clear distinction is made between damage to the population of cells and damage to individual cells on exposure to shear stress. Work on mechanical damage to animal cells in suspension is reported for six different cell lines. Precisely controlled shears of 1 Pa, 10 Pa, and 100 Pa were generated in a viscometer and distortions in morphology of the cells-for instance, the formation of transient pores, cytoplasmic extrusions, and ghost-cell membranes-are presented with photographic evidence. Low shears are shown to be just as damaging as the higher shears, although the type of damage is different. It follows that bioreactors should be operated at intermediate shear levels for optimal yield. A mechanism to account for the unexpected stability of animal cells at intermediate levels of shear is presented. PMID- 10799997 TI - Mathematical description of microbiological reactions involving intermediates PMID- 10799998 TI - Sonography of palpable breast cancer. AB - PURPOSE: Because of the increasing use of sonography to rule out cancer in women with palpable breast abnormalities, this study was performed to determine the rate of sonographically occult malignancy in this clinical setting. METHODS: Women who were recommended for biopsy based on mammographic and/or clinical findings underwent breast sonography. This study retrospectively analyzed the subset of patients with palpable malignant lesions. Lesions were classified as visible or occult on mammography and sonography. Patients without a tissue diagnosis of tumor were excluded. RESULTS: Of 1,346 masses that underwent biopsy or aspiration, 616 lesions were palpable, and of these, 293 were malignant. Sonography detected all 293 palpable malignant lesions (95% confidence interval for sensitivity, 99-100%). Eighteen lesions were mammographically occult. The median lesion size as determined by sonography was 1.8 cm; for the lesions that were mammographically occult, the median size was 1.6 cm. The most common histopathologic diagnosis for both groups of lesions was infiltrating ductal carcinoma. CONCLUSIONS: All palpable malignant breast lesions were visible by sonography in patients in whom a biopsy was recommended. However, we caution that until the false-negative rate of sonography for equivocal palpable abnormalities is determined prospectively, sonography cannot be accurately applied to rule out malignancy in this setting. PMID- 10799999 TI - Comparison of transvaginal sonography, saline infusion sonography, and hysteroscopy in premenopausal women with abnormal uterine bleeding. AB - PURPOSE: Saline infusion sonography (SIS) is a relatively new technique in the evaluation of abnormal uterine bleeding. We compared the diagnostic accuracy of SIS with that of transvaginal sonography (TVS) in the detection of intracavitary abnormalities in premenopausal women with abnormal uterine bleeding. METHODS: In this prospective study, consecutive premenopausal women who underwent a hysteroscopy for abnormal uterine bleeding also underwent TVS and SIS. The findings at TVS and SIS were compared with the hysteroscopic and histologic findings. Sensitivity, specificity, and likelihood ratios were calculated. Receiver operating characteristic curves were constructed to assess the performance of endometrial thickness measured using TVS. RESULTS: Sixty-two patients were included in the study. TVS demonstrated 60% sensitivity in directly visualizing intracavitary abnormalities and 93% specificity. The likelihood ratio of the presence of an intracavitary abnormality was 8, and the likelihood ratio of the absence of an intracavitary abnormality was 0.43. Defining an abnormality at TVS as direct visualization of an intracavitary abnormality or an endometrial thickness greater than 5 mm, TVS had an 85% sensitivity and a 21% specificity, with corresponding likelihood ratios of 1.1 and 0.71, respectively. For SIS, the sensitivity, specificity, and likelihood ratios of the presence and absence of intracavitary abnormalities were 88%, 95%, 10, and 0.13, respectively. CONCLUSIONS: SIS is more accurate in the diagnosis of intracavitary abnormalities in premenopausal women than is TVS. An approach using endometrial thickness measurement by TVS and reserving SIS for patients who have an endometrial thickness greater than 5 mm or an intracavitary abnormality visualized by TVS would be the most effective method to reduce the number of hysteroscopies. PMID- 10800000 TI - Effect of antenatal steroid administration on the fetal biophysical profile. AB - PURPOSE: Our objective was to determine whether antenatal steroid administration affects the biophysical profile score in fetuses. METHODS: A prospective study was conducted in 84 fetuses between 28 and 34 weeks' menstrual age at risk of preterm delivery. Two intramuscular injections of 12 mg of betamethasone were given to the mother 24 hours apart. All fetuses underwent biophysical profile testing prior to and between 24 and 48 hours after steroid administration. Biophysical profiles (including nonstress tests) were evaluated by two maternal fetal medicine specialists blinded to the timing of steroid administration. Neonatal outcome, including Apgar score, menstrual age at delivery, admission to and length of stay in the neonatal intensive care unit, and mortality, was analyzed in all subjects. RESULTS: In 31 (37%; 95 confidence interval, 26.6 47.2%) of 84 cases, the biophysical profile score decreased at least 2 points after steroid administration. The most commonly affected variables were fetal breathing and the nonstress test. There was no significant difference in the neonatal outcome between the fetuses whose biophysical profile decreased and those whose did not. CONCLUSIONS: Biophysical profile scores were decreased in more than one third of fetuses within 48 hours of antenatal steroid administration, but neonatal outcome was not affected. Knowledge of this occurrence could avoid incorrect decision making regarding fetal well-being. PMID- 10800001 TI - Carcinoma of the gallbladder: role of sonography in diagnosis and staging. AB - PURPOSE: In an attempt to define the sonographic characteristics of gallbladder cancer, we retrospectively analyzed the sonographic findings in 203 cases of gallbladder cancer confirmed by cytology or histopathology. Patients and Methods Patients with proven gallbladder cancer presenting to a single surgical unit between 1991 and 1995 were identified through a records search. All patients underwent sonographic examination followed by fine-needle aspiration (FNA), biopsy, or laparotomy for establishing the diagnosis. RESULTS: A mass in the gallbladder and gallbladder wall thickening (> 12 mm) were cardinal sonographic findings of carcinoma. Liver infiltration was correctly identified in all patients who had it. Sonography was highly accurate for detecting mass lesions, gallstones, liver infiltration, metastasis, and ascites. However, visualization of lymph nodes, common bile duct infiltration, and peritoneal dissemination was poor. CONCLUSIONS: Sonography was found to be a good diagnostic tool for carcinoma of the gallbladder; however, its sensitivity was poor for staging nodal spread of the disease. PMID- 10800002 TI - Sonographic appearances in abdominal tuberculosis. PMID- 10800003 TI - Power Doppler sonographic diagnosis of torsion in a wandering spleen. AB - A wandering spleen is a rare clinical entity resulting from congenital maldevelopment or acquired laxity of the spleen's suspensory ligaments; it may result in torsion of the spleen. We report the gray-scale sonographic, power Doppler sonographic, and CT findings in a case of wandering spleen torsion. The gray-scale sonograms showed a displaced spleen that appeared as a homogeneous, hypoechoic mass suggestive of an enlarged, ectopic spleen in the central abdomen. Power Doppler sonograms showed no blood flow in the parenchyma or hilum of the spleen and were consistent with torsion and infarction in the spleen. Contrast enhanced CT scans showed a homogeneous, unenhanced mass that was diagnosed as a torsioned wandering spleen. The hilar vessels of the spleen were also unenhanced. PMID- 10800004 TI - Color Doppler sonographic findings in splenic hamartoma. AB - We present the gray-scale and color Doppler sonographic findings in a case of a splenic hamartoma in a 40-year-old man. Gray-scale sonograms showed a 2 x 2 cm, hypoechoic splenic mass that was homogeneous without evidence of cystic change or calcification. Color Doppler sonograms showed multiple radial blood-flow signals inside the mass, and spectral analysis confirmed arterial and venous flow. Arteriograms showed multiple small, hypervascular masses with fine tumor vessels and tumor stains within the spleen. Histologic analysis following a splenectomy showed dilated vessels and congestion consistent with the color Doppler sonographic findings. PMID- 10800005 TI - Sonographic appearance of the Angelchik prosthesis. AB - The Angelchik prosthesis is a silicone-filled, doughnut-shaped device used for the surgical treatment of gastroesophageal reflux. This device appears on sonograms as a C-shaped, hypoechoic structure with nonspecific Doppler waveforms. Because the Angelchik prosthesis is rarely used anymore, recently trained sonographers and radiologists may misinterpret this sonolucent appearance as an unusual vascular lesion. We report a case of an Angelchik prosthesis that was incidentally noted at the gastroesophageal junction on a routine abdominal sonographic examination. PMID- 10800006 TI - Hepatic hemangioendothelioma: prenatal sonographic findings and evolution of the lesion. AB - We describe a case of hepatic hemangioendothelioma that was first suspected based on prenatal sonographic findings at 19 weeks' menstrual age. At 16 weeks, the patient presented with a markedly elevated maternal serum alpha-fetoprotein level. Serial sonographic examinations revealed that the fetus had cardiomegaly, hepatomegaly with a hepatic mass and dilated intrahepatic vessels, a single umbilical artery, and a placental chorioangioma. Arteriovenous shunting within the hepatic mass was seen using color Doppler and pulsed Doppler sonography. An enlarged artery arising from the abdominal aorta supplying the mass was demonstrated. Postnatal physical examination and radiologic studies supported the diagnosis of hepatic hemangioendothelioma. The evolution in the sonographic appearance of this hepatic lesion in utero over a 17-week period is described. PMID- 10800007 TI - Subdiaphragmatic esophageal duplication cyst in a child. PMID- 10800008 TI - Establishing guidelines for pharmacotherapy trials in bulimia nervosa and anorexia nervosa. AB - OBJECTIVE: This paper addresses the lack of a standard protocol for pharmacotherapy trials for patients with bulimia nervosa (BN) and anorexia nervosa (AN). METHOD: Twenty-two surveys were sent to established researchers in the field of eating disorders to elicit their opinions regarding medication trials, including baseline laboratory tests, the optimal length/frequency of medication management sessions, and the information that should or should not be included in these sessions. RESULTS: Sixteen of 22 researchers completed and returned the survey. Their answers are the basis of the data presented. DISCUSSION: We propose a battery of screening laboratory tests for both conditions. We suggest 30-45-min initial medication management sessions in both AN and BN trials with 15-min follow-ups to be held weekly for AN subjects, and weekly for 2 weeks, then biweekly for 2 weeks, then monthly, for BN subjects. We also recommend that published trials should include explicit details of medication management. PMID- 10800009 TI - Inhibited expression of negative emotions and interpersonal orientation in anorexia nervosa. AB - OBJECTIVE: This study examined inhibited expression of negative feelings and interpersonal orientation in women with anorexia nervosa. METHOD: Twenty-one women meeting DSM-IV criteria for anorexia nervosa were compared with 21 psychiatric and 21 normal control women matched on education. Two measures were used to assess inhibited expression of negative feelings and interpersonal orientation: the State-Trait Anger Expression Inventory assesses the suppression and expression of anger and the Silencing the Self Scale assesses four cognitive schemas involving the repression of needs and feelings to protect interpersonal relationships. RESULTS: Women with anorexia nervosa reported significantly higher scores on the four Silencing the Self schemas and on suppressed anger after controlling for age. These group differences were maintained for two of the cognitive schemas (Care and Silence) after controlling for depression, self esteem, and global assessment of functioning. Inhibited expression of negative emotion and interpersonal orientation scores were also significantly related to cognitive and affective components of body image dissatisfaction and to trait and self-presentational dimensions of perfectionism. DISCUSSION: These findings are reviewed in the context of health psychology, as well as feminist and temperament theories. Implications for treatment are addressed. PMID- 10800010 TI - Mortality in eating disorders: a descriptive study. AB - OBJECTIVE: We report rates and causes of death for a cohort of 246 eating disordered women and provide descriptive information on their eating disorder and comorbid diagnoses. METHOD: Data on mortality were collected as part of a longitudinal study of anorexia nervosa and bulimia nervosa, now in its 11th year. Other data sources included death certificates, autopsy reports, relative interviews, and a National Death Index search. RESULTS: Seven deaths have occurred during the study, all among anorexic subjects with a history of binging and purging and with comorbid Axis I disorders. The crude mortality rate was 5.1%. The standardized mortality ratios for death (9.6) and suicide (58.1) were significantly elevated (p <. 001). CONCLUSIONS: Anorexia nervosa is associated with a substantial risk of death and suicide. Features correlated with fatal outcome are longer duration of illness, binging and purging, comorbid substance abuse, and comorbid affective disorders. PMID- 10800011 TI - Developmental onset of eating-related color-naming interference: the role of restraint and eating psychopathology. AB - OBJECTIVE: This study investigated the developmental onset of the Stroop interference effect for food and body shape words in 12- and 14-year-old females to determine whether dietary restraint and eating psychopathology influenced Stroop performance times. METHOD: A Stroop task containing neutral, food, and body shape-related words was administered to 152 schoolgirls. Participants completed the restraint scale of the Dutch Eating Behavior Questionnaire and the Drive for Thinness (DFT) subscale of the Eating Disorders Inventory. RESULTS: Significant color-naming impairments were observed for food-related words in 12- and 14-year-olds, in 14-year-old restrained eaters, and in 12-year-old unrestrained eaters. There were no significant differences between restrained and unrestrained eaters in either age group. Participants scoring high on the DFT subscale showed significant impairments for food-related words, but did not differ significantly from those scoring low on the DFT. There were no significant impairments in color-naming body shape-related words in any subgroups. DISCUSSION: This study did not confirm a different developmental onset for the food and body shape Stroop interference effect. Consideration of dietary restraint did add clarity to previously observed food-related interference effects in both age groups. The results for high- and low-DFT participants do not support the use of the eating-related Stroop tests as an early objective indicator of eating psychopathology. PMID- 10800013 TI - Improving the body image, eating attitudes, and behaviors of young male and female adolescents: a new educational approach that focuses on self-esteem. AB - OBJECTIVE: This study examines the effect of an interactive, school-based, self esteem education program on the body image and eating attitudes and behaviors of young male and female adolescents following the program and after 12 months. METHOD: All 470 eligible students (63% female) aged 11-14 years volunteered to participate. The intervention group students participated in the program, whereas the control group students received their scheduled personal development and health class. RESULTS: The program significantly improved the body satisfaction of the intervention students and significantly changed aspects of their self esteem; social acceptance, physical appearance, and athletic ability became less important for the intervention students and more important for control students. Female intervention students rated their physical appearance as perceived by others significantly higher than control students and allowed their body weight to increase appropriately by preventing the age increase in weight-losing behaviors of the control students. One year after the intervention, body image and attitude changes were still present. These findings also held for the 116 students (63% females) with low self-esteem and higher anxiety, who were considered at risk for the development of eating disorders. These students also had significantly lower drive for thinness and greater body satisfaction following the intervention and the decreased importance of physical appearance to their self-esteem was present at 12 months. Control at-risk students significantly decreased their body weight, whereas the weight of the intervention at-risk students significantly increased. The intervention program was effective, safe, having no effect on measures of students' anxiety or depression, and was rated highly by students. DISCUSSION: This is the first controlled educational intervention to successfully improve body image and to produce long-term changes in the attitudes and self-image of young adolescents. This new approach to prevent the development of eating disorders by improving self-esteem may be effective, particularly if reinforced by teachers and family. PMID- 10800012 TI - Psychological and behavioral factors unpredictive of disordered eating: a prospective study of the general adolescent population in Norway. AB - OBJECTIVE: To investigate if cross-sectionally identified correlates of disordered eating among adolescents could also predict disordered eating prospectively over and beyond the initial level of disordered eating. METHOD: Two wave longitudinal questionnaire study of a representative and nationwide sample of 7,751 Norwegian adolescents aged 12-19 at initial testing (t1). A 12-item version of the Eating Attitudes Test (EAT) was included. RESULTS: Forty percent of the girls and 25% of the boys with disordered eating at t1 also scored above the cut-off point 2 years later (t2). Initial disordered eating, gender, depressed mood, body dissatisfaction, unstable self-perceptions, perceived obesity, excessive exercise, and having idols with perfect bodies all predicted change in disordered eating. However, when the initial symptom load was controlled, these variables - except gender - only contributed marginally to the prediction of disordered eating. These negative results are in line with other longitudinal studies of changes in disordered eating in unselected adolescent populations. DISCUSSION: It cannot be established that psychological factors play a major etiological role in the development of disordered eating. The results are discussed with reference to a potential discontinuity between eating problems and eating disorders. PMID- 10800014 TI - Psychiatric comorbidity associated with eating disorder symptomatology among adolescents in the community. AB - OBJECTIVE: This study investigates psychiatric comorbidity associated with eating disorder symptomatology among adolescents in the community. METHOD: Four hundred three adolescents in the community were administered structured clinical interviews to assess mood, anxiety, eating, substance use, and personality disorders. RESULTS: Adolescents with dysthymia, panic and major depressive disorder were significantly more likely than those without these disorders to have an eating disorder. After controlling for the effects of other Axis I disorders and personality disorders, only dysthymia independently predicted the presence of an eating disorder. Several personality disorders were also associated with eating disorder symptoms. However, only obsessive-compulsive personality disorder predicted eating disorder symptoms after controlling for other personality disorders. CONCLUSION: Although previous research on adults has focused on the association between major depressive disorder and eating disorders, dysthymia may be more strongly associated with eating disorders among adolescents in the community. This association is not accounted for by psychiatric comorbidity. PMID- 10800015 TI - Relationship of weight and eating disorders in type 2 diabetic patients: a multicenter study. AB - OBJECTIVE: Previous work suggested that the degree of psychiatric symptomatology evidenced in overweight individuals was related to the severity of binge eating problems and not related to the severity of overweight. In a multicenter study, we investigated the relationship between weight and eating disorders (EDs) in a sample of type 2 diabetic patients. METHODS: Three hundred twenty-two patients with type 2 diabetes were stratified to various weight categories. Glycemic control, eating and body-related psychological problems, self-esteem, depressive, and general psychopathology of diaetic patients with and without an ED were compared. RESULTS: Eighty-one percent of all type 2 diabetic patients were overweight or obese. Prevalence rates of EDs ranged from 6.5% to 9.0%. Binge eating disorder was the most diagnosed ED. There was a strong relationship between body mass index (BMI) and eating disturbance-related variables and a weak or no relationship between BMI and depression or general psychopathologic variables. Patients with an ED showed a greater psychopathology compared to patients without an ED. The diagnosis of an ED did not seem to have a specific influence on glycemic control. CONCLUSIONS: Our results in a type 2 diabetic sample indicate that weight might have an impact on body and eating-related psychological distress in type 2 diabetic patients, but is of minor or no importance for depressive symptomatology, lower self-esteem, and general psychiatric symptomatology. Type 2 diabetic patients with an ED, however, suffer from considerable psychiatric symptomatology. PMID- 10800016 TI - The role of the mother-daughter relationship in explaining weight concern. AB - OBJECTIVE: The literature highlights two possible roles for the mother-daughter relationship in the development of weight concern in young girls. The first is simply as a forum for modeling the mother's own concerns and the second is as an interaction between two individuals, which is either protective or facilitative of weight concern. The present study aimed to assess both the modeling and the interactive hypotheses. METHOD: Thirty mothers and their daughters (aged 16-19) completed a questionnaire concerning their weight concern (restrained eating and body dissatisfaction) and their beliefs about aspects of their relationship (daughter's autonomy, mother's autonomy, projection, intimacy, mother's role). RESULTS: The results found no support for the modeling hypothesis, with no significant correlations between the mothers' and daughters' aspects of weight concern. However, the results showed support for the interactive hypothesis. In particular, the daughters were more likely to show restrained eating if their mothers reported a low belief in their own autonomy and if both the mother and daughter rated projection as important in their relationship. Similarly, the daughters were more likely to show body dissatisfaction if their mothers reported a low belief in both their own and their daughter's autonomy and if they rated projection as important. CONCLUSION: The results indicate that the role of the mother-daughter relationship may be more complex than simply as a forum for modeling and are discussed in terms of the impact of aspects of the relationship on subsequent psychopathology. PMID- 10800017 TI - Family dysfunction and bulimic psychopathology: the mediating role of shame. AB - OBJECTIVE: Although disturbed family function has some association with bulimic psychopathology, the psychological mechanisms that account for that link are not clear. This study explores the hypothesis that shame acts as a mediator in that relationship, whereas shame-proneness is a moderator variable. METHOD: The participants were 139 nonclinical women. Each completed measures of perceived family function, shame-proneness, internalized shame, and bulimic psychopathology. Regression analyses were used to test for the mediating and moderating effects of shame. RESULTS: The findings were compatible with a model where shame-proneness acts as a moderator and internalized shame is a perfect mediator in the link between paternal overprotection and bulimic attitudes. CONCLUSIONS: The experience of shame appears to be a critical element in understanding the relationship between perceived family dysfunction and bulimic psychopathology. Where individuals perceive their families as problematic, it may be clinically valuable to focus on shame as a psychological consequence of that experience. PMID- 10800018 TI - Continuity and change in the evaluation of ideal and acceptable body sizes across a wide age span. AB - OBJECTIVE: Continuity and change in the evaluation of ideal and acceptable body sizes across a wide subject age span were examined. METHOD: Ratings of ideal and socially acceptable body sizes were elicited from 303 children, 427 adolescents, 261 young adults, and 326 middle-age adults. Line drawing arrays of babies, children, young adults, middle-age, and older adults were portrayed, ranging in size from very thin to very obese. RESULTS: All subject groups selected, in all arrays, similar ideal body sizes, rated sizes in the midrange of fatness as socially acceptable, and were least accepting of very thin and obese body sizes. Tolerance for body size variations increased with subject age. DISCUSSION: Continuity throughout a wide subject age span was observed in evaluations of body sizes. However, adults were more accepting of body size variations than younger subjects, especially children. Implications of endorsing midrange body sizes for the fashion industry are discussed. PMID- 10800019 TI - Pilot study of a graded exercise program for the treatment of anorexia nervosa. AB - OBJECTIVE: To determine whether a graded exercise program used in the treatment of anorexia nervosa improves quality of life and does not decrease the rate of gain of body fat. METHODS: A randomized controlled trial with outcome measures: change in percent body fat, body mass index (BMI), and Medical Outcomes Survey Short Form 36-item Quality of Life questionnaire. RESULTS: Fifteen females and one male meeting the DSM-IV criteria for the diagnosis of anorexia nervosa were randomized. There was no difference in change in BMI or percent body fat at 3 months. Quality of life outcomes improved from baseline in the experimental group compared with the control group. However, this difference was not statistically significant. DISCUSSION: Incorporation of a graded exercise program may increase compliance with treatment, but it did not reduce the short-term rate of gain of body fat or BMI. Longer studies with more subjects are necessary to determine the usefulness of a graded exercise program in anorexia nervosa. PMID- 10800021 TI - Assessing the relevance of the hopelessness theory of depression to women with disordered eating. AB - OBJECTIVE: The aim of this study was to investigate the relevance of the hopelessness theory of depression to women with partial-syndrome eating disorders. METHOD: Three groups of women, one meeting criteria for eating disorders not otherwise specified (EDNOS), one with major depression, and a control group, completed the Balanced Attributional Style Questionnaire, the Eating Disorders Examination Questionnaire, and the Beck Depression Inventory. RESULTS: Once levels of depression were controlled, all three groups differed significantly with respect to their attributional style for bad events, with the depressed group showing the greatest tendency to attribute the causes of negative life events to internal factors. However, with respect to the attributional style for good events, the depressed and control group displayed similar styles of attribution, whereas the EDNOS group showed a significantly more dysfunctional style, being more likely to attribute positive events to external factors. DISCUSSION: These findings are discussed in terms of their implications for specific therapeutic interventions with disordered eating. PMID- 10800020 TI - Bone density of women who have recovered from anorexia nervosa. AB - OBJECTIVE: To examine bone density in 19 women who had previously experienced classical anorexia nervosa from which they had been fully recovered for a mean of 21 years (current characteristics: median age 40.2 years; Eating Attitudes Test [EAT] score 2; body mass index [BMI] 21.1; average 1.8 offspring). METHOD: Probands were compared, blindly, in respect of bone density, with 13 control subjects matched for age and sex and with no history of eating disorders. Dual energy X-ray absorptiometry (DXA) was used to evaluate the bone mineral density (BMD) of the lumbar spine and the head of the femur. RESULTS: Femur BMD was still significantly less among ex-anorectic sufferers. Two subjects had experienced pathological fractures while anorectic, both having been strenuous exercisers. None appeared to have suffered post illness fractures. BMD at follow-up did not relate to the severity or chronicity of previous anorexia nervosa. DISCUSSION: Full clinical recovery from anorexia nervosa does not quite confer full establishment of normal bone density. However, pathological fractures are not a feature thereafter, within middle life. PMID- 10800022 TI - Investigating muscularity concerns in male body image: development of the Swansea Muscularity Attitudes Questionnaire. AB - OBJECTIVE: An investigation into muscularity concerns in males was carried out. METHOD: A series of questionnaires, containing items pertaining to attitudes and behavior relating to muscularity, were administered to normal male subjects. RESULTS: Factor analysis revealed two underlying constructs. The first reflected the desire to be muscular and and to such goal-oriented behaviors. The second reflected perceived positive attributes of muscularity, including feelings of masculinity, enhanced confidence, and greater attractiveness. Following preliminary studies, a final version of the Swansea Muscularity Attitudes Questionnaire was developed and administered to 303 normal male subjects. Factor analysis of these data confirmed the dual-construct model. The two subscales of this questionnaire (Drive for Muscularity and Positive Attributes of Muscularity) displayed high internal reliability. DISCUSSION: It is suggested that this questionnaire provides a tool that may be used to investigate further the relationship between muscularity concerns and body image issues in males. PMID- 10800023 TI - Acculturation and disordered eating patterns among Mexican American women. AB - OBJECTIVE: This quantitative study examined the relationship between acculturation and disordered eating patterns among different generations of Mexican American women. METHOD: Participants included 139 Mexican American women (mean age = 29.1) drawn from local undergraduate courses and community agencies. The posttest-only design included a demographic measure, Acculturation Rating Scale for Mexican Americans (ARSMA), and the Eating Attitudes Test (EAT-26). RESULTS: Of the five generations studied, second-generation women endorsed the most disordered eating patterns and the highest degrees of concurrent acculturation. Moreover, there was a significant, positive correlation between acculturation and Factor III of the EAT which highlights control of eating and perceived pressure from others to gain weight. Participants scoring beyond the EAT cut-off score endorsed bulimic symptoms. DISCUSSION: Differing intergenerational ideals regarding weight may create special strains for second generation Mexican American women. Criteria for disordered eating in a population previously not considered at risk should be further assessed and clinicians should be careful not to underdiagnose among this group. PMID- 10800024 TI - In vivo glucose detection by homonuclear spectral editing. AB - A frequency-selective multiple-quantum-coherence spectral editing pulse sequence, Ssel-MQC, was implemented for the detection of the betaH1-glucose resonance at 4.63 ppm in rat brain in vivo. Unwanted signal suppression and glucose coherence transfer pathway selection were performed with magnetic field gradients. To optimize sensitivity, the sequence was executed with surface coil signal reception and adiabatic RF pulse transmission. The glucose editing capabilities of Ssel-MQC were first evaluated in vitro. Ssel-MQC achieved excellent water suppression (suppression factor >10(5)), at the expense of an approximately 60% loss of the glucose signal due to incomplete coherence transfer pathway selection. Next, the sequence was used for in vivo glucose detection in normal rat brain during D-glucose infusion and in the brain of diabetic rats prior to and following insulin infusion. PMID- 10800025 TI - Intermolecular zero-quantum coherence imaging of the human brain. AB - The first intermolecular zero-quantum coherence (iZQC) MR images of the human brain at 4T are presented. To generate iZQC images, a modified echo-planar imaging pulse sequence was used which included an additional 45 degrees RF pulse and a correlation gradient. The observability and nonconventional contrast of human brain iZQC images at 4T is demonstrated. Axial images are presented for various pulse sequence parameters, and a zero-quantum relaxation map is obtained. PMID- 10800027 TI - In vivo (1)H double quantum filtered MRI of the human wrist and ankle. AB - Proton double quantum filtered (DQF) NMR imaging was applied in vivo to the human wrist and ankle with a clinical 1.5 T MR scanner. Water molecules having anisotropic motion were detected from tendons and ligaments. Images of Achilles tendon were obtained for a voxel size of 1.25 x 1.25 x 20 mm with three values of TR = 1.0, 0.5, and 0.2 sec, resulting in total acquisitions time of 17, 8.5, and 3.4 mins, respectively. Some degradation of the signal-to-noise ratio was observed at the shortest TR value and the contrast was significantly reduced due to SQ coherence leakage. The in vivo DQF images showed structure within the tendon that is otherwise not visible by conventional gradient-recalled echo (GRE) methods. Published 2000 Wiley-Liss, Inc. PMID- 10800026 TI - Uptake of a superparamagnetic contrast agent imaged by MR with high spectral and spatial resolution. AB - Conventional MRI implicitly treats the proton signal as a single, narrow Lorentzian. However, water signals in vivo are often in homogeneously broadened and have multiple resolvable components. These components represent discrete populations of water molecules within each pixel which are affected differently by physiology and contrast agents. Accurate measurement of each component of the water resonance can improve anatomic and functional MR images and provide insight into the structure and dynamics of subpixelar microenvironments. This report describes high spectral and spatial resolution (HiSS) MR imaging of rodent prostate tumors before and after injection of a superparamagnetic contrast agent. HiSS datasets were used to synthesize images in which intensity is proportional to peak height, peak frequency, and linewidth. These images showed anatomic features which were not clearly delineated in conventional T(2) and gradient echo images. HiSS images obtained after injection of the contrast agent showed T *(2) and T(1) changes which were not seen in conventional images. These changes are associated with microvessel density and permeability. The results suggest HiSS with superparamagnetic contrast agents has the potential to improve characterization of tumors. PMID- 10800028 TI - TE-switched double-contrast enhanced visualization of vascular system and instruments for MR-guided interventions. AB - A visualization principle for MR-guided vascular interventions based on the concerted use of two contrast agents is introduced. The first contrast agent, consisting of small paramagnetic iron oxide particles, was administered intravenously to shorten T(1), and even more so T *(2), of the blood for extended time periods. The second agent, a monomeric gadolinium complex, was added to a solution in an interventional device, such as a percutaneous-transluminal angioplasty (PTA) balloon, to reduce T(1) with only minor additional effects on T *(2). With appropriate T(1)- and T *(2)-weighting the vascular tree (TE <3 ms) and the device (TE > or =8 ms) could be selectively imaged (TR <20 ms). Potentially, both images could be simultaneously updated in the subsecond range. Whereas a visualization of thin structures like guidewires was found to require pulse-sequence optimization, a successful visualization of a PTA balloon in a swine aorta in vivo was possible with standard sequences. PMID- 10800029 TI - Lactate editing and lipid suppression by continuous wavelet transform analysis: application to simulated and (1)H MRS brain tumor time-domain data. AB - Determination of lactate concentrations in vivo is required in the noninvasive diagnosis, staging, and therapeutic monitoring of diseases such as cancer, heart disease, and stroke. An iterative filtering process based on the continuous wavelet transform (CWT) method in the time domain is proposed to isolate the lactate doublet signal from overlapping lipid resonances and estimate the magnetic resonance spectroscopy (MRS) parameters of the lactate methyl signal (signal amplitude, chemical shift, J-coupling and apparent transverse relaxation time (T*(2))). This method offers a number of advantages over the multiple quantum (MQ) and difference spectroscopy approaches, including: 1) full recovery of the lactate methyl signal, whereas the MQ methods usually detect 50% of the signal intensity; 2) in contrast to MQ methods, the lipid signal is retained together with J-coupling data on the lactate peak; 3) the CWT method is much less sensitive to motion artifacts than difference spectroscopy. Application of the method to simulated and real (1)H MRS data collected from human blood plasma and brain tumors demonstrated that this filter provides accurate estimates of the MRS parameters of the lactate doublet and efficiently removes lipid contributions. PMID- 10800030 TI - 1H and (31)P magnetization transfer studies of hindleg muscle in wild-type and creatine kinase-deficient mice. AB - The authors addressed the hypothesis that interactions with creatine kinase (CK) play a role in the off-resonance magnetization transfer (MT) effect of creatine in skeletal muscle. Toward that aim, (1)H MT studies were done on hindleg muscle in wild-type mice and in transgenic mice, lacking cytoplasmic CK and/or mitochondrial CK. The (1)H MT effect was essentially identical in wild-type muscle and the two single CK knock-out muscles, while moderately decreased in tissue lacking both CK isoforms. (31)P-NMR showed no off-resonance (31)P MT effect in skeletal muscle for PCr in any of the mice, while the enzymatic CK reaction flux was circa 0.2-0.3 sec(-1) in the wild-type muscle and in muscle deficient in mitochondrial CK. The CK enzyme flux was negligible in the other two CK knock-outs. These data suggest that CK plays a minor role in the (1)H MT effect of creatine. Irrespective of the underlying mechanism the creatine MT phenomenon probably has no significant consequences for the thermodynamic availability of total creatine to the CK reaction. PMID- 10800031 TI - Administration and (1)H MRS detection of histidine in human brain: application to in vivo pH measurement. AB - Measurement of histidine in vivo offers the potential for tissue pH measurement using routinely performed (1)H MR spectroscopy. In the brain, however, histidine concentrations are generally too low for reliable measurement. By using oral loading of histidine, this study demonstrates that brain concentrations can be significantly increased, enabling detection of histidine by localized (1)H MR measurements and making in vivo pH measurement possible. In studies carried out on healthy human subjects at 1.5 T, a consistent spectral quality downfield from water was achieved using a PRESS sequence at short echo times. Measurements at different TE values helped to characterize the downfield spectral region. Histidine loading of 400 mg/kg of body weight increased brain histidine levels by approximately 0.8 mM, with maximum histidine concentration reached 4 to 7 hr after consumption. The pH calculated from histidine resonances was 6.96, and a hyperventilation study demonstrated the potential for measuring altered pH. PMID- 10800032 TI - Quantitative MR microscopy of enzymatically degraded articular cartilage. AB - Structural changes in bovine patellar articular cartilage, induced by component selective enzymatic treatments, were investigated by measuring tissue T(2) relaxation at 9.4 T. This MRI parameter was compared with Young's modulus, a measure of elastic stiffness and loadbearing ability of cartilage tissue. Collagenase was used to digest the collagen network and chondroitinase ABC to remove proteoglycans. Polarized light microscopy and digital densitometry were used to assess enzyme penetration after 44 hr of enzymatic digestion. T(2) relaxation in superficial cartilage increased significantly only in samples treated with collagenase. A statistically significant decrease in Young's modulus was observed in both enzymatically treated sample groups. These results confirm that T(2) of articular cartilage is sensitive to the integrity of collagen in the extracellular matrix. Nonetheless, it does not appear to be an unambiguous indicator of cartilage stiffness, which is significantly impaired in osteoarthrosis. PMID- 10800033 TI - Adaptive reconstruction of phased array MR imagery. AB - An adaptive implementation of the spatial matched filter and its application to the reconstruction of phased array MR imagery is described. Locally relevant array correlation statistics for the NMR signal and noise processes are derived directly from the set of complex individual coil images, in the form of sample correlation matrices. Eigen-analysis yields an optimal filter vector for the estimated signal and noise array correlation statistics. The technique enables near-optimal reconstruction of multicoil MR imagery without a-priori knowledge of the individual coil field maps or noise correlation structure. Experimental results indicate SNR performance approaching that of the optimal matched filter. Compared to the sum-of-squares technique, the RMS noise level in dark image regions is reduced by as much as the square root of N, where N is the number of coils in the array. The technique is also effective in suppressing localized motion and flow artifacts. PMID- 10800034 TI - Assessment of regional cerebral blood flow by dynamic susceptibility contrast MRI using different deconvolution techniques. AB - Regional cerebral blood flow (rCBF) was assessed using dynamic susceptibility contrast MRI at 1.5 T. A simultaneous dual FLASH pulse sequence and Gd-DTPA-BMA (0.3 mmol/kg b.w.) were used for examination of 43 volunteers, measuring rCBF in frontal white matter (WM) and in gray matter in the thalamus (GM). Arterial input functions (AIFs) were registered 1) in the carotid artery and 2) in an artery within the GM/WM slice. The measured concentration-vs. -time curve was deconvolved with the AIF using both Fourier Transform (FT) and Singular Value Decomposition (SVD). Relative rCBF was given by the height of the deconvolved response curve. For each volunteer, eight different rCBF maps were calculated, representing different combinations of deconvolution techniques, AIFs, and filters. The average GM-WM rCBF ratios ranged from 2.0-2.2, depending on methodology. Absolute rCBF was 68 +/- 28 ml/(min 100 g) in GM and 35 +/- 13 ml/(min 100g) in WM (mean +/- SD, n = 39). GM-WM rCBF ratios obtained using SVD were 6-10% higher than corresponding ratios obtained using FT. PMID- 10800035 TI - Spin echo entrapped perfusion image (SEEPAGE). A nonsubtraction method for direct imaging of perfusion. AB - Arterial spin-labeled perfusion imaging is increasingly being applied to the study of the brain and other organs. To date, perfusion information has invariably been obtained by subtraction of images with and without spin-labeling of inflowing water. Due to the relatively small amount of blood which enters tissue over a typical inflow period (1-1.5 sec), subtraction errors due to image instability or, in certain circumstances, magnetization transfer effects, can lead to very significant amounts of artifactual image intensity. These problems are avoided in the nonsubtraction method described here. Initially, spins in the imaging slice are selectively saturated, leaving other spins unaffected. A subsequent spin-echo train traps these magnetizations irrespective of flow. Finally, an imaging module generates intensity only from those spins which have entered the imaging slice during the inflow period. A slight modification of the sequence facilitates validation by detecting any contaminating signal in a control image. PMID- 10800036 TI - Robust multiresolution alignment of MRI brain volumes. AB - An algorithm for the automatic alignment of MRI volumes of the human brain was developed, based on techniques adopted from the computer vision literature for image motion estimation. Most image registration techniques rely on the assumption that corresponding voxels in the two volumes have equal intensity, which is not true for MRI volumes acquired with different coils and/or pulse sequences. Intensity normalization and contrast equalization were used to minimize the differences between the intensities of the two volumes. However, these preprocessing steps do not correct perfectly for the image differences when using different coils and/or pulse sequences. Hence, the alignment algorithm relies on robust estimation, which automatically ignores voxels where the intensities are sufficiently different in the two volumes. A multiresolution pyramid implementation enables the algorithm to estimate large displacements. The resulting algorithm is used routinely to align MRI volumes acquired using different protocols (3D SPGR and 2D fast spin echo) and different coils (surface and head) to subvoxel accuracy (better than 1 mm). PMID- 10800038 TI - Motion correction for the quantification of mitral regurgitation using the control volume method. AB - Quantifying mitral regurgitation is difficult because of the complexity of the flow, geometry and motion of the mitral valve. In this paper a MRI compatible phantom was built incorporating a left ventricle and mitral valve motion. Valve motion was obtained using a pneumatic piston. The mitral valve was made regurgitant and the regurgitant volume quantified using a modified control volume method. The modification to the method was the addition of mitral motion correction. This was attained by moving the control volume in unison with the mitral valve and by correcting for this motion in the integration of velocity. This correction was found to be simple, in that it represented the volume swept out by the moving control surface. The measured regurgitant volume was compared to a second MR measurement using a single slice technique, made possible by the tubular construction of the phantom's left atrium. Regression analysis between these two methods produced a regression line of y = 0 + 1.02 x; R = 0.97; standard error of the estimate = 3.47 ml. PMID- 10800037 TI - An analytical SMASH procedure (ASP) for sensitivity-encoded MRI. AB - The simultaneous acquisition of spatial harmonics (SMASH) method of imaging with detector arrays can reduce the number of phase-encoding steps, and MRI scan time several-fold. The original approach utilized numerical gradient-descent fitting with the coil sensitivity profiles to create a set of composite spatial harmonics to replace the phase-encoding steps. Here, an analytical approach for generating the harmonics is presented. A transform is derived to project the harmonics onto a set of sensitivity profiles. A sequence of Fourier, Hilbert, and inverse Fourier transform is then applied to analytically eliminate spatially dependent phase errors from the different coils while fully preserving the spatial encoding. By combining the transform and phase correction, the original numerical image reconstruction method can be replaced by an analytical SMASH procedure (ASP). The approach also allows simulation of SMASH imaging, revealing a criterion for the ratio of the detector sensitivity profile width to the detector spacing that produces optimal harmonic generation. When detector geometry is suboptimal, a group of quasi-harmonics arises, which can be corrected and restored to pure harmonics. The simulation also reveals high-order harmonic modulation effects, and a demodulation procedure is presented that enables application of ASP to a large numbers of detectors. The method is demonstrated on a phantom and humans using a standard 4-channel phased-array MRI system. PMID- 10800039 TI - Correction for acceleration-induced displacement artifacts in phase contrast imaging. AB - The acceleration-induced displacement artifact impairs the accuracy of MR velocity measurements. This study proposes a post processing method for correction of this artifact. Velocity measurements were performed in a flow phantom containing a constriction. Velocity curves were obtained from streamlines parallel to the frequency, phase, and slice directions, respectively. The acceleration-induced displacement artifact was most prominent when the frequency encoding direction was aligned with the flow direction. After correction, velocity assignment improved and a more accurate description of the flow was obtained. In vivo measurements were performed in the aorta in a patient with a repaired aortic coarctation. The correction method was applied to velocity data along a streamline parallel to the frequency encoding direction. The result after correction was a new location of the peak velocity and improved estimates of the velocity gradients. PMID- 10800040 TI - Cerebral perfusion and arterial transit time changes during task activation determined with continuous arterial spin labeling. AB - Perfusion imaging by arterial spin labeling (ASL) can be highly sensitive to the transit time from the labeling site to the tissue. We report the results of a study designed to separate the transit time and perfusion contributions to activation in ASL images accompanying motor and visual stimulation. Fractional transit time decreases were found to be comparable to fractional perfusion increases and the transit time change was found to be the greatest contributor to ASL signal change in ASL sequences without delayed acquisition. The implications for activation imaging with ASL and the arterial control of flow are discussed. PMID- 10800041 TI - The effect of bulk susceptibility on murine snapshot imaging at 7.0 T: a comparison of snapshot imaging techniques. AB - The sensitivity of several single-shot imaging techniques to local field gradients (LFGs) generated by discontinuities in bulk magnetic susceptibility (BMS) were compared in mouse brain at 7.0 T. At high field, differences in BMS can cause substantial signal attenuation and image distortion. Because susceptibility effects scale with B(o), mouse brain single-shot imaging at high field is particularly susceptible to the effects of LFGs. The spin-echo techniques GRASE and RARE were found to have a decreased sensitivity to LFGs compared to echo-planar imaging (EPI). Images obtained using EPI and SE-EPI exhibit severe signal attenuation in regions of high LFGs such as near air-tissue interfaces and at the brain edges. In applications such as functional MRI and diffusion MRI, GRASE and RARE are likely to provide more comprehensive whole brain coverage in mouse brain at high field than EPI techniques, which are likely to image regions of the brain with strong LFGs with low signal to noise, reducing the probability of detecting significant physiologically based changes. PMID- 10800042 TI - Contribution of trifluoperazine metabolites to the in vivo (19)F NMR spectrum of rat brain. AB - Fluorine-19 NMR spectra were acquired from extracts of tissues from heads of rats given the antipsychotic drug trifluoperazine (TFP). Contributions to the in vivo (19)F spectra from tissues other than brain were negligible. The in vivo (19)F resonance at -62.3 ppm from CCl(3)F consisted of 6-8 resolved resonances in vitro. Some in vitro resonances were assigned to previously identified TFP metabolites. Multiple resonances in vitro partially explain the relatively large line width seen in vivo for TFP. Unidentified metabolites were observed at about 74 to -75 ppm in a number of spectra of extracts of brain and muscle. PMID- 10800043 TI - Effect of respiratory phases on MR lung signal intensity and lung conspicuity using segmented multiple inversion recovery turbo spin echo (MIR-TSE). AB - The purpose of this study was to determine the effect of respiratory phase on signal intensity of the lung. Lung images were obtained from eleven healthy human volunteers using a multiple inversion recovery segmented turbo spin echo sequence (MIR-TSE). MIR exploits the difference in T(1) between different tissues to effectively null signal contributions from fat and muscle for improved visualization of the lung. The volunteers were asked to perform breath-holding on end inspiration or end expiration. There was a significant decrease in signal intensity of the lung with average SNR of 7.3 +/- 0.9 vs. 14.4 +/- 0.8 for coronal slices, and 9.5 +/-1.5 vs. 16.0 +/-2.4 for sagittal breath-hold images acquired during end inspiration compared with end expiration. It is concluded that MRI of the lungs should be performed during end expiration in order to optimize image quality. PMID- 10800044 TI - Contrast optimization of fluid-attenuated inversion-recovery (FLAIR) MR imaging in patients with high CSF blood or protein content. AB - After surgical resection of a brain tumor or infection of the cerebrospinal fluid (CSF), elevated levels of blood by-products or protein contaminations are seen in the patient's CSF spaces. In fast fluid-attenuated inversion-recovery (FLAIR) imaging CSF signal is nulled by an appropriate choice of the inversion recovery time TI to improve the contrast between tissue structures adjacent to CSF-filled volumes. With contaminated CSF, however, the longitudinal relaxation time T(1) may change significantly, which results in an incomplete suppression in the FLAIR images, if standard inversion times are used. In this work, a fast single-voxel T(1) measurement pulse sequence with integrated T(1) calculation that allows determination the optimal TI value in 15 sec is presented. The method was tested in five patients after surgical resection of a brain tumor, where FLAIR MRI with and without contrast agent was performed to identify remaining tumor fragments at the margin of the resection cavity. PMID- 10800045 TI - Comparison of the temporal response in perfusion and BOLD-based event-related functional MRI. AB - Event-related functional MRI (ER-fMRI) based on both blood oxygen level-dependent (BOLD) contrast and perfusion contrast has been recently developed to study human brain activation due to brief stimulation. In this report, both BOLD- and perfusion-based ER-fMRI were directly compared using repeated single-trial, short visual stimulation (1 sec) in six human volunteers. The results show that the cerebral blood flow change reached a maximum approximately 1 sec earlier than the BOLD signal change (4.2 +/- 0.2 sec vs. 5.1 +/- 0.2 sec after the stimulation, P < 0.05). The full width at half maximum of the hemodynamic response measured by perfusion was not significantly different from that measured with BOLD (5.1 +/- 0.6 sec vs. 5.9 +/- 0.6 sec). A positive linear correlation was found between the maximum perfusion and maximum BOLD signal changes (r = 0. 77, P = 0.07). PMID- 10800046 TI - PURR-TURBO: a novel pulse sequence for longitudinal relaxographic imaging. AB - A novel pulse sequence based on a segmented phase-encoding scheme for measuring the longitudinal relaxation time (T(1)) value of an NMR signal in an imaging context is introduced. This pulse sequence is a hybrid version of the "single shot" and "one-shot" inversion recovery (IR) snapshot-FLASH methods. These are also known as "multipoint" IR techniques. The new sequence presented here collects multiple k-space rows at each time point during magnetization recovery, rather than the entire dataset (as in a "single-shot" method) or just one row (as in a "one-shot" method). Thus, it reduces the scanning time without significant sacrifice of the small sampling time advantage of the one-shot IR snapshot-FLASH methods. Furthermore, this approach does not require a high-performance gradient system. Here, we demonstrate that a single slice human brain (1)H(2)O T(1) map with a nominal in-plane resolution of less than (1 mm)2can be obtained at 4 T in about 4 min. Published 2000 Wiley-Liss, Inc. PMID- 10800047 TI - Sampling and analysis of individual particles by aerosol mass spectrometry. AB - Over the past decade, aerosol mass spectrometry has developed into a powerful method for characterizing individual particles in air. Recent advances in the design of inlets and mass spectrometers have extended the size range of particles that can be analyzed. In this tutorial, fundamental aspects of particle motion in sampling inlets are introduced. Basic experimental configurations for achieving a high analysis rate and the ability of laser ablation to provide chemical composition information are reviewed. An example of the use of this technology to study atmospheric phenomena is also presented. Significant opportunity exists for designing new experiments at the interface of aerosol mass spectrometry and conventional molecular mass spectrometry. PMID- 10800048 TI - Characterization of phosphatidylethanolamine as a lithiated adduct by triple quadrupole tandem mass spectrometry with electrospray ionization. AB - Structural characterization of the glycerophosphoethanolamine (GPE) molecule as a lithiated adduct ion by collisionally activated dissociation (CAD) tandem mass spectrometry with electrospray ionization is described. Abundant fragment ions reflecting polar head group and fatty acid constituents were observed in the product ion spectrum of GPE, which permits an unambiguous structural determination, including the regiospecificity of fatty acyl substituents. The pathways leading to the formation of fragment ions are proposed. The suggested mechanisms are supported by the tandem mass spectra of various deuterated analogs and source CAD of GPE followed by CAD tandem mass spectrometry. Identification of GPE molecular species and specific GPE subclasses in a biological mixture by tandem mass spectrometry with various constant neutral loss scannings is also described. PMID- 10800049 TI - Electrospray ionization mass spectrometry of stable nitroxide free radicals and two isoindoline nitroxide dimers. AB - Electrospray ionization (ESI) mass spectra were recorded for a range of substituted isoindoline nitroxides, two isoindoline nitroxide dimers and two piperidinyl nitroxides. In all cases the dominant molecular species arise from oxidation rather than protonation, an unusual process in ESI. Fragment ion spectroscopy was used to establish fragmentation mechanisms for the nitroxides under ESI conditions. PMID- 10800050 TI - Identification of non-covalent structure in apocytochrome c by hydrogen exchange and mass spectrometry. AB - Apocytochrome c, the in vivo precursor to active cytochrome c, was analyzed by amide hydrogen exchange and mass spectrometry to search for fixed, non-covalent structure. The protein was incubated in H(2)O at pH 3.3 or 6.7 for various times, then exposed to D(2)O to initiate isotope labeling of unfolded regions. Following acid quenching of hydrogen exchange, the labeled apocytochrome c was digested with pepsin into fragments that were analyzed by directly coupled high performance liquid chromatography/electrospray ionization mass spectrometry. The intermolecular distribution of deuterium and the deuterium levels in structurally distinctive populations were determined from the mass spectra of the peptic fragments. Spectra of peptic fragments derived from apocytochrome c incubated at pH 3.3 had single envelopes of isotope peaks with masses indicating that all of the amide hydrogens had been replaced with deuterium. These results showed that apocytochrome c at pH 3.3 offered little resistance to hydrogen exchange, indicating that it was unfolded with little fixed structure. However, mass spectra of peptic fragments including residues 81-94 of apocytochrome c incubated at pH 6.7 had two envelopes of isotope peaks, indicating that one population was unfolded and the other population was highly structured in this region. Mass spectra of peptic fragments including residues N-terminal to residue 81 indicated that this region of the protein remained unfolded with little fixed structure at pH 6.7. PMID- 10800051 TI - On-line monitoring by membrane introduction mass spectrometry of chlorination of organics in water. Mechanistic and kinetic aspects of chloroform formation. AB - Chloroform formation during the chlorination of simple organic molecules modeling humic substances, such as phenol and di- and trihydroxybenzenes, was studied by on-line membrane introduction mass spectrometry (MIMS). Under the reaction conditions employed, chloroform was rapidly formed from 1,3-dihydroxybenzene, 1, 4-dihydroxybenzene, phenol and 1,2,3-trihydroxybenzene with yields of 17, 13, 7 and 5%, respectively. With the exception of aniline, which afforded a 17% chloroform yield, non-phenolic compounds, such as nitrobenzene, chlorobenzene, toluene, benzene and cyclohexanol, furnished low yields. Mechanistic studies showed that phenol is chlorinated consecutively and produces initially chlorophenol. It is suggested that chloroform might be formed mainly from chlorinated 3, 5-cyclohexadienone-type intermediates. MIMS was also used to determine the reaction rates and to study the kinetics of the chlorination. A good Hammett linear correlation for an electrophilic substitution mechanism was found for the compounds C(6)H(5)X (X = NH(2), OH, CH(3), H, Cl and NO(2)). PMID- 10800052 TI - Fully automated determination of eserine N-oxide in human plasma using on-line solid-phase extraction with liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - A sensitive and entirely automated solid-phase extraction/liquid chromatography/electrospray ionization tandem mass spectrometric (SPE/LC/ESI MS/MS) method was developed and validated for the determination of eserine N oxide (ENO), a cholinesterase inhibitor-like physostigmine in human plasma, for use in pharmacokinetic studies. ENO is light-sensitive and the use of a fully on line process increased the reliability of the assay. Plasma samples previously mixed with neostigmine bromide to prevent in vitro degradation, and tacrine as internal standard (IS), were directly injected into the SPE/LC/ESI-MS/MS system. MS software piloted the overall system. MS/MS detection of ENO and the IS was performed in the positive ion ESI mode using multiple reaction monitoring. The linear calibration curve for ENO ranged from 25 pg ml(-1) to 12.5 ng ml(-1). The limit of quantitation was 25 pg ml(-1) with 250 microl of plasma injected. Precision, accuracy and stability tests were within the acceptable range and just one analyst is required to analyze 50 unknown samples a day five days per week, from the preparation of the samples (i.e. thawing and centrifugation) to data processing. A pilot pharmacokinetic study in three healthy volunteers treated with 4.5 mg of ENO (Geneserine3((R))) showed that the method was suitable for pharmacokinetic studies in humans. PMID- 10800053 TI - Chemical ionization mass spectra of acetals of beta-D-glycopyranosylnitromethanes AB - O-Isopropylidene and O-benzylidene acetals of common 2, 6-anhydro-1-deoxy-1 nitroalditols (beta-D-glycopyranosylnitromethanes) derived from D-glucose, D galactose and D-mannose were studied by chemical ionization mass spectrometry (CIMS) using methane, isobutane, ammonia or pyridine as reaction gas. Production of [M+H](+) adduct ions dominates in the case of methane or isobutane possessing proton affinity values PA = 552 or 683 kJ mol(-1), respectively. The collision induced dissociation time-of-flight product ion spectra of [M+H](+) ions differ characteristically according the stereochemical arrangement of the pyranoid ring. These differences can be helpful when assigning stereochemical arrangements for the pyranoid ring. The dominant process in ammonia (PA = 853 kJ mol(-1)) CIMS for most of the compounds studied is the production of the cluster ions [M+NH(4)](+). The cluster [M+pyridineH](+) ions are observable only for substances possessing the O-benzylidene group (PA of pyridine = 924 kJ mol(-1)). Copyright 2000 John Wiley & Sons, Ltd. PMID- 10800054 TI - Comparison of different derivatization approaches for mercury speciation in biological tissues by gas chromatography/inductively coupled plasma mass spectrometry. AB - A novel interface design for coupling gas chromatography and inductively coupled plasma mass spectrometry (GC/ICP-MS) was used to perform mercury speciation in biological tissues. Three derivatization approaches were optimized and compared for this purpose: anhydrous butylation using a Grignard reagent, aqueous ethylation by means of NaEt(4)B and aqueous propylation with NaPr(4)B. The last reagent was synthesized in the laboratory as it is not commercially available. Detection limits obtained by GC/ICP-MS ranged between 100 and 200 fg (as absolute mass) for methylmercury and between 500 and 600 fg for inorganic mercury using a 1 microl injection. Quantification of methyl- and inorganic mercury was carried out by resorting to aqueous calibration, using ethylmercury as internal standard for both propylation and butylation derivatization techniques. For ethylation procedures, a methylpropylmercury solution was used as internal standard. The absence of transmethylation during sample preparation was checked using a 97% enriched (202)Hg inorganic standard. The accuracy of the three derivatization approaches was evaluated by the analysis of the certified reference material DOLT 2 (dogfish liver) from the National Research Council of Canada and certified for methylmercury, with satisfactory results. PMID- 10800055 TI - On-probe sample purification of lipids for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 10800056 TI - Current awareness AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of mass spectrometry. Each bibliography is divided into 11 sections: 1 Books, Reviews & Symposia; 2 Instrumental Techniques & Methods; 3 Gas Phase Ion Chemistry; 4 Biology/Biochemistry: Amino Acids, Peptides & Proteins; Carbohydrates; Lipids; Nucleic Acids; 5 Pharmacology/Toxicology; 6 Natural Products; 7 Analysis of Organic Compounds; 8 Analysis of Inorganics/Organometallics; 9 Surface Analysis; 10 Environmental Analysis; 11 Elemental Analysis. Within each section, articles are listed in alphabetical order with respect to author (2 Weeks journals - Search completed at 16th Feb. 2000) PMID- 10800057 TI - The effect of busulphan on the pharmacokinetics of cyclophosphamide and its 4 hydroxy metabolite: time interval influence on therapeutic efficacy and therapy related toxicity. AB - Busulphan and cyclophosphamide (Bu/CP) are widely used in preparative regimens for bone marrow transplantation. Many studies have shown a wide variation in busulphan pharmacokinetics. Moreover, higher rates of liver toxicity were reported in Bu/CP protocols than in a total body irradiation (TBI)-containing regimen. In the present paper we investigated the effect of the time interval between the last dose of busulphan and the first dose of cyclophosphamide on the pharmacokinetics of CP and its cytotoxic metabolite 4-hydroperoxycyclophosphamide (4-OHCP). Thirty-six patients undergoing bone marrow transplantation (BMT) were included in the study. We also investigated the occurrence of veno-occlusive disease, mucositis and graft-versus-host disease. Ten patients conditioned with CP followed by TBI served as a control group (TBI). Twenty-six patients were conditioned with Bu/CP. The patients received Bu (1 mg/kg x 4 for 4 days), followed by CP (60 mg/kg for 2 days) administered as a 1-h infusion. Patients received their CP therapy either 7-15 h (group A, n = 12) or 24-50 h (group B, n = 14) after the last dose of Bu. None of the patients were given phenytoin or any other drug known to enhance CP metabolism. The administration of CP less than 24 h after the last dose of Bu resulted in: (1) a significantly (P = 0.003) lower clearance for cyclophosphamide was observed in group A (0.036 l/h/kg) compared to 0.055 and 0.055 l/h/kg, in the B and TBI groups, respectively; (2) significantly (P = 0.002) longer elimination half-life in group A (10.93 h) than in groups B and TBI (6.87 and 7.52 h, respectively); (3) significantly (P < 0.001) lower exposure to the cytotoxic metabolite (4-OHCP), expressed as the ratio AUC4 OHCP/AUCCP, in group A (0.0053) than that obtained in group B (0.013) and group TBI (0.012); (4) the patients in group A had a significantly (P < 0.05) higher incidence of VOD (seven of 12) than the other groups, B and TBI (2/14 and 1/10, respectively); and (5) mucositis was significantly higher in group A patients (8/12), being seen in only one patient in group B and none in the TBI group. The present study has shown that the interval between busulphan and cyclophosphamide administration can negatively affect the pharmacokinetics of cyclophosphamide and its cytotoxic metabolite. We conclude that the timing of CP administration must be considered in order to improve drug efficacy and reduce conditioning-related toxicity. PMID- 10800059 TI - Single vs twice daily G-CSF dose for peripheral blood stem cells harvest in normal donors and children with non-malignant diseases. AB - The optimal dose and schedule of G-CSF for mobilization of peripheral blood stem cells (PBSC) is not well defined. G-CSF mobilization was performed in a group of healthy donors and paediatric patients for autologous back-up before receiving allogeneic stem cell transplant. Seventeen consecutive subjects who received G CSF at 5 microg/kg/dose twice daily (group A) were compared with a historical control group of 25 subjects who received a single daily dose of 10 microg/kg/day G-CSF (group B). Double blood volume apheresis for PBSC collection was started on day 5. G-CSF was continued and apheresis repeated until the targeted CD34+ cell dose was achieved. Both groups were comparable for sex, age, body weight and reason for PBSC collection. Over two-thirds of the subjects in both groups were less than 16 years of age. The G-CSF priming and apheresis were well tolerated. When the first day apheresis products were analyzed, group A resulted in significantly higher yield of total nucleated cells (5.91 vs 3.92 x 108/kg, P = 0. 013), mononuclear cells (5.73 vs 3.92 x 108/kg, P = 0.017), CD34+ cells (2.80 vs 1.69 x 106/kg, P = 0.049) and colony-forming units (107 vs 54 x 104/kg, P = 0.010) as compared with group B. We conclude that the two dose schedule is more efficient in mobilizing PBSC in normal donors and children with non-malignant diseases. This approach may reduce the number of aphereses required and thus reduce the transplant cost. PMID- 10800058 TI - An evaluation of engraftment, toxicity and busulfan concentration in children receiving bone marrow transplantation for leukemia or genetic disease. AB - Autologous recovery is a major problem with busulfan as a marrow ablative agent in conditioning children for allogeneic BMT. Data suggest the average concentration of busulfan at steady state (Bu Css) is critical for successful engraftment. We prospectively evaluated busulfan pharmacokinetics in 31 children (age 0.6-18 years) with AML (n = 9), and non-malignant diseases (n = 22) receiving HLA-closely matched (sibling, parent, unrelated) donor grafts. Blood samples were obtained following dose 1 and 13 of a standard 16 dose, 4-day regimen. The busulfan dose varied from 14 to 20 mg/kg. Patients received cyclophosphamide 200-240 mg/kg; 22/31 received 80-90 mg/kg of ATG. Eight patients failed to engraft (26%). ATG did not appear to influence engraftment (P = 0.38). Bu Css levels <600 ng/ml correlated with autologous recovery/mixed chimerism (P = 0.018). There were no graft failures in patients with a Bu Css >600 ng/ml. A correlation between Bu Css levels and regimen-related toxicity (RRT) was not identified for grade 2 or higher toxicities, only 1/31 had a Bu Css >900 ng/ml. Our data support the use of pharmacokinetic monitoring of busulfan. PMID- 10800060 TI - Consolidation with a busulfan-containing regimen followed by stem cell transplantation in infants with poor prognosis stage 4 neuroblastoma. AB - Although infants with stage 4 neuroblastoma (NB) usually have a good prognosis, metastatic relapses after 1 year of age and amplification of the N-myc oncogene are established poor prognostic factors. In order to improve the survival of patients with such high-risk factors, we performed consolidation with a busulfan (600 mg/m2)-melphalan (140 mg/m2)-containing regimen followed by autologous stem cell transplantation (SCT). From 1986 to 1998, 12 patients were treated according to this strategy. Their median age at diagnosis was 9 months (1-11). Consolidation was performed after a metastatic relapse in five children, because of persistent bone metastases in one and as first-line consolidation in six patients whose tumor exhibited N-myc amplification. The 5-year EFS rate is 64. 5% (36-85%) with a median follow-up of 92 months (20-126). One toxicity-related death occurred in a very heavily pretreated patient. Hepatic veno-occlusive disease was the major side-effect that occurred in nine of 12 children. This busulfan-melphalan combination appears to dramatically improve the prognosis of these high-risk infants with metastatic NB. Given its high toxicity, indications for this consolidation must be restricted to high-risk infants and a lower dose of busulfan (480 mg/m2) is recommended in children weighing less than 10 kg. Bone Marrow Transplantation (2000) 25, 937-942. PMID- 10800061 TI - Poor outcome in children with refractory/relapsed leukemia undergoing bone marrow transplantation with mismatched family member donors. AB - The utility of bone marrow transplantation for childhood leukemia in patients unable to achieve a remission prior to transplant is controversial. To address this issue, we analyzed a subset of patients with advanced leukemia entered on prospective transplant trials at our hospital. Fifty-eight patients with ALL or AML (age 1-19) were identified. They had failed standard chemotherapy and were in relapse (22 in 1st, 27 in 2nd, three in 3rd, and three in 4th) or had never achieved an initial remission (three) at the time of transplant. Fifty-two patients received marrow from mismatched family members (haplo or DR-identical), while six received marrow from matched siblings. Most patients received myeloablative therapy consisting of total body irradiation, etoposide, cyclophosphamide, and cytosine arabinoside. Marrow from mismatched donors was T cell depleted. Only one of 52 patients transplanted with a mismatched donor survived long-term while three of six patients transplanted in relapse with a fully matched sibling donor are alive 6-10 years post BMT. The major causes of death were infection (39%) and relapse (28%). Acute GVHD grade III-IV was noted in 7% of patients. A comparable group of patients with leukemia transplanted at our center in remission using similarly mismatched family member donors (haplo or DR-identical) had an event-free survival of 28%. In conclusion, our data suggest that BMT utilizing mismatched family member donors is a poor option for patients in relapse at the time of transplant. New treatment strategies need to be developed to effectively manage these patients. PMID- 10800062 TI - Acute lymphoblastic leukaemia-type intensive chemotherapy to eliminate minimal residual disease after high-dose melphalan and autologous transplantation in multiple myeloma - a phase I/II feasibility and tolerance study of 17 patients. AB - Aiming to target the minimal residual disease in patients with multiple myeloma, a phase I/II single centre study was undertaken for feasibility and tolerance of intensive acute lymphoblastic leukaemia consolidation chemotherapy (ALL-IC) as part of a strategy for post-transplant consolidation targeted at pre-B cells. Seventeen newly diagnosed patients with myeloma (median age 55 years; 30-65) were initially treated with courses of infused cyclophosphamide, vincristine, adriamycin and methylprednisolone (C-VAMP) followed by melphalan 200 mg/m2(HDM) and peripheral blood stem cell rescue (PBSC). Forty-seven percent were in CR and the rest in PR after HDM. ALL-IC consisted of vincristine, daunorubicin, etoposide, cytarabine, 6-thioguanine and prednisolone given over 5 days. All patients became neutropenic (<0.5 x 109/l) at a median of 10 days (4-18) and one of the 17 patients (5.8%) died 15 days post ALL-IC of sepsis. A further four have died of relapse with an overall survival (OS) of 67% at 4 years. Two of nine patients in PR at the time of ALL-IC achieved CR. Matched-pair analysis of 34 control patients shows no difference for OS and event-free survival between ALL IC and controls. We conclude that ALL-IC given to myeloma patients after HDM/PBSC is as safe as when used in ALL and warrants further assessment in randomised trials for myeloma. PMID- 10800063 TI - Prognostic factors for the clinical outcome of patients with follicular lymphoma following high-dose therapy and peripheral blood stem cell transplantation (PBSCT). AB - This is a report on 111 patients with advanced stage follicular lymphoma who where autografted using PBSC. Seventy patients were enrolled in first remission, whereas 41 were treated in second or higher remission. High-dose therapy consisted of total body irradiation plus cyclophosphamide in 103 patients, while eight patients received BEAM (carmustine, etoposide, cytosine-arabinoside, melphalan). Autografts contained 8.1 +/- 0.6 x 106 CD34+ cells/kg body weight. At a median follow-up of 44.2 months from PBSCT (range 4.9-77.4 months), 93 patients are alive, with a probability of overall and relapse-free survival (RFS) of 83% and 64%, respectively. A significantly higher probability of relapse was associated with male gender, involvement of more than eight lymph node areas, extra-nodal manifestations other than bone marrow and PBSCT performed in second or higher remission. In the latter group of patients, previous radiotherapy was associated with poor prognosis. The relevance of chemosensitivity as a prognostic factor was reflected by a better RFS in patients who had achieved complete remission at the time of PBSC mobilization. In a multivariate analysis, involvement of eight or more lymph nodes and high-dose therapy performed in second or higher remission were independent prognostic factors. PMID- 10800064 TI - New oral formulation of cyclosporin A (Neoral) pharmacokinetics in allogeneic bone marrow transplant recipients. AB - Cyclosporin A (CsA) absorption is variable in bone marrow transplant (BMT) patients compromising the efficacy of graft-versus-host disease prevention. Neoral, a new microemulsion formulation of CsA which has an improved bioavailability, increases intestinal absorption of the drug with less variable pharmacokinetic parameters in non-BMT patients. In order to predict the best dosage of Neoral when patients are switched from i.v. to oral administration we performed a randomised study comparing two oral doses, either the same or twice the last i.v. dose used after BMT. Fourteen adults were randomised around day 25 after BMT. Whole blood CSA concentrations were measured 2 and 12 h after the oral administration of Neoral on days 0, 7 and 14 to determine residual and maximum concentration, and modified whenever necessary to maintain blood level CsA concentration within therapeutic range (150-250 ng/ml). We found that patients who received twice the last i.v. dose had better concentrations than patients from the other group while toxicity was identical in both groups. We conclude that doubling the last i.v. dose during the switch to oral administration of Neoral gives the best therapeutic range concentration and should be recommended for graft-versus-host prevention. PMID- 10800065 TI - Disseminated toxoplasmosis following T cell-depleted related and unrelated bone marrow transplantation. AB - More than 95% of reported cases of disseminated toxoplasmosis following BMT have occurred following an unmodified transplant. Most have been fatal, diagnosed at autopsy and without antemortem institution of specific therapy. From 1989 to 1999, we identified 10 cases of disseminated toxoplasmosis, in 463 consecutive recipients of a T cell-depleted (TCD) BMT. Transplants were from an unrelated donor (n = 5), an HLA-matched sibling (n = 4) or an HLA-mismatched father (n = 1). In 40%, both the donor and recipient had positive IgG titers against T. gondii pre-transplant; in 30%, only the recipient was sero-positive. Three recipients of an unrelated TCD BMT developed toxoplasmosis despite both donor and host testing negative pretransplant. All 10 patients presented with high grade fever. CNS involvement ultimately occurred in seven patients, with refractory respiratory failure and hypotension developing in nine. Eight of 10 cases were found only at autopsy, involving the lungs (n = 7), heart (n = 5), GI tract (n = 5), brain (n = 8), liver and/or spleen (n = 5). The only survivor, treated on the day of presentation with fever and headache, was diagnosed by detection of T. gondii DNA by polymerase chain reaction (PCR) performed on the blood and spinal fluid. This study demonstrates the similar incidence of toxoplasmosis following TCD BMT and that reported post T cell-replete BMT, and underscores the need for rapid diagnostic tests in an effort to improve outcome. PMID- 10800066 TI - Bronchoalveolar lavage in the diagnosis of pulmonary complications of bone marrow transplant patients. AB - Bronchoalveolar lavage (BAL) has proved valuable in the diagnosis of pulmonary complications in immunosuppressed patients. We evaluated the diagnostic yield of BAL in pulmonary complications in bone marrow transplantation (BMT) recipients. We reviewed sequentially the records of 89 patients during an 18-month period. BAL was diagnostic in 42 patients (47%). The most common pulmonary complication diagnosed by BAL was diffuse alveolar hemorrhage (n = 15); followed by bacterial pneumonia (n = 10), respiratory syncytial virus (n = 8), aspergillosis (n = 6), Pneumocystis carinii pneumonia (n = 5), cytomegalovirus (CMV) (n = 4), and others (n = 4). The final diagnoses in the BAL non-diagnostic group were: bacterial pneumonia (n = 6), CMV (n = 6), idiopathic pneumonia syndrome (n = 5), cancer recurrence (n = 4), cardiogenic pulmonary edema (n = 4), and others (n = 9). We conclude that BAL is a useful diagnostic tool in BMT-related pulmonary complications. PMID- 10800067 TI - Encouraging results in the treatment of haemorrhagic cystitis with estrogen - report of 10 cases and review of the literature. AB - Haemorrhagic cystitis (HC) after allogeneic haematopoietic stem cell transplantation (HSCT) or high-dose cyclophosphamide (CP) chemotherapy is a severe side-effect and can cause significant morbidity and mortality. In this report, we describe the clinical courses of 10 patients with HC and review the literature. The patients were treated with oral conjugated estrogen in an attempt to improve severe haemorrhagic cystitis. In seven patients positive effects were seen, haematuria resolved in all, but residual symptoms of dysuria remained for longer periods. In one patient application of estrogen was interrupted because of hepatotoxicity. Two patients failed all treatment modalities including oral estrogen because of terminal illness. We conclude that in the management of HC the administration of oral conjugated estrogen should be considered. PMID- 10800068 TI - Transjugular intrahepatic portosystemic shunt (TIPS) for severe veno-occlusive disease of the liver following bone marrow transplantation. AB - Severe veno-occlusive disease (VOD) of the liver is a leading cause of mortality after bone marrow transplantation (BMT). Vascular and parenchymal injuries account for acute portal hypertension and liver failure is frequently present. We describe the results of transjugular intrahepatic portosystemic shunt (TIPS) for the management of VOD after BMT. TIPS was performed in 10 patients with histologically proven severe VOD. Portal hypertension was controlled by TIPS in all patients (mean hepatic venous pressure gradient before, 20 +/- 11 vs 6 +/- 5 mm Hg after TIPS, P < 0.01) without technical complications. Five patients with rapidly worsening VOD died within 10 days of TIPS without any improvement. The five remaining patients with less advanced disease showed improvement in various clinical and biological parameters. Four patients subsequently died. The lone survivor continues to do well with resolution of VOD 6 months after TIPS. TIPS can be performed safely and controls portal hypertension in VOD after BMT. Arguments from the present series and from eight previously reported cases favour earlier application of TIPS to obtain improved overall survival. Bone Marrow Transplantation (2000) 25, 987-992. PMID- 10800069 TI - N-acetylcysteine for hepatic veno-occlusive disease after allogeneic stem cell transplantation. AB - Three patients developed veno-occlusive disease of the liver (VOD) after allogeneic stem cell transplantation. On the day after diagnosis, N acetylcysteine (NAC) was given, initially in loading doses and thereafter 50-150 mg/kg/day for 12 to 31 days. The maximum bilirubin levels were 137, 58 and 138 mmol/l in the three patients, respectively. After the introduction of NAC, bilirubin, aspartate aminotransferase, sIL-2 receptor and IL-8 decreased. All three patients achieved normal bilirubin levels and prothrombin times. To conclude, NAC may be useful for treatment of VOD. PMID- 10800070 TI - Cost-effectiveness of CD34+ dose in peripheral blood progenitor cell transplantation for non-Hodgkin's lymphoma patients: a single centre study. AB - Intensive high-dose chemotherapy with peripheral blood progenitor cell (PBPC) transplantation is a common strategy for aggressive non-Hodgkin's lymphomas (NHL). A retrospective cost-effectiveness analysis of CD34+ cell dose was carried out. Between 1994 and 1998, 28 patients were included. Efficacy was measured by the length of aplasia. Data collection concerned the period from graft day until discharge from hospital, and the post-graft period until graft day +100. Patients transplanted using a cell dose greater than 5 x 106/kg were found to have a faster hematological recovery. Average length of post-graft hospitalization was shorter and fewer blood products were required for patients with more than 5 x 106/kg CD34+ cells transplanted. Hospitalization was the major cost driver. A large reduction in procedure cost was obtained with a CD34+ cell count higher than 5 x 106/kg (-US$2740, -11%). This difference was directly related to hospitalization (-US$860) and platelet units transfused (-US$1,340). A sensitivity analysis showed the robustness of results. Our findings indicated that a CD34+ cell dose higher than 5 x 106/kg was more cost-effective than a lower dose in NHL patients. The collection of 5 x 106/kg CD34+ cells appeared necessary to optimize the PBPC procedure. PMID- 10800071 TI - Acute abdomen without cutaneous signs of varicella zoster virus infection as a late complication of allogeneic bone marrow transplantation: importance of empiric therapy with acyclovir. AB - Two patients complained of severe abdominal pain as the first sign of varicella zoster virus infection about 1 year after allogeneic BMT. In case 1, eruptions, found on the face and chest on admission, became vesicular and dispersed on the third hospital day. Though acyclovir (ACV) was immediately started, he died on the fourth day. In case 2, skin rash was never observed during the clinical course. Laparotomy on the third hospital day revealed many hemorrhagic spots on the liver surface and mucous membrane of the upper GI tract, indicating disseminated visceral disease. Empiric therapy with ACV was successful. PMID- 10800072 TI - Dental manifestations in severe combined immunodeficiency following bone marrow transplantation. AB - Severe combined immunodeficiency (SCID) is a rare primary immunodeficiency disorder with an estimated overall frequency of 1 in 75 000 live births. Bone marrow transplantation is the only curative treatment available. Using T cell depleted HLA non-identical bone marrow requires preconditioning with a short course of cytotoxic chemotherapy. We report severe dental developmental anomalies in three such patients under long-term follow up. PMID- 10800073 TI - Radiation-induced bone sarcoma following total body irradiation: role of additional radiation on localized areas. AB - A 44-year-old patient who had had acute monoblastic leukemia developed an osteosarcoma of the pelvic bones 5 years after an allogeneic bone marrow transplant from his HLA-identical sister. He had additionally received superficial cutaneous radiation of the legs and pelvis, over the 3 weeks prior to total body irradiation (TBI), because of cutaneous leukemic lesions. The tumor was a fibrohistiocytomatous osteogenic sarcoma. The first lesion was in the right ilium, and a second lesion appeared 18 months later, symmetrically on the left ilium. Despite treatment, the patient died from metastases. At the time of diagnosis of radiation-induced sarcoma, the patient was free of leukemia and had several risk factors already reported to favor the development of solid tumors in stem cell recipients. These include acute leukemia, TBI and graft-versus-host disease. As he developed symmetrical lesions of the pelvic bone, and because of the histology of the radiation-induced tumor, we assumed that the additional radiation of the skin prior to TBI may have contributed to the pathogenesis of this malignant fibrous histiocytoma. Therefore, the risk/benefit ratio should be carefully considered in unusual indications. These patients should benefit from a close follow-up of the superimposed areas. PMID- 10800074 TI - Severe hypophosphatemia during hematopoietic reconstitution after allogeneic peripheral blood stem cell transplantation. AB - A patient suffering from acute myeloid leukemia (FAB M5a) received a PBSC allograft from a matched, related donor. On day 13 after transplantation severe hypophosphatemia (0.21 mmol/l) was first noted which persisted irrespective of intravenous phosphate administration, and within 2 days reached concentrations below 0.13 mmol/l. After repeated phosphate substitution serum phosphate returned to 1.40 mmol/l on day 17. Phosphate in urine, and calcium in serum were recorded as unchanged throughout. Clinical signs and symptoms due to severe hypophosphatemia were not observed except for paresthesia in the lower extremities. The precipitous fall in serum phosphate coincided with hematopoietic reconstitution as reflected by a steep rise in leukocyte count from 0.08 x 109/l on day 10 to 5. 94 x 109/l on day 15 after transplantation. Thus, isolated hypophosphatemia was likely the result of excessive cellular phosphate uptake during hematopoietic reconstitution. Electrolyte monitoring after PBSCT should include serum phosphate to identify the hypophosphatemia associated with hematopoietic recovery. PMID- 10800075 TI - Perturbation of membrane microdomains reduces mitogenic signaling and increases susceptibility to apoptosis after T cell receptor stimulation. AB - Acid sphingomyelinase-deficient (asmase-/-) mice generated by gene targeting abundantly store sphingomyelin in the reticuloendothelial system of liver, spleen, bone marrow, and in brain. Liver cells of asmase-/- mice accumulate sphingomyelin and glycosphingolipids in purified lipid bilayers of microsomes, Golgi, and the plasma membrane, but cholesterol is depleted in the plasma membrane. Detergent-insoluble glycolipid-enriched membrane microdomains (GEM) can be isolated from hepatocytes, embryonic fibroblasts, and splenocytes of wild type, but not of asmase-/- mice, by sucrose gradient density centrifugation. Lck and other Src-family kinases are reduced in isopycnic fractions of asmase-/- splenocytes compared to GEM-containing fractions of wild-type cells. The proliferation of asmase-/- T lymphocytes is reduced, whereas their susceptibility to Fas-induced apoptosis is increased after T cell receptor (TCR) stimulation. TNF receptor I signaling remains unimpaired. The perturbation of GEM impairs tyrosine phosphorylation and, consequently, mitogenic signaling of the TCR. Reduced MAPK activity-dependent FLICE-like inhibitory protein (FLIP) expression in asmase-/- T lymphocytes increases their sensitivity towards Fas-mediated apoptosis. PMID- 10800076 TI - The fate of the nuclear matrix-associated-region-binding protein SATB1 during apoptosis. AB - Special AT-rich sequence-binding protein 1 (SATB1), predominantly expressed in thymocytes, was identified as a component of the nuclear matrix protein fraction. Programmed cell death of Jurkat T-cells was induced by various stimuli in Fas dependent and -independent fashion. During apoptosis, but not during necrosis, SATB1 was cleaved, as rapidly as was lamin B, in a caspase-dependent way yielding a stable 70 kDa fragment. The same result was obtained for apoptotic HL60-cells. We constructed various deletion constructs of SATB1, expressing protein chimeras tagged with green fluorescent protein (GFP). Transient transfection of these into Jurkat or HeLa cells followed by initiation of apoptosis allowed us to map the potential caspase-6 cleavage site VEMD to the N-terminal third of SATB1, leaving an intact DNA-binding domain in the C-terminal part of the protein. Our results suggest that apoptosis-specific breakdown of SATB1, a transcriptional activator of the CD8a gene, might be of physiological relevance during thymic clonal deletion and apoptosis of peripheral T-lymphoid cells. PMID- 10800077 TI - Different expression patterns of Bcl-2 family genes in breast cancer by estrogen receptor status with special reference to pro-apoptotic Bak gene. AB - Oncogenic and anti-apoptotic Bcl-2 is expressed much less in estrogen receptor alpha (ERalpha) negative breast cancers, which show more malignant phenotypes, than ERalpha-positive, indicating that some other Bcl-2 family member(s) are involved in the apoptotic balance of the cancer cells. We first analyzed mRNA expression of pro-apoptotic Bak and Bax along with that of anti-apoptotic Bcl-2 and Bcl-xL, using breast cancer specimens of 27 patients. Bak mRNA was expressed much less in ERalpha negative breast cancers, along with reduced expression of Bcl-2. Immunostaining of sections of 108 patients confirmed the observation. Next, stable transformants of MCF-7 cells with sense Bak expression vector showed fewer colonies in soft agar compared with the parental cells, while stable introduction of antisense Bak vector enhanced colony formation at lower estradiol concentrations. The reduction of Bak may play important roles in malignant development of breast cancer to acquire estrogen independency, counteracting the reduced Bcl-2. PMID- 10800079 TI - Increased expression of the potential proapoptotic molecule DD2 and increased synthesis of leukotriene B4 during allograft rejection in a marine sponge. AB - Sponges (Porifera) are a classical model to study the events during tissue transplantation. Applying the 'insertion technique' autografts from the marine sponge Geodia cydonium fuse within 5 days. In contrast, allografts are rejected and destroyed. Here we show that during allograft rejection the cells in the grafts undergo apoptosis; 5 days after transplantation 46% of the cells show signs of apoptosis. In a previous study it was shown that during this process a tumor necrosis factor-like molecule is induced in allo- and xenografts. Molecules grouped to the superfamily of tumor necrosis factor receptors and a series of associated adapter molecules contain the characteristic death domain. Therefore, we screened for a cDNA encoding such a domain. Here we report on the first invertebrate molecule from Geodia cydonium comprising a death domain. The potential proapoptotic molecule DD2, with a calculated Mr of 24 970, possesses in contrast to all known mammalian death domain-containing proteins two such domains with highest similarity to the death domain present in human Fas/APO-1. The expression of this gene is not detectable in control tissue but strongly upregulated in allografts; only very low expression is seen in autografts. Parallel with the increase of the expression of the potential proapoptotic molecule DD2 in allografts the level of LTB4 drastically increases from 2.5 pg/mg of protein (controls) to 389 pg LTB4/mg during a period of 5 days after transplantation; the level of LTB4 in autografts does not change. Very likely in response to inflammatory reactions the LTB4 metabolizing enzyme LTB4 12-hydroxy dehydrogenase is expressed both in auto- and allografts. These results demonstrate that sponges are provided with apoptotic pathways, similar to those present in deuterostomes and apparently absent in protostomes, which are composed of molecules comprising a death domain. In addition, it is suggested that in sponges LTB4 is one metabolite which is involved in the initiation of apoptosis. It is postulated that the potential proapoptotic effect of LTB4 is prevented in auto-grafts by the expression of the LTB4 12-hydroxy-dehydrogenase. PMID- 10800078 TI - Retinoid-dependent growth inhibition, differentiation and apoptosis in acute promyelocytic leukemia cells. Expression and activation of caspases. AB - In the NB4 model of acute promyelocytic leukemia (APL), ATRA, 9-cis retinoic acid (9-cis RA), the pan-RAR and RARalpha-selective agonists, TTNPB and AM580, induce growth inhibition, granulocytic differentiation and apoptosis. By contrast, two RXR agonists, a RARbeta agonist and an anti-AP1 retinoid have very limited activity, ATRA- and AM580-dependent effects are completely inhibited by RAR antagonistic blockade, while 9-cis RA-induced cell-growth-inhibition and apoptosis are equally inhibited by RAR and RXR antagonists. ATRA, 9-cis RA and AM580 cause upregulation of the mRNAs coding for pro-caspase-1, -7, -8, and -9, which, however, results in increased synthesis of only pro-caspase-1 and -7 proteins. These phenomena are associated with activation of pro-caspase-6, -7 and -8, cytochrome c release from the mitochondria, inversion of Bcl-2/Bax ratio and degradation of PML-RARalpha. Caspase activation is fundamental for retinoid induced apoptosis, which is suppressed by the caspase-inhibitor z-VAD. PMID- 10800080 TI - Activation of group I metabotropic glutamate receptors reduces neuronal apoptosis but increases necrotic cell death in vitro. AB - Glutamate released during acute CNS insults acts at metabotropic glutamate receptors (mGluR), including group I mGluR. Blockade of group I mGluR during in vitro neuronal trauma provides neuroprotection, whereas activation exacerbates such injury. However, the effects of group I mGluR agonists or antagonists have been primarily studied in in vitro models characterized by necrotic cell death. We examined the role of group I mGluR in the modulation of neuronal injury induced during oxygen-glucose deprivation (OGD), a well-studied model of necrosis, and by application of two well established pro-apoptotic agents: staurosporine and etoposide. Inhibition of group I mGluR attenuated necrosis induced by OGD, whereas selective activation of group I mGluR exacerbated such injury. In contrast, activation of group I mGluR, including selective activation of mGluR5, significantly attenuated apoptotic cell death induced by both staurosporine and etoposide. This effect was completely reversed by co application of a group I mGluR antagonist. Thus, group I mGluR appear to exhibit opposite effects on necrotic and apoptotic neuronal cell death. Our findings suggest that activation of mGluR1 exacerbates neuronal necrosis whereas both mGluR1 and mGluR5 play a role in attenuation of neuronal apoptosis. PMID- 10800081 TI - Properties of DNA fragmentation activity generated by ATP depletion. AB - Internucleosomal DNA fragmentation is generally perceived as one of the characteristic features of apoptosis, most of which are driven by caspase activation dependent upon ATP. On the other hand, ATP depletion has been reported to induce apoptosis accompanying DNA fragmentation. To address this apparent paradox, we analyzed the DNA-fragmenting activity generated in ATP-depleted cells. In HL-60 promyelocytic leukemia cells cultured in glucose-free medium with oligomycin, internucleosomal DNA fragmentation occurred as an early event. The DNA fragmentation was blocked by serine protease inhibitors but not by caspase inhibitors. Consistently, ICAD/DFF45 could not inhibit the DNA-fragmenting activity of the ATP-depleted cytosol in a cell-free system. When ATP was supplied to the cell-free assay, 80% of the DNA-fragmenting activity was lost. The reduced activity was then restored by proteasome inhibitors, suggesting a role of proteasome to protect from a cellular insult derived from ATP-depletion. PMID- 10800082 TI - Conditional expression of anti-apoptotic protein p35 by Cre-mediated DNA recombination in cardiomyocytes from loxP-p35-transgenic mice. AB - p35, a viral inhibitor of caspase, prevents cell death induced by various stimuli. We established an experimental system to study the involvement of caspases in cell death, using primary cultured cells from p35 transgenic mice in which the p35 open reading frame (ORF) had been disrupted by the insertion of a DNA segment flanked by loxP sites, the Cre recognition sites. In this system, p35 expression can be initiated by Cre recombinase. Cardiomyocytes, which are highly sensitive to hypoxic stress, were infected with an adenovirus carrying the cre gene (AxCANCre). Expression of p35 by infection with AxCANCre resulted in inhibition of caspase-3 activation and resistance to hypoxia-induced cell death. Hypoxia-induced cytochrome c release was also attenuated in p35-expressing cardiomyocytes. Our transgenic mice can be used as an experimental model for studying the involvement of caspases in various degenerative diseases as well as programmed cell death both in vitro and in vivo. PMID- 10800083 TI - Molecular characterization of the surface of apoptotic neutrophils: implications for functional downregulation and recognition by phagocytes. AB - We have used a panel of monoclonal antibodies and lectins to examine the profile of surface molecule expression on human neutrophils that have undergone spontaneous apoptosis during in vitro culture. Neutrophil apoptosis was found to be accompanied by down-regulation of the immunoglobulin superfamily members PECAM 1 (CD31), ICAM-3 (CD50), CD66acde, and CD66b and the integrin-associated proteins CD63 and urokinase plasminogen activator receptor (CD87) that may alter the potential for adhesive interactions. Cellular interactions may be further influenced by the reduction of the expression of surface carbohydrate moieties, including sialic acid. Reduced expression of FcgammaRII (CD32), complement receptor type 1 (CD35) and receptors for pro-inflammatory mediators C5a (CD88) and TNFalpha (CD120b) associated with apoptosis might limit neutrophil responsiveness to stimuli that trigger degranulation responses. Although many of the receptors we have examined are expressed at reduced levels on apoptotic neutrophils, we found that there was differential loss of certain receptors (e.g. CD16, CD15 and CD120b) and increased expression of aminopeptidase-N (CD13). Together with our previous data showing that expression of certain molecules e.g. LFA-3 (CD58) is not altered during neutrophil apoptosis, these data are suggestive of specific changes in receptor mobilisation and shedding associated with apoptosis. Although reduced expression of CD63 (azurophilic granules) and CR1 (specific granules) indicates that granule mobilisation does not accompany apoptosis, a monoclonal antibody (BOB78), that recognises a 90 kDa antigen localised in intracellular granules, defines a subpopulation of apoptotic neutrophils that exhibit nuclear degradation yet retain intact plasma membranes. BOB78 positive neutrophils were found to bind biotinylated thrombospondin, suggesting that this mAb defines surface molecular changes associated with exposure of thrombospondin binding moieties. PMID- 10800084 TI - Arguments for the ethical permissibility of transgenic xenografting. PMID- 10800085 TI - Liposome advances: progress in drug and vaccine delivery, Fourth International CDDR Conference, 13-17 December 1999. PMID- 10800086 TI - Neutralisation of adenovirus infectivity by ascitic fluid from ovarian cancer patients. AB - Animal models and phase I clinical trials have shown that repeat virus delivery and subsequent transgene expression is limited by the generation of humoral and cellular immune responses directed towards the therapeutic vector. The presence of a pre-existing immune response may even prevent initial delivery. In order to determine the presence of pre-existing anti-adenovirus humoral immunity we analysed ascitic fluid, collected from the peritoneal cavity of patients with advanced ovarian cancer. Twelve ascitic fluid and four matched serum samples were examined. The titre and isotype of anti-adenovirus antibodies was determined by ELISA, and Western blotting identified the molecular basis of the immune response, which was primarily directed towards fibre and penton base. Neutralisation of virus infectivity was assessed in vitro by measurement of green fluorescent protein reporter gene expression. We found that the ascitic fluid samples contain antibodies that recognise both adenovirus types 2 and 5, were predominantly IgG and directed towards the viral antigens responsible for cell adhesion, and had virus neutralising activity. PMID- 10800087 TI - Selective in vivo transfection of murine biliary epithelia using polycation enhanced adenovirus. AB - We have investigated the use of polycations to increase adenovirus-mediated expression of transgenic protein to the biliary epithelia with a view to gene therapy for hepatobiliary disease in CF. We have shown that adenovirus carrying the beta-galactosidase transgene transfect both human and mouse biliary epithelia in primary culture and that in both instances adenovirus transfection can be significantly increased by co-complexing with polycation. In vivo administration of 1 x 109 p.f.u. adenovirus co-complexed with the polyamine polyethyenimine (PEI) into the mouse biliary duct leads to >80% positively stained biliary epithelia while adenovirus alone at the same titre infected <5% biliary epithelia. We suggest that the use of low titre polycation enhanced adenoviral delivery to the biliary tree of CF patients could be of therapeutic significance. As a prelude to an extensive in vivo functional investigation in CF null mice we have shown that Ad5/polycation complexes deliver functional CFTR to non-CFTR expressing cells in vitro more efficiently than Ad5 alone. PMID- 10800088 TI - Long-term silencing of retroviral vectors is resistant to reversal by trichostatin A and 5-azacytidine. AB - One problem limiting the development of long-term gene replacement therapy is gene silencing. A variety of experiments have implicated DNA methylation and histone deacetylation in gene silencing and shown that the agents 5-azacytidine (5-Aza) and trichostatin A (TSA) are able to reverse these effects. To begin to investigate clinically relevant strategies to reverse silencing with these drugs, we transduced the MEL and FDCP-1 hematopoietic cell lines with Moloney murine leukemia virus (MMLV) and Harvey murine sarcoma virus (HMSV)-based retroviral vectors carrying the beta-galactosidase/neomycin resistance fusion gene (beta geo). Fifty-one clones were isolated under G418 selection over 2 weeks and then allowed to grow without selection as beta-gal activity was monitored over time. More than 80% of these clones showed significant silencing over a period of 70-80 days. The clones were then exposed to a wide range of 5-Aza and TSA concentrations, both alone and in combination, in an effort to reverse silencing. Despite demonstration that the agents were able to decrease DNA methylation and increase histone acetylation, significant reversal of long-term silencing was not seen under any experimental condition. These results suggest that long-term retroviral silencing involves mechanisms in addition to DNA methylation and histone acetylation and that new pharmacologic strategies are needed to overcome the silencing process. PMID- 10800089 TI - Functional long-term thymidine kinase suicide gene expression in human T cells using a herpesvirus saimiri vector. AB - Herpesvirus saimiri transforms human T lymphocytes to stable growth and persists episomally without genomic integration and without virus production. The transformed T cells retain essential features of their parental cells including the MHC-restricted antigen specificity which may be useful for applications in adoptive immunotherapy. In order to improve the biological safety of such vectors, the prodrug activating gene thymidine kinase of herpes simplex virus was inserted into the genome of herpesvirus saimiri by homologous recombination. After infection with wild-type or cloned recombinant viruses, T cells from tamarin monkeys and from humans were transformed to stable growth. Thymidine kinase-expressing transformed T cells were efficiently eliminated in the presence of low concentrations of ganciclovir. This elimination mechanism remained fully functional over an observation period of 12 months. The potentially immunogenic neomycin resistance gene expression cassette was deleted from the genome of established mutant viruses by using the prokaryotic Cre/LoxP recombination system. At any time during the course of a therapeutic application, thymidine kinase-expressing transformed human T cells might be eliminated after administration of ganciclovir. In principle, this function could be useful for the T cell-dependent immunotherapy of resistant blood cancer while avoiding the risk of uncontrolled graft-versus-host disease. PMID- 10800090 TI - Gene transfer to adult human lung tissue ex vivo. AB - The potential of gene therapy for treatment of lung disease remains unrealised. Early model systems often resulted in promising efficiency of gene transfer, only to prove irreproducible in the clinic. While problems such as induction of host immune responses and duration of expression also need to be addressed, it is now widely believed that alternative, relevant models which more accurately reflect gene transfer efficiencies in human lungs are urgently required. We report here on a human lung slice culture system to assess gene transfer to adult lung epithelium. A lacZ-expressing adenovirus (AdCA35lacZ) was used as a reporter vector. A solution of AdCA35lacZ was instilled via bronchioles into resected lung tissue, a route analogous to clinical administration. Following a 1 h incubation, the tissue was inflated with a 0.4% agarose solution, instilled via the same bronchioles. Once solidified, 500 microm slices of the tissue were prepared and cultured for 4 days. beta-Galactosidase staining revealed lacZ transgene expression in bronchiolar and alveolar cells of the lung slices throughout the 4 days in culture. This system, which can also be used to study other viral and liposome vectors, could prove to be a useful alternative model for assessing gene delivery to adult human lung epithelium. PMID- 10800091 TI - Central nervous system toxicity of two adenoviral vectors encoding variants of the herpes simplex virus type 1 thymidine kinase: reduced cytotoxicity of a truncated HSV1-TK. AB - Herpes simplex virus type 1-thymidine kinase (HSV1-TK) in combination with ganciclovir is an efficient and widely used strategy in brain tumour gene therapy. Recently, we have shown effective inhibition of glioma growth in a syngeneic rat model using recombinant adenoviruses expressing the full-length HSV1-TK and an N-terminus truncated variant, HSV1-DeltaTK in the presence of ganciclovir. We also showed active chronic brain inflammation in the long-term survivors (3 months) treated with HSV1-TK plus GCV. Furthermore, our results indicated loss of myelinated fibres, oedema and indices of ongoing axonal degeneration. In this study, we assessed the cytotoxicity of both HSV1-TK variants in the presence or absence of ganciclovir, in primary cultures of neurones and glia, and in the rat brain in vivo. Our results indicate that, at viral doses where tumour cells are sensitive to the enzyme/prodrug system, (1) there is no major cytotoxicity for either neurones or glial cells grown in primary cultures, (2) on its own the full-length HSV1-TK is more cytotoxic than its truncated version HSV1-DeltaTK for a population of non-neuronal and non-glial cells within neocortical primary cultures, and (3) in vivo, when delivered into the striatum, RAds encoding HSV1-TK are more cytotoxic than RAds encoding HSV1 DeltaTK, after administration of ganciclovir. The effectiveness of HSV1-DeltaTK in preventing brain tumour growth in vivo, combined with its reduced cytotoxicity, both in vivo and in primary cultures of CNS cells, could represent an advantage for treatment of brain tumours using gene therapy. PMID- 10800092 TI - Glioma/glioblastoma-specific adenoviral gene expression using the nestin gene regulator. AB - For glioma- and glioblastoma-specific gene expression, we utilized a nestin regulatory element whose activity was evaluated by the reporter gene lacZ. Nestin is a 38-kDa intermediate filament protein, and is expressed specifically in the neuroepithelial stem cells. Nestin is detected in gliomas and glioblastomas, but not in normal brain tissue. We constructed a nestin gene regulator by placing nestin's second intron before the 5' upstream region (2iNP). To obtain enhanced expression of this tissue-specific regulator, we utilized the adenovirus double infection method with a Cre-loxP on/off switching system. We constructed a 'regulator' vector, Ax2iNPNCre, which expresses Cre recombinase under the control of the nestin regulatory element, 2iNP. A 'reporter' vector, AxCALNLNZK, expresses lacZ under the control of a strong CAG promoter when the stuffer sequence has been removed by Cre recombinase at a pair of loxP sites. We used seven human glioma/glioblastoma cell lines: U251, KG-1C, NGM5, U87 MG, LN-Z308, NP-2 and T98G. Of these, nestin was expressed highly in U251 and KG-1C, less in NGM5, and undetectably in the other four lines. With the use of the two adenovirus vectors, we found X-gal staining and high nestin regulator-promoted beta-galactosidase activities in four of the seven glioma/glioblastoma cell lines. Staining was strong in U251, KG-1C and NGM5, and less in U87 MG. LacZ expression was nearly undetectable in the non-glioma cell line, HeLa, but a little in COS-7. The adenovirus double-infection method, which uses a nestin regulator, is applicable for glioma/glioblastoma-specific expression. PMID- 10800093 TI - 8 Br-cAMP enhances both humoral and cell-mediated immune responses induced by an HIV-1 DNA vaccine. AB - From a series of preclinical studies and animal experiments, we have been able to demonstrate that DNA vaccines are a promising tool in strategies for protecting hosts from a variety of infectious diseases. Since the promoter activity of the human cytomegalovirus immediate-early promoter/ enhancer (CMV promoter) is known to be responsive to an elevation in the level of intracellular cAMP, we hypothesized that use of cAMP analogue (8-Bromo adenosine 3'5'-cyclic monophosphate, 8 Br-cAMP) would increase the level of transgene expression supported by the CMV, and enhance the ability of DNA vaccines to evoke an immune response against the transgene product in vivo. To evaluate this hypothesis, immune responses against HIV-1 envelope protein, gp160, an immunogenic HIV-1 component expressed under the control of the CMV promoter, were evaluated in BALB/c mice with or without stimulation by 8 Br-cAMP. DNA vaccine with 8 Br-cAMP was intramuscularly (i.m.) or intranasally (i.n.) administered to BALB/c mice twice on days 0 and 14. Regardless of which route was used, the combination increased the serum IgG antibody (Ab) titer, HIV-1-specific cytotoxic T lymphocyte (CTL) activity and the delayed-type hypersensitivity (DTH) response, compared with the effect of using the vaccine alone. When administered via the i.n. route, the combination also remarkably increased the titer of secretory IgA (sIgA). Moreover, it induced increased production of interferon-gamma with reduction in IL-4 synthesis, and decreased the ratio of serum IgG1/IgG2a. However, these enhancements were not observed when 8 Br-cAMP was coadministered with peptide vaccine or protein antigen. These data suggest that 8 Br-cAMP is able to enhance both humoral and cellular immune responses induced by the DNA vaccine. The induction of T helper type 1 (Th1) immunity against HIV-1 was also enhanced by coadministration of 8 Br-cAMP. A CAT assay study demonstrated that the adjuvant effect of 8 Br-cAMP may be due to the activation of the CMV promoter in the DNA vaccine. The virus challenge experiment in a mouse influenza model also proved our hypothesis. PMID- 10800094 TI - DNA vaccination with full-length or truncated neu induces protective immunity against the development of spontaneous mammary tumors in HER-2/neu transgenic mice. AB - Genetic immunization against tumor antigens is an effective way to induce an immune response able to oppose cancer progression. Overexpression of HER-2/neu can lead to neoplastic transformation and has been found in many human primary breast cancers. We constructed DNA expression vectors encoding the full-length neu oncogene of rat cDNA (pCMV-NeuNT), the neu extracellular domain (pCMV-ECD), or the neu extracellular and transmembrane domains (pCMV-ECD-TM). We evaluated whether i.m. injection of these plasmids induces protection against the development of mammary tumors occurring spontaneously in FVB/N neu-transgenic mice. We found that pCMV-ECD-TM induced the best protection, whereas both pCMV ECD and pCMV-NeuNT were less effective. The coinjection with a bicistronic vector for murine IL-12 increased the efficacy of pCMV-ECD and pCMV-NeuNT plasmids, and led to the same protection obtained with pCMV-ECD-TM alone. Anti-neuECD antibodies were detected in pCMV-ECD-TM vaccinated mice and, after coinjection with pCMV-IL12 plasmids, they appeared also in animals immunized with pCMV-ECD. Our data demonstrate the effectiveness of DNA vaccination using truncated Neu plasmids in inducing antitumor protection in a spontaneous mammary tumor model. PMID- 10800095 TI - Induction of potent antitumor response by vaccination with tumor lysate-pulsed macrophages engineered to secrete macrophage colony-stimulating factor and interferon-gamma. AB - Adoptive transfer of activated macrophages, being both effector cells and antigen presenting cells, represents a promising approach to immunotherapy of cancer. In order to get activated macrophages with increased antitumor potential, in the present study, murine peritoneal macrophages were transduced with human macrophage colony-stimulating factor (M-CSF) and murine interferon-gamma (IFNgamma) by recombinant adenovirus infection. The results demonstrate that M CSF and IFNgamma gene-modified macrophages exhibited higher expression of MHC-II, B7.1 and ICAM-1, increased antigen-presenting activity and cytotoxicity. It was also shown that they secreted more tumor necrosis factor, interleukin-1 and nitric oxide. In vivo experiments showed that in previously initiated murine pulmonary metastatic melanoma, tumor lysate-pulsed, M-CSF and IFNgamma gene modified macrophages elicited more potent antitumor effects than tumor lysate pulsed M-CSF or IFNgamma gene-modified macrophages. Cytotoxic T lymphocyte (CTL) activity, IFNgamma and tumor-necrosis factor production of the splenocytes increased significantly in mice after intravenous injection of the gene-modified macrophages. M-CSF and IFNgamma gene-modified macrophages may act as activated effector and antigen-presenting cells, thus eliciting a more potent antitumor response. PMID- 10800146 TI - Racing ahead. PMID- 10800147 TI - Community paediatrics moves on-an analysis of changing work patterns 1994-97. AB - Objectives: To describe the current clinical workload of the modern community paediatrician; to outline the changes in this role over recent years and examine the reasons for these. Design and setting: The design is a retrospective analysis of data routinely collected for contracting purposes. Nottingham community paediatric service 1994-97 is the setting. Main outcome measures: These are the characteristics of patients seen, sources of referral, locations of clinical contact, referral rates by area. Results: 36 710 appointments were offered over the time period studied. The non-attendance rate was 17%. Pre-school children made up the largest group seen. Most referrals were from health visitors (23%) and school nurses (29%). There has been a small but significant increase in the numbers of children seen who have developmental problems and disability, and due to child protection issues between the time periods. There has been a shift in the proportions of children seen in a local health centre referral clinic (9.8% increase during 1994-97) rather than in a school setting. Twice as many children are referred to the community paediatrician from inner city areas than from the surrounding county areas. Conclusion: The transfer of child health surveillance to the primary health care team and the increased training of community paediatricians, has resulted in community paediatricians developing an increasingly specialised role particularly in the areas of child growth and development, disability, social and behavioural paediatrics. Public Health (2000) 114, 61-64. PMID- 10800096 TI - Engineering mouse T lymphocytes specific to type II collagen by transduction with a chimeric receptor consisting of a single chain Fv and TCR zeta. AB - The chimeric cell surface receptor scC2Fv/CD8/zeta was constructed to engineer primary mouse T lymphocytes with antibody-type specificity to type II collagen (CII). Such cells could be used as gene carriers in the anti-inflammatory gene therapy of an autoimmune arthritis. This receptor includes the single chain Fv domain (scFv) of the anti-CII monoclonal antibody (mAb) C2, hinge region of CD8alpha and the transmembrane and cytoplasmic domains of TCRzeta. The scC2Fv/CD8/zeta gene was transduced into T cell hybridomas and primary mouse lymphocytes using retrovirus-mediated gene transfer. The chimeric receptor scC2Fv/CD8/zeta forms covalently bound homodimers, as demonstrated in T cell hybridomas and packaging fibroblasts. It does not associate with endogenous signalling subunits of the TCR complex. When scC2Fv/CD8/zeta-expressing clones of T cell hybridomas MD.45 and HCQ6 were stimulated with CII they produced IL-2. The level of their IL-2 response correlated with the expression level of the chimeric receptor on the cell surface. Splenocytes isolated from DBA/1 mice were stimulated with Con A in vitro to facilitate retrovirus-mediated transfer of the scC2Fv/CD8/zeta gene. As a result of transduction, approximately 4% of the Con A activated splenocytes expressed the chimeric receptor scC2Fv/CD8/zeta on the cell surface. These cells proliferated in response to stimulation with CII. PMID- 10800148 TI - The society of public health: 1957-1997. AB - History of the Period 1957-1997 The Society of Public Health: 1957-1997 is in two parts that are complementary but may be read independently of each other. Part 1: A History of the Period 1957-1997 This account details the progress and activity of the last forty years of the Society of Public Health (previously The Society of Community Medicine: Originally The Society of Medical Officers of Health) before its ammalgamation with the Royal Institute of Public Health and Hygiene to form The Royal Institute of Public Health and Hygiene and Society of Public Health. The appendices to this account cover the Presidents of The Society and Its Branches (Home And Overseas) and Special Interest Groups, Editors, its Faculty, Awards and Conferences since the Society's establishment in 1856 or their own beginning, as appropriate. Published in Public Health (2000) 114, 73 107 Part 2: Profiles of Some Fellows of the Society of Public Health To be published in Public Health (2000) 114, (3) This consists of profiles of eminent people active within the Society and the wider discipline of public health during the period 1957-1997. As it is based on availability of information the series cannot be all inclusive and readers are invited to consult the history in Part 1 and the listings in its appendices for additional information regarding members of the Society of Public Health. PMID- 10800149 TI - The effects of dietary supplementation on growth and adult mortality: a re analysis and follow-up of a pre-war study. AB - The aim of this study was to re-analyse a diet supplementation study conducted in the 1930s and investigate the effects of food supplementation on children's growth and later adult mortality. A non-randomised controlled trial was carried out in eight of the sixteen centres participating in the Carnegie Survey of Diet and Health in pre-war Britain (1937-39). Food supplements were given for 12 months either at school or as food parcels sent to the family home. 545 children aged 2-14 received food supplements and 494 children of a similar age acted as their controls. The children came from 465 families. The increase in childhood height and its components-leg length and trunk length-over 12 months follow-up were measured. Mortality from all causes over 60 years follow-up to 1998 was also assessed. There were important differences between fed and control children at baseline. Supplemented children came from larger families with poorer diets and most were examined, on average, 12 days later than control children. After adjustment for baseline imbalances those receiving supplements increased in height by 3.7 mm (95% CI 1.9-5.5) more than the controls. After adjustment, most of the difference in growth appeared to arise as a result of increases in leg length. After adjustment for confounding factors measured in childhood, no significant effect of childhood food supplements on adult mortality was seen. The age-adjusted hazard ratio for all cause mortality was higher in the supplemented compared to control subjects: 1.13 95% CI (0.77-1.64). We found that provision of childhood food supplements led to increased growth amongst supplemented children. The increases in height in this study were mainly as a result of increases in leg length and are similar to those found in a more recent randomised trial in South Wales. Whilst other analyses suggest that childhood height is important in predicting adult mortality patterns, we found no significant effect of childhood food supplements on adult mortality patterns in this study, although the study lacked statistical power to detect small but, nevertheless, important differences in mortality. Larger randomised trials with long term follow-up would be required to investigate the impact of childhood food supplementation on adult health. PMID- 10800150 TI - The influence of gender, social circumstances and smoking on survival: the Renfrew and Paisley study. AB - The present study aimed to compare survival to age 75 between men and women, by social circumstances and smoking behaviour. A 20 year follow up was carried out of a large representative cohort of Scottish men and women in the Renfrew and Paisley study, and was combined with Scottish mortality statistics. 6831 men and 7993 women aged 45-64 y at time of examination (between 1972 and 1976) were considered. Combining the estimates from the Renfrew and Paisley study with those from Scottish mortality statistics for men and women younger than 45 y of age, it was calculated that 28% of the male smokers and 44% of the male never smokers in disadvantaged social circumstances will have survived to age 75 y, compared with 46% and 56% of the female smokers and never smokers respectively. In more privileged social circumstances, 41% of the male smokers and 62% of the male never smokers will have survived to age 75 y, compared with 56% and 70% of the female smokers and never smokers respectively. The difference between male smokers in low social classes and female never smokers in high social classes is 42% in absolute terms (28% vs 70%), which gives an indication of the combined influence of gender, social circumstances and smoking on survival. These results show that gender, social circumstances and smoking are important determinants of mortality which have led to substantial differences in survival. The influence of additional factors related to mortality could usefully be compared to these to put their effects into perspective. PMID- 10800151 TI - Bidi smoking and health. AB - Bidi, made of low-grade tobacco, is popular among rural folk and the urban poor, in the South Asian region. The objectives of this study were: (1) to assess the extent of bidi smoking in different countries; (2) to compare its smoke and health hazards with conventional cigarettes and; (3) to recommend preventive measures. English language articles were identified through a MEDLINE computerised search (1966-1998) using the term 'bidi', 'beedi', 'beedie', 'biri' and manually in order to collate all the available information and to present a descriptive epidemiological profile. The prevalence of bidi smoking was found to be 21-56% among men in South Asian countries with very little information available about women. Concentrations of nicotine, tar and other toxic agents in the smoke are higher for bidi than for other cigarettes. Bidi smoking is also considered to cause about 2-3 times greater nicotine and tar inhalation than do conventional cigarettes, due to the poor combustibility of the bidi wrapper and greater puff frequency needed to keep the bidi alight. It has also been shown to have greater odds ratios, although not statistically significant, for various cancerous and chronic conditions in comparison with other cigarettes. Since bidi is hand-rolled, workers employed in bidi factories are at risk of developing cancerous conditions due to exposure to tobacco dust and flakes. We suggest that anti-smoking campaigns should be directed to the bidi smokers of all South Asian countries. Most bidi smokers are illiterate and malnourished which makes them more vulnerable to smoking-related morbidity and mortality. At the same time, measures should be taken to make bidi less harmful and to produce bidi by machine to reduce exposure-related toxicity among bidi production workers. PMID- 10800152 TI - Do military dentists advise tobacco users to quit? The patient's perspective. AB - This study explores whether tobacco users in the US military report having been counseled against tobacco use by a military dentist. The data come from a 26-site survey of active duty Army, Navy, Marine, and Air Force personnel conducted from April 1994 to January 1995. A prestratified, random sample was drawn. Women and minorities were oversampled. Respondents provided demographic and tobacco use data by self-administered questionnaires. Of 12 950 respondents (81% response rate), 4777 reported using some type of tobacco. Prior to analysis, the data were weighted to reflect the military population. Bivariate and logistic regression analysis were employed to determine what demographic and clinical factors are associated with having been counseled against tobacco use. Results show that among tobacco users, cigarettes (73%) are the most popular type of tobacco consumed followed by snuff (23%), pipes (12%), chewing tobacco (8%), and cigars (5%). Advisement to cease tobacco use varies across type of tobacco consumed, with snuff users (72%) most likely and pipe smokers (57%) least likely being advised to quit. Logistic regression results show that advisement to quit tobacco use varies across patient demographics but is unrelated to periodontal health status. Military dentists should be encouraged to counsel all patients who use tobacco to quit. PMID- 10800153 TI - Potential of using existing injury information for injury surveillance at the local level in developing countries: experiences from Bangladesh. AB - Although injury is a major public health problem in Bangladesh, as in other low income countries in the world, it has gained very little attention from policy makers, or even health professionals, mainly due to the lack of valid injury information in these countries. To address the injury problem, there is an urgent need to develop injury surveillance activities, based on existing sources of potential interest for obtaining injury information. This study is an evaluation of existing injury data sources in terms of prevention orientation representativeness, and sustainability with respect to the use of these sources in injury surveillance at the local level in Bangladesh. The regular injury information sources that have been used in this study include hospital records, post-mortem reports, newspaper reports, and police reports. In addition, 60 health practitioners in the area, representing various categories, were interviewed according to questionnaire, and ten others were interviewed in-depth about their opinions regarding their participation in the injury surveillance system. The information sources were then assessed for their potential to be included in the surveillance system. The validity was estimated by means of comparisons of the different sources. All the existing sources of injury information suffer from certain deficiencies such as, lack of representativeness, low prevention orientation and poor sustainability. Post-mortem reports and police reports have been found to be potential sources of mortality data but not morbidity data. Hospital records can be seen as potential sustainable sources of injury morbidity data but suffer from low prevention orientation and representativeness. The injury information was not consistent from source to source and failed to depict the true picture of injury in the area. To develop an injury surveillance system, a combination of data obtained from several different sources is necessary to compensate for inadequacies in the individual sources and to improve the overall usefulness of the system. Updated with E-codes, hospital discharge records could be used for injury morbidity surveillance, and updated police and post-mortem data can be used for mortality data in injury surveillance systems at the local level in developing countries. PMID- 10800154 TI - The agreement between self-reporting and clinical diagnosis for selected medical conditions among the elderly in Taiwan. AB - A cross-sectional survey was conducted to consider whether there is agreement between self-report and clinical diagnosis in the prevalence of selected common chronic diseases among elderly Taiwanese. Between July 1992 and June 1993, both face-to-face interview and clinical evaluation were applied to a sample of 228 Taiwanese aged 65 y and older to estimate the prevalence of heart diseases, hypertension and diabetes. The results showed that a self-reported history of diabetes had the highest sensitivity (66. 7%) and specificity (95.2%). The self report of heart diseases was the least sensitive (20.5%), while self-reports of hypertension was the least specific (82.8%). Multivariate analyses showed that age, education and number of self-reported diseases suffered from, appeared to influence the accuracy of the self reported data. Our data suggest a notable lack of agreement abetween self-report and clinical diagnosis for medical conditions, even for those with clear and unambiguous diagnostic criteria. The elderly in Taiwan tended to under report (with a consistently higher false negative rate than false positive rate) the existence of the three selected medical conditions. This was especially so for heart diseases. Our findings suggest that, if the level of disease prevalence in the population needs to be known for health planning estimates for hypertension and diabetes would not necessarily be a problem. However, the prevalence for heart disease would be seriously under estimated. We also argue that information obtained by self-report should be validated before use, especially in the case of assessment of the prevalence of medical conditions in the elderly. PMID- 10800155 TI - Breast cancer in Iran: a review of 903 case records. AB - A retrospective study was carried out in five main General District and Teaching Hospitals in Tehran (18 general surgical wards) to investigate breast cancer in Tehran, Iran. All patients who have had breast pathology during 1985-95 were reviewed. For each patient the following data were extracted from case records sex, age, type of disease (based on ICD-10 and ICD-O), breast cancer pathology, tumor size, lymph node involvement, pathological staging (based on TNM system), type of surgery, mass location and the affected side of breast. In all 3085 records were reviewed. Of these, in 903 cases the final diagnosis was breast cancer. The mean age of breast cancer patients was 47.1 (SD=12.3) y and the age adjusted analysis showed that the observed number of breast cancer cases in the age group 40-49 was higher than expected (P<0.001). The majority of patients (71%) were patients with infiltrating ductal carcinoma, 70% had advanced disease (Stage III), and in 51% of cases the tumour was more than 5 cm in size. Finally it was found that two-third of patients (76%) underwent modified radical mastectomy. Although this study does not address true incidence, but rather frequency, the findings suggest that the Iranian breast cancer patients are relatively younger than their western counterparts and mostly have locally advanced disease. In terms of breast cancer screening programs, these findings have some implications for public health professionals in Iran. PMID- 10800156 TI - The clinical utility of epoetin in cancer patients: a matter of perspective. PMID- 10800157 TI - Gene therapy for X-linked chronic granulomatous disease. PMID- 10800158 TI - The expression of full length Gp91-phox protein is associated with reduced amphotropic retroviral production. AB - BACKGROUND AND OBJECTIVE: As a single gene defect in mature bone marrow cells, chronic granulomatous disease (X-CGD) represents a disorder which may be amenable to gene therapy by the transfer of the missing subunit into hemopoietic stem cells. In the majority of cases lack of Gp91-phox causes the disease. So far, studies involving transfer of Gp91-phox cDNA, including a phase I clinical trial, have yielded disappointing results. Most often, low titers of virus have been reported. In the present study we investigated the possible reasons for low titer amphotropic viral production. DESIGN AND METHODS: To investigate the effect of Gp91 cDNA on the efficiency of retroviral production from the packaging cell line, GP+envAm12, we constructed vectors containing either the native cDNA, truncated versions of the cDNA or a mutated form (LATG) in which the natural translational start codon was changed to a stop codon. Following derivation of clonal packaging cell lines, these were assessed for viral titer by RNA slot blot and analyzed by non-parametrical statistical analysis (Whitney-Mann U-test). RESULTS: An improvement in viral titer of just over two-fold was found in packaging cells containing the start-codon mutant of Gp91 and no evidence of truncated viral RNA was seen in these cells. Further analysis revealed the presence of rearranged forms of the provirus in Gp91-expressing cells, and the production of truncated, unpackaged viral RNA. Protein analysis revealed that LATG-transduced cells did not express full-length Gp91-phox, whereas those containing the wild-type cDNA did. However, a truncated protein was seen in ATG transduced cells which was also present in wild type cells. No evidence for the presence of a negative transcriptional regulatory element was found from studies with the deletion mutants. INTERPRETATION AND CONCLUSIONS: A statistically significant effect of protein production on the production of virus from Gp91 expressing cells was found. Our data point to a need to restrict expression of the Gp91-phox protein and its derivatives in order to enhance retroviral production and suggest that improvements in current vectors for CGD gene therapy may need to include controlled, directed expression only in mature neutrophils. PMID- 10800159 TI - Human herpesvirus 6 in hematologic diseases in China. AB - BACKGROUND AND OBJECTIVE: The prevalence and pathogenic role of human herpesvirus 6 (HHV-6) in various benign and malignant hematologic diseases remain largely unknown. The aim of this study was to search for a possible involvement of HHV-6 in the pathogenesis of hematologic diseases. DESIGN AND METHODS: The presence of HHV-6 DNA sequences was examined by polymerase chain reaction (PCR) in bone marrow mononuclear cells from 241 patients with benign and malignant hematologic diseases in China. Platelet-associated immunoglobulin (PAIg) of 66 idiopathic thrombocytopenic purpura (ITP) patients was measured by competitive enzyme-linked immunosorbent assay. The presence of HHV-6 DNA in sera from 31 ITP patients was examined by PCR. Paired serum samples from 19 ITP patients were analyzed for anti HHV-6 IgG titers using an indirect immunofluorescence assay. RESULTS: HHV-6 DNA was detected in 41% and 37.5% of ITP and acute leukemia patients respectively, but in only 6.7% of patients with iron deficiency anemia. HHV-6 positivity for ITP patients with excessive PAIgG was significantly higher than in patients with a normal level of PAIgG. HHV-6 DNA was not detected in any of the serum samples from ITP patients. None of the 19 cases of ITP showed a significant increase in anti-HHV-6 antibody titers during the convalescent phase compared with the onset phase. INTERPRETATION AND CONCLUSIONS: Our results indicate that HHV-6 infection might be associated with excessive PAIgG in some cases of ITP, and that the virus persists in a latent state. The pathogenic role of HHV-6 in ITP needs to be confirmed by further investigations. PMID- 10800160 TI - Role of hepatocyte growth factor in the development of dendritic cells from CD34+ bone marrow cells. AB - BACKGROUND AND OBJECTIVE: Hepatocyte growth factor (HGF) is known to augment the effects of stem cell factor, interleukin-3, granulocyte-macrophage colony stimulating factor (GM-CSF), erythropoetin, and granulocyte colony-stimulating factor, all of which are involved in hematopoiesis. HGF is also known to have a role in immune responses. The aim of this study was to investigate whether HGF is involved in the development of dendritic cells (DC) from CD34+ bone marrow cells. DESIGN AND METHODS: CD34+ cells obtained from three healthy donors were incubated in various combinations of HGF, GM-CSF, and tumor necrosis factor (TNF) for 12 days. Developing cell populations were analyzed for surface markers, morphology and functional capacities by flow cytometry, light microscopy and mixed lymphocyte reaction, respectively. RESULTS: Incubation with HGF alone generated greater number of dendritic cells from CD34+ bone marrow cells than incubation with GM-CSF, or a combination of GM-CSF with TNF. HGF was also found to potentiate the effect of GM-CSF on DC and monocyte development. The effects of HGF were inhibited by the concurrent use of TNF. INTERPRETATION AND CONCLUSIONS: HGF appears to be a significant factor in the development of dendritic cells from CD34+ bone marrow cells. PMID- 10800161 TI - Geographic distribution of CDA-II: did a founder effect operate in Southern Italy? AB - BACKGROUND AND OBJECTIVE: Congenital dyserythropoietic anemia type II (CDA-II) is an autosomal recessive condition, whose manifestations range from mild to moderate. Its exact prevalence is unknown. Based on a recently established International Registry of CDA-II (64 unrelated kindreds), a high frequency of CDA II families living in South Italy became evident. DESIGN AND METHODS: The aim of this study was to define the haplotypes of the CDA II kindreds living in Southern Italy based on markers D20S884, D20S863, RPN, D20S841 and D20S908. These markers map to 20q11.2, within the interval of the CDAN2 gene that is responsible for CDA II. Next, we looked at these markers in kindreds from other regions of Italy and from other countries, with special attention to families having ancestors in Southern Italy. RESULTS: Evaluation of the geographic distribution of the ancestry of Italian CDA-II patients clearly demonstrated the unusually high incidence of this condition in Southern Italy. Our statistical calculations and linkage disequilibrium data also clearly demonstrate a strong association of the markers of chromosome 20 with the disease locus in our sample. Almost all the regions defined by the markers here used is in disequilibrium with the disease. Combining the data from the Italian sample together with those obtained from the non-Italian ones, we can restrict the area of highest disequilibrium to that defined by markers D20S863-D20S908. INTERPRETATION AND CONCLUSIONS: Despite the presence of this linkage disequilibrium the search for a common haplotype failed. This could suggest that the mutation was very old or that it occurred more than once on different genetic backgrounds. PMID- 10800162 TI - Differential expression of cellular prion protein on human blood and tonsil lymphocytes. AB - BACKGROUND AND OBJECTIVE: The expression of cellular prion protein (PrPc) on the surface of peripheral lymphocytes has been previously reported, but little is known about its expression on lymphoid cells from secondary lymph organs. In this report, we compare the surface expression of PrPc on human blood lymphocytes and tonsil lymphocytes. DESIGN AND METHODS: This analysis was performed by cytometry on live lymphocytes isolated from healthy donors or from the tonsils of adults or children. RESULTS: Human peripheral lymphocytes and tonsillar lymphoid cells, but not erythrocytes or granulocytes, express PrPc at their surfaces. Interestingly, we found significantly less PrPc on freshly isolated tonsil lymphocytes, both B and T, than on blood cells. Although tonsil cells bear less PrPc than circulating blood lymphocytes, they are able to express high quantities of PrPc on their surface when placed in culture. However, contrary to previous results, mitogen stimulation does not affect this expression on B- or T-cells. INTERPRETATION AND CONCLUSIONS: We suggest that the PrPc expression by lymphocytes may be modified by interactions occurring during intratissular migration or during cell-to-cell contacts. Whether PrPc plays a role in intracellular communication at this location, as it does in the nervous system, remains an open question. PMID- 10800163 TI - Detection of inv(16) and t(16;16) by fluorescence in situ hybridization in acute myeloid leukemia M4Eo. AB - BACKGROUND AND OBJECTIVE: It has been established that cytogenetic findings at the time of diagnosis of acute myeloid leukemia (AML) are powerful prognostic indicators. Pericentric inversion of chromosome 16 and translocation t(16;16) resulting in chimeric fusion of CBFB and MYH11 genes are typically seen in the M4 Eo FAB classification subset of AML and are associated with low-risk disease. These subtle chromosomal abnormalities may be difficult to detect in poor-quality metaphase preparations and if missed could lead to incorrect assignment to risk groups and influence the therapy decision-making process. DESIGN AND METHODS: We prospectively studied, at diagnosis, 10 patients with AML-M4 Eo by cytogenetics and fluorescent in situ hybridization (FISH) with two cosmids (36 and 40). As a control group, 7 patients (5 with a diagnosis of AML other than M4 Eo and two cases of reactive eosinophilia) were analyzed. In addition reverse transcriptase chain reaction (RT-PCR) studies were carried out in 6 cases. RESULTS: Karyotypic analysis detected the inv(16) in all but one of the patients with M4-Eo while none of the control cases showed any abnormality on chromosome 16. FISH studies showed that all 10 patients had abnormalities on chromosome 16; the patient with normal karyotype showed an inv(16) by FISH, while a case with inv(16) by cytogenetics had a t(16;16) by FISH. RT-PCR demonstrated amplification of the CBFB/MYH11 product in all cases analyzed. INTERPRETATION AND CONCLUSIONS: In patients with M4Eo and rearrangements of chromosome 16, FISH studies may afford more complete information than conventional cytogenetics and can be an alternative to RT-PCR studies. PMID- 10800164 TI - Cytogenetic and molecular cytogenetic characterization of 6 new cases of idiopathic hypereosinophilic syndrome. AB - BACKGROUND AND OBJECTIVE: Idiopathic hypereosinophilic syndrome (HES) is defined as a peripheral blood eosiniphilia greater than 1, 500 cells/microL for longer than 6 months, absence of other apparent etiologies for eosinophilia and signs and symptoms of organ involvement. HES may be a reactive condition or a chronic myeloproliferative disorder but scanty information is available concerning its cytogenetic profile. DESIGN AND METHODS: Six patients with HES were studied by cytogenetic analysis. To increase the sensitivity of cytogenetic analysis, interphase FISH studies were performed to detect some cryptic chromosomal lesions involving the regions known to be frequently involved in myeloproliferative disorders (i.e. BCR/ABL, 5q31, 7q31.1, 11q23, 13q14, 17p13). Clinical parameters were recorded in all patients. RESULTS: A 3q deletion was detected in one patient; two unrelated clones with +14 and +11 were present in another patient who had a cryptic 5q31 deletion as disclosed by FISH; both patients had a mild clinical course. The 5q31 deletion was shown to involve the eosinophilic lineage and not the lymphoid cells. No chromosome abnormalities were found by karyotyping or interphase FISH in the remaining 4 cases. In two of these cases the clinical course was aggressive, with progressive leukocytosis and marked splenomegaly in one patient, central nervous system and cardiac involvement as well as bone marrow failure in the other. INTERPRETATION AND CONCLUSIONS: The 3q deletion, +11 and +14, and a cryptic 5q31 deletion involving the cells of the eosinophilic lineage are three novel chromosome abnormalities occurring in HES. We did not find a correlation between evolving or aggressive disease and the presence of chromosome anomalies. Our data confirm that HES is a clinically and biologically heterogeneous condition and suggest that more cases need to be studied to identify clinically significant chromosome changes in this rare condition. Some patients may benefit from treatment with interferon. PMID- 10800165 TI - Diagnosis of essential thrombocythemia at platelet counts between 400 and 600x10(9)/L. Gruppo Italiano Malattie Mieloproliferative Croniche(GIMMC). AB - BACKGROUND AND OBJECTIVE: Diagnostic criteria for essential thrombocythemia (ET) remain essentially negative, that is, exclusion of other myeloproliferative diseases and causes of reactive thrombocytosis. A platelet count above 600x10(9)/L is still generally considered an absolute diagnostic criterion although new protocols for positive diagnostic criteria have recently been proposed, reducing the stringency of a definite platelet limit. This study demonstrates that a platelet count 600x10(9)/L is not a reliable diagnostic criterion for ET, especially in the early stages. DESIGN AND METHODS: An ongoing retrospective study by the GIMMC analyzed 2,316 ET patients diagnosed between 1986 and 1995. Of these 2,316 patients, diagnosed according to the PVSG criteria, 68 had a platelet count 600x10(9)/L and were analyzed separately; 37 out of 68 were excluded from this analysis because of a follow-up shorter than 2 years and/or because of treatment with myelosuppressive agents. The remaining 31 patients were the subjects of our study. RESULTS: After a median follow-up of 4.56 years (range 2-9.6 years) none of the 31 patients had a spontaneous decrease of platelets to the normal range. Transformation to a different chronic myeloproliferative disorders was never observed and no patient developed a condition known to produce reactive thrombocytosis. During follow-up, 23 patients (74%) were treated with anti-aggregating drugs, mainly aspirin. The disease did not evolve into acute leukemia in any patient, 1 had a thrombotic event and none presented hemorrhagic episodes. Median platelet count during follow-up was 534x10(9)/L (range 398-997x10(9)/L). INTERPRETATION AND CONCLUSIONS: Long term follow-up has documented that our 31 patients were correctly diagnosed as having ET, although platelet count was 600x10(9)/L. Our patients were probably in a early phase of their disease and following updated PVSG criteria would have been misdiagnosed leading to incomplete recognition of the natural history of the disease. Further, because an early diagnosis could also have a clinical relevance, our results outline the need for new criteria for the diagnosis of ET. The exclusion of patients with a platelet count between 400 and 600x10(9)/L may prevent patients, nevertheless at risk of vascular complications, from being treated. PMID- 10800166 TI - Coexistence of two distinct cell populations (CD56(+)TcRgammadelta(+) and CD56(+)TcRgammadelta(-)) in a case of aggressive CD56(+) lymphoma/leukemia. AB - BACKGROUND AND OBJECTIVE: Large granular lymphocytes derive from two major lineages: one expressing the CD3 surface antigen (T-lymphocytes), and the other lacking this marker (NK-cells). Although developmental overlaps between natural killer cells and T-cells have been described, malignancies derived from these two cell types are considered as distinct lymphoid disorders. DESIGN AND METHODS: We report the case of a 30-year old man affected by a lymphoma/leukemia syndrome presenting with hepatosplenic lymphoma which rapidly transformed into aggressive NK-leukemia. Extensive flow cytometry studies and molecular analysis were repeated during the course of the disease, and showed an unexpected changing pattern. RESULTS: At diagnosis, flow cytometry analysis showed the co-existence of two cell populations, one CD56(+), CD3(+), TcRgd(+), and the other CD56(+), CD3(-) and TcRgd(-). Molecular analysis showed that the TcR genes had the same clonally rearranged pattern involving b, g and d genes in both populations. At disease relapse and during the terminal refractory phase, only CD3(-) cells were present. INTERPRETATION AND CONCLUSIONS: This is an unusual case of CD56(+) aggressive lymphoma/leukemia characterized by the clonal expansion of two phenotypically different cell populations, variably balanced during the course of the disease. The presence of the same TcR genomic rearrangement suggests the origin from a common progenitor able to differentiate along both T- and NK pathways. PMID- 10800167 TI - Phase II study of 3-hour infusion of high dose paclitaxel in refractory and relapsed aggressive non-Hodgkin's lymphomas. Groupe d'Etude des Lymphomes de l'Adulte. AB - BACKGROUND AND OBJECTIVE: The first clinical studies of paclitaxel as a single agent for the treatment of relapsed or refractory low or intermediate grade non Hodgkin's lymphomas (NHL) yielded controversial results regarding the response rates observed, mainly related to the dose and schedule of administration used. To obtain additional data concerning the efficacy and toxicity of paclitaxel in intermediate and high grade NHL we initiated a phase II study using a 3-hour infusion of high doses of paclitaxel. DESIGN AND METHODS: The eligibility criteria included patients with relapsed or refractory aggressive NHL, a performance status < or = 2 (WHO index), a platelet count > or = 100,000/microL, a neutrophil count > or = 2,000/microL, measurable disease, and adequate hepatic function. Patients were excluded if they were infected with HIV, had a left ventricular ejection fraction < 50%, or prior peripheral neuropathy. Paclitaxel was administered as a 3-hour infusion at a dose of 250 mg/m2 every 3 weeks for a maximum of 6 courses. RESULTS: Of 45 eligible patients, 42 received a total 73 courses of paclitaxel. Forty patients were assessable for response (89%), and 42 for toxicity (93%). Six patients (15%) achieved a partial (n = 4) or a complete remission (n = 2). Responses were observed in intermediate grade (n = 4) as well as in high grade lymphoma (n = 2). The main factor influencing the response to paclitaxel was the median duration of response to previous chemotherapy regimens which was 3 times longer in patients who responded to paclitaxel (16.3 months) than in patients who did not respond to paclitaxel (5.2 months) (p<0.05). The most common serious side effects were related to the hematologic toxicity of paclitaxel, and included grade IV granulocytopenia in 20 cases (48%), grade III/IV thrombocytopenia in 14 cases (33%) and grade III-IV anemia in 13 cases (31%). INTERPRETATION AND CONCLUSIONS: Despite frequent manageable hematologic toxicity, paclitaxel is usually well tolerated at a dose of 250 mg/m2 given by a 3-hour infusion. However, the clinical efficiency as a single therapy seems modest in relapsed or refractory aggressive lymphoma. PMID- 10800168 TI - Dose-intensive melphalan with stem cell support (CM regimen) is effective and well tolerated in elderly myeloma patients. AB - BACKGROUND AND OBJECTIVE: Multiple myeloma (MM) typically afflicts elderly patients. High-dose therapy has recently been shown to lead to a better outcome than standard treatment, mainly in younger patients. The extent to which older subjects can benefit from intensified approaches without excessive toxicity is examined in this study. DESIGN AND METHODS: Between December 1994 and May 1997, 12 Italian Multiple Myeloma Study Group institutions entered 68 patients at diagnosis (median age 65) into an intensified chemotherapy regimen: cyclophosphamide (CY) 3 g/m(2) plus melphalan 60 mg/m(2) followed by peripheral blood progenitor cells (PBPC) and G-CSF (CM regimen). CY (day 0) and G-CSF were used to mobilize PBPC harvested by a single leukapheresis on day 10. Melphalan was infused on day 11. PBPC were kept unprocessed at 4 degrees C for 48 hours and reinfused on day 12. Three CM regimens were delivered at 6-month intervals. RESULTS: Sufficient PBPC to support the first CM cycle were available (median CD34(+) harvest: 4.9x10(6)/kg), but dropped significantly after the second (median CD34(+) harvest: 2x10(6)/kg) and the third (median CD34(+) harvest: 0.9x10(6)/kg). The median durations of severe neutropenia (absolute neutrophil count < 500 microL) were 3, 4, and 3 days, and those of severe thrombocytopenia (platelets < 25,000/microL) were 2.5, 2, and 1 days, after the first, second and third courses, respectively. The frequency of extramedullary toxicities was low. Treatment-related mortality (TRM) was 3% after the first CM, only. Complete remission (CR) was 14% after the first, 16% after the second and 27% after the third CM. After a median follow-up of 34 months (range 19-49 months), median event-free survival was 35.6 months. INTERPRETATION AND CONCLUSIONS: These results indicate that dose-intensity of melphalan can be increased by reinfusing PBPC with acceptable low toxicity. The combination of CY and melphalan followed by PBPC is an effective chemotherapy for elderly myeloma patients. Repeated melphalan infusion hampered subsequent CD34(+) harvests. PMID- 10800169 TI - Differential aspects of the glycoprotein Ib-von Willebrand factor axis in human and pig species. AB - BACKGROUND AND OBJECTIVE: The role of glycoprotein Ib (GPIb) in platelet adhesion to subendothelium is well established in human species. However, the interaction of GPIb and von Willebrand factor (VWF) in a widely used experimental model in thrombosis research, that of the pig, has not been clearly elucidated. We investigated the differences between human and pig species in the GPIb/VWF axis in several ways. DESIGN AND METHODS: Standard aggregometry and perfusion studies with circulating blood were applied to isolated platelets or to blood reconstituted with isolated platelets, VWF and red blood cells from the different species. Platelet aggregation to VWF in the presence of either ristocetin or botrocetin was tested. RESULTS: Human VWF and ristocetin did not agglutinate pig platelets. However, botrocetin was capable of agglutinating pig platelets. In perfusion studies (800 s(-1), 10 min), washed platelets from both species were suspended in albumin solutions containing human VWF (hVWF) or porcine VWF (pVWF) and red blood cells from the corresponding species. Reconstituted blood with high concentrations of pVWF (> or =0.25 U/mL) caused severe thrombocytopenia during the perfusion procedure when added to human platelets. Nevertheless, lower concentrations (< or =0.1 U/mL) promoted the formation of large aggregates. Under our experimental conditions, hVWF poorly supported pig platelet adhesion. INTERPRETATION AND CONCLUSIONS: In conclusion, pVWF may support human platelet adhesion and even promote aggregation, while hVWF can only partially facilitate pig platelet adhesion. Minimal concentrations of pVWF could facilitate the interaction of human platelets with subendothelium, increasing their adhesive and aggregating capabilities. Understanding the molecular differences of the GPIb-VWF axis in different species may prove useful for developing therapeutic strategies aimed at preventing excessive platelet deposition on damaged vascular surfaces. PMID- 10800171 TI - Experience of a single Italian center in genetic counseling for hemophilia: from linkage analysis to molecular diagnosis. AB - BACKGROUND AND OBJECTIVE: We describe our three year experience in genetic counseling at the Castelfranco Veneto Hemophilia Center, Italy. DESIGN AND METHODS: A total of 258 individuals were involved in the study of 142 females. These formed 40 families with hemophilia A and 6 families with hemophilia B. Following pedigree analysis, the FVIII inversion was first examined in severe hemophilia A patients by polymerase chain reaction (PCR) analysis. DNA polymorphisms were used to track the affected gene through the remaining families. In uninformative cases, we initiated analysis of the FVIII or FIX gene coding region by conformation sensitive gel electrophoresis and DNA sequencing to identify the mutation responsible for the disease. RESULTS: The FVIII gene inversion was present in 16 of the 32 patients (50%) affected by severe hemophilia A and was informative for 44 females. For hemophilia A, 45 cases (55%) were informative by linkage analysis, however 37 (45%) were uninformative because of lack of key individuals, homozygosity, or sporadic disease. Information from extragenic linked polymorphisms alone was present in 9 cases (6%). For hemophilia B, linkage analysis was informative in only 50% of females (8 out of 16). To date, nine mutations have been identified in patients with hemophilia A and three in patients with hemophilia B. Six novel missense mutations in hemophilia A are discussed briefly. INTERPRETATION AND CONCLUSIONS: Using this approach we are now able to offer accurate genetic analysis to virtually all families with hemophilia. PMID- 10800170 TI - Quantification of D-dimer using a new fully automated assay: its application for the diagnosis of deep vein thrombosis. AB - BACKGROUND AND OBJECTIVE: A D-dimer assay can be helpful to rule out thromboembolism provided it is sensitive, reliable, fast and easy to perform. Tests based on the ELISA methodology have a high diagnostic sensitivity, and are therefore adequate for excluding deep venous thrombosis (DVT). The drawbacks are their long assay times, unsuitability to be run on single samples and cost. New methods have been developed, based either on the same principle, by immunofiltration or by microlatex immunoturbidimetric assays which seem to reach the high sensitivity and negative predictive value (NPV) required, but allowing fast and quantitative single sample analysis. The aim of this work was to evaluate one rapid test, a fully automated quantitative assay (IL Test D-dimer, run on an ACL 7000 coagulation analyzer, Instrumentation Laboratory). DESIGN AND METHODS: We compared the diagnostic value of IL Test for DVT with that of an ELISA (Dimertest Gold EIA Agen Biomedical Limited, Acacia Ridge, Australia). Eighty-six patients (43 men, 43 women, mean age: 61 years) showing DVT symptoms formed the population for this non-randomized controlled trial in a referral center. The diagnosis of DVT based on the clinical history, was confirmed by serial compression ultrasonography (CUS) with Doppler flow in 62 patients. RESULTS: The IL Test D-dimer proved to be rapid, automated and well suited for individual tests with a good reproducibility in three control plasmas with different concentrations of D-dimer (coefficient of variation range 0.54-3. 87%). Its performance was comparable to that of the Dimertest Gold EIA, as indicated by the areas under the receiver operating characteristic curves (Dimertest Gold EIA 0.748; IL Test D-dimer 0.70). On the basis of kappa coefficients, there was a good concordance between the Dimertest Gold EIA and IL Test D-dimer when the receiver operating characteristic (ROC) curves suggested cut-offs were used. The sensitivity (98.3%) and NPV (88.9%) shown by IL Test D-dimer are comparable or even better than those obtained for EIA (95%, 80%, respectively). INTERPRETATION AND CONCLUSIONS: This study shows that the new method can be included in prospective clinical trials to test the utility of D-dimer measurement in combination with other non-invasive diagnostic procedures in the management of DVT diagnosis. PMID- 10800172 TI - Unrelated donor bone marrow transplantation as treatment for chronic myeloid leukemia: the Spanish experience. The Chronic Myeloid Leukemia Subcommittee of the GETH. Grupo Espanol de Trasplante Hemopoyetico. AB - BACKGROUND AND OBJECTIVE: To analyze the results of unrelated bone marrow transplantation (UDBMT) as treatment for chronic myeloid leukemia (CML) in Spain. DESIGN AND METHODS: Eighty-seven consecutive UDBMT performed in 9 centers between October 1989 and February 1998 were evaluated. This represents more than 95% of UDBMT for CML performed in adult transplant centers in Spain during this period. The patients' median age was 31.5 years (range, 12-49). The median interval from CML diagnosis to UDBMT was 30 months (range, 3-160). Seventy-nine percent of transplants were performed during the first chronic phase (1CP). RESULTS: Actuarial probability of survival and disease-free survival at 4 years for the whole series was 24% (95% confidence interval [CI]: 14%-34%) and 20% (CI: 10% 30%), respectively. The cumulative incidence of relapse and transplant-related mortality (TRM) was 7% (CI: 4%-10%) and 71% (CI: 60%-82%), respectively. The main causes of death were graft failure (n=7), infection (n=23), and graft-versus-host disease (GvHD) (n=25). The actuarial probability of acute GvHD grade II-IV and grade III-IV was 56% (CI:46%-66%) and 36% (CI: 26%-36%), respectively. The cumulative incidence of extensive chronic GvHD was 18% (CI: 9%-27%). Univariate analyses showed that the pre-transplant factor with the highest influence on survival was disease status at transplant (30% in 1CP vs. 0% in advanced phases; p=0.0001). Other pre-transplant factors influencing survival among patients in 1CP were: patient's age (older than 30 years 11% vs. 48%), interval diagnosis transplantation (longer than 2 years 17% vs. 55%), donor type (HLA, B, DRB1 identical 32% vs. 25%), CMV serologic status (donor and recipient negative 63% vs. 24%), year of transplantation (before 1995 19% vs. 40%), and conditioning regimen (cyclophosphamide plus total body radiation 40% vs. 16%). The main risk factors had a cumulative effect on survival. Thus, probability of survival ranged from 66% (CI: 39%-93%) in patients in 1CP, under 40 years of age, transplanted from an HLA, A, B, DRB1 identical donor during the first two years after diagnosis, to 0% in those with three or more risk factors. INTERPRETATION AND CONCLUSIONS: This experience shows that UDBMT used to have a high TRM that has progressively decreased along the years. At the present time, the results are encouraging, particularly when UDBMT is performed under favorable conditions. PMID- 10800173 TI - Systemic inflammatory pseudotumor, an unusual cause of fever of unknown origin mimicking a malignant lymphomatous process: case-report and review of the literature. AB - BACKGROUND AND OBJECTIVE: In recent years, a new pathologic condition Eth called inflammatory pseudotumor Eth has been occasionally described to involve many different tissues or organs. However, evidence for systemic involvement is lacking and a review of the topic potentially useful, also considering the clinical features mimicking hematologic malignancies or other clinically relevant conditions. We report a case of systemic inflammatory pseudotumor and review the literature concerning the etiopathogenesis, differential diagnosis and treatment modalities of this particular pathology. DATA SOURCES AND METHODS: The data source for this topic was MEDLINE , searching all fields for inflammatory pseudotumor, inflammatory pseudotumor and lymph node or fever of unknown origin, pertaining to humans, published between 1970 and 1999. RESULTS AND CONCLUSIONS: Inflammatory pseudotumor should be considered in the differential diagnosis of malignant hematologic processes and of fever of unknown origin. PMID- 10800174 TI - Lymphoid blastic crisis in Philadelphia chromosome-positive chronic granulocytic leukemia following high-grade non-Hodgkin's lymphoma A case report and review of literature. AB - In this paper we describe a case of a 65-year old man with a lymphoid blastic crisis of a chronic granulocytic leukemia occurring seven years after a palatine tonsillar non-Hodgkin's lymphoma treated with chemotherapy and radiation therapy. Bone marrow cytogenetic study demonstrated the presence of the typical t(9;22)(q34;q11) and the molecular biology study showed the p210 rearrangement (b2a2). The patient died within a few months, unresponsive to any treatment. This is the first case, described in literature, of a secondary chronic granulocytic leukemia onset with a lymphoid blastic crisis. The authors report the case and a literature review. PMID- 10800175 TI - Unusual presentation of hairy cell leukemia. PMID- 10800176 TI - Unexpected pulmonary embolism diagnosed by conventional computed tomography. PMID- 10800177 TI - Influence of sample storage time and temperature on lymphocyte subset counts using a FACScount system. PMID- 10800178 TI - Acute hepatomegaly with severe liver toxicity due to all-trans-retinoic acid. PMID- 10800180 TI - Myelofibrosis in myeloid malignancies with 3q26 cytogenetic abnormalities. PMID- 10800179 TI - Long-term disease-free acute myeloblastic leukemia with inv(16) is associated with PCR undetectable CBFbeta/MYH11 transcript. PMID- 10800181 TI - CD79b expression in B-cell chronic lymphocytic leukemia. PMID- 10800182 TI - Non-gastrointestinal malt lymphomas: a study of 10 cases and comparison with 27 patients with gastrointestinal MALT lymphoma. PMID- 10800183 TI - Spontaneous rupture of spleen during periferal blood stem cell mobilization in a patient with breast cancer. PMID- 10800184 TI - Human toxocarosis. Its seroprevalence in the city of La Plata. AB - Toxocara canis is very common in dogs throughout the world. It is the primary cause of visceral larva migrans (VLM) in humans. Soil contaminated with T. canis embryonated eggs is the main source of infection of man. Our objective was to describe Toxocara seroprevalence in humans in the city of La Plata associated with some determinants such asage, presence or absence of clinical manifestations and risk factors. Blood samples were collected at random from 156 patients of different sex and age, with and without clinical symptoms compatible with the disease. The diagnostic technique ELISA test was performed with the Bordier Affinity Products Commercial Kit, using excretory-secretory Toxocara antigen with a sensitivity higher than 90%. The values were positive in 39% of the studied population. In the analysis according to age, the younger group presented significant differences with respect to the older one (Chi-square p<0.05). Positive patients presented clinical symptoms compatible with the disease (84%), and 41% presented some risk factor. The level of positivity obtained indicates a certain risk of being infects mainly in patients younger than 15 years old. The authors agree that an early identification and treatment of VLM may save a life. PMID- 10800185 TI - Microhabitat preferences of Biomphalaria pfeifferi and Lymnaea natalensis in a natural and a man-made habitat in southeastern Tanzania. AB - Schistosoma mansoni is an important human parasitic disease which is widespread throughout Africa. As Biomphalaria pfeifferi snails act as intermediate host, knowledge of their population ecology is an essential prerequisite towards understanding disease transmission. We conducted a field study and assessed the density and microhabitat preferences of B.pfeifferi in a natural habitat which was a residual pool of a river. Repeated removal collecting revealed a density of 26.6 [95% confidence interval (CI): 24.9-28.3] snails/m2. B. pfeifferi showed microhabitat preferences for shallow water (depths: 0-4cm). They were found most abundantly close to the shoreline (distances: 0-40cm), and preferred either plant detritus or bedrock as substratum. Lymnaea natalensis, a snail which may act as a host for human Fasciola gigantica, also occurred in this habitat with a density of 34.0 (95% CI: 24.7-43.3) snails/m2, and preferred significantly different microhabitats when compared to B.pfeifferi. Microhabitat selection by these snail species was also investigated in a man-made habitat nearby, which consisted of a flat layer of concrete fixed on the riverbed, covered by algae. Here, B.pfeifferi showed no preference for locations close to the shoreline, probably because the habitat had a uniform depth. We conclude that repeated removal collecting in shallow habitats provides reliable estimates of snail densities and that habitat changes through constructions may create favourable microhabitats and contribute to additional disease transmission. PMID- 10800186 TI - Variation in cuticular hydrocarbons among strains of the Anopheles gambiae sensu stricto by analysis of cuticular hydrocarbons using gas liquid chromatography of larvae. AB - Cuticular hydrocarbons of larvae of individual strains of the Anopheles gambiae sensu stricto were investigated using gas liquid chromatography. Biomedical discriminant analysis involving multivariate statistics suggests that there was clear hydrocarbon difference between the Gambian(G3), the Nigerian (16CSS and, its malathion resistant substrain, REFMA) and the Tanzanian (KWA) strains. The high degree of segregation (95%) in hydrocarbons among the four strains investigated indicates that further analysis is needed to enable understanding of hydrocarbon variation in samples of An. gambiae especially from areas where these populations co-exist. PMID- 10800187 TI - Oswaldotrema nacinovici gen. nov. sp. nov. (Digenea: Philophthalmidae) from Numenius phaeopus (Aves: Scolopacidae) in Brazil. AB - A new genus, Oswaldotrema gen. nov. is proposed. Oswaldotrema nacinovici sp. nov. is descibed from Numenius phaeopus Latham, 1790. Differentiation from the other related genera, namely Philophthalmus, Pygorchis, Proctobium, Parorchis, Echinostephila, Cloacitrema, Pittacium, Ophthalmotrema, Skrjabinovermis and Paratrema, was based on morphological characters, mainly on those referring to the body surface, body shape, head, esophagus, pharynx, acetabulum, vitellaria, vitelline reservoir and seminal vesicle. PMID- 10800188 TI - Studies on populations of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in Brazil. AB - Studies were performed on five Brazilian populations of Lutzomyia longipalpis: Salvaterra (PA), Sao Jose do Ribamar (MA), Caninde (CE), Natal (RN) and Gruta da Lapinha, Lagoa Santa (MG). No morphological differences were observed that could distinguish between these populations. Homogeneity tests showed that the allopatric populations display a certain heterogeneity and that the sympatric populations, with different patterns of spots, are homogeneous. The Student Newman-Keuls test, represented by Euler-Venn diagrams, showed a disjunction between the populations from the north/northeast and the one from Gruta da Lapinha. Genetic distances between the four populations (excluding the Caninde population) were within the range of intrapopulational differences. The Gruta da Lapinha population displayed a heterozygotic deficiency that could be a consequence of high levels of inbreeding due to cryptic habits of living in a small cave. These results do not favor the hypothesis of a L. longipalpis species complex in Brazil, and the species should be considered high polymorphic. PMID- 10800189 TI - Direct methods for detecting picorna-like virus from dead and alive triatomine insects. AB - In this work we report four different destructive and non-destructive methods for detecting picorna-like virus particles in triatomines. The methods are based on direct observation under transmission electron microscope and they consist of four ways to prepare samples of presumable infected material. The samples are prepared processing dead or alive insect parts, or even dry or fresh insect feces. The methods can be used as analytical or preparative techniques, for quantifying virus infection and checking virus integrity as well. In this work the four methods are applied in order to detect Triatoma virus (TrV) particles in T. infestans colonies. PMID- 10800190 TI - Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography. AB - The invasion of the erythrocyte by Plasmodium falciparum depends on the ability of the merozoite to move through the membrane invagination. This ability is probably mediated by actin dependent motors. Using affinity columns with G-actin and F-actin we isolated actin binding proteins from the parasite. By immunoblotting and immunoprecipitation with specific antibodies we identified the presence of tropomyosin, myosin, a-actinin, and two different actins in the eluate corresponding to F-actin binding proteins. In addition to these, a 240-260 kDa doublet, different in size from the erythrocyte spectrin, reacted with an antibody against human spectrin. All the above mentioned proteins were metabolically radiolabeled when the parasite was cultured with 35S-methionine. The presence of these proteins in P. falciparum is indicative of a complex cytoskeleton and supports the proposed role for an actin-myosin motor during invasion. PMID- 10800191 TI - A comparison of three DNA extractive procedures with Leptospira for polymerase chain reaction analysis. AB - Three DNA extraction methods were evaluated in this study: proteinase K followed by phenol-chloroform; a plant proteinase (E6870) followed by phenol-chloroform; and boiling of leptospires in 0.1 mM Tris, pH 7.0 for 10 min at 100 degrees C, with no phenol treatment. Every strain treated with proteinase K or E6870 afforded positive polymerase chain reaction (PCR) reaction. On the other hand, from five strains extracted by the boiling method, three did not feature the 849 bp band characteristic in Leptospira. We also evaluated by RAPD-PCR, DNAs from serovars isolated with proteinase K and proteinase 6870 with primers B11/B12. Each of the DNA samples provided PCR profiles in agreement with previous data. Moreover, the results with E6870 showed less background non-specific amplification, suggesting that removal of nucleases was more efficient with E6870. The limit for detection by PCR using Lep13/Lep14 was determined to be 10(2) leptospira, using the silver stain procedure. PMID- 10800192 TI - Plasmodium yoelii: identification of a gene encoding a putative ADP-ribosylation factor-1 GTPase-activating protein, PyAG1. AB - The PyAG1 gene, identified by the screening of a Plasmodium yoelii genomic DNA library with a rhoptry-specific Mab, encodes a protein with a zinc finger structure immediately followed by the consensus sequence of the Arf GAP catalytic site. The serum of mice immunized with the recombinant protein recognized specifically the rhoptries of the late infected erythrocytic stages. Blast analysis using the Genbank database gave the highest scores with four proteins presenting an Arf1 GAP activity. If presenting also this activity, the PyAG1 protein could be involved in the regulation of the secreted protein vesicular transport and, consequently, in the rhoptry biogenesis. PMID- 10800193 TI - Significance of schistosomal granuloma modulation. AB - Hepatic Schistosoma mansoni periovular granulomas undergo changes in size, cellular composition and appearance with time. This phenomenon, known as "immunological modulation", has been thought to reflect host immunological status. However, as modulation has not been observed outside the liver, participation of local factors, hitherto little considered, seems crucial. Components of the extracellular matrix of periovular granulomas of the mouse were particularly studied in three different organs (liver, lung and intestine) and during three periods of infection time (acute, intermediate and chronic) by means of histological, biochemical and immunofluorescence techniques, while quantitative data were evaluated by computerized morphometry, in order to investigate participation of local factors in granuloma modulation. Results confirmed modulation as a exclusively hepatic phenomenon, since pulmonary and intestinal granulomas, formed around mature eggs, did not change size and appearance with time. The matricial components which were investigated (Type I, III and IV collagens, fibronectin, laminin, proteoglycans and elastin) were found in all granulomas and in all organs examined. However, their presence was much more prominent in the liver. Elastin was only found in hepatic granulomas of chronic infection. The large amount of extracellular matrix components found in hepatic granulomas was the main change responsible for the morphological aspects of modulation. Therefore, the peculiar environment of the liver ultimately determines the changes identified in schistosomal granuloma as "modulation". PMID- 10800194 TI - Aotus infulatus monkey is susceptible to Plasmodium falciparum infection and may constitute an alternative experimental model for malaria. AB - Aotus is one of the WHO-recommended primate models for studies in malaria, and several species can be infected with Plasmodium falciparum or P. vivax. Here we describe the successful infection of the species A. infulatus from eastern Amazon with blood stages of P. falciparum. Both intact and splenectomized animals were susceptible to infection; the intact ones were able to keep parasitemias at lower levels for several days, but developed complications such as severe anemia; splenectomized monkeys developed higher parasitemias but no major complications. We conclude that A. infulatus is susceptible to P. falciparum infection and may represent an alternative model for studies in malaria. PMID- 10800195 TI - Biological screening of Brazilian medicinal plants. AB - In this study, we screened sixty medicinal plant species from the Brazilian savanna ("cerrado") that could contain useful compounds for the control of tropical diseases. The plant selection was based on existing ethnobotanic information and interviews with local healers. Plant extracts were screened for: (a) molluscicidal activity against Biomphalaria glabrata, (b) toxicity to brine shrimp (Artemia salina L.), (c) antifungal activity in the bioautographic assay with Cladosporium sphaerospermum and (d) antibacterial activity in the agar diffusion assay against Staphylococcus aureus, Escherichia coli, Bacillus cereus and Pseudomonas aeruginosa. Forty-two species afforded extracts that showed some degree of activity in one or more of these bioassays. PMID- 10800196 TI - Study of Schistosoma mansoni isolates from patients with failure of treatment with oxamniquine. AB - After three successive treatments with oxamniquine the continuing elimination of Schistosoma mansoni eggs was observed in patients, who came from various regions of Brazil, with different clinical forms of schistosomiasis. The objective of the present study was to determine the experimental behaviour of five different S. mansoni isolates in Swiss Webster mice that were submitted to treatment with the same drug. The experimental group with failure of treatment showed higher mean number of surviving male worms when it was compared to the group without failure of treatment. These date suggest the possibility of resistance to oxamniquine. PMID- 10800197 TI - The central projection of cephalic mechanosensory axons in the haematophagous bug Triatoma infestans. AB - The projections of mechanosensory hairs located on the dorsal and lateral head of the adult haematophagous bug Triatoma infestans were analyzed by means of cobalt filling. Axons run into the anterior and posterior tegumentary nerve and project through the brain to the ventral nerve cord. The fibres are small in diameter and run as a fascicle. Some branches run into suboesophageal and prothoracic centres; others run as far as to the mesothoracic ganglion. These sensory projections resemble that of wind-sensitive head hairs of the locust. The functional role of this sensory system in this species is discussed. PMID- 10800198 TI - Immunological evaluation of human immunodeficiency virus infected individuals by flow cytometry. AB - Human immunodeficiency virus (HIV) infection heavily compromises the immune system. The decrease of the T cell CD4+ subset along the evolution to acquired immunodeficiency syndrome has been considered as a hallmark of HIV infection. In this paper we review some aspects of the immunopathology of HIV infection and discuss the importance of the flow cytometry for the evaluation of the T lymphocyte subsets in the follow-up of HIV infected children and adults, and for the monitoring of the immune reconstitution upon antiretroviral therapy. PMID- 10800199 TI - Detection of intracytoplasmic cytokines by flow cytometry. AB - Flow cytometry has been used as a powerful technique for studying cell surface antigen expression as well as intracellular molecules. Its capability of analyzing multiple parameters simultaneously on a single cell has allowed identification and studies of functional cell subsets within heterogeneous populations. In this respect, several techniques have been developed during the past few years to study cytokine-producing cells by flow cytometry in humans and several animal models. PMID- 10800200 TI - Applications of flow cytometry to hematopoietic stem cell transplantation. AB - Applications of flow cytometry to clinical and experimental hematopoietic stem cell transplantation (HSCT) are discussed in this review covering the following topics: diagnosis and classification of lymphohematologic disorders, quantitation of hematopoietic progenitors in the graft, lymphohematopoietic reconstitution following HSCT and animal models of human HSCT. At the end, the utilization of flow cytometry in clinical HSCT by Brazilian transplant centers is briefly reviewed. PMID- 10800201 TI - Procedures to characterize and study P2Z/P2X7 purinoceptor: flow cytometry as a promising practical, reliable tool. AB - The expression of P2Z/P2X7 purinoceptor in different cell types is well established. This receptor is a member of the ionotropic P2X receptor family, which is composed by seven cloned receptor subtypes (P2X1 - P2X7). Interestingly, the P2Z/P2X7 has a unique feature of being linked to a non-selective pore which allows the passage of molecules up to 900 Da depending on the cell type. Early studies of P2Z/P2X7 purinoceptor were exclusively based on classical pharmacological studies but the recent tools of molecular biology have enriched the analysis of the receptor expression. The majority of assays and techniques chosen so far to study the expression of P2Z/P2X7 receptor explore directly or indirectly the effects of the opening of P2Z/P2X7 linked pore. In this review we describe the main techniques used to study the expression and functionality of P2Z/P2X7 receptor. Additionally, the increasing need and importance of a multifunctional analysis of P2Z/P2X7 expression based on flow cytometry technology is discussed, as well as the adoption of a more complete analysis of P2Z/P2X7 expression involving different techniques. PMID- 10800202 TI - Flow cytometry in the study of cell death. AB - In this report we present a concise review concerning the use of flow cytometric methods to characterize and differentiate between two different mechanisms of cell death, apoptosis and necrosis. The applications of these techniques to clinical and basic research are also considered. The following cell features are useful to characterize the mode of cell death: (1) activation of an endonuclease in apoptotic cells results in extraction of the low molecular weight DNA following cell permeabilization, which, in turn, leads to their decreased stainability with DNA-specific fluorochromes. Measurements of DNA content make it possible to identify apoptotic cells and to recognize the cell cycle phase specificity of apoptotic process; (2) plasma membrane integrity, which is lost in necrotic but not in apoptotic cells; (3) the decrease in forward light scatter, paralleled either by no change or an increase in side scatter, represent early changes during apoptosis. The data presented indicate that flow cytometry can be applied to basic research of the molecular and biochemical mechanisms of apoptosis, as well as in the clinical situations, where the ability to monitor early signs of apoptosis in some systems may be predictive for the outcome of some treatment protocols. PMID- 10800203 TI - Expression and regulation of NMDA receptor subunit R1 and neuronal nitric oxide synthase in cortical neuronal cultures: correlation with cytochrome oxidase. AB - Our previous studies showed a differential distribution of the glutamatergic terminals in cytochrome oxidase-rich and -poor regions of the visual cortex. The NMDA type of glutamate receptors have been proposed to be involved in the activation of nitric oxide synthase to produce nitric oxide, the neurotransmitter. In the present study, we hypothesized that the expressions of glutamate receptor, NMDA receptors (NMDAR1) and neuronal nitric oxide synthase (nNOS) were colocalized and were also correlated with that of cytochrome oxidase (CO) in a subset of neurons. We used primary cultures of postnatal rat visual cortical neurons as a model system, so that we could examine both the somatic and dendritic expressions of these neurochemicals in individual neurons. We found a difference in the sequence of developmental expressions of NMDAR1, nNOS, CO, and Na(+)/K(+) ATPase. Triple labeling showed that all nNOS-positive neurons were immunoreactive for NMDAR1, and a subpopulation of them had high CO activity. The expression of NMDAR1 was positively correlated with CO activity. This is consistent with our previous finding that CO activity is strongly governed by excitatory glutamatergic synapses. After 40 hours of depolarizing potassium chloride treatment, CO activity was increased, and NMDAR1and nNOS levels were up regulated in parallel. One week of tetrodotoxin significantly decreased the expression of NMDAR1, nNOS, and CO activity. Our results demonstrate that NMDA receptors and nNOS do co-exist in a subset of neurons that have high CO activity and their expressions are under the control of neuronal activity. PMID- 10800204 TI - Corticosterone replacement restores normal morphological features to the hippocampal dendrites, axons and synapses of adrenalectomized rats. AB - A thorough evaluation of hippocampal dendrites, axons and synaptic contacts has not been undertaken following prolonged periods of absence of corticosteroids despite the marked granule cell loss which occurs in the dentate gyrus of adrenalectomized rats. Thus, we have applied morphometric techniques to analyse the dendrites of granule and pyramidal cells, the mossy fiber system, and the number and morphology of synapses between the mossy fibers and the excrescences of CA3 pyramidal cells in rats submitted to different periods of adrenalectomy. In addition, to search for the presence of neuritic reorganisation in the hippocampal formation once normal corticosteroid levels were re-established, we incorporated in this study a group of rats replaced with corticosterone one month after adrenalectomy. The results obtained in adrenalectomized rats showed a striking impoverishment of the dendrites of surviving granule cells, subtle alterations in the apical dendritic arborization of CA3 pyramidal cells and no changes in the apical dendrites of CA1 pyramidal cells. In addition, in adrenalectomized rats there was a progressive reduction in the total number of synapses established between mossy fibers and CA3 pyramids, as a consequence of a reduction in the volume of the suprapyramidal part of the mossy fiber system, and profound changes in the morphology of mossy fiber terminals and CA3 dendritic excrescences. A remarkable reorganisation of neurites was found to occur following the administration of low doses of corticosterone, completely reversing the adrenalectomy-induced synaptic loss and partially restoring the morphology of hippocampal axons and dendrites. These plastic mechanisms provide a sound structural basis for the reversibility of cognitive deficits observed after corticosterone administration to adrenalectomized rats. PMID- 10800205 TI - Glial cell responses, complement and apolipoprotein J expression following axon injury in the neonatal rat. AB - Immature motoneurons are highly susceptible to degeneration following axon injury. The response of perineuronal glia to axon injury may significantly influence neuronal survival and axon regeneration. We have examined the central reactions to neonatal facial nerve transection with emphasis on the expression of complement component C3 (C3) and the multifunctional apolipoprotein J (ApoJ). Axotomy was performed on one-day-old rats. Animals were perfused from eight hours to two weeks after the lesion. The astroglial marker, glial fibrillary acidic protein (GFAP) was increased from one day and the microglial marker OX-42 from two days after injury. ApoJ immunoreactivity was increased in axotomized neuronal perikarya and astroglial cells from one day postaxotomy, but no C3 immunoreactive profiles were found at any postoperative survival time. Cell proliferation as judged by bromodeoxyuridine labeling and immunoreactivity for the cyclin Ki-67 antigen (antibody MIB5) occurred only at two days after injury. Double immunostaining revealed that the vast majority of proliferating cells were microglia, although occasional cells double labeled astrocytes were found as well. Our results indicate that the non-neuronal response in neonatal animals differ from that of adult ones as follows: 1) microglia transform rapidly into phagocytes in parallel with the degeneration of axotomized neurons, 2) despite the presence of neuronal degeneration, no expression of C3 was found, and the upregulation of the expression of the complement C3 receptor (CR3) is delayed, 3) ApoJ is strongly upregulated in perineuronal astrocytes as well as in the axotomized motoneurons. The marked upregulation of ApoJ in both instances suggests a general role of this protein in the neuronal response to axotomy. PMID- 10800206 TI - Expression of nerve growth factor, p75, and the high affinity neurotrophin receptors in the adult rat trigeminal system: evidence for multiple trophic support systems. AB - We hypothesize that discrete trigeminal structures have the components required for autocrine regulation as well as redundant neurotrophin support systems. We examined the expression of nerve growth factor (NGF) and the low affinity (p75) and high affinity (trkA, trkB, and trkC) neurotrophin receptors in the trigeminal system of adult rats. Four sites were examined; the trigeminal ganglion, mesencephalic nucleus, principal sensory nucleus (PSN), and trigeminal motor nucleus. NGF was expressed by more than 60% of neurons in each area studied. NGF immunolabeling may have resulted from exogenous protein incorporated from the microenvironment or from NGF synthesized by the neuron per se. To resolve this issue, in situ hybridization for NGF mRNA was performed. The mRNA was expressed by 2/3 to 7/8 of neurons in trigeminal structures. Moreover, double-labeling studies showed that virtually every ganglion cell that was NGF-immunoreactive also expressed the NGF transcript. Neurotrophin receptors (p75 and trk isoforms) were expressed by more than 60% of the neurons in each trigeminal structure. The only exception was the PSN, where the receptors were expressed by fewer than half of the neurons. Taken together, these data imply that NGF must be elaborated by neurons that co-express both p75 and trkA. Therefore, each trigeminal structure has the machinery for autocrine/paracrine regulation, as well as the capacity for retrograde and/or anterograde trophic support. Furthermore, the co-expression of the specific trk isoforms indicates that trigeminal neurons are sensitive to more than one neurotrophin. PMID- 10800207 TI - Localization of retinoid binding proteins, retinoid receptors, and retinaldehyde dehydrogenase in the chick eye. AB - Retinoids have many functions in the eye, including, perhaps, the visual guidance of ocular growth. Therefore, we identified where retinoid receptors, binding proteins, and biosynthetic enzymes are located in the ocular tissues of the chick as a step toward discovering where retinoids are generated and where they act. Using antibodies to interphotoreceptor retinoid binding protein (IRBP), cellular retinol binding protein (CRBP), cellular retinoic acid binding protein (CRABP), cellular retinaldehyde binding protein (CRALBP), retinaldehyde dehydrogenase (RALDH), and retinoic acid receptors (RAR and RXR), we localized these proteins to cells in the retina, retinal pigmented epithelium, choroid and sclera of the chick eye. IRBP was detected in the photoreceptor layer and pigmented epithelium; CRBP was in the pigmented epithelium; CRABP was in amacrine and bipolar cells in the retina; CRALBP was in Muller cells, pigmented epithelium, choroid, and fibrous sclera; RALDH was in retinal amacrine cells, pigmented epithelium, and choroid; RAR was in amacrine cells, choroid, and chondrocytes and fibroblasts in the sclera; and RXR was in amacrine and ganglion cells, bipolar cell nuclei, choroid, and chondrocytes. We also found that the growth-modulating toxins colchicine and quisqualate destroyed selectively different subsets of CRABP containing amacrine cells. We conclude that the distribution of proteins involved in retinoid metabolism is consistent with a role of retinoids not only in phototransduction, but also in maintenance of cellular phenotype and visual guidance of ocular growth. PMID- 10800208 TI - Waiting for Australia to act. PMID- 10800209 TI - Health care in the Czech Republic. PMID- 10800210 TI - What are the special needs of chronically ill young people? AB - Although significant advances have been made in the treatment of serious disease, there remains much scope for assisting young people in adjusting to life with a chronic medical condition. Commonly, chronically ill young people experience lower emotional well being than their healthy peers. Conventional approaches to promoting emotional well being have involved referring young people and their families to an appropriate public mental health service or psychologist/psychiatrist in private practice. However, there is increasing interest in the use of peer support programs. Support groups such as the ChIPS program aim to promote positive adjustment to chronic illness by bring together young people facing similar circumstances. It is maintained that by increasing connections between chronically ill young people, emotional well being can be enhanced. PMID- 10800211 TI - How to avoid a looming epidemic. PMID- 10800212 TI - A new perspective to long-term weight management. Is there a better way? AB - BACKGROUND: To manage the increasing rate of excess weight and obesity, individual health care providers and many health promotion programs have focused on encouraging people to lose weight, and/or achieve a certain weight, for optimal health. The focus has been on the person's weight, in other words, on the 'end point'. OBJECTIVE: This article provides a theoretical background to the need to think of the issue of weight management in a new way. DISCUSSION: In an environment that has been described as 'obesogenic', past and current strategies have not succeeded in creating positive change. The time has come for a new 'process orientated', more holistic focus. PMID- 10800213 TI - Tips for long-term weight management. AB - BACKGROUND: Finding the best way to help our patients achieve and maintain the most healthy weight they can is not simple. There are many factors that can contribute to an individual exceeding his or her most healthy weight, including a complex mix of behavioural and biological variables. OBJECTIVE: This article explores some of the important strategies people need to employ to achieve and maintain the healthiest weight that is possible for them. DISCUSSION: Unfortunately, there is no standard advice for weight management. Rather, it is about helping each patient find a balance between the amount of energy consumed in food and drink and the amount of energy expended. We need to help our patients gather all the relevant information, and find the best way of assisting them to put that knowledge together. The best plan of action also involves helping our patients work on why they are having difficulty achieving and maintaining a healthy weight, having the right perspective, and helping them develop some new skills to achieve long term change. PMID- 10800214 TI - Does 'getting physical' help weight control? AB - BACKGROUND: It may be time to embrace new attitudes in relation to health, weight loss and physical activity. While Australians spend around $1 million every day in attempts to lose weight, the number of people who are over their most healthy weight is increasing. OBJECTIVE: This article discusses practical ways to encourage patients to be more active by emphasising optimal health for each individual. DISCUSSION: Focusing on weight loss as the primary motivation to exercise, from a health and aesthetic perspective, may be the problem. Understanding the barriers to people exercising and focusing on the health benefits rather than the weight loss component is a useful way to get more people to exercise. PMID- 10800215 TI - Obesity in childhood and adolescence. AB - BACKGROUND: Obesity in childhood and adolescence is increasing in developed countries around the world. Management is often difficult and can be unsuccessful. OBJECTIVE: The epidemiology, causes and complications of obesity in childhood and adolescence are described, and a general clinical approach to weight management is provided. DISCUSSION: In general, a comprehensive history and examination can elicit medical causes of obesity. This paper does not seek to expand on medical causes but rather focuses on the clinical issues dealing with management of the overweight child or adolescent. This approach relies on actively engaging the child or young person, as well as involving the entire family in the process. Weight management consists of a combination of healthy eating behaviour and engaging in physical activity. PMID- 10800217 TI - Middle aged spread. Is it inevitable? PMID- 10800216 TI - The recognition and management of eating disorders. AB - BACKGROUND: Eating disorders--anorexia nervosa and bulimia nervosa--are increasingly recognised in our community, and appear to be increasing in frequency and affecting girls at a younger age, though the disorder is not restricted to females. OBJECTIVE: This article provides a current overview of eating disorders and their management. DISCUSSION: There continues to be controversy over the reason for the increase in eating disorders. Changes in societal ideas and pressures, as well as the massive increase this century in the role of the mass media, have been highlighted as having a likely role. Early engagement and a multifaceted approach are recommended. PMID- 10800218 TI - Anxiety disorders. Recognition and management. AB - BACKGROUND: Anxiety disorders are common and disabling. Yet they often go unnoticed: sufferers do not always complain of symptoms, and when they do they often present with primarily somatic concerns. Also, they are sometimes dismissed as being 'just anxious', 'very shy', or 'worriers' rather than being recognized as suffering from very treatable disorders. OBJECTIVE: To briefly describe the assessment, diagnosis, differential diagnosis, and management of the major anxiety disorders. DISCUSSION: Anxiety disorders are common and treatable. Cognitive behavioural therapy and medications are the two main treatment modalities. Ways of increasing skills with psychological techniques are also discussed. PMID- 10800220 TI - Clinical breast examination PMID- 10800219 TI - Clinical breast examination. A contentious issue in screening for breast cancer. AB - BACKGROUND: Breast cancer is the commonest cancerous cause of mortality in Australian women. It is now well established that mammography is effective in reducing this mortality through screening. To this end all states of Australia have established breast cancer screening programs using mammography as their investigative modality. The purpose of this review is to determine the value of clinical breast examination as another screening tool available to general practitioners. OBJECTIVE: We identified and examined a large amount of literature relating to this subject through Medline search, the Cochrane Library and hand searching (using keywords 'breast neoplasm', 'screening', 'breast self examination' and 'clinical breast examination'). The review of articles was undertaken by both authors, one of whom has extensive experience as a breast physician. DISCUSSION: Clinical breast examination for symptomatic women shows a true positive rate or sensitivity of 85%, a false positive rate of 20% and specificity of 80% for detecting the probability of cancer. The positive predictive value of this test is 81% and the negative predictive value is 84%. The sensitivity would be lower for asymptomatic women. CONCLUSION: Much has been written about screening for breast cancer. However, the absence of comprehensive studies leaves the evidence for CBE somewhat in dispute. In the absence of any harm from the procedure, it is difficult to support the currently accepted Australian evidence based guidelines which discourage the practice of CBE. It would seem more prudent given the disagreement in the literature and the acceptability of CBE to both GPs and patients to consider that there is value in using CBE as a screening tool. PMID- 10800221 TI - A tender swollen eyelid. PMID- 10800222 TI - A puzzling rash. Is a skin biopsy a useful diagnostic tool? AB - The diagnosis of a puzzling skin rash may require the clinician to provide a well taken biopsy specimen with an accompanying detailed description of the rash. The pathologist needs to be cognizant of the clinical manifestation of skin eruptions and to correlate the histopathological findings with the clinical features. PMID- 10800223 TI - A persistent wheeze. PMID- 10800224 TI - Lateral forefoot pain. PMID- 10800225 TI - Stress and cancer. PMID- 10800226 TI - Dealing with child protection problems. PMID- 10800227 TI - It takes two to tango. PMID- 10800228 TI - Where's the common ground? A survey of hospitals, community health centres and divisions of general practice in Australia. AB - BACKGROUND: Divisions of general practice are key structures for integration between general practice and other health services in Australia. AIM: To compare the views of divisions of general practice toward integration of care with those of hospitals and community health services. METHOD: Representative national samples of public hospitals and community health centres (CHCs) and a census of divisions of general practice (DGP) were surveyed on their current collaborations and links as well as barriers to and factors that enhance integration between general practitioners and other health services. RESULTS: There is wide agreement on the need for greater integration. Personal links (via letter, phone and face to face) were thought to be useful. However, general practice liaison officers were seen as especially useful. All organizations rated different accountabilities and responsibilities as a highly significant barrier. Resources, structures for collaboration and high level organisational support were rated as being more useful in enabling greater integration. CONCLUSION: Formalizing collaboration will require changes to funding and accountability. However there is also a need for cultural change to support greater integration of patient care between general practice and both hospitals and community health services. PMID- 10800229 TI - Who cares? A focus group study on follow up care after discharge from hospital. AB - AIM: To ascertain general practitioners', patients' and hospital staff's perspectives on post discharge problems and other problems related to hospital liaison in the Darwin urban area. To identify stakeholders who have a role in addressing these problems and develop locally appropriate responses to improve post hospital discharge patient care. METHOD: A qualitative study using three focus groups was conducted with seven GPs, six ex-patients of the study hospital and 11 hospital staff. The problems and proposed strategies identified by these groups were then fed back to stakeholders for comment in order to generate recommendations to improve post discharge care. RESULTS: The main themes generated were: lack of recognition of the potential role of the GP and patient by the hospital; poor communication between the hospital, GPs and patients; and, lack of resources to improve information flow. Specifically, 'not being listened to' by hospital doctors, nurses and GPs was a major theme of the patient focus group and was equated with a 'not caring' attitude by some hospital staff and GPs. Twelve strategies were generated by the focus groups with considerable overlap of issues. The main strategies were aimed at GP liaison by providing GPs and patients with timely, adequate and comprehensible information as part of their 'duty of care'. General practitioners, however, need to provide legible, relevant and concise referral letters to facilitate this process. CONCLUSIONS: This study highlights the need for improved communication between the GP and the hospital to redress the lack of recognition of GPs. The appointment of a GP/hospital liaison officer in the study hospital could address this issue as well as others of patient empowerment and advocacy, and partnerships in health care. This project has also highlighted the importance of consulting patients and the value of their perspective to complement GP and hospital viewpoints. PMID- 10800230 TI - Care for older persons. Are divisional programs meeting needs? AB - BACKGROUND: A national forum on healthy ageing was convened for divisions of general practice in November 1998. OBJECTIVE: Views of delegates were sought regarding divisional programs for older people related to the following areas: working in collaboration; integration; the context of the older person; ageism; and structural barriers. These areas were felt to influence success of current prospects for future programs for older people. DISCUSSION: Further communication, collaboration and integration appears essential to realise the potential that general practice offers to programs for older people. Issues from the forum are discussed in relation to relevant National General Practice Strategy Review and the changes introduced into the Medical Benefits Schedule in November 1999. Consideration of these themes will aid future divisional programs for older people. PMID- 10800231 TI - Antibiotic prophylaxis in dentistry. PMID- 10800232 TI - Tackling tooth erosion. PMID- 10800233 TI - Transfer of radiographs questioned. PMID- 10800234 TI - Persistent impairment of taste associated with terbinafine. AB - A second case of persistent taste disturbance associated with terbinafine is described. Taste disturbance associated with this drug is reviewed and a table is provided listing the more common drugs associated with taste disturbance. PMID- 10800235 TI - The first orthodontic diploma. AB - The advent of the Inter-Collegiate Membership in Orthodontics in the year 2000 will mark the end of an orthodontic diploma specific to the Royal College of Physicians and Surgeons of Glasgow. The Diploma in Dental Orthopaedics (RCPS) was the first orthodontic diploma in the United Kingdom. The fact that such a diploma has existed first as a DDO and latterly as the MDO (Membership) for some 50 years is remarkable in view of the relative infancy of most dental specialties and the fact that the General Dental Council has instigated a specialist register in orthodontics as recently as 1998. It was 1965, before other specialty diplomas became available in the British Isles, with the introduction of the FFD by the Royal College of Surgeons in Ireland. PMID- 10800236 TI - Effects of evidence-based treatment and consent on professional autonomy. AB - Recent high profile cases like Wisheart (Bristol babies) has placed healthcare professional autonomy under both ethical and legal scrutiny, reflected by the growing number of civil cases brought by the public. The aim of this paper is to investigate the effect of evidence-based treatment on the legal requirements of consent, when the treatment transcends the therapeutic boundary. The potential effect on the autonomy of healthcare professionals is also discussed. The study revealed that although consent remains the cornerstone of defence in many areas of therapeutic treatment, it is open to significant legal challenge in the grey area between therapeutic evidence-based treatment and non-evidenced-based procedures. Consent remains the cornerstone of defence for therapeutic procedures. However, it is less certain with innovative treatment or procedures that transcend the therapeutic boundaries and are shown to be non-evidence-based. PMID- 10800237 TI - The British Nutrition Foundation Oral Task Force report--issues relevant to dental health professionals. AB - A recent report on diet and oral health by the British Nutrition Foundation reviews the anatomy, microbiology and pathology of a number of oral diseases including dental caries, tooth-wear, oral cancer, periodontal disease and enamel defects. The role of nutritional factors in the aetiology and prevention of these oral diseases is discussed. The report states that improvements in the levels of caries in the UK are halting and remain unacceptably high in some 'at risk groups'--including the socially deprived. It states that a two-pronged attack, i.e. reduced frequency of consumption of sugary foods and use of fluoride is necessary to address the problem. To address the increased prevalence of oral cancer avoiding tobacco, limiting alcohol and increasing fruit and vegetable consumption are recommended. Recommendations for approaches to improve oral health are made for health authorities, industry, schools and health professionals and it is recommended that the Government make adequate resources available to improve oral health preventive strategies. PMID- 10800238 TI - Medical emergencies: the experience of staff of a UK dental teaching hospital. AB - OBJECTIVE: To assess the frequency of faints and other medical emergencies experienced by staff of a UK dental hospital. To investigate the training they had received in the management of medical emergencies, their perception of readiness to deal with emergencies and future training needs. SUBJECTS: All 193 clinical staff (dentists, hygienists, nurses and radiographers) of the University Dental Hospital of Manchester. DESIGN: Structured questionnaire with covering letter, reminders sent to non-responders. RESULTS: There was an 82% response. Fainting was the commonest event: other medical emergency events were experienced with an average frequency of 1.8 events per year, with the highest frequency reported by staff in oral surgery. Most expressed a need for further training: only 3% felt no need. CONCLUSIONS: Medical emergencies occur in dental hospital practice more frequently but in similar proportions to that found in general dental practice. There is a perceived need for further training among dental hospital staff in the management of medical emergencies. PMID- 10800239 TI - The effects of midazolam and flumazenil on psychomotor function and alertness in human volunteers. AB - OBJECTIVE: To investigate the effect of midazolam and flumazenil on psychomotor function and alertness in human volunteers. DESIGN: Randomised, double-blind, cross over study. METHODS: Intravenous flumazenil was administered to sedated and non-sedated healthy human volunteers, in doses typical of those used clinically to induce sedation with midazolam and for reversal with flumazenil. Subjective assessment of alertness and objective measures of psychomotor function using light reaction time and the Maddox wing were made over a 1 hour period. RESULTS: Seven males and seven females each attended four experimental sessions. Psychomotor function was impaired by midazolam but there was some individual variation to this response. All sedated subjects receiving flumazenil had significantly improved alertness and psychomotor function when compared with those subjects who received placebo. Mean alertness (P < 0.01) and light reaction time (P < 0.05) showed significant improvement and returned to baseline by 60 minutes. Stability also showed significant improvement (P < 0.05) but did not return to baseline by 60 minutes. There was no significant effect on psychomotor function or alertness when the antagonist flumazenil was administered in the absence of the agonist midazolam. CONCLUSION: An earlier discharge time based on subjective assessment of alertness is not advocated for patients whose intravenous midazolam sedation is reversed with flumazenil. PMID- 10800240 TI - Orthodontic extractions: a comparative study of inhalation sedation and general anaesthesia. AB - OBJECTIVE: To compare directly inhalation sedation and general anaesthesia in terms of treatment success rate, various aspects of morbidity and time taken, when used for patients having orthodontic extractions. DESIGN: Patients requiring orthodontic extractions were treated with either inhalation sedation or general anaesthesia. The two groups were matched for age, sex, number of teeth extracted and pre-operative anxiety. Data were collected by questionnaires. SETTING: Unit of Paediatric Dentistry at the University Dental Hospital of Manchester. SUBJECTS: All patients referred for orthodontic extractions between November 1994 and May 1996 were invited to take part in the study. Total number of patients = 101. INTERVENTIONS: Sixty-six patients commenced treatment with inhalation sedation and 35 with general anaesthesia. Routine orthodontic extractions were carried out. OUTCOME MEASURES: Treatment success rate, various aspects of morbidity and total time taken were measured and compared for the two groups. RESULTS: Treatment success rates were high for both groups. Significantly less morbidity was found to be associated with inhalation sedation and the total time taken was significantly shorter with inhalation sedation than with general anaesthesia. CONCLUSIONS: Inhalation sedation is a successful alternative to general anaesthesia for orthodontic extractions with patients experiencing less morbidity and the time taken being shorter. PMID- 10800242 TI - Clarification of Opinion 8.061 "Gifts to Physicians from Industry". Council on Ethical and Judicial Affairs of the American Medical Association. PMID- 10800241 TI - An evaluation of a training programme in primary care research for general dental practitioners. AB - AIMS: To determine whether general dental practitioners would find seminars on research methods in primary care research of interest and whether they would be stimulated to plan their own research. METHOD: Questionnaire with both open and closed questions distributed to participants at the end of the seminars. 10 seminars in locations throughout the north west of England were planned. RESULTS: 98 dentists attended the seminars of whom 81 returned questionnaires, giving a response rate of 82.6%. More than 60% found the seminars and the specially developed teaching manual very useful. However there was somewhat less enthusiasm for undertaking research projects. CONCLUSIONS: The findings suggest that some GDPs are interested in primary care research but are not keen to initiate or plan research themselves. PMID- 10800244 TI - "Pill bill" makes it easier to swallow. PMID- 10800243 TI - Nephron-sparing surgery. AB - This retrospective study examines partial nephrectomies performed over a period of approximately three years to confirm the feasibility of the procedure for suspected renal cell carcinoma. The surgeries were done primarily at a 1000-bed, university-affiliated hospital by members of a private urologic surgery practice. We examined whether recent advances in diagnosis and surgical therapy increased the viability of partial nephrectomy as an alternative to radical nephrectomy in certain carefully selected patients. Patients underwent preoperative evaluation to confirm suitability for partial nephrectomy. Surgeons used an extraperitoneal approach and mobilized the kidney after incising Gerota's fascia. Nearly one third of the procedures involved renal cooling. Because three masses turned out to be benign cysts, and one tumor was an oncocytoma, four patients with benign disease were spared unnecessary radical nephrectomy. With an average follow-up of 32 months, all patients remain free of disease and are not dependent on dialysis. Our findings are consistent with those of national studies that show partial nephrectomy does not compromise disease-free survival in properly selected patients. The success of nephron-sparing surgery in this sample opens the door for exploration of more advanced, less invasive techniques for management of renal cell carcinoma. PMID- 10800245 TI - The light at the end of the tunnel: providing meaningful patient protections. PMID- 10800246 TI - [Mineral water or tap water? A systematic analysis of the literature concerning the question of microbial safety]. AB - BACKGROUND: Based on sporadic reports of microbial contamination of mineral waters, it has been recommended that, for safety reasons, particularly immunocompromised patients should drink tap water rather than bottled mineral water. However, in terms of safety, evidence of the clinical consequences may allow a better estimate than a positive in vitro test for contamination. Therefore, we reviewed the literature on documented disease outbreaks due to contaminated mineral and tap waters. MATERIAL AND METHODS: We performed a systematic search of the literature using the database MEDLINE. In order to identify evidence relevant for Germany, we restricted our search to the years 1985-1997 (i.e. legal force of the mineral and table waters act in Germany) and the countries of Central and Northwestern Europe as well as the USA and Canada. RESULTS: Cases of contamination of tap water were documented in nearly all countries included. In 35 communications we found reports on a total of 423,000 cases of disease outbreaks due to contaminated tap water, in some cases even with lethal outcome. Main diagnosis was gastroenteritis, and main species of microorganism was crytosporidium. In contrast, there was no documented case of disease outbreak due to contaminated bottled mineral water. CONCLUSION: Tap water as well as bottled water are both supremely safe components of nutrition. The recommendation that tap water is better than mineral water, particularly for high risk patients, is not supported by the literature. PMID- 10800247 TI - [Kirlian photography: accidental or person-specific pattern?]. AB - OBJECTIVE: Is the corona electrography of Kirlian photography an accidental pattern or an individual specific discharge pattern? PATIENTS: 30 patients of the Stauferklinik Schwabisch Gmund, suffering from a great variety of diseases. STUDY DESIGN: At intervals of 10-15 min 10 Kirlian photos each of all fingers and toes were taken from each patient. Out of this material, from 8 patients 4 Kirlian photos each with evident similarity of the discharge radiographs of 3 fingers were selected. These images were scanned for mathematical procedures, and heuristic measurements of the histograms were evaluated by a genetic algorithm. RESULTS: Three pattern groups showed different heuristic measurements: accidental pattern 0.28 +/- 0.01, Kirlian pattern in general 0.3 +/- 0.02, and Kirlian pattern of the same person 0.44 +/- 0.05. CONCLUSIONS: The mathematical objectivation of the Kirlian photographs confirm the visual impression of strong similarity. Because of the lack of anatomical irregularities and since, due to new positioning, physical and chemical surface characteristics were changed in the single Kirlian photographs, the identification of the basic pattern stresses the existence of an individual discharge pattern and thus of an individual electromagnetic field. PMID- 10800248 TI - Tea tree oil: a systematic review of randomized clinical trials. AB - AIM: Tea tree oil (TTO) is immensely popular for various topical applications. In vitro studies have repeatedly demonstrated that it has antibiotic activity. This article is an attempt to systematically review the evidence from randomised clinical trials for or against effectiveness of external TTO in dermatological conditions. METHODS: Six electronic databases were searched. Methodological quality was assessed by Jadad score. Data were extracted and validated in a standardised fashion by two independent reviewers. RESULTS: Only 4 trials were located. They suggest that TTO may be effective as a treatment of acne and fungal infections. The evidence is promising but by no means compelling. The adverse effects of TTO are usually mild and transient. They mainly consist of allergic reactions. CONCLUSIONS: It is concluded that, so far, there is no compelling evidence to show that TTO is efficacious in any dermatological condition. However, in view of promising findings, TTO deserves to be investigated more closely. PMID- 10800249 TI - Clinical skills training at John A. Burns School of Medicine (JABSOM): teaching and learning the art, craft and science of clinical practice. PMID- 10800250 TI - Angioimmunoblastic T-cell lymphoma (AIL-TCL) following macrolide administration. AB - Angioimmunoblastic Lymphadenopathy with Dysproteinemia (AILD) is a rare benign reactive process which often follows exposure to certain drugs such as penicillin. Treatment with corticosteroids usually reverses the process, however there have been reports of 18% of cases evolving into non-Hodgkins lymphoma. In our case report, we present a relatively healthy woman with history of various drug hypersensitivities who developed AILD and resultant lymphoma after treatment with azithromycin. A review of the literature has failed to find reports of AILD following macrolide exposure. Clonality, not present in other forms of hyperplasia, is present in AILD and immunosuppression may account for this difference. It is difficult to say whether the drugs are simply coincidently associated or actually cause, maintain, or exacerbate clonality in AILD and facilitate malignant transformation. PMID- 10800251 TI - Postural hypertension in elderly men--the Honolulu Heart Program. AB - Postural hypertension is a condition in which the blood pressure rises abnormally during standing. The aim of this study is to determine the prevalence of postural hypertension and its association with morbidity and subsequent mortality. At the fourth examination (1991-1993) of the Honolulu Heart Program longitudinal cohort, 3741 Japanese-American men aged 71-93 were examined. Prevalence of postural hypertension in this cohort was 39%. Although there was a high prevalence of postural hypertension in this cohort, its clinical significance is questionable as there were no significant correlations with morbidity and subsequent mortality. PMID- 10800252 TI - Evaluation of late onset bipolar illness during menopause. AB - OBJECTIVE: The purpose of this paper is to review the literature on bipolar illness and to discuss its relevance to the evaluation and treatment of bipolar illness during menopause. The hypothesis is that there is a group of patients who may present with commonly reported symptoms of menopause who are in fact, suffering from an underlying bipolar illness. METHOD: The literature pertaining to gender differences in bipolar illness as well as the effect of major life events associated with the onset of bipolar illness is closely examined. RESULTS: There is enough evidence to support the hypothesis that women in particular are vulnerable to bipolar illness of the rapid cycling type. Exacerbations of a previous existing condition or late-onset bipolar illness may be associated with major stressors and life events such as that experienced during menopause. CONCLUSIONS: Gender differences in the course of bipolar illness and the greater prevalence of rapid cycling among bipolar women may in fact be a major consideration in the evaluation and treatment of symptoms during menopause. PMID- 10800253 TI - Coronary artery bypass grafting performed with or without a bypass pump: early results. AB - INTRODUCTION: Traditionally, heart bypass surgery has required stopping of the heart and the use of cardiopulmonary bypass. Numerous complications have been associated with exposure to this extracorporeal circuit. Newer techniques of local cardiac wall stabilization now enable this operation to be performed safely "Off Pump". The early clinical results of Off Pump Coronary Artery Bypass (OPCAB) will be compared to a similar group of traditional Coronary Artery Bypass Grafting (CABG) patients. METHODS: A retrospective review of 137 consecutive patients undergoing elective coronary artery bypass grafting was performed, 68 of who underwent traditional CABG and 69 of who underwent OPCAB. Inclusion criteria consisted of first time cardiac surgical procedures with an ejection fraction > or = 20%, without significant renal failure (creatinine < 2.0). RESULTS: There was no statistical difference in the age, sex, cardiac function or underlying co morbidities between those undergoing CABG and OPCAB. CABG patients had slightly more vessels bypassed than those in the OPCAB group (3.0 vs 2.6, p = 0.010). Despite similar preoperative characteristics, the OPCAB group experienced a reduction in morbidity without an increase in mortality. CONCLUSION: In similar patient populations, OPCAB was associated with significantly reduced transfusion requirements, intubation time, ICU and overall hospital lengths of stay, with no increase in mortality. Further investigation is warranted to ascertain the role of the OPCAB in the general cardiac surgical community. PMID- 10800254 TI - Use of donepezil in elderly patients with Alzheimer's disease--a Hawaii based study. AB - Donepezil (Aricept) is a reversible acetylcholinesterase inhibitor which is indicated for the treatment of mild to moderate dementia of the Alzheimer's type. We did a retrospective analysis of 41 elderly Alzheimer's subjects of different ethnic groups including a large number of Asian and Hawaiian patients. Donepezil appears to be clinically effective in patients of different ethnicities with mild to moderate Alzheimer's disease, even at advanced age. PMID- 10800256 TI - Why we need laws to protect patients from their health plan. PMID- 10800255 TI - Health education needs in Hawaii: social work, dental hygiene and nursing. AB - In this article, the need for selected health education programs in Hawaii will be discussed. Changes in the health care system, population and provider population impacting supply are identified. Health trends in Hawaii are highlighted and strategies needed to assure that Hawaii's demand for health professionals in social work, dental hygiene and nursing are suggested. PMID- 10800257 TI - Universal single payer health plan--answer to our dilemma? PMID- 10800258 TI - Magnetization transfer ratio of the spinal cord in multiple sclerosis: relationship to atrophy and neurologic disability. AB - The authors compare the spinal cord magnetization transfer ratio (MTR) of multiple sclerosis (MS) patients to healthy volunteers, relate MTR to spinal cord atrophy, and relate these and other magnetic resonance (MR) imaging parameters to disability. Sixty-five patients with MS (14 relapsing remitting [RR], 34 secondary progressive [SP], and 17 primary progressive [PP] MS), and 9 healthy volunteers were studied using MR at 1.0 T. Disability of the patients was assessed using the expanded disability status scale (EDSS). Magnetic resonance parameters were upper spinal cord MTR, number of focal spinal lesions, presence of diffuse abnormalities, and spinal cord cross-sectional area (CSA). Correlations were assessed using Spearman's rank correlation coefficient (r). Magnetization transfer ratio was higher in the controls (median, 33%; range, 30% 38%) than in patients with MS (median, 30%; range, 16-36; p < 0.05). In patients with MS EDSS correlated with spinal cord MTR, albeit weakly (r = -0.25, p < 0.05). Correlation between EDSS and spinal cord CSA was better (SRCC = -0.40, p < 0.01). No correlation was found between MTR and CSA (r = 0.1, p = 0.4). Combining MTR with spinal cord CSA improved correlation with EDSS (r = -0.46, p < 0.001), suggesting an independent correlation between disability and these 2 MR parameters. Expanded disability status scale scores were higher in patients who had diffuse spinal cord abnormality regardless of focal lesions (median, 6; range, 1.5-7.5) than in patients without diffuse abnormalities (median, 3.5; range, 0-8; p < 0.01). CSA was lower in patients with diffuse spinal cord abnormality (median, 62; range, 46-89 mm2) than in patients without diffuse abnormalities (median, 73; range, 47-89 mm2; p < 0.01). MTR was slightly lower in patients with diffuse spinal cord abnormalities (median, 29; range, 21%-33%) than in patients without diffuse abnormalities (median, 31; range, 16-36; t-test, p < 0.05). PMID- 10800259 TI - Spin-lock magnetic resonance imaging of muscle in patients with autosomal recessive limb girdle muscular dystrophy. AB - Spin-lock imaging is a new magnetic resonance imaging (MRI) technique used to reflect the microstructural integrity of muscle. The purpose of this study was to characterize spin-lock contrast (SLC) of calf muscles in limb girdle muscular dystrophy (LGMD). The calf muscles of 5 patients with LGMD and 10 healthy volunteers were imaged with an off-resonance magnetic resonance (MR) spin-lock suppression pulse. Spin-lock suppression ratios were calculated for anterior tibialis, posterior tibialis, soleus, and gastrocnemius muscles. Clinical assessments of muscle strength were compared to the spin-lock suppression ratios in the LGMD group. Strong SLC was observed in healthy muscles, with mean (+/- SD) suppression ratios ranging from 51.2% (+/- 3.6%) to 56.3% (+/- 1.3%). In diseased muscle, spin-lock signal suppression was reduced by 8%-70%, demonstrating an inverse correlation between symptom duration and suppression ratios. Spin-lock contrast in the patients with LGMD, as a reflection of tissue integrity, was best preserved in posterior tibialis, anterior tibialis, soleus, and gastrocnemius muscles in descending order. Clinical assessments did a poorer job of differentiating than SLC did and were in poor agreement with spin-lock suppression ratios. Spin-lock MRI can quantify microstructural changes in LGMD and appears to provide information not obtainable from clinical evaluations. This suggests that this noninvasive technique may be useful in evaluating the extent, progression, and response to therapy of LGMD. PMID- 10800260 TI - Utility of magnetic resonance imaging in acute intracerebral hemorrhage. AB - The authors determine whether magnetic resonance imaging (MRI) during acute hospitalization for spontaneous intracerebral hemorrhage (ICH) provides new diagnostic information. ICD-9 codes were used to identify consecutive patients with spontaneous ICH at Hermann Hospital, Houston, Texas, between January 1995 and August, 1997. Two investigators employed rigorous criteria to determine whether the MRI findings led to a specific new diagnosis. Two hundred ninety-one patients met inclusion and exclusion criteria. Sixty-seven (23%) patients underwent brain MRI during the acute hospitalization. MRI provided a new diagnosis in 15 of these 67 patients (22%). Amyloid angiopathy and vascular malformation (four each) were the most frequently identified etiologies. The yield of MRI was low in basal ganglia and thalamic hemorrhage. Two of 23 (9%) patients with deep ICH and 13 of 44 (30%) patients with lobar and infratentorial hemorrhage had etiology determined by MRI. Timing of MRI did not affect yield. PMID- 10800261 TI - Cerebrovascular findings in Takayasu disease. AB - The authors define the frequency, nature, and extent of cerebrovascular sequelae of Takayasu arteritis using functional imaging. Retrospective analysis of the cases derived from the Durban Stroke Data Bank (n = 1100) and Durban Metropolitan Vascular Surgery Database (n = 5300) consisted of evaluation by contemporary neuroimaging modalities including single positron emission computed tomography (SPECT), magnetic resonance imaging (MRI) diffusion scanning, and transcranial Doppler (TCD). Of all the patients identified with Takayasu disease (n = 142), 29 (20%) patients were identified with a primarily cerebrovascular presentation. The recent advent of modern functional imaging techniques allowed only the 10 most recent patients with a cerebrovascular presentation to be evaluated. Of these 10, 8 (80%) had normal neurologic deficit scores (Canadian neurologic score) and 9 (90%) were not disabled as determined by handicap scores (Rankin). The anatomic brain scans (9 MRI, 1 CT) were normal in 5 patients (50%). In 7 patients, transcranial Doppler sonography revealed increased velocities mainly in the anterior circulation with turbulence that was not circumscribed. Single positron emission computed tomography scanning revealed areas of hypoperfusion, mostly multiple, in all of the 7 cases investigated. The cerebral perfusion index was determined in 7 patients, with a good prognosis in 2 patients and a moderate prognosis in 5. Cerebral effects of Takayasu disease are best monitored by a combination of clinical and functional imaging such as TCD and SPECT scanning. PMID- 10800262 TI - A new pocket-sized transcranial ultrasound device (NeuroDop): comparison with standard TCD. AB - The NeuroDop is a new bedside assessment tool consisting of a continuous wave ultrasound probe attached to a stethoscope earpiece. This study was designed to compare middle cerebral artery (MCA) velocity assessment obtained with the NeuroDop versus standard transcranial Doppler (TCD). TCD technologists performed continuous wave NeuroDop studies followed by standard TCD studies on 60 subjects. Technologists recorded presence of MCA signal and estimated velocity based on NeuroDop auditory characteristics. Signal was obtained in 108 MCA vessels with the portable unit and in 112 vessels using standard TCD. For detection of patency, sensitivity was 96%, specificity 88%, positive predictive value 99%, and negative predictive value 58%. Auditorially estimated velocities from the NeuroDop strongly correlated with TCD velocity measures (r = 0.71). Categorical estimates of velocity as decreased (< 37 cm/sec), normal (37-81 cm/sec), or increased (> 81 cm/sec) demonstrated an accuracy rate of 85%. This novel stethoscope-continuous wave unit has excellent sensitivity in detecting presence of MCA patency. Moreover, MCA velocities can be characterized to a reasonable degree of accuracy based on NeuroDop auditory characteristics. The NeuroDop shows promise as a tool to rapidly assess and serially monitor presence and amplitude of MCA velocity and may help guide thrombolytic and other emergency management decisions in stroke patients. PMID- 10800263 TI - Spinal cord MRI hyperintensities in cervical spondylosis: an ischemic pathogenesis? AB - The pathophysiology of focal spinal cord MRI T2 hyperintensity (SCHI) in patients with cervical spondylosis is uncertain. This study was undertaken to determine the frequency and cause of SCHI. The authors reviewed serial cervical spine magnetic resonance imaging (MRI) reports and reviewed scans with spondylosis and cord compression or SCHI. The authors noted the location, shape, and extent of SCHI, and severity of spondylosis (expressed as a spondylosis score [SS]). The authors recorded the age and vascular risk factors for each patient. Nineteen of 273 scans (7%) with cervical spondylosis and 19 of 36 scans (53%) with cord compression had SCHI. The SCHI extended for one vertebral level from the compression in 12 patients and for three vertebral levels in 5 patients, and were distant from the compression in 2 patients. The SCHI had a focal, symmetrical, anterior spinal artery terminal zone location in 16 of 19 scans (84%). A rim isointense with normal cord separated all SCHI from the pial surface. Patients with SCHI were older (58.3 years +/- 12.8 years versus 46.8 +/- 8.1 years) (p = 0.007) and had a higher SS (5.7 +/- 2.4 versus 3.9 +/- 1.4) (p = 0.02) than patients without SCHI. The SCHI relates to the severity of cervical spondylosis. The anterior spinal artery territory location, the normal cord between SCHI and the compressive lesion, and the presence of SCHI at a distance from the compressive level all suggest an ischemic basis for SCHI. PMID- 10800264 TI - Transcranial Doppler ultrasonography: year 2000 update. AB - In this update, the main clinical applications of transcranial Doppler ultrasonography are reassessed. A specific format for technology assessment, personal experience, and an extensive review of the literature form the basis of the evaluation. The document is approved by the American Society of Neuroimaging and the Neurosonology Research Group of the World Federation of Neurology. PMID- 10800266 TI - Diffusion-weighted MRI diagnosis of pure motor stroke limited to primarily distal leg weakness. AB - Pure motor stroke (PMS) manifesting as distal weakness of a single extremity has rarely been described. The authors report a 59-year-old man with PMS who had primarily distal weakness of a single lower extremity, which to the authors' knowledge has not been previously described. Four days after onset, positive diffusion-weighted imaging confirmed that the small subcortical T2-weighted hyper enhancing and nonenhancing T1-weighted hypo-intensity noted on magnetic resonance imaging (MRI) represented an acute PMS that was the cause of the patient's weakness. Increased awareness of this rare clinical presentation of PMS coupled with the promise of imaging the PMS with diffusion-weighted MRI should lead to earlier stroke intervention in these patients. PMID- 10800265 TI - Intravascular lymphomatosis: contribution of cerebral MRI findings to diagnosis. AB - Intravascular lymphomatosis (IL) is a rare variant of non-Hodgkin's lymphoma with an unusual predilection for the central nervous system (CNS). Most cases are not diagnosed until postmortem because of variable clinical presentation and nonspecific laboratory findings. Neuroimaging findings vary widely and range from diffuse involvement of the deep white matter to infarct-like lesions. Cerebral magnetic resonance imaging (MRI) may show parenchymal and meningeal gadolinium enhancement. The authors describe brain MRI findings of linear, punctate, and patchy enhancement suggestive of CNS IL in two patients confirmed by brain biopsy/histologic studies. High index of clinical suspicion and careful interpretation of MRI (including gadolinium contrast studies) may contribute to premortem diagnosis and early intervention of this often-missed disease. PMID- 10800267 TI - Temporal lobe epilepsy presenting as panic attacks: detection of interictal hypometabolism with positron emission tomography. AB - Cerebral glucose metabolism was studied using positron emission tomography (PET) in a 13-year-old girl with a history of panic attacks that were thought to be of psychiatric origin. Positron emission tomography imaging revealed marked right temporal lobe hypometabolism and magnetic resonance imaging (MRI) detected T2 changes consistent with right mesial temporal sclerosis. Electroencephalogram (EEG) studies 3 years later confirmed a primary diagnosis of right temporal lobe epilepsy. As shown by this case and one other, PET and MRI imaging of patients with panic disorder who are thought to have epilepsy may be helpful in leading to definitive electrographic studies that confirm temporal lobe epilepsy as the primary diagnosis. PMID- 10800268 TI - Magnetic resonance and positron emission tomography changes during the clinical progression of Rasmussen encephalitis. AB - The authors describe serial positron emission tomography (PET) and magnetic resonance imaging (MRI) studies in a patient with pathologically confirmed Rasmussen Encephalitis (RE). Results of initial PET and MRI studies were normal. Subsequent studies showed involvement of the percentral and postcentral gyri and the putamen on PET, and the precentral and postcentral gyri on MRI. Coregistration of PET and MR images showed good correlation between the precentral and postcentral gyri involvement. However, subcortical involvement occurred earlier on PET than on MRI. The authors demonstrate the evolution of changes on PET and MR images in a patient with RE. Despite early pathologic confirmation of RE, there were no definite structural or functional imaging changes on PET or MRI until 3 years after symptom onset. These findings demonstrate the variability of imaging changes in RE, and the need to carefully correlate electro-physiologic and clinical findings to confirm the diagnosis of RE. PMID- 10800269 TI - Extracranial vertebral artery dissection causing cervical root lesion. AB - The authors report an unusual manifestation of extracranial vertebral artery dissection (VAD), presenting with a predominantly motor radicular manifestation. Cervical magnetic resonance imaging (MRI) revealed the intramural hematoma in the dissected vessel wall, compressing mainly the segmental motor root and, to a lesser degree, the sensory ganglion. In the digital subtraction angiography (DSA), a circumscribed narrowing of the incriminated vessel was demonstrated. Color-coded Duplex imaging (CDDI) revealed complete recanalization after a few days of anticoagulation treatment. Complete neurologic recovery was seen after 3 months. Considering the MRI data, the likely pathogenetic mechanism was compression of the nerve root by the intramural hematoma. The synopsis with similar cases in the literature points to the characteristic features, i.e., the association of neck pain with radicular motor deficit and the absence of degenerative disk disease. The respective syndrome should raise the suspicion of vertebral artery dissection, especially in young individuals. PMID- 10800270 TI - Fracture mechanics. A comparison study of torsional stress on bone. AB - Fractures that result from torsional loading of shafts in mechanical systems of nonbiologic materials generate a fracture line that forms a 45 degrees angle to an axis that is perpendicular to the direction of torsional loading on the shaft. As tension and compression are applied to these isotropic substances, the angle of fracture increases and decreases, respectively. Understanding how these forces, particularly compressive forces, generate elongation of a spiral fracture increases the ability to predict the extent of injury to bone. Fibular and metatarsal fractures are of primary importance to the podiatric physician, but any spiral fracture may be subject to torsional loading. Thus the principles stated here apply to the entire skeletal system. The purpose of this article is to provide a better understanding of the mechanics behind the causes and characteristics of fractures and to explore whether these same factors apply to the fracture mechanics of bone. PMID- 10800271 TI - Primary closure of infected diabetic foot wounds. A report of closed instillation in 30 cases. AB - Multiple surgical strategies are available for managing the infected diabetic foot at risk for amputation. The authors present their experience with the closed instillation system in the management of 30 such cases in 29 patients over a 5 year period. Data were collected from the hospital records of neuropathic patients presenting with deep-plantar-space infections or presumed acute osteomyelitis. All 29 patients were male; 57% had marginal or poor vascular supply, and 83% were nutritionally compromised or had proteinuria. At the conclusion of the study, 34% of the patients were dead, reflecting the severity of comorbid conditions found in this population. Despite the marginal healing capacity of these patients, the procedure had a 90% success rate, as defined by expeditious return to prior level of functioning and residential living situation without need for re-operation or higher-level amputation. PMID- 10800272 TI - Use of gait patterns to reveal possible disorders in the geriatric patient. AB - Changes in gait occur frequently with advancing age and are often associated with falls, loss of independence, morbidity, and even mortality. Gait changes may occur as part of the natural process of aging or may be associated with underlying pathology. Careful observation of gait can provide insight into the patient's overall state of health. This article discusses the various types of gait changes that can occur with aging and the significance of these gait abnormalities. PMID- 10800273 TI - The association of blisters with musculoskeletal injuries in male marine recruits. AB - A prospective study examining the epidemiology of blisters and, in particular, the association of blisters with subsequent injuries was conducted involving 2,130 male US Marine Corps recruits participating in initial physical training at the Marine Corps Recruit Depot in San Diego, California. From January 1993 through September 1994, recruits experienced an incidence of 2.05 blisters per 100 recruit-months. Recruits with blisters were 50% more likely to experience an additional training-related injury. Blisters, in combination with other related injuries, resulted in 159 clinic visits, 103 days of assigned light duty, and 177 lost days of training. This loss of time cost a minimum of $29,529. Extrapolating to the annual population of recruits, this represents an approximate annual expense of $690,000. Aggressive blister prevention and management in this setting has the potential to greatly reduce morbidity and fiscal costs. PMID- 10800274 TI - Proximal articular set angle. Radiographic versus intraoperative measurement. AB - The authors conducted a study to investigate the variability of measurement of the proximal articular set angle. The proximal articular set angle as measured radiographically was compared with the proximal articular set angle as measured intraoperatively. Three groups of evaluators--attending physicians, residents, and third-year podiatric medical students--measured the proximal articular set angle radiographically. The mean measurement, standard deviation, and difference between the mean radiographic measurement and the intraoperative measurement were determined for each group. The authors concluded that the only accurate proximal articular set angle measurement is the intraoperative measurement. PMID- 10800275 TI - Chondrosarcoma of the foot. AB - Chondrosarcomas have been seen clinically as aggressive tumors, with radiographic and histologic findings consistent with malignancy; however, they may have an insidious clinical course, with relatively benign radiographic and histopathologic findings. Rarely, a chondrosarcoma may present as a primary lesion of the foot. It is important to recognize these lesions as malignancies. The authors provide an overview of these neoplasms and describe a case of a low grade chondrosarcoma in a 64-year-old woman. PMID- 10800276 TI - A modified approach to the phenol and alcohol chemical partial matrixectomy. AB - The phenol and alcohol matrixectomy is a frequently used technique for treating onychocryptosis in healthy patients. This article describes a modification of this common procedure. The modified version provides a quick and easy method of removing an ingrown nail. PMID- 10800277 TI - Chondromyxoid fibroma of the fourth metatarsal. AB - Chondromyxoid fibroma is a rare benign tumor that is typically found in the metaphyseal ends of long tubular bones, such as the tibia. The authors describe a case of this neoplasm occurring in the foot. Treatment included complete resection with reconstruction using an autogenous fibular bone graft. The surgical technique and the advantages of using a fibular bone graft are discussed. PMID- 10800278 TI - Blunt dissection technique for the removal of Kaposi's sarcoma. PMID- 10800279 TI - Top priority in the new millennium: ending 400 years of racism. PMID- 10800280 TI - Assessment of serum albumin concentration as a risk factor for stroke and coronary disease in African Americans and whites. AB - Relatively high serum albumin levels compared to low levels have been associated with reduced cardiovascular mortality and coronary heart disease and stroke incidence. The nature of these associations remains undefined. Therefore, widely accepted criteria of Bradford Hill were used to assess the likelihood that the association is a causal one. These criteria were applied to the findings of published studies of serum albumin and risk of cardiovascular disease. Favoring a causal association are lack of temporal ambiguity in cohort studies, consistency of findings among studies and biologically plausible mechanisms. Not favoring a causal association are lack of specificity of the association, and inconsistency among age-, sex-groups. Further studies are needed to elucidate mechanisms for the effect of serum albumin on incidence of cardiovascular disease and death. PMID- 10800281 TI - Physical growth, sexual maturation, body image and sickle cell disease. AB - PURPOSE: This study assessed delays in physical growth and sexual maturation, self-esteem and body image in youth with homozygous sickle hemoglobin disease (HgbSS). METHOD: A consecutive sample of 30 subjects age 8 through 19 with homozygous sickle cell disease (hemoglobin SS) and a similar number of control subjects matched for age, race, gender and socioeconomic status and free of chronic illness were examined for height, weight and Tanner staging of sexual development. Subjects also completed the Body Cathexis Scale and Piers-Harris Self-Concept Scale. Assessments were with paired samples t-tests. RESULTS: The subjects with sickle cell disease had significantly lower weights and were shorter than matched control subjects. Sexual development (physical) was also delayed in the sickle cell subjects. The study failed to find significant differences for either body image or self-esteem. CONCLUSIONS: The latency age and adolescent subjects with sickle cell disease had significant delays in physical (height, weight, secondary sexual characteristics) maturation. The study failed to find significant differences in either self-esteem or body image between the two groups. Theoretical constructs from the literature were presented which questioned the belief that these expected delays in physical growth and sexual maturation have an adverse effect upon self-esteem and body image. PMID- 10800282 TI - Clinical aspects of dementia in African-American, Hispanic, and white patients. AB - This article examines the relationship between ethnicity, cognitive deficits, functional impairment, and psychiatric symptoms in patients with dementia. The data are from a cross-sectional study of patients evaluated at the Northern California Alzheimer's Disease Center (ADC). Using the ADC database of patient information, the authors compared sociodemographic and clinical variables in 187 African-American patients, 69 Hispanic patients, and 1317 white patients with Alzheimer's disease (AD), ischemic vascular dementia (IVD), and mixed dementia (AD/IVD). Multivariate analysis indicated the following results: 1. African American patients and Hispanic AD patients had lower risk of depressed mood compared to white patients; 2. African-American patients had lower risk for anxiety than whites; 3. Hispanic patients with mixed dementia had lower rates of apathy compared to whites. Future studies are needed to examine how ethnic group differences in dementia are based on the interaction of cultural differences; effects of age, education, and psychosocial variables; and biological differences in the course of dementia. PMID- 10800285 TI - Enrollment of sudden cardiac death victims into a limited cardiac autopsy study in the emergency department. AB - The lack of balanced recruitment for racial and ethnic groups in research protocols is an important ethical issue. African Americans have a history of forced participation, unethical research, and unwilling human dissection/demonstration that leads to a lack of participation in clinical research. We have embarked on a study of the pathophysiology of sudden death among African Americans which requires a limited autopsy of the heart. Our objective was to improve the recruitment rate in this study when compared with historical rates seen in clinical research and organ donation. We have screened 14 and enrolled 10 African Americans with sudden death (95% CI 0.41-0.91). By addressing the concerns of the African-American community and involving qualified input in the planning stages of the study we have been able to significantly improve the recruitment rate for this important population subgroup. PMID- 10800283 TI - Obesity differences between African-American men and women. AB - The purpose of this study was to examine the socio-demographic and behavioral differences related to obesity between African-American men and women. Obesity was defined as a body mass index of greater than 27.3 kg/m2 for women and 27.8 kg/m2 for men. Data were collected from 661 African-Americans, 418 women and 243 men, residing in wards 7 and 8 in Washington, DC through telephone interviews. Obesity was prevalent among 38.3% of the women and 20.1% of the men (p < 0.01). For women age 55 or older, annual income over $20K, having less than a high school education, and alcohol and tobacco consumption were associated with being overweight in the initial bivariate analysis (p < 0.05). For men, being 35 years or older and unemployment were significant factors associated with obesity. Our final analysis, when known dietary risk factors were adjusted, revealed that in women, obesity was associated with age, hard liquor consumption and non use of tobacco. For men, older age was a primary association. We concluded that gender, with increasing age, plays a significant role in predicting obesity, as defined by concurrent national standards. African American men 55 years of age or older are the most likely group to be overweight even after predisposing and behavioral risk factors are considered. PMID- 10800284 TI - Mutation analysis of BRCA1 gene in African-American patients with breast cancer. AB - An estimated 7% of all breast cancers and 10% of all ovarian cancers are associated with inherited mutations in BRCA1 and BRCA2 genes. The mutations of a breast cancer-susceptible gene, BRCA1, confers increased risk of breast cancer in young women. Numerous studies have reported specific mutations in the BRCA1 and BRCA2 genes in the white population. However, there are very few studies on African-American and other ethnic minority groups. The goal of this study is to identify whether African-American patients with breast cancer carry some common mutations reported in other ethnic groups and whether they carry some novel mutations. We screened hot-region mutations on exons 2, 5, 11, 16, and 20 of BRCA1 gene in 54 African-American patients with breast cancer by NIRCA and SSCP methods. Our data revealed one novel frameshift mutation (3331 insG) and three missense sequence variants (A3537G, A3667G, and C4009T) on exon 11. Each sequence change was confirmed by automatic DNA sequencing. One rare sequence variant, A3537G, has been revealed in high frequency (3/54). Our data suggested that African-American patients with breast cancer carry some unique BRCA1 gene mutations. PMID- 10800286 TI - Nasal toxicity of cocaine: a hypercoagulable effect? AB - Nasal insufflation of cocaine injures the nasal mucosa and can perforate the septum. Cocaine-induced vasoconstriction resulting in ischemia is one of the methods that may be responsible for this damage. We are determining whether cocaine also produces a hypercoagulable state that may compound factors which have been previously established to cause damage to the nasal mucosa and septum. This study uses Modified Recalcification Time (MRT), a test developed in our laboratory that has the ability to measure the overall coagulation process. Our study revealed no connection between cocaine and enhanced platelet function or monocyte-released tissue factor. The coagulation process was unaffected by the addition of the drug, so we conclude that cocaine does not cause a hypercoagulable state and cannot assist in the explanation regarding the ischemic changes of the nasal septum. PMID- 10800287 TI - Need for services in a new urban teenage clinic for men. AB - There has been excellent research in adolescent medicine in the last few years, research that is beginning to document the effectiveness of programs that can reduce high-risk behaviors. Adolescent clinics that feature medical care, condom availability, and clinic-based health education may change knowledge but do not change behavior. Linked programs that feature curriculum-based health education, conflict resolution, mentoring, and vocational and jobs programs may change knowledge and reduce risky behavior. There is little knowledge of what the pre participation opinion of the teenagers is about such services. In this descriptive study of the first 46 entrants into a new teen clinic for males, clients were surveyed about which programs were desirable. Most of the clients felt medical care and free condoms were important. Many felt job-related services were desired. One half felt a general support group and health education related to sexually transmissible infections was desirable. Education about the types of services that are potentially effective may need to be provided to adolescent males to empower them to choose appropriate services. PMID- 10800288 TI - Samoan woman, in mid-40s, examination revealed erythematous plaque of upper back and posterior neck. What is your diagnosis?. PMID- 10800289 TI - A strategic call to utilize Echinacea-garlic in flu-cold seasons. PMID- 10800290 TI - The third dimension of patient care. PMID- 10800291 TI - We can no longer ignore the rising rate of African-American suicide. PMID- 10800292 TI - Adherence to antiviral drug regimens in HIV-infected adolescent patients engaged in care in a comprehensive adolescent and young adult clinic. AB - Inconsistent use of antiviral medications for the treatment of HIV may lead to the emergence of resistant strains in HIV-infected adults. Patterns of adherence with these drug regimens in adolescents remains unknown. Identifying nonadherence in HIV-infected patients to antiviral regimens and developing corrective measures could improve patient outcomes. This study was undertaken to understand adherence in HIV-infected youths engaged in care and to reduce patterns of nonadherence. A retrospective analysis of 25 charts (78%) of HIV-infected youths (n = 32, age 13 to 21 years) were consecutively reviewed from January 1993 to May 1998. Charts were reviewed for documentation of factors previously documented to be associated with adherence: housing stability, social support, prior sexually transmitted diseases (STDs) and/or pregnancy, HIV exposure category, number of clinic visits, number of pills per day, number of medications per day, knowledge of medication schedule, age, gender, race/ethnicity, health status as revealed by CD4 count and viral load, and recorded patterns of adherence to medications and clinic appointments. Thirteen of the 18 (72%) patients who were receiving antiretroviral medication were nonadherent. Sixty-seven percent of the females and 80% of the males reported missing doses. Housing instability (p = 0.031) and/or length of treatment with antiviral medications (months of treatment) (p = 0.043) were significantly correlated with nonadherence. The stability of the adolescents' living situations was the most significant correlate of medication adherence for this population of HIV-infected youth. PMID- 10800293 TI - Racial disparities in participation in biomedical research. AB - To determine whether minority patients were less likely to participate in biomedical research, perceive positive benefits from such participation, or to recommend research participation to other patients, an observational study was conducted. Sociodemographic and survey data were collected from 5436 users of Department of Veterans Affairs (VA) Ambulatory Care, which included questions about veterans' research participation and related attitudes. Bivariate and multivariate analyses were performed to determine if there were racial differences in the outcomes of interest, controlling for relevant sociodemographic factors. Bivariate and multivariate analyses indicated that there were no racial differences in self-reported research participation, but minority veterans were more likely to perceive a positive effect of research and less likely to recommend research to other veterans. However, subgroup analyses indicated that, of those veterans having negative attitudes about research, minority and less educated veterans were disproportionately represented. In the VA system, racial differences in research participation may dissipate because many sociodemographic factors are controlled. Although we did not observe consistent racial differences in research participation or attitudes, the fact that minority veterans were disproportionately represented among the group with the most negative attitudes about research suggests that further research is necessary to fully understand the racial dynamics of research participation in the VA. PMID- 10800294 TI - Stress and coping of the African-American physician. AB - The purpose of this qualitative study was to examine the nature of work-related stress and coping experienced by African-American family physicians. Ten African American family physicians across the state of Ohio were interviewed using a standardized open-ended format. Interview data were analyzed through an "editing" technique and QSR Nud*ist, a qualitative software program. Patterns and themes common to the interviews were identified. Stressors presented by research participants included experiences with racism in medicine, doubt, and a strong desire to prove oneself in the medical environment. Distinctive coping strategies involved spirituality, kinship, and the development of strength and perseverance in the face of adversity. Responses to general questions on stress and coping indicated difficulties with the shift toward managed care and use of "time for self, away from medicine" types of coping strategies. Results underscored the importance of culture and race in stress and coping processes, and suggested that programs and policies addressing the specific pressures faced by African-American physicians in training and practice need to be developed. PMID- 10800295 TI - Crack-cocaine users as victims of physical attack. AB - This study evaluates the correlates of physical attack among people who use crack cocaine in Dayton, Ohio. Using a retrospective and prospective natural history design, data from baseline and 1-year follow-up interviews were used to calculate the prevalence of physical attack and the annual rate of physical attack suffered by 440 not-in-treatment crack-cocaine users. Logistic regression was used to determine the correlates of physical attack. The lifetime prevalence of physical attack was 63.0%; the annual rate was 36.8%. At baseline, daily crack users were more likely to report a previous attack since they began using crack (odds ratio [OR], 1.81; 95% confidence interval [CI], 1.18-2.77). Longer duration of crack use was also associated with experiencing an attack (OR, 1.09; 95% CI, 1.04 1.14). Between baseline and 12-month follow-up, the odds of men being attacked were significantly less than those for women (OR, 0.48; 95% CI, 0.23-0.99). Physical attack is widespread among crack-cocaine users, and does not vary by ethnicity. Injuries often result in the need for medical care. Over the short term, women are at increased risk. Accessible and effective drug abuse treatment is needed to diminish the harm this population suffers. PMID- 10800296 TI - Lactobacillemia: an emerging cause of infection in both the immunocompromised and the immunocompetent host. AB - The bacterium, lactobacillus, is found in the mucosal surfaces of the mouth and the gastrointestinal and genitourinary tracts. There have been increasing reports of the micro-organism being a cause of serious infection in immunocompromised individuals. This article reviews the clinical presentation, laboratory characteristics and treatment of patients with lactobacillemia. PMID- 10800297 TI - Spontaneous coronary artery dissection presenting as acute myocardial infarction. AB - Spontaneous coronary artery dissection is a rare entity being increasingly diagnosed as a cause of acute myocardial infarction, especially in cases of low cardiac risk female patients. This is one such case report of a black female patient, who suffered an acute anterior wall myocardial infarction due to an idiopathic spontaneous coronary artery dissection of the left anterior descending artery. She was treated with a thrombolytic agent in the acute phase, uneventfully. An urgent coronary angiogram demonstrated an intimal tear with a dissection of the left anterior descending artery. She survived the acute event and her subsequent hospital course was uncomplicated. Hence she was treated medically for her ischemic event and left ventricular systolic dysfunction with a favorable outcome. This case is yet another report of a survivor treated with a thrombolytic agent for the acute myocardial infarction due to spontaneous coronary artery dissection. PMID- 10800298 TI - Hemolytic disease of the newborn due to anti S antibodies. AB - We report a case of hemolytic disease in a newborn due to anti S antibodies. Baby R was born at term to an O+ mother whose antibody screen was positive for phenotype big S. Cord blood eluate revealed anti-S RBC; antigen: RBC typing for S was positive. Physical examination of baby was unremarkable. The infant's HCT was 44.2 at 6 hours of age. At 48 hours, the HCT decreased to 33.5, bilirubin peaked to 5.4, retic had peaked to 6.8. By seven days, all these values reverted to the normal, and baby has remained asymptomatic. PMID- 10800299 TI - Healthy 11-year-old girl with a 3 month history of dyspnea, non-productive cough and mild chest pain. PMID- 10800300 TI - Race and racialism in scientific research and publication in the Journal of the National Medical Association. PMID- 10800301 TI - Using non-nucleoside reverse transcriptase inhibitors in treating HIV disease: where are the therapeutic niches? PMID- 10800302 TI - [Videothoracoscopy in diagnosis and treatment of small peripheral pulmonary masses]. AB - This study was devoted to examination of efficacy of modern methods of diagnosis and surgical treatment of patients with small peripheral pulmonary masses (SPPM) with the use of videothoracoscopy. The results of examination and treatment of 126 patients with SPPM (80 patients underwent videothoracoscopy) were analysed. This study demonstrated high-quality of modern complex of clinicoradiological investigations for diagnosis of SPPM. The reliability of the complex is 91%. The necessity of immediate elective operation of the patients with SPPM is shown for verification of the process and adequate surgery. Videothoracoscopic surgery is effective method of treatment in patients with SPPM. PMID- 10800303 TI - [Selective proximal vagotomy with excision of duodenal ulcer]. AB - 257 patients with duodenal ulcer and its complications underwent selective proximal vagotomy in combination with excision of duodenal ulcer without pyloric injury. Lethality rate was 0.78%. Long-term results of the operation by Visick's classification are evaluated as excellent and good in 105 (90.5%) patients, in 9 (7.8%) as satisfactory, in 2 (1.7%) as unsatisfactory. It is emphasized that operation saving the pylorus is optimal in such patients and can be performed almost in all patients irrespective of location of the ulcer, presence and tupe of its complications. PMID- 10800304 TI - [One-row continuous suture in abdominal surgery]. AB - One-row continuous suture (OCS) was used in 3605 patients while placing anastomoses in different parts of the digestive tract, extrahepatic bile ducts and pancreas as well as suturing after gastrostomy and choledochotomy. Two techniques of OCS were used. In 526 patients OCS was used for biliary tract surgery with complications rate 1.3%. In surgery of the stomach and small intestine OCS was used in 2606 patients; the insufficiency rate of stomach and small intestine anastomoses was equal (0.04%). In surgery of colon and rectum OCS was used in 405 patients; insufficiency rate was 1.5%. In surgery of the pancreas this suture was used in 70 patients without complications after surgery. Analysis of the data shows high reliability of OCS for anastomoses in abdominal surgery and allows to recommend its wider use in clinical practice. PMID- 10800305 TI - [Comments to dispute about catarrhal appendicitis]. AB - One of the "blind-spots" in the problem of acute appendicitis is uncertainty in treatment policy for catarrhal appendicitis. Present-day pathophysiological approaches support necessity of different medical measures depending on morphological form of appendicitis. In diagnostic difficulties 186 patients underwent laparoscopy for appendicitis form specification. It is established that undestructive forms of vermiform appendix inflammation has characteristic clinico endoscopic picture presented as primary (true), and secondary (false) appendicitis. Frequency of acute appendicitis when surgical treatment is required does not exceed 10% of total amount of appendectomies. PMID- 10800306 TI - [Functional condition of pancreas after stomach resection according to Roux]. AB - Available are the results of surgical treatment of 90 patients with stenotic gastroduodenal ulcer in Burdenko Surgical Faculty Hospital of Sechenov Moscow Medical Academy between 1984 and 1985. 30 patients (study group) underwent stomach Roux-type resection. Truncal vagotomy with a stomach Bilroth-I resection was made in 20 control patients, after 20 control patients had a truncal vagotomy with pyloroplasty according to Heineke-Mikulicz, and 20 patients had selective proximal vagotomy with gastroduodenostomy by Joboulay (the third control group). Exocrine function of the pancreas was assessed by serum concentration of immunoreactive trypsin, endocrine function by fasting blood sugar, oral glucose tolerance and serum concentration of immunoreactive insulin. The authors came to the conclusion that exocrine function of the pancreas was equally damaged in patients with a Roux stomach resection, stem vagotomy with a stomach Bilroth-I resection and a stem vagotomy with pyloroplasty Heineke-Mikulicz. After selective proximal vagotomy a level of immunoreactive trypsin was normal. After a Roux stomach resection relative incompetence of basophil cells of the pancreas and long increase of insulin in the blood were observed but without influence on the glucose curve. The changes of glucose curve and level of immunoreactive insulin were similar in the control groups. PMID- 10800307 TI - [Use of autoblood and its components in orthopedics and traumatology]. AB - The results of the autoblood donation in 304 patients with complex surgical operations in orthopedics and traumatology are analyzed. The method of the intraoperation blood loss compensation with own red cells is offered. It should be noted that the red cells must be prepared 2 days and less before the operation. The use of modern methods of autoblood donation allowed to reduce 3 times the requirement in homologous blood. PMID- 10800308 TI - [Vitaflon vascular graft for portocaval shunt: experimental and clinical study]. AB - Native "Vitaflon" vascular graft made of spongeous polytetrafluoroethylene for portocaval shunt was evaluated in patients with portal hypertension. Experimental series in grey rabbits proved high biological inertness of the graft. It was used for portocaval shunt in 13 patients. The follow-up from 1 to 32 months demonstrated successful results in 12 cases. PMID- 10800309 TI - [Treatment of multiple ribs fractures]. AB - Of 267 patients with multiple fractures of the ribs, 69 patients had mobile costal valves. As a rule, the presence of the floatable chest wall site, is a cause of increasing respiratory failure ending in lethal outcome in 40 to 80% of cases. Indications for combined conservative therapy and its scope are specified and efficiency of different methods of restoration the chest wall frame is evaluated. The technique of pericostal ligature fixation of the mobile costal valves to the thorax splint offered by the authors was improved. The performance of this manipulation may be essentially easier thanks to the use of the device for passing ligature around the ribs and breast bone, which was developed by the authors and patented in Russian Federation. Clinical application of the developed technique allows rather quickly to control respiratory failure. The lethality tell 6 times in comparison with patients who were treated with the use of the other methods of the floatable costal valves fixation. PMID- 10800310 TI - [Sepsis. Evaluation of views, necessity of terminology unification and diagnostic criteria]. AB - Critical analysis of modern overview devoted to general infectious-inflammatory process and materials of original research (690 patients) provides a basis for discussion of diagnostic criteria of and new terminology. Wide introduction of researches on classification, criteria of diagnosis and terminology offered by North American conference (R. Bone et al., 1992) into practice is recommended. The given definitions better reflect the essence of homeostatic disorders in septic patients, allow to carry out early change of the program of treatment, to standardize approaches to diagnosis, monitoring and therapy and enable search in epidemiology of sepsis, choice of the best ways of scientific investigation. PMID- 10800311 TI - [Plastic and reconstructive operations in purulent surgery and traumatology]. AB - The authors propose a complex surgical treatment of extensive purulent wounds and tissue defects based on radical debridement of the pyonecrotic focus, primary and early reconstructive operations. New views on operation "debridement of a purulent wound" were formulated: it is considered as an initial element of the reconstructive surgery. 597 patients with extensive purulent wounds and tissue defects underwent plastic and reconstructive operations with good and satisfactory results in 96.6% of the cases. PMID- 10800312 TI - [Surgical treatment of fecal incontinence in children]. AB - The experience of the faeces incontinence treatment in 295 children (age ranged from 1 to 15 years) is presented. Several original operations for surgical correction of the sphincter ani asthenia are proposed. The authors think that radical correction of faeces incontinence in children is necessary as early as at the age 6 to 7 years that is at the beginning of social activity. Efficient use of the above methods of the rectum closing apparatus creation allowed to achieve good results in 77.3-85.7% of the children. PMID- 10800313 TI - [Treatment of erectile dysfunction in cases of venous and corporovenous insufficiency of phallus]. AB - A total of 230 patients of different age with impaired venous drainage of penis cavernous bodies were examined. Test with intracavernous injection of papaverin, dopplerography of the vessels and cavernosometry were employed. To treat venous and corporovenous insufficiency, it is suggested to make a resection of the deep dorsal vein, ligation of the superficial and circular veins with suturing tunica albuginea. In negative result of the surgery viagra in a done 50 (100) mg is recommended or penile implants. PMID- 10800314 TI - [Treatment of chronic arterial insufficiency of legs with midocalm in elderly]. PMID- 10800315 TI - [Abdominal foreign bodies of traumatic genesis]. PMID- 10800316 TI - [Use of sodium hypochlorite solution for prophylaxis of postoperative purulent wounds complications]. PMID- 10800317 TI - [Stalked gastrostomy in cicatricial stricture of esophagus]. PMID- 10800318 TI - [Current classification of peritonitis and system estimating severity of patient's illness]. PMID- 10800319 TI - [Problem of informed consent in medicine]. PMID- 10800320 TI - [Gastrointestinal carcinoids]. PMID- 10800321 TI - [Local hemostasis in surgery of liver and spleen injuries]. PMID- 10800322 TI - [A method for determining the tonus of the stomach wall in patients with duodenal peptic ulcer complicated by organic pyloroduodenal stenosis]. AB - Method of gastric wall tone determination before the operation in patients with gastroduodenal ulcer disease, complicated by pyloroduodenal stenosis (PDS) was elaborated. There were examined 86 patients with sub- and 62 with decompensated PDS using method of electrogastrography and of the intragastric pressure registration--using method of the opened catheter. Three degrees of the gastric motor function disorder were delineated. Tactics of treatment of such patients was elaborated. PMID- 10800323 TI - [Methods to prevent generalized peritonitis in peptic ulcer surgery]. AB - Different variants of omentoduodeno- and omentogastroplasty were applied in 69 patients during performance of gastric resection, perforative ulcer excision for the peritonitis dissemination prophylaxis. In 18 patients was used the polychlorvinyl drain, newly constructed by the authors. In 124 patients gastric resection according to Haberer with an oblique gastroduodenoanastomosis formation, the Treitz ligament dissection and duodenojejunal flexure descending were performed for the peritonitis dissemination prophylaxis. PMID- 10800324 TI - [The prevention and treatment of Curling's ulcers in children]. AB - Experience of prophylaxis and treatment of stress ulcers of Curling and hemorrhage from them in 250 children with burn, aged from 5 months to 14 years, was analyzed. Complex of prophylactic measures, intensive antishock therapy and the actions algorithm in the hemorrhage occurrence from Curling's ulcers was elaborated, the experience of the membrane plasmapheresis application in children with severe and extremely severe burn was summarized. PMID- 10800325 TI - [An approach to the right hepatic duct in treating high damage and cicatricial stricture of the bile ducts]. AB - There was proposed an operative access to the duct via the lateral end of transverse hepatic fissura, basing on the results of studying of the right hepatic duct anatomy. The method performed includes the right hepatic lobe mobilization, its fibrous capsule transsection along the transverse fissura in the direction toward right posterior sectorial duct, longitudinal transsection of the duct lateral wall along 1-2 cm with subsequent formation of laterolateral hepaticojejunoanastomosis. Method was tested in 9 patients with high damage and cicatricial stricture of biliary ducts, the result was positive. PMID- 10800327 TI - [The function of the coagulation hemostatic and fibrinolytic processes in the postoperative period in patients with complicated chronic pancreatitis]. AB - Intravenous infusion of modified amino acid cocktail (AC), based of the "Aminosyn PF" composition with addition of glutamine, methyonine and selenium was applied in the complex of treatment of patients with complicated chronic pancreatitis. After AC infusion during 3 days after the operation the coagulation indexes restoration was noted while after conventional treatment in these terms the signs of thrombohemorrhagic syndrome were registered. PMID- 10800326 TI - [Current approaches to the surgical treatment of liver cirrhosis complications]. AB - Complex hemo- and lymphodynamic investigations were performed in 298 patients with liver cirrhosis and syndrome of portal hypertension. Tactics of surgical intervention for hemorrhage from varicose veins of esophagus and stomach was elaborated. Indications for the shunting and disconnection operation performance were established. New methods of the disconnecting operation performance were proposed. Main roentgenoanatomic types of the ductus thoracicus (DT), morphology and types of intrahepatic lymphatic drainage were depicted. Indications for the operations, stipulating the DT draining, extraperitonization of liver and peritoneovenous shunting, were substantiated. PMID- 10800328 TI - [Intralobar pulmonary sequestration]. AB - For the period of 21 years the intralobar pulmonary sequestration was revealed in 6 patients aging from 11 to 47 years old. In all the observations of the intralobar sequestration the only affection of lower pulmonary lobes was noted. In 4 patients the symptomless course of the disease was present, while in 2 it was manifested by the recurrent pneumonia. The correct diagnosis before the operation was established in one patient only. All the patients were operated on, in 2 two aberrant vessels to the sequestrated pulmonary region were revealed and in all the patients--bronchogenic cyst in the affected region. PMID- 10800329 TI - [The characteristics of operating on and postoperative management in patients with pulmonary tuberculosis and diabetes mellitus]. AB - Peculiarities of the operations performance and the postoperative period conduction in 128 patients with pulmonary tuberculosis and diabetes mellitus (DM) were enlightened. After the operation in majority of patients it is expedient on the background of diet and insulin therapy to continue conventional chemotherapy. In patients with severe DM, focal pulmonary changes, chronic alcoholism and those to whom the preliminary chemotherapy was not conducted, it is mandatory to prescribe an intensive chemotherapy. For correction of the carbohydrate metabolism disorders it is expedient to conduct electrophoresis of 6% solution of sodium salicilate and 5000 U of heparin according to the elaborated method. PMID- 10800330 TI - [The surgical treatment of toxic goiter in middle-aged and elderly patients]. AB - Of 128 patients, ageing more than 60 years, polynodular toxic goiter was diagnosed in 84, diffuse toxic goiter--in 34, toxic adenoma--in 10. The nodules in thyroid gland (TG) were revealed in 94 patients, cancer--in 3. There were following indications for the operation: the lack of the thyreostatic therapy efficacy, the malignant-like nodules presence, compression of cervical and mediastinal organs. After the euthyreotic state achievement all the patients were operated on using endotracheal narcosis. Subtotal resection of TG was performed in 91 patients, thyroidectomy--in 27, hemithyroidectomy--in 10. The frequency of specific complications (n. recurrence damage, bleeding, hypoparathyroidism, the wound purulent complication) was 4.7%. PMID- 10800331 TI - [The complications of wound healing after an operation for ventral hernia and their prevention]. AB - For abdominal hernia 982 patients were operated. The complications had occurred in 18.6% of patients due to chronic inflammation and the presence of "sleeping" infection in the hiatal gates margins. Application of the elaborated prophylaxis complex of measures had permitted to lower the frequency of the wound healing complications down to 9%. PMID- 10800332 TI - [The clinical characteristics, diagnosis and treatment of soft-tissue suppurative inflammatory diseases in patients with narcotic dependence]. AB - Comparative analysis of clinical course and the diagnostic methods of purulent inflammatory disease of soft tissues was performed in 20 patients with drug abuse (the 1st group) and in 20 patients, who are keeping the normal way of living (2nd group). In 16 (80%) of patients of the 1st group the anaerobic non-clostridial infection (ANI) was diagnosed. The factors causing the ANI occurrence, were determined, the programme of treatment was introduced. In the 1st group 3 patients died. The duration of the stationary course in the 1st group was 18 days and in the 2nd one--3.8 days. PMID- 10800333 TI - [The use of color duplex scanning in the diagnosis of vascular testicular disorders in surgical diseases of children]. AB - Results of investigation of 37 boys, ageing from 3 to 15 years, were analyzed, in whom surgical pathology of processus vaginalis peritonealis using the duplex scanning method with coloured duplex mapping was revealed. The proposed complex of treatment of children, assumes maximal preservation of the organ during the operation and the blood supply restoration after it. PMID- 10800334 TI - [Knee joint instability: the current view of the problem]. AB - Modern instrumental methods of diagnosis of the knee joint (KJ) fracture and of the damaged structures identification are presented. The proposed modern methods of operative treatment of the KJ posttraumatic instability, which are applied in the clinic, are recommended for broad usage. PMID- 10800335 TI - [The errors and complications in corrective operations for scoliosis]. AB - The analysis of mistakes and complications after 4377 correcting operations, performed in 2719 patients for severe scoliosis, was done. The working classification and on her background computer programme of disease diagnosis, prophylaxis and treatment were proposed. Mistakes of preoperative period were mainly tactical, while during the operation--technical, in early postoperative period--clinical and in the late postoperative period--orthopedic. The presence of mistakes lowered significantly the treatment efficacy, they had become the cause of mortality. PMID- 10800336 TI - [The functional treatment of transcondylar and epicondylar fractures of the humerus in children]. AB - Functional treatment of 275 children with epicondylar and transcondylar fracture of shoulder bone with II-IV degree dislocation was done. The conventional method of functional treatment was applied using dissected in articulation ulnaris plaster bandage, the Blaunt bandage and the elaborated method using the external fixation apparatuses--hinge-rotational apparatus (HRA). The highest efficacy of functional treatment using HRA was noted. PMID- 10800337 TI - [The mechanisms for the decrease in gastric motor-evacuatory function after performing a vagotomy]. AB - In experiment on white rats in was established that after the truncal vagotomy performance in the stress gastric ulcer the lowering of the serotonin and histamine blood content was noted as well as the disorder of ratio of their concentration in gastric wall. Insufficiency of serotonin causes the disorder of gastric motor-evacuation function after the operation. Administration in postoperative period of serotonin adipinate had promoted the raising of serotonin concentration in blood and gastric tissues, histamine level lowering, enhancement of serotonin/histamine ratio. PMID- 10800338 TI - [The role of the intestines in the pathogenesis of acute pancreatitis: oxygen extraction and bacteria translocation in rats]. AB - In experiment on 40 male rats of the Wistar line there was investigated the oxygen extraction (O2) and the bacteria translocation in 30 min, 2, 6, 12 and 24 hours after the stimulating operation (control group) and simulation of the ductal-hypertensive form of an acute experimental pancreatitis (AEP)--the main group. The data obtained witness the general extraction of O2 raising by 222% (P < 0.001) at average and lowering of its intestinal extraction in 30 min and 2 hours (by 33 and 31% at average, P < 0.05) and its raising in 24 hours by 138% (P < 0.001) at average also. These disorders had correlated with velocity of the intestinal E. coli reproduction and with the bacteria translocation on the way intestine-->mesenterial lymph nodes in 6 hours after the disease occurrence with preservation of the lymph nodes and the liver barrier function in the AEP duration over 24 hours. The data obtained had permitted to substantiate the necessity of the remedial measures complex application, directed on the frequency of purulent complications lowering. PMID- 10800339 TI - [Compression biliodigestive anastomosis: the morphological characteristics of its dynamics of formation]. AB - The method of the compressive biliodigestive anastomosis formation, using implant and the apparatus for its performance, was elaborated. An experimental investigation on 30 rabbits was done. An optimal strength of compression while doing the tissues connection was 0.002-0.004 N/m2. Minimal traumaticity, preservation of sufficient blood flow of the tissues connected, delineation of the healing zone from the intestinal contents and bile, the absence of chronic inflammation in anastomotic tissues are the advantages of compressive biliodigestive anastomosis, promoting early restoration of the intestinal and gallbladder wall, the formation of functionally full value anastomosis, which is not stenotizes cicatricially. PMID- 10800340 TI - [The morphogenesis and pathogenesis of a metadiaphyseal infarct of the long bones in different types of ischemia]. AB - In experiment on the animal's bones the various kinds of ischemia was simulated (obturational, compressive, traumatic) it was established that the central mechanism of the bone infarction pathogenesis is irreversible blockade of microcirculatory bed by cellular aggregates and microthrombi. The most extended and constant infarction occurs in embolization of the main nourishing artery and in osteotomy of diaphysis with the bone marrow intersection and the periosteum detachment on the ends of fragments. PMID- 10800341 TI - [The treatment procedure in acute pancreatitis]. PMID- 10800342 TI - [The meaning of the term multiple trauma]. PMID- 10800343 TI - [A self-fixing drainage device]. PMID- 10800344 TI - [The use of hemo-carboperfusion in the combined treatment of patients with suppurative-septic diseases of the hepatobiliary organs]. PMID- 10800345 TI - [Reconstructive gastric resection for exclusion for duodenal ulcer after performing a Finney pyloroplasty and subdiaphragmatic anterior truncal vagotomy]. PMID- 10800346 TI - [An organized hematoma of the peribladder space following herniotomy for a femoral hernia]. PMID- 10800347 TI - Second opinions. PMID- 10800348 TI - He that hath clean hands. PMID- 10800349 TI - Emergency medical services: scene of injury to emergency department--key issues for spinal cord injuries. PMID- 10800350 TI - Acute management of patients with spinal cord injury. PMID- 10800351 TI - Rehabilitation and wellness after spinal cord injury. PMID- 10800352 TI - The public vocational rehabilitation program. A resource for individuals with spinal cord injury. PMID- 10800353 TI - Assistive technology for people with spinal cord injuries: what physicians need to know. PMID- 10800354 TI - Accessing medical care with a spinal cord injury. PMID- 10800355 TI - Overview of Rhode Island tobacco laws. PMID- 10800356 TI - "TAKE CARE Rhode Island" coalition. PMID- 10800357 TI - Benign schwannoma. PMID- 10800358 TI - Hospitalizations for spinal cord injuries, 1994-1998. PMID- 10800359 TI - [Quality control in ambulatory major surgery]. PMID- 10800360 TI - [Systematic recording of perioperative events associated with anesthesia as quality indicator in ambulatory anesthesia]. AB - OBJECTIVES: To analyze the quality of several anesthetic techniques used for major outpatient surgery in our hospital, by quantifying for each the relative risk (RR) of adverse events during anesthesia and in the postoperative period. PATIENTS AND METHODS: One thousand seventeen patients who underwent surgery between 18 May 1998 and 23 October 1998 were studied retrospectively. RESULTS: The mean age of the patients was 52.27 +/- 24.65 yr; 44.18% were ASA I, 40.56% were ASA II, 14.56% ASA III and 0.67% ASA IV. Mean time of surgery was 33 +/- 16.49 min and mean recovery time until discharge was 77.3 +/- 93.4 min. Admission was necessary for 0.6% of the patients and re-admission for 0.3%. General anesthesia was used with 19%, anesthetic monitoring with 17%, regional anesthesia (including peribulbar) with 46% and local anesthesia plus sedation with 16.6%. In 95% of the cases, no adverse events occurred during anesthesia; in 94.8% no such events occurred during the early recovery period. During surgery and postoperative recovery, intradural anesthesia was associated with significantly greater RR of adverse events in comparison with general anesthesia (6.6 and 2.2 respectively) and in comparison with monitored anesthesia (7.2 and 3.3). No differences in RR were found between general anesthesia and monitored anesthesia. Problems were slight to moderate in severity and mainly related to nausea and vomiting (2%). CONCLUSIONS: Recording perioperative events permits evaluation of the quality of anesthesic procedures. Intradural anesthesia is associated with more complications. PMID- 10800361 TI - [Intraoperative control of mean arterial pressure and heart rate with alfentanyl with fuzzy logic]. AB - OBJECTIVES: To describe a fuzzy logic controller that adjust alfentanil infusion during surgery based on changes in mean arterial pressure (MAP) and heart rate (HR). MATERIAL AND METHODS: We designed a fuzzy logic controller using if ... then ... conditions written in C language, to be executed by a 486 PC with a 66 Mhz CPU. The controller was used with eight ASA I-II patients undergoing gynecological surgery under anesthesia with propofol, alfentanil and ventilated with oxygen/air. MAP and HR were input every three minutes, after which the controller generated an infusion based on those figures. We performed a statistical study of alfentanil consumption time until extubation and time of hemodynamic stability. Relative error of MAP was calculated. RESULTS: The controller was used for a total of 373 min with the eight patients. MAP was 15% below the desired level for 2.14% (18 min) of that time and was 15% over the desired level for 5.6% (21 min) of the time. MAP held steady within the range of stability for the remaining 92.26% (334 min) of the time the controller was used. The relative error of MAP was 7.8 +/- 1.5%. Mean time until extubation was 7 min and 2 s. CONCLUSIONS: We believe the controller can be used to automate taks executed by experts. The controller was useful for stabilizing HR and MAP. PMID- 10800362 TI - [AIDS and anesthesia]. AB - AIDS concerns anaesthetic practice for various reasons. First, this syndrome can affect different organs that have anaesthetic implications. Second, drugs usually taken by the patient can interact with anaesthetic agents. And last, the risk of infection for health workers must be taken into account, as well as the procedure to follow after accidental injures. PMID- 10800363 TI - [Abdominal compartmental syndrome in non-injured patients]. AB - Abdominal compartment syndrome (ACS) is characterized by increased intraabdominal pressure and a set of secondary pathophysiological changes in the abdominal. ACS has reappeared in the literature recently in relation to the surgical concept to damage control, applied particularity in contexts of severe abdominal injury polytraumatized patients. We report two cases of ACS that appeared after scheduled abdominal surgery: one after repair of a large eventration and the other in the context of septic shock due to fecaloid peritonitis. Both patients died of multisystem organ failure in spite of surgical decompression. We wish to emphasize that ACS can appear in contexts other than surgery for damage control, and we stress the need to measure intravesical pressure as a reflection of intraabdominal pressure, particularly in certain high risk patients in the postoperative recovery ward. Finally, we review the pathophysiology of ACS and its management, which is based on early treatment to prevent multisystem organ failure with an associated high risk of death. PMID- 10800364 TI - [Tension pneumoperitoneum as complication in a case of perioperative barotrauma]. AB - Tension pneumoperitoneum is rare complication that causes severe changes in respiratory and cardiocirculatory function. We report the case of an 84 year old woman who developed tension pneumoperitoneum soon after cholecystectomy by subcostal laparotomy. Tension pneumoperitoneum presented with barotrauma, bilateral pneumothorax, extensive subcutaneous and pneumomediastinum and was accompanied by severe worsening of hemodynamic variables and gas exchange. After confirming the diagnosis by emergency CT scan, the first therapeutic action was decompression of the peritoneal. This fast, simple maneuver was followed by rapid hemodynamic and respiratory improvement. Tension pneumoperitoneum similar to the case we report has been described as a complication of mechanical ventilation and barotrauma; however, the possibility that the clinical picture might be caused by perforation of a hollow viscera has given rise to a variety of suggested therapeutic alternatives, including exploratory laparotomy, which has sometimes later proven to have been unnecessary. In the light of the case we report and the literature, we suggest a therapeutic sequence starting with emergency abdominal decompression and to fulfill two objectives: a) to remove the threat to life, and b) to limit the practice of emergency exploratory laparotomy to cases in which that technique is truly indicated. PMID- 10800365 TI - [Epidural anesthesia for cesarean section in a patient with Marfan syndrome]. PMID- 10800366 TI - [The inhalator Ventolin produces artifacts in the anesthetic gas analyzer]. PMID- 10800367 TI - [Anesthesia in a case of laparoscopic repair of Morgagni's hernia]. PMID- 10800368 TI - [Administration and monitoring of inhaled nitric oxide]. PMID- 10800369 TI - [Capnography and hypovolemic shock]. PMID- 10800370 TI - [Intraoperative transesophageal echocardiography]. PMID- 10800371 TI - Science and religion: friends or foes? AB - There is a widespread assumption that science and religion are incompatible with each other; it may be acceptable for a scientist to practise religion, but--it is believed--this should be seen as despite, not because of his or her scientific profession and knowledge. Such an assumption is wrong. Examination of conflicts between science and religion (such as those about evolution) show that they always arise through misinterpretation or tenacious presuppositions. Unfortunately protagonists (both anti-religion and anti-science) receive more publicity than conciliators. This paper reviews some of the background and development of arguments over creation and evolution and shows that there is no need to choose between a rigorous scientific approach and an avowedly religious acknowledgement of God as creator and sustainer. PMID- 10800372 TI - Science and religion: rack or featherbed--the uncomfortable supremacy of science PMID- 10800373 TI - The greying world: a challenge for the twenty-first century. AB - As we approach the next millennium, an unprecedented rapid increase of total and relative numbers of older persons in both the developed and the developing world is being observed. The total aged population (aged more than 60 years) world-wide will rise from 605 million in the year 2000 to 1.2 billion in 2025. Population ageing is due to two factors: increased life expectancy and decreasing fertility rates. Public health interventions are major contributor to both. A significant consequence of ageing is the shift from communicable to non-communicable diseases. The World Health Organization (WHO) is addressing the multiple challenges posed by population ageing through integrated activities that include consolidation of databases, capacity building (research and training), advocacy and policy development. These are all approached through WHO perspectives on ageing and health: life-course, health promotion, socio-economic, community based, intergenerational, cultural, gender and ethical. PMID- 10800374 TI - Historical eclipses and the earth's rotation AB - Ancient eclipse observations are proving of considerable value in modern geophysics. These seemingly crude observations enable variations in the length of the day--produced by tides and other mechanisms--to be investigated in some detail over more than two millennia. The main attraction of the early data is the long time scale which they cover. They reveal long-term trends which cannot be discerned from the much more accurate telescopic observations. Useful historical records of eclipses originate from only four early cultures: Babylon, China, Europe and the Arab dominions. The observations fall into two main categories: timings of both solar and lunar eclipses and qualitative descriptions of total or near-total solar eclipses. Eclipse timings are due to astronomers, but chroniclers have contributed many detailed accounts of large solar eclipses. Analysis of these various observations reveals that over the past 2,500 years the average rate of lengthening of the day has been 1.7 milliseconds per century, significantly less than the tidal figure of 2.3. PMID- 10800375 TI - The Cambrian explosion and the slow burning fuse AB - The rapid appearance of animal phyla in the fossil record during the 'Cambrian explosion' ca 543 Myr ago marks the most conspicuous turning point in earth history. This 'explosion' was preceded by a 'slow burning fuse', from the start of the prokaryote fossil record at ca 3450 Myr BP to endosymbiotic assembly of the eukaryote cell between ca 2,700 and 1,000 Myr. Research is beginning to put these events into their environmental context. Very long periods of environmental stability are suggested by the carbon isotopic and palaeoclimatic record prior to ca 1,000 Myr. Such stasis may have nurtured endosymbioses to the point at which eukaryotic organization and sexual reproduction became embedded in the genome. This steady state world was chaotically disrupted in the prelude to the Cambrian explosion. Strontium, sulphur and carbon isotopes attained maximal values during this time, and the latter show chaotic oscillations coincident with flips between extreme, low latitude glaciations and possible supergreenhouse conditions. These chaotic bifurcations may have been caused by tectonically driven increases in nutrient flux to the oceans and/or by the impact of multicellularity on the carbon cycle. Whatever the cause, high rates of biotic turnover during these times of stress could have radically redirected and/or accelarated the path of evolution towards new animal body plans. PMID- 10800376 TI - Mathematical modelling of species formation PMID- 10800377 TI - Progress and challenges in geochronology AB - Rapidly improving technologies have provided increasingly precise and widely applicable means for measuring geologic time. Whilst the ability to resolve age continues to improve, the absolute accuracy will remain limited by sources of systematic error that loom ever larger in comparison. The decay constants that characterize radioactive decay rates, and govern the relationship between isotopic data and their corresponding radioisotopic ages, are inexactly known. In consequence, the accuracy of some of the most precise dating methods, such as the 40Ar/39Ar technique, may be an order of magnitude or more worse than their precision. Scientific inquiries requiring comparison of ages determined by different radioisotopic systems, such as thermochronology or early Solar System evolution, are most impacted by these accuracy-limiting effects. Amelioration of the condition is straightforward (though not necessarily easy), but requires shifting research priorities and more interdisciplinary interaction. PMID- 10800378 TI - [Biologically degradable implants in trauma surgery]. PMID- 10800379 TI - [Biodegradable implants in fracture fixation: state of the art]. AB - Operative fracture repair in trauma surgery is currently performed using metal implants. These metal implants often are removed during a second, retrieval operation. Biodegradable fracture fixation devices have been used clinically since the late seventies. Most bioresorbable implants are manufactured from polymers. The polylactides, polyglycolides and co-polymers slowly degrade into small components that are excreted from the human body via natural pathways and removal operations after fracture surgery are not necessary. Due to the limited mechanical properties, the polymer screws and pins are mostly used in the treatment of non weight-baring simple fractures of the ankle, elbow, hand and foot. In view of the progressing technical developments, new materials will be developed and tested for clinical use in the coming decades. PMID- 10800380 TI - [Biomechanics of interlocked nailing in humeral shaft fractures. Comparison of 2 nail systems and the effect of interfragmentary compression with the unreamed humeral nail]. AB - In this biomechanical study the implanted Unreamed Humeral Nail (UHN) has been tested concerning bending and torsional stiffnesses. In literature other intramedullary implants have been criticized for insufficient rotatory stability especially in transverse and short oblique fractures of the humeral shaft. This study examined, whether the implanted UHN, as well as the UHN implanted with interfragmentary compression through a specific compression device, is able to augment torsional stiffness significantly. To evaluate bending and torsional stiffnesses, the UHN has been compared biomechanically to the Russell-Taylor humeral nail (RT) in paired mid-shaft osteotomized cadaveric humeri. Identic paired comparison has been performed with the UHN without and UHN with interfragmentary compression. In anterior-posterior, as well as medio-lateral direction stiffness under four-point-bending is significantly higher in stabilizing with the RT. Under torsional loading with moments of 4 Nm, 6 Nm and 8 Nm the UHN reached more than the double torsional stiffness. The RT, which is only dynamically interlocked, owns a high initial "play" between bolts and nail itself. Through additional interfragmentary compression stiffness of the UHN under four-point-bending in anterior-posterior, as well as medio-lateral direction augments significantly. Also under torsional loading with moments of 4 Nm, 6 Nm und 8 Nm torsional stiffness increases with interfragmentary compression significantly. In comparison to other biomechanical studies of different authorship it is clear, that this statically interlocked intramedullary nailing of the humeral shaft is superior to non-statically interlocked types of nailing concerning their stabilizing potency in torsion and serves especially for fracture types, which are critically under rotation, as transverse or short oblique humeral shaft fractures. PMID- 10800381 TI - [Treatment of complex elbow injuries by joint-spanning articulated fixator]. AB - Biomechanical investigations in eight specimen elbow joints to test articulated external fixation showed correct extension-flexion joint position between 0 and 120 degrees. The central axis through the radial and ulnar condyles varies less than 3 degrees in total. X-ray cinematography showed correct articulation in all cases without subluxation. In six patients with persistent subluxation and complex fracture of the proximal ulna external fixation with an additional articulation was placed. All patients began with physiotherapy on the 3rd day after operation, the range of joint motion was 0-30-95 degrees in all cases, after 6 weeks 0-30-110 degrees. Additional articulated external fixation at the elbow joint can be an alternative treatment to long-term transfixation of the elbow in cases of complex injury. PMID- 10800382 TI - [Indication and results of ulna shortening osteotomy in ulnocarpal wrist joint pain]. AB - The ulnar impaction syndrome is proven to be a common source of ulnar sided wrist pain. Ulna-shortening osteotomy represents a successful therapy for this kind of problem, both congenital or posttraumatic positive ulnar variance. Positive variance resulting from a distal radius fracture needs correct dorsal and radial angulation of the radius. In case of congenital positive variance arthroscopic debridement for decompression of the TFCC should be performed first. The adequate correction of the length is the major problem. Disorders of the distal radioulnar joint may result due to overcorrection. Oblique osteotomy using 7-hole-plates is our preferred treatment. PMID- 10800383 TI - [Lesions of peripheral structures of cadaver fingers caused by Kirschner wires]. AB - Fractures of the hand skeleton are common injuries. These are or are associated with phalangeal fractures. Kirschner wires can be used for osteosynthesis, but can cause lesions of peripheral finger structures. The aim of our study was to record the incidence of lesions in peripheral structures of the fingers caused by retrograde crossed inserted Kirschner wires of 1.0, 1.25, 1.4 and 1.6 mm diameter into the middle and proximal phalanges of the long fingers and the proximal phalanx of the thumb of 120 fresh cadaver hands. We found an increase of insertion attempt frequency and an increasing number of lesions with increasing Kirschner wire diameter. Because of these findings Kirschner wires of 1.0 or 1.25 mm diameter should be preferred to prevent the peripheral functional finger structures. PMID- 10800384 TI - [Microangiographic evaluation of vascular supply of the medial collateral ligament of the knee joint]. AB - Because the vascularization of ligaments is an important factor in ligament healing we wanted to examine the vascularization status of the medial collateral ligament of the knee. The type of vascularization was examined by microangiographie of 21 postmortem preparations. We could observe a vascularization from the proximal end of the ligament by one or two major vessels in all cases. These vessels were mostly localized in the ventral third of the ligament. From the proximal to the distal part of the ligament we found a decreasing density of blood vessels. We could not observe vessel free ligament areas. Due to this vascularization status we would advise the differentiation of two ligament areas respecting the following therapy. Because of the good vascularization in the proximal part of the ligament we can accept good conditions concerning complete healing after a non-operative treatment and an operative therapy by suture or refixation. In the case of interligamentous and distal ruptures operative reconstruction by sutures often in difficult and a conservative therapy leads to a high rate of poor results. Therefore we advise fibrin bonding in these ligament areas if a surgical procedure is indicated because of the high risk of vessel strangulation after suture. PMID- 10800385 TI - [Cast immobilization versus vacuum stabilizing system. Early functional results after osteosynthesis of ankle joint fractures]. AB - In a prospective randomized trial the early functional results after immobilisation in a cast were compared to those after using a vacuum stabilizing system. The vacuum stabilizing system Vacoped offers equivalent stability compared to a plaster cast. In contrast to the cast the Vacoped can be removed for body care and physical therapy. Additionally the range of motion for dorsal flexion/extention in the upper ankle joint can be adjusted. From 9/1996 to 7/1997 there were 40 patients included in the study with an operated ankle fracture as monotrauma. Six weeks postoperatively the patients with cast treatment showed significantly higher functional deficits for the upper ankle joint (20%), the lower ankle joint (40%) and muscle atrophy (2.1 cm side difference) than the group with the vacuum stabilizing system (upper ankle joint 15%, lower ankle joint 25%, 1.4 cm muscle atrophy). Five patients out of the group with the vacuum system were already at work three weeks postoperatively. Three months postoperatively the functional results for both groups were approximating. The vacuum stabilizing system Vacoped offers better early functional results than conventional cast treatment after osteosynthesis of ankle fractures. Because of the increased patient comfort and the early ability for physical therapy the vacuum stabilizing system is preferable to cast treatment. PMID- 10800386 TI - [Dystonia following peripheral trauma: clinical findings and diagnostic criteria]. AB - Dystonia is a rare neurological complication after peripheral trauma. Incidence and pathophysiology of post-traumatic dystonia are not known. Predisposing factors are sympathetic reflex dystrophia, pre-existing movement disorders or a family history of movement disorders. The main diagnostic goal is to exclude other causes of secondary dystonia. Objective criteria for posttraumatic dystonia are not established, and therefore differentiation from psychogenic dystonia frequently remains difficult. Careful psychiatric examination is obligatory. Clinical criteria are consistency of the symptoms over time and the presence of symptoms compatible with organic dystonia. Polygraphic EMG examinations provide objective correlates of the movement disorder, but exact EMG criteria for the diagnosis of dystonia have yet to be established. PMID- 10800387 TI - [Specific features of accidents, injuries and surgical care in the elderly]. PMID- 10800388 TI - [Luxatio antebrachii ulnaris: a very rare form of elbow dislocation]. AB - We report about a 61 years old patient, who suffered a rare ulnar dislocation of the elbow after being arrested by the police. PMID- 10800389 TI - [Bilateral hip ankylosis caused by heterotopic ossification after isolated severe craniocerebral trauma]. AB - A 36-year old man suffered an isolated head injury with a fracture of the skull, epidural and subdural hematomas as well as brain contusion (Abbreviated Injury Scale (AIS): 5 points). The hematomas were evacuated by craniotomy. Because of high intracranial pressure (ICP) a barbiturate coma was necessary. Additionally, the patient demonstrated acute lung injury (ARDS) due to pneumonia 8 days after trauma. The patient recovered slowly and was transferred to neurorehabilitation on day 57 after injury. During the following 12 months the patient developed a significant deficit of hip motion (extension/flexion: right: 0 degree/10 degrees/20 degrees, left: 0 degree/10 degrees/30 degrees; external/internal rotation: right and left: < 5 degrees/0 degree/< 5 degrees; abduction/adduction: right: 10 degrees/0 degree/25 degrees, left: 10 degrees/0 degree/10 degrees). X rays and CT-scanning revealed severe heterotopic ossification (HO) of both hips with ancylosis (Brooker type IV). Resection of HO was carried out in a two stage procedure using Smith-Petersen approach. The prophylaxis for recurrence of HO included preoperative single-dose radiation (8 Gy) and postoperative treatment with indomethacin (150 mg per day). The patient revealed 15 (left hip) and 12 (right hip) months after surgery the following range of hip movement: extension/flexion: right: 5 degrees/0 degree/90 degrees, left: 5 degrees/0 degree/100 degrees; external/internal rotation: right: 20 degrees/0 degree/30 degrees, left: 20 degrees/0 degree/20 degrees; abduction/adduction: right: 30 degrees/0 degree/40 degrees, left: 30 degrees/0 degree/40 degrees. No recurrence of HO was observed in x-ray. The patient is able to work in his profession as farmer. PMID- 10800390 TI - [Occult fracture of ossified Achilles tendon without associated rupture of the tendon]. AB - Ossification of the Achilles tendon is the result of a traumatic, hypoxic injury of the tendon. The usually asymptomatic ossification has a clinical importance only in case of a new trauma. We report about a patient with an isolated fracture of the ossification without an accompanying rupture of the Achilles tendon. Because of the radiologically "occult" fracture the diagnosis could only be verified by MRI. With a conservative treatment without resection of the ossification we could reach painless recovery. PMID- 10800391 TI - [Problems of informed consent]. PMID- 10800392 TI - [Reconstructive surgery of the cruciate ligament. How to achieve optimal results? The Indianapolis experience]. PMID- 10800393 TI - Mesothelial proliferations. An increasing morphologic spectrum. PMID- 10800394 TI - Fine-needle aspiration cytology in the diagnosis of lymphoma. The next step. PMID- 10800395 TI - The new era of automated immunoassay. AB - Use of immunoassays and other ligand-binding assays in clinical diagnosis has increased dramatically during the last several years. Despite impressive technical advances, "mass production" of these assays in a routine laboratory still presents many difficulties. This review of ligand-binding assay technology highlights some recent developments, emphasizing challenges and possible solutions for cost-effective patient care. PMID- 10800396 TI - Lymphohistiocytoid mesothelioma. An often misdiagnosed variant of sarcomatoid malignant mesothelioma. AB - Three cases of lympho-histiocytoid mesothelioma, a rare variant of pleural sarcomatoid malignant mesothelioma, are described. Histologically, the neoplasms were characterized by a diffuse discohesive proliferation of atypical histiocytoid cells intermixed with a marked lymphocytic and lesser plasmacytic infiltrate. One case initially was misdiagnosed as a ganglioneuroma, a second case was misinterpreted as malignant lymphoma, and a third case was sent in consultation with the differential diagnosis of inflammatory pseudotumor vs mesothelioma. Immunohistochemical studies showed strong and generalized expression of cytokeratins and vimentin by the neoplastic histiocytoid cells in all 3 cases. Two cases were positive for calretinin, one of which also was positive for HBME-1, thrombomodulin, and LeuM1. None of the cases stained with the epithelial glycoprotein markers carcinoembryonic antigen, B72.3, and Ber-EP4, or the blood group antigen, BG-8. The immunophenotype of the lymphoplasmacytic infiltrate revealed predominantly reactive, mature T cells, with fewer polytypic plasma cells, histiocytes, and B cells. In lymphohistiocytoid mesothelioma, as in the usual examples of sarcomatoid mesothelioma, the demonstration of cytokeratin expression by the neoplastic cells is the most useful diagnostic finding that allows exclusion of other neoplasms with which this entity may be confused. PMID- 10800397 TI - Pulmonary capillary hemangiomatosis-like foci. An autopsy study of 8 cases. AB - Pulmonary capillary hemangiomatosis (PCH) typically occurs in young patients who have signs and symptoms of pulmonary hypertension. It commonly is misdiagnosed in life as pulmonary veno-occlusive disease, and the correct diagnosis usually is not made until autopsy. Autopsy records, including reports, gross photographs, histologic slides, clinical histories, and radiographic images, were reviewed to identify cases with morphologic changes characteristic of PCH. The previous case reports describe PCH as a diffuse process throughout both lung fields. All patients were symptomatic, and most died of the disease. This article details 8 cases of PCH-like foci that were incidental findings at autopsy in which the patients did not have symptoms of pulmonary hypertension nor did PCH contribute in any way to death. This is the first case series that describes pathologic changes of PCH occurring in this setting, and we hope to provide more interest in PCH and its natural history. PMID- 10800398 TI - h-Caldesmon as a specific marker for smooth muscle tumors. Comparison with other smooth muscle markers in bone tumors. AB - Caldesmon is a protein widely distributed in smooth and non-smooth muscle cells and is thought to regulate cellular contraction. Its isoform, high-molecular weight caldesmon (h-CD), was demonstrated to be specific for smooth muscle cells and smooth muscle tumors of the soft tissue and to never be expressed in myofibroblasts. We performed an immunohistochemical study to examine h-CD expression in the following bone tumors: conventional and non-conventional osteosarcoma, 13; malignant fibrous histiocytoma of bone, 5; giant cell tumors of bone, 5; chondroblastoma, 3; metastatic leiomyosarcoma, 2; and rhabdomyosarcoma, 1. Frequent immunoreactivity for muscle actin (alpha-smooth muscle actin or muscle-specific actin) was seen in 11 of 13 osteosarcomas and all other tumors, whereas h-CD was expressed intensely only in 2 leiomyosarcomas. h-CD is considered a specific and useful marker to distinguish smooth muscle tumor from bone tumors with myoid differentiation. PMID- 10800399 TI - Immunohistochemical determination of HER-2/neu expression in invasive breast carcinoma. AB - Numerous methods exist for HER-2/neu assessment; however, technical and interpretive standardization is virtually absent. We evaluated 2 commercially available antibodies on routinely fixed paraffin-embedded tissue sections to establish our own guidelines. Thirty-three cases of infiltrating breast carcinoma were evaluated simultaneously with monoclonal and polyclonal antibodies. Only membranous staining, no matter how focal, was considered positive. An additional 32 tumors were studied subsequently using only the polyclonal antibody. Of all carcinomas, 13.0% showed immunohistochemical evidence of HER-2/neu overexpression. High-grade tumors were more often positive. There was no HER 2/neu gene expression in the benign epithelium that generally was present in the tissue section or in any of the well-differentiated tumors tested. The polyclonal antibody proved more sensitive than the monoclonal antibody. While true cytoplasmic staining was present occasionally, it did not create substantial difficulty in interpretation. The polyclonal antibody cost substantially less than the monoclonal antibody. Fluorescence in situ hybridization assay for HER 2/neu gene amplification performed on 32 of 65 cases showed concordant results in 31 cases. The immunohistochemical assay for HER-2/neu gene overexpression, using our methods, is accurate, economic, and easily integrated into the laboratory. PMID- 10800400 TI - Sensitivity of HER-2/neu antibodies in archival tissue samples of invasive breast carcinomas. Correlation with oncogene amplification in 160 cases. AB - Overexpression and amplification of the HER-2 oncogene in patients with breast cancer has correlated with early onset of metastasis, resistance to hormonal therapy and some forms of chemotherapy, and shortened survival. Therefore, evaluation of this putative prognostic or predictive factor seems critical. Because different antibodies are used for the detection of the 185-kd HER-2 oncoprotein, we studied the sensitivity of 3 frequently used antibodies. Immunohistochemistry results were correlated with gene amplification level as assessed by fluorescence in situ hybridization. Protein overexpression was found in 17.2% and 12.5% of cases using antibodies against the external (TAB250) and internal (CB11) domains of the protein, respectively, and in 38.0% of cases using a rabbit polyclonal antibody. Fluorescence in situ hybridization was successful in all 160 tumors, and amplification was found in 37 tumors (23.1%). The monoclonal antibody TAB250 had the lowest misclassification rate, 9.6% (sensitivity, 67%; specificity, 97.5%). PMID- 10800401 TI - Uroplakin III is a highly specific and moderately sensitive immunohistochemical marker for primary and metastatic urothelial carcinomas. AB - Uroplakins are specific differentiation products of terminally differentiated superficial urothelial cells. We tested the value of a new commercially available monoclonal antibody against uroplakin III (clone AU 1) as a paraffin-reactive immunohistochemical marker for primary and metastatic urothelial carcinomas. The study cases included 67 urothelial carcinomas of the urinary tract (35 primary tumors, 32 metastases) and 318 nonurothelial carcinomas, as well as 5 benign Brenner tumors and 2 transitional cell carcinomas of the ovaries. Uroplakin III was detected in 21 (60%) of the primary urothelial carcinomas and 17 (53%) of the metastases, resulting in an overall sensitivity of 0.57. The studied Brenner tumors also were immunoreactive for uroplakin III. All other studied carcinomas were consistently uroplakin III-negative (specificity 1.00). We found the new monoclonal antibody AU 1 against uroplakin III to be a highly specific paraffin reactive immunohistochemical marker for urothelial tumors with a moderate sensitivity for the identification of primary and metastatic urothelial carcinomas. PMID- 10800402 TI - Diagnosis and subclassification of primary and recurrent lymphoma. The usefulness and limitations of combined fine-needle aspiration cytomorphology and flow cytometry. AB - The primary diagnosis of non-Hodgkin lymphoma/leukemia by fine-needle aspiration (FNA) is still controversial and relatively underused. We evaluated our FNA experience with lymphomas using the revised European-American classification of lymphoid neoplasms to determine the reliability of FNA when combined with flow cytometry in the diagnosis of lymphoma, the types of diagnoses made, and the limitations of this technique. Slides and reports from all lymph node and extranodal FNAs performed during the period January 1, 1993, to December 31, 1998, with a diagnosis of lymphoma or benign lymphoid process were reviewed. There were 290 aspirates from 275 patients. These included 158 cases of lymphoma, of which 86 (54.4%) were primary and 72 (45.6%) were recurrent. There were 44 aspirates suggestive of lymphoma and 81 benign/reactive diagnoses. With diagnoses suggestive of lymphoma considered as positive for lymphoma, levels of diagnostic sensitivity and specificity were 95% and 85%, respectively. Specificity was 100% when only definitive diagnoses of lymphoma were considered. Clearly, FNA and immunophenotyping by flow cytometry are complementary and obviate a more invasive open biopsy for many patients with lymphadenopathy. PMID- 10800403 TI - Pathologic findings for bacille Calmette-Guerin infections in immunocompetent and immunocompromised patients. AB - The pathologic findings from biopsy specimens from 9 patients with postvaccination bacille Calmette-Guerin (BCG) infection are presented. The patients were vaccinated with BCG during the first 2 days of life. Four patients had normal immunity and 5 patients were immunocompromised. The pathologic findings in both groups were different. Biopsy specimens from patients with normal immunity showed multiple epithelioid granulomas and Langhans giant cells with or without suppuration. Caseous necrosis was minimal. Ziehl-Neelsen stain for acid-fast bacilli showed a few bacilli in 2 cases and was negative in the remaining 2 cases. Biopsy specimens from the second group of patients, who were immunosuppressed, consisted mainly of skin and subcutaneous tissue. These revealed diffuse infiltrates of histiocytes with plump nuclei and abundant "dirty" grayish cytoplasm, which was full of numerous acid-fast bacilli. The clinical course for the 2 groups also was different. Patients with normal immunity generally recover completely, spontaneously or after excision of the suppurative lymph node and usually do not require antibiotic chemotherapy. In immunosuppressed patients, disseminated BCG infection, which may prove fatal, may develop. These patients should receive a full course of antituberculous chemotherapy and, in addition, treatment of the underlying immunologic disorder. PMID- 10800404 TI - Poor predictive ability of urinalysis and microscopic examination to detect urinary tract infection. AB - Results of urinalysis, particularly the leukocyte esterase and nitrite tests, often are used to determine whether treatment is needed or a culture will be performed in cases of suspected urinary tract infection. However, there is disagreement over the quality of urinalysis as a screening test for urinary tract infections. Final urine culture results (n = 225) were obtained from the clinical microbiology laboratory. Stepwise binary logistic regression was used to derive a model using presence of infection as determined by culture as the dependent variable and urinalysis results as independent variables. A second set of data (n = 128) then was obtained to test the model. Statistical significance and the ability to predict infection based on urinalysis results were determined. Results indicated a lack of sensitivity for leukocyte esterase, nitrite, and presence of bacteria in the microscopic examination as indicators of urinary tract infection. PMID- 10800405 TI - Evaluation of a PCR probe capture assay for the detection of Toxoplasma gondii. Incorporation of uracil N-glycosylase for contamination control. AB - Toxoplasma gondii is a cyst-forming parasite of clinical relevance in humans primarily because of the neurologic abnormalities it can cause. In some clinical circumstances, it is desirable to detect the pathogen directly. We modified a commercially available Toxoplasma polymerase chain reaction (PCR) probe capture assay by incorporating uracil N-glycosylase (UNG) to prevent carryover amplicon contamination. In addition, UNG inactivation and DNA denaturation were accomplished chemically to simplify the DNA hybridization to the capture probe. The incorporation of UNG effectively eliminated carryover contamination; the probe capture assay showed a log increase in detection sensitivity compared with standard agarose gel electrophoresis. To assess sensitivity and possible inhibition of amplification, different sample types were spiked with Toxoplasma organisms. After DNA extraction and PCR amplification, a sensitivity of 2 tachyzoites for the assay was determined in buffered saline, cerebrospinal fluid (CSF), serum, and amniotic fluid; 20 tachyzoites for whole blood; and 200 tachyzoites for brain tissue. An additional 20 human serum and CSF samples submitted for Toxoplasma serologic testing were run by the PCR method. Of these, only an IgM-positive CSF sample was PCR positive. The Toxoplasma PCR probe capture assay showed good sensitivity and was not substantially inhibited by several different clinically relevant samples. PMID- 10800406 TI - Effects of unfractionated heparin, low-molecular-weight heparin, and heparinoid on thromboelastographic assay of blood coagulation. AB - Thromboelastography (TEG) has been used increasingly as an intraoperative hemostasis monitoring device. Low-molecular-weight heparins are given increasingly to reduce the development of antibodies against the heparin-platelet factor 4 complex, and heparinoids are given to patients who have developed the antibody. We studied the effect of unfractionated heparin, a low-molecular-weight heparin (enoxaparin sodium [Lovenox]), and a heparinoid (danaparoid sodium [Orgaran]) on blood clotting assayed with TEG (TEG clotting) in vitro and the efficacy of protamine sulfate and heparinase for reversing the effect. Heparin, enoxaparin, and danaparoid all caused a dose-dependent inhibition of TEG clotting of normal blood. Concentrations of enoxaparin and danaparoid that totally inhibited TEG clotting only minimally prolonged the activated partial thromboplastin time. While inhibition of TEG clotting by heparin and enoxaparin was reversed by protamine sulfate and heparinase, inhibition by danaparoid was reversed only by heparinase. Abnormal TEG clotting was observed in patients receiving enoxaparin whose plasma level of the drug was more than 0.1 antiXa U/mL. However, the degree of TEG abnormality did not always coincide with plasma levels of the drug. PMID- 10800408 TI - Establishing reference ranges for PT and aPTT times. PMID- 10800407 TI - Transfusion-associated graft-vs-host disease. A fatal case caused by blood from an unrelated HLA homozygous donor. AB - Transfusion-associated graft-vs-host disease (TA-GVHD) is a rare complication of transfusion. We report fatal TA-GVHD in a 63-year-old coronary artery bypass patient of European descent after an RBC transfusion from an unrelated donor. The patient had mild lymphocytopenia and received 2 80-mg doses of methylprednisolone and 7 units of RBCs. On day 14 after the transfusion, he had fever, elevated liver enzyme levels, and a macular rash. Pancytopenia and bone marrow aplasia developed. On day 26, he had a massive gastrointestinal hemorrhage and died. At autopsy, histopathologic findings of the skin, liver, bone marrow, and gastrointestinal tract were consistent with TA-GVHD. One donor of the transfused RBCs (3 days old at transfusion) had a 1-way HLA match with the patient. A method using multiplex polymerase chain reaction is presented. This patient with TA-GVHD and mild immune suppression suggests that blood component irradiation guidelines may need to be reevaluated. PMID- 10800409 TI - Are the new reagents really more sensitive? PMID- 10800410 TI - Problems associated with increased sensitivity. PMID- 10800411 TI - The past and future of public health practice. PMID- 10800412 TI - Heralding a change: field action reports. PMID- 10800413 TI - Public health practice and the journal. PMID- 10800414 TI - Human rights: the foundation of public health practice. PMID- 10800415 TI - The potential and limitations of data from population-based state cancer registries. AB - Cancer incidence varies markedly among states because of population heterogeneity regarding risk, genetic, and demographic factors. Population-based cancer registries are essential to monitoring cancer trends and control. The Centers for Disease Control and Prevention and the North American Association of Central Cancer Registries, through the National Program of Cancer Registries, are helping state registries generate more and better data nationwide. The National Program of Cancer Registries has supported the enhancement of 36 registries and the creation of 13 new registries in 45 states, 3 territories, and the District of Columbia, providing national standards for completeness, timeliness, and quality; financial support; and technical assistance. Users must be aware of diverse issues that influence collection and interpretation of cancer registry data, such as multiple cancer diagnoses, duplicate reports, reporting delays, misclassification of race/ethnicity, and pitfalls in estimations of cancer incidence rates. Attention to these issues and intense use of the available data for cancer surveillance will enable maximum societal benefit from the emerging network of population-based state cancer registries. PMID- 10800416 TI - Helping the urban poor stay with antiretroviral HIV drug therapy. AB - Recent studies have documented dramatic decreases in opportunistic infections, hospitalizations, and mortality among HIV-infected persons, owing primarily to the advent of highly active antiretroviral medications. Unfortunately, not all segments of the population living with HIV benefit equally from treatment. In San Francisco, only about 30% of the HIV-infected urban poor take combination highly active antiretroviral medications, as compared with 88% of HIV-infected gay men. Practitioners who care for the urban poor are reluctant to prescribe these medications, fearing inadequate or inconsistent adherence to the complicated medical regimen. Persons typically must take 2 to 15 pills at a time, 2 to 3 times a day. Some of the medications require refrigeration, which may not be available to the homeless poor. Most homeless persons do not have food available to them on a consistent schedule. Therefore, they may have difficulty adhering to instructions to take medications only on an empty stomach or with food. Lack of a safe place to store medications may be an issue for some. In addition, many urban poor live with drug, alcohol, or mental health problems, which can interfere with taking medications as prescribed. Inconsistent adherence to medication regimens has serious consequences. Patients do not benefit fully from treatments, and they will become resistant to the medications in their regimen as well as to other medications in the same classes as those in their regimen. Development of resistance has implications for the broader public health, because inadvertent transmission of multidrug-resistant strains of HIV has been demonstrated. Concern that the urban poor will not adhere to highly active antiretroviral medication regimens has led to debate on the role of clinicians and public health officials in determining who can comply with these regimens. Rather than define the characteristics that would predict adherence to these regimens, the San Francisco Department of Public Health created a program to support adherence among those who may have the greatest difficulty complying with complicated highly active antiretroviral medication regimens. The program, dubbed the Action Point Adherence Project, was conceived through a community planning process in preparation for a city-wide summit on HIV/AIDS that took place in January 1998. Action Point is funded by the city and the county of San Francisco. Now in its 10th month, the program continues to show promising evidence of improving clients' biological and social indicators. PMID- 10800417 TI - Don Quixote, Machiavelli, and Robin Hood: public health practice, past and present. AB - Since the mid-19th century, when the first formal health departments were established in the United States, commissioners, directors, and secretaries of public health have functioned as senior members of the staffs of public executives, mayors, governors, and presidents. They have provided important political, managerial, and scientific leadership to agencies of government that have played increasingly important roles in national life, from the sanitary revolution of the 19th century to the prevention of HIV/AIDS and the control of tobacco use today. Although public health officials come from a variety of backgrounds and oversee agencies of varied size and composition, there are philosophical themes that describe and define the commonality of their work. These themes are captured metaphorically by 3 celebrated figures: Don Quixote, Machiavelli, and Robin Hood. By turns, the public health official functions as a determined idealist (Don Quixote), a cunning political strategist (Machiavelli), and an agent who redistributes resources from the wealthier sectors of society to the less well off (Robin Hood.) All 3 personae are important, but, it is argued, Robin Hood is the most endangered. PMID- 10800418 TI - Antagonism and accommodation: interpreting the relationship between public health and medicine in the United States during the 20th century. AB - Throughout the course of the 20th century, many observers have noted important tensions and antipathies between public health and medicine. At the same time, reformers have often called for better engagement and collaboration between the 2 fields. This article examines the history of the relationship between medicine and public health to examine how they developed as separate and often conflicting professions. The historical character of this relationship can be understood only in the context of institutional developments in professional education, the rise of the biomedical model of disease, and the epidemiologic transition from infectious disease to the predominance of systemic chronic diseases. Many problems in the contemporary burden of disease pose opportunities for effective collaborations between population-based and clinical interventions. A stronger alliance between public health and medicine through accommodation to a reductionist biomedicine, however, threatens to subvert public health's historical commitment to understanding and addressing the social roots of disease. PMID- 10800419 TI - Preparing currently employed public health nurses for changes in the health system. AB - OBJECTIVES: This article describes a core public health nursing curriculum, part of a larger project designed to identify the skills needed by practicing public health workers if they are to successfully fill roles in the current and emerging public health system. METHODS: Two focus groups of key informants, representing state and local public health nursing practice, public health nursing education, organizations interested in public health and nursing education, federal agencies, and academia, synthesized material from multiple sources and outlined the key content for a continuing education curriculum appropriate to the current public health nursing workforce. RESULTS: The skills identified as most needed were those required for analyzing data, practicing epidemiology, measuring health status and organizational change, connecting people to organizations, bringing about change in organizations, building strength in diversity, conducting population-based intervention, building coalitions, strengthening environmental health, developing interdisciplinary teams, developing and advocating policy, evaluating programs, and devising approaches to quality improvement. CONCLUSIONS: Collaboration between public health nursing practice and education and partnerships with other public health agencies will be essential for public health nurses to achieve the required skills to enhance public health infrastructure. PMID- 10800420 TI - Public health advocacy: process and product. AB - OBJECTIVES: In this article the author describes public health advocacy and proposes a conceptual framework for understanding how it works. METHODS: The proposed framework incorporates the image of an assembly line. The public health advocacy assembly line produces changes in societal resource allocation that are necessary for optimizing public health. The framework involves 3 main stages: information, strategy, and action. These stages are conceptually sequential but, in practice, simultaneous. The work at each stage is continually adjusted according to circumstances at the other stages. RESULTS: The framework has practical implications; for example, public health advocacy teams need members with complementary skills in distinct roles. Potential applications are illustrated via two public health advocacy efforts. CONCLUSIONS: The framework may be useful in assessing staffing and funding needs for public health advocacy endeavors, explaining common problems in these endeavors and suggesting solutions, and guiding decisions concerning effort allocation. Application of the framework to a variety of public health advocacy endeavors will clarify its strengths and weaknesses. PMID- 10800421 TI - Effectiveness of food fortification in the United States: the case of pellagra. AB - OBJECTIVES: We evaluated the possible role of niacin fortification of the US food supply and other concurrent influences in eliminating the nutritional deficiency disease pellagra. METHODS: We traced chronological changes in pellagra mortality and morbidity and compared them with the development of federal regulations, state laws, and other national activities pertaining to the fortification of cereal-grain products with niacin and other B vitamins. We also compared these changes with other concurrent changes that would have affected pellagra mortality or morbidity. RESULTS: The results show the difficulty of evaluating the effectiveness of a single public health initiative such as food fortification without controlled experimental trials. Nonetheless, the results provide support for the belief that food fortification played a significant role in the elimination of pellagra in the United States. CONCLUSIONS: Food fortification that is designed to restore amounts of nutrients lost through grain milling was an effective tool in preventing pellagra, a classical nutritional deficiency disease, during the 1930s and 1940s, when food availability and variety were considerably less than are currently found in the United States. PMID- 10800422 TI - Impact of vaccine financing on vaccinations delivered by health department clinics. AB - OBJECTIVES: This study measured the number of childhood vaccinations delivered at health department clinics (HDCs) before and after changes in vaccine financing in 1994, and it assessed the impact of changes in financing on HDC operations. METHODS: We measured the number of vaccination doses administered annually at all 57 HDCs in New York State between 1991 and 1996, before and after the financing changes. Interviews of HDC personnel assessed the impact of financing changes. A secondary study measured trends in Pennsylvania and California. RESULTS: HDC vaccinations for preschool children in New York State declined slightly prior to the financing changes (6%-8% between 1991 and 1993) but declined markedly thereafter (53%-56% between 1993 and 1996). According to nearly two thirds of New York State's HDCs, the primary cause for this decline was the vaccine-financing changes. HDC vaccinations for preschool children in Pennsylvania declined by 12% between 1991 and 1993 and by 56% between 1993 and 1997. HDC vaccinations for polio-containing vaccines in California declined by 31% between 1993 and 1997. CONCLUSIONS: Substantially fewer vaccinations have been administered at HDCs since changes in vaccine financing, thereby keeping preschool children in their primary care medical homes. PMID- 10800423 TI - Effect of the tobacco price support program on cigarette consumption in the United States: an updated model. AB - OBJECTIVES: This study evaluated the direct effect of the tobacco price support program on domestic cigarette consumption. METHODS: We developed an economic model of demand and supply of US tobacco to estimate how much the price support program increases the price of tobacco. We calculated the resultant increase in cigarette prices from the change in the tobacco price and the quantity of domestic tobacco contained in US cigarettes. We then assessed the reduction in cigarette consumption attributable to the price support program by applying the estimated increase in the cigarette price to assumed price elasticities of demand for cigarettes. RESULTS: We estimated that the tobacco price support program increased the price of tobacco leaf by $0.36 per pound. This higher tobacco price translates to a $0.01 increase in the price of a pack of cigarettes and an estimated 0.21% reduction in cigarette consumption. CONCLUSION: Because the tobacco price support program increases the price of cigarettes minimally, its potential health benefit is likely to be small. The adverse political effect of the tobacco program might substantially outweigh the potential direct benefit of the program on cigarette consumption. PMID- 10800424 TI - The effect of a structured smoking cessation program, independent of exposure to existing interventions. AB - OBJECTIVES: This study assessed the effectiveness of a smoking cessation program for women in public health clinics, controlling for reported exposures to 4 common intervention components (provider advice, booklet, video segment, posters) among smokers in the control group. METHODS: After a baseline control period, 10 pair-matched clinics were randomly assigned to study groups. A total of 1042 smokers in the combined baseline and control groups and 454 smokers in the intervention group completed a preintervention questionnaire and a postintervention telephone interview 5 to 8 weeks later. Eight smoking outcomes, including quitting, were analyzed for the effect of reported exposure to intervention components, experimental program, and clinic service. RESULTS: Greater exposure to intervention components, being in the experimental program, and being seen in prenatal clinics independently improved smoking outcomes. CONCLUSIONS: The number of interventions reported by smokers in the control group ranged from none to 4 and varied across clinic services. The experimental program we tested produced better outcomes than the minimal smoking cessation interventions already existing in the control clinics, after we controlled for whether smokers were or were not exposed to these interventions. PMID- 10800425 TI - The impact of workplace smoking ordinances in California on smoking cessation. AB - OBJECTIVES: The effect of local workplace smoking laws in California was assessed to determine whether such laws increase smoking cessation. METHODS: Workplace smoking ordinance data from 1990 were appended to 1990 California Tobacco Survey data from 4680 adult indoor workers who were current cigarette smokers or reported smoking in the 6 months before the survey. Ordinance effects on cigarette smoking and worksite policy were estimated by using multiple logistic regression controlling for sociodemographic variables. RESULTS: Smokers who worked in localities with a strong workplace ordinance (compared with no workplace ordinance) were more likely to report the existence of a worksite smoking policy (odds ratio [OR] = 1.6; 95% confidence interval [CI] = 1.2, 2.2) and to report quitting smoking in the prior 6 months (OR = 1.5; 95% CI = 1.1, 1.7). In communities with strong ordinances, an estimated 26.4% of smokers quit smoking within 6 months of the survey and were abstinent at the time of the survey, compared with an estimated 19.1% in communities with no ordinance. CONCLUSIONS: Workplace smoking ordinances increased smoking cessation among employed smokers, indicating that these laws may benefit smokers as well as nonsmokers. PMID- 10800426 TI - Failure to defend a successful state tobacco control program: policy lessons from Florida. AB - OBJECTIVES: This investigation sought to define policy and political factors related to the undermining of Florida's successful Tobacco Pilot Program in 1999. METHODS: Data were gathered from interviews with public health lobbyists, tobacco control advocates, and state officials; news reports; and public documents. RESULTS: As a result of a recent legal settlement with Florida, the tobacco industry agreed to fund a youth anti-smoking pilot program. The program combined community-based interventions and advertisements. In less than 1 year, the teen smoking prevalence rate dropped from 23.3% to 20.9%. The program also enjoyed high public visibility and strong public support. Nevertheless, in 1999, the state legislature cut the program's funding from $70.5 million to $38.7 million, and the Bush administration dismantled the program's administrative structure. Voluntary health agencies failed to publicly hold specific legislators and the governor responsible for the cuts. CONCLUSIONS: The legislature and administration succeeded in dismantling this highly visible and successful tobacco control program because pro-health forces limited their activities to behind-the-scenes lobbying and were unwilling to confront the politicians who made these decisions in a public forum. PMID- 10800428 TI - Uses of Behavioral Risk Factor Surveillance System data, 1993-1997. AB - OBJECTIVES: The purpose of this study was to document and describe Behavioral Risk Factor Surveillance System (BRFSS) data use patterns, benefits, and barriers from 1993 to 1997. METHODS: Data use information was gathered via a Medline database search and a telephone survey of BRFSS program directors (n = 54). RESULTS: The database search uncovered 109 BRFSS-based reports. Program directors indicated that BRFSS data frequently were used to support health policies regarding diabetes, physical activity, and smoking. Frequent data use barriers included insufficient special population data, insufficient city- or county specific data, and insufficient staff. CONCLUSIONS: Use of BRFSS data, which aid several state health activities, increased from 1993 to 1997. PMID- 10800427 TI - Cigarettes and suicide: a prospective study of 50,000 men. AB - OBJECTIVES: This study examined the relation between smoking and suicide, controlling for various confounders. METHODS: More than 50,000 predominantly White, middle-aged and elderly male health professionals were followed up prospectively with biennial questionnaires from 1986 through 1994. The primary end point was suicide. Characteristics controlled for included age, marital status, body mass index, physical activity, alcohol intake, coffee consumption, and history of cancer. RESULTS: Eighty-two members of the cohort committed suicide during the 8-year follow-up period. In age-adjusted analyses with never smokers as the comparison group, the relative risk of suicide was 1.4 (95% confidence interval [CI] = 0.8, 2.3) among former smokers, 2.6 (95% CI = 0.9, 7.5) for light smokers (< 15 cigarettes/day), and 4.5 (95% CI = 2.3, 8.8) among heavier smokers. After adjustment for potential confounders, the relative risks were 1.4 (95% CI = 0.9, 2.4), 2.5 (95% CI = 0.9, 7.3), and 4.3 (95% CI = 2.2, 8.5), respectively. CONCLUSION: We found a positive, dose-related association between smoking and suicide among White men. Although inference about causality is not justified, our findings indicate that the smoking-suicide connection is not entirely due to the greater tendency among smokers to be unmarried, to be sedentary, to drink heavily, or to develop cancers. PMID- 10800429 TI - Trends in fruit and vegetable consumption among adults in 16 US states: Behavioral Risk Factor Surveillance System, 1990-1996. AB - OBJECTIVES: This study examined trends in fruit and vegetable consumption among adults in 16 US states. METHODS: Data from telephone surveys were used to stratify respondents by sociodemographic and health-related characteristics. RESULTS: The proportion of adults who consumed fruits and vegetables at least 5 times daily was 19%, 22%, and 23% in 1990, 1994, and 1996, respectively. While the proportion increased among those with active leisure-time physical activities and normal weight, it remained almost the same among inactive people and dropped among the obese. CONCLUSIONS: Progress in fruit and vegetable intake from 1990 to 1994 was encouraging, but it changed little between 1994 and 1996. PMID- 10800430 TI - An interactive CD-ROM for nutrition screening and counseling. AB - OBJECTIVES: The goal of this project was to develop an interactive CD-ROM for nutrition screening and counseling, designed to produce dietary behavior change in fat and fruit and vegetable intake. METHODS: The design was based on principles of relevance to the learner, readiness for change, feedback, individualization, facilitation of skills, and goal setting. It was tested in community settings such as libraries, senior centers, and Women, Infants, and Children clinics. RESULTS: Nearly 80% of the respondents (n = 284), including numerous low-income persons, reported learning something new about nutrition and health or their own dietary habits. More than 50% of those recontacted 2 to 4 weeks later had put some of their dietary goals into practice. CONCLUSIONS: This program is useful for dietary screening, feedback, skill building, and motivation in settings in which in-person counseling by nutrition professionals is not feasible. PMID- 10800431 TI - A brief smoking cessation intervention for women in low-income planned parenthood clinics. AB - OBJECTIVES: The purpose of this study was to evaluate a brief smoking cessation intervention for women 15 to 35 years of age attending Planned Parenthood clinics. METHODS: Female smokers (n = 1154) were randomly assigned either to advice only or to a brief intervention that involved a 9-minute video, 12 to 15 minutes of behavioral counseling, clinician advice to quit, and follow-up telephone calls. RESULTS: Seventy-six percent of those eligible participated. Results revealed a clear, short-term intervention effect at the 6-week follow-up (7-day self-reported abstinence: 10.2% vs 6.9% for advice only, P < .05) and a more ambiguous effect at 6 months (30-day biochemically validated abstinence: 6.4% vs 3.8%, NS). CONCLUSIONS: This brief, clinic-based intervention appears to be effective in reaching and enhancing cessation among female smokers, a traditionally underserved population. PMID- 10800432 TI - Cigars, youth, and the Internet link. AB - OBJECTIVES: This study examined the accessibility and appeal to youth of cigar marketing sites on the Internet. METHODS: Sites marketing cigars (n = 141) were examined for age restrictions, prices, health warnings, and other elements. RESULTS: Although it is illegal for minors to purchase tobacco, only 36 sites (25.5%) prohibited purchases by minors. Sites offered low prices, and 32% accepted money orders, cashier's checks, or cash-on-delivery (COD) orders. Almost 30% of the sites included elements with potential youth appeal; only 3.5% displayed health warnings. CONCLUSIONS: The unregulated promotion of cigars on the Internet has the potential to attract youth, and there are few barriers to Internet tobacco purchases by minors. PMID- 10800433 TI - The effect of the Taiwan motorcycle helmet use law on head injuries. AB - OBJECTIVES: This study evaluated the effect of the motorcycle helmet law implemented in Taiwan on June 1, 1997. METHODS: Collecting data on 8795 cases of motorcycle-related head injuries from 56 major Taiwanese hospitals, we compared the situation 1 year before and after implementation of the helmet law. RESULTS: After implementation of the law, the number of motorcycle-related head injuries decreased by 33%, from 5260 to 3535. Decreases in length of hospital stay and in severity of injury and better outcome were also seen. The likelihood ratio chi 2 test showed that severity decreased after the law's implementation (P < .001). Full helmets were found to be safer than half-shell helmets. CONCLUSION: The helmet law effectively decreased the mortality and morbidity from motorcycle related head injuries. PMID- 10800434 TI - Plastic wall materials in the home and respiratory health in young children. AB - OBJECTIVES: The relation between the presence of plastic wall materials in the home and respiratory health in children was assessed. METHODS: This population based cross-sectional study involved 2568 Finnish children aged 1 to 7 years. RESULTS: In logistic regression models, lower respiratory tract symptoms- persistent wheezing (adjusted odds ratio [OR] = 3.42, 95% confidence interval [CI] = 1.13, 10.36), cough (OR = 2.41, 95% CI = 1.04, 5.63), and phlegm (OR = 2.76, 95% CI = 1.03, 7.41)--were strongly related to the presence of plastic wall materials, whereas upper respiratory symptoms were not. The risk of asthma (OR = 1.52, 95% CI = 0.35, 6.71) and pneumonia (OR = 1.81, 95% CI = 0.62, 5.29) was also increased in children exposed to such materials. CONCLUSIONS: Emissions from plastic materials indoors may have adverse effects on the lower respiratory tracts of small children. PMID- 10800435 TI - Breast cancer screening in the United States and Canada, 1994: socioeconomic gradients persist. AB - OBJECTIVES: This study compared rates of annual mammography screening across socioeconomic status between the United States and Canada in 1994. METHODS: Population-based cross-sectional surveys were used to compare the rates. RESULTS: Screening rates were higher in the United States than in Canada for women aged 50 to 69 years (47.3% vs 38.8%; P < .01). Women with higher education and with higher incomes were more likely to receive screening in both countries, with no significant differences between countries. CONCLUSIONS: For women aged 50 to 69 years, screening rates in Canada have substantially increased relative to those in the United States. However, disparities in screening across levels of socioeconomic status persist in both countries. PMID- 10800436 TI - Recent data are needed to support public health training and workforce initiatives. PMID- 10800437 TI - The role of emergency obstetric care in the Safe Motherhood Initiative. PMID- 10800438 TI - The role of health services research in the Safe Motherhood Initiative. PMID- 10800439 TI - [Perspectives for surgery in the 21st century]. PMID- 10800440 TI - [Intravenous perfluorocarbons: 30 years of research--what now?]. PMID- 10800442 TI - [Laparoscopic splenectomy for idiopathic thrombocytopenia]. AB - BACKGROUND AND OBJECTIVE: Splenectomy is of great benefit for patients with idiopathic thrombocytopenia (ITP), when medical therapy has failed. It was the aim of this retrospective study to evaluate the results of laparoscopic splenectomy. PATIENTS AND METHODS: All patients with ITP, who had a laparoscopic splenectomy between 1992 and 1999, were included in this study. RESULTS: 53 patients with an average age of 41.8 years had a laparoscopic splenectomy: 48 of these operations were finished laparoscopically, while conversion to the open technique was necessary in five patients. The mean operation time was 95 minutes. In the last 38 cases it was 76 minutes. There were no deaths and no reoperations were necessary. The average postoperative hospital stay was 3.9 days. After a mean follow-up of 24 (1-75) months thrombocytopenia recurred in eight patients, of whom five received medical therapy. CONCLUSION: Splenectomy can be easily performed laparoscopically in patients with ITP and has a low morbidity. The results of therapy of ITP are good. Because of the quick convalescence the patients' comfort is better than with the conventional operation. PMID- 10800441 TI - [Cytomegalovirus infection after heart transplantation. Retrospective analysis of an antiviral CMV prevention]. AB - OBJECTIVE: Cytomegalovirus (CMV) infection is the most common viral infection in the early period after heart transplantation (HTX) and causes a significant morbidity and mortality. Although controversial, CMV is related to acute and chronic allograft rejection and to the development of graft vascular disease. It therefore plays an important role in the long-time outcome after solid organ transplantation. PATIENTS AND METHODS: 45 patients received a new heart between 1.1.97 and 31.12.1998. All of them were enrolled postoperatively in three-month antiviral prophylaxis (Cymevene). Only those patients were excluded from prophylaxis who were seronegative for CMV and received hearts from seronegative donors (n = 6). The pp65 antigenaemia assay and the murex hybrid capture CMV DNA assay on peripheral blood as well as the early antigen detection in the urine were used for CMV detection and also for monitoring. RESULTS: A total number of 580 assays were analysed (12.9 assays/patient). 561 tests (96.7%) were negative, 19 (3.3%) were positive. For CMV testing the pp65 antigenemia assay was used in 64.1%, the murex hybrid capture CMV DNA assay in 18.4% and the urine early antigen detection in 17.4%. Three patients (6.7%) developed viraemia during the first 3 postoperative months. Two patients (4.4%) suffered from CMV infection 8 and 9 months after heart transplantation and had to be treated with antiviral agents. Three patients (6.7%) died early after transplantation, but none had a CMV infection. CONCLUSION: Prevention of CMV disease was successful with three months of antiviral CMV prophylaxis after HTX. Asymptomatic viraemia during the prophylaxis period did not lead to tissue invasive disease. It is possible to carry out rapid CMV detection and CMV monitoring with the commercially available antigenaemia assays. PMID- 10800443 TI - [Thoracoscopic longitudinal myotomy of the esophagus. Successful alternative treatment for diffuse esophageal spasm]. AB - HISTORY AND ADMISSION FINDINGS: A 55-year-old woman had for 5 years been suffering from severe burning thoracic pain. She had typical risk factors for cardiovascular disease, namely hypertension, chronic smoking, obesity and hypercholesterolaemia. She had been hospitalized several times for suspected myocardial infarction, but coronary heart disease had been excluded by cardiac examination, including angiocardiography. The only contributory admission finding was epigastric pain on pressure. INVESTIGATION: Gastroscopy revealed a non erosive duodenitis and chronic antral gastritis. The cardia was tightly closed and there were no signs of reflux oesophagitis. Manometry in the oesophagus showed frequent simultaneous contractions and marked increase in pressure amplitude, predominantly in the distal segment, indicating diffuse oesophageal spasms. TREATMENT AND COURSE: Drug treatment with calcium channel blockers gave only brief relief from the thoracic pain. But longitudinal oesophageal myotomy via thoracoscopy brought about complete pain relief, which has now persisted for 3 years. CONCLUSION: Thoracoscopic longitudinal oesophageal myotomy can provide a therapeutic alternative in patients with oesophageal spasms when drug therapy and pneumatic oesophageal dilatation have failed. PMID- 10800444 TI - [Neuroendocrine tumors of the gastroenteropancreatic system. Diagnosis and surgical treatment]. PMID- 10800446 TI - [Extracorporeal shockwave lithotripsy for cholecystolithiasis using a new mini lithotripter]. PMID- 10800445 TI - [Intravenous perfluorocarbons. Artificial oxygen carriers and their medical applications]. PMID- 10800447 TI - Professor Edwin Haslam, recipient of the 3rd Tannin Conference Award. PMID- 10800448 TI - Che faro senza polifenoli? PMID- 10800449 TI - Highly oxidized ellagitannins and their biological activity. PMID- 10800450 TI - NMR characterization and biological evaluation of proanthocyanidins: a systematic approach. PMID- 10800451 TI - Chemistry, biosynthesis, and biological activity of natural Diels-Alder type adducts from moraceous plants. PMID- 10800452 TI - Birdsfoot trefoil: a model for studying the synthesis of condensed tannins. PMID- 10800453 TI - Isotopic labelling of dietary polyphenols for bioavailability studies. PMID- 10800454 TI - Human metabolism of dietary quercetin glycosides. PMID- 10800455 TI - Spin stabilizing approach to radical characterization of phenylpropanoid antioxidants: an ESR study of chlorogenic acid oxidation in the horseradish peroxidase, tyrosinase, and ferrylmyoglobin protein radical systems. PMID- 10800456 TI - Polyphenols increase adhesion between lipid bilayers by forming interbilayer bridges. PMID- 10800458 TI - Glycosylation, esterification and polymerization of flavonoids and hydroxycinnamates: effects on antioxidant properties. PMID- 10800457 TI - Antiatherogenic effects of tea polyphenols (flavan-3-ols) in humans and apoE deficient mice. PMID- 10800459 TI - Tannins as biological antioxidants. PMID- 10800460 TI - Interaction of flavanoids with peptides and proteins and conformations of dimeric flavanoids in solution. PMID- 10800461 TI - Modeling the conformation of polyphenols and their complexation with polypeptides: self-association of catechin and its complexation with L-proline glycine oligomers. PMID- 10800462 TI - Polyphenols, metal ion complexation and biological consequences. PMID- 10800463 TI - Anti-caries activity of bark proanthocyanidins. PMID- 10800464 TI - Enhancement of antimicrobial activity of tannins and related compounds by immune modulatory effects. PMID- 10800465 TI - Induction of apoptosis and anti-HIV activity by tannin- and lignin-related substances. PMID- 10800466 TI - Antitumor activities of ellagitannins on tumor cell lines. PMID- 10800467 TI - Chemical constituents of mainly active component fractionated from the aqueous tea non-dialysates, an antitumor promoter. PMID- 10800468 TI - Host-mediated anticancer activities of tannins. PMID- 10800469 TI - Inhibitory effects of hydrolyzable tannins on tumor promoting activities induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in JB6 mouse epidermal cells. PMID- 10800470 TI - Plant lignans and health: cancer chemoprevention and biotechnological opportunities. PMID- 10800471 TI - Functional properties of hop polyphenols. PMID- 10800472 TI - Modification of the solubility of tannins: biological significance and synthesis of lipid-soluble polyphenols. PMID- 10800473 TI - Phenolic content and antioxidant activity: a study on plants eaten by a group of howler monkeys (Alouatta fusca). PMID- 10800474 TI - Tannins as nutritional constraints for elk and deer of the coastal Pacific Northwest. PMID- 10800475 TI - Antimetabolites. PMID- 10800476 TI - Bleomycin. PMID- 10800477 TI - Mitomycins. PMID- 10800478 TI - Taxanes and other microtubule stabilizing agents. PMID- 10800479 TI - DNA topoisomerase I poisons. PMID- 10800480 TI - DNA topoisomerase II poisons and inhibitors. PMID- 10800481 TI - Cisplatin. PMID- 10800482 TI - Multidrug resistance. PMID- 10800483 TI - New anticancer agents. PMID- 10800484 TI - Monoclonal antibodies, cytokines and fusion proteins in the treatment of malignant disease. PMID- 10800485 TI - Biological response modifiers. PMID- 10800486 TI - Adoptive immunotherapy. PMID- 10800487 TI - Hematopoietic growth factors in cancer chemotherapy. AB - Hematopoietic growth factors have made a significant impact on the treatment of cancer, primarily in the prevention of infections associated with chemotherapy induced neutropenia, in progenitor cell transplantation, in chemotherapy-induced thrombocytopenia, and in chemotherapy-induced anemia. As seen with this review, our basic understanding of hematopoietic growth factors and their clinical potential continue to expand. Work will need to continue, especially with the new thrombopoietic factors, megakaryocyte growth and developing factor, thrombopoietin, and IL-11, to fully categorize their biology and clinical characteristics. PMID- 10800488 TI - Leukemias and plasma cell myeloma. PMID- 10800489 TI - Non-Hodgkin's lymphomas. PMID- 10800490 TI - Head and neck cancer. PMID- 10800491 TI - Lung cancer. AB - The results of the many clinical trials published in 1997 had only modest impact on the treatment results using either cytostatic agents alone or combined with radiotherapy in lung cancer. In SCLC, combination chemotherapy including platin compounds (cisplatin, carboplatin) and the podophyllotoxins (etoposide and teniposide) continue to be the cornerstone of therapy. Complete plus partial responses to combination chemotherapy occurs in 80-90% of all patients. Median survival is at present 11-17 months, and overall 5-year survival approximately 5% depending on the initial tumor stage. The duration of treatment has been shortened to 5-6 months. In relapsing patients topotecan and paclitaxel appear to have clinical useful activity. For epidermoid, adeno- and large cell carcinoma further support has arisen for the use of preoperative and preirradiatory chemotherapy in stage III NSCLC, but further studies are needed before specific recommendations can be given for general use. For patients with advanced NSCLC, new innovative treatments are still urgently needed. Platin-containing regimens with the inclusion of new agents, such as gemcitabine, taxenes, and navelbine, yielded response rates of 42-55% in phase II trials, but results from large phase III trials are necessary in order to measure the impact of these new agents in the management of NSCLC. Major improvements of therapy for mesothelioma have not occurred within the last year. PMID- 10800492 TI - Upper gastrointestinal tumors. PMID- 10800493 TI - Cancers of the large bowel and hepatobiliary tract. PMID- 10800494 TI - Endocrine tumors. PMID- 10800495 TI - Genitourinary malignancies. PMID- 10800496 TI - Gynecological cancers. PMID- 10800497 TI - Breast cancer. PMID- 10800498 TI - Melanoma. AB - In melanoma, conventional therapies, especially cytotoxic chemotherapies, have proven unsatisfactory. Although a variety of agents have been tested singly and in combination, recent randomized studies have demonstrated that the response rates observed in single institution phase II trials are not generalizable to multi-institution settings. Studies of cytokine therapy, especially interferon alpha in the adjuvant situation, and interleukin-2 for advanced disease, have demonstrated some utility to these agents. However, their toxicities remain formidable, and studies refining their use are under way. The limited successes of such immunomodulatory strategies have provided a rationale for continued pursuit of immunotherapy approaches. It is likely that in the near future such active immunization protocols will continue to be actively investigated. PMID- 10800499 TI - Soft tissue and bone sarcomas. PMID- 10800500 TI - Brain tumors. PMID- 10800501 TI - Biology and therapy of pediatric malignant solid tumors. PMID- 10800502 TI - Supportive care. PMID- 10800503 TI - The future's bright for 2000 and beyond: swine veterinarians, industry making strong comeback. PMID- 10800504 TI - National standards for organic foods proposed. PMID- 10800505 TI - Debating vivisection within the curriculum. PMID- 10800506 TI - Debating vivisection within the curriculum. PMID- 10800507 TI - What is your diagnosis? Fracture of the medial extremity of the distal sesamoid (navicular) bone. PMID- 10800508 TI - When is there a duty to treat animals? PMID- 10800509 TI - Oral vesicular lesions in horses without evidence of vesicular stomatitis virus infection. AB - OBJECTIVE: To report clinical and serologic findings in horses with oral vesicular lesions that were consistent with vesicular stomatitis (VS) but apparently were not associated with VS virus (VSV) infection. DESIGN: Serial case study. ANIMALS: 8 horses. PROCEDURE: Horses were quarantined after appearance of oral lesions typical of VS. Severity of clinical signs was scored every 2 to 5 days for 3 months. Serum samples were tested for antibodies by use of competitive ELISA (cELISA), capture ELISA for IgM, serum neutralization, and complement fixation (CF). Virus isolation was attempted from swab specimens of active lesions. RESULTS: 2 horses with oral vesicular lesions on day 1 had antibodies (cELISA and CF) against VSV; however, results of CF were negative by day 19. Five of the 6 remaining horses were seronegative but developed oral lesions by day 23. Virus isolation was unsuccessful for all horses. CONCLUSIONS AND CLINICAL RELEVANCE: Horses were quarantined for 75 days in compliance with state and federal regulations. However, evidence suggests that oral lesions were apparently not associated with VSV infection. The occurrence in livestock of a vesicular disease that is not caused by VSV could confound efforts to improve control of VS in the United States and could impact foreign trade. Vesicular stomatitis is of substantial economic and regulatory concern. PMID- 10800510 TI - Sexual harassment in the veterinary academic environment. AB - OBJECTIVE: To survey faculty and house officers of clinical departments of colleges of veterinary medicine (CVM) to identify characteristics of sexual harassment (SH) in the veterinary academic environment, to report the opinions of survey respondents on how SH is being handled, and to determine how the process can be improved at veterinary academic institutions. PROCEDURE: On the basis of lists obtained from 25 CVM, a survey was mailed to 1,294 academic veterinarians. Four hundred seventy-eight completed surveys were returned. RESULTS: The prevalence of SH in the population of respondents was 31%. Nonphysical forms of SH were reported 6 times as often as physical forms of harassment, with the most common type reported being offensive sexual comments and unwanted attention. Fear of reprisal was the most prevalent reason cited by respondents for not confronting the harasser. Survey respondents rated the following as very important to improve the system of dealing with SH at their academic institution: guarantee of protection from retaliation, assurance of confidentiality, clear explanation of what will happen to you, and a clearer definition of SH. CONCLUSION: A clear definition of SH is the first step in preventing SH. Other cited steps include professional development programs to educate the academic population as to what constitutes SH, inform the entire academic population what the institution's SH policy is, and enforce this policy with sensitivity, fairness, confidentiality, and quick resolve to protect the victim. PMID- 10800511 TI - Breed distribution of dogs with diabetes mellitus admitted to a tertiary care facility. AB - OBJECTIVE: To determine which dog breeds are at low and high risk for developing diabetes mellitus (DM). DESIGN: Cohort study. ANIMALS: Hospital population of 221 dogs with DM and 42,882 dogs without DM during 5.5 years. PROCEDURE: 165 breeds (including a mixed-breed category) were represented in the hospital population. Breed-specific expected numbers of dogs with DM were calculated by multiplying the proportion of all dogs admitted to the hospital that were determined to have DM during the study period by the breed-specific totals during the study period. Breeds or breed groups evaluated in the analysis (n = 20) were restricted to those that had a combined observed and expected count > 5 to document breeds at low and high risk for developing DM. Proportionate changes in the risk of developing DM by breed were calculated and presented using exact odds ratios, 95% confidence intervals, and P values. Mixed-breed dogs were chosen as the reference breed. RESULTS: Samoyeds, Miniature Schnauzers, Miniature Poodles, Pugs, and Toy Poodles were at high risk for developing DM. Dog breeds found to be at low risk for developing DM were German Shepherd Dog, Golden Retriever, and American Pit Bull Terrier. CONCLUSION AND CLINICAL RELEVANCE: The finding that certain dog breeds are at low or high risk for developing DM suggests that some genetic defects may predispose dogs to development of DM, whereas other genetic factors may protect dogs from development of DM. PMID- 10800512 TI - Evaluation of a commercial enzyme-linked immunosorbent assay for detection of Borrelia burgdorferi exposure in dogs. AB - OBJECTIVE: To evaluate the effectiveness of a commercially available ELISA kit for detecting antibodies against Borrelia burgdorferi in dogs. SAMPLE POPULATION: Banked sera from 440 military working dogs were used for serologic analyses. PROCEDURE: Serum samples were analyzed for antibodies against B burgdorferi by use of a commercially available ELISA and subsequently by western blot analysis as a confirmatory test. RESULTS: Results from the ELISA indicated that 89 (20%) samples were positive for exposure to B burgdorferi or canine Lyme disease vaccine, and 351 (80%) were negative. Follow-up testing by western blot analysis indicated that results for 109 (25%) samples were positive and 331 (75%) were negative for exposure. All samples that had positive results on the ELISA also had positive results on western blot analysis (true positives). Of the 351 samples that had negative results on the ELISA, only 331 had negative results on western blot analysis (true negatives). The remaining 20 samples had positive results on western blot analysis. By use of a standard 2 x 2 table, it was determined that the ELISA had a sensitivity of 82%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 94%. CONCLUSIONS AND CLINICAL RELEVANCE: The commercial ELISA kit evaluated in this study appeared to lack adequate sensitivity for detecting all potential cases of borreliosis in dogs. The ELISA was also unable to discriminate natural exposure from exposure attributable to vaccination, which could complicate interpretation of positive results and treatment of dogs with clinical signs. PMID- 10800513 TI - Severe hypercalcemia in a dog with a retained fetus and endometritis. AB - A 10-month-old sexually intact female German Shorthaired Pointer examined because of lethargy, episodes of fever, inappetence, and vomiting was found to have severe hypercalcemia. Results of laboratory testing, radiography, and ultrasonography excluded previously recognized causes of hypercalcemia in dogs. Instead, the dog was found to have purulent endometritis and an incompletely resorbed fetus. Treatment with fluids i.v., diuretics, and calcitonin failed to adequately reduce serum calcium concentration, but serum calcium concentration was normal within 4 days after the dog underwent an ovariohysterectomy. Retention of one or more fetuses and endometritis should be included in the differential diagnosis for dogs with hypercalcemia. PMID- 10800514 TI - Acute ibuprofen toxicosis in a ferret. AB - A 20-month-old 1.44-kg (3.2-lb) castrated male ferret was examined because of vomiting, defecating in an abnormal location, dyspnea, and signs of depression. Within 5 minutes of initial evaluation, the ferret became nonresponsive to all stimuli except deep pain. Despite intensive supportive treatment, the ferret died. Toxicologic analyses for ibuprofen were performed on serum, urine, and liver, using gas chromatography and mass spectrophotometry. Serum ibuprofen concentrations were 245 micrograms/g before and 269 micrograms/g after death. Acute ibuprofen toxicosis may cause severe lethargy progressing to coma, apnea, and death in ferrets. Ibuprofen toxicosis should be considered in differential diagnoses for ferrets with signs of depression, with or without clinical signs of gastrointestinal tract dysfunction. PMID- 10800515 TI - Idiopathic pure red cell aplasia and nonregenerative immune-mediated anemia in dogs: 43 cases (1988-1999). AB - OBJECTIVE: To examine clinical features, laboratory test results, treatment, and outcome of dogs with pure red cell aplasia (PRCA) and idiopathic nonregenerative immune-mediated anemia (NRIMA). DESIGN: Retrospective study. ANIMALS: 43 dogs with severe nonregenerative anemia. PROCEDURE: Medical records of dogs determined to have PRCA, NRIMA, or ineffective erythropoiesis on the basis of bone marrow analysis between 1988 and 1999 were reviewed. Criteria for inclusion were > or = 5-day history of severe nonregenerative anemia (Hct < 20%; < 60.0 x 10(3) reticulocytes/microliter) with no underlying diseases. Information was retrieved on signalment, clinical signs, laboratory test results, treatment, and outcome. RESULTS: Median age of the dogs was 6.5 years. Spayed females and Labrador Retrievers were significantly overrepresented. Median Hct was 11% with no evidence of regeneration (median, 1.5 x 10(3) reticulocytes/microliter). Direct Coombs' test results were positive in 57% of dogs. Biochemical abnormalities included hyperferremia and high percentage saturation of transferrin. Bone marrow findings ranged from PRCA (5%) to erythroid hyperplasia (55%). Myelofibrosis was common. Dogs were treated with immunosuppressive drugs and the response was complete, partial, and poor in 55, 18, and 27% of the dogs, respectively. Mortality rate was 28%. CONCLUSIONS AND CLINICAL RELEVANCE: An immune-mediated pathogenesis should be considered in dogs with severe, nonregenerative anemia, normal WBC and platelet counts, hyperferremia, mild clinical signs, and no evidence of underlying disease. Bone marrow findings range from the rare PRCA to erythroid hyperplasia. Myelofibrosis is often detected in affected dogs and may prevent bone marrow aspiration. PMID- 10800516 TI - Endogenous lipid pneumonia in cats: 24 cases (1985-1998). AB - OBJECTIVE: To determine clinical signs, radiographic and histologic abnormalities, and concurrent diseases in cats with endogenous lipid pneumonia (EnLP) and to determine the pathologic importance of EnLP in cats. DESIGN: Retrospective study. ANIMALS: 24 cats. PROCEDURE: Medical records of cats in which EnLP was confirmed by histologic examination of necropsy specimens were reviewed. Information collected from the medical records included signalment, body weight, clinical signs, and results of clinicopathologic tests. Thoracic radiographs were reviewed by a radiologist; histologic specimens were reviewed by a pathologist. RESULTS: All cats had nonspecific clinical abnormalities, such as lethargy, anorexia, or weight loss; 16 had signs of respiratory tract disease. All cats had concurrent systemic diseases, and clinicopathologic abnormalities were reflective of these conditions. Nonspecific abnormalities were detected on thoracic radiographs from 9 of 11 cats. Most cats had macroscopic, multifocal, subpleural lesions; inflammatory infiltrates, cholesterol clefts, and multinucleated giant cells were common. Ten cats had an underlying obstructive pulmonary disease that was the likely cause of EnLP. Lesions of EnLP were not considered to be severe enough or extensive enough to be the cause of death in any of these cats. CONCLUSIONS AND CLINICAL RELEVANCE: EnLP is an uncommon respiratory tract disorder of cats with no pathognomonic clinical, laboratory, or radiographic findings. Although EnLP was not the cause of death in any of these cats, results of the present study do suggest that EnLP may be a marker for potentially severe underlying obstructive pulmonary disease. PMID- 10800517 TI - Prevalence and clinical importance of heart murmurs in racehorses. AB - OBJECTIVE: To determine the prevalence of various types of heart murmurs in Thoroughbred racehorses and assess their association with performance by echocardiography and review of the horses' race records for the preceding 2 years. DESIGN: Clinical and retrospective study. ANIMALS: 846 Thoroughbred racehorses. PROCEDURE: Cardiac auscultations were performed by 3 individuals; for 30 horses, Doppler echocardiographic examinations were also performed. Statistical analyses of race records for 753 horses were performed to assess association of heart murmurs with performance. RESULTS: Heart murmurs were detected by cardiac auscultation in 686 of 846 (81.1%) horses. Systolic murmurs over the heart base were most common; 365 (43.1%) horses had systolic murmurs that were loudest over the pulmonary valve area, and 232 (27.4%) horses had systolic murmurs that were loudest over the aortic valve area. Systolic murmurs over the tricuspid valve area were detected in 241 (28.5%) horses, whereas systolic murmurs over the mitral valve area were detected in only 32 (3.8%) horses. Diastolic murmurs were much less common than systolic murmurs. Review of race records did not reveal a significant association between murmurs and performance. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that heart murmurs are a common finding in racehorses; most of these heart murmurs do not appear to be clinically important. PMID- 10800518 TI - Mesenteric rents as a source of small intestinal strangulation in horses: 15 cases (1990-1997). AB - OBJECTIVE: To describe the clinical findings in horses with small intestinal strangulation through mesenteric rents, and to determine the recurrence and survival rates after surgery. DESIGN: Retrospective study. ANIMALS: 15 horses with small intestinal obstruction via a mesenteric rent. PROCEDURE: Medical records of horses with obstruction of the small intestine via a mesenteric rent between January 1990 and December 1997 were reviewed. The signalment, history, initial physical examination findings, results of abdominocentesis, and clinical laboratory values were recorded. Surgical findings, including location of the mesenteric rent and surgical procedure performed, were recorded. Short- and long term survival rates were calculated. RESULTS: Most mesenteric rents were located in the mesentery of the small intestine (13 horses). Two horses had multiple mesenteric defects. Seven horses were euthanatized at surgery because of an inability to reduce the entrapped intestine (3 horses), uncontrollable hemorrhage (2), inability to close the rent (1), and the amount of compromised intestine involved (1). Seven horses required intestinal resection and anastomosis. The median length of intestine resected was 2.6 m (range, 0.6 to 4.5 m). The mesenteric rents created during resection were not closed in 2 horses. One of these 2 horses subsequently developed a strangulating obstruction through the open rent. Seven of 15 horses in our study were discharged from the hospital (i.e., short-term survival rate of 47% [7/15]). Long-term follow-up information was available for 5 of the 7 horses (follow-up duration of 5 months to 9 years), of which 2 died as a result of colic, and 1 horse was euthanatized because of severe arthritis (i.e., long-term survival rate of 40% [2/5]). CONCLUSIONS AND CLINICAL RELEVANCE: Inability to reduce the intestinal obstruction, severe hemorrhage from the mesentery, and the length of intestine involved are the main factors that decrease survival rates in horses with small intestinal strangulation caused by mesenteric rents. PMID- 10800519 TI - Herd-level risk factors for infection with Mycobacterium paratuberculosis in US dairies and association between familiarity of the herd manager with the disease or prior diagnosis of the disease in that herd and use of preventive measures. AB - OBJECTIVE: To evaluate associations among herd infection status, herd management practices, and familiarity of the herd manager with Mycobacterium paratuberculosis (Johne's disease) or prior disease diagnosis in that herd to support development of Johne's disease-control programs. DESIGN: Population-based cross-sectional study. SAMPLE POPULATION: 1,004 US dairies, each with > or = 30 cows, representing 79.4% of US dairy cows. PROCEDURE: Questionnaires were administered to dairy managers, and blood samples were collected from cows during herd visits. Sera were tested for antibodies to M paratuberculosis, using a commercially available ELISA. Multivariable logistic regression analyses were used to evaluate associations between use of management practices, herd disease status, and familiarity of the manager with Johne's disease or prior diagnosis of Johne's disease in that herd. RESULTS: Results from serologic testing revealed that 3.4% of cows and 21.6% of dairy herds were infected with M paratuberculosis. Factors associated with infection included number of cows in herd, region of country, percentage of cows born at other dairies, group housing for periparturient cows, and group housing for preweaned calves. Few preventive practices were positively associated with prior diagnosis of Johne's disease (time of separation of newborn calf from dam) or familiarity of the manager with the disease (teats and udder washed before colostrum collected). CONCLUSIONS AND CLINICAL RELEVANCE: Risk factors associated with Johne's disease in this study confirmed those management practices generally recommended for disease control. An educational problem, however, is the finding that herd managers familiar with Johne's disease generally use management practices similar to those used by managers unfamiliar with the disease. PMID- 10800520 TI - Nucleoside triphosphate-binding proteins: different scaffolds to achieve phosphoryl transfer. PMID- 10800521 TI - NMR studies of protein-nucleic acid complexes: structures, solvation, dynamics and coupled protein folding. PMID- 10800522 TI - Therapeutic implications of recent data on immune pathogenesis of HIV disease. PMID- 10800523 TI - Surviving the fall: an AIDS caregiver's journey. PMID- 10800524 TI - Kala-azar as an AIDS-related opportunistic infection. AB - Visceral leishmaniasis (kala-azar) is a worldwide disseminated protozoal infection primarily transmitted by sand flies. Because host defense against this intracellular infection is T-cell-dependent, kala-azar has predictably joined the list of AIDS-related opportunistic infections in endemic areas. The vast majority of patients with AIDS-associated kala-azar are currently found in southern Europe (the Mediterranean basin, especially Spain in injection drug users); future cases will inevitably arise in other endemic regions including India, East Africa and Sudan, and Brazil. In CD4 cell-deficient HIV-infected individuals, kala-azar likely represents recrudescence of previously controlled asymptomatic infection; in drug users, newly acquired infection may result from transmission via shared needles. Coinfected patients are frequently parasitemic and may show atypical clinical presentations, unusual multi-organ involvement, and absent antileishmanial antibodies. Diagnosis is made by microscopic examination or culture of aspirate or biopsy of any involved tissue (primarily bone marrow) or by blood smear or culture. Conventional treatment (pentavalent antimonials) induces initial remission in about 50% of patients; amphotericin B and its new lipid formulations appear more active. If suppressive maintenance therapy is not used, relapse within 1 year is typical. In AIDS patients with a first episode of visceral kala-azar, up to 25% die within 1 month if treatment is stopped. Optimal primary and secondary prophylaxis for AIDS-related kala-azar remain to be determined; life-long maintenance therapy is becoming an accepted approach. PMID- 10800525 TI - Rifabutin prophylaxis against Mycobacterium avium complex infections in HIV infected patients: impact on the incidence of campylobacteriosis. AB - Following the observation of the decreasing occurrence of campylobacteriosis in HIV-infected patients. This study examines the incidence of campylobacteriosis in patients who had received rifabutin prophylaxis against Mycobacterium avium complex (MAC) infection compared with the incidence observed among patients treated before the advent of rifabutin. A retrospective analysis (February 1992 to November 1995) was conducted in a hospital HIV inpatient unit. The study included two patient groups: 73 HIV-infected patients with CD4 counts of < 100 cells/microL (mean 30 cells/microL) who were treated between February 1992 and July 1993 and who had not received rifabutin prophylaxis (Group R-), as well as 90 HIV-infected patients with CD4 counts of < 100 cells/microL (mean 22 cells/microL) who had received rifabutin 300 mg/day as primary prophylaxis against MAC bacteremia between July 1993 and November 1995 (Group R+). For the patient population as a whole, 20 episodes of campylobacter infection were observed in 13 patients. Causative pathogens were Campylobacter jejuni (n = 10), C. coli (8), and unidentifiable (2). Seventeen episodes (in 12 patients) of campylobacter infection occurred in Group R- versus 3 episodes (in 2 patients) in Group R+ (p < 0.0005). The rate of symptomatic infection per 100 patient-months was 0.251 in Group R+ versus 2.02 in Group R-. The results of this study indicate that rifabutin prophylaxis was associated with a decrease in the rate of campylobacter infection in HIV-infected patients. These findings are supported by evidence that rifabutin is active against C. jejuni in vitro. PMID- 10800526 TI - Clinician judgement as a tool for targeting HIV counseling and testing in North Carolina state mental hospitals, 1994. AB - HIV infection increasingly affects populations that may not appear at high risk based on the use of some traditional targeting strategies. To shed some light on how to more sensitively/effectively identify people who need routine HIV testing and counseling, the objective of this study is to determine the prevalence of HIV infection in North Carolina state mental hospitals and to evaluate clinician judgment as a tool for targeting HIV counseling and testing. The design used is a blinded seroprevalence study. The study population includes all patients admitted to North Carolina state mental hospitals between March 1st and May 31st, 1994. The main outcome measures are the HIV seroprevalence, demographic and diagnostic features, and clinician assessment of the likelihood of HIV infection. The results of the study find that of 2159 study subjects, 35 persons (1.6%) were infected with HIV; of these, 14 (40%) were not previously known to be infected. All 35 HIV infections occurred in persons aged 13-59 years. Within this age group, infection rates were significantly higher for Blacks, males, persons who had a diagnosis of organic brain disease, and persons who had multiple psychiatric diagnoses. However, testing strategies that targeted any of the higher risk groups were insensitive. The rate of HIV infection for persons judged by the admitting clinician to have a high or intermediate likelihood of HIV infection was 26.4 times higher than the rate for those judged to have a low likelihood of infection (2.1 vs. 0.1%, 95% confidence intervals: 3.5-201.3). Of the 14 previously undiagnosed HIV-infected persons, 13 were judged by clinicians to have a high or intermediate likelihood of HIV infection. Moreover, 1258 persons were correctly assessed to have a low likelihood of infection. Conclusions from this study are that an HIV counseling and testing strategy targeting persons (in this setting aged 13-59 years) who were judged by clinicians to have a high or intermediate likelihood of infection, would have identified more than 90% of previously undetected infections while substantially reducing the number of negative HIV tests performed. PMID- 10800527 TI - Home away from home: essential elements in developing housing options for people living with HIV/AIDS. AB - There is a paucity of data to enlighten our understanding regarding the need for a respite care/supported living home for people living with HIV/AIDS (PLWHIV/AIDS) and to identify the essential elements required to initiate this endeavor. Participatory action research was used in this study as the process by which the study was conducted. Focus group and in-depth interviews with 78 individuals were completed and subjected to qualitative thematic analysis. The findings suggest that a paucity of choices exist for PLWHIV/AIDS when they can no longer live independently in their own homes. Principles for developing HIV/AIDS housing programs are recommended, highlighting the need for active involvement of PLWHIV/AIDS in the program planning process. It is only with this collaboration and participation that the housing and healthcare needs of PLWHIV/AIDS can be adequately addressed. PMID- 10800528 TI - Use of stage of change (SOC) to develop an STD/HIV behavioral intervention: phase 1. A system to classify SOC for STD/HIV sexual risk behaviors--development and reliability in an STD clinic. AB - This paper reports on the initial phase in the development, program implementation, and inter-rater reliability of an application of Stage of Change (SOC) behavioral theory for use in STD/HIV risk reduction. SOC was adapted to assess readiness for sexual behavior change in an urban STD clinic in Rochester, New York. A standardized staging grid and protocol were developed and implemented as part of HIV pre- and post-test counseling. A "client instructor" methodology was used to assess standardization and staff inter-rater reliability. Percent agreement for stage assessment for target behaviors was calculated. The Rochester STD/HIV Behavioral Counseling (RoSHBeC) Staging Grid and Protocol was used to train staff and this staging system was implemented in the STD clinic. After training, staff identified the correct behavioral target and stage 90% of the time. Inter-rater agreement for SOC classification was greater than 70%. Our experience demonstrates that it is possible to develop, implement, and sustain an integrated provider-delivered STD/HIV behavioral intervention in a busy urban STD Clinic. This staging system has the potential for use in other settings and for other health-related behaviors. PMID- 10800529 TI - College educated most likely to get PIs. PMID- 10800530 TI - Efavirenz approved in Europe. PMID- 10800531 TI - AZT incorporates into leukocyte DNA. PMID- 10800533 TI - Treatment of vaginal infections. PMID- 10800532 TI - Nevirapine and methadone withdrawal. PMID- 10800534 TI - Disease may be sexually transmitted. PMID- 10800535 TI - Nose spray for gonorrhea. PMID- 10800536 TI - Genital warts vaccine. PMID- 10800538 TI - Sex education in schools supported. PMID- 10800537 TI - Protein effective against KS. PMID- 10800539 TI - Stress and lack of social support hastens HIV disease: most compelling evidence to date. PMID- 10800544 TI - [Enterovesical fistula in diverticulitis]. AB - Three men aged 52, 60, and 38 years, complained of dysuria and recurrent cystitis. One patient mentioned pneumaturia, a feature which later appeared to be present in all three. Ultrasound examination showed air in the bladder, and demonstrated the fistulous tract in two cases. Computed tomography identified the third fistula. In all 3 the enterovesical fistula was a complication of sigmoid diverticulitis. After sigmoid resection they recovered well. The key to the diagnosis of enterovesical fistula is to think of it. Pneumaturia and faecaluria are pathognomonic symptoms. Both ultrasound and computed tomography may be helpful in the diagnosis. PMID- 10800545 TI - [Identification of the gene for hyper-IgD syndrome: a model of modern genetics]. AB - Hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS) is a rare autosomal recessive disorder. Patients suffer from recurrent attacks (3-6 days) with fever, abdominal distress, lymphadenopathy, skin lesions and arthralgias. Patients display a constantly elevated serum IgD which serves as a biological marker of the disease. Recently, the gene for HIDS was discovered by two independent groups using positional and functional cloning methods. One group used linkage analysis (positional cloning) and was able to locate the gene for HIDS on the long arm of chromosome 12 (12q24). Mevalonate kinase was an interesting candidate gene because patients with a near complete absence of this enzyme (mevalonic aciduria) do exhibit attacks of fever. Indeed subsequent data showed that there was a decreased enzyme activity due to missense mutations in the mevalonate kinase gene. The other group detected slightly elevated urinary excretion of mevalonic acid during attacks in a HIDS patient (functional cloning). The enzyme activity of mevalonate kinase was lower in cultured cells and sequence analysis identified several missense mutations in cDNA encoding for mevalonate kinase. Mevalonate kinase is a key enzyme in the cholesterol synthesis pathway and it is rather surprising that a defect in the cholesterol metabolism can cause a periodic inflammatory disease such as HIDS. PMID- 10800546 TI - [Dendritic cells as carriers of human immunodeficiency virus (HIV)]. AB - In two papers in Cell researchers of the University Medical Center St. Radboud in Nijmegen (in collaboration with Utrecht and US researchers) have published the results of studies dealing with dendritic cells and their role in the pathogenesis of HIV-1 infection. First, they identified a dendritic cell receptor, DC-SIGN, that mediates adhesion with T cells. Second, they described another property of DC-SIGN, i.e. binding to the HIV-1 envelope protein gp120. Through this binding dendritic cells can transport HIV-1 to the CD4+ cells in lymphoid organs. Interestingly, it appeared that dendritic cells themselves are not infected by HIV-1, but only act as carriers for HIV-1. These findings may have important consequences for the design of new options for intervention in HIV 1 infection. PMID- 10800547 TI - [Fibroids and fertility]. AB - Fibroids are the most common benign uterine tumours. A relation between fibroids and infertility is suggested. Successful implantation depends on a normal, receptive endometrium. Fibroids can change the normal appearance of the uterine cavity and can adversely affect the endometrium. It seems that treatment of fibroids in infertility patients is useful but this is not yet proven scientifically. It is advised to refrain from extirpation of fibroids in patients with fertility problems. PMID- 10800548 TI - [Gastrointestinal surgery and gastroenterology. VIII. Gastroenterologic aspects of chronic gastrointestinal ischemia]. AB - The main cause of chronic gastrointestinal ischaemia is atherosclerosis. Stenotic lesions of the mesenteric circulation are relatively common, but lead to chronic ischaemic complaints due to collateral circulation in probably only 2-3 per 100,000 inhabitants per year. The classical presentation (post-prandial abdominal pain, weight loss, upper abdominal souffle) is present in a minority of patients only. Symptoms also occur after exercise. Gastric ulcers and diarrhoea are less frequent. Although patients with 2 and 3 vessel involvement (coeliac artery, superior mesenteric artery and inferior mesenteric artery) usually experience the most severe ischemic complaints, patients with single vessel involvement can also develop symptoms. In the diagnosis of cases with abdominal complaints, factors that aggravate or reduce the complaints anamnestically are the guideline for supplementary diagnostics. The more frequent causes of the symptoms are to be excluded first. Doppler-ultrasonography of the mesenteric vessels can detect most stenotic lesions accurately. To establish the diagnosis visceral angiography is needed. A new method of examination is magnetic resonance angiography (MRA). Another new method is tonometry during exercise: a PCO2 value in the lumen that is higher than that in the blood indicates ischaemia. Non-invasive treatment of chronic gastrointestinal ischaemia is aimed at reduction of the gastrointestinal metabolic workload by smaller meals, at suppression of acid secretion, at inhibition of the secretion of gastric acid and on risk factors for atherosclerosis. PMID- 10800549 TI - [Roaming through methodology. XVIII. Selection of control groups]. AB - The selection of a valid control group is important in case control studies and has therefore generated a fair amount of discussion in the epidemiologic literature. The proposal of the study usually determines the type of control group that should be selected. It is possible to select population controls, hospital controls, friends or relatives of patients or a variant of these. Each method has specific advantages and disadvantages. Random-digit dialing can be used to select population controls by telephone. This method is a valuable alternative when no registry exists. PMID- 10800550 TI - [Custom-made gold plate prosthesis in the upper eyelid in patients with facial paralysis: improved eyelid closure with less effects on other eye complaints]. AB - OBJECTIVE: To obtain insight into the results of treatment of lagophthalmos by gold weight implantation in the upper eyelid. DESIGN: Retrospective. METHODS: The records of all 37 patients treated in the Netherlands (1986-1998) were reviewed. All traceable patients (n = 33) were sent a questionnaire and invited for clinical evaluation of the treated eye (n = 28). RESULTS: The average follow-up was 44 months (range: 3-150). Twenty-seven patients (82%) reported improved eye closure. After 6 weeks 13 patients (48%) had no eye complaints (dry eye, epiphora, conjunctivitis). At the clinical evaluation 10 patients (36%) had no such complaints. Nine patients (27%) had to undergo a reintervention because of malposition or spontaneous extrusion of the gold weight. It was removed in 4 of them. Upon closure the average residual palpebral aperture was 1.17 mm (range: 0 5) in the patients with gold weight in situ and 6.00 mm (5-10) in the patients in whom the implant had been removed. CONCLUSION: A better eye closure was obtained in 82% of the patients, while eye complaints resolved in 36% of them. PMID- 10800551 TI - [Pregnancy in women with diabetes mellitus type I: maternal and perinatal complications, in spite of good blood glucose control]. AB - OBJECTIVE: To examine the maternal and neonatal outcome of pregnancies of women with type I diabetes mellitus. DESIGN: Retrospective. METHODS: The medical records of pregnancies (> or = 16 weeks) in women with type I diabetes mellitus between 1986/'97 were studied in University Medical Center Utrecht, Academic Hospital Groningen and Isala Clinics, location 'De Weezenlanden', Zwolle, the Netherlands. RESULTS: During the study period, 172 women had 220 pregnancies: 212 single and 8 twin pregnancies. The mean age was 29.1 years (SD: 4.1), the mean duration of standing of the diabetes was 12 years (range: 1-32) and the mean concentration of glycosylated haemoglobin (HbA1c) was 6.3% at 10 weeks of pregnancy. The incidence of children with congenital malformations was 4 times higher (n = 19; 9.0%) than that in the Dutch population (2%). Macrosomia occurred in 92 children (43.4%) and perinatal mortality in 7 (3.3%). Maternal hypertensive complications occurred in 39 single pregnancies (18.4%), which is 2-3 times more often than in the Dutch population. CONCLUSION: In type I diabetic women maternal complications, perinatal morbidity and mortality are increased, despite near optimal glycaemic control. PMID- 10800552 TI - [Periodic fever due to hyper-IgD syndrome]. AB - In a 45-year-old man who from early childhood had been suffering of periodic fever, which did not respond to any therapy attempted, the ultimate diagnosis was hyperimmunoglobulinaemia D syndrome (HIDS). HIDS attacks typically occur every 4 6 weeks and last 3-7 days. The most frequent symptoms are fever, diarrhoea, arthralgias, cold shivers, abdominal pain, vomiting and headache. Physical examination often reveals lymphadenopathy, skin lesions, arthritides, splenomegaly and serositis. Laboratory investigation includes an acute-phase response with granulocytosis and enhanced erythrocyte sedimentation rate. The serum concentration of IgD is increased as is the concentration of IgA. There is no causal therapy. A causative gene mutation was recently identified. PMID- 10800553 TI - [Drug interactions of Hypericum perforatum (St. John's wort) are potentially hazardous]. AB - Hypericum can lower the plasma levels of simultaneously administered drugs by induction of metabolism. Combinations of hypericum products with warfarin, cyclosporin, oral contraceptives, theophylline, fenprocoumon, digoxin and indinavir have led to reported interactions and reduced therapeutic activity. It is therefore not advisable to combine hypericum products with other drugs, especially CYP3A4 and p-glycoprotein substrates. Discontinuing hypericum after protracted use may lead to higher plasma levels of the drugs used simultaneously, with the risk of adverse effects. Registered homeopathic preparations with a dilution of 1 in 10,000 or weaker may be regarded as safe. PMID- 10800554 TI - [Successful treatment of an elderly woman after stubborn resistance]. PMID- 10800555 TI - [Revised CBO guideline 'Urinary tract infections']. PMID- 10800556 TI - [Additional effects of statins independent of the cholesterol-lowering as yet not shown to be clinically relevant]. PMID- 10800557 TI - [National Cancer Fund (Koningin Wilhelmina Fonds) and Houtsmuller therapy in cancer]. PMID- 10800558 TI - [Sinus cavernosus syndrome]. PMID- 10800559 TI - [Obituary to Artur Galanzka, physician epidemiologist 1933-1999]. PMID- 10800560 TI - [Professor Jozef Karol Kostrzewski (Jan. 22. 1883-Apr. 5.1959)]. PMID- 10800562 TI - [The concept of herd immunity applied to the evaluation of vaccination programs]. AB - The article concerns the definition and ways of estimation of indicators of herd immunity as applied to the analysis of the efficiency of vaccination programmes. Some important factors are discussed which determine the proportion of vaccinated individuals in the population necessary for decrease of incidence and eventually for elimination of the disease from a population. The problems related to definition and calculation of herd immunity in populations non uniform in terms of individual risk of infection are discussed. PMID- 10800561 TI - [Chlamydia pneumoniae: an etiologic of coronary heart disease?]. AB - The hypothesis put forward in 1988 that Chlamydia pneumoniae is the aetiological agent in coronary disease and myocardial infraction has aroused an interest in these bacteria. The epidemiology of Ch. pneumoniae infections and researches on the role of it in the development of coronary artery lesions are reviewed, including animal models of this infection which could provide additional on the mechanism of atherosclerosis development. PMID- 10800563 TI - [What do we mean by the concept of principles of investigation?]. AB - It is a review of basic epidemiological techniques used for investigation of an outbreak of epidemic. The author defines types of epidemics and characterizes conditions of their occurrence and progression in spatial and temporal ramifications. The basic types of epidemiological research used in epidemiological analysis such as cohort and case-control study are characterized and exemplified with application to the outbreak of epidemic. The article provides in the conclusion basic logistic and organizational principles for such an investigation. PMID- 10800564 TI - [Introduction to the epidemiology of disease caused by Haemophilus influenzae]. AB - This review article characterizes Haemophilus influenzae bacteria and presents basic clinical syndromes caused by it. It informs about sources and pathways of Haemophilus influenzae infection and gives basic comparative data on its epidemiology in different countries, and also different ethnic and social groups. PMID- 10800565 TI - [Measles vaccines]. AB - The World Health Organization has settled the goal of measles elimination in European Region by the year 2007. The proposed measles elimination strategy is to reduce an estimated proportion of susceptible individuals in the population to low levels by intensive vaccination and to maintain these low levels for some years. In this strategy, the quality of measles vaccine used for immunization is crucial. The different kinds of measles vaccines used at present in the world and novel generations of vaccines are presented. PMID- 10800566 TI - [Serological survey of measles antibody prevalence in chosen regions in Poland]. AB - Serological survey of measles antibody prevalence in the Polish population was carried out in 1998. The study group consisted of population in 6 regions: Warsaw, Wroclaw, Poznan, Lublin, Tarnobrzeg and Zielona Gora. A total of 3,000 serum samples were collected from individuals aged from 12 months to 30 years. Sera were assayed for measles specific IgG by ELISA test. All samples with titres < or = 0.5 IU were considered as negative. Serological survey has revealed a total of 4% negative samples. The highest percentage of susceptibles was noted among persons aged 15-19 years (8.2%), and among children who have just finished their first year of life (8%). The need for revaccination of teenagers aged 15-19 years is suggested. PMID- 10800567 TI - [Ticks Ixodes ricinus (Linnaeus, 1758) from selected municipal forests of the city Poznan and their infection with the spirochetes Borrelia burgdorferi senso lato]. AB - The results of research (1997-1998) on the occurrence of the spirochetes Borrelia burgdorferi s.l. in ticks Ixodes ricinus (L.) in selected municipal forests of the city of Poznan (localities--districts of Poznan: 1 Debiec, 2 Marcelin, 3 Golecin and Wola, 4 Krzyzowniki and Smochowice, 5 Kiekrz and Strzeszynek, 6 Morasko, 7 Piatkowo, 8 Umultowo, 9 Naramowice and Rozany Mlyn, 10 Malta and Antoninek) are presented. A total of 1432 ticks were collected from 10 localities (748 larvae, 590 nymphs, 47 males, and 47 females). Out of this number, 266 specimens were selected at random for further analysis (20 larvae, 160 nymphs, 44 males, and 42 females) which were then tested for the presence of the spirochetes using the PCR technique (tab. I-III, fig. 1). Spirochetes were found in 60 specimens (22.6%). Percentages of infected nymphs and males were similar: 25.6% and 27.3%, respectively. The level of infection of females was lower (14.3%) and of larvae lowest (5.0%). Infected ticks were found in all ten localities, but their proportions differed from site to site and varied from 9.5% (Piatkowo) to 34.6% (Krzyzowniki and Smochowice). Special attention was devoted to those municipal woods where ticks were abundant (Kiekrz and Strzeszynek, as well as Malta and Antoninek, and relatively numerous Debiec) and where their extensiveness of infestation was high (Krzyzowniki and Smochowice: 34.6%; Debiec: 29.6%; Naramowice and Rozany Mlyn: 28.6%; Malta and Antoninek: 28.1%; and Golecin and Wola: 26.9%). After summing up all the data from the literature on Wielkopolska and the city of Poznan, the mean extensiveness of infestation in Wielkopolska turns out to be 21.8% and in Poznan slightly higher, 22.2% (tab. IV). The infection levels of the particular developmental stages are different, though: highly variable in nymphs (8.3-25.6%), and more stable in males (22.0 27.3%) and females (22.0-24.5%). PMID- 10800568 TI - [Anthrax in man with reference to alimentary infection and environmental conditions]. AB - Based on already published data, the following issues have been covered in the present review: epidemiology, epizootiology and the occurrence of anthrax in man with reference to current data. Moreover, in the paper are presented some problems of intestinal anthrax with emphasis on environment conditions, pathogenesis, prevention and treatment of this clinical form. PMID- 10800569 TI - [Performance of vaccinations against poliomyelitis in the province of Krakow versus the plan of elimination and eradication of this disease in Poland]. AB - The criteria of elimination and eradication of poliomyelitis set down by the WHO and the performance of vaccinations are presented in the Province of Cracow and are compared with the plan of the National Programme of Elimination and Eradication of Poliomyelitis. The analysed period covered the years 1991-1998. In 1998 the World Health Congress passed the resolution on the elimination and eradication of poliomyelitis worldwide. For accepting eradication achieved the criterion is required of vaccination of 90-95% of babies aged under 2 years in the whole country, and not less than 90% in individual provinces. In early 1990's below 75% babies in that age group were vaccinated in the Province of Cracow. Only in 1998 for the first time the 92% rate of vaccinations as required by the WHO was achieved. PMID- 10800570 TI - [Lyme Borreliosis]. AB - The epidemiology of Lyme borreliosis is described with stress laid on the relations between ticks as vectors and the animals which are the reservoir of borrelia infecting man. Other discussed problems include the possibility of infection of man, the seasonal character of new cases, the worldwide presence of Ixodes ticks and their species in various continents, the incidence of Lyme borreliosis in various countries. PMID- 10800571 TI - [The occurrence of intestinal parasites among children attending first classes of the elementary schools in Poland in the school year 1997/1998]. AB - OBJECTIVE: The studies were undertaken to: (i) evaluate the epidemiological situation of infections with intestinal parasites among seven-year-old children attending first classes of the elementary schools in 25 provinces of Poland (ii) answer the question concerning an influence of the summer flood 1997 on the epidemiological situation of intestinal parasitoses in the affected regions. METHODS: The studies included 30,110 children, i.e. 10% of the population of seven-year-olds, in the provinces under study, and were based on the examination of a single faecal specimen and a cellophane swab using the following methods: wet mount preparations in 0.9% NaCl and diluted Lugol,s iodine, zinc sulphate centrifugal flotation and decantation. RESULTS AND CONCLUSIONS: The parasites most frequently encountered in the examinations included: Enterobius vermicularis (in 16.45% of the examined), Ascaris lumbricoides (2.8%), Giardia intestinalis (1.02%), Entamoeba coli (0.73%), Trichuris trichiura (0.29%). The obtained results confirmed the decreasing frequency of infections with intestinal parasites in Poland and indicated that the summer flood of 1997 had a little influence on the epidemiological situation of intestinal parasitoses in the affected regions. PMID- 10800572 TI - [The frequency of giardiasis in various children's environment]. AB - Giardiasis is an intestinal parastic disease, occurring in many world regions. Its incidence is variable and depends on such factors as studied population and diagnostic methods. In this study the incidence of Giardiasis was assessed in children living in various environments and living conditions. Stool examination was made using microscopic and immunoenzymatic methods. Among 208 examined children 42 (20.2%) were infected with Giardia lamblia. Giardiasis was the most common in children living in orphanage--41.4%. Incidence of Giardiasis among children from day--care centers was the lowest--3.7%. Children aged 12-14 years were infected in 42.7% and aged 0-2 years only in 12.5%. There was no significant difference between boys and girls. PMID- 10800573 TI - [Cryptosporidium parvum: an emerging threat to the man]. AB - The current opinions on the biology, epidemiology and potential role of Cryptosporidium parvum as an emerging, highly infectious threat are presented. Available Polish data concerning the frequency of cryptosporidiosis were gathered and the diagnostic procedures, pathogenesis and treatment of cryptosporidiosis are described. PMID- 10800574 TI - [Salmonella serovars defined in Poland]. AB - One hundred and sixty nine serovars of Salmonella, belonging to 28 different O groups, were defined in the National Salmonella Centre. All the serovars were isolated in Poland. Above 70% (121/169) of them belonged to five O groups--O:4 (31 serovars), O:7 (28 serovars), O:8 (27 serovars), O:3,10 (20 serovars) and O:9 (15 serovars). Other O groups were represented by some (1 to 8) serovars. All the serovars were described according to the changes in taxonomy of the genus Salmonella and in serovars nomenclature. These changes, proposed by WHO Collaborating Centre for Reference and Research on Salmonella (Paris, France), reflect the recent advances in Salmonella taxonomy. PMID- 10800575 TI - [Salmonella serovars determined in the National Salmonella Center between 1995 and 1997]. AB - A total of 897 human and non-human (from animal, food, feed and other sources) isolates of Salmonella were serotyped in the National Salmonella Centre during the period 1995-1997. They had been referred to the Centre by several field laboratories all over the country. Serological and biochemical identification was performed according to the standard methods. Forty seven Salmonella serovars were isolated from human sources. The most common Salmonella serovars were S. Thompson, S. Mbandaka and S. Enteritidis. The other were S. Hadar, S. Infantis, S. Albany, S. Typhimurium and S. Virchow. The serovars isolated from the food products were almost the same as from human sources. A total of 37 Salmonella serovars were found in isolates from animal sources. The dominating serovars were S. Enteritidis, S. Mbandaka, S. Bredeney, S. Infantis, S. Anatum, S. Typhimurium, S. Hadar, S. Saintpaul, S. Agona, S. Virchow, S. Heidelberg and S. Thompson. The fact that a number of the same serovars were found on lists of the most commonly isolated serovars from human and animal sources underlines the importance of the relationship. The lack of information of Salmonella contaminated food products and animal infections was noticed More detail examination of isolates from animal sources is advised to help the fight against human salmonellosis. PMID- 10800576 TI - [AIDS syndrome in an eight-month-old infant]. AB - The paper presents multiorgan manifestations of AIDS syndrome in an infant at the age of 8 months. The child was admitted to the Clinic with enteric disorder, anaemia, hepatosplenomegaly and pneumonia. The diagnosis of those anomalies and the treatment of pneumonia took much time. Infection with CMV was recognized but, despite the treatment and elimination of the virus, the child's condition did not improve and general emaciation progressed. The diagnosis of AIDS syndrome was based upon indicator illnesses: chronic recurrent pneumonia, cytomegaly and emaciation syndrome as well as upon the results of additional examinations, first of all including the presence of p24 antigen in the serum. In children with chronic diseases and manifesting non-specific multiorgan symptoms we should take into consideration AIDS syndrome in the differential diagnosis. PMID- 10800577 TI - [Characteristic of verotoxigenic strains of Escherichia coli]. AB - The aim of the study was the isolation from faecal samples of patients with diarrhoea of verotoxigenic strains of E. coli (VTEC) on the basis of characteristic biochemical properties and production of enterohaemolysin and comparison of isolated verotoxigenic strains with reference strains of VTEC. For isolation of VTEC from 257 stool samples derived from patients with diarrhoea were used selective medium sorbiol--Mac Conkey agar (SMAC) and media supplemented with unwashed and washed in PBS sheep erythrocytes for detection of haemolysins of E. coli. In all haemolytic and sorbitolo-positive or -negative strains isolated from 93 stool samples were examined the activity of beta-glucuronidase using MUG (4-methylumbelliferyl-beta-D-glukuronid) as a substrate for that enzyme. All isolated haemolytic strains as well as reference VTEC were examined on Vero cell line. Verotoxigenic strains from examined samples were investigated by agglutination assay with antiserum to E. coli O157 and then with antisera to eneropathogenic E. coli (EPEC). After that they were examined with ID GN and ATB GN tests. In 93 (36.2%) examined samples there were haemolytic strains of E. coli which fermented or not sorbitol and were MUG-positive or negative. Only in 2 (0.2%) stool samples there were verotoxigenic strains of E. coli which were sorbiol-positive and MG-positive. Both strains belonged to O26 serotype and were derived from samples of two children with diarrhoea. Isolated verotoxigenic strains of E. coli O26 were susceptible on all tested antibiotics. PMID- 10800578 TI - [The HLA-I and HLA-II expression evaluation in liver biopsy specimens of patients with chronic viral hepatitis]. AB - The main an etiological agents of chronic hepatitis are viral infections. The viral infection course and outcome depend mostly on the immunological response. Infected hepatocytes are damaged by appropriately viral antigen-specific cytolytic T lymphocytes. Those sensitised T cells react only with those hepatocytes which express viral antigen and antigen HLA on membrane surface. The aim of this study was to evaluate the expression of selected histocompatibility antigens HLA in liver biopsy specimens of patients with chronic viral hepatitis. Seventeen patients with chronic persistent hepatitis (inflammatory activity 1-4 points according to Scheuer scale modified by Gabriel) and 27 patients with chronic active hepatitis (5-10 points) were studied. In these groups of patients the intensity of HLA-I (A, B, C), HLA-II (DR) expression in liver biopsy specimens, alanine aminotransferase activity, markers of HBV and HCV in serum were examined. The monoclonal mouse anti-human antibodies and streptavidin-biotin with alkaline phosphatase method for estimation of HLA-I, HLA-II was used. Results were statistically analysed using Mann-Whitney's U test and Spearman's rank correlation test. Generally, the expression of HLA-I and HLA-II on hepatocyte membrane was shown. Significant differences in expression of HLA-II among studied groups were observed, moreover the highest degree of HLA-II intensity in the group of patients with greater inflammation activity was significantly more frequently observed. The expression of HLA-I, HLA-II was regardless of the viral a etiology and serological markers of HBV replication. The degree of studied parameters expression was positively correlated with biochemical activity of inflammation. PMID- 10800579 TI - [Protective vaccinations in prevention of acute infectious diseases in Poland in 1918-1923]. AB - The aim of the paper is to demonstrate the role of protective vaccinations, including obligatory ones and the so called vaccinations in emergency, in control of infectious diseases in the years 1918-1923. The legal regulations of protective vaccinations the supply of vaccines and the organization and performance of obligatory vaccinations against smallpox, typhoid fever, paratyphus A and B, and Asiatic cholera made possible, together with other preventive measures, to restrict and eradicate foci of infections. PMID- 10800580 TI - [Primary coronary angioplasty in acute myocardial infarction]. AB - It is well established that recanalization of the infarct-related artery has an important role in prognosis after myocardial infarction. Recanalization can be achieved by the use of thrombolytic agents or by percutaneous transluminal coronary angioplasty. This article reviews the role of primary coronary angioplasty in the management of acute myocardial infarction, and summarises current opinions and indications of its use. PMID- 10800582 TI - [A comparison of myocardial infarction clinical course and long-term prognosis in the years 1977-85 and 1992-96]. AB - This study describes comparison of the management in 997 patients treated in 1977 85 with acute transmural myocardial infarction (MI), their in-hospital and long term mortality, with 881 patients treated in 1992-96. The study included geographically the same population treated by the same medical staff but in another decade period and the same MI criteria in both cohorts. Main changes were associated with the management of MI. Both groups were adjusted to age (59.3 +/- 11 vs. 59.9 +/- 11.7), history prior to myocardial infarction, diabetes mellitus, habit of smoking and hospital complications except rarer presence of block A-V II degrees and III degrees in the period 1992-96. The increase in percentage of female patients was noticed in the analysis (from 21% to 28.5%). The crude in hospital mortality was statistically lower in 1992-96 than in 1977-85 (19.8% vs. 13.4% p < 0.11) and the follow up mortality in the next 2-6 years decreased from 6.97% to 4.95%. Mortality in the first year after MI in both periods was high and raised to 9.6% and 8.9%, respectively (NS). Significant diminish of mortality in next years after myocardial infarction was observed in all age adjusted groups of patients treated in 1992-96. PMID- 10800581 TI - [The effect of short-term changes in levels of air pollution on mortality from cardiovascular diseases among inhabitants of Krakow]. AB - The analysis of short term relation between daily ambient air pollution and daily number of deaths in Krakow, Poland, during the period 1993-1996 was the purpose of the study. The exposure time series data dealt with sulphur dioxide (SO2) and particles with aero-diameter smaller than 10 microns (PM10). Daily mortality due to all causes and cardiovascular diseases were considered separately for two age groups. The statistical procedures included modelling of potential confounding factors (seasonal patterns, meteorological factors) and in the final analysis the Poisson regression model was applied. Effects were expressed as relative risks per 100 micrograms/m3 increase of the corresponding pollutant levels. It was found that sulphur dioxide was significantly related to mortality from all causes and cardiovascular conditions in the age group 65 years and over whereas the effect of suspended particles was at the borderline significance level. The relative risk of cardiovascular death associated with 100 micrograms/m3 increment of mean daily SO2 was 1.17 (1.10-1.25) in the total sample under study, higher for men (RR = 1.27, CI: 1.12-1.45) than for women (RR = 1.12, CI: 1.01-1.23). The corresponding RRs for PM10 and cardiovascular deaths were 1.06 (0.98-1.16) for men and 1.07 (0.99-1.13) for women. These results strengthen the evidence of casual relationship between ambient air pollution level and daily mortality and pinpoints important health hazards issues for the Krakow inhabitants. PMID- 10800583 TI - [Analysis of reasons for discontinuation of treatment for dyslipidemia in secondary prevention of coronary heart disease]. AB - Objective of the study was retrospective assessment of the reasons of discontinuing the treatment of dyslipidaemias in patients with confirmed coronary heart disease. The study wa on 100 patients (70 M, 30 F, age 30-74 yrs) treated in the Cardiological Center of Ambulatory Diagnostics and Rehabilitation in Wroclaw from 01.01.1995 to 01.03.1998. It was shown that the average period of the treatment in Cardiological Center lasted 17.9 months (in the patients who stopped the therapy 13.1 months) and it was not correlated with sex, type of the hyperlipidaemia, the severity of the disease and with type and dosage of the drugs. Cardiosurgical treatment of the coronary heart disease and the age of the patient were the factors correlated significantly with the average period of the lipid-lowering therapy. The absence of a strong (created by the physician) motivation in the patient, and also the deficiency of understanding and his aware agreement for the continuous therapy of the asymptomatic disease, was the main reason of the failure. PMID- 10800584 TI - [The influence of angiotensin-converting enzyme inhibitors on collagen content of the aorta wall in experimental hypercholesterolemia]. AB - In atherosclerosis numerous qualitative and quantitative changes in connective tissue metabolism parameters in serum and aorta occur. In atherosclerosis there is an enhanced activity of local renin-angiotensin systems. It leads to overexpression of ANG II, both in serum and arterial wall. ANG II stimulates SMC to over-synthesize the collagens type I and III. Hyper-cholesterolemia is a form of metabolic injury which can both induce phenotypic change of SMC and activate RA system in arterial wall. ACEI lower the accumulation of collagens type I and III, and enhance elastin content in arterial wall in experimental hypertension. The aim of this study was to assess the influence of captopril, enalapril and quinapril on connective tissue metabolism of the aorta in experimental hyper cholesterolemia. 64 male New Zealand rabbits were used. Animals were fed with standard fodder, special diet (1% cholesterol content) or special diet + tested ACEI. Two doses of ACE inhibitors were used: 1st--equivalent to doses applied to human subjects (in mg/kg of body weight), 2nd--dose 10 times higher. The animals were divided into 8 equal groups: K--standard fodder, B--special diet, C1, C2- special diet + captopril in doses 2.5 and 25 mg/kg/24 hours, respectively, E1, E2 -special diet + enalapril in doses 0.75 and 7.5 mg/kg/24 hours, respectively, Q1 i Q2--special diet + quinapril in doses 0.75 and 7.5 mg/kg per day, respectively. The experiment lasted for 6 months. After 24 weeks the animals were sacrificed and aortae were excised for collagens assay. The statistical analysis was performed using ANOVA, followed by LSD test; p < 0.05 was considered statistically significant. The aorta collagens content of cholesterol-fed rabbits significantly increased. The tested ACEI (captopril, enalapril in both doses and quinapril in lower dose) had a preventive effect against the increase of aorta collagen content. PMID- 10800585 TI - [Neuropsychological disorders after subarachnoid hemorrhage and surgery in anterior communicating artery brain aneurysms]. AB - In the years 1982-1985 in the Dept. of Neurosurgery of the Jagiellonian University, 197 patients with anterior communicating artery aneurysms were operated on. Excluding multiple aneurysms (9), and taking into account early surgical losses (16), 172 patients were supposed to undergo late medical examinations. The patients were examined by means of questionnaires at least five years after surgery. 43 patients out of the 123 (71.5%) who handed in filled-in questionnaires, underwent psychological examination. Preliminary analysis of the late results of surgery indicates maintenance of the neurological disorders and symptoms. Within the group of examined patients (a neuropsychological examination, Wechsler Scale) it was found that 33 patients had considerable morbid decline in their mental condition and 4 patients moderate. 6 patients were not examined with Wechsler Scale because of aphasia (5 patients) and considerable deterioration of visual acquity (1 patient). The results of the neuropsychological examination showed that the symptoms that predominated were: short memory disorder, a slowdown in psychical and motor functions, conscious concentration disorder, elements of dynamic apraxia and increased tiredness as far as psychical process was concerned. 14 patients (32.6%) resumed work on their former posts, among them the 4 patients who had moderate morbid decline in their mental condition. PMID- 10800586 TI - [Use of polymerase chain reaction in medical diagnosis]. AB - Principles of the polymerase chain reaction (PCR) and its modifications are discussed: reverse transcription-PCR (RT-PCR), nested PCR, restriction fragment length polymorphism-PCR (RFLP-PCR). Different methods of the PCR products detection used in routine diagnostics and research are presented: electrophoresis and chromatography techniques, immunoenzymatic methods, hybridisation and sequencing. Applications of PCR reaction in diagnostics of infectious diseases (viral and bacterial), genetic diseases and genotyping in transplantation are discussed and examples of commercial tests based on PCR reaction and another molecular biology methods used in fast, routine diagnostics are listed. PMID- 10800587 TI - [Hereditary inclusion-body myopathy]. AB - Hereditary inclusion-body myopathy (h-IBM) is a heterogeneous group of disorders characterized by progressive weakness of some skeletal muscles and pathological feature, intercellular rimmed vacuoles in the muscular filaments. h-IBM is not associated with inflammatory infiltrations and term myopathy is used as opposite to spontaneous inclusion-body myositis. h-IBM is classified into autosomal recessive and autosomal dominant subgroups. PMID- 10800588 TI - [Controversial opinions regarding the role of Bcml vitamin D receptor gene polymorphism on parathyroid function, calcium absorption and bone mineral density in persons with normal renal function and patients with chronic renal failure]. AB - This paper summarizes current, but controversial opinions of different authors evaluating the effect of Bsml polymorphism of the vitamin D receptor gene on the parathyroid function, calcium absorption from the digestive tract and bone mineral density (BMD) in persons with normal renal function and in patients with chronic renal failure. On the basis of these data, the recapitulation of the prevailing opinions was undertaken. Contemporary it is admitted, that the persons with bb genotype and normal renal function are at risk for the development of the parathyroid adenomas while in the patients with chronic renal failure the bb genotype may favour parathyroids hyperplasia. It is also believed that bb genotype in the persons with normal renal function facilitates calcium absorption and is associated with greater BMD. Few investigations performed in the patients with chronic renal failure suggest that the BB genotype may partially influence lower BMD in the subgroup of younger patients treated with hemodialysis for a relatively short period. Existing controversies need however further clarification. PMID- 10800589 TI - [A case of pseudohypoparathyroidism]. AB - We presented a case of a 22-year-old woman with pseudohypoparathyroidism type la. She was a typical congenital Albright syndrome patient with osteodystrophy including hands and feet, accompanied by obesity, strabismum, and retardation growth. Her calcium and phosphate levels were within the normal range which was stated during repeated hospitalisation due to infections. The diagnosis of PHPT was made at the age of 22 when the patient suffered twice from tetany seizure accompanied by numbness and tingling sensation in her hands and around the mouth as well as cramps in her legs. Typical phenotype were found: shortness in stature, obesity, rounded face, small hands and shortening of the third, the fourth and the fifth fingers in both hands and the third and the fourth toes in feet together with trophic disorders of nails and valgity of her knees. Somatic sings were accompanied by hypocalcaemia and hyperphosphatemia, hyperphosphatasia, lower calcium and phosphate urinary excretion and three-fold increased PTH serum level. Computerised tomography of the brain showed extensive evidence of cerebral calcification in basal ganglia as well as in dura mater and in skin covering the skull. X-ray and densitometry examinations revealed osteolytic foci in cranial, humeral and forearm bones as well as osteoporosis in palm and feet bones. The patient presented a typical case of PHPT with resistance of the kidney to PTH, what was confirmed by lower calcium and phosphate urinary excretion, with normal bone receptor sensibility to PTH. Elevated PTH levels resulted in osteoporosis and foci of osteolysis. Treatment with calcium and active form of vitamin D3 caused reversal of hypocalcemia symptoms and normalisation of biochemical features. We also found hormonal symptoms of latent hypothyreosis. No coexistence of PTH with thyroid receptor resistance was found. The case was described because it is rare disorder, difficult to diagnose. Early diagnosis and treatment is necessary to limit the irreversible changes as well as bone and central nervous system injury. PMID- 10800590 TI - [Neuromas of the extrahepatic bile ducts]. AB - Authors collected three cases of neuromas situated in the wall of extrahepatic bile ducts, which occluded lumen and caused jaundice. In two cases without any evidence of previous surgery or cholelithiasis, neuroma was localized in ductus choledochus. After excision of ductus choledochus and cholecystectomy, we performed hepaticojejunostomy Roux--en Y. In the last patient, 7 years after cholecystectomy, neuroma localized in hepatic duct was treated by T-tube. In all patients with mechanical jaundice, before operation we had never correctly diagnosed this very rare cause of obstruction of extrahepatic bile ducts. PMID- 10800591 TI - [About word meaning in medicine--deliberation about language]. PMID- 10800592 TI - Heterotrophic assimilation of carbon dioxide. 1942. PMID- 10800593 TI - Phosphotriesterase: an enzyme in search of its natural substrate. AB - The bacterial PTE is able to catalyze the hydrolysis of a wide range of organophosphate nerve agents. The active site has been shown to consist of a unique binuclear metal center that has evolved to deliver hydroxide to the site of bond cleavage. The reaction rate for the hydrolysis of activated substrates such as paraoxon is limited by product release or an associated protein conformational change. PMID- 10800594 TI - Phosphoribulokinase: current perspectives on the structure/function basis for regulation and catalysis. AB - Phosphoribulokinase (PRK), an enzyme unique to the reductive pentose phosphate pathway of CO2 assimilation, exhibits distinctive contrasting properties when the proteins from eukaryotic and prokaryotic sources are compared. The eukaryotic PRKs are typically dimers of -39 kDa subunits while the prokaryotic PRKs are octamers of -32 kDa subunits. The enzymes from these two classes are regulated by different mechanisms. Thioredoxin of mediated thiol-disulfide exchange interconverts eukaryotic PRKs between reduced (active) and oxidized (inactive) forms. Allosteric effectors, including activator NADH and inhibitors AMP and phosphoenolpyruvate, regulate activity of prokaryotic PRK. The effector binding site has been identified in the high resolution structure recently elucidated for prokaryotic PRK and the7 apparatus for transmission of the allosteric stimulus has been identified. Additional contrasts between PRKs include marked differences in primary structure between eukaryotic and prokaryotic PRKs. Alignment of all available deduced PRK sequences indicates that less than 10% of the amino acid residues are invariant. In contrast to these differences, the mechanism for ribulose 1,5-biphosphate synthesis from ATP and ribulose 5-phosphate (Ru5P) appears to be the same for all PRKs. Consensus sequences associated with M++-ATP binding, identified in all PRK proteins, are closely juxtaposed to the residue proposed to function as general base catalyst. Sequence homology and mutagenesis approaches have suggested several residues that may potentially function in Ru5P binding. Not all of these proposed Ru5P binding residues are closely juxtaposed in the structure of unliganded PRK. Mechanistic approaches have been employed to investigate the amino acids which influence K(m Ru5P) and identify those amino acids most directly involved in Ru5P binding. PRK is one member of a family of phospho or sulfo transferase proteins which exhibit a nucleotide monophosphate kinase fold. Structure/function correlations elucidated for PRK suggest analogous assignments for other members of this family of proteins. PMID- 10800595 TI - The molecular evolution of pyridoxal-5'-phosphate-dependent enzymes. AB - The pyridoxal-5-phosphate-dependent enzymes (B6 enzymes) that act on amino acid substrates are of multiple evolutionary origin. The numerous common mechanistic features of B6 enzymes thus are not historical traits passed on from a common ancestor enzyme but rather reflect evolutionary or chemical necessities. Family profile analysis of amino acid sequences supported by comparison of the available three-dimensional (3-D) crystal structures indicates that the B6 enzymes known to date belong to four independent evolutionary lineages of homologous (or more precisely paralogous) proteins, of which the alpha family is by far the largest. The alpha family (with aspartate aminotransferase as the prototype enzyme) includes enzymes that catalyze, with several exceptions, transformations of amino acids in which the covalency changes are limited to the same carbon atom that carries the amino group forming the imine linkage with the coenzyme (i.e., Calpha in most cases). Enzymes of the beta family (tryptophan synthase beta as the prototype enzyme) mainly catalyze replacement and elimination reactions at Cbeta. The D-alanine aminotransferase family and the alanine racemase family are the two other independent lineages, both with relatively few member enzymes. The primordial pyridoxal-5-phosphate-dependent enzymes apparently were regio-specific catalysts that first diverged into reaction-specific enzymes and then specialized for substrate specificity. Aminotransferases as well as amino acid decarboxylases are found in two different evolutionary lineages. Comparison of sequences from eukaryotic, archebacterial, and eubacterial species indicates that the functional specialization of most B6 enzymes has occurred already in the universal ancestor cell. The cofactor pyridoxal-5-phosphate must have emerged very early in biological evolution; conceivably, organic cofactors and metal ions were the first biological catalysts. In attempts to stimulate particular steps of molecular evolution, oligonucleotide-directed mutagenesis of active-site residues and directed molecular evolution have been applied to change both the substrate and reaction specificity of existent B6 enzymes. Pyridoxal-5-phosphate-dependent catalytic antibodies were elicited with a screening protocol that applied functional selection criteria as they might have been operative in the evolution of protein-assisted pyridoxal catalysis. PMID- 10800596 TI - O-acetylserine sulfhydrylase. AB - The 31P NMR data suggest slight differences in the structures around the 5'-P for the internal Schiff base and the lanthionine external Schiff base (both largely ketoeneamine) and a large difference for enolimine portion of the serine external Schiff base. Addition of cysteine or serine increase delayed fluorescence and triplet to singlet energy transfer. Addition of OAS exhibits a splitting of the 0,0 vibronic, the result of two distinct conformations, likely enolimine and ketoeneamine tautomers. Nonetheless, the alpha-amino-acrylate Schiff base conformation differs from either the internal or external Schiff base conformations. All of the time-resolved fluorescence data are consistent with conformation changes reflecting redistribution of ketoeneamine and enolimine tautomers as catalysis occurs. It is important to remember that the structural changes are substantial. The native structure (internal Schiff base) is active site open, while the K41A mutant enzyme (ketoeneamine external Schiff base) is active site closed. The trigger for the conformational change from open to closed as one goes from the internal to external Schiff base is the occupancy of the alpha-carboxyl subsite of the active site (Burkhard et al., 1999). Associated with this, as observed in pH-rate profiles, pH-dependent changes in phosphorescence, and pH-dependent changes in fluorescence enhancement upon binding acetate or cysteine is an enzyme group with a pK in the range 7-8. Dependent on the protonation state of the enzyme group, structural changes likely occur that also reflect a redistribution of the tautomeric equilibrium. Finally, the minimal catalytic cycle can likely be pictured as shown in Fig. 20. The changes may be pH dependent, and the open conformations for the internal Schiff base and the alpha-aminoacrylate Schiff base are not identical structurally, as expected because of the increased stability of the latter. PMID- 10800597 TI - The aromatic amino acid hydroxylases. AB - The enzymes phenylalanine hydroxylase, tyrosine hydroxylase, and tryptophan hydroxylase constitute the family of pterin-dependent aromatic amino acid hydroxylases. Each enzyme catalyzes the hydroxylation of the aromatic side chain of its respective amino acid substrate using molecular oxygen and a tetrahydropterin as substrates. Recent advances have provided insights into the structures, mechanisms, and regulation of these enzymes. The eukaryotic enzymes are homotetramers comprised of homologous catalytic domains and discrete regulatory domains. The ligands to the active site iron atom as well as residues involved in substrate binding have been identified from a combination of structural studies and site-directed mutagenesis. Mechanistic studies with nonphysiological and isotopically substituted substrates have provided details of the mechanism of hydroxylation. While the complex regulatory properties of phenylalanine and tyrosine hydroxylase are still not fully understood, effects of regulation on key kinetic parameters have been identified. Phenylalanine hydroxylase is regulated by an interaction between phosphorylation and allosteric regulation by substrates. Tyrosine hydroxylase is regulated by phosphorylation and feedback inhibition by catecholamines. PMID- 10800598 TI - L-aspartase: new tricks from an old enzyme. AB - The enzyme L-aspartate ammonia-lyase (aspartase) catalyzes the reversible deamination of the amino acid L-aspartic acid, using a carbanion mechanism to produce fumaric acid and ammonium ion. Aspartase is among the most specific enzymes known with extensive studies failing, until recently, to identify any alternative amino acid substrates that can replace L-aspartic acid. Aspartases from different organisms show high sequence homology, and this homology extends to functionally related enzymes such as the class II fumarases, the argininosuccinate and adenylosuccinate lyases. The high-resolution structure of aspartase reveals a monomer that is composed of three domains oriented in an elongated S-shape. The central domain, comprised of five-helices, provides the subunit contacts in the functionally active tetramer. The active sites are located in clefts between the subunits and structural and mutagenic studies have identified several of the active site functional groups. While the catalytic activity of this enzyme has been known for nearly 100 years, a number of recent studies have revealed some interesting and unexpected new properties of this reasonably well-characterized enzyme. The non-linear kinetics that are seen under certain conditions have been shown to be caused by the presence of a separate regulatory site. The substrate, aspartic acid, can also play the role of an activator, binding at this site along with a required divalent metal ion. Truncation of the carboxyl terminus of aspartase at specific positions leads to an enhancement of the catalytic activity of the enzyme. Truncations in this region also have been found to introduce a new, non-enzymatic biological activity into aspartase, the ability to specifically enhance the activation of plasminogen to plasmin by tissue plasminogen activator. Even after a century of investigation there are clearly a number of aspects of this multifaceted enzyme that remain to be explored. PMID- 10800599 TI - One size fits all? PMID- 10800600 TI - Hemostatic defects in massive transfusion: rapid diagnosis and management. AB - Severe hemostatic defects that occur during massive transfusion are related to the volume of blood transfused, preexisting hemostatic abnormalities, concomitant pathologic changes, and therapeutic maneuvers. The relative role of each factor in the bleeding can be rapidly determined by using routine clinical laboratory tests. This determination requires an understanding of the properties of selected clotting factors, what coagulation screening tests measure, how these tests behave as the levels of factors change, and test profiles characteristic of different defects. Screening tests include platelet count, prothrombin time, partial thromboplastin time, thrombin time, and fibrinogen level. These tests are generally available on an emergent basis and can be completed within 25 minutes. The pattern associated with hemodilution is universal in massive transfusion. Patterns characteristic of the other pathologic processes that may be encountered are simply superimposed on the characteristics of hemodilution. Successful management of the contributing causes of bleeding depends on the administration of the appropriate blood components in the dose necessary to ensure that the levels of platelets and clotting factors are returned to and maintained at hemostatic levels. PMID- 10800601 TI - In-hospital mortality after acute myocardial infarction in patients with diabetes mellitus. AB - OBJECTIVES: To examine in-hospital mortality after acute myocardial infarction in patients with diabetes mellitus. METHODS: All patients in an 800-bed teaching hospital who had a discharge diagnosis of myocardial infarction, verified by creatine kinase levels at admission, between 1991 and 1993 made up the study population. All 118 such patients who died during this period made up the case group. Two control subjects (n = 236), survivors of the hospitalization, matched by sex, age, and length of hospitalization, were selected randomly for each case. Information on the presence of diabetes mellitus, medical history, and data related to myocardial infarction were obtained through retrospective chart review. RESULTS: The mean age of all subjects in the study was 76 years. Thirty three percent of the patients in the case group and 31% of the control subjects had a history of diabetes mellitus (odds ratio = 1.04; 95% CI, 0.64-1.70), indicating that diabetes mellitus was not associated with an increased risk of in hospital death. The adjusted odds ratio was 1.10 (95% CI, 0.48-2.51) in patients with non-insulin-treated diabetes mellitus and 0.80 (95% CI, 0.34-1.86) in insulin-treated patients. Multivariate analysis, with conditional logistic regression, confirmed that known prognostic factors for myocardial infarction, rather than diabetic status, are predictive of in-hospital mortality. CONCLUSIONS: Once the effects of age are accounted for, the risk of in-hospital mortality is not greater in patients with diabetes mellitus than in patients without diabetes; however, diabetes mellitus may be an important factor for long term survival. PMID- 10800602 TI - The Brugada syndrome: a case study. PMID- 10800603 TI - Reducing time in bed after percutaneous transluminal coronary angioplasty (TIBS III). AB - BACKGROUND: This study is the third in a series of investigations on the requisite length of time that patients should be restricted to bed after coronary arteriography or percutaneous transluminal coronary angioplasty using a femoral artery approach. METHODS: A prospective, experimental-control group design with randomization was used initially to compare the incidence of bleeding between patients who remained in bed for 4 hours and patients who remained in bed for 6 hours after sheath removal following percutaneous transluminal coronary angioplasty. RESULTS: Rapid changes in the healthcare environment led to nurses collecting complete data sets for the experimental group only. The experimental group (n = 51) was 73% male and 27% female; mean age was 57 years (SD = 11.4 years). Mean time in bed was 4.1 hours (SD = 0.27 hours). Most patients (98%) did not bleed from the femoral artery access site after remaining in bed for 4 hours following sheath removal. Ninety-two percent of patients required analgesics while in bed. Mean length of stay after the angioplasty was 1.4 days (SD = 0.79 days). Bleeding occurred in one subject and was related to multiple invasive procedures and an activated clotting time of greater than 200 seconds. CONCLUSIONS: Requisite time in bed after percutaneous transluminal coronary angioplasty has been reduced to 4 hours at the University of Virginia Medical Center, the same time required for patients undergoing cardiac catheterization. Discomfort after the procedure remains to be addressed. PMID- 10800604 TI - Eye care for patients receiving neuromuscular blocking agents or propofol during mechanical ventilation. AB - BACKGROUND: The presence of a corneal reflex and the ability to maintain eye closure are instrumental in protecting the cornea. Use of neuromuscular blocking agents or propofol can result in impaired eyelid closure and loss of corneal reflex, leading to corneal exposure. The cornea is then at risk for drying, infection, and scarring, which may lead to permanent visual loss. OBJECTIVES: To determine whether applying artificial tear ointment to the eyes of paralyzed or heavily sedated patients receiving mechanical ventilation decreases the prevalence of exposure keratitis more than does passive closure of the eyelid. METHODS: A prospective, randomized control trial was done. The sample was 50 patients in the intensive care unit receiving either neuromuscular blocking agents or propofol during mechanical ventilation. In each patient, artificial tear ointment was applied to one eye; passive closure of the eyelid was used for the other eye (control eye). RESULTS: Nine patients had evidence of exposure keratitis in the untreated eye, and 2 had corneal abrasions in both the treated and the control eyes. The remaining 39 patients did not have corneal abrasions in either eye. Use of the artificial tear ointment was more effective in preventing corneal exposure than was passive eyelid closure (P = .004). CONCLUSIONS: Eye care with a lubricating ointment on a regular, set schedule can effectively reduce the prevalence of corneal abrasions in patients who are either paralyzed or heavily sedated and thus can help prevent serious complications such as corneal ulceration, infection, and visual loss. PMID- 10800605 TI - The need to know: experiences of critically ill patients. AB - BACKGROUND: Critically ill patients vary in their memories of their experience in the intensive care unit. Some have little recall and need to learn about their critical illness. Others have more vivid memories of their experiences, some of which were extremely unpleasant. Patients' not knowing what was happening may have exacerbated the unpleasant experiences. OBJECTIVES: To elicit the experience of knowing for critically ill patients and to explore the differences in perceptions between patients who were intubated and those who were not intubated during the illness. METHODS: Grounded theory was used to explore the meaning of knowing and not knowing and the process by which knowing occurs. Unstructured interviews were done with 14 patients. RESULTS: Knowing had 2 phases: the need to know (1) during and (2) after the critical illness. The first phase had 3 facets: needing information, needing to be oriented, and having confusing perceptions. The second phase had 2 facets: needing information about what had happened and piecing together events. Many experiences with knowing during and after a critical illness were similar for both intubated and nonintubated patients. The main difference was the intensity of the experience in some categories. CONCLUSIONS: Critically ill patients have a strong need to know throughout and after their time in the intensive care unit. Nurses must address this need for constant reorientation to the past and present in these patients. In addition, adequate nursing staff must be available for these patients. PMID- 10800606 TI - Acquired methemoglobinemia following benzocaine anesthesia of the pharynx. PMID- 10800607 TI - Acute gastrointestinal bleeding associated with Budd-Chiari syndrome. PMID- 10800608 TI - Gender differences in acute coronary events. AB - The most frequent cause of death among women in the United States is coronary heart disease, which claims 200,000 lives a year. The prognosis with either medical or surgical therapy is worse in females than in males. The following significant gender differences have been observed and reported: (1) the rate of early death following acute myocardial infarction is greater in women, (2) the difference between sexes remains whether or not thrombolytic therapy is used, and (3) the hospital mortality rate following coronary angioplasty, atherectomy, or bypass surgery is greater in females. The reasons for these gender differences are not clearly understood. Nevertheless, awareness of the higher morbidity and mortality in women dictates the need for early detection and more aggressive therapy of the risk factors. However, diabetes mellitus and essential hypertension are 2 well-established major risk factors for coronary disease and stroke that are more prevalent in the female gender. These 2 risk factors are cumulative and require more intensive and aggressive therapy to prevent acute vascular events, and therefore early detection is mandatory. PMID- 10800609 TI - A laser scanning confocal microscopy method. Simultaneous detection of intracellular Ca2+ and apoptosis using Fluo-3 and Hoechst 33342. AB - OBJECTIVE: To develop a simple and direct method to simultaneously determine apoptotic cells from a treated population of cells and detect the changes of intracellular Ca2+ in these apoptotic cells, in particular single ones, by confocal microscopy. STUDY DESIGN: MGC-803 cells treated with As2O3 were used as the double-staining cell model with Hoechst 33342 as a DNA probe and Fluo-3AM as a Ca2+ indicator. MGC-803 cell apoptosis induced by As2O3 was first demonstrated by DNA ladder in gel electrophoresis. Based on the difference in DNA stainability with Hoechst 33342 and corresponding fluorescence intensity between live and apoptotic cells, apoptotic cells and the changes in intracellular Ca2+ were detected at the same time by confocal microscopy. No necrotic cells in the group treated with As2O3 were found by the trypan blue exclusion test. RESULTS: The results from confocal microscope detection showed that intact and apoptotic cells were successfully recognized and the changes of intracellular Ca2+ in apoptotic and intact cells were simultaneously detected in the same sample. CONCLUSION: We provided a useful method to exactly detect changes in intracellular Ca2+ in apoptotic cells, especially in single ones, by confocal microscopy and to exclude the artifact effect of necrotic and intact cells. PMID- 10800610 TI - Discrimination between dysplastic and malignant epithelium of the ampulla of vater based on quantitative image cytometric data. AB - OBJECTIVE: To assess the ability to associate histopathologic grading with objective criteria obtained by nuclear image cytometry in epithelium of the ampulla of Vater. STUDY DESIGN: Forty-one resected ampullary specimens were studied, including 8 dysplastic ampullomas together with 22 well-differentiated and 11 poorly differentiated ampullary adenocarcinomas. The nuclei were Feulgen stained and analyzed using a computer-assisted microscope, which generated 38 quantitative variables describing chromatin texture and nuclear DNA content (DNA ploidy level). These variables were explored by discriminant analysis to determine the most stable and informative variables. Univariate analysis was performed on the four most informative ones. The whole set of variables was also subjected to principal component analysis in order to characterize intragroup and intergroup heterogeneity. RESULTS: The univariate analysis defined two morphonuclear variables (related to nuclear chromatin distribution) discriminating between dysplasia and well-differentiated cancers. Aneuploidy occurrence was associated with discrimination between well-differentiated and poorly differentiated cancers. CONCLUSION: While alterations in chromatin distribution may be an early event in the malignant degeneration of this epithelium, alterations in nuclear DNA content should correspond to a later phenomenon. Quantification of these features can be exploited to assist in diagnosis. PMID- 10800611 TI - Intranuclear localization of proliferating cell nuclear antigen during the cell cycle in renal cell carcinoma. AB - OBJECTIVE: To investigate, with laser scanning cytometry (LSC), proliferating cell nuclear antigen (PCNA) expression during the cell cycle in renal cell carcinoma. STUDY DESIGN: DNA ploidy and intracellular localization of PCNA in renal cell carcinoma were determined using LSC and immunohistochemistry. The subjects were nine patients who had received surgery for renal cell carcinoma. After DNA ploidy analysis, the glass slides were restained by immunohistochemistry of PCNA. LSC allowed direct observation of PCNA localization during the cell cycle because we could obtain immunohistochemical staining of PCNA as a function of cell cycle phase for individual cells. RESULTS: PCNA was not demonstrated in the nuclei of G0/G1 cells. PCNA expression increased from the S phase of the cell cycle. PCNA rapidly degraded at the end of the G2 phase. In the late G2 and M phase, PCNA was not detected in almost any nucleus. CONCLUSION: LSC allows morphologic observation of the intracellular distribution of PCNA during the cell cycle in renal cell carcinoma. PMID- 10800612 TI - Image analysis of mesothelioma. I. differentiation of mesothelioma from adenocarcinoma of the lung. AB - OBJECTIVE: To explore the usefulness of nuclear micromorphometric analysis for the differentiation between epithelial mesothelioma and metastatic adenocarcinoma in the chest wall. STUDY DESIGN: High-resolution images of 2,100 nuclei from 27 cases of epithelial mesothelioma and 15 cases of adenocarcinoma of the lung were recorded. Stepwise discriminant analysis and a nonparametric classifier were applied to derive estimates for a case diagnosis correct classification rate. RESULTS: Nuclei from epithelial mesothelioma and adenocarcinoma of the lung showed statistically significantly different properties, but there was a region of overlap in feature space such that approximately 15-20% of cases could not be correctly classified. The lesion signatures derived from the mesothelioma cases with discriminant function scores that might result in case misclassification and the cases of adenocarcinoma of the lung spanned a similar range of degree of nuclear abnormality. However, the distribution of nuclear abnormality values for the mesothelioma cases has a mode at 0.87 SD from normal, whereas the distribution seen in lung adenocarcinoma cases had a mode at about 3.7 SD. CONCLUSION: Cases of epithelial mesothelioma and adenocarcinoma of the lung have nuclei with a wide range of deviation from normal in the spatial and statistical distribution of their nuclear chromatin. For approximately 80% of cases, correct case classification can be provided by nuclear micromorphometric analysis. Cases of epithelial mesothelioma with highly abnormal nuclei overlap in feature space with nuclei from adenocarcinoma of the lung. However, it is possible that characterization by a lesion signature may allow correct assignment for those cases. PMID- 10800613 TI - Honest assessments of automatic learning algorithm performance. AB - OBJECTIVE: To compare methods of evaluating probabilistic predictors in systems that learn from examples. STUDY DESIGN: The performance of four automatic learning algorithms, representing current machine learning technology, were assessed using four methodologies in the task of separating normal squamous intermediate cervical cells from all other segmented objects in digital images. Two of the methodologies were carefully constructed to model sources of variation associated with the choice of training and test sets. These assessments were statistically compared with assessments using both standard and a modified version of cross-validation. RESULTS: The investigation illustrates the tradeoffs involved in obtaining statistical rigor as compared with the cost of collecting data. While cross-validation makes frugal use of data, it can produce misleading assessments of algorithm performance in terms of both bias and variance. The modified version produces more reliable assessments but in some cases may also be misleading. CONCLUSION: We suggest that users of learning algorithms should exercise judicious care in evaluating learning algorithm performance in order to avoid unnecessary bias and large variance in their assessments. PMID- 10800614 TI - DNA/nuclear protein content in the evaluation of cell cycle modifications during colon carcinogenesis. AB - OBJECTIVE: To investigate the colorectal adenomacarcinoma sequence by biparametric DNA/nuclear protein flow cytometry with the aim of evaluating cell cycle modifications during carcinogenesis. STUDY DESIGN: Paraffin-embedded specimens of 27 adenomas with mild/moderate dysplasia, 20 adenomas with severe dysplasia/intramucosal adenocarcinomas, 28 adenocarcinomas and 14 normal colon mucosa specimens were analyzed by biparametric DNA/nuclear protein content flow cytometric analysis in order to evaluate cell cycle modifications during colorectal carcinogenesis. RESULTS: The mean G0-G1A fraction of the cell cycle was 50.6% (SD +/- 17.2), 25.7% (SD +/- 15.1), 27.8% (SD +/- 11.7) and 29% (SD +/- 13.8) for normal mucosa, adenomas with mild/moderate dysplasia, adenomas with severe dysplasia and adenocarcinomas, respectively. The difference between normal mucosa and the other groups was statistically significant (P < .05), while no significant differences were detectable between adenomas with different degrees of dysplasia and adenocarcinomas. CONCLUSION: Our results show a decrease in G0 G1A in adenomas with mild/moderate dysplasia, suggesting that modification of the cell cycle may represent an early step in colon carcinogenesis, and they support the hypothesis that disregulation of cell cycle-controlling genes is an early event in the adenoma-carcinoma sequence. PMID- 10800615 TI - Quantitative cytology in ovarian carcinoma ascitic fluids. AB - OBJECTIVE: To assess the value of quantitative methods in the differential diagnosis between ovarian carcinoma cells and mesothelial cells in ascitic fluids. STUDY DESIGN: Ninety ascitic fluid samples, previously reported as positive for ovarian carcinoma (30 cases), suspicious for malignancy (30) and negative for malignancy, containing only reactive mesothelial cells (30), were retrieved from the files. In each of these specimens the nuclear area, perimeter, roundness and shape coefficient of 100 cells were determined at 630 x magnification. Statistical analysis was performed using analysis of variance and, for multiple comparisons, the Student-Newman-Keuls technique. RESULTS: Mean values for nuclear area and perimeter were higher in malignant cells as compared to reactive mesothelial cells, whereas those for roundness and shape coefficients were lower. All differences were statistically significant, the former two at a .05 level and the latter at the .001 level. CONCLUSION: Quantitative methods can reliably support the differential diagnosis between ovarian carcinoma cells and mesothelial cells in ascitic fluid specimens. PMID- 10800616 TI - Differential cell analysis of cytocentrifuged bronchoalveolar fluid samples affected by the area counted. AB - OBJECTIVE: To investigate variations in the differential cell counts between the quadrants of cytocentrifuged bronchoalveolar lavage (BAL) fluid preparations and to evaluate the diagnostic impact of these differences in interstitial lung diseases (ILD). STUDY DESIGN: BAL fluid samples obtained from 30 patients suspected of having ILD or pneumonia were cytocentrifuged and additionally stained with May-Grunwald-Giemsa stain. Two observers differentiated 200 cells in each quadrant as well as in a circular pattern around the center of the cytocentrifuge spot. RESULTS: Lymphocytes and alveolar macrophages were not randomly distributed on the cytocentrifuge spot. Ten samples of patients with histologically confirmed ILD were selected to test the diagnostic impact using a validated computer program. The predicted diagnosis did not correspond to the histologic diagnosis for one quadrant from 1 of these 10 samples (sarcoidosis instead of idiopathic pulmonary fibrosis), whereas the differential cell counts performed around the center of the cytocentrifuge spot provided the correct diagnosis in all cases. CONCLUSION: BAL fluid differential cell counts varied between the quadrants of the cytocentrifuge spot. The center of the cytocentrifuge spot appeared to be the most reliable area. Therefore, cell counting is recommended in a circular pattern around the center of the cytocentrifuge spot. PMID- 10800617 TI - Effect of melanin bleach on Feulgen-DNA microdensitometry in pigmented lesions. AB - OBJECTIVE: To investigate the effect of melanin bleach on Feulgen-DNA microdensitometry in pigmented melanocytic lesions. STUDY DESIGN: Twenty banal compound nevi with various grades of pigmentation were bleached by 0.5% and 1% KMnO4 for 0 to 20 minutes and by 10% H2O2 for 24 hours prior to Feulgen staining. DNA microdensitometry was performed by video image analysis to measure the integrated optical density (IOD) in nuclei from nevomelanocytes, lymphocytes and spinous keratinocytes. The DNA index of nevomelanocytes was calculated using spinous keratinocytes as the diploid controls. RESULTS: There were significant decreases in IOD (P < .05) in the nuclei of nevomelanocytes, lymphocytes and spinous keratinocytes after treatment with 1% KMnO4 for 5 and 10 minutes, but no significant changes were detected after treatment with 0.5% KMnO4 for 5 and 10 minutes. Severe tissue damage was observed in the Feulgen-stained slides treated with 1% KMnO4 for 15 and 20 minutes and with 10% H2O2 for 24 hours. There was no significant change in DNA index in any bleached sets measured. CONCLUSION: KMnO4 can affect Feulgen-DNA content if used in high concentrations or for long periods of incubation. The DNA index, which is derived from internal controls, is not affected by the bleach procedure. PMID- 10800618 TI - Topologic distribution of different types of neurons in the human putamen. AB - OBJECTIVE: To test the assumption that the various types of neuron in the human putamen appear to be randomly distributed and to quantify the way in which they are arranged, stochastic geometry, multivariate analysis and the interactive evaluation technique were employed. STUDY DESIGN: Twenty-seven human putamina without demonstrable signs of neurologic change were dissected out, fixed in 4% formalin and embedded in paraffin. The 20-micron paraffin sections were stained in an aldehyde-fuchsin and cresyl-violet solution, which makes it possible to distinguish between seven different neuron populations in the putamen. The gravity centers, size and form factors of these neurons were determined morphometrically under a light microscope. The data obtained were used to calculate the spatial distribution of the neurons by interactive and structure analytical methods. RESULTS: Visual point field analysis revealed an irregular arrangement of the different types of neurons. Point process analysis detected a significant hard core process of type 1 and a cluster process of type 6 neurons. With nearest neighborhood analysis, significant differences were found between certain populations of neurons and Poisson processes. Comparison of the results of multivariate cluster analysis with the investigator-dependent results of visual point field analysis showed clear differences. CONCLUSION: By means of structure analytical methods, the arrangement of different populations of neurons can be demonstrated. Some neuronal distributions are detectable only by using one of these techniques. The question of random or nonrandom distribution of the neurons in the human putamen can now be answered definitively: arrangement of the different populations of neurons is structured. PMID- 10800619 TI - Factor analysis of confocal image sequences of human papillomavirus DNA revealed with fast red in cervical tissue sections stained with TOTO-iodide. AB - OBJECTIVE: To visualize and localize specific DNA sequences by fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM) and factor analysis of biomedical image sequences (FAMIS). STUDY DESIGN: Human papillomavirus (HPV) DNA was identified in cervical tissue sections with biotinylated DNA probes recognizing the whole genome of HPV DNA types 18 and 16, and DNA-DNA hybrids were revealed by streptavidin-alkaline phosphatase and Fast Red (FR). Cell nuclei were counterstained with TOTO-iodide. Image sequences were obtained using successive dynamic or spectral sequences of images on different optical slices from CLSM. The location of fluorescent signals inside tissue preparations was determined by FAMIS and/or selection of filters at emission. Image sequences were summarized into a reduced number of images, called "factor images," and curves, called "factors." Factors estimate spectral patterns and depth emission profiles. Factor images correspond to spatial distributions of the different factors. RESULTS: We distinguished between FR and nucleus staining in HPV DNA hybridization signals by taking into account differences in their spectral patterns and improved visualization by taking into account differences in their focus (depth emission profiles). CONCLUSION: FAMIS, together with CLSM, made possible the detection and characterization of HPV DNA sequences in cells of cervical tissue sections. PMID- 10800620 TI - Flow cytometric DNA analyses of epithelial dysplasia of the esophagus. AB - OBJECTIVE: To investigate, with flow cytometry, DNA aneuploidy as a marker of early carcinogenesis in dysplastic esophageal lesions. STUDY DESIGN: DNA content of exfoliated cells from 789 cases of esophageal dysplasia (including mild dysplasia, 195 cases; moderate dysplasia, 383 cases; and severe dysplasia, 211 cases) was determined with a FACS 420 flow cytometer. RESULTS: Cellular DNA content was closely related to the severity of dysplasia. The carcinogenesis rate in patients with dysplasia showed that DNA aneuploidy was significantly higher than in patients showing DNA diploidy. CONCLUSION: DNA aneuploidy in dysplastic lesions is a very important early signal of carcinogenesis. Patients with dysplastic lesions showing DNA aneuploidy should be treated and closely followed. PMID- 10800621 TI - 3-D Golgi and image analysis of the olfactory tubercle in schizophrenia. AB - OBJECTIVE: To examine morphologic changes in the olfactory tubercle (OT) spiny neurons and astrocytes in schizophrenia (Sch) by means of quantitative 3-D Golgi and immunocytochemical studies. STUDY DESIGN: Free-floating vibrotome sections of postmortem brain tissue from 10 controls and 12 Sch cases were used for Golgi study and glial fibrillary acidic protein (GFAP) immunocytochemistry. A gray level image analysis was applied for quantitative estimation of GFAP-positive astrocytes on uniform, randomly sampled sections. This method is effective for low-contrast objects on an uneven background. Golgi-impregnated OT spiny neurons were analyzed both qualitatively and quantitatively in three dimensions with a semiautomated microscope-computer system. From digitized image of the neurons, various metric parameters were estimated to characterize the dendritic tree. RESULTS: In cases of Sch, degenerative changes in the dendrites of OT spiny neurons were revealed. A decrease in the maximal radius of the dendritic tree and total length of dendrites were accompanied by an increase in the length density of dendrites. Hypertrophy and a more-intensive GFAP reaction of astrocytes were found in OT of Sch. CONCLUSION: Based on these results, one can hypothesize that OT spiny neurons in Sch are involved in the process of dendritic reorganization, including degenerative changes in dendrites. PMID- 10800622 TI - Morphometric analysis of AgNORs in uveal malignant melanoma. PMID- 10800623 TI - Syntactic comprehension, verbal memory, and calculation abilities in Spanish English bilinguals. AB - This article analyzes the interfering effect of the second language (L2) on the first language (L1) in native Spanish speakers living in the United States. We examined 3 linguistic aspects: (a) syntactic comprehension, (b) verbal memory, and (c) calculation abilities. We carried out 2 different studies. In the 1st study, we studied syntactic understanding in 50 Spanish-English bilinguals. For all participants, L1 was Spanish and L2 was English, and all learned English early in life and had attended English schools. Results for the Spanish Syntactic Comprehension Test (Marcos & Ostrosky, 1995) were compared with the normative results obtained with 40 Spanish monolingual participants. We observed that the closer to the English syntax the sentences were, the easier it was for the participants to understand them. Participants who had been exposed to English between the ages of 5 and 12 outperformed participants exposed to English before 5 years of age. Language preference correlated with syntactic comprehension. Women outperformed men. In the 2nd study, verbal memory and calculation abilities were examined in L1 and L2 in a group of 85 Spanish-English bilinguals. Parallel versions of the different tests were administered in Spanish and English. The results indicated some significant differences between the 2 languages in several verbal learning and calculation ability subtests. Most of the verbal memory subtests were better performed in L1. Scores on tasks measuring speed and calculation accuracy were higher in the participant's native language. Best spoken language proved to be a significant variable in some verbal memory subtests performed in English but not in Spanish. We analyze implications of bilingualism in neuropsychological testing. We also present some suggestions to minimize the bilingualism effect. PMID- 10800624 TI - Verbal fluency and repetition skills in healthy older Spanish-English bilinguals. AB - The influence of bilingualism on cognitive test performance in older adults has received limited attention in the neuropsychology literature. The aim of this study was to examine the impact of bilingualism on verbal fluency and repetition tests in older Hispanic bilinguals. Eighty-two right-handed participants (28 men and 54 women) with a mean age of 61.76 years (SD = 9.30; range = 50-84) and a mean educational level of 14.8 years (SD = 3.6; range 2-23) were selected. Forty five of the participants were English monolinguals, 18 were Spanish monolinguals, and 19 were Spanish-English bilinguals. Verbal fluency was tested by electing a verbal description of a picture and by asking participants to generate words within phonemic and semantic categories. Repetition was tested using a sentence repetition test. The bilinguals' test scores were compared to English monolinguals' and Spanish monolinguals' test scores. Results demonstrated equal performance of bilingual and monolingual participants in all tests except that of semantic verbal fluency. Bilinguals who learned English before age 12 performed significantly better on the English repetition test and produced a higher number of words in the description of a picture than the bilinguals who learned English after age 12. Variables such as task demands, language interference, linguistic mode, and level of bilingualism are addressed in the Discussion section. PMID- 10800625 TI - Sensitivity and specificity of the Mini-Mental State Examination in a Spanish speaking population. AB - The Mini-Mental State Examination (MMSE; Folstein, Folstein, & McHugh, 1975) was given to a total of 430 normal participants divided into 3 age ranges (16-50, 51 65, and 66-89) and 4 educational ranges (0, 1-4, 5-9, and > or = 10 years). The educational level effect was notoriously stronger than the age effect. Normal illiterate participants obtained scores that would correspond to severe cognitive alterations (M = 17.67); low education participants (1-4 years) would be classified with moderate cognitive alterations (M = 20.61). Sensitivity and specificity of the MMSE were established. Low sensitivity and specificity were found for both the participants with 0 and 1 to 4 years of schooling, 50% and 72.73%, respectively. In participants with more than 5 years of schooling, the specificity (86.36%) and sensitivity (86.36%) indexes were higher. We concluded that the MMSE is an instrument with little diagnostic utility among participants with a low level of education. PMID- 10800626 TI - Factor analysis of the Neuropsychological Screening Battery for Hispanics (NeSBHIS). AB - The Neuropsychological Screening Battery for Hispanics (NeSBHIS; Ponton et al., 1996) was developed to provide clinicians and researchers with appropriate tests and norms to assess the Latino population. However, the construct validity of the NeSBHIS is unclear, and its clinical utility is untested. This study provides data on the construct validity of this battery via factor analytic methodology. Five factors were identified: (a) a language factor, which included the Controlled Oral Word Association Test, the Ponton-Satz Boston Naming Test, and the Escala de Inteligencia Wechsler para Adultos (EIWA; Wechsler, Green, & Martinez, 1968) Digit Span; (b) a verbal learning factor, which included all the World Health Organization--University of California, Los Angeles Auditory Verbal Learning Test scores (Trials V, VII, and VIII); (c) an attentional-mental control factor, which included the EIWA Digit Symbol, Color Trails 1 and 2, and the EIWA Block Design; (d) a visuospatial factor, made up of the Rey-Osterrieth Complex Figure Test Copy and Memory and the Raven's Standard Progressive Matrices total score; and (e) a psychomotor factor, the Pin Test. This was a stable factor structure, suggesting that the NeSBHIS has a robust construct validity. PMID- 10800627 TI - Neurobehavioral assessment of attention deficit hyperactivity disorder in a Colombian sample. AB - We present a factor analytic study of the Conners Rating Scales for parents and teachers in this article. A comparison is established with the original factor analytic studies (Conners, 1979a, 1979b) and the results obtained by Farre-Riba and Narbona (1997), using a Spanish sample. Five hundred and forty children, ages 4 to 17, were randomly selected in Manizales, Colombia. The shortened Spanish version of the Conners Parents Rating Scale (CPRS; Conners, 1979a) and the Conners Teacher Rating Scale (CTRS; Conners, 1979b) were used. Parsimonious factor analyses for both scales were developed. Three stable factors were found in the CPRS: hyperactivity, somatic symptoms, and inattention. Twenty-two out of 48 items were significantly saturated by these 3 factors. In the CTRS, 4 different factors were found that accounted for 63.5% of the total variance: uncontrolled temperament, inattention, hyperactivity, and difficulties in social relationships. Twenty out of 28 items were significantly saturated by these 3 factors. Factor structure was closer to the Spanish Farre-Riba and Narbona report than to the original findings. We concluded that the CPRS and the CTRS Spanish versions, when used by Colombian children and adolescents, do not seem to evaluate exactly the same underlying behavioral dimensions. We propose selecting only 22 items of the CPRS and 20 items of the CTRS (brief versions) for further epidemiological and clinical use. PMID- 10800628 TI - Coherence in short narratives written by Spanish-speaking children with reading disabilities. AB - A novel analysis of coherence using a combination of three criteria (syntactic connexity, pragmatic complexity, and rhetorical well-roundedness) was applied to short narratives produced by a group of 60 Spanish-speaking children of different ages and grades with reading disabilities and compared to those produced by normal children. We posit a scale of 6 degrees of increasing coherence. This feature of children's writing, together with 2 others (viz. number of propositions, or "story points," recovered and number of words employed), was compared to features of children's reading by means of discriminant analysis in relation to age. We show that the combination of age, words read per minute, and degree of coherence achieved an optimal discrimination of the 2 groups. PMID- 10800629 TI - Pharmacological effects of triazolo[4,3-d] and tetrazolo[1,5 a][1,4]benzodiazepines on the central nervous system. AB - A series of tricyclic 1,4-benzodiazepines, fundamentally 5H-s-triazolo[4,3-d] and tetrazolo[1,5-a][1,4]benzodiazepines, were tested for acute toxicity and central nervous system effects in mice. All of these compounds, but especially 3, 4 and 5, showed a clear central depressant activity. Compounds 3, 4 and 5 also showed a moderate activity in antagonizing the deleterious effects produced by cardiazol but were ineffective in counteracting those elicited by strychnine. These same compounds 3, 4 and 5 presented a remarkable benzodiazepine-like action with a pharmacological profile similar to that of chlordiazepoxide. PMID- 10800630 TI - Bioavailability investigation of two different oral formulations of Tetrazepam. AB - Two different oral tetrazepam (CAS 10379-14-3) formulations (Tetrazepam ratiopharm film-coated tablets as test preparation and tablets of a reference preparation marketed in France) were investigated in 20 healthy volunteers in order to prove bioequivalence between these preparations. A single 50 mg oral dose was given according to a randomised two-way crossover design in the fasted state. Blood samples for determination of tetrazepam plasma concentrations were collected at pre-defined time points up to 96 h following drug administration. A washout period of 14 days separated both treatment periods. Tetrazepam plasma concentrations were determined by means of a validated LC-MS/MS method. Values of 3873.08 ngh/ml (test preparation) and 3930.69 ngh/ml (reference preparation) for the parameter AUC0-infinity demonstrate an nearly identical extent of drug absorption. Maximum concentrations (Cmax) of 482.08 ng/ml and 465.14 ng/ml were achieved for test and reference preparation. Time to reach maximum plasma concentration (tmax) was 1.39 hours for both preparations. Cmax and AUC0-infinity values were tested parametrically by an analysis of variance (ANOVA). Bioequivalence was concluded if the 90% confidence intervals of the T/R-ratios were in the range of 80%-125% for AUC0-infinity and 70%-143% for Cmax. Based on the results obtained in this study, bioequivalence between the test and the reference preparation was demonstrated. PMID- 10800631 TI - Cardiovascular pharmacological studies on JTV-506, a new potassium channel opener. 2nd communication: effects on blood pressure in conscious animals. AB - The effects of JTV-506 ((-)-(3S,4R)-2,2-bis(methoxymethyl)-4-[(1,6-dihydro-1 methyl-6-oxo-3- pyridazinyl)amino]-3-hydroxychroman-6-carbonitrile hemihydrate, CAS 170148-29-5), a novel coronary vasodilator, on blood pressure were evaluated in conscious dogs and rats. In conscious dogs, JTV-506 (0.01-1 mg/kg p.o.), levcromakalim (0.01-1 mg/kg p.o.) and nifedipine (3-30 mg/kg p.o.) elicited an increase in double product, whereas nicorandil (1-10 mg/kg p.o.) did not affect the double product. The JTV-506-induced increase in double product was abolished by a beta-blocker, propranolol, suggesting that this increase in double product may be due to augumentation of heart rate by sympathetic nerves which mediate the baroreflex. The doses at which JTV-506 increased coronary blood flow in a previous study were lower than the doses required to increase the double product. JTV-506 did not have a crucial influence on electrocardiogram. In conscious rats, orally administered JTV-506, levcromakalim, nicorandil and nifedipine reduced blood pressure and increased heart rate dose dependently. These effects were more remarkable in hypertensive rats than in normotensive rats. JTV-506, a new potassium channel opener, seems to be relatively free of any hemodynamic effects. PMID- 10800632 TI - Effect of nicorandil upon different guinea-pig and rat isolated organ preparations in vitro. AB - A study of the effect of nicorandil (N-2-(hydroxyethyl)nicotinamide nitrate, CAS 65141-46-0), a potassium channel and guanylatecyclase activator, upon preparations of rat was deferens and uterus, and guinea pig ileum was performed. Nicorandil does not modify rat isolated was deferens responses to noradrenaline (norepinephrine) and potassium. The drug exerts a non-competitive antagonist effect upon rat isolated uterus response to serotonin, histamine, oxytocin, and, at high concentrations, inhibits guinea-pig isolated ileum responses to acetylcholine, histamine, 4-aminopyridine and potassium. PMID- 10800633 TI - Preclinical pharmacology of desloratadine, a selective and nonsedating histamine H1 receptor antagonist. 1st communication: receptor selectivity, antihistaminic activity, and antiallergenic effects. AB - Desloratadine (descarboethoxyloratadine, CAS 100643-71-8) is an active metabolite of loratadine (CAS 79794-75-5) that exhibits qualitatively similar pharmacodynamic activity with a relative oral potency in animals 2.5-4 times greater than loratadine. Its antihistaminic effect lasts 24 h. Desloratadine was shown to be a selective H1 antagonist with more potent antihistaminic activity in vitro than either loratadine or terfenadine (CAS 50679-08-8), as indicated by its displacement of 3H-mepyramine from H1 receptors in rat brain, guinea pig brain, and guinea pig lung, and by its antagonism of histamine-induced contractions of guinea pig ileum. Antihistaminic activity and anitallergic effects also were observed in vivo. After oral administration, desloratadine was 2.5 to 4 times more potent than loratadine in protecting against histamine-induced lethality in the guinea pig and paw edema in the mouse; after topical administration, it was almost 10 times more potent in antagonizing histamine-induced increases in nasal microvascular permeability in the guinea pig. Histamine-induced changes in pulmonary resistance and compliance were also prevented by oral administration of desloratadine and loratadine in the monkey. An oral antiallergic effect was demonstrated by important reductions of acute bronchospasm in the allergic monkey and potent inhibition of allergic cough in the guinea pig. These preclinical studies provide evidence that desloratadine is an antihistaminic agent with a greater potency than loratadine and, together with results from numerous published studies, suggest an antiallergic effect of desloratadine. PMID- 10800634 TI - Synthesis and analgesic effects of 5-[4-(arylpiperazin-1-yl)alkylamino]-4-benzyl 3-methyl-1,2-oxa zin-6-one s. AB - A series of 5-[4-(arylpiperazin-1-yl)alkylamino]-4-benzyl-3-methyl-1,2-oxaz in-6 ones was synthesized and evaluated for analgesic activity. The structures of these new oxazine derivatives were confirmed by IR, 1H-NMR spectra and by elemental analysis. The three most active compounds, 3c, 3e and 3g possessed significant antinociceptive effects in the phenylbenzoquinone-induced wrigthing test (PBQ-test) in mice, with ED50 values ranging from 19.7 to 68.0 mg/kg i.p. In addition these compounds presented a low toxicity (LD50 > 800 mg/kg i.p.) and did not significantly reduce the spontaneous locomotor activity of mice. They interacted in a synergistic manner with morphine but nevertheless each compound presented its own profile. Thus the analgesic activity of 3c and 3e was naloxone sensitive, suggesting in mu opioidergic mechanism. Otherwise 3c and 3d analgesia was attenuated by oral administration of yohimbine and therefore seemed to be mediated via noradrenergic pathway. Finally, 5-hydroxytryptophan associated to carbidopa only potentiated 3e analgesia, demonstrating an involvement of a serotoninergic mechanism. PMID- 10800635 TI - Biochemical model reactions indicative of inflammatory processes. Activities of extracts from Fraxinus excelsior and Populus tremula. AB - All processes of oxygen activation include very reactive intermediates. Therefore, aerobic cells must cope with--and to some extent also adapt to- oxidative stress provoked for example by infections or intoxications, where these reactive intermediates accumulate. All inflammatory processes include such oxygen activating processes where reactive oxygen species (ROS) are produced. Dependent on the strength of these impact(s), several symptoms indicate the deviation from normal, steady-state metabolism. Intrinsic radical scavenging processes or compounds administered with food thus have to warrant metabolic control within certain limits. Antioxidants which in many cases are free radical scavengers or quenchers of activated states comprise a vast number of classes of organic molecules including phenolics as the most prominent ones. In this publication the activities of extracts from Fraxinus excelsior, Populus tremula and Solidago virgaurea as components of the drug Phytodolor and their mechanisms of protection from oxidative damage are summarized. In addition, new results on tyrosine nitration, a process characteristic for sites of inflammation, and its inhibition by these plant extracts, is reported. PMID- 10800637 TI - Hypocholesterolaemic and antiatherosclerotic effects of tetra-iso-propyl 2-(3,5 di-tert-butyl-4-hydroxyphenyl)ethyl-1,1-diphosphonate (SR-9223i). AB - Tetra-iso-propyl 2-(3,5-di-tert-butyl-4-hydroxyphenyl)ethyl-1,1-diphosphonate (CAS 126411-13-0, SR-9223i) is a member of a new class of compounds with multiple antiatherosclerotic activities. This report not only describes the cholesterol lowering properties in four species of animals fed normocholesterolaemic diets but also reductions in lipid deposition in the arteries of cholesterol-fed New Zealand white rabbits following the administration of SR-9223i. Plasma cholesterol concentrations were reduced in mice by 27% (200 mg/kg/day administered in the diet for 10 days), in hamsters by 33% (200 mg/kg/day administered in the diet for 8 days), in dogs by 16% (25 mg/kg/day p.o. for 28 days) and 23% (75 mg/kg/day p.o. for 28 days) and in monkeys by 22% (25 mg/kg/day p.o. for 28 days). Further, the deposition of cholesterol, especially in the esterified form, in the aortae of cholesterol-fed New Zealand white rabbits was inhibited by SR-9223i (50 and 100 mg/kg/day p.o.). At the higher dose, the cholesteryl ester content of the aorta was half that of control animals. SR 9223i, at both doses, also inhibited the accumulation of cholesterol in the liver. SR-9223i has been shown to suppress HMG CoA reductase activity, inhibit ACAT activity and prevent lipid oxidation. These activities, demonstrated in vitro, have now been shown to translate into lipid lowering and antiatherosclerotic activities in vivo. PMID- 10800636 TI - Antiapoptotic effects of propolis extract and propol on human macrophages exposed to minimally modified low density lipoprotein. AB - An aqueous extract of propolis and the phenolic component of propolis, propol, were assayed for antioxidative and antiapoptotic properties. Both additions inhibited Cu(2+)-initiated low density lipoprotein (LDL) oxidation as characterized by a reduction of the lag time, reduced the increase of relative electrophoretic mobility during oxidation and markedly diminished apoptosis of human macrophages exposed to minimally modified (mmLDL). Moreover, aqueous propolis extract and propol blocked the mmLDL-induced decrease of glutathione (GSH) and the activation of the transcription factor NF-kappa B in these cells. The potent phenolic antioxidant propol thus expands the capability of cells to neutralize oxidative stress and to prevent apoptosis and is therefore suggested to significantly contribute to the antiinflammatory and antioxidative effects of propolis. PMID- 10800638 TI - Effect of meropenem on the vascular permeability factor produced by Acinetobacter baumannii. AB - Eleven Acinetobacter baumannii strains produced a toxic substance--the vascular permeability factor--in the culture medium. Intradermal injection of this substance enhanced vascular permeability in the rabbit skin. The extent of permeability reactions varied from 0.28 cm2 to 1.61 cm2. Changes in the permeability factor activity of four A. baumannii strains after treatment with meropenem (CAS 96036-03-2) at suprainhibitory concentrations (2x, 4x or 8x) minimum inhibitory concentration (MIC) or supra-subinhibitory ones (2-8x MIC + 0.2x MIC) were tested in vitro. Meropenem at all suprainhibitory concentrations (with the exception of 8x MIC for one strain) was almost ineffective. Alterations in this activity were in the range of 93% to 106% of the control values. Supra subinhibitory concentrations of meropenem significantly increased the permeability factor activity (to 150%-176% of the control values). These findings indicate that meropenem mainly at supra-subinhibitory concentrations can in vitro interfere with the vascular permeability factor produced by A. baumannii. PMID- 10800639 TI - Effect of a combination of the new antischistosomal drug Ro 15-5458 and praziquantel on different strains of Schistosoma mansoni infected mice. AB - The possible additive or synergistic effects of both praziquantel (CAS 55268-74 1) and a new antischistosomal drug, Ro 15-5458 (10-(2-diethylamino)thyl)-9 acridanone(thiazolidin-2-yl-i dene)hydrazone, CAS 92928-47-7) were studied in two different strains of Schistosoma mansoni infected mice, namely CD susceptible and SO4 resistant strains. Assessment of cure was performed using the following parameters: hepatic and intestinal tissue egg load and distribution, oogram changes in the small intestine and histopathological examination of the mice livers. In this study, a combination was used between 1/3 the curative doses of praziquantel and Ro 15-5458. This combination therapy proved to be beneficial as regards the percentage parasite reduction and hepatic worm shift (99.4% and 100%, respectively, in the CD susceptible mouse strains, compared to 84.1% and 34.8% in the SO4 resistant strains). Treatment with subcurative doses of praziquantel and Ro 15-5458 resulted in 78.6% intestinal dead ova and 21.4% mature ones. This score shifted to 98.6% and 1.4% dead and mature ova, respectively, in the SO4 resistant strains. Again the range of liver granulomata in the CD susceptible and SO4 resistant strains receiving subcurative doses of both drugs was 4-6 and 2-5, respectively, in five successive low power fields, while in the infected untreated control mice, this range reached 8-11 and 5-9, respectively. Histopathological sections of the liver revealed a small fibrocellular granuloma with few inflammatory cells and excess fibrous collagen tissue deposition in animals undergoing the combination therapy. This contrasts with the large fibrocellular granulomata seen in the infected untreated control mice. These results may be of value in endemic areas of schistosomiasis, due to the unexpected emergence of drug resistance against the currently used antischistosomal drug, praziquantel in these areas. PMID- 10800641 TI - [Paul Ehrlich's methods for deciphering the immune defence. One method leads only to an incomplete understanding]. AB - Paul Ehrlich's routes to decipher the immune defence came on the one side as a black box consideration from differences in the potency of the final result and on the other side from connections between the single events. In consequence of the assumption of linear interactions between the single events it was believed, as in the case of pharmacology, that Ehrlich's black box consideration from the final result could be dispensed with. The single event would be already representative for the proceedings in general. But the assumption is wrong and Ehrlich's black box consideration from the final result has to be applied again to decipher the real causes of sufficient or insufficient immune defences, of immune diseases, of allergies etc. We have to learn again, how to use two contrary considerations, which likewise have their rights, to understand the immune defence. PMID- 10800640 TI - General pharmacology of the new antiviral agent SK 1899. AB - The general pharmacological properties of 2-amino-9-(3-acetoxymethyl-4 isopropoxycarbonyloxybut-1-yl)purine (CAS 247081-81-8, SK 1899), a new potential antiviral agent, were investigated in mice, rats, guinea pigs, rabbits, and dogs. The oral administration of 50, 150, and 500 mg/kg of SK 1899 had no effects on the central nervous system except that it slightly increased the spontaneous locomotor activity in mice at a dose of 500 mg/kg. SK 1899 did not disturb either the spontaneous motility or contractor-induced contraction of the isolated organs such as guinea pig ileum, rat uterus, guinea pig vas deferens, and guinea pig trachea at concentrations up to 10(-4) mol/l. It slightly increased the contractile force in the isolated guinea pig atrium at a concentration of 10(-4) mol/l. Following intravenous infusion of 5, 15, and 50 mg/kg of SK 1899 to anesthetized dogs, it did not change the mean arterial pressure, heart rate, left ventricular systolic pressure (LVSP), and respiratory rate, while it slightly increased the left ventricular positive dP/dtmax (LV + dP/dtmax) at a dose of 50 mg/kg. SK 1899 did not induce any significant changes in the intestinal charcoal meal transit in mice, basal gastric juice secretion in rats, and renal function in rats. It did not affect the blood coagulation system and phenolsulfonphthalein secretion in rats. These findings suggest that SK 1899 has a very low potential to induce any adverse pharmacological effects at the doses showing antiviral activity. PMID- 10800642 TI - Allogeneic peripheral blood stem cell transplantation for hematologic diseases. PMID- 10800643 TI - Unrelated donor marrow transplantation for treatment of childhood hematologic malignancies-effect of HLA disparity and cell dose. PMID- 10800644 TI - Haploidentical transplantation. AB - When allogeneic transplant is indicated and a MSD is not available, most centers first seek an unrelated donor. However, one must avoid undue delays when a donor is not available and family members should be typed to identify the preferred donor amongst the family. Since a family donor can provide access to transplant for almost every patient, a greater effort should be encouraged to improve and expand haploidentical transplantation. The advantages, outlined in Table 1, suggest compelling reason to perfect the use of haploidentical related donors. PMID- 10800645 TI - Umbilical cord blood hematopoietic stem cell transplantation. Eurocord-Cord Blood Transplant Group. PMID- 10800646 TI - Non myeloablative "mini transplants". AB - The efficacy of graft-vs.-leukemia induction to treat relapses after allogeneic progenitor cell transplant in a variety of hematologic malignancies suggest that it may be possible to use the graft versus leukemia as primary therapy for these malignancies without the need of myeloablative therapy. This type of strategy should be explored initially in patients considered ineligible for conventional myeloablative therapies either because of age or concurrent medical conditions. GVHD remains a major obstacle that needs to be overcome. Although a potentially lower level of inflammatory cytokines may be present after non-myeloablative therapies, fatal GVHD still occurs. Methods to diminish GVHD after allogeneic transplant include selective T-cell depletion (39-43) and transduction of donor T cells with Herpes simplex virus thymidine kinase which renders these cells sensitive to ganciclovir treatment (see chapter 16). We and others have demonstrated that nonablative chemotherapy using fludarabine combinations is sufficiently immunosuppressive to allow engraftment of allogeneic blood progenitor cells. Patients could then receive graded doses of donor lymphocytes without rejection, to mediate GVL. Ideally, this therapy could be titrated to levels of residual malignant cells using sensitive detection techniques. This novel approach to therapy would reduce the toxicity of the transplant procedure, allow it to be administered more safely to debilitated patients and possibly extend the use of transplantation to older patients who are not presently eligible for BMT procedures. Other possible indications include treatment of non malignant disorders and induction of tolerance for solid organ transplantation. PMID- 10800647 TI - Allogeneic hematopoietic stem cell transplantation in recipients of cellular or solid organ allografts. PMID- 10800648 TI - Allogeneic peripheral blood progenitor cell transplantation in solid tumors. PMID- 10800649 TI - Hematopoietic stem cell transplantation of multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus. PMID- 10800650 TI - Advances in the control of cytomegalovirus disease in bone marrow transplant patients. PMID- 10800651 TI - Adoptive immunotherapy for EBV-associated malignancies. PMID- 10800652 TI - Adoptive immunotherapy using donor leukocyte infusions to treat relapsed hematologic malignancies after allogeneic bone marrow transplantation. PMID- 10800653 TI - Clinical use of irradiated donor lymphocytes in bone marrow transplantation. PMID- 10800654 TI - Dendritic cells and their clinical applications. PMID- 10800655 TI - Engineering hematopoietic grafts using elutriation and positive cell selection to reduce GVHD. AB - A systematic approach to hematopoietic graft manipulation has minimized several of the variables inherent to allogeneic BMT. Through this approach, we have been able to significantly impact on morbidity and quality of life following allogeneic transplantation. Acute and chronic GVHD, blood product and antibiotic usage, in patient hospitalization, acuity, costs and survival (especially in patients older than 40) have been improved. The HLA barrier still presents a formidable obstacle to achieving a more widespread use of this therapy. The complications encountered in HLA matched/TCD grafts occur with even greater magnitude in the HLA-mismatched or unrelated donor setting. Several centers are now engaged in studies using TCD grafts that are augmented with high doses of CD34+ cells to ensure engraftment while reducing the incidence of GVHD (50-53). Mobilized allogeneic PBSC appear to be an excellent source of stem cells for BMT (5,6). The earlier reports showed decreased rates of GVHD, despite having T cell burdens 10 times higher than those found in unmanipulated bone marrow. However, several of these centers now report an unacceptably high incidence of chronic GVHD (along with its attendant morbidity) following allogeneic PBSC transplantation (54-55). Initial results of TCD in these PBSC grafts using CD34+ selection are disappointing in that recipients developed unexpectedly high incidences of both acute and chronic GVHD (56). No doubt, significant differences exist between marrow and PBSC ancillary cell populations. For example, two laboratories now report the presence of natural suppressor cells in these allogeneic PBSC products in both mice (57) and humans (58). Thus, the same, step wise approach would be expected to improve graft performance when using PBSC, cord blood, fetal tissue, xenografts or genetically engineered products as a stem cell source. Indeed, there are new reports of improved clinical outcome (especially in the incidence of GVHD) in the PMRD setting using both CD34+ selected (59) and sequential CD34+/CD2+ selected (60) PBSC grafts. It is hoped that future graft engineering approaches will be as successful as previous studies and will extend this form of therapy to an even larger patient population. PMID- 10800656 TI - Monoclonal antibody and receptor antagonist therapy for GVHD. PMID- 10800657 TI - Adoptive immunotherapy for leukemia: donor lymphocytes transduced with the herpes simplex thymidine kinase gene. AB - The overall goal of adoptive immunotherapy with genetically modified lymphocytes is to decrease the morbidity and mortality associated with allogeneic bone marrow transplantation. The initial data reviewed here suggest that the behavior of the allogeneic HStk transgenic cells can be modified after administration to patients. Further study is needed to identify the response rates and risks associated with this procedure. In particular, larger studies will be needed with appropriate randomization to determine if the response rate to genetically modified cells is equivalent to the response rates with unmodified cells. Wider application of these techniques in the initial setting of allogeneic transplantation will undoubtedly occur and such trials have been initiated at several institutions. Careful attention to vector, suicide gene, selectable marker, efficiency of transduction, and cell dose will be necessary when comparing different trials since these variables will probably affect transgenic cell survival and response rates. [figure: see text] PMID- 10800658 TI - Clinical application of hematopoietic stem cell culture and expansion. PMID- 10800659 TI - New cytokines and their clinical application. PMID- 10800660 TI - Inherited risk factors for venous thromboembolism: implications for clinical practice. AB - Each year, thrombosis in the deep veins of the lower extremities occurs in 1 of every 1000 Americans (1). Although most cases of venous thrombosis are not fatal, death from pulmonary embolism can be expected in 1% to 2% of all patients, and as many as 25% of patients with deep venous thrombosis (DVT) will suffer the chronic effects of postthrombotic syndrome. Thus, identification of high-risk patients with genetic predispositions to thrombosis is an important clinical goal, particularly among individuals with recurrent DVT. PMID- 10800661 TI - Prophylaxis for deep venous thrombosis and pulmonary embolism in the surgical patient. AB - The fifth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy provides the most up-to-date guidelines for the prevention of deep venous thrombosis (DVT) and pulmonary embolism (PE) in the surgical patient (1). These recommendations have become a major guideline for clinicians managing patients in the perioperative period. Despite these recommendations, there remains a concern for balancing the risk of major postoperative bleeding with the benefit of preventing thrombosis. In an attempt to resolve this issue, clinicians have requested clear-cut guidelines for identification of high-risk groups for whom prophylaxis must be used. This article will review the etiology, risk-factor stratification, regimens of prophylaxis, and recommendations for prevention of postoperative DVT and PE. PMID- 10800662 TI - Diagnosis of deep venous thrombosis. AB - Although deep venous thrombosis (DVT) is often considered an elusive disease that is difficult to recognize clinically, important clinical clues and properly ordered imaging tests can reliably detect or exclude this condition. This article will provide a practical approach to recognizing clinical settings in which DVT should be suspected, suggesting a focused history and physical examination and outlining optimal strategies for test ordering. This information will help ensure that patients with DVT are identified rapidly and, conversely, that those without DVT will receive reassurance or specific treatment for the medical problem that mimics it. Once DVT is suspected, a workup can proceed rapidly and accurately to resolve the diagnostic issue. PMID- 10800663 TI - Diagnosis of pulmonary embolism. AB - Although pulmonary embolism (PE) is often considered a difficult diagnosis to establish, important clinical clues and properly ordered laboratory and imaging tests can reliably detect or exclude this potentially fatal condition. This article will provide a practical approach to recognizing clinical situations in which PE should be suspected. This guide to PE diagnosis will be tailored to those with busy practices, outlining a focused history and physical examination as well as optimal strategies for test ordering. Instead of performing an encyclopedic laundry list of tests, clinicians can pick and choose carefully among them, which maximizes efficiency and cost-effectiveness. The choice of tests is flexible, thus allowing the clinician to utilize the strengths of the particular hospital in which the patient is evaluated. This article will help ensure that patients with PE are identified rapidly and, conversely, that those without PE will undergo an appropriate workup for other important medical conditions. With a 3-stage approach consisting of (1) clinical evaluation; (2) nonimaging laboratory tests; and (3) imaging tests, PE can often be quickly detected with a definitive evaluation. PMID- 10800664 TI - Management of deep venous thrombosis and pulmonary embolism. AB - The management of deep venous thrombosis (DVT) and pulmonary embolism (PE) is changing dramatically. The US Food and Drug Administration has approved outpatient treatment of DVT with the low-molecular-weight heparin enoxaparin as a bridge to warfarin. Warfarin use is improved by avoiding loading doses and by recognizing previously unappreciated interactions and potentiation with commonly used medications such as acetaminophen. The importance of isolated calf and upper extremity venous thromboses has been validated, so that patients with these conditions routinely undergo anticoagulation. Risk stratification for PE is becoming more sophisticated because practitioners now assess right ventricular function (usually by echocardiography) instead of relying solely on systemic arterial blood pressure and heart rate to determine prognosis. Among patients with massive DVT or hemodynamically unstable PE, thrombolysis, thrombectomy, and embolectomy (often performed in an interventional angiography laboratory) are being used with increasing skill and improved outcomes. PMID- 10800666 TI - Analysis of protein-protein interactions in phototransduction cascade using surface plasmon resonance. PMID- 10800665 TI - Inhibition of rhodopsin phosphorylation by S-modulins: purification, reconstitution, and assays. PMID- 10800667 TI - Expression of phototransduction proteins in Xenopus oocytes. PMID- 10800668 TI - Xenopus rod photoreceptor: model for expression of retinal genes. PMID- 10800670 TI - Bovine retinal nucleoside diphosphate kinase: biochemistry and molecular cloning. PMID- 10800671 TI - Calcium-binding proteins and their assessment in ocular diseases. PMID- 10800669 TI - Fusion between retinal rod outer segment membranes and model membranes: functional assays and role for peripherin/rds. PMID- 10800672 TI - Molecular structure of membrane-targeting calcium sensors in vision: recoverin and guanylate cyclase-activating protein 2. PMID- 10800673 TI - Measurement of light-evoked changes in cytoplasmic calcium in functionally intact isolated rod outer segments. PMID- 10800674 TI - Laser spot confocal technique to measure cytoplasmic calcium concentration in photoreceptors. PMID- 10800675 TI - Functional study of rhodopsin phosphorylation in vivo. PMID- 10800676 TI - Mechanisms of single-photon detection in rod photoreceptors. AB - Rod photoreceptors detect and encode incident photons exceptionally well. They collect sparse photons with high efficiency, maintain a low dark noise, and generate reproducible responses to each absorbed photon. The mechanisms involved in single-photon detection--control of the effective lifetime of a single active receptor molecule, amplification of the activity of this single molecule by a second-messenger cascade, and reliable transmission of small synaptic signals- recur throughout the nervous system. Indeed, several other sensory systems reach or approach limits set by quantization of their input signals. For example, olfactory receptors can detect single odorant molecules. Although our understanding of visual transduction and signal processing has advanced rapidly during the past 10-15 years, fundamental questions still remain: What mechanisms are responsible for the reproducibility of the rod's elementary response? What are the tradeoffs of speed and sensitivity in the transduction cascade? How are the rod single-photon responses reliably transmitted to the rest of the visual system? Future technical innovations, particularly better methods to monitor the activity of intermediate steps in transduction, will play an important role in providing answers. PMID- 10800677 TI - Electroretinographic determination of human rod flash response in vivo. PMID- 10800678 TI - Electrophysiological methods for measurement of activation of phototransduction by bleached visual pigment in salamander photoreceptors. PMID- 10800679 TI - Exploring kinetics of visual transduction with time-resolved microcalorimetry. PMID- 10800680 TI - Use of alpha-toxin-permeabilized photoreceptors in in vitro phototransduction studies. PMID- 10800681 TI - Photoreceptors in pineal gland and brain: cloning, localization, and overexpression. PMID- 10800682 TI - Cultured amphibian melanophores: a model system to study melanopsin photobiology. PMID- 10800683 TI - High-performance liquid chromatography analysis of visual cycle retinoids. PMID- 10800684 TI - Quantitative measurements of isomerohydrolase activity. PMID- 10800686 TI - Analyzing membrane topology of 11-cis-retinol dehydrogenase. PMID- 10800685 TI - Multienzyme analysis of visual cycle. PMID- 10800687 TI - Phase partition and high-performance liquid chromatography assays of retinoid dehydrogenases. PMID- 10800688 TI - Short-chain dehydrogenases/reductases in retina. PMID- 10800689 TI - Substrate specificities of retinyl ester hydrolases in retinal pigment epithelium. PMID- 10800691 TI - Analysis of chromophore of RGR: retinal G-protein-coupled receptor from pigment epithelium. PMID- 10800690 TI - Molecular characterization of lecithin-retinol acyltransferase. PMID- 10800692 TI - Low-temperature photoaffinity labeling of rhodopsin and intermediates along transduction path. PMID- 10800693 TI - Structural analysis of protein prenyl groups and associated C-terminal modifications. PMID- 10800694 TI - Isoprenylation/methylation and transducin function. AB - Freshly prepared proteolyzed (deprenylated) T beta gamma and material isolated from retina are inert with respect to activating T alpha in the presence of R* in detergent and in disk membranes. In addition, proteolyzed T beta gamma is also incapable of supporting the pertussis toxin-catalyzed ADP ribosylation of T alpha GDP. These experiments show that isoprenylation/methylation is essential for the fruitful interactions between T alpha and T beta gamma at the membrane. When tested for its ability to support GTP-for-GDP exchange catalyzed by R*, demethylated T beta gamma proved to be approximately 50% as active as methylated T beta gamma in photoreceptor disk membranes (Fig. 3) and in reconstituted liposomes containing rhodopsin. In detergent, no difference was observed between methylated and demethylated T beta gamma, suggesting no role at all for the methyl group in functional interactions between T alpha, T beta gamma, and R*. The twofold activity difference observed in membranes can be accounted for by the twofold lessened affinity of the demethylated T beta gamma, compared with its methylated counterpart, for membranes in the presence of R* and T alpha. It is interesting to note that a substantially larger difference (> 10-fold) in the relative binding of methylated versus demethylated T beta gamma to membranes is observed in the absence of R* and T alpha. However, R* has a substantial affinity for T alpha beta gamma, and the influence of R* and T alpha greatly reduces any differences resulting from the presence or absence of a methyl group on T beta gamma. The results from studies of demethylated T beta gamma demonstrate that specific lipid-receptor interactions are unlikely to play a critical role in the rhodopsin-transducin system, and further show that the effect of methylation is probably due to the increased hydrophobicity of methylated T beta gamma versus its unmethylated counterpart. These studies are, of course, relevant to heterotrimeric G proteins, and specifically to the interactions of receptor (R*) with T alpha and T beta gamma. If a hydrophobic lipid-lipid mechanism is operative, the state of methylation would be expected to have a more profound effect on the membrane-associative properties of farnesylated proteins, but not on those of geranylgeranylated proteins. The increased hydrophobicity of the C20 geranylgeranyl group relative to the C15 farnesyl group will compensate for the loss of the methyl substituent. The results obtained in the transducin-rhodopsin system can be contrasted with the effect of gamma-subunit methylation on effector enzyme activation. In the case of the geranylgeranylated beta 1 gamma 2, methylation proved to have only a small effect on PIPLC beta activation (Fig. 4B). An approximately 25% diminution in efficacy, but not potency, was observed for the demethylated geranylgeranylated beta 1 gamma 2 versus its methylated counterpart. This again shows that specific lipid-protein interactions are unimportant. The effect of methylation on membrane binding would be expected to be small, given that beta 1 gamma 2 is geranylgeranylated. It is of interest to compare these results with those found with methylated and unmethylated T beta gamma as activators of PIPLC beta. In this instance there was a large effect noted, with methylated T beta gamma being at least 10-fold more potent than its unmethylated counterpart with respect to activating either enzyme (Fig. 4A). This result is readily understandable in light of the role of methylation in selectively enhancing hydrophobicity of farnesylated proteins as opposed to geranyl-geranylated proteins. Similar results were obtained for the activation of PI3K, further strengthening the conclusion that it is lipid-lipid interactions that direct beta gamma subunit membrane association. (ABSTRACT TRUNCATED) PMID- 10800695 TI - Functional analysis of farnesylation and methylation of transducin. PMID- 10800696 TI - Identification of phosphorylation sites within vertebrate and invertebrate rhodopsin. PMID- 10800697 TI - Ocular proteomics: cataloging photoreceptor proteins by two-dimensional gel electrophoresis and mass spectrometry. PMID- 10800698 TI - Animal models of inherited retinal diseases. PMID- 10800699 TI - Avian models of inherited retinal disease. PMID- 10800701 TI - Genetic models to study guanylyl cyclase function. PMID- 10800700 TI - Biochemical analysis of phototransduction and visual cycle in zebrafish larvae. PMID- 10800703 TI - In situ hybridization studies of retinal neurons. PMID- 10800702 TI - Analysis of visual cycle in normal and transgenic mice. PMID- 10800704 TI - Cloning and characterization of retinal transcription factors, using target site based methodology. PMID- 10800705 TI - In vivo assessment of photoreceptor function in human diseases caused by photoreceptor-specific gene mutations. PMID- 10800706 TI - Spectral sensitivities of human cone visual pigments determined in vivo and in vitro. PMID- 10800707 TI - Molecular analysis of human red/green visual pigment gene locus: relationship to color vision. PMID- 10800708 TI - Expression and characterization of peripherin/rds-rom-1 complexes and mutants implicated in retinal degenerative diseases. AB - Nearly 40 disease-linked mutations have been reported for peripherin/rds to date; heterologous expression in tissue culture cells offers a valuable means of efficiently characterizing the biochemical properties of the various mutants. Peripherin/rds is proposed to act as an essential structural element in outer segment disk morphogenesis, and a present transgenic mice offer the sole tractable system in which recombinant peripherin/rds may be examined functionally in situ. Because the generation and characterization of transgenic animals are both expensive and time consuming, heterologous expression in cultured cells offers an important and complementary means of addressing protein structure and function. The immunopurification and detection of the peripherin/rds-rom-1 complex are performed using specific immunochemical reagents, monoclonal and polyclonal antibodies, that are not commonly available. Several laboratories have developed antibodies to peripherin/rds and rom-1 in rabbits and mice, using a variety of immunogens: purified ROS membranes, purified E. coli fusion proteins, and synthetic peptides coupled to proteins. The C-terminal regions appear to be most highly antigenic, although antibodies have been generated to other regions as well. Regardless of their source, antibodies must be thoroughly characterized; specificity is often a function of solution conditions and must be determined empirically. The approach as described here has provided explanations for several instances of peripherin/rds-associated disease, including digenic RP linked to as L185P mutation, and adRP associated with C118/119del and C214S mutations. In addition, the R172W mutation, linked to macular dystrophy and preferential loss in cone function, is shown to behave normally with respect to biosynthesis and subunit assembly; it likely involves a more subtle functional defect that remains to be described. Finally, the methodology reported here has suggested the existence of a novel (homotetrameric) form of peripherin/rds in individuals lacking rom-1; this hypothesis has been confirmed in rom-1 knockout mice. The information obtained thus far demonstrates the utility of using heterologously expressed peripherin/rds and rom-1 to investigate the consequences of disease linked mutations in these polypeptides. Heterologous cell expression coupled with transgenic mouse methodologies should continue to provide a more detailed understanding of molecular mechanisms underlying inherited retinal degenerative diseases. PMID- 10800709 TI - Isolation of retinal proteins that interact with retinitis pigmentosa GTPase regulator by interaction trap screen in yeast. PMID- 10800710 TI - Genetic analysis of RPE65: from human disease to mouse model. PMID- 10800711 TI - Construction of encapsidated (gutted) adenovirus minichromosomes and their application to rescue of photoreceptor degeneration. PMID- 10800712 TI - Production and purification of recombinant adeno-associated virus. PMID- 10800713 TI - Ribozymes in treatment of inherited retinal disease. PMID- 10800714 TI - Cross-species comparison of in vivo reporter gene expression after recombinant adeno-associated virus-mediated retinal transduction. PMID- 10800715 TI - Separation of carrier-free ytterbium and thulium produced in 80 MeV 12C6+ irradiated gadolinium foil target by liquid-liquid extraction with HDEHP. AB - Heavy ion activation of natural gadolinium foil with 80 MeV 12C6+ results in the formation of carrier-free ytterbium isotopes, 165,166,167Yb and their corresponding decay products, 165,166,167Tm, in the matrix. Carrier-free ytterbium and thulium isotopes have been separated quantitatively from the bulk target matrix gadolinium by employing di-(2-ethylhexyl)phosphoric acid (HDEHP) as a liquid cation exchanger. PMID- 10800716 TI - Effects of soil acidity on the uptake of trace elements in soybean and tomato plants. AB - The effects of soil acidity on the uptake of trace elements (Co, Zn, Se, Rb, Sr, Y, Zr, Tc, Ru, Rh and Re) in soybean and tomato were studied by a multitracer technique. The soybean and tomato plants were cultivated on soils at pH 6.4 (normal soil) and 4.2 (acid soil) and administered with a multitracer for 15-60 d. In general, the uptake of cationic elements in the leaves and stems of soybean plants cultivated on acid soil became higher than those of plants cultivated on normal soil during the late period of growth. However, the effect of soil acidification on the uptake of the anionic element. Se, was quite different from that on the cationic elements. The uptake of Se by the plants cultivated on normal soil was higher than that of the plants cultivated on acid soil at all four harvest points. The uptake behavior of these elements in soybean was discussed in relation to their adsorption behavior on the same soil as was used for soybean cultivation. The growth of tomato plants was seriously affected by the soil acidity and lowering of uptake of elements was observed for the plants cultivated on acidified soil. PMID- 10800717 TI - Sorption and desorption of radiocesium on red earth and its solid components: relative contribution and hysteresis. AB - The relative contributions of organic matter and iron oxides to the Cs+ sorption on red earth were investigated by using the batch technique and selective extraction methods. The sorption and desorption isotherms and the distribution coefficients of Cs+ on the untreated red earth and the three treated soils to remove organic matter, iron oxides and organic matter plus iron oxides were determined at 20 degrees C, pH 6.3+/-0.2, in the presence of 0.01 mol/l CaCl2. It was found that all the isotherms are linear in the Cs+ concentration range used here, that the sorption desorption hysteresis on the red earth actually occurs, and besides the clay minerals, the organic matter present in the red earth is a significant trap of Cs+ and is responsible for the hysteresis instead of the iron oxides. PMID- 10800718 TI - Adsorption characteristics of 47 elements on a calcareous soil, a red earth and an alumina: a multitracer study. AB - The distribution coefficients, Kd of 47 elements from Na to Bi on a calcareous soil, a red earth and an alumina were, respectively, determined using a multitracer technique. The multitracer solution was prepared by irradiation of UO2(NO3)2 with 40Ar ion beam. It was found that there is a common general trend of variation in Kd for both soils and the alumina, i.e. all the shapes of plots of log Kd versus electron binding energy (Iz) are an 'inverted V', hence the important role of chemical interaction in the adsorption of trace inorganic cations, anions and neutral complexes on the soils and the oxides was demonstrated. For the alkali and alkaline earth elements, the common sequences of Kd values on the three adsorbents were Na+ < K+ < Rb+ < Cs+ (except for Rb+ on the alumina) and Mg2+ < Sr2+ < Ba2+ and accord with the sequences of effective ionic radii or single ion hydration enthalpies. PMID- 10800719 TI - Retrospective dosimetry: estimation of the dose to quartz using the single aliquot regenerative-dose protocol. AB - We report on the application of the single-aliquot regenerative-dose protocol to retrospective dosimetry, using the optically stimulated luminescence (OSL) from quartz extracted from fired bricks. These bricks had previously been exposed to enhanced levels of ionising radiation while part of inhabited structures in the Chernobyl area. The time dependence of the OSL signals is considered first and we conclude that it is most appropriate to use the initial part of the OSL signal for dose estimation, after subtraction of a slowly varying background component. It is then shown that this signal meets the fundamental requirement of the single aliquot regenerative-dose protocol, in that any change in the luminescence recombination probability can be corrected for by using the OSL response to a fixed test dose. The response of a particular aliquot is examined after three different treatments (untreated, reset by exposure to light and reset by heating to 500 degrees C) and it is shown that, after sensitivity correction, the dose response curves are indistinguishable up to 10 Gy. The routine application of the protocol is then described and dose estimates are shown to be insensitive to preheat temperature and test-dose size. Finally, dose-depth profiles are presented for two bricks. These profiles demonstrate that the high precisions (approximately 1%) obtained using the regenerative-dose protocol are reflected in smooth dose-depth dependencies. PMID- 10800720 TI - Development of an automated synthesis apparatus for L-[3-11C] labeled aromatic amino acids. AB - We have developed an automated synthesis apparatus for L-[3-11C]-labeled aromatic amino acids by improved multi-enzymatic reactions. Use of a specially designed reaction vessel and of CH2Cl2 as a solvent has greatly simplified the overall operation, proving to be suitable for automated synthesis, and also has given better yields and higher specific activities than formerly reported values. PMID- 10800721 TI - Preparation of rhenium-188-tin colloid as a radiation synovectomy agent and comparison with rhenium-188-sulfur colloid. AB - As a generator-produced beta-emitting radionuclide, the importance of 188Re for radionuclide therapy is increasing rapidly. We prepared 188Re-tin colloid and compared its properties with 188Re-sulfur colloid. Labeling efficiencies reached >98% for tin colloid at 2 h and 89-94% for sulfur colloid at 3 h. All the preparations were stable for 72 h in water, serum, and synovial fluid. If labeled at higher temperature, the particle size of tin colloid increased. The residual radioactivity of 188Re-sulfur colloid in disposable polypropylene syringes after injecting mice was high (62.0+/-7.0%) due to its hydrophobic nature, while that of 188Re-tin colloid was low (2.9+/-1.6%). Although both 188Re-tin colloid and 188Re-sulfur colloid might be useful for radionuclide therapy, we conclude that 188Re tin colloid is more advantageous over 188Re sulfur colloid, due to higher labeling efficiency, control of the particle size, and lower residual activity in the injection syringes. PMID- 10800722 TI - Synthesis and biodistribution of [11c]procaterol, a beta2-adrenoceptor agonist for positron emission tomography. AB - The potent, subtype-selective radioligand (+/-)-erythro-5-(1-hydroxy-2 [11C]isopropyl-aminobutyl)-8-hydroxy-car bostyril ([11C]procaterol) was synthesized and evaluated for visualization of pulmonary beta2-adrenoceptors with positron emission tomography (PET). Procaterol was labelled by reductive alkylation of the desisopropyl precursor with [11C]acetone under the influence of NaCNBH3 and acetic acid. Synthesis and HPLC purification were performed in 34 min. Specific activities ranged from 26.5-39.3 TBq (about 700-1000 Ci)/mmol and the radiochemical yield was 2.4-8.6% (corrected for decay). Biodistribution studies were performed in male Wistar rats which were either untreated or predosed with (D,L)-propranolol hydrochloride (beta-adrenoceptor antagonist, 2.5 mg/kg), ICI 118551 (beta2-adrenoceptor antagonist, 0.15 mg/kg), CGP 20712A (beta1 adrenoceptor antagonist, 0.15 mg/kg) or isoprenaline (beta1-adrenoceptor agonist, 15 mg/kg). Specific binding was observed in lungs, spleen and red blood cells, tissues known to contain beta2-adrenoceptors. Pulmonary binding was blocked by propranolol, ICI 118551 and isoprenaline, but not by CGP 20712A. This binding pattern is consistent with the beta2 selectivity of the radioligand. The clearance of [11C]procaterol was biphasic, with a rapid distribution phase (t1/2 0.17 min) representing 90% of the injected dose followed by an elimination phase (t1/2 18.1 min). About 45% of the plasma radioactivity was unmetabolized procaterol at 15 min postinjection. In a dynamic PET-study, the lungs of untreated control rats could barely be detected and total/non-specific binding ratios rose to only 1.2 at 20 min postinjection. Although labelling and administration of (-) erythroprocaterol, the most active of 4 stereoisomers, may produce better results, [11C]procaterol seems unsuitable for beta-adrenoceptor imaging. PMID- 10800723 TI - Calibration of a field-portable gamma detector to obtain in situ measurements of the 137Cs inventories of cultivated soils and floodplain sediments. AB - Over the past 10 years, a number of studies have exploited the potential for using measurements of fallout 137Cs inventories to document rates and patterns of soil erosion on cultivated land and to estimate rates of overbank sedimentation on river floodplains. Traditional procedures for applying the 137Cs technique involve the collection of soil or sediment cores from a study site and their subsequent transfer to the laboratory for preparation and analysis by gamma spectrometry. Such procedures are time consuming and there may be a considerable delay before the results are available. It is therefore difficult to obtain preliminary results, which could be used to guide the development of an ongoing sampling programme. The use of in situ gamma spectrometry measurements to quantify 137Cs inventories in soils and sediments offers a number of potential advantages over traditional procedures. However, in order to derive a reliable estimate of the 137Cs inventory for a measurement point, it is necessary to take account of the attenuation of 137Cs gamma rays by the soil matrix and information on the depth distribution of 137Cs in the soil or sediment is therefore required. In the present study, empirical relationships between in situ measurements of 137Cs activity and total 137Cs inventories have been established for soils from a cultivated field and for floodplain sediments, based on information on the vertical distribution of 137Cs in the soils and sediments provided by the forward scattering ratio derived from the field measured spectra. These relationships have been used to estimate 137Cs inventories from in situ measurements of 137Cs activity at other locations. PMID- 10800724 TI - Excitation of the 180mTa isomer in (gamma,n) reactions and its astrophysical relevance AB - The yield has been determined for the excitation of 180gTa (Jpi = 1+) from the 181Ta(gamma,n) reaction by measurements of gamma rays emitted following the electron capture and beta-decays of the 8.15 h ground state. The probability sigmam/(sigmam + sigmag) for the production of the 180mTa isomer (Jpi = 9-) after gamma absorption in the energy region from 8 to 13 MeV was deduced. The role of the initial and final spins on the reaction yield, along with the relevance for nucleosynthesis of 180mTa, is discussed. PMID- 10800725 TI - Standardization and photon emission probabilities in the decay of 237Np/233Pa AB - The activity concentration of a 237Np solution was determined by 4pialpha counting, by alpha particle spectroscopy with defined solid angle and by liquid scintillation counting (LSC). These methods yielded results which differ by less than 0.1% from the mean, the relative standard uncertainties being 0.23% (4pialpha), 0.27% (LSC) and 0.5% (solid angle). X-ray and gamma-ray emission probabilities of several transitions were measured using semiconductor detectors. PMID- 10800726 TI - A method for the selection of the optimum counting conditions in a ZnS(Ag) scintillation detector. AB - The well-known criteria for the selection of the optimum counting conditions in a ZnS(Ag) scintillation detector seem to restrict its applicability. These do not consider simultaneously operating voltage and electronic threshold variation to reach the best counting conditions. Therefore, a more general method for the determination of the optimum counting conditions in ZnS(Ag) scintillation detectors is proposed. In this method, a relationship between voltage and electronic threshold is derived for counting efficiency. In order to test the method, quality control procedures have been carried out as well as the determination of gross alpha activity in environmental samples. PMID- 10800727 TI - Determination of the excitation function for the 10B(p,n)10C reaction with implications for the production of [10C]carbon dioxide for use as a PET tracer. AB - To evaluate the possibilities of producing useful amounts of 10C (T1/2 = 19 s) for use as an on-line, steady state positron emission tomography (PET) tracer, the excitation function and the thick target yield for the 10B(p,n)10C reaction up to 30 MeV were measured using an adapted version of the stacked-foil technique. The radionuclidic purity of the produced 10C versus the inevitable 11C contamination was evaluated as function of target thickness and incident proton beam energy. PMID- 10800728 TI - The distribution of 137Cs and 90Sr in the biomass of pine trees planted in 1987 1988 in the near zone of the Chernobyl nuclear power plant. AB - This paper presents the results of a study, carried out in 1996, of the distributions of 137Cs and 90Sr in the biomass of a new pine plantation, planted in 1987-1988, in the near field of the Chernobyl nuclear power plant. The maximum observed concentrations were 4.2 MBq kg(-1) for 137Cs in the youngest needles and 1.1 MBq kg(-1) for 90Sr in the oldest needles. The estimation of the total absorbed doses in the different organs of the pine was also carried out, the observed doses were in the range 2 Gy yr(-1) to 16.8 Gy yr(-1); the lifetime dose for the needles (4 yr) was 44 Gy. PMID- 10800729 TI - Generic models for radionuclide dosimetry: 11C-, 18F- or 75Se-labelled amino acids. AB - A generic biokinetic model was developed for use in the assessment of the internal dose received by human subjects injected with amino acids labelled with 11C (T1/2 = 0.34 h, beta+, gamma), 18F(T1/2 = 1.83 h, beta+, gamma) or 75Se (T1/2 = 119.8 d, beta-, gamma). This generic model was used in conjunction with the MIRDOSE 3 computer programme to calculate radiation doses to adults; these radiation doses were compared with those calculated using compound-specific models for two [11C]-, and eight [18F]-labelled amino acids and [75Se]selenomethionine. In general, the effective doses, as well as the organ and tissue doses, calculated using the generic model agreed within a factor of 2 or less, with those calculated using compound-specific models; the generic model tended to over-, rather than underestimate the organ and tissue doses. It was concluded that for 11C- and 18F-labelled amino acids and for 75Se-labelled amino acids or their analogues, the single generic biokinetic model could be applied for general radiation protection purposes. PMID- 10800730 TI - Excitation functions of 120Te(d,xn)121,120m,gI reactions from threshold up to 13.5 MeV: comparative studies on the production of 120gI. AB - Excitation functions of the nuclear reactions 120Te(d,xn)121,120m,gI were measured for the first time from their respective thresholds up to 13.5 MeV. Thin samples prepared by electrolytic deposition of 99.0% enriched 120Te on Ti-backing were used. Integral yields of 121,120m,gI were calculated from the measured cross section data. A comparison of the 122Te(p,3n)-, 120Te(p,n)- and 120Te(d,2n) processes for the production of 120gI is given. The 120Te(d,2n)-process is unsuitable for production purposes since the yield of 120gI is very low and the level of 121I impurity very high. The choice lies either on the 122Te(p,3n)- or the 120Te(p,n)-reaction and is governed by the available proton energy and the financial resources for procuring the enriched target material. PMID- 10800731 TI - Ultrafiltration of large volumes for the determination of colloidally bound artificial radionuclides in natural waters. AB - A methodology based on large volume ultrafiltration (> 1000 l) is developed in order to characterize the artificial radionuclides associated with the colloidal populations in natural waters. "Inorganic elements" and organic carbon sinks or sources due to sorption or post desorption within the ultrafiltration assembly/membranes of the Sartorius system are underlined and discussed from methodological step experiments as well as recoveries obtained for artificial radionuclides such as 106Ru, 125Sb, 137Cs and 239+240Pu fractionating natural waters from the Rhone River (France), its estuary and the Mediterranean Sea (Gulf of Lions). PMID- 10800732 TI - Nuclear model calculations on proton and deuteron induced reactions on 122Te and 120Te with particular reference to the formation of the isomeric states 120m,gI. AB - Nuclear model calculations were performed on (p,xn) reactions on 122Te and 120Te, and (d,xn) reactions on 120Te. The computer code used (STAPRE) incorporates statistical and precompound models as well as nuclear structure effects. The total cross-sections of (p,xn) and (d,xn) processes leading to the formation of the radioisotopes 119I, 120gI, 121I and 122I are described well by the calculations. The isomeric cross-section, however, is rather difficult to calculate. The yield of the high spin isomer (120mI) depends on the type of reaction involved and increases with the increasing projectile energy. From an analysis of the energy dependence of the isomeric cross-section ratio, the excitation energy of 120mI was deduced to be 550+/-50 keV and its spin and parity as 4+. The experimental data reported earlier and the theoretical analysis presented in this work allow to define the optimum conditions for the production of the medically important beta+ emitter 120gI with enhanced confidence. PMID- 10800733 TI - Resolution of a position-sensitive proportional-counter with a resistive anode wire of carbon fiber AB - Theoretical and experimental analyses have been performed to examine the position resolution of a proportional counter with a 7-microm diameter carbon fiber as a resistive anode, which is used to encode one-dimensional positions of incident photons according to the charge-division method. These analyses have clearly shown that the carbon fiber is applicable to the position sensing of the order of 100-200 microm as long as the filled gas is pressurized at near 10 atm. PMID- 10800734 TI - Redetermination of the total internal conversion coefficient of the 279 keV transition following the decay of 203Hg AB - A new measurement of the total internal conversion coefficient of the 279 keV transition following the decay of 203Hg resulted in alpha = 0.2250(12). PMID- 10800735 TI - Evidences of precession and obliquity orbital forcing in oxygen-18 isotope composition of Montalbano Jonico section (Basilicata, southern Italy) AB - Quantitative signal processing methods have been applied to a delta(18)O profile for a land-based stratigraphic section, extending from the upper part of lower Pleistocene to the lower part of middle Pleistocene. The section is well exposed with a continuous succession of muds and muddy silts, about 400 m thick, located in the southernmost part of Bradano Trough, near Montalbano Jonico in Basilicata (south Italy). The sampled part of the section is about 240 m thick, in which a foram benthic species (Cassidulina carinata) is continuously available for oxygen isotope ratio measurements. The aim of the data treatment is to discover how much of the Earth's orbital periodic movements, precession and obliquity, which represent the dominant periodicities in paleoclimatic variations from the base of the Pleistocene until 0.735 Myr BP, are responsible for the oscillations observed in the oxygen-18 record of the Montalbano Jonico section. A time framework of the section was constructed on the basis of calcareous nannofossil biostratigraphic analyses, preliminary magnetostratigraphic results and oxygen isotope correlation with the record from DSDP s607 (isotope data collected in the NOAA World Data Center). The resulting time-scale extends from 1.15 to 0.74 Myr. Power spectrum analysis was performed on the isotope data to illuminate the most important periodicity components of the Montalbano Jonico record. The periodic components of 41,000 and 21,000 yr are present in this record; the former associated with periodic changes in the tilt of Earth's axis and the latter with periodic changes with the precession of the equinoxes, as predicted by the astronomical theory of ice ages. They are, however, not the most important components of the power spectrum, in which a lower frequency component contains most of the variance. This low-frequency component is centered at a period around 208,000 yr. This periodicity seems not to be attributable to any known astronomical or paleoclimatic phenomenon. An attempt was made to verify if this periodicity was due to the composite effect of precession and obliquity signals together at different frequencies from their forcing frequencies. In order to investigate this effect, isotope data have been parameterized in terms of a sum of simple functions of precession and obliquity signals with unknown coefficients. The coefficients are estimated from the time series with the assumption that the best coefficients are those which minimize the 'noise' i.e. the difference between the data function and the precession and obliquity functions. Cross-spectra analyses were also performed on the data and the precession signal and on the data and the obliquity signal. The power spectrum of the residual 'noise' functions and the cross-spectra demonstrate that precession and obliquity signals are not in phase with the data at their forcing frequencies and so damp. The precession and obliquity signals were then shifted towards lower frequencies at equally spaced lags, the resultant 'noise' power spectra were plotted for every combination of lags of precession and obliquity. The results of this data processing demonstrate that it is possible to have a combination of precession and obliquity cyclicities that could be responsible for the signal with 208 kyr periodicity. PMID- 10800736 TI - The transfer of uranium isotopes 234U and 238U to the waters interacting with carbonates from Mendip Hills area (England). AB - Laboratory time-scale experiments were conducted on limestone and dolomite gravels from the Mendip Hills area, England, with the purpose of evaluating the release of 238U and 234U to different aqueous solutions. The 234U/238U activity ratio (AR) lab data were reliable to interpret the field data. The obtained values do not indicate a reduction in the amount of dissolved U and an increase in the AR of the remaining dissolved U as commonly observed for groundwater systems close to redox boundaries. PMID- 10800737 TI - Helium flux from a sedimentary basin. AB - The tritium-3He ages of groundwaters collected from the Saijo Basin in Japan were measured. The ages vary between 11.7 and 16.3 years. The 4He flux of >3 x 10(4) atoms/cm2 s is calculated by the 4He concentrations and ages. In addition the helium flux of 8.3 x 10(5) atoms/cm2 s is calculated at the Higashi-Niigata gas field in Japan. The above two estimates are consistent with the continental helium flux reported in the literature, suggesting that helium flux on the Earth's crust is uniform. PMID- 10800738 TI - A probabilistic description of radioactive contamination. AB - A discrete probability model is used to aid in the description of gamma-ray spectroscopic data obtained from a radioactively contaminated territory. The approach gives a natural probability for distinguishing contaminated regions and is useful for describing spatial vector valued discrete random fields. Using the simplest variant of this model, a part of the territory adjoining the former Semipalatinsk's nuclear test site in Kazakhstan has been investigated. Probabilities of exceeding the safety limits for 137Cs and integral gamma radiation have been estimated. PMID- 10800739 TI - Comparison of uranium determination in some Syrian geologic samples using three reactor based methods AB - A set of 25 samples of soil, sediments, carbonate and phosphate rocks from Syria were analysed for uranium, using three reactor based methods; instrumental neutron activation analysis (INAA), delayed neutron counting (DNC) and one cycle of irradiation utilizing the cyclic activation system (CAS). Although the three methods are capable of irradiation samples, the last method is the least established for U determination in rocks. The measurements obtained by the three methods are compared. The results show good agreement, with a distinct linear relationship and significant positive correlation coefficients. It was concluded that the CAS method could reliably be used to rapidly determine uranium in geological samples. PMID- 10800740 TI - Uranium-thorium levels in the sediments of the Kubanni River in Nigeria. AB - U-Th levels of the Kubanni River sediments in the Nigerian Basement Complex have been determined and shown to be enriched with mean values of 9.06+/-2.66 and 21.44+/-4.12 (ppm) respectively. The probable contributory factors are either phosphate fertilisers used on the neighbouring farms or low aeolian depositions from the NE trade winds blowing from the Sahara desert across the Northern Nigeria Savannah region. The other major contributing factor is the annual weathering of the basement granites. This work revealed more clustering of our data sets below the igneous line as proposed by other authors. PMID- 10800741 TI - A "theory of relativity" for cognitive elasticity of time and modality dimensions supporting constant working memory capacity: involvement of harmonics among ultradian clocks? AB - 1. The capacity of working memory (WM) for about 7+/-2 ("the magical number") serially organized simple verbal items may represent a fundamental constant of cognition. Indeed, there is the same capacity for sense of familiarity of a number of recently encountered places, observed in radial maze performance both of lab rats and of humans. 2. Moreover, both species show a peculiar capacity for retaining WM of place over delays. The literature also describes paradoxes of extended time duration in certain human verbal recall tasks. Certain bird species have comparable capacity for delayed recall of about 4 to 8 food caches in a laboratory room. 3. In addition to these paradoxes of the time dimension with WM (still sometimes called "short-term" memory) there are another set of paradoxes of dimensionality for human judgment of magnitudes, noted by Miller in his classic 1956 paper on "the magical number." We are able to reliably refer magnitudes to a rating scale of up to about seven divisions. Remarkably, that finding is largely independent of perceptual modality or even of the extent of a linear interval selected within any given modality. 4. These paradoxes suggest that "the magical number 7+/2" depends on fundamental properties of mammalian brains. 5. This paper theorizes that WM numerosity is conserved as a fundamental constant, by means of elasticity of cognitive dimensionality, including the temporal pace of arrival of significant items of cognitive information. 6. A conjectural neural code for WM item-capacity is proposed here, which extends the hypothetical principle of binding-by-synchrony. The hypothesis is that several coactive frequencies of brain electrical rhythms each mark a WM item. 7. If, indeed, WM does involve a brain wave frequency code (perhaps within the gamma frequency range that has often been suggested with the binding hypothesis) mathematical considerations suggest additional relevance of harmonic relationships. That is, if copresent sinusoids bear harmony-like ratios and are confined within a single octave, then they have fast temporal properties, while avoiding spurious difference rhythms. Therefore, if the present hypothesis is valid, it implies a natural limit on parallel processing of separate items in organismic brains. 8. Similar logic of periodic signals may hold for slower ultradian rhythms, including hypothetical ones that contribute to time-tagging and fresh sense of familiarity of a day's event memories. Similar logic may also hold for spatial periodic functions across brain tissue that, hypothetically, represent cognitive information. Thus, harmonic transitions among temporal and spatial periodic functions are a possible vehicle for the cognitive dimensional elasticity that conserves WM capacity. 9. Supporting roles are proposed of (a) basal ganglia, as a high-capacity cache for traces of recent experience temporarily suspended from active task-relevant processing and (b) of hippocampus as a phase and interval comparator for oscillating signals, whose spatiotemporal dynamics are topologically equivalent to a toroidal grid. PMID- 10800742 TI - Melatonin add-on in manic patients with treatment resistant insomnia. AB - 1. A profound alteration of circadian rhythm of sleep is often a central feature in manic syndrome. Melatonin (MLT) is a main synchronizer of the sleep/wake cycle, playing a role of transduction to brain functioning of informations about periodical environmental changes, i.e. the duration of daylength. 2. In several sleep phase disorders, MLT exerts a therapeutic effect, by normalizing the sleep/wake cycle. 3. Eleven patients, 8 males and 3 females, aged 22-43, meeting DSM IV diagnostic criteria for Bipolar Disorder, Manic Type, were selected for the presence of insomnia not responding to usual hypnotic therapies (benzodiazepine). 4. All the patients were on antimanic treatment. MLT 3 mg per os was administrated at 22.30 h for 1 month, without changing the previous antimanic and hypnotic treatments. All patients showed a significantly longer duration of sleep following MLT add-on. The severity of mania showed a parallel significant decrease. 5. The results of this pilot clinical study suggest that MLT add-on can be useful in antimanic therapy to treat resistant circadian sleep alterations as well as consequently exert a global therapeutic action on the manic state. PMID- 10800743 TI - Increased CBF velocity during word fluency in Huntington's disease patients. AB - 1. This study examined the effects of word fluency and reading on cerebral blood flow in Huntington's disease (HD) patients. 2. Changes in cerebral flow velocity in the anterior (ACA) and middle (MCA) cerebral arteries were measured with functional transcranial Doppler ultrasonography (fTCD) in 13 normal controls and 9 gene positive HD patients. To control for motor effects of word fluency, two "control" conditions, including silent word fluency and a reading test, were also administered to all subjects. 3. Cerebral blood flow velocity was increased during the out loud word fluency test in the ACA, but not MCA, in the HD group compared to controls. This increase was due to motor components of the test, as during silent word fluency the HD group had a decrease in cerebral blood flow relative to controls. Significant correlations between blood flow in the ACA and word fluency test scores were found. Cerebral blood flow velocity during testing also was able to predict group assignment (i.e., control vs. mild HD vs. moderate HD). 4. These findings add to a growing body of literature suggesting that CBF velocity in HD is abnormal during cognitive and motor tasks. Although previous work reported that CBF velocity in HD is decreased during hand use on a maze test, the current experiment finds that speech production increases cerebral blood flow velocity in HD patients. Collectively, these results point to a fundamental disturbance in the regulation of CBF in HD. Mechanisms that could account for these findings, including the potential involvement of nitric oxide, are discussed. PMID- 10800744 TI - Dopaminergic function and the cortisol response to dexamethasone in psychotic depression. AB - 1. It has been hypothesized that psychotic symptoms in depression may be due to increased dopamine activity secondary to hypothalamic-pituitary-adrenal (HPA) axis overactivity. 2. To test this hypothesis, the authors examined the cortisol response to dexamethasone suppression test (DST, 1 mg orally) and multihormonal responses to apomorphine (APO, 0.75 mg s.c.)--a dopamine agonist--in 150 drug free hospitalized patients with DSM-IV major depressive episode with psychotic features (MDEP, n=35), major depressive episode without psychotic features (MDE, n=74), or schizophrenia paranoid type (SCZ, n=41), and 27 hospitalized healthy controls (HCs). 3. MDEPs showed increased activity of the HPA system (i.e. higher post-DST cortisol levels) than HCs, SCZs and MDEs. However, there were no differences in adrenocorticotropic hormone (ACTH), cortisol, prolactin and growth hormone (GH) responses to APO between MDEPs and MDEs and HCs. On the other hand, SCZs showed lower APO-induced ACTH stimulation and a higher rate of blunted GH than HCs, MDEs and MDEPs, suggesting a functional alteration of the hypothalamic dopamine receptors in SCZs. 4. In the total sample and in each diagnostic group, DST suppressors and non-suppressors showed no differences in hormonal responses to APO. 5. These results suggest a lack of causal link between HPA axis hyperactivity and dopamine dysregulation. In contrast to schizophrenia, psychotic symptoms in depression seem not to be related to dopamine function dysregulation. PMID- 10800745 TI - Increased serum arginase activity in depressed patients. AB - 1. Arginase, an important part of the arginine-regulating system modulates nitric oxide generation; a neuroregulatory agent, which has been implicated in various neuropathological conditions. 2. In this regard, the authors investigated the arginine-nitric oxide pathway by measuring serum arginase activity in drug free major (n=18) and minor depressed outpatients (n=12) and healthy control subjects (n=30) in order to make a contribution to the understanding of disease mechanism. 3. Major depressed patients were found to have significantly higher serum arginase activity compared to controls (p<0.001) and minor depressives (p=0.001). Moreover, there was significant positive correlation between arginase activity and severity of depression in patients (p<0.001). 4. Results suggest that the arginine-nitric oxide pathway is involved in depression. Enhanced arginase activity in major depressed patients possibly leading to a decrease in nitric oxide synthesis may contribute to the symptomatology of depression. PMID- 10800746 TI - Effect of adjunctive cortisol on serum sodium in a polydipsic hyponatremic schizophrenic patient. AB - 1. Many polydipsic schizophrenics exhibit enhanced antidiuretic hormone (ADH) activity and thus are hyponatremic and suffer life-threatening water intoxication. Excess cortisol inhibits ADH, while cortisol insufficiency produces impairments in water balance resembling those seen in hyponatremic schizophrenics. Furthermore, hyponatremia normally upregulates cortisol receptors on the neurons which synthesize ADH, which should make them more sensitive to the effects of cortisol. 2. The author treated a hyponatremic schizophrenic, whose water imbalance was unresponsive to standard clinical interventions including clozapine, with a 4-week open trial of 60 mg cortisol daily, followed by a three week taper. 3. Mean serum sodium levels appeared to increase modestly from 114.3 to 118.5 mEq/l while the patient received adjunctive cortisol (P < .06). 4. While a modest effect was seen, the results do not suggest that adjunctive cortisol will reverse hyponatremia, and instead support other data indicating that these patients exhibit a central resistance to glucocorticoid actions. PMID- 10800747 TI - Morphological changes in neuropeptide Y-positive fiber in the hippocampal formation of schizophrenics. AB - 1. The authors observed NPY-positive fibers in the CA4 area of the hippocampus from schizophrenics and normal controls using immunohistochemical techniques. 2. Positive fibers followed a straight course and were oriented to exit the CA4 region of hippocampus in normal controls. 3. Many NPY-positive fibers in the CA4 area appeared coiled or helix-like or appeared wasted and thread-like in schizophrenic brains, compared to those of normal controls. 4. These findings may indicate a dysfunction of the interneuron in the schizophrenic brain and support the hypothesis of developmental impairments of the CNS in schizophrenia, and these morphological changes in fibers may relate to schizophrenic symptoms such as memory or/and learning deterioration. PMID- 10800748 TI - Effects of chronic neuroleptic administration on the hypothalamo-pituitary gonadal axis of male schizophrenics. AB - 1. The authors investigated the effects of long-term neuroleptic administration on the hypothalamo-pituitary-gonadal (HPG) axis of regularly medicated schizophrenic male patients. 2. The subjects were 56 patients who were diagnosed according to the DSM-IV criteria for schizophrenia. Each patient gave informed consent for the research involved in this study. Based on the dosages of neuroleptics, the subjects were classified into two age-matched groups: those with lower dosages (LD) and those with higher dosages (HD). 3. (1) The mean level of the blood testosterone (T) in the LD group was significantly lower than that in the normal group, although the level was within normal range. There was no significant difference in the mean level of the blood T for the HD and the normal groups, or for the HD and the LD groups. (2) There was no significant difference in the mean levels of the blood luteinizing hormone between all groups. (3) Also, there was no significant difference in the mean levels of the blood follicle stimulating hormone between the groups. (4) The mean levels of the blood prolactin in both the LD and HD groups were significantly higher than that in the normal subjects, but there was no significant difference between the LD and HD schizophrenics. 4. Overall, these results seem to indicate that (i) in male patients with chronic schizophrenia, HPG function is impaired, and (ii) chronic neuroleptic medication stimulates the HPG axis in schizophrenic patients with impaired HPG function. PMID- 10800749 TI - Inhibition of nitric oxide synthase in the ventral tegmental area attenuates cocaine sensitization in rats. AB - 1. Male Sprague-Dawley rats were pretreated via bilateral infusion of the VTA with the selective neuronal nitric oxide synthase inhibitor, 7-nitroindazole (0, 8, or 40 ng/hemisphere), prior to each of 7 daily systemic cocaine (30 mg/kg, i.p.) or saline (1 ml/kg) treatments. 2. After a 7-day treatment withdrawal period, rats received a final systemic challenge with either cocaine (30 mg/kg, i.p.) or saline (1 ml/kg). 3. Locomotor and stereotypic activity were measured following the first and last treatments. 4. Daily cocaine treatment led to the development of sensitization to its stereotypic effects as revealed upon drug challenge. 5. The development of sensitization of cocaine-induced stereotypy was completely blocked by daily intra-VTA pretreatment with 7-nitroindazole. 6. In addition, attenuation of the locomotor effects of cocaine challenge was also observed in animals that received daily intra-VTA 7-nitroindazole (40 ng/hemisphere) infusions. 7. The results indicate that VTA nitric oxide is necessary for the development of sensitization of cocaine-induced stereotypic behavior, and that its repeated inhibition may produce lasting effects on the locomotor response to the drug. PMID- 10800750 TI - Effect of R-(-)-alpha-methylhistamine and thioperamide on in vivo release of norepinephrine in the rat hippocampus. AB - 1. The modifications of hippocampal release of norepinephrine following the administration of R-(-)-alpha-methylhistamine and thioperamide, respectively agonist and antagonist of histamine H3 receptors, were assessed in freely moving rats by microdialysis. 2. Both the systemic (2 mg/kg i.p.) and local (100 microM via the probe) administration of thioperamide caused no modifications of basal release, indicating that the histaminergic system is not tonically involved in regulating the hippocampal noradrenergic activity. 3. R-(-)-alpha-methylhistamine (1 and 100 microM) produced a slight, short-lasting and dose-dependent reduction of norepinephrine release antagonized by local perfusion (100 microM) and prevented by systemic administration of thioperamide 2 mg/kg. 4. The results seem to indicate that the modulation of norepinephrine release through presynaptic H3 receptors in the rat hippocampus plays a minor role in the memory-enhancing effects of thioperamide. PMID- 10800751 TI - A low dose of lithium chloride selectively induces Fos protein in the central nucleus of the amygdala of rat brain. AB - 1. Lithium is a very effective treatment for mood disorders. To elucidate the neural substrates of the mood stabilizing actions of lithium, in the present study the authors investigated the effects of a low dose of lithium on regional expression of Fos protein. 2. The administrations of a high dose of lithium chloride (100 mg/kg) induced Fos in widespread areas of the rat brain. In contrast, administration of a low dose of lithium chloride, equivalent to a therapeutic dose in humans, induced Fos only in the central nucleus of the amygdala. 3. These results demonstrate that the central nucleus of the amygdala plays important role in the neural framework that is responsible for the mood stabilizing effect of lithium. PMID- 10800752 TI - Analysis of the role of histamine in inhibitory avoidance in goldfish. AB - 1. Teleost fish have histaminergic cell bodies on the posterior part of the basal hypothalamus. It was suggested that they are homologous to the tuberomammillary E group in rats. However, unlike in rats, fish have fewer ascending fibers. The main projection runs through the ventral telencephalic area reaching the dorsal telencephalon. This projection is considered homologous to the prosencephalic forebrain bundle. 2. The aim of this study was to verify if the histaminergic system has an inhibitory action on learning and memory in goldfish, as suggested previously for higher vertebrates. 3. A two-compartment aquarium with a central sliding door was used. The animals were placed in one of them, the central door was opened after 30 sec and the time spend for crossing between compartments was recorded. After the fish dorsal fin crossed the line between the compartments a 45 g weight was dropped into the compartment the fish entered. 4. On the training day this procedure was done 3 times. Immediately after the 3rd trial the fish was injected i.p. with either vehicle (2 ml/kg), chlorpheniramine (CPA; 1.0, 4.0 and 8.0 mg/kg) or histidine (500 mg/kg). On the next day, fishes were placed in the start compartment and the latency to cross between compartments was again recorded. 5. The group treated with CPA at the dose of 8 mg/kg, presented a significant increase in the latency to leave the start compartment (Wilcoxon rank sum test, p<0.0232). On the other hand, the vehicle and 1-histidine (500 mg/kg) treated groups, presented a decrease in test latency. 6. Thus, we suggest that also in fish, the histaminergic system has an inhibitory role on learning and memory. PMID- 10800753 TI - Behavioral effects of intra-nigral microinjections of manganese chloride: interaction with nitric oxide. AB - 1. Microinjection of manganese chloride (MnCl2) into the rat substantia nigra pars compacta (SNc) induces a neurodegenerative process manifested by apomorphine induced rotational behavior. Manganese intoxication produces a parkinsonism-like phenotype in humans. 2. In addition to motor control the substantia nigra has also been proposed to be related to epilepsy and emotional behavior. 3. Although nitric oxide (NO) participation in neurodegenerative processes is still questioned, neurons stained for NAPDH-diaphorase, a marker of NO-producing cells, are spared in several experimental neuronal lesions. Additionally, NO has also been suggested to participate in motor control. 4. The objective of this study was to analyze the effects of MnCl2-induced nigral degeneration in audiogenic seizure susceptibility, anxiety and motor activity. We also analyzed if NO synthesis inhibition (N(G)-nitro-L-arginine 25 mg/Kg twice a day for 4 days) modifies MnCl2-induced neurodegenerative process. 5. MnCl2 (50 microg) microinjection into the SNc caused a statistical significant higher number of apomorphine (0.75 mg/kg s.c.)-induced rotations. No sensitization to audiogenic seizure was found but the lesion induced an increase of open arm exploration in the elevated plus maze, suggesting an anxiolytic effect. 6. The MnCl2-nigral lesion was accompanied by an increased number of NADPH-d positive neurons in the ipsilateral SNc and striatum (both sides). NO synthesis inhibition potentiated the MnCl2-nigral lesion and reversed the NADPH-d cell number increase. 7. The present results show that MnCl2-nigral lesion can influence emotional behavior and suggest that NO may modify the progression of manganese-induced degenerative process. PMID- 10800754 TI - Effects of some GABA and NMDA antagonists on a model of presynaptic hippocampal paired pulse inhibition. AB - 1. The effects of some NMDA antagonists (7-chlorokynurenic acid and CGS 19755) and of the GABA antagonist penicillin were tested in a model of presynaptic short term paired-pulse inhibition elicited in rat hippocampal slice with high (+ 2 mM) calcium solutions subjected to paired (15 ms)-pulse stimulation paradigm. 2. In control condition a 15 ms paired-pulse stimulation delivered at the level of stratum radiatum, as revealed by the ratio between amplitudes of the conditioned and unconditioned CA1 population spikes (R2/R1), ranging from 1.27 to 2.57, a clear paired-pulse facilitation occurred. Slice perfusion with high (+ 2 mM) calcium shifted, within 30 min, as revealed by a significant (P<0.01) decrease in R2/R1 ratio, paired-pulse facilitation into inhibition. Further perfusion together to high (+ 2 mM) calcium with 0.5 mM penicillin or with 50 microM CGS 19755, but not with 50 microM 7-chlorokynurenic acid significantly decreases the degree of paired-pulse inhibition as revealed by a significative increase in the R2/R1 ratio. 3. The data, demonstrating an inhibitory influence of specific NMDA antagonists in a model a presynaptic paired-pulse inhibition, were discussed in relation with the specific psychodysleptic effects elicited by the drugs in animals and humans. PMID- 10800755 TI - In vivo effects of the putative cognitive enhancer KA-672.HCl in comparison with 8-hydroxy-2-(di-N-propylamino) tetralin and haloperidol on dopamine, 3,4 dihydroxyphenylacetic acid, serotonin and 5-hydroxyindoleacetic acid levels in striatal and cortical brain regions. AB - 1. KA-672.HCl (7-methoxy-6-[3-[4-(2-methoxyphenyl)piperazin-1-yl]propoxy]-3,4-di methyl-2H-1-benzopyran-2-one hydrochloride), designed as a cognitive enhancer, has been investigated through behavioural and binding studies. However, little is known about its biochemical effects on the dopaminergic and serotoninergic system in vivo. 2. In the present study the authors investigated the effects of KA 672.HCl (0.1 mg/kg and 1 mg/kg), 8-hydroxy-2-(di-N-propylamino)tetralin (8-OH DPAT) (1 mg/kg), haloperidol (0.1 mg/kg) and a mixture of haloperidol and 8-OH DPAT on dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT) and 5-hydroxyindolacetic acid (5-HIAA) levels, in striatum and cerebral cortex of rats. 3. Male Wistar rats received an intraperitoneal injection of the drugs or vehicle 1 hour before striatal and cortical brain tissues were dissected out for neurochemical analysis. 4. KA-672.HCl, 8-OH-DPAT and haloperidol significantly reduced striatal DA levels, whereas only KA-672.HCl significantly reduced cortical DA levels. 8-OH-DPAT and haloperidol induced a significant increase in cortical DOPAC levels but only haloperidol significantly elevated the striatal DOPAC content. In contrast, only the higher dose of KA-672.HCl elevated striatal DOPAC levels. Furthermore, KA-672.HCl significantly reduced striatal 5-HT levels and slightly elevated striatal 5-HIAA concentrations. 8-OH-DPAT significantly decreased striatal 5-HIAA levels. All substances were able to enhance the cortical and striatal DA turnover. 5. The cortical and striatal 5-HT turnover was significantly decreased following 8-OH-DPAT treatment and significantly increased in the striatum after haloperidol and KA-672.HCl treatment. 6. The data suggest that KA-672.HCl possesses D2 antagonistic as well as 5-HT1A agonistic properties. However, additional mechanisms of actions by interaction with other neurotransmitter systems such as acetylcholine, excitatory or inhibitory amino acids need to be determined. PMID- 10800756 TI - Switching outpatients between atypical antipsychotics. AB - 1. Some reports have suggested an increase in symptoms when switching patients with psychosis from clozapine to other atypical antipsychotics. 2. No data are available on switching between atypical antipsychotics other than clozapine, though this is common in clinical practice. 3. Six patients with schizophrenia or schizoaffective disorder, bipolar type were switched to quetiapine after finishing a clinical trial of sertindole. 4. During the observation period of two to ten weeks no subjects worsened and one improved. Side effects were mild. 5. These preliminary data suggest that switching between some atypical agents may be well tolerated. Larger controlled trials are needed to confirm this observation. PMID- 10800757 TI - FSH treatment improves sperm function in patients after varicocelectomy. AB - One-hundred and eighty-three patients affected with idiopathic left varicocele, aged between 18 and 45 years, surgically treated have been studied. They were divided in 3 subsets according to sperm count: group A: <10 x 10(6)/ml, group B: 10-20 x 10(6)/ml, group C: >20 x 10(6)/ml. Six months after surgery 115 patients were treated for 3 months with pure human FSH: 75 IU i.m. every other day, while 68 patients treated with placebo served as control group. After therapy, group A showed a significant clear-cut improvement of sperm parameters: count, forward progression, swollen tails and cervical mucus penetration test (CMPT). In group B a significant improvement of sperm motility, viability, DNA integrity and CMPT was observed, while in group C only a significant improvement of CMPT was observed. In conclusion, it can be suggested that FSH treatment in patients after varicocelectomy could improve spermatogenesis, particularly in those who previously have more compromised sperm quality. On the contrary, no significant difference of sperm patterns was recorded in the control group before and after placebo. PMID- 10800758 TI - Iodine status and thyroid volumes of school age children from Northern Cyprus. AB - In an epidemiological study carried out in eight different inland and coastal areas from northern part of Cyprus Island 625 School Age Children (SAC) aged 9 and 10 years, underwent thyroid ultrasonography for determination of thyroid volume (TV). Urinary iodine concentration (UIC) was measured from morning urine samples taken from all of the children. Mean TV was 3.2+/-1.0 ml, 3.3+/-0.8 ml and 3.8+/-1.1 ml, 3.7+/-1.1 ml for 9- and 10-year-old girls and boys respectively. None of the SAC had TV exceeding recommended upper limits for their age and gender. Median UIC of the whole group was 120 microg/l (11-900 microg/l) and only 12.6% of the SAC had UIC below 50 microg/l indicating adequate iodine intake. In this first epidemiological survey conducted in the northern part of the Cyprus Island, iodine status was found to be satisfactory and goiter does not pose a public health problem at the time of the study. Periodical surveys are required especially for the regions which have marginally adequate values and for a coastal region with a significantly higher median value of UIC compared to the others (ie 458 microg/l) (p<0.001). PMID- 10800759 TI - Idiopathic hypercalciuria: O2(-)NO relationship and altered bone metabolism. AB - The pathogenesis of idiopathic hypercalciuria (IH) has not been elucidated yet, but a correlation between IH and altered bone metabolism has been proposed. Since nitric oxide (NO) regulates osteoclasts' bone resorption, a possible role for NO can be suggested. In this study we evaluated iNOS gene expression by reverse transcription of mRNA from monocytes, followed by polymerase chain reaction in patients with IH subdivided into fasting (FH) and absorptive (AH) hypercalciuria. Since superoxide (O2-), which metabolizes NO, is overproduced by osteoclasts during bone resorption, peroxynitrite plasma level was evaluated as index of O2-. Vertebral BMD in IH as a whole group was lower vs controls (C) (Z score=-1.78+/ 0.2 vs 0.51+/-0.25, p<0.001), but only FH patients showed a reduced bone density (2.13+/-0.18 vs 0.51+/-0.25, p<0.0001). PTH and calcitriol were not different. FH showed an increase in b-ALP vs AH and C (41.1+/-2.6 vs 30.1+/-3.9 vs 26.6+/-3.6 U/l p<0.02), and higher uHP, either on NCD (17.7+/-1.6 vs 11.4+/-1.3 mg/g uCr, p<0.04) or after LCD (26.7+/-2.5 vs 16.7+/-1.9, p<0.01). Cells from FH patients, but not from both AH patients and C, expressed iNOS. Peroxynitrite plasma level was elevated in FH (0.30+/-0.07) pmol/l while not detectable in AH and C. This study confirms an altered bone metabolism only in FH which shows an abnormal NO system. The increased iNOS gene expression in FH, in fact, points toward an altered NO system's activity downstream the generation of NO. A possible interaction of NO with O2-, which breaks down NO, and the role of this interaction in the pathophysiology of IH is discussed. PMID- 10800760 TI - GH/IGF-I axis in Prader-Willi syndrome: evaluation of IGF-I levels and of the somatotroph responsiveness to various provocative stimuli. Genetic Obesity Study Group of Italian Society of Pediatric Endocrinology and Diabetology. AB - Basal IGF-I levels and the GH response to at least two among provocative stimuli such as clonidine (CLO, Catapresan, 150 mcg/m2 p.o.), GHRH (1 mcg/kg i.v.)+arginine (ARG, 0.5 g/kg i.v. infusion during 30 min) and GHRH+pyridostigmine (PD, Mestinon cpr 60 mg p.o.) have been evaluated in 43 children with Prader-Willi syndrome (PWS, 17 males and 26 females, age 3-22 yr, 7 normal weight and 36 obese PWS), in 25 normal short children (NC, 17 males and 8 females, 7.7-18.5 yr) and in 24 children with simple obesity (OB, 14 males, 10 females, 7.7-21.5 yr). Both normal weight and obese PWS had mean IGF-I levels lower than those recorded in NC (p<0.001) and OB (p<0.001). The GH responses to GHRH+ARG and GHRH+PD in NC were similar and higher than that to CLO (p<0.001). In PWS the GH response to GHRH+ARG was higher than that to GHRH+PD (p<0.001) which, in turn, was higher than that to CLO (p<0.001); these responses in PWS were lower than those in normal children (p<0.02) and similar to those in OB. In normal weight PWS the GH responses to GHRH+ARG and to GHRH+PD were similar and higher than to CLO (p<0.05); however, each provocative stimulus elicited a GH rise lower than that in NC (p<0.05). In obese PWS as well as in OB the GH response to GHRH+ARG was higher than that to GHRH+PD (p<0.02) which, in turn, was higher than that to CLO (p<0.001); all GH responses in obese PWS and OB were lower than those in NC (p<0.001) but similar to those in normal weight PWS. In conclusion, patients with PWS show clear reduction of IGF-I levels as well as of the somatotroph responsiveness to provocative stimuli independently of body weight excess. These results strengthen the hypothesis that PWS syndrome is frequently connotated by GH insufficiency. PMID- 10800761 TI - GnRH receptors in human breast cancer and its contiguous not-involved breast tissue. AB - The present study was undertaken to evaluate the presence of GnRH receptors (GnRH R) in breast cancer and not-involved breast tissue, and the relationships between GnRH-R and receptors for estrogen (ER) and progesterone (PgR) in the same tissues. Utilizing a tritiated natural GnRH in order to assay the native receptor binding we analyzed the level of binding sites for GnRH in membranes derived from 90 breast tumors and in 40 cases from neighboring, not-involved breast tissue. GnRH-R was found both in cancer and normal tissues. The prevalence for GnRH-R was higher in tumor than in not-tumor tissue (45% vs 39%, respectively), but the overall levels were not significantly different (15.9+/-24 fmol/mg protein vs 18.2+/-39 fmol/mg protein, respectively). The only statistically different content of GnRH-R we found concerned PgR negative vs PgR positive tumor tissues (mean content: 23 vs 11 fmol/mg protein, respectively in PgR- and PgR+ tumors, p=0.03 by t test); furthermore the proportion of GnRH-R positive cases in the tumor resulted significantly higher in premenopausal patients vs postmenopausal (56% vs 32%, by Chi square test, p<0.05). The GnRH receptors status of primary tumor and contiguous not-involved breast tissue resulted associated (overall agreement: 63%, p<0.05) but no specific steroid patterns for GnRH-R positivity was observed. PMID- 10800762 TI - Sequential administration of arginine and arginine plus GHRH to test somatotroph function in short children. AB - The hormonal diagnosis of GH deficiency in childhood is conventionally based on the GH response to at least two provocative stimuli. Among these, arginine (ARG) has long been considered a classical, centrally mediated stimulus of GH secretion. ARG is also able to potentiate the GH response to GHRH, likely inhibiting hypothalamic somatostatin; this combined test is one of the most potent to explore the maximal secretory capacity of somatotroph cells. Based on these premises, we verified whether the sequential administration of ARG and ARG+GHRH could be feasible as single step provocative test to evaluate the GH releasable pool in short children. To this goal, 48 normal short children (35 M and 13 F, 12.0+/-0.4 yr, PS 1: 255 II-IV: 23) underwent a test with ARG (0.5 g/kg i.v. from 0 to +30 min) followed by a coadministration of ARG (from +120 to 150 min) plus GHRH (1 microg/kg i.v. at +120 min). ARG alone elicited a clear GH response (mean peak vs baseline: 12.1+/-1.7 vs 2.0+/-0.4 microg/l, p<0.001, Cmax range 12-51.0 microg/l). Following this GH rise, the hormonal levels at +120 min approached to baseline levels (4.2+/-0.8 microg/l) but then showed marked response to the coadministration of ARG+GHRH. The GH peak following ARG+GHRH (mean peak: 47.8+/-3.3 microg/l, p<0.001; Cmax 22.4-150.0 microg/l) was clearly higher (p<0.001) than that recorded after ARG alone. The GH responses to both ARG and ARG+GHRH were independent of gender, puberty, height velocity, body mass index (BMI) and IGF-I levels. Nine normal short children (16%) had GH peaks lower than 7 microg/l after ARG alone, while none showed GH peak below 20 microg/l after ARG+GHRH. Thus, ARG alone is a good stimulus of GH secretion but false positive responses frequently occur in normal short children. ARG+GHRH is a more potent stimulus giving no false positive responses even after previous challenge with ARG alone. Testing with sequential administration of ARG and ARG+GHRH may allow the single step evaluation of the somatotroph response to central and pituitary stimuli in short children. PMID- 10800763 TI - High prevalence of thyroid abnormalities in a Chilean psychiatric outpatient population. AB - The aim of the present study was to establish the prevalence of thyroid disturbances in patients consulting for panic and mood disorders. These data may be relevant because thyroid functional alterations affect the success of treatment in these pathologies. We studied prospectively 268 psychiatric outpatients (204 females and 64 males) diagnosed by DSM-IV criteria. We excluded patients with addictive disorders and major medical disease. We measured TSH, Free T4 (FT4) and antimicrosomal antibodies (AMA). We diagnosed classical hypothyroidism when the TSH value was >10 microUI/ml (NV=0.25-4.3) and subclinical hypothyroidism when the TSH value was between 5-10 microUI/ml. Hyperthyroidism was diagnosed when FT4 >1.4 (NV=0.8-1.4), the TSH suppressed and the radioiodine uptake >20% (NV=5-15). Positive antimicrosomal antibodies (AMA) titres were >1:100 dilution. Hypothyroidism was diagnosed in 26/268 patients (9.7%); 10 cases corresponded to the classical form (38.5%) and 16 cases to the subclinical form (61.5%). Hyperthyroidism was found in 6/268 patients (2.2%). Normal thyroid function with positive AMA was found in 28/268 patients (10.4%). Hypothyroidism was more common in patients with mood disorders, and hyperthyroidism in patients with panic disorders. Patients with panic disorder had significant higher levels of FT4. The prevalence of positive AMA, hypothyroidism and hyperthyroidism was higher in women than men. We found a high frequency of thyroid abnormalities in a psychiatric outpatient population. These data suggests that routine evaluation of thyroid function should be considered in patients consulting for mood and panic disorders. PMID- 10800764 TI - From macroprolactinoma to concomitant ACTH-PRL hypersecretion with Cushing's disease. AB - Multiple pituitary hormone hypersecretions have been already described, but the combination of PRL and ACTH excess is rare. This report deals with a 42-yr-old woman affected by macroprolactinoma (PRL 12,720 microg/l, huge tumor with extrasellar extension at imaging). After one year on dopaminergic treatment causing PRL normalization and tumor shrinkage, she developed hypercortisolism (UFC 1,000 microg/24 h, ACTH 200 ng/l). Cushing's disease was diagnosed. After neurosurgery (at immunocytochemistry mixed ACTH-PRL adenoma was shown) hypercortisolism remitted, whereas pathological hyperprolactinemia with tumor remnant in cavernous sinus persisted and hypopituitarism developed. The patient reported seems atypical for the following reasons: 1) the concomitant PRL and ACTH hypersecretions; 2) the clinical presentation with hypercortisolism following hyperprolactinemia; 3) the surgical cure of hypercortisolism with persisting hyperprolactinemia. PMID- 10800765 TI - Adrenocortical carcinoma manifesting pure primary aldosteronism: a case report and analysis of steroidogenic enzymes. AB - Adrenocortical carcinoma manifesting pure hyperaldosteronism is extremely rare. We report here a 61-year-old woman with biochemically proven primary aldosteronism due to right adrenocortical carcinoma. Computed tomographic scan showed 4.5x5.3 cm lobulated mass with tiny calcification, while there was no significant uptake of 131I-iodomethyl norcholesterol in the tumor. Immunohistochemical analysis demonstrated expression of steroidogenic enzymes in the tumor tissue: P-450scc, P-45c21, 3beta-hydroxysteroid dehydrogenase, P450(17alpha), and P-450(11beta). In addition, we could demonstrate mRNA expression of aldosterone synthase (P-450aldo:CYP11B2) in the tumor by specific ribonuclease protection assay. This is the first report of a case of primary aldosteronism due to adrenocortical carcinoma, in which expression of all sets of steroidogenic enzymes required for aldosterone synthesis was proven. PMID- 10800766 TI - Gonadotropin-releasing hormone-induced partial empty sella clinically mimicking pituitary apoplexy in a woman with a suspected non-secreting macroadenoma. AB - Pituitary apoplexy has been reported as a rare complication of dynamic testing used for the study of pituitary functional reserve. In 1993, a diagnosis of non secreting macroadenoma with moderate functional hyperprolactinaemia was made in a 43-year-old woman. Soon after the start of therapy with bromocriptine up to 5 mg/die, the patient complained of nausea and postural hypotension. As the symptoms persisted even when the dose was reduced to 2.5 mg/die, the patient was transferred to therapy with quinagolide at the dosage of 37.5 microg/die. PRL levels quickly normalized (range 1.4-5.7 ng/ml) as well as menstrual cycles, and no side-effect was reported. In 1995 a sellar magnetic resonance imaging (MRI) showed no shrinkage of the known macroadenoma. In 1996, few hours after a gonadotropin-releasing-hormone (GnRH) test, which showed normal LH and FSH response and with baseline PRL levels in the normal range, the patient started complaining of severe frontal headache, nausea and vomiting. No gross visual defects were present. An emergency computed tomography (CT) showed no evident hemorrhagic infarction in the macroadenoma. The symptoms completely resolved in few days with steroidal and antiemetic therapy. A new MRI performed in 1998 showed a partial empty sella and PRL levels were in the normal range under dopaminergic treatment. The pituitary functional reserve proved normal on dynamic testing. The temporal association between the onset of symptoms and the GnRH test strongly suggests an association between the two events. No evident signs of pituitary apoplexy (either on emergency CT or hormonal evaluation) were detected. The authors suggest that GnRH can cause severe side-effects that mimic pituitary apoplexy without related morphological evidence and that, in our particular case, it can have caused the gradual disappearance of the non-secreting macroadenoma. Moreover, a causal role of the chronic dopaminergic treatment cannot be completely ruled out. PMID- 10800767 TI - Microprolactinoma: medical or surgical treatment as first line approach? The case for medical therapy. PMID- 10800769 TI - Cabergoline, prolactin and melatonin release at night in healthy men. PMID- 10800768 TI - Genetic aspects of central hypothyroidism. AB - Central hypothyroidism, characterized by insufficient TSH secretion in the presence of low levels of thyroid hormones, is a rare disorder. It has recently been found that, although mainly due to tumors or infiltrative diseases of the hypothalamo-pituitary area or to pituitary atrophy, central hypothyroidism may be caused by inactivating mutations in several of the genes that code for the various proteins involved in the regulation of the hypothalamo-pituitary-thyroid axis (HPTA). These experiments of nature allow us to better understand the pathophysiology but also the normal physiology of the HPTA. This review will analyze reports of mutations that affect the HPTA and result in either isolated central hypothyroidism or in the syndrome of combined pituitary hormone deficiency (CPHD). Mutations have been identified in the genes for the TRH receptor, the transcription factors Pit-1 and PROP1, and the TSH beta-subunit. PMID- 10800770 TI - "A gentleman". Giovanni Antonio Amadeo. Pavia 1447-Milan 1522. PMID- 10800771 TI - Thyroid hormone increases the conductance density of f-channels in rabbit sino atrial node cells. AB - In hyperthyroidism, the main cardiac manifestation is an increase in resting heart rate with a great degree of sinus tachycardia. As the pacemaker current, I(f), is reputed to be the main ionic mechanism involved in sinus rhythm, the objective of this study was to investigate the effect of triiodothyronine (T3) on I(f), in sino-atrial (SA) node cells isolated from rabbit heart, using the patch clamp technique in whole-cell configuration. In cells incubated for 5-7 h in a medium containing 1 microM T3, the hormone increased I(f) by rising the maximal conductance without inducing shift of the activation curve or change in the reversal potential. The mean values of I(f) density (42.05+/-5.93 pA/pF, n = 13) and maximal conductance density (854.25+/-124.83 pS/pF, n = 16) were significantly increased compared with those measured in untreated cells (20.21+/ 2.53 pA/pF, n = 14; 392.69+/-54.95; pS/pF, n = 16). In cells treated with T3, 0.1 microM isoprenaline, a beta-adrenergic agonist, was still able to induce a positive shift of the voltage dependence of I(f) activation. Reverse T3, the inactive form of T3, used in the same conditions, had no effect on I(f). According to the results, it was suggested that the acceleration of resting heart rate observed in hyperthyroidism, might be the consequence of a T3-induced stimulation of f-channel transcription in SA node myocytes. PMID- 10800772 TI - Phosphorylation of the 5-hydroxytryptamine3 (5-HT3) receptor expressed in HEK293 cells. AB - The 5-hydroxytrptamine3 (5-HT3) receptor is a pentameric complex belonging to the family of ligand-gated ion channels. A variety of studies have suggested that phosphorylation regulates the rate of desensitisation and the size of amplitude of the receptor current. In this study we have examined the phosphorylation of the myc-tagged wild-type 5-HT3A receptor subunits from guinea-pig expressed in HEK293 cells (human embryonic kidney). Stably transfected cells were metabolically labelled with 32P-phosphoric acid. The results of immunoprecipitation and autoradiography demonstrate that both splicc variants of the 5-HT3A receptor subunit are phosphorylated in HEK293 cells. Site-specific mutagenesis revealed that phosphorylation occurs at serine 409, a potential target of protein kinase A. Thus the 5-HT3 receptor might be modulated by intracellular pathways, that allow variable 5-hydroxytryptamine action as responses to different extracellular stimuli. PMID- 10800773 TI - Beta3 receptors mediate relaxation in stomach fundus whereas a fourth beta receptor mediates tachycardia in atria from transgenic beta3 receptor knockout mice. AB - Pharmacological studies have revealed a non-beta1, beta2 or beta3 adrenergic receptor that mediates tachycardia in rat and human atria. The present studies utilized transgenic mice that lack the rodent beta3 receptor to explore, in a more definitive fashion, whether a non-beta1, beta2 or beta3 receptor can mediate atrial tachycardia. Insofar as the rat stomach fundus possesses a beta3 receptor mediating relaxation, we examined the stomach fundus from beta3 receptor knockout mice for the presence or absence of the beta3 relaxant receptor. Contractile responses to carbamylcholine were similar in potency and magnitude between mouse stomach fundus from wild type and beta3 receptor knockout animals. However, the classical beta3 receptor agonist CL316243, (10(-8)-10(-6)M) relaxed stomach fundus from wild type mice, but not from the beta3 receptor knockout animals. These data provide functional evidence for the absence of the beta3 receptor in beta3 receptor knockout animals and support the role of beta3 receptors mediating relaxation in mouse stomach fundus. Atria from mice lacking the beta3 receptor responded similarly (in potency and maximal increase in heart rate) to isoproterenol (10(-9)-10(-6)M) as atria from wild type mice. Furthermore, propranolol (3 x 10(-7) M) produced a dextral shift in the concentration response to isoproterenol in atria from both the beta3 receptor knockout and wild type mice with negative log K(B) values of 8.03 and 8.09, respectively. Thus, beta receptors mediating tachycardia to isoproterenol are intact and respond similarly in atria from both knockout and wild type mice. Furthermore, CGP12177, a prototypic 'atypical' beta receptor agonist produced tachycardia with a similar EC50 and maximal response in atria from both the wild type and beta3 receptor knockout mice. Cyanopindolol was a partial agonist relative to CGP12177 in both wild type and beta3 receptor knockout mice. Tachycardia to CGP12177 and cyanopindolol was not blocked by propranolol (3 x 10(-7) M) in atria from either group. These data provide definitive evidence that the receptor mediating tachycardia to CGP12177 and to cyanopindolol in atria from the transgenic beta3 receptor knockout mice is neither the beta1, beta2, nor beta3 adrenergic receptor. PMID- 10800774 TI - Extracellular Ca2+ sensitivity of mGluR1alpha associated with persistent glutamate response in transfected CHO cells. AB - We previously reported that the metabotropic glutamate receptor 1alpha (mGluR1alpha) can be activated not only by glutamate but also by extracellular Ca2+ (Ca2+o), and that Ser 166 in the extracellular domain determines the sensitivity to Ca2+o. In the present study, we investigated by intracellular Ca2+ (Ca2+i) imaging, the effect of Ca2+o on the glutamate responses of Chinese Hamster Ovary (CHO) cells stably expressing mGluR1alpha wild-type (CHO-wt). As a negative control, we carried out similar experiments using CHO cells expressing Ser166Asp mutant of mGluR1alpha (CHO-S166D) or the substance P receptor (CHO SPR), which were not activated by Ca2+o application. We observed a remarkable prolongation of the duration of the glutamate response in CHO-wt cells in a Ca2+o concentration dependent manner. In CHO-S166D cells and CHO-SPR cells, only a small sustained component of the glutamate response was observed in the presence of Ca2+o. These sustained components were blocked by SKF-96365, a blocker of receptor-operated Ca2+-influx. Thus, it was concluded that the Ca2+o-sensing function of mGluR1alpha-wt induced the persistent opening of the receptor operated Ca2+-permeable channels, probably by persistent activation of the receptor by glutamate. We additionally observed that the dose-response relationship of CHO-S166D and CHO-SPR shifted significantly by changing Ca2+o concentration, i.e. Ca2+o was required to maintain the normal ligand responses of these receptors. PMID- 10800775 TI - Different populations of inositol 1,4,5-trisphosphate receptors expressed in the bovine adrenal cortex. AB - The inositol 1,4,5-trisphosphate (InsP3) receptor forms a tetrameric channel responsible for the release of Ca2+ from intracellular stores. In the present study we showed that the experimental approach used to separate bound and free ligands may discriminate between two populations of InsP3 binding sites in bovine adrenal cortex microsomes. A large population of low affinity sites and a small population of high affinity sites were detected with centrifugation and filtration approaches, respectively. Both populations were found in the supernatant and the cytoskeleton fractions of Triton X-100 solubilized microsomes. After treatment of microsomes with thimerosal, an alkylating reagent known to increase InsP3 receptor affinity, the filtration and the centrifugation approaches yielded identical results. With selective anti-InsP3 receptor antibodies, we showed that types 1, 2 and 3 InsP3 receptors are present in intact microsomes and in the cytoskeleton fraction. Binding studies on immunoprecipitated receptors revealed that anti-type 1 antibody recognizes a large population of low affinity sites whereas anti-type 2 antibody recognizes a small population of high affinity sites. Our results indicate that the three types of InsP3 receptors are expressed at different levels in the bovine adrenal cortex. The presence of different types of InsP3 receptors with different ligand binding affinities and their association with the cytoskeleton offer a convenient way for the cell to simultaneously regulate its intracellular Ca2+ concentration and reorganize the spatial distribution of its Ca2+ stores. PMID- 10800776 TI - Patterns of shaker family gene expression in single identified neurons of the American lobster, Homarus americanus. AB - The patterns of expression of voltage gated potassium channel genes of the Shaker family have been mapped in identified neurons of the lobster (Homarus americanus) ventral nerve cord using a single cell reverse transcriptase polymerase chain reaction procedure. Using specific oligonucleotides derived from the sequences of the shaker, shab, and shaw genes of the spiny lobster, Panulirus interruptus, we detected the corresponding potassium channel DNA fragments from Homarus americanus. The Homarus DNA fragments are 87-98% identical at the nucleotide level to the Panulirus DNA fragments. We used the Panulirus primers to measure the complement of RNAs for shaker, shab, and shaw in single identified cells that use GABA, glutamate, octopamine or serotonin as chemical messengers. Shaker and shaw RNAs were found in all four identified neuron types but shab RNA was not detected in serotonin cells under the present experimental conditions. All cells expressed alpha-tubulin RNA, which serves as an internal control suggesting that cells are intact after dissection. In glial cells that surround the neuronal cell bodies, the potassium channel genes are expressed at low to non-detectable levels. PMID- 10800777 TI - Structural elements determining activation kinetics in Kv2.1. AB - Voltage-dependent K+ channels open when depolarizing the membrane voltage. Among the different alpha-subunits, the time course of current activation spreads over a wide range. The structural basis underlying this diversity is not known. We constructed multiple chimeras between two voltage-dependent K+ channels, the rapidly activating Kv1.2 and the slowly activating Kv2.1, and we focused on the C terminal half of the core region. The general strategy was to substitute parts of Kv2.1 by corresponding parts of Kv1.2 and to test for an acceleration of activation. We identified three regions which contribute to the determination of the activation kinetics: the S5-pore linker, the deep pore, and the S4-segment. PMID- 10800778 TI - Does cerebral microembolization during cardiopulmonary bypass impair cerebral autoregulation. PMID- 10800779 TI - Latissimus dorsi reverse flap to substitute the diaphragm after extrapleural pneumonectomy. AB - BACKGROUND: The standard procedure for diaphragm reconstruction after extrapleural pneumonectomy for a malignancy consists of the use of prosthetic patches. Our original technique utilizing the reverse flap of the latissimus dorsi is evaluated. METHODS: Once the extrapleural pneumonectomy is performed, the distal portion of the latissimus dorsi, which has been divided with a standard posterolateral thoracotomy at the level of the fifth to sixth rib, is elevated into the chest through the passage obtained by resection of the tenth rib and sutured to the lower pericardium and to the chest wall. Nine patients were evaluated. RESULTS: No operative death occurred. No flap-related complication nor infection was postoperatively assessed. Six patients received adjuvant radiotherapy. No late complication was observed. CONCLUSIONS: The distal latissimus dorsi can be used for total reconstruction of one hemidiaphragm, ensuring a watertight separation between the pleural and peritoneal cavities and avoiding paradoxical respiratory motion. In our opinion, the technique was easier, faster, and more reliable than the standard procedure employing prosthetic materials. We recommend that the procedure be integrated with the standard technique of extrapleural pneumonectomy. PMID- 10800780 TI - Pharyngeal dysphagia in postesophagectomy patients: correlation with deglutitive biomechanics. AB - BACKGROUND: Because of the transient nature of pharyngeal phase dysphagia, posttranshiatal esophagectomy patients provide a model for studying the correlation of dysphagic symptoms and aspiration with deglutitive biomechanics. METHODS: We studied 8 transhiatal esophagectomy patients (age range, 51 to 78 years) and 8 normal age-matched controls in upright position using lateral and anteroposterior (AP) projection videofluoroscopy during three 5 mL barium swallows. RESULTS: The maximum upper esophageal sphincter (UES) AP diameter and maximum anterior excursion of the hyoid bone in patients with transhiatal esophagectomy who experienced aspiration (6.2+/-0.6 and 9.0+/-2.0 mm, respectively) were significantly smaller than those of age-matched normal controls (9.4+/-0.7 and 17.0+/-1.0 mm, respectively). Resolution of aspiration was associated with a significant increase in AP diameter of the UES as well as anterior and superior excursion of the hyoid bone (p<0.05). CONCLUSIONS: Dysphagic symptoms and aspiration in posttranshiatal esophagectomy patients are associated with significant abnormalities of deglutitive biomechanics. Improvement in deglutitive biomechanics is associated with resolution of dysphagic symptoms as well as postdeglutitive aspiration in these patients. PMID- 10800781 TI - Primary non-Hodgkin's lymphoma of the lung. AB - BACKGROUND: Primary non-Hodgkin's lymphoma of the lung is a rare entity. Although the prognosis is favorable, clinical features, prognostic factors, and patient management have not been clearly defined. METHODS: We reviewed retrospectively the records of 48 patients operated on for primary pulmonary non-Hodgkin's lymphoma. The study group consisted of 21 male (44%) and 27 female (56%) patients with a mean age of 61.8 years. Thirty-seven and a half percent of patients were asymptomatic, and 62.5% were seen with pulmonary symptoms, systemic symptoms, or both. A definitive diagnosis was obtained by thoracotomy in 90% of patients, thoracoscopy in 8%, and anterior mediastinotomy in 2%. RESULTS: Complete surgical resection was possible in 19 patients (40%). A mucosa-associated lymphoid tissue lymphoma (MALT) was found in 35 patients and lymphoma that was not of this type, in 13. The 1-year, 5-year, and 10-year survival rates were 91%, 68%, and 53%, respectively in the group with mucosa-associated lymphoid tissue lymphoma and 85%, 65%, and 64% in the group with lymphoma that was not of the mucosa associated lymphoid tissue type. None of the prognostic factors studied (mode of presentation, smoking history, bilateral disease, postoperative stage, complete resection, adjuvant chemotherapy, histology) significantly influenced patient survival. CONCLUSIONS: Primary non-Hodgkin's lymphoma of the lung occurs with nonspecific clinical features. Although patient survival is good, prognostic factors could not be identified. PMID- 10800782 TI - A new portable chest drainage device. AB - BACKGROUND: Persistent air leak is a frequent complication in lung operation. The Heimlich valve is the standard system for venting the pleural cavity. The device achieves good results and is well tolerated, but the main problem is when air leak is associated with fluid leakage. METHODS: In order to improve the outpatient management of persistent air and fluid drainage after resectional procedures, we developed an original device. It is a portable system provided with a one-way valve connected to the chest tube for drainage of air and fluid, a reservoir for collecting fluid, and a one-way exhaust valve to evacuate air from the bag. RESULTS: We analyze the advantages of our device versus the Heimlich valve in the first series of 18 selected patients. Our system is drier and cleaner, easier to manage, and ambulatory visits are seldom needed. There is also a cost savings. CONCLUSIONS: Our device enhances ambulation, independence, and the quality of life of the patients, and decreases the need for hospital and outpatient care. PMID- 10800783 TI - Lobectomy for destroyed lung in quadriplegic patients. AB - BACKGROUND: Sixty-seven percent of quadriplegic patients after spinal cord injury (SCI) develop respiratory complications, which leads to death in one third. Preventive measures may fail to avoid parenchymal destruction and possible septic complications. METHODS: Three quadriplegic patients (C3-C6 level), with destroyed lower lobes and incontrollable septic symptoms, were subjected to lobectomy. RESULTS: Neither operative morbidity nor mortality was observed. All patients were discharged home without ventilatory assistance, and were symptom-free. CONCLUSIONS: When the endobronchial chronic infection calls for repeated fiberoptic bronchoscopies to clear the bronchial tree, the parenchymal destruction is limited to one lobe of the lung, and there is evidence of impending septic complications, lobectomy may be indicated in quadriplegics to eradicate the source of infection. PMID- 10800784 TI - Palliation of advanced esophageal carcinoma by photodynamic therapy and irradiation. AB - BACKGROUND: We wanted to determine the role of photodynamic therapy in a multimodal approach for the treatment of patients with advanced cancer of the esophagus. METHODS: We reviewed the cases of 119 patients with nonresectable esophageal carcinoma who underwent endoluminal palliation. Twenty-one patients required initial dilation and tumor obliteration with a neodymium: yttrium aluminum-garnet laser prior to therapy. Forty-four patients received photodynamic therapy followed by brachyradiotherapy, and 75 patients were treated with brachyradiotherapy. In both groups, some patients also received external-beam irradiation. RESULTS: Photodynamic therapy produced a significant difference in relieving stenosis caused by tumor stenosis (mean, 6.6 mm; p = 0.0000). The dysphagia score improved by one to three levels in all patients, with a significant difference in favor of PDT (p = 0.0003). The mean number of overall treatment sessions was four (range, one to seven). The rate of major complications was 9.2%. Four esophageal perforations occurred, three after intervention and one spontaneously 5 months later. Four esophagorespiratory tract fistulas developed several months after combined PDT and irradiation. The mean overall survival was 7.7 months, and analysis of variance revealed a significant difference in favor of PDT and external-beam irradiation (p = 0.0129 and p = 0.0001, respectively). CONCLUSIONS: Photodynamic therapy has been shown to be an effective palliative treatment of advanced esophageal cancer. However, proper patient selection is necessary to prevent serious complications. PMID- 10800785 TI - An improved orthotopic xenotransplant procedure for human lung cancer in SCID bg mice. AB - BACKGROUND: Overall prognosis in human lung cancer is still poor. A highly reproducible, easy to perform in vivo model, which closely resembles the clinical features of advanced human lung cancer, is required for the evaluation of novel therapies. METHODS: Tumor cells, originated from a human adenocarcinoma, a squamous cell carcinoma, and an undifferentiated large cell carcinoma, were xenotransplanted heterotopically by subcutaneous and intravenous injection and compared with orthotopic intrapleural and intrapulmonary xenotransplantation by a facilitated engraftment procedure into SCID bg mice. RESULTS: Subcutaneous injection of tumor cells resulted in a 100% engraftment rate with establishment of solid tumors without clinically relevant metastases. Intravenous injection had poor engraftment rates by hematogenous spread. Depending on the cell line, a 80% to 100% engraftment rate in orthotopic xenotransplantation was achieved, resulting in a consistent pattern of mediastinal and bilateral pulmonary metastases. CONCLUSIONS: The facilitated orthotopic xenotransplantation of human lung cancer is easy to perform and results in a reproducible in vivo model that closely resembles the clinical features of advanced human lung cancer. Consequently, this model appears suitable for in vivo evaluation of novel cancer therapies in preclinical tests. PMID- 10800786 TI - The lingula is an appropriate site for lung biopsy. AB - BACKGROUND: Lung biopsy is commonly performed for diagnosis of diffuse pulmonary disease. The lingula offers technical advantages for biopsy, however the quality of tissue obtained by lingula biopsy has been questioned. We sought to determine whether lingula biopsy was a satisfactory site for biopsy in terms of diagnostic yield, therapeutic interventions, and survival results. METHODS: All diagnostic lung biopsies performed for diffuse lung disease at 3 university affiliated hospitals between July 1, 1992 and December 31, 1998 were retrospectively reviewed. Patients were divided into 2 groups, depending upon site of biopsy: patients with lingula biopsy only and those with biopsies from other sites. RESULTS: There were 75 patients; 20 underwent biopsy of the lingula alone, 48 had biopsy of other sites with or without biopsy of the lingula, and location of biopsy was unknown in 7 patients. Histologic diagnosis was achieved in all patients. Significant beneficial therapeutic changes were made in 14 lingula patients, and consisted of immunosuppression in 12 cases. Three patients died in the hospital or within 30 days. Fourteen patients survived 1 year. There was no significant difference between patients that had biopsy of the lingula alone and those that had biopsies from other sites in urgency, technique, histologic diagnosis, rate of therapeutic interventions, hospital mortality, or 1 year survival. CONCLUSIONS: Lung biopsy of the lingula compared to other anatomic sites has equivalent diagnostic yield, therapeutic significance, and survival. Given the technical ease of biopsy, when disease is present radiographically it is the preferred site for lung biopsy. PMID- 10800787 TI - Segmental nonanastomotic bronchial stenosis after lung transplantation. AB - BACKGROUND: Nonanastomotic distal bronchial stenosis has been observed in some patients after lung transplantation. We investigated its relationship with acute cellular rejection (ACR), infection, and ischemia. METHODS: Between January 1994 and December 1997, 246 lung transplantations were performed at our hospital. These cases were retrospectively reviewed and evaluated to identify those patients with nonanastomotic bronchial stenosis. RESULTS: Six patients had bronchial stenosis within the grafted airway distal to the uninvolved anastomotic site. The average ACR before stenosis was 1.9 compared with 1.6 in a control group. ACR at the time of first recognition of the stenosis ranged from A2 to A3.5, with an average value of A2.9. All 6 patients demonstrated alloreactive airway inflammation before and at the time of stenosis. Four patients had evidence of ischemic damage in the perioperative period. CONCLUSIONS: Segmental nonanastomotic large airway stenosis after lung transplantation should be assessed separately from anastomotic complications. Although the pathogenesis is unclear, certainly one should consider alloreactive injury, ischemic damage, and infection as individual and coercive causes. PMID- 10800788 TI - Clinicopathological and biological assessment of lung cancers with pleural dissemination. AB - BACKGROUND: This study provides the surgical outcome of lung cancer patients with pleural dissemination, with the assessment of the clinicopathological and biological prognostic factors. METHODS: Forty-three patients who underwent operations were studied. Vascular endothelial growth factor (VEGF) and autocrine motility factor receptor (AMFR/gp78) expression was immunohistochemically evaluated. RESULTS: In total, the overall 3 and 5-year survival rates were 31.4% and 13.1%, respectively. The patients who underwent the pleuropneumonectomy had a worse outcome than those who underwent limited operations (pleurectomy plus parenchymal resections were less than pneumonectomy). VEGF and AMFR/gp78 were highly expressed in primary tumors. Among the patients who underwent limited operations, pathological types other than adenocarcinoma and high expression of VEGF were significantly associated with a worse outcome. The pathological type was the only characteristic to retain a significant independent prognostic impact on overall survival. CONCLUSIONS: The results imply the validation of limited operation for lung cancer with pleural dissemination for the local control. High frequency of VEGF and AMFR/gp78 expression conform to the interpretation that patients with pleural dissemination have a high-risk of systemic disease. PMID- 10800789 TI - Long-term safety and tolerance of silicone and self-expandable airway stents: an experimental study. AB - BACKGROUND: A variety of respiratory stents are currently available, but the ideal airway prosthesis seems far from being recognized. The objective of this study was to verify safety and long-term effect on the bronchial wall of three different types of airway stents. METHODS: Twelve healthy adult sheep were divided in three groups, scheduled to receive: (1) bare self-expandable metallic stents (Gianturco); (2) silicone stents (Dumon); and (3) covered self-expandable synthetic stents (Polyflex). Insertions were performed through a rigid bronchoscope under general anesthesia. Chest roentgenogram was performed 1 and 6 months after surgery, and flexible bronchoscopy after 6 months. Twelve months postoperatively, the animals were killed and a postmortem examination was carried out. RESULTS: All Polyflex stents migrated during the observation period; one late migration was observed in the Dumon group. Microscopic study showed: (1) Gianturco stents: full-thickness perforation of the bronchial wall covered by a thick layer of a chronic inflammatory infiltrate. Infection by Candida at the bottom of some ulcerations; (2) Dumon stents: mild bronchial inflammation (squamous metaplasia, submucosal inflammatory infiltrates; granuloma-like infiltrates). In case of displacement, no significant changes of the previously stented bronchus occurred; and (3) Polyflex stents: no changes of the previously stented bronchi. CONCLUSIONS: Gianturco stents proved unsafe in the long term, owing to the risk of severe airway wall damage. The Polyflex stent is well tolerated but presents a high migration rate. Silicone stents show several limitations but appear to be well tolerated by the host mucosa. PMID- 10800790 TI - Differential effects of calcium channel antagonists in the amelioration of radial artery vasospasm. AB - BACKGROUND: Radial artery (RA) is being used for coronary artery bypass grafting (CABG) with greater frequency. However, RA is prone to post-CABG vasospasm, which may be neurohormonally mediated. Use of the calcium channel antagonist diltiazem has been advocated as a strategy to reduce post-CABG RA vasospasm. However, whether and to what degree different calcium channel antagonists influence neurohormonally induced RA vasoconstriction remains unknown. METHODS: RA segments were collected from patients undergoing elective CABG (n = 13), and isometric tension was examined in the presence of endothelin (10 nM) or norepinephrine (1 microM). In matched RA, endothelin- or norepinephrine-induced contractions were measured in the presence of diltiazem (277 nM), amlodipine (73 nM), or nifedipine (145 nM). These concentrations of calcium channel antagonists were based upon clinical plasma profiles. RESULTS: Endothelin and norepinephrine caused a significant increase in RA-developed tension (0.54+/-0.1 and 0.68+/-0.1 g/mg, respectively; p<0.05). Amlodipine or nifedipine significantly reduced RA vasoconstriction in the presence of endothelin (30+/-6% and 41+/-9%, respectively; p<0.05) or norepinephrine (27+/-8% and 53+/-9%, respectively; p<0.05), whereas diltiazem did not significantly reduce RA vasoconstriction. CONCLUSIONS: These results demonstrate that neurohormonal factors released post CABG can cause RA vasoconstriction, and that calcium channel antagonists are not equally effective in abrogating that response. Both amlodipine and nifedipine, which have a higher degree of vascular selectivity, appear to be the most effective in reducing RA vasoconstriction. PMID- 10800791 TI - Myocardial revascularization in the elderly using beating heart coronary artery bypass surgery. AB - BACKGROUND: Beating heart or "off-pump" coronary artery bypass (OP-CAB) has become an accepted method of myocardial revascularization by reducing the perioperative morbidity related to cardiopulmonary bypass (CPB). However, the efficacy of OP-CAB has not been well established in the elderly patient population. METHODS: OP-CABs were performed in 53 patients aged 75 years and older, at Pitt County Memorial Hospital from January 1996 to October 1999, either through a median sternotomy or an anterior thoracotomy. These results were compared with 220 patients who underwent standard coronary artery bypass graft (CABG) operation using CPB during the same time period. RESULTS: Mean patient age for both groups was 79+/-0.5 years and preoperative risk factors were similar. There were no differences in postoperative myocardial infarction, atrial fibrillation, bleeding, neurologic complications, or renal failure. There were no deaths after OP-CAB, compared with the 7.6% operative mortality rate after CABG (p<0.05). The OP-CAB group had a significantly shorter postoperative length of stay (4.4+/-0.4 days vs. 8.4+/-0.6 days) and lower transfusion requirements (0.4+/-0.1 units packed red blood cells vs 1.9+/-0.2 units packed red blood cells) than the CABG group. CONCLUSIONS: Our data demonstrate that OP-CAB is a safe and efficacious method of myocardial revascularization in the elderly, and may actually be preferential in these patients when applicable. PMID- 10800792 TI - Hypothermic circulatory arrest in octogenarians: risk of stroke and mortality. AB - BACKGROUND: The proportion of patients in their ninth decade of life undergoing complex cardiovascular procedures has increased over the past decade. The purpose of this study is to quantify the potential for stroke and mortality associated with deep hypothermic circulatory arrest (DHCA) in this age group. METHODS: At our institution, 251 adult patients had cardiovascular procedures that required DHCA since 1989. This included 20 patients 80 years of age or older (group I) and 231 patients less than 80 years (group II). Additionally, we analyzed 632 patients 80 years of age or older who underwent a variety of cardiovascular procedures since 1989 that required cardiopulmonary bypass but not DHCA (group III). Neurologic outcomes have been maintained in our database prospectively since 1991. RESULTS: The 30-day mortality in group I was 5%, in group II 15.2%, and in group III 8.2%. The stroke rate was 20% in group I, 8.8% in group II, and 6.5% in group III. CONCLUSIONS: DHCA can be performed with acceptable early mortality in patients in their ninth decade of life, but they are at an increased risk of stroke. Follow-up shows satisfactory late survival. PMID- 10800793 TI - Stroke after coronary artery operation: incidence, correlates, outcome, and cost. AB - BACKGROUND: Stroke is a major complication of coronary operation, with reported rates of postoperative cerebral dysfunction ranging from 0.4% to 13.8%. In this report, the incidence, correlates, outcomes, and costs of stroke in coronary operation were evaluated at Emory University between 1988 and 1996. METHODS: Data were entered prospectively into a dedicated computerized database at Emory University and analyzed retrospectively. Univariate and multivariate analyses were utilized where appropriate. RESULTS: Data from 10,860 patients undergoing primary coronary operation between 1988 and 1996 were analyzed. There were 250 patients not entered into the database. Stroke occurred in 244 (2.2%). Univariate predictors of stroke (p<0.05) included age, female gender, hypertension, diabetes, prior stroke, prior transient ischemic attack (TIA), and carotid bruits. Multivariate correlates included age (odds ratio 1.07) previous TIA (odds ratio 2.2), and carotid bruits (odds ratio 1.9), although the area under the Receiver Operating Characteristics (ROC) curve was only 0.69, suggesting limited ability to predict stroke. One and 5 year survival rates were 64% and 44% with stroke, and 94% and 81% without stroke, respectively. Among the stroke group, 23% of the patient population died before hospital discharge. The stroke group had a significantly longer length of hospital stay, as well as higher costs. CONCLUSIONS: Stroke is a devastating complication of coronary operation, significantly increasing morbidity, mortality, and cost. Three independent variables were identified for predicting stroke, including age, previous TIA, and carotid bruits. Patients should be carefully screened for cerebrovascular disease to help prevent stroke and its associated morbidity. PMID- 10800794 TI - Ineffectiveness and potential proarrhythmia of atrial pacing for atrial fibrillation prevention after coronary artery bypass grafting. AB - BACKGROUND: Atrial pacing is often used empirically to suppress atrial ectopy and prevent atrial fibrillation after coronary artery bypass grafting. METHODS: To determine whether atrial overdrive pacing reduces atrial fibrillation and atrial ectopy after coronary artery bypass grafting, 100 patients were randomized to no atrial pacing (Control) versus AAI pacing at 10 beats/min or more above the resting heart rate (Paced), started by postoperative day 1 and continued through day 4. Major end points were new atrial fibrillation and frequency of atrial ectopy during the first 4 days after coronary artery bypass grafting. RESULTS: Atrial fibrillation occurred by day 4 in 13 of 51 (25.5%) Paced and in 14 of 49 (28.6%) Control patients, p = 0.90. Control patients who developed atrial fibrillation had significantly more atrial ectopy than those who did not. Atrial ectopy was paradoxically more frequent in the Paced group (2,106+/-428 versus 866+/-385 per 24 hours, p = 0.0001). Loss of capture, sensing, and consistent atrial pacing occurred frequently during atrial pacing. CONCLUSIONS: Contrary to prevailing opinion and practice, postoperative atrial overdrive pacing significantly increases atrial ectopy and does not reduce the likelihood of atrial fibrillation. PMID- 10800795 TI - Tachyarrhythmias and triggering factors for atrial fibrillation after coronary artery bypass operations. AB - BACKGROUND: We evaluated the role of supraventricular arrhythmias and assessed clinical predictors of atrial fibrillation (AF) that developed after coronary artery bypass operations. METHODS: Eighty patients, with a mean age of 65.8 years, underwent 24-hour Holter monitoring preoperatively and for 4 consecutive days postoperatively, or until clinically documented AF, for analysis of the number of premature beats and tachyarrhythmias. Atrial areas and atrial peptides were measured preoperatively and postoperatively. RESULTS: Twenty-nine of 80 (36.3%) patients had postoperative AF. Preoperatively, the maximal supraventricular premature beats per minute were higher in the AF group (p = 0.02). The body mass index and total amount of cardioplegia were lower (p = 0.02 and p = 0.006, respectively), and withdrawal of beta-blockers postoperatively more frequent (p = 0.001) in the AF group, but atrial areas and atrial peptides did not differ. CONCLUSIONS: Frequent supraventricular premature beats preoperatively may indicate a propensity for AF. A larger amount of cardioplegia during the cross-clamp period may reduce the risk of postoperative AF. Further studies are mandatory to clarify why patients with lower body mass index were more prone to AF. PMID- 10800796 TI - Intraoperative physiologic variables and outcome in cardiac surgery: Part I. In hospital mortality. AB - BACKGROUND: Risk stratification schemes have been developed to predict outcome of coronary artery bypass grafting (CABG) procedures, which are predominately based upon unalterable preoperative patient characteristics. The purpose of this study was to determine if minimum intraoperative hematocrit, maximum glucose concentration, mean arterial pressure on cardiopulmonary bypass, or duration of bypass influence risk-adjusted in-hospital mortality after CABG. METHODS: Outcome data from 2,862 CABG patients were merged with intraoperative physiologic data. A preoperative mortality risk index was calculated for each patient. Variables found significant (p<0.05) by univariate logistic regression were tested in a multiple variable model to determine risk-adjusted association with mortality. RESULTS: Overall mortality rate was 1.85%. The preoperative risk index was significantly associated with mortality (p = 0.0001). No significant association was present between mortality and intraoperative variables. Preexisting hypertension was an independent predictor of mortality after controlling for risk index and bypass duration. CONCLUSIONS: Preexisting hypertension proved to be an independent predictor of mortality in our patient population. This study found no evidence to support the hypothesis that mean arterial pressure less than 50 mm Hg, lower hematocrit, or elevated glucose while on bypass increases in-hospital mortality. PMID- 10800797 TI - Intraoperative physiologic variables and outcome in cardiac surgery: Part II. Neurologic outcome. AB - BACKGROUND: The impact of alterable physiologic variables on neurologic outcome after coronary artery bypass grafting procedures is unknown. The purpose of this study was to determine whether minimum intraoperative hematocrit, maximum glucose concentration, or mean arterial pressure during cardiopulmonary bypass influences risk-adjusted neurologic outcome after coronary artery bypass grafting. METHODS: Outcome data from 2,862 patients undergoing coronary artery bypass grafting were merged with intraoperative physiologic data. A preoperative stroke risk index was calculated for each patient. Variables found significant by univariate logistic regression were tested in a multivariable model to determine association with outcome. RESULTS: The incidence of stroke or coma in the study population was 1.3%. After controlling for stroke risk and bypass time, only an index of low mean arterial pressure during bypass retained a significant inverse association with outcome (p = 0.0304). CONCLUSIONS: This study found no evidence that glucose concentration or minimum hematocrit are associated with major adverse neurologic outcome. The association between lower pressure during bypass and decreased incidence of stroke or coma persisted in all risk groups. This points to mechanisms other than hypoperfusion as the primary cause of neurologic injury associated with cardiac surgery. PMID- 10800798 TI - Cytokine release and neutrophil activation are not prevented by heparin-coated circuits and aprotinin administration. AB - BACKGROUND: Cardiopulmonary bypass (CPB) initiates a whole-body inflammatory response where complement and neutrophil activation and cytokine release play an important role. This prospective trial examined the effects of both heparin coated circuits and aprotinin on the inflammatory processes during CPB, with respect to cytokine release and neutrophil activation. METHODS: Two hundred patients undergoing cardiac surgery were randomized in four groups of 50 patients each: heparin-coated circuit with aprotinin (HCO-A) or without aprotinin (HCO) administration, and uncoated circuit with aprotinin (C-A) or without aprotinin administration (C). In groups receiving aprotinin, a high-dose regimen was given. In all groups, high initial doses of heparin were used (3 mg/kg intravenously). Tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and IL-8, and myeloperoxidase and elastase levels were measured in plasma samples taken before, during, and after CPB. RESULTS: In all groups, the TNF-alpha, IL-6, and IL-8 levels reached a maximum after protamine administration. After 24 hours, they remained significantly elevated (IL-6 and IL-8) or returned to baseline values (TNF-alpha). A similar pattern was observed with myeloperoxidase and elastase levels. No significant intergroup differences were observed. CONCLUSIONS: CPB is associated with cytokine release and neutrophil activation, which are not attenuated by the use of heparin-coated circuits or by the administration of aprotinin. Aprotinin and heparin-coated circuits do not show additive effects. PMID- 10800799 TI - Resource utilization in coronary artery bypass operation: does surgical risk predict cost? AB - BACKGROUND: Current healthcare trends may render financial risk of cardiac operation a key component of clinical decision making. It has been suggested, based on large cohorts of patients stratified by clinical risk, that the cost of operation can be predicted from models of clinical risk since length of stay (LOS) is highly correlated to clinical risk, and LOS is correlated to hospital costs and charges. Direct correlation of actual surgical costs with surgical risk are lacking. METHODS: Variable direct costs, LOS, and The Society of Thoracic Surgeons predicted mortality risk [STS risk (%)] were collected and analyzed in 628 consecutive patients undergoing coronary artery bypass grafting (CABG) at our institution in 1997. RESULTS: Cost of CABG had a near-normal distribution, and cost in 21 outlier patients (cost > two standard deviations above the mean) was an average 5.3 times normal (median cost). For individual patients, cost was well correlated to LOS (R2 = 0.48) but not with STS risk (R2 = 0.12). LOS was also poorly predicted by STS risk (R2 = 0.09). However, despite its poor prediction of cost, STS risk was an unbiased estimator over the entire population. A result manifested, when patients were grouped into similar risk (<1%, 1-2%, 2+ -3%, 3+ 5%, 5+ -10%, and >10%) cohorts, by high correlation between cost and STS risk (R2 = 0.99), cost and LOS risk (R2 = 0.99), and LOS and STS risk (R2 = 0.97). CONCLUSIONS: Our data demonstrated that, in large CABG cohorts, surgical risk models can accurately predict cost of CABG. However, despite a trend for increasing cost with increasing STS risk, surgical risk models based on preoperative data are poor predictors of cost in individual patients. Use of these models should be limited to analysis of cost trends in cardiac operation, but not for predicting financial risk in individual patients during clinical decision making. PMID- 10800800 TI - Perioperative cardiac function and predictors for adverse events after transmyocardial laser treatment. AB - BACKGROUND: Previous studies have reported that mortality and morbidity after transmyocardial laser treatment (TML) mainly occur perioperatively. The present study was designed to evaluate left-ventricular function and identify risk factors for cardiac-related adverse events in this phase. METHODS: Forty-nine patients were studied. The inclusion criteria were angina pectoris Canadian Cardiovascular Society Angina Score (CCSAS) class III and IV refractory to medical therapy and untreatable by coronary artery bypass graft or percutaneous transluminal coronary angioplasty, age less than 75 years, left ventricular ejection fraction greater than or equal to 30%, and myocardial regions with reversible ischemia. Hemodynamic data and cardiac adverse events were registered. The follow-up time was 30 days. RESULTS: A transient decrease in mean cardiac index (CI) was observed, reaching its minimum immediately after end of the surgical procedure (1.8+/-0.4, p<0.01 vs. baseline). Two patients (4%) died during the postoperative period (30 days). Seventeen patients (35%) experienced adverse cardiac-related events, where CCSAS class IV, unprotected left main stem stenosis, and diabetes mellitus were identified as risk factors in a multivariate analysis. CONCLUSIONS: A transient impairment of left ventricular function was observed after TML. The morbidity and mortality after TML were almost exclusively cardiac-related, identifying CCSAS class IV, unprotected left main stem stenosis, and diabetes as risk factors. PMID- 10800801 TI - Coagulase-negative staphylococci and sternal infections after cardiac operation. AB - BACKGROUND: Coagulase negative staphylococci (CoNS) have been recognized as important pathogens in nosocomial infections, especially in connection with implanted foreign materials. In cardiac operation they are among the most common pathogens isolated from infected sternal wounds. The definition of the infection is very important. In this study we focus on deep postoperative chest infections. METHODS: By studying 33 infected patients retrospectively and comparing them to 33 matched uninfected controls, we studied the characteristics and costs of the infections. RESULTS: Typical for these infections is the late and insidious onset, and that the infections initially give only minor symptoms such as pain, redness, and serous secretion. We found the following risk factors for infection: number of preoperative days in a hospital, the total length of the operation, and if the patient had undergone an early reoperation due to causes other than infection. This kind of infection more than doubled the hospital costs for the patients affected. CONCLUSIONS: Coagulase negative staphylococci are the most important pathogens in deep postoperative infections in this material. They cause infections that are difficult to recognize since they give only discrete symptoms and start well after the patients leave the hospital. The risk factors for patients with CoNS infections are mostly associated with a long exposure to the hospital environment. The treatment is often difficult and costly because of multiresistant bacteria and frequent need for repeated surgical revisions. PMID- 10800802 TI - Spread of coagulase-negative staphylococci during cardiac operations in a modern operating room. AB - BACKGROUND: Coagulase-negative staphylococci cause 33% to 62.5% of wound infections after cardiac operations. The aim of this study was to investigate the sources of coagulase-negative staphylococci in the sternal wound. METHODS: Twenty operations performed in zonal ventilated operating rooms were investigated prospectively. Cultures were taken from all persons present in the room, the sternal wound, and the air. Isolates macroscopically judged to be coagulase negative staphylococci were metabolically classified, and similar isolates were investigated by pulsed-field gel electrophoresis. RESULTS: Bacterial counts in the operating room air were very low. Wound contamination was found in 13 of 20 operations. Six wound isolates could be traced, three to the patients' sternal skin, one to the patient's groin, one to the surgeon's nose, and one to the surgeon's arm and forehead and the assistant's nose. Three operating field air cultures could be traced to the scrubbed theatre staff. The single case of superficial sternal wound infection was caused by Staphylococcus aureus, which was not isolated from the wound at operation. CONCLUSIONS: In an ultraclean environment, bacteria in the sternal wound originated from the patients' own skin and from the surgical team. PMID- 10800803 TI - Functional properties of the saphenous vein harvested by minimally invasive techniques. AB - BACKGROUND: Since surgical techniques affect the functional properties of the vessel wall, the present study investigated the influence of minimally invasive harvesting techniques on the vascular reactivity of the saphenous vein. METHODS: Saphenous vein remnants were obtained after aortocoronary bypass operation from patients subjected to conventional (n = 6), mediastinoscope-assisted (n = 4), or endoscope-assisted venectomy (n = 5). After preservation in University of Wisconsin solution (UW), ring preparations were mounted in a standard organ bath setup and concentration-response curves were constructed for phenylephrine, sodium nitroprusside, and acetylcholine. RESULTS: Saphenous vein reactivity was not altered after preservation in UW. For the vein preparations harvested by means of the three venectomy methods, no differences were demonstrated for responses to KCl, phenylephrine, or sodium nitroprusside. The maximal endothelium dependent acetylcholine-induced dilation of precontracted vein rings varied between 5% and 12%, independent of the surgical technique applied. CONCLUSIONS: It was demonstrated that minimally invasive surgical techniques for harvesting the saphenous vein, which are developed to reduce postoperative complications at the site of explantation, did not affect the vascular reactivity in a different manner than the conventional method. PMID- 10800804 TI - Ulinastatin attenuates reperfusion injury in the isolated blood-perfused rabbit heart. AB - BACKGROUND: Ventricular dysfunction after long cardioplegic arrest has been observed in cardiac operations. Urinary trypsin inhibitor, also called ulinastatin, may attenuate myocardial ischemia-reperfusion injury. The present study was designed to determine the protective efficacy of ulinastatin in blood perfused parabiotic isolated rabbit hearts as a surgically relevant model with long (4-hour) cardioplegic arrest. METHODS: Each isolated rabbit heart, with a latex balloon inserted in the left ventricle, was parabiotically blood-perfused using a modified Langendorff column. The left ventricular developed pressure, rate of pressure development, and coronary flow with a left ventricular end diastolic pressure of 10 mm Hg were measured before ischemia and 15, 30, 45, and 60 minutes after reperfusion began (control, n = 10). Ulinastatin (15,000 U/kg) was administered to the support animal just before reperfusion began (group U-1, n = 10) or at the beginning of the extracorporeal circulation and readministered before reperfusion (group U-2, n = 10). The endothelium of the coronary artery was observed by scanning electron microscopy to evaluate the extent of endothelial ischemia-reperfusion injury. RESULTS: Ulinastatin enhanced the recovery of developed pressure in both the U-1 (p<0.05) and U-2 (p < 0.01) groups compared with the control group. Although ulinastatin given just before reperfusion (group U-1) did not enhance the recovery of the rate of pressure development or the coronary flow compared with the control, earlier administration did improve the recovery of the rate of pressure development compared with the control (U-2, p<0.05), and there was improvement of the recovery of coronary flow after 60 minutes of reperfusion (U-2, p<0.05). Scanning electron microscopy showed that ulinastatin had ameliorated coronary endothelial damage. CONCLUSIONS: Ulinastatin improved functional recovery after long cardioplegic arrest and reduced coronary endothelial injury. Administration of ulinastatin at the beginning of cardiopulmonary bypass and just before reperfusion may be useful clinically in cases requiring prolonged aortic cross clamping. PMID- 10800805 TI - Axillary artery cannulation in acute ascending aortic dissections. AB - BACKGROUND: Standard cannulation of the femoral artery in preparation for repair of a dissection involving the ascending aorta carries a high risk of malperfusion. Arterial perfusion through the right axillary artery is more likely to perfuse the true lumen and should be advantageous in acute dissections involving the ascending aorta. METHODS: Thirteen patients underwent repair of acute ascending aortic dissections and were perfused through the right axillary artery. All had deep hypothermic circulatory arrest. RESULTS: There was one mild intraoperative cerebrovascular accident with complete recovery and one operative death secondary to low cardiac output. There were no intraoperative problems with perfusion through the axillary artery, and there were no postoperative problems or complications involving the axillary artery, axillary vein, or brachial plexus. CONCLUSIONS: Arterial perfusion through the right axillary artery is a safe and effective means of more reliably perfusing the true lumen. In this regard, it may be superior to femoral artery perfusion and could lead to improved outcomes with repair of acute deBakey type I and II aortic dissections. PMID- 10800806 TI - Effect of cerebral embolization on regional autoregulation during cardiopulmonary bypass in dogs. AB - BACKGROUND: Embolization during cardiopulmonary bypass probably alters cerebral autoregulation. Therefore, using laser Doppler flowmetry we investigated the cerebral blood flow velocity changes in response to changes in arterial pressure, before and after embolization in a canine bypass model. METHODS: After Institutional Animal Care and Use Committee approval, 8 anesthetized dogs had a laser Doppler flow probe positioned over the temporoparietal dura. During 37 degrees C cardiopulmonary bypass, the cerebral blood flow velocity response to changing mean arterial pressure (40 to 85 mm Hg in random order) was assessed before and after systemic embolization of 100 mg of 97-microm latex microspheres. RESULTS: Before embolization, cerebral blood flow velocity increased 39% as mean arterial pressure increased from 40 to 85 mm Hg. Following embolization, a 94% increase in cerebral blood flow velocity was demonstrated over the same mean arterial pressure range. The slopes of the curves relating cerebral blood flow velocity to mean arterial pressure were 0.21+/-0.74 and 1.31+/-0.87, before and after embolization (p = 0.016) respectively. CONCLUSIONS: Regional cerebral blood flow autoregulation may be impaired by microembolization known to occur during cardiopulmonary bypass, increasing the dependence of cerebral blood flow on mean arterial pressure. PMID- 10800807 TI - Coronary artery bypass grafting in patients who require long-term dialysis. AB - BACKGROUND: Should coronary artery bypass grafting (CABG) be performed in patients on long-term dialysis? This subject has been debated for several years. We retrospectively reviewed the charts of all patients who had CABG from August 1989 to October 1997. METHODS: We identified 70 patients who were on long-term dialysis and had CABG during that time period. Patients were evaluated by chart review and telephone survey. Forty-nine patients (70%) had unstable angina and 37 patients (52%) had triple vessel disease. Patient risk factors included 60 patients with hypertension (85%), 40 patients with diabetes mellitus (57%), 35 patients who had congestive heart failure (50%), 35 patients who had a previous myocardial infarction (50%), and 31 smokers (44%). Operative procedures included 49 patients who had CABG only and 21 patients who had concomitant CABG with valve replacement or repair. During the postoperative period, complications developed in 50% of patients. RESULTS: Review of these complications showed that 25% of patients required prolonged mechanical ventilation, and 10% of patients had septicemia. Operative mortality was high, with 10 patient deaths (14.3%) within 30 days of the procedure. Six (60%) of these deaths occurred in patients who had CABG and valve repair or replacement. Long-term follow up at 50.3 months showed no improvement in survival in patients who had CABG compared with the known mortality rate of 22% per year in dialysis patients regardless of comorbid conditions. Quality of life subjectively improved in only 41% of patients in follow-up telephone survey. CONCLUSIONS: Patients requiring long-term dialysis with coexistent severe cardiac disease should be thoroughly evaluated preoperatively. One must weigh the high morbidity and mortality risk against the limited long-term resolution of angina and ultimate survival. PMID- 10800809 TI - Left ventricular assist device bridge therapy for acute myocardial infarction. AB - BACKGROUND: Patients with acute myocardial infarction (AMI) complicated by cardiogenic shock have a high mortality rate. Current treatment modalities remain suboptimal for these patients. METHODS: From April 1995 to March 1998, 7 patients were identified as having AMI associated with cardiogenic shock. All received intraaortic balloon pump assistance, in addition to maximal inotropic support. RESULTS: The mean preoperative cardiac index was 2.0+/-0.3 L/min/m2 and pulmonary capillary wedge pressure was 23+/-6 mm Hg. Three patients received thrombolytic therapy and 4 patients underwent percutaneous transluminal coronary angioplasty without success. Left ventricular assist devices (LVADs) were implanted as bridge therapy to heart transplantation. One patient died from recurrence of a ventricular septal defect during LVAD support. Six patients were transplanted successfully after mean LVAD support of 59+/-33 days. Five patients are alive and well at a mean follow-up of 898+/-447 days. One patient died 3 days after transplantation from acute allograft dysfunction. CONCLUSIONS: Timely application of LVADs as bridge therapy to heart transplantation in these critically ill patients can be lifesaving, and should be investigated further. PMID- 10800808 TI - Beating heart surgery in octogenarians: perioperative outcome and comparison with younger age groups. AB - BACKGROUND: Octogenarians have higher morbidity and mortality rates (9% to 16%) after coronary artery bypass grafting with cardiopulmonary bypass, compared with younger patients. METHODS: We compared the perioperative outcome and hospital stay after coronary artery bypass grafting without cardiopulmonary bypass (off pump) from January 1987 to May 1999, among patients older than 80 years (n = 71), patients between 70 and 79 years (n = 228), and patients whose age ranged from 60 to 69 years (n = 296). In comparison with younger patients, more octogenarians were female (51% versus 39% in patients aged 70 to 79 years and 35% in those aged 60 to 69 years, p = 0.04), they had previous myocardial infarction more frequently (48% versus 47% versus 34%, respectively, p = 0.008), and were operated on urgently (69% versus 56% versus 52%, respectively, p = 0.04). RESULTS: Postoperative complications that were significantly higher in octogenarians compared with younger groups included pneumonia (6% in octogenarians versus 2% in patients aged 70 to 79 years and 0% in patients aged 60 to 69 years, p = 0.001) and atrial fibrillation (47% versus 32% versus 21%, respectively, p<0.001). By multivariate logistic regression analysis, age over 80 years was an independent predictor of prolonged hospital stay (odds ratio = 2.7, 95% confidence interval, 1.4 to 5, p<0.001). The in-hospital mortality rate was higher in octogenarians (6% versus 3% for 70 to 79 year-olds and 0.3% for 60 to 69 year-olds, p = 0.006). CONCLUSIONS: When appropriately applied in patients older than 80 years, off-pump coronary artery bypass grafting can be done with acceptable postoperative morbidity, mortality, and hospital stay. PMID- 10800810 TI - Is early tracheostomy safe in cardiac patients with median sternotomy incisions? AB - BACKGROUND: Tracheostomy after cardiac operation through a median sternotomy is believed to increase the risk of mediastinitis, leading to debate as to whether early tracheostomy is safe in these patients. METHODS: A record search of patients undergoing cardiac operation through median sternotomy was done. Day and duration of tracheostomy were correlated to day of positive bacteriological evidence and clinical outcome for the patient. The method of tracheostomy was also recorded. RESULTS: Of 174 cases, 4 patients had mediastinitis, 3 before tracheostomy was performed. Of these three patients, 2 survived and the third died of multiorgan failure 46 days after the procedure. The fourth patient, on immunosuppressive therapy for severe rheumatoid arthritis and pulmonary fibrosis, had tracheostomy performed at primary operation, developed fatal mediastinitis after 6 days, and died 18 days postoperatively of multiorgan failure. Of the tracheostomies performed, 24 (14%) were percutaneous, and 110 (63%) were achieved using standard surgical techniques (in 40 cases type was unrecorded). In 72 cases (41%), tracheostomy was performed on or before day 7, 11 (6%) being performed before 48 hours. Mortality occurred in 38 (22%). CONCLUSIONS: There is no demonstrable relationship between early tracheostomy and mediastinitis in median sternotomy patients. PMID- 10800811 TI - Therapeutic angiogenesis with intramyocardial administration of basic fibroblast growth factor. AB - BACKGROUND: Basic fibroblast growth factor (bFGF) induces endothelial cell and smooth muscle cell proliferation and stimulates angiogenesis. This study was designed to evaluate the effects of intramyocardial administration of bFGF on myocardial blood flow, angiogenesis, and ventricular function in a canine acute infarction model. METHODS: Myocardial infarction was induced in 12 dogs by ligation of the left anterior descending coronary artery. Within 5 minutes after coronary occlusion, 100 microg of human recombinant bFGF in 1 mL of saline was injected into the infarct and border zone in 6 dogs, whereas saline alone was used in 6 control dogs. Myocardial blood flow was determined with colored microspheres before and immediately after coronary ligation and again 3, 7, 14, and 28 days after treatment and it was expressed as percent of normal. Angiogenesis was evaluated by immunohistochemical studies 28 days later. Cardiac function was evaluated by repeated echocardiographic measurement. RESULTS: Treatment with bFGF significantly increased the endocardial blood flow in the border zone (7 days after infarction, 75%+/-7% and 41% +/-7% in the bFGF and control groups, respectively, p<0.01) as well as epicardial blood flow in the infarcted zone. Treatment with bFGF significantly increased the capillary density (39.7+/-2.3 and 22.7+/-1.1 vessels per visual field in the bFGF and control groups, respectively, p<0.01) as well as arteriolar density in the border zone. Treatment with bFGF significantly reduced the change in ratio of thickness of the infarcted wall to the normal wall (44%+/-6% and 26% +/-5% in the bFGF and control groups, respectively, p<0.05). It improved the left ventricular ejection fraction (7 days after infarction, 0.54+/-0.02 and 0.37+/-0.03 in the bFGF and control groups, respectively, p<0.01). CONCLUSIONS: Intramyocardial administration of bFGF increased the regional myocardial blood flow, reduced thinning of the infarcted region, and improved ventricular function in acute myocardial infarction. Intramyocardial administration of bFGF may be a new therapeutic approach for patients with acute myocardial infarction. PMID- 10800812 TI - Neuromonitoring and neurocognitive outcome in off-pump versus conventional coronary bypass operation. AB - BACKGROUND: Cardiopulmonary bypass seems to be a major cause for both intraoperative microemboli and cerebral hypoperfusion. This study investigates high intensive transient signals (HITS) in transcranial Doppler ultrasound (TCD) and serum levels of the neurobiochemical marker protein S-100 in patients who underwent coronary artery bypass operation without cardiopulmonary bypass (off pump CABG) in comparison with the conventional procedure using cardiopulmonary bypass (CPB). The results are related to the neuropsychologic outcome in both surgical groups. METHODS: Forty patients were randomized in 2 groups (20 conventional and 20 off-pump CABG). Neurocognitive status was assessed preoperatively and postoperatively. Venous serum levels of S-100 protein were measured before and after coronary operation, HITS were measured in the middle cerebral artery during the operation. RESULTS: The median value of HITS was 394.5 (0 to 2217) in the conventional versus 11 (0 to 50) in the off-pump group, p less than 0.0001. Postoperative S-100 serum levels were: 3.76 (0.13 to 11.2) microg/L (conventional) versus 0.13 (0.04 to 1.01) microg/L (off-pump), p less than 0.0001. Postoperative cognitive testing showed significantly different results with a postoperative impairment of 90% of the patients in the conventional group versus no impairment in the off-pump group. CONCLUSIONS: Cognitive impairment seems to be strongly associated to CPB and the occurrence of micro-emboli. The off-pump technique appears to be promising in order to eliminate the source of these neuropyschologic impairments following CABG operation. PMID- 10800813 TI - 25-year experience with 456 combined mitral and aortic valve replacement for rheumatic heart disease. AB - BACKGROUND: Valvular heart disease in developing countries resulting from rheumatic fever is disabling and if untreated leads to congestive heart failure and death. Valve replacement has remained the procedure of choice for advanced valve disease. METHODS: Between 1973 and 1997, 456 patients underwent combined mitral and aortic valve replacement. In light of our favorable earlier experience the Starr-Edwards ball valve prostheses were implanted in 90% and 72.8% of mitral and aortic positions, respectively. Follow-up ranged from 1 to 24 years with a median of 8.5 years. RESULTS: The 30-day hospital death rate was 9.2% and late death occurred in 10.1%. A low-intensity anticoagulant regimen was followed to maintain the target prothrombin time at 1.5 times the control value. The actuarial survival at 5, 10, 20, and 24 years was 90.4%, 85.6%, 84.4%, and 82.4% per year, respectively. CONCLUSIONS: In view of the acknowledged advantage of superior durability, increased thromboresistance in our patient population, and its cost effectiveness the Starr-Edwards ball valve is the mechanical prosthesis of choice for advanced combined valvular disease. The low-intensity anticoagulant regimen has offered sufficient protection against thromboembolism as well as hemorrhage. PMID- 10800814 TI - Bleeding from the sternal marrow can be stopped using vivostat patient-derived fibrin sealant. AB - BACKGROUND: Median sternotomy is the most important method of access to the heart. Bleeding from the sternal marrow may become significant, especially in elderly patients. Vivostat (ConvaTec, a Bristol-Myers Squibb Company, Skillman, NJ) patient-derived fibrin sealant is biocompatible and easily applied to the sternal marrow using the Vivostat Spraypen applicator. METHODS: Thirty patients undergoing elective cardiac operation were randomized to receive Vivostat fibrin sealant applied to either the right or left side of the sternum immediately after median sternotomy, with the untreated side serving as control. RESULTS: The average time to hemostasis was 43 seconds after treatment with Vivostat and 180 seconds on the control sides (p<0.001). At the end of the operation, complete hemostasis was observed on 24 of 30 sides treated with Vivostat compared with on 4 of 30 of the control sides (p<0.001). The average volume of sealant used to cover one side of the sternum was 0.9 mL. CONCLUSIONS: Vivostat patient-derived fibrin sealant is a biocompatible alternative to bone wax, with the results of this study showing that it provides effective control of bleeding after median sternotomy. PMID- 10800815 TI - Three-dimensional video and robot-assisted port-access mitral valve operation. AB - BACKGROUND: In order to minimize surgical trauma, video-assisted mitral valve operation has been started using the Port-Access technique with the addition of a three-dimensional visualization system (Vista Cardiothoracic Systems Inc, Westborough, MA) and a voice-controlled camera-holding robotic arm (Aesop; Computer Motion Inc, Goleta, CA). METHODS: Port-Access mitral valve replacement or repair (PAMVR) was undertaken using an endovascular cardiopulmonary bypass (CPB) system. Fifty patients underwent Port-Access mitral valve replacement or repair. A three-dimensional thoracoscope was inserted allowing complete three dimensional projection of the mitral valve (Vista). In the last 20 patients, the camera was attached to a robotic arm (Aesop), which allowed stabilization and voice-activated movement of the camera. Mitral valve repair was performed in 26 patients, and the valve was replaced in 24 patients with a mechanical valve prosthesis. RESULTS: Median time of operation was 4.2 hours, aortic cross-clamp time 83 minutes, CPB time 125 minutes, intensive care unit stay 1.5 days and hospitalization 9.0 days. Three months follow-up was complete in 40 patients, with 34 patients (85%) in New York Heart Association class I and 6 patients in class II. Mortality was 0% and rate of reoperation was 2%, with a follow-up time up to 1.5 years postoperatively. CONCLUSIONS: Using three-dimensional video and robotic assistance, it was possible to minimize the length of skin incision, but at the same time to optimally visualize the whole mitral valve apparatus in order to perform true Port-Access mitral valve operation, including various repair techniques. PMID- 10800816 TI - Functional assessment of disease-free saphenous vein grafts at redo coronary artery bypass grafting. AB - BACKGROUND: Reoperations for coronary artery bypass grafting are on the rise. The general rule of replacing all saphenous vein grafts (SVGs) older than 5 years of age at the time of reoperation has recently been challenged on clinical grounds. This study provides functional data of endothelial behavior in long-term vein grafts. METHODS: Previously placed SVGs were removed at the time of redo operations. Nitric oxide (NO) measurements in real time were carried out before and after stimulation with morphine. The measurements were compared to the angiographic appearance of the grafts obtained prior to operation. Grafts were categorized into 3 groups: disease-free, moderately diseased, and severely diseased. RESULTS: Sixteen grafts were analyzed. Five were angiographically disease-free, 4 had moderate, and 7 severe disease. In the disease-free group, peak NO production after 10(-6) mol/L morphine stimulation was 35 mol/L, equivalent to the production of native saphenous vein. The severely diseased group did not demonstrate an increase in NO production, and the moderately diseased group produced a small rise in production. CONCLUSIONS: Measurement of NO release of old SVGs, when angiographically pristine, equals that of native saphenous vein. These findings support the recent clinical observations that long term angiographically disease-free vein grafts are biologically privileged. PMID- 10800817 TI - Hemodynamic changes and right heart support during vertical displacement of the beating heart. AB - BACKGROUND: Hemodynamic instability during heart displacement in off-pump multivessel coronary artery bypass grafting might be related to right heart dysfunction. The Enabler (HemoDynamics Systems Ltd, Upper Yoqneam, Israel) is a cannula pump that expels blood from the right atrium into the pulmonary artery. We studied the hemodynamic changes and the role of the enabler during heart displacement. METHODS: Nine anesthetized sheep were assessed for hemodynamic changes during 90-degree heart displacement with or without Enabler support. Hemodynamic parameters included cardiac output, systemic arterial blood pressures, and left and right heart filling pressures. RESULTS: Heart displacement caused a significant decrease in cardiac output and systemic blood pressure (46%+/-5%, p = 0.001; and 20%+/-5%, p = 0.009, respectively), with a concomitant 137%+/-24% (p = 0.003) increase in central venous pressure. No significant change in left atrial pressure was observed. Activation of the Enabler caused a significant increase in cardiac output and systemic blood pressure (67%+/-15%, p = 0.01; and 17%+/-7%, p = 0.04, respectively), as well as a decrease in central venous pressure by 49%+/-8% (p = 0.0001). CONCLUSIONS: Heart displacement causes hemodynamic instability mainly by right heart dysfunction. The Enabler significantly stabilized circulation during vertical displacement of the beating heart. PMID- 10800818 TI - Leukocyte integrin expression in patients undergoing cardiopulmonary bypass. AB - BACKGROUND: The recruitment of leukocytes to vascular endothelium is controlled by adhesion events mediated through the beta2 integrins, whereas the response of extravasated leukocytes within the tissues is controlled through the beta1 integrins. Although cardiopulmonary bypass (CPB) has been shown to be associated with a systemic inflammatory response and elevated levels of beta2 integrins on leukocytes, its effect on the beta1 integrins is not known. This study investigated the effect of the protease inhibitor aprotinin on the expression of the beta1 and beta2 integrins on circulating leukocytes in patients undergoing CPB. METHODS: Patients undergoing primary elective coronary artery bypass grafting were randomized into full-dose aprotinin or placebo groups. Blood samples were obtained at nine time points preoperatively, intraoperatively, and up to 6 days postoperatively. The surface expression of the beta1 integrins VLA 1, -3, -4, -5, and -6 and of the beta2 integrins CD11a/CD18, CD11b/CD18, and CD11c/CD18 was measured by flow cytometry on gated neutrophil and monocyte subpopulations in whole blood. RESULTS: Expression of the beta1 integrins was not significantly altered during the study period and, therefore, aprotinin had no effect on the expression of these molecules. Of the beta2 integrins, CD11b/CD18 expression was significantly increased on neutrophils at 15 minutes after onset of CPB in the placebo group (p < 0.01) but not in the aprotinin group. CONCLUSIONS: This study showed that expression of the beta1 integrins on neutrophils and monocytes did not alter during the first 6 days after CPB. Expression of the beta2 integrin CD11b/CD18 increased significantly on neutrophils during CPB in control patients but not in patients treated with full dose aprotinin. PMID- 10800820 TI - Clinical and pathological features of accessory valve tissue. AB - BACKGROUND: Accessory valve tissue is a rare congenital cardiac malformation causing a variety of clinical features. We describe our experience with the surgical treatment of accessory valve tissue to clarify clinical and pathological features of this anomaly. METHODS: Between December 1990 and February 1999, 11 children underwent resection of accessory valve tissue together with repair of coexisting intracardiac malformations. Eight patients had accessory tricuspid valve, while 3 had accessory mitral valve tissue. Associated intracardiac malformations included tetralogy of Fallot in 6 patients, transposition of the great arteries in 3, coarctation complex in 1, and ventricular septal defect in 1. RESULTS: Accessory valve tissue was completely resected in all but 1 patient with accessory mitral valve and transposition of the great arteries. As total resection of the accessory valve tissue through the pulmonary artery was unsuccessful, partial resection and bidirectional Glenn shunt operation was performed instead of arterial switch operation in this patient. CONCLUSIONS: The clinical signs and symptoms of accessory valve tissue vary according to its location and coexisting cardiac malformations. Congenital heart surgeons should be familiar with the clinical and pathological features of accessory valve tissue, so as not to overlook it during operation. PMID- 10800819 TI - Inflammatory response after coronary revascularization with or without cardiopulmonary bypass. AB - BACKGROUND: We sought to investigate the effect of multiple coronary artery bypass grafting (CABG) with or without cardiopulmonary bypass (CPB) on the perioperative inflammatory response. METHODS: Sixty patients undergoing CABG were randomly assigned to one of two groups: (A) on pump with conventional CPB and cardioplegic arrest, and (B) off pump on the beating heart. Serum samples were collected for estimation of neutrophil elastase, interleukin 8 (IL-8), C3a, and C5a preoperatively and at 1, 4, 12, and 24 hours postoperatively. Furthermore, white blood cell (WBC), neutrophil, and monocyte counts were carried out preoperatively and at 1, 12, 36 and 60 hours postoperatively. Overall incidence of infection and perioperative clinical outcome were also recorded. RESULTS: The groups were similar in terms of age, weight, gender ratio, extent of coronary disease, left ventricular function, and number of grafts per patient. Neutrophil elastase concentration peaked early after CPB in the on-pump group, with a decline with time. Repeated-measures analysis of variance between groups and comparisons at each time point (modified Bonferroni) showed elastase concentrations were significantly higher in the on-pump than the off-pump group (both p < 0.0001). IL-8 increased significantly after surgery in the on-pump group, with no decline during the observation period (p = 0.01 vs off pump). C3a and C5a rose early after surgery in both groups when compared with baseline values. Postoperative WBC, neutrophil, and monocyte counts were significantly higher in the on-pump than the off-pump group (p < 0.01). Finally, the incidence of postoperative overall infections was significantly higher in the on-pump group (p < 0.0001 vs off pump). CONCLUSIONS: CABG on the beating heart is associated with a significant reduction in inflammatory response and postoperative infection when compared with conventional revascularization with CPB and cardioplegic arrest. PMID- 10800821 TI - Isolated congenital absence of the pericardium: clinical presentation, diagnosis, and management. AB - BACKGROUND: Congenital absence of the pericardium (CAP) is a rare clinical entity. METHODS: We identified from the two hospital databases all patients with isolated CAP, reviewed their data, and invited them for prospective clinical evaluation with electrocardiography, chest x-ray findings (CXR), echocardiography, and magnetic resonance imaging (MRI). RESULTS: Ten patients (3 males, 7 females) presented at a median age of 21 years (range, 2-53 years) with paroxysmal stabbing chest pain, largely nonexertional (9), and heart murmur with an abnormal CXR (1). Three patients had partial and 7 had complete CAP (all 7 had marked lateral displacement of the cardiac apex). CXR combined with MRI were key to establishing the diagnosis; a "tongue" of lung tissue interposing between the main pulmonary artery and aorta was the most consistent diagnostic feature. Four patients underwent pericardioplasty, 3 for debilitating symptoms and 1 for left atrial appendage herniation, followed by improvement or resolution of symptoms. At a mean of 10.5 years from presentation all patients were alive. No complications were seen in the nonsurgical group. CONCLUSIONS: Isolated CAP has a common presentation pattern with periodic stabbing chest pain mimicking coronary artery disease. CXR and MRI are required for definitive diagnosis. Symptomatic patients with the complete form may benefit from pericardioplasty. PMID- 10800822 TI - Surgical myocardial revascularization without cardiopulmonary bypass. AB - BACKGROUND: Though coronary artery bypass grafting (CABG) without cardiopulmonary bypass is being performed with increasing frequency, in the absence of adequate angiographic follow-up, safety, reproducibility, and efficacy of the procedure remain doubtful. In this prospective study, we report the results obtained by 100% angiographic follow-up of 96 consecutive patients. METHODS: A total of 96 patients (age range 33 to 76 years) underwent CABG without cardiopulmonary bypass. Single vessel disease was present in 46 (47.9%) patients, double vessel disease in 31 (32.3%), and triple vessel disease in 19 (19.8%) patients. All patients were operated through a standard midsternotomy and an optimal combination of pharmacological and mechanical methods were used to restrict cardiac movements during anastomosis. All patients underwent coronary angiography before discharge from the hospital. RESULTS: A total of 160 grafts were placed (range 1 to 4 grafts per patient, average 1.7+/-0.3 grafts per patient). A single graft was placed in 46 patients, double grafts in 38, triple grafts in 10, and quadruple grafts in 2 patients. Various grafts included pedicled left internal mammary artery (LIMA) (n = 95), free LIMA (n = 1), right internal mammary artery (n = 14), radial artery (n = 24), right gastroepiploic artery (n = 5), and saphenous vein grafts (n = 21). Operative mortality was 1.0% (1 of 96). Two patients required reoperation for excessive bleeding. Mean hospital stay was 5.7+/-1.2 days. Overall angiographic patency was 95.0% with LIMA patency of 97.9% (93 of 95). One patient with block in midsegment of LIMA was reoperated using cardiopulmonary bypass. Follow-up ranged from 4 to 17 months (mean 8.2+/-3.1 months). Two patients (one with narrowed LIMA to left anterior descending artery anastomosis, and one with patent anastomosis) had residual angina. CONCLUSIONS: Coronary artery bypass grafting without cardiopulmonary bypass is a reproducible, effective, and safe option in selected group of patients. A conscientious approach in patient selection and route of operation is required. PMID- 10800823 TI - Incidence and implications of systemic to pulmonary collaterals after bidirectional cavopulmonary anastomosis. AB - BACKGROUND: Systemic to pulmonary arterial collaterals often develop after bidirectional cavopulmonary anastomosis (BCPA). It has been proposed that such collaterals may be related to perioperative outcome and duration of effusions after the modified Fontan procedure. However, the incidence and significance of collaterals after BCPA remain uncertain. METHODS: To evaluate risk factors for, and significance of, such collaterals, we reviewed angiographic and clinical data for all 76 patients who underwent BCPA between January 1990 and June 1996 and had follow-up catheterization during or before 1997. RESULTS: The median age at BCPA was 10 months, and the median duration from BCPA to follow-up catheterization was 18 months. Arterial collaterals were detected on follow-up catheterization in 45 patients (59%). Factors associated with collateral development included a prior right-sided systemic-to-pulmonary arterial shunt, lower pre-BCPA end-diastolic ventricular pressure and pulmonary vascular resistance, and use and duration of cardiopulmonary bypass during the BCPA operation. Fourteen of the 45 patients (30%) underwent coil embolization of the collaterals. Forty-three patients have undergone extracardiac conduit Fontan, with 1 early and 1 late death. Collaterals were present in 22 of these patients, 7 of whom underwent pre-Fontan embolization. The duration from BCPA to Fontan was longer in patients with collaterals, but these patients were not more likely to have prolonged effusions than those without, and the duration of tube thoracostomy was significantly shorter in patients with collaterals. Embolization of collaterals did not affect the duration of effusions. CONCLUSIONS: Systemic-to-pulmonary arterial collaterals are common after BCPA. In contrast to prior reports, collaterals were not associated with a higher incidence of prolonged effusions after the Fontan procedure in our experience, and did not correlate with poor outcome. PMID- 10800824 TI - Cerebral vascular effects of aortovenous cannulations for pediatric cardiopulmonary bypass. AB - BACKGROUND: The effects of aortovenous cannulations for pediatric cardiopulmonary bypass on cerebral blood flow velocity (CBFV) and electroencephalography (EEG) were evaluated. METHODS: CBFV and EEG were continuously recorded before (baseline), during, and after cannulations until initiation of cooling (mean +/- 95% confidence interval). Vasopressors and/or volume replacement were administered if mean arterial pressure (MAP) decreased below 35 mm Hg. Cannulation-related EEG slowing was used as a criterion for electrocortical alteration. RESULTS: We studied 124 children (3 days to 17 years of age). Aortic and venous cannulations decreased mean CBFV by 10+/-3% and 13+/-4%, respectively, from baseline (p < 0.001). MAP diminished (p < 0.01) by 8+/-3% and 12+/-4%, respectively, from precannulation values (53+/-2 mm Hg). Right atrial cannulation, which was often chosen because the patient was hemodynamically unstable, was more frequently associated with pharmacologic intervention when compared with superior vena cava (SVC) cannulation (p < 0.01). Transient EEG alterations (n = 20) were associated with persistently low MAP (< 30 mm Hg), low CBFV (< 69%), and aortic (n = 4) or SVC (n = 7) cannula malposition. Infants with right atrial cannulation and intervention had more frequent EEG alterations (p = 0.04). Patients requiring intervention were younger (p < 0.01) and had longer hospital stay (p < 0.01) than those without intervention. CONCLUSIONS: Cerebral effects of cannulations are greater in young infants. This was found to be associated with low MAP during heart manipulation or consequence of cannula malpositions. PMID- 10800826 TI - Retrograde cerebral perfusion with hypothermic circulatory arrest in a child. AB - This report describes a 4-year-old boy who presented with infective endocarditis involving the ascending aorta and the arch vessels, with supravalvular aortic stenosis as the underlying pathology. Operation was indicated because of the embolic potential of the vegetations inside the aorta. Retrograde cerebral perfusion was utilized in conjunction with hypothermic circulatory arrest, to flush particulate materials from the arch vessels during operation. PMID- 10800825 TI - Influence of perioperative factors on outcomes in children younger than 18 months after repair of tetralogy of Fallot. AB - BACKGROUND: There has been a trend toward advocating earlier repair of tetralogy of Fallot and avoiding palliative procedures. The impact of this trend on perioperative outcomes has not been adequately documented. METHODS: Data from consecutive patients undergoing repair of tetralogy of Fallot at less than 18 months of age from May 1987 to September 1994 were reviewed. Independent factors associated with duration of stay in the intensive care unit were sought. RESULTS: Repair was performed in 89 infants at a median age of 13 months (range, 15 days to 18 months). A systemic-pulmonary artery shunt was present in 24% of patients. Mean duration of cardiopulmonary bypass was 119+/-37 minutes; 63% of patients received a transannular patch. There were six deaths (7%), all occurring less than 48 hours after repair. The median duration of stay in the intensive care unit was 5 days (range, 1 day to 8 months). Significant independent factors associated with increasing length of intensive care unit stay included younger age at repair, previous shunt, malformation syndrome, increased total dose and number of inotropic agents used, and respiratory complications. Hemodynamic variables serially recorded in the first 48 hours after repair were independently associated with death or prolonged (>7 days) duration of stay. CONCLUSIONS: Although outcomes after repair of tetralogy of Fallot in infants are good, both younger age at repair and previous palliative procedures were associated with longer duration of stay in the intensive care unit. PMID- 10800827 TI - Internal mammary artery perfusing the Leriche's syndrome. AB - Two cases of collateral perfusion of a lower extremity, by way of an internal mammary artery, in the presence of Leriche's syndrome are described. The importance of recognizing this condition prior to coronary artery bypass grafting is emphasized. PMID- 10800828 TI - Postinfarction acute aortic valve regurgitation and cardiac rupture. AB - We report the case of a 71-year-old man who developed acute aortic regurgitation after a myocardial infarct. At operation he was also found to have a contained cardiac rupture. PMID- 10800829 TI - Pulmonary embolectomy in a child. AB - We report the case of a 14-year-old boy treated successfully by pulmonary embolectomy for massive pulmonary embolism, 18 days after bilateral hip surgery. He has a family history of pulmonary embolism and an inherited antithrombin deficiency. His diagnosis was confirmed by spiral computed tomography scan. We believe that pulmonary embolectomy has a role in selected cases in children. PMID- 10800830 TI - Spontaneous closure of a large tracheal fistula due to descending necrotizing mediastinitis. AB - We present a case of a 77-year-old man who had a large tracheal fistula due to descending necrotizing mediastinitis. He underwent long-term care with a respirator after mediastinal drainage operations. The fistula was covered spontaneously with the anterior wall of the esophagus 1.5 months postoperatively. PMID- 10800831 TI - Repair of traumatic aortic valve disruption and descending aortic transection. AB - Traumatic aortic transection after acute deceleration injury remains a highly lethal condition. Concomitant aortic valve disruption is exceedingly rare, and can complicate the timing of surgical management. This report describes the management and outcome of a patient with these types of injuries. PMID- 10800832 TI - Successful myocardial volume reduction in a 9-month-old infant. AB - Left ventricular reduction has shown promise as a treatment for end-stage dilated cardiomyopathy, with restoration of the physiologic ratio between myocardial mass and left ventricular diameter. We present a case of successful partial left ventriculectomy utilizing both lateral and septal wall excision as treatment of dilated cardiomyopathy in a 9-month-old patient. PMID- 10800833 TI - Cervical mediastinoscopy incisional metastasis. AB - Tumor deposit in the cervical mediastinoscopy incision is a rare complication of this operation when it is done to stage lung cancer. The etiology of this complication remains unclear. We present the case of a patient with this condition, discuss the cause and management, and review the literature. PMID- 10800834 TI - Late ventricular structure after partial left ventriculectomy. AB - Nine months after partial ventriculectomy, a 53-year-old man died of progressive heart failure. His heart was examined to determine the alignment of the muscle fibers around the ventricular scar, which was 11 cm long, 1.3 cm thick and 4 cm wide. The scar reached 2 to 12 mm beyond the surgical suture line. The fibers in the middle and subendocardial layers were malaligned, resulting in convergence, compression and regional necrosis. PMID- 10800835 TI - Anomalous pulmonary artery from the aorta via a patent ductus arteriosus: repair in a premature infant. AB - A successful repair of anomalous left pulmonary artery from the ascending aorta via a left ductus arteriosus in a 1 kg baby is reported. Repair was performed at an early age to avoid pulmonary hypertension and left pulmonary artery occlusion. Utilizing the right ductus to perfuse the right lung, surgery was performed without cardiopulmonary bypass. PMID- 10800836 TI - Redo without transfusion in a patient with idiopathic thrombocytopenic purpura. AB - Mitral valve replacement was able to be carried out at redo operation requiring neither allogeneic platelet nor blood transfusion in a patient with idiopathic thrombocytopenic purpura, by means of preoperative high-dose intravenous gamma globulin, autologous blood predonation, use of a centrifugal pump, heparin-coated extracorporeal circuits, and simultaneous splenectomy. PMID- 10800837 TI - Innominate arterial rupture distal to anomalous origin of left carotid artery. AB - Isolated innominate arterial rupture from blunt trauma is rare. We present the case of a pregnant woman with an isolated injury to the innominate artery distal to an anomalous origin of the left common carotid artery. A safe operative technique is described. PMID- 10800838 TI - Konno procedure for infective endocarditis involving aortic valve in a small child. AB - The Konno procedure was performed to replace the aortic valve and the sinus of Valsalva in a 1-year-old child with pneumococcal infective endocarditis. A Dacron (C.R. Bard, Haverhill, PA) graft and a 16 AP ATS Medical valve (ATS Medical Inc, Minneapolis, MN) were used. Adequate debridement could be easily performed. Postoperative recovery was uneventful. The procedure, originally developed for a narrow aortic annulus, could be a good option for the treatment of a small child with aortic valve endocarditis. PMID- 10800839 TI - Massive cerebral embolization: successful treatment with retrograde perfusion. AB - Stroke is an unpredictable and morbid complication of cardiac operations. We report a patient who suffered massive bilateral cerebral embolization during aortic cannulation for coronary bypass. This was treated successfully with hypothermic circulatory arrest and high flow retrograde cerebral perfusion. The patient suffered only minimal neurologic impairment and improved rapidly. She was discharged home on postoperative day 7 neurologically intact. PMID- 10800840 TI - Anatomic biventricular correction by taking down bicaval Glenn shunt. AB - A 7-year-old boy underwent ventricular and atrial septal defect patch closures, pulmonary valvulotomy, left pulmonary artery dilatation, and bicaval Glenn shunt, which maintained good patency between the superior vena cava and right atrium, because of a reduced right ventricular end-diastolic volume (RVEDV). The RVEDV had increased at 15 years of age; therefore, we performed an anatomic biventricular correction by removing the bicaval Glenn shunt. We found that anatomic biventricular correction becomes possible if RVEDV increases postoperatively. PMID- 10800842 TI - Inferior vena caval tumor with intracardiac extension. PMID- 10800841 TI - Switch from a BIVAD to a LVAD in a boy with Kawasaki disease. AB - A 9-year-old boy with Kawasaki disease survived after two severe myocardial infarctions. Thereafter pharmacologically untreatable ventricular arrhythmia and rapidly deteriorating heart failure developed in the patient. After 19 days of biventricular and a further 27 days of left univentricular mechanical circulatory support with the Berlin Heart (Cardiotechnica, Berlin, Germany) assist device the boy successfully underwent heart transplantation. At a follow-up of 54 months, the boy is leading an active and unrestricted life. PMID- 10800843 TI - Minimally invasive resection of congenital subaortic stenosis. AB - Fifteen consecutive patients with membranous subaortic stenosis underwent resection by a minimal-access approach through a partial upper sternotomy using a cardioscope. There were no operative deaths and no postoperative complications. Twelve patients (80%) were extubated in the operating room. The mean hospital stay was 3.1 days, and 3 patients (20%) needed blood products. Our experience demonstrates that this modified approach is a safe and effective surgical option for resection of subaortic stenosis. PMID- 10800844 TI - Mini-sternotomy for off-pump coronary artery bypass grafting. AB - The number of off-pump coronary artery bypass grafting procedures without cardiopulmonary bypass is steadily increasing. We report on a new, minimally invasive surgical approach for off-pump coronary revascularization in multivessel disease. A distal sternotomy is performed to gain access to the left and right internal thoracic arteries and to reach the left anterior descending coronary artery, diagonal branches, and right coronary artery for off-pump revascularization. PMID- 10800846 TI - The heart string: a simple, inexpensive exposure of the heart during coronary artery operations. AB - A method of heart retraction during coronary artery bypass operations is described. The technique improves exposure of the coronary arteries, especially of the circumflex and posterior descending coronary branches during grafting. In addition, it is simple, safe, and inexpensive. Furthermore, this technique can be applied for off-pump coronary artery bypass surgery. PMID- 10800845 TI - Modified pericardial closure technique in patients with ventricular assist device. AB - The use of Gore-Tex (W.L. Gore & Assoc, Flagstaff, AZ) sheet for pericardial closure in patients with left ventricular assist device as a bridge to transplant allows easy dissection of epicardial adhesions and safe resternotomy at the moment of heart transplantation, with no complications or significant cost increase related to the Gore-Tex itself. PMID- 10800847 TI - "Cobrahead" graft for intercostal artery implantation during descending aortic replacement. AB - A technique for a separate sidearm graft ("cobrahead") to facilitate reattachment of intercostal arteries in descending aortic replacement is described. The technique allows for very prompt restoration of spinal cord blood flow (via a Y attachment from the arterial perfusion circuit). The technique permits a simple, quick, and fully accessible anastomosis, technically more facile than the traditional side-to-side anastomosis. None of 7 patients treated with this technique had early or late paraplegia. Preliminary computed tomographic follow up scans confirm patency of the cobrahead graft. PMID- 10800848 TI - Chest trauma during the battle of Troy: ancient warfare and chest trauma. AB - The Iliad is the story of the events related to the siege and battle of Troy presenting an unsurpassed view of the strengths and weaknesses of its main characters. And what a revelation it is to see that the human element that ignites and sustains the flames of war did not change through the centuries and was 3,000 years ago the same as it is today! An added source of fascination in Homer's work is his peculiar description of the injuries suffered by the combatants. That, as well as the presence of physicians in the battlefield, is of special interest for us surgeons. PMID- 10800849 TI - The radial artery: which place in coronary operation? AB - Previous long-term studies have shown unsatisfactory patency of saphenous vein grafts, compared with internal mammary artery grafts. Recently, the use of the radial artery as a coronary artery bypass graft has enjoyed a revival, on the basis of the belief that it will help improving long-term results of coronary operations. The recent report of encouraging 5-year patency rates, supports its continued use as a bypass graft. In this paper, we review the current knowledge about the radial artery as a bypass graft, with special emphasis on the clinical results. PMID- 10800850 TI - Tricuspid valve replacement: results comparing mechanical and biological prostheses. PMID- 10800851 TI - Inflammatory pseudotumors of the lung. PMID- 10800852 TI - Revised pulmonary artery banding. PMID- 10800853 TI - Classification of descending necrotizing mediastinitis. PMID- 10800854 TI - More about obstructive lesions of the right aortic arch. PMID- 10800855 TI - Minimally invasive direct coronary artery bypass controversy: the truly minimally invasive coronary artery bypass versus the "H" graft. PMID- 10800856 TI - Anaphylactic aprotinin reaction. PMID- 10800857 TI - Collagen-thrombin-plasma composite hemostat. PMID- 10800858 TI - Optimal application of gelatin-resorcin-formaldehyde glue with special reference to the quality of mixing. PMID- 10800859 TI - A venous shunt between the innominate vein and pulmonary trunk. PMID- 10800860 TI - Flow measurement in coronary artery surgery. PMID- 10800861 TI - Transatrial cardiac pacing. PMID- 10800862 TI - Posterior approach for plexus tumors in the apical and mediastinal regions. PMID- 10800863 TI - A statewide model detection and prevention program for geriatric alcoholism and alcohol abuse: increased knowledge among service providers. AB - To facilitate the professional development of service providers, the Virginia project on geriatric alcohol abuse and alcoholism developed and used an informational booklet, brochure, and video in a "train the trainer" model. A core group received extensive training, and then trained colleagues in their local communities. Knowledge gains were documented among both trainers and trainees. Follow-up interviews with agency personnel revealed substantial impact on a broad spectrum of service systems and improvements in interagency coordination. Results are discussed in terms of the educational needs of professional service providers regarding the unique aspects of alcoholism and alcohol abuse in the older population. PMID- 10800864 TI - Do consumers who have a choice of treatment have better outcomes? AB - This study used a non-equivalent control group design to investigate the effect of consumer choice of treatment on both process and outcome variables. All study participants suffered from severe mental illness, were homeless at baseline, and were enrolled in a modified Assertive Community Treatment (ACT) program. Consumers in the choice condition had selected the ACT program from a menu of five treatment programs; clients in the no-choice condition were simply assigned to the ACT program by an intake worker. Results found that consumers in the choice condition visited the ACT staff at their offices more than consumers in the no-choice condition, but there were no significant differences between groups on the other treatment process variables. Although consumers in the choice condition increased their income more than consumers in the no-choice condition, there were no significant differences between groups on the other outcome variables (stable housing, psychotic symptoms, depression, and substance abuse). PMID- 10800865 TI - Consumers' and case managers' perceptions of mental health and community support service needs. AB - Consumers with serious and persistent mental illness (N = 385) and their case managers rated the amount of help needed and the amount of help received with mental health and community support services. Consumers also identified their primary source of help with each type of need. Results highlighted areas of agreement and disagreement between consumers' and case managers' perceptions. Consumers' reports revealed a strong reliance on sources of support outside the mental health system (e.g., family and friends) for many community support service needs, interpersonal needs, and crisis-related needs. In general, correlations between consumers' and case managers' ratings of help needed and help received were low. Consumers perceived the majority of their needs to be unmet; case managers perceived the majority of consumer needs to be overly met. Discussion focuses on the importance of increasing consensus between consumers and case managers regarding needs by including consumers in treatment planning and providing them with more information about available services. It is recommended that researchers and evaluators examine perceptions of help needed, help received, and sources of help when assessing service needs. PMID- 10800866 TI - Case management: the current best practices and the next generation of innovation. AB - Eight published literature reviews of case management were evaluated to determine areas of consensus regarding outcomes associated with full service, broker, and hybrid models. Full service was recommended for "best practice" status and the effectiveness of the broker model was seriously questioned. Research evaluating para-professional and peer assisted models of case management, a potential method for improving outcomes, was also reviewed. One promising peer-assisted model was identified and described. Although the use of paraprofessionals and peers in case management holds promise, much more research is needed before the effectiveness of this approach can be confidently determined. PMID- 10800867 TI - Treatment outcome among child psychiatric outpatients in a community mental health center. AB - The study evaluated the effectiveness of mental health center treatments for children aged 5-12 years. The Child Behavior Checklist (CBCL) was administered before and after treatment to the parents of the treatment group (N = 71). The results were analyzed and compared to the results of the pretest and posttest CBCL administered to the parents of a matched group of comparison subjects (N = 33). The results showed that the treatment subjects significantly improved on both the externalizing and internalizing scales of the CBCL. The controls showed no significant change between pretest and posttest scores. As a result, it appears that clinic treatments can help clients with measurable problems decrease problematic behaviors. More data is needed to determine whether clinic treatments such as the ones described in this study can substantially help the most seriously disturbed school-age children. PMID- 10800868 TI - The recovery vision as a service improvement guide for community mental health center providers. AB - Many mental health consumers are calling for Community Mental Health Center services to focus on creating opportunities for recovery. First person accounts of recovery portray a process which entails developing hope, taking personal responsibility for health, and building a broad sense of self that is not illness dominated. Community Mental Health Centers can facilitate recovery by enhancing their clinicians' abilities to (1) communicate hope, (2) teach the skills and knowledge consumers need to effectively manage their health concerns, and (3) support consumers' efforts to get on with life beyond illness. PMID- 10800869 TI - Clinical trials of antiretroviral therapy in developing countries. PMID- 10800870 TI - X chromosome defects and premature ovarian failure. PMID- 10800871 TI - Hendra, Menangle and Nipah viruses. PMID- 10800872 TI - A comparison of two dosing regimens of zidovudine in Thai adults with early symptomatic HIV infection. Conducting clinical HIV trials in South-East Asia. AB - AIM: To compare the clinical and immunological efficacy, and tolerance of two dosage regimens of zidovudine (ZDV) in an adult Thai population with early symptomatic human immunodeficiency virus (HIV) disease and to identify important clinical issues associated with conducting HIV trials in South-East Asia. METHODS: HIV-infected Thai adults, with early symptomatic HIV disease and CD4 lymphocyte counts less than 400/mm3, who were managed in the infectious diseases clinics at two university teaching hospitals in Bangkok, Thailand, were enrolled in a randomised, open-label, dose-regimen comparison trial of ZDV. Two oral ZDV dosing regimens: regimen A, 100 mg tid+200 mg nocte (ZDV-A) vs regimen B, 250 mg bid (ZDV-B) were compared. The main outcome measures were: 1. Clinical efficacy: rate of progression to acquired immunodeficiency syndrome (AIDS) or death. 2. Immunologic efficacy: changes in CD4 lymphocyte numbers compared to baseline; rate of decline of CD4 lymphocyte numbers to less than 100/mm3. 3. Toxicity, as defined by clinical symptomatology and laboratory parameters. RESULTS: Two hundred and four patients were enrolled (103 ZDV-A; 101 ZDV-B) of whom 195 were followed beyond baseline. Patients were typical of those encountered with HIV in Thailand: mean age 33 years; 89% male; 88% heterosexual HIV acquisition; mean baseline CD4 lymphocyte count 241/mm3. Follow-up while on therapy was comparable for the two groups (mean+/-SD): 533+/-236 days (ZDV-A) vs 592+/-210 days (ZDV-B). One hundred and eleven patients (57%; 51 ZDV-A; 60 ZDV-B) were treated for at least 22 months (669+/-30 days). Clinical and immunological outcomes for ZDV-A and ZDV-B, including rate of progression to AIDS or death, development of non AIDS-defining opportunistic infections, mean changes in CD4 lymphocyte numbers/mm3, difference in area under the CD4:time distribution curve and difference in the rate of decline of CD4 lymphocyte numbers to less than 100/mm3, were not significantly different. The presence of oral hairy leukoplakia or unintentional weight loss of 10-20% at enrollment were significantly associated with the later development of AIDS (p=0.03 and 0.04, respectively). ZDV associated toxicity was similar for both regimens. Maintaining protocol adherence and appropriate clinical follow-up emerged as important practical issues. CONCLUSION: In Thai adults, ZDV 100 mg tid+200 mg nocte and ZDV 250 mg bid have similar clinical and immunological efficacy. Rates of ZDV toxicity are comparable to those reported in non-Asian populations. Despite limitations in medical care access and maintaining long-term follow-up, successful trials of antiretroviral agents are feasible in South-East Asia and multi-drug treatment trials should be pursued in appropriate institutions. PMID- 10800873 TI - Calcium intakes among Australian women: Geelong Osteoporosis Study. AB - BACKGROUND: Dietary calcium deficiency may be a risk factor for osteoporosis. AIMS: To estimate habitual calcium intakes and prevalence of calcium supplementation among free-living Australian women and validate a calcium specific food-frequency questionnaire. METHODS: Calcium intakes for 1045 randomly selected women (20-92 years) were estimated by questionnaire which was tested against estimates from four day weighed records kept by 32 randomly selected women. RESULTS: The mean difference between calcium estimates was not statistically significantly different from zero (mean difference=121 mg; standard deviation of differences=357 mg; p>0.05). There was moderate agreement (weighted kappa=0.4) between methods in ranking subjects into tertiles of calcium intake. Mean dietary calcium intakes were 615 mg/day for 20-54 years, 646 mg/day for 55 92 years and 782 mg/day for lactating women. Seventy-six per cent of women aged 20-54 years, 87% of older and 82% of lactating women had intakes below the recommended dietary intake (RDI). There was no association detected between calcium intake and age. Dairy foods provided 79.0% of dietary calcium intake. Calcium supplements were used by 6.6% and multivitamins by a further 4.3% of women. Supplementation was independent of dietary calcium intake and more likely used by postmenopausal women. CONCLUSIONS: Our results suggest that 76% of women consume less than the RDI even when supplemental calcium is included. Furthermore, 14% have less than the minimal requirement of 300 mg/day and would, therefore, be in negative calcium balance and at risk of bone loss. Despite advertising campaigns promoting better nutrition and increased awareness of osteoporosis, many women are failing to achieve an adequate calcium intake. PMID- 10800874 TI - HLA-B27 expression and reactive arthritis susceptibility in two patient cohorts infected with Salmonella Typhimurium. AB - BACKGROUND: Reactive arthritis (ReA) is an inflammatory arthritis triggered by certain gastrointestinal and genitourinary infections. Single source outbreaks of triggering infections provide an opportunity to elucidate host susceptibility factors in this disease. AIM: To determine the role of Major Histocompatibility Complex (MHC) Class I alleles in ReA susceptibility after two large single source outbreaks of Salmonella Typhimurium gastroenteritis. METHODS: A questionnaire screening for features of ReA and a request for HLA class I typing were sent to all patients affected by two single source outbreaks of S. Typhimurium gastroenteritis. Individuals with arthritis of recent onset were interviewed, examined and diagnostic criteria for ReA applied. RESULTS: Nineteen cases of reactive arthritis, 11 female, were diagnosed in the 424 respondents with S. Typhimurium gastroenteritis from both outbreaks. Clinical features of the arthritis were similar to those described after other large single source outbreaks of Salmonella infection. HLA-B27 was expressed by only two of the 19 ReA patients and therefore did not predict susceptibility to this form of arthritis. Caucasians were, however, more likely to develop reactive arthritis than Asians. CONCLUSIONS: In this study, susceptibility to ReA was not increased in HLA-B27 positive individuals or males but was greater in those of Caucasian descent. PMID- 10800875 TI - Teaching asthma management: an evidence-based educational approach. AB - BACKGROUND: Asthma management plans and asthma education have been incorporated into recent guidelines, and implementation of these reduces asthma morbidity. Junior doctors are frequently involved in asthma management, but teaching about asthma in medical school is seldom assessed. We studied asthma knowledge, confidence in practical asthma management, and the effectiveness of an interactive asthma education workshop in final year medical students. Thirty-five students attended the workshop, with 26 returning questionnaires on both of the two sessions. METHODS: Students attended a two hour asthma workshop comprising interactive teaching sessions on management of acute and stable asthma, and a practical session using asthma devices. Theoretical knowledge was assessed by a 20 item questionnaire, completed immediately prior to the workshop and after two weeks, at a feedback session. Practical knowledge was assessed by confidence scores in use of asthma devices on a five point scale (not at all confident to very confident), before, immediately after the workshop, and after two weeks. RESULTS: Students scored poorly in questions on: predictors of asthma mortality, nebuliser and Turbuhaler use, asthma management plans, and physical signs in acute asthma. Mean (SE) knowledge score increased significantly from 64.4% (6.1) to 74.2% (5.6) (p<0.05; paired t-test) at two weeks. Confidence scores rose significantly for all practical aspects of asthma management (p<0.01; Wilcoxon matched pairs test), and declined at two weeks, but still remained significantly higher than at baseline (p<0.05). CONCLUSIONS: Medical students had poor knowledge about several important features of asthma care. The asthma workshop was effective in increasing knowledge and confidence in management of asthma in the short term, and could be useful in optimising implementation of asthma guidelines in clinical practice and potentially later in training of junior medical officers. PMID- 10800876 TI - Does high dose ipratropium bromide added to salbutamol improve pulmonary function for patients with chronic obstructive airways disease in the emergency department? AB - BACKGROUND: To determine the effect of high dose ipratropium bromide, both alone or in combination with standard dose salbutamol, on pulmonary function in patients presenting to the Emergency Department (ED) with acute exacerbation of chronic obstructive airways disease (COAD). METHOD: The trial was a prospective, randomised, double blind trial of adult patients with COAD. All patients received nebulised salbutamol 5 mg and 500 microg ipratropium and hydrocortisone 250 mg i.v. at time=0, then were randomised to receive further nebulisers at time=15 minutes and time=30 minutes of salbutamol 5 mg combined with ipratropium 500 microg or salbutamol 5 mg alone or ipratropium 500 microg alone. Pulmonary function tests were conducted at time=0 and time=90 minutes. The primary endpoints were absolute and percent change in FEV1. RESULTS: The group randomised to receive 5 mg salbutamol and 500 microg ipratropium (n=18) showed a mean percentage change of FEV1 of 6.4% with a mean absolute change of 0.06 L (SD 0.18 L). Those who received 5 mg salbutamol (n=16) had a mean percentage change of 18.6% with a mean absolute change of 0.13 L (SD of 0.21 L). Those who received 500 microg ipratropium (n=16) had a mean percentage change of 4.8% with a mean absolute change of 0.023 L (SD of 0.07 L). There was no significant difference between the groups in FEV1 (p=0.56 for percentage change; p=0.36 for absolute change). CONCLUSION: The addition of 500 microg ipratropium to 5 mg salbutamol in subsequent nebulisers adds no benefit to pulmonary function after the initial nebuliser of both bronchodilators in the treatment of COAD in the ED. PMID- 10800877 TI - Fasting plasma glucose in non-diabetic elderly women predicts increased all causes mortality and coronary heart disease risk. AB - AIM: To confirm the hypothesis that upper normal plasma glucose levels in non diabetic subjects are independently predictive of mortality and cardiovascular disease (CVD). METHODS: The study reports on 113 months' follow-up in a prospective study of CVD in the Australian elderly, The Dubbo Study. The cohort, first examined in 1988-89, consisted of 2805 men and women 60 years and older. Of the cohort, 2419 (86%) were defined as non-diabetic. The prediction of outcomes by quartile of fasting plasma glucose was examined in a Cox proportional hazards model, after linkage to hospital and death records. RESULTS: All-causes mortality increased progressively across quartile of fasting plasma glucose in both sexes, reaching statistical significance only in women. Coronary heart disease (CHD) incidence increased similarly, the increases being proportionately greater in women. Ischaemic stroke did not show a consistent gradient with fasting plasma glucose. After adjustment for age and other risk factors, all-causes mortality, CHD and ischaemic stroke incidence were not significantly related to plasma glucose in men. In women, all-causes mortality and CHD incidence showed a significant gradient with glucose quartile. Hazard Ratio (95% confidence intervals) for death in glucose Quartile IV (5.3-6.0 mmol/L) was 1.49 (1.03-2.14) and for CHD incidence was 1.52 (1.08-2.15). Subjects in the upper quartiles of fasting plasma glucose showed a clustering of overweight, hypertension, elevated serum triglycerides, reduced high density lipoprotein cholesterol and excess of small dense low density lipoprotein, suggestive of the Insulin Resistance Syndrome. CONCLUSION: Fasting plasma glucose levels in the upper normal range in non-diabetic elderly subjects appear to be associated with increased all-causes mortality and CHD, especially in women. PMID- 10800879 TI - Intravenous magnesium for the treatment of cardiac arrhythmias. PMID- 10800878 TI - The angiotensin converting enzyme (ACE) inhibitor, perindopril, modifies the clinical features of Parkinson's disease. AB - BACKGROUND: Animal studies have demonstrated an interaction within the striatum between the angiotensin and dopaminergic systems. In rats, the angiotensin converting enzyme (ACE) inhibitor, perindopril, crosses the blood brain barrier and increases striatal dopamine synthesis and release. In humans, angiotensin type 1 receptors have been found on dopaminergic neurons in the substantia nigra and striatum. In Parkinson's disease, there is a marked reduction of these receptors associated with the nigrostriatal dopaminergic neuron loss. AIMS: We performed a double blind placebo controlled crossover pilot study in seven patients to investigate the effect of the ACE inhibitor, perindopril on the clinical features of moderately severe Parkinson's disease. RESULTS: After a four week treatment period with perindopril, patients had a faster onset in their motor response to L-dopa and a reduction in 'on phase' peak dyskinesia, p=0.021 and p=0.014 respectively. Patients also reported more 'on' periods during their waking day in their movement diary, p=0.007. Perindopril was well tolerated without any significant postural hypotension or renal dysfunction. CONCLUSIONS: These results suggest that ACE inhibitors such as perindopril may have a place in the management of motor fluctuations and dyskinesia in Parkinson's disease and justify further study. PMID- 10800880 TI - Simplistic physical activity guidelines: the need for trials. PMID- 10800881 TI - The dawning of a new era for physical inactivity as a health risk factor. PMID- 10800882 TI - The clinician-scientist in the 21st century. PMID- 10800883 TI - Economic rationalism in health and education; impact on the academic physician. PMID- 10800884 TI - The quandary of policy on illicit drugs. PMID- 10800885 TI - Health policy--taking physicians' views to Canberra. PMID- 10800886 TI - The neurology of George Edward Rennie (1861-1923). PMID- 10800887 TI - Early menopause in a family carrying a fragile X premutation. PMID- 10800888 TI - De novo deletion of Xq associated with premature ovarian failure. PMID- 10800889 TI - Progression to accelerated hypertension in untreated primary aldosteronism. PMID- 10800890 TI - Left main coronary artery disease in a 40-year-old man receiving HIV protease inhibitors. PMID- 10800891 TI - Prostatic abscess: another manifestation of tuberculosis in HIV-infected patients. PMID- 10800892 TI - A case of quinine-induced thrombocytopenia and neutropenia. PMID- 10800893 TI - Treatment of peritoneal dialysis related peritonitis. PMID- 10800894 TI - A case of diffuse panbronchiolitis. PMID- 10800895 TI - Geriatric evaluation and management in a New Zealand hospital. PMID- 10800896 TI - Cryoglobulinaemia causing systemic vasculitis and interstitial lung disease. PMID- 10800897 TI - Early recurrence of type 1 membranoproliferative glomerulonephritis following cadaveric renal transplantation. PMID- 10800898 TI - Successful use of dapsone in refractory pregnancy-associated idiopathic thrombocytopenic purpura. PMID- 10800899 TI - Clarity in professional writing. PMID- 10800900 TI - Nasal profile changes after maxillary impaction and advancement surgery. AB - PURPOSE: The profile changes in the nasal tip and columello-labial region that occur after maxillary advancement and impaction surgery were measured to test the hypothesis that a subspinal osteotomy would induce less nasal tip change and would result in a more acute columello-labial angle than a conventional Le Fort I type osteotomy. PATIENTS AND METHODS: The lateral cephalograms of 2 matched groups of 23 advancement/impaction cases with and without subspinal osteotomy were analyzed electronically. RESULTS: There was no difference in nasal tip elevation and change in nasal tip projection between the 2 groups. The columello labial angle was, on average, unchanged by the surgery. A linear correlation with a weak clinical relevance could be demonstrated between nasal tip projection and maxillary advancement in the group that was treated without subspinal osteotomy. Such a correlation was not detected for nasal tip elevation in either of the groups. Palatal plane rotation had a significant influence on nasal tip projection but not on tip elevation in both groups. The correlation was strongest in the group treated conventionally. The multiple regression equation with maxillary advancement and rotation as predictors had a r2 of .6071 (nasal tip projection = 0.9 + 0.19 maxillary advancement + 0.18 palatal plane inclination) in this group. CONCLUSION: The results indicate that the advancing piriform aperture pushing on the alae, and not the nasal spine, is responsible for the increase in nasal tip projection. The subspinal osteotomy is not superior to the conventional Le Fort I-type osteotomy in regard to minimizing nasal tip changes and obtaining control over the columello-labial angle. PMID- 10800901 TI - Diagnosis and treatment of cervical tuberculous lymphadenitis. AB - PURPOSE: This study presents the long-term results of treatment of a series of patients with tuberculous mycobacterial lymphadenitis of the head and neck. PATIENTS: Twenty-one patients were seen in a 10-year period. The median age at onset was of 41.2 years (range, 4 to 79 years), and the male-to-female ratio was 11:10. Sixteen patients were of Ethiopian origin, 3 from the former USSR, and 2 were Israeli women (1 of Indian and 1 of Morrocan origin). Symptoms started between 2 weeks and 6 months before presentation (mean, 5.8 weeks). Most patients had negative chest radiographs, a variable response to the tuberculin skin test, and a negative culture for mycobacterial organisms. RESULTS: Fine-needle aspiration (FNA) of the cervical lymph nodes was the most reliable method to confirm the bacteriologic agent causing the lymphadenopathy. Acid-fast bacilli smears of the aspirate were positive in all but 3 patients, whereas histologic examination of the lymph nodes gave diagnostic results in only two thirds of cases examined. All patients were treated with antituberculous chemotherapy. Sixteen patients also underwent surgical excision of their cervical lymph nodes, and all of them showed a complete response to the combined treatment. The remaining patients reacted to chemotherapy alone with complete cure. One patient died of gastric carcinoma, and the only acquired immune deficiency syndrome (AIDS) patient died a year later of cytomegalovirus encephalitis. CONCLUSION: The most reliable indicator of cervical mycobacterial infection is an acid-fast smear from the FNA specimen. Antituberculous chemotherapy, with or without surgical excision of the involved cervical lymph nodes, is the method of choice for treatment of this disease. PMID- 10800902 TI - The 2-piece Le Fort I osteotomy for cranial base access: an evaluation of 9 patients. AB - PURPOSE: This article discusses the use of the 2-piece Le Fort I osteotomy to gain access to the cranial base and the authors' experiences with this technique. METHODS: Medical records of patients who required surgical access to the cranial base were retrospectively reviewed, and demographic data, procedures done, postoperative results, and complications were recorded. RESULTS: Nine patients underwent a 2-piece Le Fort I osteotomy. Complications included development of a pseudoaneurysm of the internal carotid artery in 1 patient, 2 cerebrospinal fluid leaks, both of which resolved, and 1 patient who lost 2 teeth and some associated bone. CONCLUSION: The 2-piece Le Fort I osteotomy provides excellent access to the clival region of the cranial base with minimal complications. PMID- 10800903 TI - Magnetic resonance imaging findings and clinical symptoms in the temporomandibular joint in patients with mandibular continuity defects. AB - PURPOSE: This study explored the morphologic changes and clinical symptoms related to the temporomandibular joint (TMJ) associated with long-term lack of mandibular continuity. PATIENTS AND METHODS: The subjects were 20 patients (40 TMJs) who underwent mandibulectomy including the condyle or segmental mandibulectomy without mandibular reconstruction more than 6 months previous to the study. The 40 TMJs were classified into the following 3 groups: group I: TMJs with a major mandibular fragment including the mandibular body (n = 21), group II: TMJs with a minor mandibular fragment including only the condylar process or the mandibular ramus (n = 10), and group III: TMJs without a condyle (n = 9). All TMJs were examined with respect to disc position, condylar position, bony changes, and appearance of joint effusion on magnetic resonance imaging, as well as for the presence of joint or muscle pain and joint noise. RESULTS: The rate of anterior disc displacement was 28.6% in group I, 10.0% in group II, and 100% in group III. All condyles in group I were located in the glenoid fossa. Seven of 10 condyles in group II were dislocated anteriorly out of the glenoid fossa. However, these condyles were situated in the intermediate zone of the disc. Osteoarthritic changes was found in 1 joint in group II. Joint effusion was not observed in group I. However, joint effusion was detected in the posterior region of the upper joint compartment in 7 of 10 TMJs in group H and in 8 of 9 TMJs in group III. In group II, the 7 TMJs with joint effusion were the same joints with anteriorly dislocated condyles. Clinical symptoms of TMJ pain, TMJ noise, or muscular pain were not found in any patients. CONCLUSIONS: Partial mandibulectomy does not affect the relationship between the disc and the condyle or the bony condition of the TMJ. The disc without a condyle will become anteriorly displaced with time. The condyle not connected to the mandibular body often dislocates anteriorly out of the glenoid fossa, while retaining a normal relationship with the disc. Joint effusion, which is detected in nonfunctional TMJs, is not associated with an inflammatory reaction in the joint compartment. PMID- 10800904 TI - Morphologic changes in forearm flaps of the oral cavity. AB - PURPOSE: This study was undertaken to investigate the morphologic changes in the forearm flap in the oral environment. PATIENTS AND METHODS: Histologic evaluation of biopsy specimens from 20 forearm flaps was done 1 to 75 months after intraoral reconstruction after treatment of cancer. Hematoxylin and eosin and Azan were used to stain the deparaffinized sections. RESULTS: The clinical features of the grafted flap seemed to depend on the time since the operation. The cornified layer of the epidermis of the flaps showed thinning histologically. CONCLUSION: After 11 months of exposure to an intraoral environment, "mucosalization" of the forearm flap is evident. PMID- 10800905 TI - Complications of temporomandibular joint arthroscopy: a retrospective analysis of 301 lysis and lavage procedures performed using the triangulation technique. AB - PURPOSE: This study evaluated the complications of temporomandibular joint (TMJ) arthroscopic lysis and lavage performed using the triangulation technique. PATIENTS AND METHODS: The 202 consecutive patients (301 joints) who underwent arthroscopic lysis and lavage between 1992 and 1995 were retrospectively surveyed. The preoperative diagnosis of all patients was anterior disc displacement without reduction. RESULTS: Complications were observed in 31 (10.3%) of 301 cases of arthroscopic lysis and lavage. Of these, otologic complications were found in 26 cases (8.6%): blood clots in the external auditory canal in 9; laceration of external auditory canal in 7; partial hearing loss in 5; ear fullness in 2; vertigo in 1; and perforation of tympanic membrane with laceration of external auditory canal in 1. Neurologic injuries were found in 5 cases (1.7%): fifth cranial nerve injuries in 3; and seventh cranial nerve injuries in 2. No other complications were observed. CONCLUSIONS: A high level of understanding of the regional anatomy helps reduce the complications associated with arthroscopic lysis and lavage in the TMJ. PMID- 10800906 TI - Effects of latency and rate on bone formation in a porcine mandibular distraction model. AB - PURPOSE: Long treatment protocols currently limit the application of distraction osteogenesis (DO). The purpose of this study was to develop a porcine model for DO and to define the effects of latency and distraction rate on bone formation. MATERIALS AND METHODS: Distractors were placed through submandibular incisions. For analysis of latency, mandibular osteotomies were distracted after 0 (n = 3) or 4 (n = 2) days at a rate of 1 mm/d (7 days) with 14 days fixation. For analysis of rate, osteotomies were distracted at 1 mm (n = 4), 2 mm (n = 4), or 4 mm (n = 4) per day to produce a 12-mm gap with 24 days fixation. DO wounds were assessed in vivo with bimanual palpation to detect mobility across the gap. Harvested specimens were evaluated by gross appearance and bimanual palpation. Standardized radiographic techniques were used to estimate bone density. RESULTS: During DO, animals showed normal activity. There were no infections, and no distractors were removed prematurely. Clinical and radiographic evaluation of the groups that underwent distraction after 0- or 4-day latency showed equivalent healing. In the comparison of distraction rates, stability was greatest in the group distracted at 1 mm/d. CONCLUSIONS: A porcine model for mandibular DO has been developed in which mandibular lengthening was successfully performed without latency and at a rate of 1 mm/d. The relationships among latency, gap size, rate, and duration of fixation are poorly understood and can be defined with relevant animal models. PMID- 10800907 TI - Morphologic and biomechanical correlates with maximum bite forces in orthognathic surgery patients. AB - PURPOSE: The purpose of this study was to determine which factors of craniofacial morphology best predict maximum bite forces and jaw muscle strength (based on [electromyogram] EMG/force slopes) in patients selected for various orthognathic surgical procedures. These factors were then compared for their ability to separate orthognathic surgery patients by their clinical diagnosis. PATIENTS AND METHODS: Standard lateral cephalograms were taken of 121 orthognathic surgery patients (before treatment) and 80 control subjects to establish multivariate sagittal and biomechanical factors of craniofacial form. Maximum and submaximal bite forces were recorded at 8 tooth positions for each subject. EMG activity was recorded for 3 pairs of muscles (anterior temporalis, posterior temporalis, and superficial masseter) during the isometric bites. The EMG and bite force measurements were used to calculate EMG/force slopes as a measure of jaw muscle strength. The study looked for significant correlations between the morphologic factors and maximum bite force or jaw muscle strength. RESULTS: Factor analysis determined 12 sagittal and 6 biomechanical factors. However, only 3 of the sagittal and 4 of the biomechanical factors were significantly correlated with maximum bite force or jaw muscle strength. Factors reflecting jaw size were correlated with maximum bite forces and jaw muscle strength but generally did not separate patient groups. The factor most strongly correlated with maximum bite forces separated patients by their relative difference between anterior and posterior facial height. The factor for anteroposterior maxillomandibular discrepancies was not correlated with maximum bite force or jaw muscle strength. CONCLUSIONS: Many cephalometric measurements used to diagnose craniofacial deformities and to assign patients to particular orthognathic surgical procedures are not correlated with maximum bite forces or jaw muscle strength. Only measurements reflecting relative differences between anterior and posterior facial height are both strongly correlated with maximum bite force and reflect assignment of surgical procedures. PMID- 10800908 TI - Tumor necrosis factor-alpha in synovial fluid and plasma from patients with chronic connective tissue disease and its relation to temporomandibular joint pain. AB - PURPOSE: The purpose of this study was to determine the level of tumor necrosis factor-alpha (TNF-alpha) in the temporomandibular joint (TMJ) synovial fluid (SF TNF-alpha) and blood plasma (P-TNF-alpha) of patients with chronic inflammatory connective tissue disease and investigate its relation to TMJ pain, hyperalgesia, and allodynia. PATIENTS AND METHODS: Twenty-four patients with a diagnosis of chronic inflammatory connective tissue disease and TMJ pain were included in the study. Visual analog scale, tenderness of the TMJ, and pain at mandibular movements were registered, and the pressure pain threshold and pressure pain tolerance levels were measured. TMJ synovial fluid samples and blood plasma were analyzed for TNF-alpha and the levels related to TMJ pain, hyperalgesia, and allodynia. RESULTS: TNF-alpha was present in the TMJ synovial fluid of 8 of 24 patients at levels significantly exceeding those in plasma at the same visit. The presence of SF-TNF-alpha showed a significant positive correlation to TMJ pain at maximum voluntary mouth opening and tenderness to posterior palpation of the TMJ. CONCLUSION: Local production of TNF-alpha occurs in the TMJ synovium of patients with chronic inflammatory connective tissue disease. Pain on mandibular movement and tenderness on posterior palpation (allodynia) of the TMJ is related to the level of SF-TNF-alpha. PMID- 10800909 TI - Dental extraction wound management: medicating postextraction sockets. AB - The rather impressive percentage of extraction sites undergoing clot loss and deranged healing results in significant morbidity for the patient and frequent visits to the surgeon to effect relief of discomfort, most often by the use of anodyne dressings. The amount of work lost by patients needing such palliative treatment, and loss of productive time for the surgeon, translate into an unknown, but potentially large, economic loss to society. This would mandate that economical methods of ensuring normal extraction socket healing with minimal morbidity be developed. The most useful socket medicaments to prevent socket healing derangements would include broad-spectrum antibiotics, specifically clindamycin and tetracycline. Not discussed in this article, but possibly germane to the subject of clot stabilization and healing, is consideration of resorbable substances such as gelatin sponge, polylactic acid, and methylcellulose as clot stabilizing socket implants. The record of such substances in preventing AO is mixed, but the combinations of these inexpensive materials with topical socket medicaments may yield a decreased tendency for clot lysis and greater mechanical strength to the bulk blood clot, as has been demonstrated with difficult mandibular third molar impactions in one study involving polylactic acid, tetracycline, and hydrocortisone. PMID- 10800910 TI - Dental extraction wound management: a case against medicating postextraction sockets. PMID- 10800911 TI - Rapidly growing mass in the parotid gland. PMID- 10800912 TI - Mandibular fracture resulting in displacement of the inferior alveolar nerve and allodynia. PMID- 10800913 TI - Eosinophilic granuloma of the mandibular condyle. PMID- 10800914 TI - Postextraction panfacial cellulitis (Sweet's syndrome) mimicking an odontogenic infection. PMID- 10800915 TI - Parathyroid carcinoma diagnosed on the basis of a giant cell lesion of the maxilla. PMID- 10800916 TI - Giant pleomorphic adenoma of the lacrimal gland. PMID- 10800917 TI - Mycobacterium abscessus infection after facelift surgery: a case report. PMID- 10800918 TI - Removal of a premolar with extreme distal migration by sagittal osteotomy of the mandibular ramus: report of case. PMID- 10800919 TI - Parafunctional habits causing temporomandibular disorders. PMID- 10800920 TI - Cysteine-string protein: the chaperone at the synapse. AB - Cysteine-string protein (Csp) is a major synaptic vesicle and secretory granule protein first discovered in Drosophila and Torpedo. Csps were subsequently identified from Xenopus, Caenorhabditis elegans, and mammalian species. It is clear from the study of a null mutant in Drosophila that Csp is required for viability of the organism and that it has a key role in neurotransmitter release. In addition, other studies have directly implicated Csp in regulated exocytosis in mammalian neuroendocrine and endocrine cell types, and its distribution suggests a general role in regulated exocytosis. An early hypothesis was that Csp functioned in the control of voltage-gated Ca2+ channels. Csp, however, must have an additional function as a direct regulator of the exocytotic machinery as changes in Csp expression modify the extent of exocytosis triggered directly by Ca2+ in permeabilised cells. Csps possess a cysteine-string domain that is highly palmitoylated and confers membrane targeting. In addition, Csps have a conserved "J" domain that mediates binding to an activation of the Hsp70/ Hsc70 chaperone ATPases. This and other evidence implicate Csps as molecular chaperones in the synapse that are likely to control the correct conformational folding of one or more components of the vesicular exocytotic machinery. Targets for Csp include the vesicle protein VAMP/synaptobrevin and the plasma membrane protein syntaxin 1, the significance of which is discussed in possible models to account for current knowledge of Csp function. PMID- 10800921 TI - Effect of p-chloroamphetamine on 5-HT1A and 5-HT7 serotonin receptor expression in rat brain. AB - The aim of this study was to investigate if p-chloroamphetamine (PCA), which is neurotoxic to serotonin (5-HT) nerve terminals, was able to induce, like 3,4 methylenedioxymethamphetamine, a region-specific regulation of 5-HT1A receptor mRNA expression. The effect of PCA on the expression of 5-HT7 receptors, which share some pharmacological properties with 5-HT1A receptors, was comparatively studied. PCA (2 x 5 mg/kg) produced a lasting depletion of 5-HT content in the rat frontal cortex and hippocampus. In the hippocampus, the maximal 5-HT depletion was found on day 21 (-70%), whereas in the cortex, the highest 5-HT depletion was found on day 14 (-73%), with a partial but significant recovery on day 21. At the latter time point, 5-HT1A receptor mRNA expression was increased by 80% in the cortex and decreased by 50% in the hippocampus. The 5-HT1A receptor mRNA expression was also enhanced after exposure to PCA of rat cortical but not of hippocampal primary cultures. In regard to 5-HT7 receptor mRNA expression, the most remarkable change after PCA was the great increase (+200%) in the brain stem. Binding studies to 5-HT1A receptors matched the changes in receptor mRNA expression. Gel shift assays revealed enhanced nuclear protein binding to the KB sequence with use of cortical but not hippocampal extracts of PCA-treated rats. Overall, the data show region-specific changes in 5-HT receptor-type expression that may not be entirely dependent on the neurotoxic effect of PCA on 5-HT terminals. PMID- 10800922 TI - Characterization in cultured cerebellar granule cells and in the developing rat brain of mRNA variants for the NMDA receptor 2C subunit. AB - N-Methyl-D-aspartate (NMDA) receptors are heteromeric structures resulting from the association of at least two distantly related subunit types, NR1 and one of the four NR2 subunits (NR2A-NR2D). When associated with NR1, the NR2 subunits impose specific properties to the reconstituted NMDA receptors. Although the NR1 mRNAs are expressed in the majority of central neurons, the NR2 subunits display distinct patterns of expression in the developing and adult rat brain. The NR2C subunit is barely expressed in the rat forebrain, whereas its expression increases substantially in the granule cells in the course of cerebellar development. We have identified novel NR2C splice variants in cultured cerebellar granule cells as well as in the developing cerebellum. When compared with the prototypic NR2C mRNA, these variants carry one (NR2Cb) or two (NR2Cd) insertions or a deletion (NR2Cc) and encode putative NR2C polypeptides that terminate between the third and fourth membrane segments or between the first and second membrane segments. RT-PCR analysis and in situ hybridization show that expression of the splice variants is developmentally regulated, both in the cerebellum and in the hippocampus. Electrophysiological recordings and microfluorimetry emissions in transfected human embryonic kidney 293 cells indicate that the NR2Cb variant, when expressed in combination with NR1, does not contribute to the formation of functional receptor channels. The significance of theses findings is discussed. PMID- 10800923 TI - Identification of variants and dual promoters of murine serine/threonine kinase KKIAMRE. AB - KKIAMRE is a serine/threonine protein kinase whose transcripts increase in the deep cerebellar nuclei of the rabbit after eyeblink conditioning, a model of associative learning and memory. We here characterized the expression, isoforms, and promoters of murine KKIAMRE gene. The expression of KKIAMRE was detected, by in situ hybridization and immunohistochemistry, in neurons in various brain regions including deep cerebellar nuclei. The gene spans approximately 40 kb and consists of 15 exons. Analysis of cDNA clones revealed multiple variants, having diversity in the putative carboxy-terminal regulatory domain, generated by alternative splicing and intraexonal termination. Furthermore, they had alternative 5' noncoding sequences. Primer extension, RNase protection, and transient expression assays revealed that two alternative promoters linked to distinct noncoding exons direct the expression of KKIAMRE. The gene was mapped on chromosomes 5 and 4 in mouse and human, respectively. PMID- 10800924 TI - Efficient gene transfer and expression of biologically active glial cell line derived neurotrophic factor in rat motoneurons transduced wit lentiviral vectors. AB - Several studies have shown the ability of human immunodeficiency virus type 1 (HIV1)-based lentiviral vectors to infect nondividing brain and retinal neurons with high efficiency and long-term expression of the transduced gene. We show that purified embryonic motoneurons can be efficiently (>95%) transduced in culture using an HIV1-based lentiviral vector encoding LacZ. Expression of beta galactosidase was observed for at least 9 days in these conditions. Furthermore, motoneurons transduced with a lentiviral vector expressing glial cell line derived neurotrophic factor survived in the absence of additional trophic support, showing that the overexpressed protein was biologically active. Our results demonstrate the potential of lentiviral vectors in studying the biological effects of proteins expressed in motoneurons and in the development of future gene therapy for motoneuron diseases. PMID- 10800925 TI - A response element for the homeodomain transcription factor Ptx3 in the tyrosine hydroxylase gene promoter. AB - Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the biosynthesis of catecholamines, which takes place in different types of neuronal systems and nonneuronal tissues. The transcriptional regulation of the TH gene, which is complex and highly variable among different tissues, reflects this heterogeneity. We recently isolated a homeodomain transcription factor, named Ptx3, that is uniquely expressed in the dopaminergic neurons of the substantia nigra pars compacta and ventral tegmental area, which together form the mesencephalic dopaminergic system. This strict localization and its coinciding induction of expression with the TH gene during development suggested a possible role for this transcription factor in the control of the TH gene. We report here the presence of a responsive element for Ptx3 located at position -50 to -45 of the rat TH promoter. Transient transfections using TH promoter constructs and electrophoretic mobility shift assays using Ptx3-containing nuclear extracts demonstrated that this region binds Ptx3 protein and confers a transcriptional effect on the TH gene. Depending on the cell type, the effect of Ptx3 was an eight- to 12-fold enhancement of TH promoter activity in Neuro2A neuroblastoma cells, or a 60-80% repression in nonneuronal human embryonic kidney 293 cells. Despite the close association of the Ptx3-binding site and the major cyclic AMP response element in the TH gene, no interplay was found between Ptx3 and cyclic AMP-modulating agents. In combination with the orphan nuclear receptor Nurr1, which is required for the induction of the TH gene in mesencephalic dopaminergic neurons, the TH promoter activity to Ptx3 was enhanced in Neuro2A cells. Nurr1 alone displayed only very weak activity on the TH promoter in this cell type. The results demonstrate that the homeodomain protein Ptx3 has the potential to act on the promoter of the TH gene in a markedly cell type-dependent fashion. This suggests that Ptx3 contributes to the regulation of TH expression in mesencephalic dopaminergic neurons. PMID- 10800926 TI - rTLE3, a newly identified transducin-like enhancer of split, is induced by depolarization in brain. AB - The transducin-like enhancers of split are a family of mammalian proteins that share sequence homology with the Drosophila protein Groucho. Using representational difference analysis, we isolated the cDNA for a previously unidentified gene, rTLE3 (rat transducin-like enhancer of split 3), as a sequence induced by depolarization and forskolin, but not by neurotrophins or growth factors, in PC12 pheochromocytoma cells. rTLE3 encodes the protein rTLE3, a 764 amino acid orthologue of mouse and human TLE3. R-esp2, the gene encoding the closest related rat protein, is not induced by any of the four treatments in PC12 cells. rTLE3 and R-esp2 have different patterns of expression in the adult rat CNS and other tissues. After systemic administration of kainic acid, rTLE3 is induced specifically in the dentate gyrus of the hippocampus. We propose that members of the transducin-like enhancer of split family of proteins may have distinct functions in the mature CNS, in addition to their functions during development. PMID- 10800927 TI - Functional glucokinase isoforms are expressed in rat brain. AB - Recently, the description of glucokinase mRNA in certain neuroendocrine cells has opened new ways to characterize this enzyme in the rat brain. In this study, we found glucokinase mRNA and a similar RNA splicing pattern of the glucokinase gene product in rat hypothalamus and pancreatic islets; the mRNA that codes for B1 isoform was the most abundant, with minor amounts of those coding for the B2, P1, P2, P1/B2, and P2/B2 isoforms. Glucokinase gene expression in rat brain gave rise to a protein of 52 kDa with a high apparent Km for glucose and no product inhibition by glucose 6-phosphate, with a contribution to the total glucose phosphorylating activity of between 40 and 14%; the hypothalamus and cerebral cortex were the regions of maximal activity. Low and high Km hexokinases were characterized by several criteria. Also, using RT-PCR analysis we found a glucokinase regulatory protein mRNA similar to that previously reported in liver. These findings indicate that the glucokinase present in rat brain should facilitate the adaptation of this organ to fluctuations in blood glucose concentrations, and the expression of glucokinase and GLUT-2 in the same hypothalamic neurons suggests a role in glucose sensing. PMID- 10800928 TI - A minimal promoter for the GABA(A) receptor alpha6-subunit gene controls tissue specificity. AB - The ability of nerve cells to regulate the expression of specific neurotransmitter receptors is of central importance to nervous system function, but little is known about the DNA elements that mediate neuron specific gene expression. The type A gamma-aminobutyric acid (GABA(A)) receptor alpha6-subunit gene, which is expressed exclusively in cerebellar granule cells, presents a unique opportunity to study the cis elements involved in restricting gene expression to a distinct neuronal population. In an effort to identify the regulatory elements that govern cerebellar granule cell-specific gene expression, the proximal 5' flanking regions for the human, rat, and mouse alpha6 genes were cloned and sequenced, and a major transcriptional initiation site was identified in the rodent genes. Functional analysis of rat alpha6 gene-reporter constructs in primary neuronal cultures reveals that a 155-bp TATA-less promoter region ( 130 to +25 bp) constitutes a minimal promoter that can drive cerebellar granule cell-specific expression. Internal deletion and decoy competition studies demonstrate that the minimal promoter contains a 60-bp region (-130 to -70 bp) that is critical for enhanced promoter activity in cerebellar granule cells. Activity of the compromised promoter containing the deletion cannot be rescued by placing the 60-bp region downstream of the reporter gene, demonstrating that it is not a classical enhancer but rather a positionally dependent regulator. An additional cerebellar-specific activating sequence is located between -324 and 130 bp, and a downstream negative regulatory region (+158 to +294) has been shown to be active in fibroblasts but inactive in cerebellar granule cells. Taken together, the results suggest a possible mechanism for the control of cerebellar granule cell-specific expression of the GABA(A) receptor alpha6 subunit gene. PMID- 10800929 TI - Opposite regulation of calbindin and calretinin expression by brain-derived neurotrophic factor in cortical neurons. AB - Regulation of calbindin and calretinin expression by brain-derived neurotrophic factor (BDNF) was examined in primary cultures of cortical neurons using immunocytochemistry and northern blot analysis. Here we report that regulation of calretinin expression by BDNF is in marked contrast to that of calbindin. Indeed, chronic exposure of cultured cortical neurons for 5 days to increasing concentrations of BDNF (0.1-10 ng/ml) resulted in a concentration-dependent decrease in the number of calretinin-positive neurons and a concentration dependent increase in the number of calbindin-immunoreactive neurons. Consistent with the immunocytochemical analysis, BDNF reduced calretinin mRNA levels and up regulated calbindin mRNA expression, providing evidence that modifications in gene expression accounted for the changes in the number of calretinin- and calbindin-containing neurons. Among other members of the neurotrophin family, neurotrophin-4 (NT-4), which also acts by activating tyrosine kinase TrkB receptors, exerted effects comparable to those of BDNF, whereas nerve growth factor (NGF) was ineffective. As for BDNF and NT-4, incubation of cortical neurons with neurotrophin-3 (NT-3) also led to a decrease in calretinin expression. However, in contrast to BDNF and NT-4, NT-3 did not affect calbindin expression. Double-labeling experiments evidenced that calretinin- and calbindin containing neurons belong to distinct neuronal subpopulations, suggesting that BDNF and NT-4 exert opposite effects according to the neurochemical phenotype of the target cell. PMID- 10800930 TI - Mutant presenilin 1 increases the levels of Alzheimer amyloid beta-peptide Abeta42 in late compartments of the constitutive secretory pathway. AB - Mutations in the presenilin 1 (PS1) gene are associated with autosomal dominant, early-onset, familial Alzheimer's disease and result in increased release of the hyperaggregatable 42-amino acid form of the amyloid beta-peptide (A(beta)42). To determine which subcellular compartments are potential source(s) of released Abeta42, we compared the levels and spatial segregation of intracellular A(beta)40 and A(beta)42 peptides between N2a neuroblastoma cells doubly transfected with the "Swedish" familial Alzheimer's disease-linked amyloid precursor protein variant and either wild-type PS1 (PS1(wt)) or familial Alzheimer's disease-linked delta9 mutant PS1 (PS1delta9). As expected, PS1delta9 expressing cells had dramatically higher levels of intracellular Abeta42 than did cells expressing PS1wt. However, the highest levels of A(beta)42 colocalized not with endoplasmic reticulum or Golgi markers but with rab8, a marker for trans Golgi network (TGN)-to-plasma membrane (PM) transport vesicles. We show that PS1 mutants are capable of causing accumulation of A(beta)42 in late compartments of the secretory pathway, generating there a readily releasable source of A(beta)42. Our findings indicate that PS1 "bioactivity" localizes to the vicinity of the TGN and/or PM and reconcile the apparent discrepancy between the preponderant concentration of PS1 protein in proximal compartments of the secretory pathway and the recent findings that PS1 "bioactivity" can control gamma-secretase-like processing of another transmembrane substrate, Notch, at or near the PM. PMID- 10800931 TI - L-arginine uptake and release by cultured avian retinal cells: differential cellular localization in relation to nitric oxide synthase. AB - The availability of L-arginine is of pivotal importance for the synthesis of nitric oxide, a signaling molecule in the CNS. Here we show the presence of a high-affinity L-arginine uptake system (Km of 4.4 +/- 0.5 microM and a Vmax of 26.0 +/- 0.9 fmol/well/min) in cultured chick retinal cells. Different compounds, such as N(G)-mono-methyl-L-arginine and L-lysine, were able to inhibit the uptake that was also inhibited 60-70% in the absence of sodium and/or calcium ions. No trans stimulation was observed when cells were preloaded with L-lysine. The data indicate that the L-arginine uptake in cultured retinal cells is partially mediated by the y+ system, but has a great contribution of the B(0,+) system. Autoradiographic studies revealed that the uptake is predominant in glial cells and can also be detected in neurons, whereas immunocytochemistry of nitric oxide synthase and L-citrulline showed that the enzyme is present in neurons and photoreceptors, but not in glial cells. L-[3H]Arginine is released from purified glial cultures incubated with high concentrations of potassium in the extracellular medium. Moreover, the amino acid released from preloaded glial cells was taken up by purified neuronal cultures. These results indicate that L arginine released from glial cells is taken up by neurons and used as substrate for the synthesis of nitric oxide. PMID- 10800932 TI - Development and characterisation of a glutamate-sensitive motor neurone cell line. AB - Modification of the growth conditions of NSC-34 mouse neuroblastoma x motor neurone cells by serum depletion promotes the expression of functional glutamate receptors as the cells mature into a form that bears the phenotypic characterisation of motor neurones. Immunocytochemical studies demonstrated the presence of the glutamate receptor proteins NMDAR1, NMDAR2A/B, GluR1, GluR2, GluR2/3, GluR4, GluR6/7, and KA2. Toxicity assays using cell counting techniques demonstrated a mild but significant cell death (approximately 30%, p < 0.01) following a 24-h exposure to 1 mM glutamate that could be prevented by the presence of the glutamate receptor antagonists (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine maleate (10 microM) and 2,3-dioxo-6-nitro 1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulphonamide (1 microM). As an indication of glutamate receptor functional activity a novel approach was used to detect the production of free radicals following stimulation with glutamate receptor agonists. The release of superoxide free radicals was detected using a micro electrochemical sensor following addition of glutamate receptor agonists to the cell bathing solution. Alterations in intracellular calcium concentrations were examined using fura-2 imaging. Exposure of the differentiated NSC-34 cells to glutamate leads to an increase in intracellular calcium concentrations that is prevented by the presence of glutamate receptor antagonists. The motor neurone origin of these cells makes them particularly useful for investigating the potential role of glutamatergic toxicity in motor neurone degeneration. PMID- 10800933 TI - High extracellular potassium modulates nitric oxide synthase expression in human astrocytes. AB - Inducible nitric oxide synthase (iNOS) is a molecule of great interest, given the numerous biological activities of nitric oxide and the documented expression of iNOS in several CNS pathologies. There also appears to be species-dependent regulation of iNOS expression as well as CNS-specific regulation. In this study, we have examined cultures of cytokine-activated primary human astrocytes as a model system with which to study the mechanisms of iNOS regulation in human CNS. As one of the major functions of astrocytes is spatial buffering of K+ ion, we examined the effect of high extracellular KCI on astrocyte iNOS expression. The results demonstrate that KCI at 25-75 mM potently inhibits astrocyte nitrite production stimulated by interleukin-1 (IL-1)/interferon-gamma (IFNgamma). In addition, several potassium channel inhibitors such as CsCl, tetraethylammonium, and 4-aminopyridine as well as nigericin inhibited astrocyte iNOS expression induced by IL-1/IFNgamma. These results demonstrate a novel role for astrocyte potassium channel activity in modulation of astrocyte function. They further suggest neural-specific mechanisms for glial iNOS regulation. PMID- 10800934 TI - Increase of brain-derived neurotrophic factor gene expression in NG108-15 cells by the nuclear isoforms of Ca2+/ calmodulin-dependent protein kinase II. AB - We have reported that the delta3 isoform of Ca2+/ calmodulin-dependent protein kinase II (CaM kinase II) is abundant in the nucleus in cerebellar granule cells. To examine the possibility that the nuclear isoforms of CaM kinase II are involved in the expression of brain-derived neurotrophic factor (BDNF), we transiently overexpressed the delta3 isoform in NG108-15 cells. The quantitative RT-PCR analysis revealed that rat cerebellum and NG108-15 cells expressed the exon IV-containing mRNA of BDNF (exon IV-BDNF mRNA) more than the exon III-BDNF mRNA. Treatment of NG108-15 cells with Bay K 8644 increased both exon III- and exon IV-BDNF mRNAs, and overexpression of the 83 isoform potentiated the expression of the exon IV-BDNF mRNA. The potentiation was not observed in the cells that were overexpressed with either the 61 isoform, a nonnuclear isoform, or the inactive mutant of the delta3 isoform. We constructed the luciferase reporter gene following the promoter upstream of exon IV and confirmed that overexpression of the delta3 isoform increased luciferase gene expression. Double immunostaining of NG108-15 cells with the antibodies to CaM kinase II and BDNF clearly showed that BDNF was highly expressed in the cells that were overexpressed with the delta3 isoform or the alphaB isoform, another nuclear isoform of CaM kinase II. These results suggest that the nuclear isoforms of CaM kinase II are involved in the expression of BDNF. PMID- 10800935 TI - Neurotrophin-3 controls proliferation of granular precursors as well as survival of mature granule neurons in the developing rat cerebellum. AB - Levels of neurotrophin-3 markedly decrease in the rat cerebellum after the first 10 days of life, suggesting an importance during early development. To further examine the effect of neurotrophin-3 on the developing cerebellum, we injected a monoclonal antibody against neurotrophin-3 into the lateral ventricle of 7.5-day old rats. The resultant depletion of neurotrophin-3 caused a significant decrease in cerebellar wet weights noted at 7 and 23 days thereafter. Other changes noted 48 h after injection of monoclonal antibodies against neurotrophin-3 included reduced incorporation of bromode-oxyuridine into granule neurons in the external germinal layer, an elevated density of atrophic neurons that had just migrated under the Purkinje cell layer, and an increased number of apoptotic neurons in the internal granule cell layer. These changes were limited to the central lobe. The concentration of neurotrophin-3 protein in the posterior region, including the central lobe, was about four- and threefold higher than that in the anterior region of the cerebellum of 9.5- and 30-day-old rats, respectively. Immunocytochemical examination showed higher amounts of neurotrophin-3 protein in the central lobe than in the anterior lobe. Our results provide evidence that neurotrophin-3 regulates the proliferation of granule precursors and supports the survival of mature granule neurons in restricted lobules, suggesting an involvement in limited regions at a specific stage in development of the rat cerebellum. PMID- 10800936 TI - Glutamate-stimulated activation of DNA synthesis via mitogen-activated protein kinase in primary astrocytes: involvement of protein kinase C and related adhesion focal tyrosine kinase. AB - Glutamate is the major excitatory neurotransmitter in the CNS. Although its role in neurons has been studied extensively, little is known about its function in astrocytes. We studied the effects of glutamate on signaling pathways in primary astrocytes. We found that the tyrosine kinase related adhesion focal tyrosine kinase (RAFTK) is tyrosine phosphorylated in response to glutamate in a time- and dose-dependent manner. This phosphorylation was pertussis toxin (PTX) sensitive and could be attenuated by the depletion of Ca2+ from intracellular stores. RAFTK tyrosine phosphorylation was mediated primarily by class I/II metabotropic glutamate receptors and depends on protein kinase C (PKC) activation. Glutamate treatment of primary astrocytes also results in a significant increase in the activity of the mitogen-activated protein kinases [extracellular signal-related kinases 1/2 (ERK1/2)]. Like RAFTK phosphorylation, ERK1/2 activation is PTX sensitive and can be attenuated by the depletion of intracellular Ca2+ and by PKC inhibition, suggesting that RAFTK might mediate the glutamate-dependent activation of ERK1/2. Furthermore, we demonstrated that glutamate stimulation of primary astrocytes leads to a significant increase in DNA synthesis. Glutamate stimulated DNA synthesis is PTX sensitive and can be inhibited by the MAP kinase kinase inhibitor PD98059, suggesting that in primary astrocytes, glutamate might signal via RAFTK and MAP kinase to promote DNA synthesis and cell proliferation. PMID- 10800937 TI - Tetanus toxin fragment C binds to a protein present in neuronal cell lines and motoneurons. AB - Tetanus Toxin Fragment C Binds to a Protein Present in Neuronal Cell Lines and Motoneurons Tetanus neurotoxin is one of the most powerful protein toxins known, acting in vivo at femtomolar doses. Two main factors determine its high potency: a protease activity restricted to a single intracellular substrate and its absolute neurospecificity. Whereas the enzymatic properties of tetanus toxin have been thoroughly defined, the nature of its neuronal receptor(s) and their involvement in the intracellular trafficking of tetanus toxin are poorly understood. Using binding and crosslinking experiments, we report here on the characterisation of an N-glycosylated 15-kDa interacting protein, which behaves as an integral membrane protein. This putative receptor specifically interacts with the binding domain (fragment C) of tetanus toxin and not with several related botulinum neurotoxins in spinal cord motoneurons and neuronal-like cell lines. Sialic acid-specific lectins antagonise the binding of tetanus toxin to the cell surface and to the 15-kDa protein, supporting the central role of sialic acid residues in the recognition process. Altogether, these results indicate the existence of a neuronal protein receptor for tetanus toxin whose identification is likely to constitute a key step in the analysis of the molecular machinery involved in the toxin internalisation and retrograde transport. PMID- 10800938 TI - Stretch-induced injury alters mitochondrial membrane potential and cellular ATP in cultured astrocytes and neurons. AB - Energy deficit after traumatic brain injury (TBI) may alter ionic homeostasis, neurotransmission, biosynthesis, and cellular transport. Using an in vitro model for TBI, we tested the hypothesis that stretch-induced injury alters mitochondrial membrane potential (delta(psi)m) and ATP in astrocytes and neurons. Astrocytes, pure neuronal cultures, and mixed neuronal plus glial cultures grown on Silastic membranes were subjected to mild, moderate, and severe stretch. After injury, delta(psi)m was measured using rhodamine-123, and ATP was quantified with a luciferin-luciferase assay. In astrocytes, delta(psi)m dropped significantly, and ATP content declined 43-52% 15 min after mild or moderate stretch but recovered by 24 h. In pure neurons, delta(psi)m declined at 15 min only in the severely stretched group. At 48 h postinjury, delta(psi)m remained decreased in severely stretched neurons and dropped in moderately stretched neurons. Intracellular ATP content did not change in any group of injured pure neurons. We also found that astrocytes and neurons release ATP extracellularly following injury. In contrast to pure neurons, delta(psi)m in neurons of mixed neuronal plus glial cultures declined 15 min after mild, moderate, or severe stretch and recovered by 24-48 h. ATP content in mixed cultures declined 22-28% after mild to severe stretch with recovery by 24 h. Our findings demonstrate that injury causes mitochondrial dysfunction in astrocytes and suggest that astrocyte injury alters mitochondrial function in local neurons. PMID- 10800939 TI - Regulation of tryptophan hydroxylase activity in Xenopus laevis photoreceptor cells by cyclic AMP. AB - The aim of this study was to investigate the role of cyclic AMP in the regulation of tryptophan hydroxylase activity localized in retinal photoreceptor cells of Xenopus laevis, where the enzyme plays a key role in circadian melatonin biosynthesis. In photoreceptor-enriched retinas that lack serotonergic neurons, tryptophan hydroxylase activity is markedly stimulated by treatments that increase intracellular levels of cyclic AMP or activate cyclic AMP-dependent protein kinase, including forskolin, phosphodiesterase inhibitors, and cyclic AMP analogues. In contrast, cyclic AMP has no effect on tryptophan hydroxylase mRNA abundance. Experiments using cycloheximide and actinomycin D demonstrate that cyclic AMP exerts its regulatory effect via posttranslational mechanisms mediated by cyclic AMP-dependent protein kinase. The effect of cyclic AMP is independent of the phase of the photoperiod, suggesting that the nucleotide is not a mediator of the circadian rhythm of tryptophan hydroxylase. Cyclic AMP accumulation is higher in darkness than in light, as is tryptophan hydroxylase activity. Furthermore, the stimulatory effect of forskolin and that of darkness are inhibited by H89, an inhibitor of cyclic AMP-dependent protein kinase. In conclusion, cyclic AMP may mediate the acute effects of light and darkness on tryptophan hydroxylase activity of retinal photoreceptor cells. PMID- 10800940 TI - Protective effect of the energy precursor creatine against toxicity of glutamate and beta-amyloid in rat hippocampal neurons. AB - The loss of ATP, which is needed for ionic homeostasis, is an early event in the neurotoxicity of glutamate and beta-amyloid (A(beta)). We hypothesize that cells supplemented with the precursor creatine make more phosphocreatine (PCr) and create larger energy reserves with consequent neuroprotection against stressors. In serum-free cultures, glutamate at 0.5-1 mM was toxic to embryonic hippocampal neurons. Creatine at >0.1 mM greatly reduced glutamate toxicity. Creatine (1 mM) could be added as late as 2 h after glutamate to achieve protection at 24 h. In association with neurotoxic protection by creatine during the first 4 h, PCr levels remained constant, and PCr/ATP ratios increased. Morphologically, creatine protected against glutamate-induced dendritic pruning. Toxicity in embryonic neurons exposed to A(beta) (25-35) for 48 h was partially prevented by creatine as well. During the first 6 h of treatment with A(beta) plus creatine, the molar ratio of PCr/ATP in neurons increased from 15 to 60. Neurons from adult rats were also partially protected from a 24-h exposure to A(beta) (25-35) by creatine, but protection was reduced in neurons from old animals. These results suggest that fortified energy reserves are able to protect neurons against important cytotoxic agents. The oral availability of creatine may benefit patients with neurodegenerative diseases. PMID- 10800941 TI - A late phase of exocytosis from synaptosomes induced by elevated [Ca2+]i is not blocked by Clostridial neurotoxins. AB - Treatment of rat cerebrocortical synaptosomes with botulinum toxin types E and C1 or tetanus toxin removed the majority of intact SNAP-25, syntaxin 1A/1B, and synaptobrevin and diminished Ca(2+)-dependent K+ depolarization-induced noradrenaline secretion. With botulinum toxin type E, <10% of intact SNAP-25 remained and K(+)-evoked release of glutamate and GABA was inhibited. The large component of noradrenaline release evoked within 120 s by inclusion of the Ca2+ ionophore A23187 with the K+ stimulus was also attenuated by these toxins; additionally, botulinium neurotoxin type E blocked the first 60 s of ionophore induced GABA and glutamate exocytosis. However, exposure to A23187 for longer periods induced a phase of secretion nonsusceptible to any of these toxins (>120 s for noradrenaline; >60 s for glutamate or GABA). Most of this late phase of release represented exocytosis because of its Ca2+ dependency, ATP requirement, and sensitivity to a phosphatidylinositol 4-kinase inhibitor. Based on these collective findings, we suggest that the ionophore-induced elevation of [Ca2+]i culminates in the disassembly of complexes containing nonproteolyzed SNAP receptors protected from the toxins that can then contribute to neuroexocytosis. PMID- 10800942 TI - Synergistic depletion of astrocytic glutathione by glucose deprivation and peroxynitrite: correlation with mitochondrial dysfunction and subsequent cell death. AB - Previously we reported that immunostimulated astrocytes were highly vulnerable to glucose deprivation. The augmented death was mimicked by the peroxynitrite (ONOO )-producing reagent 3-morpholinosydnonimine (SIN-1). Here we show that glucose deprivation and ONOO- synergistically deplete intracellular reduced glutathione (GSH) and augment the death of astrocytes via formation of cyclosporin A sensitive mitochondrial permeability transition (MPT) pore. Astrocytic GSH levels were only slightly decreased by glucose deprivation or SIN-1 (200 microM) alone. In contrast, a rapid and large depletion of GSH was observed in glucose-deprived/ SIN-1-treated astrocytes. The depletion of GSH occurred before a significant release of lactate dehydrogenase (a marker of cell death). Superoxide dismutase and ONOO-scavengers completely blocked the augmented death, indicating that the reaction of nitric oxide with superoxide to form ONOO was implicated. Furthermore, nitrotyrosine immunoreactivity (a marker of ONOO-) was markedly enhanced in glucose-deprived/SIN-1 -treated astrocytes. Mitochondrial transmembrane potential (MTP) was synergistically decreased in glucose deprived/SIN-1-treated astrocytes. The glutathione synthase inhibitor L buthionine-(S,R)-sulfoximine markedly decreased the MTP and increased lactate dehydrogenase (LDH) releases in SIN-1-treated astrocytes. Cyclosporin A, an MPT pore blocker, completely prevented the MTP depolarization as well as the enhanced LDH releases in glucose-deprived/SIN-1-treated astrocytes. PMID- 10800943 TI - Characteristics of the calcium-triggered mitochondrial permeability transition in nonsynaptic brain mitochondria: effect of cyclosporin A and ubiquinone O. AB - The objective of the present study was to assess the capacity of nonsynaptic brain mitochondria to accumulate Ca2+ when subjected to repeated Ca2+ loads, and to explore under what conditions a mitochondrial permeability transition (MPT) pore is assembled. The effects of cyclosporin A (CsA) on Ca2+ accumulation and MPT pore assembly were compared with those obtained with ubiquinone 0 (Ubo), a quinone that is a stronger MPT blocker than CsA, when tested on muscle and liver mitochondria. When suspended in a solution containing phosphate (2 mM) and Mg2+ (1 mM), but no ATP or ADP, the brain mitochondria had a limited capacity to accumulate Ca2+ (210 nmol/mg of mitochondrial protein). Furthermore, when repeated Ca2+ pulses (40 nmol/mg of protein each) saturated the uptake system, the mitochondria failed to release the Ca2+ accumulated. However, in each instance, the first Ca2+ pulse was accompanied by a moderate release of Ca2+, a release that was not observed during the subsequent pulses. The initial release was accompanied by a relatively marked depolarization, and by swelling, as assessed by light-scattering measurements. However, as the swelling was <50% of that observed following addition of alamethicin, it is concluded that the first Ca2+ pulse gives rise to an MPT in a subfraction of the mitochondrial population. CsA, an avid blocker of the MPT pore, only marginally increased the Ca(2+) sequestrating capacity of the mitochondria. However, CsA eliminated the Ca2+ release accompanying the first Ca2+ pulse. The effects of CsA were shared by Ubo, but when the concentration of Ubo exceeded 20 microM, it proved toxic. The results thus suggest that brain mitochondria are different from those derived from a variety of other sources. The major difference is that a fraction of the brain mitochondria, studied presently, depolarized and showed signs of an MPT. This fraction, but not the remaining ones, contributed to the chemically and electron microscopically verified mitochondrial swelling. PMID- 10800944 TI - A role for the small molecular weight GTPases, Rho and Cdc42, in muscarinic receptor signaling to focal adhesion kinase. AB - An enhanced tyrosine phosphorylation of focal adhesion kinase (FAK) is elicited during neuronal growth cone remodeling and requires the maintenance of agonist sensitive pools of phosphatidylinositol 4,5-bisphosphate (PIP2). Rho family GTPases are putative regulators of both PIP2 synthesis and growth cone remodeling, including neurite outgrowth elicited by muscarinic cholinergic receptor (mAChR) stimulation. In this study, we investigated the interrelationships among Rho family GTPases, PIP2 synthesis, and mAChR signaling to FAK in SH-SY5Y neuroblastoma cells. Preincubation with Clostridium difficile toxin B (Tox B), an inhibitor of Rho, Rac, and Cdc42, attenuated mAChR-stimulated FAK and paxillin tyrosine phosphorylation and lysophosphatidic acid (LPA)-induced FAK phosphorylation to a similar extent (75% decreases at 200 pg/ml Tox B) but did not affect mitogen-activated protein kinase activation elicited by either phorbol ester or an mAChR agonist. In contrast, preincubation with selective inhibitors of either Rho (C3 exoenzyme) or Rho kinase (HA-1 077) resulted in 80 90% reductions in LPA-induced FAK phosphorylation but only 40-50% decreases in mAChR-stimulated phosphorylation. Moreover, mAChR-mediated FAK phosphorylation was significantly attenuated in cells scrape-loaded with dominant-negative N17Cdc42 but not N17Rac1. Tox B had little or no effect on agonist-sensitive pools of PIP2 but inhibited mAChR-driven actin cytoskeletal remodeling. The results suggest that the Rho family GTPases, Rho and Cdc42, link mAChR stimulation to increases in FAK phosphorylation independently of effects on PIP2 synthesis. PMID- 10800945 TI - Differential regulation of leukemia inhibitory factor-stimulated neuronal gene expression by protein phosphatases SHP-1 and SHP-2 through mitogen-activated protein kinase-dependent and -independent pathways. AB - The neurally active cytokine leukemia inhibitory factor (LIF) signals through a bipartite receptor complex composed of LIF receptor alpha (LIFR) and gp130. gp130 and LIFR contain consensus binding motifs for the protein tyrosine phosphatase SHP-2 surrounding tyrosines 118 and 115 (Y118 and Y115) of their cytoplasmic domains, respectively. These sites are necessary for maximal activation of mitogen-activated protein kinase (MAPK). Coexpression of catalytically inactive, but not wild-type, SHP-2 reduced LIFR- and gp130-mediated activation of MAPK up to 75%. Conversely, coexpression of the wild-type, but not catalytically inactive, SHP-1, a related phosphatase, reduced activity up to 80%, demonstrating that SHP-2 and SHP-1 have opposing effects on the MAPK pathway. Mutation of Y115 of the cytoplasmic domain of LIFR eliminates receptor-mediated tyrosine phosphorylation of SHP-2. In contrast, SHP-1 association with gp130 and LIFR is constitutive and independent of Y118 and Y115, respectively. SHP-1 has a positive regulatory role on LIF-stimulated vasoactive intestinal peptide (VIP) reporter gene expression in neuronal cells, whereas the effect of SHP-2 is negative. Furthermore, LIF-stimulated MAPK activation negatively regulates this VIP reporter gene induction. SHP-2 also negatively regulates LIF-dependent expression of choline acetyltransferase, but this regulation could be dissociated from its effects on MAPK activation. These data indicate that SHP-1 and SHP-2 are important regulators of LIF-dependent neuronal gene expression via both MAPK dependent and -independent pathways. PMID- 10800946 TI - Inflammatory cytokines enhance muscarinic-mediated arachidonic acid release through p38 mitogen-activated protein kinase in A2058 cells. AB - The human melanoma cell line A2058 expresses the Gq-coupled M5 subtype of muscarinic receptor. Stimulation with the cholinergic agonist, carbachol, induces a dose-dependent increase in arachidonic acid release. The carbachol-induced arachidonate release is potentiated two- to threefold by pretreatment of A2058 cells with either of the inflammatory cytokines, tumor necrosis factor-alpha or interleukin-1beta . Cytokine-induced enhancement of muscarinic-mediated arachidonic acid release peaks near 1 h. Western analysis suggests that both cytokines are capable of activating the nuclear factor-kappaB (NF-kappaB) and p38 mitogen-activated protein kinase (MAPK) pathways. Anisomycin (1 microM) treatment mimics the cytokine-induced enhancement of arachidonic acid production and activates the p38 MAPK pathway, but does not activate the NF-kappaB pathway. Furthermore, pre-treatment of A2058 cells with the putative p38 MAPK inhibitor, SB202190, ablates the cytokine-dependent augmentation without interfering with the muscarinic-mediated arachidonic acid release in untreated cells. Moreover, cytokine treatment does not affect other M5-coupled pathways (e.g., phospholipase C activity or intracellular Ca2+ mobilization), suggesting that p38 MAPK activation principally modulates muscarinic-mediated phospholipase A2 activity. Finally, in primary cultures of cells taken from rat cerebellum, key aspects of this finding are repeated in cultures enriched for glia, but not in cultures enriched for granule neurons. PMID- 10800947 TI - Implication of glutamate in the expression of inducible nitric oxide synthase after oxygen and glucose deprivation in rat forebrain slices. AB - Nitric oxide synthesis by inducible nitric oxide synthase (iNOS) has been postulated to contribute to ischemia-reperfusion neurotoxicity. The expression of this enzyme has been demonstrated in cells present in the postischemic brain. The mechanisms of iNOS expression after cerebral ischemia are a subject of current research. We therefore decided to investigate whether glutamate, which is released in ischemia and is implicated in neurotoxicity, might be involved in the mechanisms by which oxygen and glucose deprivation (OGD) leads to the expression of iNOS in rat forebrain slices. In this model, we have shown previously that 20 min of OGD causes the expression of iNOS. We have now found that the NMDA receptor antagonist MK-801 blocks the expression of iNOS, suggesting that the activation of the NMDA subtype of glutamate receptor is implicated in the mechanisms that lead to the expression of this isoform. Moreover, we have found that glutamate alone could trigger the induction process, as shown by the appearance of a Ca(2+)-independent NOS activity and by the detection of iNOS mRNA and protein in slices exposed to glutamate. Glutamate-dependent iNOS expression was concentration-dependent and was blocked by EGTA and by the inhibitors of nuclear factor kappaB (NF-kappaB) activation pyrrolidine dithiocarbamate and MG132. In addition, glutamate induced NF-kappaB translocation to the nucleus, an effect that was inhibited by MG132. Taken together, our data suggest that activation of NMDA receptors by glutamate released in ischemia is involved in the expression of iNOS in rat forebrain slices via a Ca(2+)-dependent activation of the transcription factor NF-kappaB. To our knowledge, this is the first report showing an implication of excitatory amino acids in the expression of iNOS caused by ischemia. PMID- 10800948 TI - Effect of intracerebral administration of NMDA and AMPA on dopamine and glutamate release in the ventral pallidum and on motor behavior. AB - The present study investigates the modulation of the ventral tegmental area (VTA) ventral pallidum (VP) dopaminergic system by glutamate agonists in rats. The glutamate receptor agonists N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid (AMPA) were infused via reversed microdialysis into the VTA, and dopamine (DA), glutamate, and aspartate levels in the VTA and ipsilateral VP were monitored together with motor behavior screened in an open field. NMDA (750 microM) infusion, as well as AMPA (50 microM) infusion, induced an increase of DA and glutamate levels in the VTA, followed by an increase of DA levels in the ipsilateral VP and by enhanced locomotor activity. The increase of DA in the VP was similar after administration of these two glutamate agonists, although motor activity was more pronounced and showed an earlier onset after NMDA infusion. Glutamate levels in the VP were not increased by the stimulation of DA release. It is concluded that DA is released from mesencephalic DA neurons projecting to the VP and that these neurons are controlled by glutamatergic systems, via NMDA and AMPA receptors. Thus, DA in the VP has to be considered as a substantial modulator. Dysregulation of the mesopallidal DA neurons, as well as their glutamatergic control, may play an additional or distinct role in disorders like schizophrenia and drug addiction. PMID- 10800949 TI - Nerve growth factor-induced phosphorylation of SNAP-25 in PC12 cells: a possible involvement in the regulation of SNAP-25 localization. AB - Synaptosomal-associated protein of 25 kDa (SNAP-25), a t-SNARE protein essential for neurotransmitter release, is phosphorylated at Ser187 following activation of cellular protein kinase C by treatment with phorbol 12-myristate 13-acetate. However, it remains unclear whether neuronal activity or an endogenous ligand induces the phosphorylation of SNAP-25. Here we studied the phosphorylation of SNAP-25 in PC12 cells using a specific antibody for SNAP-25 phosphorylated at Ser187. A small fraction of SNAP-25 was phosphorylated when cells were grown in the absence of nerve growth factor (NGF). A brief treatment with NGF that was enough to activate the mitogen-activated protein kinase signal transduction pathway did not increase the phosphorylation of SNAP-25; however, phosphorylation was up-regulated after a prolonged incubation with NGF. Up-regulation was transitory, and maximum phosphorylation (a fourfold increase over basal phosphorylation) was achieved between 36 and 48 h after the addition of NGF. Immunofluorescent microscopy showed that SNAP-25 was localized primarily in the plasma membrane, although a significant population was also present in the cytoplasm. Quantitative microfluorometry revealed that prolonged treatment with NGF resulted in a preferential localization of SNAP-25 in the plasma membrane. A mutational study using a fusion protein with green fluorescent protein as a tag indicated that the point mutation of Ser187 to Ala abolished the NGF-dependent relocalization. A population of SNAP-25 in the plasma membrane was not increased by a point mutation at Ser187 to Glu; however, it was increased by prolonged treatment with NGF, indicating that the SNAP-25 phosphorylation is essential, but not sufficient, for the NGF-induced relocation to the plasma membrane. Our results suggest a close temporal relationship between the up-regulation of SNAP 25 phosphorylation and its relocation, and NGF-induced differentiation of PC12 cells. PMID- 10800950 TI - p-ethynylphenylalanine: a potent inhibitor of tryptophan hydroxylase. AB - Tryptophan hydroxylase (TPH) is the initial and rate-limiting enzyme in serotonin biosynthesis. The enzyme activity is dependent on molecular oxygen, a tetrahydropterin cosubstrate, and ferrous iron. The present study demonstrates that TPH is inhibited by a novel compound, p-ethynylphenylalanine (pEPA), produced by the Heck reaction of trimethylsilylacetylene with N tertbutyloxycarbonyl-4-iodo-L-phenylalanine methyl ester. pEPA is a more potent and specific inhibitor of TPH than p-chlorophenylalanine (pCPA). In the present study, pEPA was demonstrated to inhibit competitively and reversibly TPH in vitro (Ki = 32.6 +/- 6.2 microM vs. tryptophan). pEPA displayed little inhibitory activity toward tyrosine hydroxylase (EC 1.14.16.2), the initial and rate limiting enzyme for catecholamine biosynthesis, and no inhibition of phenylalanine hydroxylase or tyrosinase. In addition, pEPA was a poor ligand for the serotonin transporter and several serotonin receptors. Administration of pEPA (30 mg/kg) to rats produced a 95 +/- 5% decrease in TPH activity in brain homogenates and a concomitant decrease in serotonin and 5-hydroxyindole-3-acetic acid levels (85%) at 24 h after injection. In contrast, pCPA produced a similar effect (87 +/- 5% decrease in TPH activity) only at 10 times the concentration (300 mg/kg). These results suggest that pEPA is a selective, reversible, and potent inhibitor of TPH both in vitro and in vivo. The potential for pEPA to inhibit selectively and reversibly the biosynthesis of serotonin may contribute to the characterization of the role of serotonin in behavioral and physiological activities. PMID- 10800951 TI - The activity-regulated cytoskeletal-associated protein arc is expressed in different striosome-matrix patterns following exposure to amphetamine and cocaine. AB - The activity-regulated, cytoskeletal-associated gene, arc, is a brain-enriched immediate-early gene whose expression is rapidly induced in the striatum by dopamine receptor agonists. This rapid induction of arc in the striatum is similar to that of other early response genes such as c-fos, junB, deltafosB, fra, and NGFI-A, which code for transcription factors. Unlike these proteins, however, Arc is a cytoskeletal protein expressed not only in the nucleus of neurons but also in their dendrites. We investigated the patterns of Arc expression evoked in the rat striatum by acute exposures to two psychomotor stimulants, cocaine and amphetamine. Cocaine induced arc in striatal neurons that were broadly distributed within both striosome and matrix compartments of the caudoputamen. Amphetamine also evoked Arc expression in striatal projection neurons, but these were heavily concentrated in the striosomal compartment and only sparsely in the matrix compartment in the rostral striatum. The contrasting patterns of Arc expression evoked by cocaine and amphetamine parallel those of c Fos, JunB, FRA, and NGFI-A expression induced by these two psychomotor stimulants. This difference in the action of cocaine and amphetamine at the level of protein expression may be linked to the different effects of these psychomotor stimulants on behavior. PMID- 10800952 TI - In vitro antioxidant neuroprotective activity of BN 80933, a dual inhibitor of neuronal nitric oxide synthase and lipid peroxidation. AB - BN 80933, a dual inhibitor of neuronal nitric oxide synthase and lipid peroxidation, prevents in vivo brain ischemic/reperfusion injury. In the present study, BN 80933 was shown to protect neurons from hypoxia-induced cell death in primary cultures of cortical neurons. BN 80933 prevented lactate dehydrogenase activity elevation induced by hypoxia, displaying an IC50 value of 0.15 +/- 0.05 microM. This effect was likely due to the antioxidant properties of BN 80933 because Trolox, but not NG-nitro-L-arginine, also elicited protection. The antioxidant property of BN 80933 was then further investigated on HT-22 cells subjected to buthionine sulfoximine- or glutamate-induced glutathione depletion. The relative order of potency of the various compounds to inhibit oxidative stress-induced neuronal death (BN 80933 > U104067 > butylated hydroxytoluene > 17beta-estradiol > Trolox > vitamin E) correlated with their ability to inhibit brain membrane lipid peroxidation (correlation coefficient = 0.939). BN 80933 afforded protection even when added 6 h after glutamate exposure. BN 80933 did not reverse intracellular glutathione depletion but prevented elevation of the level of beta-epiprostaglandin F2alpha (8-isoprostane), which appeared to be a delayed phenomenon. In conclusion, BN 80933 induces a potent cytoprotection that may be mediated by inhibition of delayed lipid peroxidation. PMID- 10800953 TI - Taurine-induced attenuation of MPP+ neurotoxicity in vitro: a possible role for the GABA(A) subclass of GABA receptors. AB - Taurine is a sulphur-containing beta-amino acid found in high (millimolar) concentrations in excitable tissues such as brain and heart. Its suggested roles include osmoregulator, thermoregulator, neuromodulator, and potential neurotransmitter. This amino acid has also been shown to be released in large concentrations during ischaemia and excitotoxin-induced neuronal damage. Here we report a protective effect of taurine against MPP(+)-induced neurotoxicity in coronal slices from rat brain. Significant protective effects were observed at taurine concentrations of 20 and 1 mM, suggesting a potential role for taurine in cases of neuronal insult. Studies with the synthetic taurine analogues taurine phosphonate, guanidinoethane sulphonate, and trimethyltaurine suggested the observed effect to be mediated via an extracellular mechanism. The use of GABA receptor ligands muscimol and bicuculline indicated the effect to be mediated through activation of GABA(A) receptors. PMID- 10800954 TI - Inhibition of noxious stimulus-induced spinal prostaglandin E2 release by flurbiprofen enantiomers: a microdialysis study. AB - Peripheral noxious stimuli have been shown to induce prostaglandin (PG) E2 release at the site of inflammation and in the spinal cord. The antiinflammatory and antinociceptive effects of cyclooxygenase-inhibiting drugs are thought to depend on the inhibition of PG synthesis. R-Flurbiprofen, however, does not inhibit cyclooxygenase activity in vitro but still produces antinociceptive effects. To find out whether R-flurbiprofen acts via inhibition of spinal PG release, concentrations of PGE2 and flurbiprofen in spinal cord tissue were assessed by microdialysis. The catheter was transversally implanted through the dorsal horns of the spinal cord at level L4. R- and S-flurbiprofen (9 and 27 mg kg(-1), respectively) were administered intravenously 10-15 min before subcutaneous injection of formalin into the dorsal surface of one hindpaw. Flurbiprofen was rapidly distributed into the spinal cord with maximal concentrations after 30-45 min. Baseline PGE2 dialysate concentrations were 100.6 +/- 6.4 pg ml(-1) (mean +/- SEM). After formalin injection they rose about threefold with a maximum of 299.4 +/- 68.4 pg ml(-1) at 7.5 min. After approximately 1 h PGE2 levels returned to baseline. Both flurbiprofen enantiomers completely prevented the formalin-induced increase of spinal PGE2 release and reduced PGE2 concentrations below basal levels. S- and R-flurbiprofen at 9 mg kg( 1) produced a minimum of 15.8 +/- 5.2 and 27.7 +/- 14.9 pg ml(-1), respectively, and 27 mg kg(-1) S- and R-flurbiprofen resulted in 11.7 +/- 1.7 and 9.3 +/- 4.7 pg ml(-1), respectively. PGE2 levels remained at the minimum up to the end of the observation period at 5 h. When 27 mg kg(-1) R-flurbiprofen was injected intravenously without subsequent formalin challenge, baseline immunoreactive PGE2 concentrations were not affected. S-Flurbiprofen (27 mg kg(-1)), however, led to a moderate reduction (approximately 40%). The data suggest that antinociception produced by R-flurbiprofen is mediated at least in part by inhibition of stimulated spinal PGE2 release and support the current view that increased spinal PGE2 release significantly contributes to nociceptive processing. PMID- 10800955 TI - Calpain inhibitors confer biochemical, but not electrophysiological, protection against anoxia in rat optic nerves. AB - Calpains are ubiquitous Ca(2+)-activated neutral proteases that have been implicated in ischemic and traumatic CNS injury. Ischemia and trauma of central white matter are dependent on Ca2+ accumulation, and calpain overactivation likely plays a significant role in the pathogenesis. Adult rat optic nerves, representative central white matter tracts, were studied in an in vitro anoxic model. Functional recovery following 60 min of anoxia and reoxygenation was measured electrophysiologically. Calpain activation was assessed using western blots with antibodies against calpain-cleaved spectrin breakdown products. Sixty minutes of in vitro anoxia increased the amount of spectrin breakdown approximately 20-fold over control, with a further increase after reoxygenation to >70 times control, almost as much as 2 h of continuous anoxia. Blocking voltage-gated Na+ channels with tetrodotoxin or removing bath Ca2+ was highly neuroprotective electrophysiologically and resulted in a marked reduction of spectrin degradation. The membrane-permeable calpain inhibitors MDL 28,170 and calpain inhibitor-I (10-100 microM) were effective at reducing spectrin breakdown in anoxic and reoxygenated optic nerves, but no electrophysiological improvement was observed. We conclude that calpain activation is an important step in anoxic white matter injury, but inhibition of this Ca(2+)-dependent process in isolation does not improve functional outcome, probably because other deleterious Ca(2+) activated pathways proceed unchecked. PMID- 10800956 TI - Nonlinear decrease over time in N-acetyl aspartate levels in the absence of neuronal loss and increases in glutamine and glucose in transgenic Huntington's disease mice. AB - Mice transgenic for exon I of mutant huntingtin, with 141 CAG repeats, exhibit a profound symptomatology characterized by weight loss, motor disorders, and early death. We performed longitudinal analysis of metabolite levels in these mice using NMR spectroscopy in vivo and in vitro. These mice exhibited a large (53%), nonlinear drop in in vivo N-acetyl aspartate (NAA) levels over time, commencing at approximately 6 weeks of age, coincident with onset of symptoms. These drops in NAA levels occurred in the absence of neuronal death as measured by postmortem Nissl staining and neuronal counting but in the presence of nuclear inclusion bodies. In addition to decreased NAA, these mice showed a large elevation of glucose in the brain (600%) consistent with a diabetic profile and elevations in blood glucose levels both before and after glucose loading. In vitro NMR analysis revealed significant increases in glutamine (100%), taurine (95%) cholines (200%), and scyllo-inositol (333%) and decreases in glutamate (24%) and succinate (47%). These results lead to two conclusions. NAA is reflective of the health of neurons and thus is a noninvasive marker, with a temporal progression similar to nuclear inclusion bodies and symptoms, of neuronal dysfunction in transgenic mice. Second, the presence of elevated glutamine is evidence of a profound metabolic defect. We present arguments that the elevated glutamine results from a decrease in neuronal-glial glutamate-glutamine cycling and a decrease in glutaminase activity. PMID- 10800957 TI - The human nucleus accumbens is highly susceptible to G protein down-regulation by methamphetamine and heroin. AB - Although the nucleus accumbens is assumed to be a critical brain "pleasure center," its function in humans is unknown. As animal data suggest that a unique feature of this small brain area is its high sensitivity to down-regulation of an inhibitory G protein by drugs of abuse, we compared G protein levels in postmortem nucleus accumbens with those in seven other brain regions of chronic users of cocaine, methamphetamine, and heroin, and of matched controls. Biochemical changes were restricted to the nucleus accumbens in which concentrations of G(alpha)1 and/or G(alpha)2 were reduced by 32-49% in the methamphetamine and heroin users. This selective responsiveness to these abused drugs implies a special role for the human nucleus accumbens in mechanisms of drug reinforcement and suggests that some features of the drug-dependent state (e.g., tolerance) might be related to inhibition of G(alpha)1-linked receptor activity. PMID- 10800958 TI - Commercial recombinant human beta-nerve growth factor and adult rat dorsal root ganglia contain an identical molecular species of nerve growth factor prohormone. AB - Examination of commercial recombinant human beta-nerve growth factor (rh-beta NGF) preparations with polyclonal antibodies specific to 13-kDa NGF and pro-NGF specific domains revealed the presence of high-molecular-mass immunoreactive proteins, including a 60-kDa NGF prohormone. On incubation with a mixture of N- and O-specific glycosidases, the 60-kDa NGF pro-hormone generated a 32-kDa protein corresponding to the molecular size of NGF precursor predicted by the cloned human NGF cDNA. Highly sensitive chemiluminescence immunoblot analysis of adult rat dorsal root ganglia, spinal cord, and colon tissues with NGF- and pro NGF domain-specific antibodies also revealed the presence of high-molecular-mass proteins, including the 60-kDa NGF prohormone. Based on the presence of the 60 kDa NGF prohormone in dorsal root ganglia and its efferent tissues, we suggest that proteolytically unprocessed, glycosylated NGF prohormone may mediate interactions between neurons and the tissues they innervate. PMID- 10800959 TI - A functional analysis of EP4 receptor-expressing neurons in mediating the action of prostaglandin E2 within specific nuclei of the brain in response to circulating interleukin-1beta. AB - Proinflammatory cytokines released by cells of myeloid lineage have the ability to stimulate different populations of neurons through intermediate molecules released by cells of the blood-brain barrier. The aim of the present study was to verify the hypothesis that prostaglandins (PGs) play a site-specific role in activating selective groups of neurons via a privileged interaction between PG of the E2 type and its EP4 receptor. In a first set of experiments, animals were treated with the inhibitor of PG synthesis ketorolac to determine the endogenous contribution of PG in mediating the neuronal activation and EP4 expression in response to circulating interleukin-1beta(IL-1beta). The subsequent experiment consisted of evaluating the role of PGE2 in activating EP4-expressing neurons in the rat brain. Ketorolac completely abolished the endogenous release of PGE2 in the liver and prevented the induction of immediate-early genes and up-regulation of EP4 mRNA in specific groups of neurons, such as the parvocellular paraventricular nucleus and the A1 catecholaminergic population of cells. This effect was, however, not generalized throughout the brain as PGE2 inhibition failed to abolish IL-1beta-induced c-fos transcription in the nucleus of the solitary tract, parabrachial nucleus, bed nucleus of the stria terminalis, and the circumventricular organs. Of interest are the data that central PGE2 injection activated EP4 gene transcription in neurons that no longer responded to the intravenous IL-1beta bolus when the animals were pretreated with ketorolac. Site-specific interaction between the ligand and its receptor was further supported by the induction of c-fos-immunoreactive nuclei within EP4-expressing neurons in response to intracerebroventricular PGE2 infusion. Both intracerebroventricular PGE2 and intravenous IL-1beta injection provoked a sharp and rapid increase in plasma corticosterone levels, an effect that was completely prevented in inhibiting PG production in IL-1beta-challenged rats. These data provide the evidence that EP4 is expressed in numerous nuclei involved in autonomic and neuroendocrine control, although a privileged interaction seems to take place in specific nuclei and areas, including the endocrine hypothalamus and the A1 cell group of the ventrolateral medulla. It is quite possible that EP4 acts as the functional receptor for PGE2 to activate the neuronal circuit involved in the activation of the glucocorticoid axis, as an essential neuroendocrine response for the appropriate control of systemic inflammation. PMID- 10800960 TI - Hydrophobic protein that copurifies with human brain acetylcholinesterase: amino acid sequence, genomic organization, and chromosomal localization. AB - The mechanism of attachment of acetylcholinesterase (AChE) to neuronal membranes in interneuronal synapses is poorly understood. We have isolated, sequenced, and cloned a hydrophobic protein that copurifies with AChE from human caudate nucleus and that we propose forms a part of a complex of membrane proteins attached to this enzyme. It is a short protein of 136 amino acids and has a molecular mass of 18 kDa. The sequence contains stretches of both hydrophobic and hydrophilic amino acids and two cysteine residues. Analysis of the genomic sequence reveals that the coding region is divided among five short exons. Fluorescence in situ hybridization localizes the gene to chromosome 6p21.32-p21.2. Northern blot analysis shows that this gene is widely expressed in the brain with an expression pattern that parallels that of AChE. PMID- 10800961 TI - Mice homozygous for the L250T mutation in the alpha7 nicotinic acetylcholine receptor show increased neuronal apoptosis and die within 1 day of birth. AB - The alpha7 nicotinic acetylcholine receptor (nAChR) has been implicated in modulating neurotransmitter release and may play a role in the regulation of neuronal growth and differentiation. A threonine for leucine 247 substitution in the channel domain of the chick alpha7 nAChR increases agonist affinity and decreases the rate of desensitization, creating a "gain of function" model for this receptor. We have generated mice that express the analogous mutation (L250T) in the alpha7 nAChR using the techniques of homologous recombination and here report their characteristics. Mice heterozygous (+/T) for the L250T mutation are viable, fertile, and anatomically normal compared with wild-type littermates. In contrast, homozygous (T/T) L250T mice die within 2-24 h of birth. Brains of T/T mouse pups exhibit a marked reduction in alpha7 nAChR protein levels and show extensive apoptotic cell death throughout the somatosensory cortex. Furthermore, alpha7 L250T nAChRs are functionally expressed on neurons within the brains of T/T neonatal mice and have properties that are consistent with those observed for the rat alpha7 L250T and the chick alpha7 L247T mutant nAChRs expressed in oocytes. These findings indicate that neurons in the developing brain expressing only alpha7 L250T mutant nAChRs are susceptible to abnormal apoptosis, possibly due to increased Ca2+ influx. PMID- 10800962 TI - Involvement of phosphatidylcholine-specific phospholipase C in thromboxane A2 induced activation of mitogen-activated protein kinase in astrocytoma cells. AB - Thromboxane A2 (TXA2) receptor-mediated signal transduction was investigated in 1321N1 human astrocytoma cells. 9,11-Epithio-11,12-methano-TXA2 (STA2), a TXA2 receptor agonist, induced Ca2+ mobilization and phosphoinositide hydrolysis in a concentration-dependent manner. These responses were inhibited by treatment with U73122, an inhibitor of phosphatidylinositol-specific phospholipase C, or by culturing in 0.5% fetal calf serum containing 0.5 mM dibutyryladenosine 3',5' cyclic monophosphate (dbcAMP) for 2 days. However, the dbcAMP treatment augmented the TXA2 receptor-mediated phosphorylation of mitogen-activated protein kinase (MAPK). These results were confirmed by a functional MAPK assay measuring the incorporation of 32P into the MAPK substrate peptide. The TXA2 receptor-mediated MAPK activation was inhibited by SQ29548, a TXA2 receptor antagonist, and GF109203X, an inhibitor of protein kinase C. Although U73122 did not inhibit or only slightly inhibited the activation of MAPK, D-609, an inhibitor of phosphatidylcholine-specific phospholipase C, potently attenuated the activation in a concentration-dependent manner. Furthermore, STA2 accelerated the release of [3H]choline metabolites from the cells prelabeled with [3H]choline chloride. This release was inhibited by treatment with D-609. These results suggest that phosphatidylcholine-specific phospholipase C and protein kinase C, but not phosphatidylinositol-specific phospholipase C, are involved in TXA2 receptor mediated MAPK activation in 1321N1 human astrocytoma cells. PMID- 10800963 TI - Azido- and isothiocyanato-substituted aryl pyrazoles bind covalently to the CB1 cannabinoid receptor and impair signal transduction. AB - 3-Azidophenyl- and 3-isothiocyanatophenyl-and 2-(5'-azidopentyl)- and 2-(5' isothiocyanatopentyl)pyrazoles were synthesized to determine whether these compounds could behave as covalently binding ligands for the CB1 cannabinoid receptor in rat brain membranes. Heterologous displacement of [3H]CP55940 indicated that the apparent affinity of these compounds for the CB1 receptor was similar to that of the parent compound, SR141716A, with the exception of the 3 isothiocyanato derivatives, which showed a 10-fold loss of affinity. The 3 azidophenyl and 3-isothiocyanatophenyl compounds behaved as antagonists against the cannabinoid agonist desacetyllevonantradol in activation of G proteins [guanosine 5'-O-(y-[35S]thio)triphosphate ([35S]GTPgammaS) binding] and regulation of adenylyl cyclase. The 2-(5'-azidopentyl)- and 2-(5' isothiocyanatopentyl)pyrazoles were poor antagonists for [35S]GTPgammaS binding, and both compounds failed to antagonize the cannabinoid regulation of adenylyl cyclase. After incubation with the isothiocyanato analogues or UV irradiation of the azido analogues, the 3-substituted aryl pyrazoles formed covalent bonds with the CB1 receptor as evidenced by the loss of specific binding of [3H]CP55940. In the case of the isothiocyanato analogues, the log concentration-response curve for cannabinoid-stimulated [35S]GTPgammaS binding was shifted to the right, indicating that loss of receptors compromised signal transduction capability. These irreversibly binding antagonists might be useful tools for the investigation of tolerance and receptor down-regulation in both in vitro and in vivo studies. PMID- 10800964 TI - Characterization of a receptor for insect tachykinin-like peptide agonists by functional expression in a stable Drosophila Schneider 2 cell line. AB - STKR is an insect G protein-coupled receptor, cloned from the stable fly Stomoxys calcitrans. It displays sequence similarity to vertebrate tachykinin [or neurokinin (NK)] receptors. Functional expression of the cloned STKR cDNA was obtained in cultured Drosophila melanogaster Schneider 2 (S2) cells. Insect tachykinin-like peptides or "insectatachykinins," such as Locusta tachykinin (Lom TK) III, produced dose-dependent calcium responses in stably transfected S2-STKR cells. Vertebrate tachykinins (or neurokinins) did not evoke any effect at concentrations up to 10(-5) M, but an antagonist of mammalian neurokinin receptors, spantide II, inhibited the Lom-TK III-induced calcium response. Further analysis showed that the agonist-induced intracellular release of calcium ions was not affected by pretreatment of the cells with pertussis toxin. The calcium rise was blocked by the phospholipase C inhibitor U73122. In addition, Lom-TK III was shown to have a stimulatory effect on the accumulation of both inositol 1,4,5-trisphosphate and cyclic AMP. These are the same second messengers that are induced in mammalian neurokinin-dependent signaling processes. PMID- 10800965 TI - Modulation of GABA(A) receptor subunit mRNA levels in olivocerebellar neurons of purkinje cell degeneration and weaver mutant mice. AB - In olivocerebellar circuits, changes in the subunit composition of GABA(A) receptors occur at a time of extensive synaptic remodeling. In the deep cerebellar nuclei, GABA(A) receptor alpha1, beta2, and gamma2 subunit mRNA expression increases throughout neonatal development, whereas in the inferior olivary complex, the perinatal combination of alpha3, alpha5, beta3, and gamma2 mRNAs switches to the adult combination of alpha2, alpha4, beta3 and gamma1 during postnatal week 2. In situ hybridization was used to examine changes in subunit expression in the olivocerebellar nuclei of Purkinje cell degeneration and weaver mutant mice. In Purkinje cell degeneration, subunit transcripts decreased below control levels in olivary neurons; however, alpha1, beta2, and gamma2 transcript levels were slightly increased in the medial nucleus of the deep cerebellar nuclei. In weaver olivary neurons, although the switch from early to late-onset subunit mRNAs occurred as in normal mice, transcript levels were differentially modulated by the mutation. Our studies indicate that major alterations in synaptic connectivity do not prevent developmentally programmed switches in GABA(A) receptor gene expression but can modulate the timing and level of transcript expression in afferent and efferent neurons. PMID- 10800966 TI - Characterization of arginine decarboxylase in rat brain and liver: distinction from ornithine decarboxylase. AB - We compared the properties of mammalian arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) in rat liver and brain. Mammalian ADC is thermally unstable and associated with mitochondrial membranes. ADC decarboxylates both arginine (Km = 0.75 mM) and ornithine (Km = 0.25 mM), a reaction not inhibited by the specific ODC inhibitor, difluoromethylomithine. ADC activity is inhibited by Ca2+, Co2+, and polyamines, is present in many organs being highest in aorta and lowest in testis, and is not recognized by a specific monoclonal antibody to ODC. In contrast, ODC is thermally stable, cytosolic, and mitochondrial and is expressed at low levels in most organs except testis. Although ADC and ODC are expressed in cultured rat C6 glioma cells, the patterns of expression during growth and confluence are very different. We conclude that mammalian ADC differs from ADC isoforms expressed in plants, bacteria, or Caenorhabditis elegans and is distinct from ODC. ADC serves to synthesize agmatine in proximity to mitochondria, an organelle also harboring agmatine's degradative enzyme, agmatinase, and a class of imidazoline receptor (I2) to which agmatine binds with high affinity. PMID- 10800967 TI - Charge alterations of E22 enhance the pathogenic properties of the amyloid beta protein. AB - Cerebral amyloid angiopathy (CAA) due to amyloid beta-protein (Abeta) is a key pathological feature of patients with Alzheimer's disease and hereditary cerebral hemorrhage with amyloidosis, Dutch-type (HCHWA-D). The CAA in these disorders is characterized by deposition of Abeta in the smooth muscle cells within the cerebral vessel wall. Recently, a new mutation in Abeta, E22K, was identified in several Italian families that, like HCHWA-D, is associated with CAA and hemorrhagic stroke. These two similar disorders, stemming from amino acid substitutions at position 22 of Abeta, implicate the importance of this site in the pathology of HCHWA. Previously we showed that HCHWA-D Abeta(1-40) containing the E22Q substitution induces robust pathologic responses in cultured human cerebrovascular smooth muscle cells (HCSM cells), including highly elevated levels of cell-associated Abeta precursor (AbetaPP) and cell death. In the present study, a series of E22 mutant Abeta(1-40) peptides were synthesized, and their pathogenic properties toward cultured HCSM cells were evaluated. Quantitative fluorescence analyses showed that mutant Abeta(1-40) peptides either containing a loss of charge (E22Q and E22A) or a change of charge (E22K) bind to the surface of HCSM cells and form amyloid fibrils. Similarly, this same group of E22 mutant Abeta(1-40) peptides caused enhanced pathologic responses in HCSM cells. In contrast, wild-type E22 or the charge-preserving E22D Abeta(1-40) peptides were devoid of any of these pathogenic properties. These data suggest that a change or loss of charge at position 22 of Abeta enhances the pathogenic effects of the peptide toward HCSM cells and may contribute to the pathogenesis of the phenotypically related HCHWA disorders. PMID- 10800968 TI - Deficiency of inducible nitric oxide synthase protects against MPTP toxicity in vivo. AB - MPTP produces clinical, biochemical, and neuropathologic changes reminiscent of those that occur in idiopathic Parkinson's disease (PD). In the present study we show that MPTP treatment led to activation of microglia in the substantia nigra pars compacta (SNpc), which was associated and colocalized with an increase in inducible nitric oxide synthase (iNOS) expression. In iNOS-deficient mice the increase of iNOS expression but not the activation of microglia was blocked. Dopaminergic SNpc neurons of iNOS-deficient mice were almost completely protected from MPTP toxicity in a chronic paradigm of MPTP toxicity. Because the MPTP induced decrease in striatal concentrations of dopamine and its metabolites did not differ between iNOS-deficient mice and their wild-type littermates, this protection was not associated with a preservation of nigrostriatal terminals. Our results suggest that iNOS-derived nitric oxide produced in microglia plays an important role in the death of dopaminergic neurons but that other mechanisms contribute to the loss of dopaminergic terminals in MPTP neurotoxicity. We conclude that inhibition of iNOS may be a promising target for the treatment of PD. PMID- 10800969 TI - Analysis of VMAT2 binding after methamphetamine or MPTP treatment: disparity between homogenates and vesicle preparations. AB - [3H]Dihydrotetrabenazine ([3H]DTBZ), a specific ligand for the vesicular monoamine transporter (VMAT2), has been used to characterize the integrity of monoaminergic nerve terminals in experimental animals and humans. The purpose of the present studies was to compare the loss of VMAT2 binding with the loss of other neurochemical markers of the dopamine (DA) nerve terminals in mice treated with neurotoxic doses of methamphetamine (METH) or MPTP. Profound decreases (> or =70%) in DA content, tyrosine hydroxylase activity, and PH]carbomethoxy-3-(4 fluorophenyl)tropane binding to the DA transporter were observed in striatal homogenates at both 1 and 6 days after exposure to the neurotoxins. It is surprising that no significant loss of [3H]DTBZ binding in the homogenates was observed at 1 day after exposure, although a significant loss (-50%) was apparent 6 days later. However, in isolated vesicle preparations, [3H]DTBZ binding and active [3H]DA uptake were markedly reduced (>70%) at 1 day. These observations indicate that vesicle function is compromised at an early time point after exposure to neurotoxic insult. Furthermore, the changes in [H]DTBZ binding in homogenates may not be a sensitive indicator of early damage to synaptic vesicles, although homogenate binding reliably identifies a loss of VMAT2 at later times. PMID- 10800971 TI - Efficacy, effectiveness, and sustainability: translating research into improvements in health care. PMID- 10800970 TI - Methamphetamine rapidly decreases vesicular dopamine uptake. AB - Vesicular sequestration is important in the regulation of cytoplasmic concentrations of monoamines such as dopamine. Moreover, recent evidence suggests that increases in cytoplasmic dopamine levels, perhaps attributable to changes in vesicular monoamine transporter function, contribute to methamphetamine-induced dopaminergic deficits. Hence, we examined whether striatal vesicular uptake is altered following methamphetamine treatment. Multiple administrations of methamphetamine rapidly (within 1 h) decreased vesicular dopamine uptake and dihydrotetrabenazine binding, an effect that (a) persisted at least 24 h, (b) was associated with dopamine and not serotonin neurons, and (c) was unrelated to residual drug introduced by the original methamphetamine treatment. These data suggest that methamphetamine rapidly decreases vesicular monoamine transporter function in dopaminergic neurons, a phenomenon that may be associated with the long-term damage caused by this stimulant. PMID- 10800972 TI - Implementation and effectiveness of a brief smoking-cessation intervention for hospital patients. AB - OBJECTIVES: Previous research has documented that hospital-based smoking cessation counseling is efficacious and cost-effective when delivered by research staff. This study evaluated the implementation and effectiveness of this intervention program when delivered by respiratory therapists chosen from the regular hospital staff. METHODS: A total of 1,173 hospitalized smokers were randomly assigned to either usual care or a stage-based bedside counseling program supplemented with a videotape, self-help materials, and a follow-up telephone call. RESULTS: Using an intent-to-treat analysis and counting those lost to follow-up as smokers, we did not find a significant difference in outcome between intervention (14.2% reported being abstinent for > or =6 months at the 1 year follow-up) and usual care conditions (13.6% abstinence). Process analyses revealed that these results were due to a combination of failure to reach many patients and reduced effectiveness of respiratory therapist interventionists compared with experienced professional counselors in a previous study conducted in the same hospitals. CONCLUSIONS: We recommend implementation of hospital-based smoking-cessation counseling by professional counselors whose primary responsibility is to deliver the intervention. Recommendations for future research and for innovative ways to reach hospitalized smokers who are not receiving intervention are discussed. PMID- 10800973 TI - Building uncertainty into cost-effectiveness rankings: portfolio risk-return tradeoffs and implications for decision rules. AB - BACKGROUND: Current principles of cost-effectiveness analysis emphasize the rank ordering of programs by expected economic return (eg, quality-adjusted life-years gained per dollar expended). This criterion ignores the variance associated with the cost-effectiveness of a program, yet variance is a common measure of risk when financial investment options are appraised. Variation in health care program return is likely to be a criterion of program selection for health care managers with fixed budgets and outcome performance targets. METHODS: Characterizing health care resource allocation as a risky investment problem, we show how concepts of portfolio analysis from financial economics can be adopted as a conceptual framework for presenting cost-effectiveness data from multiple programs as mean-variance data. RESULTS: Two specific propositions emerge: (1) the current convention of ranking programs by expected return is a special case of the portfolio selection problem in which the decision maker is assumed to be indifferent to risk, and (2) for risk-averse decision makers, the degree of joint risk or covariation in cost-effectiveness between programs will create incentives to diversify an investment portfolio. CONCLUSIONS: The conventional normative assumption of risk neutrality for social-level public investment decisions does not apply to a large number of health care resource allocation decisions in which health care managers seek to maximize returns subject to budget constraints and performance targets. Portfolio theory offers a useful framework for studying mean variance tradeoffs in cost-effectiveness and offers some positive predictions (and explanations) of actual decision making in the health care sector. PMID- 10800974 TI - Consumer responses to health plan report cards in two markets. AB - BACKGROUND: Health plans can compete on quality when consumers have helpful information. Report cards strive to meet this need, but consumer responses have not been measured. OBJECTIVES: The objectives of this study were (1) to compare consumer responses to report cards in 2 markets, (2) to determine how personal characteristics relate to exposure, and (3) to assess the perceived helpfulness of the report cards. RESEARCH DESIGN: A postenrollment survey was used. SUBJECTS: The study included 784 employees of Monsanto (St Louis, 1996) and 670 employees of a health care purchasing cooperative (Denver, 1997). DEPENDENT MEASURES: The dependent measures were (1) exposure, specifically remembering the report card, and intensity of reading it and (2) perceived helpfulness in learning about plan quality and in deciding to stay or switch. RESULTS: Except for remembering seeing the report card (Denver, 47%; St Louis, 55%), the 2 groups did not differ. Forty percent read most or all of the report card; 82% found the report helpful in learning about quality; and 66% found it helpful in deciding to stay or switch. Employees who used patient survey information in their plan decision were more likely to remember seeing the report card (odds ratio [OR], 4.85), to read it intensely (OR, 2.84), and to find it helpful in learning about plan quality (OR, 3.04) and deciding whether to stay or switch plans (OR, 2.64). CONCLUSIONS: Although the 2 samples differed markedly, their responses to report cards were similar. Exposure and helpfulness were related more to employee preferences for the type of information than to their health care decision needs. PMID- 10800975 TI - Comparison of changes in physical functioning of elderly patients with new diagnoses of cancer. AB - BACKGROUND: Controversy surrounds the impact of site of cancer and treatments on functioning of elderly cancer patients. OBJECTIVES: This research determines (1) whether age, gender, comorbid conditions, site and stage of cancer, and treatments are related to losses in physical functioning at 4 observations during the year after diagnosis; (2) whether symptoms are a mediating variable between treatment and function; and (3) which indicators account for true change in functioning in the year after diagnosis. METHODS: An inception cohort of 907 patients aged > or =65 years and newly diagnosed with breast, colon, lung, or prostate cancer were accrued from 24 community oncology programs. Stage and treatment data were obtained from medical records. Physical functioning was measured with the SF-36 subscale. Interviews were conducted at 6 to 8, 12 to 16, 26 to 30, and 52 weeks after diagnosis. RESULTS: Men scored 10 points higher on physical function than women at all observation points. Patients with > or =3 comorbid conditions scored lower in functioning. Interactions between site of cancer and treatment modalities were observed. Pain, fatigue, and numbers of symptoms were independent predictors of loss of function. Surgery, female gender, and number of symptoms predicted reliable change in function. CONCLUSIONS: Elderly patients with cancer report levels of function similar to other chronic conditions. Scores on physical function varied by site of cancer; the pattern of change was similar among sites. Age, comorbidity, treatment modalities, and symptom reports each had an independent effect on loss of functioning. Untreated breast cancer patients had lower functioning, suggesting a possible treatment bias. PMID- 10800976 TI - Posthospitalization home health care use and changes in functional status in a Medicare population. AB - OBJECTIVE: The objective of this work was to estimate the effect of Medicare beneficiaries' use of home health care (HHC) for 6 months after hospital discharge on the change in functional status over a 1-year period beginning before hospitalization. DATA SOURCES AND STUDY SETTING: Data came from the Medicare Current Beneficiary Survey, which is a nationally representative sample of Medicare beneficiaries, in-person interview data, and Medicare claims for 1991 through 1994 for 2,127 nondisabled, community-dwelling, elderly Medicare beneficiaries who were hospitalized within 6 months of their annual in-person interviews. STUDY DESIGN: Econometric estimation with the instrumental variable method was used to correct for observational data bias, ie, the nonrandom allocation of discharged beneficiaries to the use of posthospitalization HHC. The analysis estimates a first-stage model of HHC use from which an instrumental variable estimate is constructed to estimate the effect on change in functional status. PRINCIPAL FINDINGS: The instrumental variable estimates suggest that HHC users experienced greater improvements in functional status than nonusers as measured by the change in a continuous scale based on the number and mix of activities of daily living and instrumental activities of daily living before and after hospitalization. The estimated improvement in functional status could be as large as 13% for a 10% increase in HHC use. In contrast, estimation with the observational data on HHC use implies that HHC users had poorer health outcomes. CONCLUSIONS: Adjusting for potential observational data bias is critical to obtaining estimates of the relationship between the use of posthospitalization HHC and the change in health before and after hospitalization. After adjustment, the results suggest that efforts to constrain Medicare's spending for HHC, as required by the Balanced Budget Act of 1997, may lead to poorer health outcomes for some beneficiaries. PMID- 10800977 TI - Socioeconomic status and dissatisfaction among HMO enrollees. AB - OBJECTIVES: Member satisfaction is commonly used as an indicator of the quality of care delivered by health plans. Yet few contemporary studies have explored the extent to which individual patient characteristics influence dissatisfaction in HMOs. We sought to determine whether socioeconomic status is associated with enrollee dissatisfaction. METHODS: Data are from a cross-sectional, telephone survey of a probability sample of adults enrolled in New Jersey HMOs in 1998 (n = 7,983). Health plan ratings were elicited as part of the Consumer Assessment of Health Plans Study (CAHPS) survey, along with income, education, and race/ethnicity. Other factors known to influence satisfaction (age, gender, health status, extent of plan choice, and payment for plan) were also ascertained. RESULTS: Socioeconomically advantaged enrollees were more likely to give low ratings to their health plans. In a multivariate logistic regression model, those with incomes exceeding $100,000 had 1.65 times the odds of being dissatisfied compared with those with family incomes less than $25,000 (P <0.001); those with a college education had 2.53 times the odds of being dissatisfied than those who had not completed high school (P <0.001). However, among enrollees in their plans for > or =5 years, those in the lowest income group were significantly more dissatisfied than higher-income enrollees. CONCLUSIONS: Among New Jersey HMO enrollees, higher socioeconomic status (SES) is associated with greater dissatisfaction. Although based on cross-sectional data and thus preliminary, the evidence presented here also suggests that the SES dissatisfaction relationship varies as a function of duration of enrollment. Further research using longitudinal data could shed additional light on the SES dissatisfaction link. PMID- 10800979 TI - Impact of individual and market factors on the timing of initiation of hospice terminal care. AB - CONTEXT: Hospice terminal care is now used by 10% to 15% of elderly Americans at variable points before their deaths. OBJECTIVE: By examining the duration of patient survival after enrollment in hospice care, we sought to identify individual and market factors associated with the timing of hospice use. DESIGN: We linked Medicare claims, census information, and Area Resource File data to form a national cohort of 151,410 hospice patients admitted in 1993 and followed up until late 1996. We examined this cohort with Cox regression and other means. MAIN OUTCOME MEASURE: The primary outcome measure was survival after hospice enrollment. RESULTS: The patients had a mean +/- SD age of 79.0 +/- 7.4 years; 10.2% were nonwhite; 51.4% were female; and 71.3% had cancer. Median survival after hospice enrollment was 30 days (interquartile range, 10-86 days). After adjustment for measured patient, provider, and market factors, several variables were associated with relatively earlier hospice enrollment, farther from death. Compared with complementary groups, nonwhites were enrolled in hospice 4 days earlier; women, 5 days earlier; older people, 1 day earlier; and those with substance abuse, psychiatric disease, or dementia, each 3 days earlier. After adjustment, income and education were not associated with the timing of enrollment. Patients residing in markets with more hospital beds, greater hospice capacity, or a higher proportion of generalists were enrolled earlier. CONCLUSIONS: Even after adjustment for certain clinical attributes, individual social factors and local market factors were associated with survival after hospice enrollment. Certain socially disadvantaged groups were enrolled earlier, as were those residing in areas with more medical institutions. The decision to enroll patients in hospice may depend on both nonclinical and clinical factors. PMID- 10800978 TI - Evaluating the equivalence of health care ratings by whites and Hispanics. AB - PURPOSE: This study was designed to assess the equivalence of a health care ratings scale administered to non-Hispanic white and Hispanic survey respondents. METHODS: We sent 18,840 questionnaires to a random sample of patients receiving medical care from a physician group association concentrated in the western United States; 7,093 were returned (59% adjusted response rate). Approximately 90% of survey respondents self-identified as white/Caucasian (n = 5,508) or Hispanic/Latino (n = 713). Interpersonal and technical aspects of medical care were assessed with 9 items, all administered with a 7-point response format: the best, excellent, very good, good, fair, poor, and very poor, with a "not applicable" option. Item response theory procedures were used to test for differential item functioning between white and Hispanic respondents. RESULTS: Hispanics were found to be significantly more dissatisfied with care than whites (effect size=0.27; P <0.05). Of the 9 test items, 2 had statistically significant differential item functioning (P <0.05): reassurance and support offered by your doctors and staff and quality of examinations received. However, summative scale scores and test characteristic curves for whites and Hispanics were similar whether or not these items were included in the scale. CONCLUSIONS: Despite some differences in item functioning, valid satisfaction-with-care comparisons between whites and Hispanics are possible. Thus, disparities in satisfaction ratings between whites and Hispanics should not be ascribed to measurement bias but should be viewed as arising from actual differences in experiences with care. PMID- 10800980 TI - SF-36 health survey: tests of data quality, scaling assumptions, and reliability in a community sample of Chinese Americans. AB - BACKGROUND: Chinese Americans are one of the fastest growing ethnic groups in the United States; however, language and cultural obstacles have challenged health workers and policy makers seeking to understand the health status and needs of this population. OBJECTIVES: This study is the first to use a large-scale probability design to evaluate the 36-item Short-Form Health Survey (SF-36) in a Chinese population (n = 1,501). METHODS: Using the International Quality of Life Assessment Project protocols, we examine summated-rating scaling assumptions, item-internal consistency, item-discriminant validity, and reliability. RESULTS: Similar to previous studies, our tests indicated that the SF-36 generally met minimum psychometric criteria with high reliability and satisfactory scaling success rates for most scales. However, the performance of the vitality and mental health scales was less satisfactory with regard to discriminant validity and scaling success rates. Notably, our results indicate that VT3 and VT4 ("feel worn out" and "tired", respectively) formed a separate "fatigue" cluster more highly correlated with the mental health scale. However, MH4 and MH5 ("downhearted and blue" [reverse coded] and "been a happy person") were more highly correlated with the vitality scale, suggesting that it may be more meaningful to reorganize the vitality and mental health items along the dimensions of well-being and distress. CONCLUSIONS: These results are interpreted within a cultural framework; however, additional work is needed to better understand the relationship between vitality and mental health for Chinese Americans. PMID- 10800981 TI - Malignant lymphoma of the thyroid gland: a clinicopathologic study of 108 cases. AB - We report a retrospective clinicopathologic study of 108 primary thyroid gland lymphomas (PTLs), classified using the REAL and proposed WHO classification schemes. The patients included 79 women and 29 men, with an average age of 64.3 years. All patients presented with a thyroid mass. The PTLs were classified as marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) or MZBL (n = 30), diffuse large B-cell lymphoma (DLBCL) with MZBL (n = 36), DLBCL without MZBL (n = 41), and follicle center lymphoma (FCL; n = 1). Excluding the FCL, features of lymphomas of MALT-type were identified in all groups, despite a follicular architecture in 23% of cases. Lymphocytic thyroiditis (LT) was identified in 94%. Ninety-one percent of patients presented with stage IE or IIE disease, whereas 69% had perithyroidal soft tissue infiltration. All patients were treated with surgical excision followed by adjuvant therapy (76%): chemotherapy (15%), radiation (19%), or a combination of radiation and chemotherapy (42%). Disease-specific survival was 82% at last follow up (mean, 82.8 mos) and 79% at 5 years. Statistically, stages greater than IE, presence of DLBCL, rapid clinical growth, abundant apoptosis, presence of vascular invasion, high mitotic rate, and infiltration of the perithyroidal soft tissue were significantly associated with death with disease. No patients with MZBL or stage IE disease died with disease. In summary, PTLs typically occur in middle- to older-aged individuals as a thyroid mass, with a predilection for females. Despite their histologic heterogeneity and frequent simulation of other lymphoma subtypes, virtually all PTLs are lymphomas of MALT-type arising in the setting of LT. Mixed DLBCL and MZBL are common. Overall, PTLs have a favorable outcome with appropriate therapy, but prognosis depends on both clinical stage and histology. MZBL and stage IE tumors have an excellent prognosis, whereas tumors with a large cell component or DLBCL or stage greater than IE have the greatest potential for a poor outcome. PMID- 10800982 TI - Extracranial sinonasal tract meningiomas: a clinicopathologic study of 30 cases with a review of the literature. AB - Extracranial meningiomas of the sinonasal tract are rare tumors. These tumors are frequently misclassified, resulting in inappropriate clinical management. To date, there has been no comprehensive study to evaluate the clinicopathologic aspects of meningioma in these anatomic sites. Thirty cases of sinonasal tract meningiomas diagnosed between 1970 and 1992 were retrieved from the files of the Otorhinolaryngic Registry of the AFIP. Histologic features were reviewed, immunohistochemical studies were performed, patient follow up was obtained, and the results were statistically analyzed. The patients included 15 females and 15 males, aged 13 to 88 years (mean, 47.6 yrs). Patients presented clinically with a mass, epistaxis, sinusitis, pain, visual changes, or nasal obstruction, dependent on the anatomic site of involvement. Symptoms were present for an average of 31.1 months. The tumors affected the nasal cavity (n = 14), nasopharynx (n = 3), frontal sinus (n = 2), sphenoid sinus (n = 2). or a combination of the nasal cavity and ethmoid, frontal, sphenoid, and/or maxillary sinuses (n = 9). The tumors ranged in size from 1.0 to 8.0 cm in greatest dimension (mean, 3.5 cm). Radiographic studies demonstrated a central nervous system connection in six cases. The tumors often eroded the bones of the sinuses (n = 18) and involved the surrounding soft tissues, the orbit, and occasionally the base of the skull. Histologically, the tumors demonstrated features similar to intracranial meningiomas. The majority were of the meningothelial type (n = 23), although there were three atypical meningiomas. Immunohistochemical studies confirmed the diagnosis of meningioma with positive reactions for epithelial membrane antigen (EMA) and vimentin (all tested). The differential diagnosis includes paraganglioma, carcinoma, melanoma, psammomatoid ossifying fibroma, and angiofibroma. Surgical excision was used in all patients. Three patients died with recurrent disease (mean, 1.2 yrs), one was alive with recurrent disease (25.6 years), and the remaining 24 patients were alive or had died of unrelated causes (mean, 13.9 yrs) at the time of last follow up (two patients were lost to follow up). Extracranial sinonasal tract meningiomas are rare tumors which need to be considered in the differential diagnosis of sinonasal tumors. A whorled growth pattern and psammoma bodies, combined with positive EMA and vimentin immunohistochemical reactions, can confirm the diagnosis of meningioma. The overall prognosis is good, without a difference in outcome between benign and atypical meningiomas. PMID- 10800983 TI - Pathologists' agreement with experts and reproducibility of breast ductal carcinoma-in-situ classification schemes. AB - Several histologic classifications for breast ductal carcinoma in situ (DCIS) have been proposed. This study assessed the diagnostic agreement and reproducibility of three DCIS classifications (Holland [HL], modified Lagios [LA], and Van Nuys [VN]) by comparing the interpretations of pathologists without expertise in breast pathology with those of three breast pathology experts, each a proponent of one classification. Seven nonexpert pathologists in New Hampshire and three experts evaluated 40 slides of DCIS according to the three classifications. Twenty slides were reinterpreted by each nonexpert pathologist. Diagnostic accuracy (nonexperts compared with experts) and reproducibility were evaluated using inter- and intrarater techniques (kappa statistic). Final DCIS grade and nuclear grade were reported most accurately among nonexpert pathologists using HL (kappa = 0.53 and 0.49, respectively) compared with LA and VN (kappa = 0.29 and 0.35, respectively, for both classifications). An intermediate DCIS grade was assessed most accurately using HL and LA, and a high grade (group 3) was assessed most accurately using VN. Diagnostic reproducibility was highest using HL (kappa = 0.49). The VN interpretation of necrosis (present or absent) was reported more accurately than the LA criteria (extensive, focal, or absent; kappa = 0.59 and 0.45, respectively), but reproducibility of each was comparable (kappa = 0.48 and 0.46, respectively). Intrarater agreement was high overall. Comparing all three classifications, final DCIS grade was reported best using HL. Nuclear grade (cytodifferentiation) using HL and the presence or absence of necrosis were the criteria diagnosed most accurately and reproducibly. Establishing one internationally approved set of interpretive definitions, with acceptable accuracy and reproducibility among both pathologists with and without expertise in breast pathology interpretation, will assist researchers in evaluating treatment effectiveness and characterizing the natural history of DCIS breast lesions. PMID- 10800984 TI - Smooth muscle distribution in the extrahepatic bile duct: histologic and immunohistochemical studies of 122 cases. AB - The distribution of smooth muscle fibers in the extrahepatic bile duct (EBD) wall is not well characterized. We analyzed 101 consecutive Whipple's operation specimens and 21 autopsy specimens for the pattern of smooth muscle distribution in EBD using the Masson-trichrome stain and the desmin immunohistochemical stain. The patterns were categorized as continuous, interrupted, scattered, and no muscle layer. EBDs were divided into lower, middle, and upper portions, and the distribution pattern of smooth muscle fibers was analyzed separately in each portion. Because most surgically resected specimens contained the middle and lower EBDs with only a portion of the upper EBD, only the length of the middle and lower EBDs (common bile duct, CBD) was measured. The mean length of CBD in surgically resected specimens was 6.4 +/- 1.4 cm (men, 6.6 +/- 1.3 cm; women, 6.1 +/- 1.5 cm). The mean length of CBD in autopsy specimens was 6.8 +/- 1.0 cm. The predominant patterns of the lower third of the EBD were interrupted (49%) and continuous (43%). The predominant patterns of the middle third of the EBD were scattered (63%) and interrupted (23%). Those of the upper third of the EBD were no muscle fiber (58%) and scattered (39%). In conclusion, different patterns of smooth muscle distribution were observed in different portions of the EBD. Because scattered muscle fibers or no muscle fibers were the main features of the upper third of the EBD, understanding of this pattern may be helpful for assessment of the depth of invasion or staging of carcinoma of the upper third of the EBD. PMID- 10800985 TI - Crohn's colitis-like changes in sigmoid diverticulitis specimens is usually an idiosyncratic inflammatory response to the diverticulosis rather than Crohn's colitis. AB - The clinical outcome and optimum classification of patients who have sigmoid resection specimens that show the histologic features of Crohn's disease (CD) and diverticulitis is not well defined. Historically, these patients were considered to have coexistent diseases, but recent studies have suggested that the CD-like changes are part of the inflammatory reaction of the diverticulitis. Sorting out these issues has been complicated by the lack of distinction between patients with and without CD in other regions of the bowel, short clinical follow-up periods, and small numbers of patients. We report on the clinical outcome and histology of 29 patients who had sigmoid resection specimens with diverticulitis and CD-like changes. Of the 25 patients who had no prior or concurrent CD at the time of surgery, 23 remained free of CD during the follow-up period (median, 6.0 yrs) and two developed CD in other regions of the bowel. All four patients with CD prior to their sigmoid resection continued to have active CD postoperatively. There were no histologic features of the sigmoid resection specimens that could be associated with the outcome of the patient. These results suggest that CD-like changes within the sigmoid resection specimen are an idiosyncratic inflammatory response to the diverticulosis rather than coexistent CD in the overwhelming majority of patients who do not have prior or concurrent CD at the time of sigmoid resection. Pathologists should be wary about making the diagnosis of sigmoid CD in the context of diverticulitis unless there is CD in other parts of the bowel. PMID- 10800986 TI - The histologic spectrum and clinical outcome of refractory and unclassified sprue. AB - The vast majority of patients with celiac disease respond to a gluten-free diet; yet, a small number of refractory patients do not respond and have persistent malabsorption and residual mucosal abnormalities of the small intestine. The histologic features of refractory/unclassified sprue have been published as case reports, often without long-term follow up, and no clear histologic picture has emerged. We present the results of a long-term study of the clinical and histologic features of 10 patients with refractory/unclassified sprue. The histologic features of small bowel biopsies in this group of patients were compared with those of 10 patients with responsive celiac disease and with 10 patients without malabsorption who had normal duodenal biopsies. Five of the 10 refractory patients ultimately developed collagenous sprue as a distinct histologic marker of refractory disease. Additional distinctive findings found in small bowel biopsies in the refractory group were subcryptal chronic inflammation (10 of 10) and marked mucosal thinning in three patients. Other nonspecific findings included acute inflammation and gastric metaplasia. One patient with collagenous sprue developed a B-cell lymphoma of the ileum, and in general collagenous sprue was associated with a poor prognosis. Two of five patients died whereas two others require total parenteral nutrition for survival. Pathologists evaluating small bowel biopsies in the setting of malabsorption should be aware of the subtle histologic changes described here that may portend a refractory course. PMID- 10800987 TI - Increased incidence of follicular lymphoma in the duodenum. AB - The incidence of indolent lymphomas in the lymph nodes and extranodal regions is quite different. Follicular lymphoma (FL) is most common in the nodes, and it seems to be least common in the gastrointestinal (GI) tract, where mucosa associated lymphoid tissue lymphoma arises most frequently. The authors report that the incidence of FL is unexpectedly high in the duodenum compared with other portions of the GI tract. FL was detected in only eight of 222 cases of GI lymphoma (3.6%). However, five cases of FL arose in the duodenum, which accounted for 38.5% of 13 duodenal lymphomas. Only in two patients did FL arise in either the stomach or the colorectum, and in the remaining patients FL was widespread with lymphomatous polyposis. Duodenal FL was composed of neoplastic follicles with small cleaved cells in dominance, and the immunophenotype of the lymphoma cells was CD10+, BCL-2+, CD20+, CD75+, CD79+, CD3-, CD5-, cyclin D1-, CD23-, and CD45RO-. All the patients were women age 37 to 66 years (average age, 52.4 yrs). In all patients the lymphoma was present around the ampulla of Vater, and four of five patients showed multiple small-size polyps. Although lymphoma cell infiltration was confined to the submucosa in the four patients examined, the regional lymph nodes were involved partially in two patients without distant metastasis. All patients are alive at 2 to 50 months of follow up (average, 27 mos), which is comparable with the prognosis for indolent nodal lymphomas. These results suggest that the duodenum has a distinct background of histogenesis of the lymphomas and that biopsy specimens from the duodenum with multiple polyps should be examined carefully. PMID- 10800988 TI - Clinicopathologic reassessment of primary cutaneous B-cell lymphomas with immunophenotypic and molecular genetic characterization. AB - Primary cutaneous B-cell lymphomas (PCBLs) may have particular clinicopathologic characteristics distinct from their lymph node-based counterparts. It has been suggested that PCBLs should have a separate classification system. The aim of this study was to determine whether the Revised European-American Lymphoid Neoplasms (REAL) classification is applicable to PCBL. Thirty-nine cases of PCBL from 36 patients, consisting of 20 men and 16 women (median age 66 yrs), were included in this study. Paraffin-section immunohistochemistry for CD3, CD5, CD10, CD20, CD43, Bcl-2, Bcl-6, and cyclin D1 was performed in all cases. Immunostaining for immunoglobulin light chains was also performed on cases histologically diagnosed as extranodal marginal zone lymphoma (MZL) and primary cutaneous B-cell lymphoma unclassifiable (PCBLu). Polymerase chain reaction (PCR) analysis of t(14;18) was performed in all cases. Immunoglobulin heavy chain gene rearrangement (VDJ) was tested by PCR on all follicle center lymphoma (FCL), MZL, and PCBLu cases. The 39 cases consisted of 15 (39%) FCLs, 13 (33%) diffuse large B-cell lymphomas (DLCL), 9 (23%) extranodal MZL, and 2 cases of PCBLu. Anatomically, 59% of PCBLs occurred in the head and neck, of which approximately 57% were FCL. Five of six cases presenting on the lower extremity were DLCL. Follow-up data was available from all 39 patients with a mean of 50.8 months. All but two patients are alive with or without disease at last contact. One patient with DLCL died of lung metastases and the other DLCL patient died of sepsis as a complication of therapy. In all 15 cases of FCL, CD10 and/or Bcl-6 expression supported the follicle center origin of the neoplastic cells. In contrast to previous reports, we found that 53% (8 of 15) of primary cutaneous FCL had either Bcl-2 protein expression or t(14;18). Our data indicate that many cases of primary cutaneous FCL have Bcl-2 alterations similar to their nodal counterpart. We found that 95% (37 of 39) of PCBLs could be classified according to the REAL classification, supporting its applicability in cutaneous lymphomas. PMID- 10800989 TI - Paraffin section immunophenotype of cutaneous and extracutaneous mast cell disease: comparison to other hematopoietic neoplasms. AB - Mast cell disease (MCD) is a rare proliferation that may be easily confused with other hematopoietic tumors. Several paraffin section antibodies immunoreact with mast cells but most are not specific. Tryptase, a specific marker of mast cells, may not be cost-effective to maintain in a laboratory because of the rarity of these lesions. This study was undertaken to assess the immunoreactivity of MCD and attempt to select a limited antibody panel for diagnosing MCD among hematopoietic tumors that morphologically mimic MCD. Immunophenotyping of cutaneous ( 10 cases) and extracutaneous (18 cases) MCD, as well as 94 other hematopoietic neoplasms, was performed on paraffin sections. All cases of MCD showed strong and diffuse positivity for CD68 and tryptase. In the vast majority of the cases, the mast cells were also positive for CD117 (27 of 28) and CD43 (25 of 27). Four cases (40%) of cutaneous MCD demonstrated a subpopulation of mast cells expressing myeloperoxidase (MPX), whereas all extracutaneous MCD were negative for MPX. Two (40%) extramedullary myeloid tumors (EMT) expressed CD43, CD68, CD 117, and MPX, but none expressed tryptase. CD43, CD68, CD117, and tryptase were expressed by 25%, 1%, 15%, and 1%, respectively, of all B-cell lymphoid neoplasms, and none expressed more than one of these four antigens. We conclude that (1) cutaneous MCDs may demonstrate a subpopulation of MPX antigen expressing tumor cells and may be confused with cutaneous involvement by myeloid leukemia if other antibodies are not used; (2) tryptase is the most specific mast cell marker among the antibodies studied; and, (3) the detection of tryptase, together with CD68, CD117, and usually CD43, is unique to MCD among hematopoietic tumors. PMID- 10800990 TI - Some mullerian inclusion cysts in lymph nodes may sometimes be metastases from serous borderline tumors of the ovary. AB - Glandular inclusions that appear morphologically benign are occasionally found in lymph nodes as well as in peritoneal and omental biopsies. In patients with gynecologic malignancies, the nature and significance of these mullerian inclusion cysts (MIC) present a diagnostic challenge with regard to whether they are benign and incidental or are related to the coincident tumor for which surgery is being performed. Sixty-two cases of MIC were prospectively identified during a 6-year period. The frequencies were calculated and stratified by lymph node chain distribution, primary tumor site, and primary tumor type. MIC appeared as small cysts lined by a serous (mullerian)-type, cytologically bland, cuboidal to columnar epithelium with a simple architecture. Among 62 women, MIC was found in lymph nodes (27 cases), pelvic peritoneum (19 cases), omentum (16 cases), bowel serosa (9 cases), uterine serosa (8 cases), and parametrial connective tissues (4 cases). Among a set of 417 consecutive cases in which lymphadenectomy was performed, 46 (11%) women had MIC. The MIC involved multiple sites (26 cases in the peritoneum/omentum and 27 in lymph nodes). The primary tumor was in the ovary in 32 of the 46 women with MIC (70%) and of these, 17 were borderline serous (53%). Sixty-two of 6,154 lymph nodes examined contained MIC (1.0%). 3.2% of nodes contained MIC in which the primary tumor arose in the ovary, but only 0.1% with either endometrial or cervical tumors (chi2, p <0.00001). The lymph nodes most often involved by MIC were from para-aortic sites (40%), which reflect the primary drainage route from the ovary. Not uncommonly, neighboring areas in the same lymph node group with MIC disclosed separate foci of obvious metastatic borderline tumor (4 of 10; 40%). In summary, the increased frequency of MIC in lymph nodes sampled for primary ovarian malignancies suggests that MIC in some cases, rather than being benign, incidental inclusions, are more likely bland appearing forms of metastatic tumor. The preponderance of inclusions occurs with serous ovarian tumors of borderline malignancy, and the inclusions are overrepresented in the lymph nodes that primarily receive drainage from the ovary. PMID- 10800991 TI - Non-Hodgkin's lymphoma involving the vagina: a clinicopathologic analysis of 14 patients. AB - Non-Hodgkin's lymphomas (NHL) uncommonly involve the vagina. In this study, 14 NHL involving the vagina are reported. Eight cases were stage IE or IIE and are presumed to be primary. The mean age of these eight patients at presentation was 42 years (range, 26-66 yrs), and four of eight patients complained primarily of vaginal bleeding. Histologically, all eight neoplasms were diffuse large B-cell lymphoma (DLBCL). Clinical follow up ranged from 1.8 to 18 years. Six of eight patients were alive without evidence of disease at the last follow up (range, 2.8 21 yrs), one patient died of unrelated causes at 9 years, and one patient died from NHL at 1.8 years. In six patients vaginal involvement was part of systemic disease at diagnosis, either stage IIIE or IV. The mean patient age at the time vaginal involvement was detected was 65 years (range, 49-82 yrs). Four of six patients had vaginal bleeding. Five neoplasms were DLBCL and one tumor was B-cell small lymphocytic lymphoma/chronic lymphocytic leukemia. Clinical follow up for these patients ranged from 2 weeks to 13 years. Two patients were free of disease after treatment at 4.5 and 13 years, two patients were alive with progressive NHL, one patient died of NHL, and one patient was recently diagnosed. The authors conclude that low-stage (presumably primary) vaginal NHL are DLBCL, tend to occur in younger women, and cause vaginal bleeding. High-stage NHL involving the vagina are usually DLBCL, tend to affect older women, and are relatively more heterogeneous clinically and histologically, but also usually cause vaginal bleeding. PMID- 10800992 TI - Endometrial intraepithelial carcinoma with associated peritoneal carcinomatosis. AB - Endometrial intraepithelial carcinoma (EIC) is a recently described entity, defined as a noninvasive, cytologically malignant lesion that replaces the endometrial surface epithelium. EIC frequently coexists with uterine serous carcinoma (USC) and is hypothesized to be its precursor lesion. However, the clinical significance and biologic potential of finding EIC without USC is not known. We report three postmenopausal women with EIC alone who were found to have multiple, synchronous foci of extrauterine serous carcinoma at presentation. Because the clinical findings in these patients simulated primary peritoneal serous carcinoma (PSC), we compared the clinicopathologic features of these cases with a group of nine bona fide PSCs for which exhaustively sectioned endometria, fallopian tubes, and ovaries were available for review. The average age of the EIC patients was 73 years. Two patients presented with abdominal distention and one with vaginal bleeding. Hysterectomy in each case showed endometrial polyps with EIC, but without invasive USC, in a background of atrophic endometrium. Bilateral salpingo-oophorectomy and staging showed serous carcinoma involving the ovarian hilum, the surfaces of the fallopian tubes and ovaries, in addition to peritoneal carcinomatosis. p53 overexpression was observed in both EIC and the extrauterine deposits of serous carcinoma in each case. The average age of the PSC patients was 66 years. All nine patients presented with abdominal distention. EIC was not identified in any of the hysterectomy specimens. Bilateral salpingo oophorectomies, omentectomies, and peritoneal biopsies showed peritoneal carcinomatosis, including bulky peritoneal tumor deposits, but only minimal ovarian surface involvement. p53 overexpression was observed in seven cases. These findings indicate that EIC without coincident USC can be associated with invasive, extrauterine serous carcinomatosis. We did not, however, find any significant differences between the clinicopathologic features of primary extrauterine serous carcinomas (PSCs) and those associated with EIC. We conclude that the finding of EIC in an endometrial curettage specimen should prompt a thorough search for an invasive uterine and/or extrauterine serous carcinoma. Conversely, an endometrial origin should be excluded in patients with peritoneal carcinomatosis. PMID- 10800993 TI - The pathology of liver-localized post-transplant lymphoproliferative disease: a report of three cases and a review of the literature. AB - Post-transplantation lymphoproliferative disease (PTLD) is a complication of solid organ transplantation that is typically of B-cell origin and associated with Epstein-Barr virus (EBV). In patients receiving orthotopic liver transplantation (OLT) and treated with cyclosporin A. PTLD typically presents between 6 and 17 months post-transplantation as a systemic illness with involvement of the hepatic graft in a minority of cases. A small number of cases of biopsy-proven PTLD arising in the hepatic graft and limited to the liver and periportal structures have been previously reported. This report describes three additional cases of liver-localized PTLD and reviews similar cases in the literature. The donor/host origin of PTLD may have prognostic significance because the two cases in this report that are of donor origin had different clinical and pathologic features compared with the case of host origin. A rapid PCR-based technique for determining the origin of PTLD is described. PMID- 10800994 TI - CD70 expression in thymic carcinoma. AB - CD70, a type II transmembrane glycoprotein, is a member of the tumor necrosis factor (TNF) family that mediates the interaction between B- and T-lymphocytes. CD70 has been shown to be expressed by malignant lymphoma, especially Hodgkin's disease, and by nasopharyngeal carcinoma, both of which are frequently associated with Epstein-Barr virus (EBV). In this study, we investigated the expression of CD70 in epithelial cells of various types of thymic epithelial tumors and its association with EBV. Immunohistochemical expression of CD70 was studied on frozen tissue. In a series of 27 thymic epithelial tumors, including thymic carcinomas (n = 8), atypical thymomas (n = 5), thymomas (n = 13), and thymic carcinoid (n = 1), 7 (88%) thymic carcinomas and 1 (20%) atypical thymoma showed positive immunoreactivity for CD70, whereas CD70 was not detected in other tumors. Twenty-four intrathoracic malignant epithelial tumors of nonthymic origin, including lung (n = 17), esophagus (n = 5), and mesothelium (n = 2), showed no immunoreactivity for CD70. Northern blot analysis also revealed that CD70 messenger RNA was expressed in 2 of 2 thymic carcinomas, 0 of 2 atypical thymomas. and 0 of 2 thymomas. All of the 27 thymic epithelial tumors were EBV negative as assessed by EBV-encoded small RNA in situ hybridization. The expression of CD70 is closely related to the pathogenesis of thymic carcinoma but unrelated to EBV infection in the thymus. PMID- 10800995 TI - Primary malignant melanoma of the anterior mediastinum in a child. AB - Primary malignant melanoma of the mediastinum is extremely rare. We report a case not previously reported of primary malignant melanoma located in the mediastinum in a 11-year-old boy. The tumor could not be completely resected as a result of extensive invasion of the large blood vessels. Histologically, the tumor was heavily pigmented and composed of vague fascicles of spindle cells intermingled with epithelioid cells. Immunohistochemical analysis showed vimentin, S-100 protein, Melan-A, and HMB-45 immunoreactivity in most of the tumor cells. Nearly 50% of the tumor cells were also positive for p53. It is suggested that primary malignant melanoma of the anterior mediastinum may have a histogenetic relationship to the recently described aggregates of nevus cells in the thymus or mediastinal lymph nodes. PMID- 10800996 TI - Endometrioid neoplasia retrogressive terminology. PMID- 10800997 TI - Lymphocytic and collagenous colitis as possible patterns of Crohn's colitis. PMID- 10800998 TI - Dysplastic nevus. PMID- 10800999 TI - Aircraft control forces and EMG activity in a UH-1H Iroquois helicopter during routine maneuvers. AB - BACKGROUND: Flying a helicopter requires greater coordination than flying a fixed wing aircraft, because the pilot is required to apply force simultaneously to three controls: the cyclic, collective, and pedals. There has been one study of pilot applied forces during helicopter flight, but this investigation did not examine muscle activity patterns. The aim of this study was to examine the muscle activation patterns and control forces of helicopter pilots during routine maneuvers. METHODS: Six pilots were tested in a UH-1 h Iroquois helicopter. The maneuvers involved hovering, winching, under-slung loads, a constant rate turn, and a high-speed valley turn. Variables recorded were pilot applied forces and electromyographic activity (EMG). Multivariate analysis of variance was used to test for differences between maneuvers. RESULTS: Significant differences between the maximum forces recorded from each control across all maneuvers were recorded (p < 0.05). The greatest pilot applied forces were recorded from the pedals. No muscles were activated more than 25% of a maximum voluntary contraction for any maneuver. The greatest magnitude of EMG activity was recorded from vastus lateralis during high-speed valley turns. There were significant differences between the EMG activity of left triceps, right triceps, and right deltoid for some maneuvers (p < 0.05). CONCLUSION: The control forces required to fly a helicopter during routine maneuvers are small. The levels of muscle activation associated with pilot applied forces are also low, but are similar to those reported during routine maneuvers in a fast-jet flight simulator. PMID- 10801000 TI - Head roll compensation in a visually coupled HMD: considerations for helicopter operations. AB - BACKGROUND: The helmet-mounted display (HMD) research program at the Flight Research Laboratory of the National Research Council of Canada examined the effects of HMD camera platform dynamics on pilot workload. Many currently fielded visually coupled HMD systems do not reproduce head movements in the roll axis which can lead to the presentation of visual information that is not consistent with vestibular and proprioceptive information. HYPOTHESIS: Our hypothesis was that this sensory conflict can induce motion sickness and increase pilot workload. METHODS: To examine this premise, three pilots flew a series of standardized maneuvers with or without roll compensation in the camera platform of a visually coupled HMD system. RESULTS: Increases in motion sickness symptoms and pilot workload were noted during complex, high-workload maneuvers when no roll compensation was present in the camera platform. During the most demanding maneuvers, the lack of roll compensation in the camera platform made it difficult for the evaluation pilot to control the helicopter. CONCLUSIONS: Roll compensation in visually coupled HMD systems reduces pilot workload and' motion sickness during critical flight periods where pilot workload may already be considerable. PMID- 10801001 TI - Cerebral blood flow velocity by transcranial Doppler during a vertical-rotating table simulation of the push-pull effect. AB - BACKGROUND: The push-pull effect (PPE) has been suspected of causing many aircraft accidents. The perfusion and then withdrawal of cerebral blood during the PPE may change the state of the cerebral blood vessel. HYPOTHESIS: During head-down tilt (HDT) cerebral vasoconstriction occurs in response to the elevated perfusion pressure to maintain cerebral blood flow, and during subsequent head-up tilt (HUT) the increased resistance of the cerebral blood vessel recovers slowly. METHODS: Ten healthy male non-pilots were exposed to the following protocol using a rotating-table to simulate the push-pull maneuver: HUT (+1 Gz) for 1 min followed by transition to HDT (-1 Gz) 10 s followed by transition to HUT (+1 Gz) 1 min. Cerebral blood flow velocity and pulsatility indices in the left middle cerebral artery were continually measured with a transcranial Doppler (TCD) instrument. RESULTS: Mean blood flow velocity (Vm) increased significantly by 10%, during the first 5 s of HDT, recovered to baseline during HDT 5 10 s, and remained unchanged during subsequent HUT. Systolic blood flow velocity (Vs) increased by 9% during HDT 5-10 s and 11% during HUT 0-5 s. Diastolic blood flow velocity (Vd) decreased by -9% during HDT 5-10 s, and -22% during HUT 0-5 s. Vs Vd increased by 26% during HDT 5 10 s, and 41%, during HUT 0-5 s. Pulsatile indices (PI) and resistance index (RI) increased by 26%) and 15% during HDT 5-10 s, and by 40% and 27% during HUT 0-5 s, respectively. Vs, Vs-Vd, PI, and RI remained at the higher level, and Vd remained at the lower level to HDT 15-20 s. CONCLUSIONS: The results indicate that cerebral vasoconstriction occurred to prevent brain over-perfusion during HDT. During HUT, the elevated resistance of the cerebral vessel remained at the higher level for about 20 s, and may have worsened the cerebral perfusion from exposure to +Gz. This may be one of the mechanisms of PPE. PMID- 10801002 TI - Post-roll effects on attitude perception: "the Gillingham Illusion". AB - BACKGROUND: Several aircraft each year are lost because of an unexplained collision with the ground. The attitude of most of these aircraft prior to impact was nose-low and with excessive bank, i.e., greater than 90 degrees . Prior to these accidents, each aircraft was noted as either changing heading or making an abrupt roll. HYPOTHESIS: Could there be some underlying tendency for the pilot to make unnoticed stick inputs after completing a roll from one bank angle to another? METHODS: Since ground-based flight simulators cannot create the true sensation of rolling an aircraft from one side to the other, the instrumented CALSPAN NT-33 aircraft was used for this study. Six pilots were given a series of three roll rates and two head positions while the aircraft automatically changed bank from 45 degrees of bank in one direction to 45 degrees of bank in the opposite direction. The subject's view of the external visual scene was restricted with a blue-amber vision restricting transparency combination. All attitude-indicating instruments were blanked, requiring the subjects to make stick inputs based on their vestibular (somatosensory) feedback. RESULTS: Subjects experienced a consistent tendency to increase bank angle after given control of the aircraft immediately following the roll maneuver, while thinking they were maintaining a constant bank angle. In some cases, the pilots rolled the aircraft completely inverted. CONCLUSION: When pilots rely on their perception of bank, following a roll, they will inadvertently increase their bank in the direction of the previous roll. PMID- 10801003 TI - Further support for the concept of a G-LOC syndrome: a survey of military high performance aviators. AB - METHODS: Some 329 military high-performance pilots were anonymously surveyed to determine the occurrence rates for a symptom complex of acceleration-induced neurologic manifestations. The premise for this symptom complex is the theory that acceleration-induced neurologic effects are not always an all-or-none phenomenon with G-LOC as the operational endpoint. RESULTS: A significant number of aircrew in selected types of aircraft reported symptoms such as euphoria, apathy, displacement, depersonalization, poor response to auditory stimuli, immediate memory difficulties, sensory abnormalities, motor abnormalities, confusion, and dream-like state without loss of consciousness. CONCLUSION: These findings may signal a need for alterations in G-awareness training. PMID- 10801004 TI - Pressurized sleeves and gloves for protection against acceleration-induced arm pain. AB - BACKGROUND: Acceleration (or G) induced arm pain may develop in centrifuge runs and in flight with low arm position and assisted pressure breathing during G (PBG) in combination with an extended coverage anti-G suit. To decrease this arm pain, pressurized sleeves and gloves were developed. METHODS: Eight subjects who earlier exhibited G-induced arm pain were tested on the centrifuge. The G exposures consisted of a gradual onset run up to a maximum of +9 G2, rapid onset runs to +3, +4, +5, +6, +7, +8, and +9 Gz and a simulated aerial combat maneuver (SACM) with peaks up to +9 Gz. On separate days, the subjects were tested without the sleeves and gloves, and with the sleeves and gloves pressurized to a maximum of 40, 60, or 80 mmHg at +9 Gz. The subjects reported their left and right arm pain on a subjective rating scale. RESULTS: G-induced arm pain, usually starting above +6 Gz, was often the reason for termination of the G-exposure without the pressurized sleeves and gloves. The pressurized sleeves and gloves significantly (p < 0.001) decreased arm pain, put no significant difference was found among the different pressures used. Heart rate was not different with and without the pressurized sleeves and gloves. CONCLUSIONS: The pressurized sleeves and gloves are an effective method to alleviate and sometimes eliminate G-induced arm pain. PMID- 10801005 TI - Mathematical models for predicting G-level tolerances. AB - The mathematical models developed in this article predict the following human G level tolerances: 1) rapid onset relaxed (ROR); 2) gradual onset relaxed (GOR); and, 3) straining-rapid onset. Included in the model are specific functions of: 1) anti-G suit; 2) positive pressure breathing (PBG); 3) baroreceptor reflex; 4) handgrip reflex; 5) anti-G straining maneuver (AGSM) increasing intrathoracic pressures (Pi); 6) leg elevation; and, 7) reclining seatback angles < or = 55 degrees. These functions are based on sound physiologic principles. Also discussed in the development of this model, but not included in the models, were: 1) isometric muscle contraction reflex; 2) Qigong (Q-G) maneuver; and, 3) straining GOR tolerances. The straining GOR tolerance profile was calculated to be a measure of G-duration tolerance and not G-level tolerance. A maximum P of 125 mm Hg from the AGSM was used in these models that could be augmented with PBG to 185 mm Hg. G-level tolerance predictions using this model were validated with published data. PMID- 10801006 TI - A mathematical model of cerebral perfusion subjected to Gz acceleration. AB - BACKGROUND: When the human body is exposed to a high gravitational load, the blood supply to the brain is reduced and loss of consciousness may occur. Our goal is to identify the principal mechanical causes of reduced blood supply to the brain during high +Gz. METHODS: We have developed a mathematical model to investigate the influence of Gz on the cerebral circulation. Blood flow is modeled using a one-dimensional flow approximation, in which the cross-sectional area of elastic vessels is determined as a non-linear function of the transmural (blood minus external) pressure. The intracranial vessels are subjected to cerebrospinal fluid pressure (PCSF) which is determined from the condition that the cranial volume is conserved. RESULTS: For a constant pressure difference of 100 mm Hg applied to the arterial and venous ends of the model, blood flow is diminished for +Gz. At approximately +5 G, the blood flow predicted by the model is insufficient to maintain normal functioning of the brain. PCSF is approximately equal to the blood pressure in the large intracranial veins for all values of Gz. Extracranial arteries and the intracranial vessels do not collapse, even when Gz is substantially higher than normal. However, the extracranial veins are collapsed even for moderate +Gz. CONCLUSIONS: Even if cardiac output is maintained at normal levels, cerebral perfusion will fall because of the increasing resistance of the cerebral flow circuit. This increase is largely due to the collapse of the extracranial veins, which begins at moderate Gz and becomes dominant at a Gz of approximately 4.5. PMID- 10801007 TI - The experience of nausea during sustained hyper-gravity flight with negligible angular velocity. AB - BACKGROUND: The purpose of this study was to investigate nausea and vomiting during hyper-gravity flight at a slow rate of turn. During head-movements under these conditions, the semi-circular canals of the vestibular system function normally whereas the otoliths experience a G-excess effect, displacing further and moving faster than in a 1 G field. HYPOTHESIS: Nausea and vomiting are greater during hyper-gravity flight compared with historical data collected in a 1 G field at a similar rate of turn. METHODS: There were 27 subjects who were exposed to 1.8 G (hyper-G) during a slow rate turn on three NASA KC-135 flights. Subjects participated in one of three experimental periods, each period consisting of two 5.5-min hyper-G runs: 1) rest, with only incidental head movements; 2) active roll and pitch head movements; or 3) passive roll head movements. Subjective symptom data were collected pre-flight, following each experimental period and post-flight using a standardized nausea questionnaire. Electrogastrograms were obtained from eight subjects. RESULTS: Seven subjects (26%) vomited during the flight. Nausea initially increased but was then stable throughout the flight. Nausea appeared highest during active head movements. In subjects who vomited, greater gastrointestinal distress and somatic distress were reported. CONCLUSIONS: The levels of nausea and vomiting observed during hyper-G cannot be explained by Coriolis cross coupling and are likely due to the G-excess effect on the otoliths. The nausea profiles observed in individuals who vomit during hyper-G appear similar to those previously observed during nausea produced by an optokinetic drum stimulus in a 1-G field. PMID- 10801008 TI - Blood pressure and sympathetic activity in normotensive aviators during short haul fixed-wings flights. AB - BACKGROUND: In order to evaluate the impact of in-flight stress on BP and sympathetic activity during a short fixed-wing flight, we have studied 18 healthy and normotensive commercial captain aviators, aged 37.5 +/- 4.9 yr. METHODS: The protocol consisted of a 120-min flight period, divided into segments of pre flight, take off, mid-cruise, approach and landing, and a 120-min control period. In both flight and control periods, all subjects underwent BP monitoring, heart rate recording and urine collection for catecholamines. RESULTS: Systolic and diastolic BP (SBP/DBP) were higher during the flight, as a whole, when compared with the control period (134 +/- 11 vs. 121 +/- 8 mmHg, p < 0.05 and 84 +/- 8 vs. 76 +/- 9 mmHg, p < 0.05, respectively). During the flight period, SBP increased in the pre-flight, take-off, approach and landing segments, whereas DBP increased in the take-off, mid-cruise, approach and landing segments. Heart rate did not change in any flight segment. Urinary catecholamines increased during the flight period in comparison to control period 0.20 (0.10 1.8) mg x mg-1 creatinine vs. 0.10 (0.10-1.0) mg x mg-1 creatinine; p < 0.05). CONCLUSION: We conclude that inflight stress increases BP of the normotensive aviators by sympathetic activation during short-haul flights. PMID- 10801009 TI - Release from masking in virtual auditory space during sustained positive acceleration. AB - BACKGROUND: The use of a three-dimensional (3-D) auditory display to significantly lower subject detection level while maintaining comprehension under sustained positive G-stress was explored in this study. METHODS: Auditory threshold levels were measured for detecting a band limited pulsed signal in the presence of a broadband diotic masker at both + 1 Gz (rest position) and under sustained +3 Gz. The pulsed signal was presented diotically and was spatialized at one of four static azimuth positions on the horizontal plane. RESULTS: Results showed that auditory thresholds were not significantly affected by sustained +3 Gz stress. Compared with a diotic presentation, subjects reached an average of 6.8 dB lower auditory threshold at +1 Gz and under sustained +3 Gz when the pulsed signal was spatialized at a static position of 90 degrees azimuth on the horizontal plane. CONCLUSION: The implication of these results and suggestions for further research are discussed. PMID- 10801010 TI - Hereditary hemochromatosis among U.S. Army aviators. AB - From 1988 to 1995, there were five reported cases of hemochromatosis among U.S. Army aviators. Three of the five are presented and discussed. The cases of hereditary hemochromatosis were discovered during unrelated work-ups or from investigation of a positive family history, and not by routine flying duty medical examinations. Recent studies show a prevalence of 5 per 1000 in the general population. This study shows the incidence among Army aviators is 0.296 per 1,000 aviator-years of observation. It is possible that there are cases presently undiagnosed in the Army aviation community. Without screening measures in place, the Army Aviation Branch has greater difficulty diagnosing and treating hereditary hemochromatosis. Heightened awareness and a high clinical index of suspicion are necessary to identify affected patients. Early detection and treatment is essential to prevent long-term end organ damage from iron deposition. PMID- 10801011 TI - Airlines emergency medical kits. PMID- 10801012 TI - Raynaud's phenomenon and flying. PMID- 10801013 TI - Ejection associated injuries. PMID- 10801014 TI - Airline use of automatic external defibrillator: shocking developments. PMID- 10801015 TI - A tourist in space. PMID- 10801016 TI - Searching Entrez PubMed and uncover on the internet. PMID- 10801017 TI - Gifts from surgical research. Contributions to patients and to surgeons (2) PMID- 10801018 TI - Stage 0 to stage III breast cancer in young women. AB - BACKGROUND: Breast cancer survival is improving because mammography is leading to diagnosis at earlier stages of the disease. Because young women with breast cancer rarely undergo mammography before diagnosis, outcomes for breast cancer in young women may not be improving. In addition to advanced stage, young age at diagnosis is associated with biologically more aggressive cancers with higher rates of local and distant recurrence. STUDY DESIGN: Risk factors, clinical presentations, pathologic findings, tumor characteristics, extent of disease, treatment, and outcomes for 101 women under age 36 treated for breast cancer between 1989 and 1997 were compared with 631 patients 36 years and older treated by us during the same interval. Stage IV patients were excluded. RESULTS: Patients younger than 36 years were more likely to present with a palpable mass (87% versus 55%, p < 0.001) and were less likely to undergo spot localization breast biopsy for mammographic findings (40% versus 6%, p < 0.001). Patients younger than 36 years had larger tumors (median 2.0 cm versus 1.5 cm, p < 0.001), more nodal involvement (50% versus 37%, p = 0.022), more nodes involved (median 1.0 versus 0, p = 0.010), and were more likely to be diagnosed with stage II or III cancer (60% versus 43%, overall p < 0.001). Young patients' cancers were more poorly differentiated (80% versus 44%, overall p < 0.001), estrogen receptor negative (52% versus 31%, p < 0.001), aneuploid (70% versus 49%, p = 0.013), and had higher S-phase fractions (59% versus 29%, p = 0.001). Patients less than 36 years were treated more often with mastectomy (59% versus 22%, p < 0.001) and adjuvant chemotherapy (80% versus 54%, p < 0.001) and less often with tamoxifen (36% versus 58%, p = 0.001). Cumulative 5-year local and distant disease-free survival were significantly worse for patients younger than 36 years (p = 0.011 and p = 0.044, respectively). The higher rate of local recurrence in patients less than 36 years was from an excess number of local recurrences in patients treated with breast conservation. After consideration for nodal involvement, chemotherapy, and tamoxifen using the Cox proportional hazards model, no other variable, including age, was significantly related to local disease-free outcomes. After consideration for tumor size and nodal involvement, no other variable was significantly related to distant disease failure rates. CONCLUSIONS: Patients diagnosed with breast cancer before age 36 differ from older patients in numerous respects. They present more often with a palpable mass rather than a mammographic finding and their cancers are more advanced with features that are more aggressive. Despite aggressive treatment, most commonly with mastectomy and chemotherapy, local and distant failure rates are higher than for patients 36 and older. The higher rate of local recurrence in patients less than 36 years reflects an excess number of local recurrences in patients treated with breast conservation. PMID- 10801019 TI - Incidence of chromosomes 1 and 17 aneusomy in breast cancer and adjacent tissue: an interphase cytogenetic study. AB - BACKGROUND: Characterization of the biopathologic events underlying the early steps of breast carcinogenesis may have a dramatic impact on reducing breast cancer mortality. Genes involved in breast tumorigenesis are localized on chromosomes 1 and 17, and numeric aberrations of these chromosomes have been correlated with breast cancer tumorigenesis and progression. According to the field cancerization hypothesis, specific chromosome aberrations may be present in breast cancer and in normal-appearing adjacent tissue. The latter changes reflect the genomic damage that follows longterm carcinogenic exposure and precede the morphologically detectable neoplastic transformation. We hypothesize that detection of these aberrations in benign breast epithelium may provide a tool for molecular risk assessment. STUDY DESIGN: Using fluorescence in situ hybridization with centromere-specific probes, we determined the status of chromosomes 1 and 17 in fresh imprints of 28 samples of primary tumors and 54 samples of their surrounding uninvolved parenchyma taken from patients undergoing operations for breast carcinoma. Ten contralateral breast biopsy specimens collected from patients with previous breast carcinoma were also evaluated as a surrogate of a high-risk group to rule out the hypothesis that chromosomal aneusomy in tumor adjacent tissue could be related to a paracrine effect of the primary tumor. Ten samples of benign breast tissue taken from patients at low risk were used as controls to define tolerance limits for aneusomy definition. RESULTS: Using threshold values of 40% of signal loss and 13% of signal gain to define chromosome aneusomy (ie, mean + 3 SDs of the control group signals), we found the following: 1) almost all primary breast tumors were aneusomic for chromosomes 1 and 17; 2) primary breast tumor and adjacent uninvolved parenchyma shared the same pattern of chromosomes 1 and 17 aneusomy in 66.7% of patients; and 3) chromosomes 1 and 17 aneusomies in contralateral benign breast samples from high risk patients were not different from those in primary breast tumor or adjacent tissue samples. CONCLUSIONS: These results suggest that chromosomes 1 and 17 aneusomy may represent an intermediate biomarker of breast tumorigenesis potentially useful to detect patients at high risk of breast carcinoma who may benefit from preventive interventions. PMID- 10801020 TI - The utility of sestamibi scanning in the operative management of patients with primary hyperparathyroidism. AB - BACKGROUND: The role of imaging studies before parathyroidectomy has been extensively debated and recent advances in unilateral parathyroidectomy intensify this controversy. The purpose of this study was to review the parathyroidectomy experience of a single surgeon, looking at the role of sestamibi scans and a standard postoperative care regimen. STUDY DESIGN: Retrospective review of office and hospital charts was completed on 90 patients with primary hyperparathyroidism who underwent parathyroidectomy from 1991 to 1998. Patient workup and outcomes were noted, as were results of preoperative imaging. True-positive scans visualized an abnormality ipsilateral to the adenoma found at operation. Statistics were performed using nonparametric testing and Student's t-test. RESULTS: There were 21 male and 69 female patients, with an average age of 54 years (range 29 to 81). There were zero mortalities, three morbidities (3.3%), and three patients who had persistent hypercalcemia, yielding a 96.7% success rate. Sixty-seven patients underwent preoperative sestamibi scanning, with a sensitivity of 74% and positive predictive value of 89%. Operative time in imaged patients averaged 103 +/- 49.9 minutes versus 121.5 +/- 85.9 minutes for patients without sestamibi scans. Operating time differences were not statistically significant and a preoperative sestamibi scan did not affect the success of parathyroidectomy. Discharge on postoperative day 1 was accomplished in 80% of patients and 13% were discharged the next day. There was no morbidity from hypocalcemia. CONCLUSIONS: A preoperative sestamibi scan does not improve efficacy or decrease operating time for primary hyperparathyroidism when bilateral neck exploration is performed. A postoperative care protocol including oral calcium and vitamin D supplementation allows the majority of patients to be discharged on postoperative day 1 with excellent results. PMID- 10801021 TI - Hyperparathyroidism after thyroid surgery and autotransplantation of histologically normal parathyroid glands. AB - BACKGROUND: Parathyroid autotransplantation is a well-established method to prevent hypoparathyroidism during parathyroid and thyroid operations. The reported success rate of parathyroid autotransplantation ranges from 75% to 100%. Recurrent hyperparathyroidism may develop after parathyroid autotransplantation, especially after the transplantation of hyperplastic or adenomatous parathyroid tissue. Hyperparathyroidism recurs most frequently after subtotal parathyroidectomy or total parathyroidectomy and autotransplantation, in patients with renal failure and secondary hyperparathyroidism, and in patients with familial primary hyperparathyroidism or MEN I or MEN II syndrome. We report three patients who experienced primary hyperparathyroidism after autotransplantation of normal parathyroid tissue during thyroid operations (two patients) or after a long period of hypoparathyroidism. STUDY DESIGN: We reviewed our records from 1983 to May 1998 and identified three patients in whom hyperparathyroidism developed after thyroid operations. RESULTS: One patient had a thyroidectomy with left modified radical neck dissection for papillary thyroid cancer, followed by radioiodine ablative therapy. Two patients had thyroid operations for benign thyroid disease. One of these patients had a history of radiation exposure for acne, and in the other one secondary hyperparathyroidism arose 6 years after a thyroidectomy for hyperthyroidism. CONCLUSIONS: Our study documents that hyperparathyroidism may develop after autotransplantation of histologically normal parathyroid tissue and after a period of hypoparathyroidism after thyroid operations. For this reason, it is important to mark the site of the parathyroid transplantation. PMID- 10801022 TI - Laparoscopic repair of large type III hiatal hernia: objective followup reveals high recurrence rate. AB - BACKGROUND: Recent studies based on symptomatic outcomes analyses have shown that laparoscopic repair of large type III hiatal hernias is safe, successful, and equivalent to open repair. These outcomes analyses were based on a relatively short followup period and lack objective confirmation that the hernia has not recurred. The aim of this study was to compare the outcomes of laparoscopic and open repair of large type III hiatal hernia using both symptomatic evaluation and barium study to assess the integrity of the repair. STUDY DESIGN: Fifty-four patients underwent repair of a large type III hiatal hernia between 1985 and 1998. The surgical approach was laparotomy in 13, thoracotomy in 14, and laparoscopy in 27. An antireflux procedure was included in all patients. Symptomatic outcomes were assessed using a structured questionnaire at a median of 24 months and was complete in 51 of 54 patients (94%). A single radiologist, without knowledge of the operative procedure, assessed the integrity of the repair using video esophagram. Videos were performed at a median of 27 months (35 months open and 17 laparoscopic) and were completed in 41 of 54 patients (75%). RESULTS: Symptomatic outcomes were similar in both groups with excellent or good outcomes in 76% of the patients after laparoscopic repair and 88% after an open repair. Reherniation was present in 12 patients and was asymptomatic in 7. A recurrent hernia was present in 12 of the 41 patients (29%) who returned for a followup video esophagram. Forty-two percent (9 of 21) of the laparoscopic group had a recurrent hernia compared with 15% (3 of 20) of the open group (p < 0.001 log-rank value on recurrence-free followup). CONCLUSIONS: Laparoscopic repair of type III hiatal hernias is associated with a disturbingly high (42%) prevalence of recurrent hernia. More than half such recurrences have few, if any, symptoms. PMID- 10801023 TI - Esophageal cancer: results of an American College of Surgeons Patient Care Evaluation Study. AB - BACKGROUND: The last two decades have seen changes in the prevalence, histologic type, and management algorithms for patients with esophageal cancer. The purpose of this study was to evaluate the presentation, stage distribution, and treatment of patients with esophageal cancer using the National Cancer Database of the American College of Surgeons. STUDY DESIGN: Consecutively accessed patients (n = 5,044) with esophageal cancer from 828 hospitals during 1994 were evaluated in 1997 for case mix, diagnostic tests, and treatment modalities. RESULTS: The mean age of patients was 67.3 years with a male to female ratio of 3:1; non-Hispanic Caucasians made up most patients. Only 16.6% reported no tobacco use. Dysphagia (74%), weight loss (57.3%), gastrointestinal reflux (20.5%), odynophagia (16.6%), and dyspnea (12.1%) were the most common symptoms. Approximately 50% of patients had the tumor in the lower third of the esophagus. Of all patients, 51.6% had squamous cell histology and 41.9% had adenocarcinoma. Barrett's esophagus occurred in 777 patients, or 39% of those with adenocarcinoma. Of those patients that underwent surgery initially, pathology revealed stage I (13.3%), II (34.7%), III (35.7%), and IV (12.3%) disease. For patients with various stages of squamous cell cancer, radiation therapy plus chemotherapy were the most common treatment modalities (39.5%) compared with surgery plus adjuvant therapy (13.2%). For patients with adenocarcinoma, surgery plus adjuvant therapy were the most common treatment methods. Disease-specific overall survival at 1 year was 43%, ranging from 70% to 18% from stages I to IV. CONCLUSIONS: Cancer of the esophagus shows an increasing occurrence of adenocarcinoma in the lower third of the esophagus and is frequently associated with Barrett's esophagus. Choice of treatment was influenced by tumor histology and tumor site. Multimodality (neoadjuvant) therapy was the most common treatment method for patients with esophageal adenocarcinoma. The use of multimodality treatment did not appear to increase postoperative morbidity. PMID- 10801024 TI - Perioperative safety and prognosis in hepatocellular carcinoma patients with impaired liver function. AB - BACKGROUND: The benefits of liver resection for hepatocellular carcinoma (HCC) patients with concomitant impaired liver function were often considered questionable because of poor postoperative prognosis. This study will clarify whether an acceptable operative risk exists and whether limited resection will compromise the outcomes of these patients. STUDY DESIGN: Between July 1991 and December 1996, a total of 168 patients with HCC who underwent hepatectomies were enrolled and divided into normal (group A) and impaired (group B) liver function groups according to the value of indocyanine green retention rate at 15 minutes. Clinical features, surgical related features, pathologic features, and disease free and overall survivals were compared between the groups. RESULTS: Operative morbidity and mortality in group A were 27.3% and 1.6%, and in group B were 40.0% and 2.5%, respectively (p = 0.129 and 0.506). Disease-free survival and overall survival at 5 years in group A were 43.2% and 59.6%, respectively, and in group B they were 30.6% and 56.8%, respectively (p = 0.607 and 0.378). CONCLUSIONS: Limited liver resection is safe and provides favorable prognosis in HCC patients with concomitant impaired liver function. PMID- 10801025 TI - Resection or transplantation for hepatocellular carcinoma in cirrhotic patients: outcomes based on indicated treatment strategy. AB - BACKGROUND: Surgical resection has been the treatment of choice for hepatocellular carcinoma (HCC), but the resection rate remains low in cirrhotic patients and recurrence is common. Unfavorable results compared with benign disease and the shortage of organ donors have led to a restricted indication for orthotopic liver transplantation (OLT) for HCC. STUDY DESIGN: The aim of this study was to analyze the results of our surgical approach to HCC in patients with cirrhosis. The first treatment strategy indicated in these patients was OLT. From January 1990 to May 1999, 85 patients underwent OLT and the remaining 35 had surgical resection. RESULTS: One-, 3-, and 5-year survival rates were 84%, 74%, and 60% versus 83%, 57%, and 51%, respectively, in the OLT and resection groups (p = 0.34). Hepatic tumor recurrence was much less frequent in the OLT group than in the resection group. The 1-, 3-, and 5-year disease-free survival rates were 83%, 72%, and 60% versus 70%, 44%, and 31%, respectively (p = 0.027). In the multivariate Cox regression analysis, macroscopic vascular invasion was the only factor independently associated with death or recurrence after OLT (p = 0.006). After partial liver resection, the tumors significantly associated with mortality and recurrence in the multivariate analysis were solitary or multiple tumors greater than 2cm with microscopic vascular invasion (pathologic pT3) (p = 0.01). CONCLUSIONS: Our results confirm that in cirrhotic patients, OLT may provide better outcomes than liver resection in carefully selected HCC and that longterm survival is similar to the results of OLT in cirrhotic patients without tumors. PMID- 10801026 TI - Refractory abdominal-cutaneous fistulas or leaks: percutaneous management with a collagen plug. AB - BACKGROUND: We report the results of abdominal-cutaneous fistula tract occlusion with a collagen plug in a series of patients with fistulas or leaks refractory to conservative therapy. STUDY DESIGN: Seven patients were found to have persistent fistula or leak after percutaneous drainage of abdominal pelvic fluid collections. All patients but one were refractory to surgical or percutaneous drainage. Under fluoroscopic guidance, modified Vasoseal (Datascope Inc, Montvale, NJ) collagen plugs were deployed into the fistulas using catheter directed techniques. The plugs were split longitudinally to fit into an 8F or 9F peel-away sheath, placed into the fistula, and deployed. Results were tabulated and patients were followed up. RESULTS: Six of seven patients undergoing fluoroscopically guided, catheter-directed tract occlusion had resolution of the fistula, with no evidence of fistula or abscess recurrence from 30 to 180 days after closure. There were no procedural complications. The technique was unsuccessful in dosing a gastrocutaneous fistula after removal of a large-bore gastrostomy tube; this failure was believed to be secondary to the short length and large caliber of the tract in a patient with hypercortisolemia. CONCLUSIONS: Closure of abdominal-cutaneous fistula tracts by occlusion with a modified Vasoseal collagen plug shows promise in the management of fistulas refractory to catheter drainage. PMID- 10801027 TI - The army at Pearl Harbor. PMID- 10801028 TI - The acute and chronic management of spinal cord injury. PMID- 10801029 TI - The role of genetic screening and prophylactic surgery in surgical oncology. PMID- 10801030 TI - A rare cause of complete small-bowel obstruction. PMID- 10801031 TI - Magnetic resonance cholangiopancreatographic diagnosis of Mirizzi's syndrome. PMID- 10801032 TI - Hilar plate detachment and extraglissonian extrahepatic anterior approach to the right portal pedicle for right liver resections. PMID- 10801033 TI - Living-related transplantation of left liver plus caudate lobe. PMID- 10801034 TI - The dictated operative note: important but is it being taught? PMID- 10801035 TI - Biotypes of oral Candida albicans isolated from AIDS patients and HIV-free subjects in Thailand. AB - This study was conducted to examine biotypes and antifungal susceptibility patterns of oral Candida albicans isolated from HIV-infected patients, HIV-free patients with candidiasis and healthy subjects. All isolates were biotyped using a typing system based on enzyme profiles, carbohydrate assimilation patterns and boric acid resistance. Thirty-eight biotypes were found amongst 218 oral C. albicans isolates. The major biotype found was A1S, which accounted for 32.6% of all isolates, and this biotype was the most common in all groups. There was a greater variety of biotypes of C. albicans in the HIV-infected group than in the other groups; however, there was no statistically significant difference between the groups. The minimum inhibitory concentrations (MICs) of a total of 118 isolates were determined for amphotericin B and for ketoconazole using the National Committee for Clinical Laboratory Standards (NCCLS) broth macrodilution method and the E-test. When the antifungal susceptibility patterns among the groups were compared, a statistically significant difference was found only with amphotericin B. The median MIC of amphotericin B in the HIV-infected group was higher than in the healthy group (P=0.013, NCCLS method; P=0.002, E-test). However, this difference in sensitivity was not restricted to any sub-type investigated. Our results showed that the biotype patterns of C. albicans isolates that colonize HIV-infected patients are similar to those of HIV-free subjects, and there is no relationship between antifungal susceptibility patterns and the biotypes. PMID- 10801036 TI - Effects of date extract on adhesion of Candida species to human buccal epithelial cells in vitro. AB - The adherence of three Candida species to human buccal epithelial cells (BEC) following treatment with different concentrations of date extract was investigated in vitro, as well as the effect of a mouth rinse with date extract on the adhesion of yeast to BEC. Adhesion of C. albicans, C. tropicalis and C. kefyr to BEC was significantly reduced after both short- and long-term periods of yeast exposure to various concentrations of date extract (reduction between 25% and 52% of the control value). A similar inhibition of adherence was observed upon pre-incubation of BEC with date extract. There was a significant reduction (P<0.001) in the adherence of yeast to BEC collected immediately or 5-20 min after an oral rinse with 10% date extract. No statistically significant difference was observed in the adhesion of BEC collected 30 min after an oral rinse with date extract and control BEC. In addition, pre-treatment of either Candida or BEC, or both, with date extract resulted in reduced adherence, the magnitude of which was largest when both types of cells were pre-treated. Date extract also inhibited germ-tube formation of C. albicans (56-85% inhibition), which might contribute to the effects on adherence. PMID- 10801037 TI - Sub-therapeutic exposure to polyene antimycotics elicits a post-antifungal effect (PAFE) and depresses the cell surface hydrophobicity of oral Candida albicans isolates. AB - Post-antifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of fungi to antimycotics and subsequent removal of the drug. The fungal pathogen Candida albicans is the major aetiologic agent of oral candidosis, and the cell surface hydrophobicity (CSH) of this yeast is considered a critical factor contributing to its colonisation potential. As the concentration of topically prescribed antifungals reach sub-therapeutic levels at dosage intervals, the study of the polyene-induced PAFE and its impact on the CSH of oral C. albicans should be of clinical relevance. Hence the aims of this investigation were to measure the PAFE and CSH of 12 isolates of C. albicans following limited exposure (1 h) to nystatin and amphotericin B and also to investigate the ultrastructural features of yeast cells following such antifungal exposure. The yeasts were exposed to sub-lethal concentrations of nystatin (x2 MIC) and amphotericin B (x2 MIC) for a period of 1 h. Following subsequent removal of the drug, the PAFE and the CSH of the isolates were assessed by a turbidometric measurement of growth and a biphasic aqueous-hydrocarbon assay, respectively. The mean duration of PAFE of nystatin and amphotericin B were 5.99 (+/-0.49) h and 8.73 (+/-0.93) h, respectively, while the reduction in CSH following exposure to these drugs were 17.32% (P<0.05 for 83% of the isolates) and 14.26% (P<0.05 for 66% of the isolates), respectively. On scanning electron microscopy the exposed cells were seen to undergo collapse of the internal cell membrane, leaving an intact cell wall, while a proportion of cells were deflated. Some cells showed intense puckering of the cell wall, resulting in a mulberry appearance. Taken together, these data elucidate additional mechanisms by which polyene antimycotics may operate in vivo to suppress candidal pathogenicity. PMID- 10801038 TI - Predictors of tobacco and alcohol consumption and their relevance to oral cancer control amongst people from minority ethnic communities in the South Thames health region, England. AB - The purpose of this study was to examine the determinants of the health behaviour of ethnic groups in relation to alcohol and tobacco use. A cross-sectional questionnaire survey was carried out using network sampling amongst community groups in the South Thames region of the United Kingdom. Self-classified ethnic groups were identified: Black-African; Black-Caribbean; Indian; Pakistani; Bangladeshi and Chinese/Vietnamese. A total of 1113 people were recruited in the study. In all of the ethnic groups, men were more likely than women to smoke tobacco. Chewing of pan and tobacco was common in the South Asian communities and alcohol consumption was high among the Black-Caribbean group. Those factors were predicted by education, employment, gender and being born in the UK. It is important to examine the determinants of such risk behaviours in order to aid appropriate targeting of health promotion interventions, particularly those related to cancer control. PMID- 10801039 TI - Fluid phase endocytosis in oral epithelia: variation with site and effect of cancer. AB - The structure of the oral mucosa has been extensively studied but its cell physiology has been less well characterised. This study aimed to show the range in variation in fluid phase endocytic capability in biopsies from different oral sites. Oral epithelial cells were obtained from both biopsies and single-cell suspensions obtained by brushing the oral cavity. Biopsies in organ culture and single cells in suspension were incubated with fluorescent microspheres of 0.02, 0.1 or 1.0 microm diameter. Endocytosis of fluorescent microspheres was quantitated by flow cytometry and visualised by confocal microscopy. Epithelial cells from all oral sites that were sampled internalised 0.02 microm and 0.1 microm but not 1.0 microm microspheres, with no significant differences observed between oral regions. Single cells from non-cancer patients endocytosed significantly more 0.02 microm microspheres than cells removed from patients with oral cancer. This model may be used to study integrated oral cell function both in health and disease. PMID- 10801040 TI - The relationship of genetic aberrations detected by comparative genomic hybridization to DNA ploidy and tumor size in human oral squamous cell carcinomas. AB - We have examined genetic alterations in 11 surgically removed oral squamous cell carcinomas (OSCCs) using comparative genomic hybridization (CGH) and laser scanning cytometry (LSC), which allow quantitative analysis of chromosomal abnormalities. CGH analysis revealed gains and/or losses of DNA sequence copy number in all tumors. Gains in DNA sequence copy number were detected frequently for chromosome arms 3q25-28 (6/11), 5p (6/11) and 8q (5/11), and losses in chromosome arms 18q (4/11), 19q (4/11), 17p (3/11), and 19p (3/11). Amplification of 5p was observed in two tumors. LSC detected DNA aneuploidy with DNA indices ranging from 1.30 to 1.82 in 6 of 11 tumors. The number of chromosomal aberrations was higher in DNA aneuploid tumors than in diploid tumors (8.17 vs 3.60/tumor, P<0.05). Furthermore, the average number of chromosomal aberrations was significantly higher in stage T2 tumors and larger tumors than in stage T1 tumors (7.71 vs 3.25/tumor, P<0.05). Our results suggest that DNA aneuploidy and large tumor size reflect an underlying chromosomal instability. PMID- 10801041 TI - Endogenous TGF-beta1 inhibits the growth and metastatic dissemination of rat oral carcinoma cell lines but enhances local bone resorption. AB - This study examined the effect of stable transfection of latent transforming growth factor-beta1 (TGF-beta1) cDNA into a predominantly polygonal, 4 nitroquinoline N-oxide (4NQO)-induced rat oral keratinocyte cell line. Seven polygonal and five spindle clonal populations were isolated that overexpressed TGF-beta1 protein by approximately two- to four-fold compared to vector-only transfected controls. Neutralisation experiments indicated that the majority of protein was in the latent form. There was no change in the proportion of polygonal and spindle cells in vitro after transfection with TGF-beta1 cDNA. Polygonal and spindle cells that overexpressed TGF-beta1 produced similar amounts of protein and grew more slowly in vitro than controls. The parent cell line and all transfected cells were growth inhibited (60-75%) by exogenous TGF-beta1. Orthotopic transplantation of the parent and the vector-only control cell lines resulted in primary tumours in the floor of the mouth in almost 100% (20/21) of athymic mice, with no evidence of bone resorption at the site of the primary tumour and pulmonary metastatic tumour deposits in some 40% (7/20) of these animals. The polygonal and spindle cells that overexpressed TGF-beta1 behaved similarly following orthotopic transplantation. A 96% (23/24) primary tumour take was evident following transplantation of cells that overexpressed TGF-beta1, with a significantly (P<0.02) higher number of animals showing bone resorption at the site of the primary tumour (35%; 8/23) compared to controls. By contrast, there was a significant (P<0.03) decrease in the number of animals with pulmonary metastases (4%; 1/23) following transplantation of TGF-beta1 overexpressing cells compared to controls. Overexpression of TGF-beta1 did not alter tumour cell differentiation in vivo. The results demonstrate that endogenous TGF-beta1 functions as a tumour suppressor in the rat-4NQO model of oral carcinogenesis without altering tumour cell morphology or differentiation but can also act to promote local bone resorption. PMID- 10801042 TI - Ivan Petrovich Pavlov. PMID- 10801043 TI - Pulmonary aspiration complicating esophagogastric anastomoses in rats. PMID- 10801044 TI - Modified technique for heterotopic rat heart transplantation under cardioplegic arrest. AB - The development of microsurgical techniques offers a valuable opportunity to use small animals for experimental studies of vascularized organ transplants. Availability of inbred strains, natural resistance to infection, and economy make the rat an ideal animal model to investigate the effects of heart transplantation. The recent high interest and substantial laboratory activity with regard to posttransplantory immunological tissue reactions and apoptotic tissue processes led us to optimize transplantation technique by improving myocardial protection during ischemia and thereby minimizing adverse effects of the transplantation procedure itself. Thus the present report details the technique of heterotopic heart transplantation in rats using cardioplegic arrest. PMID- 10801045 TI - Hemoglobin solutions as blood substitutes. PMID- 10801046 TI - Biochemical alterations in rat Thiry-Vella fistulas. AB - In order to obtain baseline information on the secretory function of normal rat bowel for our work on intestinal graft ischemia, we studied several biochemical parameters in rat Thiry-Vella fistulas (TVF). TVFs were created in 200-g male Lewis rats (n = 11) using the 25-cm segment of jejunum normally used as a graft in our intestinal transplant model. The stomas were matured primarily and the animals were allowed to recover. The TVFs were flushed at 0, 6, and 24 h and then daily for up to 21 days with 12 mL normal saline solution. The effluent was collected and analyzed for total protein (TP), secretory phospholipase A2 (sPLA2), intestinal fatty acid binding protein (I-FABP), lactate dehydrogenase (LDH), and N-acetylglucosamine (NAGA). TP content was 0.12 +/- 0.01 mg/mL up to 48 h, then gradually increased and stabilized at 0.39 +/- 0.05 mg/mL at day 21. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), one major protein band was identified in the low-molecular-mass range (15 kD), consistent with I-FABP and sPLA2. Secretory PLA2 levels decreased over the first 4 days to a low of 115 +/- 24.8% hydrolysis/min/fraction, then gradually rose to a plateau at approximately 529.76 +/- 88.36% hydrolysis/min/fraction by day 18. I FABP levels rose rapidly from 0 ng/mL at 2 h to 900 +/- 250.0 ng/mL at 6 h and approximately 3000 +/- 304.9 ng/mL by day 14. LDH levels at 2 h and 48 h did not differ, with 0.03 +/- 0.004 and 0.03 +/- 0.005 optical density units (OD)/min/mL, respectively. NAGA levels were 0.07 +/- 0.05 OD/h/mL at 2 h and rose to 0.14 +/- 0.04 OD/h/mL at 48 h. These data suggest that after an early equilibration period, biochemical secretion into the lumen of normal rat bowel reaches a state of equilibrium, and therefore appears to reflect the baseline biochemical status of the bowel. Some of these levels are not negligible as one would expect in "normal" bowel. This information should prove extremely helpful as a baseline study of abnormal conditions of the intestine, such as ischemia or rejection. PMID- 10801047 TI - Effect of magnetic stimulation on the electromechanical activity of the rectum: an experimental study. AB - We recently studied the effect of sacral magnetic stimulation (MS) on the rectal mechanical activity. The current communication investigates its effect on the rectal electric activity. Twelve mongrel dogs (weight 15.2 +/- 3.2 kg, 7 male, 5 female) were anesthetized, the abdomen was opened, and three electrodes were sutured to the rectal serosa. The rectal pressure was recorded by a manometric catheter. The effect of sacral MS on the rectal electric activity and pressure was recorded before and after anesthetization of the sacral skin. Stimulation parameters were set at 70% intensity, 20 Hz frequency, and 1 to 5 s burst activity. Sacral MS of the empty and full rectum effected a significant increase of the rectal electric activity and pressure as well as a decrease of the rectal neck (anal canal) pressure. MS of the anesthetized sacral skin produced no significant changes in the rectal electromechanical activity or the anal canal pressure. MS of the saline-injected sacral skin produced rectal and anal changes similar to those without saline injection. In conclusion, MS presumably produces its effect on the rectum by increasing the rectal electric activity. It might provide a more convenient method for rectal evacuation than electrostimulation through implanted electrodes. The procedure is simple, easy, and noninvasive. PMID- 10801048 TI - A sheep model for thoracic aortic surgery in the presence of systemic coagulopathy. AB - Surgical repair of aneurysms, traumatic injuries, or congenital anomalies of the thoracic aorta are associated with high morbidity and mortality mainly as a result of excessive and uncontrollable hemorrhage from diffuse coagulopathy. We developed a model in sheep that simulates this coagulopathic state for experimentation with thoracic aorta surgery. This experimental animal model involves administering a 600-mg aspirin suppository once a day for the 2 days preceding surgery and a final dose on-call to surgery. Prior to cross-clamping the aorta, an intravenous (i.v.) bolus of heparin (400 IU/kg) was administered. Thirty minutes later, the i.v. heparin bolus was repeated. Pre- and intraoperative activated clotting time was 101 +/- 10 s and >1500 s (p < .0001); prothrombin time, 21 +/- 1 s and >100 s (p < .0001); and activated partial thromboplastin time, 20 +/- 1 s and >50 s (p < .0001), respectively. We utilized a partial cross-clamp-and-sew technique to anastomose a woven, gelatin impregnated, 16-mm tube graft end-to-side to the descending thoracic aorta. Mean total blood loss was 1367 +/- 282 mL, which included mean blood loss from time of release of aortic cross-clamp to close (422 +/- 135 mL) and mean total blood output from chest tube drain (945 +/- 203 mL). The mean time to achieve hemostasis at suture lines after aortic cross-clamp release was 15.5 +/- 6.6 min. In conclusion, a sheep model with induced coagulation defects was successfully developed and reproducible for experimentation involving thoracic aortic surgery. PMID- 10801049 TI - Competitive NMDA receptor antagonists and spinal-cord ischemia. AB - Ischemic neuronal death is associated with excitatory amino acid (EAA) release. Their action is mediated by N-methyl-D-aspartate (NMDA) receptors. Blockade of the receptors before the ischemic insult can decrease neuronal damage. Accordingly, we investigated the protective effect during spinal cord ischemia of two competitive antagonists, 4-(3-phosphonopropyl)-2-piperazine-carboxylic acid (CPP) and cis-4-(phosphonomethyl)-2-piperidine-carboxylic acid (CGS). Male Sprague-Dawley rats underwent intrathecal administration of 10 microL saline, CGS, and CPP 10 mM solutions, in a randomized blinded fashion, and were subjected to balloon occlusion of the thoracic aorta. Proximal aortic pressure was lowered to a mean of 40 mm Hg by partial exsanguination. In the acute protocol, 21 rats divided in 3 groups of 7 (saline, CPP, and CGS) were used to calculate the aortic occlusion time (AOT) resulting in paraplegia in 50% of animals (P50). In the chronic study, 24 rats divided in 4 groups of 6 (saline, CPP, CGS, sham) underwent 12-min occlusion. The chronic animals were scored daily for 28 days and submitted to histology of the cord. In the acute study, the P50 of CGS (10 min 48 s) and CPP (11 min 11 s) was longer than saline (10 min 27 s). In the chronic groups, analysis of variance of neurologic (p = .66) and histologic (p = .66) scores did not disclose differences between CGS, CPP, and saline. In conclusion, blockade of NMDA receptors with CPP or CGS may afford some protection for durations of occlusion around the P50, but it is not beneficial when ischemic injury is more protracted. PMID- 10801050 TI - Novel mutations in two Japanese cases of glycogen storage disease type IIIa and a review of the literature of the molecular basis of glycogen storage disease type III. AB - We report two novel mutations in two Japanese patients with glycogen storage disease type IIIa (GSD IIIa). In addition, we review the literature on mutations in GSD III to understand better the molecular basis of GSD III. In our first case, the homozygous A-to-C mutation at the acceptor site of intron 5 (IVS5-2A > C) was identified. This leads to the skipping of exon 6 and the predicted mutant protein was found to be 68 amino acids shorter than normal. This is the first report of skipping exon 6, which encodes one of the putative active sites, resulting in a profoundly deleterious effect on debrancher activity. In our second case, the homozygous deletion of an A at position 4234 (4234delA) was identified; this induces a frameshift resulting in the appearance of a stop codon at amino acid position 1276 (1276X). In patients with GSD IIIa, several mutations of the debrancher gene located in the C-terminal region containing putative glycogen binding domains have been identified as well as 4234delA in our second case. On the other hand, specific localization of the mutations within exon 3 was proposed in patients with GSD IIIb. PMID- 10801051 TI - Apolipoprotein E polymorphism and serum concentrations in patients with glycogen storage disease type Ia. AB - In patients with glycogen storage disease type Ia (glucose-6-phosphatase deficiency), serum triglyceride concentrations are markedly raised, whereas phospholipids and cholesterol levels are only moderately elevated. In addition, both VLDL and LDL lipoprotein fractions are raised. Despite these abnormalities, endothelial vascular dysfunction and atherosclerosis seem to be rare in such patients. In view of the crucial role of apolipoprotein E (apoE) in lipid metabolism, we studied both apoE polymorphism (40 patients) and serum concentration (20 patients) in patients with glycogen storage disease type Ia. The distribution of each allele at the apoE locus was similar to that reported in the general population, whereas serum apoE concentrations were raised in our patients. Raised apoE levels in the serum could play an important role in counterbalancing the at-risk-for-atherosclerosis lipid profile of patients with glycogen storage disease type Ia. Moreover, E3 and E4 polymorphisms, predominant in our patients, have a high triglyceride binding capacity and are thus able to increase triglyceride clearance. However, the origin of raised concentrations of apoE is not completely clear though, bearing in mind previous reports regarding serum protein concentrations in such patients, increased hepatic synthesis is likely. PMID- 10801052 TI - Enhanced lymphocyte proliferation in patients with adrenoleukodystrophy treated with erucic acid (22:1)-rich triglycerides. AB - Lymphocytopenia and depression of natural killer cells have been observed in patients with adrenoleukodystrophy (ALD) treated with glycerol trioleate and glycerol trierucate ('Lorenzo's oil'). To investigate possible alterations of cellular immunoreactivity, we measured lymphocyte proliferation in response to mitogens (PHA, Con A, PWM, OKT3) in 27 patients on treatment and in 14 patients without treatment. In patients on treatment, lymphocyte proliferation in response to the mitogens PHA and Con A was significantly higher than in patients without treatment. Lymphocyte proliferation in patients without treatment was comparable to that of normal control lymphocytes. Additionally, we found increased concentrations of erucic acid, C22:1, in lymphocytes from patients with treatment. The enhanced proliferation of lymphocytes in response to mitogens is an indication of increased reactivity of cellular immunity to unspecific immunological stimuli. Long-term side-effects on cellular immunoreactivity have to be considered in ALD patients treated with Lorenzo's oil. PMID- 10801053 TI - Novel mutations cause biotinidase deficiency in Turkish children. AB - Mutation analysis was performed on DNA from 31 Turkish children with profound biotinidase deficiency who were symptomatic or ascertained by newborn screening. The 98G:del7ins3 mutation is common in clinically ascertained children in both the United States and Turkish populations, but a unique common mutation, R79C, is found only in the Turkish children identified both clinically and by newborn screening. Another frequently occurring mutation, T532M, is only observed in the Turkish newborn screening group. There are four other less frequent novel mutations identified in the Turkish population. Interestingly, the Q456H and the A171T:D444H double mutation, which are the most common mutations found in the US newborn screening population and have not been observed in symptomatic children, do occur in clinically ascertained children in the Turkish population, although the double mutation may be associated with milder and/or later-onset symptoms. PMID- 10801054 TI - Three cases of intravenous sodium benzoate and sodium phenylacetate toxicity occurring in the treatment of acute hyperammonaemia. AB - Intravenous sodium benzoate and sodium phenylacetate have been used successfully in the treatment of acute hyperammonaemia in patients with urea cycle disorders. They provide alternative pathways for waste nitrogen disposal and help maintain nitrogen homeostasis. However, we report three patients with hyperammonaemia who received inappropriate doses of intravenous sodium benzoate and sodium phenylacetate that resulted in severe complications. Ambiguous medical prescriptions and inadequate cross-checking of drug dosage by physicians, nurses and pharmacists were the main causes of these incidents. All the patients presented with alteration in mental status, Kussmaul respiration and a partially compensated metabolic acidosis with an increased anion gap. Two patients developed cerebral oedema and hypotension and died. The third survived after haemodialysis. Plasma levels of benzoate and phenylacetate were excessively high. The possible mechanisms of toxicity, management and safety measures are discussed. PMID- 10801055 TI - Pulmonary hypertension associated with nonketotic hyperglycinaemia. AB - Nonketotic hyperglycinaemia (NKH) is an autosomal recessive disorder characterized by defective glycine degradation by the mitochondrial glycine cleavage system. The clinical features include lethargy, hypotonia, apnoea, seizures and severe psychomotor retardation, all attributed to the accumulation of glycine in the nervous system. Pulmonary hypertension (PHN) has not been reported in NKH. We describe four patients with NKH who had PHN in addition to the characteristic manifestations of NKH. This newly recognized association might provide additional insight into the underlying pathophysiology of PHN. PMID- 10801056 TI - Tripeptidyl-peptidase I deficiency in classical late-infantile neuronal ceroid lipofuscinosis brain tissue. Evidence for defective peptidase rather than proteinase activity. AB - Brain tissue from patients with classical late-infantile neuronal ceroid lipofuscinosis (LINCL, an infantile form of Batten disease) is deficient in the lysosomal enzyme tripeptidyl-peptidase I (EC 3.4.14.9). The activities of other lysosomal enzymes are either increased or decreased. Tripeptidyl-peptidase I is a pepstatin-insensitive exo-tripeptidase, with little or no endo-proteolytic activity, that is active on small peptides but not on large proteins. Using haemoglobin and casein as substrates for proteolytic activity, we were unable to demonstrate any significant defect in pepstatin-sensitive or pepstatin insensitive proteinase activity in brain tissue or cultured skin fibroblasts of LINCL patients. These observations suggest that the lysosomal storage of undegraded, small peptides in LINCL results from the absence of peptidase rather than proteinase activity. PMID- 10801057 TI - Multiple mtDNA deletions: clinical and molecular correlations. AB - We studied six Italian patients harbouring multiple mitochondrial DNA (mtDNA) deletions in order to correlate clinical and molecular features. Earlier age at onset (17 vs 36 years), fewer ragged-red fibres (none vs 35%), and lower proportions of deleted mtDNAs (9 vs 33%) were found in one patient with autosomal recessive inheritance as compared to five with dominant transmission. Our findings add to the features associated with multiple deletions of mtDNA. PMID- 10801058 TI - Genotypes and phenotypes of patients in the UK with carbohydrate-deficient glycoprotein syndrome type 1. AB - 18 UK patients (14 families) have been diagnosed with the carbohydrate-deficient glycoprotein syndrome (CDGS), type 1, on the basis of their clinical symptoms and/or abnormal electrophoretic patterns of serum transferrin. Eleven out of the 16 infants died before the age of 2 years. Patients from 12 families had a typical type 1 transferrin profile but one had a variant profile and another, who had many of the clinical features of CDGS type 1, had a normal profile. Eleven of the patients (10 families) with the typical type 1 profile had a deficiency of phosphomannomutase (PMM), (CDGS type 1a) but there was no correlation between residual enzyme activity and severity of disease. All these patients were compound heterozygotes for mutations in the phosphomannomutase (PMM2) gene, with 7 out of the 10 families having the common R141H mutation. Eight different mutations were found, including three novel ones. There was no correlation between genotype and phenotype, although siblings had similar phenotypes. Three patients, including the one with the normal transferrin profile, did not have a deficiency of phosphomannomutase or phosphomannose isomerase (CDGS 1b). PMID- 10801059 TI - The increased sensitivity of neurons with elevated glucocerebroside to neurotoxic agents can be reversed by imiglucerase. AB - We have recently demonstrated that incubation of cultured rat hippocampal neurons with conduritol beta-epoxide (CBE), an inhibitor of glucocerebrosidase, the enzyme defective in Gaucher disease, results in changes in intracellular morphology and in functional calcium stores. Changes in levels of functional calcium stores are directly related to neuronal cell death. We now show that neurons incubated with either CBE or a non-hydrolysable analogue of GlcCer (glucosylthioceramide), are more sensitive to the toxic effects of high concentrations of glutamate and of a variety of metabolic inhibitors. A linear relationship exists between level of accumulation of GlcCer and the extent of neuronal cell death. The deleterious effects of elevated GlcCer levels can be completely reversed by addition of human glucocerebrosidase (imiglucerase) to the culture medium. Imiglucerase is internalized to lysosomes, where it presumably degrades excess GlcCer. This suggests that the limited success of enzyme replacement therapy in neuronopathic forms of Gaucher disease is not due to lack of efficacy of glucocerebroside in degrading GlcCer in neurons of the central nervous system, and adds impetus to attempts to develop ways to efficiently deliver glucocerebrosidase to the brains of neurologically compromised Gaucher disease patients. PMID- 10801060 TI - Novel mutations in children with profound biotinidase deficiency from Saudi Arabia. PMID- 10801061 TI - Long-chain hydroxydicarboxylic aciduria, carnitine depletion and acetaminophen exposure. PMID- 10801062 TI - Acute pancreatitis in homocystinuria. PMID- 10801063 TI - Development of a parent questionnaire for use in assessing adolescent drug abuse. AB - The development and initial psychometric properties of a new parent questionnaire for assessing adolescent drug abuse are described. The Personal Experience Inventory-Parent Version (PEI-PV) is intended to provide a standardized parent report as a companion measure to the adolescent self-report instrument, the Personal Experience Inventory. The PEI-PV addresses problems associated with the child's drug abuse and psychosocial factors and parenting practices that may underlie their drug involvement. Reliability and validity data are reported for two groups of mothers, 205 of whom had a child referred for an evaluation for drug treatment and 185 from a community sample. The structure and scope of the PEI-PV was supported by scale intercorrelation data; most scales had a proportion of unique, reliable variance greater than 20%. Estimates of the scale's internal consistency were found to be comparable to those of established parent questionnaires, and the observed differences between the clinic-referred and community samples were consistent with expectations. Significant convergence of mothers' reports to those of their child was obtained for nearly three-quarters of the PEI-PV scales, including moderate agreement between mother and child on the drug involvement severity scales. However, mothers tended to underreport their child's level of drug involvement and resulting problems compared to the child's self-report. The study results suggest that the PQ is associated with favorable evidence that the scales measure what they are intended to measure. PMID- 10801064 TI - Social influences as explanations for substance use differences among Asian American and European-American adolescents. AB - The present study examines the social influences in the Asian-American and European-American adolescent environment to provide a greater understanding of effect of ethnic differences on the prevalence of substance abuse. Participants were 957 Asian-American and 3705 European-American seventh grade adolescents. It was found that Asian-American students were less likely to use alcohol and cigarettes, and had a more abstinence-promoting environment than European American adolescents. This difference in Asian-American adolescents appeared to be the result of less adult and peer influence to use alcohol or cigarettes, less offers of alcohol, and an increased likelihood of having an intact family when compared to European-American adolescents. Additionally, Asian-American adolescents reported having fewer friends and spending less time with their friends than European-American adolescents. It was concluded that social influences play an important role in explaining why Asian-American adolescents have lower rates of substance use. PMID- 10801065 TI - Understanding gender differences in adolescent drug abuse: issues of comorbidity and family functioning. AB - Female adolescent drug use has increased dramatically in the last 30 years, and there is a growing consensus that the syndrome of female adolescent substance abuse is different from the well-recognized male pattern. Gender differences in patterns of comorbidity and family functioning were investigated in a sample of 95 youths (42 girls and 53 boys) referred for substance abuse treatment. The findings indicate that male and female adolescent substance users differ in several clinically meaningful ways. The results from a discriminant function analysis indicate that substance-using adolescents referred to treatment are distinguished especially by the greater degree to which girls have internalizing symptoms and family dysfunction. The clinical implications of these gender differences are articulated. PMID- 10801066 TI - The role of acculturation in the substance abuse behavior of African-American and Latino adolescents: advances, issues, and recommendations. AB - This article provides a review of the literature on the acculturation of Latino and African-American adolescents to traditional European-American cultural values, and the effect on their substance abusing behaviors. The review includes a critical analysis of studies that examine the effects of acculturation on the mental health and well-being of Latino adolescents. Recent findings documenting the association between acculturation and substance abuse among Latino adolescents are discussed. The article also examines the dearth of research on the role of acculturation in the substance-abusing behaviors of African-American adolescents. The authors assert that understanding the effects of acculturation on these behaviors could begin to explain why African-American adolescent substance abuse rates have been historically lower than those of European American and Latino adolescents. Also included is an examination of research on the role of acculturation in treatment outcomes of Latino and African-American adolescents. PMID- 10801067 TI - Trauma, drugs and violence among juvenile offenders. AB - Trauma typically occurs when one experiences a situation where life has been threatened or lost. If the trauma is not resolved, negative residual effects may result in alcohol and drug use, involvement in violent activities as well as the development of mental health problems such as posttraumatic stress disorder (PTSD). Findings from a National Institute on Drug Abuse-funded study examining the link between trauma, drug use and violence among youth are presented. Results from interviews with 414 juveniles remanded to the Office of Children and Family Services (formerly New York State Division For Youth) for assault, sexual assault, robbery or homicide, document the trauma experienced by these youth, as well as how it correlated with their drug usage and participation in violent, illegal activities. Discussion of these findings, their implications for understanding and intervening, and recommendations for future research are highlighted. PMID- 10801068 TI - Testing a model of the influence of family problem factors on high-risk youths' troubled behavior: a three-wave longitudinal study. AB - Using data collected in a study of arrested youths processed at a juvenile assessment center who entered a home-based family intervention project, the authors test a three-stage longitudinal model. The model reflects a developmental damage view of the relationships of the youths' family problems (including their physical abuse and sexual victimization experiences) to their drug use and delinquent behavior over time. The hypothesized model is supported by the data. Research, theoretical, and policy implications of the results are drawn. PMID- 10801069 TI - Disruptive behavior disorders and substance use disorders in adolescents. AB - Disruptive behaviors disorders in the form of conduct disorder, oppositional defiant disorder and/or attention-deficit hyperactivity disorder are found in a majority of adolescents with substance use disorders These disorders influence the risk for and the course of substance use disorders in adolescents and potentially provide important targets for intervention. Interventions such as family therapy and multisystemic therapy can focus on important environmental factors that help to produce and sustain substance use, related problems and disruptive/deviant social behavior. Researchers and clinicians are increasingly utilizing multimodal approaches that use several psychosocial approaches in addition to medication, if indicated. This article reviews our current understanding of the relationship between disruptive behavior disorders and substance use disorders in adolescents and the importance of this understanding in the prevention, assessment and treatment of adolescents with substance use disorders. PMID- 10801070 TI - Adolescents in therapeutic communities: one-year posttreatment outcomes. AB - Given the dearth of information concerning outcomes for adolescents who have been in treatment for substance abuse and related problems, the purpose of the present article is to examine the posttreatment status of adolescents who have been in residential therapeutic communities (TCs). One-year posttreatment outcome data are described for 485 adolescents, and separately for treatment "completers" and "noncompleters." The majority of the sample had been mandated to treatment by the criminal justice system. Most reported marijuana as their main drug of abuse. There were significant reductions in drug use and criminal activity, and the most consistent predictors of positive outcomes were completion of treatment and not associating with deviant peers posttreatment. The findings were obtained on a treatment sample for whom there is relatively little research to date; they provide important evidence for the effectiveness of the therapeutic community for this population. PMID- 10801071 TI - Adolescent amphetamine users in treatment: client profiles and treatment outcomes. AB - This study compares client profiles of adolescent amphetamine users to those of nonusers and examines the multivariate prediction of posttreatment drug use, criminal, and HIV risk behavior outcomes in the year following their separation from treatment. Data were collected as part of a larger longitudinal study on a sample of 938 adolescents who were admitted to residential therapeutic community drug treatment programs across the eastern United States and Canada from April of 1992 through April of 1994. A subsample of 485 adolescents were reinterviewed one year after their separation from treatment. Findings indicated that amphetamine users tended to be white, older, and have parents with higher education and occupational levels than nonusers. However, they also had more psychopathology, more extensive drug use and criminal histories, and engaged in more HIV-risk behaviors than nonusers. Additionally, amphetamine users tend to come from homes where one or both parents used illicit drugs, drank regularly, or had a mental illness, and they often reported histories of childhood maltreatment. Analyses of the one-year follow-up data indicated that being an amphetamine user was not related to treatment outcome after the client's demographic characteristics, overall drug use severity, and treatment completion were taken into account. Therefore, therapeutic community treatment appears to be an effective means of treating adolescent amphetamine users. PMID- 10801072 TI - The role of alcohol in adolescent relapse and outcome. AB - Although adolescents entering treatment for substance abuse typically use several substances, alcohol is seldom the drug of choice. Given alcohol's role as a gateway substance in the initiation of substance misuse, the authors hypothesize that alcohol is substantially involved in adolescent relapse following substance abuse treatment. One hundred fifty-seven youths (39.5% female, mean age 15.91 years) were recruited from adolescent inpatient alcohol and drug treatment centers and assessed at six and 12 months after discharge. Of the youths studied, 79% had used one or more substances by 12-month follow-up. Although only 1% reported alcohol as their substance of choice while hospitalized, alcohol was involved in 46% of initial posttreatment use episodes. Almost a quarter (23%) of initial posttreatment substance use events involved multiple substances. Initial posttreatment use episodes involving multiple substances and preferred substances were associated with poorer outcomes during the year following treatment both in terms of rate of return to and severity of substance involvement. Youths who initially used only alcohol were also likely to resume harmful levels of substance involvement by one year after treatment. Results suggest two routes, gateway and preferred substance, for adolescents in the resumption of harmful substance use following treatment. This gateway role merits consideration by adolescent substance abuse treatment providers and families of treated youths. PMID- 10801073 TI - A retrospective look at long-term adolescent recovery: clinicians talk to researchers. AB - The purpose of the present article is to identify factors associated with long term recovery for adolescents. A group of 14 young adults were given semistructured in-depth (one to two hour) interviews. The subjects had been in recovery for two to 16 years. Length of stay in treatment, involvement with self help groups, psychotropic medication, family support, peer support, spirituality, and other factors were identified as helpful for long-term recovery. A major limitation of this inquiry is that it does not meet minimum standards for research. It is hoped, however, that this inquiry can inform directions for further research. PMID- 10801074 TI - Role of the Bsk/Iyk non-receptor tyrosine kinase for the control of growth and hormone production in RINm5F cells. AB - Bsk/Iyk, a murine non-receptor-tyrosine kinase which is expressed in fetal and adult islet of Langerhans was previously found to decrease NIH3T3 cell proliferation when expressed as a Y497/504F-mutant. We presently wanted to determine the effects of Bsk/Iyk on the proliferation of insulin producing cells. Cells expressing Bsk/IykY497/504F and Bsk/IykY504F display a decreased proliferation rate and express higher levels of the cell cycle inhibitor p27/Kip1 compared to control cells. These mutants also conferred diminished cell viability in response to INF-gamma and IL-1beta and contain higher levels of glucagon mRNA. Wild-type Bsk/Iyk is mainly localized at the plasma membrane whereas mutant Bsk/Iyk can enter the nucleus. In vitro kinase reactions using an exogenous substrate indicate a complicated mode of regulation of kinase activity by Y497 and Y504 with the latter being homologous to Y527 in pp60c-Src. These findings suggest that Bsk/Iyk might play a role in inhibiting cell proliferation, transducing cytokine-induced cytotoxicity and regulating hormone production of endocrine pancreatic cells. PMID- 10801075 TI - Gene response of human skin fibroblasts to urokinase- and tissue-type plasminogen activators. AB - In a previous work we have reported evidences on the mitogenic activity of urokinase-type and tissue-type plasminogen activator (u-PA, t-PA) on serum deprived human dermal fibroblasts. In this work we have studied the transcription dependent changes of some cell-cycle related genes associated with the biological activity of PAs, as well as the possible involvement of protein tyr kinases (PTK) and/or protein kinase C (PKC) in the mitogenic signal transduction. The data obtained demonstrate that the growth factor activity of PAs is associated with: - a rapid transient activation of early response genes, c-fos, c-jun and c-myc; - the subsequent coordinated down-regulation of p53 and p21CIP1; - the constant expression of the MEK1 mRNA in every phase of the cell cycle. Quiescent (G0) cells did not express c-fos, c-jun, c-myc and cyclin A, but upon stimulation with mitogens (fetal calf serum (FCS), u-PA, t-PA) the cyclin A mRNA expression was observed in concomitance with the activation of DNA synthesis. Therefore u-PA, t PA and FCS similarly modulate the expression of c-fos, c-jun, c-myc, p53, p21CIP1 and cyclin A with only slight differences likely related to the time required for activation of DNA synthesis. The PAs mitogenic stimulation of serum-starved cells was associated with the internalization of their molecules, as revealed by immunostaining. The biological activity of u-PA, t-PA, as well as that of limiting concentration of FCS (1%), was mediated by PTK and PKC. Conversely, PTK, but not PKC, was involved in the activation of the proliferative response of basic fibroblast growth factor in the same experimental conditions. In conclusion, u-PA and t-PA can utilize two different pathways, one depending on PTK and the other on PKC in a way similar to the mitogenic activity induced by low concentration of FCS (1%). PMID- 10801076 TI - Induction of endochondral bone formation by recombinant human transforming growth factor-beta2 in the baboon (Papio ursinus). AB - Members of the transforming growth factor-beta (TGF-beta) superfamily, the bone morphogenetic and osteogenic proteins (BMPs/OPs) but not the TGF-beta proteins themselves, induce endochondral bone formation in vivo, when implanted in extraskeletal heterotopic sites of rodents. Here we show that recombinant human TGF-beta2 (hTGF-beta2) induces endochondral bone formation 30 days after implantation in heterotopic intramuscular sites of the baboon (Papio ursinus) at doses of 1, 5 and 25 microg per 100 mg of guanidinium-inactivated collagenous bone matrix as carrier. On day 90 there was generation of large radiopaque and corticalized intramuscular ossicles. Five and 25 microg hTGF-beta2 induced large ossicles in the rectus abdominis of the primate as evaluated by key parameters of bone formation, including generated tissue area, mineralized bone and osteoid volumes, and tissue alkaline phosphatase activity. On day 30 and 90 after healing, hTGF-beta2 also induced bone formation when implanted in the rectus abdominis in conjunction with a sintered porous hydroxyapatite as carrier. mRNA expression in tissues from heterotopic specimens showed OP-1 (BMP-7) and BMP-3 transcripts in low abundance and with a linear dose-dependent increase both in collagenous matrix and hydroxyapatite samples. Type IV collagen mRNA expression, a marker of angiogenesis, was stronger in collagenous than hydroxyapatite samples. Growth and differentiation factor-10 (GDF-10) mRNA transcripts were expressed in ossicles with a distinctly chondrogenic phase, but its expression was greater in ossicles generated in porous hydroxyapatites, in which bone formation is not via a chondrogenic phase, but is rather intramembranous, without expression of type II collagen mRNA. In the same animals, however, 10 and 100 microg of the recombinant morphogen delivered by identical carriers (collagenous matrix and sintered hydroxyapatite) failed to heal calvarial defects. Thus in the primate, TGF-betas themselves are inducers of endochondral bone formation, although the present data strongly indicate that the bone inductive activity of hTGF-beta2 is site and tissue specific, since a single application of hTGF-beta2, or hTGF-beta1 in previously published experiments, did not induce bone in calvarial defects, but did induce endochondral bone differentiation in heterotopic sites. PMID- 10801077 TI - A phase I and pharmacokinetic study of subcutaneously-administered recombinant human interleukin-4 (rhuIL-4) in patients with advanced cancer. AB - PURPOSE: To investigate the pharmacokinetics and tolerability of recombinant human interleukin-4 (rhuIL-4), administered by daily subcutaneous injection, in patients with advanced cancer. PATIENTS AND METHODS: Fourteen patients with advanced cancer treated with rhuIL-4 at escalating dose levels of 0.25, 1.0 and 5.0 microg/kg/day, on days 1, 8-17, and 28-57. The primary endpoints of the study were toxicity of rhuIL-4 and the determination of the pharmacokinetics of rhuIL-4 when given by subcutaneous injection. Secondary endpoints included effects on blood counts, hematopoietic cell precursors, and various immunologic parameters. RESULTS: rhuIL-4 was well tolerated at all three dose levels. Detectable serum levels of IL-4 were found in patients at the 1.0 and 5.0 microg/kg/day dose levels. Peak serum IL-4 levels were achieved about 2 h after injection and IL-4 was still detectable 8 h after injection. No grade 4 toxicities were observed and grade 3 toxicities were confined to fever, headache and raised hepatic alkaline phosphatase. No consistent hematological or immunologic effects were observed. Although therapeutic efficacy was not an endpoint, one complete response (Hodgkin's disease) was observed. One patient with chronic lymphocytic leukemia progressed on therapy. CONCLUSION: rhuIL-4 up to 5.0 microg/kg/day is well tolerated when given by subcutaneous injection. Biologically relevant serum IL-4 levels can be achieved and sustained for at least 8 h after a single injection. PMID- 10801078 TI - The role of vascular endothelial growth factor (VEGF) in abnormal vascular changes in the adult rat eye. AB - The aim of this project was to determine if the subretinal delivery of a recombinant adenovirus encoding vascular endothelial growth factor (VEGF) was sufficient to induce changes resembling choroidal neovascularisation (CNV) in a rat model. A recombinant adenovirus was produced encoding vegf164 cDNA (Ad.RSV.VEGF). Transduction of cultured RPE cells confirmed VEGF expression and ensured the absence of Ad.RSV.VEGF-related toxicity. Following subretinal injection into rat eyes, fluorescein angiography indicated that the in vivo delivery of Ad.RSV.VEGF was associated with vascular leakage. Histological analysis demonstrated that changes resembling the early signs of CNV development were also present in the Ad.RSV.VEGF injected eyes. These results suggest that while a transient VEGF expression in the RPE layer is able to induce CNV-related changes, it may be insufficient for the development of a full neovascular membrane. This study demonstrates that virus-mediated gene delivery, in addition to its clinical applications, is a potentially efficient research tool for investigating gene expression-related physiological changes in vitro and in vivo. PMID- 10801079 TI - Measuring consistency of web page design and its effects on performance and satisfaction. AB - This study examines the methods for measuring the consistency levels of web pages and the effect of consistency on the performance and satisfaction of the world wide web (WWW) user. For clarification, a home page is referred to as a single page that is the default page of a web site on the WWW. A web page refers to a single screen that indicates a specific address on the WWW. This study has tested a series of web pages that were mostly hyperlinked. Therefore, the term 'web page' has been adopted for the nomenclature while referring to the objects of which the features were tested. It was hypothesized that participants would perform better and be more satisfied using web pages that have consistent rather than inconsistent interface design; that the overall consistency level of an interface design would significantly correlate with the three elements of consistency, physical, communicational and conceptual consistency; and that physical and communicational consistencies would interact with each other. The hypotheses were tested in a four-group, between-subject design, with 10 participants in each group. The results partially support the hypothesis regarding error rate, but not regarding satisfaction and performance time. The results also support the hypothesis that each of the three elements of consistency significantly contribute to the overall consistency of a web page, and that physical and communicational consistencies interact with each other, while conceptual consistency does not interact with them. PMID- 10801080 TI - Pointing movement visually controlled through a video display: adaptation to scale change. AB - This study concerns the adaptation of motor system when the production of movement was visually controlled through a video display. The subjects had to perform a pointing movement in two visual-feedback conditions: they could see the displacement of their hand or only the final location of the hand on the videoscreen. By changing the zoom of the camera, the amplitude of the movement perceived on the screen was increased, decreased or held equal to the actual movement. Results showed that the movement adapted quickly to the apparent distance changes. In the full visual-feedback condition, the adaptation was smaller in magnitude than in the partial visual-feedback condition. Even though the actual movement was always the same, the subjects thought they carried out different movements. Therefore, the subjects did not use kinematic information provided by the kinaesthetic system but essentially visual information furnished by the video-screen. Taken together, these results show that adaptation to scale changes does not allow the achievement of the perception of a single working space but seems rather specific to each scale. PMID- 10801081 TI - Positive expiratory pressure as a method for preventing the impairment of attentional processes by hypoxia. AB - This study investigated the effects of hypoxia on parallel/preattentional and serial/attentional processes in early vision, and the use of a positive-end expiratory-pressure (PEEP) to prevent the impairment in performance. Twenty-one subjects were submitted to an 8-h hypoxia exposure in a hypobaric chamber (4500 m, 589 hPa, 22 degrees C), both with and without a 5-cm H2O PEEP. Subjects carried out a visual search task consisting of detecting a target among distractors in normoxia, in acute and in prolonged hypoxia. Conjointly their sensitivity to acute mountain sickness (AMS) was scored through the Lake Louise AMS scoring system. Results showed that prolonged hypoxia slowed serial/attentional processing whereas parallel/preattentional processes were not impaired either by acute or by prolonged hypoxia. PEEP prevented serial/attentional processes from slowing and those effects were more clearly observed in the AMS sensitive subjects with respect to the AMS insensitive subjects. These results suggest that the slowing induced by prolonged hypoxia is specific to an early visual process that pilots the scanning of an attentional spotlight throughout the visual field. PMID- 10801083 TI - Training in virtual environments: transfer to real world tasks and equivalence to real task training. AB - Virtual environments (VEs) are extensively used in training but there have been few rigorous scientific investigations of whether and how skills learned in a VE are transferred to the real world. This research aimed to measure and evaluate what is transferring from training a simple sensorimotor task in a VE to real world performance. In experiment 1, real world performances after virtual training, real training and no training were compared. Virtual and real training resulted in equivalent levels of post-training performance, both of which significantly exceeded task performance without training. Experiments 2 and 3 investigated whether virtual and real trained real world performances differed in their susceptibility to cognitive and motor interfering tasks (experiment 2) and in terms of spare attentional capacity to respond to stimuli and instructions which were not directly related to the task (experiment 3). The only significant difference found was that real task performance after training in a VE was less affected by concurrently performed interference tasks than was real task performance after training on the real task. This finding is discussed in terms of the cognitive load characteristics of virtual training. Virtual training therefore resulted in equivalent or even better real world performance than real training in this simple sensorimotor task, but this finding may not apply to other training tasks. Future research should be directed towards establishing a comprehensive knowledge of what is being transferred to real world performance in other tasks currently being trained in VEs and investigating the equivalence of virtual and real trained performances in these situations. PMID- 10801082 TI - Lumbar erector spinae oxygenation during prolonged contractions: implications for prolonged work. AB - Owing to the recent interest in torso stabilization exercises together with many questions regarding the duration of prolonged isometric holds in occupational settings, the authors attempted to assess the level of back muscle oxygenation during prolonged isometric contractions. Specifically, this study recorded relative oxygen saturation of haemoglobin/myoglobin using Near Infrared Spectroscopy (NIRS) in the L3 erector mass during prolonged isometric contractions at intensities from 2 to 30% of maximum voluntary contraction (MVC). It was hypothesized that available oxygen to these muscles is severely compromised even at moderate levels of activation observed in occupational work. Eight volunteers without a history of lower back pain or injury participated in this study. The exercise task involved isometric contraction of the lower erector spinae at five different levels of each subject's maximal voluntary contraction: 2, 5, 10, 20 and 30% MVC, presented in random order. Subjects were placed in a sitting position, with a curved plastic plate secured horizontally to the pelvis to minimize movement at the hip joint. During extensor exertions, they were restrained with a harness that was attached at chest level to a load cell. Each isometric contraction was performed for 30 s followed by 1 min of rest. All levels of contraction demonstrated reduction in oxygen. Given the concern for motion artefact on the NIRS signal, sham trials were conducted where the subjects went through the procedure of attaching the pulling cable but no active pull was performed. These trials showed no change in the NIRS signal. At this time NIRS appears to be the only non-invasive instrumentation available to indicate total available muscle oxygen during low level, prolonged work. Although the specific tissue volume sampled by NIRS cannot be positively identified, it appears that tissue oxygenation in the lumbar extensor musculature is reduced as a function of contraction intensity, even at levels as low as 2% of MVC. These data have implications for prolonged work where postures requiring isometric contractions are sometimes held for hours, and where musculoskeletal illness has been linked to prolonged contraction levels above 2%MVC--these data suggest a possible biological pathway. PMID- 10801084 TI - Effect of trunk rotation and arm position on gross upper extremity adduction strength and muscular activity. AB - The aim of the experiment was to determine the impact of axial trunk rotation and arm position on upper extremity adduction force and muscle activity. Ten healthy male subjects performed graded maximum voluntary contractions under isometric conditions in seven upper extremity positions and three trunk postures (neutral and 90 degrees left/right rotated) in a simulated manual materials handling task. A custom built lightweight force-measuring device was held between the palmar surfaces of the hands and subjects compressed the lateral surfaces of the device. Muscle activity was recorded bilaterally over the muscle bellies of the anterior deltoid, the long head of the biceps brachii and over the flexor carpi radialis. The activity of the right pectoralis major was also recorded unilaterally. Descriptive, multivariate analysis of variance (MANOVA) and post-hoc Scheffe comparisons were performed on the mean and peak force as well as the EMG [electromyographic] data. Further analysis was performed on the force-EMG relationship at 20% intervals of maximum voluntary contraction (force). Both upper extremity adduction force and EMG were significantly affected by position (p<0.01) but not by trunk rotation. The muscle activity increased and force decreased with flexion of the upper extremity. Pearson correlation coefficients between force and EMG were low. The biceps and flexors were the most active muscles depending upon upper extremity position, and the right pectoralis major muscle activity expressed the highest correlation with force. The present findings confirm earlier hypotheses that upper extremity adduction strength is not significantly affected by trunk rotation. PMID- 10801085 TI - Examination of the use of fuzzy sets to describe relative speed perception. AB - In recent years a range of new methods have been proposed with which to describe and evaluate driver behaviour. One such method is that of fuzzy logic, where variables used in the driver decision-making process may be described linguistically, allowing a quantifiable degree of uncertainty to be introduced. This paper explores the use of such a formalism to describe the driver perception of 'closing speed' between two vehicles engaged in 'car-following' on a motorway, and by using data from an instrumented vehicle experiment, it tests a number of models using relative speed, visual angle and the time to collision. Several of these models fit the data quite well, and there is both a small positive perception bias present and a number of reversals in sign judgement. Additionally, a brief examination is made of potential variations on the methodology that may both make data collection easier and/or allow a 'more fuzzy' representation to be made. PMID- 10801086 TI - Designing safe job rotation schedules using optimization and heuristic search. AB - Job rotation is one method that is sometimes used to reduce exposure to strenuous materials handling; however, developing effective rotation schedules can be complex in even moderate sized facilities. The purpose of this research is to develop methods of incorporating safety criteria into scheduling algorithms to produce job rotation schedules that reduce the potential for injury. Integer programming and a genetic algorithm were used to construct job rotation schedules. Schedules were comprised of lifting tasks whose potential for causing injury was assessed with the Job Severity Index. Each method was used to design four job rotation schedules that met specified safety criteria in a working environment where the object weight, horizontal distance and repetition rate varied over time. Each rotation was assigned to a specific gender/lifting capacity group. Five versions of the integer programming search method were applied to this problem. Each version generated one job rotation schedule. The genetic algorithm model was able to create a population of 437 feasible solutions to the rotation problem. Utilizing cluster analysis, a rule set was derived from the genetic algorithm generated solutions. These rules provided guidelines for designing safe job rotation schedules without the use of a computer. The advantages and limitations of these approaches in developing administrative controls for the prevention of back injury are discussed. PMID- 10801088 TI - Big can still be great PMID- 10801089 TI - Budget crisis forces hard choices on US high-energy physics PMID- 10801087 TI - Dear Mr Mbeki... PMID- 10801090 TI - 'Saturation screen' lets zebrafish show their stripes. PMID- 10801091 TI - Letter fuels South Africa's AIDS furore. PMID- 10801093 TI - NASA's new environmental satellite shows its sensitive side PMID- 10801092 TI - Genetic diversity project fights for its life...as companies are urged to share benefits. PMID- 10801094 TI - Japan gears up for growth in genomics. PMID- 10801095 TI - New Zealand GM inquiry will cast a wide net. PMID- 10801096 TI - Taiwan's kingmaker chemist. PMID- 10801097 TI - Meet the spin doctors... PMID- 10801098 TI - How diagnosis with microarrays can help cancer patients. PMID- 10801099 TI - Short-sighted move to close the 12-m telescope PMID- 10801100 TI - Genetic modification and the meat market. PMID- 10801101 TI - Distinguished scientists back Germany's DFG... PMID- 10801102 TI - But young researchers feel disillusioned. PMID- 10801103 TI - 'Marketing' species conservation. PMID- 10801104 TI - Putting technology in its place PMID- 10801105 TI - The enduring test PMID- 10801106 TI - Ringing in the new cosmology PMID- 10801107 TI - Molecular biology. Introns gain ground. PMID- 10801108 TI - Heat flow through nanobridges PMID- 10801109 TI - Huntington's disease. In reverse gear. PMID- 10801111 TI - Superconducting nanowires PMID- 10801110 TI - Molecular switches in lipid rafts. PMID- 10801113 TI - Focusing hard X-rays with old LPs PMID- 10801112 TI - Decoy receptors thwart B cells. PMID- 10801114 TI - Symbiotic solution to arsenic contamination. PMID- 10801115 TI - Tunes and templates. PMID- 10801116 TI - Palaeontology. A refugium for relics? PMID- 10801117 TI - A flat Universe from high-resolution maps of the cosmic microwave background radiation AB - The blackbody radiation left over from the Big Bang has been transformed by the expansion of the Universe into the nearly isotropic 2.73 K cosmic microwave background. Tiny inhomogeneities in the early Universe left their imprint on the microwave background in the form of small anisotropies in its temperature. These anisotropies contain information about basic cosmological parameters, particularly the total energy density and curvature of the Universe. Here we report the first images of resolved structure in the microwave background anisotropies over a significant part of the sky. Maps at four frequencies clearly distinguish the microwave background from foreground emission. We compute the angular power spectrum of the microwave background, and find a peak at Legendre multipole Ipeak = (197 +/- 6), with an amplitude delta T200 = (69 +/- 8) microK. This is consistent with that expected for cold dark matter models in a flat (euclidean) Universe, as favoured by standard inflationary models. PMID- 10801118 TI - Structure of the reovirus core at 3.6 A resolution. AB - The reovirus core is an assembly with a relative molecular mass of 52 million that synthesizes, modifies and exports viral messenger RNA. Analysis of its structure by X-ray crystallography shows that there are alternative, specific and completely non-equivalent contacts made by several surfaces of two of its proteins; that the RNA capping and export apparatus is a hollow cylinder, which probably sequesters its substrate to ensure completion of the capping reactions; that the genomic double-stranded RNA is coiled into concentric layers within the particle; and that there is a protein shell that appears to be common to all groups of double-stranded RNA viruses. PMID- 10801119 TI - Identification of molecular-cloud material in interplanetary dust particles. AB - Interplanetary dust particles (IDPs) collected in the Earth's stratosphere and meteorites are fragments of comets and asteroids. These are 'primitive' meteorites in part because they have preserved materials which predate the formation of the Solar System. The most primitive (least altered) meteorites contain a few parts per million of micrometre-sized dust which formed in the atmospheres of giant stars. Some meteorites have elevated D/H and 15N/14N ratios that are attributed to surviving interstellar organic molecules which have probably been strongly diluted and altered by parent-body processes. Most IDPs are chemically, mineralogically, and texturally primitive in comparison to meteorites. Here I show that H and N isotopic anomalies among fragile 'cluster' IDPs are far larger, more common, and less equilibrated than those previously observed in other IDPs or meteorites. In some cases, the D/H ratios that we measure reach the values of interstellar molecules, suggesting that molecular cloud material has survived intact. These observations indicate that cluster IDPs are the most primitive class of Solar System materials currently available for laboratory analysis. PMID- 10801120 TI - Quantum suppression of superconductivity in ultrathin nanowires AB - It is of fundamental importance to establish whether there is a limit to how thin a superconducting wire can be, while retaining its superconducting character--and if there is a limit, to determine what sets it. This issue may also be of practical importance in defining the limit to miniaturization of superconducting electronic circuits. At high temperatures, the resistance of linear superconductors is caused by excitations called thermally activated phase slips. Quantum tunnelling of phase slips is another possible source of resistance that is still being debated. It has been theoretically predicted that such quantum phase slips can destroy superconductivity in very narrow wires. Here we report resistance measurements on ultrathin (< or = 10 nm) nanowires produced by coating carbon nanotubes with a superconducting Mo-Ge alloy. We find that nanowires can be superconducting or insulating depending on the ratio of their normal-state resistance (R(N)) to the quantum resistance for Cooper pairs (Rq). If R(N) < Rq, quantum tunnelling of phase slips is prohibited by strong damping, and so the wires stay superconducting. In contrast, we observe an insulating state for R(N) > Rq, which we explain in terms of proliferation of quantum phase slips and a corresponding localization of Cooper pairs. PMID- 10801121 TI - Measurement of the quantum of thermal conductance AB - The physics of mesoscopic electronic systems has been explored for more than 15 years. Mesoscopic phenomena in transport processes occur when the wavelength or the coherence length of the carriers becomes comparable to, or larger than, the sample dimensions. One striking result in this domain is the quantization of electrical conduction, observed in a quasi-one-dimensional constriction formed between reservoirs of two-dimensional electron gas. The conductance of this system is determined by the number of participating quantum states or 'channels' within the constriction; in the ideal case, each spin-degenerate channel contributes a quantized unit of 2e(2)/h to the electrical conductance. It has been speculated that similar behaviour should be observable for thermal transport in mesoscopic phonon systems. But experiments attempted in this regime have so far yielded inconclusive results. Here we report the observation of a quantized limiting value for the thermal conductance, Gth, in suspended insulating nanostructures at very low temperatures. The behaviour we observe is consistent with predictions for phonon transport in a ballistic, one-dimensional channel: at low temperatures, Gth approaches a maximum value of g0 = pi2kB2T/3h, the universal quantum of thermal conductance. PMID- 10801122 TI - Coulomb-blockade transport in single-crystal organic thin-film transistors AB - Coulomb-blockade transport--whereby the Coulomb interaction between electrons can prohibit their transport around a circuit--occurs in systems in which both the tunnel resistance, Rb between neighbouring sites is large (>>h/e2) and the charging energy, E(C) (E(C) = e2/2C, where C is the capacitance of the site), of an excess electron on a site is large compared to kT. (Here e is the charge of an electron, k is Boltzmann's constant, and h is Planck's constant.) The nature of the individual sites--metallic, superconducting, semiconducting or quantum dot- is to first order irrelevant for this phenomenon to be observed. Coulomb blockade has also been observed in two-dimensional arrays of normal-metal tunnel junctions, but the relatively large capacitances of these micrometre-sized metal islands results in a small charging energy, and so the effect can be seen only at extremely low temperatures. Here we demonstrate that organic thin-film transistors based on highly ordered molecular materials can, to first order, also be considered as an array of sites separated by tunnel resistances. And as a result of the sub-nanometre sizes of the sites (the individual molecules), and hence their small capacitances, the charging energy dominates at room temperature. Conductivity measurements as a function of both gate bias and temperature reveal the presence of thermally activated transport, consistent with the conventional model of Coulomb blockade. PMID- 10801123 TI - Inorganic yellow-red pigments without toxic metals AB - Inorganic pigments have been utilized by mankind since ancient times, and are still widely used to colour materials exposed to elevated temperatures during processing or application. Indeed, in the case of glasses, glazes and ceramics, there is no alternative to inorganic pigments for colouring. However, most inorganic pigments contain heavy metals or transition metals that can adversely effect the environment and human health if critical levels are exceeded. Cadmium based pigments in particular are a cause of concern: although the pigments are not toxic due to their very low solubility in water and dilute mineral acids, cadmium itself is toxic and can enter the environment in a bioavailable form through waste-disposal sites and incineration plants. This has led to regulations, based on the precautionary principle, that strongly restrict the use of cadmium pigments. And even though recent assessments have concluded that the risk to humans or the environment might be not as significant as originally feared, a strong demand for inherently safer substitutes remains. Here we demonstrate that solid solutions of the perovskites CaTaO2N and LaTaON2 constitute promising candidates for such substitutes: their brilliance, tinting strength, opacity, dispersability, light-fastness and heat stability rival that of the cadmium pigments, while their colour can be tuned through the desired range, from yellow through orange to deep red, by simple composition adjustments. Because all the constituent elements are harmless, this perovskite-based inorganic pigment system seems a promising replacement that could eliminate one of the sources for cadmium emissions to the environment and some of the remaining concerns about pigment safety. PMID- 10801124 TI - A homochiral metal-organic porous material for enantioselective separation and catalysis AB - Inorganic zeolites are used for many practical applications that exploit the microporosity intrinsic to their crystal structures. Organic analogues, which are assembled from modular organic building blocks linked through non-covalent interactions, are of interest for similar applications. These range from catalysis, separation and sensor technology to optoelectronics, with enantioselective separation and catalysis being especially important for the chemical and pharmaceutical industries. The modular construction of these analogues allows flexible and rational design, as both the architecture and chemical functionality of the micropores can, in principle, be precisely controlled. Porous organic solids with large voids and high framework stability have been produced, and investigations into the range of accessible pore functionalities have been initiated. For example, catalytically active organic zeolite analogues are known, as are chiral metal-organic open-framework materials. However, the latter are only available as racemic mixtures, or lack the degree of framework stability or void space that is required for practical applications. Here we report the synthesis of a homochiral metal-organic porous material that allows the enantioselective inclusion of metal complexes in its pores and catalyses a transesterification reaction in an enantioselective manner. Our synthesis strategy, which uses enantiopure metal-organic clusters as secondary building blocks, should be readily applicable to chemically modified cluster components and thus provide access to a wide range of porous organic materials suitable for enantioselective separation and catalysis. PMID- 10801125 TI - Recycled oceanic crust observed in 'ghost plagioclase' within the source of Mauna Loa lavas AB - The hypothesis that mantle plumes contain recycled oceanic crust is now widely accepted. Some specific source components of the Hawaiian plume have been inferred to represent recycled oceanic basalts, pelagic sediments or oceanic gabbros. Bulk lava compositions, however, retain the specific trace-element fingerprint of the original crustal component in only a highly attenuated form. Here we report the discovery of exotic, strontium-enriched melt inclusions in Mauna Loa olivines. Their complete trace-element patterns strongly resemble those of layered gabbros found in ophiolites, which are characterized by cumulus plagioclase with very high strontium abundances. The major-element compositions of these melts indicate that their composition cannot be the result of the assimilation of present-day oceanic crust through which the melts have travelled. Instead, the gabbro has been transformed into a (high-pressure) eclogite by subduction and recycling, and this eclogite has then been incorporated into the Hawaiian mantle plume. The trace-element signature of the original plagioclase is present only as a 'ghost' signature, which permits specific identification of the recycled rock type. The 'ghost plagioclase' trace-element signature demonstrates that the former gabbro can retain much of its original chemical identity through the convective cycle without completely mixing with other portions of the former oceanic crust. PMID- 10801126 TI - Human population in the biodiversity hotspots. AB - Biologists have identified 25 areas, called biodiversity hotspots, that are especially rich in endemic species and particularly threatened by human activities. The human population dynamics of these areas, however, are not well quantified. Here we report estimates of key demographic variables for each hotspot, and for three extensive tropical forest areas that are less immediately threatened. We estimate that in 1995 more than 1.1 billion people, nearly 20% of world population, were living within the hotspots, an area covering about 12% of Earth's terrestrial surface. We estimate that the population growth rate in the hotspots (1995-2000) is 1.8% yr(-1), substantially higher than the population growth rate of the world as a whole (1.3% yr(-1)) and above that of the developing countries (1.6% yr(-1)). These results suggest that substantial human induced environmental changes are likely to continue in the hotspots and that demographic change remains an important factor in global biodiversity conservation. The results also underline the potential conservation significance of the continuing worldwide declines in human fertility and of policies and programs that influence human migration. PMID- 10801127 TI - Identification of sleep-promoting neurons in vitro. AB - The neurons responsible for the onset of sleep are thought to be located in the preoptic area and more specifically, in the ventrolateral preoptic nucleus (VLPO). Here we identify sleep-promoting neurons in vitro and show that they represent an homogeneous population of cells that must be inhibited by systems of arousal during the waking state. We find that two-thirds of the VLPO neurons are multipolar triangular cells that show a low-threshold spike. This proportion matches that of cells active during sleep in the same region. We then show, using single-cell reverse transcriptase followed by polymerase chain reaction, that these neurons probably contain gamma-aminobutyric acid (GABA). We also show that these neurons are inhibited by noradrenaline and acetylcholine, both of which are transmitters of wakefulness. As most of these cells are also inhibited by serotonin but unaffected by histamine, their overall inhibition by transmitters of wakefulness is in agreement with their relative inactivity during waking with respect to sleep. We propose that the reciprocal inhibitory interaction of such VLPO neurons with the noradrenergic, serotoninergic and cholinergic waking systems to which they project is a key factor for promoting sleep. PMID- 10801128 TI - TACI and BCMA are receptors for a TNF homologue implicated in B-cell autoimmune disease. AB - B cells are important in the development of autoimmune disorders by mechanisms involving dysregulated polyclonal B-cell activation, production of pathogenic antibodies, and co-stimulation of autoreactive T cells. zTNF4 (BLyS, BAFF, TALL 1, THANK) is a member of the tumour necrosis factor (TNF) ligand family that is a potent co-activator of B cells in vitro and in vivo. Here we identify two receptors for zTNF4 and demonstrate a relationship between zTNF4 and autoimmune disease. Transgenic animals overexpressing zTNF4 in lymphoid cells develop symptoms characteristic of systemic lupus erythaematosus (SLE) and expand a rare population of splenic B-Ia lymphocytes. In addition, circulating zTNF4 is more abundant in NZBWF1 and MRL-lpr/lpr mice during the onset and progression of SLE. We have identified two TNF receptor family members, TACI and BCMA, that bind zTNF4. Treatment of NZBWF1 mice with soluble TACI-Ig fusion protein inhibits the development of proteinuria and prolongs survival of the animals. These findings demonstrate the involvement of zTNF4 and its receptors in the development of SLE and identify TACI-Ig as a promising treatment of autoimmune disease in humans. PMID- 10801129 TI - Transmembrane phosphoprotein Cbp regulates the activities of Src-family tyrosine kinases. AB - The Src family of protein tyrosine kinases (Src-PTKs) is important in the regulation of growth and differentiation of eukaryotic cells. The activity of Src PTKs in cells of different types is negatively controlled by Csk, which specifically phosphorylates a conserved regulatory tyrosine residue at the carboxy-terminal tail of the Src-PTKs. Csk is mainly cytoplasmic and Src-PTKs are predominantly membrane-associated. This raises a question about the mechanism of interaction between these enzymes. Here we present Cbp--a transmembrane phosphoprotein that is ubiquitously expressed and binds specifically to the SH2 domain of Csk. Cbp is involved in the membrane localization of Csk and in the Csk mediated inhibition of c-Src. In the plasma membrane Cbp is exclusively localized in the GM1 ganglioside-enriched detergent-insoluble membrane domain, which is important in receptor-mediated signalling. These findings reveal Cbp as a new component of the regulatory mechanism controlling the activity of membrane associated Src-PTKs. PMID- 10801130 TI - CpG methylation is maintained in human cancer cells lacking DNMT1. AB - Hypermethylation is associated with the silencing of tumour susceptibility genes in several forms of cancer; however, the mechanisms responsible for this aberrant methylation are poorly understood. The prototypic DNA methyltransferase, DNMT1, has been widely assumed to be responsible for most of the methylation of the human genome, including the abnormal methylation found in cancers. To test this hypothesis, we disrupted the DNMT1 gene through homologous recombination in human colorectal carcinoma cells. Here we show that cells lacking DNMT1 exhibited markedly decreased cellular DNA methyltransferase activity, but there was only a 20% decrease in overall genomic methylation. Although juxtacentromeric satellites became significantly demethylated, most of the loci that we analysed, including the tumour suppressor gene p16INK4a, remained fully methylated and silenced. These results indicate that DNMT1 has an unsuspected degree of regional specificity in human cells and that methylating activities other than DNMT1 can maintain the methylation of most of the genome. PMID- 10801131 TI - Direct observation of dendritic actin filament networks nucleated by Arp2/3 complex and WASP/Scar proteins. AB - Most nucleated cells crawl about by extending a pseudopod that is driven by the polymerization of actin filaments in the cytoplasm behind the leading edge of the plasma membrane. These actin filaments are linked into a network by Y-branches, with the pointed end of each filament attached to the side of another filament and the rapidly growing barbed end facing forward. Because Arp2/3 complex nucleates actin polymerization and links the pointed end to the side of another filament in vitro, a dendritic nucleation model has been proposed in which Arp2/3 complex initiates filaments from the sides of older filaments. Here we report, by using a light microscopy assay, many new features of the mechanism. Branching occurs during, rather than after, nucleation by Arp2/3 complex activated by the Wiskott-Aldrich syndrome protein (WASP) or Scar protein; capping protein and profilin act synergistically with Arp2/3 complex to favour branched nucleation; phosphate release from aged actin filaments favours dissociation of Arp2/3 complex from the pointed ends of filaments; and branches created by Arp2/3 complex are relatively rigid. These properties result in the automatic assembly of the branched actin network after activation by proteins of the WASP/Scar family and favour the selective disassembly of proximal regions of the network. PMID- 10801132 TI - Low fidelity DNA synthesis by human DNA polymerase-eta. AB - A superfamily of DNA polymerases that bypass lesions in DNA has been described. Some family members are described as error-prone because mutations that inactivate the polymerase reduce damage-induced mutagenesis. In contrast, mutations in the skin cancer susceptibility gene XPV, which encodes DNA polymerase (pol)-eta, lead to increased ultraviolet-induced mutagenesis. This, and the fact that pol-eta primarily inserts adenines during efficient bypass of thymine-thymine dimers in vitro, has led to the description of pol-eta as error free. However, here we show that human pol-eta copies undamaged DNA with much lower fidelity than any other template-dependent DNA polymerase studied. Pol-eta lacks an intrinsic proofreading exonuclease activity and, depending on the mismatch, makes one base substitution error for every 18 to 380 nucleotides synthesized. This very low fidelity indicates a relaxed requirement for correct base pairing geometry and indicates that the function of pol-eta may be tightly controlled to prevent potentially mutagenic DNA synthesis. PMID- 10801133 TI - Roles of E. coli DNA polymerases IV and V in lesion-targeted and untargeted SOS mutagenesis. AB - The expression of the Escherichia coli DNA polymerases pol V (UmuD'2C complex) and pol IV (DinB) increases in response to DNA damage. The induction of pol V is accompanied by a substantial increase in mutations targeted at DNA template lesions in a process called SOS-induced error-prone repair. Here we show that the common DNA template lesions, TT (6-4) photoproducts, TT cis-syn photodimers and abasic sites, are efficiently bypassed within 30 seconds by pol V in the presence of activated RecA protein (RecA*), single-stranded binding protein (SSB) and pol III's processivity beta,gamma-complex. There is no detectable bypass by either pol IV or pol III on this time scale. A mutagenic 'signature' for pol V is its incorporation of guanine opposite the 3'-thymine of a TT (6-4) photoproduct, in agreement with mutational spectra. In contrast, pol III and pol IV incorporate adenine almost exclusively. When copying undamaged DNA, pol V exhibits low fidelity with error rates of around 10(-3) to 10(-4), with pol IV being 5- to 10 fold more accurate. The effects of RecA protein on pol V, and beta,gamma-complex on pol IV, cause a 15,000- and 3,000-fold increase in DNA synthesis efficiency, respectively. However, both polymerases exhibit low processivity, adding 6 to 8 nucleotides before dissociating. Lesion bypass by pol V does not require beta,gamma-complex in the presence of non-hydrolysable ATPgammaS, indicating that an intact RecA filament may be required for translesion synthesis. PMID- 10801134 TI - Retrotransposition of a bacterial group II intron. AB - Self-splicing group II introns may be the evolutionary progenitors of eukaryotic spliceosomal introns, but the route by which they invade new chromosomal sites is unknown. To address the mechanism by which group II introns are disseminated, we have studied the bacterial L1.LtrB intron from Lactococcus lactis. The protein product of this intron, LtrA, possesses maturase, reverse transcriptase and endonuclease enzymatic activities. Together with the intron, LtrA forms a ribonucleoprotein (RNP) complex which mediates a process known as retrohoming. In retrohoming, the intron reverse splices into a cognate intronless DNA site. Integration of a DNA copy of the intron is recombinase independent but requires all three activities of LtrA. Here we report the first experimental demonstration of a group II intron invading ectopic chromosomal sites, which occurs by a distinct retrotransposition mechanism. This retrotransposition process is endonuclease-independent and recombinase-dependent, and is likely to involve reverse splicing of the intron RNA into cellular RNA targets. These retrotranspositions suggest a mechanism by which splicesomal introns may have become widely dispersed. PMID- 10801135 TI - From bench to bedside in cardiovascular research a reflection on my research career. PMID- 10801136 TI - A novel feature disclosed in opticospinal multiple sclerosis in Asians. PMID- 10801137 TI - Gastric mucosa-associated lymphoid tissue lymphoma and Helicobacter pylori infection: evaluation of antibiotic treatment. PMID- 10801138 TI - Intestinal necrosis associated with severe acute pancreatitis. PMID- 10801139 TI - Pulmonary diseases due to Mycobacterium szulgai in Japan. PMID- 10801140 TI - Diagnosis and treatment of pulmonary hypertension associated with collagen diseases. PMID- 10801141 TI - Juvenile muscular atrophy of distal upper extremity (Hirayama disease). AB - This disease is characterized by initially progressive muscular weakness and wasting of the distal upper limb(s) in young people predominantly in men, followed by a spontaneous arrest within several years. This disease has been thought to be separate from motor neuron diseases, yet some authors still consider the illness a variant of motor neuron disease. However, the pathological evidence of ischemic changes in the lower cervical anterior horn should facilitate differentiation of the disorder from degenerative motor neuron disease. Recent radiological investigations proved compressive flattening of the lower cervical cord due to forward displacement of the cervical dural sac and spinal cord induced by neck flexion. These findings suggest that sustained or repeated neck flexion may cause ischemic changes in the cervical anterior horn. Application of a cervical collar to minimize neck flexion prevents progressive muscular weakness in an early stage of the disease. PMID- 10801142 TI - Changes in squamous cell carcinoma-related antigen levels before and after hemodialysis in relation to the model of dialyzer employed. AB - OBJECTIVE: Although tumor marker levels in blood become elevated after hemodialysis as a result of the amount of fluid removed, serum squamous cell carcinoma-related antigen (SCC) levels have not been always reported to increase after hemodialysis. The purpose of this report is to determine whether there is a difference between the change of serum SCC levels before and after hemodialysis according to the model of dialyzer employed. PATIENTS AND METHODS: In ninety-four patients on hemodialysis (50 cases of diabetic nephropathy and 44 cases of glomerulonephritis), we examined serum SCC levels before and after hemodialysis. RESULTS: There was no overall difference between SCC levels before and after hemodialysis (3.2+/-1.5 ng/ml vs 3.3+/-1.7 ng/ml, p=0.2381). In patients treated with a cellulosic type membrane dialyzer (n=73), SCC levels after hemodialysis were higher than those before hemodialysis (3.7+/-1.7 ng/ml vs 3.5+/-1.5 ng/ml, p=0.0495). In patients treated with a synthetic type membrane (n=21), SCC levels after hemodialysis decreased when compared to those before hemodialysis (1.9+/ 0.7 ng/ml vs 2.2+/-0.8 ng/ml, p=0.0018) and in all patients, the SCC levels after hemodialysis were lower than, or equal to, those before hemodialysis. CONCLUSION: The results suggest that the decline in serum SCC levels during hemodialysis treated with synthetic type membrane of dialyzers, concealed the increased SCC levels in hemoconcentration. PMID- 10801143 TI - Hyperprolactinemia in optico-spinal multiple sclerosis. AB - OBJECTIVE: To clarify the clinical features of MS patients with hyperprolactinemia. SUBJECTS AND METHODS: The serum prolactin level was measured in 67 Japanese patients (19 men and 48 women) with multiple sclerosis (MS) and in 16 patients (4 men and 12 women) with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) using a two-site immunoradiometric assay. RESULTS: In the MS patients, 32 were classified as having Asian type MS showing a selective involvement of the optic nerves and spinal cord, while the other 35 were classified as having Western type MS which displayed disseminated central nervous system involvement. In women, the serum prolactin level was found to be significantly higher only in Asian type MS (mean=23.1 ng/ml, n=25) than in HAM/TSP (mean=6.9 ng/ml, n=12) (p=0.0297), while it did not differ significantly in men among the three groups. Hyperprolactinemia was significantly associated with acute relapse involving the optic nerves. All MS patients with hyperprolactinemia (7 women with Asian type MS and 2 women with Western type MS) showed recurrent opticomyelitis either throughout or in the early course of the disease, and also had a higher age of onset, a higher Expanded Disability Status Scale score, a greater visual impairment, and higher cell counts and protein contents in the cerebrospinal fluid than did the normoprolactinemic patients. CONCLUSION: Hyperprolactinemia may be one of the characteristic features of Asian patients with MS who preferentially show the optic nerve involvement. PMID- 10801144 TI - Regression of gastric mucosa-associated lymphoid tissue lymphoma with reduced telomerase activity after eradication of Helicobacter pylori. AB - Recently, telomerase activity has been demonstrated in a large number of malignant tumors whereas its activity is not detected in most normal somatic cells suggesting its role in the immortalization process. Here we report the first investigation of telomerase activity in a case of gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Elevated telomerase activity was detected in biopsy specimens of the lymphoma. After eradication of Helicobacter pylori, the level of telomerase activity returned to normal with histological regression of the lymphoma. The telomerase activity was associated with the disease activity of the gastric MALT lymphoma after eradication therapy in the present case. PMID- 10801145 TI - Development of colonic necrosis following severe acute pancreatitis. AB - We herein describe a 70-year-old male patient who developed colonic necrosis following severe acute pancreatitis. He was referred to our hospital with a diagnosis of acute pancreatitis. In the course of the disease, he developed sudden and massive hematochezia and died. The autopsy findings revealed large bowel ischemia with transmural infarction. The possible pathogenic mechanisms of colonic ischemia are also discussed. PMID- 10801146 TI - Pulmonary mycobacteriosis caused by rifampicin-resistant Mycobacterium szulgai. AB - We describe a rare case of pulmonary mycobacteriosis infected with rifampicin (RFP)-resistant Mycobacterium szulgai that was successfully eradicated with clarithromycin (CAM) treatment. An 80-year-old man was admitted to our hospital with a 4-week history of high fever, productive cough and malaise. Chest roentgenogram showed an infiltrative shadow in the left lower lung field. Isolated bacteria from sputum were acid-fast bacilli and identified as M. szulgai by the DNA-DNA hybridization method. Drug susceptibility tests showed resistance to RFP (MIC>100 microg/ml). Combined treatment with ethionamide, streptomycin and isoniazid based on the results of drug susceptibility tests resulted in clinical and radiologic improvement within two years. Additional treatment with CAM for another year resulted in complete eradication of the mycobacterium. PMID- 10801147 TI - Magnetic resonance imaging findings in primary amyloidosis-associated arthropathy. AB - The MRI findings of amyloid arthropathy associated with primary amyloidosis are presented here possibly for the first time in the literature. Two types of lesions are noted: (1) capsular and tendon lesions; these regions are thickened, hypointense and enhanced by gadolinium (Gd) on T1 weighted imaging (T1WI), and hyperintense on T2 weighted imaging (T2WI), and (2) periarticular and osseous lesions; these regions appear to be tumor-forming and hypointense on both T1WI and T2WI and are not enhanced by Gd. It is necessary to differentiate these findings from other diseases such as chondrosarcoma, rhabdomyosarcoma and chronic inflammatory lesions such as tuberculosis. PMID- 10801148 TI - Successful treatment of systemic lupus erythematosus and pulmonary hypertension with intravenous prostaglandin I2 followed by its oral analogue. AB - Systemic lupus erythematosus (SLE) is sometimes reported to complicate fatal pulmonary hypertension. A 46-year-old woman, with a ten-year history of SLE and pulmonary hypertension, was admitted to our hospital complaining of dyspnea and chest pain. She suffered pulmonary hemorrhage and after steroid pulse therapy, she underwent continuous intravenous infusion of epoprostenol (prostaglandin I2) with corticosteroid for four weeks, which reduced the pulmonary artery pressure and resistance. Following the successful treatment, beraprost sodium, an oral PGI2 analogue, was given and it maintained pulmonary hypertension remittance for four years. PMID- 10801150 TI - The genes responsible for human progeroid syndromes. PMID- 10801149 TI - Oxidative stress and aging. PMID- 10801151 TI - Neuronal death in Alzheimer's disease. PMID- 10801152 TI - Development of acute coronary syndrome and progression of coronary artery disease: a serial clinical-angiographic analysis. PMID- 10801153 TI - The role of plaque rupture in the development of acute coronary syndrome evaluated by the coronary angioscope. PMID- 10801154 TI - Plaque formation and its rupture. PMID- 10801155 TI - Percutaneous coronary intervention in acute coronary syndrome. PMID- 10801156 TI - Current topics in the treatment of peptic ulcer. PMID- 10801157 TI - Recent advances in therapy for patients with inflammatory bowel disease. PMID- 10801158 TI - Autoimmune liver disease: current therapy. PMID- 10801159 TI - Troglitazone in progesterone derivative-induced impairment of glucose tolerance. PMID- 10801160 TI - Measurement of serum concentrations of cardiac troponin T in patients with hypereosinophilic syndrome: a sensitive non-invasive marker of cardiac disorder. PMID- 10801161 TI - Politicisation of debate on HIV care in South Africa. PMID- 10801162 TI - Choice of hormonal therapy for prostate cancer. PMID- 10801163 TI - Lessons about antibodies in autoimmune hepatitis. PMID- 10801164 TI - Pathogenesis and treatment of acute chest syndrome of sickle-cell anaemia. PMID- 10801165 TI - Surgery for primary hyperparathyroidism--sooner rather than later. PMID- 10801166 TI - Host factors that affect outcome of total hip arthroplasty. PMID- 10801167 TI - Antibiotics without prescription: "bacterial or medical resistance"? PMID- 10801168 TI - Novel oral treatment of Gaucher's disease with N-butyldeoxynojirimycin (OGT 918) to decrease substrate biosynthesis. AB - BACKGROUND: Current treatment for Gaucher's disease involves administration of intravenous glucocerebrosidase to degrade glucocerebroside stored in lysosomes. Lowering the rate of biosynthesis of glucocerebroside should decrease accumulation of this substrate. We investigated the safety and efficacy of OGT 918 (N-butyldeoxynojirimycin), an inhibitor of glucosyltransferase, as a novel oral treatment for non-neuronopathic Gaucher's disease. METHODS: We recruited, into a 1-year open-label study, 28 adults (seven with previous splenectomies) from four national Gaucher's referral clinics, who were unable or unwilling to receive enzyme treatment. We measured liver and spleen volume by computed tomography or magnetic resonance imaging at baseline and at months 6 and 12, and biochemical and haematological variables monthly, including chitotriosidase activity (a sensitive marker of Gaucher's disease activity). Patients were started on 100 mg oral OGT 918 three times daily. FINDINGS: Baseline liver volumes were 1.1-2.7 times normal and spleen volumes 5.1-24.8 times normal. At 12 months, mean liver and spleen volumes were significantly lowered by 12% (95% CI 7.8-16.4) and 19% (14.3-23.7), respectively (each p<0.001). Haematological variables improved slightly. Mean organ volume and blood counts improved continually between 6 months and 12 months of treatment. Mean chitotriosidase concentrations fell by 16.4% over 12 months (p<.0001). Six patients withdrew because of gastrointestinal complaints (two), personal reasons (two), or severe pre-existing disease (two). The most frequent adverse effect was diarrhoea, which occurred in 79% of patients shortly after the start of treatment. INTERPRETATION: Decrease of substrate formation by OGT 918 improves key clinical features of non neuronopathic Gaucher's disease. The strategy justifies further trials in this and other glycosphingolipid storage disorders. PMID- 10801169 TI - Adjunctive non-pharmacological analgesia for invasive medical procedures: a randomised trial. AB - BACKGROUND: Non-pharmacological behavioural adjuncts have been suggested as efficient safe means in reducing discomfort and adverse effects during medical procedures. We tested this assumption for patients undergoing percutaneous vascular and renal procedures in a prospective, randomised, single-centre study. METHODS: 241 patients were randomised to receive intraoperatively standard care (n=79), structured attention (n=80), or self-hypnotic relaxation (n=82). All had access to patient-controlled intravenous analgesia with fentanyl and midazolam. Patients rated their pain and anxiety on 0-10 scales before, every 15 min during and after the procedures. FINDINGS: Pain increased linearly with procedure time in the standard group (slope 0.09 in pain score/15 min, p<0.0001), and the attention group (slope 0.04/15 min; p=0.0425), but remained flat in the hypnosis group. Anxiety decreased over time in all three groups with slopes of -0.04 (standard), -0.07 (attention), and -0.11 (hypnosis). Drug use in the standard group (1.9 units) was significantly higher than in the attention and hypnosis groups (0.8 and 0.9 units, respectively). One hypnosis patient became haemodynamically unstable compared with ten attention patients (p=0.0041), and 12 standard patients (p=0.0009). Procedure times were significantly shorter in the hypnosis group (61 min) than in the standard group (78 min, p=0.0016) with procedure duration of the attention group in between (67 min). INTERPRETATION: Structured attention and self-hypnotic relaxation proved beneficial during invasive medical procedures. Hypnosis had more pronounced effects on pain and anxiety reduction, and is superior, in that it also improves haemodynamic stability. PMID- 10801170 TI - Maximum androgen blockade in advanced prostate cancer: an overview of the randomised trials. Prostate Cancer Trialists' Collaborative Group. AB - BACKGROUND: In advanced prostate cancer, androgen suppression (AS) by surgery or drugs controls testicular hormone secretion, and the further addition of an antiandrogen such as nilutamide, flutamide, or cyproterone acetate is referred to as maximum androgen blockade (MAB). The aim of this overview was to compare the effects on the duration of survival of MAB and of AS alone. METHODS: The collaborative meta-analysis of 27 randomised trials involved central reanalysis of the data on each of 8275 men (98% of those ever randomised in trials of MAB vs AS) with metastatic (88%) or locally advanced (12%) prostate cancer. Half were over 70 years of age, and follow-up was typically for about 5 years. FINDINGS: 5932 (72%) men have died; of the deaths for which causes were provided, about 80% were attributed to prostate cancer. 5-year survival was 25.4% with MAB versus 23.6% with AS alone, a non-significant gain of 1.8% (SE 1.3; logrank 2p=0.11). There was no significant heterogeneity in the treatment effect (MAB vs AS) with respect to age or disease stage. The results for cyproterone acetate, which accounted for only a fifth of the evidence, appeared slightly unfavourable to MAB (5-year survival 15.4% MAB vs 18.1% AS alone; difference -2.8% [SE 2.4]; logrank 2p=0.04 adverse), whereas those for nilutamide and flutamide appeared slightly favourable (5-year survival 27.6% MAB vs 24.7% AS alone; difference 2.9% [SE 1.3]; logrank 2p=0.005). Non-prostate-cancer deaths (although not clearly significantly affected by treatment) accounted for some of the apparently adverse effects of cyproterone acetate. INTERPRETATION: In advanced prostate cancer, addition of an antiandrogen to AS improved the 5-year survival by about 2% or 3% (depending on whether the analysis includes or excludes the cyproterone acetate trials), but the range of uncertainty as to the true size of this benefit runs from about 0% to about 5%. PMID- 10801171 TI - Poor bone quality or hip structure as risk factors affecting survival of total hip arthroplasty. AB - BACKGROUND: The principal long-term complication after total hip arthroplasty (THA) has been aseptic fixation failure. Many hip prostheses and operative techniques have been developed to improve outcomes, but few measures have been taken to cope with poor bone quality or hip structure. We assessed risk factors for aseptic fixation failure after THA. METHODS: We assessed, by multivariate analysis, survival of 405 primary Charnley THAs to identify risk factors for aseptic fixation failures. We also investigated risk factors for development of rapid polyethylene wear (penetration depth of the femoral head into the socket polyethylene > or = 2 mm/year) FINDINGS: In the entire series of 405 THAs, with use of radiographic fixation failure or revision for a loose socket as the endpoint, development of rapid polyethylene wear and the preoperative diagnosis of atrophic osteoarthrosis (defined by scarce osteophyte formation) were identified as risk factors for socket loosening (p < or = 0.02). A medullary canal with an unfavourable geometry (a stovepipe canal, Noble's canal-flare index < 3.0) was the only risk factor for femoral fixation failure (p < or = 6.7x10(3)). The only variable related to development of rapid polyethylene wear was the type of steel used in the femoral prosthesis--Ortron 90 prostheses significantly lowered the rate of development of rapid wear from 12.7% to 0.4%. In the 248 THAs in which these femoral prostheses were used, socket survival was affected only by the preoperative diagnosis of atrophic osteoarthrosis (for radiographic fixation failure and revision, p=4.0x10(-5) and p=0.042, respectively). INTERPRETATION: In THA, the critical risk factors are poor bone quality, which manifests as atrophic osteoarthrosis, for socket survival and poor bone structure for femoral-prosthesis survival. To ensure longer durability of THAs, these factors should be assessed further and efforts, especially biological initiatives, should be made to resolve them. PMID- 10801172 TI - Radiotherapy for Graves' orbitopathy: randomised placebo-controlled study. AB - BACKGROUND: The best treatment (steroids, irradiation, or both) for moderately severe Graves' orbitopathy, a self-limiting disease is not known. We tested the efficacy of external beam irradiation compared with sham-irradiation. METHODS: In a double-blind randomised clinical trial, 30 patients with moderately severe Graves' orbitopathy had radiotherapy (20 Gy in ten fractions), and 30 were assigned sham-irradiation (ten fractions of 0 Gy). Treatment outcome was measured qualitatively by changes in major and minor criteria and quantitatively in several ophthalmic and other variables, such as eyelid aperture, proptosis, eye movements, subjective eye score, and clinical-activity score at 24 weeks. FINDINGS: The qualitative treatment outcome was successful in 18 of 30 (60%) irradiated patients versus nine of 29 (31%) sham-irradiated patients at week 24 (relative risk [RR]=1.9 [95% CI 1.0-3.6], p=0.04). This difference was caused by improvements in diplopia grade, but not by reduction of proptosis, nor of eyelid swelling. Quantitatively, elevation improved significantly in the radiotherapy group, whereas all other variables remained unchanged. The field of binocular single vision was enlarged in 11 of 17 patients after irradiation compared with two of 15 after sham-irradiation. Nevertheless, only 25% of the irradiated patients were spared from additional strabismus surgery. INTERPRETATION: In these patients with moderately severe Graves' orbitopathy, radiotherapy should be used only to treat motility impairment. PMID- 10801174 TI - A man with chronic facial swelling. PMID- 10801173 TI - Identification of target antigen for SLA/LP autoantibodies in autoimmune hepatitis. AB - BACKGROUND: Autoantibodies are a hallmark of autoimmune hepatitis, but most are not disease specific. Autoantibodies to soluble liver antigen (SLA) and to liver and pancreas antigen (LP) have been described as disease specific, occurring in about 30% of all patients with autoimmune hepatitis, but no standardised assays are available. Methods We tested 2000 serum samples from patients with various liver diseases and controls for SLA autoantibodies by inhibition ELISA. Serum samples positive for SLA antibodies were used for immunoscreening of cDNA expression libraries. Identified clones were tested against a panel of serum samples positive for SLA and LP autoantibodies and control serum samples, and the epitope mapped by deletion mutants and exonuclease digestion. FINDINGS: SLA and LP autoantibodies were identical. Of 2000 serum samples screened, 35 were positive for SLA autoantibodies. These positive samples came from patients with autoimmune hepatitis; three from patients with an overlap syndrome (primary biliary cirrhosis and secondary autoimmune hepatitis). Expression cloning and absorption experiments identified a 422 aminoacid protein present in two splice variants as the sole target antigen. Aminoacids 371-409 were critical for immune recognition. INTERPRETATION: The identified cDNA encodes the primary target antigen of SLA/LP autoantibodies. The SLA/LP antigen has a previously unknown aminoacid sequence, and presumably codes for an unindentified enzyme, suggested to be UGA-suppressor tRNA-associated protein. SLA/LP autoantibodies are disease specific and recognise a dominant epitope, suggesting a specific antigen-driven immune response. Identification of the SLA/LP target antigen will allow establishment of a reliable, widely available diagnostic assay. Furthermore, its role in the pathogenesis of autoimmune hepatitis can now be studied. PMID- 10801175 TI - Relation between airborne pollen concentrations and daily cardiovascular and respiratory-disease mortality. AB - In a time-series study in The Netherlands, we found a strong association between the day-to-day variation in pollen concentrations and that of deaths due to cardiovascular disease, chronic obstructive pulmonary disease, and pneumonia. PMID- 10801176 TI - Zonulin, a newly discovered modulator of intestinal permeability, and its expression in coeliac disease. AB - We identified zonulin, a novel human protein analogue to the Vibrio cholerae derived Zonula occludens toxin, which induces tight junction disassembly and a subsequent increase in intestinal permeability in non-human primate intestinal epithelia. Zonulin expression was raised in intestinal tissues during the acute phase of coeliac disease, a clinical condition in which tight junctions are opened and permeability is increased. PMID- 10801177 TI - Detection of human herpes virus 6 DNA in fetal hydrops. AB - Human herpes virus 6 (HHV6) DNA was detected in two of eight fetuses with hydrops and none of ten non-hydropic dead fetuses. Both cases with HHV6 DNA had chromosomal abnormalities. Positive results were confirmed with a second PCR specific for an alternate region of the HHV6 genome. Restriction endonuclease analysis confirmed that the viral DNA was representative of HHV6 type A. PMID- 10801178 TI - Amplification of DNA sequences in polar bodies from human oocytes for diagnosis of mitochondrial disease. AB - When a preovulatory oocyte reinitiates meiosis, it sheds a cytoplasmic fragment containing mitochondria and a redundant set of chromosomes. We have detected DNA sequences from both the mitochondrial and nuclear genomes in polar bodies from unfertilised human oocytes, demonstrating the feasibility of diagnosing mitochondrial diseases before conception. PMID- 10801179 TI - Effect of consumption of red wine, spirits, and beer on serum homocysteine. AB - Serum homocysteine increases after moderate consumption of red wine and spirits, not after moderate consumption of beer. Vitamin B6 in beer seems to prevent the alcohol-induced rise in serum homocysteine. PMID- 10801180 TI - FDA approves first drug in new class of antibiotics. PMID- 10801181 TI - New US tuberculosis guidelines suggest targeted screening. PMID- 10801182 TI - Genetic testing and insurance: fears unfounded? PMID- 10801183 TI - Slow progress towards protecting people in US clinical trials. PMID- 10801185 TI - Remote Indian villages to benefit from telemedicine project. PMID- 10801184 TI - WHO programme gives hope to blind and partially sighted people in Africa. PMID- 10801186 TI - Platelets. AB - Platelets, derived from megakaryocyte cytoplasm, have a critical role in normal haemostasis, and in thrombotic disorders. The development of megakaryocytes is controlled by thrombopoietin, which binds to c-mpl on the surface of platelets and megakaryocytes. Platelet membrane glycoproteins mediate binding to subendothelial tissue and aggregation into haemostatic plugs. Thrombocytopenia and disorders of platelet function cause petechiae and mucocutaneous bleeding. Drugs causing specific inhibition of platelet function are important in the treatment of cardiovascular and cerebrovascular disease. PMID- 10801187 TI - Database of patients' experiences (DIPEx): a multi-media approach to sharing experiences and information. AB - The Database of Individual Patients' Experience of illness (DIPEx) is a multimedia website and CD-ROM that links patients' experiences with evidence based information about treatments and the illness itself and with a range of other resources that may be useful, including support groups and links to other websites. DIPEx aims to identify the questions that matter to people when they are ill and it has potential for informing patients, educating healthcare professionals, and providing a patient-centred perspective to researchers and those who manage health services. PMID- 10801188 TI - Nuclear weapons, a continuing threat to health. AB - 32,000 nuclear weapons, with a destructive force equivalent to several thousand megatons of conventional explosive, are still deployed. The risk of nuclear war by accident may have increased and new threats include war between newly declared nuclear-weapon-states and the construction by terrorist groups of crude but effective devices. Health workers have drawn attention in the past to the likely major health consequences of the use of nuclear weapons. An opportunity for their global elimination under a nuclear weapons convention arises with the current review conference in New York of the nuclear Non-Proliferation Treaty--a crucial event for efforts to bring about a world free of nuclear weapons. PMID- 10801189 TI - The rights of the severely mentally ill in post-conflict societies. PMID- 10801190 TI - Tools to measure landmine incidents and injuries. PMID- 10801191 TI - Torture and the scientific community. PMID- 10801192 TI - Long-term outcome of invasive breast cancer. PMID- 10801193 TI - Long-term outcome of invasive breast cancer. PMID- 10801194 TI - Long-term outcome of invasive breast cancer. PMID- 10801195 TI - Long-term outcome of invasive breast cancer. PMID- 10801196 TI - Pertussis transmission in England and Wales. PMID- 10801197 TI - Toxicity of old and new antidepressant drugs. PMID- 10801198 TI - Another look at latex. PMID- 10801199 TI - High-risk behaviour. PMID- 10801200 TI - High-risk behaviour. PMID- 10801201 TI - Post-exposure prophylaxis for HIV infection. Italian Registry of Post-Exposure Prophylaxis. PMID- 10801202 TI - Chimeric and humanised--misunderstood. PMID- 10801203 TI - Measles genotype G2 in Indonesia and Malaysia. PMID- 10801204 TI - Discontinuation of doxazosin arm of ALLHAT. Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack. PMID- 10801205 TI - Encephalopathy associated with influenza epidemics. PMID- 10801206 TI - Black people's red faces and AIDS prevention. PMID- 10801207 TI - Forecasting disease risk with seasonal climate predictions. PMID- 10801208 TI - Pliny, Galileo--and the late Bill Hamilton. PMID- 10801209 TI - A shocking case of diabetic neuropathy. PMID- 10801210 TI - Effects of transmit focusing on finite amplitude distortion based second harmonic generation. AB - Generation of the second harmonic signal was studied for finite amplitude distortion based harmonic imaging. Acoustic field amplitudes along the range axis of a fixed focus transducer were measured using a PVDF needle hydrophone. Results indicated that on-axis amplitudes strongly depended on the f-number at both the fundamental and the second harmonic frequencies. Differences of the on-axis amplitudes between the two frequencies were also investigated. To explore the possibility of increasing harmonic generation by extending the depth of focus, a two-focus transducer was employed. Hydrophone measurements, pulse-echo imaging and simulations were performed. Although the increase in harmonic generation depended on specific imaging parameters, the effectiveness of improving the harmonic signal-to-noise ratio (SNR) by increasing the depth of field was clearly demonstrated. Degradation in contrast resolution associated with the two-focus transducer was also evaluated. It was found that the contrast resolution was still significantly better than that of the fundamental image at the same frequency. Results of the study using the two-focus transducers can be generalized to imaging systems with full dynamic transmit focusing capabilities. In other words, it is expected that dynamic transmit focusing can improve the SNR of finite amplitude distortion based second harmonic imaging while improving the contrast resolution over fundamental imaging. PMID- 10801211 TI - Time-resolved pulsed elastography with ultrafast ultrasonic imaging. AB - In this paper, a new elastographic method is proposed. Using this method, the propagation of a low-frequency transient shear wave can be imaged by means of an ultrafast imaging system (up to 10,000 frames/s) that we have developed. Ultrafast ultrasonic imaging is obtained with a linear array of transducers (3.5 MHz) connected to electronics that have 64 channels sampled at 30 MHz and 128 Kbytes for storing the backscattered signals. Displacements are measured using cross-correlation of the ultrasonic signals. Movies of the low central frequency (200 Hz) shear wave propagation through homogeneous and heterogeneous phantoms have been obtained with 1,000 and 2,000 frames per second. PMID- 10801212 TI - Clinical trial simulation in drug development. AB - Clinical trial simulation is the application of old technologies, e.g., Monte Carlo simulation, to a new problem, that problem being how to maximize the information content obtained during the drug development process with an intent to have the greatest chance of "success" in a clinical trial. When the information content of the drug is high, then simulation provides a method to synthesize that information into a coherent package that indicates the sponsor has good control over the pharmacology of the drug. From a purely financial point of view, what simulation offers pharmaceutical companies is the possibility of reducing the number of required studies, maximizing the chances for success in a clinical trial, and possibly shortening development time; all outcomes which will reduce drug development costs. The purpose of this paper is to introduce clinical trial simulation to the reader by discussing its potential in drug development, to briefly review the literature, and to make recommendations and caveats regarding its use. PMID- 10801213 TI - Principles and application of ultrasound in pharmaceutical powder compression. AB - The use of ultrasound during the tableting of pharmaceutical powders is a new concept. However, in the metallurgy, plastic, and ceramic industries, ultrasound assisted compression of materials has been known for some years. Ultrasound improves the characteristics of the compression process leading to optimized mechanical strength of the compacts without applying excessive compression force. Therefore, problems associated with high-pressure compression in tableting can be overcome and tablets may be manufactured more economically and consistently with the aid of ultrasound compared to conventional pressure processes. Although great progress in the theoretical understanding of the ultrasound-assisted powder compression process has been made since the late 1960s, the need for further research in the area of ultrasound application during pharmaceutical powder compression is essential. Further investigations on a wide range of drugs and excipients, to expand the usefulness and scope of the ultrasound-assisted technique, and to understand the complex phenomena involved in the process, are needed. In this article the principles, advantages, and limitations of the application of ultrasonic vibrations during pharmaceutical powder compression is reviewed with the hope that this article can contribute to, and stimulate research in the area. PMID- 10801214 TI - Transport across the primate blood-brain barrier of a genetically engineered chimeric monoclonal antibody to the human insulin receptor. AB - PURPOSE: Brain drug targeting may be achieved by conjugating drugs, that normally do not cross the blood-brain barrier (BBB), to brain drug delivery vectors. The murine 83-14 MAb to the human insulin receptor (HIR) is a potential brain drug targeting vector that could be used in humans, if this MAb was genetically engineered to form a chimeric antibody. where most of the immunogenic murine sequences are replaced by human antibody sequence. METHODS: The present studies describe the production of the gene for the chimeric HIRMAb, expression and characterization of the protein, radiolabeling of the chimeric HIRMAb with 111 indium and 125-iodine, and quantitative autoradiography of living primate brain taken 2 hours after intravenous administration of the [111In]chimeric HIRMAb. RESULTS: The chimeric HIRMAb had identical affinity to the target antigen as the murine HIRMAb based on Western blotting and immunoradiometric assay using partially purified HIR affinity purified from serum free conditioned media produced by a CHO cell line secreting soluble HIR. The [125I]chimeric HIRMAb was avidly bound to isolated human brain capillaries, and this binding was blocked by the murine HIRMAb. The [111In]chimeric HIRMAb was administered intravenously to an anesthetized Rhesus monkey, and the 2 hour brain scan showed robust uptake of the chimeric antibody by the living primate brain. CONCLUSIONS: A genetically engineered chimeric HIRMAb has been produced, and the chimeric antibody has identical reactivity to the human and primate BBB HIR as the original murine antibody. This chimeric HIRMAb may be used in humans for drug targeting through the BBB of neurodiagnostic or neurotherapeutic drugs that normally do not cross the BBB. PMID- 10801215 TI - Novel lipid-based hollow-porous microparticles as a platform for immunoglobulin delivery to the respiratory tract. AB - PURPOSE: Delivery of specific antibodies or immunoglobulin constructs to the respiratory tract may be useful for prophylaxis or active treatment of local or systemic disorders. Therefore, we evaluated the utility of lipid-based hollow porous microparticles (PulmoSpheres) as a potential delivery vehicle for immunoglobulins. METHODS: Lipid-based microparticles loaded with human immunoglobulin (hIgG) or control peptide were synthesized by spray drying and tested for: i) the kinetics of peptide/protein release, using ELISA and bioassays; ii) bioavailability subsequent to nonaqueous liquid instillation into the respiratory tract of BALB/c mice, using ELISA and Western blotting; iii) bioactivity in terms of murine immune response to xenotypic epitopes on human IgG, using ELISA and T cell assays; and iv) mechanisms responsible for the observed enhancement of immune responses, using measurement of antibodies as well as tagged probes. RESULTS: Human IgG and the control peptide were both readily released from the hollow-porous microspheres once added to an aqueous environment, although the kinetics depended on the compound. Nonaqueous liquid instillation of hIgG formulated in PulmoSpheres into the upper and lower respiratory tract of BALB/c mice resulted in systemic biodistribution. The formulated human IgG triggered enhanced local and systemic immune responses against xenotypic epitopes and was associated with receptor-mediated loading of alveolar macrophages. CONCLUSIONS: Formulation of immunoglobulins in hollow porous microparticles is compatible with local and systemic delivery via the respiratory mucosa and may be used as means to trigger or modulate immune responses. PMID- 10801216 TI - Regional differences in intestinal spreading and pH recovery and the impact on salmon calcitonin absorption in dogs. AB - PURPOSE: To investigate the regional influence of intestinal spreading and pH recovery on the performance of drug and excipient delivery systems and their impact on the oral absorption of a model peptide drug, salmon calcitonin (sCT), in conscious beagle dogs. METHODS: Male beagle dogs were surgically prepared with subdermal Intestinal Access Ports (IAP). The catheter from one port was placed in the duodenum and the other in the ileum. Fluoroscopy and Heidelberg pH capsule studies were performed to characterize intestinal spreading and pH recovery, respectively. Three treatments were performed: (1) a radiopaque dye and citric acid (CA) were infused into the intestinal segments, (2) a radiopaque powder capsule containing CA was given orally, and (3) capsules containing CA and sCT were given orally. Regular blood samples were collected and analyzed by radioimmunoassay (RIA) to determine the absorption characteristics of sCT. RESULTS: Since sCT is an excellent substrate for the pancreatic serine protease trypsin, the rate of degradation of sCT in the GI lumen is dependent upon the regional pH, activity of digestive enzymes and the concentration of sCT at the site of absorption. Fluoroscopy results clearly showed that when the radiopaque dye was infused into the duodenum and capsule disintegration occurred early, there was significant dilution and spreading of the excipients throughout a large section of the upper small intestine (USI). However, when the radiopaque dye was infused into the ileum and capsule disintegration occurred in the lower small intestine (LSI), the excipients moved along as a bolus (i.e., plug). The pH monitoring results were consistent with the fluoroscopy results. The pH dropped only momentarily and rose quickly in the USI consistent with well-stirred mixing kinetics. In the LSI, dilution and spreading were minimal and the drop in pH was greater and persisted for a longer period of time. Plasma levels of sCT were maximal when disintegration occurred in the LSI. CONCLUSIONS: Since significantly less dilution and spreading occurred in the LSI, the exposure of the intestine to pharmaceutical excipients and sCT was more concentrated resulting in a higher fraction of sCT absorbed. The results of this study demonstrate that intestinal mixing kinetics have a dramatic impact on the ability of pharmaceutical excipients to modulate the oral bioavailability of peptide drugs like sCT. PMID- 10801217 TI - Route-dependent metabolism of morphine in the vascularly perfused rat small intestine preparation. AB - PURPOSE: 1. To compare the disposition of tracer morphine ([3H]M) following systemic and intraduodenal administration in the recirculating, rat small intestine preparation in absence or presence of verapamil (V), an inhibitor of P glycoprotein. 2. To develop a physiological model to explain the observations. METHODS: A bolus dose of [3H]M was added to the reservoir or injected into the duodenum of the rat small intestine preparation. V (200 microM in reservoir) was either absent (control studies) or present. Intestinal microsomal, incubation studies were performed to evaluate the effect of V on morphine glucuronidation. RESULTS: After systemic administration, [3H]M was not metabolized but was exsorbed into lumen. By contrast, both [3H]M and the 3beta-glucuronide metabolite, [3H]M3G, appeared in reservoir and lumen after intraduodenal administration. A physiologically-based model that encompassed absorption, metabolism and secretion was able to describe the route-dependent glucuronidation of M. The presence of V resulted in diminished levels of M3G in perfusate and lumen and mirrored the observation of decreased glucuronidation in microsomal incubations. Verapamil appeared to be an inhibitor of glucuronidation and not secretion of M. CONCLUSIONS: M was secreted and absorbed by the rat small intestine. Route-dependent glucuronidation of M was explained by physiological modeling when M was poorly partitioned in intestinal tissue, with a low influx clearance from blood and a even poorer efflux clearance from tissue. The poor efflux rendered a much greater metabolism of M that was initially absorbed from the lumen. V increased the extent of M absorption through inhibition of M glucuronidation. PMID- 10801218 TI - Persistent inhibition of CYP3A4 by ketoconazole in modified Caco-2 cells. AB - PURPOSE: The intestinal metabolism of some CYP3A substrates can be altered profoundly by co-administration of the potent inhibitor, ketoconazole. The present research was conducted to test the hypothesis that, unlike the inhibition kinetics observed with isolated microsomes, inhibition of CYP3A4 by ketoconazole in an intestinal cell monolayer is time-dependent and slowly reversible. METHODS: Confluent, 1alpha,25-dihydroxy Vitamin D3-treated Caco-2 cells were exposed to 1 microM ketoconazole for two hours (Phase I) and then washed three times with culture medium containing no inhibitor. This was followed by a second incubation period (Phase II) that varied in the composition of the apical and basolateral culture medium: Condition 1. apical/basolateral differentiation medium (DM); Condition 2, apical/ basolateral DM + basolateral 2g/dL Human Serum Albumin (HSA); Condition 3, apical/basolateral DM + apical/basolateral 2 g/dL HSA. After various lengths of time for the second phase (0 to 4 hours), both apical and basolateral medium were exchanged with fresh DM. Midazolam (6 microM) was included in the apical medium for determination of CYP3A4 activity (Phase III). RESULTS: Two-way ANOVA of the data revealed persistent inhibition of CYP3A4 under Conditions 1 and 2 (p < 0.001). In contrast, cells treated under Condition 3 exhibited rapid reversal of CYP3A4 inhibition. The level of CYP3A4 activity observed was inversely correlated with the amount of ketoconazole remaining in the cell monolayer at the end of Phase II. CONCLUSIONS: These studies provide mechanistic evidence that ketoconazole can be sequestered into the intestinal mucosa after oral administration, producing a persistent inhibition of first-pass CYP3A4 activity. PMID- 10801219 TI - In vivo gene delivery to the liver using novel galactosylated cationic liposomes. AB - PURPOSE: The purpose of this study is to elucidate the in vivo gene transfer for galactosylated liposomes containing cholesten-5-yloxy-N-(4-((1-imino-2-beta-D thiogalactosyle thyl)amino)butyl)formamide(Gal-C4-Chol) in relation to lipid composition and charge ratio. METHODS: Galactosylated cationic liposomes containing N-[1-(2,3-dioleyloxy)propyl]-n,n,n-trimethylammonium chloride(DOTMA), Gal-C4-Chol and cholesterol(Chol), and similar liposomes were prepared. Plasmid DNA complexed with a galactosylated liposome preparation was injected intraportally into mice. The mice were sacrificed after 6 hours. The tissues were subjected to luciferase assay. RESULTS: A markedly higher gene expression in the liver following injection of plasmid DNA that has been complexed with DOTMA/ Chol/Gal-C4-Chol(1:0.5:0.5) and DOTMA/Gal-C4-Chol(1:1) liposomes was observed. The effect was one order of magnitude higher than naked DNA and DOTMA/Chol(1:1) liposomes. Pre-exposing with galactosylated bovine serum albumin significantly reduced the hepatic gene expression. By comparison, the gene expression for galactosylated cationic liposomes containing 3beta[N-(N',N' dimethylaminoethane)carbamoyl]cholesterol, Gal-C4-Chol and dioleoylphosphatidylethanolamine was 10 times lower. As far as the charge ratio of DOTMA/ Chol/Gal-CA-Chol(1:0.5:0.5) liposomes to plasmid DNA(1.6-7.0) was concerned, complexes with charge ratios of 2.3-3.1 produced maximal gene expression in the liver. Whereas, higher ratios resulted in enhanced expression in the lung. CONCLUSIONS: By optimizing lipid composition and charge ratio, galactosylated liposome/DNA complexes allow superior in vivo gene transfection in the liver via asialoglycoprotein receptor-mediated endocytosis. PMID- 10801220 TI - A cationic lipid emulsion/DNA complex as a physically stable and serum-resistant gene delivery system. AB - PURPOSE: To develop a non-viral gene delivery system in the form of an oil-in water (o/w) lipid emulsion. METHOD: Cationic lipid emulsions were formulated with soybean oil, 1,2-dioleoyl-sn-glycero-3-trimethylammonium-propane (DOTAP) as a cationic emulsifier and other co-emulsifiers. The physical characteristics of the lipid emulsion and the emulsion/DNA complex were determined. The in vitro transfection efficiency of the emulsion/DNA complex was determined in the presence of up to 90% serum. RESULTS: The average droplet size and zeta potential of emulsions were ca. 180 nm and ca. +50 mV, respectively. Among the emulsions, a stable formulation was selected to form a complex with a plasmid DNA encoding chloramphenicol acetyltransferase. By increasing the ratio of emulsion to DNA. zeta-potential of the emulsion/DNA complex increased monotonously from negative to positive without any changes in the complex size. The complex was stable against DNase I digestion and an anionic poly-L-aspartic acid (PLAA). The complex delivered DNA into the cells successfully, and the transfection efficiency was not affected by complex formation time from 20 min to 2 h. More importantly, the cationic lipid emulsion facilitated the transfer of DNA in the presence of up to 90% serum. CONCLUSIONS: The cationic lipid emulsion/DNA complex has physical stability and serum resistant properties for gene transfer. PMID- 10801221 TI - Mechanism-based modeling of functional adaptation upon chronic treatment with midazolam. AB - PURPOSE: A mechanism-based model is applied to analyse adaptive changes in the pharmacodynamics of benzodiazepines upon chronic treatment in rats. METHODS: The pharmacodynamics of midazolam was studied in rats which received a constant rate infusion of the drug for 14 days, resulting in a steady-state concentration of 102 +/- 8 ng x ml(-1). Vehicle treated rats were used as controls. Concentration EEG effect data were analysed on basis of the operational model of agonism. The results were compared to data obtained in vitro in a brain synaptoneurosomal preparation. RESULTS: The relationship between midazolam concentration and EEG effect was non-linear. In midazolam pre-treated rats the maximum EEG effect was reduced by 51 +/- 23 microV from the original value of 109 +/-15 microV in vehicle treated group. Analysis of this change on basis of the operational model of agonism showed that it can be explained by a change in the parameter tissue maximum (Em) rather than efficacy (tau). In the in vitro studies no changes in density, affinity or functionality of the benzodiazepine receptor were observed. CONCLUSIONS: It is concluded that the observed changes in the concentration-EEG effect relationship of midazolam upon chronic treatment are unrelated to changes in benzodiazepine receptor function. PMID- 10801222 TI - Population pharmacokinetic/pharmacodynamic modeling of cetrorelix, a novel LH-RH antagonist, and testosterone in rats and dogs. AB - PURPOSE: Population models for the pharmacokinetic-pharmacodynamic relationship for cetrorelix (CET), a luteinising hormone-releasing hormone (LH-RH) antagonist, and the pharmacodynamic response on testosterone production were investigated in rats and dogs. METHODS: The plasma concentrations of CET and testosterone were determined after intravenous and subcutaneous injections. The population PK/PD models were developed using P-PHARM software. RESULTS: Absolute bioavailability of cetrorelix was 100% in rats and 97% in dogs. In rats, the pharmacokinetics was explained by a two-compartment model with saturable absorption, while a three compartment model was used in dogs. Testosterone suppression in both species was described by a sigmoid E(max) model with maximum effect (E(max)) considered as total hormonal suppression. The duration of testosterone suppression in rats was longer at higher doses. The population elimination half-lives after iv-dose were 3.0 h in rats and 9.3 h in dogs. Population mean estimates of IC50 were 1.39 and 1.24 ng/ml in rats and dogs, respectively. CONCLUSIONS: A population pharmacokinetic model was developed to explain the dissolution rate limited absorption from the injection site. The suppression of testosterone could be described by an indirect inhibitory sigmoid E(max) model. In both species 1-2 ng/ml CET in plasma was necessary to suppress testosterone production. PMID- 10801223 TI - Quantitative prediction of in vivo drug-drug interactions from in vitro data based on physiological pharmacokinetics: use of maximum unbound concentration of inhibitor at the inlet to the liver. AB - PURPOSE: To assess the degree to which the maximum unbound concentration of inhibitor at the inlet to the liver (I(inlet,u,max), used in the prediction of drug-drug interactions, overestimates the unbound concentration in the liver. METHODS: The estimated value of I(inlet,u,max) was compared with the unbound concentrations in systemic blood, liver, and inlet to the liver, obtained in a simulation study based on a physiological flow model. As an example, a tolbutamide/sulfaphenazole interaction was predicted taking the plasma concentration profile of the inhibitor into consideration. RESULTS: The value of I(inlet,u,max) differed from the concentration in each compartment, depending on the intrinsic metabolic clearance in the liver, first-order absorption rate constant, non-hepatic clearance and liver-to-blood concentration ratio (Kp) of the inhibitor. The AUC of tolbutamide was predicted to increase 4-fold when co administered with sulfaphenazole, which agreed well with in vivo observations and was comparable with the predictions based on a fixed value of I(inlet,u,max). The blood concentration of tolbutamide was predicted to increase when it was co administered with as little as 1/100 of the clinical dose of sulfaphenazole. CONCLUSIONS: Although I(inlet,u,max) overestimated the unbound concentration in the liver, the tolbutamide/sulfaphenazole interaction could be successfully predicted by using a fixed value of I(inlet,u,max) indicating that the unbound concentration of sulfaphenazole in the liver after its clinical dose is by far larger than the concentration to inhibit CYP2C9-mediated metabolism and that care should be taken when it is co-administered with drugs that are substrates of CYP2C9. PMID- 10801224 TI - Liposomes as formulation excipients for protein pharmaceuticals: a model protein study. AB - PURPOSE: The advent of recombinant DNA technology has made possible the pharmaceutical use of a wide range of proteins and peptides. However, the complex structure of proteins renders them susceptible to physical instabilities such as denaturation, aggregation and precipitation. We tested the hypothesis that partial unfolding and exposure of hydrophobic domains leads to physical instability, and investigated approaches to stabilize protein formulations. METHODS: KP6 beta, an 81 amino acid killer toxin from Ustilago maydis, was used as a model protean. Circular dichroism and fluorescence spectroscopy were used to study the temperature dependent folding/ unfolding characteristics of KP6 beta. ANS (1,8 anilinonaphthalene sulfonate), a fluorescent probe that partitions into hydrophobic domains, was used to detect exposure of hydrophobic domains. RESULTS: As the temperature was elevated, near-UV CD indicated progressive loss of KP6 beta tertiary structure, while far-UV CD indicated retention of secondary structure. Increasing exposure of hydrophobic domains was observed, as indicated by the penetration of ANS. At elevated temperatures (60 degrees C), KP6 beta3 conserved most secondary structural features. However, tertiary structure was disordered, suggesting the existence of a partially folded, structured intermediate state. Liposomes bound to partially unfolded structures and prevented the formation of aggregates. CONCLUSIONS: Partial unfolding resulted in increased exposure of hydrophobic domains and aggregation of KP6 beta, but with preservation of secondary structure. Liposomes interacted with the structured intermediate state, stabilizing the protein against aggregation. These results suggest a general formulation strategy for proteins, in which partially unfolded structures are stabilized by formulation excipients that act as molecular chaperones to avoid physical instability. PMID- 10801225 TI - Stabilization of proteins encapsulated in cylindrical poly(lactide-co-glycolide) implants: mechanism of stabilization by basic additives. AB - PURPOSE: A previous study from our group has shown that in the acidic microclimate of poly(lactide-co-glycolide) (PLGA) implants, encapsulated BSA forms insoluble noncovalent aggregates and is hydrolyzed during in vitro release. Incorporation of Mg(OH)2 strongly inhibits these mechanisms of instability and facilitates continuous protein release. The purpose of this study was to determine the protein stabilization mechanism in the presence of basic additives. METHODS: BSA, as a model protein, was encapsulated in PLGA millicylinders by a solvent extrusion method. The release of BSA from the PLGA millicylinders with and without basic additives (Mg(OH)2, Ca(OH)2, ZnCO3 and Ca3(PO4)2) in a physiological buffer was carried out at 37 degrees C and quantified by a modified Bradford assay. The insoluble aggregates extracted from the polymer with acetone were reconstituted in a denaturing (6 M urea) or denaturing/reducing solvent (6 M urea/10 mM DTT) to determine the type of aggregation. RESULTS: Aggregation of encapsulated BSA was inhibited with increasing amount of base co-encapsulated in the polymer, irrespective of the type of base used. The pH drop in the release medium and extent of acid-catalyzed PLGA degradation were both inhibited in the presence of base. The resultant effect was also reflected in an increase in water uptake and porosity of the devices. The inhibition and mechanism of BSA aggregation was correlated with the basicity of the additive. For Ca(OH)2, at 3% loading, covalent BSA aggregation due to thioldisulfide interchange was observed (indicative of ionization of albumin's free thiol at high pH), whereas at 3% ZnCO3 or Ca3(PO4)2, a higher percentage of non-covalent aggregates was observed compared to Mg(OH)2. Decreasing the loading of BSA at constant Mg(OH)2 content caused an increase in BSA aggregation. CONCLUSIONS: The mechanism by which Mg(OH)2 stabilizes encapsulated BSA in PLGA implants is through neutralizing the acidic microclimate pH in the polymer. The successful neutralization afforded by the basic additives requires a percolating network of pores connecting both base and protein. The microclimate pH inside PLGA implants can be controlled by selecting the type of basic salt, which suggests a potential approach to optimize the stability of encapsulated pharmaceuticals in PLGA including therapeutic proteins. PMID- 10801226 TI - Predicting the free energies of complexation between cyclodextrins and guest molecules: linear versus nonlinear models. AB - PURPOSE: In the present paper, linear and nonlinear models for complexation of alpha- beta- and gamma-cyclodextrin with guest molecules are developed, with the aim of free energy prediction and interpretation of the association process. METHODS: Linear and nonlinear regression is used to correlate experimental free energies of complexation with calculated molecular descriptors. Molecular modeling supports the interpretation of the results. RESULTS: Highly predictive models are obtained, although the structural variability of the compounds used for their deduction is large, reaching from synthetic heterocycles to steroids and prostaglandins. CONCLUSIONS: The scaled regression coefficients give insight to the complexation mechanisms, which appear to be different for the three types of cyclodextrins. PMID- 10801227 TI - Acid-catalyzed inversion of sucrose in the amorphous state at very low levels of residual water. AB - PURPOSE: Factors affecting the solid-state acid-catalyzed inversion of amorphous sucrose to glucose and fructose in the presence of colyophilized citric acid, with less than 0.1% w/w residual water, have been studied. METHODS: Samples of citric acid and sucrose were lyophilized at a weight ratio of 1:10 citric acid:sucrose from solutions with initial pH values of 1.87, 2.03, and 2.43, as well as at a weight ratio of 1:5, at an initial pH of 1.87. Glass transition temperatures, Tg, were measured by DSC and the presence of any possible residual water was monitored by Karl Fischer Titrimetry. The inversion of sucrose was measured by polarimetric analysis after reconstitution of solid samples stored at 50 degrees C under P2O5. RESULTS: Samples of 1:10 citric acid:sucrose at an initial pH of 1.87, 2.03, and 2.43 exhibited the same Tg. The initial rate of reactivity was affected at a 1:10 ratio by the solution pH before lyophilization in the order: 1.87 > 2.03 > 2.43 and by citric acid concentration at pH 1.87 in the order 1:5 > 1:10. CONCLUSIONS: Sucrose, colyophilized with an acid such as citric acid, undergoes significant acid-catalyzed inversion at 50 degrees C despite the very low levels of residual water, i.e., <0.1% w/w. At the same ratio of citric acid to sucrose (1:10), and hence the same Tg, the rate of reaction correlates with the initial solution pH indicating that the degree of ionization of citric acid in solution is most likely retained in the solid state. That protonation of sucrose by citric acid is important is shown by the direct relationship between maximum extent of reaction and citric acid composition. It is concluded that colyophilization of acidic substances with sucrose, even in the absence of residual water, can produce reducing sugars capable of further reaction with other formulation ingredients susceptible to reaction with reducing sugars. PMID- 10801228 TI - Recognition of nonpeptide prenyl pyrophosphate antigens by human gammadelta T cells. PMID- 10801229 TI - Gammadelta T cells and Mycobacterium tuberculosis. AB - Since the first descriptions of mycobacterial reactivity for gammadelta T cells in 1989, studies of gammadelta T-cell responses to M. tuberculosis in humans and animal models have increased our understanding of the complex role(s) of this T cell subset not only in the immune response to M. tuberculosis, but also to microbial pathogens in general. Although CD4+ T cells remain the dominant and critical T-cell subset in protection against M. tuberculosis, gammadelta T cells appear to have an important complementary role, which may be primarily expressed in and around maturing granulomas. This is a difficult area to study in humans. Gammadelta T cells are potent sources of IFN-gamma and competent cytotoxic effector cells, but differ from CD4+ T cells in the antigens they recognize and the manner in which M. tuberculosis-infected macrophages process and present antigens to these two subsets. One of the most fascinating features of Vgamma9/Vgamma2+ gammadelta T cells is their responsiveness to non-peptidic molecules. Solving the mechanism(s) of antigen recognition and presentation of these molecules to gammadelta T cells should help determine whether gammadelta T cells are responding to universal 'supernatigen'-like motifs expressed by a broad range of microbes or in fact discriminate among a diversity of peptidic and nonpeptidic microbial antigens. Enhanced understanding of the function of and antigen recognition by Vgamma9+/Vgamma2+ T cells is not only important for immunity to M. tuberculosis but also for T-cell responses to microbial pathogens in general. PMID- 10801230 TI - Activation of gammadelta T cells by mycobacterial antigens in rheumatoid arthritis. PMID- 10801231 TI - TCRgammadelta cells and viruses. AB - T-cell receptor gammadelta cells (TCRgammadelta) are often found in increased numbers during the course of several viral infections in humans. Although these findings suggest an important role for this unique subset, their precise function has not been ascertained. Recent studies in murine models of both RNA and DNA virus infections have begun to shed new light on the potential function for TCRgammadelta cells in antiviral immunity. It is clear that TCRgammadelta cells participate in the immune response to human immunodeficiency virus (HIV), influenza, Sendai, coxsackie, vaccinia, vesicular stomatitis virus (VSV), and herpes simplex virus-1 (HSV-1) viral infections since they become activated and home to the sites of viral replication. In this review we will summarize current efforts to dissect the role of TCRgammadelta cells in these disease settings, emphasizing the effector functions utilized, the TCR repertoire, and the antigens recognized. Particular focus will be placed on HSV-1 infections where we have begun to address these issues and have shown that TCRgammadelta cells are sufficient for protection from lethal infection and are able to recognize the herpes virus antigen glycoprotein I. PMID- 10801232 TI - Human gammadelta T cells and viruses. PMID- 10801233 TI - NKR-mediated control of gammadelta T-cell immunity to viruses. PMID- 10801234 TI - Role of gammadelta T cells in tumor immunity and their control by NK receptors. PMID- 10801235 TI - The role of intraepithelial gammadelta T cells: a gut-feeling. PMID- 10801236 TI - Immunoregulatory functions of mucosal gammadelta T cells. PMID- 10801237 TI - Activation and control of self-reactive gammadelta T cells. AB - Mammalian and avian CD3+ T cells can be separated into two lymphocyte subsets bearing heterodimeric T-cell receptors (TCR) composed of either alphabeta or gammadelta chains. Although it is now widely accepted that gammadelta and alphabeta T cells fulfill mandatory and nonredundant roles, the generality of this assumption and the exact functions played by gammadelta T cells remain uncertain. While an early protective role of gammadelta T cells has long been suspected, recent observations drawn in particular from transgenic models suggest their implication in the homeostatic control of immune and nonimmune processes. This hypothesis is also supported by the existence of several self-reactive gammadelta T-cell subsets in rodents and humans, whose specificity and effector properties will be detailed and discussed here. The present review will also describe several mechanisms that could allow efficient control of these self reactive subsets while permitting expression of their regulatory and/or protective properties. PMID- 10801238 TI - Effector functions and control of human gammadelta T-cell activation. PMID- 10801239 TI - Control of gammadelta T cells by NK receptors. PMID- 10801240 TI - Erratic patient compliance with prescribed drug regimens: target for drug delivery systems. PMID- 10801241 TI - Alprazolam-ritonavir interaction: implications for product labeling. AB - BACKGROUND: Pharmacokinetic interactions involving antiretroviral therapies may critically influence the efficacy and toxicity of these drugs, as well as pharmacologic treatments of coincident or complicating diseases. The viral protease inhibitor ritonavir is of particular concern since it both inhibits and induces the activity of cytochrome P450 3A (CYP3A) isoforms. METHODS: The inhibitory effect of ritonavir on the metabolism of alprazolam, a CYP3A-mediated reaction in humans, was tested in vitro using human liver microsomes. In a double blind clinical study, volunteer subjects received 1.0 mg of alprazolam concurrent with low-dose ritonavir (four doses of 200 mg) or with placebo. RESULTS: Ritonavir was a potent in vitro inhibitor of alprazolam hydroxylation. The 50% inhibitory concentration was 0.11 micromol/L (0.08 microg/mL); this is below the usual therapeutic plasma concentration range (generally exceeding 2 microg/mL). In the clinical study, ritonavir reduced alprazolam clearance to 41% of control values (P < .001), prolonged elimination half-life (mean values, 30 versus 13 hours; P < .005), and magnified benzodiazepine agonist effects such as sedation and performance impairment. CONCLUSION: Consistent with in vitro results, administration of low doses of ritonavir for a short duration of time resulted in large impairment of alprazolam clearance and enhancement of clinical effects. Removal from product labeling of a warning against coadministration of ritonavir and alprazolam was based on a previous study only of extended exposure to ritonavir, in which CYP3A induction offset inhibition. Kinetic interactions involving antiretroviral therapies may be complex and time dependent. Product labeling should reflect this complexity. PMID- 10801242 TI - Indocyanine green kinetics characterize blood volume and flow distribution and their alteration by propranolol. AB - BACKGROUND AND OBJECTIVES: Although indocyanine green can be used to estimate cardiac output and blood volume independently, a recirculatory multicompartmental indocyanine green model enables description of these and additional intravascular events. Our model was used to describe the effect of propranolol on blood volume and flow distribution in humans. METHODS: Indocyanine green disposition was determined twice in four healthy adult men, once during a propranolol infusion that decreased cardiac output. After injection of indocyanine green, arterial blood was collected frequently for 2 minutes and less frequently thereafter. Plasma indocyanine green concentrations were measured by HPLC. The recirculatory pharmacokinetic model incorporates data from both the initial transient oscillations and the later post-mixing portions of the blood indocyanine green concentration versus time curves to characterize not only blood volume and cardiac output but also their distribution among a central blood volume and fast and slow peripheral volumes in lumped parallel circuits. Flow through the central circulation (cardiac output) is described by two parallel Erlang distribution functions generated by two linear chains of compartments in parallel. RESULTS: Propranolol reduced cardiac output from 10.6 to 4.1 L/min. Most of the decrease in cardiac output was at the expense of blood flow to the fast peripheral circuit, which represented nonsplanchnic circulation. Propranolol also reduced the blood volume of the fast peripheral circuit by more than half. CONCLUSION: Our indocyanine green model is able to derive estimates of blood volume and cardiac output, as well as their systemic distribution during different physiologic conditions. PMID- 10801243 TI - The steady-state disposition of indinavir is not altered by the concomitant administration of clarithromycin. AB - STUDY OBJECTIVES: To evaluate the safety and potential pharmacokinetic interaction between indinavir and clarithromycin. STUDY METHODS: In a randomized, three-period, crossover fashion, 12 healthy adults received the following for 1 week: 800 mg oral indinavir sulfate every 8 hours with placebo, 500 mg oral clarithromycin every 12 hours with placebo, and indinavir sulfate with clarithromycin. Plasma indinavir, clarithromycin, and 14-hydroxyclarithromycin concentrations were determined after the last dose in each treatment period. RESULTS: Administration of indinavir sulfate with clarithromycin caused a statistically significant increase in four pharmacokinetic parameters: a 58% increase in plasma indinavir concentrations at 8 hours (P = .029), a 47% increase in values for clarithromycin area under the plasma concentration versus time curve from time zero to the last measured concentration [AUC(0-12h); P = .0002], and 49% and 48% decreases in 14-hydroxyclarithromycin AUC(0-12h) and maximum plasma concentration (Cmax) values, respectively (P = .0001 and P = .0001). These effects are not considered to be clinically significant in view of the insignificant effects on the values for indinavir area under the plasma concentration versus time curve from time zero to the last measured concentration [AUC(0-8h)] and Cmax, as well as the safety profile of clarithromycin. CONCLUSIONS: The combination of indinavir sulfate and clarithromycin is generally well tolerated and can be coadministered without dose adjustment. PMID- 10801245 TI - Pharmacokinetics of cefpirome during continuous venovenous hemofiltration: rationale for an 8-hour dosing interval. AB - OBJECTIVE: Cefpirome is a new semisynthetic cephalosporin, primarily eliminated by the kidneys, that requires dosage adjustment in patients with kidney failure. The optimal dosing regimen of cefpirome in patients with continuous veno-venous hemofiltration (CVVH) is unknown. METHODS: Pharmacokinetic properties of cefpirome were investigated in eight anuric patients with acute kidney failure treated by CVVH. All patients received a dosage of 2 g cefpirome every 8 hours after starting the hemofiltration with high-flux polysulfone membranes. Concentrations of cefpirome in plasma and ultrafiltrate were measured by HPLC. RESULTS: Total clearance and hemofiltration clearance of cefpirome were 589.1 +/- 164.5 mL/min and 43.3 +/- 7.8 mL/min, respectively. Serum elimination half-life was 2.36 +/- 0.59 hours. The highest plasma drug concentration was 14.8 +/- 3.2 microg/mL, and it declined to trough levels of 3.1 +/- 0.8 microg/mL at the end of the dosing interval. CONCLUSION: On the basis of previously published pharmacodynamic characteristics of cefpirome and the pharmacokinetic parameters obtained in this study, we calculated a required total daily dose of 2 g every 8 hours to achieve sufficient plasma antibiotic levels to cover the majority of target pathogens. However, this dosage may be insufficient during CVVH for intermediate resistant strains of Pseudomonas aeruginosa. PMID- 10801244 TI - Vancomycin population pharmacokinetics in neonates. AB - BACKGROUND: Recently the value of vancomycin therapeutic drug monitoring, as well as the required therapeutic range, has been subject of debate, resulting in new recommendations. This study was performed to incorporate these new insights in an up-to-date dosing scheme for neonates of various gestational ages. METHODS: In this retrospective study with prospective validation, 108 newborns with suspected central line-related septicemia during the first month of life received 30 mg/kg/day vancomycin divided into two doses regardless of gestational or postconceptional age. Trough and peak vancomycin serum concentrations were determined before and after the third dose. Vancomycin data were analyzed according to a one-compartment open model with use of NONMEM population pharmacokinetic software. Model parameters were evaluated and then used to simulate vancomycin dosing for different dose and dose interval combinations. Targets were a trough concentration between 5 and 15 mg/L and a peak below 40 mg/L. In the prospective study, the optimal scheme was tested in 22 patients. RESULTS: Of the 108 patients, 34.3% of measured trough concentrations and 17.6% of peak concentrations were outside the desired therapeutic range. The model that best fitted the data included clearance and volume per kilogram and was independent of gestational age. Simulation of various dosing schemes showed that a dosing schedule of 30 mg/kg/day, irrespective of gestational age, in three doses was optimal, and this scheme was prospectively tested. Mean trough concentrations before the second dose were 8.2 +/- 2.2 mg/L versus a predicted trough of 8.9 +/- 2.5 mg/L. No peak levels higher than 40 mg/L were found. CONCLUSIONS: The use of the proposed schedule leads to adequate vancomycin trough serum concentrations, and there is no need for routine monitoring of peak serum concentrations. PMID- 10801246 TI - Effect of an oral contraceptive on the plasma levels of budesonide and prednisolone and the influence on plasma cortisol. AB - OBJECTIVE: To investigate whether the use of an oral contraceptive would influence plasma levels of budesonide (Entocort capsules) or prednisolone (plain tablets) during repeated oral administration of these glucocorticosteroids. Plasma concentrations of cortisol and ethinyl estradiol (INN, ethinylestradiol) were also compared. METHODS: Forty healthy women took part in this single-blind, randomized placebo-controlled study with two parallel groups, where a three-way crossover design was applied within groups. One group was taking an oral contraceptive (150 microg desogestrel and 30 microg ethinyl estradiol); the other group (control) was not. On seven consecutive mornings, oral doses of 4.5 mg budesonide, 20 mg prednisolone, or placebo were administered. There was a washout period of at least one menstrual cycle between administration periods. RESULTS: In the oral contraceptive users, the average plasma concentration of prednisolone was 131% higher than in the control group (P < .001), whereas the average plasma concentration of budesonide was only 22% higher (not significant). Mean plasma cortisol levels were suppressed by 90% and 82% with prednisolone and by 22% and 28% with budesonide in oral contraceptive users and the control subjects, respectively. The group difference was significant with prednisolone (P < .001) but not with budesonide. Ethinyl estradiol levels in plasma were not affected by administration of either glucocorticosteroid. CONCLUSION: No difference was found in plasma levels of budesonide or in cortisol suppression after administration of budesonide capsules in women taking the oral contraceptive and those who were not. The oral contraceptive users had much higher plasma levels of prednisolone and greater cortisol suppression. This result suggests that oral budesonide can be used with maintained safety in women using oral contraceptives. PMID- 10801247 TI - Lack of correlation between in vitro and in vivo studies on the effects of tangeretin and tangerine juice on midazolam hydroxylation. AB - BACKGROUND: Tangeretin is a flavonoid that stimulates the catalytic activity of cytochrome P450 3A4 (CYP3A4) and is found in high levels in tangerine juice. METHODS: The effect of tangeretin on hydroxylation of midazolam, a CYP3A4 probe, was examined in vitro with human liver microsomes and recombinant CYP3A4. In addition, the effect of tangerine juice on the pharmacokinetics and pharmacodynamics of orally administered midazolam (15 mg) and its active 1' hydroxymetabolite was studied in a randomized crossover study in eight healthy volunteers. RESULTS: In microsomes from three human livers, tangeretin (1 to 100 micromol/L) increased 1'-hydroxymidazolam formation (12.5 micromol/L midazolam) by up to 212%. In complementary deoxyribonucleic acid-expressed CYP3A4, a 52% stimulation of midazolam 1'-hydroxylation was reached at 50 micromol/L tangeretin with no effect on midazolam 4-hydroxylation. In the pharmacokinetic pharmacodynamic study, 200 mL tangerine juice reduced the area under the concentration versus time curve to 1.5 hours [AUC(O-1.5h)] of midazolam and 1' hydroxymidazolam by 39% and 46%, respectively, and prolonged the time to reach peak concentration (P < .05) without affecting the total AUC values, elimination half-life values, or AUC ratios (1'-hydroxymidazolam/midazolam). These findings are consistent with a small delay in the absorption of midazolam and lack of effect on midazolam 1'-hydroxylation. Accordingly, tangerine juice slightly postponed the maximum pharmacodynamic effects of midazolam (P < .05). CONCLUSION: Tangeretin is a potent regioselective stimulator of midazolam 1'-hydroxylation by human liver microsomes and complementary deoxyribonucleic acid-expressed CYP3A4. However, tangerine juice is unlikely to have any appreciable effect on CYP3A4 in humans. Further studies are required to assess whether in vitro stimulators of CYP3A4 can influence drug metabolism in vivo. PMID- 10801248 TI - Nicotine impairs endothelium-dependent dilatation in human veins in vivo. AB - BACKGROUND: Cigarette smoking is associated with impaired endothelium-dependent dilatation in human veins and arteries. An in vivo study in animals suggests that nicotine may contribute to this abnormality. We tested the hypothesis that local administration of nicotine at a dose reproducing the plasma concentration observed during smoking would impair endothelium-dependent vasodilatation in human veins in vivo. METHODS: We studied 11 healthy nonsmokers with the dorsal hand vein compliance technique. After 70% to 80% preconstriction with phenylephrine, endothelium-dependent venous relaxation was assessed by infusion of bradykinin (1 to 278 ng/min), a potent vasodilator acting primarily in this model through endothelial release of nitric oxide and prostanoids. Sodium nitroprusside (0.0001 to 3166 ng/min) was used to test endothelium-independent relaxation. Dose-response curves were constructed before and during nicotine coinfusion at a rate of 40 ng/min, reproducing a plasma concentration of 15 ng/mL. RESULTS: After a 10-minute preinfusion, nicotine administration was associated with a loss in sensitivity to bradykinin (P < .001). After 30 and 60 minutes of preinfusion with nicotine, the venorelaxant effect of bradykinin was further reduced (P < .001). A similar inhibition of the response to bradykinin by nicotine persisted in the presence of indomethacin (INN, indomethacin). Coinfusion of nicotine did not attenuate sodium nitroprusside-induced venodiiation. CONCLUSION: The results show that acute local exposure to nicotine in vivo is associated with an impaired response to endothelium-derived nitric oxide in human veins. This finding may provide further insight into the pathophysiology of smoking-induced endothelial dysfunction. PMID- 10801249 TI - Effects of enteric-coated methylnaltrexone in preventing opioid-induced delay in oral-cecal transit time. AB - BACKGROUND: Methylnaltrexone is the first peripheral opioid receptor antagonist. It has the potential to prevent or reverse the peripherally mediated gastrointestinal effects of opioids. In previous human volunteer trials, we demonstrated that oral uncoated methylnaltrexone prevented morphine-induced delay in gastrointestinal transit time. METHODS: This trial consisted of two studies: a pilot study and a controlled study. The lactulose hydrogen breath test was used to measure the oral-cecal transit time. RESULTS: In the pilot study with three subjects, an oral dose of 6.4 mg/kg enteric-coated methylnaltrexone effectively reversed the effects of morphine, producing transit times shorter than baseline levels. Subsequently, in the controlled study with another nine subjects, the transit time increased after intravenous morphine administration in all nine subjects, and the lower dose (3.2 mg/kg) of enteric-coated methylnaltrexone completely prevented the morphine-induced change in oral-cecal transit time in all nine subjects. Morphine significantly increased oral-cecal transit time from baseline level of 96.7 +/- 54.1 minutes (mean +/- SD) to 155.0 +/- 53.6 minutes (P = .014). After enteric-coated methylnaltrexone and morphine, the transit time returned to the baseline level (93.3 +/- 56.0 minutes; P = .55 compared with placebo). Plasma concentrations after 6.4 mg/kg and 3.2 mg/kg enteric-coated methylnaltrexone were substantially lower compared with those after 6.4 mg/kg of the uncoated formulation. CONCLUSION: Our results suggest that there is a prevailing direct and local luminal effect of enteric-coated methylnaltrexone and that the enteric-coated formulation exerts its gut pharmacologic actions more efficiently than the uncoated formulation. PMID- 10801250 TI - Antagonistic activity of tamsulosin against human vascular alpha1-adrenergic receptors. AB - OBJECTIVE: To elucidate the vascular effect of tamsulosin hydrochloride (INN, tamsulosin), a selective alpha1A-adrenergic receptor antagonist, in humans, we examined the alpha1-adrenergic receptor antagonistic activity against blood vessels after oral intake of recommended and higher doses of the drug and evaluated the relation between its plasma concentrations and the effect. METHODS: Nine healthy men ranging in age from 21 to 38 years received tamsulosin (0.2 mg or 0.6 mg) or lactate capsule as a control after breakfast in a randomized crossover fashion. Seven hours after administration, their fingertip vasoconstrictor response to contralateral hand cooling and vasoconstrictor response of the dorsal hand vein to phenylephrine were examined, and blood samples for the measurement of plasma drug concentration were obtained. RESULTS: The fingertip vasoconstrictor response was significantly reduced and the infusion rate of phenylephrine producing a half-maximal constriction was significantly increased by 0.6 mg tamsulosin but not by 0.2 mg tamsulosin. There were significant positive correlations between plasma drug concentrations and the changes of these parameters. CONCLUSION: These results suggest that although the alpha1-adrenergic receptor-blocking effect of tamsulosin on blood vessels is relatively small, it is clearly correlated with plasma drug concentration and a higher dose of the drug could cause systemic adverse effects. PMID- 10801251 TI - Greater quinidine-induced QTc interval prolongation in women. AB - BACKGROUND: Prolongation of the electrocardiographic QT interval by drugs is associated with the occurrence of a potentially lethal form of polymorphic ventricular tachycardia termed torsades de pointes. Women are at greater risk than men for development of this adverse event when taking drugs that prolong the QT interval. To determine whether this may be the result of gender-specific differences in the effect of quinidine on cardiac repolarization, we compared the degree of quinidine-induced QT interval lengthening in healthy young men and women. METHODS: Twelve women and 12 men received a single intravenous dose of quinidine (4 mg/kg) or placebo in a single-blind, randomized crossover trial. Total plasma and protein-free concentrations of quinidine and 3-hydroxyquinidine were measured in serum. QT intervals were determined and corrected for differences in heart rate with use of the method of Bazett (QTc = QT/RR1/2). RESULTS: As expected, the mean QTc interval at baseline was longer for women than for men (mean +/- SD; 407 +/- 7 versus 395 +/- 9 ms, P < .05). The slope of the relationship between change in the QTc interval (delta QTc) from baseline to the serum concentration of quinidine was 44% greater for women than for men (mean +/- SE; 42.2 +/- 3.4 versus 29.3 +/- 2.6 ms/microg per mL, P < .001). These results were not influenced by analysis of 3-hydroxyquinidine, free concentrations of quinidine and 3-hydroxyquinidine, or the JT interval. CONCLUSIONS: Quinidine causes greater QT prolongation in women than in men at equivalent serum concentrations. This difference may contribute to the greater incidence of drug induced torsades de pointes observed in women taking quinidine and has implications for other cardiac and noncardiac drugs that prolong the QTc interval. Adjustment of dosages based on body size alone are unlikely to substantially reduce the increased risk of torsades de pointes in women. PMID- 10801252 TI - Study of the analgesic effect of lanepitant in patients with osteoarthritis pain. AB - OBJECTIVE: Lanepitant selectively blocks substance P binding to the neurokinin-1 receptor, preventing neurogenic inflammation and pain transmission. Substance P is present in synovial fluid and in excess in cerebral spinal fluid. We investigated the effect of lanepitant on pain caused by osteoarthritis to evaluate the role of neurokinin-1 blockade. METHODS: Outpatients (n = 214) with moderate to severe lower-limb osteoarthritis pain were treated for 3 weeks in a parallel, randomized double-blind study with initial doses of 20, 60, 200, or 600 mg lanepitant, 375 mg naproxen, or placebo, followed by 10, 30, 100, or 300 mg lanepitant twice a day, 375 mg naproxen twice a day, or placebo twice a day in the multiple-dose period. Pain intensity, pain relief, patient global impression, and adjunctive analgesic use were compared across treatments. Safety was evaluated with adverse events, vital signs, and laboratory assessments. RESULTS: There was no statistically significant difference in efficacy or safety across treatments for the initial dose assessment. After 1 week of therapy, naproxen was statistically significantly (P < .05) better than placebo and lanepitant in reducing average pain. During the second and third weeks of therapy, patients receiving naproxen continued to have statistically significantly (P < .05) less pain than those receiving placebo or lanepitant despite using significantly less adjunctive analgesic medication. There were no statistically significant differences in rates of discontinuation across treatments. Lanepitant treatment was associated with diarrhea, whereas naproxen treatment was associated with gastric discomfort. There were no clinically relevant changes in vital signs or laboratory analytes for any of the treatments. CONCLUSION: Lanepitant was ineffective in relieving osteoarthritis pain, possibly because neurokinin-1 binding of substance P does not play a significant role in osteoarthritis pain or because lanepitant fails to adequately penetrate the blood-brain barrier to affect central pain perception. PMID- 10801253 TI - Effect of fluvastatin on proteinuria in patients with immunoglobulin A nephropathy. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are established drugs for the treatment of hypercholesterolemia, but several studies have shown that benefits obtained with these drugs are not causally related only to regression of cholesterol lowering. Moreover, in experimental models of progressive renal disease, statins have reduced the extent of glomerulosclerosis. This study evaluated the antiproteinuric effect of a daily dose of 40 mg fluvastatin for 6 months in moderately proteinuric patients with immunoglobulin A nephropathy, stable renal function, and no indicators of poor long-term prognosis. The effects of therapy were evaluated on the basis of 24-hour proteinuria (total proteinuria and albuminuria), albuminemia, creatinine clearance, cholesterol, and triglyceride values. Renal function remained stable in all patients. A significant decrease in proteinuria was observed after 6 months of therapy and persisted for all the observations. An increase in serum albumin was observed after 6 months of therapy. This study suggests that there is an antiproteinuric effect of HMG-CoA reductase inhibitors in moderately proteinuric patients with immunoglobulin A nephropathy. PMID- 10801254 TI - Combined glutathione-S-transferase M1 and T1 genetic polymorphism and tacrine hepatotoxicity. AB - BACKGROUND: Glutathione conjugation of tacrine reactive metabolites depends in part on the activity of glutathione-S-transferases (GST), of which two isozymes (GST M1 and GST T1) are polymorphically expressed. OBJECTIVE AND METHODS: To determine whether GST M1, GST T1, and the combined GST M1 and GST T1 null genotypes predict individual susceptibility to tacrine hepatotoxicity, 141 patients with mild to moderate Alzheimer's disease treated with tacrine were genotyped. RESULTS: During the treatment period, 52 patients had elevated alanine aminotransferase (ALT) levels at least three times the upper limit of normal, whereas 89 patients had normal ALT values (< or = upper limit of normal). Both groups were comparable in demographic and clinical characteristics. Twenty-eight patients were found to be GST T1-negative (20%; with a 95% confidence interval [95% CI] from 13% to 27%), and 68 patients (48%; 95% CI from 40% to 57%) were GST M1-negative. The combined GST M1-T1 null genotype was observed in 18 patients (13%; 95% CI from 7% to 18%) of whom 13 had an elevated plasma ALT at least three times the upper limit of normal during the study period. Although the cumulative percentage of elevated plasma ALT tended to be higher in the GST M1 null genotype, neither GST M1 nor GST T1 alone could predict individual susceptibility to tacrine hepatotoxicity. Multivariate Cox hazards model showed that the association of the GST M1-T1 null genotype was an independent risk factor of hepatotoxicity. CONCLUSIONS: The presence of combined alleles M1 and T1 deficiencies in glutathione-S-transferase genes increases the susceptibility to tacrine hepatotoxicity. PMID- 10801255 TI - Growth in use of statins after trials is not targeted to most appropriate patients. AB - OBJECTIVE: To determine whether growth in the use of lipid-lowering drugs after publication of studies in the primary and secondary prevention of coronary heart disease is in the population in which benefit was established, particularly middle-aged men. METHODS: We performed a series of pharmacoepidemiologic surveys of community prescribing in Ireland over 4 years. RESULTS: Nationally, the use of lipid-lowering drugs (92% statins) increased approximately fourfold from 1994 to 1998. In the Eastern Health Board region, the number of monthly recipients increased from 447 in April 1994 to 3530 in March 1998. Although use increased steadily after publication of Scandinavian Simvastatin Survival Study (4S) and West of Scotland Coronary Prevention Study (WOSCOPS) in 1994 and 1995, respectively, this occurred to a greater extent in women. However, after the Cholesterol and Recurrent Events (CARE) study in 1996 and subsequent recommendations that targeted statin use, particularly in men from 35 to 69 years old, there was a relatively greater increase in that population but, at 2.3%, it was well short of the target population of 5.8%. More women than men older than 65 years are receiving statins. The 10-mg dosage (a fourth or half that used in studies) is the most frequently dispensed. CONCLUSION: The use of statins, although rising rapidly, is below targets and was initially not directed at the population likely to benefit most or in the recommended dosage. Consequently, the benefits projected from clinical trials may not be seen in clinical practice. PMID- 10801256 TI - Publishing in the journals of the APS: why are authors charged fees? American Physiological Society. PMID- 10801257 TI - Lessons from genetically engineered animal models XI. Novel mouse models to study pathogenic mechanisms of Crohn's disease. AB - Crohn's Disease (CD) affects more than 500,000 individuals in the United States and represents the second most common chronic inflammatory disorder after rheumatoid arthritis. Although major advances have been made in defining the basic mechanisms underlying chronic intestinal inflammation, the precise etiopathogenesis of CD remains unknown. We have recently characterized two novel mouse models of enteritis that express a CD-like phenotype, namely the TNF DeltaARE model of tumor necrosis factor (TNF) overexpression and the SAMP1/Yit model of spontaneous ileitis. The unique feature of these models is that they closely resemble CD for location and histopathology. These genetically manipulated new models of intestinal inflammation offer a powerful tool to investigate potential causes of human disease and may allow the development of novel disease-modifying therapeutic modalities for the treatment of CD. PMID- 10801258 TI - Pathobiology of visceral pain: molecular mechanisms and therapeutic implications. III. Visceral afferent pathways: a source of new therapeutic targets for abdominal pain. AB - Visceral pain is the major cause of consulting in gastroenterology and the principal symptom of functional bowel disorders. This symptom is often associated with gut hypersensitivity to distension. The use of animal models has recently permitted the identification of some mediators supposed to play a pivotal role in the genesis of visceral hypersensitivity. Serotonin, through different receptor subtypes, as well as kinins and calcitonin gene-related peptide, are known to be involved, but other putative transmitters arise and are new potential targets for the development of efficacious treatments. This themes article addresses both physiological and preclinical issues of interest for the selection of active new drugs in regard to the clinical pharmacology of visceral pain. PMID- 10801259 TI - Effects of enteral glutamine on gut mucosal protein synthesis in healthy humans receiving glucocorticoids. AB - In hypercatabolic patients, the beneficial effects of glutamine on gut mucosa could be partly due to a stimulation of protein synthesis. The fractional synthesis rate (FSR) of gut mucosal protein was measured in four groups of healthy volunteers treated with glucocorticoids for 2 days. Two groups were studied in the postabsorptive state while receiving glutamine or a nitrogen equivalent (control) and two groups in the fed state with or without glutamine, using a 5-h intravenous infusion of [(13)C]leucine, [(2)H(5)]phenylalanine, and cortisone. After nutrient and tracer infusion, duodenal biopsies were taken. In the postabsorptive state, FSR of gut mucosal protein were 87 and 76%/day in the control group and 130% (P = 0.058 vs. control) and 104% (P = 0.17 vs. control)/day in the glutamine group, with leucine and phenylalanine as tracers, respectively. During feeding, FSR did not increase and no significant difference was observed between glutamine and control groups. Overall, FSR of the four groups were two- to threefold higher than those obtained previously in healthy humans, suggesting that glucocorticoids may increase gut mucosal protein synthesis. However, in this situation, a moderate enteral glutamine supply failed to demonstrate a significant effect on gut mucosal protein synthesis in the postabsorptive state and during feeding. PMID- 10801260 TI - Effect of metformin on the vascular and glucose metabolic actions of insulin in hypertensive rats. AB - We investigated the long-term effect of metformin treatment on blood pressure, insulin sensitivity, and vascular responses to insulin in conscious spontaneously hypertensive rats (SHR). The rats were instrumented with intravascular catheters and pulsed Doppler flow probes to measure blood pressure, heart rate, and blood flow. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Two groups of SHR received metformin (100 or 300 mg x kg(-1) x day( 1)) for 3 wk while another group of SHR and a group of Wistar Kyoto (WKY) rats were left untreated. We found that vasodilation of skeletal muscle and renal vasculatures by insulin is impaired in SHR. Moreover, a reduced insulin sensitivity was detected in vivo and in vitro in isolated soleus and extensor digitorum longus muscles from SHR compared with WKY rats. Three weeks of treatment with metformin improves the whole-body insulin-mediated glucose disposal in SHR but has no blood pressure-lowering effect and no influence on vascular responses to insulin (4 mU x kg(-1) x min(-1)). An improvement in insulin-mediated glucose transport activity was detected in isolated muscles from metformin-treated SHR, but in the absence of insulin no changes in basal glucose transport activity were observed. It is suggested that part of the beneficial effect of metformin on insulin resistance results from a potentiation of the hormone-stimulating effect on glucose transport in peripheral tissues (mainly skeletal muscle). The results argue against a significant antihypertensive or vascular effect of metformin in SHR. PMID- 10801261 TI - Role of caspases and NF-kappaB signaling in hydrogen peroxide- and superoxide induced hepatocyte apoptosis. AB - Reactive oxygen intermediates (ROI) have been implicated as mediators of hepatocyte death resulting from a variety of forms of liver injury. To delineate the mechanisms that underlie ROI-induced apoptosis, the roles of caspase activation and nuclear factor-kappaB (NF-kappaB) signaling were determined in the rat hepatocyte cell line RALA255-10G after treatment with H(2)O(2) or the superoxide generator menadione. By 8 h, H(2)O(2) and menadione caused 26% and 33% cell death, respectively. Death from both ROI occurred by apoptosis as indicated by morphology under fluorescence microscopy, the induction of caspase activation and DNA fragmentation, and the cleavage of poly(ADP-ribose) polymerase. Despite the presence of caspase activation in both forms of apoptosis, caspase inhibition blocked H(2)O(2)- but not menadione-induced apoptosis. In contrast, inhibition of NF-kappaB activation decreased cell death from both ROI. Different ROI, therefore, induce distinct apoptotic pathways in RALA hepatocytes that are both caspase dependent and independent. In contrast to the known protective effect of NF-kappaB activation in tumor necrosis factor-alpha-induced hepatocyte apoptosis, NF-kappaB promotes hepatocellular death from ROI in these cells. PMID- 10801262 TI - Differential jejunal and colonic adaptation due to resection and IGF-I in parenterally fed rats. AB - Patients with severe short-bowel syndrome (SBS) often require long-term total parenteral nutrition (TPN) to maintain their nutritional status because of limited intestinal adaptation. Growth factors, including insulin-like growth factor I (IGF-I), are under investigation to promote intestinal adaptation and tolerance to oral feeding. We investigated structural and functional adaptation of the jejunum and colon in four groups of rats maintained with TPN for 7 days after a 60% jejunoileal resection and cecectomy or sham surgery and treatment with IGF-I or vehicle. Resection alone did not stimulate jejunal growth. IGF-I significantly increased jejunal mucosal mass, enterocyte proliferation, and migration rates. IGF-I decreased jejunal sucrase specific activity and reduced active ion transport and ionic permeability; resection alone had no effect. In contrast, resection significantly increased colonic mass and crypt depth but had no effect on active ion transport or ionic permeability. IGF-I had minimal effects on colonic structure. IGF-I but not resection stimulates jejunal adaptation, whereas resection but not IGF-I stimulates colonic growth in rats subjected to a model for human SBS. IGF-I treatment may improve intestinal adaptation in humans with SBS. PMID- 10801263 TI - Endogenous IGF-I regulates IGF binding protein production in human intestinal smooth muscle cells. AB - Human intestinal smooth muscle in culture produces insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-3, IGFBP-4, and IGFBP-5, which modulate the effects of IGF-I. This study examined the regulation of IGFBP production by endogenous IGF-I. R3-IGF-I, an agonist unaffected by IGFBPs, elicited concentration-dependent increase in growth, measured by [(3)H]thymidine incorporation, and production of IGFBP-3, IGFBP-4, and IGFBP-5, measured by Western blot. Antagonists of the IGF-I receptor, IGF-I Analog or monoclonal antibody 1H7, elicited concentration-dependent inhibition of growth and decrease in IGFBP-3, IGFBP-4, and IGFBP-5 production, implying that endogenous IGF-I stimulated growth and IGFBP production. R3-IGF-I-induced increase in IGFBP-3, IGFBP-4, and IGFBP-5 production was partially inhibited by a mitogen-activated protein (MAP) kinase or a phosphatidylinositol-3-kinase (PI 3-kinase) inhibitor and abolished by the combination. We conclude that endogenous IGF-I stimulates growth and IGFBP-3, IGFBP-4, and IGFBP-5 production in human intestinal smooth muscle cells. Regulation of IGFBP production by IGF-I is mediated by activation of distinct MAP kinase and PI 3-kinase pathways, the same pathways through which IGF-I stimulates growth. PMID- 10801264 TI - Early aldosterone effect in distal colon by transcriptional regulation of ENaC subunits. AB - Aldosterone-induced sodium absorption is mediated by the epithelial Na(+) channel (ENaC). It is thought that the "early effect" is not based on genomic regulation of ENaC expression, because ENaC subunit transcription was reported to start later than Na(+) transport. We investigated electrogenic Na(+) absorption (J(Na)) and, in identical tissues, mRNA expression of ENaC subunits in early (EDC) and late (LDC) distal colon of the rat. In both segments, 8-h in vitro incubation with 3 nM aldosterone enhanced expression of beta- and gamma-ENaC mRNA and induced J(Na). J(Na) was 10 times higher in LDC than in EDC. alpha-ENaC mRNA was unchanged in EDC, whereas it decreased in LDC. In LDC, beta- and gamma-ENaC mRNA was induced 1 h after aldosterone addition, whereas J(Na) became apparent >1 h later. Downregulation of alpha-ENaC mRNA did not take part in acute regulation because it started after a lag time of 3 h. Time correlation of beta- and gamma ENaC induction and J(Na) stimulation suggests that the early aldosterone effect on Na(+) absorption in distal colon is caused by transcriptional upregulation of beta- and gamma-ENaC expression. PMID- 10801265 TI - Effect of exogenous ATP on canine jejunal smooth muscle. AB - Intracellular recordings were made from the circular smooth muscle cells of the canine jejunum to study the effect of exogenous ATP and to compare the ATP response to the nonadrenergic, noncholinergic (NANC) inhibitory junction potential (IJP) evoked by electrical field stimulation (EFS). Under NANC conditions, exogenous ATP evoked a transient hyperpolarization (6.5 +/- 0.6 mV) and EFS evoked a NANC IJP (17 +/- 0.4 mV). Omega-conotoxin GVIA (100 nM) and a low-Ca(2+), high-Mg(2+) solution abolished the NANC IJP but had no effect on the ATP-evoked hyperpolarization. The ATP-evoked hyperpolarization and the NANC IJP were abolished by apamin (1 microM) and N(G)-nitro-L-arginine (100 microM). Oxyhemoglobin (5 microM) partially (38.8 +/- 5.5%) reduced the amplitude of the NANC IJP but had no effect on the ATP-evoked hyperpolarization. Neither the NANC IJP nor the ATP-evoked hyperpolarization was affected by P2 receptor antagonists or agonists, including suramin, reactive blue 2, 1-(N, O-bis-[5 isoquinolinesulfonyl]-N-methyl-L-tyrosyl)-4-phenylpiperazine , pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid, alpha, beta-methylene ATP, 2 methylthioadenosine 5'-triphosphate tetrasodium salt, and adenosine 5'-O-2 thiodiphosphate. The data suggest that ATP evoked an apamin-sensitive hyperpolarization in circular smooth muscle cells of the canine jejunum via local production of NO in a postsynaptic target cell. PMID- 10801266 TI - Cytokine and endothelial cell adhesion molecule expression in interleukin-10 deficient mice. AB - The objectives of this study were to quantify cytokine mRNA levels and endothelial cell adhesion molecule message and protein expression in healthy wild type and interleukin-10-deficient (IL-10(-/-)) mice that develop spontaneous and chronic colitis. We found that colonic message levels of IL-1, IL-6, tumor necrosis factor-alpha, interferon-gamma, lymphotoxin-beta, and transforming growth factor-beta were elevated in colitic mice 10- to 35-fold compared with their healthy wild-type controls. In addition, colonic message levels of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) were found to be increased 10-, 5-, and 23-fold, respectively, in colitic IL-10(-/-) mice compared with their wild-type controls. Immunoradiolabeling as well as immunohistochemistry revealed large and significant increases in vascular surface expression of colonic ICAM-1, VCAM-1, and MAdCAM-1 in the mucosa as well as the submucosa of the colons of colitic mice. These data are consistent with the hypothesis that deletion of IL-10 results in the sustained production of proinflammatory cytokines, leading to the upregulation of adhesion molecules and infiltration of mononuclear and polymorphonuclear leukocytes into the cecal and colonic interstitium. PMID- 10801267 TI - Enteric locus of action of prokinetics: ABT-229, motilin, and erythromycin. AB - We investigated the in vivo and in vitro locus of actions of prokinetics: motilin, erythromycin, and ABT-229. The test substances were infused close intra arterially in short segments of the jejunum in the intact conscious state. Each prokinetic acted on a presynaptic neuron and utilized at least one nicotinic synapse to stimulate circular muscle contractions. The final neurotransmitter at the neuroeffector junction was ACh. Motilin and erythromycin, but not ABT-229, also released nitric oxide. Each prokinetic utilized somewhat different subtypes of muscarinic, serotonergic, tachykininergic, and histaminergic receptors, except for the M(3) receptor, which was common to all of them. In contrast, none of the prokinetics stimulated contractions in mucosa-free or mucosa-attached muscle strips, or rings, even though methacholine or electrical field stimulation induced phasic contractions in all of them. The prokinetics also did not release ACh in longitudinal muscle-myenteric plexus preparations. Each prokinetic, however, decreased the length of enzymatically dispersed single cells. In conclusion, each prokinetic may act on a different subset of presynaptic neurons that converge on the postsynaptic cholinergic and nonadrenergic noncholinergic motoneurons. The presynaptic neurons may be impaired in the muscle bath environment. PMID- 10801268 TI - Murine colonic mucosa hyperproliferation. I. Elevated CFTR expression and enhanced cAMP-dependent Cl(-) secretion. AB - Fluid transport in the large intestine is mediated by the cystic fibrosis gene product and cAMP-dependent anion channel cystic fibrosis transmembrane conductance regulator (CFTR). cAMP-mediated Cl(-) secretion by gastrointestinal cell lines in vitro has been positively correlated with the insertion of CFTR into the apical membrane of differentiated senescent colonocytes and negatively correlated with the failure of CFTR to insert into the plasma membrane of their undifferentiated proliferating counterparts. In native tissues, this relationship remains unresolved. We demonstrate, in a transmissible murine colonic hyperplasia (TMCH) model, that (8-fold) colonocyte proliferation was accompanied by increased cellular CFTR mRNA and protein expression (8.3- and 2.4-fold, respectively) and enhanced mucosal cAMP-dependent Cl(-) secretion (2. 3-fold). By immunofluorescence microscopy, cellular CFTR expression was restricted to the apical pole of cells at the base of the epithelial crypt. In contrast, increased cellular proliferation in vivo led to increases in both the cellular level and the total number of cells expressing this anion channel, with cellular CFTR staining extending into the crypt neck region. Hyperproliferating colonocytes accumulated large amounts of CFTR in apically oriented subcellular perinuclear compartments. This novel mode of CFTR regulation may explain why high endogenous levels of cellular CFTR mRNA and protein within the TMCH epithelium were not matched with larger increases in transmucosal CFTR Cl(-) current. PMID- 10801269 TI - Murine colonic mucosa hyperproliferation. II. PKC-beta activation and cPKC mediated cellular CFTR overexpression. AB - In the companion article (Umar S, Scott J, Sellin JH, Dubinsky WP, and Morris AP, Am J Physiol Gastrointest Liver Physiol 278: 753-764, 2000), we have shown that transmissible murine colonic hyperplasia (TMCH) increased cellular cystic fibrosis transmembrane conductance regulator (CFTR) mRNA and protein expression, relocalized CFTR within colonocytes, and enhanced mucosal cAMP-dependent Cl(-) secretion. We show here that these changes were dependent on elevated cellular levels of membrane-bound Ca(2+)- and diacylglycerol-sensitive protein kinase C (PKC) activity (12-fold), induced by selective (3- to 4-fold) rises in conventional PKC (cPKC) isoform expression and membrane translocation. Three cPKC isoforms were detected in isolated crypts: alpha, beta1, and beta2. cPKC-beta1 rises preceded and those of cPKC-alpha and cPKC-beta2 paralleled cellular hyperproliferation and its effects on CFTR expression and cAMP-dependent Cl(-) current secretion. Only cPKC-beta1 and cPKC-beta2 were membrane translocated during TMCH. Furthermore, only cPKC-beta1 trafficked to the nucleus, whereas cPKC beta2 remained partitioned among cytosolic, membrane, and cytoskeletal subcellular fractions. Modest increases in novel PKC-epsilon (nPKC-epsilon) expression and subcellular membrane partitioning were recorded during TMCH, but no changes were seen for PKC-delta or -eta. No nPKC isoform nuclear partitioning was detected. The orally bioactive cPKC inhibitor Ro-32-0432 reversed both TMCH and elevated cellular CFTR mRNA levels, whereas a pharmacologically inert analog (Ro-31-6045) failed to inhibit either response. On the basis of these facts, we present a new hypothesis whereby PKC-dependent cellular proliferation promotes endogenous cellular CFTR levels. PKC-beta1 was identified as a candidate regulatory PKC isoform. PMID- 10801270 TI - Uptake of lactate by the liver: effect of red blood cell carriage. AB - Multiple-indicator dilution experiments with labeled lactate were performed in the livers of anesthetized dogs. A mixture of (51)Cr-labeled erythrocytes, [(3)H]sucrose, and L-[1-(14)C]lactate or a mixture of (51)Cr-labeled erythrocytes, [(14)C]sucrose, and L-[2-(3)H]lactate was injected into the portal vein, and samples were obtained from the hepatic vein. Data were evaluated using a model comprising flow along sinusoids, exchange of lactate between plasma and erythrocytes and between plasma and hepatocytes, and, in the case of L-[1 (14)C]lactate, metabolism to H[(14)C]O(-)(3) within hepatocytes. The coefficient for lactate efflux from erythrocytes was 0.62 +/- 0.24 s(-1), and those for influx into and efflux from hepatocytes were 0.44 +/- 0.13 and 0.14 +/- 0.07 s( 1), respectively. The influx permeability-surface area product of the hepatocyte membrane for lactate (P(in)S, in ml x s(-1) x g(-1)) varied with total flow rate (F, in ml s(-1) x g(-1)) according to P(in)S = (3.1 +/- 0.5)F + (0.021 +/- 0.014). Lactate in plasma, erythrocytes, and hepatocytes was close to equilibrium, whereas lactate metabolism was rate limiting. PMID- 10801271 TI - Effect of cholera toxin on glutamine metabolism and transport in rabbit ileum. AB - The aim of the present study was to evaluate the effect of cholera toxin on energy balance from intestinal glutamine metabolism and oxidation, glutamine dependent sodium absorption, and cholera toxin-dependent ion flux. Cholera toxin stimulated sodium and L-glutamine ileal transport and metabolism were studied in Ussing chambers. Glutamine (10 mM) transport and metabolism were simultaneously studied using (14)C flux and HPLC. In the same tissues, the flux of each amino acid was studied by HPLC, and glutamine metabolism and oxidation were studied by the determination of amino acid specific activity and (14)CO(2) production. In control tissues, glutamine stimulated sodium absorption and was mainly oxidized. The transepithelial flux of intact glutamine represented 45% of glutamine flux across the luminal membrane. The other metabolites were glutamate and, to a lesser degree, citrulline, ornithine, and proline. Cholera toxin did not alter glutamine-stimulated sodium absorption, glutamine oxidation, transport, and metabolism. In conclusion, the present results indicate that cholera toxin does not alter glutamine intestinal function and metabolism. In addition, approximately 95% of the energy provided by glutamine oxidation remains available to the enterocyte. PMID- 10801272 TI - Dextran sulfate sodium-induced murine colitis activates NF-kappaB and increases galanin-1 receptor expression. AB - Galanin is widely distributed in enteric nerve terminals and acts to modulate intestinal motility by altering smooth muscle contraction. This ligand causes Cl( ) secretion when colonic epithelial cells express the galanin-1 receptor (Gal1-R) subtype. Because Gal1-R expression by colonic epithelia is upregulated by the transcription factor nuclear factor-kappaB (NF-kappaB), increasingly appreciated as critical in the pathophysiology of inflammatory bowel disease, we wondered whether the diarrhea associated with this condition could be due to NF-kappaB mediated increases in Gal1-R expression. To test this hypothesis, we provided oral dextran sulfate sodium (DSS) to C57BL/6J mice. Although Gal1-R are not normally expressed by epithelial cells lining the mouse colon, DSS treatment resulted in increased NF-kappaB activation and Gal1-R expression. Whereas galanin had no effect on murine colonic tissues studied ex vivo, it progressively increased short-circuit current and colonic fluid secretion in DSS-treated mice. Concomitant parenteral administration of the NF-kappaB inhibitor dexamethasone attenuated the activation of this transcription factor by DSS, inhibiting the increase in Gal1-R expression. Although Gal1-R-specific antagonists do not exist, intracolonic administration of commercially available galanin antibody diminished the DSS-induced increase in colonic fluid accumulation. Overall, these data demonstrate that a significant component of the excessive fluid secretion observed in DSS-treated mice is due to increased Gal1-R expression. PMID- 10801273 TI - Aging alters gastric mucosal responses to epidermal growth factor and transforming growth factor-alpha. AB - Administration of pharmacological doses of epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha) in young rats stimulates gastric mucosal proliferation, but, in aged rats, the same treatment inhibits proliferation. This may be due to enhanced ligand-induced internalization of EGF receptor (EGFR). In support of this, we demonstrated that although a single injection of EGF (10 microg/kg) or TGF-alpha (5 microg/kg) in young (4-6 mo old) rats greatly increased membrane-associated EGFR tyrosine kinase activity, the same treatment slightly inhibited the enzyme activity in aged (24 mo old) rats. This treatment also produced a greater abundance of punctate cytoplasmic EGFR staining in gastric epithelium of aged rats, consistent with EGFR internalization. In vitro analyses demonstrated that exposure of isolated gastric mucosal cells from aged but not young rats to 100 pM TGF-alpha resulted in marked increases in intracellular EGFR tyrosine kinase activity and that induction of EGFR tyrosine kinase activity in mucosal membranes from aged rats occurred at doses 1,000-fold less than those required in young rats. Our data suggest that aging enhances sensitivity of the gastric mucosa to EGFR ligands. This may partly explain EGFR-mediated inhibition of gastric mucosal proliferation in aged rats. PMID- 10801274 TI - Escherichia coli Shiga toxins induce apoptosis in epithelial cells that is regulated by the Bcl-2 family. AB - Human intestinal cells lack globotriaosylceramide (Gb(3)), the receptor for Shiga toxin-1 (Stx1) and Shiga toxin-2 (Stx2). Therefore, the role of these toxins in mediating intestinal disease during infection with Shiga toxin-producing Escherichia coli is unclear. The aims of this study were to determine whether Stx1 and Stx2 induce apoptosis in epithelial cells expressing (HEp-2, Caco-2) or lacking (T84) Gb(3) and to characterize the role of the Bcl-2 family. Stx1 (12.5 ng/ml) induced apoptosis in both HEp-2 (21.9 +/- 7.9% vs. 0.8 +/- 0.3%, P = 0.01) and Caco-2 (10.1 +/- 1.2% vs. 3.1 +/- 0.4%, P = 0.006) cells but not in Gb(3) deficient T84 cells. Toxin-mediated apoptosis of HEp-2 cells was associated with enhanced expression of the proapoptotic protein Bax. Inhibition of caspase activation prevented toxin-stimulated apoptosis. In addition, overexpression of Bcl-2 by transient transfection blocked Stx1-stimulated cell death. These findings indicate that Shiga toxins produced by E. coli signal Gb(3)-expressing epithelial cells to undergo apoptosis in association with enhanced Bax expression, thereby resulting in activation of the caspase cascade. PMID- 10801275 TI - Cyclooxygenase-1 and an alternatively spliced mRNA in the rat stomach: effects of aging and ulcers. AB - Prostaglandins play a critical role in gastric mucosal cytoprotection and decrease progressively with age. Cyclooxygenase (COX), the rate-limiting enzyme for prostaglandin synthesis, exists in two isoforms, COX-1 and COX-2. The rat COX 1 gene expresses an alternatively spliced mRNA COX-1 splice variant (SV) that may, at best, code for a truncated COX-1 protein. With the use of competitive PCR, we determined whether COX gene expression was altered in the stomach with increasing age and after gastric ulcer induction. COX-1 mRNA was significantly reduced in the aged, and COX-1SV mRNA was significantly higher in the adults compared with the young and aged stomach. Levels of COX-1 and COX-2 were similarly expressed in the normal stomach. In acute gastric ulcers, only COX-2 mRNA levels were significantly elevated. When ulcers were undergoing healing and repair, COX-1 and COX-2 mRNA levels were significantly elevated. Age-related changes in COX-1 and COX-1SV but not COX-2 mRNA may alter gastric mucosal cytoprotection. Furthermore, COX-1 and COX-2 may both contribute to the healing of a gastric ulcer. PMID- 10801276 TI - Publishing in the journals of the APS: why are authors charged fees? PMID- 10801277 TI - Production and functional roles of nitric oxide in the proximal tubule. AB - A significant role for nitric oxide (NO) in proximal tubule physiology and pathophysiology has been revealed by a series of in vivo and in vitro studies. Whether the proximal tubule produces NO under basal conditions is still controversial; however, evidence suggests that the proximal tubule is constantly exposed to NO that might include NO from nonproximal tubule sources. When challenged with a variety of stimuli, including hypoxia, the proximal tubule is able to produce large quantities of NO. In vivo studies generally indicate that NO inhibits fluid and sodium reabsorption by the proximal tubule. However, the final effect of NO on proximal tubular reabsorption appears to depend on the concentration of NO and involve interaction with other regulatory mechanisms. NO regulates Na(+)-K(+)-ATPase, Na(+)/H(+) exchangers, and paracellular permeability of proximal tubular cells, which may contribute to its effect on proximal tubular transport. Enhanced production of NO, perhaps depending on macrophage type inducible NO synthase, participates in hypoxic/ischemic proximal tubular injury. In conclusion, NO plays a fundamental role in both physiology and pathophysiology of the proximal tubule. PMID- 10801278 TI - Angiotensin stimulates respiration in spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) have an activated brain angiotensin system. We hypothesized 1) that ventilation (V) would be greater in conscious SHR than in control Wistar-Kyoto (WKY) rats and 2) that intravenous infusion of the ANG II receptor blocker saralasin would depress respiration in SHR, but not in WKY. Respiration and oxygen consumption (VO(2)) were measured in conscious aged matched groups (n = 16) of adult female SHR and WKY. For protocol 1, rats were habituated to a plethysmograph and measurements obtained over 60-75 min. After installation of chronic intravenous catheters, protocol 2 consisted of 30 min of saline infusion ( approximately 14 microliter. kg(-1). min(-1)) followed by 40 min of saralasin (1.3 microgram. kg(-1). min(-1)). V, tidal volume (VT), inspiratory flow [VT/inspiratory time (TI)], breath expiratory time, and VO(2) were higher, and breath TI was lower in "continuously quiet" SHR. In SHR, but not in WKY rats, ANG II-receptor block decreased V, VT, and VT/TI and increased breath TI. During ANG II-receptor block, an average decrease in VO(2) in SHR was not significant. About one-half of the higher V in SHR appears to be accounted for by an ANG II mechanism acting either via peripheral arterial receptors or circumventricular organs. PMID- 10801279 TI - Hepatoportal bumetanide-sensitive K(+)-sensor mechanism controls urinary K(+) excretion. AB - To determine whether a K(+)-sensor mechanism exists in the hepatoportal region, periarterial hepatic afferent nerve activity responses to intraportal injection of KCl were examined in anesthetized rats. Hepatic afferent nerve activity increased in response to intraportal injection in a K(+) concentration-dependent manner, and the increase was attenuated by inhibition of the Na(+)-K(+)-2Cl(-) cotransporter by bumetanide in a dose-dependent manner. These results suggest that a bumetanide-sensitive K(+)-sensor mechanism exists in the hepatoportal region. Stimulation of this mechanism by intraportal KCl infusion elicited an immediate and powerful kaliuresis with no significant change in the plasma K(+) concentration; this was significantly greater than the kaliuresis induced by intravenous KCl infusion and was attenuated by severing the periarterial hepatic nervous plexus. These results indicate that a hepatoportal bumetanide-sensitive K(+)-sensor mechanism senses the portal venous K(+) concentration and that stimulation of this sensor mechanism causes kaliuresis, which is mainly mediated by the periarterial hepatic nervous plexus. PMID- 10801280 TI - Role of endotoxin in the expression of endothelial selectins after cecal ligation and perforation. AB - The objectives of this study were to determine 1) the changes in endothelial cell adhesion molecule expression that occur in a clinically relevant model of sepsis and 2) the dependence of these changes on endotoxin [lipopolysaccharide (LPS)]. The dual radiolabeled monoclonal antibody technique was used to quantify the expression of E- and P-selectin in LPS-sensitive (C3HeB/FeJ) and LPS-insensitive (C3H/HeJ) mice that were subjected to acute peritonitis by cecal ligation and perforation (CLP). At 6 h after CLP, the expression of both E- and P-selectin was increased in the gut (mesentery, pancreas, and small and large bowel) compared with the sham-operated and/or control animals, with a more marked response noted in LPS-insensitive mice. The lung also exhibited an increased P-selectin expression in both mouse strains. An accumulation of granulocytes, assessed using tissue myeloperoxidase activity, was noted in the lung and intestine of LPS sensitive but not LPS-insensitive mice exposed to CLP. These results indicate that the CLP model of sepsis is associated with an upregulation of endothelial selectins in the gut vasculature and that enteric LPS does not contribute to this endothelial cell activation response. PMID- 10801281 TI - Range of entrainment of rat circadian rhythms to sinusoidal light-intensity cycles. AB - The range of entrainment of the circadian behavioral rhythm was compared between two groups of Sprague-Dawley rats (each n = 10) exposed to daily cycles of rectangular light-dark alternation (LD) and sinusoidal fluctuations of light intensity (SINE), respectively. The maximum illuminance (20 lx), the minimum illuminance (0.01 lx), and the total amount of light exposure per cycle were the same under the two lighting conditions. The periods (Ts) of both lighting cycles were lengthened stepwise from 24 through 25, 26, 26.5, 27, 27. 5, and 28 h to 28.5 h in experiment 1 and were shortened stepwise from 24 through 23.5, 23, and 22.5 h to 22 h in experiment 2. Each T cycle lasted for 30 cycles. In experiment 1, 60% of rats under the LD condition entrained up to T = 28.5 h, whereas 50% of rats under the SINE condition entrained up to T = 28.5 h. In experiment 2, no animal under the LD condition entrained to T < 23.5 h, whereas 40% of rats under the SINE condition entrained down to T = 23 h and 20% of rats remained to entrain down to T = 22 h cycles. The phase angle of entrainment was systematically changed, depending on T under both conditions. These results suggest that the lower limit of entrainment is expanded under the SINE condition compared with the LD condition. PMID- 10801282 TI - Central control of cardiac baroreflex responses during peripheral hyperosmolality. AB - Acute increases in peripheral osmolality evoke a pressor response and baroreflex mediated bradycardia. These experiments were designed to determine if the fall in heart rate during peripheral sodium loading is 1) equivalent to bradycardia accompanying phenylephrine (PE) infusion, 2) mediated by the parasympathetic (PSNS) or sympathetic (SNS) nervous system, and 3) controlled by the median preoptic nucleus (MnPO). Male rats received an intravenous infusion of isotonic saline, hypertonic saline (2.5 M NaCl), or PE for 30 min. Blood pressure increased equivalently in the hypertonic NaCl and PE groups. However, heart rate fell more in animals infused with PE. Furthermore, pretreatment with methylatropine to block the PSNS had no effect on bradycardia, whereas blocking SNS influences on cardiac function significantly attenuated the fall in heart rate during peripheral hyperosmolality. Finally, kainic acid administration in the MnPO before testing increased bradycardia observed during hypertonic saline loading. Taken together, these data suggest that acute peripheral hyperosmolality acts at the MnPO to reduce cardiac SNS withdrawal during the pressor response that reduces the associated baroreflex bradycardia. PMID- 10801283 TI - Hepatic glucokinase is induced by dietary carbohydrates in rainbow trout, gilthead seabream, and common carp. AB - Glucokinase (GK) plays a central role in glucose homeostasis in mammals. The absence of an inducible GK has been suggested to explain the poor utilization of dietary carbohydrates in rainbow trout. In this context, we analyzed GK expression in three fish species (rainbow trout, gilthead seabream, and common carp) known to differ in regard to their dietary carbohydrate tolerance. Fish were fed for 10 wk with either a diet containing a high level of digestible starch (>20%) or a diet totally deprived of starch. Our data demonstrate an induction of GK gene expression and GK activity by dietary carbohydrates in all three species. These studies strongly suggest that low dietary carbohydrate utilization in rainbow trout is not due to the absence of inducible hepatic GK as previously suggested. Interestingly, we also observed a significantly lower GK expression in common carp (a glucose-tolerant fish) than in rainbow trout and gilthead seabream, which are generally considered as glucose intolerant. These data suggest that other biochemical mechanisms are implicated in the inability of rainbow trout and gilthead seabream to control blood glucose closely. PMID- 10801284 TI - Prevention of hypoinsulinemia modifies catecholamine effects in fetal sheep. AB - Increased epinephrine (Epi) and norepinephrine (NE) production plays an important role in fetal adaptation to reduced oxygen and/or nutrient availability, inhibiting insulin secretion and slowing growth to support more essential processes. To assess the importance of hypoinsulinemia for the efficacy of catecholamines, normoinsulinemia was restored by intravenous insulin infusion (0.18 mU. kg(-1). min(-1)) during prolonged infusion of either Epi (0.25-0. 35 microgram. kg(-1). min(-1) for 12 days, n = 7) or NE (0.5-0.7 microgram. kg(-1). min(-1) for 7 days, n = 6) into normoxemic fetuses in twin-pregnant ewes, from 125-127 days of gestation. Insulin infusion for 8 days during Epi infusion or for 4 days during NE infusion decreased arterial blood pressure, O(2) content, and plasma glucose, but increased heart rate significantly (all P <0.05), despite continuation of Epi or NE infusion. Cessation of insulin infusion reversed these changes. Estimated growth of fetuses infused with insulin during Epi or NE infusion (55 +/- 13.9 and 83 +/- 15.2 g/day) did not differ significantly from that of untreated controls (72 +/- 15.4 g/day, n = 6). Growth of selected muscles and hindlimb bones was not altered either. Restoration of normoinsulinemia evidently counteracts the redistribution of metabolic activity and decreased anabolism brought about by Epi or NE in the fetus. Inhibition of insulin secretion by Epi and NE, therefore, appears essential for the efficacy of catecholamine action in the fetus. PMID- 10801285 TI - Regulation of protein breakdown and adrenocortical response to stress in birds during migratory flight. AB - During long-term fasting at rest, protein utilization is maintained at low levels until it increases at a threshold adiposity. This study examines 1) whether such a shift in energy substrate use also occurs during endurance exercise while fasting, 2) the role of corticosterone, and 3) the adrenocortical response to an acute stressor. Ten species of migrating birds caught after an endurance flight over at least 500 km were examined. Plasma uric acid and corticosterone levels were low in birds with fat stores >5% of body mass and high in birds with smaller fat stores. Corticosterone levels were very high in birds with no visible fat stores and emaciated breast muscles. Corticosterone levels increased with handling time only in birds with large fat stores. These findings suggest that 1) migrating birds with appreciable fat stores are not stressed by endurance flight, 2) a metabolic shift (increased protein breakdown), regulated by an endocrine shift (medium corticosterone levels), occurs at a threshold adiposity, as observed in birds at rest, 3) adrenocortical response to an acute stressor is inhibited after this shift, and 4) an adrenocortical response typical for an emergency situation (high corticosterone levels) is only reached when muscle protein is dangerously low. PMID- 10801286 TI - An endogenous monocarboxylate transport in Xenopus laevis oocytes. AB - We investigated the existence of an endogenous system for lactate transport in Xenopus laevis oocytes. (36)Cl-uptake studies excluded the involvement of a DIDS sensitive anion antiporter as a possible pathway for lactate movement. L [(14)C]lactate uptake was unaffected by superimposed pH gradients, stimulated by the presence of Na(+) in the incubating solution, and severely reduced by the monocarboxylate transporter inhibitor p-chloromercuribenzenesulphonate (pCMBS). Transport exhibited a broad cation specificity and was cis inhibited by other monocarboxylates, mostly by pyruvate. These results suggest that lactate uptake is mediated mainly by a transporter and that the preferred anion is pyruvate. [(14)C]pyruvate uptake exhibited the same pattern of functional properties evidenced for L-lactate. Kinetic parameters were calculated for both monocarboxylates, and a higher affinity for pyruvate was revealed. Various inhibitors of monocarboxylate transporters reduced significantly pyruvate uptake. These studies demonstrate that Xenopus laevis oocytes possess a monocarboxylate transport system that shares some functional features with the members of the mammalian monocarboxylate cotransporters family, but, in the meanwhile, exhibits some particular properties, mainly concerning cation specificity. PMID- 10801287 TI - Lipopolysaccharide and D-galactosamine-induced hepatic injury is mediated by TNF alpha and not by Fas ligand. AB - Tumor necrosis factor (TNF)-alpha and Fas ligand (FasL) are trimeric proteins that induce apoptosis through similar caspase-dependent pathways. Hepatocytes are particularly sensitive to inflammation-induced programmed cell death, although the contribution of TNF-alpha and/or FasL to this injury response is still unclear. Here, we report that D-galactosamine and lipopolysaccharide-induced liver injury in C57BL/6 mice is associated with increased hepatic expression of both TNF-alpha and FasL mRNA. Pretreatment of mice with a TNF-binding protein improved survival, reduced plasma aspartate aminotransferase concentrations, and attenuated the apoptotic liver injury, as determined histologically and by in situ 3' OH end labeling of fragmented nuclear DNA. In contrast, pretreatment of mice with a murine-soluble Fas fusion protein (Fasfp) had only minimal effect on survival, and apoptotic liver injury was either unaffected or exacerbated depending on the dose of Fasfp employed. Similarly, mice with a spontaneous mutation in FasL (B6Smn.C3H-Fasl(gld) derived from C57BL/6) were equally sensitive to D-galactosamine/lipopolysaccharide-induced shock. We conclude that the shock and apoptotic liver injury after D-galactosamine/lipopolysaccharide treatment are due primarily to TNF-alpha release, whereas increased FasL expression appears to contribute little to the mortality and hepatic injury. PMID- 10801288 TI - LPS-induced liver injury in D-galactosamine-sensitized mice requires secreted TNF alpha and the TNF-p55 receptor. AB - Lipopolysaccharide and D-galactosamine induced lethality and apoptotic liver injury is dependent on endogenously produced tumor necrosis factor (TNF)-alpha. The present study was undertaken to determine whether membrane-associated or secreted TNF-alpha signaling through the p55 or p75 receptor was responsible for survival and hepatic injury after lipopolysaccharide administration in D galactosamine-sensitized mice. Transgenic mice expressing null forms of TNF alpha, the p55 and p75 receptor, and mice expressing only a cell-associated form of TNF-alpha were challenged with 8 mg D-galactosamine and 100 ng lipopolysaccharide. Mortality and apoptotic liver injury were only seen in wild type and p75 knockout mice. p75 Knockout mice had significantly higher concentrations of plasma TNF-alpha than any other experimental group (P 0.05). Selective iNOS inhibition restored the AO's response to vasoconstriction (LPS + AG 1,135 +/- 54 mg, P > 0.05 vs. control and P < 0.05 vs. LPS), but had no effect on the PA (LPS + AG 422 +/- 38 mg, P > 0.05 vs. control and LPS). Western blot and immunohistochemistry revealed increased iNOS expression in the AO after LPS, but iNOS was not detected in the PA. Our results suggest that differential iNOS expression after LPS in systemic and pulmonary vessels contributes to the phenomenon of sepsis/endotoxemia-induced systemic hypotension and pulmonary hypertension. PMID- 10801293 TI - Hyperthermia impairs liver mitochondrial function in vitro. AB - The effects of temperature on the relationships among the rates of pyruvate carboxylation, O(2) uptake (J(o)), oxidative phosphorylation (J(p)), and the free energy of ATP hydrolysis (G(p)) were studied in liver mitochondria isolated from 250-g female rats. Pyruvate carboxylation was evaluated at 37, 40, and 43 degrees C. In disrupted mitochondria, pyruvate carboxylase maximal reaction velocity increased from 37 to 43 degrees C with an apparent Q(10) of 2.25. A reduction in ATP/ADP ratio decreased enzyme activity at all three temperatures. In contrast, in intact mitochondria, increasing temperature failed to increase pyruvate carboxylation (malate + citrate accumulation) but did result in increased J(o) and decreased extramitochondrial G(p). J(p) was studied in respiring mitochondria at 37 and 43 degrees C at various fractions of state 3 respiration, elicited with a glucose + hexokinase ADP-regenerating system. The relationship between J(o) and G(p) was similar at both temperatures. However, hyperthermia (43 degrees C) reduced the J(p)/J(o) ratio, resulting in lower G(p) for a given J(p). Fluorescent measurements of membrane phospholipid polarization revealed a transition in membrane order between 40 and 43 degrees C, a finding consistent with increased membrane proton conductance. It is concluded that hyperthermia augments nonspecific proton leaking across the inner mitochondrial membrane, and the resultant degraded energy state offsets temperature stimulation of pyruvate carboxylase. As a consequence, at high temperatures approaching 43 degrees C, the pyruvate carboxylation rate of intact liver mitochondria may fail to exhibit a Q(10) effect. PMID- 10801294 TI - Explicating hypergonadotropism in postmenopausal women: a statistical model. AB - Neurohormone secretion is viewed here as a variable (unknown) admixture of basal and pulsatile release mechanisms, convolved with individually fitted biexponential elimination kinetics. This construct allows maximum-likelihood estimates of both (regulated and constitutive) components of hormone secretion. Thereby, we infer that a prolonged slow-component half-life of gonadotropin removal and amplified pulsatile (and total) daily luteinizing hormone (LH) secretion rates jointly explicate the postmenopausal elevation in serum LH concentrations without a necessary rise in basal LH secretion rates. This biomathematical formulation should be useful in exploring other neuroregulatory mechanisms that underlie single or dual alterations in the basal versus pulsatile modes of hormone secretion. PMID- 10801295 TI - Enhanced pressor response to carotid occlusion in commNTS-lesioned rats: possible efferent mechanisms. AB - Bilateral common carotid occlusion (BCO) over a period of 60 s in conscious rats produces a biphasic pressor response, consisting of an early (peak) and late (plateau) phase. In this study we investigated 1) the effects of lesions of the commissural nucleus of the solitary tract (commNTS) on the cardiovascular responses produced by BCO in conscious rats and 2) the autonomic and humoral mechanisms activated to produce the pressor response to BCO in sham- and commNTS lesioned rats. Both the peak and plateau of the pressor response produced by BCO increased in commNTS-lesioned rats despite the impairment of chemoreflex responses induced by intravenous potassium cyanide. In sham rats sympathetic blockade with intravenous prazosin and metoprolol, but not vasopressin receptor blockade with the Manning compound, reduced both components of BCO. In commNTS lesioned rats the sympathetic blockade or vasopressin receptor blockade reduced both components of BCO. The results showed 1) the sympathetic nervous system, but not vasopressin, is important for the pressor response to BCO during 60 s in conscious sham rats; 2) in commNTS-lesioned rats, despite chemoreflex impairment, BCO produces an increased pressor response dependent on sympathetic activity associated with vasopressin release; and 3) the increment in the pressor response to BCO in commNTS-lesioned rats seems to depend only on vasopressin secretion. PMID- 10801296 TI - Rhythms in Fos expression in brain areas related to the sleep-wake cycle in the diurnal Arvicanthis niloticus. AB - Most mammals show daily rhythms in sleep and wakefulness controlled by the primary circadian pacemaker, the suprachiasmatic nucleus (SCN). Regardless of whether a species is diurnal or nocturnal, neural activity in the SCN and expression of the immediate-early gene product Fos increases during the light phase of the cycle. This study investigated daily patterns of Fos expression in brain areas outside the SCN in the diurnal rodent Arvicanthis niloticus. We specifically focused on regions related to sleep and arousal in animals kept on a 12:12-h light-dark cycle and killed at 1 and 5 h after both lights-on and lights off. The ventrolateral preoptic area (VLPO), which contained cells immunopositive for galanin, showed a rhythm in Fos expression with a peak at zeitgeber time (ZT) 17 (with lights-on at ZT 0). Fos expression in the paraventricular thalamic nucleus (PVT) increased during the morning (ZT 1) but not the evening activity peak of these animals. No rhythm in Fos expression was found in the centromedial thalamic nucleus (CMT), but Fos expression in the CMT and PVT was positively correlated. A rhythm in Fos expression in the ventral tuberomammillary nucleus (VTM) was 180 degrees out of phase with the rhythm in the VLPO. Furthermore, Fos production in histamine-immunoreactive neurons of the VTM cells increased at the light-dark transitions when A. niloticus show peaks of activity. The difference in the timing of the sleep-wake cycle in diurnal and nocturnal mammals may be due to changes in the daily pattern of activity in brain regions important in sleep and wakefulness such as the VLPO and the VTM. PMID- 10801297 TI - Evidence that platelet-derived growth factor may be a novel endogenous pyrogen in the central nervous system. AB - Platelet-derived growth factor (PDGF) exerts neurotrophic and neuromodulatory actions in the mammalian central nervous system (CNS). Like the cytokines, PDGF primarily signals through tyrosine phosphorylation-dependent pathways that activate multiple intracellular molecules including Janus family kinases. We previously showed that microinjection of PDGF-BB into the lateral ventricle induced a febrile response in rats that was reduced by pretreatment with Win 41662, a potent inhibitor of PDGF receptors (Pela IR, Ferreira MES, Melo MCC, Silva CAA, and Valenzuela CF. Ann NY Acad Sci 856: 289-293, 1998). In this study, we further characterized the role of PDGF-BB in the febrile response in rats. Microinjection of PDGF-BB into the third ventricle produced a dose-dependent increase in colonic temperature that peaked 3-4 h postinjection. Win 41662 attenuated fever induced by intraperitoneal injection of bacterial lipopolysaccharide, suggesting that endogenous PDGF participates in the febrile response to this exogenous pyrogen. Importantly, febrile responses induced by tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 were unchanged by Win 41662. Both indomethacin and dexamethasone blocked the PDGF-BB-induced increase in colonic temperature, and, therefore, we postulate that PDGF-BB may act via prostaglandin- and/or inducible enzyme-dependent pathways. Thus our findings suggest that PDGF-BB is an endogenous CNS mediator of the febrile response in rats. PMID- 10801298 TI - Eccentric ergometry: increases in locomotor muscle size and strength at low training intensities. AB - Lengthening (eccentric) muscle contractions are characterized by several unusual properties that may result in unique skeletal muscle adaptations. In particular, high forces are produced with very little energy demand. Eccentrically trained muscles gain strength, but the specific nature of fiber size and composition is poorly known. This study assesses the structural and functional changes that occur to normal locomotor muscle after chronic eccentric ergometry at training intensities, measured as oxygen uptake, that do not influence the muscle when exercised concentrically. Male subjects trained on either eccentric or concentric cycle ergometers for 8 wk at a training intensity starting at 54% and ending at 65% of their peak heart rates. The isometric leg strength increased significantly in the eccentrically trained group by 36%, as did the cross-sectional area of the muscle fiber by 52%, but the muscle ultrastructure remained unchanged. There were no changes in either fiber size, composition, or isometric strength in the concentrically trained group. The responses of muscle to eccentric training appear to be similar to resistance training. PMID- 10801299 TI - Effects of abdominal or cardiopulmonary sympathetic afferents on upper cervical inspiratory neurons. AB - Responses of upper cervical inspiratory neurons (UCINs) to abdominal visceral or cardiopulmonary sympathetic stimulation were studied using extracellular recordings from 213 UCINs in 54 pentobarbital sodium-anesthetized and paralyzed rats. Phrenic nerve activity was used to assess inspiration. The UCINs discharging during inspiration only were mainly in the C(1) segment, whereas phase-spanning UCINs were mostly in the C(2) segment. Phase-spanning activity was typically retained after overventilation or vagotomy. When greater splanchnic nerve (GSN) or cardiopulmonary sympathetic afferent (CPSA) fibers were electrically stimulated, augmented UCIN activity was observed in 65% of cells responding to CPSA stimulation but in only 17% of cells responding to GSN. Response latencies were 10.7 +/- 0.5 and 20.6 +/- 1.5 (SE) ms, respectively. Many augmented responses to CPSA stimulation (64%) and all augmented responses to GSN stimulation were followed by suppression of UCIN discharge (biphasic response). Phrenic nerve activity was suppressed by both GSN and CPSA stimulation, but with shorter latency for the latter (29 +/- 0.7 vs. 14.0 +/- 0.7 ms). Excitation of UCINs using CPSA stimulation occurs more often and by a more direct pathway than for GSN input. PMID- 10801300 TI - Feeding-entrained circadian rhythms are attenuated by lesions of the parabrachial region in rats. AB - Rats anticipate daily restricted meals with increased approaches to a feeder and an increase in core body temperature. Food anticipatory activity (FAA) is thought to be under the control of a feeding-entrained circadian oscillator. Although numerous forebrain lesions have failed to permanently abolish FAA, the hindbrain has not been investigated. The parabrachial nuclei (PBN) integrate information from visceral and gustatory afferents. This region is also innervated by neurons in the area postrema that have access to the peripheral circulation. Therefore, it is possible that this region plays a role in triggering FAA. In two experiments, a total of 19 rats were given ibotenic acid or electrolytic lesions targeted at the PBN. The PBN-lesioned animals showed a marked attenuation in anticipatory approaches to the food bin relative to sham-operated controls. Some animals did not anticipate the meal at all. In addition, the expected increase in core body temperature was severely attenuated in the PBN-lesioned animals compared with controls. The most likely interpretation of these data is that the PBN serve as a relay for information about the zeitgeber (food in the gut) or as a clock output pathway, but not as the site of the feeding-entrained circadian oscillator. PMID- 10801301 TI - Maturational differences between vascular and bladder smooth muscle during ovine development. AB - Maturation rates of vascular and visceral smooth muscle (SM) during ovine development were compared by quantifying contractile protein, myosin heavy chain (MHC) isoform contents, and contractile properties of aortas and bladders from female fetal (n = 19) and postnatal (n = 21) sheep. Actin, myosin, and protein contents rose progressively throughout development in both tissues (P /= 40 microM). Complementary NMR measurements have shown that neomycin forms both a specific binary complex with RRE and a specific ternary complex with RRE and Rev. NMR data further suggest that neomycin occupies a similar high-affinity binding site in both the binary and ternary complexes, and that this site is located in the lower stem region of RRE. PMID- 10801314 TI - Phosphorothioate substitution can substantially alter RNA conformation. AB - Phosphorothioate substitution-interference experiments, routinely used to stereospecifically identify phosphoryl oxygen sites that participate in RNA ligand binding and RNA-directed catalysis, rest in their interpretation on the untested assumption that substitution does not alter the conformation of the modified molecule from its biologically active state. Using NMR spectroscopy, we have tested this assumption by determining the structural effect of stereospecific phosphorothioate substitution at five positions in an RNA hairpin containing the binding site for bacteriophage MS2 capsid protein. At most sites, substitution has little or no effect, causing minor perturbations in the phosphate backbone and increasing the stacking among nucleotides in the hairpin loop. At one site, however, phosphorothioate substitution causes an unpaired adenine necessary for formation of the capsid protein-RNA complex to loop out of the RNA helix into the major groove. These results indicate that phosphorothioate substitution can substantially alter the conformation of RNA at positions of irregular secondary structure, complicating the use of substitution-interference experiments to study RNA structure and function. PMID- 10801315 TI - Distances between DNA and ATP binding sites in the TyrR-DNA complex. AB - The Escherichia coli regulatory protein TyrR controls the expression of eight transcription units that encode proteins involved in the biosynthesis and transport of aromatic amino acids. It binds to DNA as a homodimer with a subunit molecular mass of 57 640 Da, each of which has a single site for the binding of ATP within a central structural domain. This paper reports distances between four sites on the DNA and the ATP binding site as determined by fluorescence resonance energy transfer. The DNA was a 30mer containing a centrally located binding site for TyrR. Replacement of a thymidine residue with an aminouridine residue at positions -9, -7, -3, and 2 of the palindromic oligonucleotide sequence enabled the placement of a single fluorescein group along the major groove of the DNA. The energy transfer acceptor was ATP labeled with a rhodamine group through positions 2' and 3' of the ribose, positions that are known to cause minimal interference with the binding of ATP to protein. The dissociation constant for the binding of rhodamine-ATP to TyrR was 300 nM as determined by steady-state fluorescence anisotropy titrations. The energy transfer efficiencies were determined by measuring the level of quenching of donor fluorescence on binding rhodamine-ATP to the TyrR-DNA complex. The experimental transfer efficiencies were compared to theoretical values calculated for a model of the DNA-TyrR complex in which the position of the ATP binding site was allowed to vary over the surface of the monomer unit. Theory was written to account for the transfer from one donor to two acceptors, one on each monomer unit of the TyrR dimer. The results indicate that the ATP binding site is about 40-45 A from the nearest point on the DNA and distant from the DNA helix-turn-helix binding domain. The effects of ATP binding of (i) increasing the TyrR binding affinity by a factor of 4-5 and (ii) permitting the binding of the tyrosine corepressor must therefore occur because of a significant allosteric change in the conformation of the protein. PMID- 10801316 TI - Probing the multidomain structure of the type I regulatory subunit of cAMP dependent protein kinase using mutational analysis: role and environment of endogenous tryptophans. AB - The regulatory R-subunit of cAMP-dependent protein kinase (cAPK) is a thermostable multidomain protein. It contains a dimerization domain at the N terminus followed by an inhibitor site that binds the catalytic C-subunit and two tandem cAMP-binding domains (A and B). Two of the three tryptophans in the RIalpha subunit, Trp188 and Trp222, lie in cAMP-binding domain A while Trp260 lies at the junction between domains A and B. The unfolding of wild-type RIalpha (wt-RI), monitored by intrinsic fluorescence, was described previously [Leon, D. A., Dostmann, W. R. G., and Taylor, S. S. (1991) Biochemistry 30, 3035 (1)]. To determine the environment of each tryptophan and the role of the adjacent domain in folding and stabilization of domain A, three point mutations, W188Y, W222Y, and W260Y, were introduced. The secondary structure of wt-RI and the point mutants has been studied by far-UV circular dichroism spectropolarimetry (CD). The CD spectra of wt-RI and the three point mutants are practically identical, and the thermal unfolding behavior is very similar. Intrinsic fluorescence and iodide quenching in the presence of increasing urea established that: (a) Trp222 is the most buried, whereas Trp188 is the most exposed to solvent; (b) Trp260 accounts for the quenching of fluorescence when cAMP is bound; and (c) Trp222 contributes most to the intrinsic fluorescence of the wt-RI-subunit, while Trp188 contributes least. For wt-RI, rR(W188Y), and rR(W260Y), removal of cAMP causes a destabilization, while excess cAMP stabilizes these three proteins. In contrast, rR(W222Y) was not stabilized by excess cAMP. PMID- 10801317 TI - Partially unfolded species populated during equilibrium denaturation of the beta sheet protein Y74W apo-pseudoazurin. AB - Apo-pseudoazurin is a single domain cupredoxin. We have engineered a mutant in which a unique tryptophan replaces the tyrosine residue found in the tyrosine corner of this Greek key protein, a region that has been proposed to have an important role in folding. Equilibrium denaturation of Y74W apo-pseudoazurin demonstrated multistate unfolding in urea (pH 7.0, 0.5 M Na(2)SO(4) at 15 degrees C), in which one or more partially folded species are populated in 4. 3 M urea. Using a variety of biophysical techniques, we show that these species, on average, have lost a substantial portion of the native secondary structure, lack fixed tertiary packing involving tryptophan and tyrosine residues, are less compact than the native state as determined by fluorescence lifetimes and time resolved anisotropy, but retain significant residual structure involving the trytophan residue. Peptides ranging in length from 11 to 30 residues encompassing this region, however, did not contain detectable nonrandom structure, suggesting that long-range interactions are important for stabilizing the equilibrium partially unfolded species in the intact protein. On the basis of these results, we suggest that the equilibrium denaturation of Y74W apo-pseudoazurin generates one or more partially unfolded species that are globally collapsed and retain elements of the native structure involving the newly introduced tryptophan residue. We speculate on the role of such intermediates in the generation of the complex Greek key fold. PMID- 10801318 TI - Redesign of artificial globins: effects of residue replacements at hydrophobic sites on the structural properties. AB - Artificial sequences of the 153 amino acids have been designed to fit the main chain framework of the sperm whale myoglobin (Mb) structure based on a knowledge based 3D-1D compatibility method. The previously designed artificial globin (DG1) folded into a monomeric, compact, highly helical and globular form with overall dimensions similar to those of the target structure, but it lacked structural uniqueness at the side-chain level [Isogai, Y., Ota, M., Fujisawa, T. , Izuno, H., Mukai, M., Nakamura, H., Iizuka, T., and Nishikawa, K. (1999) Biochemistry 38, 7431-7443]. In this study, we redesigned hydrophobic sites of DG1 to improve the structural specificity. Several Leu and Met residues in DG1 were replaced with beta-branched amino acids, Ile and Val, referring to the 3D profile of DG1 to produce three redesigned globins, DG2-4. These residue replacements resulted in no significant changes of their compactness and alpha-helical contents in the absence of denaturant, whereas they significantly affected the dependence of the secondary structure on the concentration of guanidine hydrochloride. The analyses of the denaturation curves revealed higher global stabilities of the designed globins than that of natural apoMb. Among DG1-4, DG3, in which 11 Leu residues of DG1 are replaced with seven Ile and four Val residues, and one Met residue is replaced with Val, displayed the lowest stability but the most cooperative folding-unfolding transition and the most dispersed NMR spectrum with the smallest line width. The present results indicate that the replacements of Leu (Met) with the beta-branched amino acids at appropriate sites reduce the freedom of side-chain conformation and improve the structural specificity at the expense of stability. PMID- 10801319 TI - Crystallographic evidence for Tyr 157 functioning as the active site base in human UDP-galactose 4-epimerase. AB - UDP-galactose 4-epimerase catalyzes the interconversion of UDP-glucose and UDP galactose during normal galactose metabolism. In humans, deficiencies in this enzyme lead to the complex disorder referred to as epimerase-deficiency galactosemia. Here, we describe the high-resolution X-ray crystallographic structures of human epimerase in the resting state (i.e., with bound NAD(+)) and in a ternary complex with bound NADH and UDP-glucose. Those amino acid side chains responsible for anchoring the NAD(+) to the protein include Asp 33, Asn 37, Asp 66, Tyr 157, and Lys 161. The glucosyl group of the substrate is bound to the protein via the side-chain carboxamide groups of Asn 187 and Asn 207. Additionally, O(gamma) of Ser 132 and O(eta) of Tyr 157 lie within 2.4 and 3.1 A, respectively, of the 4'-hydroxyl group of the sugar. Comparison of the polypeptide chains for the resting enzyme and for the protein with bound NADH and UDP-glucose demonstrates that the major conformational changes which occur upon substrate binding are limited primarily to the regions defined by Glu 199 to Asp 240 and Gly 274 to Tyr 308. Additionally, this investigation reveals for the first time that a conserved tyrosine, namely Tyr 157, is in the proper position to interact directly with the 4'-hydroxyl group of the sugar substrate and to thus serve as the active-site base. A low barrier hydrogen bond between the 4' hydroxyl group of the sugar and O(gamma) of Ser 132 facilitates proton transfer from the sugar 4'-hydroxyl group to O(eta) of Tyr 157. PMID- 10801320 TI - Control of the chemical step by leucine-31 of pancreatic phospholipase A2. AB - A well-defined region of pancreatic and other secreted phospholipase A2 (PLA2), which we call the i-face, makes a molecular contact with the interface to facilitate and control the events and processivity of the interfacial catalytic turnover cycles. The structural features of the i-face and its allosteric relationship to the active site remain to be identified. As a part of the calcium binding (26-34) loop, Leu-31 is located on the surface near the substrate binding slot of PLA2. Analysis of the primary rate and equilibrium parameters of the Leu 31 substitution mutants of the pig pancreatic PLA2 shows that the only significant effect of the substitution is to impair the chemical step at the zwitterionic interface in the presence of added NaCl, and only a modest effect is seen on kcat at the anionic interface. Leu-31 substitutions have little effect on the binding of the enzyme to the interface; the affinity for certain substrate mimics is modestly influenced in W3F, L31W double mutant. The fluorescence emission results with the double mutant show that the microenvironment of Trp-31 is qualitatively different at the zwitterionic versus anionic interfaces. At both of the interfaces Trp-31 is not shielded from the bulk aqueous environment as it remains readily accessible to acrylamide and water. The NaCl-induced change in the Trp-31 emission spectrum of the double mutant on the zwitterionic interface is similar to that seen on the binding to the anionic interface. Together, the kinetic and spectroscopic results show that the form of PLA2 at the zwitterionic interface (Ez) is distinguishably different from the catalytically more efficient form at the anionic interface (Ea). This finding provides a structural basis for the two-state model for kcat activation by the anionic interface. In conjunction with earlier results we suggest that neutralization of certain cationic residues of PLA2 exerts a control on the calcium loop through residue 31. PMID- 10801321 TI - The conformation of apolipoprotein A-I in high-density lipoproteins is influenced by core lipid composition and particle size: a surface plasmon resonance study. AB - Plasma high-density lipoproteins (HDL) are a heterogeneous group of particles that vary in size as well as lipid and apoprotein composition. The effect of HDL core lipid composition and particle size on apolipoprotein (apo) A-I structure was studied using surface plasmon resonance (SPR) analysis of the binding of epitope-defined monoclonal antibodies. The association and dissociation rate constants of 12 unique apo A-I-specific monoclonal antibodies for isolated plasma HDL were calculated. In addition, the association rate constants of the antibodies were determined for homogeneous preparations of spherical reconstituted HDL (rHDL) that contained apo A-I as the sole apolipoprotein and differed either in their size or in their core lipid composition. This analysis showed that lipoprotein size affected the conformation of domains dispersed throughout the apo A-I molecule, but the conformation of the central domain between residues 121 and 165 was most consistently modified. In contrast, replacement of core cholesteryl esters with triglyceride in small HDL modified almost the entire molecule, with only two key N-terminal domains of apo A-I being unaffected. This finding suggested that the central and C-terminal domains of apo A-I are in direct contact with rHDL core lipids. This immunochemical analysis has provided valuable insight into how core lipid composition and particle size affect the structure of specific domains of apo A-I on HDL. PMID- 10801322 TI - Squash trypsin inhibitors from Momordica cochinchinensis exhibit an atypical macrocyclic structure. AB - Three trypsin inhibitors (TIs), from the seeds of the squash Momordica cochinchinensis (MCo), have been isolated and purified using gel filtration, ion exchange chromatography, and reverse-phase HPLC. Their sequences could be determined only after proteolytic cleavages. In the case of MCoTI-I and -II, it was shown that their polypeptide backbones are cyclic, a structure that has never been described in squash TIs. They contain 34 amino acid residues with 3 disulfide bridges and measured molecular masses of 3453.0 and 3480.7, respectively. They are the largest known macrocyclic peptides containing disulfide bridges. Their sequences show strong homology to other squash TIs, suggesting a similar three-dimensional structure and an analogous mechanism of action. A model of MCoTI-II was constructed by analogy to the crystal structure of the complex between bovine trypsin and CMTI-I, indicating that the linker connecting the two termini is flexible and does not impose significant geometrical constraints. This flexibility allows an Asp-Gly peptide bond rearrangement to occur in this region, giving rise to two isoforms of MCoTI-II. Although the importance of cyclization is not clear, it might confer increased stability and resistance to proteolysis. A minor species, MCoTI-III, was also characterized as containing 30 amino acid residues with a molecular mass of 3379.6. This component possesses a linear backbone with a blocked N-terminus. MCoTIs represent interesting candidates for drug design, either by changing their specificity of inhibition or by using their structure as natural scaffolds bearing new binding activities. PMID- 10801323 TI - Conformational modulation of human cytochrome P450 2E1 by ethanol and other substrates: a CO flash photolysis study. AB - The alcohol-inducible cytochrome P450 2E1 is a major human hepatic P450 which metabolizes a broad array of endogenous and exogenous compounds, including ethanol, low-molecular weight toxins, and fatty acids. Several substrates are known to stabilize this P450 and inhibit its cellular degradation. Furthermore, ethanol is a known modulator of P450 2E1 substrate metabolism. We examined the CO binding kinetics of P450 2E1 after laser flash photolysis of the heme-CO bond, to probe the effects of ethanol and other substrates on protein conformation and dynamics. Ethanol had an effect on the two kinetic parameters that describe CO binding: it decreased the rate of CO binding, suggesting a decrease in the protein's conformational flexibility, and increased the photosensitivity, which indicates a local effect in the active site region such as strengthening of the heme-CO bond. Other substrates decreased the CO binding rate to varying degrees. Of particular interest is the effect of arachidonic acid, which abolished photodissociation in the absence of ethanol but had no effect in the presence of ethanol. These results are consistent with a model of P450 2E1 whereby arachidonic acid binds along a long hydrophobic binding pocket and blocks exit of CO from the heme region. PMID- 10801324 TI - Phosphorylation modulates the affinity of light-activated rhodopsin for G protein and arrestin. AB - Reduced effector activity and binding of arrestin are widely accepted consequences of GPCR phosphorylation. However, the effect of receptor multiphosphorylation on G protein activation and arrestin binding parameters has not previously been quantitatively examined. We have found receptor phosphorylation to alter both G protein and arrestin binding constants for light activated rhodopsin in proportion to phosphorylation stoichiometry. Rod disk membranes containing different average receptor phosphorylation stoichiometries were combined with G protein or arrestin, and titrated with a series of brief light flashes. Binding of G(t) or arrestin to activated rhodopsin augmented the 390 nm MII optical absorption signal by stabilizing MII as MII.G or MII.Arr. The concentration of active arrestin or G(t) and the binding constant of each to MII were determined using a nonlinear least-squares (Simplex) reaction model analysis of the titration data. The binding affinity of phosphorylated MII for G(t) decreased while that for arrestin increased with each added phosphate. G(t) binds more tightly to MII at phosphorylation levels less than or equal to two phosphates per rhodopsin; at higher phosphorylation levels, arrestin binding is favored. However, arrestin was found to bind much more slowly than G(t) at all phosphorylation levels, perhaps allowing time for phosphorylation to gradually reduce receptor-G protein interaction before arrestin capping of rhodopsin. Sensitivity of the binding constants to ionic strength suggests that a strong membrane electrostatic component is involved in both the reduction of G(t) binding and the increase of arrestin binding with increasing rhodopsin phosphorylation. PMID- 10801325 TI - Mechanism of oxime reactivation of acetylcholinesterase analyzed by chirality and mutagenesis. AB - Organophosphates inactivate acetylcholinesterase by reacting covalently with the active center serine. We have examined the reactivation of a series of resolved enantiomeric methylphosphonate conjugates of acetylcholinesterase by two oximes, 2-pralidoxime (2-PAM) and 1-(2'-hydroxyiminomethyl-1'-pyridinium)-3-(4'-carbamoyl 1-pyridinium) (HI-6). The S(p) enantiomers of the methylphosphonate esters are far more reactive in forming the conjugate with the enzyme, and we find that rates of oxime reactivation also show an S(p) versus R(p) preference, suggesting that a similar orientation of the phosphonyl oxygen toward the oxyanion hole is required for both efficient inactivation and reactivation. A comparison of reactivation rates of (S(p))- and (R(p))-cycloheptyl, 3,3-dimethylbutyl, and isopropyl methylphosphonyl conjugates shows that steric hindrance by the alkoxy group precludes facile access of the oxime to the tetrahedral phosphorus. To facilitate access, we substituted smaller side chains in the acyl pocket of the active center and find that the Phe295Leu substitution enhances the HI-6-elicited reactivation rates of the S(p) conjugates up to 14-fold, whereas the Phe297Ile substitution preferentially enhances 2-PAM reactivation by as much as 125-fold. The fractional enhancement of reactivation achieved by these mutations of the acyl pocket is greatest for the conjugated phosphonates of the largest steric bulk. By contrast, little enhancement of the reactivation rate is seen with these mutants for the R(p) conjugates, where limitations on oxime access to the phosphonate and suboptimal positioning of the phosphonyl oxygen in the oxyanion hole may both slow reactivation. These findings suggest that impaction of the conjugated organophosphate within the constraints of the active center gorge is a major factor in influencing oxime access and reactivation rates. Moreover, the individual oximes differ in attacking orientation, leading to the presumed pentavalent transition state. Hence, their efficacies as reactivating agents depend on the steric bulk of the intervening groups surrounding the tetrahedral phosphorus. PMID- 10801326 TI - Wheat germ translation initiation factor eIF4B affects eIF4A and eIFiso4F helicase activity by increasing the ATP binding affinity of eIF4A. AB - It has been proposed that, during translational initiation, structures in the 5' untranslated region of mRNA are unwound. eIF4A, a member of the DEAD box family of proteins (those that contain a DEAD amino acid sequence), separately or in conjunction with other eukaryotic initiation factors, utilizes the energy from ATP hydrolysis to unwind these structures. As a step in defining the mechanism of helicase activity in the wheat germ protein synthesis system, we have utilized direct fluorescence measurements, ATPase assays, and helicase assays. The RNA duplex unwinding activity of wheat germ eIF4A is similar to other mammalian systems; however, eIF4F or eIFiso4F is required, probably because of the low binding affinity of wheat germ eIF4A for mRNA. Direct ATP binding measurements showed that eIF4A had a higher binding affinity for ADP than ATP, resulting in a limited hydrolysis and procession along the RNA in the helicase assay. The addition of eIF4B resulted in a change in binding affinity for ATP, increasing it almost 10-fold while the ADP binding affinity was approximately the same. The data presented in this paper suggest that eIF4F or eIFiso4F acts to position the eIF4A and stabilize the interaction with mRNA. ATP produces a conformational change which allows a limited unwinding of the RNA duplex. The binding of eIF4B either prior to or after hydrolysis allows for increased affinity for ATP and for the cycle of conformational changes to proceed, resulting in further unwinding and processive movement along the mRNA. PMID- 10801327 TI - Identification of insulin-induced sites of ribosomal protein S6 phosphorylation in Drosophila melanogaster. AB - Insulin treatment of Drosophila melanogaster Kc 167 cells induces the multiple phosphorylation of a Drosophila ribosomal protein, as judged by its decreased electrophoretic mobility on two-dimensional polyacrylamide gels. The extent to which insulin induces this response is potentiated by cycloheximide and blocked by pretreatment with rapamycin. Isolation and mass spectrometric analysis revealed that the multiply phosphorylated protein was the larger of two Drosophila melanogaster orthologues of mammalian 40S ribosomal protein S6, termed here DS6A. Proteolytic cleavage of DS6A derived from stimulated Kc 167 cells with the endoproteinase Lys-C released a number of peptides, one of which contained all the putative phosphorylation sites. Conversion of phosphoserines to dehydroalanines with Ba(OH)(2) showed that the sites of phosphorylation reside at the carboxy terminus of DS6A. The sites of phosphorylation were identified by Edman degradation after conversion of the phosphoserine residues to S ethylcysteine as Ser(233), Ser(235), Ser(239), Ser(242), and Ser(245). Finally, phosphopeptide mapping of individual phosphoderivatives, isolated from two dimensional polyacrylamide gels, indicated that DS6A phosphorylation, in analogy to mammalian S6 phosphorylation, appears to proceed in an ordered fashion. The importance of these observations in cell growth and development is discussed. PMID- 10801328 TI - Mutational analysis of the epimerization domain in the initiation module PheATE of gramicidin S synthetase. AB - The epimerase (E) domain of the three-domain (ATE) initiation module of Bacillus brevis gramicidin S synthetase equilibrates the Calpha configuration of the phenylalanyl moiety presented as Phe-S-4'-phosphopantetheine-modified (Ppant) acyl enzyme. Mutants at 22 residues of this E domain that are conserved across the approximately 450 residue E domains of nonribosomal peptide synthetases were constructed, and the PheATE derivatives expressed in Escherichia coli as C terminal His tag fusions and then purified and assayed for three activities: (1) the L-Phe Calpha-[(3)H] exchange to solvent, (2) the rate of approach to D-Phe/L Phe-S-Ppant acyl enzyme equilibrium from either L- or D-Phe, and (3) the rate of Phe-Pro dipeptidyl-S-Ppant enzyme formation with the downstream ProCAT module. We found that for wild-type PheATE epimerization is much faster than subsequent condensation, leading to a 1.9:1 ratio of D-Phe-S-Ppant/L-Phe-S-Ppant acyl enzyme. Only D-Phe is then transferred to yield D-Phe-L-Pro-S-Ppant ProCAT acyl enzyme. Among the mutants generated, three PheATE constructs, H753A, D757S, and Y976A, showed no detectable Calpha-(3)H washout, while E892A and R896A were among a larger set partially impaired. All these mutants were dramatically impaired in approach to D-Phe/L-Phe-S-Ppant equilibrium from either D- or L-Phe, while another construct, D767S, was asymmetrically impaired only for D-to-L-Phe direction. In the D-Phe-L-Pro dipeptidyl-S-Ppant condensation assay, the H753A and E892A forms of PheATE were only slightly active from L-Phe but unimpaired from D-Phe; N975A epimerizes faster than Y976A from L-Phe. When the chirality of the Phe-Pro-diketopiperazine released product was analyzed the D,L/L,L ratio from wild-type PheATE and ProCAT was 98:2. From E892A and N975A it was comparably 95:5 and 92:8, but H753A and Y976A yielded 56% of the L,L-product, reflecting a gain of function to transfer L-Phe. The 98:2 preference of wild-type PheATE for D-Phe transfer reflects the kinetically controlled stereopreference of the condensation (C) domain of ProCAT for the D-Phe-S-Ppant donor substrate. It may be that other NRPS C domains immediately downstream of E domains will likewise be D-selective. PMID- 10801329 TI - Ultraviolet damage endonuclease (Uve1p): a structure and strand-specific DNA endonuclease. AB - Schizosaccharomyces pombe ultraviolet damage endonuclease (UVDE or Uve1p) performs the initial step in an alternative excision repair pathway for UV induced DNA damage. This DNA repair pathway was originally thought to be specific for UV damage. However, the broad substrate specificity of Uve1p suggests a more general role for this enzyme. Uve1p recognizes UV-induced bipyrimidine photoadducts and other non-UV-induced DNA adducts. Biochemical and genetic analysis also suggests that Uve1p may be involved in orchestrating mismatch repair in vivo. This study demonstrates that Uve1p recognizes and cleaves heteroduplex DNA with small unpaired loops but does not recognize loops six to eight nucleotides in length. In addition, the enzyme does not recognize DNA with palindromic insertions that could form base-paired hairpin structures. The cleavage efficiency of Uve1p depends on the distance of a mismatch from the DNA terminus, suggesting that the 3' terminus may contribute to the strand discrimination signal for Uve1p. These biochemical activities are discussed in the context of the role of Uve1p in DNA repair. PMID- 10801330 TI - Tyrosine dephosphorylation, but not phosphorylation, of p130Cas is dependent on integrin alpha IIb beta 3-mediated aggregation in platelets: implication of p130Cas involvement in pathways unrelated to cytoskeletal reorganization. AB - The newly described adapter molecule p130 Crk-associated substrate (Cas) has been reported to contribute to cytoskeletal organization through assembly of actin filaments and to be pivotal in embryonic development and in oncogene-mediated transformation. We characterized the regulation of Cas tyrosine phosphorylation in highly differentiated, anucleate platelets. Phospholipase C-activating receptor agonists, including collagen, thrombin receptor-activating peptide (TRAP), and U46619 (a thromboxane A2 analogue), and A23187 (a Ca2+ ionophore) induced rapid Cas tyrosine phosphorylation in platelets. 12-O Tetradecanoylphorbol 13-acetate and 1-oleoyl-2-acetyl-sn-glycerol, protein kinase C (PKC) activators, also induced Cas tyrosine phosphorylation, albeit sluggishly. Cas tyrosine phosphorylation induced by collagen or TRAP was transient in aggregating platelets; Cas became dephosphorylated in a manner dependent on integrin alpha IIb beta 3-mediated aggregation. While BAPTA-AM (an intracellular Ca2+ chelator) inhibited Cas phosphorylation induced by collagen or TRAP, Ro31 8220 (a PKC inhibitor) rather prolonged it. Under the conditions, this PKC inhibitor suppressed platelet aggregation but not intracellular Ca2+ mobilization. In contrast to Cas involvement in focal adhesions in other cells, platelet Cas phosphorylation preceded the activation of focal adhesion kinase (FAK), and blockage of alpha IIb beta 3-mediated platelet aggregation with a GRGDS peptide resulted in prolongation of stimulation-dependent Cas tyrosine phosphorylation but in suppression of FAK tyrosine phosphorylation. Furthermore, TRAP-induced Cas phosphorylation was insensitive to cytochalasin D, an actin polymerization inhibitor. The failure of FAK to associate with Cas in immunoprecipitation studies also suggests that Cas tyrosine phosphorylation is independent of FAK activation. Of the signaling molecules investigated in this study, Src seemed to associate with Cas. Finally, Cas existed mainly in cytosol and membrane cytoskeleton fractions in the resting state, and remained unchanged during platelet aggregation, when FAK translocated to the cytoskeletal fraction. Our findings on platelet Cas suggest that (i) rapid Cas tyrosine phosphorylation occurs following phosphoinositide turnover by receptor-mediated agonists and may be mediated by intracellular Ca2+ mobilization; (ii) PKC activation, by itself, may elicit sluggish Cas phosphorylation; (iii) Cas tyrosine dephosphorylation, but not phosphorylation, is dependent on integrin alpha IIb beta 3-mediated aggregation; and (iv) Cas is not involved in cytoskeletal reorganization. Anucleate platelets seem to provide a unique model system to fully elucidate the functional role(s) of Cas. PMID- 10801331 TI - A plausible mechanism for gene correction by chimeric oligonucleotides. AB - Self-complementary chimeric oligonucleotides that consist of DNA and 2'-O-methyl RNA nucleotides arranged in a double-hairpin configuration can elicit a point mutation when targeted to a gene sequence. We have used a series of structurally diverse chimeric oligonucleotides to correct a mutant neomycin phosphotransferase gene in a human cell-free extract. Analysis of structure-activity relationships demonstrates that the DNA strand of the chimeric oligonucleotide acts as a template for high-fidelity gene correction when one of its bases is mismatched to the targeted gene. By contrast, the chimeric strand of the oligonucleotide does not function as a template for gene repair. Instead, it appears to augment the frequency of gene correction by facilitating complex formation with the target. In the presence of RecA protein, each strand of a chimeric oligonucleotide can hybridize with double-stranded DNA to form a complement-stabilized D-loop. This reaction, which may take place by reciprocal four-strand exchange, is not observed with oligonucleotides that lack 2'-O-methyl RNA segments. Preliminary sequencing data suggest that complement-stabilized D-loops may be weakly mutagenic. If so, a low level of random mutagenesis in the vicinity of the chimera binding site may accompany gene repair. PMID- 10801332 TI - Iminium ion chemistry of mitosene DNA alkylating agents. Enriched 13C NMR and isolation studies. AB - Described herein is a study of the reductive alkylation chemistry of mitosene antitumor agents. We employed a 13C-enriched electrophilic center to probe the fate of the iminium ion resulting from reductive activation. The 13C-labeled center permitted the identification of complex products resulting from alkylation reactions. In the case of DNA reductive alkylation, the type and number of alkylation sites were readily assessed by 13C NMR. Although there has been much excellent work done in the area of mitosene chemistry and biochemistry, the present study provides a number of new findings: (1) The major fate of the iminium ion is head-to-tail polymerization, even in dilute solutions. (2) Dithionite reductive activation results in the formation of mitosene sulfite esters as well as the previously observed sulfonate adducts. (3) The mitosene iminium ion alkylates the adenosine 6-amino group as well as the guanosine 2 amino group. The identification of the latter adduct was greatly facilitated by the 13C-label at the electrophilic center. (4) The mitosene iminium ion alkylates DNA at both nitrogen and oxygen centers without any apparent base selectivity. The complexity of mitosene reductive alkylation of DNA will require continued adduct isolation studies. PMID- 10801333 TI - Identification of mutations in rat CD59 that increase the complement regulatory activity. AB - Formation of the membrane attack complex (MAC) of complement on host cells is inhibited by the glycosylphosphatidylinositol- (GPI-) anchored glycoprotein CD59. Published data on the active site of human CD59 are confusing. To clarify these data, we set out to elucidate the active site of a nonprimate CD59 molecule by site-directed mutagenesis. We also undertook to investigate a region of potential species selectivity, and to this end rat CD59 was chosen for all mutations. Our investigations confirmed the proposal that the active site of CD59 is the major hydrophobic groove, with mutations Y36A, W40A, and L54A ablating complement inhibitory function of CD59. Other mutations reducing the function of rat CD59 were I56E, D24A, and D24R. Importantly, mutations at one residue increased the function of rat CD59. The K48E mutation significantly increased function against human rat or rabbit serum, whereas the K48A mutation increased function against human serum alone. A similar mutation in human CD59 (N48E) had no effect on activity against human or rat serum but completely abolished all activity against rabbit serum. These findings suggest that the alpha-helix of human CD59, adjacent to the hydrophobic groove, influences the interaction between human CD59 and rabbit C8, C9, or both. PMID- 10801334 TI - Phosphatidylinositol 4,5-bisphosphate domain inducers promote phospholipid transverse redistribution in biological membranes. AB - Transmembrane phospholipid redistribution (scrambling), leading to exposure of phosphatidylserine on the cell surface, plays a physiological role to induce platelet procoagulant activity and clearance of injured or apoptotic cells. Scrambling is generally attributed to an increase in intracellular Ca(2+) and would be mediated by a protein (scramblase), whose activity could be modulated by cofactors. We reported previously that phosphatidylinositol 4,5-bisphosphate (PIP(2)) is a positive regulator of Ca(2+)-induced scrambling. We show here, using inside-out vesicles from erythrocyte membranes, that a pleckstrin homology (PH) domain, which interacts with high affinity with PIP(2), inhibited Ca(2+) induced scrambling, confirming the role of PIP(2). As Ca(2+) is known to interact with PIP(2) and to promote the formation of lateral domains of acidic phospholipids in membranes, we investigated whether PIP(2) domain formation could be involved in scrambling. Spermine, polylysine, and MARCKS (151-175) peptide caused scrambling in parallel to their reported ability to form domains of acidic phospholipids, including PIP(2). Similarly, neomycine, another PIP(2)-interacting polycation, induced scrambling. A PIP(2) antibody was also found to induce scrambling, presumably by a similar mechanism, since phospholipid antibodies are known to promote phospholipid capping. In conclusion, Ca(2+) is not the sole inducer of scrambling, and formation of PIP(2) domains could play a critical role in this process. PMID- 10801335 TI - Uncoupling proteins 1 and 3 are regulated differently. AB - Using a heterologous yeast expression system, we have previously found a marked discordance between the effects of uncoupling protein (UCP) 1 and UCP3L on basal O(2) consumption in whole yeast versus isolated mitochondria. In whole yeast, UCP3L produces a greater stimulation of basal O(2) consumption, while in isolated mitochondria, UCP1 produces a much greater effect. As shown previously and in this report, UCP3L, in contrast to UCP1, is not inhibited by purine nucleotides. In the present study, we addressed two hypothetical mechanisms that could account for the observed discordance: (i) in whole yeast, purine nucleotides inhibit UCP1 but not UCP3L and (ii) preparations of isolated mitochondria lack an activator of UCP3L that is normally present in vivo. By use of a mutant of UCP1 that lacks purine nucleotide inhibition, it is demonstrated that cytosolic concentrations of purine nucleotides present in yeast effectively inhibit UCP1 activity. This suggests that the lower activity of UCP1 compared to UCP3L in whole yeast is due to purine nucleotide inhibition of UCP1 but not UCP3L. As potential activators of UCP3L we tested free fatty acids in whole yeast and isolated mitochondria. While UCP1 was strongly activated by free fatty acids, no stimulatory effect on UCP3L was observed. In summary, this study indicates that UCP1 and UCP3L differ in their regulation by purine nucleotides and free fatty acids. This different regulation may be related to different physiological functions of the two proteins. PMID- 10801336 TI - Visualization of highly ordered striated domains induced by transmembrane peptides in supported phosphatidylcholine bilayers. AB - We used atomic force microscopy (AFM) to study the lateral organization of transmembrane TmAW(2)(LA)(n)W(2)Etn peptides (WALP peptides) incorporated in phospholipid bilayers. These well-studied model peptides consist of a hydrophobic alanine-leucine stretch of variable length, flanked on each side by two tryptophans. They were incorporated in saturated phosphatidylcholine (PC) vesicles, which were deposited on a solid substrate via the vesicle fusion method, yielding hydrated gel-state supported bilayers. At low concentrations (1 mol %) WALP peptides induced primarily line-type depressions in the bilayer. In addition, striated lateral domains were observed, which increased in amount and size (from 25 nm up to 10 microm) upon increasing peptide concentration. At high peptide concentration (10 mol %), the bilayer consisted mainly of striated domains. The striated domains consist of line-type depressions and elevations with a repeat distance of 8 nm, which form an extremely ordered, predominantly hexagonal pattern. Overall, this pattern was independent of the length of the peptides (19-27 amino acids) and the length of the lipid acyl chains (16-18 carbon atoms). The striated domains could be pushed down reversibly by the AFM tip and are thermodynamically stable. This is the first direct visualization of alpha-helical transmembrane peptide-lipid domains in a bilayer. We propose that these striated domains consist of arrays of WALP peptides and fluidlike PC molecules, which appear as low lines. The presence of the peptides perturbs the bilayer organization, resulting in a decrease in the tilt of the lipids between the peptide arrays. These lipids therefore appear as high lines. PMID- 10801337 TI - Influence of monovalent cations on rat alpha- and beta-parvalbumin stabilities. AB - The mammalian genome encodes both alpha- and beta-parvalbumin isoforms. The rat beta-parvalbumin (aka "oncomodulin") is more stable than the alpha isoform at physiological pH and ionic strength, despite its substantially higher charge density and truncated C-terminal helix [Henzl, M. T., and Graham, J. S. (1999) FEBS Lett. 442, 241-245]. Reasoning that solvent interactions could contribute to this unexpected finding, we have examined the stabilities of the Ca(2+)-free alpha- and beta-parvalbumins as a function of Na(+) and K(+) concentration. Differential scanning calorimetry data suggest that, at physiological pH and ionic strength, the beta isoform binds roughly 2 equiv of Na(+) or a single equivalent of K(+) with moderate affinity. Under comparable conditions, the alpha isoform apparently binds just 1 equiv of Na(+) and essentially no K(+). Isothermal titration calorimetry experiments suggest that the bound monovalent ions occupy the EF-hand motifs. In 0.15 M K(+), at pH 7.4, the stability of the apo-beta-parvalbumin exceeds that of the alpha isoform by approximately 2.6 kcal/mol at 37 degrees C and by approximately 3.0 kcal/mol at 25 degrees C. The latter value represents a substantial fraction of the difference in Ca(2+) binding free energies measured in vitro for the two proteins. Significantly, however, these results do not completely explain the paradoxical stability of the beta isoform, which maintains its higher melting temperature under all conditions examined. PMID- 10801338 TI - Modulation of the activities of catalase-peroxidase HPI of Escherichia coli by site-directed mutagenesis. AB - Catalase-peroxidases have a predominant catalatic activity but differ from monofunctional catalases in exhibiting a substantial peroxidatic reaction which has been implicated in the activation of the antitubercular drug isoniazid in Mycobacterium tuberculosis. Hydroperoxidase I of Escherichia coli encoded by katG is a catalase-peroxidase, and residues in its putative active site have been the target of a site directed-mutagenesis study. Variants of residues R102 and H106, on the distal side of the heme, and H267, the proximal side ligand, were constructed, all of which substantially reduced the catalatic activity and, to a lesser extent, the peroxidatic activity. In addition, the heme content of the variants was reduced relative to the wild-type enzyme. The relative ease of heme loss from HPI and a mixture of tetrameric enzymes with 2, 3, and 4 hemes was revealed by mass spectrometry analysis. Conversion of W105 to either an aromatic (F) or aliphatic (I) residue caused a 4-5-fold increase in peroxidatic activity, coupled with a >99% inhibition of catalatic activity. The peroxidatic-to catalatic ratio of the W105F variant was increased 2800-fold such that compound I could be identified by both electronic and EPR spectroscopy as being similar to the porphyrin cation radical formed in other catalases and peroxidases. Compound I, when generated by a single addition of H(2)O(2), decayed back to the native or resting state within 1 min. When H(2)O(2) was generated enzymatically in situ at low levels, active compound I was evident for up to 2 h. However, such prolonged treatment resulted in conversion of compound I to a reversibly inactivated and, eventually, to an irreversibly inactivated species, both of which were spectrally similar to compound I. PMID- 10801339 TI - Evidence for the involvement of histidine A(12) in the aggregation and precipitation of human relaxin induced by metal-catalyzed oxidation. AB - The metal-catalyzed oxidation (ascorbate/cupric chloride/oxygen) of recombinant human relaxin (rhRlx, type II) was shown by Li et al. [Li, S., Nguyen, T. H., Schoneich, C., and Borchardt, R. T. (1995) Biochemistry 34, 5762-5772] to result in the chemical modification of His A(12), Met B(4), and Met B(25). Considering the fact that His A(12) exists in an extended loop that joins two alpha-helices in this protein, we hypothesized that oxidation of this specific amino acid leads to alterations in the secondary and tertiary structures of the protein, resulting in the pH-dependent aggregation/precipitation phenomena observed in our earlier studies (i.e., at pH >6.0 most of the degradants of rhRlx are insoluble). Evidence obtained in the current study that supports this hypothesis includes the following: (i) oxidation of rhRlx with hydrogen peroxide (H(2)O(2)), which leads only to modification of Met B(4) and Met B(25), does not result in the pH dependent aggregation/precipitation of the protein; and (ii) metal-catalyzed oxidation of porcine relaxin (pRlx), which does not contain His at position A(12), leads to chemical degradation of the protein [e.g., Met A(2) is oxidized] but produces only slight pH-dependent aggregation/precipitation of the protein. In addition, experimental evidence is provided to show that the physical instability of rhRlx observed at pH >6.0 does not appear to be related to the pH dependent solubility of a common protein degradant. Instead, it appears that several oxidation products of His A(12) are produced in a pH-dependent manner and that these oxidation products produce different effects on the physical stability of the protein. Evidence in support of this conclusion includes the observation that the soluble degradants of rhRlx showed reduced levels of His, reduced levels of the T(2)-T(7) tryptic fragment that contained His A(12), and the presence of 2 oxo-His. Similarly, the precipitated degradants of rhRlx showed reduced levels of His but no 2-oxo-His. In addition, the soluble degradants, which contain 2-oxo His, appear to exist as monomers having an average molecular weight similar to that of rhRlx. These results suggest that the metal-catalyzed oxidation of His A(12) leads to other, as yet unidentified oxidation products of His A(12) that affect the secondary/tertiary structure of the protein more significantly than does 2-oxo-His and ultimately lead to the physical instability of the protein observed at higher pH values. PMID- 10801340 TI - Influence of sequence-dependent cytosine protonation and methylation on DNA triplex stability. AB - To investigate cytosine protonation and its influence on the sequence-dependent thermal stability of DNA triplexes in detail, we have employed homo- and heteronuclear NMR experiments on specifically (15)N-labeled oligodeoxynucleotides that were designed to fold into intramolecular triple helices of the pyrimidine motif under appropriate conditions. These experiments reveal that cytosines in central positions of the triplex are significantly protonated even at neutral pH. However, semiprotonation points for individual cytosine bases as determined from pH-dependent measurements show considerable differences depending on their position. Thus, protonation is disfavored for adjacent cytosines or for cytosines at the triplex termini, resulting in a smaller contribution to the overall free energy of the triple helical system. In contrast, protonation of the base upon substitution of 5-methylcytosine for cytosine in the triplex third strand is only affected to a minor extent, and triplex stabilization by the methyl substituent is shown to primarily arise from stacking energies and/or hydrophobic effects. PMID- 10801341 TI - Binding of the PH and polybasic C-terminal domains of ARNO to phosphoinositides and to acidic lipids. AB - The activity on ARF of the guanine nucleotide exchange factor ARNO depends on its membrane recruitment, induced by binding of its PH domain to phosphoinositides. A polycationic C-terminal extension to the PH domain might also contribute to its specific binding to phosphatidylinositol 4,5-bisphosphate [(4,5)PIP2] and to phosphatidylinositol 3,4,5-trisphosphate [(3,4,5)PIP3], and to ionic binding to other acidic lipids. We have analyzed in vitro the relative contributions to phospholipid binding of the PH domain and C-terminal extension by cosedimentation of "PH+C domain" and "nominal PH domain" protein constructs including or not including the polycationic C-terminus, with sucrose-loaded unilamellar vesicles made of equal proportions of the neutral lipids phosphatidylcholine and phosphatidylethanolamine, and supplemented or not with 30% acidic phosphatidylserine (PS) and 2% of various phosphoinositides. Binding was measured as a function of the vesicle concentration and of the medium ionic strength. Both proteins bound with higher affinity to (3,4,5)PIP3 than to (4,5)PIP2, the selectivity for (3,4,5)PIP3 being highest for the nominal PH domain. We observed also a clear selectivity of (3,4,5)PIP3 over (4,5)PIP2 for stimulating the activity of ARNO on ARF with vesicles containing 10% PS and 1% PIP2 or PIP3. Our data suggest that the PH domain provides the specific phosphoinositide binding site and some unspecific ionic interaction with acidic PS, whereas the polybasic C domain contributes to binding mainly by unspecific ionic interactions vith PS. Phosphorylation by protein kinase C of a serine in the C domain reduces the ionic affinity of the PH+C domain for PS, but does not affect the phosphoinositide specificity. PMID- 10801342 TI - Primary 13C and beta-secondary 2H KIEs for trans-sialidase. A snapshot of nucleophilic participation during catalysis. AB - Trypanosoma cruzi trans-sialidase catalyzes a novel reaction that involves the transfer of sialic acid between host and parasite glycoconjugates. In this paper, we report kinetic isotope effect studies on recombinant trans-sialidase. beta Dideuterium and primary 13C isotope effects were measured for a good substrate, sialyl-lactose, and a slow substrate, sialyl-galactose, in both acid-catalyzed solvolysis and enzymatic transfer reactions. The beta-dideuterium isotope effect for sialyl-lactose in the acid hydrolysis reaction was 1.113 +/- 0.012. The primary 13C isotope effects for hydrolysis of sialyl-lactose and sialyl-galactose were 1. 016 +/- 0.011 and 1.015 +/- 0.008, respectively. In the enzymatic transfer reactions, the beta-dideuterium and primary 13C effects for sialyl galactose were 1.060 +/- 0.008 and 1.032 +/- 0.008, respectively. The isotope effects for hydrolysis describe a dissociative SN1-like mechanism, and these data are contrasted by the data for the enzyme-catalyzed reaction. The enzymatic deuterium isotope effects are lower by a factor of 2, but the primary carbon isotope effects are higher by a factor of 2. This pattern describes a mechanism involving nucleophilic participation in the rate-determining transition state. PMID- 10801343 TI - Identification of initiation sites for T4 lysozyme folding using CD and NMR spectroscopy of peptide fragments. AB - Using CD and 2D (1)H NMR spectroscopy, we have identified potential initiation sites for the folding of T4 lysozyme by examining the conformational preferences of peptide fragments corresponding to regions of secondary structure. CD spectropolarimetry showed most peptides were unstructured in water, but adopted partial helical conformations in TFE and SDS solution. This was also consistent with the (1)H NMR data which showed that the peptides were predominantly disordered in water, although in some cases, nascent or small populations of partially folded conformations could be detected. NOE patterns, coupling constants, and deviations from random coil Halpha chemical shift values complemented the CD data and confirmed that many of the peptides were helical in TFE and SDS micelles. In particular, the peptide corresponding to helix E in the native enzyme formed a well-defined helix in both TFE and SDS, indicating that helix E potentially forms an initiation site for T4 lysozyme folding. The data for the other peptides indicated that helices D, F, G, and H are dependent on tertiary interactions for their folding and/or stability. Overall, the results from this study, and those of our earlier studies, are in agreement with modeling and HD-deuterium exchange experiments, and support an hierarchical model of folding for T4 lysozyme. PMID- 10801344 TI - Mimicry of human IgE epitopes by anti-idiotypic antibodies. AB - According to Jerne's network hypothesis, the binding site of an anti-idiotypic antibody also represents the internal image of an epitope present on a foreign, or even a self antigen. In recent years, antigen mimicry has been defined at the molecular level for some xeno-antigens. However, until now there has been no demonstration of structural mimicry between a human anti-idiotypic antibody and a self structure. To address this question, we used human IgE as the self structure and a well-defined anti-human IgE mAb (BSW17). We describe the isolation of two anti- idiotypic antibodies specific for the anti-IgE antibody BSW17 from a non immune human Fab phage display library. Interestingly, these two anti-idiotypic antibodies mimic the same molecular surface region as a previously described IgE peptide mimotope isolated by panning on BSW17, but they cover a much larger epitope on the IgE molecule. Accordingly, immunisation of rabbits with the two anti-idiotypic antibodies induced high-affinity antibodies with the same characteristics as BSW17. Thus, our data demonstrate that it is possible to isolate anti-idiotypic antibodies derived from the human genome without the need for hyperimmunization, and confirm Jerne's hypothesis that both foreign antigens and self structures can be mimicked by our own immunoglobulins. PMID- 10801345 TI - The bacterial DNA-binding protein H-NS represses ribosomal RNA transcription by trapping RNA polymerase in the initiation complex. AB - The interaction of the bacterial regulatory protein H-NS with RNA polymerase and the ribosomal RNA P1 promoter was analyzed to better understand the mechanism of H-NS-dependent transcriptional repression. We could show that initial binding of RNA polymerase to the promoter was not inhibited by the simultaneous interaction of H-NS, although H-NS binding sites extend into the core promoter region. Binding of sigma(70)-saturated RNA polymerase and H-NS to the promoter DNA occurs cooperatively and results in a stable complex of slower gel electrophoretic mobility as compared to complexes formed with the single proteins. The presence of the upstream curved H-NS binding site contributes strongly to the cooperative RNA polymerase-promoter interaction. By KMnO(4) modification of single-stranded template nucleotides we could show that open complex formation at the rrnB P1 promoter was not inhibited by H-NS binding. An increased KMnO(4) reactivity of several positions within the open complex rather supports the view that open complex formation is stimulated in presence of H-NS. Moreover, subtle changes in the modification pattern indicate that the open complex formed in the presence of H-NS are structurally distinct from the H-NS-free complex. In vitro transcriptional analysis of the abortive and productive yields revealed that the formation of transcription products longer than three nucleotides is dramatically reduced in the presence of H-NS, while the amount of shorter abortive products remained unaffected. Together the results demonstrate that H-NS inhibits transcription at the rrnB P1 promoter not by interfering with initial RNA polymerase binding but by blocking chain elongation steps subsequent to the first (two) phosphodiester bond formations. The mechanism of H-NS dependent repression at rRNA promoters can thus be explained as a trap which inhibits substrate NTP incorporation beyond template position +3 into the initial transcribing complex. PMID- 10801346 TI - Geometry of site alignment during int family recombination: antiparallel synapsis by the Flp recombinase. AB - The Flp site-specific recombinase functions in the copy number amplification of the yeast 2 microm plasmid. The recombination reaction is catalyzed by four monomers of Flp bound to two separate, but identical, recombination sites (FRT sites) and occurs in two sequential pairs of strand exchanges. The relative orientation of the two recombination sites during synapsis was examined. Topoisomerase relaxation and nick ligation were used to detect topological nodes introduced by the synapse prior to the chemical steps of recombination. A single negative supercoil was found to be trapped by Flp in substrates with inverted FRT sites whereas no trapped supercoils were observed with direct repeats. The topology of products resulting from Flp-mediated recombination adjacent to a well characterised synapse, that of Tn3 resolvase/res, was analyzed. The deletion and inversion reactions yielded the four noded catenane and the three noded knot, respectively, as the simplest and the most abundant products. The linking number change introduced by the Flp-mediated inversion reaction was determined to be +/ 2. The most parsimonious explanation of these results is that Flp aligns its recombination sites with antiparallel geometry. The majority of synapses appear to occur without entrapment of additional random plectonemic DNA supercoils between the sites and no additional crossings are introduced as a result of the chemical steps of recombination. PMID- 10801347 TI - Binding of the Epstein-Barr virus major envelope glycoprotein gp350 results in the upregulation of the TNF-alpha gene expression in monocytic cells via NF kappaB involving PKC, PI3-K and tyrosine kinases. AB - Epstein-Barr virus (EBV) is a human herpesvirus that interacts with various immunocompetent cells that carry the EBV receptor (CD21/CR2). EBV binds to CR2 through its major envelope glycoprotein 350 (gp350). Previously we had demonstrated that EBV and other human herpesviruses are capable of modulating cytokine synthesis through the deregulated expression of cytokine genes interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF alpha), and interleukin-2 (IL-2). Here we show that, in contrast to infectious EBV, purified recombinant gp350 upregulates TNF-alpha gene expression in human monocyte/macrophages (M/M) as well as in a monocytoid cell line, U937. Our results also demonstrate that this increased expression is due to both enhanced transcription and stability of TNF-alpha mRNA in gp350-treated cells. The specificity of this effect is evidenced by the fact that pre-incubation of cells with anti-CR2 monoclonal antibody OKB7, which blocks binding of gp350 to CR2, inhibits the above mentioned effects of gp350. Furthermore, we demonstrate that activation of TNF-alpha by gp350 is mediated by NF-kappaB through signal transduction pathways involving PKC, PI3-K and tyrosine kinases. To our knowledge this is the first report describing the modulation of TNF-alpha gene expression by the EBV-gp350 molecule following its interaction with the viral receptor CR2 on cells of the monocytic lineage. PMID- 10801348 TI - tRNA leucine identity and recognition sets. AB - Transfer RNAs (tRNAs) are grouped into two classes based on the structure of their variable loop. In Escherichia coli, tRNAs from three isoaccepting groups are classified as type II. Leucine tRNAs comprise one such group. We used both in vivo and in vitro approaches to determine the nucleotides that are required for tRNA(Leu) function. In addition, to investigate the role of the tRNA fold, we compared the in vivo and in vitro characteristics of type I tRNA(Leu) variants with their type II counterparts.A minimum of six conserved tRNA(Leu) nucleotides were required to change the amino acid identity and recognition of a type II tRNA(Ser) amber suppressor from a serine to a leucine residue. Five of these nucleotides affect tRNA tertiary structure; the G15-C48 tertiary "Levitt base pair" in tRNA(Ser) was changed to A15-U48; the number of nucleotides in the alpha and beta regions of the D-loop was changed to achieve the positioning of G18 and G19 that is found in all tRNA(Leu); a base was inserted at position 47n between the base-paired extra stem and the T-stem; in addition the G73 "discriminator" base of tRNA(Ser) was changed to A73. This minimally altered tRNA(Ser) exclusively inserted leucine residues and was an excellent in vitro substrate for LeuRS. In a parallel experiment, nucleotide substitutions were made in a glutamine-inserting type I tRNA (RNA(SerDelta); an amber suppressor in which the tRNA(Ser) type II extra-stem-loop is replaced by a consensus type I loop). This "type I" swap experiment was successful both in vivo and in vitro but required more nucleotide substitutions than did the type II swap. The type I and II swaps revealed differences in the contributions of the tRNA(Leu) acceptor stem base pairs to tRNA(Leu) function: in the type I, but not the type II fold, leucine specificity was contingent on the presence of the tRNA(Leu) acceptor stem sequence. The type I and II tRNAs used in this study differed only in the sequence and structure of the variable loop. By altering this loop, and thereby possibly introducing subtle changes into the overall tRNA fold, it became possible to detect otherwise cryptic contributions of the acceptor stem sequence to recognition by LeuRS. Possible reasons for this effect are discussed. PMID- 10801349 TI - The phenotype of mutations of the base-pair C2658.G2663 that closes the tetraloop in the sarcin/ricin domain of Escherichia coli 23 S ribosomal RNA. AB - The sarcin/ricin domain (SRD) in Escherichia coli 23 S rRNA is a part of the site for the association of elongation factors with ribosomes and for that reason is critical for the binding of aminoacyl-tRNA and for translocation during the reiterative elongation reactions of protein synthesis. The SRD has a GAGA tetraloop that is shut off by a Watson-Crick C2658 x G2663 pair. The contribution of this pair to the function of the ribosome has been evaluated by constructing mutations in the nucleotides and determining their phenotype. Constitutive expression of a plasmid-encoded rrnB operon with a G2663C transversion mutation that disrupts the Watson-Crick pair was lethal. Double transversion mutations, C2658G x G2663C and C2658A x G2663U, that reverse the polarity of the pyrimidine and the purine but restore the potential to form a canonical pair, were also lethal. Induction of transcription of 23 S rRNA with the same mutations, but encoded in a plasmid with a lambdaP(L) promoter and expressed at a lower level, retarded growth. The sedimentation profiles of ribosomes with transversion mutations in C2658 and/or G2663 are altered; the ratio of 50 S subunits to 30 S particles is changed and polysomes are reduced. Ribosomes with a G2663C, a C2658G x G2663C, or a C2658A x G2663U mutation in 23 S rRNA were not active in protein synthesis, indeed, they appeared to inhibit the activity of ribosomes with wild type 23 S rRNA. Transversion mutations in the analogs of C2658 and G2663 decreased binding of EF-G to SRD oligoribonucleotides; the same mutations in 23 S rRNA decreased binding of the factor to intact ribosomes. The most severe phenotype, in growth, in protein synthesis, and in the binding of EF-G, was associated with a C2658G x G2663C mutation; it is surprising that this was more severe than an analogous C2658A x G2663U mutation. A double transition mutation, C2658U x G2663A, which is not known to have occurred in nature, had no effect on the growth of cells or on the function of ribosomes. The lethal phenotype of transversion mutations in C2658 and G2663 appears to derive from a loss of the capacity of ribosomes to bind EF-G and by indirection the EF-Tu ternary complex. PMID- 10801350 TI - Topology of the components of the DNA packaging machinery in the phage phi29 prohead. AB - Chromosome condensation inside dsDNA viral particles is a complex process requiring the coordinated action of several viral components. The similarity of the process in different viral systems has led to the suggestion that there is a common underlying mechanism for DNA packaging, in which the portal vertex or connector plays a key role. We have studied the topology of the packaging machinery using a number of antibodies directed against different domains of the connector. The charged amino-terminal, the carboxyl-terminal, and the RNA binding domain are accessible areas in the connector assembled into the prohead, while the domains corresponding to the 12 large appendages of the connector are buried inside the prohead. Furthermore, while the antibodies against the carboxyl and amino-terminal do not affect the packaging reaction, incubation of proheads with antibodies against the RNA binding domain abolishes the packaging activity. The comparison of the three-dimensional reconstructions of bacteriophage phi29 proheads with proheads devoid of their specific pRNA by RNase treatment shows that this treatment removes structural elements of the distal vertex of the portal structure, suggesting that the pRNA required for packaging is located at the open gate of the channel in the narrow side of the connector. PMID- 10801351 TI - The intermediate filament protein consensus motif of helix 2B: its atomic structure and contribution to assembly. AB - Nearly all intermediate filament proteins exhibit a highly conserved amino acid motif (YRKLLEGEE) at the C-terminal end of their central alpha-helical rod domain. We have analyzed its contribution to the various stages of assembly by using truncated forms of Xenopus vimentin and mouse desmin, VimIAT and DesIAT, which terminate exactly before this motif, by comparing them with the wild-type and tailless proteins. It is surprising that in buffers of low ionic strength and high pH where the full-length proteins form tetramers, both VimIAT and DesIAT associated into various high molecular weight complexes. After initiation of assembly, both VimIAT and DesIAT aggregated into unit-length-type filaments, which rapidly longitudinally annealed to yield filaments of around 20 nm in diameter. Mass measurements by scanning transmission electron microscopy revealed that both VimIAT and DesIAT filaments contained considerably more subunits per cross-section than standard intermediate filaments. This indicated that the YRKLLEGEE-motif is crucial for the formation of authentic tetrameric complexes and also for the control of filament width, rather than elongation, during assembly. To determine the structure of the YRKLLEGEE domain, we grew crystals of peptides containing the last 28 amino acid residues of coil 2B, chimerically fused at its amino-terminal end to the 31 amino acid-long leucine zipper domain of the yeast transcription factor GCN4 to facilitate appropriate coiled-coil formation. The atomic structure shows that starting from Tyr400 the two helices gradually separate and that the coiled coil terminates with residue Glu405 while the downstream residues fold away from the coiled-coil axis. PMID- 10801352 TI - In-situ atomic force microscopy study of beta-amyloid fibrillization. AB - We report the use of atomic force microscopy to observe the initial stages of beta-amyloid fibrillization in situ. The growth of individual beta-amyloid protofibrils on a mica substrate was followed over several hours. The first in situ visualization of protofibril formation from single aggregate units of beta amyloid is reported. The growth of these protofibrils through the subsequent addition of these aggregate units is also observed. Growth of the protofibrils is bi-directional and the outgrowth of protofibrils from a common amyloid/heterogeneous core is also observed. Elongation also occurred by the addition of protofibrils from solution. This data provides an exciting insight into the early stages of beta-amyloid fibrillization and can be used to enhance the understanding of the mechanism(s) by which beta-amyloid fibrillizes and may consequently enable inhibition of one or more stages of fibrillization as a potential therapeutic strategy. PMID- 10801353 TI - Molecular modelling study of HIV p17gag (MA) protein shell utilising data from electron microscopy and X-ray crystallography. AB - The matrix protein p17gag (MA) is a product of proteolytic cleavage of the gag gene encoded polyprotein (pr55gag) and is formed when HIV particles undergo the process of maturation. The MA protein is associated with the inner surface of the viral membrane and determines the overall shape of the virion. Previous studies have shown the existence of trimers of MA in solution and in the crystalline state. Here, we used molecular modelling methods to identify feasible interactions between pairs of MA trimers and have related this to structural data from electron microscopy. A systematic search docking procedure was able to identify many energetically favourable conformations for a pair of trimers, including some which have been previously reported. These conformations were used to generate several networks of MA trimers, which were then evaluated against structural observations of the MA network. The model suggested here provides a good match with experimental data such as the spacing between gag protein rings, the number and disposition of glycoprotein (gp41-gp120) knobs and the number of copies of MA in a virus particle. It also rationalizes the observed distribution of sizes of virus particles and is consistent with the presence of icosahedral organisation in mature HIV. Energy minimisation performed with explicit water and counter ions, was used to identify residues participating in inter-trimer interactions. The nature of these interactions is discussed in relation to the conservation of these residues in reported variants of the HIV and SIV MA protein sequences. PMID- 10801354 TI - Crystal structure of a plant ribonuclease, RNase LE. AB - Ribonuclease LE (RNase LE) from cultured tomato (Lycopersicon esculentum) cells is a member of the RNase T(2) family showing broad base specificity. The crystal structure of RNase LE has been determined at 1.65 A resolution. The structure consists of seven alpha-helices and seven beta-strands, belonging to an alpha+beta type structure. Comparison of the structure of RNase LE with that of RNase Rh, a microbial RNase belonging to the RNase T(2) family, reveals that while the overall folding topologies are similar to each other, major insertions and deletions are found at the N-terminal regions. The structural comparison, an amino acid sequence alignment of the RNase T(2) enzymes, and comparison of the disulfide-bonding pattern of these enzymes show that the structure of RNase LE shown here is the basic framework of the animal/plant subfamily of RNase T(2) enzymes (including a self-incompatibility protein called S-RNase), and the structure of RNase Rh is that of the fungal subfamily of RNase T(2) enzymes (including RNase T(2)). Subsequently, we superposed the active-site of the RNase LE with that of RNase Rh and found that (1) His39, Trp42, His92, Glu93, Lys96, and His97 of RNase LE coincided exactly with His46, Trp49, His104, Glu105, Lys108, and His109, respectively, of RNase Rh, and (2) two conserved water molecules were found at the putative P(1) sites of both enzymes. These facts suggest that plant RNase LE has a very similar hydrolysis mechanism to that of fungal RNase Rh, and almost all the RNase T(2) enzymes widely distributed in various species share a common catalytic mechanism. A cluster of hydrophobic residues was found on the active-site face of the RNase LE molecule and two large hydrophobic pockets exist. These hydrophobic pockets appear to be base binding sites mainly by hydrophobic interactions and are responsible for the base non specificity of RNase LE. PMID- 10801355 TI - Crystal structures of Toxoplasma gondii adenosine kinase reveal a novel catalytic mechanism and prodrug binding. AB - Adenosine kinase (AK) is a key purine metabolic enzyme from the opportunistic parasitic protozoan Toxoplasma gondii and belongs to the family of carbohydrate kinases that includes ribokinase. To understand the catalytic mechanism of AK, we determined the structures of the T. gondii apo AK, AK:adenosine complex and the AK:adenosine:AMP-PCP complex to 2.55 A, 2.50 A and 1.71 A resolution, respectively. These structures reveal a novel catalytic mechanism that involves an adenosine-induced domain rotation of 30 degrees and a newly described anion hole (DTXGAGD), requiring a helix-to-coil conformational change that is induced by ATP binding. Nucleotide binding also evokes a coil-to-helix transition that completes the formation of the ATP binding pocket. A conserved dipeptide, Gly68 Gly69, which is located at the bottom of the adenosine-binding site, functions as the switch for domain rotation. The synergistic structural changes that occur upon substrate binding sequester the adenosine and the ATP gamma phosphate from solvent and optimally position the substrates for catalysis. Finally, the 1.84 A resolution structure of an AK:7-iodotubercidin:AMP-PCP complex reveals the basis for the higher affinity binding of this prodrug over adenosine and thus provides a scaffold for the design of new inhibitors and subversive substrates that target the T. gondii AK. PMID- 10801356 TI - The structure of an insect chymotrypsin. AB - The South American imported fire ant (Solenopsis invicta), without natural enemies in the United States, widely infests the southern United States, causing more than a half billion dollars in health and agriculture-related damage annually in Texas alone. Fire ants are resistant to most insecticides, so control will require a more fundamental understanding of their biochemistry and metabolism leading to the design of selective, ecologically safe insecticides. The 4th instar larvae play a crucial role in the nutrition of the colony by secreting proteinases (especially chymotrypsin) which digest food products for the entire colony. The first structure of an ant proteolytic enzyme, fire ant chymotrypsin, was determined to atomic resolution (1.7 A). A structural comparison of the ant and mammalian structures confirms the "universality" of the serine proteinase motif and reveals a difference at residues 147-148, which are proteolytically removed in the bovine enzyme but are firmly intact in the ant chymotrypsin, suggesting a different activation mechanism for the latter. Likewise, the absence of the covalently attached propeptide domain (1-15) further suggests an uncharacteristic activation mechanism. The presence of Gly189 in the S1 site is an atypical feature of this chymotrypsin and is comparable only to human leukocyte elastase, hornet chymotrypsin and fiddler crab collagenase. Binding studies confirm the chymotrypsin nature of this novel enzyme. PMID- 10801357 TI - The transition between the open and closed states of rubisco is triggered by the inter-phosphate distance of the bound bisphosphate. AB - d-Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) catalyses the central CO(2)-fixing reaction of photosynthesis in a complex, multiple-step process. Several structures of rubisco complexed with substrate analogues, inhibitors and products have been determined by X-ray crystallography. The structures fall into two well-defined and distinct states. The active site is either "open" or "closed". The timing and mechanism of the transition between these two states have been uncertain. We solved the crystal structure of unactivated (metal-free) rubisco from tobacco with only inorganic phosphate bound and conclude that phosphate binding per se does not trigger closure, as it does in the similarly structured enzyme, triosephosphate isomerase. Comparison of all available rubisco structures suggests that, instead, the distance between the terminal phosphates (P1 and P2) of the bisphosphate ligand is the trigger: if that distance is less than 9.1 A, then the active site closes; if it is greater than 9.4 A then the enzyme remains open. Shortening of the inter-phosphate distance results from the ligand binding in a more curved conformation when O atoms of the ligand's sugar backbone interact either with the metal, if it is present, or with charged groups in the metal-binding site, if the metal is absent. This shortening brings the P1 phosphate into hydrogen bonding contact with Thr65. Thr65 exists in two discrete states related by a rotation of the backbone psi torsion angle. This rotation is coupled to domain rotation and hence to active site closure. Rotation of the side chain of Thr65 also affects the C-terminal strand of large subunit which packs against Loop 6 after closure. The position of the C-terminal strand in the closed state is stabilised by multiple polar interactions with a distinctive highly charged latch site involving the side-chain of Asp473. In the open state, this latch site may be occupied instead by phosphorylated anions. PMID- 10801358 TI - Stabilization of GroEL minichaperones by core and surface mutations. AB - We report the crystal structures of two hexa-substituted mutants of a GroEL minichaperone that are more stable than wild-type by 7.0 and 6.1 kcal mol(-1). Their structures imply that the increased stability results from multiple factors including improved hydrophobic packing, optimised hydrogen bonding and favourable structural rearrangements. It is commonly believed that protein core residues are immutable and generally optimized for energy, while on the contrary, surface residues are variable and hence unimportant for stability. But, it is now becoming clear that mutations of both core and surface residues can increase protein stability, and that protein cores are more flexible and thus more tolerant to mutation than expected. Sequence comparison of homologous proteins has provided a way to pinpoint the residues that contribute constructively to stability and to guide the engineering of protein stability. Stabilizing mutations identified by this approach are most frequently located at protein surfaces but with a few found in protein cores. In the latter case, local flexibility in the hydrophobic core is the key factor that allows the energetically favourable burial of larger hydrophobic side-chains without undue energetic penalties from steric clashes. PMID- 10801359 TI - De novo determination of protein structure by NMR using orientational and long range order restraints. AB - Orientational and novel long-range order restraints available from paramagnetic systems have been used to determine the backbone solution structure of the cytochrome c' protein to atomic resolution in the complete absence of restraints derived from the nuclear Overhauser effect. By exploiting the complementary geometric dependence of paramagnetic pseudocontact shifts and the recently proposed Curie-dipolar cross correlated relaxation effect, in combination with orientational constraints derived from residual dipolar coupling, autorelaxation rate ratios and secondary structure constraints, it is possible to define uniquely the fold and refine the tertiary structure of the protein (0.73 A backbone rmsd for 82/129 amino acid residues) starting from random atomic Cartesian coordinates. The structure calculation protocol, developed using specific models to describe the novel constraint interactions, is robust, requiring no precise a priori estimation of the various interaction strengths, and provides unambiguous convergence based only on the value of the target function. Tensor eigenvalues and their component orientations are allowed to float freely, and are thus simultaneously determined, and found to converge, during the structure calculation. PMID- 10801360 TI - Topological and energetic factors: what determines the structural details of the transition state ensemble and "en-route" intermediates for protein folding? An investigation for small globular proteins. AB - Recent experimental results suggest that the native fold, or topology, plays a primary role in determining the structure of the transition state ensemble, at least for small, fast-folding proteins. To investigate the extent of the topological control of the folding process, we studied the folding of simplified models of five small globular proteins constructed using a Go-like potential to retain the information about the native structures but drastically reduce the energetic frustration and energetic heterogeneity among residue-residue native interactions. By comparing the structure of the transition state ensemble (experimentally determined by Phi-values) and of the intermediates with those obtained using our models, we show that these energetically unfrustrated models can reproduce the global experimentally known features of the transition state ensembles and "en-route" intermediates, at least for the analyzed proteins. This result clearly indicates that, as long as the protein sequence is sufficiently minimally frustrated, topology plays a central role in determining the folding mechanism. PMID- 10801361 TI - Coupled kinetic traps in cytochrome c folding: His-heme misligation and proline isomerization. AB - The effect of His-heme misligation on folding has been investigated for a triple mutant of yeast iso-2 cytochrome c (N26H,H33N,H39K iso-2). The variant contains a single misligating His residue at position 26, a location at which His residues are found in several cytochrome c homologues, including horse, tuna, and yeast iso-1. The amplitude for fast phase folding exhibits a strong initial pH dependence. For GdnHCl unfolded protein at an initial pH<5, the observed refolding at final pH 6 is dominated by a fast phase (tau(2f)=20 ms, alpha(2f)=90 %) that represents folding in the absence of misligation. For unfolded protein at initial pH 6, folding at final pH 6 occurs in a fast phase of reduced amplitude (alpha(2f) approximately 20 %) but the same rate (tau(2f)=20 ms), and in two slower phases (tau(m)=6-8 seconds, alpha(m) approximately 45 %; and tau(1b)=16-20 seconds, alpha(1b) approximately 35 %). Double jump experiments show that the initial pH dependence of the folding amplitudes results from a slow pH-dependent equilibrium between fast and slow folding species present in the unfolded protein. The slow equilibrium arises from coupling of the His protonation equilibrium to His-heme misligation and proline isomerization. Specifically, Pro25 is predominantly in trans in the unligated low-pH unfolded protein, but is constrained in a non-native cis isomerization state by His26-heme misligation near neutral pH. Refolding from the misligated unfolded form proceeds slowly due to the large energetic barrier required for proline isomerization and displacement of the misligated His26-heme ligand. PMID- 10801362 TI - A breakdown of symmetry in the folding transition state of protein L. AB - The 62 residue IgG binding domain of protein L consists of a central alpha-helix packed on a four-stranded beta-sheet formed by N and C-terminal beta-hairpins. The overall topology of the protein is quite symmetric: the beta-hairpins have similar lengths and make very similar interactions with the central helix. Characterization of the effects of 70 point mutations distributed throughout the protein on the kinetics of folding and unfolding reveals that this symmetry is completely broken during folding; the first beta-hairpin is largely structured while the second beta-hairpin and helix are largely disrupted in the folding transition state ensemble. The results are not consistent with a "hydrophobic core first" picture of protein folding; the first beta-hairpin appears to be at least as ordered at the rate limiting step in folding as the hydrophobic core. PMID- 10801364 TI - Thank you from JMB! JMB is indebted to the following people for their help and guidance in 1999: PMID- 10801365 TI - Cochrane collaborative review group on peripheral vascular diseases: review abstracts PMID- 10801363 TI - Local and long-range interactions in the molten globule state: A study of chimeric proteins of bovine and human alpha-lactalbumin. AB - The molten globule state of alpha-lactalbumin has ordered secondary structure in the alpha-domain, which comprises residues 1 to 34 and 86 to 123. In order to investigate which part of a polypeptide is important for stabilizing the molten globule state of alpha-lactalbumin, we have produced and studied three chimeric proteins of bovine and human alpha-lactalbumin. The stability of the molten globule state formed by domain-exchanged alpha-lactalbumin, in which the amino acid sequence in the alpha-domain comes from human alpha-lactalbumin and that in the beta-domain comes from bovine alpha-lactalbumin, is the same as that of human alpha-lactalbumin and is substantially greater than that of bovine alpha lactalbumin. Therefore, our results show that the stability of the molten globule state of alpha-lactalbumin is determined by the alpha-domain and the beta-domain is not important for stabilizing the molten globule state. The substitution of residues 1 to 34 of bovine alpha-lactalbumin with those of human alpha lactalbumin substantially increases the stability of the molten globule state, while the substitution of residues 86 to 123 of bovine alpha-lactalbumin with those of human alpha-lactalbumin decreases the stability of the molten globule state. Therefore, residues 1 to 34 in human alpha-lactalbumin is more important for the stability of the human alpha-lactalbumin molten globule state than residues 86 to 123. The stabilization of the molten globule state due to substitution of both residues 1 to 34 and 86 to 123 is not identical with the sum of the two individual substitutions, demonstrating the non-additivity of the stabilization of the molten globule state. This result indicates that there is a long-range interaction between residues 1 to 34 and 86 to 123 in the molten globule state of human alpha-lactalbumin. The differences in the stabilities of the molten globule states are well correlated with the averaged helical propensity values in the alpha-domain when the long-range interactions are negligible, suggesting that the local interaction is the dominant term for determining the stability of the molten globule state. Our results also indicate that the apparent cooperativity is closely linked to the stability of the molten globule state, even if the molten globule state is weakly cooperative. PMID- 10801366 TI - Intimal hyperplasia in vascular grafts. PMID- 10801367 TI - Projected workload for a vascular service in 2020. AB - OBJECTIVE: to estimate the workload of a vascular service during the next two decades as the proportion of people aged over 65 years increases. METHODS: the study used the vascular registry data of Tampere University Hospital and the population data of Pirkanmaa region provided by the Central Statistical Office in Finland. The current workload is 1420 vascular procedures per million inhabitants yearly (951 surgical and 207 endovascular). Sixty-five per cent of all procedures are done on people over 65 years old. Pirkanmaa has a population of 440 000 persons of whom 15.6% are over 65 years. According to the population data the population will increase to 460 000 persons by the year 2020 and 22.9% of them will be over 65 years old. RESULTS: The total amount of procedures will rise by 40.5% (1906) and the increase in endovascular and surgical group will be 39.2% (640) and 43.5% (1265) respectively. The proportion of treated patients over 65 years will rise from 65.0% to 70.5%. In the next two decades the amount of patients with claudication will increase by 35.4%, critical limb ischaemia by 44.2%, carotid surgery by 34.0%, abdominal aortic aneurysms by 40.7%, acute limb ischaemia by 45.0% and access surgery by 27.4%. CONCLUSION: In the next two decades the number of elderly people will increase so rapidly that, whatever happens to the incidence and prevalence of peripheral vascular disease, the workload for a vascular service will increase significantly. PMID- 10801368 TI - Carotid atherosclerosis in relation to symptoms from the territory supplied by the carotid artery. AB - OBJECTIVES: To investigate the relationship between intima-media thickness in the common carotid artery, plaque morphology in the carotid bifurcation and symptoms of cerebral embolism. Design prospective study of consecutive patients referred for carotid duplex examination. METHODS: One hundred and eighty-eight patients were classified by one of two neurologists into four categories (symptomatic, asymptomatic, undefined, uncertain). Carotid atherosclerosis was measured by means of high-resolution ultrasound technique. Carotid plaques were classified based on visual evaluation of plaque echogenicity. RESULTS: Intima-media thickness (IMT) was correlated to presence of plaques, age and gender, but not to symptoms referable to the carotid circulation. Carotid plaques were more common in symptomatic than in asymptomatic vessels (p<0.05). There was no difference in plaque occurrence between the ipsi- and contralateral sides in the symptomatic patients, neither in frequency of echolucent plaques between the sides or between symptomatic and asymptomatic patients. Echogenic plaques were more common and generally smaller than echolucent plaques. CONCLUSIONS: IMT correlates with presence of plaques, but not with symptoms from the territory supplied by the carotid artery. Echolucent plaques in the carotid bifurcation are not more frequently symptomatic than echogenic plaques are, in patients with low-to moderate degree of carotid stenosis. PMID- 10801369 TI - A regional collaborative audit of the practice and outcome of carotid endarterectomy in the United Kingdom. Northern Regional Carotid Endarterectomy Audit Group. AB - OBJECTIVES: To evaluate performance and outcome of carotid endarterectomy (CEA) against agreed audit standards within one English health region. Design a prospective collaborative audit over twelve months (November 1994 to October 1995) involving all surgeons undertaking CEA within one English health region. METHODS: Audit standards were agreed by all participating surgeons at the outset based on existing national guidelines. Data were abstracted from clinical notes. Outcomes were reviewed by clinicians 30 days post-surgery. A confidential individualised report of the results was provided to each surgeon. A survey of participating surgeons sought to evaluate the audit process. RESULTS: Ten surgeons performed 139 CEAs on 134 individuals (64% men). Median per surgeon was 12 (range 1-44). Audit standards were generally achieved: 114 (82%) patients had symptomatic carotid stenosis of 70-99%, 14 (10%) were asymptomatic. The median time from first referral to hospital to operation was 4.8 months (interquartile range 3.0-7.3). The rate of disabling stroke or death at 30 days was 2.2% (95% confidence interval (CI) 0.4-6.4%). Surgeons valued the audit. CONCLUSIONS: The study showed that in the study area CEA was performed predominantly on high-risk patients with low subsequent surgical mortality. PMID- 10801370 TI - Antiplatelet therapy and other interventions after revascularisation procedures in patients with peripheral arterial disease: a meta-analysis. AB - OBJECTIVES: To evaluate the efficacy of conservative adjuvant therapy after revascularisation procedures in patients with peripheral arterial disease. DESIGN: meta-analysis. MATERIALS: English-language studies published from 1976 to 1997. METHODS: Reports on conservative therapies in patients with peripheral arterial disease after percutaneous transluminal angioplasty, endarterectomy, thromboendarterectomy or bypass grafting were eligible. Uncontrolled or retrospective studies, double reports or trials without clinical outcomes were excluded. Included studies were graded as level 1 (randomised and double- or assessor-blind), level 2 (open randomised), or level 3 (non-randomised comparative). (Loss of) patency, amputation, vascular events and mortality were the outcomes considered. When feasible, end-of-treatment results, either continuous or binary, were combined with appropriate statistical methods. RESULTS: Thirty-two studies were included. Compared to non-active control, aspirin with dipyridamole improved (loss of) patency (odds ratio (OR) 0.69, 95% confidence interval (CI), 0.53 to 0.90) and mortality (OR 0.80, 95% CI, 0.57 to 1.14); ticlopidine improved (loss of) patency (OR 0.53, 95% CI, 0.33 to 0.85) and amputation (OR 0.29, 95% CI, 0.08 to 1.01). Data on the effectiveness of vitamin K inhibitors were not conclusive. CONCLUSIONS: Patients with peripheral arterial disease improve their outcome by receiving aspirin with dipyridamole or ticlopidine after a revascularisation procedure. PMID- 10801371 TI - Tissue engineering of vascular grafts: human cell seeding of decellularised porcine matrix. AB - OBJECTIVES: To develop a biocompatible and mechanically stable vascular graft combining human cells and a xenogenic acellular matrix. DESIGN/MATERIALS: Decellularised matrix tubes were obtained by enzymatic cell extraction of native porcine aortas. Endothelial cells and myofibroblasts were isolated from human saphenous veins and grown in cell cultures. The inner surface of the tubes was seeded with endothelial cells or myofibroblasts and exposed to pulsatile flow. RESULTS: After cell extraction, the absence of cellular components, as well as the maintenance of matrix integrity, was demonstrated by means of light microscopy and scanning electron microscopy. Furthermore, the porcine matrix was successfully seeded with human endothelial cells, which grew to a monolayer under flow conditions. Stable biomechanical properties were achieved at physiological perfusion pressures in vitro. CONCLUSIONS: Cellular components can be extracted from native porcine blood vessels. Vascular grafts can be generated in vitro of animal acellular matrix and human cells. PMID- 10801372 TI - Differential responses in human subcutaneous and skeletal muscle vascular beds to critical limb ischaemia. AB - OBJECTIVES: To investigate the effects of chronic ischaemia on the subcutaneous and the skeletal muscle resistance vasculature. To understand the redistribution of available blood in the ischaemic limb. METHODS: Human subcutaneous and skeletal muscle resistance arteries were obtained from limbs amputated for critical limb ischaemia and studied under isobaric conditions using pressure myography. Morphological measurements of wall and lumen were analysed using light microscopy and image analysis. Vasoconstrictor responses to potassium and adrenoceptor agonists were used to measure functional status. Noradrenaline re uptake mechanisms and alpha(1)-selectivity were investigated. RESULTS: Both human skeletal muscle and subcutaneous resistance arteries undergo a severe atrophy of the arterial wall in ischaemic conditions. However, whereas subcutaneous resistance arteries become less able to vasoconstrict to adrenoceptor stimulation, the response of skeletal muscle resistance arteries becomes exaggerated and significantly augmented. This is true in response to both the endogenous vasoconstrictor noradrenaline and the alpha(1)-selective adrenoceptor agonist phenylephrine. CONCLUSIONS: Hypersensitivity to circulating catecholamines in the skeletal muscle vascular resistance bed may contribute to the progression of ischaemic disease by differentially diverting available blood to the subcutaneous tissue to the detriment of skeletal muscle perfusion. PMID- 10801373 TI - Prevention of renal impairment following aortic cross-clamping by manipulation of the endogenous renal nitric oxide response. AB - OBJECTIVE: Infrarenal aortic cross-clamp-induced lower torso ischaemia reperfusion injury is associated with impairment of glomerular filtration rate and upregulation of endogenous renal nitric oxide production. The aim of this study was to investigate whether manipulation of the endogenous renal nitric oxide response can ameliorate subsequent renal injury. METHODS: Groups of male Wistar rats (n = 6) were treated with one of the following agents before being subjected to 60 min of infrarenal aortic cross-clamping: saline (control), L-NMMA (a pan nitric oxide synthase inhibitor), 1400W (a highly selective iNOS inhibitor), hydrocortisone (an inhibitor of the systemic inflammatory response), L-arginine (the substrate for nitric oxide synthase) and NOC-18 (a nitric oxide donor). Animals were recovered after a left nephrectomy. The glomerular filtration rate (GFR) of the remaining kidney was measured on the second and seventh postoperative day using a 99Tc DPTA clearance technique as an index of renal injury. RESULTS: Animals treated with L-NMMA prior to aortic cross-clamping had a significantly impaired GFR compared to controls on the second (p<0. 01, Mann-Whitney U -test) and seventh (p<0.05, Mann-Whitney U -test) postoperative day. Hydrocortisone and 1400W had no significant effect on GFR on the second or seventh postoperative day. L-arginine and NOC-18 had no influence on GFR on the second postoperative day but significantly improved GFR on the seventh postoperative day. CONCLUSIONS: These results show that the endogenous nitric oxide response protects the kidneys from ischaemia-reperfusion injury. Manipulation of the renal nitric oxide response may have therapeutic benefits to patients undergoing aortic aneurysm repair by preventing acute renal failure. PMID- 10801374 TI - The surgical management of renovascular hypertension in children and young adults. AB - OBJECTIVES: To assess the outcome and durability of operative revascularisation in young patients with renovascular hypertension. DESIGN: Retrospective study. METHOD: The records of all young patients (under 25 years) operated on for renovascular hypertension at St Mary's Hospital 1988-1998 were reviewed. We assessed the aetiology of hypertension, operations performed, effect of treatment on blood pressure, renal function and requirement for antihypertensive medication during follow-up. RESULTS: Ten patients were identified who had been considered for surgery, of median age 16 years (22 months to 22 years). Fibromuscular dysplasia was present in five patients, mid-aortic syndrome (MAS) in four and neurofibromatosis in one. Operations performed were aortorenal bypass (three), aorto-aortic bypass+/-renal bypass (three), splenorenal bypass (one) and autotransplantation (one). Of the three patients treated by balloon angioplasty, only one had a successful result. One patient with MAS is currently awaiting surgery. Over a median follow-up of 24 months (8-144), seven patients are normotensive off all antihypertensive medication. Of two patients on reduced doses of medication, one (splenorenal bypass) required surgical repair of a late (9 years) coeliac stenosis. CONCLUSIONS: The surgical treatment of renovascular hypertension in carefully selected young patients gives durable results. Blood pressure is well controlled long-term, and the need for antihypertensive medication is removed altogether in the majority of patients. PMID- 10801375 TI - Placement of a fenestrated Palmaz stent across the renal arteries. Feasibility and outcome in an animal study. AB - OBJECTIVES: To investigate the feasibility of placing stents across renal arteries. Design we have studied in pigs: (i) the feasibility of accurately placing a fenestrated stent in front of one renal ostium; (ii) the short-term effects on renal arteries and function after the placement of such a fenestrated stent. MATERIALS AND METHODS: Eight fenestrated Palmaz stents were placed over pigs' renal ostia under fluoroscopy. Five weeks later, angiograms were performed and the animals were sacrificed. Proliferation of the healing tissues over the ostia was measured and analysed by microscopy. Serum creatinine was measured prior to all angiograms and at 5 weeks. RESULTS: All eight stents were correctly placed. One stent later migrated and was excluded from the study. One pig died at day 1. Gross examination confirmed the correct placement of the fenestrations in four pigs out of seven (57%). In the six remaining pigs, at 5 weeks, there was no angiographic evidence of stent misplacement and all the kidneys were fully perfused. Nine renal ostia were covered by struts and neointima with a mean area of coverage of 38+/-5% altogether. No tissue proliferation was observed over the three renal ostia located in front of the fenestration. Serum creatinine did not significantly increase at 5 weeks. CONCLUSION: Creating a fenestration in a stent for renal arteries may be worth while in order to avoid neointimal covering of the renal ostia. However, accurate placement of such a fenestrated stent remains a difficult task. PMID- 10801376 TI - Relationship between intrathecal oxygen tension and ultrastructural changes in the spinal cord during experimental aortic clamping. AB - OBJECTIVES: To investigate spinal cord ultrastructure related to cerebrospinal fluid (CSF) oxygenation. DESIGN: experimental aortic occlusion model with intrathecal oxygen tension monitoring. MATERIALS AND METHODS: Two groups of pigs underwent proximal (P) or double (D) aortic occlusion for 30 min followed by 1 h of reperfusion. In a third group (I) segmental arteries distal to T3 were clamped for 90 min. A thin pO(2), pCO(2) and pH sensor was placed intrathecally for continuous monitoring of CSF. Spinal cord segments were studied by electron microscopy (EM). RESULTS: In group P, CSF-pO(2)rapidly decreased during clamping and major changes in pH and pCO(2)were seen. EM demonstrated neuronal degeneration with loss of cellular integrity and severe affection of organelles. In the group D, CSF oxygenation decreased to about half, but with only moderate changes in the metabolic parameters. Group I showed no significant changes in CSF measurements. The latter groups were similar at EM, showing only mild mitochondrial changes. CONCLUSIONS: The level of CSF oxygenation during aortic cross-clamping or segmental artery interruption seems to correlate with ultrastructural changes in the spinal cord. This online intrathecal monitoring technique may provide valuable information on spinal cord circulation during thoracoabdominal aortic surgery. PMID- 10801377 TI - Freedom from endoleak after endovascular aneurysm repair does not equal treatment success. AB - OBJECTIVE: To determine whether freedom from endoleak after endovascular repair of abdominal aortic aneurysm (EVAR) is a reliable guide to freedom from persistent or recurrent pressurisation of the aneurysm sac (endotension) and therefore freedom from risk of rupture. PATIENTS AND METHODS: The records of 55 patients followed for more than 3 months after EVAR were reviewed to correlate the presence or absence of endoleak on contrast-enhanced CT and/or angiography with changes in maximum aneurysm diameter (DMAX). RESULTS: in 22 (40%) patients there was no significant change in DMAX during follow-up. In 21 of these no endoleak was observed on CT or angiography. One patient developed a secondary side-branch endoleak which remains under observation. In 18 (33%) patients, DMAX decreased during follow-up. Thirteen of these remained free of endoleak. Four patients developed secondary endoleaks which were treated by secondary intervention. One patient with persistent primary endoleak suffered fatal aneurysm rupture three days before planned intervention. DMAX increased in 15 (27%) patients. In only five of these could an endoleak be identified on CT and/or angiography. One primary side-branch endoleak persists following failed embolisation. Four secondary endoleaks have been corrected by secondary intervention. Four of the remaining 10 patients died suddenly from unknown cause. All had DMAX greater than 65 mm at last follow-up. One patient underwent late conversion, which suggested continued pressurisation through thrombus at the site of a "sealed" primary proximal endoleak. Two patients are scheduled to undergo embolisation of patent side-branches revealed only by Levovist enhanced Duplex scanning and three patients remain under observation. CONCLUSION: Freedom from endoleak on conventional imaging incorrectly suggested freedom from endotension in 10 (18%) of our patients. Follow-up after endovascular repair must include regular measurement of DMAX and/or aneurysm sac volume to identify those patients who remain at risk of rupture. PMID- 10801378 TI - Extra-anatomical long-saphenous conduit for decompression of superior vena cava syndrome. PMID- 10801379 TI - Use of a bifurcated graft for endovascular repair of a common iliac aneurysm after previous open aortic aneurysm surgery. PMID- 10801380 TI - Ruptured aneurysms of superficial femoral artery. PMID- 10801381 TI - Is bacterial translocation a cause of aortic graft sepsis? PMID- 10801382 TI - Delayed rupture of non-anastomotic aneurysm of an infected PTFE graft following femoropopliteal bypass surgery. PMID- 10801383 TI - Gradual Hunterian ligation for infected prosthetic bypass. AB - OBJECTIVE: To review gradual snare occlusion for the management of complex or recurrent graft infection. PATIENTS AND METHODS: Medical records of patients treated with gradual snare occlusion following graft infection were reviewed for indication for operation, type of bypass and graft material used. In addition, infecting organism, grade of infection (Szilagyi) and outcome were recorded. RESULTS: Four femoropopliteal, two extra-anatomic (axillofemoral) and aortobifemoral bypasses were included in this study. All had chronic infection (Szilagyi grade III) with onset of 4 to 24 months and two of which were recurrent. The causative organisms were coagulase-negative staphylococci, Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus in three patients, with no organism isolated in the remaining cases. There was no loss of limb following gradual snare occlusion but there was only one death due to aortic stump rupture 2 weeks later. CONCLUSION: Gradual snare occlusion is an alternative for the management of chronic or recurrent graft infection. PMID- 10801384 TI - Tuberculous false aneurysm of the femoral artery managed by endoluminal stent graft insertion. PMID- 10801385 TI - Parental origin and phenotype of triploidy in spontaneous abortions: predominance of diandry and association with the partial hydatidiform mole. AB - The origin of human triploidy is controversial. Early cytogenetic studies found the majority of cases to be paternal in origin; however, recent molecular analyses have challenged these findings, suggesting that digynic triploidy is the most common source of triploidy. To resolve this dispute, we examined 91 cases of human triploid spontaneous abortions to (1) determine the mechanism of origin of the additional haploid set, and (2) assess the effect of origin on the phenotype of the conceptus. Our results indicate that the majority of cases were diandric in origin because of dispermy, whereas the maternally-derived cases mainly originated through errors in meiosis II. Furthermore, our results indicate a complex relationship between phenotype and parental origin: paternally-derived cases predominate among "typical" spontaneous abortions, whereas maternally derived cases are associated with either early embryonic demise or with relatively late demise involving a well-formed fetus. As the cytogenetic studies relied on analyses of the former type of material and the molecular studies on the latter sources, the discrepancies between the data sets are explained by differences in ascertainment. In studies correlating the origin of the extra haploid set with histological phenotype, we observed an association between paternal-but not maternal-triploidy and the development of partial hydatidiform moles. However, only a proportion of paternally derived cases developed a partial molar phenotype, indicating that the mere presence of two paternal genomes is not sufficient for molar development. PMID- 10801386 TI - Localization of psoriasis-susceptibility locus PSORS1 to a 60-kb interval telomeric to HLA-C. AB - Recent genome scans have established the presence of a major psoriasis susceptibility locus in the human leukocyte antigen (HLA) complex on chromosome 6p21.3. To narrow the interval for candidate gene testing, we performed a linkage disequilibrium analysis of 339 families, with the use of 62 physically mapped microsatellite markers spanning the major histocompatibility complex (MHC). As detected by use of the transmission/disequilibrium test (TDT), individual markers yielded significant linkage disequilibrium across most of the MHC. However, the strongest evidence for marker-trait disequilibrium was found in an approximately 300-kb region extending from the MICA gene to the corneodesmosin gene. Maximum likelihood haplotypes were constructed across the entire MHC in the original sample and across a 1.2-Mb region of the central MHC in an expanded sample containing 139 additional families. Short (two- to five-marker) haplotypes were subjected to the TDT using a "moving-window" strategy that reduced the variability of TDT P values relative to the single-locus results. Furthermore, the expanded sample yielded a sharp peak of evidence for linkage disequilibrium that spanned approximately 170 kb and that was centered 100 kb telomeric to HLA C. The 1.2-Mb interval was further dissected by means of recombinant ancestral haplotype analysis. This analysis identified risk haplotype 1 (RH1), which is a 60-kb fragment of ancestral haplotype 57.1, on all identifiable HLA risk haplotypes. One of these haplotypes exhibits significant linkage disequilibrium with psoriasis but does not carry Cw6, which is the HLA allele most strongly associated with the disease. These results demonstrate that RH1 is highly likely to carry the disease allele at PSORS1, and they exclude HLA-C and corneodesmosin with a high degree of confidence. PMID- 10801387 TI - Characterization of human crossover interference. AB - We present an analysis of crossover interference over the entire human genome, on the basis of genotype data from more than 8,000 polymorphisms in eight CEPH families. Overwhelming evidence was found for strong positive crossover interference, with average strength lying between the levels of interference implied by the Kosambi and Carter-Falconer map functions. Five mathematical models of interference were evaluated: the gamma model and four versions of the count-location model. The gamma model fit the data far better than did any of the other four models. Analysis of intercrossover distances was greatly superior to the analysis of crossover counts, in both demonstrating interference and distinguishing between the five models. In contrast to earlier suggestions, interference was found to continue uninterrupted across the centromeres. No convincing differences in the levels of interference were found between the sexes or among chromosomes; however, we did detect possible individual variation in interference among the eight mothers. Finally, we present an equation that provides the probability of the occurrence of a double crossover between two nonrecombinant, informative polymorphisms. PMID- 10801388 TI - The power of genomic control. AB - Although association analysis is a useful tool for uncovering the genetic underpinnings of complex traits, its utility is diminished by population substructure, which can produce spurious association between phenotype and genotype within population-based samples. Because family-based designs are robust against substructure, they have risen to the fore of association analysis. Yet, if population substructure could be ignored, this robustness can come at the price of power. Unfortunately it is rarely evident when population substructure can be ignored. Devlin and Roeder recently have proposed a method, termed "genomic control" (GC), which has the robustness of family-based designs even though it uses population-based data. GC uses the genome itself to determine appropriate corrections for population-based association tests. Using the GC method, we contrast the power of two study designs, family trios (i.e., father, mother, and affected progeny) versus case-control. For analysis of trios, we use the TDT test. When population substructure is absent, we find GC is always more powerful than TDT; furthermore, contrary to previous results, we show that as a disease becomes more prevalent the discrepancy in power becomes more extreme. When population substructure is present, however, the results are more complex: TDT is more powerful when population substructure is substantial, and GC is more powerful otherwise. We also explore general issues of power and implementation of GC within the case-control setting and find that, economically, GC is at least comparable to and often less expensive than family-based methods. Therefore, GC methods should prove a useful complement to family-based methods for the genetic analysis of complex traits. PMID- 10801389 TI - High frequency of cystic fibrosis transmembrane regulator mutation L997F in patients with recurrent idiopathic pancreatitis and in newborns with hypertrypsinemia. PMID- 10801390 TI - On the age of the most prevalent Gaucher disease-causing mutation, N370S. PMID- 10801391 TI - Evolution of regulatory responses to feeding in snakes. AB - Do animal species that normally consume large meals at long intervals evolve to down-regulate their metabolic physiology while fasting and to up-regulate it steeply on feeding? To test this hypothesis, we compared postfeeding regulatory responses in eight snake species: four frequent feeders on small meals and four infrequent feeders on large meals. For each species, we measured factorial changes in metabolic rate, in activities and capacities of five small intestinal brush border nutrient transporters, and in masses of eight organs that function in nutrient processing after consumption of a rodent meal equivalent to 25% of the snake's body mass. It turned out that, compared with frequent feeders, infrequent feeders digest that meal more slowly; have lower metabolic rates, organ masses, and nutrient uptake rates and capacities while fasting; have higher energy expenditure during digestion; and have higher postfeeding factorial increases in metabolic rate, organ masses, and nutrient uptake rates and capacities. These conclusions, which conform to the hypothesis mentioned above, remain after phylogeny has been taken into account. The small organ masses and low nutrient transporter activities during fasting contribute to the low fasting metabolism of infrequent feeders. Quantitative calculations of partial energy budgets suggest that energy savings drive the evolution of low mass and activities of organs during fasting and of large postfeeding regulatory responses in infrequent feeders. We propose further tests of this hypothesis among other snake species and among other ectotherms. PMID- 10801392 TI - Effect of ontogenetic increases in body size on burst swimming performance in tadpoles of the striped marsh frog, Limnodynastes peronii. AB - The effect of ontogenetic increases in total length on burst swimming performance was investigated in tadpoles of the striped marsh frog (Limnodynastes peronii) over the total-length range of 1. 5-4 cm and Gosner developmental stages 25-38. The burst swimming performance of tadpoles at 10 degrees and 24 degrees C was determined by videotaping startle responses with a high-speed video camera at 200 Hz and analysing the sequences frame by frame. Maximum swimming velocity (Umax) and acceleration (Amax) increased with total length (L) at a rate that was proportionally greater than the increase in total length (i.e., positive allometry; exponents >1) and was described by the allometric equations Umax=0.061L1.34 and Amax=1.15L1.11 at 10 degrees C and Umax=0.114L1.34 and Amax=1.54L1. 11 at 24 degrees C. Stride length increased with a total-length exponent of approximately 1 but was unaffected by temperature. Tail-beat frequency was not affected by total length and increased from 7.8+/-0.2 Hz at 10 degrees C to 21.7+/-0.7 Hz at 24 degrees C. Developmental stage did not significantly influence the relationship between total length and Umax or Amax. Furthermore, temperature and the associated changes in water viscosity did not affect the relationship between total length and burst swimming performance. At their Umax, Reynolds numbers ranged from approximately 1,500 in the smaller tadpoles up to 50,000 for the larger animals at 24 degrees C. We suggest the positive allometry of Umax in larval L. peronii was due in part to the increases in tail width (TW) with total length (TW=-1.36L1.66), possibly reflecting the increasing importance of burst swimming performance to survival during larval development. PMID- 10801393 TI - The influence of haemoglobin on behavioural thermoregulation and oxygen consumption in Daphnia carinata. AB - When placed in a temperature gradient, most ectotherms have a strict thermal preference that is lowered on exposure to hypoxia. Branchiopods, small aquatic crustaceans, are known to synthesise haemoglobin (Hb) when exposed to hypoxia; hypoxia can occur diurnally and seasonally in ponds. The effect of Hb on behavioural thermoregulation in the branchiopod Daphnia carinata following exposure to both normoxia and hypoxia was examined. Control animals raised in normoxia (Po2=150 mmHg, [Hb]=0.026+/-0.007 mg g dry wt-1) and Hb-rich animals raised in hypoxia (Po2=70 mmHg, [Hb]=0.080+/-0.017 mg g dry wt-1) were placed (N=30) in a tube (length=500 mm, diameter=8 mm) filled with pond water. In the absence of a thermal gradient, control and Hb-rich animals in normoxic water were uniformly distributed along the tube. The presence of a thermal gradient (13 degrees -28 degrees C) elicited clustering at a preferred temperature, T approximately 23 degrees C for both groups. Exposure to hypoxic water in a thermal gradient resulted in a behavioural shift: T approximately 16 degrees C for controls and T approximately 19 degrees C for Hb-rich animals. Measurements of oxygen consumption (V&d2;o2) at fixed temperatures revealed that Hb is associated with a metabolic acclimation to hypoxia. PMID- 10801394 TI - Whole-body systemic transcapillary filtration rates, coefficients, and isogravimetric capillary pressures in Bufo marinus and Rana catesbeiana. AB - Whole-body and organ-level transcapillary filtration rates and coefficients are virtually unexamined in ectothermal vertebrates. These filtration rates appear to be greater than in mammals when plasma volume shifts and lymphatic function are analyzed. Gravimetric techniques monitoring whole-body mass changes were used to estimate net systemic filtration in Bufo marinus and Rana catesbeiana while perfusing with low-protein Ringer's and manipulating venous pressure. Capillary pressures were estimated from arterial and venous pressures after measuring the venous to arterial resistance ratio of 0.23. The capillary filtration coefficient (CFC) for the two species was 25.2+/-1.47 mL min-1 kg-1 kPa-1. Isogravimetric capillary pressure (Pci), the pressure at which net fluid is neither filtered nor reabsorbed, was 1.12+/-0.054 kPa and was confirmed by an independent method. None of these variables showed a significant interspecific difference. The anuran CFC and Pci are significantly higher than those found using the same method on rats (7.6+/-2.04 mL min-1 kg-1 kPa-1 and 0.3+/-0.37 kPa, respectively) and those commonly reported in mammals. Despite the high CFC, the high Pci predicts that little net filtration will occur at resting in vivo capillary pressures. PMID- 10801395 TI - Ontogeny of osmoregulation in the crayfish Astacus leptodactylus. AB - Osmoregulation was studied during the postembryonic development of Astacus leptodactylus Eschscholtz 1823 in juvenile stages 1-8 and in adults. Juveniles hatch and later stages develop in freshwater or in moderately saline waters. The time of acclimation from freshwater to a saline medium increased from early juveniles to adults. At all stages, it was longer than in comparable stages of marine crustaceans, reflecting the high impermeability of the teguments to water and ions. All stages were able to hyperisoosmoregulate. In freshwater, the ability to hyperosmoregulate was established at hatching and increased during development. The hemolymph osmolality increased from 286 mosm kg-1 in stage 1 juveniles to 419 mosm kg-1 in adults. All stages also hyperregulated at low salinities (7 per thousand and 13 per thousand salinity) and were osmoconformers at higher salinities up to 21 per thousand salinity. The lowest isosmotic salinity tended to increase with the developmental stages. The ability to osmoregulate at hatch and throughout postembryonic development is probably a key physiological adaptation in this and other freshwater crayfish. PMID- 10801396 TI - Physiological effects of a fish oil supplement on captive juvenile tuatara (Sphenodon punctatus). AB - Tuatara (Sphenodon, Order Sphenodontia) are rare New Zealand reptiles whose conservation involves captive breeding. Wild tuatara eat seabirds, which contain high levels of the long-chain n-3 polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). These fatty acids are absent from the captive diet, and consequently, plasma fatty acid composition of wild and captive tuatara differs. This study investigated the effects of incorporating EPA and DHA into the diet of captive juvenile tuatara (Sphenodon punctatus) in an attempt to replicate the plasma fatty acid composition of wild tuatara. Tuatara receiving a fish oil supplement containing EPA and DHA showed overall changes in their plasma fatty acid composition. Phospholipid EPA and DHA increased markedly, reaching 10.0% and 5.9 mol%, respectively, by 18 mo (cf. Ala mutant, which is inactive in GTP hydrolysis. This is the first structure of an SRP component from an archaeon and allows for a detailed comparison with related structures from Escherichia coli and thermophilic bacteria. In particular, differences in the conserved consensus regions for nucleotide binding and the subdomain interfaces are observed, which provide information about the regulation of the GTPase. These interactions allow us to propose a common signalling mechanism for the SRP-SR system. CONCLUSIONS: The overall structure of SRP-GTPases is well conserved between bacteria and archaea, which indicates strong similarities in the regulation of the SRP-targeting pathway. Surprisingly, structure comparisons identified a homodimeric ATP-binding protein as the closest relative. A heterodimer model for the SRP-SR interaction is presented. PMID- 10801495 TI - Enzymes of vancomycin resistance: the structure of D-alanine-D-lactate ligase of naturally resistant Leuconostoc mesenteroides. AB - BACKGROUND: The bacterial cell wall and the enzymes that synthesize it are targets of glycopeptide antibiotics (vancomycins and teicoplanins) and beta lactams (penicillins and cephalosporins). Biosynthesis of cell wall peptidoglycan requires a crosslinking of peptidyl moieties on adjacent glycan strands. The D alanine-D-alanine transpeptidase, which catalyzes this crosslinking, is the target of beta-lactam antibiotics. Glycopeptides, in contrast, do not inhibit an enzyme, but bind directly to D-alanine-D-alanine and prevent subsequent crosslinking by the transpeptidase. Clinical resistance to vancomycin in enterococcal pathogens has been traced to altered ligases producing D-alanine-D lactate rather than D-alanine-D-alanine. RESULTS: The structure of a D-alanine-D lactate ligase has been determined by multiple anomalous dispersion (MAD) phasing to 2.4 A resolution. Co-crystallization of the Leuconostoc mesenteroides LmDdl2 ligase with ATP and a di-D-methylphosphinate produced ADP and a phosphinophosphate analog of the reaction intermediate of cell wall peptidoglycan biosynthesis. Comparison of this D-alanine-D-lactate ligase with the known structure of DdlB D-alanine-D-alanine ligase, a wild-type enzyme that does not provide vancomycin resistance, reveals alterations in the size and hydrophobicity of the site for D-lactate binding (subsite 2). A decrease was noted in the ability of the ligase to hydrogen bond a substrate molecule entering subsite 2. CONCLUSIONS: Structural differences at subsite 2 of the D-alanine-D-lactate ligase help explain a substrate specificity shift (D-alanine to D-lactate) leading to remodeled cell wall peptidoglycan and vancomycin resistance in Gram positive pathogens. PMID- 10801497 TI - Crystal structure of the ffh and EF-G binding sites in the conserved domain IV of Escherichia coli 4.5S RNA. AB - BACKGROUND: Bacterial signal recognition particle (SRP), consisting of 4.5S RNA and Ffh protein, plays an essential role in targeting signal-peptide-containing proteins to the secretory apparatus in the cell membrane. The 4.5S RNA increases the affinity of Ffh for signal peptides and is essential for the interaction between SRP and its receptor, protein FtsY. The 4.5S RNA also interacts with elongation factor G (EF-G) in the ribosome and this interaction is required for efficient translation. RESULTS: We have determined by multiple anomalous dispersion (MAD) with Lu(3+) the 2.7 A crystal structure of a 4.5S RNA fragment containing binding sites for both Ffh and EF-G. This fragment consists of three helices connected by a symmetric and an asymmetric internal loop. In contrast to NMR-derived structures reported previously, the symmetric loop is entirely constituted by non-canonical base pairs. These pairs continuously stack and project unusual sets of hydrogen-bond donors and acceptors into the shallow minor groove. The structure can therefore be regarded as two double helical rods hinged by the asymmetric loop that protrudes from one strand. CONCLUSIONS: Based on our crystal structure and results of chemical protection experiments reported previously, we predicted that Ffh binds to the minor groove of the symmetric loop. An identical decanucleotide sequence is found in the EF-G binding sites of both 4.5S RNA and 23S rRNA. The decanucleotide structure in the 4.5S RNA and the ribosomal protein L11-RNA complex crystals suggests how 4.5S RNA and 23S rRNA might interact with EF-G and function in translating ribosomes. PMID- 10801498 TI - A potential target enzyme for trypanocidal drugs revealed by the crystal structure of NAD-dependent glycerol-3-phosphate dehydrogenase from Leishmania mexicana. AB - BACKGROUND: NAD-dependent glycerol-3-phosphate dehydrogenase (GPDH) catalyzes the interconversion of dihydroxyacetone phosphate and L-glycerol-3-phosphate. Although the enzyme has been characterized and cloned from a number of sources, until now no three-dimensional structure has been determined for this enzyme. Although the utility of this enzyme as a drug target against Leishmania mexicana is yet to be established, the critical role played by GPDH in the long slender bloodstream form of the related kinetoplastid Trypanosoma brucei makes it a viable drug target against sleeping sickness. RESULTS: The 1.75 A crystal structure of apo GPDH from L. mexicana was determined by multiwavelength anomalous diffraction (MAD) techniques, and used to solve the 2.8 A holo structure in complex with NADH. Each 39 kDa subunit of the dimeric enzyme contains a 189-residue N-terminal NAD-binding domain and a 156-residue C-terminal substrate-binding domain. Significant parts of both domains share structural similarity with plant acetohydroxyacid isomeroreductase. The discovery of extra, fatty-acid like, density buried inside the C-terminal domain indicates a possible post-translational modification with an associated biological function. CONCLUSIONS: The crystal structure of GPDH from L. mexicana is the first structure of this enzyme from any source and, in view of the sequence identity of 63%, serves as a valid model for the T. brucei enzyme. The differences between the human and trypanosomal enzymes are extensive, with only 29% sequence identity between the parasite and host enzyme, and support the feasibility of exploiting the NADH-binding site to develop selective inhibitors against trypanosomal GPDH. The structure also offers a plausible explanation for the observed inhibition of the T. brucei enzyme by melarsen oxide, the active form of the trypanocidal drugs melarsoprol and cymelarsan. PMID- 10801500 TI - Present tributes to achievements past PMID- 10801499 TI - Strategies for macromolecular synchrotron crystallography. PMID- 10801748 TI - Circulation online only : may 9, 2000 PMID- 10801749 TI - What we know and don't know about L-arginine and NO. PMID- 10801750 TI - Prevention of coronary restenosis : the evolving evidence base for radiation therapy. PMID- 10801751 TI - Calcineurin is activated in rat hearts with physiological left ventricular hypertrophy induced by voluntary exercise training. AB - BACKGROUND: Calcineurin may play a pivotal role in the signaling of cardiac hypertrophy; since this hypothesis was first put forward, controversial reports have been published using various experimental models. This study was designed to compare the physiological left ventricular hypertrophy (LVH) induced by voluntary exercise with LVH induced by aortic constriction and to determine whether calcineurin participates in the signaling of exercise-induced LVH. METHODS AND RESULTS: Wistar rats were assigned to 1 of the following 5 groups: 10 weeks of voluntary exercise (EX), a sedentary regimen, a 1-week (AC1) or 4-week (AC4) ascending aortic constriction period, or a sham operation. EX rats ran 2.4+/-0.7 km/day voluntarily in specially manufactured cages; this was associated with an increase of LV diastolic dimension and stroke volume. Myocardial calcineurin activity markedly increased in EX rats (46.4+/-8.3 versus 18.4+/-0.5 pmol. min( 1). mg(-1) in sedentary rats; P<0.001) and in AC1 rats (44.9+/-6.7 versus 22.1+/ 3.7 pmol. min(-1). mg(-1) in sham-operated rats; P<0.001), but not in AC4 rats (29.0+/-3.4 pmol. min(-1). mg(-1)). Treatment with cyclosporin A completely inhibited the development of LVH in EX rats, but it only partially attenuated the development of LVH in AC4 rats. CONCLUSIONS: Calcineurin was activated in exercise-induced physiological LVH and in the developing phase of LVH (AC1), but not in decompensated pressure-overload hypertrophy (AC4). Cyclosporin therapy for the prevention of LVH may be harmful because it does not block the development of pathological hypertrophy but rather that of favorable adaptive hypertrophy. PMID- 10801752 TI - ST-Segment recovery adds to the assessment of TIMI 2 and 3 flow in predicting infarct wall motion after thrombolytic therapy. AB - BACKGROUND: Early resolution of ST-segment elevation (ST-segment recovery) is associated with an improved outcome after infarction. Whether this relation is present in patients with Thrombolysis In Myocardial Infarction (TIMI) grade 2 or 3 flow (ie, patent) infarct-related arteries is not known. METHODS AND RESULTS: To examine the associations between time to achieve stable 50% ST-segment recovery assessed by continuous ECG monitoring, infarct artery flow, and infarct zone wall motion (at 48 hours), we studied 134 patients who underwent angiography at 99 (interquartile range 92 to 110) minutes after commencing streptokinase, initiated within 12 hours of onset of symptoms of myocardial infarction. Patients with TIMI 2 or 3 flow who failed to achieve early stable ST-segment recovery (50% ST-segment recovery sustained for > or 4 hours with <100 microV change in the peak lead) by 60 or 90 minutes had a higher fraction of chords in the infarct zone >2 SD below normal wall motion (TIMI 2: 55.5% vs 15.3%, P=0.006; and 56.5% vs 26.8%, P=0.01, respectively; and TIMI 3: 48.8% vs 28.3%, P=0.07; and 51.8% vs 29.9%, P=0.03, respectively). Time to stable ST-segment recovery was a multivariate predictor of infarct zone wall motion (P=0.04) independent of TIMI flow grade and the time from symptom onset to streptokinase therapy. CONCLUSIONS: In patients with TIMI 2 or 3 flow in infarct-related artery, early stable ST segment recovery is associated with improved infarct zone wall motion at 48 hours. ST-segment recovery may provide additional information about the degree of myocyte reperfusion achieved in patients with a patent epicardial infarct-related artery after thrombolytic therapy. PMID- 10801753 TI - Regulation of tissue factor expression in human microvascular endothelial cells by nitric oxide. AB - BACKGROUND: Tissue factor (TF) is a critical determinant of thrombin generation in normal hemostasis and in atherothrombotic disease. Nitric oxide has both antithrombotic and antiatherosclerotic actions in the vasculature, yet its role in the regulation of TF expression has not been examined. METHODS AND RESULTS: To study the effect of endogenous endothelium-derived nitric oxide on TF expression and activity, we induced TF in human microvascular endothelial cells with lipopolysaccharide or interleukin-1beta and observed a dose- and time-dependent increase in TF activity and expression by Northern and Western blotting. L Arginine, the principal substrate for nitric oxide synthases, added to the media suppressed the induction of TF activity significantly (by 66% for lipopolysaccharide induction and by 59% for interleukin-1beta induction) at 24 hours. These changes in activity were accompanied by correlative changes in TF protein and steady-state mRNA. D-Arginine had no effect, and inhibition of endogenous nitric oxide production failed to increase TF expression. CONCLUSIONS: These data suggest that enhanced production of endothelium-derived nitric oxide reduces endotoxin- and cytokine-induced expression of TF and, thereby, the prothrombotic phenotype of the endothelial cell. PMID- 10801754 TI - Elevation of tumor necrosis factor-alpha and increased risk of recurrent coronary events after myocardial infarction. AB - BACKGROUND: Levels of tumor necrosis factor-alpha (TNF-alpha) increase with acute ischemia. However, whether elevations of TNF-alpha in the stable phase after myocardial ischemia (MI) are associated with increased risk of recurrent coronary events is unknown. METHODS AND RESULTS: A nested case-control design was used to compare TNF-alpha levels obtained an average of 8.9 months after initial MI among 272 participants in the Cholesterol And Recurrent Events (CARE) trial who subsequently developed recurrent nonfatal MI or a fatal cardiovascular event (cases) and from an equal number of age- and sex-matched participants who remained free of these events during follow-up (controls). Overall, TNF-alpha levels were significantly higher among cases than controls (2.84 versus 2.57 pg/mL, P=0.02). The excess risk of recurrent coronary events after MI was predominantly seen among those with the highest levels of TNF-alpha, such that those with levels in excess of 4.17 pg/mL (the 95th percentile of the control distribution) had an approximately 3-fold increase in risk (RR=2.7, 95% CI 1.4 to 5.2, P=0.004). Risk estimates were independent of other risk factors and were similar in subgroup analyses limited to cardiovascular death (RR=2.1) or to recurrent nonfatal MI (RR=3.2). CONCLUSIONS: Plasma concentrations of TNF-alpha are persistently elevated among post-MI patients at increased risk for recurrent coronary events. These data support the hypothesis that a persistent inflammatory instability is present among stable patients at increased vascular risk. Novel therapies designed to attenuate inflammation may thus represent a new direction in the treatment of MI. PMID- 10801755 TI - Beneficial effects of intracoronary adenosine as an adjunct to primary angioplasty in acute myocardial infarction. AB - BACKGROUND: The benefits of vessel recanalization in acute myocardial infarction (AMI) are limited by reperfusion damage. In animal models, adenosine limits reperfusion injury, reducing infarct size and improving ventricular function. The aim of this study was to evaluate the safety and feasibility of adenosine adjunct to primary PTCA in AMI. METHODS AND RESULTS: Fifty-four AMI patients undergoing primary PTCA were randomized to intracoronary adenosine or saline. The 2 groups were similar for age, sex, and infarct location. Adenosine administration was feasible and well tolerated. PTCA was successful in all patients and resulted in TIMI 3 flow in all patients given adenosine and in 19 given saline (P<0.05). The no-reflow phenomenon occurred in 1 adenosine patient and in 7 saline patients (P=0.02). Creatine kinase was lower in the adenosine group, and a Q-wave MI developed in 16 adenosine patients and in 23 saline patients (P=0.04). Sixty-four percent of dyssynergic segments improved in the adenosine group and 36% in the saline group (P=0. 001). Function worsened in 2% of dysynergic segments in the adenosine group and in 20% in the saline group (P=0.0001). Adverse cardiac events occurred in 5 patients in the adenosine group and in 13 patients in the saline group (P=0.03). CONCLUSIONS: Intracoronary adenosine administration is feasible and well tolerated in AMI. Adenosine adjunct to primary PTCA ameliorates flow, prevents the no-reflow phenomenon, improves ventricular function, and is associated with a more favorable clinical course. PMID- 10801756 TI - Oral L-arginine in patients with coronary artery disease on medical management. AB - BACKGROUND: Vascular nitric oxide (NO) bioavailability is reduced in patients with coronary artery disease (CAD). We investigated whether oral L-arginine, the substrate for NO synthesis, improves homeostatic functions of the vascular endothelium in patients maintained on appropriate medical therapy and thus might be useful as adjunctive therapy. METHODS AND RESULTS: Thirty CAD patients (29 men; age, 67+/-8 years) on appropriate medical management were randomly assigned to L-arginine (9 g) or placebo daily for 1 month, with crossover to the alternate therapy after 1 month off therapy, in a double-blind study. Nitrogen oxides in serum (as an index of endothelial NO release), flow-mediated brachial artery dilation (as an index of vascular NO bioactivity), and serum cell adhesion molecules (as an index of NO-regulated markers of inflammation) were measured at the end of each treatment period. L-Arginine significantly increased arginine levels in plasma (130+/-53 versus 70+/-17 micromol/L, P<0.001) compared with placebo. However, there was no effect of L-arginine on nitrogen oxides (19.3+/ 7.9 versus 18. 6+/-6.7 micromol/L, P=0.546), on flow-mediated dilation of the brachial artery (11.9+/-6.3% versus 11.4+/-7.9%, P=0.742), or on the cell adhesion molecules E-selectin (47.8+/-15.2 versus 47.2+/-14.4 ng/mL, P=0.601), intercellular adhesion molecule-1 (250+/-57 versus 249+/-57 ng/mL, P=0.862), and vascular cell adhesion molecule-1 (567+/-124 versus 574+/-135 ng/mL, P=0.473). CONCLUSIONS: Oral L-arginine therapy does not improve NO bioavailability in CAD patients on appropriate medical management and thus may not benefit this group of patients. PMID- 10801757 TI - Intracoronary gamma-radiation therapy after angioplasty inhibits recurrence in patients with in-stent restenosis. AB - BACKGROUND: Treatment of in-stent restenosis presents a critical limitation of intracoronary stent implantation. Ionizing radiation has been shown to decrease neointimal formation within stents in animal models and in initial clinical trials. We studied the effects of intracoronary gamma-radiation therapy versus placebo on the clinical and angiographic outcomes of patients with in-stent restenosis. METHODS AND RESULTS: One hundred thirty patients with in-stent restenosis underwent successful coronary intervention and were then blindly randomized to receive either intracoronary gamma-radiation with (192)Ir (15 Gy) or placebo. Four independent core laboratories blinded to the treatment protocol analyzed the angiographic and intravascular ultrasound end points of restenosis. Procedural success and in-hospital and 30-day complications were similar among the groups. At 6 months, patients assigned to radiation therapy required less target lesion revascularization and target vessel revascularization (9 [13.8%] and 17 [26.2%], respectively) compared with patients assigned to placebo (41 [63.1%, P=0.0001] and 44 [67.7%, P=0.0001], respectively). Binary angiographic restenosis was lower in the irradiated group (19% versus 58% for placebo, P=0.001). Freedom from major cardiac events was lower in the radiation group (29.2% versus 67.7% for placebo, P<0.001). CONCLUSIONS: Intracoronary gamma radiation used as adjunct therapy for patients with in-stent restenosis significantly reduces both angiographic and clinical restenosis. PMID- 10801758 TI - Randomized trial of contrast media utilization in high-risk PTCA: the COURT trial. AB - BACKGROUND: Previous in vitro and in vivo studies have suggested an association between thrombus-related events and type of contrast media. Low osmolar contrast agents appear to improve the safety of diagnostic and coronary artery interventional procedures. However, no data are available on PTCA outcomes with an isosmolar contrast agent. METHODS AND RESULTS: A multicenter prospective randomized double-blind trial was performed in 856 high-risk patients undergoing coronary artery intervention. The objective was to compare the isosmolar nonionic dimer iodixanol (n=405) with the low osmolar ionic agent ioxaglate (n=410). A composite variable of in-hospital major adverse clinical events (MACE) was the primary end point. A secondary objective was to evaluate major angiographic and procedural events during and after PTCA. The composite in-hospital primary end point was less frequent in those receiving iodixanol compared with those receiving ioxaglate (5.4% versus 9.5%, respectively; P=0.027). Core laboratory defined angiographic success was more frequent in patients receiving iodixanol (92.2% versus 85. 9% for ioxaglate, P=0.004). There was a trend toward lower total clinical events at 30 days in patients randomized to iodixanol (9.1% versus 13.2% for ioxaglate, P=0.07). Multivariate predictors of in-hospital MACE were use of ioxaglate (P=0.01) and treatment of a de novo lesion (P=0.03). CONCLUSIONS: In this contemporary prospective multicenter trial of PTCA in the setting of acute coronary syndromes, there was a low incidence of in-hospital clinical events for both treatment groups. The cohort receiving the nonionic dimer iodixanol experienced a 45% reduction in in-hospital MACE when compared with the cohort receiving ioxaglate. PMID- 10801759 TI - Right atrial angiographic evaluation of the posterior isthmus: relevance for ablation of typical atrial flutter. AB - BACKGROUND: Gaining anatomic information about the posterior isthmus is not generally part of flutter ablation procedures. We postulated that right atrial (RA) angiography could rationalize the ablation approach by revealing the conformation of the isthmus. METHODS AND RESULTS: In 100 consecutive patients, biplane RA angiography was performed before ablation to guide catheter contact with the isthmus along its length. Angiography showed a wide variation in the width of the isthmus (17 to 54 mm; 31.3+/-7.9), its angle with the inferior vena cava in the right anterior oblique projection (68 degrees to 114 degrees; 90.3+/ 9.0 degrees ), and its lateral position relative to the inferior vena cava in the left anterior oblique projection. A deep sub-Eustachian recess was revealed in 47%, with a mean depth of 4.3+/-2.1 mm (1.5 to 9.4). A Eustachian valve was visualized in 24%. Ablation resulted in bidirectional conduction block (which could be transient) in all, with a median of 2 dragging radiofrequency (RF) applications (2.3+/-2.5 RF applications; 57 degrees C, < or =99 seconds each). Permanent block was achieved in 99%, with a median of 3 RF applications (3.4+/ 3.0). The presence of a Eustachian valve or concave isthmus was associated with statistically more RF applications; the same trend was seen for patients with deep pouches. The number of RF applications decreased statistically throughout the study, indicating a learning curve. No patient had a recurrence after a follow-up of 13+/-11 months. CONCLUSIONS: Right atrial angiography reveals a highly variable isthmus anatomy, often showing particular configurations that can make ablation more laborious. Rational adaptation of the ablation approach to these anatomic findings may contribute to successful ablation. PMID- 10801760 TI - Increased diastolic chamber stiffness during demand ischemia: response to quick length change differentiates rigor-activated from calcium-activated tension. AB - BACKGROUND: Increased diastolic chamber stiffness (increased DCS) during angina (demand ischemia) has been postulated to be generated by increased diastolic myocyte calcium concentration. METHODS AND RESULTS: We reproduced demand ischemia in isolated isovolumically contracting red-cell-perfused rabbit hearts by imposing pacing tachycardia during global low coronary blood flow (32% of baseline). This increased lactate production without increasing oxygen consumption and resulted in increased DCS (isovolumic left ventricular end diastolic pressure [LVEDP] increased 10 mm Hg, P<0. 001, n=38). To determine the mechanism of increased DCS, we assessed responses to a quick-stretch-release maneuver (QSR), in which the intraventricular balloon was rapidly inflated and deflated to achieve a 3% circumferential muscle fiber length change. QSR was first validated as an effective method of discriminating between calcium-driven and rigor-mediated increased DCS. QSR imposed during demand ischemia when DCS had increased (LVEDP pretachycardia versus posttachycardia, 15+/-1 versus 27+/-2 mm Hg, P<0.001, n=6) reduced DCS to pretachycardia values (LVEDP post-QSR, 15+/-1 mm Hg, P<0.001), ie, elicited a response characteristic of rigor, without any component of calcium-generated tension. CONCLUSIONS: A rigor force, possibly resulting from high-energy phosphate depletion and/or an increase in ADP, appears to be the primary mechanism underlying increased DCS in this model of global LV demand ischemia. PMID- 10801761 TI - Adenovirus-based phospholamban antisense expression as a novel approach to improve cardiac contractile dysfunction: comparison of a constitutive viral versus an endothelin-1-responsive cardiac promoter. AB - BACKGROUND: A decrease in sarcoplasmic reticulum Ca(2+) pump (SERCA2) activity is believed to play a role in the impairment of diastolic function of the failing heart. Because the expression ratio of phospholamban (PL) to SERCA2 may be a target to improve contractile dysfunction, a PL antisense RNA strategy was developed under the control of either a constitutive cytomegalovirus (CMV) or an inducible atrial natriuretic factor (ANF) promoter. The latter is upregulated in hypertrophied and failing heart, allowing "induction-by-disease" gene therapy. METHODS AND RESULTS: Part of the PL cDNA was cloned in antisense and sense directions into adenovectors under the control of either a CMV (Ad5CMVPLas and Ad5CMVPLs, respectively) or ANF (Ad5ANFPLas and Ad5ANFPLs, respectively) promoter. Infection of cultured rat neonatal cardiomyocytes with Ad5CMVPLas reduced PL mRNA to 30+/-7% of baseline and PL protein to 24+/-3% within 48 and 72 hours, respectively. The effects were vector dose dependent. Ad5CMVPLas increased the Ca(2+) sensitivity of SERCA2 and reduced the time to 50% recovery of the Ca(2+) transient. A decrease of PL protein was also achieved by infection with Ad5ANFPLas, and the presence of the hypertrophic stimulus, endothelin-1, led to enhanced downregulation of PL. The adenovectors expressing PL sense RNA had no effect on any of the tested parameters. CONCLUSIONS: Vector-mediated PL antisense RNA expression may become a feasible approach to modulate myocyte Ca(2+) homeostasis in the failing heart. The inducible ANF promoter for the first time offers the perspective for induction-by-disease gene therapy, ie, selective expression of therapeutic genes in hypertrophied and failing cardiomyocytes. PMID- 10801762 TI - Gadolinium decreases stretch-induced vulnerability to atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) is frequently associated with atrial dilatation caused by pressure or volume overload. Stretch-activated channels (SACs) have been found in myocardial cells and may promote AF in dilated atria. To prove this hypothesis, we investigated the effect of the SAC blocker gadolinium (Gd(3+)) on AF propensity in the isolated rabbit heart during atrial stretch. METHODS AND RESULTS: In 16 isolated Langendorff-perfused rabbit hearts, the interatrial septum was perforated to equalize biatrial pressures. Caval and pulmonary veins were occluded. Intra-atrial pressure (IAP) was increased in steps of 2 to 3 cm H(2)O by increasing the pulmonary outflow fluid column. Vulnerability to AF was evaluated by 15-second burst pacing at each IAP level. At baseline, IAP needed to be raised to 8.8+/-0.2 cm H(2)O (mean+/-SEM) to induce AF. A dose-dependent decrease in AF vulnerability was observed after Gd(3+) 12.5, 25, and 50 micromol/L was added. AF threshold increased to 19.0+/-0.5 cm H(2)O with Gd(3+) 50 micromol/L (P<0.001 versus baseline). Spontaneous runs of AF occurred in 5 hearts on a rise of IAP to 13.8+/-3.3 cm H(2)O at baseline but never during Gd(3+). Atrial effective refractory period shortened progressively from 78+/-3 ms at 0.5 cm H(2)O to 52+/-3 ms at 20 cm H(2)O (P<0.05). Gd(3+) 50 micromol/L had no significant effect on effective refractory period. CONCLUSIONS: Acute atrial stretch significantly enhances the vulnerability to AF. Gd(3+) reduces the stretch-induced vulnerability to AF in a dose-dependent manner. Block of SAC might represent a novel antiarrhythmic approach to AF under conditions of elevated atrial pressure or volume. PMID- 10801763 TI - Investigation into the sources of superoxide in human blood vessels: angiotensin II increases superoxide production in human internal mammary arteries. AB - BACKGROUND: Increased vascular superoxide anion (.O(2)(-)) production contributes to endothelial dysfunction and hypertension in animal models of cardiovascular disease. Observations in experimental animals suggest that angiotensin II (Ang II) increases.O(2)(-) production by activation of vascular NAD(P)H oxidase. We studied the sources of.O(2)(-) production in human blood vessels and investigated whether, and by what mechanism, Ang II might alter vascular.O(2)(-) production. METHODS AND RESULTS: Internal mammary arteries (IMAs) and saphenous veins (SVs) were collected at the time of cardiac surgery. Vessels were incubated in Krebs buffer at 37 degrees C.O(2)(-) was measured by lucigenin chemiluminescence. Basal. O(2)(-) concentrations were greater in IMAs than SVs. Inhibitors of NAD(P)H oxidase (10 micromol/L to 200 micromol/L diphenyleneiodonium) and xanthine oxidase (1 mmol/L allopurinol) caused reductions in.O(2)(-) concentrations in both IMAs and SVs. Western blotting of superoxide dismutase proteins demonstrated similar expression in IMAs and SVs. Vessels were also incubated in the presence or absence of Ang II (1 pmol/L to 1 micromol/L). Ang II increased.O(2)(-) production in IMAs at 4 hours of incubation (control, 978+/-117 pmol. min(-1). mg(-1); 1 micromol/L of Ang II, 1690+/-213 pmol. min(-1). mg(-1); n=27, P=0.0001, 95% CI 336, 925) but not in SVs. This effect was completely inhibited by coincubation of IMAs with DPI (100 micromol/L), a nonspecific Ang II antagonist ([sar(1), thre(8)]-Ang II, 1 micromol/L) and a specific Ang II type 1 (AT(1)) receptor antagonist (losartan, 1 micromol/L). Conclusions-. O(2)(-) production is greater in human IMAs than in SVs. NAD(P)H oxidase and xanthine oxidase are sources of.O(2)(-) production in these vessels. The vasoactive peptide Ang II increases.O(2)(-) production in human arteries by an AT(1) receptor-dependent mechanism. PMID- 10801764 TI - Clenbuterol-supported dynamic training of skeletal muscle ventricles against systemic load: a key for powerful circulatory assist? AB - BACKGROUND: The profound loss of power that occurs in skeletal muscle after electrical conditioning has been the major limiting factor in its clinical application. This study investigates a 3-fold approach for chronic conditioning of skeletal muscle ventricles (SMVs) combining electrical transformation, dynamic training against systemic load, and pharmacological support with clenbuterol. METHODS AND RESULTS: In 10 adult male goats, SMVs were constructed from latissimus dorsi muscle wrapped around an intrathoracic training device with windkessel characteristics. SMVs were stimulated electrically and trained dynamically by shifting volume against systemic load. Group 1 goats were controls (n=5), and group 2 goats (n=5) were supported with clenbuterol (150 microg 3 times a week). SMV dynamics were recorded weekly over 5 to 8 months: peak pressure (P(max)), stroke volume (SV), volume displacement per minute (VD), stroke work per day (SW/d), and maximum rates of pressure generation, +dP/dt(max), and decay, -dP/dt(max). In group 1, after 149.5+/-2.7 days (n=4), data were P(max)=70.8+/-4.7 mm Hg, SV=3.2+/-1.2 mL, VD=62.3+/-21.1 mL/min, SW/d=0.8+/-0.4 kJ, +dP/dt(max)=64+/-13 mm Hg/s, and -dP/dt(max)=156+/-32 mm Hg/s. These parameters were significantly improved (P<0.007) in the clenbuterol-treated group 2 after 151+/-2.7 days: P(max)=176.2+/-43.8 mm Hg, SV=23.3+/-6.1 mL, VD=568.2+/-186.1 mL/min, SW/d=9.1+/-2.2 kJ, +dP/dt(max)=1134+/-267 mm Hg/s, and dP/dt(max)=1028+/-92 mm Hg/s. In 2 SMVs of group 2, VD increased to 1090 and 1235 mL/min after 202 and 246 days of training, respectively. At termination, myosin heavy chains were totally transformed into myosin heavy chain-1 in all SMVs. CONCLUSIONS: This clenbuterol-supported dynamic training provides powerful SMVs that may have important clinical implications for the treatment of end-stage heart failure by muscular blood pumps. PMID- 10801765 TI - Images in cardiovascular medicine: primary cardiac osteosarcoma in right ventricular outflow tract. PMID- 10801766 TI - Cell transplantation as future therapy for cardiovascular disease?: A workshop of the National Heart, Lung, and Blood Institute. AB - Despite the development of improved therapies and the significant advances in the understanding of the basis of disease pathogenesis, millions of Americans continue to live with life-threatening cardiovascular diseases. Recent breakthroughs suggest exciting directions that are likely to produce more effective therapies for the treatment of cardiovascular disease. One such area, cell transplantation (grafting of healthy cells into the diseased heart), holds enormous potential as an approach to cardiovascular pathophysiology. Once thought to be a scientific long shot, cell transplantation is becoming recognized as a viable strategy to strengthen weak hearts and limit infarct growth. The technology could also be used for the long-term delivery of beneficial recombinant proteins to the heart, which is a strategy to complement molecular biology advances and provide an alternative strategy for gene therapy. On August 24, 1998, the National Heart, Lung, and Blood Institute convened a workshop to discuss the current status of this fast-moving line of research and to explore its promise for treating cardiovascular disease. The participants included basic and clinical researchers, with representatives from academic and commercial research settings. The workshop was designed to establish the state-of-the-art and to equate current research with practical clinical application. The group recommended short- and long-term goals to assist in realizing, in the most expedient manner, the potential utility of cell transplantation for the treatment of cardiovascular disease. A summary of the meeting discussions and recommendations for future areas of research is presented. PMID- 10801767 TI - Stentocarditis. PMID- 10801768 TI - Holt-Oram syndrome vs heart-hand syndrome. PMID- 10801769 TI - Calcium channel blockers activate the interleukin-6 gene via the transcription factors NF-IL6 and NF-kappaB in primary human vascular smooth muscle cells. PMID- 10801770 TI - Possible different involvement of interleukin-1 receptor antagonist gene polymorphism in coronary single vessel disease and myocardial infarction. PMID- 10801771 TI - Is leptin involved in the signaling cascade after myocardial ischemia and reperfusion? PMID- 10801772 TI - Hemostatic function and coronary artery disease. PMID- 10801773 TI - Clopidogrel-associated thrombotic thrombocytopenic purpura identified. PMID- 10801774 TI - Evidence that the beta-amyloid plaques of Alzheimer's disease represent the redox silencing and entombment of abeta by zinc. AB - Abeta binds Zn(2+), Cu(2+), and Fe(3+) in vitro, and these metals are markedly elevated in the neocortex and especially enriched in amyloid plaque deposits of individuals with Alzheimer's disease (AD). Zn(2+) precipitates Abeta in vitro, and Cu(2+) interaction with Abeta promotes its neurotoxicity, correlating with metal reduction and the cell-free generation of H(2)O(2) (Abeta1-42 > Abeta1-40 > ratAbeta1-40). Because Zn(2+) is redox-inert, we studied the possibility that it may play an inhibitory role in H(2)O(2)-mediated Abeta toxicity. In competition to the cytotoxic potentiation caused by coincubation with Cu(2+), Zn(2+) rescued primary cortical and human embryonic kidney 293 cells that were exposed to Abeta1 42, correlating with the effect of Zn(2+) in suppressing Cu(2+)-dependent H(2)O(2) formation from Abeta1-42. Since plaques contain exceptionally high concentrations of Zn(2+), we examined the relationship between oxidation (8-OH guanosine) levels in AD-affected tissue and histological amyloid burden and found a significant negative correlation. These data suggest a protective role for Zn(2+) in AD, where plaques form as the result of a more robust Zn(2+) antioxidant response to the underlying oxidative attack. PMID- 10801775 TI - Activation of MLK2-mediated signaling cascades by polyglutamine-expanded huntingtin. AB - We previously reported that expression of polyglutamine-expanded huntingtin induces apoptosis via c-Jun amino-terminal kinase (JNK) activation in HN33 cells (Liu, Y. F. (1998) J. Biol. Chem. 273, 28873-28822). Extending this study, we now demonstrate a role of mixed-lineage kinase 2 (MLK2), a JNK activator, in polyglutamine-expanded huntingtin-mediated neuronal toxicity. We find that normal huntingtin interacts with MLK2, whereas the polyglutamine expansion interferes with this interaction. Similar to the expression of polyglutamine-expanded huntingtin, expression of MLK2 also induces JNK activation and apoptosis in HN33 cells. Co-expression of dominant negative MLK2 significantly attenuates neuronal apoptosis induced by the mutated huntingtin. Furthermore, over-expression of the N terminus of normal huntingtin partially rescues the neuronal toxicity induced by MLK2. Our results suggest that activation of MLK2-mediated signaling cascades may be partially involved in neuronal death induced by polyglutamine-expanded huntingtin. PMID- 10801776 TI - Requirement of seminolipid in spermatogenesis revealed by UDP-galactose: Ceramide galactosyltransferase-deficient mice. AB - Although seminolipid has long been suspected to play an essential role in spermatogenesis because of its uniquely abundant and temporally regulated expression in the spermatocytes, direct experimental evidence has been lacking. We have tested the hypothesis by examining the testis of the UDP galactose:ceramide galactosyltransferase-deficient mouse, which is incapable of synthesizing seminolipid. Spermatogenesis in homozygous affected males is arrested at the late pachytene stage and the spermatogenic cells degenerate through the apoptotic process. This stage closely follows the phase of rapid seminolipid synthesis in the wild-type mouse. These observations not only provide the first experimental evidence that seminolipid is indeed essential for normal spermatogenesis but also support the broader concept that cell surface glycolipids are important in cellular differentiation and cell-to-cell interaction. PMID- 10801777 TI - Inhibiting amyloid precursor protein C-terminal cleavage promotes an interaction with presenilin 1. AB - Presenilin 1 (PS1) plays a pivotal role in the production of the amyloid-beta protein, which is central to the pathogenesis of Alzheimer's disease. It has been demonstrated that PS1 regulates the gamma-secretase proteolysis of the amyloid precursor protein (APP) C-terminal fragment (APP-C100), which is the final step in amyloid-beta protein production. The mechanism and detailed pathway of this PS1 activity has yet to be fully resolved, but it may be due to a presenilin controlled trafficking of the APP fragment or possibly an inherent PS1 proteolytic activity. We have investigated the possibility of a direct interaction of PS1 and the APP-C100 within the high molecular mass presenilin complex. However, the APP-C100 is rapidly degraded, and if it forms, then any PS1.APP complex is likely to be very transitory. To circumvent this problem, we have utilized the protease inhibitor N-acetyl-leucyl-norleucinal (LLnL) and the lysosomotropic agent NH(4)Cl, which inhibits the turnover of the APP-C100. Under these conditions, levels of the fragment increased appreciably, and as shown by glycerol gradient analysis, the APP-C100 shifted to a higher molecular mass complex that overlapped with PS1. Immunoprecipitation studies demonstrated that a significant population of the APP-C100 co-precipitated with PS1. These findings suggest that PS1 may mediate the shuttling of APP fragments and/or facilitate their presentation for gamma-secretase cleavage through a direct interaction. PMID- 10801778 TI - Overexpression of bamacan/SMC3 causes transformation. AB - Bamacan can occur in certain cell types as either a secreted proteoglycan assembled into basement membranes or as an intracellular protein known as structural maintenance of chromosome 3 (SMC3). To assess the role of this protein in tumorigenesis, we investigated whether induced overexpression of bamacan/SMC3 could transform normal fibroblasts. We generated a full-length cDNA encoding the entire mouse bamacan/SMC3 and demonstrated appropriate transcription and translation into a 146-kDa protein. All the NIH and Balb/c 3T3 murine fibroblasts overexpressing this bamacan/SMC3 transgene generated foci of transformation and acquired anchorage-independent growth. The increased levels of bamacan/SMC3 expression achieved in the transfected fibroblasts were the same as those detected in a series of spontaneously transformed murine and human colon carcinoma cells. Moreover, a 3-4-fold overexpression of bamacan/SMC3 was detected in approximately 70% of human colon carcinoma specimens from matched pairs (n = 19, p < 0.0002) and in a cohort of intestinal tumors from Apc-deficient Min/+ mice. These results support the concept that deregulated expression of bamacan/SMC3 is involved in cell transformation. PMID- 10801779 TI - Deletion mutation in Drosophila ma-l homologous, putative molybdopterin cofactor sulfurase gene is associated with bovine xanthinuria type II. AB - Defective xanthine dehydrogenase (XDH) activity in humans results in xanthinuria and xanthine calculus accumulation in kidneys. Bovine xanthinuria was demonstrated in a local herd and characterized as xanthinuria type II, similar to the Drosophila ma-l mutations, which lose activities of molybdoenzymes, XDH, and aldehyde oxidase, although sulfite oxidase activity is preserved. Linkage analysis located the disease locus at the centromeric region of bovine chromosome 24, where a ma-l homologous, putative molybdopterin cofactor sulfurase gene (MCSU) has been physically mapped. We found that a deletion mutation at tyrosine 257 in MCSU is tightly associated with bovine xanthinuria type II. PMID- 10801780 TI - GI domain-mediated association of the eukaryotic initiation factor 2alpha kinase GCN2 with its activator GCN1 is required for general amino acid control in budding yeast. AB - In response to the starvation of a single amino acid, the budding yeast Saccharomyces cerevisiae activates numerous genes involved in various amino acid biosynthetic pathways, all of which are under the control of transcription factor GCN4. This general amino acid control response is based on de-repressed translation of GCN4 mRNA, which is induced by the activation of the eIF2alpha kinase, GCN2. Although it is known that in vivo activation of GCN2 requires GCN1, the mode of GCN1 action remains to be elucidated at the molecular level. Here, we show that GCN2 interacts with GCN1 via the GI domain, a novel protein-binding module that occurs at the N terminus; mutations to conserved residues of this domain abolish its binding to GCN1. Furthermore, the yeast cells with GCN2 defective in interaction with GCN1 fail to display general control response. A similar phenotype is observed in cells overexpressing the GI domain of GCN2 or its target region on GCN1. Thus, GI domain-mediated association of GCN2 to GCN1 is required for general amino acid control. This finding provides the first insight into the molecular mechanism for the activation of GCN2 by GCN1. PMID- 10801781 TI - Interaction between RNA polymerase and RapA, a bacterial homolog of the SWI/SNF protein family. AB - Recently, we identified a novel Escherichia coli RNA polymerase (RNAP)-associated protein, an ATPase, called RapA (Sukhodolets, M. V. , and Jin, D. J. (1998) J. Biol. Chem. 273, 7018-7023). RapA is a bacterial homolog of SWI2/SNF2. We showed that RapA forms a stable complex with RNAP holoenzyme and that binding to RNAP holoenzyme stimulates the ATPase activity of RapA. We have further analyzed the interactions between purified RapA and the two forms of RNAP: core RNAP and RNAP holoenzyme. We found that RapA interacts with either form of RNAP. However, RapA exhibits higher affinity for core RNAP than for RNAP holoenzyme. Chemical cross linking of the RNAP-RapA complex indicated that the RapA-binding sites are located at the interface between the alpha and beta' subunits of RNAP. Contrary to previously reported results (Muzzin, O., Campbell, E., A., Xia, L., Severinova, E., Darst, S. A., and Severinov, K. (1998) J. Biol. Chem. 273, 15157 15161), our in vivo analysis of a rapA null mutant suggested that RapA is not likely to be directly involved in DNA repair. PMID- 10801782 TI - Roles of tissue transglutaminase in ethanol-induced inhibition of hepatocyte proliferation and alpha 1-adrenergic signal transduction. AB - The mechanisms by which ethanol inhibits hepatocyte proliferation have been a source of some considerable investigation. Our studies have suggested a possible role for tissue transglutaminase (tTG) in this process. Others have shown that tTG has two distinctly different functions: it catalyzes protein cross-linking, which can lead to apoptosis and enhancement of extracellular matrix stability, and it can function as a G protein (Galpha(h)). Under that circumstance, we speculated that the cross-linking activity would be decreased and that it would function to enhance hepatocyte proliferation in response to adrenergic stimulation. Ethanol treatment inhibited hepatocyte proliferation and led to enhanced tTG cross-linking activity, whereas treatment of hepatocytes with an alpha1 adrenergic agonist, phenylephrine, enhanced hepatocyte proliferation while decreasing tTG cross-linking. However, phenylephrine treatment of several hepatoma cell lines had no effect on cellular proliferation or tTG cross-linking activity, and of note, Northern blot analysis demonstrated that whereas primary hepatocytes had high levels of the alpha1beta adrenergic receptor (alpha1BAR) mRNA, the hepatoma cell lines did not have this mRNA. When the Hep G(2) cell line was stably transduced with an expression vector containing the alpha1BR cDNA, the cell line responded to phenylephrine treatment with enhanced proliferation and with decreased tTG cross-linking activity. Ethanol treatment of the alpha1BAR transfected cells suppressed the phospholipase C-mediated signaling pathways, as detected in the phenylephrine-induced Ca(2+) response. These results suggest that phenylephrine stimulation of hepatocyte proliferation appears to be occurring through the alpha1BAR, which is known to be coupled with the tTG G protein moiety, Galpha(h), and that tTG appears to play a significant role in either enhancing or inhibiting hepatocyte proliferation, depending on its cellular location and on whether it functions as a cross-linking enzyme or a G protein. PMID- 10801783 TI - NF-IL6 (C/EBPbeta ) vigorously activates il1b gene expression via a Spi-1 (PU.1) protein-protein tether. AB - Two classes of transcription factors, ETS and bZIP, stand out as key mediators of monocyte commitment and differentiation. The ETS domain factor Spi-1 (also called PU.1) and the bZIP factor NF-IL6 (also called C/EBPbeta) have been shown to be involved in the transcriptional regulation of interleukin-1beta gene (il1b) and other monocyte-specific genes. We now show that these two factors strongly cooperate on the il1b core promoter (-59/+12) in the absence of direct NF-IL6 binding to DNA. Transient transfection assays, using mutated il1b core promoters, showed that the Spi-1, but not the NF-IL6, binding site is absolutely required for functional cooperativity. Furthermore, the NF-IL6 transactivation domain (TAD) is functionally indispensable and more critical than that of Spi-1. Additionally, TAD-deficient NF-IL6 functions as a dominant negative for Spi-1 mediated activation, suggesting the involvement of the bZIP DNA binding domain. This is supported by the demonstration of in vitro interaction between the NF-IL6 bZIP and Spi-1 winged helix-turn-helix (wHTH) DNA binding domains, arguing that NF-IL6 vigorously activates the il1b core promoter via protein-tethered transactivation mediated by Spi-1. PMID- 10801784 TI - Identification of a methyltransferase from Mycobacterium smegmatis involved in glycopeptidolipid synthesis. AB - Glycopeptidolipids (GPLs) are major components of the cell walls of several species of mycobacteria. We have isolated a transposon mutant of Mycobacterium smegmatis that is unable to synthesize mature GPLs and that displays a rough colony morphology. The disrupted gene, mtf1, shares a high degree of homology with several S-adenosylmethionine-dependent methyltransferases. The enzyme encoded by mtf1 is required for the methylation of a single rhamnose residue that forms part of the conserved GPL core structure. This conclusion is supported by the finding that (a) the mutant synthesized only GPLs with undermethylated (either mono- or nonmethylated instead of di- or trimethylated) rhamnose residues; (b) complementation of the mutant with a wild-type copy of mtf1 restored high levels of synthesis of GPLs containing di- and trimethylated rhamnose; and (c) S-adenosylmethionine-dependent methylation of rhamnosylated GPLs could be detected in cell lysates of wild-type cells and mtf1-complemented mutant cells, but not in mutant cells lacking intact mtf1. Structural analysis of wild-type and mutant GPLs suggests that disruption of mtf1 specifically inhibits addition of O-methyl groups to the 3 (or 2)-position of the rhamnose. In the absence of 3-O-methylation, further methylation of GPL rhamnose is apparently inhibited, and overall GPL synthesis is down-regulated by 90%. PMID- 10801785 TI - Examining thrombin hydrolysis of the factor XIII activation peptide segment leads to a proposal for explaining the cardioprotective effects observed with the factor XIII V34L mutation. AB - In the blood coagulation cascade, thrombin cleaves fibrinopeptides A and B from fibrinogen revealing sites for fibrin polymerization that lead to insoluble clot formation. Factor XIII stabilizes this clot by catalyzing the formation of intermolecular cross-links in the fibrin network. Thrombin activates the Factor XIII a(2) dimer by cleaving the Factor XIII activation peptide segment at the Arg(37)-Gly(38) peptide bond. Using a high performance liquid chromatography assay, the kinetic constants K(m), k(cat), and k(cat)/K(m) were determined for thrombin hydrolysis of fibrinogen Aalpha-(7-20), Factor XIII activation peptide (28-41), and Factor XIII activation peptide-(28-41) with a Val(34) to Leu substitution. This Val to Leu mutation has been correlated with protection from myocardial infarction. In the absence of fibrin, the Factor XIII activation peptide-(28-41) exhibits a 10-fold lower k(cat)/K(m) value than fibrinogen Aalpha (7-20). With the Factor XIII V34L mutation, decreases in K(m) and increases in k(cat) produce a 6-fold increase in k(cat)/K(m) relative to the wild-type Factor XIII sequence. A review of the x-ray crystal structures of known substrates and inhibitors of thrombin leads to a hypothesis that the new Leu generates a peptide with more extensive interactions with the surface of thrombin. As a result, the Factor XIII V34L is proposed to be susceptible to wasteful conversion of zymogen to activated enzyme. Premature depletion may provide cardioprotective effects. PMID- 10801786 TI - A role for Asp75 in domain interactions in the Bacillus subtilis response regulator Spo0A. AB - Spo0A is a two-domain response regulator required for sporulation initiation in Bacillus subtilis. Studies on response regulators have focused on the activity of each domain, but very little is known about the mechanism by which the regulatory domain inhibits the activator domain. In this study, we created a single amino acid substitution in the regulatory domain, D75S, which resulted in a dramatic decrease in sporulation in vivo. In vitro studies with the purified Spo0AD75S protein demonstrated that phosphorylation and DNA binding were comparable with wild type Spo0A. However, the mutant was unable to stimulate transcription by final sigma(A)-RNA polymerase from the Spo0A-dependent spoIIG operon promoter. We suggest that the amino acid Asp(75) and/or the region within which it resides, the alpha3-beta4 loop, are involved in the inhibitory interaction between the regulatory and activator domains of Spo0A. PMID- 10801787 TI - mu-Protocadherin, a novel developmentally regulated protocadherin with mucin-like domains. AB - Branching morphogenesis is a central event during the development of kidneys, lungs, and other organs. We previously generated a monoclonal antibody, 3D2-E9, that inhibited branching morphogenesis and caused widespread apoptosis. We now report the purification of its antigen and cloning of its full-length cDNA. Its cDNA encodes an integral membrane protein that contains four cadherin-like ectodomains and a thrice tandemly repeated region enriched in threonine, serine, and proline, similar to those of mucins. We thus term this protein mu protocadherin, reflecting the hybrid nature of its extracellular region. mu Protocadherin is expressed in two forms that are developmentally regulated, with the shorter isoform lacking the mucin-like repeats. Expression of the long isoform in heterologous cells results in adhesion of the expressing cells, suggesting that it is a new cell adhesion molecule. mu-Protocadherin contains both N and O glycosylations. It is expressed at lateral and basal surfaces of epithelia during kidney and lung development and is located in coated pits. Colocalization of mu-protocadherin with beta-catenin was noted primarily at the junction of the lateral and basal membrane. The cytoplasmic domain contains four proline-rich regions, similar to SH3 binding regions. Thus, it is likely that adhesive interactions mediated by mu-protocadherin induce signaling events that regulate branching morphogenesis. PMID- 10801788 TI - Constitutive regulation of cardiac fatty acid metabolism through peroxisome proliferator-activated receptor alpha associated with age-dependent cardiac toxicity. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the nuclear receptor superfamily and mediates the biological effects of peroxisome proliferators. To determine the physiological role of PPARalpha in cardiac fatty acid metabolism, we examined the regulation of expression of cardiac fatty acid-metabolizing proteins using PPARalpha-null mice. The capacity for constitutive myocardial beta-oxidation of the medium and long chain fatty acids, octanoic acid and palmitic acid, was markedly reduced in the PPARalpha null mice as compared with the wild-type mice, indicating that mitochondrial fatty acid catabolism is impaired in the absence of PPARalpha. In contrast, constitutive beta-oxidation of the very long chain fatty acid, lignoceric acid, did not differ between the mice, suggesting that the constitutive expression of enzymes involved in peroxisomal beta-oxidation is independent of PPARalpha(.) Indeed, PPARalpha-null mice had normal levels of the peroxisomal beta-oxidation enzymes except the D-type bifunctional protein. At least seven mitochondrial fatty acid-metabolizing enzymes were expressed at much lower levels in the PPARalpha-null mice, whereas other fatty acid-metabolizing enzymes were present at similar or slightly lower levels in the PPARalpha-null, as compared with wild type mice. Additionally, lower constitutive mRNA expression levels of fatty acid transporters were found in the PPARalpha-null mice, suggesting a role for PPARalpha in fatty acid transport and catabolism. Indeed, in fatty acid metabolism experiments in vivo, myocardial uptake of iodophenyl 9 methylpentadecanoic acid and its conversion to 3-methylnonanoic acid were reduced in the PPARalpha-null mice. Interestingly, a decreased ATP concentration after exposure to stress, abnormal cristae of the mitochondria, abnormal caveolae, and fibrosis were observed only in the myocardium of the PPARalpha-null mice. These cardiac abnormalities appeared to proceed in an age-dependent manner. Taken together, the results presented here indicate that PPARalpha controls constitutive fatty acid oxidation, thus establishing a role for the receptor in cardiac fatty acid homeostasis. Furthermore, altered expression of fatty acid metabolizing proteins seems to lead to myocardial damage and fibrosis, as inflammation and abnormal cell growth control can cause these conditions. PMID- 10801789 TI - Recruitment of a foreign quinone into the A1 site of photosystem I. Altered kinetics of electron transfer in phylloquinone biosynthetic pathway mutants studied by time-resolved optical, EPR, and electrometric techniques. AB - Interruption of the menA or menB gene in Synechocystis sp. PCC 6803 results in the incorporation of a foreign quinone, termed Q, into the A(1) site of photosystem I with a number of experimental indicators identifying Q as plastoquinone-9. A global multiexponential analysis of time-resolved optical spectra in the blue region shows the following three kinetic components: 1) a 3 ms lifetime in the absence of methyl viologen that represents charge recombination between P700(+) and an FeS(-) cluster; 2) a 750-microseconds lifetime that represents electron donation from an FeS(-) cluster to methyl viologen; and 3) an approximately 15-microseconds lifetime that represents an electrochromic shift of a carotenoid pigment. Room temperature direct detection transient EPR studies of forward electron transfer show a spectrum of P700(+) Q( ) during the lifetime of the spin polarization and give no evidence of a significant population of P700(+) FeS(-) for t MKK4 --> JNK kinase cascade during the late stages of mammalian embryogenesis. PMID- 10801799 TI - Activation of NF-kappa B by bradykinin through a Galpha(q)- and Gbeta gamma dependent pathway that involves phosphoinositide 3-kinase and Akt. AB - Recent work has suggested a role for the serine/threonine kinase Akt and IkappaB kinases (IKKs) in nuclear factor (NF)-kappaB activation. In this study, the involvement of these components in NF-kappaB activation through a G protein coupled pathway was examined using transfected HeLa cells that express the B2 type bradykinin (BK) receptor. The function of IKK2, and to a lesser extent, IKK1, was suggested by BK-induced activation of their kinase activities and by the ability of their dominant negative mutants to inhibit BK-induced NF-kappaB activation. BK-induced NF-kappaB activation and IKK2 activity were markedly inhibited by RGS3T, a regulator of G protein signaling that inhibits Galpha(q), and by two Gbetagamma scavengers. Co-expression of Galpha(q) potentiated BK induced NF-kappaB activation, whereas co-expression of either an activated Galpha(q)(Q209L) or Gbeta(1)gamma(2) induced IKK2 activity and NF-kappaB activation without BK stimulation. BK-induced NF-kappaB activation was partially blocked by LY294002 and by a dominant negative mutant of phosphoinositide 3 kinase (PI3K), suggesting that PI3K is a downstream effector of Galpha(q) and Gbeta(1)gamma(2) for NF-kappaB activation. Furthermore, BK could activate the PI3K downstream kinase Akt, whereas a catalytically inactive mutant of Akt inhibited BK-induced NF-kappaB activation. Taken together, these findings suggest that BK utilizes a signaling pathway that involves Galpha(q), Gbeta(1)gamma(2), PI3K, Akt, and IKK for NF-kappaB activation. PMID- 10801800 TI - The roles of sterol regulatory element-binding proteins in the transactivation of the rat ATP citrate-lyase promoter. AB - ATP citrate-lyase (ACL) is a key enzyme supplying acetyl-CoA for fatty acid and cholesterol synthesis. Its expression is drastically up-regulated when an animal is fed a low fat, high carbohydrate diet after prolonged fasting. In this report, we describe the role of sterol regulatory element-binding proteins (SREBPs) in the transactivation of the rat ACL promoter. ACL promoter activity was markedly stimulated by the overexpression of SREBP-1a and, to a lesser extent, by SREBP-2 in Alexander human hepatoma cells. The promoter elements responsive to SREBPs were located within the 55-base pair sequences from -114 to -60. The gel mobility shift assay revealed four SREBP-1a binding sites in this region. Of these four elements, the -102/-94 region, immediately upstream of the inverted Y-box, and the -70/-61 region, just adjacent to Sp1 binding site, played critical roles in SREBPs-mediated stimulation. The mutation in the inverted Y-box and the coexpression of dominant negative nuclear factor-Y (NF-Y) significantly attenuated the transactivation by SREBP-1a, suggesting that NF-Y binding is a prerequisite for SREBPs to activate the ACL promoter. However, the multiple Sp1 binding sites did not affect the transactivation of the ACL promoter by SREBPs. The binding affinity of SREBP-1a to SREs of the ACL promoter also was much higher than that of SREBP-2. The transactivation potencies of the chimeric SREBPs, of which the activation domains (70 amino acids of the amino terminus) were derived from the different species of their carboxyl-terminal region, were similar to those of SREBPs corresponding to their carboxyl termini. Therefore, it is suggested that the carboxyl-terminal portions of SREBPs containing DNA binding domains are important in determining their transactivation potencies to a certain promoter. PMID- 10801801 TI - Ubiquitin-mediated degradation of the proapoptotic active form of bid. A functional consequence on apoptosis induction. AB - Under basal conditions, the proapoptotic protein Bid is a long-lived protein. Pro apoptotic stimuli such as tumor necrosis factor-alpha (TNFalpha) or Fas induce its caspase-8-mediated cleavage into two fragments. The COOH-terminal cleavage fragment of Bid (tBid) becomes localized to mitochondrial membranes and triggers the release of cytochrome c. Here we show that tBid is ubiquitinated and subsequently degraded by the 26 S proteasome. Degradation of tBid is significantly inhibited by the proteasome inhibitors MG-132 and lactacystin. In contrast, caspase-specific or lysosomal inhibitors do not affect tBid stability. Furthermore, mutation of the putative ubiquitin acceptor sites within tBid results in a stabilized protein as assessed by pulse-chase analysis. To address whether tBid degradation might be regulated by interaction with other Bcl-2-like proteins, cotransfection studies were performed. However, neither the presence of proapoptotic Bax nor antiapoptotic Bcl-2 or Bcl-XL affected tBid degradation. Finally, we determined the functional role of tBid degradation. Overexpression of stabilized tBid proteins significantly enhanced cytochrome c release and subsequent apoptosis induction approximately 2-fold compared with wild type tBid. Similarly, tBid-induced apoptosis was considerably amplified by inhibition of tBid degradation using the proteasome-specific inhibitor MG-132. Thus, proteasomal degradation of tBid limits the extent of apoptosis in living cells. PMID- 10801802 TI - Surfactant proteins A and D bind CD14 by different mechanisms. AB - Surfactant proteins A (SP-A) and D (SP-D) are lung collectins that are constituents of the innate immune system of the lung. Recent evidence (Sano, H., Sohma, H., Muta, T., Nomura, S., Voelker, D. R., and Kuroki, Y. (1999) J. Immunol. 163, 387-395) demonstrates that SP-A modulates lipopolysaccharide (LPS) induced cellular responses by direct interaction with CD14. In this report we examined the structural elements of the lung collectins involved in CD14 recognition and the consequences for CD14/LPS interaction. Rat SP-A and SP-D bound CD14 in a concentration-dependent manner. Mannose and EDTA inhibited SP-D binding to CD14 but did not decrease SP-A binding. The SP-A binding to CD14 was completely blocked by a monoclonal antibody that binds to the SP-A neck domain but only partially blocked by an antibody that binds to the SP-A lectin domain. SP-A but not SP-D bound to deglycosylated CD14. SP-D decreased CD14 binding to both smooth and rough LPS, whereas SP-A enhanced CD14 binding to rough LPS and inhibited binding to smooth LPS. SP-A also altered the migration profile of LPS on a sucrose density gradient in the presence of CD14. From these results, we conclude that 1) lung collectins bind CD14, 2) the SP-A neck domain and SP-D lectin domain participate in CD14 binding, 3) SP-A recognizes a peptide component and SP-D recognizes a carbohydrate moiety of CD14, and 4) lung collectins alter LPS/CD14 interactions. PMID- 10801803 TI - Thermal repair of tryptophan synthase mutations in a regulatory intersubunit salt bridge. Evidence from arrhenius plots, absorption spectra, and primary kinetic isotope effects. AB - This work is aimed at understanding how protein structure and conformation regulate activity and allosteric communication in the tryptophan synthase alpha(2)beta(2) complex from Salmonella typhimurium. Previous crystallographic and kinetic results suggest that both monovalent cations and a salt bridge between alpha subunit Asp(56) and beta subunit Lys(167) play allosteric roles. Here we show that mutation of either of these salt bridging residues produced deleterious effects that could be repaired by increased temperature in combination with CsCl or with NaCl plus an alpha subunit ligand, alpha-glycerol 3 phosphate. Arrhenius plots of the activity data under these conditions were nonlinear. The same conditions yielded temperature-dependent changes in the equilibrium distribution of enzyme-substrate intermediates and in primary kinetic isotope effects. We correlate the results with a model in which the mutant enzymes are converted by increased temperature from a low activity, "open" conformation to a high activity, "closed" conformation under certain conditions. The allosteric ligand and different monovalent cations affected the equilibrium between the open and closed forms. The results suggest that alpha subunit Asp(56) and beta subunit Lys(167) are not essential for catalysis and for allosteric communication between the alpha and beta subunits but that their mutual interaction is important in stabilization of the active, closed form of the alpha(2)beta(2) complex. PMID- 10801804 TI - Bacteriorhodpsin experiences light-induced conformational alterations in nonisomerizable C(13)=C(14) pigments. A study with EPR. AB - The mechanism by which bacteriorhodopsin is activated following light absorption is not completely clear. We have detected protein conformational alterations following light absorption by retinal-based chromophores in the bacteriorhodopsin binding site by monitoring the rate of reduction-oxidation reactions of covalently attached spin labels, using EPR spectroscopy. It was found that the reduction reaction with hydroxylamine is light-catalyzed in the A103C-labeled pigment but not in E74C or M163C. The reaction is light-catalyzed even when isomerization of the C(13)=C(14) bond of the retinal chromophore is prevented. The reverse oxidation reaction with molecular oxygen is effective only in apomembrane derived from the mutant A103C. This reaction is light-accelerated following light absorption of the retinal oxime, which occupies the binding site. The light-induced acceleration is evident also in "locked" bacteriorhodopsin in which isomerization around the C(13)=C(14) bond is prevented. It is evident that the chromophore-protein covalent bond is not a prerequisite for protein response. In contrast to the case of the retinal oxime, a reduced C=N bond A103C-labeled pigment did not exhibit acceleration of the oxidation reaction following light absorption. Acceleration was observed, however, following substitution of the polyene by groups that modify the excited state charge delocalization. It is suggested that protein conformational alterations are induced by charge redistribution along the retinal polyene following light absorption. PMID- 10801805 TI - Size-dependent disaggregation of stable protein aggregates by the DnaK chaperone machinery. AB - Classic in vitro studies show that the Hsp70 chaperone system from Escherichia coli (DnaK-DnaJ-GrpE, the DnaK system) can bind to proteins, prevent aggregation, and promote the correct refolding of chaperone-bound polypeptides into native proteins. However, little is known about how the DnaK system handles proteins that have already aggregated. In this study, glucose-6-phosphate dehydrogenase was used as a model system to generate stable populations of protein aggregates comprising controlled ranges of particle sizes. The DnaK system recognized the glucose-6-phosphate dehydrogenase aggregates as authentic substrates and specifically solubilized and refolded the protein into a native enzyme. The efficiency of disaggregation by the DnaK system was high with small aggregates, but the efficiency decreased as the size of the aggregates increased. High folding efficiency was restored by either excess DnaK or substoichiometric amounts of the chaperone ClpB. We suggest a mechanism whereby the DnaK system can readily solubilize small aggregates and refold them into active proteins. With large aggregates, however, the binding sites for the DnaK system had to be dynamically exposed with excess DnaK or the catalytic action of ClpB and ATP. Disaggregation by the DnaK machinery in the cell can solubilize early aggregates that formed accidentally during chaperone-assisted protein folding or that escaped the protection of "holding" chaperones during stress. PMID- 10801806 TI - Arg-274 and Leu-277 of the gamma-aminobutyric acid type A receptor alpha 2 subunit define agonist efficacy and potency. AB - Alanine-scanning mutagenesis and the whole cell voltage clamp technique were used to investigate the function of the extracellular loop between the second and third transmembrane domains (TM2-TM3) of the gamma-aminobutyric acid type A receptor (GABA(A)-R). A conserved arginine residue in the TM2-TM3 loop of the GABA(A)-R alpha(2) subunit was mutated to alanine, and the mutant alpha(2)(R274A) was co-expressed with wild-type beta(1) and gamma(2S) subunits in human embryonic kidney (HEK) 293 cells. The GABA EC(50) was increased by about 27-fold in the mutant receptor relative to receptors containing a wild-type alpha(2) subunit. Similarly, the GABA EC(50) at alpha(2)(L277A)beta(1)gamma(2S) and alpha(2)(K279A)beta(1)gamma(2S) GABA(A)-R combinations was increased by 51- and 4 fold, respectively. The alpha(2)(R274A) or alpha(2)(L277A) mutations also reduced the maximal response of piperidine-4-sulfonic acid relative to GABA by converting piperidine-4-sulfonic acid into a weak partial agonist at the GABA(A)-R. Based on these results, we propose that alpha(2)(Arg-274) and alpha(2)(Leu-277) are crucial to the efficient transduction of agonist binding into channel gating at the GABA(A)-R. PMID- 10801807 TI - Structural requirement of carboxyl-terminal globular domains of laminin alpha 3 chain for promotion of rapid cell adhesion and migration by laminin-5. AB - The basement membrane protein laminin-5, a heterotrimer of laminin alpha3, beta3, and gamma2 chains, potently promotes cellular adhesion and motility. It has been supposed that the carboxyl-terminal globular region of the alpha3 chain consisting of five distinct domains (G1 to G5) is important for its interaction with integrins. To clarify the function of each G domain, we transfected cDNAs for the full-length (wild type (WT)) and five deletion derivatives (DeltaGs) of the alpha3 chain into human fibrosarcoma cell line HT1080, which expressed and secreted the laminin beta3 and gamma2 chains but not the alpha3 chain. The transfectants with the alpha3 chain cDNAs lacking G5 (DeltaG(5)), G4-5 (DeltaG(4 5)), G3-5 (DeltaG(3-5)), and G2-5 (DeltaG(2-5)) secreted laminin-5 variants at levels comparable to that with WT cDNA. However, the transfectant with the cDNA without any G domains (DeltaG(1-5)) secreted little laminin-5, suggesting that the G domains are essential for the efficient assembly and secretion of the heterotrimer alpha3beta3gamma2. The transfectants with WT, DeltaG(5), and DeltaG(4-5) cDNAs survived in serum-free medium longer than those with DeltaG(3 5), DeltaG(2-5), and DeltaG(1-5) cDNAs. The transfectants with WT, DeltaG(5), and DeltaG(4-5) cDNAs secreted apparently the same size of laminin-5, which lacked G4 and G5 due to proteolytic cleavage between G3 and G4, and these laminin-5 forms potently promoted integrin alpha(3)beta(1)-dependent cell adhesion and migration. However, the laminin-5 forms of DeltaG(3-5) and DeltaG(2-5) hardly promoted the cell adhesion and motility. These findings demonstrate that the G3 domain, but not the G4 and G5 domains, of the alpha3 chain is essential for the potent promotion of cell adhesion and motility by laminin-5. PMID- 10801808 TI - Mutation of Ha-Ras C terminus changes effector pathway utilization. AB - In PC12 cells, Ha-Ras modulates multiple effector proteins that induce neuronal differentiation. To regulate these pathways Ha-Ras must be located at the plasma membrane, a process normally requiring attachment of farnesyl and palmitate lipids to the C terminus. Ext61L, a constitutively activated and palmitoylated Ha Ras that lacks a farnesyl group, induced neurites with more actin cytoskeletal changes and lamellipodia than were induced by farnesylated Ha-Ras61L. Ext61L triggered neurite outgrowth was prevented easily by co-expressing inhibitory Rho, Cdc42, or p21-activated kinase but required increased amounts of inhibitory Rac. Compared with Ha-Ras61L, Ext61L caused 2-fold greater Rac GTP binding and phosphatidylinositol 3-kinase activity in membranes, a hyperactivation that explained the numerous lamellipodia and ineffectiveness of Rac(N17). In contrast, Ext61L activated B-Raf kinase and ERK phosphorylation more poorly than Ha-Ras61L. Thus, accentuated differentiation by Ext61L apparently results from heightened activation of one Ras effector (phosphatidylinositol 3-kinase) and suboptimal activation of another (B-Raf). This surprising unbalanced effector activation, without changes in the designated Ras effector domain, indicates the Ext61L C terminal alternations are a new way to influence Ha-Ras-effector utilization and suggest a broader role of the lipidated C terminus in Ha-Ras biological functions. PMID- 10801809 TI - Transcriptional regulation of the yeast PHO8 promoter in comparison to the coregulated PHO5 promoter. AB - Expression of the PHO8 and PHO5 genes that encode a nonspecific alkaline and acid phosphatase, respectively, is regulated in response to the P(i) concentration in the medium by the same transcription factors. Upon induction by phosphate starvation, both promoters undergo characteristic chromatin remodeling, yet the extent of remodeling at the PHO8 promoter is significantly lower than at PHO5. Despite the coordinate regulation of the two promoters, the PHO8 promoter is almost 10 times weaker than PHO5. Here we show that of two Pho4 binding sites that had been previously mapped at the PHO8 promoter in vitro, only the high affinity one, UASp2, is functional in vivo. Activation of the PHO8 promoter is partially Pho2-dependent. However, unlike at PHO5, Pho4 can bind strongly to its binding site in the absence of Pho2 and remodel chromatin in a Pho2-independent manner. Replacement of the inactive UASp1 element by the UASp1 element from the PHO5 promoter results in more extensive chromatin remodeling and a concomitant 2 fold increase in promoter activity. In contrast, replacement of the high affinity UASp2 site with the corresponding site from PHO5 precludes chromatin remodeling completely and as a consequence promoter activation, despite efficient binding of Pho4 to this site. Deletion of the promoter region normally covered by nucleosomes -3 and -2 results in a 2-fold increase in promoter activity, further supporting a repressive role of these nucleosomes. These data show that there can be strong binding of Pho4 to a UAS element without any chromatin remodeling and promoter activation. The close correlation between promoter activity and the extent of chromatin disruption strongly suggests that the low level of PHO8 induction in comparison with PHO5 is partly due to the inability of Pho4 to achieve complete chromatin remodeling at this promoter. PMID- 10801810 TI - Two domains of the epstein-barr virus origin DNA-binding protein, EBNA1, orchestrate sequence-specific DNA binding. AB - The EBNA1 (for Epstein-Barr nuclear antigen 1) protein of Epstein-Barr virus governs the replication and partitioning of the viral genomes during latent infection by binding to specific recognition sites in the viral origin of DNA replication. The crystal structure of the DNA binding portion of the EBNA1 protein revealed that this region comprises two structural motifs; a core domain, which mediates protein dimerization and is structurally homologous to the DNA binding domain of the papillomavirus E2 protein, and a flanking domain, which mediated all the observed sequence-specific contacts. To test the possibility that the EBNA1 core domain plays a role in sequence-specific DNA binding not revealed in the crystal structure, we examined the effects of point mutations in potential hydrogen bond donors located in an alpha-helix of the EBNA1 core domain whose structural homologue in E2 mediates sequence-specific DNA binding. We show that these mutations severely reduce the affinity of EBNA1 for its recognition site, and that the core domain, when expressed in the absence of the flanking domain, has sequence-specific DNA binding activity. Flanking domain residues were also found to contribute to the DNA binding activity of EBNA1. Thus, both the core and flanking domains of EBNA1 play direct roles in DNA recognition. PMID- 10801811 TI - Myeloperoxidase is involved in H2O2-induced apoptosis of HL-60 human leukemia cells. AB - We examined the mechanism of H(2)O(2)-induced cytotoxicity and its relationship to oxidation in human leukemia cells. The HL-60 promyelocytic leukemia cell line was sensitive to H(2)O(2), and at concentrations up to about 20-25 micrometer, the killing was mediated by apoptosis. There was limited evidence of lipid peroxidation, suggesting that the effects of H(2)O(2) do not involve hydroxyl radical. When HL-60 cells were exposed to H(2)O(2) in the presence of the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN), we detected a 12-line electron paramagnetic resonance spectrum assigned to the POBN/POBN(.) N-centered spin adduct previously described in peroxidase-containing cell-free systems. Generation of this radical by HL-60 cells had the same H(2)O(2) concentration dependence as initiation of apoptosis. In contrast, studies with the K562 human erythroleukemia cell line, which is often used for comparison with the HL-60, and with high passaged HL-60 cells (spent HL-60) studied under the same conditions failed to generate POBN(.). Cellular levels of antioxidant enzymes superoxide dismutase, glutathione peroxidase, and catalase did not explain the differences between these cell lines. Interestingly, the K562 and spent HL-60 cells, which did not generate the radical, also failed to undergo H(2)O(2)-induced apoptosis. Based on this we reasoned that the difference in H(2)O(2)-induced apoptosis might be due to the enzyme myeloperoxidase. Only the apoptosis-manifesting HL-60 cells contained appreciable immunoreactive protein or enzymatic activity of this cellular enzyme. When HL-60 cells were incubated with methimazole or 4 aminobenzoic acid hydrazide, which are inhibitors of myeloperoxidase, they no longer underwent H(2)O(2)-induced apoptosis. Hypochlorous acid stimulated apoptosis in both HL-60 and spent HL-60 cells, indicating that another oxidant generated by myeloperoxidase induces apoptosis and that it may be the direct mediator of H(2)O(2)-induced apoptosis. Taken together these observations indicate that H(2)O(2)-induced apoptosis in the HL-60 human leukemia cell is mediated by myeloperoxidase and is linked to a non-Fenton oxidative event marked by POBN(.). PMID- 10801812 TI - Mechanism of inhibition of HIV-1 integrase by G-tetrad-forming oligonucleotides in Vitro. AB - The G-tetrad-forming oligonucleotides and have been identified as potent inhibitors of human immunodeficiency virus type 1 integrase (HIV-1 IN) activity (Rando, R. F., Ojwang, J., Elbaggari, A., Reyes, G. R., Tinder, R., McGrath, M. S., and Hogan, M. E. (1995) J. Biol. Chem. 270, 1754-1760; Mazumder, A., Neamati, N., Ojwang, J. O., Sunder, S., Rando, R. F., and Pommier, Y. (1996) Biochemistry 35, 13762-13771; Jing, N., and Hogan, M. E. (1998) J. Biol. Chem. 273, 34992 34999). To understand the inhibition of HIV-1 IN activity by the G-quartet inhibitors, we have designed the oligonucleotides and, composed of three and four G-quartets with stem lengths of 19 and 24 A, respectively. The fact that increasing the G-quartet stem length from 15 to 24 A kept inhibition of HIV-1 IN activity unchanged suggests that the binding interaction occurs between a GTGT loop domain of the G-quartet inhibitors and a catalytic site of HIV-1 IN, referred to as a face-to-face interaction. Docking the NMR structure of (Jing and Hogan (1998)) into the x-ray structure of the core domain of HIV-1 IN, HIV-1 IN (51-209) (Maignan, S., Guilloteau, J.-P. , Qing, Z.-L., Clement-Mella, C., and Mikol, V. (1998) J. Mol. Biol. 282, 359-368), was performed using the GRAMM program. The statistical distributions of hydrogen bonding between HIV-1 IN and were obtained from the analyses of 1000 random docking structures. The docking results show a high probability of interaction between the GTGT loop residues of the G-quartet inhibitors and the catalytic site of HIV-1 IN, in agreement with the experimental observation. PMID- 10801813 TI - Tandem repeats are involved in G1 domain inhibition of versican expression and secretion and the G3 domain enhances glycosaminoglycan modification and product secretion via the complement-binding protein-like motif. AB - The large aggregating chondroitin sulfate proteoglycans, including aggrecan, versican (PG-M), neurocan, and brevican, are characterized by N-terminal and C terminal globular (or selectin-like) domains known as the G1 and G3 domains, respectively. For this study, we generated a series of expression constructs containing various combinations of chicken versican/PG-M domains and a leading peptide of link protein in order to examine the roles of the G1 and G3 domains in versican function. In transfection studies, we observed that the presence of the G1 domain was sufficient to inhibit product secretion, while the G3 domain enhanced this process. We also demonstrated that the G1 domain inhibited the attachment of glycosaminoglycan chains to the core proteins, while the G3 domain enhanced this process. Further studies revealed that inhibition of secretion by G1 was mediated by its two tandem repeats, while G3's promotion of glycosaminoglycan chain attachment was apparently dependent on G3's complement binding protein (CBP)-like motif. The modulatory effects of these two molecular domains may contribute to versican's biological activities. PMID- 10801814 TI - MAPK upstream kinase (MUK)-binding inhibitory protein, a negative regulator of MUK/dual leucine zipper-bearing kinase/leucine zipper protein kinase. AB - Mitogen-activated protein kinase upstream kinase/dual leucine zipper-bearing kinase/leucine-zipper protein kinase (MUK/DLK/ZPK) is a MAPKKK class protein kinase that induces JNK/SAPK activation. We report here a protein named MBIP that binds to MUK/DLK/ZPK. MUK-binding inhibitory protein (MBIP) contains two tandemly orientated leucine-zipper-like motifs with a cluster of basic amino acids located between the two motifs. MBIP interacts with one of the two leucine-zipper-like motifs of MUK/DLK/ZPK and inhibits the activity of MUK/DLK/ZPK to induce JNK/SAPK activation. Notably, no similar effect was observed with another JNK/SAPK inducing MAPKKK, COT/Tpl-2, showing the specificity of MBIP action. Furthermore, the overexpression of MBIP partially inhibits the activation of JNK by 0.3 m sorbitol in 293T cells. Taken together, these observations indicate that MBIP can function as a regulator of MUK/DLK/ZPK, a finding that may provide a clue to understanding the molecular mechanism of JNK/SAPK activation by hyperosmotic stress. PMID- 10801815 TI - A functionally conserved N-terminal domain of the friend of GATA-2 (FOG-2) protein represses GATA4-dependent transcription. AB - GATA4 is a transcriptional activator of cardiac-restricted promoters and is required for normal cardiac morphogenesis. Friend of GATA-2 (FOG-2) is a multizinc finger protein that associates with GATA4 and represses GATA4-dependent transcription. To better understand the transcriptional repressor activity of FOG 2 we performed a functional analysis of the FOG-2 protein. The results demonstrated that 1) zinc fingers 1 and 6 of FOG-2 are each capable of interacting with evolutionarily conserved motifs within the N-terminal zinc finger of mammalian GATA proteins, 2) a nuclear localization signal (RKRRK) (amino acids 736-740) is required to program nuclear targeting of FOG-2, and 3) FOG-2 can interact with the transcriptional co-repressor, C-terminal-binding protein-2 via a conserved sequence motif in FOG-2 (PIDLS). Surprisingly, however, this interaction with C-terminal-binding protein-2 is not required for FOG-2 mediated repression of GATA4-dependent transcription. Instead, we have identified a novel N-terminal domain of FOG-2 (amino acids 1-247) that is both necessary and sufficient to repress GATA4-dependent transcription. This N-terminal repressor domain is functionally conserved in the related protein, Friend of GATA1. Taken together, these results define a set of evolutionarily conserved mechanisms by which FOG proteins repress GATA-dependent transcription and thereby form the foundation for genetic studies designed to elucidate the role of FOG-2 in cardiac development. PMID- 10801816 TI - The recruitment of Raf-1 to membranes is mediated by direct interaction with phosphatidic acid and is independent of association with Ras. AB - The serine/threonine kinase Raf-1 is an essential component of the MAPK cascade. Activation of Raf-1 by extracellular signals is initiated by association with intracellular membranes. Recruitment of Raf-1 to membranes has been reported to be mediated by direct association with Ras and by the phospholipase D product phosphatidic acid (PA). Here we report that insulin stimulation of HIRcB fibroblasts leads to accumulation of Ras, Raf-1, phosphorylated MEK, phosphorylated MAPK, and PA on endosomal membranes. Mutations that disrupt Raf-PA interactions prevented recruitment of Raf-1 to membranes, whereas disruption of Ras-Raf interactions did not affect agonist-dependent translocation. Expression of a dominant-negative Ras mutant did not prevent insulin-dependent Raf-1 translocation, but inhibited phosphorylation of MAPK. Finally, the PA-binding region of Raf-1 was sufficient to target green fluorescent protein to membranes, and its overexpression blocked recruitment of Raf-1 to membranes and disrupted insulin-dependent MAPK phosphorylation. These results indicate that agonist dependent Raf-1 translocation is primarily mediated by a direct interaction with PA and is independent of association with Ras. PMID- 10801817 TI - Transcriptional regulation of rat scavenger receptor class B type I gene. AB - The scavenger receptor class B type I (SR-BI) mediates the selective transport of lipids from high density lipoprotein to cells and plays an important role in the reverse uptake of cholesterol to the liver and in the delivery of substrates for steroidogenesis in steroidogenic organs. We report here on the isolation and characterization of the upstream promoter region of the rat SR-BI gene. The transcription start site for rat SR-BI was mapped, and DNA sequence analysis revealed the presence of binding sites for the Sp1 family in the proximal 5' flanking region. Analysis of deletion mutants with different 5' lengths revealed that the region between -121 and -90 base pairs from the transcription start site is essential for the efficient transcription of SR-BI. Both Sp1 and Sp3 bind to three GC boxes in the region (-141 to -1 base pairs) in a sequence-specific manner. Mutations in any of the GC boxes decreased efficient transcription from this promoter in MA-10 mouse Leydig tumor cells. The overexpression of Sp1 or Sp3 protein enhanced the rat SR-BI promoter activity. These results indicate that Sp1 family members of transcription factors are essential for transcription of the rat SR-BI gene. PMID- 10801819 TI - A 54-kDa fragment of the Poly(A)-specific ribonuclease is an oligomeric, processive, and cap-interacting Poly(A)-specific 3' exonuclease. AB - We have previously identified a HeLa cell 3' exonuclease specific for degrading poly(A) tails of mRNAs. Here we report on the purification and identification of a calf thymus 54-kDa polypeptide associated with a similar 3' exonuclease activity. The 54-kDa polypeptide was shown to be a fragment of the poly(A) specific ribonuclease 74-kDa polypeptide. The native molecular mass of the nuclease activity was estimated to be 180-220 kDa. Protein/protein cross-linking revealed an oligomeric structure, most likely consisting of three subunits. The purified nuclease activity released 5'-AMP as the reaction product and degraded poly(A) in a highly processive fashion. The activity required monovalent cations and was dependent on divalent metal ions. The RNA substrate requirement was investigated, and it was found that the nuclease was highly poly(A)-specific and that only 3' end-located poly(A) was degraded by the activity. RNA substrates capped with m(7)G(5')ppp(5')G were more efficiently degraded than noncapped RNA substrates. Addition of free m(7)G(5')ppp(5')G cap analogue inhibited poly(A) degradation in vitro, suggesting a functional link between the RNA 5' end cap structure and poly(A) degradation at the 3' end of the RNA. PMID- 10801818 TI - Characterization of the interaction between zyxin and members of the Ena/vasodilator-stimulated phosphoprotein family of proteins. AB - Zyxin contains a proline-rich N-terminal domain that is similar to the C-terminal domain in the ActA protein of the bacteria, Listeria monocytogenes. We screened the entire amino acid sequence of human zyxin for Mena-interacting peptides and found that, as with ActA, proline-rich sequences were the sole zyxin sequences capable of binding to Ena/vasodilator-stimulated phosphoprotein (VASP) family members in vitro. From this information, we tested zyxin mutants in which the proline-rich sequences were altered. The reduction in Mena/VASP binding was confirmed by peptide tests, immunoprecipitation, and ectopic expression of zyxin variants at the surface of mitochondria. By transfection assays we showed that zyxin interaction with Mena/VASP in vivo enhances the production of actin-rich structures at the apical surface of cells. Microinjection into cells of peptides corresponding to the first proline-rich sequence of zyxin caused the loss of Mena/VASP from focal contacts. Furthermore, these peptides reduced the degree of spreading of cells replated after trypsinization. We conclude that zyxin and proteins that harbor similar proline-rich repeats contribute to the positioning of Mena/VASP proteins. The positioning of Ena/VASP family members appears to be important when the actin cytoskeleton is reorganized, such as during spreading. PMID- 10801820 TI - Essential role of dynamin in internalization of M2 muscarinic acetylcholine and angiotensin AT1A receptors. AB - Most G protein-coupled receptors (GPCRs), including the M(1) muscarinic acetylcholine receptor (mAChR), internalize in clathrin-coated vesicles, a process that requires dynamin GTPase. The observation that some GPCRs like the M(2) mAChR and the angiotensin AT(1A) receptor (AT(1A)R) internalize irrespective of expression of dominant-negative K44A dynamin has led to the proposal that internalization of these GPCRs is dynamin-independent. Here, we report that, contrary to what is postulated, internalization of M(2) mAChR and AT(1A)R in HEK 293 cells is dynamin-dependent. Expression of N272 dynamin, which lacks the GTP binding domain, or K535M dynamin, which is not stimulatable by phosphatidylinositol 4, 5-bisphosphate, strongly inhibits internalization of M(1) and M(2) mAChRs and AT(1A)Rs. Expression of kinase-defective K298M c-Src or Y231F,Y597F dynamin (which cannot be phosphorylated by c-Src) reduces M(1) mAChR internalization. Similarly, c-Src inhibitor PP1 as well as the generic tyrosine kinase inhibitor genistein strongly inhibit M(1) mAChR internalization. In contrast, M(2) mAChR internalization is not (or is only slightly) reduced by expression of these constructs or treatment with PP1 or genistein. Thus, dynamin GTPases are not only essential for M(1) mAChR but also for M(2) mAChR and AT(1A)R internalization in HEK-293 cells. Our findings also indicate that dynamin GTPases are differentially regulated by c-Src-mediated tyrosine phosphorylation. PMID- 10801821 TI - Elements of the primary structure of thrombomodulin required for efficient thrombin-activable fibrinolysis inhibitor activation. AB - Deletion and point mutants of soluble thrombomodulin were used to compare and contrast elements of primary structure required for the activation of thrombin activable fibrinolysis inhibitor (TAFI) and protein C. The smallest mutant capable of efficiently promoting TAFI activation contained residues including the c-loop of epidermal growth factor-3 (EGF3) through EGF6. This mutant is 13 residues longer than the smallest mutant that functioned well with protein C; the latter consisted of residues from the interdomain loop connecting EGF3 and EGF4 through EGF6. Alanine point mutants showed no loss of function in protein C activation for mutations within the c-loop of EGF3. In TAFI activation, however, alanine mutations cause a 50% reduction at Tyr-337, 67% reductions at Asp-338 and Leu-339, and 90% or greater reductions at Val-340, Asp-341, and Glu-343. A mutation at Asp-349 in the peptide connecting EGF3 to EGF4 eliminated activity against both TAFI and protein C. Oxidation of Met-388 in the peptide connecting EGF5 to EGF6 reduced the rate of protein C activation by 80% but marginally, if at all, affected the rate of TAFI activation. Mutation at Phe-376 severely reduced protein C activation but only marginally influenced that of TAFI. A Q387P mutation, however, severely reduced both activities. TAFI activation was shown to be Ca(2+)-dependent. The response, unlike that of protein C, was monotonic and was half-maximal at 0.25 mm Ca(2+). Like protein C activation, TAFI activation was eliminated by a monoclonal antibody directed at the thrombin-binding domain (EGF5) but was not affected by one directed at EGF2. Thus, elements of structure in the thrombin-binding domain are needed for the activation of both protein C and TAFI, but more of the primary structure is needed for TAFI activation. In addition, some residues are needed for one of the reactions but not the other. PMID- 10801822 TI - Calcium affinity, cooperativity, and domain interactions of extracellular EF hands present in BM-40. AB - The structure and function of cytosolic Ca(2+)-binding proteins containing EF hands are well understood. Recently, the presence of EF-hands in an extracellular protein was for the first time proven by the structure determination of the EC domain of BM-40 (SPARC (for secreted protein acidic and rich in cysteine)/osteonectin) (Hohenester, E., Maurer, P., Hohenadl, C., Timpl, R., Jansonius, J. N., and Engel, J. (1996) Nat. Struct. Biol. 3, 67-73). The structure revealed a pair of EF-hands with two bound Ca(2+) ions. Two unusual features were noted that distinguish the extracellular EF-hands of BM-40 from their cytosolic counterparts. An insertion of one amino acid into the loop of the first EF-hand causes a variant Ca(2+) coordination, and a disulfide bond connects the helices of the second EF-hand. Here we show that the extracellular EF-hands in the BM-40 EC domain bind Ca(2+) cooperatively and with high affinity. The EC domain is thus in the Ca(2+)-saturated form in the extracellular matrix, and the EF-hands play a structural rather than a regulatory role. Deletion mutants demonstrate a strong interaction between the EC domain and the neighboring FS domain, which contributes about 10 kJ/mol to the free energy of binding and influences cooperativity. This interaction is mainly between the FS domain and the variant EF-hand 1. Certain mutations of Ca(2+)-coordinating residues changed affinity and cooperativity, but others inhibited folding and secretion of the EC domain in a mammalian cell line. This points to a function of EF-hands in extracellular proteins during biosynthesis and processing in the endoplasmic reticulum or Golgi apparatus. PMID- 10801823 TI - S-Nitrosocysteine increases palmitate turnover on Ha-Ras in NIH 3T3 cells. AB - Ha-Ras is modified by isoprenoid on Cys(186) and by reversibly attached palmitates at Cys(181) and Cys(184). Ha-Ras loses 90% of its transforming activity if Cys(181) and Cys(184) are changed to serines, implying that palmitates make important contributions to oncogenicity. However, study of dynamic acylation is hampered by an absence of methods for acutely manipulating Ha-Ras palmitoylation in living cells. S-nitrosocysteine (SNC) and, to a more modest extent, S-nitrosoglutathione were found to rapidly increase [(3)H]palmitate incorporation into cellular or oncogenic Ha-Ras in NIH 3T3 cells. In contrast, SNC decreased [(3)H]palmitate labeling of the transferrin receptor and caveolin. SNC accelerated loss of [(3)H]palmitate from Ha-Ras, implying that SNC stimulated deacylation and permitted subsequent reacylation of Ha-Ras. SNC also decreased Ha-Ras GTP binding and inhibited phosphorylation of the kinases ERK1 and ERK2 in NIH 3T3 cells. Thus, SNC altered two important properties of Ha Ras activation state and lipidation. These results identify SNC as a new tool for manipulating palmitate turnover on Ha-Ras and for studying requirements of repalmitoylation and the relationship between palmitate cycling, membrane localization, and signaling by Ha-Ras. PMID- 10801824 TI - Differential protein phosphorylation in 3T3-L1 adipocytes in response to insulin versus platelet-derived growth factor. No evidence for a phosphatidylinositide 3 kinase-independent pathway in insulin signaling. AB - Insulin regulates glucose metabolism in adipocytes via a phosphatidylinositide 3 kinase (PI3K)-dependent pathway that appears to involve protein phosphorylation. However, the generation of phosphoinositides is not sufficient for insulin action, and it has been suggested that insulin regulation of glucose metabolism may involve both PI3K-dependent and -independent pathways, the latter being insulin specific. To test this hypothesis, we have designed a phosphoprotein screen to study insulin-specific phosphoproteins that may be either downstream or in parallel to PI3K. Nineteen insulin-regulated phosphospots were detected in the cytosol and high speed pellet fractions, only six of which were significantly regulated by platelet-derived growth factor. Importantly, almost all (92%) of the insulin-specific phosphoproteins identified using this approach were sensitive to the PI3K inhibitor wortmannin. Thus, we obtained no evidence for an insulin specific, PI3K-independent signaling pathway. A large proportion (62%) of the insulin-specific phosphoproteins were enriched in the same high speed pellet fraction to which PI3K was recruited in response to insulin. Thus, our data suggest that insulin specifically stimulates the phosphorylation of a novel subset of downstream targets and this may in part be because of the unique localization of PI3K in response to insulin in adipocytes. PMID- 10801826 TI - Platelet-endothelial cell adhesion molecule-1 (CD31), a scaffolding molecule for selected catenin family members whose binding is mediated by different tyrosine and serine/threonine phosphorylation. AB - Platelet-endothelial cell adhesion molecule (PECAM)-1 is a 130-kDa glycoprotein commonly used as an endothelium-specific marker. Evidence to date suggests that PECAM-1 is more than just an endothelial cell marker but is intimately involved in signal transduction pathways. This is mediated in part by phosphorylation of specific tyrosine residues within the ITAM domain of PECAM-1 and by recruitment of adapter and signaling molecules. Recently we demonstrated that PECAM-1/beta catenin association functions to regulate beta-catenin localization and, moreover, to modulate beta-catenin tyrosine phosphorylation levels. Here we show that: 1) not only beta-catenin, but also gamma-catenin is associated with PECAM-1 in vitro and in vivo; 2) PKC enzyme directly phosphorylates purified PECAM-1; 3) PKC-derived PECAM-1 serine/threonine phosphorylation inversely correlates with gamma-catenin association; 4) PECAM-1 recruits gamma-catenin to cell-cell junctions in transfected SW480 cells; and 5) gamma-catenin may recruit PECAM-1 into an insoluble cytoskeletal fraction. These data further support the concept that PECAM-1 functions as a binder and modulator of catenins and provides a molecular mechanism for previously reported PECAM-1/cytoskeleton interactions. PMID- 10801827 TI - Interaction of mitochondrial elongation factor Tu with aminoacyl-tRNA and elongation factor Ts. AB - Elongation factor (EF) Tu promotes the binding of aminoacyl-tRNA (aa-tRNA) to the acceptor site of the ribosome. This process requires the formation of a ternary complex (EF-Tu.GTP.aa-tRNA). EF-Tu is released from the ribosome as an EF-Tu.GDP complex. Exchange of GDP for GTP is carried out through the formation of a complex with EF-Ts (EF-Tu.Ts). Mammalian mitochondrial EF-Tu (EF-Tu(mt)) differs from the corresponding prokaryotic factors in having a much lower affinity for guanine nucleotides. To further understand the EF-Tu(mt) subcycle, the dissociation constants for the release of aa-tRNA from the ternary complex (K(tRNA)) and for the dissociation of the EF-Tu.Ts(mt) complex (K(Ts)) were investigated. The equilibrium dissociation constant for the ternary complex was 18 +/- 4 nm, which is close to that observed in the prokaryotic system. The kinetic dissociation rate constant for the ternary complex was 7.3 x 10(-)(4) s( )(1), which is essentially equivalent to that observed for the ternary complex in Escherichia coli. The binding of EF-Tu(mt) to EF-Ts(mt) is mutually exclusive with the formation of the ternary complex. K(Ts) was determined by quantifying the effects of increasing concentrations of EF-Ts(mt) on the amount of ternary complex formed with EF-Tu(mt). The value obtained for K(Ts) (5.5 +/- 1.3 nm) is comparable to the value of K(tRNA). PMID- 10801828 TI - Nup116p associates with the Nup82p-Nsp1p-Nup159p nucleoporin complex. AB - Nup116p is a GLFG nucleoporin involved in RNA export processes. We show here that Nup116p physically interacts with the Nup82p-Nsp1p-Nup159p nuclear pore subcomplex, which plays a central role in nuclear mRNA export. For this association, a sequence within the C-terminal domain of Nup116p that includes the conserved nucleoporin RNA-binding motif was sufficient and necessary. Consistent with this biochemical interaction, protein A-Nup116p and the protein A-tagged Nup116p C-terminal domain, like the members of the Nup82p complex, localized to the cytoplasmic side of the nuclear pore complex, as revealed by immunogold labeling. Finally, synthetic lethal interactions were found between mutant alleles of NUP116 and all members of the Nup82p complex. Thus, Nup116p consists of three independent functional domains: 1) the C-terminal part interacts with the Nup82p complex; 2) the Gle2p-binding sequence interacts with Gle2p/Rae1p; and 3) the GLFG domain interacts with shuttling transport receptors such as karyopherin-beta family members. PMID- 10801829 TI - Novel interactions between urokinase and its receptor. AB - Urokinase-type plasminogen activator (uPA) binds to its receptor (uPAR) with a K(d) of about 1 nm. The catalytic activity of the complex is apparent at uPA concentrations close to K(d). Other functions of the complex, such as signal transduction, are apparent at much higher concentrations (35-60 nm). In the present study, we show that uPA and recombinant soluble uPAR (suPAR), at concentrations that exceed the K(d) and the theoretical saturation levels (10-80 nm), establish novel interactions that lead to a further increase in the activity of the single-chain uPA (scuPA)/suPAR and two-chain uPA (tcuPA)/suPAR complexes. Experiments performed using dynamic light scattering, gel filtration, and electron microscopy techniques indicate that suPAR forms dimers and oligomers. The three techniques provide evidence that the addition of an equimolar concentration of scuPA leads to the dissociation of these dimers and oligomers. Biacore data show that suPAR dimers and oligomers bind scuPA with decreased affinity when compared with monomers. We postulate that uPAR is present in equilibrium between oligomer/dimer/monomer forms. The binding of uPA to suPAR dimers and oligomers occurs with lower affinity than the binding to monomer. These novel interactions regulate the activity of the resultant complexes and may be involved in uPA/uPAR mediated signal transduction. PMID- 10801830 TI - Inhibition of the thioredoxin-dependent activation of the NADP-malate dehydrogenase and cofactor specificity. AB - The chloroplastic NADP-malate dehydrogenase is activated by reduction of its N- and C-terminal disulfides by reduced thioredoxin. The activation is inhibited by NADP(+), the oxidized form of the cofactor. Previous studies suggested that the C terminal disulfide was involved in this process. Recent structural data pointed toward a possible direct interaction between the C terminus of the oxidized enzyme and the cofactor. In the present study, the relationship between the cofactor specificity for catalysis and for inhibition of activation has been investigated by changing the cofactor specificity of the enzyme by substitution of selected residues of the cofactor-binding site. An NAD-specific thiol regulated MDH was engineered. Its activation was inhibited by NAD(+) but no longer by NADP(+). These results demonstrate that the oxidized cofactor is bound at the same site as the reduced cofactor and support the idea of a direct interaction between the negatively charged C-terminal end of the enzyme and the positively charged nicotinamide ring of the cofactor, in agreement with the structural data. The structural requirements for cofactor specificity are modeled and discussed. PMID- 10801831 TI - Identification by mutagenesis of conserved arginine and tryptophan residues in rat liver carnitine palmitoyltransferase I important for catalytic activity. AB - Carnitine palmitoyltransferase I catalyzes the conversion of long-chain acyl-CoA to acylcarnitines in the presence of l-carnitine. To determine the role of the conserved arginine and tryptophan residues on catalytic activity in the liver isoform of carnitine palmitoyltransferase I (L-CPTI), we separately mutated five conserved arginines and two tryptophans to alanine. Substitution of arginine residues 388, 451, and 606 with alanine resulted in loss of 88, 82, and 93% of L CPTI activity, respectively. Mutants R601A and R655A showed less than 2% of the wild type L-CPTI activity. A change of tryptophan 391 and 452 to alanine resulted in 50 and 93% loss in carnitine palmitoyltransferase activity, respectively. The mutations caused decreases in catalytic efficiency of 80-98%. The residual activity in the mutant L-CPTIs was sensitive to malonyl-CoA inhibition. Mutants R388A, R451A, R606A, W391A, and W452A had no effect on the K(m) values for carnitine or palmitoyl-CoA. However, these mutations decreased the V(max) values for both substrates by 10-40-fold, suggesting that the main effect of the mutations was to decrease the stability of the enzyme-substrate complex. We suggest that conserved arginine and tryptophan residues in L-CPTI contribute to the stabilization of the enzyme-substrate complex by charge neutralization and hydrophobic interactions. The predicted secondary structure of the 100-amino acid residue region of L-CPTI, containing arginines 388 and 451 and tryptophans 391 and 452, consists of four alpha-helices similar to the known three-dimensional structure of the acyl-CoA-binding protein. We predict that this 100-amino acid residue region constitutes the putative palmitoyl-CoA-binding site in L-CPTI. PMID- 10801832 TI - Differentiation of lactotrope precursor GHFT cells in response to fibroblast growth factor-2. AB - The mechanisms that control the emergence of different anterior pituitary cells from a common stem cell population are largely unknown. The immortalized GHFT cells derived from targeted expression of SV40 T antigen to mouse pituitary display characteristics of somatolactotropic progenitors in that they express the transcription factor GHF-1 (Pit-1) but not growth hormone (GH) or prolactin (PRL). We searched for factors capable of inducing lactotropic differentiation of GHFT cells. PRL gene expression was not observed in cells subjected to a variety of stimuli, which induce PRL gene expression in mature lactotropes. Only fibroblast growth factor-2 (FGF-2) was able to initiate the transcription, synthesis, and release of PRL in GHFT cells. However, induction of PRL expression was incomplete in FGF-2-treated cells, suggesting that additional factors are necessary to attain high levels of PRL transcription in fully differentiated lactotropes. We also show that the FGF-2 response element is located in the proximal PRL promoter. Stimulation of PRL expression by FGF-2 requires endogenous Ets factors and these factors as well as GHF-1 are expressed at low levels in the committed precursor, suggesting that these low levels are limiting for full PRL expression. Moreover, FGF-2 effect on lactotrope differentiation is mediated, at least partially, by stimulation of the Ras-signaling pathway. Our results suggest that, indeed, GHFT cells represent a valid model for studying lactotropic differentiation and that FGF-2 could play a key role both in initiating lactotrope differentiation and maintaining PRL expression. PMID- 10801834 TI - The gene for a novel member of the whey acidic protein family encodes three four disulfide core domains and is asynchronously expressed during lactation. AB - Secretion of whey acidic protein (WAP) in milk throughout lactation has previously been reported for a limited number of species, including the mouse, rat, rabbit, camel, and pig. We report here the isolation of WAP from the milk of a marsupial, the tammar wallaby (Macropus eugenii). Tammar WAP (tWAP) was isolated by reverse-phase HPLC and migrates in SDS-polyacrylamide gel electrophoresis at 29.9 kDa. tWAP is the major whey protein, but in contrast to eutherians, secretion is asynchronous and occurs only from approximately days 130 through 240 of lactation. The full-length cDNA codes for a mature protein of 191 amino acids, which is comprised of three four-disulfide core domains, contrasting with the two four-disulfide core domain arrangement in all other known WAPs. A three-dimensional model for tWAP has been constructed and suggests that the three domains have little interaction and could function independently. Analysis of the amino acid sequence suggests the protein belongs to a family of protease inhibitors; however, the predicted active site of these domains is dissimilar to the confirmed active site for known protease inhibitors. This suggests that any putative protease ligand may be unique to either the mammary gland, milk, or gut of the pouch young. Examination of the endocrine regulation of the tWAP gene showed consistently that the gene is prolactin-responsive but that the endocrine requirements for induction and maintenance of tWAP gene expression are different during lactation. PMID- 10801833 TI - A role for protein kinase cepsilon in the inhibitory effect of epidermal growth factor on calcium-stimulated chloride secretion in human colonic epithelial cells. AB - Epidermal growth factor (EGF) inhibits carbachol-induced chloride secretion in T(84) colonic epithelial cells and has been shown to activate phosphatidylinositol (PI) 3-kinase, leading to inhibition of a basolateral potassium conductance. We asked whether the inhibitory effect of EGF on secretion is due to activation of specific isoforms of protein kinase C (PKC) by PI 3 kinase. Western analysis revealed that PKCalpha, gamma, epsilon, eta, mu, lambda/iota, and zeta were expressed in T(84) cells. Ro318220 (an inhibitor active against PKCepsilon, 10 micrometer) but not Go6983 (an inhibitor active against PKCzeta, 10 micrometer) reversed the inhibitory effect of EGF (100 ng/ml) on carbachol-stimulated chloride secretion. EGF induced the rapid translocation of PKCepsilon from the cytoplasm to the membrane. Wortmannin (50 micrometer) and LY294002 (20 nm), which are PI 3-kinase inhibitors that by themselves had no effect on PKCepsilon activity, significantly suppressed PKCepsilon translocation activated by EGF. LY294002 also reversed the inhibitory action of EGF on chloride secretion. PI (3,4)P(2) increased membrane-associated PKCepsilon and reduced carbachol-induced (86)Rb(+) efflux. Antisense oligonucleotides against PKCepsilon decreased PKCepsilon mass and prevented the inhibitory effect of EGF on carbachol induced (86)Rb(+) efflux. Thus, the inhibitory effect of EGF on carbachol-induced chloride secretion involves the activation of PKCepsilon mediated by PI 3-kinase. Our findings contribute to the understanding of the cellular mechanisms that control chloride secretion. PMID- 10801835 TI - The protein kinase C-dependent phosphorylation of serine 166 is controlled by the phospholipid species bound to the phosphatidylinositol transfer protein alpha. AB - The charge isomers of bovine brain PI-TPalpha (i.e. PI-TPalphaI containing a phosphatidylinositol (PI) molecule and PI-TPalphaII containing a phosphatidylcholine (PC) molecule) were phosphorylated in vitro by rat brain protein kinase C (PKC) at different rates. From the double-reciprocal plot, it was estimated that the V(max) values for PI-TPalphaI and II were 2.0 and 6.0 nmol/min, respectively; the K(m) values for both charge isomers were about equal, i.e. 0.7 micrometer. Phosphorylation of charge isomers of recombinant mouse PI TPalpha confirmed that the PC-containing isomer was the better substrate. Phosphoamino acid analysis of in vitro and in vivo (32)P-labeled PI-TPalphas showed that serine was the major site of phosphorylation. Degradation of (32)P labeled PI-TPalpha by cyanogen bromide followed by high pressure liquid chromatography and sequence analysis yielded one (32)P-labeled peptide (amino acids 104-190). This peptide contained Ser-148, Ser-152, and the consensus PKC phosphorylation site Ser-166. Replacement of Ser-166 with an alanine residue confirmed that indeed this residue was the site of phosphorylation. This mutation completely abolished PI and PC transfer activity. This was also observed when Ser 166 was replaced with Asp, implying that this is a key amino acid residue in regulating the function of PI-TPalpha. Stimulation of NIH3T3 fibroblasts by phorbol ester or platelet-derived growth factor induced the rapid relocalization of PI-TPalpha to perinuclear Golgi structures concomitant with a 2-3-fold increase in lysophosphatidylinositol levels. This relocalization was also observed for Myc-tagged wtPI-TPalpha expressed in NIH3T3 cells. In contrast, the distribution of Myc-tagged PI-TPalpha(S166A) and Myc-tagged PI-TPalpha(S166D) were not affected by phorbol ester, suggesting that phosphorylation of Ser-166 was a prerequisite for the relocalization to the Golgi. A model is proposed in which the PKC-dependent phosphorylation of PI-TPalpha is linked to the degradation of PI. PMID- 10801837 TI - Production of recombinant human type I procollagen trimers using a four-gene expression system in the yeast Saccharomyces cerevisiae. AB - The expression of stable recombinant human collagen requires an expression system capable of post-translational modifications and assembly of the procollagen polypeptides. Two genes were expressed in the yeast Saccharomyces cerevisiae to produce both propeptide chains that constitute human type I procollagen. Two additional genes were expressed coding for the subunits of prolyl hydroxylase, an enzyme that post-translationally modifies procollagen and that confers heat (thermal) stability to the triple helical conformation of the collagen molecule. Type I procollagen was produced as a stable heterotrimeric helix similar to type I procollagen produced in tissue culture. A key requirement for glutamate was identified as a medium supplement to obtain high expression levels of type I procollagen as heat-stable heterotrimers in Saccharomyces. Expression of these four genes was sufficient for correct assembly and processing of type I procollagen in a eucaryotic system that does not produce collagen. PMID- 10801836 TI - The oxidative DNA lesion 8,5'-(S)-cyclo-2'-deoxyadenosine is repaired by the nucleotide excision repair pathway and blocks gene expression in mammalian cells. AB - Xeroderma pigmentosum (XP) patients with inherited defects in nucleotide excision repair (NER) are unable to excise from their DNA bulky photoproducts induced by UV radiation and therefore develop accelerated actinic damage, including cancer, on sun-exposed tissue. Some XP patients also develop a characteristic neurodegeneration believed to result from their inability to repair neuronal DNA damaged by endogenous metabolites since the harmful UV radiation in sunlight does not reach neurons. Free radicals, which are abundant in neurons, induce DNA lesions that, if unrepaired, might cause the XP neurodegeneration. Searching for such a lesion, we developed a synthesis for 8,5'-(S)-cyclo-2'-deoxyadenosine (cyclo-dA), a free radical-induced bulky lesion, and incorporated it into DNA to test its repair in mammalian cell extracts and living cells. Using extracts of normal and mutant Chinese hamster ovary (CHO) cells to test for NER and adult rat brain extracts to test for base excision repair, we found that cyclo-dA is repaired by NER and not by base excision repair. We measured host cell reactivation, which reflects a cell's capacity for NER, by transfecting CHO and XP cells with DNA constructs containing a single cyclo-dA or a cyclobutane thymine dimer at a specific site on the transcribed strand of a luciferase reporter gene. We found that, like the cyclobutane thymine dimer, cyclo-dA is a strong block to gene expression in CHO and human cells. Cyclo-dA was repaired extremely poorly in NER-deficient CHO cells and in cells from patients in XP complementation group A with neurodegeneration. Based on these findings, we propose that cyclo-dA is a candidate for an endogenous DNA lesion that might contribute to neurodegeneration in XP. PMID- 10801838 TI - Runt domain factor (Runx)-dependent effects on CCAAT/ enhancer-binding protein delta expression and activity in osteoblasts. AB - Transcription factor CCAAT/enhancer-binding protein delta (C/EBPdelta) is normally associated with acute-phase gene expression. However, it is expressed constitutively in primary osteoblast cultures where it increases insulin-like growth factor I synthesis in a cAMP-dependent way. Here we show that the 3' proximal region of the C/EBPdelta gene promoter contains a binding sequence for Runt domain factor Runx2, which is essential for osteogenesis. This region of the C/EBPdelta promoter directed high reporter gene expression in osteoblasts, and specifically bound Runx2 in osteoblast-derived nuclear extract. C/EBPdelta gene promoter activity was reduced by mutating the Runx binding sequence or by co transfecting with Runx2 antisense expression plasmid, and was enhanced by overexpression of Runx-2. Exposure to prostaglandin E(2) increased Runx-dependent gene transactivation independently of Runx2 binding to DNA. Runx2 bound directly to the carboxyl-terminal region of C/EBPdelta itself, and its ability to drive C/EBPdelta expression was suppressed when C/EBPdelta or its carboxyl-terminal fragment was increased by overexpression. Consistent effects also occurred on C/EBPdelta-dependent increases in gene expression driven by synthetic or insulin like growth factor I gene promoter fragments. These interactions between Runx2 and C/EBPdelta, and their activation by prostaglandin E(2), provide new evidence for their importance during skeletal remodeling, inflammatory bone disease, or fracture repair. PMID- 10801839 TI - Binding of high density lipoprotein (HDL) and discoidal reconstituted HDL to the HDL receptor scavenger receptor class B type I. Effect of lipid association and APOA-I mutations on receptor binding. AB - The binding of apoA-I-containing ligands to the HDL receptor scavenger receptor class B type I (SR-BI) was characterized using two different assays. The first employed conventional binding or competition assays with (125)I-labeled ligands. The second is a new nonradioactive ligand binding assay, in which the receptor associated ligand is detected by quantitative immunoblotting ("immunoreceptor assay"). Using both methods, we observed that the K(d) value for spherical HDL (density = 1.1-1.13 g/ml) was approximately 16 microgram of protein/ml, while the values for discoidal reconstituted HDL (rHDL) containing proapoA-I or plasma apoA I were substantially lower (approximately 0.4-5 microgram of protein/ml). We also observed reduced affinity and/or competition for spherical (125)I-HDL cell association by higher relative to lower density HDL and very poor competition by lipid-free apoA-I and pre-beta-1 HDL. Deletion of either 58 carboxyl-terminal or 59 amino-terminal residues from apoA-I, relative to full-length control apoA-I, resulted in little or no change in the affinity of corresponding rHDL particles. However, rHDL particles containing a double mutant lacking both terminal domains competed poorly with spherical (125)I-HDL for binding to SR-BI. These findings suggest an important role for apoA-I and its conformation/organization within particles in mediating HDL binding to SR-BI and indicate that the NH(2) and COOH termini of apoA-I directly or indirectly contribute independently to binding to SR-BI. PMID- 10801840 TI - Identification of key residues for interaction of vasoactive intestinal peptide with human VPAC1 and VPAC2 receptors and development of a highly selective VPAC1 receptor agonist. Alanine scanning and molecular modeling of the peptide. AB - The widespread neuropeptide vasoactive intestinal peptide (VIP) has two receptors VPAC(1) and VPAC(2). Solid-phase syntheses of VIP analogs in which each amino acid has been changed to alanine (Ala scan) or glycine was achieved and each analog was tested for: (i) three-dimensional structure by ab initio molecular modeling; (ii) ability to inhibit (125)I-VIP binding (K(i)) and to stimulate adenylyl cyclase activity (EC(50)) in membranes from cell clones stably expressing human recombinant VPAC(1) or VPAC(2) receptor. The data show that substituting residues at 14 positions out of 28 in VIP resulted in a >10-fold increase of K(i) or EC(50) at the VPAC(1) receptor. Modeling of the three dimensional structure of native VIP (central alpha-helice from Val(5) to Asn(24) with random coiled N and C terminus) and analogs shows that substitutions of His(1), Val(5), Arg(14), Lys(15), Lys(21), Leu(23), and Ile(26) decreased biological activity without altering the predicted structure, supporting that those residues directly interact with VPAC(1) receptor. The interaction of the analogs with human VPAC(2) receptor is similar to that observed with VPAC(1) receptor, with three remarkable exceptions: substitution of Thr(11) and Asn(28) by alanine increased K(i) for binding to VPAC(2) receptor; substitution of Tyr(22) by alanine increased EC(50) for stimulating adenylyl cyclase activity through interaction with the VPAC(2) receptor. By combining 3 mutations at positions 11, 22, and 28, we developed the [Ala(11,22,28)]VIP analog which constitutes the first highly selective (>1,000-fold) human VPAC(1) receptor agonist derived from VIP ever described. PMID- 10801841 TI - Identification and characterization of human endometase (Matrix metalloproteinase 26) from endometrial tumor. AB - We report the discovery, cloning, and characterization of a novel human matrix metalloproteinase 26 (MMP-26) (matrixin) gene, endometase, an endometrial tumor derived metalloproteinase. Among more than three million expressed sequence tags sequenced, the endometase gene was only obtained from human endometrial tumor cDNA library. Endometase mRNA was expressed specifically in human uterus, not in other tissues/cells tested, e.g. testis, heart, brain, lungs, liver, thymus, and melanoma G361. Endometase protein has a signal peptide, a propeptide domain, and a catalytic domain with a unique "cysteine switch" propeptide sequence, PHCGVPDGSD, and a zinc-binding motif, VATHEIGHSLGLQH. Endometase is 43, 41, 41, and 39% identical to human metalloelastase, stromelysin, collagenase-3, and matrilysin, respectively. The zymogen was expressed and isolated from Escherichia coli as inclusion bodies with a molecular mass of 28 kDa. The identity and homogeneity of the recombinant protein was confirmed by protein N-terminal sequencing, silver stain, and immunoblot analyses. The pro-enzyme was partially activated during the folding process. Endometase selectively cleaved type I gelatin and alpha(1)-proteinase inhibitor; however, it did not digest collagens, laminin, elastin, beta-casein, plasminogen, soybean trypsin inhibitor, or Bowman Birk inhibitor. It hydrolyzed peptide substrates of matrixins and tumor necrosis factor-alpha converting enzyme. Endometase may selectively cleave extracellular matrix proteins, inactivate serpins, and process cytokines. PMID- 10801842 TI - Catalytic peptide of human glutaminyl-tRNA synthetase is essential for its assembly to the aminoacyl-tRNA synthetase complex. AB - Human glutaminyl-tRNA synthetase (QRS) is one of several mammalian aminoacyl-tRNA synthetases (ARSs) that form a macromolecular protein complex. To understand the mechanism of QRS targeting to the multi-ARS complex, we analyzed both exogenous and endogenous QRSs by immunoprecipitation after overexpression of various Myc tagged QRS mutants in human embryonic kidney 293 cells. Whereas a deletion mutant containing only the catalytic domain (QRS-C) was targeted to the multi-ARS complex, a mutant QRS containing only the N-terminal appended domain (QRS-N) was not. Deletion mapping showed that the ATP-binding Rossman fold was necessary for targeting of QRS to the multi-ARS complex. Furthermore, exogenous Myc-tagged QRS C was co-immunoprecipitated with endogenous QRS. Since glutaminylation of tRNA was dramatically increased in cells transfected with the full-length QRS, but not with either QRS-C or QRS-N, both the QRS catalytic domain and the N-terminal appended domain were required for full aminoacylation activity. When QRS-C was overexpressed, arginyl-tRNA synthetase and p43 were released from the multi-ARS complex along with endogenous QRS, suggesting that the N-terminal appendix of QRS is required to keep arginyl-tRNA synthetase and p43 within the complex. Thus, the eukaryote-specific N-terminal appendix of QRS appears to stabilize the association of other components in the multi-ARS complex, whereas the C-terminal catalytic domain is necessary for QRS association with the multi-ARS complex. PMID- 10801843 TI - Smad7 and Smad6 differentially modulate transforming growth factor beta -induced inhibition of embryonic lung morphogenesis. AB - Transforming growth factors beta (TGF-beta) are known negative regulators of lung development, and excessive TGF-beta production has been noted in pulmonary hypoplasia associated with lung fibrosis. Inhibitory Smad7 was recently identified to antagonize TGF-beta family signaling by interfering with the activation of TGF-beta signal-transducing Smad complexes. To investigate whether Smad7 can regulate TGF-beta-induced inhibition of lung morphogenesis, ectopic overexpression of Smad7 was introduced into embryonic mouse lungs in culture using a recombinant adenovirus containing Smad7 cDNA. Although exogenous TGF-beta efficiently reduced epithelial lung branching morphogenesis in control virus infected lung culture, TGF-beta-induced branching inhibition was abolished after epithelial transfer of the Smad7 gene into lungs in culture. Smad7 also prevented TGF-beta-mediated down-regulation of surfactant protein C gene expression, a marker of bronchial epithelial differentiation, in cultured embryonic lungs. Moreover, we found that Smad7 transgene expression blocked Smad2 phosphorylation induced by exogenous TGF-beta ligand in lung culture, indicating that Smad7 exerts its inhibitory effect on both lung growth and epithelial cell differentiation through modulation of TGF-beta pathway-restricted Smad activity. However, the above anti-TGF-beta signal transduction effects were not observed in cultured embryonic lungs with Smad6 adenoviral gene transfer, suggesting that Smad7 and Smad6 differentially regulate TGF-beta signaling in developing lungs. Our data therefore provide direct evidence that Smad7, but not Smad6, prevents TGF-beta-mediated inhibition of both lung branching morphogenesis and cytodifferentiation, establishing the mechanistic basis for Smad7 as a novel target to ameliorate aberrant TGF-beta signaling during lung development, injury, and repair. PMID- 10801844 TI - Prothrombinase acceleration by oxidatively damaged phospholipids. AB - The optimally efficient production of thrombin by the prothrombinase complex relies on suitable positioning of its component factors and substrate on phosphatidylserine-containing lipid membranes. The presence of oxidatively damaged phospholipids in a membrane disrupts the normal architecture of a lipid bilayer and might therefore be expected to interfere with prothrombinase activity. To investigate this possibility, we prepared phosphatidylserine containing lipid vesicles containing oxidized arachidonoyl lipids, and we examined their ability to accelerate thrombin production by prothrombinase. Oxidized arachidonoyl chains caused dose-dependent increases in prothrombinase activity up to 6-fold greater than control values. These increases were completely attenuated by the presence of alpha-tocopherol, gamma-tocopherol, or ascorbate. Over the course of a 300-min oxidation, the ability of arachidonoyl lipids to accelerate prothrombinase peaked at 60 min and then declined to base line levels. These results suggest that instead of being impeded by oxidative membrane damage, prothrombinase activity is enhanced by one or more products of nonenzymatic lipid oxidation. PMID- 10801845 TI - Cell cycle-regulated phosphorylation of the human SIX1 homeodomain protein. AB - Human SIX1 (HSIX1) is a member of the Six class of homeodomain proteins implicated in muscle, eye, head, and brain development. To further understand the role of HSIX1 in the cell cycle and cancer, we developed an HSIX1-specific antibody to study protein expression at various stages of the cell cycle. Our previous work demonstrated that HSIX1 mRNA expression increases as cells exit S phase and that overexpression of HSIX1 can attenuate a DNA damage-induced G(2) cell cycle checkpoint. Overexpression of HSIX1 mRNA was observed in 44% of primary breast cancers and 90% of metastatic lesions. Now we demonstrate that HSIX1 is a nuclear phosphoprotein that becomes hyperphosphorylated at mitosis in both MCF7 cells and in Xenopus extracts. The pattern of phosphorylation observed in mitosis is similar to that seen by treating recombinant HSIX1 with casein kinase II (CK2) in vitro. Apigenin, a selective CK2 inhibitor, diminishes interphase and mitotic phosphorylation of HSIX1. Treatment of MCF7 cells with apigenin leads to a dose-dependent arrest at the G(2)/M boundary, implicating CK2, like HSIX1, in the G(2)/M transition. HSIX1 hyperphosphorylated in vitro by CK2 loses its ability to bind the MEF3 sites of the aldolase A promoter (pM), and decreased binding to pM is observed during mitosis. Because CK2 and HSIX1 have both been implicated in cancer and in cell cycle control, we propose that HSIX1, whose activity is regulated by CK2, is a relevant target of CK2 in G(2)/M checkpoint control and that both molecules participate in the same pathway whose dysregulation leads to cancer. PMID- 10801846 TI - Cardiac phospholipase D2 localizes to sarcolemmal membranes and is inhibited by alpha-actinin in an ADP-ribosylation factor-reversible manner. AB - Myocardial phospholipase D (PLD) has been implicated in the regulation of Ca(2+) mobilization and contractile performance in the heart. However, the molecular identity of this myocardial PLD and the mechanisms that regulate it are not well understood. Using subcellular fractionation and Western blot analysis, we found that PLD2 is the major myocardial PLD and that it localizes primarily to sarcolemmal membranes. A 100-kDa PLD2-interacting cardiac protein was detected using a protein overlay assay employing purified PLD2 and then identified as alpha-actinin using peptide-mass fingerprinting with matrix-assisted laser desorption/ionization mass spectroscopy. The direct association between PLD2 and alpha-actinin was confirmed using an in vitro binding assay and localized to PLD2's N-terminal 185 amino acids. Purified alpha-actinin potently inhibits PLD2 activity (IC(50) = 80 nm) in an interaction-dependent and ADP-ribosylation factor reversible manner. Finally, alpha-actinin co-localizes with actin and with PLD2 in the detergent-insoluble fraction from sarcolemmal membranes. These results suggest that PLD2 is reciprocally regulated in sarcolemmal membranes by alpha actinin and ARF1 and accordingly that a major role for PLD2 in cardiac function may involve reorganization of the actin cytoskeleton. PMID- 10801847 TI - Signal-dependent and -independent degradation of free and NF-kappa B-bound IkappaBalpha. AB - A family of inhibitory IkappaB molecules regulates the activation of the transcription factor NF-kappaB. One member of the IkappaB family, IkappaBalpha, plays a major role in the rapid signal-induced activation of NF-kappaB. IkappaBalpha itself is transcriptionally regulated by NF-kappaB allowing for a tight autoregulatory loop that is both sensitive to and rapidly influenced by NF kappaB activating stimuli. For this pathway to remain primed both for rapid activation of NF-kappaB in the presence of signal and then to suppress NF-kappaB activation once that signal is removed, IkappaBalpha must be exquisitely regulated. The regulation of IkappaBalpha is mainly accomplished through phosphorylation, ubiquitination, and subsequent degradation. The mechanism(s) that regulate IkappaBalpha degradation needs to be able to target IkappaBalpha for degradation in both its NF-kappaB bound and free states in the cell. In this study, we utilize a full-length IkappaBalpha mutant that is unable to associate to RelA/p65. We show that the signal-induced IkappaB kinase (IKK) phosphorylation sites on IkappaBalpha can only significantly influence the regulation of signal dependent but not signal-independent turnover of IkappaBalpha. We also demonstrate that the constitutive carboxyl-terminal casein kinase II phosphorylation sites are necessary for the proper regulation of both signal dependent and -independent turnover of IkappaBalpha. These findings further elucidate how the phosphorylation of IkappaBalpha influences the complex regulatory mechanisms involved in maintaining a sensitive NF-kappaB pathway. PMID- 10801848 TI - The major head protein of bacteriophage T4 binds specifically to elongation factor Tu. AB - The Lit protease in Escherichia coli K-12 strains induces cell death in response to bacteriophage T4 infection by cleaving translation elongation factor (EF) Tu and shutting down translation. Suicide of the cell is timed to the appearance late in the maturation of the phage of a short peptide sequence in the major head protein, the Gol peptide, which activates proteolysis. In the present work we demonstrate that the Gol peptide binds specifically to domains II and III of EF Tu, creating the unique substrate for the Lit protease, which then cleaves domain I, the guanine nucleotide binding domain. The conformation of EF-Tu is important for binding and Lit cleavage, because both are sensitive to the identity of the bound nucleotide, with GDP being preferred over GTP. We propose that association of the T4 coat protein with EF-Tu plays a role in phage head assembly but that this association marks infected cells for suicide when Lit is present. Based on these data and recent observations on human immunodeficiency virus type 1 maturation, we speculate that associations between host translation factors and coat proteins may be integral to viral assembly in both prokaryotes and eukaryotes. PMID- 10801849 TI - Identification and cloning of two histone fold motif-containing subunits of HeLa DNA polymerase epsilon. AB - HeLa DNA polymerase epsilon (pol epsilon), possibly involved in both DNA replication and DNA repair, was previously isolated as a complex of a 261-kDa catalytic subunit and a tightly bound 59-kDa accessory protein. Saccharomyces cerevisiae pol epsilon, however, consists of four subunits: a 256-kDa catalytic subunit with 39% identity to HeLa pol epsilon p261, a 80-kDa subunit (DPB2) with 26% identity to HeLa pol epsilon p59, a 23-kDa subunit (DPB3), and a 22-kDa subunit (DPB4). We report here the identification and the cloning of two additional subunits of HeLa pol epsilon, p17, and p12. Both proteins contain histone fold motifs which are present also in S. cerevisiae DPB4 and DPB3. The histone fold motifs of p17 and DPB4 are related to that of subunit A of the CCAAT binding factor, whereas the histone fold motifs found in p12 and DPB3 are homologous to that in subunit C of CCAAT binding factor. p17 together with p12, but not p17 or p12 alone, interact with both p261 and p59 subunits of HeLa pol epsilon. The genes for p17 and p12 can be assigned to chromosome locations 9q33 and 2p12, respectively. PMID- 10801850 TI - A molecular dissection of caveolin-1 membrane attachment and oligomerization. Two separate regions of the caveolin-1 C-terminal domain mediate membrane binding and oligomer/oligomer interactions in vivo. AB - Caveolins form interlocking networks on the cytoplasmic face of caveolae. The cytoplasmically directed N and C termini of caveolins are separated by a central hydrophobic segment, which is believed to form a hairpin within the membrane. Here, we report that the caveolin scaffolding domain (CSD, residues 82-101), and the C terminus (residues 135-178) of caveolin-1 are each sufficient to anchor green fluorescent protein (GFP) to membranes in vivo. We also show that the first 16 residues of the C terminus (i.e. residues 135-150) are necessary and sufficient to attach GFP to membranes. When fused to the caveolin-1 C terminus, GFP co-localizes with two trans-Golgi markers and is excluded from caveolae. In contrast, the CSD targets GFP to caveolae, albeit less efficiently than full length caveolin-1. Thus, caveolin-1 contains at least two membrane attachment signals: the CSD, dictating caveolar localization, and the C terminus, driving trans-Golgi localization. Additionally, we find that caveolin-1 oligomer/oligomer interactions require the distal third of the caveolin-1 C terminus. Thus, the caveolin-1 C-terminal domain has two separate functions: (i) membrane attachment (proximal third) and (ii) protein/protein interactions (distal third). PMID- 10801851 TI - Ultraviolet light-induced stimulation of the JNK mitogen-activated protein kinase in the absence of src family tyrosine kinase activation. AB - In T cells, the JNK mitogen-activated protein kinase is activated by simultaneous stimulation of the T-cell receptor and CD28 or by a number of stress stimuli including ultraviolet light, hydrogen peroxide, and anisomycin. Lck, a Src family kinase, is essential for T-cell receptor-mediated activation of JNK. We asked whether Lck was also involved in stress-mediated activation of JNK. JNK was activated by ultraviolet light irradiation in all of the four T-cell lines we examined, but Lck was not. Additionally, JNK activation by ultraviolet light, hydrogen peroxide, and anisomycin was completely normal in T cells lacking Lck. These data suggest that Lck is not activated by ultraviolet light irradiation, nor is it required for JNK activation in T cells by any of the stress stimuli we tested. We also examined JNK activation by ultraviolet light in mouse fibroblasts expressing no known Src kinases. The activation of JNK by ultraviolet light was completely normal in these cells. Finally, treatment of lymphoid and epithelial cells with a Src kinase family inhibitor PP2-reduced tyrosine phosphorylation of cellular proteins markedly without affecting ultraviolet light-induced activation of JNK. These results suggest that Src kinases are not essential for ultraviolet light-induced activation of JNK in a diverse variety of cell types. PMID- 10801852 TI - Distinct regions of MAT1 regulate cdk7 kinase and TFIIH transcription activities. AB - The transcription/DNA repair factor TFIIH may be resolved into at least two subcomplexes: the core TFIIH and the cdk-activating kinase (CAK) complex. The CAK complex, which is also found free in the cell, is composed of cdk7, cyclin H, and MAT1. In the present work, we found that the C terminus of MAT1 binds to the cdk7 x cyclin H complex and activates the cdk7 kinase activity. The median portion of MAT1, which contains a coiled-coil motif, allows the binding of CAK to the TFIIH core through interactions with both XPD and XPB helicases. Furthermore, using recombinant TFIIH complexes, it is demonstrated that the N-terminal RING finger domain of MAT1 is crucial for transcription activation and participates to the phosphorylation of the C-terminal domain of the largest subunit of the RNA polymerase II. PMID- 10801853 TI - Human factor XII binding to the glycoprotein Ib-IX-V complex inhibits thrombin induced platelet aggregation. AB - Factor XII deficiency has been postulated to be a risk factor for thrombosis suggesting that factor XII is an antithrombotic protein. The biochemical mechanism leading to this clinical observation is unknown. We have previously reported high molecular weight kininogen (HK) inhibition of thrombin-induced platelet aggregation by binding to the platelet glycoprotein (GP) Ib-IX-V complex. Although factor XII will bind to the intact platelet through GP Ibalpha (glycocalicin) without activation, we now report that factor XIIa (0. 37 microm), but not factor XII zymogen, is required for the inhibition of thrombin-induced platelet aggregation. Factor XIIa had no significant effect on SFLLRN-induced platelet aggregation. Moreover, an antibody to the thrombin site on protease activated receptor-1 failed to block factor XII binding to platelets. Inhibition of thrombin-induced platelet aggregation was demonstrated with factor XIIa but not with factor XII zymogen or factor XIIf, indicating that the conformational exposure of the heavy chain following proteolytic activation is required for inhibition. However, inactivation of the catalytic activity of factor XIIa did not affect the inhibition of thrombin-induced platelet aggregation. Factor XII showed displacement of biotin-labeled HK (30 nm) binding to gel-filtered platelets and, at concentrations of 50 nm, was able to block 50% of the HK binding, suggesting involvement of the GP Ib complex. Antibodies to GP Ib and GP IX, which inhibited HK binding to platelets, did not block factor XII binding. However, using a biosensor, which monitors protein-protein interactions, both HK and factor XII bind to GP Ibalpha. Factor XII may serve to regulate thrombin binding to the GP Ib receptor by co-localizing with HK, to control the extent of platelet aggregation in vivo. PMID- 10801854 TI - GCIP, a novel human grap2 and cyclin D interacting protein, regulates E2F mediated transcriptional activity. AB - Regulation of mammalian cell growth and proliferation is governed through receptor-mediated signaling networks that ultimately converge on the cell cycle machinery. Adaptor proteins play essential roles in the formation of intracellular signaling complexes, relaying extracellular signals from the plasma membrane to the nucleus of a cell. The leukocyte-specific adaptor protein Grap2 is a central linker protein in immune cell signaling and activation. Using Grap2 as bait protein, we identified a novel human protein, GCIP (Grap2 cyclin-D interacting protein). We found that GCIP bound to Grap2 in both yeast two-hybrid assays and in mammalian cells through binding to the COOH-terminal unique domain and SH3 domain (designated QC domain) of Grap2. GCIP also associated with cyclin D both in vitro and in vivo. The expression of GCIP was found in all human tissues examined with the highest level of expression in the heart, muscle, peripheral blood leukocytes, and brain. Furthermore, phosphorylation of retinoblastoma protein by cyclin D-dependent protein kinase was reduced and E2F1 mediated transcription activity was inhibited in cells transfected with GCIP. High level expression of GCIP in terminally differentiated tissues and the inhibition of E2F1 transcription activation suggest that GCIP could play an important role in controlling cell differentiation and proliferation. PMID- 10801855 TI - Phenotypic screening of mutations in Pmr1, the yeast secretory pathway Ca2+/Mn2+ ATPase, reveals residues critical for ion selectivity and transport. AB - Thirty-five mutations were generated in the yeast secretory pathway/Golgi ion pump, Pmr1, targeting oxygen-containing side chains within the predicted transmembrane segments M4, M5, M6, M7, and M8, likely to be involved in coordination of Ca(2+) and Mn(2+) ions. Mutants were expressed in low copy number in a yeast strain devoid of endogenous Ca(2+) pumps and screened for loss of Ca(2+) and Mn(2+) transport on the basis of hypersensitivity to 1, 2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) and Mn(2+) toxicity, respectively. Three classes of mutants were found: mutants indistinguishable from wild type (Class 1), mutants indistinguishable from the pmr1 null strain (Class 2), and mutants with differential sensitivity to BAPTA and Mn(2+) toxicity (Class 3). We show that Class 1 mutants retain normal/near normal properties, including (45)Ca transport, Golgi localization, and polypeptide conformation. In contrast, Class 2 mutants lacked any detectable (45)Ca transport; of these, a subset also showed defects in trafficking and protein folding, indicative of structural problems. Two residues identified as Class 2 mutants in this screen, Asn(774) and Asp(778) in M6, also play critical roles in related ion pumps and are therefore likely to be common architectural components of the cation-binding site. Class 3 mutants appear to have altered selectivity for Ca(2+) and Mn(2+) ions, as exemplified by mutant Q783A in M6. These results demonstrate the utility of phenotypic screening in the identification of residues critical for ion transport and selectivity in cation pumps. PMID- 10801856 TI - Manganese selectivity of pmr1, the yeast secretory pathway ion pump, is defined by residue gln783 in transmembrane segment 6. Residue Asp778 is essential for cation transport. AB - We have solubilized and purified the histidine-tagged yeast secretory pathway/Golgi ion pump Pmr1 to near homogeneity in one step, using nickel affinity chromatography. The purified pump demonstrates both Ca(2+)- and Mn(2+) dependent ATP hydrolysis and phosphoenzyme intermediate formation in forward (ATP) and reverse (P(i)) directions. This preparation has allowed us to examine, in detail, the properties of mutations D778A and Q783A in transmembrane segment M6 of Pmr1. In phenotypic screens of Ca(2+) chelator and Mn(2+) toxicity reported separately (Wei, Y., Chen, J., Rosas, G., Tompkins, D.A., Holt, P.A., and Rao, R. (2000) J. Biol. Chem. 275, XXXX-XXXX), D778A was a loss-of-function mutant apparently defective for transport of both Ca(2+) and Mn(2+), whereas mutant Q783A displayed a differential sensitivity consistent with the selective loss of Mn(2+) transport. We show that mutant D778A is devoid of cation-dependent ATP hydrolytic activity and phosphoenzyme formation from ATP. However, reverse phosphorylation from P(i) is preserved but is insensitive to inhibition by Ca(2+) or Mn(2+) ions, which is evidence for a specific inability to bind cations in this mutant. We also show that Ca(2+) can activate ATP hydrolysis in the purified Q783A mutant, with a half-maximal concentration of 0.06 micrometer, essentially identical to that of wild type (0.07 micrometer). Mn(2+) activation of ATP hydrolysis was half-maximal at 0.02 micrometer in wild type, establishing a normal selectivity profile of Mn(2+) > Ca(2+). Strikingly, Mn(2+)-ATPase in the Q783A mutant was nearly abolished, even at concentrations of up to 10 micrometer. These results were confirmed in assays of phosphoenzyme intermediates. Molecular modeling of the packing between helices M4 and M6 suggests that residue Gln(783) in M6 may form a critical hydrophobic interaction with Val(335) in M4, such that the Ala substitution modifies the packing or tilt of the helices and thus the ion pore. The data emphasize the critical role of transmembrane segment M6 in defining the cation binding pocket of P-type ATPases. PMID- 10801857 TI - Is a closing "GA pair" a rule for stable loop-loop RNA complexes? AB - RNA hairpin aptamers specific for the trans-activation-responsive (TAR) RNA element of human immunodeficiency virus type 1 were identified by in vitro selection (Duconge, F., and Toulme, J. J. (1999) RNA 5, 1605-1614). The high affinity sequences selected at physiological magnesium concentration (3 mm) were shown to form a loop-loop complex with the targeted TAR RNA. The stability of this complex depends on the aptamer loop closing "GA pair" as characterized by preliminary electrophoretic mobility shift assays. Thermal denaturation monitored by UV-absorption spectroscopy and binding kinetics determined by surface plasmon resonance show that the GA pair is crucial for the formation of the TAR-RNA aptamer complex. Both thermal denaturation and surface plasmon resonance experiments show that any other "pairs" leads to complexes whose stability decreases in the order AG > GG > GU > AA > GC > UA >> CA, CU. The binding kinetics indicate that stability is controlled by the off-rate rather than by the on-rate. Comparison with the complex formed with the TAR* hairpin, a rationally designed TAR RNA ligand (Chang, K. Y., and Tinoco, I. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 8705-8709), demonstrates that the GA pair is a key determinant which accounts for the 50-fold increased stability of the TAR-aptamer complex (K(d) = 2.0 nm) over the TAR-TAR* one (K(d) = 92. 5 nm) at physiological concentration of magnesium. Replacement of the wild-type GC pair next to the loop of RNA I' by a GA pair stabilizes the RNA I'-RNA II' loop-loop complex derived from the one involved in the control of the ColE1 plasmid replication. Thus, the GA pair might be the preferred one for stable loop-loop interactions. PMID- 10801858 TI - The active form of the R2F protein of class Ib ribonucleotide reductase from Corynebacterium ammoniagenes is a diferric protein. AB - Corynebacterium ammoniagenes contains a ribonucleotide reductase (RNR) of the class Ib type. The small subunit (R2F) of the enzyme has been proposed to contain a manganese center instead of the dinuclear iron center, which in other class I RNRs is adjacent to the essential tyrosyl radical. The nrdF gene of C. ammoniagenes, coding for the R2F component, was cloned in an inducible Escherichia coli expression vector and overproduced under three different conditions: in manganese-supplemented medium, in iron-supplemented medium, and in medium without addition of metal ions. A prominent typical tyrosyl radical EPR signal was observed in cells grown in rich medium. Iron-supplemented medium enhanced the amount of tyrosyl radical, whereas cells grown in manganese supplemented medium had no such radical. In highly purified R2F protein, enzyme activity was found to correlate with tyrosyl radical content, which in turn correlated with iron content. Similar results were obtained for the R2F protein of Salmonella typhimurium class Ib RNR. The UV-visible spectrum of the C. ammoniagenes R2F radical has a sharp 408-nm band. Its EPR signal at g = 2.005 is identical to the signal of S. typhimurium R2F and has a doublet with a splitting of 0.9 millitesla (mT), with additional hyperfine splittings of 0.7 mT. According to X-band EPR at 77-95 K, the inactive manganese form of the C. ammoniagenes R2F has a coupled dinuclear Mn(II) center. Different attempts to chemically oxidize Mn-R2F showed no relation between oxidized manganese and tyrosyl radical formation. Collectively, these results demonstrate that enzymatically active C. ammoniagenes RNR is a generic class Ib enzyme, with a tyrosyl radical and a diferric metal cofactor. PMID- 10801859 TI - Structural basis for the insensitivity of a serine enzyme (palmitoyl-protein thioesterase) to phenylmethylsulfonyl fluoride. AB - Palmitoyl-protein thioesterase-1 (PPT1) is a newly described lysosomal enzyme that hydrolyzes long chain fatty acids from lipid-modified cysteine residues in proteins. Deficiency in this enzyme results in a severe neurodegenerative storage disorder, infantile neuronal ceroid lipofuscinosis. Although the primary structure of PPT1 contains a serine lipase consensus sequence, the enzyme is insensitive to commonly used serine-modifying reagents phenylmethylsulfonyl fluoride (PMSF) and diisopropylfluorophosphate. In the current paper, we show that the active site serine in PPT1 is modified by a substrate analog of PMSF, hexadecylsulfonylfluoride (HDSF) in a specific and site-directed manner. The apparent K(i) of the inhibition was 125 micrometer (in the presence of 1.5 mm Triton X-100), and the catalytic rate constant for sulfonylation (k(2)) was 3.3/min, a value similar to previously described sulfonylation reactions. PPT1 was crystallized after inactivation with HDSF, and the structure of the inactive form was determined to 2.4 A resolution. The hexadecylsulfonyl was found to modify serine 115 and to snake through a narrow hydrophobic channel that would not accommodate an aromatic sulfonyl fluoride. Therefore, the geometry of the active site accounts for the reactivity of PPT1 with HDSF but not PMSF. These observations suggest a structural explanation as to why certain serine lipases are resistant to modification by commonly used serine-modifying reagents. PMID- 10801860 TI - Cloning, characterization, and phylogenetic analysis of siglec-9, a new member of the CD33-related group of siglecs. Evidence for co-evolution with sialic acid synthesis pathways. AB - The Siglecs are a subfamily of I-type lectins (immunoglobulin superfamily proteins that bind sugars) that specifically recognize sialic acids. We report the cloning and characterization of human Siglec-9. The cDNA encodes a type 1 transmembrane protein with three extracellular immunoglobulin-like domains and a cytosolic tail containing two tyrosines, one within a typical immunoreceptor tyrosine-based inhibitory motif (ITIM). The N-terminal V-set Ig domain has most amino acid residues typical of Siglecs. Siglec-9 is expressed on granulocytes and monocytes. Expression of the full-length cDNA in COS cells induces sialic-acid dependent erythrocyte binding. A recombinant soluble form of the extracellular domain binds to alpha2-3 and alpha2-6-linked sialic acids. Typical of Siglecs, the carboxyl group and side chain of sialic acid are essential for recognition, and mutation of a critical arginine residue in domain 1 abrogates binding. The underlying glycan structure also affects binding, with Galbeta1-4Glc[NAc] being preferred. Siglec-9 shows closest homology to Siglec-7 and both belong to a Siglec-3/CD33-related subset of Siglecs (with Siglecs-5, -6, and -8). The Siglec 9 gene is on chromosome 19q13.3-13.4, in a cluster with all Siglec-3/CD33-related Siglec genes, suggesting their origin by gene duplications. A homology search of the Drosophila melanogaster and Caenorhabditis elegans genomes suggests that Siglec expression may be limited to animals of deuterostome lineage, coincident with the appearance of the genes of the sialic acid biosynthetic pathway. PMID- 10801861 TI - Purification and characterization of rat des-Gln14-Ghrelin, a second endogenous ligand for the growth hormone secretagogue receptor. AB - Ghrelin, a peptide purified from the stomach, is an endogenous ligand for the growth hormone secretagogue receptor (GHS-R) and potently stimulates growth hormone release from the pituitary. Ghrelin is modified with an n-octanoyl group at Ser(3). This modification is essential for the activity of ghrelin. Previously, it was not known whether other ligands for GHS-R existed. Here, we report the purification of the second endogenous ligand for GHS-R from rat stomach. This ligand, named des-Gln(14)-ghrelin, is a 27-amino acid peptide, whose sequence is identical to ghrelin except for one glutamine. Southern blotting analysis under low hybridization conditions indicates that no homologue for ghrelin exists in rat genomic DNA. Furthermore, genomic sequencing and cDNA analysis indicate that des-Gln(14)-ghrelin is not encoded by a gene distinct from ghrelin but is encoded by an mRNA created by alternative splicing of the ghrelin gene. This is the first example of a novel mechanism that produces peptide multiplicity. Des-Gln(14)-ghrelin has an n-octanoyl modification at Ser(3) like ghrelin, which is also essential for its activity. Des-Gln(14)-ghrelin-stimulated growth hormone releases when injected into rats. Thus, growth hormone release is regulated by two gastric peptides, ghrelin and des-Gln(14)-ghrelin. PMID- 10801862 TI - Siglec-9, a novel sialic acid binding member of the immunoglobulin superfamily expressed broadly on human blood leukocytes. AB - Here we characterize the properties and expression pattern of Siglec-9 (sialic acid-binding Ig-like lectin-9), a new member of the Siglec subgroup of the immunoglobulin superfamily. A full-length cDNA encoding Siglec-9 was isolated from a dibutyryl cAMP-treated HL-60 cell cDNA library. Siglec-9 is predicted to contain three extracellular immunoglobulin-like domains that comprise an N terminal V-set domain and two C2-set domains, a transmembrane region and a cytoplasmic tail containing two putative tyrosine-based signaling motifs. Overall, Siglec-9 is approximately 80% identical in amino acid sequence to Siglec 7, suggesting that the genes encoding these two proteins arose relatively recently by gene duplication. Binding assays showed that, similar to Siglec-7, Siglec-9 recognized sialic acid in either the alpha2,3- or alpha2, 6-glycosidic linkage to galactose. Using a specific mAb, Siglec-9 was found to be expressed at high or intermediate levels by monocytes, neutrophils, and a minor population of CD16(+), CD56(-) cells. Weaker expression was observed on approximately 50% of B cells and NK cells and minor subsets of CD8(+) T cells and CD4(+) T cells. These results show that despite their high degree of sequence similarity, Siglec-7 and Siglec-9 have distinct expression profiles. PMID- 10801863 TI - Expression and functional characterization of a Drosophila neuropeptide precursor with homology to mammalian preprotachykinin A. AB - Peptides structurally related to mammalian tachykinins have recently been isolated from the brain and intestine of several insect species, where they are believed to function as both neuromodulators and hormones. Further evidence for the signaling role of insect tachykinin-related peptides was provided by the cloning and characterization of cDNAs for two tachykinin receptors from Drosophila melanogaster. However, no endogenous ligand has been isolated for the Drosophila tachykinin receptors to date. Analysis of the Drosophila genome allowed us to identify a putative tachykinin-related peptide prohormone (prepro DTK) gene. A 1.5-kilobase pair cDNA amplified from a Drosophila head cDNA library contained an 870-base pair open reading frame, which encodes five novel Drosophila tachykinin-related peptides (called DTK peptides) with conserved C terminal FXGXR-amide motifs common to other insect tachykinin-related peptides. The tachykinin-related peptide prohormone gene (Dtk) is both expressed and post translationally processed in larval and adult midgut endocrine cells and in the central nervous system, with midgut expression starting at stage 17 of embryogenesis. The predicted Drosophila tachykinin peptides have potent stimulatory effects on the contractions of insect gut. These data provide additional evidence for the conservation of both the structure and function of the tachykinin peptides in the brain and gut during the course of evolution. PMID- 10801864 TI - An actin subdomain 2 mutation that impairs thin filament regulation by troponin and tropomyosin. AB - Striated muscle thin filaments adopt different quaternary structures, depending upon calcium binding to troponin and myosin binding to actin. Modification of actin subdomain 2 alters troponin-tropomyosin-mediated regulation, suggesting that this region of actin may contain important protein-protein interaction sites. We used yeast actin mutant D56A/E57A to examine this issue. The mutation increased the affinity of tropomyosin for actin 3-fold. The addition of Ca(2+) to mutant actin filaments containing troponin-tropomyosin produced little increase in the thin filament-myosin S1 MgATPase rate. Despite this, three-dimensional reconstruction of electron microscope images of filaments in the presence of troponin and Ca(2+) showed tropomyosin to be in a position similar to that found for muscle actin filaments, where most of the myosin binding site is exposed. Troponin-tropomyosin bound with comparable affinity to mutant and wild type actin in the absence and presence of calcium, and in the presence of myosin S1, tropomyosin bound very tightly to both types of actin. The mutation decreased actin-myosin S1 affinity 13-fold in the presence of troponin-tropomyosin and 2.6 fold in the absence of the regulatory proteins. The results suggest the importance of negatively charged actin subdomain 2 residues 56 and 57 for myosin binding to actin, for tropomyosin-actin interactions, and for regulatory conformational changes in the actin-troponin-tropomyosin complex. PMID- 10801865 TI - Human cationic trypsinogen. Role of Asn-21 in zymogen activation and implications in hereditary pancreatitis. AB - Mutation Asn-21 --> Ile in human cationic trypsinogen (Tg-1) has been associated with hereditary pancreatitis. Recent studies with rat anionic Tg (Tg-2) indicated that the analogous Thr-21 --> Ile mutation stabilizes the zymogen against autoactivation, whereas it has no effect on catalytic properties or autolytic stability of trypsin (Sahin-Toth, M. (1999) J. Biol. Chem. 274, 29699-29704). In the present paper, human cationic Tg (Asn-21-Tg) and mutants Asn-21 --> Ile (Ile 21-Tg) and Asn-21 --> Thr (Thr-21-Tg) were expressed in Escherichia coli, and zymogen activation, zymogen degradation, and trypsin autolysis were studied. Enterokinase activated Asn-21-Tg approximately 2-fold better than Ile-21-Tg or Thr-21-Tg, and catalytic parameters of trypsins were comparable. At 37 degrees C, in 5 mm Ca(2+), all three trypsins were highly stable. In the absence of Ca(2+), Asn-21- and Ile-21-trypsins suffered autolysis in an indistinguishable manner, whereas Thr-21-trypsin exhibited significantly increased stability. In sharp contrast to observations with the rat proenzyme, at pH 8.0, 37 degrees C, autoactivation kinetics of Asn-21-Tg and Ile-21-Tg were identical; however, at pH 5. 0, Ile-21-Tg autoactivated at an enhanced rate relative to Asn-21-Tg. Remarkably, at both pH values, Thr-21-Tg showed markedly higher autoactivation rates than the two other zymogens. Finally, autocatalytic proteolysis of human zymogens was limited to cleavage at Arg-117, and no digestion at Lys-188 was detected. The observations indicate that zymogen stabilization by Ile-21 as observed in rat Tg-2 is not characteristic of human Tg-1. Instead, an increased propensity to autoactivation under acidic conditions might be relevant to the pathomechanism of the Asn-21 --> Ile mutation in hereditary pancreatitis. In the same context, faster autoactivation and increased trypsin stability caused by the Asn-21 --> Thr mutation in human Tg-1 might provide a rationale for the evolutionary divergence from Thr-21 found in other mammalian trypsinogens. PMID- 10801866 TI - Subunit D (Vma8p) of the yeast vacuolar H+-ATPase plays a role in coupling of proton transport and ATP hydrolysis. AB - To investigate the function of subunit D in the vacuolar H(+)-ATPase (V-ATPase) complex, random and site-directed mutagenesis was performed on the VMA8 gene encoding subunit D in yeast. Mutants were selected for the inability to grow at pH 7.5 but the ability to grow at pH 5.5. Mutations leading to reduced levels of subunit D in whole cell lysates were excluded from the analysis. Seven mutants were isolated that resulted in pH-dependent growth but that contained nearly wild type levels of subunit D and nearly normal assembly of the V-ATPase as assayed by subunit A levels associated with isolated vacuoles. Each of these mutants contained 2-3 amino acid substitutions and resulted in loss of 60-100% of proton transport and 58-93% of concanamycin-sensitive ATPase activity. To identify the mutations responsible for the observed effects on activity, 14 single amino acid substitutions and 3 double amino acid substitutions were constructed by site directed mutagenesis and analyzed as described above. Six of the single mutations and all three of the double mutations led to significant (>30%) loss of activity, with the mutations having the greatest effects on activity clustering in the regions Val(71)-Gly(80) and Lys(209)-Met(221). In addition, both M221V and the double mutant V71D/E220V led to significant uncoupling of proton transport and ATPase activity, whereas the double mutant G80D/K209E actually showed increased coupling efficiency. Both a mutant showing reduced coupling and a mutant with only 6% of wild-type proton transport activity showed normal dissociation of the V-ATPase complex in vivo in response to glucose deprivation. These results suggest that subunit D plays an important role in coupling of proton transport and ATP hydrolysis and that only low rates of turnover of the enzyme are required to support in vivo dissociation. PMID- 10801867 TI - Chemically regulated transcription factors reveal the persistence of repressor resistant transcription after disrupting activator function. AB - Control of gene expression often requires that transcription terminates rapidly after destruction, inactivation, or nuclear export of transcription factors. However, the role of transcription factor inactivation in terminating transcription is unclear. We have developed a means of conducting order of addition and co-occupancy experiments in living cells by rapidly exchanging proteins bound to promoters. Using this approach, we found that, following specific disruption of activator function, transcription from active promoters decayed slowly, persisting through multiple cell divisions. This persistent transcriptional activity raised the question of what mechanisms return promoters to inactive states. By exchanging or directing co-occupancy of protein complexes bound to a promoter, we found that the transcriptional inhibitor, Ssn6-Tup1, lost its effectiveness as a repressor following activator dissociation. Similar experiments with another repressor, the histone deacetylase Sin3-Rpd3, reinforced this distinction between repression in the presence and absence of an activator. These results suggest that although repressors such as Ssn6-Tup1 and Sin3-Rpd3 prevent activation of gene expression, other mechanisms of repression return promoters to inactive states following the dissociation or inactivation of a transcriptional activator. PMID- 10801868 TI - Comparing and contrasting Escherichia coli and Mycobacterium tuberculosis mechanosensitive channels (MscL). New gain of function mutations in the loop region. AB - Sequence analysis of 35 putative MscL homologues was used to develop an optimal alignment for Escherichia coli and Mycobacterium tuberculosis MscL and to place these homologues into sequence subfamilies. By using this alignment, previously identified E. coli MscL mutants that displayed severe and very severe gain of function phenotypes were mapped onto the M. tuberculosis MscL sequence. Not all of the resulting M. tuberculosis mutants displayed a gain of function phenotype; for instance, normal phenotypes were noted for mutations at Ala(20), the analogue of the highly sensitive Gly(22) site in E. coli. A previously unnoticed intersubunit hydrogen bond in the extracellular loop region of the M. tuberculosis MscL crystal structure has been analyzed. Cross-linkable residues were substituted for the residues involved in the hydrogen bond, and cross linking studies indicated that these sites are spatially close under physiological conditions. In general, mutation at these positions results in a gain of function phenotype, which provides strong evidence for the importance of the loop region in MscL channel function. No analogue to this interesting interaction could be found in E. coli MscL by sequence alignment. Taken together, these results indicate that caution should be exercised in using the M. tuberculosis MscL crystal structure to analyze previous functional studies of E. coli MscL. PMID- 10801869 TI - Cloning, overexpression, and purification of novobiocic acid synthetase from Streptomyces spheroides NCIMB 11891. AB - Novobiocic acid synthetase, a key enzyme in the biosynthesis of the antibiotic novobiocin, was cloned from the novobiocin producer Streptomyces spheroides NCIMB 11891. The enzyme is encoded by the gene novL, which codes for a protein of 527 amino acids with a calculated mass of 56,885 Da. The protein was overexpressed as a His(6) fusion protein in Escherichia coli and purified to apparent homogeneity by affinity chromatography and gel chromatography. The purified enzyme catalyzed the formation of an amide bond between 3-dimethylallyl-4-hydroxybenzoic acid (ring A of novobiocin) and 3-amino-4,7-dihydroxy-8-methyl coumarin (ring B of novobiocin) in an ATP-dependent reaction. NovL shows homology to the superfamily of adenylate-forming enzymes, and indeed the formation of an acyl adenylate from ring A and ATP was demonstrated by an ATP-PP(i) exchange assay. The purified enzyme exhibited both activation and transferase activity, i.e. it catalyzed both the activation of ring A as acyl adenylate and the subsequent transfer of the acyl group to the amino group of ring B. It is active as a monomer as determined by gel filtration chromatography. The reaction was specific for ATP as nucleotide triphosphate and dependent on the presence of Mg(2+) or Mn(2+). Apparent K(m) values for ring A and ring B were determined as 19 and 131 micrometer respectively. Of several analogues of ring A, only 3-geranyl-4-hydroxybenzoate and to a lesser extent 3-methyl-4-aminobenzoate were accepted as substrates. PMID- 10801870 TI - Biosynthetic origin of hydrogen atoms in the lipase inhibitor lipstatin. AB - The lipase inhibitor lipstatin is biosynthesized in Streptomyces toxytricini via condensation of a C(14) precursor and a C(8) precursor, which are both obtained from fatty acid catabolism. To study the mechanism of this reaction in more detail, S. toxytricini was grown in medium containing a mixture of U-(13)C,U-(2)H lipids and unlabeled sunflower oil or in a medium containing 70% D(2)O. Lipstatin was isolated and analyzed by (1)H,(2)H, and (13)C NMR spectroscopy. Hydrogen atoms at C-2, C-3, and C-4 of lipstatin were found to be derived from solvent protons. The formation of the lipstatin precursor 3-hydroxy-Delta(5,8) tetradecadienoyl-CoA by beta oxidation of linoleic acid explains the incorporation of solvent hydrogen into the 4 position of lipstatin. The hydrogen in position 3 of lipstatin is most probably introduced from solvent by proton/deuterium exchange of a redox cofactor involved in the reduction of the keto group in the branched chain beta keto acid arising by a decarboxylative condensation. The incorporation of solvent hydrogen at position 2 can be explained by epimerization of a chiral intermediate at C-2 and C-3. Epimerization may involve a dehydration-rehydration mechanism. PMID- 10801871 TI - Feedback inhibition of sodium/calcium exchange by mitochondrial calcium accumulation. AB - Chinese hamster ovary cells expressing the bovine cardiac Na(+)/Ca(2+) exchanger were subjected to two periods of 5 and 3 min, respectively, during which the extracellular Na(+) concentration ([Na(+)](o)) was reduced to 20 mm; these intervals were separated by a 5-min recovery period at 140 mm Na(+)(o). The cytosolic Ca(2+) concentration ([Ca(2+)](i)) increased during both intervals due to Na(+)-dependent Ca(2+) influx by the exchanger. However, the peak rise in [Ca(2+)](i) during the second interval was only 26% of the first. The reduced rise in [Ca(2+)](i) was due to an inhibition of Na(+)/Ca(2+) exchange activity rather than increased Ca(2+) sequestration since the influx of Ba(2+), which is not sequestered by internal organelles, was also inhibited by a prior interval of Ca(2+) influx. Mitochondria accumulated Ca(2+) during the first interval of reduced [Na(+)](o), as determined by an increase in fluorescence of the Ca(2+) indicating dye rhod-2, which preferentially labels mitochondria. Agents that blocked mitochondrial Ca(2+) accumulation (uncouplers, nocodazole) eliminated the observed inhibition of exchange activity during the second period of low [Na(+)](o). Conversely, diltiazem, an inhibitor of the mitochondrial Na(+)/Ca(2+) exchanger, increased mitochondrial Ca(2+) accumulation and also increased the inhibition of exchange activity. We conclude that Na(+)/Ca(2+) exchange activity is regulated by a feedback inhibition process linked to mitochondrial Ca(2+) accumulation. PMID- 10801872 TI - Maturation and pro-peptide cleavage of beta-secretase. AB - Amyloid beta-peptide is generated by two sequential proteolytic cleavages mediated by beta-secretase (BACE) and gamma-secretase. BACE was recently identified as a membrane-associated aspartyl protease. We have now analyzed the maturation and pro-peptide cleavage of BACE. Pulse-chase experiments revealed that BACE is post-translationally modified during transport to the cell surface, which can be monitored by a significant increase in the molecular mass. The increase in molecular mass is caused by complex N-glycosylation. Treatment with tunicamycin and N-glycosidase F led to a BACE derivative with a molecular weight corresponding to an unmodified version. In contrast, the mature form of BACE was resistant to endoglycosidase H treatment. The cytoplasmic tail of BACE was required for efficient maturation and trafficking through the Golgi; a BACE variant lacking the cytoplasmic tail undergoes inefficient maturation. In contrast a soluble BACE variant that does not contain a membrane anchor matured more rapidly than full-length BACE. Pro-BACE was predominantly located within the endoplasmic reticulum. Pro-peptide cleavage occurred immediately before full maturation and trafficking through the Golgi. PMID- 10801873 TI - Raf-1-associated protein phosphatase 2A as a positive regulator of kinase activation. AB - The Raf-1 kinase plays a key role in relaying proliferation signals elicited by mitogens or oncogenes. Raf-1 is regulated by complex and incompletely understood mechanisms including phosphorylation. A number of studies have indicated that phosphorylation of serines 259 and 621 can inhibit the Raf-1 kinase. We show that both serines are hypophosphorylated during early mitogenic stimulation and that hypophosphorylation correlates with peak Raf-1 activation. Concentrations of okadaic acid that selectively inhibit protein phosphatase 2A (PP2A) induce phosphorylation of these residues and prevent maximal activation of the Raf-1 kinase. This effect is mediated via phosphorylation of serine 259. The PP2A core heterodimer forms complexes with Raf-1 in vivo and in vitro. These data identify PP2A as a positive regulator of Raf-1 activation and are the first indication that PP2A may support the activation of an associated kinase. PMID- 10801874 TI - Pyridoxamine, an inhibitor of advanced glycation reactions, also inhibits advanced lipoxidation reactions. Mechanism of action of pyridoxamine. AB - Maillard or browning reactions lead to formation of advanced glycation end products (AGEs) on protein and contribute to the increase in chemical modification of proteins during aging and in diabetes. AGE inhibitors such as aminoguanidine and pyridoxamine (PM) have proven effective in animal model and clinical studies as inhibitors of AGE formation and development of diabetic complications. We report here that PM also inhibits the chemical modification of proteins during lipid peroxidation (lipoxidation) reactions in vitro, and we show that it traps reactive intermediates formed during lipid peroxidation. In reactions of arachidonate with the model protein RNase, PM prevented modification of lysine residues and formation of the advanced lipoxidation end products (ALEs) N(epsilon)-(carboxymethyl)lysine, N(epsilon)-(carboxyethyl)lysine, malondialdehyde-lysine, and 4-hydroxynonenal-lysine. PM also inhibited lysine modification and formation of ALEs during copper-catalyzed oxidation of low density lipoprotein. Hexanoic acid amide and nonanedioic acid monoamide derivatives of PM were identified as major products formed during oxidation of linoleic acid in the presence of PM. We propose a mechanism for formation of these products from the 9- and 13-oxo-decadienoic acid intermediates formed during peroxidation of linoleic acid. PM, as a potent inhibitor of both AGE and ALE formation, may prove useful for limiting the increased chemical modification of tissue proteins and associated pathology in aging and chronic diseases, including both diabetes and atherosclerosis. PMID- 10801875 TI - A carboxyl-terminal region important for the expression and targeting of the skeletal muscle dihydropyridine receptor. AB - We have used the yeast two-hybrid technique and expression of truncated/mutated dihydropyridine receptors (DHPRs) to investigate whether the carboxyl tail of the DHPR is involved in targeting to junctions between the sarcolemma and sarcoplasmic reticulum in skeletal muscle. The carboxyl tail was extremely reactive in yeast two-hybrid library screens, with the reactivity residing in amino acids 1621-1647 and abolished by a point mutation (V1642D). Dysgenic myotubes were injected with cDNA encoding green fluorescent protein fused to the amino terminus of DHPRs truncated after either residue 1620 (Delta1621-1873) or residue 1542 (Delta1543-1873) or of full-length DHPRs with the V1642D mutation (V1642D). For either Delta1621-1873 or V1642D, the restoration of excitation contraction coupling was reduced approximately 40%, and the number of functional DHPRs in the sarcolemma was reduced approximately 30%, compared with the wild type DHPR. The restoration of excitation-contraction coupling and surface expression was more drastically reduced (by approximately 90 and approximately 55%, respectively) for Delta1543-1873. Fluorescence microscopy revealed that Delta1621-1873 and V1642D were concentrated in a longitudinally restricted region near the injected nucleus, whereas wild-type DHPRs were present relatively uniformly along the length of a myotube. The intensity of fluorescence was greatly reduced for Delta1543-1873, indicating a low level of protein expression. Thus, residues 1543-1647 appear to play a role in the biosynthetic processing, transport, and/or anchoring of DHPRs, with residues 1543-1620 being particularly important for expression. PMID- 10801876 TI - The Asn-420-linked sugar chain in human epidermal growth factor receptor suppresses ligand-independent spontaneous oligomerization. Possible role of a specific sugar chain in controllable receptor activation. AB - To elucidate a role(s) of Asn-linked sugar chain(s) in the function of epidermal growth factor receptor (EGFR), a series of the EGFR mutants were prepared in which potential glycosylation sites in the domain III were eliminated by site directed mutagenesis. Although the wild-type and mutants of Asn-328, Asn-337, and Asn-389 underwent autophosphorylation in response to epidermal growth factor (EGF), the Asn-420 --> Gln mutant was found to be constitutively tyrosine phosphorylated. This abnormal ligand-independent phosphorylation of the mutant appears to be due to a ligand-independent spontaneous oligomer formation, as shown by a cross-linking experiment using the purified soluble extracellular domain (sEGFR). As revealed by the dissociation of the Asn-420 --> Gln sEGFR oligomer by simple dilution, it seems likely that the equilibrium is shifted toward oligomer formation to an unusual degree. Furthermore, it was also found that the mutation caused a loss of the ability to bind EGF. These findings suggest that the sugar chain linked to Asn-420 plays a crucial role in EGF binding and prevents spontaneous oligomerization of the EGFR, which may otherwise lead to uncontrollable receptor activation, and support the view of a specific role of an Asn-linked sugar chain in the function of a glycoprotein. PMID- 10801877 TI - Conformational reorganization of the four-helix bundle of human apolipoprotein E in binding to phospholipid. AB - Conformational reorganization of the amino-terminal four-helix bundle (22-kDa fragment) of apolipoprotein E (apoE) in binding to the phospholipid dimyristoylphosphatidylcholine (DMPC) to form discoidal particles was investigated by introducing single, double, and triple interhelical disulfide bonds to restrict the opening of the bundle. Interaction of apoE with DMPC was assessed by vesicle disruption, turbidimetric clearing, and gel filtration assays. The results indicate that the formation of apoE.DMPC discoidal particles occurs in a series of steps. A triple disulfide mutant, in which all four helices were tethered, did not form complexes but could release encapsulated 5-(6) carboxylfluorescein from DMPC vesicles, indicating that the initial interaction does not involve major reorganization of the helical bundle. Initial interaction is followed by the opening of the four-helix bundle to expose the hydrophobic faces of the amphipathic helices. In this step, helices 1 and 2 and helices 3 and 4 preferentially remain paired, since these disulfide-linked mutants bound to DMPC in a manner similar to that of the 22-kDa fragment of apoE4. In contrast, mutants in which helices 2 and 3 and/or helices 1 and 4 paired bound poorly to DMPC. However, all single and double helical pairings resulted in the formation of larger discs than were formed by the 22-kDa fragment, indicating that further reorganization of the helices occurs following the initial opening of the four helix bundle in which the protein assumes its final lipid-bound conformation. In support of this rearrangement, reducing the disulfide bonds converted the large disulfide mutant discs to normal size. PMID- 10801878 TI - Why expression of phosphatidylethanolamine N-methyltransferase does not rescue Chinese hamster ovary cells that have an impaired CDP-choline pathway. AB - The mutant Chinese hamster ovary cell line (CHO), MT58, has a temperature sensitive mutation in CTP:phosphocholine cytidylyltransferase (CT), preventing phosphatidylcholine (PC) synthesis at 40 degrees C which results in apoptosis. Previous studies (Houweling, M., Cui, Z., and Vance, D. E. (1995) J. Biol. Chem. 270, 16277-16282) showed that expression of wild-type CT-alpha rescued the cells at 40 degrees C, whereas expression of phosphatidylethanolamine N methyltransferase-2 (PEMT2) did not, even though PC levels appeared to be maintained at wild-type levels after 24 h at the restrictive temperature. We report that the failure of PEMT2 to rescue the MT58 cell line is due to inadequate long term PC synthesis. We found that changing the medium every 24 h rescued the PEMT2-expressing MT58 cells grown at 40 degrees C. This was due to the uptake and utilization of lipids in the serum. At 40 degrees C, PC levels in the wild-type CHO cells and CT-expressing MT58 cells increased over time whereas PC levels did not change in both the MT58 and PEMT2-expressing MT58 cell lines. Further investigation found that both the PEMT2-expressing MT58 and MT58 cell lines accumulated triacylglycerol at 40 degrees C. Pulse-chase experiments indicated that lyso-PC accumulated to a higher degree at 40 degrees C in the PEMT2-expressing MT58 cells compared with CT-expressing MT58 cells. Transfection of the PEMT-expressing MT58 cells with additional PEMT2 cDNA partially rescued the growth of these cells at 40 degrees C. Inhibition of PC degradation, by inhibitors of phospholipases, also stimulated PEMT-expressing MT58 cell growth at 40 degrees C. Best results were observed using a calcium-independent phospholipase A(2) inhibitor, methyl arachidonyl fluorophosphonate. This inhibitor also increased PC mass in the PEMT2-expressing MT58 cells. When the cells are shifted to 40 degrees C, PC degradation by enzymes such as phospholipases is greater than PC synthesis in the mutant PEMT2-expressing MT58 cells. Taken together, these results indicate that PEMT2 expression fails to rescue the mutant cell line at 40 degrees C because it does not maintain PC levels required for cellular replication. PMID- 10801879 TI - Characterization of Drosophila insulin receptor substrate. AB - Insulin receptor substrate (IRS) proteins are phosphorylated by multiple tyrosine kinases, including the insulin receptor. Phosphorylated IRS proteins bind to SH2 domain-containing proteins, thereby triggering downstream signaling pathways. The Drosophila insulin receptor (dIR) C-terminal extension contains potential binding sites for signaling molecules, suggesting that dIR might not require an IRS protein to accomplish its signaling functions. However, we obtained a cDNA encoding Drosophila IRS (dIRS), and we demonstrated expression of dIRS in a Drosophila cell line. Like mammalian IRS proteins, the N-terminal portion of dIRS contains a pleckstrin homology domain and a phosphotyrosine binding domain that binds to phosphotyrosine residues in both human and Drosophila insulin receptors. When coexpressed with dIRS in COS-7 cells, a chimeric receptor (the extracellular domain of human IR fused to the cytoplasmic domain of dIR) mediated insulin stimulated tyrosine phosphorylation of dIRS. Mutating the juxtamembrane NPXY motif markedly reduced the ability of the receptor to phosphorylate dIRS. In contrast, the NPXY motifs in the C-terminal extension of dIR were required for stable association with dIRS. Coimmunoprecipitation experiments demonstrated insulin-dependent binding of dIRS to phosphatidylinositol 3-kinase and SHP2. However, we did not detect interactions with Grb2, SHC, or phospholipase C-gamma. Taken together with published genetic studies, these biochemical data support the hypothesis that dIRS functions directly downstream from the insulin receptor in Drosophila. PMID- 10801880 TI - The destabilization of lipid membranes induced by the C-terminal fragment of caspase 8-cleaved bid is inhibited by the N-terminal fragment. AB - Bid is a proapoptotic, BH3-domain-only member of the Bcl-2 family. In Fas-induced apoptosis, Bid is activated through cleavage by caspase 8 into a 15.5-kDa C terminal fragment (t(c)Bid) and a 6.5 kDa N-terminal fragment (t(n)Bid). Following the cleavage, t(c)Bid translocates to the mitochondria and promotes the release of cytochrome c into the cytosol by a mechanism that is not understood. Here we report that recombinant t(c)Bid can act as a membrane destabilizing agent. t(c)Bid induces destabilization and breaking of planar lipid bilayers without appearance of ionic channels; its destabilizing activity is comparable with that of Bax and at least 30-fold higher than that of full-length Bid. Consistently, t(c)Bid, but not full-length Bid, permeabilizes liposomes at physiological pH. The destabilizing effect of t(c)Bid on liposomes and planar bilayers is independent of the BH3 domain. In contrast, mutations in the BH3 domain impair t(c)Bid ability to induce cytochrome c release from mitochondria. The permeabilizing effect of t(c)Bid on planar bilayers, liposomes, and mitochondria can be inhibited by t(n)Bid. In conclusion, our results suggest a dual role for Bid: BH3-independent membrane destabilization and BH3-dependent interaction with other proteins. Moreover, the dissociation of Bid after cleavage by caspase 8 represents an additional step at which apoptosis may be regulated. PMID- 10801881 TI - Identity of urinary trypsin inhibitor-binding protein to link protein. AB - Urinary trypsin inhibitor (UTI), a Kunitz-type protease inhibitor, directly binds to some types of cells via cell-associated UTI-binding proteins (UTI-BPs). Here we report that the 40-kDa protein (UTI-BP(40)) was purified from the cultured human chondrosarcoma cell line HCS-2/8 by UTI affinity chromatography. Purified UTI-BP(40) was digested with trypsin, and the amino acid sequences of the peptide fragments were determined. The sequences of six tryptic fragments of UTI-BP(40) were identical to subsequences present in human link protein (LP). Authentic bovine LP and UTI-BP(40) displayed identical electrophoretic and chromatographic behavior. The UTI-binding properties of UTI-BP(40) and LP were indistinguishable. Direct binding and competition studies strongly demonstrated that the NH(2) terminal fragment is the UTI-binding part of the LP molecule, that the COOH terminal UTI fragment (HI-8) failed to bind the NH(2)-terminal subdomain of the LP molecule, and that LP and UTI-BP(40) exhibited significant hyaluronic acid binding. These results demonstrate that UTI-BP(40) is identical to LP and that the NH(2)-terminal domain of UTI is involved in the interaction with the NH(2) terminal fragment of LP, which is bound to hyaluronic acid in the extracellular matrix. PMID- 10801882 TI - Mapping and regulation of the tumor-associated epitope recognized by monoclonal antibody RS-11. AB - We have previously described a rat monoclonal antibody, RS-11, which recognizes a tumor-associated antigen common to several species. In the present study, we have cloned and characterized the antigen recognized by RS-11. We screened a phage expression library prepared from HeLa cDNA and identified a clone that reacts with RS-11. DNA sequence analysis revealed that this clone contains sequences of keratin 18 (nucleotides 568-1196). We constructed several glutathione S transferase fusion proteins and synthetic peptides based on this DNA sequence analysis and examined their reactivity with RS-11 to accurately map the RS-11 epitope. We determined that the epitope resides within a region of seven amino acids on the alpha-helix 2B domain of keratin 18 in which two amino acids (Leu(366) and Lys(370)) are completely conserved among intermediate filaments as well as other keratin members that are immunoreactive with RS-11. These two residues are sequentially discontinuous but spatially adjacent. The RS-11 epitope is constitutively present in human primary cultured hepatocytes; however, its immunoreactivity with RS-11 is up-regulated by malignant transformation or stimulation with either epidermal growth factor or transforming growth factor alpha. PMID- 10801883 TI - Regulatory domain conformational exchange and linker region flexibility in cardiac troponin C bound to cardiac troponin I. AB - Previously, we utilized (15)N transverse relaxation rates to demonstrate significant mobility in the linker region and conformational exchange in the regulatory domain of Ca(2+)-saturated cardiac troponin C bound to the isolated N domain of cardiac troponin I (Gaponenko, V., Abusamhadneh, E., Abbott, M. B., Finley, N., Gasmi-Seabrook, G., Solaro, R.J., Rance, M., and Rosevear, P.R. (1999) J. Biol. Chem. 274, 16681-16684). Here we show a large decrease in cardiac troponin C linker flexibility, corresponding to residues 85-93, when bound to intact cardiac troponin I. The addition of 2 m urea to the intact cardiac troponin I-troponin C complex significantly increased linker flexibility. Conformational changes in the regulatory domain of cardiac troponin C were monitored in complexes with troponin I-(1-211), troponin I-(33-211), troponin I (1-80) and bisphosphorylated troponin I-(1-80). The cardiac specific N terminus, residues 1-32, and the C-domain, residues 81-211, of troponin I are both capable of inducing conformational changes in the troponin C regulatory domain. Phosphorylation of the cardiac specific N terminus reversed its effects on the regulatory domain. These studies provide the first evidence that the cardiac specific N terminus can modulate the function of troponin C by altering the conformational equilibrium of the regulatory domain. PMID- 10801884 TI - Multiple roles for the twin arginine leader sequence of dimethyl sulfoxide reductase of Escherichia coli. AB - Dimethyl sulfoxide (Me(2)SO) reductase of Escherichia coli is a terminal electron transport chain enzyme that is expressed under anaerobic growth conditions and is required for anaerobic growth with Me(2)SO as the terminal electron acceptor. The trimeric enzyme is composed of a membrane extrinsic catalytic dimer (DmsAB) and a membrane intrinsic anchor (DmsC). The amino terminus of DmsA has a leader sequence with a twin arginine motif that targets DmsAB to the membrane via a novel Sec-independent mechanism termed MTT for membrane targeting and translocation. We demonstrate that the Met-1 present upstream of the twin arginine motif serves as the correct translational start site. The leader is essential for the expression of DmsA, stability of the DmsAB dimer, and membrane targeting of the reductase holoenzyme. Mutation of arginine 17 to aspartate abolished membrane targeting. The reductase was labile in the leader sequence mutants. These mutants failed to support growth on glycerol-Me(2)SO minimal medium. Replacing the DmsA leader with the TorA leader of trimethylamine N-oxide reductase produced a membrane-bound DmsABC with greatly reduced enzyme activity and inefficient anaerobic respiration indicating that the twin arginine leaders may play specific roles in the assembly of redox enzymes. PMID- 10801885 TI - Microtubules regulate local Ca2+ spiking in secretory epithelial cells. AB - The role of the cytoskeleton in regulating Ca(2+) release has been explored in epithelial cells. Trains of local Ca(2+) spikes were elicited in pancreatic acinar cells by infusion of inositol trisphosphate through a whole cell patch pipette, and the Ca(2+)-dependent Cl(-) current spikes were recorded. The spikes were only transiently inhibited by cytochalasin B, an agent that acts on microfilaments. In contrast, nocodazole (5-100 micrometer), an agent that disrupts the microtubular network, dose-dependently reduced spike frequency and decreased spike amplitude leading to total blockade of the response. Consistent with an effect of microtubular disruption, colchicine also inhibited spiking but neither Me(2)SO nor beta-lumicolchicine, an inactive analogue of colchicine, had any effect. The microtubule-stabilizing agent, taxol, also inhibited spiking. The nocodazole effects were not due to complete loss of function of the Ca(2+) signaling apparatus, because supramaximal carbachol concentrations were still able to mobilize a Ca(2+) response. Finally, as visualized by 2-photon excitation microscopy of ER-Tracker, nocodazole promoted a loss of the endoplasmic reticulum in the secretory pole region. We conclude that microtubules specifically maintain localized Ca(2+) spikes at least in part because of the local positioning of the endoplasmic reticulum. PMID- 10801886 TI - Promoter sequences of the putative Anopheles gambiae apyrase confer salivary gland expression in Drosophila melanogaster. AB - The saliva of blood-feeding arthropods contains an apyrase that facilitates hematophagy by inhibiting the ADP-induced aggregation of the host platelets. We report here the isolation of a salivary gland-specific cDNA encoding a secreted protein that likely represents the Anopheles gambiae apyrase. We describe also two additional members of the apyrase/5'-nucleotidase family. The cDNA corresponding to the AgApyL1 gene encodes a secreted protein that is closely related in sequence to the apyrase of the yellow fever mosquito, Aedes aegypti, and whose expression appears enriched in, but not restricted to, female salivary glands. The AgApyL2 gene was found searching an A. gambiae data base, and its expression is restricted to larval stages. We isolated the gene encoding the presumed A. gambiae apyrase (AgApy) and we tested its putative promoter for the tissue-specific expression of the LacZ gene from Escherichia coli in transgenic Drosophila melanogaster. All the transgenic lines analyzed showed a weak but unambiguous staining of the adult glands, indicating that some of the salivary gland-specific transcriptional regulatory elements are conserved between the malaria mosquito and the fruit fly. The availability of salivary gland-specific promoters may be useful both for studies on vector-parasite interactions and, potentially, for the targeted tissue-specific expression of anti-parasite genes in the mosquito. PMID- 10801887 TI - Brain-enriched hyaluronan binding (BEHAB)/brevican cleavage in a glioma cell line is mediated by a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) family member. AB - Brain-enriched hyaluronan binding (BEHAB)/brevican is a brain-specific extracellular matrix protein containing a cleavage site between Glu(395) Ser(396), which bears remarkable homology to the "aggrecanase" site in the cartilage proteoglycan aggrecan. Expression of BEHAB/brevican is dramatically increased in human gliomas, notoriously invasive tumors. Recently, we showed that the rat 9L gliosarcoma cell line, which does not express BEHAB/brevican and forms non-invasive tumors when grown as intracranial grafts, can form invasive tumors when transfected with a 5' cDNA fragment of BEHAB/brevican, but not when transfected with the full-length cDNA. In marked contrast, the highly invasive CNS-1 glioma cell line expresses and cleaves BEHAB/brevican protein when grown as an intracranial graft. These results suggest that both synthesis and cleavage of BEHAB/brevican protein may play a role in the invasiveness of gliomas. We report here, using an antibody developed to the neoepitope created by BEHAB/brevican cleavage at the Glu(395)-Ser(396) site, that the CNS-1 cells are able to cleave the protein in vitro. We characterized the CNS-1-derived cleavage activity by assaying its ability to cleave BEHAB/brevican proteoglycan, and determined that the enzyme is a constitutively expressed, secreted activity. Using a variety of protease inhibitors, reverse transcriptase-polymerase chain reaction, and specific antibodies, we determined that this activity is likely to be a member of the ADAMTS family of metalloproteinases, specifically ADAMTS4. These results suggest a novel function for ADAMTS family members in BEHAB/brevican cleavage and glioma and indicate that inhibition of ADAMTS in glioma may provide a novel therapeutic strategy. PMID- 10801888 TI - Binding of PurH to a muscle-specific splicing enhancer functionally correlates with exon inclusion in vivo. AB - Regulated alternative splicing of avian cardiac troponin T (cTNT) pre-mRNA requires multiple intronic elements called muscle-specific splicing enhancers (MSEs) that flank the alternative exon 5 and promote muscle-specific exon inclusion. To understand the function of the MSEs in muscle-specific splicing, we sought to identify trans-acting factors that bind to these elements. MSE3, which is located 66-81 nucleotides downstream of exon 5, assembles a complex that is both sequence- and muscle-specific. Purification and characterization of the MSE3 complex identified one component as 5-aminoimidazole-4-carboxamide ribonucleotideformyltransferase/IMP cyclohydrolase (PurH), an enzyme involved in de novo purine synthesis. Recombinant human PurH protein directly binds MSE3 RNA and PurH is the primary determinant of sequence-specific binding in the native complex. Furthermore, we show a direct correlation between the in vitro binding affinity of both the MSE3 complex and recombinant PurH with functional activation of exon inclusion in vivo. Together, these results strongly suggest that PurH performs a second function as a component of a complex that regulates MSE3 dependent exon inclusion. PMID- 10801889 TI - Histone H2A.Z is widely but nonrandomly distributed in chromosomes of Drosophila melanogaster. AB - Variant histones that differ in amino acid sequence from S-phase histones are widespread in eukaryotes, yet the structural changes they cause to nucleosomes and how those changes affect relevant cellular processes have not been determined. H2A.F/Z is a highly conserved family of H2A variants. H2Av, the H2A.F/Z variant of Drosophila melanogaster, was localized in polytene chromosomes by indirect immunofluorescence and in diploid chromosomes by chromatin immunoprecipitation. H2Av was widely distributed in the genome and not limited to sites of active transcription. H2Av was present in thousands of euchromatic bands and the heterochromatic chromocenter of polytene chromosomes, and the H2Av antibody precipitated both transcribed and nontranscribed genes as well as noncoding euchromatic and heterochromatic sequences. The distribution of H2Av was not uniform. The complex banding pattern of H2Av in polytene chromosomes did not parallel the concentration of DNA, as did the pattern of immunofluorescence using H2A antibodies, and the density of H2Av measured by immunoprecipitation varied between different sequences. Of the sequences assayed, H2Av was least abundant on 1. 688 satellite sequences and most abundant on the hsp70 genes. Finally, transcription caused, to an equivalent extent, both H2Av and H2A to be less tightly associated with DNA. PMID- 10801890 TI - Identification and functional expression of four isoforms of ATPase II, the putative aminophospholipid translocase. Effect of isoform variation on the ATPase activity and phospholipid specificity. AB - ATPase II, a vanadate-sensitive and phosphatidylserine-dependent Mg(2+)-ATPase, is a member of a subfamily of P-type ATPase and is presumably responsible for aminophospholipid translocation activity in eukaryotic cells. The aminophospholipid translocation activity plays an important physiological role in the maintenance of membrane phospholipid asymmetry that is observed in the plasma membrane as well as the membranes of certain cellular organelles. While the preparations of ATPase II from different sources share common fundamental properties, such as substrate specificity, inhibitor spectrum, and phospholipid dependence, they are divergent in several characteristics. These include specific ATPase activity and phospholipid selectivity. We report here the identification of four isoforms of ATPase II in bovine brain. These isoforms are formed by a combination of two major variations in their primary sequences and show that the structural variation of these isoforms has functional significance in both ATPase activity and phosholipid selectivity. Furthermore, studies with the phosphoenzyme intermediate of ATPase II and its recombinant isoforms revealed that phosphatidylserine is essential for the dephosphorylation of the intermediate. Without phosphatidylserine, ATPase II would be accumulated as phosphoenzyme in the presence of ATP, resulting in the interruption of its catalytic cycle. PMID- 10801892 TI - Identification and molecular characterization of the first alpha -xylosidase from an archaeon. AB - We here report the first molecular characterization of an alpha-xylosidase (XylS) from an Archaeon. Sulfolobus solfataricus is able to grow at temperatures higher than 80 degrees C on several carbohydrates at acidic pH. The isolated xylS gene encodes a monomeric enzyme homologous to alpha-glucosidases, alpha-xylosidases, glucoamylases and sucrase-isomaltases of the glycosyl hydrolase family 31. xylS belongs to a cluster of four genes in the S. solfataricus genome, including a beta-glycosidase, an hypothetical membrane protein homologous to the major facilitator superfamily of transporters, and an open reading frame of unknown function. The alpha-xylosidase was overexpressed in Escherichia coli showing optimal activity at 90 degrees C and a half-life at this temperature of 38 h. The purified enzyme follows a retaining mechanism of substrate hydrolysis, showing high hydrolytic activity on the disaccharide isoprimeverose and catalyzing the release of xylose from xyloglucan oligosaccharides. Synergy is observed in the concerted in vitro hydrolysis of xyloglucan oligosaccharides by the alpha xylosidase and the beta-glycosidase from S. solfataricus. The analysis of the total S. solfataricus RNA revealed that all the genes of the cluster are actively transcribed and that xylS and orf3 genes are cotranscribed. PMID- 10801893 TI - Purification and characterization of ACR2p, the Saccharomyces cerevisiae arsenate reductase. AB - In Saccharomyces cerevisiae, expression of the ACR2 and ACR3 genes confers arsenical resistance. Acr2p is the first identified eukaryotic arsenate reductase. It reduces arsenate to arsenite, which is then extruded from cells by Acr3p. In this study, we demonstrate that ACR2 complemented the arsenate sensitive phenotype of an arsC deletion in Escherichia coli. ACR2 was cloned into a bacterial expression vector and expressed in E. coli as a C-terminally histidine-tagged protein that was purified by sequential metal chelate affinity and gel filtration chromatography. Acr2p purified as a homodimer of 34 kDa. The purified protein was shown to catalyze the reduction of arsenate to arsenite. Enzymatic activity as a function of arsenate concentration exhibited an apparent positive cooperativity with an apparent Hill coefficient of 2.7. Activity required GSH and glutaredoxin as the source of reducing equivalents. Thioredoxin was unable to support arsenate reduction. However, glutaredoxins from both S. cerevisiae and E. coli were able to serve as reductants. Analysis of grx mutants lacking one or both cysteine residues in the Cys-Pro-Tyr-Cys active site demonstrated that only the N-terminal cysteine residue is essential for arsenate reductase activity. This suggests that during the catalytic cycle, Acr2p forms a mixed disulfide with GSH before being reduced by glutaredoxin to regenerate the active Acr2p reductase. PMID- 10801894 TI - Peroxynitrite targets the epidermal growth factor receptor, Raf-1, and MEK independently to activate MAPK. AB - Activation of ERK-1 and -2 by H(2)O(2) in a variety of cell types requires epidermal growth factor receptor (EGFR) phosphorylation. In this study, we investigated the activation of ERK by ONOO(-) in cultured rat lung myofibroblasts. Western blot analysis using anti-phospho-ERK antibodies along with an ERK kinase assay using the phosphorylated heat- and acid-stable protein (PHAS-1) substrate demonstrated that ERK activation peaked within 15 min after ONOO(-) treatment and was maximally activated with 100 micrometer ONOO(-). Activation of ERK by ONOO(-) and H(2)O(2) was blocked by the antioxidant N-acetyl l-cysteine. Catalase blocked ERK activation by H(2)O(2), but not by ONOO(-), demonstrating that the effect of ONOO(-) was not due to the generation of H(2)O(2). Both H(2)O(2) and ONOO(-) induced phosphorylation of EGFR in Western blot experiments using an anti-phospho-EGFR antibody. However, the EGFR tyrosine kinase inhibitor AG1478 abolished ERK activation by H(2)O(2), but not by ONOO(-). Both H(2)O(2) and ONOO(-) activated Raf-1. However, the Raf inhibitor forskolin blocked ERK activation by H(2)O(2), but not by ONOO(-). The MEK inhibitor PD98059 inhibited ERK activation by both H(2)O(2) and ONOO(-). Moreover, ONOO(-) or H(2)O(2) caused a cytotoxic response of myofibroblasts that was prevented by preincubation with PD98059. In a cell-free kinase assay, ONOO(-) (but not H(2)O(2)) induced autophosphorylation and nitration of a glutathione S transferase-MEK-1 fusion protein. Collectively, these data indicate that ONOO(-) activates EGFR and Raf-1, but these signaling intermediates are not required for ONOO(-)-induced ERK activation. However, MEK-1 activation is required for ONOO(-) induced ERK activation in myofibroblasts. In contrast, H(2)O(2)-induced ERK activation is dependent on EGFR activation, which then leads to downstream Raf-1 and MEK-1 activation. PMID- 10801895 TI - Peroxisome proliferator-activated receptor gamma ligands inhibit retinoblastoma phosphorylation and G1--> S transition in vascular smooth muscle cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily that is activated by binding certain fatty acids, eicosanoids, and insulin-sensitizing thiazolidinediones (TZD). The TZD troglitazone (TRO) inhibits vascular smooth muscle cell proliferation and migration both in vitro and in vivo. The precise mechanism of its antiproliferative activity, however, has not been elucidated. We report here that PPARgamma ligands inhibit rat aortic vascular smooth muscle cell proliferation by blocking the events critical for G(1) --> S progression. Flow cytometry demonstrated that both TRO and another TZD, rosiglitazone, prevented G(1) --> S progression induced by platelet-derived growth factor and insulin. Movement of cells from G(1) --> S was also inhibited by the non-TZD, natural PPARgamma ligand 15-deoxy-(12,14)Delta prostaglandin J(2) (15d-PGJ(2)), and the mitogen-activated protein kinase pathway inhibitor PD98059. Inhibition of G(1) --> S exit by these compounds was accompanied by a substantial blockade of retinoblastoma protein phosphorylation. TRO and rosiglitazone attenuated both the mitogen-induced degradation of p27(kip1) and the mitogenic induction of p21(cip1). 15d-PGJ(2) and PD98059 inhibited both the degradation of p27(kip1) and the induction of cyclin D1 in response to mitogens. These effects resulted in the inhibition of mitogenic stimulation of cyclin-dependent kinases activated by cyclins D1 and E. These data demonstrate that PPARgamma ligands are antiproliferative drugs that act by modulating cyclin-dependent kinase inhibitors; they may provide a new therapeutic approach for proliferative vascular diseases. PMID- 10801896 TI - Integrin modulation by lateral association. PMID- 10801897 TI - Ligand binding to integrins. PMID- 10801898 TI - The leukocyte integrins. PMID- 10801899 TI - Integrins and actin filaments: reciprocal regulation of cell adhesion and signaling. PMID- 10801900 TI - Regulation of biological nitrogen fixation. AB - Biological nitrogen fixation, a process found only in some prokaryotes, is catalyzed by the nitrogenase enzyme complex. Bacteria containing nitrogenase occupy an indispensable ecological niche, supplying fixed nitrogen to the global nitrogen cycle. Due to this inceptive role in the nitrogen cycle, diazotrophs are present in virtually all ecosystems, with representatives in environments as varied as aerobic soils (e.g., Azotobacter species), the ocean surface layer (Trichodesmium) and specialized nodules in legume roots (Rhizobium). In any ecosystem, diazotrophs must respond to varied environmental conditions to regulate the tremendously taxing nitrogen fixation process. All characterized diazotrophs regulate nitrogenase at the transcriptional level. A smaller set also possesses a fast-acting post-translational regulation system. Although there is little apparent variation in the sequences and structures of nitrogenases, there appear to be almost as many nitrogenase-regulating schemes as there are nitrogen fixing species. Herein are described the paradigms of nitrogenase function, transcriptional control and post-translational regulation, as well as the variations on these schemes, described in various nitrogen-fixing bacteria. Regulation is described on a molecular basis, focusing on the functional and structural characteristics of the proteins responsible for control of nitrogen fixation. PMID- 10801901 TI - Metallothionein expression in animals: a physiological perspective on function. AB - An integration of knowledge concerning regulation of metallothionein expression with research on metallothionein's proposed functions is necessary to delineate how this metalloprotein affects cellular processes, especially zinc metabolism. Metallothionein expression is driven by a number of physiological mediators through several response elements in the metallothionein gene promoter. Cellular accumulation of metallothionein depends on both gene expression and protein degradation. Both depend largely on availability of cellular zinc derived from the dietary zinc supply. Metallothionein expression is related to zinc accumulation in certain organs. Evidence has been produced, which suggests that metallothionein could act in a number of biochemical processes. It may act in zinc trafficking and/or zinc donation to apoproteins, including zinc finger proteins that act in cellular signaling and transcriptional regulation. As a result, metallothionein expression may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. The ability of metallothionein to exchange other metals with zinc in these proteins may explain a role in metal toxicity. Similarly, mobilization of zinc from metallothionein by oxidative stresses may explain its proposed antioxidant function. Apparent good health of metallothionein-deficient mice argues against a critical biological role for metallothionein; however, expression may be critical in times of stress. PMID- 10801902 TI - Kenneth K. Carroll (1923-1998) PMID- 10801903 TI - Wine modifies the effects of alcohol on immune cells of mice. AB - Ethanol may be detrimental to immune cells due to the generation of free radicals during detoxification. If this is true, then alcoholic beverages that contain antioxidants, like red wine, should be protective against immune cell damage. We investigated this by giving mice either a red muscadine wine (Vitis rotundifolia), a cabernet sauvignon (Vitis vinifera), ethanol (all at 6% alcohol) or water in the water bottles as the sole fluid for 8 wk. Plasma antioxidant capacity was measured with alphaalpha-diphenyl-beta-picrylhydrazyl and was more than doubled in the mice that consumed wine compared to control mice that consumed water or ethanol. Cytochrome P450-2E1 levels and glutathione-S transferase activity were modified in such a way as to be interpreted as protective. An immune response was elicited by an intraperitoneal injection of lipopolysaccharide. Later (24 h), natural killer cells and T-lymphocytes derived from the circulation were quantitated in the leukocyte fraction by flow cytometry. Ethanol consumption, as ethanol, significantly suppressed baseline cell numbers relative to the other groups. However, the mice that consumed the same amount of alcohol as wine had baseline cell numbers not different from the water-consuming controls. The lymphocyte response to lipopolysaccharide challenge was inhibited in the mice that consumed ethanol, but was normal in those that consumed the same amount of alcohol in the form of wine. We conclude that there are phytochemicals acting as antioxidants and impacting on the detoxification pathway in the wine that offset the detrimental effects of ethanol on immunity. PMID- 10801904 TI - Docosahexaenoic acid and vitamin E can reduce human monocytic U937 cell apoptosis induced by tumor necrosis factor. AB - The effects of polyunsaturated fatty acids and vitamin E on tumor necrosis factor (TNF)-induced apoptosis of human monocytic U937 cells was explored to assess to what extent these nutrients could attenuate apoptosis. Preincubation of U937 cells with arachidonic acid for 24 h did not affect TNF-induced apoptosis. Eicosapentaenoic acid slightly but significantly reduced the proportion of apoptotic cells only when apoptosis was induced by TNF without cycloheximide (CHI). In contrast, preincubation with docosahexaenoic acid (DHA) greatly (40 approximately 70%) attenuated apoptosis induced by stimulation with either TNF or TNF + CHI for 3 h. The inhibition of apoptosis was accompanied by enrichment of DHA in membrane phospholipids, indicating that DHA probably exerted its inhibitory activity after being incorporated into the phospholipids. Vitamin E also played a role as a partial inhibitor of apoptosis 3 h after TNF addition. This vitamin could further reduce the apoptosis of DHA-treated cells, and such an additive effect was obvious when apoptosis was induced at a low frequency. Longer range stimulation of U937 cells with TNF showed that inhibition of apoptosis by preincubating cells with either DHA or vitamin E was not significant 9 h after TNF addition, but that preincubation with both DHA and vitamin E could reduce the proportion of apoptotic cells even at this time point. Our findings suggested that ingestion of nutrients such as DHA and vitamin E might exert beneficial effects on organ dysfunction associated with various TNF-related diseases. PMID- 10801905 TI - Niacin deficiency in rats increases the severity of ethylnitrosourea-induced anemia and leukopenia. AB - Many chemotherapeutic agents function by damaging the DNA of rapidly dividing cells, leading to side effects in the bone marrow, including anemia and leukopenia during chemotherapy and the development of secondary leukemias in the years following recovery from the original disease. We have created an animal model of alkylation-based chemotherapy, in nontumor-bearing rats, to investigate the effect of niacin deficiency on the side effects of chemotherapy [2 x 2 design, niacin-deficient (ND) vs. pair-fed (PF) control, and ethylnitrosourea (ENU) vs. vehicle control (C)]. Weanling Long-Evans rats were fed ND diet or PF niacin replete diet for 4 wk. ENU or C treatment started after 1 wk of feeding and consisted of 12 doses delivered by gavage, every other day. At 4 wk postweaning, niacin deficiency and ENU treatment ended, the rats were fed a high quality control diet (AIN-93M) and the recovery of blood variables was monitored. ND alone decreased growth rate and caused anemia and neutrophilia. ENU treatment alone caused anemia, lymphopenia, neutropenia and an increase in circulating reticulocytes. In combination, ND and ENU treatment synergistically decreased hematocrit. ND prevented the ENU-induced increase in reticulocyte numbers observed in control rats. ND also increased the severity of ENU-induced lymphopenia. A combination of ND and ENU abolished the neutrophilia caused by ND alone. In summary, ND significantly increased the susceptibility of young Long Evans rats to ENU-induced bone marrow suppression, suggesting that niacin deficient cancer patients may benefit from supplementation. PMID- 10801906 TI - Lymphatic delivery and in vitro pancreatic lipase hydrolysis of glycerol esters of conjugated linoleic acids in rats. AB - We examined the intestinal delivery of conjugated linoleic acids (CLA) given in their triacylglycerol form in the mesenteric lymph of rats. Emulsions containing a mixture of the trilinolein/triester of CLA (9:1) and a tri-[1-(14)C]-linoleyl sn-glycerol tracer were administered by force-feeding. Lymph was collected over two time periods (0-6 and 6-24 h), and the apparent recovery of CLA was determined relative to that of [1-(14)C]-18:2(n-6). A mixture of CLA triester/trilinolein (1:9), trilinolein or CLA-triester was separately subjected to pancreatic lipase hydrolysis in vitro to determine whether the lymphatic recovery of CLA was correlated with the initial step of digestion. Lymphatic recovery of CLA was similar to that of 18:2(n-6) (95.6+/-9.0% of the linoleic acid recovery), and isomer repartition was similar in lymph and in the oil fed, indicating that all the CLA isomers were equally absorbed by the enterocytes. Unexpectedly, the in vitro release of CLA into the absorbable forms (free fatty acids and 2-monoacyl-sn-glycerol) was consistently lower than that of 18:2(n-6). Moreover, the 9c, 11t-isomer of CLA was also released faster into the absorbable forms than its 10t,12c homolog (P = 0.05). We cannot ascribe a distinct cellular accumulation or a difference in the biological effects of different CLA isomers on the ground of a selective intestinal absorbability. Also, the physiological conditions prevailing in vivo in the digestive tract are likely to overcome the relative resistance of CLA ester bonds to pancreatic lipase hydrolysis and allow a lymphatic recovery of CLA similar to that of linoleic acid. PMID- 10801907 TI - Vitamin B-6 deficiency in rats reduces hepatic serine hydroxymethyltransferase and cystathionine beta-synthase activities and rates of in vivo protein turnover, homocysteine remethylation and transsulfuration. AB - Vitamin B-6 deficiency causes mild elevation in plasma homocysteine, but the mechanism has not been clearly established. Serine is a substrate in one-carbon metabolism and in the transsulfuration pathway of homocysteine catabolism, and pyridoxal phosphate (PLP) plays a key role as coenzyme for serine hydroxymethyltransferase (SHMT) and enzymes of transsulfuration. In this study we used [(2)H(3)]serine as a primary tracer to examine the remethylation pathway in adequately nourished and vitamin B-6-deficient rats [7 and 0.1 mg pyridoxine (PN)/kg diet]. [(2)H(3)]Leucine and [1-(13)C]methionine were also used to examine turnover of protein and methionine pools, respectively. All tracers were injected intraperitoneally as a bolus dose, and then rats were killed (n = 4/time point) after 30, 60 and 120 min. Rats fed the low-PN diet had significantly lower growth and plasma and liver PLP concentrations, reduced liver SHMT activity, greater plasma and liver total homocysteine concentration, and reduced liver S adenosylmethionine concentration. Hepatic and whole body protein turnover were reduced in vitamin B-6-deficient rats as evidenced by greater isotopic enrichment of [(2)H(3)]leucine. Hepatic [(2)H(2)]methionine production from [(2)H(3)]serine via cytosolic SHMT and the remethylation pathway was reduced by 80.6% in vitamin B-6 deficiency. The deficiency did not significantly reduce hepatic cystathionine beta-synthase activity, and in vivo hepatic transsulfuration flux shown by production of [(2)H(3)]cysteine from the [(2)H(3)]serine increased over twofold. In contrast, plasma appearance of [(2)H(3)]cysteine was decreased by 89% in vitamin B-6 deficiency. The rate of hepatic homocysteine production shown by the ratio of [1-(13)C]homocysteine/[1-(13)C]methionine areas under enrichment vs. time curves was not affected by vitamin B-6 deficiency. Overall, these results indicate that vitamin B-6 deficiency substantially affects one-carbon metabolism by impairing both methyl group production for homocysteine remethylation and flux through whole-body transsulfuration. PMID- 10801908 TI - Ginger extract consumption reduces plasma cholesterol, inhibits LDL oxidation and attenuates development of atherosclerosis in atherosclerotic, apolipoprotein E deficient mice. AB - Oxidative modification of LDL is thought to play a key role in the pathogenesis of atherosclerosis. Consumption of nutrients rich in phenolic antioxidants has been shown to be associated with attenuation of development of atherosclerosis. This study was undertaken to investigate the ex vivo effect of standardized ginger extract on the development of atherosclerosis in apolipoprotein E deficient (E(0)) mice, in relation to plasma cholesterol levels and the resistance of their LDL to oxidation and aggregation. E(0) mice (n = 60; 6-wk old) were divided into three groups of 20 and fed for 10 wk via their drinking water with the following: group i) placebo (control group), 1.1% alcohol and water (11 mL of alcohol in 1 L of water); group ii) 25 microg of ginger extract/d in 1.1% alcohol and water and group iii) 250 microg of ginger extract/day in 1.1% alcohol and water. Aortic atherosclerotic lesion areas were reduced 44% (P<0.01) in mice that consumed 250 microg of ginger extract/day. Consumption of 250 microg of ginger extract/day resulted in reductions (P<0.01) in plasma triglycerides and cholesterol (by 27 and 29%, respectively), in VLDL (by 36 and 53%, respectively) and in LDL (by 58 and 33%, respectively). These results were associated with a 76% reduction in cellular cholesterol biosynthesis rate in peritoneal macrophages derived from the E(0) mice that consumed the high dose of ginger extract for 10 wk (P<0.01). Furthermore, peritoneal macrophages harvested from E(0) mice after consumption of 25 or 250 microg of ginger extract/day had a lower (P<0.01) capacity to oxidize LDL (by 45 and by 60%, respectively), and to take up and degrade oxidized LDL (by 43 and 47%, respectively). Consumption of 250 microg of ginger extract/day also reduced (P<0.01) the basal level of LDL-associated lipid peroxides by 62%. In parallel, a 33% inhibition (P<0.01) in LDL aggregation (induced by vortexing) was obtained in mice fed ginger extract. We conclude that dietary consumption of ginger extract by E(0) mice significantly attenuates the development of atherosclerotic lesions. This antiatherogenic effect is associated with a significant reduction in plasma and LDL cholesterol levels and a significant reduction in the LDL basal oxidative state, as well as their susceptibility to oxidation and aggregation. PMID- 10801909 TI - High-level dietary vitamin A enhances T-helper type 2 cytokine production and secretory immunoglobulin A response to influenza A virus infection in BALB/c mice. AB - Vitamin A supplementation during acute pneumonia has not improved recovery in most human clinical trials. We hypothesize that high vitamin A intake may decrease the production of T-helper type-1 (Th1) cytokines and thereby inhibit antiviral responses. Such decreases might impair recovery from viral respiratory infections. We thus examined the effect of three interventions on viral pneumonia: 1) a high level vitamin A [250,000 IU/kg diet or 75,000 retinol equivalents (RE)/kg], or 2) control diet (4000 IU/kg diet or 1200 RE/kg) given before and during infection, and 3) initiating the high level diet upon infection to simulate the adjuvant therapy used in clinical trials. No difference was seen among the interventions in severity of disease (weight loss, lung virus titers and survival). However, both the high level diet group and the group in which vitamin A was increased at the time of infection had greater salivary immunoglobulin (Ig)A responses (geometric means, 166 and 105 microg/L, respectively) than did the control group (59 microg/L) (P = 0.0019). In contrast, the serum IgG response was higher in the control group (324+/-158 mg/L) than in the high level group (225+/-95 mg/L) (P = 0.028), although it did not differ from the group in which the diet was changed upon infection (230+/-163 mg/L) (P = 0.084). The production of interferon-gamma (IFN-gamma), a Th1 cytokine, was lower in the high level diet group (median, 0.153 microg/L) compared with the control group (median, 0.839 microg/L) (P = 0.014), whereas the production of interleukin 10 (IL-10), a Th2 cytokine, was higher with the high level diet (median, 0.304 microg/L) than with the control (median, 0.126 microg/L) (P = 0.022). This change in the Th1/Th2 pattern was not sufficient to affect recovery from viral pneumonia but may account for the increased IgA and decreased IgG responses seen with high level dietary vitamin A in this study. These data reinforce the lack of utility of vitamin A in treating acute pneumonia in children and suggest that high dose vitamin A supplements may enhance Th2-mediated immune responses, which are particularly beneficial in the case of extracellular bacterial and parasitic infections and IgA-mediated responses to mucosal infections. PMID- 10801910 TI - Conjugated linoleic acids exhibit a strong fat-to-lean partitioning effect, reduce serum VLDL lipids and redistribute tissue lipids in food-restricted rats. AB - Effects of conjugated linoleic acids (CLA) on a series of metabolic events are expected to depend on the feeding regimen and levels of energy ingested. This study was the first examining the mode of action of CLA on body composition, tissue lipids, lipoproteins and hepatic enzymes in situations of enhanced fat store mobilization. Two groups of male growing Sprague-Dawley rats were fed for 3 wk a diet containing 0 (control group) or 3 g/100 g of a CLA mixture at the expense of sunflower oil, and were then subjected to a weight-loss feeding regimen for another 18 d. Rats fed the CLA-fortified diet gained 11% less weight than the control rats (P<0.05). Rats fed the high CLA diet had less body fat (1.47+/-0.16 vs. 1.07+/-0.09 g/100g, P<0.05) and a higher lean deposition (25.6+/ 0.2 vs. 28.4+/-0.3 g/100 g, P<0.05) than control rats. CLA-fed rats had a 41% lower cholesterol concentration in liver than the control rats (P<0.05). Some differences in glycerophospholipid subclass profile of liver and erythrocyte membrane were observed; the hepatic concentrations of phosphatidylethanolamine (4.76+/-0.46 vs. 6.86+/-0.99 micromol/g, P = 0.07) and phosphatidylcholine (12.9+/-0.5 vs. 15.3+/-1.2 micromol/g, P = 0.09) tended to be greater and the level of phosphatidylcholine in erythrocyte membranes was significantly greater (1.40+/-0.12 vs. 1.83 +/-0.16 micromol/g, P<0.05) in the CLA-treated group than in the control group. The activities of catalase and ornithine decarboxylase in liver did not differ between the groups. Further, CLA-treated rats had significantly lower serum concentrations of VLDL lipids than control rats, whereas concentrations of LDL and HDL lipids were unaffected. The results indicate that a high dose of a CLA mixture is a strong repartitioning agent and a modulator of lipid metabolism under conditions of enhanced fat store mobilization in rats. PMID- 10801911 TI - Insulin-like growth factor-I stimulates renal 1, 25-dihydroxycholecalciferol synthesis in old rats fed a low calcium diet. AB - The adaptive increase in renal proximal tubule 25-hydroxyvitamin D-alpha hydroxylase activity (1-OHase) during dietary calcium restriction is mediated by an increase in parathyroid hormone (PTH) and is inhibited by aging. Recent studies in mature (3-4 mo) rats demonstrated that insulin-like growth factor-I (IGF-I) restored stimulation of renal 1,25-dihydroxycholecalciferol [1,25(OH)(2)D(3)] production by low phosphorus diet (LPD), another major stimulus of 1-OHase. These studies were designed to determine whether IGF-I stimulates 1 OHase during low calcium intake in old rats. Male rats were fed a normal calcium diet (NCD, 6 g Ca/kg diet) or low calcium diet (LCD, 0.2 g Ca/kg diet) for 14 d, and recombinant human IGF-I [rhIGF-I, 1.4 mg/(24h 160 kg body wt)] or vehicle was administrated via miniosmotic pump for 72 h before killing. In 4-mo-old male Sprague-Dawley rats, LCD increased in vitro renal 1-OHase activity in the presence but not in the absence of rhIGF-I. LCD increased in vitro1-OHase activity in young (1-mo-old) but not old (24-mo-old) male Fischer 344 rats. RhIGF I increased 1-OHase activity in 24 mo-old rats fed LCD to levels that were not different from those in 1-mo-old rats fed LCD. The results indicate that the adaptive increase in 1-OHase activity due to a LCD is lost by 4 mo in rats and can be restored by pharmacologic doses of rhIGF-I. PMID- 10801912 TI - Antagonism of arachidonic acid is linked to the antitumorigenic effect of dietary eicosapentaenoic acid in Apc(Min/+) mice. AB - The multiple intestinal neoplasia (Apc(Min/+)) mouse possesses a germline mutation at codon 850 of the adenomatous polyposis coli (Apc) gene resulting in the formation of a nonfunctional truncated gene product. Following a somatic mutation of the remaining wild-type allele, mice spontaneously develop approximately 40-50 tumors throughout the intestinal tract. This mouse model has been used to study intestinal tumorigenesis because this mutation is analogous to the inherited APC mutation in humans with familial adenomatous polyposis (FAP). These individuals characteristically develop numerous adenomas throughout their intestinal tracts. Only a few studies have evaluated the effects of dietary fatty acids on tumorigenesis in this animal model with varying results, and none have linked these effects to alterations in arachidonic acid (AA) metabolism. This study was designed to evaluate the antitumorigenic effect of dietary (n-3) polyunsaturated fatty acids (PUFA) in the Apc(Min/+) mouse model and to determine whether these effects are related to inhibition of AA metabolism. Male Apc(Min/+)mice were fed diets supplemented with eicosapentaenoic acid (EPA), AA or a combination of AA + EPA. Mean tumor number in the EPA group was 68% lower (P<0.05) compared with the control group, whereas AA supplementation did not significantly alter tumor load. The reduction in tumor load coincided with significant reductions in intestinal AA content and levels of prostaglandins. However, supplementing AA to the EPA diet (AA + EPA) abolished the antitumorigenic effect of EPA, increased tissue AA content fourfold and prostaglandin production two- to fourfold. These results indicate that AA is involved in tumorigenesis and suggest that EPA's ability to reduce tumor load in Apc(Min/+) mice is related to reductions in tissue AA content or its metabolism. PMID- 10801913 TI - Vitamin A deficiency injures lung and liver parenchyma and impairs function of rat type II pneumocytes. AB - The purpose of this research was to determine the effects of vitamin A deficiency on liver and lung morphology and type II pneumocyte function. Weanling rats were fed a retinol-adequate (control) or -deficient diet for 6 wk. Average food intakes and body weights were not different between the vitamin A-deficient and adequate rats. Histologic examination revealed that the lungs of vitamin A deficient rats had less collagen in the adventitia of small caliber arteries and arterioles and in the alveolar septa, which appeared thinner than that of controls. Many areas of the lungs of the same rats were also emphysematous (increased size of air spaces distal to the terminal bronchiole, with thinning and partial or total destruction of septal wall). Content of elastin also was lower in the lung parenchyma, as well as in the small arteries and arterioles, but not in the larger ones. Peribronchial collagen was not affected by the deficient diet. Scattered inflammation was observed in most of the vitamin A deficient rats; a mild inflammatory reaction also was seen in one of the controls. Vitamin A-deficient rats also exhibited hepatocyte vacuolization and mild inflammation in the liver, specifically in the periportal tracts. Surfactant synthesis and ornithine decarboxylase activity were significantly lower in type II pneumocytes isolated from vitamin A-deficient rats. In conclusion, our data provide evidence that vitamin A deficiency produces profound morphologic alterations in liver and lung parenchyma and impairs pneumocyte function. PMID- 10801914 TI - Vegetable oils high in phytosterols make erythrocytes less deformable and shorten the life span of stroke-prone spontaneously hypertensive rats. AB - Previous studies have shown that canola oil (CA), compared with soybean oil (SO), shortens the life span of stroke-prone spontaneously hypertensive (SHRSP) rats, a widely used model for hemorrhagic stroke. SHRSP rats are highly sensitive to dietary cholesterol manipulations because a deficiency of membrane cholesterol makes their cell membranes weak and fragile. Phytosterols, abundant in CA but not in SO, can inhibit the absorption of cholesterol and also replace a part of cholesterol in cell membranes. This study was performed to determine whether the high concentration of phytosterols in CA might account for its life-shortening effect on SHRSP rats. Male, 35-d-old SHRSP rats (n = 28/group) were fed semipurified diets containing CA, SO, CA fortified with phytosterols (canola oil + phytosterols, CA + P), SO fortified with phytosterols (soybean oil + phytosterols, SO + P), corn oil (CO), olive oil (OO) or a fat blend that mimicked the fat composition of a representative Canadian diet (Canadian fat mimic, CFM; 10 g/100 g diet). These fats provided 97, 36, 207, 201, 114, 27 and 27 mg phytosterols/100 g diet, respectively. Ten rats from each group were killed after 30-32 d for blood and tissue analyses. The remaining rats (18/group) were used for determination of life span. The life span of SHRSP rats fed the high phytosterol oils (CA, CA + P, SO + P and CO) was significantly (P<0.05) shorter than that of CFM- and SO-fed rats. At 30-32 d, the groups fed the high phytosterol oils had greater levels of phytosterols and significantly (P<0.05) higher ratios of phytosterols/cholesterol in plasma, RBC, liver and kidney, and a significantly (P<0.05) lower RBC membrane deformabilty index than the groups fed oils low in phytosterols (SO, OO and CFM). The mean survival times were correlated with RBC deformability index (r(2) = 0.91, P = 0.0033) and cholesterol concentration (r(2) = 0.94, P = 0.0016), and inversely correlated with RBC phytosterol concentration (r(2) = 0.58, P = 0.0798) and phytosterols/cholesterol (r(2) = 0.65, P = 0.0579), except in the OO group. This study suggests that the high concentration of phytosterols in CA and the addition of phytosterols to other fats make the cell membrane more rigid, which might be a factor contributing to the shortened life span of SHRSP rats. PMID- 10801915 TI - Proliferation and differentiation of stromal-vascular cells in primary culture differ between neonatal pigs consuming maternal or formula milk. AB - Proliferation and differentiation of preadipocytes from 7-d-old pigs consuming maternal or formula milk were examined in primary culture of stromal-vascular (s v) cells derived from subcutaneous adipose tissue. Unsuckled pigs were bottle-fed isoenergetically with colostrum and then sow's milk (SM) or with formula milk alone (F) from birth to 7 d. Isolated cells were exposed to serum-supplemented medium and serum-free medium to determine proliferation and differentiation, respectively. Proliferation estimated between d 3 and 4 of culture was higher (P<0.05) in cells from F than SM pigs. In addition, the number of s-v cells isolated from 1 g of adipose tissue was higher (P<0.01) in F than SM pigs. Variables assessing differentiation were also affected. The percentage of differentiating cells and lipoprotein lipase (LPL) activity were lower (P<0.05) in F than SM pigs, whereas malic enzyme (ME) activity did not differ significantly between the two groups. In conclusion, formula milk increased the number of s-v cells and their capacity for proliferation, whereas the potential for cell differentiation was lower compared with cells from the maternal milk group. Further studies are required to identify the growth and/or nutritional factors that are implicated in the observed differences and to determine whether subsequent development of adipose tissue is affected. PMID- 10801916 TI - Gene expressions of leptin, insulin receptors and lipogenic enzymes are coordinately regulated by insulin and dietary fat in rats. AB - Regulation of the gene expressions of leptin, insulin receptors and lipogenic enzymes was investigated after refeeding a fat-free diet or a 10 g/100 g corn oil diet to food-deprived rats. Plasma glucose and insulin concentrations began to increase 30 min after the feeding and further increased up until 8 h. In these rats, the expression of leptin mRNA in adipose tissue began to increase significantly only 30 min after feeding, and reached a maximum at 8-16 h. However, plasma leptin levels did not increase until 4 h after refeeding, then markedly increased and reached the maximal level after 8 h. The expression of leptin mRNA and plasma leptin concentrations generally were greater in rats fed the corn oil diet compared to those fed the fat-free diet. Insulin receptor mRNA concentrations in the liver and adipose tissue began to decrease 30 min after the refeeding, in contrast to the plasma insulin increase, and continued to decrease until 8 h. The expression of acetyl-CoA carboxylase and fatty acid synthase mRNA began to increase 4-8 h after feeding and reached maximal levels at 16-24 h. Leptin treatment suppressed the expression of lipogenic enzyme mRNA in rats fed the fat-free diet but not in corn oil-fed rats, in which the expression was suppressed by polyunsaturated fatty acids and leptin expression was higher. Thus, we suggest that the glucose and insulin-dependent expressions of leptin, insulin receptors and lipogenic enzymes are coordinately and/or mutually regulated by dietary manipulation. PMID- 10801917 TI - Carotenoid bioavailability in humans from tomatoes processed in different ways determined from the carotenoid response in the triglyceride-rich lipoprotein fraction of plasma after a single consumption and in plasma after four days of consumption. AB - Tomatoes are the main dietary source of lycopene, and the bioavailability of lycopene from tomato paste is higher than that from fresh tomatoes. We investigated systematically the effect of mechanical homogenization and heating on the bioavailability of carotenoids from canned tomatoes. Further, we compared the carotenoid response in triglyceride-rich lipoproteins (TRL) after single consumption with the change in fasting plasma carotenoid concentrations after 4 d of daily consumption. In a split plot design, 17 men and women consumed tomatoes which had received minimal additional heating and 16 others consumed extensively additionally heated tomatoes (1 h at 100 degrees C). These tomatoes were not, mildly or severely homogenized. The tomato products were consumed daily (ca. 22 mg/d lycopene) for 4 d. Eleven participants provided postprandial blood samples on the d 1 and all gave fasting blood samples on d 1 and 4. Homogenization enhanced the lycopene response significantly (P<0.05) both in TRL [mean areas under the curves: 54.9, 72.0 and 88.7 nmol. h/L (SE 11.0) for not, mildly and severely homogenized tomatoes, respectively] and in plasma [mean changes: 0.19, 0.22 and 0.23 micromol/L (SE 0.009), respectively]. Additional heating also tended to enhance the lycopene responses in TRL (P = 0.14) and plasma (P = 0.17). Similar effects to those for lycopene were found for beta-carotene. We conclude that the intactness of the cellular matrix of tomatoes determines the bioavailability of carotenoids and that matrix disruption by mechanical homogenization and/or heat treatment enhances the bioavailability. The carotenoid response in plasma after 4 d intervention can be used to compare the bioavailability of carotenoids from different foods. PMID- 10801918 TI - Fn-type chicory inulin hydrolysate has a prebiotic effect in humans. AB - The partial enzymatic hydrolysis of chicory inulin (GFn; 2 < or =n < or =60) yields an oligofructose preparation that is composed of both GFn-type and Fn-type oligosaccharides (2 < or =n < or =7; 2 < or =m < or =7), where G is glucose, F is fructose, and n is the number of beta(2-->1) bound fructose moieties. Human studies have shown that feeding GFn-type oligomers significantly modifies the composition of the fecal microflora especially by increasing the number of bifidobacteria. The experiments reported here were used to test the hypothesis that the Fn-type molecules have the same property. During a controlled feeding study, 8 volunteers (5 females and 3 males) consumed 8 g/d of an Fn-rich product for up to 5 wk. Fecal samples were collected and analyzed for total anaerobes, bifidobacteria, lactobacilli, bacteroides, coliforms and Clostridium perfringens. Both 2 and 5 wk of oligofructose feeding resulted in a selective increase in bifidobacteria (P<0.01). In addition, a daily intake of 8 g of the Fn-type oligofructose preparation reduced fecal pH and caused little intestinal discomfort. PMID- 10801919 TI - Bioavailabilities of quercetin-3-glucoside and quercetin-4'-glucoside do not differ in humans. AB - The flavonoid quercetin is an antioxidant which occurs in foods mainly as glycosides. The sugar moiety in quercetin glycosides affects their bioavailability in humans. Quercetin-3-rutinoside is an important form of quercetin in foods, but its bioavailability in humans is only 20% of that of quercetin-4'-glucoside. Quercetin-3-rutinoside can be transformed into quercetin 3-glucoside by splitting off a rhamnose molecule. We studied whether this 3 glucoside has the same high bioavailability as the quercetin-4'-glucoside. To that end we fed five healthy men and four healthy women (19-57 y) a single dose of 325 micromol of pure quercetin-3-glucoside and a single dose of 331 micromol of pure quercetin-4'-glucoside and followed the plasma quercetin concentrations. The bioavailability was the same for both quercetin glucosides. The mean peak plasma concentration of quercetin was 5.0+/-1.0 micromol/L (+/-SE) after subjects had ingested quercetin-3-glucoside and 4.5+/-0.7 micromol/L after quercetin-4' glucoside consumption. Peak concentration was reached 37 +/-12 min after ingestion of quercetin-3-glucoside and 27+/-5 min after quercetin-4'-glucoside. Half-life of elimination of quercetin from blood was 18.5+/-0.8 h after ingestion of quercetin-3-glucoside and 17.7+/-0.9 h after quercetin-4'-glucoside. We conclude that quercetin glucosides are rapidly absorbed in humans, irrespective of the position of the glucose moiety. Conversion of quercetin glycosides into glucosides is a promising strategy to enhance bioavailability of quercetin from foods. PMID- 10801920 TI - Culturally appropriate nutrition education improves infant feeding and growth in rural Sichuan, China. AB - Chinese studies indicate that the growth of rural infants and children lags behind that of their urban counterparts after 4 mo of age and that the gap is widening. However, the rural areas are home to >85% of China's 300 million children. Clearly, culturally appropriate rural complementary feeding interventions are needed to close the growth and health gaps. After a 1990 survey of infants in rural Sichuan confirmed that poor infant feeding practices rather than inadequate household food resources were responsible for the growth faltering, a year-long community-based pilot nutrition education intervention (n congruent with 250 infants each in Education and Control groups) was undertaken in four townships. The goal was to improve infant growth by improving infant feeding practices. Features of the intervention included the training and mobilizing of village nutrition educators who made monthly growth monitoring and complementary feeding counseling visits to all pregnant women and families with infants born during the intervention in the study villages. After 1 y, the Education group mothers showed significantly higher nutrition knowledge and better reported infant feeding practices than their Control group counterparts. Also, the Education group infants were significantly heavier and longer, but only at 12 mo (weight-for-age -1.17 vs. -1.93; P = 0.004; height-for-age -1.32 vs. 1.96; P = 0.022), had higher breast-feeding rates overall (83% vs. 75%; P = 0.034) and lower anemia rates (22% vs. 32%; P = 0.008) than the Control group infants. We conclude that these methods have potential for adaptation and development to other rural areas in the county, province and nation. PMID- 10801921 TI - Iron deficiency is a more important cause of anemia than hemoglobinopathies in Kuwaiti adolescent girls. AB - Anemia is the most prevalent nutritional problem worldwide, due mainly to iron deficiency. Studies of anemia are less common in adolescents than in women and children. We examined anemia prevalence in adolescent Kuwaiti schoolgirls, and its association with hemoglobinopathies as well as the most common environmental cause, Fe deficiency. A cross-sectional sample of 1051 healthy adolescent schoolgirls was studied. Sample size was based on WHO criteria. Anemia, Fe deficiency and hemoglobin (Hb) variations were studied by Hb concentration, erythrocyte protoporphyrin (EP) and an HPLC quantitation, respectively. Of the subjects sampled, 30% were anemic. Mildly elevated EP values were found in 68%. Girls with high EP levels were more likely (P < 0.001) to be anemic than girls with normal EP. Up to 25% of the girls may have had Fe deficiency anemia. Hemoglobinopathies were neither prevalent nor significantly associated with anemia. These data indicate that environmental factors play a significant role in anemia among healthy, well-to-do Kuwaiti adolescent girls. PMID- 10801922 TI - Pork fat and chicken fat similarly affect plasma lipoprotein metabolism in cynomolgus monkeys fed diets with adequate levels of linoleic acid. AB - The effects on plasma lipoprotein metabolism of replacing pork fat (PF) with chicken fat (CF) (formulated as part of currently recommended prudent diets) was evaluated in 10 male cynomolgus monkeys. Monkeys were rotated through three dietary periods, (each of 10-wk duration), during which total cholesterol (TC), triacylglycerol (TG) and HDL-cholesterol (HDL-C) were measured (7, 8 and 9 wk) and in vivo lipoprotein metabolism evaluated (after 9 wk). Initially, all monkeys were fed a high-fat, high-cholesterol reference diet [38% of energy (en) from fat, 18%en saturated fatty acids (SFA), 10%en monounsaturated fatty acids (MUFA), 10%en polyunsaturated fatty acids (PUFA), 0.045 mg cholesterol/kJ diet]. Subsequently, monkeys were rotated through two test diets (30%en fat, SFA/MUFA/PUFA 1:1:1, 0.004-0.005 mg cholesterol/kJ diet), in which 80% of the fat was either PF or CF, with the remaining 20% derived from high-linoleic safflower oil. There was no significant difference between the two test diets for TG, TC, nonHDL-C, HDL-C or the ratio of TC/HDL-C. Lipoprotein composition, LDL apolipoprotein B pool size, fractional catabolic rate and transport rate were also not significantly different when monkeys consumed the two test diets. These data suggest that when incorporated into diets following current guidelines and containing adequate PUFA ( approximately 7-9%en), PF and CF similarly affect plasma lipids. PMID- 10801923 TI - The human gut bacteria Bacteroides thetaiotaomicron and Fusobacterium varium produce putrescine and spermidine in cecum of pectin-fed gnotobiotic rats. AB - Pectin is a soluble indigestible polysaccharide that stimulates cecal polyamine formation in rats. Bacteroides and fusobacteria, two numerically dominant bacterial population groups in the large intestine, were found to synthesize in vitro high amounts of spermidine and putrescine. The purpose of this study was to elucidate the effect of pectin on the polyamine production by defined bacterial species in vivo. Germfree male Wistar rats (n = 18) were randomly assigned to one of three treatments: (i) monoassociation with Bacteroides thetaiotaomicron + fiber-free diet; (ii) diassociation with B. thetaiotaomicron + Fusobacterium varium + fiber-free diet or (iii) diassociation with B. thetaiotaomicron + F. varium + fiber-free diet + 10% pectin. The cecal contents of monoassociated rats fed fiber-free diet contained large amounts (1.51+/-0.21 micromol/dry total cecum content) of spermidine which was the major polyamine. The cecum of diassociated rats fed the fiber-free diet contained even higher concentrations of spermidine (2.53+/-0.21 micromol/dry total cecum content) and also putrescine, which was now the dominant polyamine (putrescine 0.32+/-0.28 vs. 3.01+/-0.28 micromol/dry total cecum content; monoassociation vs. diassociation). Pectin consumption by diassociated rats led to an additional increase in the cecal concentrations of all polyamines: putrescine, spermidine and spermine were 40, 37 and 100%, respectively, higher in the diassociated rats consuming the pectin diet than in those consuming the pectin-free diet. Since the microbial counts in the cecum did not differ in the diassociated treatment groups, the elevated concentrations of polyamines observed in the pectin group must have been due to stimulated bacterial polyamine synthesis. The decline of individual polyamines from cecum to feces detected at the end of the study in all treatment groups and the high microbial counts in the cecum and in feces suggest that bacterial polyamines are absorbed in cecum and colon. Pectin stimulates intestinal microbes to synthesize large amounts of polyamines which may be utilized by the host. PMID- 10801924 TI - Dietary stearic acid reduces cholesterol absorption and increases endogenous cholesterol excretion in hamsters fed cereal-based diets. AB - The observation that dietary stearic acid does not raise plasma cholesterol concentration is well documented, although the regulating mechanisms are not completely understood. Therefore, we examined the effect of dietary stearic acid on cholesterol absorption and sterol balance using male Syrian hamsters fed modified NIH-07 cereal-based diets selectively enriched in palmitic acid (16:0), stearic acid (18:0), trans fatty acid (18:1t), cis oleic acid (18:1c) or linoleic acid (18:2). All diets contained 17 g/100 g total fat and 0.05 g/100 g cholesterol; the five fat blends were enriched 30% with the fatty acid of interest above a constant fatty acid background. Cholesterol absorption efficiency was 50-55% in all treatment groups except for the 18:0 group, in which cholesterol absorption was significantly reduced to 21%. Plasma total cholesterol concentration was significantly lower in the 18:0 group compared to the 16:0 group. Fecal neutral steroid excretion was significantly greater in hamsters fed the high 18:0 diet compared to the other treatment groups. After accounting for unabsorbed dietary cholesterol, endogenous cholesterol excretion was about 100% higher in the 18:0 group. Consequently, the calculated rate of whole body cholesterol synthesis was significantly increased by dietary 18:0. Bile acid excretion accounted for only 12-20% of total sterol output by the hamsters in this study. Thus, the data suggest that reduced plasma cholesterol concentration in hamsters fed high 18:0 diets may be influenced by reduced cholesterol absorption and increased excretion of endogenous cholesterol. PMID- 10801925 TI - Glutathione turnover is increased during the acute phase of sepsis in rats. AB - Glutathione metabolism during infection has been poorly documented. Glutathione concentrations and synthesis rates were studied in infected rats (2 d after infection) and in pair-fed controls. Glutathione synthesis rates were determined in liver, spleen, lung, small and large intestine, skeletal muscle, heart and blood by a 4-h or 6-h (15)N cysteine infusion. The activities of four hepatic enzymes involved in glutathione metabolism were also determined. Glutathione synthesis rates were significantly greater in liver (+465%), spleen (+388%), large intestine (+109%), lung (+100%), muscle (+91%) and heart (+80%) of infected rats compared with pair-fed controls. Glutathione concentrations were also greater in these tissues but were unaffected in small intestine and lower in blood. In keeping with the stimulation of liver glutathione synthesis, the activities of liver gamma-glutamyl-cysteine synthetase and glutathione reductase were significantly greater in liver of infected rats than of pair-fed rats. From the present study, we estimate that glutathione synthesis accounts for at least 40% of the enhanced cysteine utilization during infection. This increased utilization may be the primary cause of an enhanced cysteine requirement in infection. PMID- 10801926 TI - Accumulation of advanced glycation endproducts in aging male Fischer 344 rats during long-term feeding of various dietary carbohydrates. AB - The observation of accelerated collagen glycation in association with enhanced progression of many age-associated diseases in hyperglycemic subjects has led researchers to propose a role of glycation in the aging process. Although short term studies in healthy animals suggest that feeding a diet high in fructose may increase serum glucose concentrations and increase glycemic stress, the effects of a long-term feeding, i.e., life span, are unknown. This study was designed to evaluate the long-term effects of dietary carbohydrates on serum and tissue markers of glycemic stress. Three-month-old male Fischer 344 rats were given free access to or restricted to 60% caloric intake of one of five isocaloric diets that contained as their carbohydrate source either cornstarch, glucose, sucrose, fructose or equimolar amounts of fructose and glucose. Rats were killed at 9-, 18 or 26-mo of age. Glycated hemoglobin, serum glucose and fructosamine levels were measured as markers of serum glycemic stress. Collagen-associated fluorescence and pentosidine concentrations were measured in skin, aortic, tracheal and tail tendon collagen as markers of advanced glycation endproducts (AGE). The source of dietary carbohydrate had little effect on markers of glycemic stress and the accumulation of AGE. Restricting the amount of calories consumed resulted in lower serum glucose concentrations, glycated hemoglobin levels and pentosidine concentrations in tail tendon collagen. Our data suggest that the rate of collagen glycation is tissue-specific. These results suggest that long-term feeding of specific dietary carbohydrates does not alter serum glucose concentrations or the rate of collagen glycation. Rather, age-related accumulation of AGE is more closely related to caloric intake. PMID- 10801927 TI - Tissue distribution of lycopene in ferrets and rats after lycopene supplementation. AB - To determine lycopene uptake and tissue distribution in ferrets (Mustela putorius furo) and F344 rats, we supplemented orally 4.6 mg/(kg body wt.d) lycopene in a tomato oleoresin-corn oil mixture (experimental groups). After 9 wk of supplementation, the animals were killed and blood and organs were collected. Plasma and tissue carotenoids were extracted and measured using HPLC. Mean concentrations of lycopene (nmol/kg wet tissue) in saponified tissues of ferrets were as follows: liver 933, intestine 73, prostate 12.7 and stomach 9.3. Levels of lycopene (nmol/kg wet tissue) in saponified tissue of rats were as follows: liver 14213, intestine 3125, stomach 78.6, prostate 24 and testis 3.9. When these organs were extracted without saponification, the lycopene levels were lower, except for rat testis. All-trans-lycopene was the predominant isomer found in tomato oleoresin and in the majority of rat tissues, whereas cis-lycopenes were predominant in rat prostate and plasma. This pattern was reversed in ferrets. The results show the following: 1) lycopene from tomato oleoresin is absorbed and stored primarily in the liver of both animals; 2) saponification generally improves the extraction of lycopene from most tissues of both animals; 3) cis lycopene and all-trans-lycopene are the predominant isomers in ferret and rat tissues, respectively; and 4) rats absorb lycopene more effectively than ferrets. PMID- 10801928 TI - Glucose-based oligosaccharides exhibit different in vitro fermentation patterns and affect in vivo apparent nutrient digestibility and microbial populations in dogs. AB - To evaluate the potential of indigestible oligosaccharides (OS) to serve as "dietary fiber-like" ingredients, it is necessary to determine their extent of indigestibility. In vitro fermentation characteristics of two novel OS, alpha glucooligosaccharides (GOS) and a maltodextrin-like OS (MD), were compared to those of fructooligosaccharides (FOS), gum arabic (GA), guar gum (GG) and guar hydrolysate (GH). Total short-chain fatty acid (SCFA) production (micromol/g dry matter) as a result of MD fermentation was higher initially compared with GA (P<0.01), but GA was more extensively fermented at 24 h (P<0.01). Total SCFA production for GOS was similar to that for FOS, GG, GH and GA. In the second experiment, GOS and MD were added at 6% to an enteral formula control diet (Control) and fed to ileal-cannulated dogs in a 3x3 replicated Latin-square design. Ileal digestibility of glucose was lower (P<0.05) and carbohydrate (CHO) numerically lower (P = 0.08) for both GOS and MD compared with the Control. Total tract digestibility of CHO and glucose was lower only for MD (P<0.01) compared with the Control. Total fecal weights were higher (P<0.01) for both GOS and MD treatments. Fecal concentration of bifidobacteria was numerically increased by GOS and MD supplementation (P = 0.13 and 0.23, respectively). Thus, GOS and MD are indigestible yet fermentable OS, and may act as "dietary fiber-like" ingredients. PMID- 10801929 TI - High chromium yeast supplementation improves glucose tolerance in pigs by decreasing hepatic extraction of insulin. AB - Twenty Landrace x Yorkshire cross pigs (body wt, 47.9+/-2.9 kg) were used to evaluate effects of dietary high chromium (Cr) yeast supplementation on plasma kinetics of glucose, insulin and C-peptide. Pigs were provided free access to either a control diet (C) containing 204 microg Cr/kg or a diet supplemented with an additional 200 microg Cr/kg as high Cr yeast (CR) for between 23 and 30 d. After overnight food deprivation, dextrose (500 g/L) was infused through a jugular vein catheter at a dose of 0.5 g glucose/kg body weight with an infusion rate of 10 g glucose/min within 6 min. High Cr yeast supplementation did not affect body weight gain or food intake. There were no differences in fasting plasma concentrations of either glucose or C-peptide, although basal plasma concentration of insulin tended to be higher in pigs fed CR (P<0.10). Plasma glucose concentrations were lower (P<0.01) at postinfusion times 5, 10, 15 and 20 min in pigs fed CR. Plasma insulin concentrations in pigs fed CR were higher (P<0.05) at 2 and 0 min before the completion of dextrose infusion. However, the increase in plasma insulin concentrations was not accompanied by a comparable elevation in plasma C-peptide concentrations. The 30-min (postinfusion) area of plasma glucose concentrations tended to be lower (P<0.10) in pigs fed CR, but there were no differences in 30-min areas of either plasma insulin or plasma C peptide concentrations between treatments. Plasma clearance rates of glucose, insulin and C-peptide were higher and their half-lives shorter (P<0.05) in pigs fed CR. In conclusion, dietary high Cr yeast supplementation improved glucose tolerance, possibly through a decrease in hepatic extraction of insulin. PMID- 10801930 TI - Regulation of hepatic vitamin A storage in a rat model of controlled vitamin A status during aging. AB - It is currently unknown whether the capacity of the liver to esterify and store vitamin A (VA) changes as a function of long-term VA intake or age. The objective of this study was to investigate whether age and/or VA status are factors for the hepatic expression of cellular retinol-binding protein (CRBP), the esterification of retinol by lecithin:retinol acyltransferase (LRAT) and the accumulation of VA and lipids in liver. Two factors, VA intake and age, were studied in a 3x3 design. Diets denoted as VA-marginal, control and supplemented contained 0.35, 4 and 25 mg retinol equivalents/kg diet, respectively; male Lewis rats were fed these diets from weaning until the ages of 2-3 mo (young), 8-10 mo (middle-aged) and 18-20 mo (old) (n = 6/group. Liver CRBP mRNA differed (two-way ANOVA) with dietary VA (P<0.0001) and age (P<0.05). Hepatic LRAT activity increased with dietary VA (P<0.0001). Age was not a factor (P = 0.47) although there was an interaction of age and dietary VA (P<0.0001). Hepatic LRAT activity was correlated (r = 0.633, P<0.0001) with plasma retinol at physiologic concentrations. In VA-supplemented rats of all ages, the plasma molar ratio of total retinol:retinol-binding protein (RBP) exceeded 1, and liver VA and total lipid concentrations were elevated. However, tests of liver function had previously been shown to be within normal values. Thus, the capacity of the liver for retinol esterification by LRAT was not diminished by age or the accumulation of VA and other lipids. We conclude the following: 1) hepatic LRAT activity is regulated across a broad, physiologic range of dietary VA; 2) LRAT activity is regulated throughout life; and 3) the capacity for hepatic VA storage is high throughout life. PMID- 10801931 TI - Dietary copper influences reproduction in cats. AB - The objective of this study was to determine the copper requirement of female cats (queens) for gestation. Cuproenzyme activities were evaluated to identify a noninvasive indicator of copper status. This study used a depletion-repletion model. Specific pathogen-free queens (n = 28) were adapted to a purified diet; after consuming a copper-depletion diet (0.8 mg Cu/kg diet) for 4 mo, they were randomly allocated to three dietary treatment groups receiving copper sulfate at 4.0, 5.8 or 10.8 mg Cu/kg diet. Four queens underwent liver biopsies at two time points during the study. Plasma samples were analyzed for copper concentrations, extracellular superoxide dismutase, ceruloplasmin and diamine oxidase activities. Only liver copper concentrations were responsive to dietary copper intake. The dietary concentration of copper had a significant effect on the time taken for queens to conceive (P = 0.04). There was a negative linear relationship between dietary copper (x = Cu mg/kg diet) and the mean time (y = days) for queens to conceive (y = 43.38-2.87x; R(2) = 0.97). The current NRC recommendation of 5 mg/kg diet copper for cats appears marginal for optimal reproduction. PMID- 10801932 TI - Kinetic analysis shows that iron deficiency decreases liver vitamin A mobilization in rats. AB - In view of evidence that nutritional status of iron and vitamin A may affect the other nutrient's metabolism, we used model-based compartmental analysis to examine effects of iron deficiency on whole-body vitamin A dynamics in rats. Weanling male Sprague-Dawley rats were fed the AIN93G diet with 2.5 nmol retinyl palmitate/g and either 45 [control (CN)] or 4 microg/g Fe [iron-deficient (ID)] for 8 wk. ID rats consumed food ad libitum; CN rats were food-restricted so that their body weights were the same as ID rats. Two rats/group were killed; liver vitamin A was determined and used for vitamin A balance calculations. [(3)H]Retinol-labeled plasma was administered intravenously to remaining rats, and 27 serial blood samples were collected for 7 wk. At killing, plasma vitamin A was 0.52+/-0.12 (ID, n = 5) vs. 1.34+/-0.12 micromol/L (CN, n = 6; P<0.001), and liver vitamin A was 809+/-94 (ID) vs. 112+/-24 nmol (CN, P<0.001). Plasma tracer data were fit to a three- or four-compartment model using the Simulation, Analysis and Modeling computer program and kinetic parameters were calculated. Vitamin A transfer rate between the retinyl ester storage pool [14+/-3 (ID) vs. 24+/-4 nmol/d (CN), P<0.05] and plasma was lower in ID rats. Vitamin A remained longer in the body [44+/-11 (ID) vs. 22+/-3 d (CN), P<0.05]. Adjusted mean disposal rate was lower in ID (10.0) than CN rats (19.9 nmol/d), as was estimated vitamin A absorption efficiency [58% (ID) vs. 76% (CN)]. Our results suggest that iron deficiency inhibits mobilization of vitamin A stores and may decrease the absorption and irreversible utilization of vitamin A. PMID- 10801933 TI - Intracellular accumulation of ascorbic acid is inhibited by flavonoids via blocking of dehydroascorbic acid and ascorbic acid uptakes in HL-60, U937 and Jurkat cells. AB - In HL-60, U937 and Jurkat cells, the intracellular accumulation of ascorbic acid occurred via uptakes of both dehydroascorbic acid (an oxidized metabolite of ascorbic acid) and ascorbic acid (vitamin C). Dehydroascorbic acid and ascorbic acid were transported into cells by sodium-independent glucose transporters (GLUT 1 and GLUT 3) and sodium-dependent ascorbic acid transporters, respectively. Flavonoids inhibited the intracellular accumulation of ascorbic acid by blocking dehydroascorbic acid and ascorbic acid uptakes in the transformed cells. At flavonoid concentrations of 10-70 micromol/L, approximately 50% of dehydroascorbic acid uptake was inhibited in the cells. In Jurkat cells, two potent flavonoids (myricetin and quercetin) competitively inhibited dehydroascorbic acid uptake, and K(i) values were approximately 14 and 15 micromol/L, respectively. Because GLUT 1 and GLUT 3 transport dehydroascorbic acid, the inhibition of dehydroascorbic acid uptake by flavonoids was investigated by using Chinese hamster ovary cells overexpressing rat GLUT 1 or human GLUT 3. Myricetin at concentrations of 22 and 18 micromol/L, respectively, inhibited half of dehydroascorbic acid uptake in the cells overexpressing GLUT 1 and GLUT 3. Myricetin also inhibited ascorbic acid uptake; inhibition was noncompetitive with K(i) = 14 micromol/L in Jurkat cells. These data indicate that flavonoids inhibit both ascorbic acid and dehydroascorbic acid uptake but do so by different mechanisms. These data may contribute to new understanding of the biological effect of flavonoids on the intracellular accumulation of ascorbic acid in human cells. PMID- 10801934 TI - Canthaxanthin supplementation alters antioxidant enzymes and iron concentration in liver of Balb/c mice. AB - The 4,4'-diketo-beta-carotene, canthaxanthin, alters tocopherol status when fed to Balb/c mice, suggesting an involvement of carotenoids in the modulation of oxidative stress in vivo. We investigated further the modifications induced by an oral administration of canthaxanthin on lipid peroxidation, antioxidant enzymes and iron status in liver of Balb/c mice. Female 6-wk-old Balb/c mice were randomly divided into two groups (n = 10/group). The control group (C) received olive oil alone (vehicle) and the canthaxanthin-treated group (Cx) received canthaxanthin at a dose of 14 microg/(g body wt.d). The 15-d canthaxanthin treatment resulted in carotenoid incorporation but did not modify lipid peroxidation as measured by endogenous production of malondialdehyde (MDA). However, glutathione peroxidase activity was 35% lower (P<0.01) and catalase (59%, P<0.005) and manganese superoxide dismutase (MnSOD) (28%, P<0.05) activities were higher in canthaxanthin-treated mice than in controls. Moreover, carotenoid feeding caused a significant (P<0.05) overexpression of the MnSOD gene; mRNA levels of the enzyme were greater in treated mice than in controls. Concomitantly, a 27% (P<0.05) greater iron concentration was found in liver from canthaxanthin-treated mice compared with controls. These findings support the hypothesis that canthaxanthin alters the protective ability of tissues against oxidative stress in vivo. PMID- 10801935 TI - Long-term consumption of red wine does not modify intestinal absorption or status of zinc and copper in rats. AB - Red wines contain many components such as polyphenols and ethanol that may influence mineral absorption. We report on studies in a rat model that were designed to investigate the extent to which short- and long-term intake of red wine or ethanol may influence (67)Zn and (65)Cu absorption in rats. Rats (n = 96) were divided into three groups, a control group that received demineralized water, a group that received red wine diluted with water (v/v) and an ethanol group that received 6% ethanol. Half of each group was used for the short-term study; the others were used for the long-term study. After 3 d (short-term study) or 28 d (long-term study) of beverage consumption, the rats were gavaged with 2 mL of solution containing 2027 nmol (67)Zn and 902 nmol (65)Cu. Subsequently, 3-d urinary and fecal collections were performed and analyzed for total and isotopic Zn and Cu. In the long-term study, blood, tibia and liver were also sampled for mineral status assessment. Neither short- nor long-term intake of red wine altered (67)Zn or (65)Cu absorption. In contrast, long-term (but not short-term) ethanol consumption significantly increased both (67)Zn and (65)Cu absorption compared with the control and red wine groups. The long-term consumption of ethanol or red wine did not affect blood or tissue Zn or Cu levels. In conclusion, short- or long-term consumption of red wine did not have a negative effect on intestinal absorption or tissue levels of zinc and Cu in rats. PMID- 10801936 TI - Dietary oligofructose lessens hepatic steatosis, but does not prevent hypertriglyceridemia in obese zucker rats. AB - We studied the influence of oligofructose (OFS), a nondigestible fructan, on lipid metabolism in obese fa/fa Zucker rats. The addition of 10 g/100 g OFS to the diet slowed the increase in body weight without modifying serum triglycerides or glucose concentrations after 7 wk of treatment. However, an oral load of 2 g glucose and 5 g corn oil/kg body weight increased triglyceridemia more in OFS-fed rats than in control rats. After 10 wk, OFS decreased the hepatic concentration of triglycerides 57% relative to controls. The less severe steatosis was confirmed by histologic analysis. Among the key enzymes involved in fatty acid synthesis and esterification, only malic enzyme activity was significantly lower in OFS-fed rats than in controls. The epididymal fat mass was significantly lower in OFS-fed rats. In conclusion, dietary enrichment with OFS can counteract both the fat mass development and the hepatic steatosis that occur in obese Zucker rats. Future studies will be designed to clarify in obese animals the influence of dietary OFS on postprandial triglyceridemia, which is an important variable associated with the development of atherosclerosis in humans, and to analyze the biochemical mechanism underlying the "hepatoprotective" effect of OFS. PMID- 10801937 TI - Time course and pattern of compensatory ingestive behavioral adjustments to lysine deficiency in rats. AB - We and others have demonstrated that rats deficient in an essential amino acid (EAA) will consume sufficient quantities of the lacking nutrient to produce repletion when it is made available in solution. In the current series of experiments, we made rats deficient in lysine (LYS) by limiting the level of this EAA in the diet. We then examined licking behavior during approximately 23-h two bottle intake tests over 4 consecutive days. In three separate experiments, rats were presented with the following: 1) 0.1 mol/L LYS and water, 2) 0.2 mol/L threonine (THR) and water and 3) 0.1 mol/L LYS and 0.2 mol/L THR. Lysine deficient (LYS-DEF) rats drink significantly more LYS than did nondepleted controls (CON) when this amino acid was available. Meal pattern analysis revealed that the enhanced intake of LYS occurred as a function of a greater number of ingestive bouts, not changes in bout size. A cumulative analysis of LYS intake between CON and LYS-DEF rats revealed that a potentiation of intake developed within 30 min of sampling the solution when LYS and water were available and within 90 min when LYS and THR were the contrasting choices. In conclusion, increased LYS intake in the deficient rats occurs relatively rapidly and appears to be at least somewhat specific. Moreover, LYS deficiency does not seem to enhance the palatability of the limiting amino acid as judged by behaviors such as lick rate and bout size. Instead, LYS-DEF rats relieve the deficiency by increasing the number of drinking episodes initiated. PMID- 10801938 TI - Caco-2 cells can be used to assess human iron bioavailability from a semipurified meal. AB - A Caco-2 cell model with extrinsic radioiron was used to evaluate the effect of dietary factors on nonheme iron bioavailability from a semipurified meal. Study 1 was conducted to evaluate the effect of enhancers (ascorbic acid) and inhibitors (bran, phytate and tea) on iron bioavailability when added to semipurified meal containing egg albumen as a protein source. The effect of various proteins [bovine serum albumin (BSA), casein, beef and soy] on iron bioavailability was evaluated in Study 2 by substituting the above protein sources for egg albumen. Protein solubilization following in vitro digestion for individual test meals was not significantly different from the control. On the other hand, nonheme iron solubilization (0.8+/-0.0 to 5.9+/-0.3 vs. 4.9+/-0.8 mg/L) varied significantly. The total iron uptake for each meal was calculated based on the percentage of radioiron taken up and transported by Caco-2 cells and the amount of nonheme iron present in uptake solutions. Iron uptake ratios represent test/control values. With the exception of BSA and ascorbic acid, the effect of dietary factors was similar to that found in humans reported in the literature. A significant correlation (r = 0.97; P<0.0001) was found between the published human absorption data and the iron uptake by the Caco-2 cells. The results of our study indicate the usefulness of Caco-2 cells in assessing human iron absorption and the feasibility of this cell model in studying iron bioavailability from various food combinations, otherwise not easily performed in humans. PMID- 10801939 TI - Length of exclusive breastfeeding: linking biology and scientific evidence to a public health recommendation. PMID- 10801940 TI - Introduction PMID- 10801941 TI - Human zinc deficiency. AB - The objective of this paper is to provide a current overview of the significance of zinc in human nutrition. To achieve this, the following issues are addressed: (1) the biochemistry and biology of zinc in the context of their relevance to zinc in human nutrition and to our understanding of the complexity and practical importance of human zinc deficiency; (2) the history of our understanding of human zinc deficiency with an emphasis both on its brevity and on notable recent progress; (3) the clinical spectrum of severe zinc deficiency; (4) the lack of ideal biomarkers for milder zinc deficiency states, with the consequent dependence on randomized, placebo-controlled intervention studies to ascertain their prevalence and clinical consequences, including growth delay, diarrhea, pneumonia, other infections, disturbed neuropsychological performance and abnormalities of fetal development; (5) the public health significance of human zinc deficiency in the developing world; (6) reasons for concern and unanswered questions about zinc nutriture in the United States; (7) the need for better understanding of human zinc metabolism and homeostasis (including its limitations) at a molecular, cellular, organ-system and whole body level and of factors that affect zinc bioavailability; and (8) potential strategies for the prevention and management of human zinc deficiency. This review concludes with an emphasis on the immediate need for expanded research in directions that have become increasingly well demarcated and impelling as a result of recent progress, which is summarized in this overview. PMID- 10801942 TI - Assessment of marginal zinc status in humans. AB - The assessment of marginal zinc status is problematic. Currently, there is no universally accepted single measure to assess zinc status in humans. The development of a reliable measure of marginal or moderate zinc status would be useful for a variety of purposes. For example, a simple, yet sensitive and accurate measure of zinc nutritional status is critically needed to further our limited understanding of the possible associations between zinc status and the risk of developing various chronic diseases and in predicting favorable health outcomes in patient populations. A convenient and reliable zinc assessment tool is needed to identify subpopulations who are at a risk of zinc deficiency and as an objective guidepost to determine the need for initiation of zinc supplementation or zinc fortification of the food supply, as well in the refinement of recommendations of dietary zinc allowances. In frank zinc deficiency, clinical signs and static measures of zinc concentrations in a variety of readily available tissues, such as plasma, various blood cell types and hair, may uniformly confirm the presence of depleted body zinc stores. However, in general, the relative insensitivity or imprecision of these measurements has resulted in general disappointment in their use to assess marginal zinc status. Therefore, the search continues to find a useful and reliable marker of marginal zinc deficiency. In an attempt to speculate on possible future developments in the zinc status assessment field, a number of new and potentially promising approaches to this problem are highlighted. PMID- 10801943 TI - Kinetics of zinc metabolism: variation with diet, genetics and disease. AB - Kinetic studies are used to investigate metabolic processes. By adding an isotope to a system and measuring its movement in the system over time, pool sizes and transport rates can be determined by mathematically modeling the data. This approach enables rate differences to be determined in conditions that have been modified by diet, environment, genetics or disease. Kinetic studies in humans have shown that there are multiple pools of zinc that turnover from minutes to years and that processes, including zinc absorption and excretion, are regulated to maintain tissue levels when zinc intake varies. Animal studies allow for greater understanding of kinetics because more tissues can be sampled and environmental and genetic factors can be controlled. Kinetic studies in animals will provide information on the overexpression or the deletion of genes coding for specific proteins involved in zinc transport and metabolism. The advances that have been made in our understanding of the role of zinc in metabolism have been aided by the development of techniques for measuring isotopes in biological materials. In the future, the kinetics of zinc bound to different compounds will be measured. Modeling will enable this information, at the molecular level, to be integrated with knowledge of zinc metabolism at the cellular, organ and whole body level. To understand more fully the role of zinc in human health, kinetic studies are needed in healthy and disease states to identify differences in metabolic processes. This knowledge can be used as a basis for dietary and therapeutic recommendations. PMID- 10801944 TI - Zinc homeostasis in humans. AB - Maintaining a constant state of cellular zinc nutrition, or homeostasis, is essential for normal function. In animals and humans, adjustments in zinc absorption and endogenous intestinal excretion are the primary means of maintaining zinc homeostasis. The adjustments in gastrointestinal zinc absorption and endogenous excretion are synergistic. Shifts in endogenous excretion appear to occur quickly with changes in intake just above or below optimal intake. The absorption of zinc responds more slowly, but it has the capacity to cope with large fluctuations in intake. With extremely low zinc intakes or with prolonged marginal intakes, secondary homeostatic adjustments may augment the gastrointestinal changes. These secondary adjustments include changes in urinary zinc excretion, a shift in plasma zinc turnover rates and, possibly, an avid retention of zinc released from selected tissues, such as bone, in other tissues to maintain function. PMID- 10801945 TI - Zinc intake of the U.S. population: findings from the third National Health and Nutrition Examination Survey, 1988-1994. AB - National survey data for 29,103 examinees in the third National Health and Nutrition Examination Survey were used to estimate mean and percentile distributions of dietary and total zinc intakes based on 24-h dietary recalls and vitamin/supplement use. Mean daily total intakes ranged from 5.5 mg in non-breast feeding infants to 13 mg in adults and were higher in adolescent and adult males than in females (P <0.01). Mean total zinc intakes (22 mg) were approximately 10 mg higher in pregnant and lactating females than in nonpregnant, nonlactating females of the same age. Mean total zinc intakes were 0. 7 mg higher in adolescents (11.1 mg) and 2.5-3.5 mg higher in adults (13 mg) compared with mean dietary intakes, indicating the average contribution of supplements to total zinc intake. Mean total zinc intakes were significantly higher in non-Hispanic whites than in non-Hispanic blacks (P<0.01) and Mexican Americans (P<0.01) for men and women aged 51-70 y and > or =71 y due to higher zinc supplement use. The prevalence of zinc-containing supplements use ranged from 0. 1% in infants to 20.5% in adults. "Adequate" zinc intake in this survey population was 55.6% based on total intakes of >77% of the 1989 recommended dietary allowance. Young children aged 1-3 y, adolescent females aged 12-19 y and persons aged > or =71 y were at the greatest risk of inadequate zinc intakes. PMID- 10801946 TI - Overview of zinc absorption and excretion in the human gastrointestinal tract. AB - Zinc homeostasis is primarily maintained via the gastrointestinal system by the processes of absorption of exogenous zinc and gastrointestinal secretion and excretion of endogenous zinc. Although these processes modulate net absorption and the size of the readily exchangeable zinc pools, there are limits to the effectiveness of the homeostatic mechanisms of these and other systems. As a result of the interplay of the subcellular regulation of these mechanisms and host, dietary and environmental factors, zinc deficiency is not uncommon, especially on a global basis. This overview briefly reviews current understanding about the subcellular mechanisms of zinc absorption and transport. Factors recognized to affect zinc absorption at the whole body level are reviewed and include the amount and form of zinc consumed; dietary promoters, such as animal protein and low-molecular-weight organic compounds; dietary inhibitors, such as phytate and possibly iron and calcium when consumed as supplements; and physiologic states, such as pregnancy, lactation and early infancy, all of which increase the demand for absorbed zinc. The control of endogenously secreted zinc is less well understood. Available data suggest that the quantity of secreted zinc with each meal may be considerable and that efficient reabsorption is critical to the maintenance of normal zinc balance. Factors that have been proposed to interfere with the normal reabsorption of endogenous zinc include phytate and unabsorbed fat. Understanding of the dietary, physiologic, pathologic and environmental factors that may adversely affect these processes, and therefore zinc homeostasis, will be critical to preventing and treating zinc deficiency in human populations. PMID- 10801947 TI - Dietary factors influencing zinc absorption. AB - Marginal zinc deficiency and suboptimal zinc status have been recognized in many groups of the population in both less developed and industrialized countries. Although the cause in some cases may be inadequate dietary intake of zinc, inhibitors of zinc absorption are most likely the most common causative factor. Phytate, which is present in staple foods like cereals, corn and rice, has a strong negative effect on zinc absorption from composite meals. Inositol hexaphosphates and pentaphosphates are the phytate forms that exert these negative effects, whereas the lower phosphates have no or little effect on zinc absorption. The removal or reduction of phytate by enzyme (phytase) treatment, precipitation methods, germination, fermentation or plant breeding/genetic engineering markedly improves zinc absorption. Iron can have a negative effect on zinc absorption, if given together in a supplement, whereas no effect is observed when the same amounts are present in a meal as fortificants. Cadmium, which is increasing in the environment, also inhibits zinc absorption. The amount of protein in a meal has a positive effect on zinc absorption, but individual proteins may act differently; e.g., casein has a modest inhibitory effect of zinc absorption compared with other protein sources. Amino acids, such as histidine and methionine, and other low-molecular-weight ions, such as EDTA and organic acids (e.g., citrate), are known to have a positive effect on zinc absorption and have been used for zinc supplements. Knowledge about dietary factors that inhibit zinc absorption and about ways to overcome or remove these factors is essential when designing strategies to improve the zinc nutrition of vulnerable groups. PMID- 10801948 TI - Integrative aspects of zinc transporters. AB - Cells maintain zinc concentrations with relatively narrow limits. Nevertheless, physiologically relevant changes in free Zn(II) pools or changes in Zn bound to specific ligands or within vesicles may occur without a major change in total cellular zinc concentrations. The task of maintaining such levels rests in part with zinc transporter proteins. The genes for some putative zinc transporters have recently been cloned. As of this time, most have not been directly shown to transport zinc in functional studies, albeit evidence is strong that they have such a function. Zinc transporter (ZnT)-1 was identified as a rescue agent for cells maintained in very high extracellular zinc conditions; therefore, ZnT-1 has been suggested to function as an exporter. ZnT-1 is expressed in a variety of tissues, including intestine, kidney and liver. Intestinal expression is regional, being much greater in duodenum and jejunum and in villus versus crypt cells. Immunolocalization places ZnT-1 at the basolateral membrane of intestinal enterocytes and epithelial cells of the distal renal tubules. Regulation of ZnT-1 mRNA and ZnT-1 protein does not change markedly with changes in dietary zinc level except when a large single oral zinc supplement is provided. ZnT-1 is induced by transient ischemia of the forebrain. ZnT-2 and ZnT-3 may function in tissue-specific vesicular zinc transport. ZnT-4 is believed to be abundant in mammary gland and may be associated with zinc secretion into milk. A mutation of the ZnT-4 gene may account for the lethal milk (lm) syndrome. The putative zinc transporters identified thus far appear to have characteristics commensurate with functions in integrative zinc acquisition and homeostasis. PMID- 10801949 TI - Zinc deficiency, malnutrition and the gastrointestinal tract. AB - Recent clinical and experimental findings have reinforced the link among zinc deficiency, malnutrition and diarrheal disease. Because there is a strong association between protein and zinc content in virtually all types of foods, insufficient protein intake may often be the cause of zinc deficiency. Compensatory mechanisms operating in monogastric species during malnutrition are less effective for the absorption of transition divalent elements such as zinc, which remain bound to ligands of dietary or endogenous origin. Both protein and zinc deficiencies are strong negative determinants for normal cellular immunity. In zinc deficiency, the organism is more susceptible to toxin-producing bacteria or enteroviral pathogens that activate guanylate and adenylate cyclases, stimulating chloride secretion, producing diarrhea and diminishing absorption of nutrients, thus exacerbating an already compromised mineral status. In addition, zinc deficiency may impair the absorption of water and electrolytes, delaying the termination of normally self-limiting gastrointestinal disease episodes. The gastrointestinal tract may be one of the first target areas where zinc insufficiency may be manifested. A prolonged low zinc intake deprives the organism of the local potential beneficial effects of zinc, including interactions with oxidative free radicals and nitric oxide metabolism. Nitric oxide is a second messenger that plays an important part in the triggering of diarrheal disease. The possible interrelationship among infection, inflammation, free radical damage and its quenching by potential scavengers, such as zinc, in the intestinal lumen or within the enterocyte should be more extensively studied. PMID- 10801950 TI - Regulation of intestinal gene expression by dietary zinc: induction of uroguanylin mRNA by zinc deficiency. AB - The regulation of gene expression by nutrients plays an important role in the overall manifestations of nutritional deficiencies. Insufficient intakes of dietary micronutrients, such as zinc, produce profound effects in multiple organs and tissues. One of the major challenges, however, is to identify genes affected by changes in nutritional status. Differential display of mRNA has proved to be a valuable technique in meeting this challenge. In our ongoing search for genes responsive to dietary zinc, we compared small intestinal mRNA from rats that were fed zinc-deficient or -adequate diets using differential display to generate 3' anchored expressed sequence tags (EST). EST for intestinal mRNAs with altered expression due to zinc deficiency include two peptide hormones, intestinal fatty acid binding protein, intestinal alkaline phosphatase II, a proteasomal ATPase, cis-Golgi p28 and two subunits of the ubiquinone oxidoreductase. The EST for one of the hormones yielded the sequence for the 3' end of an mRNA encoding preprouroguanylin and was used to clone the remaining portion of the rat cDNA via 5' rapid amplification of cDNA ends. Northern blot analysis of RNA from rat intestine demonstrated that preprouroguanylin mRNA was 2.5-fold more abundant during zinc deficiency. Uroguanylin, a natriuretic peptide hormone, is an endogenous ligand for the same guanylate cyclase C that the Escherichia coli heat stable enterotoxin (STa) binds when it causes secretory diarrhea by activating the cystic fibrosis transmembrane conductance regulator, thus altering fluid balance in the intestine. This suggests a mechanism whereby zinc deficiency could induce uroguanylin levels in the intestine and cause or potentiate diarrhea. PMID- 10801951 TI - The dynamic link between the integrity of the immune system and zinc status. AB - The results of more than three decades of work indicate that zinc deficiency rapidly diminishes antibody- and cell-mediated responses in both humans and animals. The moderate deficiencies in zinc noted in sickle cell anemia, renal disease, chronic gastrointestinal disorders and acrodermatitis enteropathica; subjects with human immunodeficiency virus; children with diarrhea; and elderly persons can greatly alter host defense systems, leading to increases in opportunistic infections and mortality rates. Conversely, short periods of zinc supplementation substantially improve immune defense in individuals with these diseases. Mouse models demonstrate that 30 d of suboptimal intake of zinc can lead to 30-80% losses in defense capacity. Collectively, the data clearly demonstrate that immune integrity is tightly linked to zinc status. Lymphopenia and thymic atrophy, which were the early hallmarks of zinc deficiency, are now known to be due to high losses of precursor T and B cells in the bone marrow. This ultimately leads to lymphopenia or a failure to replenish the lymphocytic system. Glucocorticoid-mediated apoptosis induced by zinc deficiency causes down regulation of lymphopoiesis. Indeed, zinc itself can modulate death processes in precursor lymphocytes. Finally, there is substantial evidence that zinc supplementation may well reduce the impact of many of the aforementioned diseases by preventing the dismantling of the immune system. The latter represents an important area for research. PMID- 10801952 TI - Zinc-altered immune function and cytokine production. AB - Although the intriguing role of zinc as an essential trace element for immune function is well established, particular progress in determining the molecular principles of action of this ion was made recently. Leukocyte responsiveness is delicately regulated by zinc concentration. Zinc deficiency as well as supraphysiologic levels impair immune function. Furthermore, the activities of many immunostimulants frequently used in immunologic studies are influenced by zinc concentration. Therefore, our knowledge from in vitro studies is widely dependent on the zinc concentration, and when not in physiologic range, immunologic responses are artificially low. Decreased production of TH1 cytokines and interferon-alpha by leukocytes in the healthy elderly person is correlated with low zinc serum level. The defect in interferon-alpha production is reconstituted by the addition of physiologic amounts of zinc in vitro. Interestingly, zinc induces cytokine production by isolated leukocytes. Zinc induces monocytes to produce interleukin-1, interleukin-6 and tumor necrosis factor-alpha in peripheral blood mononuclear cells and separated monocytes. This effect is higher in serum-free medium. However, only in the presence of serum does zinc also induce T cells to produce lymphokines. This effect on T cells is mediated by cytokines produced by monocytes. Stimulation also requires cell-to cell contact of monocytes and T cells. Information is presented to illustrate the concepts that the zinc concentration must be taken into account whenever in vitro studies are made or complex alterations of immune functions are observed in vivo. PMID- 10801953 TI - Zinc deficiency impairs immune responses against parasitic nematode infections at intestinal and systemic sites. AB - Research on the complex interactions among host nutritional status, parasitic infection and immune responsiveness has focused on the detrimental consequences of parasitic infections on host nutritional status and on mechanisms by which malnutrition impairs immunocompetence. Curiously, relatively few studies have examined the effects of malnutrition on the immune response in the parasite infected host, and even fewer have considered the events occurring at the intestinal level, where absorption of nutrients occurs, intestinal parasites reside, and the gastrointestinal-associated lymphoid tissues play a role in directing both the local and the more systemic immune responses. Our work using a zinc-deficient nematode-infected mouse model reveals that parasites are better able to survive in the zinc-deficient hosts than in well-nourished hosts; that the production of interleukin-4 in the spleen of zinc-deficient mice is depressed, leading to depressed levels of IgE, IgG(1) and eosinophils; and that the function of T cells and antigen-presenting cells is impaired by zinc deficiency as well as by energy restriction. Given the paramount role of the gastrointestinal-associated lymphoid tissues in inducing and regulating immune responses to intestinal parasites and in orchestrating responses in the spleen and peripheral circulation, we conclude that zinc deficiency (in association with energy restriction) exerts profound effects on the gut mucosal immune system, leading to changes in systemically disseminated immune responses and, importantly, to prolonged parasite survival. PMID- 10801954 TI - Zinc status in human immunodeficiency virus infection. AB - There is substantial evidence to support an important role for zinc in immune processes. Adequate zinc status is essential for T-cell division, maturation and differentiation; lymphocyte response to mitogens; programmed cell death of lymphoid and myeloid origins; gene transcription; and biomembrane function. Lymphocytes are one of the types of cells activated by zinc. Zinc is the structural component of a wide variety of proteins, neuropeptides, hormone receptors and polynucleotides. Among the best known zinc-dependent hormones/enzymes are Cu, Zn superoxide dismutase, an enzyme component of the antioxidant defense system, and thymulin, which is essential for the formation of T-lymphocytes. In animals and humans, zinc deficiency results in rapid and marked atrophy of the thymus, impaired cell-mediated cutaneous sensitivity and lymphopenia. Primary and secondary antibody responses are reduced in zinc deficiency, particularly for those antigens that require T-cell help, such as those in heterologous red blood cells. In addition, antibody response and the generation of splenic cytotoxic T cells after immunization are reduced. Zinc also inhibits the production of tumor necrosis factor, which is implicated in the pathophysiology of cachexia and wasting in acquired immune deficiency syndrome. PMID- 10801955 TI - Therapeutic application of zinc in human immunodeficiency virus against opportunistic infections. AB - The relevance of zinc in resistance to infections by virus, fungi and bacteria is recognized because of its pivotal role in the efficiency of the entire immune system, in particular in conferring biological activity to a thymic hormone called thymulin, which has differentiation properties on T-cell lines. In infection with human immunodeficiency virus (HIV), the zinc-bound form of thymulin (active thymulin, ZnFTS) is strongly reduced in stage IV of the disease (Centers for Disease Control and Prevention classification) with concomitant decrements in CD4(+) cell count and zincemia values. The zinc-unbound form of thymulin (inactive thymulin, FTS) is, in contrast, very high. The in vitro addition of zinc to plasma samples induces a recovery of the thymulin active form, suggesting low zinc bioavailability as the cause of impaired thymic functions with consequent CD4(+) depletion. An analysis of risk factors for the incidence of recidivism opportunistic infections shows CD4(+) depletion and zinc deficiency to have significant scores. Supplementation with zinc for 1 mo (45 mg Zn(2+)/d) associated with zidovudine (AZT) therapy in stage IV induces recovery of active zinc-bound thymulin, of zincemia, of CD4(+) cells with concomitant reduction (50%) of recidivism opportunistic infections compared with the AZT treated group. Complete disappearance of recidivism by Candida aesophagea or Pneumocystis carinii is observed after supplementation with zinc. The relative risk factors (CD4(+) depletion and zinc-deficiency) have lower scores in the HIV positive zinc-treated group, confirming, as such, the relevance of zinc in opportunistic infections that involve extracellular matrix. Such an assumption is indirectly confirmed with new HAART, where no opportunistic infections occur. Indeed, HIV RNA is inversely correlated with both CD4(+) and zincemia values (r = -0.73, P<0.01) in HAART-treated subjects. Lower scores for the same relative factors for the appearance of opportunistic infections are present in HAART treated subjects compared with those treated with AZT. These findings, on the one hand, show the poor efficacy of AZT therapy compared with HAART therapy for the progression of HIV, but on the other hand, they suggest that the lack of occurrence of opportunistic infections by HAART may also result from major zinc bioavailability. This further supports the key role played by zinc against opportunistic infections in HIV with a possible independent effect by either HIV or the pathogens involved. PMID- 10801956 TI - Role of zinc in plasma membrane function. AB - The concentration of plasma zinc is the generally accepted index of zinc status. Although low plasma zinc is an essential criterion of deficiency, alone it is inadequate. To supplement this index, we sought to determine the first limiting biochemical defect in animals fed zinc-deficient diets and concluded that the limiting function is associated with a posttranslational change in plasma membrane proteins. Among the signs of zinc deficiency in rats is a bleeding tendency associated with failure of platelet aggregation, a phenomenon that correlates with impaired uptake of Ca(2+) when stimulated. Zinc-deficient guinea pigs exhibit signs of peripheral neuropathy, and their brain synaptic vesicles exhibit impaired Ca(2+) uptake when they are stimulated with glutamate. Red cells from zinc-deficient rats show increased osmotic fragility associated with decreased plasma membrane sulfhydryl concentration. Both phenomena are readily reversed (2 d) by dietary zinc repletion. Volume recovery is dependent on Ca dependent K channels and the sulfhydryl redox state. Both the impaired aggregation and calcium uptake of zinc-deficient platelets are corrected by in vitro incubation of blood with glutathione. Considering the fact that plasma membranes from several cell types show impaired function that is associated with a decreased rate of calcium uptake, it is postulated that a defect in calcium channels is the first limiting biochemical defect in zinc deficiency. The calcium uptake defect and consequent impaired second-messenger function likely results from an abnormal sulfhydryl redox state in the membrane channel protein. PMID- 10801957 TI - Function and mechanism of zinc metalloenzymes. AB - Zinc is required for the activity of > 300 enzymes, covering all six classes of enzymes. Zinc binding sites in proteins are often distorted tetrahedral or trigonal bipyramidal geometry, made up of the sulfur of cysteine, the nitrogen of histidine or the oxygen of aspartate and glutamate, or a combination. Zinc in proteins can either participate directly in chemical catalysis or be important for maintaining protein structure and stability. In all catalytic sites, the zinc ion functions as a Lewis acid. Researchers in our laboratory are dissecting the determinants of molecular recognition and catalysis in the zinc-binding site of carbonic anhydrase. These studies demonstrate that the chemical nature of the direct ligands and the structure of the surrounding hydrogen bond network are crucial for both the activity of carbonic anhydrase and the metal ion affinity of the zinc-binding site. An understanding of naturally occurring zinc-binding sites will aid in creating de novo zinc-binding proteins and in designing new metal sites in existing proteins for novel purposes such as to serve as metal ion biosensors. PMID- 10801958 TI - The antioxidant properties of zinc. AB - The ability of zinc to retard oxidative processes has been recognized for many years. In general, the mechanism of antioxidation can be divided into acute and chronic effects. Chronic effects involve exposure of an organism to zinc on a long-term basis, resulting in induction of some other substance that is the ultimate antioxidant, such as the metallothioneins. Chronic zinc deprivation generally results in increased sensitivity to some oxidative stress. The acute effects involve two mechanisms: protection of protein sulfhydryls or reduction of (*)OH formation from H(2)O(2) through the antagonism of redox-active transition metals, such as iron and copper. Protection of protein sulfhydryl groups is thought to involve reduction of sulfhydryl reactivity through one of three mechanisms: (1) direct binding of zinc to the sulfhydryl, (2) steric hindrance as a result of binding to some other protein site in close proximity to the sulfhydryl group or (3) a conformational change from binding to some other site on the protein. Antagonism of redox-active, transition metal-catalyzed, site specific reactions has led to the theory that zinc may be capable of reducing cellular injury that might have a component of site-specific oxidative damage, such as postischemic tissue damage. Zinc is capable of reducing postischemic injury to a variety of tissues and organs through a mechanism that might involve the antagonism of copper reactivity. Although the evidence for the antioxidant properties of zinc is compelling, the mechanisms are still unclear. Future research that probes these mechanisms could potentially develop new antioxidant functions and uses for zinc. PMID- 10801959 TI - The function of zinc metallothionein: a link between cellular zinc and redox state. AB - A chemical and biochemical mechanism of action of the metallothionein (MT)/thionein (T) couple has been proposed. The mechanism emphasizes the importance of zinc/sulfur cluster bonding in MT and the significance of the two cluster networks as redox units that confer mobility on otherwise tightly bound and redox-inert zinc in MT. In this article, it is further explored how this redox mechanism controls the metabolically active cellular zinc pool. The low redox potential of the sulfur donor atoms in the clusters readily allows oxidation by mild cellular oxidants with concomitant release of zinc. Such a release by oxidants and the preservation of zinc binding by antioxidants place MT under the control of the cellular redox state and, consequently, energy metabolism. The binding of effectors, e.g., ATP, elicits conformational changes and alters zinc binding in MT. The glutathione/glutathione disulfide redox couple as well as selenium compounds effect zinc delivery from MT to the apoforms of zinc enzymes. This novel action of selenium on zinc/sulfur coordination sites has significant implications for the interaction between these essential elements. Tight binding and kinetic lability, modulation of MT by cellular ligands and the redox state, control of MT gene expression by zinc and many other inducers all support a critical function of the MT/T system in cellular homeostasis and distribution of zinc. PMID- 10801960 TI - Cellular zinc fluxes and the regulation of apoptosis/gene-directed cell death. AB - The maintenance of discrete subcellular pools of zinc (Zn) is critical for the functional and structural integrity of cells. Among the important biological processes influenced by Zn is apoptosis, a process that is important in cellular homeostasis (an important cellular homeostatic process). It has also been identified as a major mechanism contributing to cell death in response to toxins and in disease, offering hope that novel therapies that target apoptotic pathways may be developed. Because Zn levels in the body can be increased in a relatively nontoxic manner, it may be possible to prevent or ameliorate degenerative disorders that are associated with high rates of apoptotic cell death. This review begins with brief introductions that address, first, the cellular biology of Zn, especially the critical labile Zn pools, and, second, the phenomenon of apoptosis. We then review the evidence relating Zn to apoptosis and address three major hypotheses: (1) that a specific pool or pools of intracellular labile Zn regulates apoptosis; (2) that systemic changes in Zn levels in the body, due to dietary factors, altered physiological states or disease, can influence cell susceptibility to apoptosis, and (3) that this altered susceptibility to apoptosis contributes to pathophysiological changes in the body. Other key issues are the identity of the molecular targets of Zn in the apoptotic cascade, the types of cells and tissues most susceptible to Zn-regulated apoptosis, the role of Zn as a coordinate regulator of mitosis and apoptosis and the apparent release of tightly bound intracellular pools of Zn during the later stages of apoptosis. This review concludes with a section highlighting areas of priority for future studies. PMID- 10801961 TI - Metallothionein, nitric oxide and zinc homeostasis in vascular endothelial cells. AB - Recent in vitro studies suggest that the oxidoreductive capacity of metal thiolate clusters in metallothionein (MT) contributes to intracellular zinc homeostasis. We used fluorescence-based techniques to address this hypothesis in intact endothelial cells, focusing on the contributory role of the important redox signaling molecule, nitric oxide. Microspectrofluorometry with Zinquin revealed that the exposure of cultured sheep pulmonary artery endothelial cells to S-nitrosocysteine resulted in the release of N, N,N',N'-tetrakis(2. pyridylmethyl)ethylendiamine (TPEN) chelatable zinc. Cultured sheep pulmonary artery endothelial cells were transfected with a plasmid expression vector suitable for fluorescence resonance energy transfer containing the cDNA of MT sandwiched between two mutant green fluorescent proteins. The exposure of cultured sheep pulmonary artery endothelial cells transfected with this chimera to nitric oxide donors or to agents that increased cytoplasmic Ca(2+) via endogenously generated nitric oxide decreased the efficiency of fluorescence resonance energy transfer in a manner consistent with the release of metal (Zn) from MT. A physiological role for this interaction in intact tissue was supported by the lack of myogenic reflex in resistance arteries of MT knockout mice unless endogenous nitric oxide synthesis was blocked. These data suggest an important role for metal thiolate clusters of MT in nitric oxide signaling in the vascular wall. PMID- 10801962 TI - Importance of zinc in the central nervous system: the zinc-containing neuron. AB - Zinc is essential to the structure and function of myriad proteins, including regulatory, structural and enzymatic. It is estimated that up to 1% of the human genome codes for zinc finger proteins. In the central nervous system, zinc has an additional role as a neurosecretory product or cofactor. In this role, zinc is highly concentrated in the synaptic vesicles of a specific contingent of neurons, called "zinc-containing" neurons. Zinc-containing neurons are a subset of glutamatergic neurons. The zinc in the vesicles probably exceeds 1 mmol/L in concentration and is only weakly coordinated with any endogenous ligand. Zinc containing neurons are found almost exclusively in the forebrain, where in mammals they have evolved into a complex and elaborate associational network that interconnects most of the cerebral cortices and limbic structures. Indeed, one of the intriguing aspects of these neurons is that they compose somewhat of a chemospecific "private line" of the mammalian cerebral cortex. The present review outlines (1) the methods used to discover, define and describe zinc-containing neurons; (2) the neuroarchitecture and synaptology of zinc-containing neural circuits; (3) the physiology of regulated vesicular zinc release; (4) the "life cycle" and molecular biology of vesicular zinc; (5) the importance of synaptically released zinc in the normal and pathological processes of the cerebral cortex; and (6) the role of specific and nonspecific stressors in the release of zinc. PMID- 10801963 TI - Zinc transport in the brain: routes of zinc influx and efflux in neurons. AB - Studies of the routes of entry and exit for zinc in different tissues and cell types have shown that zinc can use several pathways of exit or entry. In neurons, known pathways include (1) presynaptic release along with glutamate when synaptic vesicles empty their contents into the synaptic cleft, (2) voltage-gated L-type Ca(2+) channels and glutamate-gated channels that provide an entry route when cells are depolarized and that mediate extracellular zinc toxicity and (3) a plasma membrane transporter potentially present in all neurons important for cellular zinc homeostasis. The least understood of these pathways, in terms of mechanism, is the transporter pathway. The kinetics of zinc uptake in cultured neurons under resting conditions are consistent with and suggest the existence of a saturable transporter in the plasma membrane. The proteins responsible for plasma membrane zinc transport have not yet been definitely identified. Likely candidates include two proteins identified by molecular cloning termed zinc transporter 1 and divalent cation transporter DCT1. Both proteins have been shown to be expressed in the brain, but only DCT1 is clearly demonstrated to be a transport protein, whereas zinc transporter 1 may only modulate zinc transport in association with as-yet-unidentified proteins. Understanding the mechanism and neuromodulation of plasma membrane zinc transport will be an important first step toward a complete understanding of neuronal zinc homeostasis. PMID- 10801964 TI - Alzheimer's disease, beta-amyloid protein and zinc. AB - Alzheimer's disease (AD) is characterized by amyloid deposits within the neocortical parenchyma and the cerebrovasculature. The main component of these predominantly extracellular collections, Abeta, which is normally a soluble component of all biological fluids, is cleaved out of a ubiquitously expressed parent protein, the amyloid protein precursor (APP), one of the type 1 integral membrane glycoproteins. Considerable evidence has indicated that there is zinc dyshomeostasis and abnormal cellular zinc mobilization in AD. We have characterized both APP and Abeta as copper/zinc metalloproteins. Zinc, copper and iron have recently been reported to be concentrated to 0.5 to 1 mmol/L in amyloid plaque. In vitro, rapid Abeta aggregation is mediated by Zn(II), promoted by the alpha-helical structure of Abeta, and is reversible with chelation. In addition, Abeta produces hydrogen peroxide in a Cu(II)/Fe(III)-dependent manner, and the hydrogen peroxide formation is quenched by Zn(II). Moreover, zinc preserves the nontoxic properties of Abeta. Although the zinc-binding proteins apolipoprotein E epsilon4 allele and alpha(2)-macroglobulin have been characterized as two genetic risk factors for AD, zinc exposure as a risk factor for AD has not been rigorously studied. Based on our findings, we envisage that zinc may serve twin roles by both initiating amyloid deposition and then being involved in mechanisms attempting to quench oxidative stress and neurotoxicity derived from the amyloid mass. Hence, it remains debatable whether zinc supplementation is beneficial or deleterious for AD until additional studies clarify the issue. PMID- 10801965 TI - Neurobiology of zinc-influenced eating behavior. AB - Zinc is an essential nutrient that is required in humans and animals for many physiological functions, including immune and antioxidant function, growth and reproduction. Many aspects of zinc deficiency-induced anorexia have been well studied in experimental animals, most notably the laboratory rat. There is evidence that suggests zinc deficiency may be intimately involved with anorexia in humans: if not as an initiating cause, then as an accelerating or exacerbating factor that may deepen the pathology of the anorexia. The present review describes recent research investigating the relationship between zinc deficiency and the regulation of food intake, along with advances in the understanding of the food intake and body weight regulation systems. For more comprehensive reviews of zinc nutrition and zinc deficiency, readers are referred to the other reviews in this volume and the review text of Mills (1989). An excellent review focused solely on zinc status and food intake has been presented by O'Dell and Reeves (1989). PMID- 10801966 TI - The role of zinc in growth and cell proliferation. AB - The inhibition of growth is a cardinal symptom of zinc deficiency. In animals fed a zinc-inadequate diet, both food intake and growth are reduced within 4-5 d. Despite the concomitant reduction in food intake and growth, reduced energy intake is not the limiting factor in growth, because force-feeding a zinc inadequate diet to animals fails to maintain growth. Hence, food intake and growth appear to be regulated by zinc through independent, although well coordinated, mechanisms. Despite the long-term study of zinc metabolism, the first limiting role of zinc in cell proliferation remains undefined. Zinc participates in the regulation of cell proliferation in several ways; it is essential to enzyme systems that influence cell division and proliferation. Removing zinc from the extracellular milieu results in decreased activity of deoxythymidine kinase and reduced levels of adenosine(5')tetraphosphate(5') adenosine. Hence, zinc may directly regulate DNA synthesis through these systems. Zinc also influences hormonal regulation of cell division. Specifically, the pituitary growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis is responsive to zinc status. Both increased and decreased circulating concentrations of GH have been observed in zinc deficiency, although circulating IGF-I concentrations are consistently decreased. However, growth failure is not reversed by maintaining either GH or IGF-I levels through exogenous administration, which suggests the defect occurs in hormone signaling. Zinc appears to be essential for IGF-I induction of cell proliferation; the site of regulation is postreceptor binding. Overall, the evidence suggests that reduced zinc availability affects membrane signaling systems and intracellular second messengers that coordinate cell proliferation in response to IGF-I. PMID- 10801967 TI - Zinc in relation to diabetes and oxidative disease. AB - Theoretically, zinc can exert a number of indirect antioxidant functions. Researchers at our laboratory have found evidence to support this concept by studying mild zinc deficiency in rats. This state produces low resistance to chemically induced liver oxidant injury, and it produces high vulnerability of lipoproteins to oxidation. We are building on this work in rats to test a hypothesis in humans that increased zinc intake will protect against oxidant stress in persons with tendencies for both moderate zinc deficiency and high oxidant stress. This hypothesis has been tested in postmenopausal, type 2 diabetic women. A 3-wk supplementation with zinc (30 mg/d as glycine-chelate) raised initially low plasma zinc values to above normal values and increased plasma activities of 5'-nucleotidase. However, the latter values were still well below normal. Lipoprotein oxidation tendencies, a measure of oxidant stress, were not altered by the zinc treatment. A new project has been initiated to determine whether both a higher dose and longer duration of zinc treatment will normalize 5'-nucleotidase activities and affect the indices of oxidant stress. The latter will be considered in terms of both zinc supplementation and supplementation of zinc plus vitamin C, another problem nutrient for diabetic persons. PMID- 10801968 TI - Zinc and the common cold: a meta-analysis revisited. AB - The common cold has been estimated to cost the United States > $3.5 billion per year. Despite several randomized clinical trials, the effect of treating colds with zinc gluconate remains uncertain due to conflicting results. We conducted a meta-analysis of published randomized clinical trials on the use of zinc gluconate lozenges in colds using the random effects model of DerSimonians and Laird. Ten clinical trials of cold treatment with zinc gluconate were identified. After excluding two studies that used nasal inoculum of rhinovirus, eight trials were combined and analyzed. The summary odds ratio for the presence of "any cold symptoms" at 7 d was 0.52 (95% confidence interval, 0.25-1.2). We conclude that despite numerous randomized trials, the evidence for effectiveness of zinc lozenges in reducing the duration of common colds is still lacking. PMID- 10801969 TI - The Age-Related Eye Disease Study: a clinical trial of zinc and antioxidants--Age Related Eye Disease Study Report No. 2. AB - The Age-Related Eye Disease Study includes a clinical trial of high dose antioxidant and zinc supplements for age-related macular degeneration and a clinical trial of high dose antioxidants for cataract. The clinical trials were initiated largely because of the widespread public use in the United States of commercially available pharmacological doses of vitamins and minerals to treat these two eye conditions and the absence of definitive studies on the safety and efficacy of their use. This report focuses on the clinical trial of zinc and the possibility that this elderly study cohort, who were randomly assigned in a factorial design to receive zinc, antioxidants and placebo, could be assessed for zinc effects other than those currently addressed by the trial. PMID- 10801970 TI - Participants in the workshop "Zinc and health: current status and future Directions" held november 4-5, 1998 at the national institutes of health in bethesda, MD PMID- 10801971 TI - Dynamic interplay between antagonistic pathways controlling the sigma 32 level in Escherichia coli. AB - The heat-shock response in Escherichia coli depends primarily on the transient increase in the cellular level of heat-shock sigma factor final sigma(32) encoded by the rpoH gene, which results from both enhanced synthesis and transient stabilization of normally unstable final sigma(32). Heat-induced synthesis of final sigma(32) was previously shown to occur at the translation level by melting the mRNA secondary structure formed within the 5' coding sequence of rpoH including the translation initiation region. The subsequent decrease in the final sigma(32) level during the adaptation phase has been thought to involve both shutoff of synthesis (translation) and destabilization of final sigma(32) mediated by the DnaK-DnaJ chaperones, although direct evidence for translational repression was lacking. We now show that the heat-induced synthesis of final sigma(32) does not shut off at the translation level by using a reporter system involving translational coupling. Furthermore, the apparent shutoff was not observed when the synthesis rate was determined by a very short pulse labeling (15 s). Examination of final sigma(32) stability at 10 min after shift from 30 to 42 degrees C revealed more extreme instability (t(1/2)=20 s) than had previously been thought. Thus, the dynamic change in final sigma(32) stability during the heat-shock response largely accounts for the apparent shutoff of final sigma(32) synthesis observed with a longer pulse. These results suggest a mechanism for maintaining the intricate balance between the antagonistic pathways: the rpoH translation as determined primarily by ambient temperature and the turnover of final sigma(32) as modulated by the chaperone (and presumably protease)-mediated autogenous control. PMID- 10801972 TI - Crystallographic snapshots along a protein-induced DNA-bending pathway. AB - Two new high-resolution cocrystal structures of EcoRV endonuclease bound to DNA show that a large variation in DNA-bending angles is sampled in the ground state binary complex. Together with previous structures, these data reveal a contiguous series of protein conformational states delineating a specific trajectory for the induced-fit pathway. Rotation of the DNA-binding domains, together with movements of two symmetry-related helices binding in the minor groove, causes base unstacking at a key base-pair step and propagates structural changes that assemble the active sites. These structures suggest a complex mechanism for DNA bending that depends on forces generated by interacting protein segments, and on selective neutralization of phosphate charges along the inner face of the bent double helix. PMID- 10801973 TI - A phage integrase directs efficient site-specific integration in human cells. AB - The integrase from the Streptomyces phage phiC31 carries out efficient recombination between the attP site in the phage genome and the attB site in the host bacterial chromosome. In this paper, we show that the enzyme also functions in human cells. A plasmid assay system was constructed that measured intramolecular integration of attP into attB. This assay was used to demonstrate that in the presence of the phiC31 integrase, precise unidirectional integration occurs with an efficiency of 100% in Escherichia coli and >50% in human cells. This assay system was also used to define the minimal sizes of attB and attP at 34 bp and 39 bp, respectively. Furthermore, precise and efficient intermolecular integration of an incoming plasmid bearing attP into an established Epstein-Barr virus plasmid bearing attB was documented in human cells. This work is a demonstration of efficient, site-specific, unidirectional integration in mammalian cells. These observations form the basis for site-specific integration strategies potentially useful in a broad range of genetic engineering applications. PMID- 10801974 TI - Structure and mechanism of mammalian thioredoxin reductase: the active site is a redox-active selenolthiol/selenenylsulfide formed from the conserved cysteine selenocysteine sequence. AB - Mammalian thioredoxin reductases (TrxR) are homodimers, homologous to glutathione reductase (GR), with an essential selenocysteine (SeCys) residue in an extension containing the conserved C-terminal sequence -Gly-Cys-SeCys-Gly. In the oxidized enzyme, we demonstrated two nonflavin redox centers by chemical modification and peptide sequencing: one was a disulfide within the sequence -Cys(59)-Val-Asn-Val Gly-Cys(64), identical to the active site of GR; the other was a selenenylsulfide formed from Cys(497)-SeCys(498) and confirmed by mass spectrometry. In the NADPH reduced enzyme, these centers were present as a dithiol and a selenolthiol, respectively. Based on the structure of GR, we propose that in TrxR, the C terminal Cys(497)-SeCys(498) residues of one monomer are adjacent to the Cys(59) and Cys(64) residues of the second monomer. The reductive half-reaction of TrxR is similar to that of GR followed by exchange from the nascent Cys(59) and Cys(64) dithiol to the selenenylsulfide of the other subunit to generate the active-site selenolthiol. Characterization of recombinant mutant rat TrxR with SeCys(498) replaced by Cys having a 100-fold lower k(cat) for Trx reduction revealed the C-terminal redox center was present as a dithiol when the Cys(59) Cys(64) was a disulfide, demonstrating that the selenium atom with its larger radius is critical for formation of the unique selenenylsulfide. Spectroscopic redox titrations with dithionite or NADPH were consistent with the structure model. Mechanisms of TrxR in reduction of Trx and hydroperoxides have been postulated and are compatible with known enzyme activities and the effects of inhibitors, like goldthioglucose and 1-chloro-2,4-dinitrobenzene. PMID- 10801975 TI - Jewish and Middle Eastern non-Jewish populations share a common pool of Y chromosome biallelic haplotypes. AB - Haplotypes constructed from Y-chromosome markers were used to trace the paternal origins of the Jewish Diaspora. A set of 18 biallelic polymorphisms was genotyped in 1,371 males from 29 populations, including 7 Jewish (Ashkenazi, Roman, North African, Kurdish, Near Eastern, Yemenite, and Ethiopian) and 16 non-Jewish groups from similar geographic locations. The Jewish populations were characterized by a diverse set of 13 haplotypes that were also present in non-Jewish populations from Africa, Asia, and Europe. A series of analyses was performed to address whether modern Jewish Y-chromosome diversity derives mainly from a common Middle Eastern source population or from admixture with neighboring non-Jewish populations during and after the Diaspora. Despite their long-term residence in different countries and isolation from one another, most Jewish populations were not significantly different from one another at the genetic level. Admixture estimates suggested low levels of European Y-chromosome gene flow into Ashkenazi and Roman Jewish communities. A multidimensional scaling plot placed six of the seven Jewish populations in a relatively tight cluster that was interspersed with Middle Eastern non-Jewish populations, including Palestinians and Syrians. Pairwise differentiation tests further indicated that these Jewish and Middle Eastern non-Jewish populations were not statistically different. The results support the hypothesis that the paternal gene pools of Jewish communities from Europe, North Africa, and the Middle East descended from a common Middle Eastern ancestral population, and suggest that most Jewish communities have remained relatively isolated from neighboring non-Jewish communities during and after the Diaspora. PMID- 10801976 TI - Altered pain responses in mice lacking alpha 1E subunit of the voltage-dependent Ca2+ channel. AB - alpha(1) subunit of the voltage-dependent Ca(2+) channel is essential for channel function and determines the functional specificity of various channel types. alpha(1E) subunit was originally identified as a neuron-specific one, but the physiological function of the Ca(2+) channel containing this subunit (alpha(1E) Ca(2+) channel) was not clear compared with other types of Ca(2+) channels because of the limited availability of specific blockers. To clarify the physiological roles of the alpha(1E) Ca(2+) channel, we have generated alpha(1E) mutant (alpha(1E)-/-) mice by gene targeting. The lacZ gene was inserted in-frame and used as a marker for alpha(1E) subunit expression. alpha(1E)-/- mice showed reduced spontaneous locomotor activities and signs of timidness, but other general behaviors were apparently normal. As involvement of alpha(1E) in pain transmission was suggested by localization analyses with 5-bromo-4-chloro-3 indolyl beta-d-galactopyranoside staining, we conducted several pain-related behavioral tests using the mutant mice. Although alpha(1E)+/- and alpha(1E)-/- mice exhibited normal pain behaviors against acute mechanical, thermal, and chemical stimuli, they both showed reduced responses to somatic inflammatory pain. alpha(1E)+/- mice showed reduced response to visceral inflammatory pain, whereas alpha(1E)-/- mice showed apparently normal response compared with that of wild-type mice. Furthermore, alpha(1E)-/- mice that had been presensitized with a visceral noxious conditioning stimulus showed increased responses to a somatic inflammatory pain, in marked contrast with the wild-type mice in which long lasting effects of descending antinociceptive pathway were predominant. These results suggest that the alpha(1E) Ca(2 +) channel controls pain behaviors by both spinal and supraspinal mechanisms. PMID- 10801977 TI - Reaction coordinates of biomolecular isomerization. AB - Transition path sampling has been applied to the molecular dynamics of the alanine dipeptide in vacuum and in aqueous solution. The analysis shows that more degrees of freedom than the traditional dihedral angles, phi and psi, are necessary to describe the reaction coordinates for isomerization of this molecule. In vacuum, an additional dihedral angle is identified as significant. In solution, solvent variables are shown to play a significant role, and this role appears to be more specific than can be captured by friction models. Implications for larger molecules are discussed. PMID- 10801978 TI - A DNA transfection system for generation of influenza A virus from eight plasmids. AB - We have developed an eight-plasmid DNA transfection system for the rescue of infectious influenza A virus from cloned cDNA. In this plasmid-based expression system, viral cDNA is inserted between the RNA polymerase I (pol I) promoter and terminator sequences. This entire pol I transcription unit is flanked by an RNA polymerase II (pol II) promoter and a polyadenylation site. The orientation of the two transcription units allows the synthesis of negative-sense viral RNA and positive-sense mRNA from one viral cDNA template. This pol I-pol II system starts with the initiation of transcription of the two cellular RNA polymerase enzymes from their own promoters, presumably in different compartments of the nucleus. The interaction of all molecules derived from the cellular and viral transcription and translation machinery results in the generation of infectious influenza A virus. The utility of this system is proved by the recovery of the two influenza A viruses: A/WSN/33 (H1N1) and A/Teal/HK/W312/97 (H6N1). Seventy two hours after the transfection of eight expression plasmids into cocultured 293T and MDCK cells, the virus yield in the supernatant of the transfected cells was between 2 x 10(5) and 2 x 10(7) infectious viruses per milliliter. We also used this eight-plasmid system for the generation of single and quadruple reassortant viruses between A/Teal/HK/W312/97 (H6N1) and A/WSN/33 (H1N1). Because the pol I-pol II system facilitates the design and recovery of both recombinant and reassortant influenza A viruses, it may also be applicable to the recovery of other RNA viruses entirely from cloned cDNA. PMID- 10801979 TI - Specific phenotypic restoration of an attenuated virus by knockout of a host resistance gene. AB - To produce disease, viruses must enter the host, multiply locally in host tissues, spread from the site of entry, and overcome or evade host immune responses. At each stage in this infectious process, specific microbial and host genes determine the ultimate virulence of the virus. Genetic approaches have identified many viral genes that play critical roles in virulence and are presumed to target specific components of the host innate and acquired immune response. However, formal proof that a virulence gene targets a specific protein in a host pathway in vivo has not been obtained. Based on cell culture studies, it has been proposed that the herpes simplex virus type 1 gene ICP34.5 (ICP, infected cell protein) enhances neurovirulence by negating antiviral functions of the IFN-inducible double-stranded RNA-dependent protein kinase R or PKR [Chou, J., Chen, J.J., Gross, M. & Roizman, B. (1995) Proc. Natl. Acad. Sci. USA 92, 10516-10520]. Herein, we show that a virus that has been attenuated by deletion of ICP34.5 exhibits wild-type replication and virulence in a host from which the PKR gene has been deleted. We show that restoration of virulence is specific to ICP34.5 and PKR by using additional host and viral mutants. The use of recombinant viruses to infect animals with null mutations in host defense genes provides a formal genetic test for identifying in vivo mechanisms and targets of microbial virulence genes. PMID- 10801980 TI - Increased metalloproteinase activity, oxidant production, and emphysema in surfactant protein D gene-inactivated mice. AB - Targeted ablation of the surfactant protein D (SP-D) gene caused chronic inflammation, emphysema, and fibrosis in the lungs of SP-D (-/-) mice. Although lung morphology was unperturbed during the first 2 weeks of life, airspace enlargement was observed by 3 weeks and progressed with advancing age. Inflammation consisted of hypertrophic alveolar macrophages and peribronchiolar perivascular monocytic infiltrates. These abnormalities were associated with increased activity of the matrix metalloproteinases, MMP2 and MMP9, and immunostaining for MMP9 and MMP12 in alveolar macrophages. Hydrogen peroxide production by isolated alveolar macrophages also was increased significantly (10 fold). SP-D plays a critical role in the suppression of alveolar macrophage activation, which may contribute to the pathogenesis of chronic inflammation and emphysema. PMID- 10801981 TI - The O-GlcNAc transferase gene resides on the X chromosome and is essential for embryonic stem cell viability and mouse ontogeny. AB - Nuclear and cytoplasmic protein glycosylation is a widespread and reversible posttranslational modification in eukaryotic cells. Intracellular glycosylation by the addition of N-acetylglucosamine (GlcNAc) to serine and threonine is catalyzed by the O-GlcNAc transferase (OGT). This "O-GlcNAcylation" of intracellular proteins can occur on phosphorylation sites, and has been implicated in controlling gene transcription, neurofilament assembly, and the emergence of diabetes and neurologic disease. To study OGT function in vivo, we have used gene-targeting approaches in male embryonic stem cells. We find that OGT mutagenesis requires a strategy that retains an intact OGT gene as accomplished by using Cre-loxP recombination, because a deletion in the OGT gene results in loss of embryonic stem cell viability. A single copy of the OGT gene is present in the male genome and resides on the X chromosome near the centromere in region D in the mouse spanning markers DxMit41 and DxMit95, and in humans at Xq13, a region associated with neurologic disease. OGT RNA expression in mice is comparably high among most cell types, with lower levels in the pancreas. Segregation of OGT alleles in the mouse germ line with ZP3-Cre recombination in oocytes reveals that intact OGT alleles are required for completion of embryogenesis. These studies illustrate the necessity of conditional gene targeting approaches in the mutagenesis and study of essential sex-linked genes, and indicate that OGT participation in intracellular glycosylation is essential for embryonic stem cell viability and for mouse ontogeny. PMID- 10801982 TI - Beyond re-membering: phantom sensations of congenitally absent limbs. AB - Phantom limbs are traditionally conceptualized as the phenomenal persistence of a body part after deafferentation. Previous clinical observations of subjects with phantoms of congenitally absent limbs are not compatible with this view, but, in the absence of experimental work, the neural basis of such "aplasic phantoms" has remained enigmatic. In this paper, we report a series of behavioral, imaging, and neurophysiological experiments with a university-educated woman born without forearms and legs, who experiences vivid phantom sensations of all four limbs. Visuokinesthetic integration of tachistoscopically presented drawings of hands and feet indicated an intact somatic representation of these body parts. Functional magnetic resonance imaging of phantom hand movements showed no activation of primary sensorimotor areas, but of premotor and parietal cortex bilaterally. Movements of the existing upper arms produced activation expanding into the hand territories deprived of afferences and efferences. Transcranial magnetic stimulation of the sensorimotor cortex consistently elicited phantom sensations in the contralateral fingers and hand. In addition, premotor and parietal stimulation evoked similar phantom sensations, albeit in the absence of motor evoked potentials in the stump. These data indicate that body parts that have never been physically developed can be represented in sensory and motor cortical areas. Both genetic and epigenetic factors, such as the habitual observation of other people moving their limbs, may contribute to the conscious experience of aplasic phantoms. PMID- 10801984 TI - PREFACE. PMID- 10801983 TI - Presenilin 1 is linked with gamma-secretase activity in the detergent solubilized state. AB - gamma-Secretase is a membrane-associated protease that cleaves within the transmembrane region of amyloid precursor protein to generate the C termini of the two Abeta peptide isoforms, Abeta40 and Abeta42. Here we report the detergent solubilization and partial characterization of gamma-secretase. The activity of solubilized gamma-secretase was measured with a recombinant substrate, C100Flag, consisting largely of the C-terminal fragment of amyloid precursor protein downstream of the beta-secretase cleavage site. Cleavage of C100Flag by gamma secretase was detected by electrochemiluminescence using antibodies that specifically recognize the Abeta40 or Abeta42 termini. Incubation of C100Flag with HeLa cell membranes or detergent-solubilized HeLa cell membranes generates both the Abeta40 and Abeta42 termini. Recovery of catalytically competent, soluble gamma-secretase critically depends on the choice of detergent; CHAPSO (3 [(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate) but not Triton X-100 is suitable. Solubilized gamma-secretase activity is inhibited by pepstatin and more potently by a novel aspartyl protease transition-state analog inhibitor that blocks formation of Abeta40 and Abeta42 in mammalian cells. Upon gel exclusion chromatography, solubilized gamma-secretase activity coelutes with presenilin 1 (PS1) at an apparent relative molecular weight of approximately 2.0 x 10(6). Anti-PS1 antibody immunoprecipitates gamma-secretase activity from the solubilized gamma-secretase preparation. These data suggest that gamma-secretase activity is catalyzed by a PS1-containing macromolecular complex. PMID- 10801985 TI - Facilitating Development of Multiple-Species Conservation Reserves and Habitat Conservation Plans: A Synthesis of Recommendations. AB - The purpose of this paper is to outline briefly the process that workshop participants undertook to first identify and then resolve major issues hindering the development and implementation of MS-HCPs. In addition, a brief outline of PMID- 10801986 TI - Habitat Conservation Plans Under the U.S. Endangered Species Act: The Legal Perspective. AB - Habitat Conservation Plans under the federal Endangered Species Act have become an increasingly popular tool for resolving conflicts between land development and species conservation. Their primary purpose, however, is legal and regulatory rather than biological. They are what landowners must prepare in order to obtain a permit to "take" animals listed by the U.S. Fish and Wildlife Service as threatened or endangered. Unfortunately, many professionals involved in the HCP process aren't sufficiently cognizant of the legal and regulatory functions and the purposes and limitations of HCPs. I provide an overview of the regulatory structure of the ESA, the role HCPs play in that structure, and the specific legal requirements associated with HCPs. I then discuss the practice of crafting an HCP and the most common issues that arise in the process. Finally, I assess several very fundamental current problems with the HCP program, problems that threaten to undermine the HCP program to such a degree as to end its utility to landowners and thereby end the tremendous conservation opportunities the HCP program represents. PMID- 10801987 TI - Multiple Species Habitat Conservation Planning: Goals and Strategies of Local Governments. AB - Drawing from experiences gained from the development and implementation of four approved habitat conservation plans (HCPs), I describe the goals and strategies used by the nine local government members of the Riverside County Habitat Conservation Agency (RCHCA) to reconcile conflicts among a rapidly growing population and the need to conserve the habitat of a number of declining wildlife species in western Riverside County, California. Several important goals have been pursued by RCHCA member governments in their sponsorship of multiple-species habitat conservation plans (MSHCPs), including (1) establishing certainty and control over future uses of land; (2) eliminating project-by-project negotiations with federal and state wildlife agencies; (3) coordinating mitigation obligations under the Endangered Species Act, Clean Water Act, California Environmental Quality Act, and other federal and state laws; (4) reducing conflict and litigation resulting from land development activities; and (5) ensuring that wildlife conservation activities are conducted in a manner that permits local governments to perform those functions necessary to maintain public health, safety, and welfare. I also describe the emergence of strategies by local governments to achieve MSHCP goals, including (1) use of an inclusive planning process that seeks to build consensus among affected interests; (2) extensive involvement of federal and state wildlife agencies in the preparation of MSHCP documents; (3) management of public lands to support MSHCP conservation objectives; (4) encouragement of voluntary conservation by private property owners through incentive programs; and (5) active solicitation of federal and state funding for MSHCP implementation activities. PMID- 10801988 TI - A Crosswalk from the Endangered Species Act to the HCP Handbook and Real HCPs. AB - The U.S. Departments of Interior and Commerce published the HCP handbook in 1996 to guide the issuance of incidental take permits (ITPs). The HCP handbook lists six guidelines and provides many others throughout the text. However, the guidelines sometimes contradict the intent of the Endangered Species Act (ESA), and some are vague due to their use of improperly defined terms, such as ecosystem, region(al), net benefits, and habitat types. A proposed addendum provides little improvement to the HCP handbook. The guidelines in the HCP handbook could be prepared to better match those that scientists would expect from the language of the ESA. Three HCPs met only 3-10% of the 39 guidelines I would expect based on the ESA, and they met only 7-20% of the 30 guidelines in the HCP handbook. Based on a recent legal decision on the Alabama Beach Mouse HCP, ITP applicants are expected to follow the guidelines in the HCP handbook. The three HCPs I reviewed are vulnerable to similar legal decisions. However, many of the guidelines in the HCP handbook are inconsistent with the ESA. Therefore, I recommend that the HCP handbook be revised so that it is consistent with the ESA and with academic standards in the use of scientific methods. The guidelines need to be more explicit to be operationally consistent. Such a revision would provide ITP applicants and all the stakeholders with much more certainty in the outcome of the process. In lieu of a revised HCP handbook, I recommend that ITP applicants prepare their HCPs using the standards presented in Smallwood and others (Environmental Management, 1999, 24:421-435), thereby giving their HCPs a strong scientific foundation. PMID- 10801989 TI - The Selection and Design of Multiple-Species Habitat Preserves. AB - Multiple-species habitat conservation plans (MSHCPs) are designed to eliminate project-by-project review and minimize species-by-species conflicts; but these one-time, short-term processes invariably compress the divergent expectations of interest groups into an exercise driven by economic, amenity, and aesthetic values rather than scientific values. Participants may define an MSHCP as an exchange of habitat preserves for federal permits to take populations of endangered animals and plants, but the outcome is typically driven by overarching arguments over land development and suburban sprawl. Existing land uses also constrain the size, shape, and linkages among wildlife habitats, leading to a divergence of MSHCPs from the scientific preserve selection and design literature. Problems created by constraints to preserve configuration (e.g., land costs, fragmentation, pre-existing amounts of edge, lack of connectivity) must be resolved by long-term, post facto management. To date, estimates of preserve persistence have not been used in MSHCPs. Rather than focus on map-based exercises of preserve elements, it may be more productive to set goals for the persistence of species (states) and ecosystems (processes) within the preserves accepting that preserve configurations and arrays will be defined by the landscape and politics of suburban areas and that long-term management will provide the primary means of maintaining biodiversity along the wildland/urban interface. PMID- 10801990 TI - Problems in Extinction Model Selection and Parameter Estimation. AB - It is a vexing problem to achieve a consensus about the proper scientific way to assess population viability for habitat conservation plans. Rather than a hypothesis-testing approach, here it is proposed to select population models, estimate extinction parameters, and assess prediction uncertainty using a pragmatic, empirical Bayesian approach. The simplest usable models include the effects of population growth, r; carrying capacity, K; Allee threshold, N(A); and environmental stochasticity, v(r). Analytic predictions of expected extinction times are available for such models. Models that are more complex can be elaborated from this basis. Selection from a hierarchy of nesting population models can often be done through the evaluation of parameters. The estimation of the most important extinction parameters can be undertaken in a variety of ways. Time series can be analyzed to estimate r(d), v(r), rho, and K. Habitat models and individualistic population models may help estimate N(A) and K and demographic stochasticity. Fine-scale biogeography and climatological data may be useful in the estimation of a variety of parameters. Because it takes many years to estimate extinction parameters accurately for a given population of interest, the most efficient estimation procedures are desirable. I propose the use of prior information from an (as yet nonexistent) population biology database. The accumulation of local information through monitoring will improve our estimates allowing adaptive management. Uncertainty in the estimates will always remain, but it may be quantified by the posterior distributions. A crude example is discussed using treefrog population data. Although the motivations, beliefs, and biases of competing stakeholders will differ, a habitat conservation plan could accommodate this variation in the prior distributions. Field experience from monitoring will increasingly clear up any discrepancies between the opposing beliefs and the real ecosystem. As the world is an uncertain place and because there is no universal scientific method, there will always be controversy and surprises. The best we can do is (1) agree about our prior information, (2) agree about the strategy of model selection and parameter estimation, and (3) agree about our strategy for adaptive management. Perhaps the greatest impediment to such prior agreements for HCPs is the likely paranoia inspired by the use of unfamiliar statistical methodology. We need to train students of ecology in a more flexible and deeper understanding of statistics and philosophy of science. PMID- 10801991 TI - Conservation and Management for Multiple Species: Integrating Field Research and Modeling into Management Decisions. AB - Multiple-species reserves aim at supporting viable populations of selected species. Population viability analysis (PVA) is a group of methods for predicting such measures as extinction risk based on species-specific data. These methods include models that simulate the dynamics of a population or a metapopulation. A PVA model for the California gnatcatcher in Orange County was developed with landscape (GIS) data on the habitat characteristics and requirements and demographic data on population dynamics of the species. The potential applications of this model include sensitivity analysis that provides guidance for planning fieldwork, designing reserves, evaluating management options, and assessing human impact. The method can be extended to multiple species by combining habitat suitability maps for selected species with weights based on the threat faced by each species, and the contribution of habitat patches to the persistence of each species. These applications and extensions, together with the ability of the model to combine habitat and demographic data, make PVA a powerful tool for the design, conservation, and management of multiple species reserves. PMID- 10801992 TI - Ecological Restoration of Coastal Sage Scrub and Its Potential Role in Habitat Conservation Plans. AB - Extensive acreage loss of coastal sage scrub (CSS), isolation of surviving stands, and the federal listing of several animal species with obligate relationships to this plant community, particularly the threatened California gnatcatcher (Polioptila californica), have led to attempts to create CSS to mitigate habitat lost to urban development and other causes. Many of these creations lie within habitat conservation plan (HCP) sites, and they could play a more prominent role by being repositories for plants taken from a single site having site-specific genetics. Among others, one technique that increases initial resemblance to natural stands uses digitized, to-scale photography, which has been ground-truthed to verify vascular plant associations, which appear as mosaics on a landscape. A combination of placing patches of salvaged, mature canopy plants within larger matrices of imprinted or container plant plots appears to significantly enhance immediate use by CSS obligate bird species, accelerate "spread" or expansion of CSS, and can also introduce many epiphytic taxa that otherwise would be slow or unable to occupy developing CSS creations. Reptile, amphibian, butterfly, and rodent diversity in a salvaged canopy restoration case study at the University of California, Irvine, showed CSS species foraging and inhabiting transplanted canopy patches. Using restoration techniques to expand existing CSS stands has more promise than creating isolated patches, and the creation of canopies resembling CSS mid-fire cycle stands is now common. Gnatcatchers and other birds use restorations for foraging and occasional nesting, and in some cases created stands along "biological corridors" appear to be useful to bird movement. Patches of transplanted sage scrub shrubs along habitat edges appear to break up linear edge effects. There are no data on which long-term survival, succession, or postfire behavior can be predicted for CSS restoration sites, and postfire community changes are not part of either mitigation or restoration planning at present. Long-term planning including burning is needed so that a fire-adapted habitat will develop. Restoration is important in retaining genetic resources, for ameliorating edge effects, as habitat extenders in buffer zones around HCP sites, and by providing areas into which natural stands can expand. PMID- 10801993 TI - Arthropods and Multispecies Habitat Conservation Plans: Are We Missing Something? AB - Arthropods constitute well over one-half of the species of higher life on the planet and are the dominant terrestrial life form on the planet. Unfortunately, very little is known about most arthropod species. There are an estimated 163,487 species of insects in North America, of which only 66% are taxonomically known. Similarly, there are an estimated 35,514 species of North American arachnids, of which only 9316 are described; over 73% have yet to be discovered and described. Without the basic taxonomic and life history knowledge for most of the terrestrial species (i.e., arthropods) of North American ecosystems, land managers are faced with the challenge of developing, selecting, and managing biotic reserves and habitat conservation plans for which they know very little about the majority of organisms found within such reserves or covered by such plans. With respect to arthropods, this challenge includes taking into account poorly described species being used as political tools to stop development (as opposed to actually protecting a truly endangered species), thus confounding the habitat conservation planning process and ensuring that "surprises" in the form of new listings will occur within any multispecies habitat plan. Finally, using various scenarios and assumptions, estimates of the true number of endangered insects and arachnids are provided to illustrate the fact that the suspected number of threatened, endangered, and extinct species is probably low by at least an order of magnitude. PMID- 10801994 TI - Effect of pravastatin on plasma removal of a chylomicron-like emulsion in men with coronary artery disease. AB - The speed of the plasma removal of chylomicrons, the lipoproteins that carry dietary lipids absorbed in the intestine, may influence atherogenesis. Thus, the effects of a 30-day pravastatin or placebo treatment on the plasma kinetics of chylomicron-like emulsions were evaluated in 25 patients with coronary artery disease who were not hypertriglyceridemic in a randomized, single-blinded study. Eleven patients (53 +/- 4 years, 10 men) received pravastatin 40 mg/day and 14 received placebo (52 +/- 3 years, 13 men). Emulsions labeled with triolein ((3)H TO) and cholesteryl oleate ((14)C-CO) to assess lipolysis and clearance of chylomicron and remnants, respectively, were injected intravenously in a bolus after a 12-hour fast. Blood samples were collected during 60 minutes to determine radio isotope decaying curves and fractional catabolic rates. Subjects were studied at baseline and after the treatment period. Compared with placebo (data expressed as mean +/- SEM), pravastatin treatment increased the (14)C-CO fractional catabolic rates (70 +/- 45% vs 18 +/- 10%, p = 0.01), reduced total cholesterol (-21 +/- 3% vs -3 +/- 2% p = 0.0001), low-density lipoprotein (LDL) cholesterol (-25 +/- 5% vs 4 +/- 6%, p = 0.0001), and apolipoprotein B levels ( 22 +/- 3% vs -7 +/- 3% p = 0.01). (3)H-TO fractional catabolic rates, plasma triglycerides, very-low-density lipoprotein (VLDL) cholesterol and high-density lipoprotein (HDL) cholesterol variations did not differ between the groups. The fractional catabolic rate of (14)C-CO was inversely correlated with plasma apolipoprotein B levels (r = -0.7, p = 0.04). This suggests that besides reducing LDL cholesterol, pravastatin also increases chylomicron remnant clearance, with possible antiatherogenic implications. PMID- 10801995 TI - Antithrombotic effects of abciximab. AB - The observation that platelet-platelet interaction and thrombosis are ultimately regulated by the glycoprotein (GP) IIb/IIIa receptor complex, triggered the development of agents capable of interfering with this platelet receptor complex. Several large clinical trials have demonstrated the effectiveness of this class of agents. The first of these agents to show beneficial effects after coronary interventions was the mouse/human chimeric Fab fragment antibody c7E3 (abciximab; ReoPro). This study analyzes whether the addition of heparin to the GP IIb/IIIa antagonist abciximab would enhance the antithrombotic effect. Blood drawn directly from patients on aspirin who underwent interventional procedures perfused an ex vivo perfusion chamber containing a severely injured arterial wall at local rheologic conditions of a mildly stenosed coronary artery. Blood was perfused directly from patients at baseline and following administration of heparin, abciximab, or both. The antithrombotic effects of the 3 treatments were assessed by reduction of the thrombus formation on the perfused specimens. Thrombus formation at baseline was not significantly modified by the administration of heparin (13,897 +/- 1,316 vs 11,917 +/- 1,519 microm(2)). Abciximab produced a 58% reduction in thrombus formation (11,631 +/- 861 vs 4, 925 +/- 585 microm(2); p <0.001). The addition of heparin to abciximab did not further reduce thrombus area versus abciximab alone (5,651 +/- 581 vs 4,925 +/- 585 microm(2)). Thus, our data show that abciximab dramatically decreases mural thrombus formation and that combining heparin with abciximab did not add any additional antithrombotic effect to abciximab alone. PMID- 10801996 TI - Usefulness of carvedilol in unstable angina pectoris. AB - The safety and efficacy of adding oral carvedilol (25 mg twice daily) to standardized treatment of unstable angina was assessed in a multicenter, randomized, double-blind, placebo- controlled trial on 116 patients with acute unstable angina. Patients were monitored in an intensive care unit and underwent 48-hour Holter monitoring to assess transient ischemia. Carvedilol as adjunctive therapy resulted in a significant reduction of median heart rate (65 vs 75 beats/min, p <0.05), mean systolic blood pressure (133 vs 130 mm Hg, p <0.05), and mean rate-pressure product (8,337 vs 10,042, p <0.05). Carvedilol reduced the ischemic burden during 48 hours of treatment by 75% (49 vs 204 minutes), including a 36% reduction of patients with ischemic episodes (p <0.05), a 66% reduction of the mean number of ischemic episodes (8 vs 24, p <0.05), and a 76% reduction in the mean duration of ischemic episodes (50 vs 205 minutes, p <0.05). Side effects occurred in 8 of 59 patients (13.6%) in the carvedilol group and in 5 of 54 patients (8.8%) given placebo. Although not significant, the early onset of maximal blood pressure reduction and the delayed effect on heart rate were closely correlated to drug-induced hypotension and bradycardia in the carvedilol group. Thus, carvedilol as an adjunctive to standardized treatment effectively reduces heart rate and blood pressure, and thus the ischemic burden in patients with unstable angina pectoris, but requires close monitoring of patients at risk for bradycardia or hypotension. PMID- 10801997 TI - A simplified lesion classification for predicting success and complications of coronary angioplasty. Registry Committee of the Society for Cardiac Angiography and Intervention. AB - In 1988, the American College of Cardiology/American Heart Association (ACC/AHA) Task Force on Assessment of Diagnostic and Therapeutic Cardiovascular Procedures presented a classification of coronary lesions utilizing 26 lesion features to predict the success and complications of balloon angioplasty. Using data from the Registry of the Society for Cardiac Angiography and Interventions (SCAI) we evaluated the ability of this classification to predict success and complications. Lesion success, death in hospital, emergency cardiac bypass surgery, and major adverse events were evaluated in 41,071 patients who underwent single-vessel angioplasty from January 1993 to June 1996. Logistic models using the ACC/AHA lesion classification, vessel patency, or both, were compared. A new classification based on the interaction of the ACC/AHA classification plus lesion patency was compared with the existing ACC/AHA classification. Vessel patency, added to the ACC/AHA classification, improved prediction of lesion success (p 90 ms was 2.8-fold higher than those with 01 dispersion 90 ms (95% confidence intervals [CI] 1.2 to 6.4). Similarly, the death rate in patients with QRS dispersion >46 ms was 3.9-fold higher than in those with QRS dispersion 46 ms (95% Cl 1.6 to 9.5). These findings suggest that QT and QRS dispersion are useful predictors of mortality in patients with advanced CHF. ?2000 by Excerpta Medica, Inc. PMID- 10802004 TI - von Willebrand factor, fibrinogen, and soluble P-selectin levels after mitral valve replacement versus mitral valve repair. AB - Patients with mitral valve disease undergoing surgery are at an increased risk of thromboembolism. We hypothesized that this may be due in part to abnormalities in platelet activation, endothelial damage or dysfunction, and plasma fibrinogen in such patients. To test this hypothesis, we measured indexes of platelet activation (soluble P-selectin), endothelial damage or dysfunction (von Willebrand factor [vWf], enzyme-linked immunosorbent assay) and fibrinogen (modified Clauss) in 56 consecutive patients (35 women, mean age 65 years) admitted for isolated mitral valve repair (n = 39) or replacement (using mechanical implants, n = 17). Samples were taken from a peripheral vein before and at 3 months after valve surgery. Baseline results were compared with 56 healthy age- and sex-matched controls. Compared with controls, patients with mitral valve disease had higher levels of vWf (mean +/- SD 132 +/- 28 vs 101 +/- 35 IU/dl; p <0.001), but there were no significant differences in mean fibrinogen (p = 0.418) or soluble P-selectin (p = 0.855) levels between cases and controls. There was a significant increase in plasma vWf after mitral valve replacement: 142 +/- 25 IU/dl preoperatively, increasing to 161 +/- 33 IU/dl at 3 months after surgery (p = 0.0261). However, there were no significant changes in plasma fibrinogen (p = 0.306) or soluble P-selectin levels (p = 0.191). Patients undergoing mitral valve repair did not have any significant changes in mean vWf (p = 0.25), soluble P-selectin (p = 0.77), or fibrinogen (p = 0.22). There was a significant negative correlation (Spearman, r = -0.4, p = 0.003) in postoperative plasma vWf levels and the size of valve prosthesis used. Thus, patients with mitral valve disease have increased plasma vWf levels when compared with healthy controls, suggesting endothelial damage or dysfunction, with a further increase in levels after mitral valve replacement. Conversely, patients undergoing mitral valve repair do not demonstrate any significant changes in fibrinogen, or indexes of endothelial dysfunction or platelet activation. PMID- 10802005 TI - Ventilatory response to exercise in patients with major aortopulmonary collateral arteries after definitive surgery. AB - Patients with pulmonary atresia and major aortopulmonary collateral arteries (MAPCAs) may be at risk for both ventilatory impairment and abnormal pulmonary circulation after definitive surgery. We measured the ventilatory response to exercise in 16 patients with MAPCAs after definitive surgery (group A) and compared the results with those in 16 patients with tetralogy of Fallot with pulmonary atresia and without MAPCAs after definitive operation (group B), with 24 patients with tetralogy of Fallot after one-stage repair without previous palliation (group C), and with 48 healthy subjects (group D). Pulmonary function and treadmill exercise tests were performed. Arterial blood gases were also analyzed and the dead space to tidal volume ratio calculated. In group A, the vital capacity, diffusion capacity, and peak oxygen uptake were lowest (p <0.001), whereas the ventilatory equivalent for carbon dioxide was highest and its value at peak exercise correlated with age at time of surgery (r = 0.73, p <0.002). The arterial oxygen tension decreased progressively in group A and its value at peak exercise inversely correlated with the mean pulmonary artery pressure in all patients (r = -0.75, p <0.001). The arterial carbon dioxide tension decreased significantly at peak exercise in controls but showed no change in group A. The dead space to tidal volume ratio decreased during exercise in patients without MAPCAs and in controls but increased in group A, and the dead space to tidal volume ratio at peak exercise was inversely correlated with vital capacity in all patients (r = -0.77, p <0.001). Diffusion capacity independently predicted arterial carbon dioxide tension and dead space ventilation during exercise. Marked restrictive ventilatory impairment with low diffusion capacity along with a pulmonary obstructive change contributed to the abnormal pulmonary gas exchange during exercise in group A. Earlier repair of MAPCAs may prevent the progression of the impaired ventilatory response to exercise in these patients. PMID- 10802006 TI - World experience of percutaneous ultrasound-guided balloon valvuloplasty in human fetuses with severe aortic valve obstruction. AB - Prenatal alleviation of severe fetal aortic valve obstructions by percutaneous ultrasound-guided balloon valvuloplasty has been performed to improve the fate of affected fetuses. The purpose of this study was to analyze the current world experience of these procedures in human fetuses. Data from 12 human fetuses were available for analysis. The mean gestational age at intervention was 29.2 weeks (range 27 to 33). The mean time period between initial presentation and intervention was 3.3 weeks (range 3 days to 9 weeks). Technically successful balloon valvuloplasties were achieved in 7 fetuses, none of whom had an atretic valve. Only 1 of these fetuses remains alive today. Of the 5 remaining technical failures, 1 patient with severe aortic stenosis underwent successful postnatal intervention and remains alive. Six patients who survived prenatal intervention died from cardiac dysfunction or at surgery in the first days or weeks after delivery. Four fetuses died early within 24 hours after the procedure, 1 from a bleeding complication, 2 from persistent bradycardias, and 1 at valvotomy after emergency delivery. Thus, the early clinical experience of percutaneous ultrasound-guided fetal balloon valvuloplasty in human fetuses with severe aortic valve obstruction has been poor due to selection of severe cases, technical problems during the procedure, and high postnatal operative mortality in fetuses who survived gestation. Improved patient selection and technical modifications in interventional methods may hold promise to improve outcome in future cases. PMID- 10802007 TI - Improved reliability for echocardiographic measurement of left ventricular volume using harmonic power imaging mode combined with contrast agent. AB - Harmonic power imaging (HPI) is a new echocardiographic modality that enhances the detection of contrast agents in the left ventricle. The endocardium can be delineated by conventional echocardiography using ultrasound contrast agents, although the images tend to be faint. The present study was designed to assess left ventricular volume using HPI after intravenous injection of the contrast agent Levovist (Schering SA, Berlin, Germany) in 25 unselected patients. End diastolic volume, end-systolic volume, and ejection fraction were determined for each patient with angiography and with 4 different ultrasound modalities: (1) conventional mode without contrast, (2) contrast conventional mode, (3) contrast harmonic intermittent imaging mode, and (4) contrast triggered HPI. The use of HPI improved correlations between the echographic and angiographic measurements for all parameters as well as precision and bias determined by Bland and Altman analysis. The relative errors for interobserver variability were also lower with HPI. This study demonstrates that echocardiographic determination of left ventricular volumes and ejection fraction is more accurate and reproducible using HPI combined with Levovist. PMID- 10802008 TI - Omega-3 fatty acids: time for clinical implementation? PMID- 10802009 TI - Outcome 14 to 18 years after percutaneous transluminal coronary angioplasty. PMID- 10802010 TI - Resolution of coronary thrombus with rescue stenting. PMID- 10802011 TI - Cost-effective analysis of primary infarct-artery stenting versus optimal primary angioplasty (the Florence Randomized Elective Stenting in Acute Coronary Occlusions [FRESCO] trial). PMID- 10802012 TI - Comparison of nitroglycerin magnetic resonance imaging with dobutamine echocardiography for predicting recovery of function after revascularization. PMID- 10802013 TI - Association between height and coronary artery disease in black men and women. PMID- 10802014 TI - Autonomic modulation and atrial fibrillation in the Wolff-Parkinson-White syndrome. PMID- 10802015 TI - Risk factors associated with development of atrial fibrillation early after coronary artery bypass grafting. PMID- 10802016 TI - Usefulness of intravenous granisetron for prevention of neurally mediated hypotension upon head upright tilt testing. PMID- 10802017 TI - Possible role of increased blood viscosity in the hemodynamics of systemic hypertension. PMID- 10802018 TI - Mitral valve prolapse in healthy relatives of patients with familial Pseudoxanthoma elasticum. PMID- 10802019 TI - Change in mitral regurgitation severity after aortic valve replacement for aortic stenosis. PMID- 10802020 TI - Prevalence of circulating Trypanosoma cruzi detected by polymerase chain reaction in patients with Chagas' cardiomyopathy. PMID- 10802021 TI - Effects of short-term estrogen treatment on the neointimal response to balloon injury of rat carotid artery. PMID- 10802022 TI - Life-threatening ventricular arrhythmias associated with giant cell myocarditis (possibly sarcoidosis). PMID- 10802023 TI - Distribution of NADPH-diaphorase cells in visual and somatosensory cortex in four mammalian species. AB - The distribution of the well-labeled nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) Type I neurons was evaluated in the isocortex of four mammalian species: the Didelphis opossum, the Monodelphis opossum, the rat and the marmoset. In Didelphis opossum, laminar distribution was examined in tangential and non-tangential sections. The density increases from superficial to deep layers of the gray matter. In rats' tangential sections, infragranular and supragranular layers have higher density than layer IV. Cell density measurements in the visual and the somatosensory cortices were compared in tangential sections from flattened hemispheres of the four species. Somatosensory areas were identified histochemically in rat (barrel fields) and marmoset (S1 and S2/PV). In the opossums, areas S1 and S2/PV were identified by multiunit recording. Except in the rat, primary visual cortex (V1) was labeled histochemically by NADPHd and/or cytochrome oxidase. In the four species, cell density in somatosensory cortex was significantly higher than in visual cortex. Taken together these results demonstrate that NADPHd Type I neurons are not homogeneously distributed in the isocortex of these mammals. In conclusion, the tangential distribution of Type I neurons in the sensory areas examined, but not its laminar distribution, was similar in the four species. Given that rats, marmosets and opossums are distantly related species, and that the latter are considered to have more 'generalized' brains, it is conceivable that this pattern of tangential distribution of Type I neurons is a general feature of mammalian isocortex. PMID- 10802024 TI - Ultrastructural localization of the serotonin 2A receptor in dopaminergic neurons in the ventral tegmental area. AB - Serotonin (5-hydroxytryptamine, 5-HT) 2A receptor antagonists are clinically effective antipsychotics that may differentially target mesocortical and mesolimbic dopaminergic neurons having partially segregated distribution in the parabrachial (PB) and paranigral (PN) ventral tegmental area (VTA). We examined the ultrastructural immunocytochemical localization of the 5-HT2A receptor in these subdivisions of rat VTA, to determine (1) the functional sites for receptor activation, and (2) cellular associations between the receptor and dopaminergic neurons identified by their content of tyrosine hydroxylase (TH). The mean area density of neuronal profiles containing 5-HT2A receptor labeling was not significantly different in the PB and PN VTA. In each region approximately 44% of the 5-HT2A labeled profiles were dendrites while the remainder were mainly axons. Dendritic 5-HT2A-immunoreactivity was often localized to membranous cytoplasmic organelles resembling smooth endoplasmic reticulum, and to more rarely to segments of the plasma membrane beneath contacts from unlabeled axon terminals. 5 HT2A labeling was also seen within the cytoplasm of a few axon initial segments and many small unmyelinated axons. Approximately 40% of the 5-HT2A-labeled dendritic profiles contained TH in either PB or PN VTA. Our results suggest that 5-HT2A receptors in VTA are largely cytoplasmic and play an equally important role in modulating dopaminergic neurons in PB and PN VTA. These results also implicate 5-HT2A receptors in the postsynaptic activation of non-dopaminergic neurons and possibly the presynaptic release from terminals of axons originating in, or passing through, these regions. PMID- 10802025 TI - Cerebellar CB(1) receptor mediation of Delta(9)-THC-induced motor incoordination and its potentiation by ethanol and modulation by the cerebellar adenosinergic A(1) receptor in the mouse. AB - The effect of intracerebellar microinfusion of antisense oligodeoxynucleotide to Delta(9)-tetrahydrocannabinol (Delta(9)-THC) and other naturally occurring cannabinoid receptor (CB(1)) mRNA on Delta(9)-THC-induced motor impairment was investigated in mice. Delta(9)-THC (15-30 microgram/microliter intracerebellar) resulted in a significant motor impairment in a dose-related manner. The intracerebellar pretreatment with antisense oligodeoxynucleotide (3.0 microgram/100 nl/12 h; six administrations/mouse) virtually abolished Delta(9) THC (15 and 25 microgram/1 microliter intracerebellar)-induced motor impairment. However, intracerebellar pretreatment with the mismatched oligodeoxynucleotide in exactly the same manner as the antisense was completely ineffective in altering the Delta(9)-THC-induced motor impairment. These results strongly suggest the involvement of CB(1) receptor in the expression of Delta(9)-THC-induced motor impairment. The intracerebellar microinfusion of adenosine A(1)-selective agonist, N(6)-cyclohexyladenosine (CHA) (4 ng/100 nl) significantly enhanced Delta(9)-THC-induced motor impairment, suggesting a cerebellar A(1) adenosinergic modulation of motor impairment. A pretreatment with the antisense and the mismatched oligodeoxynucleotide also markedly attenuated and did not alter, respectively, the cerebellar A(1) adenosinergic modulation (enhancement) of Delta(9)-THC-induced motor impairment. There was no change in the normal motor coordination due to intracerebellar pretreatment with antisense and its mismatch, in the presence as well as absence of intracerebellar CHA indicating the selectivity of interactions with Delta(9)-THC. The Delta(9)-THC-induced motor incoordination was also significantly enhanced dose-dependently by systemic (i.p.) ethanol administration suggesting behavioral synergism between the two psychoactive drugs. Pretreatment (intracerebellar) with pertussis toxin (PTX) markedly attenuated Delta(9)-THC- and Delta(9)-THC+CHA-induced motor incoordination suggesting coupling of CB(1) receptor to PTX-sensitive G-protein (G(i)/G(o)). These data suggested co-modulation by cerebellar cannabinoid and adenosine system of Delta(9)-THC-induced motor impairment. Conversely, the results in the present study also suggested co-modulation by cerebellar adenosine A(1) and CB(1) receptors of ethanol-induced motor impairment, thereby indicating a possible common signal transduction pathway in the expression of motor impairment produced by Delta(9)-THC as well as ethanol. PMID- 10802026 TI - Effect of thyroid hormone deficiency on developmental expression of goalpha gene in the brain of neonatal rats by competitive RT-PCR and in situ hybridization histochemistry. AB - Goalpha is a guanine nucloetide-binding regulatory protein alpha subunit which is mainly distributed in the central nervous system, but it has not previously been reported how it is regulated by thyroid hormone in the brain of neonatal rat at transcriptional levels. In this report, we used quantitative competitive reverse transcriptional PCR to quantify the effects of TH deficiency on Goalpha gene expression in the brain of neonatal rat at mRNA levels. It was found that Goalpha mRNA levels in the brain of 14-day-old rats significantly increased over 3-fold after induction of perinatal hypothyroidism, and declined markedly after treatment of thyroxine replacement. In situ hybridization histochemistry was further employed to observe the time-course and spatial expression of Goalpha gene in the brain of neonatal rats affected by thyroid hormone deficiency during the developmental period. The data showed that perinatal hypothyroidism can enhance Goalpha mRNA levels in the temporal cortex, sensorimotor cortex, piriform cortex, amygdala, hippocampal CA1-4 subfields, dentate gyrus, arcuate nucleus (AR) and ventromedial hypothalamic nucleus (VMH) of hypothalamus, but not in the striate cortex, cingulate cortex, claustrum, caudate/putamen and thalamus in the brain of rat at 7-21 days post-partum. The results suggest that up-regulation of Goalpha gene expression may be one kind of common mechanism responsible for neurological deficits in some brain areas arising from thyroid hormone deficiency in the critical periods of neonatal rats. PMID- 10802027 TI - Regional cerebral blood flow responses to variable frequency whisker stimulation: an autoradiographic analysis. AB - Activation of the rat primary somatosensory barrel field (S1BF) is a commonly used model to study the mechanisms of evoked coupled cortical blood flow changes. However, the relationship between these blood flow changes and variable whisker movement has not been completely characterized. We have previously shown that in urethane anesthetized rats, the magnitude of laser-Doppler measured cortical blood flow changes increase linearly with the frequency of full pad whisker movement over the physiological range of 1.5 to 10.5 s. To further test the hypothesis that local cortical blood flow increases with frequency of whisker movement and underlying neuronal activity, regional cerebral blood flow (rCBF) was determined autoradiographically in seven urethane anesthetized SD rats. Selected rows of whiskers (rows C, D, E) were stimulated at 3 s on the right side of the rat's face and simultaneously at 10 s on the left side for 2 min prior to radioactive tracer administration. Subregions of somatosensory cortex were identified with the aid of thionin and cytochrome oxidase stained sections. Mean rCBF (ml/100 g/min) for S1BF were: S1BF [0 s] left cortex, 146+/-13; S1BF [0 s] right cortex, 158+/-15; S1BF[3 s], 160+/-13; S1BF [10 s] 178+/-14. In both stimulated and nonstimulated regions, the profile of blood flow increased across cortex laminae, peaking in layer IV and decreasing through deeper layers. Maximal blood flow increases elicited by whisker movement occurred in cortical layers I IV. These data support the hypothesis that whisker movement elicited rCBF changes are input frequency dependent and are most pronounced in cortical layers I though IV. These data provide a strong framework in which to study the mechanisms of neuronal activity-blood flow coupling. PMID- 10802028 TI - Effects of TNFalpha on immature and mature oligodendrocytes and their progenitors in vitro. AB - Tumor necrosis factor alpha (TNFalpha) appears to take part in the pathogenesis of multiple sclerosis and to contribute to the degeneration of oligodendrocytes as well as neurons. TNFalpha is produced by microglia and astrocytes, which also produce hormones and cytokines that influence its biological activity. Thus, in mixed cultures the effects of exogenous TNFalpha might be modified by products of astrocytes and microglia. The effects of TNFalpha in oligodendrocyte-enriched cultures are reported below. We prepared the cultures by shaking oligodendrocytes off primary mixed glial-cell cultures from brains of 2-day-old rats at 7 days in vitro and plating them (0 days post-shake, DPS). Platelet-derived growth factor and fibroblast growth factor were included in the media at 1-5 DPS in order to encourage proliferation. At 2 DPS media were added with no TNFalpha (controls) or 1000, 2000 or 5000 U/ml of TNFalpha, and at 5 DPS media were replaced with fresh serum-free media. Cultures were fixed with 4% paraformaldehyde at 5, 7, 9 and 12 DPS and immunostained. Oligodendrocyte progenitors were not reduced in numbers immediately after the incubation with TNFalpha (i. e. at 5 DPS). However, after an additional 4 days in culture fewer progenitors remained in the cultures that had been treated with TNFalpha than in the untreated cultures. In the absence of the growth factors there were fewer progenitors, but their numbers also were reduced by TNFalpha. Maturation to the myelin basic protein (MBP)-positive stage was inhibited by about 36% at 9 DPS by 1000-2000 U/ml of TNFalpha, while numbers of O4+/MBP- precursors were unaffected. It is interesting that the steady-state number of O4-positive precursors was unchanged by TNFalpha at 9 DPS, when there were reductions in the numbers of A2B5-positive progenitors and MBP-positive mature oligodendrocytes. That observation suggests that the rates of proliferation, death and maturation are controlled by multiple factors, with a particularly vulnerable time at the maturation to the MBP-positive stage. At 5000 U/ml TNFalpha the specific effect on maturation was overtaken cytotoxicity. These data and a summary of the literature suggest that inhibition of MBP expression is sensitive to lower TNFalpha concentrations and incubation times than is cell survival. Specific effects on numbers of MBP-positive cells, morphology and MBP expression occur at 1000-2000 U/ml for 48-72 h or at up to 10000 U/ml forapelin-17>apelin-13>apelin-12. PMID- 10802051 TI - Accumulation of 210 kDa microtubule-interacting protein in differentiating P19 embryonal carcinoma cells. AB - The MA-01 antigen, a thermolabile 210 kDa microtubule-interacting protein, is present in P19 embryonal carcinoma cells on microtubular structures as well as in cytosol. After aggregation of the cells and subsequent incubation with all-trans retinoic acid (RA), the level of MA-01 expression increased approximately 10 times during 15 days. The increase started after 2 days of incubation with RA and preceded the appearance of neuron-specific tubulin betaIII, MAP2C and neurofilaments. Such elevated expression of MA-01 antigen was not detected in P19 cells treated with dimethylsulfoxide. These data indicate that enhanced expression of MA-01 antigen is one of the earliest events occurring in P19 cells during neuronal differentiation. PMID- 10802052 TI - Antioxidant alpha-tocopherol ameliorates glycemic control of GK rats, a model of type 2 diabetes. AB - We have shown recently that oxidative stress by chronic hyperglycemia damages the pancreatic beta-cells of GK rats, a model of non-obese type 2 diabetes, which may worsen diabetic condition and suggested the administration of antioxidants as a supportive therapy. To determine if natural antioxidant alpha-tocopherol (vitamin E) has beneficial effects on the glycemic control of type 2 diabetes, GK rats were fed a diet containing 0, 20 or 500 mg/kg diet alpha-tocopherol. Intraperitoneal glucose tolerance test revealed a significant increment of insulin secretion at 30 min and a significant decrement of blood glucose levels at 30 and 120 min after glucose loading in the GK rats fed with high alpha tocopherol diet. The levels of glycated hemoglobin A1c, an indicator of glycemic control, were also reduced. Vitamin E supplementation clearly ameliorated diabetic control of GK rats, suggesting the importance of not only dietary supplementation of natural antioxidants but also other antioxidative intervention as a supportive therapy of type 2 diabetic patients. PMID- 10802053 TI - Resistance to CD95/Fas-induced and ceramide-mediated apoptosis of human melanoma cells is caused by a defective mitochondrial cytochrome c release. AB - Intracellular CD95/Fas-signaling pathways have not been investigated in melanoma yet. Two different CD95 receptor-induced apoptotic pathways are presently known in other cell types: (i) direct activation of caspase-8 and (ii) induction of ceramide-mediated mitochondrial activation, both leading to subsequent caspase-3 activation. In the present study, five of 11 melanoma cell populations were shown to release cytochrome c from mitochondria, which activates caspase-3 and finally results in DNA fragmentation upon treatment with the agonistic monoclonal antibody CH-11. In contrast, this apoptotic pathway was not activated in the remaining six melanoma cell populations. Interestingly, the susceptibility of melanoma cells to CD95L/FasL-triggered cell death was clearly correlated with N acetylsphingosine-mediated apoptosis. Our results are in line with a defect upstream of mitochondrial cytochrome c release in resistant cells. PMID- 10802054 TI - alpha(1)-antitrypsin can increase insulin-induced mitogenesis in various fibroblast and epithelial cell lines. AB - alpha(1)-Antitrypsin (AT), the archetypal member of the superfamily of serine proteinase inhibitors, inhibits leukocyte elastase activity and thereby can prevent lung damage. Here we show that in fibroblasts from human fetal lung and mouse embryo as well as in certain epithelial cells AT can also enhance the stimulatory effects of insulin on DNA synthesis and cell proliferation. Warming of AT at a moderate (41 degrees C) temperature for a longer time (21 h) or at a higher (65 degrees C) temperature for 30 min before treatment increased its stimulatory effects on both DNA synthesis and activating phosphorylation of p42/p44 mitogen-activated protein kinases. The results suggest that AT may promote regeneration of damaged tissues under pathophysiological conditions which are associated with fever. PMID- 10802055 TI - Identification of residues in the TPR domain of Ssn6 responsible for interaction with the Tup1 protein. AB - Ssn6, a yeast protein that comprises 10 tandem tetratricopeptide repeat (TPR) motifs, associates with Tup1 repressor protein and acts as a transcriptional corepressor. In this report we identify point mutations in the TPR1 of Ssn6 that disrupt Tup1 interaction. Furthermore, we construct a 3D model of the TPR domain of Ssn6, which is responsible for Tup1 binding, based on the known structure of protein phosphatase 5. According to this model all selected mutations reduce the ability of Ssn6 to interact with Tup1 by affecting the structural integrity of TPR1 and/or the correct spatial arrangement of TPR1 relative to TPR2 and TPR3. PMID- 10802056 TI - The inhibitory effect of the 5' untranslated region of muscle acylphosphatase mRNA on protein expression is relieved during cell differentiation. AB - Previous experiments suggested that the upstream AUG triplet present in the 5' untranslated region (UTR) of muscle acylphosphatase mRNA is involved in the regulation of protein expression. In this paper, we study the involvement of the 5'UTR secondary structure and upstream peptide on mRNA stability and protein translation. Our data, obtained using deletion and frame-shift mutants, demonstrate that the 5'UTR controls protein expression regulating translation together with mRNA stability. Furthermore, we demonstrate that the inhibitory effect of the 5'UTR of muscle acylphosphatase is relieved during the differentiation process in agreement with previous data reporting an increase of acylphosphatase content during cell differentiation. Finally, UV cross-linking experiments show that specific mRNA-binding proteins are associated with the 5'UTR of the muscle acylphosphatase mRNA. PMID- 10802058 TI - Binding, annealing and strand exchange between oligonucleotides in different sites of the RecA protein filament. AB - The efficiency of single-stranded (ss) oligonucleotides binding at the secondary site of the RecA protein filament is demonstrated to depend on the strandedness of DNA bound at the primary site. When the primary site is occupied by a ss oligonucleotide, the binding of another ss-oligonucleotide at the secondary site is characterized by higher affinity and a lower rate of dissociation than is the case when the primary site is occupied by a double-stranded oligonucleotide. In contrast to a DNA strand exchange reaction suppressed by a heterologous oligonucleotide bound at the secondary site of the RecA filament, the occupation of the secondary site by a heterologous oligonucleotide does not prevent renaturation between the oligonucleotides bound at the primary site and complementary oligonucleotides from solution demonstrating that the binding of a DNA strand in the secondary site is not a necessary intermediate step in RecA promoted DNA renaturation. PMID- 10802057 TI - ENO1 gene product binds to the c-myc promoter and acts as a transcriptional repressor: relationship with Myc promoter-binding protein 1 (MBP-1). AB - The Myc promoter-binding protein-1 (MBP-1) is a 37-38 kDa protein that binds to the c-myc P2 promoter and negatively regulates transcription of the protooncogene. MBP-1 cDNA shares 97% similarity with the cDNA encoding the glycolytic enzyme alpha-enolase and both genes have been mapped to the same region of human chromosome 1, suggesting the hypothesis that the two proteins might be encoded by the same gene. We show here data indicating that a 37 kDa protein is alternatively translated from the full-length alpha-enolase mRNA. This shorter form of alpha-enolase is able to bind the MBP-1 consensus sequence and to downregulate expression of a luciferase reporter gene under the control of the c myc P2 promoter. Furthermore, using alpha-enolase/green fluorescent protein chimeras in transfection experiments we show that, while the 48 kDa alpha-enolase mainly has a cytoplasmic localization, the 37 kDa alpha-enolase is preferentially localized in the cell nuclei. The finding that a transcriptional repressor of the c-myc oncogene is an alternatively translated product of the ENO1 gene, which maps to a region of human chromosome 1 frequently deleted in human cancers, makes ENO1 a potential candidate for tumor suppressor. PMID- 10802060 TI - Adenylylsulfate reductases from archaea and bacteria are 1:1 alphabeta heterodimeric iron-sulfur flavoenzymes--high similarity of molecular properties emphasizes their central role in sulfur metabolism. AB - Highly active adenylylsulfate (APS) reductase was isolated under N(2)/H(2) from sulfate-reducing and sulfide-oxidizing bacteria and archaea. It was a 1:1 alphabeta-heterodimer of molecular mass approximately 95 kDa, and two subunits (alpha approximately 75, beta approximately 20 kDa). The specific activity was 11 14 micromol (min mg)(-1); cofactor analysis revealed 0.96+/-0.05 FAD, 7.5+/-0.1 Fe and 7.9+/-0.25 S(2-). The photochemically reduced enzyme had a multiline EPR spectrum resulting from two interacting [4Fe-4S] centers. The properties of the different APS reductases were remarkably similar, although the enzyme is involved in different metabolic pathways and was isolated from phylogenetically far separated organisms. A structural model is proposed, with FAD bound to the alpha subunit, and two [4Fe-4S] centers located in close proximity on the beta-subunit. PMID- 10802059 TI - Detection of superoxide anion using an isotopically labeled nitrone spin trap: potential biological applications. AB - We describe the synthesis and biological applications of a novel nitrogen-15 labeled nitrone spin trap, 5-ethoxycarbonyl-5-methyl-1-pyrroline N-oxide ([(15)N]EMPO) for detecting superoxide anion. Superoxide anion generated in xanthine/xanthine oxidase (100 nM min(-1)) and NADPH/calcium-calmodulin/nitric oxide synthase systems was readily detected using EMPO, a nitrone analog of 5,5' dimethyl-1-pyrroline N-oxide (DMPO). Unlike DMPO-superoxide adduct (DMPO-OOH), the superoxide adduct of EMPO (EMPO-OOH) does not spontaneously decay to the corresponding hydroxyl adduct, making spectral interpretation less confounding. Although the superoxide adduct of 5-(diethoxyphosphoryl)-5-methyl-pyrroline N oxide is more persistent than EMPO-OOH, the electron spin resonance spectra of [(14)N]EMPO-OOH and [(15)N]EMPO-OOH are less complex and easier to interpret. Potential uses of [(15)N]EMPO in elucidating the mechanism of superoxide formation from nitric oxide synthases, and in ischemia/reperfusion injury are discussed. PMID- 10802061 TI - Cd(II), Pb(II) and Zn(II) ions regulate expression of the metal-transporting P type ATPase ZntA in Escherichia coli. AB - ZntA is a cation-translocating ATPase which exports from Escherichia coli Cd(II) and Pb(II), as well as Zn(II). The metal-dependent ATP hydrolysis activity of purified ZntA was recently characterised and showed a specificity for Cd(II), Pb(II) and Zn(II). zntA expression has been reported to be up-regulated primarily by Zn(II), mediated by the regulatory protein ZntR, belonging to the MerR transcriptional regulator family. In contrast to previous claims, we now show, using a Phi(zntA-lacZ) monolysogen, that Cd(II) is the most effective inducer of zntA, which is also induced significantly by Pb(II). The Cd(II)- and Pb(II) dependent transcriptional up-regulation of zntA is also mediated by ZntR. PMID- 10802062 TI - Domain 2 of gelsolin binds directly to tropomyosin. AB - Gelsolin is an actin filament severing protein composed of six similar structured domains that differ with respect to actin, calcium and polyphospho-inositide binding. Previous work has established that gelsolin binds tropomyosin [Koepf, E.K. and Burtnick, L.D. (1992) FEBS Lett. 309, 56-58]. We have produced various specific gelsolin domains in Escherichia coli in order to establish which of the six domains binds tropomyosin. Gelsolin domains 1-3 (G1-3), G1-2 and G2 all bind tropomyosin in a pH and calcium insensitive manner whereas binding of G4-6 to tropomyosin was barely detectable under the conditions tested. We conclude that gelsolin binds tropomyosin via domain 2 (G2). PMID- 10802063 TI - Different membrane targeting of prostaglandin EP3 receptor isoforms dependent on their carboxy-terminal tail structures. AB - Mouse prostaglandin EP3 receptor consists of three isoforms, EP3alpha, beta and gamma, with different carboxy-terminal tails. To assess the role of their carboxy terminal tails in membrane targeting, we examined subcellular localization of myc tagged EP3 isoforms expressed in MDCK cells. Two isoforms, EP3alpha and EP3beta, were localized in the intracellular compartment but not in the plasma membrane, while the EP3gamma isoform was found in the lateral plasma membrane and in part in the intracellular compartment. Mutant EP3 receptor lacking the carboxy terminal tail was localized in the intracellular compartment but not in the plasma membrane. Thus, EP3 isoforms differ in subcellular targeting, and the carboxy-terminal tails play an important role in determination of the membrane targeting of EP3 receptor. PMID- 10802064 TI - Functional characterization of human sphingosine kinase-1. AB - Sphingosine kinase catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a novel lipid mediator with both intra- and extracellular functions. Based on sequence identity to murine sphingosine kinase (mSPHK1a), we cloned and characterized the first human sphingosine kinase (hSPHK1). The open reading frame of hSPHK1 encodes a 384 amino acid protein with 85% identity and 92% similarity to mSPHK1a at the amino acid level. Similar to mSPHK1a, when HEK293 cells were transfected with hSPHK1, there were marked increases in sphingosine kinase activity resulting in elevated SPP levels. hSPHK1 also specifically phosphorylated D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N, N dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol. Northern analysis revealed that hSPHK1 was widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. Thus, hSPHK1 belongs to a highly conserved unique lipid kinase family that regulates diverse biological functions. PMID- 10802065 TI - Caloric restriction prevents oxidative damage induced by the carcinogen clofibrate in mouse liver. AB - Long-term caloric restriction in rodents is known to decrease levels of oxidative damage, which may contribute to an 'anti-ageing' effect. We show here that a shorter period (10 months) of caloric restriction had only small effects on levels of oxidative DNA and protein damage in the livers of mice, but completely attenuated increased oxidative damage caused by the carcinogen clofibrate. Since clofibrate is thought to exert its actions by increasing oxidative damage, our data suggest that 10 months of caloric restriction can increase the resistance of tissues to agents inducing oxidative stress. This may be an important factor in explaining how caloric restriction decreases cancer incidence. PMID- 10802066 TI - Membrane anchoring and surface distribution of glycohydrolases of human erythrocyte membranes. AB - The membrane anchoring of the following glycohydrolases of human erythrocyte plasma membranes was investigated: alpha- and beta-D-glucosidase, alpha- and beta D-galactosidase, beta-D-glucuronidase, N-acetyl-beta-D-glucosaminidase, alpha-D mannosidase, and alpha-L-fucosidase. Optimized fluorimetric methods for the assay of these enzymes were set up. Treatment of the ghost preparation with 1.0 mol/l (optimal concentration) NaCl caused release ranging from 4.2% of alpha-D glucosidase to 70% of beta-D-galactosidase; treatment with 0.4% (optimal concentration) Triton X-100 liberated 5.1% of beta-D-galactosidase to 89% of alpha-D-glucosidase; treatment with 1.75% (optimal concentration) octylglucoside yielded solubilization from 6.3% of beta-D-galactosidase to 85% of alpha-D glucosidase. Treatment with phosphoinositide-specific phospholipase C caused no liberation of any of the studied glycohydrolases. These results are consistent with the notion that the above glycohydrolases are differently anchored or associated with the erythrocyte plasma membrane, and provide the methodological basis for inspecting the occurrence of these enzymes in different membrane microdomains. PMID- 10802067 TI - The Kruppel-like transcriptional factors Zf9 and GKLF coactivate the human keratin 4 promoter and physically interact. AB - Zf9/CPBP/KLF6 is a widely expressed member of the Kruppel-like family of transcriptional factors which regulates gene expression in hepatic stellate cells. Because of its ubiquitous expression including in the esophagus, we have explored its function in the esophageal squamous epithelium, a model system to study cellular proliferation and differentiation. Reverse transcription-PCR (RT PCR) and Western blot analyses revealed that Zf9 was highly expressed in human esophageal squamous cancer cell lines. Additionally, Zf9 localizes to the esophageal squamous epithelium by immunohistochemistry. Using transient transfection, Zf9 transactivates the human keratin 4 (K4) promoter reporter gene construct in a subset of the esophageal cancer cell lines through indirect mechanisms. Co-transfection of Zf9 and GKLF/KLF4, which is also a member of the Kruppel-like factors and expressed in the esophageal squamous epithelium, leads to coactivation in an additive fashion. Furthermore, we demonstrate that there is a physical interaction between GKLF and Zf9, a novel finding for Kruppel-like family members. PMID- 10802068 TI - G(i1) and G(oA) differentially determine kinetic efficacies of agonists for kappa opioid receptor. AB - We examined the diversity of single receptor function by measuring receptor-G protein coupling in the baculovirus-Sf21 expression system. In comparative studies using Sf21 cell membranes expressing kappa-opioid receptor (KOR) plus Galpha(i1)beta(1)gamma(2) or KOR plus Galpha(oA)beta(1)gamma(2), there was no significant difference between both preparations in the K(i) values of various kappa-opioid ligands for the displacement of [(3)H]U69593 binding. However, a marked difference in the rank order of agonists to stimulate [(35)S]GTPgammaS binding was observed between both preparations. These findings suggest that agonist efficacy is dependent on the population of different G proteins expressed in various tissues. PMID- 10802069 TI - Significant change in the structure of a ribozyme upon introduction of a phosphorothioate linkage at P9: NMR reveals a conformational fluctuation in the core region of a hammerhead ribozyme. AB - A modified hammerhead ribozyme (R32S) with a phosphorothioate linkage between G(8) and A(9), a site that is considered to play a crucial role in catalysis, was examined by high-resolution 1H and (31)P nuclear magnetic resonance (NMR) spectroscopy. Signals due to imino protons that corresponded to stems were observed, but the anticipated signals due to imino protons adjacent to the phosphorothioate linkage were not detected and the (31)P signal due to the phosphorothioate linkage was also absent irrespective of the presence or absence of the substrate. (31)P NMR is known to reflect backbone mobility, and thus the absence of signals indicated that the introduction of sulfur at P9 had increased the mobility of the backbone near the phosphorothioate linkage. The addition of metal ions did not regenerate the signals that had disappeared, a result that implied that the structure of the core region of the hammerhead ribozyme had fluctuated even in the presence of metal ions. Furthermore, kinetic analysis suggested that most of the R32S-substrate complexes generated in the absence of Mg(2+) ions were still in an inactive form and that Mg(2+) ions induced a further conformational change that converted such complexes to an activated state. Finally, according to available NMR studies, signals due to the imino protons of the central core region that includes the P9 metal binding site were broadened or not observed, suggesting that this catalytically important region might be intrinsically flexible. Our present analysis revealed a significant change in the structure of the ribozyme upon the introduction of the single phosphorothioate linkage at P9 that is in general considered to be a conservative modification. PMID- 10802070 TI - Wortmannin inhibits activation of nuclear transcription factors NF-kappaB and activated protein-1 induced by lipopolysaccharide and phorbol ester. AB - Whether all inflammatory agents activate nuclear transcription factors NF-kappaB and activated protein-1 (AP-1) through the same mechanism is not known. We examined the effect of the phosphatidylinositol-3-kinase (PI-3K) inhibitor wortmannin on the activation of NF-kappaB and AP-1 by different inflammatory agents. Wortmannin blocked NF-kappaB and AP-1 activation by lipopolysaccharide and phorbol ester but had minimal effect on activation by hydrogen peroxide, ceramide, okadaic acid and tumor necrosis factor. Inhibition of NF-kappaB correlated with abrogation of the degradation of IkappaBalpha and of NF-kappaB dependent reporter gene transcription. Thus, the mechanism of NF-kappaB and AP-1 activation by lipopolysaccharide and phorbol ester involves PI-3K. PMID- 10802072 TI - Corrigendum to: distinct mechanisms of antibody-mediated enzymatic reactivation in beta-galactosidase molecular sensors. PMID- 10802071 TI - The nitration of platelet cytosolic proteins during agonist-induced activation of platelets. AB - The nitration of protein tyrosine residues by peroxynitrous acid has been associated with pathological conditions. Here it is shown, using a sensitive competitive enzyme-linked immunosorbent assay and immunoblotting for nitrotyrosine, that spontaneous nitration of specific proteins occurs during a physiological process, the activation of platelets by collagen. One of the main proteins nitrated is vasodilator-stimulated phosphoprotein. Endogenous synthesis of nitric oxide and activity of cyclo-oxygenase were required for the nitration of tyrosine. The nitration was mimicked by addition of peroxynitrite to unstimulated platelets, although the level of nitrotyrosine formation was greater and its distribution among the proteins was less specific. PMID- 10802073 TI - Corrigendum to: the role of conserved extracellular cysteine residues in vasopressin V2 receptor function and properties of two naturally occurring mutant receptors with additional extracellular cysteine residues. PMID- 10802074 TI - Corrigendum to: A novel rod-like opsin isolated from the extra-retinal photoreceptors of teleost fish. PMID- 10802075 TI - Announcing the winner of the IJGO prize paper award for 1999. Estrogen replacement therapy in breast cancer survivors AB - Objective: To determine whether estrogen replacement therapy (ERT) adversely affected outcome of breast cancer survivors. Method: A prospective descriptive study of all breast cancer survivors who requested ERT because of intractable menopausal symptoms. All patients presented voluntarily as gynecological outpatients and were all given oral continuous opposed ERT: 20 premarin and medroxyprogesterone and four tibolone. Results: Twenty-four patients who had previously been treated for breast cancer 8-91 months prior to their initiating ERT have been observed for 24-44 months. There were 15 patients with stage 1, eight with stage 2 and one with stage 4 breast cancer. The mean age of the patients at commencement of ERT was 48 years (range 42-161). Two patients had a biopsy of a suspicious breast nodule: both of which were benign. There have not been any recurrences to date. Conclusion: Breast cancer survivors did not have their outcome adversely affected by ERT during an observation period of 24-44 months. PMID- 10802076 TI - Molecular prenatal diagnosis of thalassemia in Taiwan. AB - OBJECTIVE: Thalassemia is a highly prevalent genetic disorder in Taiwan. The major goal of this study was to present a feasible protocol for the prenatal diagnosis of thalassemia. METHOD: Prenatal investigation of thalassemia was performed on 57 at-risk cases at the Mackay Memorial Hospital, Taipei, Taiwan. We developed a method using polymerase chain reaction (PCR) and high-throughput DNA sequencing to detect mutations. All diagnoses were confirmed after delivery. RESULT: Prenatal testing revealed 16 normal fetuses, 24 alpha-thal-1 carriers, eight Hb Bart's hydrops fetalis, seven beta-thalassemia minor, and two beta thalassemia major fetuses. No false-positive or false-negative cases were found during the postnatal follow-ups. CONCLUSION: The results of this study indicate that prenatal diagnosis of thalassemia syndromes in Taiwan is successful with the use of a rapid and accurate molecular method. PMID- 10802078 TI - Transdermal glyceryl trinitrate in the management of primary dysmenorrhea. AB - OBJECTIVE: Increased intra-uterine pressure due to exaggerated myometrial contractions is an important factor in the pathogenesis of dysmenorrhea, its treatment being associated with uterine muscle relaxation. Diminished synthesis of endogenous nitric oxide has been shown to induce myometrial contractions and, conversely, the administration of exogenous nitric oxide has successfully resulted in uterine relaxation in a variety of obstetrical/gynecological disorders. The objective of this study was to determine the role of transdermal glyceryl trinitrate, as a source of exogenous nitric oxide, in the management of primary dysmenorrhea. METHOD: This was a multi-national, double-blind, randomized and cross-over study in patients with primary dysmenorrhea. Eighty-eight patients from six countries were evaluated during three menstrual cycles while receiving glyceryl trinitrate patches, 0.1 mg/h (x) or matching placebo patches. Pain intensity scores assessed on a visual analog scale and the time-weighted sum of the pain intensity differences (SPID) were evaluated during days 1, 2 and 3 of each cycle using an analysis of variance (ANOVA) model. Overall assessment of efficacy and the incidence of adverse events were analyzed by the Stuart-Maxwell or the McNemar tests as appropriate. RESULTS: Efficacy was determined for the first day of each cycle, all days/all cycles and for patients who completed at least one cycle in each treatment modality. In all three analyses, SPIDs were statistically superior (P<0.01) for the glyceryl trinitrate patches. Pain intensity differences from hours 1 to 6 also showed statistically significant differences in favor of the active treatment. In the overall assessment of efficacy, glyceryl trinitrate patches were statistically superior as well. The incidence of headache was 26% for the active drug and 6.1% for placebo (P<0.01). CONCLUSIONS: The data indicate that transdermal glyceryl trinitrate, as a source of exogenous nitric oxide, is useful as a modulator of uterine contractility representing, therefore, a new and mechanistically different therapeutic alternative for the management of primary dysmenorrhea. PMID- 10802077 TI - The routine use of cefazolin in cesarean section. AB - OBJECTIVE: To determine the effectiveness and safety of the routine use of antibiotic prophylaxis in women undergoing cesarean section. METHOD: Four hundred and forty-one women undergoing cesarean sections were randomly assigned either to a single dose of 1 g intravenous cefazolin or placebo after clamping of the umbilical cord. The primary outcome was the development of post-operative febrile morbidity and the secondary outcomes were infection-related complications. RESULT: There were 211 emergency and 230 elective cesarean sections. In the emergency cesarean sections, 34 (30.6%) women developed post-operative febrile morbidity in the placebo group compared to 11 (11%) women in the cefazolin group. This was a statistically significant difference (P = 0.001). Similarly, there were statistically significant differences between the two groups in the development of wound infection (P<0.001), use of therapeutic antibiotics (P = 0.001), and post-operative days in hospital (P = 0.003). No statistically significant differences were detected in the development of other infection related complications. In the elective cesarean sections, no statistically significant differences were found in post-operative febrile morbidity and infection-related complications. There were no serious side effects related to the use of cefazolin. CONCLUSION: The routine use of a single dose of cefazolin is safe and effective in emergency but not elective cesarean section. PMID- 10802079 TI - Endometrial prolactin in hyperprolactinemic women. AB - OBJECTIVE: To observe the behavior of endometrial prolactin in hyper- and normoprolactinemic women. METHOD: Forty women were selected due to alterations in menstrual cycle and/or galactorrhea and/or sterility. Patients were divided into two groups: Group A, 19 hyperprolactinemic women; and Group B, 21 normoprolactinemic women. Their mean age was 28.3 years. Laboratory assessments were carried out in the initial follicular phase (days 3-9), initial luteal phase (days 15-21), and late luteal phase (days 22-29) in menstruating women (samples 1, 2 and 3, respectively). In the non-menstruating women, samples were collected on days 1, 14 and 21 after the initial appointment (samples 1, 2 and 3, respectively). LH, FSH, prolactin and estradiol were measured in sample 1, while prolactin and progesterone were measured in samples 2 and 3. All women were submitted to two biopsies of the endometrium for observing the menstrual cycle, dating, and immunohistochemical detection of endometrial prolactin. Biopsies were obtained simultaneously to samples 2 and 3. To compare the two groups, Student's t-tests or Mann-Whitney statistics were performed. RESULT: Results showed that the mean percentage of endometrial prolactin were higher in the late luteal phase compared to the initial one in hyper- and normoprolactinemic women. CONCLUSION: The data allowed us to conclude that serum prolactin is not correlated to endometrial prolactin, and that the synthesis of the latter is directly correlated to the differentiation of stromal cells, that is induced by progesterone in a decidualized endometrium. PMID- 10802080 TI - Basal leptin concentrations in women with normal and dysfunctional ovarian conditions. AB - OBJECTIVE: To determine whether leptin is involved in ovarian function. METHODS: Fasting serum samples were obtained from 20 women with normal menstrual cycles who were either obese or non-obese: 12 non-obese patients with polycystic ovary syndrome (PCOS), 8 obese patients with PCOS, 10 patients with stress-related hypothalamic amenorrhea, and 8 patients with weight loss-related hypothalamic amenorrhea. RESULTS: Serum leptin levels were strongly related to body mass index (BMI) in each group, but there was no difference in the mean serum leptin levels among the BMI-matched study groups. A significant difference in the mean serum leptin levels was found between the non-obese and obese control groups (P<0.001) and between the non-obese and obese PCOS groups (P<0.001). CONCLUSIONS: These findings indicate that circulating leptin levels in women with normal menstrual cycles and those with ovarian dysfunction are strongly related to BMI. Leptin does not appear to be primarily involved in regulating ovarian function. PMID- 10802081 TI - Simplification of the clinical phase of IVF and ICSI treatment in programmed cycles. AB - OBJECTIVE: To evaluate the success of a protocol for controlled ovarian hyperstimulation allowing patient self-selection into groups for ovulation stimulation planned 8 weeks and more in advance following cycle synchronization, drug self-administration as well as a reduced number of folliculometries. METHODS: A total of 714 patients received the same stimulation protocol. In 260 cases GnRH-a was applied daily and in 454 as depot. In all patients FSH-HP was self-administered subcutaneously for ovarian stimulation. In 316 patients IVF and in 398 patients ICSI was performed. RESULTS: The delivery rate per started cycle was higher in patients receiving depot GnRH-a in the IVF and ICSI group (30.2 vs. 23.4) than in those receiving subcutaneous GnRH-a (20.2 vs. 22.1). CONCLUSION: Programming of the IVF/ICSI cycle greatly simplifies treatment. A comparison of pregnancy rate and delivery rate per cycle between depot and subcutaneous daily application of GnRh-a did not confirm any statistically significant difference. PMID- 10802082 TI - Cost-effectiveness of IVF in women 38 years and older. AB - OBJECTIVE: To compare the cost per delivery in women younger than 38 years with women equal to or older than 38 years of age attempting IVF. METHODS: All couples undergoing IVF treatment between October 1991 and September 1998 were enrolled in this study. A standard protocol of controlled ovarian hyperstimulation was employed throughout the study. Four hundred and seven cases were allocated to two groups - group I composed of patients younger than 38 years of age and group II of patient equal to or older than 38 years of age. The total cost of each successful outcome was the goal of our study. RESULTS: A total of 407 women underwent 722 stimulated cycles for IVF of which 122 cycles (16.89%) did not proceed to oocyte retrieval. We found statistically significant differences in the cancellation rate, the number of hMG ampoules, the number of oocytes retrieved, the number of oocytes fertilized, the number of embryos transferred, the clinical pregnancy rate, the rate of multiple pregnancy, the delivery per initiated cycle and the cost per delivery between the two groups (P<0.05, significant). The cost per delivery in group II was approximately 3.6 times that of group I. CONCLUSIONS: Women age 38 years or more have less chance of a successful outcome from IVF treatment. Couples contemplating IVF should be provided with accurate information about prognosis for the pregnancy and the financial costs. PMID- 10802083 TI - Partial hydatidiform mole with diploid karyotype in a live fetus. AB - Even though most instances of partial mole are triploid, only a few cases of diploid partial moles have been reported. Prognosis of partial mole is usually better than the complete mole as few cases of partial moles progress to persistent trophoblastic disease. However, the nature and the risks of diploid partial moles are not well established and they seem to be a distinct clinical entity. Here we report a case of partial mole presenting with a 22 weeks live fetus and preeclampsia. Fetal blood sampling was performed for cytogenetic analysis which revealed a diploid (46XY) karyotype. No progression to malignant gestational trophoblastic disease occurred. PMID- 10802084 TI - Impetigo herpetiformis at the 36th week of gestation. PMID- 10802085 TI - Shared antenatal care to increase utilization of maternity services. PMID- 10802086 TI - Unexplained stillbirths: are they preventable? PMID- 10802087 TI - Three-dimensional sonographic features of fetal sacrococcygeal teratoma. PMID- 10802088 TI - Hysteroscopic metroplasty: section of the cervical septum does not impair reproductive outcome. PMID- 10802089 TI - Secondary infertility and dysfunctional uterine bleeding from a Lippes loop placed 32 years earlier. PMID- 10802090 TI - Adult respiratory distress syndrome as a manifestation of ovarian hyperstimulation syndrome. PMID- 10802091 TI - Menstruation-induced uterine rupture. PMID- 10802092 TI - Nuchal cord encirclements and risk of stillbirth. PMID- 10802093 TI - Prevalence of HPV infection in cervical cytology-normal women in Okinawa, Japan, as determined by a polymerase chain reaction. PMID- 10802094 TI - Non-oral HRT and lipoprotein metabolism. Which progestin-- dydrogesterone or medroxyprogesterone acetate? PMID- 10802095 TI - Dehydration in dormant insects. AB - Many of the mechanisms used by active insects to maintain water balance are not available to dormant individuals. Physiological and biochemical mechanisms of dehydration tolerance and resistance in dormant insects and some other invertebrates are reviewed, as well as linkages of dehydration with energy use and metabolism, with cold hardiness, and with diapause. Many dormant insects combine several striking adaptations to maintain water balance that-in addition to habitat choice-may include especially reduction of body water content, decreased cuticular permeability, absorption of water vapour, and tolerance of low body water levels. Many such features require energy and hence that metabolism, albeit much reduced, continues during dormancy. Four types of progressively dehydrated states are recognized: water is managed internally by solute or ion transport; relatively high concentrations of solutes modify the behaviour of water in solutions; still higher concentrations of certain carbohydrates lead to plasticized rubbers or glasses with very slow molecular kinetics; and anhydrobiosis eliminates metabolism. PMID- 10802096 TI - Identification of AB - The neuropeptides inducing dark color in albino nymphs of the migratory locust Locusta migratoria were isolated from the larval brain of the silkworm, Bombyx mori and from the adult corpora cardiaca (CC) of the cricket Gryllus bimaculatus, respectively, and their amino acid sequences identified. The two peptides isolated from the two different species are identical to [Arg(7)] corazonin, a neuropeptide known to be present in a cockroach and others. This peptide induces a dark color in albino nymphs of L. migratoria at fmol levels, and a high dose of >/=100 pmol caused albino locusts to turn completely black, but it influenced neither body color nor metamorphosis in B. mori and G. bimaculatus. Therefore, the physiological functions of [Arg(7)] corazonin in the silkworm and the cricket remain unknown. The present study demonstrated the usefulness of the albino strain of L. mirgatoria as a specific bioassay system for this peptide. PMID- 10802097 TI - The influence of parasitism on wing development in male and female pea aphids. AB - Wing formation in presumptive alate morphs (virginoparae and males) was observed for the pea aphid, Acyrthosiphon pisum, exposed to attack by the parasitoid, Aphidius ervi, at different stages of host development. Morphological abnormalities in parasitized aphids such as complete apterization (development of a wingless form), formation of rudimentary wing buds, and deformed wings indicate a possible disruption of the endocrine system. Changes in the body shape and the number of olfactory secondary rhinaria on the antennae could indicate an influence of juvenile hormone in parasitized A. pisum but the development of fifth-stadium supernumerary larvae (indicated by an extra moult and which can be induced by exogenous juvenile hormone treatments) was not found in parasitized aphids. In addition, while apterization of virginoparae can also be induced by the pro-allatocidal compound Precocene III, this was not possible in the male. Males which survived parasitoid attack without forming aphid mummies (indicating that oviposition had not occurred) developed as wingless individuals suggesting that the reproductive-tract-fluids from the female parasitoid were important in the wing inhibition process. Teratocytes from the parasitoid appeared to promote developmental arrest in parasitized aphids. PMID- 10802098 TI - Water relations of the freeze-tolerant New Zealand alpine cockroach Celatoblatta quinquemaculata (Dictyoptera: Blattidae). AB - Celatoblatta quinquemaculata is a freeze-tolerant alpine cockroach found on the Rock and Pillar Range, Central Otago, New Zealand. This study investigated seasonal changes in water content, as well as desiccation tolerance, and the relationship between desiccation and cold tolerance. Whole body water contents from field-fresh cockroaches collected over a 20 month period ranged from 69.9+/ 1.0% fresh weight (FW) in February 1998 to 60.3+/-1.1% FW in July 1998. Water contents were significantly lower in winter than summer, and were positively correlated to microhabitat temperatures over the week preceding collection. Cockroaches survived the loss of up to 82% (mean: 56.7%+/-10.2) of their initial body water content, and the amount of water loss sustained was not dependent on the rate of water loss. Cockroaches did not suffer further mortality due to desiccation after removal to 99% relative humidity, but only regained lost water if given access to liquid water. Experimental dehydration did not enhance freeze tolerance, but did slightly lower the supercooling point. It is concluded that reduction of body water content in winter may be a consequence of cold hardening responses, but desiccation does not constitute the cold hardening mechanism itself. PMID- 10802099 TI - Odour representation in honeybee olfactory glomeruli shows slow temporal dynamics: an optical recording study using a voltage-sensitive dye. AB - Stimulation with odours has been shown to elicit characteristic patterns of activated glomeruli in the antennal lobe (AL) of honeybees. In this study we show that these patterns are dynamic in a time window of 2-3 s after stimulus onset. We measured changes in the averaged membrane potential of all cells in the glomerular neuropil by optical imaging of the voltage-sensitive dye RH795 using a slow scan CCD camera (3 frames/s). The four substances 1-hexanol, hexanal, citral and clove-oil as well as the binary mixtures hexanol+hexanal and hexanol+citral were used as stimuli (2 s stimulus duration). We found that: (1) every odour elicited an odour-specific activity pattern, and conversely every glomerulus had a characteristic odour response profile; (2) some glomeruli had a tonic, some a phasic-tonic, and some a slow phasic response pattern; (3) the difference between the glomerular response patterns increased within 2 s of stimulus presentation, which suggests that odour representations became more characteristic over stimulus time; and (4) the responses to odorant mixtures were complex and glomerulus-dependent: some responses correspond to the sum of the compounds' responses, some to the response of one of the components. PMID- 10802100 TI - Circadian component influences the photoperiodic induction of diapause in a drosophilid fly, Chymomyza costata. AB - The last-instar larvae of a drosophilid fly, Chymomyza costata enter diapause in response to the dark-phases longer than 9 h (Yoshida, T., Kimura, M.T., 1995. The photoperiodic clock in Chymomyza costata. Journal of Insect Physiology 41, 217 222). In order to switch the developmental programming of the sensitive larvae from continuous development to diapause, after they were transferred from the short (8 h) to the long (14 h) dark-phase, significantly less time (1-2 days) was required when the dark-phase was abruptly and asymmetrically extended into the evening, than when it was extended symmetrically into both morning and evening (2 3 days), or asymmetrically into the morning hours (4-6 days). Diapause was also induced in 40-70% of sensitive larvae that were reared under the gradually shortening light-phase (from 16 h to 2 h, by 1 h in each cycle), despite that the dark-phase remained constant and short (8 h). Larvae developed continuously, however, when reared under the gradually extending light-phase (from 16 h to 24 h) and a constantly short dark-phase. We interpret such results, with the help of the two-oscillator model of circadian rhythmicity (Pittendrigh, C.S., Daan, S., 1976. A functional analysis of circadian pacemakers in nocturnal rodents. V. Pacemaker structure: A clock for all seasons. Journal of Comparative Physiology A 106, 333-355), as indicating that two mutually coupled oscillators (evening and morning) differing in their entrainability may participate in measuring of the dark-phase duration. The levels of dopamine (DA) and serotonin (5-HT) in the larval CNS transiently increased (by up to 20%) after the dusk, while no apparent change was observed during the dawn. The dusk-related increase was observed also after the asymmetric extension of the dark-phase into evening, while the asymmetric extension into morning had no effect on the levels of the DA and 5-HT. PMID- 10802101 TI - Oxygen levels in the gut lumens of herbivorous insects. AB - Oxygen levels were measured in the foregut and midgut lumens of ten species of caterpillars and three species of grasshoppers. In most species, the foregut was nearly anoxic, with oxygen levels ranging from 0 to 2.5 mm Hg. However, two caterpillar species with large foreguts (Malacosoma disstria and Lymantria dispar) had elevated oxygen levels (27.9 and 32.1 mm Hg) in this region when they were fed artificial diet. In all of the species surveyed, the anterior and posterior midgut were nearly anoxic, with oxygen levels ranging from 0 to 7.3 mm Hg. Oxygen levels in the midgut lumen of Helicoverpa zea did not differ when caterpillars were fed artificial diet or tomato foliage, suggesting that the insect is capable of reducing the level of ingested oxygen in its gut. An examination of the radial microgradient of oxygen in the gut lumen demonstrated that the midgut epithelium is not a sink for ingested oxygen. However, the midgut contents of larvae fed artificial diet were capable of depleting oxygen. This capacity was reduced by boiling, suggesting that the nearly anoxic state of the midgut lumen in some insects is maintained by endogenous chemical processes. We conclude that low oxygen levels in the gut lumens of most herbivorous insects may greatly reduce the rates of oxidation of ingested plant compounds by oxygen dependent reactions. PMID- 10802102 TI - A neurophysiological study of sensitivity to a feeding deterrent in two sister species of Heliothis with different diet breadths. AB - Heliothis subflexa and H. virescens are sister species that differ markedly in their hostplant specificity: the former is a specialist on one plant genus, the latter feeds on plants from many families. The behavioral threshold for rejection of deterrent chemicals is lower in larvae of H. subflexa than in those of H. virescens. In this paper, we examine the responses of the galeal styloconic sensilla of these larvae to stimulation by three chemicals, sucrose and inositol, which are phagostimulants, and sinigrin, a deterrent, in an attempt to determine the neural basis for the differences in feeding behavior between the species. The species difference could not be attributed to differences in firing rate of the deterrent-sensitive cells, differences in the ratio of responses to phagostimulants and deterrents, differences in the rates of adaptation of the sensory neurons, or differences in the extent of interactions between chemicals at the peripheral sensilla. We conclude that the differences between the species probably result from differences in processing sensory information within the central nervous system. PMID- 10802103 TI - Regulation of pheromone inhibition in mated females of Choristoneura fumiferana and C. rosaceana. AB - In the spruce budworm, Choristoneura fumiferana, and the obliquebanded leafroller, C. rosaceana, mating significantly depressed pheromone production after 24 h. On subsequent days, the pheromone titre increased slightly in C. fumiferana, but not in C. rosaceana. No pheromonostatic activity was associated with male accessory sex gland (ASG) extracts, 20-hydroxy-ecdysone or hemolymph taken from mated females. However, pheromone production in mated females was not suppressed when the ventral nerve cord (VNC) was transected prior to mating, indicating that an intact VNC is required to permanently switch off pheromone production after mating. As suggested for other moth species, the presence of sperm in the spermatheca probably triggers the release of a signal, via the VNC, to inhibit pheromone production. The fact that in both species the brain suboesophageal ganglion (Br-SEG) of mated females contains pheromonotropic activity and that their pheromone glands may be stimulated by the synthetic pheromone-biosynthesis-activating-neuropeptide (PBAN) or a brain extract supports the hypothesis that the neural signal prevents the release of PBAN into the hemolymph rather than inhibiting its biosynthesis. Therefore, we speculate that following the depletion of sperm in the spermatheca, the neural signal declines and is less effective in preventing the release of PBAN, thereby stimulating the resumption of pheromone production, as seen in mated C. fumiferana females. In a previous study, mating was shown to induce a significant rise in the juvenile hormone (JH) titre of both Choristoneura female moths, suggesting that post mating pheromone inhibition may be under hormonal regulation. However, following topical applications or injections of the juvenile hormone analogue (JHA) and JH II into virgins, the pheromone only declined significantly 48 h after treatment in C. rosaceana. This suggests that the significant rise in the hemolymph JH titre after mating in C. rosaceana females plays a role in keeping the pheromone titre consistently low throughout their reproductive life. These findings will be discussed in relation to the different life histories of the two Choristoneura species. PMID- 10802104 TI - Effects of chilling stress on allatal growth and juvenile hormone synthesis in the cockroach, Diploptera punctata. AB - During the ovarian cycle of the cockroach, Diploptera punctata, a mitotic wave occurs in the corpora allata before an increase in gland volume and juvenile hormone (JH) synthesis. Previous studies have demonstrated that the brain inhibits mitosis and JH synthesis in corpus allatum (CA) cells until adult females have mated. Herein, we report that chilling stress effectively suppresses mating induced proliferation of CA cells. In mated females, chilling on melting ice for 0.5-3 hours caused a strong, dose-dependent decrease in mitotic activity. In insects chilled for 3 hours, although the mitotic wave in the CA was practically abolished, CA volume and JH synthesis finally reached peak levels typical of unchilled insects, despite a 2-day delay. Consequently, oocyte maturation and oviposition were also delayed by 2 days, yet in both chilled and unchilled insects, peak values of basal oocyte length were the same. By allowing virgin females to mate on different days after chilling, we found that the chilling effect could be retained in the insect body for at least 2 days. During this period, signals from mating could not effectively remove inhibition of CA cell proliferation. Unilaterally disconnecting the CA from the brain revealed that chilling stress mediated CA cell proliferation via the brain, and did not directly affect the CA. PMID- 10802105 TI - Behavioral and developmental homeostasis in the fire ant, Solenopsis invicta. AB - The von Bertalanffy rule (1960) predicts that low incubation temperature during larval development will result in larger adult body size. If larval development in social insects followed this rule, then low incubation temperature would induce the development of larger workers and possibly even sexuals. To test this prediction, the effect of incubation temperature on larval development, larval meal size, larval tending and worker recruitment to food in the fire ant, Solenopsis invicta was investigated. Temperatures tested where within the range at which brood remains viable.Contrary to the predictions of the von Bertalanffy rule, worker size was unaffected by incubation temperature, and sexuals were reared at the high rather than the low incubation temperature. Moreover, larval meal size, the rate of larval tending by workers and the total number of workers recruited to food were unaffected by temperature.Mechanisms regulating developmental and behavioral homeostasis were as follows: the duration of larval development and the rate of larval growth changed proportionately with temperature such that the mean and variation of pupal size was unaffected by incubation temperature. Larvae solicited at the same rate, swallowed at the same rate and swallowed for the same duration such that meal size was unaffected by incubation temperature. On the brood pile, fewer workers tended brood at higher incubation temperatures, but worker tempo increased; as a result, brood tending was not adversely affected by incubation temperature. The rate of worker recruitment to food sites outside the nest increased with temperature, but the duration of the recruitment effort decreased such that, over time, the same total number of workers was employed to retrieve food.Incubation humidity was also investigated. When brood chambers were less than 100% humid, workers recruited to food and tended larvae (retrieved, sorted and groomed them), but did not feed larvae. Eventually, larvae died of starvation and were cannibalized. PMID- 10802106 TI - Role of feeding in the reproductive 'group effect' in females of the German cockroach Blattella germanica (L.). AB - We have found that whether a female German cockroach, Blattella germanica (L.), is kept alone or in the presence of another female has a major impact on how fast it reproduces and how much it eats. By the sixth day of adulthood, females paired since adult eclosion had substantially larger oocytes than did females isolated during the same time, and females paired with intact females, or with ones rendered incapable of feeding, consumed more rat chow in the first six days of adulthood than did isolated females. The stimulatory effect of pairing on reproduction was, however, partially independent of feeding because the oocytes of solitary and paired females differed in size on day 6 even when they were given, and had consumed, the same amount of food. This result was confirmed with analysis of covariance using the total food intake of a female as the covariate in the analysis. A female's social condition probably influenced the development of its oocytes by affecting the quantity of juvenile hormone synthesized by its corpora allata. The corpora allata of paired females produced more hormone than did those of isolated ones, even when all females had consumed an equivalent amount of food. Moreover, females treated with a juvenile hormone analog, fenoxycarb, reproduced more quickly than identically reared and fed control females, showing that juvenile hormone could influence reproduction independently of feeding. We conclude that both group rearing and food intake accelerate oocyte development by diminishing the brain's inhibition on the synthesis of juvenile hormone. PMID- 10802107 TI - Hexameric storage proteins during metamorphosis and egg production in the diamondback moth, Plutella xylostella (Lepidoptera). AB - As in many Lepidoptera, Plutella xylostella adults do not feed on protein and females must use accumulated reserves to supply vitellogenin synthesis. Storage proteins were quantified in females and males from the late larval stage through day 4 of adult life. The level of storage protein peaked in the early pupal stage, with females having about twice as much as males. In males, the level fell through pupal development and dropped to a trace by one day after eclosion. In females, level of storage proteins fell until eclosion, and then rose dramatically within four hours after the molt to about 2/3 of the original peak level. This post-eclosion increase, which has not been reported previously in insects, suggests that adult females synthesize hexamerins to resequester amino acids. Subsequently, the level of storage proteins fell as vitellogenin appeared and eggs were laid. The ability to synthesize and sequester amino acids as storage proteins during the adult stage has wide-ranging implication for protein management in insects, particularly those that are long-lived and have flexible schedules of reproduction. PMID- 10802108 TI - Evidence that dilation of isolated salivary ducts from the tick Dermacentor variabilis (Say) is mediated by nitric oxide. AB - We used pharmacological methods to test the hypothesis that female Dermacentor variabilis salivary ducts dilated when dopamine-stimulated and that dilation was nitric oxide-mediated. Stimulation with dopamine resulted in an increased diameter (19.7%) compared to unstimulated ducts (P<0.005). Pretreatment with L NAME, an inhibitor of nitric oxide synthase, or cytochalasin D abolished the dilation. Addition of L-arginine to L-NAME-treated ducts partially restored the ability to dilate. A cuticular coil composed of a series of concentric rings ran the length of the duct adjacent to the epithelial cell layer. In stimulated ducts, the center-to-center periodicity of these rings increased from 0.59 um in unfed ducts to 1.0 um from partially fed ducts (P<0.05). When the ducts from partially fed females were stimulated with dopamine, the periodicity increased further to 1.75 um (P<0.05), suggesting the coils moved further apart in response to stimulation. Prominent folds lining the lumen of unstimulated ducts were less pronounced in stimulated preparations, suggesting that the cuticle stretches, thereby increasing lumen size. Actin was localized in epithelial cells as a honeycomb pattern that we suggest links the epithelial cells to the rings. Together, these data support the following hypothesis: stimulated ducts dilated during fluid production; dilation involved an actin-based system, and was mediated by nitric oxide. Dilation of the duct may enhance its role as a reservoir for saliva produced by the acini during the period between imbibition and salivation. PMID- 10802109 TI - Action spectra of the female's response in the firefly Photinus pyralis (Coleoptera: Lampyridae): evidence for an achromatic detection of the bioluminescent optical signal. AB - Behavioural action spectra of the threshold of the Photinus pyralis female response to light stimuli simulating the bioluminescent optical signal of the conspecific male firefly were determined in the laboratory. The action spectra (Fig. 1) were narrow and peaked in the yellow region of the spectrum. The females responded only to stimuli of wavelengths longer than 480 nm and not to stimuli in the blue (420-460 nm) part of the spectrum. The shape of the function corresponds with (a) the electroretinographic spectral sensitivity function in the long wavelength (520-660 nm) region of the spectrum, (b) the action spectrum of the female response (Fig. 1), (c) the species yellow bioluminescence emission spectrum and (d) the action spectrum of the intracellular response from single retinular cells (Fig. 2) of the compound eyes in the firefly. Such a correspondence suggests that the narrow yellow receptors of the female mediate the detection and processing of the optical signal of the conspecific male. Since the bioluminescent optical signal is processed exclusively by a single receptor class, signal detection is achromatic. PMID- 10802110 TI - Regulation of vitellogenesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae). AB - Studies were undertaken to investigate vitellogenesis and its regulation in female adults of the fall armyworm, Spodoptera frugiperda. A single female specific protein, likely to be the S. frugiperda vitellogenin (Vg), appeared approximately 5 h after adult eclosion in the hemolymph of virgin females. The concentration of the protein increased with age as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed. A protein with the same relative molecular mass was also present in egg extracts, but absent from hemolymph samples from male moths. The relative molecular mass of the designated S. frugiperda Vg was determined as 164.5+/-2.5 kDa. Vitellogenic oocytes became visible 36-48 h after emergence and egg deposition began on day 3 of adult life. Vg could not be detected in the hemolymph of females decapitated directly after eclosion. When decapitated virgin females were injected with the JH-mimic methoprene (MP), the level of Vg was comparable to that in non-decapitated moths, indicating that vitellogenesis in S. frugiperda depends on juvenile hormone (JH). However, the number of vitellogenic oocytes was somewhat lower than in non decapitated virgin females. Injection of 20-hydroxyecdysone (20E) promoted Vg production to a similar extent in decapitated female moths, but in contrast to methoprene injection, treatment with 20E never resulted in the production of vitellogenic oocytes. In vitro cultivated ovaries of adult females dissected directly after eclosion produced lower amounts of ecdysteroids than those isolated on day 1 after emergence. Our results suggest a crucial role for 20E in the induction of vitellogenesis in the noctuid S. frugiperda, while JH seems to be essential for the continued uptake of Vg by developing oocytes and may trigger 20E biosynthesis in the ovary. PMID- 10802111 TI - Extracardiac versus cardiac haemocoelic pulsations in pupae of the mealworm (Tenebrio molitor L.). AB - Pulsations in mechanical pressure of the pupal haemocoele were investigated by means of simultaneous recording from multiple sensors. It has been determined that cardiac and extracardiac haemocoelic pulsations are each regulated by substantially different and quite independent physiological mechanisms. At the beginning and in the middle of the pupal interecdysial period the anterograde heartbeat and extracardiac pulsations occur in similar, but not identical periods. During the advanced pharate adult stage, there appear almost uninterrupted pulsations from different sources: cardiac, extracardiac, intestinal, and the ventral diaphragm.Extracardiac pulsations are associated with pressure peaks of 200-500 Pa, occurring at frequencies of 0.3-0.5 Hz. The effect of heartbeat on haemocoelic pressure is very small, 100- to 500-fold smaller, comprising only some 1 or 2 Pa during the vigorous anterograde systolic contractions. Accordingly, extracardiac pulsations are associated with relatively large abdominal movements from 30-90 um whereas heartbeat produces movements of only 100-500 nm. This shows that extracardiac pulsations can be easily confused with the anterograde heartbeat. It does not seem realistic to assume that the relatively weak insect heart, and not the 100- to 500-fold more powerful extracardiac system of abdominal pump, could be at all responsible for selective accumulation of haemolymph in anterior parts of the body, for inflation of wings or enhancement of tracheal ventilation.It has been established that thermography from the pericardial region is not specific for the heartbeat. It records subepidermal movement of haemolymph resulting from the actions of both dorsal vessel and extracardiac pressure pulses as well. Shortly before adult eclosion the cardiac and extracardiac pulsations occasionally strike in concert, which profoundly increases the flow of haemolymph through pericardial and perineural sinuses. The relatively strong extracardiac pulsations cause passive movements of various visceral organs, tissue membranes, or tissue folds, giving thus a false impression of an authentic pulsation of tissues. In addition, extracardiac pulsations cause rhythmical movements of haemolymph between various organs, thus preventing haemolymph occlusion at the sites where the heart does not reach. It has been emphasized, finally, that the function of the autonomic nervous system (coelopulse), which integrates extracardiac pulsations, depends on homeostatic moderation of excessive or deficient conditions in insect respiration and haemolymph circulation. PMID- 10802112 TI - Plasticity in caste-related exocrine secretion biosynthesis in the honey bee (Apis mellifera). AB - Plasticity of Dufour's gland secretion in the honey bee is correlated with the individual's plasticity. Queens and queenless (QL) egg-laying workers possess a bouquet of esters and hydrocarbons, whereas queenright (QR) workers produce exclusively hydrocarbons. The effects of social environment (QR vs. QL conditions) and possible physiological constraints on the gland were studied by following the biosynthesis of these classes of compounds in vivo and in vitro. Biosynthesis in vivo followed the prediction based on glandular chemistry. Queens and QL egg-laying workers, but not QR workers or QL foragers, showed incorporation of sodium acetate into both hydrocarbons and esters. In contrast, the in vitro studies revealed that, in addition to queens and QL egg-laying workers, QR nurses retained their ability to produce the queen characteristic esters. Although there was some ester production in foragers, it occurred to a lesser extent. It is possible that the glands in the older foragers undergo irreversible changes. The in vitro incubation also revealed a temporal activation of ester biosynthesis in QR workers. In these glands alcohols, corresponding to the alcohol moiety of the esters, predominated in short-term incubations but decreased as the amount of newly synthesized esters increased. In contrast, queens and QL egg-laying workers showed predominant incorporation into esters from the onset of incubation. Thus, expression within the workers' Dufour's gland is regulated. In the presence of a queen, ester production is inhibited. Once the queen is removed the physiologically unconstrained gland starts to biosynthesize the queen-specific esters after a certain lag needed for the build-up of precursors and the enzymatic machinery. PMID- 10802113 TI - Midgut-based resistance of Heliothis virescens to baculovirus infection mediated by phytochemicals in cotton. AB - The decrease in susceptibility to polyhedrosis disease when Heliothis virescens larvae feed on cotton is profound, limiting the utility of baculoviruses for controlling noctuids on this important crop. We observed that the mortalities of H. virescens larvae challenged with a reporter-gene construct of Autographa californica M nucleopolyhedrovirus (AcMNPV-hsp70/lacZ) and fed either lettuce or artificial diet were approximately 2.5-fold higher than that of cotton-fed insects. This decrease in susceptibility on cotton was observed following oral but not intrahemocoelic inoculation of virus, and it was negatively correlated with levels of foliar peroxidase. The rates of development of both infected and uninfected larvae also were correlated negatively with levels of foliar peroxidase, and hence, were significantly lower for insects fed cotton. When Calcofluor White M2R, an optical brightener reported to enhance the retention of AcMNPV-infected midgut cells, was included in inoculum administered orally to larvae, mortality levels were equivalent regardless of diet. These results suggest that sloughing of infected midgut cells occurred at a higher rate in insects that fed on cotton compared to the other two diets, and that midgut cell sloughing is the mechanism whereby susceptibility to mortal infection by AcMNPV hsp70/lacZ is decreased on cotton. This conclusion is consistent with previous reports that ingestion of cotton can generate reactive oxygen species within the midgut lumen that may damage midgut epithelial cells. As far as we know, this is the first study to link resistance intrinsic to the physiology of the insect (e.g., developmental resistance) and resistance conferred by host plant chemistry to a single mechanism, i.e., midgut cell sloughing. PMID- 10802114 TI - Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori. AB - Recently, it was confirmed that embryos derived from diapausing eggs of the silkworm, Bombyx mori, begin their development and reach larval maturity on mulberry leaves, when the naked eggs are cultured in vitro. In this study, we found that the method of embryo culture is useful for determining the physiological regulation of diapause. We show that the development of embryos derived from diapausing eggs was strongly inhibited by the addition of either sorbitol or trehalose to the culture medium. Furthermore, this inhibitory effect disappeared when the embryos were cultured in a control medium which did not contain either sorbitol or trehalose, indicating that the inhibitory reactions caused by both substances are reversible. The minimal effective dose of either sorbitol or trehalose was approximately 0.2 M, a value similar to the in vivo concentration of sorbitol in diapausing eggs (0.2 M). Glycerol, mannitol or glucose were moderately effective for inhibition. Sorbitol present in diapausing silkworm eggs does not appear to serve as an antifreeze, but as an strong arresting factor of embryonic development. Furthermore, these results show that a decrease in sorbitol releases the embryos from diapause at the termination of diapause. PMID- 10802115 TI - The role of cercal sensory feedback during spermatophore transfer in the cricket, Acheta domesticus. AB - The role of the cerci in the spermatophore transfer behavior of the cricket Acheta domesticus was examined. During transfer, the male cerci were held close to the female abdomen where they produced small flicking movements. Male cercal ablation significantly decreased mating success by reducing both the ability of the male to hook the epiphallus on to the female subgenital plate and to transfer the spermatophore. During spermatophore transfer, the male must thread the spermatophore tube into the female genital papilla and attach the spermatophore, via its attachment plate, to the base of the ovipositor. Extracellular recordings from the male genital nerve revealed that a centrally driven, rhythmic bursting activity of genital efferents produced the rhythmic contractions of the five pairs of genital muscles responsible for spermatophore threading. Tactile stimulation of campaniform sensilla on the medial aspect of each cercus inhibited the activity of those motor units responsible for advancing the spermatophore tube during threading, while simultaneously activating the motor units responsible for adjusting the position of the epiphallus. We conclude that mechanosensory neurons on the cerci of the male cricket supply important information on female position to the motor program responsible for spermatophore threading and transfer. PMID- 10802116 TI - Ecdysteroid titer, ovary status, and dominance in adult worker and queen bumble bees (Bombus terrestris). AB - The role of ecdysteroids in the regulation of dominance and reproduction in social Hymenoptera is little explored. In the current study we compared ecdysteroid titers in hemolymph of individual queen and worker bumble bees (Bombus terrestris) that differ in their behavior, reproductive status and social environment. Egg-laying queens that head colonies and have ovaries exhibiting all stages of follicle development, had a higher ecdysteroid titer than virgin queens whose ovaries contained only follicles at initial stages. In workers, the relationship between ecdysteroid titers and follicle development appears to be more complex and to be influenced by the bee's social status and social environment. Shortly after emergence, young workers had only follicles at the initial stages of oogenesis and they exhibited a low ecdysteroid titer. No significant correlation was detected between ovary status and ecdysteroid titer in workers, with some workers showing activated ovaries but low ecdysteroid titers. However, at six days of age, a trend towards higher ecdysteroid titer was observed for workers in queenless groups, a condition characterized by rapid follicle development relative to queenright conditions. In these queenless groups, high social status was associated with high ecdysteroid titers. By contrast, in queenright workers ecdysteroid titers were low, even for bees with presumably high social status that had activated ovaries and were observed performing oviposition behavior. This study suggests that ecdysteroids are involved in regulation of reproduction in B. terrestris. PMID- 10802117 TI - Host castration by Aphidius ervi venom proteins. AB - The braconid Aphidius ervi Haliday (Hymenoptera, Braconidae) is an endophagous parasitoid of the pea aphid, Acyrthosiphon pisum (Harris) (Homoptera, Aphididae). Parasitized host aphids show different degrees of castration, a response that is total when parasitoid oviposition takes place in first instar hosts. Deleterious effects on the host reproductive system are already evident by 24h following parasitization, before egg hatching. The effect of parasitoid venom on A. pisum ovaries has been studied by performing microinjections in non-parasitized host aphids and observing the cellular alterations of the apical germaria of ovarioles. Venom injection reproduced the same alterations observed in parasitized aphids, while injections of saline solution did not induce any detectable change. By 24h, the germarial cells of both venom-treated aphids and parasitized aphids showed the absence of the nucleolus and of electron-dense material around the nucleus, frequently referred to as "nuage material". By 48h more evident signs of degeneration were observed, suggesting the possible occurrence of apoptosis. The bioactive component of the venom was both heat- and protease-sensitive. The activity was found in purified fractions that were highly enriched in two proteins with an approximate molecular mass of 21kD and 36kD, respectively. These macromolecules are the most abundant components of A. ervi venom and, unlike many venom proteins of studied parasitic Hymenoptera, are not glycosylated and appear to be subunits of an oligomeric protein. The adaptive significance of host castration is discussed. PMID- 10802118 TI - Physiological aspects of multiple blood feeding in the malaria vector Anopheles tessellatus. AB - The malaria vector Anopheles tessellatus is able to take several blood meals in a gonotrophic cycle. The fecundity is largely dependent on the first blood meal and is not generally increased by subsequent blood meals during a gonotrophic cycle. Larval rearing densities influenced adult body size. There is an inverse relationship between wing length and larval rearing densities. Smaller mosquitoes produced from larvae reared at higher densities had reduced body reserves of protein, lipid and carbohydrates. At emergence, ovarian development in An. tessellatus is in the previtellogenic stage and it remained at this stage until the intake of a blood meal. The number of ovarian follicles is related to wing length and, irrespective of adult body size, An. tessellatus developed oocytes to maturity with a single blood meal. This is attributed to the availability of metabolic reserves above the threshold level required for further development of oocytes. Mosquitoes that took more than one blood meal had largely digested their previous blood meal and had ongoing vitellogenesis. Blood meals subsequent to the first one apparently contribute mainly to increasing metabolic reserves. The stimulus for a second and third blood meal in An. tessellatus appears to be completion of the digestion of the previous blood meal. There was no evidence that multiple blood meals taken in the first gonotrophic cycle influenced fecundity significantly in the second cycle. PMID- 10802119 TI - Estradiol-17beta in Bombyx mori: possible significance and its effect on silk production. AB - Although estrogen is well known as a vertebrate sex steroid, its presence in insects, including Bombyx mori, raises questions about its precise role in the physiology of insects. It was reported earlier that estradiol-17beta (E(2)) exerts a specific effect on silk-gland function in B. mori and that it may act in a nuclear-mediated way. To evaluate further the effect of E(2) on cocoon characters, larval growth and development, 1ug/g of E(2) was applied topically to the first and second day of fifth instar larvae. This resulted in a significant enhancement of cocoon characters, such as cocoon shell weight, silk filament length per cocoon, denier per filament and reelability of the cocoons, without any adverse effect on fecundity and hatchability. In the present study, E(2) levels in the haemolymph were quantified on different days of the fifth instar larvae and age-dependent changes in the endogenous E(2) titre have been demonstrated. These age-dependent variations in E(2) content coincide with physiological events occurring during the fifth instar. Such observations exclude the possibility of a dietary origin for E(2), as a sudden and sharp rise of the E(2) level in the haemolymph was observed on the 10th day of the fifth instar, preceded by a small increase on the ninth day after an eight-day feeding period. The increased level of estradiol in the haemolymph of larvae treated topically with E(2) indicates effective penetration of this hormone through the larval cuticle. Moreover, similar patterns of alteration of E(2) levels on different days of the fifth instar in both control and treated groups suggests the existence of some internal metabolic pathway in the silkworm body to regulate the hormone titre. Thus, the present investigation offers a system for investigating the unique function of E(2) in B. mori and offers potential for improvement of silk production. PMID- 10802120 TI - Aging and the human vestibular nuclei: morphometric analysis. AB - The data concerning the effects of age on the brainstem are scarce and few works are devoted to the human vestibular nuclear complex. The study of the effects of aging in the vestibular nuclei could have clinical interest due to the high prevalence of balance control and gait problems in the elderly. We have used in this work eight human brainstems of different ages sectioned and stained by the formaldehyde-thionin technique. The neuron's profiles were drawn with a camera lucida and Abercrombie's method was used to estimate the total number of neurons. The test of Kolmogorov-Smirnov with the correction of Lilliefors was used to evaluate the fit of our data to a normal distribution and a regression analysis was done to determine if the variation of our data with age was statistically significant. Aging does not affect the volume or length of the vestibular nuclear complex. Our results clearly show that neuronal loss occurs with aging in the descending (DVN), medial (MVN), and lateral (LVN) vestibular nuclei, but not in the superior (SVN). There are changes in the proportions of neurons of different sizes but they are not statistically significant. The neuronal loss could be related with the problems that elderly people have to compensate unilateral vestibular lesions and the alterations of the vestibulospinal reflexes. The preservation of SVN neurons can explain why vestibulo-ocular reflexes are compensated after unilateral vestibular injuries. PMID- 10802121 TI - Ageing and melatonin influence on in vitro gonadotropins and prolactin secretion from pituitary and median eminence. AB - The effect of ageing and/or melatonin (MEL) on in vitro gonadotropins, luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) release and tissue content from pituitary and median eminence (ME) were investigated. Gonadotropins and PRL basal release (I-1) from hemipituitaries of young cyclic-rats was decreased by MEL to levels shown in old acyclic rats. Pituitary tissue content of LH and PRL were not affected by ageing or MEL treatment. However, pituitary FSH tissue content was decreased by ageing and MEL, suggesting a different regulatory mechanism. MEL inhibitory influence on pituitary hormones is mainly exerted on the secretory process. This effect is only exerted in young rats. ME LH and PRL release and content were significantly lower than in pituitary. However, FSH release and content in ME showed values similar to those found in the pituitary. This study confirms that the functional capacities of pituitary gland and ME are maintained during reproductive senescence. PMID- 10802122 TI - An in vitro model of ischemic-like stress in retinal pigmented epithelium cells: protective effects of antioxidants. AB - We have developed a model of in vitro cell oxidative stress in bovine retinal pigment epithelium cells exposed to a ischemia-like condition obtained by interference with glucose utilization through both oxidative phosphorylation and glycolysis. This resulted in a statistically significant decrease of the intracellular ATP levels, which reflects a bioenergetic decline similar to that associated with mitochondrial damage or loss in normal post-mitotic cells aging in vivo. This new model of cellular oxygen stress seems adequate for investigation of the protective action of antioxidants, in agreement with our finding of a statistically significant increase in the ATP levels over the values of the non-treated samples in retinal pigment epithelium cells exposed to the above oxygen stress in medium supplemented with 300 microM vitamin C or 10 mM N acetylcysteine. PMID- 10802123 TI - Learning deficiency and alterations in acetylcholine receptors and protein kinase C in the brain of senescence-accelerated mouse (SAM)-P10. AB - The senescence-accelerated mouse (SAM) is known to be a murine model for accelerated aging. A novel inbred SAMP10 has shown age-related brain atrophy and learning deficiency. In the present study, we investigated the changes in learning ability and in ligand binding with muscarinic acetylcholine (mACh) receptors, alpha adrenoceptors and protein kinase C in SAMP10. In Morris's water maze task, in a control strain of SAMR1 at 9 months, the escape latency and path length decreased with increasing trial days, in contrast, escape latency and path length did not decrease in SAMP10. These results indicate that SAMP10 exhibits learning deficiency. The ligand binding activity of mACh receptors decreased in the hippocampus of SAMP10 and the protein kinase C level in the hippocampus of SAMP10 was lower than that of SAMR1. On the other hand, there was no significant difference between SAMR1 and SAMP10 regarding ligand binding activity of alpha(1) and alpha(2) adrenoceptors. Thus, a reduction of mACh receptors and protein kinase C in the brain seems to underlie dysfunction of learning and memory in SAMP10. PMID- 10802124 TI - For debate: defective mitochondria, free radicals, cell death, aging-reality or myth-ochondria? AB - As both experimental evidence and theoretical considerations may suggest that free radicals and mitochondria might be associated as key factors in aging, these organelles have been implicated in various versions of the free radical theory of aging. However, except for a few cases, no evidence for a death process specifically activated in respiratory defective cells could be found in patients with a mitochondrial disorder, including those harboring high levels of mutant mtDNA associated with profound respiratory chain deficiencies. This and more recent evidence suggest that damages produced by free-radicals endogenously generated in the mitochondria result in a distinctive biochemical profile, only occur under exceptional conditions and that a dysfunction of the respiratory chain does not cause opening of the permeability transition pore and is not sufficient per se to trigger massive entrance of cells into death processes, neither apoptosis nor necrosis. Therefore, defective mitochondria and their particular genome, should not be considered as a major and primary source of free radicals either leading cells into a death cascade or resulting in an accelerated aging process. PMID- 10802125 TI - An hydroalcoholic extract of Curcuma longa lowers the abnormally high values of human-plasma fibrinogen. PMID- 10802126 TI - Prevention of recurrent depressive episodes with milnacipran: consequences on quality of life. AB - BACKGROUND: Depression is now seen as a chronic disabling condition that spans the patient's lifetime and creates significant medical, economic and quality of life consequences. METHODS: 500 depressed patients were treated with milnacipran for 6 months. A total of 214 recovered patients were randomised to receive either milnacipran (50 mg bid) or a matching placebo for a 1-year recurrence prevention phase. Recurrence rate was the primary criteria; quality of life (QoL) consequences were evaluated through a shortened version of the French Sickness Impact Profile (SIP), the Depression Impact Profile (DIP). RESULTS: Milnacipran demonstrated its ability to reduce recurrences (Odds-Ratio=1.5; P<0.05), with a more marked effect in recovered patients with few residual symptoms (Odds Ratio=3.0). Responders who continued treatment with milnacipran had a dramatic improvement in their quality of life, although patients with residual symptoms still experienced some deterioration in their QoL (recreation, emotional, social, alertness and home assistance scores). Even recovered patients having zero scores on the Hamilton Depression Rating Scale-21 items (HDRS) had some QoL deterioration. The DIP emotional score was found to be more predictive of recurrence than the HDRS. Overall, the QoL was improved for those in the milnacipran group in comparison to placebo on the mobility, communication, psycho social and total scores. LIMITATIONS: The QoL evaluation was a secondary criteria; no a priori sample size estimate was performed. The choice of a generic QoL instrument might have reduced the sensitivity to clinical changes in depression. CONCLUSIONS: Prevention of recurrence in MDD with milnacipran contributes to an improvement in the QoL. PMID- 10802127 TI - Symptoms of anxiety and depression among mothers of pre-school children: effect of chronic strain related to children and child care-taking. AB - BACKGROUND: Factors that affect maternal mental health were studied when the children were 30 and 50 months old, and changes in the importance of these factors over time were analyzed. A specific aim was to elucidate the role of chronic strain related to children and child care-taking. This study follows up previous work on the influence of social class, strain and social support on maternal mental distress when the children were 18 months old. METHODS: The sample is population based, and 1,081 parents were invited to fill out questionnaires. Maternal mental distress was measured by the Hopkins Symptom Checklist (SCL-25). Multiple regression analyses were conducted at each time point and chi-square tests were used to analyze the changes between the estimated regression coefficients over time. RESULTS: Chronic strain related to children and child care-taking consistently predicted maternal mental distress. Among the specific child related strains, problems with child care-taking were significantly associated with maternal symptom levels at all time points. The importance of two specific child problem behaviors (activity level and the child being a worrier) on maternal mental health changed over time. LIMITATIONS: Conclusions about causality can not be drawn based on cross-sectional analyses. The self-report measures used here may be biased by the current mood state. CONCLUSIONS: Problems with child care arrangements and combining work and child care-taking are predictive of maternal mental health when the children are 18, 30 and 50 months old. The risk and protective factors found here may have implications for prevention and intervention. PMID- 10802128 TI - Efficacy of intravenous citalopram compared with oral citalopram for severe depression. Safety and efficacy data from a double-blind, double-dummy trial. AB - BACKGROUND: Intravenous administration is often beneficial in the treatment of severely depressed patients. It is mainly the tri- and tetracyclic antidepressant drugs that can be administered intravenously. However, these drugs have a less favourable safety profile than newer antidepressants, such as the selective serotonin reuptake inhibitors (SSRIs). Citalopram is the only SSRI that is available in a formulation for infusion. This double-blind, randomised, multicentre trial was designed to compare the efficacy and tolerability of citalopram infusion (40 mg per day) and citalopram tablet (40 mg per day). METHODS: Patients were randomised to receive either placebo tablet plus citalopram infusion (the infusion group; n=135) or citalopram tablet plus placebo infusion (the tablet group; n=119). After receiving randomised treatment for eight days, all patients entered an open treatment phase, during which they received oral citalopram 40 mg per day for five weeks. RESULTS: Although there was no difference in Montgomery-Asberg Depression Rating Scale (MADRS) scores at the end of the randomised treatment period, by the end of the open treatment phase the reduction in MADRS scores was significantly greater in the infusion group than in the tablet group (p=0.015). The infusion group also showed superior efficacy in Clinical Global Impressions assessments. Citalopram was equally well tolerated in both treatment groups. CONCLUSIONS: This trial confirmed the efficacy of citalopram 40 mg per day, and clearly supports the use of citalopram infusion in the treatment of severely depressed, hospitalised patients. PMID- 10802129 TI - Negative association between T102C polymorphism at the 5-HT2A receptor gene and bipolar affective disorders in Singaporean Chinese. AB - BACKGROUND: Serotonergic system abnormalities have been implicated in the pathogenesis of bipolar affective disorders. The 5-hydroxytryptamine type 2A (5HTR2A) receptor gene located on chromosome 13 (13q14-21) can be considered as a candidate gene for bipolar affective disorder (BPAD). METHODS: Seventy-two patients with BPAD and 74 normal population controls were genotyped with restriction fragment length polymorphism (RFLP) in the 5HTR2A receptor gene. RESULTS: We report a negative association between 5HTR2A receptor gene and BPAD. The association was examined using a case-control design. Allele and genotype frequencies as well as homozygote-heterozygote distribution at the 5HTR2A receptor gene polymorphism were compared between the two groups. There were no significant differences in the allelic or genotype frequencies and the homozygote heterozygote distributions. LIMITATIONS: Patients were recruited from one hospital in Singapore. The case-control study design needs replication. CONCLUSION: Our finding indicates that the 5HTR2A receptor gene polymorphism is not a major factor in the genetic susceptibility to BPAD in Singaporean Chinese. PMID- 10802130 TI - Early and late onset bipolar disorders: two different forms of manic-depressive illness? AB - BACKGROUND: Conflicting results in genetic studies of bipolar disorders may be due to the clinical and genetic heterogeneity of the disease. Age at onset of bipolar disorders may be a key indicator for identifying more homogeneous clinical subtypes. We tested whether early onset and late onset bipolar illness represent two different forms of bipolar illness in terms of clinical features, comorbidity and familial risk. METHODS: Among a consecutively recruited sample of 210 bipolar patients, we compared early onset (n=58) and late onset (n=39) bipolar patients; the cut-off points were age at onset before 18 years and after 40 years for the two subgroups. The subgroups were compared by independent t tests and a contingency table by raw chi-square test. Morbid risk among first degree relatives was measured by the survival analysis method. RESULTS: The early onset group had the most severe form of bipolar disorder with more psychotic features (P=0.03), more mixed episodes (P=0.01), greater comorbidity with panic disorder (P=0.01) and poorer prophylactic lithium response (P=0.04). First degree relatives of early onset patients also had a higher risk of affective disorders (P=0.0002), and exhibit the more severe phenotype, i.e bipolar disorder. CONCLUSION: Our data suggest that early and late onset bipolar disorders differ in clinical expression and familial risk and may therefore be considered to be different subforms of manic-depressive illness. PMID- 10802131 TI - Prevalence of DSM IV anxiety and affective disorders in a pediatric population of asthmatic children and adolescents. AB - A series of 82 children and adolescents with moderate and severe persistent asthma was studied. Their psychopathological problems were compared to those of 82 healthy subjects, matched for age, sex and socio-economic status. The patients completed the Child Depression Inventory, an inventory of fears and anxiety (ECAP) and the Coopersmith Self Esteem Inventory. Parents of asthmatic children filled in the Child Behavior Check List to assess their social competence. The patients were examined with the revised Kiddie Schedule for Affective Disorders and Schizophrenia. There were more anxiety symptoms in the asthmatic group than in the control group. Asthmatics were not significantly more depressed than controls and their self-esteem was as good. We found 29 anxiety disorders, four affective disorders and four disruptive behavior disorders. Generalized anxiety disorder was the main diagnosis (n=24). The asthmatic subgroup presenting anxiety and affective disorders had poorer self esteem, fewer activities and worse social competence than other asthmatics and controls. Adolescents did not seem to have more emotional disturbances than younger patients. Girls did not have more DSM IV anxiety or affective disorders than boys. PMID- 10802132 TI - Four-year follow-up study of sertraline and fluvoxamine in long-term treatment of unipolar subjects with high recurrence rate. AB - We prolonged from 24 to 48 months a follow-up study of unipolar subjects with high recurrence rate treated with fluvoxamine (N=25) and sertraline (N=22). During the two-year additional period a significant risk of recurrences was observed during the third year of follow-up, without differences in the two long term therapy groups. During the fourth year no patients showed new episodes of illness. PMID- 10802133 TI - Perception of ill spouse and dyadic relationship in couples with affective disorder and those without. AB - BACKGROUND: This study compares the perception of spouses and the quality of the dyadic relationship of patients with severe affective disorders in remission with healthy couples. METHOD: The sample included spouses of patients between the ages of 20 and 65 who had been hospitalized with severe affective illness and who were currently in remission (depressive, n=23; bipolar, n=11) and a control group matched by socioeconomic status. Both groups completed three instruments measuring the quality of the dyadic relationship, attributed characteristics of the spouse and non-formal social support. RESULTS: The spouses of patients, as compared to spouses of controls, scored lower on consensus, unity and expressions of affection in their marital relationship, ranked their ill spouses lower on the positive qualities and higher on the negative qualities and reported receiving less emotional and practical support. LIMITATIONS: The limitations of the study are the small sample size and cross sectional design. CONCLUSIONS: The results suggest that severe affective disorders are associated with marital dysfunction, even during periods of symptom remission. PMID- 10802134 TI - Lower serum vitamin E concentrations in major depression. Another marker of lowered antioxidant defenses in that illness. AB - OBJECTIVE: Major depression is associated with defective antioxidant defenses. Vitamin E is the major fat soluble antioxidant in the body. The aim of the present study is to examine serum vitamin E concentrations in major depressed patients versus normal volunteers. METHOD: Serum vitamin E concentrations were measured in 26 healthy volunteers and 42 major depressed patients by means of HPLC. Since vitamin E is a fat soluble vitamin, and serum vitamin E concentrations are strongly related to these of low-density-lipoprotein cholesterol (LDL-C) and triglycerides, we have adjusted the results for possible differences in these lipids. The numbers of peripheral blood leukocytes were measured. RESULTS: Patients with major depression had significantly lower serum vitamin E concentrations than healthy controls. The area under the ROC (receiver operating characteristics) curve was 83%. There were significant and negative correlations between serum vitamin E and number of total leukocytes and neutrophils. CONCLUSIONS: Major depression is accompanied by significantly lower serum vitamin E concentrations, suggesting lower antioxidant defenses against lipid peroxidation. The results could, in part, explain previous findings, which suggest increased lipid peroxidation in major depression. PMID- 10802135 TI - Chromogenic plate assay distinguishing bacteriolytic from bacteriostatic activity of an antibiotic agent. AB - A solid agar plate assay was devised to discriminate bacteriolytic from bacteriostatic activity for a given antibacterial agent. The assay uses a bacterial culture harboring beta-galactosidase enzyme as reporter of cellular lysis. When a drop of bacteriolytic compound is placed on the agar, beta galactosidase is released from the bacteria to the external solid medium where it hydrolyzes X-Gal substrate analogue, developing a blue halo at the edge of the inhibition growth zone. The assay was successfully evaluated against several antibiotics with well-known mechanism of action. It was found that bacteriostatic compounds consistently did not display blue halo at the inhibition zone. PMID- 10802136 TI - A bioluminescent Escherichia coli auxotroph for use in an in vitro lysine availability assay. AB - Microbiological methods have been used to determine the amino acid availability of a variety of animal feed and human food protein sources. Growth of Escherichia coli auxotrophs have been shown to yield a consistent linear response to lysine concentration when compared to chemical measures. Extent of total growth of E. coli lysine mutant (American Type Culture Collection #23812) when measured as optical density (OD) displays a lysine-dependent growth response that can be used to estimate lysine in feed proteins. However, typical OD-based growth studies for amino acid quantitation using the mutant may require anywhere from 12 to over 40 h. To develop an improved rapid method for lysine quantitation in protein sources, the plasmid pJHD500 carrying genes that encode for expression of bioluminescence and ampicillin resistance was transformed into the E. coli mutant by electroporation (set at 1.80 kV). The luminescence measured during early exponential growth allowed detectable differentiation of lysine concentration in the media in 4 h. When the luminescence method was compared with the conventional optical density lysine growth assay, the correlation coefficient was 0.989. Lysine availability valued for enzymatically hydrolyzed protein sources were comparable with availability measures using animal methods for lysine availability. This research shows potential applications for more rapid quantitative measurement of bioavailable lysine. PMID- 10802137 TI - BASIC program for reduction of data from community-level physiological profiling using biolog microplates: rationale and critical interpretation of data. AB - A BASIC program is offered that reduces data resulting from mixed-species inoculations into Biolog microplates. The procedures of the program are supported by a critical review of the literature relating to Biolog data reduction. The availability of standardized, accelerated data reduction protocols will facilitate study comparisons and allow efficient evaluation of new data reduction approaches. PMID- 10802138 TI - Modification of the phosphoketolase assay for rapid identification of bifidobacteria. AB - The phosphoketolase assay is commonly used as a definitive criterion for identification of bifidobacteria. A limitation of the assay is the time-consuming process of cell disruption, either by use of the French Pressure Cell or by sonication. We have replaced the time consuming cell disruption process with a more rapid cell membrane disruption process by pretreating cells with the detergent hexadecyltrimethylammonium bromide (cetrimonium bromide, CTAB). The effect of no pretreatment, sonication or the addition of CTAB (0.45 mg/ml) on color development in the phosphoketolase assay was tested using pure cultures of bifidobacteria and lactobacilli. No phosphoketolase activity was observed with bifidobacterial cultures without cell disruption or with lactobicilli that had undergone cell disruption. All bifidobacterial cultures gave a similar color formation whether sonication or CTAB addition was used to disrupt cells. Use of CTAB to disrupt cell membranes is an effective alternative to the time consuming traditional cell disruption procedures and increases the number of cultures that can be simultaneously assayed and presumptively identified using the phosphoketolase assay. PMID- 10802139 TI - Dot assay for determining adhesive interactions between yeasts and bacteria under controlled hydrodynamic conditions. AB - Candida belongs to the normal human microflora and are found adhering to a number of human body tissues as well as to a variety of biomaterials implants. Often, yeasts adhere in association with bacteria, but to date there is no definitive assay to investigate adhesive interactions between yeasts and bacteria adhering on surfaces. Although we recently described the use of a parallel plate flow chamber to this purpose [Millsap, K.W., Bos, R., Van der Mei, H. C., Busscher, H.J., 1998. Adhesive interactions between medically important yeasts and bacteria. FEMS Microbiol. Rev. 21, 321-336], the method was slow and evaluation of a large number of strains showed major biological variation between experiments. Here, we describe a new assay for the simultaneous determination of the adhesive interactions between yeasts and different bacterial strains on a surface under controlled hydrodynamic conditions. On an acrylic surface, the presence of adhering bacteria suppressed adhesion of Candida albicans ATCC 10261 to various degrees, depending on the bacterial strain involved. Suppression of C. albicans ATCC 10261 adhesion was strongest by Actinomyces naeslundii T14V-J1, while adhering Streptococcus gordonii NCTC 7869 caused the weakest suppression of yeast adhesion. When adhering yeasts and bacteria were challenged with the high detachment force of a passing liquid-air interface, the majority of the yeasts detached, while C. albicans adhering on the control, bare polymethylmethacrylate surface formed aggregates. Summarizing, this study presents a new method to determine suggested adhesive interactions between yeasts and adhering bacteria under controlled hydrodynamic conditions. However, the results seem to indicate that these adhesive interactions may well not exist, but that instead different bacterial strains have varying abilities to discourage yeast adhesion. PMID- 10802140 TI - Flowcell culture of Porphyromonas gingivalis biofilms under anaerobic conditions. AB - We have developed an anaerobic biofilm culture system. The system is inexpensive, simple to use and, unlike an anaerobic glovebox, requires no dedicated space. As a test of the system, Porphyromonas gingivalis was cultured under low oxygen (1-2 ppm) and under anaerobic conditions ( 18:0 > 16:1 = 18:2 > 16:0 = 14:0 > 20:4 and differentiation did not affect the relative amounts of these fatty acids within individual phospholipids. When cells were incubated with [1,3-(3)H]glycerol, radioactivity incorporated into phosphatidylethanolamine was elevated 5.8-fold, whereas radioactivity incorporated into phosphatidylcholine was unaltered. Ethanolaminephosphotransferase, cholinephosphotransferase and membrane CTP:phosphocholine cytidylyltransferase activities were elevated in differentiated cells compared to undifferentiated cells, whereas membrane and cytosolic phospholipase A2 activities were unaltered. Lysophosphatidylethanolamine acyltransferase activities were elevated 2.4-fold (P<0.05). Lysophosphatidylcholine acyltransferase, monolysocardiolipin acyltransferase, acyl-Coenzyme A synthetase and acyl-Coenzyme A hydrolase activities were unaltered in differentiated cells compared to undifferentiated cells. We postulate that during cardiac cell differentiation, the observed elevation in lysophosphatidylethanolamine acyltransferase activity accompanies the elevation in phosphatidylethanolamine mass, possibly to maintain the fatty acyl composition of this phospholipid within the membrane. PMID- 10802245 TI - Plasma thiols inhibit hemin-dependent oxidation of human low-density lipoprotein. AB - Oxidative modification of human low-density lipoprotein (LDL) renders it atherogenic. Previous studies demonstrated that plasma thiols promote oxidation of LDL by free ferric iron (Fe3+). The current study investigated effects of plasma thiols on oxidation of LDL by hemin, a physiological Fe3+-protoporphyrin IX complex thought to be capable of initiating LDL oxidation in vivo. In contrast to free Fe3+ which is incapable of oxidizing LDL in the absence of an exogenous reductant, hemin readily promoted LDL oxidation. During incubation of LDL (0.2 mg of protein/ml) with hemin (10 microM) at 37 degrees C for 6 h, thiobarbituric acid-reactive substances (TBARS), a marker of lipid oxidation, increased from 0.3 (+/-0.1) nmol/mg of LDL protein to a maximal concentration of 45.8 (+/-5.2) nmol/mg of LDL protein. Under the same experimental conditions, lipid-conjugated dienes, another marker of lipid oxidation, increased from non-detectable to near maximal levels of 78-187 nmol/mg of LDL protein, and lipoprotein polyunsaturated fatty acyl-containing cholesteryl ester content decreased to 15-36% of that present in native (i.e. unoxidized) LDL. Continued incubation of LDL with hemin for up to 24 h resulted in no further significant alterations in lipoprotein levels of TBARS, lipid-conjugated dienes, and cholesteryl esters. In addition to these chemical modifications indicative of lipoprotein oxidation, agarose gel electrophoretic analysis indicated that exposure of LDL to hemin resulted in conversion of the lipoprotein to an atherogenic form as evidenced by its increased anodic electrophoretic mobility. Addition of physiological concentrations of plasma thiols (either cysteine, homocysteine or reduced glutathione; 1-100 microM, each) inhibited hemin-mediated oxidation of LDL. Thus, whereas the maximal TBARS concentration was achieved following 6 h of incubation of LDL with hemin alone, addition of thiol extended the time required to attain maximal TBARS concentration to > or = 12 h. Similar antioxidant effects of thiols on formation of lipid-conjugated dienes, loss of cholesteryl esters, and lipoprotein anodic electrophoretic mobility were also observed. However, all thiols were not equally effective at inhibiting hemin-dependent LDL oxidation. Thus, whereas reduced glutathione was most effective at inhibiting hemin dependent LDL oxidation, an intermediate effect was observed for homocysteine, and cysteine was least effective. The inhibition of hemin-mediated LDL oxidation by plasma thiols reported here confirms a previous observation that, under certain conditions, thiols can function as antioxidants, but contrasts with the previously documented pro-oxidant effect of the same thiols on oxidation of LDL by free Fe3+. These contrasting effects of plasma thiols on hemin- and free Fe3+ mediated LDL oxidation indicate that, in vivo, the ability of thiols to function as either anti- or pro-oxidants during LDL oxidation may, at least in part, be determined by the type of oxidant stress to which the lipoprotein is exposed. PMID- 10802246 TI - Comparative effects of perilla and fish oils on the activity and gene expression of fatty acid oxidation enzymes in rat liver. AB - The activity and mRNA level of hepatic enzymes in fatty acid oxidation and synthesis were compared in rats fed diets containing either 15% saturated fat (palm oil), safflower oil rich in linoleic acid, perilla oil rich in alpha linolenic acid or fish oil rich in eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) for 15 days. The mitochondrial fatty acid oxidation rate was 50% higher in rats fed perilla and fish oils than in the other groups. Perilla and fish oils compared to palm and safflower oils approximately doubled and more than tripled, respectively, peroxisomal fatty acid oxidation rate. Compared to palm and safflower oil, both perilla and fish oils caused a 50% increase in carnitine palmitoyltransferase I activity. Dietary fats rich in n-3 fatty acids also increased the activity of other fatty acid oxidation enzymes except for 3 hydroxyacyl-CoA dehydrogenase. The extent of the increase was greater with fish oil than with perilla oil. Interestingly, both perilla and fish oils decreased the activity of 3-hydroxyacyl-CoA dehydrogenase measured using short- and medium chain substrates. Compared to palm and safflower oils, perilla and fish oils increased the mRNA level of many mitochondrial and peroxisomal enzymes. Increases were generally greater with fish oil than with perilla oil. Fatty acid synthase, glucose-6-phosphate dehydrogenase, and pyruvate kinase activity and mRNA level were higher in rats fed palm oil than in the other groups. Among rats fed polyunsaturated fats, activities and mRNA levels of these enzymes were lower in rats fed fish oil than in the animals fed perilla and safflower oils. The values were comparable between the latter two groups. Safflower and fish oils but not perilla oil, compared to palm oil, also decreased malic enzyme activity and mRNA level. Examination of the fatty acid composition of hepatic phospholipid indicated that dietary alpha-linolenic acid is effectively desaturated and elongated to form EPA and DHA. Dietary perilla oil and fish oil therefore exert similar physiological activity in modulating hepatic fatty acid oxidation, but these dietary fats considerably differ in affecting fatty acid synthesis. PMID- 10802247 TI - Lipid peroxidation and glutathione system in hyperlipemic rabbits: influence of olive oil administration. AB - We studied the effect of supplementation (10% w/w) of a hyperlipemic diet (1% cholesterol) with olive oil (OLIV) for 6 weeks in four groups of 10 rabbits each. At the end of this period, we determined lipid peroxidation, glutathione content, and glutathione peroxidase, reductase and transferase activities in liver, brain, heart, aorta and platelets. The atherogenic diet increased tissue lipid peroxidation and decreased the protective antioxidant effect of glutathione. Dietary supplementation with olive oil reduced tissue lipid peroxidation by 71.6% in liver, 20.3% in brain, 84.5% in heart, 63.6% in aorta, 72% in platelets. The ratios total/oxidized glutathione were increased in all tissues (49% in liver, 48% in brain, 45% in heart, 83% in aorta, 70% in platelets). Olive oil increased glutathione peroxidase and transferase activities in all tissues. We conclude that in rabbits made hyperlipemic with a diet rich in saturated fatty acids, olive oil decreased tissue oxidative stress. PMID- 10802248 TI - Palmitate-induced apoptosis in cardiomyocytes is mediated through alterations in mitochondria: prevention by cyclosporin A. AB - Palmitate, a C16 fatty acid found in high concentrations in the blood in acute myocardial infarction, induces apoptotic cell death. To more completely define the nature and mechanism underlying palmitate-induced cell death, cardiomyocytes were cultured from embryonic chick heart and were treated with palmitate. Concentration-dependent loss of cell viability was established by loss of the ability of palmitate-treated cells to exclude propidium iodide (PI), metabolize 3 [4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and retain fluorescein diacetate (FDA). Dual staining with PI and FDA and subsequent analysis by FACS established that palmitate-induced cell death was predominantly necrosis whereas apoptosis occurred in 13% of all dead cells. The low proportion of palmitate-induced apoptosis was confirmed by evaluation of the DNA content or PI fluorescent staining of the DNA of permeabilized cardiomyocytes. A critical role for mitochondria in the pathogenesis of palmitate-induced cell death was demonstrated, for the first time, based on palmitate-induced reduction of mitochondrial activity as assessed by the mitochondrial-selective dye chloromethyl-X-Rosamine and the presence of a greater amount of the mitochondrial marker cytochrome C in the cytosol of palmitate-treated cardiomyocytes than in control cells. Further, cyclosporin that inhibits the development of mitochondrial transition pores blocked palmitate-induced alteration in mitochondrial function and palmitate-induced cell death. We further demonstrated the selectivity of cyclosporin A for the prevention of apoptotic cell death in the heart as there was no alteration in necrotic cell death produced by palmitate with cyclosporin pretreatment. Our data demonstrate the nature of palmitate induced cell death in cardiomyocytes (both apoptotic and necrotic), propose a mitochondrial basis for its pathogenesis and show that cyclosporin A prevents palmitate-induced apoptotic cardiomyocyte cell death. PMID- 10802249 TI - Egg yolk lipoproteins as substrates for lipases. AB - Egg yolk emulsions containing phospholipids (about 31%, w/w) are classically used as substrates for measuring phospholipase A2 activity using the pH-stat method. Here we investigated the susceptibility of egg yolk lipoproteins to lipolysis by various highly purified lipases of animal or microbial origin. Egg yolk lipoproteins, which contain up to 65% triacylglycerols, were found to be effective substrates for all the lipases tested. The specific activities measured on egg yolk lipoproteins using the pH-stat technique were found to be 8000, 1000, 1250 and 1700 U/mg in the case of human pancreatic lipase, horse pancreatic lipase, porcine pancreatic lipase and Humicola lanuginosa lipase, respectively. No activity was detected in the absence of colipase with any of the pancreatic lipases tested. Consequently, the classical egg yolk assay cannot be considered as a specific phospholipase A2 assay. PMID- 10802252 TI - Discussion PMID- 10802251 TI - Importance, morbidity, and mortality of pneumonia in the surgical intensive care unit. AB - Surgical patients are at high risk to develop nosocomial pneumonia, although an accurate diagnosis is difficult to make. Staphylococcus aureus and Pseudomonas aeruginosa are the most common pathogens, but Acinetobacteris emerging as an important pathogen. Because affected patients are often critically ill with multisystem pathology, it can be difficult to ascribe morbidity or mortality directly to the infection. PMID- 10802253 TI - Identification of patients at highest risk for ventilator-associated pneumonia in the surgical intensive care unit. AB - Ventilator-acquired pneumonia (VAP) is a major problem for patients admitted to the surgical intensive care unit mechanically ventilated. Recently, we have identified both clinical and immunologic factors associated with the development of VAP. The clinical risk factors are associated with the severity of the injury and the length of mechanical ventilation. The immunologic risk factors are associated with the local lung inflammatory response that is unchecked and affects local cell function. The combination of the severity of injury, the length of mechanical ventilation, and the failure to "auto-regulate" the lung response places the host at risk of VAP. In the next millennium, if we are to make significant advances in the management of VAP, we will need to understand the pathophysiology of the disease process. Then we can develop preventive strategies that will reduce the morbidity and the associated cost of VAP. PMID- 10802254 TI - Discussion PMID- 10802255 TI - Prevention of ventilator-associated pneumonia AB - Ventilator-associated pneumonia (VAP) remains a common complication within the intensive care unit. Despite identification of the patients at highest risk for developing VAP, the actual incidence appears to be unchanged over the past two decades. Colonization of the oral pharynx with pathogenic organisms and ongoing aspiration appear to be necessary in the development of VAP. Thus, prevention strategies targeted at interrupting these factors should decrease VAP. In the few randomized prospective studies in the literature, both continuous aspiration of the subglottic space and the prevention colonization of the oral pharynx have been proven successful in decreasing VAP. The main problem in interpreting these results is that they have been generally single reports and need to be replicated in larger multicenter trials. Studies of "early" tracheostomy have been unable to define both the optimal timing of tracheostomy and its effect in decreasing VAP. Other modalities, such as rotating or kinetic beds, early bronchoscopy, and changes in ventilator management, have not been shown to be useful. PMID- 10802256 TI - Discussion PMID- 10802257 TI - Empiric therapy for pneumonia in the surgical intensive care unit. AB - Empiri c therapy of ventilator-associated pneumonia (VAP) in surgical patients should be based on intensive care unit (ICU)-specific surveillance data, because microbial flora patterns vary widely between geographic regions as well as within hospitals. Surgical ICUs have higher VAP rates than other units. Data from the National Nosocomial Infection Surveillance (NNIS) System report Pseudomonas aeruginosa and Staphylococcus aureus to be the most frequent isolates (each 17.4%). Data from the NNIS documents high resistance patterns in ICUs compared with hospitals at large, as well as unit-specific patterns. VAP risk factors for surgical patients include thoracoabdominal surgery, altered level of consciousness, advanced age, diabetes mellitus, malnutrition, chronic obstructive pulmonary disease, and prior antibiotic administration. Promising prevention strategies include restricting ventilator circuit changes, in-line heat moisture exchange filters, semi-recumbant positioning, and continuous subglottic aspiration. Pharmacodynamics should be considered when choosing antibiotic regimens. Postantibiotic effect and time-dependent versus concentration-dependent killing should be studied in clinical trials. Current guidelines for choosing regimens have been well developed by the American Thoracic Society. PMID- 10802258 TI - Discussion PMID- 10802259 TI - Diagnosis of acute respiratory distress syndrome and differentiation from ventilator-associated pneumonia. AB - Acute respiratory distress syndrome (ARDS) remains a significant cause of morbidity and mortality in surgical intensive care unit patients. A relatively predictable sequence of pathophysiologic events occur in the lung, which involve inflammatory mediators and neutrophils. The characteristic radiographic findings of ARDS may mimic ventilator-associated pneumonia (VAP), making the diagnosis of VAP difficult. The standard clinical criteria of fever, leukocytosis, purulent sputum, and infiltrate on chest radiograph are not specific for pneumonia in the surgical patient. The use of bronchoscopy with bronchoalveolar lavage and quantitative cultures can differentiate local and systemic inflammatory response to injury or blood loss from invasive bacterial pneumonia. Basing antibiotic therapy solely on the results of quantitative cultures is safe, because quantitative cultures identify VAP in less than half the patients with clinical evidence of pneumonia. Empiric therapy should be based on the microbiology of the intensive care unit rather than the results of the Gram stain. PMID- 10802260 TI - Discussion PMID- 10802261 TI - Multifactorial analyses in the diagnosis of pneumonia arising in the surgical intensive care unit. AB - The diagnosis of ventilator-associated pneumonia in the surgical intensive care unit continues to be problematic. The majority of intensive care units use clinical criteria based on chest x-ray; fever; leukocytosis; alterations in the pulse oximeter observations; the need to alter modes and amounts of ventilatory support; and more specific microbiologic studies, such as appropriate sputum, Gram stain, and culture to identify pneumonia. Diagnosing pneumonia based on clinical criteria alone is often difficult and inaccurate, which may lead to inappropriate use and choice of antibiotics. Invasive diagnostic techniques, such as protected specimen brush and bronchoalveolar lavage, provide an important microbiologic diagnosis. However, the cost and inconvenience limit broad usage. Furthermore, those results that return positive are often too late to dictate the need for, or direction of, therapy. Our use of a "pneumonia grid" may help identify patients likely to have a poor outcome. Until a readily available and cost-effective diagnostic study for pneumonia is developed, clinical criteria remain vital in routine practice. PMID- 10802262 TI - Pneumonia in the surgical patient: duration of therapy and does the organism matter? AB - Pneumonia is a frequent complication in the critically ill surgical or trauma patient. Despite its common occurrence and increased attention recently, many basic issues, such as how long to treat and whether the exact causative organism even matters, remain unanswered. Currently, cessation of treatment is based on clinical response, although some data suggest that both the microbiologic and immunologic response within the lung are also important. A thorough knowledge of the likely causative organisms for both early and late pneumonia should allow safe and efficacious empiric antibiotic selection. PMID- 10802263 TI - Discussion PMID- 10802264 TI - Immunologic responses to pneumonia. AB - Pneumonia in the critically ill surgical patient often results from the bombardment of a previously normal pulmonary system with therapeutic foreign bodies, hospital pathogens, and impairment of the host defenses. Despite its long history as a significant clinical problem, a woefully inadequate amount of study has been directed toward therapy. We created an experimental model of a differential pulmonary infection using a strain of Klebsiella pneumoniae. We then compared the progressively affected pneumonic process versus the normal parenchyma. We measured neutrophil and monocyte complement antibody receptor expression and monocyte and macrophage class II major histocompatibility antigens (HLA-DR) via percent of cells and mean fluorescent intensity outcomes from flow cytometry. The main difference between infected versus noninfected tissues was monocyte DR expression, which was consistently depressed in cells from infected parenchyma. What follows is a discussion of the implications of this work as well as other work in the immunology of pneumonia and cytokine expression. Possible therapeutic modalities are included. PMID- 10802265 TI - Role of aztreonam in the treatment of nosocomial pneumonia in the critically ill surgical patient. AB - In 1995 the American Thoracic Society issued an official consensus statement on the treatment of hospital-acquired pneumonia (HAP). Classes of antimicrobials included in the list of antimicrobials deemed to be suitable for the empiric treatment of severe HAP were the aminoglycosides, quinolones, antipseudomonal penicillins, carbapenems, and beta-lactam/beta-lactamase inhibitor combinations. Aztreonam, a monobactam, was also listed and is unique among these agents based on its spectrum of activity being limited to the gram-negative bacillary bacteria combined with an excellent safety profile. This review focuses on the role of aztreonam in the treatment of nosocomial pneumonia in the critically ill patient.A review of the literature was performed using PubMed and secondary literature sources as to the clinical efficacy of aztreonam in the treatment of lower respiratory tract infections as well as its pharmacokinetic and safety profiles. An analysis of aztreonam's potential pharmacoeconomic advantages compared with other agents was also performed. Numerous studies have documented that aztreonam has effectiveness that is equal or superior to that of other suitable antibiotics in the treatment of nosocomial pneumonia. Its excellent safety profile makes it a particularly attractive agent compared with the aminoglycosides. Considering the potential costs of bacterial resistance from the use of broader-spectrum alternatives, a case can be made that aztreonam is a pharmacoeconomically sound choice as well. PMID- 10802266 TI - Discussion PMID- 10802267 TI - Pharmacoeconomics of pneumonia. AB - Because diagnosis and treatment are so intimately linked, the pharmacoeconomics of treatment of ventilator-associated pneumonia (VAP) is impossible to discuss without discussing the cost-effectiveness of VAP diagnosis. The cost of VAP treatment is more complex than simply drug acquisition and administration costs. The critical factor in cost-effective therapy is the avoidance of inappropriate or ineffective therapy. The second most important benefit of a more accurate diagnostic strategy, such as the use of quantitative cultures, is the ability either to stop or to withhold antibiotics if the quantitative culture is negative. Therefore, the benefit of any diagnostic strategy must be evaluated principally from the aspect of these resultant changes in management. Reassurance or concern about an alternative site of infection or cause of fever will also add to the benefit or cost of more accurate diagnosis of VAP. The baseline antibiotic treatment strategy of the specific intensive care unit (ICU) will determine, to a large degree, the cost of antibiotics and the efficacy of empiric regimens. In the final analysis, pharmacy costs and cost of diagnostic testing for VAP must be based on outcome analysis, including comparison of the more expensive aspects of care, such as mortality, length of mechanical ventilation, and length of ICU stay. PMID- 10802268 TI - Single versus combination antimicrobial therapy for ventilator-associated pneumonia. AB - The appropriate selection of definitive antimicrobial therapy is a necessary component of the overall treatment for ventilator-associated pneumonia. When possible, single-agent therapy is preferable. A combination of antibiotics is necessary to treat multiple organisms not susceptible to a single appropriate antibiotic and when antibiotic-resistant gram-negative bacteria are present. Treatment failure is more commonly the result of persistent pneumonia and the development of antibiotic resistance than to recurrence after successful antimicrobial therapy. The duration of treatment will vary depending on the severity of the underlying illness and the pneumonic process. PMID- 10802270 TI - Discussion PMID- 10802269 TI - Complications of nosocomial pneumonia in the surgical patient. AB - Nosocomial pneumonia is a leading cause of morbidity and mortality in the surgical and trauma patient. Inadequate treatment can lead to the complications of acute respiratory distress syndrome (ARDS), empyema, and lung abscess. The prevention and treatment of these complications revolve around several key principles. Complete treatment of pneumonia requires appropriate antimicrobial therapy, as well as mechanical pulmonary hygiene and proper airway management. Despite advances in treatment of pneumonia, complications arise necessitating treatment. This article reviews the treatment of ARDS, empyema, and lung abscesses. In particular, the many options for treatment of empyema are discussed in detail. Additionally, the treatment of pulmonary contusion and hemopneumothorax in the trauma patient is discussed. The understanding of sound treatment principles in the critically ill postsurgical patient helps prevent complicated or recurrent pneumonia and allows the surgeon to intervene effectively when such complications occur. PMID- 10802271 TI - Low-dose oral contraceptives and bone mineral density: an evidence-based analysis. AB - We reviewed studies of the association of oral contraceptive (OC) use and bone mineral density (BMD). We limited the review to studies of women using low-dose oral contraceptives and that measured BMD by bone densitometry. A total of 13 studies met the inclusion criteria. Nine of these showed a positive effect of OC use on BMD, and four did not show an association. However, none of the studies showed a decrease in BMD with OC use. We classified the level of evidence from each study according to the guidelines of the US Preventive Services Task Force. The level of evidence supporting a positive association between OC use and increased BMD is II-1. There is fair evidence (Category B) to support the position that OC use has a favorable effect on BMD. We made suggestions for a study design that could yield Level I evidence. PMID- 10802272 TI - Norplant use among urban minority women in the United States. AB - The aim of this study was to develop a profile of urban American women who chose Norplant(R), determine factors associated with retention and early termination of implants, and to determine reasons for early removal. A total of 197 adult black and Hispanic women who had Norplant inserted were followed prospectively for up to 5 years. Interval and cumulative termination rates were calculated. Data were stratified by race and analyzed to include lost-to-follow-up (LTFU) subjects. Multivariate survival analysis was used to determine variables independently associated with termination. Cumulative continuation rates were 68% after year 1 and 13% after year 4. Significant predictors of retention included black race and lower parity. Probability of early termination increased with higher parity and Hispanic race. For black subjects, recent use of hormonal contraception was a predictor of retention. Menstrual changes and weight concerns were common reasons for removal. The Norplant 1-year continuation rate is lower than previously reported, but is higher than reported for oral contraceptives and Depo Provera(R). Future studies should stratify by age, race, and parity, and use standardized terminology to report intervals of use. PMID- 10802273 TI - Performance of the frameless GyneFix and the TCu380A IUDs in a 3-year multicenter, randomized, comparative trial in parous women. AB - This study was conducted to evaluate a new and improved inserter (GyneFix) for the anchoring of the Frameless IUD in the uterine cavity. Previous studies conducted with a prototype inserter (Flexigard) did not show fully the advantages of the new anchoring concept because of the shortcomings of the Flexigard inserter and the complexity of the insertion technique. The GyneFix IUD was compared with the TCu380A IUD in six centers in China in approximately 300 women in each group. Only parous women were included in the study. The data from this 3 year, ongoing study demonstrate that the shortcomings of the inserter have been corrected, resulting in better performance and a much reduced rate of failed insertion/expulsion of the frameless and anchored device. The cumulative expulsion rate with the GyneFix IUD was 3.0 at 3 years (annual rates 2.67, 0.33, and 0.0, respectively) compared with a cumulative expulsion rate of 7.38 at 3 years with TCu380A (annual rates 4.63, 1.76, and 1.04, respectively). This difference is statistically significant. The majority of the expulsions with the anchored IUD occurred early in the study, indicating improper anchoring technique. The study also shows that both devices are highly efficacious. No pregnancies occurred with the GyneFix IUD (cumulative pregnancy rate 0.0 at 3 years) versus one pregnancy with TCu380A (cumulative pregnancy rate 0.34 at 3 years). The total use-related discontinuation rate at 3 years was significantly lower with the GyneFix IUD (8.34) than with the TCu380A IUD (14.13) and results in a higher rate of continuation with the GyneFix IUD compared to the TCu380A IUD (90.73 vs 85.25). Neither perforations nor pelvic inflammatory disease cases were encountered with either device in this study, demonstrating the safety of the anchoring system. PMID- 10802274 TI - Formation of copper oxychloride and reactive oxygen species as causes of uterine injury during copper oxidation of Cu-IUD. AB - The lining of the uterus and cervix might be injured by a variety of oxidation products of Cu in a Cu-IUD, including cuprous ions, dissolved and precipitated cupric ions, and reactive oxygen species such as superoxide radicals, hydrogen peroxide, and hydroxyl radicals. In this study, the human amnious WISH cell line was employed as a model of uterine cells in the presence of copper. The cell viability was decreased by elemental copper, which was alleviated up to 70% by the addition of catalase. The addition of copper oxychloride caused cell death in a dose-dependent manner. Hydroxyl radicals in the presence of copper were determined by the formation of malondialdehyde. Soluble cuprous chloride complexes are formed in the uterus by slowly entering oxygen. The complexes are partly oxidized to insoluble copper oxychloride. which damages the endometrium. Unoxidized cuprous ions migrate to the oxygen-rich cervix and are oxidized to copper oxychloride, causing cervix damage. PMID- 10802275 TI - An open-label, multicenter study to evaluate Yasmin, a low-dose combination oral contraceptive containing drospirenone, a new progestogen. AB - This open-label, multicenter study evaluated the efficacy, safety, and cycle control of Yasmin, a new low-dose, monophasic oral contraceptive containing the unique progestogen drospirenone (DRSP) 3 mg and ethinyl estradiol (EE) 30 microg. DRSP is a synthetic progestogen that has antiandrogenic and antimineralocorticoid effects. In this study, 326 women were evaluated and 220 (67%) completed all 13 treatment cycles. The corrected Pearl Index was 0. 407. Of the 151 subjects who experienced intermenstrual bleeding at any time during the study, the majority (64%) had bleeding during only one or two pill cycles. Breakthrough bleeding without spotting occurred in 1% of all cycles, spotting without breakthrough bleeding in 9.3% of all cycles, and breakthrough bleeding with spotting in 3% of all cycles. Amenorrhea was observed in 3% of all cycles. In all, 20 subjects (6%) discontinued participation in the study because of adverse events. No serious adverse events related to the study drug were reported. No clinically significant changes in weight, blood pressure, or lipids were reported. The impact of the new progestogen DRSP on the women's self-perception of menstrual health was also evaluated. Subjects reported that symptoms of water retention, negative affect, and increased appetite significantly improved at cycle 6 from baseline. This study demonstrates that Yasmin is an effective oral contraceptive that is safe and well tolerated. PMID- 10802276 TI - Evaluating contraceptive choice through the method-mix approach. An Indian Council of Medical Research (ICMR) task force study. AB - The method-mix approach was used to evaluate informed contraceptive choices in the present study. A total of 8,077 potential clients were given a balanced presentation of all available contraceptive methods in the national program, ie, the CuT 200 intrauterine device (IUD), low-dose combined oral pills (OC), condom, and sterilization (female/male) along with a new method, Norplant(R).(1) The majority of women opted for spacing methods; among them, the IUD was preferred by about 60% of clients, followed by condoms (9%), OC (6%), and Norplant (5%). Sterilization, mainly female, was accepted by about 17% of the women making an informed choice. The economic status of couples did not influence the contraceptive choices, as all the methods were offered free of cost in the present study, which is the current practice in the national program. Illiterate women more often accepted sterilization (about 25%) than did literate women (15%). This is because illiterate women had more children; about 30% of illiterate women had three or more children, as opposed to 16.2% of literate women. However, literacy status did not influence the choice of any specific spacing method. The study also revealed that, by encouraging potential clients to make an informed choice, they could override the provider's bias while accepting a particular type of spacing method. This is evident from the observation that Norplant was the first choice of the provider for 35% of the women, whereas only 5% of women preferred and accepted Norplant. The present study stresses an urgent need to promote the practice of informed choices in the national program with a variety of contraceptive options-especially, spacing methods for improving contraceptive prevalence and reproductive health in the country. PMID- 10802277 TI - Characteristics of women associated with continuing instruction in the Creighton Model Fertility Care System. AB - Effective use of natural family planning is strongly dependent upon adequate instruction. The Creighton Model Fertility Care System (CrMS) has a standardized protocol for instruction of new users that includes individual follow-up visits in the first year of use. This study evaluated the number of follow-up visits completed by new CrMS users from eight CrMS centers in the United States. Four follow-up visits were completed by 75.7% of women. Women who continued instruction were more likely to be educated, Catholic, and white, and to have a more challenging reproductive status (discontinuing oral contraceptives, or breastfeeding). These results suggest that the number of follow-up visits needed varies among new CrMS users. Future research should address the optimal length of instruction for adequate use of the CrMS by women with different characteristics and needs. PMID- 10802279 TI - The effects of nomegestrol acetate subdermal implant (Uniplant) on serum cholesterol, triglycerides, and lipoproteins in Nigerian users. AB - The study was conducted to assess the effects of a nomegestrol acetate subdermal contraceptive implant (Uniplant(R)) on the lipid profiles of indigenous Nigerian women. Cholesterol content of the major lipoproteins, along with total cholesterol and triglycerides, were measured in fasting blood samples collected before implant insertion and then at months 1, 3, 6, and 12 of use. All volunteers were of the reproductive age, healthy, and had no contraindications to hormonal contraception. The mean levels of cholesterol and low density lipoprotein-cholesterol (LDL-C) increased gradually, albeit insignificantly, from insertion to month 12 of implant use. An initial decline in the mean levels of high density lipoprotein-cholesterol (HDL-C), reaching a minimal level (37.31 +/- 4.95 mg/dl) at month 3 of implant use, was observed. This was followed by a gradual rise that peaked (39.73 +/- 5.53 mg/dL) at month 12. These values were, however, not significantly different from the preinsertion value. The only significant difference (p <0. 005 ) was in the mean level of triglycerides at month 12 (90.90 +/- 17.75 mg/dL) when compared with the preinsertion mean level of 81.77 +/- 24.14 mg/dL. Both values were, however, within normal limits. These results indicate that Uniplant does not have a deleterious effect on the lipid profiles of Nigerian acceptors and can be safely prescribed to women seeking contraception. PMID- 10802278 TI - Vaginal epithelial surface appearances in women using vaginal rings for contraception. AB - Vaginal inspections using colposcopy before insertion of contraceptive vaginal rings and at 2-month intervals during ring use were conducted on 169 users of four different formulations. The rings studied released Nestorone alone (50, 75, 100 g daily over 6 months); ethinyl estradiol: Nestorone (30:100 and 15:150 g daily over 6 months); ethinylestradiol:norethindrone acetate (20:1000 and 15:1000 g daily over 4 months); and ethinyl estradiol:norethindrone acetate (20:1000 g daily over 12 months). A total of 88 altered or atypical conditions of the vaginal surface appearance were recorded in 507 inspections (17.4% of inspections). Many of these atypical appearances were quite subtle. The incidence was significantly higher (p <0.01) than in the single pretreatment examinations (11 in 158 inspections; 7.0%), but closely matched that of a "control group" of sexually active women who were the subject of an earlier study by the same investigators. In that study, the incidence was 18% (57 atypical conditions in 317 inspections). In all, 83% of atypical conditions identified in the vagina during ring use had disappeared by the next scheduled colposcopy despite continued ring use. Findings of potential significance were conservatively defined as all ulcerations, those abrasions and ecchymoses that were >0.5 cm in any direction, and fields of five or more petechiae. Findings fitting those criteria comprised 30% of atypical conditions in ring users, 33% in the control group, and 27% pretreatment. The corresponding incidence as a percentage of inspections were 5.3%, 6. 0%, and 2.5% in the ring users, control groups, and pretreatment groups, respectively. These differences were not statistically significant. The findings suggest that the vaginal rings included in the studies contributed little, if at all, to clinically significant lesions or to total lesion incidence. Further definition would require a larger and longer-term study with matched controls. PMID- 10802280 TI - Evaluation of chemiluminescent estradiol conjugates by using a surface plasmon resonance detector. AB - A series of chemiluminescent 17beta-estradiol probes were synthesized. Relative equilibrium dissociation constants (K(D)) for the interaction of an anti-E(2) Fab fragment for the probes in solution were evaluated using a single E(2)-analog biosensor surface on a BIAcore surface plasmon resonance instrument. The results show the antibody fragment binds all chemiluminescent conjugates tested with high affinity showing only minor preferences for site of substitution (C6 versus C7), stereochemistry (alpha versus beta), or linker moiety. PMID- 10802281 TI - Reactions of the four diastereomeric 16-amino-17-hydroxy-3-methoxy-estra 1,3,5(10)-trienes with aromatic ortho-hydroxy and heteroaromatic alpha-aldehydes and with 1, 3-dicarbonyl compounds-molecular structures of condensation products and of copper(II) complexes. AB - Vicinal amino alcohols of steroids have been used as starting materials for the synthesis of chiral ligands with defined arrangements of functional groups. Condensation of the four diastereomeric 16,17-steroid amino alcohols 1a-1d with aromatic o-hydroxy and heteroaromatic alpha-aldehydes afforded the Schiff bases 2 6. When the 16,17-substituted compounds 2d, 5d, 6a, and 6d were in solution, the isomeric oxazolidines were detectable by (1)H NMR spectroscopy. The formation of oxazolidines could be avoided by using bulky aldehydes. Reduction of the Schiff bases (also in mixtures with oxazolidines) with NaBH(4) yielded the new N substituted amino alcohols 12-15. The condensation products of 1a-1d with 1,3 dicarbonyl compounds (7 and 8) exhibited the 1-enamino-3-oxo structure ((1)H NMR spectroscopy). By means of X-ray analysis of 2a-2d, 3d, 7a, and 7c, the torsion angles for the 16N, 17O substituents, which are important for a participation of the 17O substituent in the complexation of metal ions, have been determined. Furthermore, a preferred arrangement between the chelate ring and the steroid plane existed in all investigated condensation products attributable to torsion angles 16H-C16-16N-C of 5-61 degrees. This arrangement was also preserved in the copper(II) complex 11 with 16alpha,17beta-trans configuration of the bidentate steroid ligand and a ratio of 2:1 for ligand: copper in contrast with dimeric copper(II) complexes with a tridentate steroid ligand of 16beta, 17beta-cis configuration (ratio of 1:1 for ligand:copper). The crystal structures of the condensation products are also discussed. In most cases, intermolecular hydrogen bonds between 17-hydroxy groups and the chelate oxygen caused polymeric strands. PMID- 10802282 TI - A mineralocorticoid-like receptor in the rainbow trout, Oncorhynchus mykiss: cloning and characterization of its steroid binding domain. AB - Using reverse transcriptase polymerase chain reaction (PCR) (RT-PCR) with degenerate primers followed by 3' rapid amplification of cDNA ends PCR (3'Race PCR) we have isolated a new fish steroid receptor cDNA sequence of 1806 bp from rainbow trout (Oncorhynchus mykiss) testis. This sequence has clear homology with various mineralocorticoid receptor cDNA sequences (rat, human, African toad: 68 70% amino acid identity), and encompasses the second part of DNA binding domain (C domain), the whole hinge region (D domain) and the steroid binding domain (E domain) plus 726 bp of 3'untranslated sequence. COS-1 cells transfected with a pCMV5 expression vector containing the whole E domain (pCMV5-rtMR) showed high affinity binding for cortisol (K(a) = 0.53+/-0.03 nM, K(d) = 1.9 nM) in the cytosol, which could not be detected in untransfected cells. Aldosterone displaced (3)H-cortisol binding, though was less effective by than unlabeled cortisol (P<0.05). Competition experiments with other steroids gave the following hierarchy for the displacement of the (3) dexamethasone, whereas 17, 20beta dihydroxy-4-pregnen-3-one and 17,20beta,21beta-trihydroxy-4 pregnen-3-one (two fish specific progestins) did not show any specific binding. These results strongly suggest that this cDNA sequence encodes a rainbow trout mineralocorticoid-like receptor, and represent the first description of such a receptor in teleost fish where aldosterone, the classic mineralocorticoid, is believed to be absent. PMID- 10802283 TI - Synthesis and bioactivity evaluation of brassinosteroid analogs. AB - Four new analogs of 28-homocastasterone have been synthesized and completely characterized for the first time from stigmasterol. (22R, 23R,24S)-3beta-acetoxy 22,23-dihydroxy-5alpha-stigmastan+ ++-6-one (17), (22R,23R,24S)-3beta-bromo-22,23 dihydroxy-5alpha-stigmast an-6-one (18), (22R,23R,24S)-3beta-acetoxy-5,22, 23 trihydroxy-5alpha-stigmastan-6-one (20), and (22R,23R, 24S)-3beta-bromo-5,22,23 trihydroxy-5alpha-stigmastan-6-one (21), were obtained through a synthetic route based on regioselective Delta(5) epoxidation. Compounds 17 and 18, bearing a 5alphaH moiety, were prepared through a reductive opening of the 5beta,6beta epoxy precursor, and compounds 20 and 21, analogs with a 5alphaOH moiety were obtained by hydrolytic opening of a mixture of 5alpha,6alpha and 5beta,6beta epoxy precursors. Known compounds 19 and 22 were also obtained following the described synthetic routes, respectively. The new compounds were tested with the traditional auxin-like bioassay for brassinosteroids with 19 and 22 as standards. All compounds were comparatively evaluated for their inhibitory effect on the replication of DNA (HSV-1) virus. PMID- 10802284 TI - Time-resolved fluoroimmunoassay of plasma daidzein and genistein. AB - We present a method for the determination of the phytoestrogens daidzein and genistein in plasma (serum). These weakly estrogenic isoflavones occur in soybeans and in smaller amounts in some other beans and plants. It has been suggested that they may afford protection against prostate and breast cancer. The method is based on time-resolved fluoroimmunoassay (TR-FIA) using a europium chelate as a label. After synthesis of 4'-O-carboxymethyl-daidzein and 4'-O carboxymethyl-genistein the compounds are coupled to bovine serum albumin (BSA), then used as antigens to immunize rabbits. The tracers with the europium chelate are synthesized using the same 4'-O-derivative of the isoflavones. After enzymatic hydrolysis and ether extraction the immunoassay is carried out using the VICTOR 1420 multilabel counter (Wallac Oy, Turku, Finland). The antisera cross-reacted to some extent with some isoflavonoids but not with flavonoids. The cross-reactivity seems not to influence the results, which were highly specific for both compounds. The correlation coefficients between the TR-FIA methods and the reference method based on isotope dilution gas chromatography-mass spectrometry were high; r-values were about 0.95-0.99 depending on concentration. The intra-assay coefficients of variation (CV%) for daidzein and genistein at three different concentrations vary 3.2-4.5 and 3.2-4.1, respectively. The inter assay CVs vary 5.0-6.3 and 4.5-5.3, respectively. The working ranges of the daidzein and genistein assays are 1.0-216 and 1.7-370 nmol/l, respectively. The plasma values (n = 80) of daidzein and genistein are very low in Finnish subjects (mean for daidzein, 3.8+/-6.8 and for genistein, 3.2+/-7.6 nmol/l; median value for daidzein 1.5 and for genistein 1.4 nmol/l). PMID- 10802285 TI - Synthesis and GABA(A) receptor activity of 6-oxa-analogs of neurosteroids. AB - The 6-oxasteroids 3alpha-hydroxy-6-oxa-5alpha-pregnan-20-one (3) and 3alpha hydroxy-6-oxa-5beta-pregnan-20-one (4) were obtained from pregnenolone acetate via the corresponding (5alpha or 5beta) 3beta, 20beta-diacetoxy-6-oxa-pregnane. Both steroids showed ca. 100-fold reduced potency for modulating [(3)H]flunitrazepam, [(3)H]muscimol or [(35)S]TBPS binding to the GABA(A) receptor when compared to their natural carbon analogs 3alpha-hydroxy-5alpha pregnan-20-one (1) and 3alpha-hydroxy-5beta-pregnan-20-one (2). PMID- 10802286 TI - Monoclonal antibodies against porcine macrophages. AB - Two mouse monoclonal antibodies (MAB) (clones 2G6 and 2B10) directed against porcine macrophages are described that are suitable for use in immunohistochemistry, FACS analysis and western blot. As immunogen, porcine cells from bronchoalveolar lavage (BAL) were used. The MABs obtained belonged to the mouse IgG1 subclass. The molecular weights of the corresponding antigens were detected by western blot under non-reducing conditions (2G6: 140-150kDa; 2B10: 140-145kDa). For specificity screening, porcine snap-frozen tissues of lung, lung lymph node, tonsil, spleen, thymus, brain, liver, gut and kidney were used. The MABs were able to identify cell populations of the mononuclear phagocytic system in these organs. While MAB 2G6 detected tissue macrophages (sinusoidal lymph node macrophages, red pulp spleen macrophages, Kupffer cells, Langerhans cells, thymus macrophages, macrophages of lung and macrophages of kidney), MAB 2B10 stained cells scattered in the lymph node (subsinusoidal, interfollicular and follicular macrophages) and in the lung interstitium. Additionally, it showed reactivity with Kupffer cells, spleen and kidney macrophages. An immunoreactivity of the MABs could be established also for human but not for bovine and avian macrophages. By flow cytometric analysis, MAB 2B10 reacted with a subpopulation of BAL and peritoneal cells. Antibody 2G6 detected macrophages of the BAL and the peritoneal fluid as well as peripheral blood monocytes. PMID- 10802287 TI - Assessing humoral and cell-mediated immune response in Hawaiian green turtles, Chelonia mydas. AB - Seven immature green turtles, Chelonia mydas, captured from Kaneohe Bay on the island of Oahu were used to evaluate methods for assessing their immune response. Two turtles each were immunized intramuscularly with egg white lysozyme (EWL) in Freund's complete adjuvant, Gerbu, or ISA-70; a seventh turtle was immunized with saline only and served as a control. Humoral immune response was measured with an indirect enzyme linked immunosorbent assay (ELISA). Cell-mediated immune response was measured using in vitro cell proliferation assays (CPA) using whole blood or peripheral blood mononuclear cells (PBM) cultured with concanavalin A (ConA), phytohaemagglutinin (PHA), or soluble egg EWL antigen. All turtles, except for one immunized with Gerbu and the control, produced a detectable humoral immune response by 6 weeks which persisted for at least 14 weeks after a single immunization. All turtles produced an anamnestic humoral immune response after secondary immunization. Antigen specific cell-mediated immune response in PBM was seen in all turtles either after primary or secondary immunization, but it was not as consistent as humoral immune response; antigen specific cell-mediated immune response in whole blood was rarely seen. Mononuclear cells had significantly higher stimulation indices than whole blood regardless of adjuvant, however, results with whole blood had lower variability. Both Gerbu and ISA-70 appeared to potentiate the cell-mediated immune response when PBM or whole blood were cultured with PHA. This is the first time cell proliferation assays have been compared between whole blood and PBM for reptiles. This is also the first demonstration of antigen specific cell-mediated response in reptiles. Cell proliferation assays allowed us to evaluate the cell-mediated immune response of green turtles. However, CPA may be less reliable than ELISA for detecting antigen specific immune response. Either of the three adjuvants appears suitable to safely elicit a detectable immune response in green turtles. PMID- 10802288 TI - Cellular immune response of lymph nodes from dogs following the intradermal injection of a recombinant antigen corresponding to a 66 kDa protein of Echinococcus granulosus. AB - A recombinant polypeptide (referred to as EgA31), which represents a 66kDa protein, was prepared from an Echinococcus granulosus cDNA library. In order to assess its potential to induce cellular immune responses, dog popliteal and prescapular lymph nodes were sensitized with this recombinant polypeptide. Subpopulations of lymphocytes were then analyzed by flow cytometry and immunohistochemistry on lymph node sections. Five days after the sensitization, the paracortical areas of the lymph nodes appeared hypertrophic, the number of CD3+, CD4+, CD8+ and CD5+ cells increased, the number of B-cells began to augment and some secondary follicles occurred, and a number of CD4+ cells appeared in germinal centers. Many large secondary follicles and a significantly augmented number of CD5+ cells in cords of medullae were observed 10 days after the sensitization. These active cellular responses strengthen the interest for further studies on the development of a vaccine with this recombinant polypeptide. PMID- 10802289 TI - Immune responses of chickens to dietary protein antigens. I. Induction of systemic and intestinal immune responses following oral administration of soluble proteins in the absence of adjuvant. AB - Oral administration of protein antigens in solution leads to the development of oral tolerance in most mammals but rarely so in the chicken. As dietary proteins are not expected to be immunogenic, the present study was undertaken to evaluate immunological consequences following oral exposure to protein antigens in chicks, and to determine whether or not this form of antigen is ignored. Chicks and turkey poults were fed solutions containing bovine serum albumin (BSA), porcine serum albumin, beta-lactoglobulin or bovine hemoglobin over a period of 6 days (25mg/chick/day). At different time points after feeding serum and bile were examined for presence of specific antibodies by ELISA. Surprisingly, the fed antigens induced robust antibody responses in the absence of added adjuvant. This immune response was further characterised to show that (1) a daily feeding regimen was more immunogenic than single dose feedings, (2) by using a daily feeding regimen, as little as 2mg/chick/day was fully immunogenic, (3) effective immunization was attained in chicks older than 10 day of age, (4) the main antibody class in the serum was IgG, and (5) high IgA levels were detected in the bile after booster feedings. These observations are difficult to reconcile with current concepts on peripheral tolerance to innocuous antigens, and indicate that the bird regulates tolerance and response in a manner different from that described in mammals. PMID- 10802290 TI - Molecular cloning of a carp Jak3 from activated leukocytes. AB - Janus kinases (Jaks) are involved in a signalling pathway leading to tyrosine phosphorylation, which is the key step in transducing cytokine signals from the external environment to the nucleus. We report here the molecular cloning of carp Jak3. A partial cDNA of the Jak homologue was initially identified from a cDNA pool obtained by subtracting the cDNAs from Con A-induced carp head kidney leukocytes with those from uninduced carp head kidney leukocytes. The entire coding sequence was assembled by sequencing both strands of cDNA clones amplified by using anchored PCR reaction. The complete cDNA was cloned by specific primers designed against the partial cDNAs and then sequenced to assure accuracy of the complete sequence. Sequence analysis reveals an ORF encoding a protein of 1026 amino acids, with the unique presence of two carboxy-terminal kinase and pseudokinase domains. The cDNA clone shows approximately 52-50%, 50-48.2%, 42-39% and 35-39% amino acid identity with the known Jak3s, Jak2s, Jak1s and Tyk2s, respectively. The sequence of the amino acid residues within the N-terminal JH domains of the carp Jak homologue shows higher similarities with the regions of human Jak3 than with human Jak2. The carp Jak is expressed as a single 4. 5Kb transcript at low levels in the head kidney and the spleen, with the RNA level increasing when leukocytes are activated with con A. The trace transcript was also detected by RT-PCR in the carp liver, muscle and skin, but not in the egg sample. Sequence analysis and pattern of expression suggest that this cDNA represents the carp Jak3 (accession number in GenBank: AF148993). PMID- 10802291 TI - The influence of ketone bodies and glucose on interferon, tumor necrosis factor production and NO release in bovine aorta endothelial cells. AB - Bovine aorta endothelial cells (BAECs) were used to determine the effect of ketone bodies and glucose on in vitro interferon (IFN), tumor necrosis factor (TNF) and nitric oxide (NO) production. BAECs were incubated for 4 and 24h with the ketone bodies: 3.8mmol/l beta-hydroxybutyrate (BHB), 1mmol/l acetoacetate (AcAc) and 5. 2mmol/l acetone (Ac), used separately or in a mixture together with cytokine inducers: Newcastle disease virus (NDV) and lipopolysaccharide (LPS). BHB alone (but not AcAc or Ac) and a mixture of ketone bodies caused a significant decrease in IFN titers induced by NDV and LPS and in TNF titers induced by LPS. Glucose used at concentrations of 5.55, 3.33 and 1.66mmol/l did not influence cytokine production.NO measured by the nitrite content in culture medium was released spontaneously from BAECs. A slight enhancement of NO release was observed after infection of BAECs with NDV; however, treatment with LPS caused inhibition of the release. The mixture of ketone bodies used with NDV or LPS enhanced NO release. However, when cells were incubated in the medium with 1. 66mmol/l glucose (mimicking low plasma glucose level in ketotic cows) a significant decrease in NO release was observed. This enhancing effect of ketone bodies and inhibition by low glucose in the final effect balanced each other, and the amounts of NO released in the medium with 1.66mmol/l glucose and with the mixture of ketone bodies resembled those produced at 3.33mmol/l glucose without ketone bodies. The significance of these effects of ketone bodies and glucose concentrations on cytokine and NO production in the immunity of ketotic cows has been discussed. PMID- 10802292 TI - In vivo T-cell subset depletion suggests that CD4+ T-cells and a humoral immune response are important for the elimination of orf virus from the skin of sheep. AB - In vivo lymphocyte subset depletion offers a unique opportunity to study the roles of different cellular components of the immune system of sheep during infection with orf virus. Lambs were depleted of specific lymphocyte subsets by the intravenous administration of monoclonal antibodies against ovine lymphocyte surface markers and then challenged with orf virus. The skin lesions that developed were scored visually as to their severity. Blood samples were collected to monitor the lymphocyte depletions and to measure orf-virus-specific antibody levels. Skin biopsies were collected from the lesion site and studied to determine the course of the infection and the presence of various cell types and orf virus. All the sheep developed orf virus lesions after infection. All three of the CD4-depleted lambs were unable to clear virus from their skin and did not have an antibody response to the virus. Virus was also detected in the skin of one each of the three CD8-depleted, WC1-depleted and control sheep on the final day of the trial. CD8(+) lymphocytes did not appear to be essential for viral clearance later in the infection. Depletion of the majority of gammadelta(+) T cells did not affect the outcome of orf virus infection. In sheep with high orf virus-specific antibody titres at the time of infection, orf lesions healed faster than lesions in sheep with low antibody levels, and this occurred regardless of the lymphocyte depletion status of the animals. This study suggests that the presence of CD4(+) T-cells and orf-virus-specific antibodies are important for the control of viral replication in the skin of infected sheep. PMID- 10802293 TI - IgE-reactivity to major Japanese cedar (Cryptomeria japonica) pollen allergens (Cry j 1 and Cry j 2) by ELISA in dogs with atopic dermatitis. AB - The present study investigated IgE-reactivity to two major Japanese cedar (Cryptomeria japonica, C. japonica) pollen allergens (Cry j 1 and Cry j 2) in dogs with atopic dermatitis by use of a fluorometric ELISA. The serum samples from 27 dogs that showed IgE-sensitivity to crude C. japonica pollen allergen by ELISA were tested for specific IgE to the two major allergens. All 27 dogs had anti-Cry j 1 IgE, and 10 (37%) had anti-Cry j 2 IgE. Inhibition of binding of dog specific IgE to crude C. japonica pollen allergen was carried out by addition of Cry j 1. When serum samples containing anti-Cry j 1 IgE but no anti-Cry j 2 IgE were incubated with Cry j 1, specific IgE binding to crude C. japonica pollen allergen was almost abolished. These findings suggest that Cry j 1 is a major allergen in dogs. PMID- 10802294 TI - Murine model study of the practical implication of trypanosome-induced immunosuppression in vaccine-based disease control programmes. AB - The relevance of trypanosome-induced immunosuppression in relation to the efficacy of vaccine-induced immunity was studied in mice. Mice were immunised with crude Trichinella spiralis muscle larvae homogenate vaccine and infected with T. spiralis and/or Trypanosoma brucei. Vaccination significantly decreased adult worm burden (p<0. 05) and accelerated worm expulsion in mice infected with T. spiralis only. T. brucei superinfection resulted in monocytosis, suppressed eosinophilia, significant decrease in PCV (p<0.001), higher numbers of adult worms (p<0.001) and failure to expel all adult worms by Day 12 post infection (p.i.). Regardless, they produced anti-Trichinella IgG(1) responses similar to those of the vaccinated non-T. brucei-infected group. T. brucei also suppressed the proliferative responses of spleen cells to stimulation with Con A and T. spiralis antigen, and induced strong production of interferon-gamma (IFN-gamma) in culture supernatants of antigen stimulated spleen and mesenteric lymph node cells. Interleukin-5 (IL-5) production was suppressed by T. brucei in supernatants of Con A- and antigen-stimulated spleen cells. It was concluded that trypanosome infections and the associated immunosuppression are of great practical significance in trypanosome endemic areas, especially with regards to disease control programmes involving vaccine-induced herd immunity. PMID- 10802295 TI - Cloning and characterization of bovine low molecular weight GTPases (Rac1 and Rac2) and rho GDP-dissociation inhibitor 2 (D4-GDI). AB - GTPases of the Rho family play important roles in human leukocyte signal transduction pathways; however, little is known about the function of these proteins in bovine cells. In the present studies, we isolated molecular clones of bovine Rac1, Rac2, and the Rac/Rho GTPase regulatory protein D4-GDP dissociation inhibitor (D4-GDI) from a bovine bone marrow cDNA library. These clones contained complete open reading frames, encoding 192, 192, and 200 amino acids, respectively. Comparison of the bovine amino acid sequences with those of other species demonstrated a high degree of identity of these proteins across all species, suggesting that these proteins likely play conserved functional roles in bovine leukocyte signal transduction pathways. Comparative Western blotting of these proteins in human and bovine neutrophil cytosol demonstrated that Rac2 was the predominant Rac species and that D4-GDI was the predominant GDI species in bovine neutrophil cytosol. Despite the high degree of homology between human and bovine Rac2, some of the anti-peptide antibody probes prepared against human Rac2 failed to recognize the bovine homologue. We also showed by subcellular fractionation techniques that Rac2 is localized primarily to the cytosolic compartment of resting bovine neutrophils, but is translocated to the plasma membrane after stimulation with PMA. These findings suggest that Rac2 does play a role in bovine neutrophil activation. In addition, these data will be helpful in developing more specific probes for investigating the role of these proteins in bovine leukocyte signal transduction pathways and for studying various inflammatory diseases in cattle. PMID- 10802296 TI - Inhibitory effect of stromal cell derived factor-1 on the replication of divergent strains of feline immunodeficiency virus in a feline T-lymphoid cell line. AB - The effect of a CXC-chemokine, stromal cell derived factor-1 (SDF-1), on the replication of divergent strains of feline immunodeficiency virus (FIV) was examined in order to identify the mechanism of cell entry of FIV. A chemotaxis assay, using a modified Boyden chamber method, confirmed the biological activity of recombinant human (rh) SDF-1 for a feline T-lymphoid cell line (Kumi-1). The viral replication of FIV, as measured by the reverse transcriptase (RT) activity in the culture supernatant, was significantly suppressed by addition of rhSDF-1 in a dose-dependent manner in Kumi-1 cells. Furthermore, PCR analysis of the FIV proviral genome indicated that the inhibitory effect of rhSDF-1 on the replication of FIV in Kumi-1 cells was due to the inhibitory effect in the early event of replication. The inhibitory effect on viral replication by exogenous rhSDF-1 was shown for four divergent FIV isolates of subtypes A, B, and D in Kumi 1 cells. PMID- 10802298 TI - Preface PMID- 10802297 TI - Dietary lutein stimulates immune response in the canine. AB - The possible immuno-modulatory action of dietary lutein in dogs is not known. Female Beagle dogs (17-18-month old; 11.4+/-0.4kg body weight) were supplemented daily with 0, 5, 10 or 20mg lutein for 12 weeks. Delayed-type hypersensitivity (DTH) response to saline, phytohemagglutinin (PHA) and a polyvalent vaccine was assessed on Weeks 0, 6 and 12. Blood was sampled on Weeks 0, 2, 4, 8 and 12 to assess (1) lymphocyte proliferative response to PHA, concanavalin A (Con A), and pokeweed mitogen (PWM), (2) changes in peripheral blood mononuclear cell (PBMC) populations, (3) interleukin-2 (IL-2) production and (4) IgG and IgM production. After the completion of 12-week study, we continued to collect the blood weekly up to 17 weeks to evaluate the changes in immunoglobulin production upon first and second antigenic challenges on Weeks 13 and 15. Plasma lutein+zeaxanthin was undetectable in unsupplemented dogs but concentrations increased (P<0.05) rapidly on Week 2 in lutein-supplemented dogs. Thereafter, concentrations generally continued to increase in dose-dependent manner, albeit at a much slower rate. Dogs fed lutein had heightened DTH response to PHA and vaccine by Week 6. Dietary lutein increased (P<0.05) lymphocyte proliferative response to all three mitogens and increased the percentages of cells expressing CD5, CD4, CD8 and major histocompatibility complex class II (MHC II) molecules. The production of IgG increased (P<0.05) in lutein-fed dogs after the second antigenic challenge. Lutein did not influence the expression of CD21 lymphocyte marker, plasma IgM or IL-2 production. Therefore, dietary lutein stimulated both cell-mediated and humoral immune responses in the domestic canine. PMID- 10802299 TI - Intracerebellar conditioning--Brogden and Gantt revisited. AB - In 1942, Brogden and Gantt reported that electrical stimulation of cerebellar white matter elicited specific behavioral responses (limb flexion, eyeblink, etc.) and that these movements so elicited could easily be conditioned to a neural tone CS, using standard Pavlovian procedures. This early evidence for the key role of the cerebellum in learning of discrete movements has in recent years been replicated and much extended. It is now clear that the cerebellum is the essential structure for associative learning of discrete movements elicited by peripheral aversive or intracerebellar stimuli and that the memory traces so formed are stored in the cerebellum. PMID- 10802300 TI - Brain substrates of classical eyeblink conditioning: a highly localized but also distributed system. AB - The rabbit classical nictitating membrane/eyeblink conditioning preparation has proven highly valuable for delineating neural structures and systems involved in associative learning. Research conducted over the last 20 years has revealed that the essential neural circuitry for acquisition and performance of this simple, learned, motor response resides in the cerebellum and related brain stem structures. While this system appears to be highly localized, many other brain areas are recruited during eyeblink conditioning. Further, involvement of the cerebellum in associative learning and memory seems to be limited by certain parametric conditions present at the time of learning. These data suggest that classical eyeblink conditioning can also be characterized as a distributed system. Data in support of the highly localized, yet distributed nature of the neural systems involved in classical eyeblink conditioning are presented and discussed here. PMID- 10802301 TI - Multiple memory systems, development and conditioning. AB - A century of behavioral and neurobiological research suggests that Pavlovian conditioning involves three component memory systems: sensorimotor, affective and cognitive. In classical eyeblink conditioning, there is evidence that these three memory systems involve, respectively, the cerebellum, amygdala and hippocampus. This article reviews developmental research on eyeblink conditioning in rodents that is beginning to characterize ontogenetic dissociations and interactions among these memory systems. This research shows that the functional development of the affective system (conditioned fear response) precedes that of the sensorimotor system (conditioned eyeblink reflex). Modulation of these two systems by cognitive processes also seems to emerge at different points in ontogeny. Implications for cognitive development and research on multiple memory systems are discussed. PMID- 10802302 TI - The modulation of Pavlovian memory. AB - Exposure to stressful experiences as well as sex differences in the brain are known to influence the acquisition of new memories. This review focuses on acquisition of two types of Pavlovian learning paradigms: hippocampal-independent delay conditioning and hippocampal-dependent trace conditioning and their modulation by exposure to stressful experience and sex differences in the brain. We concentrate on two sets of findings: the first is that exposure to an acute stressful experience enhances Pavlovian conditioning in the male rat, while exposure to the very same experience dramatically impairs conditioning in female rat. The sexually-opposed effects of stress on conditioning are mediated by differing hormonal substrates (adrenal versus ovarian steroids) and possibly by differing anatomical and biochemical pathways. The second set of findings is that training with hippocampal-dependent trace conditioning enhances the survival of newly generated neurons in the adult hippocampal formation. The same amount of training with hippocampal-independent delay conditioning does not affect their survival. In addition, females acquire the trace task faster than males and generate more new neurons. As with the stress effects on learning, these sex effects are influenced by hormonal status. It is our contention that identifying the hormonal and neuronal processes that modulate associative memory formation will provide insight into the processes of memory formation itself. PMID- 10802303 TI - The tractable contribution of synapses and their component molecules to individual differences in learning. AB - Though once of central importance to psychologists and neurophysiologists alike, the elucidation of neural substrates for individual differences in learning no longer attracts a broad research effort and occupies a place of largely historical interest to the contemporary disciplines. The decline in interest in this subject ensued in part from the perception, arrived at decades ago, that individual differences in learning were not quantified as easily as had once been presumed. Furthermore, the dominant hypotheses in the field defied testing within the constraints imposed by the complex and largely inaccessible vertebrate nervous system. Using a 'model systems' approach where the individual cells and synaptic interactions that comprise a neural network can be identified, we have returned to this question and have established a framework by which we can begin to discern the basis for much of the variability between individuals in their capacity to learn. In the marine mollusc Hermissenda, we have found that a common influence on transmitter exocytosis is expressed homogeneously throughout the nervous system regardless of transmitter system or receptor class. Though uniformly expressed within an individual, this influence on synaptic efficacy is differentially expressed between animals. Importantly, the basal efficiency of exocytosis expressed in an individual nervous system is strongly correlated with the degree to which activity-dependent forms of neuronal/synaptic facilitation can be induced in that nervous system, and predicts the capacity for the intact animal to learn a Pavlovian association. Furthermore, we have established that a decline in basal synaptic efficacy in aged animals, arising from chronic presynaptic Ca(2+) 'leak', may contribute to age-related learning impairments. Because certain fundamental components of the exocytotic cascade are conserved widely across cell types, transmitter systems and species, the principles that we describe may have broad implications for understanding normal variability in learning, but also, in the development of specific strategies to compensate for mild learning deficits and age-related cognitive decline. PMID- 10802304 TI - A componential view of configural cues in generalization and discrimination in Pavlovian conditioning. AB - This paper describes three theoretical approaches to the representation of configural cues in generalization and discrimination in Pavlovian conditioning: that of the Rescorla-Wagner model, the Pearce model, and the authors' 'replaced elements' model. We summarize the results of a generalization experiment using the rabbit Pavlovian conditioned eyeblink response where animals were trained with cues A, AB, or ABC, and tested with A, AB, and ABC. The pattern of generalization decrement in testing supported the replaced elements model. PMID- 10802305 TI - Contextual fear, gestalt memories, and the hippocampus. AB - This review examines the relationship between exploration and contextual fear conditioning. The fear acquired to places or contexts associated with aversive events is a form of Pavlovian conditioning. However, an initial period of exploration is necessary to allow the animal to form an integrated memory of the features of the context before conditioning can take place. The hippocampal formation plays a critical role in this process. Cells within the dorsal hippocampus are involved in the formation, storage and consolidation of this integrated representation of context. Projections from the subiculum to the nucleus accumbens regulate the exploration necessary for the acquisition of information about the features of the context. This model explains why electrolytic but not excitotoxic lesions of the dorsal hippocampus cause enhanced exploratory activity but both cause deficits in contextual fear. It also explains why retrograde amnesia of contextual fear is greater than anterograde amnesia. PMID- 10802306 TI - Is the hippocampus necessary for contextual fear conditioning? AB - The hippocampus is widely believed to be essential for learning about the context in which conditioning occurs. This view is based primarily on evidence that lesions of the dorsal hippocampus disrupt freezing to contextual cues after fear conditioning. However, lesions that disrupt freezing produce no effect on fear potentiated startle, a second measure of contextual fear. Moreover, hippocampal lesions also do not disrupt the contextual 'blocking' phenomenon, which provides an indirect measure of contextual fear. In these paradigms, at least, it appears that hippocampal lesions disrupt the expression of freezing, rather than contextual fear itself. This interpretation is supported by the finding that rats showing preserved contextual blocking after hippocampal lesions show deficits not only in contextual freezing, but also in unconditioned freezing. These findings are consistent with a growing body of data from other conditioning paradigms that contextual learning is spared after lesions of the dorsal hippocampus. Nonetheless, there remain some reports of impaired contextual fear conditioning after hippocampal lesions that cannot be attributed easily to a disruption of freezing. Thus, it is concluded that the hippocampus may be involved in contextual learning under certain--as yet, unspecified--circumstances, but is not critical for contextual learning in general. PMID- 10802307 TI - The hippocampus and contextual memory retrieval in Pavlovian conditioning. AB - Several theories of hippocampal function have suggested a role for the hippocampus in contextual memory retrieval. However, these theories are based on studies using permanent pretraining lesions, which confound the role of the hippocampus in learning with its role in retrieval. We argue that an alternative methodology is required to assess the role of the hippocampus in memory retrieval processes. This alternative methodology involves temporary inactivation of the hippocampus in Pavlovian paradigms that lend themselves to an examination of retrieval. An example of this approach is considered in a Pavlovian fear conditioning paradigm. We examined the influence of hippocampal inactivation on the context-specific expression of latent inhibition, a response decrement displayed when excitatory conditioning is preceded by non-reinforced presentations of a to-be-conditioned stimulus. Reversible inactivation of the dorsal hippocampus prior to retrieval testing eliminated the context-specific expression of latent inhibition. The nature of the hippocampal role in retrieval processes is discussed in the light of these data. PMID- 10802308 TI - Shock sensitization of startle: learned or unlearned fear? AB - Following shock, rats exhibit a potentiated startle response to a sudden, loud noise. It has been suggested that this shock sensitization of the startle response can be used as a model preparation for studying unlearned fear. After reviewing the theoretical and empirical bases for this claim, the results of several recent studies that show that the shock sensitization of startle effect is actually mediated by contextually conditioned fear are presented. From this, it is concluded that the shock sensitization of startle procedure is an appropriate model preparation for studying contextual conditioning but is not an appropriate procedure for studying unlearned fear. Several other procedures that have potential as model preparations for studying unlearned fear are discussed briefly. PMID- 10802309 TI - Behavioral and neuropsychological foundations of olfactory fear conditioning. AB - Pavlovian fear conditioning procedures have been a fruitful means of exploring the neural substrates of associative learning. There is now substantial evidence suggesting that many aspects of conditioned fear depend critically upon the integrity of the amygdala and the perirhinal cortex. Recent studies in our laboratory examining the contributions of these areas to olfactory and contextual fear conditioning are reviewed; collectively the results of these studies suggest that the amygdala participates critically in the acquisition and expression of fear conditioned to both an olfactory conditioned stimulus (CS) and to the training context, while the perirhinal cortex contributes to olfactory, but not contextual, fear conditioning. Moreover, it appears that perirhinal cortex may play a prominent role in recognition of the CS following conditioning. These results are discussed in light of the extent to which they replicate and extend previous research examining the contributions of these areas to fear conditioned to auditory and visual CSs. PMID- 10802310 TI - Pavlovian conditioning of immune function: animal investigation and the challenge of human application. AB - Pavlovian conditioning of immune functions provided early impetus to the rapidly expanding knowledge of bi-directional communication among the immune, endocrine, and central nervous systems. Since these early investigations, the phenomenology of this response has been well characterized. However the neural mechanisms and biological relevance of conditioned immunomodulation remain unclear. To this end, we present here data from our laboratories that have: (1) revealed some of the neural mechanisms and biological relevance of an animal model of conditioned immunomodulation; (2) demonstrated the conditionability and potential mechanisms of conditioned immune responses in healthy humans, and (3) investigated conditioned immunomodulation in a clinical sample. Together, these data demonstrate that animal models provide a basis for investigating mechanisms whereby conditioned changes in immune function may modulate health status in a clinical realm. PMID- 10802311 TI - Classically conditioned responses following repeated insulin and glucose administration in humans. AB - This paper describes the neural basis and the role of Pavlovian conditioning in the modification of blood glucose and related endocrine parameters after repeated insulin and glucose administration. Pavlovian conditioning requires that conditioned stimulus (CS) and unconditioned stimulus (US) are both detected in the central nervous system (CNS), where the CS-US association takes place. We will therefore elucidate the detectability of insulin and glucose in the CNS. Since current data focus almost exclusively on animals, we conducted a placebo controlled insulin conditioning experiment in humans (Experiment 1). Compared with the control group with CS-placebo pairings throughout, the experimental group with previous CS-insulin pairings in the acquisition phase showed a conditioned decrease in blood glucose and a trend for a conditioned baseline insulin increase, and an increase in cortisol levels relative to baseline and cumulative number of neuroglycopenic symptoms in the CS-placebo test session. The conditionability of glucose administration also had to be examined; experiments using an arbitrary CS and glucose are extremely rare, even in animals. Glucose is the natural stimulus for endogenous insulin secretion, so studies on cephalic phase insulin release (CPIR) will be reviewed in this paper. We implemented a placebo-controlled three-group design (Experiment 2): Subjects received either CS insulin, CS-glucose or CS-placebo pairings during the acquisition. Together, our results demonstrate the conditionability mainly of insulin, but also of glucose effects in healthy humans. The clinical relevance and future research perspectives are outlined with an emphasis on insulin in the brain and its role in learning and memory. PMID- 10802312 TI - Anxiety and helplessness in the face of stress predisposes, precipitates, and sustains gastric ulceration. AB - When an organism is subjected to stress, gastric ulcers or ulcerations commonly develop but the vulnerability to and amount of pathology induced varies considerably between individuals. The role of psychological factors in determining the occurrence and severity of these ulcerations is amply demonstrated in the studies reviewed here. The present paper (a) gives a brief history of the search for the causes of gastric ulcer, (b) provides a review of our own research which reveals that vulnerability to gastric ulceration is modulated by psychologically meaningful experiences, and (c) offers a multifactorial perspective on the causes of gastric ulceration and the future of research on it. PMID- 10802313 TI - Pavlovian influences over food and drug intake. AB - Consuming food and taking drugs share several important characteristics. In particular, each causes changes in important physiological parameters that are constantly being monitored and regulated by the brain. As examples, blood glucose increases after meals; and body temperature decreases after ethanol is taken. Such changes elicit neurally-mediated homeostatic responses that serve to reduce the magnitude and duration of the perturbation. It is argued that when an individual can accurately anticipate pending meals or drugs, it can make appropriate responses to minimize or totally neutralize the meal/drug-elicited perturbations. This phenomenon, which is the basis for meal and drug tolerance, relies upon Pavlovian conditioning. Literature is reviewed which documents the role of conditioning processes in the development of tolerance. The argument is made that conditioned responses enable individuals to derive necessary or desirable aspects of food and drugs while minimizing some of their negative effects. In a final section, drug tolerance is discussed as a natural consequence of evolution-derived, meal-related learning processes, with associated negative consequences. PMID- 10802314 TI - Characterization of cytochrome b from Toxoplasma gondii and Q(o) domain mutations as a mechanism of atovaquone-resistance. AB - Atovaquone is active in vitro against the tachyzoites of Toxoplasma gondii at nanomolar concentrations and is used clinically to treat acute cases of human toxoplasmosis. In pursuit of the mechanism of action of atovaquone against T. gondii and to understand how resistance might arise, drug-resistant mutants were generated and examined. The previously uncloned cytochrome b gene of T. gondii was cloned and sequenced from wild type and resistant strains as this was a likely candidate for the target of the drug and thus a source of resistance. Mutations are present within the cytochrome b gene of atovaquone-resistant parasites (M129L and I254L) and represent alterations in two different regions of the ubiquinol-binding pocket (Q(o) domain) of cytochrome b, suggesting that atovaquone interferes with electron transport at the cytochrome bc(1) complex in T. gondii. A structural model for how this hydroxynaphthoquinone is binding within the Q(o) domain is presented. Further analysis of the cytochrome b gene suggested that the protein may differ from other homologues by terminating within the mitochondrial membrane. Cytochrome b becomes the first complete mitochondrial gene and cognate protein to be described for T. gondii. PMID- 10802315 TI - The tyrosine-86 allele of the pfmdr1 gene of Plasmodium falciparum is associated with increased sensitivity to the anti-malarials mefloquine and artemisinin. AB - Although chloroquine-resistance (CQR) in Plasmodium falciparum is increasing and resistance to other blood schizonticidal anti-malarials has been reported, the molecular basis remains unclear. In this study fresh field isolates were obtained from The Gambia, an area of emerging CQR and tested for sensitivity to the anti malarial drugs mefloquine, halofantrine, artemisinin, dihydroartemisinin, chloroquine and quinine. Sequence polymorphisms in the pfmdr1 gene and size polymorphisms in the cg2 gene were assessed using PCR-based systems. A strong association was observed between the presence of the tyr-86 allele of pfmdr1 and increased sensitivity to mefloquine and halofantrine, as well as the structurally unrelated drugs artemisinin and dihydroartemisinin. A weaker association was found between the presence of tyr-86 and increased resistance to chloroquine and quinine. The cg2 Dd2-like omega repeat size polymorphism was associated with increased resistance to chloroquine and increased sensitivity to mefloquine and halofantrine. An intragenic association was also found between a polymorphism in the polyasparagine linker region of pfmdr1 and the tyr-86 allele, which may be due to genetic hitchhiking, indicative of recent selection by chloroquine. Our data support a hypothesis where the pfmdr1 gene confers a true multidrug resistance phenotype which is lost by mutation. PMID- 10802316 TI - Characterisation of the chondroitin sulphate of Saimiri brain microvascular endothelial cells involved in Plasmodium falciparum cytoadhesion. AB - Cytoadhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to chondroitin 4-sulphate (CSA) is inhibited by soluble CSA in vitro on Saimiri brain microvascular endothelial cells (SBEC) and in vivo in P. falciparum-infected Saimiri monkeys. We tested whether the SBEC model was appropriate for studying CSA-binding IRBC using four cell lines. All SBEC expressed a chondroitin sulphate (CS), with a composition of CSA. The mean sizes of these CSA were 20.5, 22, 23, 32.5 and 36 kDa for SBEC 3A and C2, CHO, SBEC 1D and 17, respectively. We found that cytoadhesion of the Palo-Alto (FUP)1 CSA-binding phenotype, selected by panning on SBEC 17, was specifically inhibited in a dose-dependent manner by all the purified CSA. The extent of inhibition depended on the cellular origin of the tested CSA. SBEC 17 CSA was 33 times more efficient than CHO-CSA and 21 times more efficient than the 50 kDa commercial bovine trachaea CSA. Dynabeads coated with a total extract of SBEC 1D CS-proteoglycans interacted with CSA- but not with CD36- or ICAM-1-binding IRBC. These Dynabeads also interacted specifically with the PfEMP1 DBL-3 domain, on the surface of CHO transfectants, but not with the CIDR-1 domain. Thrombomodulin was involved in IRBC adhesion to all SBEC whereas CD44 was only expressed by SBEC 1D and 17. These two CSA-proteoglycans have also been detected at the surface of human endothelial cells. Thus, the two homologous models, SBEC/Saimiri sciureus, are useful and reliable tools for the evaluation of new anti-CSA adhesion treatments and anti-disease vaccines for pregnant women. PMID- 10802317 TI - Molecular characterization of a novel microneme antigen in Neospora caninum. AB - The apical complex of the parasites belonging to the phylum Sporozoa is believed to be critically involved in the events leading to host cell invasion. The characterization of the components of this subcellular structure is therefore an important step towards understanding how these parasites achieve host cell entry. Affinity-purification of an anti-Neospora caninum antiserum on a reactive protein band of approximately 40 kDa following Triton-X-114 extraction of parasite proteins, SDS-PAGE and Western blotting, yielded an immunoglobulin fraction which, by immunofluorescence, stained predominantly the apical portion of N. caninum tachyzoites. Following immunoscreening of a N. caninum tachyzoite lambdagt22 cDNA expression library, the respective full length cDNA sequence was determined. This sequence was found to encode a protein of 362 amino acids, with a calculated Mr of 38086. This protein is encoded by a single copy gene which produces a transcript of 2.4 kb. Sequence analysis showed that it contains a N terminal putative signal peptide sequence and two potential membrane spanning regions. Four consecutive epidermal growth factor like domains were identified, as well a conserved sequence motif for binding of ATP/GTP (P-loop). The full length cDNA was expressed as a recombinant poly-histidine fusion protein in Escherichia coli, and antibodies affinity purified on this protein labelled exclusively a 38 kDa band on immunoblots of N. caninum extracts. In addition, specific labeling of a 45 kDa band in Toxoplasma gondii tachyzoite extracts was observed. By immunofluorescence, these antibodies stained predominantly the apical portion of both N. caninum and T. gondii tachyzoites, but the protein was absent from the parasite surface. Immunogold localization in LR-White embedded N. caninum tachyzoites demonstrated staining of predominantly the apically located micronemes, as well as of dense granules located at the posterior end of the tachyzoites. As evidenced by immunohistochemistry, this Neospora microneme antigen and its immunoreactive counterpart in Toxoplasma appeared to be expressed in both tachyzoite and bradyzoite stages. PMID- 10802318 TI - Plasmodium vivax synonymous substitution frequencies, evolution and population structure deduced from diversity in AMA 1 and MSP 1 genes. AB - Polymorphic regions of the genes encoding Plasmodium vivax apical membrane antigen 1 (PvAMA1) and P. vivax merozoite surface protein 1 (PvMSP1) were sequenced to examine population diversity both within and between geographical areas. Sequences were obtained for 219 isolates for PvAMA1 and for 175 isolates for PvMSP1 from Africa, China, India, Indonesia, Philippines, Papua New Guinea, Solomon Islands and Thailand. Over half of the isolates were obtained from different regions within the Philippines, and this was used to look at the diversity within a country. Sixty nine haplotypes and 22 polymorphic sites in a 414-bp region of PvAMA1 and 41 haplotypes and 34 polymorphic sites in a 249-bp fragment of PvMSP1 were detected. For both PvAMA1 and PvMSP1, four previously unreported polymorphic nucleotide positions were identified. Population analysis indicated that there were significant differences in allele frequencies between different regions but these differences were small compared to the diversity within populations (Fixation index, F(ST), of 0.126 and 0.078 for PvAMA1 and PvMSP1, respectively). PvAMA1 and PvMSP1 had similar nonsynonymous substitution frequencies but surprisingly, the synonymous substitution frequency for PvMSP1 was eight times the frequency for PvAMA1 suggesting that synonymous substitutions in at least PvAMA1 are not neutral. PMID- 10802319 TI - Expression of Plasmodium falciparum trimeric G proteins and their involvement in switching to sexual development. AB - Both cholera and pertussis toxins were used to label and study the expression of heterotrimeric G protein alpha subunits in Plasmodium falciparum extracts. Expression of these proteins is developmentally regulated throughout the erythrocytic cycle with peak expression during early asexual development and in mature sexual stages. Treatment of P. falciparum cultures with cholera toxin causes an increase in conversion to sexual development, and at the same concentration has a marginal inhibitory effect on asexual growth and division. Through precise synchronisation of the parasites' asexual cell cycle, we have defined the period of sensitivity to this induction at around the time of invasion, one cycle before the development of the sexual form. Fluorescent microscopy confirmed that access of the toxin to the parasite is limited to the invasive form - the free merozoite, while further labelling studies revealed expression of a single G protein alpha subunit in these stages. These observations are consistent with the view that a G protein-dependent signal transduction pathway is involved in coupling the parasite's environment to commitment to sexual development (gametocytogenesis). This means of artificially stimulating the pathways leading to sexual development can now be used to biochemically follow the activation of the signalling pathways involved. PMID- 10802321 TI - Stage-specific development of a novel adenosine transporter in Leishmania donovani amastigotes. AB - Leishmania donovani, like all other kinetoplastida, is a purine auxotroph. Comparative studies of adenosine transport in L. donovani amastigotes and promastigotes revealed that, unlike the promastigote stage, the amastigote possesses two distinct adenosine transporters (T(1) and T(2)) both with high affinities (K(m), 1.14+/-0.05 and 2. 09+/-0.13 microM, respectively). One of these transporters (T(1)) appears to be identical with the adenosine/pyrimidine nucleoside transporter of the promastigote reported earlier. The other transporter (T(2)) is specific for the amastigote stage and transports only purine nucleosides. The biological significance of this stage-specific development of the second adenosine transporter has been briefly discussed. PMID- 10802320 TI - Identification of a novel antigenic domain of Plasmodium falciparum merozoite surface protein-1 that specifically binds to human erythrocytes and inhibits parasite invasion, in vitro. AB - Merozoite surface protein 1 (MSP-1) of Plasmodium falciparum is a promising candidate for vaccine development against malaria. Identification of protective epitopes within MSP-1 is an important step towards the elucidation of mechanisms of parasitic invasion and for the creation of a multi-subunit vaccine. In this study, we show that a 115 amino acid region (p115MSP-1) within the p38 domain of MSP-1 can: (i) specifically bind to human erythrocytes, independent of glycophorin A; (ii) inhibit parasite invasion at significant levels, in vitro; and (iii) be recognized by human sera of individuals from malaria-endemic regions of Africa. More importantly, we also show that polyclonal antibodies specific to this region prevent parasite invasion at levels approaching 90%, in vitro. Our data illustrate that not only is p115MSP-1 involved in parasite recognition/invasion of human erythrocytes, but that this region is highly antigenic, producing high titer antibodies. The delineation of the role of MSP-1 in parasite invasion is an important component of the development of a multi subunit malaria vaccine, and this study identifies a candidate antigen in this context. PMID- 10802322 TI - Cloning and expression of a gene encoding Sm16, an anti-inflammatory protein from Schistosoma mansoni. AB - The gene encoding Sm16, an anti-inflammatory, immunomodulatory protein present abundantly in secretions of the infective stages of Schistosoma mansoni was cloned and partially characterized. A data base analysis showed sequence homology to an earlier reported schistosomular stathmin-like gene sequences reported in dbEST and Genbank. The putative gene coding for Sm16 is of 500 bp with an open reading frame of 117 aa that included an N-terminal signal peptide sequence of 18 aa. There are three potential sites for phosphorylation (two serine and one tyrosine residue) but no glycosylation sites in the sequence. The coding region of Sm16 was amplified from cercarial cDNA, cloned and expressed in bacterial and insect expression systems. The purified recombinant protein showed strong immunoreactivity with a polyclonal rabbit anti-Sm16 antibody raised against the native anti-inflammatory protein Sm16. Contrary to earlier report, this gene appears to be not stage-specific. Metabolic labeling studies suggested that Sm16 is phosphorylated and is synthesized by both cercariae and schistosomula of S. mansoni. Sequence homology with human stathmin, a cell cycle regulatory phospho protein, was 30%. However, when probed with specific antibodies, no cross reactivity was observed between Sm16 and human stathmin. PMID- 10802323 TI - Conservation of calnexin in the early branching protozoan Tritrichomonas suis. AB - The cloning and characterization of Ts-p66, a calcium-binding protein representing calnexin of the protozoan parasite Tritrichomonas suis is described. A T. suis cDNA expression library was screened with monospecific antibodies affinity-purified on an immuno-reactive 66 kDa antigen in a Triton X-114 membrane protein fraction. The deduced amino acid sequence of the resulting cDNA clones revealed that Ts-p66 belongs to the calreticulin protein family and represents calnexin of T. suis. The key structural features and sequence motifs of the calnexins were all conserved. By lectin-blotting we demonstrated that the native protein is glycosylated. Northern and Southern hybridizations showed that T. suis calnexin was highly expressed and encoded by a single or low copy number gene. A cDNA encoding Ts-p66 was expressed as recombinant protein in Escherichia coli. By overlay with 45Ca it was demonstrated that the native and recombinant proteins bind Ca(2+). Using immunofluorescence with affinity-purified antibodies, a staining pattern was observed which points towards a putative localization of Ts p66 in the nuclear membrane and endoplasmic reticulum. Demonstration of a structurally conserved calnexin in the amitochondriate protist T. suis indicates the very early evolutionary origin of the machinery for quality control of protein folding in the endoplasmic reticulum and the molecules involved hereby. PMID- 10802324 TI - An Entamoeba histolytica inositol 1,3,4-trisphosphate 5/6-kinase has a novel 3 kinase activity. PMID- 10802325 TI - Vacuolar localization of an Entamoeba histolytica homologue of the plasma membrane ATPase (PMCA). PMID- 10802326 TI - Amplification of the ABC transporter gene PGPA and increased trypanothione levels in potassium antimonyl tartrate (SbIII) resistant Leishmania tarentolae. PMID- 10802327 TI - Ferredoxin involvement in metronidazole resistance of Giardia duodenalis. PMID- 10802328 TI - The role of potassium in the response of Giardia intestinalis to hypo-osmotic stress. PMID- 10802329 TI - Cell killing by the human parasite Entamoeba histolytica is inhibited by the rho inactivating C3 exoenzyme. PMID- 10802330 TI - Preface PMID- 10802331 TI - Epidemiologic issues in the validation of veterinary diagnostic tests. AB - In this review, we critically discuss the objectives, methods and limitations of different approaches for the validation of diagnostic tests. We show (based on published data and our own experiences) that estimates for the diagnostic sensitivity and specificity may vary among populations and/or subpopulations of animals, conditional on the distribution of influential covariates. Additional variability in those parameter estimates may be attributable to the sampling strategy. The uncertainty about diagnostic parameters is of concern for the decision-maker in the context of clinical diagnosis or quantitative risk assessment as well as for the epidemiologist who uses test data for prevalence estimation or risk-factor studies. Examples for the calculation of diagnostic parameters are presented together with bias-avoidance strategies. We suggest guidelines for an epidemiologic approach to test validation of veterinary diagnostic tests. PMID- 10802332 TI - Principles and practical application of the receiver-operating characteristic analysis for diagnostic tests. AB - We review the principles and practical application of receiver-operating characteristic (ROC) analysis for diagnostic tests. ROC analysis can be used for diagnostic tests with outcomes measured on ordinal, interval or ratio scales. The dependence of the diagnostic sensitivity and specificity on the selected cut-off value must be considered for a full test evaluation and for test comparison. All possible combinations of sensitivity and specificity that can be achieved by changing the test's cut-off value can be summarised using a single parameter; the area under the ROC curve. The ROC technique can also be used to optimise cut-off values with regard to a given prevalence in the target population and cost ratio of false-positive and false-negative results. However, plots of optimisation parameters against the selected cut-off value provide a more-direct method for cut-off selection. Candidates for such optimisation parameters are linear combinations of sensitivity and specificity (with weights selected to reflect the decision-making situation), odds ratio, chance-corrected measures of association (e. g. kappa) and likelihood ratios. We discuss some recent developments in ROC analysis, including meta-analysis of diagnostic tests, correlated ROC curves (paired-sample design) and chance- and prevalence-corrected ROC curves. PMID- 10802333 TI - Application of diagnostic tests in veterinary epidemiologic studies. AB - Important applications of diagnostic tests in non-clinical areas of veterinary medicine include surveillance, monitoring or screening for disease, prevalence estimation, and risk-factor studies. In this review, we address the processing and analysis of diagnostic data with special emphasis on serological data from enzyme-linked immunosorbent assays (ELISAs). We describe methods that are used to adjust for misclassification (imperfect sensitivity and specificity) in the different applications of diagnostic tests. These methods often have limitations attributable to the uncertainty of sensitivity and specificity estimates. We review descriptive and analytical statistical methods that allow adequate presentation of non-clinical diagnostic data. PMID- 10802334 TI - Estimation of sensitivity and specificity of diagnostic tests and disease prevalence when the true disease state is unknown. AB - The performance of a new diagnostic test is frequently evaluated by comparison to a perfect reference test (i.e. a gold standard). In many instances, however, a reference test is less than perfect. In this paper, we review methods for estimation of the accuracy of a diagnostic test when an imperfect reference test with known classification errors is available. Furthermore, we focus our presentation on available methods of estimation of test characteristics when the sensitivity and specificity of both tests are unknown. We present some of the available statistical methods for estimation of the accuracy of diagnostic tests when a reference test does not exist (including maximum likelihood estimation and Bayesian inference). We illustrate the application of the described methods using data from an evaluation of a nested polymerase chain reaction and microscopic examination of kidney imprints for detection of Nucleospora salmonis in rainbow trout. PMID- 10802335 TI - Herd-level interpretation of test results for epidemiologic studies of animal diseases. AB - Correct classification of the true status of herds is an important component of epidemiologic studies and animal disease-control programs. We review theoretical aspects of herd-level testing through consideration of test performance (herd level sensitivity, specificity and predictive values), the factors affecting these estimates, and available software for calculations. We present new aspects and considerations concerning the effect of precision and bias in estimation of individual-test performance on herd-test performance and suggest methods (pooled testing, targeted sampling of subpopulations with higher prevalence, and use of combinations of tests) to improve herd-level sensitivity when the expected within herd prevalence is low. PMID- 10802336 TI - Conditional dependence between tests affects the diagnosis and surveillance of animal diseases. AB - Dependence between the sensitivities or specificities of pairs of tests affects the sensitivity and specificity of tests when used in combination. Compared with values expected if tests are conditionally independent, a positive dependence in test sensitivity reduces the sensitivity of parallel test interpretation and a positive dependence in test specificity reduces the specificity of serial interpretation. We calculate conditional covariances as a measure of dependence between binary tests and show their relationship to kappa (a chance-corrected measure of test agreement). We use published data for toxoplasmosis and brucellosis in swine, and Johne's disease in cattle to illustrate calculation methods and to indicate the likely magnitude of the dependence between serologic tests used for diagnosis and surveillance of animal diseases. PMID- 10802337 TI - Log-linear and logistic modeling of dependence among diagnostic tests. AB - We developed log-linear and logistic-modeling approaches to investigate dependence among diagnostic tests. To illustrate the approaches, we used published data for swine toxoplasmosis, bovine paratuberculosis, and swine brucellosis. These diseases were selected because each animal's true disease status was known, at least five tests were used, and the serologic tests had been previously shown to have moderate-to-high pairwise dependence in test sensitivities (and sometimes in test specificities). Log-linear and logistic modeling yielded similar results for swine toxoplasmosis and swine brucellosis. However, logistic modeling could not be used to investigate test dependence for bovine paratuberculosis because of quasi-separation in the data attributable to two fecal-based tests having specificities of 100%. Findings from our modeling indicated that 3 (modified agglutination, enzyme-linked immunosorbent assay (ELISA), latex agglutination) of 5 serologic tests for toxoplasmosis and 2 (rivanol and particle concentration fluorescence immunoassay) of 6 serologic tests for brucellosis were adequate for diagnosis. For bovine paratuberculosis, both fecal-based tests (Herrold's egg-yolk culture and radiometric culture) and 1 (ELISA) of 3 serologic tests were necessary in serial and parallel testing schemes. PMID- 10802338 TI - Decision analysis: dealing with uncertainty in diagnostic testing. AB - Decision analysis is a process for systematically analyzing complex choices by considering all pertinent information. In this paper, we discuss how uncertainty associated with diagnostic testing can be included in a decision analysis using pay-off tables and decision trees (decision-flow diagrams). Variables associated with diagnostic test interpretation (such as pre-test and post-test probability of disease; test sensitivity, specificity and predictive values; fixed cut-offs versus continuous measurement scales; test dependence associated with the use of multiple tests) are considered. Several decision criteria and output measures are discussed (including MAXIMIN and MAXIMAX criteria, opportunity costs, expected monetary values, expected utility, sensitivity and risk-profile analysis, and threshold analysis). The application of decision analysis to diagnostic testing for Johne's disease and traumatic reticuloperitonitis of cattle, and for canine heartworm disease are used to illustrate both population- and patient-oriented applications and criteria for ranking the desirability of different outcomes. PMID- 10802339 TI - Cell cycle regulators in neural stem cells and postmitotic neurons. AB - In the mammalian central nervous system, neurons withdraw from the cell cycle immediately after their differentiation from proliferative neuroepithelial cells. Even while postmitotic neurons remain in permanent mitotic quiescence, they express a number of cell cycle regulators required for cell cycle progression. This review focuses on the expression and functions of members of the retinoblastoma protein (Rb) family (Rb, p107, p130) and necdin, all of which are growth suppressors that interact with the viral oncoproteins and the E2F family proteins. These molecules are differentially expressed in proliferative neural progenitors and postmitotic neurons in the developing neuroepithelium in vivo and differentiating embryonal carcinoma cells in vitro. During neurogenesis, dysfunction of the Rb family proteins causes impaired neuronal differentiation accompanied by cell death (apoptosis). Thus, the Rb family proteins are essential for both terminal mitosis of neuronal progenitors and survival of nascent neurons. However, the Rb family proteins seem to be dispensable for the maintenance of the postmitotic state of terminally differentiated neurons. Necdin is expressed exclusively in postmitotic cells and may contribute to their permanent mitotic arrest. These cell cycle regulators coordinately act in the generation, survival and demise of postmitotic neurons. PMID- 10802340 TI - Functions of glutamate transporters in the brain. AB - L-glutamate is the primary excitatory neurotransmitter in the mammalian central nervous system and has also been implicated as a potent neurotoxin. To ensure a high signal-to-noise ratio during synaptic transmission and to prevent neuronal damage that might occur as a result of excessive activation of glutamate receptors, the extracellular glutamate concentration is tightly controlled by glutamate transporters in the plasma membrane of neurons and the surrounding glial cells. Five subtypes of glutamate transporters have been identified and characterized by molecular cloning. Recent studies of glutamate transporters using the genetic knockout strategy indicate that glial, but not neuronal, glutamate transporters play critical roles in maintaining the extracellular glutamate concentrations and are thereby essential for both normal synaptic transmission at the photoreceptor synapses and protection of neurons against glutamate excitotoxicity. This review summarizes the current knowledge on the properties and functional roles of glutamate transporters, focusing on the properties of the anion channel in the transporters, the unexpected localization of these transporters, their role in synaptic transmission and plasticity, and their involvement in the protection of neurons against excitotoxicity. PMID- 10802341 TI - Fyn tyrosine kinase participates in the compact myelin sheath formation in the central nervous system. AB - The cellular mechanisms for spiral wrapping and compaction of myelin sheaths by oligodendrocytes are not known yet. In this study, we examined the role of fyn tyrosine kinase, which could be responsible for molecular events during the stage of myelination in the CNS. Western blot and immunohistochemical analyses revealed that fyn-deficient mice have significantly lower levels of myelin basic protein (MBP), which is required for intracellular membrane adhesion parts so-called major dense line (MDL) and thought to be essential for the stability of myelin sheath. Electron microscopy verified that the myelin ultrastructure could be used to distinguish fyn-deficient mice from wild-type mice, showing a thin and redundant myelin sheath in the corpus callosum. Further, the electron-dense 'major' line in myelin from the purified myelin fractions remained condensed, and myelin compaction was split opened in fyn-deficient mice. To determine whether there was a change in the microheterogeneity of MBP due to a post-translational event we first investigated peptidylarginine deiminase (PAD), which is an enzyme that converts arginine residues in peptides to citrulline residues. PAD immunoreactivity was observed both in the myelin from fyn-deficient and wild-type mice. By Western blot analysis we found an increase of the citrullined form of MBP. In addition, MBP from fyn-deficient mice did weakly induce vesicle aggregation properties of MBP-mediated adhesion. We concluded that although oligodendrocytes from fyn-deficient mice are able to wrap around the axon, they are unable to form compact myelin due to decreased MBP level and the presence of increased citrullinated MBP. PMID- 10802342 TI - Facilitation of neurotransmitter release at the spiny lobster neuromuscular junction. AB - Stimulation-induced facilitation of transmitter release was examined at spiny lobster (Palinurus japonicus) neuromuscular junctions by measuring excitatory junctional potentials (EJPs). We found three components of facilitation with the decay time constants about 16, 200 and 1000 ms, respectively. The decay time constants of the nerve terminal free Ca(2+) concentration after stimulation agreed well with the two slower time constants of facilitation. The relationship among the three components of facilitation and a yet slower component, S, was investigated on the basis of several models. The models that have a multiplicative relationship between any two components of facilitation, and the additive model between all components could not account for the results of experiments. Only the 'unified power model', which assumes that facilitation and S are described by the 3-4th power of the sum of underlying components, could account for both the growth process of EJPs during stimulation and the effects of Ca(2+)-chelators. Loading Ca(2+)-chelators, BAPTA and EGTA, into the presynaptic terminals resulted in reduction, but not elimination, of any component of the facilitation. These results suggest that in the spiny lobster neuromuscular junction, the 'unified power model' can describe the relationship between three components of facilitation and S, and that residual free Ca(2+) enhances all three components of facilitation, although it may not be essential for them. PMID- 10802343 TI - Laminar characteristics of functional connectivity in rat barrel cortex revealed by stimulation with caged-glutamate. AB - In rodent somatosensory (barrel) cortex input is processed by whisker-related columns before the integrated output is fed into behaviorally-relevant circuits. The layer-specific activation patterns of the rat barrel cortex were examined with a set-up for scanning functional connectivity in brain slices. Flash-induced release of caged-glutamate at a large number of stimulation sites was used in combination with simultaneous field potential recordings from layers II to VI with five electrodes. The field potentials revealed striking differences between the cortical layers. Glutamate-release in layer IV and lower layer III was most effective in evoking excitation in all other cortical layers, whereas field potentials recorded from layer IV itself were caused by stimulation of a very restricted columnar zone only. Field potentials in layers II and III were strongly driven by stimulation in layer IV and less consistently and much weaker by layer V. Layer V was the only lamina capable of responding to stimulation of all other cortical layers, thus displaying the largest input maps. Layer VI possessed functional connectivity intrinsically and with layer V. These data lead us to suggest that thalamic input may be boosted by its main target layer IV to start a sequence of excitation in layer IV, passing to the supragranular layers and finally reaching the infragranular layers. This sequence is likely to be backed-up by other simultaneous steps of transmission including a layer IV-to-V interaction. We proposed that the increasing size of the receptive fields when sampling granular, supragranular and infragranular layers in vivo, might have its functional basis in the laminar interactions described here in an in vitro preparation. PMID- 10802344 TI - BDNF rapidly induces aspartate release from cultured CNS neurons. AB - We investigated the effects of brain-derived neurotrophic factor (BDNF) on aspartate release from cultured cerebellar neurons. This release occurred within 1 min after the addition of 100 ng/ml BDNF. The amount of aspartate released was less than that of glutamate. Aspartate release induced by BDNF was rapid and transient, as in the case of glutamate. Although high potassium evoked the release of both excitatory (glutamate and aspartate) and inhibitory (GABA and glycine) amino acid transmitters, BDNF only induced glutamate and aspartate release. BDNF-induced aspartate release was completely blocked by pretreatment with K252a or TrkB-IgG. The aspartate release induced by BDNF was not dependent on the extracellular Ca(2+), but required intracellular Ca(2+) mobilization. These results showed that BDNF may be involved in excitatory transmission using aspartate as well as glutamate through TrkB-mediated signaling in cerebellum. PMID- 10802345 TI - Characterization of the subventricular zone of the adult human brain: evidence for the involvement of Bcl-2. AB - The subventricular zone (SVZ) is an embryonic remnant that persists and remains mitotically active throughout adulthood. The rodent SVZ harbors neuronal precursors, principally in its anterior part, and generates neuroblasts that migrate tangentially into the olfactory bulb, thus forming the so-called rostral migratory stream. This study aimed at characterizing the SVZ in the human brain. Antibodies raised against the widely used SVZ molecular markers nestin, glial fibrillary acidic protein, beta-tubulin-III and polysialylated neural cell adhesion molecule, have allowed us to characterize in detail a zone similar to the rodent SVZ in humans. Virtually all portions of the lateral ventricle, as well as the ventral (hypothalamic) sector of the third ventricle, displayed immunoreactivity for most of the molecular markers. The midline region of the septum (septal recess) and the ventral portion of the SVZ displayed a particularly intense immunostaining for all SVZ markers. These two regions may represent zones of adult neurogenesis that are unique to primates. Furthermore, the anti-apoptotic protein Bcl-2 was found to be actively synthesized and co expressed with all the other markers throughout the entire SVZ. This study reveals that a well-developed SVZ exists in the adult human brain and suggests that Bcl-2 might play an important role in the functional organization of such a system. PMID- 10802346 TI - Neither a general flexor nor a withdrawal pattern of nociceptive reflexes evoked from the human foot. AB - In humans motor reactions to noxious radiant heat stimulation of the sole and the dorsum of the foot do not resemble a locally specific pattern of multiple modular withdrawal reflexes but rather a general flexion reflex pattern with a few exceptions which did neither fit a withdrawal nor a flexion reflex pattern. The partly observed excitatory feed back to foot extensors from nociceptive afferents of the foot sole is functionally discussed as a foot stabilizing mechanism under particular conditions. PMID- 10802348 TI - RTOG 90-05: the real conclusion. PMID- 10802349 TI - American brachytherapy society recommendations for clinical implementation of NIST-1999 standards for (103)palladium brachytherapy. The clinical research committee of the American Brachytherapy Society. AB - PURPOSE: Recent important developments in palladium-103 ((103)Pd) dosimetry mandate a reevaluation of (103)Pd brachytherapy prescribing practices. METHODS AND MATERIALS: The clinical research committee of the American Brachytherapy Society (ABS) convened a consensus session of brachytherapists and physicists to develop recommendations regarding future dose prescribing guidelines for National Institute of Standards and Technology (NIST-1999) calibrated (103)Pd sources. RESULTS: The ABS recommends that clinicians attempt to reproduce the implant doses delivered and reported in the literature through the past decade. CONCLUSIONS: The following should be immediately implemented for (103)Pd dosimetry: 1) All practicing physicians, physicists, dosimetrists, and suppliers implement NIST-1999 air-kerma strength standard for (103)Pd brachytherapy. 2) All treatment planning systems and dose calculation algorithms must be updated to reflect new dose rate constants. The AAPM-recommended validated value for Theraseed model 200 is 0.665 cGy h(-1) U(-1). The dose rate constant for the Mentor MED3633 seed is currently reported as 0.68 cGy h(-1) U(-1). This latter value and the values for seeds from other manufacturers are awaiting independent confirmation. 3) Physicians who previously prescribed 115 Gy for (103)Pd monotherapy prostate implants should now prescribe 125 Gy. When using (103)Pd as a boost following 45 Gy of external beam irradiation, 100 Gy should be prescribed instead of the previous 90 Gy. It is critical that all three changes be implemented concurrently, because they are interdependent. PMID- 10802350 TI - TGF-beta1 and radiation fibrosis: a master switch and a specific therapeutic target? AB - Radiation fibrosis is a frequent sequel of therapeutic or accidental radiation overexposure in normal human tissues. One of the main fundamental problems yet unsolved in fibrotic tissues is the origin of the chronic activation of myofibroblasts within these tissues. It has been postulated that this chronic activation results from a continuous production of activating factors. In this context, fibrosis could be defined as a wound where continuous signals for tissue repair are emitted. Cytokines and growth factors probably play a central role in this process. Among them, transforming growth factor-beta1 (TGF-beta1) is considered as a master switch for the fibrotic program. This review discusses recent evidence on the critical role played by TGF-beta in the initiation, development, and persistence of radiation fibrosis. It summarizes the results concerning this factor after irradiation of various tissues and cells, with an emphasis on superficial fibrosis, including skin and subcutaneous tissues. Finally, recent data concerning the treatment of established fibrotic disorders of various etiology are presented, as well as the possible mechanisms involved in fibrosis regression, which show that the TGF-beta pathway may constitute a specific target for antifibrotic agents. PMID- 10802351 TI - Single dose radiosurgical treatment of recurrent previously irradiated primary brain tumors and brain metastases: final report of RTOG protocol 90-05. AB - PURPOSE: To determine the maximum tolerated dose of single fraction radiosurgery in patients with recurrent previously irradiated primary brain tumors and brain metastases. METHODS AND MATERIALS: Adults with cerebral or cerebellar solitary non-brainstem tumors /= 70) (p < 0.005) were independently associated with increased rates of 4-year survival. CONCLUSION: A high rate of complete radiographic response was observed following moderate doses of cranial irradiation (alone or in combination with CHOD chemotherapy). Although complete responders had a statistically significant survival advantage at 4 years when compared with partial responders and nonresponders, the majority of patients who achieved complete response were dead of disease by 4 years following treatment. Based on this analysis of the RTOG database, there is no rationale for radiation dose escalation as a therapeutic strategy to combat PCNSL. Consequently, the radiotherapy component of the current RTOG Phase II trial (RTOG 93-10) now includes relatively low total doses of hyperfractionated irradiation for patients without residual disease (36 Gy/1.2 Gy, twice a day) as well as a more aggressive chemotherapy regimen. PMID- 10802353 TI - Local and distant failures after limited surgery with positive margins and radiotherapy for node-negative breast cancer. AB - PURPOSE: To determine the outcome of patients with positive margins after lumpectomy for breast cancer and to address the issue of the relationship between local recurrences and distant metastasis in the absence of chemotherapy. METHODS AND MATERIALS: Among 3697 patients with primary breast cancer, we retrospectively analyzed 152 patients who had undergone conservative surgery with axillary dissection, had infiltrating carcinomas with positive margins, were node negative, and received radiotherapy without chemotherapy. One-third received hormonal therapy. Endpoints were local failure and distant metastasis. Median follow-up was 72 months. RESULTS: Five- and 10-year recurrence-free survival were 0.80 and 0.71 respectively for local recurrences, and 0.85 and 0.73 respectively for metastasis. Infiltrating carcinoma on the margins was associated with early local relapse as opposed to intraductal carcinoma. Local and distant recurrences had similar patterns of yearly-event probabilities. Hazard of relapsing from metastasis was 2.5 times higher after a local recurrence. In the multivariate analysis, negative estrogen receptors (ER-)(p = 0.0012), histologic multifocality (p = 0.0028), and no hormonal therapy (p = 0.017) predicted local relapses, while ER- (p = 0.004) and pathologic grade (p = 0.009) predicted metastasis. Hormonal therapy did not prevent early local recurrences. CONCLUSION: In this population, reexcision is advisable for local purposes and because the data support the hypothesis that local and distant recurrences are tightly connected. PMID- 10802354 TI - Radiographic evaluation of surgical clips is better than ultrasound for defining the lumpectomy cavity in breast boost treatment planning: a prospective clinical study. AB - PURPOSE: Breast radiotherapy (BrRT) often includes a "boost" to the lumpectomy bed. Selection of an appropriate electron energy and field size is based upon the dimensions and location of the surgical cavity. This study was performed to confirm our impression that ultrasound (US) inadequately defines the volume at risk compared to radiographic evaluation of surgical clips placed within the operative bed at the time of lumpectomy. METHODS AND MATERIALS Twenty-nine women treated with BrRT at our institution between 1996-1998 were prospectively identified as having surgical clips within the lumpectomy cavity. These patients underwent both US evaluation and radiographic evaluation of the clips to independently define two sets of transverse (T), longitudinal (L), and depth (D) measurements for each cavity. Volumes (V) were calculated for each method, assuming the cavity to be a box (V = T x L x D). Twenty-one women began BrRT following a median postoperative interval of 6 weeks (Group A), and 8 after 24 weeks (Group B) due to delivery of systemic therapy. RESULTS: Dimensions derived by US were smaller than the clip method in 22/29 of T, 25/29 of L, and 23/29 of D, or 80% of all linear measurements. A paired t-test demonstrated the difference between the methods to be statistically significant: T: p = 0.0004; L: p = 0.0001; D: p = 0.0004; and V: p = 0.0001. This underestimation by US did not fit any predictable pattern. Although the mean difference between methods (clips - US) was only 1.3, 1.9, and 1.1 cm for T, L, and D, respectively, differences ranged up to 5. 7, 9.2, and 5.5 cm for T, L, and D. The bias toward underestimation of V by US was significantly greater for Group B than Group A (p = 0. 03). CONCLUSIONS: US significantly underestimates all three dimensions of the lumpectomy cavity and hence the volume at risk compared to radiographic evaluation of surgical clips. Breast-conserving surgery should include placement of clips at the margins of the lumpectomy cavity (superior, inferior, medial, lateral, and posterior) to aid in radiotherapy treatment planning. US should not be utilized to guide the design of the boost field as it will result in inappropriate selection of lower electron energies and smaller field sizes (geographical miss), particularly among patients who receive chemotherapy prior to breast radiotherapy. PMID- 10802355 TI - Prone breast radiotherapy in early-stage breast cancer: a preliminary analysis. AB - PURPOSE: Women with large breasts have marked dose inhomogeneity and often an inferior cosmetic outcome when treated with breast conservation compared to smaller-sized patients. We designed a prone breast board, which both minimizes breast separation and irradiated lung or heart volume. We report feasibility, cosmesis, and preliminary local control and survival for selected women with Stage 0-II breast cancer. MATERIALS AND METHODS: Fifty-six patients with clinical Stage 0-II breast cancer were treated with lumpectomy and breast irradiation utilizing a prototype prone breast board. A total of 59 breasts were treated. Indications for treatment in the prone position were large or pendulous breast size (n = 57), or a history of cardiopulmonary disease (n = 2). The median bra size was 41D (range, 34D-44EE). Cosmesis was evaluated on a 1-10 (worst-to-best) scale. RESULTS: Acute toxicity included skin erythema (80% of patients experienced Grade I or Grade II erythema), breast edema (72% of patients experienced mild edema), pruritus (20% of patients), and fatigue (20% of patients reported mild fatigue). One patient required a treatment break. The only late toxicity was related to long-term cosmesis. The mean overall cosmesis score for 53 patients was 9.37 (range, 8-10). Actuarial 3- and 5-year local control rates are 98%. Actuarial overall survival at 3 and 5 years are 98% and 94%. CONCLUSION: Our data indicate that treating selected women with prone breast radiotherapy is feasible and tolerated. The approach results in excellent cosmesis, and short term outcome is comparable to traditional treatment techniques. This technique offers an innovative alternative to women who might not otherwise be considered candidates for breast conservation. PMID- 10802356 TI - Evaluation of a standard breast tangent technique: a dose-volume analysis of tangential irradiation using three-dimensional tools. AB - PURPOSE: A thorough dose-volume analysis of a standard tangential radiation technique has not been published. We evaluated the adequacy of a tangential radiation technique in delivering dose to the breast and regional lymphatics, as well as dose delivered to underlying critical structures. METHODS AND MATERIALS: Treatment plans of 25 consecutive women with breast cancer undergoing lumpectomy and adjuvant breast radiotherapy were studied. Patients underwent two-dimensional (2D) treatment planning followed by treatment with standard breast tangents. These 2D plans were reconstructed without modification on our three-dimensional treatment planning system and analyzed with regard to dose-volume parameters. RESULTS: Adequate coverage of the breast (defined as 95% of the target receiving at least 95% of the prescribed dose) was achieved in 16 of 25 patients, with all patients having at least 85% of the breast volume treated to 95% of the prescribed dose. Only 1 patient (4%) had adequate coverage of the Level I axilla, and no patient had adequate coverage of the Level II axilla, Level III axilla, or the internal mammary lymph nodes. CONCLUSION: Three-dimensional treatment planning is superior in quantification of the dose received by the breast, regional lymphatics, and critical structures. The standard breast tangent technique delivers an adequate dose to the breast but does not therapeutically treat the regional lymph nodes in the majority of patients. If coverage of the axilla or internal mammary lymph nodes is desired, alternate beam arrangements or treatment fields will be necessary. PMID- 10802357 TI - 3D conformal radiation therapy (3DCRT) for high grade prostate cancer: a multi institutional review. AB - PURPOSE: To evaluate the results of 3DCRT and the effect of higher than traditional doses in patients with high grade prostate cancer, we compiled data from three institutions and analyzed the outcome of this relatively uncommon subset of prostate cancer patients. METHODS AND MATERIALS: The 180 patients with Gleason score 8- 10 adenocarcinoma of the prostrate were treated with 3DCRT at the Univer sity of Michigan Health System, University of California-San Francisco, or Fox Chase Cancer. Eligible patients had T1-T4 NO or NX MO adenocarci noma with a pretreatment PSA. Pretreatment characteristics included: me dian age 72 years, 60.6% Gleason score 8 tumors, 57.6% T1-T2, and median pretreatment PSA 17.1 ng/ml (range 0.3-257.1). The total dose received was <70 Gy in 30%, 70-75 Gy in 37%, and >75 Gy in 33%, 27% received adju vant or neoadjuvant hormonal therapy. The median follow-up was 3.0 years for all patients and 16% of patients were followed up for at least 5 years. RESULTS: The 5-year freedom from PSA failure was 62.5% for all patients and 79.3% in T1-T2 patients. Univariate analysis revealed that T-stage (T1-T2 vs. T3-T4), pretreatment PSA, and RT dose predicted for freedom from PSA failure. A 5-year overall survival for all patients was 67.3%. Only RT dose was predictive of 5-year overall survival on univariate analysis. Because a significant association was seen between T-stage and RT dose, the Cox proportional hazards model was performed separately for T1 T2 and T3-T4 tumors. None of the prognostic factors reached statistical significance for overall survival or freedom from PSA failure in T3-T4 patients or for overall survival in T1-T2 patients. Lower RT dose and higher pretreatment PSA predicted for PSA failure on multivariate analysis in T1-T2 patients. CONCLUSION: This retrospective study from three institutions with experience in dose escalation suggests a dose effect for PSA control above 70 Gy in patients with T1-T2 high grade prostate cancer. These results are superior to surgery and emphasize the need for dose escalation in treating Gleason 8-10 prostate cancer. PMID- 10802358 TI - Interim report of image-guided conformal high-dose-rate brachytherapy for patients with unfavorable prostate cancer: the William Beaumont phase II dose escalating trial. AB - PURPOSE: We analyzed our institution's experience treating patients with unfavorable prostate cancer in a prospective Phase II dose-escalating trial of external beam radiation therapy (EBRT) integrated with conformal high-dose-rate (HDR) brachytherapy boosts. This interim report discusses treatment outcome and prognostic factors using this treatment approach. METHODS AND MATERIALS: From November 1991 through February 1998, 142 patients with unfavorable prostate cancer were prospectively treated in a dose-escalating trial with pelvic EBRT in combination with outpatient HDR brachytherapy at William Beaumont Hospital. Patients with any of the following characteristics were eligible: pretreatment prostate-specific antigen (PSA) >/= 10.0 ng/ml, Gleason score >/= 7, or clinical stage T2b or higher. All patients received pelvic EBRT to a median total dose of 46.0 Gy. Pelvic EBRT was integrated with ultrasound-guided transperineal conformal interstitial iridium-192 HDR implants. From 1991 to 1995, 58 patients underwent three conformal interstitial HDR implants during the first, second, and third weeks of pelvic EBRT. After October 1995, 84 patients received two interstitial implants during the first and third weeks of pelvic EBRT. The dose delivered via interstitial brachytherapy was escalated from 5.50 Gy to 6.50 Gy for each implant in those patients receiving three implants, and subsequently, from 8.25 Gy to 9.50 Gy per fraction in those patients receiving two implants. To improve implant quality and reduce operator dependency, an on-line, image-guided interactive dose optimization program was utilized during each HDR implant. No patient received hormonal therapy unless treatment failure was documented. The median follow-up was 2.1 years (range: 0.2-7.2 years). Biochemical failure was defined according to the American Society for Therapeutic Radiology and Oncology Consensus Panel definition. RESULTS: The pretreatment PSA level was >/= 10.0 ng/ml in 51% of patients. The biopsy Gleason score was >/= 7 in 58% of cases, and 75% of cases were clinical stage T2b or higher. Despite the high frequency of these poor prognostic factors, the actuarial biochemical control rate was 89% at 2 years and 63% at 5 years. On multivariate analysis, a higher pretreatment PSA level, higher Gleason score, higher PSA nadir level, and shorter time to nadir were associated with biochemical failure. In the entire population, 14 patients (10%) experienced clinical failure at a median interval of 1.7 years (range: 0.2 4.5 years) after completing RT. The 5-year actuarial clinical failure rate was 22%. The 5-year actuarial rates of local failure and distant metastasis were 16% and 14%, respectively. For all patients, the 5-year disease-free survival, overall survival, and cause-specific survival rates were 89%, 95%, and 96%, respectively. The 5-year actuarial rate of RTOG Grade 3 late complications was 9% with no patient experiencing Grade 4 or 5 acute or late toxicity. CONCLUSION: Pelvic EBRT in combination with image-guided conformal HDR brachytherapy boosts appears to be an effective treatment for patients with unfavorable prostate cancer with minimal associated morbidity. Our dose-escalating trial will continue. PMID- 10802359 TI - Urinary morbidity with a modified peripheral loading technique of transperineal (125)i prostate implantation. AB - PURPOSE: Analysis of urinary morbidity within the first 12 months following a modified peripheral loading technique for permanent transperineal transrectal ultrasound (TRUS) guided (125)I prostate implantation and comparison of urinary morbidity with various clinical and implant parameters. MATERIALS AND METHODS: Between October 1, 1996, and March 11, 1998, 87 patients with favorable, early stage prostate cancer were treated with permanent transperineal TRUS guided (125)I prostate implantation. A peripheral loading technique was utilized for source placement with 75-80% source distribution in the periphery and 20-25% source distribution centrally. A mean total activity of 38 mCi of (125)I was implanted (range, 19-66 mCi). The mean source activity was 0.43 mCi/source (range, 0.26-0.61 mCi/source) and the mean number of sources implanted was 88 (range, 56-134). The minimum prescribed dose to the prostate was 145 Gy. The median D(90), V(100), and V(150) were 152 Gy (range, 104-211 Gy), 92% (range, 71 99%), and 61% (range, 11-89%), respectively. The median follow-up time was 19 months (range, 12-29 months). Urinary morbidity was scored at 3 weeks and then at 3-month intervals for the first 2 years using a modified Radiation Therapy Oncology Group (RTOG) grading system (scale 0-5). RESULTS: Most patients developed at least minor urinary symptoms with frequency or nocturia being the most common. Overall, 79% (69/87) of patients experienced urinary morbidity with 21% (18/87) reporting no symptoms. The incidence of overall Grade 1 urinary morbidity was 37% (32/87); Grade 2 morbidity was 37% (32/87); and Grade 3 morbidity was 6% (5/87). There was no Grade 4 or 5 morbidity. The incidence of Grade 0 frequency/nocturia was 36% (31/87); Grade 1 was 33% (29/87); Grade 2 was 30% (26/87); and Grade 3 was 1% (1/87). Grade 0 dysuria was seen in 56% (49/87) of patients; 32% (28/87) had Grade 1; 10% (9/87) Grade 2; and 1% (1/87) Grade 3 dysuria. Most urinary symptoms started a few weeks after implantation and began to subside by 6 months. At 12 months, 22% (19/87) of patients had persistent urinary symptoms (78% Grade 0, 15% Grade 1, 3% Grade 2, and 3% Grade 3). The mean urethral point dose was 174 Gy (range, 99-315 Gy). The mean number of sources implanted correlated significantly with the likelihood of developing acute urinary morbidity (p = 0.03). The total activity implanted also correlated with the morbidity outcome dysuria (p = 0.01) with a threshold seen at 37 mCi. Urethral point dose, source activity, intraoperative TRUS prostate volume, D(90), V(100), V(150), patient age, pretreatment PSA, Gleason score, and T stage did not correlate with morbidity. CONCLUSIONS: Permanent transperineal TRUS guided (125)I prostate implantation using a modified peripheral loading technique is associated with mild urinary morbidity that resolves in 78% of patients by 12 months. Grade 3 urinary morbidity was encountered in only 6% (5/87) of patients. Urinary morbidity may be related to the total number of sources implanted and/or the total activity implanted. Overall urinary morbidity was not correlated with urethral point dose, source activity, intraoperative TRUS prostate volume, D(90), V(100), V(150), patient age, pretreatment PSA, Gleason score, and T stage. The low incidence of urinary morbidity may be a consequence of our modified peripheral loading technique and/or the selection of patients with good-to excellent preimplant urological parameters. Longer follow-up is necessary to assess biochemical control rates and long-term morbidity. PMID- 10802360 TI - Effects of pelvic rotation and needle angle on pubic arch interference during transperineal prostate implants. AB - PURPOSE: Pubic arch interference due to an enlarged prostate gland or a narrow pubic arch is often a limiting factor in adequate prostate coverage during transperineal brachytherapy. The purpose of this study was to evaluate the effects of both pelvic rotation and needle angles on pubic arch interference using CT-based 3-D information. METHODS AND MATERIALS: Seven patients had CT imaging in both supine and lithotomy positions and 3-D treatment planning was performed with three needle angles (20 downward, 0, 20 upward). The pubic arch interference was then measured and comparisons were made for each needle trajectory and pelvic position. RESULTS: Increasing pelvic rotation from supine to lithotomy position shows less pubic arch interference. Directing the needle tip upward shows less pubic arch interference in both supine and lithotomy positions when compared to needle tips directed downward. CONCLUSIONS: Both pelvic position and needle angles are important factors influencing pubic arch interference. Preplanning CT-based 3-D information may assist for individualized treatment planning in patients with a significant bony interference, thus avoiding pubic arch interference during implantation. PMID- 10802361 TI - Patient positioning in prostate radiotherapy: is prone better than supine? AB - PURPOSE: To assess potential dose reductions to the rectum and to the bladder with three-dimensional conformal radiotherapy (3D-CRT) to the prostate in the prone as compared with the supine position; and to retrospectively evaluate treatment position reproducibility without immobilization devices. METHODS AND MATERIALS: Eighteen patients with localized prostate cancer underwent pelvic CT scans and 3D treatment planning in prone and supine positions. Dose-volume histograms (DVHs) were constructed for the clinical target volume, the rectum and the bladder for every patient in both treatment positions. "Comparative DVHs" (cDVHs) were defined for the rectum and for the bladder: cDVH was obtained by subtracting the organ volume receiving a given dose increment in the prone position from the corresponding value in the supine position. These values were then integrated over the entire dose range. The prescribed dose to the planning target volume (PTV) was 74 Gy using a 6-field technique. To evaluate reproducibility, portal films were subsequently reviewed in 12 patients treated prone and 10 contemporary patients treated supine (controls). No immobilization devices were used. Deviations in the anterio-posterior (X) and cranio-caudal (Y) axes were measured. Mean treatment position variation, total setup variation, systematic setup variation, and random setup variation were obtained. RESULTS: Prone position was associated with a higher dose to the rectum or to the bladder in 6 (33%) and 7 (39%) patients, respectively. A simultaneously higher dose to rectum and bladder was noted in 2 (11%) patients in prone and in 7 (39%) patients in supine. Rectal and bladder volumes were frequently larger in prone than in supine: mean prone/supine volume ratios were 1.21 (SD, 0.68) and 1.03 (SD, 1.32), respectively. In these cases cDVH analysis more often favored the prone position. Mean treatment position variation and total setup variation were similar for both prone and supine plans. A higher systematic setup variation was observed in prone positioning: 2.7 mm vs. 1.9 mm (X axis) and 4.1 mm vs. 2.2 mm (Y axis). The random variation was similar for both prone and supine: 4. 0 mm vs. 3.6 mm (X axis) and 3.7 mm vs. 3.6 mm (Y axis). CONCLUSIONS: Prone position 3D-CRT is frequently, but not always, associated with an apparent dose reduction to the rectum and/or to the bladder for prostate cancer patients. As suggested by the increased mean prone/supine rectal volume ratio, the advantage of prone positioning for the rectum may be artifactual, at least partly reflecting a position-dependent rectal air volume, which may significantly vary from treatment to treatment. In the absence of immobilization devices, daily setup reproducibility appears less accurate for the prone position, primarily due to systematic setup variations. PMID- 10802362 TI - Chemoradiotherapy for muscle invading bladder carcinoma. Final report of a single institutional organ-sparing program. AB - PURPOSE: Chemoradiotherapy is becoming an alternative to radical cystectomy among patients with muscle invading bladder cancer. We began a prospective study in 1988 to determine the possibilities of conservative treatment and aiming to improve the results obtained by cystectomy alone in invasive bladder cancer. A combination of methotrexate, vinblastine, adriamycin, and cisplatin (M-VAC), followed by radiotherapy and concomitant cisplatin was used. METHODS: Fifty patients with good performance status and with stages T2 to T4 operable untreated invasive bladder cancer were entered in the study. Treatment protocol was as follows: (i) cytoreductive transurethral resection; (ii) two cycles of M-VAC chemotherapy; (iii) radiotherapy, 45 Gy on pelvic volume and, at the same time, 20 mg/m(2) cisplatin on days 1 to 5. Cystoscopic evaluation: if there was a complete response, radiotherapy was completed up to 65 Gy; if there was not a complete response, a cystectomy was performed. Median follow-up of the series was 73 months (18-180 m). RESULTS: Tumor response was as follows: 34 complete responses (68%), 9 partial responses (18%), and 7 nonresponses (14%) were observed. The 5-year overall survival and local control were 48% and 47%, respectively. For the complete responder patient, 5-year survival and local control were 65% and 70%, respectively. Severe toxicity was uncommon. The most frequent were leucopenia and cystitis. No treatment-related deaths occurred with either treatment protocol. CONCLUSIONS: Conservative combination treatment may be an acceptable alternative to immediate cystectomy in selected patients with bladder cancer, although a randomized clinical trial would be required to produce definitive results. PMID- 10802363 TI - A randomized trial of hypofractionated schedules of palliative radiotherapy in the management of bladder carcinoma: results of medical research council trial BA09. AB - PURPOSE: To compare the efficacy and toxicity of two hypofractionated radiotherapy schedules for the improvement of local symptoms from muscle-invasive bladder cancer. METHODS AND MATERIALS: A multicenter randomized trial was conducted comparing the efficacy and toxicity of two radiotherapy schedules (35 Gy in 10 fractions and 21 Gy in 3 fractions) for symptomatic improvement in patients considered unsuitable for curative treatment through disease stage or comorbidity. The primary outcome measures were overall symptomatic improvement of bladder-related symptoms at 3 months and changes in bladder- and bowel-related symptoms from pretreatment to end-of-treatment and 3-month assessments. Overall symptomatic improvement was defined prospectively as the improvement in one bladder-related symptom of at least one grade at 3 months, with no deterioration in any other bladder-related symptom. RESULTS: Five hundred patients were recruited, but data on symptomatic improvement at 3 months was only available on 272 patients. Of these, 68% achieved symptomatic improvement (71% for 35 Gy, 64% for 21 Gy), with no evidence of a difference in efficacy or toxicity between the two arms. There was no evidence of a difference in survival between the two schedules (hazard ratio [HR] = 0.99, 95% CI 0.82-1.21, p = 0. 933). CONCLUSION: This is the largest prospective trial to date in the palliative treatment of bladder cancer, and provides baseline data against which other results may be compared. The use of 21 Gy in 3 fractions appears as effective as 35 Gy in 10 fractions, although modest differences in survival, symptomatic improvement rates, and toxicity can not be reliably excluded. PMID- 10802364 TI - The influence of beam energy on the outcome of postoperative radiotherapy in head and neck cancer patients: secondary analysis of RTOG 85-03. AB - PURPOSE: To determine whether any difference in toxicity or efficacy occurs when head and neck cancer patients are treated postoperatively with (60)C0, 4 MV, or 6 MV photon beam. METHODS AND MATERIALS: This is a secondary analysis of the Intergroup Study 0034. Three hundred ninety-two patients were evaluable for comparison between treatment with (60)C0, 4 MV, or 6 MV photon beam. All patients had advanced but operable squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or larynx. Patients were randomized following surgical resection to receive treatment with either postoperative irradiation alone, or postoperative irradiation plus three cycles of cisplatin and 5-fluorouracil. Patients were categorized as having either "low risk" or "high risk" treatment volumes based on whether the surgical margin was 5 mm or less, presence of extra capsular nodal extension, and/or carcinoma in situ at the surgical margins. Low-risk volumes received 50-54 Gy, and high-risk volumes were given 60 Gy. Patients were compared in regards to acute and late radiotherapy toxicities as well as overall survival and loco-regional control according to the beam energy used. RESULTS: One-hundred fifty-seven, 140, and 95 patients were treated by (60)C0, 4 MV, and 6 MV, respectively. No differences were seen in acute or late toxicity among treatment groups. Locoregional control was achieved in 75%, 79%, and 80% of patients treated with (60)C0, 4 MV, or 6 MV (p = 0.61). Patients treated with 6 MV had a higher incidence of ipsilateral neck failure as first event (13%) than patients treated by (60)C0 and 4 MV (9%). This difference was not statistically significant. CONCLUSION: No differences in outcome, acute, or late toxicity were discernible in patients with advanced head and neck cancer treated with (60)C0, 4 MV, or 6 MV. This result should be interpreted with caution as increased incidence, albeit nonsignificant, of ipsilateral neck recurrence was observed in patients treated with 6 MV and the power of the study to detect a statistically significant difference is small. PMID- 10802365 TI - Skull-base invasion of nasopharyngeal carcinoma: magnetic resonance imaging findings and therapeutic implications. AB - PURPOSE: To evaluate the value of skull-base abnormality on MRI for predicting local recurrence in nasopharyngeal carcinoma. MATERIALS AND METHODS: Between November 1988 and February 1997, 48 patients with NPC were examined with both MRI (1.5 T) and CT prior to radiation therapy. T classification (1987 UICC) based on physical examination and CT findings were T1 in 3 cases, T2 in 22, T3 in 9, and T4 in 14. On MRI, low-intensity tissue with Gd enhancement in the marrow of the skull was considered to be a suspicious finding of skull-base invasion. CT simulation was performed in all patients. The total dose to the primary tumor was 60-75 Gy (mean, 67 Gy). The mean follow-up period was 42 months. RESULTS: All 14 T4 patients had abnormal tissue in the marrow of the skull base on MRI. Thirty eight percent (13 of 34) of T1-3 patients were suspected to have skull-base invasion based on MRI (0% for T1, 27% [6 of 22] for T2, and 78% [7 of 9] for T3). The 5-year local control rate was significantly different between T1-3 and T4 tumors (97% vs. 69%, p < 0.025) but was not different by the presence of the MRI abnormality in the skull base. CONCLUSION: Skull-base invasion suspected solely by MRI does not relate to local recurrence provided that careful treatment planning is performed with the aid of MRI and CT simulator. PMID- 10802366 TI - Locally advanced nasopharyngeal carcinoma: computed tomography findings, clinical evaluation, and treatment outcome. AB - PURPOSE: We present our experience with computed tomography (CT) for delineating the extent of bone erosion in nasopharyngeal carcinoma (NPC) and propose that a new subdivision of Stage T4 disease be added to the staging criteria for cases of minimal bone disease, defined as erosion of the base of the sphenoid or the pterygoid without cranial nerve (CN) involvement. METHODS AND MATERIALS: We retrospectively reviewed the clinical findings, radiological findings, and treatment outcome in 64 patients with Stage T4 NPC, diagnosed according to the American Joint Committee on Cancer 4th edition criteria. The median follow-up was 34 months (range, 3-118 months). Statistical analyses were performed using the chi-square test, the Kaplan-Meier method, and the log-rank test. RESULTS: Local control was achieved in 19 (46%) of 41 patients with CN deficits and 18 (78%) of 23 patients without CN deficits (p = 0.01). Overall 5-year survival with and without CN deficits was 25% and 58%, respectively (p = 0.01). When the 16 patients with minimal bone disease were compared to the remaining 48 patients, there were significant differences in local control rates (87% vs. 48%, p = 0. 006) and 5-year survival rates (68% vs. 28%, p = 0.008). CONCLUSION: Among patients with Stage T4 NPC, a subgroup of patients with only minimal bone disease may have a more favorable prognosis, which may have a considerable bearing on our approach to this patient group. PMID- 10802367 TI - Salvage brachytherapy for patients with locally persistent nasopharyngeal carcinoma. AB - PURPOSE: Locally persistent nasopharyngeal carcinoma (NPC) carries an increased risk of local failure if additional treatment is not given. This study was conducted to evaluate the outcomes of patients with locally persistent NPC as treated by high-dose-rate (HDR) intracavitary brachytherapy, and to explore whether routine brachytherapy boost could improve the local control. METHODS AND MATERIALS: Eighty-seven patients with locally persistent NPC treated during 1990 1998 with HDR intracavitary brachytherapy were retrospectively analyzed. Fibreoptic nasopharyngoscopy was performed 3-6 weeks after completion of the primary external radiation therapy (ERT). Biopsies were only taken from suspicious areas. Those with complete regression of local disease were put on observation. Eighty-seven patients were shown to have persistent viable disease at a median time of 6 weeks post-RT. The distribution according to Ho's staging system at initial diagnosis was as follows: Stage I-8, II-33, III-41, IV-5; T1 19, T2-48, T3-20; N0-32, N1-22, N2-28, N3-5. CT scan for restaging was not performed after the documentation of persistent disease. Our policy was to treat all patients with persistent disease with brachytherapy irrespective of the extent of disease just prior to brachytherapy. They were treated with HDR intracavitary brachytherapy, with either cobalt sources or an iridium source, giving 22.5-24 Gy in 3 weekly sessions in all but 4 patients. This dose was prescribed at a distance of 1.5 cm from the center of the surface as defined by the sources in the first six patients and subsequently reduced to 1 cm for the others. Twelve patients were treated with neoadjuvant chemotherapy. To compare the efficacy of brachytherapy, another 383 consecutive nonmetastatic patients, treated with curative intent by ERT, during the years 1990-1993, were evaluated. Multivariate analysis was performed using the Cox regression proportional hazards model. RESULTS: The 5-year actuarial local failure-free survival (LFFS) rates and disease-specific survival rates for the brachytherapy group and ERT group were 85% and 76.6% (p = 0.15), and 72% and 67.8% (p = 0.2), respectively. The corresponding 5-year actuarial LFFS rates for T1, T2, and T3 disease were 94.7%, 88.2%, 67.4%, and 84.1%, 79.8%, 62.6%. In assessing the local control, only the T staging was significant on multivariate analysis (p = 0.0004). Other parameters such as age, sex, and persistence of disease (giving brachytherapy) were all nonsignificant. Complications were comparable between the two groups. In the persistent group, the local failure rates of the patients treated with and without neoadjuvant chemotherapy were 17% (2/12) and 13% (10/75) respectively. When analyzed according to different brachytherapy sources, the 5-year LFFS rates of the T1, T2, and T3 patients treated with iridium and cobalt sources were 100% vs. 85.7 (p = 0.19), 93.6% vs. 70% (p = 0.04), and 67.7% vs. 60% (p = 0.72). The difference was statistically significant for the T2 groups. When early T-stage (T1 and T2) patients were grouped together for analysis, the iridium group again showed a statistically significant improvement in 5-year LFFS rate when it was compared with the cobalt group (95.3% vs. 76.5%, p = 0.03) and the ERT alone group (95.3% vs. 81.5%, p = 0.03). The improvement of local control is attributed to a higher nasopharyngeal mucosal dose that is achieved by using small-size flexible applicators with an iridium source. It is interesting to note that the 5 year LFFS rates for the ERT alone group (T1: 84.1%, T2: 79.8%, T3: 62.6%) are comparable to the corresponding rates of the cobalt group. This information supports our speculation that an adequate booster treatment could compensate for inadequate primary treatment. The prognosis of patients with locally recurrent NPC is grave. Maximizing the local control is therefore crucial for the survival of the patients. (ABSTRACT TRUNCATED) PMID- 10802368 TI - Concomitant boost radiation therapy for inoperable non-small-cell lung cancer: preliminary report of a prospective randomized study. AB - PURPOSE: The radiation therapy results for patients with inoperable non-small cell lung cancer (NSCLC) have been disappointing. Tumor dose escalation using concomitant boost technique (CBT) has been shown to improve local control in a few prospective studies. This trial was carried out to prospectively assess the radiation response and acute toxicity of CBT in comparison to the conventional treatment technique (CTT). METHODS AND MATERIALS: Ninety-seven consecutive eligible patients were entered in this prospective clinical trial between November 1994 and February 1998. Patients were randomized to receive either CBT (43 patients) or CTT (54 patients) radiation therapy. These patients either refused chemotherapy or were judged as unsuitable for chemotherapy. Patients in the CBT group received 46.8 Gy in 26 fractions using large fields that encompassed the gross and occult disease. A concomitant boost of 18.2 Gy (0.7 Gy per fraction) was delivered to the gross disease using small fields with 1.5-cm margins. The small fields were treated concurrently with the large fields and the total dose to the tumor area was 65 Gy in 26 fractions. Patients in the CTT group received 70.8 Gy in 38 fractions. The acute toxicity between each group was compared. The response rate was analyzed and compared by treatment group, gender, age, stage, histology, initial Karnofsky performance score (KPS), severity of acute toxicity, and maximum body weight loss (MBWL) during treatment course. RESULTS: The demographic parameters such as sex, age, and stage were evenly distributed in each treatment group. The majority of these patients had Stage IIIA and IIIB disease. Overall median treatment times were 39 days for the CBT group of patients and 62 days for the CTT group. No treatment-related mortality was found. There were 2 patients in the CTT group with acute RTOG Grade 3 lung toxicity, and no Grade 3 lung or esophageal toxicity was observed in CBT group. The response rates, assessed by radiographic images, were 69.8% and 48.1% for the CBT and CTT patients, respectively. Univariate and multivariate analysis revealed that patients in the CBT group, patients with better KPS, and patients with more severe acute toxicity had a higher response rate. CONCLUSION: This study demonstrates that concomitant boost radiation therapy is tolerable, and produces a superior response rate than conventional radiation therapy for patients with inoperable NSCLC. The length of treatment was reduced from 38 to 26 treatment days, almost a 30% reduction. PMID- 10802369 TI - Outcome of patients receiving photodynamic therapy for early esophageal cancer. AB - PURPOSE: Photodynamic therapy (PDT) has shown remarkable activity in a variety of human cancers. In the present study, we report the effects of PDT on inoperable early-stage esophageal cancer. METHODS AND MATERIALS: Sixty-two patients were treated with an argon dye laser (630 nm wavelength, 300-800 mW of power, energy dose of 200-300 J/cm) after intravenous injection of 5 mg/kg of hematoporphyrin derivative. Eighteen patients (29.5%) had in situ carcinoma (Tis), 30 (48.5%) had T1-stage cancer, 7 (11%) had T2-stage cancer, and 7 (11%) had recurrent disease in the anastomotic area after previous surgery without evidence of invasion outside the lumen. Patients with residual disease after two rounds of PDT received definitive radiotherapy. Patients were evaluated for response to therapy and survival. The follow-up time ranged from 3 to 90 months (median, 32 months). RESULTS: The complete response (CR) rate was 37% (23 of 62) in patients who received PDT alone and 82% (51 of 62) in those who also received radiotherapy. The CR rate after PDT alone was statistically higher (p = 0.04) for patients who had Tis/T1 lesions (21 of 48; 44%) than for those with T2-stage disease (2 of 7; 28%) or recurrent tumors (0 of 7; 0%). Fifty-two percent of patients who had CR following PDT alone did not suffer local tumor recurrence. The median local progression-free survival times after PDT and additional radiotherapy (in cases with incomplete response) was 49 months for Tis- and T1-stage lesions, 30 months for those with T2-stage disease, and 14 months for patients with locally recurrent disease. Patients who completely responded to PDT had a median overall survival (OS) of 50 months, which was significantly longer (p < 0.003) than that of patients not responding to PDT. Toxicity was minimal; we recorded three cases of esophageal stenosis (7%) and one case of tracheo-esophageal fistula (2.5%) after combined PDT and radiotherapy. CONCLUSION: PDT is an effective regimen for early esophageal cancer, giving a CR rate of about 40%, long-term local control and favorable overall survival. Additional radiotherapy in cases of incomplete response to PDT is effective and potentially curative in another 45% of cases. PMID- 10802370 TI - Feasibility and first results of multimodality treatment, combining EBRT, extensive surgery, and IOERT in locally advanced primary rectal cancer. AB - PURPOSE: To assess the outcome of aggressive multimodality treatment with preoperative external beam radiation therapy (EBRT), extended circumferential margin excision (ECME) and intraoperative electron beam radiation therapy (IOERT) in patients with locally advanced primary rectal cancer. METHODS AND MATERIALS: Thirty-eight patients with primary locally advanced rectal cancer, but without distant metastases, received multimodality treatment. CT-scan showed extension to other structures in 15 patients (39%) and definite infiltration into the surrounding structures in 23 patients (61%). All patients received preoperative EBRT (dose range 25-61 Gy) and 82% received 50.4 Gy. The resection types were: 12 low anterior resections (31%), 14 abdomino-perineal resections (37%), 6 abdomino transsacral resections (16%), and 6 pelvic exenterations (16%). The IOERT dose ranged from 10 to 17.5 Gy depending on the completeness of the resection. RESULTS: There was no perioperative mortality. The resection margins were microscopically negative in 31 patients (82%), microscopically positive in 4 (10%), and positive with gross residual disease in 3 patients (8%). Pelvic recurrences were observed in 5 patients (13%) including 3 IOERT infield failures. The overall 3-year local control, disease-free survival (DFS), and survival rates were 82%, 65%, and 72%, respectively. Negative resection margins were the most significant prognostic factor with regard to DFS (p = 0.0003) and distant control (p = 0.002) compared with cancer involved surgical margins. CONCLUSION: A high percentage of curative resections can be achieved in this group of patients with locally advanced rectal cancers. Adding IOERT to preoperative EBRT and ECME achieves high local control rates and possibly improves survival. PMID- 10802371 TI - Local radiotherapy with or without transcatheter arterial chemoembolization for patients with unresectable hepatocellular carcinoma. AB - PURPOSE: To evaluate the treatment outcome, patterns of failure, and prognostic factors for patients with unresectable hepatocellular carcinoma (HCC) treated with local radiotherapy alone or as an adjunct to transcatheter arterial chemoembolization (TACE). METHODS AND MATERIALS: From March 1994 to December 1997, 25 patients with unresectable HCC underwent local radiotherapy to a portion of the liver. Twenty-three patients were classified as having cirrhosis in Child Pugh class A and 2 in class B. Mean diameter of the treated hepatic tumor was 10.3 cm. Mean dose of radiation was 46.9 +/- 5.9 Gy in a daily fraction of 1.8-2 Gy. Sixteen patients were also treated with Lipiodol and chemotherapeutic agents mixed with Ivalon or Gelfoam particles for chemoembolization, either before and/or after radiotherapy. Percutaneous ethanol injection therapy (PEIT) was given to one patient. All patients were monitored for treatment-related toxicity and for survival and patterns of failure. RESULTS: In a median follow-up period of 23 months, 11 patients were alive and 14 dead. The median survival duration from treatment was 19.2 months with a 2-year survival of 41%. Only 3 of 25 patients had local progression of the treated hepatic tumor. The recurrences were seen within the liver or extrahepatic. The 2-year local, regional, and extrahepatic progression-free survival rates were 78%, 46%, and 39%, respectively. The local control ranked the highest. Patients with Okuda Stage I disease had significantly longer survival than those with Stage II and III (p = 0.02). Patients with T4 disease (p = 0.02) or treated with radiotherapy alone (p = 0.003) had significantly shorter survival. T4 disease (p = 0.03) and pretreatment alpha-fetoprotein level of more than 200 ng/ml (p = 0. 03) were associated with significantly worse regional progression-free survival. A significant difference was observed in both regional progression-free survival (p = 0.0001) and extrahepatic progression-free survival (p = 0.005) between patients with and without portal vein thrombosis before treatment. The presence of satellite nodules had a significantly worse impact on regional progression-free survival (p = 0.04) and extrahepatic progression-free survival (p = 0.03). Patients with hepatic tumor more than 6 cm in diameter or portal vein thrombosis tended to have shorter survival. Radiation-induced liver disease (RILD) and gastrointestinal bleeding were the most common treatment-related toxicities. CONCLUSION: Radiotherapy is effective in the treatment of patients with unresectable HCC. Its effect appeared to be more prominent within the site to which radiation was given. The combination of TACE and radiation was associated with better control of HCC than radiation given alone, probably due to the selection of patients with favorable prognosis for the combined treatment. A dose volume model should be established in the next phase of research in the treatment of unresectable HCC. PMID- 10802372 TI - Socioeconomic characteristics of patients with squamous cell carcinoma of the uterine cervix treated with radiotherapy in the 1992 to 1994 patterns of care study. AB - PURPOSE: To describe the relationship between socioeconomic variables and the treatment of patients with radiotherapy for cervix cancer.40% minority patients. METHODS AND MATERIALS: Sixty-two institutions werw r andomly selected from a list of all radiotherapy facilities in the Unite d States. From these we randomly selected and reviewed 471 cases of squ amous cell carcinoma treated during 1992 to 1994. To create an addition al minority-rich sample, we randomly selected 215 additional cases from 17 randomly selected institutions that admitted >40% minority patients. The median household income of each patient's neighborhood was determined by matching her zip code to data from the 1990 United States Census. RESULTS: Pati ents who lived in low-income neighborhoods, who had only Medicaid covera ge, or who were treated at large academic or minority-rich institutions tended to have a poorer initial performance status, higher-stage or bulk y central disease, and a lower pretreatment hemoglobin level. Ability t o complete treatment did not correlate with ethnicity or income. Howeve r, noncompliant patients tended to be treated at minority-rich instituti ons and were more often < 40 or > 60 years old. Patients who completed definitive treatment were more likely to have had 2 cm (hazard ratio [HR] = 2.66, 95% confidence interval [CI] = 1.16-6.08), and a low biological effective dose (BED) (for each decrease of 5 BED units, HR = 1.76, 95% CI = 1.07-2.88). The 5-year actuarial rate of significant Grade 4 late toxicity was 7.3%. Factors statistically significant for this endpoint on univariate analysis were tumor size (p = 0.035), T-stage (p = 0.02), field size (p = 0.05), fraction size (p = 0.003), and BED (p = 0.05). CONCLUSIONS: RT is an eff ctive treatment option for epithelial skin cancer of the pinna. Large t umor size and low BED were independently statistically significantly ass ociated with increased local failure. Dose-fractionation schedules usin g fraction sizes < 4 Gy may reduce the risk of necrosis and ulceration, particularly for field sizes > 5 cm2. PMID- 10802374 TI - A comparison of two methods for estimating the technical costs of external beam radiation therapy. AB - PURPOSE: To accurately assess the cost-effectiveness of treatment with external beam radiation, it is necessary to have accurate estimates of its cost. One of the most common methods for estimating technical costs has been to convert Medicare charges into costs using Medicare Cost-to-Charge Ratios (CCR). More recently, health care organizations have begun to invest in sophisticated cost accounting systems (CAS) that are capable of providing procedure-specific cost estimates. The purpose of this study was to examine whether these competing approaches result in similar cost estimates for four typical courses of external beam radiation therapy (EBRT). METHODS AND MATERIALS: Technical costs were estimated for the following treatment courses: 1) a palliative "simple" course of 10 fractions using a single field without blocks; 2) a palliative "complex" course of 10 fractions using two opposed fields with custom blocks; 3) a curative course of 30 fractions for breast cancer using tangent fields followed by an electron beam boost; and 4) a curative course of 35 fractions for prostate cancer using CT-planning and a 4-field technique. Costs were estimated using the CCR approach by multiplying the number of units of each procedure billed by its Medicare charge and CCR and then summing these costs. Procedure-specific cost estimates were obtained from a cost-accounting system, and overall costs were then estimated for the CAS approach by multiplying the number of units billed by the appropriate unit cost estimate and then summing these costs. All costs were estimated using data from 1997. The analysis was also repeated using data from another academic institution to estimate their costs using the CCR and CAS methods, as well as the appropriate relative value units (RVUs) and conversion factor from the 1997 Medicare Fee Schedule to estimate Medicare reimbursement for the four treatment courses. RESULTS: The estimated technical costs for the CCR vs. CAS approaches for the four treatment courses were as follows: palliative "simple" $1,285 vs. $1,195; palliative "complex" $2,345 vs. $1,769; curative breast $6,757 vs. $4,850; and curative prostate $9,453 vs. $7,498. Accordingly, the CCR estimates were 8%, 33%, 39%, and 26% higher than the CAS cost estimates, respectively. The primary cause of the difference between the estimates was the daily cost of delivering a "complex" treatment. In fact, if corrected the difference between the estimates fell to 0%, 1%, 4%, and 0%, respectively. Similar results were observed for both methods when the analysis was repeated using data from another academic institution. Medicare reimbursement was also slightly lower than, but remarkably close to, the costs estimated by the CAS approach. CONCLUSIONS: For "complex" treatment courses, which represent the vast majority of external beam treatments, technical costs estimated using the CCR approach appear to be significantly higher than those estimated using procedure specific cost estimates. Because cost-effectiveness analyses of radiation therapy tend to be sensitive to the cost of treatment, the use of higher costs will result in radiation therapy appearing less cost-effective. PMID- 10802375 TI - Association between age and the utilization of radiotherapy in Ontario. AB - PURPOSE: The purpose of this study was to assess whether: (i) radiotherapy (RT) utilization varies with age in Ontario cancer patients; (ii) age-associated differences in the use of RT (if they exist) vary with cancer site and treatment intent; (iii) the age-associated variation in RT utilization is comparable to the decline in functional status in the general population; and (iv) the variation with age is due to differences in referral to a cancer center or to subsequent decisions. METHODS AND MATERIALS: Details for several cancer sites diagnosed between 1984-1994 were obtained from the Ontario Cancer Registry (OCR). RT records from all treatment centers were linked to the OCR database. Information about the functional status of the Canadian population was obtained from the 1994 National Population Health Survey conducted by Statistics Canada. RESULTS: The rate of RT use declined with age, particularly for adjuvant and palliative indications. The relative decline in RT with age exceeded the relative decline in functional status with age in the general population. Most of the decline in RT use was related to a decline in referral to cancer centers. CONCLUSIONS: The referral for, and use of, palliative and adjuvant RT decreases more with age than can be explained by age-associated decline in functional status observed in the general population. PMID- 10802376 TI - MRI measurements correctly predict the relative effects of tumor oxygenating agents on hypoxic fraction in rodent BA1112 tumors. AB - PURPOSE: We evaluate whether magnetic resonance imaging (MRI) with blood oxygenation level-dependent (BOLD) contrast correctly predicts the relative effects of tumor-oxygenating agents on hypoxic fraction in BA1112 rhabdomyosarcomas in WAG/Rij rats. METHODS AND MATERIALS: The response of ten tumors to carbogen (95% O(2)/5% CO(2)), a perfluorocarbon emulsion (PFC), and the combination of PFC + carbogen was studied with high spectral and spatial resolution MR imaging of the water resonance at 4.7 Tesla. Decreases in MR signal linewidth indicate increases in tumor blood oxygen levels. RESULTS: Average MR signal linewidth was decreased 2.0% by carbogen, 2.5% by PFC + air, and 4.9% by PFC + carbogen. PFC + carbogen caused a larger linewidth decrease than either treatment alone (p < 0.04 by ANOVA). Maps of pixels responding to treatment indicate that combining PFC with carbogen significantly enlarges the area of the tumor in which oxygen levels are increased (p < 0.01 by ANOVA). CONCLUSION: MRI predicts that PFC + carbogen will increase radiosensitivity more than either treatment alone; this agrees with the known effects of these treatments on hypoxic fraction. Utilizing MRI to choose the treatment that maximizes the size and extent of increases in tumor oxygenation could reduce hypoxic fraction. PMID- 10802377 TI - The dependence of p53 on the radiation enhancement of thermosensitivity at different let. AB - PURPOSE: The aim of this study is to investigate the dependence of p53-gene status on the radiation enhancement of thermosensitivity at different levels of linear energy transfer (LET). METHODS AND MATERIALS: We used two kinds of human glioblastoma transfectants of A-172 cells bearing the wild-type p53 gene, A 172/neo cells with control vector containing the neo gene and A-172/mp53 cells with both the dominant negative mutated p53 gene and neo gene. We exposed these cells to X-rays and accelerated carbon-ion (C-) beams (13-200 KeV/microm) followed by heating at 44 degrees C. Cellular sensitivities were determined using clonogenic assay. RESULTS: The radiation enhancement of thermosensitivity was LET dependent for the A-172/neo cells, but this was not clearly demonstrated in the A 172/mp53 cells. The supraadditive radiation enhancement of thermosensitivity was observed in A-172/neo cells at the LET range of 13 to 70 KeV/microm, though only an additive effect was observed at higher LET. In A-172/mp53 cells, only an additive effect was observed through all the LET examined. CONCLUSION: These results indicate that the radiation enhancement of thermosensitivity is p53- and LET-dependent. Our results suggest that the combined use of high-LET radiation and hyperthermia brings useful application for cancer therapeutic purposes. PMID- 10802379 TI - Preclinical evaluation of the orally active camptothecin analog, RFS-2000 (9 nitro-20(S)-camptothecin) as a radiation enhancer. AB - PURPOSE: To test for enhancement of radiation effects in vitro and in vivo by the orally administered camptothecin derivative, 9-nitrocamptothecin (RFS-2000); to study whether the mechanism of this enhancement involves inhibition of sublethal damage recovery. METHODS AND MATERIALS: In vitro: H460 human lung carcinoma cells were incubated with RFS-2000 for various times at 37 degrees C, irradiated, immediately rinsed, and assessed for colony-forming ability. Sublethal damage recovery (SLDR) was also assessed using two split doses of radiation. In vivo: H460 cell xenografts were used in nude mice. Tumors were grown subcutaneously on the flank, then treated with RFS-2000 (1 mg/kg) and/or radiation (2 Gy) for 5 consecutive days. Tumor growth delay was then measured for each treatment group. RESULTS: Radiation enhancement was observed in vitro for incubation times between 4 and 24 hr with 10 nM RFS-2000. Using a 24-hr treatment, the radiation dose enhancement ratio values (DER) for 5, 10, and 15 nM were 1.22, 1.54, and 2.0, respectively. Incubation with 10 nM RFS-2000 inhibited SLDR by a factor of 2. The results of three independent in vivo experiments showed that RFS-2000 can enhance the effects of fractionated radiotherapy, with an enhancement factor (EF) of 1.64. CONCLUSION: Our results show that RFS-2000 can enhance the effects of radiation in human lung cancer cells both in vitro and in vivo, and that the mechanism of this effect may involve the inhibition of SLDR. PMID- 10802378 TI - Transfection with mutant p53 gene inhibits heat-induced apoptosis in a head and neck cell line of human squamous cell carcinoma. AB - PURPOSE: To confirm that human cancer cells show p53-dependent heat sensitivity through an apoptosis-related mechanism, we examined the heat sensitivity and Bax mediated apoptosis after heating in a human squamous cell carcinoma cell line, SAS, with identical genetic backgrounds except for the p53 status. MATERIALS AND METHODS: We performed colony formation assay, Western blotting and analyses of apoptosis, using the SAS cells transfected with pC53-248 vector with mutant p53 gene (SAS/Trp248 cells) or the cells transfected with pCMV-Neo-Bam vector (SAS/neo cells) as a control. RESULTS: SAS/Trp248 cells showed heat resistance due to the dominant negative nature of mp53, compared with SAS/neo cells. The incidence of DNA ladders and apoptotic bodies increased markedly after heating in SAS/neo cells, but increased very little in SAS/Trp248 cells. CONCLUSION: These results suggest that heat resistance brought by mp53-transfection is p53 dependent and closely correlates with the induction of apoptosis in human squamous cell carcinomas. PMID- 10802380 TI - Increased tumor cures using combined radiosurgery and BCNU in the treatment of 9l glioma in the rat brain. AB - PURPOSE: Radiosurgery refers to the delivery of high, single focused beams of ionizing radiation to defined intracranial lesions. 1,3 Bis[2-chloroethyl]-1 nitrosourea (BCNU) and cis-diammine-1, 1-cyclobutane-dicarboxylate platinum (II) (carboplatin) are commonly used cytotoxic agents for the treatment of malignant gliomas of the brain. Drug therapies have exhibited a modest enhanced cell killing when combined with radiation in experimental animal tumor systems. The purpose of the present study was to investigate the role of cytotoxic drugs, such as BCNU and carboplatin, in combination with a single high dose of radiosurgery on the tumor control rates of 9L tumors in the rat brain. METHODS AND MATERIALS: Combined radiosurgery (25 Gy single dose) and/or chemotherapy (a single dose of BCNU, 7 mg/kg, i.p. 1.5 or 16 h prior to or 16 h after irradiation or a single dose of carboplatin, 30 mg/kg, administered either 1 h or 4 h prior to irradiation) was delivered 12 days after stereotactic tumor implantation. For dose escalation study, 4-10 mg/kg of BCNU was used. RESULTS: The radiation alone group showed a dose-dependent survival. A single dose of 25 Gy to the control group resulted in an increase of the median survival time from 20 days to 42 days, but all animals died of the tumor in 50 days. A significant prolongation of the median survival time of animals was more than 100 days, resulting in animal cures of 50% or more when combined with radiosurgery (25 Gy) and BCNU (7 mg/kg). BCNU alone did not prolong the median survival time of the animal with the 9L brain tumor. In contrast, there was no survival improvement when the animals were treated with combined radiosurgery and carboplatin. None of the long-term surviving animals showed any significant brain tissue damage as evaluated by histopathology and clinical observations. CONCLUSION: The data clearly suggest that the combined modalities of radiosurgery and concomitant BCNU represent an effective therapeutic regimen in the treatment of radioresistant human malignant gliomas of the brain. This study represents the first experimental report of the effectiveness of combined chemotherapy and radiosurgery. PMID- 10802381 TI - First experience with I-123-alpha-methyl-tyrosine spect in the 3-D radiation treatment planning of brain gliomas. AB - PURPOSE: This study compares the results of iodine-123-alpha-methyl-tyrosine single photon computed emission tomography (IMT-SPECT) with magnetic resonance imaging (MRI) in tumor volume definition of brain gliomas. Furthermore, it evaluates the influences of the information provided from IMT-SPECT for three dimensional (3D) conformal treatment planning. METHODS AND MATERIALS: In 30 patients with nonresected, histologically proven brain gliomas (glioblastoma-13 patients, astrocytoma Grade III-12 patients, astrocytoma Grade II-3 patients, oligodendroglioma Grade III-1 patient, oligodendroglioma Grade II-1 patient), IMT SPECT and MRI were performed pretherapeutically in the same week. A special software system allowed the coregistration of the IMT-SPECT and MRI data. The gross tumor volume (GTV) defined on the IMT-SPECT/T2-MRI fusion images (GTV IMT/T2) was compared with the GTV-T2, defined on the T2-MRI alone. On the IMT SPECT/T1Gd-MRI overlays, the volume of the IMT tumor uptake (GTV-IMT) was compared with the volume of the gadolinium (Gd) enhancement (GTV-T1Gd). The initial planning target volume (PTV) and the boost volume (BV) outlined on the IMT-SPECT/T2-MRI co-images were analyzed comparatively to the PTV and BV delineated using the T2-MRI alone. RESULTS: In all 30 patients a higher IMT uptake of tumor areas, compared to the normal brain tissue was observed. Mean GTV IMT, mean GTV-T2, and mean GTV-T1Gd were 43, 82, and 16 cm(3), respectively. IMT tumor uptake outside the contrast enhancement regions was observed in all patients. Mean relative increase of tumor volume defined on the fusion images, GTV-IMT/T1Gd versus GTV-T1Gd alone was 78%. IMT tumor uptake areas outside the GTV-T2 were registered in 7 patients (23%). In these patients, the mean increase GTV-IMT/T2 was 33% higher than GTV-T2, defined according to the T2-MRI data alone. The additional information provided by IMT-SPECT modified minimally the initial PTV (mean relative increase PTV-IMT/T2 versus PTV-T2, 5%) but significantly the BV (mean relative increase BV-IMT/T2 versus BV-T2, 37%). CONCLUSION: In a significant number of patients, the IMT-SPECT investigation improves tumor detection and delineation in the planning process. This has important consequences in the 3D conformal treatment planning, especially in the delineation of BV and in dose escalation studies. PMID- 10802382 TI - Design and dosimetry characteristics of a soft-docking system for intraoperative radiation therapy. AB - PURPOSE: The design concept and the dosimetric characteristics of an applicator system for intraoperative radiation therapy (IORT) with special emphasis on alignment methods, the effect of a plastic scatterer in the beam, radiation leakage, and misalignment dosimetry, are presented in this paper. MATERIALS AND METHODS: A soft-docking system for a linear accelerator, which enables collimation of electron beams (4-22 MeV) for IORT has been developed. The system includes twenty-one circular polymethylmethacrylate (PMMA) treatment cones of different lengths, diameters and end angles. All in-water measurements are made using p-type silicon diode detectors. RESULTS: The effect of introducing a PMMA scatterer in the therapeutic beam includes increased surface dose values (above 83% for all nominal electron energies and for all cones) and improved dose homogeneity within the therapeutic range. Electrons scattered from the inside wall of the cone result in dose profile horns at depth of dose maximum always lower than 109%. The radiation leakage outside the cone is less than 13%. Large changes in the dose profiles occur if the intraoperative cone is misaligned more than 0.5. CONCLUSION: The alignment procedure of the soft-docking system is easy to handle and the applicator design provides adequate collimation of electron beams for IORT. PMID- 10802384 TI - Contribution of protein kinase C and glutamate in Pb(2+)-induced cytotoxicity. AB - Activation of protein kinase C (PKC) plays an important role in lead (Pb(2+)) induced cytotoxicity. The effects of low dose exposure to Pb(2+) on cytosolic free calcium (Ca(2+)), PKC activity and mechanisms involved in cell death were studied in PC12 cells. Exposure of PC12 cells to low dose Pb(2+) (0.01 microM) increased PKC activity, while exposure to a higher dose (10 microM) led to decreased PKC activity. Additionally, in normal extracellular medium, low concentration of Pb(2+) (0.01 microM) stimulated increase in cytosolic free calcium while the higher concentrations of Pb(2+) (10 microM) did not. However, the effect of low dose Pb(2+) (0.01 microM) was blocked by removing Ca(2+) from external medium. The role of Pb(2+)-induced changes in PKC activity and its relationship to oxidative stress and related cytotoxicity was also studied. Pb(2+) alone (0.01-10 microM) produced reactive oxygen species (ROS) dose dependently over the period of 24 h. Pb(2+)-induced ROS were potentiated in the presence of 500 microM glutamate. Furthermore, a correlation was observed between ROS generation and the levels of cytotoxicity, which was observed after 24 h exposures to Pb(2+) by trypan blue method, and the cytotoxicity was enhanced by glutamate co-treatment. Pb(2+)-induced cell death was blocked partially by staurosporine (PKC inhibitor, 100 nM) and NMDA antagonist, MK-801 (1 microM). It is concluded that, in Pb-induced cytotoxicity, modulation of PKC and intracellular calcium play significant roles in augmenting glutamate receptor mediated oxidative species formation and subsequent cell death. PMID- 10802385 TI - Induction of apoptosis in mouse thymocytes by cadmium. AB - In the thymus apoptosis is an important process in T cell maturation and differentiation. Cadmium (Cd) is an ubiquitous toxic metal that is capable of modulating immune responses. To investigate the induction of apoptosis and immunomodulation by environmental chemicals, we cultured mouse thymocytes with Cd and/or dexamethasone (DEX). DNA fragmentation was analyzed by gel electrophoresis, ELISA and flow cytometry. Treatment with either Cd or DEX induced DNA fragmentation in the thymocytes. Exposure to 10 microM Cd killed thymocytes by apoptosis rather than necrosis. However, no synergistic or additive effect was observed in the induction of apoptosis when DEX was added to the Cd. These results suggest that Cd may modulate the function of the thymus by the induction of apoptosis through mechanisms that differ from those used by DEX. PMID- 10802386 TI - Induction of class alpha glutathione S-transferases by 4-methylthiazole in the rat liver: role of oxidative stress. AB - The expression of glutathione S-transferase (GST) is a crucial factor in determining the sensitivity of cells and organs in response to a variety of toxicants. Expression of class alpha GST genes by methyl-substituted thiazoles was assessed in the rat liver. Northern blot analysis revealed that 4 methylthiazole (4-MT) elevated rGSTA2, A3, A5 and M1 mRNAs in the liver by 19-, 4 , 6- and 9-fold at 24 h after treatment, respectively, as compared to control. Consecutive 3-day treatment with 4-MT resulted in 4- to 7-fold increases in rGSTA and M1 mRNAs. Multiple treatments with 5-methylthiazole (5-MT) caused marginal increases in GST mRNAs in spite of the large increases in certain GST mRNAs at 24 h. Either 4, 5-dimethylthiazole (DT) or 2,4,5-trimethylthiazole (TT) minimally affected the rGSTA and rGSTM mRNA expression at 1-3 day(s). Western blot analysis showed that 4-MT induced rGSTA1/2, rGSTA3/5 and rGSTM1 proteins by 2.6-, 2.1- and 2.1-fold at 3 days, respectively, while other methylthiazoles failed to induce the GST subunits. Starving rats were treated with a lower dose of methylthiazoles to study the role of oxidative stress in the mRNA expression. The levels in rGSTA2/3/5 mRNAs were significantly enhanced by 4-MT in starving rats, whereas rGSTM1/2 mRNAs were not further increased. Other methylthiazoles were inactive in enhancing the mRNAs in starving animals. Pretreatment of starving rats with either cysteine or methionine completely prevented the increases in class alpha GST mRNAs by 4-MT. Data showed that 4-MT induces class alpha GSTs with the increases in the mRNAs, whereas 5-methyl-, dimethyl- and trimethyl-substituted thiazoles were minimally active. Increases in the class alpha GST mRNAs by 4-MT may be associated with the oxidative stress in hepatocytes, as supported by starvation and sulfur amino acid experiments. PMID- 10802387 TI - Glutamate and aspartate impair memory retention and damage hypothalamic neurons in adult mice. AB - We examined the effects of systemic administration of monosodium glutamate (MSG) or aspartate (ASP) on the memory retention and neuronal damage in the brains of adult mice. Compared with the control mice, a single intraperitoneal injection of either 4.0 mg/g MSG or 0.5 mg/g ASP after acquisition trial significantly shortened the response latency in the passive avoidance test, accompanying by the transient weight loss. Histopathological analysis of the brains of these mice revealed that neurons in the arcuate nucleus of hypothalamus were damaged markedly by MSG (4.0 mg/g) or ASP (0.5 mg/g). Other brain areas including cerebral cortex and hippocampus did not show any pathological changes. These findings suggest that systemic administration of MSG or ASP could impair memory retention and damage hypothalamic neurons in adult mice. PMID- 10802388 TI - Expression of heat shock protein 60 in human proximal tubule cells exposed to heat, sodium arsenite and CdCl(2). AB - The expression of hsp 60 mRNA and protein were determined in human proximal tubule cells (HPT) exposed to lethal and sub-lethal concentrations of Cd(2+) under both acute and extended conditions of exposure. It was demonstrated that HPT cells exhibited the classic heat shock response when subjected to a physical (heat) or chemical stress (sodium arsenite). Heat stress, elevated temperature at 42.5 degrees C for 1 h, caused an increase in both hsp 60 mRNA and protein following removal of the stress. Similar results were obtained when the cells were subjected to a classic chemical stress of exposure to 100 microM sodium arsenite for 4 h. Acute exposure of HPT cells to 53.4 microM CdCl(2) for 4 h also resulted in an increase in hsp 60 mRNA and protein following removal of the metal. An extended exposure to Cd(2+) was modeled by treating the cells continuously with Cd(2+) at both lethal and sub-lethal levels over a 16-day time course. It was demonstrated that chronic exposure to Cd(2+) failed to increase either hsp 60 mRNA or protein expression in HPT cells, even at concentrations of Cd(2+) that were lethal to the cells during the time course. In fact, hsp 60 protein levels were decreased compared to controls at lethal levels of Cd(2+) exposure. These findings suggest that hsp 60 expression may have two distinct roles when the human proximal tubule cell is exposed to Cd(2+). A protective role through hsp 60 induction when the proximal tubule cell is acutely exposed to Cd(2+) and a deleterious role when hsp 60 protein is down-regulated during extended exposure to Cd(2+). PMID- 10802389 TI - Calcium associated resistance to H(2)O(2) in Chinese hamster V79 cells. AB - To investigate whether the difference in cellular sensitivity of Chinese hamster V79 and their H(2)O(2)-resistant variant cells (Hpr-4) to H(2)O(2) relates to the difference in intracellular Ca(2+) concentration in these cells, we measured Ca(2+) concentration by calcium ion analysis after loading these cells with Fura 2/AM. Intracellular Ca(2+) concentration increased in both Chinese hamster V79 and Hpr-4 cells as extracellular Ca(2+) concentration increased. However, the increase in intracellular Ca(2+) concentration in response to extracellular H(2)O(2) was more pronounced in Hpr-4 than V79 cells. H(2)O(2) cytotoxicity of Hpr-4 but not V79 cells was also decreased in response to the increase in extracellular Ca(2+) concentration. In parallel with the decrease in cytotoxicity in response to increasing extracellular Ca(2+) concentration, the frequency of mitochondrial DNA single strand breaks (SSB) in Hpr-4 cells also decreased without producing observable nuclear DNA SSB. Use of permeabilized V79 and Hpr-4 cells exposed to H(2)O(2) showed that mitochondrial DNA SSB decreased when extramitochondrial Ca(2+) concentration increased. These findings indicate that elevated intracellular Ca(2+) concentration may protect against H(2)O(2)-induced mitochondrial damage and cytotoxicity in these cells. PMID- 10802390 TI - The effects of n-alcohols on evoked synaptic potentials in rat hippocampal slices: Hill coefficients account for the cut-off phenomenon. AB - The anesthetic potencies of n-alcohols increase progressively with lengthening of the carbon chain and then disappear at a cut-off point of a longer-chain n alcohol. In order to assess the mechanisms for cut-off in mammalian central nervous system, the effects of a series of n-alcohols (C(2)-C(11)) were examined on the evoked synaptic potentials of the rat hippocampal preparation in vitro. The n-alcohols (C(2)-C(10)) reduced the slope of the excitatory post-synaptic potential in a concentration-dependent manner, and the inhibitory potencies enhanced as a function of carbon chain length. The effect disappeared at n undecanol (C(11)). The Hill coefficients of the concentration-response curves of the n-alcohols negatively correlated with the number of carbon atoms. The decrease in the Hill coefficient could account for the cut-off phenomenon, indicating that the results can support the anesthetic pocket hypothesis. PMID- 10802391 TI - Depletion of cellular glutathione by conditions used for the passaging of adherent cultured cells. AB - Cultured cells are commonly exposed to trypsin-containing solutions in order to prepare cell suspensions suitable for subculture. Conditions used to release and disperse monolayers of cultured murine hepatoma 1c1c7 and human breast epithelial MCF10A cells caused the loss (40-95%) of cellular glutathione (GSH), but did not affect viability. Glutathione contents returned to pretrypsinization values within 24 h of replating. In contrast, the GSH contents of cultured rat hepatoma 5L cells were not affected by trypsinization. Exposure of 1c1c7 cultures to H(2)O(2) or etoposide 1 or 24 h after replating resulted in concentration dependent cytostatic and cytotoxic effects. The concentration-response curves defining the cytostatic and cytotoxic effects of etoposide, and the cytostatic effects of H(2)O(2) were not influenced by the timing of toxicant addition. However, 1c1c7 cultures treated with H(2)O(2) 1 h after replating were more susceptible to the cytotoxic actions of the peroxide than cultures treated 24 h after plating. These studies show that conditions commonly used for the passaging of cultured cells can lead to a transient, but profound loss of GSH in some cell lines. Furthermore, the outcome of cytotoxicity analyses can be influenced by the time elapsed between the plating of cultures and the addition of toxicant. PMID- 10802392 TI - Cytotoxicity and keratinocyte microsome-mediated mutagenic activation of carcinogens in cultured epidermal cells. AB - Four model carcinogens (aflatoxin B(1), 6-nitrochrysene, 3-amino-1-dimethyl-5H pyrido[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2)) were examined for their ability to inhibit the growth of cultured human and rat epidermal cells. To find a basis for observed differences in growth inhibition, aflatoxin B(1), Trp-P-1 and Trp-P-2 were tested for activation by microsomes isolated from these cells in a bacterial mutagenesis assay. Treated rat cultures exhibited sensitivity to Trp-P-1 and Trp-P-2 and especially aflatoxin toxicity (growth inhibition) despite their microsomes being unable to induce bacterial mutagenicity. In treated human cultures, the toxicities of Trp-P-1, Trp-P-2 and AFB(1) were stimulated by 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD), consistent with their dependence on the biotransformation reactions this agent induces; however, the toxicity correlated poorly with observed bacterial mutagenicity mediated by their isolated microsomes. 6-Nitrochrysene, a known direct-acting mutagen in bacteria, was highly toxic to the rat but not to the human cells. Since toxic effects can modify carcinogenic outcomes, these findings are compatible with a complex relationship between toxicity, mutagenicity and carcinogenicity and indicate the utility of keratinocytes for clarifying this relationship. PMID- 10802393 TI - Maturitas, the european menopause journal, becomes 'Official journal of the european menopause and andropause society' PMID- 10802394 TI - Factors associated with early menopause. AB - OBJECTIVE: An understanding of why certain factors contribute to a more rapid decline in ovarian function may, for some women, help prevent premature loss of fecundity and the subsequent impact of health problems secondary to long-term estrogen deficiency such as osteoporosis, cardiovascular disease, and possibly Alzheimer's disease. METHODS: A summary of the evidence regarding factors that have been proposed to contribute to an early onset of natural menopause is presented. These factors include cigarette smoking, race, education, parity, menstrual cycle length, the use of oral contraceptives, age at menarche, major depression, anthropometry, and handedness. RESULTS: Cigarette smoking has been found to hasten the onset of menopause by as much as one year. Lifetime number of ovulatory cycles (indicative of oocyte depletion) is also predictive of the age at natural menopause (ANP). This is consistent with the many studies that have reported early ANP among women with shorter menstrual cycles, and a later ANP among multigravid women or those who used oral contraceptives. The relationship between depressive disorder and ovarian failure is complex, involving consideration of the pharmacological effects of treatment, and is currently unclear. The findings regarding an effect of body mass index on ANP are also mixed. At this time, there is little persuasive evidence that handedness or demographic characteristics (independent of their relationship with behavioral factors like smoking) influence the ANP to any substantial degree. CONCLUSIONS: Some factors that could potentially influence ANP have been identified, but these and other avenues of investigation warrant further study. PMID- 10802395 TI - Conceptualizing menopause and midlife: Chinese American and Chinese women in the US. AB - OBJECTIVES: The purpose of this qualitative project was to describe and examine expectations and experiences of menopause and midlife in pre- and postmenopausal Chinese American and Chinese women in the United States. METHODS: Four focus groups were formed from a total of 44 women: two groups of premenopausal Chinese/Chinese American women (one conducted in English and one in Cantonese) and two groups of postmenopausal Chinese/Chinese American women (one conducted in English and one in Cantonese). Qualitative data, in the form of transcripts, were interpreted using text-based content analysis. RESULTS: The major thematic findings were: meanings of menopause are inextricably bound with meanings of midlife; the borders and timing of the menopausal transition are ambiguous; the menopausal transition represents a natural progression through the life cycle; the expectations of the premenopausal women did not match the experiences of the postmenopausal women; menopause is viewed as a marker for aging; and the menopausal transition must be prepared for and managed. CONCLUSIONS: Study findings indicate that the participants did not share the strictly medicalized view of menopause as a discrete, biological entity. Menopause was typically described as a natural transition that was virtually interchangeable with midlife. While most of the participants characterized menopause as signaling the end of fertility and virtually synonymous with old age, some women described it as a new opportunity and a second chance at life. Participants felt a sense of their own agency in addressing what they viewed as a complex life stage, the experience of which could be manipulated. PMID- 10802396 TI - The effects of physical exercise on body fat distribution and bone mineral density in postmenopausal women. AB - OBJECTIVE: The present cross-sectional study investigated the effects of physical exercise on body fat distribution and bone mineral density (BMD). METHODS: Subjects were 57 postmenopausal women (mean age, 60.5+/-6.4 years) who had exercised regularly for at least 2 years. Controls were 130 age-matched sedentary women. Age, years since menopause (YSM), height, weight, and body mass index (BMI, wt./ht.(2)) were recorded. Total fat mass, percentage of body fat, trunk fat mass, leg fat mass, the ratio of trunk fat mass to leg fat mass (trunk-leg fat ratio), total body lean mass, percentage of body lean, and lumbar spine BMD (L2-L4) were measured by dual-energy X-ray absorptiometry. RESULTS: Baseline characteristics and leg fat mass did not differ between the two groups. Total fat mass, percentage of body fat, trunk fat mass, and trunk-leg fat ratio were lower (P<0.05, P<0.01, P<0.01 and P<0.001, respectively), while total body lean mass, percentage of body lean mass, and lumbar spine BMD were higher in exercising women (P<0.05, P<0.05 and P<0.01, respectively). Performing physical exercise was inversely correlated with trunk-leg fat ratio (standardized regression coefficient=-0.178, P<0.01), but positively correlated with BMD (0. 203, P<0.01) irrespective of age, height, YSM, and total fat mass. CONCLUSION: Physical exercise has beneficial effects on body fat distribution and BMD in postmenopausal women. Reduction of upper body fat distribution with physical exercise may be more attributable to the decrease in trunk fat mass. PMID- 10802397 TI - A comparison of endothelial function in Caucasian and Chinese women before and after the menopause. AB - OBJECTIVE: Arterial endothelial dysfunction is a key atherogenic event that may be related to oestrogen status. We therefore aimed to compare menopause-related changes in endothelial physiology in Chinese and Caucasian females. METHODS: We studied 40 female subjects; 20 Chinese from a rural region of Southern China (ten premenopausal, aged 20-35 years, and ten postmenopausal, aged 55-66 years), and 20 age-matched Caucasian females from Sydney, Australia. All women had a clinical history, resting blood pressure and fasting lipids measured, and endothelial function assessed. Using high-resolution external vascular ultrasound, brachial artery diameter was measured at rest, after flow increase (causing endothelium dependent dilatation) and after sublingual glyceryl trinitrate (GTW) (an endothelium-independent dilator). RESULTS: There was a significant decline in endothelium-dependent dilatation (EDD) comparing Caucasian females from the premenopausal versus postmenopausal groups (8.4+/-2.7% versus 2.7+/-2.9%; P<0.001). In contrast, there was no significant difference in EDD between pre- and postmenopausal Chinese (9.8+/-3.3% versus 8.3+/-1.7%; P=0.22). On multivariate analysis, postmenopausal status was associated with impaired EDD in Caucasian females (P<0.002) independent of serum cholesterol, blood pressure and vessel size. In contrast, EDD in the Chinese females was not significantly influenced by any of these factors. GTW-induced dilatation was also impaired in the older Caucasian females, a finding explained on multivariate analysis by the increased vessel size in this patient group (P=0.03). CONCLUSION: Menopause is associated with impaired arterial endothelial function in Caucasian but not Chinese women. This suggests possible ethnic differences in menopause-related vascular changes. PMID- 10802398 TI - Behaviour of the carotid wall in menopausal women with and without arterial hypertension. AB - OBJECTIVE: A study was conducted on the carotid vascular wall to evaluate its behaviour in postmenopausal women suffering from arterial hypertension compared to normotensive postmenopausal women. METHOD: Intima-media thickness(IMT) of carotid artery was determined by ecoduplex scanner in 182 postmenopausal women (age range 40-60 years) divided in four age classes (40-45, 46-50, 51-55, 55-60 years). Ninety-one women presented normal arterial blood pressure (SBP 136.6+/ 11.7 and DBP 82.3+/-8.9 mmHg) and 91 were hypertensive (SBP 172.6+/-11.7 and DBP 97.4+/-6.6 mmHg). RESULTS: The normotensive women presented the following IMT values: 1.19+/-0.21 mm (40-45 years), 1.21+/-0.25 mm (46-50 years), 1.25+/-0.20 mm (51-55 years), 1.25+/-0.20 mm (55-60 years). IMT values in hypertensive women were: 1.75+/-0.25 mm (40-45 years), 1.77+/-0.30 mm (45-50 years), 1.91+/-0.28 mm (51-55 years), 2.02+/-0.33 mm (55-60 years). ANOVA test was performed in both groups of women and did not show any significant difference in the four age classes of normotensive women. On the other hand it revealed a statistical significance between 40-45 and 46-50-year-old hypertensive classes (P<0.001) and between 51-55 and 55-60-year-old hypertensive clases (P<0.02). CONCLUSIONS: There was a correlation between age and IMT in the arterial hypertensives, especially in two older classes, and between the duration of menopause and IMT values. The results indicated that carotid wall thickening was constantly higher in hypertensive women compared with normotensive ones. PMID- 10802399 TI - Acute effects of estradiol and progesterone on insulin, lipids and lipoproteins in postmenopausal women: a pilot study. AB - OBJECTIVES: To examine the acute effects of estradiol-17beta (E(2)) and progesterone (P) on serum levels of insulin, lipids and lipoproteins in estrogen deficient postmenopausal women, whereby, a direct cause-effect relationship could be established without the influence of lifestyle changes. MATERIALS AND METHODS: Nine postmenopausal women were given oral E(2) (Estrace) 2 mg/day for 28 days and oral micronized P (Prometrium) 200 mg/day in the last 14 days of E(2) treatment. Fasting blood samples were obtained before starting E(2) (day 1) and P (day 15) and on day 29. Serum levels of insulin, triglycerides (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL) and lipoprotein (a) (Lp(a)) at the three time points were compared by Friedman analysis of variance (ANOVA). Corresponding levels of glucose, the apolipoproteins (Apo) A1 and B and serum androgen levels were also evaluated. RESULTS: E(2) decreased while P increased fasting levels of insulin (32.45+/ 3.57, 26.36+/-2.90 and 37.36+/-3.67 pmol/l on day 1, 15 and 29 respectively; P<0.01). Fasting glucose to insulin ratios changed inversely (P<0.01). E(2) increased HDL from 1.07+/-0.05 mmol/l on day 1 to 1.17+/-0.07 mmol/l on day 29 but decreased corresponding levels of Lp(a) from 261+/-93 to 211+/-83 U/l (P=0.03 for both). TC and LDL levels fell significantly after 14 days of E(2) treatment with no further decrease when P was added. Androgen levels remained unchanged during hormone treatment. CONCLUSION: The sequential, acute effects of E(2) and micronized P on insulin and lipids confirm a direct cause-effect relationship. The acute effects of P on insulin in particular, highlights the importance of standardizing the medication days according to estrogen and progestin in the clinical evaluation of their true metabolic impact in longer-term studies and may influence the choice of progestin type, dose and duration in hormone replacement. PMID- 10802400 TI - Effect of dose on the absorption of estradiol from a transdermal gel. AB - OBJECTIVES: To study whether dose adjustments in transdermal estradiol gel treatment would result in proportional changes in estradiol bioavailability and concentrations. METHODS: In an open study, 23 healthy postmenopausal women were treated consecutively with 0.5, 1.0 and 1.5 mg estradiol daily as a transdermal gel. Each dose was given for 16 days. Venous blood samples for serum estradiol and estrone measurements with RIA were taken at steady state on the 16th study day. From these concentrations, pharmacokinetic parameters for estradiol were calculated and corrected to correspond to equal dose by dividing the values by the dose. RESULTS: Area under the estradiol time-concentration curve and peak estradiol level increased linearily and dose-proportionally with daily estradiol doses of 0.5-1.5 mg. This was shown by lack of significant differences in the dose-corrected parameters. However, the 90% confidence intervals between the doses were outside the commonly accepted levels for bioequivalence. Peak estradiol level was clearer and occurred earlier with the highest 1.5 mg estradiol dose, while more stable estradiol levels were seen with the lowest 0.5 mg estradiol dose. CONCLUSIONS: The amount of estradiol on a certain skin area seems to be the determining factor in absorption. With higher estradiol doses, the absorption will be accelerated with a clearer peak estradiol level. The linear and dose proportional absorption indicates that flexible dose adjustments within the dose range of 0.5-1.5 mg estradiol daily can be made with an anticipated effect in estradiol bioavailability and concentrations. PMID- 10802401 TI - Application and maintenance habits do make a difference in adhesion of Alora transdermal systems. AB - OBJECTIVES: To explore and evaluate Alora placebo patch application and maintenance habits of women in order to identify the factors that influence adhesion success. METHODS: This single-center, open-label, placebo, randomized, multiple-application, parallel-group study involved 99 healthy naive users of transdermal patches. Participants applied and wore an Alora placebo patch for ten consecutive applications of approximately 3.5 days each and evaluated adhesion of the patches twice-daily. Three subgroups comprising participants achieving low, moderate or high adhesion success took part in focus groups to discuss their wear habits, practices and attitudes regarding transdermal patches. RESULTS: There was a significant behavioral component involved in patch adhesion. The habits, practices and attitudes of high achievers were clearly different from the other two subgroups. The three most important issues identified to improve adhesion were: mastering the removal of the patch liner, identifying the best site of application, and developing and implementing techniques to maintain patch adhesion. The Alora placebo patch was well tolerated throughout the study. CONCLUSION: The data showed that there is a learning curve involved in achieving maximal adhesion with a transdermal patch. During the study, a novel patch application method ('press, fold and slide') was demonstrated for the participants. This method was very well received by all participants and was more easily executed than the previous method. An adaptation of this method was incorporated into the Alora patient information leaflet, together with several other changes to help improve adhesion success. PMID- 10802402 TI - Hormone replacement therapy in women with pre-existing hyperpigmented lesions. AB - OBJECTIVES: This study was performed to determine whether skin pigmentation is darkened after hormone replacement therapy (HRT). METHODS: The color of hyperpigmentary lesions and control sites before and after 1, 2, and 3 months of HRT was measured. RESULTS: All of the three tested sites showed no significant pigment alteration in 3 months of follow up after starting HRT (P>0.05). Age, duration of menopause, and sex hormone levels did not correlate with pigmentation level. CONCLUSIONS: Pigmentation changes after HRT are not significantly associated with the treatment. This finding suggests that low-dose estrogen replacement therapy does not induce pigmentation changes alone and that differences in individual susceptibility and end organ responsiveness or other multiple factors in addition to the sex hormone may be responsible for the development and darkening of hyperpigmentary lesions. PMID- 10802403 TI - Self-rated health, life satisfaction and personal characteristics of post menopausal women under estrogen replacement therapy. AB - OBJECTIVES: To describe the prevalence of oestrogen replacement therapy (ERT)-use among post-menopausal women in relation with personal and socio-economic characteristics and to examine the association of ERT-use with self-rated health and selected aspects of life satisfaction. METHODS: Population survey data were derived from a cohort study of 511 Bernese women, aged 55-65 years. Data were collected by means of telephone interviews. Overall prevalence of ERT-use, and selected associations with personal and socio-economic characteristics were investigated using descriptive statistical methods and logistic regression. The relations of ERT-use with five self-reported health measures were explored using Spearman's correlation coefficients. The associations of ERT-use with six dichotomous variables on satisfaction with various aspects of life were tested with chi-square tests in cross tabulations. RESULTS: The overall prevalence of self reported ERT-use was 17.6%. ERT was more prevalent women younger than 61 years than in women in the older age group. Women with a body mass index (BMI) under 25 reported ERT-use significantly more frequently than women with higher BMI (OR=3.16, CI 1.87-5.34). ERT-use was more prevalent in women with relatively high education: OR=2.01, CI 1.18-4.00. The self-reported health measures and the satisfaction items were not significantly associated with ERT-use. CONCLUSIONS: ERT-use among post-menopausal women was found to be associated with higher educational level and lower BMI. ERT-users did not report better health or life satisfaction. PMID- 10802404 TI - Absent correlation between vaginal bleeding and oestradiol levels or endometrial morphology during tibolone use in early postmenopausal women. AB - OBJECTIVE: To investigate a potential correlation between vaginal bleeding and oestradiol (E2) levels/endometrial morphology in early postmenopausal women using tibolone (Livial(R)). METHODS: A 2-year randomised placebo-controlled study of 94 healthy women, 1-3 years after spontaneous menopause, receiving either placebo (n=23), 1.25 mg/day (n=36) or 2.5 mg/day (n=35) tibolone. Episodes of vaginal bleeding throughout the 2-year study period were recorded. Age, age of menopause, months since menopause and body mass index were recorded. Serum E2 levels were assessed at baseline and at 3-month intervals throughout the study period. In case of vaginal bleeding, endometrium morphology was assessed by Vabra Curettage. RESULTS: Fifty-one percent (n=18, P<0.05) of women in the 2.5 mg/day tibolone group and 44% (n=16, P=0.07) in the 1.25 mg/day tibolone group presented with at least one period of vaginal bleeding, compared with 22% (n=5) in the placebo group. The women who bled in the placebo group were younger (P<0.01), had menopause at an earlier age (P<0.05), had a shorter duration since menopause (P<0.05) and had a higher median E2 serum level prior to bleeding (P<0.05). In contrast, in both tibolone groups, no determinants could be found for the vaginal bleeding. Ninety percent of the first bleedings occurred within 9 months after starting the treatment. At Vabra endometrium sampling, there was no evidence of endometrial stimulation. CONCLUSIONS: In the present study, early postmenopausal women using 1.25 or 2.5 mg/day tibolone are 2-2.5 times more likely to present with vaginal bleeding compared with placebo (P<0.05) without evidence of higher serum E2 levels or endometrial stimulation. PMID- 10802405 TI - Formulation and evaluation of ophthalmic preparations of acetazolamide. AB - The orally administered acetazolamide has a limited use in glaucoma due to the systemic side effects associated with its use. It has been reported to show little effect on the intraocular pressure (IOP) of human and rabbit eyes upon topical application, probably owing to its poor bioavailability and instability at pH >5.0. In order to enhance the bioavailability of the drug, contact time between the drug molecules and the ocular surface was increased using high viscosity, water soluble polymers (PVA, HPMC), and by incorporating acetazolamide into an in situ-forming ophthalmic drug delivery system. Moreover, a penetration enhancer (EDTA) was also used in these formulations to increase the extent of absorption of the drug. Acetazolamide at a concentration of 10% was used and the formulations (eyedrop suspensions) were evaluated for their in vitro release pattern. The effect of these formulations on the IOP in normotensive conscious rabbits was also investigated. These formulations were found to be therapeutically effective with a peak effect at 2 h. A fall in IOP of up to 46.4% was observed with repeated administration of one of the formulation containing PVA, EDTA and Tween 80 (MK-5). Results indicated that a topical effect of acetazolamide can be observed if the formulation, (a) contains a suitable polymer to increase the residence time; (b) a penetration enhancer-as acetazolamide has a low permeability coefficient i.e. 4. 1x10(-6) cm/s [Duffel, M.W., Ing. I.S., Segarra, T.M., Dixson, J.A., Barfknecht, C.F., Schoenwald, R.D., 1986. J. Med. Chem. 29, 1488-1494]; and (c) pH of the formulation is maintained at the point of maximum stability (pH< or =5.0). PMID- 10802406 TI - Formulation and process considerations for beads containing Carbopol 974P, NF resin made by extrusion-spheronization. AB - Preliminary studies revealed that Carbopol 974P, NF resin could be incorporated into beads manufactured by extrusion and spheronization, and can slow the release of a highly water soluble drug if calcium chloride was included in the granulating fluid to reduce the tack of the wetted polymer. In this study, the same approach was used to produce high quality chlorpheniramine maleate beads with a prolonged release duration. Because of the complex nature of the extrusion and spheronization process and the various components in the bead formulations, a statistically sound factorial experiment was considered for this study. A one half fraction of a two level factorial design with three center points was employed to estimate the effects of simultaneously modifying multiple process and formulation variables, including the Carbopol concentration, calcium chloride concentration, water content, and the spheronization speed and time. Product yield, average bead roundness, and the drug release profile were selected as responses. Increasing the Carbopol content across the experimental range resulted in a significant (P<0.05) reduction in the percentage drug released at 25, 40, and 60 min. Results suggest that combining the conditions of high Carbopol, high water, and low calcium chloride levels with low spheronization speeds at long spheronization times produce the highest quality bead with the longest drug release duration. PMID- 10802407 TI - An investigation into the use of stepwise isothermal high sensitivity DSC as a means of detecting drug-excipient incompatibility. AB - The use of stepwise isothermal high sensitivity differential scanning calorimetry (HSDSC) as a novel means of detecting excipient incompatibility is described using aspirin mixes with magnesium stearate and stearic acid as model systems. Aspirin, magnesium stearate and stearic acid alone and as mixes were studied in scanning mode using conventional DSC and were then subjected to a stepwise heating programme using HSDSC, whereby the samples were heated to temperatures between 45 and 70 degrees C and held for 1 h, during which the heat flow to or from the sample was measured. The data indicated that while no thermal events were detected for the individual components or mixes with stearic acid other than melting of stearic acid, 50% w/w mixes of magnesium stearate showed a marked endothermic response at temperatures above 55 degrees C. The data were fitted to an adaptation of an existing kinetic model for the degradation process and a reasonable correlation found. Mixes of the drug with the two excipients were then studied at 60 degrees C over 6 h at concentrations between 1 and 50% w/w. Incompatibilities with magnesium stearate concentrations as low as 1% w/w could be detected using this approach. Compacts of magnesium stearate and aspirin were also studied, with considerably more pronounced thermal events taking place compared to the powder mixes. It is concluded from these studies that while the study has highlighted certain limitations of the approach, stepwise isothermal DSC represents a potentially highly useful means of detecting excipient incompatibilities. PMID- 10802408 TI - Controlled-release liquid suspensions based on ion-exchange particles entrapped within acrylic microcapsules. AB - Eudragit RS/RL polymers were used to prepare microcapsules containing terbutaline loaded ion-exchange resins, with the final aim of formulating this anti-asthmatic drug in a controlled-release liquid form. Oil-in-oil (o/o) and oil-in-water (o/w) solvent evaporation procedures were conveniently modified in order to encapsulate the resin cores. The microcapsules were then suspended in a hydroxypropylmethylcellulose solution of adequate viscosity and palatability, and stored at 20 degrees C and ambient humidity conditions for a 6-month period. Stability studies of the dispersed microparticles were performed in order to evaluate the changes occurred in the diffusion of the drug to the suspending medium and in the dissolution behaviour during storage. The morphological alterations of the stored microcapsules were followed throughout the duration of the study by scanning electron microscopy. The polymer coatings of microcapsules prepared by the o/o method broke up on the first day of storage, while those made by the aqueous procedure remained intact during all the storage period. This agreed with the modification observed in the controlled-release profiles of terbutaline in the case of microcapsules prepared by the o/o method, which completely changed after the first week of storage. On the contrary, the microcapsules prepared by the aqueous method showed identical controlled-release profiles for all the stability study. The different behaviour of both types of microcapsules was attributed to the swelling suffered by the resin particles in contact with the aqueous suspending medium, which was higher in the microcapsules prepared by the o/o technique. In fact, in the anhydrous procedure, the microencapsulation was carried out on the shrunken resin particles, whereas in the o/w method, the presence of water during the microencapsulation process allowed the coating of the swollen particles, thus avoiding the further problem of rupture of the polymer coating. PMID- 10802409 TI - Stereoselective pharmacokinetics of ibuprofen and its lysinate from suppositories in rabbits. AB - Studies were performed on the effect of ibuprofen racemate ionisation extent on the pharmacokinetics of its enantiomers following administration in suppositories to rabbits. The suppositories, containing 146.3 mg ibuprofen in acidic form (IBP) or 250 mg ibuprofen lysinate (IBPL), equivalent to the above IBP dose, were prepared using lipophilic Witepsol H-15 as a base and administered to rabbits in a crossover design. Compared with IBP, administration of IBPL was followed by faster absorption and elimination of R and S enantiomers. However, significant differences at alpha=0.05 were observed only at the stage of elimination. AUC was markedly higher following administration of suppositories containing IBP than following suppositories with IBPL and this pertained to both R and S enantiomers. Evident inversion of R into S form was noted 30 min following IBPL administration and 1 h after IBP administration. Ionisation extent only insignificantly affected the scope of chiral inversion of ibuprofen R into S form (AUC(S-IBP)/AUC(R IBP)=1.66, AUC(S-IBPL)/AUC(R-IBPL)=1.57). No presystemic inversion of R into S was observed in rabbits following administration of IBP or IBPL in suppositories. IBP enantiomers were isolated from 0.5 ml serum using solid phase extraction in C(18) columns and were quantified by HPLC using the chiral Whelk O1 column and UV detector (lambda=264 nm). PMID- 10802410 TI - Characterisation of a novel solid lipid nanoparticle carrier system based on binary mixtures of liquid and solid lipids. AB - A drug carrier of colloidal lipid particles with improved payloads and enhanced storage stability was investigated. Based on the experiences with hard fats nanoparticles, a new type of solid lipid nanoparticles (SLN) has been developed by incorporating triglyceride containing oils in the solid core of said particle. The structure and mixing behaviour of these particles were characterised and practical implications on controlled release properties tested. Nanoparticles were characterised by their melting and recrystallisation behaviour as recorded by differential scanning calorimetry (DSC). Polymorphic form and bilayer arrangement were assigned by wide-angle X-ray scattering (WAXS) and small-angle X ray scattering (SAXS). Size distribution and storage stability were investigated by laser diffractometry (LD). Release properties were studied by drug release model according to Franz. A medium chain triglyceride oil was incorporated successfully in a matrix of a solid long chain glyceride. The crystal order was greatly disturbed, however, the carrier remained solid. The oil inside the particle remained in a liquid state and induced a slight shift form the beta' polymorph to the beta(i) form. Long spacings varied within 0.1 nm with increasing oil loads. Nanoparticles with low oil concentrations showed sustained release properties. Improved drug load levels were encapsulated by lipid particles supplemented with oily constituents. Thus, the presented carrier adds additional benefits to the well-known opportunities of conventional SLN and is suited for topical use. PMID- 10802411 TI - A new method of characterising liquid uptake within particles over short time periods. AB - A method of measuring both the capacity and rate of absorption of liquid by powders of small particles over short time periods (of the order of a few seconds or less) is presented. The method is based on the measurement of the isothermal transient ionic current in a sample cell containing the absorbant material and the liquid. The method has been tested on solid glass beads, porous glass beads and cellulose agglomerates. Properties such as the instantaneous absorption capacity and rate can be characterised within a few seconds. No other technique is currently able to measure these fast outcomes. PMID- 10802412 TI - Extrusion-spheronization of pH-sensitive polymeric matrix pellets for possible colonic drug delivery. AB - The aim of this study was to investigate extrusion-spheronization pelletization for preparing pH-sensitive matrix pellets for colon-specific drug delivery. The effects of three independent variables (amounts of Eudragit S, citric acid and spheronizing time) on pellet size, shape (roundness and aspect ratio), and drug release were studied with central composite design. The pellets contained ibuprofen as a model drug, citric acid as a pH-adjusting agent, Eudragit S as a pH-sensitive binder and microcrystalline cellulose (MCC). The pellets were prepared with Nica extrusion-spheronizing equipment and subsequently enteric coated using an air-suspension technique. Eudragit S as a pH-sensitive matrix former in pellets increased the pellet size and influenced pellet roundness. In small amounts Eudragit S increased pellet roundness but in larger amounts pellet roundness was reduced. Citric acid promoted the pelletization process resulting in a narrower area distribution. The pH-sensitive matrix pellet failed to delay the drug release. The combination of citric acid and enteric coating, however, delayed the drug release for 15 min in a pH 7.4 phosphate buffer. PMID- 10802413 TI - Matrix mini-tablets based on starch/microcrystalline wax mixtures. AB - Matrix mini-tablets based on a combination of microcrystalline waxes and starch derivatives were prepared using ibuprofen as a model drug. The production of mini tablets was preferred over the production of pellets, as up-scaling of the pelletisation process seemed problematic. Prior to tabletting, melt granulation in a hot stage screw extruder and milling were required. The in vitro drug release was varied using microcrystalline waxes with a different melting range, the slowest drug release being obtained with a formulation containing a microcrystalline wax with a melting range between 68 and 72 degrees C. Generally speaking increasing the wax concentration resulted in a slower drug release. In vitro drug release profiles were also modified using different starches and mixtures of starches. Increasing the ibuprofen concentration to 70% resulted in a faster drug release rate. PMID- 10802414 TI - Dynamic laser speckle pattern in monitoring of local deformation of tablet surface after compression. AB - A method and a sensor device were developed to detect dynamic laser speckle pattern for monitoring of deformation of pharmaceutical tablets after compression. Temporal variations in optical signal revealed local surface deformation of the ibuprofen and the starch acetate tablets as well as anisotropy of tablet surfaces at the inspection location. PMID- 10802416 TI - Telephone contact of patients visiting a large, municipal emergency department: can we rely on numbers given during routine registration? AB - We sought to determine whether we could successfully contact patients for follow up using telephone numbers given during routine emergency department (ED) registration. Every fifth patient visiting our ED during the study period was eligible. Three calls were made to each number. Calls began 7 days after the ED visit. Of 1,136 patients, we successfully contacted 478 (42.1%). Of those patients unreachable across all three attempts, 183 (16.1%) had given wrong numbers, 133 (11.7%) had disconnected lines, and 156 (13.7%) had three consecutive "no answers." Females and patients with nonurgent complaints were significantly more likely to be contacted. Despite stringent calling protocols, we successfully contacted only 42% of our patients. Nearly 28% gave wrong or disconnected numbers. Placing two additional calls to those patients who were not home or did not answer initially nearly doubled the overall contact rate, although similar efforts for patients who initially gave wrong or disconnected numbers yielded no appreciable gains. Females and nonurgent patients were over represented. PMID- 10802415 TI - Vasopressin, renin, and adrenocorticotropic hormone levels during the resuscitation of hemorrhagic shock in dogs. AB - Hemorrhage is a potent stimulus for the release of vasopressin (VP), renin, and adrenocorticotropic hormone (ACTH). The goal of this study was to analyze changes in plasma VP, renin, and ACTH levels during hemorrhagic shock and resuscitation with two different solutions: hypertonic acetate dextran (HAD) and lactated Ringer's (LR) solution. Eight randomized dogs were shocked by removing 37 +/- 9 mL/kg of blood while maintaining the mean arterial pressure (MAP) at 45 +/- 5 mmHg for 1 h. Test solutions were randomized and infused as needed with the hemorrhaged blood to restore the MAP and cardiac index to baseline. Blood samples for hormone analyses were taken in baseline, shock, and resuscitation periods. For each experiment, all hormone levels increased in the postshock period and then returned to baseline values after resuscitation with both solutions. VP and renin levels rapidly returned to baseline values after resuscitation in the LR dogs compared with the HAD dogs (p < 0.05). By contrast, there was no significant difference in ACTH levels between the two solutions. High-volume infusion with LR achieves more rapid restoration than small-volume infusion with HAD for VP and renin levels. PMID- 10802417 TI - Munchausen syndrome presenting acutely in the emergency department. AB - Munchausen syndrome is a rare psychiatric disorder in which patients purposely harm themselves to gain medical attention. These patients may present to unsuspecting emergency department (ED) staff members with life-threatening signs and symptoms. We report a case of Munchausen syndrome in which a young man with ties to the medical community and a history of substance abuse presented to our ED with refractory hypoglycemia and a dilated right pupil. The patient required large amounts of dextrose, endotracheal intubation, mechanical ventilation, and admission to the intensive care unit. Further investigation revealed that he had purposely injected himself with a large dose of insulin and instilled atropine drops in his right eye for the purpose of seeking hospitalization. The history, details, diagnosis, treatment, and prognosis are discussed in the context of this case. PMID- 10802418 TI - Spontaneous splenic rupture due to subcutaneous heparin therapy. AB - We report a case of spontaneous splenic rupture in a 59-year-old woman who was receiving 15,000 units of heparin subcutaneously (s.c. ) twice a day for deep venous thrombosis (DVT) prophylaxis. Her past medical history included multiple DVT, pulmonary emboli, and ovarian cancer stage III-C with known ascites. The diagnosis of splenic rupture was initially missed because of the ascites. This case illustrates both a previously undescribed complication of s.c. heparin therapy and a failure of ultrasound diagnosis. We emphasize the unique presentation, difficulty in diagnosis, and need for early surgical involvement to ensure the most favorable outcome. PMID- 10802419 TI - Miller Fisher syndrome: an uncommon acute neuropathy. AB - The syndrome of ophthalmoplegia, ataxia, and areflexia was first described in 1956 by Miller Fisher. This syndrome has long been believed to be a variant of Guillain-Barre syndrome (GBS), mainly because of its areflexia, cerebrospinal fluid findings, and its postinfectious presentation. The case of an 11-year-old male with Miller Fisher syndrome (MFS) is described. MFS differs from GBS in several key clinical features and presents an extensive and challenging differential diagnosis. It is useful to recognize MFS as both a variant of GBS and a distinct entity with its own therapeutic considerations. PMID- 10802420 TI - Abdominal pain secondary to stump appendicitis in a child. AB - We report a case of an 11-year-old male who presented with abdominal pain and vomiting. The patient had a notable past medical history of having had an appendectomy at our institution 1 year previously. Because of progressive clinical signs of peritonitis, an exploratory laparotomy was performed and the patient was found to have stump appendicitis. The entity of stump appendicitis is always possible when evaluating patients with abdominal pain who have a history of appendectomy. PMID- 10802421 TI - Colchicine-induced bone marrow suppression: treatment with granulocyte colony stimulating factor. AB - Bone marrow aplasia is a frequent complication of colchicine poisoning. This typically occurs on day 3 to 5 postexposure, and the blood cell counts remain depressed for a week or more. Unfortunately, because patients suffering from colchicine toxicity develop multiple organ complications and sepsis, the morbidity and mortality associated with bone marrow depression is high. In this article, we present three cases of colchicine toxicity in which granulocyte colony-stimulating factor (G-CSF) was used to treat bone marrow depression. In all three cases, there was a dramatic increase in the white cell count and, to a lesser extent, the platelet count. In view of the critical nature of the bone marrow depression and multi-organ toxicity induced by colchicine, we believe that consideration of the use of G-CSF to shorten the duration of neutropenia is warranted. PMID- 10802422 TI - Acute cyanide poisoning: a case report. AB - A case of attempted homicide by cyanide ingestion is reported. The victim, a 19 year-old woman, unknowingly ingested cyanide and presented to the Emergency Department unresponsive, in shock, and in profound metabolic acidosis. The differential diagnosis of this presentation and the patient's successful treatment are reviewed. The important management issues surrounding the treatment of cyanide poisoning are discussed. PMID- 10802423 TI - Ketamine abusers presenting to the emergency department: a case series. AB - Ketamine hydrochloride, familiar to emergency physicians as a dissociative anesthetic, has been abused as a hallucinogen for almost 30 years. The drug produces effects similar to those of phencyclidine but with a much shorter duration of effect. Since 1996, an increasing number of patients have presented to Connecticut Emergency Departments (EDs) after the intentional abuse of ketamine. Because the medical literature contains almost no information on the consequences of ketamine abuse, we have compiled a series of ketamine abusers presenting to the ED. Among the 20 patients in this series, common presenting complaints included anxiety, chest pain, and palpitations. Tachycardia was the most common physical examination finding. Nystagmus, a common finding after phencyclidine use, was seen in only three cases. The most frequent complications after ketamine abuse were severe agitation and rhabdomyolysis. The symptoms of ketamine intoxication appear to be short-lived, with 18 of the 20 patients discharged from the ED within 5 h of presentation. Emergency physicians should include ketamine in the differential diagnosis of drug- or toxin-induced hallucinations. Methods for detecting this drug in biologic fluids are reviewed as are treatment recommendations for managing the patient who presents to the ED after abusing ketamine. PMID- 10802424 TI - Utilization of a glucagon infusion in the management of a massive nifedipine overdose. AB - This case report describes a continuous i.v. infusion of glucagon used to reverse the cardiovascular manifestations of a nifedipine overdose in a patient who presented after a massive nifedipine extended-release tablet ingestion. In this patient, glucagon appeared to be effective in the management of this toxicologic emergency. PMID- 10802425 TI - Cough and shortness of breath. PMID- 10802426 TI - Tricyclic antidepressant overdose and electrocardiographic changes. PMID- 10802427 TI - Amoebic abscess. PMID- 10802428 TI - Video analysis of emergency medicine residents performing rapid-sequence intubations. AB - The purpose of this study was to evaluate Emergency Medicine resident physicians' compliance with our institution's rapid sequence intubation (RSI) protocol by the use of videotape analysis. We conducted a prospective, observational study of Emergency Medicine resident physicians (EM 1,2,3) as they were videotaped performing RSI on medical and trauma patients. The videotapes were reviewed by the study investigators to assess the rates of deviation from our standard RSI protocol. Forty-four RSIs performed by 33 residents were studied. The most common deviations from our standard RSI protocol concerned proper use of the Sellick maneuver (45%) and use of the end-tidal CO(2) detector (34%). Videotape analysis provides an objective measure of Emergency Medicine resident performance of RSI. PMID- 10802429 TI - A survey of Canadians enrolled in American Emergency Medicine residencies. AB - Despite the existence of Emergency Medicine (EM) residency programs in Canada, Canadian physicians continue to pursue EM training in the United States. To determine the factors that may influence these Canadian physicians to return to practice in Canada, a survey was sent to all Canadians enrolled in U.S. EM training programs. Seventeen of 22 (77%) post-graduate trainees responded. Residents said they had chosen U.S. training mainly because of the low number of residents in Canadian EM specialty programs, and they also had the perception that U.S. EM training was superior. Lower salaries, restrictions on location of practice, and an inability to obtain Royal College certification were the factors most likely to prevent a return to Canada. Six of the 17 respondents (35%) said they were definitely or probably returning to Canada. Given the limited number of Canadian training positions and the Canadian Emergency Physician workforce shortfall, the U.S. training route appears to be underutilized. PMID- 10802430 TI - Do the hamstrings and adductors contribute to excessive internal rotation of the hip in persons with cerebral palsy? AB - Children with cerebral palsy frequently walk with excessive internal rotation of the hip. Spastic medial hamstrings or adductors are presumed to contribute to the excessive internal rotation in some patients; however, the capacity of these muscles to produce internal rotation during walking in individuals with cerebral palsy has not been adequately investigated. The purpose of this study was to determine the hip rotation moment arms of the medial hamstrings and adductors in persons who walk with a crouched, internally-rotated gait. Highly accurate computer models of three subjects with cerebral palsy were created from magnetic resonance images. These subject-specific models were used in conjunction with joint kinematics obtained from gait analysis to calculate the rotational moment arms of the muscles at body positions corresponding to each subject's internally rotated gait. Analysis of the models revealed that the medial hamstrings, adductor brevis, and gracilis had negligible or external rotation moment arms throughout the gait cycle in all three subjects. The adductor longus had an internal rotation moment arm in two of the subjects, but the moment arm was small (<4 mm) in each case. These findings indicate that neither the medial hamstrings nor the adductor brevis, adductor longus, or gracilis are likely to be important contributors to excessive internal rotation of the hip. This suggests that these muscles should not be lengthened to treat excessive internal rotation of the hip and that other factors are more likely to cause internally-rotated gait in these patients. PMID- 10802431 TI - Quantitative assessment of gait deviation: contribution to the objective measurement of disability. AB - Three biomechanical parameters based on force plate measurements were defined as indicators of gait deviation. Symmetry was specified as the relative difference in stance time and vertical impulse loading between both feet, constancy as the mean S.D. of the force curves for one subject under a specified gait condition and discrepancy as the average difference between the individual gait pattern and the expected force curves, normalised by the value of the S.D. in a control group. One hundred and forty four patients with osteoarthritis (OA) of the lower extremity and 144 control subjects were studied. There were 45 patients with OA of the hip, 54 of the knee and 45 of the ankle and their function was determined using the Harris Hip score, the hospital for special surgery knee score and the Mazur ankle score, respectively. The temporal asymmetry indicator was more sensitive to unilateral joint affliction, whereas the discrepancy indicators were sensitive to the presence of OA. Both correlated with the patient's function as measured by the relevant clinical score. A significant increase of gait discrepancy was detected in the arthritis group when patients were asked to walk at faster speeds; whilst walking barefoot led to an unexpected reduction of intra subject kinetic variability. Our results confirm the validity and usefulness of the gait deviation concept in patients with OA. PMID- 10802432 TI - Stride smoothness evaluation of runners and other athletes. AB - The purpose of this study was to compare an objective measurement of smoothness between a group of runners and a group of non-runners during running and fast walking. Smoothness was quantified by evaluating the endpoint jerk-cost (JC) at the heel. Subjects walked at a speed of 1.75 m.s(-1) and ran at a speed of 3.35 m.s(-1) on a motor driven treadmill while 2-D kinematic data (60 Hz) were collected from a sagittal plane view. The runners were found to be smoother than the non-runners during both gait conditions, suggesting that this group was inherently smoother in gait related tasks. This study demonstrated that the smoothness of gait can be quantified objectively by evaluating the end-point JC at the heel, and that competitive runners tend to exhibit smoother strides than recreational runners during both running and fast walking. PMID- 10802433 TI - Effect of plantar flexor muscle stiffness on selected gait characteristics. AB - Although plantar flexor muscle strength is known to affect gait characteristics, the effect of plantar flexor (PF) muscle stiffness on gait is unknown. The purpose of this study was to determine the effect of PF muscle stiffness, in addition to PF strength, on the gait characteristics of subjects with diabetes and peripheral neuropathy (DM & PN) and age-matched controls. Thirty-four subjects were tested (17 DM & PN, 17 controls), 10 males and seven females in each group. PF muscle strength (concentric peak torque) explained a significant amount of variance in all gait variables (20-37%). Passive stiffness added a unique contribution to plantar flexor peak moment and walking speed (10-11%). Plantar flexor stiffness, in addition to strength, may be an important factor contributing to gait characteristics. Treatment techniques designed to preserve or increase stiffness may be indicated for patients with limited PF strength. PMID- 10802434 TI - Effect of simulating leg length inequality on pelvic torsion and trunk mobility. AB - Although techniques such as roentgenograms and magnetic resonance imaging can provide definitive information about leg length inequality, they are not easily implemented for screening purposes. Using relative heights of palpated iliac crests as criteria for determining degree of lateral pelvic tilt, we examined the immediate effect of simulating leg length inequality on pelvic torsion and trunk flexion. In seven healthy men and 22 healthy women, 18-28 years of age, a lift of at least 15 mm was placed under either foot to laterally tilt the pelvis 1.2 degrees or more. In eight subjects with pre-existing lateral pelvic tilts of 1.8 degrees or more, a lift was also used to eliminate the tilt. We examined how this tilting affected torsion between the innominates and mobility of the trunk. The innominate contralateral to the lift became more anteriorly rotated than the ipsilateral innominate and lateral flexion of the trunk increased toward the side of the lift. Both of these effects can be associated with clinical leg length inequality, so a lateral pelvic tilt on the order of 1.2 degrees, if encountered in the clinic, should signal the suitability of more extensive examination for possible lower limb asymmetry. PMID- 10802435 TI - Common abnormal kinetic patterns of the knee in gait in spastic diplegia of cerebral palsy. AB - We studied the kinetic characteristics of the knee in patients with spastic diplegia. Twenty three children with spastic diplegia were recruited and had their 46 limbs categorised into the following four groups: jump (n=7), crouch (n=8), recurvatum (n=14) and mild (n=17). In the crouch pattern, the patients usually had a larger and longer lasting internal knee extensor moments in stance suggesting that rectus femoris had a relatively high activation. In the recurvatum pattern, the internal knee flexor moment was large and long lasting in stance. The biceps femoris showed less activity on EMG although the knee flexor moment was large and we concluded that the soft tissue behind the knee joint provided this flexor moment. In the jump knee pattern there was abnormal power generation at the knee and ankle joints in initial stance, which did not contribute to normal progression but aided upward body motion. In the mild group the kinetic data was similar to that seen in normal children. Knowledge of kinetic patterns in these patients may help in their subsequent management. PMID- 10802436 TI - The influence of external knee moments on the outcome of total meniscectomy. A comparison of radiological and 3-D gait analysis measurements. AB - Twenty one individuals who had undergone unilateral total meniscectomy, and had no significant antero-posterior (AP) laxity underwent three dimensional (3-D) gait analysis, bilateral radiological assessment of varus/valgus alignment and tibio-femoral osteoarthritis (TFOA). Average foot progression angle during the stance phase of gait was not related to knee adduction moment. The findings support the previously noted correlation between varus/valgus alignment and TFOA. The dynamic gait parameter of the adduction moment in early stance did not correlate with either hip-knee-ankle angle or TFOA, thus the expected influence of external moments on outcome after meniscectomy was not found. Thus the correlation between varus/valgus alignment and TFOA can not be explained by a simple loading algorithm. PMID- 10802437 TI - Preparatory adjustments during gait initiation in 4-6-year-old children. AB - The preparatory adjustments related to gait initiation in a group of six children (4-6 years old) were studied in comparison to a group of six adults (50-61 years old). Muscle activity, ground reaction forces and body kinematics were recorded during the initiation of gait in subjects standing with one foot on each of two forceplates. Like adults, children had consistent anticipatory activations of the tibialis anterior (TA) muscle accompanied by center of pressure (COP) displacements, but the relative magnitude of both the backward COP displacement and concomitant initial TA activation burst were lower (P<0.03 and P<0.001 respectively) in children. The latter findings were associated with a lower rate of forward progression in children (P=0.01), and a lack of significant covariance between the backward shift and forthcoming velocity. In contrast, the preparatory adjustments in the medio-lateral (M/L) direction were prominent in children. Larger M/L peak force rates in children (P=0. 01) were associated with an earlier (P=0.007) weight transfer to the stance limb; moreover, children initiated gait from a wider (P=0.04) base of support and had a trajectory of the total COP that was lateral rather than posterior like in adults. The consistent preparatory adjustment responses indicate that the motor program for initiating gait is functional at this age. The prominence of the preparatory adjustments in the M/L direction together with a reduced magnitude of the responses in the antero posterior direction suggest that the anticipatory behavior for initiating gait develops first in the frontal plane and that more walking experience and better postural stability are required to fully achieve the adult-like control of the gait initiation process. PMID- 10802438 TI - Changes in gait patterns in 10-year-old boys with increasing loads when walking on a treadmill. AB - This study examined the gait pattern, heart rate and blood pressure in children carrying school bags of 0 (as control), 10, 15 and 20% of their own body weight whilst walking on a treadmill. When compared to the 0% load condition, the 20% load condition induced a significant increase in trunk forward lean, double support and stance duration, and decreased trunk angular motion and swing duration and a prolonged blood pressure recovery time. The 15% load condition induced a significant increase in trunk forward lean and prolonged blood pressure recovery time. No significant difference was found in the measured parameters between the 10 and 0% load conditions. We concluded that back pack weight should not exceed 10% of body weight in 10-year-old boys. PMID- 10802445 TI - Temporal and spatial determination of EEG-seizure onset in the frequency domain. AB - OBJECTIVE: A quantitative analysis of scalp electric fields in patients suffering from pharmacoresistant temporal lobe epilepsy was performed in order to study the development of rhythmic ictal activities over time. METHODS: A method that calculates phase-corrected voltage maps in the frequency domain (FFT approximation) was applied to ictal multichannel recordings in 10 epileptic patients. The onset of the ictally dominant frequency was determined and its temporal evolution over a time period of 46 s around the ictal EEG onset was studied. The analysis was completed by a linear inverse solution that estimated the sources of the dominant frequency. RESULTS: This method permitted the identification of an ictally dominant frequency which started on the average prior to the onset of initial EEG signs as determined by visual inspection. The frequency incremented during the evolution of the seizure in all patients. The linear inverse solution algorithm localized the source of this frequency to the brain region which was clinically determined as the site of seizure onset and whose resection rendered all patients seizure-free. CONCLUSIONS: Our data suggest that the constant increase of the ictally dominant frequency is related to the amount of temporal lobe tissue generating the ictal discharges. Frequential analysis of ictal electric fields can be reliably used to detect focal pathological activity early during seizure onset arising in deep structures such as the mesial temporal lobe. PMID- 10802446 TI - Using weighted linear spatial decomposition to investigate brain activity through a set of fixed current dipoles. AB - OBJECTIVES: We developed a method with the aim of decorrelating scalp EEG based on a set of spatial constraints. METHODS: We assume that the scalp EEG can be modelled by a small number of current dipoles of fixed location and orientation, placed at regions of interest. The algorithm is based on weighted linear spatial decomposition in order to obtain a weighted solution to the inverse problem. An EEG data matrix is first weighted in favour of a single dipole in the set. The dipole moment is then calculated from the weighted EEG by the pseudo-inverse method. This is repeated for each dipole. RESULTS: Six seizures were processed from 4 patients using the standard least-squares solution and our weighted version. The average cross-correlation between channels was calculated for each case. The first method resulted in a mean drop in cross-correlation of 16.5% from that of the scalp. Our method resulted in a reduction of 34.5%. CONCLUSIONS: Our method gives a more spatially decorrelated signal in regions of interest (although it is not intended as an accurate localization tool). Subsequent analysis is more robust and less likely to be dependent on specific recording montages. This is more than could be obtained using a standard least-squares solution using the same model. PMID- 10802447 TI - Theta oscillations and the ERP old/new effect: independent phenomena? AB - OBJECTIVES: The hypothesis is examined whether a memory-related change in induced band power (oscillatory old/new effect) is functionally related to a memory related increase in ERP positivity (ERP old/new effect). METHODS: In order to avoid a confounding on the measurement level, induced band power (IBP) was used as a measure that is devoid of the influence of evoked components. The EEG was recorded during a recognition memory task. RESULTS: The results show that compared to correctly rejected words, targets (remembered words) elicit a significantly larger P300. An oscillatory old/new effect was found for the delta and theta but not for the alpha band. It is manifested by an increase in delta and theta IBP which is significantly larger for targets than for correctly rejected words. It can be observed during the same time interval and shows the same topographic distribution as the ERP old/new effect. Most importantly, however, the ERP old/new effect (as well as the P300 itself) is generated by very slow frequencies which lie below the delta band. CONCLUSIONS: These findings demonstrate that the two types of old/new effects are functionally related. Possible physiological mechanisms underlying this relationship are discussed in terms of a threshold change in the cortex (generating the P300) that occurs during an increase in hippocampal theta activity (generating an increase in induced theta power). PMID- 10802448 TI - Direct demonstration of the effect of lorazepam on the excitability of the human motor cortex. AB - OBJECTIVES: The present study explored the effects of lorazepam, a benzodiazepine with agonist action at the GABA(A) receptor, on human motor cortex excitability as tested using transcranial magnetic stimulation. METHODS: We recorded directly the descending volley evoked by single and paired transcranial magnetic stimulation from the spinal cord of a conscious subject with a cervical epidural electrode before and after a single oral dose of lorazepam. We evaluated the effects of lorazepam on the descending volleys evoked by a single magnetic stimulation and paired cortical stimulation using the intracortical inhibition paradigm (subthreshold conditioning stimulus) and the short latency intracortical facilitation paradigm (suprathreshold conditioning stimulus). RESULTS: Using a single magnetic stimulus lorazepam decreased the amplitude of the later I waves in the descending volley; this was accompanied by a decrease in the amplitude of the evoked EMG response. Using the intracortical inhibition paradigm lorazepam increased the amount of corticocortical inhibition, particularly at 4 and 5 ms interstimulus intervals. There was no effect on the amount of facilitation observed in the short latency intracortical facilitation paradigm. CONCLUSIONS: The present findings provide direct evidence that lorazepam increases the excitability of inhibitory circuits in the human motor cortex. PMID- 10802449 TI - Modulation of corticospinal excitability by repetitive transcranial magnetic stimulation. AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) is able to modulate the corticospinal excitability and the effects appear to last beyond the duration of the rTMS itself. Different studies, employing different rTMS parameters, report different modulation of corticospinal excitability ranging from inhibition to facilitation. Intraindividual variability of these effects and their reproducibility are unclear. METHODS: We examined the modulatory effects of rTMS to the motor cortex at various frequencies (1, 10, 20 Hz) and at different time-points in twenty healthy volunteers. RESULTS: We observed significant inhibition of MEPs following 1 Hz rTMS and significant facilitation of MEPs following 20 Hz rTMS for both day1 and day 2. Interestingly, at 1 Hz and 20 Hz rTMS, the modulatory effect produced by rTMS was greater on day 2. However, there was no significant change in corticospinal excitability following 10 Hz rTMS neither on day 1 nor day 2. CONCLUSION: Our findings raise questions as to how stimulation parameters should be determined when conducting studies applying rTMS on multiple days, and in particular, studies exploring rTMS as a treatment modality in neuropsychiatric disorders. PMID- 10802450 TI - Tremor-correlated cortical activity detected by electroencephalography. AB - OBJECTIVE: In this study we investigated whether cortical activity related to Parkinsonian resting tremor can be detected by electroencephalography (EEG). METHODS: Seven patients with idiopathic Parkinson's disease suffering from unilateral tremor participated in the study. Electromyography (EMG) signals arising from the wrist extensor and flexor muscles as well as a high resolution EEG were recorded simultaneously. Coherencies between EEG and EMG were calculated. RESULTS: In all patients, we found highly significant coherencies at the tremor frequency or its first harmonic between the tremor EMG and contralateral EEG channels. There were no significant coherencies between the tremor EMG and ipsilateral EEG channels. Isocoherency maps illustrating the topography of the coherencies over the scalp showed that the maximum coherencies were situated over the cortical motor areas. In one case, a high coherency was also found over the parietal cortex. CONCLUSIONS: The results show for the first time that tremor-correlated cortical activity can be detected by electroencephalography. The findings underline that motor areas of the cerebral cortex are involved in the neuronal network generating resting tremor in Parkinson's disease. PMID- 10802451 TI - Wavelet decomposition of the blink reflex R2 component enables improved discrimination of multiple sclerosis. AB - OBJECTIVES: The blink reflex R2 component was subjected to wavelet decomposition for time feature extraction in order to classify the functional status of patients with multiple sclerosis. METHODS: The blink reflex was recorded bilaterally with unilateral stimulation of the supra-orbital nerve in 37 normal subjects and 9 patients with multiple sclerosis (MS). The late component, R2, was subjected to time-frequency decomposition using the Daubechies-4 wavelet. Using the time-frequency coefficients, the mean time of the R2 wave as well as the standard deviation of the R2 interval were calculated in each trial. The wavelet transform enables noise reduction by allowing selective use of frequency bands with high signal-to-noise ratio for time feature extraction; therefore automatic estimation of time parameters is robust. The distribution densities of the mean and the standard deviation of the R2 wave duration for the set of trials for each subject were computed. RESULTS: An appreciable difference in the densities of the two parameters extracted in the wavelet domain was seen between normals and patients. This is in contrast to the onset latency of R2 which poorly discriminates MS patients from normals. CONCLUSION: The results suggest that the mean and standard deviation of the R2-time robustly estimated using wavelet decomposition can be used to support clinical diagnosis in tracking the functional status of patients with diseases like multiple sclerosis. PMID- 10802452 TI - Phrenic nerve conduction study in demyelinating neuropathies and open-heart surgery. AB - OBJECTIVES: The aim of this study was to determine normal values of phrenic nerve conduction (PNC) in healthy individuals; to evaluate the subclinical extent of phrenic nerve involvement in Guillain-Barre syndrome (G-B) and hereditary motor and sensory neuropathy-I (HMSN-I), and to evaluate phrenic nerve damage after cardiac surgery. MATERIALS AND METHODS: PNC was performed by transcutaneous stimulation in the neck and recording the diaphragmatic potential from surface electrodes placed at the seventh and eight intercostal spaces. PNC was performed bilaterally in 25 healthy volunteers and 25 patients before and after open-heart surgery. Right PNC was also performed in 5 cases with G-B and 5 patients with HMNS-I. RESULTS: Latency and amplitude of the diaphragmatic potential were the same in controls and in patients with cardiac disease before surgery. After surgery, 28% of patients had left phrenic nerve inexcitability, and 8% had reduced amplitude of the response. These 9 patients demonstrated elevation of the left hemidiaphragm on chest radiography. Left PNC performed 1 year after the operation showed improvement in latency and amplitude of the responses in all except one patient. PNC was prolonged in 4 out of 5 cases with G-B and in all patients with HMNS-I. CONCLUSIONS: PNC is an easy and reliable method in evaluating phrenic nerve damage due to hypothermia or primary stretch injury in patients after cardiac surgery. PNC may be helpful in detecting diaphragmatic involvement before clinical ventilatory insufficiency in demyelinating neuropathies such as G-B and HMNS-I. PMID- 10802453 TI - Automatic carpal tunnel syndrome tester. AB - OBJECTIVE: A device for automatic testing of sensory latency across the carpal tunnel is described. A fixed bar contains a stimulating electrode distally and a bipolar recording set up with the two poles at 7 and 14 cm distance proximally. The device is thus based on the so called 14/7 method, using orthodromic testing. METHODS: Automatic testing of electrode resistance and muscle relaxation precedes the self triggered start of stimulation. The latencies are analyzed according to a special algorithm. The proximal/distal latency index is increased in carpal tunnel syndrome (CTS). RESULTS: Healthy controls (32 hands in 32 subjects) and patients referred with suspicion of CTS (258 hands) were studied with the new device, by conventional technique and sometimes by inching (centimetering) across the carpal tunnel. A positive CTS result with the tester was in accordance with the routine test in 94% of the patients. Conversely, routine studies showed CTS in 118 hands. Among those, the tester agreed in 44, showed no signals in 70 and indicated normal findings in 4 hands. In cases with a combination of polyneuropathy and CTS, the CTS was detected in 5 out of 6 cases. A complete test takes about 1 min per hand. CONCLUSIONS: The results are very promising. PMID- 10802454 TI - Schizophrenia-like deficits in auditory P1 and N1 refractoriness induced by the psychomimetic agent phencyclidine (PCP). AB - OBJECTIVES: The amplitude of the cortically generated auditory event-related potential (ERP) components P1 and N1 decreases as the interval between successive stimuli (ISI) decreases. Although the phenomenon of P1 and N1 refractoriness is well established, the underlying mechanisms are poorly understood. The present study investigates P1 and N1 refractoriness in the awake monkey in order to investigate underlying mechanisms. METHODS: Auditory ERP were obtained in response to repetitive auditory stimuli presented at 5 levels of ISI between 150 ms and 9 s, prior to and following administration of the selective N-methyl-D aspartate (NMDA) antagonist phencyclidine (PCP). RESULTS: P1 and N1 amplitude declined in monkeys with decreasing ISI, with similar temporal characteristics to that observed in humans. PCP inhibited P1 and N1 generation at long, but not short, ISI producing a pattern similar to that recently observed in schizophrenic subjects. CONCLUSIONS: The present findings suggest that the primate P1/N1 model may be useful for investigating mechanisms underlying impaired information processing in schizophrenia, and that NMDA receptor dysfunction may play a key role in information processing dysfunction associated with schizophrenia. PMID- 10802455 TI - Schizophrenia: reduced signal-to-noise ratio and impaired phase-locking during information processing. AB - OBJECTIVE: This study was performed in order to clarify the mechanisms which underlie the reduced signal-to-noise of event-related potentials in schizophrenic patients. Specifically, we wanted to find out, whether it is reduced activation and/or synchronization (phase-locking) in specific frequency bands of the ongoing EEG which is related to the decreased signal amplitude and signal-to-noise ratio in schizophrenics. METHODS: We investigated 41 unmedicated schizophrenics (10 of them drug-naive) and compared them with healthy control subjects (n = 233) as well as unmedicated subjects with schizotypal personality (n = 21), who were considered to be high-risk subjects for schizophrenia, and unmedicated depressive patients (n = 71). We measured event-related activity during an acoustical choice reaction paradigm and calculated the signal-to-noise ratio, signal power and noise for a time interval of 50-200 ms after stimulus presentation. Signal-to noise ratio was calculated from the power of the averaged trials (signal power) divided by the mean power of the single trials minus the power of the average (noise power). Also, we performed a frequency analysis of the pre- and poststimulus EEG based on a factor analytical approach. Group comparisons were performed with ANCOVA. RESULTS: As expected, a decreased signal-to-noise ratio of evoked activity was found in the schizophrenic and a non-significant trend in the schizotypal subjects and the depressive patients. We were able to show that the observed decrease is due to a reduced signal power and an increase of absolute noise power. Frequency analysis of the evoked activity revealed that normals, schizophrenics schizotypal subjects and depressive patients increased theta/delta activity between pre- and poststimulus interval to a similar extend. However, this theta/delta-augmentation does not correlate with signal power in schizophrenics. Also, normals and depressive subjects augment coherence between both temporal lobes during information processing, which is not found in schizophrenics and schizotypal subjects. In contrast, these two groups augment frontal lobe coherence, which goes along with an increase of noise. CONCLUSIONS: Reduced stimulus-induced phase-locking and bitemporal coherence of cortically evoked activity but not a failure to activate the cortex may be responsible for the observed low signal-to-noise ratio during information processing in schizophrenics. Accordingly, schizophrenics increase noise after stimulus presentation instead of building up a signal. This is discussed in the framework of the theory of stochastic resonance. PMID- 10802456 TI - Steady-state visually evoked potential topography during the continuous performance task in normal controls and schizophrenia. AB - OBJECTIVES: To examine the latency topography of the steady-state visually evoked potential (SSVEP) in patients diagnosed with schizophrenia and normal controls while undertaking a visual vigilance task. METHODS: Twenty patients diagnosed with schizophrenia and 18 normal controls performed the A-X version of the continuous performance task (CPT A-X) where subjects are required to press a micro-switch on the unpredictable appearance of an 'X' that had been preceded by an 'A.' Brain electrical activity was recorded from 64 scalp sites and a 13 Hz spatially uniform visual flicker presented with the task was used to elicit a steady-state visually evoked potential (SSVEP). RESULTS: Following the appearance of the 'A' and 'X,' the control group demonstrated a transient SSVEP latency reduction at parietal and prefrontal sites. By contrast, the patients group showed no such SSVEP latency reduction. The prefrontal SSVEP latency changes in the 500 ms interval following the appearance of the 'X' were correlated with mean individual reaction time in both populations. CONCLUSIONS: We suggest that the SSVEP latency reduction may index excitatory processes and that the absence of prefrontal SSVEP latency reduction in schizophrenic patients may be a manifestation of reduced prefrontal activity or 'hypofrontality' observed with other neuroimaging modalities. PMID- 10802457 TI - Event-related desynchronization during an auditory oddball task. AB - OBJECTIVE: The present study addressed what kind of mental processes would be presented by the event-related desynchronization (ERD) relevant to the stimuli of an auditory oddball count task. METHODS: Electroencephalogram (EEG) was recorded from nine healthy subjects while target tones (2000 Hz, P = 0.2) and non-target tones (1000 Hz, P = 0.8) were presented randomly with constant stimulus onset asynchrony (SOA) of 3.3 s. To improve time resolution of ERD analysis, obtained EEG epochs were digitally convoluted by Gabor wavelet and averaged respectively. RESULTS: For target stimulus, prominent ERD was observed in left parieto occipital areas (peak latency: 400-600 ms), but there were no significant ERD for non-target stimulus. CONCLUSION: Our result suggests that magnitude of ERD would reflect amount of mental effort which was associated with intentional and voluntary processes rather than automatically sensory process. PMID- 10802458 TI - The mismatch negativity during natural sleep: intensity deviants. AB - Auditory event-related potentials (ERPs) were recorded from 7 subjects who slept for a single night in the laboratory. An 'oddball' sequence of brief tone pips, differing in intensity, was used. Frequently occurring 70 dB 'standards' were presented with infrequent 80 dB intensity increment deviants and 60 dB intensity decrement deviants. The probability of each deviant was 0.1. Stimuli were presented in a random sequence every 600 ms while subjects were awake but inattentive and during stages 2 and REM of sleep. During wakefulness, the intensity increments elicited a broad fronto-central negativity with two discernable peaks. The first, peaking at approximately 120 ms, showed a polarity inversion at the mastoid and likely represented a summation of the N1 wave and the mismatch negativity (MMN). The second, peaking at approximately 330 ms, may have reflected an enhanced N2b component. In REM sleep, the increment deviants elicited a small amplitude 100-200 ms negativity but its amplitude was not significantly larger than the baseline level. It was followed by a larger and significant 300-450 ms negativity but this was considered too delayed to represent the MMN. The decrement deviants elicited a small amplitude, but statistically non-significant, MMN-like wave during both wakefulness and in REM sleep. A MMN-like wave was absent in stage 2 sleep. PMID- 10802459 TI - Preprogramming and control activity of bimanual self-paced motor task in Parkinson's disease. AB - OBJECTIVE: The authors investigated programming (Bereitschaftspotential or BP) and control activity (Skilled Performance Positivity or SPP) of a bimanual, sequential, skilled motor act in off-therapy Parkinson's disease (PD) patients. METHODS: We recorded Movement Related Potentials (MRPs) in 12, non-demented, off therapy parkinsonian patients and in 17 control subjects who were performing a skilled, time-locked motor act, which was not routine in their everyday life but had to be learned: the Skilled Performance Task (SPT). BP, SPP and correct performances were evaluated in grand average waveforms and in sequential blocks. RESULTS: The analysis of correct performances showed that accuracy in PD patients was significantly lower than in the control group and this accuracy did not improve throughout the blocks. A significantly low level of performances was associated with an increased BP amplitude (P<0.05) and decreased SPP amplitude (P<0.05) in PD patients. CONCLUSION: Our findings suggest that skill motor learning is impaired in non-demented unmedicated PD patients. We discuss the view that PD patients may allocate more attentional resources, as suggested by the increased BP amplitude, the decreased SPP amplitude and the low correct performances, in order to perform a new skilled motor act. PMID- 10802460 TI - Oscillatory frontal theta responses are increased upon bisensory stimulation. AB - OBJECTIVES: To investigate the functional correlation of oscillatory EEG components with the interaction of sensory modalities following simultaneous audio-visual stimulation. METHODS: In an experimental study (15 subjects) we compared auditory evoked potentials (AEPs) and visual evoked potentials (VEPs) to bimodal evoked potentials (BEPs; simultaneous auditory and visual stimulation). BEPs were assumed to be brain responses to complex stimuli as a marker for intermodal associative functioning. RESULTS: Frequency domain analysis of these EPs showed marked theta-range components in response to bimodal stimulation. These theta components could not be explained by linear addition of the unimodal responses in the time domain. Considering topography the increased theta-response showed a remarkable frontality in proximity to multimodal association cortices. Referring to methodology we try to demonstrate that, even if various behavioral correlates of brain oscillations exist, common patterns can be extracted by means of a systems-theoretical approach. CONCLUSIONS: Serving as an example of functionally relevant brain oscillations, theta responses could be interpreted as an indicator of associative information processing. PMID- 10802461 TI - Characteristics of the human contra- versus ipsilateral SII cortex. AB - OBJECTIVES: In order to study the interaction between left- and right-sided stimuli on the activation of cortical somatosensory areas, we recorded somatosensory evoked magnetic fields (SEFs) from 8 healthy subjects with a 122 channel whole-scalp SQUID gradiometer. METHODS: Right and left median nerves were stimulated either alternately within the same run, with interstimulus intervals (ISIs) of 1.5 and 3 s, or separately in different runs with a 3 s ISI. In all conditions 4 cortical source areas were activated: the contralateral primary somatosensory cortex (SI), the contra- and ipsilateral secondary somatosensory cortices (SII) and the contralateral posterior parietal cortex (PPC). RESULTS: The earliest activity starting at 20 ms was generated solely in the SI cortex, whereas longer-latency activity was detected from all 4 source areas. The mean peak latencies for SII responses were 86-96 ms for contralateral and 94-97 ms for ipsilateral stimuli. However, the activation of right and left SII areas started at 61+/-3 and 62+/-3 ms to contralateral stimuli and at 66+/-2 and 63+/-2 ms to ipsilateral stimuli, suggesting a simultaneous commencing of activation of the SII areas. PPC sources were activated between 70 and 110 ms in different subjects. The 1.5 s ISI alternating stimuli elicited smaller SII responses than the 3 s ISI non-alternating stimuli, suggesting that a considerable part of the neural population in SII responds both to contra- and ipsilateral stimuli. The earliest SI responses did not differ between the two conditions. There were no significant differences in source locations of SII responses to ipsi- and contralateral stimuli in either hemisphere. Subaverages of the responses in sets of 30 responses revealed that amplitudes of the SII responses gradually attenuated during repetitive stimulation, whereas the amplitudes of the SI responses were not changed. CONCLUSIONS: The present results implicate that ipsi- and contralateral SII receive simultaneous input, and that a large part of SII neurons responds both to contra- and ipsilateral stimulation. The present data also highlight the different behavior of SI and SII cortices to repetitive stimuli. PMID- 10802463 TI - Alpha burst activity during human REM sleep: descriptive study and functional hypotheses. AB - OBJECTIVES: Spectral power contribution in the range of alpha activity is a well known electrophysiological feature of human REM sleep, which could be caused by the spontaneous bursts of alpha activity not associated with arousals that usually appear during this brain state. The present study was undertaken to determine the density of alpha bursts during tonic and phasic (oculomotor) REM periods for each REM cycle. In addition, this phasic brain event was also described from a spectral and topographical point of view. METHODS: Ten healthy right-handed subjects (5 females) aged 19-25 years (mean 22.9 years, SD 2.6) participated in the present study. Each selected subject filled in a daily sleep log for 2 weeks before the experimental night to provide information on all 3 salient aspects of sleep pattern, sleep experience and sleep effects. RESULTS: The results revealed that transient REM-alpha bursts, which lasted about 3 s and were accompanied by no increase in the EMG amplitude, appeared in all subjects who participated in this study, showing a higher density in the third and fourth REM cycle during phasic in comparison with tonic periods. The bandpass filtered signals showed the highest spectral contribution for the slower alpha components (8-9 Hz), the occipital scalp regions being the main generator source of this brain activity. CONCLUSIONS: The authors hypothesize that REM-alpha bursts may work as micro-arousals (or incomplete arousals) facilitating the brain connection with the external world in this cerebral state, whereas REM-alpha arousals - usually longer and accompanied by changes in the EMG amplitude - generate a shift of brain state associated with sleep fragmentation (complete arousal). PMID- 10802462 TI - Somatosensory evoked potentials in very preterm infants. AB - OBJECTIVE: Cross-sectional and longitudinal reference values of cortical N(1) peak latency of the median nerve SEP in very preterm infants. METHODS: In infants in a placebo control group within an L-thyroxine supplementation trial, born at less than 30 weeks' gestation, cortical N(1) peak latency was measured at 2 weeks, at term and at 6 months corrected age. Cross-sectional N(1) latency values obtained in 50 infants and complete series of longitudinal values obtained in 15 infants were analyzed in relation to postmenstrual age (PMA). RESULTS: Mean N(1) latency decreased from 66 ms at 2 weeks to 38 ms at term and 20 ms at 6 months corrected age. Possible confounding factors did not have any significant effect on N(1) latency at 2 weeks or at term age except cranial ultrasound abnormalities at 2 weeks of age. CONCLUSIONS: Longitudinal N(1) latency values were consistent with cross-sectional N(1) latency values. The observed N(1) latency at term and at 6 months corrected age suggest that extrauterine maturation of the somatosensory pathway in infants born at less than 30 weeks' gestation is delayed by extrauterine life. PMID- 10802464 TI - Sleep-EEG modulation of interictal epileptiform discharges in adult partial epilepsy: a spectral analysis study. AB - INTRODUCTION: In order to define accurately the relationship between EEG components (spindles, delta and theta frequencies) and the occurrence of interictal epileptiform discharges (IED) during sleep in partial epilepsy, a correlation study between spike overnight distribution and EEG spectral power time series was performed. METHODS: Eighteen patients (mean age: 24.7+/-5.5 years) affected by partial epilepsy underwent continuous EEG-polysomnography. The temporal series of Slow Wave Activity (SWA), Sigma Activity (SA) and Theta Band (TB), derived from spectral analysis, were obtained from a spike-free and pathologic alteration-free derivation, contralateral to the most active lead, where the IED count was performed. Relationships between SA, SWA and TB and time series of IED were tested by means of correlation techniques after data normalization. RESULTS: Our results revealed a significantly higher correlation between IED and SWA in 12 subjects; a significantly higher correlation between IED and SA in three subjects and a significant correlation with TB in three cases. CONCLUSIONS: Data suggest that in most adult patients with partial epilepsy IED production during sleep is facilitated by the action of synchronizing mechanisms which are active during NREM sleep and lead to the appearance of EEG delta waves. Nevertheless evidence is given of two smaller groups of patients. In one of them IED are more sensitive to the promoting action of the spindle generating mechanism, active during stage 2 of NREM sleep. In the other one the promoting action of TB, characterizing EEG during stage 1 and REM sleep, is evident. PMID- 10802465 TI - Asymmetric interhemispheric delta waves during all-night sleep in humans. AB - OBJECTIVE: In order to better understand the asymmetry of brain function during sleep, period-amplitude analysis of delta EEG activity was performed on polysomnograms (PSGs) in normal humans. Twenty healthy, right-handed male volunteers aged 22-35 years (mean age 27.2 years) served as subjects in this study. METHODS: EEGs were recorded from disc electrodes placed at bilateral frontal, central, parietal, occipital, anterotemporal and posterotemporal (10-20 electrode system) sites using A1+A2 for reference. Period-amplitude analysis was performed by the zero-crossing method using the Medilog Sleep Analyzing Computer. RESULTS: Delta counts in the right frontal and central regions during all-night sleep were significantly greater than in those of the left; total delta counts of the right frontal region were greater than those of the left in 18 of the 20 subjects. There were no significant differences in delta counts between the left and right hemispheres in parietal, occipital, anterotemporal, and postero temporal regions. CONCLUSIONS: These results suggest distinct laterality in the number of delta waves in the frontal and central regions, reflecting functional asymmetry of the brain during sleep. PMID- 10802466 TI - Sleep architecture, slow wave activity, and sleep spindles in adult patients with sleepwalking and sleep terrors. AB - OBJECTIVES: A very strong SWS intensity reflected by both an increased level of SWA and an abnormal sleep spindles distribution would be responsible for the major difficulty of parasomniac subjects in waking up from SWS, leading to episodes of parasomnia. METHODS: Eleven adult parasomniac subjects, 6 females and 5 males, with sleepwalking (SW) and/or sleep terrors (ST) and 11 age- and sex matched control subjects underwent polysomnography (PSG) during 2 consecutive nights. After an habituation and selection night followed by a 16 h period of controlled wakefulness, the sleep EEGs of the parasomniac and control subjects were analyzed on the second night by computer-aided visual scoring (integrated digital filtering analysis, IDFA) and spectral analysis (fast Fourier transform, FFT). Throughout the night subject behaviour was controlled and recorded by means of a video infra-red camera and videotape recorder. RESULTS: Fifteen episodes of parasomnia were recorded during the second night in the 11 subjects. Sleep analysis showed significantly (P<0.05) decreased sleep efficiency and stage 2 sleep (absolute values and percentage of total sleep time) and increased (P<0.05) slow wave sleep (absolute values and percentage of total sleep time). Arousal index and wake-time after sleep onset were significantly higher in parasomniac subjects. Sleep fragmentation was mainly concentrated in stages 3 and 4. The slow wave activity (SWA) absolute values averaged during the 2 min immediately preceding an episode of parasomnia were significantly higher than the SWA averaged during 2 min in the same stage 10 min before an episode of parasomnia. Moreover, SWA was higher in the slow wave sleep (SWS) episodes preceding the episode of parasomnia than in the episodes preceding an awakening without an episode of parasomnia. The temporal course of SWA showed a slower exponential decay in both groups, but the time constant of the curve was larger in parasomniacs than in controls. Finally, in control subjects the sleep spindle index increased from the beginning to the end of the night while it was equally distributed in parasomniacs. CONCLUSIONS: An abnormal deep sleep associated with a high SWS fragmentation might be responsible for the occurrence of SW or ST episodes. PMID- 10802467 TI - Alpha activity and cardiac correlates: three types of relationships during nocturnal sleep. AB - OBJECTIVE: We examined simultaneously alpha activity and cardiac changes during nocturnal sleep, in order to differentiate non-rapid eye movement (NREM) sleep, REM sleep, and intra-sleep awakening. METHODS: Ten male subjects displaying occasionally spontaneous intra-sleep awakenings underwent EEG and cardiac recordings during one experimental night. The heart rate and heart rate variability were calculated over 5 min periods. Heart rate variability was estimated: (1) by the ratio of low frequency (LF) to high frequency (HF) power calculated from spectral analysis of R-R intervals; and (2) by the interbeat autocorrelation coefficient of R-R intervals (rRR). EEG spectral analysis was performed using a fast Fourier transform algorithm. RESULTS: Three types of relationships between alpha waves (8-13 Hz) and cardiac correlates could be distinguished. During NREM sleep, alpha activity and cardiac correlates showed opposite variations, with high levels of alpha power associated with decreased heart rate, rRR and LF/HF ratio, indicating low sympathetic activity. Conversely, during REM sleep, alpha activity was low whereas heart rate, rRR, and the LF/HF ratio peaked, indicating high sympathetic activity. During intra-sleep awakenings, alpha activity and cardiac correlates both increased. No difference in time-course between alpha 1 (8-10 Hz) and alpha 2 (10-13 Hz) activity could be shown. Alpha waves occurred in fronto-central areas during slow wave sleep (SWS), migrated to posterior areas during REM sleep, and were localized in occipital areas during intra-sleep awakenings. CONCLUSIONS: These results suggest that alpha waves are not simply a sign of arousal, as is commonly thought. Fronto central alpha waves, associated with decreased heart rate, possibly reflect sleep maintaining processes. PMID- 10802469 TI - The pain control infusion pump for postoperative pain control in shoulder surgery. AB - PURPOSE: This study was initiated to evaluate the effect of a pain control infusion catheter in managing postoperative pain. TYPE OF STUDY: In a prospective, randomized trial, 62 consecutive patients undergoing arthroscopic subacromial decompression had an indwelling pain control infusion catheter placed at the operative site. MATERIALS AND METHODS: Thirty-one patients received 0.25% bupivacaine and 31 patients received saline infusions, each at a constant rate of 2 mL per hour. Patients evaluated their pain by visual analog scale, and also tabulated the amount of narcotic and nonnarcotic medication used each day in the first week of surgery. RESULTS: There was a statistically significant difference in pain in all parameters tested in the bupivacaine group as compared with the saline control group (P <.05). CONCLUSIONS: The bupivacaine pain control infusion pump is an effective means of decreasing postoperative pain. PMID- 10802470 TI - Meniscal allograft transplantation: a 1- to 5-year follow-up of 22 patients. AB - SUMMARY: Between October 1991 and December 1995, a total of 22 patients (23 knees) underwent meniscal transplantation with nonirradiated, cryopreserved allografts. Implantation was arthroscopically assisted using bone plugs to prevent meniscal extrusion and maintain weight-bearing functional position. Patients were evaluated with an average follow-up of 40 months (range, 13 to 69 months) using the International Knee Documentation Committee, Lysholm, and Tegner scoring systems. The most significant finding was pain reduction after implantation. Lateral and standing anteroposterior radiographs were obtained on all patients at follow-up with an average joint space loss of 0.882 mm (range, 0 to 3 mm). Magnetic resonance imaging was preformed on both knees in 12 of the 22 patients at an average of 24.4 months postoperatively. On average, the allograft meniscus was 63% (range, 31% to 100%) the size of the normal meniscus. Clinical results showed improvement of preoperative pain in all patients. Although patients continue to have good pain relief following their meniscal allograft transplantation, the average shrinkage in the size of the meniscus as shown on magnetic resonance imaging is a concern. PMID- 10802471 TI - Temperature changes associated with radiofrequency energy-induced heating of bovine capsular tissue: evaluation of bipolar RF electrodes. AB - SUMMARY: This in vitro study determined the temperature changes associated with radiofrequency (RF) energy-induced heating of bovine capsular tissue using bipolar RF electrodes. Tissue samples were placed in a saline bath (37 degrees C) and RF energy was applied using 2 different types of bipolar electrodes (VAPR T End Effect and Vapor T Side Effect; Mitek, Westwood, MA). Each electrode was activated for 3 seconds at 10 W, 16 W, and 20 W, for 6 separate data acquisitions. Fluoroptic thermometry designed to be unperturbed by RF fields was used to record temperatures on the tissue surface and at depths of 2 mm, 4 mm, and 5 mm, at 1-second intervals before (5 seconds), during (3 seconds), and after (7 seconds) the application of RF energy. The highest mean temperatures were recorded at the tissue surfaces for the different power settings for each RF electrode type, as follows: End Effect: 48.9 degrees C (10 W), 57.0 degrees C (16 W), and 67.3 degrees C (20 W). Side Effect: 51.5 degrees C (10 W), 62.1 degrees C (16 W), and 71.2 degrees C (20 W). All recorded surface temperatures were within the range known to be acceptable for tissue shrinkage. Gradient effects (i.e., higher-to-lower) were observed for the tissue temperatures measured at the different depth positions. None of the temperatures recorded at the different depths were excessive, suggesting that sensitive anatomic structures should not be damaged by RF energy-induced heating under the conditions described above. PMID- 10802472 TI - Arthroscopic labrum refixation for post-traumatic anterior shoulder instability: suture anchor versus transglenoid fixation technique. AB - SUMMARY: The aim of this retrospective study was to compare recurrence rates following transglenoid labrum refixation or fixation using the suture anchor (FASTak, Arthrex, Naples, FL) technique. Additionally, parameters that apparently influence the rate of redislocation were investigated. There were 163 patients with post-traumatic anterior shoulder instability treated with an arthroscopic labrum refixation; 108 patients (66.3%) were stabilized with the transglenoid suture technique (group I) and 55 patients (33. 7%) with the suture anchor (FASTak) technique (group II). The average follow-up was 4.5 years (range, 2.0 to 7.9 years) in group I and 3.2 years (range, 2.0 to 5.0 years) in group II. The Rowe score increased from a preoperative average of 35.0 points in group I and 35.4 points in group II to a postoperative average of 68.3 points in group I and 84.6 points in group II (P <.01). There was recurrence in 35 patients (32.4%) in group I and 9 patients (16.4%) in group II (P <.05). All incidents of redislocation occurred during the first 21 postoperative months; 58.4% of the patients (n = 63) in group I and 16.4% of the patients in group II (n = 9) had to reduce their sporting activity (P <.001). Independent of the type of surgery, there was a significant correlation of the postoperative rate of redislocation and age (P <.001), number of preoperative dislocations (P <.01), and degree of labrum lesion (P <.001). No correlation with the rate of redislocation was shown for gender, handedness, dislocation-operation interval, degree of Hill-Sachs lesion, or number of transglenoid sutures or anchors. Concerning post-traumatic anterior shoulder instability, the arthroscopic labrum reconstruction with the suture anchor (FASTak) technique was superior to the transglenoid technique but has not yet achieved the level of success obtained by open surgery. With fewer than 5 preoperative redislocations after a first traumatic shoulder dislocation, the arthroscopic treatment is recommended. In cases of more frequent preoperative dislocations, open surgery in combination with a capsular shift should be performed. PMID- 10802473 TI - Arthroscopic assessment of full-thickness rotator cuff tears. AB - SUMMARY: To evaluate the reliability of the arthroscopic assessment of full thickness rotator cuff tears, 117 cases were prospectively investigated by imaging, arthroscopy, and open surgery. The confidence of the surgeon, his accuracy, and the surgeon-dependent character of arthroscopic assessment were evaluated in terms of the description of the main anatomic parameters. The surgeons were confident and accurate in diagnosing a full-thickness tear of the supraspinatus, but they underestimated its coronal and sagittal extent and its reducibility to the greater tuberosity. Conversely, the technique appeared very accurate in describing the rotator interval. Endoscopic assessment was particularly operator-dependent in the anteroposterior analysis of the tear. This study shows the limits of endoscopic assessment of full-thickness rotator cuff tears. It illustrates the need for an adequate arthroscopic technique with a thorough knowledge of normal and pathological anatomy of the rotator cuff. PMID- 10802474 TI - Contaminated anterior cruciate ligament grafts: the efficacy of 3 sterilization agents. AB - PURPOSE: A study was undertaken to determine the incidence of positive cultures resulting from an anterior cruciate ligament (ACL) specimen dropped on the operating room floor and the efficacy of sterilizing the specimen by soaking in 1 of 3 antimicrobial solutions: an antibiotic solution of neomycin and polymyxin B, 10% providone-iodine solution, and standard chlorhexidine gluconate solution. TYPE OF STUDY: Randomized trial. MATERIALS AND METHODS: Fifty ACL specimens removed from patients undergoing total knee arthroplasty were used as the test group. The specimens were longitudinally sectioned into 4 equal pieces. The 4 pieces were dropped on the floor and left for a period of 15 seconds. Cultures were taken from each specimen after immersion in 1 of the 3 sterilization solutions for a period of 90 seconds. One of the 4 specimens was cultured without being exposed to any solution, thereby establishing these specimens as the control group. Cultures of a floor swab were taken at the same time and place that the ACL was dropped. RESULTS: The floor swab cultures were positive in 48 of the 50 specimens (96%). The ACL control group (untreated dropped grafts) had 29 of 50 specimens positive (58%). The grafts soaked in antibiotic solution had 3 of 50 specimens positive (6%). The grafts soaked in providone-iodine solution had 12 of 50 specimens positive (24%). The grafts soaked in chlorhexidine gluconate solution had 1 of 50 specimens positive (in broth only) (2%). CONCLUSION: This study shows that significant contamination occurs when dropping specimens on the floor, as 58% of the dropped grafts had positive cultures. Of the 3 sterilization techniques used, chlorhexidine gluconate seems to be the most efficient with only a single broth culture (2%) found to be positive. The antibiotic solution was second best (6%), although there is no statistically significant difference between these 2 groups. The 10% providone-iodine solution under these test conditions was the least effective of all the 3 sterilization agents with 24% cultures positive after immersion. PMID- 10802475 TI - A simple modified arthroscopic procedure for fixation of displaced tibial eminence fractures. AB - SUMMARY: Avulsion fractures of the tibial eminence have been well described in children and adults with an increased in incidence resulting from road traffic and athletic accidents. According to the literature, only surgical treatment is advocated because of the high incidence of the nonunion and instability following conservative treatment. Open reduction can cause some morbidity and, therefore, arthroscopic techniques have been developed. The limitations of the techniques are related to technical difficulty and unstable fixation. We report a simple and effective method of fixation with the following advantages: (1) use of metallic suture for stable fixation, (2) the fixation device is easily removed in the office, (3) there is no need of additional instruments, and (4) the minimal size of the fixation device avoids damage to the anterior cruciate ligament insertion. This technique has been used in a series of 10 consecutive patients treated for avulsion fractures of the tibial spine since 1991, resulting in a high rate of excellent and good results without any case of nonunion fracture or related complication. PMID- 10802476 TI - Time-dependent reduction in load to failure of wedge-type polyglyconate suture anchors. AB - SUMMARY: Thirty-two absorbable (polyglyconate) and 24 nonabsorbable (polyacetal) wedge-type suture anchors (TAG; Acufex, Mansfield, MA) were implanted into sheep tibiae. Load to failure tests were performed on the day of insertion and at weeks 3, 6, and 12, followed by macroscopic examination. Failure type was suture breakage for nonabsorbable anchors in all groups, with average forces of 142.5 +/ 4.8 N on the first day, 138.0 +/- 6.6 N in week 6, and 135.6 +/- 2.9 N in week 12. In the absorbable group, suture breakage occurred on the first day with a mean force of 133.5 +/- 4. 2 N. In weeks 3, 6, and 12, suture cutout occurred with average forces of 33.75 +/- 5.0 N, 23.25 +/- 2.2 N, and 23.25 +/- 5.9 N, respectively. For absorbable anchors, results at weeks 3, 6, and 12 were significantly lower compared with initial results (P <.001). These results show that wedge-type polyglyconate anchors lose 75% of their initial pullout strength within the first 3 weeks and 84% in 6 weeks. PMID- 10802477 TI - Anterior cruciate ligament reconstruction in the skeletally immature high performance athlete: what to do and when to do it? PMID- 10802478 TI - An intra-articular bioabsorbable interference screw mimicking an acute meniscal tear 8 months after an anterior cruciate ligament reconstruction. AB - SUMMARY: Seven months after a quadrupled semitendinosus anterior cruciate ligament reconstruction, a 44-year-old active-duty soldier reported symptoms consistent with a medial meniscus tear. Preoperative magnetic resonance imaging showed an intra-articular bioabsorbable interference screw within his intercondylar notch. The screw was retrieved arthroscopically. The graft was intact and functional except for a small portion of the anterior fibers, which were debrided. The patient returned to full activities without complaints. PMID- 10802479 TI - The use of the supraclavicular fossa portal in arthroscopic rotator cuff repair. AB - SUMMARY: Advances in arthroscopic technology allow rotator cuff repair through a minimally invasive approach. However, fixation of the rotator cuff tendon to suture anchors can be tedious and time consuming. The supraclavicular fossa portal allows improved access to the tear for passing suture. The authors describe the relevant anatomy, positioning, and surgical technique for use of the supraclavicular fossa portal to simplify arthroscopic rotator cuff repair. PMID- 10802481 TI - Arthroscopy association of north america research award winners PMID- 10802482 TI - Influence of intrauterine environment on development of insulin resistance. PMID- 10802483 TI - Determining the etiology of developmental delay in very young children: what if we had a common internationally accepted protocol?! PMID- 10802484 TI - Mercury, infant neurodevelopment, and vaccination. PMID- 10802485 TI - TT virus infection: an emerging pathogen in search of its identity. PMID- 10802486 TI - By any other name. PMID- 10802487 TI - Ambulatory blood pressure monitoring in the pediatric patient. PMID- 10802488 TI - Correlations between the intrauterine metabolic environment and blood pressure in adolescent offspring of diabetic mothers. AB - OBJECTIVE: To investigate associations between maternal diabetes and blood pressure (BP), obesity, impaired glucose tolerance, and serum lipids in offspring and whether these parameters correlate with metabolism during pregnancy. STUDY DESIGN: Body mass index, BP, serum glucose, and insulin during an oral glucose tolerance test, and lipid concentrations were measured in 99 offspring of diabetic mothers (ODM) and 80 members of a control group. RESULTS: ODM were more obese (body mass index 22.5 +/- 5.6 vs 20.3 +/- 4.0 kg/m(2)) and had higher systolic (8 mm Hg) and mean arterial BP (4 mm Hg) but similar diastolic BP compared with the control group. ODM had higher 2-hour glucose (6.6 +/- 1.3 vs 5.7 +/- 0.9 mmol/L) and insulin (580 +/- 544 vs 377 +/- 239 pmol/L) concentrations but lower fasting concentrations of low-density lipoprotein (2.54 +/- 0.67 vs 2.82 +/- 0.70 mmol/L) and total cholesterol (4.01 +/- 0.80 vs 4.40 +/ 0.78 mmol/L). In both groups body mass index, triglycerides, and fasting and 2 hour glucose concentrations showed correlations with BP measurements. Fasting insulin was correlated with BP readings only in the ODM. Correlations were found between second- and third-trimester maternal free fatty acid concentrations and diastolic and mean arterial BP. Third-trimester beta-hydroxybutyrate was correlated with mean arterial BP. CONCLUSIONS: In ODM, abnormalities in weight and glucose tolerance are associated with abnormal maternal metabolism. Higher BP is an additional abnormality associated with fetal overnutrition. PMID- 10802489 TI - Etiologic yield of subspecialists' evaluation of young children with global developmental delay. AB - OBJECTIVE: To determine the etiologic yield of subspecialists' evaluation of young children with global developmental delay. In addition, variables that may predict finding an underlying etiology were also identified. METHODS: All children <5 years of age, referred over an 18-month period to subspecialty services for initial evaluation of a suspected developmental delay, were prospectively enrolled. Diagnostic yield was ascertained after the completion of clinical assessments and laboratory investigations requested by the evaluating physician. RESULTS: Ninety-nine children (71 boys) were found to have global developmental delay; 96% had a mild or moderate delay documented. An etiologic diagnosis was determined in 44. Four diagnoses (cerebral dysgenesis, hypoxic ischemic encephalopathy, toxin exposure, chromosomal abnormalities) accounted for 34 of 44 (77%) of the diagnoses made. The presence of co-existing autistic traits was associated with significantly decreased diagnostic yield (0/19 vs 44/80, P <.0001), whereas specific historical features (eg, family history, toxin exposure, and perinatal difficulty; 23/32 vs 21/67, P =.0002) and findings on physical examination (eg, dysmorphology, microcephaly, and focal motor findings; 35/48 vs 9/51, P <.0001) were significantly associated with identifying a diagnosis. Multiple logistic regression analysis identified antenatal toxin exposure, microcephaly, focal motor findings, and the absence of autistic traits as significant predictor variables for the identification of an etiology. CONCLUSION: An etiologic diagnosis is often possible in the young child with global developmental delay, particularly in the absence of autistic features. Etiologic yield is augmented by presence of specific findings on history or physical examination on initial assessment. PMID- 10802491 TI - TT virus infection in healthy children and in children with chronic hepatitis B or C. AB - BACKGROUND: The existence of a novel human virus, designated TT virus (TTV), has been reported in association with acute and chronic liver disease of unknown cause. OBJECTIVES: To evaluate the prevalence and clinical significance of TTV infection in childhood. STUDY DESIGN: Sera from 104 healthy children, 85 patients with chronic hepatitis B (HBV) and 29 with chronic hepatitis C (HCV), were tested by polymerase chain reaction with primers corresponding to conserved regions within a part of ORF 2. To characterize polymerase chain reaction products, TTV isolates from 15 subjects were directly sequenced. RESULTS: TTV DNA was detected in 22 (21%) of 104 healthy children and in 55 (65%) of 85 and 20 (69%) of 29 HBV- and HCV-infected individuals, respectively. No significant difference was observed in aminotransferase levels of HBV- or HCV-infected patients with or without TTV coinfection. Sequence analysis showed a high degree of genetic heterogeneity between TTV isolates. CONCLUSIONS: A striking difference was found in the prevalence of TTV between healthy children and patients with chronic HBV or HCV infection (P <.0005). However, based on these results, TTV alone or as coinfection does not seem to cause or exacerbate liver damage in childhood. PMID- 10802490 TI - Maternal seafood diet, methylmercury exposure, and neonatal neurologic function. AB - OBJECTIVE: To determine whether neonatal neurologic function is adversely affected by seafood contaminants from maternal diet during pregnancy. STUDY DESIGN: One hundred eighty-two singleton term births were evaluated in the Faeroe Islands, where marine food includes pilot whale. Maternal serum, hair, and milk and umbilical cord blood were analyzed for contaminants. Levels of essential fatty acids, selenium, and thyroid hormones were determined in cord blood. Each infant's neurologic optimality score was determined at 2 weeks of age adjusted for gestational age, and predictors were assessed by regression analysis. RESULTS: Exposures to methylmercury and polychlorinated biphenyls were increased in relation to maternal seafood intake, as were omega3 fatty acid concentrations in cord serum. Thyroid function was normal. After adjustment for confounders, a 10-fold increase of the cord-blood mercury concentration was associated with a decreased neurologic optimality score of 2.0 (P =. 03). This effect corresponds to a decrease in gestational age of about 3 weeks. Other indicators of the seafood diet had no effect on this outcome. CONCLUSIONS: Prenatal exposure to methylmercury from contaminated seafood was associated with an increased risk of neurodevelopmental deficit. Thus in this North Atlantic population, methylmercury constituted an important neurologic risk factor, although effects of other seafood components were not detectable. PMID- 10802492 TI - Inhaled nitric oxide in term and near-term infants: neurodevelopmental follow-up of the neonatal inhaled nitric oxide study group (NINOS). AB - BACKGROUND: Inhaled nitric oxide (INO) improved oxygenation and reduced the occurrence of death or extracorporeal membrane oxygenation in term and near-term hypoxic neonates. We report the results of neurodevelopmental follow-up of infants enrolled in the NINOS trial. METHODS: Hypoxic infants >/=34 weeks' gestation and <14 days of age were randomized to 20 ppm INO or 100% oxygen as control. Comprehensive neurodevelopmental assessment of survivors occurred at 18 to 24 months of age. RESULTS: A total of 235 infants were enrolled in the original trial. There were 36 deaths, 20 of 121 infants in the control group and 16 of 114 infants in the INO-treated group. Of the 199 surviving infants, 173 (86.9%) were seen for follow-up (88 members of the control group and 85 members of the INO-treated group), and 135 infants were normal (69 [79.3%] members of the control group and 66 [77.6%] members of the INO-treated group). Twenty-two infants had sensorineural hearing loss (12 members of the control group and 10 members of the INO-treated group). Moderate to severe cerebral palsy occurred in 13 infants (7 infants in the control group and 6 infants in the INO-treated group). Mental developmental index scores (87 +/- 18.7 in the control group vs 85 +/- 21.7 in the INO-treated group) and psychomotor developmental index scores (93.6 +/- 17.5 in the control group vs 85.7 +/- 21.2 in the INO-treated group) were not different. A total of 29.6% of the control group compared with 34.5% of the INO-treated group had at least one disability. Infants with congenital diaphragmatic hernia, enrolled in a separate but parallel trial, had similar outcomes with a higher incidence of sensorineural hearing loss. CONCLUSION: Inhaled nitric oxide is not associated with an increase in neurodevelopmental, behavioral, or medical abnormalities at 2 years of age. PMID- 10802493 TI - Effects of in vitro fertilization on low birth weight, preterm delivery, and multiple birth. AB - PURPOSE: To quantify the contribution of in vitro fertilization (IVF) on changes in the rates of low birth weight (LBW), preterm delivery, very low birth weight, and multiple births during the past 3 years. METHODS: Data on IVF pregnancies from 1994 to 1996 within Alberta were reviewed. Population data were obtained from the Provincial notice of a live or stillbirth. RESULTS: The IVF component of increased LBW rate in the province was 17.8% for infants <2500 g and 43.5% for those born <1500 g. IVF accounted for 10.5% of the provincial rate increase in deliveries <37 weeks' gestation and 66.2% of those <30 weeks' gestation. IVF accounted for 21.4% of the twins and all of the sets of triplets in the province. CONCLUSION: During a 3-year period IVF has affected the incidence of LBW, preterm delivery, and multiple birth. IVF is a substantial contributor to changes in very low birth weight and delivery before 30 weeks, which is partly related to multiple births. PMID- 10802494 TI - Continuous pH and Pco2 monitoring during respiratory failure in children with the Paratrend 7 inserted into the peripheral venous system. AB - CONTEXT: The Paratrend monitor provides continuous arterial blood gas monitoring after insertion through a >/=20-gauge arterial cannula. OBJECTIVE: To determine the correlation of arterial blood gas values and the Paratrend monitor placed through a peripheral intravenous catheter. DESIGN: Prospective, open-label evaluation. SETTING: University-based pediatric intensive care unit. PATIENTS: Infants and children with respiratory failure and arterial access. RESULTS: The cohort included 23 infants and children. A total of 100 sample sets (Paratrend/ABG Pco(2) and pH values) were collected. The absolute difference between the arterial and Paratrend Pco(2) was 2. 9 +/- 1.8 mm Hg (range 0 to 9 mm Hg). Linear regression analysis of Paratrend Pco(2) versus arterial Pco(2) resulted in r = 0.97 and r(2) = 0.9479 (P <.001). Bland-Altman analysis of Pco(2) values demonstrated a bias +/- precision of -2.1 +/- 2.7 mm Hg. The absolute difference between arterial and Paratrend pH was 0.04 +/- 0. 02 units (range 0 to 0.15 units). Linear regression analysis of Paratrend pH versus arterial pH resulted in r = 0.83 and r(2) = 0. 7016 (P <.0001). Bland-Altman analysis of pH values revealed a bias +/- precision of 0.03 +/- 0.03 units. CONCLUSIONS: Inserted through a peripheral intravenous cannula, the Paratrend monitor can be used to provide an accurate estimation of arterial blood gas values in children with respiratory failure. PMID- 10802495 TI - Increased urinary peroxides in newborn infants receiving parenteral nutrition exposed to light. AB - OBJECTIVES: To determine whether peroxide loads infused with total parenteral nutrition (TPN) are fully quenched by premature infants. STUDY DESIGN: After baseline urine peroxide levels were established, the effect of various parenteral regimens was correlated with urinary peroxide levels in 64 newborn infants /=2500 g). DESIGN/METHODS: Cross-sectional design. Fifty-three ELBW and 53 NBW adolescents without a major neurodevelopmental disability were matched by sex, race, age, and socioeconomic status. Anthropometrics (z scores), bone age, body composition (Lunar DPX-L densitometry), and sexual maturity were assessed. ELBW adolescents were classified as being born small for gestational age (SGA) or not (NSGA). RESULTS: Subjects were 58.5% female, 43.4% black, and 56.6% white. The mean birth weight for ELBW subjects was 849 g and 3355 g for NBW subjects. The mean age was 14.85 years. On average, ELBW adolescents were 4.8 cm shorter and 9.1 kg lighter than NBW adolescents. ELBW adolescents had lower mean z scores for height (P <.0001), weight (P <.0001), and head circumference (P <.0001) than NBW adolescents. ELBW/SGA subjects had lower mean z scores for height (P <.0001) and weight (P =.001) than NBW subjects. Head circumference z scores were lower for the ELBW/SGA group than the ELBW/NSGA group or the NBW group (P =. 003). Sexual maturity and relative body composition were similar between groups. Bone age, measured in SD units, was more advanced in the ELBW group (0.86 vs. 0.42, P =.039). CONCLUSIONS: ELBW adolescents who survive without a major neurodevelopmental disability attain lower growth measurements compared with NBW adolescents but have similar sexual maturation and relative body composition. PMID- 10802497 TI - Role of food protein intolerance in infants with persistent distress attributed to reflux esophagitis. AB - BACKGROUND: Distressed behavior is common in infants and is often attributed to gastroesophageal reflux (GER) or food protein intolerance. OBJECTIVE: To examine the effect of a hypoallergenic amino acid-based infant formula (AAF) on distressed behavior and GER symptoms in infants who failed to respond to extensively hydrolyzed formula and antireflux medications. STUDY DESIGN: Nineteen distressed infants (9 boys and 10 girls; median age, 5.0 months) with presumed GER underwent gastroscopy (n = 17) and esophageal 24-hour pH monitoring (n = 14). Double-blind placebo-controlled (DBPC) formula challenges of AAF versus previously besttolerated formula were conducted. RESULTS: Nine infants had histologic evidence of esophagitis, and 9 had inflammatory changes in the stomach and/or duodenum. Symptoms remitted in all infants within 2 weeks of the start of feeding with AAF. On DBPC challenge after a median period of 3 months of receiving AAF, 12 infants were intolerant to active formula (distress score, 287 vs 580 min/wk,P =. 01; symptom score, 23.1 vs 36.1, P =.03). Seven infants did not relapse and were considered tolerant (distress score, 470 vs 581, P =.77; symptom score, 29.5 vs 20.2; P =.89). CONCLUSION: Treatment with AAF may reduce distressed behavior and symptoms of GER in infants with food protein intolerance. PMID- 10802498 TI - Bone mineral status in prepubertal children with cystic fibrosis. AB - OBJECTIVES: To determine whether osteopenia is evident in prepubertal children with cystic fibrosis (CF) and, if so, whether it is caused by a deficiency in bone formation or increased bone resorption. STUDY DESIGN: With the use of a prospective case control study design, we investigated 11 prepubertal children with CF between the ages of 8 and 12 years old and a non-CF control group matched by weight and sex. Bone density at the radius, ulnar, trochanter, femoral neck, and lumbar spine, biochemical markers of bone metabolism, calcium, vitamin D metabolites, and intact parathyroid hormone were measured in all subjects. Comparisons between the 2 groups were performed with Wilcoxon matched pairs and Fisher exact tests. RESULTS: Intake of total calories, calcium, phosphorus, and vitamin D was significantly greater in the CF group than in the control group. Serum 25(OH)vitamin D levels were significantly lower in the CF group: median 22 ng/mL for the CF group and 39 ng/mL for the control group (P =.02). 1,25(OH)(2) vitamin D levels were borderline or low in 7 subjects in the CF group and 2 members of the control group (P =.08, Fisher exact test). Intact parathyroid hormone levels were higher than the upper limit of normal in 4 subjects of the CF group and 1 member of the control group. Despite these biochemical abnormalities, we found no evidence of bone mineral deficiency in the CF group. CONCLUSIONS: Prepubertal children with CF do not have bone mineral deficit compared with a weight- and sex-matched control group; however, their lower vitamin D levels may portend problems with bone mineralization during adolescence and adulthood. PMID- 10802499 TI - Etiology and management of pediatric chylothorax. AB - OBJECTIVES: To determine the incidence and etiology of chylothorax and to assess our therapeutic management approach. STUDY DESIGN: We reviewed 51 patients diagnosed with chylothorax over a 12-year period. Cause, interval between operation and diagnosis, duration of chylothorax, and total volume loss per weight were recorded. RESULTS: Chylothorax was diagnosed in 46 children after cardiothoracic surgery, giving an incidence of 2.5% (46/1842); in 1 child chylothorax occurred after chest trauma, and in 4 the chylothorax was congenital or a manifestation of lymph angiomatosis. Three etiologic groups were identified: group 1, direct injury to the thoracic duct (33/51 = 65%); group 2, thrombosis and/or high venous pressure in the superior vena cava (14/51 = 27%); and group 3, congenital (4/51 = 8%). Conservative treatment was the only treatment in 80% of the patients. Surgical procedures consisted of 4 ligations of the thoracic duct, placement of 7 pleurodesis shunts, and placement of 2 pleuroperitoneal shunts. Patients in groups 2 and 3 were at higher risk for failure of conservative treatment (P <. 005). Longer duration of chylothorax and higher volume of drainage were present in group 2 compared with group 1 (P <.01). CONCLUSION: Conservative treatment was successful in 80% of the patients with our management approach. Prevention, early recognition, and treatment of potential complications, such as superior vena cava thrombosis or obstruction, may further improve success of conservative treatment. Congenital chylothorax seems different and may require a specific approach. PMID- 10802500 TI - Resting energy expenditure in disorders of propionate metabolism. AB - OBJECTIVES: During intercurrent illness children with methylmalonic acidemia were found to have increased resting energy expenditure (REE). We measured REE in children with disorders of propionate metabolism (methylmalonic and propionic acidemia) when they were well and compared the values with those predicted by the Schofield equation. STUDY DESIGN: Prospective study in tertiary care facility. REE was measured with open-circuit indirect calorimetry under standardized conditions. Predicted REE values were calculated with the Schofield equation. Fourteen subjects with propionic acidemia (n = 3) and methylmalonic acidemia (n = 11) were studied. RESULTS: The median REE was 690 kcal/d (range 186 to 1687 kcal/d), which is significantly reduced, representing 80% +/- 18% of that predicted by the Schofield height and weight equation (P <.01). REE was significantly lower in female compared with male patients for unknown reasons. There were no differences with age or neurologic state. REE was not further reduced in those with chronic renal failure. CONCLUSION: REE in patients with disorders of propionate metabolism is reduced when they are well. PMID- 10802501 TI - Type 2 diabetes among North American children and adolescents: an epidemiologic review and a public health perspective. AB - OBJECTIVES: To review the magnitude, characteristics, and public health importance of type 2 diabetes in North American youth. RESULTS: Among 15- to 19 year-old North American Indians, prevalence of type 2 diabetes per 1000 was 50.9 for Pima Indians, 4.5 for all US American Indians, and 2.3 for Canadian Cree and Ojibway Indians in Manitoba. From 1967-1976 to 1987-1996, prevalence increased 6 fold for Pima Indian adolescents. Among African Americans and whites aged 10 to 19 years in Ohio, type 2 diabetes accounted for 33% of all cases of diabetes. Youth with type 2 diabetes were generally 10 to 19 years old, were obese and had a family history of type 2 diabetes, had acanthosis nigricans, belonged to minority populations, and were more likely to be girls than boys. At follow-up, glucose control was often poor, and diabetic complications could occur early. CONCLUSIONS: Type 2 diabetes is an important problem among American Indian and First Nation youth. Other populations have not been well studied, but cases are now occurring in all population groups, especially in ethnic minorities. Type 2 diabetes among youth is an emerging public health problem, for which there is a great potential to improve primary and secondary prevention. PMID- 10802502 TI - Talking with families about herbal therapies. PMID- 10802503 TI - Iatrogenic exposure to mercury after hepatitis B vaccination in preterm infants. AB - Thimerosal, a derivative of mercury, is used as a preservative in hepatitis B vaccines. We measured total mercury levels before and after the administration of this vaccine in 15 preterm and 5 term infants. Comparison of pre- and post vaccination mercury levels showed a significant increase in both preterm and term infants after vaccination. Additionally, post-vaccination mercury levels were significantly higher in preterm infants as compared with term infants. Because mercury is known to be a potential neurotoxin to infants, further study of its pharmacodynamics is warranted. PMID- 10802504 TI - Response to steroid therapy in autism secondary to autoimmune lymphoproliferative syndrome. AB - We report a child who developed autoimmune lymphoproliferative syndrome (ALPS) secondary to a heterozygous dominant negative mutation in the death domain of the Fas receptor. Previously developmentally normal, he had symptoms of autism with rapid regression in developmental milestones coincident with the onset of lymphoproliferation and autoimmune hemolytic anemia. Low-dose steroid therapy induced early and complete remission in the ALPS phenotype. There was subjective improvement, followed by objective improvement in speech and developmental milestones. We propose that autism may be part of the autoimmune disease spectrum of ALPS in this child, and this case represents a novel manifestation and target organ involvement in this disease. PMID- 10802505 TI - Zinc-induced anemia and neutropenia in an adolescent. AB - We report an adolescent who developed anemia, leukopenia, and neutropenia after prolonged use of over-the-counter zinc for treatment of acne. Hypocupremia and sideroblastic anemia may result from long-term or excessive exposure to zinc. PMID- 10802506 TI - Budd-Chiari syndrome associated with coagulation abnormalities in a child with carbohydrate deficient glycoprotein syndrome type Ix. AB - A 6-year-old male patient presented with Budd-Chiari syndrome and glycoprotein abnormalities associated with carbohydrate deficient glycoprotein syndrome type I with yet unidentified molecular defect (type Ix). Budd-Chiari syndrome most likely developed after hepatic venous thrombosis caused by coagulation abnormalities resulting from hypoglycosylation and functional impairment of anticoagulant proteins. PMID- 10802507 TI - Inguinal hernias containing the uterus, fallopian tube, and ovary in premature female infants. AB - Inguinal hernias were diagnosed at 42 and 38 weeks' postconceptional age in 2 premature girls. The hernial sac contained the uterus, one Fallopian tube, and one ovary. The diagnosis was made by physical and sonographic examination and was confirmed during surgical correction. We suggest sonography in the diagnostic workup in (premature) female infants with an inguinal hernia. PMID- 10802508 TI - The pharmacologic mechanism by which inhaled epinephrine reduces airway obstruction in respiratory syncytial virus-associated bronchiolitis. AB - Inhaled racemic epinephrine relieves airway obstruction in patients with respiratory syncytial virus bronchiolitis. The contribution of alpha- versus beta adrenoreceptor stimulation toward this clinical effect is unknown. We describe an infant treated with propranolol for supraventricular tachycardia in whom respiratory syncytial virus bronchiolitis developed. Inhaled racemic epinephrine improved his respiratory symptoms, whereas nebulized albuterol did not. PMID- 10802509 TI - Use of plastic-coated stylets for endotracheal intubation. PMID- 10802511 TI - Reply PMID- 10802510 TI - When should hyperglycemia be treated in cystic fibrosis? PMID- 10802513 TI - Reply PMID- 10802512 TI - In partial defense of prolonged indomethacin therapy for patent ductus arteriosus. PMID- 10802515 TI - Reply PMID- 10802514 TI - Intracranial hemorrhages in Alagille syndrome. PMID- 10802516 TI - Ghrelin: a step forward in the understanding of somatotroph cell function and growth regulation. PMID- 10802517 TI - The role of Y chromosome deletions in male infertility. AB - Male infertility affects approximately 2-7% of couples around the world. Over one in ten men who seek help at infertility clinics are diagnosed as severely oligospermic or azoospermic. Recent extensive molecular studies have revealed that deletions in the azoospermia factor region of the long arm of the Y chromosome are associated with severe spermatogenic impairment (absent or severely reduced germ cell development). Genetic research into male infertility, in the last 7 years, has resulted in the isolation of a great number of genes or gene families on the Y chromosome, some of which are believed to influence spermatogenesis. PMID- 10802519 TI - Psychoses associated with thyrotoxicosis - 'thyrotoxic psychosis.' A report of 18 cases, with statistical analysis of incidence. AB - OBJECTIVE: To report a series of newly diagnosed thyrotoxic patients with concurrent acute psychosis, and to assess the association between the two disorders. DESIGN: Retrospective study of thyrotoxic patients with associated psychosis ('thyrotoxic psychosis'; TP) requiring inpatient psychiatric care. New Zealand thyrotoxicosis annual incidence figures and first psychiatric admission rates for affective psychosis were utilised to statistically assess the co occurrence of thyrotoxicosis and affective psychosis. PATIENTS AND METHODS: During the 20-year study period, 18 inpatients (16 women and 2 men), mean age 54 years, with TP were identified. No patient had a past history of thyrotoxicosis, but four had required psychiatric inpatient care many years earlier. Thyrotoxicosis was documented by radioimmunoassay of thyroid hormone levels, and thyroid scintiscan. Psychiatric manifestations were classified using ICD9 criteria. RESULTS: Thyroid hormone levels were markedly elevated in more than half of our TP patients. All younger patients had Graves' disease, and most older patients toxic nodular goitre. All patients were treated with antithyroid drugs, and all but one subsequently received (131)I therapy. Two patients were not mentally ill when thyrotoxicosis was diagnosed, but suffered major mood swings when thyroid hormone levels were falling. There was no specific psychiatric clinical picture but affective psychoses were commonest - seven depression, seven mania. The other diagnoses were two schizophreniform, one paranoid, and one delirium. Initially, neuroleptic medication was used in all but one patient, and during long-term follow-up (median 11 years) more than half our series had remained well with no further psychiatric problems. Statistical analysis was restricted to thyrotoxic patients with first psychiatric hospital admission for affective psychosis. During the 20-year period, there were nine thyrotoxic patients (95% confidence interval 4.5-17.1) with concurrent affective psychosis requiring first admission, and the calculated expected number was only 0.36. These findings indicate a clear association well above chance co-occurrence. CONCLUSION: TP is not a specific clinical picture, but affective psychoses are commonest. Statistical analysis of thyrotoxic patients with concurrent affective psychoses showed an incidence well above chance co-occurrence. It appears that thyrotoxicosis may be a precipitant of acute affective psychosis. PMID- 10802518 TI - Virologic factors related to interferon-alpha-induced thyroid dysfunction in patients with chronic hepatitis C. AB - OBJECTIVE: Hepatitis C virus (HCV), being reported to be associated with a high prevalence of serological markers of autoimmunity in HCV-infected patients, and possibly sharing partial sequences in amino acid segments with thyroid tissue antigens, may be associated with interferon-alpha (IFN-alpha)-induced thyroid dysfunction in chronic hepatitis C patients. We conducted this study to clarify the issue. DESIGN AND METHODS: One hundred and fifty chronic hepatitis C patients with normal baseline thyroid function were treated with IFN-alpha 2a, 2b and n1 (3-6 million Units three times weekly for 24 weeks). Pretreatment sera were tested for HCV genotype and HCV RNA levels. Serum thyrotropin, total thyroxine and free thyroxine index were performed every 4 weeks for 24 weeks followed by every 8 weeks for another 24 weeks. RESULTS: Twenty-one (14.0%) patients developed early thyroid dysfunction (abnormal thyroid function during the first 3 months of therapy). Female gender, lower HCV RNA levels, IFN-alpha n1 and a lower IFN-alpha dose were significantly associated with early thyroid dysfunction. On multivariate analysis, gender, IFN-alpha preparation and HCV RNA levels were the significant factors associated with early thyroid dysfunction. Seven (4.7%) patients developed thyroid dysfunction during the second 3 months of IFN-alpha therapy. Taken together, 18.7% patients developed thyroid dysfunction. Female, mixed HCV genotype infection and lower HCV RNA levels were significantly associated with thyroid dysfunction. However, only gender remained significantly associated with IFN-alpha-induced thyroid dysfunction in multivariate analysis. CONCLUSIONS: The virologic features of HCV may be associated with thyroid dysfunction in chronic hepatitis C patients treated with IFN-alpha. Nevertheless, gender still plays the most important role in IFN-alpha-induced thyroid dysfunction. PMID- 10802520 TI - The effect of long-term, non-suppressive levothyroxine treatment on quantitative ultrasonometry of bone in women. AB - OBJECTIVE: To evaluate the impact of long-term, non-suppressive levothyroxine (L T(4)) treatment on quantitative ultrasonometry in women. DESIGN: This was a case control study. SUBJECTS AND METHODS: Altogether 667 women (mean age+/-s.d., 49.5+/-13.1 years) were studied. Of these, 156 (23%) had non-toxic goitre or hypothyroidism and had been taking L-T(4) (75-100 microg/day) for at least 5 years (mean+/-s.d., 12.5+/-7.5 years); the remaining 511 (77%) women were not receiving L-T(4). All women had completed a questionnaire on risk factors for thyroid dysfunction and osteoporosis, and those with diseases or treatments known to effect bone metabolism - other than thyroxine or hormone replacement therapy (HRT) - were excluded. Women underwent quantitative ultrasonometry (QUS) at the heel. Speed of sound (SOS), broadband ultrasound attenuation (BUA) and the stiffness index (SI) were compared, first, in all women taking L-T(4) and controls and, secondly, in women taking L-T(4) and controls pair-matched for age, weight, body mass index (BMI), menopausal status and HRT use. RESULTS: Even after matching for age, weight, BMI, menopausal and HRT status, women taking L-T(4) had significantly lower values for SOS and SI (P<0.05), but not for BUA. However, absolute T- and Z-scores for SI were not low in either the study or control groups. Lower values were associated, but not significantly so, with years since the menopause and duration of L-T(4) treatment. CONCLUSIONS: Long-term, non suppressive L-T(4) treatment in women with goitre or hypothyroidism was associated with a slight reduction in QUS values, which was more pronounced in postmenopausal women. This group could be at higher risk for osteoporotic fracture. PMID- 10802521 TI - Immediate changes in biochemical markers of bone turnover and circulating interleukin-6 after parathyroidectomy for primary hyperparathyroidism. AB - OBJECTIVE: The time course of the immediate change in bone turnover after parathyroidectomy (PTX) for primary hyperparathyroidism (PHPT) is not clear. It is uncertain whether circulating interleukin-6 (IL-6) plays a role in mediating the acute withdrawal of the effects of parathyroid hormone (PTH) on bone turnover after PTX. The aims of this study were to determine the time course of immediate changes in biochemical markers of bone turnover after PTX and whether circulating IL-6 is involved in the immediate changes of bone turnover after PTX. DESIGN AND METHODS: IL-6 and bone turnover markers were measured in eight women (aged 55+/ 11 years, mean+/-s.d. ) with PHTP at baseline and at 1-2h, and 1, 2, 5, 7 and 12 days after PTX. We compared the results with those from eight individually matched women (healthy controls) and five subjects undergoing major surgery (surgical controls). RESULTS: At baseline, serum levels of IL-6 and bone turnover markers were higher in PHPT than those in healthy controls (P<0.05). Serum levels of procollagen propeptides increased by 22 and 27% at days 2 and 5, respectively, compared with baseline (P<0.05). Serum tartrate-resistant acid phosphatase decreased by 2 days after PTX, and urinary collagen crosslinks decreased significantly by 21-41% within 24h (P<0.05). Serum IL-6 levels increased immediately in both PHPT and surgical controls at postoperative follow-up (repeated measures ANOVA). CONCLUSIONS: (1) PTX decreases bone resorption immediately and (2) circulating IL-6 is not involved in the changes in bone turnover immediately after PTX. PMID- 10802522 TI - Birth weight influences long-term catch-up growth and height prognosis of GH deficient children treated before the age of 2 years. AB - OBJECTIVE: To investigate which pretreatment variables most significantly affect long-term growth response to GH therapy in children with apparently idiopathic GH deficiency (GHD) treated from a similar and very young age (less than 2 years), for the same period (7 years) and with the same therapeutic protocol. DESIGN AND METHODS: Twelve children with either isolated GHD or multiple pituitary hormone deficiency were treated with biosynthetic human GH (0.7IU/kg per week) and were examined every 6 months. Height measurements were performed by Harpenden stadiometers. Bone age was evaluated every 12 months. RESULTS: The onset of therapy was followed in all patients by an important height gain, which attained its zenith during the first year of treatment and became progressively less evident during the next 4 years. Cumulative height gain was 3.0+/-1.7SDS. Thanks to the therapy, at the end of the 7-year treatment period, average height in the entire series was not significantly far from mean target height (TH) (-0.7+/-1.3 vs -0. 3+/-0.4SDS) and average predicted height (PH) (-0.2+/-1.4SDS) was very close to TH. A stepwise regression analysis showed that both catch-up growth under therapy and PH at the end of the 7-year treatment period were positively influenced by birth weight (BW). CONCLUSIONS: a) Our 7-year prospective study on GHD infants treated with GH from less than 2 years of age confirmed the importance of early diagnosis and treatment of GHD in childhood. b) The influence of BW on growth response to GH therapy in GHD children persists over time, at least when treatment is begun from less than 2 years of age. PMID- 10802523 TI - Allopregnanolone and dehydroepiandrosterone response to corticotropin-releasing factor in patients suffering from Alzheimer's disease and vascular dementia. AB - OBJECTIVE: Neurosteroids have been suggested to be involved in the regulation of cognitive performances. A major neurosteroid gamma-aminobutyric acid (GABA) agonist is allopregnanolone: the main source of circulating allopregnanolone is the adrenal cortex. Studies indicated that a disturbance of the central regulation of the hypothalamic-pituitary-adrenocortical axis occurs in both senile (Alzheimer's disease: AD) and vascular dementia (VD). DESIGN: The aim of the present study was to evaluate the levels of circulating allopregnanolone, dehydroepiandrosterone (DHEA) and cortisol and their response to corticotropin releasing factor (CRF) test in AD and VD. METHODS: Three groups of 12 subjects were included in the study: AD, VD and age-matched control subjects. CRF test was performed in all subjects and allopregnanolone, DHEA and cortisol levels were measured every 15min for 2h. RESULTS: Mean +/- s.e.m. allopregnanolone and DHEA basal levels were significantly lower in AD and VD than in controls, while cortisol levels were significantly higher than in controls (P<0.01). Allopregnanolone and DHEA levels increase in response to CRF test in all subjects but the area under curve (AUC) in patients was significantly lower than in controls (P<0.01). Cortisol secretion appeared to be very sensitive in response to CRF stimulation: in fact, cortisol response to CRF test in AD and VD subjects was higher (both as AUC and as % max increase) than in controls (P<0.01). CONCLUSIONS: The present study firstly showed that allopregnanolone levels are reduced both in AD and in VD and that dementia has a preserved stimulated response of allopregnanolone to CRF. Overall, however, the total response of allopregnanolone to CRF remains reduced in respect to controls. Further studies are necessary for a better understanding of the role of neurosteroids in the regulation of cognitive function. PMID- 10802525 TI - Unexplained habitual abortion is associated with a reduced endometrial release of soluble intercellular adhesion molecule-1 in the luteal phase of the cycle. AB - Although the mechanisms causing recurrent spontaneous abortion (RSA) remain frequently speculative, recent evidence indicates that a specific uterine immune endocrine network plays a pivotal role in the continuation of pregnancy. We have recently demonstrated that an adhesion molecule of the immune system, named intercellular adhesion molecule (ICAM)-1, is markedly expressed at both protein and mRNA levels in endometrial stromal cells and is able to mediate their interaction with lymphoid cells. Moreover, we have shown that the soluble form of ICAM-1 (sICAM-1) can be released by the endometrium in a hormone-dependent manner. The present study was designed to determine whether surface and/or sICAM 1 expression by cultured endometrial stromal cells could be related to early pregnancy loss in patients with a history of unexplained RSA. Luteal-phase endometrial biopsies were obtained from eight patients who had experienced three or more consecutive unexplained RSAs in the first trimester and 12 control fertile women. Surface ICAM-1 was similarly expressed on luteal-phase endometrial cells obtained from women with and without a history of unexplained RSA. In contrast, the endometrial release of sICAM-1 was significantly lower in abortion prone patients than in control women. sICAM-1 is a cytokine-inducible molecule able to interfere with several immunological responses and the reduced levels of the protein shed by the endometrium in patients who have suffered from unexplained RSAs may reflect the presence of an altered immunological environment during the early phases of pregnancy. PMID- 10802524 TI - Decreased ligand affinity rather than glucocorticoid receptor down-regulation in patients with endogenous Cushing's syndrome. AB - OBJECTIVE: Glucocorticoids (GCs) serve a variety of important functions throughout the body. The synthesis and secretion of GCs are under the strict influence of the hypothalamo-pituitary-adrenal axis. The mechanisms of action of GCs are mediated by the intracellular glucocorticoid receptor (GR). Over the years, many studies have been performed concerning the regulation of GR expression by GC concentrations. METHODS: In the present study, we determined the characteristics of the GR in peripheral mononuclear blood leukocytes (PBML) from thirteen patients with endogenous Cushing's syndrome and fifteen control subjects, using a whole cell dexamethasone binding assay. Furthermore, cortisol concentrations were determined in order to investigate a possible relationship between serum cortisol levels and receptor characteristics. RESULTS: There were no differences in mean receptor number between patients and controls. On the other hand, a significantly lower ligand affinity was identified in cells from patients with Cushing's syndrome compared with controls. A complete normalisation of the ligand affinity was observed after treatment in the only patient tested in this respect, whereas the receptor number was not affected. In patients, there was a statistically significant negative correlation between cortisol concentrations and ligand affinity, which was not found in controls. CONCLUSION: Receptor down-regulation does not occur in PBML from patients with endogenous Cushing's syndrome. On the other hand, there seems to be a diminished ligand affinity which possibly reflects receptor modification in response to exposure to the continuously high cortisol levels in patients with Cushing's syndrome. This assumption is substantiated by the fact that in one patient a normalisation of the ligand affinity after complete remission of the disease was seen. PMID- 10802526 TI - Secretion of inhibin B by human prepubertal testicular cells in culture. AB - OBJECTIVE: Inhibin B is a secretory product of Sertoli cells of the human testis. It has been reported that serum levels of inhibin B in infant boys, peaking at 3 months of age, exceed levels in adult men. The aim of this study was to evaluate inhibin B secretion in primary prepubertal mixed testicular cell cultures, prepared from testes collected at necropsy. DESIGN AND METHODS: Cell cultures were divided into three age groups on the basis of differences in testicular histology: group 1 (n = 7), 1- to 10-day-old newborns, group 2 (n = 7), 1- to 9 month-old infants, and group 3 (n = 8), 12- to 84-month-old children. Cells were maintained in culture for 6 days, harvested and counted. In some samples, during the last 4 days, cells were stimulated with 10ng/ml highly purified human (h) LH (n = 9), 2 ng/ml recombinant human (rh) FSH (n = 9) or 50 ng/ml rhGH (n = 4). On day 6, the secretion of inhibin B and testosterone into the medium was estimated in triplicate. Inhibin B was determined by ELISA and testosterone by RIA. RESULTS: Median (range) inhibin B secretion was 465 (225-1007), 275 (107-298), and 58 (15-184) pg/million cells.24h in groups 1, 2 and 3 respectively. A logarithmic transformation of these values was performed to normalize data. Mean+/-s.d. of transformed inhibin B secretion in group 1 was significantly higher than in group 2 or 3 (P<0.005) and the values for groups 1 and 2 were significantly higher than that for group 3 (P< 0.005). No significant correlation between testosterone and inhibin B secretion into the medium was found when the 22 culture samples were analyzed as a whole. Inhibin B secretion was significantly increased after stimulation with highly purified hLH, rhFSH and rhGH (P < 0.05) and a significant positive correlation between inhibin B and testosterone was found under both hLH and rhFSH stimulation. CONCLUSIONS: It is concluded that cells collected from newborns have the highest capacity to secrete inhibin B in vitro, and that this capacity decreases with age during the first years of life. Since no data are available on serum inhibin levels in newborns, it is possible that concentrations at 3 months of age do not represent a post natal peak but a declining level of high newborn values. As expected, FSH stimulated inhibin B secretion in culture. LH stimulation was probably mediated by paracrine factors secreted by interstitial cells. Finally, our results add new evidence of the involvement of GH in testicular maturation. PMID- 10802527 TI - Resting metabolic rate, body composition, and serum leptin concentrations in a free-living elderly population. AB - OBJECTIVE: The present study investigated the relationship between serum leptin concentrations and resting metabolic rate (RMR) in a large study group of elderly individuals with special consideration of body composition and body fat distribution as possible confounders. DESIGN AND METHODS: The subjects were 122 women (age: 69+/-6 years, body mass index (BMI): 26.3+/-3.6 kg/m(2)) and 82 men (age: 69+/-5 years, BMI: 26.0+/-2.6 kg/m(2)). RMR was measured by indirect calorimetry and body composition by the bioelectrical impedance method. Serum leptin levels were determined by radioimmunoassay. RESULTS: There was a strong correlation between fat mass (FM) and serum leptin levels in both sexes. An age related decline in leptin levels adjusted for FM was observed only in the women. After adjustment of RMR for both fat-free mass (FFM) and FM, leptin levels were not associated with RMR. In stepwise multiple regression analysis, FFM was the main predictor of RMR, explaining 35.8% and 47.6% of the variance of RMR in men and women respectively. FM did not explain variance in RMR in men, but accounted for 2.6% of the variance in RMR in women. Waist-hip-ratio and age influenced RMR only in males, explaining 5.7% and 4.0% of the variance in RMR respectively. CONCLUSION: Leptin is not a significant predictor of RMR in the elderly, but body composition and distribution of body fat are significantly associated with RMR. PMID- 10802528 TI - Thyrocyte release of asymmetric dimethylarginine does not account for human thyrocyte inhibition of endothelial cell cyclic GMP. AB - BACKGROUND: The thyroid gland produces and responds to the signalling molecule nitric oxide (NO). The activity of NO synthase (NOS) may be regulated by endogenous NOS inhibitors such as asymmetric dimethylarginine (ADMA). OBJECTIVE: To investigate whether human thyrocytes are capable of regulating NOS activity via the production of ADMA. DESIGN: Human thyrocytes were incubated with human umbilical vein endothelial cells (HUVEC) in order to determine the effect on HUVEC NOS activity. HUVEC cGMP production over a 3-h period was measured as an indicator of NOS activity in the absence and presence of thyrocytes. To determine thyrocyte production of ADMA, samples of conditioned media were analysed by HPLC. RESULTS: The presence of primary human thyrocytes or immortalized human thyrocyte SGHTL-189 cells caused a significant inhibition of both basal (approximately 57% inhibition) and thrombin-stimulated (approximately 42% inhibition) HUVEC cGMP production. Both primary human thyrocytes and SGHTL-189 cells released ADMA (approximately 0. 28 microg per 10(6) thyrocytes over a 3-day period). However, excess L-arginine, the natural substrate for NOS, was unable to overcome thyrocyte inhibition of HUVEC cGMP production. CONCLUSION: These data indicate that human thyrocytes potently reduce endothelial cell cGMP concentrations, and that thyrocytes produce the endogenous NOS inhibitor, ADMA. However, the inhibition of endothelial cGMP is not mediated via thyrocyte production of a competitive NOS inhibitor. PMID- 10802529 TI - Calcium stimulates parathyroid hormone-related protein production in Leydig tumor cells through a putative cation-sensing mechanism. AB - The production of parathyroid hormone-related protein (PTHrP) is regulated by a variety of hormones and growth factors. Previous research has shown that several PTHrP-producing cells are influenced by extracellular calcium (Ca(2+)(o)) concentration, with elevated levels increasing PTH-like activity released by cultured H500 rat Leydig tumor cells through a post-transcriptional mechanism. We have investigated the hypothesis that calcium stimulates PTHrP production in H500 cells by interacting with a cell membrane-associated cation-sensing receptor. Besides increased Ca(2+)(o) concentration, magnesium and the polycationic antibiotic neomycin also increased PTHrP production in a concentration-dependent manner. In the presence of the calcium ionophore, ionomycin, which markedly elevated cytosolic free calcium, the stimulation by Ca(2+)(o) of PTHrP could still be detected. These results indicate that increasing Ca(2+)(o) stimulates PTHrP production, possibly through a putative cell membrane-associated calcium sensing mechanism. RT-PCR revealed the presence of a very small amount of calcium sensing receptor coding mRNA. PMID- 10802530 TI - Effect of activin on production and secretion of prolactin and growth hormone in cultured rat GH3 cells. AB - OBJECTIVE: To evaluate the effect of the growth factor activin A on the secretion of prolactin (PRL) and GH in cultured GH3 cells. METHODS: The concentrations of PRL and GH secreted from GH3 cells cultured in media with and without activin A were measured by RIA, and the expression of PRL mRNA and GH mRNA were analyzed using the Northern blot method. RESULTS: Activin A significantly inhibited PRL release from GH3 cells cultured for 48h in a dose-dependent manner (activin: 0.3 3nM). The inhibitory effects of 3nM activin A were observed in the culture from 12h to 48h (53.2% of control). Activin A (3nM) also significantly inhibited the expression of PRL mRNA at 24h (33.8% of control). In contrast, activin A significantly stimulated GH release from GH3 cells cultured for 48h in a dose dependent manner (activin: 0.3-3nM). The stimulatory effect of 3nM activin A was observed in the culture for 48h (157.6% of control). Activin A (3nM) also significantly stimulated the expression of GH mRNA at 24h (183.6% of control). In spite of these significant changes in PRL and GH secretion, pit-1 mRNA levels were not significantly changed by activin A. CONCLUSIONS: These findings indicated that activin A modulates PRL and GH secretion through the regulation of PRL and GH gene transcription in GH3 cells, but that these effects are unrelated to pit-1 gene expression. PMID- 10802531 TI - O-glycosylation delays the clearance of human IGF-binding protein-6 from the circulation. AB - OBJECTIVE: The actions of insulin-like growth factors (IGF-I and IGF-II) are modulated by a family of six structurally related, high-affinity binding proteins (IGFBPs 1-6). IGFBP-6, an O-linked glycoprotein, preferentially binds IGF-II and inhibits its actions. The aim of this study was to investigate whether O glycosylation modulates the pharmacokinetics of IGFBP-6. DESIGN AND METHODS: The pharmacokinetic profiles of (125)I-labelled glycosylated (g) and non-glycosylated (n-g) recombinant human IGFBP-6 were studied following intravenous bolus administration in anaesthetised rats. RESULTS: The redistribution half-life of gIGFBP-6 was 2.3-fold greater than that of n-gIGFBP-6 (14.4+/- 1.2 vs 6.3+/-1.5 min, P=0. 006). The elimination half-life of gIGFBP-6 was 21-fold greater than that of n-gIGFBP-6 (584.2+/-130.2 vs 28.0+/-4.2 min, P=0.019). The effect of O glycosylation on IGFBP-6 pharmacokinetics was not due to inhibition of intravascular proteolysis. Radioactivity was found in stomach, kidneys, lung, spleen, heart and liver but not brain 4h after injection of g or n-gIGFBP-6. CONCLUSIONS: O-glycosylation delays the clearance of IGFBP-6 from the circulation and may therefore contribute to its role as a circulating inhibitor of IGF-II actions. PMID- 10802532 TI - Recombinant human growth hormone enhances tibial growth in peripubertal female rats but not in males. AB - OBJECTIVE: A novel non-invasive technique termed microknemometry, which allows daily leg length measurement, was used to investigate the growth promoting effect of growth hormone (GH) on peripubertal rats. We compared the effect of different patterns of recombinant human (rh) GH administration to peripubertal male rats with the effect produced by two daily administrations of the same amount of rhGH to peripubertal female rats or adult male rats. Another group of peripubertal male rats was also submitted to a 3-day period of starvation, in order to study catch-up growth during refeeding and to determine whether this process could be stimulated by exogenous GH administration. RESULTS: GH treatment was unable to stimulate tibial growth or weight gain in peripubertal males, whereas a clear growth promoting effect was observed in female rats and also in adult male rats. Starvation caused a dramatic body weight loss, and a reduction in tibial growth rate. Peripubertal male rats gained body weight faster than unstarved animals during refeeding, although recovery was not complete after nine days. Tibial growth, however, was resumed at the same speed as in normally fed males. This means that no catch-up effect was observed after refeeding in animals either with or without GH treatment. CONCLUSIONS: During peripuberty, normal male rats grow at a maximal speed that cannot be further increased by exogenous GH treatment, whereas age-matched female rats or older males grow at a slower rate than peripubertal males. Thus, exogenous rhGH administration is capable of enhancing growth velocity. PMID- 10802534 TI - Three mRNA transcripts of the proopiomelanocortin gene in human placenta at term. AB - The proopiomelanocortin (POMC) gene whose normal pituitary specific mRNA product is 1200 bases (b) is also expressed in placenta and its peptide derivatives such as ACTH and beta-endorphin may play an important role in the initiation of labor. So far, two mRNA transcripts, one small (800b) and one large (1380b) have been reported in placenta by Northern blot analysis, similar to other endocrine tissues and various extrapituitary tumors; however, it is questionable whether both of these transcripts are effectively translated to a functional protein. We examined by Northern blot analysis the size and the differential expression of placental POMC gene transcripts in pregnant women with different modes of delivery. Placental tissues were collected from two groups of pregnant women, six with vaginal delivery (VD) and five with cesarean section (CS). In both groups of placentae three POMC gene transcripts were detected of 800, 1200 and 1380 bases; the 1200b pituitary specific species often predominated and was always present. The 800b transcript was also always present, while the large transcript (1380b) was expressed in 3/6 VD and 2/5 CS placental tissues. No differences in the relative levels of any of these mRNA species showing effect of the mode of delivery were observed. We conclude that POMC gene transcription in placental tissue at term gives rise to three mRNA transcripts, thus resembling extrapituitary tumors. The reported changes in the levels of the derivative peptides according to the mode of delivery do not reflect changes in POMC mRNA levels and could be attributed to a post-translational effect. PMID- 10802533 TI - Atrial natriuretic peptide and aldosterone secretions, and atrial natriuretic peptide-binding sites in kidneys and adrenal glands of pregnant and fetal rats in late gestation in response to a high-salt diet. AB - OBJECTIVE: This study aimed at determining, in the term pregnant rat, whether maternal and fetal plasma atrial natriuretic peptide (ANP) concentrations were modified in response to an oral sodium load, and to investigate whether any changes in plasma concentrations were able to modify the density and affinity of the different ANP-binding site subtypes in maternal and fetal kidneys and adrenal glands. METHODS: Pregnant rats kept in metabolic cages were divided into two groups. The normal sodium diet group had free access to rat chow and tap water whereas the high sodium diet group received 1% NaCl as drinking water for 10 consecutive days from day 11 to day 21 of gestation with free access to standard rat chow. Pregnant rats from both groups were killed by decapitation on day 21 of gestation. The plasma ANP and aldosterone concentrations were determined by RIA. The density and affinity of ANP receptors were determined in the maternal and fetal adrenal glands and kidneys. RESULTS: In the pregnant rats on the high-salt diet, the sodium and water intakes, as well as the urine volume and sodium excretion, were significantly higher than in the control group. After 10 days of high-salt intake, water and sodium retentions were not significantly different in the two groups, indicating that the pregnant rats were able to excrete excess salt. The high sodium intake did not change the body weight of the pregnant rats but did increase the body weight of the fetal rats. Maternal and fetal hematocrits remained unchanged in both groups, the high sodium intake did not modify plasma sodium concentration in the maternal rats but increased that of the fetuses, indicating an accumulation of sodium in the fetal rats. The dietary sodium intake did not change the plasma ANP concentrations but significantly decreased the plasma aldosterone concentrations in both the maternal and fetal rats. In response to the high-salt diet, the density and affinity of total ANP, ANPb and ANPc receptors were not altered in the maternal isolated renal glomeruli or the adrenal zona glomerulosa membranes or the fetal adrenal gland and kidney membrane preparations. CONCLUSION: These results suggest that ANP is not involved in the regulation of water and electrolyte balance in maternal and fetal rats during salt-loaded intake. PMID- 10802535 TI - Comparative analysis of follistatin-, activin beta A- and activin beta B-mRNA steady-state levels in diverse porcine tissues by multiplex S1 nuclease analysis. AB - OBJECTIVE: The relation of activins (dimers of the beta-subunits of inhibin) and follistatin (FS) (their binding protein) affect the growth and differentiation of many cell types. Activin- and FS-mRNAs show a widespread co-expression throughout the organism, indicating an essential role for the FS/activin system in diverse physiological processes. The present study was performed to investigate FS-, activin betaA-, and activin beta B-mRNA expression in porcine tissues and to compare the relative mRNA tissue distribution by a newly developed multiplex S1 nuclease protection assay. METHODS: Twenty micrograms total RNA from different porcine tissues were subjected to multiplex S1 analysis. Specific mRNA expression was determined by measurements of optical densities on autoradiographs. RESULTS: Activin beta A-mRNA expression was abundant in the ovary, adrenal gland, fat, vein, artery and uterus, activin beta B-mRNA was highly expressed in the ovary, pituitary, uterus, placenta, aorta and cerebellum. FS-mRNA showed a widespread expression with high levels in ovary, uterus, cerebellum, placenta and fat. The comparison of relative activin beta A-, activin beta B- and FS-mRNA expression within a certain tissue showed a predominance of activin beta A-mRNA in the adrenal gland, fat, artery, spinal cord, cerebrum and colon and of activin beta B m RNA in pituitary, testis and placenta, while FS-mRNA levels exceeded those of activin subunits in epididymis, liver, lymphoid tissue, muscle, intestine, cerebellum, ovary and uterus. CONCLUSIONS: The presented data provide an overview of FS-, activin beta A-, and activin beta B-mRNA steady state levels in porcine tissues. PMID- 10802536 TI - Crossing the finish line of development: regulated secretion of Dictyostelium proteins. AB - The genesis of the spore coat of Dictyostelium represents an exquisite example of developmentally regulated protein secretion. The proteins that are destined to be assembled into the extracellular matrix of the spore coat are stored in unique prespore vesicles that are triggered to secrete their contents at terminal differentiation. The regulation of this process is being revealed by the identification of the individual proteins in these vesicles. PMID- 10802537 TI - Staufen: a common component of mRNA transport in oocytes and neurons? AB - Mammalian homologues of Staufen, a protein involved in localizing mRNAs during oogenesis and early central nervous system development in Drosophila, have been identified recently. The mammalian staufen gene encodes a protein containing several conserved double-stranded mRNA-binding domains and is expressed in hippocampal neurons. The mammalian Staufen protein forms granules that are transported to the distal dendrite during neuronal maturation. The Staufen granules colocalize with ribonuclear particles that transport mRNA to the dendrites. These findings might provide clues to a mechanism of mRNA transport conserved in mammalian neurons and Drosophila oogenesis. PMID- 10802538 TI - Towards a molecular understanding of cytokinesis. AB - In this review, we focus on recent discoveries regarding the molecular basis of cleavage furrow positioning and contractile ring assembly and contraction during cytokinesis. However, some of these mechanisms might have different degrees of importance in different organisms. This synthesis attempts to uncover common themes and to reveal potential relationships that might contribute to the biochemical and mechanical aspects of cytokinesis. Because the information about cytokinesis is still fairly rudimentary, our goal is not to present a definitive model but to present testable hypotheses that might lead to a better mechanistic understanding of the process. PMID- 10802539 TI - In and out of the postsynaptic region: signalling proteins on the move. AB - Reversible translocation of signalling proteins to and from their sites of action has emerged as an important theme in signal transduction. The recent findings of the stimulus-induced translocation of Ca2+-calmodulin-dependent protein kinase II (CaMKII) and the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor to and from the postsynaptic region are model cases for understanding how the dynamic localization of signalling proteins is used to regulate signal transduction. PMID- 10802540 TI - Triggering the cell cycle in plants. AB - In essence, the mitotic cell cycle in eukaryotes involves the duplication and separation of chromosomes, coupled to the process of dividing one cell into two. Cytokinesis is therefore the culmination of a series of events that were triggered during G1 phase, and brings the daughter cells back to the starting position in G1 for another possible round of division. In all eukaryotes, progression through the cell cycle is controlled by cyclin-dependent kinases that bind to positive regulators called cyclins. This review explores some of the pathways that trigger the plant cell cycle, with emphasis on the G1 phase. Examples include signalling pathways involving glutathione and cellular redox potential, the possible existence of a G1 DNA-damage checkpoint, and the plant hormones auxin and cytokinin. Progress in understanding the link between cell proliferation, cell differentiation and the cell-cycle machinery in a developmental context is discussed. PMID- 10802541 TI - Protein complexes in transport vesicle targeting. AB - The transport of material between membrane-bounded organelles in eukaryotic cells requires the accurate delivery of different classes of carrier vesicles to specific target compartments. Recent studies indicate that different targeting reactions involve distinct protein complexes that act to mark the target organelle for incoming vesicles. This review focuses on the proteins and protein complexes that have been implicated in various targeting reactions. PMID- 10802542 TI - Kai Simons- a new millennium, a new direction. PMID- 10802543 TI - Careers-perspective interview. PMID- 10802545 TI - Snakes: a new 'model organism' in ecological research? PMID- 10802544 TI - Shape distances, shape spaces and the comparison of morphometric methods. PMID- 10802546 TI - Biodiversity in groundwater: a large-scale view. PMID- 10802547 TI - Experimental tests of sex-allocation theory in plants. AB - A general explanation for diversity in plant breeding systems is offered by sex allocation theory. This theory assumes a trade-off between allocation of resources to the two sexual functions. It explains the high frequency of hermaphroditism in angiosperms by diminishing fitness returns on investment of more resources in a single function. Recent experimental studies provide tests of this theory by measuring male and female fitness gains, and examining the trade off assumption. These studies show how fitness responds to shifts in allocation. Allocation traits often show heritable variation, but support for a trade-off remains weak. PMID- 10802548 TI - Pulsed resources and community dynamics of consumers in terrestrial ecosystems. AB - Many terrestrial ecosystems are characterized by intermittent production of abundant resources for consumers, such as mast seeding and pulses of primary production following unusually heavy rains. Recent research is revealing patterns in the ways that consumer communities respond to these pulsed resources. Studies of the ramifying effects of pulsed resources on consumer communities integrate 'top-down' and 'bottom-up' approaches to community dynamics, and illustrate how the strength of species interactions can change dramatically through time. PMID- 10802549 TI - The role of polyphenols in terrestrial ecosystem nutrient cycling. AB - Interspecific variation in polyphenol production by plants has been interpreted in terms of defense against herbivores. Several recent lines of evidence suggest that polyphenols also influence the pools and fluxes of inorganic and organic soil nutrients. Such effects could have far-ranging consequences for nutrient competition among and between plants and microbes, and for ecosystem nutrient cycling and retention. The significance of polyphenols for nutrient cycling and plant productivity is still uncertain, but it could provide an alternative or complementary explanation for the variability in polyphenol production by plants. PMID- 10802550 TI - The emergence and maintenance of diversity: insights from experimental bacterial populations. AB - Mechanisms maintaining genetic and phenotypic variation in natural populations are central issues in ecology and evolution. However, the long generation times of most organisms and the complexity of natural environments have made elucidation of ecological and evolutionary mechanisms difficult. Experiments using bacterial populations propagated in controlled environments reduce ecosystem complexity to the point where understanding simple processes in isolation becomes possible. Recent studies reveal the circumstances and mechanisms that promote the emergence of stable polymorphisms. PMID- 10802551 TI - Are all mass invasions alike? PMID- 10802552 TI - Are all mass invasions alike? PMID- 10802553 TI - Reply from A. Ricciardi and H.J. MacIsaac. PMID- 10802554 TI - A constrained view of constraints. PMID- 10802555 TI - Reply from M. Pigliucci and J. Kaplan. PMID- 10802556 TI - Bimodal hybrid zones and speciation. AB - Contact zones exemplify a series of stages in speciation. In unimodal hybrid zones intermediates predominate; in bimodal zones hybrids are rare and parental forms predominate; and finally, species might overlap, but never hybridize. Recent studies show bimodality to be associated strongly with assortative mating or fertilization, and only weakly with overall levels of genetic divergence or intrinsic genomic incompatibility. Ecological divergence across most bimodal hybrid zones suggests that ecology contributes more to speciation than genomic incompatibility. This continuum of stable contact zones provides empirical evidence for a route to speciation, which does not require allopatry. PMID- 10802557 TI - Biotechnology food standards hunger for scientific rationale. PMID- 10802558 TI - Less is better: new approaches for seedless fruit production. PMID- 10802559 TI - Chemical synthesis of proteins. AB - The manipulation of protein structure enables a better understanding of the principles of protein folding, as well as the development of novel therapeutics and drug-delivery vehicles. Chemical synthesis is the most powerful approach for constructing proteins of novel design and structure, allowing for variation of covalent structure without limitations. Here we describe the various chemical methods that are currently used for creating proteins of unique architecture and function. PMID- 10802560 TI - The synthesis of chiral cyanohydrins by oxynitrilases. AB - Enantiomerically pure cyanohydrins are important synthetic intermediates for pharmaceuticals and agrochemicals. They are produced by enzyme-catalysed synthesis using oxynitrilases. Sufficient quantities of enzyme are available via cheap natural sources and there have been recent advances in overexpression production of cyanohydrins on an industrial scale. PMID- 10802561 TI - Engineering chloroplasts: an alternative site for foreign genes, proteins, reactions and products. AB - Plant genetic engineering via the nucleus is a mature technology that has been used very productively for research and commercial biotechnology. By contrast, the ability to introduce foreign genes at specific locations on a chloroplast's chromosome has been acquired relatively recently. Certain limitations of nuclear genome transformation methods might be overcome by the site-specific introduction of genes into plastid chromosomes. In addition, plastids, mitochondria and other subcellular organelles might provide more favorable environments than the nuclear cytoplasmic compartment for certain biochemical reactions and for accumulating large amounts of some gene and enzyme products. PMID- 10802562 TI - Regulation and manipulation of the gene clusters encoding type-I PKSs. AB - Modular polyketide synthases are large, multifunctional enzyme complexes that are involved in the biosynthesis of important polyketides. Recent studies have revolutionized our understanding of the linear organization of polyketide synthase-gene clusters. They have provided crucial information on the initiation, elongation and termination of polyketide chains, and thus a rational basis for the generation of novel compounds. Combinatorial libraries have helped this field to move from a random approach to a more empirical phase. The large number of diverse analogs of antibiotics that are presently produced demonstrate the enormous potential of combinatorial biosynthesis. PMID- 10802563 TI - Accidental death of a bubble. PMID- 10802564 TI - Intellectual propriety. PMID- 10802566 TI - Reply to "Anhydrobiotic engineering" PMID- 10802565 TI - The practical problems of post-genomic biology. PMID- 10802569 TI - Therapeutic cloning OK PMID- 10802567 TI - Anhydrobiotic engineering. PMID- 10802570 TI - GM roundup PMID- 10802571 TI - Two SNP deals for gemini PMID- 10802572 TI - Genome debate continues PMID- 10802573 TI - Anti-biotech protest PMID- 10802574 TI - DuPont sues monsanto PMID- 10802576 TI - USDA testing lab planned PMID- 10802575 TI - Initial public offerings, 1st quarter 2000 PMID- 10802578 TI - Biopharmaceutical sector performance, 1st quarter 2000 PMID- 10802579 TI - The advisory committee strikes back PMID- 10802577 TI - Public GM plant safety program PMID- 10802581 TI - Biotech info campaign PMID- 10802580 TI - Cartoon PMID- 10802582 TI - Gene chip battle PMID- 10802583 TI - Biotech fundraising, Q1 2000 PMID- 10802584 TI - Research collaborations PMID- 10802585 TI - Transgenic pest control on the fly PMID- 10802586 TI - Genetic testing of the other half PMID- 10802587 TI - New antibiotics show some backbone PMID- 10802588 TI - Array for membrane proteins PMID- 10802590 TI - Drawing a bead on bioassays PMID- 10802589 TI - Drug smugglers come under umbrella PMID- 10802591 TI - AAV divides and conquers PMID- 10802592 TI - Screening protein libraries for new activities PMID- 10802593 TI - Feedback regulation in metabolic engineering PMID- 10802595 TI - Immunostimulatory DNA vaccines give tumor therapy a boost PMID- 10802596 TI - Antisense branches into high amylose starch PMID- 10802594 TI - Antisense treatment for psoriasis PMID- 10802598 TI - Chimeric oligonucleotides-a tool for crop improvement? PMID- 10802597 TI - A tool for identifying proteasome inhibitors PMID- 10802600 TI - Technical report PMID- 10802599 TI - Review PMID- 10802601 TI - Tapping into tumor immunotherapy PMID- 10802602 TI - First round to Amgen in EPO battle. PMID- 10802603 TI - GM food regulatory terms debated. PMID- 10802605 TI - NAS report: strengthen agbio regs and relations. National Academy of Sciences. PMID- 10802604 TI - Monsanto releases rice data to academia. PMID- 10802606 TI - USDA biotech advisory panel plots uncertain course PMID- 10802607 TI - Biotech companies adapt to big pharma mergers. PMID- 10802608 TI - EU Court overrules France's Bt maize ban. PMID- 10802609 TI - Neuer Markt drives IPOs in Europe. PMID- 10802610 TI - Experts disagree over color of biomass. PMID- 10802611 TI - Problem solving for tumor immunotherapy. PMID- 10802612 TI - In defense of complete genomes. PMID- 10802614 TI - Expanding the AAV package. PMID- 10802613 TI - Probing the proteasome pathway. PMID- 10802615 TI - Public biotech 99: the numbers. PMID- 10802616 TI - The use of single-nucleotide polymorphism maps in pharmacogenomics. AB - Single-nucleotide polymorphisms (SNPs), common variations among the DNA of individuals, are being uncovered and assembled into large SNP databases that promise to enable the dissection of the genetic basis of disease and drug response (i.e., pharmacogenomics). Although great strides have been made in understanding the diversity of the human genome, such as the frequency, distribution, and type of genetic variation that exists, the feasibility of applying this information to uncover useful pharmacogenomic markers is uncertain. The health care industry is clamoring for access to SNP databases for use in research in the hope of revolutionizing the drug development process. As the reality of using SNPs to uncover drug response markers is rarely addressed, this review discusses practical issues, such as patient sample size, SNP density and genome coverage, and data interpretation, that will be important for determining the applicability of pharmacogenomic information to medical practice. PMID- 10802617 TI - Immunostimulatory DNA-based vaccines induce cytotoxic lymphocyte activity by a T helper cell-independent mechanism. AB - Immunostimulatory DNA sequences (ISS) contain unmethylated CpG dinucleotides within a defined motif. Immunization with ISS-based vaccines has been shown to induce high antigen-specific cytotoxic lymphocyte (CTL) activity and a Th1-biased immune response. We have developed a novel ISS-based vaccine composed of ovalbumin (OVA) chemically conjugated to ISS-oligodeoxynucleotide (ODN). Protein ISS conjugate (PIC) is more potent in priming CTL activity and Th1-biased immunity than other ISS-based vaccines. Cytotoxic lymphocyte activation by ISS ODN-based vaccines is preserved in both CD4-/- and MHC class II-/- gene-deficient animals. Furthermore, PIC provides protection against a lethal burden of OVA expressing tumor cells in a CD8+ cell-dependent manner. These results demonstrate that PIC acts through two unique mechanisms: T-helper-independent activation of CTL and facilitation of exogenous antigen presentation on MHC class I. This technology may have clinical applications in cancer therapy and in stimulating host defense in AIDS and chronic immunosuppression. PMID- 10802618 TI - TAP expression provides a general method for improving the recognition of malignant cells in vivo. AB - A major class of tumors lack expression of the transporters associated with antigen processing (TAP). These proteins are essential for delivery of antigenic peptides into the lumen of the endoplasmic reticulum (ER) and subsequent assembly with nascent major histocompatibility complex (MHC) class I, which results in cell surface presentation of the trimeric complex to cytolytic T lymphocytes. Cytolytic T lymphocytes are major effector cells in immunosurveillance against tumors. Here we have tested the hypothesis that TAP downregulation in tumors allows immunosubversion of this effector mechanism, by establishing a model system to examine the role of TAP in vivo in restoring antigen presentation, immune recognition, and effects on malignancy of the TAP-deficient small-cell lung carcinoma, CMT.64. To test the potential of providing exogenous TAP in cancer therapies, we constructed a vaccinia virus (VV) containing the TAP1 gene and examined whether VV-TAP1 could reduce tumors in mice. The results demonstrate that TAP should be considered for inclusion in cancer therapies, as it is likely to provide a general method for increasing immune responses against tumors regardless of the antigenic complement of the tumor or the MHC haplotypes of the host. PMID- 10802619 TI - Reversal of epidermal hyperproliferation in psoriasis by insulin-like growth factor I receptor antisense oligonucleotides. AB - Epidermal hyperplasia is a key feature of the common skin disorder psoriasis. Stimulation of epidermal keratinocytes by insulin-like growth factor I (IGF-I) is essential for cell division, and increased sensitivity to IGF-I may occur in psoriasis. We hypothesized that inhibition of IGF-I receptor expression in the psoriasis lesion would reverse psoriatic epidermal hyperplasia by slowing the rate of keratinocyte cell division. Here we report the use of C5-propynyl-dU,dC phosphorothioate antisense oligonucleotides to inhibit IGF-I receptor expression in keratinocytes. We identified several inhibitory antisense oligonucleotides and demonstrated IGF-I receptor inhibition in vitro through an mRNA targeting mechanism. Repeated injection of these oligonucleotides into human psoriasis lesions, grafted onto nude mice, caused a dramatic normalization of the hyperplastic epidermis. The findings indicate that IGF-I receptor stimulation is a rate-limiting step in psoriatic epidermal hyperplasia and that IGF-I receptor targeting by cutaneous administration of antisense oligonucleotides forms the basis of a potential new psoriasis therapy. PMID- 10802620 TI - Increasing the size of rAAV-mediated expression cassettes in vivo by intermolecular joining of two complementary vectors. AB - A major shortcoming to the use of adeno-associated virus (rAAV) vectors is their limited packaging size. To overcome this hurdle, we split an expression cassette and cloned it into two separate vectors. The vectors contained either a nuclear localizing Escherichia coli lacZ transgene (nlslacZ) with a splice acceptor, or the human elongation factor 1alpha ( EF1alpha) gene enhancer/promoter(s) (EF1alphaEP) with a splice donor. We co-injected a promoter-less nlslacZ vector with a vector containing either a single EF1alphaEP or a double copy of the EF1alphaEP in a head-to-head orientation, into the portal vein of mice. Gene expression, measured by both transduction efficiency and quantitation of the recombinant protein, was as much as 60-70% of that obtained from mice that received a single vector containing a complete EFalphaEP/nlslacZ expression cassette. This two-vector approach may allow development of gene therapy strategies that will carry exogenous DNA sequences with large therapeutic cDNAs and/or regulatory elements. PMID- 10802621 TI - Improving lycopene production in Escherichia coli by engineering metabolic control. AB - Metabolic engineering has achieved encouraging success in producing foreign metabolites in a variety of hosts. However, common strategies for engineering metabolic pathways focus on amplifying the desired enzymes and deregulating cellular controls. As a result, uncontrolled or deregulated metabolic pathways lead to metabolic imbalance and suboptimal productivity. Here we have demonstrated the second stage of metabolic engineering effort by designing and engineering a regulatory circuit to control gene expression in response to intracellular metabolic states. Specifically, we recruited and altered one of the global regulatory systems in Escherichia coli, the Ntr regulon, to control the engineered lycopene biosynthesis pathway. The artificially engineered regulon, stimulated by excess glycolytic flux through sensing of an intracellular metabolite, acetyl phosphate, controls the expression of two key enzymes in lycopene synthesis in response to flux dynamics. This intracellular control loop significantly enhanced lycopene production while reducing the negative impact caused by metabolic imbalance. Although we demonstrated this strategy for metabolite production, it can be extended into other fields where gene expression must be closely controlled by intracellular physiology, such as gene therapy. PMID- 10802622 TI - Short-lived green fluorescent proteins for quantifying ubiquitin/proteasome dependent proteolysis in living cells. AB - The ubiquitin/proteasome-dependent proteolytic pathway is an attractive target for therapeutics because of its critical involvement in cell cycle progression and antigen presentation. However, dissection of the pathway and development of modulators are hampered by the complexity of the system and the lack of easily detectable authentic substrates. We have developed a convenient reporter system by producing N-end rule and ubiquitin fusion degradation (UFD)-targeted green fluorescent proteins that allow quantification of ubiquitin/proteasome-dependent proteolysis in living cells. Accumulation of these reporters serves as an early predictor of G2/M arrest and apoptosis in cells treated with proteasome inhibitors. Comparison of reporter accumulation and cleavage of fluorogenic substrates demonstrates that the rate-limiting chymotrypsin-like activity of the proteasome can be substantially curtailed without significant effect on ubiquitin dependent proteolysis. These reporters provide a new powerful tool for elucidation of the ubiquitin/proteasome pathway and for high throughput screening of compounds that selectively modify proteolysis in vivo. PMID- 10802623 TI - Using an AraC-based three-hybrid system to detect biocatalysts in vivo. AB - Recent methods to create large libraries of proteins have greatly advanced the discovery of proteins with novel functions. However, one limitation in the discovery of new biocatalysts is the screening or selection methods employed to find enzymes from these libraries. We have developed a potentially general method termed QUEST (QUerying for EnzymeS using the Three-hybrid system), which allows the construction of an easily screened or selected phenotype for, in theory, any type of enzymatic reaction. The method couples the in vivo concentration of an enzyme's substrate to changes in the transcriptional level of a reporter operon. Using the arabinose operon activator AraC, we constructed a system capable of detecting the fungal enzyme scytalone dehydratase (SD) in bacteria, and demonstrated its sensitivity and usefulness in library screening. PMID- 10802624 TI - A new microvolume technique for bioaffinity assays using two-photon excitation. AB - Bioaffinity binding assays such as the immunoassay are widely used in life science research. In an immunoassay, specific antibodies are used to bind target molecules in the sample, and quantification of the binding reaction reveals the amount of the target molecules. Here we present a method to measure bioaffinity assays using the two-photon excitation of fluorescence. In this method, microparticles are used as solid phase in binding the target molecules. The degree of binding is then quantified from individual microparticles by use of two photon excitation of fluorescence. We demonstrated the effectiveness of the method using the human alpha-fetoprotein (AFP) immunoassay, which is used to detect fetal disorders. The sensitivity and dynamic range we obtained with this assay indicate that this method can provide a cost-effective and simple way to measure various biomolecules in solution for research and clinical applications. PMID- 10802625 TI - Production of very-high-amylose potato starch by inhibition of SBE A and B. AB - High-amylose starch is in great demand by the starch industry for its unique functional properties. However, very few high-amylose crop varieties are commercially available. In this paper we describe the generation of very-high amylose potato starch by genetic modification. We achieved this by simultaneously inhibiting two isoforms of starch branching enzyme to below 1% of the wild-type activities. Starch granule morphology and composition were noticeably altered. Normal, high-molecular-weight amylopectin was absent, whereas the amylose content was increased to levels comparable to the highest commercially available maize starches. In addition, the phosphorus content of the starch was increased more than fivefold. This unique starch, with its high amylose, low amylopectin, and high phosphorus levels, offers novel properties for food and industrial applications. PMID- 10802626 TI - Engineering herbicide-resistant maize using chimeric RNA/DNA oligonucleotides. AB - Maize plants resistant to imidazolinone herbicides were engineered through targeted modification of endogenous genes using chimeric RNA/DNA oligonucleotides. A precise single-point mutation was introduced into genes encoding acetohydroxyacid synthase (AHAS), at a position known to confer imidazolinone resistance. Phenotypically normal plants from the converted events (C0) were regenerated from resistant calli and grown to maturity. Herbicide leaf painting confirmed the resistance phenotype in C0 plants and demonstrated the anticipated segregation pattern in C1 progeny. DNA cloning and sequencing of the targeted region in resistant calli and derived C0 and C1 plants confirmed the expected mutation. These results demonstrate that oligonucleotide-mediated gene manipulation can be applied to crop improvement. This approach does not involve genomic integration of transgenes. Since the new trait is obtained through modifying a gene within its normal chromosomal context, position effects, transgene silencing, or other concerns that arise as part of developing transgenic events are avoided. PMID- 10802633 TI - Maximizing profits through intelligent planning and implementation. PMID- 10802632 TI - Universal DNA array detection of small insertions and deletions in BRCA1 and BRCA2. AB - Array-based mutation detection methodology typically relies on direct hybridization of the fluorescently labeled query sequence to surface-bound oligonucleotide probes. These probes contain either small sequence variations or perfect-match sequence. The intensity of fluorescence bound to each oligonucleotide probe is intended to reveal which sequence is perfectly complementary to the query sequence. However, these approaches have not always been successful, especially for detection of small frameshift mutations. Here we describe a multiplex assay to detect small insertions and deletions by using a modified PCR to evenly amplify each amplicon (PCR/PCR), followed by ligase detection reaction (LDR). Mutations were identified by screening reaction products with a universal DNA microarray, which uncouples mutation detection from array hybridization and provides for high sensitivity. Using the three BRCA1 and BRCA2 founder mutations in the Ashkenazi Jewish population (BRCA1 185delAG; BRCA1 5382insC; BRCA2 6174delT) as a model system, the assay readily detected these mutations in multiplexed reactions. Our results demonstrate that universal microarray analysis of PCR/PCR/LDR products permits rapid identification of small insertion and deletion mutations in the context of both clinical diagnosis and population studies. PMID- 10802634 TI - Recent patents in green fluorescent protein research PMID- 10802635 TI - People PMID- 10802636 TI - New products PMID- 10802637 TI - Spin-offs versus start-ups as business models in biotechnology. PMID- 10802638 TI - Sex, genes and women's health. PMID- 10802639 TI - Robustness--it's not where you think it is. PMID- 10802640 TI - Maintaining imprinting. PMID- 10802642 TI - Touchingbase PMID- 10802641 TI - Triglycerides and toggling the tummy. PMID- 10802643 TI - OMIM passes the 1,000-disease-gene mark. PMID- 10802644 TI - Allegro, a new computer program for multipoint linkage analysis. PMID- 10802645 TI - The gene TFR2 is mutated in a new type of haemochromatosis mapping to 7q22. AB - Haemochromatosis is a common recessive disorder characterized by progressive iron overload, which may lead to severe clinical complications. Most patients are homozygous for the C282Y mutation in HFE on 6p (refs 1-5). A locus for juvenile haemochromatosis (HFE2) maps to 1q (ref. 7). Here we report a new locus (HFE3) on 7q22 and show that a homozygous nonsense mutation in the gene encoding transferrin receptor-2 (TFR2) is found in people with haemochromatosis that maps to HFE3. PMID- 10802646 TI - Missense mutations in MIP underlie autosomal dominant 'polymorphic' and lamellar cataracts linked to 12q. AB - Human inherited cataract is both clinically diverse and genetically heterogeneous. Here we report the identification of the first mutations affecting the major intrinsic protein of the lens, MIP, encoded by the gene MIP on 12q14. MIP is a member of the aquaporin family of membrane-bound water channels. The mutations identified are predicted to disturb water flux across the lens cell membrane. PMID- 10802647 TI - Charcot-Marie-Tooth type 4B is caused by mutations in the gene encoding myotubularin-related protein-2. AB - A gene mutated in Charcot-Marie-Tooth disease type 4B (CMT4B), an autosomal recessive demyelinating neuropathy with myelin outfoldings, has been mapped on chromosome 11q22. Using a positional-cloning strategy, we identified in unrelated CMT4B patients mutations occurring in the gene MTMR2, encoding myotubularin related protein-2, a dual specificity phosphatase (DSP). PMID- 10802648 TI - The imprinted antisense RNA at the Igf2r locus overlaps but does not imprint Mas1. AB - The gene encoding the insulin-like growth-factor type-2 receptor (Igf2r) is maternally expressed and imprinted. A CpG island in Igf2r intron 2 that carries a maternal-specific methylation imprint was shown in a transgenic model to be essential for Igf2r imprinting and for the production of an antisense RNA from the paternal allele. We report here that the endogenous region2 is the promoter for this antisense RNA (named Air, for antisense Igf2r RNA) and that the 3' end lies 107,796 bp distant in an intron of the flanking, but non-imprinted, gene Mas1. PMID- 10802649 TI - The helix files PMID- 10802650 TI - New tricks with pond scum PMID- 10802651 TI - Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. PMID- 10802652 TI - Shotgun sample sequence comparisons between mouse and human genomes. AB - A mixed 'clone-by-clone' and 'whole-genome shotgun' strategy will be used to determine the genomic sequence of the mouse. This method will allow a phase of rapid annotation of the contemporaneous human sequence draft, through whole genome 'sample sequence comparisons'. PMID- 10802653 TI - Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system. AB - The development of non-viral gene-transfer technologies that can support stable chromosomal integration and persistent gene expression in vivo is desirable. Here we describe the successful use of transposon technology for the nonhomologous insertion of foreign genes into the genomes of adult mammals using naked DNA. We show that the Sleeping Beauty transposase can efficiently insert transposon DNA into the mouse genome in approximately 5-6% of transfected mouse liver cells. Chromosomal transposition resulted in long-term expression (>5 months) of human blood coagulation factor IX at levels that were therapeutic in a mouse model of haemophilia B. Our results establish DNA-mediated transposition as a new genetic tool for mammals, and provide new strategies to improve existing non-viral and viral vectors for human gene therapy applications. PMID- 10802654 TI - Mutations in TFAP2B cause Char syndrome, a familial form of patent ductus arteriosus. AB - Char syndrome is an autosomal dominant trait characterized by patent ductus arteriosus, facial dysmorphism and hand anomalies. Using a positional candidacy strategy, we mapped TFAP2B, encoding a transcription factor expressed in neural crest cells, to the Char syndrome critical region and identified missense mutations altering conserved residues in two affected families. Mutant TFAP2B proteins dimerized properly in vitro, but showed abnormal binding to TFAP2 target sequence. Dimerization of both mutants with normal TFAP2B adversely affected transactivation, demonstrating a dominant-negative mechanism. Our work shows that TFAP2B has a role in ductal, facial and limb development and suggests that Char syndrome results from derangement of neural-crest-cell derivatives. PMID- 10802655 TI - A common polymorphism acts as an intragenic modifier of mutant p53 behaviour. AB - The p73 protein, a homologue of the tumour-suppressor protein p53, can activate p53-responsive promoters and induce apoptosis in p53-deficient cells. Here we report that some tumour-derived p53 mutants can bind to and inactivate p73. The binding of such mutants is influenced by whether TP53 (encoding p53) codon 72, by virtue of a common polymorphism in the human population, encodes Arg or Pro. The ability of mutant p53 to bind p73, neutralize p73-induced apoptosis and transform cells in cooperation with EJ-Ras was enhanced when codon 72 encoded Arg. We found that the Arg-containing allele was preferentially mutated and retained in squamous cell tumours arising in Arg/Pro germline heterozygotes. Thus, inactivation of p53 family members may contribute to the biological properties of a subset of p53 mutants, and a polymorphic residue within p53 affects mutant behaviour. PMID- 10802656 TI - Combined activation of Ras and Akt in neural progenitors induces glioblastoma formation in mice. AB - Gliomas are the most common primary malignant brain tumours and are classified into four clinical grades, with the most aggressive tumours being grade 4 astrocytomas (also known as glioblastoma multiforme; GBM). Frequent genetic alterations in GBMs (refs 2-5) result in stimulation of common signal transduction pathways involving Ras, Akt and other proteins. It is not known which of these pathways, if any, are sufficient to induce GBM formation. Here we transfer, in a tissue-specific manner, genes encoding activated forms of Ras and Akt to astrocytes and neural progenitors in mice. We found that although neither activated Ras nor Akt alone is sufficient to induce GBM formation, the combination of activated Ras and Akt induces high-grade gliomas with the histological features of human GBMs. These tumours appear to arise after gene transfer to neural progenitors, but not after transfer to differentiated astrocytes. Increased activity of RAS is found in many human GBMs (ref. 11), and we show here that Akt activity is increased in most of these tumours, implying that combined activation of these two pathways accurately models the biology of this disease. PMID- 10802657 TI - Large-scale identification of secreted and membrane-associated gene products using DNA microarrays. AB - Membrane-associated and secreted proteins are an important class of proteins and include receptors, transporters, adhesion molecules, hormones and cytokines. Although algorithms have been developed to recognize potential amino-terminal membrane-targeting signals or transmembrane domains in protein sequences, their accuracy is limited and they require knowledge of the entire coding sequence, including the N terminus, which is not currently available for most of the genes in most organisms, including human. Several experimental approaches for identifying secreted and membrane proteins have been described, but none have taken a comprehensive genomic approach. Furthermore, none of these methods allow easy classification of clones from arrayed cDNA libraries, for which large-scale gene-expression data are now becoming available through the use of DNA microarrays. We describe here a rapid and efficient method for identifying genes that encode secreted or membrane proteins. mRNA species bound to membrane associated polysomes were separated from other mRNAs by sedimentation equilibrium or sedimentation velocity. The distribution of individual transcripts in the 'membrane-bound' and 'cytosolic' fractions was quantitated for thousands of genes by hybridization to DNA microarrays. Transcripts known to encode secreted or membrane proteins were enriched in the membrane-bound fractions, whereas those known to encode cytoplasmic proteins were enriched in the fractions containing mRNAs associated with free and cytoplasmic ribosomes. On this basis, we identified over 275 human genes and 285 yeast genes that are likely to encode previously unrecognized secreted or membrane proteins. PMID- 10802658 TI - Protection of Rpe65-deficient mice identifies rhodopsin as a mediator of light induced retinal degeneration. AB - Light-induced apoptosis of photoreceptors represents an animal model for retinal degeneration. Major human diseases that affect vision, such as age-related macular degeneration (AMD) and some forms of retinitis pigmentosa (RP), may be promoted by light. The receptor mediating light damage, however, has not yet been conclusively identified; candidate molecules include prostaglandin synthase, cytochrome oxidase, rhodopsin, and opsins of the cones and the retinal pigment epithelium (PE). We exposed to bright light two groups of genetically altered mice that lack the visual pigment rhodopsin (Rpe65-/- and Rho-/-). The gene Rpe65 is specifically expressed in the PE and essential for the re-isomerization of all trans retinol in the visual cycle and thus for the regeneration of rhodopsin after bleaching. Rho-/- mice do not express the apoprotein opsin in photoreceptors, which, consequently, do not contain rhodopsin. We show that photoreceptors lacking rhodopsin in these mice are completely protected against light-induced apoptosis. The transcription factor AP-1, a central element in the apoptotic response to light, is not activated in the absence of rhodopsin, indicating that rhodopsin is essential for the generation or transduction of the intracellular death signal induced by light. PMID- 10802659 TI - Rom-1 is required for rod photoreceptor viability and the regulation of disk morphogenesis. AB - The homologous membrane proteins Rom-1 and peripherin-2 are localized to the disk rims of photoreceptor outer segments (OSs), where they associate as tetramers and larger oligomers. Disk rims are thought to be critical for disk morphogenesis, OS renewal and the maintenance of OS structure, but the molecules which regulate these processes are unknown. Although peripherin-2 is known to be required for OS formation (because Prph2-/- mice do not form OSs; ref. 6), and mutations in RDS (the human homologue of Prph2) cause retinal degeneration, the relationship of Rom-1 to these processes is uncertain. Here we show that Rom1-/- mice form OSs in which peripherin-2 homotetramers are localized to the disk rims, indicating that peripherin-2 alone is sufficient for both disk and OS morphogenesis. The disks produced in Rom1-/- mice were large, rod OSs were highly disorganized (a phenotype which largely normalized with age) and rod photoreceptors died slowly by apoptosis. Furthermore, the maximal photoresponse of Rom1-/- rod photoreceptors was lower than that of controls. We conclude that Rom-1 is required for the regulation of disk morphogenesis and the viability of mammalian rod photoreceptors, and that mutations in human ROM1 may cause recessive photoreceptor degeneration. PMID- 10802660 TI - De novo deletions of SNRPN exon 1 in early human and mouse embryos result in a paternal to maternal imprint switch. AB - Prader-Willi syndrome (PWS) is a neurogenetic disease characterized by infantile hypotonia, gonadal hypoplasia, obsessive behaviour and neonatal feeding difficulties followed by hyperphagia, leading to profound obesity. PWS is due to a lack of paternal genetic information at 15q11-q13 (ref. 2). Five imprinted, paternally expressed genes map to the PWS region, MKRN3 (ref. 3), NDN (ref. 4), NDNL1 (ref. 5), SNRPN (refs 6-8 ) and IPW (ref. 9), as well as two poorly characterized framents designated PAR-1 and PAR-5 (ref. 10). Imprinting of this region involves a bipartite 'imprinting centre' (IC), which overlaps SNRPN (refs 10,11). Deletion of the SNRPN promoter/exon 1 region (the PWS IC element) appears to impair the establishment of the paternal imprint in the male germ line and leads to PWS. Here we report a PWS family in which the father is mosaic for an IC deletion on his paternal chromosome. The deletion chromosome has acquired a maternal methylation imprint in his somatic cells. We have made identical findings in chimaeric mice generated from two independent embryonic stem (ES) cell lines harbouring a similar deletion. Our studies demonstrate that the PWS IC element is not only required for the establishment of the paternal imprint, but also for its postzygotic maintenance. PMID- 10802661 TI - Mutation of a gene encoding a putative chaperonin causes McKusick-Kaufman syndrome. AB - McKusick-Kaufman syndrome (MKKS, MIM 236700) is a human developmental anomaly syndrome comprising hydrometrocolpos (HMC), postaxial polydactyly (PAP) and congenital heart disease (CHD). MKKS has been mapped in the Old Order Amish population to 20p12, between D20S162 and D20S894 (ref. 3). Here we describe the identification of a gene mutated in MKKS. We analysed the approximately 450-kb candidate region by sample sequencing, which revealed the presence of several known genes and EST clusters. We evaluated candidate transcripts by northern-blot analysis of adult and fetal tissues. We selected one transcript with widespread expression, MKKS, for analysis in a patient from the Amish pedigree and a sporadic, non-Amish case. The Old Order Amish patient was found to be homozygous for an allele that had two missense substitutions and the non-Amish patient was a compound heterozygote for a frameshift mutation predicting premature protein truncation and a distinct missense mutation. The MKKS predicted protein shows amino acid similarity to the chaperonin family of proteins, suggesting a role for protein processing in limb, cardiac and reproductive system development. We believe that this is the first description of a human disorder caused by mutations affecting a putative chaperonin molecule. PMID- 10802662 TI - Conditional inactivation of Fgf4 reveals complexity of signalling during limb bud development. AB - Development of the vertebrate limb bud depends on reciprocal interactions between the zone of polarizing activity (ZPA) and the apical ectodermal ridge (AER). Sonic hedgehog (SHH) and fibroblast growth factors (FGFs) are key signalling molecules produced in the ZPA and AER, respectively. Experiments in chicks suggested that SHH expression in the ZPA is maintained by FGF4 expression in the AER, and vice versa, providing a molecular mechanism for coordinating the activities of these two signalling centres. This SHH/FGF4 feedback loop model is supported by genetic evidence showing that Fgf4 expression is not maintained in Shh-/- mouse limbs. We report here that Shh expression is maintained and limb formation is normal when Fgf4 is inactivated in mouse limbs, thus contradicting the model. We also found that maintenance of Fgf9 and Fgf17 expression is dependent on Shh, whereas Fgf8 expression is not. We discuss a model in which no individual Fgf expressed in the AER (AER-Fgf) is solely necessary to maintain Shh expression, but, instead, the combined activities of two or more AER-Fgfs function in a positive feedback loop with Shh to control limb development. PMID- 10802663 TI - Obesity resistance and multiple mechanisms of triglyceride synthesis in mice lacking Dgat. AB - Triglycerides (or triacylglycerols) represent the major form of stored energy in eukaryotes. Triglyceride synthesis has been assumed to occur primarily through acyl CoA:diacylglycerol transferase (Dgat), a microsomal enzyme that catalyses the final and only committed step in the glycerol phosphate pathway. Therefore, Dgat has been considered necessary for adipose tissue formation and essential for survival. Here we show that Dgat-deficient (Dgat-/-) mice are viable and can still synthesize triglycerides. Moreover, these mice are lean and resistant to diet-induced obesity. The obesity resistance involves increased energy expenditure and increased activity. Dgat deficiency also alters triglyceride metabolism in other tissues, including the mammary gland, where lactation is defective in Dgat-/- females. Our findings indicate that multiple mechanisms exist for triglyceride synthesis and suggest that the selective inhibition of Dgat-mediated triglyceride synthesis may be useful for treating obesity. PMID- 10802664 TI - Mutations in KERA, encoding keratocan, cause cornea plana. AB - Specialized collagens and small leucine-rich proteoglycans (SLRPs) interact to produce the transparent corneal structure. In cornea plana, the forward convex curvature is flattened, leading to a decrease in refraction. A more severe, recessively inherited form (CNA2; MIM 217300) and a milder, dominantly inherited form (CNA1; MIM 121400) exist. CNA2 is a rare disorder with a worldwide distribution, but a high prevalence in the Finnish population. The gene mutated in CNA2 was assigned by linkage analysis to 12q (refs 4, 5), where there is a cluster of several SLRP genes. We cloned two additional SLRP genes highly expressed in cornea: KERA (encoding keratocan) in 12q and OGN (encoding osteoglycin) in 9q. Here we report mutations in KERA in 47 CNA2 patients: 46 Finnish patients are homozygous for a founder missense mutation, leading to the substitution of a highly conserved amino acid; and one American patient is homozygous for a mutation leading to a premature stop codon that truncates the KERA protein. Our data establish that mutations in KERA cause CNA2. CNA1 patients had no mutations in these proteoglycan genes. PMID- 10802666 TI - Dopamine is required for hyperphagia in Lep(ob/ob) mice. AB - Feeding is a complex process responsive to sensory information related to sight and smell of food, previous feeding experiences, satiety signals elicited by ingestion and hormonal signals related to energy balance. Dopamine released in specific brain regions is associated with pleasurable and rewarding events and may reinforce positive aspects of feeding. Dopamine also influences initiation and coordination of motor activity and is required for sensorimotor functions. Thus, dopamine may facilitate integration of sensory cues related to hunger, initiating the search for food and its consumption. Dopaminergic neurons in the substantia nigra and ventral tegmental area project to the caudate putamen and nucleus accumbens, where they modulate movement and reward. There are projections from the nucleus accumbens to the lateral hypothalamus that regulate feeding. Dopamine-deficient mice (Dbh(Th/+), Th-/-; hereafter DD mice) cannot synthesize dopamine in dopaminergic neurons. They gradually become aphagic and die of starvation. Daily treatment of DD mice with L-3,4-dihydroxyphenylalanine (L-DOPA) transiently restores brain dopamine, locomotion and feeding. Leptin-null (Lep(ob/ob)) mice exhibit obesity, decreased energy expenditure and hyperphagia. As the hypothalamic leptin-melanocortin pathway appears to regulate appetite and metabolism, we generated mice lacking both dopamine and leptin (DD x Lep(ob/ob)) to determine if leptin deficiency overcomes the aphagia of DD mice. DD x Lep(ob/ob) mice became obese when treated daily with L-DOPA, but when L-DOPA treatment was terminated the double mutants were capable of movement, but did not feed. Our data show that dopamine is required for feeding in leptin-null mice. PMID- 10802665 TI - Transcript imaging of the development of human T helper cells using oligonucleotide arrays. AB - Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation. Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes. Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells. PMID- 10802667 TI - Mice deficient in Six5 develop cataracts: implications for myotonic dystrophy. AB - Expansion of a CTG trinucleotide repeat in the 3' UTR of the gene DMPK at the DM1 locus on chromosome 19 causes myotonic dystrophy, a dominantly inherited disease characterized by skeletal muscle dystrophy and myotonia, cataracts and cardiac conduction defects. Targeted deletion of Dm15, the mouse orthologue of human DMPK, produced mice with a mild myopathy and cardiac conduction abnormalities, but without other features of myotonic dystrophy, such as myotonia and cataracts. We, and others, have demonstrated that repeat expansion decreases expression of the adjacent gene SIX5 (refs 7,8), which encodes a homeodomain transcription factor. To determine whether SIX5 deficiency contributes to the myotonic dystrophy phenotype, we disrupted mouse Six5 by replacing the first exon with a beta-galactosidase reporter. Six5-mutant mice showed reporter expression in multiple tissues, including the developing lens. Homozygous mutant mice had no apparent abnormalities of skeletal muscle function, but developed lenticular opacities at a higher rate than controls. Our results suggest that SIX5 deficiency contributes to the cataract phenotype in myotonic dystrophy, and that myotonic dystrophy represents a multigenic disorder. PMID- 10802668 TI - Heterozygous loss of Six5 in mice is sufficient to cause ocular cataracts. AB - Myotonic dystrophy (DM) is an autosomal dominant disorder characterized by skeletal muscle wasting, myotonia, cardiac arrhythmia, hyperinsulinaemia, mental retardation and ocular cataracts. The genetic defect in DM is a CTG repeat expansion located in the 3' untranslated region of DMPK and 5' of a homeodomain encoding gene, SIX5 (formerly DMAHP; refs 2-5). There are three mechanisms by which CTG expansion can result in DM. First, repeat expansion may alter the processing or transport of the mutant DMPK mRNA and consequently reduce DMPK levels. Second, CTG expansion may establish a region of heterochromatin 3' of the repeat sequence and decrease SIX5 transcription. Third, toxic effects of the repeat expansion may be intrinsic to the repeated elements at the level of DNA or RNA (refs 10,11). Previous studies have demonstrated that a dose-dependent loss of Dm15 (the mouse DMPK homologue) in mice produces a partial DM phenotype characterized by decreased development of skeletal muscle force and cardiac conduction disorders. To test the role of Six5 loss in DM, we have analysed a strain of mice in which Six5 was deleted. Our results demonstrate that the rate and severity of cataract formation is inversely related to Six5 dosage and is temporally progressive. Six5+/- and Six5-/- mice show increased steady-state levels of the Na+/K+-ATPase alpha-1 subunit and decreased Dm15 mRNA levels. Thus, altered ion homeostasis within the lens may contribute to cataract formation. As ocular cataracts are a characteristic feature of DM, these results demonstrate that decreased SIX5 transcription is important in the aetiology of DM. Our data support the hypothesis that DM is a contiguous gene syndrome associated with the partial loss of both DMPK and SIX5. PMID- 10802669 TI - ATM-dependent phosphorylation of nibrin in response to radiation exposure. AB - Mutations in the gene ATM are responsible for the genetic disorder ataxia telangiectasia (A-T), which is characterized by cerebellar dysfunction, radiosensitivity, chromosomal instability and cancer predisposition. Both the A-T phenotype and the similarity of the ATM protein to other DNA-damage sensors suggests a role for ATM in biochemical pathways involved in the recognition, signalling and repair of DNA double-strand breaks (DSBs). There are strong parallels between the pattern of radiosensitivity, chromosomal instability and cancer predisposition in A-T patients and that in patients with Nijmegen breakage syndrome (NBS). The protein defective in NBS, nibrin (encoded by NBS1), forms a complex with MRE11 and RAD50 (refs 1,2). This complex localizes to DSBs within 30 minutes after cellular exposure to ionizing radiation (IR) and is observed in brightly staining nuclear foci after a longer period of time. The overlap between clinical and cellular phenotypes in A-T and NBS suggests that ATM and nibrin may function in the same biochemical pathway. Here we demonstrate that nibrin is phosphorylated within one hour of treatment of cells with IR. This response is abrogated in A-T cells that either do not express ATM protein or express near full-length mutant protein. We also show that ATM physically interacts with and phosphorylates nibrin on serine 343 both in vivo and in vitro. Phosphorylation of this site appears to be functionally important because mutated nibrin (S343A) does not completely complement radiosensitivity in NBS cells. ATM phosphorylation of nibrin does not affect nibrin-MRE11-RAD50 association as revealed by radiation induced foci formation. Our data provide a biochemical explanation for the similarity in phenotype between A-T and NBS. PMID- 10802670 TI - Genetic and epigenetic incompatibilities underlie hybrid dysgenesis in Peromyscus. AB - Crosses between the two North American rodent species Peromyscus polionotus (PO) and Peromyscus maniculatus (BW) yield parent-of-origin effects on both embryonic and placental growth. The two species are approximately the same size, but a female BW crossed with a male PO produces offspring that are smaller than either parent. In the reciprocal cross, the offspring are oversized and typically die before birth. Rare survivors are exclusively female, consistent with Haldane's rule, which states that in instances of hybrid sterility or inviability, the heterogametic sex tends to be more severely affected. To understand these sex- and parent-of-origin-specific patterns of overgrowth, we analysed reciprocal backcrosses. Our studies reveal that hybrid inviability is partially due to a maternally expressed X-linked PO locus and an imprinted paternally expressed autosomal BW locus. In addition, the hybrids display skewing of X-chromosome inactivation in favour of the expression of the BW X chromosome. The most severe overgrowth is accompanied by widespread relaxation of imprinting of mostly paternally expressed genes. Both genetic and epigenetic mechanisms underlie hybrid inviability in Peromyscus and hence have a role in the establishment and maintenance of reproductive isolation barriers in mammals. PMID- 10802674 TI - Correction to "NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome" PMID- 10802675 TI - The need for private-public partnerships. PMID- 10802676 TI - Transgenic expression of green fluorescence protein can cause dilated cardiomyopathy. PMID- 10802677 TI - Human neural progenitor cells: better blue than green? PMID- 10802679 TI - Sacked Spanish oncologists win reprieve. PMID- 10802678 TI - Reply to "Human neural progenitor cells: better blue than green?" PMID- 10802680 TI - Second HSC researcher sends anonymous 'Olivieri' note. Hospital for Sick Children. PMID- 10802682 TI - New measures introduced to monitor gene therapy trials. PMID- 10802681 TI - UK speculated to permit stem cell research PMID- 10802684 TI - Japanese research center changes career structure PMID- 10802683 TI - NIH researches response to commonly used medicines. PMID- 10802685 TI - NIH revamps HIV vaccine trials unit. PMID- 10802686 TI - HIV researchers upset by Royal Society discussion of 'River theory'. PMID- 10802687 TI - A new salk-UCSD institute? PMID- 10802688 TI - UK plans to encourage physician scientists. PMID- 10802689 TI - Italy creates neuroscience network. PMID- 10802690 TI - Economists propose new strategy for public health management. PMID- 10802691 TI - Ministers pledge to stop TB PMID- 10802692 TI - Not so Fas: Re-evaluating the mechanisms of immune privilege and tumor escape. PMID- 10802693 TI - Still waiting for the revolution PMID- 10802694 TI - Cancer: the evolutionary legacy PMID- 10802695 TI - The fine cytokine line between graft acceptance and rejection. PMID- 10802696 TI - Caspase 8: the killer you can't live without. PMID- 10802697 TI - Chemotherapeutic drugs--more really is not better. PMID- 10802699 TI - Breaking point PMID- 10802698 TI - Steering anti-cancer drugs away from the TRAIL. PMID- 10802700 TI - Bone cancer gain without the pain. PMID- 10802702 TI - A DNA chip off the aging block. PMID- 10802701 TI - Converting hepatocytes to beta-cells--a new approach for diabetes? PMID- 10802703 TI - For want of a disc, the cell was lost.... PMID- 10802704 TI - The sleep-wake switch: A neuronal alarm clock. PMID- 10802705 TI - Research news PMID- 10802706 TI - Mitochondrial control of cell death. AB - In many instances, permeabilization of mitochondrial membranes is a rate-limiting step of apoptotic or necrotic cell demise. This has important consequences for the pathophysiology of cell death, as well as for its pharmacological control. PMID- 10802707 TI - Osteoprotegerin blocks bone cancer-induced skeletal destruction, skeletal pain and pain-related neurochemical reorganization of the spinal cord. AB - Bone cancer pain is common among cancer patients and can have a devastating effect on their quality of life. A chief problem in designing new therapies for bone cancer pain is that it is unclear what mechanisms drive this distinct pain condition. Here we show that osteoprotegerin, a secreted 'decoy' receptor that inhibits osteoclast activity, also blocks behaviors indicative of pain in mice with bone cancer. A substantial part of the actions of osteoprotegerin seems to result from inhibition of tumor-induced bone destruction that in turn inhibits the neurochemical changes in the spinal cord that are thought to be involved in the generation and maintenance of cancer pain. These results demonstrate that excessive tumor-induced bone destruction is involved in the generation of bone cancer pain and that osteoprotegerin may provide an effective treatment for this common human condition. PMID- 10802708 TI - Caspase 8 is deleted or silenced preferentially in childhood neuroblastomas with amplification of MYCN. AB - Caspase 8 is a cysteine protease regulated in both a death-receptor-dependent and -independent manner during apoptosis. Here, we report that the gene for caspase 8 is frequently inactivated in neuroblastoma, a childhood tumor of the peripheral nervous system. The gene is silenced through DNA methylation as well as through gene deletion. Complete inactivation of CASP8 occurred almost exclusively in neuroblastomas with amplification of the oncogene MYCN. Caspase 8-null neuroblastoma cells were resistant to death receptor- and doxorubicin-mediated apoptosis, deficits that were corrected by programmed expression of the enzyme. Thus, caspase 8 acts as a tumor suppressor in neuroblastomas with amplification of MYCN. PMID- 10802709 TI - Concurrent enteric helminth infection modulates inflammation and gastric immune responses and reduces helicobacter-induced gastric atrophy. AB - Helicobacter pylori is causally associated with gastritis and gastric cancer. Some developing countries with a high prevalence of infection have high gastric cancer rates, whereas in others, these rates are low. The progression of helicobacter-induced gastritis and gastric atrophy mediated by type 1 T-helper cells may be modulated by concurrent parasitic infection. Here, in mice with concurrent helminth infection, helicobacter-associated gastric atrophy was reduced considerably despite chronic inflammation and high helicobacter colonization. This correlated with a substantial reduction in mRNA for cytokines and chemokines associated with a gastric inflammatory response of type 1 T-helper cells. Thus, concurrent enteric helminth infection can attenuate gastric atrophy, a premalignant lesion. PMID- 10802710 TI - Active movement of T cells away from a chemokine. AB - Movement towards or away from a given stimulus guides the directional migration of prokaryotes, simple eukaryotes and neurons. As bi-directional cues may influence entry and exit of immune effector cells from tissue sites, we evaluated the migratory responses of T-cell subsets to varying concentrations of the chemokine stromal cell derived factor-1 (SDF-1). There was selective repulsion of subpopulations of T cells at high concentrations of recombinant SDF-1 or naturally occurring bone marrow-derived SDF-1, which could be inhibited by pertussis toxin and antibody against the chemokine receptor CXCR4. Distinct sensitivity profiles to genistein, herbimycin and 8-Br-cAMP biochemically distinguished movement of cells towards or away from an SDF-1 gradient. In vivo, antigen-induced T-cell recruitment into the peritoneal cavity was reversed by high but not low concentrations of SDF-1. The phenomenon of movement away from a chemokine represents a previously unknown mechanism regulating the localization of mature T cells. It adds to the functional repertoire of chemokines that may participate in immune physiology and may be applied therapeutically to alter the immune response. PMID- 10802711 TI - The axis of interleukin 12 and gamma interferon regulates acute vascular xenogeneic rejection. AB - Recent advances using transgenic animals or exogenous complement inhibitors have demonstrated prevention of hyperacute rejection of vascularized organs, but not graft loss due to acute vascular rejection. Using various wild-type and cytokine deficient mice strains, we have examined the mechanisms of acute vascular rejection. C57BL/6 mice deficient in interleukin12 or gamma interferon showed faster acute vascular rejection than did wild-type mice. Furthermore, mice defective in B-cell development showed no acute vascular rejection. These results demonstrate that the axis of interleukin 12 and gamma interferon provides a survival advantage in vascularized xenografts by delaying or preventing acute vascular rejection caused by a B cell-dependent mechanism. PMID- 10802712 TI - TAK1 is activated in the myocardium after pressure overload and is sufficient to provoke heart failure in transgenic mice. AB - The transforming-growth-factor-beta-activated kinase TAK1 is a member of the mitogen-activated protein kinase kinase kinase family, which couples extracellular stimuli to gene transcription. The in vivo function of TAK1 is not understood. Here, we investigated the potential involvement of TAK1 in cardiac hypertrophy. In adult mouse myocardium, TAK1 kinase activity was upregulated 7 days after aortic banding, a mechanical load that induces hypertrophy and expression of transforming growth factor beta. An activating mutation of TAK1 expressed in myocardium of transgenic mice was sufficient to produce p38 mitogen activated protein kinase phosphorylation in vivo, cardiac hypertrophy, interstitial fibrosis, severe myocardial dysfunction, 'fetal' gene induction, apoptosis and early lethality. Thus, TAK1 activity is induced as a delayed response to mechanical stress, and can suffice to elicit myocardial hypertrophy and fulminant heart failure. PMID- 10802713 TI - Apoptosis induced in normal human hepatocytes by tumor necrosis factor-related apoptosis-inducing ligand. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been reported to induce apoptosis in various tumor cells but not in nontransformed, normal cells. Preclinical studies in mice and nonhuman primates have shown that administration of TRAIL can induce apoptosis in human tumors, but that no cytotoxicity to normal organs or tissues is found. The susceptibility of tumor cells to TRAIL and an apparent lack of activity in normal cells has lead to a proposal to use TRAIL in cancer therapy. Here, we assessed the sensitivity of hepatocytes from rat, mouse, rhesus monkey and human livers to TRAIL-induced apoptosis. TRAIL induced apoptosis in normal human hepatocytes in culture but not in hepatocytes isolated from the other species. Human hepatocytes showed characteristic features of apoptosis, including cytoplasmic shrinkage, the activation of caspases and DNA fragmentation. Apoptosis and cell death in human hepatocytes was massive and rapid, occurring in more than 60% of the cells exposed to TRAIL within 10 hours. These results indicate that there are species differences in sensitivity to TRAIL, and that substantial liver toxicity might result if TRAIL were used in human cancer therapy. PMID- 10802714 TI - Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced hyperglycemia. AB - Insulin gene expression is restricted to islet beta cells of the mammalian pancreas through specific control mechanisms mediated in part by specific transcription factors. The protein encoded by the pancreatic and duodenal homeobox gene 1 (PDX-1) is central in regulating pancreatic development and islet cell function. PDX-1 regulates insulin gene expression and is involved in islet cell-specific expression of various genes. Involvement of PDX-1 in islet-cell differentiation and function has been demonstrated mainly by 'loss-of-function' studies. We used a 'gain-of-function' approach to test whether PDX-1 could endow a non-islet tissue with pancreatic beta-cell characteristics in vivo. Recombinant adenovirus-mediated gene transfer of PDX-1 to the livers of BALB/C and C57BL/6 mice activated expression of the endogenous, otherwise silent, genes for mouse insulin 1 and 2 and prohormone convertase 1/3 (PC 1/3). Expression of PDX-1 resulted in a substantial increase in hepatic immunoreactive insulin content and an increase of 300% in plasma immunoreactive insulin levels, compared with that in mice treated with control adenovirus. Hepatic immunoreactive insulin induced by PDX-1 was processed to mature mouse insulin 1 and 2 and was biologically active; it ameliorated hyperglycemia in diabetic mice treated with streptozotocin. These data indicate the capacity of PDX-1 to reprogram extrapancreatic tissue towards a beta-cell phenotype, may provide a valuable approach for generating 'self' surrogate beta cells, suitable for replacing impaired islet-cell function in diabetics. PMID- 10802715 TI - High susceptibility to bacterial infection, but no liver dysfunction, in mice compromised for hepatocyte NF-kappaB activation. AB - Based on the essential involvement of NF-kappaB in immune and inflammatory responses and its apoptosis-rescue function in normal and malignant cells, inhibitors of this transcription factor are potential therapeutics for the treatment of a wide range of diseases, from bronchial asthma to cancer. Yet, given the essential function of NF-kappaB in the embryonic liver, it is important to determine its necessity in the liver beyond embryogenesis. NF-kappaB is normally retained in the cytoplasm by its inhibitor IkappaB, which is eliminated upon cell stimulation through phosphorylation-dependent ubiquitin degradation. Here, we directed a degradation-resistant IkappaBalpha transgene to mouse hepatocytes in an inducible manner and showed substantial tissue specificity using various means, including a new method for live-animal imaging. Transgene expression resulted in obstruction of NF-kappaB activation, yet produced no signs of liver dysfunction, even when implemented over 15 months. However, the transgene-expressing mice were very vulnerable both to a severe immune challenge and to a systemic bacterial infection. Despite having intact immunocytes and inflammatory cells, these mice were unable to clear Listeria monocytogenes from the liver and succumbed to sepsis. These findings indicate the essential function of the hepatocyte through NF-kappaB activation in certain systemic infections, possibly by coordinating innate immunity in the liver. PMID- 10802716 TI - Cellular immune responses persist and humoral responses decrease two decades after recovery from a single-source outbreak of hepatitis C. AB - As acute hepatitis C virus (HCV) infection is clinically inapparent in most cases, the immunologic correlates of recovery are not well defined. The cellular immune response is thought to contribute to the elimination of HCV-infected cells and a strong HCV-specific T-helper-cell (Th) response is associated with recovery from acute hepatitis C (ref. 2). However, diagnosis of resolved hepatitis C is based at present on the detection of HCV-specific antibodies and the absence of detectable HCV RNA, and detailed comparison of the humoral and cellular immune response has been hampered by the fact that patient cohorts as well as HCV strains are usually heterogeneous and that clinical data from acute-phase and long-term follow-up after infection generally are not available. We studied a cohort of women accidentally exposed to the same HCV strain of known sequence and found that circulating HCV-specific antibodies were undetectable in many patients 18-20 years after recovery, whereas HCV-specific helper and cytotoxic T-cell responses with an interferon (IFN)-gamma-producing (Tc1) phenotype persisted. The data indicate these HCV-specific CD4 + and CD8+ T cells are biomarkers for a prior HCV exposure and recovery. Because of undetectable antibodies against HCV, the incidence of self-limited HCV infections and recovery may be underestimated in the general population. PMID- 10802717 TI - Blockade of interleukin 6 trans signaling suppresses T-cell resistance against apoptosis in chronic intestinal inflammation: evidence in crohn disease and experimental colitis in vivo. AB - The pro-inflammatory cytokine interleukin (IL)-6 (refs. 1-5) can bind to cells lacking the IL-6 receptor (IL-6R) when it forms a complex with the soluble IL-6R (sIL-6R) (trans signaling). Here, we have assessed the contribution of this system to the increased resistance of mucosal T cells against apoptosis in Crohn disease (CD), a chronic inflammatory disease of the gastrointestinal tract. A neutralizing antibody against IL-6R suppressed established experimental colitis in various animal models of CD mediated by type 1 T-helper cells, by inducing apoptosis of lamina propria T cells. Similarly, specific neutralization of sIL-6R in vivo by a newly designed gp130-Fc fusion protein caused suppression of colitis activity and induction of apoptosis, indicating that sIL-6R prevents mucosal T cell apoptosis. In patients with CD, mucosal T cells showed strong evidence for IL-6 trans signaling, with activation of signal transducer and activator of transcription 3, bcl-2 and bcl-xl. Blockade of IL-6 trans signaling caused T-cell apoptosis, indicating that the IL-6-sIL-6R system mediates the resistance of T cells to apoptosis in CD. These data indicate that a pathway of T-cell activation driven by IL-6-sIL-6R contributes to the perpetuation of chronic intestinal inflammation. Specific targeting of this pathway may be a promising new approach for the treatment of CD. PMID- 10802718 TI - The trophoblast is a component of the innate immune system during pregnancy. AB - Systemic infection with Listeria monocytogenes, a Gram-positive intracellular bacterium, has been used extensively to analyze the innate immune response. Macrophages are central to this response, acting as both the host for and principal defense against this bacterium. During pregnancy L. monocytogenes has a predilection for replication at the maternal-placental interface and consequently is an important cause of fetal morbidity and mortality. However, macrophages are mostly excluded from the murine placenta with neutrophils acting as the main immune effector cell against this bacterium. Colony stimulating factor (CSF)-1, a macrophage growth factor, is synthesized in high concentrations by the uterine epithelium during pregnancy, where it is targeted to trophoblast bearing CSF-1 receptors. To define the involvement of CSF-1 in placental immunity, we infected pregnant mice either homozygous or heterozygous for an inactivating recessive mutation in the gene for CSF-1 (osteopetrotic; Csfmop) with L. monocytogenes. CSF 1 was required to recruit neutrophils to the site of listerial infection in the decidua basalis, and infection by Listeria remained unrestrained in its absence. CSF-1 acted by inducing the trophoblast to synthesize the neutrophil chemoattractants (KC) and macrophage inflammatory protein (MIP)-2. Thus, during pregnancy, trophoblast responsive to CSF-1 acts to organize the maternal immune response to bacterial infection at the utero-placental interface. This previously unknown function indicates that the trophoblast acts as a pregnancy-specific component of the innate immune system. PMID- 10802719 TI - A new dual-vector approach to enhance recombinant adeno-associated virus-mediated gene expression through intermolecular cis activation. PMID- 10802721 TI - On the market PMID- 10802720 TI - Overcoming adeno-associated virus vector size limitation through viral DNA heterodimerization. PMID- 10802722 TI - Brave new world view. PMID- 10802723 TI - Single surface stabilizer. PMID- 10802724 TI - Sec1 gets a grip on syntaxin. PMID- 10802725 TI - Compact but disordered states of RNA. PMID- 10802726 TI - New tricks from an itinerant intron. PMID- 10802727 TI - The power of atomic force. PMID- 10802728 TI - Picture story. Four easy pieces. PMID- 10802729 TI - Eukaryotic translation initiation: there are (at least) two sides to every story. AB - The eukaryotic cap and poly(A) tail binding proteins, eIF4E and Pab1p, play important roles in the initiation of protein synthesis. The recent structures of the complex of eIF4E bound to the methylated guanosine (cap) found at the 5'end of messenger RNA (mRNA), the complex of eIF4E bound to peptide fragments of two related translation factors (eIF4G and 4E-BP1), and the complex of the N-terminal fragment of Pab1p bound to polyadenylate RNA have revealed that eIF4E and Pab1p contain at least two distinct functional surfaces. One surface is used for binding mRNA, and the other for binding proteins involved in translation initiation. PMID- 10802730 TI - A collapsed non-native RNA folding state. AB - At physiological Mg2+ concentrations, the catalytic core of the bI5 group I intron does not fold into its native structure. In contrast, as judged by the global size, this RNA undergoes structural collapse at Mg 2+ concentrations much lower than required to drive folding of the RNA completely to the native state. The bI5 RNA therefore exists in equilibrium between expanded and collapsed non native states. The activation energy of RNA folding from the collapsed state to the native state is negligible and the reaction is not accelerated by the addition of urea. This collapsed state is thus distinct from the kinetic traps observed during folding of other large RNAs. The collapsed non-native state forms readily in the case of bI5 RNA and may exist generically prior to assembly of other ribonucleoprotein holoenzymes, such as the ribosome. PMID- 10802731 TI - Small angle X-ray scattering reveals a compact intermediate in RNA folding. AB - We have used small angle X-ray scattering (SAXS) to monitor changes in the overall size and shape of the Tetrahymena ribozyme as it folds. The native ribozyme, formed in the presence of Mg2+, is much more compact and globular than the ensemble of unfolded conformations. Time-resolved measurements show that most of the compaction occurs at least 20-fold faster than the overall folding to the native state, suggesting that a compact intermediate or family of intermediates is formed early and then rearranges in the slow steps that limit the overall folding rate. These results lead to a kinetic folding model in which an initial 'electrostatic collapse' of the RNA is followed by slower rearrangements of elements that are initially mispositioned. PMID- 10802732 TI - Stability and cooperativity of individual tertiary contacts in RNA revealed through chemical denaturation. AB - For proteins, understanding tertiary interactions involved in local versus global unfolding has become increasingly important for understanding the nature of the native state ensemble, the mechanisms of unfolding, and the stability of both the native and intermediate states in folding. In this work we have addressed related questions with respect to RNA structure by combining chemical denaturation and hydroxyl radical footprinting methods. We have determined unfolding isotherms for each of 26 discrete sites of protection located throughout the Tetrahymena thermophila group I ribozyme. The cooperativity of folding, m-value, and the free energy, DeltaG degrees N-U, associated with formation of each tertiary contact was determined by analysis of the isotherms. The DeltaG degrees N-U values measured in this study vary from 1.7 +/- 0.2 to 7. 6 +/- 1.2 kcal mol-1. Thus, the stability of these discrete tertiary contacts vary by almost 104. In addition, an intradomain contact and three interdomain contacts show high cooperativity (m-values of 1.1 +/- 0.2 to 1.7 +/- 0.3 kcal mol-1 M-1) indicating that these contacts exhibit global cooperatively in their folding behavior. This new approach to examining RNA stability provides an exciting comparison to our understanding of protein structure and folding mechanisms. PMID- 10802733 TI - Structural analysis of WW domains and design of a WW prototype. AB - Two new NMR structures of WW domains, the mouse formin binding protein and a putative 84.5 kDa protein from Saccharomyces cerevisiae, show that this domain, only 35 amino acids in length, defines the smallest monomeric triple-stranded antiparallel beta-sheet protein domain that is stable in the absence of disulfide bonds, tightly bound ions or ligands. The structural roles of conserved residues have been studied using site-directed mutagenesis of both wild type domains. Crucial interactions responsible for the stability of the WW structure have been identified. Based on a network of highly conserved long range interactions across the beta-sheet structure that supports the WW fold and on a systematic analysis of conserved residues in the WW family, we have designed a folded prototype WW sequence. PMID- 10802734 TI - Two exposed amino acid residues confer thermostability on a cold shock protein. AB - Thermophilic organisms produce proteins of exceptional stability. To understand protein thermostability at the molecular level we studied a pair of cold shock proteins, one of mesophilic and one of thermophilic origin, by systematic mutagenesis. Although the two proteins differ in sequence at 12 positions, two surface-exposed residues are responsible for the increase in stability of the thermophilic protein (by 15.8 kJ mol-1 at 70 degrees C). 11.5 kJ mol-1 originate from a predominantly electrostatic contribution of Arg 3 and 5.2 kJ mol-1 from hydrophobic interactions of Leu 66 at the carboxy terminus. The mesophilic protein could be converted to a highly thermostable form by changing the Glu residues at positions 3 and 66 to Arg and Leu, respectively. The variation of surface residues may thus provide a simple and powerful approach for increasing the thermostability of a protein. PMID- 10802735 TI - Structure of Cdc42 bound to the GTPase binding domain of PAK. AB - The Rho family GTPases, Cdc42, Rac and Rho, regulate signal transduction pathways via interactions with downstream effector proteins. We report here the solution structure of Cdc42 bound to the GTPase binding domain of alphaPAK, an effector of both Cdc42 and Rac. The structure is compared with those of Cdc42 bound to similar fragments of ACK and WASP, two effector proteins that bind only to Cdc42. The N-termini of all three effector fragments bind in an extended conformation to strand beta2 of Cdc42, and contact helices alpha1 and alpha5. The remaining residues bind to switches I and II of Cdc42, but in a significantly different manner. The structure, together with mutagenesis data, suggests reasons for the specificity of these interactions and provides insight into the mechanism of PAK activation. PMID- 10802736 TI - Structural basis of cell-cell adhesion by NCAM. AB - The neural cell adhesion molecule NCAM, a member of the immunoglobulin superfamily, mediates cell-cell recognition and adhesion via a homophilic interaction. NCAM plays a key role during development and regeneration of the nervous system and is involved in synaptic plasticity associated with memory and learning. The 1.85 A crystal structure of the two N-terminal extracellular domains of NCAM reported here provides a structural basis for the homophilic interaction. The molecular packing of the two-domain structure reveals a cross shaped antiparallel dimer, and provides fundamental insight into trans-cellular recognition mediated by NCAM. PMID- 10802737 TI - Two energetically disparate folding pathways of alpha-lytic protease share a single transition state. AB - The Lysobacter enzymogenes alpha-lytic protease (alphaLP) is synthesized with a 166 amino acid pro region (Pro) that catalyzes the folding of the 198 amino acid protease into its native conformation. An extraordinary feature of this system is the very high energy barrier (DeltaG = 30 kcal mol-1) that effectively prevents alphaLP from folding in the absence of Pro (t1/2 = 1800 years). A pair of mutations has been isolated in the protease that completely suppresses the catalytic defect incurred in Pro by truncation of its last three amino acids. These mutations also accelerate the folding of alphaLP in the absence of Pro by 400-fold. An energetic analysis of the two folding reactions indicates that the mutations stabilize the transition states of both the catalyzed and uncatalyzed folding reactions by 3 kcal mol-1. This finding points to a single transition state for these two distinct and energetically disparate folding pathways, and raises the possibility that all alphaLP folding pathways share the same transition state. PMID- 10802738 TI - RmlC, the third enzyme of dTDP-L-rhamnose pathway, is a new class of epimerase. AB - Deoxythymidine diphosphate (dTDP)-L-rhamnose is the precursor of L-rhamnose, a saccharide required for the virulence of some pathogenic bacteria. dTDP-L rhamnose is synthesized from glucose-1-phosphate and deoxythymidine triphosphate (dTTP) via a pathway involving four distinct enzymes. This pathway does not exist in humans and the enzymes involved in dTDP-L-rhamnose synthesis are potential targets for the design of new therapeutic agents. Here, the crystal structure of dTDP-6-deoxy-D-xylo-4-hexulose 3,5 epimerase (RmlC, EC5.1.3.13) from Salmonella enterica serovar Typhimurium was determined. The third enzyme of the rhamnose biosynthetic pathway, RmlC epimerizes at two carbon centers, the 3 and 5 positions of the sugar ring. The structure was determined by multiwavelength anomalous diffraction to a resolution of 2.17 A. RmlC is a dimer and each monomer is formed mainly from two beta-sheets arranged in a beta-sandwich. The structure of a dTDP-phenol-RmlC complex shows the substrate-binding site to be located between the two beta-sheets; this site is formed from residues of both monomers. Sequence alignments of other RmlC enzymes confirm that this region is very highly conserved. The enzyme is distinct structurally from other epimerases known and thus, is the first example of a new class of carbohydrate epimerase. PMID- 10802739 TI - Voltage dependent activation of potassium channels is coupled to T1 domain structure. AB - The T1 domain, a highly conserved cytoplasmic portion at the N-terminus of the voltage-dependent K+ channel (Kv) alpha-subunit, is responsible for driving and regulating the tetramerization of the alpha-subunits. Here we report the identification of a set of mutations in the T1 domain that alter the gating properties of the Kv channel. Two mutants produce a leftward shift in the activation curve and slow the channel closing rate while a third mutation produces a rightward shift in the activation curve and speeds the channel closing rate. We have determined the crystal structures of T1 domains containing these mutations. Both of the leftward shifting mutants produce similar conformational changes in the putative membrane facing surface of the T1 domain. These results suggest that the structure of the T1 domain in this region is tightly coupled to the channel's gating states. PMID- 10802740 TI - Structure and lipid transport mechanism of a StAR-related domain. AB - The steroidogenic acute regulatory protein (StAR) regulates acute steroidogenesis in the adrenal cortex and gonads by promoting the translocation of cholesterol to the mitochondrial inner membrane where the first step in steriod biosynthesis is catalyzed. StAR-related lipid transfer (START) domains occur in proteins involved in lipid transport and metabolism, signal transduction, and transcriptional regulation. The 2.2 A resolution crystal structure of the START domain of human MLN64 reported here reveals an alpha/beta fold built around a U-shaped incomplete beta-barrel. The interior of the protein encompasses a 26 x 12 x 11 A hydrophobic tunnel that is large enough to bind a single cholesterol molecule. The StAR and MLN64 START domains bind 1 mole of 14C cholesterol per mole of protein in vitro. Based on the START domain structure and cholesterol binding stoichiometry, it is proposed that StAR acts by shuttling cholesterol molecules one at a time through the intermembrane space of the mitochondrion. PMID- 10802741 TI - RNA motifs that determine specificity between a viral replicase and its promoter. AB - The 3' end of brome mosaic virus RNA contains a tRNA-like sequence that directs its RNA synthesis. A stem loop structure in this sequence, stem loop C (SLC), was investigated using NMR, and correlated with its ability to direct RNA synthesis by its replicase. SLC consists of two discrete domains, a flexible stem with an internal loop and a rigid stem containing a 5'-AUA-3' triloop. Efficient RNA synthesis requires the sequence on only one side of the flexible stem and a specific compact conformation of the triloop. A high resolution structure of the triloop places the 5' adenine out in solution, and the 3' adenine within the triloop, held tightly through stacking and unusual hydrogen bonds. This high resolution structure of an RNA promoter from a (+)-strand RNA virus provides new insights into how the RNA-dependent RNA polymerase binds to the RNA to initiate synthesis. PMID- 10802743 TI - Confirmation of the hierarchical folding of RNaseH: a protein engineering study PMID- 10802744 TI - Rational design of potent human transthyretin amyloid disease inhibitors PMID- 10802742 TI - Crystal structure of the Escherichia coli Rob transcription factor in complex with DNA. AB - The Escherichia coli Rob protein is a transcription factor belonging to the AraC/XylS protein family that regulates genes involved in resistance to antibiotics, organic solvents and heavy metals. The genes encoding these proteins are activated by the homologous proteins MarA and SoxS, although the level of activation can vary for the different transcription factors. Here we report a 2.7 A crystal structure of Rob in complex with the micF promoter that reveals an unusual mode of binding to DNA. The Rob-DNA complex differs from the previously reported structure of MarA bound to the mar promoter, in that only one of Rob's dual helix-turn-helix (HTH) motifs engages the major groove of the binding site. Biochemical studies show that sequence specific interactions involving only one of Rob's HTH motifs are sufficient for high affinity binding to DNA. The two different modes of DNA binding seen in crystal structures of Rob and MarA also match the distinctive patterns of DNA protection by AraC at several sites within the pBAD promoter. These and other findings suggest that gene activation by AraC/XylS transcription factors might involve two alternative modes of binding to DNA in different promoter contexts. PMID- 10802745 TI - Protecting the perinatal brain. PMID- 10802746 TI - Mechanisms of perinatal brain injury. AB - This article is focused on the mechanisms underlying primarily ischaemic/reperfusion brain injury in both the term and premature infant. Although the mechanisms involved include similar initiating events, principally ischaemia-reperfusion, and similar final common pathways to cell death, particularly free radical-mediated events, there are certain unique maturational factors influencing the type and pattern of cellular injury. We will therefore initially describe the physiological and cellular/molecular mechanisms of brain injury in the term infant, followed by the mechanisms in the premature infant. PMID- 10802747 TI - Selection of babies for intervention after birth asphyxia. AB - Based on animal experiments, the therapeutic window for neonates with signs of perinatal hypoxia-ischaemia is probably less than 6 h, and early selection of patients is of utmost importance. In term neonates, fetal heart rate and blood flow patterns, the Apgar score, and other clinical scoring systems are insufficient to select patients for intervention, whereas umbilical artery pH<7.0 combined with umbilical arteriovenous differences in PCO(2), lactate/pyruvate ratios in cord blood, and CSF interleukin-1beta have a better predictive value. At present, neurophysiological methods such as (amplitude-integrated) EEG and evoked potentials have the best predictive value. In preterm neonates, lactate/pyruvate and uric acid measurements in cord blood, as well as neurophysiology appear to be helpful to predict brain injury, and might be used to select patients for intervention. PMID- 10802748 TI - Systematic review of therapy after hypoxic-ischaemic brain injury in the perinatal period. AB - Objectives were to identify and to evaluate controlled trials of interventions for term infants developing hypoxic-ischaemic encephalopathy. Five randomized trials concerning prophylactic anticonvulsant therapy for neonatal HIE were identified. There were methodological problems with all of them, and meta analysis of barbiturate prophylaxis showed no significant effect on death or disability. One randomized trial of allopurinol showed short-term benfits, but was too small to test death or disability. One small randomized trial of hypothermia found no adverse effects, but was too small to examine death or disability. No adequate trials of dexamethasone, calcium channel blockers, magnesium sulphate, or naloxone have yet been completed, but pilot studies in infants have shown the risks of magnesium sulphate and calcium channel blockers. PMID- 10802749 TI - Antenatal brain injury: aetiology and possibilities of prevention. AB - Although the aetiology of antenatal brain injury is often unclear, procedures can be employed to prevent or reduce the risk of injury. Defective neuropore closure can be prevented by periconceptional administration of folic acid, and the incidence of other severe malformations and genetic disorders can be reduced by early identification and termination of pregnancy. Antenatal identification of IUGR, administration of corticosteroids to cases with pending preterm birth, and treatment of maternal/fetal infections would also reduce the incidence of injury. Mothers can decrease the risk of injury by maintaining a good diet, avoiding smoking, alcohol intake and exposure to TORCH infections during pregnancy. PMID- 10802750 TI - Intervention after brain injury to reduce disability. AB - After perinatal brain injury, motor function is generally more severely affected than cognition. This article reviews the evidence that intervention after brain injury can reduce disability. There have been few good quality randomized controlled trials. The reasons for this and the difficulties of doing such trials are discussed. The main reasons are: (i) cerebral palsy (CP) is a relatively rare condition; (ii) the patient population is heterogeneous; (iii) different patterns of CP have different prognoses; (iv) a variety of interventions have been used; and (v) outcome measures are relatively poor. Intervention for children considered at risk of developing CP have generally shown no benefit. After children have developed spastic CP, there is a suggestion of some effect due to increasing the frequency of intervention. The precise role of the therapist remains unclear: support of the family may be as important as physical therapy. PMID- 10802751 TI - Cooling the newborn after asphyxia - physiological and experimental background and its clinical use. AB - Many years of experimental work on hypoxic-ischaemic injury have supported the hypothesis that cooling the body and brain after the primary injury offers permanent neuroprotection. Clinically, the question of how late cooling can start after the insult and still have a protective effect is important and not fully investigated. Pilot studies in human adults initiated cooling after 10-18 h (trauma, stroke), however animal data suggest cooling is not effective if started later than 6 h. There might be a threshold for 'cooling dose' - by depth or duration - to achieve permanent protection. Hypothermia must be administered with understanding of the extensive physiological effects. Different enzymes have different sensitivity to changes in temperature, hence some effects may be beneficial and some deleterious. Hypothermia and cardiovascular responses and coagulation needs careful monitoring. PMID- 10802752 TI - Novel treatments after experimental brain injury. AB - Perinatal hypoxic-ischaemic encephalopathy(HIE) is being studied in laboratory models that allow the delayed cascade of events triggered by the energetic insult to be examined in detail. The concept of the 'excitotoxic cascade' provides a conceptual framework for thinking about the pathogenesis of HIE. Major events in the cascade triggered by hypoxia-ischaemia include overstimulation of N-methyl-D aspartate type glutamate receptors, calcium entry into cells, activation of calcium-sensitive enzymes such as nitric oxide synthase, production of oxygen free radicals, injury to mitochondria, leading in turn to necrosis or apoptosis. New experimental approaches to salvaging brain tissue from the effects of HIE include inhibition of neuronal nitric oxide synthase, administration of neuronal growth factors, and inhibition of the caspase enzymes that execute apoptosis. Recent experimental work suggests that these approaches may be effective during a longer 'therapeutic window' after the insult, because they are acting on events that are relatively delayed. Application of modest hypothermia may allow these agents to be neuroprotective at even longer intervals after hypoxia-ischaemia. PMID- 10802753 TI - Distribution and initiation of seizure activity in a rat brain with subcortical band heterotopia. AB - PURPOSE: Misplaced (heterotopic) cortical neurons are a common feature of developmental epilepsies. To better understand seizure disorders associated with cortical heterotopia, the sites of aberrant discharge activity were investigated in vivo and in vitro in a seizure-prone mutant rat (tish) exhibiting subcortical band heterotopia. METHODS: Depth electrode recordings and postmortem assessment of regional c-fos mRNA levels were used to characterize the distribution of aberrant discharge activity during spontaneous seizures in vivo. Electrophysiologic recordings of spontaneous and evoked activity also were performed by using in vitro brain slices from the tish rat treated with proconvulsant drugs (penicillin and 4-aminopyridine). RESULTS: Depth electrode recordings demonstrate that seizure activity begins almost simultaneously in the normotopic and heterotopic areas of the tish neocortex. Spontaneous seizures induce c-fos mRNA in normotopic and heterotopic neocortical areas, and limbic regions. The threshold concentrations of proconvulsant drugs for inducing epileptiform spiking were similar in the normotopic and heterotopic areas of tish brain slices. Manipulations that blocked communication between the normotopic and heterotopic areas of the cortex inhibited spiking in the heterotopic, but not the normotopic, area of the cortex. CONCLUSIONS: These findings indicate that aberrant discharge activity occurs in normotopic and heterotopic areas of the neocortex, and in certain limbic regions during spontaneous seizures in the tish rat. Normotopic neurons are more prone to exhibit epileptiform activity than are heterotopic neurons in the tish cortex, and heterotopic neurons are recruited into spiking by activity initiated in normotopic neurons. The findings indicate that seizures in the tish brain primarily involve telencephalic structures, and suggest that normotopic neurons are responsible for initiating seizures in the dysplastic neocortex. PMID- 10802754 TI - Effects of circadian regulation and rest-activity state on spontaneous seizures in a rat model of limbic epilepsy. AB - PURPOSE: Circadian regulation via the suprachiasmatic nuclei and rest-activity state may influence expression of limbic seizures. METHODS: Male rats (n = 14) were made epileptic by electrical stimulation of the hippocampus, causing limbic status epilepticus and subsequent seizures. We monitored seizures with intrahippocampal electrodes in 12-12-h light/dark (LD) cycles and in continuous dark (DD). We used radiotelemetry monitoring of activity to measure state and body temperature to determine circadian phase. Cosinor analysis and chi2 tests determined whether seizures occurred rhythmically when plotted by phase. State was defined as inactive or active in 10-min epochs based on whether activity count was below or above a cut-off value validated from video observation. RESULTS: In LD, the peak seizure occurrence was 14:59 h after circadian temperature peak (95% confidence limit, 13:37-16:19). Phasic seizure occurrence persisted in DD for 14:05 (12:31-15:38), p < 0.0001, against uniform mean distribution. In LD, 14,787 epochs contained 1, 268 seizures; seizures preferentially occurred during inactive epochs (965 observed, 878 expected in proportion to the overall distribution of inactive versus active epochs; p < 0.001). In DD, 20, 664 epochs contained 1,609 seizures; seizures had no preferential occurrence by state (999 observed, 1,025 expected; p = 0.16). CONCLUSIONS: Limbic seizures occurred with an endogenous circadian rhythm. Seizures preferentially struck during inactivity during entrainment to the light dark cycle. PMID- 10802755 TI - Estradiol facilitates kainic acid-induced, but not flurothyl-induced, behavioral seizure activity in adult female rats. AB - PURPOSE: This study was designed to determine whether previously demonstrated increases in hippocampal axospinous synapse density and NMDA receptor function induced by estradiol are paralleled by increased susceptibility to limbic (kainic acid induced) or generalized (flurothyl induced) behavioral seizures. METHODS: Kainic acid was injected systemically to ovariectomized adult female rats treated with either estradiol or oil vehicle. The latencies to each of five stages of seizure-related behaviors (staring, wet-dog shakes, head waving and chewing, forelimb clonus, rearing, and falling) were recorded for each animal. Flurothyl was administered by inhalation to ovariectomized adult female rats treated with estradiol alone, estradiol followed by short-term progesterone, or oil vehicle. The latencies to each of three stages of seizure-related behaviors (first myoclonic jerk, forelimb clonus, wild running and bouncing) were recorded for each animal. RESULTS: Estradiol treatment decreased the latency to seizure related behaviors induced by kainic acid, but neither estradiol alone nor estradiol followed by progesterone had any effect on flurothyl-induced seizure related behaviors. CONCLUSIONS: The same estradiol treatment paradigm known to induce structural and functional changes in the excitatory circuitry of the hippocampus facilitates the progression of kainic acid-induced seizures, which are known to involve the hippocampus, but has no effect on flurothyl-induced seizures. The lack of an effect of estradiol alone or estradiol followed by progesterone on flurothyl-induced seizures indicates that estradiol's effects on seizure susceptibility do not result from increased neuronal excitability throughout the brain, but rather involve action within the limbic system. The data suggest that structural and functional changes in hippocampal circuitry induced by estradiol may contribute to increased susceptibility to limbic seizure activity. PMID- 10802756 TI - Repeated acute testing of anticonvulsant drugs in amygdala kindled rats: increase in anticonvulsant but decrease in adverse effect potential. AB - PURPOSE: Because preparation of kindled rats is laborious, time-consuming, and expensive, such animals are often used for several experiments in the evaluation of anticonvulsant drugs (AEDs). Furthermore, for comparison with data on new drugs, often "historical" data on standard drugs obtained in previous experiments in other groups of kindled rats are used. Without knowing how factors such as repeated drug testing or seasonal variation in drug responses affect drug potencies in the kindling model, false conclusions and predictions might be drawn from such comparisons. In this study, we examined the anticonvulsant and adverse effects of the three clinically established AEDs carbamazepine (CBZ), phenobarbital (PB), and valproate (VPA) once per month in the same two groups of amygdala-kindled rats over a period of 9 (group 1) or 6 (group 2) consecutive months. To evaluate the possible effect of the season, experiments in group 1 were started in autumn, and experiments in group 2 in spring. METHODS: For quantification of anticonvulsant activity, the focal seizure threshold (threshold for afterdischarges; ADT) was determined after each acute drug treatment and compared with a control ADT determined 2-3 days before. RESULTS: The repeated acute (single-dose) drug testing in the same groups of amygdala-kindled rats led to three pronounced alterations in the animals: (a) a significant decrease in ADT, (b) a marked potentiation of AED effects on ADT, and (c) a striking reduction in ataxia produced by drug treatments. Drug levels in plasma, which were determined in each drug trial, showed only moderate variation over the period of the experiments, so that the observed alterations in drug responses were certainly not due to pharmacokinetic factors. PB and VPA, but not CBZ, showed a more potent anticonvulsant effect when experiments were started in October (group 1) compared with April (group 2), but this difference was rapidly overridden by the marked and progressive potentiation of anticonvulsant activity on repeated drug testing. CONCLUSIONS: These data demonstrate that repeated use of the same kindled rats for acute drug testing significantly alters the sensitivity of the animals to the anticonvulsant and adverse effects of drugs. Because the anticonvulsant potency increases, whereas the adverse effect potential decreases during repeated acute drug testing, this may lead to false positive data on a test compound. The mechanisms involved in these observations deserve further studies. PMID- 10802757 TI - Independent occurrence of the CHRNA4 Ser248Phe mutation in a Norwegian family with nocturnal frontal lobe epilepsy. AB - PURPOSE: To describe the clinical features of a family from Northern Norway in which autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) is associated with a Ser248Phe amino acid exchange in the second transmembrane domain of the neuronal nicotinic acetylcholine receptor alpha4 subunit (CHRNA4). We also tested for evidence of a de novo mutation or founder effect by comparing haplotypes with the original Australian family where the Ser248Phe mutation was first described. METHODS: Clinical details were obtained from 19 family members. Personal interviews and genetic analysis were carried out in 17. Parts of the coding region of CHRNA4 were sequenced, and two known polymorphisms (bp555/FokI, bp594/CfoI) were typed by restriction analysis. RESULTS: Eleven individuals had ADNFLE. The haplotypes of the mutation-carrying alleles of affected individuals from the Northern Norwegian and the Australian ADNFLE family are different. The phenotypic expressions are remarkably similar. CONCLUSIONS: The Ser248Phe mutation occurred independently in both families. Given the rarity of the disease, this suggests that not only the position of a mutation in the coding sequence but also the type of an amino acid exchange is important for the etiology of ADNFLE. The phenotypic similarity of these two families with different genetic backgrounds suggests that the Ser248Phe mutation largely determines the phenotype, with relatively little influence of other background genes. PMID- 10802758 TI - Oxygen desaturations triggered by partial seizures: implications for cardiopulmonary instability in epilepsy. AB - PURPOSE: The occurrence of hypoxemia in adults with partial seizures has not been systematically explored. Our aim was to study in detail the temporal dynamics of this specific type of ictal-associated hypoxemia. METHODS: During long-term video/EEG monitoring (LTM), patients underwent monitoring of oxygen saturation using a digital Spo2 (pulse oximeter) transducer. Six patients (nine seizures) were identified with oxygen desaturations after the onset of partial seizure activity. RESULTS: Complex partial seizures originated from both left and right temporal lobes. Mean seizure duration (+/-SD) was 73 +/- 18 s. Mean Spo2 desaturation duration was 76 +/- 19 s. The onset of oxygen desaturation followed seizure onset with a mean delay of 43 +/- 16 s. Mean (+/-SD) Spo2 nadir was 83 +/ 5% (range, 77-91%), occurring an average of 35 +/- 12 s after the onset of the desaturation. One seizure was associated with prolonged and recurrent Spo2 desaturations. CONCLUSIONS: Partial seizures may be associated with prominent oxygen desaturations. The comparable duration of each seizure and its subsequent desaturation suggests a close mechanistic (possibly causal) relation. Spo2 monitoring provides an added means for seizure detection that may increase LTM yield. These observations also raise the possibility that ictal ventilatory dysfunction could play a role in certain cases of sudden unexpected death in epilepsy in adults with partial seizures. PMID- 10802759 TI - EKG abnormalities during partial seizures in refractory epilepsy. AB - PURPOSE: This study assessed the frequency and character of ictal cardiac rhythm and conduction abnormalities in intractable epilepsy. Sudden unexpected death in epilepsy (SUDEP) is a major cause of excess mortality in people with refractory epilepsy, and cardiac arrhythmias during seizures may be responsible. The frequency of cardiac abnormalities during seizures in patients with refractory epilepsy must be determined. METHODS: Fifty-one seizures in 43 patients with intractable partial epilepsy were analyzed prospectively from CCTV-EEG monitoring with one ECG channel. Arrhythmias, repolarization abnormalities, and PR and QTc intervals were determined for preictal (3 min), ictal, and postictal (3 min) periods for one or more seizures per patient. Parametric statistics were used for continuous variables, and nonparametric statistics were used for categoric variables. RESULTS: Of the patients, 39% had one or more abnormalities of rhythm and/or repolarization during or immediately after seizures. Abnormalities included asystole (one), atrial fibrillation (one), marked or moderate sinus arrhythmia (six), supraventricular tachycardia (one), atrial premature depolarizations (APDs; eight), ventricular premature depolarizations (VPDs; two), and bundle-branch block (three). Mean seizure duration was longer in patients with abnormalities than in those without (204 vs. 71 s; p < 0.001). Generalized tonic-clonic seizures were also associated with increased occurrence of ictal ECG abnormalities (p = 0.006) as compared with complex partial seizures. There were no clinically significant differences in mean preictal and ictal/postictal PR and QTc intervals. CONCLUSIONS: Cardiac rhythm and conduction abnormalities are common during seizures, particularly if they are prolonged or generalized, in intractable epilepsy. These abnormalities may contribute to SUDEP. PMID- 10802760 TI - Epilepsy, vagal nerve stimulation by the NCP system, all-cause mortality, and sudden, unexpected, unexplained death. AB - PURPOSE: This report concerns the 2-year extension of the study of mortality and sudden, unexpected, unexplained death in epilepsy (SUDEP) in the cohort of patients receiving vagal nerve stimulation by the NCP System for the treatment of epilepsy. METHODS: A cohort of 1,819 individuals was followed 3,176.3 person years from implantation. The 25 deaths that occurred during NCP System activation were reviewed for SUDEP by a panel. RESULTS: The mortality rates were lower [standardized mortality ratio (SMR = 3.6)] with the extended follow-up compared to the previous finding (SMR = 5.3). The SUDEP rates (4.1 vs. 4.5 per 1,000 person-years) were similar to those in the previous study of this cohort. When the vagal nerve stimulation experience is stratified by duration of use, the rate of SUDEP was 5.5 per 1,000 over the first 2 years, but only 1.7 per 1,000 thereafter. CONCLUSIONS: The mortality and SUDEP rates are similar to those reported from clinical trials of new drugs and cohorts of severe epilepsy. The lower SUDEP rates after 2 years of follow-up are intriguing, but require further investigation. PMID- 10802761 TI - Risk factors for unsuccessful testing during the intracarotid amobarbital procedure in preadolescent children. AB - PURPOSE: Identification of risk factors for unsuccessful testing during intracarotid amobarbital procedure (IAP) in preadolescent children. METHODS: A pediatric IAP protocol was attempted in 42 candidates for epilepsy surgery (5-12 years old; mean, 10 years) based on the ability to pass a practice test. Language dominance was defined by marked asymmetry until first verbal response and paraphasic errors. Intact hemispheric memory was defined by recall of >/=60% of test items. The odds ratios of baseline variables (age at IAP, Full-Scale IQ, side of disease, age at seizure onset, amobarbital dose) were calculated for various IAP outcomes. RESULTS: IAPs were accomplished in 40 children. Language dominance was established in 25 (62.5%) of 40 patients: all 21 focal resection candidates were left language dominant; four hemispherectomy candidates had intact language after injection of the damaged hemisphere. In 12 (30%) of 40 patients, language testing failed because of agitation or obtundation. Compared with the 21 children with language dominance established by bilateral IAP, these 12 children had lower mean Full-Scale IQ (66.4 vs. 87.9; p = 0.014), and more frequently, the epileptogenic lesion in the left hemisphere (presumed dominant by right-handedness; 78 vs. 33%; p = 0.04). Excluding hemispherectomy candidates (intentionally only one injection), memory testing could not be completed in 13 (36%) of 36 children because of obtundation or agitation. These children were significantly younger than the 23 (64%) of 36 with successful bilateral memory testing (mean age, 107.6 vs. 128.7 months; p = 0. 006). The eight (25%) of 32 children with failing retention scores after ipsilateral injection had lower Full Scale IQ than did the 24 (75%) patients who passed (mean, 59.6 vs. 81.7 months; p = 0.03). CONCLUSIONS: The IAP successfully established hemispheric language dominance and memory representation for just under two thirds of the preselected preadolescent children. Risk factors for unsuccessful testing included low Full Scale IQ (especially <80), young age (especially <10 years), and seizures arising from the left hemisphere presumed dominant by right-handedness. PMID- 10802762 TI - Interictal EEG and ictal scalp EEG propagation are highly predictive of surgical outcome in mesial temporal lobe epilepsy. AB - PURPOSE: Surgical outcome in patients with mesial temporal lobe sclerosis (MTS) is worse than that in patients with temporal lobe activity (TLE) with tumors. Previous studies of the ictal EEG focused on ictal EEG onset in scalp EEG or ictal EEG propagation in invasive recordings. Ictal EEG propagation with scalp electrodes has not been reported. METHODS: Ictal scalp EEG propagation patterns were studied in 347 seizures of 58 patients with MTS or nonlesional TLE. Interictal epileptiform discharges (IEDs) and the presence of unilateral mesial temporal lobe atrophy in magnetic resonance imaging (MRI) also were studied in these 58 patients. Forty-nine patients were operated on (minimal follow-up of 1 year). RESULTS: Postoperatively, seizure-free outcome was seen in (a) 82.8% of patients with regionalized EEG seizure without contralateral propagation, but in only 45.5% of patients with contralateral propagation (p = 0.007); (b) 84.6% of patients with 100% IED lateralized to one temporal lobe, but in only 52.2% with <100% unitemporal IED (p = 0.015); (c) 88.9% with 100% unitemporal IED and regionalized ictal EEG combined, 73.7% with one of both variables, and only 33.3% with <100% ipsitemporal IED combined with contralateral ictal EEG propagation (p = 0.007). CONCLUSIONS: Switch of lateralization or bitemporal asynchrony in the ictal scalp EEG and bitemporal IED are most probably an index of bitemporal epileptogenicity in MTS and are associated with a worse outcome. PMID- 10802763 TI - The role of intracranial electrode reevaluation in epilepsy patients after failed initial invasive monitoring. AB - PURPOSE: Intracranial electrode recording often provides localization of the site of seizure onset to allow epilepsy surgery. In patients whose invasive evaluation fails to localize seizure origin, the utility of further invasive monitoring is unknown. This study was undertaken to explore the hypothesis that a second intracranial investigation is selected patients warrants consideration and can lead to successful epilepsy surgery. METHODS: A series of 110 consecutive patients with partial epilepsy who had undergone intracranial electrode evaluation (by subdural strip, subdural grid, and/or depth electrodes) between February 1992 and October 1998 was retrospectively analyzed. Of these, failed localization of seizure origin was thought to be due to sampling error in 13 patients. Nine of these 13 patients underwent a second intracranial investigation. RESULTS: Reevaluation with intracranial electrodes resulted in satisfactory seizure-onset localization in seven of nine patients, and these seven had epilepsy surgery. Three frontal, two temporal, and one occipital resection as well as one multiple subpial transection were performed. Six patients have become seizure free, and one was not significantly improved. The mean follow-up is 2.8 years. There was no permanent morbidity. CONCLUSIONS: In selected patients in whom invasive monitoring fails to identify the site of seizure origin, reinvestigation with intracranial electrodes can achieve localization of the region of seizure onset and allow successful surgical treatment. PMID- 10802764 TI - Concentric contraction of the visual field in patients with temporal lobe epilepsy and its association with the use of vigabatrin medication. AB - PURPOSE: To describe concentric visual field loss found in the presurgical evaluation of patients with drug-resistant temporal lobe epilepsy and relate the findings to potential causative factors. METHODS: A series of 157 consecutive patients with drug-resistant temporal lobe epilepsy, who had been selected for neurosurgical treatment, was examined in a study set up as a prospective investigation of their visual fields, to document the loss of visual field resulting from surgery. Pre-as well as postoperative visual field examinations were performed following a standard protocol using static and kinetic perimetry. As a number of patients appeared to have an unexplained concentric visual field contraction in the presurgical examination, a relation with potentially causative factors was analyzed in a cross-sectional study of all these patients. Correlations were sought with duration and severity of the seizure disorder, underlying pathology as indicated by magnetic resonance imaging (MRI) and demonstrated by pathology, any type of antiepileptic drug (AED) ever prescribed, and gender. RESULTS: In this cross-sectional analysis of 157 consecutive patients who were candidates for surgery for temporal lobe epilepsy, absolute concentric contraction of the visual field of 10 to 30 degrees was found in the presurgical examination in 20 (17%) of 118 patients who had ever used vigabatrin (VGB) and in none of 39 who had not had this medication. This difference was significant (p = 0.004). In addition, men [15 (21%) of 72] were significantly more often affected (p = 0.007) than women [five (6%) of 85]. The degree of visual field loss, as indicated by the Esterman grid, showed a positive correlation with the duration of VGB medication. There was no correlation of visual field contraction with a history of meningitis as potential cause of the epilepsy, duration of the epilepsy, status epilepticus in the medical history, or histologic abnormality of the brain tissue removed. Ophthalmologic examination of the patients with concentric contraction revealed no abnormalities. None of the patients with concentric contraction complained spontaneously of their visual field loss. CONCLUSIONS: VGB medication is a causative factor in concentric visual field loss. Visual field examination of patients using VGB should be seriously considered. PMID- 10802765 TI - Drug-induced changes in cerebral glucose consumption in bifrontal epilepsy. AB - PURPOSE: Positron emission tomography (PET) using 18F-radiolabeled deoxyglucose (18F-FDG) is a sensitive procedure for detection of epileptogenic foci. Although alterations in glucose consumption are not restricted to the area of seizure generation itself, the magnitude and extent of cerebral metabolic disturbances induced by epileptic discharges can be detected. Despite two decades of epilepsy research using 18F-FDG-PET, little is known about the metabolic changes during therapy of focal epilepsy. We report on a child with frontal epilepsy with severe glucose hypometabolism that was nearly completely normalized during drug therapy. METHODS: Interictal 18F-FDG-PET was performed at the onset of epilepsy and after optimized drug therapy in a 5-year-old boy with behavioral abnormalities and repetitive seizures of frontal origin with bifrontal interictal EEG slowing for 8 weeks. Both scans were anatomically matched; initial and intratherapeutic glucose metabolism were compared. RESULTS: In accordance with the epileptogenic focus as identified by EEG and ictal/interictal perfusion single-photon emission tomography (SPECT), bifrontal hypometabolism was depicted by 18F-FDG-PET. Magnetic resonance imaging (MRI) was unremarkable. After dual-drug therapy (valproate, carbamazepine), the boy became seizure free, and his initial behavioral deficits disappeared. A control PET study after 3 months of therapy showed restored glucose consumption; the frontal EEG slowing was normalized. CONCLUSIONS: This case demonstrates that reduction of glucose metabolism in epileptogenic foci may be a result of reversible neuronal dysfunction that correlates with the electroclinical follow-up. PMID- 10802766 TI - Anterior cingulate gyrus epilepsy: the role of ictal rCBF SPECT in seizure localization. AB - PURPOSE: The goal of this report is to demonstrate the utility of ictal brain single photon emission tomography (SPECT) in a 39-year-old man with complex partial seizures arising from the anterior cingulate gyrus. Seizures originating from the anterior cingulate gyrus are difficult to localize because they have variable ictal semiology, are usually brief, and have rapid cortical propagation. METHODS: Clinical neurologic examination, electroencephalography, extended video electroencephalography with scalp and sphenoidal electrodes, magnetic resonance imaging, computed tomography, and ictal brain SPECT with Tc-99m HMPAO were performed to identify the seizure focus. The patient's regional cerebral blood flow (rCBF) findings were compared with those of eight normal controls, and changes in rCBF were assessed by comparing the patient's ictal scan with those of normal controls at rest by using statistical parametric mapping (SPM). RESULTS: Clinical and neurologic evaluations failed to demonstrate the epileptogenic focus. Ictal rCBF brain SPECT showed a focal region of hyperperfusion in the anterior cingulate gyrus. By using SPM, the ictal blood flow increase in the right anterior cingulate gyrus (x, y, z, -6, 42, 24 mm) was found to be statistically significant when compared with normal controls (z score, 4.88, p < 0.001). Subdural EEG recordings with intracranial electrodes positioned over this location confirmed that the cingulate gyrus was the origin of the seizures, and surgical resection resulted in >90% seizure reduction. CONCLUSIONS: We concluded that ictal brain SPECT localization in conjunction with subdural electrode confirmation is a useful test in the presurgical evaluation of difficult to localize cingulate epilepsy. PMID- 10802767 TI - Acute single photon emission computed tomographic study of vagus nerve stimulation in refractory epilepsy. AB - PURPOSE: Left-sided vagus nerve stimulation (VNS) is an efficacious treatment for patients with refractory epilepsy. The precise mechanism of action remains to be elucidated. Only limited data on VNS-induced changes in regional cerebral blood flow (rCBF) are available. The aim of this study was to investigate rCBF changes during initial VNS with single-photon emission computed tomography (SPECT). METHODS: In 12 patients (8 women, 4 men) with mean age of 32 years and mean duration of epilepsy of 19 years, VNS-induced rCBF changes were studied by means of a 99mTc-ethyl cysteinate dimer activation study with a single-day split-dose protocol before and immediately after initial stimulation. Images were acquired on a triple-head camera with fan-beam collimators and were reconstructed with scatter and attenuation correction. After coregistration to a standardized template, both a semiquantitative analysis using predefined volumes-of-interest (VOIs) as well as voxel-by-voxel analysis of the intrasubject activation were performed. During follow-up, efficacy of VNS in terms of seizure-frequency reduction was studied. RESULTS: The semiquantitative analysis, with reference to the total counts in all VOIs, revealed a significant decrease of activity in the left thalamus immediately after the initial stimulation train. These results agreed with voxel-by-voxel analysis. In our study ipsilateral thalamic hypoperfusion was the most significant finding. Mean frequency of complex partial seizures was reduced from 30 per month before implantation to six per month after implantation. CONCLUSIONS: VNS induces rCBF changes immediately after initial stimulation that can be studied with SPECT. VNS-induced changes in the thalamus may play an important role in suppression of seizures. However, no significant relation between the level of hypoperfusion and subsequent clinical efficacy was found. PMID- 10802768 TI - Cluster analysis of clinical seizure semiology of psychogenic nonepileptic seizures. AB - PURPOSE: To develop an objective classification of psychogenic nonepileptic seizures (NES) based on cluster analysis of clinical seizure semiology. METHODS: We studied the clinical seizure semiology in 27 patients with psychogenic NES documented by prolonged video-EEG monitoring. We analyzed the following clinical symptoms: clonic and hypermotor movements as well as trembling of the upper and/or lower extremities, pelvic thrusting, head movements, tonic posturing backward of the head, and falling. We used cluster analysis to identify symptoms occurring together in a systematic way and thus tried to achieve a clinical classification of psychogenic NES. RESULTS: We could identify three symptom clusters. Cluster 1 was characterized by clonic and hypermotor movements of the extremities, pelvic thrusting, head movements, and tonic posturing of the head, and therefore was named "psychogenic motor seizures." Cluster 2 comprised trembling of the upper and lower extremities and was termed "psychogenic minor motor or trembling seizures." Cluster 3 consisted of falling to the floor as the only symptom and was referred to as "psychogenic atonic seizures." CONCLUSIONS: Our study represents the first study to analyze the clinical semiology of psychogenic NES by cluster analysis, which should be useful for an objective classification of psychogenic NES. This classification should allow both a better characterization of psychogenic NES and an easier differential diagnosis against specific epileptic seizures. PMID- 10802769 TI - Childhood epilepsy and asthma: changes in behavior problems related to gender and change in condition severity. AB - PURPOSE: We conducted a 4-year follow-up study of behavior problems in children with either epilepsy (n = 115) or asthma (n = 105) to identify changes in behavior problems as they were related to gender and change in condition severity. All children were between ages 8 and 13 years and had been diagnosed with their respective conditions for >/=1 year at entry into the study. METHODS: Behavior problems were measured by using the mother's rating on the Child Behavior Checklist. Baseline and follow-up behavior problem scores were examined to see if significant changes occurred over the observation period of the study. To explore change in behavior based on condition severity, each child was placed into "low" and "high" condition severity groups at each time, resulting in four groups: low/low, low/high, high/low, or high/high. There were too few cases in the low/high group to be included in some analyses. Data were analyzed by using analysis of covariance with adjustment for baseline behaviors, age, and age of onset. RESULTS: Within both samples, there was a significant improvement over time for the Total Behavior Problems and Internalizing Problems scores (p /= 1.0 Expanded Disability Status Scale [EDSS] score for an enrollment EDSS score /= 0.5 point for an enrollment EDSS score of >/= 5.5) not associated with an acute relapse. RESULTS: The trial was terminated 1 month after it became fully enrolled due to unanticipated serious cardiopulmonary toxicities (pericarditis, pleural effusion, myocardial infarction, and possible pulmonary embolism), pancreatitis, and death. Notable arthralgia, myalgia, bursitis, and facial and peripheral edema were common adverse events. The high dose of linomide (7.5 mg) was not well tolerated. The trial was too brief to determine unequivocal clinical benefits. Trends suggested an unconfirmed early effect on change in EDSS score at 6 months for the medium dose (2.5 mg daily). CONCLUSION: MS patients may be more prone to develop important linomide treatment-related adverse events than other previously studied patients. However, linomide may be a potentially more toxic drug than was suspected from observations made in smaller studies for other indications. Phase III trials may identify infrequent and important toxicities that may not be anticipated by phase I and II trials. PMID- 10802777 TI - Linomide in relapsing and secondary progressive MS: part II: MRI results. MRI Analysis Center of the University of Texas-Houston, Health Science Center, and the North American Linomide Investigators. AB - OBJECTIVE: To determine the safety and efficacy of roquinimex (linomide) in the management of relapsing-remitting and secondary progressive MS as monitored by MRI. BACKGROUND: Preclinical studies and several short term randomized trials of linomide suggested clinical and MRI-measured benefits with acceptable risk for closely followed MS patients. METHODS: The North American Linomide Trial formally screened 853 individuals for relapsing or secondary progressive, clinically definite MS; recent disease activity or progression; and an Expanded Disability Status Scale score at entry of 3.0 to 6.5 inclusive. MRI was obtained on 811 subjects at pre-enrollment, 718 cases at enrollment, and then at three monthly intervals until the trial was prematurely terminated for unacceptable toxicity. RESULTS: Enhancement was found on 40.2% of 718 entry scans. Statistically robust correlations were found between clinical demographic data and several entry MRI measures including CSF volume, a reflection of brain atrophy. Assessment of the effect of treatment on MRI-measured disease was limited by early trial termination. However, active treatment for 3 months reduced the proportion of patients with one or more enhancements. An exploratory analysis suggested that 2.5 mg was the most active of three doses tested in limiting the total volume of enhanced tissue, the proportion of MRI-defined lesions designated as "black holes," and by a novel MRI composite disease measure. CONCLUSIONS: The short term signature of the effect of linomide on MRI-measured aspects of the disease suggests that safer drugs of this class might be useful in the management of MS. The use of a composite index of the heterogeneous nature of the pathology of MS as captured by MRI may have merit as an outcome measure in clinical trials. PMID- 10802778 TI - Magnetization transfer ratio in new MS lesions before and during therapy with IFNbeta-1a. AB - OBJECTIVE: The authors examined the effect of 6.0 MIU interferon beta-1a (IFNbeta 1a) administered IM each week on the evolution of monthly magnetization transfer ratio (MTR) within new gadolinium-enhancing (Gd+) lesions in patients with very early relapsing-remitting (RR) MS. BACKGROUND: IFNbeta is an effective disease modifying treatment for patients with RRMS. Among other effects, it has been shown to decrease the number of new Gd+ and T2-weighted lesions. MTR is a putative marker for irreversible tissue damage and evolution of MTR within a lesion may reflect recovery of tissue damage. It is not known whether IFNbeta-1a affects the recovery phase of lesions. METHODS: Eight untreated patients with RRMS who completed up to 14 monthly brain MRI sessions elected to initiate treatment with IFNbeta-1a. Four out of eight patients developed new Gd+ lesions during treatment. MTR of lesions at the time of appearance and subsequent rate of change of monthly MTR were compared before and after treatment (stratified Mann Whitney test). RESULTS: The difference between MTR at appearance of 47 new Gd+ lesions before treatment versus 23 new Gd+ lesions during treatment was not significant. Twenty-two of 47 new Gd+ lesions before treatment and 11 of 23 new Gd+ lesions after treatment were monitored for up to 6 months. After appearance of new Gd+ lesions, the rate of increase in MTR was faster during therapy (p = 0.037). CONCLUSION: MTR abnormalities within new Gd+ lesions evolve at a faster rate during treatment with IFNbeta-1a than before initiating therapy. This is consistent with the hypothesis that IFNbeta-1a promotes resolution of new Gd+ lesions. PMID- 10802779 TI - The DYT1 phenotype and guidelines for diagnostic testing. AB - OBJECTIVE: To develop diagnostic testing guidelines for the DYT1 GAG deletion in the Ashkenazi Jewish (AJ) and non-Jewish (NJ) primary torsion dystonia (PTD) populations and to determine the range of dystonic features in affected DYT1 deletion carriers. METHODS: The authors screened 267 individuals with PTD; 170 were clinically ascertained for diagnosis and treatment, 87 were affected family members ascertained for genetic studies, and 10 were clinically and genetically ascertained and included in both groups. We used published primers and PCR amplification across the critical DYT1 region to determine GAG deletion status. Features of dystonia in clinically ascertained (affected) DYT1 GAG deletion carriers and noncarriers were compared to determine a classification scheme that optimized prediction of carriers. The authors assessed the range of clinical features in the genetically ascertained (affected) DYT1 deletion carriers and tested for differences between AJ and NJ patients. RESULTS: The optimal algorithm for classification of clinically ascertained carriers was disease onset before age 24 years in a limb (misclassification, 16.5%; sensitivity, 95%; specificity, 80%). Although application of this classification scheme provided good separation in the AJ group (sensitivity, 96%; specificity, 88%), as well as in the group overall, it was less specific in discriminating NJ carriers from noncarriers (sensitivity, 94%; specificity, 69%). Using age 26 years as the cut-off and any site at onset gave a sensitivity of 100%, but specificity decreased to 54% (63% in AJ and 43% in NJ). Among genetically ascertained carriers, onset up to age 44 years occurred, although the great majority displayed early limb onset. There were no significant differences between AJ and NJ genetically ascertained carriers, except that a higher proportion of NJ carriers had onset in a leg, rather than an arm, and widespread disease. CONCLUSIONS: Diagnostic DYT1 testing in conjunction with genetic counseling is recommended for patients with PTD with onset before age 26 years, as this single criterion detected 100% of clinically ascertained carriers, with specificities of 43% to 63%. Testing patients with onset after age 26 years also may be warranted in those having an affected relative with early onset, as the only carriers we observed with onset at age 26 or later were genetically ascertained relatives of individuals whose symptoms started before age 26 years. PMID- 10802781 TI - Usefulness of MRI measures of entorhinal cortex versus hippocampus in AD. AB - OBJECTIVE: MRI-based measurements of hippocampal atrophy are a sensitive indicator of the early pathologic degeneration of the medial temporal lobe in AD. However, AD pathology appears first in the transentorhinal/entorhinal cortex, not the hippocampus. The authors tested the hypothesis that MRI-based measurements of the entorhinal cortex are more sensitive than measurements of hippocampal volume in discriminating among three clinical groups; controls, patients with a mild cognitive impairment (MCI), and patients with mild probable AD. METHODS: The authors studied 30 controls, 30 patients with MCI, and 30 patients with AD who were matched among clinical groups on age, gender, and education. All underwent a standardized MRI protocol from which the authors made measurements of hippocampal volume, entorhinal cortex volume, and the cumulative length of the medial border of the entorhinal cortex. RESULTS: Pairwise intergroup differences (p < 0.01) were found for all MRI measurements with the exception of the cumulative length of the entorhinal cortex, which did not differentiate controls from MCI patients. Whereas the hippocampal and entorhinal cortex volume measurements provided slightly better intergroup discrimination than the entorhinal distance measurement, overall differences in discriminating ability among the three MRI measurements were minor. CONCLUSIONS: Despite the theoretical rationale for the superiority of entorhinal measurements in early AD, the authors found MRI measurements of the hippocampus and entorhinal cortex were approximately equivalent at intergroup discrimination. Measurements of the hippocampus may be preferable because MRI depiction of the boundaries of the entorhinal cortex can be obscured by anatomic ambiguity, image artifact, or both. PMID- 10802780 TI - Decreased striatal monoaminergic terminals in Huntington disease. AB - OBJECTIVE: To evaluate the integrity of the dorsal striatal dopaminergic innervation in rigid and choreic Huntington disease (HD). BACKGROUND: Some patients with HD have an akinetic-rigid phenotype. It has been suggested that nigrostriatal in addition to striatal pathology is present in this subgroup. The authors sought to determine whether in vivo measures of striatal vesicular monoamine transporter type-2 (VMAT2) binding could distinguish patients with akinetic-rigid (HDr) from typical choreiform (HDc) HD. METHODS: Nineteen patients with HD (mean age 48 +/- 16 years) and 64 normal controls (mean age 50 +/- 14 years) underwent (+)-alpha-[11C]dihydrotetrabenazine (DTBZ) PET imaging. DTBZ blood to brain ligand transport (K1) and tissue to plasma distribution volume (DV) in the caudate nucleus, anterior putamen, and posterior putamen were normalized to the occipital cortex. RESULTS: The normalized striatal specific DV was reduced in HDr (n = 6) when compared with controls: caudate nucleus -33% (p < 0.001), anterior putamen -56% (p < 0.0001), and posterior putamen -75% (p < 0.0001). Patients with HDc (n = 13) also had reduced striatal DV: caudate nucleus -6% (NS), anterior putamen -19% (p < 0.01), and posterior putamen -35% (p < 0.0001). Patients with HDr had significantly lower striatal (+)-alpha-[11C]DTBZ binding than HDc patients. After correction for tissue atrophy effects, normalized DV differences were less significant, with values somewhat increased in the caudate, slightly reduced in the anterior putamen, and moderately decreased in the posterior putamen. There were no significant regional differences in K1 reductions among caudate, anterior, and posterior putamen in HD. CONCLUSIONS: Reduced striatal VMAT2 binding suggests nigrostriatal pathology in HD, most severely in the HDr phenotype. Striatal DV reductions were most prominent in the posterior putamen, similar to PD. PMID- 10802782 TI - PET measures of benzodiazepine receptors in progressive supranuclear palsy. AB - OBJECTIVE: To evaluate the integrity of neurons containing benzodiazepine receptors in metabolically affected regions of the brain in patients with clinically diagnosed progressive supranuclear palsy (PSP). METHODS: The cerebral distribution of [11C]flumazenil (FMZ), a ligand that binds to the gamma aminobutyric acid A (GABAA) receptor, and [18F]fluorodeoxyglucose (FDG), a measure of local cerebral glucose metabolism, was determined with PET in 12 patients with PSP and 10 normal control subjects. Tracer kinetic analysis was applied to quantify data and analysis was performed using three-dimensional stereotactic surface projections and stereotactically determined volumes of interest. RESULTS: There was a global reduction in FMZ binding of 13%, with a reduction in the anterior cingulate gyrus of 20% (p = 0.004), where glucose metabolic rates also showed the greatest reduction. CONCLUSIONS: PSP causes loss of benzodiazepine receptors in the cerebral cortex. Consistent with postmortem studies, the authors did not find significant changes in FMZ binding in subcortical nuclei that exhibit the most pathologic change. This study suggests that both loss of intrinsic neurons containing benzodiazepine receptors and deafferentation of the cerebral cortex from distant brain regions contribute to cerebral cortical hypometabolism in PSP. PMID- 10802783 TI - Predictors of progression in patients with AD and Lewy bodies. AB - OBJECTIVE: To examine the differences in the pattern of progression between AD and AD with Lewy bodies (AD+LB). METHODS: The authors examined predictors of functional and cognitive disability, institutionalization, and death, as well as time to the development of psychosis (e.g., delusions, hallucinations), extrapyramidal signs (EPS), diurnal hypersomnia, and depression in 185 patients with definite AD and 60 with autopsy-confirmed AD+LB. In addition, they analyzed a selected group of patients who did not have comorbid systemic or CNS disease that may have affected progression of the disease (AD = 98 versus AD+LB = 44). The mean follow-up was 58.91 +/- 35.2 months. RESULTS: All cases: Patients with AD+LB had faster time to the development of EPS and diurnal hypersomnia, but not to the development of psychosis or depression. The rate of cognitive and functional decline, time to institutionalization, and physical survival was not different between AD+LB and AD. Selected cases: Patients with AD+LB developed earlier EPS and diurnal hypersomnia than AD patients, and there was a trend to develop earlier major depression, but no differences were noted in time to psychosis. Patients with AD+LB had a faster time to institutionalization than those with AD. The rate of cognitive and functional decline and physical survival was not different between AD+LB and AD in these selected cases. CONCLUSION: Patients with AD+LB can develop EPS and diurnal hypersomnia earlier and have faster time to institutionalization than those with AD alone, but cognitive and functional decline and physical survival are similar between these two entities. PMID- 10802784 TI - Severity of cognitive impairment and the clinical diagnosis of AD with Lewy bodies. AB - OBJECTIVE: 1) To examine the clinical differences between AD and AD with Lewy bodies (AD+LB); and 2) to determine the accuracy of Consensus guidelines for the clinical diagnosis of dementia with Lewy bodies (DLB) at different levels of dementia severity. METHODS: The authors examined the clinical characteristics of 185 patients with pathologically diagnosed AD alone and 60 with AD+LB. The relationship between clinical symptoms and AD+LB was determined by multivariate analyses, controlled by age, duration of symptoms, presence of cerebrovascular disease, and dementia severity. RESULTS: Mild dementia syndrome: No specific clinical symptom was associated with the presence of AD+LB. The sensitivity of the diagnosis of DLB was 62% and specificity was 54%. Moderate dementia syndrome: Extrapyramidal signs (EPS), especially cogwheel rigidity, and major depression were associated with AD+LB. The sensitivity for DLB was 82% and specificity was 31%. Severe dementia syndrome: Cogwheel rigidity and diurnal hypersomnia were associated with AD+LB. The sensitivity for DLB was 93% and specificity was 16%. CONCLUSIONS: The presence of EPS is not useful in differentiating AD+LB from AD in patients with mild dementia. However, as the disease progressed, they emerge as defining features, especially cogwheel rigidity. The accuracy of AD+LB diagnosis varies according the severity of the dementia syndrome. The low sensitivity and specificity in AD+LB patients with mild dementia suggest that in early stages AD+LB patients do not present the clinical characteristics of DLB. By contrast, the high sensitivity and low specificity for the diagnosis of DLB in moderate/severe dementia stages suggests that AD patients can also have characteristic symptoms of DLB. These results indicate that the antemortem diagnosis of AD+LB is difficult in all dementia stages, and better clinical and biologic differentiations of these entities are needed. PMID- 10802785 TI - Two brothers with frontotemporal dementia and parkinsonism with an N279K mutation of the tau gene. AB - OBJECTIVE: To characterize the clinical diagnostic features, neuropathologic phenotype of tau deposition, and subunit structure of tau filaments in patients who had an asparagine-to-lysine substitution at codon 279 (the N279K missense mutation) of the gene for microtubule-associated tau protein. BACKGROUND: The N279K mutation is a causative genetic defect for pallidopontonigral degeneration in an American kindred that presents with frontotemporal dementia (FTD) and parkinsonism. METHODS: The authors analyzed retrospectively the clinical symptoms of two Japanese brothers who carry this mutation. Postmortem neuropathologic and electron microscopic studies, and Western blot analysis of insoluble tau were performed to correlate tau-mediated lesions with neurologic deficits. RESULTS: Both patients exhibited impairment in recent memory, parkinsonism, and corticospinal disturbances in addition to FTD. Parkinsonism in one patient was responsive temporarily to l-dopa. There was intense tau deposition in the medial temporal cortices and upper and lower motor neurons with accompanying corticospinal tract degeneration. Two distinct tau isoforms with four microtubule binding repeats, in hyperphosphorylated forms, were the primary constituents of insoluble tau, which aggregated to the filamentous component, termed "paired tubules," in neurons, oligodendrocytes, and astrocytes. The elemental filaments were hollow tubules measuring 11 to 12 nm in diameter, two of which adhered to each other along their longitudinal axes to form "paired tubules." CONCLUSIONS: Early memory loss and pyramidal signs, which are atypical of FTD, can be presenting symptoms in this disorder. The authors demonstrated that the subunit structure of tau filaments is a pair of hollow tubules despite the prevailing twisted ribbon model. PMID- 10802786 TI - Atrial fibrillation and the risk of cerebral white matter lesions. AB - BACKGROUND: Cerebral white matter lesions are often observed on MRI scans of elderly nondemented and demented persons. Their pathogenesis is not fully understood but cerebral hypoperfusion may be involved. Atrial fibrillation is a common finding in elderly subjects and may lead to a reduced cardiac output with cerebral hypoperfusion. The authors investigated the association between atrial fibrillation and the presence of white matter lesions. METHODS: From 1995 through 1996, the authors randomly sampled 1077 subjects from two ongoing prospective population-based studies. From each participant, an electrocardiogram (ECG) was recorded; atrial fibrillation and left ventricular hypertrophy were diagnosed with a computer program. For one of the two groups (553 subjects), earlier ECGs were available (mean follow-up 4.7 years). All subjects underwent 1.5-T MRI scanning; white matter lesions were separately rated for the periventricular and subcortical regions. RESULTS: The prevalence of atrial fibrillation was 1.9% among subjects younger than 75 years and 5.5% in subjects older than 75 years. The total number of subjects with atrial fibrillation was 28. Subjects with atrial fibrillation had severe periventricular white matter lesions more than twice as often as subjects who did not (RR 2.2; 95% CI 1.0 to 5.2) but had no increased risk of subcortical white matter lesions (RR 1.1; 95% CI 0.4 to 2.6). For seven subjects with atrial fibrillation both at baseline and at follow up, these relative risks were 6.3 (95% CI 1.1 to 37.1) and 0.7 (95% CI 0.1 to 3.7). CONCLUSIONS: Atrial fibrillation is associated with periventricular white matter lesions, but not with subcortical white matter lesions. PMID- 10802787 TI - Ipsilateral hemiparesis after putaminal hemorrhage due to uncrossed pyramidal tract. AB - OBJECTIVE: Previous case reports supported the presence of the uncrossed pyramidal tract in exceptional patients. However, most of these case reports have not fully discussed involvement of the motor cortex controlling the ipsilateral limbs. DESIGN AND METHOD: The authors investigated a 62-year-old man who developed right hemiparesis after right putaminal hemorrhage by using MRI, transcranial magnetic stimulation, functional MRI (fMRI), and sensory evoked potentials. He had moderate weakness including the face, spasticity with brisk deep tendon reflexes and Babinski sign, and impaired vibration and position sense, all on the right side. RESULT: A MRI study showed hemorrhage in the right putamen and the wedge-shaped medulla. A fMRI study during a sequential finger opposition task showed activation in the motor cortex ipsilateral to the finger movements, but not on the contralateral side. Sensory evoked potentials showed cortical response ipsilateral to the side of stimulation. CONCLUSION: The pyramidal tract and the dorsal column-medial lemniscus pathway did not cross in the medulla in this patient. In view of the presence of the abnormal shape in the medulla and congenital scoliosis, a congenital factor might be responsible for the uncrossed pyramidal tract and dorsal column-medial lemniscus in this patient. PMID- 10802788 TI - Poststroke depression and emotional incontinence: correlation with lesion location. AB - OBJECTIVE: To correlate the location of stroke with poststroke depression (PSD) and emotional incontinence (PSEI). METHODS: The authors prospectively studied 148 patients (94 men and 54 women, mean age 62 years) with single, unilateral stroke (126 infarcts and 22 hemorrhages) at 2 to 4 months poststroke with regard to the presence of PSD (using Diagnostic and Statistical Manual of Mental Disorders IV criteria and Beck Depression Inventory) and PSEI. The lesion location was analyzed by CT or MRI. RESULTS: Twenty-seven patients (18%) had PSD and 50 (34%) had PSEI. The presence of PSD and PSEI was not related to the nature, laterality, or size of the lesion. The frequency of PSEI, but not of PSD, was higher in women than in men and in ischemic rather than hemorrhagic stroke (p < 0. 05). Although both PSD and PSEI were related to motor dysfunction and location (anterior versus posterior cortex) of the lesion, location was a stronger determinant for PSD (p < 0.05). The prevalence of PSD/PSEI in each location was 75%/100% in frontal lobe of anterior cerebral artery territory, 50%/0 in temporal lobe, 30%/40% in frontal middle cerebral artery territory, 13%/0 in occipital lobe, 19%/45% in lenticulocapsular area, 11%/16% in thalamus, 16%/53% in pontine base, 36%/55% in medulla, and 0/22% in cerebellum. Parietal and dorsal pontine lesions were not associated with PSD or PSEI. PSEI was more closely associated with lenticulocapsular strokes than was PSD (p < 0.01). CONCLUSION: Development of PSD and PSEI is strongly influenced by lesion location, probably associated with the chemical neuroanatomy related to the frontal/temporal lobe-basal ganglia-ventral brainstem circuitry. Although the lesion distribution is similar, PSEI is more closely related to lenticulocapsular strokes than is PSD. PMID- 10802789 TI - Activate your online subscription PMID- 10802790 TI - Significance of interictal bilateral temporal hypometabolism in temporal lobe epilepsy. AB - OBJECTIVE: To assess the clinical implications and the pathophysiologic determinants of interictal bitemporal hypometabolism (BTH) in temporal lobe epilepsy (TLE) not associated with bilateral MRI abnormalities or intracranial space-occupying lesions. METHODS: The authors compared the clinical, interictal, and ictal EEG, Wada test, and neuropsychology data of 15 patients with intractable complex partial seizures of temporal lobe origin and BTH with those of 13 consecutive patients with unilateral TLE associated with unilateral temporal hypometabolism (UTH) who remained seizure free for more than 3 years after anterior temporal lobectomy. 18F-fluorodeoxyglucose PET scans were analyzed visually and semiquantitatively, and ratios of counts in individual temporal areas to the rest of the cerebrum were compared with the corresponding values from 11 normal control subjects and with the nonepileptogenic hemisphere of the 13 patients with UTH. BTH was defined as more than 2.5 SDs below control values for two or more temporal areas on each side irrespective of any asymmetry. RESULTS: BTH reflected bilateral independent seizure onset in eight patients (53%). The topography of the metabolic depression was not a reliable predictor of epileptogenicity, but involvement of the inferior temporal gyrus was related specifically to ipsilateral seizure onset (70% sensitivity, 100% specificity). In patients with unilateral TLE, contralateral hypometabolism was associated with longer disease duration and worst memory performance during the Wada test, which amounted to global amnesia after ipsilateral injection in three patients, precluding surgical treatment. Contralateral seizure spread in the ictal EEG was significantly faster in patients with BTH. CONCLUSIONS: In TLE, symmetric or asymmetric BTH may signal bilateral independent seizure onset in approximately half the patients, especially when involving the inferior temporal gyrus. Alternatively, it may reflect an advanced stage of the disease process, characterized by a breakdown of the inhibitory mechanisms in the contralateral hemisphere, and secondary memory deficit associated with higher risk of postoperative memory decline. Patients with TLE and BTH but without bilateral MRI changes may still be operated on successfully, but surgical suitability should be proved by comprehensive intracranial EEG studies and Wada test. PMID- 10802791 TI - Quality of life measures in epilepsy: how well can they detect change over time? AB - OBJECTIVE: To evaluate the ability of health-related quality of life (HRQOL) measures to detect change over time in persons with epilepsy. BACKGROUND: The application of HRQOL measures in clinical trials has been limited by a dearth of information regarding their abilities to measure change over time (i.e., their responsiveness). To calculate responsiveness, one must categorize subjects as "changed" or "unchanged" by a priori criteria. METHODS: The authors analyzed data collected at baseline and at 28-week follow-up from an antiepileptic drug trial. Two different criteria for classifying subjects as changed or unchanged-one based on seizure frequency (where changed = attainment of seizure freedom) and one based on self-reported overall condition (where changed = improvement in overall condition)-were used. We compared responsiveness indices for two generic (Short Form [SF]-36 and SF-12) and two epilepsy-targeted (Quality of Life in Epilepsy [QOLIE]-89 and QOLIE-31) HRQOL measures. Two scoring procedures for the SF-36, one based on classic test theory and the other on item response theory (IRT), were compared. RESULTS: Effect sizes of the most responsive HRQOL measures were medium to large. The shorter epilepsy-targeted measure had similar responsiveness indices to those of the longer measure. Epilepsy-targeted measures were consistently more responsive than generic measures under the overall condition criterion, but for the seizure freedom criterion, IRT scoring of the SF-36 yielded responsiveness indices comparable to those of the epilepsy-targeted measures. CONCLUSION: Epilepsy-targeted health-related quality of life measures may be preferable to generic ones in longitudinal studies. Selection of a shorter epilepsy-targeted measure does not compromise responsiveness. Item response theory scoring should be applied to epilepsy-targeted HRQOL measures. PMID- 10802792 TI - CSF insulin-like growth factor-1 in infantile neuronal ceroid lipofuscinosis. AB - BACKGROUND: Infantile neuronal ceroid lipofuscinosis (INCL) is a progressive encephalopathy in which the patients are severely disabled by the age of 3 years. It is characterized by cerebral atrophy, selective loss of cortical neurons, and secondary loss of axons and myelin sheaths of the white matter. INCL has been shown to result from a palmitoyl protein thioesterase deficiency. The authors suggested that insulin-like growth hormones and apoptosis might play a role in the pathogenesis of INCL. METHODS: The authors measured insulin-like growth factor-1 (IGF-1) and IGF binding protein 3 (IGFBP-3) in the CSF of patients with INCL by radioimmunoassay at an early stage when myelin was starting to diminish. RESULTS: The authors found low CSF IGF-1 but normal IGFBP-3 in patients with INCL compared with control subjects. Also, they observed apoptotic cell death in biopsies of INCL patients. CONCLUSIONS: Because the IGF system seems to be important for early brain development, myelination, and neuroprotection, the authors suggest that the pathology in INCL may be associated with low CSF IGF-1. PMID- 10802794 TI - Neuro/Images. Kayser-Fleischer corneal ring. PMID- 10802793 TI - Oral zolmitriptan is effective in the acute treatment of cluster headache. AB - OBJECTIVE: To evaluate the efficacy and tolerability of oral zolmitriptan 5 mg and 10 mg and placebo in cluster headache. METHODS: A multicenter, double-blind, randomized, three-period, crossover, outpatient study. Adult patients received placebo and zolmitriptan 5 mg and 10 mg orally for the acute treatment of episodic or chronic cluster headache. Headache intensity was rated by a five point scale: none, mild, moderate, severe, or very severe. Patients only treated moderate to very severe headaches. The primary efficacy measure was headache response (two-point or greater reduction from baseline in the cluster headache rating scale) at 30 minutes. Secondary efficacy measures included proportion of patients with initial headache relief within 15 and 30 minutes, mild or no pain at 30 minutes, meaningful headache relief, and use of escape medication. RESULTS: A total of 124 patients took at least one dose of study medication, with 73% having episodic and 27% chronic cluster headache. For the primary endpoint, there was a treatment-by-cluster-headache-type interaction (p = 0.0453). Therefore, results are presented separately for chronic and episodic cluster headache. In patients with episodic cluster headache, the difference between zolmitriptan 10 mg and placebo at 30 minutes reached significance (47% versus 29%; p = 0.02). Mild or no pain at 30 minutes was reported by 60%, 57%, and 42% patients treated with zolmitriptan 10 mg, zolmitriptan 5 mg, and placebo (both p left medial temporo-occipital infarct, and a left splenial infarct. TD as an isolated symptom may occur from lesion in a variety of posterior medial locations, including the parahippocampus, splenium, and retrosplenial cortex. PMID- 10802803 TI - Intermanual referral of sensation to anesthetic hands. AB - This report describes a study of patients with hands rendered anesthetic by stroke or neurosurgery. Touching the normal hand of such patients triggers sensations referred contralaterally to the anesthetic hand, paralleling observations of sensory referral to phantom limbs of amputees. The referred somatic sensations are elicited by touch but not usually by other kinds of stimuli, cannot be localized precisely, and do not support spatially organized perception. These characteristics suggest that referral may depend on reorganization in parietal cortical areas other than area 3b, the primary cortical recipient of cutaneous sensory inputs. PMID- 10802805 TI - Obstructive sleep apnea: A study by simultaneous polysomnography and ultrasonic imaging. PMID- 10802804 TI - Migraine with aura and white matter abnormalities: Notch3 mutation. AB - The authors report on an Italian family with eight affected members who show autosomal dominant migraine with prolonged visual, sensory, motor, and aphasic aura. These symptoms are associated with white matter abnormalities on brain MRI. All living affected members carry a Notch3 mutation (Arg153Cys) previously reported in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). White matter abnormalities occur in a variable percentage of the general migraine population; CADASIL should be suspected in migraineurs with prolonged atypical aura and white matter abnormalities. PMID- 10802806 TI - Pyridostigmine-induced microcephaly. PMID- 10802807 TI - Splice site mutation causing a seven amino acid Notch3 in-frame deletion in CADASIL. PMID- 10802808 TI - Decreased CSF amyloid beta42 and normal tau levels in dementia with Lewy bodies. PMID- 10802809 TI - Transderm-induced psychosis in Parkinson's disease. PMID- 10802810 TI - Cardiac uptake of [123I]MIBG separates PD from multiple system atrophy. PMID- 10802811 TI - Cognitive and MRI aspects of corticobasal degeneration and progressive supranuclear palsy. PMID- 10802812 TI - Cerebroretinal vasculopathy mimicking a brain tumor. PMID- 10802816 TI - Special Selection: An Ulcerated Nodule Associated With Lymphadenopathy. PMID- 10802817 TI - Living in Medicine: Heather. PMID- 10802813 TI - Pompholyx after IV immunoglobulin therapy for neurologic disease. PMID- 10802818 TI - JAMA 100 years ago: Medical ethics and medical journals. PMID- 10802820 TI - Quick Uptakes: Spine Strength. PMID- 10802821 TI - Quick Uptakes: Urine Test for Prostate Cancer. PMID- 10802822 TI - Quick Uptakes: Survey of SANE Programs. PMID- 10802823 TI - Quick Uptakes: Geriatrics Guide. PMID- 10802825 TI - Suicidal ideation in adolescent psychiatric inpatients as associated with depression and attachment relationships. AB - Examined suicidal ideation in adolescent psychiatric inpatients as associated with depressive symptoms and attachment to mother, father, and peers. Fifty-nine adolescent psychiatric inpatients (25 male, 34 female) completed self-report measures of suicidal ideation, depressive symptoms, and attachment. Attachment to mother accounted for significant variation in levels of depressive symptoms and suicidal ideation. In addition, attachment to peers accounted for significant variation in suicidal ideation and girls' (but not boys') depressive symptoms. However, after adjusting for depressive symptoms, attachment variables failed to contribute additional variance in suicidal ideation. Self-reported depressive symptomatology remained the strongest predictor of suicidal ideation, regardless of its order in the regression analyses. Prevention and treatment efforts may focus on mother-adolescent attachment and peer attachment (particularly in girls) to reduce risk for depression and suicidal ideation. PMID- 10802826 TI - A test of the hopelessness theory of depression in youth psychiatric inpatients. AB - Evaluated the hopelessness theory of depression among youth psychiatric inpatients. According to the hopelessness theory of depression, negative attributional style may cross-sectionally relate to an array of psychopathological symptoms. However, in the presence but not the absence of negative life events, negative attributional style relates to the onset and exacerbation of depressive (not other) symptoms. Moreover, negative attributional style relates to depression onset or exacerbation specifically via changes in hopelessness (not other mediators). This study of 60 youth psychiatric inpatients (22 boys and 38 girls; ages 9 to 17, M = 14.33, SD = 1.86), 34 of whom were assessed 2 months after leaving the hospital, tested hopelessness theory. At baseline, participants completed self-report questionnaires on attributional style, hopelessness, self-esteem, depression, and anxiety; at follow-up, questionnaires on negative life events and symptoms were completed. Chart diagnoses were available. Results were consistent with all hypotheses derived from hopelessness theory. PMID- 10802827 TI - Why young people do not kill themselves: the reasons for living inventory for adolescents. AB - Assessed the reliability, validity, and predictive power of a new measure, the Reasons for Living Inventory for Adolescents (RFL-A; Osman et al., 1998). A group of 206 (101 boys and 105 girls) adolescent psychiatric inpatients completed the RFL-A, Minnesota Multiphasic Personality Inventory for Adolescents (Butcher et al., 1992), and a packet of self-report measures. Additional information about the patients including diagnosis and suicide status were obtained from their medical records. It was determined that the RFL-A is a valid and reliable measure of adolescent suicide risk potential. Additionally, the RFL-A possesses better predictive power than the Beck Hopelessness Scale (Beck, Weissman, Lester, & Trexler, 1974). A discussion of the clinical and research utility of the RFL-A is included along with suggestions for future research. PMID- 10802828 TI - Psychological, cognitive, and interpersonal correlates of attributional change in adolescents. AB - Examined the role of attributional style in adolescent's psychological functioning. Specifically, we examined the cross-sectional correlates of attributional style, as well as the correlates of changes in attributional style over time. A sample of 841 adolescents with either maladaptive or adaptive attributional styles completed a battery of self-report measures at 2 points in time, 1 year apart. Measures assessed depressive symptoms and suicidality, cognitive functioning (self-esteem, pessimism, coping skills), and interpersonal functioning (social competence, conflict with parents, social support from family and friends). Results indicated that attributional style is associated with multiple depression-related variables. In addition, youth experienced significant changes in their attributional styles over time (from adaptive to maladaptive and vice versa). Finally, changes in attributional style were associated with changes in psychological symptoms and other psychosocial variables. Results are discussed in terms of their implications for the prevention and treatment of adolescent depression. PMID- 10802829 TI - Cognitive specificity in internalizing and externalizing problems in community and clinic-referred children. AB - Examined whether cognitive features in Beck's (1967) model of depression and his cognitive content-specificity hypothesis (Beck, Brown, Steer, Eidelson, & Riskind, 1987) are broadband specific features that distinguish internalizing problems from externalizing problems. From a community (n = 389; 196 boys, 193 girls; age M = 9.98, SD = 1.52) and clinic (n = 82; 55 boys, 27 girls; age M = 10.42, SD = 2.91) sample, 4 groups were defined on behavioral measures and then examined on cognitive measures: externalizing only (n = 33), internalizing only (n = 41), comorbid externalizing and internalizing (n = 53), and control (neither externalizing nor internalizing; n = 35). Both the internalizing only and comorbid groups reported significantly more cognitive disturbances (negative cognitive triad, cognitive processing distortions, and "depressive" and "anxious" thought content) than both of the externalizing only and control groups in both the community and clinic samples. The results were not related to either overall level of psychopathology (as reported by mothers) or social desirability in children's reporting. These findings suggest that the cognitive features in Beck's model distinguish internalizing from externalizing problems and add to a growing literature that calls for further conceptual refinement of cognitive models for understanding both narrow-band and broadband syndromes. PMID- 10802830 TI - Retrospective self-reports of therapist flexibility in a manual-based treatment for youths with anxiety disorders. AB - Evaluated retrospective therapist ratings of the flexibility used when applying the procedures and strategies prescribed in the treatment manual. Flexibility ratings were collected from 18 therapists who had treated 148 children with anxiety disorders (ages 9 through 13). Analyses revealed strong reliabilities for a flexibility questionnaire and that flexibility was used by therapists; however, significant relations between therapist-rated flexibility and treatment outcome were not found. Discussion focuses on the role of flexibility in manual-based psychological treatments and future directions for study. PMID- 10802831 TI - Frequency, comorbidity, and psychosocial impairment of specific phobia in adolescents. AB - Investigated the frequency, comorbidity, and psychosocial impairment of specific phobia and specific fears among 1,035 adolescents 12 to 17 years old. The adolescents were recruited from 36 schools in the province of Bremen, Germany. Specific phobia and other psychiatric disorders were coded based on Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) criteria using the computerized Munich version of the Composite International Diagnostic Interview (Wittchen & Pfister, 1996). Thirty-six (3.5%) of the adolescents met DSM-IV criteria for specific phobia sometime in their life. Of all the subtypes of specific phobia, animal and natural environment phobia were the most common. More girls than boys received the diagnosis of specific phobia. One third of the adolescents with specific phobia also had depressive and somatoform disorders. Despite the high level of psychosocial impairment experienced by individuals with specific phobia both during the worst episode of their disorder and in the last 4 weeks, only a small portion of them sought professional help. PMID- 10802832 TI - The additive and interactive effects of parenting and temperament in predicting adjustment problems of children of divorce. AB - Investigated the interaction between parenting and temperament in predicting adjustment problems in children of divorce. The study utilized a sample of 231 mothers and children, 9 to 12 years old, who had experienced divorce within the previous 2 years. Both mothers' and children's reports on parenting, temperament, and adjustment variables were obtained and combined to create cross-reporter measures of the variables. Parenting and temperament were directly and independently related to outcomes consistent with an additive model of their effects. Significant interactions indicated that parental rejection was more strongly related to adjustment problems for children low in positive emotionality, and inconsistent discipline was more strongly related to adjustment problems for children high in impulsivity. These findings suggest that children who are high in impulsivity may be at greater risk for developing problems, whereas positive emotionality may operate as a protective factor, decreasing the risk of adjustment problems in response to negative parenting. PMID- 10802833 TI - Cognitive interference for trauma cues in sexually abused adolescent girls with posttraumatic stress disorder. AB - Investigated cognitive processing of fear-relevant information in sexually abused adolescent girls with posttraumatic stress disorder (PTSD) using a modified Stroop procedure (MSP). Participants were 20 sexually abused girls with PTSD, 13 sexually abused girls without PTSD, and 20 nonvictimized girls who served as controls, 11 to 17 years old. Word conditions included abuse-related threat, developmentally relevant (related to the experience of sexual abuse, e.g., trust, secrecy, and intimacy), general threat, positive, and neutral. Girls with PTSD were expected to show cognitive interference for trauma-related words as well as for developmentally relevant words, relative to adolescents without PTSD. Overall color naming was significantly slower in the PTSD group than in the nonabused controls. Contrary to expectation, all participants demonstrated cognitive interference for trauma-related words. Relevant theoretical and methodological issues are highlighted. PMID- 10802834 TI - Child sexual abuse prevention programs: a meta-analysis. AB - Conducted a meta-analytic evaluation of the effectiveness of school-based child abuse prevention programs. Literature searches identified 27 studies meeting inclusion criteria for use in this meta-analysis. The average effect size for all programs studied was 1.07, indicating that children who participated in prevention programs performed 1.07 SD higher than control group children on the outcome measures used in the studies. Analysis of moderator variables revealed significant effects for age, number of sessions, participant involvement, type of outcome measure, and use of behavioral skills training. Most important, programs presented over 4 or more sessions that allowed children to become physically involved produced the highest effect sizes. Although most often used only with younger children, findings suggest that active, long-term programs may be more effective for children of all ages. PMID- 10802835 TI - Mother and father self-reports of corporal punishment and severe physical aggression toward clinic-referred youth. AB - Examined the extent to which 359 mothers and 140 fathers of clinic-referred youth (ages 2 to 17) reported using corporal punishment and severe physical aggression when asked directly via intake screening questionnaires at a community mental health center; higher prevalence rates emerged compared to families in the general population. Clinic-referred parents reported greater use of corporal punishment for younger relative to older youth, sons relative to daughters, and by single relative to married mothers. In cases with reports from both parents, mothers used corporal punishment more frequently than fathers. Demographic factors were not linked to severe physical aggression, except for mothers' treatment of sons versus daughters. After controlling for demographic factors, maternal and paternal reports of child externalizing behavior accounted for significant variance in their own and their partner's use of corporal punishment, and in mothers' use of severe physical aggression. PMID- 10802836 TI - The structure of self-reported aggression, drug use, and delinquent behaviors during early adolescence. AB - Examined the structure of self-report scales designed to assess the frequency of adolescent problem behaviors. Urban (n = 988) and rural (n = 1,895) middle school students completed the Problem Behavior Frequency Scale (Farrell, Danish, & Howard, 1992a) and measures of other relevant constructs. Confirmatory factor analyses supported a model that included specific factors related to aggression, drug use, and delinquent behaviors, and a higher order problem behavior factor. Findings did not support a distinction between physical and nonphysical aggression. Results were generally consistent across settings (i.e., urban vs. rural) and gender. Other relevant constructs, including peer pressure for drug use and attitudes favoring aggression, had both specific associations with relevant first-order factors and more general associations with the second-order factor. These findings support the construction of separate scales assessing specific domains of problem behaviors in studies of adolescents' problem behaviors. PMID- 10802837 TI - Then and now. PMID- 10802838 TI - Comparison of a closed (Trach Care MAC) with an open endotracheal suction system in small premature infants. AB - OBJECTIVE: To determine whether ventilated, low birth weight infants treated with closed versus open tracheal suction in a neonatal intensive care unit (NICU) differ as to airway bacterial colonization, nosocomial pneumonia, bloodstream infection (BSI), incidence and severity of bronchopulmonary dysplasia (BPD), neonatal mortality, frequency of suction, reintubation, and nurse preference. STUDY DESIGN: A total of 175 low birth weight infants (< or = 1250 gm) consecutively born (1997 to 1999), intubated, and ventilated in the delivery room were randomized on admission to the NICU to a closed (Trach Care MAC) or open suction group. Closed multi-use catheters were changed daily; open catheters were changed after every use. Two-pass endotracheal suctioning (both groups) was performed every 8 hours or as needed. Side-port connectors were not used; thus open suction required disconnection from ventilators. Tracheal aspirate cultures were obtained on admission and weekly thereafter. Nosocomial BSI (occurring after 48 hours of life) was documented by positive blood cultures. Radiographs taken before, during, and after tracheal aspirate cultures or BSIs were graded using a semiquantitative system for pneumonia and a modified score for BPD. Nurse preference regarding suction method was recorded. RESULTS: Of the original 175 patients, 10 (5 from each group) died and 32 others (16 form each group) were extubated at or before 7 days of life. The study population comprised 67 patients in the closed group and 66 in the open group who were ventilated longer than 1 week. Groups were not statistically different in terms of demographic and clinical characteristics, such as birth weight (837 vs 876 gm), ventilation (27 vs 26 days), and length of stay (49 vs 40 days). Airway colonization with Gram positive cocci occurred in the majority of patients by 2 weeks of life, regardless of group. A total of 39% of infants in the closed group and 44% of infants in the open group became airway colonized with Gram-negative bacilli; differences were statistically significant. No Gram-negative bacilli species was more likely to be associated with either suction. Nosocomial pneumonia was diagnosed in five patients from each group. Nosocomial BSIs occurred in six closed suction infants and five open suction infants. A comparable number of infants in each group developed severe BPD and were discharged from the hospital on oxygen. A total of 28% of closed suction patients and 27% of open suction patients died. Infants in the closed versus open group were suctioned on average 4.4 and 4.1 times per day and were reintubated 9.7 and 8.6 times per 100 ventilator days, respectively. A total of 40 of 44 NICU nurses considered closed suction to be easier to use, less time-consuming, and better tolerated by the patient. CONCLUSIONS: Closed suction obviates the physiological disadvantage of ventilator disconnection without increasing the rate of bacterial airway colonization, frequency of endotracheal suction and reintubation, duration of mechanical ventilation, length of hospitalization, incidence of nosocomial pneumonia, nosocomial BSI, severity of BPD, and neonatal mortality. Although slightly more expensive, closed suction is perceived by nursing staff to be easier, less time-consuming, and better tolerated by small premature infants requiring mechanical ventilation for > or = 1 week. PMID- 10802839 TI - Neonatal outcome when delivery follows a borderline immature lecithin to sphingomyelin ratio. AB - OBJECTIVE: Occasionally, clinicians are presented with a complicated preterm pregnancy where fetal pulmonary maturity testing might be used to help guide management decisions. However, should delivery be allowed if the lecithin to sphingomyelin ratio (L/S ratio) is not quite mature? The incidence of newborn complications after delivery with L/S ratio values of 1.8 and 1.9 is unknown. The purpose of this study was to evaluate the neonatal morbidity and mortality in patients that delivered with these borderline immature results. STUDY DESIGN: All patients who underwent fetal pulmonary maturity testing were prospectively recorded in log books. An L/S ratio of > or = 2.0 was considered mature. Patients with an L/S ratio of 1.8 or 1.9 were considered "borderline immature." These borderline immature cases were evaluated for the gestational age at amniocentesis, the gestational age at delivery, and neonatal outcome. RESULTS: During the 9-year study period, L/S ratio testing was performed on 2038 patients. Of these, 162 preterm patients (7.9%) had an L/S ratio of 1.8 or 1.9 A total of 63 of these 162 patients delivered < 72 hours after the amniocentesis and met study criteria. The pregnancies ranged from 27 to 36 weeks' gestation. There was a 13% incidence (95% confidence interval (CI) of 4% to 30%) of major neonatal morbidity and a 3% incidence (95% CI of 0% to 17%) of neonatal mortality in the 30 pregnancies with an L/S ratio of 1.8. The incidence of major neonatal morbidity was only 3% (95% CI of 0% to 15%) in the 33 patients with an L/S ratio of 1.9, with no cases of mortality (95% CI of 0% to 9%). CONCLUSION: Based on 95% CIs, the data of this study reveal that the maximum risk for major morbidity is < or = 15%, with a mortality risk of < 10% in a preterm newborn delivered with a 1.9 L/S ratio value. The maximum risk is 30% for major morbidity and 17% for mortality in preterm newborns delivered with a 1.8 L/S ratio. This information may help in the decision-making process of whether to deliver or to observe when faced with a borderline immature L/S ratio result in a complicated preterm pregnancy. PMID- 10802840 TI - Short cervical length by ultrasound and cerclage. AB - OBJECTIVE: To determine whether gravidas with short cervical length on endovaginal ultrasound examination, not in preterm labor, who underwent cervical cerclage have better outcomes compared with those with no cerclage. METHODS: This is an observational study in which data were collected prospectively on women who had ultrasound endovaginal cervical length measurement and were not in preterm labor. The subgroup of women who were < or = 26 weeks' at cervical measurement was analyzed separately. Short cervix was defined as < or = 30 mm. After delivery, charts were reviewed for management and outcomes, performed at the discretion of the attending obstetrician. Two study groups were defined: those with cerclage and those with no cerclage. Predictor variables were cerclage and cervical lengths. Outcome variables were birth weight, gestational age at delivery, and neonatal outcomes. Data were analyzed using the chi-squared, Fisher's exact, and Student's t-tests, a p value of < 0.05 was considered to be significant. RESULTS: A total of 85 patients with cervical lengths of < or = 30 mm were identified; of these 43 had cerclage, and 42 did not. The latter had bedrest, tocolytics, or no intervention. Indications for cervical length measurement were similar in both groups, as were age, insurance status, cervical measurements, preterm premature rupture of membranes, and mode of delivery. The mean gestational age at delivery and birth weight in the cerclage group (34.0 +/- 5.4 weeks'; 2530 +/- 905 gm) were greater than in the no cerclage group (32.0 +/- 6.0 weeks', 2084 +/- 1085 gm, p values of < 0.04 and < 0.04, respectively). Analysis for the subgroup of women who were < or = 26 weeks at first measurement revealed similar results. The relative risk for delivering at < 30 weeks' gestation, for incrementally shorter cervices, was less in the cerclage group. CONCLUSION: Cerclage in gravidas with short cervix measured by endovaginal ultrasound, not in preterm labor, may be associated with neonates of greater gestational age and birth weight, with fewer of these parturients delivering before 30 weeks' gestation. A prospective randomized trial of treatment modalities for asymptomatic shortened cervix is needed. PMID- 10802841 TI - Granulocyte-macrophage colony-stimulating factor and interleukin-5 concentrations in premature neonates with eosinophilia. AB - OBJECTIVE: Eosinophilia is common among premature neonates, but little is known about the cytokines responsible for influencing its onset in neonates. In adults and transgenic mice, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-5 (IL-5) influence the development of eosinophilia. We sought to prospectively determine whether these cytokines correlated with the development of eosinophilia in premature infants. STUDY DESIGN: We measured the absolute eosinophil count (AEC) and serial serum concentrations of GM-CSF and IL-5 in premature neonates with eosinophilia. RESULTS: Among 201 premature neonates, 21 had an eosinophil count of > 700/microliter (occurrence, 10.5%). Of these 21 neonates, 4 had mild (700/microliter to 999/microliter), 13 had moderate (1000/microliter to 2999/microliter), and 4 had severe (> 3000/microliter) eosinophilia. No differences in gestational age or age at onset of eosinophilia were observed between the groups, but the duration of eosinophilia was less with mild than with moderate or severe eosinophilia. A total of 20 of the 21 patients had an infection or necrotizing enterocolitis diagnosed within 48 hours of the onset of eosinophilia. In patients with mild and moderate eosinophilia, serial GM CSF and IL-5 concentrations were below the lowest enzyme-linked immunosorbent assay standard. There was no correlation between AEC and GM-CSF or IL-5 concentration in these infants. However, in one patient with severe eosinophilia, two distinct elevations in IL-5 were noted (34.6 and 46.0 pg/ml); each peak occurred 7 to 8 days before a peak in eosinophil count. CONCLUSION: Eosinophilia is relatively common, and in those neonates with the highest AECs, the duration of eosinophilia can last for > 6 weeks. The majority of cases of eosinophilia occurred in temporal proximity to an infectious illness or necrotizing enterocolitis. Serum concentrations of IL-5 were elevated in only one of our patients with severe eosinophilia. PMID- 10802842 TI - Assessment of motor development in very low birth weight infants. AB - OBJECTIVE: The purpose of the current investigation was to describe the fine- and gross-motor acquisitions of very low birth weight (VLBW) infants during their first 12 months without imposing traditional assessment assumptions. STUDY DESIGN: A nonrandomized, prospective study was used for a sample consisting of 89 VLBW infants whose motor development was assessed neonatally and at 4, 8, and 12 months chronological age. Fine- and gross-motor assessment items were selected and adapted from three standardized assessment instruments. RESULTS: The fine- and gross-motor systems of development appeared to be relatively independent of one another before the 8-month assessment. Thus, early fine-motor control appeared to develop almost to the exclusion of gross-motor progress. Once fine motor control was established by 8 months, numerous gross-motor milestones followed and were present at 12 months chronological age. CONCLUSION: Current and previous data continue to define a developmental picture in which VLBW and fullterm infants achieve the same fine- and gross-motor milestones; however, the developmental pathways whereby milestones are achieved, during at least the first 12 months, appear to be different. Accordingly, applying instruments standardized on fullterm infants to the assessment of VLBW babies may not be serving us well on a number of different issues. PMID- 10802843 TI - Cigarette smoking, low birth weight, and preterm births in low-income African American women. AB - OBJECTIVE: To examine effects of light and heavy cigarette smoking on the incidence of low birth weight (LBW) and preterm births in African American women. DESIGN: A total of 1146 low-income African American women participated in a randomized trial to test the effectiveness of nursing intervention in the reduction of preterm and LBW births. This secondary analysis focused on the effects of cigarette smoking on LBW (< 2500 gm) and preterm (< 37 weeks' gestation) births. RESULTS: Cigarette smoking had a significant effect on both LBW and preterm births. When light smokers were compared with nonsmokers, odds ratios were 1.89 (confidence interval (CI) 1.15, 3.13; p = 0.0127) for LBW births and 1.74 (CI 1.00; 3.02; p = 0.0499) for preterm births. When heavy smokers were compared with nonsmokers, odds ratios were 3.03 (CI 1.90, 4.86; p = 0.001) for LBW births and 2.60 (CI 1.55, 4.35; p = 0.0003) for preterm births. CONCLUSION: Cigarette smoking was associated with significantly higher rates of both LBW and preterm births in this sample of African American women. PMID- 10802844 TI - Pulse oximetry: as good as it gets? PMID- 10802845 TI - Improved practical skills of midwives practicing in the Eastern Cape Province of the Republic of South Africa through the study of a self-education manual. AB - OBJECTIVE: Changes in the ability of midwives to perform practical skills, after completion of the Maternal Care Manual of the Perinatal Education Program, were determined. STUDY DESIGN: A prospective, controlled trial in a study town and two control towns. The practical skills of midwives caring for pregnant women in the towns were determined. Subsequently the Maternal Care Manual was studied by midwives in the study town, and the skills of all midwives were subsequently evaluated. RESULTS: A total of 34 midwives in the study town and 39 midwives in the two control towns were studied. The marks showed a significant (p < or = 0.000) improvement in the study town when comparing the pre- and postintervention marks. The mean improvement in the study town was 36.6%. DISCUSSION: The practical skills of midwives improved significantly. The Perinatal Education Program can be implemented with confidence as a distance learning program that is especially suited to the needs of health workers in remote regions. PMID- 10802846 TI - Congenital misalignment of pulmonary vessels and alveolar capillary dysplasia: how to manage a neonatal irreversible lung disease? AB - Congenital misalignment of pulmonary vessels (MPV) with alveolar capillary dysplasia is a rare condition consisting of anomalous veins in bronchovascular bundles, a decreased number of alveolar capillaries, and increased muscularization of pulmonary arterioles. In the literature, infants reported as having such a malformation developed respiratory distress with persistent pulmonary hypertension and ultimately died. We report the case of an infant with MPV and alveolar capillary dysplasia who was unresponsive to maximal cardiorespiratory support, including high-frequency oscillatory ventilation and inhaled nitric oxide; the infant died of pulmonary hemorrhage after 19 days, during venoarterial extracorporeal membrane oxygenation bypass. We conclude that the diagnosis of MPV and alveolar capillary dysplasia should be considered during autopsy of infants who have died of irreversible persistent pulmonary hypertension. If a lung biopsy in infants with prolonged refractory hypoxemia confirms such diagnosis before death, expensive and invasive treatments such as extracorporeal membrane oxygenation could be avoided. PMID- 10802847 TI - Meconium peritonitis. AB - Meconium peritonitis can have a wide range of presentations. This report discusses two cases that have recently appeared in our neonatal intensive care unit. The first report discusses the case of a meconium pseudocyst in a preterm infant. The second case reports on a newborn baby with a healed bowel perforation during the prenatal period. Finally, a brief discussion of meconium peritonitis is also included. PMID- 10802848 TI - Radiology casebook. Patent omphalomesenteric duct. PMID- 10802849 TI - Special imaging casebook. Congenital cystic mesoblastic nephroma. PMID- 10802850 TI - "Bloodless" may not be "harmless"... PMID- 10802851 TI - Microvascular dysfunction in sepsis. AB - The microvascular dysfunction which occurs in sepsis involves all three elements of the microcirculation: arterioles, capillaries, and venules. In sepsis, the arterioles are hyporesponsive to vasoconstrictors and vasodilators. Sepsis also reduces the number of perfused capillaries, thereby impacting on oxygen diffusion to mitochondria. In the venules of some tissues (e.g., mesentery) there is an inflammatory response characterized by neutrophil infiltration and protein leakage. In addition, PMN-endothelial adhesive interactions occur in precapillary microvessels and capillaries in organs, such as, the lung and heart. Thus, all these elements of the microcirculation are involved in the sepsis-induced inflammation. In this review we address emerging views on the mechanisms involved in the microvascular dysfunction induced by sepsis within the framework of these three basic elements of the microcirculatory unit. PMID- 10802852 TI - Quantification of video-taped images in microcirculation research using inexpensive imaging software (Adobe Photoshop). AB - BACKGROUND: Study end-points in microcirculation research are usually video-taped images rather than numeric computer print-outs. Analysis of these video-taped images for the quantification of microcirculatory parameters usually requires computer-based image analysis systems. Most software programs for image analysis are custom-made, expensive, and limited in their applicability to selected parameters and study end-points. METHODS AND RESULT: We demonstrate herein that an inexpensive, commercially available computer software (Adobe Photoshop), run on a Macintosh G3 computer with inbuilt graphic capture board provides versatile, easy to use tools for the quantification of digitized video images. Using images obtained by intravital fluorescence microscopy from the pre- and postischemic muscle microcirculation in the skinfold chamber model in hamsters, Photoshop allows simple and rapid quantification (i) of microvessel diameters, (ii) of the functional capillary density and (iii) of postischemic leakage of FITC-labeled high molecular weight dextran from postcapillary venules. We present evidence of the technical accuracy of the software tools and of a high degree of interobserver reliability. CONCLUSION: Inexpensive commercially available imaging programs (i.e., Adobe Photoshop) provide versatile tools for image analysis with a wide range of potential applications in microcirculation research. PMID- 10802854 TI - Development of an implantable muscle stimulator: measurement of stimulated angiogenesis and poststimulus vessel regression. AB - OBJECTIVE: We developed a lightweight, totally implantable electrical stimulator designed to elicit contraction of skeletal muscle. The stimulator can be programmed to run for different on-off intervals in a given time period in a fully automatic mode. Using the stimulator, angiogenesis was promoted in order to study the rate at which vessel growth and subsequent regression occurs after stimulus removal. METHODS: A fully implanted digital stimulator was designed and fabricated. The stimulator was embedded subcutaneously in the thoracolumbar region of male Sprague-Dawley rats and the electrodes were tunneled under the skin to the common peroneal nerve of the right hind limb. The stimulator elicited muscle contraction in the hind limb at 10 s-1 using square-wave pulses 0.3 ms in duration, evoking contraction of specific muscles for 8 hours/day for 7 days. RESULTS: Chronic stimulation of the skeletal muscles innervated by the common peroneal nerve led to significant increases in blood vessel density in the tibialis anterior (TA; 26%) and the extensor digitorum longus (EDL; 19%) within 7 days. The vessel density remained elevated at 3 days and 7 days poststimulation, but subsequently decreased to control levels by 14 days poststimulation. CONCLUSION: The new stimulator can promote significant increases in vessel density within 7 days, allowing study of both stimulated vessel growth and poststimulus rarefaction. Because of its small size and reliable timing cycles, the stimulator should prove to be a valuable tool in studying these phenomena. PMID- 10802853 TI - Increased sensitivity to the C-X-C chemokine CINC/gro in a model of chronic inflammation. AB - OBJECTIVE: The C-C chemokine MCP-1 elicits significant neutrophil emigration in rats with chronic adjuvant-induced inflammation, but not in naive animals. We examined responses to the C-X-C chemokine CINC/gro to determine whether this class of chemokine elicits altered neutrophil responses during chronic inflammation. METHODS: CINC/gro was superfused over mesenteric venules of naive rats or animals with chronic adjuvant-induced vasculitis. Antibodies were used to characterize adhesive mechanisms. RESULTS: CINC/gro elicited leukocyte transendothelial migration in adjuvant-immunized rats at 100-fold lower concentrations than required to elicit transmigration in naive animals. In both groups, neutrophils constituted > 95% of the leukocytes recruited by CINC/gro. Using in vitro chemotaxis assays, neutrophils from control and adjuvant-immunized rats responded equally to CINC/gro, suggesting differences in migration were not related to neutrophil phenotype. Differences in adhesion molecule usage were noted in vivo. In control animals, CD18 antibodies blocked CINC/gro-induced neutrophil adhesion and emigration. In adjuvant-immunized animals, an alpha 4 integrin antibody reduced adhesion and emigration, while a CD18 antibody selectively inhibited emigration. CONCLUSIONS: This study demonstrates increased sensitivity to a C-X-C chemokine in a model of chronic inflammation, implicates the alpha 4-integrin in neutrophil adhesion, and demonstrates that CD18 mediates leukocyte transendothelial migration independent from firm adhesion. PMID- 10802855 TI - Activation of adenosine A2 alpha receptors inhibits mast cell degranulation and mast cell-dependent vasoconstriction. AB - OBJECTIVE: Adenosine and inosine accumulate in tissue during periods of ischemia and both molecules have been shown to degranulate mast cells in the hamster cheek pouch via activation of an A3 receptor. An A2-mediated inhibitory action of adenosine on mast cell degranulation has also been reported (16), and the objective of this research was to investigate the role of adenosine A2 receptors in modulating inosine-induced mast cell degranulation and subsequent vasoconstriction of microvessels. METHODS: Cheek pouches of the Golden hamster were prepared for in vivo microscopy. Adenosine, inosine, and other agents were applied either globally in the superfusion solution or to selected regions of the tissue by pipette. RESULTS: Micropipette application of 10(-4) M inosine to periarteriolar mast cells caused a vasoconstriction and an associated mast cell degranulation in 71% of the arterioles tested. The average diameter reduction was 29 +/- 5%. To establish a modulatory role for the A2 receptor, low doses of adenosine (100 nM and 10 nM) were applied globally via the superfusion prior to inosine stimulation. This adenosine pretreatment resulted in a decrease in the incidence of the inosine-induced vasoconstriction (17% and 31%), as well as smaller constrictions (0.5 +/- 1% and 7 +/- 3%). Mast cell degranulation was also reduced by pretreatment with adenosine, as evidenced by a decreased number of mast cells exhibiting ruthenium red dye uptake. The inhibitory effect of adenosine could be eliminated by pretreatment with the nonselective A1/A2 antagonist 8-(p-sulfophenyl) theophylline, which restored the inosine-induced responses to control values. To demonstrate that the effect was A2 alpha mediated, vessels were pretreated with the selective A2 alpha agonist 2-[4-(2 carboxyethyl) phenethylamino]-5'-N-ethylcarboxamidoadenosine (CGS21680). Following this treatment, constriction in response to microapplication of inosine (10(-4) M) occurred in only 11% of the vessels tested; the average constriction was reduced to 2 +/- 2% and no mast cell degranulation was observed. CONCLUSIONS: We conclude that mast cell degranulation can be inhibited via activation of an adenosine A2 alpha receptor; which activation occurs at a lower concentration of adenosine than stimulatory A3 receptor activation. This finding may have implications for the pathology of ischemia. PMID- 10802857 TI - Epidemiological and prognostic aspects of gastric MALT-lymphoma. AB - Since mucosa-associated lymphoid tissue (MALT) lymphoma was defined in the mid 1980s as a clinicopathologic entity, many sets of data on pathological, biological and clinical aspects have been generated. In particular, the finding that this process was responding well to antibiotic treatment fueled interest in it and has led to several clinical trials. This overview deals with epidemiological and prognostic aspects and identifies important questions which need to be answered before data from different sources can be compared. Incidence figures of gastric MALT lymphoma vary between countries and parallel the numbers of all non-Hodgkin's lymphoma. The incidence does not parallel the occurrence data of Helicobacter pylori infection. Incidence figures are highly dependent on the definition used for MALT-type primary gastric lymphomas. Several studies show that some prognostic factors are relevant, for instance stage and grade, whereas other factors such as the International Prognostic Index or treatment are not. These studies do not include the recently introduced antibiotic therapy. The inclusion of recent insights in biology and the treatment of gastric MALT lymphomas in prospective clinical studies will soon answer some of the main questions posed. PMID- 10802856 TI - Hypoxia does not activate ATP-sensitive K+ channels in arteriolar muscle cells. AB - OBJECTIVE: To test the hypothesis that hypoxia activates ATP-sensitive K+ (KATP) channels in cremasteric arteriolar muscle cells, resulting in membrane hyperpolarization and inhibition of norepinephrine-induced contraction. METHODS: Arteriolar muscle cells were isolated enzymatically from second- and third-order arterioles that were surgically removed from hamster cremaster muscles. The effects of hypoxia (PO2 = 12-15 mm Hg) were then examined on norepinephrine induced contraction, membrane currents, and membrane potential in these cells at room temperature. Whole-cell currents and membrane potential were recorded using the perforated patch technique. RESULTS: Hypoxia (12-15 mm Hg PO2) reversibly inhibited norepinephrine-induced contraction to 52 +/- 6% of the response in normoxic solutions (156 mm Hg, n = 12 digests, p < 0.05). These effects of hypoxia could be prevented by superfusion of the cells with either solutions containing the KATP channel antagonist glibenclamide (1 microM) or solutions containing 35 mM K+ to reduce the electrochemical gradient for K+ diffusion. Cromakalim, an activator of KATP channels, also inhibited norepinephrine-induced contraction to a similar extent as hypoxia, and in a glibenclamide and 35 mM K(+) sensitive manner. These results are consistent with the KATP channel hypothesis. In contrast, hypoxia had no effect on estimated whole-cell membrane conductance between -40 and -90 mV in voltage-clamp experiments; on holding current measured at -60 mV in cells superfused with 143 mM K+ under voltage-clamp conditions; or on membrane potential in current-clamp experiments, despite positive effects of cromakalim in all three protocols. These electrophysiological data lead to rejection of the hypothesis that hypoxia activates KATP channels. CONCLUSIONS: Hypoxia inhibits norepinephrine-induced contraction of cremasteric arteriolar muscle cells by a mechanism that does not involve KATP channels. It is speculated that the inhibitory effects of glibenclamide and 35 mM K+ on the effects of hypoxia on contraction resulted from depolarization induced by these treatments rather than specific inhibition of KATP channels. PMID- 10802858 TI - Helicobacter pylori in gastric mucosa-associated lymphoid tissue type lymphoma. AB - Infection with Helicobacter pylori triggers the acquisition of gastric mucosa associated lymphoid tissue (MALT) and provides the background for MALT-type lymphoma development. This concept has been supported by a high association of H. pylori infection and MALT-type lymphoma and by the regression of most lymphomas after eradication therapy. In almost all patients with MALT-type lymphoma, serum antibodies to H. pylori were detectable. However, H. pylori was found only in 78% of the patients on histological examination. In addition to other effects, changes in the gastric micro-milieu caused by tumor infiltration of the gastric mucosa may be responsible for the loss of the bacterium. The discrepancy of high seroprevalence and lower histological yield has been already described in other gastric diseases, e.g. atrophic gastritis or gastric carcinoma with extensive destruction of the gastric mucosa. H. pylori strains expressing the CagA protein have been associated with duodenal ulceration and gastric carcinoma. A very high percentage of patients with MALT-type lymphoma is also infected by CagA+ strains of H. pylori as tested by immunoblotting. Antibodies directed to CagA were detectable in the serum as well as in micro-cultured gastric mucosa. Infection with H. pylori may be a precondition for the development of gastric MALT-type lymphoma. In particular, CagA+ strains of H. pylori may, together with additional up to now unknown factors, play a role in the development of gastric MALT-type lymphoma. PMID- 10802859 TI - Tumor biology of mucosa-associated lymphoid tissue lymphomas. AB - Extranodal lymphomas arising at mucosal sites exhibit clinicopathological features that suggest a closer relationship of these tumors to the structure and function of mucosa-associated lymphoid tissue (MALT) than to lymph nodes. The factors that induce MALT in these tissues are operative in early MALT lymphoma development and the progressive independence on T-cell help defines late stages of MALT lymphoma genesis. PMID- 10802860 TI - Histological grading with clinical relevance in gastric mucosa-associated lymphoid tissue (MALT) lymphoma. AB - Treatment choice in gastric mucosa-associated lymphoid tissue (MALT) lymphoma is dependent on the stage and biological rate of progression and transformation as reflected by grade. In pre-treatment, endoscopic biopsy samples, histological and biological criteria to recognize tumor components with a significantly adverse impact on prognosis have to be defined to select patients who may benefit from Helicobacter pylori (H. pylori) eradication as single modality treatment and those who need "classical" anti-cancer therapy. In a consecutive series of 106 patients with gastric MALT-non-Hodgkin's lymphoma (NHL), it was possible to define criteria to differentiate between low-grade and high-grade (transformed) disease. Moreover, within the low-grade group, a category with a diffuse large cell component of 1-10% with or without non-confluent clusters of blasts could be separated with a significantly worse prognosis (10-year disease-specific survival 90% versus 75%). No clinical parameters of known prognostic significance could account for this difference. In a separate series of 19 patients treated with H. pylori eradication, this morphology was strongly related to the chance of reaching complete remission as an independent risk factor. This suggests that it is possible to define criteria in endoscopic biopsy samples to recognize clinically relevant tumor-progression and that these criteria may serve as a guideline in the choice of therapy. PMID- 10802861 TI - Pathology of intestinal lymphomas. AB - The advent of immunohistological and molecular techniques has enabled the comprehensive characterization of many lymphoma entities. Furthermore, it has increased the consensus in lymphoma classification among pathologists. In this review we describe the pathological features of primary intestinal lymphomas classified according to the revised European-American classification of lymphoid neoplasms. The majority of primary intestinal lymphomas are of B-cell lineage and most of these are high-grade tumors. By morphology they may be classified as diffuse large B-cell lymphomas of centroblastic, immunoblastic or plasmablastic type and Burkitt lymphomas. The latter occur predominantly in the terminal ileum and affect children or young adults. Low-grade extra-nodal marginal-zone lymphoma of the mucosa-associated lymphoid tissue (MALT) type and, less frequently, follicular center-cell lymphomas are the low-grade B-cell lymphomas most commonly observed in this region. The first mentioned tumor and its specific intestinal variant, alpha-chain disease or immunoproliferative small intestinal disease are well known for their indolent clinical course. Primary intestinal mantle-cell lymphoma often presents as multiple lymphomatous polyposis and similarly to its node-based equivalent is associated with an unfavorable prognosis. Most primary intestinal T-cell lymphomas display a characteristic immunophenotype, particular histological features with prominent epitheliotropism and are often associated with celiac disease indicating that these tumors form a specific lymphoma type. It has been termed intestinal T-cell lymphoma or enteropathy-type T-cell lymphoma. Clinically, these are aggressive diseases with a high mortality rate. In summary, primary intestinal lymphomas consist of several entities which display distinct clinicopathological features thus confirming the relevance of lymphoma typing. PMID- 10802862 TI - Gastric mucosa-associated lymphoid tissue lymphoma: implications of animal models on pathogenic and therapeutic considerations--mouse models of gastric lymphoma. AB - There are a number of Helicobacter species that will readily colonise the mouse stomach for the duration of the animal's life. They are Helicobacter felis, "Helicobacter heilmannii" and Helicobacter pylori. Early studies on long-term infection of BALB/c mice showed the presence of lesions resembling low-grade gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Because of the suggestion that H. pylori was the cause of these tumors in humans, this phenomenon was studied further as it was reasoned that the Helicobacter-infected mice would provide a valuable model of the human disease. Low-grade gastric MALT lymphomas have been shown to follow infection with all the Helicobacter species listed above. These lesions are indistinguishable from the human disease with the presence of centrocyte-like cells, characteristic lymphoepithelial lesions and glandular destruction. Treatment with antimicrobial therapy results in regression of the lymphomas. There is evidence of progression to high-grade in some animals. The Helicobacter mouse models of lymphoma are likely to provide important information relevant not just to H. pylori-induced lesions in the human, but to antigen-driven tumors in general. PMID- 10802863 TI - Vaccination against Helicobacter pylori. AB - Given the high prevalence of Helicobacter pylori, vaccination has been suggested as a better strategy than widespread use of antibiotics. Despite the fact that natural immunity is not protective, different antigen preparations made from whole cell sonicates, or recombinant proteins have been shown to induce protective immunity when they are delivered with the appropriate adjuvant. Alternative strategies involving antigen delivery by live, attenuated vaccine carriers are also being considered. However, there is still no clear understanding on the mechanisms underlying the vaccine and the need to define reliable immunological markers is now a prerequisite to improving vaccination strategies. PMID- 10802864 TI - Primary gastric non-Hodgkin's lymphoma: requirements for diagnosis and staging. AB - Tumor stage and histological grading (low grade vs. high grade) determine the prognostic outcome of primary gastric mucosa-associated lymphoid tissue (MALT) type non-Hodgkin's lymphoma (NHL). Any diagnostic uncertainty of clinical staging may have potentially important therapeutic implications, especially if a non surgical approach is favored. Diagnostic procedures for evaluation of gastric lymphoma include gastrointestinal endoscopy, endoscopic ultrasound of the upper gastrointestinal tract, conventional abdominal and cervical ultrasound, thoracic and abdominal computed tomography (CT) scans and bone marrow biopsy. In the present overview, the value of clinical diagnostic procedures is discussed with respect to clinical and endoscopic criteria which may be helpful for evaluation of gastric NHL. Furthermore, clinical staging methods such as endoscopic ultrasound are assessed which may allow pre-operative determination of tumor and lymph-node stage. These novel approaches are compared with the gold standard of pathohistological analysis. In conclusion, the diagnostic procedures presented appear to be helpful for establishing an early diagnosis, however, innovative methods such as endoscopic-bioptic mapping of the stomach and ultrasound-guided fine-needle biopsy techniques may improve pre-therapeutical work-up. PMID- 10802865 TI - Gastric lymphomas: aspects of follow-up and after-care. AB - In recent years, new insights into gastric lymphomas and their etiology and pathogenesis have been gained. The predominant role of infection with Helicobacter pylori in the pathogenesis as a pre-malignant condition of a special lymphoma entity [gastric lymphoma of the mucosa-associated lymphoid tissue (MALT) type], has defined new diagnostic procedures and concepts of treatment. Therefore, prognostic factors (e.g. stage of lymphoma, histopathologic grading, resectability etc.) are extremely important for the intensity and efficacy of follow-up and after-care. Surveillance programs in gastric lymphoma include sequelae of surgical resection, chemotherapy and radiotherapy, and the efficacy of follow-up procedures have to be measured by the prevention of tumor relapse in comparison to the intensity of diagnostic procedures. Since lymphoma relapse may occur both as local or disseminated recurrence in 13%-35% of cases, follow-up procedures have to regard both aspects during surveillance. While these follow-up programs are standardized in epithelial tumors (e.g. colon carcinoma), they are not yet established or comparable for each type of gastric lymphoma. Low-grade MALT lymphomas have to be considered as a new lymphoma entity. In addition, new diagnostic procedures (e.g. molecular parameters such as polymerase chain reaction (PCR) for clonality, endosonography, "gastric mapping") have been found to be important parameters for diagnosis and staging of gastric lymphoma and may therefore be relevant for the course of the disease. The definition of "lymphoma cure" and the impact of these procedures as prognostic factors will have to be discussed and may influence the follow-up of gastric lymphoma. PMID- 10802866 TI - Positron emission tomography for detection and staging of malignant lymphoma. AB - FDG-Positron emission tomography (FDG-PET) is an imaging modality using the physiological tracer glucose [modified as 18-fluorine-2-fluorodeoxyglucose (FDG)], whose uptake and metabolism is increased in malignant cells. While exact tumor staging in lymphomatous diseases is the basis for choosing the appropriate treatment strategy, the detection of nodal and extranodal manifestations are a key prerequisite. This study demonstrates that FDG-PET is an efficient, non invasive method for the staging of primary untreated Hodgkin's lymphoma (HD) and non-Hodgkin's lymphoma (NHL). Clinical PET scanning is very useful in staging lymphoma patients and is more accurate than computed tomography (CT) in detecting lesions. PMID- 10802867 TI - Gastrointestinal lymphomas--the Dutch experience. PMID- 10802868 TI - Gastrointestinal lymphomas: the French experience of the Groupe D'etude des Lymphomes Digestifs (GELD). AB - Since 1983, the French Groupe d'Etude des Lymphomes Digestifs (GELD), under the aegis of the Fondation Francaise de Cancerologie Digestive, has aimed to identify the different prognostic groups of the primary digestive-tract lymphomas (PDTL) and their optimal treatment. Successive multicenter studies were conducted and 91 PDTL were evaluated. A marked improvement in their prognosis was obtained by a strategy including precise histologic typing and clinical staging followed by a therapeutic approach combining initial surgical resection, whenever possible or reasonable, followed by chemotherapy adapted to the grade of malignancy and resectability of the lymphoma. The multivariate analysis indicated that the factors for good prognosis were age (< 65 yrs), gastric localisation, stage IE and radical or even incomplete surgery. However, Helicobacter pylori eradication should be the first treatment in stage IE low-grade gastric mucosa-associated lymphoid tissue (MALT) tumors. The long-term results of such medical treatment are evaluated together with the management and the place of surgery in these localised tumors. However, owing to the limited number of patients, a large international co-operative trial is needed to confirm the findings. Thirty-one cases of multiple lymphomatous polyposis were also collected and confirmed to be a distinct entity among PDTL and the gastrointestinal counterpart of the mantle cell-zone lymphomas. High-dose radio-chemotherapy supported by auto transplantation improved their prognosis. PMID- 10802869 TI - Treatment of primary gastric lymphoma: experience in the National Cancer Center Hospital, Tokyo. AB - The single institutional experience of the treatment of primary gastric lymphoma is presented in chronological order. Between 1963 and 1986, 74 patients were treated with various uncontrolled methods and resection line involvement was seen in seven cases. Between 1987 and 1995, a prospective study was conducted employing total gastrectomy with systematic lymphadenectomy, followed by chemotherapy for cases with lymph node metastasis. Fifty patients were enrolled and the 5-year survival rate was 86%. Thorough histological examinations of the resected specimens revealed multiple foci in the stomach and nodal involvement in 36% and 50% of cases, respectively. Since 1995, the effects of eradication of Helicobactor pylori have been examined in association with the introduction of the histological diagnosis of mucosa-associated lymphoid tissue (MALT) lymphoma. Special attention should be paid to the elevated type tumors because they could metastasize to lymph nodes preserving the features of "low-grade" MALT lymphoma. PMID- 10802870 TI - Primary gastric lymphoma: brief overview of the recent Princess Margaret Hospital experience. AB - Primary gastric lymphoma is the commonest form of presentation for gastrointestinal lymphomas and the stomach is one of the most frequent sites of extranodal lymphoma. We present a review of the Princess Margaret Hospital (PMH) experience to illustrate the favorable prognosis and examine the long-term outcomes in this disease. Between 1967 and 1996, 149 consecutive patients with gastric lymphoma were treated at the PMH. The majority (122 patients) presented with diffuse large-cell lymphoma and 78 had stage I disease. In the past, the standard treatment was surgery (partial gastrectomy) followed by post-operative radiation therapy (RT; 78 patients). The overall 10-year survival was 62%. For patients who were treated with surgery and post-operative RT (operable disease) between 1967 and 1985, the 10-year rates of overall survival and cause-specific survival were 66.2% and 88%, respectively. In the past decade, combined-modality therapy with chemotherapy followed by RT was introduced for large-cell lymphoma, while RT alone was used for mucosa-associated lymphoid tissue (MALT) lymphoma. In 45 patients treated in the past decade, the overall 5-year survival was 86%, the cause-specific survival was 95.5%, and the relapse-free rate was 87.3%. These results support the view that primary gastric lymphoma is a highly curable disease. Future efforts should focus on reducing treatment morbidity, while preserving excellent results. PMID- 10802872 TI - Eradication of Helicobacter pylori and stability of remissions in low-grade gastric B-cell lymphomas of the mucosa-associated lymphoid tissue: results of an ongoing multicenter trial. AB - The normal human stomach is devoid of any organized lymphatic tissue. Acquisition of mucosa-associated lymphatoid tissue (MALT) in the stomach is considered to be a direct consequence of chronic infection with Helicobacter pylori. Thus, MALT appears to be part of the host defense against the pathogen H. pylori. Consequently, lymphomas arising from gastric MALT may be seen as an end point of a clonal evolution starting from the infection. Cumulative data from several studies show that eradication of H. pylori induces complete histologic remissions in about 70%-80% of the patients. Here we present data of an extended analysis of an ongoing multicenter trial. Eighty-four patients with low-grade gastric MALT lymphoma in stage EI were treated using a dual regimen to eradicate H. pylori. Complete remission was observed in 68 (81%) patients; a partial remission was found in seven (8%) patients. In contrast, nine (11%) patients revealed "no change" and were referred for alternative treatment strategies. The majority of these cases were found to harbor high-grade lymphomas in deeper mucosal areas. Polymerase chain reaction (PCR) performed on the VDJ rearrangements of the immunoglobulin heavy chain yielded monoclonal bands in 50 of 65 analyzed patients (77%) at diagnosis. Interestingly, in patients analyzed during follow up after achieving complete histologic remission, ongoing PCR monoclonality was found in 19 of 39 eligible patients (49%). Several patients who developed local relapse of the lymphoma were found in the group with ongoing PCR monoclonality. Together with data from the literature, these results suggest that the majority of low grade gastric MALT lymphomas in stage EI respond to eradication of H. pylori. Longer follow-up investigations are necessary to determine whether remissions really indicate a cure from the disease and to elucidate whether PCR monoclonality after complete histological remission is predictive of increased relapse rate. PMID- 10802871 TI - Relapse of low-grade gastric MALT lymphoma after Helicobacter pylori eradication: true relapse or persistence? Long-term post-treatment follow-up of a multicenter trial in the north-east of Italy and evaluation of the diagnostic protocol's adequacy. AB - The effect of eradication of Helicobacter pylori on early stage gastric low-grade MALT lymphoma in 76 patients with follow-up of at least 1 year (12-63 months, mean 28) is reported. No regression was found in five cases after 12-48 months. In one case surgical resection detected the involvement of perigastric lymph nodes overlooked by endoscopic ultrasonography (EUS). Neither progression of the disease nor a high-grade component was documented by repeated gastric mappings, EUS and complete stagings in the other four cases. After histological remission five relapses of low-grade and one relapse of high-grade MALT lymphoma were found 12-48 months after eradication. Subsequent histological remission, without any additional therapy, was found in three relapsed cases. A rapid and persistent histological remission was obtained in 56 patients (73%). A late remission was observed in six cases. Monoclonal remission was found in half of the patients and was frequently delayed. Persistent monoclonality was associated with histological remission in the vast majority of patients. Our data confirm H. pylori eradication as the first choice therapy for early stage gastric low-grade MALT lymphoma and recommend extensive bioptic mapping and endoscopic sonography both in the local staging and in the regression evaluation. The rare cases of late remission encourage us to wait for at least 1 year after eradication of H. pylori. Longer follow-up studies will clarify the meaning of histological relapse/persistence and late remission. The study of non-responder cases could show us a step in lymphomagenesis. PMID- 10802873 TI - Gastrointestinal lymphomas: the Wurzburg study experience. AB - Appropriate management of primary gastric lymphomas is still controversial. We, therefore, conducted a prospective multicenter study to identify its clinical features, evaluate the accuracy of diagnostic and staging procedures, and assess a treatment strategy based on tumor stage and malignancy. Of 266 patients recruited within three years, 107 had low-grade (40%) and 159 high-grade (60%) lymphoma. A total of 237 patients (89%) presented with localized disease (stages EI/II). Based on the rapid urease test and/or histology the overall Helicobacter pylori positivity was 59% (76%, 51%, 38% in low-grade, high-grade, and secondary high-grade lymphoma, respectively). In 27% of the cases, patients could not be precisely classified and graded on the basis of endoscopic biopsies. In 78 patients, pre-operative endoscopic ultrasound correctly predicted the depth of tumor infiltration in 78% and lymph-node involvement in 75%. Treatment was stratified according to the grade of malignancy and stage: H. pylori eradication in low-grade lymphoma of stage EI, surgical resection in stages EI/II of low- and high-grade lymphoma and, depending on the pathohistological stage and post operative residual tumor mass radiation and chemotherapy/combined radiochemotherapy in low-grade and high-grade lymphoma, respectively. PMID- 10802874 TI - [Clinical and economic evaluation of surgery in the treatment of obstructive sleep apnea syndrome]. PMID- 10802875 TI - The "traditional" American family: implications for marketing. AB - The American family has undergone a major transition over the past several decades. While the relative importance of married couples predominates, there has been a steady increase in the proportion of adults who are divorced, and widowhood continues to increase in line with the evergrowing aged U.S. population. Among the changes in the family structure are a declining role for two-parent families with children more single-parent households and an increasing role for grandparents in many family situations. Within the two-parent family there are changes as well with more working mothers, an increase in women householders and, in some cases, more involvement in the child-rearing process on the part of the male parent. The traditional two-parent family, nevertheless, retains considerable economic clout, benefiting from two working marriage partners. PMID- 10802876 TI - The Social Security system: benefit differences among racial/ethnic groups. AB - Based on published and unpublished data from the Bureau of the Census and the Social Security Administration, an assessment is presented of how major racial/ethnic groups in the United States fare under Social Security. The Social Security Act was signed into law in 1935 to provide economic security to U.S. workers. Today, there are some 44.5 million Social Security beneficiaries in this country. The majority of these beneficiaries are retired workers (62.3 percent) with survivors of deceased workers comprising another 11.6 percent. Several aspects of the Social Security system are particularly important to minority groups. The data indicate that minorities rely more on Social Security benefits in retirement than do whites. In comparison with only 16 percent of white elderly beneficiaries, one-third of elderly blacks and one-third of elderly Hispanics depend on Social Security payments for 100 percent of their retirement income. Minorities also have less retirement income from other sources, such as pensions and assets, than do whites. Both blacks and Hispanics tend to have lower earnings than whites and, thus, benefit from the progressive benefit formula of the Social Security system. Further, blacks, with shorter life expectancies, benefit more from the disability and survivors benefits than do whites. Social Security has a major impact on poverty rates, particularly for minorities. Without Social Security benefits and no other changes in savings or pensions, poverty rates would be around 60 percent for blacks, Native Americans and Hispanics versus 24, 12 and 20 percent, respectively, with Social Security benefits. The U.S. minority population will continue to grow in both numbers and proportions. By 2050, almost half (47 percent) of the total U.S. population will be minorities due principally to growth in Hispanic, black and Asian populations. The increasingly diverse population will change the composition of the nation's workforce as well as the size and makeup of its elderly population. These demographic changes will be important to consider when evaluating the future role that Social Security will play in the lives of these changing populations. PMID- 10802877 TI - Trends in unintentional-injury deaths during the 20th century. AB - Since 1900 the number of deaths per 100,000 population due to unintentional injuries (accidents) was reduced by 53 percent, dropping from 72 to 34. Had this rate not decreased almost 4.2 million more people would have died from unintentional injuries over the century. Of the seven kinds of injury events that accounted for 90 percent of all unintentional injury deaths in 1998, only two experienced increases in rates. Death rates from falls, fires and burns, drowning, poisoning by gases and vapors and firearms were reduced between 61 and 90 percent. The mortality rate due to poisoning by solids and liquids, however, increased 19 percent while that from motor-vehicle crashes sky-rocketed by 7,500 percent. In 1910 there were 468,500 vehicles registered in the United States and the death rate from these crashes was 0.2 per 100,000. In 1998, however, there were 215.4 million vehicles and a death rate of 15.2. Deaths due to solid and liquid poisonings ranged between 1,500 and 2,500 until the late 1960s and now number more than 8,000 annually. The increase is greatest among persons between the ages of 25 and 44 and is largely attributed to illegal drugs. Death rates in this category were lowest in the 1950s and now are as high as in the mid-1910s. Deaths due to falls fluctuated from about 7,400 in 1910 to nearly 25,000 annually in the 1940s, to about 11,400 in 1986, and to 16,600 in 1998. Death rates for age groups under 65 years are at their lowest while those for the 65 and older age group have been increasing. Drowning and firearms deaths have decreased fairly steadily since the early 1970s. Death rates for all age groups have shown steady reductions over the century and are now at or near their lowest points. The firearms death rate for 15-24 year olds, however, was and is substantially greater than all other age groups. The number of deaths due to fires and burns and gas and vapor poisonings have been declining since the mid-1960s and late 1970s respectively. Fire and burn death rates among young children and the elderly have shown the greatest improvement. PMID- 10802878 TI - Average charges for three types of hysterectomy procedures: United States, 1998. AB - The average charges associated with three forms of hysterectomies (abdominal laparotomy) vaginal and laparoscopically-assisted vaginal hysterectomy (LAVH) in 1998 were investigated by geographic area and individual states. Considerable variation in the rates of and charges for these three procedures was reported. Based on data from more than 400,000 women insured under group health contracts, the average charges for these three gynecological surgeries were calculated for the study group of 14,184. The majority of the surgeries were laparotomies (64 percent) with LAVH accounting for only 10 percent; the largest proportion of the procedures were performed in the West South Central and South Atlantic regions of the country and the highest average charge was associated with the LAVH procedures. In 1998, the average charge for an abdominal hysterectomy in the United States was $12,500: that for a vaginal hysterectomy was $10,380; and that for a laparoscopically-assisted vaginal hysterectomy was $14,500. The Pacific area registered the highest average charges for all three (between 19 and 21 percent higher than the U.S. norm) while the lowest geographic area charges were reported in West North Central states (between 18 and 21 percent lower than the U.S. average). The charges in California were the highest of all study states for each of the surgeries, ranging from 38 to 43 percent higher than the average, whereas the charges in Iowa were the lowest for the laparotomies and vaginal hysterectomies (42 and 36 percent, respectively below the norm) and in Kansas for the LAVHs (34 percent lower than the U.S. average). Physicians fees were the highest in New York for each procedure ranging from 33 percent higher than the norm for a vaginal hysterectomy, 50 percent higher for an LAVH and 72 percent higher for a laparotomy. The lowest physician charges were reported in Iowa for the vaginal and abdominal surgeries and in Minnesota for the LAVHs. The average length of stay was 3.10 days for the laparotomy procedures, 2.20 days for the vaginal hysterectomies and 1.99 days for the LAVH patients, with substantial variation between states for each procedure. PMID- 10802879 TI - U.S. population changes since 1900. PMID- 10802880 TI - [Bioinformatics , genomics, proteomics--life sciences of XXI century]. PMID- 10802881 TI - [Processes of modification of lipoproteins. Physiological and pathogenetical role of modified lipoproteins]. AB - The review is devoted to modification of lipoproteins (Lp) connected with various biological processes. Possible mechanisms of Lp oxidation and the nature of oxidants involved in to these processes are considered. Specific attention is paid to the participation of proteolytic enzymes in modification of Lp. PMID- 10802882 TI - [The selenium haemostasis during experimental anaphylaxis reaction in rats treated with reduced glutathione and selenium enriched spirulina]. AB - The main events caused by anaphilaxis in selenium haemostasis in rats include significant increase of selenium excretion with urine (6.36 +/- 1.18 nM Se/18 h., n = 10, compared with 1.72 +/- 0.38 nM Se/18 h., n = 10) and decrease of selenium plasma/selenium erythrocytes ratio from 0.939 to 0.791. Reduced glutathione (G SH) administration led to 1.5-fold decrease of plasma selenium level and 1.3-fold increase of selenium concentration in intestinal walls of sensitized rats (r = 0.720, P < 0.001). Chromatographic separation of plasma proteins showed that intragastric intubation of G-SH to sensibilized rats significantly decreased the protein P content and did not influence the concentration of Se-GSHPx, thus indicating the local selenium acceptor role of G-SH. G-SH administration did not influence the intestinal permeability in sensitised rats while use of complex additive: G-SH and selenium enriched spirulina--normalized the latter parameter and the ratio of protein P/Se-GSHPx in plasma. PMID- 10802883 TI - [The role on non-adrenergic regulation in the reaction of the rat pineal body to acute hypoxia and epithalamine administration]. AB - The influence of co-administration of alpha-methyl-p-tyrosine (inhibitor of catecholamine synthesis) and alpha-(+) beta-adrenergic antagonists under conditions of acute hypobaric hypoxia or pretreatment with epithalamin on cyclic nucleotide content in the pineal gland of juvenile male albino rats was investigated. Acute hypoxia was accompanied by increase of pineal level of cyclic nucleotides and administration of adrenoreceptor antagonists attenuated this effect. Pretreatment of animals with adrenoreceptor antagonists did not influence the effect of acute hypobaric hypoxia on pineal cyclic nucleotide content. This suggests involvement of non-adrenergic mechanisms into augmentation of pineal cyclic nucleotide level. Administration of epithalamin caused an increase of pineal cGMP. Administration of epithalamin to rats pretreated with adrenoreceptor antagonists increased pineal cGMP and to a lesser extent cAMP content. The latter suggests that epithalamin effect was not mediated via sympathetic innervation. It is concluded that non-adrenergic innervation and humoral regulatory mechanisms are obviously involved into activation of pineal gland under conditions of acute stress. PMID- 10802884 TI - [Pathogenetic basis for the use of GABA-ergic preparations in combination with atropine during chronic pain syndrome in patients with neurologic manifestations of lumbar osteochondrosis]. AB - The intensity of neuronal uptake of 3H- or 14C-labeled GABA, glycine, noradrenaline, choline and glucocorticoid receptor binding were investigated in different structures of the rabbit CNS by simulating chronic pain syndrome. Data obtained allow to suggest pathogenetically justified therapy including the complexes of phenybute + atropine and relanium + atropine. The contents of Leu Enkephaline, cortisol, insuline and insuline/cortizole ratio were studied in the blood serum of the patients with neurological manifestations of lumbar osteochondrosis and a long case history of the pain treated by the above mentioned complexes. The proposed therapeutical approach allowed to achieve pronounced antistress effect and regression of the pain syndrome due to stimulation of the CNS endogenic antinociceptive structures. It was shown that stress-produced hormones and opioids ratio in the blood serum may serveas indirect criteria of the functional activity of the endogenic antinociceptive structures before and after the treatment. PMID- 10802885 TI - [Biochemical consequences of acute and subacute tetrindole administration on rats]. AB - Acute and subacute administration of tetrindol resulted in pharmacological effects and biochemical changes which can be attributed to manifestations of toxic effect and activation of the detoxication systems. PMID- 10802886 TI - [Cytosol monoamine oxidase in the rat liver]. AB - Cytosolic and particulate monoamine oxidases have been isolated. Cytosolic preparation was free from mitochondrial and microsomal contaminations and also ribosome-bound MAO molecules. Cytosolic MAO had higher affinity for phenylethylamine and exhibited higher sensitivity to acetylenic inhibitors than the mitochondrial enzyme. PMID- 10802887 TI - [Effect of melatonin on wound healing and various biochemical characteristics of granulation-fibrous tissue in rats]. AB - The influence of melatonin application on wounds healing and biochemical composition of rat regenerating granulation tissue was studied. Melatonin decreased healing rate of wounds. The differences in electrophoretic pattern of proteins extracted by neutral saline solutions were detected. Melatonin increased quantity of neutral soluble collagen fraction and gene expression of minor types of collagen in normal skin. Spectrum of glycosaminoglycans' was changed, and earlier increase of chondroitinsulfats induced by administration of melatonin was observed. PMID- 10802888 TI - [Tissue-specific nucleoprotein complexes]. AB - A method of isolation of native nucleorprotein complexes from cattle cerebral cortex, thymus, and liver was developed. Compositions of these complexes were studied by means of gel-chromatography and ion-exchange chromatography. These preparations were shown to consist of several fractions of proteins and their complexes differ by molecular mass and electro-chemical properties. Native nucleoprotein complexes revealed high tissue specific activity, which was not species-specific. PMID- 10802889 TI - [Structural-functional changes in lymphocyte ans erythrocyte membranes after exposure to alternating magnetic field]. AB - The lymphocyte membrane fluidity, membrane potential, isoenzyme spectrum of superoxide dismutase (SOD) and catabolic product level of blood cell receptors in rats, healthy volunteers and patients with ovary and skin tumors were studied under the influence of low frequency electromagnetic field (EMF). The increase of fluidity of rat blood lymphocyte membranes was observed at 20 and 40 min exposure of the lymphocyte suspension to EMF. The increase of ANS fluorescence observed after 20 min exposure to EMF suggested a decrease of cell membrane potential. EMF exerted opposite changes of lymphocyte membrane fluidity of healthy volunteers and the effect depended on the initial level of this parameter. Study of SOD isoenzyme spectrum revealed that Cu,Zn-dependent SOD can account for the superoxide dismutase catalytic activity of supernatant and lymphocytes. The EMF exposure for 20 min caused 2-fold increase of SOD activity. The EMF exposure for 40 min caused the further increase of SOD activity but only in lymphocytes. At the same time the EMF exposure did not influence the actual SOD-activity in erythrocytes. However, there was clear anti-R-reagent-sensitive SOD-activity which was not abolished by DDC. The reasons and possible consequences of these changes of feuidity and SOD-activity under EMF effect are discussed. PMID- 10802890 TI - [Vitamin B 6 metabolism in phenylketonuria]. AB - Based on the analysis of the dependence of 4-pyridoxic acid urinary excretion from pyridoxal-5'-phosphate blood plasma level and its statistic distribution it has been shown that vitamin B-6 metabolism in children suffering from phenylketonuria and therefore the criteria of the body saturation with this vitamin differ from those for healthy people. Increased pyridoxal-5'-phosphate blood plasma level has been demonstrated for PKU children. The concentration of 11 ng/ml should be considered as a bottom border of the adequate supply with vitamin B-6. The elevated vitamin B-2 intake approximates vitamin B-6 status indexes of sick children to those usually measured in healthy children. The necessity for the reevaluation of vitamin B-2 and B-6 optimal diet content under this disease and its biochemical validation are discussed. PMID- 10802891 TI - [Hormone replacement therapy--women's expectations and fears]. AB - INTRODUCTION: To investigate attitudes, knowledge and fears concerning hormone replacement therapy (HRT) among women aged 50 to 70 years in Germany. MATERIAL AND METHODS: We conducted a representative survey of 1038 women aged 50 to 70 years including face-to-face interviews and written, comprehensive questionnaires in 1997. RESULTS: Current use of hormone replacement therapy (HRT) was 26%. Female gynecologists were more likely to prescribe HRT compared with their male counterparts. A majority of sampled women associated HRT with many benefits, the most significant of which is the improvement of climacteric symptoms. Specific knowledge was dependent on the level of education. Perceived negative side effects of HRT stated by two thirds of women were weight increase due to HRT, uterine bleedings (one quarter), and fear of breast cancer risk (17%). Approximately one third of all women objected against the use of HRT because this intervention would be "against nature". Major reasons to discontinue therapy within the first two years after initiation were either cessation of climacteric symptoms or negative side-effects. CONCLUSION: Women's expectations and fears related to HRT are primarily associated with the improvement of well-being and appearance by HRT. Prevention of osteoporosis and cardiovascular protection are of lesser importance in this context. PMID- 10802892 TI - [Effects of estrogens on the vascular wall: cellular and molecular mechanisms]. AB - There is evidence from animal models and in vitro experiments that estrogens exert direct effects on the vascular wall. 17 beta-estradiol mediates vasodilatation and inhibits smooth muscle cell proliferation/migration. Furthermore, it modulates the vascular inflammatory response by inhibiting cytokine activation and expression of cell adhesion molecules. Finally, 17 beta estradiol inhibits platelet aggregation and adhesion. Thus, vasoprotection by estrogens is mediated via genomic and nongenomic pathways. Estrogens regulate gene expression by interacting with estrogen receptors (ER) and other transcription factors. The possible estrogen actions include nongenomic stimulation of membrane mediators and second messenger systems. The existence of two ER subtypes alpha and beta indicates the potential of different biological estrogen effects. At present, the expression, the relation, and the biological actions of ER alpha and beta in different vascular beds of origin, in female and male as well as in healthy and atherosclerotic blood vessels are not known. PMID- 10802893 TI - [Secondary and tertiary prevention of atherosclerotic vessel disease]. AB - A first randomized, prospective, placebo-controlled, blind study--the so called HERS-Study--was designed to demonstrate that continuously combined application of 0.625 mg conjugated estrogens and 2.5 mg medroxyprogesterone acetate is a suitable treatment regimen in secondary and tertiary prevention of coronary artery atherosclerotic disease. After a 4.1 years duration of treatment no essential differences in the occurrence of reinfaction, other complications due to atherosclerotic alterations, and survival rates between verum and placebo groups could be evaluated. Unanswered ist, whether the duration of treatment was sufficiently long, the dose of estrogens high enough, or whether the mechanism of estrogen action is antagonized at least in part by continuously administered medoxyprogesterone acetate. Several other questions remain open due to scanty informations on LDL-concentrations, and a complete lack of data concerning insulin/insulin resistance, hyperglycemia, respectively. Better results may be obtained with sequential estrogen/progestin treatment regimes or with estrogens only, progestins added on demand. Remaining additional risk factors have to be treated. Life style and nutritional habits should be adapted to atherosclerosis. PMID- 10802894 TI - [Effect of hormone replacement therapy during the climacteric on cerebrovascular diseases]. AB - In epidemiological and animal studies it could be shown that a hormone replacement therapy decreased the risk of stroke. Sonographical blood flow measurement shows a positive effects to the cerebrovascular system caused by estrogen which was not modified by gestagens. In morphometric investigations by sonographical measurements of intima-media-thickness, it could be demonstrated that a positive influence by hormone replacement therapy can be achieved for a longer time. The intima-media-thickness, as a sign of a complex atherosclerosis, is decreased. The number of atherosclerotic plaques is significantly lower in patients receiving hormone replacement therapy (HRT) in comparison to a control group. A regression of the plaques by estrogen replacement therapy could be shown in one study. Although the group of patients suffering from stroke is heterogeneous, the benefit of HRT for prevention of cerebrovascular diseases could clearly be demonstrated. HRT is not contraindicated in patients suffering from cerebrovascular diseases. In most of cases, HRT is recommended in these patients. The therapeutic effect of HRT for cerebrovascular disease should be proved in further prospective studies. PMID- 10802895 TI - [Alternatives to hormone replacement therapy: raloxifene and tibolone]. AB - The bone is an active metabolic organ influenced by many substances like calcium, vitamin D, bisphosphonates etc. The postmenopausal osteoporosis is mainly caused by estrogen deficiency and hormone replacement therapy (HRT) has been shown to prevent the progress of osteoporosis. The following paper describes two alternatives to the classical HRT: raloxifen and tibolon. Raloxifen belongs to the selective estrogen receptor modulators (SERM) showing an estrogen-agonistic effect on bone. There is evidence that bone mineral density (BMD) is growing with treatment. In a three year study (MORE), a statistically significant decrease of lumbar spine fractures was demonstrated (RR 0.5-0.7). Furthermore there was a statistically significant reduction of receptor positive breast cancer (RR 0.10). Raloxifen shows beneficial effects on the lipids and does not induce endometrial proliferation. In the field of climacteric complaints, it is an estrogen antagonist and therefore inappropriate for this indication. Tibolon--a steroid hormone--and their three metabolites have estrogenic, gestagenic and weak androgenic effects on the different target organs. As expected, there is an increase of bone mineral density comparable to that of HRT or raloxifen; data of fracture rates with long-term therapy are missing. The substance and their metabolites are equivalent to HRT in the treatment of climacteric complaints. Tibolon shows some beneficial effects on the lipids and a lower bleeding rate compared to HRT. Raloxifen and tibolon are interesting alternatives to HRT which allow a more individual treatment of patients in the postmenopause. PMID- 10802896 TI - [Urinary incontinence--taboo during postmenopause]. AB - Urinary incontinence affects 30-60% of all women in the postmenopause. More and more women want to remove the taboo attached to incontinence, because they do not accept the reduction in quality of life, social and domestic activities. Detrusor instability (urge incontinence) and/or urethral sphincter incompetence (genuine stress incontinence) are the principal mechanisms, which lead to involuntary urine loss in older women. It is quite evident that additional factors as hypoestrogenism and urogenital atrophy influence urinary incontinence. Since in the postmenopause urge incontinence combined with urogenital atrophy its found predominantly, pharmacologic therapy with anticholinergic drugs and estrogen replacement are generally effective. Surgical therapy should be restricted to older women with genital prolapse and/or stress incontinence. In genuine stress incontinence, estrogen replacement therapy can lead to subjective improvement, but urodynamic parameters do not change objectively. PMID- 10802897 TI - [Dementia: risk and protective factors with special consideration of gender and hormone replacement therapy]. AB - This article focuses on the special role of women and their problems regarding the epidemiology and pathogenesis of dementia, especially Alzheimer's disease (AD). Recent epidemiological studies reveal higher rates of women affected by the two major dementing conditions, AD and vascular dementia (VaD). However, it is under debate whether a pure gender effect accounts for this, because gender differences e.g. in education, life expectancy, psychosocial development and health behaviour must be included in the analysis. Therefore it seems to be reasonable to have a look on gender related variances in other risk and protective factors of dementia. Recently, the menopause attracted the interest of dementia researchers, because retrospective studies revealed a favourable effect of hormone replacement therapy (HRT) on the incidence of AD by about 50%. However, in spite of well known "positive effects" of estrogens on the central nervous system, there is still not enough evidence for a recommendation of HRT for the prevention of AD and other dementias. PMID- 10802898 TI - [Sexuality of older women. The effect of menopause, other physical and social and partner related factors]. AB - Research on female sexuality in middle and old age (50-90 years) is reviewed. The descriptive results of the studies published up to now are summarized according to the dimensions of sexual activity (e.g. intercourse, tenderness, masturbation) and interest. The influence of various factors on female sexuality in middle and old age is critically discussed, namely health/body (health; general changes in sexual reactions; menopause), society (population structure; "double standard of aging"; historical-biographical influences) and couple relationship. Altogether, female sexuality is highly variable. The most prevalent psychosexual problems of older women are not the classic medical complaints (e.g. dyspareunia, insufficient lubrication) but a lack of tenderness and of sexual contact. The effect of the hormonal changes of the menopause is smaller than the effect of psychological, society- and partner-related factors. The so-called "Hormone replacement therapy" does not have significant effects on most sexual dimensions. PMID- 10802899 TI - [Breast cancer: assessment of individual risk and possibilities for prevention]. AB - Today, prevention of breast cancer (BC) is a great demand. The exact estimation of the individual BC-risk is a prerequisite for the participation on early cancer detection or the use of preventive medication or surgery. Various models for risk assessment of BC development or the presence of a predisposing mutation (i.e. BRCA1 or BRCA2) are used, but the statistical individual risk assessment still remains uncertain. Calculating an elevated risk or detection of a predisposing mutation leads to the recommendation of preventive measurements. After detailed assessment, prophylactic bilateral mastectomy is an option to consider for mutation carriers. For women with low BC-risk, chemoprevention can be discussed. Chemoprevention with tamoxifen (TAM)--indirectly supported by BC data from the raloxifen (RLX) prevention trial for osteoporosis and cardio-vascular disease- points to the right direction. Results from the three published TAM prevention trials are variable. Life time risk, age and life style have to be considered in the adapted individual risk-benefit assessment. The lack of long term risk data for chemoprevention and the effect on survival are arguments contra the routine use of TAM as a chemopreventive agent. PMID- 10802900 TI - [Practice guidelines--therapy of anxiety and compulsive disorders. Recommendations for therapy of anxiety and compulsive disorders of the Drug Commission of the German Medical Society]. PMID- 10802901 TI - Immortality. PMID- 10802902 TI - Small is still beautiful. PMID- 10802903 TI - Intentional dialog. PMID- 10802904 TI - More good intentions. PMID- 10802905 TI - Frontier medicine takes research strides. PMID- 10802906 TI - Where to with herbals? PMID- 10802907 TI - Psychotropic medications from natural products: a review of promising research and recommendations. AB - Most psychotropic agents employed in allopathic medicine have limited efficacy and significant side effects. Although usually beneficial, synthetic psychotropics are unavailable to approximately 80% of the world's population. Improved understanding of appropriate and safe uses of naturally occurring substances as psychotropic agents will greatly contribute to global mental healthcare. Empirical validation of non-allopathic treatments to ensure safety and efficacy is important because increasing numbers of patients in economically developed countries are using natural substances as medicinal agents. Patients and clinicians often lack accurate information, resulting in poor treatment outcomes or the possibility of drug-drug interactions when herbal medications are used with synthetic psychotropic medications. An important objective of this paper is to distinguish compelling scientific evidence supporting the use of natural products in psychiatry from political or institutional biases that have been misrepresented as scientific arguments. Following an overview of historical, legal, and regulatory issues, this paper presents findings of a systematic literature review on natural products used to treat neuropsychiatric disorders. Significant recent research is reviewed, including emerging treatments of seizure disorders, schizophrenia, dementia and age-related cognitive decline, depression, anxiety states, and substance abuse. Substantial evidence is advanced for safety and efficacy of many natural products used to treat neuropsychiatric symptoms or disorders. Preliminary findings suggest that several treatments based on natural substances are as effective and safe as synthetic pharmaceuticals in current use. Additional studies are indicated to confirm these findings, to elucidate mechanisms of action, and to elaborate standards for safe and appropriate treatment indications. In conclusion, strategic approaches aimed at facilitating improved networking, accelerating promising research directions, and enhancing quality standards of ongoing investigations into putative psychotropic agents from natural sources are recommended. PMID- 10802908 TI - Green tea monograph. PMID- 10802909 TI - Rational dosages of nutrients have a prolonged effect on learning disabilities. AB - CONTEXT: Reports that administration of nutrients has increased the academic performance of learning-disabled children exist in the literature. OBJECTIVE: To document the effects of nutrients on learning-disabled children in a controlled study. DESIGN: A randomized, double-blind, placebo-controlled crossover trial, which followed 1 year of open-label nutrients. Children who improved in the open label trial were eligible to enter the controlled phase of the study. SETTING: Subjects were enrolled from the general community through advertisements. PATIENTS OR OTHER PARTICIPANTS: Twenty children met the criteria for being learning disabled. INTERVENTION: Each child was tried out on some (but not necessarily all) of the B vitamins and minerals used in this study. These were administered semi-blinded for the first year; double-blinded in crossover rotations during the second year; and open-label in the ensuing years. MAIN OUTCOME MEASURES: At various time points, school-certified psychologists administered psychoeducational tests. School report cards were evaluated at baseline and for all subsequent periods. RESULTS: Twenty learning-disabled children entered the study, but 1 dropped out because of nausea. The remaining 19 children showed significant academic and behavioral improvements within a few weeks or months of open-label treatment with nutrient supplements. Some children gained 3 to 5 years in reading comprehension within the first year of treatment; and all children in special education classes became mainstreamed, and their grades rose significantly. Twelve of the children completed the 1-year double blind phase, after which approximately half of the children chose to remain on the nutrients for at least 2 additional years. For those who discontinued, it took at least 1 year to begin to see the first indications of decline in academic performance, and another year for their grades to drop significantly. In contrast, for children who remained on nutrients, the gains continued the upward trend; at the end of year 4, the difference in scores between the 2 groups had reached statistical significance (P < .01). CONCLUSION: The overall results of this study tentatively support the concept that learning disabilities may in some cases be a nutrient-responsive disorder. PMID- 10802910 TI - Tracy Gaudet the changing of medical education. Interview by Bonnie Horrigan. PMID- 10802911 TI - German clinic embraces holistic perspective. PMID- 10802912 TI - Damaging deliberations. PMID- 10802913 TI - Music therapy and melatonin. PMID- 10802914 TI - [Which language will MEDLINE speak in the next millennium?]. AB - OBJECTIVES: To analyze the trends in the prevalence of the languages used in Medline, since its creation up to the present time, for the overall indexed articles in this database and those specifically related to the field of Urology. The results are extrapolated to determine the trend over the next 15 years. METHODS: The Pubmed page on Internet (http:www.nlm.nih.gov/pubs/factsheets/nlm. html) and the Medline database (OldMedline 1960-1965 and current Medline 1966 1999) were accessed to determine the total number of indexed articles and the distribution by year, language and country. The same method was applied for articles specifically pertaining to the field of Urology. RESULTS: Overall, English accounts for 75% of the nearly 10 million articles indexed in Medline since its creation. The prevalence of the English language has grown steadily since 1966 when it already accounted for 53% of these indexed articles up to 89% in 1999, for a yearly increase of 1%. Similar results were found for the articles specifically pertaining to the field of Urology. The number of the indexed articles in the English language has grown to almost 400% since 1966, whereas those written in other languages have dropped by 40%. In some countries like Holland, Germany or Japan, most of the articles are published in English. CONCLUSIONS: If the current trend continues, all the articles indexed in the Medline database will comprise only those written in the English language by 2014. PMID- 10802915 TI - [Round cells in semen and genital infections]. AB - OBJECTIVE: The correlation between leukocytospermia and genital infections remains controversial. Similarly, it is inaccurate to consider the increase of round cells (RC) in semen as leukocytospermia. The purpose of this study was to determine the possible association between round cells, leukocystospermia and bacterial infections in infertile patients. METHODS: 410 semen samples were analyzed to determine sperm concentration, motility, morphology and round cells according to standard WHO citeria. Immature germ cells were differentiated by the Papanicolau stain, while the Nahoum-Cardozo method was used for polymorphonuclear leukocytes (PMN). Bacteriological studies in semen first urine sprout and urethral secretion were performed. Gram-Nicolle stain was utilized for the direct examination. Cultures were made in enriched blood Columbia agar and Thayer Martin agar was used for common germs. For mycoplasma identification, both A7 Sheppard solid and liquid media were used. Presence of Chlamydia trachomatis in urethral secretion was investigated by direct immunofluorescence. RESULTS: 79% (324) of the samples were dispermic, 9.02% presented more than 10(6) RC/ml. Of these, only 29.7% (11) presented leukocytospermia. Spearman's ranked correlation, used to measure association between RC/field and PMN leukocytes/ml, showed a value of 0.2705 with an associated probability of 0.1046, indicating a non-significant association between variables. Bacteriological studies performed in 32 samples showed 13 were positive, but only 4 (30.7%) presented leukocytospermia. Of the 19 samples with negative bacteriology, 15.78% were leukocytospermic. There was no statistically significant correlation between increased RC and the presence of germs (chi 2: 0.14965, p < 0.05) or between PMN and germs (chi 2: 1.01390, p < 0.05). CONCLUSIONS: These data suggest the convenience of performing bacteriological studies in semen in spite of the presence of RC in the direct examination. PMID- 10802916 TI - [Orthotopic substitutive cystoplasty in women. Technical modifications in anterior exenteration: results in 9 cases]. AB - OBJECTIVE: The results obtained in 9 female patients who underwent bladder substitution after a modified anterior pelvic exenteration are presented. METHODS: From 1994 to 1999, 9 women underwent bladder substitution after a modified anterior pelvic exenteration that preserved the two distal thirds of the urethra, the pubourethral ligaments and endo-pelvic fascia and limited laterovaginal dissection to preserve the innervation of the striated sphincter for continence. RESULTS: The bladder capacity was 332.9 +/- 35.6 ml and urinary flow was 17.7 +/- 2.9 ml/sec. Seven patients are completely continent and two patients have stress incontinence grade II. None of the patients had difficulty with bladder emptying. Fistula (one case) and diarrhea (one case) were the most important complications. CONCLUSIONS: Our approach is based on a better understanding of the anatomical mechanisms of female continence and the viability of the remaining urethra from the oncologic perspective. This technique achieves a high level of patient satisfaction and quality of life. PMID- 10802917 TI - [Renoureteral malformation and lithiasis. Therapeutic approach]. AB - OBJECTIVE: To analyze our results in the treatment of calculi in renoureteral malformations with extracorporeal shock wave lithotripsy (ESWL) alone and the efficacy of ESWL and adjuvant methods in the resolution of calculi in different types of renoureteral malformations. METHODS: The study comprised 141 cases of renoureteral malformations with urinary calculi; 68 were renal, 69 ureteral and 4 in the Bricker urinary diversion. All patients had been diagnosed by IVP and treated by ESWL. These patients were compared with a control group of 125 patients with calculi and a normal renoureteral system that were treated similarly. RESULTS: The malformations accounted for 2.61% of the 5,400 patients that had been treated in our unit from 1990-1998. The mean number of sessions for the patients with malformations was 1.5 versus 1.2 for the control group. The results of treatment were significantly lower for the group of patients with renal malformations than for the control group (54% vs. 87.2%, respectively). Similar results were obtained for the group of patients with ureteral malformations and the control group (87% vs. 75%). CONCLUSIONS: ESWL can be the first treatment option for calculi in patients with renal malformations, although they should be individually evaluated for status of the excretory system, urinary flow dynamics and size of the calculus in order to select the best treatment option. In patients with ureteral malformations, the same criteria utilized for those with a normal excretory system should be applied. PMID- 10802918 TI - [Clinical trial to evaluate trospium chloride (Uraplex) effectiveness and tolerance in patients with detrusor instability incontinence and its impact on quality of life]. AB - OBJECTIVE: To determine the efficacy, tolerance and quality-of-life effects of trospium chloride in women with overactive bladder. METHODS: An open, prospective multicenter trial was conducted on 75 women with urinary incontinence from overactive bladder (ICS criteria, urodynamic evaluation). Trospium chloride was administered at a dose of 20 mg twice daily for 8 weeks. Neurological examination and cystometry were performed at the start of the trial. Quality of life was evaluated by analogue visual scales (faces scale) and EUROQOL (health status scale). At the 4th week, urodynamic, clinical, quality-of-life and tolerance evaluations were performed. Clinical and tolerance data and quality of life index were assessed at the 8th week. RESULTS: 8 of the 75 patients did not complete the study. Thus, analysis of the therapeutic efficacy was performed in 67 patients, while description and tolerance analyses were performed for the overall group of patients. Urodynamic parameters significantly improved at 4 weeks: maximum bladder capacity (232.09 ml pre-treatment vs 315.83 ml post-treatment) and first desire to void (100.9 ml pre-treatment vs 156.7 ml post-treatment). Incontinence clinical items also improved. All quality-of-life indixes significantly increased at the 4 and 8 weeks control evaluation. Excellent or very good tolerance was observed in 89.5% of the patients. CONCLUSIONS: The results of the study corroborate the efficacy and tolerance of trospium chloride in the management of overactive bladder in women. Improvement in patient quality of life was also observed. PMID- 10802919 TI - [Urodynamics rationale. The flow curve]. AB - OBJECTIVE: To describe the mathematical representation of a common phenomenon: the urinary flow curve. METHODS: The statistical adjustment for minimum squares was the method employed in the study. RESULTS: A mathematical expression was obtained with a goodness of fit for each flow curve and an acceptable margin of error. CONCLUSIONS: The model which considers the flow curve as a parabola is not useful since this is an expression of a statistical phenomenon that varies from case to case. We utilized the minimum squares fit to obtain a regression curve that provides the nearest possible representation of urinary flow. Furthermore, we consider the flow curve to be the result of forces that develop in the detrusor, which produces the complex phenomenon of voiding. This study is the first step toward the development of a mathematical representation of the urinary flow mechanics. PMID- 10802920 TI - [Surgical treatment of hypospadias. Our experience and follow up in 1993-1998]. AB - OBJECTIVE: To review our series of hypospadias. The incidence of the different types of hypospadias and the complication rates according to the different surgical techniques and materials utilized are analyzed. METHODS: We reviewed our series of 130 patients with hypospadias who underwent surgical correction at the Department of Pediatric Urology from 1993-1998. RESULTS: The patients presented the following types of hypospadias: glandular (13 cases), balano-preputial (56), distal penile (42), mid-penile (9), proximal penile (2) and penoscrotal hypospadias (8). The surgical techniques utilized were the Mickulitz meatotomy procedure (6 cases), MAGPI (24), Mathieu (77), Crawford (5), onlay (7), Duckett (3), Retik (4), Duplay (1), Denis-Brown (2) and penoscrotal transposition (1). The complications commonly observed were fistula (20 cases), hematoma (4), stricture of urethral meatus (4), infection (3), megaurethra (3), skin necrosis (1). CONCLUSIONS: Surgical repair of this developmental anomaly is performed when the patient is approximately 18 months old. In our series 15% of the patients developed fistula, which is similar to the complication rate reported in the literature. However, since we started using monofilament resorbable material, the incidence of fistula has dropped from 20% to approximately 5%. Although other factors are involved, the lower incidence of fistula formation may be largely due to a reduced tissue reaction to foreign body. PMID- 10802921 TI - [Calcified bladder squamous carcinoma: radiologic considerations]. AB - OBJECTIVE: To report a case of squamous cell carcinoma of the bladder, an uncommon tumor type, that presented with wide calcified areas. The literature on calcified bladder tumors is reviewed with special reference to the diagnostic aspects. METHODS/RESULTS: The plain film, US and CT findings are presented. All the diagnostic imaging techniques demonstrated calcifications on the tumor surface and the characteristic curvilinear shape of this lesion. CONCLUSIONS: Calcified bladder carcinoma is rare and occurs in only 0.5% of the cases evaluated by conventional radiology. All calcifications probably arising in the bladder detected on conventional radiological evaluation should be studied further with other imaging techniques. If its etiology is unclear, cystoscopy with biopsy and/or a microbiological study should be performed. PMID- 10802922 TI - [Perirenal spontaneous hematoma in HIV(+)-patient in pelvic kidney]. AB - OBJECTIVE: A case of spontaneous perirenal hematoma in pelvic kidney in an HIV(+) patient is described. METHODS: The literature on the etiology of spontaneous perirenal hematoma in pelvic kidney is reviewed, with special reference to the three cases reported in HIV(+) patients. RESULTS/CONCLUSION: The patient presented none of the features described in the literature. The "watchful waiting" approach was adopted and the outcome was favourable. PMID- 10802923 TI - [Renal cell carcinoma associated with renal angiomyolipoma and renal cortical adenoma]. AB - OBJECTIVE: To report a case of renal cell carcinoma associated with renal angiomyolipoma and renal cortical adenoma and discuss the possibility of a hereditary etiology. METHODS: A 64-year-old patient with a tumor in the right kidney underwent right renal nephrectomy. The pathological findings revealed a clear cell renal carcinoma, two cortical angiomyolipoma and a renal cortical adenoma. The literature on these three coincident renal tumors is reviewed. RESULTS: The association of these three renal tumors is rare and is usually seen in patients with tuberous sclerosis. CONCLUSIONS: Renal angiomyolipoma is a benign tumor frequently associated with Bourneville's disease or tuberous sclerosis, a disorder of autosomal dominant transmission. Familial evidence of renal cell carcinoma has been previously reported, as in von Hippel-Lindau disease. It has been suggested that heredity may play an important role in the association of renal cell carcinoma and other renal tumors. PMID- 10802924 TI - [Bladder leiomyoma. Review of the literature and report of a new case]. AB - OBJECTIVE: To report an additional case of leiomyoma of the bladder and review the literature. METHODS: A case of leiomyoma of the bladder is presented. Its etiopathogenesis is described and the literature briefly reviewed, with special reference to the differential diagnosis. RESULTS/CONCLUSIONS: Leiomyoma of the bladder is an uncommon benign tumor that must be distinguished from other lesions. Treatment is by surgery, which generally achieves good results. PMID- 10802925 TI - [Retroperitoneal tumors. Retroperitoneal myxoid liposarcoma. Report of a new case]. AB - OBJECTIVE: To describe an additional case of a large retroperitoneal liposarcoma. METHODS/RESULTS: A case of a large retroperitoneal liposarcoma in a young male is presented. Treatment was by surgery. The clinical and pathological features, diagnosis, course and treatment of this lesion are discussed. CONCLUSIONS: Retroperitoneal tumors constitute a heterogeneous group of uncommon neoplasms of unknown etiology. Liposarcoma is the most frequent lesion in this group. It is usually asymptomatic and is often identified by the presence of a large abdominal mass. It is considered to be a peculiar lesion because of its biological and morphological features and course. PMID- 10802926 TI - [Hypernephroma associated with primary bladder tumor. Report of a case]. AB - OBJECTIVE: To present a case of tubular renal adenocarcinoma associated with transitional cell carcinoma of the bladder. METHODS/RESULTS: A case of tubular renal adenocarcinoma associated with transitional cell carcinoma of the bladder in a 42-year-old patient is described. Patient evaluation included intravenous urography, nephrotomograms, renal and bladder ultrasound evaluation and cystoscopic examination with biopsy. The patient underwent nephroureterectomy and partial cystectomy with subsequent radiotherapy and intravesical chemotherapy. At two-years follow-up, there is no evidence of disease progression. CONCLUSIONS: The appearance of various early-stage urological tumors in the same patient, is uncommon. Although a review of the literature showed that these cases were considered a medical curiosity, it should be taken into account that there is an increasing number of new cases. PMID- 10802927 TI - Treatment of retroperitoneal fibrosis with tamoxifen: case report and review of literature. AB - OBJECTIVE: To evaluate the efficacy of tamoxifen in the treatment of idiopathic retroperitoneal fibrosis in one patient and to review the results reported in the literature. METHODS: A 68-year-old man with idiopathic retroperitoneal fibrosis and obstructive acute renal failure was admitted to our department. Bilateral ureteral stents were placed and tamoxifen 20 mg daily p.o. was started. RESULTS: The ureteral stents were removed five months after tamoxifen therapy. IVP demonstrated normal appearance of the ureters nine months after medical treatment. An MRI scan showed an important decrease of the fibrotic periaortic mass at 12 months and then we stopped tamoxifen. CONCLUSIONS: Actually tamoxifen represents an attractive and safe choice of medical treatment for retroperitonea fibrosis, particularly in the acute stages. Nevertheless, the duration of treatment, the effectiveness and the persistence of the results are still uncertain because few cases have been reported in the literature. PMID- 10802928 TI - [Emotional parent-child bonding as a research topic]. AB - The author of the article introduces different themes of the study of emotional bonding between parents and children. Bonding behaviour is shown as significant factor influencing both psychical and physical health of children in different periods of their lives. Intergenerational emotional relationships create life background and the sense of stable, life certainty and safely. In this sense they become the source of active life of the individual in various ages. Several basic groups of factors, which participate on forming of these emotional bondings are further described in the text together with examples of the interventions making in the frame of family and health policy. Text represents the introduction to a series of articles which will open new knowledges about the formulation of bonding behaviour gained in the unique empiric study to a public. The research project "The Influence of the Family Life Style On the Course of the Pregnancy" was realised in the years 1996-1998 at the Research Institute of Child Health in Brno. PMID- 10802929 TI - [Nephrologic manifestations of mitochondrial diseases]. AB - Mitochondrial diseases, rated as rarities in paediatrics and neurology, become more and more often diagnosed in various body systems. In nephrology, mitochondrial defects play an important role in the pathogeny of tubular syndromes, interstitial nephritis, focal and segmental glomerulosclerosis, and probably also some other diseases. Review is aimed to point out such defects and to form conditions for their diagnosis and therapy. PMID- 10802930 TI - [Case-mix classification in post-acute and long-term care. Validation of Resource Utilization Groups III (RUG-III) in the Czech Republic]. AB - BACKGROUND: Czech facilities for non-acute, continuing care provide care for very heterogeneous group of residents with different clinical characteristics, care needs and resource use. The rate based funding of LTC doesn't reflect patient case-mix. Therefore, a case mix system RUG based on per diem resource use is being used abroad for rational and fair LTC funding. METHODS: The validity of RUG III has been evaluated and its use for financing of LTC and geriatric care in CR examined. In a sample of 1162 residents from 18 institutions patients were assigned to one of 44 RUG-III groups regarding their clinical characteristics. Data were analyzed using analysis of variance with individual care time per patient per day as independent and RUG-III groups as dependent variables. Weighted means for each group (case-mix indexes, CMI) were calculated. RESULTS: The RUG-III system achieved 59% variance explanation of total per diem costs of nursing and therapy/rehabilitation care and meets criteria of clinical validity. The CMIs for individual groups span from 0.39 to 2.70 i.e. differences in resource use between groups were sevenfold. Resource use within groups was relatively homogeneous. CONCLUSIONS: The RUG-III represent a suitable case-mix system for nonacute institutional care in the Czech health care. Besides its use for payment incentives, RUG-III can be used in facility management, quality assurance process and for comparative analyses on national and international level. PMID- 10802931 TI - [Inhalation of high-doses of ipratropium bromide (Atrovent) does not cause any changes in systemic or pulmonary hemodynamics]. AB - BACKGROUND: Parasympatolytics are recommended as the first choice remedy in patients with the chronic obstructive lung disease. Though their side effects are rare, some physicians are afraid of they can in high doses influence the cardiovascular system. The aim of the work was to ascertain whether the inhalation of 0.12 mg of ipratropium bromide might affect the systemic and pulmonary circulation. METHODS AND RESULTS: 15 patients (10 males, 5 females, average age 61.8 +/- 10.3 years) with chronic obstructive lung disease (13) and idiopathic pulmonary fibrosis (2) were examined during the period of disease stabilisation. Results of the right-side catheterization were not statistically changed (p < 0.91) 30 or 60 minutes after the inhaling of 0.12 of ipratropium. Also the value of the pick expiratory forced rate (PEFR) was not statistically increased (p < 0.25). CONCLUSIONS: Inhaling of 0.12 mg of ipratropium in patients with serious chronic pneumopathies did not affect systemic or pulmonary hemodynamics. Such dose appears to be safe even for patients with the chronic ischaemic hart disease. PMID- 10802932 TI - [How proteins seek their place. (Gunter Blobel, winner of the Nobel Prize for physiology and medicine in 1999]. PMID- 10802933 TI - [Bioethics in Asia]. PMID- 10802934 TI - Infectious diseases in England and Wales: July to September 1998. PMID- 10802935 TI - Mutation hotspots and DNA methylation. PMID- 10802936 TI - The role of DNA methylation in modulating Epstein-Barr virus gene expression. PMID- 10802937 TI - Promoter-region hypermethylation and gene silencing in human cancer. AB - In summary, it is apparent that alterations in DNA methylation are a fundamental molecular change associated with the neoplastic process and have important biologic implications for tumor initiation and progression. The promoter-region hypermethylation events covered in the present chapter are especially critical and can frequently serve as alternative mechanisms for coding-region mutations for loss of key gene function in neoplastic cells. The mechanisms underlying the precise role of this hypermethylation in gene silencing must be further defined, as must the determinants of the hypermethylation changes themselves. The therapeutic implications of promoter-region hypermethylation must be explored, and a potential use for establishing this change as a sensitive biomarker for use in multiple types of cancer-risk assessment and detection assays has already emerged. The next few years should see exciting advances in our understanding of an epigenetic process which, in conjunction with genetic alterations, appears to drive the process of neoplasia. PMID- 10802938 TI - Mammalian methyltransferases and methyl-CpG-binding domains: proteins involved in DNA methylation. PMID- 10802939 TI - Relationship between transcription and DNA methylation. PMID- 10802940 TI - DNA methylation, genomic imprinting and cancer. PMID- 10802941 TI - CpG-island methylation in aging and cancer. PMID- 10802942 TI - Mouse models in DNA-methylation research. PMID- 10802943 TI - DNA methylation inhibitors in the treatment of leukemias, myelodysplastic syndromes and hemoglobinopathies: clinical results and possible mechanisms of action. PMID- 10802944 TI - [Informational value of the prevalence of antiborrelia antibodies in a healthy and at risk population]. AB - The authors tried to confirm data on the prevalence of antiborrelia antibodies class IgM and IgG in the local healthy population (200 blood donors) and in subjects with an increased risk of infection (forestry labourers, forestry workers and huntsmen). All blood samples were analyzed in the National Reference Laboratory for Lyme borreliosis CEM (LB) in the National Institute of Public Health in Prague by the ELISA NRLB KC 90 method and in case of a positive result they were confirmed by the Western blot Lyme method. All patients completed a special questionnaire used as a standard procedure in the consultation centre for LB. In the group of blood donors 100 men were examined, mean age 33.5 years and 100 women, mean age 24.4 years. In 26 positive antibodies against Borrelia burgdorferi (Bb) were confirmed, but on invitation to medical examination only 22 attended, 11 men and 11 women (10.1%). Examination of antiborrelia antibodies by the ELISA method proved positivity of class IgM 7 times and of class IgG 9 times. By the Western blot method (WB) class IgG was confirmed three times. In the risk group 39 men were examined, mean age 40.6 years and 32 women. In 12 of them (17.9%) positivity of antiborrelia antibodies against Borrelia burgdorferi (Bb) was proved by the ELISA method as well as the WB method. If the percentage confidence interval of the control group is taken as a basis for comparison, then the value of the risk group is within this interval at a significance value of alpha = 0.01. In the conclusion the authors discuss the problem of interpretation of positive serological results and the pitfalls of their evaluation not only from the aspect of validity of laboratory findings, due to the absence of standardization of methods, but also with regards to obscure aspects of the persistence of IgM antibodies after years of lege artis antibiotic treatment. PMID- 10802945 TI - [Epidemiologic problems in hantavirus infections]. AB - The objective of the submitted paper was to present some information pertaining to epidemiological problems of hantavirus infection. In the Czech Republic the prevalence of these infections is not alarming but in the world cases of haemorrhagic fever with renal syndrome (HFRS) as well as hantavirus pulmonary syndrome (HPS) are reported. On the territory of the Czech Republic there are foci of hantaviruses and in conjunction with summer tourism or working activities abroad the import of this infection cannot be ruled out. With regard to the serious character of some forms of the disease it is essential to follow up this problem systematically. PMID- 10802946 TI - [Diagnosis and notification of rotavirus infection in the Czech Republic]. AB - In 1998 in 33 laboratories in the Czech Republic 1,223 rotavirus infections were revealed. The largest number of positive infections was recorded in children under the age of 3 years. The most frequent diagnostic procedure is latex agglutination which was used in 29 laboratories. In six departments the positive findings are reported directly to the appropriate epidemiological department. In the information system of the hygiene service, Epidat, in 1998 239 cases were reported, i.e. 19.5% of those confirmed by laboratory tests. The authors discuss the contemporary impact of rotavirus infections in the Czech Republic. PMID- 10802947 TI - [Seroprevalence of viral hepatitis A, B and C in intravenous drug users]. AB - Markers of viral hepatitis A-C of 99 intravenous drug users predominantly (in 96%) from the Karvina district were examined from March 1998 to February 1999. Pervitin (methamphetamine) was the basic drug in all subjects. Anti-HAV was detected in 6.4% of VHA non-vaccinated individuals, anti-HAV/IgM was not found in any case, HBsAg was detected in 4.0% (acute VHB was diagnosed in all these cases), anti-HBc in 9.2% and anti-HCV in 25.0% of the investigated cases. Our results confirmed the importance of drug use by injection, esp. needle sharing, for the spread VHB and VHC. In case of VHA the fecal-oral route of transmission in this subpopulation is dominant. PMID- 10802948 TI - [Health and economic benefits of compulsory regular vaccination in the Slovak Republic. I. Methods]. AB - The authors will submit in their series of contributions under the common title of "Health and Economic Benefits of Compulsory Regular Vaccination in the Slovak Republic" estimates of benefits and effectiveness of particular vaccinations. In their first contribution they deal with the objectives and methods of this evaluation essential for the allocation of funds for the maintenance of existing preventive programmes and also for the implementation of new preventive measures. On the basis of literary data and their own experience they formulate modified methods of cost/effectiveness and cost/benefit and other parameters adjusted for the conditions of the vaccination programme in the Slovak Republic. PMID- 10802949 TI - [Trichosporon--experience with species identification from human isolates]. AB - Among 162 human isolates of Trichosporon the author isolated according to "crucial" signs (Gueho et al. 1992, 1994) the following as dominant: T. mucoides (41%) and T. asahii (33%), T. inkin, T. ovoides and others as well as non pathogenic T. pullulans were rare in their investigation. To the originally most frequently reported type T. cutaneum belonged less than 10% of the total number of isolates. The variability of physiological and biological signs did not make classification of 5% isolated Trichosporon strains possible. The disc diffusion method confirmed resistance of the tested strains to 5-fluorocytosine, and inhibition by polyene and azole antimycotics. PMID- 10802950 TI - [Salmonelloses in the Czech Republic 1989-1998]. AB - In the Czech Republic in 1989 a triple increase of the incidence of salmonelloses was recorded: 34,435 cases. Since that year the morbidity trend varies between 400 and 500 cases per 100,000 population. The dominating agent (95% and more) is Salmonella Enteritidis PT8. The epidemic incidence was recorded on the whole territory of the Czech Republic mainly in Moravia and in the East Bohemian region. The specific morbidity is highest in 0 and 1-4 year-old children. The seasonal incidence has two peaks with the exception of 1997. During the period between 1989 and 1996 there was a significant increase of epidemics of salmonellosis in conjunction with food production incl. private confectionery shops, restaurants and the sale of foods in the streets. The most important vehicle are eggs and egg products, in particular confectionery. Salmonella Typhimurium DT104 is found only rarely in the Czech Republic. The first epidemic (15 cases) developed in 1998. PMID- 10802951 TI - Pain as a paradigm. PMID- 10802952 TI - The role of the new cell and molecular biology graduate program in medical education. PMID- 10802953 TI - A plan to improve pain management for dying patients. A proposal by the Governor's Blue Ribbon Panel on Living and Dying with Dignity Task Force on Pain Management in the Dying Patient, 1997. PMID- 10802954 TI - Pain management and palliative care in hospice. PMID- 10802956 TI - Invasive techniques utilized in chronic pain management. AB - A multidisciplinary approach to the treatment of chronic pain has been one of the most effective means of restoring the ability of a patient to return to an increased level of function. Many techniques used for diagnostic and therapeutic means have been utilized as part of this approach. This paper will describe some of the more common invasive techniques used in the treatment of chronic pain. PMID- 10802955 TI - Healing touch. PMID- 10802957 TI - To all who live and die in poverty or who suffer from observing it. An open letter on an ethic of humanity. PMID- 10802958 TI - Attitudes about care at the end of life among clinicians: a quick, reliable, and valid assessment instrument. AB - Several initiatives to improve care at the end of life involve educational programs to influence clinicians' attitudes about care for patients with terminal illnesses. The objective of this research was to develop and test a short and easily administered instrument for measuring physicians' and nurses' attitudes towards care at the end of life. The instrument was tested using a cross sectional study of 50 clinicians (25 physicians and 25 nurses) from general medicine, cardiology, oncology, and geriatric medicine. Both reliability and validity were assessed, and the instrument was found to have acceptable test retest reliability and construct validity. Such an assessment instrument may be useful in evaluating the impact of initiatives to modify attitudes towards terminal care and in improving the quality of care at the end of life. PMID- 10802959 TI - Comparison of CMA joint statement on resuscitative interventions and New Brunswick hospital corporations' policies on end-of-life treatments. AB - Why do most physicians have so much difficulty respecting the wishes of their terminally ill patients who refuse treatment? The normative pluralism model is introduced to answer this question. Comparative content analysis serves as the theoretical framework for evaluating the Canadian Medical Association Joint Statement on Resuscitative Interventions against the corresponding administrative policies of New Brunswick hospital corporations and relevant New Brunswick law. Despite protection afforded patients by law, fully 75% of New Brunswick hospital corporations' administrative policies permit physicians to ignore patients' expressed objection to treatments. The futility-of-treatment criteria in the CMA joint statement and in all provincial hospital corporations' policies authorize physicians to substitute their judgment for patients' expressed refusal of CPR. The author concludes that when medical professional norms conflict with the law, physicians tend to follow their professional normative order. PMID- 10802960 TI - Predictors of home death in palliative care cancer patients. AB - With recent changes in health care there is greater emphasis on providing care at home, including the support of families to enable more home deaths. Since a home death may not be practical or desirable in every family situation, there is a need for an objective way to assess the viability of a home death in each individual family situation. The purpose of this study was to describe the relative role of predictors of home death in a cohort of palliative care patients with advanced cancer. A questionnaire was created as a means of assessing the viability of a home death. Five questions were included. Ninety questionnaires were administered by home care coordinators. A follow-up questionnaire was administered to record the place of death. Of the 73 evaluable patients, 34 (47%) died at home and 39 (53%) died in hospital or hospice. The desire for a home death by both the patient and the caregiver, support of a family physician, and presence of more than one caregiver were all significantly associated with a home death. Logistic regression identified a desire for home death by both the patient and the caregiver as the main predictive factor for a home death. The presence of more than one caregiver was also predictive of home death. The questionnaire is simple and, if our results are confirmed, it can be used for predicting those who will not have a home death. PMID- 10802961 TI - Home versus hospice inpatient care: discharge characteristics of palliative care patients in an acute care hospital. AB - This prospective survey was initiated to identify factors that helped and hindered home discharge for 100 consecutive patients who did not require further specialist palliative or acute care. Information was collected on demographics, functional ability (using the Palliative Performance Scale [PPS] and Karnofsky Performance Scale [KPS]), cognitive function at discharge as measured by the Mini Mental State Examination (MMSE), home support circumstances, and patient and family preference for discharge. 59 patients were discharged home and 41 were transferred to a hospice. Younger patients with younger caregivers were discharged home more often. Patients with better MMSE and better functional ability (PPS and KPS) were also more likely to go home. Patients going home were more likely to be married. Preference for site of discharge was met for 76% of patients and 90% of families. Of the patients going to a hospice, 24% of patients and 7% of families preferred a home discharge. More physical support at home could have facilitated a home discharge for 13 patients. Functionally dependent and cognitively impaired patients were generally unable to return home. To support patients and their families in an environment of their choice, access to increased physical support in the home must be addressed. PMID- 10802962 TI - Music therapy in palliative care for hospitalized children and adolescents. PMID- 10802963 TI - Analysis of palliative care content in nursing textbooks. AB - Overall, this study demonstrates significant deficiencies in end-of-life care content in nursing textbooks. Defining palliative care, quality of life at the end of life, and issues of policy, ethics, and law are the foundation of end-of life care. Analysis of these topics revealed a need to clarify concepts and to apply them within the context of end-of-life care. The analysis also demonstrated a need to transfer findings from palliative care research and concepts from hospice into basic education. In March 1999 the investigators convened a conference in New York in collaboration with a group of medical investigators analyzing EOL content in medical textbooks. The conference was also attended by publishers, editors, and authors of textbooks. The investigators were very encouraged by their interest in and commitment to correcting any weaknesses in their books. The investigators provided them with specific recommendations for improvement and resources for locating both appropriate content and authors with palliative care expertise. These resources are also made available on the City of Hope Pain Resource Center Web site (http:@mayday.coh.org). Achieving the overall project goal of strengthened nursing education in end-of-life care will be reached through a variety of measures. Improving textbooks is but one important step. The various disciplines involved in palliative care can contribute to this project by working in collaboration with textbook publishers. PMID- 10802964 TI - A right to palliative care? What the United States Supreme Court did and did not say in the physician-assisted suicide cases. PMID- 10802965 TI - The no-CPR decision: the ideal and the reality. PMID- 10802966 TI - ADHD in adults. PMID- 10802967 TI - Munchausen by proxy. PMID- 10802968 TI - Citalopram for head-banging. PMID- 10802969 TI - Risperidone in Prader-Willi syndrome. PMID- 10802970 TI - Progesterone organogel for premenstrual dysphoric disorder. PMID- 10802971 TI - Tourette's disorder: epidemiology and comorbidity in primary school children. AB - OBJECTIVE: To study prevalence and comorbidity of Tourette's disorder in the general population of children and in a clinical setting. METHODS: School-age children in the general population and children attending a county-wide tic disorder clinic were screened and examined by the same doctor. Behavioral psychometric instruments with demonstrated reliability and validity were used. RESULTS: Depending on the sample characteristics, 0.15% to 1.1% of all children had Tourette's disorder. Boys outnumbered girls by 4:1 through 6:1. Attention deficits and empathy/autism spectrum problems (including Asperger's disorder) were very common, each type of comorbidity affecting approximately two thirds of individuals with Tourette's disorder. Overall behavior problem scores were high, and affected children exhibited a marked degree of functional impairment. CONCLUSIONS: Tourette's disorder is a common disorder with high rates of significant comorbidity. In most cases, attention deficits and empathy problems are likely to cause more suffering than the tics per se. PMID- 10802972 TI - Distinguishing illness severity from tic severity in children and adolescents with Tourette's disorder. AB - OBJECTIVE: To examine whether tic severity, comorbid disorders, or both are associated with illness morbidity in youths with Tourette's disorder (TD). METHOD: Subjects were 156 consecutively referred youths (aged 5-20 years) who met DSM-III-R criteria for Tourette's disorder at a major academic medical center. All subjects were evaluated with a clinical interview by a child and adolescent psychiatrist and an assessment battery that included the Schedule for Affective Disorders and Schizophrenia for School-Age Children-Epidemiologic version. Statistical analysis used chi 2 and multivariate logistic regression. RESULTS: Nineteen (12%) of the 156 youths with TD required psychiatric hospitalization. Current age, TD severity, TD duration, obsessive-compulsive disorder, psychosis, major depression, bipolar disorder, panic disorder, and overanxious disorder were significant univariate predictors of psychiatric hospitalization (p < .01). While tic severity was marginally significant as a predictor of psychiatric hospitalization (p < .05), major depression (p < .016) and bipolar disorder (p < .001) were robust predictors of psychiatric hospitalization, even after statistical adjustment for collinearity and correction for all other variables assessed. CONCLUSION: The findings indicate that comorbid mood disorders are strongly associated with illness morbidity in youths with TD, highlighting the importance of attention to comorbidity in patients with TD. PMID- 10802973 TI - Anxiety disorders and tic severity in juveniles with Tourette's disorder. AB - OBJECTIVE: It was hypothesized that comorbidity with anxiety disorders would predict tic severity in youths with Tourette's disorder (TD). METHOD: Subjects were 190 youths meeting DSM-III-R diagnostic criteria for TD who were consecutively referred to a pediatric psychopharmacology program between 1994 and 1997. Subjects were initially evaluated with a clinical interview and the Schedule for Affective Disorders and Schizophrenia for School-Age Children Epidemiologic version. Statistical analysis used t tests, chi 2 tests, and logistic regression analysis. RESULTS: One hundred thirty-four subjects were classified as mild/moderate and 56 as severe TD cases. No meaningful differences were found in socioeconomic status, gender distribution, or age at onset of TD between the 2 groups. The 2 groups could not be differentiated by the presence of comorbid mood or disruptive behavior disorders including attention-deficit hyperactivity disorder. Although obsessive-compulsive disorder (OCD) was overrepresented among the severe TD cases, the difference failed to reach statistical significance. Excluding social and simple phobias, all other anxiety disorders were more clearly overrepresented among subjects with severe TD; separation anxiety disorder most robustly predicted tic severity, irrespective of the presence of OCD or other anxiety disorders. CONCLUSION: Findings suggest that non-OCD anxiety disorders in general and separation anxiety disorder in particular may be significantly associated with tic severity in referred TD patients. PMID- 10802974 TI - Acute stress disorder symptomatology during hospitalization for pediatric injury. AB - OBJECTIVE: To examine and identify predictors of acute stress disorder (ASD) and ASD symptomatology (ASDS) in children hospitalized for injuries. METHOD: Fifty four youths were assessed while hospitalized for injuries. Dependent variables were parent and nurse ratings of children's ASDS. Independent variables included children's prior trauma exposure and behavior problems, injury severity and permanence, brain injury, injury or death to family/friend(s), parental distress, and child reports of the injury/hospitalization experience as meeting criterion A for ASD. RESULTS: A total of 92.6% of children felt the current experience met criterion A, compared with 64.8% of parents. According to parent questionnaires, 4 subjects (7.4%) met DSM-IV criteria for ASD while another 12 (22.2%) had clinically significant but subsyndromal ASDS. Children's ASDS, as reported by parents, correlated highly with parental distress and ratings of children's prior psychopathology, and modestly with injury severity and family/friend(s) injured or killed. Nurses' ratings of children's ASDS correlated strictly with injury- and accident-related variables, and not with parent ratings of children's ASDS. CONCLUSIONS: Children perceive injuries and hospitalizations as stressful. ASDS is widely though divergently reported by parents and nurses in children hospitalized for injury. Parental distress, children's prior psychopathology, and injury-related factors may be useful predictors of children's postinjury ASDS. PMID- 10802975 TI - Predicting posttraumatic stress after hospitalization for pediatric injury. AB - OBJECTIVE: To determine the prevalence and predictors of posttraumatic stress disorder (PTSD) in children after hospitalizations for accidental injuries. METHOD: Forty-eight children (aged 7-17 years) and their parents were assessed during hospitalization with measures of children's prior traumatization, prior psychopathology, injury severity, parental acute distress, and child acute distress. At outpatient follow-up at least 1 month later, children were evaluated for current PTSD diagnosis and PTSD symptomatology (PTSDS) by a child structured interview and for PTSDS by a parent questionnaire. RESULTS: A total of 12.5% had the full syndrome of PTSD at follow-up, and an additional 16.7% had partial (sub syndromal) PTSD. Full PTSD was associated with a higher level of prior psychopathology, higher parental acute distress, and higher rates of prior sexual abuse, compared with partial or no PTSD. Prior psychopathology, parental distress, and, to a lesser extent, children's acute distress as reported by parents and breadth of prior traumatization, predicted subsequent PTSDS. CONCLUSIONS: Full or partial PTSD is relatively common in youths 1 month or more after hospitalization for injuries. Parents' acute distress as well as children's prior psychopathology, prior traumatization, and acute distress may be useful predictors of such injured children's subsequent PTSD or PTSDS. PMID- 10802976 TI - Parent's perceptions of their first graders' need for mental health and educational services. AB - OBJECTIVES: (1) To estimate the proportion of first graders perceived by their parents as needing mental health or educational services, (2) to examine the validity of these perceptions, and (3) to examine whether factors found to be associated with children's referral to or use of services are associated with parental perceptions. METHOD: Subjects are an epidemiological sample of 467 first graders. Children's social adaptational status and psychological well-being were assessed by multiple methods. Logistic regression was used to investigate the associations between indicators of need and parental perceptions of their children's need for services. RESULTS: More than 39% of the children were perceived by their parents as needing services. Predictors of parents perceiving their children as needing services were parent and teacher ratings of children's behavior and symptoms, children's standardized achievement test scores, and gender. CONCLUSIONS: Given that a large proportion of parents reported their children as needing mental health and educational services and that parents play a central role in children's receiving services, longitudinal studies that investigate the entire process by which children enter into and are maintained in the service delivery system are needed. PMID- 10802977 TI - Mothers' mental illness and child behavior problems: cause-effect association or observation bias? AB - OBJECTIVE: A number of studies have consistently found that a mother's mental health (particularly her level of depression) is a strong predictor of mental health problems experienced by her child(ren). However, the validity of this finding is in doubt because the majority of these studies have relied on maternal reports as indicators of children's behavior. METHOD: This prospective, longitudinal study examines data on the mental health of the mother from prior to the birth of her child to when the child reaches 14 years of age. Child behavior is measured at 14 years of age using reports from mother and child. Mother and child responses are compared to provide an indication of the possible magnitude of maternal observation bias in the reporting of child behavior problems. RESULTS: Anxious and/or depressed mothers tend to report more cases of child behavior problems than do their mentally healthy counterparts or children themselves. Differences between mothers and youths in reporting behavior problems appear to be related to the mothers' mental health. CONCLUSIONS: Current maternal mental health impairment appears to have a substantial effect on the reporting of child behavior problems by the mother, thereby raising questions about the validity of reports of child behavior by persons who are currently emotionally distressed. PMID- 10802978 TI - Predictors of adolescent suicide attempts: a nationally representative longitudinal study of Norwegian adolescents. AB - OBJECTIVE: To investigate the risk and protective factors for previous and future suicide attempts among adolescents. METHOD: A representative sample of high school students (N = 9,679) in grades 7 through 12 (aged 12-20 years) were followed from 1992 to 1994. Response rate was 97% at initial testing and 80% at follow-up. Measures of psychiatric symptoms (depressed mood, eating problems, conduct problems), substance use, self-worth, pubertal timing, social network, and social integration were included. RESULTS: A total of 8.2% had ever attempted suicide and 2.7% reported an attempt during the 2-year study period. Logistic regression analysis showed that future attempts were predicted by previous attempt, female gender, young age, perceived early pubertal development (stronger among girls), suicidal ideation, alcohol intoxication, not living with both parents, and poor self-worth. CONCLUSIONS: The importance that the clinician ask about previous suicidal behaviors is underscored. Early pubertal timing (particularly among girls), loss of self-worth, and alcohol intoxication may serve as risk factors for future suicide attempts. PMID- 10802979 TI - Aggressive behavior in clinically depressed adolescents. AB - OBJECTIVE: To investigate the prevalence and characteristics of aggressive behavior in adolescent inpatients and outpatients with major depressive disorder (MDD). Differences between males and females in prevalence and type of aggression, and level of parent-child agreement in report of aggression, were analyzed. METHOD: Participants were 74 adolescents with MDD, aged 13 to 17 years. The Structured Clinical Interview for DSM-IV Axis I Disorders was used to identify MDD. Adolescents' aggressive behavior was assessed using an adapted version of the Brown-Goodwin Assessment for Lifetime History of Aggression; the Measure of Aggression, Violence, and Rage in Children; and the Buss-Durkee Hostility Inventory-Adapted Version. RESULTS: Results indicate high levels of aggressive behavior in adolescents with MDD. Amount and type of aggression did not differ by gender. Results indicate poor correspondence between parent and adolescent reports of aggression, which was most marked for females. CONCLUSIONS: Aggressive behaviors are highly prevalent in depressed youths, with similar types and levels evident in males and females. Parents tend to under-report and may not be cognizant of aggressive behavior that occurs outside the home, particularly for females. PMID- 10802980 TI - A double-blind, placebo-controlled study of Adderall and methylphenidate in the treatment of attention-deficit/hyperactivity disorder. AB - OBJECTIVE: While Adderall has been available for the treatment of attention deficit/hyperactivity disorder (ADHD) for several years, there are few controlled studies comparing it to methylphenidate. METHOD: Fifty-eight children with ADHD (mean age 8.1 +/- 1.4 years) were randomly assigned to receive placebo, methylphenidate, or Adderall in a double-blind, parallel-group design for 3 weeks. Dosage was adjusted at the end of weeks 1 and 2 via an algorithm based on teacher and parent ratings. Final doses were 12.5 +/- 4.1 mg/day for Adderall and 25.2 +/- 13.1 mg/day for methylphenidate. Teacher and parent ratings, as well as the psychiatrist's Clinical Global Impression (CGI), were the final outcome measures at the end of week 3. RESULTS: Both medications were superior to placebo at reducing inattentive and oppositional symptoms in the classroom and on the CGI. Adderall produced significantly more improvements on teacher ratings and the CGI than methylphenidate, although the algorithm may have limited dosing in the methylphenidate group. Seventy percent of children in the Adderall group were given medication once a day, compared with 15% of the subjects receiving methylphenidate. CONCLUSIONS: Adderall compared favorably to methylphenidate, and the behavioral effects of Adderall appear to persist longer than those of methylphenidate after individual doses. PMID- 10802981 TI - Driving outcomes of young people with attentional difficulties in adolescence. AB - OBJECTIVE: To examine associations between attentional difficulties at age 13 and a range of adverse driving outcomes measured at 21 years. METHOD: Data were gathered over the course of a 21-year longitudinal study of a birth cohort of 1,265 New Zealand children. Data collection included the following: (1) parent and teacher report measures of attentional difficulties (13 years); (2) measures of driving behavior, including involvement in an accident, drinking and driving, and traffic violations (18-21 years); and (3) measures of a range of potentially confounding individual, sociofamilial, and driving-related factors. RESULTS: Young people with high levels of attentional difficulties were at greater risk of involvement in a motor vehicle accident, drinking and driving, and traffic violations. These associations were largely explained by the personal characteristics (gender, conduct problems) and driving experience (length of time respondent held a license, distance driven) of young people with attentional difficulties. Even after adjustment for the effects of confounding factors, adolescent attentional difficulties placed young people at increased risk of an injury accident, driving without a license, and other traffic violations. CONCLUSIONS: Associations between adolescent attentional difficulties and subsequent driving risks largely reflect the effects of confounding factors correlated with attentional difficulties and driving outcomes. However, even after adjustment for confounding, adolescent attentional difficulties contributed to later injury accident risk and possibly also to risky driving behavior. PMID- 10802982 TI - Adrenomedullary function during cognitive testing in attention deficit/hyperactivity disorder. AB - OBJECTIVE: Reported correlations between epinephrine (EPI) excretion and classroom performance, the cognition-enhancing effects of EPI infusion, increased EPI excretion with stimulants, and reports of decreased EPI excretion in attention-deficit/hyperactivity disorder (ADHD) suggest that sympathoadrenomedullary function might be altered in ADHD. This hypothesis was tested by examining sympathetic and adrenomedullary functioning during cognitive testing in boys with diagnosed ADHD. METHOD: Urinary excretion of EPI and norepinephrine during a 3-hour cognitive test battery was assessed in 7- to 13 year-old boys. Excretion rates (nanograms per hour per square meter of body surface area) were determined in 200 individuals with ADHD (diagnosed according to DSM-IV criteria), with or without co-occurring oppositional defiant/conduct disorder or learning disorder. A non-ADHD contrast group (n = 51) with similar comorbidity was also studied. RESULTS: Substantially lower (mean +/- SE) urinary EPI excretion was observed in the ADHD-inattentive subtype (n = 71) compared with the control group (200 +/- 22 versus 278 +/- 24 ng/hr/m2; F = 5.99, p = .015, critical alpha = .017). No diagnostic group differences were seen for norepinephrine excretion. Correlational analysis of both parent- and teacher rated behaviors revealed that inattention factors consistently negatively predicted urinary EPI excretion. CONCLUSIONS: The data extend findings of lower adrenomedullary activity during cognitive challenge in individuals with ADHD and suggest that the alteration is associated with inattentive behavior. PMID- 10802983 TI - Executive function oculomotor tasks in girls with ADHD. AB - OBJECTIVE: To assess executive function in girls with attention deficit/hyperactivity disorder (ADHD) using oculomotor tasks as possible trait markers for neurobiological studies. METHOD: Thirty-two girls aged 6 to 13 years with DSM-IV ADHD and 20 age-matched, normal control girls were tested on a variety of oculomotor tasks requiring attention, working memory, and response inhibition, which included smooth pursuit, delayed response, and go-no go tasks. RESULTS: Girls with ADHD performed the delayed response task correctly on 32% of trials as measured by number of memory-guided saccades, in contrast to 62% of trials for control subjects (p = .0009). Patients made twice as many commission errors to no go stimuli (p = .0001) and 3 times as many intrusion errors (saccades in the absence of go or no go stimuli; p = .004) during the go-no go task compared with controls. Smooth pursuit performance was statistically equivalent across subject groups. Repeated testing in a subgroup of 15 patients revealed substantial practice effects on go-no go performance. CONCLUSIONS: The data confirm that girls with ADHD exhibit impairments in executive function, as has been reported in boys, implying a similar pathophysiology of ADHD in both sexes. However, practice effects may limit the utility of the oculomotor go-no go task for some neurobiological studies. PMID- 10802984 TI - Visual sustained attention in a child psychiatric population. AB - OBJECTIVE: To increase knowledge of the diversity and specificity of sustained attention deficits in children with attention-deficit hyperactivity disorder (ADHD), with special reference to the issue of distinguishing between children with ADHD and children with other psychiatric diagnoses. METHOD: A visual sustained attention task was used to compare 52 boys with ADHD with 55 normal controls, 29 boys with oppositional defiant disorder or conduct disorder (ODD/CD), 29 boys with anxiety or dysthymia (ANX/DYS), 43 boys with pervasive developmental disorder, 24 boys with ADHD plus ODD/CD, and 14 boys with ADHD plus ANX/DYS. RESULTS: Compared with normal controls, children with ADHD were slower, were more inaccurate, were more impulsive, were less responsive to feedback, and showed less perceptual sensitivity and stability of performance, resulting in a marked decrease in vigilance over time. Unresponsiveness to feedback and the extent of the decrease in vigilance during time on task were found to be the only factors that distinguished children with ADHD from children with other diagnoses. CONCLUSION: Although only children with ADHD are characterized primarily by "attention deficit," sustained attention deficit is common to a certain extent to all children with psychiatric disorders. PMID- 10802985 TI - Developing child abuse prevention, identification, and treatment systems in Eastern Europe. AB - Although the dissolution of the Soviet Union in 1989 brought a welcome end to the Cold War, the subsequent reestablishment of independent Eastern European countries revealed a host of problems. Among these was frequent child abuse and neglect, although lack of epidemiological data precluded any accurate estimate of prevalence. In an effort to address the problem of child abuse within this rapidly changing sociopolitical environment, the Children's Mental Health Alliance and the Soros Foundation organized the Eastern European Child Abuse and Child Mental Health Project in 1995. The authors describe the development of this multidisciplinary initiative involving collaboration with professionals from 17 countries in Eastern Europe. They highlight the goals, principles, and major phases of the project, as well as some of the challenges involved in its implementation. PMID- 10802986 TI - Factitious disorder by proxy in which the presenting problem is behavioral or psychiatric. PMID- 10802988 TI - Y2K.3. Hippocrates redux PMID- 10802987 TI - Genetics of childhood disorders: XIV. A gene for Rett syndrome: news flash. PMID- 10802989 TI - Seventy-five years ago... PMID- 10802990 TI - Call for diazepam replacement. PMID- 10802991 TI - Gender-based personality traits and military cohesion. PMID- 10802992 TI - Knowledge attainment, perceptions, and professionalism in participants completing the didactic phase of an Army reserve critical care nursing residency program. AB - Combat hospitals in today's Army demand nurses with critical care nursing "8A" additional skills identifiers. The intensity of future wars and operations other than war, together with highly technological weapons, forecast a large number of casualties evacuated rapidly from combat with wounds that require skillful and intensive nursing care. Many of the critical care nurses providing future care are positioned in the reserve components and require creative approaches to education and training concentrated into one weekend per month. An Army Reserve critical care nursing residency program was designed in one midwestern combat support hospital. The didactic course, phase I, was evaluated for effectiveness in achieving outcomes of increased knowledge attainment, enhanced perceptions of critical care nursing, and higher degrees of professionalism. Twenty-seven registered nurses completed the course, and 30 nurses from the same hospital served as controls. A repeated-measures analysis examined outcomes before intervention (time 1), at course completion (time 2), and at a 6-month follow-up (time 3). The course was effective at increasing scores on knowledge attainment and perceptions of critical care nursing; however; professionalism scores were initially high and remained so throughout the study. This research extends information about critical care nursing education and evaluates a training mechanism for meeting the unique requirements and time constraints of nurses in the reserve components who need to provide a high level of skill to soldiers in combat. PMID- 10802993 TI - The effect of oral pyridostigmine bromide nerve agent prophylaxis on return of twitch height in persons receiving succinylcholine. AB - Prophylactic pyridostigmine is used for troops under threat of organophosphate exposure and can affect the neuromuscular blocking agent succinylcholine. The purpose of this study was to determine what effect pyridostigmine would have on neuromuscular blockade from succinylcholine. Thirty-nine subjects were enrolled in this double-blind, placebo-controlled investigation and assigned to either the pyridostigmine or the placebo group. All subjects were evaluated for pyridostigmine or placebo side effects and given an anesthesia induction that included the neuromuscular blocking agent succinylcholine. Neuromuscular blockade was measured by twitch height with a nerve stimulator. Interval data were analyzed using the chi 2 test and Student's t test. The pyridostigmine group demonstrated a prolonged recovery time to return of 25% (p = 0.003) and 75% (p = 0.028) of twitch height compared with the placebo group. No differences were noted for any other parameter measured. We conclude that the recovery from neuromuscular blockade after succinylcholine is prolonged while the intubating conditions remain unaffected. PMID- 10802994 TI - Army psychiatry in the Korean War: the experience of 1 Commonwealth Division. AB - This study seeks to investigate the incidence of psychiatric casualties in 1 Commonwealth Division during the Korean War. It had been hypothesized that these casualties were unusually low compared with earlier conflicts. Casualty returns and psychiatric reports were analyzed and showed that the war fell into two phases determined by the intensity of combat, which, in turn, influenced the nature of the psychiatric disorders encountered. Rates of acute combat stress were closely correlated with battle casualties, although not with total psychiatric admissions or nonbattle injuries. The limitations imposed on the psychiatric liaison service by the medical organization suggested that the incidence of psychosomatic cases, including cold injury, may have been unintentionally underreported. PMID- 10802995 TI - Confidentiality and the psychological treatment of U.S. Army aircrew members. AB - The present article addresses the issue of confidentiality in U.S. Army psychological services and the special considerations affecting the confidentiality afforded to Army aviation personnel receiving such services. The author reviews Army regulations and American Psychological Association ethical standards relevant to the issue of confidentiality for aircrew members. Recommendations are offered for mental health professionals who provide services to Army aviation personnel, and a hypothetical clinical case is presented to illustrate the concepts discussed. PMID- 10802996 TI - Quadricepsplasty after war fractures. AB - Knee movements after fractures caused by explosive devices, as well as after intra-articular fractures of the knee, are often inadequate. This paper presents the results of quadriceps-plasty performed in 10 patients with the purpose of improving knee function. All of the patients were treated by the external fixation method, either after femoral fractures caused by explosive devices or for intra-articular knee fractures. All of them manifested markedly decreased knee flexion (15-70 degrees, with an average of 32 degrees). After quadricepsplasty and physical therapy, the achieved knee flexion was enough for normal walking (80-130 degrees, average 97.5 degrees). Mean knee mobility was increased 65.5 degrees. Our paper presents indications, methods, results, and complications for quadricepsplasty performed after war injuries. PMID- 10802997 TI - Attention to detail: injuries at altitude among U.S. Army Military static line parachutists. AB - Altitude injuries, defined as injuries sustained by military static line parachutists before ground impact, have not been reviewed for 50 years. There are indications that these injuries are increasing at Fort Bragg, North Carolina. Between May 1, 1994, and April 30, 1996, surveillance was conducted for parachute related injuries and altitude injuries at Fort Bragg. The incidence of all jump related injuries was 8.1/1,000 aircraft exits. Significant risk factors for injury included being 30 years of age or older (p < 0.001) and being female (p = 0.003). The overall incidence of altitude injuries was noted to be 0.46/1,000 aircraft exits. Risk factors for altitude injury included being 40 years of age or older (p = 0.005) and in the rank of E-1 to E-3 (p = 0.0001). Fifty-four percent of injuries occurred during exiting before complete parachute deployment, and 46% occurred during the opening shock of the parachute. Mechanisms of injury included static line entanglement (33%), riser/suspension-line entanglement (46%), aircraft strikes (21%), unsecured equipment strikes (1%), and opening shock deceleration (1%). The majority of severe altitude injuries are caused by riser/suspension-line entanglement (63%) and involve the knee joint (37%). Although the incidence of altitude injury is quite small, the potential risk for career-threatening and/or life-threatening injury is great. This risk can be reduced by appropriate training and attention to detail. PMID- 10802998 TI - The effect of full protective gear on intubation performance by hospital medical personnel. AB - OBJECTIVE: To assess the influence of protective gear on intubation performance. DESIGN: Prospective, controlled measurement of duration and quality of intubations performed on mannequins by medical personnel with and without protective gear in a crossover design. PARTICIPANTS: Eight teams each comprising an anesthesiologist and a nurse. RESULTS: Intubation duration with and without chemical warfare gear was 69.2 +/- 7 and 47.3 +/- 6 seconds (mean +/- SEM), respectively (p < 0.05). Moreover, rating of intubation quality as "very good" by the anesthesiologists declined significantly from 62.5% without chemical warfare protective gear to 6.25% with the garment and mask. Tube fixation was the rate limiting step when performed with protective gear (p < 0.05); it was assessed by 81% of the anesthesiologists as the critical step. A learning curve was not observed during the study. CONCLUSION: Protective gear causes a significant prolongation of intubation duration; however, endotracheal intubation can be performed effectively. Technical improvements are warranted for tube fixation because it is the critical step. PMID- 10802999 TI - Radiation injuries from military and accidental explosions: a brief historical review. AB - Nuclear radiation was discovered in the late 1800s. Advances in nuclear physics split the atom to herald the atomic age soon afterward. Now, a century later, health care providers remain acutely aware that nuclear hazards may present instantly and unexpectedly, with devastating and massive results. This article highlights known radiation injuries from military and accidental explosions. This information is critical for the preparedness of health care providers in view of aging nuclear power plants, potential industrial and medical accidents, and the buildup of military weapons by other countries. PMID- 10803000 TI - The comparative radiopacity of Fuji IX-GP, an intermediate restorative material. AB - The radiopacity of intermediate restorative materials should be sufficient to enable the clinician to distinguish the material from normal and decalcified tooth structure. The purpose of this study was to determine the relative radiopacities of intermediate restorative materials, including a newly introduced high-viscosity, self-cured, condensable glass ionomer material. Radiographs were made of six intermediate restorative materials: two reinforced zinc oxide-eugenol materials (IRM and Zinroc), a conventional glass ionomer material (Ketac-fil), a synthetic resin material (Cavit), a eugenol-free zinc oxide material (Tempit), and a new, general-purpose, condensable glass ionomer material (Fuji IX-GP). Optical density was measured using a densitometer. The optical density of dentin and enamel were used for radiographic comparison. Statistical analysis revealed significant differences among materials: Cavit = IRM = Tempit > Zinroc = Fuji IX GP > Ketac-fil = enamel > dentin (where > indicates a statistical difference at p < or = 0.05). Although not as radiopaque as some other intermediate materials tested, the radiopacity of Fuji IX-GP appears sufficient to aid diagnosis. PMID- 10803001 TI - Pediatric diseases and operational deployments. AB - Many nations now export military health as a proactive arm of the nation's contribution to the maintenance of international peace in trouble regions of the world; and all nations are called upon from time to time in emergency and disaster situations to help out in their regions of interest. Children and young teenagers constitute some 50% of war-stricken populations. This paper explores this increasingly important role of military medicine from the point of view of a practicing pediatrician and career doctor-soldier. Many international operational deployments undertaken in the last 5 years have required the insertion of pediatric clinical and preventive health resources. Deployments to Rwanda, the countries of the former Yugoslavia, Somalia, Bougainville (in Papua New Guinea), Irian Jaya (in Indonesia), and the Aitape tsunami disaster response (the Sepik region of Papua New Guinea) have all necessitated major pediatric interventions. In some operational deployments, in excess of one-third of patient and clinical contacts have involved the care of children, including clinical treatments ranging from life-saving resuscitation to the care of children with both tropical and subtropical illnesses. They have also involved mass immunization campaigns (e.g., in Rwanda) to prevent measles and meningococcal septicemia. In developing countries, at any time approximately 1 to 4 teenage and adult women is pregnant; and of these, 1 in 15 is suffering a miscarriage during any 2-week period. The implications of this audit are that service members must be multi-skilled not only in the traditional aspects of military medicine and nursing but also in (a) the developmental aspects of childhood; (b) the prevention of infectious childhood diseases by immunization and other means; (c) the recognition and management of diseases of childhood; and (d) the management of the normal neonate and infant, especially those orphaned in refugee disaster and other emergency situations. Doctor-soldiers hold special credentials to be advocates for the protection of children caught up in armed conflict and its aftermath. In this context, advocacy to ban anti-personnel land mines is topical, because unfortunately the more than 2,000 deaths and injuries each month involve children and their families, often injured long after the cessation of hostilities. Pediatric issues are now the business of all who serve in the health disciplines in the broader profession of arms. PMID- 10803002 TI - Adverse reactions to smallpox vaccine: the Israel Defense Force experience, 1991 to 1996. A comparison with previous surveys. AB - The aim of the present study was to assess the post-smallpox vaccination complication rate in a cohort of Israel Defense Force recruits enlisted in the calendar years 1991 to 1996 and to compare it with rates reported, in similar age groups, in large surveys during the 1960s. The overall complication rate was 0.4 per 10,000 vaccinees, and the rate of severe complications was very low, similar to previously published data. We conclude that among young healthy adults, vaccination with smallpox vaccine is relatively safe and is associated with a low rate of complications. Severe complications were very rare in this age group in our study. However, the complication rate is increasing with the increased percentage of primary vaccinees. PMID- 10803003 TI - Providing quality medical care at a remote, isolated duty station. AB - The U.S. armed forces deploy active duty personnel to virtually every region of the world. Family members accompany the service member at many of these remote, isolated assignments. This discussion highlights the barriers to the provision of medical care to these active duty personnel, their families, and other eligible beneficiaries, such as government service and Department of Defense civilians. To succeed in this endeavor, the isolated military medical treatment facility must first consider the critical resources available that will enable it to overcome known barriers. Next, careful deliberation will identify the necessary components of a health care network suitable for the remote duty station. The facility must then recognize and address its responsibility to conduct ongoing evaluation of the quality of care and of customer satisfaction with its network. This discussion concludes with a description of a successfully implemented health care network at a remote, isolated duty station. PMID- 10803004 TI - Characteristics of hormone-treated prostate carcinoma: stressing the need for clinician-pathologist communication. AB - OBJECTIVE: Patients scheduled for prostatectomy often receive androgen deprivation therapy to make malignant tumors more amenable to resection and improve the postoperative course. These hormonal therapies may significantly alter the histomorphology of carcinoma of the prostate detected on subsequent needle biopsies. METHODS: Needle specimens were obtained from resected prostates harboring biopsy-proven carcinoma previously treated with leuprolide. The tissue was examined by light microscopy to note architectural and cytologic characteristics. RESULTS: A high proportion of treated carcinomas had an atrophic, infiltrative appearance. Nuclear and nucleolar enlargement were consistently observed. Macronucleoli, blue-tinged mucin, and intraluminal pink amorphous material was frequently identified. CONCLUSIONS: The markedly atrophic nature of the cells and glands may result in either overgrading of prostate carcinoma or failure to recognize the more subtle patterns of this malignant neoplasm. It is imperative that clinicians convey a history of hormone treatment to pathologists when core biopsies of the prostate are submitted for histologic evaluation. PMID- 10803005 TI - Complications from cardiac catheterization: analysis of a military database. AB - BACKGROUND: Cardiac catheterization is a common procedure in the United States. Our purpose was to assess possible risk factors for complications from cardiac catheterization. METHODS: The Civilian External Peer Review Program database, which contains data on 3,494 cardiac catheterizations performed at 28 military facilities from 1987 to 1989, provided the patient population for this study. Of 360 abstracted clinical elements, 27 were selected by a panel of internists and cardiologists for evaluation as potential risk factors and were analyzed using logistic regression. Complications were analyzed within three categories: major (myocardial infarction, cerebral vascular accident, or death within 24 hours of catheterization); minor (hemorrhage requiring transfusion, pseudoaneurysm, fistula, or femoral thrombosis); and any. RESULTS: The mean age of the 3,494 patients was 56 years, and 75% of them were male; 85% were white, 10% were African-American, and 5% were other races. Complication rates were as follows: death (N = 13), 3.7/1,000; cerebral vascular accident (N = 16), 4.1/1,000; myocardial infarction (N = 22), 5.6/1,000; hemorrhage (N = 20), 5.1/1,000; fistula (N = 7), 0.3/1,000; and thrombosis (N = 15), 3.8/1,000. These were categorized as 59 major, 71 minor, or 122 any complications. Complications were more likely in patients with hypertension (odds ratio, 1.8; 95% confidence interval, 1.05-3.18), peripheral vascular disease (odds ratio, 2.9; 95% confidence interval, 1.1-8.7), age greater than 60 years (odds ratio, 2.1; 95% confidence interval, 1.2-3.8), and those undergoing angioplasty (odds ratio, 6.0; 95% confidence interval, 2.9-12.2). CONCLUSIONS: Hypertension, age greater than 60 years, peripheral vascular disease, and procedures either nonelective or involving angioplasty all independently increased the risk of complications. There was a "dose-response" relationship between risk and number of risk factors. The risk of a complication may be greater than 10% in patients with more than three risk factors. PMID- 10803006 TI - Solid waste management practices in U.S. Army medical treatment facilities. AB - U.S. Army medical treatment facilities (MTFs) will be challenged to reduce waste as the Army internalizes as environmental ethic and privatizes solid waste management. A questionnaire was modified to survey solid waste management practices, participation in waste reduction and recycling programs, and solid waste management problems in 25 MTFs. Questionnaires were returned by 19 (76%) of the sites. Eighteen sites participated in waste reduction and recycling programs. Twelve sites used contractors to disinfect potentially infectious solid waste off site. Mean importance ratings of waste reduction and recycling were 2.00 and 1.83 (1 = very important; 5 = very unimportant), respectively. Limited staffing was ranked as the most significant waste management problem, followed by cost and regulatory compliance. More information on waste generation in MTFs is needed. The Army Medical Department should be presented as a model of environmental and natural resource stewardship for the health care industry. PMID- 10803007 TI - A model for demand management in a managed care environment. AB - Downsizing of personnel and budgets demands effective means of managing patients' demand for care to ensure that care assets are appropriately utilized. Preliminary data indicate that the Central Triage concept of demand management is effective, efficient, and satisfactory to patients. As a centralized portal to care for individuals presenting without appointments, registered nurses screen and triage via protocol. The nurses send patients to the Emergency Center, make same-day appointments, or provide self-care education as indicated. During the first 9 months of operation, Central Triage inprocessed 35,231 patients. Twenty three percent were determined to have self-care deficits and received education to assist them in meeting their current self-care demands. Follow-up of 10% of the patients receiving self-care education revealed that 88% improved without further intervention and 95% were satisfied with the care they received. Central Triage has demonstrated an annual cost avoidance exceeding $2,500,000. PMID- 10803008 TI - The cost-effectiveness of varicella screening and vaccination in U.S. Army recruits. AB - Varicella outbreaks in the U.S. Army disrupt training, reduce readiness, and represent substantial costs. Vaccination of susceptible individuals may be cost effective. We conducted a cost-effectiveness analysis comparing screening of all incoming recruits and vaccination of susceptible individuals at either initial entry training (IET) or medical entrance processing station (MEPS), universal vaccination at IET, and no intervention. Primary health outcomes included the number of varicella cases prevented during the 8-week initial training period. The varicella hospitalization rate was 21.6 per 10,000 per year. In 100,000 recruits, 36 cases of varicella are expected at a cost of $181,000 in the absence of an intervention. Screening at IET would prevent 4 cases but would cost an additional $3,255,000 more than no intervention. Screening at MEPS would prevent 3 cases and save $521,000 per case prevented during the IET but would cost $2,734,000 more than no intervention. Universal vaccination would prevent 2 cases but would cost $15,858,000 more than MEPS screening and $18,592,000 more than no intervention. These results are robust. Cost per case of varicella prevented ranged from $390,000 to $7.9 million. Scarce prevention resources could be more cost-effectively allocated to other prevention programs. PMID- 10803009 TI - Doxycycline-induced esophageal ulceration in the U.S. Military service. AB - U.S. military forces are frequently deployed with little warning to regions of the world where chloroquine-resistant malaria is endemic. Doxycycline is often used for malaria chemoprophylaxis in these environments. The use of doxycycline can be complicated by esophageal injury. Two cases of esophageal ulceration will be discussed, followed by a review of the literature. Doxycycline causes esophageal injury through a combination of drug-specific factors, the circumstances of drug administration, and individual patient conditions. Patients with dysphagia attributable to esophageal ulceration are managed by intravenous fluid support and control of gastric acid reflux until their symptoms resolve over 5 to 7 days. The risk of esophageal injury can be minimized by use of fresh capsules, drug administration in the upright position well before lying down to sleep, and drinking at least 100 ml of water after swallowing the medication. PMID- 10803010 TI - Tactical management of urban warfare casualties in special operations. PMID- 10803011 TI - [The Decade of the Bones and Joints: 2000-2010]. PMID- 10803012 TI - [Pathogenesis of porphyria cutanea tarda]. AB - Porphyria cutanea tarda (PCT) results basically from decreased activity of uroporphyrinogen decarboxylase (UROD) in the liver. PCT is not a homogeneous disease; it can be either inherited or acquired. Not only alterations at the UROD gen locus but also other genetic factors outside the locus take part in the inactivation of UROD, that support polygenic inheritance of PCT. In every case, acquired factors take also part in development of the overt form of PCT. Iron has a key-role in the oxidative damages. PMID- 10803013 TI - [Prevalence of chronic viral hepatitis in drug abusers]. AB - Authors examined the prevalence of hepatitis B, C and D viral infections in Hungarian drug users. Between January 1995 and October 1998 256 examinations were made (58% intravenous, 42% non intravenous drug user). Hepatitis C virus infection in 27 patients, hepatitis C and B virus infection in 4 patients, hepatitis B virus infection in 17 patients was detected. Every hepatitis B virus positive case was past infection. Hepatitis D virus infection was not detected. Clinically overt liver disease was proved in more than half of the hepatitis C virus infected patients. Because of insufficient collaboration only 11 were followed up. Liver biopsy was made in 5 cases. Interferon therapy was indicated in 3 cases. The 24% of intravenous drug users was anti-HCV seropositive contrary to 1.4% of non intravenous group. Anti-HCV seropositivity was proved in 38% in common needle users, while in disposable needle users only 3%. The prevalence of hepatitis C virus infection in intravenous drug users is rather frequent in Hungary too. The exact diagnosis of liver diseases is very difficult as for insufficient collaboration. The prevalence of hepatitis B virus infection in i.v. and non i.v. drug users is the same as in the normal population. The importance of information, especially to avoid common needle use is stressed. PMID- 10803014 TI - [Pre- and perinatal aspects of hemophilia A and B]. AB - Authors investigate in a retrospective study obstetrical and genetical data in 20 years period of 149 pregnancies of patients turning to genetical counselling because of haemophilia A and B. In case of heterozygote mother there have been fetal determination of sex, and in case of male fetus, there have been DNA examination in 23 of the 35 cases. In case of sick male fetus the couple made a decision on keeping the pregnancy or not, knowing well the genetical risk. Haemophilia A occurred in case of 135 pregnancies (98 pregnancies of 55 heterozygote mothers, and 37 pregnancies from 20 sick fathers). Haemophilia B occurred in case of 14 pregnancies (9 pregnancies of 3 heterozygote mothers, and 5 pregnancies from 4 sick fathers). In case of haemophilia A heterozygote pregnant women there were 32 proven male fetuses, and in 22 cases there have been DNA examinations. In 16 cases there have been artificial abortions (in 10 cases proven disease by DNA examination), and 4 sick male newborns were born from the 16 deliveries (the disease was proven during pregnancy by DNA examination). One male newborn (healthy) was born from the 3 proven male fetuses of haemophilia B heterozygote pregnant women, in 2 cases there have been artificial abortions (in one case on the basis of DNA diagnostics). In cases of heterozygote mothers (haemophilia A and B altogether) the ration of the spontaneous abortions was 13.1%. The rations of the premature deliveries (8.2%) and the Caesarean sections (8.2%) were not higher than the national average. The ration of the bleeding complications during pregnancy was 18.7%, in 2.7% of the cases transfusion was necessary. In case of sickness of the father (in heterozygote female fetuses the haemostasis may change from the fetal side) the ration of the bleeding complications during pregnancy was 18.2%. In connection with delivery, obstetrical bleeding complications occurred in 12.2%, atonia in 2%, abrasion after delivery in 4.1, transfusion in 10.2% in cases both of haemophilia A and B heterozygote mothers. From the neonatological complications in one case there was cerebral haemorrhage, and in one case bleeding from the umbilical stump. (Both newborns were male with haemophilia.) In connection with delivery there was no haematoma developing on the skull of the newborns, there was no need of giving transfusion. In case of sickness of the fathers the ration of the instrumental uterine examination was 6.7%, there were no neonatological and other obstetrical complications. PMID- 10803016 TI - [About departmental libraries or thoughts in connection with the memorial meeting "50 years of Medical Libraries"]. PMID- 10803015 TI - [Castleman's disease]. AB - Castleman's disease is an example of the so-called atypical lymphoproliferative disorders. The optimal therapy of this morphologically and clinically heterogeneous disease is largely unknown. The authors report three cases of multicentric Castleman's disease (two hyalin vascular and one mixed variant). They analyze the pathogenesis, clinicopathologic features and management of this rare entity and review the literature. PMID- 10803017 TI - [75th anniversary of Dr. Friedrich Trendelenburg's death. (Data, photographs, documents)]. PMID- 10803018 TI - [Experiments of cough. 1875]. PMID- 10803019 TI - [The beginnings of electrocardiography in Hungary]. PMID- 10803020 TI - [The pathogenesis of juvenile bronchial asthma]. PMID- 10803021 TI - [REP and ERIC repetitive DNA sequences in bacteria--diagnostic significance]. AB - Main part of eukaryotic genomes is build of unique sequences coding proteins and RNAs, but they contain as well numerous repeats interspersed with single-copy fragments. Existence of repetitive sequences were also demonstrated in prokaryotic genomes. They are found in different species of Gram-negative and Gram-positive bacteria. Interspersed repetitive sequence elements called REP and ERIC sequences are present in different species of Enterobacteriaceae family, including Escherichia coli and Salmonella typhimurium. Their functions are not completely clear, probably they play important role in regulation of gene expression. Nevertheless, REP and ERIC elements are widely use in identification and genetic analysis of bacteria. For example, using rep-PCR technique it is possible to discriminate between closely related serovars of the same species, which enables to analyze phylogenetic and epidemiological relations among them. PMID- 10803022 TI - [Chemical modifications of macromolecules and their use in preparation of protein antitumor drug conjugates,vaccines and diagnostic reagents]. AB - This article reviews high temperature glycation of proteins, and describes some applications of thus received neoglycoconjugates as vaccines or drug and gene carriers. The use of macromolecules: antibodies, fibrinogen and natural or synthetic polymers as antitumor drug carriers is discussed. Moreover the article presents method for monitoring of proteins modification by glutaraldehyde. PMID- 10803023 TI - [Antitumor activity of gamma delta T lymphocytes]. AB - Increasing knowledge about the role played by gamma delta T lymphocytes in antitumor immunity is of importance to both the fundamental and clinical sciences. gamma delta T cells are also characterized with reference to theirself neoplastic transformation and development of leukemias or lymphomas with gamma delta fenotype. PMID- 10803024 TI - [Immunity in Toxoplasma gondii infections]. AB - The article presents selected data concerning pathogenesis, clinical manifestations, natural and specific resistance and vaccines against T. gondii infections. Toxoplasma gondii protozoan has been recognized as one of the most successful parasites infecting any nucleated cell of human individuals and most warm-blooded animals. The infection causes life-threatening disease in individuals with defective immunity such as fetuses, AIDS patients and transplant recipients. In immunocompetent humans toxoplasmosis is usually asymptomatic. Following an acute phase of infection characterized by systemic spread of rapidly dividing tachyzoites, bradyzoites encyst in various host tissues (brain, muscles) and may persist there lifelong. Toxoplasmosis in controlled by vigorous cell mediated immune response capable of killing infected cells and parasites. As shown in the mouse experimental model natural killer cells (NK) are critical for the resistance at the early stage of primary infections, whereas adaptive immunity depends on T lymphocytes. Both CD4+ Th1 and CD8+ Tc1 lymphocytes are believed to mediate protection by producing of IFN-gamma, a pitoval cytokine that induces anti-Toxoplasma effector mechanisms in macrophages. PMID- 10803025 TI - [Experimental hepatocarcinogenesis--evaluation of the significance of theoretical models. Contemporary models]. AB - There are an increasing number of models for the study of liver cancer development with such agents as chemicals, hormones, and viruses. This process is almost always a multistep and during the long period of cancer development, discrete cells or cell populations acquire step-by step the various properties that go to make up a cancer. The hepatocarcinogenesis models are useful in identification and analysis of the preneoplastic and neoplastic alterations, as well HCC. This article presents a review of theoretical foundation and mechanisms of various models of experimental hepatocarcinogenesis as well contain a short presentation of animals practical in those experiments, principally rats. The remarkable similarities between many models with different carcinogens in animals and humans suggest the importance of such studies in understanding of molecular basis of liver cancer development. PMID- 10803026 TI - [Structural elements and conformation of basement membranes]. AB - This article presents the latest information about biological functions of basement membranes. It describes all important basement membranes' components and their structure. Schemes showing modular sequence of collagens, nidogen, laminins and other polypeptides and also structural model of basement membranes were included. PMID- 10803027 TI - [Mutations of genes coding collagen type I--biochemical and clinical effects]. AB - Type I collagen is the most abundant structural protein of human body. In this paper the effects of mutations in COL1A1 and COL1A2 genes on biochemical properties of this protein and clinical manifestations are described. PMID- 10803028 TI - [Best wishes 2000]. PMID- 10803029 TI - [Image of the month]. PMID- 10803030 TI - [Pharma-clinics. How I treat ... hypo- or hyperkalemia]. AB - The treatment of dyskalemia must be primarily etiological and later symptomatic. When moderate but significant hypokaliemia exists (K < 3 mmol/l or 3.5 mmol/l with cardiac disease), oral K supplements must be/given. The intravenous route must be reserved for emergency or impossible oral administration. Acute and severe hyperkalemia (K > 7 mmol/l) must be first corrected by different intravenous measures with secondary oral intervention. In chronic and/or moderate hyperkaliemia (K > 5.5 mmol/l), the oral route for treatment is preferred. In any case, the research of the cause (mainly drug induced) is fundamental. PMID- 10803031 TI - [Clinical case of the month. Case report of adrenal metastases from lung adenocarcinoma]. AB - A 29 years old woman was diagnosed and lobectomized for a left upper lobe adenocarcinoma classified T1N0M0. Three months later a PET Scan revealed a lesion in the left adrenal which was then surgically removed and confirmed metastatic. Polychemotherapy and radiotherapy of the surgical site were started. Unfortunately four months later, a control PET Scan followed by biopsy showed a right adrenal metastasis which grew rapidly in site of the treatment. The patient died one year later in dissemination. We suggest that PET Scan should be performed before lung neoplasm resection. Unique and synchronous adrenal lesions should be removed because they do not respond any therapy. If these metastatic locations appear shortly after lung resection, according to this case and the literature, it seems that surgical resection is not useful. PMID- 10803032 TI - [Antioxidant vitamins in the prevention of cardiovascular diseases. First part: epidemiologic studies]. AB - The hypothesis of the atherogenic role of oxidized. LDL lipoproteins and the observation of the longevity of individuals on a "mediterranean diet" led to the concept that antioxidant vitamins may exert cardiovascular protective effects. In this first article, we summarize the results of the main epidemiological studies which analyzed the influence of dietary intakes (or resulting plasma concentrations) in vitamin E (alpha-tocopherol), vitamin A (beta-carotene) or vitamin C (ascorbic acid). The three types of studies available in the literature provided quite heterogeneous results, in favour of a protective role of antioxidant vitamins as far as cross-cultural and most prospective observational studies were concerned, but rather negative when considering case-control studies. However, in general, epidemiological studies tended to support, a beneficial role of these antioxidant vitamins to prevent cardiovascular diseases, at least in some subgroups of individuals. PMID- 10803033 TI - [Influenza: from vaccine prevention to antiviral therapy]. AB - Influenza is a highly infectious disease responsible for dangerous epidemics, especially in patients at high risk. The vaccine exerts a valuable protective effect estimated up to 70% and is still considered as the key-approach against influenza. Antiviral agents of the first generation (amantadine, rimantadine, ribavirine) have limited use because of poor tolerance and occurrence of resistance. Zanamivir or Relenza, marketed by Glaxo Wellcome, is a new virostatic drug acting as a neuraminidase inhibitor. It prevents the release of new viruses and so stop the propagation of the infection. It must be taken orally by inhalation within 48 hours after the onset of symptoms. The treatment lasts 5 days (10 mg twice daily). Its efficacy has been demonstrated in controlled clinical trials, and its tolerance is generally excellent. However, caution is recommended in patients with asthma and chronic bronchitis because of the potential risk of bronchoconstriction. No resistance has been detected until now. Zanamivir is active on all strains of influenza A and B viruses. PMID- 10803034 TI - [Tea time ... a privileged moment in preventing cancer?]. AB - Several recent epidemiological and biological studies suggest that polyphenols contained in tea exhibit anticarcinogenic effects. The present review is a synthesis of the current information. PMID- 10803035 TI - [Childhood in danger]. AB - Child abuse is a human production resulting from the failure of three essential characteristics of the human condition: the possibility of attachment, speech and the capacity to love. It is perpetuated though trans-generational transmission which traps the individual in an obvious incapacity to occupy a people position, inducing confusion of the roles and statute of the place of each in the family or in the society but more especially in the interpersonal relation. The child abuse must be seen by the professional as an obvious incapacity of the victims but also of their abuser, who are often them-selves former abused children, to develop a relation based on the respect and the love of others. The therapy is first of all an accompaniment of one and the other towards the differentiation and the possibility of installation of stable and reliable constructive representations. In short, an opening towards the expression of the denial of suffering and the rebuilding of a new ideology based on love and creativity. PMID- 10803036 TI - [Place of the Ross procedure (pulmonary autograft) in aortic valve surgery]. AB - The pulmonary autograft operation consists of an aortic replacement using the autologous pulmonary valve. The pulmonary valve is substituted by a cryopreserved pulmonary homograft. This operation is in fact a delicate double valve replacement whose benefits are linked to the viability of the new aortic substitute. The pulmonary autograft has superb hemodynamic features and very low thrombogenicity. The report of a selected observation offers the occasion of defining the current indications of this operation in aortic valve surgery. PMID- 10803037 TI - [Diagnosis of primary hyperparathyroidism: role of scintigraphy]. AB - We report a case of hyperparathyroidism whose diagnosis was suspected by a clinical history and confirmed by routine laboratory tests. Scintigraphy has permitted to localize a focalized lesion corresponding to an adenoma endocrinopathy. The new developments of this imaging technique will be discussed further. Some aspects of physiopathology will be briefly considered and a therapeutic algorithm will be proposed. PMID- 10803038 TI - [Amazing epic of blood transfusion...]. AB - On the occasion of the 100th anniversary of the discovery of blood groups by Karl Landsteiner, the author makes a historical note on the amazing history of the blood transfusion from the origin to the beginning of the XXth century. PMID- 10803039 TI - [How I investigate ... thoracic artery dissection]. AB - Thoracic aortic dissection constitutes an acute pathology that requires a rapid, accurate and reliable diagnosis. The authors describe and illustrate the advantages and limitations of imaging modalities used in this pathology. PMID- 10803040 TI - [Pharma-clinics. Medication of the month. Telmisartan (Micardis)]. AB - Telmisartan is a new non peptide angiotensin II receptor antagonist which selectively and insurmountably inhibits the angiotensin II AT1 receptor subtype without affecting other receptor systems involved in cardiovascular regulation. In the treatment of essential hypertension, it shows a dose-dependent activity within the dose range of 20 to 80 mg per day. The maximum effect is obtained by 80 mg per day and it is very close to that seen with larger dosages. Telmisartan has an extremely long half life of 20 to 30 hours and its efficacy is maintained throughout the 24 hour period after one single daily intake. Telmisartan is excreted almost exclusively in the feces (99%). Side-effects are comparable to placebo. PMID- 10803041 TI - [Pharma-clinics. Medication of the month. Moxonidine (Moxon)]. AB - Moxonidine (Moxon, Solvay Pharma) is the first member of a new class of centrally acting antihypertensive agents. The selective activation of central I1 imidazoline receptors results in an inhibition of peripheral sympathetic activity and produces arterial vasodilatation. Moxonidine is indicated in the treatment of essential arterial hypertension, at a usual daily dose of 0.4 mg (initial dose of 0.2 mg/day), in one administration per day. Its tolerance profile is better than that of other centrally-acting antihypertensive agents which stimulate alpha-2 adrenergic receptors, and similar to that of medications of other classes. As monotherapy, the antihypertensive efficacy of moxonidine is similar to that of most other antihypertensive agents. In case of insufficient response, moxonidine may be associated with other antihypertensive compounds, which leads to a better efficacy with no deterioration of its good tolerance profile. PMID- 10803042 TI - [Clinical study of the month. The STOP-2 study of arterial hypertension in the elderly]. AB - After the demonstration of the efficacy of beta-blockers or diuretics versus placebo to prevent cardiovascular complications in elderly hypertensive patients in the first STOP-Hypertension study in 1991, a Swedish group published at the end of 1999 the STOP-2 Hypertension study. The latter randomised trial showed in a similar population that the cardiovascular protection of more recent antihypertensive agents such as calcium antagonists and angiotensin-converting enzyme inhibitors is similar to that of the conventional antihypertensive drugs used in the first study. In fact, the degree of blood pressure control appears to be more important than the type of antihypertensive drugs used, and this conclusion is reinforced by the observation that numerous patients should rapidly be treated by more than one antihypertensive agent to reach blood pressure targets. PMID- 10803043 TI - [Thoracic CT scan in intensive care unit patients: information supplementary to thoracic radiography]. AB - PURPOSE: To assess the diagnostic value of thoracic CT-scans in comparison with conventional chest radiographs in ICU-patients. METHODS: Chest radiographs and corresponding thoracic CT-scans of 25 consecutive surgical ICU-patients were reviewed and interpreted independently by two radiologists. We analyzed the additional information provided by CT-scans and the diagnostic and therapeutic relevance of these findings. RESULTS: In 22 patients (88%), thoracic CT revealed 35 single additional findings, in comparison with the corresponding bedside radiographs. In 7 cases (28%), these findings (tube-malpositioning, mediastinitis, mediastinal abscess and pneumothorax) required modification of patient management. Of 7 patients with tube thoracostomy, 3 tubes were malpositioned, which was only depicted on CT-scans. In 10 of 21 cases (48%), pleural effusions could only be visualized by CT. 3 of 5 (60%) pneumothoraces were detected by CT-scans only. There were no significant complications during transport or CT-examination. CONCLUSION: Thoracic CT may provide significant information in addition to plain chest radiographs, particularly when specific problems or questions, referring to the management of ICU-patients, arise. PMID- 10803044 TI - [Renal parenchymal defects as signs of renal tuberculosis]. AB - Findings of urogenital tuberculosis in computed tomography have been reported seldom although this manifestation of the tuberculous disease is on the second place following pulmonary tuberculosis. We report on a 52 year old women suffering by culturally proven urogenital tuberculosis. The enhanced computed tomography showed hypodense lesions clearly on the cortex border of the kidney. Other findings of renal tuberculosis reported in the current literature as hydrocalices or hydronephrosis were not seen at all. We postulate that the described finding is characteristic for an early stage of renal tuberculosis. PMID- 10803045 TI - [Use of patient-related contrast media protocols in cardiovascular computerized tomography]. AB - PURPOSE: To determine the temporal variability of the time to maximum aortic density after peripheral venous contrast injection in cardiovascular CT studies and to calculate potential contrast agent savings of selecting scan delay from test bolus kinetics. METHODS: A peripheral contrast bolus was administered intravenously in 138 consecutive patients to determine the time to maximum density in a left ventricular or aortic region of interest by electron beam CT. 20 EKG-triggered scans were acquired within 70 heartbeats. The deviation of the time to maximum density from a standard, fixed scan delay was determined. RESULTS: Mean time to maximum density was 22 s, and the average deviation from that mean was 5 s. At an injection rate of 4 cc/s, this deviation implies that determining the individual scan delay from a test bolus injection may potentially save 20 cc contrast per patient. This amount of contrast agent, in turn, is required for the test bolus. CONCLUSION: Deriving the individual scan delay from test bolus kinetics may improve image contrast in cardiovascular CT studies without additional contrast expense. PMID- 10803046 TI - [Spondylodiscitis with presacral abscesses caused by fistulization in Crohn's disease of the small intestine]. PMID- 10803047 TI - [Double-helical CT pitfall: the native hyperdense basilar artery]. AB - PURPOSE: The unenhanced signs of basilar thrombosis at computed tomography (CT) is the hyperdense visualization of the basilar artery due to intravascular thrombosis. In patients who are clinically asymptomatic, hyperdense visualization of the basilar artery can be observed, if scanned with a double helical CT. Purpose of the present study was to evaluate the diagnostic significance of these changes seen at double-helical CT. MATERIAL AND METHOD: Ten patients patients underwent double-helical CT of a portion of the base of the skull. In each case, 5-mm and 10-mm fused slices were obtained. The patency of the visualized vessels was then documented using contrast-enhanced images. RESULTS: The differences in attenuation between the 5-mm and 10-mm fused slices obtained at native examinations were a median 11 HU. Subsequent contrast enhanced studies documented patency of the examined vessels. CONCLUSION: The present data show that the hyperdense basilar artery as a sign of thrombotic occlusion is not valid when thin, fused slices are obtained at double-helical CT. PMID- 10803048 TI - [Transhepatic duodenal tube in enteral feeding]. PMID- 10803049 TI - [Hyperdense basilar artery in computerized tomography as an indication of basilar thrombosis]. PMID- 10803050 TI - [The biologically equivalent dose (BED): how solid is the calculation of this factor? A consideration of margins of error of biologically equivalent dose]. AB - To predict the effect on tumours in radiotherapy, especially relating to irreversible effects, but also to realize the retrospective assessment the so called L-Q-model is relied on at present. Internal specific organ parameters, such as alpha, beta, gamma, Tp, Tk, and rho, as well as external parameters, so as D, d, n, V, and Vref, were used for determination of the biologically equivalent dose BED. While the external parameters are determinable with small deviations, the internal parameters depend on biological varieties and dispersions: in some cases the lowest value is assumed to be delta = +/- 25%. This margin of error goes on to the biologically equivalent dose by means of the principle of superposition of errors. In some selected cases (lung, kidney, skin, rectum) these margins of error were calculated exemplarily. The input errors especially of the internal parameters cause a mean error delta on the biologically equivalent dose and a dispersion of the single fraction dose d dependent on the organ taking into consideration, of approximately 8-30%. Hence it follows only a very critical and cautious application of those L-Q-algorithms in expert proceedings, and in radiotherapy more experienced based decisions are recommended, instead of acting only upon simple two-dimensional mechanistic ideas. PMID- 10803051 TI - [Diffusion and perfusion: principles and clinical use]. PMID- 10803052 TI - [Radiotherapy of non-malignant diseases: principles and recommendations]. AB - The plenty options and high quality of radiation therapy for non-malignant disorders is not well known outside the field of radiology. It is necessary to transfer this information to cooperating general practitioners, surgeons, orthopedics and other specialists. To warrant quality assurance and quality control and to allow a uniform performance of radiotherapy of non-malignant conditions, general guidelines and recommendations according to the German Working Group of Scientific Medical Societies are useful. This paper summarizes the essential aspects of radiotherapy for non-malignant diseases: indication of, informed consent for, documentation and conduct of radiation therapy for non malignant diseases using orthovoltage equipment and specific recommendations for follow up examinations. Radiotherapy concepts for non-malignant diseases are summarized. PMID- 10803053 TI - [Current imaging in Crohn's disease: value of MRI compared with conventional proceedings]. PMID- 10803054 TI - [A rare complication after foreign body ingestion]. PMID- 10803055 TI - [Prevention of venous thromboembolism]. AB - Deep venous thrombosis and pulmonary embolism, the epidemiologically most important forms of thromboembolic disease still remain one of the most serious problems in medicine. The authors summarize some epidemiological data, the incidence and risk of the disease with regard to the patients condition and the type of insult. The authors mention possible methods of non-specific prevention, types and effectiveness of specific primary and secondary pharmacoprophylaxis and describe physical prophylactic methods. The review offers recommendations for prevention with regard to the assessed risk of the patient and type of insult with special attention to the use of nadroparine. PMID- 10803056 TI - [Femorodistal vascular reconstruction at the Surgical Clinic of the Medical School Hospital in Plzen 1993-1999]. AB - The authors evaluate the success of femorodistal vascular reconstructions during the past seven years, in particular their asset for patients. A total of 41 femorodistal reconstructions were implemented in 39 patients (incl. 25 diabetics- 64.1%). The annual patency of reconstructions is 85.4%, the five-year patency 68.3%. In 11 (28.2%) of patients subsequently a high amputation of the extremity had to be performed. In more than one third of patients with early and late occlusion of the reconstruction it was not necessary to make an amputation and the patients with an originally critical ischaemia of the extremity suffered from brief claudications. The thirty-day mortality was 2.6% (one patient died from embolism of the pulmonary artery). Femorodistal reconstructions have at present a firm position in vascular reconstruction surgery and are usually the last hope for saving the extremity in patients with critical ischaemia of the extremity. They call, however, for exact indication and a delicate surgical procedure. PMID- 10803057 TI - [Quo vadis laparoscopic appendectomy?]. AB - The authors evaluated a group of 282 patients subjected between January 1993 and December 1998 to appendectomy, either by the open or laparoscopic route. They compared the period of work incapacity in the two groups to confirm or rule out the fact that after laparoscopic appendectomy the return to work is faster. The results are more favourable in the group of patients operated by the mini invasive procedure, but even then the period of work incapacity is twice as long, as compared with other countries. PMID- 10803058 TI - [Laparoscopic cholecystectomy in children]. AB - OBJECTIVE: Retrospective evaluation of laparoscopic cholecystectomy in children. The authors deal with the asset of the method in child surgery and emphasize the differences of treatment of cholelithiasis in children by a laparoscopic approach. METHOD: During 1994 to 1998 the authors made 54 laparoscopic cholecystectomies in children 5 to 18 years old (mean age 14.0 years). The indication for surgery was in 48 patients cholecystolithiasis and in six instances confirmed chronic cholecystitis without concrements. Four times cholecystolithiasis associated with congenital haemorrhagic disease was found. The majority of patients, i.e. 51 (94.4%), was operated during the quiescent stage of the disease. RESULTS: In the group of operated children no serious peroperative and postoperative complications were recorded. Twice concrements from the bile duct were removed by papillosphincterotomy in cholecystocholedocholithiasis after previous laparoscopic cholecystectomy. CONCLUSION: In child surgery laparoscopic cholecystectomy is a less frequent operation than in adults. However, it is also in children an effective and safe method with all advantages of mini-invasive operations. PMID- 10803059 TI - [Personal experience with the use of the Powerstar bipolar scissors made by Ethicon]. AB - The authors present their experience with the use of bipolar scissors in surgical practice. They mention the areas of their most frequent use and the advantages of their application. In the discussion they reflect on the economic aspects of operations with this instrument. PMID- 10803060 TI - [Use of a tack for hemorrhage in the presacral plexus]. AB - In operations where the rectum must be mobilized, frequently haemorrhage from presacral venous plexuses occurs. If the haemorrhage is massive, this is a serious complication, which may threaten the patient's life. In those instances we use a "drawing-pin" to arrest haemorrhage. The possibility to use a "drawing pin" is not generally known and therefore the authors present their clinical experience with this method. PMID- 10803061 TI - [Case report of a silent injury of the superficial femoral artery with a late diagnosis]. AB - The authors present a report on almost complete severing of the superficial femoral artery which took a clinically almost inapparent course and the diagnosis was established late. They draw attention to the necessity to consider this type of injury if a foreign body penetrates deeply into Hunter's canal and when the course of the condition is silent. PMID- 10803062 TI - [Use of skin substitutes in treatment of large hypertrophic scars due to burn injury]. AB - The study concerns a clinical experiment in two patients with a dermal substitute based on atelocollagen and hyaluronic acid, and allogeneic acellular dermis. In both cases two-step grafting was performed. At the first step the dermal substitute was implanted into the wound and it was grafted at the second step with thin dermoepidermal autograft. Large hypertrophic scars after burn injury were treated. In both patients the quality of skin cover was significantly improved. PMID- 10803063 TI - [Advances in carotid endarterectomy--review]. AB - Atherosclerosis of carotid artery, resulting in stenosis is a common cause of cerebral ischaemia. Ischaemic stroke is cause of death in 10% of patients and the leading cause of disability in adults. The risk of stroke increases with the degree of stenosis. The diagnosis of the degree of stenosis is performed by duplex sonography, MR-angiography, CT-angiography and conventional angiography. Carotid endarterectomy is a method of choice in the treatment in the case of high grade carotid stenosis. The operation was introduced in 1953. Either regional or general anaesthesia is used for the operation. The different monitoring techniques are used for assessment of the need for shunting. Microsurgical technique enables perfect endarterectomy and fine arterial repair without need for patch grafting. Other techniques of the treatment for carotid stenosis, including carotid angioplasty with or without stenting are subject to the evaluation. Carotid endarterectomy is now the method of choice in the treatment of high-grade carotid stenosis. PMID- 10803065 TI - [History and perspectives in treatment of arterial aneurysms]. AB - The authors give an account of therapeutic trends of arterial aneurysms. As a basis they use a retrospective picture of the history and analysis of the contemporary state of treatment. PMID- 10803064 TI - [Acute pancreatitis--personal experience]. AB - We should like to evaluate in our paper practical experience and diagnostic possibilities as well as treatment of acute pancreatitis in a surgical department of our type and make possible a comparison of our results with those of other departments of a similar or higher type. We also pay detailed attention to the course of the disease and causes of death in patients who died in conjunction with acute pancreatitis in our department. In 1996 to 1998 in our department 39 patients with the diagnosis of acute pancreatitis were hospitalized. This number comprised 28 (72%) men and 11 (28%) women. The mean age of the men was 51.7 years (26 to 88 years), the mean age of women 51 years (18 to 82 years). Five patients (12.8%) died from sequelae of the disease (4 men and 1 woman). Under conditions when examination by a computed tomograph is considered essential, we present experience from a department which does not possess its own apparatus and must make use of capacities of other hospitals. PMID- 10803066 TI - [Traumatic rupture of the descending thoracic aorta. Is emergency surgery necessary?]. AB - The diagnosis of traumatic rupture of the descending thoracic aorta used to be an indication for urgent surgery. Recently, there has been a shift in the perspective regarding the need for an urgent procedure and increasing numbers of surgeons tend to suggest that it is safer to manage first other serious injuries and to perform reconstruction of the injured thoracic aorta in the second stage only. Provided conservative therapy is properly managed, the risk for bleeding from the injured aorta is minimal. In the present case report of a patient with descending thoracic aortae rupture, the authors demonstrate and discuss the strategy of optimal timing of surgery. PMID- 10803067 TI - [Surgical myocardial revascularization in ischemic heart disease in a patient with a transplanted kidney]. AB - The authors presents their first experience with coronary artery surgery in a patient after previous renal transplantation. They describe differences in the preoperative preparation, cardiac operation and postoperative care, with possible risk and complications. PMID- 10803068 TI - [Transhiatal esophagectomy in esophageal carcinoma and subsequent lung carcinoma 6 years later]. AB - The author describes an interesting case primary oesophageal carcinoma. Well differentiated spinocellular carcinoma of the middle third of the oesophagus (T1 N0 MX stage) was diagnosed in a 42-year-old male. He underwent transhiatal oesophagectomy without thoracotomy with a survival period of over 6 year with no relapse. During the last 2 months he developed serious psychiatric and neurological symptoms and neurosurgical craniotomy was performed. The operation presented a metastases of alveolar lung carcinoma in the cerebrum. The primary source was not localised but autopsy revealed a small carcinoma of the left lung with metastases in the cerebellum and basal brain structures causing death. There were no signs of relapse of oesophageal carcinoma during the period after oesophagectomy verified by autopsy. Survival the period after operation was 6 years, 2 months and 2 weeks. Death was caused by subsequent lung carcinoma. PMID- 10803069 TI - [Radical treatment of a primitive neuroectodermal tumor of the thoracic wall]. AB - Primitive neuroectodermal tumours are one of the histological types of soft tissue sarcomas. Although the maximum incidence of these tumours is at a relatively young age, they may be encountered also after the fifth decade. As apparent from the submitted case-record, their clinical and histological diagnosis can be relatively difficult. Treatment is extremely pretentious and the only hope for the patient is radical surgery. After an extensive resection of the thoracic wall the authors decided to replace it by an allograft. The excellent functional and cosmetic results and the relatively long period without signs of a relapse are evidence that the selected procedure was justified. PMID- 10803070 TI - [Secondary hyperparathyroidism in a female patient with six parathyroid glands]. AB - The authors present the case-history of a female patient with chronic renal failure and manifest signs of secondary hyperparathyroidism. The patient was repeatedly subjected to surgical operations during which eventually six hyperplastic parathyroid glands were removed. PMID- 10803071 TI - [Sentinel nodes in breast carcinoma]. AB - The authors summarize literary data on contemporary knowledge and development of views regarding lymphadenectomy in breast cancer. They deal also with the history and technique of detection of a sentinel node by Patentblau and radioisotope 99mTc. In a group of 29 patients operated in 1998-1999 with a detected sentinel node they analyze the surgical technique and present their initial results. In the group they extirpated and examined a total of 354 axillary nodes, i.e. on average 12.2 nodes per patient. Eleven times the nodes were positive, incl. the sentinel node. This number comprised six cases where the sentinel node only was affected. In 16 cases all axillary nodes were negative. In two instances the sentinel node was falsely negative. These results are thus quite comparable with those from reknown departments concerned with sentinel nodes. Examination of the sentinel node very probably determines the affection of axillary nodes and the patient's prognosis. Unnecessary dissection of the axilla with subsequent associated complications can thus be avoided. PMID- 10803072 TI - [Compression anastomosis in acute resection of the colon]. AB - The authors draw attention to the possible use of a biofragmentable anastomotic ring (BAR) for anastomosis of the gut, used by the authors for construction of an anastomosis after acute resection of the colon under particularly risky conditions with infection and advanced deterioration of the intestinal wall. In both instances when this alternative method was used the postoperative course was uneventful and the patients were discharged without complications into domiciliary treatment. PMID- 10803073 TI - [A simple stereotaxic surgical apparatus]. AB - A simple apparatus based on the classical stereotactic method of surgery offers the opportunity to master miniinvasive procedures in neurosurgery and neurotraumatology. This device is particularly helpful in evacuation of intracerebral spontaneous and traumatic haematomas and in stereobiopsies, i.e. the most frequent operations in clinical practice, without the need of general anaesthesia, craniotomy and interference with cerebral tissue. The reliability of the stereotactic apparatus was positively evaluated by three independent surgeons. The apparatus was registered for use in the health services. PMID- 10803074 TI - Ethics-r-us. PMID- 10803075 TI - Ethical decision making: the person in the process. AB - Ethical decisions made by social workers are shaped by the decision maker and the process used to resolve ethical dilemmas. Although systematic guidelines for resolving ethical dilemmas offer social workers a logical approach to the decision-making sequence, it is inevitable that discretionary judgments will condition the ultimate choice of action. Social workers are influenced by professional roles, practice experiences, individualized perspectives, personal preferences, motivations, and attitudes. Through reflective self-awareness social workers can recognize their value preferences and be alert to the ways in which these values unknowingly influence the resolution of ethical dilemmas. Understanding which values or ethical principles were given priority from among competing alternatives can inform social workers about their value patterning. This article challenges social workers to view current ethical decisions as linked to other ethical decisions they have made in the past or will make in the future. An approach to developing keener insight into value patterning is presented. PMID- 10803076 TI - Gandhian principles in social work practice: ethics revisited. AB - Social work as an expression of culture is a highly value-laden activity. The emergence of many new ethical issues resulting from technological and scientific advancements suggests a need for greater attention to values and ethics. In this article the authors argue that the thought of Mahatma Gandhi, as revealed in his social activism, is relevant to social work ethics and a resource for its ethical enrichment. Principles such as seeking truth through service to others, individual self-development, nonviolent social action, and material simplicity could enhance the current NASW Code of Ethics. PMID- 10803077 TI - Sexual contact with clients: assessment of social workers' attitudes and educational preparation. AB - The purpose of this study was to assess social work students' attitudes about sexual contact with clients and their perceptions about their training and education in this area. The sample included 349 social work students in their final semester of an MSW program. There were relatively high levels of approval for sexual contact between social workers and clients in certain circumstances. Approval was not limited to circumstances in which professional relationships were terminated, were brief, or had involved only concrete services. Students with less social work experience and who thought class content on sexual ethics was inadequate were more likely to approve of sexual contact between social worker and client. Students did not feel that they had received adequate education or training on sexual ethics, and many felt unprepared to handle sexual feelings from or toward a client. Implications for education, training, and practice, and suggestions for future research are discussed. PMID- 10803078 TI - Beyond Jones v. Clinton: sexual harassment law and social work. AB - In the wake of the Jones v. Clinton case, there has been a renewed interest in sexual harassment issues, particularly the chaotic case law in this important field of practice. The author urges the social work profession to resurrect its commitment to achieving social justice by participating in the current efforts to unravel the complexities of sexual harassment law. Toward this end, the article presents an analysis of the concept of sexual harassment by providing an overview of the legal issues and case law to highlight the statutory questions that have left the judicial system confounded by the difficulties of practical application of the law. Furthermore, the article deals with the relevance of this topic for the professional mission and suggests the utility of some conceptual frameworks and key concepts that might help social workers to address the clinical, administrative, and advocacy concerns in this substantive area. PMID- 10803079 TI - Ensuring ethical practice: An Examination of NASW Code violations, 1986-97. AB - The NASW Code of Ethics is intended to serve as a guide for practice and as a statement of professional standards that the public may use to hold social workers accountable for their actions. At times, however, the Code's prescriptions may seem overly general, difficult to apply, or unrealistic in light of the challenges workers face daily. How then is the Code interpreted? What actions by social workers may result in findings of ethics violations? This article reviews earlier research on violations of the NASW Code and reports on a recent study reviewing allegations made against NASW members from 1986 to 1997. The article describes the frequency and types of behaviors that resulted in findings of ethical misconduct and offers suggestions for enhancing practice and reducing exposure to ethics complaints. PMID- 10803080 TI - Life after death with dignity: the Oregon experience. AB - This article offers a framework for examination and overview of Oregon's Death with Dignity Act after its first year of implementation. This law became public policy on October 27, 1997, when all legal barriers were lifted. The law allows a terminally ill person to request a prescription to end life if she or he qualifies under the law's requirements. According to records, 23 Oregonians have received such a prescription, and 15 have used it. The effect of physician assisted suicide (PAS) on clients and families, professional health care providers and agencies, and society and culture is explored through two questions: (1) What is known about PAS? and (2) What needs discovery and exploration? The reality of this end-of-life option has forever changed care of terminally ill individuals, and social workers may benefit from the Oregon experience. PMID- 10803081 TI - Protecting the confidentiality of the therapeutic relationship: Jaffee v. Redmond. PMID- 10803082 TI - Advocating for the elderly hard-of-hearing population: the deaf people we ignore. PMID- 10803084 TI - Mirror, mirror on the wall. PMID- 10803083 TI - Economic well-being of children and elderly people. PMID- 10803085 TI - Mirror, mirror on the wall. PMID- 10803086 TI - Outdated practitioner views. PMID- 10803087 TI - Performance of acute flaccid paralysis (AFP) surveillance and incidence of poliomyelitis, 1997-1998 (as of 11 January 1999). PMID- 10803089 TI - Using the health for all framework to explore the development of health policy in the European Region of WHO. PMID- 10803088 TI - Final stages of polio eradication. WHO Western Pacific Region, 1997-1998. PMID- 10803090 TI - The process of developing health for all policy in Finland, 1981-1995. PMID- 10803091 TI - Health policy in North Rhine-Westphalia, Germany. PMID- 10803092 TI - Four variations on one topic: changes in health policy in Hungary (1980-1994). PMID- 10803093 TI - Health policy development in Lithuania. AB - The process of formulating a national health policy in Lithuania, a country in transition, has undergone several stages. Despite the existence of a national critical mass of professionals who understand the major principles of health policy development and who have a solid database to scientifically back the suggested decisions, and despite general acceptance by the public of the broader dimensions of health, the actual process of developing health policy is still facing considerable difficulty. Many factors obstruct this process, such as the inflexibility and resistance of the medical profession, traditions from the previous health service model and an extremely difficult economic situation. Nevertheless, one of the most important factors is the frequent change of government and, subsequently, of ministers of health, which makes continuity difficult. Fortunately, however, in contrast to the political level, the health professionals involved in health policy development and implementation assure at least one level of this necessary continuity. As a result of continuous efforts by the Health Reform Management Group and research and education institutions, and with the constant support of WHO, a policy and strategy for health for all continues to be developed, and the groundwork is being laid for improved dialogue and participation. PMID- 10803094 TI - Health policy development in Malta. PMID- 10803095 TI - Health policy in The Netherlands: description and analysis of ten years of national health policy development emphasizing the health for all strategy. PMID- 10803096 TI - Development of a policy for health for all in Catalonia. PMID- 10803097 TI - Health policy development in Sweden: action at three levels. PMID- 10803098 TI - Comparative analysis of development of health policy in Ostergotland County, Sweden. PMID- 10803099 TI - Healthy city Gothenburg: the work of the Public Health Advisory Committee in evaluating the European regional targets as they relate to Gothenburg. PMID- 10803100 TI - The development of national health policy in Turkey. AB - A written policy is an essential starting point for any administration. Objectives and targets should then be established and strategies to achieve these objectives should be developed. This general approach is also recognized in Turkey, but the policy formulation process is not linked to implementation. The administration does not feel obliged to work according to a plan. Another reason for not adhering to the policy could be that some of the policy principles and targets are too ideal and unrealistic. The targets and objectives are consciously set at an unachievable level to motivate the health workers, but this discourages the health administrators. Services are not evaluated routinely, and the data collected are not processed into usable information and are therefore not used in decision-making. Decisions are usually based on the previous experience of the individuals involved in the policy formulation process or on political will. The high turnover of administrators at the national and provincial levels of the Ministry of Health is another major barrier to adopting and implementing the health policy. If the core of administrators is not stable, a stable policy cannot be established. One of the obstacles to progress was the poor communication between the newly developed Project Coordination Unit and the well established general directorates of the Ministry of Health. Each directorate is autonomous: all deal with different issues. The Unit functions overlapped in many ways with those of the general directorates. This created an awkward situation for them, and sometimes they felt left out from decisions that affected them profoundly. The Unit reports to the undersecretary of state and the minister; therefore, the link with the general directorates was intended to be established through these offices, but sometimes this linkage failed, which resulted in duplication of decisions and activities. Instead of working together towards a common goal, which would strengthen their efforts, they usually work separately. The future is not easy to predict. Change is still needed, but the political environment and level of stability cannot be foreseen. The new Government is pursuing reform activities. The Grand National Assembly has not yet discussed the proposed legislation. New discussions will start at the Assembly, and the policy could be reformulated as a result. This may take some time, which will delay the process. One can hope that the achievements attained so far will be considered and built upon. PMID- 10803101 TI - England: a healthier nation. AB - HINTS AND TIPS: Several difficult challenges have had to be tackled in developing a health policy for England. Although not all the answers have yet been found and the learning process continues, some lessons can be drawn from experience to date. CONSULTATION: Public consultation and the involvement of a wide range of individuals and groups at all levels and stages is crucial to implementing the policy. Without it, The health of the nation would have remained a paper exercise and the local ownership of the policy that has been achieved in some places could not have come about. This principle has been adopted for Our healthier nation, which will benefit from extensive consultation. COMMUNICATION: Communication of the concepts underlying the policy and of ideas about its strategic implementation is also crucial. A wide variety of mechanisms have been used in England, and this has helped to maintain momentum and to keep health policy high on the agenda. Anecdotal evidence suggests that the Health of the Nation calendar and the Target publication have been especially popular. Target in particular has been and continues to be an effective medium for disseminating ideas and examples of successful implementation strategies. In addition, publication of The health of the nation material on the Internet widened its potential audience considerably. TIMING: The publication of The health of the nation was especially timely, not only in terms of gaining support and commitment from the leadership of the Department of Health and other government departments, but also across the political spectrum. In addition, the then-recent NHS reforms gave new opportunities for health policy to be incorporated into health service practice. Our healthier nation is also being launched in tandem with a white paper on health services, and the links between them are being stated explicitly. COMMITMENT: As indicated above, commitment from the top is essential to the success of the strategy; this applies not only to the Department of Health but to all government departments, local authorities, voluntary organizations and others with a role in developing health policy. Such commitment, especially at the local level, has been achieved in part through the fervent efforts of enthusiastic individuals and in part through extensive consultation. CONTENT AND FOCUS OF THE POLICY: The scope of The health of the nation is very wide, but focusing on a limited number of priorities or key areas with challenging but achievable targets has been vital in ensuring progress. In addition, to avoid concentrating exclusively on a small number of challenges, overlying themes such as healthy settings and health alliances allow flexibility and encourage innovation in implementation programmes. INCORPORATION OF EXISTING PROGRAMMES: When the Health of the Nation initiative began, other activities were already taking place locally and within other government departments that could be described as health policy development: for example, at the national level, the Department of Transport's accident targets and, at a local level, the Healthy Cities programmes. One issue that had to be addressed was how to harness these programmes and to bring them under the Health of the Nation banner when there was already strong ownership in other sectors. This was done through communication, consultation and the promotion of intersectoral working. In many cases the pre existing programmes continued as before but with stronger intersectoral links and with a wider health perspective. CONTINUITY: Maintaining continuity in the overall direction of the health policy despite changes in personalities, politically and across the board, has been a challenge and a problem. This has been particularly true at the Department of Health where, as staff tend to move between directorates, corporate knowledge of the process of developing health policy needs to be maintained. (ABSTRACT TRUNCATED) PMID- 10803102 TI - Health gain investment for the 21st century: developments in health for all in Wales. PMID- 10803103 TI - An overview of experience in the European Region. PMID- 10803104 TI - Learning from the past, looking to the future. PMID- 10803106 TI - The effect of flat horseshoes, raised heels and lowered heels on the biomechanics of the equine hoof assessed by finite element analysis (FEA). AB - The biomechanical effects of lowering and raising the heels were studied using a finite element (FE) computer model of the equine hoof capsule consisting of 18,635 finite elements. A static load of 3000 N was distributed to nodes of the inner hoof wall (80%) according to the suspension of the coffin bone, 20% loaded sole and frog. When loaded the FE hoof capsules showed the following deformations: the proximal dorsal wall moves back, the quarters flare to the side and sole and frog perform a downward movement. Stresses are high in the material surrounding the quarter nails, in the heels and in the proximal dorsal wall. Three types of horseshoes were simulated, a regular shoe with flat branches, a shoe with 5 degrees raised heels and a shoe with 5 degrees lowered heels. Raising the heels resulted in significantly (P < 0.05) low stress and displacement values. The lowered heels model calculated highest stress and displacement values and the results of the FE model with the regular horseshoe were found in between. PMID- 10803105 TI - Analgesic, behavioural and cardiopulmonary effects of epidurally injected medetomidine (Domitor) in goats. AB - This study was carried out in order to evaluate the analgesic, sedative, immobilizing and cardiopulmonary effects of medetomidine in goats after lumbosacral epidural injection of three (10, 20 and 30 micrograms/kg body weight) doses. The volume of the injection for all three medetomidine doses was 5 ml in sterile water. Seventeen clinically healthy, Small East African goats of either sex and weighing between 12 and 22 kg (mean +/- SD; 14.8 +/- 2.5 kg body weight) were used. The animals were randomly assigned to two groups. Seven goats were used for evaluating analgesic, behavioural and cardiopulmonary effects while 10 were used for experimental surgery. The cardiopulmonary values and rectal temperature were determined and recorded at time 0 (preinjection) and at 5, 10, 15, 20 and 30 min, and thereafter at 15-min intervals up to 180 min after injection. Analgesia of the flank and perineum was determined at time 0 (preinjection) and at 5, 10, 15, 30, 60, 120 and 180 min using a scoring system. The spread of analgesia to the thorax, neck, forelimbs and head was also determined and recorded. The onset and duration of lateral recumbency was noted and recorded. Medetomidine at the given doses induced variable cardiopulmonary depression, which was not detrimental to the animals. All three doses (10, 20 and 30 micrograms/kg) of medetomidine induced adequate analgesia of the flank and perineum. Analgesia extended to the thorax, forelimbs, neck and head. The duration of lateral recumbency was 136 and 166 min for the 20 and 30 micrograms/kg medetomidine doses, respectively. The duration of lateral recumbency was not determined for the animal given 10 micrograms/kg medetomidine. Signs of sedation (lowering of the head, drooping of the lower lip, partial to complete closure of the eyes and salivation) were noted after administration of all three doses. It can be concluded from this study that all three doses induced adequate analgesia of the flank and perineum. Surgical analgesia of the flank of goats was achieved after lumbosacral epidural administration of 20 micrograms medetomidine/kg, diluted in 5 ml of sterile water. Surgery was not performed with the other doses (10 and 30 micrograms/kg) of medetomidine. PMID- 10803107 TI - Intraperitoneal insemination and retrograde sperm transport in dairy cows. AB - To examine the efficiency of retrograde sperm transport following intraperitoneal insemination, live and dead spermatozoa were used at different concentrations, and sperm recovery from cervical mucus (0.5 ml) 2, 6, 12 and 24 h following insemination was evaluated. Forty lactating Friesian cows, in their second to fourth lactation period, were used in this experiment. Thirty-six cows received intraperitoneally either live or dead spermatozoa. Each group of six cows received one of three total sperm numbers of 30, 45 and 90 million. Four cows were inseminated with 90 million spermatozoa into the uterus and served as a control group. All cows were inseminated towards the end of oestrus. After intrauterine insemination sperm recovery declined, but motile and/or immotile spermatozoa were recovered from all cows at any time. In cows inseminated intraperitoneally, sperm was recovered from the cervix at 6-24 h when 90 million were inseminated. A greater number of spermatozoa was recovered after dead rather than after live sperm inseminations. Only immotile, intact or broken spermatozoa and tail-less heads were recovered after intraperitioneal insemination using either live or dead spermatozoa. No sperm was recovered for 30 and 45 million inseminations. Our results show that, following intraperitoneal insemination, there is passive sperm transport from the peritoneal cavity to the genital tract close to the time of ovulation, and suggest a higher sperm retention in the genital tract when live as opposed to dead spermatozoa are used. PMID- 10803108 TI - Deep-freezing of boar semen in plastic film 'cochettes'. AB - The motility and membrane integrity of spermatozoa from nine boars frozen with a programmable freezing machine in plastic bags, 'cochettes', and in 'maxi-straws', in total doses of 5 x 10(9) spermatozoa/5 ml with glycerol (3%) used as cryoprotectant, were assessed after thawing. A computer-based cell motion analyser was used to evaluate sperm motility, while the integrity of the plasmalemma was assessed with fluorescent supravital dyes (C-FDA/PI). The fertilizing capacity of the semen frozen in the two containers was investigated by inseminating (AI) gilts. Pregnancy was monitored by Doppler-ultrasound, and the numbers of corpora lutea and viable embryos counted at slaughter, between days 30 and 38 after AI. The cochettes sustained the overall procedure of freezing/thawing (FT), with 30 min post-thaw (PT) sperm motility being significantly higher than for straws, 46.9 vs. 39.5%. The only significant difference in motility patterns detected when comparing the packages was a higher sperm velocity (VCL) in cochettes at 30 min PT. However, percentages of FT spermatozoa with intact membranes, detected with the supravital probes, were higher in maxi-straws than in cochettes, 46.8 vs. 43.0% (P < 0.05). There were no significant differences found in fertilizing capacity between spermatozoa frozen in maxi-straws and those frozen in cochettes. The results indicate that although the deep-freezing of AI-doses of boar semen in large plastic bags is feasible, problems such as their inconvenient size for storage and inconsistent thawing must be solved before this type of container can be used for the commercial cryopreservation of boar semen. PMID- 10803109 TI - Evaluation of the role of keratan sulphate as a molecular marker to monitor cartilage metabolism in horses. AB - The role of keratan sulphate (KS) as a metabolic marker of cartilage was evaluated using an in vitro model of equine articular cartilage. Articular cartilage was harvested from clinically healthy 6-month-old foals (n = 3). Chondrocytes were centrifuged and cultured as pellets. Chondrocyte pellets were stimulated by insulin-like growth factor-I alpha (IGF-I alpha) or interleukin-1 alpha (IL-1 alpha) for 2 weeks. The concentrations of sulphated glycosaminoglycans (GAG) and KS in the culture media were measured by a 1,9 dimethyl-methylene blue (DMMB) colorimetric assay and an inhibition enzyme-linked immunosorbent assay using a 1/20/5D4 antibody, respectively. The concentration of GAG was significantly increased both in the media of pellets stimulated by IGF-I alpha and in those stimulated by IL-1 alpha. KS concentration was significantly increased in those stimulated by IL-1 alpha, while no significant change was found in those stimulated by IGF-I alpha. A high correlation between GAG and KS concentrations was found in the media of pellets stimulated by IL-1 alpha (r = 0.84), but not in those stimulated by IGF-I alpha (r = 0.59). The results suggest that the concentration of KS reacting to 1/20/5D4 mirrors the GAG concentration during the stage of cartilage catabolism, but not during the cartilage anabolic stage. The KS concentration in biological fluids could therefore be a useful marker to understand further the cartilage catabolic process. It may also represent some aspects of the cartilage anabolic process. PMID- 10803111 TI - Absence of S100A4 (mts1) gene mutations in various canine and feline tumours. Detection of a polymorphism in feline S100A4 (mts1). AB - Ninety canine and 101 feline tumours of various types were investigated for gene mutations in the coding regions of the metastasis-associated gene S100A4 (mts1). No gene mutations were present in the analysed genomic area. A widespread histidine/tyrosine polymorphism was detected in codon 17 of S100A4. PMID- 10803110 TI - Plasma amino acid pattern during the first month of life in calves fed the first colostrum at 0-2 h or at 24-25 h after birth. AB - Calves are born with a mostly inadequate essential amino acid (EAA) status. Studies were designed to test the hypothesis that delaying the intake of the first colostrum for 24 h, besides its early effects, also has late effects on plasma free amino acid levels and on the protein status. There were marked and rapid elevations (within 2 h) of plasma levels of various amino acids, and especially of EAA, after the intake of the first colostrum, whereas changes after the intake of mature milk on day 28 of life were mostly absent or concentrations even decreased. The EAA and non-essential amino acid (NEAA) status was rapidly normalized after intake of the first colostrum, but normal plasma levels of some amino acids were also reached during the first 24 h of life even when the first meal was withheld. Delaying colostrum intake had only transient effects on EAA and NEAA (except hydroxyproline), in contrast to its effects on plasma immunoglobulin G and total protein levels. PMID- 10803112 TI - Clinical performance of polyacid-modified resin restorations using "softstart polymerization". AB - This study investigated the influence of "softstart-polymerization" on the clinical performance and marginal integrity of polyacid-modified resin restorations (PMR) in class V cavities. Eighty PMR restorations were placed in 20 patients [40 Dyract (DY); 40 Hytac (HY)] with (npat = 10) and without preparation (npat = 10). Restorations were light cured for 40 s either conventionally (CP) or with a lower light intensity for the first 10 s (SSP). Each patient received four restorations (DY-CP, DY-SSP, HY-CP, HY-SSP), which were examined clinically according to modified USPHS criteria, and by quantitative SEM-analysis after 7 days (baseline), 6 months and 1 year. Statistical analysis was performed using the Mann-Whitney-U test (P < or = 0.05) and error rates method. Clinically, no restoration showed recurrent caries or crevices. After 1 year, margins of 24-47% of the HY- and 36-53% of the DY-restorations were rated "Bravo". Marginal discoloration occurred in 20-37% in HY- and in 18-21% in DY-restorations. The error rates method revealed no significant differences between materials or between polymerization modes with and without preparation. Pairwise testing showed that without preparation, the marginal adaptation to dentin was significantly worse compared to enamel for HY with both polymerization modes, for DY with SSP. With preparation, no significant differences were found. Cavity preparation may have an influence on differences in marginal quality between enamel and dentin. PMID- 10803113 TI - In vivo comparison of a microfilled and a hybrid minifilled composite resin in Class III restorations: 2-year follow-up. AB - An in vivo comparison was made of two different types of restorative resins over a 2-year period: a microfilled resin (-1158262462Silux Plus, 3M-1158262462, USA) and a hybrid minifilled composite resin (-1158262461Herculite XRV, Kerr 1158262461, USA); 56 restorations were placed in 28 patients by one experienced dentist and examined by two independent evaluators using the United States Public Health Service (UPSHS) rating system for marginal adaptation, marginal discoloration, surface roughness, anatomic form and modified criteria for color match (direct and indirect evaluation). Modified criteria divided the classic A score into A1 for "not detectable" filling and A2 for "slightly discernible filling". Restorations were evaluated at baseline, 1 week, 6 months, 1 year and 2 years later. After 2 years, all materials were considered satisfactory for marginal adaptation, anatomic form and surface roughness (no "Charlie", or "Delta" ratings). No recurrent caries was observed. Clinical evaluation showed a significantly higher rate of marginal discoloration for the microfilled composite resin than the other resin. Hybrid composite resin materials may be expected to perform well as an anterior restorative material. Photographic ratings confirmed the clinical evaluation. The modified evaluation for color match demonstrated differences, which are not discernible with the USPHS system and showed, more rapidly, differences that appear later with the USPHS system. PMID- 10803114 TI - An evaluation of root canal preparation with the automated Excalibur endodontic handpiece. AB - The aim of this study was to evaluate several parameters for automated root canal preparation with the Excalibur handpiece compared with hand instrumentation. The parameters investigated were straightening of curved canals, cleaning ability, working safety (loss of working length, instrument separation, perforation), cross sectional diameter before and after enlargement, and working time. Sixty curved root canals were prepared using the Excalibur handpiece and 30 curved canals with conventional hand instruments to ISO-size 35. The Excalibur left significantly less debris and smear layer on the root canal walls, but hand preparation resulted in fewer unprepared regions. Root canal curvature was well maintained by either technique; the degree of straightening depended on original root canal curvature rather than on the preparation technique. The Excalibur removed significantly more dentin. In the coronal part of the root canal, hand instruments produced significantly more round and oval cross sections; in the middle and apical part no differences could be detected. Working time was shorter for hand preparation, but the difference was not significant. Manual instrumentation proved to be safe; no instrument fracture, perforation or loss of working length could be observed, whereas automated preparation resulted in one perforation and two cases of loss of working length. PMID- 10803115 TI - Heart transplants--assessment of dental procedures. AB - The object of this study was to evaluate the effects of dental foci on survival rates and rejection episodes in heart transplant recipients. Therefore, in a retrospective longitudinal study we studied 74 heart transplant recipients at the Department of Maxillofacial Surgery and Department of Thoracic and Cardiovascular Surgery, University of Munster. Study patients were divided into groups: those in which dental foci had been verified (n = 31) and those without dental foci (n = 43). Statistical analysis was performed using the chi-square test, Kaplan-Meier life table analysis, and the log-rank test. Before heart transplantation, patients were screened clinically and radiographically to determine the extent of dental foci. Postoperatively, patients were evaluated dentally and medically to identify the impact of dental foci on the incidence of systemic and oral infections, frequency and severity of rejection episodes, mortality, and complications arising during dental treatment. By comparing the mortality, infection and rejection rates in the various groups no statistically significant differences (P > 0.05) were found between patients. Despite immunosuppression, extended inflammatory processes such as abscess formation or viral stomatitis were not found in the oral cavity. We therefore suggest that patients suffering from the symptoms of severe heart failure need not be subjected to rigorous preoperative dental treatment. PMID- 10803116 TI - Helicobacter pylori and ammonia concentrations of whole, parotid and submandibular/sublingual saliva. AB - The aim of this study was to determine the ammonia concentration in whole, parotid and submandibular/sublingual saliva of healthy volunteers using the indophenol direct method. It also investigated the hypothesis that higher saliva ammonia concentrations are associated with the presence of Helicobacter pylori (H. pylori) in the oral cavity. In healthy volunteers, the mean ammonia concentration of whole saliva (2574 mumol/l) was significantly higher (P < 0.0001) than the mean ammonia concentration of both parotid (238 mumol/l) and submandibular/sublingual (355 mumol/l) saliva. In whole saliva, no difference in ammonia concentration was found between healthy controls and dyspeptic patients (mean ammonia values 2574 and 2489 mumol/l respectively, P = 0.7). In addition, no significant differences were observed in the salivary ammonia concentration between dyspeptic patients with and without H. pylori carriage. It is concluded that the ammonia concentration in parotid and submandibular/sublingual saliva does not differ, but is significantly lower than the ammonia concentration of whole saliva. This difference is not due to carriage of H. pylori with its strong urease activity. Therefore, the determination of ammonia in whole saliva is an inappropriate screening test for patients being at risk for (chronic) gastritis and peptic ulcer disease. PMID- 10803117 TI - Ultrastructural investigation of pellicle morphogenesis at two different intraoral sites during a 24-h period. AB - The purpose of the present in vivo study was to examine salivary pellicle formation on enamel surfaces at two different intraoral sites for periods of 1 min up to 24 h by means of transmission electron microscopy. Bovine enamel specimens were attached to the buccal and lingual surfaces of the upper first molars in three subjects using removable intraoral splints. Specimens were carried over period of 1, 10, 30 and 60 min, 2, 6 and 24 h and were processed for transmission electron microscopy. After 1 min, an electron dense pellicle layer, 10-20 nm thick, was observed on the enamel surfaces. The subsequent adsorption of salivary biopolymers was governed by local influences of the oral cavity. Specimens located on the lingual aspect were covered within 2 h by a 20- to 80-nm thick, homogeneous, predominantly granular-structured pellicle. The thickness of the surface coatings that were adsorbed on lingually carried specimens increased to 100-200 nm after 24 h. In contrast, on the buccally mounted specimen surfaces, a variably structured pellicle with granular and globular components could be detected after intraoral exposure for 2 h. The thickness of the 2-h buccal pellicles ranged between 200 and 700 nm. After 24 h, the buccally positioned specimens were covered by a dense globular pellicle layer varying in thickness from 1000 to 1300 nm. It is suggested that in vivo pellicle formation is initiated by adsorption of an electron-dense layer of salivary proteins. Further adsorption of salivary biopolymers leads to the formation of an outer loosely arranged pellicle layer. Under oral conditions, the locally available salivary biopolymers and the influence of locally effective shearing forces are of significance for the ultrastructural pattern and extent of pellicle formation. PMID- 10803118 TI - A comparison of different indices used in the clinical assessment of plaque and gingival bleeding. AB - Indices are used in order to describe clinical variables used in the assessment of periodontal health and diseases. Indices may be divided into those that quantify variables, and those that simply annotate the presence or absence of variables. In this study, quantitative and binary (presence/absence) indices were compared in the evaluation of dental plaque and gingival bleeding. As the data were not directly comparable, they were transformed into percentage changes occurring between consecutive assessments. It was found that the differences between the two types of indices used to assess dental plaque and bleeding on gentle probing were marginal. Thus it was concluded that, the use of quantification indices may have little or no clinical benefit, and did not warrant the extra expenditure of time and effort required for their administration. However, in more detailed clinical and epidemiological trials designed to evaluate the distribution of clinical variables on different regions of tooth surfaces, the quantification indices are necessary. Therefore binary/dichotomous indices may be used effectively, especially in the daily clinical work without recourse to the more detailed and complex quantitative indices. PMID- 10803119 TI - Longevity of cast gold inlays and partial crowns--a retrospective study at a dental school clinic. AB - From 1963 to 1993, 890 patients were treated with 3518 cast gold restorations by students and postgraduate dentists. The longevity of these restorations was studied retrospectively using the patient files. Longevity was calculated using the method described by Kaplan and Meier. After the observation period, 111 (3.2%) of the examined restorations were not in place anymore. The most frequent reasons for failure were caries (33.7%), lack of retention (32.7%), endodontic treatment (29.6%), insufficient marginal adaptation (3.1%) and extraction (1%). The cumulative survival rate and a 95% interval of confidence was calculated for all restorations and for each of the locations and surfaces included in the trial. The 10-year survival rate for occlusal inlays was, 76.1% (12.1) for MO inlays 88.3% (4.2), for DO inlays 83.4% (4.6), for MOD inlays 87.5% (2.4), for partial crowns 86.1% (3.3) and 85.7% (1.7) for all restorations. Based on the statistical method used, the cast gold restorations demonstrated satisfactory longevity results. PMID- 10803120 TI - Identification and characterization of estrogen-like components in commercial resin-based dental restorative materials. AB - Recently, resin-based dental restorative materials have been targeted as potential sources of xenoestrogens, specifically bisphenol A (BPA) and bisphenol A dimethacrylate (BAD), which could contribute to overall estrogen load and result in deleterious side effects. The present study used high-pressure liquid chromatography (HPLC) to analyze twenty-eight different commercially available dental resins for the presence of BPA and/or BAD. In addition, sublines of the MCF-7 human breast tumor cell line were cultured in the presence of eluates from eleven of the dental resins and assessed for proliferative responses using the sulforhodamine B assay. Only one resin, Delton II, had detectable levels of BPA or BAD that could be verified by Fourier transform infrared spectrometry. Likewise, eluates from Delton II were the only samples that elicited a significant proliferative response in two of the MCF-7 sublines tested. Therefore, we conclude that dental resins in general do not represent a significant source of BPA or BAD exposure. PMID- 10803121 TI - Bisphenol-A content of resin monomers and related degradation products. AB - Recently, it was reported that Bisphenol-A (BPA) was released from one fissure sealant (Delton) into saliva causing estrogenic activity in vitro. The aim of this study was to chemically analyze the BPA content of different fissure sealant resin monomers and their release of BPA under hydrolytic conditions. BPA content was first measured in commercially available monomers of bisphenol-A glycidyldimethacrylate (Bis-GMA), bisphenol-A dimethacrylate (Bis-DMA) and bisphenol-A diglycidylether (BADGE). Then, Bis-GMA-monomer and Bis-DMA-monomer in methanol were subjected to pH values of 0 to 11 for 30 minutes at 50 degrees C, to porcine liver esterase, and to pooled saliva for up to 24 hours. The BPA content was determined by high-performance liquid chromatography (HPLC). Bis-GMA monomer and BADGE-monomer from one manufacturer did not contain any detectable amounts of BPA (< or = 2 ppm); Bis-DMA and BADGE-monomer from a second manufacturer contained BPA quantities of 4-155 ppm. For Bis-GMA-monomer, no BPA could be detected under any hydrolytic conditions chosen (detection limit: < or = 1%). For Bis-DMA-monomer an increase of BPA was observed at pH 11, resulting in a conversion of approx. 100% Bis-DMA to BPA. When Bis-DMA was subjected to esterase, a conversion of 82.5% resulted after 24 h; saliva led to an 81.4% conversion of Bis-DMA after 24 h. Hence, we conclude that the results reported in the literature may be attributed to the Bis-DMA-content of the fissure sealant tested (Delton). No BPA-release is expected under physiologic conditions from fissure sealants based on Bis-GMA if pure base monomers are used. PMID- 10803122 TI - Time-related bisphenol-A content and estrogenic activity in saliva samples collected in relation to placement of fissure sealants. AB - It was recently reported that estrogenic activity was detected in saliva samples collected during 1 h after placement of one fissure sealant (Delton) and this related to Bisphenol-A (BPA) content. The aim of the present study was to determine the time-related BPA content and estrogenic activity in saliva samples collected before and after placement of two fissure sealants each with a different monomer composition. Eight healthy male volunteers with no history of prior placement of fissure sealants or composite resin fillings had four molars sealed with either Delton LC (four people) or Visio-Seal (four people). Base-line saliva samples were collected preexperimentally, in the morning when fasting. Fissure sealants were placed and saliva samples collected immediately, 1 h and 24 hs after placement of the fissure sealant. BPA was found in saliva samples collected immediately after placement of Delton LC (range 0.3-2.8 ppm). No detectable amounts of BPA were determined 1 h and 24 h after Delton treatment (detection limit < or = 0.1 ppm). In base-line samples and in all samples collected from Visio-Seal treated individuals, no BPA was detected. In a recombinant yeast cell assay, significantly increased estrogenic activity was found in saliva samples collected immediately after placement of Delton LC sealant (P < 0.05; ANOVA) whereas no statistically significant estrogenic activity was observed in the remaining groups. In conclusion, minute amounts of BPA, however considerably lower than previously reported, were detected in saliva samples collected immediately after but not 1 and 24 h(s) after placement of Delton LC fissure sealant. BPA was not detected after placement of Visio-Seal fissure sealant. PMID- 10803123 TI - Terminal end of the human odontoblast process: a study using SEM and confocal microscopy. AB - Towards the middle of the eighteenth century, Tomes described the presence of membranous structures of cellular origin inside the dentinal tubules. Subsequent studies have been controversial regarding the terminal end of the odontoblasts. According to Fusayama, this cellular process reaches even the dentinal-enamel junction; others, such as Brannstrom, believed that this cellular process is present only in the inner third of the dentin. The aim of the present study was to determine the exact area up to which the terminal ends of the odontoblasts extend. With the aid of advanced confocal laser scanning microscopy (CLSM) cylindrical structures were demonstrated inside the tubules even in the absence of odontoblasts. This would confirm that the structures previously described as cellular processes can be identified with the lamina limitans of the peritubular dentin. High resolution field-emission scanning electron microscopy (FE-SEM) provided further evidence that tubular structures are only seen in the inner third of the dentin, towards the pulp. PMID- 10803124 TI - Activities of lysozyme and salivary peroxidase in unstimulated whole saliva in relation to plaque and gingivitis scores in healthy young males. AB - The purpose of the present investigation was to study the suitability of the salivary activity of lysozyme and salivary peroxidase for monitoring the inflammatory state of the gingiva. Salivary peroxidase and lysozyme activities in resting whole saliva were measured in a group of 140 male subjects (aged 18-30 years). A full mouth, clinical assessment of the plaque index (PI) and the sulcus bleeding index (SBI) was made and gingival crevicular fluid (GCF) flow was measured at teeth 16, 12, 24, 36, 32 and 44 with the Periotron 6000. There were no significant differences in the mean values of lysozyme and salivary peroxidase activities between groups with different PI, SBI and GCF flow values. Statistically significant correlations were found among the clinical parameters, with SBI and PI showing the strongest relation (r = 0.47). The correlation between GCF flow and PI was higher (r = 0.43) than the correlation between GCF flow and SBI (r = 0.20). However, there were no statistically significant correlations between the activities of salivary peroxide and lysozyme and the clinical measures of gingival health. PMID- 10803125 TI - CA 15-3 and c-erbB-2 presence in the saliva of women. AB - Two markers used to aid in the diagnosis and treatment of breast cancer were examined in the saliva of a cohort of 135 healthy women. The investigators detected the presence of cancer antigen 15-3 (CA 15-3) and c-erbB-2 in the saliva sampled from the 135 women. The marker concentrations for CA 15-3 and c-erbB-2 were also evaluated and compared in terms of tobacco usage, menopausal status, estrogen usage, systemic diseases, prescription medications, race, and age. The results of the study showed no association between the aforementioned variables and salivary marker concentrations. The results of this study establish a baseline for measuring the biomarkers in the saliva of women with no evidence of malignant disease and add further support to the notion that salivary concentrations of CA 15-3 and c-erbB-2 may be useful in the detection of breast cancer and/or the post operative follow-up of patients being treated for carcinoma of the breast. PMID- 10803126 TI - In vitro cytotoxicity of resin-containing restorative materials after aging in artificial saliva. AB - Studies have reported that dental resin-based materials release substances which have biological liabilities. However, some current methods for detecting these substances may not be adequate to detect biologically relevant concentrations. In the current study, we hypothesized that resin-based materials exhibit cytotoxic effects and alter cellular function in vitro when high-pressure liquid chromatography (HPLC-UV detection) cannot detect any release of substances. We further hypothesized that this release continues even after aging the samples in artificial saliva. Five types of composite or compomer materials (Z-100, Tetric Ceram, Dyract AP, Solitaire, and Clearfil AP-X) and one organically modified ceramic material (Definite) were tested after aging in artificial saliva for 0, 7, or 14 days. Cytotoxicity was assessed using direct contact with fibroblasts and measurement of succinic dehydrogenase activity after 48 h of exposure post aging. Release of substances from the materials was assessed using HPLC with UV detection. Altered cellular function was estimated by measuring proliferation of MCF-7 cells with sulforhodamine staining. HPLC showed that whereas initial release of substances was higher without aging, this release dropped significantly after 7 or 14 days of aging, and was equivalent to the Teflon controls after 14 days for four of the materials (Tetric Ceram, Definite, Solitaire, and Clearfil AP-X). Without aging in saliva, all materials had cytotoxicities > 50% of the Teflon negative controls. After 14 days of aging, all materials except the Definite continued to show severe cytotoxicity. Only the Definite could be tested for its ability to alter cellular function because of the continuing toxicity of the other materials. This modified ceramic material caused a significant proliferative effect on the MCF-7 cells indicating that sufficient substances were released to alter cellular function. We concluded that all of these commercially available resin-based dental materials continue to release sufficient components to cause lethal effects or alter cellular function in vitro even after 2 weeks of aging in artificial saliva. PMID- 10803127 TI - Effect of toothbrushing on fluoride release of polyacid-modified composite resins. AB - The aim of the study was to evaluate the fluoride release of polyacid-modified composite resins (Dyr-act, Compoglass) submitted to brushing abrasion. Twenty samples were taken from each material and stored in a buffer solution (pH 4.0) for 12 days. Each day, the samples were transferred to a fresh solution. Ten samples of each material were brushed in an automatic tooth-brushing machine (250 strokes, 260-g load) every fourth day. The remaining samples were not subjected to brushing. Fluoride content of the solutions was measured with a fluoride sensitive electrode after the addition of TISAB. Statistical analysis demonstrated significant differences between the two materials with regard to cumulative fluoride release within the 12 days of the experiment. However, no difference was observed between the fluoride release of the brushed samples compared to the unbrushed specimens. This was true for both, the cumulative fluoride release and its release on the day following brushing. It is assumed that regular brushing of the tested materials did not influence their release of fluoride and that brushing of polyacid-modified composite resins does not lead to maintaining their initially high level of fluoride release. PMID- 10803128 TI - Effect of Carisolv solution on sound, demineralized and denatured dentin--an ultrastructural investigation. AB - The purpose of this in vitro study was to analyze the micromorphological changes caused by Carisolv gel on sound, demineralized, and denatured dentin. Fractured dentinal surfaces, dentinal surfaces demineralized superficially by phosphoric acid etching and dentinal surfaces denatured due to lactic acid and collagenase pretreatment were exposed to freshly mixed Carisolv gel or 0.25% sodium hypochlorite. No additional mechanical action was exerted during the 20-min exposure of specimens to the Carisolv solution. Specimens were evaluated by transmission electron microscopy. Electron microscopic evaluation did not indicate any ultrastructural changes of the fractured or demineralized dentinal surfaces due to the 20-min Carisolv treatment. Denatured dentin was partially removed within a 20 min period of chemical action of the Carisolv solution leaving only a 1- to 2-micron thick layer of residual denatured dentin on the specimen's surface. In contrast, 0.25% sodium hypochlorite treatment completely dissolved the demineralized as well as denatured dentin layer within 20 min. It is concluded that Carisolv gel (1) does not affect sound fractured dentin, (2) does not dissolve demineralized dentin, and (3) has a limited potential to chemically dissolve denatured dentin. PMID- 10803129 TI - Absence of an especially toxic clone among isolates of Actinobacillus actinomycetemcomitans recovered from army recruits. AB - Actinobacillus actinomycetemcomitans is a major periodontal pathogen which is associated with early-onset and adult periodontitis. The organism has been shown to be widely distributed among dentate, healthy individuals as well. It has been demonstrated that A. actinomycetemcomitans shows great genetic diversity. An especially virulent clone of the organism (JP2-like) with a specific 530-base pair (bp) deletion in the promoter region of the leukotoxin gene has been isolated from localized juvenile periodontitis patients and related subjects of African and African-American origin. The aim of the present study was to examine the presence of this specific clone employing specific oligonucleotide primers in a polymerase chain reaction among 51 isolates of A. actinomycetemcomitans recovered from 201, 18- to 25-year-old recruits with no or minor periodontal disease. In addition, clinical isolates from 37 periodontitis patients were analyzed as well as reference strains ATCC 29524, NCTC 9710, Y4 and JP2. Primers amplifying a specific 285-bp amplification product in the ltxA region of the leukotoxin gene and a specific 547-bp amplification product of 16 S rRNA were used to genetically confirm identification of the organisms. Primers amplifying sequences in the leukotoxin promoter region were used to identify the deletion. Species specific primers definitively identified all A. actinomycetemcomitans isolates. No isolates from army recruits or the reference strains displayed the deletion in the leukotoxin promoter region. However, in the periodontitis group, a 24-year-old individual from Ghana with localized juvenile periodontitis was identified with an intraoral infection with highly toxic A. actinomycetemcomitans (JP2-like). Present results confirm previous observations of absence of a highly toxic clone of A. actinomycetemcomitans among periodontally healthy and diseased Caucasians as well as a possible presence in localized juvenile periodontitis in individuals of African origin. PMID- 10803130 TI - Root lesions in a group of 50-60 year-old Germans related to clinical and social factors. AB - From a preventive point of view collection of data concerning carious and non carious cervical tooth defects is definitely important. Consequently, the prevalence and distribution of different root lesions were studied and correlated with behavioral and biological factors in 50- to 60-year-old German individuals (n = 298). Additionally, the data were correlated with characteristics concerning oral health and known risk factors such as gender, educational level, and presence of plaque. An interview included questions on sociodemographic and socioeconomic characteristics, dental and general health status, and various behavioral parameters. During clinical examination data concerning coronal and root lesions, restorations, probing depth, gingival bleeding, and dental plaque were obtained. The participants represented a social middle class population with a high awareness of dental health. Obviously, for the participants, known risk factors for root decay such as gender, educational level and plaque index were of minor importance. Factors correlating with root caries were: (a) number of missing teeth, (b) probing depth, (c) smoking habit, (d) regular dental attendance and (e) the reason for the last dental treatment. Additionally, the prevalence of non cariogenic lesions, primarily resulting from increased but wrongly performed oral self care, seems gradually to relieve carious root destruction. PMID- 10803131 TI - Bond strength of a compomer to dentin under various surface conditions. AB - This study determined the influence of different dentin pretreatments on the shear bond strength of an adhesive system and corresponding compomer material. One hundred and twenty freshly extracted human molars were ground with wet SiC paper to expose flat oral/buccal surfaces of superficial dentin. The teeth were assigned to 12 treatment groups (n = 10) based on dentin surface finish (600-grit SiC vs. air abrasion vs. 40 microns diamond bur), surface conditioning (acid etching vs. no etching), and moisture content of the dentin (moist vs. dry). Cylinders of Compoglass F compomer were bonded to the dentin with Syntac Single Component. After 24-h storage in distilled water, the specimens were debonded in shear mode. Bond strengths in MPa (SD) were calculated and bonding sites were analyzed for the mode of failure. Three-way ANOVA revealed significantly higher bond strength values for acid etched specimens (P = 0.001). Moisture content of the dentin surface (P = 0.614) and mechanical surface finish (P = 0.367) had no significant influence on the results. Debonding in unetched groups ranged from 94 to 100% adhesively. Acid etched groups showed adhesive failures ranging from 56 to 100%. To obtain a more reliable bond it is recommended that dentin is acid etched prior to the bonding of a compomer. PMID- 10803132 TI - Cell lipid alterations resulting from prolonged exposure to dimethylaminoethylmethacrylate. AB - Dimethylaminoethylmethacrylate (DMAEMA), a commonly-used component of visible light polymerized dental resins, has the potential to elute and interact with tissue cells to cause cytotoxicity or sublethal metabolic changes. Short-term exposure of cultured oral epithelial cells to sublethal DMAEMA concentrations has been shown previously to affect cell neutral lipid and phospholipid metabolism, resulting in accumulation of significant quantities of dimethylphosphatidylethanolamine (DMPE). In non-treated cells, DMPE is a transient intermediate in phospholipid metabolism and is not detectable by standard methods. In the current study, the effects of prolonged exposure of cells to DMAEMA, and the mechanisms for formation of DMPE in the presence of DMAEMA were examined. Exposure of a keratinizing hamster buccal cheek pouch cell line (HCP cells) to 0.8 mM DMAEMA for 2, 3, 7, and 14 days resulted in reduced incorporation of [14C]acetate into several classes of phospholipids. DMPE was detectable at all time points in DMAEMA-exposed cultures and comprised between 12.48% and 18.33% of the total radiolabeled phospholipids. The results of short term exchange experiments indicated that headgroup exchange was not the major reaction responsible for formation of DMPE in DMAEMA-treated cells; rather the formation appeared to occur through typical phospholipid metabolic pathways. The cells appeared able to re-establish and maintain homeostasis in the presence of this altered cell lipid composition. PMID- 10803133 TI - One year clinical evaluation of the retention and quality of two fluoride releasing sealants. AB - In this prospective, double-blind, split-mouth-designed study, sealant retention rate and quality of two light-cured, fluoride-releasing sealants (Helioseal F, Fissurit F) were compared. Additionally the influence of the isolation method was evaluated. Fifty-eight subjects participated; 1 year later 54 of these (mean age 13.7 +/- 3.6 years) were reexamined. Sealants were applied to 203 tooth pairs (168 conventional and 25 invasive sealants). After 1 year, 193 tooth pairs were available for study; 82 were applied using rubber dam and 111 using cotton roll isolation. Retention rate and sealant quality were judged clinically and with photographs. The initial sealant quality was comparable for both materials. After 1 year, 53.4% of the Helioseal F sealants (44.6% of the Fissurit F sealants) were fully intact, 43.1% (51.8%) partially intact and 3.6% (3.6%) completely lost (P < 0.001). Placement under rubber dam resulted in significantly higher retention rates for both sealants (P < 0.001) as well as in an improved sealant quality. Cases of complete loss only occurred in teeth isolated with cotton rolls. The findings suggest that placement under rubber dam increases retention rate and sealant quality and may reduce material dependent factors that are considered a cause of sealant failures. PMID- 10803134 TI - Elution parameters and HPLC-detection of single components from resin composite. AB - The toxic effects of mercury derived from amalgam fillings have been a focus of controversy in Germany during the last 25 years. As alternatives to amalgam, composites are used widely to restore anterior and posterior permanent teeth. This study quantifies the amount of TEGDMA and methacrylic acid that can be derived from a polymerized commercial hybrid composite material. Different specimens were made with different curing methods with and without an oxygen inhibition layer. To simulate the removal of composite fillings, some samples were pulverized. The specimens were eluted in distilled water at 37 degrees C for different time intervals. To check the influence of acid conditions, pulverized material was incubated in 6 M HCl. The separation of the eluted chemical resin components was done by HPLC. A defined mixture of pure components was used as an external standard for the identification and quantification of the eluted components. The results focus on the release of triethylengly-coldimethacrylate (TEGDMA) and methacrylic acid. Concerning the different incubation methods, the results showed higher TEGDMA elution with oxygen inhibited surface layer and with the pulverized samples. Within acid conditions, the amount of TEGDMA found was low, but the amount of methacrylic acid increased. Small resin monomers like TEGDMA are primarily eluted from composite in aqueous solutions. Within acid conditions a hydrolytic disintegration of the ester bonding of the monomers could be observed. Methacrylic acid--a substance with allergenic potential--could be detected. PMID- 10803135 TI - In vivo forces on oral implants supporting a mandibular overdenture: the influence of attachment system. AB - This study was designed to gain insight into the influence of the attachment system on the loading conditions of oral implants supporting a mandibular overdenture on two implants. Five patients were selected and were provided with two implants in the canine area of the mandible (Branemark System). All patients received a new mandibular overdenture that could be mounted on an ovoid-shaped bar (Dolder, C&M): (a) with and (b) without bilateral extensions and (c) on ball attachments (Nobel Biocare). Using three strain gauges attached to the outer surface of the 5.5-mm standard abutments, the axial forces and bending moments on both supporting implants could be quantified. Load registrations were made during application of 50 N on seven predetermined positions along the occlusal surface of the prosthesis and during maximal biting in maximal occlusion (clenching). The results revealed no differences in induced axial force for the various anchorage devices, unlike the differences in bending moment. Although there is a tendency for better axial load sharing with bars and better sharing of bending moments with ball attachments, these differences were not significant. PMID- 10803136 TI - Internal adaptation and overhang formation of direct Class II resin composite restorations. AB - The aim of the present in vitro study was to evaluate different restorative concepts for posterior resin composite fillings in terms of internal adaptation and overhang formation. Eighty standard occluso-distal cavities with and without a 1.5-mm bevel were restored in a phantom head using Syntac Classic and Tetric Ceram with and without Tetric Flow as thin lining or Solid Bond and Solitaire with and without FlowLine. The restorations were finished intraorally and afterwards subjected to thermal loading (1150 x +5 degrees C/+55 degrees C) for 24 h. The proximal margins of the original specimens were analyzed for overhangs under a stereo light microscope (100-fold magnification) before and after intraoral control with loupes, including additional polish. Afterwards the teeth were cut longitudinally, replicated, and their internal integrity analyzed under a SEM (200-fold magnification). The combination of flowable and viscous composites resulted in enhanced internal adaptation for both adhesive systems. However, Syntac Classic exhibited superior adaptation characteristics compared with Solid Bond. In terms of overhang formation, the use of flowable materials always led to higher percentages of marginal overhangs in beveled cavities. Higher viscous materials alone resulted in higher percentages of underfilled margins of beveled than box-shaped cavities. It was clear that the use of magnifying glasses during finishing was beneficial for reducing marginal overhangs up to 40%. PMID- 10803137 TI - Mercury and silver in saliva from subjects with symptoms self-related to amalgam fillings. AB - The amount of mercury released into saliva from dental amalgam fillings is currently being debated. Mercury enters saliva as vapor, ions and particles of amalgam. The aim of the present study was to determine mercury and silver concentrations in saliva of persons with amalgam fillings. Moreover, it was the aim to investigate whether amalgam particles were present in samples of stimulated saliva in control subjects. In that case, we also wanted to determine the influence of these particles on the mercury concentrations found. Fifty-three patients with a wide range of complaints self-related to their amalgam fillings were examined by the Dental Biomaterials Adverse Reaction Unit of Norway. Among other tests, stimulated saliva was collected from each patient and analyzed for mercury and silver. Mercury and silver correlated with the amount of amalgam present. There was a strong correlation between mercury and silver concentrations. Amalgam particles were found by energy dispersive X-ray analysis. It appears that a considerable part of the mercury and silver were present as amalgam particles. The present study shows that amalgam particles in saliva have to be controlled for when analyzing mercury in saliva from subjects with amalgam fillings. PMID- 10803138 TI - Dental precautions for families with HHT. Hereditary Hemorrhagic Telangiectasia. PMID- 10803139 TI - Dentistry's daddy. The story of G. V. Black. PMID- 10803140 TI - Pathology quiz. White lesions of the gingiva. PMID- 10803141 TI - Detecting and restoring the "hidden" carious lesion. PMID- 10803142 TI - Restoring the age-worn dentition with 21st century technology. PMID- 10803143 TI - The standardized-taper root canal preparation, Part 2: GT file selection and safe handpiece-driven file use. PMID- 10803144 TI - Multipurpose laser offers unique approach to restorative dentistry. PMID- 10803146 TI - Consideration for integrating a dental operating microscope into the general practice. PMID- 10803145 TI - Tips for restoring the implant case. PMID- 10803147 TI - New antimicrobial delivery systems join the fight against periodontal disease. Interview by Phillip Bonner. PMID- 10803148 TI - Preprosthetic cosmetic periodontal surgery, Part 1. AB - There must be constant communication between the periodontist, restorative dentist, and the patient. Detailed techniques must be combined with artistic ideas and tempered with patience. PMID- 10803149 TI - Occlusal and joint theory revisited. PMID- 10803150 TI - Aesthetic materials for the new millennium. Interview by Phillip Bonner. PMID- 10803151 TI - Lower anteriors: the starting point for restoring teeth. PMID- 10803152 TI - A state-of-the-art, multidisciplinary solution to an aesthetic dilemma. PMID- 10803153 TI - Conservative restorations with powerful capabilities, Part 3: Maximizing success with the full-coverage crown. PMID- 10803154 TI - Shaping root canals, Part 4. GT File technique in large root canals with large apical diameters. PMID- 10803155 TI - Nitrous oxide safety: how safe is it for staff? What can be done to make it safer? PMID- 10803156 TI - Aesthetics and dentistry, Part 3: A new technique for cosmetic periodontal surgery. PMID- 10803157 TI - Conservative full-mouth resin renewal. AB - The treatment of this patient involved the coordination of periodontal, orthodontic, restorative, and aesthetic considerations. It was unique because it involved only resin as the primary restorative material, which allowed for conservative preparations and restorations. In the posterior, the use of direct and indirect resins resulted in the removal of little or no healthy tooth structure. In the anterior, the use of direct resin veneers required minimal removal of enamel, while still achieving proper function and aesthetics. Resin restorations in this particular case allowed for an acceptable clinical result and a highly satisfied patient. PMID- 10803158 TI - Taking the mystery out of endodontics, Part 6. Painless anesthesia for the "hot" tooth. AB - Much of what we were taught in dental school is no longer valid. In this article, I've examined some of the myths surrounding what I have dubbed the "inferior" inferior alveolar nerve block. Using the X-tip anesthesia system, it is now possible to achieve total, instant and profound anesthesia of the "hot" tooth, without the problems formerly associated with intraosseous injections. PMID- 10803159 TI - Problem solving in restorative dentistry. PMID- 10803160 TI - Ceramic fractures: cause, repair, and prevention. PMID- 10803161 TI - A simple, rapid, high-quality technique for aesthetic temporization of an anterior laminate veneer or crown. PMID- 10803162 TI - Air abrasion: fixing the cutting edge. AB - The cutting edges of the anterior teeth can be preserved and/or restored using proper concepts and instrumentation. Air abrasion is uniquely well-suited for preparing the class VI restoration. PMID- 10803163 TI - Building the aesthetic practice. Interview by Phillip Bonner. PMID- 10803164 TI - Shaping root canals, Part 3. Large-root canals with small coronal and apical diameters. PMID- 10803165 TI - A three-step procedure for bonding all-ceramic crowns and inlays. AB - To have repeated success with indirect, all-ceramic, resin-bonded restorations, there has to be a consistency in technique. The following are several precautionary steps that should be taken: Get good isolation, preferably by using a rubber dam. However, if that is not possible, use bilateral dry angles, cotton rolls, and suction. Apply primers properly and with the correct amount of time. Air-thin the material. Mix and clean up the cement properly. Develop excellent written communications with the laboratory. Master the technique so that it is easily repeatable. PMID- 10803166 TI - A multiprocedural approach to modern aesthetic dentistry. PMID- 10803167 TI - Coordinating disciplines: aesthetic dentistry and cosmetic surgery. PMID- 10803168 TI - Treatment of missing laterals using tapered implants. PMID- 10803169 TI - Creating exceptional soft tissue contours for fixed anterior bridges and veneers. PMID- 10803170 TI - A technique for fabricating an aesthetic endodontic dowel and metal-free crown. PMID- 10803171 TI - Cosmetic periodontal surgery, Part 2: Using a variations of gingival graft techniques. AB - Fortunately, in this particular case, the patient's dental awareness made it possible for her to request correction of her oral health and aesthetics. This illustration demonstrates the aesthetic awareness and desires of today's society. Practitioners must therefore be able to recognize and work toward these goals. By creatively using variations of techniques to achieve this, the art of dentistry is recovered. Achieving health is primary, but being able to provide a maintainable, healthy, and pleasing appearance is also significantly desirable and important. PMID- 10803172 TI - Biopsy: a new tool for a new age. PMID- 10803173 TI - Microbiologically monitored periodontal therapy. PMID- 10803174 TI - Two new drugs for epilepsy. PMID- 10803176 TI - Substituting for troglitazone (Rezulin). PMID- 10803175 TI - Exemestane for advanced breast cancer. PMID- 10803177 TI - [Beyond 2000, a time to live]. PMID- 10803179 TI - [Message of the President of the 39th SIRM (Societa Italiana de Radiologia Medica) National Congress-2000]. PMID- 10803178 TI - [La Radiologia Medica enters the year 2000]. PMID- 10803180 TI - Diffuse cystic lung disease in the adult patient. PMID- 10803181 TI - [Aneurysms of the abdominal aorta. Guidelines for patient selection and post procedure management. Italian TEAM (Transfemoral Endovascular Aneurysm Management)]. PMID- 10803182 TI - [Gunshot wounds of the cranium studied with computerized tomography. Personal experience in 23 cases]. AB - INTRODUCTION: Gunshot wounds to the head are usually mortal injuries. Their frequency has been increasing in the last years because of increasing crime rates. Gunshot wounds to the head require close clinical and diagnostic cooperation of the neurosurgeon and radiologist, detailed assessment of skull and brain damage, and finally prompt treatment. Emergency Computed Tomography (CT) makes a useful tool for depicting bullet course and brain damage, and thus helps plan treatment. We investigated the CT signs of subdural hematoma, lacerocontusive focus, subarachnoid hemorrhage, hemoencephalus, skull bone fracture and thecal hollow and report them as an aid to the neurosurgeon and the radiologist, for best treatment planning, and in an attempt to establish useful prognostic criteria. MATERIAL AND METHODS: We retrospectively reviewed 23 cases of gunshot injuries to the head studied with CT at the Emergency Unit of Loreto Mare Hospital in Naples, Italy. Twenty patients were men and 3 women; their mean age was 31 years (range: 18-49). Three women and 2 men had been injured accidentally by wandering bullets, and one case was an attempted suicide; all the other cases resulted from shootings. CT slices were 10 mm thick, with 8 mm gap (5 mm in complex injuries and when posterior cranial fossa was involved); all scans were unenhanced. RESULTS: We found 22 penetrating gunshot wounds: 13 of them with thecal entry hole and intracranial bullet retention and 9 with an entry and an exit hole. One case was a superficial wound. Crash skull fractures were seen in 22 cases and they were fragmented in 12, with overlapping thecal fragments in 4, and with deep fragments in 2 cases. There were scattered bone splinters in 3 cases and the bullet was retained in the mastoid bone in one case. Lacerocontusive foci were assessed in 22 cases, brain swelling in 20, subarachnoid hemorrhage in 19, brain hematoma in 15, blood in the ventricular system in 9, pneumoencephalus in 7, air bubbles along the bullet course in 7, impression on ventricula and linear structures in 7, interhemispheric blood in 5, and finally blood effusion in Sylvian scissure in 4 cases. DISCUSSION AND CONCLUSIONS: Gunshot wounds to the head are complex and severe traumas with high mortality rates because of both early and late effects and complications. CT provides the neurosurgeon with abundant findings for diagnosis and surgical planning, which may result in improved survival rates. In these patients emergency CT plays a fundamental diagnostic role in depicting brain damage and thus remains the method of choice for thorough, rapid and accurate brain and skull studies, as well as to detect possible injury to the chest and abdomen. PMID- 10803183 TI - [Usefulness of opposed-phase gradient-echo technique in the diagnosis of occult lesions of the knee and comparison with traditional T1-weight sequences (in phase)]. AB - PURPOSE: To investigate the usefulness of opposed-phase gradient-echo (GRE) technique in detecting occult posttraumatic bone injuries in the knee. Occult injuries account for pain and, if not properly treated, may progress to severe chondral and bone damage. An early diagnosis provided by MRI can help avoid interventional procedures. MATERIAL AND METHODS: We submitted to MRI of the knee 51 patients (32 men and 19 women) with negative plain radiographic findings and at least one traumatic bone injury at MRI. MR examinations were performed with a 0.5 T unit and included a conventional SE or GRE T1-weighted sequence and an opposed phase GRE sequence on the coronal or sagittal plane (2-3 minutes acquisition). To assess the lesion number and conspicuity, images were retrospectively reviewed by two readers. Injury conspicuity was graded as: 0 (poorly visible), 1 (visible), and 2 (well visible). Marrow-to-injury signal intensity ratio was calculated in 30 patients: a ROI was positioned in the site of highest signal intensity and adjacent bone marrow and the ratio analyzed with Student's "t"-test. RESULTS: In-phase and out-of-phase images showed 71 injuries in 51 patients. Conventional (in-phase) imaging missed 6/71 lesions. Injury conspicuity on out-of-phase images was of grade 2 in 58 cases (81.6%) and of grade 1 in 13 cases (18.3%), versus 23 (32.3%) and 42 (59.1%), respectively, on conventional images. Injury conspicuity was graded as 0 in 6 cases (8.4%) on conventional images. Quantitative analysis of marrow-to-injury signal intensity ratio showed higher values for out-of-phase GRE than conventional images. CONCLUSION: Opposed-phase GRE are quick sequences available on all MR systems which appear superior to conventional T1-weighted images in detecting occult injuries in the knee. Injuries are more conspicuous because their signal intensity is lower due to the simultaneous presence of fat and water protons, which is typical of bone trauma, GRE sequences make a useful and rapid complement to T1-/T2-weighted fat saturation acquisitions in the study of the post-traumatic knee. PMID- 10803184 TI - [Os trigonum tarsi syndrome. Role of magnetic resonance]. AB - INTRODUCTION: The os trigonum tarsi is an accessory bone of the foot localized posterolateral to the lateral tubercle of talus. It is usually an asymptomatic condition. However, particular activities such as ballet, soccer, or football may cause repeated stress and chronic microtraumas to the hindfoot, resulting in the os trigonum syndrome. Pain is typically localized anterior to the Achilles tendon; nevertheless, diagnosis may be very difficult because other conditions may show the same symptoms. Radiography can only demonstrate the os trigonum and its morphostructural changes, while MR imaging can also depict associated soft tissue damage. We report on 9 cases of os trigonum tarsi syndrome studied with MR imaging. MATERIAL AND METHODS: Nine patients with the os trigonum tarsi syndrome were submitted to MRI. All the examinations were performed with the patients in supine recumbency with the injured foot in neutral position and then in forced plantar flexion. Axial and sagittal T1 SE, T2* GE and FIR images were acquired. We evaluated os trigonum location and shape, signal intensity of bone, cartilages and adjacent soft tissues, and possible associated tendon injuries. RESULTS: No changes were found in the os trigonum location and shape. Signal intensity changes were seen in 2/9 cases. Particularly, a small area of very high signal intensity, due to necrosis, was depicted on the talar aspect in 1 case; a subchondral spot of slightly increased signal intensity, with a low-signal outline, was seen on the calcanear aspect in another case. Disruption of the cartilaginous synchondrosis between the accessory navicular bone and the posterior tibial aspect was observed in 7/9 patients. Tenosynovitis of the flexor hallucis longus was associated in 6/9 patients. Pseudoarthrosis with irregular bone margins and high-signal spots within the cartilage was found in 3 cases. Finally, fluid effusion surrounding the os trigonum and adjacent soft tissues was always detected. DISCUSSION AND CONCLUSIONS: The os trigonum syndrome may result from chronic microtraumas. Indeed, forced plantar flexion may cause os trigonum compression between the posterior aspect of the tibial malleolus and the calcaneus, with disruption of the synchondrosis with the lateral tubercle of talus. Joint inflammation may be associated with possible development of pseudoarthrosis. Other possible complications are related to vascular changes which may lead to bone necrosis. Furthermore, the particular anatomical site of the os trigonum may sometimes cause compression to the flexor hallucis longus tendon, resulting in severe tenosynovitis. MR imaging allows complete morphostructural assessment because it depicts the margins and the signal intensity of bone and ligaments on the 3 spatial planes. Particularly, sagittal T2 images best demonstrate the cartilage changes indicating synchondrosis disruption. This condition may cause abnormal mobility of the accessory bone with possible impingement with the posterior aspect of the tibia, or hypomobility due to pseudoarthrosis. Forced plantar flexion acquisitions are particularly useful in this condition because they can demonstrate the mechanism of injury. PMID- 10803185 TI - [Pulmonary sequestration in the adult. Diagnostic contribution of angiography]. AB - PURPOSE: The diagnosis of pulmonary sequestration is based on demonstration of mal-developed lung tissue, feeding on abnormal systemic level. We investigate the role of angiography in the diagnosis of pulmonary sequestration in adult patients. MATERIAL AND METHODS: 1987 to 1998 we examined 9 patients with suspected pulmonary sequestration who were subsequently submitted to surgery. The patients were 3 women and men; 6 of them were symptomatic and 3 asymptomatic. Six patients were examined with CT of chest and upper abdomen, thoracoabdominal aortography and selective arteriography of the abnormal vessel; one patient also submitted to left angiopneumography. One patient underwent bronchography and another one MRI. RESULTS: An unquestionable diagnosis was made in 8 cases, namely 6 of intralobar and 2 of extralobar sequestration, and confirmed surgically. The only questionable case was diagnosed at histology as extralobar pulmonary sequestration atypically fed by thin branches from the left diaphragmatic artery. CONCLUSIONS: Angiography demonstrated the abnormal arterial feeding typical of pulmonary sequestration in all cases but one. The evidence of venous drainage was the key sign to diagnose extra-versus intralobar sequestration. Therefore we conclude that angiography remains an essential tool in the diagnosis of pulmonary sequestration, notwithstanding the great potentials of Helical CT of MR angiography. PMID- 10803186 TI - [Diagnostic sensitivity of computerized tomography in closed trauma of the diaphragm. Retrospective study of 35 consecutive cases]. AB - PURPOSE: To evaluate the effectiveness and role of CT in blunt diaphragmatic injuries by reviewing our 8-year experience. MATERIAL AND METHODS: We reviewed the preoperative CT findings of 35 patients with surgically confirmed diaphragmatic rupture. Surgical repair was performed in the acute setting (within 12 hours of trauma) in 22 cases, and late (8 months-5 years) in 13 cases. Twenty eight patients (80%) were examined with conventional CT and 7 (20%) with Helical CT. Scans were initiated at the thoracic inlet to the pubic symphysis, with 8-mm slice thickness, after i.v. contrast agent administration. Four-mm slices were acquired in the region of interest; sagittal and coronal reformations were obtained for Helical CT images. RESULTS: In the acute group, 20 patients had left hemidiaphragmatic and 2 right hemidiaphragmatic rupture; thoracic herniation of the abdominal organs was seen in all cases. Of the 13 patients examined long after trauma, the left hemidiaphragm was ruptured in 12 cases (with visceral herniation in 4), and the right hemidiaphragm in 1, with no herniation. The diaphragmatic rent was found in the dome (15 cases, 43%), musculotendinous junction (11 cases, 31%), muscular portion (8 cases, 23%), and at the muscular attachments on the ribs (1 case, 3%). CT diagnosed diaphragmatic rupture in all the acute cases (22/35 patients, 63%) and in 4 patients with visceral herniation (11%) examined long after trauma. CT findings were questionable in the 9 cases (25%) not presenting visceral herniation. As for the site of diaphragmatic injury, CT never depicted the diaphragmatic rent in the dome and at the musculotendinous junction (74%), not even with thin slices and the multiplanar Helical technique. CT detected indirects signs of injury at the muscular portion (23%), showing the injury site directly in the case with diaphragmatic avulsion (3%). CONCLUSION: CT is a reliable tool in the diagnosis of suspected diaphragmatic injury in the acute trauma setting. Long after trauma, CT performs poorly because it depicts the diaphragmatic rent only in some peripheral traumas. Helical CT has greater diagnostic potentials, but the injury site and type do affect its capabilities. PMID- 10803187 TI - [Carbon dioxide angiography. A new injection system]. AB - PURPOSE: To evaluate the usefulness of a new carbon dioxide (CO2) intravascular injection system in digital subtraction angiography. MATERIAL AND METHODS: March 1998 to May 1999, thirty-nine patients were submitted to digital subtraction angiography with CO2 injection by a new delivery system, CO2-Angioset, OptiMed, Ettingen, Germany. The patients were 29 men and 10 women, whose age ranged 32 to 76 (mean: 47), eighteen of them with absolute or relative contraindications to iodinated contrast media and 6 with poor diagnostic findings at previous conventional angiography. CO2 was used for comparison with iodinated contrast agents in 4 patients. We studied the following vascular districts: renal arteries in 9 patients, portal vein in 18, lower limb arteries in 7, upper limb veins in 4. In 1 patient CO2 angiography was carried out for the diagnosis and interventional treatment, by transcatheter embolization, of a postbioptic arteriovenous renal fistula. During the procedure, arterial blood pressure, EKG status and oxygen stauration were monitored, and subjective sensations recorded in all patients. RESULTS: CO2 angiography provided adequate visualization of vascular districts and of abnormal findings in 32 cases (82%), while its results were considered insufficient for correct and complete assessment in 7 cases (18%). In detail, renal arteries studies were adequate in 9/9 cases, providing good depiction of the arterial trunk and main branches but poor demonstration of interlobar and arcuate arteries. The portal trunk was well depicted in 12/18 cases, but CO2 angiography results were insufficient in 3 cases because of poor catheter wedging in the hepatic vein and in 2 cases because of CO2 drainage by an accessory hepatic vein. Peripheral circulation was clearly depicted in 5/7 cases, while there was incomplete filling of the abdominal aorta and of iliac and femoral arteries in 2 cases. We had only one transient complication (2.57%) due to mesenteric ischemia during an aortic injection, with mild abdominal pain and diarrhea which subsided spontaneously in few minutes. CONCLUSIONS: In our experience the CO2-Angioset delivery system has proved to be a simple and safe tool, particularly suitable for use in patients at risk for allergic reactions to iodinated contrast agents and in those with renal function impairment. Also, the system can help carry out some interventional procedures such as arteriovenous fistula embolization and transjugular portosystemic shunting. PMID- 10803188 TI - [Gastric stromal tumors. Findings with computerized tomography]. AB - PURPOSE: Gastric stromal tumors are an ill-defined group of lesions arising from muscle wall cells and characterized by extremely variable biological patterns. Thanks to modern immunohistochemical and ultrastructural techniques, four main classes of these lesions have been identified, namely: 1) tumors with differentiation toward smooth muscle cells; 2) tumors with differentiation toward neural elements; 3) tumors with dual differentiation toward both cell types; 4) tumors lacking differentiation toward either cell type. We investigated the yield of CT in diagnosing and characterizing gastric stromal tumors. MATERIAL AND METHODS: We retrospectively reviewed the CT findings of 38 patients (15 men and 23 women; mean age 51 years) with pathologically proven gastric stromal tumors, namely 31 of myoid origin, 4 of neural origin, 2 with both muscle and neural differentiation, 1 lacking differentiation with either cell type. Morphological (size, margins, growth pattern, enlarged lymph nodes, metastases) and structural (density, calcifications, necrosis, cystic changes, enhancement patterns) parameters were evaluated and compared with histopathologic diagnosis. RESULTS: The myoid tumors involved gastric fundus in 9/13 and 5/7 benign lesions and lesions with variable biological patterns, respectively, while the malignant tumors exhibited diffuse involvement in 7/11 of cases. Eleven of 13 benign lesions had regular shape and a diameter +/- 5 cm, while 5/7 intermediate tumors and 7/11 malignant ones were 6-10 cm and over 10 cm (1/7 and 3/11, respectively). Tumor growth was intramural (6/13) or mixed (5/13) in the benign lesions, mainly exophytic in the malignant ones, and finally variable (2/7 intramural, 3/7 exophytic and 2/7 mixed) in the tumors with intermediate patterns. Calcifications were identified only in 3 benign lesions. Structure was homogeneous in 11/13 of the benign lesions with regular contrast enhancement (10/12), heterogeneous due to necrotic areas in nearly all (10/11) or most (5/7) malignant and intermediate forms, respectively. Enlarged lymph nodes (4/11) or synchronous metastases (7/11) were found in the malignant lesions. Our two schwannomas had a pseudocystic structure with regular peripheral contrast enhancement; the neurofibroma was characterized by multiple, markedly hypodense and hypovascular lesions. Both the malignant neurogen form and the two tumors with dual differentiation showed a diffuse, aspecific and infiltrating pattern. The lesion without differentiation had a "benign" shape and structure even though it developed metastases two years after radical surgery. CONCLUSIONS: Despite the lack of specificity of CT patterns of gastric stromal tumors, these parameters are useful in differentiating benign from overtly malignant forms. The definition "gastric stromal tumor with benign, variable, or malignant macroscopic appearance" should be used in the radiologic report. PMID- 10803189 TI - [Personal experience in 71 consecutive patients with acute cholecystitis]. AB - PURPOSE: Acute cholecystitis is one of the most frequent abdominal inflammatory processes. If untreated or misdiagnosed it can result in severe complications such as gallbladder rupture, abscesses, or peritonitis. We retrospectively reviewed a series of 71 consecutive patients with surgical confirmation of acute cholecystitis and now compare the results of the diagnostic techniques we used preoperatively. MATERIAL AND METHODS: Over 16 months, 71 consecutive patients (42 women and 29 men; age range: 34-84 years, mean: 58) with acute abdominal pain were operated on for acute cholecystitis at Cardarelli Hospital, Naples. Abdominal plain film was performed in 65 of 71 cases, abdominal US in 69 and abdominal CT in 6. On abdominal plain films, we retrospectively searched the following signs: densities projected over the gallbladder, linear calcifications in gallbladder walls, gallbladder enlargement, focal gas collections within the gallbladder, and air-fluid levels in the gallbladder lumen. On US images we looked for: gallbladder wall thickening (> 3 mm), intraluminal content in the gallbladder, pericholecystic fluid, US Murphy's sign, and gallbladder distension. On CT images, we investigated: gallbladder distension, wall thickening, intraluminal content, pericholecystic fluid, and inflammatory changes in pericholecystic fat. Associated complications of cholecystitis were also searched on all images. RESULTS: On plain abdominal films we found densities projected over the gallbladder (16.9%) and linear calcifications in the gallbladder wall (4.6%). Abdominal US demonstrated gallbladder wall thickening (56.5%), one or more gallstone(s) (85.5%), pericholecystic fluid (14.5%), gallbladder distension (46.4%), and US Murphy's sign (39.1%). Abdominal CT showed gallbladder wall thickening (83.3%), gallbladder distension (66.6%), pericholecystic fluid (66.6%), gallstones (50%), inflammatory changes in pericholecystic fat (33.3%), and increased bile density (> 20 HU) (33.3%). CONCLUSIONS: US appears to be the most useful imaging technique in patients with suspected acute cholecystitis, for both screening and final diagnosis. CT plays a limited role in the early assessment of these patients, but can be a useful tool in diagnosing acute cholecystitis in patients with questionable physical findings or in investigating related complications. PMID- 10803190 TI - [Echographic study with high-frequency and high-spatial resolution transducer in the evaluation of renal transplant in pediatric age]. AB - PURPOSE: We investigated the role of power Doppler US with a high-frequency and high-resolution transducer (13 MHz) in the visualization of interlobular arterioles in patients with normally functioning renal transplants or with chronic rejection. MATERIAL AND METHODS: We examined 15 patients (mean age 15 years; range 10-18 years) with a General Electric 500 MD unit using 7.5 and 13 MHz linear transducers. In all the patients serum creatinine and diuresis were evaluated; 4 patients underwent US-guided biopsy that resulted in the diagnosis of chronic rejection. RESULTS: Normally functioning renal transplants were found in 11 patients and chronic rejection was seen in 4. In normally functioning renal transplants, interlobular vessels could be depicted as "cortical blush" with the 7.5 MHz transducer; in the same patients power Doppler US with the 13 MHz transducer permitted a correct evaluation of interlobular vessels that were arranged in series like a palisade. In chronic rejection power Doppler US with the 13 MHz transducer better depicted cortical vascularity and showed irregular, narrow arteries. CONCLUSION: Power Doppler US with a 13 MHz transducer is particularly useful in children after renal transplants due to their reduced tissutal thickness. The lateral resolution of 13 MHz transducers (< 0.3 mm) allows to separate interlobular vessels from each other and the high frequency of the probe can depict interlobular vessels in the peripheral cortex. The optimal visualization of cortical vascularity with a 13 MHz transducer allows early detection of chronic rejection. PMID- 10803191 TI - [Computerized tomography in the study of the diabetic kidney]. AB - PURPOSE: We investigated whether any morphometric or densitometric changes are detectable in the kidneys of type 2 diabetic patients. MATERIAL AND METHODS: We retrospectively reviewed the CT findings of 40 diabetic patients and of 20 non diabetics (the control group) submitted to triphasic helical CT for different abdominal conditions. The type 2 diabetic patients were 23 men and 17 women, mean age 62 years, while the nondiabetic controls were 12 men and 8 women, mean age 58 years. All the CT images were analyzed using the Multiplanar Reconstruction (MPR) software. We classified the diabetic patients as either nephropathic or non nephropathic based on laboratory signs of renal disease, and divided them into three groups based on the duration of their diabetes (0-5 years, 5-10 years, > 10 years). We evaluated morphometric features, such as the presence of parenchymal or vascular calcifications, axial and coronal renal diameters, coronal renal area, and corticomedullary ratio, and densitometric and functional features, such as unenhanced renal density, cortical and medullary density in the arterial phase, parenchymal density in the nephrographic phase, and contrast elimination. We also compared the results in our subgroups of patients with those in the controls. RESULTS: Both renal diameters on axial sections and cortical density in the arterial phase were significantly lower in the nephropathic group (p < 0.05) than in nonnephropathic patients and controls. The patients with shorter duration of their diabetes had significantly greater axial diameters (p < 0.05); a longer standing disease correlated significantly (p < 0.05) with smaller axial diameters of kidneys and lower cortical density in the arterial phase. No significant correlation was found for any other parameter. CONCLUSIONS: Renal diameters on axial and coronal sections and cortical density in the arterial phase can be useful indices of early nephropathy in diabetic patients. Further studies are warranted to make these findings suitable for use in clinical practice. PMID- 10803192 TI - [Multiple prostatic biopsies in the "gray zone" of the specific prostatic antigen]. AB - PURPOSE: Aim of our study was to identify cases of undetected prostatic cancer in patients with normal findings at digital examination and transrectal US, and prostate specific antigen (PSA) values ranging 4-10 ng/mL. MATERIAL AND METHODS: Two hundred and ninety patients were submitted to transrectal US and random bilateral prostatic biopsy; 3 samples were collected from each side of the gland using 16-Gauge thru-cut needles. Of the 290 patients who gave fully informed consent, we selected 34 whose age ranged 56 to 76 years (mean: 64). Inclusion criteria were PSA 4-10 ng/mL, PSAD cut-off 0.15, free/total PSA ratio 15-25%, and normal findings at digital examination and transrectal US. PSA velocity was calculated collecting 3 blood samples every 30 days for 2 months. RESULTS: Five of the 34 selected patients (15%) had prostatic cancer, and 2 (6%) Pin (1 Pin 1 and 1 Pin 2). As for the other 27 patients, biopsy demonstrated 4 (12%) cases of prostatitis and 23 (62%) cases of BPH. PSA values increased in all patients with positive histology, versus only 6 (22%) of those with negative histology. PSAD was 0.15 or greater in 3 of 7 prostatic cancer patients. Free/total PSA ratio never exceeded the cut-off value. Gleason score ranged 2 to 4. CONCLUSIONS: Our findings confirm that prostatic biopsy can detect tumors also in areas which appear normal at transrectal US and digital examination, and that PSA rate increases in patients with positive histology. Finally, the actual clinical role of prostatic biopsy relative to all other diagnostic imaging techniques remains to be defined. PMID- 10803193 TI - [Intraosseous lipoma of the calcaneus. Characterization with computerized tomography and magnetic resonance in a case]. PMID- 10803194 TI - [Meyer's femoral dysplasia. Description of a case]. PMID- 10803195 TI - [Skeletal radiologic aspects of 2 rare cases of porphyria]. PMID- 10803196 TI - [Sister Mary Joseph's nodule from a metastatic carcinoma. Description of 2 cases]. PMID- 10803197 TI - [Poland syndrome. Description of a case]. PMID- 10803198 TI - [Pseudoneoplastic mammary nodule: sarcoidosis]. PMID- 10803199 TI - [Thrombosis of the abdominal aorta and splenic artery in antiphospholipid syndrome. Description of a case studied with computerized tomography]. PMID- 10803200 TI - [Thrombosis of the common trunk, decreased portal blood flow, and consequence on hepatic perfusion. Considerations on 2 cases evaluated with spiral computerized tomography]. PMID- 10803201 TI - [New computerized tomography findings in infarct of the entire colon: gas in the inferior mesenteric vein, the parietal branches and the visceral origin of the plexus of the internal iliac veins. Description of a case]. PMID- 10803202 TI - [Peritoneal pseudocysts. Presentation of 4 cases]. PMID- 10803203 TI - [A rare case of granulomatous appendicitis]. PMID- 10803204 TI - [A rare case of splenoma studied with several diagnostic imaging methods]. PMID- 10803205 TI - [Pancreatic echinococcosis. Computed tomography aspects in a case]. PMID- 10803206 TI - [Acetylcholine receptor knockout mice]. AB - To identify the functions of nicotinic or muscarinic acetylcholine receptor (nAChR or mAChR) subtypes, mice lacking beta 2 nAChR, alpha 4 nAChR, alpha 7 nAChR, M1 mAChR, and M2 mAChR have been generated. All these mice grow to normal size, and show no obvious physical or neurological deficit. However, pharmacological, biochemical, electrophysiological, neuroanatomical, and behavioural analyses revealed important functions of these AChR subunits. The beta 2 nAChR is most widely expressed in the central nervous system, and is involved in the functional high-affinity nicotine receptor regulating cognitive performance and the mesolimbic dopamine system. Aged beta 2-/- mutant mice showed neocortical degeneration and impaired spatial learning, and may serve as one possible animal model for dementias. The alpha 4 nAChR is associated mainly with the beta 2 subunit, and may form a component of the nicotinic pain pathways modulating the antinociceptive effect of nicotine. The alpha 7 nAChR mediates fast nicotinic currents in the hippocampus, and is not essential for normal neuronal development nor neurological function. The M1 mAChR mediates M current modulation in sympathetic neurons and the induction of seizure activity in the pilocarpine model of epilepsy. The M2 mAChR functions in the extrapyramidal system, hypothalamus, and spinal and/or spraspinal muscarinic pain pathways, and is possibly involved in locomotor performance, temperature control, and antinociceptive responses, respectively. PMID- 10803207 TI - [Opioid receptor knockout mice]. AB - mu, delta, kappa opioid receptors are target molecules for analgesia, reward and many physiological functions of opiates. Opioid receptor knockout mice generated by gene-targeting technology which can introduce mutation into specified locus provide invaluable animal models to elucidate the in vivo function of opiates and develop new therapeutic drugs. The disruptions of mu receptor expression decreases the nociceptive threshold to thermal stimuli and increases the threshold to visceral chemical stimuli paradoxically. Analgesia, reward, respiratory depression, constipation, immunosuppression and physical dependence induced by morphine are absent in mice lacking the mu receptor. These data show that the mu receptor is a molecular target for most effects of morphine, both therapeutic and side effects. mu Receptor expression is required for most delta receptor-mediated and some kappa receptor analgesic effects. These results support substantial roles for mu receptor in the analgesic properties of delta, kappa receptors. Cocaine and ethanol reward require mu receptor systems' intactness. Mice lacking the mu receptor will be a useful tool to study complex interactions between endogenous opiate and dopamine systems. PMID- 10803208 TI - [Dopamine receptor knockout mice]. AB - Dopaminergic systems are transmitted by dopamine receptors which couple to GTP binding proteins. Five subtypes of dopamine receptors were so far cloned. To study the functions of each dopamine receptors, mice lacking each of D1R, D2R, D3R and D4R dopamine receptors have been generated. Histological analyses of D1R knockout mice indicated that the expression of dynorphin is reduced in the striatum. In contrast with wild-type mice, D1R knockout mice exhibit a dose dependent decrease in locomotion. D2R knockout mice display a hypoactivity. The expression of enkephalin mRNA in the striatum is increased in the D2R knockout mice. D2R knockout mice showed hyperplastic changes of intermediate lobe of the pituitary and the increased expression of POMC in the pituitary in D2R knockout mice. D3R knockout mice are more active than wild-type mice in a novel environment and they exhibit enhanced behavioral sensitivity to cocaine and amphetamine. D4R knockout mice show a hypoactivity, but they display locomotor supersensitivity to ethanol, cocaine, and methamphetamine. Dopamine synthesis and its conversion to DOPAC are elevated in the dorsal striatum from D4R knockout mice. PMID- 10803209 TI - [Serotonin receptor knockout mice]. AB - The neurotransmitter serotonin is known to play a key role in determining emotional state in higher animals, especially in humans. Pharmacological studies have highlighted a subset of serotonin receptor subtypes (1A, 2B and 2C) worthy of knockout analysis. Several groups investigated knockout phenotype and determined the physiological function of these receptors, although the phenotype of the mouse seems somewhat paradoxical. However, the results strongly suggest that serotonin receptors are mainly involved in emotion (i.e., anxiety, aggressive behavior, obesity and epilepsy). PMID- 10803210 TI - Quantification of neuromodulin (GAP-43, B-50) and synapsin I in rat striata. AB - We reported previously that phosphorylated neuromodulin and phosphorylated synapsin I content increased in the striata of amphetamine-sensitized rats; however, the neuronal pathways responsible for the increase were unclear. In the present study, changes in neuromodulin and synapsin I content resulting from the manipulation of lesions were quantified to elucidate the responsible pathways. Nerve terminals originating in the corticostriatal pathway, those from the nigrostriatal pathway and those from interneurons in the striatum, were impaired by unilateral cortical ablation, 6-hydroxydopamine (6-OHDA) treatment and kainic acid injection into the striatum, respectively. Neuromodulin and synapsin I content in the ipsilateral striatum after unilateral ablation of the frontal cortex decreased by 51 and 31%, respectively. The impairment of dopaminergic terminals by 6-OHDA reduced the neuromodulin content by 22%; however, no significant alteration was observed in the synapsin I content as the result of 6 OHDA treatment. The injection of kainic acid did not cause the content of either protein to decrease. These results suggest that corticostriatal nerve terminals possess a large part of the total neuromodulin and almost all the synapsin I in the striatum. Therefore, the increase in phosphorylated neuromodulin induced by repeated treatment with amphetamine may occur in corticostriatal glutamatergic terminas and/or nigrostriatal dopaminergic terminals. On the other hand, the increase in phosphorylated synapsin I may preferentially occur in the corticostriatal glutamatergic terminals. PMID- 10803211 TI - Cerebral acetylcholine levels and long-term spatial cognitive impairment following middle cerebral artery occlusion in rats. AB - The behavioral and neurochemical changes in the chronic phase of permanent occlusion of the right middle cerebral artery (MCA) in rats were investigated. Nineteen MCA-occluded rats failed to solve the 8-arm radial maze task (cognitively impaired rats), while 11 MCA-occluded rats could complete it (cognitively unimpaired rats). When a delay of 60 min was imposed in the task, however, 5 cognitively unimpaired rats failed to complete the task. The rats that underwent behavioral testing were studied for any changes in ACh levels in various brain regions using HPLC with electrochemical detection. The ACh levels in the infarcted areas decreased considerably in all MCA-occluded rats, but no region of the infarcted areas correlated with the spatial cognitive deficit. The ACh levels tended to decrease in the frontal cortex of the cognitively impaired rats and greatly increased in both ipsilateral and contralateral parietal cortex of the cognitively unimpaired rats. A significant correlation was observed between the ACh levels and spatial cognitive deficit in the contralateral frontal cortex, and ipsilateral and contralateral parietal cortex. These results suggest that the cholinergic function of the frontal and parietal cortices might play a role in acquiring spatial cognition in MCA-occluded rats. PMID- 10803212 TI - [Relationship between hippocampal arachidonic acid content and induction of LTP in aged rats]. AB - There are several lines of evidence indicating that membrane AA correlates with the ability of aged rats to sustain LTP. The age-related decrease in membrane AA seems to be triggered by increased lipid peroxidation, which is involved with the decline of LTP. The chronic treatment of DHA could decrease membrane AA without an increase in lipid peroxidation. We have thus investigated the effect of chronic treatment of DHA on hippocampal LTP to assess whether the decrease in membrane AA could directly affect the induction of LTP. The effects of daily supplementation of DHA for 3 months on membrane AA, LTP, and Ca2+ response were evaluated using hippocampal slices from 26-month-old Wistar rats. Chronic treatment of DHA reduced the hippocampal AA significantly, but did not change the amplitude of LTP. Neither 30 mM K+ nor 500 microM NMDA-induced Ca2+ response was affected by chronic treatment of DHA, while the 500 microM carbachol-induced Ca2+ response was reduced. From these results, the reduction of membrane AA might suppress the carbachol-induced Ca2+ response by inhibiting the muscarinic receptor function, IP3 formation and/or Ca2+ release from Ca2+ stores by IP3. However, the reduction of membrane AA is not likely to be a main causal factor of the decline of LTP. PMID- 10803213 TI - [Significance of public health regulation for quality of products of aerobic and anaerobic biological waste processing]. PMID- 10803214 TI - [Effect of plant diseases and plant pests on phytohygienic safety of compost]. AB - During the last years the production of compost made from separately collected organic waste heavily increased (about 7.5 mio tons) and is supposed to continue increasing. For the use of those products in crop cultivation high demands on the quality and especially on the freedom from phytopathogenic organisms has to be met. Regarding investigations published up to now on the ability of plant seeds and phytopathogenic organisms to survive the decomposition process, it can be deduced that composting under optimal conditions usually may ensure substantial freedom from phytopathogenic organisms. There remains the need for further research as to the stability of heat-resistant viruses, of viruses with fungal vectors and of quarantine organisms. Data on the effect of anaerobic digestion to phytopathogens are also lacking. A monitoring program to check composts produced on an industrial scale for freedom from phytopathogenic organisms is proposed. Some provisions of the German 'Bioabfallverordnung' dealing with phytosanitary aspects are critically discussed. PMID- 10803215 TI - [Biological waste regulation--a means of quality assurance for production and use of compost products]. PMID- 10803216 TI - [Quality assurance measures and and design evaluation of the RAL Compost Quality Assurance--self control measures of refuse site managers]. PMID- 10803217 TI - [Communicable disease control evaluation of anaerobic biological waste treatment in comparison with aerobic composting]. PMID- 10803218 TI - [Use of compost products from the viewpoint of agriculture]. PMID- 10803219 TI - An EU research project on waste handling. PMID- 10803220 TI - An overview of waste management in the United States and recent research activities about composting related occupational health risk. PMID- 10803221 TI - [Immunological reactions and health complaints in biological refuse personnel and composting by biological aerosol exposure]. AB - Employees in waste treatment industries are exposed to high levels of airborne microorganisms. Concentrations of 10(7) cfu/m3 air were measured in composting facilities, whereas exposures of bio-waste collectors were two orders of magnitude lower with exposure peaks up to 10(6) cfu/m3 air. These exposures were compared to specific antibody levels against molds and actinomycetes, work related health complaints, and diseases of the workers. 72 bio-waste collectors and 77 compost workers were examined and interviewed for work related symptoms, current occupational exposures, smoking habits, and exposures from other sources (farming, bird breeding, gardening) by physicians specialized in occupational medicine, using a standardized protocol. Levels of specific IgG antibodies to molds and actinomycetes occurring regularly at these workplaces were determined as biomarkers of exposure. The exposure data from the workplaces indicate a higher exposure of compost workers compared to bio-waste collectors. This corresponds with significantly increased health complaints of the upper airways and the skin and higher concentrations of specific antibodies against molds and actinomycetes in the compost workers. Atopic diseases were underrepresented in the compost workers compared to bio-waste collectors and population based data, indicating a healthy worker effect in this group. Occupational diseases from airborne microorganisms eligible for insurance compensation were observed in waste treatment facilities. More detailed studies are needed for exact assessment of health risks of workers in these industries. PMID- 10803222 TI - [Mycotoxins as exposure parameters in bioaerosols of composting sites]. AB - The potential to produce mycotoxins was investigated for freshly isolated strains of airborne fungi. The spectra of metabolites in conidial extracts and culture extracts were compared for some relevant species. Furthermore, their potential to produce mycotoxins on semi-natural media (compost extract agar) supplemented with sucrose, yeast extract, and carboxymethylcellulose in different combinations was investigated. In native bioaerosols in a compost facility (plant 2), tryptoquivaline, a compound with tremorgenic properties, and trypacidin, for which no toxic properties are described, were found. The highly toxic metabolites gliotoxin and verruculogen were not found in the bioaerosols, although they were produced by some strains in pure culture. An inventory of microbial metabolites in addition of fungal propagules has led to a more detailed identification of potential health hazards at the working place. In addition to the pathogenic and allergological relevance, airborne fungi are thus of toxicological concern. PMID- 10803223 TI - [Fungal allergen (antigen) burden of refuse disposal workers in biological waste and garbage collection within the scope of a work site analysis]. PMID- 10803224 TI - [MVOC of fungi--use as an indicator for exposure level]. AB - The production of microbial volatile organic compounds (MVOC) by fungi derived from biowaste was studied on synthetic agar (YES), compost-extract media (CEA VAR 1-3) and compost. Exposure assessment was carried out in composting facilities and the occurrence of VOC/MVOC was related to the presence of airborne fungi. A number of non-specific MVOC was found at all sampling sites or in each season, some of which showed preferences for certain sampling sites (3-methyl-1-butanol, 3-octanone). They seemed to be correlated with a certain species composition or with microbial activity, and may thus be used to describe a certain state of decomposition. Correlations between single volatiles and certain fungi were found in situ, but these findings did not match with the species-specific volatiles obtained from pure cultures. PMID- 10803225 TI - [Microbiological emissions from composting sites and their environmental medicine relevance for the neighborhood. Measuring emissions and epidemiological study of 3 Hessian composting sites]. PMID- 10803226 TI - ["Microbial air pollutants--methods for detection of endotoxins, mycotoxins and MVOC"]. AB - In the context of microbial emissions from composting facilities the methods for the detection and identification of the groups of substances released, i.e. endotoxins, mycotoxins and Microbial Volatile Compounds (MVOC) are discussed. With the aid of an overview of the different methods employed for the investigation of the single groups of compounds the current state of the art in this field is presented. In conclusion the enormous research needs, especially with regard to the mycotoxins and MVOC, are pointed out. PMID- 10803227 TI - [Detection of airborne cultivatable microorganisms from compost sites--emissions and imissions]. AB - The statement presented here gives an overview and assessment of the procedures and concepts currently used for the collection and determination of airborne, culturable microorganisms at sources of emission within composting plants and in their near vicinity. The paper focuses on "classical" methods, which involve cultivation as an intermediate step for the determination of viable, airborne microorganisms. The theoretical and practical requirements on such methods are discussed. Results and experiences from recent investigations are described. PMID- 10803228 TI - [Recent methods for the detection of airborne microorganisms and source identification]. AB - The detection of airborne microorganisms including selected cell constituents (e.g. allergens or endotoxins) depends on suitable methods and instruments for their collection. Furthermore, microbiological methods are necessary for their quantification and qualification. In the past these methods were largely based on the classical cultivation dependent approach. Modern molecular methods, e.g. direct staining procedures, hybridization assays with nucleic acids including the PCR-technology or immunological assays are promising new tools for a more sophisticated detection of bioaerosols. They allow a better detection rate, a more precise identification of certain members of the aerosol including cell constituents. With respect to speed and lower costs they are an important alternative to established detection methods. PMID- 10803229 TI - [Effect of microbial aerosols on the human]. AB - Based on numerous publications dealing with the effects of microbial emissions on workers in waste processing plants and on few papers concerning the assessment of the environmental health relevance of microbial aerosols on people living in the vicinity of composting facilities, the current state of the art in this field is presented. With regard to occupational exposure the possible health effects like infections, toxicity and allergies are specified. Since to date only few studies have been made of populations exposed to microbial aerosols in ambient air, the environmental health aspects are reviewed in the context of a study of three compost plants in Hesse, where ambient air measurements as well as epidemiological investigations were carried out. Final recommendations are given and the research needs regarding the environmental health significance of microbial aerosols are formulated. PMID- 10803230 TI - ['Microbial air pollutants'. Emission sources and preventive measures]. AB - Already in the planning or monitoring phase, measures have to be realized for the reduction in emissions of bio-aerosols in biological waste treatment plants. For this purpose, operation-related measures (in open and partially closed plants) and technology-related measures (in closed plants with automatic ventilation) can be taken. A survey on the currently feasible measures for reduction is submitted. The realization has to be adapted to the individual site and type of plant. An enormous demand for examinations is envisaged in order to enable a qualified assessment of the emission behaviour of diverse waste air systems or process steps. PMID- 10803231 TI - [Microbial emissions in collection of residential garbage]. AB - Two exemplary investigations with different questions were carried out in Munich and Stuttgart by sampling bio-aerosols due to and during the collection and emptying of different types of waste bins. The emissions were regarded qualitatively and quantitatively, in order to answer the question, whether and to which amount differences of bio-aerosol emissions occurred due to the kind of wastes collected. The lowest emissions of airborne microorganisms were found during the collection of the paper waste fraction. In comparison, the amounts of them during the collection of unseparated household wastes as well as source separated bio-wastes and the remaining residual waste fraction were higher. Between the three latter, no significant differences could be found, only a tendency of higher values for the thermotolerant mould A. fumigatus during the collection of bio- and residual wastes was observed. Maybe this was caused by special features of the different sampling locations, but it could also be due to the longer collection intervals (bio- and residual wastes were partially collected every two weeks, unseparated wastes every week). PMID- 10803232 TI - Bioaerosol exposure and related health effects in waste collection. PMID- 10803233 TI - [Microbial exposure in collection of residential garbage--results of field studies]. AB - Since 1995 the communal accident insurance carrier of the county Wetfalen-Lippe conducts investigations into the exposure to biological agents related to refuse collection. Total fungal exposure during refuse collection turned out to range from 10,000 up to 750,000 colony forming units per cubic meter. Most of the measurement values exceeded the limit of 50,000. During hot periods in the summertime, the concentration of Aspergillus fumigatus increased up to 90,000 cfu/m3. The mean values of the bacterial concentrations ranged from 15,000 up to 50,000 cfu/m3, the endotoxin concentration from 12 up to 59 EU/m3. In the driver's cabin fungal exposure sometimes exceeded 10,000 cfu/m3 especially in autumn and winter. Maximum values were 5,000 cfu/m3 for bacteria and 15 EU/m3 for endotoxins. High values were measured irrespective of the kind of refuse. PMID- 10803234 TI - Endotoxin and beta-(1-->3)-glucan exposure in household waste collectors and compost workers: a relation with upper airway inflammation. PMID- 10803235 TI - [Risk to employees in garbage collection and transport from pathogen exposure]. AB - In a current study funded by the Federal Institute of Occupational Safety and Hygiene the microbial exposure during waste collection and the health of waste collectors are being examined. The investigation of the exposure is carried out in standard experiments and in field studies. It should give information about the extent of emissions of bacteria and fungi spores und confounding factors. Medical examinations of waste collectors will show if there are special health risks attributable to waste collection. PMID- 10803236 TI - Health hazards to waste management workers in Denmark. AB - This paper deals with the Danish investigations of garbage recycling workers (GRW) in the beginning of the 90ties. The studies are summarized here showing a wide range of symptoms and diseases, from occupational asthma to gastrointestinal and skin symptoms. From the Danish experiences it seems that under normal circumstances with a good hygiene and use of the proper protective equipment by an educated work force, garbage handling induces a small but significant risk of occupational asthma. The majority of the asthma cases we have experienced in Denmark have been due to a poor perception of the risks related to organic dust exposure. However, there is still an increased prevalence of respiratory symptoms such as chest tightness and toxic alveolitis. Furthermore gastrointestinal and skin symptoms are more frequent among GRW compared to other blue-collar workers. PMID- 10803237 TI - [Waste management--health: a longitudinal study]. AB - Altogether 256 (148 men and 108 women) employees from manual sorting facilities and 120 (98 men and 22 women) employees from compost facilities were examined The main focus of the investigations was in the assessment of the lung-function and the immune-system. A part of employees (87 in sorting and 34 in compost facilities) was observed over a period of 3 years. The lung-function was in the area of the expected values, decreased however from year to year. The concentration of total-IgE increased over the period of 3 years. The observation is from great interest that the difference of the concentration of the total-IgE fluctuated within the 3 years at the employees of manual sorting facilities from +200% to -100% while it amounted to 0% at employees of the compost facilities. PMID- 10803238 TI - [Occupational protection in biological waste treatment plants from the occupational medicine viewpoint]. AB - Composting of biogenous waste matter constitutes an important segment in the Federal Republic of Germany's developing waste and recycling management. The processes used in compost production entail exposure to bio-aerosols, gases, dust, microorganisms and their toxins, additionally noise, fluctuating climatic conditions and various accident dangers. A study carried out was task with elaborating a specific profile of strains on compost workers in comparison with a control group. 42 composting plants were visited in 1996 and 1997. In the plants, spread out over seven federal states, 184 compost workers participated in 1996 and 178 in 1997. The control group comprised 66 (1996) and 63 (1997) employees of a senior state authority with office and laboratory jobs. About 400 mainly humanly relevant air measurements for meso-phile/thermo-phile actinomycetins or mould fungi provided emissions data for the purpose of estimating biological risks. Jeopardy analysis of jobs for chemical and physical jeopardy factors was based on the checklist method supported by GC-MS screening as well as measurements of noise level, light density and dust concentrations. On-site medical examination of all 491 employees comprised detailed industrial-medical anamnesis, internistic-clinical physical examination, recording of static-dynamic pulmonary function parameters as well as haematological-immuno-serological screening. From the results, it can be concluded that ailments of the respiratory tract and the lungs as well as chronic bronchitis, bronchial asthma and exogenous allergic alveolitis are not found with significantly greater frequency in compost workers than they were in the control group. The same is likewise true for allergies, diseases of the locomotor system and the skin. ODTS did not occur. During the study period, there was one case of occupational disease in a compost worker with an hypersensitivity pneumonitis caused by Aspergillus fumigatus. Efforts are necessary for a better filter technique and the reduction of noise and carbon dioxide hazards. PMID- 10803239 TI - [Health risk to workers in recycling plants and on waste disposal sites]. AB - Two groups of workers in garbage sorting plants and waste landfills and a control group of community employees were physically examined. In garbage sorting plants elevated microbial counts were measured in the air of the workplace compared to outdoor air microbial counts (cfu/m3). Workers in garbage treatment plants were higher exposed to airborne microbial agents. In consequence immunological reactions and the occurrence of workplace related symptoms were found more frequently, These symptoms are described in literature as mucous membrane irritation syndrome (MMI). More infectious diseases were seen in workers in garbage treatment plants than in the control group. As significant more health problems were seen in groups exposed to very high concentrations of microbial agents in the air at the workplace (moulds more than 10(5) cfu/m3, bacteria more than 10(4) cfu/m3), this study supports the recommendations in the LASI guidelines 1, 13 and 15 and the TRGS 540 to reduce the microbial load in the air at the workplace. PMID- 10803240 TI - [Microbial exposure level and emission values of biological aerosols on refuse disposal surfaces composed of residential garbage and industrial waste]. PMID- 10803241 TI - [Measuring the spread of airborne microorganisms exemplified by 5 biological waste management sites]. PMID- 10803242 TI - [Measuring the spread of airborne microorganisms in the area of composting sites]. AB - Emission concentrations of culturable microorganisms were determined in the vicinity of three open or partly encapsulated composting facilities. Sampling was performed during so-called worst case situations which should promote aerial transport of emissions. Suitability of thermophilic organisms to detect an emitting influence of the plant was confirmed. Generally, concentrations decreased significantly with increasing distances from the plant at all three locations. At one plant 10(6) CFU m-3 thermophilic actinomycetes were found in a distance of 200 m. Partly increased concentrations could be determined even in distances above 500 m. Concentrations could vary within one hour to more than ten times. PMID- 10803243 TI - [Optimizing technical and microbiological procedures in biological waste air management]. PMID- 10803244 TI - [Determining the emission of microorganisms from biofilters and emission concentrations at the site of composting areas]. AB - In order to assess source emissions and dispersal of airborne culturable microorganisms from composting plants, measurements at three composting plants have been carried out. The downwind concentrations of dispersed microorganisms differed greatly, depending on the type of plant design. At 200 metres downwind from the totally enclosed composting plant, levels of spore concentrations of thermotolerant fungi and Aspergillus fumigatus, which may be regarded as characteristic for composting operations, were not above the magnitude of background concentrations. In contrast, spore concentrations in excess of the background level occurred within 500 metres of the partly open plant. Moreover, the ranges of airborne concentrations at similar distances from the enclosed plant were much smaller relative to the partly open plant. Measurement of source emissions from biofilters showed concentrations in the raw and purified gases in the range of up to three orders of magnitude. The operational characteristics of the plants generally contributing to bioaerosol emissions on-site were found to have an influence on the concentration levels in the raw gas. A decrease in the microbiological parameters which may be regarded as specific for composting operations, was attributed to a reduced rate of passage through the biofilters. The magnitude of reduction as well as the concentrations varied greatly. PMID- 10803245 TI - [Comparative studies of airborne, cultivatable microorganisms at selected sites of waste managements, domestic animal husbandry and in the surrounding residential area]. AB - During the last years, sampling of airborne microorganisms (including mesophilic bacteria, moulds, thermophilic bacteria and A. fumigatus) in waste disposal, recycling industry and control sampling locations in the urban and rural districts of Graz was conducted using Andersen 6-stage viable cascade impactors. In the present study additional sampling in livestock buildings has been conducted. Except mesophilic bacteria the emission in the area of waste disposal and recycling industry was significantly higher than in livestock buildings. In the surroundings of the livestock buildings the number of microorganisms was not increased, while the normal background level in the surroundings of the waste proceeding plants was reached from a distance of 150 to 300 meters. PMID- 10803246 TI - [Composting with semipermeable covers, a possibility for reducing emissions of microbial aerosols]. PMID- 10803247 TI - [Susceptibility to selected chemotherapeutics of Staphylococcus aureus strains resistant to methycyline isolated from clinical materials in the years 1991-1992 and 1997]. AB - The susceptibility to selected chemotherapeutic agents was determined in 100 strains of Staphylococcus aureus methicillin-resistant (MRSA) isolated from clinical materials in 1991-1992 (50 strains) and in 1997 (50 strains). Two methods were used for the determination: disc method and antibiotic dilution in agar. The minimal inhibitory concentration (MIC) was determined for vancomycin, teicoplanin, furazolidone, nitrofurantoin, ofloxacin, gentamicin, netilmicin and trimethoprim. The concentrations of the chemotherapeutics in the substrate ranged from 0.125 to 512 mg/l. The obtained results served for drawing of the following conclusions: all studied MRSA strains isolated in 1991-1992 and in 1997 were sensitive to glycopeptide antibiotics: vancomycin and teicoplanin, to nitrofurans: nitrofurantoin and furazolidone, and to fusidic acid. MRSA strains isolated in 1991-1992 were sensitive to ofloxacin, but in 1997 about 80% of the strains were resistant to that antibiotic, and this resistance was noted in S. aureus strains with homogeneous resistance to methicillin. Increasing frequency of resistance to mupirocin was found, in 1991-1992 4% of the strains were resistant, and in 1997 the resistance of MRSA to that antibiotic was found in 12%. No changes occurred in the sensitivity of staphylococci to trimethoprim/sulfamethoxazole (cotrimoxazole). About 94% of strains in 1991-1992 and 1997 were sensitive to that drug. The sensitivity to cotrimoxazole is connected with one of its components (trimethoprim), with 94% of MRSA strains sensitive to it. PMID- 10803248 TI - [Genetic transfer of methycilline resistance in Staphylococus epidermidis and Staphylococcus aureus strains in mixed cultures]. AB - In mixed cultures of staphylococci a transfer of the resistance to methicillin and penicillinase plasmids as well as tetracycline and chloramphenicol plasmids was investigated. It was shown that the resistance to methicillin was transferred in mixed cultures from one strain of S. aureus to another and from S. epidermidis to S. aureus. In both cases transfer of methicillin resistance required, the presence of penicillinase plasmid in recipient or donor strain. In the case of other markers transmission was independent. Moreover it was shown that the transfer of resistance genes in mixed cultures was mediated by bacteriophage of the serologic group A. PMID- 10803249 TI - [Use of pulse-field electrophoresis for intraspecies differentiation of methycillin-resistant, coagulase-negative strains of Staphylococcus aureus (MRSA CN)]. AB - Strains showing a negative reaction in tube test for coagulase accounted 10 to 20% of all Staphylococcus aureus isolated from patients of PSK No. 1 in Warsaw. They are MRSA as well as MSSA strains. 37 coagulase-negative isolates of MRSA were examined using the method depending on digestion of whole cell DNA with SmaI enzyme and electrophoretic separation of the obtained fragments in the pulsed field (PFGE). It was shown that majority of the strains (26 from 37) had individual, unique patterns of bands. However, the two groups of strains were also observed showing a great similarity. The larger group contained 8 strains which were obtained from patients from different wards and the smaller group contained 3 strains obtained from patients from one hospital ward. The obtained results showed that among coagulase-negative MRSA strains are not only sporadic strains but also strains with probably epidemic properties, derived from different clones. PMID- 10803250 TI - [Typing of methicillin resistant strains of S. haemolyticus isolated from patients and from the hospital ward environment]. AB - Biotyping, antibiograms and fingerprinting were used to determine the relation of 16 methicillin-resistant S. haemolyticus isolated from drains in patients who underwent intraabdominal surgery to 9 methicillin-resistant strains of S. haemolyticus isolated at the same time from hospital environment. The comparison of biochemical properties of the examined strains showed a large variety of biochemical profiles as well as antibiotic patterns of susceptibility. The differences in sensitivity to the antibiotics used were not distinct. Biotyping and antibiograms did not permit determination of the relation of the investigated strains. Only the results of fingerprinting allowed for the division of the 25 examined strains into three genotypes demonstrating three main patterns of PCR products. 16 strains of 25 showed the same pattern of PCR products. This results suggests the presence of a source of infection on the clinical ward. A nurse may have been the source of infection because the same genotype of S. haemolyticus was isolated from her nasal anterior as from the majority of patients. PMID- 10803251 TI - [Studies on siderophore exchange properties between staphylococci and various species of gram-positive and gram-negative bacteria]. AB - The ability of iron utilizing by means of staphylococcal siderophores by bacteria belonging to genera: Acinetobacter, Corynebacterium, Curtobacterium, Clavibacter, Bacillus and Mycobacterium was investigated. The staphylococcal donor strains (18 species) used in these experiments were characterized by the ability to utilize siderophores produced by various strains belonging to aforenamed genera. The utilization of staphylococcal siderophores was studied on agar media in which minimally effective concentrations of ethylenediaminedi-ortho-hydroxyphenylacetic acid (EDDA) were used to inhibit indicator strains. Test colonies (staphylococcal) were applied to the surface of the media to determine whether the indicator organisms could obtain the required iron for growth by utilizing chelators from the test colony. The growth inhibition by EDDA of most strains from the Acinetobacter rods and from the coryneform-organisms (plant pathogen) genera, and strains from the species: B. subtilis, M. phlei, M. smegmatis, M. fortuitum was reversed by staphylococcal siderophores. None of the staphylococcal strains investigated, had the ability to exchange siderophores with strains from the species: C. pseudodiphtheriticum, Corynebacterium ANF group, B. megaterium, M. vaccae, M. chitae and M. parafortuitum. PMID- 10803252 TI - [Drug resistance of Enterococcus species isolated from the urogenital system]. AB - Despite low virulence of enterococci, they have become important nosocomial pathogens. This has been correlated with the increased use of broad-spectrum antibiotics, particularly cephalosporins. Many strains of enterococci exhibit multiple drug resistance; the most important being high-level resistance (HLR) to penicillin (MIC > 100 mg/l) and gentamicin (MIC > 500 mg/l and 2000 mg/l) and/or streptomycin (MIC > 2000 mg/l). The investigation was performed on 92 strains, isolated from genito-urinary tract and recognised as Enterococcus sp. All strains were obtained from several microbiological laboratories of Gdansk, Gdynia and Tczew. On biochemical reaction profiles species of enterococci were identified as: E. faecalis (72.8%), E. faecalis varians (9.8%), E. durans (7.6%) and E. faecium (9.8%). The minimal inhibitory concentration (MICs) of penicillin, ampicillin, azlocillin, imipenem, gentamicin, amicacin, ciprofioxacin and vancomycin were determined by the agar dilution method. None of these 92 enterococcal strains was vancomycin resistant. 22.2% of E. faecium and 7.5% of E. faecalis showed high-level resistance to penicillin. None of these strains were produced beta-lactamase. High-level resistance to streptomycin and gentamicin was detected. Both--high-level resistance to streptomycin and gentamicin--were found in 6% E. faecalis; 11.1% E. faecalis varians and 22.2% E. faecium. PMID- 10803253 TI - [Drug resistance of 100 clinical strains of Enterococcus spp]. AB - The aim of this study was to evaluate the drug susceptibility of 100 Enterococcus spp. strains isolated from patients hospitalized in State Clinical Hospital No 1 in Warsaw. All strains were identified (API 20 STREP) and their susceptibility to antibiotics was tested (ATB STREP) in automatic ATB system. Additionally, PYRase activity, beta-lactamase production (in nitrocefin test), MICs for vancomycin and teicoplanin (E test), HLAR--high level aminoglycoside resistance and susceptibility to vancomycin, teicoplanin, piperacillin and piperacillin/tazobactam (disc diffusion method) were determined. E. faecalis ATCC 29212 was used as the control strain. Fifty E. faecalis, 45 E. faecium, 2 E. casseliflavus, 2 E. durans and 1 E. avium strain were cultured. All strains were PYRase-positive and beta-lactamase-negative. Ten isolates demonstrated intermediate susceptibility to vancomycin (6--E. faecalis and 4--E. faecium). One E. faecalis strain was intermediately susceptible to both glycopeptides. One E. casseliflavus strain showed low-level resistance to vancomycin, but this strain was susceptible to teicoplanin--phenotype Van C. HLAR strains were found among 31 E. faecalis and 40 E. faecium strains. 48 E. faecalis strains were susceptible to piperacillin and 49 to piperacillin/tazobactam. Whereas, 41 E. faecium were resistant to both these drugs. Thirty six per cent of isolates were resistant to penicillin and ampicillin, 73% to erythromycin, 87% to tetracycline, 89% to lincomycin and 56% to nitrofurantoin. Some discrepancies were noticed between the results of different methods applied for susceptibility testing--ATB system, E test and disc diffusion. These discrepancies concerned HLAR detection and susceptibility to glycopeptides determination. The best methods were: disc diffusion for HLAR detection and E test for determination of resistance to vancomycin and teicoplanin. Increasing resistance to antimicrobial agents is observed in clinical Enterococcus spp. isolates cultured in our laboratory, especially in E. faecium strains. It is necessary to control the dissemination of multiresistant Enterococcus spp. strains in hospital wards. PMID- 10803254 TI - [Hydroxamate siderophores in enterococci]. AB - In 70 enterococcal strains of diverse origin belonging to 16 species a hydrooxamate class siderophore was detected with chemical and biological tests. A correlation between hydrooxamate siderophore production, species affilIation and pathogenicity of enterococci was not found. PMID- 10803255 TI - [Humoral reaction to Bordetella pertussis antigens: pertussis toxin, filamentous hemagglutinin and lipopolysaccharide in children with clinical symptoms of whooping cough. I. Antibody level for B pertussis antigens in children without respiratory tract infection symptoms]. AB - The aim of the first part of this study was to determine antibody level to pertussis toxin, filamentous hemagglutinin and endotoxin of B. pertussis in children without symptoms of respiratory tract infection. The serum samples obtained from 276 children (age range: 6 weeks-16 years) were examined using indirect hemagglutination and ELISA tests. Normal antibody levels to 3 B. pertussis antigens were determined for 95% of the serum samples as the upper cut off levels depending on children age. Very high level of IgG antibodies to B.perussis antigens was observed in the control population. The lowest antibody level was found in IgA class to pertussis toxin and lipopolysaccharide. It was also established that the IgM level to 3 B. pertussis antigens was rising together with children age. PMID- 10803256 TI - [Humoral reaction to Bordetella pertussis antigens: pertussis toxin, filamentous hemagglutinin and lipopolysaccharide in children with clinical symptoms of whooping cough. II. Occurence and level of B. pertussis antigens in children with suspected whooping cough]. AB - The aim of this study was to determine and evaluate IgG, IgM and IgA levels to pertussis toxin (PT), filamentous hemagglutinin (FHA) and endotoxin (LPS) of B. pertussis in children with clinical symptoms of whooping cough. The serum samples obtained from 265 children (age range: 2 months-16 years) suspected of pertussis were examined by indirect haemagglutination (IH) and ELISA tests. Higher antibody level was most frequently observed in IgA class to PT, FHA and LPS in 45.3%, 35.1% and 66% of pertussis patients sera respectively. The least positive results were obtained in IgM class to PT and FHA (in 9.8% and 2.6% of children sera respectively) but in the case of LPS applied as the antigen in ELISA, higher IgM level was determined in 46.8% of pertussis patients sera. The four times increase of antibody level to LPS determined by IH was observed in 86.7% of children suspected of pertussis. Humoral response to B. pertussis infection is mainly connected with higher IgA level to PT, FHA, LPS and IgM to LPS in children with clinical symptoms of whooping cough. PMID- 10803257 TI - [Use of WARD et al's scheme for bacteriophage typing of Salmonella enteritidis strains isolated in Poland]. AB - The typing phages set of Ward et al. was used to type a total of 517 strains of Salmonella Enteritidis isolated in Poland in 1986-1995. According to the Ward et al. scheme, 56.5% of the strains tested were assigned to 14 different phage types. Phage types 8, 4, 1 and 4a were placed first, second, third and fourth, respectively. They were dominated both in the outbreak isolates and in the isolates from the other sources. Ten phage types were represented by single strains. Other strains reacted with phages without showing any of the designated phage type (37.1%) or were untypable (6.4%). The Ward et al. scheme seems to demonstrate not enough high degree of strain discrimination for Salmonella Enteritidis isolates in Poland. It seems that the Ward et al. scheme is not enough useful for the epidemiological investigations of Salmonella Enteritidis in Poland. PMID- 10803258 TI - [Taxonomy of Citrobacter rods found in human specimens]. AB - The aim of this study was the identification of 181 Citrobacter strains on the basis of the recently proposed taxonomic changes of Brenner. All strains were isolated from diarrhoeic patients; 124 strains were originally sent for identification to Laboratory of Enterobacteriaceae DB NIH, 57 strains was isolated in Czech Republic. Citrobacter isolates were initially identified as C. koseri (3 strains), C. amalonaticus (1 strain) or as members of the C. freundii complex (177 strains). Additionally some biochemical tests were performed. The ability to grow in medium containing KCN, lysine decarboxylase production, lactose fermentation and PYR test were examined. Strains belonging to the C. freundii complex were identified to the species level by biochemical methods on the basis of the results of Brenner, who found some tests to be useful in separating Citrobacter species. These test included citrate and acetate utilization, arginine dihydrolase and ornithine decarboxylase activities, motility, urease production, esculin hydrolysis, and acid production from sucrose, dulcitol, melibiose, raffinose and salicin. On the basis of the criteria described above, 96.6% of the strains tested could be assigned to one of the recently named species of C. freundii complex. Using biochemical tests suggested by Brenner we were able to identify Citrobacter strains members of newly recognised species. A five-test system is proposed to identify the most frequently encountered species currently residing in the C. freundii complex. PMID- 10803259 TI - [Differentiation of Citrobacter strains using electrophoretic protein patterns]. AB - The aim of this study was checking of the usefulness of electrophoretic protein patterns in differentiation of Citrobacter strains. For analysis of whole-cell proteins 181 Citrobacter strains were prepared. Electrophoresis was performed in Mini Protean Duall Slab Cell (Bio-Rad) apparatus. Electrophoresis was carried out in 10% polyacrylamide gel according to the SDS-PAGE method of Laemmli. Whole-cell proteins of all tested Citrobacter strains gave after electrophoresis 12 to 20 bands. Patterns consisting of 12 to 20 fragments ranging in size from 70,000 to 14,000 and smaller, were not distinguishable. There were no significant differences between electrophorograms of Citrobacter strains belonging to the different species, useful for species differentiation. Identical protein band patterns were observed in the case of selected strains e.g. strains C. sedlakii studied in this study, coming from an outbreak, having the some phenotype. PMID- 10803260 TI - [Differentiation of Citrobacter strains using chromosomal DNA restriction patterns]. AB - The aim of this study was checking of the usefulness of chromosomal DNA restriction patterns in differentiation of Citrobacter strains. Molecular characterization of total 56 isolates of Citrobacter from Poland and Czech Republic, was performed by pulsed-field gel electrophoresis after digestion of chromosomal DNA with restriction endonuclease Xba I (5'-TCTAGA-3'). Chromosomal DNA of all tested Citrobacter strains gave after electrophoresis 12 to 21 bands and patterns consisting of 12 to 21 fragments ranging in size from 790 kb to 48.5 kb and smaller, which where not distinguishable. Pulsed-field gel electrophoresis patterns were useful for comparing Citrobacter strains. Identical restriction patterns generated by PFGE were observed in the case of selected strains e.g. strains C. sedlakii studied in this study, coming from an outbreak, having the some phenotype. In addition, PFGE patterns can be used to evaluate the clonal relatedness among bacterial isolates. PFGE can be helpful for assessing genetic relatedness among strains epidemiologicaly unrelated e.g. C. werkmanii strains tested in this study. The sum of DNA fragments after Xba I digestion indicates the genome size of Citrobacter strains. This suggests that PFGE should be useful for epidemiological investigations of Citrobacter strains. PMID- 10803261 TI - [Drug resistance and proticinogenic types of Proteus mirabilis isolated from urinary tract infections]. AB - Proteus mirabilis strains (88 isolates) from hospitalised patients with urinary tract infection were tested for antibiotic susceptibility, ESBL production and their ability to produce proticin or on their susceptibility to proticin. Antibiotic susceptibility test was performed by standard disc diffusion method according to NCCLS. Proticin typing was made by the standard strain set from the B. W. Senior collection. Most (59%) strains belonged to ESBL producers and were more resistant to antibiotics than ESBL-negative strains. Predominant proticin patterns among the ESBL (+) strains were: P1,2(6)/SO (23%), P1,2/SO (13.5%). Among the ESBL-negative strains more frequent were P6/SO (16.6%), P1,2/SO (13.8%) and P3,6/SO (13.8%) proticin types. PMID- 10803262 TI - [Susceptibility to selected antibiotics of Yersinia enterocolitica 03 strains, carrying and not carrying plasmid pYV]. AB - A total of 199 clinical strains of Yersinia enterocolitica serotype O3, biotype 4 were tested for their susceptibility to antibiotics (158 strains carried the virulence plasmid pYV and 41 strains did not). 114 isolates were tested by standard disk diffusion method for 21 antibiotics. Almost all tested strains were resistant to ampicillin and cefazolin and susceptible to amoxycillin/clavulanate, cefaclor, cefamandole, cefuroxime, cefotaxime, ceftriaxone, aztreonam, imipenem, gentamicin, amikacin, netilmicin, tetracycline, doxycycline, chloramphenicol, ciprofloxacin, sulphamethoxazole, co-trimoxazole, trimethoprim and furazolidone. In addition minimal inhibitory concentrations (MICs) of 15 antibiotics were determined by agar dilution method for all 199 strains (158 plasmid positive and 41 strains plasmid negative). Third-generation cephalosporins such as cefotaxime and ceftriaxone and a fluoroquinolone (ciprofloxacin) were the most active antimicrobial agents, tested followed by aztreonam, imipenem, trimethoprim, tetracycline, gentamicin, chloramphenicol, amoxycillin/clavulanate, cefaclor, cefuroxime, amikacin, furazolidone and sulphamethoxazole. The present study demonstrated a high susceptibility of clinical strains of Y. enterocolitica to most of the tested antibiotics. In general, there was no significant difference between susceptibility of virulence plasmid pYV positive and virulence plasmid negative strains to antibacterial agents. PMID- 10803263 TI - [Bactericidal activity of human, swine and cattle serum against pseudomonas aeruginosa strains]. AB - The aim of this study was to assess of bactericidal activity of human, swine and cattle serum against 136 of Pseudomonas aeruginosa strains isolated from people, fishes, domestic and fur animals. The mechanism of the bactericidal activity of serum against gram-negative bacteria is complex and involves the participation of complement, antibodies and lysozyme (1, 5, 7, 8, 10, 11, 13, 14, 24, 25, 27, 30). The susceptibility of gram-negative rods to serum is differentiated. Pseudomonas aeruginosa strains are the most resistant (17, 25, 30). This opportunistic pathogen produce proteases that destroy complement components and immunoglobulins (3, 18, 19). The bactericidal activity of serum was determined after 3 hours incubation of bacteria in 50% serum by the method of Jankowski (1981) (5). The results of this study indicate that 71% of this strains were resistant to swine serum action, 68% of this strains were resistant to bovine serum and 57% of the Pseudomonas aeruginosa strains were sensitive to human serum. The P. aeruginosa strains isolated from fishes were the most sensitive to serum action and the strains isolated from people and cattle were most resistant to the bactericidal activity of serum. PMID- 10803264 TI - [Bacterial infections of skin and soft tissues in filariasis]. AB - Adenolymphangitis is a common occurrence in filarial lymphedema. Damage to the lymphatics and lymph nodes by F. bancrofti is followed by obliteration of lymph vessels and lymph stasis. Obstruction of lymphatics prevents the bacteria penetrating skin to be evacuated with lymph stream to regional lymph nodes. Colonization of dermis, subcutis and lymphatics evokes clinical symptoms of adenolymphangitis. The question arises which strains of bacteria are responsible for the acute and chronic types of adenolymphangitis. The most probable strains responsible for this condition belong to the cocci and probably the bacillus strains. PMID- 10803265 TI - [Peritonitis determined by the site of intra-abdominal surgery]. AB - The aim of this study was to determine bacterial flora infecting the peritoneal cavity during intraabdominal surgery by site of operation. Three groups of patients were examined. 29 patients who underwent surgery on the stomach, duodenum, biliary tract or pancreas, 15 patients operated on because of acute appendicitis and 63 patients operated on because of colon or rectum tumours. At the end of the operation but before closure cultures were obtained by swab from the completed anastomosis site. Samples were placed into transport medium and transported promptly to the laboratory. The results of the bacteriological examinations showed that the peritoneal cavity of all patients operated on were infected with bacteria characteristic for the digestive tract, especially by Enterobacteriaceae spp., Enterococcus spp. and Bacteroides spp. From patients operated on because of rectum or colon tumours 3 or 4 bacterial species were isolated most often and they were often infected with P. aeruginosa and C. albicans. This was in contrast to patients from the other groups. In patients infected with polymicrobial flora, B. fragilis and E. coli or enterococci and E. coli and enterococci were most often seen. PMID- 10803266 TI - [Microbial analysis of clinical material taken from patients at the Oncology Center, Maria Sklodowkda-Curie Institute in Warsaw in 1997]. AB - An analysis was carried out of the microbiological investigations of clinical material samples obtained from the patients of two oncology centres belonging to the Warsaw Oncology Centre. Microorganisms cultured from urine, blood, catheters, smears of wounds and other materials were analysed. From 4839 clinical material samples from the Ursynow centre 1755 bacterial strains were isolated. From 423 samples from the centre in Wawelska Street 171 strains were obtained. In infections of patients from the centres the number of Gram-positive cocci was twice that of Gram-negative rods. In the investigated clinical material S. aureus was the most frequently isolated Gram-positive coccus, while E. coli was the most frequent species among Gram-negative bacteria. In the infections of oncological patients a considerable frequency was noted of yeast-like fungi, especially C. albicans. Particularly disquieting was the increasing number of isolates of C. glabrata and C. krusei strains resistant to fluconazole. PMID- 10803267 TI - [Use of RT-PCR and nested PCR for diagnosing enterovirus infections]. AB - An application of reverse transcription (RT-PCR) and nested-PCR for the detection of human enteroviruses isolated in Poland is described. A specific 297-bp fragment was amplified by the method from all studied enteroviruses strains. Amplification system and the nested primers allowed to detect RNA isolated from 10(1) TCID50/ml which corresponded to one molecule of viral RNA in tested volume. However, applicability of the method for routine diagnosis of enterovirus neuro infection should be reassessed in extensive clinical studies. PMID- 10803268 TI - [Papillomaviruses and herpesviruses as selected risk factors in the etiopathogenesis of cervix neck cancer. I. Use of nucleic acid hybridization for diagnosing HPV infections]. AB - HPV infections are regarded as the main etiological factor responsible for the presence of cytological abnormalities and the primary risk factor for cervical cancer development. Diagnostic materials were collected between 1995 and 1997 in the Gynecologic Cancer Prevention, Cancer Center Institute of Oncology, Memorial Hospital Maria Sklodowska-Curie, Warsaw. The patients were divided into three groups: group C--women suspected for viral infection during clinical or cytological examination and two comparative groups A and B--women invited for routine cytological examination. Cytological smears for nucleic acid hybridisation collected before cytological smear sampling with a dacron swab (groups A and C) or collected after cytological smear sampling with a cervical brush (group B) were used. Cytological and clinical data was also used in the investigation. In 52% of the 236 samples tested by nucleic acid hybridisation HPV DNA was found to be present. DNA from the high/intermediate HPV risk group was also present in 36% of the samples and in 11% of the samples from the low risk HPV group. In 5% of the samples we confirmed the presence of mixed infections from both HPV risk groups. The results obtained from nucleic acid hybridisation with pap smear results were compared. It was observed that HPV infections from the high risk group occurred more frequently in pap 3 and pap 4 test results; HPV infections from the low risk group occurred more frequently in pap 1 and pap 2 results, while mixed infections from both risk groups occurred particularly in pap 4 and pap 5 results. Women in the 35-45 age groups suffered more frequently from infections from the high/intermediate risk group. In the 25-35 and 55-65 age groups HPV infections from the low risk group occurred more frequently. In the comparative groups only 2.6% of the women were infected with HPV. PMID- 10803269 TI - [Papillomaviruses and herpesviruses as selected risk factors in the etiopathogenesis of cervix neck cancer. II. Use of PCR for isolating DNA of human papillomavirus and Herpes simplex]. AB - The aims of the study were to compare polymerase chain reaction PCR with nucleic acid hybridisation HC in the routine diagnosis of HPV infections. Smears collected for PCR were digested for 24 hours using proteinase K. After DNA extraction 174 samples were tested by PCR with human bglobin primers PG04-GH20. The PCR products were separated in 2% agarose gel electrophoresis stained with ethidium bromide. In 80.6% of the samples 256 base pair DNA fragments were observed in the gel in UV light. These samples were tested by PCR with HPV primers MY09-MY11. In 40% of the samples the presence of HPV DNA was confirmed. Next we carried out PCR using a mixture of two pairs of primers bglobin PG04-GH20 and HPV MY09-MY11. DNA for this study was extracted from 24 samples in which the presence of human DNA was not confirmed in the first PCR test and from 7 untested samples. In 21 cases HPV DNA was found to be present in gel electrophoresis. The presence of HPV DNA was confirmed in 44.75% of the samples. PMID- 10803270 TI - [The influence of antibiotics on phagocytic and bacteriocidal activity of rabbit peritoneal macrophages stimulated by filtrates of cultured t-lymphocytes]. AB - The aim of the study was to determine the influence of twelve antibacterial antibiotics (various concentrations) on the activation of rabbit peritoneal macrophages. Macrophages were stimulated by filtrates of culture of lymphocytes T obtained from OVA immunized rabbits. Phagocytic activity and intracellular killing against Listeria monocytogenes were tested by fluorescence method. Penicillin G (0.4-50 mg/l), erythromycin and lincomycin (2.5-40 mg/l) used at all concentrations, were not exerting significant effects on activation of peritoneal macrophages. Cephalosporins, aminoglycosides, and rifampicin at low concentrations (0.4-5.0 mg/l) had no influence on phagocytosis and intracellular killing, also. Cephalosporins at concentration 10 mg/l (cephradine and cefamandole) and 50 mg/l (cefotaxime) inhibited intracellular killing and phagocytic activity. The same results were observed with ampicillin and ticarcillin (50 mg/l). The highest suppression effect was demonstrated using rifampicin at concentration 10 mg/l or more. Gentamicin, streptomycin and amicacin at concentrations 40 mg/l or more significantly inhibited macrophage activation in response to filtrates lymphocytes of culture. These inhibitions were more marked with gentamicin (10 mg/l) than amicacin (20 mg/l) or streptomycin (40 mg/l). All antibiotics did not stimulated the activity of peritoneal macrophages. The suppression activity of peritoneal macrophages by some antibiotics probably acts at the level of specific immune system by interfering with cytokine production. PMID- 10803271 TI - [The effect of selected antibacterial antibiotics on production of interferon gamma (IFN-G) by mouse T lymphocytes stimulated by Listeria monocytogenes]. AB - The aim of the study was to determine the influence of certain antibiotics on the production of IFN-gamma by mouse lymphocytes T after four days incubation with Listeria monocytogenes. The level of mouse IFN-gamma was determined by ELISA method (Inter Test-gamma Mouse IFN-gamma Kit, Genzyme). The strongest immunosuppression effect was demonstrated using rifampicin (39 ng/ml IFN-gamma) (Control: 123 +/- 29 ng/ml IFN-gamma, p < 0.05). Lower immunosuppression effects were observed also with cephradine (54 ng/ml IFN-gamma), amikacin (56 ng/ml IFN gamma) and ticarcillin (83 ng/ml). The obtained results show that all tested cephalosporins (cephamandole, cefotaxime, cephradine) and aminogllycosides (gentamicin, streptomycin, amicacin) inhibit production of IFN-gamma by mouse lymphocytes T. The influence of penicillin G and ampicillin, as well as, erythromycin and lincomycin on the production IFN-gamma was not observed. Our results suggest that rifampicin, ticarcillin, cephalosporins and aminoglycosides act as inhibitors of production IFN-gamma. PMID- 10803272 TI - [Principles of gene therapy]. AB - Treatment of disease by gene transfer into the genome of target host cells has become possible. This article presents current clinical applications of gene therapy and describes possible corrections of genetic defects. PMID- 10803273 TI - [Vectors used in gene therapy]. AB - The ability to direct gene transfer to particular target cells is important to achieve a therapeutic effect. In this review we describe the development of methods for gene delivery to specific cell types. PMID- 10803274 TI - [Relation between local blood flow and bone mineralization indicators in the tibia of female rats]. AB - The results presented contribute to the still unclarified question of the role of bone blood flow in the metabolism and mineralization of the bone tissue. We investigated correlation relationships between 85Sr-microsphere uptake (in % dose per g) or local blood flow (in ml/min per g) and density or ash weight (in g per ml of bone tissue) in the tibia of female rats. In four experiments a total of 16 correlations was computed. The following relationships were found statistically significant (p < 0.05): in exp. A (20 intact females): microsphere uptake--ash weight, blood flow--density, blood flow--ash weight, in exp. B (oophorectomy 4 weeks before the experiment, 28 rats): microsphere uptake--ash weight, blood flow -density, blood flow--ash weight, in exp. C (estradiol benzoate 1 mg s.c. twice a week for 4 weeks, 40 rats): microsphere uptake--density, microsphere uptake--ash weight, blood flow--density, in exp. D (20 oophorectomized females on estradiol): microsphere uptake--ash weight, blood flow--ash weight. The results support the hypothesis of an interconnection between the mineralization of bone tissue and local bone blood flow. PMID- 10803275 TI - Prostaglandin therapy of peripheral occlusive arterial disease. AB - The mechanism of prostaglandin therapy in man is still poorly understood. We therefore investigated in 57 patients Fontaine stage II-IV the effect of a single intravenous infusion (37 POAD patients aged 36-67 years, mean 55.5) and the effect of a series of 14 therapeutic infusions (20 patients aged 32-77 years, mean 60.6). We employed a broad spectrum of methods including Doppler and photoplethysmographic pressure measurements and plethysmographic flow measurements in the toe, foot and calf, I-131 subcutaneous clearance in the lower leg, oral and skin temperature and blood gas analysis. Walking distances were estimated on a treadmill and compared with calf flows and ankle pressures. After a single infusion, blood flow in the foot and particularly in the toe rose while systemic and local pressures remained constant. Foot venous pCO2 dropped and pO2 rose dose-dependently. I-131 clearance did not change. After a series of infusions, ischaemic rest pain subsided and necroses were improved in all subjects with advanced ischaemia except one. In claudicating patients, a remarkable prolongation of walking distances without change in calf blood flow was found. It is concluded that prostaglandin therapy relaxes toe and foot skin vessels and is of considerable value in therapy of advanced skin ischaemia. It may also improve intermittent claudication. The mechanism of action includes, among other factors, a significant drop of local vascular resistance in the ischaemic areas and probably also an increase in capillary fraction of blood flow. PMID- 10803276 TI - [Overview and analysis of publishing 1991-1996 at the the First Medical School of Charles University]. PMID- 10803277 TI - [Man as a subject and object of medicine. A joint seminar given by Charles University and Ruprecht-Charles University in Heidelberg on the occasion of the 650th anniversary of the founding of Charles University, Heidelberg, 14-17 May 1998]. PMID- 10803278 TI - [The beginnings of biophysics worldwide]. PMID- 10803279 TI - [The beginnings of cybernetics and its introduction into the biomedical arena in the Czech Republic]. AB - During fifties at most few tens of persons were in Czechoslovakia who believed in the future of computers and cybernetics. At the sixties, the Czechoslovak Cybernetics was established and in 1962 the Main Problem Committee for the Medical Cybernetics was founded at the Ministry of Health. The first tentative lectures on this topics at the Charles University Medical faculty were introduced in the late sixties. The growing interest enabled to hold national and international conferences since the middle of seventies. There is mentioned the whole spectrum of these goal-directed activities so important for the future. PMID- 10803280 TI - [Modern technical aids in the teaching of medical physics and biophysics]. AB - The set of analogous electric and electronic models of physical and biological processes tested in the teaching process of Medical Physics and Biophysics is discussed. Measuring arrangement of model, digital storage oscilloscope and personal computer allows to display, record, save and process any electric signal. This proposed arrangement offers new possibilities for more detailed description of selected types of signals generally not only in the teaching process. PMID- 10803281 TI - [Tools and methods of developing educational programs in medical biophysics]. AB - In this work various types of programs which can be used in a learning and teaching process are analysed. Also the procedure of creating software for CAL is studied and the procedure, that is the most suitable one for developing CAL is suggested here. Also recommended properties of software for teaching and learning medical biophysics are described. General characteristics and types of authoring tools are discussed in this work and a set of authoring tools is described in more details. Conclusions are documented and demonstrated by program Blood Circulation. For development of this program the authoring system Authorware 3.5 was selected. The program is organized as a hypertext with possibility to find words, with a list of the last visited pages, and with an on line help system. This program works with a text, graphics, interactive animated schemes and with interactive equations. In the program it is possible to test the level of knowledge by means of solving problems or multiply choice test. PMID- 10803282 TI - Glia-axon interactions and the regulation of the extracellular K+ in the peripheral nerve. AB - Changes in membrane potential of both axons and Schwann cells were measured simultaneously during electrical activity and during the period of recovery in the rabbit vagus nerve by the use of the sucrose-gap apparatus. During low frequency stimulation (0.5-1 Hz) the preparation developed a ouabain-sensitive hyperpolarization. This hyperpolarization increased when the inwardly rectifying K+ channels in Schwann cells were blocked with Ba2+, indicating that the hyperpolarization was generated by the electrogenic glial Na(+)-K+ pump. During trains at higher frequencies (15 Hz), the preparation depolarized, but after cessation of the stimulation it developed a posttetanic hyperpolarization (PTH). The PTH was also ouabain-sensitive and was strongly enhanced by Cs+ which is known to block the hyperpolarization-activated inward current (Ih) in axons but not in glial cells. These results show that the PTH reflects mainly the axonal electrogenic pump. Our results indicate that during activity the K+ released from the firing axons is removed from the extracellular space by Schwann cells and that after cessation of the stimulation the K+ surplus returns from Schwann cells back to axons. Both the glial and axonal K+ uptake is mediated by successive activation of the glial and axonal Na(+)-K+ pump. The nature of the signalling mechanisms that control the pumping rates of the respective pumps remain unknown. PMID- 10803283 TI - Changes in mechanical properties of skin as a marker of biological age. AB - The development of the methodology of biological age assessment belongs to the main direction of the research in contemporary gerontology. The sets ("batteries") of tests for biological age assessment involve also large number of typically biophysical measurements. For example measurements of accommodation range, visual acuity, audiometry, vital capacity, blood pressure, pulse frequency and many others. Measurement of skin elasticity is also included in batteries for biological age assessment in many laboratories. As mechanical properties of the skin belong to the quantities strongly modified by aging, they are potential biomarkers of biological age. Unfortunately, the currently applied methods are dubious from metrological and biophysical points of view for many reasons. As a rule, the time of diminishing of some type of deformation is used for this purpose. The subjective errors are present in considerable extent and interpretation of results is also difficult. Above it, this type of measurements is usually called measurement of skin elasticity but, in fact, the process of diminishing of deformation does not depend only on elasticity but depends significantly also on plastic properties of the skin. In our laboratory we suggested and verified methodology which may eliminate the problems discussed above. The methodology is based on theory of viscoelastic bodies. The principle of the methodology is as follows. The human skin (in vivo) is exposed to constant force for limited time. During the same time and after the deformation of the skin is measured. The resulting curve (creep curve) is dependency of deformation on time. The rheological model of the mechanical properties of the skin can be derived from this curve. The model consists of serio-parallel combinations of two elastic (Hooke's) and two plastic (Newton's) bodies. Rheological parameters of these bodies depend on age. The best marker of biological age prove to be modulus of Hooke's body, which is responsible for rapid phase of elastic response. The creep curves were measured in group of 69 persons of both sexes. The age range was between 25 and 82 years. The elastic modulus of Hooke's body responsible for rapid response was calculated in all cases. Further the following measurements was performed in all persons in experimental group: range of accommodation, threshold of hearing of high frequencies of sound (pitch ceiling), vital capacity of lungs, systolic and diastolic pressures of blood (in rest and after exercise), reaction times on visual and acoustic signals. All these markers where evaluated and compared. The following two parameters were taken into account: relative change per one year and coefficient of correlation. The elastic modulus proved to be marker of biological age of the similar quality as accommodation range, pitch ceiling and vital capacity and better than blood pressures and reaction times. The suggested methodology of measurement of mechanical properties of skin seems to be new, well defined biophysical marker of biological age. PMID- 10803284 TI - [Adsorption of derivatives of nucleic acid components on electrodes]. AB - The main advantage of the use of electrochemical methods is that the object studied is under conditions very close to the natural ones. Next advantage is that the same information about the sample can be received with lower costs than other methods. The electrode double layer impedance measurement is effective electrochemical method of the study of interaction of the substances on electrode surface. It is much more sensitive on adsorption processes than other electrochemical methods. After the mathematical evaluation of the results of the electrode double layer impedance measurements it was found, that the adsorption energies of xanthine are much higher than those of adenine and for hypoxanthine are close to adenine. This result could be connected with the role of these two adenine metabolites in nucleic acids. The measurements in the solutions, containing different methyl derivatives of xanthine showed that the adsorption behaviour of every substance is strongly dependent on the position of the methyl group in the molecule. From the dependence of the electrode double layer impedance on the potential and frequency in the solution containing 3'-azido-2'3' dideoxythymidine (AZT) it was found, that under conditions studied a reduction process occurs in the sample near-IV vs. AgCl. The adsorption behaviour of AZT is the same as that of thymidine both at room temperature and at lower ones (near 0 degree C). The measurement of the electrode double layer impedance is the sensitive method for the study of the properties of the nucleic acids components that could be used in the field of new antiviral agents development. PMID- 10803285 TI - [Fluorescence methods for studying cell damage]. AB - We have been studying some properties of the new generation photosensitizers- phthalocyanines. The influence of the certain phthalocyanine concentrations at combination with laser irradiation doses (semiconductor laser, lambda = 670 nm, P = 50 mW) have been studied by in vitro fluorescent methods. Cytotoxicity assay LIVE/DEAD for fluorescence microscopy and measurements of cells viability with multi-well plate scanner was used. The optimal phototoxic effect on MCF7--mamma carcinoma cells was observed 8 micrograms/ml ZnPcS2, 10 micrograms/ml ClAlPcS2 in combination with laser irradiation 50 J/cm2 and 100 J/cm2 respectively. This concentration and dose killed MCF7 carcinoma cells. PMID- 10803286 TI - [Mapping electrocardiographic fields in heart hypertrophy]. AB - The paper deals with chosen parameters that can be used for heart hypertrophy diagnosis and are obtained from body surface electrocardiographic maps. Parameters are analysed in repeated measurements of a patient with left ventricular hypertrophy that regressed due to treatment with verapamil. PMID- 10803287 TI - [Results of measurement of resolution capability of ultrasonographic equipment using our new method]. AB - The quality of sonographic instruments is vital for accurate medical diagnosis. To overcome one subjective problem associated with sonographic equipment, we have designed a system for objective instrument evaluation that is simple, accurate and valid. The system described fulfills the criteria for solving the outlined problem and enables us now to measure the parameters of both the axial and transversal resolutions. Additionally we can appreciate dynamic range of signal, gray scale parameters, the amplitude of video-signal and level of side lobes. To evaluate our method we examined two sonographies with different transducers working on the frequencies of 3 and/or 5 MHz. The method gives reliable results for sonographies quality assessment. PMID- 10803288 TI - [The effect of low-frequency electromagnetic fields on living organisms]. AB - This report studies effect of alternating low-frequency electromagnetic fields (Bm = 5-21.5 mT, f = 50 Hz, duration of exposure t = 0-24 min) on viability of bacteria Escherichia coli. We have shown that the growth of bacteria is impaired the electromagnetic field. Their ability to form colonies on a solid medium decreases in dependence on magnitude of magnetic field and on duration of exposure. The growth curve is influenced by the electromagnetic field as well. Effects of electromagnetic fields are independent of biological age in first four hours of their growth. We have found no morphological changes in bacterial systems in electromagnetic field by optical microscope. Viability of bacteria is bigger in a liquid medium and less in a solid medium. Bacteriophage BF 23 attach less to bacteria influenced by electromagnetic field. And finally, magnetic field did not make induction of production of bacteriophage. This effect indicates, that magnetic field did not damage DNA of exposed bacteria. PMID- 10803289 TI - [The voice and speech]. AB - Biophysics is the science comprising the sum of biophysical disciplines describing living systems. It also includes the biophysics of voice and speech. The latter deals with physiological acoustics, phonetics, phoniatry as well as logopaedics. In connection with the problems of voice and speech, including also their teaching problems, a common language is often being sought for appropriate to all the interested scientific branches. As a result of our efforts aimed at removing the existing barriers we have tried to set up a University Society for the Study of Voice and Speech. One of its first activities was also, besides other events, the realization of a videofilm On voice and speech. PMID- 10803290 TI - [An unknown role of the ligament layer of the conus elasticus during phonation?]. AB - The investigation of the intoning vocal folds with the help of Cermak's mirror in as early as the 19th century and also their ensuring records by means of a stroboscopic film have discovered the fact that the front vibrating part of the vocal folds becomes shortened in high frequencies. This information has been ignored since about the 60ies of the 20th century. The aim of the present paper is to verify the shortening of the front vibrating part of the vocal folds and to recall this fact. Approximately in two octaves we have measured the virtual images of a video stroboscopic record. Our results are comparable with similar measurement resulting from stroboscopic investigation and confirm the hypothetic role of the ligament layer conus elasticus. PMID- 10803291 TI - [A decision-support system for hematology]. AB - Thrombocytopenia and following bleeding at the treatment of hematological malignancies is a dangerous complication. The indication of thrombocyte transfusion is the key point for the therapy and proylaxy of bleeding. The all problem is divided into two parts. Evaluation of the risk of bleeding (80% of decision), estimation of the risk of aloimunization and risk of connected with the transfusion (20% decision). For now we are concentrated to the evaluation of the risk of bleeding. In the first stage we are concentrated to statistical evaluation of values to define factors possibly highering the risk of bleeding. Factors were determined with help of two test, GUHA method and using literature. For recognized factors were trained 3 layer neuron nets with a non-linear method pack propagation. After that an application was developed to determine the risk of bleeding for a routine use in clinical practice. PMID- 10803292 TI - [Computer-assisted decision making in performing transfusions]. PMID- 10803293 TI - [Mortality in congenital abnormalities in relation to age]. AB - The mortality dependence on age after the birth was observed for dominant causes of death in the USA, Japan and former Czechoslovakia for the period 1979-1991. The logarithm of mortality of following causes decrease linearly with logarithm of age at the interval 0-10 years: all, all without accidents, congenital anomalies of heart and circulatory system, spina bifida and hydrocephalus, diseases of the nervous system, other diseases of the respiratory system. The linear area is up to the age of 45 years for Spina bifida and hydrocephalus in the USA. This linear dependence in log-log scale corresponds to the Weibull distribution in case the slope is more than -1. However, the logarithm of mortality of Congenital anomalies decreases in the logarithm of age with the slope of -1. The number of death of congenital anomalies decreases in the logarithm of age with the slope -1, too. The number of death in one year corresponds to the failure density function. The mortality of congenital anomalies is described using another distribution function which is not Weibull function. This new distribution is defined at this paper. The number of death of congenital anomalies in one year is inversely proportional to the age of dead. This phenomenon can be interpreted that the lifetime is determine by the level of malformation at the moment of the birth. The number of people in particular level of congenital anomalies is inversely proportional to the lifetime. PMID- 10803294 TI - Computer assisted evaluation of lipid tests with regard to the risk of atherosclerosis. AB - We developed the algorithm for computer assisted evaluation of lipid tests with regard to the risk of atherosclerosis. This programme has been created as a part of the laboratory information system and is determined to estimate the metabolic risk of coronary artery disease in both screening programmes and lipid clinics. The programme has made an important contribution to diagnostic and therapeutic process of atherosclerosis and hyperlipoproteinemia. It contains evaluation of special indexes, as e.g. HDL-cholesterol/total cholesterol, apolipoprotein A-I/B. PMID- 10803295 TI - [Use of thermoelectric properties of materials with shape memory in medicine]. AB - Article deals with some possibilities of the combination of the shape memory and another physical properties of nitinol (Ti-Ni alloy) for application in the medicine. The thermoelectric properties of originally developed miniature thermocouple probe with the shape memory are described in details. The shape memory effect of the thermocouple is based on the use of the combination of the metals nitinol and constantan. PMID- 10803296 TI - Biophysical inputs into the software "MIRDose". AB - Administered amount of activity decides on absorbed dose in thyroid gland during therapy of thyroid cancer tumors by 131I. Medical Internal Radiation Dose (MIRD) methodology estimates this dose as well as influence on other organs. MIRDose- the software implementation of MIRD--is permanently improving and has reached a substantial degree of maturity. Thus the reliability of the results depends predominantly on quality of the input data. The residence time and functional volume of the thyroid gland of a particular patient are the key inputs. Here we concentrate on the former one. We found that the traditionally used mono exponential model, characterized by the effective half-life, introduces non negligible modelling error. It cannot be improved by any data processing. For this reason, we proposed a novel accumulation model. Now we inspect influences of differences in the guessed residence time on the outputs of MIRDose. We briefly characterize MIRDose software, recall the improved model and present illustrative results of evaluations. PMID- 10803298 TI - Applicability of a circulatory model for determining the parameters of kinetics of endogenously produced compounds. AB - The applicability of circulatory model approach for determining the kinetic parameters of endogenously produced substances has been demonstrated based on the assumption that the behaviour of newly produced molecules of a compound inside its production spaces is the same as that of the molecules returned due to recirculation of blood. While extraction and transmission parameters can be estimated by using tracer data, total mass cannot be determined without additional information. This information could be obtained from data of some suitable metabolite by using double tracer method. Circulatory model is especially suitable for studies of substances with a fast kinetics and renewal if blood (plasma) flow is measured independently. PMID- 10803297 TI - [High pressure technology and examples of its use in the biological sciences]. AB - A short description of basic types of high pressure apparatuses is presented in view of their application in a research in the biosciences. General tendencies in an evolution of characteristic inter-atomic bonds and their changes under high pressure are shortly reviewed. A complex behaviour of organic macromolecular compounds under pressure is demonstrated by the effects of pressure on proteins (including p--T diagram of proteins denaturation). In consequence, an effect of pressure on the simplest micro-organisms is mentioned and relevant critical pressures of sterilization are presented. Trends in a future application of results of the pressure research in biosciences are discussed. PMID- 10803299 TI - Progress in modeling and simulation. AB - For the modeling of systems, the computers are more and more used while the other "media" (including the human intellect) carrying the models are abandoned. For the modeling of knowledges, i.e. of more or less general concepts (possibly used to model systems composed of instances of such concepts), the object-oriented programming is nowadays widely used. For the modeling of processes existing and developing in the time, computer simulation is used, the results of which are often presented by means of animation (graphical pictures moving and changing in time). Unfortunately, the object-oriented programming tools are commonly not designed to be of a great use for simulation while the programming tools for simulation do not enable their users to apply the advantages of the object oriented programming. Nevertheless, there are exclusions enabling to use general concepts represented at a computer, for constructing simulation models and for their easy modification. They are described in the present paper, together with true definitions of modeling, simulation and object-oriented programming (including cases that do not satisfy the definitions but are dangerous to introduce misunderstanding), an outline of their applications and of their further development. In relation to the fact that computing systems are being introduced to be control components into a large spectrum of (technological, social and biological) systems, the attention is oriented to models of systems containing modeling components. PMID- 10803300 TI - [Dendritic crystals as an example of fractal growth]. AB - The crystallization in mixture of physiological solution (0.9% NaCl) with human albumin (model system) or blood serum leads to characteristic dendritic crystals. The composition of blood serum influences crystallization results in dependence on simultaneous pathological process of the organism. The crystallographic method is based on comparison and evaluation of dendritic patterns created by crystallization of NaCl in blood serum. The methods of image analysis are applied to classify the characteristic dendritic crystals in blood serum. This contribution describes possibility of fractal geometry appliance. PMID- 10803301 TI - [Teaching of informatics at the Medical School of Comenius University in Bratislava and its evaluation by students]. AB - In this study we analyzed meanings of students about teaching of informatics at our faculty with aim to improve it. They were asked to complete a questionnaire, which contained questions about organization of teaching, programmes and technical equipment. They had also possibility to suggest their improvements. We sorted the students into two groups according to their previous knowledge and experience in working with computers and then analyzed their answers. The results will be taken into account in further teaching of informatics. PMID- 10803302 TI - [The concept of practical teaching of medical informatics at the Medical School of Palacky University in Olomouc]. AB - The method and conditions of medical information technology teaching represent a direct reflection of both significant progress of information technology and change in public approach to computer techniques. Over the years, the content of the course has been changed to a large extent. It is a result of the existing wide scale of available program products, modernization of local computer network and information technology classrooms and especially common attempt to availability of Internet. The computer-literacy of students is increasing as well as the number and quality of specialized teaching programs and demands of theoretical subjects of study. However the level of computer-literacy of individual students is getting to be widely different. In answer to this situation our institute reacts by some measures, described here. The aim of this paper is to raise a discussion about them and about this issue and to learn more about the approaches and practise in other institutes. In our opinion, a general integration of the main features of the medical information technology teaching courses at Medical Faculties would be the most effective result. PMID- 10803303 TI - [Informatics education from the viewpoint of clinical medicine]. AB - Teaching of medical informatics in the 1st medical faculty is compared with teaching of informatics and related curricula in European faculties. Practically in all faculties in the 1st year, a curriculum is situated with formal (mathematical, probabilistic or information and computer based) approach. From a clinical point of view there are two main reasons for situating this curriculum in the first year--it is a good basis for teaching special parts of informatics later directly in preclinical or clinical curricula and teaching informatics is a good occasion how to bring some clinical problems to the first year study and how to motivate students to the study of theoretical medicine. PMID- 10803304 TI - [Computers in medicine 1998 (conference report)]. PMID- 10803305 TI - Static and dynamic information in metabolic medicine. AB - Several computer models and dynamic parameters were used to describe metabolic state of patients. We have used models to explain the process of glycation of hemoglobin and albumin, to identify insulin sensitivity, to describe individual insulin therapy needs and to predict outcome of metabolic diseases. We also compared static and dynamic description of patients with metabolic X syndrome. We can conclude that static and dynamic parameters bring different quality of information in metabolic care. PMID- 10803306 TI - [The database server for the medical bibliography database at Charles University]. AB - In the medical community, bibliographic databases are widely accepted as a most important source of information both for theoretical and clinical disciplines. To improve access to medical bibliographic databases at Charles University, a database server (ERL by Silver Platter) was set up at the 2nd Faculty of Medicine in Prague. The server, accessible by Internet 24 hours/7 days, hosts now 14 years' MEDLINE and 10 years' EMBASE Paediatrics. Two different strategies are available for connecting to the server: a specialized client program that communicates over the Internet (suitable for professional searching) and a web based access that requires no specialized software (except the WWW browser) on the client side. The server is now offered to academic community to host further databases, possibly subscribed by consortia whose individual members would not subscribe them by themselves. PMID- 10803307 TI - [Providing Internet-based information services at the 2nd Medical School of Charles University]. AB - Development of the internet network at the 2nd Faculty of Medicine, Charles University from one of the first nodes of Prague Academic Network up to a complicated infrastructure with many client computers is accompanied with a corresponding development of information services available in this network. For today's users, infrastructure of the network is hidden behind the services whose technical details are not significant. The paper deals with history, current state and possible perspectives of information services (both basic as e-mail, WWW and more advanced like specialized database server, proxy, etc.) available to users at the 2nd Faculty of Medicine--taking into account the development of information technologies, networking infrastructure and the possibilities and limitations of co-operation between the faculty and its teaching hospital. PMID- 10803308 TI - Clinical informatics in medical education: yesterday, today, tomorrow. Experiences at the first Faculty of Medicine, Charles University. AB - The subject known as clinical informatics started to develop in the 60's in medical education of clinical disciplines and was mainly concerned with structuring information for instruction and examinations. This application contributed substantially to the development of the discipline of medical education that started to be considered as a legitimate new field that could become a clinical sub-specialty in its own right. Due to uneven access to computer hardware the area developed differently in the West and East. We elaborated our projects for application of didactic games in internal medicine for the paper and pencil system. On turn of the 90's the use of PCs brought new possibilities for the application of computer programme support in the analysis of the effectiveness of the system of instruction in internal medicine we were concerned with. We carried out two longitudinal studies in clinical memory of the 4th and 6th year students of the same study cycle. In a non-anonymous questionnaire, more than 1000 respondents described their experience in observation/examination of 70 recommended disorders important for internal practice. We offer some unpublished results showing how the clinical memory of the students is formed during the four-year study and how few possibilities there are for some students to see some manifestations of various clinical disorders. Thinking about the future we are preparing a series of integrated medical and language programmes for the use of the multi-medial approach. PMID- 10803309 TI - [Present status and perspectives in international cooperation in the Cochrane Collaboration]. AB - The Cochrane Collaboration is an international organization that aims to help people make well-informed decisions about healthcare by preparing, maintaining and promoting the accessibility of systematic reviews of the effects of healthcare interventions. It is a non-profit organization. The main work of the Collaboration is done by about fifty Collaborative Review Groups, within which Cochrane Reviews are prepared and maintained. The members of these groups share an interest in generating reliable, up-to-date evidence relevant to the prevention, treatment and rehabilitation of particular health problems or groups of problems. PMID- 10803310 TI - Primary care informatics--the basis of healthcare. PMID- 10803311 TI - [Complex processing of bibliographic records using database applications]. PMID- 10803312 TI - [Regression without derivation and matrices (short report)]. PMID- 10803313 TI - [Financing of education and research in universities]. AB - Universities in the Czech Republic undergo a process of deep changes. There are different perspectives concerning the correctness of the way started with this process. This contribution shows the possibilities which the ministry of Education, Youth and Sports can use to support the quality of teaching as well as the quality of science and research at our universities. The possibilities of evaluation of schools are rather problematic but to find a way of evaluation of the schools is absolutely necessary. There are certain parameters devised in this contribution which will be later elaborated into formulas, then modified in the way which enables that they can be used in the higher education budget allocation in the following year. In the area of research and development there is an evident pressure from the Ministry on the Government to increase the institutional funding of research and development. This funds will be distributed according to the "Rules for the evaluation of research intentions and results of organizations from the view point of the institutional funding provisions for research and development". PMID- 10803314 TI - [In Process Citation] PMID- 10803315 TI - [Vojtech Alexander, the first Slovak radiologist]. PMID- 10803316 TI - Present and future management of prostate cancer. PMID- 10803317 TI - Capromab Pendetide immunoscintigraphy: connecting the dots for prostate cancer imaging. PMID- 10803318 TI - Capromab Pendetide imaging of prostate cancer. AB - Capromab Pendetide imaging illustrates the successful translation of monoclonal antibody technology from the laboratory to the clinic. It provides a means of identifying otherwise occult soft tissue metastases in patients with adenocarcinoma of the prostate. When utilized with other clinical, pathological and laboratory findings, Capromab Pendetide imaging enables more accurate disease staging and monitoring than is afforded by other imaging modalities such as CT and MRI. In the primary disease setting Capromab Pendetide imaging should be reserved for use in patients with negative bone scans who are at high risk for metastatic disease based on such factors as advanced clinical stage, high Gleason score and significantly elevated serum PSA or alkaline phosphatase. Due to low sensitivity for small-volume disease, a negative Mab scan may not eliminate the need for a staging lymph node dissection but should encourage further consideration of local treatment options. Capromab Pendetide should be used with caution in patients at low risk for metastatic disease. Positive scan findings in low risk patients should be confirmed before altering the treatment plan since some false positive scans should be anticipated in a population with low disease prevalence. Capromab Pendetide imaging has not been shown to be reliable in determining the local extent of the primary tumor but new techniques involving co registration of SPECT and CT images show promise in this regard. In the patient with recurrent disease following primary therapy, the predictive value of Capromab Pendetide imaging of the prostate or prostate fossa is limited, particularly following RT. Its more important role in this setting is to identify lymph node metastases in the high risk patient with a negative bone scan who might otherwise be a candidate for local salvage therapy. A large prospective study is needed for confirmation, but preliminary data suggest that Capromab Pendetide imaging is helpful in identifying those patients with PSA elevation after radical prostatectomy who are most likely to benefit from salvage RT. As with any imaging technique, Capromab Pendetide has strengths and weaknesses that must be understood to maximize patient benefit by utilizing the scan in clinical settings where it is most likely to be useful and least likely to be misleading. Capromab Pendetide is a technically demanding procedure best performed and interpreted at sites with experience and expertise. PMID- 10803319 TI - Circumvention of tumor cell escape following specific immunotherapy. PMID- 10803320 TI - Phase I study of CBT-1 and Taxol in patients with Taxol resistant cancers. AB - CBT-1, a natural product, was studied as an MDR modulator with Taxol (135 mg/m2) in an escalating dose Phase I clinical trial. CBT-1 was administered orally at doses from 300 mg/m2 to 500 mg/m2 daily x 7. The MTD was determined to be 500 mg/m2 with moderate nausea and occasional emesis. Side effects were mainly attributable to Taxol rather than the study drug. A total of 18 patients were registered on study with only one patient determined to be intolerant of CBT-1 due to nausea and emesis. In this Phase I study four patients (3 breast, 1 NSCLC) remained stable for greater than two cycles of treatment. No complete or partial responses were seen in this Taxol resistant population of patients with advanced cancer. PMID- 10803321 TI - Short-term tumor cell lines from renal cell carcinoma for use as autologous tumor cell vaccines in the treatment of kidney cancer. AB - OBJECTIVE: We have tried to establish short-term cultures of autologous tumors from patients with renal cell carcinoma that could be used as active specific immunotherapy (i.e., autologous vaccine) in such patients after resection of primary kidney cancer, and/or for the treatment of metastatic cancer. METHODS: Between 10/90 and 9/99 the cell biology laboratory of the Hoag Cancer Center received 69 kidney tumor samples that had been surgically excised, including 43 primary tumors and 26 metastatic lesions. Efforts were made to establish short term tumor cell cultures, as defined by the growth of 10(8) cells; malignant nature and renal cell origin were confirmed by morphology and antigenic phenotyping. Variables associated with successful growth of short-term cell lines were examined. RESULTS: Short-term cell lines were successfully established from 55/69 samples [80%] including 36/43 (84%) from primary tumors and 19/26 (73%) from metastatic lesions. The success rate for tumors harvested at Hoag Hospital was 40/50 (80%); the success rate for tumors obtained from other geographic areas was 15/19 (79%). Tumor cell lines were successfully established from metastatic lesions ranging in size from a 0.5 g vertebral lesion to a 22 g rib/lung chest wall metastasis, and from primary renal cell lesions ranging in size from 1.5 g to 39.7 g. CONCLUSIONS: Short-term cell lines can be established for most patients with primary or metastatic renal cell carcinoma making a pure autologous tumor-cell vaccine approach feasible. Vaccines have been prepared for 41 patients and a vaccine therapy trial is in progress. PMID- 10803322 TI - Establishment and characterization of colon carcinoma and renal cell carcinoma primary cultures. AB - Patients with metastatic renal and colon carcinoma have a very poor prognosis. In many cases, the tumor recurs after surgical excision and chemotherapy. Therefore, it might be beneficial for cancer patients to induce an immune attack against the tumor by inserting a cytokine gene into the tumor cells. Here, two different techniques for isolation of single tumor cells were compared. An enzymatic solution was superior to an EDTA/DTT isolation solution for establishing tumor primary cultures. In total, 18 primary cell cultures could be established from 68 patients with colon and renal cell carcinoma. Cells were further characterized concerning fibroblast contamination, cell proliferation and HLA-typing. These primary tumor cells might be of value for cytokine gene transfer and in vaccination protocols for cancer patients. PMID- 10803323 TI - Chemo-biotherapy with 5-fluorouracil, leucovorin, and alpha interferon in metastatic carcinoma of the colon--a Cancer Biotherapy Research Group [CBRG] phase II study. AB - BACKGROUND: Biochemical modulation of 5-Fluorouracil activity with Leucovorin has been well documented in colorectal cancer. Several studies have shown increased efficacy of 5-fluorouracil in combination with alpha interferon. We therefore initiated a phase II trial of dual modulation of 5-fluorouracil with leucovorin and alpha interferon to evaluate outcomes in patients with metastatic carcinoma of the colon. METHODS: Patients with metastatic colon carcinoma with expected survival > 4 months and performance status of ECOG < or = 2 were treated weekly with Leucovorin 400 mg i.v. followed by 5-FU 600 mg/m2 i.v. bolus. Alpha interferon 3-9 million units was administered subcutaneously every Monday, Wednesday and Friday. Patients were analyzed for toxicity, tumor response and survival. RESULTS: Sixteen patients with a median age of 66 years were treated. Three patients were not evaluable for response but were evaluable for toxicity. Grades 3 and 4 toxicities were neutropenia, diarrhea, mucositis, nausea and vomiting, fatigue, fever, asthenia and elevated hepatic enzymes. One patient died from complications associated with diverticulitis and neutropenia. Objective response rate was 23% (95% confidence interval 4-46%) and median survival was 11.5 months (95% confidence interval 6.3-19 months). Thirty-eight percent of the patients were alive at one year and 19% at two years. CONCLUSION: The combination of 5-fluorouracil, leucovorin and alpha interferon as administered in this phase II study did not result in enhanced response rate or survival. However this regimen was associated with considerable toxicity. PMID- 10803324 TI - Review: the application of dendritic cell-derived exosomes in tumour immunotherapy. AB - Cancer arises from the aberrant proliferation of a single transformed cell. This population acquires the ability to metastasis. An effective way to remove cancer cells from the body is to activate tumour-specific cytotoxic T cells (CTL). Activation of naive T cells depends on the unique antigen presenting capacity of DC. Activated tumour antigen-specific CTL can destroy cancer cells without harm to normal tissue. Their ability to stimulate antigen specific T cell responses makes DC attractive candidates to potentiate anti-tumour immunity. Several studies have demonstrated the efficacy of DC based anti-tumour immunotherapy and the goal now is to optimise immune responses induced by DC, so that effective strategies in treating cancer may be realised. One way to do this is to identify DC characteristics which make them more effective in T cell stimulation. Another is to use exosomes, the antigen presenting vesicles secreted by DC, in order to induce potent anti-tumour immune responses. The non-cellular nature of exosomes offers several advantages for use in tumour immunotherapy. PMID- 10803325 TI - Effect of red blood cells on the antitumor activity of oxycellulose. AB - The antitumor activity of oxycellulose to MH134 tumor cells, evaluated with microscopic viable cell count, was suppressed by murine red blood cells (MRBCs) in a dose-dependent manner. The antitumor activity of oxycellulose was suppressed by MRBCs also when evaluated with 3H-TdR uptake. However, that of mitomycin C or 5-fluorouracil was not suppressed by MRBCs. On the other hand, the antitumor activity of oxycellulose was suppressed by MRBCs also when evaluated with 51Cr release assay. At that time, the antitumor activity of cisplatin to MH134 tumor cells were not suppressed by MRBCs. The antitumor activity of oxycellulose to MH134 tumor cells was suppressed by the membrane or the cytosol fraction of the lysate of MRBCs as well as intact MRBCs. When heated at 56 degrees C for 30 min, MRBCs were unable to suppress the activity of oxycellulose. It was examined whether or not proteins related to MRBCs (hemoglobin, catalase, superoxide dismutase or taurine) suppressed the activity of oxycellulose. They did not suppress the activity of oxycellulose. PMID- 10803326 TI - Apparent pokeweed mitogen cure of metastatic gum melanoma in an older dog. AB - Immunotherapy with plant mitogens has been of increasing interest to both authors, although their mutual attraction to these substances over the past two decades occurred independently and has taken divergent pathways from the start. Because of their clinical unavailability, Wimer's efforts have been confined to writing theoretical concepts on potential applications of the mitogens focusing on the L4 isolectin of PHA. Alternatively, Mann has worked actively with laboratory and experimental research involving PWM that he has extracted himself. As a sequel to a pilot study of PWM cancer treatment in pets whereby he supplies the mitogen, protocol, and data sheets and veterinarians supervise the administration, Mann acquiesced to widespread requests for inclusion in an extended investigation. Unfortunately, this arrangement has left Mann with little control over data submission, and until recently, only verbal reports have been received. The recent documentation of a three-year remission and apparent cure of gum melanoma metastatic to regional and hilar lymph nodes and to the lungs in an aged dog following PWM therapy has prompted this case report. The incredible response has inspired more aggressive attempts to obtain information from other recipients of the mitogen. The small total dose of 300 micrograms inducing a remission does speak for the remarkable potency of PWM that may be 500 or more times that of PHA. However, the possibility that melanoma is a uniquely responsive tumor is suggested by a 1971 article indicating complete remission of metastatic melanoma in an 18-year-old lad to an unexpectedly low total dose of 50 mg PHA. Preliminary reports on other tumors treated with PWM indicate some impressive responses have occurred, although they also suggest that Mann's regimen may sometimes require adjustments in dosages and other variables previously reviewed in theoretical PHA-L4 models, to which this case report now contributes validity. PMID- 10803327 TI - Mitogen hypotheses revisited: enhancement of allograft tolerance and replacement of transplantation procedures in treating cancer. PMID- 10803328 TI - Clinical experience with recombinant human thyrotrophin (rhTSH) in the management of patients with differentiated thyroid cancer. AB - The purpose of this work was to gain clinical experience with and to identify the optimal conditions for the use of recombinant human TSH (rhTSH, commercially available as Thyrogen) in the management of patients with differentiated thyroid cancer (DTC). The study involved 22 patients for a total of 27 administration cycles of rhTSH, for either diagnostic (in 19 instances) and/or therapeutic purposes (in 8 instances). There were 19 patients with papillary cancer (follicular variant in 4, columnar variant in 1) and 3 patients with follicular cancer (1 Hurtle cell variant). All patients had previously undergone total thyroidectomy and 1-5 cycles of 131I-therapy. Thyrogen was administered i.m. according to the suggested protocol: 0.9 mg i.m. on days 1 and 2, radioiodine on day 3. Peak serum TSH levels between 68-237 microIU/mL were observed after rhTSH administration; these were on average 65% higher, on a patient-by-patient basis, than peak serum TSH observed after conventional withdrawal of thyroxine treatment in 19 patients, while in 3 patients they were 28% lower, but still in the potent stimulation range (86-94 microIU/mL). There was general agreement between imaging results obtained under rhTSH stimulation and those obtained on prior occasions during thyroxine withdrawal, although radioiodine uptake was interpreted as less intense following Thyrogen administration. Of 18 patients undergoing rhTSH administration for diagnostic purposes, 11 patients had a negative radioiodine whole-body scan (WBS) and 7 had a positive WBS. Three of the WBS-negative patients were shown to be actually affected by tumor recurrence, respectively by PET with [18F]FDG (in 2 cases) and by post-131I therapy scan. Serum thyroglobulin (hTg) increased to abnormal levels following rhTSH stimulation in 3/7 of the WBS positive patients as well as in 1/11 WBS-negative patients. In 3/7 WBS-positive as well as in 3/11 WBS-negative patients, serum hTg progressively rose under rhTSH stimulation, yet still remaining below 3 ng/mL. Post-131I therapy scans following Thyrogen administration showed good radioiodine uptake in 7/8 patients, the single unsuccessful case being most likely due to expansion of the iodine pool because of recent use of an iodinated contrast medium. The overall results show the feasibility and practical advantages of employing rhTSH stimulation in the general clinical setting rather than thyroxine withdrawal in the management of DTC patients. Caution should be raised on the interpretation of the serum hTg response to such potent but short-lived TSH stimulation. PMID- 10803329 TI - History of the operative management of pectus deformities. AB - This article examines the history of the operative management of pectus deformities, tracing the surgical innovations of such pioneers as Drs. Ochsner, Ravitch, Sanger, and Wada. Guidelines for the selection of patients for operative correction are also discussed. PMID- 10803330 TI - Embryology of the sternum. AB - Development of the sternum during the growth of the embryo is described. Anterior body wall defects in the thoracic region may be severe, leading to ectopia cordis, or mild, as in skin-covered sternal clefts. The embryologic basis for other sternal abnormalities, such as pectus excavatum and pectus carniatum, is not clear; however, abnormalities of rib morphogenesis and growth are the most likely causes. PMID- 10803331 TI - Cardiopulmonary effects of anterior chest wall deformities. AB - Many studies have been conducted that evaluate the cardiopulmonary effects of pectus excavatum because of the general clinical impression that post-repair patients have better exercise tolerance and vigor. The results of these studies have been variable during the last three decades, and although restrictive lung volumes are often present in subjects with pectus excavatum, they are not invariably improved by repair. Enhanced cardiac performance may be partially responsible for improved function. This article reviews the results of these studies. PMID- 10803332 TI - Repair of pectus excavatum. Anesthetic considerations. AB - The majority of patients undergoing pectus excavatum repair can be safely anesthetized with routine clinical preoperative evaluation and chest radiography. Electrocardiography, echocardiography, arterial blood gas, and pulmonary function tests should be reserved for patients with suspected coexisting conditions, such as mitral valve prolapse, skeletal abnormalities, Marfan's syndrome, and pulmonary disease. The choice of surgical repair and the advisability of placement of an epidural catheter or intercostal blocks for perioperative pain management should be discussed with the surgical team. Postoperatively, patients should be carefully monitored for signs of tension pneumothorax and pneumonia. PMID- 10803333 TI - Cleft sternum and sternal foramen. AB - Cleft sternum is a rare congenital defect of the anterior chest wall and is the result of a failed midline fusion of the sternum. Depending on the degree of separation, there are complete and incomplete forms. Its clinical significance is that it leaves the heart and great vessels unprotected. Association with craniofacial hemangiomas and omphalocele is common. Prenatal diagnosis by ultrasonography is possible. Surgical correction should be performed during the neonatal period when the direct suturing of the sternal halves is possible and the thorax can accommodate the thoracic viscera. At an older age, surgical repair is feasible, but it may require additional measures, such as sliding chondrotomies of the adjacent costal cartilages and notching of the sternal bars, to facilitate the approximation. Lung herniation at the base of the neck should be corrected by uniting the cervical muscles. The first postoperative day is the most critical because of acute reduction of the mediastinal space. Long-term results are satisfactory. Sternal foramen is a congenital oval defect at the lower third of the sternum that is asymptomatic and could be detected by CT scanning. The awareness of the anomaly is important in acupuncture practice because of the danger of heart damage. PMID- 10803334 TI - Surgical treatment of pectus excavatum. AB - In conclusion, the following points are reemphasized: 1) The abnormal (either depressed or protruding) cartilages should always be resected. This resection, especially in pectus excavatum abnormalities, should not be overdone because the highest point that the chest wall and sternum can be elevated to with these types of operations is only the level of the most anterior rib and the thickness of the sternum itself. 2) Marlex mesh is an ideal material to support the sternum in its corrected position. It is strong and holds well until the chest wall solidifies. Also, it is resistant to infection and it may be left in place permanently. The application of different metallic splints, rods, and so on, as well as costal allografts, was found to be absolutely unnecessary. 3) Intercostal strips detached from the sternum may be left in place. Also, the surgeon should not waste time in performing a meticulous "classic" subperichondrial resection of the cartilages and ribs but should just leave enough perichondrium and periosteum behind to ensure the regeneration of the ribs. For the same reason, a segment of the most lateral portion of the cartilage should be left in continuity with the ribs. 4) It is strongly recommended that in excavatum anomalies, one of the pleural cavities should be deliberately opened and wide communication established between the pleural and the retrosternal space; the entire operative area should be drained for a day or two using an intracostal water-sealed catheter. This will make the use of any other subcutaneous or mediastinal drainage devices unnecessary and will ensure appropriate drainage of blood or serum. It is also recommended that the resection of the cartilages should be done on the left side first, where inadvertent entering of the pleura is less likely because of the backing of the pericardium. If it happens, drainage of the right hemithorax is not necessary. Carinatum anomalies are handled with subcutaneous drainage. 5) To confirm appropriate results, the chest should be carefully inspected after closure of the skin, and flaws, if they exist, should be corrected right then. Also, surgical repair of all pectus anomalies, especially excavatum deformities, should be supplemented in due time with an appropriate exercise program. Swimming and weight lifting are especially useful. 6) We found that the age limit imposed on small children by some authors is unnecessary, and as a matter of fact it is preferable to operate on children at an early age, around 2 years, because of commonly existing psychologic problems at a later age. The author advises restraint in operating on individuals past the teenage years unless the deformity is physiologically restricting. 7) The jury is still out regarding procedures using limited exposure and that do not use transverse sternotomy to correct the depressed or elevated sternal axis. 8) The usage of cosmetic procedures, or in other words, operations that do not correct the anomaly of the bony chest wall but use various implants as camouflage, should be restricted to cases of moderate excavatum anomalies in late teenage patients and to adults without cardiorespiratory symptoms. PMID- 10803335 TI - Ectopia cordis. AB - Ectopia cordis is a rare congenital defect in which the heart is placed externally on the surface of the chest. This article describes the embrylogic events that lead to the various classifications of the defect and how they possibly explain the process of its unusual occurrence in children. While survival in some cases is possible, the ultimate repair is difficult and survival is rare. The most extreme forms of ectopia cordis, especially those with intracardiac defects, have a poor prognosis. Several cases of this rare anomaly with primary repair and staged repair are discussed. PMID- 10803336 TI - Turnover procedure. AB - Sternal turnover can be considered an orthotopic bone transplantation in reverse position. It yields immediate stability of the chest and good cosmetic results. Procedures using "pedicled plastrons" (rectal muscles, internal mammary arteries) have been reported; a large series of 300 cases, however, shows satisfying results with free grafts. Technetium-99m isotope studies yielded visualization of the plastron 1 year after sternal turnover. After redo-operations, histopathologic studies gave nearly normal sternal bone marrow findings. PMID- 10803337 TI - Minimally invasive pectus surgery. AB - The technique of minimally invasive repair of pectus excavatum is a new operation that allows for repair of this deformity without any cartilage resection or sternal osteotomy. The procedure has revolutionized the management of pectus excavatum. The innovative incorporation of thoracoscopic techniques and small but important modifications to the technique have made this operation very effective and safe. PMID- 10803338 TI - Reconstruction of congenital chest wall deformities using solid silicone onlay prostheses. AB - Congenital chest wall deformities include five types: pectus excavatum (funnel chest), pectus carinatum (pigeon breast), Poland's syndrome, defects of sternal fusion, and miscellaneous dysplasias and skeletal disorders. Of these five types, two, pectus excavatum and Poland's syndrome, are defects of the skeletal chest wall. These two specific anomalies comprise the vast majority of congenital defects of the chest wall and, as depression deformities, are readily amenable to surgical correction. PMID- 10803339 TI - Surgical treatment of pectus carinatum. AB - Pectus carinatum, just like its sister deformity pectus excavatum, is a condition with an undefined developmental mechanism and debated surgical techniques. Elongation of the costal cartilages and elongation and anterior displacement of the sternum characterize the different varieties of pectus carinatum. Repair of the anomaly involves positional correction as well as the shortening of the sternum and the maintenance of its corrected position by action of the rectus abdominis and pectoralis muscles. PMID- 10803340 TI - Pouter pigeon breast. AB - Pouter pigeon breast is a rare congenital deformity of the chest characterized by a protrusion of the manubriosternal junction and premature sternal ossification. The adjacent costal cartilages, usually from the second to fifth, also protrude. One third of the patients with pouter pigeon breast presented with concomitant depression of the lower third of the sternum. Several cardiovascular abnormalities have been associated with premature sternal ossification, with ventricular septal defect being the most common. All patients with pouter pigeon breast should undergo echocardiography in search of occult cardiac lesions. The measurement of the angle of Louis on lateral chest radiograms allows the objective assessment of the deformity and appreciation of the sternal fusion. Surgical correction includes the wide wedge transverse sternotomy at the angle of Louis and subperichondrial resection of the adjacent costal cartilages. When the deformity is part of a syndrome or in cases of severe depression of the lower sternum, we recommend the technique using Marlex mesh support as described by Robicsek. Preferable age for repair is 5 to 7 years old. Long-term outcomes are encouraging. PMID- 10803341 TI - Poland's syndrome. AB - Poland's syndrome is characterized by hypoplasia or absence of the breast or nipple, hypoplasia of subcutaneous tissue, absence of the costosternal portion of the pectoralis major muscle, absence of the pectoralis minor muscle, and absence of costal cartilages or ribs 2, 3, and 4 or 3, 4, and 5. Clinical manifestations of Poland's syndrome are extremely variable and rarely are all the features recognized in one individual. Depending on the physician's specialty and the referral pattern, a variable incidence of the anomalous defects is recognized. Syndactyly or bony abnormalities of the forearm are seldom treated by thoracic surgeons. PMID- 10803342 TI - Simultaneous pectus and open heart surgery. AB - Various surgical approaches to pectus excavatum repair concomitant with surgery have been recommended. In this article the authors describe their approach to the problem that they applied in 1989 and onward, successfully, in six consecutive patients. The favorable early and long-term results of these cases illustrate that the simultaneous correction of pectus excavatum and the underlying diseases of the ascending aorta, aortic arch, and the heart can be performed successfully even in emergency situations. The technique recommended provides good cosmetic results and a stable chest wall. It is well applicable in patients of adult age. PMID- 10803343 TI - Complications of surgery for pectus excavatum. AB - The article describes the various acute and late complications of surgery for pectus excavatum. Because the acute complications are well known and easily managed and the late complications can be prevented, operative correction of pectus excavatum can be recommended to parents and their children with severe deformities with very little risk and a realistic expectation of good long-term correction. PMID- 10803344 TI - Scoliosis in children with anterior chest wall deformities. AB - An association between anterior chest wall deformities and scoliosis is described in the literature but is poorly defined. It appears that only approximately 4% to 5% of patients with severe anterior chest wall deformities have scoliosis of sufficient magnitude to warrant evaluation and observation by a spinal deformity physician. The relationship between anterior chest wall deformity and scoliosis is most clear in patients with Marfan syndrome. Marfan patients with scoliosis are at high risk for progression of deformity to unacceptable levels and have historically not responded well to brace therapy. The causes of anterior chest wall deformity and scoliosis remain unclear, although unbalanced cartilage growth has been proposed as a potential cause of both. Some have proposed that chest wall deformity may result in asymmetric spinal forces and subsequent scoliosis secondary to altered spinal growth. Because of the association between pectus deformities and scoliosis, patients with anterior chest wall deformities should be carefully examined for signs of scoliosis and have screening radiographs if indicated. Young patients in particular should be referred to a spinal deformity physician, because it is clear that patients who present with spinal deformity prior to age 5 years are the ones at risk for adverse cardiopulmonary sequelae related to spinal deformity. Most children with scoliosis and anterior chest wall deformities need only observation of the scoliosis. In more severe cases, the management of spinal deformity traditionally involves the use of bracing to attempt to control curves of lesser magnitude and surgical management for unacceptable deformities or curves of magnitudes too great for brace treatment. There are no studies that specifically address the use of these methods in patients with both anterior chest wall deformities and scoliosis. The management of scoliosis in patients with anterior chest wall deformities follows treatment principles outlined for idiopathic scoliosis patients. PMID- 10803345 TI - What to expect from MRI in the investigation of the central nervous system? AB - Functional magnetic resonance imaging (fMRI) has appeared as a new tool that is very powerful for cognitive neuroscience, offering the potential to look at the dynamics of cerebral processes underlying cognition, non-invasively and on an individual basis. Work remains to be done to optimize the technique and to better understand its basic mechanisms, but one may expect to build in a foreseeable future a functional list of the main brain cortical networks implicated in sensory-motor or cognitive processes. Still, the real understanding of brain function requires direct access to the functional unit consisting of the neuron, so that one may look at the transient temporal relationships that exist between largely distributed groups of hundreds or thousands of neurons. Furthermore, communication pathways between networks, which are carried by brain white matter, must be identified to establish connectivity maps at the individual scale, taking into account individual variability resulting from genetic factors and cerebral plasticity. In this respect, MRI of molecular diffusion is very sensitive to water molecular motion and, thus, to tissue dynamic microstructure, such as cell size and geometry. Preliminary data suggest that diffusion MRI visualizes dynamic tissue changes associated with large neuronal activation and space orientation of large bundles of myelinated axons in the white matter. PMID- 10803346 TI - [Persistence and resurgence of sleeping sickness caused by Trypanosoma brucei gambiense in historic foci. Biomathematical approach of an epidemiologic enigma]. AB - Since the end of the 19th century, historic endemic foci of Trypanosoma brucei gambiense sleeping sickness have proven very persistent. A five-compartment mathematical model with open vector populations was developed in order to study the dynamics of this disease in Central Africa. Of particular interest is the rate at which the disease spreads or goes to extinction at the beginning of an epidemic outbreak. A measure of this rate is the initial halving/doubling time T(o) of the numbers infected; T(o) is a doubling time when the basic reproduction number Ro > 1 and a halving time when Ro < 1. For realistic parameter values, T(o) can be quite large (i.e. several years or even decades) which corresponds to a persistent low-level endemic brought about by an Ro either just above 1 (slow spread) or just below 1 (slow extinction). A resurgence of historical foci can then be caused by a small shift in parameter values that brings Ro well above 1 and decreases T(o). In addition, when Ro is less than 1 (in the absence of vector migrations), simulations show that a very small percentage of infected immigrant flies can bring about high prevalence rates in the human population. The model is validated with field data from historical Congolese, Central and West African foci of the past. PMID- 10803347 TI - Seasonal variations of thyroid hormone levels in wild fish. AB - Variations in plasma thyroxine (T4) and triiodothyronine (T3) contents of pike, pikeperch, perch, bream and chub from a large reservoir (Pareloup, South of France) were investigated before and during the progressive emptying of the reservoir. Fish were caught by gill nets, except at the last time of emptying when controls were collected with landing nets. Both iodothyronine levels showed desynchronized seasonal cycles resulting in higher T3/T4 ratios in fish stressed by gill-netting in early summer. The aforementioned results principally emphasized the prevailing influence of the technique of catching on eliciting stress responses and subsequent thyroidal impairment in all five species. However, the much higher amounts of T4 and/or T3 observed in specimens caught in the later stages of draining gave evidence to suggest that the progressive alteration in habitat might also have indirectly affected the thyroidal status of these fish, this latter result leading to the consideration of fish plasma thyroid hormones as possible endocrine biomarkers of chronic perturbation. PMID- 10803348 TI - [First report of Aedes albopictus (Skuse, 1984) in metropolitan France]. AB - The first record of Aedes albopictus in metropolitan France has been made in a village of Orne (Basse-Normandie). A few larvae were collected in October 1999, in the used tire stock of an important tire recycling company, importing in particular from the USA and Japan. Reproduction of the species has taken place in France, and the environmental conditions make the implantation of the species probable. PMID- 10803349 TI - Seabirds as monitors of upper-ocean thermal structure. King penguins at the Antarctic polar front, east of Kerguelen sector. AB - The main objective of this work was to assess the potential of diving birds to monitor the hydrographic features near the Antarctic polar front. We compared the temperature/depth profiles recorded by instrumented King penguins Aptenodytes patagonicus at Kerguelen Islands (South Indian Ocean) with the oceanographic and remote sensing (satellite) data available for the same area during the same season. The birds were equipped with time/depth/temperature recorders or Argos transmitters. In addition, two birds were instrumented (of which one successfully) both with a time/depth/temperature recorder and an Argos transmitter. King penguins foraged as far as 400 km from the coast, in water masses with a vertical temperature structure characteristic of the region just south of the polar front. The temperature/depth profiles recorded throughout the dives (up to 270 m) revealed a pronounced thermocline. A three-dimensional distribution of water temperature was reconstructed. Comparison with previous hydrographic data shows a high correlation. Instrumented predators may therefore usefully and cheaply complement the database provided by conventional hydrographic surveys and remote sensing, especially in distant and rough areas such as the Southern Ocean. PMID- 10803350 TI - The attendance cycles of the Cory's Shearwater Calonectris diomedea borealis on Selvagem Grande. AB - In the Cory's Shearwater Calonectris diomedea borealis of Selvagem Grande (30 degrees 09'N, 15 degrees 52'W), the visits the birds pay to their colonies are cyclic throughout the breeding period. The number of visitors varies regularly between a minimum and a maximum following a sinusoid, the half-period of which is 4.8 +/- 1.5 d (2-8 d). No variation exists according to the status of the birds (breeders or non-breeders) or to the stage of the breeding cycle (pre-egg, incubation or chick rearing) and moon phases have no influence. Among Procellariids, such attendance cycles are related to the length of the pre-egg stage. They are absent when it is short (less than 50 d), present when it is long (more than 60 d), and indifferently present or absent when it is intermediate (50 60 d). Considering the comparative scarcity of the visits of the birds to the colonies during the pre-egg stage, these cycles are likely to play a part essentially in the synchronisation of individuals in colonies and of mates in pairs. PMID- 10803351 TI - Role of vegetable tannins in habitat selection among mosquito communities from the Alpine hydrosystems. AB - The role of vegetable tannins in habitat selection among mosquito communities in Alpine hydrosystems was investigated through ecotoxicological comparison of 19 arthropod species characteristic of 12 breeding sites known for their abiotic environmental factors and their different riparian vegetation. The toxicity of tannins was experimentally compared among species representative of both the dipteran fauna and the crustacean fauna associated with the mosquito breeding sites. Bioassays using tannic acid solutions at concentrations from 0.1 to 11 mM separated the dipteran taxa into five groups of differential sensitivity and the crustacean taxa into four groups. The different levels of sensitivity among taxa were correlated with the various amounts of total phenolics and tannins found in the most prominent plant types associated with the different breeding sites. This suggested that tannins and, more generally, phenolic compounds may be involved in plant-arthropod interactions in Alpine hydrosystems. PMID- 10803352 TI - [Comparison of genetic diversity (RAPD) of ex situ collections and natural populations of Naufraga balearica Constance & Cannon]. AB - Naufraga balearica Constance & Cannon (Hydrocotyloideae) cultivated in the Botanical Gardens of Lyon, Brest and Porquerolles stem from two or three shoots collected in Corsica in 1981. The genetic diversity of these plants was evaluated using RAPD markers (random amplified polymorphic DNA). It was compared with the diversity found in individuals collected from five natural sites in Majorca. Only a few patterns were present in the collections derived from the Corsican shoots. The plants kept in the Botanical Gardens appeared to be of clonal origin: most individuals (81%) showed a 'dominant pattern'. In contrast, nearly all individuals sampled in the natural populations of the Balearic Islands exhibited a unique pattern. The five populations appeared genetically distinct; the individuals probably resulted from cross-fertilizations. The cultivated Corsican plants from Lyon, Brest and Porquerolles appeared genetically closely related to the individuals sampled in the population of Cala San Vicente in Majorca. The spontaneity of this paleoendemic in Corsica was discussed. PMID- 10803353 TI - Distribution and dynamics of Posidonia oceanica beds along the Alberes coastline. AB - The location and dynamics of Posidonia oceanica beds were studied in the marine natural reserve of Cerbere-Banyuls-sur-Mer (eastern Pyrenees, France), through the mapping of bottom assemblages (transects and image processing) and the monitoring of fixed markers installed at the Posidonia oceanica lower limit. The surface area occupied by continuous beds is relatively limited and localized, whereas the presence of important areas of dead 'matter' would seem to indicate that bottom cover by Posidonia oceanica was more extensive in the past. Although the limited extension of the seagrass beds seems to be linked to both a high level of turbidity and severe hydrodynamic conditions, it is hypothesized that the regressions observed are due to lack of sediment. PMID- 10803354 TI - [Thermal anomaly in waters of the Gulf of Marseilles during the summer of 1999. A partial explanation of the mortality of certain fixed invertebrates?]. AB - During summer 1999, a massive mortality of attached benthic invertebrates was reported at several points along the French Mediterranean coast, east of the Rhone River. Such a wide geographical extension suggested the occurrence of a climatic anomaly as a direct or an indirect cause of this phenomenon. The systematic observations of surface water temperature (1 record/30 min) and vertical thermal structure from 0 to 55 m (semi-monthly frequency) performed by the service of observation of centre d'oceanologie de Marseille since 1994 in the Gulf of Marseilles (northwestern Mediterranean) compared with historical data obtained for the summer periods (June till October) indicates a thermal anomaly during summer 1999. Although the surface temperatures do not strictly show exceptional values, the stability during long periods of high temperatures (24.1 degrees C +/- 0.5 in September, n = 1,018) and the thermal structure of the water column are remarkable. The vertical temperature profiles of 1999 reflect a general warming of 2-3 degrees C in the water layer overlying the thermocline, accompanied by its deepening (up to 30-40 m). This situation can be explained by the scarcity of NW winds with a marked drop of their relative frequency during July and October 1999 (13-27% versus 42%, mean value calculated from the previous 12 years) and the short duration of these windy periods (between 30 min and 5 h as a mean). Parallel to this, a large number of no wind registrations (125-250 h/month as a mean) were observed during this period. A similar thermal stratification, of lesser amplitude, associated with comparable meteorological conditions was also observed in 1997. The hypothesis that this summer 1999 temperature anomaly could constitute a partial explanation for the recorded massive mortality is discussed. PMID- 10803355 TI - Developing a new model for non-insulin dependent diabetes mellitus (NIDDM) by using the Philippine wild mouse, Mus musculus castaneus. AB - The Philippine wild-caught castaneus mouse (Mus musculus castaneus) and laboratory mouse (C57BL/6J: B6) were used to develop a new non-insulin dependent diabetes mellitus (NIDDM) model. Offspring from the cross between a wild male and B6 female were backcrossed to the sire. One male which exhibited highest fasting hyperglycemia (190 mg/dl) among eighty-seven backcross offspring was selected at 10 weeks of age, and crossed with a B6 female to comprise the fundamental stock (F0). Thereafter, full-sib mating was performed to develop a new inbred strain named CBD (Castaneus-B6 diabetic) mouse. Mice with relatively higher fasting hyperglycemia among F0 and F1 generations were selected for breeding. From the F2 generation, mice were defined as diabetic when blood glucose levels exceeded 200 mg/dl at 120 min in intraperitoneal glucose tolerance test (IPGTT) at 10 weeks of age, and have been selectively bred. The incidence of diabetic males from the F3 F6 generation fluctuated 45-75% at 10 weeks of age and 59-72% at 20 weeks of age. Diabetic males had about two-fold higher fasting glucose and insulin levels than B6 males. Glucose-stimulated insulin secretion was impaired in diabetic CBD mice compared to B6 males at 20 weeks. Moreover, diabetic mice had slight obesity compared to B6 mice. These facts indicated that diabetic features of CBD mice resemble NIDDM in humans. The CBD strain, characterized by high incidence and early onset of diabetes with mild obesity would be of value as a new NIDDM model. The method, utilizing wild castaneus mouse of different origin from laboratory mice, maybe useful in the development of other animal models. PMID- 10803356 TI - Histological analysis of murine colitis induced by dextran sulfate sodium of different molecular weights. AB - In this study, we examined the relationship between the molecular weight of dextran sulfate sodium (DSS) and the features of colitis in a DSS-induced mouse model of human ulcerative colitis. DSS at three different molecular weights, 5 kD, 40 kD and 500 kD, was used in this study. DSS was administered in drinking water at 5% (w/v) to 6-7-week-old female BALB/c mice. After 7 days of treatment with DSS, the large intestine was examined histopathologically. Colitis was characterized by a loss of crypts, infiltration of inflammatory cells into the mucosa and submucosa, edema of the submucosa, erosion and ulceration and was observed in mice given the 5 kD and 40 kD forms but not the 500 kD. In the 5 kD group, colitis was observed predominantly in the cecum and upper colon. Colitis in the 40 kD group was more severe than that in the 5 kD group, and in the 40 kD group it was more severe in the lower colon than in the upper colon. These findings suggest the molecular weight of DSS to be an important factor in the murine model of colitis. PMID- 10803357 TI - Transplantation of cryopreserved mouse, Chinese hamster, rabbit, Japanese monkey and rat ovaries into rat recipients. AB - Partial ovaries from mice, hamsters, rabbits, Japanese monkeys and rats have survived deep-freezing and returned to a normal morphological state after being thawed and transplanted into the rat uterine cavity. This report describes the ice-free cryopreservation of mouse and other ovaries at -196 degrees C by vitrification. The vitrification solution was based on the solutions reported by Rall & Fahy [16]. After ovaries had been exposed to the vitrification solution, they were frozen, with their suspending medium, by liquid nitrogen. After freezing, the ovaries were thawed in 37 degrees C water. The viability of the previously frozen ovarian tissue was tested by transplanting it into the uterine cavity of pseudopregnant rats. Seven days after transplantation, the ovaries were removed with the rat uterus, and stained with haematoxylin and eosin for histological examination. Survival of the frozen-thawed the ovaries in the rat uterine cavity demonstrates that these ovaries can tolerate exposure to osmotic dehydration and vitrification in a concentrated solution of cryoprotectant and are then immunologically acceptable to the uterine cavity. PMID- 10803358 TI - Electrophysiological properties of ventricular muscle obtained from spontaneously diabetic mice. AB - The electrophysiological properties of cardiac muscle in KK/Ta mouse (hereafter referred to as KK mouse), an animal model of human non-insulin-dependent diabetes mellitus, were investigated, and the findings compared with those obtained from a non-diabetic control mouse (C57BL/6J mouse; referred to as B6 mouse). The ages of the B6 mice were 23.9 +/- 5.4 weeks (n = 24) and those of the KK mice used were 25.7 +/- 10.8 weeks (n = 34). The KK mice had mild obesity, hyperglycemia and hyperinsulinemia. Ventricular muscles from both mice were examined by light microscopy. Partial myocardial fibrosis and filament disorder in the ventricular muscles were found only in the KK mice. The resting membrane potential of the ventricular muscle was less negative in the KK mice than in the control mice. The maximum rate of rise in the upstroke of the action potential was significantly decreased in the KK mice compared with that of the control mice. These suggest a decrease in a time-independent K+ current (IK1) in the KK mice. The duration of the action potential (APD) at all levels of repolarization was significantly longer in the KK mice than in the B6 mice. A blocker of transient outward current (I(to)), 4-aminopyridine, significantly prolonged the APD of the B6 mice, but failed to prolong it in the KK mice, suggesting that Ito in the diabetic mice is very small. A Ca2+ channel blocker, CoCl2, dramatically lengthened all levels of APD in both groups, suggesting that there is no difference between B6 mice and KK mice in L-type Ca2+ current via Ca2+ channels. These suggest the malfunction or deficiency of ionic channels which carry, at least Ito and IK1 in diabetic mice. PMID- 10803359 TI - Effects of Eurycoma longifolia Jack (Tongkat Ali) on the initiation of sexual performance of inexperienced castrated male rats. AB - We studied the effects of Eurycoma longifolia Jack, commonly known as Tongkat Ali in Malaysia, on the initiation of sexual performance and the weights of sexual accessories in inexperienced castrated male rats. The doses of 200, 400 and 800 mg/kg body weight, which were extracted from E. longifolia Jack, were orally administered to the rats twice daily for 10 days prior to the tests and continued throughout the test period. Testosterone was used as a positive control after injecting 15 mg/kg daily subcutaneously for 32 days. Results showed that E. longifolia Jack produced a dose-dependent increase in sexual performance of the treated animals, but the E. longifolia Jack groups showed lower sexual performance in mounting, intromission and ejaculation than the testosterone group. Further results also showed that E. longifolia Jack promoted the growth of both ventral prostate and seminal vesicles as compared with the control, but the growth of sexual accessories at 800 mg/kg of butanol, methanol, water and chloroform fractions of E. longifolia Jack was less than that of testosterone treated group. The present study therefore gives further evidence of the folkuse of E. longifolia as an aphrodisiac. PMID- 10803360 TI - Decontamination of human xenotransplantable tumor with mouse hepatitis virus by implantation in nude rat: a case report. AB - A human tumor xenograft contaminated with mouse hepatitis virus (MHV) was implanted in a nude rat in order to decontaminate the tumor line. The decontamination failed in the first trial, but succeeded in the second trial. The difference between the two trials was the duration of implantation of the tumor in the nude rat, i.e., 12 days in the first and 24 days in the second trial. Duration of implantation might be a factor in the decontamination of transplantable tumors infected with MHV by passaging in the nude rat. PMID- 10803361 TI - Mapping of the beige (bg) gene on rat chromosome 17. AB - The rat beige (bg) autosomal recessive gene, causing Chediak-Higashi Syndrome (CHS) in rat, was mapped on Chr 17 by using synteny of rat to mouse and humans. The linkage between the beige gene and PCR-amplified microsatellite markers in (DA-bg x BN)F1 x DA-bg backcross progeny was analysed. The recombination frequency was 9.5% between Prl and Acrm and 19.1% between Acrm and bg. The proposed order of three genes is Prl-Acrm-bg. This rat bg gene was confirmed to be homologus to the beige (bg) gene of mouse located on Chr 13 and the CHS (Lyst) gene of man located on Chr 1 (1q43). PMID- 10803362 TI - Blunt cell growth potential in carcinogen resistant inbred DRH rats. AB - A carcinogen-resistant inbred strain DRH/Sea has been developed from the Crj:Donryu strain. The rats had a very low incidence of liver tumors when they were fed diets containing a hepatocarcinogen such as 3'-methyl-4-dimethylamino azobenzene (3'-Me-DAB). Despite using 3'-Me-DAB during the stage of selection, the DRH/Sea rats developed normally, reproduced and did not have any spontaneous tumor in the lung, liver or uterus at over 1 year of age. Although their growth curves were similar to the Crj:Donryu rats, the progression of polyploidization in the liver was significantly delayed when compared with Crj:Donryu rats. Mitogenic changes that occurred in the liver caused by either 3'-Me-DAB or lead nitrate were less significant in the DRH rats than in Crj:Donryu rats. Furthermore, the growth rate of cultured fibroblasts derived from the DRH rats was slower than that of Crj:Donryu rats. These results, together with our previous results, suggest that slow growth potential is present under certain conditions in DRH rats. These findings may explain partly the meaning of the different susceptibility to hepatocarcinogens. PMID- 10803363 TI - Characterization of a novel polycystic kidney rat model with accompanying polycystic liver. AB - A polycystic kidney rat model is being established from a Crj:CD (SD) rat strain. Unlike existing animal models of polycystic kidney disease, this mutant rat has a completely polycystic liver. Mating experiments revealed that the phenotype is controlled by an autosomal recessive gene. We propose that this gene be tentatively called the "rpc" gene. PMID- 10803364 TI - Influence of castration on development of the thymus in neonatal male rats. AB - The influence of castration on the development of the thymus in neonatal rats was studied to elucidate when after birth the thymus comes under inhibitory regulation by the testis in rats. The relative and absolute weights of the thymus were measured five days after castration these cases. No excessive changes in the weights of the thymus with castration were observed by 31 days after birth. Significant changes in the thymus appeared in the relative weight at 36-day castration. The absolute weight of the thymus was also significantly increased after 41-day-castration. These findings suggest that in rats the inhibitory regulation of the thymus by testis development does not appear before at least 31 days of age. PMID- 10803365 TI - Sequence analysis of major structural proteins of newly isolated mouse hepatitis virus. AB - We have isolated the virus from a fecal pellet in the colon of a BALB/c mouse with X-linked immunodeficiency (xid) housed in a room in which there has recently been an epidemic due to mouse hepatitis virus (MHV) and designated it as the MHV TY strain. Sequence analysis of the MHV-TY strain was performed on major structural, spike (S), membrane (M) and nucleocapsid (N), proteins directly from PCR products. The comparison of nucleotide sequences of MHV-TY with other strains investigated so far revealed that all three structural proteins of the TY strain had some unique amino acid sequences among MHV strains which can be used as markers of this strain. PMID- 10803366 TI - The down-regulated in adenoma (Dra) gene encoding intestine-specific anion transporter maps to mouse chromosome 12. PMID- 10803367 TI - Protein structure analysis of today: proteomics in functional genomics. PMID- 10803368 TI - Proteome mapping by two-dimensional polyacrylamide gel electrophoresis in combination with mass spectrometric protein sequence analysis. AB - The high resolving power of two-dimensional polyacrylamide gel electrophoresis 2D PAGE and its full analytical and preparative potential have been described with special emphasis on reproducibility and standardization of protein spot patterns, enhanced protein detection sensitivity, and computer analysis database development. New methodologies for peptide mass fingerprinting, peptide, sequence, and fragmentation tagging have been highlighted. Major challenges associated with 2D-PAGE/mass spectrometric protein sequencing were outlined which need to be addressed in the future, including sample enrichment, use of alternative gel matrices, improvements in separation systems interfaced directly to the mass spectrometer, and design of high-sensitivity instruments with very high mass ranges. It is hoped that comparative studies to identify, quantitate, and characterize proteins differentially expressed in normal versus diseased cells would give insight into mechanisms of pathogenesis and allow the development of a way to control both the etiology and the course of diseases. PMID- 10803369 TI - Techniques for sample preparation including methods for concentrating peptide samples. AB - In the current era of proteomics two main analytical techniques are employed for protein identification. By far the fastest and most sensitive procedure for protein identification employs biological mass spectrometry, while de novo sequence analysis by classical Edman degradation is currently diminishing. In order to achieve the highest sensitivity for both techniques, great demands need to be put on sample preparation. In this paper we review three different aspects of protein sample preparation. Firstly, we discuss the use of polyacrylamide or agarose gel systems in which, during electrophoresis, proteins present in multiple primary gel pieces are eluted and simultaneously concentrated in a small secondary gel volume, whereby the overall sensitivity of Edman sequencing can be greatly increased. In a second chapter we review automation strategies occurring in the protein field which allow the automatic handling of multiple protein spots at the same time. In this context, we describe the use of auto-sampling techniques for further mass spectrometric studies and protein digestion robots allowing the simultaneous preparation of tens of gel-separated proteins. Finally we discuss various strategies for the preparation of biological peptide samples such as protein digests for both matrix-assisted laser desorption ionisation and electrospray ionisation mass spectrometry. PMID- 10803370 TI - Sample preparation by SDS/PAGE and in-gel digestion. AB - A convenient method for the isolation of samples for microsequencing or mass spectrometry utilizing polyacrylamide gel electrophoresis is described in detail, including pre-requisites for a successful outcome. Briefly, after visualization by e.g. Coomassie or silver, internal peptides are generated by in-gel proteolytic digestion, and after extraction, the corresponding protein is analyzed, i.e. the protein is identified by search in sequence data banks using a peptide sequence or a peptide mass map. Alternatively, a novel protein is detected, and an oligonucleotide probe for subsequent molecular biology work is synthesized, based on the obtained peptide sequence. One obvious advantage is that the procedure allows practically any protein to be studied. In-gel digestion has become the fundamental means of preparing samples in the many ongoing proteome projects. A few representative examples are given. PMID- 10803371 TI - High-resolution two-dimensional gel electrophoresis and protein identification using western blotting and ECL detection. AB - Two-dimensional gel electrophoresis has been the technique of choice for analyzing the protein composition of cell types, tissues and fluids and is a key technology in modern proteomics projects. Here we describe reproducible procedures for running isoelectric focusing and nonequilibrium pH gradient electrophoresis gels that are based on the carrier ampholyte technology originally described by O'Farrell. Moreover, we present a sensitive immunoblotting procedure that has been used routinely in our laboratory to determine the identity of hundreds of human proteins. PMID- 10803372 TI - Nanospray mass spectrometry in protein and peptide chemistry. AB - The introduction of electrospray in the mid-1980s revolutionised biological mass spectrometry, in particular in the field of protein and peptide sequence analysis. Electrospray is a concentration-dependent, rather than a mass-dependent process, and maximum sensitivity is achieved at low flow rates with high concentration, low-volume samples. This has lead to the development of nanoelectrospray, microelectrospray and related low flow-rate forms of electrospray which offer high sensitivity with low sample consumption. In this chapter the physical chemistry of low flow-rate electrospray is discussed, and a brief review of the types of low flow-rate electrospray interfaces is made. An indication of the performance obtainable on various instruments is given, along with some comments from the author's own experience of these techniques. PMID- 10803373 TI - MALDI-TOF mass spectrometry in protein chemistry. AB - Mass spectrometry has in the last decade been accepted as a key analytical technique in protein chemistry. It is now the preferred technique for identification of proteins separated by one- or two-dimensional polyacrylamide gel electrophoresis, i.e. in proteome analysis. It is the dominating technique for determination of posttranslational modifications in proteins. The two ionization techniques presently widely used in protein studies are matrix assisted laser desorption/ionization (MALDI) in combination with time-of-flight (TOF) mass analyzers and electrospray ionization (ESI) in combination with a variety of mass analyzers. In this chapter the principles and performance of MALDI-TOF mass spectrometry will be described as well as the application of this technique to a variety of applications. PMID- 10803374 TI - The chemistry of protein sequence analysis. AB - N-terminal sequence analysis by Edman chemistry continues to play an important role in the structural analysis of proteins and peptides. Improvements in the sensitivity of the method have been achieved mainly at the level of increasing the sensitivity of the on-line analysis of PTH amino acids by RP-HPLC (reverse phase high performance chromatography). Using microbore columns (0.8-1.0 mm), it is possible to run standards at the 0.5-1.0 pmol level and to sequence samples in the 1-5 pmol range. Due to constraints in current chromatographic methods, it is unlikely that further improvements in sensitivity will be achieved by this approach alone. Although alternative Edman reagents, including fluorescent chemistries, have promised to increase the sensitivity of sequencing into the low femtomole range, none of the methods have progressed into routine usage. These reagents and chemistries are critically evaluated in this review, and the problems which have prevented their further development discussed. Instrumental constraints are also considered. It is concluded that the development of more sensitive methods requires further research into both the chemistry and the instrumentation, and that alternative separation and detection methods may also play a role. PMID- 10803375 TI - The alkylated thiohydantoin method for C-terminal sequence analysis. AB - The alkylated-thiohydantoin method for C-terminal sequencing makes a significant improvement to the thiohydantoin method first described by Schlack and Kumpf. Prior to cleavage from the protein, the C-terminal thiohydantoin is alkylated, making it a better leaving group than the unmodified thiohydantoin. The C terminal alkylated-thiohydantoin can be cleaved from the protein under conditions that simultaneously form the next thiohydantoin. Combining cleavage and thiohydantoin formation in one step eliminates the need for activating the C terminal carboxyl group before every sequencing cycle and prevents detection of C termini formed by random cleavage of peptide bonds in the protein during the sequencing chemistry. The alkylated-thiohydantoin method includes the presequencing modification of cysteine and lysine and the automated modification of aspartic and glutamic acids, serine and threonine. Modifying the reactive side chain groups improves the ability to sequence through and detect these amino acids. The alkylated-thiohydantoin method can sequence through and detect 19 of the 20 genetically coded amino acids. Sequencing stops at proline residues. PMID- 10803376 TI - Ladder sequencing. AB - Ladder sequencing of polypeptides involves progressive N- or C-terminal amino acid truncation via chemical or enzymatic treatments. Peptide ladders are generated in which each component differs from the next by one residue. The ladder components are analyzed by mass spectrometry, and the amino acid sequence is deduced from the mass differences between consecutive fragments. Chemical procedures are common in N-terminal degradation, whereas proteolytic digestion is often used in C-terminal sequence analysis. Matrix-assisted laser desorption/ionization mass spectrometry is widespread for one-step readout of the peptide ladders and provides high sensitivity in combination with robustness and ease of use. The particular advantage of ladder sequencing in relation to other techniques for sequence analysis is the high data acquisition rate and the very good sample throughput that can be achieved. Multiple determinations are carried out within minutes at high sensitivity and low sample consumption. Several reports demonstrate analysis at the low picomole to femtomole level. PMID- 10803377 TI - New approaches for innovations in sensitive Edman sequence analysis by design of a wafer-based chip sequencer. AB - In the last few years the development of new mass spectrometric techniques enabled fast and sensitive protein analysis by the introduction of mass spectrometry (MS) fingerprinting and MS/MS sequencing. For these methods mixtures of peptide fragments of the proteins can be employed, whereas the Edman degradation method requests purified peptides. On the other hand, Edman sequencing has the advantages that interpretation of the data is more simple, extended sequences can be derived, and reliable sequence information on unknown proteins is possible. Hence, Edman chemistry as an alternative technique to MS is still valuable. But higher sensitivity of the sequencers is needed in order to meet modern demands, e.g. in proteomics research. We designed a wafer-based chip sequencer for protein and peptide sequencing in the femto- to attomole range. The main advantage of our new design is the complete integration of dead volume free valves together with reactor and converter and volume-measuring loops within one wafer-based system. In this system aggressive chemicals and solvents for the Edman degradation can be delivered in sub-microliter amounts, which allows a considerable shortening of the degradation cycles. Further, we developed sensitive maintenance and tightness tests to prove a precise and reproducible delivery of the chemicals and the reduction of drying times as compared with conventional sequencers. Real parallel processing of several samples can easily be implemented. The system is designed to serve future needs in protein research. PMID- 10803378 TI - Analysis of glycoprotein-derived glycopeptides. AB - Glycosylation of proteins represents one of the most important post-(co )translational events in view of the ubiquity of the phenomenon. In most cases, the covalently linked glycans are involved in the functioning of these biomolecules in biological systems. Detailed information on the carbohydrate moieties including monosaccharide composition, anomeric configurations, type of glycosidic linkages and attachment sites at the protein is indispensable in describing the ultimate structure of a specific glycoprotein. This chapter presents a general strategy for the structural characterization of glycoproteins/glycopeptides focussed on the glycan part. Some of the techniques commonly used, like enzyme treatments, separation methods, chemical analyses, mass spectrometry and nuclear magnetic resonance spectroscopy are briefly reviewed. PMID- 10803379 TI - Lipopeptide preparation and analysis. AB - Lipophilic peptides and proteins present specific problems during preparation and analysis which require the use of modified methodology. This chapter discusses some of the methods that have been employed in the isolation and structural studies of the pulmonary surfactant-associated proteins B and C (SP-B and SP-C), other proteins with lipid-like physicochemical properties, and the SP-B precursor. In particular, methods for separation and analysis of peptide/lipid mixtures, high-resolution separation of lipopeptides, analysis of fatty acylated peptides, and secondary and tertiary structure analysis of lipopeptides are discussed. PMID- 10803380 TI - Phosphopeptide analysis. AB - In this chapter we review the various methods available to the experimenter to analyse phosphorylated peptides. The initial steps in such an analysis involve the isolation of the phosphopeptides for analysis, and we outline the various current methods such as immobilised metal affinity chromatography, anti phosphoamino acid antibodies as well as HPLC (High Pressure Liquid Chromatography) and TLC (Thin Layer Chromatography). The isolated peptides can be analysed by chemical modification followed by Edman degradation or by mass spectrometry (MS). We focus on MS methods and give examples illustrating the selective detection and sequencing of phosphopeptides. PMID- 10803381 TI - Bioinformatics in protein analysis. AB - The chapter gives an overview of bioinformatic techniques of importance in protein analysis. These include database searches, sequence comparisons and structural predictions. Links to useful World Wide Web (WWW) pages are given in relation to each topic. Databases with biological information are reviewed with emphasis on databases for nucleotide sequences (EMBL, GenBank, DDBJ), genomes, amino acid sequences (Swissprot, PIR, TrEMBL, GenePept), and three-dimensional structures (PDB). Integrated user interfaces for databases (SRS and Entrez) are described. An introduction to databases of sequence patterns and protein families is also given (Prosite, Pfam, Blocks). Furthermore, the chapter describes the widespread methods for sequence comparisons, FASTA and BLAST, and the corresponding WWW services. The techniques involving multiple sequence alignments are also reviewed: alignment creation with the Clustal programs, phylogenetic tree calculation with the Clustal or Phylip packages and tree display using Drawtree, njplot or phylo_win. Finally, the chapter also treats the issue of structural prediction. Different methods for secondary structure predictions are described (Chou-Fasman, Garnier-Osguthorpe-Robson, Predator, PHD). Techniques for predicting membrane proteins, antigenic sites and postranslational modifications are also reviewed. PMID- 10803382 TI - [Recurrent dreams as migraine aura symptoms]. AB - Elementary geometric imagery seen in the visual aura of migraine can be experienced as incorporated into the content of a dream which precedes the awakening with a migraine headache. Furthermore, recurrent dreams featuring complex visual imagery, often terrifying nightmares, can occur as migraine aura symptoms. The said phenomena are illustrated by two original case reports and discussed against the background of a review of the literature. PMID- 10803383 TI - [Remembering, forgetting and depression. Time-conceptual implications of affective disorders]. AB - As a consequence of reflections about the relevance of remembering and forgetting in psychiatric disorders, aspects of the psychopathology of the dimension of time in phenomenological psychiatry are represented. From the perspectives of research on consciousness, cognition and neurobiology of the temporal lobes with special emphasis on memory processes, the impact of time in depressive illness is investigated. Based on Freud's concept of "Todestrieb" (death-instinct) and "Nirvana-Prinzip" (nirvana-principle) with regard to the possible impact on a theory of forgetting--based on Michael Theunissens time-philosophical concept, which demonstrates coping of "dominance of time" i.e. intrusion of the past into the presence and the failure of these coping-processes in depressive patients--it is hypothesized, that "nirvana-energy" is a major constituent of processes for forgetting. PMID- 10803384 TI - [Psychic pain as a symptom of depression. Phenomenology and neurobiology]. AB - In a review of the relevant literature the experience of pain in depression is emphasized. There is evidence for a central pain disturbance mediating the 'psychic pain' experience in severe major depressive episodes with a predominance of anxiousness and agitation. This phenomenological concept has not been considered as a specific construct in modern descriptive classification systems such as DSM-IV or ICD-10. The purpose of this article is to provide an overview of recent partially independent developments in depression and experimental pain research with emphasis on the neuroanatomy of the rostral limbic system and the medial pain system. The available evidence is consistent with the hypothesis that overlapping anatomic structures of the medial pain system are activated during the experience of both the psychic pain of depression and the motivational affective component of acute, tonic pain. This specific hypothesis can be tested with modern functional neuroimaging. PMID- 10803385 TI - [Negative delusional identity with schizophrenic psychoses. Case studies of phemonology and pathogenesis]. AB - With schizophrenics negative delusional identities constitute one way of psychotic alteration of self-identification. The main notion is of being a personification of evil. In a cross-cultural comparison study we found in the Austrian sample 13 patients with negative delusional identities. Our present study is based on detailed interviews and evaluations of medical records of this sample. Our aim was to draft a typology of delusional identities as a basic requirement for a phenomenology of the negative manifestations. Further investigative goals were the efforts of self-explanation undertaken by the patients with regard to their altered condition, the search for a pathogenetic transitional series and the functional value of the new identities. According to our estimation the basic mood on which negative delusional identities are founded is timid and dejected. Further basic requirements are a disturbed conscience of the ego and the concurrence of grandeur and guilt ideas. Half of our patients imagined to be reincarnations of negative biblical figures, three regarded themselves as possessed, two attributed their identities to heredity. Despite of diverse situative points of departure a common pathogenetic transitional series emerged for all patients. From a functional point of view a negative delusional identity seems to offer some kind of protection from further structural disintegration as well as relief from feelings of guilt--all that however at the price of structural deformations with dynamic depletion. PMID- 10803386 TI - [Subjective needs for care during the first year following hospital release by schizophrenic patients in the Dresden region]. AB - An initial cohort of 115 patients with chronic schizophrenia was studied during the period of care provided by community psychiatry programmes in the Dresden region one month and 12 months post hospital release using, among other instruments, the Berliner Bedurfnisinventar (Berlin Inventory of Care Needs). Thus a subjective need structure was identified over a one-year period, which is related primarily to autonomic basic social competence, a qualitatively sophisticated level of competence with regard to bonding and relationships, as well as (at least threatening) elements of social disintegration. Features characteristic of the course of disease (e.g. length of illness, changes in the psychopathological symptoms) and a factor of self-perceived disorder-related personal changes are the decisive predictors of how the extent of care needed will develop during the study period. Conclusions result from perceived deficits of care (e.g. with regard to employment/occupation). They pertain to a further content and institutionalized establishment of care-providing elements, which are not yet (currently) offered in a system of care that is being restructured along the lines of community psychiatry following German reunification. These conclusions fortify results of an analysis of normative needs for care and the current structure of community-based psychiatric care in the Dresden region. PMID- 10803388 TI - [The 97th meeting of the Japanese Society of Internal Medicine. Kyoto City, Japan. April 6-8, 2000. Abstracts]. PMID- 10803387 TI - [The carbamazepine hypersensitivity syndrome. Differential diagnosis and a representative case history]. AB - In psychiatry the efficacy of the anticonvulsant carbamazepine in the prophylaxis of bipolar affective disorders is very known and so this drug has been used for a long time. But one very important and dangerous side effect of carbamazepine must be emphasized: the "carbamazepine-hypersensitivity-syndrome", which will be presented in this article in a representative example. The allergic reaction can be difficult in differential diagnostic aspects: the clinical symptoms of this reaction can be very similar to viral infections; in the histological view aspects of a lymphoma can even be found. Because of the systemic reactions there are dramatic and very serious courses. So the psychiatrist has to remember the dramatic side effects and must inform the patients about them in using carbamazepine. PMID- 10803389 TI - [75th annual meeting of the Japanese Society for Tuberculosis. Osaka, Japan. April 17-19, 2000. Abstracts]. PMID- 10803390 TI - [The 59th annual meeting of the Japan Radiological Society. Yokohama, Japan. April 7-9, 2000. Abstracts]. PMID- 10803391 TI - [Regional meeting of the Japanese Society of Otolaryngology. Japan. 2000. Abstracts]. PMID- 10803392 TI - Transaxillary thoracoscopic sympathectomy experience in a hot climate: management of the dominant hand. AB - Primary palmar hyperhydrosis is a functionally and socially disabling problem of unknown etiology, affecting adolescents and young adults, especially in hot climates. Thoracoscopic sympathectomy is the most effective treatment for palmar hyperhydrosis. Postsympathectomy rebound hyperhydrosis may limit its success, especially in hot climates. The aim of this study is to report experience with thoracoscopic sympathectomy in a hot climate, managing the dominant hand (unilateral), followed by the other hand at a later date, based on the patient's choice. One hundred twenty patients were operated on during a 3 year period. The mean operative time was 25 minutes. The procedure was successfully completed in 169 operated limbs and was abandoned in one limb because of severe pleural adhesions. The procedure was done for the dominant hand (unilateral) in 120 patients. Fifty patients returned for contralateral thoracoscopic sympathectomy. There were 18 postoperative complications. Most of the patients (95%) were discharged after an overnight stay. The early observed cure rate was high (97%). During the mean follow-up period of 300 days, there was no recurrence of the original symptoms, except for one patient in whom the nerve of Kuntz was found and diathermized on the second thoracoscopy with symptomatic relief. Rebound hyperhydrosis occurred in 40 patients (33% of the total; 21% in the unilateral group and 42% in the bilateral group). In conclusion, it seems that transaxillary endoscopic sympathectomy of the dominant hand is an alternative method of treatment for patients with hyperhydrosis. Managing the dominant hand first and giving the patient the chance to observe the severity of the rebound hyperhydrosis may facilitate the decision for contralateral sympathectomy. PMID- 10803393 TI - The effect of dividing the sphincter of Oddi at endoscopic sphincterotomy on the filling and emptying aspects of function of the gallbladder. AB - The effect of dividing the sphincter of Oddi at endoscopic sphincterotomy (EST) on the filling and emptying aspects of gallbladder function has not been definitely established in humans. This prospective study is designed to examine the effects of EST on gallbladder emptying. In 13 patients (8 men and 5 women; mean age, 60.54 years +/- 2.14; range, 45-75 years), postprandial gallbladder emptying was measured ultrasonographically before and after EST (within 1-4 days). The fasting volumes after 8 hours of fasting, at times 0 min and before the test meal was given, residual volume (the smallest postprandial volume), gallbladder ejection fraction (EF), and total ejection volume, at 10, 20, 30, 40, 50, 60, 70, 80, and 90 min after the test meal, were studied. The fasting gallbladder volume and the residual volume tended to decrease after EST, but the differences were not significant (40.21 +/- 10.79 mL and 35.48 +/- 11.21 mL, 17.79 +/- 4.83 mL, and 13.10 +/- 4.83 mL, respectively; p > 0.05). Maximum EF was found to be 19.72% at 40 min and increased to 28.62% at 70 min after EST. Although the difference was not statistically relevant, a trend of improvement was evident after EST. The ejected volumes after EST have depicted a trend toward increase, without reaching to any statistical significance (p > 0.05). Our results demonstrate at least no adverse effects of EST and further support some positive effects on gallbladder kinetics. PMID- 10803394 TI - General anesthesia mode does not influence endocrine or immunologic profile after open or laparoscopic cholecystectomy. AB - To verify the impact on stress response and the influence of anesthesia on endocrine/immunologic changes, we have investigated the plasma level of norepinephrine, cortisol, TNFalpha, and IL-6 in 46 patients scheduled for laparotomy and laparoscopic cholecystectomy at 2, 6, 12, and 24 h after the operation. Among subjects who underwent open approach, 9 received fentanyl anesthesia and 13 received isoflurane anesthesia. In the laparoscopy group, 14 patients were given fentanyl anesthesia and 10 were given isoflurane anesthesia. The results obtained confirmed that laparoscopic cholecystectomy is associated with a lesser immunoendocrine response, and the two anesthesia models do not interfere with plasma changes of the assessed hormones and cytokines. PMID- 10803395 TI - Outpatient laparoscopic cholecystectomy: patient and nursing perspective. AB - Outpatient laparoscopic cholecystectomy has become nearly mandatory in the current health care economic environment. This study determined the opinions of patients and their nurses regarding disability and pain after laparoscopic cholecystectomy. Using a 14-point questionnaire, 32 consecutive patients who underwent inpatient LC were surveyed. The nurses caring for these patients were surveyed with a nine-point questionnaire. Seventy-one percent of patients believed that they could not have been discharged the day of surgery; the nurses reported that 81% of the patients could not have gone home. Nausea and vomiting were common. Pain was rated as 9 or 10 in 19% of patients. Nurses reported that 56% of postoperative patients were not receptive to discharge teaching. Morning (16) versus afternoon (16) procedures were not different. Successful programs of outpatient LC must: 1) ensure adequate home support because patient capability will be limited, 2) optimize pain control, 3) minimize nausea and vomiting, and 4) deliver patient education preoperatively. PMID- 10803396 TI - The effect of carbon dioxide pneumoperitoneum on the portal hemodynamics in a portal-hypertensive rat model. AB - As use of laparoscopic surgery grows, various complications with pneumoperitoneum have been reported. However, there are no reports on the relation between the carbon dioxide (Co2) pneumoperitoneum and portal hemodynamics. The purpose of this study was to determine how the pneumoperitoneum affects the portal hypertensive state. Portal-hypertensive rats, control rats, and sham operation rats were studied. The mean arterial pressure, portal venous pressure, central venous pressure, red blood flow volume of the viscera, and liver and renal function were investigated. The findings of this study suggest that Co2 pneumoperitoneum at less than 10 mm Hg does not present any undue risk to the portal hemodynamics in portal-hypertensive rats; however, more attention should be paid to the IAP in PHT patients undergoing surgery. PMID- 10803397 TI - Incisional hernia and fascial defect following laparoscopic surgery. AB - Complications involving the abdominal wall, particularly incisional hernias, were not expected when laparoscopic procedures were first introduced. With the increasing number of laparoscopies in abdominal surgery, more incisional hernias are observed. The authors report 13 cases of umbilical incisional hernia, which occurred late after laparoscopic cholecystectomy, and one case of omental procidentia through a lateral port, which occurred early after laparoscopic hernia repair with the transabdominal preperitoneal technique. There are 4 men and 10 women (mean age, 59.8 years; range, 40-74 years). Between March 1991 and December 1997, a total of 1,287 patients underwent laparoscopic operations at the Surgical Department of the Gradenigo Hospital in Turin, Italy. Incisional hernia incidence is 1%. Risk factors, such as chronic bronchitis or weight increase, which give rise to endoabdominal pressure, are present in some cases. Malnutrition may have a major role in many cases. Calculi larger than 15 mm are also seen frequently. Postlaparoscopy incisional hernia is generally a minor complication--only once did its occurrence cause a strangulated hernia. All precautions, including fascial suturing, must be taken to reduce the 1% incidence of postoperative incisional hernias. PMID- 10803398 TI - Video-assisted transsternal radical esophagectomy: three-field lymphadenectomy without thoracotomy for esophageal cancer. AB - To reduce the invasiveness of radical esophagectomy, we developed a new approach: video-assisted transsternal radical esophagectomy (VATRE). This article presents the operative techniques and our initial results. In our new procedure, cervical U-shaped and longitudinal sternoabdominal incisions are made, and a complete midline sternotomy is carried out. Lymph node clearance from the neck to the upper mediastinum and from the lower mediastinum to the upper abdomen is performed under direct vision. In the middle mediastinum, a video-assisted technique is used to dissect the lymph nodes. After esophageal resection and three-field lymphadenectomy, reconstruction is performed. One-lung ventilation is unnecessary. We have performed this procedure in two cases. These patients had no major complications and recovered more rapidly than patients undergoing conventional transthoracic esophagectomy. Our initial experience shows that VATRE is a technically feasible and less invasive procedure for cancer surgery, and it enables us to easily perform three-field lymphadenectomy. PMID- 10803399 TI - Laparoscopic transabdominal repair of hernia of Morgagni-Larrey. AB - A patient with symptomatic gall stones was found to have a hernia of Morgagni. The patient complained of upper abdominal symptoms along with heaviness in the chest and mild dyspnea. A complete diagnosis was possible with a chest X ray and a CT scan, which revealed a right-sided Morgagni hernia containing omentum and some bowel loops. It was decided to laparoscopically deal with both lesions at the same sitting. Initially, a laparoscopic cholecystectomy was accomplished. The hernial contents were then reduced and an 8 cm x 5 cm defect was closed with a tailored mesh sutured in place with a hernia stapler. Follow up after one month showed an asymptomatic patient confirmed by CT scan. Morgagni hernia is eminently treatable laparoscopically and must be considered as a first line approach to this problem. It can safely be combined with other laparoscopic procedures. PMID- 10803400 TI - Laparoscopic treatment of appendico-cecal intussusception. AB - Laparoscopy is a therapeutic possibility in pediatric surgery. A case of appendiceal intussusception treated laparoscopically in a 27-month-old girl is presented. The postoperative course was normal. PMID- 10803401 TI - Laparoscopic diagnosis and management of malignant ascites. AB - Malignancy is the second most common cause of ascites. Tissue diagnosis is often difficult because the cytology of ascitic fluid is positive in only 57% of cases. Peritoneovenous shunting is often used as palliation in such patients and has proven superior to nonoperative management for some patients. We present three cases of malignant ascites with negative cytologies managed by using laparoscopic biopsies to confirm intraperitoneal cancer and assist in the placement of a peritoneovenous shunt. Results suggest that exploratory laparoscopy and shunt placement is a valuable procedure in these patients with a limited life expectancy and is preferable to open laparotomy. PMID- 10803402 TI - Two-millimeter minisite mini-laparoscopy for rescue of dysfunctional continuous ambulatory peritoneal dialysis catheters. AB - In addition to peritonitis, mechanical outflow obstruction is the most common complication of continuous ambulatory peritoneal dialysis. If conservative methods are unsuccessful, the insufficient catheter must be placed in the right position or freed surgically. Currently, laparoscopy appears to be the method of choice in these cases. "Standard" laparoscopy using 10- and 5-mm ports can lead to dialysate wound leakage. Two-mm mini-laparoscopy was employed in three cases with good results. In two patients, catheters are still patent (after 19 and 23 months), and in one patient, the catheter had to be replaced after 5 weeks. No postoperative complications occurred. In some cases of catheter dislodgement or wrapping, mini-laparoscopy using a 2-mm MiniSite device can replace open surgery and even "standard" laparoscopy. PMID- 10803403 TI - Laparoscopic resection of an omental cyst with pedicle torsion. AB - Omental cysts are the least-common variety of all types of intra-abdominal cystic lesions. In the past, transabdominal laparotomy with excision of the cyst was the treatment of choice. With the advent of laparoscopic surgery, it has become possible to resect the cyst without the need for a large incision in the abdomen. We report a case of a 15-year-old girl who underwent diagnostic laparoscopy for recurring abdominal pain of 2 years' duration. The procedure revealed a huge cyst lying above the omentum with its pedicle rising from the greater curvature of the stomach. The pedicle was noted to have twisted eight times in a clockwise direction. The cyst was resected by laparoscopic means using three trocars. The postoperative course was uneventful. As presented in this case, we believe that a laparoscopic approach is an attractive alternative for the management of omental cystic lesions. PMID- 10803404 TI - A mouse model for breast cancer induced by amplification and overexpression of the neu promoter and transgene. AB - BACKGROUND: ErbB-2 is a critical oncogenic marker in human breast cancer. Its appearance correlates with poor prognosis and it is, therefore, an important target for physiologic investigation and therapeutic intervention. With this in mind, we have created and characterized two mouse breast cancer models that express rat wild type neu, the homologue of ErbB-2, and rat mutant neu under the control of the normal mouse neu promoter. These models in which the copy number of the neu gene is moderately amplified should more closely parallel the expression pattern of ErbB-2 seen in some cases of human breast cancer. MATERIALS AND METHODS: Transgenic mouse models were constructed by injecting one of the two pronuclei of a fertilized FVB/n egg and implanting it into a pseudopregnant Swiss /Webster mouse. Tissue expression was analyzed through the use of reverse transcription polymerase chain reaction and mammary histopathology examined by fixing, staining and mounting of the entire gland. RESULTS: In the former wild type model, we show that low level, long term expression of neu leads to abnormal lobuloalveolar development in virginal glands and incomplete regression in multiparous glands. Malignant foci form following multiple rounds of pregnancy and regression. In the latter model, a similarly directed transgene carrying the constitutively activated, mutant form of the rat neu gene, a stronger but similar phenotype is displayed. CONCLUSION: Evidently minor perturbations in amplified neu expression are sufficient to alter mammary development and induce malignant transformation. PMID- 10803405 TI - Activator protein-1 mediates induced but not basal epidermal growth factor receptor gene expression. AB - BACKGROUND: The epidermal growth factor receptor (EGFR) is expressed at different levels in many cell types and found overexpressed in many cancers. EGFR expression is increased or decreased in response to extracellular stimuli. We examined the effect of increased c-Jun expression on EGFR promoter activity. MATERIALS AND METHODS: We used DNAse I foot-printing analysis to determine the binding of activator protein 1 (AP-1) to the promoter region. We also used cotransfection experiments and western blotting analysis to determine the effect of AP-1 family members on EGFR expression. RESULTS: AP-1 was able to bind to at least seven sites in the EGFR promoter region. Cotransfection of MCF-7 cells with a c-Jun expression vector and the EGFR promoter reporter resulted in a 7-fold increase in promoter activity. JunB, but not c-fos, also enhanced the EGFR promoter activity. An A-Fos-dominant negative shown to inhibit Jun-dependent transactivation was able to prevent c-Jun induction of the promoter activity, but only slightly decreased the basal activity of the promoter. Furthermore, the A Fos dominant negative was able to inhibit phorbol ester induction of the EGFR promoter. Examination of EGFR expression of MCF-7 stable cell lines that overexpress c-Jun revealed an increase in EGFR expression. Additionally, a cisplatin-resistant cell line, A2780/CP70, which has an increase in AP-1 activity compared with the parental cell line, A2780, was found to have an increase in EGFR level. CONCLUSIONS: These results indicate that AP-1 can act to increase the expression of EGFR and may play a role in upregulation of EGFR in cancer cells. PMID- 10803406 TI - Prognostic value of a CCR5 defective allele in pediatric HIV-1 infection. AB - BACKGROUND: A deletion of 32 base pairs in the CCR5 gene (delta32 CCR5) has been linked to resistance to HIV-1 infection in exposed adults and to the delay of disease progression in infected adults. MATERIALS AND METHODS: To determine the role of delta32 CCR5 in disease progression of HIV-1 infected children born to seropositive mothers, we studied a polymerase chain reaction in 301 HIV-1 infected, 262 HIV-1 exposed-uninfected and 47 HIV-1 unexposed-uninfected children of Spanish and Italian origin. Infected children were further divided into two groups according to their rate of HIV-1 disease progression: rapid progressors who developed severe clinical and/or immunological conditions within the second year of life, and delayed progressors with any other evolution of disease. Among the latter were the long-term, non-progressors (LTNP) who presented with mild or no symptoms of HIV-1 infection above 8 years of age. Viral phenotype was studied for 45 delayed progressors. RESULTS: No correlation was found between delta32 CCR5 and mother-to-child transmission of HIV-1. However, the frequency of the deletion was substantially higher in LTNP, compared with delayed (p = 0.019) and rapid progressors (p = 0.0003). In children carrying the delta32 CCRS mutation, the presence of MT-2 tropic virus isolate was associated with a severe immune suppression (p = 0.028); whereas, the presence of MT-2 negative viruses correlated with LTNP (p = 0.010). CONCLUSIONS: Given the rapidity and simplicity of the assay, the delta32 CCR5 mutation may be a useful predictive marker to identify children with delayed disease progression who, consequently, may not require immediate antiretroviral treatment. PMID- 10803407 TI - Activation of platelet-activating factor (PAF) receptor stimulates nitric oxide (NO) release via protein kinase C-alpha in HEC-1B human endometrial epithelial cell line. AB - BACKGROUND: Impairment of the fertility in the platelet-activating factor (PAF) receptor transgenic female mice suggests changes in PAF functions can influence uterine receptivity. We hypothesized that vasodilatory actions of PAF in the uterus was exerted by PAF-mediated nitric oxide (NO) release via activation of isoenzyme-specific protein kinase C (PKC). MATERIALS AND METHODS: Inducible and endothelial NOS was shown by Reverse transcription polymerase chain reaction RT PCR in cDNA synthesized from RNA extract of proliferative and secretory endometrium as well endometrial epithelial cell lines HEC-1B. The effect of WEB2170, N(G)-monomethyl-L-arginine (L-NMMA) and Ro31-8220 on PAF mediated NO release by HEC-1B cell was determined. PAF induced translocation of PKCalpha in HEC-1B cell and its antagonist effect by Ro 31-8220 was studied by Western immunoblot analysis. PKC isoenzyme regulated by PAF was determined in HEC-1B cell lysate by immunoprecipitation. RESULTS: PAF-evoked a rapid and concentration dependent biphasic increase in total NO in human HEC-1B endometrial epithelial cell line [as measured by a Sievers NOA 280A NO Chemiluminescent Analyser.] This increase in NO release was attenuated by the PAF receptor antagonist, WEB2170. Inhibition of NO synthesis by N(G)-monomethyl-L-arginine produced marked dose dependent attenuation of PAF-mediated NO release, indicating nitric oxide synthase (NOS) activation. PAF-mediated NO release was also inhibited by the PKC inhibitor Ro 31-8220 and by the removal of extracellular calcium, suggesting a dependency on PKC and calcium, respectively. RT-PCR analysis showed expression of inducible NOS and endothelial NOS in human endometrium, myometrium and HEC-1B cells. Western immunoblot analysis showed PKCalpha, betaII and iota were the principal isozymes present in the HEC-1B cell line and normal endometrium, suggesting that both HEC-1B cells and normal endometrium have similar PKC isozymes. PAF induced the translocation of both PKCalpha and PKCiota within the time frame of NO release. The translocation of PKCalpha, but not PKCiota, was susceptible to inhibition by Ro 31-8220 that also inhibited PAF-evoked NO release, suggesting that PKCalpha is the principal isozyme involved in this process and that eNOS may be a substrate for PKCalpha. Kinase assays performed using immunoprecipitated PKCalpha showed that PAF (1 nM) activated PKCalpha that was inhibited by co-incubation with Ro31-8220 and Ca(2+)-free medium. CONCLUSIONS: This study demonstrates that PAF-stimulated NO release via PKCalpha in epithelial cells might regulate endometrial functions such as implantation and menstruation. PMID- 10803408 TI - mdm2 mRNA level is a prognostic factor in soft tissue sarcoma. AB - BACKGROUND: The oncogenic properties of murine double minute-2 (mdm2) protein over-expression, which mostly results from the interaction with the tumor suppressor p53, are well described and their negative impacts on the prognosis of affected patients is well characterized. However, clinical relevance of mdm2 mRNA expression is poorly investigated. MATERIALS AND METHODS: In this study, 65 soft tissue sarcoma (STS) samples were analyzed for mdm2 mRNA expression by a quantitative reverse transcription polymerase chain reaction (RT-PCR) approach using available validated ready-to-use assays based on the TaqMan technology (PE Applied Biosystems, Weiterstadt, Germany). Mdm2 data were correlated to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression calculated from the same sample. RESULTS: For patients with a mdm2/GAPDH mRNA ratio below 50 zmol/amol the survival was strikingly reduced in comparison to patients with a ratio of > or =50 (p = 0.0241). Multivariate Cox analysis showed that the difference in prognosis for patients with tumor stage 2 and 3 became even more pronounced between patients with a ratio of <50 zmol/amol and patients with a ratio of > or =50 (p = 0.0041; RR = 5.6). To test if the group with an mdm2 mRNA expression > or =50 is homogenous concerning the prognosis, the group was divided into three subgroups with values of 50 to <100, 100 to <500 and > or =500. The subgroup with values of 100 to <500 showed the best prognosis (p = 0.0164); whereas, the one with values of 50 to <100 showed the worst prognosis in this group and, in between, was the one with values of > or =500. After omitting patients of stage 1 and 4, the subgroup with values of 100 to <500 showed an even more striking best prognosis (p = 0.0015); the other subgroups remained in the same sequence. The risk of tumor-related death over 5 years was most conspicuous in patients with mdm2 mRNA expression <50 than in those with ratios of 100 to <500 displaying a 13.3-fold higher risk. In a comparison between mdm2 mRNA levels and P53 protein expression or p53 mutational status, no relationship was found. CONCLUSIONS: In our study, the mdm2 mRNA level appears to be an independent prognostic factor for STS patients, marking its role in STS genesis and as a potential factor for gene therapeutical approaches. PMID- 10803409 TI - Functional imaging of brain areas involved in the processing of coherent and incoherent wide field-of-view visual motion. AB - The brain areas involved in processing wide field-of-view (FOV) coherent and incoherent visual stimuli were studied using positron emission tomography (PET). The brains of nine subjects were scanned as they viewed texture patterns moving in the roll plane. Five visual conditions were used: (1) coherent clockwise (CW) wide-FOV (>100 degrees) roll motion; (2) coherent counter-clockwise (CCW) wide FOV roll motion; (3) wide-FOV incoherent motion; (4) CCW motion confined to the central visual field (approximately 55 degrees); and (5) a stationary control texture. The region most activated by the coherent-motion stimulus relative to the static one was the medial-occipital cortex, whereas both the medial- and lateral-occipital cortices were activated by incoherent motion relative to a static texture. Portions of the retroinsular parietal-temporal cortex, superior insula, putamen, and vestibulocerebellum responded specifically to the coherence of the stimulus, whereas a widespread lateralized activation was observed upon subtracting the CW scans from the CCW scans. The results indicate separate neural regions for processing wide-FOV motion versus stimulus coherence. PMID- 10803410 TI - Saccular and utricular inputs to single vestibular neurons in cats. AB - Saccular and utricular organs are essential for postural stability and gaze control. Although saccular and utricular inputs are known to terminate on vestibular neurons, few previous studies have precisely elucidated the origin of these inputs. We investigated the saccular and utricular inputs to single vestibular neurons in whole vestibular nuclei of decerebrated cats. Postsynaptic potentials were recorded from vestibular neurons after electrical stimulation of the saccular and utricular nerves. Ascending and descending axonal projections were examined by stimulating the oculomotor/trochlear nuclei and the cervical segment of the spinal cord, respectively. After each experiment, locations of recorded neurons were identified. The recorded neurons (140) were classified into vestibulo-spinal (79), vestibulo-oculo-spinal (9), and vestibulo-ocular (3) neurons based on antidromic responses; 49 other vestibular neurons were unidentified. The majority of recorded neurons were mainly located in the lateral vestibular nucleus. Most of the otolith-activated vestibular nuclei neurons seemed to participate in vestibulospinal reflexes. Of the total 140 neurons recorded, approximately one third (51) received saccular and utricular inputs (convergent neurons). The properties of these 51 convergent neurons were further investigated. Most (33/51) received excitatory postsynaptic potentials (EPSPs) after saccular and utricular nerve stimulation. These results implied that most of the convergent neurons in this study additively coded mixed information for vertical and horizontal linear acceleration. Based on the latencies of convergent neurons, we found that an early integration process for vertical and horizontal linear acceleration existed at the second-order level. PMID- 10803411 TI - Functions of the nucleus of the optic tract (NOT). I. Adaptation of the gain of the horizontal vestibulo-ocular reflex. AB - We studied the role of the nucleus of the optic tract (NOT) in adapting the gain of the angular vestibulo-ocular reflex (aVOR) in rhesus and cynomolgus monkeys using lesions and temporary inactivation with muscimol. The aVOR gain was adaptively reduced by forced sinusoidal rotation (0.25 Hz, 60 degrees/s) in a self-stationary visual surround, i.e., a visual surround that moved with the subject, or by wearing x0.5 reducing lenses during natural head movements. The aVOR gains dropped by 20-30% after 2 h and by about 30% after 4 h. Muscimol injections caused a loss of adaptation of contraversive-eye velocities induced by the aVOR, and their gains promptly returned to or above preadapted levels. The gains of the adapted ipsiversive and vertical eye velocities produced by the aVOR were unaffected by muscimol injections. Lesions of NOT significantly reduced or abolished the animals' ability to adapt the gain of contraversive aVOR-induced eye velocities, and the monkeys were unable to suppress these contraversive-eye velocities in a self-stationary surround. The lesions did not affect ipsiversive aVOR-induced eye velocities, and the animals were still able to suppress them. Lesions of NOT also affected the unadapted or "default" aVOR gains. After unilateral NOT lesions, gains of ipsiversive aVOR-induced eye velocity were reduced, while gains of contraversive aVOR-induced eye velocity were either unaffected or slightly increased. Consistent with this, muscimol injections into the NOT of unadapted monkeys slightly reduced the gains of ipsiversive and increased the gains of contraversive-eye velocities by about 8-10%. We conclude that each NOT processes ipsiversive retinal-slip information about visual surround movement relative to the head induced by the aVOR. In the presence of visual surround movement, the retinal-slip signal is suppressed, leading to adaptive changes in the gain of aVOR-induced contraversive horizontal eye velocities. NOT also has a role in controlling and maintaining the current state of the aVOR gains. Thus, it plays a unique role in producing and supporting adaptation of the gain of the horizontal aVOR that is likely to be important for stabilizing gaze during head movement. Pathways through the inferior olive are presumably important for this adaptation. PMID- 10803413 TI - Vestibular catch-up saccades in labyrinthine deficiency. AB - During rapid head rotations, saccades ipsiversive with compensatory vestibulo ocular reflex (VOR) slow phases may augment the deficient VOR and assist gaze stabilization in space. The present experiments compared these vestibular catch up saccades (VCUSs) with visually and memory-guided saccades. To characterize VCUSs and their relationship to deficiency of the initial VOR, we delivered random, whole-body transients of 1000 and 2800 degrees/s2 peak yaw acceleration around four different eccentric vertical axes in eight unilaterally and one bilaterally vestibulopathic subjects, as well as nine age-matched normal subjects. Eye and head movements were sampled at 1200 Hz using magnetic search coils. Subjects fixed targets at either 500 or 15 cm distance immediately before unpredictable onset of rotation in darkness. Under all testing conditions, normal subjects exhibited only compensatory vestibular slow phases and occasional anticompensatory quick phases. This behavior was also typical of unilaterally vestibulopathic subjects rotated contralesionally. When rotated ipsilesionally, however, vestibulopathic subjects had deficient slow-phase VOR gain with prolonged latency, and six of the nine exhibited saccadic movements in the compensatory direction (VCUSs). Higher head accelerations preferentially evoked VCUSs, but there were no preferred combinations of target distances and eccentric rotation axes. Peak velocities and durations of VCUSs increased with saccade amplitude. The latency distribution for VCUSs peaked around 70 ms, substantially shorter than reported for either visually guided express saccades or vestibular memory contingent saccades. The latency of each VCUS was highly correlated with the gaze error prior to that VCUS. The amplitude of VCUSs was calibrated to gaze position error, such that VCUSs reduced gaze error by an average of 37%. Thus when VOR slow-phase responses cannot compensate fully for head rotation, vestibular gaze position error can nevertheless calibrate the programming of VCUSs to augment the deficient VOR, much like catch-up saccades substitute for deficient visual pursuit. PMID- 10803412 TI - Functions of the nucleus of the optic tract (NOT). II. Control of ocular pursuit. AB - Ocular pursuit in monkeys, elicited by sinusoidal and triangular (constant velocity) stimuli, was studied before and after lesions of the nucleus of the optic tract (NOT). Before NOT lesions, pursuit gains (eye velocity/target velocity) were close to unity for sinusoidal and constant-velocity stimuli at frequencies up to 1 Hz. In this range, retinal slip was less than 2 degrees. Electrode tracks made to identify the location of NOT caused deficits in ipsilateral pursuit, which later recovered. Small electrolytic lesions of NOT reduced ipsilateral pursuit gains to below 0.5 in all tested conditions. Pursuit was better, however, when the eyes moved from the contralateral side toward the center (centripetal pursuit) than from the center ipsilaterally (centrifugal pursuit), although the eyes remained in close proximity to the target with saccadic tracking. Effects of lesions on ipsilateral pursuit were not permanent, and pursuit gains had generally recovered to 60-80% of baseline after about 2 weeks. One animal had bilateral NOT lesions and lost pursuit for 4 days. Thereafter, it had a centrifugal pursuit deficit that lasted for more than 2 months. Vertical pursuit and visually guided saccades were not affected by the bilateral NOT lesions in this animal. We also compared effects of these and similar NOT lesions on optokinetic nystagmus (OKN) and optokinetic after nystagmus (OKAN). Correlation of functional deficits with NOT lesions from this and previous studies showed that rostral lesions of NOT in and around the pretectal olivary nucleus, which interrupted cortical input through the brachium of the superior colliculus (BSC), affected both smooth pursuit and OKN. In two animals in which it was tested, NOT lesions that caused a deficit in pursuit also decreased the rapid and slow components of OKN slow-phase velocity and affected OKAN. It was previously shown that slightly more caudal NOT lesions were more effective in altering gain adaptation of the angular vestibulo-ocular reflex (aVOR). The present findings suggest that cortical pathways through rostral NOT play an important role in maintenance of ipsilateral ocular pursuit. Since lesions that affected ocular pursuit had similar effects on ipsilateral OKN, processing for these two functions is probably closely linked in NOT, as it is elsewhere. PMID- 10803414 TI - Seeing or not seeing where your hands are. AB - Previous findings have demonstrated the existence of a visual peripersonal space centered on the hand in humans and its modulatory effects on tactile perception. A strong modulatory effect of vision on touch perception was found when a visual stimulus was presented near the hand. In contrast, when the visual stimulus was presented far from the hand, only a weak modulatory effect was found. The aim of the present study was to verify whether such cross-modal links between touch and vision in the peripersonal space centered on the hand could be mediated by proprioceptive signals specifying the current hand positions or if they directly reflect an interaction between two sensory modalities, i.e., vision and touch. To this aim, cross-modal effects were studied in two different experiments: one in which patients could see their hands and one in which vision of their hands was prevented. The results showed strong modulatory effects of vision on touch perception when the visual stimulus was presented near the seen hand and only mild effects when the vision of the hand was prevented. These findings are explained by referring to the activity of bimodal neurons in premotor and parietal cortex of macaque, which have tactile receptive fields on the hand, and corresponding visual receptive fields in the space immediately adjacent to the tactile fields. One important feature of these bimodal neurons is that their responsiveness to visual stimuli delivered near the body part is reduced or even extinguished when the view of the body part is prevented. This implies that, at least for the hand, the vision of the hand is crucial for determining the spatial mapping between vision and touch that takes place in the peripersonal space. In contrast, the proprioceptive signals specifying the current hand position in space do not seem to be relevant in determining the cross-modal interaction between vision and touch. PMID- 10803415 TI - Vestibular perception of self-rotation in different postures: a comparison between sitting and standing subjects. AB - We investigated whether posture - either seated (S) or upright standing (O, orthostatic) - affects the vestibular perceptions of angular velocity (V) and displacement (D) in the horizontal plane. We also examined whether the two perceptions are equivalent, that is, whether perceived displacement can be viewed as the time integral of perceived velocity. Sinusoidal stimuli were delivered to subjects sitting on a Barany chair or standing on a turning platform. Frequencies ranged from 0.028 Hz to 0.45 Hz, peak-to-peak amplitudes from 11.3 degrees to 180 degrees, and peak velocities from 4 degrees/s to 64 degrees/s. Perceptions were measured by retrospective magnitude estimation in relation to a standard stimulus (STD) of 0.11 Hz, 45 degrees, 16 degrees/s. For D-estimates, two different moduli were assigned to the STD: Either "45 degrees" (allowing subjects to use the familiar degree scale, which can easily be related to the body scheme) or "10" (which bears no relation to an accustomed scale). For V-estimations the modulus was always "10" (there is no "natural" velocity scale). D-estimates exhibited only a marginal, non-significant dependence on posture (S larger than O); they were highly veridical (linear function of stimulus amplitude, gain close to 1) when subjects used the degree scale but had a reduced gain (approximately 0.76) with a modulus of 10. V-estimates, on the other hand, varied with posture (S significantly larger than O), particularly upon presentation of large stimuli; also, they deviated increasingly from veracity as stimulus magnitude increased (saturating function). Finally, posture had no effect upon the vestibular detection threshold. The frequency response of D-estimates, tested with stimuli of constant amplitude and varying frequency, was bimodal at low frequencies: stimuli were either not detected at all or were veridically estimated, on average (with a large scatter, though). The frequency response of V-estimates, tested with stimuli of constant peak velocity, exhibited a continuous increase with stimulation frequency. We conclude that published quantifications of vestibular self-motion perception, collected mostly with sitting subjects, are likely to be applicable also to the more natural situation of standing subjects provided they are based on displacement indications; in contrast, velocity indications appear to be modulated by posture. The different susceptibility of displacement and velocity estimates to posture and their incongruent frequency characteristics suggest that perceived displacement does not, or does not always, equal the time integral of perceived velocity. The persistence of nearly veridical displacement estimates at low frequencies suggests the intervention of cognitive processes. PMID- 10803416 TI - Motor cortical potentials precede long-latency EMG activity evoked by imposed displacements of the human wrist. AB - Rapid angular displacements of the wrist evoke cerebral potentials that precede the onset of the long-latency electromyographic (EMG) activity generated in muscles stretched by the displacement. The initial segment of the long-latency EMG activity (termed the M2 response) is thought to be mediated by a transcortical reflex. We used dipole source analysis to examine the source generators of the early components of the cerebral potentials and their relationship to the timing and magnitude of the M2 response. Subjects (n=10) were presented with instructions to either actively flex or extend the wrist in response to a torque motor-imposed extensor displacement or allow the wrist to be passively extended. Electroencephalographic (EEG) recordings were obtained from 32 scalp-surface electrodes, and EMG was recorded from the wrist flexors and extensors. For all three tasks, the M2 response was preceded by cerebral potentials that could be explained by a three-dipole model. One source generator localised to deep within the cerebrum, and the other two localised to the region of the contralateral sensorimotor cortex. We used the P20-N20 dipole evoked by electrical stimulation of the median nerve at the wrist, corresponding to synaptic activity within cortical area 3b, as a local spatial reference to examine the contributions of the pre- and postcentral cortex. This analysis showed that one of the sensorimotor dipoles was consistently located anterior to the P20-N20 dipole at a displacement (average 11.5 mm) appropriate for a generator originating within the deep layers of area 4 on the anterior bank of the central sulcus. The orientation of this dipole was also consistent with a precentral generator and not a reversal of the potentials generated by input to area 3b. The time course of the area-4 dipole moment (onset =35 ms, peak =54 ms) was appropriate to reflect synaptic activity onto corticospinal neurons whose descending volleys mediate the M2 response. Comparisons across tasks showed that the magnitude of the M2 was modulated with task instruction, being largest with active and smallest with passive resistance. In contrast, the magnitude of the early evoked potentials (up to 75 ms) did not grade across tasks. We interpret these results as suggesting that instruction-dependent modulation of the M2 response occurs downstream from inputs to the primary motor cortex. PMID- 10803417 TI - Connections of Purkinje cell axons of lobule X with vestibulospinal neurons projecting to the cervical cord in the rat. AB - Connections of Purkinje cell axons of lobule X (nodulus vermis) with vestibulospinal neurons have been demonstrated in the rat, by anterograde labeling of axons with biotinylated dextran (BD) injected into sublobule Xa and by retrograde labeling of neurons with cholera toxin subunit B (CTB) injected into cervical segments. Labeled terminals of Purkinje cell axons were numerous in the superior vestibular nucleus, the parvocellular (MVpc) and the caudal part (MVc) of the medial vestibular nucleus (MV), and group y. A limited number of labeled terminals were seen in the caudal part of the descending vestibular nucleus (DV). Occasional labeled terminals were seen in the lateral part of the lateral vestibular nucleus (LV) whereas few labeled terminals were seen in the magnocellular part of the MV (MVmc). Vestibulospinal neurons labeled from the C2 and C3 segments were seen bilaterally in the MVmc, MVpc, MVc, and DV, and ipsilaterally in the LV. CTB-labeled vestibulospinal neurons in contact with BD labeled terminals of Purkinje cell axons were identified in the lateral part of the MVpc, near the border between the MVpc and MVmc, or close to the dorsal acoustic stria, and in the middle part of the MVc at its rostral level. The present study suggests that Purkinje cells of lobule X regulate the output of cervical-projecting vestibulospinal neurons in the MVpc and MVc. PMID- 10803418 TI - Motor execution is necessary to memorize disparity. AB - Binocular saccades in response to briefly flashed, memorized disparate targets (different for the two eyes) become disconjugate following repeated trials. After 15 min of such training, the disconjugacy persists, even when the target to memorize is no longer disparate. This study examines the hypothesis that disparity memorization has a motor basis. We report here three experiments in which subjects were trained for 15-min periods. In experiment 1, subjects made no saccade after target presentation (static training); in experiment 2 subjects intended to make a saccade, but they actually made a saccade in only 10% or 20% of the trials; in experiment 3 subjects made anti-saccades. For all three experiments, the flashed target was disparate and the memory delay for each trial was 1 s. To examine the effects of learning for all three experiments, before and after training, we recorded memory-guided saccades to non-disparate targets (monocular viewing). Experiments 1 and 2 produced inconsistent (before/after training) changes in the disconjugacy of saccades. Thus, the disparity of potential saccade targets had no lasting effect on the disconjugacy of saccades if a saccade was not made. In contrast, the anti-saccades in experiment 3 developed a disconjugacy opposite to the disparity of the remembered target. These findings indicate that the execution of the saccade is necessary to memorize disparity of the target. PMID- 10803419 TI - Early corrective reactions of the leg to perturbations at the torso during walking in humans. AB - The contribution of afferent feedback to the regulation of locomotion in humans is not well understood. Animal experiments have suggested that loading of the leg during the stance phase may enhance the magnitude of extensor burst activity and delay the onset of swing phase. The aim of the present study was to determine whether transient loading of the leg at the end of stance would enhance extensor muscle activity and delay the onset of swing in walking humans. To test this hypothesis, we applied loads to the hips of subjects so that the load was applied along the long axis of the leg at the end of stance (down-back unsupported, DBU). This resulted in an unexpectedly complex reaction characterised by rapid co contraction of antagonist pairs of muscles around the ankle and knee and a prolongation of the stance phase. We speculated that the complexity of the reaction was, in part, due to a disturbance in equilibrium. To address this possibility, two additional perturbation paradigms were tested: (1) subjects held a rail during the loading paradigm (down-back supported, DBS), or (2) subjects received only a posteriorly directed perturbation of the hips, which added no additional load to the leg (backward unsupported, BU). As predicted, the DBS perturbation resulted in an enhancement of the ongoing soleus-muscle activity, and the unexpected tibialis anterior burst that was observed during the DBU paradigm was absent. Allowing the subjects to hold a rail substantially reduced the change in the timing of the step cycle observed in the DBU paradigm. The BU perturbation prolonged the stance phase duration and, as expected, resulted in a burst of activity in tibialis activity. This was usually accompanied by a reduction in the ongoing soleus activity. Two important conclusions are drawn from the present study. First, loading of the leg at the end of stance phase enhances the ongoing extensor-muscle activity. We suggest that afferent feedback responding to the increase load supported by the leg leads to rapid enhancement of the active extensor muscles to compensate for the increased load and prevent collapse of the leg. Interestingly, the duration of the stance phase was only marginally increased when loading was applied without a postural disturbance (DBS). Second, posterior perturbation of the centre of mass at the end of stance phase evokes an "automatic postural response" in tibialis anterior. Of particular interest, this evoked postural response can occur simultaneously with an enhanced activation of soleus. This indicates that the DBU perturbation employed in this study elicited two responses, one to prevent the collapse of the leg and the other to stabilise the centre of mass. PMID- 10803420 TI - Interactive effects of anoxia and general anesthesia during birth on the degree of CNS and systemic hypoxia produced in neonatal rats. AB - A model of global hypoxia during Caesarean-section (C-section) birth has been widely used to study long-term effects of birth hypoxia on central nervous system (CNS) function. However, the actual degree of CNS and systemic hypoxia produced by the birth insult in this model has never been characterised. Additionally, the way in which the dam is anaesthetised during the C-section procedure may impinge on the degree of hypoxia experienced by the neonate. This study examined how a period of global birth anoxia and isoflurane/N2O anaesthesia interact to affect measures of CNS and systemic hypoxia in neonatal rats born by C-section compared with control, vaginally born animals. A 10-min period of global anoxia just before birth increased blood lactate, a metabolic indicator of systemic hypoxia, increased brain lactate and decreased brain ATP to a similar extent in pups born by C-section from either decapitated, unanaesthetised dams or dams anaesthetised with 2.5% isoflurane. Thus, this model does produce systemic and CNS hypoxia in the neonate. Pups born by C-section with a higher concentration of isoflurane (3.5%), in the absence of added global anoxia, also showed reductions in brain ATP at birth. In addition, 10 min of global anoxia produced greater increases in blood lactate in pups born from dams anaesthetised with the higher concentration of isoflurane. Thus, the concentration of anaesthetic used in this model may affect the degree of CNS or systemic hypoxia experienced by the neonate. Compared with vaginal birth, pups born by C-section with 2.5% or 3.5% isoflurane (and no added global anoxia) showed decreased PO2 and pH, and increased pCO2 in systemic blood taken <30 s after birth. Exposure to global anoxia during C-section birth actually increased systemic PO2 at <30 s after birth, presumably due to ventilatory responses to hypoxemia and hypercapnia; this effect of anoxia was reduced in anaesthetised compared with unanaesthetised pups. Thus, global anoxia acts as a stimulus for rapid recovery of systemic PO2 at birth, and this stimulus is dampened by isoflurane/N2O anaesthesia. These results should aid in understanding how CNS and systemic hypoxia at birth contribute to long-term changes in brain biochemistry and behaviour in this model. PMID- 10803421 TI - A forty-two year voyage through vision research. AB - This presentation is an overview of my involvement in vision research and the factors and individuals that influenced my career in this field over the last 42 years. It also summarizes my research interests and contributions in the areas of aqueous humor dynamics, transport of various substances across blood-aqueous barrier and in the lens. The metabolism and function of glutathione in the lens and the development of tissue culture of human lens epithelium as a model system to study its role in lens and cataract formation are reviewed. PMID- 10803422 TI - Retinal vascular changes induced by the oxidative stress of alpha-tocopherol deficiency contrasted with diabetic microangiopathy. AB - It has been proposed that oxidative tissue damage is involved in the development of diabetic angiopathies. To evaluate this hypothesis, experiments were conducted to identify the retinal vessel changes induced by the oxidative stress related to alpha-tocopherol deficiency and examine possible similarities with the lesions characteristic of diabetic retinopathy. Twenty-one-day-old male Fisher 344 albino rats were divided randomly to receive a basal, chemically defined diet either with (adequate group) or without (deficient group) alpha-tocopherol. After 6 and 8 months, some rats (n = 3 per group) were killed and the eyes removed. In order to evaluate cell integrity and localization of lipofuscin-specific autofluorescence by light and fluorescence microscopy, some of the retinas were prepared for cryostat-sections while others were digested by elastase to isolate intact retinal vasculatures. After 8 and 14 months, the central retina of one eye per rat (n = 6 to 8 per group) was examined by electron microscopy for retinal capillary basement membrane (RCBM) thickening and other ultrastructural changes. At 6 and 8 months, the deficient rats exhibited extensive shortening and disarray of rod outer segments (ROS), marked loss of photoreceptor cells, and pronounced increases in the numbers of granules with lipofuscin-specific autofluorescence in the retinal pigment epithelium (RPE) and retinal vessels. At 14 months, the ultrastructure revealed that the damage to ROS involved disruption of membranes and that the capillary lipofuscin was contained mainly within the endothelial cells. Membrane remnants were found in the lipofuscin granules of both the RPE and retinal vessels. In addition, there was an increase in RCBM thickness (98.7 +/- 2.6 nm vs. 86.9 +/- 2.9 nm). RCBM thickening was the only finding common with diabetic retinopathy, and the thickening was 13.6%, significantly less than that reported in diabetic rat models with 8 and 14 months durations (34% and 53.1%, respectively). Capillary lipofuscin accumulation, which was prominent in the deficient rats, is not notable in diabetes. Both the moderate RCBM thickening and marked lipofuscin accumulations seen in alpha-tocopherol-deficient rats were similar to changes occurring in the aging process, though more pronounced. The spectrum of microangiopathies characteristic of diabetic retinopathy did not develop in alpha-tocopherol-deficient rats. These findings suggest that oxidative damage, though probably involved, is unlikely to play a predominant role in the development of diabetic retinal microangiopathies. PMID- 10803423 TI - Glutathione: a vital lens antioxidant. AB - The reducing compound glutathione (GSH) exists in an unusually high concentration in the lens where it functions as an essential antioxidant vital for maintenance of the tissue's transparency. In conjunction with an active glutathione redox cycle located in the lens epithelium and superficial cortex, GSH detoxifies potentially damaging oxidants such as H2O2 and dehydroascorbic acid. Recent studies have indicated an important hydroxyl radical-scavenging function for GSH in lens epithelial cells, independent of the cells' ability to detoxify H2O2. Depletion of GSH or inhibition of the redox cycle allows low levels of oxidant to damage lens epithelial targets such as Na/K-ATPase, certain cytoskeletal proteins and proteins associated with normal membrane permeability. The level of GSH in the nucleus of the lens is relatively low, particularly in the aging lens, and exactly how the compound travels from the epithelium to the central region of the organ is not known. Recently, a cortical/nuclear barrier to GSH migration in older human lenses was demonstrated by Sweeney et al. The relatively low ratio of GSH to protein -SH in the nucleus of the lens, combined with low activity of the glutathione redox cycle in this region, makes the nucleus especially vulnerable to oxidative stress, as has been demonstrated with use of in vivo experimental animal models such as hyperbaric oxygen, UVA light and the glutathione peroxidase knockout mouse. Effects observed in these models, which are currently being utilized to investigate the mechanism of formation of human senile nuclear cataract, include an increase in lens nuclear disulfide, damage to nuclear membranes and an increase in nuclear light scattering. A need exists for development of therapeutic agents to slow age-related loss of antioxidant activity in the nucleus of the human lens to delay the onset of cataract. PMID- 10803424 TI - Thiol regulation in the lens. AB - The high content of glutathione (GSH) in the lens is believed to protect the thiols in structural proteins and enzymes for proper biological functions. The lens has both biosynthetic and regenerating systems for GSH to maintain its large pool size (4-6 mM). However, we have observed that, in aging lenses or lenses under oxidative stress, the size of GSH pool is diminished; and some protein thiols are being S-thiolated by oxidized nonprotein thiols to form protein-thiol mixed disulfides, either as protein-S-S-glutathione (PSSG) or protein-S-S cysteine (PSSC). We have shown in an H2O2-induced cataract model that PSSG formation precedes a cascade of events starting with protein disulfide crosslinks, protein solubility loss, and eventual lens opacification. Recently, we discovered that this early oxidative damage in protein thiols could be spontaneously reversed in H2O2 pretreated lenses if the oxidant was removed in time. This dethiolation process is likely mediated through a redox regulating enzyme, thioltransferase (TTase), which has been discovered recently in the lens. To understand if the role of oxidative defense and repair is the physiological function of TTase in the lens, we cloned the TTase gene and purified the recombinant human lens TTase. Although TTase required GSH for its activity, TTase was far more efficient in dethiolating lens proteins than GSH alone. It favored PSSG over PSSC and dethiolated gamma-crystallin-S-S-G better than the alpha crystallin counterparts. Furthermore, TTase showed a remarkable resistance to oxidation (H2O2) in cultured rabbit lens epithelial cells when GSH peroxidase, GSH reductase, and glyceraldehyde-3-phosphate dehydrogenase were severely inactivated. We further showed that activity loss in those SH sensitive enzymes could be attributed to S-thiolation, but reactivation via dethiolation could be attributed to TTase. We conclude that TTase can regulate and repair the thiols in lens proteins and enzymes through its dethiolase activity, thus contributing to the maintenance of the function of the lens. PMID- 10803425 TI - Relative importance of aldose reductase versus nonenzymatic glycosylation on sugar cataract formation in diabetic rats. AB - The relative importance of sorbitol formation versus nonenzymatic glycosylation and advanced glycosylation end products (AGEs) on sugar cataract formation was examined in diabetic rats. Diabetes was experimentally induced in young, 50 g rats with streptozotocin, and aldose reductase inhibitors were administered in the diet for up to 8 weeks at concentrations of 0.06% for tolrestat or ponalrestat and 0.0125% for AL-1576. Cataract formation was monitored by hand held slit lamp for up to 11 weeks. Lens polyol levels were monitored by GLC, glycosylated protein levels were spectrophotometrically determined, and AGE products were estimated by fluorescence measurements and ELISA. Sugar cataract formation was observed in all untreated diabetic rats while cataract formation was inhibited in all diabetic rats treated with the AR inhibitors. Lens sorbitol levels were reduced in all ARI-treated rats. Glycosylated lens protein levels were elevated in the diabetic rats, and these levels were not significantly lower in the non-cataractous lenses from ARI-treated diabetic rats. Fluorescence measurements of the lens proteins revealed increased lens AGE levels in all diabetic rats, and these were slightly reduced in the aldose reductase inhibitor treated diabetics. With ELISA, immunoreactive AGEs were only detected in cataractous lenses from the untreated diabetic rats. Immunoreactive AGEs were not detected in the clear lenses of the aldose reductase inhibitor treated diabetics or in the non-diabetic controls. These results support the concept that sugar cataract formation is initiated by the aldose reductase catalyzed intracellular accumulation of polyols and that these sugar cataracts can be prevented through inhibition of aldose reductase. PMID- 10803426 TI - Lens UVA photobiology. AB - Although UVA radiation was considered to be benign as a factor that could contribute to cataract formation, the studies briefly summarized herein show that UVA may provide the more damaging radiant energy involved in the formation of human cataract. Of the UVA impinging onto the eye from sunlight, a sufficient dose does reach into the lens to enhance cataractous changes. The adverse physiological and biochemical effects on lens epithelial cells and their ability to differentiate into clear adult lenses include cell growth inhibition, cell membrane and cytoskeletal anomalies, and enzyme inactivation. Many of the damaging events are related to the excitation of sensitive UVA absorbing chromophores which generate singlet oxygen and free radicals. This process enhances lens oxidative stress. Some protection against this damage is provided by vitamin E and tea polyphenols. Thus, an imbalance between photooxidative stress, antioxidant protection, and repair processes would increase the potential to produce cataracts. PMID- 10803427 TI - Retinal capillary dilation: early diabetic-like retinopathy in the galactose-fed rat model. AB - The purpose of this study was to determine whether capillary dilation is one of the earliest structural changes in the diabetic-like retinopathy of the galactose fed rat model and thus may represent a stage where intervention treatment might still be effective. Weanling female Sprague-Dawley rats were randomized into 3 groups and fed Purina laboratory chow plus one of the following for 4 months: 50% starch (CONTROL); 50% D-galactose (Galactose); or 50% D-galactose with ARI-509 (25 mg/kg body wt/day) (Inhibitor). One eye from each of 5 rats per treatment group was processed for retinal vasculature wholemounts using elastase digestion, stained with a standard periodic-acid-Schiff reaction and counterstained with hematoxylin. Average capillary width, overall capillary density and total capillary length were measured, using computerized image analysis, within an arc shaped area (4.4 mm2) of each vasculature surrounding, but separated from, the optic disc margin by approximately 0.7 mm. Galactose rats exhibited significant (p < 0.001) increases in capillary width (Mean +/- SEM: 7.56 +/- 0.07 microm) and density (42.78 +/- 0.37%) compared with CONTROL rats (6.68 +/- 0.11 microm and 37.18 +/- 0.30%, respectively). These increases were prevented with inhibitor treatment (6.58 +/- 0.16 microm and 35.88 +/- 0.97%, respectively). Capillary length remained unchanged at 4 months ( CONTROL: 246.66 +/- 2.46 mm; Galactose: 250.75 +/- 1.26 mm; Inhibitor: 242.25 +/- 8.43 mm). Retinal capillary dilation, expressed as increased width and density, is one of the earliest detectable lesions in galactose-fed rats. In these rats, the lesion occurs as early as retinal capillary basement membrane thickening (RCBMT), one of the earliest reported changes in human diabetic retinopathy. Like RCBMT, capillary dilation can be prevented in rats with aldose reductase inhibitor treatment. Unlike RCBMT, capillary dilation could be clinically detectable and may be useful for the diagnosis of early retinopathy and for determining the timing of therapeutic intervention. PMID- 10803428 TI - Review: A case for corneal crystallins. AB - It is established that the diverse, multifunctional crystallins are responsible for the optical properties of the cellular, transparent lens of the complex eyes of vertebrates and invertebrates. Lens crystallins often differ among species and may be enzymes or stress proteins. I present here the idea that abundant water soluble enzymes and other proteins may also be used for cellular transparency in the epithelial cells and, possibly, stromal keratocytes of the cornea. Aldehyde dehydrogenases and transketolase are among the putative "corneal crystallins" in mammals, and gelsolin may be a corneal crystallin in the zebrafish. In invertebrates, the glutathione S-transferase-related S-crystallins of the lens appear to be used also as corneal crystallins in the squid, and an aldehyde dehydrogenase-related protein is the crystallin in the lens and, possibly, cornea of the scallop. The use of abundant, taxon-specific water-soluble proteins as crystallins for cellular transparency in the cornea would provide a new conceptual link between this tissue and the lens. PMID- 10803430 TI - Review: Oxidative stress and disease. AB - This communication briefly reviews aspects of oxidative stress and disease, particularly maturity onset cataract. The review considers a number of issues such as why lens and cataract research is important, what is oxidative stress, its relationship to disease and how does a tissue defend itself against such stress. Three diseases (chronic lung disease, diabetes and Alzheimer's disease) in which oxidative stress has differing roles are briefly discussed. The impact of oxidative stress upon the development of maturity onset cataract is considered and approaches are delineated which will establish the importance of such stress. PMID- 10803429 TI - Review: Age-related cataract: immunity and lens epithelium-derived growth factor (LEDGF). AB - This short review summarizes our recent work and relevant publications on autoimmunity and cataract. A complete review of this subject is beyond the scope of this paper. Age-related cataract (ARC) is the leading cause of world blindness. In spite of more than fifty years of basic and clinical research, there is no nonsurgical intervention to prevent or treat ARC, but there is a better understanding of the manifold complexities of this age-related condition. ARC is a multifactorial condition in which incidence and progress are modified by factors such as age, sex, radiation [visible, ultraviolet (UV), and X-ray], oxidation, physical trauma, diet, and medications. The lens contains at least three different cell types: central epithelial cells, dividing germinative epithelial cells, and fiber cells. The central epithelial cells covering the anterior axial part of the lens do not divide but survive throughout life. The bulk of the lens comprises anucleate fiber cells, differentiated germinative epithelial cells, which have undergone an apoptosis-like change "diffoptosis" to become elongated, crystallin-rich, organelle-deficient, cells. The epithelial cells and their active transport mechanisms maintain lens homeostasis and clarity. The survival mechanisms of the central lens epithelial cells (LECs) are unknown. In other cells, growth or survival factors, when present, enhance survival and, when absent or deficient, induce programmed cell death "apoptosis". Many developing mammalian cells produce signal proteins, or require signal proteins from other cells, to avoid apoptosis. Although much is known about the role of growth factors in the lens, less is known about how such signals are involved in the survival and death of LECs. We have hypothesized that LECs, like other mammalian cells, use signal proteins to regulate growth, survival, and apoptosis, and we have begun a search for such molecules. Furthermore, we have hypothesized that such factors, if found, may also be involved in the death of LECs, the consequent alteration of lens homeostasis and, eventually, certain types of ARC. PMID- 10803431 TI - The status of the genera Hysterolecithoides Yamaguti, 1934, Neotheletrum Gibson & Bray, 1979 and Machidatrema Leon-Regagnon, 1998 (Digenea: Hemiuroidea), including a description of M. leonae n. sp. from Australian waters. AB - A diagnosis is given for the lecithasterid genus Hysterolecithoides Yamaguti, 1934, which is now found to have two to six (possibly seven) vitelline masses. The species H. frontilatus (Manter, 1969) is returned to the genus, having been considered a member of the bunocotylid genus Neotheletrum by recent authors. It is redescribed from Siganus nebulosus, Moreton Bay, and S. doliatus, Lizard Island, Great Barrier Reef and New Caledonia, with emphasis on the presence of Juel's organ, a uterine seminal receptacle and the blind sac associated with the genital atrium. It differs from its congeners in the trajectory of the pars prostatica which recurves dorsally to the sinus-sac. Oligolecithoides Shen, 1982 is synonymised with Hysterolecithoides and O. trilobatus Shen, 1982 is synomised with H. epinepheli Yamaguti, 1934. Machidatrema Leon-Regagnon, 1998 is diagnosed, and found to be close to Hysterolecithoides, but differs in the lack of a blind sac projecting from the dorsal genital atrium, by its tandem testes, the coiling of the uterus between the testes and the ovary, and the ventral excretory pore. M. leonae n. sp. is described from Siganus fuscescens, S. lineatus, S. doliatus, S. corallinus, S. vulpinus and Scarus globiceps at Heron Island, Queensland. It differs from its closest congener, M. akeh, in the muscular and tegumental flap over the genital pore and details of the terminal genitalia. M. chilostoma (Machida, 1980) and M. kyphosi (Yamaguti, 1970) are redescribed from Kyphosus vaigiensis from Heron Island. Neotheletrum Gibson & Bray, 1979 is diagnosed: it differs from Hysterolecithoides in its confluent excretory arms, blind seminal receptacle (no Juel's organ) and uniformly tripartite vitellarium. A cladistic analysis suggests that M. chilostoma and M. kyphosi are not best accommodated in Machidatrema, that Machidatrema (sensu stricto) is monophyletic and that Hysterolecithoides is paraphyletic. Hysterolecithoides and Machidatrema are considered hysterolecithine lecithasterids, whilst Neotheletrum is retained as an opisthadenine bunocotylid. PMID- 10803432 TI - Two new species of Caryospora Leger, 1904 (Apicomplexa, Eimeriidae) from accipitrid raptors. AB - Two new species of Coccidia (Apicomplexa: Eimeriidae) are described from European accipitrid raptors (Falconiformes: Accipitridae). Oocysts of Carvospora aquilae n. sp. found in faeces of the gold eagle Aquila chrysaetos are subspherical to broad ellipsoidal and measure 43 (40-49) x 37.5 (34-39) microm. Polar granule, oocyst residuum and micropyle are absent. Each oocyst contains one spherical to subspherical slightly polygonal sporocyst measuring 23.8 (23-25) x 23.3 (22-25) microm. Stieda and substieda bodies are absent. The sporocyst residuum is composed of numerous small granules less than 0.5 microm in diameter dispersed randomly among the sporozoites. Sporulated oocysts of Carvospora circi n. sp. from faeces of the marsh harrier Circus aeruginosus are widely oval, measuring 24.5 (23-25) x 21.8 (21-24) microm. A polar granule, oocyst residuum and micropyle are absent. Each oocyst contains one spherical to subspherical sporocyst measuring 16.2 (15-17) x 15.6 (15-17) microm. A compact granular, spherical to subspherical sporocyst residuum, 10.4 (10-11) x 8.5 (7-9), was present in 76% of measured sporocysts. In 24% of sporocysts the granules of sporocyst residuum were scattered among the sporozoites. PMID- 10803433 TI - Boreascotia megavesicula n. g., n. sp. (Digenea: Hemiuridae: Lecithochiriinae) in the nototheniid fish Lepidonotothen macrophthalma (Norman) from the sub-Antarctic Atlantic. AB - Boreascotia megavesicula n. g., n. sp. is described from the intestine of the fish Lepidonotothen macrophthalma from the North Scotia Ridge, Sub-Antarctic, Atlantic Ocean. The new genus has an ecsoma, placing it in the family Hemiuridae, and a distinct prostatic vesicle, placing it in the subfamily Lecithochiriinae, but is distinct from the other members of the subfamily in having an elongate, tubular seminal vesicle, which reaches well into the hindbody and overlaps the anterior testis. PMID- 10803434 TI - Morphological and molecular analyses of the genera Peltidocotyle Diesing 1850 and Othinoscolex Woodland 1933, and a morphological study of Woodlandiella Freze, 1965 (Eucestoda, Proteocephalidea), parasites of South American siluriform fishes (Pimelodidae). AB - Morphological studies of type-species of the genera Peltidocotyle Diesing, 1850 and Othinoscolex Woodland, 1933 showed that these genera are synonyms. We therefore redescribe the type and fresh material of Peltidocotyle rugosa Diesing, 1850 and P. lenha (Woodland, 1933) [= Othinoscolex lenha Woodland, 1933], which appears to constitute two distinct species. They differ in the number and number of layers of testes, the diameter of the metascolex and the field occupied by the vitelline follicles. Phylogenetic analyses based on partial DNA sequences of 16S rDNA, 5.8S-ITS2 and 28S rDNA support these results. P. lenha and Woodlandiella myzofer (Woodland, 1933) are considered conspecific following a thorough morphological examination. With regard to host-parasite relationships, P. lenha has been reported from two host species, which is unusual as South American proteocephalideans generally display an oioxenous type of host-specificity. PMID- 10803435 TI - Novel isotrichid ciliates endosymbiotic in Australian macropodid marsupials. AB - Samples of foregut content were collected from 86 macropodid marsupials and examined for the presence of endosymbiotic ciliates. Four host species were examined: Macropus giganteus (eastern grey kangaroo), M. fuliginosus (western grey kangaroo), M. robustus (common wallaroo) and Thylogale billardierii (Tasmanian pademelon). Ciliate morphology was determined by microscopical examination of live and silver-impregnated specimens. Isotrichid ciliates were detected in 51 (59%) of the 86 animals examined. Five new species and one new genus are described. Three of the species belong to the genus Dasytricha Schuberg, 1888; D. dehorityi n. sp. from M. giganteus, D. dogieli n. sp. from M. robustus and D. mundayi n. sp. from T. billardierii. Bitricha n. g. is characterised by the possession of two fields of somatic ciliation, a transverse ventral and a longitudinal dorsal field. B. oblata n. sp. is described from M. giganteus and M. fuliginosus and B. tasmaniensis n. sp. is described from T. billardierii. The occurrence of isotrichid ciliates in both metatherian and eutherian mammals suggests that the family either evolved prior to the divergence of the mammalian lineages or switched hosts from one group to the other following host diversification. PMID- 10803437 TI - The cercaria of Galactosomum bearupi Pearson, 1973 (Trematoda: Heterophyidae) at Heron and Masthead Islands, Great Barrier Reef, Australia. AB - Magnacercous cercariae, all morphologically identical but with different coloured tails, were found naturally infecting the intertidal prosobranch gastropod Clypeomorus batillariaeformis Habe & Kosuge (Cerithiidae) at Heron and Masthead Islands. Several species of coral-dwelling fishes were exposed to magnacercous cercariae. Active ingestion by the fish, followed by the complete development of the metacercaria in the optic lobes, occurred in Pomacentrus molluccensis Bleeker, but not in Dascyllus aruanus (L.), where development stopped short after encystment. All cercariae turned out to belong to the same species, Galactosomum bearupi Pearson, 1973. Natural infections of G. bearupi metacercariae were found in eight species of fish at Heron Island: Pomacentrus molluccensis, P. wardi Whitley, P. bankanensis Bleeker, P. flavicauda Whitley, Stegastes cf. fasciolatus (Ogilby), Sillago maculata Quoy & Gaimard, S. cf. ciliata Cuvier and Crenimugil crenilabris (Forsskal). This represents the fourth account of the cercaria of a species of Galactosomum. PMID- 10803436 TI - Enzymatic polymorphism and phylogenetic relationships in Leishmania Ross, 1903 (Sarcomastigophora: Kinetoplastida): a case study in Colombia. AB - Leishmaniasis is widespread in Colombia and is found in 30 of 32 Departments. More than 200 infection zones have been reported from different regions, which vary from sea-level to an altitude of 2,300 m along the Atlantic Coast, Pacific coast, Amazon basin, Cauca and Magdalena valleys. We report 76 Leishmania stocks isolated from humans, dogs and phlebotomine hosts. Isoenzyme electrophoresis revealed 16 zymodemes, which could be divided into four phylogenetic complexes, i.e., L. braziliensis, L. amazonensis, L. guyanensis/panamensis and L. infantum. Three zymodemes became integrated into the subgenus Leishmania and the other zymodemes into the subgenus Viannia. Cutaneous infections were due to the L. braziliensis (9.2%) and L. guyanensis/panamensis (85.54%) complexes. Mucous secondary involvement was due to the L. braziliensis and L. guyanensis/panamensis complexes. In this work the specific status of L. (V.) guyanensis and L. (V.) panamensis is discussed. PMID- 10803438 TI - The implantable cardioverter-defibrillator. AB - Ventricular arrhythmias remain a major cause of cardiovascular mortality. Therapy for serious ventricular arrhythmias has evolved over the past decade, from treatment primarily with antiarrhythmic drugs to implanted devices. The implantable cardioverter-defibrillator (ICD) is the best therapy for patients who have experienced an episode of ventricular fibrillation not accompanied by an acute myocardial infarction or other transient or reversible cause. It is also superior therapy in patients with sustained ventricular tachycardia (VT) causing syncope or hemodynamic compromise. Controlled clinical trials have confirmed the utility of these devices. As primary prevention, the ICD is superior to conventional antiarrhythmic drug therapy in patients who have survived a myocardial infarction and who have spontaneous, nonsustained ventricular tachycardia, a low ejection fraction, inducible VT at electrophysiologic study, and whose VT is not suppressed by procainamide. The effect of the ICD on survival of other patient populations remains to be proven. The device is costly, but its price is generally accepted to be reasonable. The ICD has been a major advance in the treatment of ventricular arrhythmias. PMID- 10803439 TI - Seropositivity against Chlamydia pneumoniae in patients with coronary atherosclerosis. AB - BACKGROUND: Results of therapy in patients with unstable coronary syndromes with antibiotics directed against Chlamydia pneumoniae have been variable, perhaps due to the heterogeneity of patients in these trials. HYPOTHESIS: The aim of the present study was to correlate the severity of coronary artery disease (CAD) with seropositivity against C. pneumoniae prospectively. METHODS: We measured the frequency of seropositivity (IgG levels > or = 1/64 and IgA levels > or = 1/16 against Chlamydia pneumoniae) in 110 patients with CAD and in 49 controls. RESULTS: As expected, traditional CAD risk factors were seen more often in patients with CAD than in controls. Mean values of total cholesterol (184 +/- 52 and 166 +/- 44 mg/dl, respectively) and triglyceride (143 +/- 60 and 112 +/- 63 mg/dl, respectively) in serum were significantly higher in patients with CAD than in controls (both p < 0.04). There were no significant differences between the two groups in serum high-density lipoprotein cholesterol (34 +/- 13 and 32 +/- 14 mg/dl, respectively) and lipoprotein (a) (Lp(a):241 +/- 247 and 223 +/- 263 mg/l, respectively) levels. The rate of IgG seropositivity was 52% (28/54) in patients with stable CAD, 41% (23/56) in patients with unstable CAD, and 35% in controls (p = NS). The rate of IgA seropositivity was 25% (14/54) in patients with stable CAD, 12% (6/49) in patients with unstable angina, and 12% (6/49) in controls (all p = NS). CONCLUSIONS: Only a small percentage of patients with CAD demonstrate seropositivity against Chlamydia pneumoniae. Antibiotic therapy in these selected patients, but not in the remaining patients, may be considered rational. These considerations may underlie the failure to see consistent benefits of antibiotic therapy in patients with CAD. PMID- 10803440 TI - Should interventional cardiac catheterization procedures take place at the time of diagnostic procedures? AB - BACKGROUND: In many cardiac catheterization laboratories interventional procedures are performed at a date later than the diagnostic study, causing increased hospital days and costs. Few data exist which compare procedural success, complications, and costs between procedures performed at the time of diagnostic study and those performed later. HYPOTHESIS: The purpose of this study was to evaluate the safety and success of same-day interventional procedures and to quantitate hospital cost savings with this strategy. METHOD: In all, 357 consecutive patients who underwent an elective interventional procedure of a native coronary artery either at the time of diagnostic study (same day, n = 244) or later (delayed, n = 113) were reviewed. Procedural success [< 30% residual lesion post-percutaneous transluminal coronary angioplasty (PTCA) or 0% residual lesion post-stent], major complications [death, emergent coronary artery bypass grafting (CABG), myocardial infarction, and ventricular fibrillation], hospital days, and costs were analyzed. Procedural expense, including the diagnostic and interventional procedure in the cardiac catheterization laboratory, and hospital expense were analyzed. RESULTS: Both groups were similar in terms of age, gender, coronary risk factors, indications (myocardial infarction, unstable angina, abnormal stress test), the culprit coronary artery, type of intervention (PTCA, stent), and lesion complexity (type A, B, C). The average hospital stay for the two groups was 4.37 +/- 2 and 6.55 +/- 2.4 days, respectively (p < 0.0001). The procedural charges were $8,207.99 and 10,581.87, respectively (p < 0.0001). CONCLUSION: Catheter intervention performed at the same time as the diagnostic cardiac catheterization procedure is as successful and as safe as that performed at a later date. Hospital stay and costs, as well as procedural expenses are significantly reduced by this practice. PMID- 10803441 TI - Angiotensin-converting enzyme and apolipoproteins genes polymorphism in coronary artery disease. AB - BACKGROUND: Association between angiotensin-converting enzyme (ACE) as well as apolipoprotein (apo) AI, B, and E polymorphisms and dyslipidemia and coronary artery disease (CAD) is controversial. HYPOTHESIS: This study assessed the distribution of ACE insertion/deletion, apo AI A/G mutation, apo B signal peptide insertion/deletion, apo B XbaI restriction fragment length, and apo E polymorphisms in 388 nondiabetic patients. METHODS: The study population included 112 patients with stable CAD, 139 patients with acute myocardial infarction (AMI), and 137 age-matched control subjects. RESULTS: Univariate analysis showed higher prevalence of XbaI X+/X+ genotype in patients with CAD (p = 0.02). Angiotensin-converting enzyme and apo polymorphisms were not associated with lipid levels or severity of CAD. When all genotypes known to be related to CAD; such as ACE DD, apo AI GG, apo B del/del, and XbaI X+X+, and E4 allele of apo E, were pooled, again no significant differences among groups were seen. Multivariate regression analysis disclosed traditional risk factors and elevated levels of apo B for men and reduced levels of apo AI for women as independent variables for CAD. CONCLUSIONS: In addition to traditional coronary risk factors, apo B and AI could be considered predictors of CAD. No association between either form of CAD and polymorphisms was noted. PMID- 10803442 TI - The use of angiotensin-converting enzyme inhibitors in patients with acute myocardial infarction in community hospitals. Michigan State University Inter Institutional Collaborative Heart (MICH) Study Group. AB - BACKGROUND: Previous studies documenting underutilization of angiotensin converting enzyme inhibitors (ACEIs) in acute myocardial infarction (AMI) have been limited to Medicare populations. HYPOTHESIS: This study examines ACEI prescription rates and predictors in a community sample of hospitalized patients with AMI. METHODS: The charts of 1163 community patients with AMI, prospectively identified at admission between January 1, 1994, and April 30, 1995, were reviewed. RESULTS: Only 64 of 158 (40%) patients considered ideal candidates for ACEI prescription were discharged with a prescription for an ACEI. In a multivariate logistic regression model, prior ACEI utilization [adjusted odds ration (OR) = 3.26; 95% confidence interval (CI) = 2.05-5.20], presence of congestive heart failure (OR = 2.33; CI = 1.50-3.61) and black race (OR = 2.20; CI = 1.34-3.64) were identified as positive predictors of ACEI prescription. Conversely, lack of left ventricular ejection fraction (LVEF) measurement (OR = 0.46; CI = 0.28-0.75), LVEF > 40 ( OR = 0.27; CI = 0.18-0.40), and acute renal failure (OR = 0.08; CI = 0.01-0.44) were negative predictors. Women were also less likely to be discharged with an ACEI prescription (OR = 0.71; CI = 0.48 1.05). Furthermore, women were significantly less likely to have LVEF measured prior to discharge than were males (77 vs. 85%, p = 0.001). CONCLUSION: This study underscores the need for improvement in the utilization of ACEI in eligible patients with AMI. It also identifies opportunities for improvement in prescription rates, especially in women. PMID- 10803443 TI - Correlation of coronary angiography with "tombstoning" electrocardiographic pattern in patients after acute myocardial infarction. AB - BACKGROUND: It has been suggested that a specific pattern of electrocardiographic (ECG) changes following acute myocardial infarction (AMI), the so-called "tombstoning," predicts a poorer outcome, although the cause and associations of such changes are not known. To address the possible cause and implications, we correlated angiographic findings with tombstoning in patients following AMI. METHODS: The study investigated 124 patients with AMI, whose ECGs were taken within 24 h of onset of symptoms and who subsequently underwent angiography. In this population, 24 (19%) patients had a definite tombstoning pattern on their admission ECG. RESULTS: Compared with non-tombstoning ECGs, the significant differences in the tombstoning group are as follows: (1) All patients, including those with inferior infarction, had either total or partial occlusion of the left anterior descending (LAD) artery (100 vs. 44%, p < 0.0001); (2) LAD occlusions were significantly more severe and mostly proximal (100% occlusion: 50 vs. 20.5%, p = 0.02; <50% occlusion: 0 vs. 15.9% p = 0.039; proximal occlusion: 92 vs. 65%, p = 0.017); (3) patients with tombstoning ECGs had a significantly greater incidence of occlusion of all three coronary arteries (54.1 vs. 22%, p = 0.001); (4) tombstoning ECGs were more strongly associated with anterior than with inferior infarction (83.3 vs. 33%, p < 0.0001). CONCLUSIONS: The patients with a tombstoning pattern on the admission ECG, who underwent angiography, were associated with occlusion of a high-grade stenosis of the proximal LAD artery (usually with involvement of more than one artery) and were predominantly seen in association with anterior infarction. PMID- 10803444 TI - Impact of left ventricular size on pharmacologic reverse remodeling in heart failure. AB - BACKGROUND AND HYPOTHESIS: Although medical therapy may normalize echocardiographic left ventricular (LV) systolic function in selected patients with cardiomyopathy, other patients experience no change or a further deterioration in heart failure remodeling. Our aim was to determine what clinical or echocardiographic parameters predict a beneficial therapeutic response. METHODS: We prospectively followed biannual clinical and echocardiographic assessments in 215 patients. Forty-six of these patients ("Nonresponders") experienced no change or a decline in LV ejection fraction at 6 months. Of the 148 patients who improved LV function, 21 ("Responders") normalized LV systolic function at 6 months. Only Responders (n = 21) and Nonresponders (n = 46) were compared. RESULTS: On average, these 67 patients were 54 +/- 12 years old with 4.5 +/- 3.3 years of heart failure. At 6 months, following up-titration of angiotensin-converting enzyme inhibitors and nitrates, Responder LV ejection fraction rose from 22 +/- 6 to 50 +/- 5% with improvement in New York Heart Association classification (2.6 +/- 0.8 to 1.5 +/- 0.8, p = 0.001). These patients had significantly more favorable clinical and echocardiographic outcomes versus Nonresponders despite comparable medical therapy. All baseline demographic, clinical, and echocardiographic variables were equivalent, except for initial LV end-diastolic diameter which differentiated Nonresponders (7.1 +/- 0.7 cm) from Responders (6.1 +/- 0.8 cm), p = 0.007. CONCLUSION: Thus, although heart failure therapy improves LV systolic function in a majority of patients, with normalization in up to 10% of patients, significant LV enlargement may render remodeling unresponsive to pharmacologic intervention, with a potential future need for alternative mechanical or surgical intervention. PMID- 10803445 TI - Exercise-induced QTc-interval changes for predicting improvement in regional blood flow in ischemic myocardium and cardiac output after coronary angioplasty in patients with right bundle-branch block. AB - BACKGROUND: We have previously shown that QT-interval changes are more useful than ST-T changes in evaluating the severity of exercise-induced myocardial ischemia in patients with right bundle-branch block (RBBB). HYPOTHESIS: The purpose of this study was to evaluate whether the improvement in regional myocardial blood flow (RMBF) in ischemic areas and cardiac output after percutaneous transluminal coronary angioplasty (PTCA) can be predicted by exercise-induced QT-interval changes prior to PTCA. METHODS: The RMBF and cardiac output were quantified with nitrogen-13 ammonia positron emission tomography at rest and during exercise in 20 patients with RBBB and ischemic heart disease before and 6 months after PTCA, and in 9 healthy volunteers. RESULTS: Before PTCA, exercise-induced prolongation by < 20 ms or shortening of the Bazett corrected QT (QTc) interval (454 +/- 38 to 451 +/- 41 ms, p = NS) was observed in 13 patients (Group 1) and prolongation by > or = 20 ms (429 +/- 44 to 466 +/- 50 ms, p < 0.002) was observed in 7 (Group 2). The number of regions of exercise induced ischemia was significantly greater in Group 2 than in Group 1 (4.0 +/- 1.2 vs. 2.1 +/- 1.2, p < 0.01). The RMBF in regions of exercise-induced ischemia and cardiac output at rest was not significantly different between Groups 1 and 2, whereas during exercise both the parameters were significantly lower in Group 2 than in Group 1 (both p < 0.05). After successful PTCA, RMBF both at rest and during exercise improved significantly in Group 1 (0.67 +/- 0.04 to 0.71 +/- 0.06 ml/min/g, 0.74 +/- 0.05 to 0.84 +/- 0.08 ml/min/g; both p < 0.0001), but did not improve significantly in Group 2 (0.63 +/- 0.05 to 0.65 +/- 0.07 ml/min/g, 0.65 +/- 0.04 to 0.69 +/- 0.11 ml/ min/g; both p = NS). Cardiac output during exercise improved significantly in Group 1 (6.4 +/- 0.7 to 7.4 +/- 0.9 l/min; p < 0.002) but not in Group 2 (5.7 +/- 0.6 to 5.9 +/- 0.6 l/min; p = NS). CONCLUSIONS: Our results suggest that the marked prolongation of the QTc interval induced by pre PTCA exercise may predict a lack of improvement in RMBF in ischemic areas and cardiac output after PTCA in patients with RBBB and ischemic heart disease. PMID- 10803446 TI - Cardiac sympathetic activity in the asymmetrically hypertrophied septum in patients with hypertension or hypertrophic cardiomyopathy. AB - BACKGROUND: In patients with essential hypertension (HT), proportional (symmetric) left ventricular hypertrophy (LVH) is common. In contrast, hypertrophic cardiomyopathy (HCM) is characterized by disproportional LVH and, in particular, asymmetric septal hypertrophy (ASH); however, some hypertensive patients also develop ASH. It has not been determined whether such cases represent a distinct type of hypertensive LVH or HCM combined with hypertension. HYPOTHESIS: The study was undertaken to evaluate sympathetic activity in the interventricular septum in patients with HT and ASH or in patients with HCM. METHODS: The patients were evaluated by I-123 meta-iodobenzylguanidine (MIBG) and thallium-201 (201Tl) single-photon emission computed tomography (SPECT), respectively. They were divided into three groups: patients with essential HT and symmetric septal hypertrophy (Group A), patients with HT and ASH (Group B), and patients with HCM and ASH (Group C). RESULTS: Compared with the lateral wall, early uptake of MIBG in the septum was significantly higher in Group B than in Group A, but not significantly different between Groups A and C. Compared with the lateral wall, early uptake of 201Tl in the septum did not differ among the three groups. No significant difference in the MIBG clearance in the lateral wall was seen among the three groups. By contrast, MIBG clearances in the septum and apex were significantly greater in Group C than in Groups A and B. There was an inverse correlation between systolic thickening and MIBG clearance in the septum. CONCLUSION: These findings suggest that sympathetic activity in the septum differs between patients with HT and ASH and patients with HCM. PMID- 10803447 TI - Influence of autonomic neuropathy upon left ventricular dysfunction in insulin dependent diabetic patients. AB - BACKGROUND: Diabetic cardiomyopathy is a well-defined complication of diabetes that occurs in the absence of ischemic, vascular, and hypertensive disease. HYPOTHESIS: The study was undertaken to test the relationship among autonomic neuropathy (AN), 24-h blood pressure (BP) profile, and left ventricular function. METHODS: Nineteen type-1 diabetic patients underwent autonomic tests and echocardiographic examination. Patients were divided according to the presence (AN+) or absence (AN-) of AN. RESULTS: In the AN+ group (n = 8), the E/A ratio at echo was lower than in the AN- group (n = 11) (1.1 +/- 0.3 vs. 1.6 +/- 0.3; p < 0.005). Systolic and diastolic BP reductions during sleep were smaller in the AN+ than in the AN- group (6.6 +/- 6.6 vs. 13.0 +/- 4.3%; p < 0.03 for systolic and 12.8 +/- 6.8 vs. 20.0 +/- 4.0% for diastolic BP reduction; p < 0.03, respectively). Considering all patients, the E/A ratio correlated inversely with awake diastolic BP (r - 0.63; p = 0.005); sleep systolic BP (r - 0.48; p = 0.04), and sleep diastolic BP (r - 0.67; p = 0.002). The AN correlated with diastolic interventricular septum thickness (r 0.57; p = 0.01), sleep systolic BP (r 0.45; p = 0.05), sleep diastolic BP (r 0.54; p = 0.02), and correlated inversely with systolic and diastolic sleep BP reduction (r - 0.49; p = 0.03 and r - 0.67; p = 0.002, respectively). Finally, E/A ratio and AN score correlated between themselves (r - 0.6; p = 0.005). CONCLUSION: Our results suggest that left ventricular diastolic dysfunction may be detected very early in type-1 diabetic patients with AN. Parasympathetic lesion and nocturnal elevations in BP could be the link between AN and diastolic ventricular dysfunction. PMID- 10803448 TI - Acute myocardial infarction--a late complication of intracoronary stent placement. AB - BACKGROUND: Myocardial infarction (MI) as the first indication of postangioplasty restenosis is extremely rare, and it has been speculated that the fibroproliferative restenotic lesion is less likely to undergo plaque rupture than the lipid-laden native atherosclerotic lesion. HYPOTHESIS: The present study was designed to examine whether intracoronary stent implantation affects this course. METHODS: In all, 994 consecutive patients who underwent angioplasty and intracoronary stent implantation in our hospital were reviewed retrospectively for the occurrence of MI. RESULTS: Eight patients (0.8%), all male and hypertensive, aged 33-83 years, presented with an MI due to stent occlusion more than 30 days following stenting (range: 35-398 days). In two patients, MI occurred 3 and 5 h, respectively, following completion of a maximal high-level exercise test that was negative for ischemia. Angiography revealed complete occlusion or significant stenosis of the stent in all eight patients, with an obvious intimal dissection in either edge of the stent in six patients. Except for gender and hypertension, no correlation was found with other risk factors, vessel involved, initial angiographic results, or with stent design, diameter, or length. CONCLUSIONS: Myocardial infarction as a late complication of successful stent implantation occurred in 0.8% of our patients. This is only the lower bound of the estimated frequency for such an event. We hypothesize that the transition point between the relatively fixed stent and the normal artery is exposed to high deformation stress which makes it vulnerable to rupture and dissection. Strenuous exercise and hypertension may increase the deformation stress and the risk of intimal rupture. PMID- 10803449 TI - Ethnicity does not affect outcomes of coronary angioplasty. AB - BACKGROUND: Access to high quality medical care and especially to complex procedures may be adversely affected in members of a minority ethnic group or a lower socioeconomic class. For example, Caucasians undergo coronary artery bypass grafting (CABG) or percutaneous transluminal coronary interventions (PTCI) twice as frequently as African-Americans. Data exist to suggest that African-Americans derive less benefit than Caucasians from CABG. HYPOTHESIS: We investigated the possibility that outcomes of catheter-based coronary angioplasty might also be less favorable in minority populations. METHODS: We analyzed in-hospital outcomes in 6,559 consecutive patients who underwent PTCI in our laboratory. In 37 ethnicity was classified as "other," 5,203 (79.8%) were identified as Caucasians, 863 (13.2%), as African-Americans, and 456 (7.0%), as Hispanics. Twelve baseline clinical, angiographic, and procedural characteristics were entered into a computerized data base. Hospital complications were identified by trained quality assurance nurses. RESULTS: Substantial differences in baseline characteristics existed between the populations. Despite these differences, on univariate comparison of ethnicity and outcome, no differences between ethnic groups were found with a single exception. Mortality in Hispanics was higher than in the other two populations. (2.0 vs. 0.7 and 0.8%, respectively, p = 0.008). However, when this was adjusted for baseline characteristics, the difference was not significant. CONCLUSIONS: In contrast to previous studies suggesting less favorable outcomes of CABG in African-American patients, this analysis demonstrates an equal frequency of procedural success and rate of hospital complications for PTCI in that population, in Hispanics, and in Caucasians. PMID- 10803450 TI - Images in cardiology. Cardiac involvement in lightning strike injury. PMID- 10803451 TI - Fungal endocarditis at the aortotomy site after aortic valve replacement. AB - This brief report presents a patient with fungal endocarditis involving the suture location in the ascending aorta after aortotomy to replace a stenotic aortic valve. It emphasizes the importance of expanding the diagnostic investigation for endocarditis beyond the prosthesis, which was normal in this case. It also reiterates the value of transesophageal echocardiography during the evaluation of prosthetic valves and reminds the echocardiographer to visualize the aortotomy site carefully. PMID- 10803452 TI - Adolph Fick: mathematician, physicist, physiologist. PMID- 10803453 TI - Metabolism and pharmacokinetics of oxazaphosphorines. AB - The 2 most commonly used oxazaphosphorines are cyclophosphamide and ifosfamide, although other bifunctional mustard analogues continue to be investigated. The pharmacology of these agents is determined by their metabolism, since the parent drug is relatively inactive. For cyclophosphamide, elimination of the parent compound is by activation to the 4-hydroxy metabolite, although other minor pathways of inactivation also play a role. Ifosfamide is inactivated to a greater degree by dechloroethylation reactions. More robust assay methods for the 4 hydroxy metabolites may reveal more about the clinical pharmacology of these drugs, but at present the best pharmacodynamic data indicate an inverse relationship between plasma concentration of parent drug and either toxicity or antitumour effect. The metabolism of cyclophosphamide is of particular relevance in the application of high dose chemotherapy. The activation pathway of metabolism is saturable, such that at higher doses (greater than 2 to 4 g/m2) a greater proportion of the drug is eliminated as inactive metabolites. However, both cyclophosphamide and ifosfamide also act to induce their own metabolism. Since most high dose regimens require a continuous infusion or divided doses over several days, saturation of metabolism may be compensated for, in part, by auto induction. Although a quantitative distinction may be made between the cytochrome P450 isoforms responsible for the activating 4-hydroxylation reaction and those which mediate the dechloroethylation reactions, selective induction of the activation pathway, or inhibition of the inactivating pathway, has not been demonstrated clinically. Mathematical models to describe and predict the relative contributions of saturation and autoinduction to the net activation of cyclophosphamide have been developed. However, these require careful validation and may not be applicable outside the exact regimen in which they were derived. A further complication is the chiral nature of these 2 drugs, with some suggestion that one enantiomer may have a favourable profile of metabolism over the other. That the oxazaphosphorines continue to be the subject of intensive investigation over 30 years after their introduction into clinical practice is partly because of their antitumour activity. Further advances in analytical and molecular pharmacological techniques may further optimise their use and allow rational design of more selective analogues. PMID- 10803454 TI - Magnesium sulfate in eclampsia and pre-eclampsia: pharmacokinetic principles. AB - Magnesium sulfate (MgSO4) is the agent most commonly used for treatment of eclampsia and prophylaxis of eclampsia in patients with severe pre-eclampsia. It is usually given by either the intramuscular or intravenous routes. The intramuscular regimen is most commonly a 4 g intravenous loading dose, immediately followed by 10 g intramuscularly and then by 5 g intramuscularly every 4 hours in alternating buttocks. The intravenous regimen is given as a 4 g dose, followed by a maintenance infusion of 1 to 2 g/h by controlled infusion pump. After administration, about 40% of plasma magnesium is protein bound. The unbound magnesium ion diffuses into the extravascular-extracellular space, into bone, and across the placenta and fetal membranes and into the fetus and amniotic fluid. In pregnant women, apparent volumes of distribution usually reach constant values between the third and fourth hours after administration, and range from 0.250 to 0.442 L/kg. Magnesium is almost exclusively excreted in the urine, with 90% of the dose excreted during the first 24 hours after an intravenous infusion of MgSO4. The pharmacokinetic profile of MgSO4 after intravenous administration can be described by a 2-compartment model with a rapid distribution (a) phase, followed by a relative slow beta phase of elimination. The clinical effect and toxicity of MgSO4 can be linked to its concentration in plasma. A concentration of 1.8 to 3.0 mmol/L has been suggested for treatment of eclamptic convulsions. The actual magnesium dose and concentration needed for prophylaxis has never been estimated. Maternal toxicity is rare when MgSO4 is carefully administered and monitored. The first warning of impending toxicity in the mother is loss of the patellar reflex at plasma concentrations between 3.5 and 5 mmol/L. Respiratory paralysis occurs at 5 to 6.5 mmol/L. Cardiac conduction is altered at greater than 7.5 mmol/L, and cardiac arrest can be expected when concentrations of magnesium exceed 12.5 mmol/L. Careful attention to the monitoring guidelines can prevent toxicity. Deep tendon reflexes, respiratory rate, urine output and serum concentrations are the most commonly followed variables. In this review, we will outline the currently available knowledge of the pharmacokinetics of MgSO4 and its clinical usage for women with pre-eclampsia and eclampsia. PMID- 10803456 TI - Pharmacokinetic drug interactions between oral contraceptives and second generation anticonvulsants. AB - Drug interactions between oral contraceptives (OCs) and traditional anticonvulsants have been well described. However, in the past decade, a number of new anticonvulsants have been developed, as well as modifications made in the composition of the OC preparations themselves. Additionally, anticonvulsants are increasingly employed in the therapy of nonseizure-related disorders, placing more women at risk of potential drug interactions that may lead to contraceptive failure. Second-generation anticonvulsants include felbamate, gabapentin, lamotrigine, oxcarbazepine, tiagabine, topiramate, vigabatrin and zonisamide. Most have been approved for adjunctive management of seizures refractory to therapy with traditional anticonvulsants. On the basis of available study data in women receiving concomitant OC preparations, gabapentin, lamotrigine, tiagabine and vigabatrin may be administered without significant pharmacokinetic interactions that potentially diminish contraceptive efficacy. However, additional or alternative contraceptive measures, including using OCs with higher estrogen content, are recommended when using felbamate, oxcarbazepine and topiramate, as these agents have demonstrated enzyme-inducing activity leading to reduced plasma steroid concentrations. The effects of zonisamide in women receiving OCs have yet to be reported. It is important to characterise the properties [e.g. substrate and enzyme activity (particularly cytochrome P450 3A4 induction)] of new anticonvulsants and recognise their potential to interfere with OCs. However, a pharmacokinetic interaction does not in itself indicate loss of OC efficacy. Contraceptive failure should be measured by changes in ovarian hormone concentrations, maturation of ovarian follicle(s) or ovulation. PMID- 10803455 TI - Adaptive control methods for the dose individualisation of anticancer agents. AB - Numerous studies have found a clear relationship between systemic exposure and the toxicity or (more rarely) the efficacy of anticancer agents. Moreover, the clearance of most of these drugs differs widely between patients. These findings, combined with the narrow therapeutic index of anticancer drugs, suggest that patient outcome would be improved if doses were individualised to achieve a target systemic exposure. Bayesian maximum a posteriori probability (MAP) forecasting is an efficient and robust method for the optimisation of drug therapy, but its use for anticancer drugs is not yet extensive. The aim of this paper is to review the application of population pharmacokinetics and MAP to anticancer drugs and to evaluate whether and when MAP Bayesian estimation improves the clinical benefit of anticancer chemotherapy. For each drug, the relationships between pharmacokinetic variables [e.g. plasma concentration or the area under the concentration-time curve] and pharmacodynamic effects are described. Secondly, the methodologies employed are considered and, finally, the results are analysed in terms of predictive performance as well as, where possible, the impact on clinical end-points. Some studies were retrospective and intended only to evaluate individual pharmacokinetic parameter values using very few blood samples. Among the prospective trials, a few studied the pharmacokinetic/pharmacodynamic relationships which provided the basis for routine pharmacokinetic monitoring. Others were performed in clinical context where MAP Bayesian estimation was used to determine maximum tolerated systemic exposure (e.g. for carboplatin, topotecan, teniposide) or for pharmacokinetic monitoring (e.g. for methotrexate or platinum compounds). Indeed, its flexibility in blood sampling times makes this technique much more applicable than other limited sampling strategies. These examples demonstrate that individual dose adjustment helps manage toxicity. The performance of pharmacokinetic monitoring is linked to the methodology used at each step of its design and application. Moreover, a limitation to the use of pharmacokinetic monitoring for certain anticancer drugs has been the difficulty in obtaining pharmacokinetic or pharmacodynamic data. Recent progress in analytical methods, as well as the development of noninvasive methods (such as positron emission tomography) for evaluating the effects of chemotherapy, will help to define pharmacokinetic pharmacodynamic relationships. Bayesian estimation is the strategy of choice for performing pharmacokinetic studies, as well as ensuring that a given patient benefits from the desired systemic exposure. Together, these methods could contribute to improving cancer chemotherapy in terms of patient outcome and survival. PMID- 10803457 TI - A concept for pharmacokinetic-pharmacodynamic dosage adjustment in renal impairment: the case of aminoglycosides. AB - BACKGROUND: For patients with impaired renal function, dosage adjustment is necessary for many drugs. Adjustment with respect only to pharmacokinetic parameters may be insufficient. OBJECTIVE: To apply the theory of pharmacokinetics and pharmacodynamics to derive a mathematical model that links the concentration-time course and the clinical response by means of the pharmacokinetic-pharmacodynamic parameter 'area under the effect-time curve' (AUETC), and to use this analysis and clinical data for aminoglycosides to calculate dosage adjustments in renal impairment. METHODS: Model parameters were estimated for the antimicrobial and nephrotoxic effects of aminoglycosides on the basis of data from the literature. Effect parameters were calculated for various degrees of impaired renal function. RESULTS: Use of the model parameters gave a high correlation between the predicted and the observed (literature) values for antimicrobial efficacy and nephrotoxicity. When calculating dosage adjustments in renal impairment, it was possible to hold only one effect (antimicrobial or nephrotoxic) constant by dosage adjustment, whereas the other changed unfavourably. This was explained by differences between the pharmacodynamic parameters for each effect. For high antimicrobial efficacy, a target peak concentration of 9 mg/L (for gentamicin) should be obtained every 48 hours in advanced renal impairment. For low nephrotoxicity, the peak concentration should not exceed 3 mg/L. CONCLUSIONS: The parameter AUETC could be a useful pharmacokinetic-pharmacodynamic surrogate marker for dosage adjustment in renal impairment. Using the AUETC method, the beneficial effect can be balanced against the adverse effect. PMID- 10803458 TI - Hsp27 functions as a negative regulator of cytochrome c-dependent activation of procaspase-3. AB - The release of mitochondrial cytochrome c by genotoxic stress induces the formation of a cytosolic complex with Apaf-1 (mammalian CED4 homolog) and thereby the activation of procaspase-3 (cas-3) and procaspase-9 (cas-9). Here we demonstrate that heat-shock protein 27 (Hsp27) inhibits cytochrome c (cyt c) dependent activation of cas-3. Hsp27 had no effect on cyt c release, Apaf-1 and cas-9 activation. By contrast, our results show that Hsp27 associates with cas-3, but not Apaf-1 or cas-9, and inhibits activation of cas-3 by cas-9-mediated proteolysis. Furthermore, the present results demonstrate that immunodepletion of Hsp27 depletes cas-3. Importantly, treatment of cells with DNA damaging agents dissociates the Hsp27/cas-3 complex and relieves inhibition of cas-3 activation. These findings define a novel function for Hsp27 and provide the first evidence that a heat shock protein represses cas-3 activation. PMID- 10803459 TI - Retinoblastoma susceptibility protein, Rb, possesses multiple BRCT-Ws, BRCA1 carboxyl-terminus-related W regions with DNA break-binding activity. AB - The BRCT region, the carboxyl-terminus of BRCA1 (the breast cancer susceptibility gene 1 product), is ubiquitous in several proteins that participate in cell cycle checkpoints and DNA repair. We have previously shown that the BRCT regions of TopBP1 (DNA topoisomerase II binding protein 1) and BRCA1 bound DNA breaks. A BRCT-related region, BRCT-W1, in the retinoblastoma susceptibility gene product (Rb) also could bind DNA fragments, independently of DNA sequences. Five BRCT-W regions were found in the Rb family. All BRCT-Ws of Rb bound DNA fragments. Electron microscopy and treatment with an exonuclease showed that BRCT-Ws bound double-strand DNA breaks. Since some BRCTs are exceptional common relating elements in tumor suppression, our findings reveal novel aspects of the tumor suppression mechanism. PMID- 10803460 TI - Differential interaction of plakoglobin and beta-catenin with the ubiquitin proteasome system. AB - Beta-catenin and plakoglobin are closely related armadillo family proteins with shared and distinct properties; Both are associated with cadherins in actin containing adherens junctions. Plakoglobin is also found in desmosomes where it anchors intermediate filaments to the desmosomal plaques. Beta-catenin, on the other hand, is a component of the Wnt signaling pathway, which is involved in embryonic morphogenesis and tumorigenesis. A key step in the regulation of this pathway involves modulation of beta-catenin stability. A multiprotein complex, regulated by Wnt, directs the phosphorylation of beta-catenin and its degradation by the ubiquitin-proteasome system. Plakoglobin can also associate with members of this complex, but inhibition of proteasomal degradation has little effect on its levels while dramatically increasing the levels of beta-catenin. Beta-TrCP, an F-box protein of the SCF E3 ubiquitin ligase complex, was recently shown to play a role in the turnover of beta-catenin. To elucidate the basis for the apparent differences in the turnover of beta-catenin and plakoglobin we compared the handling of these two proteins by the ubiquitin-proteasome system. We show here that a deletion mutant of beta-TrCP, lacking the F-box, can stabilize the endogenous beta-catenin leading to its nuclear translocation and induction of beta-catenin/LEF-1-directed transcription, without affecting the levels of plakoglobin. However, when plakoglobin was overexpressed, it readily associated with beta-TrCP, efficiently competed with beta-catenin for binding to beta-TrCP and became polyubiquitinated. Fractionation studies revealed that about 85% of plakoglobin in 293 cells, is Triton X-100-insoluble compared to 50% of beta catenin. These results suggest that while both plakoglobin and beta-catenin can comparably interact with beta-TrCP and the ubiquitination system, the sequestration of plakoglobin by the membrane-cytoskeleton system renders it inaccessible to the proteolytic machinery and stabilizes it. PMID- 10803461 TI - LMP1 of Epstein-Barr virus suppresses cellular senescence associated with the inhibition of p16INK4a expression. AB - Epstein-Barr virus is associated with a number of human proliferative and malignant diseases. It is capable of immortalizing human primary B-lymphocytes in vitro. Studies indicate that latent membrane protein LMP1 is one of the viral proteins essential for this process. In this report, LMP1 was shown to prevent primary mouse embryonic fibroblasts from entering into replicative senescence in vitro. It further suppresses the senescence-associated induction of p16INK4a, commonly believed to be a key regulator of replicative senescence. In addition, LMP1 was shown to prevent premature senescence provoked by oncogenic ras in mouse embryonic fibroblasts, and to inhibit the oncogene ras-mediated induction of p16INK4a and p21WAF1. In parallel, LMP1 also prevents ras-induced premature senescence in rat embryonic fibroblasts REF52 and human diploid fibroblasts IMR90. Moreover, LMP1 is capable of suppressing the p16INK4a promoter in REF52 and Saos-2 cells in a promoter reporter assay. Our findings suggest that with the expression of p16INK4a and replicative senescence being suppressed, LMP1 may play a key role in Epstein-Barr virus-associated proliferative diseases, and it may further contribute to cancer development by preventing premature senescence induced by mitogenic oncogenes. PMID- 10803462 TI - R-Ras3, a brain-specific Ras-related protein, activates Akt and promotes cell survival in PC12 cells. AB - The GTP-binding protein, R-Ras3/M-Ras, is a novel member of the Ras subfamily of GTPases which shows highest sequence similarity to the TC21 gene. R-Ras3 is highly expressed in both human and mouse brain and ectopic expression of a constitutively active mutant of R-Ras3 induces cellular transformation in NIH3T3 cells. To gain further insight into the normal cellular function of R-Ras3, we examined the ability of R-Ras3 in activating several known intracellular signaling cascades. We observed that R-Ras3 is a relatively weak activator of the mitogen-activated protein kinase/extracellular-signal-regulated kinases (MAPK/ERKs) when compared to the H-Ras oncogene. On the contrary, both R-Ras3 and H-Ras activated the Jun N-terminal kinase (JNK) to a similar extent. Under similar experimental conditions, R-Ras3 significantly stimulated one of the phosphatidylinositol 3-kinase (PI3-K) downstream substrates, Akt/PKB/RAC (Akt), which has been extensively implicated in mediating cell survival signaling. The activation of Akt by R-Ras3 was most likely to be PI3-K-dependent since this biochemical event was blocked by the pharmacological inhibitors, Wortmannin and LY294002, as well as by a dominant negative mutant of PI3-K. More importantly, R Ras3 affinity-precipitated PI3-K from cell extracts in a GTP-dependent manner, and associated lipid kinase activity was readily detectable in R-Ras3 immune complexes. The biological significance of R-Ras3 in inducing Akt kinase activity is evidenced by the ability of an activated R-Ras3 to confer cell survival in the rat pheochromocytoma cell line, PC12. As expected, this biological activity of R Ras3 was also abrogated by the addition of LY294002. Thus, R-Ras3 represents a novel G-protein which may play a role in cell survival of neural-derived cells. PMID- 10803463 TI - Nonrandom 4p13 rearrangements of the RhoH/TTF gene, encoding a GTP-binding protein, in non-Hodgkin's lymphoma and multiple myeloma. AB - We recently isolated the RhoH/TTF gene by its fusion to the LAZ3/BCL6 gene, in a non-Hodgkin's lymphoma (NHL) cell line, which bore a t(3;4)(q27;p11-13) translocation. This gene encodes a novel Rho GTP-binding protein and is specifically expressed in hematopoietic tissues. We made its precise mapping at band 4p13, and described its partial genomic structure. Using fluorescence in situ hybridization and molecular analyses, we report here on the rearrangement of the RhoH/TTF gene, at band 4p13, in four cases of NHL with t(3;4)(q27;p13) translocation and its fusion to the LAZ3/BCL6 gene at band 3q27, in three of these cases. RT-PCR analysis of two cases allowed the detection of variable fusion transcripts emerging from the rearranged alleles, and in one case, a deregulated expression of both RhoH/TTF and LAZ3/BCL6 genes, by promoter substitution, was observed. We also show here another rearrangement of the RhoH/TTF gene in a patient with multiple myeloma and t(4;14)(p13;q32) translocation, with breakage within the IGH gene. It is the first report which describes the recurrent chromosomal alteration of a GTP-binding protein encoding gene, in patients with hematopoietic malignancies. PMID- 10803464 TI - A new feature of Mpl receptor: ligand-induced transforming activity in FRE rat fibroblasts. AB - Mpl is the receptor for thrombopoietin, the primary regulator of platelet production by megakaryocytes. Upon stimulation by its ligand, Mpl receptor induces proliferation and differentiation of hematopoietic cell lines of various origins. In this paper, we show that Mpl is also able to transform FRE rat fibroblasts in the presence of MGDF (pegylated Megakaryocyte Growth and Development Factor), a modified form of its ligand. We also demonstrate that upon MGDF stimulation Mpl receptor activates the classical transduction pathways described for hematopoietic cell lines in FRE cells. Introduction of Mpl deletion mutants in FRE cells allowed us to demonstrate that the C-terminal region of the Mpl intracytoplasmic domain, which is involved in hematopoietic differentiation, is necessary for the transformation process. Within that region, site-directed mutagenesis showed that the Y112 residue, which is required for Shc phosphorylation, is essential for rat fibroblast transformation by Mpl/MGDF, suggesting the involvement of Shc in Mpl-mediated transformation. Interestingly, we showed that transformation correlated with strong and sustained MAPK activation. Neither Jak2, Stat3 nor Stat5 phosphorylation was sufficient to induce the transformation process. Taken altogether, our results suggest the oncogenicity of Mpl in fibroblastic cells in the presence of its ligand. PMID- 10803465 TI - Molecular dissection of TrkA signal transduction pathways mediating differentiation in human neuroblastoma cells. AB - Activation of the neurotrophin receptor TrkA by its ligand nerve growth factor (NGF) initiates a cascade of signaling events leading to neuronal differentiation in vitro and might play an important role in the differentiation of favorable neuroblastomas (NB) in vivo. To study TrkA signal transduction pathways and their effects on differentiation in NB, we stably expressed wild-type TrkA and five different TrkA mutants in the NGF unresponsive human NB cell line SH-SY5Y. Resulting clones were characterized by TrkA mRNA and protein expression, and by autophosphorylation of the receptor. Introduction of wild-type TrkA restored NGF responsiveness of SH-SY5Y cells, as demonstrated by morphological differentiation, activation of mitogen-activated protein kinases (MAPK) and induction of immediate-early genes. Expression of TrkA in the absence of NGF resulted in growth inhibition of transfectants compared to parental cells, whereas NGF-treatment increased their proliferation rate. Analysis of downstream signal transduction pathways indicated that NGF-induced differentiation was dependent on TrkA kinase activity. Our data suggest that several redundant pathways are present further downstream, but activation of the RAS/MAPK signaling pathway seems to be of major importance for NGF mediated differentiation of NB cells. Our results also show that the signaling effector SH2-B is a substrate of NGF-mediated Trk signaling in NB, whereas it is not activated by NGF in rat pheochromocytoma PC12 cells. This might explain the differences we observed in TrkA signaling between neuroblastoma and PC12 cells. Further insight into TrkA signaling may suggest new options for the treatment of NB. PMID- 10803466 TI - Evidence for an interaction between the insulin receptor and Grb7. A role for two of its binding domains, PIR and SH2. AB - The molecular adapter Grb7 is likely to be implicated in the development of certain cancer types. In this study we show that Grb7 binds the insulin receptors, when they are activated and tyrosine phosphorylated. This interaction is documented by two-hybrid experiments, GST pull-down assays and in vivo coimmunoprecipitations. In addition, our results argue in favor of a preferential association between Grb7 and the insulin receptors when compared to other tyrosine kinase receptors like the EGF receptor, the FGF receptor and Ret. Interestingly, Grb7 is not a substrate of the insulin receptor tyrosine kinase activity. Grb7 binds the activated tyrosine kinase loop of the insulin receptors. Two domains of Grb7 are implicated in the insulin receptor binding: the SH2 domain and the PIR (phosphotyrosine interacting region). The role of these two domains in the interaction with the insulin receptor was already reported for Grb10 and Grb14, the other members of the Grb7 family of proteins. However, the relative importance of these domains varies, considering the receptor and the Grb protein. These differences should be a determinant of the specificity of the receptor tyrosine kinase-Grbs binding, and thus of the implication of Grb7/10/14 in signal transduction. PMID- 10803467 TI - Somatic mitochondrial DNA (mtDNA) mutations in papillary thyroid carcinomas and differential mtDNA sequence variants in cases with thyroid tumours. AB - Somatic mutations in mtDNA have recently been identified in colorectal tumours. Studies of oncocytic tumours have led to hypotheses which propose that defects in oxidative phosphorylation may result in a compensatory increase in mitochondrial replication and/or gene expression. Mutational analysis of mtDNA in thyroid neoplasia, which is characterised by increased numbers of mitochondria and is also one of the most common sites of oncocytic tumours. has been limited to date. Using the recently developed technique of two-dimensional gene scanning, we have successfully examined 21 cases of thyroid tumours, six cases of non-neoplastic thyroid pathology, 30 population controls, nine foetal thyroid tissues and nine foetal tissues of non-thyroid origin, either kidney or liver. We have identified three different somatic mutations (23%) in papillary thyroid carcinomas. In addition, we have found significant differential distributions of mtDNA sequence variants between thyroid carcinomas and controls. Interestingly, these variants appear to be more frequent in the genes which encode complex I of the mitochondrial electron transport chain compared to normal population controls. These findings suggest first, that somatic mtDNA mutations may be involved in thyroid tumorigenesis and second, that the accumulation of certain non-somatic variants may be related to tumour progression in the thyroid. PMID- 10803468 TI - Conserved region 2 of adenovirus E1A has a function distinct from pRb binding required to prevent cell cycle arrest by p16INK4a or p27Kip1. AB - Ectopic expression of the CDK inhibitors (CKIs) p16INK4a and p27Kip1 in Rat1 fibroblasts induces dephosphorylation and activation of Retinoblastoma-family proteins (pRb, p107 and p130), their association with E2F proteins, and cell cycle arrest in G1. The growth-inhibitory action of p16, in particular, is believed to be mediated essentially via pRb activation. The 12S E1A protein of human Adenovirus 5 associates with pRb-family proteins via residues in its Conserved Regions (CR) 1 and 2, in particular through the motif LXCXE in CR2. These interactions are required for E1A to prevent G1 arrest upon co-expression of CKIs. We show here that mutating either of two conserved motifs adjacent to LXCXE in CR2, GFP and SDDEDEE, also impairs the ability of E1A to overcome G1 arrest by p16 or p27. Strikingly, however, these mutations affect neither the association of E1A with pRb, p07 and p130, nor its ability to derepress E2F-1 transcriptional activity in transient transfection assays. One of the EIA mutants, however, is defective in derepressing several endogenous E2F target genes in the presence of p16 or p27. Thus, CR2 possesses an essential function besides pRb-binding. We speculate that this function might be required for the full derepression of E2F-regulated genes in their natural chromatin context. PMID- 10803469 TI - Left ventricular mass in type 2 diabetes mellitus. A study employing a simple ECG index: the Cornell voltage. AB - Both type 2 diabetes mellitus (DM2) and left ventricular hypertrophy are associated with an increased risk of cardiovascular diseases (CVD). A strong association between hyperinsulinemia, which is the hallmark of DM2 and of insulin resistance syndrome (a cohort of metabolic abnormalities such as DM2, dyslipidemia, hyperuricemia, obesity, hypertension, hyperfibrinogenemia), and left ventricular (LV) hypertrophy was found in several studies. We studied 140 consecutive (both normo- and hypertensive) DM2 patients to determine a possible link between metabolic features and the degree of LV mass, calculated by the ECG method of Cornell voltage. The Cornell voltage value was 12.9+/-0.5 mm in the DM2 population as a whole, and 13.6+/-0.7 vs 11.7+/-0.9 mm (p=NS) in hypertensive and normotensive DM2 subgroups, respectively. Among all the metabolic parameters taken into account, the multivariate analysis shows that the fasting plasma insulin level is the strongest independent predictor of LV mass, both in the whole population (p=0.0005) and in the normo (p=0.0460) and hypertensive DM2 (p=0.0184) subgroups. PMID- 10803470 TI - Markers of activation of coagulation and fibrinolysis in patients with Cushing's syndrome. AB - Patients with active Cushing's syndrome have an increased thrombotic tendency. We chose to reassess the mechanism underlying the thrombophilic state associated with this clinical condition using sensitive markers of coagulation and fibrinolysis activation in 17 patients with active disease. The results were compared with those obtained in 12 Cushing's patients successfully treated by surgery and in 20 normal individuals. The general pattern of results in patients with active disease was the finding of increased levels of von Willebrand factor (VWF: Ag), a marker of enhanced metabolic function of endothelial cells (VWF:Ag 181 +/- 42 vs 110 +/- 43, p<0.001 in normal subjects), accompanied by signs of heightened thrombin and plasmin generation, expressed by high levels of thrombin antithrombin (TAT 5.59+/-3.6 vs 3.06+/-0.92 ng/ml in controls, p<0.01) and plasmin-antiplasmin complexes (PAP 407+/-176 vs 245+/-67 ng/ml in controls, p<0.01). VWF:Ag and TAT values were significantly higher in hypertensive than in normotensive patients with active disease (205+/-40 vs 155+/-26 U/dl, p<0.05 and 7.49+/-3.7 vs 3.45+/-1.8, p<0.01, respectively). Plasma levels of plasminogen activator inhibitor type 1 were higher, though not to a statistically significant extent, in patients with active disease compared to controls (12.8+/-12.3 vs 5.6+/-7.4 IU/ml, NS) and positively correlated with body mass index (r=0.66, p<0.01). After surgical control of Cushing's syndrome, there was a partial or complete reversal of the abnormalities to values similar to those found in normal individuals. Our data suggest that the thrombophilic state present in patients with active Cushing's syndrome is related to an enhanced metabolic function of endothelial cells; this in turn may be caused by an heightened production of thrombin with secondary hyperfibrinolysis. Primary prophylaxis with anticoagulants is recommended in these patients when they are exposed to a thrombophilic condition such as surgery. PMID- 10803471 TI - Comparison of the effects of antiprogestins RU38486, ZK98299 and ORG31710 on periovulatory hypophysial, ovarian and adrenal hormone secretion in the rat. AB - The antiprogestin (AP) RU38486 (RU) blocks progesterone (P) and glucocorticoid (G) actions. Administration of 4 mg RU on proestrous morning to cyclic rats dissociates LH and FSH secretion on proestrous afternoon, early estrus and on estrous afternoon. In order to ascertain which action blocked by RU is predominant in the control of periovulatory LH and FSH secretion, a study was made on the effects of: a) 1 or 4 mg of ZK98299 (ZK) (type I P antagonist; Schering), b) 2 or 8 mg of Org31710 (OR) (type II P antagonist lacking anti-G actions; Organon) or c) 1 or 4 mg of RU (type II P antagonist; Exelgyn) to 4-day cyclic rats on proestrous morning on serum concentrations of LH, FSH, inhibin alpha (I), estradiol-17beta (E), progesterone (P) and corticosterone (B) at 18:30 h on proestrus and at 02:00 and 18:30 h on estrus. Controls, receiving 0.2 ml oil, had elevated serum concentrations of all six hormones on proestrous afternoon; at early estrus, only serum concentrations of FSH and P remained elevated, and, on estrous afternoon, all hormones but I and B, that peaked again, had reached their lowest serum levels. All AP treatments except 1 mg ZK had the same effects. On proestrous afternoon serum LH concentrations were reduced and serum FSH concentrations were suppressed whereas serum levels of I, E, P and B were unaffected. At early estrus, basal serum concentrations of LH and E increased while FSH secretion was abolished. Serum levels of I, P and B did not differ from controls. AP treatments increased basal LH concentration, hyperstimulated FSH secretion and reduced serum I concentration on the afternoon of estrus. E, P and B serum levels did not differ from controls at this stage. Treatment with 1 mg ZK was less effective in reducing serum FSH on proestrous afternoon and at early estrus, and had no effect on serum concentrations of any hormone on estrous afternoon. These results indicate that blockade of P receptor activation by P is, predominantly, the mechanism of AP action on periovulatory gonadotropin secretion in rats. PMID- 10803472 TI - The negative GH auto-feedback in childhood: effects of rhGH and/or GHRH on the somatotroph response to GHRH or hexarelin, a peptidyl GH secretagogue, in children. AB - Aim of the present study was to further clarify the negative GH auto-feedback mechanisms in childhood. To this goal we studied the effects of rhGH and/or GHRH administration on the GH response to GHRH or hexarelin (HEX), a peptidyl GH secretagogue, in normal short children. In 34 prepubertal children (12 girls and 22 boys, age 8.2- 14.2 yr) with normal short stature (normal height velocity and IGF-I levels) the following tests were performed: group A (no.=11): GHRH (GHRH 1 29, Geref, Serono; 1 microg/kg iv at 150 min) preceded by saline or GHRH at 0 min; group B (no.=6): GHRH preceded by saline or rhGH (0.005 IU/kg iv at 0 min); group C (no.=6): GHRH preceded by rhGH alone or combined with GHRH; group D (no.=6): HEX (2 microg/kg iv at 150 min) alone or preceded by rhGH. In group A, the GH response to GHRH was not modified by pre-treatment with GHRH (GH peak, mean+/-SEM: 16.7+/-2.9 vs 15.1+/-2.3 microg/l, respectively). In group B, the GH response to GHRH was clearly inhibited by rhGH (8.7+/-2.3 vs 38.8+/-4.5 microg/l, p<0.001); the GH rise after rhGH in group B overlapped with that after GHRH in group A. In group C, the GH response to GHRH after pre-treatment with rhGH (13.2+/-4.0 microg/l) was similar to that in group B and was not significantly modified by pre-treatment with rhGH+ GHRH (6.9+/-2.7 microg/l); the GH rise after rhGH+GHRH was higher (p<0.05) than that after rhGH alone. In group D, the GH response to HEX was significantly blunted by pre-treatment with rhGH (34.1+/-11.7 vs 51.2+/-17.9 microg/l, p<0.05). Our results demonstrate that in childhood the somatotroph response to GHRH is preserved after GHRH while it is inhibited after rhGH administration, which is also able to blunt the GH response to HEX. Thus, the somatostatin-mediated negative GH auto-feedback is already operative in childhood; the reason why the GHRH- induced GH rise is not inhibited by GHRH pre treatment is unexplained. PMID- 10803473 TI - Repetitive graded ACTH stimulation test for adrenal insufficiency. AB - ACTH stimulation is useful in assessing the hypothalamo-pituitary-adrenal axis. However, there is still some debate about the proper dose and interpretation. We designed a new protocol using repetitive graded ACTH stimulation. Thirty-two patients with the diagnosis of adrenal insufficiency (Al) were studied. After taking samples for baseline ACTH and cortisol, 1 microg fresh-prepared ACTH (Synacthen (1-24)) was injected intravenously, then 5, 50 and 100 microg at hourly interval. Cortisol responses were measured at 30, 60, 120, 180 and 240 min. The secondary Al group (26 subjects) had cortisol responses in between those of the control group (8 subjects) and the primary Al group (6 subjects). The minimal overlap between the secondary Al group and the control group occurred at a 30-min cortisol response after 1 microg ACTH stimulation, using 20 microg/dl as the cut-off level. There was only one exception which showed an episodic release at 30 min. There were 5, 10 and 9 patients with secondary Al who responded normally to 5, 50 and 100 microg ACTH stimulation, respectively. Maximal cortisol increments of the primary Al group were all below 4 microg/dl. Although there were 11 cases of secondary Al whose cortisol responses overlapped with those of primary Al, only two of them had a cortisol increment less than 4 microg/dl. Our new protocol combines the advantage of the low dose ACTH stimulation test, a sensitive method for detecting mild Al, and the ACTH infusion test, a longer test to mimic surgical stress. PMID- 10803474 TI - Pendrin does not increase sulfate uptake in mammalian COS-7 cells. AB - Pendred's syndrome is characterized by goiter, sensorineural deafness and impaired iodide organification. It is one of the most frequent causes of congenital deafness accounting for about 10% of hereditary hearing loss. It is caused by mutations in the pendrin (PDS) gene, which was postulated to be a sulfate transporter, because of its homology with other genes. We tested sulfate transport in mammalian COS-7 cells that were transiently transfected with PDS cDNA. 35SO4 uptake increased in a time-dependent manner, but this phenomenon was similar in cells transfected with PDS and in mock-transfected cells (450 and 360 cpm/beta-gal units at 10 min, respectively; 38,250 and 31,000 cpm/beta-gal units, at 12 h, respectively). There was no significant increase in 35SO4 uptake using increasing amounts of PDS-containing plasmid (up to 12 microg per dish). These data indicate that pendrin is not a sulfate transporter. Additional functional studies on this protein are warranted to clarify its role in thyroid pathophysiology and inner ear development. PMID- 10803475 TI - Sunlight exposure and vitamin D deficiency in Turkish women. AB - Vitamin D is an essential steroid involved in bone metabolism, cell growth, differentiation, and regulation of the minerals in the body. The main sources of this vital vitamin are adequate diet and photosynthesis in the skin. The aim of this study was to investigate the efficiency of vitamin D synthesis in 48 premenopausal women (14-44 years) in relation to three different types of dressing in summer. Women in the first group (Group I) dressed in a style which exposed the usual areas of the skin to sunlight; women in the second group (Group II wore traditional clothing with the skin of the hands and face uncovered, while the third group (Group III) dressed in traditional Islamic style, covering the whole body including hands and face. Serum 25OHD levels of Group I, Group II, and Group III were 56+/-41.3 nmol/l, 31.9+/-24.4 nmol/l, 9+/-5.7 nmol/l, respectively (Group I vs Group III, p<0.001; Group II vs Group III, p<0,03; Group I vs Group II, p>0.05). Vitamin D levels were low in 44 percent of the Group I and 60% of the Group II, which suggested that sun exposure of skin areas of hands and face may partially provide vitamin D synthesis, but may not be enough to eliminate vitamin D deficiency. All the patients in group III had vitamin D levels below normal. This study emphasizes the necessity of vitamin D fortification of food even in a sunny country where some people may not be exposed to sunlight because of inappropriate clothing or an indoor-life. PMID- 10803476 TI - Dexamethasone effects on cortisol secretion in Alzheimer's disease: some clinical and hormonal features in suppressor and nonsuppressor patients. AB - Alterations in the hypothalamic-pituitary-adrenal axis (HPAA) and failure of dexamethasone (DXT) to suppress cortisol secretion occur in Alzheimer's disease (AD). This study was aimed to settle possible differences in some clinical (age, body weight, body mass index, dementia severity) and hormonal parameters in AD patients non-responders to overnight 1 mg-DXT suppression test compared with the responder subjects. ACTH, cortisol and dehydroepiandrosterone sulphate (DHEAS) day-time levels were assessed in 25 AD patients and in 12 age-matched healthy controls before DXT administration. In view of their neuroprotective effects, plasma levels of Insulin-like Growth Factor-I (IGF-I) and of IGF-Binding Proteins (IGFBPs) were also determined. After DXT, 8 AD subjects (32%) showed cortisol levels above the conventional cut-off of 140 nmol/L. No significant differences were found in clinical parameters in suppressor vs nonsuppressor patients. AD subjects showed higher cortisol, cortisol/DHEAS ratios, and lower DHEAS levels in comparison with controls. Both ACTH and cortisol levels were not different in suppressor and nonsuppressor patients, but DHEAS levels were significantly lower in nonsuppressor cases, who also exhibited ACTH and cortisol periodicities more altered than in suppressor and in control subjects. IGF-I and IGFBP-3 levels were lower and those of IGFBP-1 higher in nonsuppressor than in suppressor cases and in healthy controls. IGF-I/IGFBPs system data were correlated with cognitive impairment and adrenal steroid levels in AD patients. PMID- 10803477 TI - Natural course of insulin edema. AB - Generalized edema due to water retention is a very rare complication of insulin therapy. It affects mainly patients with newly diagnosed diabetes or patients with chronic hyperglycemia following initiation of insulin therapy. When it occurs, it is treated effectively with diuretics. This case report describes a female patient, who developed severe insulin edema following initiation of insulin. Diuretics were not given due to severe side effects, thus the natural outcome of insulin edema was observed. Edema was gradually replaced by fat tissue with persistent weight gain. Physicians treating diabetic patients should be aware of "insulin edema" in the differential diagnosis of weight gain in patients treated with insulin. PMID- 10803478 TI - A rare pituitary lesion. AB - The majority of the sellar masses are pituitary adenomas. Non-adenomatous hypophyseal lesions are rare and include lymphocytic hypophysitis, idiopathic giant cell hypophysitis and granulomatous hypophysitis. The latter include lesions caused by conditions like tuberculosis, sarcoidosis, syphilis, histiocytosis X and mycotic infections. We report a case of a Sudanese woman who presented with a pituitary mass (presumed preoperatively to be an adenoma) and underwent trans-sphenoidal surgery. The final histopathology showed tuberculous hypophysitis. The patient did not have history of prior exposure to tuberculosis nor any evidence of infection elsewhere in the body. We also briefly review the literature on pituitary tuberculomas. PMID- 10803479 TI - Inferior petrosal sinus sampling: pros and cons; when and where. PMID- 10803481 TI - Liver dysfunction associated with congenital hypopituitarism. PMID- 10803482 TI - "The dwarf'. Giacomo A.M. Ceruti, Milano, Italy (1698-1767). PMID- 10803480 TI - Testosterone therapy in men: clinical and pharmacological perspectives. AB - There is increasing evidence that androgen therapy in men may be effectively applied in several conditions to improve well being and health. Classical indications for androgen therapy in males are represented by primary or secondary hypogonadism, delayed puberty, aplastic anemia and that secondary to chronic renal failure, protein wasting diseases such as trauma, burns, tumors and infectious diseases. Androgen innovating applications in men are represented by aging and visceral obesity associated with the metabolic syndrome. In addition, it is clear that appropriate testosterone treatment can be adequately used in male contraception, provided spermatogenesis is abolished and tolerability is adequate. Due to unphysiological hormone levels achieved by currently available testosterone preparations, new delivery systems have been produced to achieve more physiological and sustained hormone levels and improve tolerability and action at the levels of target tissues. Some of them are now available in several countries and new formulas are under development. PMID- 10803483 TI - Awareness, management and follow-up of lipoprotein disturbances in hypertension. PMID- 10803484 TI - Hypertension: an important precursor of heart failure. AB - Hypertension is an important risk factor for cardiovascular morbidity and mortality. Hypertension is associated with the development of congestive heart failure by way of left ventricular hypertrophy, with left ventricular dilatation and through myocardial ischemia and left ventricular damage. Reports on the natural history of untreated hypertension indicate that at least 50% of affected subjects develop congestive heart failure. Hypertension is an important precursor of heart failure, and still the most common risk factor for congestive heart failure in the population. In clinical trials, particularly in elderly patients, a reduction in the incidence of congestive heart failure has been observed. Despite increments in the use of antihypertensive drugs, mortality from congestive heart failure among the elderly is increasing. Moreover, several patients with hypertension are unaware, untreated and uncontrolled for the most important risk factor for congestive heart failure. For the primary prevention of heart failure, improvements in blood pressure control are of vast importance. PMID- 10803485 TI - Beta-blocking agents in patients with insulin resistance: effects of vasodilating beta-blockers. AB - Essential hypertension is--at least in many subjects--associated with a decrease in insulin sensitivity, while glycaemic control is (still) normal. It seems that in hypertensive patients, two major functions of insulin are impaired: there is insulin resistance of peripheral glucose uptake (primarily skeletal muscle) and insulin resistance of insulin-stimulated vasodilation. In view of some retrospective data and meta-analyses, which showed a less than expected reduction in coronary events (coronary paradox), the metabolic side effects of the antihypertensive treatment have received more attention. Many groups have shown that conventional antihypertensive treatment, both with beta-blockers and/or diuretics, decreases insulin sensitivity by various mechanisms. While low-dose diuretics seem to be free of these metabolic effects, there is no evidence for this in the beta-adrenergic blockers. However, recent metabolic studies evaluated the effects of vasodilating beta-blockers, such as dilevalol, carvedilol and celiprolol, on insulin sensitivity and the atherogenic risk factors. None of them decreased insulin sensitivity, as has been described for the beta-blockers with and without beta1 selectivity. This supports the idea that peripheral vascular resistance and peripheral blood flow play a central role in mediating the metabolic side effects of the beta-blocking agents, as the vasodilating action (either via beta2 stimulation or alpha1-blockade) seems to more than offset the detrimental effects of the blockade of beta (or beta1) receptors. Further studies are needed to elucidate the relevance of the radical scavenging properties of these agents and their connection to their metabolic effects. Therefore, the beneficial characteristics of these newer beta-adrenoreceptor blockers suggest that the vasodilating beta-blocking agents could be advantageous for hypertensive patients with insulin resistance or type 2 diabetes. PMID- 10803486 TI - Effects of fosinopril treatment on blood pressure during physical and mental stress test in essential hypertension. AB - In this study the effects of once daily administration of 10 mg fosinopril on blood pressure response to mental and physical stress were evaluated in mild to moderate essential hypertension. A total of 25 patients (14 women, 9 men) with mild to moderate essential hypertension were enrolled in this study. Before the treatment and after 10 mg/day fosinopril treatment for 2 months, systolic and diastolic blood pressure and heart rate were recorded at rest and while performing a mental arithmetic test. In addition, exercise stress testing with Bruce protocol were performed before and after the treatment and systolic and diastolic blood pressure and heart rate were recorded at rest and during peak exercise. Statistical analysis were done by using the paired t-test. During the mental arithmetic test, systolic and diastolic blood pressure were significantly reduced (p < 0.005 and p < 0.001, respectively) after 2 months fosinopril treatment. Systolic and diastolic blood pressure were also significantly reduced during the exercise stress test (p < 0.005 and p < 0.05) after the treatment. Heart rate did not change during either the mental arithmetic or the exercise test. As a result, once daily 10 mg fosinopril may be effective in reducing blood pressure, not only at rest, but also during stressful situations. PMID- 10803487 TI - Cholesterol levels in untreated Spanish hypertensive patients. The Compas Study Group, Spanish Hypertension Society. AB - In daily practice, arterial hypertension (AHT) and hypercholesterolaemia are frequently associated with the existence of multiple common etiopathogenic interrelationships. This situation leads to an exponential increase in cardiovascular risk for these patients, so it is essential to know the prevalence and therapeutic management of hypercholesterolaemia in the hypertensive patient. This national study analyses the distribution of total cholesterol levels and low density lipoprotein cholesterol as well as hypercholesterolaemia prevalence and its therapeutic management in the uncontrolled hypertensive Spanish population. We observed mean total cholesterol levels of 227+/-41 mg/dl with a high prevalence of hypercholesterolaemia (34.2%) among hypertensive patients, and the percentage of those patients with "desirable" total cholesterol levels (<200 mg/dl) was <25%. The treated hypertensive patients presented both significantly higher mean cholesterol levels and greater hypercholesterolaemia prevalence than the untreated hypertensive patients. It appears that total cholesterol levels are scarcely related to the presence or non-presence of obesity, diabetes or smoking. Regarding treatment, only 14.6% of the hypercholesterolaemic hypertensive patients received hypolipaemic treatment with statins. These results support the need to introduce measures for better diagnostic and therapeutic management of hypercholesterolaemic hypertensive patients that will lead to a much higher reduction in cardiovascular risk for these patients. PMID- 10803488 TI - Prostacyclin: its pathogenic role in essential hypertension and the class effect of ACE inhibitors on prostaglandin metabolism. AB - Angiotensin-converting enzyme inhibitors (ACEI) block degradation of bradykinin, and bradykinin stimulates prostacyclin synthesis. Therefore, we set out to determine whether the effects of ACE inhibitors on prostaglandin production in essential hypertensive patients are class effects or are dependent on ACE inhibitor structure. In addition, we studied whether hypertensives show an impaired capacity to synthesize vasodilator prostaglandins. To address these questions, we compared the effects of captopril (sulfhydryl-containing inhibitor), enalapril and ramipril (carboxyl-containing inhibitors) and fosinopril (phosphoryl-containing inhibitor) on blood pressure and urinary excretion of 6-keto-prostaglandin (PG) F1-alpha (the breakdown product of prostacyclin) in 44 mild-to-moderate essential hypertensive subjects before and 8 weeks after administration of an ACEI. We also studied prostacyclin excretion in 15 normotensive healthy controls. Levels of urinary 6-keto-PGF1-alpha (pg/ml) were measured by specific radioimmunoassay. Hypertensive subjects showed a lower excretion of 6-keto-PGF1-alpha than normotensive controls (212+/-147 vs 353+/-98 pg/ml, p < 0.001). All ACEI induced a significant decrease in MAP and increased the rate of excretion of the prostacyclin metabolite: C, 211+/-200 to 338+/-250 pg/ml, p < 0.05; E, 202+/-133 to 296+/-207 pg/ml, p < 0.05; R, 205+/-127 to 342+/ 211 pg/ml, p < 0.05; F, 235+/-128 to 347+/-241 pg/ml, p < 0.05. In hypertensives (n = 44) the decrease in blood pressure correlated negatively with the rise in 6 keto-PGF1-alpha excretion (r = -0.51, p < 0.001). These data suggest that impaired prostacyclin biosynthesis in hypertensive patients could account for haemodynamic changes leading to the hypertensive state. Moreover, the hypotensive mechanisms of ACEI may be mediated by an increase in prostacyclin production; this effect seems to be class-dependent. PMID- 10803489 TI - Catecholamine storage vesicle protein expression in genetic hypertension. AB - Chromogranin A expression is heritable in humans, and both plasma chromogranin A concentration and its releasable adrenal and sympathetic neuronal pools are augmented in established essential (hereditary) hypertension. To evaluate chromogranin A further as a simpler or "intermediate phenotype" in the complex trait of hypertension, we studied chromogranin A expression in the spontaneously hypertensive rat (SHR), a rodent model of essential hypertension. Both plasma (p < 0.0001) and adrenal medullary (p = 0.003 to p < 0.0001) chromogranin A were elevated in the SHR, even at the earliest stages (3-4 weeks of age). In the adult adrenal gland, both chromogranin A (p=0.005) and norepinephrine (p=0.011) were increased in the SHR, while dopamine beta-hydroxylase activity was diminished (p < 0.0001). Chromogranin A mRNA expression was also elevated in the SHR adrenal medulla (p = 0.017). Differences in chromogranin A processing were not noted between SHR and Wistar Kyoto control (WKY) rats. In an SHR x WKY genetic intercross, control of the adrenal chromogranin A phenotype by a single major locus was suggested by comparison of phenotypic variance of the F2 vs F1 generations, and by bimodal frequency histogram (3:1 ratio), confirmed by maximum likelihood analysis (chi2 = 74.6, p < 0.000001) in the F2 generation. However, microsatellite alleles at a surrogate locus (Ighe) 12.7 cM from chromogranin A (Chga), on rat chromosome 6, failed to co-segregate with blood pressure in an F2 generation (F = 0.06, p = 0.94). In another rodent model of hereditary hypertension, the genetically hypertensive mouse (BPH/2), adrenal chromogranin A (p=0.018) and norepinephrine (p = 0.004) were actually diminished. We conclude that over-expression of chromogranin A is a variable feature of mammalian genetic hypertension. In one rodent model (the SHR), over-expression of chromogranin A is largely controlled by a single genetic locus, but the chromogranin A locus itself is not directly linked to determination of the blood pressure elevation of the SHR. PMID- 10803490 TI - Progress report on the Nordic diltiazem study (NORDIL): an outcome study in hypertensive patients. AB - NORDIL--the Nordic Diltiazem Study (NORDIL)--is a prospective, randomized, open blinded-endpoint (PROBE), multicenter, parallel-group morbidity/mortality outcome study in hypertensive patients designed to compare an intervention strategy based on the calcium antagonist diltiazem with a strategy based on conventional antihypertensive drug treatment (diuretics or beta-adrenergic blockers). Patient recruitment was started in Norway and Sweden in September 1992, and ended on December 15, 1996, when 10.896 male and female patients, aged 50-74 years, with essential hypertension had been randomized. In this paper we describe the baseline data of the patient cohort and blood pressures achieved in the two treatment groups during the early part of the study. The patient cohort consists of 5294 males and 5602 females with a mean age of 59.6 and 60.3 years, respectively. Concomitant disorders and risk factors in the cohort are: smoking 22%, ischemic heart disease 3.0%, previous myocardial infarction (MI) 2.0%, previous stroke 1.5%, diabetes mellitus 7.0%, and renal impairment 0.3%. There were no differences between the treatment groups in these respects. The blood pressure treatment goal is a target diastolic blood pressure of < or =90 mmHg or a 10% diastolic blood pressure reduction from the inclusion pressure. In the treatment group randomized to a diltiazem-based treatment strategy, blood pressure was 174/106 mmHg at baseline and 156/90 mmHg after 12 months of follow up on active treatment. In the group randomized to a conventional treatment strategy, baseline blood pressure at randomization was 173/106 mmHg and 153/90 mmHg after 12 months on active therapy. The NORDIL study will terminate on October 31, 1999 and the final results should be available by mid-2000. PMID- 10803491 TI - Neonatal sympathectomy reduces adult blood pressure and cardiovascular pathology in Y chromosome consomic rats. AB - The hypothesis was tested that the sympathetic nervous system (SNS) developmentally influences circulating testosterone (T), systolic blood pressure (SBP) and cardio-renal pathology in SHR/y animals. A sympathoplegic drug, guanethidine, and an antibody to nerve growth factor were administered to WKY and borderline hypertensive SHR/y male rats (n = 20/group) for the first 3 weeks of life; control groups (n = 20/group) received saline. SBP, serum T and luteinizing hormone (LH) were measured. SBP in the WKY and SHR/y sympathectomy (sympx) groups decreased 10mmHg (p < 0.001) and 50mmHg (p < 0.001), respectively, when compared to their control groups. Serum T levels in the sympx WKY group were lower (p < 0.01) than those in controls, and the rise of T typically observed in SHR/y from weeks 6-8 was delayed in the sympx SHR/y group, similar to the pattern in WKY. Serum LH levels were increased in the sympx WKY group, but not in the SHR/y group. Sympx caused a greater reduction in renal glomerular changes (p < 0.01), coronary artery collagen deposition (p < 0.01) and myocardial fibrosis (p < 0.01) in SHR/y than WKY rats. In conclusion, the SHR Y chromosome has a locus that enhances SNS activity, which can raise SBP and result in renal and cardiovascular tissue damage. PMID- 10803492 TI - Augmentation of BNP gene expression in atria by pressure overload in transgenic rats harbouring human renin and angiotensinogen genes. AB - We studied the role of angiotensin II in pressure overload-induced B-type natriuretic peptide (BNP) gene expression by using a double transgenic rat (dTGR) model, in which transgenic rats for the human angiotensinogen and renin genes are crossed. Pressure overload produced by [Arg8]-vasopressin (AVP) infusion (i.v., 0.05 microg/kg/min for 2 h) in conscious, chronically instrumented rats, resulted in a significantly greater increase in BNP mRNA levels in the left atrium of the dTGR rats than in Sprague-Dawley (SD) control rats (3.6- vs 1.6-fold, p < 0.05), while in the left ventricle there was no significant difference between the strains. In dTGR rats, the early activation of the BNP gene expression was associated with a decrease in immunoreactive BNP levels in the atrium (27.5%, p < 0.05), but not in the ventricle. In SD rats, ir-BNP levels did not change significantly in either atria or ventricles in response to AVP infusion. These results show that the pressure overload-induced activation of BNP gene expression differs between atrial and ventricular myocytes in the dTGR model of experimental hypertension. PMID- 10803493 TI - Agonist-like activity of antibodies to angiotensin II receptor subtype 1 (AT1) from rats immunized with AT1 receptor peptide. AB - In the present study, rats were immunized with angiotensin II receptor subtype 1 (AT1) receptor peptides for 3 months to see if the immunization produced specific anti-AT1 receptor antibodies and if continuous stimulation for 3 months affected blood pressure or induced morphological changes in the organs containing AT1 receptors. Our results showed that there were constant high levels of circulating antibodies throughout the study period in all rats of the immunized group, but not in the control rats, and that there were almost no significant cross reactions of antisera with AT2 receptor peptide and alpha1 adrenoceptor peptide, except in four rats, which showed low cross-reactions with alpha1 adrenoceptor and AT2 receptor peptides. When an affinity-purified anti-AT1 receptor antibody was used, it specifically displayed the AT1-stimulatory positive chronotropic effect and also localized AT1 receptors. However, in the immunized group, saturation binding of AT1 in homogenates from kidneys showed no difference either in maximal binding sites (Bmax) or in antagonist affinity (Kd). No difference in mRNA of AT1a was found in either kidney or heart, and no morphological changes in the organs were observed, as compared with the control group. Furthermore, immunization did not cause hypertension. In conclusion, the synthetic peptide corresponding to the second extra-cellular loop of the human AT1 receptor was able to produce highly specific and functionally active anti-AT1 receptor antibodies, but unable to induce pathological structural changes or hypertension. PMID- 10803494 TI - Characterization and distribution of a new enterotoxin-related superantigen produced by Staphylococcus aureus. AB - Staphylococcal enterotoxins (SEs) are a family of structurally related pyrogenic exotoxins consisting of the five prototypic SEs (types A to E) and three newly characterized SEs (types G to I) produced by Staphylococcus aureus (S. aureus). They also work as superantigens and cause food poisoning and shock symptoms in humans. In this study, we cloned a new variant gene of the seg and characterized its superantigenic properties and distribution among the clinical isolates of S. aureus. The gene encodes a 233 amino acid protein which is highly homologous to SEG (97.7%). The variant SEG (SEGv) expressed by the cloned gene exerted mitogenic activity on human peripheral blood mononuclear cells at the concentration of 100 pg/ml. T cells bearing Vbeta3, 12, 13.1, 13.2, 14 and 15 were preferentially expanded after stimulation with the recombinant protein. The mRNA of the variant seg gene was detected in the total RNA of the organisms bearing this gene. By PCR, 27 out of 48 clinical isolates of S. aureus (56%) possessed either the seg or variant seg gene. These findings suggest that SEG, or SEGv, is one of the most frequently produced superantigen exotoxins by S. aureus and may participate in the inflammatory process of the host by activating a distinct set of Vbeta families of T cells. PMID- 10803495 TI - Effect of antibiotics, levofloxacin and fosfomycin, on a mouse model with Escherichia coli O157 infection. AB - There have been some reservations about the treatment of enterohemorrhagic Escherichia coli (EHEC) infection with antibiotics to prevent the occurrence of hemolytic uremic syndrome (HUS). However, the administration of antimicrobial agents for EHEC infection is under discussion. Therefore, we used an experimental mouse model to assess the advantage/disadvantage of two major antibiotics, levofloxacin (LVFX) and fosfomycin (FOM). Germ-free IQI mice were inoculated with EHEC O157 strain EDL931 or #7. Bacteria colonized feces at 10(9)-10(10) CFU/g, and Shiga toxins (STXs) were detected in the feces. From 1 day after infection, mice were assigned to LVFX (20 mg/kg) once daily or FOM (400 mg/kg) once daily. A significant decrease in overall mortality was observed after treatment of LVFX, with EHEC disappearing immediately from the feces of mice. FOM also reduced mortality for one strain, the STX level decreased gradually. LVFX exhibited higher therapeutic efficacy than FOM. Strain differences were observed in the model during the treatment. PMID- 10803496 TI - Staphylococcus aureus produces autolysin-susceptible cell walls during growth in a high-NaCl and low-Ca2+ concentration medium. AB - The growth of Staphylococcus aureus 209P becomes unusually sensitive to a high NaCl concentration by decreasing the Ca2+ concentration in growth media, and cells either autolyze or transform into protoplast-like forms when grown standing in high-NaCl and low-Ca2+ concentration media below 37 C (Ochiai, T., Microbiol. Immunol. 43 (7): 705-709, 1999). To assess the role of Ca2+ in the salt tolerance of this organism, cells grown in the presence of different concentrations of Ca2+ were treated with boiling SDS, and their susceptibilities to crude autolysin (3 M LiCl extract of S. aureus 209P cells) were evaluated by turbidimetric assay and zymographic analysis. Susceptibilities of SDS-treated cells (SDS-cells) to crude autolysin were significantly influenced by Ca2+ concentration in the culture, and SDS-cells prepared from cultures grown in high-NaCl and high-Ca2+ concentration media exhibited marked resistance to crude autolysin when the assay system contained a high concentration of NaCl. On the contrary, SDS-cells prepared from cultures grown in high-NaCl and low-Ca2+ concentration media were rather susceptible to crude autolysin under the same assay conditions. A zymographic analysis revealed that the constitution of cell-associated autolysins was not influenced by the concentration of exogenous Ca2+. These results suggested that at least part of the mechanism of salt-induced autolysis in S. aureus 209P might be related to the synthesis of an autolysin susceptible cell wall. PMID- 10803497 TI - Induction of high levels of epitope-specific antibodies by epitope/peptide candidate vaccines against human immunodeficiency virus type-1 (HIV-1). AB - To test the immunogenicity of GPGRAFY-epitope-based candidate vaccines, a peptide with four repetitive GPGRAFY epitopes, V3-P1 [C-(GPGRAFY)4], and a peptide (PND) of the principal neutralizing domain (V3 loop: amino acid 301-328: C TRPNNNTRKSIRIQRGPGRAFYTIGKI) on gp120 were synthesized and covalently coupled to a carrier protein BSA. Immunization of BALB/c mice and New Zealand White Rabbits with these conjugate vaccines engendered strong antibody responses against the PND (mouse serum titer by 1:12,800-25,600; rabbit serum titer by 1:6,400-12,800). Interestingly, the V3-P1-BSA conjugates and the PND-BSA conjugates could induce high levels of GPGRAFY-epitope-specific antibodies in the mice and rabbits (mouse serum titer by 1:25,600; rabbit serum titer by 1:12,800-25,600), while a recombinant gp160 subunit vaccine induced a low level of GPGRAFY-epitope-specific antibodies (serum titer by 1:400-1,600 in mice and rabbits). To confirm the above results, GPGRAFY-epitope-specific antibodies were isolated from rabbit sera induced by V3-P1-BSA, PND-BSA conjugates and rgp160 vaccine. In fact, 23-38 and 13-22 microg epitope-specific antibodies per milliliter serum were isolated from rabbit sera induced by V3-P1-BSA and PND-BSA conjugate, respectively, while 1.34 microg epitope-specific antibodies per milliliter serum were identified in rabbit serum induced by rgp160 vaccine. In the control group, only 0.069 microg proteins per milliliter serum were found in pooled pre-immune serum (normal serum). These results from mouse and rabbit experiments indicate that epitope and peptide vaccines both induce high levels of GPGRAFY-epitope-specific antibodies in comparison with rgp160 subunit vaccine, suggesting that epitope/peptide vaccines may be a new strategy to induce protective activity. PMID- 10803498 TI - Exposure of normal monocyte-derived dendritic cells to human immunodeficiency virus type-1 particles leads to the induction of apoptosis in co-cultured CD4+ as well as CD8+ T cells. AB - The depletion of immune T cells by human immunodeficiency virus type-1 (HIV-1) infection is a major mechanism involved in the pathogenesis of AIDS. Here, we examined a possible effector function of blood monocyte-derived dendritic cells (DCs) to induce apoptosis in bystander CD4+ and CD8+ T cells. The DCs were generated by culturing monocytes in the presence of granulocyte-macrophage colony stimulating factor and interleukin-4. The DCs exposed to HIV-1 particles were co cultured with healthy donor-derived blood T cells at a ratio of 1:20. Analyses by percent cell mortality, staining with propidium iodide and reactivity with Annexin V revealed the induction of apoptosis in both CD4+ and CD8+ target T cells. Further, this apoptosis occurred without stimulation with mitogens when the cell cycle of target T cells shifted from G0 to G1, probably due to the mitogenic effect of the DCs. Thus, induction of apoptosis in both CD4+ and CD8+ T cells occurred via interaction with DCs adsorbed with HIV-1 particles. PMID- 10803499 TI - Efficient induction of peptide-specific cytotoxic T lymphocytes by LPS-activated spleen cells. AB - Lipopolysaccharides of gram-negative bacteria are potent activators of B cells, dendritic cells and monocytes/macrophages. We have investigated the use of LPS activated spleen cells as antigen-presenting cells to induce CD8+ cytotoxic T lymphocytes in vivo that are reactive to MHC class I binding peptides. Compared with resting spleen cells, CTL induction was more efficient and less variable for different peptides with LPS-activated spleen cells. Cytotoxic responses were specific for the immunized peptides and contained high affinity CD8+ T cells. The removal of dendritic cells and monocytes/macrophages by Sephadex G10 column did not show profound effects on CTL induction, indicating that B-cell blasts were largely responsible. This easily accessible method should facilitate the screening of MHC class I binding peptides to determine whether or not the host's T-cell repertoire contains reactive T cells. PMID- 10803500 TI - SDF-1 has costimulatory effects on human T cells: possible involvement of MAPK (ERK2) activation. AB - Stromal cell-derived factor-1 (SDF-1) is an efficacious chemoattractant for lymphocytes, monocytes and hematopoietic progenitor cells. In the present study, we examined whether SDF-1 has growth promoting activity on human peripheral T cells and analyzed the possible underlying signal transduction pathways. SDF-1 augmented the proliferation of anti-CD3- or PHA-stimulated normal human PBMC in a dose-dependent manner but not that of resting PBMC. It was noted that SDF-1 alone could induce a significant proliferation of PHA-preactivated T cells. Anti-SDF-1 sera could inhibit the augmentation of T-cell proliferation in each experiment. Furthermore, Western blot analysis revealed that mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase 2 (ERK2), but not c-Jun N-terminal kinase (JNK) was activated by SDF-1. Considering that costimulatory signals have been reported to involve ERK2 activation, these results indicate that SDF-1 has costimulatory effects on T cells that are possibly mediated by ERK2 activation and may play a role in not only migration but also the potentiation or maintenance of T cells. PMID- 10803501 TI - Molecular characterization and functional analysis of a secA gene homolog in Actinobacillus actinomycetemcomitans. AB - Results of Southern blot analyses and polymerase chain reaction revealed that the Gram-negative pathogen, Actinobacillus actinomycetemcomitans, harbored DNA homologous to the secA gene of Escherichia coli. In E. coli, the secA gene product is essential for translocation of proteins across the inner membrane via the Sec system. This A. actinomycetemcomitans secA homolog was cloned and its nucleotide sequence determined. Amino acid sequence analysis of the cloned gene revealed significant homology to the SecA proteins of Haemophilus influenzae, E. coli, Caulobacter crescentus and Bacillus subtilis. Although the cloned gene did not complement a temperature sensitive mutation in the E. coli secA gene, strains harboring the cloned gene did produce a protein that cross-reacted with anti-SecA antibody. In addition, the cloned gene did restore sensitivity to sodium azide in an E. coli azide mutant. These data support the hypothesis that A. actinomycetemcomitans may use a system similar to the Sec system of E. coli to transport proteins across the cytoplasmic membrane, but suggest that the A. actinomycetemcomitans gene product may require genera-specific Sec proteins to complement some Sec mutations in E. coli. PMID- 10803502 TI - Analysis of Vibrio mimicus clinical strains by arbitrarily primed polymerase chain reaction. AB - A total of 51 Vibrio mimicus clinical strains from different geographic locations were examined by arbitrarily primed polymerase chain reaction (AP-PCR). The primer VMH-3 divided them into 28 groups, although 18 groups consisted of a single strain at present. All groups had a common 1.0-kb amplification fragment. Most of the groups consisted of strains from same region, although two exceptional groups showed a few amplification fragments including strains from different regions. AP-PCR groups were not consistently associated with serogroups. AP-PCR is thought to be a valuable and easy method for the epidemiological study of V. mimicus. PMID- 10803503 TI - Biological and chemical terrorism: strategic plan for preparedness and response. Recommendations of the CDC Strategic Planning Workgroup. AB - The U.S. national civilian vulnerability to the deliberate use of biological and chemical agents has been highlighted by recognition of substantial biological weapons development programs and arsenals in foreign countries, attempts to acquire or possess biological agents by militants, and high-profile terrorist attacks. Evaluation of this vulnerability has focused on the role public health will have detecting and managing the probable covert biological terrorist incident with the realization that the U.S. local, state, and federal infrastructure is already strained as a result of other important public health problems. In partnership with representatives for local and state health departments, other federal agencies, and medical and public health professional associations, CDC has developed a strategic plan to address the deliberate dissemination of biological or chemical agents. The plan contains recommendations to reduce U.S. vulnerability to biological and chemical terrorism--preparedness planning, detection and surveillance, laboratory analysis, emergency response, and communication systems. Training and research are integral components for achieving these recommendations. Success of the plan hinges on strengthening the relationships between medical and public health professionals and on building new partnerships with emergency management, the military, and law enforcement professionals. PMID- 10803504 TI - CD34: to select or not to select? That is the question. AB - Recent evidence suggests that expression of CD34 on the cell membrane does not always correlate with stem cell activity. In the mouse, there is a highly quiescent population of stem cells that lacks CD34 expression, but has full reconstituting capacity. The current review addresses the discovery of a similar population of dormant CD34-negative human hematopoietic stem cells. This information casts some uncertainty on the benefits of CD34+ cell isolation for stem cell transplantation, until more is known about the novel CD34-negative stem cell population. Methods designed to achieve removal of specific mature blood cell lineages might prove to be most advantageous in the future. PMID- 10803505 TI - Malignant hematopoietic cell lines: in vitro models for the study of natural killer cell leukemia-lymphoma. AB - Malignancies involving natural killer (NK) cells are rare disorders. The complexity of NK cell-involving disorders has only recently been appreciated. Modern classifications discern immature (precursor) from mature NK cell leukemias lymphomas. Continuous NK leukemia-lymphoma cell lines represent important model systems to study these neoplasms. While there are a number of putative NK cell lines which are, however, either not characterized, not immortalized, non malignant, non-NK, or plain false cell lines, six bona fide malignant NK cell lines have been established and are sufficiently well characterized: HANK1, KHYG 1, NK-92, NKL, NK-YS and YT. Except for YT which was derived from a not further defined acute lymphoblastic lymphoma, these cell lines were established from patients with various NK cell malignancies. Five of the six cell lines are constitutively interleukin-2-dependent. Their immunoprofile is remarkably similar: CD1-, CD2+, surface CD3 (but cytoplasmic CD3epsilon+), CD4-, CD5-, CD7+, CD8-, CD16-, CD56+, CD57-, TCRalphabeta-, TCRgammadelta-, negative for B cell and myelomonocytic markers. The immunoglobulin heavy chain and T cell receptor genes are all in germline configuration. All six lines show complex chromosomal alterations, with both numerical and structural aberrations, attesting to their malignant and monoclonal nature. Functionally, these cells which contain azurophilic granules in their cytoplasm are nearly universally positive in NK activity assays. Three of five cell lines are Epstein-Barr virus-positive (type II latency). The composite data on these six cell lines allow for the operational definition of a typical malignant NK cell line profile. NK leukemia-lymphoma cell lines will prove invaluable for studies of normal and malignant NK cell biology. PMID- 10803506 TI - New definition of remission in childhood acute lymphoblastic leukemia. AB - The extent of clearance of leukemic cells from the blood or bone marrow during the early phase of therapy is an independent prognostic factor in acute lymphoblastic leukemia (ALL). Several methods are available to measure the minimal residual disease (MRD) remaining after initial intensive chemotherapy. The most promising are flow cytometric detection of aberrant immunophenotypes and polymerase chain reaction analysis of clonal antigen-receptor gene rearrangements. When applied together, these techniques enable one to monitor MRD in virtually all cases of ALL. Patients who achieve an 'immunologic' or 'molecular' remission (ie leukemic involvement of <0.01% of nucleated bone marrow cells at the end of remission induction therapy) are predicted to have a better clinical outcome than patients whose remission is defined solely by morphologic criteria. In studies to date, patients with MRD at a level of 10(-2) or more at the end of induction have fared almost as poorly as those with > or =5% blast cells in the bone marrow (ie induction failures). Sequential monitoring of MRD can improve the clinical utility of risk assessment still further. Additional studies are needed to determine the critical levels of MRD at various times of treatment and whether therapeutic intervention based on MRD findings can improve clinical outcome. PMID- 10803507 TI - An effective chemotherapeutic regimen for acute myeloid leukemia and myelodysplastic syndrome in children with Down's syndrome. AB - In recent pediatric collaborative studies of acute myeloid leukemia (AML), patients with Down's syndrome (DS) have better outcome than other patients when they were treated according to their intensive AML protocols. This may be attributed to enhanced sensitivity of DS AML cells to selected chemotherapeutic agents. We evaluated a less intensive chemotherapeutic regimen which was specifically designed for children with AML-DS. Remission induction chemotherapy consisted of daunorubicin (25 mg/m2/day for 2 days), cytosine arabinoside (100 mg/m2/day for 7 days), and etoposide (150 mg/m2/day for 3 days). Patients received one to seven courses of consolidation therapy of the same regimen. Thirty-three patients were enrolled on the study and their clinical, hematologic and immunophenotypic features were analyzed. Of the 33 patients, all were younger than 4 years and diagnosed as having acute megakaryoblastic leukemia or myelodysplastic syndrome. All patients achieved a complete remission and estimated 8 year event-free survival rate was 80+/-7%. Three patients relapsed and two died due to cardiac toxicity and one due to septic shock. The results of our study showed that patients with AML-DS constitute a unique biologic subgroup and should be treated according to a less intensive protocol designed for AML-DS. PMID- 10803508 TI - Slow disappearance of peripheral blood blasts is an adverse prognostic factor in childhood T cell acute lymphoblastic leukemia: a Pediatric Oncology Group study. AB - The rapidity of response to induction therapy is emerging as an important prognostic factor in children and adolescents with acute lymphoblastic leukemia (ALL). We studied the relationship between rapidity of reduction in peripheral blood blast count and treatment outcome in children with T cell ALL (T-ALL). Initial systemic chemotherapy included prednisone, vincristine, doxorubicin and cyclophosphamide. A Cox analysis evaluated the correlation between the length of time that the peripheral blood absolute blast count (ABC) remained above 1000/mm3 following the start of treatment and event-free survival (EFS). Data were available for 281 patients. Patients for whom the ABC remained >1000/mm3 for 3 or more days following administration of intensive therapy had an estimated 5-year EFS of 34.2% (s.e. = 7.2) vs 58.3% (3.5) for those whose ABC was <1000/mm3 within 0-2 days, with a hazard ratio (HR) of failure of 2.03 (95% CI = 1.35-3.06, P < 0.001) for the slower responding patients. Pre-treatment of some type (usually with prednisone) occurred in 128 patients (average duration 1.7 days). When this was accounted for, patients with an ABC >1000/mm3 for 5 or more days following the start of treatment of any kind had a HR for failure of 2.27 (95% CI = 1.38 3.72, P < 0.001) compared to those responding within 0-4 days. Inclusion of other clinical and biological factors in a multivariate analysis did not alter the prognostic importance of slower blast clearance. Pediatric patients with T-ALL who have a circulating blast count >1000/mm3 at diagnosis and a relatively slower response to initial treatment are at increased risk of treatment failure. Rapidity of response may therefore be a clinically useful prognostic factor for patients with T-ALL. PMID- 10803510 TI - Allelotype analysis of the myelodysplastic syndrome. AB - Myelodysplastic syndromes (MDS) are a group of clonal hematologic disorders found predominantly in the elderly. The molecular mechanisms underlying the development of MDS remain obscure. In order to begin to identify tumor suppressor genes involved in these disorders, we performed a detailed microsatellite allelotype of chromosomal deletions associated with MDS. DNAs from both bone marrow and peripheral blood of 32 MDS patients were studied using 84 highly informative microsatellite markers on all autosomal arms, excluding the short arms of the acrocentric chromosomes. A high percentage of loss of heterozygosity (LOH) was identified on chromosome 5q (40% of informative cases), 7q (45%), 17p (23%) and 20q (20%), which corresponds to the most common cytogenetic abnormalities reported in MDS. In addition, a high incidence of LOH (> or =20%) was observed on chromosomal arms which had not been previously reported including 1p (36%), 1q (35%), and 18q (23%). This extensive allelotype analysis focuses attention on several novel genomic regions that probably contain novel tumor suppressor genes whose loss of function contributes to the development of MDS. PMID- 10803509 TI - Screening for MLL tandem duplication in 387 unselected patients with AML identify a prognostically unfavorable subset of AML. AB - Partial tandem duplications of the MLL gene have been associated with trisomy 11 in acute myeloid leukemia (AML) and recently, have also been reported for karyotypically normal AML. In order to test the incidence and prognostic importance of this molecular marker, we have analyzed eight cases of AML with trisomy 11 and 387 unselected consecutive cases with AML for partial duplications of the MLL gene. Patients with normal karyotypes and those with various chromosome aberrations were included. De novo as well as secondary leukemias including all FAB subtypes were analyzed. Performing a one-step RT-PCR with 35 cycles using an exon 9 forward primer and an exon 3 reverse primer partial tandem duplications of the MLL gene were demonstrated in 3/8 (37.5%) patients with trisomy 11. In addition, 13/387 (3.4%) of unselected cases revealed a tandem duplication. Ten of these 13 cases were cytogenetically normal, the other three cases had < or =2 additional chromosomal alterations. Sequencing of the RT-PCR products of all 16 positive cases revealed fusions of MLL exon 9/exon 3 (e9/e3) (six cases), e10/e3 (three cases), e11/e3 (four cases) or combinations of differentially spliced e10/e3 and e11/e3 (three cases) transcripts. The duplications were confirmed by genomic long range PCR and Southern blot hybridization. Twelve cases with the MLL duplication were de novo myeloid leukemia, one was a secondary AML after MDS, three were therapy-related AML (t AML). Of the 16 MLL-duplication positive cases, seven were classified as FAB M2, two as M1, five as M4, one as M0, one as M5b. The mean age was 62.3 years for patients with MLL duplication vs 50.3 years for the control group. Of 15 adult patients, 12 received treatment. Of these, three were nonresponders, five had early relapse (< or =6 months), four relapsed between 7 and 12 months. Median survival and relapse-free interval of the MLL duplication positive group was significantly worse than those of an age-matched karyotypically normal control group. In conclusion, MLL tandem duplications (1) are less common than previously reported; (2) are preferentially observed in AML with normal karyotypes, but can also be found in the presence of chromosome alterations; (3) are not strongly associated with an FAB subtype; (4) were not observed with the prognostically favorable t(8;21), inv(16), and t(15;17), other recurrent translocations, or in complex karyotypes; and (5) identifies a subgroup of patients with an unfavorable prognosis. PMID- 10803511 TI - Identification of three subgroups of B cell chronic lymphocytic leukemia based upon mutations of BCL-6 and IgV genes. AB - Although B cell chronic lymphocytic leukemia (B-CLL) has been traditionally viewed as a tumor of virgin B cells, this notion has been recently questioned by data suggesting that a fraction of B-CLL derives from antigen experienced B cells. In order to further clarify the histogenetic derivation of this lymphoproliferation, we have analyzed the DNA sequences of the 5' non-coding region of BCL-6 proto-oncogene in 28 cases of B-CLL. Mutations of BCL-6 proto oncogene, a zinc finger transcription factor implicated in lymphoma development, represent a histogenetic marker of B cell transit through the germinal center (GC) and occur frequently in B cell malignancies derived from GC or post-GC B cells. For comparison, the same tumor panel was analyzed for somatic mutations of the rearranged immunoglobulin variable (IgV) genes, which are known to be acquired at the time of B cell transit through the GC. Sequence analyses of BCL-6 and IgV genes allowed the definition of three groups of B-CLL. Group I B-CLL displayed mutations of both BCL-6 and IgV genes (10/28; 36%). Group II B-CLL displayed mutated IgV genes, but a germline BCL-6 gene (5/28; 18%). Finally, group III B-CLL included the remaining cases (13/28; 46%) that were characterized by the absence of somatic mutations of both BCL-6 and IgV genes. Overall, the distribution of BCL-6 and IgV mutations in B-CLL reinforce the notion that this leukemia is histogenetically heterogeneous and that a substantial subgroup of these lymphoproliferations derives from post-germinal center B cells. PMID- 10803512 TI - BIOMED-1 concerted action report: flow cytometric characterization of CD7+ cell subsets in normal bone marrow as a basis for the diagnosis and follow-up of T cell acute lymphoblastic leukemia (T-ALL). AB - The European BIOMED-1 Concerted Action was initiated in 1994 to improve and standardize the flow cytometric detection of minimal residual disease (MRD) in acute leukemia (AL). Three different protocols were defined to identify the normal subsets of B, T and myeloid cells in bone marrow (BM), and were applied to the different types of AL in order to study aberrant immunophenotypes. Using sensitive acquisition methods ('live gate') T cell subsets in normal BM could be identified with five triple-stains: CD7/CD5/CD3, CD7/CD4/CD8, CD7/CD2/CD3, CD7/CD38/CD34 and TdT/CD7/surface or cytoplasmic (cy)CD3 (antibodies conjugated with FITC/PE/PECy5 or PerCP, respectively). The identification of T cell subsets in BM allowed definition of 'empty spaces' (ie areas of flow cytometric plots where normally no cells are found). All studied T-ALL cases (n = 65) were located in 'empty spaces' and could be discriminated from normal T cells. The most informative triple staining was TdT/CD7/cyCD3, which was aberrant in 91% of T-ALL cases. In most cases, two or more aberrant patterns were found. Apparently the immunophenotypes of T-ALL differ significantly from normal BM T cells. This is mostly caused by their thymocytic origin, but also the neoplastic transformation might have affected antigen expression patterns. Application of the five proposed marker combinations in T-ALL contributes to standardized detection of MRD, since cells persistent or reappearing in the 'empty spaces' can be easily identified in follow-up BM samples during and after treatment. PMID- 10803513 TI - Arginine deiminase inhibits proliferation of human leukemia cells more potently than asparaginase by inducing cell cycle arrest and apoptosis. AB - L-Asparaginase is used for the treatment of acute leukemias, but is sometimes ineffective or associated with severe side-effects. We report here that the enzyme arginine deiminase is approximately 100-fold more potent than L asparaginase in inhibiting the proliferation of cultured human lymphatic leukemia cell lines while it appears to be less effective in leukemia cells of myeloid origin. The inhibition of cell proliferation involves cell growth arrest in the G1- and/or S-phase and eventually apoptotic cell death. Our results suggest the possibility of a future use of arginine deiminase for the therapy of leukemia. PMID- 10803514 TI - Clonal variability in CD95 expression is the major determinant in Fas-medicated, but not chemotherapy-medicated apoptosis in the RPMI 8226 multiple myeloma cell line. AB - CD95 (Fas/APO-1) is a member of the TNFR superfamily that induces apoptosis following cross-linking with its cognate ligand, CD95L (FasL/APO-1L) or agonist antibody. The human myeloma cell line, RPMI 8226, has limited sensitivity to CD95 mediated apoptosis, with a maximum of 65% of the population responding. To determine the source of the limited sensitivity to CD95-mediated apoptosis, we isolated multiple clones from the RPMI-8226 cell line by limiting dilution. Analysis of these clones demonstrated that sensitivity to CD95-mediated cell death directly correlated with CD95 expression. Clones with high levels of CD95 expression had greater than 90% cell death, whereas cells with low levels of expression had less than 10% cell death. In contrast, no correlative differences were identified for other members of the DISC complex, or for members of the anti apoptotic Bcl-2 family. We further examined the sensitivity of the 8226 clones to various cytotoxic agents. Although modest clonal variability was demonstrated in response to the chemotherapeutic drugs, doxorubicin, etoposide (VP-16), and vincristine, there was no correlation between CD95 function and sensitivity to chemotherapeutic drugs. These results indicate that in this cell line, receptor expression is rate limiting in CD95-mediated apoptosis, whereas CD95 expression was not a determinant in drug-induced programmed cell death. PMID- 10803515 TI - Growth inhibition and synergistic induction of apoptosis by zoledronate and dexamethasone in human myeloma cell lines. AB - Bisphosphonates (BPs) are commonly used in the treatment of myeloma-associated osteolytic lesions. Recent reports have suggested that BPs may also exert direct antitumor effects on myeloma cells. Here, we show that the treatment of myeloma cell lines with the combination of the potent BP zoledronate and dexamethasone inhibits cell growth and synergistically induces apoptotic cell death, providing a rationale for potential applications in vivo. PMID- 10803516 TI - Inactivation of wild-type BCR/ABL tyrosine kinase in hematopoietic cells by mild hyperthermia. AB - Temperature-sensitive mutants of BCR/ABL tyrosine kinase have been extensively used to study the mechanisms of cell transformation and signal transduction. However, little is known about the effect of temperature on the activity of wild type BCR/ABL gene product. In this study, we demonstrate that in vivo tyrosine kinase activity of p210, p190 BCR/ABL and v-abl are temperature-sensitive when expressed in hematopoietic cells and decline when temperature is raised 2 degrees C above normal range. In vitro tyrosine kinase activities of purified recombinant Abl and immunoprecipitated p210 BCR/ABL were also sensitive to increased temperature. Tyrosine phosphorylation of cellular proteins was markedly reduced in BCR/ABL transformed cells after 16 h at 39 degrees C, whereas the expression of BCR/ABL was unchanged. Temperature-induced downregulation of BCR/ABL kinase activity was reversible when cells were shifted back to 37 degrees C. The downregulation of Abl tyrosine kinase activity was not influenced by mutation or deletion of SH2 or SH3 domains or mutation of the GRB2 binding site. No increase in functional activity or expression of protein-tyrosine phosphatases, PTP-1B, SH PTP1 or SH-PTP2 was detected in cells grown at 39 degrees C. Temperature-induced downregulation in tyrosine kinase activity correlated with decline in phosphotyrosine-associated PI 3-kinase whereas there was no change in growth factor independence of transformed hematopoietic cells. In conclusion, Abl tyrosine kinase has intrinsic sensitivity to temperature and BCR/ABL expressed in hematopoietic cells is downregulated by increasing temperature 2 degrees C. These observations provide a unique opportunity to identify cellular factor(s) which regulate BCR/ABL kinase in vivo and suggests possible novel treatment of CML by a mild hyperthermia. PMID- 10803517 TI - Immunotoxins against CD19 and CD22 are effective in killing precursor-B acute lymphoblastic leukemia cells in vitro. AB - Monoclonal antibodies (Mabs) conjugated to toxins or their subunits (immunotoxins or ITs) are undergoing clinical testing in adults with a variety of malignancies. The potential impact of this form of therapy in pediatric precursor B-lineage acute lymphoblastic leukemia (pre-B ALL) has yet to be determined. Mabs directed against the cell surface antigens, CD19 and CD22 conjugated to deglycosylated ricin A chain (dgRTA) have been tested in patients with non-Hodgkin's lymphoma (NHL), but not in patients with pre-B ALL. Because of the encouraging performance of these ITs in phase I trials, we evaluated the specific cytotoxicity of anti CD19 (HD37-dgRTA) and anti-CD22 (RFB4-dgRTA) ITs or their combination (Combotox) on patient-derived pre-B ALL cells maintained in vitro on a stromal feeder layer. After 48 h in culture, cytotoxicity to tumor cells was determined by flow cytometry using propidium iodide (PI) and fluorescein isothiocyanate (FITC) conjugated anti-CD10, 19, and 22. Both RFB4-dgRTA and HD37-dgRTA induced a statistically significant reduction in the number of viable leukemic cells, and Combotox was even more effective. Our results demonstrate that these ITs are specifically cytotoxic to primary pre-B ALL cells and that they should be further evaluated for the therapy of B-lineage ALL. PMID- 10803518 TI - HLA-DR4 is associated with a diminished risk of the development of chronic myeloid leukemia (CML). Chronic Leukemia Working Party of the European Blood and Marrow Transplant Registry. AB - CML is characterized by the chromosomal translocation t(9;22) (q34;q11) resulting in the chimeric bcr-abl oncogene that encodes P210 fusion proteins with novel amino acid sequences in the breakpoint region. If these peptides derived from P210 are presented by HLA molecules on the cell membrane of leukemic cells an immunological response may occur. Recent studies using synthetic peptides identical to the bcr-abl fusion region revealed that some peptides are capable of binding to the class I molecules HLA-A2,-A3,-A11 and -B8 and the class II molecules HLA-DR1, -DR2, -DR3, -DR4 and -DR11. Moreover T cell responses have been induced against bcr-abl-derived synthetic peptides bound to some of these HLA molecules. For HLA class I, we have previously shown that individuals expressing HLA-A3 and -B8 have a diminished risk of development of CML. To assess a similar protective effect of class II molecules we performed a large multi center study. This study compared the frequencies of HLA-DR1, -DR2, -DR3, -DR4 and -DR11 of patients with CML from the database of the EBMT (n = 1462) with unaffected individuals from the registry of Bone Marrow Donors Worldwide (n = 500 596). Patients and controls were matched per country. This analysis yielded significantly lower odds ratios (ORs) of 0.86 (95% CI 0.75-0.98) for HLA-DR3 and of 0.80 (95% CI 0.71-0.89) for HLA-DR4. The OR was 0.91 (95% CI 0.80-1.04) for HLA-DR1, 1.05 (95% CI 0.94-1.18) for HLA-DR2 and 0.87 (95% CI 0.74-1.01) for HLA DR11. To assess a possible effect of the linkage disequilibrium between HLA-B8 and HLA-DR3 we found that coexpression of HLA-B8 and HLA-DR3 gave an OR of 0.84 (95% CI 0.72-0.98), whereas HLA-DR3 positive/HLA-B8 negative individuals showed an OR of 1.02 (95% CI 0.84-1.24). This means that the protective effect of HLA DR3 of the development of CML was probably caused by its linkage disequilibrium with HLA-B8. In contrast, as there is no linkage disequilibrium of HLA-DR4 with HLA-A3 or HLA-B8, the results indicate that HLA-DR4 expression itself is associated with a diminished incidence of CML possibly by the presentation of bcr abl breakpoint peptides in these HLA molecules on the membrane of the leukemic cells. PMID- 10803519 TI - CDP/Cut DNA binding activity is down-modulated in granulocytes, macrophages and erythrocytes but remains elevated in differentiating megakaryocytes. AB - DNA binding by the CCAAT-displacement protein, the mammalian homologue of the Drosophila melanogaster Cut protein, was previously found to increase sharply in S phase, suggesting a role for CDP/Cut in cell cycle progression. Genetic studies in Drosophila indicated that cut plays an important role in cell-type specification in several tissues. In the present study, we have investigated CDP/Cut expression and activity in a panel of multipotent hematopoietic cell lines that can be induced to differentiate in vitro into distinct cell types. While CDP/Cut DNA binding activity declined in the pathways leading to macrophages, granulocytes and erythrocytes, it remained elevated in megakaryocytes. CDP/Cut was also highly expressed in primary megakaryocytes isolated from mouse, and some DNA binding activity could be detected. Altogether, these results raise the possibility that CDP/Cut may be a determinant of cell type identity downstream of the myelo-erythroid precursor cell. Another possibility, which does not exclude a role in lineage identity, is that CDP/Cut activity in megakaryocytes is linked to endomitosis. Indeed, elevated CDP/Cut activity in differentiating megakaryocytes and during the S phase of the cell cycle suggests that it may be required for DNA replication. PMID- 10803520 TI - All-trans-retinoic acid effects the growth, differentiation and apoptosis of normal human myeloid progenitors derived from purified CD34+ bone marrow cells. AB - We have previously shown that all-trans retinoic acid (ATRA) increases the number of CFU-GM colonies grown from unseparated human bone marrow cells with crude sources of colony stimulating factors. In this study, we further characterized the effect of ATRA on the growth of CFU-GM stimulated by individual cytokines from multiple samples of CD34+ enriched or purified human bone marrow cells. The number of IL-3- or GM-CSF-induced CFU-GM with 3 x 10(-7) M ATRA was 3.25+/-1.13, and 2.17+/-0.8-fold greater respectively, compared to controls without ATRA, while G-CSF had no effect and the ratio of colony-induced with or without ATRA was 1.06+/-0.17 (P = 0.00012). No colonies grew with ATRA + IL-6 or ATRA without a cytokine. Maximum enhancing effect on IL-3-induced CFU-GM occurred when ATRA was added on day 2, gradually diminished when delaying ATRA, and in cultures of day 9 or older adding ATRA had no effect. In 14 days liquid cultures of purified CD34+ cells with IL-3, ATRA increased the number of myeloid differentiated cells to 91-95%, compared to 37-70% with IL-3 alone. In addition, the number of apoptotic cells using the annexin V method increased after 14 days from 5.1% with IL-3 to 17.1% with IL-3 + ATRA and by the TUNEL in situ method from 10-26% to 60 95%, respectively. This study demonstrates that ATRA consistently enhances the growth of myeloid progenitors from CD34+ cells. This effect is dependent on the stimulating cytokine, suggesting the myeloid cells responding to ATRA are the less mature CFU-GMs that are targets of IL-3 and GM-CSF and not the G-CSF responding mature progenitors. The growth stimulation by ATRA and IL-3 is also associated with granulocyte differentiation and increased apoptosis. These studies further suggest a potential role of pharmacological doses of ATRA on the development of normal human hematopoietic cells. PMID- 10803521 TI - Effects of the chemokine stromal cell-derived factor-1 on the migration and localization of precursor-B acute lymphoblastic leukemia cells within bone marrow stromal layers. AB - Acute lymphoblastic leukemia (ALL) blasts undergo migration into layers of bone marrow fibroblasts (BMF) in vitro, utilizing the beta1 integrins VLA-4 and VL-5 as adhesion molecules. However, it has been unclear as to whether this is a selective process mediated by specific chemoattractant molecules, or simply a reflection of the highly motile nature of early B cell precursors. We further characterized this process using a transwell culture system, in which the two chambers were separated by an 8 microm diameter microporous membrane, through which leukemic cells could move. When a BMF layer was grown on the upper surface of the membrane there was an 84.1% reduction in transmigration of the human pre-B ALL cell line NALM-6 into the lower chamber, compared to control membrane with no BMF layer. Localization of leukemic cells under the BMF layer was confirmed ultrastructurally, suggesting the possibility that the migration of leukemic cells was directed by a chemotactic agent secreted by BMF. The involvement of the chemokine stromal cell-derived factor-1 (SDF-1) in this process was next investigated. BMF were shown to express m-RNA for SDF-1. Addition of SDF-1 at 100 ng/ml into the lower chamber increased transmigration of NALM-6 across the membrane by 2.2-fold, and also induced a 1.4- to 6.1-fold increase in movement of NALM-6 through a BMF layer into the lower chamber. The receptor for SDF-1, CXCR4, was demonstrated by flow cytometry on all 10 cases of precursor-B ALL analyzed, as well as on NALM-6, KM-3 and REH lines. An inhibitory antibody to CXCR4 was able to block the migration of NALM-6 cells into BMF monolayers grown on plastic by 51%, and in nine cases of ALL by 8-40%, as well as partially inhibit transmigration of leukemic cells through BMF layers along an SDF-1 concentration gradient. These results confirm that precursor-B ALL cells selectively localize within bone marrow stroma in vitro, and that this process is partially due to the stromal chemokine SDF-1 binding to its receptor CXCR4 on leukemic cells. SDF-1 may be important in influencing the localization of precursor-B ALL cells in marrow microenvironmental inches which regulate their survival and proliferation. PMID- 10803522 TI - Identification of variables determining the engraftment potential of human acute myeloid leukemia in the immunodeficient NOD/SCID human chimera model. AB - Among a variety of immunodeficient mouse strains the non-obese diabetic (NOD)/LtSz scid/scid strain appears to be most useful in allowing the engraftment of human AML. However, the large variability in ability to engraft and the levels of engraftment reached have not been explained. To address these issues we have investigated the NOD/SCID repopulating ability of 27 newly diagnosed AML samples. Patients were selected for the absence of internal tandem duplications in the Flt3 gene as we previously reported this mutation to be associated with an enhanced engraftment potential in this model. We observed that secondary AML (n = 6) had a significantly increased level of engraftment when compared to primary AML (n = 21, median levels 73.3% for secondary AML vs 8.94% for primary AML, P = 0.01). Within the primary AML, a significantly higher engraftment was observed in the FAB class M0 than in FAB classes M2, M4 and M5. Within primary AML, samples of patients who failed to respond to the initial therapy gave rise to a higher level of engraftment than samples of patients who did respond to therapy. A similar observation of an increased engraftment correlating with a poorer patient prognosis could be made when applying cytogenetic risk stratification. However, within the primary AML the most important clinical parameter correlating with the level of engraftment appeared to be the patient's WBC count at diagnosis (P = 0.0000). Covariate analysis with the WBC count as a covariate could also fully explain the differences observed in the cytogenetic risk groups, or on the basis of the initial therapy response. Although large differences could be observed, the ability to engraft the NOD/SCID mice was not linked to either the autonomous or cytokine-induced proliferation in vitro. As the leukemic cobblestone area forming cell frequencies also revealed no correlation with repopulation in the NOD/SCID model, we consider it very likely that the level of engraftment reflects the in vivo proliferative ability of the AML samples assayed rather than the number of leukemia-initiating cells infused into the NOD/SCID mice. Phenotypic analysis based on the expression of CD33, CD34 and CD38 before and after passage in NOD/SCID showed that in 10 out of 16 samples investigated phenotypes were different. In summary, in addition to the Flt3 internal tandem duplications we have identified a series of clinical parameters that determine the NOD/SCID repopulating ability of AML samples, whilst our data strongly suggest that AML in NOD/SCID does not reflect the leukemic process in the patient. PMID- 10803523 TI - Frequent disruption of the RB1 pathway in diffuse large B cell lymphoma: prognostic significance of E2F-1 and p16INK4A. AB - In the present study, we analysed 34 de novo diffuse large B cell lymphoma (DLCL) from a population-based lymphoma registry for alterations of the RB1 pathway at the genetic (RB1 and CDK4) and protein (pRb, cyclin D1, cyclin D3, CDK4, and E2F 1) level. The results were correlated with the data from our previous studies of CDKN2A deletion and hypermethylation, other p53 pathway components, p27Kip1 expression, and proliferation, as well as with clinical outcome, including prognosis. We found aberrant pRb expression in four (12%) of 34 DLCLs. One of these had a point mutation in intron 3 10 bp downstream of exon 3 generating a novel splice signal. Seven tumours (21%) showed cyclin D3 overexpression, including all three thyroid lymphomas (P = 0.006). Cyclin D3 overexpression and p16INK4A/pRb aberrations were mutually exclusive, supporting an oncogenic role for cyclin D3 in DLCL. p16INK4A inactivation, cyclin D3 overexpression, or aberrant pRb expression was identified in 18 of 34 DLCLs (53%). Combining these results with our previous p53 pathway studies showed that 82% of the de novo DLCLs had alterations of these pathways, and that both pathways were altered in 13 cases (38%). Low E2F-1 expression was associated with treatment failure (P = 0.020), and multivariate analysis of overall survival identified both low E2F-1 expression (relative risk = 6.9; P = 0.0037) and p16INK4A inactivation (relative risk = 3.3; P = 0.0247) as independent prognostic markers. These data support a role of E2F-1 as tumour suppressor gene in lymphoma and strongly suggest that the RB1 and p53 pathways are important in the development of de novo DLCL. Furthermore, low E2F-1 expression and p16INK4A inactivation may serve as prognostic markers for patients with this type of lymphoma. PMID- 10803524 TI - BCL10 gene mutations rarely occur in lymphoid malignancies. AB - BCL10, a gene involved in apoptosis signalling, has recently been identified through the cloning of chromosomal breakpoints in extranodal (MALT-type) marginal zone lymphomas carrying the t(1;14)(p22;q32) translocation. BCL10 was also found mutated in these cases as well as in other types of lymphoid and solid tumors, suggesting that its inactivation may play an important pathogenetic role; however, this has been questioned by recent studies showing a lack of somatic mutations in human cancers. We report the mutation analysis of exons 1-3 of the BCL10 gene in DNAs from 228 cases of lymphoid malignancies (30 B cell chronic lymphocytic leukemias, 123 B and 45 T non-Hodgkin's lymphomas and 30 multiple myelomas). Somatic mutations were detected in four cases (approximately 2%): one small lymphocytic, one follicular and two diffuse large cell lymphomas. The mutations were all within exon 3 and have not been previously reported. Our data suggest that BCL10 mutations may play only a limited role in the pathogenesis of lymphoid neoplasms. PMID- 10803525 TI - Clonal diversification of primary BALB/c plasmacytomas harboring T(12;15) chromosomal translocations. AB - DNA sequence analysis of PCR amplified Igh/c-myc junction fragments of T(12;15) chromosome translocations and immunohistochemical determination of immunoglobulin isotype production were employed to study the clonal diversification of neoplastic translocated plasma cells that resided in peritoneal inflammatory granulomas of BALB/c mice harboring primary plasmacytomas. The diversity of plasma cells was found to take two major forms when the fine structure of the T(12;15) translocation was used as the clonotypic marker. First, mosaics of clones containing translocations that were apparently unrelated to each other were detected in nine out of 17 (53%) mice. Second, subclones derived from common T(12;15)+ progenitors by either secondary deletions in translocation breakpoint regions or aberrant isotype switching near translocation breaksites were found in five of 17 (29.5%) mice. When Ig expression was utilized as the clonotypic marker, clonal mosaics were shown to occur in all mice. This was demonstrated by the finding that the prevalent IgA- or IgG-producing plasmacytoma clone was invariably accompanied by smaller clones of IgG- or IgA-expressing neoplastic plasma cells, respectively. These results provided new insights into the clonal diversification at the terminal stage of plasmacytomagenesis. In addition, they suggested that BALB/c plasmacytomas may be uniquely useful for studying clonal diversity during B cell oncogenesis, since clonal evolution can be evaluated in a pool of tumor and tumor precursor cells that is clearly defined by the T(12;15) chromosomal translocation and the production of monoclonal immunoglobulin. PMID- 10803526 TI - A novel natural killer cell line (KHYG-1) from a patient with aggressive natural killer cell leukemia carrying a p53 point mutation. AB - We present the establishment of a natural killer (NK) leukemia cell line, designated KHYG-1, from the blood of a patient with aggressive NK leukemia, which both possessed the same p53 point mutation. The immunophenotype of the primary leukemia cells was CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16+, CD56+, CD57+ and HLA-DR+. A new cell line (KHYG-1) was established by culturing peripheral leukemia cells with 100 units of recombinant interleukin (IL)-2. The KHYG-1 cells showed LGL morphology with a large nucleus, coarse chromatin, conspicuous nucleoli, and abundant basophilic cytoplasm with many azurophilic granules. The immunophenotype of KHYG-1 cells was CD1-, CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16-, CD25-, CD33+, CD34-, CD56+, CD57-, CD122+, CD132+, and TdT-. Southern blot analysis of these cells revealed a normal germline configuration for the beta, delta, and gamma chains of the T cell receptor and the immunoglobulin heavy-chain genes. Moreover, the KHYG-1 cells displayed NK cell activity and IL-2-dependent proliferation in vitro, suggesting that they are of NK cell origin. Epstein-Barr virus (EBV) DNA was not detected in KHYG-1 cells by Southern blot analysis with a terminal repeat probe from an EBV genome. A point mutation in exon 7 of the p53 gene was detected in the KHYG-1 cells by PCR/SSCP analysis, and direct sequencing revealed the conversion of C to T at nucleotide 877 in codon 248. The primary leukemia cells also carried the same point mutation. Although the precise role of the p53 point mutation in leukemogenesis remains to be clarified, the establishment of an NK leukemia cell line with a p53 point mutation could be valuable in the study of leukemogenesis. PMID- 10803527 TI - Immune and hematopoietic reconstitution after transplantation of cord blood progenitor cells: case report and review of the literature. AB - Transplantation using umbilical cord progenitor cells as the source of the stem cells is increasingly recognized as another form of allogeneic transplantation with curative intent. However, the different patterns of hematopoietic and immunological reconstruction have been described in very few patients. A 20-month old boy presented with acute leukemia. He received standard AML induction and consolidation therapy, after which he underwent allogeneic transplantation using HLA-matched sibling stem cells obtained from the umbilical cord. The preparative regimen consisted of busulfan and cyclophosphamide. White cell recovery, despite concomitant use of G-CSF, was slow, reminiscent of the engraftment pattern without the use of growth factor. Erythroid recovery was best recorded using fetal cell HbF level. Platelet transfusion independence occurred on day +31. Immunologic reconstitution revealed an early NK cell recovery by 6 weeks and progressive T cell recovery until 3 months, with continued increase in counts thereafter. However, the CD4/CD8 ratio remained low even at 14 months post transplantation. Recovery of B cells was slower until day +120. Proliferative response was within normal range on day +120. This report describes the unique engraftment pattern following umbilical cord blood transplant and emphasizes the pattern of immunological and hematological reconstitution. PMID- 10803528 TI - Detection of methylthioadenosine phosphorylase (MTAP) and p16 gene deletion in T cell acute lymphoblastic leukemia by real-time quantitative PCR assay. AB - Methylthioadenosine phosphorylase (MTAP) deficiency in tumors can be therapeutically exploited for selective therapy. Many tumors lacking MTAP have been found to homozygously delete the chromosome 9p region containing the p16 tumor suppressor gene. Several methods have been used to detect chromosome 9p deletions in primary tumors. However, the accurate diagnosis of chromosome 9p deletions has been hampered by the presence of contaminating normal cells. In search of an accurate and sensitive diagnostic method, we have developed the real time polymerase chain reaction assay using the TaqMan chemistry for quantitative detection of MTAP and p16 gene deletions. The assay's feasibility was tested with peripheral blood leukocytes (PBL) from 29 patients with adult T cell leukemia (ATL) previously analyzed with Southern blot analysis and validated on 39 PBL or bone marrow samples from childhood T cell acute lymphoblastic leukemia (T-ALL). Homozygous deletions of MTAP and p16 genes were detected respectively in six (20.7%) and eight (27.6%) of 29 ATL samples and in 15 (38.5%) and 23 (59%) of 39 T-ALL samples. The results correlated well with those of Southern blot analysis. It is of significance that the newly developed method can successfully detect homozygous deletions of these genes in samples containing as low as 33% blast cells. This rapid and sensitive method may be useful in searching for candidates for selective therapy targeting MTAP deficiency. PMID- 10803529 TI - Arsenic trioxide (As2O3) gradually downregulates tissue factor expression without affecting thrombomodulin expression in acute promyelocytic leukemia cells. PMID- 10803530 TI - Cellular cytotoxic drug sensitivity in children with acute leukemia and Down's syndrome: an explanation to differences in clinical outcome? PMID- 10803531 TI - Reported cryptic rearrangements of CBFbeta-MYH11 in acute myeloid leukaemia are not due to false priming of contaminating DNA. PMID- 10803532 TI - Tandem duplication of the FLT3 gene is infrequent in infant acute leukemia. Japan Infant Leukemia Study Group. PMID- 10803533 TI - Loss of CD38 antigen on CD34+CD38+ cells during short-term culture. PMID- 10803534 TI - Clonal chromosomal abnormalities identified by interphase fluorescence in situ hybridization on collagen cultured hematopoietic stem cells. PMID- 10803535 TI - Identification of PML/RARalpha rearrangements in suspected acute promyelocytic leukemia using fluorescence in situ hybridization of bone marrow smears: a comparison with cytogenetics and RT-PCR in MRC ATRA trial patients. MRC Adult Leukaemia Working Party. PMID- 10803536 TI - Low-dose melphalan in elderly acute myeloid leukaemia: complete remissions but resistant relapse with therapy-related karyotypes. PMID- 10803537 TI - Autografting with non-clonal mobilized hematopoietic progenitor cells in CML. PMID- 10803538 TI - Effects of sediment geochemical properties on the toxicity of copper-spiked sediments to the marine amphipod Gammarus locusta. AB - The contribution of some sediment features, namely acid-volatile sulfide (AVS) and total organic carbon (TOC) in order to explain the toxicity of copper-spiked sediments was investigated. A 10-day assay with the marine amphipod Gammarus locusta was performed with three selected sediment types that consisted of different proportions of fine fractions (FF) and TOC (0.5% FF/1% TOC; 25% FF/2% TOC; and 75% FF/4% TOC). Copper toxic effects were observed for 0.5% FF/25% FF sediments when simultaneously extracted copper (SEM(Cu)) minus acid volatile sulfide (SEM(Cu) - AVS) were 0.3 and 3.4 micromol/g, respectively. However, no significant mortality was observed for 75% FF sediment even when the sulfide binding capacity was exceeded (1.1 micromol/g < or = SEM(Cu) - AVS < or = 8.9 micromol/g). These results indicate that mortality could not be described solely as a function of the SEM(Cu) - AVS, suggesting the presence of other binding phases in addition to AVS. Results of the normalized LC50 values to total organic carbon indicated that this component probably also interferes in the amphipod survival, although not completely. The estimated LC50 values were close (2 mg Cu/g C and 8 mg Cu/g C, respectively for 0% FF and 25% FF sediment) although there was still a fourfold difference between them. Considering that none of these sediment features by itself could fully explain copper toxicity, an attempt was made to describe G. locusta mortality as a function of simultaneous SEM(Cu) - AVS and copper concentrations normalized to TOC. Nevertheless, the limited set of data available from this study prevented an attempt to model toxicity as a function of both of these factors. These findings reinforce the need to develop integrated models that include more than one binding phase, sulfide and organic carbon, and also other compartments of the system (e.g. interstitial water) to improve current methodologies for predicting copper toxicity based on sediment geochemical features. PMID- 10803539 TI - A system dynamic model for the assessment of different exposure routes in aquatic ecosystems. AB - With the exception of a limited number of highly persistent lipophilic pollutants, waterborne exposure is considered the most relevant exposure route for aquatic organisms, and even the only relevant exposure route in lower tier assessment, while the potential for bioaccumulation is only evaluated as secondary poisoning for birds and mammals. In addition, some evaluations consider that only the dissolved fraction is bioavailable for pelagic organisms. The lack of easy and cost-effective assessment methods is probably the main reason to disregard the contribution of other routes. This paper presents a system dynamics model that estimates the time-dependent accumulation of toxic chemicals through the food chain, and has also the potential to include the exposure from sediment. The generic ecosystem includes a primary producer and three levels of consumers, represented by unicellular algae, cladoceran invertebrates, cladoceran-eating fish and fish-eating fish, respectively. Nevertheless, the model, built using Microsoft Excel software, allows any number of levels and ecological-exposure relationships. The results obtained for four illustrative pollutants demonstrate that factors such as the binding to algae and other edible particles or food chain exposure for lipophilic non-persistent chemicals may constitute significant exposure routes. The current low tier European ecological risk assessment guidelines do not cover these aspects, and therefore may under estimate the real risk for both aquatic organisms and their predators. The model includes a simplified worst-case alternative that normally can be calculated using the existing information. Under a tiered approach, the simplified alternative is proposed to trigger the need of the toxicokinetics assays required to perform the complete time-dependent calculation. PMID- 10803540 TI - Potential use of flow cytometry in toxicity studies with microalgae. AB - Cytotoxic effects of aquatic pollutants on microalgae are very heterogeneous, and they are influenced by environmental conditions and the test species. Stress produced by copper or paraquat addition to the culture medium of two microalgae was analysed by flow cytometry. Parameters assayed were: cell volume, chlorophyll a fluorescence and cell viability. The variety of results obtained in the present study reveals that flow cytometry is a useful tool in the toxicity tests with microalgae, both marine and freshwater species, and for different kind of pollutants. PMID- 10803541 TI - In vitro response of the brown bullhead catfish (BB) and rainbow trout (RTG-2) cell lines to benzo[a]pyrene. AB - Established cell lines from rainbow trout (RTG-2) and brown bullhead catfish (BB) were evaluated as bioindicators of benzo[a]pyrene (B[a]P) toxicity with 3-(4,5 dimethyltiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction and neutral red (NR) uptake assays. Cytochrome P450 1A1 (CYP1A1) enzymatic activity was also evaluated, and taken as a biological indicator of the B[a]P induction power by ethoxyresorufin O-deethylase (EROD) and ethoxycoumarin O-deethylase (ECOD) assays. The BB and RTG-2 cells were compared after 1 and 6 days of exposure to B[a]P. The photoactivation of the compound (B[a]PUV) was another parameter taken into consideration. Cytotoxicity was not observed after 1 day of incubation with B[a]P in both cell lines, although the enzymatic activities of ECOD and EROD presented an induction. Apparently, after 1 day, cells did not metabolise sufficient amounts of B[a]P to cytotoxic metabolites. After 6 days of exposure to this compound a significant reduction in cell viability was observed, this reduction being superior to 50% at the highest B[a]P concentrations for the RTG-2 cell line. These results are in agreement with the values observed for the ECOD and EROD induction. The B[a]P cytotoxicity determined in both cell lines could be ascribed to the significant increase of EROD activity by 6 days of exposure. The photoactivation of B[a]P showed marked differences in both cytotoxic assays and CYP1A1 enzymatic activities, for both cell lines. After 1 day of exposure there was a significant reduction in cell viability, superior to 50% for the RTG-2 cell line. However, it was observed that no induction occurred but rather a decrease in ECOD and EROD activities. Six days of incubation with B[a]PUV showed a decrease in cell viability at the highest concentrations for the BB cells and at the lowest concentrations for the RTG-2 cell line, and the CYP1A1 enzymatic activity presented a significant induction. These results and those observed after 1 day of exposure suggest that B[a]PUV acts as a direct-acting toxicant as well as a metabolism-mediated toxicant-like B[a]P. The RTG-2 cells were more sensitive to B[a]P and its toxic metabolites as well as to the photoactivation of the compound, in both exposure times tested. The finding that the cell lines responded to the CYP1A1 induction in a very efficient way gives proof of the applicability of this system to environmental biomonitoring and toxicology. PMID- 10803542 TI - In vitro and in vivo inhibition of Daphnia magna acetylcholinesterase by surfactant agents: possible implications for contamination biomonitoring. AB - This study was designed to investigate the effect of two surfactants, dodecyl benzyl sulfonate (DBS), sodium dodecyl sulfate (SDS), and of a domestic detergent (Y) on the AChE activity of the crustacean cladoceran Daphnia magna. All the chemicals significantly inhibit the activity of the enzyme, both in vitro and in vivo conditions. In vitro lowest observed effect concentration (LOEC) values ranged from 12.5 to 100 mg/l and correspondent IC50 (50% inhibition concentration) values ranged from 6.6 to 58.5 mg/l. In vivo LOEC values ranged from 2 to 11.9 mg/l, while EC50 (50% effect concentration) values ranged from 11.4 to 56.7 mg/l. AChE inhibition by environmental contaminants such as surfactants, detergents and metals may lead to false diagnostics and even wrong conclusions in biomonitoring studies based on the use of AChE as a specific biomarker for organophosphorous and carbamate compounds. PMID- 10803543 TI - Rapid and cost-effective multiparameter toxicity tests for soil microorganisms. AB - Three biochemical parameters, DNA quantification in soil samples and two enzymatic activities, beta-galactosidase and dehydrogenase have been assessed as potential end-points for the use in cost-effective toxicity tests on soil microorganisms. The assessment included the development of a classical dose response 24-h assay and the incorporation of measurements of the effects on microbial activities in soil column leaching studies and multispecies miniaturised terrestrial systems (MTS). Four different chemicals, copper, a new herbicide, thiabendazole and fenthion were studied. A rapid fluorescence DNA quantification technique did not produce adequate responses. The efforts to quantify DNA after extraction and clean-up procedures failed due to the presence of humic acids. From the protocol of the technique one could see that the technical procedure is time-consuming and expensive and, for this reason, not suitable for use as a parameter in rapid and cost-effective tests. However, the enzymatic activities showed their potential as toxicity end-points. Copper produced a concentration/response inhibition of beta-galactosidase and dehydrogenase with EC50 values of 78.39 and 24.77 mg Cu/kg soil, respectively. In the soil column study, these endpoints allowed the measurement of the microbial activities through the column. The effects of the new herbicide on beta galactosidase and dehydrogenase activities were statistically significant for the highest application dose (40 g/ha). Thiabendazole affected the microbial activity when mixed within the soil, but no effects were observed when this fungicide was applied on the soil surface. Fenthion produced effects when applied either in the soil or on the soil surface. These results can be explained by the low mobility of thiabendazole. The results show the capabilities of these biochemical parameters to be included as endpoints in cost-effective bioassays. PMID- 10803544 TI - Assessing the potential hazard of chemical substances for the terrestrial environment. Development of hazard classification criteria and quantitative environmental indicators. AB - Hazard assessment constitutes an essential tool in order to evaluate the potential effects of chemical substances on organisms and ecosystems. It includes as a first step, hazard identification, which must detect the potential dangers of the substance (i.e. the kind of effects that the substance may produce), and a second step to quantify each danger and to set the expected dose/response relationships. Hazard assessment plays a key role in the regulation of chemical substances, including pollution control and sustainable development. However, the aquatic environment has largely received more attention than terrestrial ecosystems. This paper presents the extrapolation of several basic concepts from the aquatic to the terrestrial compartment, and suggests possibilities for their regulatory use. Two specific proposals are discussed. The first focuses on the scientific basis of the hazard identification-classification criteria included in the EU regulations and their extrapolation to the terrestrial environment. The second focuses on the OECD programme for environmental indicators and the development of a soil pollution pressure indicator to quantify the potential hazards for the soil compartment and its associated terrestrial ecosystem related to the toxic chemicals applied deliberately (i.e. pesticides) or not (i.e. heavy metals in sludge-based fertilisers; industrial spills) to the soil. PMID- 10803545 TI - Beyond the Mussel Watch--new directions for monitoring marine pollution. AB - The increasing numbers and types of potential pollutants in the world oceans calls for novel strategies for their assays. The 'Mussel Watch' well served the latter decades of the 20th century. For the immediate future, individual assays of the chemicals of concern should be replaced by the analyses of groups of pollutants which have a common impact upon marine organisms. Secondly, more attention should be focussed upon the benthos where many potential pollutants continue to accumulate. Impacts upon members of the marine biosphere may be recognized by population changes of individual species. Lastly, time frame monitoring studies should be initiated, so that long-term trends in the health of a system can be detected. PMID- 10803546 TI - Changes in mussel biometry on exposure to metals: implications in estimation of metal bioavailability in 'Mussel-Watch' programmes. AB - The occurrence of changes in flesh and shell weights and in other biometric parameters of mussels, Mytilus galloprovincialis, has been related to different metal levels found in their soft tissues. The effects of clean and Zn-polluted environments and laboratory experiments where Zn-polluted mussels were exposed to sublethal concentrations of Zn, Cu and Cd were investigated. Zinc-polluted mussel shell weights increased significantly after a 51-day depuration period. Exposure of Zn-polluted mussels to Zn or Cd, however, caused a slightly reduced shell growth in comparison with depurating mussel Cu-exposures not causing any reduction in growth. Apart from metal concentrations, metal/shell weight indices have been used to assess metal bioavailability. Metal concentrations recorded in the soft tissues of depurating mussels increased without a source of 'extra' metals, while the Zn/shell-weight index was reduced, as expected from a depuration process, Cu and Cd/shell-weight indices remaining constant. Experimental exposure to Zn, Cu and Cd caused augmented values of Zn, Cu and Cd/shell-weight indices, respectively. These different findings were attributed to changes in flesh weight (related with gamete spawning) that would produce inconsistent estimates of whole metal concentration in soft tissues. Since changes in the tissue composition and in growth rates do not affect Zn/Cu ratios and metal/shell-weight indices, these parameters are proposed as reliable indices of metal bioavailability for 'Mussel-Watch' monitoring programmes. The most sensitive parameter is the metal/shell-weight index, which is, therefore, highly recommended to be used in 'Mussel-Watch' monitoring programmes in order to determine metal bioavailability in seawaters. PMID- 10803547 TI - Heavy metal fluxes at the sediment-water interface of three coastal ecosystems from south-west of the Iberian Peninsula AB - Concentrations of the heavy metals Cr, Cu, Fe and Mn were measured in sediments and porewater samples collected in three coastal ecosystems southwest of the Iberian Peninsula: the Odiel and Barbate River Salt Marshes and the Bay of Cadiz. Both the sediment and the porewater metal concentrations in the Odiel River Salt Marshes are higher than the values found in the Bay of Cadiz and Barbate River Salt Marsh, particularly for copper, a metal associated with mining activity. In porewater, the profiles were not the same as those in the solid phase and reflect the different behaviours of the elements in relation to the redox conditions. The heavy metals Cr and Cu show a typical enrichment in the porewater of the oxic zone. The heavy metals Mn and Fe show an increase in the porewater at the depths where the maximum nitrate and phosphate concentrations occur, respectively. Significant differences between background levels for each heavy metal in the various studied zones exist. Iron and Cu showed larger background levels in the Odiel River Salt Marshes than those in the Cadiz Bay and the Barbate River Salt Marshes. In the Bay of Cadiz the background levels are also high, particularly for Cr. At the Odiel River Salt Marshes the diffusive flux of Cu is high (1.3 230.1 microg cm(-2) year(-1)), which suggests that the Odiel River Salt Marshes are subject to strong contamination by Cu, which is presumably introduced to the sediment in particulate form. In the Bay of Cadiz, Cr is the only metal with positive diffusive flux (2.15 microg cm(-2) year(-1)). It is higher than those obtained in other coastal ecosystems including the Odiel River Salt Marshes. The positive diffusive flux of Cr has been associated with the input of this metal by the naval industry and the manufacturing of car and aircraft components. PMID- 10803548 TI - Use of polyclonal antibodies for the detection of changes induced by cadmium in lysosomes of aquatic organisms. AB - Lysosomal responses are widely accepted cellular effect biomarkers of general stress. Up to now, these biomarkers have been analysed by means of conventional techniques based on enzyme histochemical methods, where lysosomal enzymes such as acid phosphatase and beta-glucuronidase (beta-GUS) have been employed as markers of lysosomes. The aim of the present work was to develop more advanced and sensitive methods based on the use of polyclonal antibodies to measure lysosomal enzymes in different sentinel organisms. For this purpose, we have studied the cross-reactivity of two commercial polyclonal antibodies against the lysosomal enzymes acid phosphatase and beta-GUS with molluscan digestive gland by means of immunoblotting and immunohistochemistry. The antibody against acid phosphatase cross-reacted specifically with the lysosomal fraction of the digestive gland, while unspecific immunoreaction occurred with digestive gland whole homogenates and tissue sections. The antibody against beta-GUS cross-reacted specifically with digestive gland whole homogenates and tissue sections. The cross-reactivity of this antibody was tested also in crab hepatopancreas and mullet liver where the same successful results were obtained. The second aim of the present study was to test if the immuno-based approach was sensitive enough to detect lysosomal alterations provoked by contaminants. For this purpose two experiments were carried out with mussels treated with cadmium in two ways: in vivo treatment by injection and in vitro treatment using digestive gland explants. Afterwards immunoblotting studies with the antibody against beta-GUS were applied and immunoreactive bands were quantified by means of a gel analysis programme. We found that beta-GUS protein levels were higher in treated mussels when compared with controls in either in vivo or in vitro treatments. All these data suggest that the polyclonal antibody against beta-GUS is adequate to be used in immuno based approaches to detect contaminant-induced lysosomal alterations. PMID- 10803549 TI - New methods to use fish cytochrome P4501A to assess marine organic pollutants. AB - A new methodology has been developed to assess cytochrome P4501A expression in two South Atlantic Spanish fish, guilthead seabream (Sparus aurata) and grey mullet (Liza aurata), used as pollution bioindicators. Degenerate oligos were used to amplify by reverse transcription and PCR (RT-PCR) specific cyp1A cDNA sequences, used subsequently to design specific primers to get the full cDNA by rapid amplification of cDNA ends. A new assay has been developed to quantitate cyp1A expression by RT-PCR in an automated DNA sequencer. The effect of beta naphthoflavone inducing biotransformation has been used to compare three distinct pollution biomarkers: EROD activity, ELISA determination of CYP1A, and 2 aminoanthracene (2-AA) activation. Immunodetection by ELISA or Western blot was inconsistent in S. aurata and L. aurata. EROD activity yielded satisfactory results; the higher induction was observed by bioactivation of 2-AA to mutagens detected with strain BA149 of Salmonella typhimurium, in agreement with the high sensitivity previously described for this biomarker. The present paper summarizes the current status of our research. PMID- 10803550 TI - The use of Nucella lapillus (L.) transplanted in cages to monitor tributyltin (TBT) pollution. AB - Specimens of the gastropod Nucella lapillus were transplanted from their natural habitat to sites affected by tributyltin (TBT) pollution to diverse degrees; individuals were held in cages that also included mussels as food. Subsamples were taken from the cages and the natural site at different time intervals (1, 2 and 5 months) to compare growth, imposex (a set of sexual anomalies caused by TBT) and butyltin concentrations accumulated in female tissues. Individuals in cages experienced negligible mortality and considerable growth. In addition, while the Relative Penis Size Index (RPSI, an index of imposex) remained consistent in the natural site through the experiment, it increased markedly at the implant sites; the same applies to the butyltin residues. These results are discussed to conclude that cage transplanting of N. lapillus is a suitable technique that can quickly (i.e. 1 month) detect differences in the level of TBT contamination among sites; however, extrapolation of experimental results to the field seems to require a longer exposure time (at least 5 months). Finally, the use of such an approach is deemed helpful to interpret the relationship between N. lapillus population dynamics and TBT pollution. PMID- 10803552 TI - Biotic indices based on soil nematode communities for assessing soil quality in terrestrial ecosystems. AB - Nematode communities offer several advantages for assessing the quality of soils. Diversity and maturity indices as well as trophic structure defining parameters based on the composition of soil nematode communities have been used to detect various kinds of man-induced disturbances and monitor ecological recovery. In the present paper we examined the response of these indices in herbaceous Mediterranean ecosystems. Incipient soils, which were developed on the top of waste dumping sites were compared with agricultural soils under fallow. Grasslands and old-fields were considered as reference systems. Either diversity or maturity indices were shown to be very sensitive to changes in soil system caused by waste dumping. However, they were less useful for the categorisation of perturbation in agroecosystems. Trophic composition related parameters were more meaningful for this aim. Plant parasite index (PPI) may be a good generic indicator of recovery processes after perturbation in these annual herbaceous ecosystems. PMID- 10803551 TI - The use of two species of crayfish as environmental quality sentinels: the relationship between heavy metal content, cell and tissue biomarkers and physico chemical characteristics of the environment. AB - In systems as heavily disturbed as rivers, the use of biological sentinels is a most interesting way of obtaining continuous assessment of environmental quality. This study seeks to establish the value of such sentinels of two species of crayfish: the native crayfish (Austropotamobius pallipes, Lereboullet 1858) and an introduced species, signal crayfish (Pacifastacus leniusculus, Dana 1852), by comparing the habitat quality (water and sediments) and heavy metal contents in crayfish with the degree of stress manifested by the animals, measured by cell and tissue biomarkers. For this purpose a histological study of the digestive gland was carried out together with measurements of heavy metal contents in samples from various points of the river network of Bizkaia (Spain), selected on the basis of their degree of disturbance. The results establish a positive correlation between these environmental conditions, the cell and tissue biomarkers of the digestive gland (thinning of the digestive epithelium, enlargement of digestive lysosomes) and the heavy metal contents of the animals (converted to an index of pollutant load). PMID- 10803553 TI - Histopathology of the digestive gland of an Antarctic limpet exposed to cadmium. AB - Histopathological alterations induced experimentally with cadmium (Cd) in Antarctic limpets (Nacella concinna), exposed for different times and concentrations were compared to controls. At the light microscope level, samples exposed to the contaminant for short periods (6, 12 and 24 h) at two different concentrations (0.25 and 0.5 mg l(-1)) showed no alterations compared to controls. After 48 h of exposure at a 0.5 mg l(-1) Cd concentration, vacuolisation of the basophilic cells was observed. After 72 h exposure, there was a marked loss of all the digestive gland structure, with cell autolysis and loss of basophilia. PMID- 10803554 TI - Ecological risk assessment of pesticides in the Mediterranean region. The need for crop-specific scenarios AB - The risk assessment procedures of pesticides in the European Union are mostly based on worst-case scenarios developed for central and northern European conditions. From the point of view of Mediterranean conditions, these assessment procedures are not always appropriate, and therefore the development of specific scenarios is required. There are differences not only in meteorological data (temperature, solar irradiation, or rainfall volume and annual distribution) between these countries, but also in farm distribution, crop characteristics, soil or sediment properties and surface or groundwater characteristics. These differences are more evident for the typical crops of the southern countries, such as olive groves, vineyards, citrus or rice fields, and in these cases more realistic estimates of exposure are required. Our proposal includes the development of new criteria to improve the ecological risk assessment process for the conditions of the Mediterranean region, and the justification of this need. PMID- 10803555 TI - Deriving ecological risk-based soil quality values in the Basque Country. AB - Most industrialised countries are currently drawing up their procedures to deal with contaminated land. In the Basque Country generic soil quality values applicable to different land uses have been drawn up for many common soil inorganic and organic pollutants, to assist in the site assessment process. In this paper we present those values aimed at protecting ecosystems as the main target at risk. Ecological risk was defined on the basis of effects on the structure of the soil invertebrate community. Ecotoxicological data from laboratory tests were used in combination with a theoretical extrapolation method. The following groups of compounds were assayed: heavy metals, cyanides, aromatic compounds, polycyclic aromatic hydrocarbons, HCH isomers, chlorinated organic compounds, and mineral oil. A specific toxicity database was created. The reliability of the values obtained as well as the limitations of the derivation process are discussed. PMID- 10803556 TI - Organosmectites as sorbent and carrier of the herbicide bentazone AB - Sorption isotherms of bentazone on diverse organoclays (OCls) were measured at aqueous concentrations from 25 to 1500 microM. Sorption on organophilic OCls, OCls with large quaternary alkylammonium groups, i.e. hexadecyltrimethylammonium (AHDT) and dioctadecyldimethylammonium (ADOD), were compared to that on sorptive OCls, which have large primary alkylammonium (octadecylammonium, AC18) and small quaternary alkylammonium (phenyltrimethylammonium, PTA) groups. The organophilic OCls showed much higher sorption (AHDT Kd = 682-1789; ADOD Kd = 838-1728) than the sorptive OCls (AC18 Kd = 38-40; PTA Kd = 0), suggesting hydrophobic bentazone organocation interactions as the main mechanism responsible for sorption. The incubation of the two organophilic OCls with soil contaminated with bentazone showed that availability of the herbicide decreased almost instantaneously from 124 microg g(-1) to 1 microg g(-1). Bentazone sorbed on the OCl, and thus immobilized, was partially extractable with CaCl2/methanol solution suggesting its potential bioavailability. Two OCls, with high and low sorptions, were assayed as herbicide carriers by preparing bentazone-OCl complexes and monitoring their herbicide release in water and soil-water suspensions. These bentazone-OCl complexes released 20-80% of their bentazone content, depending of the sorbent capacity and the bentazone-OCl interaction. Results of this work show that by varying the sorptive properties of OCls, they can be used to immobilize pesticide in a contaminated soil and to protect soil and water by using OCl as pesticide carriers in slow release formulations. PMID- 10803557 TI - The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian Peninsula: a practical approach. AB - Within the frame of the 2nd Iberian Congress of Environmental Contamination and Toxicology (University of the Basque Country, Leioa, June 1998) a workshop was held about the use of biomarkers in marine pollution monitoring. Among others, the following biomarkers received special attention: metallothionein induction, acetylcholinesterase inhibition, cytochrome P450 system induction, imposex, lysosomal enlargement and lysosomal membrane destabilisation, and peroxisome proliferation. These biomarkers can be used to evaluate exposure to and effect of different contaminants (metals, organic xenobiotics and organometallic compounds) and they can be measured using different methodological approaches (biochemistry, cytochemistry, immunochemical methods based on the use of biotechnology). Before the application of a set of biomarkers in pollution monitoring programmes, well defined protocols of Quality Assurance have to be established to allow adequate comparison of results. It is also necessary to include analysis of standard reference materials and to obtain detailed knowledge of basal values and seasonal variations of the biomarkers in various species, as well as to integrate the information obtained with the different biomarkers. Marine bivalve molluscs such as mussels are appropriate sentinel species for most of the biomarkers proposed except for the induction of the cytochrome P450 system, which should be measured in fish, and the degree of imposex, which is a biomarker of exposure to TBT specifically measured in certain gastropod molluscs. As a result of the workshop, a battery of biomarkers of contaminant exposure and effects are proposed that could be incorporated into programmes monitoring the quality of the coastal environment in the Iberian Peninsula. These measures would be undertaken in conjunction with chemical measures of contaminant burdens in selected sentinel species. PMID- 10803558 TI - Cytochrome P4501A (CYP1A) in teleostean fishes. A review of immunohistochemical studies. AB - Cytochrome P4501A monooxygenase has an important function in the biotransformation of many xenobiotics, including polynuclear aromatic hydrocarbons, and planar organochlorine compounds. The metabolism can lead to detoxification or activation to reactive intermediates. Exposure of fish leads to a receptor-mediated induction of CYP1A gene expression. The induction response can be quantitatively analysed by means of molecular techniques (RT-PCR, Northern Blotting), immunochemical approaches (ELISA, Western Blotting), and enzymatic methods (7-ethoxyresorufin-O-deethylase, EROD) at the catalytical level. Immunohistochemical studies have provided qualitative information on cell and tissue distribution of CYP1A in teleost fish. The liver is the major organ of CYP1A activity in fish, but the enzyme is additionally expressed in numerous extrahepatic organs, including kidney, alimentary canal, heart, gills, olfactory system, gonads, brain and endocrine tissues. In many tissues, the vascular endothelia show a strong CYP1A immunoreactivity. As indicated from immunohistochemical studies with fish embryos and larvae, the typical cell and tissue distribution of CYP1A is established early during fish ontogeny. PMID- 10803559 TI - Evolution and coordination of the alternative methods in Spain AB - The present work is the introductory contribution to the workshop on 'Alternative methods for toxicity assessment' held during the 2nd Iberian Congress of Environmental Contamination and Toxicology (University of the Basque Country, Leioa, June 1998). Other contributions presented at the workshop are reported by Castano et al., Repetto et al. and Prieto in this volume. In this introductory contribution we have highlighted the different initiatives brought about in Spain to coordinate the studies about alternative methods for toxicity assessment. The first initiative was the meeting of the ICLAS/CSIC Working Group on Complementary Methods, held in Talavera de la Reina in 1995. A list of validated methods was published in a monograph as a result of the meeting. Recently a Spanish net has been established for the development of alternative methods (Spanish Network for the Development of Alternative Methods or Red Espanola para el Desarrollo de Metodos Alternativos, REMA). The first meeting for the Constitution of REMA will be held in December 1999 in Madrid. The REMA, as organising entity, consists of scientific and industrial societies or industrial associations, as well as observers from the Administration, and encourages the participation of all those interested on an individual basis. PMID- 10803560 TI - The use of alternative systems for the ecotoxicological screening of complex mixtures on fish populations. AB - This paper presents the results of the use of alternative systems in a screening study of four complex mixtures. The following tests were performed: in vitro induction of micronuclei in a rainbow trout-derived cell line by flow cytometry, and hatching percentage, time of hatching and teratogenic alterations on the embryolarval development on medaka fish eggs. The results obtained with the proposed tests in this study allows an increase in the information level in a short period of time (2 weeks), using very low sample volumes (< 100 ml). Inclusion of chronic and specific effects (genotoxicity and teratogenicity) allows the selection of the most sensitive endpoint to increase security factors in the ecotoxicological assessment of complex mixtures, so that detailed studies can be focused only on those samples which require further research. PMID- 10803561 TI - ECVAM's in-house prevalidation/validation studies in the areas of haematotoxicity, reproductive toxicity, metabolism-mediated toxicity and epithelial barrier function. AB - The European Centre for the Validation of Alternative Methods (ECVAM) facilitates, co-ordinates and participates in validation activities at the European Union level. Various experimental studies, e.g. in the areas of haematotoxicity, reproductive toxicity, nephrotoxicity and epithelial barrier function, and metabolism-mediated toxicity, are underway in ECVAM's laboratories. ECVAM itself is currently involved in the prevalidation/validation of two assays, the colony-forming unit granulocyte/macrophage (CFU-GM) assays for predicting acute neutropenia and the embryonic stem cell test for predicting embryotoxicity. In the areas of metabolism-mediated toxicity and nephrotoxicity and epithelial barrier function, several assays are in the course of development. In many cases, the recommendations of various ECVAM workshops are being followed. PMID- 10803563 TI - OECD environmental data 1999 compendium PMID- 10803562 TI - A higher tier flow-through toxicity test with the green alga Selenastrum capricornutum. AB - A flow-through test system for the assessment of the toxicity of substances to the green alga Selenastrum capricornutum was developed. The dosing of a test substance commenced, once the growth of the algae was in a steady state, i.e. when the dilution rate was equal to the growth rate. The growth rate and hence the effect of the test substance was determined indirectly by the estimation of cell number under consideration of inflow volume over time. The ease of manipulation of the exposure regime via digitally controlled pumps makes this flow-through system also suitable for testing substances under realistic exposure conditions or in combination with their metabolites. In this study the degradation of a herbicide and its metabolites was simulated. The substance with an ErC50 (72 h) of 1.3 microg/l under static conditions (OECD, 1984) was tested in the developed flow-through system. The simulated degradation at an initial dose of 3 microg/l of the herbicide had no effect on algal growth, whereas under static test conditions, an NOErC of 0.28 microg/l was determined. The lowest concentration at which effects were observed was a continuous dose of 6 microg/l. PMID- 10803564 TI - Steroid transformation enzymes as critical regulators of steroid action in vivo. PMID- 10803565 TI - Moments in time--the neonatal rat hypothalamo-pituitary-adrenal axis. PMID- 10803566 TI - Rapid induction of corticotropin-releasing hormone gene transcription in the paraventricular nucleus of the developing rat. AB - Neonates from postnatal days (pnd) 4-14 display a minimal pituitary-adrenal response to mild stress, the so-called stress hyporesponsive period (SHRP). However, during the SHRP, maternal deprivation (deprived) alters the pituitary adrenal system, enabling neonates to become endocrine responsive to specific stimuli. Although neonates do display stress-induced ACTH, there is limited evidence for enhanced CRH gene expression early in development. The present experiment examined whether a mild stimulus (isotonic saline injection) administered to deprived and nondeprived neonates would enhance CRH biosynthesis in the paraventricular nucleus. Using in situ hybridization we measured the time course of CRH heteronuclear RNA (hnRNA) and messenger RNA at 15, 30, and 240 min poststimulus. Pnd 6, 12, and 18 were included to examine the CRH gene response during and outside of the SHRP. Despite the minimal endocrine response of nondeprived pups during the SHRP, CRH hnRNA and messenger RNA were elevated at 15 min (all ages). Both transcripts were enhanced at 15-30 min in deprived (pnd 12 and 18) pups; however, the magnitude of the response was less than that in nondeprived pups. These data indicate that during ontogeny there is a rapid stimulus-induced CRH biosynthesis. Thus, during development, the central components of the hypothalamic-pituitary-adrenal axis may be stress hyperresponsive rather than hyporesponsive. PMID- 10803567 TI - Regulation by adrenocorticotropin (ACTH), angiotensin II, transforming growth factor-beta, and insulin-like growth factor I of bovine adrenal cell steroidogenic capacity and expression of ACTH receptor, steroidogenic acute regulatory protein, cytochrome P450c17, and 3beta-hydroxysteroid dehydrogenase. AB - The purpose of this study was to evaluate the time-course effect of a 36-h treatment with ACTH (10(-8) M), transforming growth factor-beta1 (TGFbeta1; 10( 10) M), angiotensin II (AngII; 10 (-7) M), and insulin-like growth factor I (IGF I; 10(-8) M) on the steroidogenic capacity of bovine adrenocortical cells (BAC) and on messenger RNA (mRNA) levels of ACTH receptor, cytochrome P450c17, 3beta hydroxysteroid dehydrogenase (3betaHSD), steroidogenic acute regulatory protein (StAR), and StAR protein. ACTH and IGF-I enhanced, in a time-dependent manner, the acute 2-h ACTH-induced cortisol production, whereas TGFbeta 1 and AngII markedly reduced it. ACTH, IGF-I, and AngII increased ACTH receptor mRNA, but the opposite was observed after TGFbeta1 treatment. ACTH and IGF-I increased P450c17 and 3betaHSD mRNAs, whereas AngII and TGFbeta1 had the opposite effects. However, the effects of the four peptides on ACTH-induced cortisol production appeared before any significant alterations of the mRNA levels occurred. The most marked and rapid effect of the four peptides was on StAR mRNA. The stimulatory effect of ACTH was seen within 1.5 h, peaked at 4-6 h, and declined thereafter, but at the end of the 36-h pretreatment, the levels of StAR mRNA and protein were higher than those in control cells. IGF-I also enhanced StAR mRNA levels within 1.5 h, and these levels remained fairly constant. The effects of AngII on StAR mRNA expression were biphasic, with a peak within 1.5-3 h, followed by a rapid decline to almost undetectable levels of both mRNA and protein. TGFbeta1 had no significant effect during the first 3 h, but thereafter StAR mRNA declined, and at the end of the experiment the StAR mRNA and protein were almost undetectable. Similar results were observed when cells were treated with ACTH plus TGFbeta1. A 2-h acute ACTH stimulation at the end of the 36-h pretreatment caused a higher increase in StAR mRNA and protein in ACTH- or IGF-I-pretreated cells than in control cells, which, in turn, had higher levels than cells pretreated with TGFbeta1, ACTH plus TGFbeta1, or AngII. These results and the fact that the stimulatory (IGF-I) or inhibitory (AngII and TGFbeta1) effects on ACTH-induced cortisol production were more pronounced than those on the ability of cells to transform pregnenolone into cortisol strongly suggest that regulation of StAR expression is one of the main factors, but not the only one, involved in the positive (IGF-I) or negative (TGFbeta1 and AngII) regulation of BAC for ACTH steroidogenic responsiveness. A high correlation between steady state mRNA level and acute ACTH-induced cortisol production favors this conclusion. PMID- 10803568 TI - Identification of the oxidative 3alpha-hydroxysteroid dehydrogenase activity of rat Leydig cells as type II retinol dehydrogenase. AB - Dihydrotestosterone (DHT) is the most potent naturally occurring androgen, and its production in the testis may have important consequences in developmental and reproductive processes. In the rat testis, three factors can contribute to intracellular DHT levels: 1) synthesis of DHT from T by 5alpha-reductase, 2) conversion of DHT to 5alpha-androstane-3alpha, 17beta-diol (3alpha-DIOL) by the reductive activity of 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD), and 3) conversion of 3alpha-DIOL by an oxidative 3alpha-HSD activity. While the type I 3alpha-HSD enzyme (3alpha-HSD1 or AKR1C9) is an oxidoreductase in vitro and could theoretically be responsible for factors 2 and 3, we have shown previously that rat Leydig cells have two 3alpha-HSD activities: a cytosolic NADP(H)- dependent activity, characteristic of 3alpha-HSD1, and a microsomal NAD(H)-dependent activity. The two activities were separable by both developmental and biochemical criteria, but the identity of the second enzyme was unknown. To identify the microsomal NAD(H)-dependent 3alpha-HSD in rat Leydig cells, degenerate primers were used to amplify a number of short-chain alcohol dehydrogenases. Sequence analysis of cloned PCR products identified retinol dehydrogenase type II (RoDH2) as the prevalent species in purified Leydig cells. RoDH2 cDNA was subcloned into expression vectors and transiently transfected into CHOP and COS-1 cells. Its properties were compared with transiently transfected 3alpha-HSD1. When measured in intact CHOP and COS-1 cells, RoDH2 cDNA produced a protein that catalyzed the conversions of 3alpha-DIOL to DHT and androsterone to androstanedione, but not the reverse reactions. Therefore, the 3alpha-HSD activity of RoDH2 was exclusively oxidative. In contrast, type I 3alpha-HSD cDNA produced a protein that was exclusively a 3alpha-HSD reductase. In cell homogenates and subcellular fractions, RoDH2 catalyzed both 3alpha-HSD oxidation and reduction reactions that were NAD(H) dependent, and the enzyme activities were located in the microsomes. Type I 3alpha-HSD also catalyzed both oxidation and reduction, but was located in the cytosol and was NADP(H) dependent. We conclude that type I 3alpha-HSD and RoDH2 have distinct 3alpha-HSD activities with opposing catalytic directions, thereby controlling the rates of DHT production by Leydig cells. PMID- 10803569 TI - Thyroid hormone and estrogen regulate brain region-specific messenger ribonucleic acids encoding three gonadotropin-releasing hormone genes in sexually immature male fish, Oreochromis niloticus. AB - The present study was undertaken to determine whether T3, estrogen, and 11 ketotestosterone could alter a specific population of GnRH-containing neurons, as indicated by a change in messenger RNA (mRNA) levels in sexually immature male tilapia, Oreochromis niloticus. Two weeks after castration, fish were assigned to four treatment groups. One group served as the control (sesame oil); a single ip injection of (T3; 5 microg/g), estradiol benzoate (EB; 5 microg/g), or 11 ketotestosterone (KT; 5 microg/g) was administered to the remaining three groups. Twenty-four hours after the injection, brains were collected and processed for in situ hybridization histochemistry using 35S-labeled 30-mer antisense oligonucleotide probes complementary to the GnRH-coding region of chicken II, salmon, and seabream GnRH. Computerized image analysis was performed to quantify mRNA concentrations, neuronal numbers, and neuronal size of the terminal nerve nucleus olfactoretinalis, preoptic, and midbrain GnRH neurons. KT had no effect on any of the above neuronal parameters examined for salmon or seabream GnRH. Neither T3, EB, nor KT was effective to induce changes in midbrain chicken GnRH II mRNA concentrations, neuronal numbers, and neuronal size, indicating that an as yet unknown regulatory mechanism may operate midbrain GnRH neurons. T3 specifically suppressed the concentration of terminal nerve salmon GnRH mRNA, and EB significantly increased preoptic seabream GnRH neuronal numbers. These results are consistent with the hypothesis that thyroid hormone, by suppressing terminal nerve GnRH expression, promotes inhibition of sexual maturation. Furthermore, the failure of KT, a nonaromatizable androgen, to influence preoptic GnRH neurons emphasizes that an estrogenic pathway, at the onset of sexual maturation, is responsible for the recruitment of additional preoptic GnRH neurons that are fundamental to reproduction and behavior. PMID- 10803570 TI - Molecular and electrophysiological evidence for a GABAc receptor in thyrotropin secreting cells. AB - In the pituitary, GABA regulates the release of several hormones via different receptors. GABA(C) receptors are heterooligomers that differ from GABA(A) receptors in that they contain p-subunits and are insensitive to bicuculline. However, molecular and functional evidence for the presence of GABA(C) receptors outside the retina has yet to be established. The present work was performed on guinea pig and rat pituitaries. Both Northern blot and RT-PCR analysis showed that, although rho1- and rho2-subunits were expressed at similar levels in the rat retina, rho1 messenger RNA (mRNA) was enriched, relative to rho2 mRNA in the rat pituitary. Northern blot experiments also showed that, in the pituitary, rho1 and rho2 mRNAs are shorter in size than those expressed in the retina. The use of a subunit-specific antibody revealed colocalization of rho1-subunit and anti-TSH labeling on rat pituitary sections. TSH guinea pig pituitary cells were also labeled with a rho-subunit antiserum. Moreover, whole-cell patch clamp on single guinea pig TSH cells showed that GABA induced a bicuculline-insensitive Cl- current. In contrast to the Cl- current generated by GABA(C) receptors in the retina, the bicuculline-insensitive Cl- currents in TSH cells quickly desensitized. These results suggest that a novel GABA(C) receptor may regulate TSH secretion and that the structure and/or biochemical regulation of this pituitary receptor is different from that found in the retina. PMID- 10803571 TI - Thyroid hormone down-regulates neural cell adhesion molecule expression and affects attachment of gonocytes in Sertoli cell-gonocyte cocultures. AB - Contact-mediated interactions between Sertoli cells and gonocytes are important for testicular development. Specifically, down-regulation of neural cell adhesion molecule (NCAM)-based intercellular adhesion during postnatal maturation is likely to be important for appropriate differentiation of testicular cells. Besides NCAM, P-cadherin is also present in neonatal testicular cords, at least in mice, and seems to disappear from the seminiferous epithelium after the first postnatal week. Another factor known to be important in regulating development of the neonatal testis is thyroid hormone (T3). T3 is involved in control of Sertoli cell proliferation and differentiation. Therefore, we examined the effect(s) of T3 on adhesive factors found within the testis using Sertoli cells and gonocytes isolated from neonates and maintained in coculture. T3 (100 nM) down-regulated NCAM expression in vitro, as assessed by Western blotting and immunofluorescent staining. This contrasted with the continued expression of NCAM in cultures without added T3 but mimicked the disappearance of NCAM from the neonatal rat testis in vivo. In addition, Western analysis confirmed that P-cadherin is highly expressed in the developing rat testes, as it is in those of mice. We found that P-cadherin is strongly expressed in gonocytes and weakly expressed in Sertoli cells. Moreover, unlike NCAM, P-cadherin expression diminishes with time in vitro in the absence of added hormones. In parallel with our observations for NCAM, expression of P-cadherin was also apparently decreased by T3 (100 nM). Subsequent quantitative analyses of cultures exposed to a range of T3 levels (0.1-100 nM) indicated that T3 causes detachment of many gonocytes in a dose- and time dependent manner (approximately 80% detached at 100 nM). In addition, Western blotting indicated that lower concentrations of T3 down-regulate NCAM but not P cadherin. From this we conclude that the apparent decrease in P-cadherin induced by 100 nM T3 and detected on Western blots reflects loss of gonocytes. In contrast, even low levels of T3 appear to down-regulate NCAM production before any significant detachment of gonocytes. Finally, low levels of T3 that did not affect numbers of adherent Sertoli cells nevertheless caused detachment of gonocytes. Thus, our observations identify T3 as a regulator of NCAM expression in neonatal testicular cells and as a modifier of gonocyte/Sertoli cell adhesion in vitro. PMID- 10803572 TI - An aged rat model of partial androgen deficiency: prevention of both loss of bone and lean body mass by low-dose androgen replacement. AB - The aim of this study was to evaluate the effects of different doses of androgen replacement, both on body composition and bone, in an aged male orchidectomized rat model. Testosterone was administered by 0.5, 1, and 2.5-cm sc SILASTIC implants (release of, respectively, 11.5, 23, and 55 microg/day) to aged (12 months old, +/- 550 g) male orchidectomized Wistar rats during a 15-week experimental period. T 0.5 only partially prevented decrease of ventral prostate and seminal vesicle weight, compared with an intact group that received an empty implant (Intact). The 1-cm implant (T 1) completely prevented decrease of both seminal vesicles and ventral prostate weight. The 2.5-cm implant (T 2.5) was clearly supraphysiological, as demonstrated by significant hypertrophy of both androgen-sensitive organs. Serum testosterone was lower in T 0.5 and T 1 (0.38 +/ 0.06 ng/ml and 0.92 +/- 0.06 ng/ml, respectively) and higher in T 2.5 (2.4 +/- 0.28. ng/ml), compared with both Intact (1.6 +/- 0.23 ng/ml) and the baseline group(1.6 +/- 0.11 ng/ml). As expected, orchidectomized rats that received an empty SILASTIC implant had significantly lower bone mineral content (-7.9%), apparent density (-5.7%), and lean body mass (-10.8%), as measured by dual-energy x-ray absorptiometry, without significant changes in body weight and fat mass, compared with Intact. Also, cancellous (-50.3%) and cortical (-1.8%) volumetric density, as measured by peripheral quantitative computed tomography, were decreased in the tibia. Bone turnover, as measured by serum osteocalcin and urinary deoxypyridinoline excretion, was increased in orchidectomized rats that received an empty SILASTIC implant. T 0.5 prevented all changes, not only in bone mineral content, density, and turnover but also in lean body mass. Moreover, there were no significant differences, for all these parameters, between the different doses of testosterone replacement. In conclusion, low-dose androgen replacement does not lead to lower bone mineral density, higher bone turnover, and lower lean body mass in aged male rats, whereas complete androgen deficiency does. Therefore, the threshold concentration of testosterone necessary for prevention of both bone and lean body mass loss in aged male rats is clearly lower than for prostate and seminal vesicles. PMID- 10803573 TI - Growth-associated protein-43 messenger ribonucleic acid expression in gonadotropin-releasing hormone neurons during the rat estrous cycle. AB - We have shown previously at the ultrastructural level that morphological changes occur in the external zone of the median eminence allowing certain GnRH nerve terminals to contact the pericapillary space on the day of proestrus. The present study was designed to determine whether the intrinsic determinant of neuronal outgrowth, growth-associated protein-43 (GAP-43), was expressed in GnRH neurons of adult female rats, and whether its expression varied throughout the estrous cycle. To accomplish this, we perfusion-fixed groups of adult female rats at 0800 and 1600 h on diestrous day 2 (diestrous II), at 0800 h and 1600 h on proestrus, and at 0800 and 1600 h on estrus (n = 4 rats/group) and used double labeling in situ hybridization and quantification to compare the levels of GAP-43 messenger RNA (mRNA) in cells coexpressing GnRH mRNA. GnRH mRNA was detected with an antisense complementary RNA (cRNA) probe labeled with the hapten digoxigenin, whereas the GAP-43 cRNA probe was labeled with 35S and detected by autoradiography. In addition, GAP-43 protein was identified with immunohistochemistry in the median eminence. The results show that many GnRH neurons expressed GAP-43 mRNA and that GAP-43 protein was present in many GnRH axon terminals in the outer layer of the median eminence. The number of GnRH neurons expressing GAP-43 mRNA was significantly higher on proestrus (64 +/- 5%) than on diestrous II (40 +/- 2%; P < 0.001) or on estrus (45 +/- 8%; P < 0.05), and the GAP-43 mRNA levels in GnRH neurons also varied as a function of time of death during the estrous cycle. The GAP-43 mRNA levels in GnRH neurons were higher on proestrus and estrus than on diestrous II (P < 0.05). These data show that 1) GAP-43 is expressed in adult GnRH neurons; 2) GAP-43 mRNA expression in GnRH neurons fluctuates during the estrous cycle; and 3) GAP-43 mRNA content in GnRH neurons is highest on the day of proestrus, before and during the onset of the LH surge. These observations suggest that the increased GAP-43 mRNA expression in GnRH neurons on the day of proestrus could promote the outgrowth of GnRH axon terminals to establish direct neurovascular contacts in the external zone of the median eminence and thus facilitate GnRH release into the pituitary portal blood. PMID- 10803574 TI - Role of nitric oxide and cyclic guanosine 3',5'-monophosphate in the estrogen regulation of cervical epithelial permeability. AB - Treatment of cultured human cervical epithelia on filters with 17beta-estradiol increases paracellular permeability in a time- and dose-related manner (EC50, 1.1 nM). The objective of the present study was to understand the molecular mechanisms of estrogen action. In cultured human cervical epithelial cells the nitric oxide (NO) donors sodium nitroprusside (SNP) and N-[ethoxycarbonyl]-3-[4 morpholinyl]sydnoneimine (SIN-I) and the cell-permeable cGMP analog 8-bromo-cGMP (8-Br-cGMP) increased paracellular permeability. In estrogen-treated cells SNP and 8-Br-cGMP increased permeability to a lesser degree than in estrogen-deprived cells, suggesting that NO and cGMP mediate the effect of estrogen on permeability. Tamoxifen blocked the estrogen-induced increase in permeability, but it had no effect on increases in permeability that were induced by SNP or by 8-Br-cGMP. LY-83583 (blocker of guanylate cyclase) attenuated the effect of SNP, whereas KT-5823 (blocker of cGMP-dependent protein kinase) abrogated the effects of both SNP and 8-Br-cGMP. Treatment with 17beta-estradiol increased NO release and cellular cGMP in a dose-related manner (EC50, approximately 1 nM), and the effects were inhibited by tamoxifen. Treatment with SNP increased cGMP maximally, even in estrogen-deficient cells. LY-83583 blocked the estrogen-induced increase in cGMP, but neither LY-83583 nor KT-5823 had a significant effect on the estrogen-induced increases in NO release and cellular cGMP. The NO synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester decreased NO release, and pretreatment of cells with L-arginine reversed the effect. Cultured human cervical epithelial cells express messenger RNA for the NOS isoforms endothelial NOS (ecNOS), brain NOS, and inducible NOS. 17beta-Estradiol up-regulated ecNOS messenger RNA, and tamoxifen blocked the effect. Based on these results we suggest that the effect of estradiol on permeability involves four signaling steps: 1) activation of estrogen receptors, 2) increase in ecNOS transcription and up-regulation of NO activity, 3) NO activation of guanylate cyclase and increase in cGMP, and 4) cGMP activation of cGMP-dependent protein kinase. PMID- 10803575 TI - Expression of vascular endothelial growth factors and their receptors during osteoblast differentiation. AB - Endochondral bone formation is regulated by systemically and locally acting growth factors. A role for vascular endothelial growth factor (VEGF) in this process has recently been proposed, because inactivation of VEGF inhibits endochondral bone formation via inhibition of angiogenesis. Despite the known effect of VEGF as specific endothelial growth factor, its effects on osteoblast differentiation have not been studied. We, therefore, examined the expression of VEGF-A, -B, -C, and -D and their receptors in a model of osteoblast differentiation using the mouse preosteoblast-like cell line KS483. Early in differentiation, KS483 cells express low levels VEGF-A, -B, and -D messenger RNA, whereas during mineralization, KS483 cells express high levels. In addition, expression of the VEGF receptors, VEGFR1, VEGFR2, and VEGF165R/neuropilin, coincided with expression of their ligands, being maximally expressed during mineralization. VEGF-A production during osteoblast differentiation was stimulated by insulin-like growth factor I that enhances osteoblast differentiation and was inhibited by PTH-related peptide that inhibits osteoblast differentiation. Furthermore, continuous treatment of KS483 cells with recombinant human VEGF-A stimulated nodule formation. Although treatment of KS483 cells with soluble FLT1, an agent that blocks binding of VEGF-A and -B to VEGFR1, did not inhibit nodule formation, this observation does not exclude involvement of VEGFR2 in the regulation of osteoblast differentiation. As it is known that VEGF-A, -C, and -D can act through activation of VEGFR2, other isoforms might compensate for VEGF-A loss. The expression pattern of VEGFs and their receptors shown here suggests that VEGFs play an important role in the regulation of bone remodeling by attracting endothelial cells and osteoclasts and by stimulating osteoblast differentiation. PMID- 10803576 TI - Cannabinoid CB1 receptor-mediated inhibition of prolactin release and signaling mechanisms in GH4C1 cells. AB - The GH4C1 cell line was used to study the cellular mechanisms of cannabinoid mediated inhibition of PRL release. Cannabinoid CB1 receptor activation inhibited vasoactive intestinal polypeptide- and TRH-stimulated PRL release, but not its basal secretion. The cannabinoid-mediated inhibition of TRH-stimulated PRL release was reversed by the CB1 receptor-specific antagonist, SR141,716A, and was abolished by pertussis toxin pretreatment, indicating that G alpha subunits belonging to the G(i)alpha and G(o)alpha family were involved in the signaling. Photoaffinity labeling using [alpha-32P] azidoaniline GTP showed that cannabinoid receptor stimulation in cell membranes produced activation of four G alpha subunits (G(i)alpha2, G(i)alpha3, G(o)alpha1, and G(o)alpha2), which was also reversed by SR141,716A. The CB1 receptor agonists, WIN55,212-2 and CP55,940, inhibited cAMP formation and calcium currents in GH4C1 cells. The subtypes of calcium currents inhibited by WIN55,212-2 were characterized using holding potential sensitivity and calcium channel blockers. WIN55,212-2 inhibited the omega-conotoxin GVIA (Conus geographus)- and omega-agatoxin IVA (Aigelenopsis aperta)-sensitive calcium currents, but not the nisoldipine-sensitive calcium currents, suggesting the inhibition of N- and P-type, but not L-type, calcium currents. Taken together, the present findings indicate that CB1 receptors can couple through pertussis toxin-sensitive G alpha subunits to inhibit adenylyl cyclase and calcium currents and suppress PRL release from GH4C1 cells. PMID- 10803577 TI - CCAAT/enhancer-binding protein-alpha is a component of the growth hormone regulated network of liver transcription factors. AB - GH regulates gene expression by modulating the concentration or activity of transcription factors. To identify transcription factors that mediate the effects of GH in liver we analyzed the promoter of the gene coding for hepatocyte nuclear factor-6 (HNF-6), whose expression in liver is stimulated by GH. In protein-DNA interaction studies and in transfection experiments, we found that the liver enriched transcription factor CCAAT/enhancer-binding protein-alpha (C/EBPalpha) binds to the hnf6 gene and inhibits its expression. This inhibitory effect involved an N-terminal subdomain of C/EBPalpha and two sites in the hnf6 gene promoter. Using liver nuclear extracts from GH-treated hypophysectomized rats, we found that GH induces a rapid, transient decrease in the amount of C/EBPalpha protein. This GH-induced change is concomitant with the transient stimulatory effect of GH on the hnf6 gene. Stimulation of the hnf6 gene by GH therefore involves lifting of the repression exerted by C/EBPalpha in addition to the known GH-induced stimulatory effects of STAT5 (signal transducer and activator of transcription-5) and HNF-4 on that gene. Our data provide further evidence that GH controls a network of liver transcription factors and show that C/EBPalpha participates in this process. PMID- 10803578 TI - Coactivator and corepressor gene expression in rat cerebellum during postnatal development and the effect of altered thyroid status. AB - Thyroid hormone (TH) plays an important role in the postnatal development of the rodent cerebellum, particularly within the first 2 weeks of postnatal life. This action is exerted through the regulation of specific genes during development and is mediated by coactivator and corepressor proteins that determine transcriptional repression or activation, respectively. Thus, we hypothesized that the effect of TH on rodent cerebellar development could be influenced by the relative amounts of coactivator and corepressor proteins in vivo. These ratios might be modulated in an age-specific manner and/or by hormones to generate the "critical period" of TH action. To examine this hypothesis, we cloned rat complementary DNA fragments corresponding to coactivators (SRC1, TIF2 and TRAM1) and corepressors (N-CoR and SMRT), and studied the ontogenic changes in their corresponding messenger RNAs in rat cerebellum of normal and hypothyroid rats during postnatal development, using a RNase protection assay. We found an increased expression of SRC1 and TIF2, as well as of N-CoR, during rat cerebellar development but no change in the expression of SMRT and TRAM1 genes. However, thyroid hormone status did not affect the expression of coactivator and corepressor genes in the cerebellum. These results indicate that coactivator and corepressor messenger RNAs exhibit differential expression through cerebellar development but are not regulated by TH during this period. PMID- 10803579 TI - Androgen-induced growth inhibition of androgen receptor expressing androgen independent prostate cancer cells is mediated by increased levels of neutral endopeptidase. AB - Androgen-mediated growth repression of androgen-independent prostate cancer (AIPC) cells has been reported in androgen-independent PC-3 cells overexpressing the androgen receptor, and in androgen-independent derivatives of LNCaP cells that develop following prolonged culture in androgen-free media. Using two models of AIPC, PC3/AR cells and LNCaP-OM1 cells, a subclone of LNCaP cells derived by prolonged culturing in charcoal-stripped media, we investigated whether expression of neutral endopeptidase 24.11 (NEP), a cell-surface peptidase that cleaves and inactivates neuropeptides implicated in the growth of AIPC, is induced by androgen, and whether NEP contributes to the observed androgen mediated growth repression. These cell lines each express high levels of androgen receptor. Culturing in dihyrotestosterone (DHT) resulted in a 30-56% (PC3) and 35 43% (LNCaP-OM1) decrease in cell number over 7 days concomitant with a significant increase in NEP enzyme specific activity. Northern analysis detected an increase in NEP transcripts following DHT treatment in PC3/AR cells. The addition of the NEP enzyme inhibitor phosphoramidon to PC3 and LNCaP-OM1 or the NEP competitive inhibitor CGS 24592 to LNCaP-OM1 blocked the increase in NEP enzyme activity and reversed the DHT-induced growth inhibition. Neither phosphoramidon or CGS 24592 alone inhibited cell growth. Furthermore, the reversal of growth inhibition in LNCaPOM1 cells was dose dependent on the concentration of CGS 24592. These data indicate that androgen-induced growth repression of AIPC cells PC3 and LNCaP-OM1 results in part from androgen-induced expression of NEP in these cells. PMID- 10803580 TI - Effect of osmolarity on aldosterone production by rat adrenal glomerulosa cells. AB - The effect of osmotic changes on aldosterone production, [Ca2+]i and voltage gated Ca2+ currents, was studied in cultured rat glomerulosa cells. Alteration of osmolarity by sucrose addition in the 250-330 mosM range did not influence aldosterone production per se, but it substantially affected K+-stimulated aldosterone production. Hyposmosis markedly increased the hormone response evoked by raising [K+] from 3.6 to 5 mM, whereas hyperosmosis had a mild decreasing effect. Cytoplasmic [Ca2+]i, measured in single glomerulosa cells, did not show detectable change in response to either hyposmotic or hyperosmotic exposure, but the [Ca2+]i signal evoked by elevation of [K+] to 5 mM was augmented in hyposmotic solution. The osmosensitivity of the transient (T)-type and long lasting (L)-type voltage-gated Ca2+ currents was studied using the nystatin perforated voltage-clamp technique. Lowering osmolarity to 250 mosM significantly increased the amplitude of the T-type current, and it had a transient augmenting effect on L-type current amplitude. Hyperosmotic solution (330 mosM) reduced L type current amplitude but did not evoke significant change in T-type current. These results indicate that the responsiveness of rat glomerulosa cells to physiological elevation of [K+] is remarkably influenced by changes in osmolarity by means of modulating the function of voltage-gated Ca2+ channels. PMID- 10803581 TI - Expression of estrogen receptor alpha and beta in the rhesus monkey corpus luteum during the menstrual cycle: regulation by luteinizing hormone and progesterone. AB - There are conflicting reports on the presence or absence of estrogen receptor (ER) in the primate corpus luteum, and the discovery of a second type of estrogen receptor, ERbeta, adds an additional level of complexity. To reevaluate ER expression in the primate luteal tissue, we used semiquantitative RT-PCR based assays and Western blotting to assess ERalpha and beta messenger RNA (mRNA) and protein levels in corpora lutea (n = 3/stage) obtained from adult female rhesus monkeys at early (days 3-5), mid (days 6-8), mid-late (days 10-12), and late (days 14-16) luteal phase of the natural menstrual cycle. ERalpha mRNA levels did not vary across the stages of the luteal phase, and ERalpha protein was not consistently detected in luteal tissues. However, ERbeta mRNA and protein levels were detectable in early and mid luteal phases and increased (P < 0.05) to peak levels at mid-late luteal phase before declining by late luteal phase. To determine if ERbeta mRNA expression in the corpus luteum is regulated by LH, monkeys received the GnRH antagonist antide either alone or with 3 daily injections of LH to simulate pulsatile LH release. Treatment with antide alone or concomitant LH administration did not alter luteal ERbeta mRNA levels. When monkeys also received the 3beta-hydroxysteroid dehydrogenase inhibitor trilostane to reduce luteal progesterone production, luteal ERbeta mRNA levels were 3-fold higher (P < 0.05) than in monkeys receiving antide + LH only. Replacement of progestin activity with R5020 reduced luteal ERbeta mRNA levels to those seen in animals receiving antide + LH. Thus, there is dynamic ERbeta expression in the primate corpus luteum during the menstrual cycle, consistent with a role for estrogen in the regulation of primate luteal function and life span via a receptor (ERbeta)-mediated pathway. Increased ERbeta expression in the progestin depleted corpus luteum during LH exposure suggests that the relative progestin deprivation experienced by the corpus luteum between LH pulses may enhance luteal sensitivity to estrogens during the late luteal phase of the menstrual cycle. PMID- 10803582 TI - Expression of myometrial activation and stimulation genes in a mouse model of preterm labor: myometrial activation, stimulation, and preterm labor. AB - Myometrial contractions of labor result from an increase in myometrial activation and stimulation. Activation develops through the expression of contraction associated proteins (CAPs), including oxytocin receptors (OTR), connexin-43 (Cx 43), and prostaglandin F2 alpha, receptors (FP). Stimulation involves increases in contractile agonists including prostaglandin E2 (PGE2) and prostaglandin F2 alpha. (PGF2 alpha) that may result from increases in prostaglandin endoperoxide H synthase (PGHS)-2. A mouse model of preterm birth was used to study gene expression involved in myometrial activation and stimulation. To induce preterm birth, pregnant C57BL/6J mice were intubated with 6 g/kg ethanol on gestational day 16 and were killed every 6 h from treatment until birth. RIA was used to measure uterine PGE2 and PGF2 alpha, while PGHS-2, OTR, Cx-43, and FP messenger RNA levels were measured by ribonuclease protection assay. Increases in CAP mRNA were associated with term and preterm birth. There were differences in stimulation effectors associated with preterm and term birth. Uterine PGF2 alpha values were increased only at the time of term birth, but PGE2 was elevated during both preterm and term labor. These data suggest that existing levels of PGF2 alpha are sufficient for preterm birth when CAP expression is increased, but term labor requires increases in PGE2, PGF2alpha, and CAPs. The PGHS-2 messenger RNA expression pattern suggests that it is a CAP. PMID- 10803583 TI - 3,5-diiodo-L-thyronine regulates glucose-6-phosphate dehydrogenase activity in the rat. AB - Thyroid hormones influence the activity of lipogenic enzymes such as malic enzyme (ME) and glucose-6-phosphate dehydrogenase (G6PD). The effect of T3 on ME is exerted at the transcriptional level, but it is unclear if its effect on G6PD is also nuclear mediated. Furthermore, other iodothyronines that have been shown to possess biological activity (such as diiodothyronines) could contribute to this enzyme's regulation. In this study the effects of 3,5-diiodothyronine (T2) on the aforementioned enzymes were examined and compared with those of T3. Rats made hypothyroid by propylthiouracil and iopanoic acid treatment were used throughout. Enzyme activities were determined spectrophotometrically, and G6PD messenger RNA (mRNA) expression was analyzed by Northern blotting using a human G6PD complementary DNA probe. Injections of T2 to hypothyroid animals significantly enhanced the activity of both enzymes. The effect of T2 on ME was nuclear mediated and mimicked the effect of T3. The effects of T2 and T3 on G6PD differed. Injection of T3 into hypothyroid rats induced an increase in both enzyme activity and G6PD mRNA expression, indicating a nuclear-mediated effect. The effect of T2 on G6PD activity, on the other hand, was not nuclear mediated. The injection of T2 into hypothyroid animals did not change G6PD mRNA expression, and the strong increase in the enzyme's activity (from +70% to +300%) was unaffected by simultaneous injection of protein synthesis inhibitors. As the lowest dose of 1 microg T2/100 g BW affects G6PD activity 3-5 times more than the same dose of T3, these data provide the first evidence that T2 is a factor capable of regulating G6PD activity. PMID- 10803584 TI - Gonadotropin-releasing hormone analogs stimulate and testosterone inhibits the recovery of spermatogenesis in irradiated rats. AB - We investigated the effects of GnRH analogs, different doses of testosterone (T), an androgen receptor antagonist (flutamide), and combinations of these on the recovery of spermatogenesis after irradiation. Treatment with a GnRH agonist (Lupron) for 10 weeks after irradiation reduced the intratesticular T concentration (ITT) to 4% of that in irradiated rats and serum FSH to undetectable levels without altering serum LH levels. Injection of a GnRH antagonist (Cetrorelix) at 3 weeks after irradiation suppressed LH, FSH, and ITT to <7%, 32%, and 10%, respectively, of levels in irradiated-only rats within 2 weeks; suppression was maintained for approximately 3 to 4 weeks. The percentage of tubules with differentiated germ cells (repopulation index, RI) was <0.6% at weeks 10 to 20 after irradiation. Spermatogenic recovery was induced by both the GnRH agonist (RI = 58% at week 10; 91% at week 20) and antagonist (RI = 70% at week 13). There was a dose-dependent suppression of testicular germ cell repopulation when T was combined with GnRH analogs. The ability of T to abolish the spermatogenic stimulatory effect of the GnRH antagonist was evident by the similar RI obtained for irradiated rats given antagonist + T or T alone. This suppression of GnRH-induced recovery of spermatogenesis by T could be reversed by flutamide. The RI best correlated with the degree of ITT suppression. In ITT suppressed rats, the RI also showed an inverse correlation with serum T levels. Thus, T and/or its androgenic metabolites either directly or indirectly inhibit spermatogenic recovery after irradiation through an androgen receptor-mediated process. In addition, there was a close negative correlation between RI and FSH levels, and hence, a spermatogenic inhibitory role for FSH in the irradiated rats cannot be ruled out. PMID- 10803585 TI - The gp130 cytokines interleukin-11 and ciliary neurotropic factor regulate through specific receptors the function and growth of lactosomatotropic and folliculostellate pituitary cell lines. AB - Two of the most potent cytokines regulating anterior pituitary cell function are leukemia inhibitory factor and interleukin-6 (IL-6), which belong to the cytokine receptor family using the common gp130 signal transducer. We studied the actions of two other members of this family, IL-11 and ciliary neurotropic factor (CNTF), on folliculostellate (FS) cells (TtT/GF cell line) and lactosomatotropic cells (GH3 cell line). The messenger RNA (mRNA) for the alpha-chain specific for the IL 11 receptor (1.7 kb) and CNTF receptor (2 kb) are expressed on both cell types. In addition, we detected CNTF receptor mRNA in normal rat anterior pituitary cells. IL-11 (1.25-5 nM) dose dependently stimulated the proliferation of FS cells. CNTF, at doses from 0.4-2 nM, also significantly stimulated the growth of these cells. In addition, both cytokines significantly stimulated proliferation of lactosomatotropic GH3 cells, and CNTF stimulated hormone production (GH and PRL) at 24 h by these cells. At 16-72 h, IL-11 stimulates the secretion of the angiogenic factor vascular endothelial growth factor by FS cells. In addition, both GH3 and FS cells express CNTF mRNA. These data suggest that IL-11 and CNTF may act as growth and regulatory factors in anterior pituitary cells. PMID- 10803586 TI - Regulation of gonadotropin-releasing hormone and its receptor gene expression by 17beta-estradiol in cultured human granulosa-luteal cells. AB - There is evidence that GnRH and its binding sites are expressed in numerous extrapituitary tissues, including the primate ovary. However, the factors that regulate ovarian GnRH and its receptor (GnRH-R) remain poorly characterized. Since gonadal steroids are key regulators of ovarian functions, the present study investigated the role of 17beta-estradiol (E2) in regulating GnRH and GnRH-R messenger RNA (mRNA) from human granulosa-luteal cells (hGLCs). RT-PCR was used to isolate the ovarian GnRH-R transcript equivalent to the full-length coding region in the pituitary from hGLCs. Sequence analysis revealed that the ovarian GnRH-R mRNA is identical to its pituitary counterpart. Basal expression studies indicated that GnRH and GnRH-R mRNA levels significantly increased with time in vitro, reaching levels of 160% and 170% on day 8 and 10 of culture, respectively (P < 0.05). Treatment with various concentrations of estradiol (1-100 nM) for 24 h resulted in a dose-dependent decrease (P < 0.05) in GnRH and GnRH-R mRNA levels. Time course studies indicated that short-term treatment (6 h) with E2 (1 nM) had no significant effect on GnRH mRNA levels, while long-term treatment (48 h) with E2 resulted in a 40% decrease (P < 0.001) in GnRH mRNA levels. In contrast, GnRH-R mRNA levels exhibited a biphasic pattern, such that a short-term treatment (6 h) with E2 increased GnRH-R mRNA levels by 20% (P < 0.05), whereas long-term treatment (48 h) resulted in a 60% decrease (P < 0.001) in GnRH-R expression in hGLCs. Cotreatment of estradiol and tamoxifen blocked the E2 induced-regulation of GnRH and its receptor mRNAs, indicating that the E2 effect was mediated through its receptor. In summary, our studies demonstrate that the ovary possesses an intrinsic GnRH axis that is regulated during luteinization in vitro, and that E2 is capable of regulating GnRH and its receptor in the human ovary. PMID- 10803587 TI - Complementary deoxyribonucleic acid cloning, gene expression, and ligand selectivity of a novel gonadotropin-releasing hormone receptor expressed in the pituitary and midbrain of Xenopus laevis. AB - We have cloned the full-length complementary DNA (cDNA) for a GnRH receptor from Xenopus laevis pituitary cDNA and determined its gene structure. The cDNA encodes a 368-amino acid protein that has a 46% amino acid identity to the human GnRH receptor. The X laevis GnRH receptor has all of the amino acids identified in the mammalian GnRH receptors as sites of interaction with the GnRH ligand. However, this receptor cDNA shares the same distinguishing structural features of the GnRH receptor that have been characterized from other nonmammalian vertebrates. These include the pair of aspartate residues in the transmembrane domains II and VII compared with the aspartate/asparagine arrangement in mammalian receptors, the amino acid PEY motif in extracellular loop III (SEP in mammals), and the presence of a carboxyl-terminal tail. Previous studies have reported that mammalian GnRH was equipotent to other naturally occurring GnRH subtypes in stimulating LH release from the amphibian pituitary. However, in this study we show that the X. laevis GnRH receptor has ligand selectivity for the naturally occurring GnRHs similar to other nonmammalian GnRH receptors. The order of potency of the GnRHs in stimulating inositol phosphate production in COS-1 cells transiently transfected with the X. laevis GnRH receptor cDNA was chicken GnRH II>salmon GnRH>mammalian GnRH. Transcripts of this GnRH receptor are expressed in the pituitary and midbrain of X. laevis. PMID- 10803588 TI - The role of neuropeptide Y in the progesterone-induced luteinizing hormone releasing hormone surge in vivo in ovariectomized female rhesus monkeys. AB - Progesterone induces a LHRH surge in estrogen-primed ovariectomized rhesus monkeys, with a concomitant increase in the pulse frequency of neuropeptide Y (NPY) release. However, the role for NPY in the positive feedback action of progesterone on LHRH release in primates is unknown. The present study examines the effect of an antisense oligodeoxynucleotide for NPY messenger RNA (AS NPY) on the progesterone-induced LHRH surge in vivo using push-pull perfusion. The AS NPY was directly infused into the stalk-median eminence (S-ME), whereas perfusates were collected for assessment of LHRH release. For a control, a scrambled oligodeoxynucleotide was infused. The results indicate that 1) the scrambled oligodeoxynucleotide did not interfere with the progesterone-induced LHRH surge, 2) whereas AS NPY blocked the progesterone-induced increase in LHRH release, and 3) no LHRH surges were induced by oil as a control for progesterone, but the AS NPY also reduced LHRH release in oil controls. These data suggest that 1) AS NPY infusion into the S-ME results in reduction in LHRH release; and 2) NPY release in the S-ME is important for the positive feedback effects of progesterone on LHRH release in estrogen-primed ovariectomized monkeys. PMID- 10803589 TI - Expression of GalR1 and GalR2 galanin receptor messenger ribonucleic acid in proopiomelanocortin neurons of the rat arcuate nucleus: effect of testosterone. AB - Previous studies have shown that galanin-containing fibers make synaptic contacts with POMC neurons in the arcuate nucleus. However, the ability of POMC neurons to express galanin receptors has never been assessed. The present study was designed to investigate whether POMC neurons express galanin receptor messenger RNA (mRNA) and whether testosterone could modulate galanin receptor gene expression. A dual labeling in situ hybridization histochemistry, using 35S-labeled (galanin receptors GalR1 or GalR2) and digoxigenin-labeled (POMC) riboprobes, was performed on brain sections from intact, castrated, and testosterone-replaced adult male rats. For analysis, the arcuate nucleus was divided into four rostro caudal areas. The results revealed that both GalR1 and GalR2 mRNAs were expressed in POMC neurons. Most POMC neurons expressing galanin receptor mRNAs were found in the rostral parts of the nucleus. Castration reduced the labeling density of galanin receptor mRNAs in POMC neurons, and testosterone prevented the effects of castration in all rostro-caudal subdivisions of the arcuate nucleus. Taken together, these data indicate that galanin can directly modulate the activity of POMC neurons, via an action on GalR1 or GalR2 receptors, particularly in the rostral-arcuate nucleus. In addition, testosterone can modulate the expression of GalR1 and GalR2. Because POMC neurons located in the rostral part of the nucleus are known to project preferentially to the preoptic area, POMC neurons expressing the galanin receptor genes may play an important role in the regulation of the GnRH neuroendocrine axis. PMID- 10803590 TI - The effect of a null mutation in the follicle-stimulating hormone receptor gene on mouse reproduction. AB - To investigate further brain-pituitary-gonadal interrelationships we have generated mice in which the gene encoding the FSH receptor has been disrupted. Female FSH receptor knockout (FSHRKO) mice were infertile. The ovaries were significantly reduced in size, with follicular development arrested at the preantral stage, but there was evidence of stromal hypertrophy. The vagina was imperforate, and the uterus was atrophic. There was no response to administration of PMSG. Inhibins A and B were undetectable in both the serum and gonads. Compared with those in control animals, serum concentrations of FSH and LH were significantly elevated in mutant females. The pituitary content of FSH, but not LH, was also significantly elevated. Estrogen administration in FSHRKO female mice suppressed serum LH levels to those seen in control mice, whereas FSH levels were reduced by only 50%. Male FSHRKO mice were fertile, although testis weight was significantly reduced. However, testicular inhibin A and B concentrations did not differ from those in normal littermates. Serum levels of FSH and LH were elevated in the null mutant male mice, whereas no differences were found in the pituitary content of these hormones. In conclusion, ovarian follicular development cannot progress beyond the preantral stage without FSH. In the absence of mature follicles ovarian estrogen remains low, and consequently accessory sex tissue growth and negative feedback regulation of gonadotropin secretion are severely compromised. In the male, however, inability to respond to FSH does not impair fertility, although testicular weight is reduced, and feedback regulation of pituitary gonadotropins and intratesticular paracrine interactions may be disturbed. PMID- 10803591 TI - Isolation of human type 2 deiodinase gene promoter and characterization of a functional cyclic adenosine monophosphate response element. AB - We analyzed the structure and function of the 5' flanking region of the human type 2 deiodinase (hD2) gene. Two major transcription start sites were identified at -470/-474 from the ATG. The 5' flanking region of hD2 gene efficiently directed transcription in transient transfection studies, using luciferase as reporter gene, in HEK 293 cells. Basal transcriptional activity was significantly reduced by deleting the region containing a canonical cAMP-responsive element (CRE) located -766/-759 from ATG. Forskolin treatment significantly increased luciferase activity in cells transfected with CRE-containing constructs. This effect was abolished in constructs that did not contain CRE or contained the mutagenized CRE. Northern blot analysis in JEG-3 cells revealed that the hD2 messenger RNA was markedly increased after stimulation with cAMP agonist. The electrophoretic mobility shift assay with hD2-CRE probe and HEK 293 nuclear extract showed the occurrence of a DNA-protein complex, which was competed by specific unlabeled oligonucleotides and supershifted by the anti-CREB and anti CRE modulator-1 antibodies. A-CREB, a dominant negative inhibitor of CREB, completely inhibited forskolin induction of the hD2 promoter. CREB protein, once cotransfected with hD2 promoter construct and pKA in F9 teratocarcinoma cells, which are unresponsive to cAMP, was able to stimulate the hD2 gene transcription. These results indicate the existence of a functional promoter within the 5' flanking region of hD2 gene which is characterized by the presence of a CRE. The specific involvement of CREB in the cAMP-mediated hD2 gene promoter induction also has been demonstrated. PMID- 10803592 TI - Cell-specific induction of sensitivity to ganciclovir in medullary thyroid carcinoma cells by adenovirus-mediated gene transfer of herpes simplex virus thymidine kinase. AB - Herpes simplex virus thymidine kinase (HSVtk) gene transfer followed by ganciclovir administration is a common strategy for experimental cancer therapy. To evaluate the feasibility of using the human calcitonin promoter to target medullary thyroid carcinoma (MTC), we developed adenovirus vectors containing Escherichia coli beta-galactosidase gene under the control of the CALC-I promoter (AdCTlacZ), or the human cytomegalovirus promoter (AdCMVlacZ). Beta-galactosidase activity driven by the CALC-I promoter was higher than by the CMV promoter in rat MTC cells after infection with adenovirus vectors. AdCTlacZ induced an equal or lower expression level of beta-galactosidase in TT (human MTC), T98G, Cos1, HepG2, and HeLa cells compared with AdCMVlacZ. To inhibit the growth of MTC cells, we developed two adenovirus vectors, AdCMVtk carrying HSVtk driven by the cytomegalovirus promoter and AdDCTtk containing a human CALC-I minigene under the control of the CALC-I promoter. HSVtk is fused to a portion of calcitonin coded in exon 4 to direct cell-specific regulation of splicing. All cell lines infected with AdCMVtk were rendered sensitive to ganciclovir, whereas T98G and Cos1 cells infected with AdDCTtk were not affected. Cell killing was also observed in HeLa, HepG2, rat MTC and TT cells infected with AdDCTtk. PMID- 10803593 TI - Transcription factor activator protein-2 is required for continued luteinizing hormone-releasing hormone expression in the forebrain of developing mice. AB - LHRH is the neuropeptide responsible for reproductive function. Prenatally, LHRH expression begins when neurons are in the olfactory pit and continues as these cells migrate into the brain. Thus, LHRH neurons maintain neuropeptide expression through very distinct environments. The regulatory interactions that control onset and continued expression of the LHRH phenotype are unknown. To begin to address this question primary LHRH neurons were removed from nasal explants at different ages. A complementary DNA (cDNA) subtraction screen was performed comparing a 3.5-days in vitro LHRH neuron [approximately embryonic day 15 (E15) in vivo] to two 10.5-days in vitro LHRH neurons (approximately postnatal day 1 in vivo). The transcription factor activator protein-2 (AP-2alpha) was differentially expressed and was present in the developmentally younger LHRH neuron. In vivo analysis revealed that LHRH neurons expressed AP-2 as they migrated across the cribriform plate and into the forebrain beginning on E13.5, but that coexpression of LHRH and AP-2 was no longer detected in postnatal day 1 animals. This suggested a regulatory role for AP-2 in LHRH neurons. Analysis of animals lacking AP-2alpha revealed a dramatic decrease in forebrain LHRH neurons between E13.5 and E14.5, correlating with normal onset of AP-2 expression in LHRH neurons as they entered the central nervous system. Nasal cells robustly expressing LHRH were still present on E 14.5. The continued presence of forebrain LHRH cells is proposed based on a second marker, galanin, and lack of increased apoptotic/necrotic cells in this region. A decrease in LHRH messenger RNA in forebrain neurons indicates regulation of LHRH occurred at the transcriptional or posttranscriptional level in mutant animals. These results indicate a developmentally restricted involvement of the transcription factor AP-2 in LHRH expression once the LHRH neurons have migrated into the forebrain, but before establishment of an adult-like distribution. PMID- 10803594 TI - Estradiol induces differential neuronal phenotypes by activating estrogen receptor alpha or beta. AB - Estrogens are female sex steroids that have a plethora of effects on a wide range of tissues. These effects are mediated through two well characterized intracellular receptors: estrogen receptor alpha and beta (ERalpha and ERbeta, respectively). Because of their high structural homology, it has been argued whether these two receptors may elicit differential biochemical events in estrogen target cells. Here we examine the effect of 17beta-estradiol-dependent activation of ERalpha and ERbeta on neurite sprouting, a well known consequence of this sex hormone action in neural cells. In SK-N-BE neuroblastoma cells transfected with ERalpha or ERbeta, 17beta-estradiol induces two distinct morphological phenotypes. ERalpha activation results in increased length and number of neurites, whereas ERbeta activation modulates only neurite elongation. By the use of chimeric receptors we demonstrate that the presence of both transcription activation functions located in the NH2-terminus and COOH-terminus of the two ER proteins are necessary for maintaining the differential biological activity reported. ERalpha-dependent, but not ERbeta-dependent, morphological changes are observed only in the presence of the active form of the small G protein Rac1B. Our data provide the first clear evidence that, in a given target cell, ERalpha and ERbeta may play distinct biological roles and support the hypothesis that 17beta-estradiol activates selected intracellular signaling pathways depending on the receptor subtype bound. PMID- 10803596 TI - Delineation of an antiapoptotic action of glucocorticoids in hepatoma cells: the role of nuclear factor-kappaB. AB - Glucocorticoids are primarily recognized for their profound anti-inflammatory actions and their ability to induce lymphocyte apoptosis. We report here that, in contrast to their effect on cells of the immune system, glucocorticoids suppress serum deprivation induced apoptosis of rat hepatoma (HTC) cells. Suppression of apoptosis in these cells occurs at physiological concentrations of glucocorticoid and is abrogated by the glucocorticoid antagonist RU486. Although HTC cells also express receptors for progesterone, estrogen, and thyroid hormone, ligands for these receptors fail to rescue these cells from programmed cell death. Because the sensitivity of cells to apoptotic stimuli is often regulated by the ratio of antiapoptotic to proapoptotic Bcl-2 family members, we analyzed the influence of glucocorticoids and induction of apoptosis by serum starvation on the expression of these proteins. Bcl-2, Bcl-xL, Bad, Bak, and Bax levels were not altered by either treatment. Mitochondrial function has recently been implicated as a critical early regulator of apoptosis in many cells including hepatocytes. Dexamethasone treatment blocked a decrease in this potential (delta psi(m)) during serum deprivation induced apoptosis in HTC cells, indicating an action of this hormone upstream of mitochondria. We also show that the induction of apoptosis in HTC cells is associated with a decrease in nuclear factor (NF) kappaB. Treatment with dexamethasone effectively blocked the loss of nuclear NF kappaB, suggesting that this hormone acts to suppress apoptosis of HTC cells via regulation of this nuclear transcription factor. This hypothesis was confirmed by transfection experiments that show that expression of a superrepressor of NF kappaB inhibits the ability of dexamethasone to rescue HTC cells from apoptosis induced by serum deprivation. PMID- 10803595 TI - Caspase-3 activation is required for Leydig cell apoptosis induced by ethane dimethanesulfonate. AB - Previous studies have shown that ethane dimethanesulfonate (EDS) causes the apoptotic death of Leydig cells. The molecular mechanism by which EDS elicits its effect remains uncertain. The present study tested the hypothesis that caspase-3 is involved in the EDS-induced death of rat Leydig cells. Leydig cells were isolated from adult Sprague Dawley at 3, 6, 12, or 24 h after the rats received an EDS injection. Low mol wt DNA fragments that are characteristic of apoptosis were evident by 12 h post-EDS, and the ladder pattern was more pronounced at 24 h. During this same time period, the number of terminal deoxynucleotidyltransferase-mediated deoxy-UTP-biotin nick end labeling (TUNEL) positive cells increased. Western blot analysis revealed that procaspase-3 was present only at low levels in control Leydig cells, and increased through 6 h post-EDS. By 12 h, procaspase-3 was reduced, whereas the cleaved, active caspase 3 forms appeared at 12 h and increased through 24 h post-EDS. Caspase-3 activity was blocked by caspase-3 inhibitor. In vitro, EDS treatment induced Leydig cell apoptosis. In the presence of cell-permeable caspase-3 inhibitor, however, apoptosis was significantly suppressed, providing further evidence for the involvement of caspase-3 in EDS-induced Leydig cell apoptotic death. Immunohistochemical analysis revealed weak staining for caspase-3 in the cytoplasm of control Leydig cells. From 12-24 h post-EDS, the time interval during which the active forms of caspase-3 appeared, caspase-3 immunoreactivity increased and became localized to the nuclei. Apoptosis and caspase-3 were colocalized in Leydig cells by a histological method that combined TUNEL and caspase-3 immunohistochemistry. In these studies, TUNEL-positive cells all exhibited intense nuclear caspase-3 immunoreactivity, whereas TUNEL-negative cells exhibited weak caspase-3 immunoreactivity in the cytoplasm. Taken together, these results indicate that Leydig cell apoptosis induced by EDS is mediated by caspase-3 activation, and suggest that the translocation of the active caspase-3 forms to the nucleus may be involved. PMID- 10803597 TI - Mrp4, a new mitogen-regulated protein/proliferin gene; unique in this gene family for its expression in the adult mouse tail and ear. AB - Mitogen-regulated proteins (also known as proliferin; mrp/plf) are nonclassical members of the PRL/GH family. They are expressed at high levels during midgestation when they are thought to induce angiogenesis and uterine growth. There are between four and six mrp/plf genes, and three different complementary DNAs have been cloned. Here we identify a fourth mrp/plf gene (mrp4) that we have cloned and characterized. MRP4 is 91% identical in amino acid sequence with the other MRP/PLF proteins but is missing two glycosylation sites that are present in the other forms. Consistent with the loss of two of three glycosylation sites, the expressed form of MRP4 has a lower apparent molecular weight compared with other MRP/PLFs. In vivo, mrp4 is expressed in the placenta and the adult skin. Expression of mrp4 messenger RNA peaks in the placenta on day 12. In the skin, mrp4 expression is specific to the ears and tails of mice. Our results suggest that, as well as having growth and angiogenic effects during pregnancy, the MRP/PLFs may have functions in nonreproductive tissues. Unique among the members of the mrp/plf family for its expression in the hair follicles of the tail and ear, MRP4 is expected to have a singular role in the growth and development of these follicles. PMID- 10803598 TI - Implantation and decidualization defects in prolactin receptor (PRLR)-deficient mice are mediated by ovarian but not uterine PRLR. AB - PRL and its homologs accomplish their biological effects through the PRL receptor (PRLR). We evaluated the expression and function of PRLR in the embryo and uterus during the periimplantation period because PRLR deficiency results in implantation failure. In wild-type mice, PRLR expression was localized to undecidualized stromal cells in the antimesometrial border on days 6-8 of pregnancy. A small population of PRLR-expressing cells was observed adjacent to the ectoplacental cone in the mesometrial stroma. Low levels of PRLR expression were also detected in the developing embryo on days 6-8. To determine the significance of PRLR expression in this distribution, we examined implantation and decidualization in PRLR-/- mice. Progesterone (P4) administration rescued infertility in PRLR-/- mice from the periimplantation period to midgestation. Artificially induced decidualization was absent in pseudopregnant PRLR-/- mice but was identical to wild-type in P4-treated PRLR-/- mice. Furthermore, wild-type and P4-treated PRLR-/- mice had similar expression of the implantation-specific genes, LIF, amphiregulin, HB-EGF, COX-1, COX-2, PPARdelta, Hoxa-10, cyclin-D3, VEGF, and its receptors, Flk-1 and neuropilin-1. Together, these results show that luteal P4 production via ovarian PRLR signaling is required for implantation and early pregnancy. The function of uterine PRLR remains unclear. However, the eventual loss of pregnancy in P4-treated PRLR-/- mice suggests that uterine PRLR may be essential for the support of late gestation. PMID- 10803599 TI - Enhanced growth of MCF-7 breast cancer cells overexpressing parathyroid hormone related peptide. AB - PTH-related peptide (PTHrP) is a secreted protein produced by breast cancer cells both in vivo and in vitro. Because of its structural similarity to PTH at the amino terminus, the two proteins interact with a common cell surface receptor, the PTH/PTHrP receptor. When overproduced by tumor cells, PTHrP enters the circulation, giving rise to the common paraneoplastic syndrome of humoral hypercalcemia of malignancy. Although initially discovered in malignancies, PTHrP is now known to be produced by most cells and tissues in the body. It acts as an autocrine and paracrine mediator of cell proliferation and differentiation, effects which are mediated via the PTH/PTHrP receptor. Recent evidence also has shown that, directly after translation, PTHrP is able to enter the nucleus and/or nucleolus and influence cell cycle progression and apoptosis. In this study, we have either overproduced PTHrP or inhibited endogenous PTHrP production in the breast cancer cell line, MCF-7. Overexpression of PTHrP was associated with an increase in mitogenesis, whereas inhibiting endogenous PTHrP production resulted in decreased cell proliferation. The overexpressed peptide targeted to the perinuclear space. In contrast, PTHrP interaction with the cell surface PTH/PTHrP receptor resulted in decreased cell proliferation in the same cell line. This latter effect is dependent on interaction with the receptor, in that exogenously added PTHrP moieties known not to interact with the receptor had no effect on cell growth. Furthermore, neutralization of added peptide with an anti-PTHrP antiserum completely abolished the growth inhibitory effects. In contrast, this antibody has no effect on the increased proliferation rate of the MCF-7 transfectants that overexpress PTHrP, compared with control cells. The net effect of autocrine/paracrine and intracrine effects of PTHrP in MCF-7 cells overproducing the peptide is accelerated cell growth. These findings have critical implications regarding the role of PTHrP in breast cancer, and they suggest that controlling PTHrP production in breast cancer may be useful therapeutically. PMID- 10803600 TI - GDNF and RET-gene expression in anterior pituitary-cell types. AB - Glial cell line-derived neurotrophic factor (GDNF) is expressed in many neuronal and non-neuronal tissues during development as well as in adult animals. GDNF signaling is mediated through a two-component system consisting of the so called GDNF receptor-alfa (GFRalpha1) which binds to GDNF. Thereafter this complex binds to and activates the tyrosine kinase receptor RET. In this work, for the first time, we have characterized the expression of both GDNF and RET in the anterior pituitary. First of all, RT-PCR analysis, Western blot and immunohistochemistry of the whole anterior pituitary showed that GDNF, GFRalpha1 and RET are expressed in this gland. Following double-immunofluorescence of consecutive sections we found GDNF immunoreactivity in most cell types, and it was most abundant in corticotrophs (55%), LH (59%) and FSH-producing cells (81%). In contrast, while the majority of somatotrophs (87%) were stained for RET, no positive immunostaining could be detected in other cell types. Taken together, this data indicate that gonadotrophs and corticotrophs are the main source of GDNF synthesized in the anterior pituitary and that the somatotrophs appears to be their target cell. This study provides direct morphological evidences that GDNF may well be acting in a paracrine-like fashion in the regulation of somatotroph cell growth and/or cell function. PMID- 10803601 TI - Potent inhibition of estrogen sulfotransferase by hydroxylated PCB metabolites: a novel pathway explaining the estrogenic activity of PCBs. AB - Polychlorinated biphenyls (PCBs) are persistent environmental pollutants which exert a variety of toxic effects in animals, including disturbances of sexual development and reproductive function. The estrogenic effects of PCBs may be mediated in part by hydroxylated PCB metabolites (PCB-OHs), but the mechanisms by which they are brought about are not understood. PCBs as well as PCB-Hs show low affinities for both alpha and beta estrogen receptor isoforms. In the present study we demonstrate that various environmentally relevant PCB-OHs are extremely potent inhibitors of human estrogen sulfotransferase, strongly suggesting that they indirectly induce estrogenic activity by increasing estradiol bioavailability in target tissues. PMID- 10803603 TI - Activin A release into the circulation is an early event in systemic inflammation and precedes the release of follistatin. AB - Recent evidence suggests a role for activin A, and its binding protein, follistatin, in inflammatory pathways. However, whether activin is released systemically during inflammation is not known. In this study, a release of activin A into the circulation occurred in sheep within 1 hour of injection of lipopolysaccharide. This rapid peak in activin A preceded the release of the key inflammatory cytokines, tumor necrosis factor-alpha and interleukin-6. Follistatin release into the circulation occurred some 4 hours after the peak of activin A and continued out to 24 hours from lipopolysaccharide treatment. These data are the first to document a circulatory response of activin A to an inflammatory stimulus, and together with previous findings, suggest that activin A may have both pro- and anti-inflammatory actions in regulating cytokine-driven pathways. PMID- 10803602 TI - Sodium-induced cardiac aldosterone synthesis causes cardiac hypertrophy. AB - High sodium intake causes cardiac hypertrophy independently of increases in blood pressure. Aldosterone is synthesized in extraadrenal tissues such as blood vessels, brain, and heart. Effects of 8 weeks of high sodium intake on cardiac aldosterone synthesis, as well as cardiac structure, mass, and aldosterone production, levels of mRNA coding for aldosterone synthase (CYP11B2) and the angiotensin II AT1 receptor, were studied in normotensive Wistar-Kyoto (WKY) rats. Isolated rat hearts were perfused for 2 hr, and the perfusate was analyzed by high-performance liquid chromatography and mass spectrometry. Aldosterone synthase activity was estimated from the conversion of [14C]deoxycorticosterone to [14C]aldosterone. Levels of mRNA for CYP11B2 and AT1 receptor were determined by competitive polymerase chain reactions. A high sodium intake for 8 weeks produced left ventricular hypertrophy without elevation of blood pressure. Plasma aldosterone concentrations and plasma renin concentrations were decreased by high sodium intake. Aldosterone production, activity of aldosterone synthase, and expression of mRNA for CYP11B2 and AT1 receptor were increased in hearts of rats with high sodium intake. These results suggest that high sodium intake increases cardiac aldosterone synthesis, which may contribute to cardiac hypertrophy independently of the circulating renin-angiotensin-aldosterone system. PMID- 10803604 TI - Prolactin releasing peptide (PrRP) stimulates luteinizing hormone (LH) and follicle stimulating hormone (FSH) via a hypothalamic mechanism in male rats. AB - Prolactin releasing peptide (PrRP) was originally isolated as an endogenous hypothalamic ligand for the hGR3 orphan receptor. It has been shown to release prolactin from dispersed pituitaries harvested from lactating female rats and only at very high doses in cycling females. PrRP is reported to have no effect on prolactin production from dispersed pituitary cells harvested from males. The CNS distribution of this peptide suggested a role for PrRP in the control of the hypothalamo-pituitary axis. The aim of this study was to examine the actions of PrRP (1-31) on circulating pituitary hormones following intracerebroventricular (ICV) injection in male rats and to investigate the mechanism of PrRP's effect by measurement of hypothalamic releasing factors in vitro. In our experiments, PrRP (1-31) did not release LH, FSH, TSH, growth hormone or prolactin directly from dispersed male pituitary cells in vitro. We have shown for the first time that following ICV injection of PrRP (1-31) 5 nmol there was a highly significant simulation of plasma LH that began at 10 minutes and was maintained over the course of the experiment (at 60 minutes PrRP 5 nmol 2.2 +/- 0.2 vs. saline 0.5 +/ 0.1 ng/ml, p<0.001). Plasma FSH increased at 20 minutes following ICV injection (PrRP 5nmol 10.8 +/- 2.0 ng/ml vs. saline 5.1 +/- 0.5, p<0.01). Total plasma testosterone increased at 60 minutes post injection (PrRP 5nmol 9.2 +/- 1.6 vs. saline 3.5 +/- 0.6 nmol/l, p<0.01). There was no significant alteration in plasma prolactin levels. PrRP significantly increased the release of LHRH from hypothalamic explants in vitro (PrRP 100nmol/l 180.5 +/- 34.5% of the basal secretion, p<0.05). PrRP (100nmol/l) also increased the following hypothalamic peptides involved in the control of pituitary hormone release, vasoactive intestinal peptide (VIP) 188.1 +/- 24.6% and galanin 153.8 +/- 13.0% (both p<0.001 vs. basal secretion) but had no effect on orexin A secretion. These results suggest a role for PrRP in the control of gonadotrophin secretion acting via a hypothalamic mechanism involving the release of LHRH. PMID- 10803605 TI - High throughput fluorescence polarization: a homogeneous alternative to radioligand binding for cell surface receptors. AB - High throughput fluorescence polarization (FP) assays are described that offer a nonradioactive, homogeneous, and low-cost alternative to radioligand binding assays for cell surface receptors (G protein-coupled receptors and ligand-gated ion channels). FP assays were shown to work across a range of both peptide (vasopressin V1a and delta-opioid) and nonpeptide (beta1-adrenoceptor, 5 hydroxytryptamine3) receptors. Structure-activity relationships were investigated at beta1-receptors and were found to be consistent with radioligand binding assays. FP was shown to tolerate up to 5% DMSO with no loss in sensitivity or signal window. From a random set of 1,280 compounds, 1.9% were found to significantly interfere with FP measurement. If fluorescent or quenching compounds were eliminated (3% of all compounds), less than 0.4% of compounds were found to interfere with FP measurement. Assays could be run in 384-well plates with little loss of signal window or sensitivity compared to 96-well plate assays. New advances in FP measurement have therefore enabled FP to offer a high throughput alternative to radioligand binding for cell surface receptors. PMID- 10803606 TI - Miniaturization of a functional transcription assay in yeast (human progesterone receptor) in the 384- and 1536-well plate format. AB - Miniaturization of high throughput screening assays to high-density microplate formats (384 or 1536 wells) is currently the focus of research activity in modern drug discovery facilities. In this article, we describe the adaptation of a fluorescence-based functional transcription assay in yeast for assessing modulators of human progesterone receptor to the 384- and 1536-well microplate format, comparing the experimental results to those obtained in the well established 96-well format. The experiences gained from the optimization of the liquid-handling procedures and the miniaturization of an enzyme assay (beta galactosidase) were implemented. Thus optimized pipetting protocols were developed to perform a reporter gene assay in yeast in microplate formats of higher density. In the functional transcription assay in yeast, the reporter gene expression showed the expected dependence on the ligand's dose and affinity in principle in all three microplate formats. For the first time, this assay system has been established in the 1536-well microplate format using CyBi-Well 96/384/1536 as the liquid-handling unit. The comparison of the signal:background ratios showed a lower sensitivity of the assay in the microplate formats of higher density. This study is an example of a successful miniaturization of a yeast cell-based assay to high-density plate formats on the basis of a careful adaptation procedure and optimized liquid-handling conditions. PMID- 10803607 TI - Development of high throughput screening assays using fluorescence polarization: nuclear receptor-ligand-binding and kinase/phosphatase assays. AB - Fluorescence polarization (FP) has been used to develop high throughput screening (HTS) assays for nuclear receptor-ligand displacement and kinase inhibition. FP is a solution-based, homogeneous technique requiring no immobilization or separation of reaction components. The FP-based estrogen receptor (ER) assay is based on the competition of fluorescein-labeled estradiol and estrogen-like compounds for binding to ER. These studies determined the Kd for this interaction to be 3 nM for ERalpha and 2 nM for ERbeta; IC50 values for 17beta-estradiol, tamoxifen, 4-OH-tamoxifen, and diethylstibestrol were determined to be 5.6, 189, 26, and 3.5 nM, respectively. In a screen of 50 lead compounds from a transcriptional activation screen, 21 compounds had IC50 values below 10 microM, with one having an almost 100-fold higher affinity for ERbeta over ERalpha. These data show that an FP-based competitive binding assay can be used to screen diverse compounds with a broad range of binding affinities for ERs. The FP-based protein-tyrosine kinase (PTK) assay uses fluorescein-labeled phosphopeptides bound to anti-phosphotyrosine antibodies. Phosphopeptides generated by a kinase compete for this binding. In c-Src kinase reactions, polarization decreased with time as reaction products displaced the fluorescein-labeled phosphopeptide from the anti-phosphotyrosine antibodies. The experimentally determined IC50 of AG 1478 was 400 pM, while Genistein did not inhibit the epidermal growth factor receptor at similar concentrations. Like the FP-based PTK assay, the protein kinase C (PKC) assay utilizes competition. PKC isoforms had different turnover rates for the peptide substrate. The IC50 for staurosporine was less than 10 nM for all PKC isoforms. Tyrosine phosphatase assays use direct binding rather than competition. Increasing concentrations of T-cell protein-tyrosine phosphatase (TC PTP) increased the rate of dephosphorylation. This change in polarization was dependent on TC PTP and was inhibited by 50 microM Na3VO4. The IC50 of Na3VO4 was 4 nM for TC PTP. These data demonstrate that a FP-based assay can detect kinase and phosphatase activity. Homogeneous, fluorescent techniques such as FP are now methods of choice for screening many types of drug targets. New HTS instrumentation and assay methods like these make FP a technology easily incorporated into HTS. PMID- 10803608 TI - Protein expression strategies for identification of novel target proteins. AB - Identification of new target proteins is a novel paradigm in drug discovery. A major bottleneck of this strategy is the rapid and simultaneous expression of proteins from differential gene expression to identify eligible candidates. By searching for a generic system enabling high throughput expression analysis and purification of unknown cDNAs, we evaluated the YEpFLAG-1 yeast expression system. We have selected cDNAs encoding model proteins (eukaryotic initiation factor-5A [eIF-5A] and Homo sapiens differentiation-dependent protein-A4) and cDNA encoding an unknown protein (UP-1) for overexpression in Saccharomyces cerevisiae using fusions with a peptide that changes its conformation in the presence of Ca2+ ions, the FLAG tag (Eastman Kodak, Rochester, NY). The cDNAs encoding unknown proteins originating from a directionally cloned cDNA library were expressed in all three possible reading frames. The expressed proteins were detected by an antibody directed against the FLAG tag and/or by antibodies against the model proteins. The alpha-leader sequence, encoding a yeast mating pheromone, upstream of the gene fusion site facilitates secretion into the culture supernatant. EIF-5A could be highly overexpressed and was secreted into the culture supernatant. In contrast, the Homo sapiens differentiation-dependent protein-A4 as well as the protein UP-1, whose cDNA did not match to any known gene, could not be detected in the culture supernatant. The expression product of the correct frame remained in the cells, whereas the FLAG-tagged proteins secreted into the supernatant were short, out-of-frame products. The presence of transmembrane domains or patches of hydrophobic amino acids may preclude secretion of these proteins into the culture supernatant. Subsequently, isolation and purification of the various proteins was accomplished by affinity chromatography or affinity extraction using magnetizable beads coated with the anti-FLAG monoclonal antibody. The purity of isolated proteins was in the range of 90%. In the case of unknown cDNAs, the expression product with the highest molecular mass was assumed to represent the correct reading frame. In summary, we consider the YEpFLAG-1 system to be a very efficient tool to overexpress and isolate recombinant proteins in yeast. The expression system enables high throughput production and purification of proteins under physiological conditions, and allows miniaturization into microtiter formats. PMID- 10803609 TI - CT evaluation of bladder trauma: a critical look. PMID- 10803610 TI - Clinical evaluation of wavelet-compressed digitized screen-film mammography. AB - RATIONALE AND OBJECTIVES: The authors compared diagnostic accuracy and callback rates with conventional screen-film mammograms and wavelet-compressed digitized images. MATERIALS AND METHODS: Sixty sets of mammograms (four views per case) were digitized at a spatial resolution of 100 microm. The images were wavelet compressed to a mean compression ratio of 8:1 and reviewed by three mammographers. Five regions were evaluated in each breast. Suspicion of malignancy was graded on a scale of 0% to 100%, and receiver operating characteristic (ROC) analysis was performed. Callback rates were calculated by using the American College of Radiology's Breast Imaging Reporting and Data System lexicon scale. RESULTS: The mean diagnostic accuracy with compressed and conventional images was 0.832 and 0.860, respectively. The upper 95% confidence bound for the difference in ROC areas was 0.061. The mean false-positive rate at a fixed sensitivity of 0.90 was 0.041 for compressed images and 0.059 for conventional images. The mean callback rates for normal, benign, and malignant regions were 0.023, 0.305, and 0.677, respectively, for compressed images and 0.036, 0.447, and 0.750, respectively, for conventional images. The upper 95% confidence bound for the (absolute) differences in callback rates was 0.012 for normal regions, 0.163 for benign regions, and 0.138 for malignant regions. CONCLUSION: Diagnostic accuracies were equivalent for both compressed and conventional images. The mean false-positive rate at fixed sensitivity was much better with the compressed images. However, the callback rates for malignant lesions were lower when the compressed images were used. PMID- 10803611 TI - Utility of routine trauma CT in the detection of bladder rupture. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine the frequency with which routine computed tomography (CT) fails to depict bladder rupture, the potential utility of delayed CT scans, and whether these findings might be useful in determining which patients may require subsequent cystography. MATERIALS AND METHODS: Cystograms and abdominal and pelvic CT scans of 54 patients with blunt trauma and in whom bladder rupture was clinically suspected were retrospectively reviewed. Blind readings of CT scans were performed by two genitourinary radiologists. Cystograms were used as the standard. RESULTS: Cystograms depicted bladder rupture in 10 patients. On CT scans, extravesical fluid was depicted in all three patients with intraperitoneal bladder rupture (although only a small amount of pelvic intraperitoneal fluid was present in two of these patients), in all seven patients with extraperitoneal bladder rupture, and in 32 of the 44 patients without bladder injury. Contrast material had been excreted into the bladder at the time of the initial or delayed CT in eight patients with bladder rupture; however, extravasation was identified in only four of the eight. In two of the four patients without extravasation, the bladder was distended at the time of CT. No bladder injuries were found in the 12 patients in whom pelvic fluid was not identified on CT scans. CONCLUSION: The absence of pelvic fluid on a trauma CT scan indicates that bladder rupture is unlikely. Even when a partially opacified bladder is passively distended, bladder injury may be present despite the absence of contrast material extravasation. PMID- 10803612 TI - Efficacy of targeted CT angiography in evaluation of intracranial internal carotid artery disease. AB - RATIONALE AND OBJECTIVES: This study evaluated the efficacy of targeted computed tomographic (CT) angiography in the diagnosis of intracranial internal carotid artery (ICA) disease and compared the results of routine and targeted CT angiography. MATERIALS AND METHODS: Fifty-four patients (24 male and 30 female patients aged 2 months to 87 years) were examined with CT angiography. Digital subtraction angiography (DSA) was performed in 42. CT angiograms were reconstructed with the maximum-intensity projection (MIP) algorithm. Targeted CT angiography was performed by individually reconstructing a single ICA territory. Each ICA was divided into four segments, and findings of routine MIP CT angiography, routine MIP plus targeted CT angiography, and DSA were reviewed independently by two neuroradiologists for vascular lesions involving each segment. Routine and targeted CT angiograms were also evaluated to determine how well both ICAs were visualized. RESULTS: Routine CT angiography was rated good or excellent for ICA visualization in 64% of cases, compared with 81% for targeted CT angiography (P = .0005). The overall agreement between routine CT angiography and DSA and between routine plus targeted CT angiography and DSA was 92% and 94%, respectively. There was no statistically significant difference between the percentages of vascular lesions detected with routine CT angiography alone and with routine plus targeted CT angiography. Both methods tended to show false positive findings of steno-occlusive disease, but targeted CT angiography showed details of aneurysms and stenotic lesions that were easily overlooked with routine CT angiography alone. CONCLUSION: Routine plus targeted CT angiography, while providing superior image quality, did not have much clinical effect; further assessment may be needed. PMID- 10803613 TI - Informing patients of diagnostic mammography results: mammographer's opinions. AB - PURPOSE: The authors' purpose was to determine mammographers' practices and attitudes regarding disclosing results of diagnostic mammograms to patients. MATERIALS AND METHODS: In 1995, the authors mailed a questionnaire to 500 members of the Society of Breast Imaging; 399 (80%) responded to the survey. RESULTS: Three-quarters of respondents stated that mammographers should disclose results to their patients, and approximately half were already doing so (52% for normal results, 51% for abnormal results). A sizable minority (25%) said that not telling patients was the best practice and identified several barriers to direct disclosure, including lack of time. Although bivariate analysis showed direct disclosure to be more common among female mammographers, the sex difference did not persist in multivariate analysis. In both bivariate and multivariate analyses, reading more than 100 mammograms per week and having a radiology practice in a university or academic setting were each strongly associated with direct disclosure. CONCLUSION: Implementation of the Mammography Quality Standards Reauthorization Act of 1998 may not require a major change in mammographers' current practice. It remains critical to establish systems that help radiologists disclose results and communicate with referring physicians efficiently and effectively. PMID- 10803614 TI - Components-of-variance models and multiple-bootstrap experiments: an alternative method for random-effects, receiver operating characteristic analysis. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to develop an alternative approach to random-effects, receiver operating characteristic analysis inspired by a general formulation of components-of-variance models. The alternative approach is a higher-order generalization of the Dorfman, Berbaum, and Metz (DBM) approach that yields additional information on the variance structure of the problem. MATERIALS AND METHODS: Six population experiments were designed to determine the six variance components in the DBM model. For practical problems, in which only a finite set of readers and patients are available, six analogous bootstrap experiments may be substituted for the population experiments to estimate the variance components. Monte Carlo simulations were performed on the population experiments, and those results were compared with the corresponding multiple-bootstrap estimates and those obtained with the DBM approach. Confidence intervals on the difference of ROC parameters for competing diagnostic modalities were estimated, and corresponding comparisons were made. RESULTS: For mean values, the agreement of present estimates of variance structures with population results was excellent and, when suitably weighted and mixed, similar to or closer than that with the DBM method. For many variance structures, the confidence intervals in this study for the difference in ROC area between modalities were comparable to those with the DBM method. When reader variability was large, however, mean confidence intervals from this study were tighter than those with the DBM method and closer to population results. CONCLUSION: The jackknife approach of DBM provides a linear approximation to receiver-operating characteristic statistics that are intrinsically nonlinear. The multiple bootstrap technique of this study, however, provides a more general, nonparametric, maximum-likelihood approach. It also yields estimates of the variance structure previously unavailable. PMID- 10803615 TI - Radiology resident performance reviews. PMID- 10803616 TI - Introduction to an academic career in radiology. PMID- 10803617 TI - Peptic ulcer disease: pathogenesis, radiologic features, and complications. PMID- 10803618 TI - Digital cameras in academic radiology practice. PMID- 10803619 TI - The National Institutes of Health fiscal year 2001 budget: implications for biomedical imaging research. PMID- 10803620 TI - Sensory recovery of innervated and non-innervated radial forearm free flaps: functional implications. AB - Findings reported in the literature on the sensation provided by intraorally applied innervated vs. non-innervated radial forearm free flaps differ. In an effort to understand these differences in sensory recovery, the authors carried out sensory evaluations in 12 patients who had undergone radial forearm free flaps. Seven patients had innervated flaps for defects of the tongue and floor of mouth; five had non-innervated flaps to various sites. Flap sensitivity to temperature, light touch, dull touch, and sharpness and two-point discrimination was assessed at the donor site and contralaterally, and at the recipient site and contralateral mirror-image oral mucosa. Patients subjectively rated post reconstruction sensation and provided quality of life (QOOL) data. The innervated flaps demonstrated better sensory recovery than the non-innervated flaps, although the latter did restore reasonable sensation. This paper describes the results, compares the study to other similar studies, and discusses various factors in the sensory recovery of both innervated and non-innervated intraoral radial forearm free flaps. The authors conclude that, although the trend in this study is toward improved function with the innervated flaps, these flaps do not appear to offer major intraoral functional advantage over the non-innervated flaps, which attain reasonably effective sensory recovery from neural ingrowth, if the lingual nerve is intact. PMID- 10803621 TI - Indications and limitations of angiography before free-flap transplantation to the distal lower leg after trauma: prospective study in 36 patients. AB - The necessity for routine angiography in evaluation of the vasculature of recipient legs prior to microsurgical free-tissue reconstruction still remains controversial. This prospective study was designed to determine the indications and limitations of angiography pertinent to this issue. The protocol consisted of palpation of dorsalis pedis and posterior tibial artery pulsation and lower-limb angiography prior to surgery, as well as intraoperative observation of the posterior tibial, anterior tibial, and peroneal arteries. Thirty-six patients were included who were treated from November, 1993 to December 1998. In five patients either the posterior tibial pulse (1), the dorsalis pedis pulse (3), or both pedal pulses, including the popliteal pulse (1) were not palpable preoperatively. These clinical findings correlated with the vascular lesion images on angiography. In two patients, pedal pulse palpation could not clearly be evaluated because of the injury. Among the 29 patients with both pedal pulses palpable, three patients angiographically presented an injury of the peroneal artery, and one patient a pseudoaneurysm of the anterior tibial artery. In none of the cases with at least one palpable pedal pulse (33), did preoperative angiography add relevant information which led to a plan change in the free-flap transfer. However, in two cases, severe scarring and fibrosis required an intraoperative change of the recipient vessel in one case, and a change of the anastomosis level and use of a vein graft, in the other case, although the angiography had demonstrated normal vascularity in both. The authors conclude that preoperative angiography is indicated only when both pedal pulses are not palpable, and that normal preoperative angiography does not guarantee the presence of vessels suitable for anastomosis. PMID- 10803622 TI - Nerve stimulating system for peripheral nerve tumor resection under the thoracoscope. AB - The authors report a newly devised nerve stimulator and its clinical application in thoracoscopic surgery. The stimulator is a monopolar system, that has a long extension between the electrodes and the handle. The stimulator is inserted into the chest cavity through a portal, and is used to distinguish motor nerves from the surrounding tissue and to determine whether there is nerve involvement with tumor. The tumor is then safely and rapidly resected by thoracoscopic technique and the major nerves are spared. This method was used in two clinical cases, with excellent results obtained, and with no major complications. PMID- 10803623 TI - Is it necessary to use the angular artery to feed the scapular tip when preparing a latissimus dorsi osteomyocutaneous flap?: case report. AB - This report demonstrates the possibility of elevation of the scapular tip with the latissimus dorsi muscle based on the thoracodorsal artery only, when an additional and substantial amount of bone is required for complex reconstruction. The patient was a 37-year-old man who developed an epidermoid carcinoma arising from the left maxillary sinus. After wide excision and radical resection of the tumor and the invaded structures, an osteomyocutaneous latissimus dorsi flap was prepared. With the muscle, the 12th rib was included in the flap to reconstruct the orbital floor and zygomatic arch, and the scapular tip was also elevated to reconstruct the hard palate. The skin island over the muscle was designed according to reconstructive requirements, including the buccal lining, nasal lateral wall lining, and coverage of the scapular tip at its new location to reconstruct the hard palate. All of these structures were successfully reconstructed with a single pedicle branch arising from the thoracodorsal artery. Postoperative early and late bone scans showed living bone at the zygomatic arch and hard palate. PMID- 10803624 TI - Management of alcohol withdrawal in microvascular head and neck reconstruction. AB - Alcohol use is a risk factor for head and neck cancer. One of the primary therapeutic modalities is surgical tumor ablation followed by immediate reconstruction. Such therapy places patients in a controlled environment, without alcohol, creating the risk of alcohol withdrawal syndrome. The authors attempted to identify the incidence of alcohol withdrawal among patients undergoing free flap reconstruction for head and neck cancer and were interested in the effect of alcohol withdrawal on the postoperative course of affected patients. In this retrospective review of 51 patients, six experienced alcohol withdrawal or delirium tremens. No difference in the rate of overall complications was seen between the patients who experienced withdrawal and those who did not, although patients who experienced withdrawal did have a statistically significant ratio of non-flap-related to flap-related complications. A review of the current management for alcohol withdrawal is included and places an emphasis on preoperative screening, close observation, rapid diagnosis, and immediate medical care. PMID- 10803625 TI - Inferior angle of the scapula as a vascularized bone graft: an anatomic study. AB - In reconstructing patients with massive midface losses, the authors required a vascularized bone graft that could be used to reconstruct the palate and orbital floor, provide vertical maxillary support, and provide soft tissue. The inferior angle of the scapula appears to be a promising source of vascularized bone stock, and until now, there has been no clear description of its vascular anatomy. The purpose of this study was to define the vascular anatomy of the inferior angle of the scapula. Thirteen fresh cadavers were studied. The subscapular artery was injected with Microfil (Flow Tec, Carver, MA) at its origin. Two branches of the subscapular artery were found to converge on the angle of the scapula: the descending osseous branch of the circumflex scapular, and the transverse branch of the thoracodorsal. The descending osseous branch of the circumflex scapular artery supplied the inferior angle of the scapula in 100 percent of cases. The transverse branch of the thoracodorsal artery supplied it in 76 percent of cases. The descending osseous branch of the circumflex scapular artery is the principal artery supplying the inferior angle of the scapula. PMID- 10803626 TI - James R. Learmonth: the first peripheral nerve surgeon. AB - The life and education of James R. Learmonth are reviewed. His experimental and clinical activities place him historically as the first surgeon to devote himself to peripheral nerve surgery. PMID- 10803627 TI - Long nerve allografts in sheep with Cyclosporin A immunosuppression. AB - Rodent studies of nerve allografts are limited by a relatively short length of graft segment. The authors attempted to establish an outbred sheep model that would allow the study of longer, more clinically relevant nerve gaps. Using outbred ewes, two 8-cm long radial sensory nerves were grafted into gaps (5 cm) in the median nerve. Sheep received an autograft and an allograft. Four sheep were immunosuppressed with Cyclosporin A (CsA) and four were controls. Blood CsA levels greater than 1000 microg/L were obtained. Systemic immunosuppression resulted in severe opportunistic infections, and the sheep were sacrificed between 35 and 47 days following surgery. Histologically, in the autografts and CsA-treated allografts, evidence of nerve regeneration was seen. Non immunosuppressed allografts were clearly rejected. While clear differences in the histology of experimental and control grafted nerve tissues were seen, the sheep allograft model presents considerable challenges due to immunosuppression-related infectious complications. PMID- 10803628 TI - Blockade of L-selectin attenuates reperfusion injury in a rat model. AB - Ischemia/reperfusion (I/R) injury appears to be a significant neutrophil dependent component and may be ameliorated by blocking leukocyte-endothelial adhesion. Using a rat extensor digitorum longus (EDL) muscle model, the present study tested the hypothesis that in vivo administration of the function-blocking monoclonal antibody (mAb) LAM1-116 which recognizes L-selectin, a cell-surface adhesion receptor, could decrease I/R injury. In 46 rats, one EDL served as a normal control and the opposite EDL underwent 3 hr of ischemia followed by 3 hr of reperfusion after pretreatment with LAM1-116 mAb, control IgG, or saline. Myeloperoxidase (MPO) activity showed only a two-fold increase from normal in LAM1-116-treated I/R EDL while a 27-fold increase occurred in the IgG2a and saline groups, with a statistically significant (p < 0.001) difference. A significantly (p < 0.05) lower wet weight ratio, improved fatigue contractile force, and less neutrophil infiltration were found in LAM1-116-treated EDL, when compared to those in control IgG- or saline-treated EDL. The results indicate that blockade of L-selectin by LAM1-116 mAb can effectively reduce neutrophil infiltration in reperfused skeletal muscle, thereby decreasing tissue edema and improving muscle fatigue contractile force. These findings may be important in understanding I/R injury. PMID- 10803629 TI - Composite tissue allotransplantation: a comprehensive review of the literature- part III. PMID- 10803630 TI - The natural history of hepatitis C viral infection. AB - Although early data suggested that chronic hepatitis C virus infection carried little risk, studies with longer duration of infection have reported concerning results. Of patients with acute infection, approximately 80% will develop chronic infection. The greatest risk of morbidity comes with cirrhosis and the resulting increased risk of hepatocellular carcinoma. The true risk of progression to cirrhosis, however, has emerged as an area of controversy. Both host and viral factors seem to impact susceptibility to chronic infection, cirrhosis, and hepatocellular carcinoma. Hepatitis C virus has become the most common indication for liver transplantation, but the infection routinely recurs and may have a more aggressive course after transplantation. Given that current treatment options for hepatitis C virus infection are clearly not optimal, informed decisions regarding treatment require an in depth understanding of the natural history. PMID- 10803631 TI - Extrahepatic manifestations of chronic hepatitis C. AB - A by-product of increasing experience with patients infected with the hepatitis C virus is the awareness of a variety of extrahepatic syndromes that seem to be associated with HCV infection. Recent investigations into the relationship between the hepatitis C virus and human cells, particularly lymphocytes, have resulted in possible pathophysiological interactions that may begin to explain some of the extrahepatic manifestations of hepatitis C virus infection. In this review, we will discuss some of the potential interactions from both pathophysiological and clinical viewpoints. PMID- 10803632 TI - Hepatic fibrogenesis and hepatitis C. AB - Hepatitis C virus (HCV) infection is currently the most common cause of fibrosing liver disease and represents a major clinical challenge. In patients with HCV infection, inflammation and injury lead to fibrosis and cirrhosis in a significant proportion of patients; cirrhosis in turn has multiple clinical sequelae. Therefore, understanding the pathological basis of fibrogenesis in hepatitis C infection is critical. This review will highlight fundamental issues underlying the fibrogenic response to injury and in addition will focus on potential points of intervention. PMID- 10803633 TI - Which patients with hepatitis C virus should be treated? AB - Since the National Institutes of Health (NIH) Consensus Conference in 1997, our understanding of the natural history of hepatitis C (HCV) infection and our ability to treat patients has improved. Thus, a large number of clinical studies, confounding terminology, and a growing dilemma in targeting particular populations for treatment who have HCV infection, will continue to be at the forefront of clinical research and treatment. In this report, we examine which HCV-infected populations of patients should be treated. Beginning with treatment guidelines from the NIH Consensus Conference, and a brief overview of the terminology used in the HCV literature, we subsequently review data regarding treatment outcomes based on HCV viral load, genotype, and various epidemiological factors. Similarly, more challenging treatment strategies are discussed for patients with HCV infection, including those with ongoing psychiatric disorders, patients who are coinfected with the human immunodeficiency virus and HCV, and those patients with normal serum transaminases. Finally, a review and guidelines about other HCV treatment dilemmas, including patients with chronic renal failure on hemodialysis, patients who have undergone renal transplantation, and treatment of patients acutely exposed to HCV are also addressed. PMID- 10803634 TI - Hepatitis C virus and liver transplantation. AB - Hepatitis C virus (HCV) infection is the most common indication for liver transplantation in North America and Europe. While hepatitis C recurrence is common after transplantation, 5-year graft and patient survival in HCV-infected patients are similar to those of patients transplanted for other chronic liver diseases. With longer periods of follow-up, the proportion of patients with fibrosis or cirrhosis increases and graft loss does occur because of recurrent disease. Both viral and host factors have been linked to risk of disease progression. Specific therapies to eradicate infection or slow down disease progression are under study, and the most promising results to date have been obtained with combined interferon and ribavirin. PMID- 10803635 TI - Chronic hepatitis C: retreatment of relapsers. An evidence-based approach. AB - Post-treatment relapse remains a major issue in the long-term management of chronic hepatitis C. Many studies have been conducted to identify the ideal therapy that would increase the cost-effectiveness of retreatment in the individual patient. Although the conclusions of two consensus conferences for the retreatment of relapse of chronic hepatitis C have been published recently, several important issues still remain unanswered. We reviewed the available data by an evidence-based approach and conclude the following: (1) patients should be retreated with a combination of interferon (IFN) and ribavirin for 6 months if there are no contraindications to ribavirin; (2) the excellent tolerability and the lesser expense of retreatment with IFN monotherapy makes it a low-cost option for patients who have transiently cleared HCV-RNA during the first IFN course, and a primary indication for those who are contraindications to ribavirin or are likely to experience adverse events under ribavirin; (3) relapsers retreated with monotherapy must receive a high dose of IFN; and (4) patients with cirrhosis should not be retreated with IFN alone. More data, particularly on the long-term course of patients retreated with combination therapy, are needed before setting guidelines for retreatment of relapsers. PMID- 10803636 TI - Brain imaging studies of cocaine abuse: implications for medication development. AB - Contemporary in vivo brain imaging techniques confer the ability to assess brain function and structure noninvasively, and thereby can yield information to help guide the development of new treatments for substance abuse. The advantages and limitations of the major imaging modalities (positron emission tomography [PET], single photon emission computed tomography [SPECT], structural and functional magnetic resonance imaging [MRI, fMRI, respectively]) are discussed with respect to their applicability to research on cocaine abuse. The effects of acute administration of cocaine have been studied using PET and fMRI, with PET manifesting decreases in cerebral glucose metabolism and blood flow, and fMRI revealing regional effects that are correlated temporally with subjective responses. In addition, studies of drug abusers, abstinent from cocaine for various lengths of time, have revealed persistent differences in brain function and structure, especially in the frontal cortex, when compared with parameters in the brains of subjects who do not use illicit drugs of abuse. PET studies also have revealed abnormalities in markers for dopaminergic and opioid systems during withdrawal from cocaine. Moreover, studies of cue-elicited craving for cocaine demonstrate a connection between the response to drug-related stimuli and neural elements of cognition and emotion. The future directions of in vivo brain imaging to identify functional and structural alterations in the brains of cocaine abusers are discussed in relation to the development of medications to treat cocaine dependence. PMID- 10803637 TI - Behavioral effects of cannabinoid agents in animals. AB - Two subtypes of cannabinoid receptors have been identified to date, the CB1 receptor, essentially located in the CNS, but also in peripheral tissues, and the CB2 receptor, found only at the periphery. The identification of delta9 tetrahydrocannabinol (delta9-THC) as the major active component of marijuana (Cannabis sativa), the recent emergence of potent synthetic ligands and the identification of anandamide and sn-2 arachidonylglycerol as putative endogenous ligands for cannabinoid receptors in the brain, have contributed to advancing cannabinoid pharmacology and approaching the neurobiological mechanisms involved in physiological and behavioral effects of cannabinoids. Most of the agonists exhibit nonselective affinity for CB1/CB2 receptors, and delta9-THC and anandamide probably act as partial agonists. Some recently synthesized molecules are highly selective for CB2 receptors, whereas selective agonists for the CB1 receptors are not yet available. A small number of antagonists exist that display a high selectivity for either CB1 or CB2 receptors. Cannabinomimetics produce complex pharmacological and behavioral effects that probably involve numerous neuronal substrates. Interactions with dopamine, acetylcholine, opiate, and GABAergic systems have been demonstrated in several brain structures. In animals, cannabinoid agonists such as delta9-THC, WIN 55,212-2, and CP 55,940 produce a characteristic combination of four symptoms, hypothermia, analgesia, hypoactivity, and catalepsy. They are reversed by the selective CB1 receptor antagonist, SR 141716, providing good evidence for the involvement of CB1-related mechanisms. Anandamide exhibits several differences, compared with other agonists. In particular, hypothermia, analgesia, and catalepsy induced by this endogenous ligand are not reversed by SR 141716. Cannabinoid-related processes seem also involved in cognition, memory, anxiety, control of appetite, emesis, inflammatory, and immune responses. Agonists may induce biphasic effects, for example, hyperactivity at low doses and severe motor deficits at larger doses. Intriguingly, although cannabis is widely used as recreational drug in humans, only a few studies revealed an appetitive potential of cannabimimetics in animals, and evidence for aversive effects of delta9-THC, WIN 55,212-2, and CP 55,940 is more readily obtained in a variety of tests. The selective blockade of CB1 receptors by SR 141716 impaired the perception of the appetitive value of positive reinforcers (food, cocaine, morphine) and reduced the motivation for sucrose, beer and alcohol consumption, indicating that positive incentive and/or motivational processes could be under a permissive control of CB1-related mechanisms. There is little evidence that cannabinoid systems are activated under basal conditions. However, by using SR 141716 as a tool, a tonic involvement of a CB1-mediated cannabinoid link has been demonstrated, notably in animals suffering from chronic pain, faced with anxiogenic stimuli or highly motivational reinforcers. Some effects of SR 141716 also suggest that CB1-related mechanisms exert a tonic control on cognitive processes. Extensive basic research is still needed to elucidate the roles of cannabinoid systems, both in the brain and at the periphery, in normal physiology and in diseases. Additional compounds, such as selective CB1 receptor agonists, ligands that do not cross the blood brain barrier, drugs interfering with synthesis, degradation or uptake of endogenous ligand(s) of CB receptors, are especially needed to understand when and how cannabinoid systems are activated. In turn, new therapeutic strategies would likely to emerge. PMID- 10803638 TI - Multiple chemical sensitivity: potential role for neural sensitization. AB - An emerging issue in environmental health is the phenomenon of multiple chemical sensitivity (MCS). Multiple chemical sensitivity is a controversial disorder characterized by multiorgan symptoms in response to low-level chemical exposures that are considered safe for the general population. The onset of MCS is often attributed to prior repeated chemical exposures in the home and/or workplace, and, once initiated, symptoms are triggered by extremely low levels of many chemicals/foods. No single case definition exists for MCS due to several issues that call into question its validity as a distinct illness induced by prior chemical exposure. Hypotheses regarding the etiological basis for MCS range from direct toxicological effects of chemicals to the notion that MCS is purely a psychological "belief system". One leading hypothesis suggests that MCS represents a neural sensitization phenomenon, wherein susceptible individuals demonstrate extreme sensitivity to chemicals and odor intolerance due to central nervous system (CNS) sensitization processes. The recent development of an animal model for MCS provides some support for the sensitization hypothesis and may offer evidence for behavioral changes observed in at least a subset of those reporting MCS. PMID- 10803639 TI - Expression of cyclooxygenase-2 in transitional cell carcinoma of the urinary bladder in dogs. AB - OBJECTIVE: To evaluate expression of cyclooxygenase (COX)-1 and COX-2 in the urinary bladder epithelium of clinically normal dogs and in transitional cell carcinoma cells of dogs. ANIMALS: 21 dogs with transitional cell carcinoma of the urinary bladder and 8 dogs with clinically normal urinary bladders. PROCEDURE: COX-1 and COX-2 were evaluated by use of isoform-specific antibodies with standard immunohistochemical methods. RESULT: COX-1, but not COX-2, was constitutively expressed in normal urinary bladder epithelium; however, COX-2 was expressed in neoplastic epithelium in primary tumors and in metastatic lesions of all 21 dogs and in new proliferating blood vessels in 3 dogs. Also, COX-1 was expressed in the neoplastic cells. CONCLUSIONS AND CLINICAL RELEVANCE: Lack of expression of COX-2 in normal bladder epithelium and its substantial expression in transitional cell carcinoma cells suggest that this isoform may be involved in tumor cell growth. Inhibition of COX-2 is a likely mechanism of the antineoplastic effects of non steroidal antiinflammatory drugs. PMID- 10803640 TI - Safety of moxidectin in avermectin-sensitive collies. AB - OBJECTIVE: To evaluate the safety of moxidectin administration at doses of 30, 60, and 90 microg/kg of body weight (10, 20, and 30 times the manufacturer's recommended dose) in avermectin-sensitive Collies. ANIMALS: 24 Collies. PROCEDURE: Collies with mild to severe reactions to ivermectin challenge (120 mg/kg; 20 times the recommended dose for heartworm prevention) were used. Six replicates of 4 dogs each were formed on the basis of body weight and severity of reaction to ivermectin test dose. Within replicates, each dog was randomly allocated to treatment with oral administration of 30, 60, or 90 microg of moxidectin/kg or was given a comparable volume of placebo tablet formulation. Dogs were observed hourly for the first 8 hours and twice daily thereafter for 1 month for signs of toxicosis. RESULTS: Signs of toxicosis were not observed in any control group dog throughout the treatment observation period. Likewise, signs of toxicosis were not observed in any dog receiving moxidectin at 30, 60, or 90 microg/kg. CONCLUSIONS AND CLINICAL RELEVANCE: The moxidectin formulation used in the study reported here appears to have a wider margin of safety than ivermectin or milbemycin in avermectin-sensitive Collies. PMID- 10803641 TI - Evaluation of a method for experimental induction of osteoarthritis of the hip joints in dogs. AB - OBJECTIVE: To evaluate a method for experimental induction of osteoarthritis in the hip joints of dogs. ANIMALS: 12 mixed-breed dogs. PROCEDURE: A unilateral triple pelvic osteotomy was performed. In 6 dogs, the iliac osteotomy was repaired with 45 degrees of internal rotation, reducing coverage of the femoral head by the acetabulum. In the other 6 dogs, the fragments were repaired in anatomic alignment. Radiography, force plate evaluations, and subjective lameness evaluations were performed before and after surgery. Dogs were euthanatized 7 months after surgery, and samples of cartilage and joint capsule were examined histologically. RESULTS: Subjective lameness scores, radiographic appearance of the hip joints, and Norberg angles were not significantly different between groups; however, force plate evaluations did reveal significant differences in vertical ground reaction forces. Femoral head coverage was significantly decreased with rotation of the acetabulum. Mild inflammatory changes were discernible in the joint capsule and articular cartilage of some dogs in both groups. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that 45 degrees internal rotation of the acetabulum does not consistently induce biologically important osteoarthritic changes in the hip joints of dogs. PMID- 10803642 TI - Role of inflammatory mediators in priming, activation, and deformability of bovine neutrophils. AB - OBJECTIVE: To determine the capacity of inflammatory mediators tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), platelet-activating factor (PAF), lipopolysaccharide (LPS), and leukotoxin to prime, activate, or alter deformability of adult bovine neutrophils. SAMPLE POPULATION: Blood collected from 5 healthy adult Holstein cows. PROCEDURE: Isolated neutrophils or whole blood was incubated with TNF-alpha, IL-8, PAF, LPS, or leukotoxin, and neutrophil chemiluminescence, degranulation, deformability, shape change, CD11b expression, and size distribution was measured. RESULTS: Incubation with TNF-alpha, IL-8, PAF, and LPS primed neutrophils for oxygen radical release but caused minimal oxygen radical release by themselves. None of the inflammatory mediators induced degranulation. Incubation with TNF-alpha and PAF resulted in a decrease in neutrophil deformability and induced shape change in neutrophils. Incubation with PAF consistently resulted in an increase in neutrophil size as measured by use of flow cytometry. Only IL-8 caused an increase in expression of CD11b by neutrophils. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammatory mediators tested had minimal effects on neutrophil oxygen radical production or degranulation but did prime neutrophils for oxygen radical production. Incubation with PAF and TNF alpha caused a decrease in neutrophil deformability and altered neutrophil shape and size. Results of our study indicate that PAF- and TNF-alpha-induced changes in neutrophil deformability and size may cause integrin- and selectin-independent trapping of neutrophils in the lungs of cattle with pneumonic pasteurellosis. PMID- 10803643 TI - Effects of polysulfated glycosaminoglycan and hyaluronan on prostaglandin E2 production by cultured equine synoviocytes. AB - OBJECTIVE: To investigate effects of the anti-arthritic agents hyaluronan and polysulfated glycosaminoglycan (PSGAG) on inflammatory metabolism in cultured equine synoviocytes. SAMPLE POPULATION: Synoviocytes cultured from samples obtained from the metacarpophalangeal joints of 4 horses. PROCEDURE: Equine synoviocytes were grown in monolayer culture. Synoviocytes were stimulated with lipopolysaccharide (LPS) and simultaneously treated with various concentrations of hyaluronan or PSGAG for 48 hours. Three hyaluronan preparations were compared. Prostaglandin E2 (PGE2) concentrations in culture medium were measured, using radioimmunoassay. RESULTS: The highest concentrations of hyaluronan and PSGAG tested inhibited PGE2 production. CONCLUSIONS AND CLINICAL RELEVANCE: Clinically achievable concentrations of hyaluronan and PSGAG inhibited PGE2 synthesis by cultured equine synoviocytes. This anti-inflammatory action may be a mechanism through which these agents exert anti-arthritic effects. The effect was obtained at concentrations that can be achieved by use of intra-articular, but not systemic, administration of hyaluronan or PSGAG. PMID- 10803644 TI - Heart rate variability in Doberman Pinschers with and without echocardiographic evidence of dilated cardiomyopathy. AB - OBJECTIVE: To characterize the salient variables of the time-domain analysis of heart rate variability (HRV) in clinically normal Doberman Pinschers and to compare those variables with those of Doberman Pinschers with cardiomyopathy and mild to moderate myocardial failure. ANIMALS: 46 Doberman Pinschers. PROCEDURE: HRV was analyzed in the time-domain from 24-hour Holter recordings obtained from 28 Doberman Pinschers with normal echocardiograms and 18 Doberman Pinschers with echocardiograms consistent with mild to moderate myocardial failure. RESULTS: Significant differences in HRV variables between the 2 groups of dogs were not detected. The HRV was greater during the nighttime (12 AM to 6 AM), compared with the 24-hour day and an 18-hour (6 AM to 12 AM) period. CONCLUSIONS AND CLINICAL RELEVANCE: HRV of dogs with mild to moderate myocardial failure was not different from that of clinically normal dogs, because there were no disturbances of autonomic balance, baroreceptor function, and other factors that influence HRV in the dogs with cardiomyopathy, or the sensitivity of time-domain analysis was overwhelmed by normal sinus arrhythmia. The techniques now used to study HRV have important limitations, especially in dogs, and better noninvasive tests of autonomic function are needed. PMID- 10803645 TI - Oxidant stress in sled dogs subjected to repetitive endurance exercise. AB - OBJECTIVE: To determine whether repetitive endurance exercise in sled dogs was associated with substantial lipid peroxidation, decreases in antioxidant capacity of the serum, and skeletal muscle damage. ANIMALS: 24 lightly trained sled dogs. PROCEDURE: 16 dogs completed a 58-km run on each of 3 consecutive days; the other 8 dogs (control) did not exercise during the study. Blood samples were collected before the first exercise run and after the first and third exercise runs. Plasma isoprostane and serum vitamin E concentrations, total antioxidant status of plasma, and serum creatine kinase activity were measured. RESULTS: Plasma isoprostane concentrations in dogs in the exercise group were significantly increased after the first exercise run and further significantly increased after the third exercise run. Serum vitamin E concentration was significantly decreased after the first exercise run in dogs in the exercise group, and this change persisted after the third exercise run. There was a significant linear relationship between plasma isoprostane concentration and the logarithm of serum creatine kinase activity (adjusted ? = 0.84). CONCLUSIONS AND CLINICAL RELEVANCE: Results demonstrate that repetitive endurance exercise in dogs is associated with lipid peroxidation and a reduction in plasma antioxidant concentrations. We interpret these results as indicating that the antioxidant mechanisms of minimally trained dogs may, in some instances, be inadequate to meet the antioxidant requirements of repetitive endurance exercise. PMID- 10803646 TI - Evaluation of intestinal permeability and gluten sensitivity in Soft-Coated Wheaten Terriers with familial protein-losing enteropathy, protein-losing nephropathy, or both. AB - OBJECTIVE: To evaluate intestinal permeability and gluten sensitivity in a family of Soft-Coated Wheaten Terriers (SCWT) affected with protein-losing enteropathy (PLE), protein-losing nephropathy (PLN), or both. ANIMALS: 6 affected adult dogs. PROCEDURE: Intestinal biopsy specimens, urine protein-to-creatinine ratio, serum concentrations of albumin and globulin, and concentration of alpha1-protease inhibitor in feces were evaluated before, during, and 13 weeks after daily administration of 10 g of gluten for 7 weeks. Eosinophils and lymphocytes plasmacytes were enumerated in intestinal biopsy specimens. Intestinal permeability was evaluated before and during the sixth week of gluten administration via cellobiose-mannitol and chromium-EDTA absorption tests. RESULTS: Serum globulin concentration decreased significantly after prolonged administration of gluten. Although not significant, there was an increase in lymphocytes-plasmacytes and a decrease in eosinophils in intestinal biopsy specimens. Furthermore, these counts were greater than those reported for clinically normal dogs. Gluten administration did not increase intestinal permeability. CONCLUSIONS AND CLINICAL RELEVANCE: Daily administration of gluten was associated with a significant decrease in serum globulin concentration in SCWT affected with PLE or PLN, but other variables remained unchanged. Although enhanced wheat-gluten sensitivity may be one factor involved in the pathogenesis of PLE or PLN in SCWT, this syndrome does not appear to be the result of a specific sensitivity to gluten. PMID- 10803647 TI - Effects of tilmicosin treatment on Pasteurella haemolytica organisms in nasal secretion specimens of calves with respiratory tract disease. AB - OBJECTIVE: To determine the effect of tilmicosin treatment on number of Pasteurella haemolytica (PH) organisms in nasal secretion specimens of calves with respiratory tract disease. ANIMALS: 206 British mixed-breed beef calves, 2 to 5 months old. PROCEDURE: In 2 separate studies of outbreaks, calves (study 1, n = 101; study 2, n = 105) that developed respiratory tract disease after transport to a feedlot were treated with tilmicosin. Nasal secretion specimens were examined for PH organisms to determine the status of colonization. RESULTS: In both studies, PH serotypes A1 and A6 were isolated. In study 1, tilmicosin treatment eliminated or markedly reduced the number of PH organisms in calves on days 1, 4, and 5 after treatment. In study 2, tilmicosin treatment eliminated PH organisms in calves on days 1, 2, 5, and 6 after treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Overall, tilmicosin treatment increased the number of culture positive calves that became culture-negative and decreased the number of culture negative calves that became culture-positive for up to 6 days after treatment. Tilmicosin treatment decreased the number of PH organisms in nasal secretion specimens, which indicated that fewer PH organisms were available to infect the lungs or to infect other calves. By reducing colonization, prophylactic use of tilmicosin before transport or at the time of arrival at a feedlot is likely to reduce the incidence of acute respiratory tract disease in calves for the initial several days after arrival, which is the period when they are most susceptible to infectious organisms. PMID- 10803648 TI - Nitric oxide metabolite production in the cranial cruciate ligament, synovial membrane, and articular cartilage of dogs with cranial cruciate ligament rupture. AB - OBJECTIVE: To measure concentrations of nitric oxide metabolites (nitrite-nitrate [NOt]) in cartilage, synovial membrane, and cranial cruciate ligament (CCL) in dogs and evaluate associations with osteoarthritis in dogs with CCL rupture. ANIMALS: 46 dogs with CCL rupture and 54 control dogs without joint disease. PROCEDURE: Tissue specimens for histologic examination and explant culture were harvested during surgery in the CCL group or immediately after euthanasia in the control group; NOt concentrations were measured in supernatant of explant cultures and compared among dogs with various degrees of osteoarthritis and between dogs with and without CCL rupture. RESULTS: Osteoarthritic cartilage had significantly higher NOt concentration (1,171.6 nmol/g) than did healthy cartilage (491.0 nmol/g); NOt concentration was associated with severity of macroscopic and microscopic lesions. Synovial membrane NOt concentration did not differ between dogs with and without CCL rupture. Ruptured CCL produced less NOt than did intact ligaments. In control dogs, NOt concentrations were similar for intact ligaments (568.1 nmol/g) and articular cartilage (491.0 nmol/g). Synthesis of NOt was inhibited substantially by coincubation with inhibitors. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that NOt in canine joint tissues originates from the inducible nitric oxide synthase pathway. Nitric oxide metabolite production in cartilage was greater in dogs with osteoarthritis than in healthy dogs and was associated with lesion severity, suggesting that nitric oxide inhibitors may be considered as a treatment for osteoarthritis. The CCL produces substantial concentrations of NOt; the importance of this finding is unknown. PMID- 10803649 TI - Effects of phenylbutazone on bone activity and formation in horses. AB - OBJECTIVE: To determine the effects of phenylbutazone (PBZ) on bone activity and bone formation in horses. ANIMALS: 12 healthy 1- to 2-year-old horses. PROCEDURES: Biopsy was performed to obtain unicortical bone specimens from 1 tibia on day 0 and from the contralateral tibia on day 14. Fluorochromic markers were administered IV 2 days prior to and on days 0, 10, 15, and 25 after biopsy was performed. Six horses received PBZ (4.4 mg/kg of body weight, PO, q 12 h) and 6 horses were used as controls. All horses were euthanatized on day 30 and tissues from biopsy sites, with adjacent cortical bone, were collected. Osteonal density and activity, mineral apposition rate (MAR), and percentage of mineralized tissue filling the biopsy-induced defects in cortical bone were assessed. Serum samples from all horses were analyzed for bone-specific alkaline phosphatase activity and concentration of PBZ. RESULTS: MAR was significantly decreased in horses treated with PBZ. Regional acceleratory phenomenon was observed in cortical bone in both groups but was significantly decreased in horses treated with PBZ. Osteonal activity was similar at all time points in all horses. In control horses, percentage of mineralized tissue filling the cortical defects was significantly greater in defects present for 30 days, compared with defects present for 14 days. Differences in percentage of mineralized tissue were not detected in horses treated with PBZ. CONCLUSIONS AND CLINICAL RELEVANCE: PBZ decreased MAR in cortical bone and appeared to decrease healing rate of cortical defects in horses. PMID- 10803650 TI - Evaluation of intratesticular injection of glycerol for nonsurgical sterilization of dogs. AB - OBJECTIVE: To determine efficacy of intratesticular injection of glycerol as a means to sterilize dogs. ANIMALS: Six 2- to 4-year-old sexually intact male dogs. PROCEDURE: Testicular volume, serum testosterone concentration, and number of sperm in ejaculate were determined before and at weekly intervals for 12 weeks following intratesticular injection of a 70% glycerol solution (1 ml/testis). Testicular tissue specimens for histologic evaluation were obtained 4, 8, and 12 weeks following injection. RESULTS: Number of sperm in ejaculate following glycerol injection was not significantly different from number before injection. Compared with preinjection values, serum testosterone concentrations were significantly less 5, 9, and 10 weeks following injection, although libido and ejaculate characteristics were unchanged. Testicular swelling was evident 24 to 48 hours following injection, and testicular volume decreased starting on week 5 following treatment. Seminiferous tubules and evidence of normal spermatogenesis were detected in testicular biopsy specimens obtained from all dogs 12 weeks following injection. CONCLUSION AND CLINICAL RELEVANCE: Intratesticular injection of a 70% glycerol solution did not result in azoospermia and sterility in dogs. PMID- 10803651 TI - Complex segregation analysis of deafness in Dalmatians. AB - OBJECTIVE: To use pedigree analysis to evaluate the feasibility of a major locus model for deafness in Dalmatians. ANIMALS: 605 purebred Dalmatians from 42 families. PROCEDURE: Hearing loss was evaluated through the brainstem auditory evoked response. Dogs were classified into mutually exclusive categories: normal hearing, unilaterally deaf, or bilaterally deaf. Information was collected on sex, coat color, presence or absence of a color patch at birth, and eye color. Statistical analyses were performed by use of regressive logistic models designed for complex segregation analysis. Genetic correlations among eye color, deafness, and color patch were estimated. RESULTS: Prevalence of hearing loss was 11% for dogs classified as unilaterally deaf and 5% for dogs that were bilaterally deaf. Complex segregation analysis detected statistical evidence of a single allele with an expected frequency of 0.21 that had an effect on the prevalence of deafness. Results of analyses suggested that this locus cannot completely explain the inheritance and incidence of deafness in Dalmatians. Genetic correlation estimates among deafness, eye color, and color patch revealed strong interrelationships among these characteristics. CONCLUSIONS AND CLINICAL RELEVANCE: To reduce the incidence of hearing loss in Dalmatians, unilaterally deaf, blue-eyed dogs should not be considered as potential parents. PMID- 10803652 TI - Canine antibody response to Blastomyces dermatitidis WI-1 antigen. AB - OBJECTIVE: To assess whether dogs with blastomycosis produce antibodies against the WI-1 and A-antigens of Blastomyces dermatitidis and whether the antibodies are useful in serodiagnosis. SAMPLE POPULATION: 359 serum samples obtained from 245 dogs. PROCEDURE: 233 samples from 122 dogs with blastomycosis, and 1 sample each from 24 dogs with suspected blastomycosis, 51 control dogs without infection, and 48 healthy dogs from an enzootic region were obtained. Antibodies against WI-1 antigen were detected by radioimmunoassay (RIA). Serum samples were tested in parallel for antibodies against the A-antigen of B dermatitidis by commercial agar-gel immunodiffusion (AGID) in a reference laboratory. RESULTS: Antibodies were detected in 92% of infected dogs by RIA and in 41 % by AGID. For 29 serum samples that were obtained 11 to 1,545 days after diagnosis, antibodies were detected in 92% of samples by RIA and 7% by AGID. For 93 serial serum samples from 29 dogs with blastomycosis, the mean anti-WI-1 titer was 1:18,761 at the time of diagnosis, and decreased to a mean of 1:1,338 by 210 days after treatment was initiated. Of 24 dogs with suspected infection, antibodies were detected in 67% by RIA and 33% by AGID. Control dogs without blastomycosis had no detectable antibodies in either assay. Thus, sensitivity was 92% for RIA and 41 % for AGID, and specificity was 100% for both tests. CONCLUSIONS AND CLINICAL RELEVANCE: Anti-WI-1 antibodies are readily detected by RIA in dogs with blastomycosis. Titers become high, decline during treatment, and persist for months. Anti-A antibodies are sometimes detected with AGID, but these decrease quickly. PMID- 10803653 TI - Influence of dietary protein and lipid on weight loss in obese ovariohysterectomized cats. AB - OBJECTIVE: To determine effects of dietary lipid and protein on development of hepatic lipidosis (HL) and on physical and biochemical indices following rapid weight loss in cats. ANIMALS: 24 ovariohysterectomized cats. PROCEDURE: Cats were fed a high energy diet until they gained 30% of their ideal body weight and then randomly assigned to receive 1 of 4 weight-reduction diets (6 cats/diet) at 25% of maintenance energy requirements per day. Diets contained a low or high quality protein source and a lipid source deficient or sufficient in long chain essential fatty acids (LCEFA). Serum and plasma samples and liver biopsy specimens were obtained for biochemical analyses and determination of hepatic lipid content before and after weight gain and during and after weight loss. RESULTS: Irrespective of weight-reduction diet fed, all cats lost weight at a comparable rate (4.51 to 5.00 g/d/kg of obese body weight). Three cats developed hepatic lipidosis. Significant changes in plasma insulin, cholesterol, triglyceride, and serum glucose concentrations were detected after weight gain and weight loss in all diet groups, but values for these variables did not differ among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Cats can lose 25 to 30% of their obese body weight over 7 to 9 weeks without developing overt clinical signs of HL, provided that weight-reduction diets are highly palatable, contain a high quality protein, have a source of LCEFA, and are fortified with vitamins and microminerals. However, rapid weight loss may increase risk factors associated with development of diabetes mellitus. PMID- 10803654 TI - Effect of dietary protein quality and fatty acid composition on plasma lipoprotein concentrations and hepatic triglyceride fatty acid synthesis in obese cats undergoing rapid weight loss. AB - OBJECTIVE: To determine effects of dietary lipid and protein on plasma lipoprotein and free fatty acid concentrations and hepatic fatty acid synthesis during weight gain and rapid weight loss in cats. ANIMALS: 24 ovariohysterectomized cats. PROCEDURE: Cats were fed a high energy diet until they gained 30% of their ideal body weight and then randomly assigned to receive 1 of 4 weight reduction diets (6 cats/diet) at 25% of maintenance energy requirements. Diets contained a low or high quality protein source and a lipid source deficient or sufficient in long chain essential fatty acids. Plasma samples and liver biopsy specimens were obtained before and after weight gain and during and after weight loss for determination of free fatty acid, triglyceride, and lipoprotein concentrations. Synthesis of these substances was measured by use of isotope enrichment. RESULTS: Plasma total cholesterol concentration and concentration of lipoprotein fractions increased after weight gain, compared with baseline values. Weight loss resulted in a significant decrease in concentrations of all lipoprotein fractions except high density lipoprotein. High density lipoprotein concentration was significantly greater in cats fed diets containing an oil blend, compared with cats fed diets containing corn oil. Fatty acid synthesis after weight loss was below the detection limit of the measurement technique. CONCLUSIONS AND CLINICAL RELEVANCE: In cats undergoing rapid weight loss there is neither increased triglyceride synthesis nor decreased transport of very low density lipoproteins from the liver, suggesting that their involvement in the development of hepatic lipidosis may be minimal. PMID- 10803655 TI - Pharmacokinetics of fenbendazole following intravenous and oral administration to pigs. AB - OBJECTIVE: To determine pharmacokinetics and metabolic patterns of fenbendazole after IV and oral administration to pigs. ANIMALS: 4 mixed-breed female pigs weighing 32 to 45 kg. PROCEDURE: Fenbendazole was administered IV at a dose of 1 mg/kg. One week later, it was administered orally at a dose of 5 mg/kg. Blood samples were collected for up to 72 hours after administration, and plasma concentrations of fenbendazole, oxfendazole, and fenbendazole sulfone were determined by use of high-pressure liquid chromatography. Plasma pharmacokinetics were determined by use of noncompartmental methods. RESULTS: Body clearance of fenbendazole after IV administration was 1.36 L/h/kg, volume of distribution at steady state was 3.35 L/kg, and mean residence time was 2.63 hours. After oral administration, peak plasma concentration of fenbendazole was 0.07 microg/ml, time to peak plasma concentration was 3.75 hours, and mean residence time was 15.15 hours. Bioavailability of fenbendazole was 27.1%. Oxfendazole was the major plasma metabolite, accounting for two-thirds of the total area under the plasma concentration versus time curve after IV and oral administration. Fenbendazole accounted for 8.4% of the total AUC after IV administration and 4.5% after oral administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that fenbendazole was rapidly eliminated from plasma of pigs. The drug was rapidly absorbed after oral administration, but systemic bioavailability was low. PMID- 10803657 TI - Effects of endurance training on standard and signal-averaged electrocardiograms of sled dogs. AB - OBJECTIVE: To determine the effect of endurance training on QRS duration, QRS wave amplitude, and QT interval. ANIMALS: 100 sled dogs in Alaska. PROCEDURE: Dogs were examined in early September (before training) and late March (after training). During the interim, dogs trained by pulling a sled with a musher (mean, 20 km/d). Standard and signal-averaged ECG were obtained before and after training. RESULTS: Endurance training significantly increased mean QRS duration by 4.4 milliseconds for standard ECG (mean +/- SEM; 62.3 +/- 0.7 to 66.7 +/- 0.6 milliseconds) and 4.3 milliseconds for signal-averaged ECG (51.5 +/- 0.7 to 55.8 +/- 0.6 milliseconds) without changing body weight. Increase in QRS duration corresponded to a calculated increase in heart weight (standard ECG, 23%; signal averaged ECG, 27%). Signal-averaged QRS duration was correlated with echocardiographically determined left ventricular diastolic diameter for the X orthogonal lead (r = +0.41), Y orthogonal lead (r = +0.33), and vector (r = +0.35). Training also increased QT interval (234 +/- 2 to 249 +/- 2 milliseconds) and R-wave amplitude in leads II and rV2, increased peak-to-peak voltage and S wave amplitude in the Y orthogonal lead, and decreased Q-wave amplitude in the Y orthogonal lead. CONCLUSIONS AND CLINICAL RELEVANCE: Electrocardiographic changes reflected physiologic cardiac hypertrophy in these canine athletes in response to repetitive endurance exercise. The QRS duration increases in response to endurance exercise training and, therefore, may be of use in predicting performance in endurance activities. PMID- 10803656 TI - Quantitative urinalysis in healthy Beagle puppies from 9 to 27 weeks of age. AB - To evaluate indices of renal function in healthy, growing Beagle puppies from 9 to 27 weeks of age and to determine whether indices change with age during this period. Animals-6 healthy Beagle puppies. PROCEDURE: Urine collections were performed at 2-week intervals in puppies 9 to 27 weeks old. Daily excretion of urinary creatinine, protein, sodium, potassium, chloride, phosphorus, and calcium were determined, as were quantitative urinalyses including endogenous creatinine clearance, urine protein-to-creatinine ratios (UPr/C), and fractional clearances of sodium (FNa), potassium (FK), chloride (FCI), calcium (FCa), and phosphorus (FP). RESULTS: Significant differences among age groups were detected for endogenous creatinine clearance, and daily urinary protein, potassium, calcium, and phosphorus excretion. Significant differences also existed among age groups for UPr/C, FNa, FK, FCI and FP. Age-related effects fit a linear regression model for FNa, UPr/C, daily phosphorus excretion, and daily protein excretion. Quadratic regression models were judged most appropriate for endogenous creatinine clearance, FK, daily chloride excretion, and daily potassium excretion. Endogenous creatinine clearance measurements higher than adult reference ranges were observed from 9 to 21 weeks of age. The FNa, FK, FCI, FCa, and FP were slightly higher than those reported for adult dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Selected results of quantitative urinalyses in healthy 9- to 27-week-old Beagle puppies differ with age and differ from those measured in adult dogs. Diagnostic measurements performed in puppies of this age range should be compared with age-matched results when possible. PMID- 10803658 TI - Kinetic behavior of three preparations of alpha-tocopherol after oral administration to postpubertal heifers. AB - OBJECTIVE: To assess the kinetic behavior of 3 preparations of alpha-tocopherol (vitamin E) after oral administration to heifers. ANIMALS: 8 postpubertal Friesian heifers. PROCEDURE: A single oral bolus of 5,000 U of alpha-tocopherol in oil or encapsulated in liposomes or cyclodextrin was administered to each cow, using a 4 X 4 design with 8 days between treatments. Blood samples for kinetic analyses were obtained at various times for 168 hours after treatment. RESULTS: Mean (+/- SEM) maximal plasma concentrations of alpha-tocopherol were 4.86 +/- 0.49 microg/ml, 5.03 +/- 0.39 microg/ml, and 5.08 +/- 0.56 microg/ml after administration of oil, liposomal, and cyclodextrin preparations, respectively. Plasma concentrations peaked 21 to 34 hours after administration. The disappearance rate constant (Kd) was less after administration of alpha tocopherol encapsulated in liposomes, compared with the other 2 preparations. Area under the concentration versus time curve was greater after administration of either encapsulated form of alpha-tocopherol, compared with alpha-tocopherol in oil, but these differences were not significant. CONCLUSIONS AND CLINICAL RELEVANCE: The lower Kd determined for alpha-tocopherol encapsulated in liposomes suggests that this formulation may result in longer persistance of the vitamin in plasma than the other 2 preparations. Dietary supplementation with alpha tocopherol encapsulated in liposomes may enhance plasma availability of this vitamin in cattle and could be useful during periods of increased vitamin E requirements, such as parturition and early stages of life. PMID- 10803659 TI - The 9-bp deletion between the mitochondrial lysine tRNA and COII genes in tribal populations of India. AB - A 9-base-pair (bp) deletion located between the lysine tRNA (MTTK) and COII (MTCOX*2) genes in the human mitochondrial genome is a valuable marker for tracing population relationships. Previous research has shown that the 9-bp deletion is associated with two major clusters of control region sequences; one occurs in sub-Saharan Africa, while the other is associated with Asian populations and populations of Asian origin. We surveyed 898 individuals from 16 tribal populations in India and found 6 individuals with the 9-bp deletion. Sequences of the first hypervariable segment (HV1) of the mtDNA control region from these 9-bp deletion-bearing mtDNAs were compared to those previously reported from Asian and African populations. Phylogenetic analysis indicates three distinct clusters of tribal Indian 9-bp deletion mtDNA types. One cluster, found in northeast India, includes southeast Asian and Indonesian mtDNA types. The remaining two clusters appear to have unique origins in southern India. These data provide further evidence of past migrations from Asia into the northeast corner of the Indian subcontinent. PMID- 10803660 TI - RH blood groups and diabetic disorders: is there an effect on glycosylated hemoglobin level? AB - Recent cloning of RH genes has elucidated their structure, suggesting that RH proteins are part of an oligomeric complex with transport function in the erythrocyte. This observation prompted us to investigate a possible relationship between the RH system and the glycosylated hemoglobin level (Hb A(1c)) in diabetes. This compound is considered an important indicator- of glycemic control in diabetic disorders. We studied 278 subjects with non-insulin-dependent diabetes mellitus (NIDDM) from the population of Penne, Italy. Glycemic and glycosylated hemoglobin (Hb A(1c)) levels are associated with RH phenotype. Glucose and Hb A(1c) levels are increased in DCcEe subjects and decreased in ddccee subjects as compared to the mean values for other genotypes. Sex, age at onset of disease, duration of disease, and age of patients were also considered. Correlation analysis suggests that these variables influence glycemia directly and Hb A(1c) indirectly. The RH system, on the other hand, seems to influence the Hb A(1c) level directly. Preliminary data on 53 children with insulin-dependent diabetes mellitus (IDDM) from Sardinia seem to confirm the relationship between RH and Hb A(1c) observed in NIDDM. Since glycosylated hemoglobin is found inside red blood cells, the relationship between RH genetic variability and Hb A(1c) level suggests that RH proteins may influence glucose transport through red cell membrane and/or hemoglobin glycation. PMID- 10803661 TI - Distribution of the 3' VNTR polymorphism in the human dopamine transporter gene in world populations. AB - A polymorphism with a variable number of tandem repeats (VNTR) found in the 3' untranslated region of the human dopamine transporter gene (DAT1) was scored in unrelated individuals drawn from 10 geographically widely dispersed populations in order to assess this marker's usefulness in human population genetics. The populations that were analyzed in this study included 4 indigenous groups of Siberia, natives of North and South America, as well as Caucasian and Oceanic groups, most of which represented small-scale societies. A total of 5 DAT1 alleles were seen overall, but only in one Siberian population, the Altai-Kizhi, were all 5 present, and in the Native Americans of Colombia the locus was monomorphic. The most common allele, DAT1*10, ranged in frequency from 52% in Greeks to 100% in South Americans. The high frequency of the DAT1*10 allele (approximately 90%) among Mongoloid groups of north and east Asia distinguishes them from most Caucasian groups. The presence of the rare DAT1*7 allele in relatively high frequency (approximately 5%) among all Siberian groups suggests a close affinity with north Asian groups, especially Mongolians. The presence of the even rarer DAT1*13 allele in one Siberian population, the Altai-Kizhi, reflects this group's long historical contact with Mongolians. The results demonstrated that the DAT1 VNTR polymorphism is useful in investigating population relationships, and that rare alleles at this locus may be particularly valuable in understanding the extent of genetic affinity between neighboring groups and in situations where admixture is suspected. However, because of both the association and linkage of this VNTR locus with attention-deficit hyperactivity disorder (ADHD) in children, and its highly restricted polymorphism (usually 3 alleles) in most human groups, the possibility of selection constraints on the DAT1 gene cannot be ignored. PMID- 10803662 TI - GM, KM immunoglobulin allotypes and other serum genetic markers (HP, GC, PI, and TF) among South American populations living at different altitudes (Jujuy Province, Argentina): admixture estimates. AB - A total of 154 individuals belonging to three populations located at different altitude levels in northwest Argentina (San Salvador de Jujuy, 1,200 m; Tilcara, 2,500 m; Abra Pampa, 3,500 m) were studied for the GM, KM, HP, GC, PI and TF genetic systems. Individuals were selected on the basis of ethnocultural affiliation. Gene frequency values were found to be comparable to those reported for other South American populations. The populations studied showed a close genetic identity and an absence of interpopulation heterogeneity. Distribution of the GM phenotypes and haplotypes corresponds to historical data on human settlements in Jujuy Province. The presence of some alleles and the anthropological significance of the allele distribution are discussed, as are the effects of the admixture with Africans and Spaniards. The genetic pattern appears to be the result of a varying admixture due to the genetic isolation in populations located at various altitude levels. PMID- 10803663 TI - Plasma lipoprotein(a) distribution and its correlates among Samoans. AB - Plasma lipoprotein(a) [Lp(a)]-consisting of a disulfide-linked complex of apolipoprotein B and apolipoprotein (a)--levels are considered to be an independent risk factor for coronary heart disease. There are considerable ethnic group differences in the distribution of plasma Lp(a) levels that raise public health concerns. Although plasma Lp(a) distribution has been determined in various ethnic groups, no such information is available in Pacific Islanders. In this study we have determined the distribution and correlates of plasma Lp(a) in population-based samples of 361 American Samoans (145 men, 216 women) and 560 Western Samoans (265 men, 295 women), aged 20-70 years. Plasma Lp(a) levels were measured using a commercial enzyme-linked immunosorbent assay. The distribution of plasma Lp(a) levels in both groups was highly skewed with 73% and 65% of values in the 0-5 mg/dl range in American Samoans and Western Samoans, respectively. The mean (6.4 mg/dl) and median (2.2 mg/dl) Lp(a) levels in pooled Samoans were significantly lower when compared with other ethnic groups using the same measurement kit. Plasma Lp(a) correlated significantly with total and LDL cholesterol in both genders after correcting for the contribution of Lp(a) cholesterol, and with apolipoprotein B in women after the correction for Lp(a) apoB, but not with age, smoking, alcohol intake, or body mass index. Our data show that Samoans, Polynesians of Pacific Islands, have strikingly lower Lp(a) levels than all other reported population groups. These data are consistent with the hypothesis that genetic factors account for interethnic group variation in plasma Lp(a) levels. PMID- 10803664 TI - Analysis of HLA-DRB1 polymorphism in the Gidra of Papua New Guinea. AB - The genetic structure of the Gidra-speaking population inhabiting 13 villages in Papua New Guinea was investigated, based on the analysis of HLA-DRB1 polymorphism. Nei's fixation indices (F(IS), F(IT), and F(ST)) showed that the Gidra villages were genetically differentiated. The genetic distances significantly correlated with the geographic distances among the 13 villages. Thus, it is likely that a low intervillage migration rate has been maintained since the Gidra community was established. Correspondence analysis revealed that the Gidra, who belong to non-Austronesian-speaking groups, are genetically located at the intermediate point between the Aboriginal Australian groups and the Austronesian-speaking groups. Moreover, the HLA-DRB1*0802 allele, which has been observed in only two Polynesian groups (Austronesian-speaking groups) of Oceanian populations, was also found in the Gidra. These results suggest that the admixture of Austronesian and indigenous non-Austronesian groups beyond the linguistic boundary occurred partly in Papua New Guinea before Austronesian groups spread to the Pacific. PMID- 10803665 TI - Natural selection and reproductive behavior in Italy, 1930-1993. AB - We studied the relaxation of natural selection affecting the newborn population in Italy between 1930 and 1993 due to the decrease in the stillbirth (mortality) rate and the simultaneous changes in women's reproductive behavior (strategies). Results show that, apart from a drastic overall reduction, the stillbirth rate has varied among different groups of neonates. The present stillbirth rate of less than 5 per thousand, observed in 8 of the 20 phenotypic classes defined on the basis of maternal age at delivery and neonate birth order, most likely represents an unavoidable biological or genetic cost. A 9 per thousand stillbirth rate, about twice the potential minimum, exists among neonates born to women delivering at advanced age (> or = 35): the risk they face at the first or, in adverse living conditions, the fourth pregnancy could, however, be decreased by a policy aimed at countering the tendency to delay maternity and improving antenatal care in economically disadvantaged areas of the country. PMID- 10803666 TI - Population evolution in 20th-century Easter Island: endogamy and admixture. AB - We studied the 20th-century evolution of the Rapanui population of Easter Island, the most geographically isolated in the world, to analyze the current process of admixture. Using parochial birth records, we determined origin of the birth parents based on their surnames. The origin of parents reveals two stages of population evolution: endogamy, due to the isolation of the island, but with a strong rejection of isonymous marriages; and admixture, beginning in 1965 with the opening of the island to the rest of the world. We used Lasker's coefficient (Lasker's Ri) and the Shannon-Weaver coefficient of diversity (H) to characterize both stages. The gene flow evaluated from admixture has increased significantly since 1965. Births from exogamous unions represented 3.5% of total births from 1937 to 1965. increased to 43.2% between 1966 and 1980, and constituted 50.8% of all births between 1981 and 1996. PMID- 10803667 TI - Beta-globin gene haplotypes and alpha-thalassemia analysis in Babinga pygmies from Congo-Brazzaville. AB - We analyzed beta-globin gene cluster haplotypes and deletional alpha+-thalassemia (-alpha3.7kb) in 54 Babinga pygmy subjects from Congo-Brazzaville. The beta(S) globin gene frequency was 0.065 and that of the deletional alpha-globin gene ( alpha3.7kb) was 0.29. Eighty-five percent of the beta(S) chromosomes and 13% of the beta(A) chromosomes were associated with the Bantu haplotype, 10% of beta(A) chromosomes with the Senegal haplotype, and the remaining beta chromosomes with atypical haplotypes. None of the chromosomes were of the Benin haplotype. These results are clearly of anthropological and evolutionary interest. They also support earlier observations that alpha+-thalassemia is prevalent at a high frequency in African populations. PMID- 10803668 TI - A whole-body registration-free navigation system for image-guided surgery and interventional radiology. AB - RATIONALE AND OBJECTIVES: To develop and test an image-guided navigation system in which the base of reference is taken from the imaging modality, here, a helical CT scanner. METHODS: An optical digitizer together with a calibration device is used to measure the transformation matrix between the digitizer reference system and a CT reference system. During intervention, it tracks radiological and surgical tools with tool references. A specific software visually integrates the current tool position with the corresponding image information. In vitro accuracy tests were performed. RESULTS: With helical CT, freehand positioning accuracy was 1.9 +/- 1.1 mm (mean +/- SD) in vitro (n = 718). CONCLUSIONS: The navigation system developed by the authors appears to be feasible for radiological interventions as well as for minimally invasive surgery. It is not limited to a certain procedure, can be used in every region of the body, and is functional after imaging. Intraprocedural scans can be integrated immediately. PMID- 10803669 TI - Results of electrochemical therapy of colorectal liver metastases in rats followed up by MRI. AB - RATIONALE AND OBJECTIVES: Direct-current or electrochemical therapy is an alternative method for local tumor therapy. Until recently, it was mainly applied in China and was relatively unknown in the Western world. This study examines the feasibility and effectiveness of applying direct-current therapy in liver metastases of colorectal carcinomas in an animal model. METHODS: Liver metastases were implanted in 47 BDIX rats by subcapsular injection of cells from a colorectal strain (DHD/K12). The success rate of implantation and the size of the tumors were determined after 3 weeks by MR imaging (T2-weighted turbo spin-echo images; relaxation time 1,800 ms, echo time 80 ms). The direct-current therapy was applied by one platinum electrode placed in the center of the tumor and four at the periphery of the tumor. Suitable therapy parameters were established in a pilot study by comparing four different methods of direct-current therapy with a control group. The methods varied with respect to the electrode polarity and the applied voltage. In a second series of investigations, tumor growth was monitored by MR imaging 3 and 5 weeks after therapy. RESULTS: The tumor implantation rate was 92.6% at a tumor cell concentration of 8 x 10(5)/mL. The most effective therapy method was achieved by placing an anode at the center of the tumor and four cathodes at the periphery, with an applied charge of 80 C/cm3. Complete tumor necrosis was observed in 54% of cases. In the follow-up measurements, the mean tumor diameter was 0.65 cm at 3 weeks after therapy and 0.76 cm at 5 weeks after therapy. In comparison with the control group (1.08 and 1.53 cm, respectively), this represented a significant reduction in tumor growth rate. CONCLUSIONS: This study is the first to demonstrate an antitumoral effect of direct-current therapy on liver metastases of colorectal cancer in rats. PMID- 10803670 TI - A closed-chest pulmonary artery occlusion/reperfusion model in the pig: detection of experimental pulmonary embolism with MR angiography and perfusion MR imaging. AB - RATIONALE AND OBJECTIVES: To establish a pig model suitable for imitating pulmonary emboli to facilitate research in the diagnosis of pulmonary embolism. METHODS: Thirteen animals were anesthetized, mechanically ventilated, and subjected to pulmonary artery catheterization initiated from the right external jugular vein. With the use of a Swan-Ganz catheter, repetitive occlusion/reperfusion maneuvers were done at different locations of the pulmonary arterial tree. Conventional pulmonary angiography, MR angiography, and perfusion MR imaging were performed. RESULTS: The model remained hemodynamically stable throughout the 13 experiments, without any significant difference between the blood pressure measurements at the start and at the end of the right-heart and pulmonary artery catheterizations. In each of the nine animal experiments that investigated MR imaging, four of four using perfusion MR imaging (proximal and distal occlusions) and five of five using MR angiography (larger pulmonary artery occlusions), all repeated pulmonary artery occlusions were successfully performed (reproducibility of 100%). CONCLUSIONS: The closed-chest pulmonary artery occlusion/reperfusion model in the pig allowed repetitive, controlled imitations of pulmonary emboli at different levels of the pulmonary artery in the same experiment. MR angiography and perfusion MR imaging were adequate to detect the pulmonary artery occlusions and the nonperfused lung regions, respectively. The model may be a helpful tool for future research in this field. PMID- 10803671 TI - Comparison of double-contrast CT arthrography image quality with nonionic contrast agents: isotonic dimeric iodixanol 270 mg I/mL and monomeric iohexol 300 mg I/mL. AB - RATIONALE AND OBJECTIVES: The authors evaluated the image quality on delayed CT arthrography images with the use of the nonionic dimeric contrast agent, iodixanol 270 mg I/mL, compared with iohexol 300 mg I/mL. METHODS: A total of 132 patients with shoulder pain were included in a randomized, parallel, double-blind study. Sixty seven patients received iodixanol and 65 patients received iohexol. Patients underwent two CT-arthrography examinations: the first was performed 20 to 30 minutes after contrast injection and the second, 50 to 70 minutes after contrast injection. Data from 31 patients were excluded from the efficacy analysis. The overall quality of CT arthrography images was graded into four categories: excellent, good, moderate, and bad. RESULTS: The overall quality of delayed CT arthrography images was significantly better in the iodixanol group (P = 0.001, alpha = 0.05). On the first CT examination, image quality was good or excellent in 88% of the cases in the iodixanol group and in 96.1% in the iohexol group. The results on the delayed CT arthrography examination indicated that image quality was good or excellent in 88% of the cases when iodixanol was used and in 63.5% when iohexol was used. CONCLUSIONS: The overall quality of delayed images was significantly better with iodixanol than with iohexol. PMID- 10803672 TI - Fat- and water-selective MR cine imaging of the human heart: assessment of right ventricular dysplasia. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to develop and implement MR sequences for chemical shift-selective breath-hold cine imaging of the heart. Fibroadipose conversion of myocardium in cases suspected of right ventricular dysplasia should be revealed in fat- and water-selective MR images of high quality. METHODS: Frequency-selective saturation of one chemical shift component was applied in modified k-space-segmented, electrocardiography-gated sequences, allowing high-quality cine imaging of the human heart in a single breath-hold. Phantom studies and human examinations in eight normal subjects (aged 24-62 years) and in seven patients (aged 31-47 years) with suspected right ventricular dysplasia were performed. The patients showed suspicious findings, such as a dyskinetic and dilated right ventricle combined with ventricular arrhythmia, and underwent MR imaging after exclusion of other possible reasons (eg, coronary artery disease or pulmonary hypertension). RESULTS: High selectivity to the desired chemical shift component was confirmed by test measurements in a phantom containing water and lipids. In the human subjects, minor problems with magnetic field inhomogeneities appeared in the thoracic walls only. Four patients with suspected right ventricular dysplasia showed clearly abnormal signal behavior of the right myocardial wall in both fat- and water-selective cine images. Bright transmural structures were exhibited in fat-selective images, but the origin of the fat (epicardium or infiltrated myocardium) was often difficult to assess. CONCLUSIONS: Right ventricular areas with fibrosis and fatty degeneration often show normal signal intensity in standard T1-weighted images but can be differentiated from normal tissue by the new chemical shift-selective breath-hold cine techniques. PMID- 10803673 TI - Quantitative estimation of attenuation in ultrasound video images: correlation with histology in diffuse liver disease. AB - RATIONALE AND OBJECTIVES: To determine the relationship between the attenuation of backscatter intensity in B-scan images of the liver and diffuse liver disease in order to assess the usefulness of this method in providing quantitative objective characterization of diffuse liver diseases in general and in fatty liver in particular. METHODS: Twenty-four healthy volunteers and 28 patients with elevated liver enzyme levels who underwent liver biopsy were included in this study. An automatic far-field slope (FFS) algorithm that estimates the decrease in amplitude of the backscattered echo as a function of beam depth was implemented on the noncompensated image that was acquired on a commercial phased array ultrasound system fitted to a custom-built interface card. The images were processed at a work-station. All scans were acquired repeatedly, read, and graded blindly by experienced ultrasound radiologists. Histology obtained via needle biopsy was reviewed without knowledge of the ultrasound findings. RESULTS: Analysis of the FFS data for fatty infiltration in all patient groups yielded a sensitivity of 67%, a specificity of 77%, a positive predictive value (PPV) of 77%, negative predictive value (NPV) of 67%, and an accuracy of 71%. The mean score of the ultrasound reviewers showed a sensitivity of 82%, a specificity of 66%, a PPV% of 68%, an NPV of 81%, and an accuracy of 72%. Normal FFS values (false-negative) were found in five patients with proved fatty infiltration. All of these patients had coexistent moderate to severe hepatic inflammation. However, FFS data in patients with uncomplicated (pure) fatty infiltration revealed a sensitivity of 100%, a specificity of 80%, a PPV of 89%, an NPV of 100%, and an accuracy of 92%. The best ultrasound score yielded a sensitivity of 100%, a specificity of 60%, a PPV of 80%, an NPV of 100%, and an accuracy of 85% in the same patients. CONCLUSIONS: The data demonstrate an excellent sensitivity (100%) of the FFS values in patients with uncomplicated fatty infiltration. This was also the only group of patients in whom the FFS score was superior to the radiologists' best score. The FFS method can be used as a tool to follow up the response to a clinical or research treatment and to obtain standardization of pattern interpretation independently of the individual reader. PMID- 10803674 TI - A physical model for dual-energy X-ray absorptiometry--derived bone mineral density. AB - RATIONALE AND OBJECTIVES: Dual-energy X-ray absorptiometry (DXA)-derived areal bone mineral density (BMD) is an established predictor of osteoporotic fractures and reflects bone strength as well. The goal of this study was to develop and validate a physical model for appropriate interpretation of BMD. METHODS: DXA and peripheral quantitative computed tomography investigations of the distal tibia (n = 45), proximal tibia (n = 12), distal femur (n = 26), and distal radius (n = 34) were carried out. The DXA-derived BMD was analytically modeled as a nonlinear function of volumetric bone mineral apparent density and the cross-sectional area (eCSA) of given bone; ie, BMD(mod) = apparent BMD x square root of eCSA. RESULTS: At every measured skeletal site, the relationship between BMD and BMD(mod) was systematically stronger than that observed separately between BMD and apparent BMD or cross-sectional area. The models (r2) explained 85%, 94%, 87%, and 74% of the variability in BMD at the distal tibia, proximal tibia, distal femur, and distal radius, respectively. CONCLUSIONS: The mutual contributions of bone density and size to BMD can vary to some extent in a site-dependent fashion. This dual nature of BMD on one hand provides a reasonable mechanical explanation for why BMD is a good surrogate of bone strength and a predictor of osteoporotic fractures but on the other hand, complicates its detailed interpretation. PMID- 10803675 TI - Virtual CT cystoscopy: color mapping of bladder wall thickness. AB - RATIONALE AND OBJECTIVES: To improve the conspicuity of bladder tumors in a virtual environment, we developed an algorithm for color mapping the thickness of the bladder wall. The purpose of this study was to demonstrate the feasibility of this algorithm as a component of virtual CT cystoscopy. METHODS: Five subjects with a history of superficial transitional-cell carcinoma of the bladder underwent helical CT scanning after insufflation of the bladder with air. Source images were transformed into three-dimensional models, and the thickness of the bladder wall was demarcated by using a new computer algorithm and a fixed color scale. Results were compared with those obtained by conventional cystoscopy. RESULTS: Three tumors, one site of benign wall thickening, and normal wall thickness were correctly identified by using axial source images and virtual cystoscopy with color mapping. CONCLUSIONS: Color mapping of bladder wall thickness is feasible and demonstrates both normal and thickened urothelium. Its value in identification of small or sessile tumors will require further trials. PMID- 10803676 TI - Phenotyping of drug-metabolizing enzymes in adults: a review of in-vivo cytochrome P450 phenotyping probes. AB - Cytochrome P450 phenotyping provides valuable information about real-time activity of these important drug-metabolizing enzymes through the use of specific probe drugs. Despite more than 20 years of research, few conclusions regarding optimal phenotyping methods have been reached. Caffeine offers many advantages for CYP1A2 phenotyping, but the widely used caffeine urinary metabolic ratios may not be the optimal method of measuring CYP1A2 activity. Several probes of CYP2C9 activity have been suggested, but little information exists regarding their use, largely due to the narrow therapeutic index of most CYP2C9 probes. Mephenytoin has long been considered the standard CYP2C19 phenotyping probe, but problems such as sample stability and adverse effects have prompted the investigation of potential alternatives, such as omeprazole. Several well-validated CYP2D6 probes are available, including dextromethorphan, debrisoquin and sparteine, but, in most cases, dextromethorphan may be preferred due to its wide safety margin and availability. Chlorzoxazone remains the only CYP2E1 probe that has received much study. However, questions concerning phenotyping method and involvement of other enzymes have impaired its acceptance as a suitable CYP2E1 phenotyping probe. CYP3A phenotyping has been the subject of numerous investigations, reviews and commentaries. Nevertheless, much controversy regarding the selection of an ideal CYP3A probe remains. Of all the proposed methods, midazolam plasma clearance and the erythromycin breath test have been the most rigorously studied and appear to be the most reliable of the available methods. Despite the limitations of many currently available probes, with continued research, phenotyping will become an even more valuable research and clinical resource. PMID- 10803677 TI - Known variant DPYD alleles do not explain DPD deficiency in cancer patients. AB - Dihydropyrimidine dehydrogenase (DPD) degrades over 80% of administered 5 fluorouracil (5FU), thereby regulating the efficacy of this commonly used anticancer agent. DPD activity is highly variable (8-21-fold) and individuals with reduced activity have a high risk of 5FU toxicity. DPYD encodes DPD protein and 13 different mutations have been reported in DPD-deficient subjects. However, the contribution of these variant genotypes to polymorphic DPD activity in vivo is not clear. The previously described DPYD mutations are contained in 10 exons. These 10 exons were sequenced in a cohort of cancer patients with reduced (n = 23) or normal (n = 14) DPD activity to determine the contribution of each variant allele to low DPD activity in vivo. Eight of the 13 previously defined DPYD mutations (G62A, delta TCAT295-298, C703T, G1003T, G1156T, delta C1897, G2657A, and G2983T) were not detected. A previously defined exon 13 mutation (G1601A) was detected in three individuals with reduced DPD activity. An exon 14 splice donor site mutation (intron14 G1A) was detected in a normal DPD activity individual. It was demonstrated that T85C, A1627G and G2194A are common polymorphisms. A novel exonic mutation (T1679G) was detected in a patient with reduced DPD activity and 5FU toxicity. In addition, three novel common polymorphisms were detected in introns 10 and 13. Only three patients did not have any mutations and 30 had multiple DPYD mutations in the regions examined. However, only 17% (4/23) of the patients with a low DPD phenotype have a molecular basis for reduced activity. Although novel DPYD variants have been identified in this study, the 17 DPYD mutations now described do not entirely explain polymorphic DPD activity and toxic response to 5FU. These data emphasize the complex nature of the molecular mechanisms controlling polymorphic DPD activity in vivo. PMID- 10803678 TI - Relationship between NAT1 genotype and phenotype in a Japanese population. AB - NAT1, which biotransforms many carcinogens, is genetically polymorphic. This polymorphism has been postulated as a mechanism for susceptibility differences in cancer, possibly due to NAT1 activity differences. However, the relationship between NAT1 genotype and phenotype is not clear. In our study of 110 Japanese, the frequency of the NAT1*10 allele (0.53, 95% confidence interval 0.46-0.59) was higher than others have observed in Caucasians (0.16). From genotype frequency studies, 26.4% of the subjects belonged to the NAT1*10/*10 genotype, 53.6% to the NAT1*4/*10 genotype and 20% to the NAT1*4/*4 genotype. Neither NAT1*3 nor NAT1*11 genotype was seen in these subjects. In female subjects, we found higher NAT1 activity in NAT1*4/*10 subjects than in NAT1*4/*4 subjects (n = 49; 2.63 versus 2.16 nmol/min/mg protein). NAT1 activity-difference between NAT1*4/*10 and NAT1*10/*10 was not statistically significant. Thus, not only the presence of NAT1*10 allele, but also other factors are suspected of increasing NAT1 activities. After full sequencing of 10 subjects, five individuals having the highest activities and five individuals having the lowest activities, we found NAT1*18A and NAT1*18B to be in the high activity group and the low activity group, respectively. The genotypes containing these variants were heterozygous, i.e. NAT1*4/*18A and NAT1*4/*18B. Due to rare frequencies of these variants, they cannot be considered as other effective, genetic factors on NAT1 activity. Age and tobacco smoking did not affect the relationship between NAT1 genotype and phenotype. PMID- 10803679 TI - NAT2 polymorphism associated with plasma glucose concentration in Canadian Oji Cree. AB - The prevalence of type 2 diabetes in the Oji-Cree of Northern Ontario is among the highest of any population in the world. We previously demonstrated that markers D8S264 and D22S683 were both linked and associated with type 2 diabetes in the Oji-Cree. Among the possible candidate genes for type 2 diabetes and related traits on chromosomes 8p and 22q were NAT2 and CYP2D6, respectively. We thus explored the possible association of NAT2 and CYP2D6 alleles and diabetes related traits in a sample of 112 Oji-Cree subjects with type 2 diabetes and 481 Oji-Cree subjects without type 2 diabetes. We found no difference in the allele and genotype frequencies of the NAT2 G191A, C282T, C481T, G590A, A803G and G857A, and the CYP2D6 G1934A polymorphisms between Oji-Cree subjects with and without type 2 diabetes. However, we found a significant association between the NAT2 C282T polymorphism and plasma fasting glucose concentration. Specifically, NAT2 282T/T homozygotes had significantly higher plasma fasting glucose than 282C/C homozygotes, and heterozygotes had intermediate levels of this trait. Thus, variation in NAT2 or CYP2D6 was not associated with the presence of type 2 diabetes, and would not be causative for this phenotype in Oji-Cree. However, NAT2 might be a 'modifier gene' affecting the level of glycaemia in non-diabetic subjects. PMID- 10803680 TI - Role of polymorphic CYP2E1 and CYP2D6 genes in NNK-induced chromosome aberrations in cultured human lymphocytes. AB - Polymorphisms in genes of xenobiotic-metabolizing enzymes are largely responsible for interindividual differences in ability to activate and detoxify genotoxic agents and therefore may influence individual susceptibility to environmental cancer. The tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone (NNK), requires metabolic activation by cytochrome P450 (CYP) enzymes to generate DNA-reactive intermediates that induce mutations and cancer. In the current study, we investigated the role of the polymorphic CYP2E1 and CYP2D6 genes in the genotoxicity of NNK using the tandem-probe fluorescence in-situ hybridization (FISH) chromosome aberration assay as a marker. Our results, using whole blood cultures from 39 volunteers, indicated that NNK (0.12, 0.24 or 0.72 mM) induced a concentration-dependent increase in the frequency of chromosome aberration. The potential role of CYP2E1 and CYP2D6 in NNK-induced genetic damage in cultured human lymphocytes was characterized using specific CYP inhibitors. Treatment of blood cultures with 25 microM diethyldithiocarbamate (DDC), a specific CYP2E1 inhibitor, or 0.5 microM quinidine, a specific CYP2D6 inhibitor, simultaneously with NNK, significantly decreased NNK-induced chromosome aberration. We also studied the role of CYP2E1 and CYP2D6 allelic variants on NNK induced chromosome aberration. Our results indicate that NNK induced a significantly higher level of chromosome aberration in cells with the CYP2E1 WT/*5B genotype compared to cells with the CYP2E1 WT/WT. In contrast, no difference in NNK-induced chromosome aberration was observed between cells with the CYP2D6 extensive metabolizers compared to cells with the CYP2D6 poor metabolizer genotypes. These results underscore the important role of polymorphic metabolizing genes in influencing the genotoxic responses to environmental mutagens and provide support to the reported findings linking CYP2E1 polymorphism to smoking-related lung cancer. PMID- 10803681 TI - Genomic organization of the UGT2b gene cluster on human chromosome 4q13. AB - The UDP-glucuronosyltransferases (UGTs) comprise a large family of proteins capable of detoxifying a wide variety of both endogenous and exogenous substrates. The primary function of this gene superfamily is to catalyze the glycosylation of substrates such as biogenic amines, steroids, bile acids, phenolic compounds and various other pharmacologically relevant compounds, including numerous carcinogens, toxic environmental pollutants and prescription drugs. This conjugation increases the solubility of these compounds, allowing them to be excreted more readily through hepatic or renal mechanisms. This paper describes the genomic characterization and chromosomal localization of three UGT2B genes which together comprise part of a large cluster of related sequences, including pseudogenes found on human chromosome 4q13. A genomic map spanning approximately 500-1000 kb of this region reveals the presence of three previously described UGT2B genes, at least two previously uncharacterized pseudogenes and a significant number of remnant gene fragments and places UGT2B4 between UGT2B7 and UGT2B15. Additionally, access to a large reference DNA bank allowed us to calculate allele frequencies for two UGT2B SNPs: D85R in UGT2B15 and Q458D in UGT2B4 amongst 803 unrelated individuals representing five ethnic populations. The data presented here suggest a recent evolutionary history of gene duplication, mutation and rearrangement. Furthermore, they suggest that a re evaluation of the current description of the UGT2B gene family with respect to the number of specific genes, degree of allelic diversity and molecular evolution may be necessary. PMID- 10803682 TI - Histamine N-methyltransferase pharmacogenetics: association of a common functional polymorphism with asthma. AB - Histamine is involved in the pathophysiology of asthma, and histamine N methyltransferase (HNMT) plays the dominant role in histamine metabolism in human bronchial epithelium. Levels of HNMT activity in human tissues are controlled, in part, by inheritance. A common C314T polymorphism within the HNMT gene results in a Thr105Ile change in encoded amino acid, and the T314 allele is associated with decreased levels of both HNMT enzymatic activity and immunoreactive protein. Therefore, presence of the T314 allele would be expected to result in reduced histamine metabolism and increased bronchoconstriction. We characterized this common, functionally significant polymorphism in DNA samples from 237 randomly selected Caucasian control subjects and 192 samples from Caucasian asthmatic patients. Allele frequencies for the T314 HNMT allele were 0.08 in the control samples and 0.14 in samples from Caucasian asthmatic patients (odds ratio = 1.9, P < 0.01), indicating a significant increase in the frequency of subjects with low HNMT activity among asthmatics. The association between a common, functionally significant genetic polymorphism for HNMT and asthma suggests that individual variation in histamine metabolism might contribute to the pathophysiology and/or response to therapy of this disease. PMID- 10803683 TI - Torsemide metabolism by CYP2C9 variants and other human CYP2C subfamily enzymes. PMID- 10803684 TI - Rapid analysis of GSTM1, GSTT1 and GSTP1 polymorphisms using real-time polymerase chain reaction. PMID- 10803685 TI - Can caffeine metabolism be used as an in-vivo probe for human flavin-containing monooxygenase activity? PMID- 10803686 TI - Vesicular transport as a new paradigm in short-term regulation of transepithelial transport. AB - The vectorial transepithelial transport of water and electrolytes in the renal epithelium is achieved by the polarized distribution of various transport proteins in the apical and basolateral membrane. The short-term regulation of transepithelial transport has been traditionally thought to be mediated by kinetic alterations of transporter without changing the number of transporters. However, a growing body of recent evidence supports the possibility that the stimulus-dependent recycling of transporter-carrying vesicles can alter the abundance of transporters in the plasma membrane in parallel changes in transepithelial transport functions. The abundance of transporters in the plasma membrane is determined by net balance between stimulus-dependent exocytic insertion of transporters into and endocytic retrieval of them from the plasma membrane. The vesicular recycling occurs along the tracts of the actin microfilaments and microtubules with associated motors. This review is to highlight the importance of vesicular transport in the short-term regulatory process of transepithelial transport in the renal epithelium. In the short-term regulation of many other renal transporters, vesicular transport is likely to be also involved. Thus, vesicular transport is now emerged as a wide-spread general regulatory mechanism involved in short-term regulation of renal functions. PMID- 10803687 TI - Little role of anti-gB antibodies in neutralizing activity of patient's sera with human cytomegalovirus (HCMV) infection. AB - Human cytomegalovirus (HCMV) gB is known to play important roles in cell surface attachment, virion penetration, spread of infection from cell to cell, and provocation of neutralizing antibody. This study was performed to determine the role of anti-HCMV gB antibody in overall neutralizing response in patients with HCMV infection and healthy control with past infection. HCMV gB was stably expressed in 293 cells. With the stable cell line expressing gB as a specific immunosorbent, anti-gB antibody was removed from the current and past HCMV infected sera and the remaining neutralizing activity was measured by plaque assay. It was shown that 19-50% of the total virus-neutralizing activity of sera with past HCMV infections was derived from anti-gB antibody, but anti-gB antibody had little effect on the total serum virus-neutralizing activity in patients currently infected with HCMV. This result suggests that neutralizing antibody to HCMV gB may reflect disease status. PMID- 10803688 TI - Assessment of coronary flow reserve with transthoracic Doppler echocardiography: comparison with intracoronary Doppler method. AB - To evaluate the feasibility and usefulness of transthoracic Doppler echocardiography (TTDE) as a non-invasive method in recording distal anterior descending (LAD) coronary flow velocity, we compared coronary flow reserve (CFR) measured by TTDE with measurements by intracoronary Doppler wire (ICDW). Twenty one patients without LAD stenosis were studied. ICDW performed at baseline and after intracoronary injection of 18 microg adenosine. TTDE was performed at baseline and after intravenous adenosine (140 microg/kgmin for 2 min). Adequate Doppler recordings of coronary flow velocities during systole were obtained in 14 of 21 study patients (67%) and during diastole in 17 (81%) patients. Baseline and hyperemic peak diastolic flow velocities measured by TTDE were significantly smaller than those obtained by ICDW (p<0.05). However, diminishing trends of diastolic and systolic velocity ratio after hyperemia were similarly observed in both methods. CFR obtained by TTDE (3.0+/-0.5), was higher than the value calculated by ICDW (2.5+/-0.4). There were significant correlations between the values obtained by the two methods (r=0.72, p<0.01). It is concluded that TTDE is a feasible method in measuring coronary flow velocity and appears to be a promising non-invasive method in evaluating CFR. PMID- 10803689 TI - Plasminogen activator inhibitor-1 4G/5G promoter polymorphism and coagulation factor VII Arg353-->Gln polymorphism in Korean patients with coronary artery disease. AB - An increased risk for arterial thrombosis is associated with high plasma levels of coagulation and fibrinolytic factors such as PAI-1 and FVII. In this study, the 4G/5G polymorphism in the promoter of PAI-1 gene and Arg353-->Gln polymorphism in the FVII gene were analysed in 139 normal adults and 158 patients with coronary artery disease (CAD), and their association with plasma lipid traits was investigated. There were no significant differences in the allele frequencies of PAI-1 and FVII polymorphisms between control and patient groups. The allelic distributions of both polymorphisms in Koreans were similar to those in Japanese but significantly different from those in Caucasians. In the CAD group, the 4G homozygotes of PAI-1 polymorphism showed significantly higher levels of total (p=0.0250) and LDL cholesterol (p=0.0335) with individuals having other genotypes. However, FVII polymorphism showed no association with lipid levels. In conclusion, the 4G/5G PAI-1 promoter polymorphism and Arg353-->Gln FVII polymorphism are not major genetic risk factors for CAD in Koreans. However, 4G allele of PAI-1 polymorphism revealed to be associated with the levels of cholesterol, especially LDL cholesterol levels in CAD patients. PMID- 10803690 TI - Comparison of respiratory indices in predicting response to high frequency oscillatory ventilation in very low birth weight infants with respiratory distress syndrome. AB - To evaluate the predictive values of oxygenation index (OI), arterial-alveolar oxygen tension ratio (a/APO)2, and alveolar-arterial oxygen gradient ((A-a)DO2) for early recognition of responsiveness to high frequency oscillatory ventilation (HFOV) in very low birth weight infants with respiratory distress syndrome (RDS), 23 infants who received HFOV treatment for severe RDS after failing to be improved with conventional mechanical ventilation from July 1995 to February 1998 were included. Twelve infants survived with HFOV (Responder group), while 11 infants could not maintain oxygenation with HFOV and died (Non-responder group). Clinical record (of each patient) were retrospectively reviewed and compared with the respiratory indices. Mean (A-a)DO2 was significantly lower in the responder group than in the non-responder group at 2 hr after HFOV (p=0.024), and the difference was more remarkable at 6 hr (p=0.005). Death in the patient with (A a)DO2 over 350 at 2 hr after HFOV therapy was 100% in sensitivity and 80% in specificity. The earliest significant difference of mean a/APO2 between two groups was noted at 6 hr after HFOV treatment (p=0.019). OI showed no significant differences between two groups. In summary, (A-a)DO2 was the most effective and sensitive respiratory index for predicting the responsiveness to HFOV in infants with severe RDS providing due as early as 2 hr. PMID- 10803691 TI - DNA ploidy patterns in gastric adenocarcinoma. AB - To assess the value of DNA ploidy, flow cytometric analysis was performed on unfixed fresh materials obtained from 86 patients with gastric cancer who underwent stomach resection. We evaluated the DNA content of gastric carcinoma cells from four different sites and compared it with Ki-67 proliferating activity, and other pathologic parameters. The incidence of aneuploid and diploid was similar (48.8% vs. 51.1%). Early gastric carcinoma showed a higher rate of the diploid pattern (75%) compared to that of advanced gastric carcinoma (47.3%). DNA diploidy was noted increasingly in diffuse-type tumors according to Lauren, in signet ring cell type tumor according to WHO classification and in poorly differentiated tumors (p<0.05). Well and moderately differentiated carcinomas revealed the aneuploid pattern more frequently than poorly differentiated tumors. The aneuploidy was associated with high S phase fraction and high proliferative index. Aneuploidy was noted in the mucosa adjacent to the tumor (26%), in the close normal-looking mucosa (7%) and in the remote normal-looking mucosa (3%). This result suggest the possible role of field cancerization in the development of gastric adenocarcinoma. PMID- 10803692 TI - Change of telomerase activity in rectal cancer with chemoradiation therapy. AB - Telomerase, an enzyme associated with cellular immortality, is expressed by most malignant cells and is inactive in most normal somatic cells, with the exception of proliferative stem cells, germ cells and activated lymphocytes. Measuring telomerase activity clinically may provide useful diagnostic and prognostic information of cancer. The purpose of this study was to investigate the change in telomerase activity following chemoradiation in rectal cancer, which almost always produces positive enzymatic activity. A total of 24 tumor tissue samples were used in this study, consisting of 12 paired specimens before and 4 weeks after chemoradiation. Telomerase activity was determined by PCR-based telomeric repeat amplification protocol (TRAP) assay. The telomerase activity was positive in 10 out of 12 patients (83%) in pre-irradiated and post-irradiated states. The levels of telomerase activity was decreased in 8 out of 10 patients after chemoradiation (80%) and two cases showed no change in enzymatic activity. One case showed no activity in either sample. The other case showed no enzymatic activity in the pre-irradiated sample, but showed weak activity in the post irradiated sample. These data indicate that telomerase activity in rectal cancer is reduced after neoadjuvant chemoradiation therapy, possibly suggesting a mechanism of downstaging following chemoradiation therapy in cancer. PMID- 10803693 TI - CT-guided celiac plexus block for intractable abdominal pain. AB - Treatment of intractable abdominal pain due to inoperable intraabdominal malignancy is important, and the ineffectiveness of pharmacological agents has led many investigators to recommend chemical neurolysis of the celiac ganglions as a treatment. The author describes the technique and results of celiac plexus neurolysis under CT-guidance with various approach routes, including anterior, posterior and transaortic routes. Twenty-eight patients, ranging in age from 36 to 82 years, have been treated with this procedure. All had inoperable or recurred intraabdominal malignancies and suffered from intractable upper abdominal pain and/or back pain. The author performed the procedure using absolute alcohol by an anterior approach (n=18), posterior approach (n=6) and transaortic approach (n=4). Pain was rated according to a visual analog scale before and after the procedure to gauge treatment success. No major complications occurred. Mild hypotension occurred in five patients (18%) and transient diarrhea in six patients (21%). Twenty-one (75%) of the 28 patients had some relief of pain and 17 of these patients (61%) had good relief of pain after the procedure. The results support that CT-guided celiac plexus block with alcohol is a safe and effective means of pain control in patients with intraabdominal malignancy. PMID- 10803694 TI - HFE gene mutation, C282Y causing hereditary hemochromatosis in Caucasian is extremely rare in Korean population. AB - Hereditary hemochromatosis (HFE), which affects 1 in 400 and has an estimated carrier frequency of 1 in 10 individuals in Western population, results in multiple organ damage caused by iron deposition, and is treatable if detected early. C282Y mutation in HFE gene has been known to be responsible for the most hereditary hemochromatosis cases and 5-10% of white subjects are heterozygous for this mutation. However, the prevalence of hemochromatosis in the Asian population was reported to be very low and ethnic heterogeneity has been suspected. The aim of our study was to determine the prevalence of heterozygosity and homozygosity for the C282Y HFE gene mutations in 502 unrelated Koreans. Results revealed that none of them had the mutant gene, suggesting a significant ethnic difference when compared with Caucasians. Our study excluded underlying possibility of hereditary hemochromatosis in Korean which could mimic the findings of alcoholic liver disease with iron overload or liver cirrhosis with chronic hepatitis C. PMID- 10803695 TI - Erythropoietin does not affect nitric oxide system in rats with chronic renal failure. AB - We investigated to see whether an altered role of nitric oxide (NO) system is involved in erythropoietin (EPO)-induced hypertension in chronic renal failure (CRF). Male Sprague-Dawley rats were five-sixths nephrectomized to induce CRF. Six weeks after the operation, EPO or vehicle was injected for another 6 weeks. Plasma and urine nitrite/nitrate (NOx) levels were determined. Expression of NO synthase (NOS) proteins in the aortae and kidneys were also determined. In addition, the isometric tension of isolated aorta in response to acetylcholine and nitroprusside was examined. Blood pressure progressively rose in CRF groups, the degree of which was augmented by EPO treatment. Plasma NOx levels did not differ among the groups, while urine NOx levels were lower in CRF groups. Endothelial NOS expression was lower in the kidney and aorta in CRF rats, which was not further affected by EPO-treatment. The inducible NOS expression in the kidney and aorta was not different among the groups. Acetylcholine and sodium nitroprusside caused dose-dependent relaxations of aortic rings, the degree of which was not altered by EPO-treatment. Taken together, EPO-treatment aggravates hypertension in CRF, but altered role of NO system may not be involved. PMID- 10803696 TI - Changes in biochemical bone markers during pregnancy and puerperium. AB - To elucidate the changes in bone turnover during pregnancy and puerperium, we measured serially the levels of serum osteocalcin and urine deoxypyridinoline (Dpy) as markers of bone formation and bone resorption, respectively, in 22 healthy women with normal pregnancy. Nineteen non-pregnant women served as control. The Dpy levels increased significantly at 16 weeks of pregnancy and remained elevated thereafter. The levels of osteocalcin, however, were significantly decreased at 16 weeks of pregnancy and elevated later at 6 weeks postpartum. Bone turnover ratio (Dpy/osteocalcin) continued to rise during pregnancy, but returned to control levels 6 weeks after delivery. Dpy levels and bone turnover ratio during puerperium tended to be higher in 17 breast-feeding women than those of 5 exclusive bottle-feeders. In conclusion, bone resorption begins to increase from the second trimester of pregnancy and calcium release from bone tissue might play a major role in calcium homeostasis during the whole period of pregnancy as well as during lactation. PMID- 10803697 TI - The detection of circulating breast cancer cells in peripheral blood by reverse transcriptase-polymerase chain reaction. AB - Some circulating cancer cells in the blood play a central role in the metastatic process and may have a major influence on patient progress. Their numbers can be very small and techniques for their detection need to be both sensitive and specific. Polymerase chain reaction (PCR) has been successfully used to detect small numbers of tumor cells in cancer. We used a reverse transcriptase polymerase chain reaction (RT-PCR) to detect circulating breast cancer cells in venous blood samples before operations and assessed cytokeratin-19 (CK-19) and cytokeratin-20 (CK-20) as target mRNA markers in the blood of healthy donors (n=6) and breast cancer patients (n=30) with American Joint Committee on Cancer stages 0 to IIIa. CK-19 mRNA was expressed in all blood samples of healthy donors and patients. But CK-20 was the only mRNA marker not detected in the blood from healthy donors. Seven of 30 (23%) venous blood isolates of breast cancer patients yielded a CK-20 mRNA with positive results. There was no correlating CK-20 mRNA expression with stage and axillary lymph node status. In conclusion, CK-19 showed no diagnostic value as a mRNA marker in the detection of circulating cancer cells by RT-PCR assay because this was expressed in the blood of healthy donors. CK-20 mRNA was an useful marker to detect circulating cancer cells in breast cancers. PMID- 10803698 TI - Clinical constellation of annular erythema associated with anti-Ro/La autoantibodies. AB - Annular erythema (AE) associated with anti-Ro (SSA) and/or La (SSB) autoantibody in patients with Sjogren syndrome (SS) or with SS/systemic lupus erythematosus overlap syndrome (SS/SLE), has recently been described in Orientals, and it may be a counterpart of annular skin lesion of the subacute cutaneous LE seen mostly in Caucasians. The author examined five Korean AE patients in respect to dinical diversity. In this small-sample study, subtle differences appeared between individual cases regarding the serologic features and the diagnoses of the disease. Among the five cases, four had circulating anti-Ro and anti-La antibodies, and one had only anti-La. Regarding the diagnosis, one was SS/SLE, two were primary SS, and the remaining two were only "AE associated with anti Ro/La antibody". There seem to be a wide clinical spectrum in the disease expression of AE associated with anti-Ro/La autoantibody than previously thought. PMID- 10803699 TI - Effects of decreased cerebral perfusion pressure on cerebral hemodynamics, brain cell membrane function and energy metabolism during the early phase of experimental Escherichia coli meningitis in the newborn piglet. AB - In this study, we tested the hypothesis that decreased cerebral perfusion pressure (CPP) induces cerebral ischemia and worsen brain damage in neonatal bacterial meningitis. Meningitis was induced by intracisternal injection of 10(9) colony forming units of Escherichia coli in 21 newborn piglets. Although CPP decreased significantly at 8 hr after bacterial inoculation, deduced hemoglobin (HbD), measured as an index of changes in cerebral blood flow by near infrared spectroscopy, did not decrease significantly. In correlation analyses, CPP showed significant positive correlation with brain ATP and inverse correlation with brain lactate levels. CPP also correlated positively with HbD and oxidized cytochrome aa3 (Cyt aa3) by near infrared spectroscopy. However, CPP did not show significant correlation with cerebral cortical cell membrane Na+,K+-ATPase activity, nor with levels of lipid peroxidation products. In summary, decreased CPP observed in this study failed to induce cerebral ischemia and further brain injury, indicating that cerebrovascular autoregulation is intact during the early phase of experimental neonatal bacterial meningitis. PMID- 10803700 TI - Clinicopathologic analysis of 124 biopsy-proven peripheral nerve diseases. AB - We reviewed dinical, histological and ultrastructural findings of 124 cases of sural nerve biopsy specimens to delineate the trends of peripheral nerve diseases in our institute. Eighty-one were men and 43 were women. We categorized them into five groups: specific diagnosis (66 cases, 53.2%), axonal degeneration type (47 cases, 37.9%), demyelinating type (4 cases, 3.2%), mixed axonal degeneration demyelinating type (6 cases, 4.8%) and normal (1 case, 0.9%). Cases with specific diagnosis included 21 inflammatory demyelinating polyneuropathy (15 chronic inflammatory demyelinating polyradiculoneuropathy, 6 Guillain-Barre disease), 13 hereditary motor and sensory neuropathy (7 Charcot-Marie-Tooth type I, 6 Charcot Marie-Tooth type II), 10 vasculitis, 6 toxic neuropathy, 4 leprosy, 3 diabetic neuropathy, 2 alcoholic neuropathy, 1 Fabry's disease and other specific diseases (5 cases). In our cases, the proportion of specific diagnoses was higher, while the proportion of demyelinating peripheral neuropathies and normal were lower than those of Western series. The results of this study indicate that 1) a dose clinicopathologic correlation is important to make a precise diagnosis of peripheral nerve biopsy, 2) Biopsy under strict indication may reduce unnecessary histologic examination, 3) There is no difference in disease pattern of peripheral neuropathy between Western people and Koreans. PMID- 10803701 TI - Defects in the differentiation and function of bone marrow-derived dendritic cells in non-obese diabetic mice. AB - Due to their high immunostimulatory ability as well as the critical role they play in the maintenance of self-tolerance, dendritic cells have been implicated in the pathogenesis of autoimmune diseases. The non-obese diabetic (NOD) mouse is an animal model of autoimmune type 1 diabetes, in which pancreatic beta cells are selectively destroyed mainly by T cell-mediated immune responses. To elucidate initiation mechanisms of beta cell-specific autoimmunity, we attempted to generate bone marrow-derived dendritic cells from NOD mice. However, our results showed low proliferative response of NOD bone marrow cells and some defects in the differentiation into the myeloid dendritic cells. NOD dendritic cells showed lower expressions of MHC class II, B7-1, B7-2 and CD40, compared with C57BL/6 dendritic cells. In mixed lymphocyte reactions, stimulatory activities of NOD dendritic cells were also weak. Treatment with LPS, INF-gamma and anti-CD40 stimulated NOD dendritic cells to produce IL-12p70. The amount of IL-12, however, appeared to be lower than that of C57BL/6. Results of the present study indicated that there may be some defects in the development of NOD dendritic cells in the bone marrow, which might have an impact on the breakdown of self tolerance. PMID- 10803702 TI - Pulmonary fibrosis in a steel mill worker. AB - We report a case of pulmonary fibrosis in a 32-year-old man, who had worked at a steel mill and who died of respiratory failure due to interstitial fibrosis despite vigorous treatment. He showed SLE-associated symptoms, such as pleural effusion, malar rashes, discoid rashes, arthritis, leukopenia, and positive antinuclear antibody and anti-histone antibody. However, he did not present anti DNA antibody. A thoracoscopic lung biopsy showed interstitial fibrosis, chronic inflammation and a small non-caseating granuloma in lung tissues, which could be induced by external agents such as metals. The manganese concentration in the lung tissue was 4.64 microg/g compared to 0.42-0.7 microg/g in the controls. The levels of other metals, such as iron, nickel, cobalt and zinc in patient's lung tissue were higher than those in the controls. The patient was probably exposed to Si and various metal dusts, and the lung fibrosis was related to these exposures. Exposure to Si and metal dusts should be sought in the history of any patient with SLE, especially in a male with pulmonary signs, and if present, exposure should be stopped. In the meantime, steps should be taken to ensure that workers exposure to Si and metal dusts in all environments have adequate protection. PMID- 10803703 TI - Extranasal T/NK-cell lymphoma presenting as intestinal diverticulum. AB - A case of intestinal angiocentric T/NK-cell lymphoma in a 58-year-old man is reported. The patient presented initially with panperitonitis because of perforation of sigmoid colon diverticulum. He underwent segmentectomy of involved bowel. Histologically, the intestinal wall showed diffuse infiltration of medium or large size lymphoma cells with angiocentric growth and necrosis. The lymphoma cells were CD56+, CD45RO+, CD3+, CD4-, CD8-, CD20-, and CD30- in paraffin sections with germline configuration of TCR-gamma gene, consistent with T/NK-cell lymphoma. Further staging revealed splenomegaly. Intestinal angiocentric T/NK cell lymphoma represents a distinct etiology of diverticulum with perforation. PMID- 10803704 TI - Adult T cell leukemia/lymphoma with lymphopenia in a Korean. AB - We experienced a case of adult T cell leukemia/lymphoma (ATLL) in a 48-year-old Korean female, who has never been abroad since birth and no history of blood transfusion. The patient had hypercalcemia and multiple lymphadenopathy. Histopathologic study of left cervical lymph node (LN) and bone marrow (BM) revealed that infiltrates of malignant lymphoid cells were composed of small, medium and large cells with pleomorphic nuclei. Smears of peripheral blood (PB) showed lymphopenia (16%) with the appearance of a few atypical lymphoid cells (less than 2%), but not the typical clover leaf cells seen in ATLL. Immunophenotypic study of LN and BM revealed T cell phenotype. PB showed increased CD4+ T cell (T(H), CD3/CD4+, 57%) and decreased CD8+ T cell counts (T(S), CD3/CD8+, 6.7%). The sera of the patient and her family were reactive for HTLV-I antibody. The specific sequences of pol, env, and tax of HTLV-I DNA were detected in the lymphoma cells and peripheral blood mononuclear cells (PBMC) using polymerase chain reaction. Ultrastructural examination of PBMC confirmed numerous type c virus particles in extracellular space. This case was an acute type of ATLL without overt leukemic features in PB. Despite chemotherapy and intensive conservative treatment, she died 3 months after admission. PMID- 10803705 TI - A case report of primary T-cell lymphoma of the liver. AB - The patient was a 50-year-old woman who presented intermittent mild fever with elevated liver enzymes for 12 years. The liver biopsy showed diffuse portal and sinusoidal involvement of lymphoid cells with minimal atypia and epithelioid histiocytic granuloma formation. Subsequent bone marrow biopsy showed lymphomatous involvement. The lymphocytes infiltrating the liver were reactive for T-cell marker and showed TCR gamma gene rearrangement. The patient was diagnosed as primary peripheral T-cell lymphoma of the liver. Indolent clinical course and resemblance with hepatitis were considered to be a rare and unique feature of this case. PMID- 10803706 TI - Myelomatous effusion with poor response to chemotherapy. AB - While pleural effusion in multiple myeloma is relatively infrequent, myelomatous pleural effusion is extremely rare. We experienced a 61-year-old woman with IgD lambda multiple myeloma and pleural effusion. The diagnosis was made originally by pleural biopsy, pleural fluid cytology and immunoelectropheresis of pleural fluid. Transient improvement of the pleural effusion was observed after administration of combination chemotherapy of vincristine, melphalan, cyclophosphamide, prednisone (VMCP)/vincristine, cyclophosphamide, adriamycin, prednisone (VCAP). Two months later, myelomatous pleural effusion recurred and no response to salvage therapy was observed. We reviewed the clinical feature of this case and literature concerning myelomatous pleural effusion. PMID- 10803707 TI - Weber-Christian disease presenting with proptosis: a case report. AB - Weber-Christian disease (WCD) is a rare inflammatory disease of adipose tissue, which is characterized by painful cutaneous nodules and constitutional symptoms. Although any area of the body containing fat can be affected by WCD, the involvement of retrobulbar fat is uncommon and proptosis is a rare presenting manifestation. We report a case who presented with proptosis of the right eye which is accompanied by painful subcutaneous nodules, high fever and myalgia. Biopsies of retrobulbar tissue and suprapubic nodule showed lobular panniculitis with mixed cellular infiltration, mainly composed of histiocytes and lymphocytes. He responded well to high-dose glucocorticoid. PMID- 10803708 TI - A case of myasthenia gravis proven by ultrastructural study. AB - Although light microscopic features of muscle are not pathognomonic in most cases of myasthenia gravis (MG), careful examination of neuromuscular junction by electron microscopy (EM) can reveal important clues for this disease. We report here a case of MG confirmed by EM study to emphasize that tissue diagnosis is still the best adjuvant to confirm the diagnosis. An 18-year-old female visited our hospital complaining of progressive muscle weakness for 3 years. She had difficulty in running, going upstairs and doing routine activities. Symptoms were aggravated with continuous work and resolved after rest. She had weakness of bilateral masseter and facial muscles and proximal portions of extremities without definite diurnal variation. Electromyography showed myopathic changes in proximal muscles of extremities. MG was considered but tensilon test was equivocal. Repetitive nerve stimulation tests revealed 20-30 percent decrease in responses to low and high rate stimulation. Muscle biopsy revealed selective type 2 atrophy. Ultrastructurally, abnormalities of neuromuscular junctions, i.e., wide primary synaptic cleft, and wide and shallow secondary synaptic clefts with mild myopathic features were present. These findings were pathognomonic for MG. Later, her symptoms were improved completely 3 months after thymectomy. The histologic finding of thymus was follicular hyperplasia. PMID- 10803709 TI - Overexpression of novel protein kinase C delta in BL6 murine melanoma cells inhibits the proliferative capacity in vitro but enhances the metastatic potential in vivo. AB - In this study we analysed the effect of overexpressing novel protein kinase C delta isoform (n-PKC delta) on melanin synthesis and metastatic potential in the highly metastatic BL6 murine melanoma cells. The proliferative capacity in vitro and into matrigel in vivo were also examined. Although murine melanocytes express the n-PKC delta isoform, BL6 cells do not express this isoform at levels detectable by Western blot analysis. In untransfected and transfected cells we also studied the effect of 12-O-tetradecanoylphorbol-13-acetate (TPA), a modulator of specific isoforms of PKC, and of bryostatin 1, a potent immunomodulator and antineoplastic drug and a partial agonist of PKC. Our results demonstrate a pivotal role for this isoform in melanin synthesis and the close relationship between n-PKC delta expression and its association with the particulate fraction, melanogenesis and metastatic potential. In fact, heterogeneous BL6 cells overexpressing n-PKC delta and all the clones isolated showed increased intracellular melanin and metastatic capacity. TPA and bryostatin 1 decreased n-PKC delta expression, the intracellular melanin level and metastatic capacity in both cell lines. Therefore both treatments were able to abolish the effects of overexpressing n-PKC delta. PMID- 10803710 TI - Effects of L-2-oxothiazolidine-4-carboxylate on the cytotoxic activity and toxicity of cyclophosphamide in mice bearing B16F10 melanoma liver metastases. AB - Glutathione (GSH) is the major non-protein thiol in cells that plays a critical role against damage from electrophilic agents such as alkylating drugs. Selective therapeutic GSH elevation in normal but not in tumour cells has been suggested as a means of protecting host tissues against more intense doses of chemotherapy. The present study investigated the response of B16 melanoma to treatment with the cysteine pro-drug L-2-oxothiazolidine-4-carboxylate (OTZ), alone and in combination with cyclophosphamide (CY). We found that OTZ decreased the GSH levels and proliferation rate of B16 melanoma cells in vitro, sensitizing them to the cytotoxic action of the activated metabolite of CY, acrolein (AC). In contrast to OTZ, the cysteine deliverer N-acetylcysteine (NAC) enhanced B16 melanoma cell proliferation by increasing GSH levels, and markedly decreased the sensitivity of these tumour cells to AC. In vivo studies showed the antitumoral activity of OTZ in B16 melanoma liver metastasis-induced mice, increasing their life span. We also observed that, whereas with CY treatment the GSH levels in peripheral blood mononuclear cells (PBMCs) were reduced and a dose-dependent leukopenia was produced, OTZ significantly increased PBMC GSH content, reducing toxicity and enhancing the survival of mice bearing established melanoma liver metastases treated with lethal dose CY. These results suggest a critical role for OTZ in protecting against alkylator agent-induced immunosuppression, which may allow the dose escalation of these cytostatic drugs to improve their therapeutic benefit in the treatment of malignant melanoma. PMID- 10803711 TI - The detection of tyrosinase mRNA in the peripheral blood of stage I melanoma patients is not of clinical relevance in predicting metastasis risk and survival. AB - The presence of tyrosinase mRNA in the peripheral blood cells of melanoma patients has been recently studied as a possible marker of haematogenous dissemination. However, considerable variations in the rates of detection have been noted. We determined the presence of tyrosinase mRNA-positive circulating cells using reverse transcriptase-polymerase chain reaction (RT-PCR) in 35 patients with stage I melanoma, two patients with stage II melanoma and two patients with stage III melanoma. Among the patients with stage 1, 13 were tested before and after surgery (< 1 h). Twenty healthy subjects served as negative controls. Out of the melanoma patients, the tyrosinase gene was expressed in three of the 52 samples tested. Tyrosinase mRNA was present in the circulating cells of only one patient with stage I melanoma after intra-congenital naevi resection. However, two other stage I patients developed rapidly lethal metastasis within the following 6 months, despite the lack of detectable tyrosinase mRNA. None of stage II patients were positive for the tyrosinase transcripts, while both patients with stage III melanoma showed enzyme expression. Our results confirm those of previous studies, showing that a small proportion of stage I melanoma patients have tyrosinase-positive circulating cells. Moreover, the lack of tyrosinase mRNA detection in the blood does not necessarily exclude metastatic progression. Therefore, this study indicates that the detection of tyrosinase mRNA-positive circulating cells by RT-PCR is not a predictive biomarker of a metastasis risk in patients with stage I melanoma. PMID- 10803712 TI - Analysis of melanoma cells in peripheral blood by reverse transcription polymerase chain reaction for tyrosinase and MART-1 after mononuclear cell collection with cell preparation tubes: a comparison with the whole blood guanidinium isothiocyanate RNA isolation method. AB - Melanoma cell detection in peripheral blood by tyrosinase reverse transcription polymerase chain reaction (RT-PCR) is usually performed on RNA isolated from whole blood using a guanidinium isothiocyanate (GITC)/phenol extraction method or from Ficoll Hypaque isolated mononuclear cells. The first method contains environmentally harmful reagents, and the second is laborious in the preanalytical steps. Cell preparation tubes (CPTs) are ready-to-use Ficoll Hypaque-based tubes that avoid the time-consuming and critical loading on Ficoll Hypaque. We examined whether CPTs can be used to determine melanoma cell dissemination in peripheral blood. We first investigated whether melanoma cells were retained in the mononuclear cell layer. All six morphologically different melanoma cell lines studied in the spiking experiments were retained in the upper layer. In further experiments, we were able to detect low dilutions of added SK MEL-28 cells more consistently after nested RT-PCR for tyrosinase or MART-1 in the RNA isolated from mononuclear cells from CPTs than from RNA isolated with the GITC method. In addition, RNA was extracted from paired blood samples from 24 analysable stage III and stage IV melanoma patients and analysed for the presence of tyrosinase and MART-1 RNA using both the CPT/RNeasy and the whole blood/GITC method. The quality of the CPT/RNeasy RNA was better than the RNA isolated from whole blood with GITC/phenol. However, the RT-PCR results were less unequivocal: MART-1 mRNA was more often detected with CPTIRNeasy compared with whole blood/GITC (six versus three), whereas tyrosinase mRNA was found less often in CPT/RNeasy RNA (two versus eight). Taken together these results suggest that the CPT isolation method is suitable for the isolation of mononuclear cells, including melanoma cells. PMID- 10803713 TI - Skin equivalent: an attractive model to evaluate early melanoma metastasis. AB - The growth patterns and morphological phenotype of four human melanoma cell lines with different metastatic potentials were investigated in submerged and in air exposed (skin equivalent) keratinocyte-melanoma cell co-cultures. In contrast to the submerged co-cultures, all four cell lines formed sharply demarcated tumour cell nests within the epidermal compartment of the skin equivalent model, with the morphology highly mimicking the in vivo situation. Differences among the melanoma cell lines tested were observed with respect to the number of clusters formed and the ability to exhibit invasive growth. Only the two metastatic cell lines were able to invade the dermal compartment. Screening of cellular adhesion molecules revealed that the expression patterns in different cell lines were heterogeneous and remained unchanged during the whole culture period, irrespective of whether the melanoma cells were located in the epidermal or dermal compartment. A correlation was found between expression of a lower number of different cellular adhesion molecules and the ability to acquire invasive growth capability. Our results indicate that melanoma cells exhibit a heterogeneous growth behaviour when co-cultured with human keratinocytes, and the air-exposed skin equivalent model was shown to be suitable for studying differences in growth patterns and potential invasive behaviour. PMID- 10803714 TI - Hypoluminescence microscopy of pigmented skin lesions. AB - Since the 'renaissance' of epiluminescence microscopy (ELM), histological correlation of ELM structures has been the subject of many investigations. Direct correlation is difficult because of the methodological differences between ELM and histopathology. In order to further understand the features of pigmented skin lesions (PSLs), we studied whether hypoluminescence microscopy (HLM) had any advantages over ELM. Twenty pigmented skin lesions scheduled for surgical excision were chosen randomly for this study. After excision, the lesions were studied using standard ELM and an HLM technique. For the latter, illumination was performed from the dermal side. The HLM pattern was clearly different from that of ELM. In all lesions the 'deeper' (dermal) structures became more visible. Some structures already visible in ELM appeared more visible, particularly structures apparently localized in deeper layers of the PSL. For highly pigmented lesions the difference in the two approaches was striking. With the HLM technique details became visible, and structures which were hardly or not visible with the classic ELM approach could be easily distinguished and evaluated. In conclusion, HLM is a simple, easy to use, reproducible ex vivo technique that is able to provide additional information to the conventional ELM technique. PMID- 10803715 TI - Melanoma and additional primary cancers. AB - The aim of this study was to examine the occurrence of additional, unrelated primary cancers in patients with melanoma. Data from the hospital-based, melanoma registry of a specialist unit (the Melanoma Unit at Charing Cross Hospital, London, UK) were compared with the incidence rates in a population-based cancer registry (the Thames Cancer Registry). In total, 2076 patients with the histological diagnosis of melanoma established between 1960 and 1997 who were registered with the Melanoma Unit at Charing Cross Hospital, were included in the study. Patterns in time and in the tumour type of the additional cancers were analysed in the cohort. The relative risk of subsequent cancers was evaluated, the number of expected cancers being calculated by applying incidence rates in the population of south-east England to the person-years of follow-up in the cohort. Sixty-six (3%) of the 2076 evaluable patients had a history of, or developed, 69 histologically verified additional cancers, the commonest being colorectal, breast and lymphoma. Twenty-six additional cancers preceded the diagnosis of melanoma by 1-42 years, 16 were diagnosed within 12 months and 27 followed the diagnosis of melanoma, 1-28 years later. Seven cancers occurred after chemotherapy for metastatic melanoma: two colorectal, one bladder, one renal, two myelodysplasias and one acute myeloid leukaemia. The relative risk of additional cancers developing after the diagnosis of melanoma during the 8537 evaluable years-at-risk was 0.64 (95% confidence interval 0.41-0.96; P = 0.0006). Thus, the risk of additional cancers following treatment for metastatic melanoma in this cohort is small. The potential influence of other factors on the occurrence of additional cancers observed overall in this study requires further investigation. PMID- 10803716 TI - Thick cutaneous malignant melanoma: a reappraisal of prognostic factors. AB - The prognosis of patients with thick (>3 mm) cutaneous malignant melanomas is generally poor; however, some cases survive far longer than expected. Thus tumour thickness cannot serve as the only predictor of disease course in the individual patient. The aims of the current study were to evaluate the clinical outcome of patients with thick (>3 mm) cutaneous melanoma and test the prognostic value of a series of clinicopathological parameters on disease-free and cause-specific survival. We retrospectively evaluated 140 patients with stage I cutaneous melanoma >3 mm in thickness. Disease-free and cause-specific survival rates (Kaplan-Meier method) were compared using the log rank test. A multivariate analysis (Cox proportional hazards model) was used to determine the independent effect of each variable on prognosis. The overall 5-year and 10-year disease-free survival rates were 35.5% and 29.3%, respectively, whereas the overall 5-year and 10-year cause-specific survival rates were 55.3% and 47.7%, respectively. In the univariate analysis, the following factors were found to be significantly associated with the disease-free and cause-specific survival: tumour thickness, mitotic rate/mm2, type of invasive front, ulceration, thickness of the nodular component and predominant cell type. In addition, the presence of vascular invasion was significantly correlated with the risk of metastases but not with survival. In the multivariate analysis (Cox proportional hazards model), only tumour thickness (both as a continuous variable and >7.5 mm), infiltrating invasive front, presence of ulceration and mitotic rate/mm2 (both as a continuous variable and >10 mitoses/mm2) were significant independent predictors of poorer clinical outcome. PMID- 10803717 TI - Prognostic impact of the type of anaesthesia used during the excision of primary cutaneous melanoma. AB - The prognostic value of the type of anaesthesia used for the excision of malignant tumours has been a subject of controversy. Cell-mediated as well as humoral immune responses can be compromised after general anaesthesia, and recurrences may therefore occur more frequently. On the other hand, excision of primary tumours under local anaesthesia might also influence the prognosis unfavourably. The aim of the present study was to determine the prognostic impact of general and local anaesthesia for the primary excision of cutaneous melanoma. Follow-up data of 4329 patients on the Central Melanoma Registry of the German Dermatological Society were analysed. Cox proportional hazards analysis was performed to evaluate the independent significance of the prognostic factors, and survival probabilities were calculated for matched pairs using Kaplan-Meier estimates. Statistical analysis revealed an independent significant effect on survival for tumour thickness, ulceration, level of invasion, anatomical site and gender. General anaesthesia for primary excision of melanoma was associated with a decrease in the survival rate (relative risk 1.46, P<0.0001). This study revealed a slight but significantly increased risk of death for patients treated with general anaesthesia for the primary excision of melanoma. Therefore local anaesthesia should be preferred for the treatment of primary melanoma. PMID- 10803718 TI - A phase II study of biochemotherapy for the treatment of metastatic malignant melanoma. AB - The use of interleukin-2 (IL-2) and interferon-alpha (IFNalpha) in combination with chemotherapy for the treatment of advanced malignant melanoma has generated considerable interest. In particular, the relatively high number of durable complete responses has suggested this may be a significant advance in the treatment of malignant melanoma. We report our experience at the University of Colorado in 43 patients, including many with poor prognostic factors. Patients received cisplatin 20 mg/m2 on days 1-4, vinblastine 1.6 mg/m2 on days 1-4, dacarbazine 800 mg/m2 on day 1, IL-2 9 x 10(6) IU/m2 per day intravenously over 24h on days 1-4 and IFNalpha 5 x 10(6) IU/m2 per day subcutaneously on days 1-5 every 3 weeks. The median follow-up for all patients was 34 months. Responses were seen in 20 patients (47%, 95% confidence interval 31-62%) and comprised five complete responses (CRs) (12%) and 15 partial responses (PRs) (35%). Two patients achieving a CR remain disease free at 45 and 47 months follow-up. In addition three patients who obtained a surgical CR and another with only minor residual changes on computed tomography scan have not progressed at 27, 30, 40 and 27 months, respectively. Toxicity was manageable, but all patients had at least one grade 3 or 4 toxicity, predominantly hypotension and neutropenia. There were no treatment-related deaths. In conclusion, the response rate and duration is within the range previously reported for biochemotherapy. The results of ongoing randomized studies are awaited to better define the value of biochemotherapy in the treatment of advanced melanoma. PMID- 10803719 TI - Early detection of melanoma: an educational campaign in Padova, Italy. AB - To evaluate a public campaign for the early referral and treatment of cutaneous melanoma, an educational programme based on self-selection by subjects was organized in Padova, Italy in 1991. In the period from 1991 to 1996, 90,000 leaflets containing information on naevi, melanoma and skin self-examination were mailed to each household, reaching a population of 243,000 subjects. A total of 2050 individuals requested a skin check as a result of the leaflet. Most were at low risk, the majority being female (68%) and aged under 40 years (51.6%), with no risk factors (58.3%). One hundred and ninety subjects were referred for surgery for pigmented and non-pigmented suspect lesions. Histological diagnoses, obtained for all lesions, comprised 13 melanomas, 17 dysplastic naevi, 17 basocellular carcinomas, 140 pigmented benign lesions and three lesions of other types. The percentage of thin melanomas (< 1.50 mm) was 92.3%. Three hundred and fifty patients considered at risk at the first skin examination attended regular follow-up examinations. The sensitivity and predictive positive value of the visual examination were 92.8% and 6.8%, respectively. The impact of this campaign was evaluated in the Local Health District of Padova, comparing data from the pre campaign period (1987-1990) with those from the campaign period (1991-1996); a trend towards a lower stage was observed (mean thickness 2.0 mm versus 1.50 mm; P < 0.02). PMID- 10803720 TI - Combined treatment with thymosin-alpha1 and low dose interferon-alpha after dacarbazine in advanced melanoma. PMID- 10803721 TI - Estimation following extension of a study on the basis of conditional power. AB - Proschan and Hunsberger (1) propose a method based on conditional power for designed extension of a study beyond its originally intended sample size. Their data-dependent sampling method can be viewed as a two-stage procedure in which the target total sample size is dependent upon the data observed at the first stage. We demonstrate that the maximum likelihood estimate of the parameter of interest upon completion may be biased, and that this bias is similar in direction and magnitude to that commonly associated with estimation following a group sequential test with predetermined target total sample size. Furthermore, we show how a bias adjusted estimate may be formed. PMID- 10803722 TI - Center-weighting issues in multicenter clinical trials. AB - For linear model analysis of data from multicenter clinical trials, results are presented to assist in comparing commonly used approaches in which individual center results are weighted either equally or according to the amount of information they provide. A measure of center size imbalance directly relating to the relative efficiencies of the two weightings is defined. It is demonstrated that when centers of different sizes are given equal weight, the method by which sample size is conventionally calculated is incorrect. Center weighting resulting from use of random-effect models is also discussed. PMID- 10803723 TI - Multiple imputation compared with some informative dropout procedures in the estimation and comparison of rates of change in longitudinal clinical trials with dropouts. AB - Statistical analysis based on multiple imputation (MI) of missing data when analyzing data with missing observations is gaining popularity among statisticians because of availability of computing softwares; it might be tempting to use MI whenever data is missing. An important assumption behind MI is the "ignorability of missingness." In this paper, we demonstrate the use of MI in conjunction with random effects models and several other methods that are devised to handle nonignorable missingness (informative dropouts). We then compare the results to assess sensitivity to underlying assumptions. Our focus is primarily to estimate and compare rates of change (of a primary variable). The application dataset has a high dropout rate and has features to suggest informativeness of the dropout process. The estimates obtained under random effects modeling with multiple imputation were found to differ substantially from those obtained by methods devised to handle informative dropouts. PMID- 10803724 TI - ROC surface: a generalization of ROC curve analysis. AB - Receiver operating characteristic (ROC) curve analysis is widely used in biomedical research to assess the performance of diagnostic tests. Much of the work has been directed at developing accurate indices to describe ROC curves and appropriate statistics to test differences between them. The analysis, however, is largely built on the assumption that the test results are dichotomous. We generalize the ROC curve analysis to allow for tests to have more than two outcomes. The generalized ROC curve constitutes a surface. We propose to use the volume under the surface to measure the accuracy of a diagnostic test. PMID- 10803725 TI - Design and estimation in the limiting dilution assay when concentrations vary widely among samples. AB - Limiting dilution assays (LDA) are used to estimate an unknown cell fraction of interest within a sample. This paper discusses a method for designing an LDA using the distribution of the cell fraction of interest, examining three different design approaches: geometric progression, equally spaced log, and equiprobability. Two common estimation methods, minimum chi-square and maximum likelihood, also are investigated. These designs and estimation methods, coupled with varying numbers of wells per dilution and dilutions per design, are compared quantitatively through computer simulation. Performance measures computed were mean relative bias and mean squared error. PMID- 10803726 TI - Comparison of bracketing and matrixing designs for a two-year stability study. AB - In the U.S. Food and Drug Administration (FDA) guidelines for stability testing of new drug products, both bracketing and matrixing designs were suggested as the statistical designs. More recently, they have increasing attention from pharmaceutical companies, because both designs reduce the cost of stability studies. The purpose of this paper is to investigate both designs in terms of the power of detection of significant difference between slopes, and use the mean square error to evaluate the precision of estimated drug shelf life. Additionally, the distributions of both designs are compared by using 1000 simulations. PMID- 10803727 TI - Assessment of similarity between dissolution profiles. AB - In vitro dissolution equivalence has been assessed through profile comparison between the test and reference formulations for postapproval changes. Functions of the absolute mean difference and average squared mean differences were two of the often-used criteria to evaluate distance or similarity between general profiles. The two functions and their method of moment estimators are studied in this paper as an application to similarity assessment of two dissolution profile. Due to the complexity of the distributions of these estimators, the confidence intervals obtained from the bootstrap method were used for testing the hypothesis of dissolution similarity. The size and power of two procedures are examined with a simulation study. A numerical example is used to illustrate the application of the procedures. PMID- 10803728 TI - A robust summarize-regress procedure for tissue-specific pharmacokinetics. AB - The destructive sampling necessary to obtain tissue-specific pharmacokinetics of radiolabeled ligands does not permit true time-dependent models to be estimated. To adjust for this, small groups of animals are sacrificed at varying time points and the percent injected dose per gram (PID) of radiolabeled ligands is summarized for each tissue of interest. Usually the average PID at each time point is modeled to obtain estimates of pharmacokinetic parameters for each tissue when utilizing summarize-regress procedures. In this paper, it is recommended that the median should be the summary measure at each time point to be modeled in these circumstances. The discrepancy that exists between modeling the average and median PID is demonstrated by an actual experiment. Finally, simulation studies are performed and the results are compared when using these two summary measures. PMID- 10803729 TI - Issues in use of SAS PROC.MIXED to test the significance of treatment effects in controlled clinical trials. AB - A project that originated with the aim of documenting the implications of dropouts for tests of significance based on general linear mixed model procedures resulted in recognition of problems in the use of SAS PROC.MIXED for this purpose. In responding to suggestions and criticisms, we have further analyzed simulated clinical trial data with realistic autoregressive structure, using alternative error model formulations, different approaches to the use of covariates to model dropout patterns, and different ways to include the critical time variable in the mixed model. Results emphasize the sensitivity of the PROC.MIXED tests of significance for GROUP and TIME x GROUP equal slopes hypothesis to less than optimal modeling of the error covariance structure. Even with the authoritatively recommended best available modeling of the error structure, model formulations that made use of the REPEATED statement did not maintain conservative test sizes when covariates were required to model dropout data patterns. Random coefficients models that employed the RANDOM statement did permit appropriate covariate controls, but the tests of significance for treatment effects were lacking in power. After examining a variety of alternative PROC.MIXED model formulations, it is concluded that none provided both Type I error protection and power comparable to that of simple two-stage analysis of covariance (ANCOVA) procedures for confirming the presence of true treatment effects in controlled clinical trials. Other issues examined in this article concern treating baseline scores as both covariate and initial repeated measurement to which a linear means model is fitted, failure to take advantage of the regression of repeated measurements on time in modeling time as an unordered categorical variable, and fitting linear regression models to nonlinear response patterns. PMID- 10803730 TI - Renal dopaminergic mechanisms and hypertension: a chronology of advances. AB - Dopamine (DA) has been shown to influence kidney function through endogenous synthesis and subsequent interaction with locally expressed dopamine receptor subtypes (D1, D5 as D1-like and D2, D3, and D4 as D2-like). DA, and DA-receptor specific agonists and antagonists can alter renal water and electrolyte excretion along with renin release when infused systemically or intrarenally. Such effects are brought about by a combination of renal hemodynamic and direct tubular effects evoked along the full length of the nephron. The cellular mechanisms that direct these dopamine-mediated renal electrolyte fluxes have recently been clarified and include alterations in adenylyl cyclase, phospholipase C, and phospholipase A1 activity. The dopaminergic system also interacts directly with the renal kallikrein-kinin, prostaglandin and other neurohumoral systems. Aberrant renal dopamine production and/or dopamine receptor function have been reported in salt-dependent and low-renin forms of human primary hypertension as well as in genetic models of animal hypertension, including the SHR and Dahl SS rat. DA D1 or D3 receptor knockout mice have been shown to develop hypertension. PMID- 10803731 TI - Renal dopaminergic mechanisms in renal parenchymal diseases, hypertension, and heart failure. AB - The recovery of renal function in renal transplant recipients is accompanied by an enhanced ability to synthesize dopamine (DA), which may contribute to maintain sodium homeostasis. Patients suffering from chronic renal parenchymal disease, a well-recognized form of salt sensitive (SS) hypertension, have a reduced ability to produce DA that correlates well with deterioration of renal function. In patients afflicted with IgA nephropathy, but normal renal function, urinary excretion of DA correlated positively with BP responses to changes from 200 to 20 mmol/day salt intake. In black salt resistant (SR) normotensives (NT) and SR hypertensives, under low salt intake (40 mmol/day), but not SS-NT and SS-HT, the saline infusion induced increments of DA and DOPAC urinary excretion correlated significantly with increments of sodium urinary excretion and sodium fractional excretion. Patients afflicted with heart failure (HF) have a reduced delivery of L-DOPA to the kidney, accompanied by an increase in DA/L-DOPA urinary ratios. This suggests that HF patients have an increased ability to take up or decarboxylate L-DOPA. Sodium restriction resulted in a significant decrease in urinary L-DOPA, DA and DOPAC in HF patients, suggesting that the system responds to sodium. It is concluded that activity of renal dopaminergic system may be altered in SS subjects, despite the level of their BP, and an enhanced delivery of L-DOPA to the kidney may be beneficial in edema formation states. PMID- 10803732 TI - Dopamine in heart failure and critical care. AB - Dopamine is widely used in critical care to prevent renal function loss. Nevertheless sufficient evidence is still lacking of reduction in end points like mortality or renal replacement therapy. Dopaminergic treatment in chronic heart failure (CHF) has provided an example of unexpected adverse outcome. Pharmacoepidemiological data. Provide additional evidence, finding excess mortality in current ibopamine users (relative risk 2.03 in NYHA I-II CHF, 1.37 in NYHA III-IV), while no relation was found with antiarrhythmic use. In critical care, studies after infrarenal aortic surgery or during septic shock, respectively, failed to find, expected specific renal effects of dopamine. Effects on splanchnic flow mainly depend on baseline flow levels. The implications of recently documented unwanted effects of dopamine, like reduced ventilation and oxygenation during hypoxia, are discussed. In conclusion, controlled clinical trials remain mandatory to assess the overall clinical effects of dopamine in critical care. PMID- 10803733 TI - Localization of dopamine receptor subtypes in systemic arteries. AB - Dopamine D1-D5 receptor protein immunoreactivity was investigated in different sized pial, renal and mesenteric artery branches using immunohistochemical techniques and anti-dopamine D1-D5 receptor protein antibodies. Faint dopamine D1 receptor protein immunoreactivity was observed in smooth muscle of tunica media of pial, renal and mesenteric artery branches. Dopamine D2 receptor protein immunoreactivity was located in the adventitia and adventitia-media border of pial and renal artery branches and to a lesser extent of mesenteric artery branches. No dopamine D3 receptor protein immunoreactivity was observed in pial and mesenteric arteries. In renal arteries a moderate dopamine D3 receptor immunoreactivity was detectable in the adventitia and adventitia-media border. A strong dopamine D4 receptor protein immunoreactivity displaying the same localization of dopamine D2 receptor protein was observed in pial and mesenteric arteries, but not in renal artery branches. Moderate dopamine D5 receptor protein immunoreactivity was observed in smooth muscle of the tunica media of pial, renal and mesenteric artery branches. Bilateral removal of superior cervical ganglia, from which sympathetic supply to cerebral circulation originate abolished dopamine D2 and D4 receptor protein immunoreactivity in pial arteries but was without effect on dopamine D1 and D5 receptor protein immunoreactivity. These findings indicate that systemic arteries express dopamine D1-like (D1 and D5) and D2-like (D2, D3 and D4) receptor subtypes displaying respectively a muscular (postjunctional) and prejunctional localization. The specific distribution of dopamine D2-like receptor subtypes in systemic arteries suggests that they may have a different role in regulating blood flow through the vascular beds investigated. PMID- 10803735 TI - Mechanisms by which intrarenal dopamine and ANP interact to regulate sodium metabolism. AB - Maintenance of a normal blood pressure requires a precise and fine-tuned regulation of salt metabolism. This is accomplished by a bidirectional regulation of renal tubular sodium transporters by natriuretic and antinatriuretic hormones. Dopamine, produced in the renal proximal tubular cells, plays an important role in this interactive system. Dopamine inhibits the activity of Na+,K+ ATPase as well as of many important sodium influx pathways in the nephron. These effects of dopamine are particularly pronounced in situation of sodium loading. There is an abundance of evidence suggesting that the natriuretic effects of ANP are to a large extent mediated via renal dopamine 1 like receptors. The renal tubular dopamine 1 like receptors are, under basal conditions, mainly located intracellularly. ANP and its second messenger, cGMP, cause a rapid translocation of the dopamine 1 like receptors to the plasma membrane. This phenomenon may explain how ANP and dopamine act in concert to regulate sodium metabolism. Regulation of sodium metabolism and blood pressure is critically dependent on a normal function of the renal dopamine system. Hence, abnormalities in the interaction between dopamine and ANP may predispose to hypertension. PMID- 10803734 TI - Biphasic effects of dopamine on 86rubidium uptake in rat renal proximal tubules. AB - The mechanism(s) by which dopamine inhibits Na+-K+-ATPase activity in the renal proximal tubule is still controversial. We studied the short-term effects of dopamine on the sodium pump in rat renal proximal tubule suspensions with the 86Rb uptake method. Dopamine and the D1-like agonist, SKF81297, initially stimulated Na+-K+-ATPase activity at 5 min and subsequently inhibited it at 10 min and 20 min; the inhibition by 10 microM dopamine at 20 min was 21.3 +/- 4.5%. The inhibitory effect of dopamine on Na+-K+-ATPase activity was mimicked by thymeleatoxin (a classical protein kinase C [PKC] agonist) while Sp-8-CPT-cAMPS (a protein kinase A [PKA] agonist) had no effect. However, the combination of the PKC and PKA agonists mimicked the biphasic effects of dopamine and SKF81297. Rp-8 CPT-cAMPS (a PKA inhibitor), U-73122 (a phospholipase C inhibitor), or calphostin C (a PKC inhibitor), blocked the dopamine-mediated biphasic effects on Na+-K+ ATPase activity. It is suggested that the biphasic effects of dopamine on Na+-K+ ATPase activity (an initial stimulation and a subsequent inhibition) are transduced by activating both PKA and PKC through a D1-like receptor. PMID- 10803736 TI - The role of intrarenal nitric oxide in the natriuretic response to dopamine receptor activation. AB - Dopamine and dopamine-1 receptor agonists produce diuresis and natriuresis by causing changes in renal hemodynamics and by the activation of dopamine-1 receptors located within the various regions of the nephron. Nitric oxide plays an important role in the maintenance of systemic and regional hemodynamics. The present study was undertaken to investigate the effect of locally generated nitric oxide on renal function and its potential influence on the renal responses to dopamine-1 receptor agonists. The intrarenal infusion of a nitric oxide synthase inhibitor, L-NAME, (50 microg/kg min for 90 min) in anesthetized rats produced significant decreases in urine volume, urinary sodium excretion, glomerular filtration rate and fractional sodium excretion. These changes in renal function were associated with a concomitant decrease in urinary nitrate excretion, an indicator of nitric oxide release. However, L-NAME at this dose did not produce any significant changes in mean arterial pressure or heart rate. Intravenous infusion of fenoldopam (1 microg/kg min for 30 min), a selective dopamine-1 receptor agonist, produced diuresis and natriuresis without causing any changes in mean arterial pressure and heart rate. These renal effects of fenoldopam were significantly attenuated in animals that received the simultaneous infusion of L-NAME (intrarenal). Similar results were obtained with dopamine in that the natriuretic and diuretic response to dopamine was also attenuated during simultaneous infusion of dopamine with L-NAME. In addition, the diuresis and natriuresis produced by fenoldopam and dopamine was associated with increases in urinary nitrate excretion. Interestingly, these increases in the nitrate levels seen with fenoldopam and dopamine were also significantly reduced in the presence of L-NAME. These results indicate that intrarenal nitric oxide plays an important role in regulating renal sodium excretion and that an intact renal nitric oxide system is required for the full expression of diuretic and natriuretic response seen during dopamine-1 receptor activation. PMID- 10803737 TI - Natural killer cells as potential tools in melanoma metastatic spread control. AB - A large basic knowledge is now available on natural killer (NK) cell biology in mice and humans. Intensive research during the 1990s has clarified many aspects on the specificity and receptors of NK cells. It is now possible to apply this knowledge to the study of NK cell responses in important medical problems. The general aim of this commentary is to focus the attention of clinical and basic immunologists and oncologists on the possible involvement of NK cells in new biologically oriented antitumor protocols. Here we comment on the more recent studies of NK cell recognition of melanoma cells, with particular emphasis on the NK-activating and -inhibiting receptors and their ligands regulating NK cytotoxicity. PMID- 10803738 TI - Mechanisms involved in the potentiation of melphalan by the bioreductive compound THNLA-1 in vitro. AB - 9-[3-(2-Nitro-1-imidazolyl)propylamino]-1,2,3,4-tetrahydroacridine hydrochloride (THNLA-1) is a 2-nitroimidazole-based, weakly DNA-intercalating bioreductive agent that significantly potentiates the toxic effects of commonly used antitumor drugs such as melphalan (L-PAM) or cis-DDP in sensitive or resistant cell lines in culture, as well as in solid tumors in mice. Potentiation in vitro was observed when cells were preexposed to THNLA-1 under hypoxic conditions before exposure to L-PAM under aerobic conditions. In this study we investigated possible mechanisms involved in the potentiation of L-PAM by THNLA-1 in V79 Chinese hamster cells. Limited depletion of glutathione with buthionine sulfoximine or THNLA-1 under hypoxic pretreatment conditions accounted for only 8.3% of the potentiation induced by THNLA-1. However, DNA, RNA, and protein synthesis were inhibited in a synergistic way in cells preexposed to THNLA-1 under hypoxic conditions (2 h, 37 degrees C) and then coexposed to various doses of L-PAM under aerobic conditions (1 h, 37 degrees C). Cell cycle analysis by flow cytometry showed a slow traverse through the S phase in the L-PAM-alone treated cells. However, this phenomenon was more prominent in the THNLA-1 plus L PAM-treated cells. Under aerobic co-incubation conditions with L-PAM, no difference was observed in the cell cycle of L-PAM-alone-treated cells vs. THNLA 1 plus L-PAM-treated cells. Significantly increased apoptosis was observed in the hypoxia-pretreated cells with THNLA-1, 12 and 24 h posttreatment. Comet and alkaline elution assay analysis showed increased DNA cross-links in the hypoxia pretreated cells with THNLA-1 compared to the L-PAM-alone-treated cells. Finally, potential lethal damage repair was totally suppressed only in the hypoxia pretreated cells with THNLA-1. In conclusion, DNA damage and hindrance in its repair are the most important mechanisms in the potentiation of L-PAM by THNLA-1, under hypoxic pretreatment conditions. PMID- 10803739 TI - Morphologic aspect of the placenta in young and adult pregnant rats bearing Walker 256 carcinoma. AB - In the present study we investigated the influence of Walker 256 tumor growth on the modification of placental morphology and on fetal development in young and adult pregnant rats. After mating, female rats were divided into six groups: young control pregnant (Y), young pregnant with tumor (Yw), young pregnant injected with ascitic fluid (Ya), adult control pregnant (A), adult pregnant with tumor (Aw), and adult pregnant injected with ascitic fluid (Aa). Rats from tumor bearing groups (Yw and Aw) were injected with 2.5 x 10(6) viable tumor cells into the right flank. Rats from Ya and Aa groups received daily inoculations of ascitic fluid (2.0 ml, i.p.) obtained from tumor-bearing rats without tumor cells. After 21 days, all animals were killed and the placentas were weighed and fixed with paraformaldehyde for histological analysis. Compared with control groups (Y and A), both tumor-bearing groups (Yw and Aw) presented the following changes: i) hemorrhage in the decidua and in the trophoblast giant cell layer; ii) disarrangement of the spongy zone, iii) restricted delimitation of the maternal and fetal blood vessels in the placental labyrinth; iv) hemorrhage and edema in the placental labyrinth. Similar results were observed in the placenta of groups injected with ascitic fluid (Ya and Aa). These results indicate that tumor development during pregnancy can have deleterious effects on placenta and fetus. These observations extend our previous data of extensive fetal reabsorption in both pregnant tumor-bearing and ascitic fluid-injected animals. These changes in placental morphology may be related to the synthesis and release of some factors by the tumor and the host cells, which could act directly or indirectly on placental tissue. PMID- 10803740 TI - Comparison of flow cytometry and RT-PCR for the detection of ovarian cancer cells in peripheral blood. AB - Recently, there has been significant effort in developing techniques designed to detect disseminated tumor cells in the peripheral blood (PB). These techniques include immunocytochemical staining of cytocentrifuge slides, flow cytometry, and RT-PCR. Several authors reported various results concerning the sensitivity of the detection limit when applying these methods. The aim of this study was to assess the value of two methods in the detection of ovarian carcinoma cells in the PB. For tumor cell detection we compared RT-PCR to immunomagnetic enrichment followed by flow cytometric analysis. In a model system, single cell suspensions of ovarian cancer cell lines were mixed with full blood samples from healthy donors in order to determine the sensitivity limit of the two methods and to analyze the reproducibility of each. In a multiparameter flow cytometric analysis, tumor cells were defined as cytokeratin 7/8 positive and CD45 negative. RNA was screened for MUC1 mRNA by RT-PCR. MUC1 mRNA expression turned out not to be a specific marker of disseminated ovarian cancer cells, because a weak expression was also found in samples of healthy persons. Using immunomagnetic enrichment followed by flow cytometry, one carcinoma cell per 1 x 10(6) leukocytes was detectable. However, a minimum of 10 ml blood had to be analyzed in order to clearly distinguish real positive tumor cells from false-positive signals. PMID- 10803741 TI - Influence of the fluoroquinolone ofloxacin on the intrinsic expression of multidrug resistance phenotype in HCT-8 human colon carcinoma cells. AB - The influence of antibiotics, particularly ofloxacin (OF), a commonly used antimicrobial fluoroquinolone, on the multidrug resistance (MDR) phenotype of the HCT-8 cell line was studied. This cell line was grown in OF containing medium for several months and the expression of the MDR phenotype was followed through the analysis of the expression and functionality of the P-glycoprotein (Pgp), the chemosensitivity to daunorubicin (DNR), and the mRNA expression of mdr-1, multidrug resistance-associated protein (MRP), and topoisomerase IIalpha and IIbeta genes. Replacement of OF by penicillin streptomycin (PS) resulted in a significant decrease in mdr-1 mRNA expression, which was found to correlate with a decrease in the expression and functionality of the Pgp. After antibiotic starvation for 4 weeks, cells grown in antibiotic-free medium were then exposed to PS or OF; these cells showed an increase in mdr-1 mRNA/Pgp and MRP mRNA expression without a decrease in DNR cytotoxicity. OF cultured cells exhibited a significant increase in Pgp expression without evidence of the functionality of the Pgp. An increase in topoisomerase IIalpha mRNA expression was observed with time and with the number of passages of the cell line without any relationship to the presence of antibiotics in the culture medium. These results showed that extensive use of antibiotics, particularly the quinolones, can modify the phenotype of the HCT-8 colon adenocarcinoma cell line. PMID- 10803742 TI - Combination studies of antifolates with 5-fluorouracil in colon cancer cell lines. AB - The combined cytotoxic effects of the thymidylate synthase (TS) inhibitors 5 fluorouracil (5FU) and different antifolates were studied in seven colon cancer cell lines. Growth inhibition of the antifolates, Nolatrexed, Raltitrexed, GW1843U89, or MTA in combination with 5FU, was determined and multiple drug effect analysis showed that the drugs acted mostly additively. The only synergistic interaction was found for 5FU and Nolatrexed in the LS174T cell line. Also Raltitrexed and 5FU were slightly synergistic in WiDr/F cells grown at low folate levels, but for the other cell lines grown at high folate levels this combination was more antagonistic. GW1843U89 and 5FU were mainly additive, while 5FU and MTA showed antagonism in WiDr and additivity in LS174T. The effect of the drugs at their target was evaluated by in situ TS inhibition. We observed lower TS activity in all cells when two drugs were used instead of one. Statistical analysis revealed that none of the values of the combinations was higher or lower than could be expected from the product of the effect of single drugs. We concluded that the effects on TS inhibition were additive for all 5FU/antifolate combinations in all cell lines. DNA strand break formation, as a result of TS inhibition, was measured by means of a fluorometric analysis of DNA unwinding. Raltitrexed-induced DNA damage was significantly increased by 5FU in WiDr cells [single agent: 67% double stranded (ds) DNA, combination: 39% ds DNA, P<0.0001]. In LS174T a trend for antagonistic effects was observed for combinations of MTA, GW1843U89, or Raltitrexed and 5FU. The combinations showed additive effects in WiDr/F cells. The overall conclusion of the three assays in each of the cell lines indicated that 5FU and antifolate combinations were predominantly additive in colon cancer cells. PMID- 10803743 TI - Are the biologic and clinical effects of the COX-2-specific inhibitors an advance compared with the effects of traditional NSAIDs? AB - This has been an unusual year for the accumulation of evidence regarding the clinical effects of inhibition of cyclooxygenase (COX)-1 and COX-2. This article reviews the available data regarding the clinical effects of the new COX-2 specific inhibitors, and speculates about the importance of the data as they relate to the treatment of patients with chronic pain and/or inflammation. PMID- 10803744 TI - Corticosteroids in the treatment of rheumatologic diseases. AB - The year 2000 marks the fiftieth anniversary of the awarding of the Nobel Prize for Medicine to Hench, Kendall, and Reichstein for their discovery of glucocorticoid treatment of rheumatic diseases. The efficacy and toxicity of glucocorticoids has remained a matter of contention ever since, with debate continuing over their place in the therapeutic armamentarium of rheumatologists. Few if any rheumatologists would not prescribe glucocorticoids, however, and review of new data on their use, efficacy, and toxicity remains topical. Perceived advances in the ability to manage osteoporosis arising from glucocorticoid use has allowed focus to shift onto other toxicities, including vascular disease, but important advances in our understanding of the mechanism of action of glucocorticoids are still lacking. PMID- 10803745 TI - Rheumatoid arthritis. PMID- 10803746 TI - Are T cells in rheumatoid synovium aggressors or bystanders? AB - T cells have been directly associated with rheumatoid arthritis (RA) because they represent the largest cell population infiltrating the synovium. Their direct contribution to disease and joint destruction has been more difficult to demonstrate. Locally, they interact with other blood-derived and resident cells. Some T cells may contribute to disease through the secretion of cytokines. Indeed, interleukin-17, a T-cell-specific cytokine, is produced by RA synovium and acts as a bone and cartilage destructive factor. In addition, it increases the production of proinflammatory cytokines by monocytes and further enhances their effects on matrix destruction. Once considered bystanders in RA, T cells can now be classified as aggressors through their direct and indirect contribution to destruction. In particular, a subset of Th1 T cells can aggravate the proinflammatory and destructive pattern associated with monocyte activation. Manipulation of this subset may control the destructive pattern. Such a result can be achieved when a switch can be induced from a destructive pattern to a protective one leading to repair. PMID- 10803747 TI - Activation of synoviocytes. AB - The evaluation of molecular pathways has revealed various novel insights into rheumatoid arthritis pathophysiology during the past year. In addition, there is an increasing tendency toward analysis not merely of a single mechanism but rather of data addressing a substantial part of the cascade of events leading to cellular activation. Because synovial fibroblasts are key cells involved in joint destruction, this review outlines the events that trigger or inhibit the crucial pathways leading to their aggressive behavior. Major topics include cellular and humoral interactions (frequently modulated by cytokines), intracellular signaling and upregulation of gene transcription, and the deleterious effects on articular homeostasis. PMID- 10803748 TI - Pathogenesis of bone erosions in rheumatoid arthritis. AB - Patients with rheumatoid arthritis are at risk for the development of a generalized form of bone loss affecting the axial and appendicular skeleton. In addition, juxta-articular osteopenia and focal erosion of marginal and subchondral bone are commonly seen. The pathogenesis of focal bone erosions is an area of active investigation. Studies of tissue sections from sites of bone erosion in rheumatoid arthritis and in animal models of inflammatory arthritis have identified multinucleated cells with the phenotype of osteoclasts in bone resorption lacunae in these sites, suggesting that osteoclasts mediate a component of this pathologic bone loss. Numerous soluble and cell-membrane factors produced by rheumatoid synovial tissues are likely to play a role in the initiation and progression of bone erosions. In addition, recent studies suggest a role for T lymphocytes and their products in osteoclast-mediated bone loss. This paper reviews the cellular mechanisms and factors implicated in bone erosions in rheumatoid arthritis, and discusses the possible therapeutic strategies suggested by these findings. PMID- 10803749 TI - Markers of disease in rheumatoid arthritis. AB - Substantial progress can be noted in the efforts to demonstrate the usefulness of tissue-related markers of disease in rheumatoid arthritis and other joint diseases. The most informative studies use longitudinal analyses of well characterized patient groups. Emphasis should be on searching for markers which can be of prognostic significance. New markers need to be assessed in relation to existing ones, such as C-polysaccharide reacting protein and erythrocyte sedimentation rate, which, although not specific, are hard to beat as measures of inflammation. A newly identified matrix component, cartilage intermediate layer protein, has features which make it attractive as a potential cartilage specific marker. Many markers may not in the end prove clinically useful. They will, however, give important insight into pathogenic processes, and may help in evaluating new therapy. Finally, markers originally identified in humans have now proven their value in experimental arthritis. PMID- 10803750 TI - Gene transfer: from concept to therapy. AB - In the last few years, several novel strategies have been proposed for the treatment of rheumatoid arthritis (RA). Among them, gene therapy is considered a promising concept bearing the potential of highly specific targeting of relevant pathomechanisms. Early studies using gene transfer focused mainly on studying disease mechanisms, whereas recent research has put potential clinical applications to the forefront of attention. This has provided new answers to the question of how to deliver genes into the rheumatoid synovium as well as which pathways to target. Thus, significant progress has been made in the continued development of viral systems, including retro- and adenoviruses, as well as in the exploration of novel tools such as herpes virus-based systems or liposomes in combination with viral fusion proteins. When potential targets for gene transfer in RA are considered, two strategies have emerged: the first focuses on the delivery of secreted proteins, mainly cytokines and cytokine receptors, to inhibit inflammation in arthritic joints. Based on our growing knowledge about the pathogenesis of RA, however, there has also been substantial progress in exploring approaches that aim at interfering specifically with signaling pathways involved in the activation and apoptosis of synovial cells. The data from recent studies indicate the ability to selectively target specific disease processes by the differential expression of therapeutic genes in varying cell types and at different stages of disease, thus demonstrating the potential of gene transfer as an arthritis therapy. PMID- 10803751 TI - Gout update: from lab to the clinic and back. AB - Reviews and opinion articles stressing the need for better reproducibility of crystal identification in synovial fluid continue to be published. The possibility of definitive diagnosis of gout by crystal identification during intercritical periods appears now established. The problems associated with the use of allopurinol have also received some attention, as well as the strategies of dealing with allopurinol hypersensitivity. Gout in transplanted patients is reviewed, with special attention to the difficulties related to its treatment; various therapeutic options for this group are reviewed. The use of benzbromarone to reduce uricemia appears to be especially appropriate for this group of patients, in whom allopurinol is problematic. Finally, the pathogenetic mechanisms of the disease, and the relation between urate crystals and inflammation are reviewed. PMID- 10803752 TI - Formation of calcium pyrophosphate crystals: biologic implications. AB - The formation of calcium pyrophosphate dihydrate (CPPD) crystals in articular cartilage marks the earliest known phase of CPPD deposition disease. Although the exact mechanisms through which these crystals form remains unknown, work over the last year has added useful details to our current paradigms of crystal nucleation and growth. Key advances include (1) progress in understanding pyrophosphate elaboration and its modifiers, (2) further characterization of the enzymes responsible for pyrophosphate elaboration, and (3) the discovery of an association between two seemingly unrelated metabolic risk factors for CPPD deposition disease. PMID- 10803753 TI - Calcium crystal effects on the cells of the joint: implications for pathogenesis of disease. AB - In the past three years, there has been considerable progress in delineating the mechanism of calcium-containing crystal-induced cell activation: (1) the identification of Ca2+ influx and p42/44 mitogen-activated protein kinase activation as the signal transduction pathways; (2) induction of nuclear transcription factors of cyclic adenosine monophosphate (cAMP) response element binding protein, activator protein-1, and nuclear factor kappaB; (3) the differential role of crystal endocytosis and dissolution in crystal-induced metalloproteinase synthesis and mitogenesis; (4) crystal upregulation of matrix metalloproteinases, including MMP-13 but downregulation of tissue inhibitor of metalloproteinase-1 and -2, thus magnifying the degenerative effect of crystals. Phosphocitrate, a specific inhibitor of biologic effect of calcium crystals, reverses the degenerative effects of crystals. PMID- 10803755 TI - Bibliography. Current world literature. Clinical therapeutics. PMID- 10803754 TI - What is new about crystals other than monosodium urate? AB - Recently, attention has focused on the effects of weather conditions and seasonal changes on the incidence of acute microcrystalline events. Acute gout attacks are more frequent during the spring, but seasonal variations in the incidence of acute pseudogout attacks are less clearly defined. Genetic analysis of two unrelated families with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease showed linkage to the short arm of chromosome 5p. Several recent reports show CPPD crystal disease occurring in association with Gitelman syndrome, the hypocalciuric-hypomagnesemic variant of Bartter syndrome. Two signaling pathways, protein kinase C and adenyl cyclase, modulate generation of extracellular pyrophosphate by porcine cartilage chrondrocytes. These transduction mechanisms may provide potential targets for the treatment of CPPD crystal deposition disease. A controlled clinical trial showed ultrasound therapy to be beneficial in the treatment of symptomatic chronic calcific tendinitis of the shoulder. There is evidence that apatite crystals may contribute to cartilage damage in osteoarthritis and that therapeutic interventions to prevent the formation and biologic effects of the crystals may potentially retard the progression of the osteoarthritic process. PMID- 10803756 TI - Bibliography. Current world literature. Rheumatoid arthritis. PMID- 10803757 TI - Bibliography. Current world literature. Crystal deposition diseases. PMID- 10803758 TI - The role of ginsenoside-Rd in cisplatin-induced acute renal failure. AB - Ginsenoside-Rd has been proved to decrease the severity of renal injury induced by cisplatin, in which proximal urinaferous tubules represent the main site of injury. When ginsenoside-Rd was given orally at a dose of 1 or 5 mg/kg body weight/day for 30 consecutive days prior to cisplatin injection, the activities of the antioxidation enzymes superoxide dismutase and catalase were higher, while malondialdehyde levels in serum and renal tissue were lower in the treated rats than in the controls. The levels of urea nitrogen and creatinine in serum were decreased in rats given ginsenoside-Rd. Decreased urinary levels of glucose, sodium and potassium reflected a protective action against the renal dysfunction caused by cisplatin. In addition, it was demonstrated that ginsenoside-Rd affected cultured proximal tubule cells exposed to cisplatin. PMID- 10803759 TI - Mechanism of reduced GFR in rabbits with ischemic acute renal failure. AB - A reduction in glomerular filtration rate (GFR) is a primary characteristic of ischemic acute renal failure. The present study was undertaken to examine the roles of angiotensin II, tubuloglomerular-feedback (TGF) mechanism, and tubular obstruction for the GFR reduction in the post-ischemic kidney. Renal ischemia was induced by occlusion of the bilateral renal arteries for 60 min, and renal function was examined at 2 and 24 h after the onset of reflow. After the end of 2 h reflow, the GFR was not significantly changed, but the urine flow increased significantly. On the other hand, at the end of 24-h reflow, the GFR and urine flow decreased markedly along with increased filtration fraction. The renal blood flow significantly decreased at 24 h, but not 2 h, after reflow, which was accompanied by increased total renal vascular resistance. Furosemide infusion (1 mg/min/kg) after 24 h of reflow prevented the reduction in GFR and filtration fraction without no changes in renal blood flow and total renal vascular resistance. Pretreatment of enalapril and losartan did not prevent the reduction in GFR, indicating that angiotensin II was not involved. In morphological examinations, tubular obstruction was seen in the proximal and distal tubules of kidneys both at 2 and 24 h after the onset of reflow. In two rabbits subjected to 48 h of reflow, the tubular obstruction was not observed, despite GFR remained depressed. These results suggest that the late reduction in GFR in postischemic kidneys is not mediated by angiotensin II, but is mediated, at least in part, by the TGF mechanism. The tubular obstruction may be not prerequisite for the GFR reduction in rabbits. PMID- 10803760 TI - Effect of arachidonic acid metabolic inhibitors on hypoxia/reoxygenation-induced renal cell injury. AB - The present study was undertaken to examine the role of arachidonic acid (AA) metabolites in hypoxia/reoxygenation (H/R)-induced renal cell injury in rabbit renal cortical slices using AA metabolic inhibitors. Inhibitors of cyclooxygenase (indomethacin and diclofenac sodium) and lipoxygenase pathways (nordihydroguaiaretic acid, caffeic acid, and eicosapentaenoic acid) reduced H/R induced LDH release in a dose-dependent manner, whereas an inhibitor of cytochrome P-450 monooxygenase pathway ethoxyresorufin was not effective. AA increased LDH release in control slices, and the effect was not altered by indomethacin and nordihydroguaiaretic acid. The protective effect of indomethacin was not affected by addition of PGE2, a main product of cyclooxygenase pathway in the kidney. H2O2-induced LDH release was prevented by inhibitors of lipoxygenase but not by inhibitors of cyclooxygenase and cytochrome P-450 monooxygenase H/R induced LDH release was not altered by iron chelators, phenanthroline and deferoxamine, and a potent antioxidant, N,N'-diphenyl-p-phenylenediamine, suggesting that the H/R-induced cell injury is not attributed to a generation of reactive oxygen species. Morphological studies showed that H/R-induced structural changes including cell necrosis were significantly prevented by indomethacin. These results suggest that inhibitors of cyclooxygenase and lipoxygenase pathways exert a direct protective effect against the H/R-induced cell injury in renal tubules. Whether these effects are mediated by alterations of AA metabolic pathways is not certain. PMID- 10803761 TI - Caffeine augments proteinuria in puromycin-aminonucleoside nephrotic rats. AB - Several studies indicate that increased intrarenal adenosine concentrations may attenuate puromycin-aminonucleoside (PAN)-induced nephropathy in rats. The purpose of this study was to investigate the chronic effects of caffeine, a nonselective adenosine receptor antagonist, on renal function and structure in PAN-induced nephropathy. Animals were randomized to receive drinking water or 0.1% caffeine solution. PAN was administered in two doses to a subset from each group at 1 week (100 mg/kg, s.c.; Purom-1) and 15 wks (80 mg/kg, s.c.; Purom-2) after initiating caffeine treatment (PAN and CAFF-PAN groups). The remaining animals served as time controls (CON and CAFF groups). Renal excretory function was followed for 23 wks. Caffeine consumption significantly augmented PAN-induced proteinuria after both PAN injections (Purom-1 and Purom-2, p<0.05 and p<0.001 respectively; CAFF-PAN vs. PAN). In addition, caffeine potentiated the transient reduction in creatinine clearance (CrCl) induced by PAN. Caffeine consumption for 23 wks significantly reduced CrCl in conscious nephrotic animals (4.76 +/- 0.98 vs. 8.51 +/- 1.55 L/kg/day, CAFF-PAN vs. PAN). Seven days after both PAN injections, increased plasma renin activity was detected in animals that were consuming caffeine as compared with corresponding control groups (CAFF and CAFF + PAN vs CON and PAN, respectively). Eight weeks after the second injection of PAN, acute measures of renal hemodynamic and excretory function were compared in anesthetized animals and renal samples were analyzed for histological changes. In PAN-rats, caffeine treatment for 23 weeks significantly reduced inulin clearance (0.28 +/- 0.09 vs. 0.57 +/- 0.12 mL/min/gr kidney. CAFF-PAN vs PAN, p<0.05), tended to increase renal vascular resistance (59.0 +/- 9.5 vs. 42.9 +/- 5.5 mmHg/mL/min/gr kidney, CAFF-PAN vs. PAN, p < 0.06), potentiated the development of more severe tubulointerstitial damage (tubular atrophy, presence of proteinaceous material, tubular dilatation, interstitial inflammation, interstitial fibrosis), and tended to increase glomerulosclerosis. In conclusion, this study indicates that caffeine adversely affects renal function in PAN nephrotic rats, and that this effect may be due, in part, to increased activity of the renin angiotensin system. PMID- 10803762 TI - Factors that affect absorption behavior of cyclosporin a in gentamicin-induced acute renal failure in rats. AB - Factors that affect the absorption of cyclosporin A (CsA) were examined in gentamicin-induced acute renal failure (ARF) rats. In ARF rats, the area under the blood CsA concentration-time curve after oral administration was significantly decreased in comparison with that of control rats; 5.81 +/- 0.55 vs 11.30 +/- 1.59 mg h mL(-1)(mean+/-s.e.m.), respectively, and the relative bioavailabilities in ARF and control rats after oral administration were 15.2% and 43.4%, respectively. The flow rate of bile and the amount of bile acids in ARF rats were markedly decreased to about 61% of control, and 41% of control, respectively. The amount of CsA uptaken into the evened sac of jejunum, transferred to serosal side, and metabolized in tissues was significantly decreased in ARF rats without verapamil, while with 0.3 mM verapamil, the amount in ARF rats recovered to the levels of control rats. The absorption clearance of CsA in ARF rats was significantly decreased, however it was significantly improved by adding bile or bile acid. Adenosine triphosphate released from enterocytes in ARF rats was significantly decreased in the presence of 2.0 microM CsA, 0.3 mM verapamil, or both, in comparison with control rats. From these findings, we concluded that a reduction of CsA bioavailability during ARF is caused by depression in bile excretion and renal function-dependent depression of uptake from intestinal tract via maybe P-gLycoprotein in enterocytes. They are main two factors that reduce the absorbed fraction of CsA in ARF rats. PMID- 10803763 TI - Relationship between IL-1beta and TNFalpha production by PBMC and clinical features of hemodialysis patients. AB - The relationship between production of IL-1beta and TNFalpha by peripheral blood mononuclear cells (PBMC) of hemodialysis (HD) patients and clinical disorders characteristic for HD patients was examined. The study included 28 HD patients divided in the 4 groups: group 1--6 patients with malnutrition; group 2--6 patients with secondary hyperparathyroidism; group 3--6 patients with eosinophilia; group 4--10 stable HD patients without clinical complication. The control group consisted of 9 healthy volunteers. All patients were dialyzed with cuprophane membrane more than one years. Blood samples were taken immediately before the beginning of hemodialysis and PBMC were isolated by centrifugation on the density gradient. Concentrations of IL-1beta and TNFalpha were measured in the supernatants of the cultures by ELISA tests. The results showed marked individual differences in cytokine production both in healthy controls and in HD patients. Spontaneous and LPS stimulated production of IL-1beta by PBMC of HD patients was significantly higher compared to PBMC of healthy controls. There were no significant differences between group of patients with different clinical complications in cytokine production. However, a positive correlation was found between IL-1beta concentration and body mass index for patients with malnutrition and between concentration of IL-1alpha and parathyroid hormone (PTH) for patients with secondary hyperparathyroidism. The results suggest that PBMC of HD patients are chronically stimulated to produce IL-1beta, which may contribute to the development of particular chronic complications. PMID- 10803764 TI - Normalization of hematocrit in hemodialysis patients does not affect silent ischemia. AB - Transient ST-segment depression measured on ambulatory ECG monitors has been described as representing silent ischemia. Patients who demonstrate silent ischemia have been reported to show increased mortality compared to patients without silent ischemia. We undertook this study to determine if the correction of anemia in End Stage Renal Disease (ESRD) patients from (+/- = standard deviation) 30 +/- 3 to 42 +/- 3 with the use of Epoietin alfa would result in decreased silent ischemia in patients with clinically evident ischemic heart disease or congestive heart failure. METHODS: Thirty one ESRD patients with congestive heart failure or patients with clinically-evident ischemic heart disease were randomized into one of two arms. Patients in Group A had their hematocrit increased with the use of slowly escalating doses of Epoietin alfa to 42 +/- 3% and patients in Group B were maintained with a hematocrit of 30 +/- 3% throughout the course of the study. All patients had a 24 hour Holter monitor recording at baseline and at 28 weeks after randomization (when they had reached their target hematocrit). Significant silent ischemia was considered to be present if patients demonstrated at least 60 seconds of > or = 1 mm ST segment depression. RESULTS: Fifteen patients were randomized to Group A and 16 patients were randomized to Group B. The mean hematocrit increased in group A from 29.1 +/ 2.4% to 40.8 +/- 5.2% after 30 weeks. The mean hematocrit in Group B remained stable at 30 +/- 3% throughout the course of the study. Ten patients demonstrated silent ischemia at baseline. At follow up patients in group A demonstrated a mean of 1.7 +/- 4.9 minutes of ischemia compared to 1.1 +/- 3.4 minutes in group B. These were not significantly different. A similar number of patients in group A and Group B required adjustments in their anti-anginal medication during the course of the study. CONCLUSION: It is possible to increase hematocrit to near normal levels in hemodialysis with the administration of exogenous Epoietin alfa. The increase in hematocrit form 30 +/- 3% to 42 +/- 3% is not associated with a change in the level of silent ischemia these patients demonstrate. PMID- 10803765 TI - Enhanced tumor necrosis factor in the serum and renal hypoperfusion in nephrosis associated with focal segmental glomerulosclerosis. AB - Enhanced tumor necrosis factor alpha associated with immunocirculatory imbalance expressed as increased ratio between proinflammatory (TNFalpha) and antiinflammatory (IL-10) cytokines was observed in the serum of nephrosis associated with focal segmental glomerulosclerosis. Such altered immunocirculatory balance correlated with the reduction in renal plasma flow determined by the intrarenal hemodynamic study by which it implies that a glomerular endothelial cell injury associated with impaired renal perfusion is likely to be spontaneously induced by enhanced tumor necrosis factor in the presence of inadequate release of antiinflammatory cytokine (IL-10). PMID- 10803766 TI - Hypoglycemia in patients with renal failure. AB - Of a total of 1545 admissions of end stage renal failure (ESRD) patients, fifty six (3.6%) were admitted during a ten-year period with hypoglycemia. Thirty-eight of them were diabetic while the others were non-diabetic patients. The most common etiology found to be drug-induced hypoglycemia in 26 (46%) patients. In 22 (39%) cases, sepsis was the contributing cause of hypoglycemia. Severe malnutrition caused 7% of hypoglycemic episodes. Of the patients, 18 (32%) with ESRD eventually died, none of them were from the drug induced group. However, mortality rate in the sepsis induced hypoglycemia group was 66%, and of the malnutrition group 17% of the deaths. Thus, hypoglycemia is frequent in ESRD patients, and is fatal if associated with either sepsis or malnutrition. PMID- 10803767 TI - Polymyxin b-immobilized fiber reduces increased plasma endothelin-1 concentrations in hemodialysis patients with sepsis. AB - We studied whether plasma endothelin (ET)-1 concentrations are altered in patients with septic shock who are undergoing hemodialysis and whether polymyxin B-immobilized fiber (PMX-F) treatment affects on these concentrations. Fifteen hemodialysis patients with septic shock treated with PMX-F (group A), 10 such patients who received conventional treatments (group B), 20 hemodialysis patients without septic shock (group C) and 20 healthy controls (group D) were included in this study. Plasma ET1 levels were measured by radioimmunoassay and endotoxin levels were determined by endospecy test. The survival rate in group A (67%) was higher than that in group B (30%). Blood endotoxin levels decreased significantly from 36.4+/-8.2 pg/mL to 10.6+/-3.8 pg/mL (p < 0.01) after PMX-F treatment in group A. The pretreatment plasma ET-1 levels in patients in group A (58.6+/-9.8 pg/mL) and group B (56.8+/-7.8 pg/mL) were significantly higher than those in group C (p < 0.01) and group D (p < 0.001). Plasma ET-1 levels in group C (11.2+/ 3.2 pg/mL) were higher than those in group D (2.6+/-0.6 pg/mL) (p < 0.01). Plasma ET-1 levels following hemodialysis (10.9+/-3.0 pg/mL) were not altered significantly compared with those before hemodialysis. Plasma ET-1 levels decreased significantly in group A after PMX-F treatment (11.4+/-3.6 pg/mL) (p < 0.01); the levels in group B were not altered after conventional treatment. Our data suggest that ET-1 may be associated with septic shock in patients undergoing hemodialysis and that PMX-F is effective in reducing plasma ET-1 levels in these patients. PMID- 10803768 TI - Prediction of acute renal failure by "bedside formula" in medical and surgical intensive care patients. AB - BACKGROUND: Prediction of which intensive care unit (ICU) patients are likely to develop acute renal failure (ARF) would be useful. However, scoring systems such as APACHE have been disappointing in this regard. We previously developed a bedside formula to predict ARF using only 3 parameters: serum albumin, urine osmolality, and presence of sepsis. METHODS: We prospectively evaluated 115 consecutive medical ICU (MICU) patients, comparing the bedside formula to APACHE II AND APACHE III as predictors of ARF or death and looking at nutritional parameters such as iron binding capacity, triceps skin fold, mid-arm circumference, and total lymphocyte count. We then evaluated 123 additional consecutive MICU and 98 consecutive surgical ICU (SICU) patients, comparing the bedside formula to APACHE II. RESULTS: The bedside formula was consistently more accurate than APACHE II in predicting ARF or in-hospital death in MICU patients. However, in SICU neither formula predicted ARF, and APACHE II predicted in hospital death slightly better. No nutritional parameter other than albumin correlated with ARF. CONCLUSION: The bedside formula appears superior to APACHE II in predicting ARF or death in MICU but not SICU. This suggests that these two ICU populations are different. PMID- 10803769 TI - Cephotaxime-associated allergic interstitial nephritis and MPO-ANCA positive vasculitis. AB - We report a case of reversible acute renal failure after cefotaxime treatment in a patient affected by non-Hodgkin lymphoma. Renal biopsy showed necrotizing vasculitis associated with eosinophil-rich interstitial inflammatory infiltrates and patchy infiltrates of CD20+ lymphoid cells. High serum p-ANCA titers were also detected. Drug withdrawal was closely related with recovery of renal function and disappearance of ANCA. Acute renal failure therefore represented a consequence of ANCA-mediated renal vasculitis and acute interstitial nephritis related to cefotaxime treatment. PMID- 10803770 TI - Genetic polymorphism of the CCK gene in patients with alcohol withdrawal symptoms. AB - BACKGROUND: Cholecystokinin (CCK) is considered to play an important role in the central nervous system via its interaction with other neurotransmitters such as dopamine, serotonin, gamma-aminobutyric acid, substance P, and enkephalins. We investigated the relationship between the C to T substitution in the Sp1 binding cis-element of the CCK gene promoter region (at position -45 numbered from initiation codon) and alcohol withdrawal symptoms. METHODS: We examined 214 Japanese men with alcoholism (93 with delirium tremens, 49 with hallucination, 38 with seizure, and 93 with none of these symptoms) and 98 age-matched Japanese male controls by using a polymerase chain reaction-based single strand conformational polymorphism analysis. RESULTS: Patients who displayed hallucination were significantly more likely to possess the C allele than control subjects (chi2 = 8.17, p = 0.017, Bonferroni correction: p = 0.064). In addition, we investigated the influence of CCK gene polymorphism on alcohol consumption among the control subjects but found no significant relationship. CONCLUSIONS: Our data suggested that the C allele at -45 locus of the CCK gene was higher in patients with hallucination than the control group at a rate that was not quite significant after Bonferroni correction for multiple testing. PMID- 10803771 TI - Inactive aldehyde dehydrogenase 2 worsens glycemic control in patients with type 2 diabetes mellitus who drink low to moderate amounts of alcohol. AB - BACKGROUND: Alcohol intake can have hypoglycemic or hyperglycemic effects in patients with type 2 diabetes mellitus. The present study was designed to investigate the glycemic control of male patients with diabetes mellitus from the aspect of the genetic status of alcohol metabolism. METHODS: One hundred sixty three men with type 2 diabetes mellitus were enrolled in the present study. They were all outpatients at the Diabetes Center of Saiseikai Central Hospital. The genotype of the aldehyde dehydrogenase 2 (ALDH2) gene of each patient was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and the patients were divided into those with active or inactive ALDH2 phenotype. We compared the amount of habitual alcohol intake and clinical data that included physical findings and blood chemistry of the patients in the active and inactive ALDH2 groups. The glycemic control of each patient was evaluated by the serum level of HbAlc. RESULTS: Of the 163 patients with type 2 diabetes mellitus, 90 patients had the active ALDH2 phenotype and 73 patients had the inactive ALDH2 phenotype. The mean HbA1c level of the active ALDH2 group was nearly the same as that of the inactive ALDH2 group. However, the HbA1c level of the light-to-moderate drinkers (1-400 g/week) in the inactive ALDH2 group was highest and was significantly higher than the HbA1c level of the light-to moderate drinkers of the active ALDH2 group. The HbA1c of the patients with diabetic complications was higher than the HbAlc of those without diabetic complications in both the active and inactive ALDH2 groups. However, the HbA1c level of the light-to-moderate drinkers without diabetic complications in the inactive ALDH2 group was significantly higher and the incidence of 24 hr urinary C-peptide was higher than the respective level of the light-to-moderate drinkers without diabetic complications in the active ALDH2 group. CONCLUSIONS: Habitual light-to-moderate alcohol intake worsens glycemic control in diabetic patients who have the inactive ALDH2 phenotype. The data on 24 hr urinary C-peptide level suggested that increased acetaldehyde after light-to-moderate drinking by inactive ALDH2 diabetic patients may increase the HbA1c value by the insulin resistant condition that resulted in hyperinsulinemia. PMID- 10803773 TI - Michaelis-Menten elimination kinetics of acetate during ethanol oxidation. AB - BACKGROUND: The redistribution of acetate cannot be explained using a linear kinetic model. We studied the pharmacokinetics of acetate during ethanol oxidation in the rabbit. METHODS: An ethanol saline solution (0.5, 1.5, and 2.5 g/kg) was injected bolus intravenously. We measured blood ethanol, acetaldehyde, and acetate concentrations by using head-space gas chromatography. RESULTS: Blood acetate concentration changed in three phases: an ascending, a plateau, and a declining phase. The first-order rate constant of the declining phase was smaller than that of the ascending phase and decreased dose dependently. Statistical moment analysis of the blood acetate profiles showed that the normalized area under the curve (AUC/Dose) and the mean residence time (MRT) increased with increasing dose amount. These increases suggest a capacity-limited elimination of acetate. We attempted simultaneous multiline fitting, using the three blood acetate disappearance curves, to determine the pharmacokinetic model. Consequently, the blood acetate profile was best described by a Michaelis-Menten elimination kinetic model. The Vmax and Km values of acetate elimination were 40.80 +/- 14.10 mM/hr and 0.47 +/- 0.19 mM, respectively. The fraction of a dose of ethanol converted to acetate (f(AcA)) was calculated to be 0.54. The estimated values are average parameter values of three different doses. Fitted curves suggest smaller f(AcA) at a low dose and larger f(AcA) at a higher dose, which indicate increases of accumulation and redistribution of acetate at higher doses. CONCLUSIONS: Acetate elimination during ethanol oxidation obeys capacity-limited kinetics. PMID- 10803772 TI - Study of mitochondrial DNA deletion in alcoholics. AB - BACKGROUND: Recently, it has been reported that single or multiple mitochondrial DNA (Mt-DNA) deletions have been observed frequently in liver tissue and white blood cells (WBC) obtained from patients with alcoholic liver disease (ALD). In this study, we investigated the deletion of the Mt-DNA encoding adenosine triphosphatase (ATPase) region in WBC to clarify whether Mt-DNA heteroplasmy caused by alcohol drinking is reversible. METHODS: Blood samples were obtained from 4 healthy volunteers, 56 patients with ALD, and 106 nonalcoholic healthy controls. The Mt-DNA encoded ATPase region was amplified by polymerase chain reaction (PCR) by using two primers: forward primer, 5'-AACCAACACCTCTTACAGTGA; and reverse primer; 5'-TTGGTGGGTCATTATGTGTTGT. RESULTS: Heteroplasmy was observed in one volunteer on day 3 and in the remaining persons on day 4 after the start of alcohol consumption. Heteroplasmy was observed for another 6 days after alcohol consumption stopped, but on the 7th day it had disappeared in all volunteers. In WBC Mt-DNA obtained from ALD patients within 3 days of abstinence, heteroplasmy was observed in 38 of the 56 patients (67.9%), whereas heteroplasmy was not detected in any healthy subjects. In 10 of the 18 ALD patients (56%) who had heteroplasmy within 3 days of abstinence, heteroplasmy disappeared after 4 weeks of abstinence. CONCLUSION: An acquired mutation of Mt-DNA, at least in the encoding ATPase region, may result from alcohol drinking and may be reversed by stopping drinking. PMID- 10803774 TI - Identification of 7-hydroperoxycholesterol in human liver by liquid chromatography-mass spectrometry. AB - BACKGROUND: A liquid chromatography-mass spectrometry with an atmospheric chemical ionization interface has been developed to identify simultaneously various oxygenated cholesterols in lipid extracts from alcoholic fatty liver. METHODS: We collected human liver tissues from alcoholic and control subjects at autopsy. We purified the lipid extracts with solid phase extraction and injected them onto a spherisorb ODS column, by using methanol that contained 10 mM ammonium acetate as a mobile phase and a flow rate of 0.7 ml/min. RESULTS: We found the presence of 7-hydroperoxycholesterol and other oxysterols (5-cholesten 3beta-ol-7-one, cholest-5-ene-3beta, 7-diol, and 5, 6beta-epoxy-5beta-cholestan 3beta-ol) in alcoholic fatty liver tissues. CONCLUSIONS: We assume that long-term ethanol ingestion leads to excessive oxidative stress and to the accumulation of cholesterol-derived hydroperoxides and oxysterols in alcoholic fatty liver. PMID- 10803775 TI - Simultaneous quantification of various retinoids by high performance liquid chromatography: its relevance to alcohol research. AB - BACKGROUND: We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. METHODS: We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. RESULTS: This condition allowed separation of 13-cis retinoic acid, 9-cis-retinoic acid, all-trans-retinoic acid, 13-cis-retinol, all trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to day reproducibility. Using this system, we examined effects of light exposure on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40% and 60%, respectively. CONCLUSION: The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed. PMID- 10803776 TI - Biological markers of alcoholism with respect to genotypes of low-Km aldehyde dehydrogenase (ALDH2) in Japanese subjects. AB - BACKGROUND: Although the mutant low-Km acetaldehyde dehydrogenase (ALDH2) allele (ALDH2(2)) with reduced capacity to metabolize acetaldehyde offers biological protection against alcoholism and subsequent alcohol-induced organ damage in many individuals, a significant proportion of individuals with heterozygote of the normal and mutant ALDH2 gene (ALDH2(1)/2(2)) consume excessive amounts of alcohol. Indeed, it has been postulated that habitual drinkers with ALDH2(1)/2(2) may be at a higher risk for alcoholic liver disease than those with ALDH2(1)/2(1). In this study, we determined how representative biological markers of alcoholism (gamma-glutamyltransferase [GGT], carbohydrate-deficient transferrin [CDT], and the mean corpuscular volume of erythrocytes [MCV]) differ with respect to the ALDH2 genotypes in Japanese habitual drinkers. METHODS: We obtained genomic DNA samples from 227 Japanese men with various drinking habits. ALDH2 genotypes were determined by allele-specific polymerase chain reaction. GGT, CDT, and MCV were determined and compared between ALDH2(1)/2(1) and ALDH2(1)/2(2) habitual drinkers who consumed more than 66 g of alcohol per day for more than 5 years. We measured CDT by anion-exchange chromatography followed by turbidity immunoassay by using a commercially available assay kit (Axis %CDT TIA). RESULTS: CDT levels were comparable between the two groups. GGT activities were significantly greater in ALDH2(1)/2(1) than in ALDH2(1)/2(2) habitual drinkers (81 +/- 85 vs. 53 +/- 40 IU/liters, p < 0.02). MCV values, on the other hand, were significantly larger in ALDH2(1)/2(2) than in ALDH2(1)/2(1) subjects (98.2 +/- 5.8 vs. 95.8 +/- 4.2 fl, p = 0.02). When we used elevation of either CDT or GGT to detect habitual drinking in ALDH2(1)/2(1) and 2(1)/2(2) subjects, the sensitivities were 57% and 46%, respectively. CDT levels were similar between habitual drinkers with normal aspartate aminotransferase levels and those with elevated levels. CONCLUSION: GGT and MCV, but not CDT, differ with respect to the ALDH2 genotypes in Japanese male habitual drinkers. ALDH2 genotypes should be considered when interpreting data on biological markers of alcoholism. PMID- 10803777 TI - Immunohistochemical study of hyaluronate receptor (CD44) in alcoholic liver disease. AB - BACKGROUND: It has been suggested that the elevation of serum hyaluronate (HA) levels in liver diseases may be due to increased synthesis of HA by hepatic stellate cells or decreased degradation by sinusoidal endothelial cells. The increase in serum HA levels in patients with cirrhosis is thought to be a response to a reduction in HA receptors (CD44) in hepatic sinusoidal endothelial cells. To learn more about how alcohol affects the number and distribution of HA receptors of hepatic sinusoidal endothelial cells, we immunohistochemically studied CD44 levels in liver biopsy obtained from patients with alcoholic liver disease (ALD patients) and also from patients with nonalcoholic liver disease (non-ALD); ALD patients were evaluated when they were currently drinking and again after they became abstinent. Normal liver tissue obtained from three autopsy cases served as a control. METHODS: Liver biopsy specimens were obtained from 18 ALD patients and 12 non-ALD patients. In ALD patients, liver biopsy was performed twice within 3 days and 4 to 8 weeks after abstinence when serum levels of aspartate aminotransferase and alanine aminotransferase became normal. CD44 in liver specimens was stained with anti-CD44 antibody by streptavidine-biotin peroxidase complex. The intensity of the staining of CD44 in liver tissue was determined by a computer-assisted imaging analyzer. We also measured serum levels of CD44 in both ALD and non-ALD patients. RESULTS: The intensity and the number of CD44 staining increased in both ALD and non-ALD patients compared with those in normal liver, which was negative. The staining intensity of CD44 in liver specimens obtained from patients with ALD who were active in alcohol consumption were significantly higher when compared with patients with ALD after abstinence. Serum levels of CD44 in patients with liver disease increased compared with those of healthy subjects. CONCLUSIONS: The results suggest that HA receptors may increase to degrade the increased HA in serum and/or liver. PMID- 10803778 TI - Ethanol patch test: a simple method for identifying the effectiveness of cyanamide in alcoholics. AB - BACKGROUND: To identify the pharmacological effectiveness of cyanamide, 144 alcoholics treated with cyanamide were subjected to a test that used an acetaldehyde dehydrogenase (ALDH) inhibitor, the ethanol patch test, which is considered to be a good indicator of ALDH2 phenotype. METHODS: We placed 100 microl of 70% ethanol on a lint pad and, as a control, placed the same volume of distilled water on a second pad. The ethanol patch test was performed on 144 alcoholics more than 2 weeks after abstinence from alcohol before and after treatment with cyanamide for 1 week. The dose of cyanamide was increased up to 150 mg until the patch test yielded a positive result. RESULTS: In the ethanol patch test, 36 alcoholics (25.0%) gave a positive result before treatment with cyanamide and might have been ALDH2(1)/2(2) heterozygotes. Among 108 alcoholics who were not positive, the distribution of the cyanamide dose that yielded a positive ethanol patch test result was 30 mg in 42 cases (38.9%), 50 mg in 33 cases (30.6%), 70 mg in 5 cases (4.6%), 100 mg in 6 cases (5.6%), and 150 mg in 2 cases (1.9%). Prevalence of liver cirrhosis was significantly higher in alcoholics who showed a positive ethanol patch test result at doses of less than 50 mg cyanamide than those at doses more than 70 mg (p = 0.029). The prevalence of adverse effects was significantly higher in alcoholics who showed a positive ethanol patch test result at doses of more than 70 mg than at doses of less than 50 mg cyanamide (p = 0.002). CONCLUSIONS: The ethanol patch test is a useful method for identifying pharmacological effectiveness of cyanamide and may reduce the prevalence of side effects in cyanamide-treated alcoholics. PMID- 10803779 TI - A study of colloidal scintigraphy in alcoholic liver diseases: discordance from asialo-scintigraphic findings. AB - BACKGROUND: Our study was undertaken to check the discordance between findings from 99mTc-Sn colloidal reticuloendothelial scintigraphy (RESS) and 99mTc-GSA asialo-scintigraphy (GSA, a technique for evaluation of liver parenchymal cell density and function) and to analyze the discordance in relation to functional disturbances. We compared data between patients with alcoholic liver diseases (ALD) and patients with viral liver diseases (VLD). METHODS: The subjects of this study were 40 patients with chronic liver disease (17 with ALD and 23 with VLD). We used the liver uptake ratio of the tracer of the Sn colloid (SnL15, %), the liver uptake rate (GSAL15, %), and the Rmax (an indicator of total liver receptors) as indices of liver scans. RESULTS: GSAL15 was sometimes nondiscordant from SnL15. The patients were divided into two groups: the nondiscordant group (26 cases where the balance/sum of the two variables was <25%) and the discordant group (14 cases where the balance/sum was > or =25%). SnL15 was 41.6 +/- 16.2% in the nondiscordant group and 42.7 +/- 16.3% in the discordant group (p = 0.80). GSAL15 was 34.3 +/- 12.1% in the nondiscordant group and 21.5 +/- 8.1% in the discordant group (p = 0.001). Rmax was 0.33 +/- 0.17 in the nondiscordant group and 0.113 +/- 0.008 in the discordant group (p = 0.002). Thus, the SnL15, as determined by RESS, did not differ significantly between the nondiscordant and discordant groups, whereas GSAL15 was significantly unfavorable in the discordant group as compared with the nondiscordant group. SnL15 as determined by RESS did not differ significantly between the ALD group (40.4 +/- 18.7%) and the VLD group (43.5 +/- 15.0%) (p = 0.59), whereas Rmax as determined by GSA was significantly improved in the ALD group (0.34 +/- 0.20) compared with the VLD group (0.20 +/- 0.4) (p = 0.02). CONCLUSIONS: Liver cell function was lower in cases that showed discordance between liver cell function and reticuloendothelial function compared with cases without such discordance, although reticuloendothelial function did not differ between discordant and nondiscordant groups. Liver cell function was better in cases of ALD than in cases of VLD, whereas reticuloendothelial function did not differ between the ALD group and the VLD group. PMID- 10803780 TI - Plasma endotoxin and serum cytokine levels in patients with alcoholic hepatitis: relation to severity of liver disturbance. AB - BACKGROUND: Endotoxin plays an important role in the initiation and aggravation of alcoholic liver disease. In this study, we evaluated plasma endotoxin levels and serum concentrations of cytokines and lipopolysaccharide binding protein (LBP) during the acute and recovery phase of patients with alcoholic hepatitis; we also explored the prognostic factors associated with a fatal outcome. METHODS: Fourteen patients, consisting of eight patients with alcoholic hepatitis (AH), five cirrhotics with superimposed AH (LC+AH), and one patient with severe alcoholic hepatitis (SAH), were studied. Among these, two with LC+AH died of hepatic failure. RESULTS: Plasma endotoxin levels in the acute phase were higher in patients with AH (184.4 +/- 159.4 pg/ml) and LC+AH (206.9 +/- 174.9 pg/ml) than in healthy subjects (10.4 +/- 5.5 pg/ml, p < 0.001). In particular, in one patient with SAH and one of two nonsurvivors, plasma endotoxin levels were markedly high relative to the other cases. In most survivors, plasma endotoxin levels decreased in the recovery phase, whereas they further increased at the terminal stage in one of two nonsurvivors. Serum interleukin (IL)-6 and IL-8 levels in the acute phase were significantly higher in patients with AH and LC+AH as compared with healthy subjects. These levels were especially high in nonsurvivors and in one patient with SAH. IL-10 increased in two nonsurvivors, one patient with SAH, and one with LC+AH. In the recovery phase, these cytokine levels in survivors tended to decrease, but in nonsurvivors, IL-6 remained high, and IL-8 and IL-10 further increased. Tumor necrosis factor-alpha levels were below the detection limit throughout the course in all patients. Serum lipopolysaccharide binding protein (LBP) generally was elevated in the acute phase and decreased in the recovery phase in all survivors, but in one of the nonsurvivors, LBP was elevated markedly at the terminal stage. In the acute phase, plasma endotoxin levels were correlated positively with white blood cell counts, neutrophil counts, and serum IL-8. IL-8 was correlated positively with neutrophil counts and negatively with serum cholinesterase, hepaplastin test, and serum albumin levels. IL-6 was correlated positively with white blood cell and neutrophil counts, C-reactive protein, and serum total bilirubin and negatively with hepaplastin test and serum total protein levels. Serum LBP was correlated positively with white blood cell and neutrophil counts. CONCLUSIONS: Endotoxemia and related elevation of IL-8 may play an important role in the activation and migration of neutrophils in patients with alcoholic hepatitis. Marked elevation of inflammatory cytokines, IL-6 and IL-8, are related to severity and poor prognosis of alcoholic hepatitis. Serum LBP may serve as an index of inflammatory reaction in alcoholics. PMID- 10803781 TI - Alcohol enhances lipopolysaccharide-induced increases in nitric oxide production by Kupffer cells via mechanisms dependent on endotoxin. AB - BACKGROUND: Ethanol causes both tolerance and sensitization of Kupffer cells. Accordingly, this study examines the effect of acute ethanol consumption on nitric oxide (NO) production from Kupffer cells with or without lipopolysaccharide (LPS) treatment. METHODS: Rats were given ethanol (4 g/kg body weight) intragastrically, and Kupffer cells were isolated 2 and 24 hr later. Some rats were treated for 4 days with 150 mg/kg/day of polymyxin B and 450 mg/kg/day of neomycin to prevent growth of intestinal bacteria, the primary source of endotoxin in the gastrointestinal tract. After addition of LPS, NO was measured by the Griess reaction. RESULTS: Two hours after ethanol administration, LPS induced NO production by Kupffer cells was diminished by 50% but was enhanced 2 fold at 24 hr. Sterilization of the gut with antibiotics blocked this enhancement. CONCLUSIONS: Kupffer cells isolated from rats early after ethanol exhibited tolerance to LPS, whereas sensitization was observed later. It is likely that sensitization to Kupffer cell is caused by gut-derived endotoxin. PMID- 10803782 TI - Glutathione depletion enhances the formation of superoxide anion released into hepatic sinusoids after lipopolysaccharide challenge. AB - BACKGROUND: We examined the effects of glutathione depletion on the level of superoxide anion released into hepatic sinusoids after lipopolysaccharide challenge. METHODS: Rats were given 1 mg/kg of maleic acid diethyl ester to deplete glutathione in vivo and then 0.5 mg/kg body weight of lipopolysaccharide. RESULTS: This treatment significantly depleted serum reduced glutathione (32.7 +/ 1.7 vs. 23.0 +/- 3.2 mM, p = 0.002). However, it did not affect the serum oxidized glutathione concentration (2.88 +/- 0.56 vs. 3.10 +/- 0.78 mM, not significant). The lipopolysaccharide challenge caused significant superoxide anion formation as compared with controls (0.12 +/- 0.04 vs. 0.22 +/- 0.05 o.d., p < 0.001), and it was enhanced significantly by glutathione depletion (0.28 +/- 0.04 o.d., p < 0.05). There were no significant differences in levels of lipopolysaccharide (2142 +/- 452 vs. 2503 +/- 612 pg/ml) and tumor necrosis factor alpha (277 +/- 186 vs. 252 +/- 88 pg/ml) after the lipopolysaccharide challenge between the glutathione-depleted and nondepleted rats. Moreover, the purine nucleoside phosphorylase/glutamic-pyruvic transaminase ratio in liver perfusates, a marker of damage to endothelial cells in hepatic sinusoids, was significantly higher in the glutathione-depleted rats than in the nondepleted rats. CONCLUSIONS: The reduced form of glutathione can decrease levels of the superoxide anion released into hepatic sinusoids and can decrease subsequent damage to endothelial cells in these sinusoids caused by lipopolysaccharide; that is, it can reduce lipopolysaccharide-induced liver injury. PMID- 10803783 TI - Clinical characteristics of patients in their 40s with HCV antibody-positive hepatocellular carcinoma. AB - BACKGROUND: We investigated the clinical characteristics of hepatitis C virus (HCV) antibody-positive hepatocellular carcinoma (HCC) patients who developed HCC at a relatively young age. METHODS: Clinical characteristics of patients in their 40s were investigated and were compared with those of patients 50 years and older. The subjects were 648 HCC patients, 469 men (72%) and 179 women (28%), who were treated at our hospital between 1991 and 1997. RESULTS: No patient was under 40 years of age. Eighteen patients (3%) were in their 40s, 137 patients (21%) were in their 50s, 338 patients (52%) were in their 60s, 143 patients (22%) were in their 70s, and 12 patients (2%) were in their 80s. Fifteen of the patients (83%) in their 40s were male. The proportion of men in their 40s was higher than that of all men. Eight of the 15 men in their 40s (53%) were heavy drinkers, and 2 (14%) were habitual drinkers. Three of the 15 men (20%) were HBV carriers, and these 3 HBV carriers were not drinkers. The proportion of heavy drinkers and HBV carriers was significantly higher among the patients in their 40s than in the 60 patients randomly sampled from the patients 50 years of age and older. The mean ages of male patients with HCC who were heavy drinkers, habitual drinkers, occasional drinkers, or nondrinkers were 52.3, 58.9, 62.0, and 61.7 years, respectively. HCC occurred significantly earlier in heavy drinkers than in the other 3 groups. We compared laboratory data of the patients in their 40s with data of all of the patients of 50 years and older. Serum total bilirubin, prothrombin time, and platelet counts were significantly worse in the patients in their 40s. CONCLUSIONS: Logistic regression analysis revealed that heavy drinking and presence of HBV infection were independently related to HCV antibody-positive HCC development at a younger age. PMID- 10803784 TI - Infiltrating polymorphonuclear leukocytes and apoptotic bodies derived from hepatocytes but not from ballooning hepatocytes containing Mallory bodies show nuclear DNA fragmentation in alcoholic hepatitis. AB - BACKGROUND: The mechanism of liver cell death in alcoholic hepatitis is still controversial. There is evidence that polymorphonuclear leukocytes (PMNs) which infiltrate liver parenchyma are involved in liver cell death. METHODS: We employed terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate digoxigenin nick end labeling (TUNEL) method to determine the cells that underwent apoptosis in liver specimens obtained from 1 normal liver, 23 livers with nonalcoholic liver disease, and 11 livers with alcoholic liver diseases (4 alcoholic hepatic fibrosis, 4 alcoholic hepatitis, and 3 alcoholic cirrhosis). The Fas, bcl-2, and CD15 antigens were immunolocalized to analyze the relationship between these antigens and apoptosis. RESULTS: Few TUNEL-positive reactions were found in normal liver. Apoptotic bodies in acute and chronic viral hepatitis were found to be TUNEL-positive. TUNEL-positive materials were also abundant in the cytoplasm of macrophages at the area of parenchymal collapse in viral liver diseases. Mononuclear cells that infiltrated liver parenchyma and portal tracts occasionally showed TUNEL-positive reactions, which reflected the activation-induced apoptosis of T lymphocytes. In alcoholic liver diseases, the incidence of apoptotic bodies derived from hepatocytes varied from 0 to 1.00/mm2. In addition, a considerable portion (0.2-16.5%) of infiltrating PMNs in alcoholic hepatitis were TUNEL-positive. In contrast, the nuclei of ballooning hepatocytes that contained Mallory bodies revealed few positive TUNEL reactions. Interestingly, the density of CD15-positive PMNs was well correlated with peripheral white blood cell counts (r = 0.842, p < 0.005). CONCLUSIONS: PMNs that infiltrate liver parenchyma show an activation-induced cell death-like phenomenon in alcoholic hepatitis, like activated T cells that infiltrate in viral hepatitis. This apoptosis phenomenon may follow the degranulation of PMNs, which may cause a necrosis of ballooning hepatocytes that contain Mallory bodies through satellitosis. PMID- 10803785 TI - Hepatic veno-occlusive lesions and other histopathological changes of the liver in severe alcoholic hepatitis--a comparative clinicohistopathological study of autopsy cases. AB - BACKGROUND: The clinicopathological features of veno-occlusive lesions (VOL) in the liver were studied in 18 autopsy cases of severe alcoholic hepatitis (sALH). All the cases were heavy drinkers and died of liver failure or variceal rupture. METHODS: We performed histological evaluation by examining stained sections of liver blocks from each case. The severity of VOL was compared with the clinical findings and histopathological changes of alcoholic liver diseases (ALD). RESULTS: Clinically, as the severity of VOL increased, the amount of ascites observed during autopsy increased significantly (p = 0.001) and the time from hospitalization to death was significantly longer (p < 0.05). The peripheral leukocyte count tended to increase and the serum bilirubin level increased significantly (p < 0.05) with increased VOL severity, after we excluded one case that was complicated by acute respiratory distress syndrome and bacterial endocarditis. Histopathologically, the appearance of Mallory bodies increased significantly as VOL became more severe (p < 0.05), but the VOL severity did not correlate with sinusoidal neutrophil infiltration. Fatty degeneration tended to be milder as VOL increased in severity although the difference was not significant, whereas bile retention tended to be more marked. CONCLUSIONS: We conclude that investigation of VOL is clinicopathologically important when assessing the pathophysiology and severity of sALH. PMID- 10803786 TI - Scanning electron microscopic studies on morphological abnormalities of erythrocytes in alcoholic liver diseases. AB - BACKGROUND: We previously reported that morphological abnormalities and the altered deformability of erythrocytes play important roles in disturbances of hepatic microcirculation associated with alcoholic liver disease. METHODS: The subjects of this study were five patients with alcoholic liver cirrhosis, two patients with alcoholic fatty liver, and two healthy volunteers. Concentrating on erythrocyte morphology in the presence of alcoholic liver disease, we observed erythrocytes under a scanning electron microscope and examined their morphological changes in relation to the disease. RESULTS: Among the five patients with alcoholic liver cirrhosis, stomatocytes accounted for about 14% and acanthocytes for about 10% of all erythrocytes. In the fatty liver group (n = 2), acanthocytes were absent, and stomatocytes accounted for 23% in one subject and 56% in the other, of the total. When the two patients with liver cirrhosis were examined over time, one of them was found to have 12% stomatocytes, 21% acanthocytes, and a filtration time of 14.7 sec at admission. Following 1 month of abstinence, this patient had 1% stomatocytes, 6% acanthocytes, and a filtration time of 10.4 sec, which were accompanied by improvements in peripheral blood parameters and liver function. In another patient with liver cirrhosis, there were no acanthocytes, 8% stomatocytes, and a filtration time of 5 sec at admission; this patient had 1% stomatocytes and a filtration time of 7.7 sec after 1 month of abstinence. In one patient with fatty liver who was examined over time, the percentage of stomatocytes was 56%, and the filtration time was 7.7 sec at admission; these parameters were 1% and 5.1 sec, respectively, after 1 month of abstinence. CONCLUSIONS: The present study revealed that stomatocytes and acanthocytes are morphologically abnormal erythrocytes observed in the presence of alcoholic liver disease. These abnormal forms of erythrocytes tended to normalize as peripheral blood parameters and liver function were improved by abstinence, which suggests that erythrocyte morphology is related to the pathophysiology of alcoholic liver disease. PMID- 10803787 TI - Rifampicin suppresses hepatic CYP2E1 expression and minimizes DNA injury caused by carbon tetrachloride in perivenular hepatocytes of mice. AB - BACKGROUND: Rifampicin has been shown to increase during activities of serum transaminases and to decrease in cytochrome P-450-mediated monooxygenase activities in livers of mice treated with carbon tetrachloride (CCl4). Although these findings suggest that rifampicin prevents hepatocyte damage caused by CCl4, detailed information on the protective effects is not available. METHODS: We injected first rifampicin and then CCl4 into mice and examined denaturation and fragmentation of hepatocyte DNA by in situ nick translation, in situ end labeling, and in situ hybridization. Furthermore, expressions of p53, a cytoplasmic marker for apoptosis, and bcl2, an anticell death factor, were examined immunohistochemically. In addition, a major ethanol-inducible P-450 isoform in liver homogenates or microsomes, CYP2E1, was examined by Western blotting, because the enzyme metabolizes CCl4 and forms free radicals to injure perivenular hepatocytes in which the enzyme is restrictedly expressed. RESULTS: Rifampicin prevented the denaturation and fragmentation of DNA caused by CCl4 in perivenular hepatocytes except for those located within two or three cell layers surrounding the central venule. Furthermore, CYP2E1 decreased in liver homogenates or microsomes from rifampicin-treated animals. It is therefore likely that rifampicin suppresses expression of CYP2E1 and protects CCl4-mediated DNA damage of hepatocytes by inhibiting formation of free radicals. In addition, perivenular hepatocytes except for those surrounding the venule showed negative immunoreaction for p53 and bcl2 in rifampicin+CCl4-treated animals. CONCLUSIONS: The drug did not alter the mechanism of cell death from necrosis to apoptosis and did not promote recovery of hepatocytes from CCl4-mediated damage. PMID- 10803788 TI - Immunohistochemical study on acetaldehyde adducts in alcohol-fed mice. AB - BACKGROUND: Acetaldehyde binds to some proteins, which results in Schiff base formation. It is assumed that acetaldehyde binds to the proteins after the consumption of ethanol, to form an adduct. Such acetaldehyde adducts are related to organ disease. METHODS: We examined 8-week-old male BALB/c mice, which were given a liquid diet for 7 days. The diet consisted of vitamins, minerals, amino acids, and a 5% (v/v) ethanol solution. After the 7 days, we took tissue samples from the brain, liver, and adrenal cortex to investigate the distribution of acetaldehyde adducts. We performed immunohistochemical staining of the cerebral cortex, liver, and adrenal cortex from the mice by using antibodies against acetaldehyde adducts. RESULTS: Our study showed that acetaldehyde adducts formed in the cerebral cortex in the early phase in alcohol-fed mice. CONCLUSIONS: Because acetaldehyde in the liver has been shown to cause liver damage, our study suggests a relationship between acetaldehyde adducts in the brain and brain damage. PMID- 10803789 TI - Comparison of cyanamide and disulfiram in effects on liver function. AB - BACKGROUND: Cyanamide, an aversive drug widely used in Japan, develops ground glass inclusion bodies in the hepatocytes at high incidences, which may be associated with portal inflammation and fibrosis. When cyanamide-treated alcoholics relapse drinking, the combined effect of cyanamide and alcohol produce more severe portal inflammation along with the emergence of ground-glass inclusions. Disulfiram also causes hepatitis, but there have been no comparative studies of effects of cyanamide and disulfiram on liver function. METHODS: We reviewed the laboratory data of 408 alcoholics admitted for a 3 month course of alcohol detoxification and rehabilitation. Patients tested negative for hepatitis virus markers and were diagnosed as not having cirrhosis. Among the subjects, 222 patients received cyanamide treatment (a daily dose of 70 mg) without a history of disulfiram treatment, and 186 received disulfiram (a daily dose of 200 mg) without a history of cyanamide treatment. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels obtained at 0, 4, 8, and 12 weeks of administration of each aversive drug were compared between the two alcoholic groups. RESULTS: Elevation of serum transaminases (AST > ALT) probably due to alcoholic liver disease quickly fell after abstinence. In patients who took cyanamide, the ALT levels were significantly higher at 4 and 12 weeks than in those who took disulfiram. Re-elevations of ALT after alcohol detoxification were more frequently observed in those who took cyanamide than in those who took disulfiram (19.4% vs. 5.9%, p < 0.001). The re-elevations of ALT were slight to moderate, being more than 3-fold in three (1.4%) patients who took cyanamide and four (2.2%) who took disulfiram. The re-elevations occurred more frequently in those with a history of cyanamide treatment before the present treatment than in those who took cyanamide for the first time (31.1% vs. 16.4%, p < 0.05). CONCLUSIONS: Cyanamide, compared with disulfiram, was more frequently associated with elevations of ALT that persisted after abstinence. PMID- 10803790 TI - Cyanamide-induced liver dysfunction after abstinence in alcoholics: a long-term follow-up study on four cases. AB - BACKGROUND: Cyanamide, an aversive agent widely used in Japan, is known to induce various degrees of hepatic lesion with ground-glass inclusion bodies. When cyanamide-treated alcoholics relapse into drinking, more severe inflammation develops in the liver. However, it is controversial whether progressive hepatic lesions develop in complete abstainers as a result of long-term cyanamide treatment. CASE REPORTS: Case 1: A 53-year-old male alcoholic received cyanamide treatment for 4.5 months and completely abstained without cyanamide treatment for 6 years. A liver biopsy shortly after abstinence showed extensive pericellular fibrosis, but a biopsy after 6 years showed very mild fibrosis. Case 2: A 43-year old male alcoholic remained completely abstinent with cyanamide treatment for 5 years and complained of general fatigue. His serum transaminases were slightly elevated and hepatic hyperechogenicity was observed on ultrasonography. Only mild pericellular fibrosis was present in the liver biopsy specimen obtained shortly after abstinence, but after 5 years the second liver biopsy showed that thin septum-like fibrosis that formed portal-to-portal and portal-to-central linkage had developed and ground-glass hepatocytes had emerged extensively. Case 3: A 29 year-old female alcoholic complained of general fatigue and a slight fever after 1.5 years of abstinence with cyanamide treatment. Slight elevation of serum transaminases and hepatic hyperechogenicity were observed. The liver biopsy showed extensive ground-glass hepatocytes and thin septum-like fibrosis that formed portal-to-portal linkage. Case 4: A 61-year-old male alcoholic who remained completely abstinent while taking cyanamide for 3 years showed slight elevation of serum transaminases. Liver biopsy showed extensive ground-glass hepatocytes and extension of thin septum-like fibers from portal tract to the lobule. Ultrasonography revealed hepatic hyperechogenicity. CONCLUSION: In some abstainers who take cyanamide for several years, thin septum-like liver fibrosis progresses along with the emergence of ground-glass hepatocytes. Hepatic hyperechogenicity on ultrasonography and slight elevation of serum transaminases might erroneously lead to a diagnosis of hepatic steatosis without liver histology. PMID- 10803791 TI - Effects of alcohol consumption on histological changes in chronic hepatitis C: a clinicopathological study. AB - BACKGROUND: To assess the effects of alcohol on the histological changes in chronic hepatitis type C, we performed histopathological examination on liver biopsy specimens by using a semiquantitative method. METHODS: Subjects were 91 patients with chronic hepatitis type C and 32 with alcoholic liver disease. The patients with chronic hepatitis type C were classified into three groups according to the total amount of alcohol intake: nondrinkers, moderate drinkers, and heavy drinkers. For each patient, we evaluated pathological changes of several items and awarded scores from 0 to 2 points, with severe to moderate scoring 2 points, mild 1, and negative 0; the total score was then compared between groups. The evaluated histological changes included virus-related histological changes (V1, inflammatory cell infiltration; V2, lymphoid follicle formation; and V3, bile duct damage) and alcohol-related changes (A1, perivenular fibrosis; A2, stellate/pericellular fibrosis; and A3, fatty change). RESULTS: The total scores of the hepatitis C virus-related histological changes were significantly lower in patients with alcoholic liver disease (ALD group) (p < 0.05). However, we found no significant difference between the different alcohol intake groups. The total score for alcohol-related histological changes significantly increased in line with increases in total alcoholic intake regardless of the presence or absence of hepatitis type C virus infection (p < 0.05). CONCLUSIONS: The results suggest that both alcoholic-related liver damage and virus-related liver damage have specific features; in a addition, alcohol was found to have little effect on the histological liver damage observed in chronic hepatitis type C. PMID- 10803793 TI - Long-term ethanol consumption in ICR mice causes mammary tumor in females and liver fibrosis in males. AB - BACKGROUND: Although epidemiological studies indicate that ethanol consumption and the risk of breast cancer are positively associated in women, experimental animal models have not yet been developed that provide evidence to support this relationship. To clarify alcohol-related liver injury, it is important to reproduce, in laboratory small animals, the liver fibrosis observed in human alcoholics. However, in mice the induction of fibrosis has failed. The present study describes the first experimental models to produce mammary tumors in female ICR mice and liver fibrosis in male ICR mice treated long-term with ethanol. METHODS: The study consisted of two parts. To induce mammary tumors, female ICR mice were given 10% to 15% ethanol solution as the sole drinking fluid for 25 months, with solid diet supplied ad libitum. To induce liver fibrosis, male ICR mice were given 10% to 15% ethanol solution as the sole drinking fluid for 10 to 15 months. Control female and male mice were given tap water. RESULTS: In 9 (45%) of 20 ethanol-treated female mice, mammary tumors occurred at 8 to 24 months after ethanol intake began, whereas spontaneous mammary tumor was not found in the 20 control female mice. The tumors were composed histopathologically of either papillary adenocarcinoma or medullary adenocarcinoma of glandular epithelial origin. In the ethanol-treated male mice, early hepatic fibrosis at the centrilobular and pericellular areas and central-central bridging were observed at the 10th month, and marked fibrosis at the centrilobular, pericellular, and periportal areas and bridging between the neighboring vascular tissues were observed at the 15th month, which suggested that the initial fibrosis arose from the centrilobular area. No abnormalities other than mild fatty infiltration were found in livers of the control male mice. CONCLUSIONS: These murine models may be useful to study the role of ethanol in mammary tumorigenesis and the pathogenetic mechanisms of ethanol liver injury. PMID- 10803792 TI - Contribution of hepatitis C virus to the progression of alcoholic liver disease. AB - BACKGROUND: We determined hepatitis C virus (HCV) antibody, HCV RNA, and genotype in patients with alcoholic liver disease and studied the involvement of HCV in alcoholic liver disease. Additionally, we used the histological activity index (HAI) to study the influence of HCV on the severity of inflammation. METHODS: The subjects were 143 patients with alcoholic liver disease: 7 with fatty liver (FL), 18 with hepatic fibrosis (HF), 24 with alcoholic hepatitis (ALH), 39 with chronic hepatitis (CH), 42 with liver cirrhosis (LC), and 13 with hepatocellular carcinoma (HCC). The HCV RNA positivity rate in each type of disease was 0/7 (0%), 1/18 (6%), 2/24 (8%), 27/39 (69%), 24/42 (57%), and 7/13 (54%), respectively. It was high in the advanced hepatic lesions. RESULTS: Clinically, the serum hepatic function tests after abstinence from drinking improved significantly in the HCV RNA negative patients compared with the positive patients. The proportion of genotype II in each type of disease was 0/0, 0/1 (0%), 1/2 (50%), 18/27 (67%), 18/24 (75%), and 7/7 (100%), respectively. It became high with the advance of pathophysiology. The HCV RNA amount stood at 7.5 +/- 0.4 [log (copies/ml)] in CH, 7.9 +/- 0.4 in LC, and 8.4 +/- 0.8 in HCC, with a statistically significant difference between CH and HCC. However, we found no changes in the HCV RNA amount due to abstinence from drinking. The HAI score was high in the HCV RNA positive patients, but several cases in the HCV RNA negative group showed severe inflammatory changes. Therefore, judging the presence or absence of HCV RNA with the HAI score alone was considered difficult. CONCLUSIONS: These results suggest that HCV, particularly genotype II, plays an important role in the advance of disease to LC and HCC in heavy drinkers. PMID- 10803794 TI - Women with epilepsy. PMID- 10803795 TI - Cerebral ischemia-reperfusion injury: a novel therapeutic approach with TAK-218. AB - The goal of the present study was to evaluate the potential neuroprotective effect of TAK-218 in an in vivo rat focal cerebral ischemia/reperfusion model. TAK-218 is a novel compound with multiple antiischemic properties, including suppression of aberrant dopamine release, modulation of sodium channels, and inhibition of lipid peroxidation. The study was a blinded, randomized, placebo controlled study of TAK-218 in a three-vessel focal ischemic rat model. A total of 22 rats were randomly assigned to the treatment or placebo group. Animals were injected intrapertoneally with either a 2 mg/kg dose of drug or saline at 2 hours after reperfusion. Infarction volume was measured with use of 2,3,5 triphenyltetrazolium chloride. Total adjusted infarction volume in treated animals decreased by 10%. With use of a statistical analysis requiring 80% power with a 20% reduction desired effect, there was no statistically significant difference in the end-point of infarction volume between drug and placebo treatment groups. In light of the proven efficacy of thrombolytic therapy for acute stroke, it is now desirable to test neuroprotective agents during the 3 hour therapeutic window after ischemia. Further research is necessary to discern if a therapeutic agent with multiple antiischemic properties may provide a more robust neuroprotective effect than an agent with a single neuroprotective action. PMID- 10803796 TI - A home diary to assess functional status in patients with Parkinson's disease with motor fluctuations and dyskinesia. AB - In clinical trials for patients with Parkinson's disease (PD) with motor fluctuations, efficacy is generally ascribed to an intervention if motor function is significantly improved or if "off" time is significantly reduced. However, we have argued that patients might not be improved if off time is reduced only to the extent that unwanted dyskinesia is increased. Therefore, a home diary should include an assessment of dyskinesia to provide an accurate reflection of clinical status over a period of time. We undertook two studies to develop a home diary to assess functional status in patients with PD with motor fluctuations and dyskinesia. In both studies, patients concurrently completed a test and a reference diary. In Study I, we evaluated the impact of different severities of dyskinesia on patient-defined functional status. There were 1,149 evaluable half hour time periods from 24 patients; 94.3% of off time was considered "bad" time and 90.2% of "on" time without dyskinesia, 72.6% of on time with mild dyskinesia, 43.0% of on time with moderate dyskinesia, and 15.2% of on time with severe dyskinesia was considered "good" time. In Study II, we evaluated a new home diary designed to separate dyskinesia that had a negative impact on patient-defined functional status from dyskinesia that did not. There were 816 evaluable time periods from 17 patients; 84.9% of off time and 89.9% of on time with troublesome dyskinesia was considered bad time while 85.5% of on time without dyskinesia and 93.8% of on time with nontroublesome dyskinesia was considered good time. With this diary (Diary II), the effect of an intervention can be expressed as the change in off time and the change in on time with troublesome dyskinesia (bad time). The sum can be used as an outcome variable and compared to baseline or across groups. In evaluating the efficacy of an intervention, assessment of change in off time and change in on time with troublesome dyskinesia provides a more accurate reflection of clinical response than change in off time alone. PMID- 10803797 TI - The effect of amantadine on levodopa-induced dyskinesias in Parkinson's disease: a double-blind, placebo-controlled study. AB - We performed a double-blind, placebo-controlled, crossover study to assess the effect of amantadine versus placebo on levodopa-induced dyskinesias in Parkinson's disease. We found a 24% reduction in the total dyskinesia score after amantadine administration (p = 0.004). This improvement was achieved without any influence on the severity of "on" period parkinsonism. The results confirm that amantadine reduces levodopa dyskinesias and support the hypothesis that dyskinesias can be reduced by blockade of excitatory pathways in the basal ganglia. PMID- 10803798 TI - Disabling erratic myoclonus during lamotrigine therapy with high serum level- report of two cases. AB - We present two patients with epilepsy who experienced disabling myoclonic jerks during lamotrigine treatment. Both were young males who had intractable cryptogenic generalized epilepsy since childhood. They received a lamotrigine valproate combination resulting in an excellent improvement; however, after 2-3 years of therapy, both patients were hospitalized because of continuous disabling myoclonic jerks. The dosage of lamotrigine was the same before and at the onset of myoclonus. When the severe myoclonus started, both patients had a higher serum lamotrigine level (16.5 and 17.7 mg/L, respectively) than in previous findings. Disabling myoclonus was also present during lamotrigine monotherapy with 15 mg/L serum level. Lamotrigine may severely worsen myoclonic phenomena in generalized epilepsies, in which adverse events may be dependent on drug serum level. PMID- 10803799 TI - Neuropharmacology of paradoxic weight gain with selective serotonin reuptake inhibitors. AB - It has been suggested that weight gain associated with tricyclic antidepressants (TCA) reflect actions on dopamine (DA) and histamine receptors. However, a definitive cause is purely assumptive given the nonselective pharmacology of these agents. The selective serotonin reuptake inhibitors (SSRIs), as well as agents like dexfenfluramine (DFF), have emphasized the pivotal role of serotonin (5HT) in reducing carbohydrate (CHO) intake, and have provided a more selective tool with which to study appetite regulation. It would be expected that all SSRIs should exert a similar anorectic action. However, recent reports provide evidence to the contrary. Despite their claimed selectivity, SSRIs still interact, either directly or indirectly, with various critical neurotransmitter systems. In addition, although the anorectic action of fluoxetine (FLX) is well recognized, long-term follow-up studies in depressed patients and in obese nondepressed patients reveal that its weight-reducing effects are transient, even leading to a gain in body weight. Similarly, paroxetine (PRX) and citalopram (CTP) have also been associated with weight gain. These latter observations are unexpected because PRX and CTP are highly potent and selective SSRIs. A neuropharmacologic rationale for the apparent paradoxic effects of SSRIs on appetite not a review of neuronal regulation of appetite is presented in this article. As with the regulation of feeding, paradoxic weight gain observed with SSRIs appears to rest on the interaction of 5HT with multiple mechanisms, with the extent of weight gain observed being dependent on subtle, yet important pharmacologic differences within the group. Finally, the neurobiology of depressive illness itself, and of recovery from it, is a major contributing factor to individual response to these drugs. PMID- 10803800 TI - Clinical, pharmacokinetic, and pharmacodynamic effects of tolcapone withdrawal in levodopa-treated patients with parkinsonism. AB - The effect and clinical significance of tolcapone withdrawal on erythrocyte catechol-O-methyltransferase (COMT) activity, levodopa pharmacokinetics, and levodopa requirements were investigated in 59 patients with fluctuating parkinsonism who were randomized to receive placebo or tolcapone 100 or 200 mg three times daily for 6 weeks. Tolcapone withdrawal caused a transient elevation in COMT activity by 64% in patients receiving 100 mg three times daily and by 128% in those receiving 200 mg three times daily at approximately 1-2 weeks after discontinuation of drug. Thereafter, COMT activity was declining but did not reach baseline values by the 12-week study endpoint. However, this had no effect on plasma levodopa and 3-O-methyldopa (3-OMD) concentrations or on levodopa requirements. During treatment, tolcapone increased "on" time and decreased "off" time; after discontinuation of study medication and levodopa dose adjustment, on and off times were similar to baseline. Withdrawal was generally well tolerated; no patients withdrew from the trial during the posttreatment period, and no serious adverse events were observed. In conclusion, the transient increase in erythrocyte COMT activity observed after discontinuation oftolcapone is not associated with changes in peripheral levodopa metabolism and therefore has no significant clinical consequences in terms of levodopa requirements, clinical symptoms, or adverse events. PMID- 10803801 TI - Regulation of phosphorylation of cyclic AMP response element-binding protein by paroxetine treatments. AB - The present study was undertaken to examine the effect of paroxetine, a selective serotonin reuptake inhibitor, on the phosphorylation of cyclic adenosine monophosphate (cAMP) response element binding protein (CREB) in the rat brain. Single administration of paroxetine significantly induces the phosphorylation of CREB in the rat frontal cortex and hippocampus in a time-dependent manner. Repeated administration of paroxetine attenuates CREB phosphorylation in response to acute paroxetine challenge. These findings suggest that the enhancement of intracellular signal transduction after the activation of serotonin receptors may be attenuated after chronic paroxetine treatment, and this attenuation may be, at least in part, involved in the therapeutic efficacy of paroxetine. PMID- 10803802 TI - Immunoglobulins are effective in pontine myelinolysis. AB - Although the exact pathogenesis of central pontine myelinolysis (CPM) is unknown, correction of hyponatremia, thyreotropin releasing hormone, plasmapheresis, and corticosteroids seem to be effective. Assuming intravenous immunoglobulins (IVIG) to also be effective in CPM, 0.4 g/kg body weight/d immunoglobulins were applied to a 48-year-old patient who developed CPM with double vision, dysarthria, dysphagia, and left-sided hemiparesis 3 weeks after spontaneous normalization of hyponatremia. After 5 days of IVIG, his symptoms markedly improved, confirmed by improvement in the Norris score (42%), Frenchay score (19%), Kurtzke score (20%), Disability score (54%), vital capacity (26%), and peak torque (69%). The promising clinical effect of IVIG was assumed to be caused by the reduction of myelinotoxic substances, the development of antimyelin antibodies, and the promotion of remyelination. In conclusion, IVIG appear to be a promising therapeutic option in CPM. PMID- 10803803 TI - Double-blind, crossover, placebo-controlled clinical trial with L-acetylcarnitine in patients with degenerative cerebellar ataxia. AB - Despite the different genetic defects underlying degenerative ataxias, it has been suggested that mitochondrial energy production and antioxidative metabolism dysfunction may be common biochemical alterations related to these diseases. Acetylcarnitine, a cholinomimetic substance, is involved in oxidative metabolism and is a potential source of acetyl groups for the synthesis of acetylcholine in the mammalian brain. To determine whether treatment with L-acetylcarnitine may improve some clinical conditions of patients with ataxia, a double-blind crossover study with L-acetylcarnitine was performed in 24 patients with degenerative cerebellar diseases. Patients were selected from an ongoing prospective follow-up study at the Department of Neurology at the University of Florence, Italy. Each treatment phase with L-acetylcarnitine or placebo lasted 6 months, after which patients were crossed over to the other treatment phase. Ataxia was documented and quantified with use of a clinical score. After the trial, we observed a statistically significant improvement of some symptoms and a slow progression of the disease in both groups of patients. PMID- 10803804 TI - Parental bonding and personality in relation to a lifetime history of depression. AB - This study explores whether personality is mediating the effects of adverse parenting on having had a lifetime history of major depressive disorder and whether personality dimensions, related to the development of lifetime depression, are disposed by adverse parenting in cross-sectional data derived from an epidemiological sample of volunteer workers. Of 447 individuals who were asked to complete the Munich Personality Test (MPT), the Parental Bonding Instrument (PBI) and the Inventory to Diagnose Depression Lifetime version (IDDL), 322 subjects were included in the analyses (150 male and 172 female; and 38 were diagnosed as having had a history of depression). Comparisons in fit between logistic regression models revealed that a combination of frustration tolerance and rigidity among personality dimensions, as measured by the MPT, and maternal care among the PBI scales were most primary in predicting a lifetime history of depression. Maternal care was, however, not significantly predictive of dimensional scores on the personality dimensions. Neither frustration tolerance nor rigidity was predicted by any PBI scale. When entering the variables sequentially, maternal care and the personality variables were additive and independent risk factors in predicting a lifetime history of depression. The results of this preliminary study raised an objection to a hypothesis that adverse parenting experienced in childhood disposes one to a dysfunctional personality, which then predisposes one to the development of depression in adulthood. PMID- 10803805 TI - Chronological progression of a language deficit appearing to be postictally reversible in a patient with symptomatic localization-related epilepsy. AB - A language deficit occurring interictally, with chronological progression, and postictally in a patient with symptomatic localization-related epilepsy, which began at 1.6 years of age, is reported. The patient was a 30-year-old right handed man whose seizures seemed to originate from the left frontal lobe and to involve the left temporal lobe. The deficit in oral language consisted mainly of features of motor aphasia, including delayed initiation of speech with great effort, echolalic and palilalic tendencies, and word-finding difficulty, but he also showed features of sensory aphasia. Written language had agraphia observed in sensory aphasia, including well-formed letters, paraphasias, neologisms, and paragrammatism. Postictally, the language deficit appeared to be superficially reversible, and evolved from mutism through non-fluent jargon to the interictal level of language. Analysis of the patient's diaries from 10 to 26 years of age disclosed chronologically progressive deterioration of language with paragrammatism, showing an increase of grammatical errors, neologismus, literal and verbal paraphasias and misconstruction of sentences. The results suggest that localization-related epilepsy of presumably left frontal lobe origin causes not only a postictal language deficit but also a slowly progressive deficit of language function. PMID- 10803806 TI - Adolescent hyperactivity and general psychopathology. AB - The aim of this study was to explore the relationship between activity and psychopathology in adolescents. A total of 289 high school students completed the Symptom Check List-90-R (SCL-90-R) and Wender Utah Rating Scale (WURS) in December 1998. The WURS was used to measure the students' activity level and the SCL-90-R was used as a measure of general psychopathology. Forty-one students (14.18%) scored higher than 46, the cut-off point for differentiation of attention deficit hyperactivity disorder (ADHD) from the general population according to Ward's report, in the WURS (WURS(+)). The WURS(+) students scored significantly higher than the WURS(-) students in all the subscales of SCL-90-R. The prevalence rate of adolescent WURS(+) in this study is 14.18%. This result shows that ADHD adolescents have overlapping symptoms with depression. PMID- 10803807 TI - Comparative study of quality of life between the Chinese and Japanese adolescent populations. AB - Researchers from Japan, China and Singapore have initiated a collaborative project, with the aim of comparing adolescent quality of life (QOL) internationally. This study presents the primary results of the investigation conducted in Beijing, China, and Kobe, Japan. The 70-item Quality of Life Questionnaire for Adolescents (QOLQA) was developed and evaluated in Japan and China. In total, 1114 Japanese and 613 Chinese junior high school students, aged 12-15 years, completed the questionnaire. Chinese students scored significantly higher than the Japanese students in overall QOL scores and in most domains. For both groups, subjects had highest score in the independence domain and lowest in psychological domain. In terms of overall QOL score, Chinese male students ranked first, followed by Chinese girls, Japanese boys, and Japanese girls. In the Japanese group, a continuing decrease of QOL scores with age was observed without exception, but no such tendency was present in their Chinese counterparts. No parallel relationship was observed between the higher level of economic development and better quality of life. The results also suggest that mental health promotion should be a priority in improving overall quality of life of adolescents both in Japan and China. PMID- 10803808 TI - Age at onset of schizophrenia: gender differences and influence of temporal socioeconomic change. AB - This study was undertaken to examine whether males develop schizophrenia at a younger age than females, and whether temporal socioeconomic change affects the age at onset of schizophrenia. The subjects were 848 ICD-9 schizophrenics who were admitted to Nihon University Hospital, Tokyo, Japan, during the period of 1955-64 (n = 468 (214 males and 254 females), group A) or during the period of 1982-91 (n = 380 (220 males and 160 females). group B). Schizophrenic males showed an earlier age at onset than schizophrenic females. However, the mean age at onset of schizophrenia did not differ significantly between group A and group B. These results indicate that the gender difference in age at onset of schizophrenia has not been influenced by temporal socioeconomic change. PMID- 10803809 TI - Hyperperfusion in the lateral temporal cortex, the striatum and the thalamus during complex visual hallucinations: single photon emission computed tomography findings in patients with Charles Bonnet syndrome. AB - We report the findings of single photon emission computed tomography using 123I IMP and magnetic resonance image studies of five patients with Charles Bonnet syndrome (CBS) while they were having visual hallucinations. All patients developed complex visual hallucinations after suffering from eye disease. The mean age at onset of CBS was 71.6 years. Single photon emission computed tomography studies in all patients disclosed hyperperfusion areas with some asymmetrical appearances in the lateral temporal cortex, striatum and thalamus. These results suggest that when elderly people suffer from eye disease, subsequent excessive cortical compensation in the lateral temporal cortex, striatum and thalamus may precipitate the development of visual hallucinations. PMID- 10803810 TI - Watershed infarction associated with dementia and cerebral atrophy. AB - A 63-year-old man was admitted with progressive left hemiparesis and left homonymous hemianopsia of 1 month's duration. During the 2 months before admission, he had suffered from slowly progressive dementia. The diagnosis of right-sided watershed (WS) infarction was made. He exhibited slow progression of dementia and cerebral atrophy during the period of observation after discharge. There was a positive relationship between cerebral atrophy and the degree of dementia. In the present case, WS infarction caused by right internal carotid artery occlusion might be related to dementia and cerebral atrophy. PMID- 10803811 TI - Does smoking cessation improve mental health? AB - There is a possibility that cessation of smoking improves mental health, but there are no studies that have demonstrated this. A cohort study was performed for 1 year in 18 males who spontaneously stopped smoking (cessation group) and 173 who continued to smoke (smoking group). The mental health state was evaluated using the Japanese version of the 30-item General Health Questionnaire (GHQ-30) before the cessation of smoking and 6 months and 1 year after smoking cessation. Changes in the GHQ score were compared between the cessation and smoking groups. In order to control the effects of confounding factors, multiple regression analyses were performed using the GHQ score after 6 months and 1 year as dependent variables. The GHQ score in the cessation group significantly decreased 6 months and 1 year after smoking cessation (P < 0.04 and 0.01, respectively, by paired t-test). In the smoking group, the GHQ score slightly decreased. Repeated measure analysis of variance revealed that the decrease in the GHQ score in the cessation group was significantly larger than in the smoking group. Multiple regression analysis revealed significant effects of smoking cessation on mental health after controlling for other confounding factors. It can be concluded that smoking cessation may improve mental health. PMID- 10803812 TI - A clinical study on suicide among schizophrenics. AB - We conducted a clinical investigation of 80 suicides who met the DSM-III-R criteria for schizophrenia. The results using this approach showed no significant difference with those of preceding studies, and general features regarding the phenomenology of suicide among schizophrenics worldwide were recognized. The present study, however, served to underscore the following points: (i) suicide of schizophrenics must be considered a concern at all stages of the disease; (ii) the subjective strength of the will to die may be more important for the committing suicides than the lethality of the methods employed; and (iii) a change in the environment, for example, a hospital admission or discharge, may trigger suicide. A control group of 80 living schizophrenics with no past attempted suicide was then matched to the suicide group with respect to sex and illness duration, in order to identify the predictors of suicide. In a logistic regression analysis, the presence of suicidal ideation, degree of anxiety estimated by positive and negative syndrome scale, and birth order were revealed as predictors of suicide. As to the birth order, the risk is higher in middle children. PMID- 10803813 TI - Typus melancholicus and the Temperament and Character Inventory personality dimensions in patients with major depression. AB - Although many clinical studies have been conducted to determine the etiological role and clinical implications of typus melancholicus for unipolar depression, maladaptive personality features in depressive patients have not been well described. This study explores typus melancholicus, as measured by the rigidity subscale of the Munich Personality Test, and maladaptive personality features, as measured by the Temperament and Character Inventory (TCI), in 131 remitted patients with DSM-IV major depression and 154 normal controls. The patients reported significantly higher scores on rigidity and harm avoidance and significantly lower scores on self-directedness and cooperativeness. Only 23.6% of the variance of the rigidity scale was explained by the variance of the seven TCI scales, in which only persistence was significantly correlated positively to rigidity. Cluster analysis identified four subgroups, two of which were characterized by a high rigidity score. One of these two subgroups showed no maladaptive personality features, as measured by the TCI, while the other showed high harm avoidance and low self-directedness. These results indicate that the personality of depressive patients is characterized not only by typus melancholicus but also by maladaptive personality features, that typus melancholicus is not well represented by any TCI scale, and that typus melancholicus and maladaptive personality features can coexist in some depressive patients. PMID- 10803814 TI - Functional significance of stimulatory GTP-binding protein in hippocampus is associated with kindling-elicited epileptogenesis. AB - In order to evaluate the involvement of the stimulatory G-protein (Gs)-related transduction system in the basic mechanisms of epilepsy, we examine the expression levels of Gsalpha mRNA and specific GTP-binding ability in the hippocampus of amygdaloid-kindled rats at various seizure stages. Northern blot analysis showed a significant increase in the Gsalpha mRNA expression level in the bilateral hippocampus at 24h after the last generalized seizure. The [3H]-GTP binding assay with isoproterenol (IPN), a beta-receptor agonist, revealed a remarkable increase of Bmax values in the sham-operated control and partially kindled groups. However, the IPN-induced increase of Bmax values was abolished on both sides of the hippocampus at 24 h after and at 4 weeks after the last generalized seizure in fully kindled rats. These data suggest that alteration in the Gs function and beta-adrenergic receptor-Gs coupling might be implicated in the neurobiological basis of the induction mechanisms of the generalization of seizures and the mechanisms of the maintenance of enduring epileptogenesis. Conversely, the Gs-related transduction system might have a lesser impact on the acquisition process of epileptogenesis. PMID- 10803815 TI - Tau protein is a potential biological marker for normal pressure hydrocephalus. AB - A biological marker for normal pressure hydrocephalus (NPH) is beneficial for evaluation of its severity and of indications for shunt operation. Tau protein was initially considered as a biological marker in cerebrospinal fluid (CSF) from Alzheimer's patients. Recently, it has been demonstrated that degeneration in the brain causes elevation of tau in CSF. Therefore, the tau level in CSF from NPH patients was evaluated. Tau levels in CSF from NPH patients were significantly higher than that in controls. The tau levels were correlated with the severity of dementia, urinary incontinence, and gait disturbance in NPH. These results suggest that CSF tau may be useful as a biological marker for NPH to determine the level of neuronal degeneration. PMID- 10803816 TI - Obsession of hearing music: from the viewpoint of Morita theory. AB - We present two typical cases among five Japanese students who have complained of hearing music. They were found among 638 student case reports during 4 years at the mental health service center of a preparatory school. We diagnosed the complaint of hearing music as a symptom of obsession. Morita therapy was an effective treatment for this symptom. We discuss the characteristics of the symptom as an obsession and the therapy in the light of Morita theory. PMID- 10803817 TI - A short daytime nap modulates levels of emotions objectively evaluated by the emotion spectrum analysis method. AB - A novel objective technique, the emotion spectrum analysis method, was first applied to investigate emotional fluctuations before, during and after a daytime nap in eight healthy young adults (four males and four females). The subjects were allowed to freely nap between 13.00 and 14.00 h, in which stages 1 and 2 non rapid eye movement sleep occurred on average for 5.9 and 20.8 min, respectively. Emotional components such as anger, joy, relaxation and sadness were numerically estimated on the basis of spatio-temporal behavior of 21-channel electroencephalogram and analyzed statistically. In comparison with the prenap waking level, the magnitudes of the anger, joy and relaxation components remained stably unchanged during the nap but elevated significantly during the postnap waking period. The sadness component exhibited little significant change throughout the observation period. From the results, we suggest that a short daytime nap modulates the emotions to improve the postnap mental states. PMID- 10803818 TI - Factors associated with admission to a geriatric hospital in semisuburban southern Japan. AB - A case-control study was conducted to evaluate the factors associated with admission to a geriatric hospital. Case studied were 13 Japanese elderly who were admitted to a geriatric hospital because their female caregivers had found it impossible to look after them at home. We used 35 pairs of elderly and female caregivers, who were receiving domiciliary visits by nurses, in the catchment area of the hospital. The present study revealed that elderly with dementia (vs without; Odds ratio = 6.69) and with moderately limited activities of daily living (Barthel Index 61 +) (vs severely limited activities of daily living: Barthel Index 0-60; Odds ratio = 6.62), caregivers being a daughter-in-law (vs other kinship; Odds ratio = 6.30), were risk factors. PMID- 10803819 TI - Declining of memory functions of normal elderly persons. AB - Two studies examined the declining of memory functions in normal elderly persons using the Yokota memory test (YMT), which includes 15 items concerning verbal and non-verbal memory functions. In the first study, 552 subjects over 40 years of age in five age groups were examined. Factor analysis revealed that YMT consisted of two factors pertaining to short-term/working memory, and two factors pertaining to long-term memory. It is suggested that the former was more affected than the latter, with aging. In the second study, YMT was examined in relation to the revised version of Hasegawa dementia scale (HDS-R), which was the most popular intelligence scale for the elderly in Japan. As a result, memory functions differentially declined with the decreasing score of HDS-R, which suggests that memory functions differentially declined with progressive risk of dementia. PMID- 10803820 TI - The central mechanism of hypothalamic-pituitary-adrenocortical system hyperfunction in depressed patients. AB - While hypercortisolemia is commonly observed in depression, exactly where in the hypothalamic-pituitary-adrenocortical (H-P-A) axis this dysfunction arises remains undefined. In attempting to distinguish between central or peripheral locus of dysfunction, we studied in 12 patients (10 females, two males) with primary major depression and eight age-matched controls (six females, two males) in their adrenal cortisol response to infused adrenocorticotropic hormone (ACTH) (cosyntropin 0.05 microg/kg bodyweight) while endogenous ACTH was suppressed with 1 mg of dexamethasone. Compared with the control group, pre-dexamethasone plasma baseline cortisol level was significantly higher in depressed patients while ACTH level remained normal. Post-dexamethasone responses of both hormones were greatly non-suppressed in the depressed group. Exogenous cosyntropin-elicited rise in plasma cortisol was significantly lower in depressed patients while the ACTH response was not significantly different. These findings suggest that an adrenal cortisol response to ACTH was significantly decreased during depression as compared with normals in Chinese depressed patients. Therefore, the central mechanism of hyperfunctioning H-P-A axis causing hypercortisolemia should be emphasized. PMID- 10803821 TI - Affective communication of infants with autistic spectrum disorder and internal representation of their mothers. AB - We have been examining the developmental process of affective communication in infants with autistic spectrum disorders from the viewpoint of relationship disturbance through our developmental and psychopathological studies on autism. In particular, the role of internal representation of the mother in the process of development of affective communication is discussed through the presentation of two cases diagnosed as autistic spectrum disorder in early infancy. In these cases, we postulate approach-avoidance motivational conflict as the primary factor impeding development of affective communication, focusing therapeutic intervention on this perspective. As a result, attachment behavior was remarkably improved in the children, but affective communication with their mothers was not readily improved. Taking up the mothers' own internal representation in mother infant psychotherapy, in particular, the mothers' problems in attachment behavior with their own mothers in infancy precipitated transition in the mothers' internal representation of their children, leading to active evolution in mother child interaction and development in affective communication between mother and child. In this context, the basis and significance of internal representation of both parties being determinants in the quality of mother-child communication are discussed. PMID- 10803822 TI - Reliability of clinical judgment of patients' competency to give informed consent: a case vignette study. AB - The competency of psychiatric patients to give informed consent is important in respecting patients' decisions as well as protecting patients from undue exploitation. A total of 176 members of the Japanese Society of Psychiatry and Neurology gave a clinical judgment in a questionnaire of competency in five case transcriptions. Their interrater reliability of competency judgment was slight (generalized kappa 0.31). Clinicians' global judgment of patients' competency was not reliable, but it may be improved by the use of a structured interview. PMID- 10803823 TI - Patient satisfaction with polypharmacy reduction in chronic epileptics. AB - The effects of polypharmacy reduction on patient satisfaction and subjective seizure severity were assessed prospectively in adult out-patients with chronic epilepsy using Japanese versions of the Side effects and Life Satisfaction (SEALS) and the Seizure Severity Questionnaires (SSQ). Antiepileptic drugs (AED) were withdrawn using a 1-year reduction schedule. The SSQ score was not aggravated and total SEALS score improved significantly. Moreover, temper subscore was also improved in the sedative AED reduction group. Similar to previous studies from the physician's viewpoint, the present study confirms that from the perspective of the patient, polypharmacy reduction, especially withdrawal of sedative AED, has a favorable effect on patient satisfaction. PMID- 10803824 TI - Therapeutic implications of the new biology. PMID- 10803825 TI - Sentinel lymph node biopsy for the detection of metastases. AB - Although the concept of the sentinel node has existed for most of the twentieth century, it was only in the last decade that methods were developed which allowed practical application to cancer patients. Sentinel nodes (SN) are a highly variable but limited set of lymph nodes first to receive drainage from any given location. Cancer metastasizes to these nodes before other nodes. Radioactive tracers and blue dyes are used to guide the surgeon to the SNs for resection. Two large clinical trials of SN surgery in breast cancer patients are actively underway in the United States. Until the impact of this very promising technology is confirmed to provide the same staging and prognosis, regional control, and survival as conventional surgery, it is considered an experimental procedure. PMID- 10803826 TI - High-dose chemotherapy with peripheral blood progenitor cell transplantation in the adjuvant treatment of breast cancer. AB - High-dose chemotherapy as adjuvant treatment for high-risk breast cancer has been and is being investigated in a series of randomized trials. The preliminary information from 5 studies is difficult to interpret. The data are consistent with a modest benefit of high-dose therapy, but strong evidence for this is lacking. Additional follow-up and the results of several large trials outside the United States are awaited. Hitherto unrecognized pharmacologic interactions between high-dose agents and long-term toxicities such as cognitive function impairment complicate the issue considerably. PMID- 10803827 TI - The clinical significance of tumor progression: breast cancer as a model. PMID- 10803828 TI - Angiogenesis and cancer metastasis. AB - The growth and spread of neoplasms depends on the establishment of an adequate blood supply, that is, angiogenesis. The onset of angiogenesis involves a change in the local equilibrium between proangiogenic and antiangiogenic regulators that are produced by tumor cells, surrounding stromal cells, and infiltrating leukocytes. In most normal tissues, factors that inhibit angiogenesis predominate, whereas in rapidly dividing tissues, the balance of angiogenic molecules favors stimulation of the process. A potent inhibitor of angiogenesis is interferon-alpha or -beta, shown to down-regulate transcription and protein production of basic fibroblast growth factor, collagenase type IV, and interleukin-8. The daily systemic administration of low (but not high) dose of interferon-alpha can produce significant inhibition of angiogenesis and, hence, regression of human tumors implanted orthotopically in nude mice. The recent elucidation of the interaction among proangiogenic molecules during physiological processes and the apparent disruption of this balance in neoplasia should allow the design of potent antiangiogenic therapies against primary cancers and metastases. PMID- 10803829 TI - The identification of cancer antigens: impact on the development of cancer vaccines. PMID- 10803830 TI - Hodgkin's disease. AB - There is overwhelming evidence that at least a substantial number of cases, if not all, of Hodgkin's disease (HD) represent monoclonal B-cell disorders. The treatment of HD is changing strikingly. In early stages of disease, extended field irradiation has been the standard resulting in excellent cure rates. Due to the recognition of fatal long-term effects, however, especially the high rates of second solid tumors, extended field radiotherapy is now being abandoned by most study groups. Instead, mild chemotherapy for control of occult disease is combined with involved field irradiation. In intermediate stage HD, where combined modality treatment is the treatment of choice, extended field irradiation is substituted by involved field irradiation for the same reasons. In advanced stage HD, 8 cycles of polychemotherapy (plus additional radiotherapy for large tumor masses and residual lymphomas) for decades has cured only 50% to 60% of patients. The development of a new dose-intensified regimen (BEACOPP) for the first time has significantly improved that prognosis. In relapsed HD, recently published phase III studies suggest an improvement of relapse-free survival of patients using high-dose chemotherapy followed by autologous stem cell transplantation (ASCT). PMID- 10803831 TI - The role of laparoscopy in the treatment of intra-abdominal malignancies. AB - The role of laparoscopy in the care of patients with cancer is currently evolving. Numerous experimental and clinical studies have attempted to elucidate the nature and cause of port-site metastases--particularly to discern whether they simply are a marker of advanced disease, or if they are a result of the laparoscopic intervention. Laparoscopy has a role in establishing the diagnosis of cancer in some situations by allowing biopsy of intraperitoneal and retroperitoneal masses, lymph nodes, and visceral lesions, as well as examination of abdominal contents under direct vision or with ultrasound probes. Laparoscopy is useful in the staging of established malignancies such as pancreatic cancer, hepatic lesions, lymphoma, and esophageal cancer. Laparoscopy also has a role in the surgical treatment of a variety of malignancies, including gastric carcinoma, pancreatic cancer, splenic malignancies, adrenal cancers, and colon cancer. The safety of laparoscopy for the definitive resection of colon cancer has not yet been proven, and until the results of a randomized prospective trial currently underway are known, should be performed only in the context of a clinical trial. Lastly, laparoscopy can play an important role in the palliative care of the cancer patient in performing procedures such as feeding-tube placement or intestinal stoma creation. It is imperative that using laparoscopy in the care of patients with malignancies be carefully and thoroughly evaluated since this technique can either benefit or adversely affect survival or quality of life. PMID- 10803832 TI - Treatment of patients with hepatic metastases. PMID- 10803833 TI - Multimodality therapy of esophageal cancer: an update. AB - The incidence of adenocarcinoma of the esophagus is rapidly increasing in Western countries, particularly in Caucasian men. Both squamous cell carcinoma and adenocarcinoma are equally virulent tumors. In the absence of effective prevention and screening programs, most Western patients are found to have locally advanced or metastatic disease at the time of diagnosis. The outcome with regional therapy alone (either surgery or radiation) is poor. Multimodality therapy--treatment plans which include both systemic and regional treatments-is the focus of intense study. This review will summarize the current status of several different approaches that have the goal of increasing the cure rate in patients with newly diagnosed esophageal cancers. PMID- 10803834 TI - Screening and hormonal therapy of localized prostate cancer shows major benefits on survival. AB - Five recent randomized studies demonstrate that treatment of localized prostate cancer saves lives. Since prostate cancer grows insidiously without signs or symptoms until it reaches the bones-when cure has become an exception-screening is essential to diagnose the disease at an earlier stage when cure is possible in the majority of patients. At 7 years' median follow-up with continuous hormonal therapy, cancer-specific death was observed in less than 5% of patients with treatment started at the stage of clinically localized disease, compared to 70% to 90% when treatment is started in patients with bone metastases. With appropriate screening, 99% of prostate cancers can be diagnosed at the localized stage with no sign of bone metastases. Starting androgen blockade at time of diagnosis of localized disease can achieve long-term control of the disease in close to 100% of patients and even cure the disease in the majority of them, as evidenced by maintenance of undetectable PSA in 85% of patients after cessation of long-term combined hormonal therapy. The available data indicate that the use of regular PSA screening and immediate treatment can markedly reduce mortality rates among prostate cancer patients. Next steps toward improving the quality of life for prostate cancer patients including more tolerable treatments and prevention steps are underway. PMID- 10803835 TI - Prostate specific antigen: a useful screening test? AB - The role of prostate specific antigen (PSA) as a screening test for prostate cancer is controversial. Proponents of screening emphasize that early detection can lead to discovery of organ-confined disease and the potential for cure. Opponents point to the lack of credible evidence that screening is associated with a decrease in mortality. In addition, population-based screening (with subsequent diagnosis and treatment in many men) can be associated with considerable morbidity and mortality in the context of a disease that is often not fatal. This report examines the limitations of PSA as a screening test and supports an approach using "verbal informed consent" to identify patients who should be tested. PMID- 10803836 TI - Failure pattern implications following external beam irradiation of prostate cancer: long-term follow-up and indications of cure. AB - The purpose of this study was to present patterns and risk of biochemical failure following external beam irradiation of prostate cancer and to make comparisons to a published modern radical prostatectomy series. Between January 1987 and December 1994, 328 men were treated definitively at Fox Chase Cancer Center for localized prostate cancer using conventional or three-dimensional conformal radiotherapy. The median biochemical follow-up was 6.4 years, with all patients having at least 5 years follow-up. Two prognostic patient groups were established on the basis of proportional hazards modeling that considered treatment and presenting tumor characteristics. For each of the two prognostic groups, biochemical failure and hazard functions were estimated using the ASTRO consensus definition of failure and life table methodology. Failure risk comparisons were made to modern published radical prostatectomy series. Multivariate analysis demonstrated the independent predictive power of pretreatment PSA level, palpation stage, Gleason score, and dose. Thus, the favorable prognosis group, Group I, consisted of 83 patients who were treated with a dose level > or = 74 Gy and who presented with PSA levels < 20 ng/ml, T1/T2A tumors, and Gleason score 2 6. Group II consisted of 245 patients with at least one of the following: pretreatment PSA level > or = 20 ng/ml, T2B/T3 tumor, Gleason score 7-10, dose < 74 Gy. The 5- and 8-year bNED estimates were 76% and 76% for Group I, and 51% and 49% for Group II. Only three failures occurred after 5 years, all from Group II, representing 2% of the total failures observed. Hazard function estimates indicate maximum risk of failure at 24 to 36 months, tapering to a low rate at 4 years with no failures observed after 6 years. Differences in patterns of failure by prognostic group show maximum risk of failure at 24 months (median, 31 months) for Group I, and 12 to 36 months (median, 22 months) for Group II. Group II reaches low levels of risk at 6 years, in contrast to 4 years for the patients with a more favorable prognosis. We concluded that patients treated with external beam radiation alone show little risk of failure after 4 to 6 years. This result suggests that the 5-year bNED control rate approximates the eventual cure rate of prostate cancer. PMID- 10803837 TI - A novel HLA-A*6816 allele possible generated by a point mutation in a Chilean from Punta Arenas (Magellan Strait). PMID- 10803838 TI - Founder effect for a 26-bp deletion in the RFXANK gene in North African major histocompatibility complex class II-deficient patients belonging to complementation group B. AB - Expression of major histocompatibility complex (MHC) class II genes is controlled at the transcriptional level by at least four trans-acting genes, CIITA, RFXANK, RFX5, and RFXAP. Defects in these regulatory genes result in the absence of MHC class II molecule expression and, thereby, cause a combined immunodeficiency. MHC class II deficiency is inherited as an autosomal recessive trait. Since the first description of the disease, about 70 patients from 50 families have been reported. Forty-three of these families have been classified into four complementation groups: A, B, C, and D. In the largest group, B, the majority of patients are of North African origin. In two of these patients, the same mutation in the RFXANK gene (752delG-25) was identified. We performed a mutation analysis in 20 additional patients belonging to complementation group B and detected the 752delG-25 mutation in 17. All of these patients are of North African origin. A founder effect for this mutation was documented, since all tested patients, except one, display a common haplotype spanning the RFXANK locus. PMID- 10803839 TI - The genomic organization and evolution of the natural killer immunoglobulin-like receptor (KIR) gene cluster. AB - Natural killer (NK) immunoglobulin-like receptors (KIRs) are a family of polymorphic receptors which interact with specific motifs on HLA class I molecules and modulate NK cytolytic activity. In this study, we analyzed a recently sequenced subgenomic region on chromosome 19q13.4 containing eight members of the KIR receptor repertoire. Six members are clustered within a 100-kb continuous sequence. These genes include a previously unpublished member of the KIR gene family 2DS6, as well as 2DL1, 2DL4, 3DL1, 2DS4, 3DL2, from centromere to telomere. Two additional KIR genes, KIRCI and 2DL3, which may be located centromeric of this cluster were also analyzed. We show that the KIR genes have undergone repeated gene duplications. Diversification between the genes has occurred postduplication primarily as a result of retroelement indels and gene truncation. Using pre- and postduplication Alu sequences identified within these genes as evolutionary molecular clocks, the evolution and duplication of this gene cluster is estimated to have occurred 30-45 million years ago, during primate evolution. A proposed model of the duplication history of the KIR gene family leading to their present organization is presented. PMID- 10803840 TI - CC-chemokine receptor 5 polymorphism and age of onset in familial multiple sclerosis. Multiple Sclerosis Genetics Group. AB - Multiple sclerosis (MS) is a common disease of the central nervous system characterized by myelin loss and progressive neurological dysfunction. An underlying genetic susceptibility plays a clear role in the etiology of MS, likely acting in concert with an undefined environmental exposure. Full-genome screenings in multiplex MS families have identified several susceptibility regions, supporting a polygenic model for MS. Among these regions, evidence for weak linkage was observed at 3p/3cen suggesting the presence of an MS gene(s) of modest effect. Encoded here are two chemokine receptors, CCR5 and CCR2B. We examined the chromosome 3p21-24 region in 125 MS families (322 total affecteds and 200 affected sib-pairs), and performed genetic analyses of CCR5 and CCR2B loci and two nearby markers (D3S1289 and D3S1300) using both linkage- and association-based tests. No evidence of linkage to MS was observed for any of the tested markers. Affected relative-pair (SimIBD) and sib-pair analyses (ASPEX), and association testing (sib-TDT) for each locus were also not significant. However, age of onset was approximately 3 years later in patients carrying the CCR5delta32 deletion (P=0.018 after controlling for gender effects). Thus, chemokine receptor expression may be associated with differential disease onset in a subset of patients, and may provide a therapeutic target to modulate inflammatory demyelination. PMID- 10803841 TI - Role of nuclear factor-kappaB in the expression by tumor necrosis factor-alpha of the human polymeric immunoglobulin receptor (plgR) gene. AB - We analyzed the mechanism of human polymeric immunoglobulin receptor (pIgR) gene upregulation by tumor necrosis factor (TNF)-alpha. Northern blot analysis showed that the expression of pIgR mRNA was enhanced by TNF-alpha stimulation. This activation was completely inhibited by RNA polymerase or protein synthesis inhibitors, suggesting that the regulation of pIgR gene expression depends on de novo RNA and protein synthesis. Furthermore, the stimulation of pIgR mRNA by TNF alpha was decreased by pyrrolidinedithiocarbamate and L-1-4'-tosylamino phenylethyl-chloromethyl ketone, which are known nuclear factor (NF)-kappaB inhibitors. For further analysis of gene regulation, we cloned and sequenced the 1.5-kb 5'-flanking region of the pIgR gene. In the upstream region, we found two NF-kappaB-binding motifs (named kappaB1 and kappaB2 from the 5' region). An electrophoretic mobility shift assay indicated that two components of the NF kappaB/Re1 family, p50 and p65, bound with higher affinity to the KB2 element than to the kappaB1 element. We also analyzed plgR gene expression using reporter plasmids expressing the firefly luciferase gene. Stimulation by TNF-alpha significantly activated the pIgR gene promoter, as a 775-bp upstream region of the pIgR gene increased luciferase gene expression in cells treated with TNF alpha. The activation of promoter activity by TNF-alpha was abolished when a mutation was inserted into kappaB1 or kappaB2. These data indicated that pIgR gene expression induced by TNF-alpha is transcriptionally regulated via activation of NF-kappaB. In addition, there is a possibility that another factor may act in concert with NF-kappaB. PMID- 10803842 TI - A polymorphism of the rat T-cell receptor beta-chain variable gene 13 (BV13S1) correlates with the frequency of BV13S1-positive CD4 cells. AB - Three rat BV13S1 alleles (T-cell receptor beta-chain variable gene 13) were characterized by new BV13S1-allele specific monoclonal antibodies (18B1 and 17D5) and sequence analysis of expressed and genomic BV13S1. Two alleles were functional and designated BV13S1A1 present in strains LEW, BUF, PVG, and BV13S1A2 present in BN and WF. Their products differed by six amino acids, two of them in complementarity-determing region (CDR)1 and one in CDR2. A third nonfunctional allele, BV13S1A3P, was found in strains F344 and DA. Apart from a single nucleotide insertion, it was identical to BV13S1A2. All 12 rat strains tested showed association of TCRBC1 with BV8S2/4 alleles but not with the BV13S1 alleles, which may reflect a different gene order of the rat BV compared to mouse. BV13S1A1-encoded T-cell receptors (TCRs) which bind both monoclonal antibody (mAb) 18B1 and mAb 17D5 are over-represented in the CD4 lymphocyte subset. BV13S1A2-encoded TCRs which are stained by mAb 18B1 but not by mAb 17D5 show a slight CD8-biased expression. Preferential usage of BV13S1A1-positive TCRs by CD4 but not by CD8 cells in (LEW x WF)F1 hybrids and cosegregation of BV13SA1 and increased frequency of BV13S1 TCR-positive CD4 cells in a (LEW x BN)x BN backcross suggest structural differences of the two allelic products as the reason for their contrasting CD4/CD8 subset bias. PMID- 10803843 TI - Molecular cloning of transmembrane and soluble forms of a novel rat natural killer cell receptor related to 2B4. AB - Natural killer (NK)-cell recognition of target cells and cytolytic function are controlled by multiple receptor-ligand interactions. These receptors can transmit either positive or negative signals and belong to the lectin superfamily or immunoglobulin superfamily (IgSF). One member of the IgSF, 2B4, is expressed on the surface of all mouse and human NK cells and the subset of T cells that mediate NK-like killing. In both mouse and human, 2B4 is a transmembrane protein and is the counter-receptor for CD48. Northern blot analysis had indicated the existence of 2B4-related genes. Here we report the cloning of novel cDNAs (r2B4R) closely related to the rat 2B4. Unlike 2B4, rat NK cells express mRNA corresponding to both transmembrane (r2B4R-tm) and soluble (r2B4R-se) forms of r2B4R. r2B4R-tm contains an open reading frame encoding a polypeptide of 311 amino acid residues. The encoded protein has characteristics of type I transmembrane proteins with a 20-amino acid leader sequence, a 203-amino acid extracellular domain, a 23-amino acid transmembrane domain, and a 65-amino acid cytoplasmic domain. r2B4R-se encodes a protein of 205 amino acid residues without a putative transmembrane domain. Northern blot analysis and reverse transcriptase PCR analysis revealed that both transmembrane and soluble forms of r2B4R are expressed in interleukin-2-activated NK cells. PMID- 10803844 TI - A high incidence of Shigella-induced arthritis in a primate species: major histocompatibility complex class I molecules associated with resistance and susceptibility, and their relationship to HLA-B27. AB - The human major histocompatibility complex (MHC) class I gene, HLA-B27, is a strong risk factor for susceptibility to a group of disorders termed spondyloarthropathies. Rodents that express HLA-B27 develop spondyloarthropathies, implicating HLA-B27 in the etiology of these disorders. To determine whether an HLA-B27-like molecule was associated with spondyloarthropathies in nonhuman primates, we analyzed the MHC class I cDNAs expressed in a cohort of rhesus macaques that developed reactive arthritis after an outbreak of shigellosis. We identified several cDNAs with only limited sequence similarity to HLA-B27. Interestingly, one of these MHC molecules had a B pocket identical to that of HLA-B39. Pool sequencing of radiolabeled peptides bound by this molecule demonstrated that, like HLA-B27 and HLA-B39, it could bind peptides with arginine at the second position. However, extensive analysis of the MHC class I molecules in this cohort revealed no statistically significant association between any particular MHC class I allele and susceptibility to reactive arthritis. Furthermore, none of the rhesus MHC class I molecules bore a strong resemblance to HLA-B27, indicating that reactive arthritis can develop in this animal model in the absence of an HLA-B27-like molecule. Surprisingly, there was a statistically significant association between the rhesus macaque MHC A locus allele, Mamu-A*12, and the absence of reactive arthritis following Shigella infection. PMID- 10803845 TI - Characterization of the Ly49I promoter. AB - Fourteen potential Ly49 genes have been identified in the C57B1/6 mouse strain, and cDNAs containing a complete coding region have been isolated for 10 members of this gene family. Ly49 proteins are primarily expressed in natural killer (NK) cells. Although the sequence of the Ly49a promoter region has been published, no study of the cell-specific activity of the promoter has been reported. A 12-kb genomic fragment of the Ly49I gene was isolated and characterized by DNA sequencing. Approximately 5 kb of DNA sequence upstream of the first Ly49I exon was determined and this region was used to perform promoter analysis using luciferase reporter plasmid constructs. A core promoter was identified that was preferentially transcribed in a Ly49-expressing cell line, EL-4. Electrophoretic mobility shift assays using oligonucleotide probes from the core Ly49i promoter and comparable regions from the Ly49a promoter demonstrated the importance of TATA-related elements in generating EL-4 and NK cell-specific DNA/protein complexes. PMID- 10803846 TI - Molecular cloning of the gene for interleukin-1beta from Xenopus laevis and analysis of expression in vivo and in vitro. AB - The Xenopus cDNA for interleukin-1beta (IL-1B) was cloned and sequenced. The gene contains 1,462 nucleotides that translate in a single reading frame to give a predicted 283-amino acid IL-1beta molecule. The translated molecule contains a single potential glycosylation site, a readily identifiable IL-1 family signature, and has highest homology to chicken IL-1beta by phylogenetic tree analysis and sequence similarity. It lacks a signal peptide in common with other known IL-1B genes, and lacks a clear ICE (caspase) cut site in common with other nonmammalian IL-1B genes sequenced to date. RT-PCR was used to study sites of IL 1B transcript expression, 24 h following injection of lipopolysaccharide (LPS). Expression was detected in the brain, liver, kidney, and spleen, with expression weakest in the brain and strongest in the spleen. No transcript expression was detectable following injection of saline. Northern blot analysis was used to quantify the induction of IL-1B expression in splenocytes following in vivo or in vitro stimulation with LPS. The results are discussed in relation to the potential role of IL-1beta in amphibian immune responses. PMID- 10803847 TI - Molecular and functional characterization of a fish inducible-type nitric oxide synthase. AB - Using an oligonucleotide primer based on a partial goldfish inducible nitric oxide synthase (iNOS) sequence, a complete carp iNOS cDNA was isolated from an activated carp phagocyte cDNA library. Nucleotide and predicted amino acid sequence analysis indicate that carp iNOS encodes a 1,127-amino acid protein with 57% sequence identity to human iNOS. Like mammalian NOSs, carp iNOS protein contains putative binding sites for heme, tetrahydrobiopterin, calmodulin, flavine mononucleotide, flavine adenine dinucleotide, and NADPH. Phylogenetic analysis, using neighbor joining, showed that the carp iNOS protein clustered together with the other vertebrate iNOS proteins. Inducibility of carp iNOS was confirmed by reverse transcription-polymerase chain reaction after stimulation of carp phagocytes with lipopolysaccharide or the protozoan blood flagellate Trypanoplasma borreli. These stimulators produced high amounts of nitric oxide that were toxic for T. borreli in vitro. The nuclear transcription factor NF kappaB was shown to play a role in the induction of iNOS transcription. PMID- 10803848 TI - Molecular characterization of a new member of the immunoglobulin superfamily that potentially functions in T-cell activation and development. AB - Differential hybridization cloning has been used to isolate a novel chicken thymic activation and developmental sequence (cTADS). The nucleotide sequence of the cTADS cDNA predicts an open reading frame of 439 amino acids. The inferred cTADS protein possesses a hydrophobic membrane-spanning domain and putative intracellular kinase activation domains. Its extracellular domain shares similarities with the immunoglobulin protein superfamily, featuring two conserved immunoglobulin folds that resemble C1 and C2 constant regions. The cTADS sequence shows similarity to a subfamily of proteins involved in cellular adhesion: chicken neural cell adhesion molecule and human opioid-binding adhesion molecule, and to proteins that have a biological role in intracellular signaling: mouse platelet-derived growth factor receptor and human fibroblast growth factor receptor. cTADS is differentially expressed in chicken thymic cells during embryonic development and during activation through the T-cell receptor. Sequence similarities and expression patterns suggest that cTADS could be involved in cell recognition and adhesion, and/or peptide ligand binding. PMID- 10803849 TI - Cloning, chromosomal location, and tissue expression of the gene for pig interleukin-18. AB - Interleukin 18 (IL-18), a recently described cytokine, plays an important role in the cell-mediated immune response, in particular through its ability to induce the production of interferon (IFN)-gamma. We cloned pig IL-18 cDNA from the intestinal epithelial cell line IPI-2I using a reverse transcriptase-polymerase chain reaction method with primers derived from the human IL-18 sequence. The amino acid sequence deduced from pig IL-18 cDNA encodes a 192 amino-acid polypeptide that exhibits 92, 90, 81, and 71% similarity to IL-18 from horse, dog, human, and rodents (mouse and rat), respectively. Structural comparison of the IL-18 protein with IL-1alpha and IL-1beta showed that IL-18 shares several characteristics with the IL-1 cytokine family: the IL-1 signature-like sequence, a potential caspase-1 (ICE) cleavage site, and the presence of 12 predicted beta strands. Fluorescence in situ hybridization was used to localize the IL-18 gene on the short arm (p13) of pig chromosome 9. Analysis of IL-18 expression in different organs of piglets demonstrated that IL-18 mRNA is weakly expressed in the kidney and the lung. By contrast, we observed highly constitutive expression of IL-18 mRNA in the spleen, mesenteric lymph nodes, and the intestine, particularly in the small intestine, indicating a potential role for IL-18 as a first line of host defense in the intestinal mucosa. PMID- 10803850 TI - Molecular basis for factor H and FHL-1 deficiency in an Italian family. PMID- 10803851 TI - Lupus susceptibility loci map within regions of BXSB derived from the SB/Le parental strain. PMID- 10803852 TI - BTL-II: a polymorphic locus with homology to the butyrophilin gene family, located at the border of the major histocompatibility complex class II and class III regions in human and mouse. AB - Comparison of human and mouse genomic sequence at the border of the major histocompatibility complex (MHC) class II and class III regions revealed a locus encoding six exons with homology to the butyrophilin gene family and the location of a previously described gene, testis-specific basic protein (TSBP). We named the new locus BTL-II, for butyrophilin-like MHC class II associated. The six discernable exons of the BTL-II locus encode a small hydrophobic amino acid sequence (which may be a signal peptide), two immunoglobulin domains, a small 7 amino acid, heptad repeat-like exon, and a further two immunoglobulin domains. In mouse, an additional butyrophilin-like gene (NG10) is situated adjacent to BTL II. Expression studies of the BTL-II locus in mouse showed that it is expressed in a range of gut tissues. We demonstrate that like many other genes from the MHC, BTL-II is polymorphic in a selection of diverse HLA haplotypes. In the light of the newly discovered locus, we revisit and discuss the possible origin of the butyrophilin gene family. PMID- 10803853 TI - G7c, a novel gene in the mouse and human major histocompatibility complex class III region, possibly controlling lung tumor susceptibility. PMID- 10803854 TI - Molecular cloning and sequencing of cDNA encoding eight new rhesus macaque TCRBV gene families. PMID- 10803855 TI - Polymorphism of the mouse T-cell receptor AV20S1 gene. PMID- 10803856 TI - Implications of the UK prospective diabetes study: questions answered and issues remaining. AB - The UK Prospective Diabetes Study (UKPDS) provides the first conclusive proof for the importance of intensifying diabetes control in individuals with type 2 diabetes mellitus. However, reduction in cardiovascular disease risk with intensive therapy was modest and did not reach statistical significance. Metformin therapy in obese individuals with type 2 diabetes mellitus was associated with reduced cardiovascular death. These observations should be re evaluated to determine whether various therapeutic agents available for treatment of type 2 diabetes mellitus have different effects on cardiovascular complications of diabetes. The addition of alpha-glucosidase inhibitor, acarbose, improved glycaemic control irrespective of concomitant therapy for diabetes, although compliance with this agent was poor. The tight blood pressure control study embedded in UKPDS reaffirms the importance of lowering the blood pressure below 150/85 to reduce microvascular and macrovascular complications of diabetes. PMID- 10803857 TI - Differential pharmacokinetics of lithium in elderly patients. AB - The pharmacotherapeutic use of lithium in the elderly as acute and maintenance therapy in bipolar disorder and augmentation therapy for major depression is well documented. Differences in the response to lithium are explained, in part, by the effect of age-related physiological changes, comorbid conditions, and concurrent medications on the pharmacokinetics of lithium in the elderly. The pharmacokinetic profile of lithium has been studied for many years, primarily in younger adult populations. Lithium pharmacokinetics may be influenced by a number of factors including age. It was first noted several years ago that elderly individuals required lower doses of lithium to achieve serum concentrations similar to those observed in younger adults. This is due to the combination of a reduced volume of distribution and reduced renal clearance. The composition of the human body changes with aging producing an increase in body fat, a decrease in fat-free mass and a decrease in total body water. Lithium clearance decreases as the glomerular filtration rate decreases with increasing age. The effects of other medical conditions in the elderly on the pharmacokinetics of lithium are less well delineated. Reduced lithium clearance is expected in patients with hypertension, congestive heart failure or renal dysfunction. Larger lithium maintenance doses are required in obese compared with non-obese patients. The most clinically significant pharmacokinetic drug interactions associated with lithium involve drugs which are commonly used in the elderly. Thiazide diuretics, ACE inhibitors, and nonsteroidal anti-inflammatory drugs can increase serum lithium concentrations. The tolerability of lithium is lower in the elderly. Neurotoxicity clearly occurs in the elderly at concentrations considered 'therapeutic' in general adult populations. There are no placebo-controlled randomised trials of lithium in old age and recommendations for clinical use are based on extrapolations from pharmacokinetic studies, anecdotal reports from mixed age populations and clinical experience in old age psychiatry. Serum concentrations of lithium need to be markedly reduced in the elderly population and particularly so in the very old and frail elderly. PMID- 10803858 TI - Platelet glycoprotein IIb/IIIa receptor antagonists and their use in elderly patients. AB - With increasing age of the general population, cardiovascular diseases are becoming a greater health burden. Coronary artery disease remains a major cause of morbidity and mortality worldwide. Among the various pathophysiological processes, platelets play a pre-eminent role. With the identification of the glycoprotein (GP) IIb/IIIa receptor as the final common pathway for platelet aggregation, potent antiplatelet agents have been developed. These GP IIb/IIIa antagonists have been shown to be effective in improving outcomes among patients undergoing percutaneous coronary interventions and for the treatment of acute coronary syndromes. By pooling the results of several large-scale trials, these benefits have been found to extend to the elderly population. Among 7860 patients undergoing percutaneous coronary intervention, the occurrence of death or myocardial infarction at 30 days was reduced from 10.0 to 5.9% (odds ratio 0.56; 95% confidence level, 0.37 to 0.83) with abciximab compared with placebo, in those >70 years of age. Importantly, this benefit was achieved without an increase in major bleeding complications. Similarly favourable trends were also observed among elderly patients treated with tirofiban or eptifibatide for acute coronary syndromes. As such, GP IIb/IIIa antagonists are effective in preventing ischaemic complications and can be safely administered to elderly patients. PMID- 10803859 TI - St John's wort: a potential therapy for elderly depressed patients? AB - St John's wort (Hypericum perforatum) has a 2000-year history of use as a medicinal herb. Its modern application as a plant extract for treating depression has undergone scientific investigation over the last decade, and its effectiveness has been shown in studies comparing it with placebo and reference antidepressants. Our own work supports the contention that LI 160, the best documented St John's wort medication, is effective at a high dosage even in patients with severe depression. Since the new Berlin Aging Study revealed significant treatment deficiencies among elderly depressive patients, St John's wort extracts may be a useful alternative to benzodiazepines to avoid nontreatment of early depression. Because St John's wort preparations have better tolerability than tricyclic antidepressants, elderly people in particular, can benefit from their use. PMID- 10803860 TI - Donepezil: a review of its use in Alzheimer's disease. AB - Donepezil (E-2020) is a reversible, noncompetitive, piperidine-type cholinesterase inhibitor. It is selective for acetylcholinesterase rather than butyrylcholinesterase. Donepezil 5 and 10 mg/day significantly improved cognition and global clinical function compared with placebo in well designed short term trials (14 to 30 weeks) in 161 to 818 patients with mild to moderate Alzheimer's disease. Beneficial effects on cognition were observed from week 3 of treatment. Donepezil 10 mg/day significantly delayed the deterioration in activities of daily living (ADL) [by 55 weeks] compared with placebo in a retrospective analysis of 1 trial, and in the largest trial significantly improved patients' abilities to perform complex tasks. However, no significant improvement in function was observed with donepezil 5 mg/day in another trial. In the 2 trials of longest duration donepezil (5 and 10 mg) significantly delayed symptomatic progression of the disease. While there was no evidence for a positive effect of donepezil on patients' quality of life, there are no validated measures of this parameter specific to patients with Alzheimer's disease. Donepezil (5 and 10 mg) significantly reduced caregiver burden. Long term efficacy data suggest that improvements in cognition, global function or ADL are maintained for about 21 to 81 weeks with donepezil (10 mg/day in most patients). Donepezil is generally well tolerated with the majority of adverse events being mild and transient. Predictably, most events were cholinergic in nature and generally related to the gastrointestinal and nervous systems. The incidence of these events was significantly higher with donepezil 10 mg than with placebo in short term clinical trials; however, this may have been due to the 7-day dose increase schedule used in these studies and can be minimised by increasing the dose after a longer (6-week) period. The incidence of serious adverse events was generally similar between donepezil 5 and 10 mg (4 to 10%) and placebo (5 to 9%) in short term trials. 26% of patients receiving donepezil (5 and 10 mg) reported serious events over a 98-week period in a long term trial. Importantly, there was no evidence of hepatotoxicity with this drug. Conclusions. Donepezil (5 and 10 mg) is an agent with a simple once-daily dosage schedule which improves cognition and global clinical function in the short (up to 24 weeks) and long term (up to about 1 year) in patients with mild to moderate Alzheimer's disease. Improvements in ADL were also observed with donepezil 10 mg/day. Adverse events associated with donepezil are mainly cholinergic. Donepezil has been extensively studied and should be considered as a first-line treatment in patients with mild to moderate Alzheimer's disease. PMID- 10803862 TI - A concise genetic and clinical guide to multiple endocrine neoplasias and related syndromes. AB - Several familial neoplastic syndromes are associated with endocrine gland oncogenesis. The main ones are: multiple endocrine neoplasia type 1 (MEN 1), which affects primarily the pituitary, pancreas, and parathyroid glands; MEN 2A and MEN 2B, which involve mainly the thyroid and parathyroid glands and the adrenal medulla; familial medullary thyroid carcinoma (FMTC), which affects only the thyroid gland; and, finally, Carney complex, which affects the adrenal cortex, pituitary, thyroid gland, and the gonads. Carney complex is also associated with pigmentation abnormalities and myxoid and other neoplasms of mesenchymal origin. Thus, this syndrome also belongs to another group of genetic disorders, those associated with pigmentation defects and multiple tumors, including tumors of the endocrine glands. Peutz-Jeghers syndrome and Cowden disease are just two of these disorders that have recently been elucidated at the molecular level. von Hippel-Lindau disease is another condition that affects the pancreas and adrenal medulla and its gene is also known. The inheritance of the MENs, Carney complex, and related syndromes is autosomal dominant. Clinical recognition of these syndromes at a young age improves clinical outcome and prognosis of the various tumors and decreases associated morbidity and mortality. This review considers a wider, more inclusive view of the MEN syndromes, summarizes their clinical features and presents the newest information on their molecular elucidation. PMID- 10803863 TI - Detection and management of diabetic glomerulopathy in children and adolescents with insulin-dependent diabetes mellitus: need for improved knowledge and better care. AB - Current modalities of detection and management of incipient diabetic nephropathy in childhood and adolescence are summarized and the open questions outlined. In particular, the predictive value of microalbuminuria for later nephropathy in adolescents is examined. Microalbuminuria is not as strong a predictor as suggested in earlier studies - recent studies show up to 50% of microalbuminuria can revert to normal. We advocate that the association of well-established risk markers and promoters of renal injury, including degree and tracking of albuminuria, glycemic control, blood pressure changes, incipient retinopathy and genetic background, allow more precise assessment of the individual risk of developing nephropathy and the decision to start pharmacological intervention. The major impact of strict glycemic control to prevent the development and progression of diabetic nephropathy is emphasized, as well as the need for a multidisciplinary team to optimize the care of pediatric diabetic patients. We discuss other therapeutic options, i.e. angiotensin-converting enzyme inhibitors (ACE-Is), moderate dietary protein intake, and other drugs. ACE-Is may provide a second line intervention in a well selected, high-risk subgroup of microalbuminuric diabetic adolescents. PMID- 10803861 TI - Losartan: a review of its use, with special focus on elderly patients. AB - Losartan is an orally active, nonpeptide, selective angiotensin subtype 1 (AT1) receptor antagonist. It provides a more specific and complete blockade of the actions of angiotensin II than renin or ACE inhibitors. Short term (up to 12 weeks' duration) clinical trials have shown losartan to be as effective at lowering blood pressure (BP) [causes a decrease in BP < or = 26/20 mm Hg] in elderly patients with hypertension as recommended dosages of captopril, atenolol, enalapril, felodipine and nifedipine. In patients with isolated systolic hypertension (ISH) the efficacy of losartan was similar to that of atenolol. The addition of hydrochlorothiazide to losartan therapy provides greater antihypertensive efficacy, equivalent to that seen with captopril plus hydrochlorothiazide. Preliminary evidence also indicates that losartan therapy contributes to the regression of left ventricular hypertrophy associated with chronic hypertension. Exercise capacity is increased by losartan in patients with either asymptomatic or symptomatic heart failure. Results from the Losartan Heart Failure Survival or ELITE II (Evaluation of Losartan in the Elderly II) study indicate that there was no statistically significant difference between losartan and captopril in reducing overall deaths or in reducing sudden cardiac death and/or resuscitated cardiac arrest in patients with heart failure. Other than ELITE II, little conclusive long term mortality and morbidity data exist for losartan. Additional long term trials to evaluate the survival benefits of losartan in elderly patients with hypertension, renal disease or after an acute myocardial infarction are currently in progress. In elderly patients with hypertension, the incidence of treatment-related adverse events associated with once daily losartan (alone or in combination with hydrochlorothiazide) [19 to 27%] was similar to felodipine (23%) and nifedipine (21%), however, losartan tended to be better tolerated than captopril (11 vs 16%). Losartan was also better tolerated than atenolol in patients with ISH (10.4 vs 23%). In patients with heart failure the renal tolerability of losartan was similar to that of captopril, but losartan was associated with a lower withdrawal rate because of adverse events. No dosage adjustment is required in elderly or in patients with mild to moderate renal dysfunction, and the risk of first-dose hypotension is low. CONCLUSIONS: comparative data have shown losartan to be as effective as other antihypertensive agents in the treatment of elderly patients with hypertension. Treatment with losartan is therefore an option for first-line therapy in all patients with hypertension, particularly those who are not well managed with or who are intolerant of their current therapy. Morbidity and mortality data from the Losartan Heart Failure Survival (ELITE II) study show that losartan has potential in the treatment of heart failure. PMID- 10803865 TI - Expression of the aromatase gene in the human prepubertal testis. AB - It has been proposed that estrogens might play a negative feedback role in the local regulation of androgen biosynthesis in the testis. Although aromatase has been reported to be present in human adult Leydig cells, CYP19 gene expression in the human prepubertal testis has not been studied. Human prepubertal testicular tissue was obtained from 12 testes collected at necropsy. Ages ranged from 0.07 to 7 years, but 7 of the 12 subjects were younger than 3 months old. Tissue mRNA was subjected to RT-PCR analysis by two methods. Cytochrome P450arom mRNA was detected by non-radioactive RT-PCR in five of the 12 prepubertal testes collected from 0.05-7 year-old subjects, and in one testis collected from a 15 year-old pubertal control. Four of these five prepubertal samples belonged to the youngest infant group. Using a more sensitive, radioactive RT-PCR, aromatase mRNA was detected in all prepubertal testes. This study shows that the CYP19 gene is expressed in the prepubertal human testis including the period of early postnatal activation. It is possible that estrogens may have a role in prepubertal males during this period. PMID- 10803864 TI - Low serum TNF-alpha levels in subjects at risk for type 1 diabetes. AB - OBJECTIVE: The aim of our study was to determine whether serum TNF-alpha levels in individuals at risk of developing type 1 diabetes, such as first-degree relatives of diabetic patients and children with incidental hyperglycemia, underwent alterations, and also to establish whether these levels might be used to identify individuals prior to insulin dependence. RESEARCH DESIGN AND METHOD: We studied 71 healthy first-degree relatives (FDR) of type 1 diabetic patients and 11 children with incidental hyperglycemia. We looked for immunogenetic (HLA class II serologic alleles and HLA-DQ alpha/beta genomic polymorphisms), immunologic (islet-cell and insulin autoantibodies) and metabolic (FPIR to IVGTT) markers of type 1 diabetic risk. Serum concentrations of TNF-alpha were quantified using IRMA. RESULTS: We found significantly lower serum TNF-alpha levels in FDR of type 1 diabetic patients (median: 54.3 pg/ml) (p=0.01) and in children with incidental hyperglycemia (median: 10.83 pg/ml) (p<0.0001) compared to controls (median: 76.56 pg/ml). No significant difference was observed between subjects with or without immunogenetic, immunologic and metabolic markers of type 1 diabetic risk. A negative correlation was found between serum TNF-alpha and HbA1c levels (r=-0.27, p=0.023). Two children with incidental hyperglycemia, whose TNF-alpha levels were very low, developed type 1 diabetes 6 and 8 months after this study. CONCLUSION: Our results are compatible with an impaired immune system in the prediabetic period and suggest that serum TNF-alpha concentrations may be considered as an immunological marker useful to identify subjects at risk of developing type 1 diabetes. PMID- 10803866 TI - Polymorphism in the transmembrane region of the MICA gene and type 1 diabetes. AB - Although MHC class II genes have a stronger association with type 1 diabetes than MHC class I genes, studies have shown that MHC class I molecules play an independent role in the etiology of type 1 diabetes, and the existence of susceptibility genes within a segment of MHC between the HLA-B and TNF genes has been predicted, where MHC class I chain-related gene A (MICA) resides. MICA has a triplet repeat polymorphism in the transmembrane region consisting of five alleles. We analyzed this polymorphism in 162 unrelated children (82 boys) with type 1 diabetes (age at diagnosis 7.01 +/- 3.76 yr) and 154 randomly selected unrelated children (87 boys), age 2.81 +/- 2.12 yr. Phenotype frequency of allele A9 in children with type 1 diabetes was significantly higher than in controls (RR = 2.42, 95% CI = 1.52-3.85, p = 0.000162, pc = 0.00081). Gene frequency of allele A9 was also significantly higher in children with type 1 diabetes when compared with control children (RR = 2.73, 95% CI = 1.85-4.03, p = 2.62 x 10(-7), pc = 1.31 x 10(-6)). This study demonstrates that MICA allele A9 confers risk of type 1 diabetes. PMID- 10803867 TI - Ethnic differences in androgens, IGF-I and body fat in healthy prepubertal girls. AB - OBJECTIVE: To examine ethnic differences in adrenal androgen production, IGF-I, and IGFBP-1 and -3 in relation to bone age, insulin, and body composition in healthy prepubertal girls. METHODS: Serum levels of DHEA-S, androstenedione, IGF I, and IGFBP-1 and -3 were examined in relation to bone age, insulin, and body composition (determined by dual-energy X-ray absorptiometry) in 47 (19 Caucasian, 9 African-American, 19 Mexican-American) healthy prepubertal girls aged 7.5-9.0 years. RESULTS: Age, weight, height, bone age, androstenedione, insulin, glucose:insulin ratios, and IGFBP-3 levels were not statistically different among groups. Mexican-American girls had higher % body fat than African-Americans or Caucasians (P < 0.001). DHEA-S levels in African-Americans were twofold higher than in Caucasians (P = 0.024), although their % body fat was not significantly different (16.1% and 19.4%, respectively; P = 0.138). DHEA-S levels in Mexican American girls were intermediate. Bone age and weight were significant covariates for DHEA-S levels. Plasma IGF-I levels were also higher in African-American than in Caucasian or Mexican-American girls (P = 0.009). Covariance analysis showed that IGF-I levels were influenced mainly by ethnicity (P = 0.009) and were independent of bone age. Despite similar insulin levels among groups, IGFBP-1 levels were higher in Caucasians than in Mexican-Americans or African-Americans (P < 0.001). CONCLUSIONS: In healthy prepubertal girls, DHEA-S concentrations are higher in African-Americans than in Caucasians or Mexican-Americans, even before any clinical evidence of adrenarche. Furthermore, IGF-I concentrations are higher in African-American girls than in Caucasian or Mexican-American girls which may contribute to the higher DHEA-S levels observed. Conversely, higher DHEA-S and IGF-I levels in African-American girls may be indicative of an influence not only of gonadal but also of adrenal androgens on the GH/IGF-I axis. PMID- 10803868 TI - The relationship between different subcutaneous adipose tissue layers, fat mass and leptin in obese children and adolescents. AB - We studied the relationships of subcutaneous adipose tissue layers (SAT-layers), body fat mass (FM) and waist-to-hip ratio (WHR) with leptin in obese children and adolescents. Twenty-nine obese children and adolescents (12 boys: age: 11.3 +/- 3.7 yr; body mass index [BMI]: 28.5 +/- 4) and 17 girls (age: 12.2 +/- 2.2 yr; BMI: 29.8 +/- 4.7) (mean +/- SD) were studied. FM was estimated by bioelectrical impedance. SAT-layers were determined at 15 different body sites from 1-neck to 15-calf by the Lipometer optical device. Leptin and insulin were determined by RIA. Maturity was associated with a greater thickness of certain SAT-layers from the upper body and with a lower thickness of SAT-layers from the abdominal region and lower extremities. Significant correlations were found for all estimates of adiposity and leptin (all p<0.001). Waist and hip circumferences were not correlated to leptin after adjustment for FM. SAT-layers from the upper body were significantly and positively correlated to leptin. Multiple regression analysis revealed FM as a main contributor to the variation in leptin (R2=0.53, p<0.0001). FM together with SAT-layers 5-front chest and 13-rear thigh explained 72% of the variation in leptin (p<0.0001). In a body fat distribution model, hip circumference together with SAT-layers 4-upper back and 2-triceps explained 75% of the variation in leptin (p< 0.0001). The results suggest that SAT-layers and their topography are main determinants for leptin in obese children and adolescents. Maturity in obese children is associated with higher values of upper body SAT-layers and lower values of abdominal and lower extremities SAT-layers. Whether leptin is under the control of certain subcutaneous adipose tissue depots from the upper body remains to be elucidated by longitudinal studies. PMID- 10803869 TI - No evidence for leptin as an independent associate of blood pressure in childhood and juvenile obesity. AB - We studied whether leptin is an independent associate of blood pressure in obese children and adolescence. 102 obese children (48 girls, age: 11.6 +/- 2.22 yr; body mass index [BMI]: 27.45 +/- 4.4; blood pressure: 122.5 +/- 11.1/64.7 +/- 10.6 mm Hg and 54 boys, age: 11.5 +/- 2.4 yr; BMI: 27.6 +/- 4.4; blood pressure: 122.5 +/- 13.2/60.9 +/- 8.1 mm Hg [mean +/- SD]) were investigated. Serum leptin and insulin were measured by RIA; glucose was determined enzymatically. Fat mass (FM) was calculated by bioelectrical impedance. Leptin was higher in girls than in boys (p=0.018) but no significant gender differences were found with respect to indices of adiposity and systolic blood pressure (SBP). Children were divided into three groups, according to pubertal stage (Group 1: prepubertal, 32 boys/13 girls; Group 2: pubertal, 17 boys/25 girls; Group 3: late/postpubertal, 5 boys/10 girls). SBP and DBP correlated with body weight in the whole group (r=0.49, p<0.0001, and r=0.27, p=0.004). In Group 1, BMI showed the highest correlation to SBP; in Group 3 no indices of adiposity were related to SBP. In no case was leptin significantly associated with SBP after adjustment for adiposity. In Group 2, glucose was significantly associated with SBP after adjustment for body weight. In Group 3, however, no correlations were found between SBP, DBP and metabolic characteristics, perhaps due to small sample size. Stepwise multiple regression revealed that body weight and glucose contributed to the variation in SBP in the whole group (R2=0.31, p<0.0001). Insulin accounted for almost 8% of the variation in DBP (R2=0.08, p=0.0034). Body weight contributed significantly to SBP in boys (R2=0.39, p<0.0001) and girls (R2=0.24, p< 0.001). The results imply that body weight contributes independently to the variation in blood pressure. Glucose and insulin contribute to mean blood pressure to some extent, but our data do not support the assumption that leptin per se serves as an independent predictor of blood pressure in obese children and adolescents. PMID- 10803870 TI - Coxsackie infection and births of future diabetic children: year, seasonality and secularity. AB - BACKGROUND: Environmental factors including Coxsackie virus infections are considered as one of the etiopathogenetic mechanisms in childhood insulin dependent diabetes mellitus (IDDM). HYPOTHESIS: This relationship should be reflected not only in similar seasonal and secular periodicities of Coxsackie epidemics and IDDM incidence but also in the cycling of births of future diabetic children. SUBJECTS: Seven hundred and seven healthy Slovak children randomly selected in 1985-98 were investigated for the presence of actual Coxsackie infection. The months of birth in 1970-93 of 1779 Slovak children with later IDDM were registered. AntiCoxsackie positivity was determined in 1978-91 at IDDM onset in 336 children. METHODS: AntiCoxsackie positivity was assessed by IgM antibodies and virus neutralization test. Time series data were processed by cosinor regression enabling the definition of statistical significance of trends and of cycles with tested period lengths. RESULTS: Marked autumnal peaks were found for Coxsackie infections. For births of future diabetics, both autumn and spring peaks were encountered. Moreover, seven-year cycling displayed peaks in 1990 and 1997 for Coxsackie and 1976, 1983 and 1990 for IDDM births. CONCLUSIONS: Connection between Coxsackie viral infections and births of future diabetic children appears plausible also in Slovakia. PMID- 10803871 TI - Seasonal variation of HbA1c in intensive treatment of children with type 1 diabetes. AB - OBJECTIVE: The aim of this study was to measure whether there is a seasonal variation in glycosylated haemoglobin concentrations and insulin dose used in the intensive treatment of children with type 1 diabetes, and whether such variation is related to severe hypoglycaemia. PATIENTS: A geographic population of 114 intensively treated type 1 diabetic patients < 19 years of age, mean 12.7 (SD 4.3) years, with diabetes onset before 1995, were studied in a cohort 1995-96. METHODS: HbA1c, insulin doses and severe hypoglycaemia were registered at regular visits scheduled quarterly, but not standardised in time. Seasonal mean values were calculated for HbA1c and insulin dose. RESULTS: Lower HbA1c was seen in spring and summer, and higher in autumn and winter (p=0.023). Patients reporting severe hypoglycaemia had a seasonal variation in HbA1c (p=0.019) and a tendency to seasonal variation in insulin dose, while patients not reporting severe hypoglycaemia did not vary in HbA1c or insulin dose. CONCLUSIONS: Self-control and adjustment of insulin doses to seasonal change need to be improved also in intensively treated children, with regard to the risk for worsened metabolic control after the summer and increased severe hypoglycaemia in spring and early summer. The findings have important implications for design of short-term studies of metabolic control. PMID- 10803872 TI - Linear growth and weight gain in diabetic children--a cross-sectional and longitudinal evaluation. AB - OBJECTIVE: To examine linear growth and weight gain in diabetic children and to assess the influence of the age at onset of diabetes on growth. SUBJECTS AND METHODS: A retrospective longitudinal and cross-sectional analysis of the growth data of 61 children attending the diabetic clinic for the whole year of 1998 was completed. RESULTS: The children were of average height and weight at onset with mean (SD) Height SDS = -0.095 (0.96) and mean (SD) body mass index (BMI) SDS = 0.58 (1.15). But amongst the subgroups, boys with onset before five years were found to be significantly taller (Height SDS = 0.39 (0.75), P<0.05) and heavier (BMI SDS = 1.28 (1.05), P<0.05). There was no significant change from onset to time at analysis either within individual subgroups or as a whole. Girls showed a gain in mean (SD) BMI SDS from 0.41 (1.14) to 1.03 (1.25), which however did not reach statistical significance (P=0.08). No similar tendency was observed in boys. There was no significant change in Height SDS for the 12 children who reached final height. Longitudinal follow up of growth data agreed with the observations from cross-sectional data. CONCLUSION: Prepubertal linear growth was not affected even in those children diagnosed early in childhood. The 12 children who reached final height did not show any impairment of growth. Increased growth at onset was observed in the subgroup of boys with onset of diabetes before 5 years. The tendency towards excessive weight gain observed in diabetic girls needs further evaluation. PMID- 10803873 TI - The effect of 3 years of recombinant growth hormone therapy on glucose metabolism in short Chinese children with beta-thalassemia major. AB - Growth retardation and diabetes mellitus are common in children and adolescents with beta-thalassemia major despite hypertransfusion regimen and iron chelation therapy. The purpose of this study was to investigate the effects of growth hormone (GH) treatment on glucose metabolism in children with beta-thalassemia major. GH therapy for 3 years improved the height SD scores of eight short prepubertal Chinese children with beta-thalassemia major from -2.15 +/- 0.90 to 1.14 +/- 0.78 (paired t-test, p = 0.01) without excessive advancement in bone age (ABA/CA = 0.95 +/- 0.27). There was no deleterious effect on glucose metabolism with no change in fasting blood sugar, serum fructosamine, fasting and stimulated insulin to intravenous glucose infusion (sum of 1+3 min insulin, In 1+3'; incremental insulin 0-10 min area above fasting concentrations, deltaInAUC0-10'; ratio of incremental 0-10 min insulin area above fasting concentrations over glucose area above fasting concentrations, delta0-10'AUCIn/G; ratio of incremental 0-10 min insulin over peak glucose above basal 0-10 min, delta0 10'InAUC/deltaGPeak), and glucose disappearance coefficient (Kg). Short term GH therapy improves the height of children with beta-thalassemia major but the effect of treatment on final height still needs to be determined. PMID- 10803874 TI - Helicobacter pylori infection in children with insulin dependent diabetes mellitus. AB - To assess the seroprevalence of Helicobacter pylori (HP) in children with insulin dependent diabetes mellitus, a serological test for Helicobacter pylori (anti-HP IgG with ELISA) was performed in 88 diabetic and 42 healthy control children. Anti-HP IgG was positive in 49/88 (55.6%) of diabetics and 13/42 (30.9%) of controls (p<0.01). Diabetic children were divided into two groups according to HP status: HP(+) and HP(-). The two groups were compared for age, gender, duration of diabetes, diabetic control (HbA1c), SDS for height and gastric emptying time. Seroprevalence of HP was higher in IDDM patients than in healthy controls. Duration of diabetes was the only factor which correlated significantly with HP status. HP status was not related to gastric emptying time. PMID- 10803875 TI - Association of Turner's syndrome and Swyer's syndrome in the same family. AB - We report two phenotypically and genetically different diseases in the same family. One patient presented with Turner phenotype as a result of chromosomal mosaicism 45,X/46,X, inv(X)(q21;q24) (30%/70%). Her father's sister showed 46,XY female gonadal dysgenesis (Swyer's syndrome) as a result of a point mutation in the SRY gene on her Y chromosome. DNA sequencing revealed a G-->C transversion (nucleotide position 693) resulting in a change from glycine95 to arginine (G95R). Here we report for the first time an association of Turner's syndrome and Swyer's syndrome in the same family. PMID- 10803876 TI - A patient with congenital hypertrichosis, gum hyperplasia and macromastia. AB - A syndrome of congenital hypertrichosis, hirsutism, gum hyperplasia and macromastia is described. The patient was demonstrated to have mild hyperinsulinemia with normal oral glucose tolerance test. This is the second such patient reported in the literature. PMID- 10803877 TI - Langerhans' cell histiocytosis: report of an atypical case. AB - A patient is described with Langerhans' cell histiocytosis and polyneuropathy diagnosed 12 years after the development of diabetes insipidus after head trauma. PMID- 10803878 TI - Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections. AB - Oral administration of specific antibodies is an attractive approach to establish protective immunity against gastrointestinal pathogens in humans and animals. The increasing number of antibiotic-resistant bacteria emphasize the need to find alternatives to antibiotics. Immunotherapy can also be used against pathogens that are difficult to treat with traditional antibiotics. Laying hens are very good producers of specific antibodies. After immunization, the specific antibodies are transported to the egg yolk from which the antibodies then can be purified. A laying hen produces more than 20 g of yolk antibodies (IgY) per year. These antibodies also have biochemical properties that make them attractive for peroral immunotherapy: They neither activate mammalian complement nor interact with mammalian Fc receptors that could mediate inflammatory response in the gastrointestinal tract. Eggs are also normal dietary components and thus there is practically no risk of toxic side effects of IgY. Yolk antibodies have been shown in several studies to prevent bacterial and viral infections. PMID- 10803879 TI - Multi-envelope HIV vaccine safety and immunogenicity in small animals and chimpanzees. AB - A significant obstacle to HIV vaccine development lies in the remarkable diversity of envelope proteins, the major targets of neutralizing antibody. That envelope diversity must be targeted is demonstrated by results from nonhuman primate studies in which single-envelope vaccines have protected against homologous, but rarely against heterologous virus challenges. Similarly, in clinical trials, single-envelope vaccines have failed to prevent break-through infections when challenge viruses were inevitably mismatched with the vaccine. To protect humans from infection by any isolate of HIV, we have prepared vaccine cocktails combining multiple envelopes from distinct viral isolates. We have tested several vehicles for vaccine delivery in small animals and have shown that successive immunizations with envelope, presented first as a DNA recombinant, then as a vaccinia virus (VV) recombinant, and finally as purified protein elicited strong neutralizing antibody responses. We have also tested the VV recombinant vaccine in chimpanzees. Pairs of animals received either single- or multi-envelope VV recombinant vaccines administered by the subcutaneous route. Results showed that the multi-envelope vaccine was safe, immunogenic, and superior to the single-envelope vaccine in eliciting HIV-specific antibody measurable in a standard clinical, immune assay. The promise of this system has led to the initiation of clinical trials, with which the hypothesis that cocktail vaccines will prevent human HIV infections may ultimately be tested. PMID- 10803880 TI - CD27/CD70 interactions regulate T dependent B cell differentiation. AB - CD27 is a tumor necrosis factor (TNF) receptor family member whose expression is limited to cells of the lymphoid lineage. Constitutively expressed on T lymphocytes, it is a costimulatory molecule for a regulatory subset. Induced on B lymphocytes after antigenic challenge, it is a marker of memory cells. CD70, CD27 ligand, is a TNF related trans-membrane protein induced upon activation on T and B cells. In complement of ligation of CD40, another TNF receptor family member expressed by B cells, CD27/CD70 interaction plays a key role in T dependent B cell responses and is responsible for plasma cell differentiation. B lymphocyte responses appear thus controlled by different T cell subsets expressing CD154 (CD40 ligand), CD27, or CD70 (CD27 ligand). PMID- 10803881 TI - Cell proliferation and apoptosis in the immune system in the elderly. AB - Loss of the cell proliferative capability and involution of tissues and organs are among the most important phenomena that characterize the aging process. Some of the aged-linked immune dysfunctions could be partly due to a dysregulation of apoptotic processes and to a lower responsiveness of aged lymphoid cells to activation and proliferation signals. The main changes in proliferative activity and cell death during aging and their impact on the process of immunosenescence are discussed. In fact, a very important function that has been suggested to deteriorate with age and to play a major role in the aging process is the capability of cells from aged subjects to respond to mitogenic stimuli and, consequently, to undergo cell proliferation. However, the cellular activation processes are very complex and the proliferative responses can follow different interconnected signal transduction pathways, and only some of them appear to be modified during age. Moreover, cell growth, immunosenescence, and longevity are strictly interconnected and deeply related to programmed cell death or apoptosis. The cellular equilibrium between cell survival and proliferation, on the one hand, and programmed cell death, on the other hand, seems to be unbalanced with advancing age, although in each type of immune cell it could be differentially modulated, resulting in a variety of clinicopathological consequences. Thus, cell proliferation and cell death are two physiologically active phenomena closely linked and regulated and a failure of these mechanisms determines profound dysregulations of cell homeostasis with major consequences in immune functioning and the onset of autoimmune diseases and cancer, whose incidence appears to be increased in the elderly. PMID- 10803882 TI - Ly49 gene expression in different inbred mouse strains. AB - Mouse natural killer cells express receptors for class I MHC in the form of the Ly49 family of proteins. The Ly49 family contains at least 13 expressed members (A, B, C, D, E, F, G, H, I, J, L, O, and P) and is further subdivided into activating and inhibitory subfamilies based on intracellular and transmembrane characteristics. The level of sequence identity between different members varies dramatically. However, comparison of the extracellular domain has revealed that several of the Ly49 molecules also form "pairs," where one member is activating and the other is inhibitory. Until recently, most Ly49 molecules described have come from the C57B1/6 strain of inbred mice. Using molecular cloning and immunochemical analysis we have found that different mouse strains express novel Ly49 molecules. Comparison of the allelic forms of some Ly49 molecules has shown that the dividing line between different genes and different alleles is blurred. PMID- 10803883 TI - Mss51p, a putative translational activator of cytochrome c oxidase subunit-1 (COX1) mRNA, is required for synthesis of Cox1p in Saccharomyces cerevisiae. AB - Mutants of Saccharomyces cerevisiae that lack a functional MSS51 gene are respiratory deficient due to the absence of cytochrome c oxidase subunit 1 (Cox1p). It has been previously suggested, but not formally proven, that Mss51p is required for translational activation of COX1 mRNA, rather than being involved in a subsequent step in the synthesis of Cox1p or its assembly into cytochrome c oxidase. Pulse-chase labelling experiments now show that the absence of detectable levels of Cox1p in mss51-null strains is indeed due to the lack of synthesis of Cox1p, and is not caused by reduced stability of the protein. To gain more insight into the exact function of Mss51p, we determined the subcellular localization of the protein. We were able to show that an epitope tagged version of Mss51p (Mss51HA) complements the mutation and can be localized in mitochondria, where it is firmly associated with the mitochondrial inner membrane. In addition, we characterized the previously identified mutant allele mss51-3. Sequence analysis revealed the presence of a short open reading frame upstream of MSS51 resulting from the creation of an extra ATG start-codon. PMID- 10803884 TI - Characterization of novel rad6/ubc2 ubiquitin-conjugating enzyme mutants in yeast. AB - Null mutations in the RAD6/UBC2 gene encoding an E2 ubiquitin-conjugating enzyme cause deficiencies in DNA repair, N-end-rule protein degradation, sporulation and telomeric silencing, and alter the preferred integration positions for Ty1 retrotransposons. Here we selected for mutants of RAD6 that cause a release of telomeric silencing. Some alleles retained nearly wild-type ability for sporulation, DNA repair and the degradation of proteins. Alteration in Ty1 integration-site bias accompanied some of these alleles. The possibility that some mutations specifically affect binding of an unknown protein that works with Rad6 in its silencing role, but is not required for DNA repair or N-end-rule activity, is discussed in terms of the Rad6 crystal structure. PMID- 10803885 TI - Antimycin A resistance in a mutant Leishmania tarentolae strain is correlated to a point mutation in the mitochondrial apocytochrome b gene. AB - In this paper we report the first case of antimycin A resistance in a protozoan parasite that is attributable to a mutation in the mitochondrial apocytochrome b (CYb) gene. We selected for, and isolated, a mutant Leishmania tarentolae strain that is resistant to antimycin A. This resistance was evident at the levels of the in vitro growth and enzymatic activity of the cytochrome bc1 complex. Molecular characterisation of the mutant revealed a Ser35Ile mutation in the expected region of the CYb gene. In kinetoplastids, CYb and other structural genes of the mitochondrial genome are located on the maxicircle component of the mitochondrial DNA, which is present in 20-50 copies. Primer-extension analysis confirmed the presence of the mutation at the mRNA level. The phenotypic manifestation of the mutation implies that the CYb mRNA is edited and translated within the mitochondrion. Thus, this finding provides direct evidence that edited RNAs are translated in kinetoplastid mitochondria. Furthermore, a defined mutation conferring drug resistance to a mitochondrial gene product can be exploited for the development of mitochondrial transfection systems for trypanosomatids. PMID- 10803886 TI - A circular mitochondrial plasmid incites hypovirulence in some strains of Cryphonectria parasitica. AB - In the chestnut-blight fungus Cryphonectria parasitica, a plasmid, pCRY1, occurs in the mitochondria of several strains isolated at various locations in the northeastern United States and Canada. The monomer of this plasmid is a 4.2-kb circular double-stranded DNA that has no detectable sequence homology with the 160-kb mitochondrial DNA of Ep155, a standard virulent laboratory strain of C. parasitica. The circular nature and oligomeric characteristics of the plasmid were deduced from the heterogeneous size of plasmid DNA molecules as detected by one- and two-dimensional gel-electrophoresis, the nature and alignment of restriction fragments, and the lack of detectable termini in the nucleotide sequence. The cytoplasmic location of the plasmid was deduced from its co purification with mitochondria, uniparental (maternal) transmission in sexual crosses, dissociation from the nuclei of the donor strain during its horizontal transfer between vegetatively compatible strains through hyphal anastomoses, and mitochondrial codon usage (UGA = Try). The pCRY1 plasmid contains a long open reading frame that is transcribed and potentially encodes a unique 1214 amino acid, B-family DNA polymerase similar to those encoded by the LaBelle and Fiji circular mitochondrial plasmids of Neurospora. In this subgroup of proteins, the DTD motif characteristic of B-family DNA polymerases is replaced by TTD. Amino acid motifs related to those that are characteristic of the 3'-->5' exonuclease domains of B-family DNA polymerases have been located in the amino-terminal portion of the proteins. A comparison of isogenic plasmid-free and plasmid containing cultures indicates that pCRY1 is an infectious agent that effects a reduction in the pathogenicity of some, but not all, strains of C. parasitica. PMID- 10803887 TI - Dynamics of asexual transmission of a mitochondrial plasmid in Cryphonectria parasitica. AB - In the chestnut blight fungus Cryphonectria parasitica, as in most fungi, little is known about the efficiency of the asexual transmission of optional mitochondrial plasmids, vertically through conidia, and horizontally through hyphal anastomoses. In this paper, we show that pCRY1, a circular mitochondrial plasmid, is transmitted vertically with 100%-efficiency through conidia. Moreover, the plasmid is transmitted horizontally through hyphal contact from donor strains to vegetatively compatible and most incompatible strains. An allelic difference between the donor and recipient strain, at only one of the five nuclear incompatibility genes that were tested strongly inhibited, but did not absolutely prevent, the transfer of pCRY1 through hyphal fusions. In contrast, allelic differences in any one or several of the other four heterokaryon-compatibility loci suppressed the transmission of the plasmid only partially or not at all. The plasmid was also transmitted among incompatible strains by protoplast fusion without the concomitant transfer of mitochondrial DNA (mtDNA). A comparison of plasmid-bearing with plasmid-free isogenic strains revealed that pCRY1 significantly diminishes the pathogenic potency of some strains of the fungus, but does not affect the virulence of others. Collectively, the observations indicate that the introduction of deleterious mitochondrial genetic elements into natural populations may be a means for managing fungal pathogens. PMID- 10803888 TI - Copper-dependence of mitochondrial DNA rearrangements in Podospora anserina. AB - Rearrangements of the mitochondrial DNA (mtDNA) are a hallmark of senescence in wild-type strains of the ascomycete P. anserina. These rearrangements include the systematic amplification of the first intron (p1-intron) of the cytochrome oxidase subunit-I gene (CoI) as a circular DNA molecule (p1DNA). In addition, deletions and amplifications of other regions of the mtDNA occur. The molecular basis of the underlying processes is not understood in detail. A comparative analysis of the wild-type strain and of the long-lived mutant grisea, affected in the uptake of copper, revealed that mtDNA instabilities are dependent on the availability of cellular copper. In the mutant, the first steps in the corresponding pathway, including the transcription of the CoI gene, the splicing of the p1-intron and the transposition of this mobile element, are not impaired. In contrast, recombination processes between short direct repeats, as well as rearrangements between two tandem intron copies leading to the formation of p1DNA, appear to be affected. Additional copper in the growth medium rescues this molecular phenotype. We suggest that copper is a cofactor of a component of the molecular machinery leading to the characteristic age-related mtDNA rearrangements. PMID- 10803889 TI - Alternative sources of natural rubber. AB - Rubber (cis-1,4-polyisoprene) is one of the most important polymers naturally produced by plants because it is a strategic raw material used in more than 40,000 products, including more than 400 medical devices. The sole commercial source, at present, is natural rubber harvested from the Brazilian rubber tree, Hevea brasiliensis. Primarily due to its molecular structure and high molecular weight (> 1 million daltons) this rubber has high performance properties that cannot easily be mimicked by artificially produced polymers, such as those derived from, e.g., bacterial poly-hydroxyalkanoates (PHAs). These high performance properties include resilience, elasticity, abrasion resistance, efficient heat dispersion (minimizing heat build-up under friction), and impact resistance. Medical rubber gloves need to fit well, be break-resistant, allow the wearer to retain fine tactile sensation, and provide an effective barrier against pathogens. The sum of all these characteristics cannot yet be achieved using synthetic gloves. The lack of biodiversity in natural rubber production renders continuity of supply insecure, because of the risk of crop failure, diminishing acreage, and other disadvantages outlined below. A search for alternative sources of natural rubber production has already resulted in a large number of interesting plants and prospects for immediate industrial exploitation of guayule (Parthenium argentatum) as a source of high quality latex. Metabolic engineering will permit the production of new crops designed to accumulate new types of valued isoprenoid metabolites, such as rubber and carotenoids, and new combinations extractable from the same crop. Currently, experiments are underway to genetically improve guayule rubber production strains in both quantitative and qualitative respects. Since the choice for gene activities to be introduced or changed is under debate, we have set up a complementary approach to guayule with yeast species, which may more quickly show the applicability and relevance of genes selected. Although economic considerations may prevent commercial exploitation of new rubber-producing microorganisms, transgenic yeasts and bacteria may yield intermediate or alternative (poly-)isoprenes suitable for specific applications. PMID- 10803890 TI - Perspectives in the biological function and the technological application of polygalacturonases. AB - Polygalacturonases (PG) have evolved in the past years from a pectinase "simply" being used for food processing to an important parameter in plant-fungal interaction. PG-inhibiting proteins (PGIP) that are synthesised in plants as a specific response to PGs of pathogenic fungi, have become a focus as a possible target in resistance breeding, and PGIPs are also a concern as an inhibiting factor in food processing. Plant PGs have been identified as a major factor in fruit ripening, and PG-deficient transgenic plants have been bred. Mainly fungal PGs are used in industrial processes for juice clarification and the range of enzymes is being extended through new recombinant and non-recombinant fungal strains. Finally, novel fields of application can be envisaged for PGs in the production of oligogalacturonides as functional food components. Here we aim to highlight the various fields where PGs are encountered and where they are of biological or technological importance. PMID- 10803891 TI - Xylulose fermentation by mutant and wild-type strains of Zygosaccharomyces and Saccharomyces cerevisiae. AB - Anaerobic xylulose fermentation was compared in strains of Zygosaccharomyces and Saccharomyces cerevisiae, mutants and wild-type strains to identify host-strain background and genetic modifications beneficial to xylose fermentation. Overexpression of the gene (XKS1) for the pentose phosphate pathway (PPP) enzyme xylulokinase (XK) increased the ethanol yield by almost 85% and resulted in ethanol yields [0.61 C-mmol (C-mmol consumed xylulose)(-1)] that were close to the theoretical yield [0.67 C-mmol (C-mmol consumed xylulose)(-1)]. Likewise, deletion of gluconate 6-phosphate dehydrogenase (gnd1delta) in the PPP and deletion of trehalose 6-phosphate synthase (tps1delta) together with trehalose 6 phosphate phosphatase (tps2delta) increased the ethanol yield by 30% and 20%, respectively. Strains deleted in the promoter of the phosphoglucose isomerase gene (PGI1) - resulting in reduced enzyme activities - increased the ethanol yield by 15%. Deletion of ribulose 5-phosphate (rpe1delta) in the PPP abolished ethanol formation completely. Among non-transformed and parental strains S. cerevisiae ENY. WA-1A exhibited the highest ethanol yield, 0.47 C-mmol (C-mmol consumed xylulose)(-1). Other non-transformed strains produced mainly arabinitol or xylitol from xylulose under anaerobic conditions. Contrary to previous reports S. cerevisiae T23D and CBS 8066 were not isogenic with respect to pentose metabolism. Whereas, CBS 8066 has been reported to have a high ethanol yield on xylulose, 0.46 C-mmol (C-mmol consumed xylulose)(-1) (Yu et al. 1995), T23D only formed ethanol with a yield of 0.24 C-mmol (C-mmol consumed xylulose)(-1). Strains producing arabinitol did not produce xylitol and vice versa. However, overexpression of XKS1 shifted polyol formation from xylitol to arabinitol. PMID- 10803892 TI - Axenic cultivation of anoxygenic phototrophic bacteria, cyanobacteria, and microalgae in a new closed tubular glass photobioreactor. AB - A low-cost closed tubular glass photobioreactor allowing axenic cultivation of phototrophic microorganisms was constructed. Standard glass tubes were arranged in a helical array providing a working volume of 80 1. The glass tubes were connected with a degassing chamber, which also provided ports for measuring and regulating oxygen supply, pH, foam, and optical density and for adding substrates and antifoam agents as well as disposing of vent gas. A pump module allowed agitation of the medium in the bioreactor at a laminar flow rate of 1.5 m/s. Upstream of the pump module a gas inlet was located, allowing efficient mixing of the used gases with the medium. The temperature of the medium was controlled by a Pt-100 sensor and by a heat exchanger with an effective surface of 0.12 m2 connected to an external thermostat. Irradiation was provided by three light panels each consisting of ten fluorescent tubes. The entire photobioreactor - apart from the light panels and motor - could be sterilized at 121 degrees C in an autoclave. In addition to a detailed description of this photobioreactor, we report on first experiments to cultivate the anoxygenic phototrophic bacteria Rhodobacter sphaeroides and Rhodospirillum rubrum, the oxygenic phototrophic cyanobacterium Synechocystis sp. strain PCC6803, and the microalga Chlorella sp. in this photobioreactor. PMID- 10803893 TI - Soybean-milk-coagulating activity of Bacillus pumilus derives from a serine proteinase. AB - A proteolytic enzyme from Bacillus pumilus strain TYO-67, which was able to coagulate the protein in soybean milk, was characterized enzymologically. The optimum pH and temperature for its activities were 9.0 and 50 degrees C, respectively. The enzyme was strongly believed to be a serine proteinase because it was completely inhibited by the addition of diisopropyl fluorophosphate or phenylmethanesulfonyl fluoride. Hammerstein milk casein, cytochrome c and soybean protein were good substrates for the enzyme. Seven cleavages were detected using the oxidized insulin B-chain as peptide substrate for the proteolytic specificity test of the serine proteinase from B. pumilus. The bonds most susceptible to the action of the serine proteinase from B. pumilus were Leu-15-Tyr-16. The mode of action on soybean milk protein by the enzyme from B. pumilus was also investigated. The acidic subunit in glycinin and the alpha-, alpha- and beta subunits in beta-conglycinin were degraded during the enzyme reaction. However, the basic subunit in glycinin could not be degraded by the enzyme. The formation of coagula in soybean milk caused by the serine proteinase from B. pumilus was mainly due to the hydrophobic interaction. PMID- 10803895 TI - Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by recombinant bacteria expressing the PHA synthase gene phaC1 from Pseudomonas sp. 61-3. AB - Pseudomonas sp. 61-3 accumulated a blend of poly(3-hydroxybutyrate) [P(3HB)] homopolymer and a random copolymer consisting of 3-hydroxyalkanoate (3HA) units of 4-12 carbon atoms. The genes encoding beta-ketothiolase (PhbA(Re)) and NADPH dependent acetoacetyl-CoA reductase (PhbB(Re)) from Ralstonia eutropha were expressed under the control of promoters for Pseudomonas sp. 61-3 pha locus or R. eutropha phb operon together with phaC1(Ps) gene (PHA synthase 1 gene) from Pseudomonas sp. 61-3 in PHA-negative mutants P. putida GPp104 and R. eutropha PHB(-4) to produce copolyesters [P(3HB-co-3HA)] consisting of 3HB and medium chain-length 3HA units of 6-12 carbon atoms. The introduction of the three genes into GPp104 strain conferred the ability to synthesize P(3HB-co-3HA) with relatively high 3HB compositions (up to 49 mol%) from gluconate and alkanoates, although 3HB units were not incorporated at all or at a very low fraction (3 mol%) into copolyesters by the strain carrying phaC1Ps gene only. In addition, recombinant strains of R. eutropha PHB(-4) produced P(3HB-co-3HA) with higher 3HB fractions from alkanoates and plant oils than those from recombinant GPp104 strains. One of the recombinant strains, R. eutropha PHB(-4)/ pJKSc46-pha, in which all the genes introduced were expressed under the control of the native promoter for Pseudomonas sp. 61-3 pha locus, accumulated P(3HB-co-3HA) copolyester with a very high 3HB fraction (85 mol%) from palm oil. The nuclear magnetic resonance analyses showed that the copolyesters obtained here were random copolymers of 3HB and 3HA units. PMID- 10803894 TI - Metabolic engineering of carotenoid accumulation in Escherichia coli by modulation of the isoprenoid precursor pool with expression of deoxyxylulose phosphate synthase. AB - The recently discovered non-mevalonate pathway to isoprenoids, which uses glycolytic intermediates, has been modulated by overexpression of Escherichia coli D-1-deoxyxylulose 5-phosphate synthase (DXS) to increase deoxyxylulose 5 phosphate and, consequently, increase the isoprenoid precursor pool in E. coli. Carotenoids are a large class of biologically important compounds synthesized from isoprenoid precursors and of interest for metabolic engineering. However, carotenoids are not ordinarily present in E. coli. Co-overexpression of E. coli dxs with Erwinia uredovora gene clusters encoding carotenoid biosynthetic enzymes led to an increased accumulation of the carotenoids lycopene or zeaxanthin over controls not expressing DXS. Thus, rate-controlling enzymes encoded by the carotenogenic gene clusters are responsive to an increase in isoprenoid precursor pools. Levels of accumulated carotenoids were increased up to 10.8 times the levels of controls not overexpressing DXS. Lycopene accumulated to a level as high as 1333 microg/g dw and zeaxanthin accumulated to a level as high as 592 microg/ g dw, when pigments were extracted from colonies. Zeaxanthin-producing colonies grew about twice as fast as lycopene-producing colonies throughout a time course of 11 days. Metabolic engineering of carbon flow from simple glucose metabolites to representatives of the largest class of natural products was demonstrated in this model system. PMID- 10803897 TI - Cloning and characterization of an epoxide hydrolase-encoding gene from Rhodotorula glutinis. AB - We cloned and characterized the epoxide hydrolase gene, EPH1, from Rhodotorula glutinis. The EPH1 open reading frame of 1230 bp was interrupted by nine introns and encoded a polypeptide of 409 amino acids with a calculated molecular mass of 46.3 kDa. The amino acid sequence was similar to that of microsomal epoxide hydrolase, which suggests that the epoxide hydrolase of R. glutinis also belongs to the alpha/beta hydrolase fold family. EPH1 cDNA was expressed in Escherichia coli and resting cells showed a specific activity of 200 nmol min(-1)(mg protein)(-1) towards 1,2-epoxyhexane. PMID- 10803896 TI - His-tagged tryparedoxin peroxidase of Trypanosoma cruzi as a tool for drug screening. AB - Tryparedoxin peroxidase has recently been identified as a constituent of the complex peroxidase system in the trypanosomatid Crithidia fasciculata [Nogoceke E, Gommel DU, Kiess M, Kalisz HM, Flohe L (1997) Biol Chem 378: 827-836]. In trypanosomatids, hydroperoxides are reduced at the expense of NADPH by means of a cascade of three oxidoreductases: the flavoprotein trypanothione reductase, tryparedoxin and tryparedoxin peroxidase. Inhibitors of these enzymes are presumed to be trypanocidal drugs. Here, we present the heterologous expression of a putative tryparedoxin peroxidase gene of Trypanosoma cruzi (accession no AJ012101) as an N-terminally His-tagged protein (TcH6TXNPx). The product was purified with a high yield (8.75 mg from 11 fermentation broth of A(600)2.1) from the cytosolic fraction of sonified Escherichia coli BL21(DE3)[pET22b( + )/TcH6TXNPx] by metal-chelating chromatography. TcH6TXNPx proved to be fully active when tested with heterologous tryparedoxins of C. fasciculata (His-tagged TXN1H6 and TXN2H6). TcH6TXNPx displayed ping-pong kinetics with a k(cat) of 1.7 s(-1) and limiting Km values of 51.8 microM and 1.7 microM for t-butyl hydroperoxide and CfTXN2H6, respectively. PMID- 10803899 TI - Cloning of the cyclodextrin glucanotransferase gene from alkalophilic Bacillus sp. A2-5a and analysis of the raw starch-binding domain. AB - The cyclodextrin glucanotransferase (CGTase) gene of alkalophilic Bacillus sp. A2 5a was cloned and expressed in Bacillus subtilis ANA-1 as a host. The DNA region included an open reading frame encoding a 704-amino-acid polypeptide with a typical raw starch-binding motif in its C-terminal region. The CGTase purified from Bacillus sp. A2-5a bound to raw starch as strongly as porcine pancreas alpha amylase, as expected from the sequence motif. A chromosomal region (a DNA fragment of about 14.1 kbp) including the CGTase gene was also cloned and the nucleotide sequence was determined. Possible cyclodextrinase and putative cyclodextrin-binding protein genes were found in the flanking region of the CGTase gene, which implied that the novel starch-degradation pathway postulated for a gram-negative bacterium [Klebsiella oxytoca; Fiedler et al. (1996) J Mol Biol 256: 279-291] also exists in a gram-positive bacterium i.e. Bacillus. PMID- 10803898 TI - Sequence of PHA synthase gene from two strains of Rhodospirillum rubrum and in vivo substrate specificity of four PHA synthases across two heterologous expression systems. AB - A 3.0-kb genomic fragment has been isolated from Rhodospirillum rubrum (ATCC 25903) that contains an open reading frame (ORF) with strong homology to other known polyhydroxyalkanoate (PHA) synthase genes. This ORF has lower homology to the R. rubrum strain Ha PHA synthase than would be expected within the same species. We have conducted a series of heterologous expression studies evaluating the in vivo substrate specificity of PHA synthase genes from Rhodobacter sphaeroides, Ralstonia eutropha (formerly Alcaligenes eutrophus), Thiocystis violacea, and Nocardia corrallina, within the PHA-synthase-negative hosts, Ralstonia eutropha DSM541 and Pseudomonas putida GpP104. The N. corrallina PHA synthase incorporated the highest percentage of C5 monomers in the polymer when fermented in medium supplemented with 0.1% heptanoate as the sole carbon source. When the T. violacea and R. sphaeroides were expressed in the PHA-negative host DSM541, a greater percentage of C5 monomer was observed in the polymer as compared to the expression of the PHA synthase of R. eutropha, when the transconjugants were fermented in medium supplemented with 0.4% propionate. Evaluation for preference of medium-chain-length monomers demonstrated the flexibility of the N. corrallina, T. violacea, and R. eutropha synthase genes to polymerize a copolyester composed of short- and medium-chain-length monomers when the respective transconjugants were fermented in medium supplemented with 0.5% octanoate. These studies demonstrate that the PHA synthase from N. corrallina, T. violacea, and R. eutropha are able to polymerize a copolyester composed of short- and medium-chain-length monomers, while the PHA synthase from R. sphaeroides lacks this ability and only produces a short-chain-length polymer. These observations suggest that the composition of the PHA from the PHA-producing organisms does not necessarily reflect the inherent specificity of the PHA synthase. PMID- 10803900 TI - Propionic acid fermentation of glycerol and glucose by Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. shermanii. AB - A comparative study was carried out in anaerobic batch cultures on 20 g/l of either glycerol or glucose using two propionibacteria strains, Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. shermanii. In all cases, fermentation end-products were the same and consisted of propionic acid as the major product, acetic acid as the main by-product and two minor metabolites, n-propanol and succinic acid. Evidence was provided that greater production of propionic acid by propionibacteria was obtained with glycerol as carbon and energy sources. P. acidipropionici showed higher efficiency in glycerol conversion to propionic acid with a faster substrate consumption (0.64 g l(-1) h( 1)) and a higher propionic acid production (0.42 g l(-1) h(-1) and 0.79 mol/mol). The almost exclusive production of propionic acid from glycerol by this bacterium suggested an homopropionic tendency of this fermentation. Acetic acid final concentration was two times lower on glycerol (2 g/l) than on glucose (4 g/l) for both micro-organisms. P. freudenreichii ssp. shermanii exhibited a glycerol fermentation pattern typical of non-associated glycerol-consumption-product formation. This could indicate a particular metabolism for P. freudenreichii ssp. shermanii oriented towards the production of other specific components. These results tend to show that glycerol could be an excellent alternative to conventional carbon sources such as carbohydrates for propionic acid production. PMID- 10803901 TI - Formation and degradation of a synthetic humic acid derived from 3 fluorocatechol. AB - A synthetic fluorinated humic acid (FHA) was prepared by the spontaneous oxidative polymerization of 3-fluorocatechol. The 13C-solid-state NMR spectrum showed signals in the region for aromatic carbons with different substituents (aryl-H, aryl-C, aryl-O carbons) and for carboxyl-carbon. The latter indicated the formation of carboxylic groups, probably caused by ring cleavages during the polymerization process. An indication of the formation of carboxylic groups was also found in the infrared spectrum (band at 1715 cm(-1)). The dissolved FHA was degraded with active mycelium of the agaric white-rot fungus Nematoloma frowardii as well as with its isolated manganese peroxidase. In both cases, decolorization of the brownish FHA solution and partial defluorination (45-60%) took place. Degradation proceeded via formation of lower-molecular-mass fulvic acid-like substances. The results demonstrate that halogenated humic substances, e.g., resulting from the humification of xenobiotic compounds (bound residues), can in principle be eliminated by ligninolytic fungi (e.g., soil colonizing litter decomposers) and their manganese peroxidase system. PMID- 10803902 TI - Evidence for diverse oxidations in the catabolism of toluene by Rhodococcus rhodochrous strain OFS. AB - Rhodococcus rhodochrous strain OFS grew on toluene as a sole source of carbon and energy with a maximum growth rate of 0.011 h(-1). Initial reaction products were extracted, derivatized and identified by GC-MS. Oxygen consumption studies indicated that OFS grown on an aliphatic substrate required an induction period before oxidizing toluene. OFS grown on toluene transformed an array of aromatic ground water pollutants including styrene, ethylbenzene and chlorobenzene. Products of these transformations were identified. The sole product of chlorobenzene biotransformation was 4-chlorophenol. Products from toluene oxidation included 3- and 4-methylcatechol as well as benzyl alcohol, p-cresol and cis-toluene dihydrodiol. The identification of these and the products of other aromatic substrate conversions affirm that oxidation occurred on the functional group as well as directly on the aromatic nucleus. PMID- 10803903 TI - Polyhydroxyalkanoate accumulation in Burkholderia sp.: a molecular approach to elucidate the genes involved in the formation of two homopolymers consisting of short-chain-length 3-hydroxyalkanoic acids. AB - Burkholderia sp. accumulates polyhydrox-yalkanoates (PHAs) containing 3 hydroxybutyrate and 3-hydroxy-4-pentenoic acid when grown on mineral media under limited phosphate or nitrogen, and using sucrose or gluconate as a carbon and energy source. Solvent fractionation and NMR spectroscopic characterization of these polyesters revealed the simultaneous accumulation of two homopolyesters rather than a co-polyester with random sequence distribution of the monomers [Valentin HE, Berger PA, Gruys KJ, Rodrigues MFA, Steinbuchel A, Tran M, Asrar J (1999) Macromolecules 32: 7389-7395]. To understand the genetic requirements for such unusual polyester accumulation, we probed total genomic DNA from Burkholderia sp. by Southern hybridization experiments using phaC-specific probes. These experiments indicated the presence of more than one PHA synthase gene within the genome of Burkholderia sp. However, when total genomic DNA from Burkholderia sp. was used to complement a PHA-negative mutant of Ralstonia eutropha for PHA accumulation, only one PHA synthase gene was obtained resembling the R. eutropha type of PHA synthases, based on amino acid sequence similarity. In addition to the PHA synthase gene, based on high sequence homology, genes encoding a beta-ketothiolase and acetoacetyl-CoA reductase were identified in a gene cluster with the PHA synthase gene. The arrangement of the three genes is quite similar to the R. eutropha poly-beta-hydroxybutyrate biosynthesis operon. PMID- 10803904 TI - Simultaneous production of high activities of thermostable endoglucanase and beta glucosidase by the wild thermophilic fungus Thermoascus aurantiacus. AB - The culture-medium composition was optimised, on a shake-flask scale, for simultaneous production of high activities of endoglucanase and beta-glucosidase by Thermoascus aurantiacus using statistical factorial designs. The optimised medium containing 40.2 g l(-1) Solka Floc as the carbon source and 9 g l(-1) soymeal as the organic nitrogen source yielded 1130 nkat ml(-1) endoglucanase and 116 nkat ml(-1) beta-glucosidase activities after 264 h as shake cultures. In addition, good levels of beta-xylanase (3479 nkat ml(-1)) and low levels of filter-paper cellulase, beta-xylosidase, alpha-L-arabinofuranosidase, beta mannanase, beta-mannosidase, alpha-galactosidase and beta-galactosidase were detected. Batch fermentation in a 5-1 laboratory fermentor using the optimised medium allowed the production of 940 nkat ml(-1) endoglucanase and 102 nkat ml( 1) beta-glucosidase in 192 h. Endoglucanase and beta-glucosidase showed optimum activity at pH 4.5 and pH 5, respectively, and they displayed optimum activity at 75 degrees C. Endoglucanase and beta-glucosidase showed good stability at pH values 4-8 and 4-7, respectively, after a prolonged incubation (48 h at 50 degrees C). Endoglucanase had half-lives of 98 h at 70 degrees C and 4.1 h at 75 degrees C, while beta-glucosidase had half-lives of 23.5 h at 70 degrees C and 1.7 h at 75 degrees C. Alkali-treated bagasse, steam-treated wheat straw, Solka floc and Sigmacell 50 were 66, 48.5, 33.5 and 14.4% hydrolysed by a crude enzyme complex of T. aurantiacus in 50 h. PMID- 10803905 TI - Characteristics of glycosylated streptokinase secreted from Pichia pastoris: enhanced resistance of SK to proteolysis by glycosylation. AB - Degradation of streptokinase (SK) has been frequently observed during large-scale protein production. An enhanced susceptibility of SK to degradation has been correlated with its existence in a partially unfolded state. The influence of the carbohydrate moiety on the stability and functional characteristics of SK has been examined by obtaining the glycoform of SK following its secretion through the methylotrophic yeast Pichia pastoris. Secretion of the protein product was achieved by replacing the native secretion signal codons of SK with those from alpha-factor leader peptide and expressing the fusion construct under the control of the methanol-inducible alcohol oxidase (ox) promoter of P. pastoris after its integration into the host chromosome. Western blot and zymographic analysis of proteins secreted from the recombinant P. pastoris indicated that SK was glycosylated by the host cells, which resulted in the appearance of a SK species migrating slowly, corresponding to a 55-kDa protein product as compared to the 47 kDa native SK. The glycosylated SK retained a plasminogen activation capability identical to that of its unglycosylated counterpart. Glycoform SK exhibited an enhanced stability profile at 25 degrees C and 37 degrees C and improved resistance towards protease treatment compared to unglycosylated SK secreted through P. pastoris after tunicamycin treatment or that secreted from the recombinant Escherichia coli. The results presented thus illustrate that N-linked glycosylation of SK results in 30-40% enhancement of the protein stability and resistance towards degradation but does not interfere with its fibrinolytic function. PMID- 10803906 TI - Combined action of redox potential and pH on heat resistance and growth recovery of sublethally heat-damaged Escherichia coli. AB - The combined effect of redox potential (RP) (from -200 to 500 mV) and pH (from 5.0 to 7.0) on the heat resistance and growth recovery after heat treatment of Escherichia coli was tested. The effect of RP on heat resistance was very different depending on the pH. At pH 6.0, there was no significant difference, whereas at pH 5.0 and 7.0 maximum resistance was found in oxidizing conditions while it fell in reducing ones. In sub-lethally heat-damaged cells, low reducing and acid conditions allowed growth ability to be rapidly regained, but a decrease in the redox potential and pH brought about a longer lag phase and a slower exponential growth rate, and even led to growth failure (pH 5.0, < or =-100 mV). PMID- 10803907 TI - APHE-3, a spore-associated antibiotic of Streptomyces griseocarneus NCIMB 40447. AB - Solid cultures of the producing strain grown on Bennett medium develop abundant mycelium and intense sporulation. Under these conditions biosynthesis of APHE antibiotics (APHE-1 to -3) is accomplished. Further studies show that APHE-3 is basically produced during spore formation and mostly present in spores, while APHE-1 and APHE-2 are the predominant antibiotics in the mycelium. APHE compounds are present in almost all streptomycetes tested, indicating a possible role in the life cycle of these microorganisms. PMID- 10803908 TI - Dechlorination of polychlorinated methanes by a sequential methanogenic denitrifying bioreactor system. AB - A two-stage bioreactor has been developed to link dechlorination of halogenated methane compounds to the anaerobic processes of methanogenesis and denitrification. A digester methanogenic consortium was shown to dechlorinate chloroform (CF) and carbon tetrachloride (CT) to dichloromethane (DCM), and DCM was then mineralized by an acclimated denitrifying biological activated carbon consortium. Combining these two processes, a sequential methanogenic-denitrifying bioreactor (SMDB) system that completely degraded polychlorinated methanes including CT, CF, and DCM was developed. More than 95% of the added CT and CF was dechlorinated in the methanogenic bioreactor with methanol as the primary substrate, and the resultant DCM was biodegraded in the denitrifying bioreactor with nitrate as the electron acceptor. In the denitrifying bioreactor, the residual CF was completely removed, and the DCM removal efficiency was more than 95%. This novel bioreactor system eliminates the need for aeration and so avoids the air contamination associated with aerobic biotreatment of volatile chlorinated pollutants. This SMDB system provides an alternative to conventional biotreatment of wastewaters and other matrices contaminated with polychlorinated methanes and is, to our knowledge, the first report on such a sequential anoxic system. PMID- 10803909 TI - Auditory deprivation of the central auditory system resulting from selective inner hair cell loss: animal model of auditory neuropathy. AB - Auditory neuropathy is often characterized by normal thresholds, present otoacoustic emissions, poor speech discrimination, absent acoustic reflexes, absent or abnormal auditory brainstem response waveform, but normal late cortical potential. This paper describes an animal model that has many characteristics of auditory neuropathy. Chinchillas can be deprived of a significant portion of the neural inputs to the central auditory system by administering carboplatin, an antineoplastic agent that selectively destroys inner hair cells (IHCs) and type I auditory nerve fibers. Selective IHC loss has no effect on distortion product otoacoustic emissions or the cochlear microphonic potential, implying normal outer hair cell function. However, selective IHC loss causes the amplitude of the compound action potential to decrease in proportion to the degree of IHC loss. However, the threshold of the CAP shows little increase with mild to moderate IHC loss. Acoustically responsive auditory nerve fibers in ears with mild to moderate IHC loss have normal thresholds and tuning curves with narrowly tuned tips. Although the central auditory pathway is deprived of much of its sensory inputs, the amplitude of the local field potential in the auditory cortex was normal or enhanced, while those from the inferior colliculus were slightly reduced. The results are related to those of a patient with auditory neuropathy. PMID- 10803910 TI - Prolonged deafness limits auditory system developmental plasticity: evidence from an evoked potentials study in children with cochlear implants. AB - The use of cochlear implants to restore hearing in profoundly deaf children is increasing, with a trend toward earlier implantation. However, little is known about how auditory deprivation and subsequent implant use affects the maturing central auditory system. Previously reported results indicate that stimulation of the auditory system by a cochlear implant is sufficient to restore at least some aspects of central auditory pathway maturation, as reflected by age-related changes in the auditory evoked potentials. We review animal and human studies on sensory deprivation and report new results based on longitudinal evoked potentials data recorded from two individuals. Analyses show that age-related changes in the EPs may asymptote at levels very different from those found in the adult normal-hearing population. These results suggest that maturation of at least some aspects of central auditory system activity is limited by the onset and duration of the period of deafness prior to implantation. PMID- 10803911 TI - Generation of auditory steady-state responses: linearity assessment. AB - Auditory steady-state responses (SSRs) are believed to result from the superimposition of the middle latency responses (MLRs) evoked by individual stimuli. Our recent studies challenge this hypothesis in several regards. Surface electrical recordings performed in 16 normal subjects showed that the prediction curves obtained by MLR linear addition failed to predict SSRs at rates other than 40 Hz. Amplitude and phase differences between actual and predicted responses point to the intervention of phenomena related to the recovery cycle of the neural networks underlying the SSR generation. A click train paradigm at a 40 Hz rate was utilized and an approximation to the response evoked by the last stimulus was obtained by an analytical handling. The most relevant feature of this response in comparison to the MLR was the appearance of an additional activity which could be related to the fast oscillations of auditory cortical neurons. Our findings suggest that the responses evoked by individual stimuli during steady-state stimulation change by increasing the repetition rate, thus contradicting the most widely accepted hypothesis of the MLR linear addition in the SSR generation. PMID- 10803912 TI - Central auditory tests. AB - This article overviews some selected central auditory test procedures as well as some popular issues that surround them. Both behavioral and electrophysiologic central auditory tests are discussed in reference to their clinical value as judged by their sensitivity, specificity, administration time, and reliability. Also discussed are diagnostic trends that are noted on behavioral and electrophysiologic procedures. Relationships are drawn between children with auditory processing deficits associated with learning problems and individuals with neurological lesions of the central auditory nervous system. PMID- 10803913 TI - Neuroanatomy of tinnitus. AB - We tested the hypothesis that tinnitus was due to excessive spontaneous activity in the central auditory system by seeking cerebral blood flow (CBF) changes that paralleled changes in the loudness of tinnitus in patients able to alter the loudness of their tinnitus. We found CBF changes in the left temporal lobe in patients with right ear tinnitus, in contrast to bilateral temporal lobe activity associated with stimulation of the right ear. The tones activated more extensive portions of the brain in patients than controls. We conclude that tinnitus is not cochlear in origin and associated with plastic transformations of the central auditory system. We suggest that tinnitus arises as a consequence of these aberrant new pathways and may be the auditory system analog to phantom limb sensations in amputees. PMID- 10803914 TI - Clinical relevance of medial efferent auditory pathways. AB - Evoked otoacoustic emissions have been shown to be suppressed by sounds applied in the contralateral ear and this effect can be largely explained by the involvement of medial olivocochlear efferent fibers. Thus, EOAE recording during contralateral stimulation provides a non-invasive means of investigating auditory efferent system functioning in humans. The question remains, however, as to whether this test provides a tool, which could be useful in a variety of clinical applications. This review describes current clinical applications for this test, showing that it may prove useful for improving identification of retrocochlear pathologies. Some new areas are also identified. Methodological topics are discussed and suggestions for maximizing the value of this test are proposed. PMID- 10803915 TI - Selected management approaches to central auditory processing disorders. AB - This is a commentary on three key aspects of the management of central auditory processing disorders in children. It is an update on some selected approaches for the management of auditory processing problems. Auditory training techniques that are theoretically based and for which empirical data are emerging are addressed. The second part of our commentary deals with methods of enhancing acoustic speech signals. Discussions on clear speech, acoustic modifications of the listening environment, and the utilization of assistive listening devices are presented. The final part of the article reviews linguistic and cognitive interventions for people with auditory processing problems. Topics in this section include approaches to facilitate phonological awareness, vocabulary building, prosodic feature recognition, and use of metacognitive abilities. PMID- 10803916 TI - The medial olivocochlear efferent system in humans: structure and function. AB - The mammalian cochlea receives innervation from the central nervous system via two efferent systems: the lateral and the medial olivocochlear bundles. Advances in cochlear physiology have clarified the origins and terminations of these fibers. However, to date, while the functional significance of lateral efferents remains totally unknown, that of medial efferents is still controversial. The peripheral effects of medial olivocochlear system activation have been described, but the role of this inhibitory feedback onto the cochlea is still unclear. This overview summarizes the main results of several psychophysiological studies performed in humans dealing with the functional significance of medial olivocochlear efferents in hearing. PMID- 10803917 TI - Modulation of plasma uridine concentration by 5-(phenylselenenyl)acyclouridine, an inhibitor of uridine phosphorylase: relevance to chemotherapy. AB - PURPOSE: The purpose of this investigation was to evaluate the efficacy of oral 5 (phenylselenenyl)-acyclouridine (PSAU) in increasing endogenous plasma uridine concentration as well as its ability to improve the bioavailability of oral uridine. PSAU is a new potent and specific inhibitor of uridine phosphorylase (Urd-Pase, EC 2.4.2.3), the enzyme responsible for uridine catabolism. This compound was designed as a lipophilic inhibitor in order to facilitate its access to the liver and intestine, the main organs involved in uridine catabolism. METHODS: Oral PSAU was administered orally to mice alone or with uridine. The plasma levels of PSAU as well as uridine and its catabolites were measured using high-performance liquid chromatography and pharmacokinetic analysis was performed. RESULTS: PSAU has an oral bioavailability of 100% and no PSAU metabolites were detected. PSAU has no apparent toxicity at high doses. Oral administration of PSAU at 30 and 120 mg/kg increased baseline concentration of endogenous plasma uridine (2.6 +/- 0.7 microM) by 3.2- and 8.7-fold, respectively, and remained three- and six-fold higher, respectively, than the controls for over 8 h. PSAU, however, did not alter the concentration of endogenous plasma uracil. Co-administration of PSAU with uridine elevated the concentration of plasma uridine over that resulting from the administration of either alone, and reduced the peak plasma concentration (C(max)) and area under the curve (AUC) of plasma uracil. Co-administration of PSAU at 30 mg/kg and 120 mg/kg improved the low bioavailability of oral uridine (7.7%) administered at 1,320 mg/kg by 4.8- and 4.2-fold, respectively, and reduced the AUC of plasma uracil from 1,421 to 787 micromol/h x l and 273 micromol/h x l, respectively. Similar results were observed when PSAU was co-administered with lower doses of uridine. Oral PSAU at 30 mg/kg and 120 mg/kg improved the bioavailability of oral 330 mg/kg uridine by 5.2- and 8.9-fold, and that of oral 660 mg/kg uridine by 6.4 and 9.0-fold, respectively. However, the reduction in the AUC values of plasma uracil was less dramatic than that seen when the high dose of 1,320 mg/kg uridine was used. CONCLUSION: The effectiveness of the PSAU plus uridine combination in elevating and sustaining high plasma uridine concentration may be useful to rescue or protect from host toxicity of various chemotherapeutic pyrimidine analogs as well as in the management of medical disorders that are remedied by administration of uridine. PMID- 10803918 TI - P53 status plays no role in radiosensitizing effects of SN-38, a camptothecin derivative. AB - PURPOSE: Topoisomerase inhibitors including camptothecin are being studied as potential radiosensitizers. CPT-11 is a derivative of camptothecin and is clinically available. In this study, we investigated the effects of SN-38 (an active metabolite of CPT-11) on four nonirradiated and irradiated murine fibroblast cell lines with different p53 statuses to clarify the role of p53 in the radiosensitizing activity of SN-38. MATERIALS AND METHODS: Four fibroblast cell lines, MT158, MT158/neo, MT158/wtp53 and MT158/mp53 with the same genetic background but with different p53 statuses, were used. Exponentially growing cells were treated with SN-38 (200 nM) and incubated with the drug for 30 min. Cells were then irradiated (0 to 12 Gy) and further incubated with the drug for 2 h. The cell survival rate was determined using a conventional clonogenic assay. The effects of the treatments on the cell cycle were analyzed with a flow cytometric assay. Apoptosis after these treatments was also detected by an annexin V assay. RESULTS: There were no significant differences in sensitivity to radiation or SN-38 treatment among these cell lines. The combined treatment of irradiation and SN-38 showed supraadditive effects in all four cell lines independent of their p53 status. Transient arrest in G2 with a decreased percentage of cells in both the S and G1 phases was observed 8 h after treatment with either SN-38 alone, radiation or their combination, regardless of the p53 status. No significant differences in frequency of apoptosis were observed between treatment and control groups in two cell lines with or without wild-type p53. CONCLUSION: The combination of irradiation and SN-38 treatment showed supraadditive effects in all four cell lines tested, and the p53 status did not play a role in the combination effect. PMID- 10803919 TI - The role of p53 in gemcitabine-mediated cytotoxicity and radiosensitization. AB - PURPOSE: We compared the cytotoxic and radiosensitizing effects of gemcitabine (2',2'-difluoro-2'-deoxycytidine, dFdCyd), a clinically valuable radiosensitizer, in colon cancer RKO cells which differed in their p53 status. The parental RKO cells, RKO-P, contain wild-type p53 protein. In RKO-E6 cells, the p53 function has been disrupted by transfection of the cells with the human papillomavirus type-16 E6 gene. RESULTS: We found that the RKO-P cells were significantly more sensitive to dFdCyd-mediated cytotoxicity and apoptosis than RKO-E6 cells (IC10 39.3 +/- 5.3 nM and 62.0 +/- 6.9 nM, respectively). The cytotoxic effect of dFdCyd in RKO-P cells was accompanied by induction of the proapoptotic protein Bax at the time when p53 was induced. In contrast, similar treatment of RKO-E6 cells with dFdCyd resulted in only limited expression of Bax, suggesting that the cytotoxic effect of dFdCyd was mediated, in part, by a p53-dependent apoptosis pathway. We also studied the effect of dFdCyd on radiation sensitivity. We found that at minimally cytotoxic concentrations dFdCyd failed to radiosensitize either RKO-P or RKO-E6 cells, whereas at cytotoxic concentrations equal sensitization was produced. Finally, we assessed the influence of dFdCyd on cell cycle distribution. We found that dFdCyd synchronized RKO-P cells, whereas synchrony was not produced in p53-disrupted RKO-E6 cells. CONCLUSION: These results suggest that p53 status may influence dFdCyd-mediated apoptosis, cytotoxicity, and cell cycle progression but do not support an important role for p53 in radiosensitization. PMID- 10803920 TI - In vivo measurement of myocardial oxidative metabolism and blood flow does not show changes in cancer patients undergoing doxorubicin therapy. AB - PURPOSE: The aim was to investigate in patients receiving doxorubicin whether any alteration in myocardial oxidative metabolism or blood flow as assessed by positron emission tomography (PET) could be observed either after the first dose of the drug, or during its chronic administration. METHODS: Six female non-heart failure cancer patients treated with doxorubicin were included in a longitudinal study. Resting radionuclide cineangiography and PET scanning with carbon-11 acetate were performed the day before the initiation of doxorubicin treatment at a dosage of 50 mg/m2 every 3 weeks, and 3 weeks after the cumulative administration of 300 mg/m2 (chronic toxicity). In addition, PET was performed 24 h after the first administration of doxorubicin (evaluation of acute toxicity). Myocardial oxidative metabolism and blood flow were assessed by PET (acute and chronic toxicity), and left ventricular ejection fraction was measured by radionuclide angiography (chronic toxicity). RESULTS: Using PET for both acute and chronic toxicity evaluations, no significant effect of doxorubicin was observed either on the flux through the tricarboxylic acid (TCA) cycle or on myocardial blood flow. However, systolic left ventricular function showed a small but significant impairment after the administration of 300 mg/m2 of doxorubicin. CONCLUSIONS: Other hypotheses should be explored to better explain the predominant mechanisms of the cardiotoxicity of anthracyclines in humans. PMID- 10803921 TI - Increase in tumor GADD153 mRNA level following treatment correlates with response to paclitaxel. AB - PURPOSE: We investigated the relationship between the basal and treatment-induced change in the tumor expression of the drug resistance gene MDR1 and the cellular injury response gene GADD153, and clinical response to paclitaxel treatment. METHODS: MDR1 and GADD153 mRNA levels were measured by reverse transcriptase polymerase chain reaction (RT-PCR) in tumor samples obtained by fine needle aspiration biopsy from 14 patients before and 24 h after paclitaxel infusion. RESULTS: There was no difference between responders and non-responders with respect to either the basal MDR1 mRNA level or the change in MDR1 mRNA level at 24 h after treatment (P = 0.464). Likewise, there was no difference in basal GADD153 mRNA level between responders and non-responders. However, there was a significantly greater increase in GADD153 mRNA at 24 h in responders compared with non-responders (P = 0.005). An increase in GADD153 mRNA level of 1.5-fold or higher predicted response with a sensitivity of 86% and a specificity of 100%. CONCLUSIONS: An increase in GADD153 mRNA level reflects chemotherapy-induced damage sufficient to be manifest as a clinically detectable reduction in tumor volume. Measurement of the change in GADD153 mRNA level successfully identified patients destined to respond as early as 24 h post-treatment. PMID- 10803922 TI - Selective inhibition of cyclooxygenase-2 enhances mitomycin-C-induced apoptosis. AB - PURPOSE: Cyclooxygenase-2 (COX-2) is involved in antiapoptosis signaling, and its induction may require activation of protein kinase C (PKC). Safingol (SAF), a PKC inhibitor, has been shown to enhance apoptosis induced by mitomycin-C (MMC) in human gastric cancer MKN-74 cells. The aim of this study was to identify the role of COX-2 in MMC-induced apoptosis in MKN-74 cells. METHODS: Protein expression of COX-2 and Bcl-2 and activation of PKCalpha were examined by Western blot analysis. Apoptosis induction was examined by staining with bisbenzimide trihydrochloride (Hoechst-33258) of condensed chromatin, which characterizes the cells undergoing apoptosis. COX-2 mRNA levels were examined by Northern blot analysis. RESULTS: After exposure for 1-2 h to 1 microg/ml MMC, upregulation of COX-2 and Bcl-2 protein expression was noted. The activation of PKCalpha occurred within 1 h of MMC exposure, and temporally preceded the induction of COX-2. Similar results were observed in cells exposed to the PKC activator, 3-phorbol 12 myristate 13-acetate. Cotreatment with SAF and MMC abolished the induction of COX 2 by MMC. Furthermore, NS-398, a selective COX-2 inhibitor, significantly enhanced MMC-induced apoptosis by fivefold from 4 +/- 2% (MMC alone) to 20 +/- 2% (MMC plus NS-398). There was no discernible change in COX-2 mRNA levels after a 2 h exposure to MMC but a twofold increase after a 24-h exposure. CONCLUSIONS: MMC upregulates COX-2 expression, which appears to be an antiapoptotic signal downstream of PKC. Selective inhibition of COX-2 can therefore provide a novel way to enhance MMC-induced apoptosis independent of inhibiting PKC. PMID- 10803923 TI - Evaluation of antidotes for extravasation injury produced by 6 hydroxymethylacylfulvene (MGI 114), a novel cytotoxic antitumor agent, in an intradermal toxicity model in rats. AB - MGI 114 (HMAF, 6-hydroxymethylacylfulvene) is a cytotoxic drug currently in phase II human clinical trials. As with other anticancer agents, inadvertent drug extravasation may result in perivascular irritation and/or necrosis. In this study the degree of soft tissue injury produced by MGI 114 after intradermal administration to rats was quantified and four potential antidotes for extravasation injuries caused by MGI 114 were evaluated. Intradermal injections of MGI 114 (0.2 ml, concentrations 0.1, 0.5 or 1.0 mg/ml) and a positive control, doxorubicin (0.2 ml, concentration 2 mg/ml) were administered to male Fischer 344 rats in an experiment designed to establish a model for antidote evaluation. Dermal lesions at the injection sites were measured and quantitated as the total area under the lesion area-time curve (AUC). Physiological saline, sodium thiosulfate, dimethylsulfoxide (DMSO) and local cooling, were then compared as potential antidotes in this model. In the initial study, dermal lesions (erythema, ulcerations and eschar formation) occurred at the MGI 114- and doxorubicin-treated sites. The lesion area resulting from MGI 114 was dose related and was greatest at approximately 5 days, with resolution by day 7-22. Doxorubicin-induced lesions were comparable in area to those induced by the highest dose of MGI 114, but persisted approximately twice as long. In the antidote study, sodium thiosulfate administration resulted in approximately 20% diminution of lesion area and AUC value when compared to untreated controls. Normal saline caused slight reductions in maximum lesion area, but had little effect on AUC values. Local cooling also caused a modest reduction in the maximum lesion area, but actually resulted in higher AUC values by prolonging eschar duration. DMSO provided near complete tissue protection from intradermal exposure to MGI 114. In this model MGI 114 and doxorubicin were found to produce similar soft tissue injuries, but MGI 114-induced lesions tended to show a more rapid resolution. Topical DMSO treatment was found to produce the most effective protection against MGI 114-induced local tissue irritation and necrosis. PMID- 10803924 TI - Dose-dependent hormonal effects of toremifene in postmenopausal breast cancer patients. AB - PURPOSE: The purpose of the study was to compare hormonal effects of three toremifene doses, 20 mg (TOR20), 40 mg (TOR40) and 60 mg (TOR60) administered daily, in postmenopausal women with advanced breast cancer. METHODS: The study was randomized and open label in three parallel groups. Biochemical variables were identified as the serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH) and sex hormone binding globulin (SHBG). The changes were compared with objective clinical responses and to progression-free time. Adverse reactions and liver function test (aspartate aminotransferase, ASAT) were assessed for safety. RESULTS: A total of 260 patients were randomly grouped (90 to TOR20, 81 to TOR40 and 89 to TOR60). Of these patients 29, 29 and 22 completed at least 3 months of treatment and the results were analyzed for biochemical variables. All treatments had intrinsic estrogen agonist activity by decreasing of serum FSH and LH and by increasing of SHBG during the first 3 months (P < 0.01). Dose TOR20 showed slightly longer times to exert maximum estrogenic effects than did the two higher doses. No increases in liver function tests were seen in any of the groups. Objective response rates were 24.4, 39.5 and 32.6% (P = 0.01) and median times-to-progression were 206, 189 and 196 days in TOR20, TOR40 and TOR60, respectively (P = 0.913). Fewer responses were observed in the TOR20 group than in TOR40 (P = 0.05). Adverse events were reported in 19, 23 and 30 patients in the treatment groups (P = 0.20). The most frequently reported events were hot flushes and nausea. These were mostly mild or moderate, and only 1.5% of treatments was discontinued due to toxicity. CONCLUSIONS: Toremifene doses of 40 and 60 mg daily were effective and safe treatments of breast cancer in postmenopausal women, and no differences in their biochemical or clinical effects were seen. Toremifene at 20 mg/day had similar but slightly less potent antiestrogenic and estrogenic effects than the two higher doses. PMID- 10803925 TI - Identification and proposed mechanism of action of thymidine kinase inhibition associated with cellular exposure to camptothecin analogs. AB - PURPOSE: To investigate the effects of several camptothecin analogs including 9 aminocamptothecin (9-AC), SN38, topotecan, and irinotecan (CPT-11) on the enzymes involved in the pyrimidine salvage pathway including thymidylate synthase (TS). A COMPARE analysis using the NCI 60 cell line drug-screening panel suggested that there were similarities in the mechanisms of action of camptothecin analogs and TS inhibitors. METHODS: TS enzymatic activity was measured by both an in situ tritium release assay using both the H630 colon cancer cell line and the CEM human leukemia cell line, and by a radiolabelled in vitro assay using partially purified human TS as the enzyme source. Thymidine kinase (TK) activity was measure by a radiolabelled in vitro assay using H630 colon cancer cell lysates as the enzyme source. RESULTS: In vitro studies indicated that none of the analogs directly inhibited TS enzymatic activity; however, utilization of a coupled TS/TK in situ assay with radiolabelled deoxyuridine as the precursor revealed marked inhibition by the camptothecin analogs. 9-AC, SN38, and topotecan yielded IC50 values of 1.3, 1.6, and 1.1 microM respectively. In contrast, there was no inhibition detected when deoxycytidine was used as the radiolabelled nucleoside precursor, suggesting that the drug effect was through inhibition of TK, rather than inhibition of TS. In vitro studies using cell lysates from H630 human colon cancer cells to measure TK activity showed no decrease in TK activity after 9-AC treatment. In addition, no changes were detected in the dATP and dTTP nucleotide pools. Permeabilizing the cell membranes with saponin did not abolish the inhibitory effect of the camptothecins indicating that altered cell transport was not responsible for the decreased activity in the in situ assay in intact cells. CONCLUSION: These studies suggest that there is inhibition of TK in intact cells associated with topoisomerase I inhibition by camptothecin analogs, and the inhibition of TK is the result of an indirect effect not related to feedback inhibition by changes in dTTP pools. PMID- 10803926 TI - Effects of protein binding on the in vitro activity of antitumour acridine derivatives and related anticancer drugs. AB - PURPOSE: We set out to measure drug binding to serum proteins. These have been shown to reduce the free plasma concentrations of a number of anticancer drugs, particularly of those of complex organic structure, in both experimental studies and clinical trials. METHODS: We have used cultures of murine Lewis lung carcinoma cells as sensors of available drug to measure the effects of two drug binding plasma proteins, alpha-acid glycoprotein (AAG) and human serum albumin (HSA), as well as of bovine serum albumin (BSA) on drug activity. RESULTS: The concentrations required for 50% growth inhibition (IC50 values) of a number of anticancer drugs were found to be linear functions of the added proteins. Assuming that cells respond to free drug, the data provide estimates of the product K x n, where K is the binding constant of the protein and n is the number of drug binding sites per protein molecule. Amsacrine, the amsacrine analogue asulacrine, camptothecin, DACA (N-[2-(dimethylamino)ethyl]acridine-4 carboxamide), doxorubicin, etoposide, mitoxantrone, paclitaxel and vincristine were tested. The K x n values for AAG were 30, 2,400, 8.7, 340, 29, 290 x 10(3) M(-1) and 120 x 10(3) M(-1), respectively, and the K x n values for HSA were 16, 580, 530, 10, 6.2, 4.3 x 10(3) M(-1) and 0.0 x 10(3) M(-1), respectively. The combined data allowed the estimation of free fractions of drug in plasma, assuming that AAG and HSA contributed most to protein binding. The data were in general comparable with that reported using equilibrium dialysis and ultrafiltration. Data for drug binding to BSA were different from those for HSA, in some cases by a large factor with values for HSA generally higher. The applicability of the method to analogue development was illustrated by examining the binding to AAG of a series of DACA analogues, and binding was found to be primarily related to lipophilicity. CONCLUSION: IC50 determinations provide a rapid means of estimating drug binding to plasma proteins and have utility in the assessment of new anticancer drugs. PMID- 10803927 TI - Phase I study of AG2034, a targeted GARFT inhibitor, administered once every 3 weeks. AB - PURPOSE: To identify a recommended phase II dose for the second generation glycinamide ribonucleotide transformylase (GARFT) inhibitor, AG2034, administered by intravenous bolus every 3 weeks without folate supplementation and to describe AG2034 pharmacokinetics. METHODS: Adults with advanced malignancies were enrolled in cohorts of three per dose level with expansion to six upon observation of dose limiting toxicity (DLT). The maximum tolerated dose (MTD) was defined as the dose at which two of up to six patients experienced DLT. Upon identification of an MTD and evidence of cumulative toxicity, a lower intermediate dose was explored as a candidate phase II dose. AG2034 plasma concentrations were measured using an ELISA assay. RESULTS AND CONCLUSIONS: The recommended phase II dose is 5.0 mg/m2. DLTs were anemia, thrombocytopenia, mucositis, diarrhea, hyperbilirubinemia, fatigue, and insomnia. Toxicities were modestly cumulative over three courses. Pharmacokinetic analysis showed a dose-AUC0-24 relationship and a progressive increase in AG2034 AUC0-24 over three courses. Both pharmacokinetic and pharmacodynamic factors may contribute to the modest cumulative toxicity observed with AG2034. PMID- 10803928 TI - Successful treatment with paclitaxel of a patient with metastatic extra-adrenal pheochromocytoma (paraganglioma). A case report and review of the literature. AB - This case report describes the history of a patient with an aggressive course of a metastatic extra-adrenal pheochromocytoma (paraganglioma) who received different combination chemotherapy regimens with no or short-lasting clinical benefit. However, during treatment with single-agent paclitaxel, there was a significant clinical improvement, a partial biochemical response and a minor roentgenologic response, which was sustained for 1 year. In this report we present this case and also review the literature on the chemotherapy used for this rare disease over the past 15 years. To enable the activity of paclitaxel against this neoplasm to be determined, more patients need to be treated. PMID- 10803929 TI - Nasal absorption of ondansetron in rats: an alternative route of drug delivery. AB - PURPOSE: Ondansetron (OND) is a 5-HT3 receptor antagonist that is used therapeutically for the prevention of nausea and vomiting associated with emetogenic cancer therapy. There is a need for nasal drug delivery in specific patient populations where the use of commercially available intravenous and oral dosage forms may be inconvenient and/or unfeasible. METHODS: OND (Zofran Injection, 2 mg/ml) was administered at a dose of 1 mg/kg to male Sprague-Dawley rats intravenously or intranasally (n = 3 in each group). A special surgical procedure was performed to ensure that the drug solution was held in the nasal cavity. OND was injected into the femoral vein for the intravenous group. Blood samples were collected at appropriate times for 60 min. An HPLC method was employed to determine OND in the plasma. RESULTS: The results clearly showed that OND was readily and rapidly absorbed through the nasal mucosa of the rat. The peak plasma level was attained within 10 min. OND was also completely absorbed as the plasma concentration-time profiles for the two routes were comparable. The terminal elimination half-lives were also similar. CONCLUSIONS: The nasal administration route for OND is apparently as effective as the intravenous route. If one considers the limitations of delivering OND orally or intravenously to patients undergoing emetogenic cancer therapy, it becomes obvious that the intranasal route is a potential alternative modality to prevent nausea and vomiting associated with such therapy. PMID- 10803930 TI - Interferon alpha in the treatment of polycythemia vera. AB - Interferon alpha (IFN) inhibits the growth of the abnormal clone in patients with myeloproliferative disorders, leading to a reduction of the clinical and laboratory signs of the pathologic myeloproliferation. The therapeutic efficacy of IFN in polycythemia vera (PV) is demonstrated by the summarized treatment results of 279 patients participating in 16 prospective nonrandomized studies and in three case reports. The initial IFN dose ranged from 3 to 35 million IU/week. In 82% of the patients the frequency of phlebotomies was reduced. In 50% a complete remission was achieved, defined as a stable hematocrit of 45% without concomitant phlebotomies. Reduction of splenomegaly was seen in 77% and control of pruritus in 81% of the patients. The median observation time of the studies was 13 months (ranging from 3 to 84 months). Individual cases were followed for up to 126 months. In 21% of the patients IFN was terminated, owing mostly to side effects. The selective suppression of the malignant clone by IFN was demonstrated by the induction of cytogenetic remissions in sporadic cases with a chromosomal marker and by the observation of unmaintained remissions that lasted up to 4.8 years. IFN has no known mutagenic or teratogenic effects. The data presently available demonstrate that IFN is an effective alternative to the present forms of treatment in PV. Controlled prospective studies are essential to clarify whether the favorable biologic properties are also reflected by a benefit in clinical course and survival, and whether IFN may reduce the rates of acute leukemia and myelofibrosis. A randomized study that compares IFN and hydroxyurea in patients with PV has recently been initiated by the Suddeutsche Hamoblastosegruppe (SHG) in Germany. PMID- 10803931 TI - Poor clinical outcome of patients with Hodgkin's disease and elevated interleukin 10 serum levels. Clinical significance of interleukin-10 serum levels for Hodgkin's disease. AB - Interleukin (IL)-10 is a pleiotropic cytokine with potent inhibitory effects towards T(H)-1 cells. IL-10 inhibits secretion of IL-2 and interferon (IFN)gamma by T cells and downregulates major histocompatibility complex antigens. A variety of tumor cells secrete IL-10, which can inhibit growth of tumor-specific cytotoxic T cells. IL-10 expression has also been detected in B-cell lymphomas and Hodgkin's disease (HD), and it has been suggested that the cytokine is involved in the pathogenesis of these tumors. We analyzed levels of IL-10 in pretreatment sera of 64 patients with HD and healthy controls using a sensitive enzyme-linked immunosorbent assay. Patients with biopsy-proven HD were enrolled in trials of the German Hodgkin Study Group (GHSG). Elevated IL-10 levels were detected in the sera of nine patients with HD (14.1%) (range 4.5-225.6 pg/ml with a mean of 61.5 pg/ml). IL-10 was not detectable in a control population of healthy volunteers (n =90). Multivariate analyses revealed a significant correlation between elevated IL-10 levels and higher age (over 45 years) but not with any other factors defined by the international prognostic factor score. Patients with elevated IL-10 levels had a significantly lower freedom from treatment failure rate as detected in univariate and multivariate tests. Thus, IL 10 may serve as an independent prognostic factor for HD patients. PMID- 10803932 TI - Thyroid toxicity of treatment for Hodgkin's disease. AB - Thyroid disease, especially hypothyroidism, is one of the more frequently encountered late endocrine sequelae of treatment for Hodgkin's disease. We analyzed the thyroid function of 177 patients (92 male and 85 female) who had been treated for Hodgkin's disease between 1970 and 1995; their median age was 38 years (range 18-74) and their median time after therapy was 6 years (range 1-20). Thirty-five (20%) patients were treated with chemotherapy alone (mainly COPP/ABVD), 44 (25%) with radiotherapy (RTX) alone, and 98 (55%) received combined modality treatment according to the protocols of the German Hodgkin's Disease Study Group. All patients had been without evidence of disease for at least 1 year. They were evaluated between 1994 and 1997 for thyroid disease by clinical examination, thyroid function tests, and ultrasound imaging if indicated. Overall, 48 patients (27%) were found to have subclinical (36 patients, 20%) or overt (12 patients, 7%) hypothyroidism. No patient in the group treated with chemotherapy alone developed hypothyroidism, in contrast to 15 of 44 (34%) patients treated with radiotherapy alone and 33 of 98 (34%) patients treated with the combined modality approach who did. All patients with hypothyroidism had received some form of supradiaphragmatic radiation except one patient who underwent infradiaphragmatic RTX but with preceding hemithyroidectomy for staging purposes. After an average follow-up of 6 years, 27% of patients treated for Hodgkin's disease with either radiotherapy alone or a combination of radiotherapy and chemotherapy developed hypothyroidism. Supradiaphragmatic radiation, but not the use of chemotherapy alone, is associated with an increased risk for hypothyroidism. Evaluation of thyroid function after therapy is important, and appropriate substitution medication should be initiated. PMID- 10803933 TI - The activation of intracellular tyrosine kinases by interferon-alpha (IFNalpha) correlates with its antiproliferative activity in B-lymphoid cell lines, but not in B-cell chronic lymphocytic leukemia patients. AB - The response to interferon-alpha (IFNalpha) treatment in leukemias of the B-cell lineage shows a marked heterogeneity. A distinct subset of patients with B-CLL responds to treatment with IFNalpha, while the drug has no therapeutic effect in the majority of patients. The mechanism of this phenomenon is poorly understood. The cellular events induced by this cytokine mediated by a number of specific signaling events. Therefore, we studied the effect of recombinant IFNalpha on tyrosine phosphorylation and proliferation of cytosolic proteins in human cell lines and in freshly isolated B-CLL cells in order to test the potential value of these events as a pretreatment test for IFNalpha in CLL. In human lymphoid cell lines, IFNalpha induced tyrosine phosphorylation of multiple cytosolic proteins in a time- and concentration-dependent manner. This effect correlated with its growth-inhibitory effect in almost all cell lines. In marked contrast, in freshly isolated B-CLL cells IFNalpha seemed to have both stimulatory and inhibitory effects on proliferation, but it consistently stimulated tyrosine phosphorylation. Moreover, the clinical response of B-CLL to IFNalpha did not correlate with the activation of tyrosine kinases nor with the inhibition of cell growth in vitro. Therefore, the assessment of IFNalpha-induced tyrosine phosphorylation of cytosolic phospho-proteins does not allow to predict the treatment response to IFNalpha in CLL patients. PMID- 10803934 TI - Expression of functional granulocyte colony-stimulating factor receptors on human B-lymphocytic leukemia cells. AB - We analyzed the expression of cell surface antigens and granulocyte colony stimulating factor (G-CSF) receptors using flow cytometry, the expression of G CSF mRNA receptor, using reverse transcription (RT)-PCR, and tested the effect of G-CSF on leukemia colony formation. A total of 14 lymphocytic leukemia patients were examined, seven with acute lymphocytic leukemia (ALL), two with adult T-cell leukemia (ATL), two with B-chronic lymphocytic leukemia (CLL), two with chronic myelocytic leukemia in lymphoid blastic crisis (CML-LBC), and one with plasma cell leukemia (PCL). The presence of G-CSF receptors was demonstrated in 4/14 (29%) patients, two with ALL, one with CLL, and one with CML-LBC, and was associated with stimulation of leukemia clonogenic cell growth by G-CSF. In addition, all four positive leukemia cell types expressed typical B-cell antigens. Our results indicated that G-CSF receptors are expressed on some portion of B-lymphoid leukemia and that their receptors are functional as growth stimulators. PMID- 10803935 TI - Palliative cytoreduction in refractory acute leukemia: a retrospective study of 57 adult patients. AB - The efficiency and toxicity of treatment regimens for nonintensive cytoreduction in 57 outpatients with refractory acute leukemia (mean age 56 years, 51 AML, six ALL/AUL) were retrospectively studied. Seventeen patients received one treatment regimen, 19 patients two treatment regimens, and 21 patients three or more treatment regimens. The treatment regimens analyzed were 6-thioguanine p.o. (daily) (T), 6-thioguanine p.o. (4-7 days/week) + cytarabine s.c./i.v. (once a week) (T+C), 6-mercaptopurine p.o. (daily) (MP), 6-mercaptopurine p.o. (daily) + methotrexate p.o./i.v. (once a week) (MP+MTX), etoposide p.o. (daily) (E), and mitoxantrone i.v. (M). The median leukocyte count was higher for M (73 x 10(9)/l) than for the other treatment regimens (T: 27 x 10(9)/l, T+ C: 37 x 10(9)/l, MP: 24 x 10(9)/l, MP + MTX: 30 x 10(9)/l, E: 31 x 10(9)/l). A cytoreduction >50% in the peripheral blood was achieved by T in 11/19, by T+C in 7/11, by MP in 5/8, by MP+MTX in 3/6, by E in 3/4, and by M in 16/22 patients. The period of cytoreduction was regarded as the duration of response - T: median 53 days, range 5-98; T+C: median 61 days, range 14-226; MP: median 37 days, range 4-192; MP + MTX: median 58 days, range 36-59; E: median 121 days, range 26-159; M: median 39 days, range 8-78. T and T + C were well tolerated by all but three patients (stomatitis, diarrhea, WHO grade 2). MP was accompanied by a rise of transaminases (WHO 1-3) in 5/6 patients. E led to stomatitis (WHO 1,2) in 4/5 and M to nausea/vomiting (WHO 1,2) in 5/22 and to stomatitis (WHO 2) in 4/22 cases. The mean survival time after start of palliative cytoreduction was 16 weeks (2 65). In summary, 6-thioguanine +/- cytarabine was best tolerated with effective but in oral monotherapy - often protracted cytoreduction in 60% of patients. Mitoxantrone showed tolerable side effects and potent cytoreduction in 73% of patients even after ineffective palliative pretreatment. Palliative cytoreductive therapy does not reduce the quality of life and can prevent complications of significant leukocytosis in refractory acute leukemia. PMID- 10803936 TI - A phase-II trial of all trans retinoic acid and low-dose cytosine arabinoside for the treatment of high-risk myelodysplastic syndromes. AB - Twenty-two patients with high-risk myelodysplastic syndrome (HRMDS) were treated with a 10-day course of oral all trans retinoic acid (45 mg/m2) and s.c. low-dose cytosine arabinoside (LDARAc) given at the dose of 20 mg twice per day. The courses were repeated monthly until response or progression, in the case of response, the therapy was administered until relapse. Morphologic diagnoses were refractory anemia with excess blasts (RAEB) in nine, RAEB in transformation (RAEB t) in nine, and chronic myelomonocytic leukemia (CMMoL) in four patients; in all cases, bone-marrow blast infiltration was greater than 10% (median 20%, range 12 30%). When the international prognostic scoring system was applied, all the cases qualified as intermediate/high-risk categories. Nineteen patients were males and three were females; the median age was 69 years (range 25-90 years); three patients had previously been treated with conventional chemotherapy, and one of them had also undergone autologous bone-marrow transplantation. The criteria of response were defined as follows: (1) complete response: normalization of blood counts and bone-marrow blasts (<5%), and (2) partial response: decrease in bone marrow blast infiltration by 50%, and two of the following parameters - improvement in hemoglobin level by 1.5 g/dl or decrease by 50% in transfusional requirement, increase by 50% in absolute neutrophil count, and increase by 50% in platelet count. Overall, 7 (32%) of 22 patients achieved a response, with 5 (23%) being classified as complete responders and 2 (9%) as partial responders. Fifteen (68%) patients did not achieve any response, and 14 died of progressive disease or infectious disease. The overall median survival was 8 months (range 1-27 months), whereas the median survival of responders was 16 months (range 8-27 months); the median duration of response was 11 months (range 2-21 months). Moderate to severe hematological toxicity and infections were the most common side effects. In conclusion, it seems that the association of ATRA and LDARA-C may be effective in approximately 30% of HRMDS patients. Optimizing this approach might be pursued by selecting, on a biological basis, those cases more likely to respond or by incorporating other differentiating agents or growth factors. PMID- 10803937 TI - Perioperative treatment with filgrastim stimulates granulocyte function and reduces infectious complications after esophagectomy. AB - We investigated the effects of recombinant G-CSF (Filgrastim) on the function of neutrophils and the rate of infectious complications in an open-label, nonrandomized study of patients with esophageal cancer undergoing esophagectomy. In this single-center phase-II trial 20 sequential patients (19 evaluable) received Filgrastim at standard doses (300 microg or 480 microg) subcutaneously for 2 days prior to and up to 7 days after surgery. The phagocytotic activity of neutrophils and the oxidative burst in the study group and in an experimental control group (n=27) were measured on days -2, 2, and 10. Neutrophil function was enhanced in the Filgrastim-treated group by factor 1.2 for phagocytosis (p=0.016) and 1.4 for oxidative burst (p)=0.154). Leukocyte counts increased from 7.6 x 10(9)/l (day -2) to a maximum of 45 x 10(9)/l on day 6. No infection was reported in the study group (mean age 59.7 years; 13 men, seven women) up to 10 days after surgery. In contrast, 23 patients (29.9%) in a historical control group (mean age 56 years; 67 men, ten women) treated at the same center developed infections within the first 10 days (p = 0.008). In addition, no postoperative deaths occurred in the study group, compared with 9.1% in the group of historical controls. Thus, in this study, administration of Filgrastim stimulated neutrophil function in patients undergoing esophagectomy, and it might be effective in reducing infectious complications related to the surgical procedure. PMID- 10803938 TI - Meropenem versus ceftazidime as empirical monotherapy for febrile neutropenic cancer patients. AB - A total of 101 cancer patients with 121 febrile neutropenia episodes were randomised to receive empirical treatment with i.v. meropenem (1g/8 h) or ceftazidime (2 g/8 h). After 3 days, 89% of patients were on unmodified therapy in the meropenem group, compared with 83% in the ceftazidime group. Of the evaluable episodes (n = 106), the success rate with unmodified empirical therapy until the end of the treatment course was slightly higher with meropenem than with ceftazidime (48% vs 38%, P=0.39). Furthermore, initial success with further infections was observed in 22% of episodes treated with meropenem and in 13% of episodes treated with ceftazidime. Glycopeptides were used as first modification in 28% and 39% of meropenem and ceftazidime recipients, respectively. Both treatments were well tolerated and there were no reports of drug-related nausea/vomiting or seizures. No significant differences in response rate or in tolerability were observed when analysing only the first febrile episodes. In conclusion, meropenem seems to be as efficacious and well tolerated as ceftazidime and may be associated with a lesser requirement for the addition of glycopeptides. PMID- 10803939 TI - Fatal rupture of the spleen caused by infiltration of T-cell lymphoma. AB - Pathological or spontaneous rupture of the spleen has been described in a variety of diseases affecting the spleen, with infections being cited as the cause in most cases. In hematological malignancies it is a rare event, despite the frequent involvement of the spleen in these diseases. It has, however, been described in patients with acute and chronic leukemia, Hodgkin's disease, non Hodgkin's lymphoma of B-cell origin, mycosis fungoides, and so-called histiocytic lymphoma. Here, we present a fatal case of splenic rupture caused by infiltration of a peripheral T-cell lymphoma, unspecified according to the REAL classification. The importance of a correct diagnosis and fast surgery is emphasized. PMID- 10803940 TI - Sudden onset of EDTA-dependent pseudothrombocytopenia after therapy with the glycoprotein IIb/IIIa antagonist c7E3 Fab. AB - The development of thrombocytopenia following exposure to the platelet glycoprotein (GP) IIb/ IIIa receptor antagonist abciximab (c7E3 Fab, ReoPro) is associated with adverse clinical outcome and excessive bleeding. Pseudothrombocytopenia is an important differential diagnosis in sudden onset of thrombocytopenia in a patient treated with c7E3 Fab. We report on a case of documented sudden onset of EDTA-dependent pseudothrombocytopenia in a 63-year-old woman who was admitted for emergency coronary intervention. Four hours after bolus administration of c7E3 Fab, the platelet concentration in EDTA anticoagulated blood decreased from 385 x 10(9)/l to 119 x 10(9)/l, and it showed a further decrease to 57 x 10(9)/l at the end of a 12-h infusion. Despite no warnings or abnormalities of the automated cell counter, platelet aggregates were observed by microscopic evaluation of the blood smear. Repeated platelet counts in citrate-anticoagulated samples revealed platelet concentrations within the reference range. EDTA-dependent pseudothrombocytopenia due to therapy with c7E3 Fab is an important differential diagnosis that needs to be excluded rapidly from other causes of thrombocytopenia to avoid unnecessary further examinations, discontinuation of therapy, or even initiation of inappropriate therapy. PMID- 10803941 TI - Enhancement of plant stem growth by flocculation of the antibiotic-producing bacterium, Pseudomonas fluorescens S272, on the roots. AB - The antibiotic-producing bacterium, Pseudomonas fluorescens, is assumed to be important in protecting plants from soilborne diseases. S. fluorescens S272, a hyper-producing strain of pyoluteorin (PT) and 2,4-diacetylphloroglucinol (DG), had previously been isolated from soil. The present paper reported that the growth of water-cultivated Kaiware radish was promoted to 120-140% of its normal level by the coaddition of an S272 culture broth (0.01-1% v/v) and a polysaccharide flocculant (1-100 ppm) from Klebsiella pneumoniae H12. Tight adhesion of S272 cells to the root tissue was microscopically observed. The growth promotion is assumed to have been caused by antibiotic effects for the following two reasons: 1) PT (4 mg/l) and DG (24 mg/l) addition to a radish culture enhanced stem growth to 130% of the normal level; 2) a culture solution containing the S272 culture broth (0.01-1% v/v) markedly inhibited the decomposition of hypersensitive chrysanthemum leaves. A soil-cultivation experiment with Gomphrena globosa under natural conditions also exhibited enhanced stem length (160%) by coaddition of the S272 culture broth and H12 polysaccharide. These results suggest that polysaccharide-enhanced adhesion of P. fluorescens S272 cells might be useful for promoting plant growth through the increased antibiotic effect. PMID- 10803943 TI - Interconversion between dehydro-L-ascorbic acid and L-ascorbic acid. AB - L-Ascorbic acid (AA) plays an important role in biological systems as an electron donor. Erythorbic acid (EA) is the epimer of AA and has chemical characteristics very similar to those of AA. It is demonstrated in the present study by 1H-NMR that dehydro-L-ascorbic acid (DAA) was reduced by EA under neutral conditions but not acidic, and that dehydroerythorbic acid (DEA) was also reduced by AA under the same conditions. These reactions also occurred at a low concentration close to the concentration of AA in such biological tissue as the liver. Furthermore, the interconversion of DAA and AA at neutral pH and low concentration was also confirmed by radioluminography. These results suggest the interconversion between DAA and AA in vivo. PMID- 10803942 TI - Lower plasma triglyceride level in Syrian hamsters fed on skim milk fermented with Lactobacillus casei strain Shirota. AB - The effect of fermented skim milk (FSM) by Lactobacillus casei strain Shirota on plasma lipids in hamsters was examined. Hamsters fed on cholesterol-free and enriched diets containing 30% FSM had lower levels of plasma triglyceride than those fed on the control diet. In the experiment with the cholesterol-enriched diet-fed hamsters, the plasma triglyceride level was suppressed by FSM at concentrations of 10% to 30%. Unfermented milk tended to lower the level of triglyceride, but not significantly. The plasma cholesterol concentration was not affected by an FSM and unfermented skim milk supplement to the diet. L. casei strain Shirota grew well in the presence of mixed lipid micelles containing bile acid, but did not have the ability to remove cholesterol from the culture broth. These results indicate that FSM lowered the plasma triglyceride level in hamsters. PMID- 10803944 TI - Role of the DNA sequence downstream of the Bacillus subtilis hut promoter in regulation of the hut operon. AB - To identify the role of the downstream region of a hut promoter in regulation of the Bacillus subtilis hut operon, three single-base substitutions (+9G-->A, +14C- >T, and +23T-->G) were introduced into the hut operon. Analysis of expression of the hut operon containing each of these three single-base substitutions and the hut-lacZ fusions with the single-base substitutions at position +14 showed that the position at +14 and probably the position at +23 were required for amino acid repression at the hut promoter, while the position at +14 was not required for catabolite repression at the hut promoter. The position at +9 was required for a histidine-dependent increase of activity of the hut promoter. Analysis of expression of the hut-lacZ fusions and the hut operon in the codY mutant indicated that the position at +14 and probably the position at +23 were involved in CodY-mediated amino acid repression at the hut promoter and that CodY was not required for catabolite repression at the hut promoter. PMID- 10803945 TI - Purification and characterization of membrane-bound hydrogenase from Hydrogenobacter thermophilus strain TK-6, an obligately autotrophic, thermophilic, hydrogen-oxidizing bacterium. AB - A membrane-bound hydrogenase was purified to electrophoretic homogeneity from the cells of Hydrogenobacter thermophilus strain TK-6, an obligately autotrophic, thermophilic, hydrogen-oxidizing bacterium. Solubilization and purification were done aerobically in the presence of Triton X-100. Three chromatography steps were done for purification; Butyl-Sepharose, Mono-Q, and Superose 6, in this order. Purification was completed with 6.73% yield of total activity and with 21.4-fold increase of specific activity when compared with the values for the membrane fraction. The purified hydrogenase was shown to be a tetramer with alpha2beta2 structure, with a molecular mass of 60,000 Da for the large subunit and 38,000 Da for the small subunit. The purified hydrogenase directly reduced methionaquinone with an apparent Km of around 300 microM and with a turnover number around 2900 (min(-1)). Metal analysis and EPR properties of the hydrogenase have shown that the enzyme is one of the [NiFe]-hydrogenases. Also, optimum pH and temperature for reaction, thermal stability, and electron acceptor specificity were reported. Finally, a model is presented for energy and central metabolism of H. thermophilus strain TK-6. PMID- 10803946 TI - Inhibition of radiation-induced lipid peroxidation by tetrahydrocurcumin: possible mechanisms by pulse radiolysis. AB - The antioxidant property of tetrahydrocurcumin (THC), a reduced derivative of curcumin, was examined by its ability to inhibit radiation-induced lipid peroxidation in rat liver microsomes and compared with curcumin. The lipid peroxidation caused by irradiation of N2O-purged and aerated buffered aqueous solutions was found to be inhibited by THC in a dose- and concentration-dependent manner. In order to understand the actual reaction mechanisms involved in the inhibition process, pulse radiolysis investigation of THC with radiolytically produced radicals like hydroxyl, model peroxyl radicals, and azide radicals were done and the transients were detected by kinetic spectrophotometry. The reaction of THC with hydroxyl and azide radicals gave rise to transient absorption in the region 200-400 nm with two peaks at 310 nm and 390 nm. From the spectral properties and kinetics of these radicals, a suitable mechanism is discussed to explain the antioxidant actions of THC. PMID- 10803947 TI - A simple and rapid method for the detection of poly(ADP-ribose) by flow cytometry. AB - Measurement of poly(ADP-ribose) levels was performed by a new method using a monoclonal antibody against poly(ADP-ribose) and flow cytometry from small amount of cultured cells without the need for isolation of poly(ADP-ribose) polymer. The increase of poly(ADP-ribose)-associated fluorescence intensity was observed in individual human leukemic HL-60 cells when treated with the carcinogen, N-methyl N'-nitro-N-nitrosoguanidine (MNNG), and was blocked by the treatment with 3 aminobenzamide before MNNG treatment. It is easy and rapid to detect the time dependent change of poly(ADP-ribose) levels in HL-60 cells after MNNG treatment. We easily found that the increase of the poly(ADP-ribose) level in nicotinic acid treated lymphocytes after MNNG treatment was observed, but not in nicotinamide treated lymphocytes. We investigated the change of poly(ADP-ribose) levels especially in the early phase of apoptosis. Our method is simple and rapid. It is suggested that the investigation of poly(ADP-ribosyl)ation in various fields is possible by using this new method. PMID- 10803948 TI - Detection of the cholera toxin-binding activity of kappa-casein macropeptide and optimization of its production by the response surface methodology. AB - The cholera toxin (CT)-binding activity of purified kappa-casein macropeptide (CMP) from bovine kappa-casein was detected. In addition, a statistical model was developed to optimize the production of CMP. CMP was prepared by chymosin hydrolysis of kappa-casein and a subsequent 3% trichloroacetic acid treatment. CMP was further fractionated in an ion-exchange column by FPLC. CT binding activity was eluted at 0.18 M NaCl and was a single 8.9 kDa peptide without tyrosine and arginine residues. The CT binding activity was rapidly lost by a carbohydrase treatment. The conditions for CMP production with chymosin were optimized by using the response surface methodology (RSM). The estimated optimum levels of the factors were as follows: reaction temperature, 38.5 degrees C; pH, 6.44; and time, 35.9 min. A validation experiment was performed in which CMP was prepared under the predicted parameters, and it was ascertained that the estimated optimum conditions gave better production of CMP than any other conditions. PMID- 10803949 TI - Cloning, sequencing, and expression of UDP-glucose pyrophosphorylase gene from Acetobacter xylinum BRC5. AB - A UDP-glucose pyrophosphorylase (UGPase) gene from Acetobacter xylinum BRC5 has been cloned, sequenced, and expressed in Escherichia coli. The gene consists of 867 nucleotides and encodes a polypeptide of 289 amino acid residues with a calculated molecular mass of 31,493 Da. The amino acid sequences of the enzyme showed an 85.8% identity to those of an enzyme from A. xilinum ATCC 23768. A polyhistidine-UGPase fusion enzyme was expressed and purified from the transformed E. coli. The enzyme showed a 35,620-Da single protein band on SDS/PAGE and an about 160,000-Da protein band on 8-16% pore-gradient polyacrylamide gel, indicating the enzyme may be a tetramer or pentamer composed of four or five identical subunits. Kinetic analysis of the enzyme showed a typical Michaelis-Menten substrate saturation pattern, from which Km and Vmax were calculated to be 3.22 mM and 175.4 micromol x min(-1) x mg(-1) for UDP glucose and 0.24 mM and 69.4 micromol x min(-1) x mg(-1) for PPi, respectively, required Mg2+ for maximal activity, and was inhibited by free pyrophosphate. Computer-aided comparison of the Acetobacter enzyme sequence with those of other bacterial enzymes found significant similarities among them and predicted that Lys84 is a catalytically important residue. Lys84 in the enzyme, which was also conserved in other bacterial enzyme sequences, was replaced by arginine or leucine. The K84R mutant enzyme was successfully expressed in E. coli and showed enzyme activity (63% of the wild-type enzyme activity), but K84L was not isolated in stable form. These results suggest that Lys84 is significant in not only catalysis but also maintenance of the active structure. PMID- 10803950 TI - Substrate diversity of macrophomate synthase catalyzing an unusual multistep transformation from 2-pyrones to benzoates. AB - Macrophomate synthase, which we have recently purified, catalyzes an unusual multistep transformation from 5-acetyl-4-methoxy-6-methyl-2-pyrone to 4-acetyl-3 methoxy-5-methyl-benzoic acid (macrophomic acid). To investigate the substrate diversity of the enzyme, 40 analogs of 2-pyrone were prepared and their relative efficiency was examined in the enzymatic conversions. The experimental results reveal the structural requirements of the substrates and the rough size of the enzyme active site, and eliminate the ambiguity caused by contamination by other enzymes in the whole-cell experiments. PMID- 10803951 TI - Biosynthesis of violacein: intact incorporation of the tryptophan molecule on the oxindole side, with intramolecular rearrangement of the indole ring on the 5 hydroxyindole side. AB - Feeding experiments with a mixture of [2-13C]- and [indole-3-13C]tryptophans, of [3-13C]- and [indole-3-13 C]tryptophans (1:1 molar ratio) and of others have proved that the 1,2-shift of the indole ring occurred via an intramolecular process for formation of the left part (5-hydroxyindole side) of the violacein skeleton and demonstrated that the C-C bond from C2 of the indole ring to C2 of the side chain was completely retained for formation of the right part (oxindole side) during the entire biosynthetic process. Due to the involvement of transaminase, it has remained unresolved whether indolylpyruvic acid is the biosynthetic intermediate and/or from where the nitrogen atom of the pyrrolidone ring originates. An incorporation experiment with a mixture of [2-13C]- and [alpha-15N]tryptophans (1:1 molar ratio) verified that the nitrogen atom in the central ring was exclusively derived from the right-side tryptophan. Thus, all the carbon and nitrogen atoms in the right part of the violacein skeleton were constructed by intact incorporation of the tryptophan molecule, with decarboxylation probably occurring at a later biosynthetic stage. PMID- 10803952 TI - Characteristics of the biotin enhancement of glucose-induced insulin release in pancreatic islets of the rat. AB - Perifused isolated rat islets were used to show that biotin plus 16.5 mM glucose evoked more insulin secretion than 16.5 mM glucose alone. Whether or not this reinforcement of glucose-induced insulin secretion by biotin is unique was studied by using perifused islets stimulated with 16.5 mM glucose plus 100 microM of one of various components of the vitamin B group. No effect of any of these vitamins was found on glucose-induced insulin secretion. These results indicate that biotin is unique among the members of the vitamin B group in enhancing glucose-induced insulin secretion. Static incubation experiments showed that biotin did not potentiate insulin release when the islets were incubated with an experimental solution containing either no or 2.8 mM glucose. The addition of biotin to 27.7 mM glucose, which is the maximal concentration for stimulating insulin release, did not significantly enhance the effect of the glucose on insulin release (although it did at 16.5 mM glucose). These findings indicate that biotin, by itself, does not stimulate insulin secretion, and does not enhance glucose-induced insulin secretion beyond the ability of glucose itself to stimulate insulin secretion. PMID- 10803953 TI - Increased hepatic activity of inducible nitric oxide synthase in rats fed on a high-fat diet. AB - The effects were examined of the dietary level of fat on the activity of inducible nitric oxide synthase (iNOS) in the liver of rats. In experiment 1, rats were fed on a diet containing 5% or 20% beef tallow or safflower oil for 32 d. The animals were given a subcutaneous injection of the carcinogen, 1,2 dimethylhydrazine (DMH), on d 4. The activity of hepatic iNOS was significantly elevated by the high-fat diet, but was unaffected by the dietary source of the fat examined. In experiment 2, rats were fed on a 5% or 20% beef tallow diet for 11 d or 32 d with or without the DMH treatment. Feeding the high-fat diet and DMH treatment caused higher activity of hepatic iNOS. In experiment 3, the high-fat diet elevated hepatic iNOS activity and the amount of its protein in the lipopolysaccharide-treated rats. The results suggest that hepatic NO production is enhanced by a high-fat diet. PMID- 10803954 TI - Changes in N-linked oligosaccharides during seed development of Ginkgo biloba. AB - Structural changes in N-linked oligosaccharides of glycoproteins during seed development of Ginkgo biloba have been explored to discover possible endogenous substrate(s) for the Ginko endo-beta-N-acetylglucosaminidase (endo-GB; Kimura, Y., et al. (1998) Biosci. Biotechnol. Biochem., 62, 253-261), which should be involved in the production of high-mannose type free N-glycans. The structural analysis of the pyridylaminated oligosaccharides with a 2D sugar chain map, by ESI-MS/MS spectroscopy, showed that all N-glycans expressed on glycoproteins through the developmental stage of the Ginkgo seeds have the xylose-containing type (GlcNAc2 approximately 0Man3Xyl1Fuc1 approximately 0GlcNAc2) but no high mannose type structure. Man3Xyl1Fuc1GlcNAc2, a typical plant complex type structure especially found in vacuolar glycoproteins, was a dominant structure through the seed development, while the amount of expression of GlcNAc2Man3Xyl1Fuc1GlcNAc2 and GlcNAc1Man3Xyl1Fuc1GlcNAc2 decreased as the seeds developed. The dominantly occurrence of xylose-containing type structures and the absence of the high-mannose type structures on Ginkgo glycoproteins were also shown by lectin-blotting and immunoblotting of SDS-soluble glycoproteins extracted from the developing seeds at various developmental stages. Concerning the endogenous substrates for plant endo-beta-N-acetylglucosaminidase, these results suggested that the endogenous substrates might be the dolicol oligosaccharide intermediates or some glycopeptides with the high-mannose type N glycan(s) derived from misfolded glycoproteins in the quality control system for newly synthesized glycoproteins. PMID- 10803955 TI - Molecular analysis of maleate cis-trans isomerase from thermophilic bacteria. AB - Several strains of thermophilic bacteria containing maleate cis-trans isomerase were isolated from soil samples and identified as Bacillus stearothermophilus, Bacillus circulans, Bacillus brevis, and Deleya halophila. The maleate cis-trans isomerase was purified and characterized from one of the isolated strains, B. stearothermophilus MI-102. The purified enzyme of strain MI-102 showed higher thermal stability than the enzyme of a mesophile, Alcaligenes faecalis IFO13111. The seven maleate cis-trans isomerase genes (maiA) of thermophile were cloned and sequenced. B. stearothemophilus MI-102 MaiA has 67% amino acid identity with A. faecalis MaiA. All eight amino acid sequences of maiA gene products had significant conserved regions containing cysteine residues, which were previously suggested to be involved in an active site of the enzyme. To probe the catalytic mechanism, three cysteine residues in the conserved regions of A. faecalis MaiA were replaced with serine by site-directed mutagenesis. The results suggest that Cys80 and Cys198 play important roles in the enzyme activity. PMID- 10803956 TI - Methionine-induced phytoalexin production in rice leaves. AB - The application of methionine on wounded rice leaves induced the production of rice phytoalexins, sakuranetin and momilactone A. This induction resulted from stimulation of phenylalanine ammonia-lyase and naringenin 7-O-methyltransferase activity. Jasmonic acid, ethylene, and active oxygen species are important as signal transducers in disease resistance mechanisms. However, although the endogenous level of jasmonic acid rapidly increased in reaction to wound, methionine treatment could not induced endogenous JA production. Ethylene induced the production of the flavonoid phytoalexin, sakuranetin, but did not induce the production of a terpenoid phytoalexin, momilactone A. On the other hand, a free radical scavenger, Tiron, counteracted the induction of both sakuranetin and momilactone A production in methionine-treated leaves. Active oxygen species may be important in methionine-induced production of phytoalexins. PMID- 10803957 TI - Glycosidic fraction of flue-cured tobacco leaves: its separation and component analysis. AB - The fraction containing glycosidic components was separated from flue-cured tobacco (Nicotiana tabacum L.) leaves by a facile method. Some components of the fraction were isolated and elucidated to be syringin, coniferin, cichoriin, benzyl-beta-D-glucoside, Blumenol A-beta-D-glucoside, and 5,6-epoxy-5,6-dihydro-3 hydroxy-beta-ionyl-beta-D-glucoside. Syringin and coniferin were detected in the Nicotiana species for the first time. PMID- 10803958 TI - Suppressive effect of excess dietary histidine on the expression of hepatic metallothionein-1 in rats. AB - The gene expression of liver metallothionein-1 in excess dietary histidine was investigated by feeding rats ad libitum on either a basal or histidine-excess (50 g of L-histidine per kg of diet) diet for 5 d. The copper content of the liver and zinc level in the serum of the rats fed on the histidine-excess diet were lower by 21% and 61%, respectively of the figures for the rats fed on the basal diet, but the zinc content of the liver and copper level in the serum were not affected. Excess dietary histidine caused an increase in the urinary output of both copper and zinc. The level of liver metallothionine-1 mRNA was markedly lower at 19% in the rats fed on the a histidine-excess diet compared to the level with the basal diet. It thus appears that such a response by the level of liver metallothionein-1 mRNA might have been be due to the lower content of liver copper. PMID- 10803959 TI - Detection of a spontaneous high expression of heat shock protein 70 in developing zebrafish (Danio rerio). AB - A spontaneous high expression of heat shock protein 70 (HSP 70) was detected in zebrafish (Danio rerio) at early larval stage (84 h after fertilization), but the HSP 70 level was either low or barely detectable in 12, 24, 36, 60, and 108 h after fertilization. The extracts of zebrafish at 80 and 84 h after fertilization formed a clear protein-DNA complex with a probe containing heat shock elements (HSEs), suggesting that this spontaneous expression of HSP 70 may be turned on via the binding of stage-specific HSE-binding factors to HSP 70 gene promotor. The protein-HSE complexes produced by the spontaneous binding, however, were found to be different from those formed by the extracts of heat-treated zebrafish in electrophoretic mobility. PMID- 10803960 TI - Alkylated benzothiophene desulfurization by Rhodococcus sp. strain T09. AB - A benzothiophene desulfurizing bacterium was isolated and identified as Rhodococcus sp. strain T09. Growth assays revealed that this strain assimilated, as the sole sulfur source, various organosulfur compounds that cannot be assimilated by the well-studied dibenzothiophene-desulfurizing Rhodococcus sp. IGTS8. The cellular growth rate of strain T09 for the alkylated benzothiophenes depended on the alkylated position and the length of the alkyl moiety. PMID- 10803961 TI - Preparation of (-)-periplanone D and its physical and spectroscopic properties. AB - (-)-Periplanones C and D were obtained in reproducible yields by modifying reported procedures. Our synthetic sample of (-)-periplanone D showed somewhat different physical and spectroscopic properties from those reported in the literature. PMID- 10803962 TI - Expression and mutational analysis of amino acid residues involved in catalytic activity in a ribonuclease MC1 from the seeds of bitter gourd. AB - The ribonuclease MC1 (RNase MC1) from seeds of bitter gourd (Momordica charantia) consists of 190 amino acids and belongs to the RNase T2 family, including fungal RNases typified by RNase Rh from Rhizopus niveus. We expressed RNase MC1 in Escherichia coli cells and made use of site-directed mutagenesis to identify essential amino acid residues for catalytic activity. Mutations of His34 and His88 to Ala completely abolished the enzymatic activity, and considerable decreases in the enzymatic activity were observed in cases of mutations of His83, Glu84, and Lys87, when yeast RNA was used as a substrate. Kinetic parameters for the enzymatic activity of the mutants of His83, Glu84, and Lys87 were analyzed using a dinucleoside monophosphate CpU. Km values for the mutants were approximately like that for wild-type, while k(cat) values were decreased by about 6 to 25-fold. These results suggest that His34, His83, Glu84, Lys87, and His88 in RNase MC1 may be involved in the catalytic function. These observation suggests that RNase MC1 from a plant catalyzes RNA degradation in a similar manner to that of fungal RNases. PMID- 10803963 TI - Group I intron located in PR protein homologue gene in Youngia japonica. AB - A Youngia japonica strain had a group I intron that was suggested to have been transferred from Protomyces inouyei, a pathogenic fungus of Y. japonica. It was located in the miraculin homologue coding gene by reverse complementation. The deduced amino acid sequence of this miraculin homologue of Y. japonica was similar to the amino acid sequences of tobacco and tomato pathogenesis-related proteins. PMID- 10803964 TI - Synthesis of 2,2-diphenylpropionate derivatives and their effects on larval growth of the silkworm. AB - A variety of 2,2-diphenylpropionate derivatives with an amino substituent were synthesized and their effects on larval growth of the silkworm, Bombyx mori, were examined by dietary administration. Of the compounds tested, 3-(4-ethylpiperazin 1-yl)propyl 2,2-diphenylpropionate (3) caused significant prolongation of the larval period. Studies on the structure-activity relationship indicate that a piperazine ring and the bond distances between the nitrogen atom and the ester group were important for this activity. Treatment of compound 3 delayed the increase of ecdysteroid titers in the hemolymph by 3 days compared with that of the control, which correlates with the delay in molting. PMID- 10803965 TI - Increased intestinal calcium absorption from the ingestion of a phosphorylated guar gum hydrolysate independent of cecal fermentation in rats. AB - The Apparent calcium absorption was increased in rats fed on P-GGH and GGH. However, this increase in calcium absorption from GGH feeding was cancelled by a cecectomy, whereas the corresponding increase from P-GGH feeding was not. The change in femoral calcium content was similar to that in calcium absorption. The calcium solubility in the ileum was increased in those rats fed on P-GGH. We conclude that cecal fermentation did not contribute to the increased calcium absorption by the rats fed on P-GGH. PMID- 10803966 TI - Changes in gamma-aminobutyric acid content during beni-koji making. AB - The changes in the gamma-aminobutyric acid (GABA) content during the making of beni-koji prepared with Monascus pilosus IFO 4520 vs. the difference in the rate of tomo koji (10%, 30%, and 50%) were examined. The increased proportion of tomo koji would increase the GABA production and the productions of GABA peaked on the fifth day and thereafter declined. The glutamate decarboxylase activity during beni-koji making with 50% tomo koji steadily increased after the start of the koji making, reaching its peak on the fifth day. The succinic acid content increased after the sixth day. The mycelial growth was in the stationary phase after the sixth day. Therefore, the GABA content increases with an increase in the proportion of tomo koji. It is presumed that the maximum amount of GABA reached on the fifth day was the cause of the increasing amount of conversion of GABA into succinic acid, in addition to the decline in the GAD activity after the fifth day of koji making. PMID- 10803967 TI - Novel fungal metabolites, demethylsorbicillin and oxosorbicillinol, isolated from Trichoderma sp. USF-2690. AB - The novel compounds, demethylsorbicillin (1) and oxosorbicillinol (2), were isolated from a fermentation broth of Trichoderma sp. USF-2690. The structures of these compounds, which were determined from spectroscopic evidence, suggest the possibility that methylation at C-6 and oxidation at C-1 and C-6 of sorbicillin were controlled in the early polyketide stage before the formation of oxidized sorbicillin dimers. In a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay, 2 gave an ED50 value of 87.7 microM. PMID- 10803968 TI - Isolation and some properties of a serine protease from the fruits of Cudrania cochinchinensis (Lour.) Kudo et Masam. AB - An endopeptidase (Cudrania protease) with a molecular mass of 76 kDa has been purified from the fruits of Cudrania cochinchinensis (Lour.) Kudo et Masam. The enzyme was stable between pH 6 and 10 at 30 degrees C for 60 min. The enzyme activity was inhibited by diisopropyl fluorophosphate, chymostatin, and aprotinin, but not by EDTA or pepstatin. These results indicated that the enzyme was a serine protease. PMID- 10803969 TI - Absolute structure of N-p-coumaroyloctopamine in elicitor-treated potato tuber tissue. AB - Treatment of potato tuber tissue with beta-1,3-oligoglucosaccharide causes an accumulation of N-p-coumaroyloctopamine (1). In order to determine the absolute structure of 1 in potato, optically active 1 was synthesized from (R)-octopamine which had been obtained from the racemic mixture by the fractional crystallization. By comparing the chromatographic behavior of synthetic and naturally-occurring samples with a chiral HPLC analysis, the absolute configuration of 1 in potato was determined to be S. This indicates that the absolute configuration of the octopamine moiety of 1 is opposite to that of octopamine formed in animal tissues. PMID- 10803970 TI - Characterization and amino acid sequences of cytochromes c6 from two strains of the green alga Chlorella vulgaris. AB - Cytochromes c6 from the green algae Chlorella vulgaris CK-5 (CK5cyc6) and C. vulgaris CK-22 (CK22cyc6) were characterized and their amino acid sequences were analyzed. CK5cyc6 had a molecular mass of 9.3 kDa, isoelectric points of 3.0 (reduced) and 3.6 (oxidized), and a redox potential of +362 mV at pH 7.0. CK22cyc6 had a molecular mass of 9.5 kDa, isoelectric points of 2.9 (reduced) and 3.5 (oxidized), and a redox potential of +355 mV at pH 7.0. The absorption spectra of both cytochromes c6 showed 4 maxima in reduced form, and 2 maxima and a weak peak at 695 nm in oxidized form. The pyridine ferrohemochrome spectra indicated that their prosthetic group was heme c. These physicochemical properties were similar to those of other algal cytochromes c6. The amino acids (88 residues) of CK5cyc6 and CK22cyc6 were sequenced and the sequence motif CXXCH-, which is typical of the heme-binding site of c-type cytochrome, was clearly confirmed in both cytochromes. Twenty-six amino acid residues were substituted, and the similarity score of each of them was 70.45%. PMID- 10803971 TI - Purification of membrane-bound lactoferrin from the human milk fat globule membrane. AB - Although lactoferrin is known as a basic soluble glycoprotein, the presence of the membrane-bound form of this protein has also been demonstrated in human milk. Membrane-bound lactoferrin was extracted from the human milk fat globule membrane with a detergent mixture of 1% Tween-20, 0.5% C12E8, and 0.5 M KCl in 20 mM Tris HCl (pH 7.4). Lactoferrin in the detergent-soluble fraction was purified by affinity chromatography with Concanavalin A and by hydrophobic chromatography with phenyl-Superose. The purified protein gave a single band of 80 kDa by SDS PAGE. Its N-terminal amino acid sequence was consistent with that of human lactoferrin. PMID- 10803972 TI - A homologue of EGL1 encoding endo-1,4-beta-glucanase in elongating pea stems. AB - A gene (EGL2) encoding an endo-1,4-beta-glucanase in peas has been cloned as a homologue of EGL1. EGL2 encodes a polypeptide of 506 amino acids, including a 24 mer putative signal polypeptide. The gene product contains a domain conserved in endo-1,4-beta-glucanase (family 9) showing 60% amino acid identity to EGL1. EGL2 mRNA was accumulated only in the elongating regions of pea stems, although EGL1 mRNA was abundant in both elongating and non-elongating tissues. However, the level of EGL2 mRNA was not increased by the treatment with sucrose and auxin in pea segments. These results suggest that the expression of EGL2 either requires the presence of other factors related to the auxin effect or occurs independent of auxin in the elongating pea stems. PMID- 10803973 TI - Molecular cloning and characterization of rpbA encoding RNA polymerase II largest subunit from a filamentous fungus, Aspergillus oryzae. AB - We have cloned rpbA encoding the RNA polymerase II largest subunit (polIIL) from a filamentous fungus, Aspergillus oryzae. The rpbA product included eight highly conserved regions and the carboxyl-terminal domain (CTD). A. oryzae polIIL CTD with 184 amino acids was composed of 25 CTD consensus repeats, which was a similar number to those of lower eukaryotes. The amino acids in each repeat of A. oryzae polIIL, however, conformed less to the CTD consensus than those of polIILs from other lower eukaryotes. PMID- 10803974 TI - Synthesis of 2,2,4,4-tetramethyl-N,N'-bis(2,6-dimethylphenyl)cyclobutane-1,3 diimine , a unique compound from Arundo donax, and its analogues to test their antifeedant activity against the boll weevil, Anthonomus grandis. AB - 2,2,4,4-Tetramethyl-N,N'-bis(2,6-dimethylphenyl) cyclobutane-1,3-diimine (1), which was isolated from the Thai plant Arundo donax as an antifeedant against the boll weevil (Anthonomus grandis), and its analogues (9-13) were synthesized and shown to possess no remarkable antifeedant activity of practical interest. PMID- 10803975 TI - Establishment of monoclonal antibodies specific for Bacillus subtilis DB9011. AB - Bacillus subtilis DB9011 is a strain with useful functions for agriculture. To establish a method for the discrimination of this strain from others, monoclonal antibodies (MAbs) were prepared. Although two established MAbs (MAb9B6 and MAb14D2) cross-react with some other Bacillus strains in ELISA, only B. subtilis DB9011 vegetative cells are recognized by both MAbs. MAb14D2 recognizes flagellin, a 34-kDa unit protein of flagella. The two MAbs established will provide powerful tools with which detailed analysis of this bacterial strain can be obtained under environmental conditions. PMID- 10803976 TI - GTP, an inhibitor of transglutaminases, is hydrolyzed by tissue-type transglutaminase (TGase 2) but not by epidermal-type transglutaminase (TGase 3). AB - Epidermal-type transglutaminase (TGase 3) is devoid of GTPase activity, but its TGase activity is inhibited by GTP as in the case of tissue-type TGase (TGase 2). In addition, the inhibition was not affected by the presence of higher concentrations of Ca ion. These results indicate that GTP interacts with TGase 3 in a manner different from its action on TGase 2. PMID- 10803977 TI - Cloning of a full-length cDNA encoding ent-kaurene synthase from Gibberella fujikuroi: functional analysis of a bifunctional diterpene cyclase. AB - We report here the nucleotide sequence of a full-length cDNA encoding ent-kaurene synthase that was isolated by a reverse-transcription polymerase chain reaction from Gibberella fujikuroi (Gcps/ks). This cDNA encodes 952 amino acid residues with a relative molecular mass of 107 kDa. The sequence similarity between Gcps/ks and ent-kaurene synthase of the gibberellin A1-producing fungus, Phaeosphaeria sp. L487, is very high, suggesting that Gcps/ks is also a bifunctional diterpene cyclase. Its recombinant protein expressed in Escherichia coli converted geranylgeranyl diphosphate to copalyl diphosphate and ent-kaurene. PMID- 10803978 TI - Efficient production of N-terminally truncated biologically active human interleukin-6 by Bacillus brevis. AB - cDNAs encoding human interleukin 6 (hIL-6) and its variants lacking the N terminal Pro and Pro-Val-Pro-Pro, respectively, were expressed in Bacillus brevis by using the signal peptide fusion approach. The presence of Pro at the N terminus of the mature protein hindered the action of the Bacillus brevis signal peptidase. hIL-6 lacking the N-terminal Pro-Val-Pro-Pro was most efficiently secreted in a biologically active form and accumulated in the culture medium to a level of 200 mg per liter, which is the highest level reported for the bacterial secretion of hIL-6. PMID- 10803979 TI - An automated system for genome analysis to support microbial whole-genome shotgun sequencing. AB - We developed a semi-automated genome analysis system called GAMBLER in order to support the current whole-genome sequencing project focusing on alkaliphilic Bacillus halodurans C-125. GAMBLER was designed to reduce the human intervention required and to reduce the complications in annotating thousands of ORFs in the microbial genome. GAMBLER automates three major routines: analyzing assembly results provided by genome assembler software, assigning ORFs, and homology searching. GAMBLER is equipped with an interface for convenience of annotation. All processes and options are manipulatable through a WWW browser that enables scientists to share their genome analysis results without choosing computer platforms. PMID- 10803980 TI - Disposal and disease rates in 340 British dairy herds. AB - Data derived from 340 dairy herds, mainly in southern England, between April 1998 and March 1999, showed that the average total culling rate was 22.1 per cent, with 5.6 per cent for infertility, 3.6 per cent for mastitis, 1.7 per cent for lameness, 2.0 per cent for poor milk yield, 3.7 per cent for age and 5.5 per cent for miscellaneous reasons which included death. The average annual rate of assisted calvings was 8.7 per cent, of injury 0.9 per cent, digestive disease 1.3 per cent ketosis 0.4 per cent, hypomagnesaemia 0.7 per cent, hypocalcaemia 5.3 per cent, mastitis 36.6 per cent, and lameness 23.7 per cent. There was a significant association (P<0.001) between higher rates of mastitis in cows housed in straw yards as opposed to cubicles and also between higher rates of lameness in cows housed in cubicles as opposed to yards (P<0.015). However, there were farms with low rates of mastitis in cows kept in straw yards and low rates of lameness in cows kept in cubicles. Larger herds tended to have more problems with lameness and higher bulk milk somatic cell counts (BMSCC). There was a positive association between BMSCC and mastitis rate. PMID- 10803982 TI - Immunohistochemical detection of p53 and p21 proteins in canine testicular tumours. AB - The objective of this study was to detect by immunohistochemical means nuclear accumulations of p53 and p21 proteins in testicular tumours of dogs. Intense p53 protein nuclear labelling was shown by each of seven seminomas and one intermediate collision tumour. Moderate or intense immunoreactivity was shown by three Sertoli cell tumours. All the tumours also showed p21 nuclear reactivity in parallel with the p53 reactivity. In contrast, four Leydig cell tumours showed no detectable immunoreactivity. The results suggested that high levels of p53 accumulation were associated with the expression of wild-type p53, which was able to activate the transcription of the p21 gene. In addition, weak p53- and p21 nuclear reactivities were detectable in primary spermatocytes within normal seminiferous tubules. PMID- 10803981 TI - Application of an indirect ELISA to milk samples to identify cows with Mycoplasma bovis mastitis. AB - An indirect ELISA was used to detect antibodies to Mycoplasma bovis in milk samples collected from a herd with M bovis mastitis. Antibodies were detected in samples from nine cows which had developed clinical M bovis mastitis. Milk from only three consistently antigen-negative cows tested positive for M bovis antibodies. These results indicate the potential value of the indirect ELISA for the detection of cows which have recently developed M bovis mastitis during the early stages of an outbreak. PMID- 10803984 TI - Symmetrical lupoid onychodystrophy in a crossbred pointer dog: long-term observations. PMID- 10803983 TI - Resection of the common digital extensor tendon in a gelding. AB - A four-year-old gelding was lame owing to a chronic septic common digital extensor tendon and sheath. The horse had been treated by open surgical lavage but the sepsis had recurred after three months. Physical, ultrasonographic, cytological and histological examinations confirmed chronic septic tenosynovitis and tendonitis. The entire intrathecal component of the common digital extensor tendon was resected under general anaesthesia and the synovial lining of the sheath was ablated. Postoperatively the horse regained good limb function and became sound. PMID- 10803985 TI - Persistent efficacy of doramectin pour-on against Haematobia irritans in cattle. PMID- 10803986 TI - Safety and efficacy of a recombinant FeLV vaccine combined with a live feline rhinotracheitis, calicivirus and panleukopenia vaccine. PMID- 10803987 TI - Costs of procedures. PMID- 10803988 TI - Costs of procedures. PMID- 10803989 TI - Hands-on CPD for rural practice? PMID- 10803990 TI - Copper toxicosis in Bedlington terriers. PMID- 10803991 TI - Microbial flora of the abomasal glands of weaned calves. PMID- 10803992 TI - Videoendoscopic thyroidectomy: experimental development of a new technique. AB - Applications of minimally invasive surgery have evolved from laparoscopic cholecystectomy and spread beyond conventional body cavities to new applications and disease processes. The purpose of the present study was to evaluate the feasibility of performing videoendoscopic thyroidectomy in a canine model. Adult mongrel dogs served as the experimental model for this study. The pretracheal space was entered via a 2.5-cm midline neck incision superior to the sternal notch and then expanded with a balloon spacemaker. A mechanical lift device was used to maintain exposure, and a 12-mm midline cervical port and two lateral 5-mm cervical ports were placed. Dissection was carried out with pediatric endoscopic instruments, and hemostasis was obtained with endoscopic clips and an ultrasonic coagulator. Histologic analysis of the excised tissue was performed in all cases. Mean operative time for thyroid lobectomy was 69 minutes (range 58 to 99). The laryngeal nerves and external parathyroid glands were identified endoscopically in all cases. Blood loss was minimal. No complications were observed intraoperatively or during 1 week of postoperative follow-up. Serum calcium levels were stable. No wound hematomas, laryngeal nerve injuries, or other complications occurred. On average 16 mL (range 2 to 65) of seroma fluid was present in the pretracheal space at autopsy. In conclusion, a videoendoscopic approach to thyroid lobectomy is feasible in an animal model and provides adequate visualization of the important anatomy relevant to this procedure. These techniques may be applicable to thyroidectomy in humans and may impact on minimally invasive surgery in other nontraditional locations. PMID- 10803993 TI - Photodynamic therapy for treatment of malignant dysphagia. AB - Photodynamic therapy (PDT) was recently approved by the Food and Drug Administration for palliating obstructing esophageal cancer. This report reviews our initial experience using PDT to treat malignant dysphagia. Patients with inoperable, obstructing esophageal cancer were considered for PDT. Photofrin was injected 48 hours before endoscopic laser activation. Dysphagia score was assessed. Thirty patients underwent 53 PDT courses. Improvement in dysphagia occurred in 83%. Mean dysphagia score decreased from 2.8 to 1.8 (p < 0.05). Complications included esophageal stricture (9.4%), candida esophagitis (5.7%), symptomatic pleural effusion (5.7%), contained esophageal perforation (1.9%), aspiration pneumonia (1.9%), and sunburn (13.2%). Seventeen patients (57%) required more than one PDT treatment, and in 10 an expandable metal stent was used as an adjunct. The 30-day mortality rate was 7%. PDT is effective in palliating patients with malignant dysphagia. The ideal patient for PDT has an obstructing, primarily endoluminal esophageal tumor with minimal extrinsic compression. PMID- 10803994 TI - Effect of open and laparoscopic surgery on cellular immunity in a swine model. AB - Immune suppression associated with trauma has been demonstrated to be proportional to the magnitude of injury. Laparoscopic surgery has been shown to produce a diminished stress response compared to open surgery. Postoperative immune function, specifically cellular immunity, may be better preserved after laparoscopic surgery compared to open surgery. The aim of this study was to examine the effect of open versus laparoscopic surgery on cellular immunity in a swine model. Twenty domestic female pigs were randomly selected for laparoscopic cholecystectomy (LC) or open cholecystectomy (OC). Cellular immune functions were evaluated with delayed-type hypersensitivity (DTH) skin test and serial phytohemoagglutinin (PHA)-induced T-cell proliferation of the peripheral blood. There was a significant reduction in PHA-induced T-cell proliferation in both LC and OC groups on days 1 and 3 compared to preoperative values (p < 0.05). The reduction of mitogen-induced T-cell proliferation after LC was significantly less than after OC on day 1 (p = 0.03). The mean DTH reaction was 29.7 +/- 3.7 mm2 in the LC group compared to 13.9 +/- 1.2 mm2 in the OC group (p < 0.001). There was no difference in postoperative white blood count values between the two groups. Suppression of cellular immunity occurred after both LC and OC. The magnitude and duration of impaired cellular immunity after laparoscopic surgery was less than after open surgery as measured by T-cell proliferation and DTH response. PMID- 10803995 TI - Audit of laparoscopic cholecystectomy in a single center. AB - A continuous audit is required to ensure laparoscopic cholecystectomy (LC) is performed safely in the surgical community in general. A retrospective review of all LC done in a single center was performed. A total of 1,244 LC were attempted. The conversion rate was 12.4%, the complication rate 3.5%, and the bile duct injury rate 0.4%. Forty percent of bile duct injury occurred after conversion. A decreasing trend of complication rate was seen in the early part of the series, then the rate steadied at about 2.5-3%. A higher threshold of conversion may not increase the bile duct injury rate. However, good laparoscopic technique and adequate experience are prerequisites to safe LC. PMID- 10803996 TI - Urinary catheter in laparoscopic cholecystectomy: is it necessary? AB - The aim of this prospective study was to evaluate the necessity or urinary catheterization in elective laparoscopic cholecystectomy. From April 1996 to April 1998, 261 patients undergoing elective laparoscopic cholecystectomy at a county hospital were randomized to either receive or not receive preoperative urinary bladder catheterization. Data analyzed included age and gender of patients, length of surgery, and intraoperative and perioperative complications such as visceral injury, urinary tract infection, and urinary retention. Our results showed, although not statistically significant, more urinary tract complications in the "with Foley" group than in the "without Foley" group (four vs one, respectively). There was no significant difference between the two groups with respect to length of operation and perioperative complications. There was no visceral injury or operative mortality in this study. We conclude that urinary catheterization can be omitted safely in elective laparoscopic cholecystectomy. PMID- 10803997 TI - Laparoscopic versus open appendectomy: prospective randomized trial. AB - Although classic open surgery is simple, expeditious, and effective, it has some drawbacks, including wound sepsis, delayed recovery, operative difficulties, and possibility of unnecessary appendectomies for false appendicitis. The aim of this study was to assess the applicability and safety of laparoscopic appendectomy (LA) in a prospectively randomized trial. Seventy nonselective patients with suspected appendicitis were randomized to laparoscopic (n = 35, 17 male) or open appendectomy (n = 35, 15 male) and operated on an emergency basis. Operative findings, operating time, postoperative complications, and length of hospital stay were compared. We found that LA is associated with a shorter hospital stay, fewer postoperative complications, and better diagnostic accuracy, and it is recommended as the procedure of choice for the diagnosis and management of acute appendicitis. PMID- 10803998 TI - Preperitoneal hernioplasty performed with needlescopic instruments (microlaparoscopy). AB - We treated a series of 30 patients who had a Nyhus type II, III, or IV inguinal hernia without associated pathology by laparoscopic surgery using 2-mm surgical instruments and a modified preperitoneal Dulucq technique. A 30 degrees optic was used, and the repair was reinforced by an accordion-folded, polypropylene mesh introduced through a 10-mm optic needleport. There were no postoperative complications, and all patients were discharged 24 hours after operation. At 6 months there has been no recurrence of hernia. It was evident that the 2-mm procedure produced less surgical trauma. PMID- 10803999 TI - Intrasphincteric botulinum toxin type A for the diagnosis of sphincter of Oddi dysfunction: a case report. AB - Biliary-type pain from sphincter of Oddi dysfunction is not uncommon after cholecystectomy. An increased basal pressure of sphincter of Oddi manometry establishes the diagnosis and treatment is usually by endoscopic sphincterotomy. Both procedures carry a significant complication rate. A few patients with elevated sphincter pressure do not respond to therapy; the source of their pain may be elsewhere. This case report describes the use of intrasphincteric botulinum toxin injection for the diagnosis of sphincter of Oddi dysfunction in a patient after repeated attempts at manometry had failed. This may provide a safe and easy method of determining whether sphincter of Oddi dysfunction may be the cause of biliary pain in post/ cholecystectomy patients and help select patients who would benefit from subsequent sphincter ablation, without the risks of sphincter of Oddi manometry. Prospective studies are first needed. PMID- 10804000 TI - Laparoscopically assisted splenectomy following preoperative splenic artery embolization using contour emboli for myelofibrosis with massive splenomegaly. AB - Laparoscopically assisted splenectomy with an 8- to 10-cm left upper paramedian laparotomy was performed following preoperative splenic artery embolization using painless contour emboli (super absorbent polymer microsphere) with early successful results in two men (46 and 37 years old) with myelofibrosis accompanied by massive splenomegaly. Dissection around the lower part of the spleen and the hilum initially was performed intracorporeally with the usual laparoscopic view under 12 mm Hg pneumoperitoneum. The alternating changes of viewpoints between the direct view through an 8- to 10-cm incision and the usual laparoscopic view with or without application of a retraction method were effective for safe hilar devascularization. Preoperative splenic artery embolization at the distal site was effective for safe dissection around the enlarged spleen. The patients did not complain of pain before operation. Preoperative painless embolization and laparoscopically assisted splenectomy with small laparotomy promotes the feasibility and safety of minimally invasive splenectomy for myelofibrosis with massive splenomegaly. PMID- 10804001 TI - Combined laparoscopic splenectomy and cholecystectomy for the treatment of hereditary spherocytosis: is it safe and effective? AB - Hereditary spherocytosis is the most common red blood cell membrane disorder and often is associated with hemolytic crisis and premature cholelithiasis. Splenectomy is the only effective therapy for this disorder and often it is performed in combination with cholecystectomy. Conventional surgery requires a wide upper abdominal incision for correct exposure of the gallbladder and spleen. Laparoscopic cholecystectomy and splenectomy have been performed safely worldwide. We report our experience with seven patients (one male and six female, average age 12 years) who underwent combined laparoscopic splenectomy and cholecystectomy for hereditary spherocytosis. The patient was placed in supine position and the procedure performed with a five-trocar technique. Cholecystectomy was performed first, then splenectomy was achieved and the spleen removed by morcellation into a retrieval bag (five cases) or via a 4- to 5-cm left subcostal incision (two cases). No patient required conversion to open technique or blood transfusion. The mean blood loss was 162 mL, mean operative time 207 minutes, mean spleen size 14.5 cm, and median postoperative hospital stay 4 days. No perioperative mortality or major complications occurred in our series. After a median follow-up of 18 months all patients showed sharp hematologic improvement. Despite the small number of cases, we consider the combined laparoscopic approach safe and effective for the treatment of hereditary spherocytosis. PMID- 10804002 TI - Primary intrahepatic lithiasis: report of a case treated by laparoscopic bilioenteric anastomosis. AB - Recent advances in videolaparoscopic surgery have made this method the treatment of choice for many biliary diseases. However, it has not been used in certain cases, such as primary intrahepatic lithiasis. The authors report a case of a 62 year-old woman with a history of several episodes of cholangitis. Investigation revealed dilated intra- and extrahepatic bile ducts with intrahepatic stones. The patient underwent laparoscopy, and intraoperative cholangiography disclosed an enlarged common duct with absence of stones and the presence of multiple calculi in the intrahepatic biliary tree. A choledochotomy followed by choledochoscopy was performed, which revealed several intrahepatic pigmented stones that were completely retrieved, followed by a laterolateral choledochoduodenostomy to decompress the biliary tree and to allow the migration of residual or recurrent stones. The patient had an uneventful recovery and was discharged on the fourth postoperative day. After 15 months of follow-up the patient is asymptomatic with normal results of liver function tests. Late postoperative upper digestive endoscopy showed a patent choledochoduodenostomy. PMID- 10804003 TI - Anomalous insertion of the right hepatic duct into the cystic duct: report of a case diagnosed before laparoscopic cholecystectomy. AB - Although an aberrant hepatic duct entering the cystic duct is not especially rare, the main right hepatic duct entering the cystic duct is extremely rare, with only six cases reported thus far. All of the reported patients underwent open cholecystectomy, during which one patient received a bile duct injury. The anomaly was unsuspected preoperatively in all of these cases. We report an additional patient with this anomaly, the first such case diagnosed before laparoscopic cholecystectomy using direct cholangiography. Cholangiography may be mandatory whenever biliary anomalies are suspected during laparoscopic cholecystectomy. As the right hepatic duct entering the cystic duct can lead to ductal injury, this anomaly should be kept in mind when performing laparoscopic cholecystectomy. PMID- 10804004 TI - Treatment of cholecystoduodenal fistula by laparoscopy. AB - We describe a patient with cholecystoduodenal fistula treated by a laparoscopic approach. Use of a flexible videoscope, flexible retractor, and endoscopic transecting stapler allows for laparoscopic treatment of cholecystoenteric fistulae. PMID- 10804005 TI - Laparoscopic treatment of cholecystocolonic fistula: report of a case preoperatively diagnosed by barium enema. AB - The authors present a case of cholecystocolonic fistula with no specific symptoms, such as severe diarrhea or pneumobilia, preoperatively diagnosed and treated by the laparoscopic approach. A preoperative barium enema demonstrated a cholecystocolonic fistula. The fistula was divided by the laparoscopic stapling technique. Important features in the management of this case are (1) preoperative diagnosis of the fistula by barium enema carried out for screening colorectal cancer, (2) dissection of the gallbladder from its bed before division of the fistula, and (3) use of the laparoscopic stapling technique to divide the fistula while preventing fecal soilage. PMID- 10804006 TI - Cologastric fistula and colonic perforation as a complication of percutaneous endoscopic gastrostomy. AB - Cologastric fistula has rarely been reported as a complication of percutaneous endoscopic gastrostomy (PEG). We encountered a patient in whom this problem went unrecognized for 2 years. After the initial PEG tube was changed, the second PEG tube was advanced into the colon, causing severe diarrhea. When a third PEG tube was inserted, acute peritonitis occurred because of colonic perforation. We discuss the mechanism of this complication and technical points related to its prevention. PMID- 10804007 TI - Laparoscopic removal of fish bone. AB - A patient swallowed a fish bone that perforated the stomach wall and embedded in the head of the pancreas. This was confirmed by computed tomographic imaging. We describe a case of successful fish bone removal using a laparoscopic technique, thus avoiding a laparotomy. The patient recovered well and was discharged from the hospital the next day. This is the first description of such a case. PMID- 10804008 TI - Small bowel perforation caused by a sharp bone: laparoscopic diagnosis and treatment. AB - Small bowel perforation caused by sharp or pointed foreign bodies rarely is diagnosed preoperatively because clinical symptoms usually are nonspecific and can mimic other surgical conditions such as appendicitis and cecal diverticulitis. We report the case of a 57-year-old man who underwent diagnostic laparoscopy for abdominal pain of unknown etiology. Swelling and erythema were noted in a segment of small bowel in the right lower abdomen. A tiny sharp pointed object was found penetrating the inflamed portion of the bowel. This foreign body was removed and the perforation immediately repaired under laparoscopic guidance. The postoperative course was uneventful. We believe that in cases of lower abdominal pain with peritoneal irritation, laparoscopy can provide not only a wide field of examination to localize intraabdominal pathology, but in some instances it can aid in case management. PMID- 10804009 TI - Laparoscopic omentectomy for peritoneal dialysis catheter flow obstruction: a case report and review of the literature. AB - Peritoneal dialysis catheter outflow dysfunction frequently is caused by omentum or epiploic appendices blocking the side holes of the catheter tubing. Laparoscopy has been utilized increasingly to remedy mechanical blockage of the peritoneal device. This obviates the need for catheter replacement, permits early return to dialysis, and lowers the patient dropout rate from peritoneal dialysis as a modality for renal replacement therapy. We report a case of laparoscopic subtotal omentectomy to successfully resolve recurrent catheter obstruction that failed previous omentolysis and omentopexy. Using four port sites, the omental resection was accomplished using the harmonic scalpel. The procedure was performed on an ambulatory basis, dialysis was resumed after 1 week, and there were no infectious complications or port site leaks. We define terminology to standardize reporting of omentectomy. Our technique of laparoscopic omentectomy is described and compared to those of previously published reports. The role of laparoscopic omentectomy relative to other commonly used laparoscopic approaches to the obstructed peritoneal dialysis catheter is reviewed. PMID- 10804010 TI - Laparoscopic repair of pleural laceration produced during truncal vagotomy: case report. AB - A partial pneumothorax developed in a patient undergoing laparoscopic truncal vagotomy when a small pleural laceration was accidentally produced. Changes in oxygen saturation and PETCO2 were immediately detected by the anesthesiologist and measures were taken to maintain the patient's ventilatory stability. The pleural laceration was repaired laparoscopically, and the pneumothorax was corrected by ventilatory manipulation, avoiding the placement of a chest tube. The procedure was completed uneventfully. Literature about the causes of pneumothorax during laparoscopic procedures as well as preventive and therapy viewed. PMID- 10804011 TI - Liver retractor for laparoscopic fundoplication. PMID- 10804012 TI - Increased expression of bone sialoprotein in bone metastases compared with visceral metastases in human breast and prostate cancers. AB - The recent demonstration that bone sialoprotein (BSP) is expressed in osteotropic cancers suggests that this bone matrix protein might be implicated in the preferential seed and growth of metastatic cells in bone. High expression of BSP in breast and prostate primary carcinomas is associated with progression and bone metastases development. The exact mechanisms by which BSP may favor bone metastases formation are not clearly established yet. Although BSP expression has been detected in breast, prostate, lung, thyroid, and neuroblastoma primary tumors, no information regarding its expression in metastases is available to date. In this study, we have examined BSP expression in 15 bone and 39 visceral metastatic lesions harvested from 8 breast cancer patients and 7 prostate cancer patients who died of disseminated disease. We were able to retrieve the primary lesions from 5 of the 8 breast cancer patients as well as from all 7 prostate cancer patients. All the primary breast tumor patients and 5 of the 7 primary prostate cancer patients expressed a detectable level of BSP. Bone metastases from all 8 breast cancer patients and from 5 out of 7 prostate cancer patients exhibited detectable levels of the protein. Metastatic cells in close contact with bone trabeculae usually were highly positive for BSP. BSP also was detected in secondary lesions developed at visceral sites including liver, thyroid, lung, and adrenal glands. However, BSP expression was significantly lower in visceral metastases than in skeletal ones (Mann-Whitney test, p < 0.05). Our data represent the first demonstration of an increased expression of BSP in bone metastases compared with nonskeletal metastases in human breast and prostate cancers and add weight to the body of evidence attributing a significant role to this protein in the genesis of bone metastases. PMID- 10804013 TI - Pathogenesis of bone metastases: role of tumor-related proteins. PMID- 10804014 TI - Increased marrow-derived osteoprogenitor cells and endosteal bone formation in mice lacking thrombospondin 2. AB - The phenotype of thrombospondin 2 (TSP2)-null mice includes abnormalities in collagen fibrils and increases in ligamentous laxity, vascular density, and bleeding time. In this study, analyses by computerized tomography (CT) revealed that cortical density was increased in long bones of TSP2-null mice. Histomorphometric analysis showed that the mid-diaphyseal endosteal bone formation rate (BFR) of TSP2-null mice was increased in comparison with that of wild-type (WT) animals. Although microgeometric analysis showed that periosteal and endosteal radii were reduced, the mechanical properties of femurs from TSP2 null mice were not significantly different from those of controls, presumably because of the concomitant increase in endosteal bone mass. Bone loss in ovariectomized mice was equivalent for WT and mutant mice, a finding that indicates that TSP2-null animals are capable of normal bone resorption. To further explore the cellular basis for the increased endosteal BFR in TSP2-null mice, marrow stromal cells (MSCs) were isolated and examined in vitro. These cells were found to be present in increased numbers in a colony forming unit (CFU) assay and showed an increased rate of proliferation in vitro. We conclude that TSP2 regulates the proliferation of osteoblast progenitors, directly or indirectly, and that in its absence endosteal bone formation is increased. PMID- 10804015 TI - In vivo demonstration that human parathyroid hormone 1-38 inhibits the expression of osteoprotegerin in bone with the kinetics of an immediate early gene. AB - Osteoprotegerin (OPG) is a potent inhibitor of osteoclast formation and function. To elucidate how OPG is regulated in bone, we examined (1) the expression and localization of OPG protein in bone tissue, (2) the effect of human parathyroid hormone 1-38 (hPTH 1-38) on OPG messenger RNA (mRNA) levels in rat femur metaphyseal and diaphyseal bone, and (3) the effect of hPTH(1-38) on expression of OPG mRNA in cultured osteoblast-like cells derived from the metaphysis and diaphysis, and in ROS 17/2.8 osteosarcoma cells. Because PTH has been shown to stimulate osteoblast activity via the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal transduction pathway we also investigated whether PTH action on OPG in vivo is dependent on activation of cAMP/PKA pathway. Immunohistochemistry was used to evaluate OPG protein expression and Northern blot hybridization was used to analyze OPG mRNA expression both in vivo and in vitro. Immunohistochemistry of OPG protein expression in the rat distal femur metaphysis revealed that it was localized predominantly in preosteoblasts, osteoblasts, lining cells, and the osteoid layer, with occasional immunoreactivity in osteocytes and cells of the bone marrow. Subcutaneous (sc) administration of a single injection of hPTH(1-38) at 80 microg/kg induced a rapid and transient decrease in OPG mRNA expression in both metaphyseal and diaphyseal bone. The decrease in OPG message was evident by 1 h and mRNA levels returned to baseline after 3 h. PTH analog PTH(1-31), which stimulates intracellular cAMP accumulation, inhibited OPG expression, whereas PTH analogs (3 34 and 7-34) that do not stimulate cAMP production had no effect on expression. In contrast to PTH, prostaglandin E2 (PGE2) had no effect on OPG mRNA expression in vivo in the metaphyseal bone cells, under conditions in which PGE2 does promote expression of the c-fos gene. The in vivo effects of hPTH(1-38) on OPG mRNA were confirmed in isolated primary osteoblast cultures derived from either metaphyseal or diaphyseal bone as well as in ROS 17/2.8 osteosarcoma cells. We propose that the rapid and transient decrease in OPG expression may initiate a cascade of events resulting in the differentiation of osteoclast progenitor. Such a spatially and temporally programmed effect of PTH might contribute to bone turnover. PMID- 10804016 TI - Diet- or warfarin-induced vitamin K insufficiency elevates circulating undercarboxylated osteocalcin without altering skeletal status in growing female rats. AB - To further characterize the skeletal role of vitamin K (K), markers of bone turnover, density, and strength were evaluated in rats with diet- or warfarin (W) induced K insufficiency. One hundred two, 7-week-old, female rats were randomly assigned to low K (phylloquinone [K1], 20 microg/kg diet), control K (K1, 1300 microg/kg diet), low-dose W (W, 1.5 mg/kg control diet), or high-dose W plus K (W/K1, 10/100 mg/kg diet). Femur bone mineral content (BMC) and bone mineral density (BMD), plasma prothrombin time (PT) and prothrombin concentration (PC), and serum total alkaline phosphatase (ALP) and skeletal alkaline phosphatase (sALP) were measured at baseline and days 20, 40, 60, and 80. Serum total osteocalcin (OC) and undercarboxylated osteocalcin (ucOC) and femur length (FL) were measured at baseline and day 80. Left femur OC was measured and biomechanical testing of the right femur and third lumbar vertebral body was performed at day 80. Low dietary K elevated circulating ucOC (17% higher than control; p < 0.0001) at day 80. Furthermore, in both W groups, essentially all circulating OC was undercarboxylated and femur OC was lower than control (p < 0.0001). However, there was no change in femur percent ucOC, suggesting deposition of less newly synthesized OC. No between group differences were observed in PT, ALP, sALP, FL, BMC, BMD, or bone strength. In conclusion, skeletal K insufficiency can be induced by W or diet manipulation. This does not hinder peak bone mass attainment in female rats; however, W causes less newly synthesized OC to be deposited in bone. PMID- 10804018 TI - Protein kinase C involvement in interleukin-6 production by parathyroid hormone and tumor necrosis factor-alpha in UMR-106 osteoblastic cells. AB - The cytokine interleukin-6 (IL-6) is increased in bone and bone cells by several resorptive stimuli, including parathyroid hormone (PTH), IL-1beta, and tumor necrosis factor-alpha (TNF-alpha). The current studies were designed to determine the contribution of the protein kinase C (PKC) signaling pathway to the effects of these three agents to increase IL-6 in UMR-106 rat osteoblastic cells. Cells were pretreated with vehicle (dimethylsulfoxide [DMSO]) or the phorbol ester, phorbol 12,13-dibutyrate (PDB; 300 nM) for 48 h to down-regulate phorbol sensitive PKC isozymes. Either PTH (0.1-10 nM), IL-1beta (0.1-10 nM), or TNF alpha (5 nM and 10 nM) was then added for 24 h in the continued presence of vehicle or PDB. PKC isozymes were visualized by Western immunoblotting and IL-6 was determined by bioassay. PDB pretreatment caused a partial down-regulation of the conventional alpha-PKC and betaI-PKC isozymes and complete down-regulation of the novel delta-isoenzyme and epsilon-isozymes but it had no effect on the atypical zeta-PKC isozyme. PDB pretreatment reduced IL-6 responses to 5 nM and 10 nM PTH by 61% and 33%, respectively, reduced IL-6 responses to 5nM and 10 nM TNF a by 54% and 42%, respectively, and failed to inhibit the IL-6 responses to 0.1 10 nM IL-1beta. The PDB pretreatment protocol significantly enhanced PTH stimulated cyclic adenosine monophosphate (cAMP) production. The PKC inhibitor calphostin C also decreased IL-6 responses to PTH. Thus, in this osteoblast cell line, the PKC pathway is an important component of the signaling pathway for the IL-6 production stimulated by PTH and TNF-alpha but not that from IL-1beta. PMID- 10804019 TI - Thiazide diuretics affect osteocalcin production in human osteoblasts at the transcription level without affecting vitamin D3 receptors. AB - Besides their natriuretic and calciuretic effect, thiazide diuretics have been shown to decrease bone loss rate and improve bone mineral density. Clinical evidence suggests a specific role of thiazides on osteoblasts, because it reduces serum osteocalcin (OC), an osteoblast-specific protein, yet the mechanisms implicated are unknown. We therefore investigated the role of hydrochlorothiazide (HCTZ) on OC production by the human osteoblast-like cell line MG-63. HCTZ dose dependently (1-100 microM) inhibited 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] induced OC release by these cells (maximal effect, -40-50% and p < 0.005 by analysis of variance [ANOVA]) as measured by ELISA. This effect of HCTZ on OC release was caused by a direct effect on OC gene expression because Northern blot analysis revealed that OC messenger RNA (mRNA) levels were reduced in the presence of increasing doses of the diuretic (-47.2+/-4.0%; p < 0.0001 by paired ANOVA with 100 microM 13.6+/-0.49 pmol/mg protein/15 minutes; p < 0.05) in MG-63 cells. Reducing extracellular Ca2+ concentration with 0.5 mM EDTA or 0.5 mM ethylene glycol-bis(beta-amino ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) only partly prevented the inhibitory effect of the diuretic on OC secretion (maximal effect, -22.5+/-6.9%), suggesting that thiazide-dependent Ca2+ influx is not sufficient to elicit the inhibition of OC secretion. Because OC production is strictly dependent on the presence of 1,25(OH)2D3 in human osteoblasts, we next evaluated the possible role of HCTZ on vitamin D3 receptors (VDR) at the mRNA and protein levels. Both Northern and Western blot analyses showed no effect of HCTZ (1-100 microM) on VDR levels. The presence of EGTA in the culture media reduced slightly the VDR mRNA levels under basal condition but this was not modified in the presence of increasing levels of HCTZ. The OC gene promoter also is under the control of transcription factors such as Yin Yang 1 (YY1) and cFOS. Western blot analysis revealed no changes in YY1 levels in response to HCTZ either in the presence or in the absence of 0.5 mM EGTA in the culture media. In contrast, HCTZ induced a dose-dependent increase in cFOS levels (p < 0.002 by ANOVA), a situation prevented by incubation with EGTA. These studies indicate that HCTZ inhibits OC mRNA expression independently of an effect on VDR, YY1, or extracellular Ca2+ levels but involves changes in cFOS levels. As OC retards bone formation/mineralization, the inhibition of OC production by HCTZ could explain its preventive role in bone loss rate. PMID- 10804017 TI - Parathyroid hormone regulates transforming growth factor beta1 and beta2 synthesis in osteoblasts via divergent signaling pathways. AB - Parathyroid hormone 1-34 [PTH(1-34)] was shown to increase transforming growth factor beta1 (TGF-beta1) and TGF-beta2 concentrations in supernatants of cultured human osteoblasts and to increase TGF-beta1 and TGF-beta2 messenger RNA (mRNA) concentrations and gene transcription in these cells. Because PTH(1-34) activates both protein kinase C (PKC) and protein kinase A (PKA) pathways in osteoblasts, we investigated the role of each kinase pathway in activation of TGF-beta isoforms. PTH(29-32), which activates the PKC pathway in rat osteoblasts, increased TGF-beta1 but not TGF-beta2 concentrations in supernatants of osteoblasts. Phorbol myristate acetate (PMA), a PKC agonist, increased TGF-beta1 but not TGF-beta2 concentrations. Specific PKC antagonists safingol and Go6976 attenuated PTH(1-34)-mediated increases in TGF-beta1 but not TGF-beta2 synthesis. PTH(1-31), which increases PKA activity in several cell culture systems, increased TGF-beta2 but not TGF-beta1 concentrations in human osteoblast supernatants. Forskolin, a PKA agonist, increased TGF-beta2 but not TGF-beta1 concentrations in supernatants of human osteoblasts. The PKA antagonist H-89 blunted PTH(1-34)-mediated increases in TGF-beta2 but not TGF-beta1 synthesis. Our results are consistent with the concept that PTH increases TGF-beta1 expression and secretion by pathways that involve the PKC pathway, whereas it increases TGF-beta2 expression and secretion via the PKA pathway. PMID- 10804020 TI - Stimulatory effect of insulin-like growth factor binding protein-5 on mouse osteoclast formation and osteoclastic bone-resorbing activity. AB - Insulin-like growth factor binding protein-5 (IGFBP-5) stimulates osteoblast proliferation directly or indirectly through IGF-I action, but its effects on osteoclast formation and osteoclastic activity are unknown. We tested the effects of IGFBP-5 on osteoclastic activity and osteoclast formation. IGFBP-5 significantly stimulated pit formation by pre-existent osteoclasts in mouse bone cell cultures and its stimulatory effect was completely blocked by IGF-I antibody (Ab). However, IGFBP-5 did not affect the bone-resorbing activity of isolated rabbit osteoclasts. When IGFBP-5 was added to unfractionated bone cells after degeneration of pre-existent osteoclasts, IGFBP-5 (77 pM-7.7 nM) dose-dependently stimulated osteoclast-like cell formation, irrespective of the presence of IGF-I Ab. Moreover, osteoclast-like cells newly formed by IGFBP-5 from unfractionated bone cells possessed the ability to form pits on dentine slices. We next examined the direct effect of IGFBP-5 on osteoclast precursors in the absence of stromal cells, using hemopoietic blast cells derived from spleen cells. IGFBP-5 dose dependently stimulated osteoclast-like cell formation from osteoclast precursors, irrespective of the presence of IGF-I Ab. Growth hormone (GH) as well as IGF-I significantly stimulated bone resorption by pre-existent osteoclasts in mouse bone cell cultures and these stimulatory effects were completely blocked by IGF-I Ab. GH as well as IGF-I stimulated osteoclast-like cell formation from unfractionated bone cells and this stimulatory effect of GH was significantly but partially blocked by IGF-I Ab. The direct stimulatory effect of GH on osteoclast like cell formation from hemopoietic blast cells was not affected by IGF-I Ab. The present data indicate that IGFBP-5 stimulates bone resorption both by stimulation of osteoclast formation in an IGF-I-independent fashion and by IGF-I dependent activation of mature osteoclasts, possibly via osteoblasts, in vitro. PMID- 10804021 TI - The inhibitory effect of interleukin-10 on mouse osteoclast formation involves novel tyrosine-phosphorylated proteins. AB - Interleukin-10 (IL-10) inhibits osteoclast (OC) formation in rat and mouse systems. However, little is known concerning the mechanism of this inhibitory effect. Using a coculture system of mouse bone marrow cells and primary osteoblastic cells (POB), we evaluated the potential target cells for IL-10 and components of the IL-10 activating pathway. In the coculture system, IL-10 treatment abolished OC differentiation in a dose-dependent manner. This inhibitory effect occurred regardless of the stage of cellular proliferation and differentiation, suggesting that IL-10 may act on a variety of genes participating in OC formation. IL-10 specifically abrogated the production of IL 6 by enriched bone marrow-derived mononuclear cells (BMM) but not by osteoblastic cells. IL-10 treatment also stimulated the binding of a protein in the BMM to an IL-10 response element, whereas no such activation was induced in osteoblastic cells. In contrast, interferon gamma (IFN-gamma), another inhibitory factor, stimulated tyrosine-phosphorylated proteins to bind to an IL-10 response element in both monocytes and osteoblastic cells. These data suggest that the BMM are the direct target of IL-10 action. Importantly, oligonucleotide-specific precipitation confirmed that IL-10 treatment strongly augmented 88, 85, and 70 kDa tyrosine-phosphorylated proteins in BMM. Taken together, these data show that IL-10 inhibits mouse OC formation by acting directly on hemopoietic OC precursor, through a novel signal transduction and activation pathway. PMID- 10804022 TI - Bone-resorbing osteoclasts contain gap-junctional connexin-43. AB - Intercellular gap junctions have been previously described at contact sites between surface osteoblasts, between osteoblasts and underlying osteocytes, and between osteocyte cell processes in the canaliculi. The subunits of gap junction channels are assembled from a family of proteins called connexins. In the present work, we show that rat osteoclasts cultured on bovine bone slices show connexin 43 (Cx43) staining localizing in the plasma membrane of the cells in cell-cell contacts and over the basolateral membrane of osteoclasts. The effect of heptanol, a known gap-junctional inhibitor, was studied using the well characterized pit formation assay. Heptanol decreased the number and activity of osteoclasts. The proportion of mononuclear tartrate-resistant acid phosphatase (TRAP)-positive cells out of all TRAP-positive cells increased on heptanol treatment, suggesting a defect in the fusion of mononuclear osteoclast precursors to multinucleated mature osteoclasts. Furthermore, the total resorbed area and the number of resorption pits also decreased in the heptanol-treated cultures. These results suggest that gap-junctional Cx43 plays a functional role in osteoclasts and that the blocking of gap junctions decreases both the number and the activity of osteoclasts. This can indicate both a direct communication between multinucleated osteoclasts and mononuclear cells through gap junctions or an indirect effect through gap junctions between osteoblasts. PMID- 10804023 TI - Effect of a low calcium dialysate on parathyroid hormone secretion in diabetic patients on maintenance hemodialysis. AB - Diabetic patients on maintenance dialysis often are characterized by a relative parathyroid hormone (PTH) deficiency and a form of renal osteodystrophy with low bone turnover known as adynamic bone. The goal of the present study was to determine whether a reduction in the dialysate calcium concentration would increase the predialysis (basal) PTH and maximal PTH level. Thirty-three diabetic maintenance hemodialysis patients with basal PTH values less than 300 pg/ml were randomized to be dialyzed with either a regular (3.0 mEq/liter or 3.5 mEq/liter, group I) or low (2.25 mEq/liter or 2.5 mEq/liter, group II) calcium dialysate for 1 year. At baseline and after 6 months and 12 months of study, low (1 mEq/liter) and high (4 mEq/liter) calcium dialysis studies were performed to determine parathyroid function. At baseline, basal (I, 126+/-20 vs. II, 108+/-19 pg/ml) and maximal (I, 269 pg/ml+/-40 pg/ml vs. II, 342 pg/ml+/-65 pg/ml) PTH levels were not different. By 6 months, basal (I, 98+/-18 vs. II, 200+/-34 pg/ml, p = 0.02) and maximal (I, 276 pg/ml+/-37 pg/ml vs. II, 529 pg/ml+/-115 pg/ml; p = 0.05) PTH levels were greater in group II. Repeated measures analysis of variance (ANOVA) of the 20 patients who completed the entire 12-month study showed that only in group II patients were basal PTH (p = 0.01), maximal PTH (p = 0.01), and the basal/maximal PTH ratio (p = 0.03) different; by post hoc test, each was greater (p < 0.05) at 6 months and 12 months than at baseline. When study values at 0, 6, and 12 months in all patients were combined, an inverse correlation was present between basal calcium and both the basal/maximal PTH ratio (r = -0.59; p < 0.001) and the basal PTH (r = -0.60; p < 0.001). In conclusion, in diabetic hemodialysis patients with a relative PTH deficiency (1) the use of a low calcium dialysate increases basal and maximal PTH levels, (2) the increased secretory capacity (maximal PTH) during treatment with a low calcium dialysate suggests the possibility of enhanced parathyroid gland growth, and (3) the inverse correlation between basal calcium and both the basal/maximal PTH ratio and the basal PTH suggests that the steady-state PTH level is largely determined by the prevailing serum calcium concentration. PMID- 10804024 TI - Selective drug delivery system to bone: small peptide (Asp)6 conjugation. AB - Targeting a drug on hydroxyapatite (HA) could be a promising way for selective drug delivery to bone, because HA, an inorganic component in hard tissues (bone and teeth), does not exist in soft tissues. Several bone noncollagenous proteins, which bind to HA, have repeating sequences of acidic amino acids in their structures as possible HA-binding sites. Thus, we think that a small peptide of repetitive acidic amino acid could work as a carrier for selective drug delivery to the bone. To test this hypothesis, we conjugated (Asp)6 to fluorescein isothiocyanate (FITC), evaluated its affinity to HA in vitro, and examined its tissue distribution after injection into rats. Although fluorescein itself did not bind to HA, (Asp)6-FITC bound to HA as well as calceine and tetracycline. Twenty-four hours after intravenous injection of (Asp)6-FITC to rats, animals were killed, and ground sections of hard tissues and cryosections of soft tissues were made. Under a confocal laser scanning microscope, clear labeling lines were observed in bones and teeth, whereas no labeling was detected in soft tissues. In the rats administered with fluorescein alone, the fluorescent labeling was detected in neither hard nor soft tissues. Fluorescent analysis of blood, urine, and bones after (Asp)6-FITC administration revealed that biological half-life of FITC in blood was short (60 minutes) and that within 24 h, 95% of the administered FITC was excreted as urine whereas 2% of the FITC accumulated in bones. After subcutaneous administration of (Asp)6-FITC to mice, fluorescent intensity remaining in the femurs was measured periodically. In these mice the biological half-life of FITC in the femur was 14 days. Present results indicate that (Asp)6 is effective as a carrier for selective drug delivery to bone. PMID- 10804025 TI - Bone mass continues to increase at the hip after parathyroid hormone treatment is discontinued in glucocorticoid-induced osteoporosis: results of a randomized controlled clinical trial. AB - Glucocorticoid-induced osteoporosis is the most common secondary cause of osteoporosis. In this 24-month study, we report changes in bone turnover and bone mass after 12 months of daily injections of human parathyroid hormone 1-34 [hPTH(1-34)] and 12 months off treatment in postmenopausal women (mean age, 63 years) with osteoporosis treated with glucocorticoid and hormone replacement therapy. Response to the treatment was assessed with bone mineral density (BMD) measurements of the lumbar spine by quantitative computed tomography (QCT); BMD measurements of the lumbar spine, hip, and forearm by dual-energy X-ray absorptiometry (DXA); and biochemical markers of bone turnover. The mean (+/-SEM) change in BMD of the lumbar spine by QCT and DXA in the PTH group at 24 months was 45.9+/-6.4% and 12.6+/-2.2% (p < 0.001). The change in total hip and femoral neck BMD was not significant at 12 months but increased to 4.7+/-0.9% (p < 0.01) and 5.2+/-1.3% at 24 months, respectively, as compared with a relatively small change of 1.3+/-0.9% and 2.6+/-1.7% in the estrogen-only group. The mean percent differences in BMD of the lumbar spine by QCT and DXA between the groups at 24 months were 43.1% and 11.9%, respectively (p < 0.001). The mean percent differences over the estrogen-only group in hip BMD were 3.4% for total hip (p < 0.01) and 2.6% for femoral neck at 24 months. Biochemical markers of bone turnover increased to more than 150% during the first 6 months of therapy, remained elevated throughout the 12-month treatment period, and returned to baseline values within 6 months of discontinuing the PTH treatment. These results suggest that PTH dramatically increases bone mass in the lumbar spine and hip in postmenopausal women with glucocorticoid-induced osteoporosis who are taking hormone replacement therapy. However, the maximum effect of this anabolic agent on bone mass at the hip after 12 months of treatment requires at least 6-12 months after the PTH treatment is discontinued. PMID- 10804026 TI - Corticosteroids do not alter the threshold for vertebral fracture. AB - Corticosteroid use is one of the most important secondary causes of osteoporosis. Generally, it has been believed that in addition to its effect on bone mineral density (BMD), it also causes an alteration in bone quality that means that fractures occur at a lower BMD than might be expected. To establish if this is the case, we have compared the relationship between BMD and vertebral fracture in patients receiving corticosteroids with that in patients who had never received such therapy. Information was gathered on those patients who had been referred to the participating centers and had both BMD measurements and lateral thoracolumbar radiographs. In all, 452 patients (391 female) were identified; of these 82 (63 female) were receiving corticosteroids. There was no significant difference in BMD between the patients on corticosteroids and those with other suspected causes of osteoporosis. Vertebral fractures were present in 53% of patients on steroids compared with 35% of those who had no such treatment (p = 0.0035). The fractures were more likely to be multiple in patients on corticosteroids (p = 0.0042). However, if the relationship between bone density and fracture is investigated by plotting the cumulative prevalence of fracture against the bone density, measured by T score, the median BMD for fractures actually was marginally lower in patients on steroids, -2.74 (95% confidence interval [CI], -2.77 to -2.70) compared with -2.65 (95% CI, -2.66 to -2.65) in those who had not received steroids. Our results fail to support the notion that the fracture threshold is altered in patients on long-term steroids and suggest that the same diagnostic criteria should be used for osteoporosis in patients whether or not they are taking corticosteroid therapy. PMID- 10804027 TI - Deficient bone formation in idiopathic juvenile osteoporosis: a histomorphometric study of cancellous iliac bone. AB - Idiopathic juvenile osteoporosis (IJO), a rare cause of osteoporosis in children, is characterized by the occurrence of vertebral and metaphyseal fractures. Little is known about the histopathogenesis of IJO. We analyzed by quantitative histomorphometry iliac crest biopsies from 9 IJO patients (age, 10.0-12.3 years; 7 girls) after tetracycline labeling. Results were compared with identically processed samples from 12 age-matched children without metabolic bone disease and 11 patients with osteogenesis imperfecta type I. Compared with healthy controls, cancellous bone volume (BV) was markedly decreased in IJO patients (mean [SD]: 10.0% [3.1%] vs. 24.4% [3.8%]), because of a 34% reduction in trabecular thickness (Tb.Th) and a 37% lower trabecular number (Tb.N; p < 0.0001 each; unpaired t-test). Bone formation rate (BFR) per bone surface was decreased to 38% of the level in controls (p = 0.0006). This was partly caused by decreased recruitment of remodeling units, as shown by a trend toward lower activation frequency (54% of the control value; p = 0.08). Importantly, osteoblast team performance also was impaired, as evidenced by a decreased wall thickness (W.Th; 70% of the control value; p < 0.0001). Reconstruction of the formative sites revealed that osteoblast team performance was abnormally low even before mineralization started at a given site. No evidence was found for increased bone resorption. Compared with children with osteogenesis imperfecta (OI), IJO patients had a similarly decreased cancellous BV but a much lower bone turnover. These results suggest a pathogenetic model for IJO, in which impaired osteoblast team performance decreases the ability of cancellous bone to adapt to the increasing mechanical needs during growth. This will finally result in load failure at sites where cancellous bone is essential for stability. PMID- 10804028 TI - Lactam formation increases receptor binding, adenylyl cyclase stimulation and bone growth stimulation by human parathyroid hormone (hPTH)(1-28)NH2. AB - Human parathyroid hormone (1-28)NH2 [hPTH(1-28)NH2] is the smallest of the PTH fragments that can fully stimulate adenylyl cyclase in ROS 17/2 rat osteoblast like osteosarcoma cells. This fragment has an IC50 of 110 nM for displacing 125I [Nle8,18,Tyr34]bovine PTH(1-34)NH2 from HKRK B7 porcine kidney cells, which stably express 950,000 human type 1 PTH/PTH-related protein (PTHrP) receptors (PTH1Rs) per cell. It also has an EC50 of 23.9 nM for stimulating adenylyl cyclase in ROS 17/2 cells. Increasing the amphiphilicity of the alpha-helix in the residue 17-28 region by replacing Lys27 with Leu and stabilizing the helix by forming a lactam between Glu22 and Lys26 to produce the [Leu27]cyclo(Glu22 Lys26)hPTH(1-28)NH2 analog dramatically reduced the IC50 for displacing 125I [Nle8,18,Tyr34]bPTH(1-34)NH2 from hPTH1Rs from 110 to 6 nM and dropped the EC50 for adenylyl cyclase stimulation in ROS 17/2 cells from 23.9 to 9.6 nM. These modifications also increased the osteogenic potency of hPTH(1-28)NH2. Thus, hPTH(1-28)NH2 did not significantly stimulate either femoral or vertebral trabecular bone growth in rats when injected daily at a dose of 5 nmol/100 g body weight for 6 weeks, beginning 2 weeks after ovariectomy (OVX), but it strongly stimulated the growth of trabeculae in the cancellous bone of the distal femurs and L5 vertebrae when injected at 25 nmol/100 g body weight. By contrast [Leu27]cyclo(Glu22-Lys26)hPTH(1-28)NH2 significantly stimulated trabecular bone growth when injected at 5 nmol/100 g of body weight. Thus, these modifications have brought the bone anabolic potency of hPTH(1-28)NH2 considerably closer to the potencies of the larger PTH peptides and analogs. PMID- 10804029 TI - The intracellular target for the antiresorptive aminobisphosphonate drugs in Dictyostelium discoideum is the enzyme farnesyl diphosphate synthase. AB - Aminobisphosphonate (aBP) drugs inhibit osteoclast-mediated bone resorption and also growth of amoebas of Dictyostelium discoideum apparently by interaction with the same intracellular target. Identification of the target in Dictyostelium therefore could also identify the target in osteoclasts. The aBPs (100 microM alendronate and 30 microM YM-175) inhibited conversion of [14C]mevalonate into sterols by cultures of Dictyostelium amoebas. One of three enzymes (isopentenyl diphosphate [IDP] isomerase, farnesyl diphosphate [FDP] synthase, and squalene synthase) appeared to be the target for this inhibition because conversion of [14C]IDP into squalene, the immediate precursor for sterol biosynthesis, was inhibited in extracts of wild-type amoebas by alendronate (IC50 = 75 nM) or risedronate (IC50 = 30 nM) whereas, when the extract had been prepared from amoebas of strains selected for having partial resistance to the growth inhibitory effects of alendronate (strain MR102) or risedronate (strain RB101), the values of IC50 were increased to 700 nM for alendronate (MR102 extract) or 130 nM for risedronate (RB101 extract). Neither IDP isomerase nor squalene synthase was inhibited significantly by alendronate or risedronate but both of these aBP drugs, and all others tested, inhibited FDP synthase. Determination of the nucleotide sequences of complementary DNAs (cDNAs) encoding FDP synthase in the wild-type and aBP-resistant strains of Dictyostelium indicated that there had been no changes in the amino acid sequence of the enzyme in the mutant strains. However, both mutant strains overproduce FDP synthase. It is concluded that FDP synthase is the intracellular target for the aBP drugs. PMID- 10804030 TI - Increased distraction rates influence precursor tissue composition without affecting bone regeneration. AB - The effect of increased distraction rate on bony tissue differentiation was studied using a paired bilateral model of rat femur lengthening. After a 6-day latency period, one randomly selected femur for each rat was distracted at 0.5 mm/day (normal rate) for 12 days, and the contralateral femur was distracted at 1.5 mm/day (increased rate) for 4 days. Femoral lengthening for each side was 6.0 mm, leaving the increased rate leg with an extra 8 days of consolidation compared with the normal rate limb. Group I rats (n = 9) were killed at day 18 postsurgery and analyzed for cartilage tissue composition and distribution. Group II rats (n = 7) were killed on day 36 postsurgery and analyzed by three-dimensional microcomputed tomography (MCT) for changes in new bone volume. Digital color analysis of slides stained with type II collagen antibody showed increases in cartilaginous tissue formation on the increased rate side (1.51 mm2 vs. 0.83 mm2; p = 0.10). No differences in new bone volume were detected between increased rate limbs and their contralateral controls (46.13 mm3 vs. 42.69 mm3; p = 0.63). These findings suggest that intermediate distraction rates may influence precursor tissue composition without affecting the final amount of new bone formed. Because damage to the tissue was not detected at either time point, these changes in chondrogenesis may reflect sensitivity of the pluripotential gap tissue to tension accumulation during lengthening. Future work with this in vivo model is focused on improving our understanding of the mechanisms behind this strain sensitivity. PMID- 10804031 TI - Resurgent malaria at the millennium: control strategies in crisis. AB - Completion of the Panama Canal in 1914 marked the beginning of an era of vector control that achieved conspicuous success against malaria. In 1955 the World Health Organization (WHO) adopted the controversial Global Eradication Campaign emphasising DDT (dichlorodiphenyltrichloroethane) spraying in homes. The incidence of malaria fell sharply where the programme was implemented, but the strategy was not applied in holoendemic Africa. This, along with the failure to achieve eradication in larger tropical regions, contributed to disillusionment with the policy. The World Health Assembly abandoned the eradication strategy in 1969. A resurgence of malaria began at about that time and today reaches into areas where eradication or control had been achieved. A global malaria crisis looms. In 1993 the WHO adopted a Global Malaria Control Strategy that placed priority in control of disease rather than infection. This formalises a policy that emphasises diagnosis and treatment in a primary healthcare setting, while de emphasising spraying of residual insecticides. The new policy explicitly stresses malaria in Africa, but expresses the intent to bring control programmes around the world into line with the strategy. This review raises the argument that a global control strategy conceived to address the extraordinary malaria situation in Africa may not be suitable elsewhere. The basis of argument lies in the accomplishments of the Global Eradication Campaign viewed in an historical and geographical context. Resurgent malaria accompanying declining vector control activities in Asia and the Americas suggests that the abandonment of residual spraying may be premature given the tools now at hand. The inadequacy of vector control as the primary instrument of malaria control in holoendemic Africa does not preclude its utility in Asia and the Americas. PMID- 10804032 TI - Role of tumour necrosis factor-alpha in the progression of heart failure: therapeutic implications. AB - The experimental and clinical evidence that demonstrates the effect of various cytokines, and in particular tumour necrosis factor (TNF)alpha, in patients with heart failure continues to accumulate. It is well established that increased levels of TNFalpha appear in the circulation of patients with heart failure and that the levels may have prognostic significance. Also, increased circulating TNFalpha levels may be responsible for the decreased expression of myocardial TNF receptors observed in failing myocardium. Along with these clinical data, it has been clearly demonstrated that increased levels of TNFalpha lead to cardiomyopathy and eventually death in experimental animals. Therefore, it is reasonable to assume that the increased levels of TNFalpha in patients with heart failure may be detrimental to cardiac function. The hypothesis that TNFalpha contributes to the pathogenesis of heart failure has recently been tested at the clinical level. The results of specific TNFalpha antagonism in patients with symptomatic heart failure demonstrate that anti-TNFalpha therapy is well tolerated and may be effective. This hypothesis is currently being tested in a large randomised, multicentre study that is expected to be complete within the next 2 years. Perhaps the most important aspect of the evolving research into the role of cytokines in heart failure is that the recognition of activation of inflammatory mediators provides new targets for therapeutic intervention. PMID- 10804034 TI - Systemic enzyme therapy in oncology: effect and mode of action. AB - Plant extracts with a high content of proteolytic enzymes have been used for a long time in traditional medicine. Besides proteolytic enzymes from plants, 'modern' enzyme therapy additionally includes pancreatic enzymes. The therapeutic use of proteolytic enzymes is partly based on scientific studies and is partly empirical. The aim of the current review is to provide an overview of clinical trials of systemic enzyme therapy in oncology, and to discuss the evidence for their possible mechanisms of action. Clinical studies of the use of proteolytic enzymes in oncology have mostly been carried out on an enzyme preparation consisting of a combination of papain, trypsin and chymotrypsin. This review of these studies showed that enzyme therapy can reduce the adverse effects caused by radiotherapy and chemotherapy. There is also evidence that, in some types of tumours, survival may be prolonged. The beneficial effect of systemic enzyme therapy seems to be based on its anti-inflammatory potential. However, the precise mechanism of action of systemic enzyme therapy remains unsolved. The ratio of proteinases to antiproteinases, which is increasingly being used as a prognostic marker in oncology, appears to be influenced by the oral administration of proteolytic enzymes, probably via an induction of the synthesis of antiproteinases. Furthermore, there are numerous alterations of cytokine composition during therapy with orally administered enzymes, which might be an indication of the efficacy of enzyme therapy. Effects on adhesion molecules and on antioxidative metabolism are also reviewed. PMID- 10804033 TI - Tyrosine kinase inhibitors targeted to the epidermal growth factor receptor subfamily: role as anticancer agents. AB - Abnormal cell signal transduction arising from protein tyrosine kinases has been implicated in the initiation and progression of a variety of human cancers. Over the past 2 decades pharmaceutical and university laboratories have been involved in a tremendous effort to develop compounds that can selectively modulate these abnormal signalling pathways. Targeting receptor tyrosine kinases, especially the epidermal growth factor receptor subfamily, has been at the forefront of this effort as a result of strong clinical data correlating over-expression of these receptors with more aggressive cancers. There are a variety of strategies under development for inhibiting the kinase activity of these receptors, targeting both the extracellular and intracellular domains. Antibody-based approaches, immunotoxins and ligand-binding cytotoxic agents use the extracellular domain for targeted tumour therapy. Small molecule inhibitors target the intracellular catalytic region by interfering with ATP binding, while nonphosphorylatable peptides are aimed at the intracellular substrate binding region. Compounds that inhibit subsequent downstream signals from the receptor by interrupting intracellular protein recognition sequences are also being investigated. In the past 5 years enormous progress has been made in developing tyrosine kinase inhibitor compounds with sufficient potency, bioavailability and selectivity against this subfamily of receptor tyrosine kinases. The anti-HER2 monoclonal antibody, trastuzumab, for patients with metastatic breast cancer is the first of these inhibitor compounds to gain FDA approval. However, preclinical and clinical trials are ongoing with a variety of other monoclonal antibodies, immunotoxins, and small molecule quinazoline and pyrimidine-based inhibitors. Although their cytotoxic and cytostatic potential has been proven, they are not likely to replace standard chemotherapy regimens as single-agent, first-line therapeutics. Instead, their promising additive and synergistic antitumour effects in combination with standard chemotherapeutics suggest that these novel agents will find their greatest utility and efficacy in conjunction with existing anticancer agents. PMID- 10804035 TI - A risk-benefit assessment of pharmacological and nonpharmacological treatments for nausea and vomiting of pregnancy. AB - Despite evidence of fetal safety, most antiemetics are contraindicated in pregnancy. We summarise a risk-benefit analysis of the literature on safety and effectiveness of pharmacotherapy and nontraditional therapy for nausea and vomiting of pregnancy (NVP) to provide evidence-based guidelines on the management of NVP. The medical literature was scanned for controlled studies on the human teratogenicity and effect of various antiemetics in pregnant women. Data were pooled based on drug/therapy class and summarised to determine relative risk with 95% confidence interval (for malformations and failure rates for NVP) and homogeneity (chi-square test). Evidence from controlled trials has demonstrated the safety and efficacy of the following drugs for the treatment of varying degrees of NVP: doxylamine/pyridoxine+/-dicycloverine (dicyclomine), antihistamine H1 receptor antagonists, and phenothiazines (as a group). However, pooled data for doxylamine/pyridoxine+/-dicycloverine, H1 antagonists and phenothiazines were not homogeneous. Other therapies, such as pyridoxine alone, metoclopramide, ondansetron and the corticosteroids may be beneficial in managing NVP. However, limited efficacy studies and the paucity of well-controlled safety studies may limit the use of some of these agents among patients not responsive to first-line agents. Well-controlled safety and effectiveness trials in patients with NVP are lacking for nonpharmacological treatments (e.g. acupressure). NVP can be managed safely and effectively. Further trials must be conducted in order to determine the true effectiveness of certain agents in patients with NVP. PMID- 10804036 TI - Concurrent gonococcal and chlamydial infection: how best to treat. AB - Clinicians treating concurrent gonococcal and chlamydial infections have a variety of drugs to choose from. Neisseria gonorrhoeae is adept at developing resistance and the choice of antibiotic must be dictated to some extent by the patterns of resistance in the locality of the clinician. In contrast, resistance of Chlamydia trachomatis to some classes of drugs has been shown in vitro but does not appear to be clinically important at present. The success of treatment depends on patient compliance with the drug administration schedule. With these organisms, which can be carried asymptomatically, many patients are unlikely to comply with courses of antibiotics. Although single-dose therapy with azithromycin is available and established for chlamydial genital infection, it is more expensive and difficult to justify in a cash limited Healthcare system, and its efficacy for treating concurrent gonococcal infection requires further study. In patients where compliance is likely to be of concern, its use may be justified. Another major deterrent for completing antibiotic courses is the adverse effect profile. Most of the available drugs cause only minor adverse effects, in particular gastrointestinal. Ofloxacin has a better profile than doxycycline but is considerably more expensive. Newer fluoroquinolones, found to be effective in vitro, are being assessed in clinical studies. However, more evidence is required before recommending these over the tried and tested therapies. PMID- 10804037 TI - Linezolid. AB - Linezolid is an oxazolidinone antibacterial agent that acts by inhibiting the initiation of bacterial protein synthesis. Cross-resistance between linezolid and other inhibitors of protein synthesis has not been demonstrated. Linezolid has a wide spectrum of activity against gram-positive organisms including methicillin resistant staphylococci, penicillin-resistant pneumococci and vancomycin resistant Enterococcus faecalis and E. faecium. Anerobes such as Clostridium spp., Peptostreptococcus spp. and Prevotella spp. are also susceptible to linezolid. Linezolid is bacteriostatic against most susceptible organisms but displays bactericidal activity against some strains of pneumococci, Bacteroides fragilis and C. perfringens. In clinical trials involving hospitalised patients with skin/soft tissue infections (predominantly S. aureus), intravenous/oral linezolid (up to 1250 mg mg/day) produced clinical success in >83% of individuals. In patients with community-acquired pneumonia, success rates were >94%. Preliminary clinical data also indicate that twice daily intravenous/oral linezolid 600 mg is as effective as intravenous vancomycin 1 g in the treatment of patients with hospital-acquired pneumonia and in those with infections caused by methicillin-resistant staphylococci. Moreover, linezolid 600 mg twice daily produced >85% clinical/microbiological cure in vancomycin-resistant enterococcal infections. Linezolid is generally well tolerated and gastrointestinal disturbances are the most commonly occurring adverse events. No clinical evidence of adverse reactions as a result of monoamine oxidase inhibition has been reported. PMID- 10804038 TI - Racecadotril. AB - Racecadotril is an oral enkephalinase inhibitor used in the treatment of acute diarrhoea. It prevents the degradation of endogenous opioids (enkephalins), thereby reducing hypersecretion of water and electrolytes into the intestinal lumen. In a randomised double-blind study in 6 adult volunteers with castor oil induced diarrhoea, racecadotril significantly reduced stool weight and stool number in comparison with placebo. Similar results have been obtained in treating castor oil-induced diarrhoea in rats. Racecadotril was significantly more effective than placebo in randomised double-blind studies in adults or children with diarrhoea (of infectious origin or in adults with HIV infection). In well controlled trials, racecadotril had efficacy similar to that of loperamide and was generally as effective as loperamide-oxide. Racecadotril had a similar tolerability profile to placebo, and was better tolerated than loperamide, in adults and children with diarrhoea. It caused significantly less constipation after resolution of diarrhoea than loperamide. PMID- 10804039 TI - Valaciclovir: a review of its long term utility in the management of genital herpes simplex virus and cytomegalovirus infections. AB - Valaciclovir is an aciclovir prodrug used to treat infections caused by herpes simplex virus (HSV) and varicella zoster virus, and for prophylaxis against cytomegalovirus (CMV). Oral valaciclovir provides significantly better oral bioavailability than oral aciclovir itself, contributing to the need for less frequent administration. Several studies have demonstrated the efficacy of long term (> 90 days) therapy with valaciclovir for the suppression of genital HSV disease in otherwise healthy individuals with HSV infection. In 1 randomised, double-blind trial, once daily valaciclovir (1000 mg, 500 mg and 250 mg) produced statistically significant suppression of disease recurrence, as did twice daily valaciclovir 250 mg and aciclovir 400 mg. Valaciclovir dosages of > or = 500 mg daily are recommended for suppression of genital herpes recurrences in immunocompetent individuals. This disease occurs frequently in patients with human immunodeficiency virus (HIV) infection and, in a single randomised double blind trial, prophylactic valaciclovir (1000 mg once daily or 500 mg twice daily) and aciclovir (400 mg twice daily) were found to be of similar efficacy in the suppression of genital herpes. However, a higher than expected dropout rate indicated that more studies of valaciclovir in patients with HIV are required. In a randomised trial of patients undergoing renal transplant, valaciclovir 2 g 4 times daily for 90 days significantly reduced the incidence and delayed the onset of CMV disease: the incidence in valaciclovir-treated patients who were CMV seronegative at baseline, and recieived a kidney from a CMV-seropositive donor, was 3% versus 45% for placebo after 90 days of treatment. Acute graft rejection was also reduced in the valaciclovir-treated group. A small study in heart transplant patients compared valaciclovir (2 g 4 times daily) with aciclovir (200 mg 4 times daily) and found a significant reduction in CMV antigenaemia favouring valacilovir at the end of the treatment period. Additional reductions in other indices of CMV in those given valaciclovir compared with aciclovir were also noted. In a preliminary study of prophylaxis for CMV disease in bone marrow transplant recipients valaciclovir (2 g 4 times daily) was superior to aciclovir (800 mg 4 times daily) in terms of time to CMV viraemia or viruria. Although valaciclovir (8 g/day for approximately 30 weeks) reduced the incidence and time to CMV disease compared with aciclovir (3.2 g/day) in patients with advanced HIV disease, valaciclovir was associated with more gastrointestinal complaints and an increased risk of death, leading to premature termination of the study. As yet, no trials comparing the efficacy of valaciclovir with famciclovir (the oral prodrug for penciclovir) in the suppression of recurrent episodes of genital herpes have been published, nor have direct comparisons been made, between valaciclovir with ganciclovir in patients with CMV disease. Valaciclovir is well tolerated at dosages used to suppress recurrent episodes of genital herpes (500 to 1000 mg/day) in immunocompetent and HIV seropositive individuals, with headache being reported most often. However, a potentially fatal thrombotic microangiopathy (TMA)-like syndrome has been reported in some immunocompromised patients receiving high-dose prophylactic valaciclovir therapy (8 g/day) for CMV disease for prolonged periods, and the risk of this syndrome appears to be higher in patients with advanced HIV disease. While the clinical benefits of valaciclovir in some immunocompromised patients may outweigh the risk of TMA, close monitoring for symptoms of TMA is indicated in all immunocompromised patients receiving high-dose valaciclovir. CONCLUSION: Oral valaciclovir is an effective drug for the suppression of recurrent episodes of genital herpes in immunocompetent and immunocompromised individuals. (ABSTRACT TRUNCATED) PMID- 10804040 TI - Zolpidem: an update of its pharmacology, therapeutic efficacy and tolerability in the treatment of insomnia. AB - Zolpidem is an imidazopyridine agent that is indicated for the short term (< or = 4 weeks) treatment of insomnia (recommended dosage 10 mg/day in adults and 5 or 10 mg/day in the elderly or patients with hepatic impairment). Data have shown that the hypnotic efficacy of zolpidem is generally comparable to that of the benzodiazepines flunitrazepam, flurazepam, nitrazepam, temazepam and triazolam as well as nonbenzodiazepine hypnotic agents such as zopiclone and trazodone in the treatment of elderly and adult patients with insomnia. The comparative efficacy of a recently available nonbenzodiazepine hypnotic zaleplon and zolpidem has yet to be established. There was no evidence of tolerance developing to the hypnotic effects of zolpidem in a number of studies of up to 6 months' duration. However, tolerance has been described in a few patients taking the drug at high dosages for periods of up to several years. Zolpidem is well tolerated in patients with insomnia and the most common adverse events are generally nausea, dizziness and drowsiness. Although zolpidem produced some psychomotor and memory impairment over the first few hours after administration, it had few next-day effects (including effects on daytime well-being and morning coordination). In this respect, it was comparable or superior to flunitrazepam and flurazepam and comparable to other benzodiazepines in patients with insomnia. Zolpidem appears to have a low potential for abuse. CONCLUSIONS: Zolpidem is effective and well tolerated in patients with insomnia, including the elderly. Studies have shown that zolpidem generally has similar efficacy to other hypnotics including benzodiazepines and zopiclone. Zolpidem appears to have minimal next-day effects on cognition and psychomotor performance when administered at bedtime. In addition, there is little evidence of tolerance to the hypnotic effects of zolpidem, or rebound insomnia or withdrawal symptoms after discontinuation of the drug when it is given as recommended (10 mg/day for < 1 month) or over longer periods. PMID- 10804041 TI - Montelukast: a review of its therapeutic potential in persistent asthma. AB - Montelukast is a cysteinyl leukotriene receptor antagonist used to treat persistent asthma in patients aged > or = 6 years. The drug has a rapid onset of action. Improvements in lung function and reductions in as-needed beta2-agonist usage are apparent within 1 day of initiating montelukast treatment in adults and adolescents (aged > or = 15 years treated with 10 mg/day) or children (aged 6 to 14 years treated with 5 mg/day) with persistent asthma as shown in clinical trials. In two 12-week, multicentre, randomised, double-blind studies in adults and adolescents aged > or = 15 years with persistent asthma [forced expiratory volume in 1 second (FEV1) = 50 to 85% predicted] there was significantly (p < 0.05) greater improvement in FEV1, symptom scores, peak expiratory flow (PEF), as needed beta2-agonist use, peripheral eosinophil counts and health-related quality of life (QOL) in patients treated with montelukast 10 mg/day than in recipients of placebo. Improvements were significantly greater in patients treated with inhaled beclomethasone 400 microg/day than in recipients of montelukast 10 mg/day in 1 of these studies. Nonetheless, 42% of montelukast recipients experienced > or = 11% improvement in FEV1, the median improvement in this parameter in beclomethasone-treated patients. In an 8-week multicentre, randomised, double blind, study in children aged 6 to 14 years with persistent asthma (FEV1 50 to 85% predicted), montelukast 5 mg/day produced significantly greater improvements in FEV1, clinic PEF, as-needed beta2-agonist use, peripheral eosinophil counts, asthma exacerbations and QOL scores than placebo. The combination of montelukast 10 mg/day plus inhaled beclomethasone 200 microg twice daily provided significantly better asthma control than inhaled beclomethasone 200 microg twice daily in adults with poorly controlled asthma (mean FEV1 = 72% predicted) despite 4 weeks treatment with inhaled beclomethasone. Patients receiving the combination experienced significant improvements in FEV1 and morning PEF, significant reductions in daytime symptom scores, as-needed beta2 agonist usage and night time awakenings with asthma, and had significantly lower peripheral blood eosinophil counts after 16 weeks in this multicentre, randomised, double-blind, placebo-controlled study. Among adults (FEV1 > or = 70%) treated with montelukast 10 mg/day for 12 weeks, inhaled corticosteroid dosages were titrated downward by 47% (vs 30% in placebo recipients), 40% of patients were tapered off of inhaled corticosteroids (vs 29%), and significantly fewer patients (16 vs 30%) experienced failed corticosteroid rescues in a multicentre, randomised, double blind study. During clinical studies, the frequency of adverse events in montelukast-treated adults, adolescents and children was similar to that in placebo recipients. In conclusion, montelukast is well tolerated and effective in adults and children aged > or = 6 years with persistent asthma including those with exercise-induced bronchoconstriction and/or aspirin sensitivity. Furthermore, montelukast has glucocorticoid sparing properties. Hence, montelukast, as monotherapy in patients with mild persistent asthma, or as an adjunct to inhaled corticosteroids is useful across a broad spectrum of patients with persistent asthma. PMID- 10804042 TI - Prolonged-release mesalazine: a review of its therapeutic potential in ulcerative colitis and Crohn's disease. AB - Prolonged-release mesalazine (Pentasa) consists of ethylcellulose-coated microgranules from which mesalazine (known in the US as mesalamine) is released in the small and large intestine in a diffusion-dependent manner. Dose-dependent improvements in clinical and endoscopic parameters have been reported with prolonged-release mesalazine 2 and 4 g/day in clinical trials in patients with mild to moderately active ulcerative colitis. Induction of clinical and endoscopic remission was achieved in more patients receiving a daily dosage of 4 g/day than in those receiving placebo. In patients with ulcerative colitis in remission, prolonged-release mesalazine is effective in reducing the rate of relapse. Higher dosages tend to be more effective, and a 12-month remission rate of 64% has been reported for patients treated with a 4 g daily dosage of this formulation. Comparative data indicate that prolonged-release mesalazine has similar efficacy in maintaining remission to molar equivalent doses of sulfasalazine. Data from a study in patients with mild to moderately active Crohn's disease indicates that higher dosages (4 g/day) of prolonged-release mesalazine are more effective than placebo in reducing disease activity. After 16 weeks' treatment, 64% of patients receiving a 4 g/day dosage experienced clinical improvement and 43% attained remission. In studies of patients in remission of Crohn's disease, the formulation appears to be more effective in preventing relapse in patients with isolated small bowel disease than in those with colonic involvement. The tolerability profile of oral prolonged-release mesalazine is similar to that of placebo and the incidence of adverse events does not appear to be dose-related. Nausea/vomiting, diarrhoea, abdominal pain and dyspepsia occur most frequently, although their incidence is low. Reports of nephrotoxicity during prolonged-release mesalazine treatment are rare. CONCLUSIONS: Oral prolonged-release mesalazine is effective for maintenance and induction of remission of mild to moderately active colitis, both in patients with distal disease and in those with pancolitis. The formulation has similar efficacy to that of equimolar concentrations of sulfasalazine. Prolonged-release mesalazine also appears to be effective in the treatment of Crohn's disease, and maintenance therapy is of particular value in patients with isolated small bowel involvement. Evidence suggests that higher dosages (3 to 4 g/day) of prolonged-release mesalazine have additional therapeutic benefits over lower dosages in patients with inflammatory bowel disease without increasing the incidence of adverse events. PMID- 10804043 TI - Celecoxib: a review of its use in osteoarthritis, rheumatoid arthritis and acute pain. AB - Celecoxib is a cyclo-oxygenase (COX) inhibitor that exhibits relative in vitro and ex vivo selectivity for COX-2 over COX-1. Results of randomised double-blind multicentre studies indicate that celecoxib is superior to placebo and has similar efficacy as conventional nonsteroidal anti-inflammatory drugs (NSAIDs) in improving the signs and symptoms of osteoarthritis and rheumatoid arthritis. Analgesic efficacy and improvements in functional status are apparent within 2 weeks of starting therapy and are maintained throughout treatment. Available data suggest that celecoxib has analgesic efficacy in patients with postsurgical dental pain, although this is yet to be confirmed. In patients with osteoarthritis of the knee, celecoxib 100 and 200 mg and naproxen 500 mg twice daily were similarly efficacious and superior to placebo. Once and twice daily celecoxib dosage regimens provided comparable efficacy. Improvements in physical function paralleled those in pain relief. Celecoxib also has efficacy in treating the signs and symptoms of osteoarthritis of the hip. The effects of celecoxib were not diminished in elderly patients with osteoarthritis of the hip or knee. All dosages of celecoxib (100 to 400 mg twice daily) and naproxen 500 mg twice daily produced significant anti-inflammatory and analgesic effects in patients with active rheumatoid arthritis. In patients with stable rheumatoid arthritis, celecoxib 200 mg twice daily showed sustained symptomatic improvements similar to those of twice daily slow-release diclofenac 75 mg over a 24-week period. Celecoxib was well tolerated in clinical trials. Upper gastrointestinal complications occurred in significantly fewer patients treated with twice daily celecoxib 25 to 400 mg than in those receiving comparator NSAIDs. There was no evidence of a dose relationship in endoscopic ulcer development and incidences in celecoxib and placebo recipients were lower than in those receiving twice daily naproxen 500 mg or ibuprofen 800 mg 3 times daily. CONCLUSIONS: Celecoxib is the first COX-2 specific inhibitor approved for use in osteoarthritis and rheumatoid arthritis. Celecoxib produces significant improvements in pain and inflammation and these effects are maintained during treatment for up 24 weeks in clinical trials. Studies indicate that celecoxib has similar efficacy to conventional NSAIDs in relieving pain and improving functional status, but is associated with a lower incidence of upper gastrointestinal ulceration and complications. This promising gastrointestinal safety profile, together with sustained symptomatic relief, places celecoxib as a useful alternative for the treatment of osteoarthritis and rheumatoid arthritis, particularly in patients at high risk of developing gastrointestinal events. Although data are encouraging, its place in acute pain states remains to be established. PMID- 10804046 TI - The occipital transtentorial approach for cerebellar arteriovenous malformation in a child. AB - This report presents a case of arteriovenous malformation (AVM) in the region of the superior vermis and quadrigeminal plate, which was partially embolized using a endovascular technique and subsequently surgically excised. An occipital transtentorial approach was employed to excise the AVM, and the patient did well without neurological complications postoperatively. Comparing various surgical approaches, we believe the occipital transtentorial approach is the best for the AVM of this region. PMID- 10804044 TI - Ebastine: an update of its use in allergic disorders. AB - Ebastine is a second-generation antihistamine which undergoes transformation to its active metabolite, carebastine. Its antihistaminic and antiallergic effects have been demonstrated in in vitro and in vivo studies, in addition to data obtained from clinical trials. Patients with allergic rhinitis or chronic idiopathic urticaria experienced significant improvement in their symptoms with ebastine 10 or 20 mg once daily. Some studies in patients with seasonal allergic rhinitis (SAR) have indicated trends towards greater efficacy with the 20 mg than the 10 mg dose, although only 1 study has shown statistically significant benefits. In comparative trials in patients with SAR, ebastine 10 mg was as effective as most other second-generation antihistamines, including astemizole, azelastine, cetirizine, loratadine and terfenadine. Ebastine 20 mg/day was significantly superior to loratadine 10 mg/day in patients with SAR according to effects on secondary efficacy variables in comparative studies; 1 study found significantly greater changes from baseline in mean total symptom score with ebastine 20 mg (-43 vs -36% with loratadine, p = 0.045). In patients with perennial allergic rhinitis, ebastine 10 or 20 mg daily was significantly more effective than loratadine in reducing total symptom scores from baseline 1 comparative study. There have been no reports of serious adverse cardiac effects during ebastine therapy. Increases in corrected QT interval have been observed during clinical trials; however, these have not been considered clinically significant and were generally of similar magnitude to those seen with loratadine. The normal diurnal variation in QTc interval and the problems associated in correcting for changes in heart rate also complicate assessment of this issue. The incidence of adverse events during ebastine treatment is not significantly greater than that observed with placebo or other second-generation antihistamines. CONCLUSIONS: Ebastine 10 mg daily is a well tolerated and effective treatment for allergic rhinitis and chronic idiopathic urticaria. At this dosage, it is as effective as the other second-generation antihistamines against which it has been compared. Ebastine 20 mg has similar tolerability to the 10 mg dose, and trends towards greater efficacy with the higher dose have been shown in some studies. Ebastine does not appear to be associated with any significant cardiac adverse events. Ebastine is a useful treatment option for patients with allergic rhinitis or chronic idiopathic urticaria. PMID- 10804045 TI - Bupropion: a review of its use in the management of smoking cessation. AB - Sustained release bupropion (amfebutamone) is a non-nicotine agent that is indicated as an aid to smoking cessation. In 2 large well designed clinical trials, sustained release bupropion 300 mg/day (the recommended dose) for 7 or 9 weeks was associated with considerably and significantly higher smoking abstinence rates (continuous abstinence and 7-day point prevalence rates) than placebo during treatment and at follow-up at 6 and 12 months. Point prevalence rates at 12 months in 2 studies were 23.1 and 30.3% with bupropion, whereas values for placebo were 12.4 and 15.6%. Continuous abstinence rates at 12 months, available from 1 trial, were 18.4% with bupropion and 5.6% with placebo. Furthermore, bupropion was associated with significantly higher quitting rates than nicotine patch in a comparative study. Combination therapy with bupropion and nicotine patch provided slightly higher abstinence rates than bupropion alone, although differences were not statistically significant. The combination was superior to nicotine patch alone. Data from a preliminary report of long term bupropion treatment (52 weeks) showed that the drug was associated with significantly higher continuous abstinence rates than placebo only to 6 months. However, point prevalence abstinence rates were significantly higher with bupropion than placebo to 18 months. Bupropion 300 mg/day recipients reported nicotine withdrawal symptoms during treatment; however, the symptoms were significantly less severe with bupropion than placebo. Patients receiving bupropion 300 mg/day or bupropion in combination with nicotine patch for smoking cessation generally gained less bodyweight than placebo recipients. The benefits of bupropion for preventing weight gain persisted after the completion of long term, but not short term therapy. Bupropion was well tolerated in clinical trials, and the only adverse events that were significantly more common with bupropion than placebo were insomnia and dry mouth. Data published so far suggest that sustained release bupropion has a low potential for inducing seizures (seizure rate approximately 0.1% in patients with depression). CONCLUSIONS: Bupropion is an effective and well tolerated smoking cessation intervention. Further studies with long term follow-up will be useful in determining whether abstinence rates are maintained with bupropion. In addition, clarification of its efficacy in comparison with other therapies used for smoking cessation would help to establish its clinical value. The reduced potential for weight gain with bupropion and the ability to use bupropion in combination with nicotine replacement therapy make the drug a useful treatment option for smoking cessation. PMID- 10804047 TI - Pediatric acoustic schwannoma showing rapid regrowth with high proliferative activity. AB - Acoustic schwannoma is a slow-growing tumor and usually occurs in adult patients. We report a rare pediatric case of acoustic schwannoma with high proliferative potential. A 10-year-old boy was diagnosed as having a right cerebellopontine angle tumor. The tumor was subtotally resected. Histological examination revealed a typical acoustic schwannoma with a few mitotic figures. Chromosomal analysis showed no abnormality on the long arm of chromosome 22 associated with neurofibromatosis type 2. The lesion re-grew rapidly as an acoustic schwannoma, necessitating subtotal resection on three occasions and CyberKnife radiosurgery. The immunohistochemical MIB-1 staining indices of the specimens obtained at the first, second, and third operations were 2.3%, 4.6% and 14.7%, respectively. The immunohistochemical proliferative potential of acoustic schwannoma is discussed. PMID- 10804048 TI - Giant cystic craniopharyngiomas with extension into the posterior fossa. AB - Between 1991 and 1998, 24 patients underwent surgery for a craniopharyngioma in our department. This group included two patients who had tumors with extensive growth along the midline, and along the posterior fossa in particular. In both cases suprasellar calcifications were typical features on magnetic resonance imaging (MRI) and computed tomography (CT). A 7-year-old boy presented with a 6 month history of headache, nausea, and progressive unilateral hearing loss. With a suboccipital approach it was possible to remove the main part of the tumor. In a 13-year-old boy headache and visual deterioration led to the diagnosis of a craniopharyngioma, which was removed with a pterional approach. For the neuroimaging work-up in such cases of atypically growing craniopharyngiomas MRI is the method of choice. Additional CT scanning is recommended, which provides valuable information about bony changes at the skull base due to space-occupying growth. CT substantiates the differential diagnosis if typical calcifications are seen. PMID- 10804049 TI - Factors in neurological deterioration and role of surgical treatment in lumbosacral spinal lipoma. AB - The purpose of this study was to determine factors that might be involved in neurological deterioration and the role of surgical treatment in patients with lumbosacral spinal lipoma. Pre- and postoperative courses of 34 patients were retrospectively analyzed. The age at surgery ranged from 1 month to 47 years. The records of preoperative neurological status indicated that older patients had more severe deficits, while all 8 asymptomatic patients were under 5 years of age. Motor deficits were noted in 9 patients, in 7 of whom the lipoma extended cranially beyond the L5 level. Transitional-type lipomas were accompanied by more severe deficits (asymptomatic 1, symptomatic 17) than other types (asymptomatic 7, symptomatic 9). Postoperative follow-up periods ranged from 5 months to 13 years. During these periods, 7 of the 8 asymptomatic patients remained neurologically intact. Nine of the 26 symptomatic patients improved. Age, extension of the lipoma in the spinal canal and type of lipoma will influence the preoperative neurological status of the patients. Early untethering surgery is recommended in patients with large lipomas extending beyond the L5 level. PMID- 10804051 TI - How should we manage children after mild head injury? AB - There are many controversies concerning the management of children after mild head injury. Most of these patients achieve a full recovery without medical or surgical intervention. A small percentage of them deteriorate owing to intracranial complications. The goal of this study was to identify significant factors that might allow the identification of patients at risk of subsequent deterioration. Its secondary goal was to establish a clinical protocol for the management of mild head injuries in children. We retrospectively reviewed the records of 166 children and adolescents with head trauma who had Glasgow Coma Scale (GCS) or Children Coma Scale (CCS) scores of 13-15 at the time of admission. The patients were divided into five age categories: babies younger than 1 year, children 1-3, 4-6, and 7-14 years old, and adolescents 15-17 years of age. The largest age group consisted of children 7-14 years old (83 cases). There was a male predominance (2:1). The main causes of injury were traffic accidents (55 cases) and falls (53 patients). Neurosurgical procedures were required in 93 of the 166 patients (56%). The most common intracranial lesion was subdural and epidural hematoma (60 cases). In 26 children (15.6%) diffuse brain swelling was the only lesion. A skull fracture was found in 103 cases and was accompanied by epidural hematoma (HED) in 19 cases (18%) and by subdural hematoma (HSD) in 12 cases (12%). However, the 63 children without a fracture also included 18 (29%) who had HSD and 11 (17%) who had HED. In our population 165 (99%) of the patients obtained a very good or good result. None was left severely disabled or in a vegetative state. One patient with GCS 13 died of an infection. We concluded that skull X-ray examination is not sufficient to rule out intracranial hematoma. We recommend CT scanning and admission to hospital for 24 h observation for all children with minor head injury, because of the risk of delayed hematoma. PMID- 10804050 TI - Patients with urinary incontinence often benefit from surgical detethering of tight filum terminale. AB - We retrospectively reviewed 77 patients with a tethered spinal cord syndrome to evaluate the results of neurosurgical treatment. The patients were divided into two groups: in group 1 there were 17 patients with primary tethered cord who had normal level conus medullaris (NLCM) and normal thickness filum terminale (NTFT) with urinary incontinence, and group 2 was made up of 60 patients with secondary spinal cord tethering after a previous closure of a midline fusion defect who had a low-lying conus medullaris. Neurological examination, radiography, urodynamic tests and electrophysiological findings confirmed the diagnosis. Conventionally, tethered cord syndrome has been defined as a state in which the conus medullaris is located below the L1-2 disc space. However, in a patient with urinary incontinence and a hyperreflexive type of neurogenic bladder, in whom the conus medullaris is found to be at the normal level, there may still be cord tethering that is causing the incontinence. In this study the comparison was based on evaluation of the response to treatment and general characteristics of the syndrome in both groups of patients to draw attention to the general approach to this incapacitating mal-development. PMID- 10804052 TI - Surgical outcome of pediatric hydrocephalus treated by endoscopic III ventriculostomy: prognostic factors and interpretation of postoperative neuroimaging. AB - In order to analyze the surgical outcome according to clinical characteristics and to evaluate the correlation between clinical improvement and neuroimaging changes, we retrospectively reviewed 32 children who had undergone endoscopic III ventriculostomy (ETV) from February 1994 to May 1998. There were 15 boys and 17 girls, with a mean age of 5.2 years (range: 1 month to 13 years). The etiology of the hydrocephalus was primary aqueductal stenosis in 18 patients, secondary aqueductal stenosis caused by tumors in 5, IV ventricle outlet obstruction in 5, and hydrocephalus associated with meningomyelocele in 4. The mean duration of follow-up was 19.4 months (range 1-50 months). Overall, surgical outcome was regarded as good in 21 of 29 patients. Surgical outcome was poor in patients younger than 1 year (P<0.05). Neuroimaging 1 month after ETV showed a decrease in ventricular size in 11 of the 16 patients with good surgical outcomes. Five showed minimal changes only. In patients with good outcomes, ventricular size tended to decrease as time passed. Resolution of periventricular edema, flow void in the III ventricle on T2-weighted axial images, and cine-MR imaging were sensitive indicators of good outcome. We suggest that ETV be considered as a primary treatment option in patients older than 1 year of age with noncommunicating hydrocephalus. In addition, time factors should be taken into consideration when surgical outcome is judged. Changes in ventricular size could not predict surgical outcome completely in themselves. Therefore, a comprehensive postoperative assessment should be made with the help of T2-weighted MRI and cine MRI. PMID- 10804053 TI - The importance of surgery in supratentorial ependymomas. Long-term survival in a series of 23 cases. AB - In view of the widely recognized correlation between extent of surgical resection and length of survival of children with intracranial ependymoma and the statement that total resection is more likely to be achieved in supratentorial than infratentorial primaries, we decided to review our experience with supratentorial ependymomas and the pertinent literature to verify the importance of surgery in treating this subgroup of pediatric ependymal neoplasms. Of 23 patients operated on, 12 are still alive without evidence of disease 72-357 months after surgery (mean 227, median 237 months). One girl treated by surgery alone was lost to follow-up after 234 months when she, and 7 other patients in the series, had already passed the end of the period of risk for recurrence according to Collins' law. Six surviving patients (2 with subependymoma and 4 with ependymoma) were treated by surgery alone and only 1, the oldest in the series, had to undergo a second operation for recurrence after 10 years. The idea of treating intracranial ependymoma by surgery alone was favored by eminent neurosurgeons in the past and has recently received renewed attention. This was in part the consequence of recognizing that unlike diffuse astrocytoma, in which neoplastic cells can be found up to several centimeters away from the apparent tumor borders, ependymoma has more or less well-defined margins and grows mainly by expansion. Early experience with the policy of electively deferring adjuvant therapy after radiologically controlled total resection of ependymoma seems encouraging, although postoperative MRI does not yet indicate absolute certainty. Close surveillance is recommended. The majority of ependymomas so far treated by surgery alone, with relatively good success, have been supratentorial. In conclusion, on the basis of our experience and a review of the literature we favor a change in attitude to the management of intracranial ependymomas, especially of the cerebrum, with radiologically controlled radical surgery alone followed by close surveillance with periodic MR imaging until the child passes the period of risk for recurrence according to Collins' law as the initial option. In children less than 3 years old the period of surveillance should be doubled. In case of recurrence, reoperation should be considered first, particularly for supratentorial primaries. Radiotherapy continues to be a major option in malignant ependymoma and unresectable primary or recurrent benign ependymoma. PMID- 10804054 TI - Cloverleaf skull and multiple congenital anomalies in a girl exposed to cocaine in utero: case report and review of the literature. AB - The case of a girl with cloverleaf skull (CLS) and multiple congenital anomalies is reported. Both parents have a history of drug use. Maternal cocaine abuse during the first trimester of pregnancy was obvious, and other drugs, such as marihuana and alcohol, were also taken by the mother. Many central nervous system malformations have been reported in association with cocaine abuse, the most severe being midline defects and neural tube defects. To our knowledge this is the first case reported of CLS anomaly associated with drug exposure. We also describe other anomalies not previously reported in association with CLS. PMID- 10804055 TI - Intraosseous neurinoma of the parietal bone. AB - Intraosseous neurinoma is very uncommon tumor. Its location in the skull is extremely rare. A 4-year-old boy presented with a lump in the right occipital region. Computed tomography revealed a soft tissue mass with bony erosion in the right occipital bone. The solid, nontender, and immobile mass was totally removed. The pathological examination showed that it was an intraosseous neurinoma. Total resection of intraosseous neurinomas is sufficient treatment and is not followed by recurrence. PMID- 10804056 TI - Transoral protrusion of a peritoneal catheter: a case report and literature review. AB - Transoral protrusion of a peritoneal catheter is rare. Only two cases have been reported in the English literature. We now report the case of a 5-year-old girl who presented with a catheter that had been inserted 4 years previously, protruding from her mouth. Signs of cerebrospinal fluid infection or peritonitis were absent. The peritoneal catheter was cut and externalized at the chest. The distal portion, which had perforated the stomach wall, was removed using endoscopic procedures. After 3 weeks of antibiotic treatment, a new shunt was inserted. Analysis of 50 cases of bowel perforation extracted from the English literature showed that among the suggested factors such as age, gender, nutritional state, history of abdominal surgery, and length and type of the catheter, age was the only predisposing factor. In the treatment of bowel perforation by a peritoneal catheter, suspected shunt infection should be managed properly and contamination be minimized during removal of the peritoneal catheter. PMID- 10804057 TI - Ruptured intracranial mycotic aneurysm presenting as cerebral haemorrhage in an infant: case report and review of the literature. AB - A 2-month-old male infant presented with intracranial haemorrhage caused by ruptured intracranial mycotic aneurysm. Computed tomography and cerebral selective angiography revealed a large haematoma in the left sylvian fissure and a mycotic aneurysm of a peripheral branch of the middle cerebral artery. Despite the successful surgical removal, the child did not recover from the initial brain injury and died 2 months later. There have been fewer than 10 reported cases of infantile mycotic aneurysms and its occurrence in the absence of infectious endocarditis is exceptionally rare. PMID- 10804058 TI - Electrical synapses by guided growth of cultured neurons from the snail Lymnaea stagnalis. AB - The ability to assemble neuronal networks with designed topology would allow uniquely defined experiments on neurocomputing. We describe a fundamental step, the controlled formation of synapses by guided outgrowth, in vitro for the first time combining simple neuritic geometry with predefined connectivity. We used neurons from the A-clusters in the pedal ganglia of the snail Lymnaea stagnalis. They were cultured on a substrate with linear patterns made by adsorption of brain-derived conditioning factors and photolithography. We induced and observed the frontal collision of two growth cones on narrow lanes. Following such encounters. individual electrical synapses formed that were sometimes strong enough for prolonged presynaptic stimulation to reach the threshold of postsynaptic firing. PMID- 10804059 TI - Detection of compound spatial patterns: further evidence for different channel interactions. AB - It is shown that contrast interrelation functions for small compound gratings and for small compound edge-type patterns have different shapes: the former are lozenges, the latter ellipses in the normalised contrast space. These findings can be described by a simple p-norm model, comprising one channel which is most sensitive to the first pattern, another channel which is most sensitive to the second pattern and a non-linear summation of the channel outputs. Since the value of the summation exponent p is constant within but varies between pattern classes, an interpretation of the model parameters is limited to pattern class. Channel models based on the p-norm are discussed, taking into account the interpretation of the first derivative of the contrast interrelation function and the summation exponent, p. PMID- 10804060 TI - An interspike interval method for computing phase locking from neural firing. AB - Inner hair cells (IHCs) transform the mechanical movements of the basilar membrane into electrical impulses. The impulse coding of the IHCs is the main information carrier in the auditory process and is the basis for improvements of cochlea implants as well as for low rate, high-quality speech processing and compression. This paper shows how to compute the speech signal from the neural firing based on the analysis of the interspike interval histogram. This new approach solves problems that other standard analysis methods do not solve sufficiently well. PMID- 10804061 TI - A mean-field model for orientation tuning, contrast saturation, and contextual effects in the primary visual cortex. AB - Orientation-selective cells in the primary visual cortex of monkeys and cats are often characterized by an orientation-tuning width that is invariant under stimulus contrast. At the same time their contrast response function saturates or even super-saturates for high values of contrast. When two bar stimuli are presented within their classical receptive field, the neuronal response decreases with the intersection angle. When two stimuli are presented inside and outside the classical receptive field, the response of the cell increases with the intersection angle. Both cats and monkeys show iso-orientation suppression, which has sometimes been reported to be combined with cross-orientation facilitation. This property has previously been described as sensitivity to orientation contrast. We address the emergence of these effects with a model that describes the processing of geniculocortical signals through cortical circuitry. We hypothesize that short intracortical fibers mediate the classical receptive field effects, whereas long-range collaterals evoke contextual effects such as sensitivity to orientation contrast. We model this situation by setting up a mean field description of two neighboring cortical hypercolumns, which can process a nonoverlapping center and a (nonclassical) surround stimulus. Both hypercolumns interact via idealized long-range connections. For an isolated model hypercolumn, we find that either contrast saturation or contrast-invariant orientation tuning emerges, depending on the strength of the lateral excitation. There is no parameter regime, however, where both phenomena emerge simultaneously. In the regime where contrast saturation is found, the model also correctly reproduces suppression due to a second, cross-oriented grid within the classical receptive field. If two model hypercolumns are mutually coupled by long-range connections that are iso-orientation specific, nonclassical surround stimuli show either suppression or facilitation for all surround orientations. Sensitivity to orientation contrast is not observed. This property requires excitatory-to excitatory long-range couplings that are less orientation specific than those targeting inhibitory neurons. PMID- 10804062 TI - Analysis of neural spike trains with interspike interval reconstruction. AB - As a method for the analysis of neural spike trains, we examine fundamental characteristics of interspike interval (ISI) reconstruction theoretically with a leaky-integrator neuron model and experimentally with cricket wind receptor cells. Both the input to the leaky integrator and the stimulus to the wind receptor cells are the time series generated from the Rossler system. By numerical analysis of the leaky integrator, it is shown that, even if ISI reconstruction is possible, sometimes the entire structure of the Rossler attractor may not be reconstructed with ISI reconstruction. For analysis of the in vivo physiological responses of cricket wind receptor cells, we apply ISI reconstruction, nonlinear prediction and the surrogate data method to the experimental data. As a result of the analysis, it is found that there is a significant deterministic structure in the spike trains. By this analysis of physiological data, it is also shown that, even if ISI reconstruction is possible, the entire attractor may not be reconstructed. PMID- 10804063 TI - Alpha activity decreases during the perception of Necker cube reversals: an application of wavelet transform. AB - Since the first observation of perceptual reversal by Necker, many theoretical approaches have been proposed. In a previous study, we showed that a positive wave appeared approximately 250 ms prior to the button press of the subjects, indicating perceptual reversal during the observation of the Necker cube figure. A basic difficulty in this type of study is the possible jitter in the latency of the button press due to the variability of the subjects' reaction time during a recording session. To overcome this difficulty, a pattern selection method based on the wavelet transform was proposed in the previous study. A dominant positive wavelet coefficient in the delta band was found to represent the perceptual reversal-related positivity. In the present study, we aim to analyze the changes in the alpha frequency band during perceptual reversal by using the Necker cube. The RMS values of the alpha frequency band were measured for two time periods: +/ 3 SD around the mean peak latency of the perceptual-reversal-related positivity and a time window of the same length before the positive wave. We found significantly increased delta power and decreased alpha power during the perceptual-reversal-related positivity. PMID- 10804064 TI - Cerebellar learning of accurate predictive control for fast-reaching movements. AB - Long conduction delays in the nervous system prevent the accurate control of movements by feedback control alone. We present a new, biologically plausible cerebellar model to study how fast arm movements can be executed in spite of these delays. To provide a realistic test-bed of the cerebellar neural model, we embed the cerebellar network in a simulated biological motor system comprising a spinal cord model and a six-muscle two-dimensional arm model. We argue that if the trajectory errors are detected at the spinal cord level, memory traces in the cerebellum can solve the temporal mismatch problem between efferent motor commands and delayed error signals. Moreover, learning is made stable by the inclusion of the cerebello-nucleo-olivary loop in the model. It is shown that the cerebellar network implements a nonlinear predictive regulator by learning part of the inverse dynamics of the plant and spinal circuit. After learning, fast accurate reaching movements can be generated. PMID- 10804065 TI - Postural control model interpretation of stabilogram diffusion analysis. AB - Collins and De Luca [Collins JJ. De Luca CJ (1993) Exp Brain Res 95: 308-318] introduced a new method known as stabilogram diffusion analysis that provides a quantitative statistical measure of the apparently random variations of center-of pressure (COP) trajectories recorded during quiet upright stance in humans. This analysis generates a stabilogram diffusion function (SDF) that summarizes the mean square COP displacement as a function of the time interval between COP comparisons. SDFs have a characteristic two-part form that suggests the presence of two different control regimes: a short-term open-loop control behavior and a longer-term closed-loop behavior. This paper demonstrates that a very simple closed-loop control model of upright stance can generate realistic SDFs. The model consists of an inverted pendulum body with torque applied at the ankle joint. This torque includes a random disturbance torque and a control torque. The control torque is a function of the deviation (error signal) between the desired upright body position and the actual body position, and is generated in proportion to the error signal, the derivative of the error signal, and the integral of the error signal [i.e. a proportional, integral and derivative (PID) neural controller]. The control torque is applied with a time delay representing conduction, processing, and muscle activation delays. Variations in the PID parameters and the time delay generate variations in SDFs that mimic real experimental SDFs. This model analysis allows one to interpret experimentally observed changes in SDFs in terms of variations in neural controller and time delay parameters rather than in terms of open-loop versus closed-loop behavior. PMID- 10804066 TI - The effect of intracortical competition on the formation of topographic maps in models of Hebbian learning. AB - Correlation-based learning (CBL) models and self-organizing maps (SOM) are two classes of Hebbian models that have both been proposed to explain the activity driven formation of cortical maps. Both models differ significantly in the way lateral cortical interactions are treated, leading to different predictions for the formation of receptive fields. The linear CBL models predict that receptive field profiles are determined by the average values and the spatial correlations of the second order of the afferent activity patterns, whereas SOM models map stimulus features. Here, we investigate a class of models which are characterized by a variable degree of lateral competition and which have the CBL and SOM models as limit cases. We show that there exists a critical value for intracortical competition below which the model exhibits CBL properties and above which feature mapping sets in. The class of models is then analyzed with respect to the formation of topographic maps between two layers of neurons. For Gaussian input stimuli we find that localized receptive fields and topographic maps emerge above the critical value for intracortical competition, and we calculate this value as a function of the size of the input stimuli and the range of the lateral interaction function. Additionally, we show that the learning rule can be derived via the optimization of a global cost function in a framework of probabilistic output neurons which represent a set of input stimuli by a sparse code. PMID- 10804067 TI - If you were an old woman who lived in a shoe, what would you do? PMID- 10804068 TI - Mental health services in long-term care: still an unmet need. PMID- 10804069 TI - Elder maltreatment. PMID- 10804070 TI - Exposing financial exploitation of impaired elderly persons. AB - There is a dearth of medical publications on financial exploitation of elderly persons, but a significant amount of information on this subject is contained in the literature of other disciplines. Financial abuse accounts for up to one-half of all types of elder abuse in the United States, accounting for over 500,000 victims. Psychological abuse, including deception, intimidation, and threats, always accompanies financial exploitation. Despite the devastating emotional and financial losses incurred, physicians are reluctant to recognize, diagnose, and assist impaired elderly victims of financial exploitation. PMID- 10804072 TI - Late-life atypical major depressive episode: a 358-case study in outpatients. AB - The author compared the prevalence and symptoms of DSM-IV major depressive episode (MDE) with atypical features between older and younger MDE outpatients (N = 358). Atypical MDE was present in 55.0% of MDE patients under age 60 and in 28.1% age 60 and over (P = 0.0000). Bipolar II disorder was present in 56.4% of younger patients, and in 23.9% of late-life patients (P = 0.0000). Late-life atypical MDE patients had less interpersonal-rejection sensitivity. Prevalence of atypical MDE seems lower among late-life MDE outpatients than among younger MDE outpatients, which may be related to the decrease with aging in the prevalence of patients with bipolar II MDE, where there is a higher prevalence of atypical features. PMID- 10804073 TI - Stage-specific prevalence of behavioral symptoms in Alzheimer's disease in a multi-ethnic community sample. AB - The authors extended previous studies of the stage-specific prevalence of behavioral pathology to members of ethnic minority groups. Behavioral symptoms and their relationship to severity of Alzheimer's disease (AD) were examined in 125 heterogeneous minority elderly patients interviewed with a modified CERAD protocol, with behavioral symptoms scored on the caregiver-rated BEHAVE-AD Rating Scale. Behavioral disturbances were extremely common, occurring in 98% of the sample; the most common was activity disturbances (89%), followed by paranoid and delusional ideation (72%), aggressivity (64%), anxieties and phobias (61%), depressive symptoms (50%), sleep disturbances (43%), and hallucinations (34%). As in white patients, overall behavioral symptoms were most frequent among patients with moderate and severe dementia. Preliminary evidence supports the possibility of ethnic differences in behavioral profiles; Blacks showed lower affective, anxiety, and sleep symptoms than Asians and Hispanics, and lower total BEHAVE-AD scores than Hispanics. Inquiry in larger, population-based samples will be needed to determine whether the ethnic differences in behavioral symptoms of AD found here are robust and replicable. PMID- 10804071 TI - Maintenance treatment for recurrent depression in late life. A four-year outcome study. AB - The authors examined the 4-year outcome of elderly patients who were given open label maintenance treatment for recurrent depression. Thirty-eight patients, age 60 years or older, who had recovered from an episode of recurrent nonpsychotic unipolar major depression were maintained on full-dose antidepressant medication and, if necessary, adjunctive lithium. They were followed on a regular basis for 4 years or until recurrence, whichever occurred first. The cumulative probability of remaining well without recurrence was 70%. Longer time to respond to treatment and higher anxiety scores at the time of response predicted shorter time to recurrence. PMID- 10804074 TI - The relationship between donepezil and behavioral disturbances in patients with Alzheimer's disease. AB - The authors tested the hypothesis that behavioral disturbances are reported at significantly lower rates by caregivers of Alzheimer's disease (AD) patients receiving the antidementia drug donepezil, compared with a group of patients receiving no antidementia drug treatment. Patients administered donepezilfor 6 months (n=84) were compared with patients not on donepezil (n=248). Patients taking donepezil had significantly lower levels of behavioral disturbances than patients not receiving this agent (P< or =0.011). Specifically, donepezil patients were described as significantly (P< or =0.05) less likely to be threatening, destroy property, and talk loudly. Also, significantly fewer patients receiving donepezil were treated with sedatives (P< or =0.005). These findings support the growing body of evidence that cholinesterase inhibitors have psychotropic properties and reduce behavioral disturbances in patients with AD. PMID- 10804075 TI - Heuristic comparison of sertraline with nortriptyline for the treatment of depression in frail elderly patients. AB - Studies have demonstrated that the selective serotonin reuptake inhibitor antidepressants have similar efficacy to other agents, such as tricyclic antidepressants. However, data are limited for direct comparisons with other antidepressants. The authors conducted a contemporaneous comparison of nursing home residents treated with open-label sertraline in doses up to 100 mg/day with nursing home residents treated in a double-blind randomized study of low vs. regular doses of nortriptyline. There were 97 patients enrolled in the study (28 treated with sertraline), with an average treatment duration of 55 days. There were no differences in the tolerability of sertraline vs. nortriptyline. However, in this group of frail older adults, sertraline was not as effective as nortriptyline for the treatment of depression. PMID- 10804076 TI - Drug treatment of depression in frail elderly nursing home residents. AB - The authors conducted a randomized, double-blind, 10-week clinical trial of two doses of nortriptyline in eight nursing homes. Sixty-nine patients, average age 79.5 years, were randomized to receive regular doses (60 mg-80 mg/day) vs. low doses (10 mg-13 mg/day) of nortriptyline. Among the more cognitively intact patients, there was a significant quadratic relationship defining a "therapeutic window" for nortriptyline plasma levels and clinical improvement. There were also significant differences in plasma level-response relationships between depressed patients who were cognitively impaired and those who were more cognitively intact. Depression remains a syndrome that responds to specific treatment, even in frail nursing home patients, and those depressions that occur in patients with significant dementia may represent a treatment-relevant condition with a different plasma level-response relationship than in depression alone. PMID- 10804077 TI - Mental health service use by elderly patients with bipolar disorder and unipolar major depression. AB - Symptoms, functioning, and mental health service use were compared in older out patients with bipolar disorder and unipolar depression. Bipolar outpatients (n = 37, mean age = 69.7) had higher total symptom severity and positive symptom scores, more impaired community-living skills, and earlier age at onset of illness than patients with unipolar depression (n = 85, mean age = 70.9). Bipolar elderly patients used almost four times the total amount of mental health services and were four times more likely to have had a psychiatric hospitalization over the previous 6 months. These findings underscore the need for effective services for elderly patients with bipolar disorder, who account for a minority of patients with affective disorders, but use a disproportionate amount of costly services. PMID- 10804078 TI - Primary vs subspecialty care: a structured follow-up of dementia patients and their caregivers. AB - All dementia patients and their caregivers who had received a University-based comprehensive evaluation and a diagnosis of Alzheimer's disease during 1997 (N = 80) were surveyed 1 year after their initial assessment. Of the original cohort, 72.5% were contacted, and two subgroups were defined: 31 patients were being seen only by their primary care physicians (MED), and 27 patients were being treated in addition by a geriatric psychiatry faculty member (GERO). There were statistically significant differences between the two groups (MED vs. GERO, respectively) at follow-up in terms of: 1) hospitalization (39% vs. 15%; P 0.05); the patients living rural area had mode sensitive to weed and tree pollens than those from city center (60.7% versus 45.8% for weeds and 42.9% versus 26.4% for trees, p < 0.05). The results suggested that sensitization to pollens in respiratory system allergies might be related to differences in macro (ie: climate) and micro (city or rural area) environmental characteristics of an area. PMID- 10804094 TI - Specific sublingual immunotherapy in atopic dermatitis. Results of a 6-year follow-up of 35 consecutive patients. AB - BACKGROUND: allergen-specific immunotherapy has proved to be effective in selected patients with IgE-mediated respiratory allergic diseases, and alternative routes of administration are being studied. Atopic Dermatitis (AD) is currently regarded as an allergic inflammatory disease. METHODS: we conducted a cohort study to evaluate the safety and effectiveness of sublingual-swallow immunotherapy (SLIT) in selected patients with allergic (extrinsic) AD. Thirty five patients, 16 suffering from AD without respiratory allergic symptoms (Group A) and 19 with AD associated to mild asthma and/or rhinitis (Group B), were enrolled in the study. The severity of the skin lesions (eczema) was scored on a 0 to 4 scale (and subsequently related to the more recent SCORAD Index), where 0 indicated complete healing of the eczema and 4 indicated maximal spread of the lesions. Only patients with an eczema score of 1 to 3 were started on allergen specific SLIT for 36 months. Eczema scores, symptoms and side effects were recorded every two months during the first 2 years and then after 36 months. After SLIT was completed, all patients attended 3 yearly follow-up visits to evaluate the long-term effects of the treatment. All patients followed a set of rules designed to control for identified confounding variables. All patients received ketotifen during the first 3 months of SLIT. RESULTS: only the complete disappearance of skin lesions (score 0) was considered to indicate effectiveness. In Group A this was observed in 12.6% of the patients after 6 months of SLIT, in 31,2% after 12 months and 68.8% after 24 months. In Group B, eczema disappeared in 0% after 6 months, in 36.8% after 12 months and 73.7% after 24 months. No patients in Group A developed asthma during SLIT, and 1 patient developed asthma 3 years after immunotherapy had ended. Three focal reactions consisting of 2 cases of mild eczema and one case of diarrhoea were recorded. One case of urticaria, due to violation of the administration schedule was the only systemic reaction observed. No life-threatening reactions appeared at any time of the study. CONCLUSIONS: the outcomes obtained, taken into account the limitations of the study design, suggest that sublingual allergen-specific immunotherapy for the treatment of the extrinsic form of Atopic Dermatitis is safe and well tolerated by patients, and may favourably affect the natural course of the disease. PMID- 10804096 TI - What does Anisakis simplex parasitism in gastro-allergic anisakiasis teach us about interpreting specific and total IgE values? AB - BACKGROUND: gastro-allergic Anisakiasis is a mostly transitory clinical entity caused by Anisakis simplex (A. simplex) and can be suspected by history and confirmed by fiberoptic gastroscopy and specific IgE. OBJECTIVE: we report a case of gastro-allergic Anisakiasis, in which the parasite induces a high specific and total IgE response, and want to follow the specific and total IgE values by a serologic follow up over 10 months. METHODS: an analysis of total IgE and specific IgE against. A. simplex was performed within 24 hours, after 1, 4, 6 and 10 months. At month 4 and month 10 specific IgE against Ascaris lumbricoides and Echinococcus granulosus was determined in order to value cross-reactivity. RESULTS: there is an important raise in specific IgE against Anisakis simplex (up to 903 kU/l) after 6 months and total IgE (up to 15,258 kU/l) after one month. Cross-reactive specific IgE against Ascaris lumbricoides and Echinococcus granulosus can be detected. CONCLUSIONS: we consider a raise of total and specific IgE as a typical feature of helminth infestation and learn that specific and total IgE values are highly variable in the months following the allergic and parasite-specific reaction. The amount of specific IgE against other cross reactive parasites depends directly on the total IgE values. PMID- 10804097 TI - Clinical study on Alternaria spores sensitization. AB - BACKGROUND: sensitization to fungi spores is often associated with sensitization to other allergens and so it is difficult to estabilish the exact cause of illness. OBJECTIVE: the aim of the study was to evaluate a group of patients monosensitized to Alternaria and to establish the prevalence, periodicity and clinical profile of this kind of sensitization. METHODS: 37 subjects who were monosensitized to Alternaria (prick-test positive), according to a previous epidemiological study were evaluated. Clinical symptoms and the period of their occurrence were taken into consideration as well as immunological parameters (RAST). RESULTS: 20 patients (over 50%) included in the study proved to be affected by asthma associated with other allergic symptoms and 22 patients (60%) presented perennial symptoms. The RAST carried out on 34 monosensitized subjects proved positive in 11 and negative in 23. CONCLUSIONS: Alternaria sensitization is characterized by a perennial periodicity with severe respiratory symptoms (asthma) which occur primarily in children. Prick test is preferable to and more reliable than RAST as a diagnostic test. PMID- 10804098 TI - Aluminium allergy in a patient with occupational contact dermatitis. AB - Case of a 57-year-old hospital attendant with hand eczema. Patch tests were read at 2 and 3 days using the 1+ to 3+ scoring system recommended by the Contact Dermatitis Research Group. PMID- 10804099 TI - [Time trends in cancer incidence in Navarra and Zaragoza, spain]. AB - BACKGROUND AND OBJECTIVES: The knowledge of time-related changes occurring in cancer incidence and mortality is an essential element for cancer control. This paper aims to describe the time trends of the more important cancer sites in Navarra and Zaragoza. METHODS: The influence of age, diagnosis period and birth cohort on the observed time trend cancer incidence in Navarre and Zaragoza was assessed using a log-linear model. Results are showed graphically, for the different tumour sites by sex and each registry. RESULTS: Among males, a considerable rise in the incidence of lung cancer (5% per year), prostate cancer (> 2% per year) and non-Hodgkin's lymphomas was found. Among females, the highest increases corresponded to non-Hodgkin's lymphomas, with a yearly rise greater than 7%, and ovarian cancer (4% per year). Breast cancer in women increased in both registries (3.5% per year in Navarre and 0.9% per year in Zaragoza), part of the increment in Navarra being explained by a higher case detection rate. Finally, the incidence of colorectal, bladder and kidney cancer rose more than a 3% per year in both registries and sexes. For most types of tumours, cancer risk increased with subsequent generations. CONCLUSION: The substantial cancer increment observed points out the inefficacy of primary prevention policies, the importance of studying cancer incidence for long time periods and the need to increase the population coverage of Spanish cancer registries. PMID- 10804100 TI - [Prevalence of tuberculosis infection in the school age in Guadalajara, Spain]. AB - OBJECTIVES: To estimate the prevalence of tuberculosis infected children in the first grades of Primary Education and to analyse the distribution of infected people among social class groups and other previous circumstances. METHODS: Cross sectional study with diagnostic test application and questionnaire survey, in a representative sample of the school population of the province of Guadalajara, Spain (two stages sampling design). Tuberculin skin tests with 2 I.U. of P.P.D. RT-23 and the Sokal method for lecture were used. For occupational classification and social class definition were used the criteria of the last report of Spanish Epidemiology Society. RESULTS: A total of 1,093 children (50.8% males) between 6 and 9-year-old were studied. The overall prevalence of infected children was 1.64 (I.C.:1-2.3). The distribution of infection did not show significant differences between urban/rural population, previous disease, family antecedent, social class, dwelling area and occupational activities of parents. The prevalence increased in the older upper school years and among the older children. Prevalence ranged from 0.4% (CI: 0.01-1.3) in the first year to 3.53% (CI: 2.4 4.6) in the last year. Children born in 1988 had a 3.23% prevalence (CI: 1.2 5,1), while those born in 1989; 1990 and 1991 had a prevalence of 0.73 (CI: 0 1.77), 1.88% (CI: 0.2-3.51) and 0.4% (0-1.2), respectively. CONCLUSIONS: Tuberculosis infection is less prevalent at school ages than other areas of Spain. In spite of the increase prevalence per school year, the low frequency and sample size did not allow to test for other associations. PMID- 10804101 TI - [Characteristics of patients who do not use primary care services]. AB - OBJECTIVE: To determine the proportion of patients who do not consult their doctor for one year time and to investigate their characteristics, to determine of they are different from patients that do consult. METHODS: This is a prospective study, with a follow-up of one year in the city of Valencia, Spain. The patients included were all registered in a general practitioner's list, 1473 subjects. We took every day all the patients that consulted: home visits were excluded, and so were visits from patients that did not belong to our territory, patients of another general practitioner's list and all patients under 14 years. We included patients attended without citation. We counted the number of visits for each subject, and their age and gender, presence of hypertension, diabetes, hyperlipidemia, chronic respiratory disease, HIV/Aids and health problems that limit their autonomy. RESULTS: 46.3% of patients did not consult, 48.7% made between 1 and 14 visits, and 4.9% 15 visits or more (high utilization is defined as the average plus 2 standard deviations). Among the subjects that did not consult, there were significantly less women, they were younger and had less chronic problems than the patients that consulted and these ones less than high users. CONCLUSIONS: The percent of subjects that did not use primary care medical consultations is 46.3% in one year; they were more frequently men, young people and persons without chronic health problems. PMID- 10804102 TI - [Factors related to inappropriate referral between primary and specialized care: qualitative study among primary care physicians]. AB - OBJECTIVE: To characterise the factors that, from a primary care physician's point of view, are related to inappropriate referral from primary to specialised health care. METHODS: We conducted two focus groups and two semi-structured interviews with primary care physicians. Each of the participant's profiles was defined according to variables related to inappropriate referral found in previous studies. Discussions were recorded on audio tape and later transcribed verbatim onto paper. We analysed the factors related to inappropriate referral according to frequency and capacity for generating discussion. RESULTS: Primary care physicians have different concepts regarding inappropriate referral, besides health problems that can be solved in primary care. Inappropriate referral is usually justified. Factors related to inappropriate referral can be divided into four groups: 1) related to the patient and patient/doctor relationship: pressure exerted on the primary care physician, caused by a belief in specialists' greater competence, the right of the patient to specialist referral and mass media pressure; 2) related to the health system: lack of coordination between care levels, consultancy time pressure, lack of equipment and distance to the specialist; 3) related to primary care physicians: lack of training and defensive medical practise; 4) related to specialists: professional competence and behaviour with patients. CONCLUSIONS: A notable gap can clearly be seen between primary and specialised care, and this generates problems in the health system. It is difficult to implement prevention and control measures with the factors related to inappropriate referral. Primary care physicians form opinions that are not based on available evidence. Further research is needed in both qualitative and quantitative fields. PMID- 10804103 TI - [Prevention of drug abuse in the secondary school after the reform of the Spanish educational system. Implementation of several programs in the city of Barcelona, Spain]. AB - OBJECTIVES: To evaluate the adoption of 3 different school-based drug abuse prevention programmes in secondary schools, analyzing some features of their implementation and the relationship between the percentage of schools adopting each intervention and the prevalence of addiction to opiates at district level. METHODS: Mail survey with telephone follow-up to the 235 secondary schools located in Barcelona, carried out at the end of 1997-1998 school-year. A bivariate analysis of correlations (Spearman) was performed to assess the association between the percentage of schools having adopted each of the 3 prevention programmes in each city district, and the previously reported estimates of the prevalence rates of opiate addiction. RESULTS: 44.7% of all schools completed the survey, 38 of them (16.2% of respondents) had offered 1 of the programmes within the school-year, 20 schools (19.0%) offered 2 programmes, and 4 schools (3.0%) the 3 programmes. Statistically significant correlations were found for the adoption of different programs between districts, but not for the percentages of schools offering prevention programmes and the prevalence of opiate addiction. CONCLUSIONS: Social and health needs, indicated by prevalence rates of opiate addiction, do not explain geographic inequalities in the adoption of prevention programmes. The need for prevention programmes aimed at risk groups is discussed. PMID- 10804104 TI - [News about cancer in the press]. AB - BACKGROUND: News on health and health care are increasingly common in the mass media, and cancer is one of the most prominent topic among these news due to its health impact. MATERIAL AND METHODS: All news published in major newspapers in Catalonia, Spain between April to December 1996 were analysed, together with items from science and health supplements general information and science journals. Types of cancer-related topics, sources of information, the importance of cancer in the news item and the theme of the piece were analysed. RESULTS: During the period analysed, 974 news items were published which dealt with cancer. Of these, cancer was the principal topic in 53.5% of the items published in the national press, and 63.4% of the items published in the regional press. Cancer was the subject of an editorial comment on only four occasions. The source of information was mentioned in 64.8% of the items, and mean number of sources was 1.6. Political institutions were the most frequent source of information in newspapers, while science and health supplements and other magazines tended to cite scientific and health care institutions, as well as scientific journals. Basic research was the most common topic for news (30.3%) in science and health supplements while risk and environmental factors (29.6%) were the most common topics in magazines; environmental and technological risk factors (18,6%) and conflicts with health administrative bodies (16%) were the most frequent topics in the newspapers. CONCLUSION: News about risk factors and their prevention as well as technological and environmental factors related to cancer were among the most important topics published in the general press and magazines, together with conflicts with administrative bodies. Informations related to new therapies were a less frequent topic. Scientific journals have an important role as an information source for mass media together with political and scientific institutions. PMID- 10804105 TI - [The importance of the gender perspective in health interview surveys]. AB - The identification and measurement of the population health needs should be the first step in health planning. In order to guarantee equity criteria, to know the situation of the whole population, and therefore also that of women, is a key issue. Health interview surveys are a good tool for pinpointing the needs of the population, but mainly they are usually focused on health risk factors that explain men's health status such as health behaviours and paid job. These factors often fail to capture aspects that are relevant for women's health, such as household work. The main objective of this paper is to emphasise the importance of a gender perspective in the design and analysis of health interview surveys, and to propose variables that should be included in health surveys in order to better know gender health inequalities. Likewise, this article deals with the gender concept and its importance as a health inequality factor. Gender is an analytical construct based on the social organisation of the sexes that can be used to better understand the conditions and factors influencing women's and men's health beginning by the social roles that each culture and society assigns to people based on their sex. Health is a complex process determined by a wide range of factors: biological, social, environmental and health services related factors. Gender, because of its close relation to all of them, plays a key role. The gender approach is characterised by the analysis of the social relation between men and women, taking into account that sex is a determinant of social inequalities. This paper presents the variables that health interview surveys should include from a gender approach point of view: reproductive work, productive work, social class, social support, self-perceived health status, quality of life, mental health and chronic conditions. In addition, issues related to the wording of questions, data collection and analysis are discussed. PMID- 10804106 TI - [New regimen for protection of health data]. AB - The new Personal Data Protection Act of 1999 (Ley Organica 15/1999, de 13 de diciembre) introduces changes in legal protection of health data. This regulation fulfills the Directive 95/46/CE. There are four relevant changes: 1) extension of public control over any processing of data, automatic or manual, except in personal or domestic activities; 2) general prohibition against processing data, unless the subject gives his or her explicit consent, waived when processing personal data is required for medical needs or the management of health care services by any health professional subject to professional secrecy; 3) regardless of the purpose for which the data was been collected, further processing for scientific purposes is considered compatible; 4) any regulations contained in the General Health Act, or any other Act, relating to data concerning health continue in force. In short, the new Act waives previous and explicit consent for processing health data when medical, public heath and research goals are involved. PMID- 10804107 TI - [The measurement of health related-quality of life in clinical research and practice]. PMID- 10804108 TI - [Quality of life and allocation of health care resources]. PMID- 10804109 TI - [Determination of plasma ciprofloxacin levels in children treated with 0.2% topical ciprofloxacin for tympanic perforation]. AB - Topical drug use can produce locally adverse effects by direct action or systemic effects as a result of drug absorption. Local tolerance of topically-administered ciprofloxacin (ear drops) and serum antibiotic levels after 7-10 days of treatment were evaluated in 30 children with tympanic membrane perforation and suppuration. There were no signs of local intolerance or ototoxicity and significant serum ciprofloxacin levels were not detected. We conclude that ciprofloxacin ear drops can be safely used in children with suppurative otitis media and tympanic perforation. PMID- 10804110 TI - [Myringoplasty in chronic simple otitis media. Comparative analysis of underlay and overlay techniques]. AB - Myringoplasty is a surgical procedure that has been routinely performed by otological surgeons for more than 40 years. Underlay and overlay are the most commonly used tympanoplastic techniques. A comparison was made of anatomic and functional results, and the year of intervention, for 97 myringoplasties, 54 using the underlay technique and 43 using the overlay technique. Selection of the overlay or underlay technique was guided by specific indications. Results were evaluated at least 1 year after surgery. No significant differences were found in anatomic success (88.8% for underlay and 97.6% for overlay). Favorable functional results (average gap of 0 to 20 dB for frequencies of 500, 1000, 2000, and 4000 Hz) were obtained in 88.4% of patients treated with the overlay technique and in 83% of those treated with the overlay technique, with no significant difference. We believe that the high percentage of good results was due to careful preoperative selection. PMID- 10804111 TI - [Prospective follow-up of 27 tumors of the oropharynx]. AB - The diagnosis of an oropharyngeal tumor should be simple because the site is easily accessible. However, these tumors are not often diagnosed is early stages, even when lesions are visible. We followed-up 27 oropharyngeal tumors from 1 January 1996 to 31 December 1997. Etiological data, the characteristics of the problem leading to diagnosis, and time from symptoms to definitive diagnosis were studied. Histopathological differentiation and stage determination were analyzed. Therapeutic strategy and post-treatment evolution were studied, among other parameters. Our results suggest that the most important etiopathogenic factors in the development of oropharyngeal carcinoma were alcohol and tobacco use, to effects of which are potentiated at this site. The most frequent primary oropharyngeal site was the tonsils in 58% of patients. Treatment of each tumor should be individualized. The general prognosis is poor because 40% of patients are stage IV. PMID- 10804112 TI - [Orbital cellulitis in childhood. Medical-surgical treatment]. AB - Orbital cellulitis is an uncommon complication resulting from a spectrum of disorders commonly found in pediatric practice. It usually occurs as a complication of infection of the paranasal sinuses, although it also can be caused by eyelid or dental juries, dental infection and external ocular infection. We studied the clinical, microbiological, and therapeutic features of 152 children diagnosed as periorbital cellulitis and 27 children with orbital cellulitis admitted to our hospital in a 16-year period from January 1983 to December 1998. Twenty-four percent of patients (43 cases) had positive cultures. Thirty children with septal or preseptal cellulitis developed neurological or ophthalmological complications. Intravenous or oral antibiotic administration was effective in 150 patients, but a significant proportion required surgery of the paranasal sinus or orbit (16%). PMID- 10804113 TI - [Prognostic factors in supraglottic laryngeal cancer: a review of 74 cases]. AB - Laryngeal carcinoma is a frequent neoplasm in Spain, constituting about 5.6% of all malignant tumors. The supraglottic location is more frequent than either glottic or subglottic sites. Prognostic factors were analyzed in a series of 74 supraglottic laryngeal carcinomas. Two statistical methods were used. Firstly, Cox regression, a multivariate analysis that defines a survival function of the patients, was used to detect variables with prognostic influence. Secondly, mortality tables were studied for all of them. This univariate method is used to determine the number of patients for every option of a study variable and their survival. The most important prognostic variables in our series were: tracheostomy before surgery, pathological study of the cervical nodes, time from surgery to radiotherapy, type of biopsy, and alcohol use. PMID- 10804114 TI - [Study and incidence of carcinoma of the larynx in women]. AB - Recent publications show an increase in the frequency of laryngeal carcinoma in women. Previous studies of patients in our health district (1875-1994) showed a male/female ratio of 56/1. However, a retrospective study of all laryngeal carcinomas diagnosed in a recent year (June 1997-May 1998) showed an overall male/female ratio of 10/1. Women are older at time of diagnosis than men (mean 70.33 years). None of the women was a regular drinker and only one smoked. Glottic location and absence of metastatic lymph node enlargement predominated at time of diagnosis. PMID- 10804115 TI - [Immunohistochemical expression of P53 suppressor genes and retinoblastoma in laryngeal carcinoma]. AB - Carcinoma of the larynx is responsible for 1% of all tumors and for 25% of oncologic disease of the head and neck. Tumor suppressor genes control cell proliferation by suppressing specific mechanisms that stimulate the proliferative response. Rb and p53, both nuclear, are the most studied tumor suppressor genes. We evaluated the immunoexpression of these markers in 72 carcinomas of the larynx and correlated it with patients' clinical parameters: age, sex, occupation, tobacco and alcohol use, marital status, etc. Biological and anatomopathological parameters (tumor stage, histological type, degree of local invasion, and lymph node metastases) also were studied. Statistically significant differences were found in mean immunostaining for the Rb gene with respect to histological grade, lymph node enlargement, and smoking. We observed no positive correlation between p53 and the classic clinical-pathological parameters, although important trends were evident, as well as relatively frequent immunostaining (about 40%) of the total neoplastic population. PMID- 10804116 TI - [Reproducibility of the qualitative interpretation of dynamic salivary radionuclide scans with excretory stimulation]. AB - OBJECTIVE: To evaluate intraobserver and interobserver agreement in the qualitative interpretation of dynamic salivary radionuclide scans to detect salivary disease, and the agreement between interpretation of dynamic and static image compositions. MATERIAL AND METHODS: Two observers (A, B) interpreted 110 dynamic salivary radionuclide scans with excretory stimulation based on dynamic image composition. Both again interpreted every studies: A based on dynamic image composition and B on static image composition. Kappa statistics were used to determine the degree of intraobserver and interobserver agreement. RESULTS: Considering all the studies, the dynamic composition showed an intraobserver agreement of 0.76 and interobserver agreements of 0.58 and 0.61. In patients with clinical sicca syndrome, agreements were 0.60, 0.52 and 0.62, respectively. For all the cases, the agreements between dynamic and static composition was 0.62 (intraobserver), and 0.42 and 0.43 (interobserver). For patients with clinical sicca syndrome these agreements were 0.63 (intraobserver), and 0.36 and 0.51 (interobserver). CONCLUSION: Intraobserver and interobserver agreement in the qualitative interpretation of dynamic salivary radionuclide scan with excretory stimulation are moderate and notable-moderate overall and in patients with clinical sicca syndrome. These agreements are greater than between dynamic and static image composition. PMID- 10804117 TI - [Second primary tumors in patients with carcinoma of the head and neck]. AB - With improved loco-regional control of cancer of the head and neck, second primary tumors (SPT) have an increasingly negative impact on survival. The appearance of SPT depends on the location and extension of the primary and is associated with lower survival. SPT were studied in a series of 1493 previously untreated patients with carcinoma of the head and neck and a minimum follow-up of 2 years. SPT occurred in 6.3% (96/1493). SPT were associated with the following primary tumor sites: 10.8% (57/727) larynx, 4% (6/153) hypopharynx, 4% (16/404) oral cavity, and 4% (6/153) oropharynx. Forty-three percent (42.7%) of SPT were carcinomas of the head and neck, 27% bronchogenic carcinomas, and 17.7% esophageal carcinomas. Survival at 6 months, 1 year, 3 years, and 5 years of patients diagnosed as SPT was 62%, 43%, 21%, and 16%, respectively. PMID- 10804118 TI - [Multiple primaries of the upper aerodigestive tract, esophagus, and lung]. AB - The existence of multiple primary tumors of the upper aerodigestive tract, esophagus, and lung (UADT-E-L) is related with a common etiopathogenic factor (alcohol-tobacco association). In a review of 43 patients, 6.75% with a UADT-E-L tumor developed another neoplasm, 3.25% at the same site. Nine percent (8.59%) of the tumors were synchronic and 10.85% were metachronic. The most frequent association was larynx-lung. Another neoplasm was detected by physical examination and/or radiology in 44.18% of cases, with a time lapse of less than 3 years in 50%. The most frequent treatment was surgery with/without complementary radiotherapy. The most common stage was T1-T2 (62.06%) and N0-N1 (68.96%). The survival rate was 31% at 2 years and 25% at 3 years. PMID- 10804119 TI - [Otogenic cerebral abscess: a persistent problem]. AB - A young man with otitis media and cholesteatoma of the left ear developed secondary cerebral abscess. The clinical debut was non-specific, with headache, mild fever, and mild persistent otalgia in spite of early antibiotic treatment. Studies revealed a cerebral abscess, so ENT surgery in collaboration with the neurosurgery department was decided. This case illustrates that clinical manifestations in such cases can be mild and highlights the need to exclude this type of serious pathology. PMID- 10804120 TI - [Inflammatory pseudotumor of the maxillary sinus: report of a new case and review of the literature]. AB - A new case of inflammatory pseudotumor (IPT) of the maxillary sinus is presented. The clinical and histological features of this rare entity are discussed. Immunohistochemical studies support a possible myofibroblastic origin. Early diagnosis is important to prevent IPT, classified as a benign disease, from becoming a progressive destructive lesion. PMID- 10804121 TI - [Pneumoencephalus of late onset secondary to frontal sinus fracture]. AB - We report a case of late-onset pneumoencephalus after fracture of the frontal sinus. Long-term follow-up sinus injuries is recommended. PMID- 10804122 TI - [Nasal leishmaniasis in an HIV-positive patient]. AB - Leishmania is currently one of the most clinically important protozoa in otorhinolaryngology. Mediterranean countries, including Spain, have endemic HIV and L. infantum. Dogs are the most important Leishmania reservoir. Leishmaniasis is transmitted basically by the bite of infected female Phlebotomus sandflies. Its clinical development depends mainly on the host's cellular immunity (TCD4+ lymphocyte count). About 400 cases of HIV-visceral leishmaniasis have been reported in Spain. However, exclusively cutaneous presentation of HIV leishmaniasis coinfection has been observed in only 2-3% of cases. We report the case of a female HIV+ patient who developed cutaneous leishmaniasis of the nasal vestibule by L. infantum. The patient was treated satisfactorily with a combination of parenteral Pentostam (sodium stilbogluconate) and periodic intralesional injections of Pentostam. The patient was included in a secondary prophylaxis protocol for visceral leishmaniasis with a monthly dose of Glucantime (meglumine antimoniate) for life. PMID- 10804123 TI - [Laryngeal herpes. A case report]. AB - An interesting case of laryngeal herpetic involvement simulating a neoplasm in its gross appearance and the production of paralysis is reported. The patient was diagnosed as thymoma three months later. The possibilities of differential diagnosis and relation with immunosuppression states are discussed. PMID- 10804124 TI - [Small-cell carcinoma of the larynx]. AB - Primary small-cell (also known as oat-cell) neuroendocrine carcinoma of the larynx is an infrequent tumor. We report a case of epiglottic oat-cell carcinoma in a 57-year-old man. Its association with other malignant neoplasms and aggressive behavior in spite of treatment are discussed. PMID- 10804125 TI - [Diffuse deep cervical infection. Report of three cases]. AB - Deep cervical infections still appear despite antibiotic use. Necrotizing fasciitis of the head and neck is a rare but serious infection caused by mixed, predominantly anaerobic bacterial flora. The natural course of the disease is spreading necrosis of the soft tissues of the neck and septicemia. Three recent cases of cervical necrotizing soft-tissue infection are presented. The importance of early and aggressive surgical treatment complemented by parenteral antibiotics with an aerobic-anaerobic spectrum is emphasized. PMID- 10804126 TI - [A prolonged case of intractable sneezing. Pathophysiological hypothesis]. AB - A 13-year-old boy sneezed paroxysmally and continuously, day and night, for 18 days. Sneezing limited his daily activities and motivated hospital admission. Pediatric clinical examination and all other immunologic, otorhinolaryngologic, pneumologic, neurologic, radiologic, and psychiatric studies were normal. There was no response to treatment with antihistamines, corticoids, or anti convulsants, which obscured the diagnosis. Definitively effective treatment and its implications in the pathophysiology of sneezing are discussed. PMID- 10804132 TI - Introduction: human enteric caliciviruses-an emerging pathogen whose time has come. PMID- 10804133 TI - Summary of the first international workshop on human caliciviruses. PMID- 10804134 TI - The epidemiology of enteric caliciviruses from humans: a reassessment using new diagnostics. AB - In the United States, acute gastroenteritis is one of the most commonly noted illnesses on hospital discharge records and death certificates, yet few of these cases have an etiologic diagnosis. The application of new molecular diagnostic methods has shown caliciviruses (previously referred to as the Norwalk family of viruses or small round structured viruses) to be the most common cause of acute gastroenteritis (AGE) outbreaks in the United States, and they may emerge as a common cause of sporadic cases of AGE among both children and adults. Novel molecular methods have permitted outbreak strains to be traced back to their common source and have led to the first identification of virus in implicated vehicles of infection-water, shellfish, and foods contaminated both at their source and by food handlers. The broad application of these methods to routine diagnosis in hospitals and public health laboratories is advancing our appreciation of the full burden of calicivirus-associated diarrhea, and it is opening new avenues for its prevention and control. PMID- 10804135 TI - Molecular epidemiology of human enteric caliciviruses in The Netherlands. AB - In The Netherlands, illness due to Norwalk-like viruses (NLVs) of the family Caliciviridae is quite common. NLVs cause >80% of the outbreaks of gastroenteritis reported to municipal public health centers and at least 5% of the cases of gastroenteritis for which a general practitioner is consulted. In addition, up to 18% of community cases of gastroenteritis in the 1998/1999 winter season have been attributed to NLVs. Patterns of disease activity differ remarkably, with "normal years, when outbreaks occur that are caused by different types of NLV, and "epidemic years, when outbreaks appear to be caused by a single strain. This observation suggests selection of antigenic variants with increased virulence or altered modes of transmission. In addition, since caliciviruses related to the NLVs from humans have been detected in stool specimens from calves at 45% of the dairy farms in The Netherlands, the possibility of spillover of epidemic strains from an animal reservoir to humans should be considered. PMID- 10804136 TI - Surveillance of viral gastroenteritis in Japan: pediatric cases and outbreak incidents. AB - Surveillance results from pediatric cases and outbreaks of viral gastroenteritis in Japan are presented. In winter, both small round structured virus (SRSV, or Norwalk-like viruses) and rotavirus were detected from infants with gastroenteritis; however, in recent years, the prevailing time of SRSV infection has preceded that of rotavirus infection. Most nonbacterial gastroenteritis outbreaks were related to SRSV infection, and >60% of the outbreaks were caused by contaminated food. In small-sized outbreaks, raw oysters were the primary source of transmission. In large-sized outbreaks, school lunches and catered meals that were served at schools, banquet halls, and hospitals were most often implicated in the transmission of foodborne gastroenteritis. PMID- 10804137 TI - Epidemiology of calicivirus infections in Sweden, 1994-1998. AB - Outbreaks of acute gastroenteritis are frequently caused by caliciviruses. Electron microscopy was used to search for these viruses in fecal samples from patients with acute gastroenteritis. Of 5800 samples collected and analyzed from November 1994 to June 1998, 3700 were associated with outbreaks. A total of 676 outbreaks were analyzed, and viruses were found in 67%. Caliciviruses, usually Norwalk-like viruses (NLVs), were found in 407 (89%) of 455 outbreaks, while Sapporo-like viruses were identified in nine outbreaks, including six that were suspected to include foodborne transmission. Sixty percent of the 1041 patients with calicivirus infections were between 70 and 90 years of age. Food- and waterborne infections were associated with 66 calicivirus outbreaks. Virus positive outbreaks were documented mainly during winter and spring. The longitudinal survey showed that caliciviruses, and especially the NLVs, cause most nosocomial and community-associated outbreaks in Sweden. PMID- 10804138 TI - The impact of foodborne calicivirus disease: the Minnesota experience. AB - The first outbreaks of Norwalk virus gastroenteritis in Minnesota were confirmed in 1982. Since then, Norwalk-like caliciviruses have been recognized to be the most common cause of foodborne disease outbreaks, accounting for 41% of all confirmed foodborne outbreaks in Minnesota from 1981-1998. Although laboratory confirmation of caliciviruses in stool samples was not attempted in most of these outbreaks, all conformed to epidemiologic criteria for defining outbreaks of Norwalk virus. Since 1996, the availability of polymerase chain reaction testing at the Minnesota Department of Health has allowed for the confirmation of calicivirus infection among patients involved in epidemiologically defined outbreaks of viral gastroenteritis. Results have confirmed the usefulness of characterizing foodborne disease outbreaks by epidemiologic criteria and also confirmed the importance of human caliciviruses as the leading cause of foodborne disease outbreaks in Minnesota. PMID- 10804139 TI - Cold weather seasonality of gastroenteritis associated with Norwalk-like viruses. AB - Norwalk-like viruses (NLVs) are the most common cause of acute nonbacterial gastroenteritis in adults, but little is known about their seasonality. The lack of specific diagnostic tools impeded study of these viruses in the past, and surveys using electron microscopy often grouped NLVs with other unrelated viruses. A search of the scientific literature found eight surveys of gastroenteritis, which were conducted for at least 1 year, that specifically identified NLVs. Unpublished data from laboratories of 4 NLV researchers were also used. These surveys, which were conducted in eight countries, reported sporadic cases and outbreaks of NLV-associated gastroenteritis among all age groups. The monthly occurrence of these cases and outbreaks was plotted, and while transmission occurred year-round in most surveys, a cold weather peak was demonstrated in 11 of the 12 studies. This key epidemiologic feature of the viruses has important implications concerning their mode of transmission and for understanding the etiology of acute gastroenteritis in adults. PMID- 10804140 TI - Human caliciviruses in acute gastroenteritis of young children in the community. AB - Episodes of acute gastroenteritis in prospectively followed children between 2 months and 2 years of age were examined for rotaviruses, enteric adenoviruses, astroviruses, and human caliciviruses, including both Norwalk-like viruses (NLVs) and Sapporo-like viruses (SLVs), using PCR and reverse transcription (RT)-PCR assays. A virus was identified in 60% (502/832) of all episodes and in 85% of the moderately severe or severe episodes. Human caliciviruses were as common as rotaviruses, both being detected in 29% of the cases. NLVs accounted for a 20% etiologic share of all cases; the clinical picture was a moderately severe disease with vomiting as a predominant symptom. SLVs were detected in 9% of the cases, the clinical picture being a mild diarrheal disease. Astroviruses were found in 10% and enteric adenoviruses in 6% of the cases. Diagnosis with PCR and RT-PCR methods increases the detection of all gastroenteritis viruses, particularly human caliciviruses. As a group, human caliciviruses are common causative agents of gastroenteritis in children <2 years of age in Finland, and, of these, NLVs cause more severe disease than SLVs. PMID- 10804141 TI - The discovery of the 27-nm Norwalk virus: an historic perspective. AB - In 1972, a 27-nm virus-like particle was discovered by use of immune electron microscopy (IEM) in an infectious stool filtrate derived from an outbreak of gastroenteritis in an elementary school in Norwalk, Ohio. IEM enabled the direct visualization of antigen-antibody interaction, as the particles were aggregated and coated by specific antibodies. This allowed the recognition and identification of a 27-nm virus-like particle that did not have a distinctive morphology, was low-titered, and was among the smallest viruses known. Serum antibody responses to the 27-nm particle were demonstrated in key individuals infected under natural or experimental conditions; this and other evidence suggested that this virus-like particle was the etiologic agent of the Norwalk gastroenteritis outbreak. The fastidious 27-nm Norwalk virus is now considered to be the prototype strain of a group of noncultivatable viruses that are important etiologic agents of epidemic gastroenteritis in adults and older children. PMID- 10804142 TI - Sapporo virus: history and recent findings. AB - Morphologically distinct caliciviruses of human origin were first found in stools of children with gastroenteritis in 1976. Sapporo virus, or human calicivirus Sapporo, with typical surface morphology was first detected during a gastroenteritis outbreak in a home for infants in Sapporo, Japan, in 1977. Since then, morphologically and antigenically identical virus has been detected frequently in the same institution in association with outbreaks of gastroenteritis. Sapporo virus is widely distributed worldwide, as evidenced by the appearance of antigenically or genetically similar viruses and seroepidemiologic findings. Sapporo virus plays an important role in outbreaks of infantile gastroenteritis and is less important in foodborne outbreaks. Sapporo virus has been approved as the type species of the genus "Sapporo-like viruses in the family Caliciviridae. The history of and recent findings, as obtained by newly developed techniques, about Sapporo viruses are presented. PMID- 10804143 TI - Organization and expression of calicivirus genes. AB - The application of molecular techniques to the characterization of caliciviruses has resulted in an extensive database of sequence information. This information has led to the identification of 4 distinct genera. The human enteric caliciviruses have been assigned to 2 of these genera. This division is reflected not only in sequence diversity but in a fundamental difference in genome organization. Complete genome sequences are now available for 5 enteric caliciviruses and demonstrate that human and animal enteric caliciviruses are phylogenetically closely related. Currently, there is no cell culture system for the human viruses; therefore, studies have relied on heterologous expression and in vitro systems. These studies have shown that in both human and animal viruses the viral nonstructural proteins are produced from a polyprotein precursor that is cleaved by a single viral protease. The purpose of this article is to provide an overview of the current knowledge of genome structure and gene expression in the enteric caliciviruses. PMID- 10804144 TI - Structural studies of recombinant Norwalk capsids. AB - Norwalk virus is the major cause of epidemic viral gastroenteritis in humans. Attempts to grow this human virus in laboratory cell lines have been unsuccessful; however, the Norwalk virus capsid protein, when expressed in insect cells infected with a recombinant baculovirus, spontaneously assembles into virus like particles. The x-ray crystallographic structure of these recombinant Norwalk particles has been determined to 3.4 A, using a 22-A electron cryomicroscopy structure as a phasing model. The recombinant capsids, 380 A in diameter, exhibit a T=3 icosahedral symmetry. The capsid is formed by 90 dimers of the capsid protein, each of which forms an arch-like capsomere. The capsid protein has two distinct domains-a shell (S) and a protruding (P) domain-that are connected by a flexible hinge. Although the S domain has a classical beta-sandwich fold, the structure of the P domain is unlike any other viral protein. One of the subdomains in the P domain formed by the most variable part of the sequence is located at the exterior of the capsid. PMID- 10804145 TI - Taxonomy of the caliciviruses. AB - The International Committee on Taxonomy of Viruses (ICTV) has recently approved several proposals submitted by the present Caliciviridae Study Group. These proposals include the division of the family into 4 new genera designated Lagovirus, Vesivirus, "Norwalk-like viruses (NLVs), and "Sapporo-like viruses (SLVs); the latter 2 genera were assigned temporary names until acceptable names can be determined by the scientific community. The genera have been further divided into the following species: Feline calicivirus and Vesicular exanthema of swine virus (genus Vesivirus), Rabbit hemorrhagic disease virus and European brown hare syndrome virus (genus Lagovirus), Norwalk virus (genus NLV), and Sapporo virus (genus SLV). In addition, the ICTV approved a proposal to remove the hepatitis E virus from the Caliciviridae into an "unassigned classification status. PMID- 10804146 TI - Immunity to calicivirus infection. AB - The evolution of our understanding of immunity to calicivirus infection, using Norwalk virus as the prototype, is discussed in three stages: (1) "ancient times (1972-1978), when human volunteer studies prevailed, (2) the "middle ages (1978 1990), which were characterized by the development and implementation of solid phase immunoassays based on native viral antigens, and (3) "modern times (1990 to present), which began with the cloning of the genome of the noncultivatable 8FIIa strain of Norwalk virus and resulted in a readily available source of recombinant virus-like particles that have revolutionized the study of caliciviruses. Throughout these stages, it has been shown repeatedly that short-term immunity develops to homologous virus. However, the search for determinants of long-term immunity continues. These studies will likely be facilitated by the newest reagents-the noninfectious recombinant virus-like particles-used in the setting of human volunteer studies and large epidemiologic studies. PMID- 10804147 TI - Genetic classification of "Norwalk-like viruses.. AB - Reverse transcription-polymerase chain reaction has been used worldwide for the diagnosis of Norwalk-like virus (NLV) infection, yet a commonly accepted genetic classification scheme has not been established. Amino acid sequences from four regions of open-reading frame 2 (ORF2) were used to analyze 101 NLV strains, including 2 bovine strains. On the basis of this analysis, a genetic classification scheme is proposed that differentiates 99 human strains into 2 major genetic groups consisting of 5 and 10 genetic clusters, respectively. The 2 bovine strains constitute a newly defined third major genetic group composed of 2 putative clusters represented by each strain. This classification scheme is well supported by the analysis of the entire ORF2 sequences from 38 strains selected to represent the genetic diversity of the human strains used above. This scheme should provide a firm scientific basis for the unified classification of NLV strains detected around the world. PMID- 10804148 TI - Diagnosis of human caliciviruses by use of enzyme immunoassays. AB - The application of molecular technologies, such as the expression of viral proteins in baculovirus, has provided a powerful approach to the diagnosis of human calicivirus (HuCV) infections. The baculovirus-expressed HuCV capsid protein self-assembles into virus-like particles, providing excellent reagents for immunologic assays, such as enzyme immunoassays (EIAs). Following the expression of the capsid protein of Norwalk virus, the capsid proteins of 8 other HuCV strains have been expressed in baculovirus. The unlimited supply of baculovirus-produced reagents for HuCVs allows these EIAs to be applied in large scale clinical and epidemiological studies. Both the antigen and antibody detection EIAs are highly sensitive. The antigen-detection EIAs are highly specific, but the antibody-detection EIAs are more broadly reactive. This article reviews baculovirus expression techniques used to produce HuCV capsid antigens, development of EIAs using these antigens, and application of these EIAs in studies of HuCV infection and illness. PMID- 10804149 TI - Detection of norwalk-like virus in shellfish implicated in illness. AB - In the 1990s, Norwalk-like viruses (NLVs) were identified in patient specimens as the primary pathogen associated with shellfish-borne gastroenteritis in the United States. Identification of these viruses from implicated shellfish has been difficult due to inefficient recovery of viruses, natural polymerase chain reaction (PCR) inhibitors in shellfish, and low virus contamination. Recent improvements to the method of detecting NLVs in shellfish include enhanced processing of virus and shellfish samples, application of nested PCR and nucleotide sequencing, and increased knowledge of NLV genetic diversity. Using a newly developed and sensitive method, an NLV G2 strain was identified in 2 oyster samples implicated in a 1998 California outbreak involving 171 cases. NLV capsid primers demonstrated a greater specificity of PCR detection than did polymerase primers. The 175-base viral capsid nucleotide sequences derived from oysters were 100% identical to those derived from a patient stool sample. This finding supports the epidemiologic associations indicating that contaminated shellfish serve as the vehicle for NLV transmission. PMID- 10804150 TI - Norwalk virus vaccines: challenges and progress. AB - Human caliciviruses (HuCVs) are the major cause of outbreaks of acute nonbacterial gastroenteritis throughout the world. An increasing recognition of the clinical significance of these viruses as human pathogens causing foodborne and waterborne disease indicates that an effective vaccine would be useful. This article reviews the current challenges that exist for the development of a vaccine for the HuCVs as well as the status of development of a candidate vaccine. HuCVs are viruses that exhibit a restricted tropism for infection of the gastrointestinal tract of humans, and a volunteer model of infection and disease is available. As pathogens with a restricted host range, the HuCVs are excellent models for understanding the mechanisms that mediate and regulate viral infection of the gastrointestinal tract and mucosal immunity in humans. PMID- 10804151 TI - Public health concerns about caliciviruses as waterborne contaminants. AB - Caliciviruses are disseminated by the fecal-oral route and are found in contaminated surface and ground waters. The US Environmental Protection Agency (EPA) is interested in preventing calicivirus contamination in treated waters used for consumption, and these viruses are on the EPA's "contaminant candidate list" for regulatory consideration in drinking waters. These viruses also present a health threat for recreation and shellfish-growing waters. However, before EPA can make regulatory decisions regarding caliciviruses, significant information and technology needs must be established, including analytical methods for sampling, identifying, and quantifying the viruses; applicability of surrogates to determine their presence; efficacy of water and wastewater treatment or disinfection; waterborne occurrence levels and distribution; dose response; and the viruses' effect(s) on health. Future drinking-water regulations may need to ensure that treatments are adequate to remove caliciviruses from source waters. For recreation and shellfish-growing waters, surrogate indicators and health criteria may need to be based upon establishing risks of exposure to caliciviruses. PMID- 10804152 TI - Multi-state outbreaks of acute gastroenteritis traced to fecal-contaminated oysters harvested in Louisiana. AB - Norwalk-like viruses (NLVs), or small round structured viruses, are known to cause acute gastroenteritis associated with eating contaminated shellfish. Between 1993 and 1996, three oyster-related gastroenteritis outbreaks attributed to NLV occurred in Louisiana. Intensive trace-back and environmental investigations revealed that the overboard disposal of sewage by oyster harvesters into oyster-bed waters was the most likely source of contamination in at least two of the outbreaks. The small infectious dose of NLV, the large quantity of virus particles in stool, and the ability of oysters to concentrate virus particles suggest that oyster-related outbreaks will continue unless strong control measures are established. Efforts to halt improper sewage disposal in oyster-harvesting waters, including overboard sewage discharge, must be undertaken if future outbreaks are to be prevented. PMID- 10804153 TI - Norwalk-like virus infection in military forces: epidemic potential, sporadic disease, and the future direction of prevention and control efforts. AB - The impact of Norwalk-like virus (NLV) infection on military forces is evaluated in this report. NLVs were a major cause of both outbreaks and sporadic disease among crowded US ground troops in the 1991 war with Iraq. NLVs also have been found to be a cause of acute gastroenteritis in other ground and shipboard deployments. Four large outbreaks of acute gastroenteritis were investigated aboard US Navy aircraft carriers between 1992 and 1997. In these outbreaks, NLVs were identified as the probable cause, and crowding was a major risk factor for transmission. An evaluation of a routine shipboard deployment also suggests that NLVs cause sporadic gastroenteritis. These data indicate that NLV infection is a major cause of acute morbidity in military forces. Because of the limitations of available prevention and control methods, development of a vaccine against these viruses may be the best solution in the military environment. PMID- 10804154 TI - Dorsal unpaired median neurones in the insect central nervous system: towards a better understanding of the ionic mechanisms underlying spontaneous electrical activity. AB - The efferent dorsal unpaired median (DUM) neurones, which include octopaminergic neurones, are among the most intensively studied neurones in the insect central nervous system. They differ from other insect neurones in generating endogenous spontaneous overshooting action potentials. The second half of the 1980s is certain to be considered a turning point in the study of the ion channels underlying the electrical activity of DUM neurones. Recent advances made using the patch-clamp technique have stimulated an increasing interest in the understanding of the biophysical properties of both voltage-dependent and voltage independent ion channels. Patch-clamp studies of DUM neurones in cell culture demonstrate that these neurones express a wide variety of ion channels. At least five different types of K(+) channel have been identified: inward rectifier, delayed rectifier and A-like channels as well as Ca(2+)- and Na(+)-activated K(+) channels. Moreover, besides voltage-dependent Na(+) and Ca(2+)-sensitive Cl(-) channels, DUM neurones also express four types of Ca(2+) channel distinguished on the basis of their kinetics, voltage range of activation and pharmacological profile. Finally, two distinct resting Ca(2+) and Na(+) channels have been shown to be involved in maintaining the membrane potential and in regulating the firing pattern. In this review, we have also attempted critically to evaluate these existing ion channels with regard to their specific functions in the generation of the different phases of the spontaneous electrical activity of the DUM neurone. PMID- 10804155 TI - Ontogenetic changes in visual sensitivity of the parasitic salmon louse Lepeophtheirus salmonis. AB - The salmon louse, Lepeophtheirus salmonis, is an ectoparasitic copepod of salmonid fishes whose life cycle involves two broadly defined, free-living larval stages, the nauplius and the copepodid. After settling on a host, the copepodid goes through various transformations to become a mobile adult. We recorded swimming responses of free-swimming salmon lice at the naupliar, copepodid and adult stages to the onset (ON) and offset (OFF) of lights of varying spectral irradiance and polarization. Nauplii showed a prominent swim-up OFF response across the spectrum 352-652 nm, but no ON response. Copepodids exhibited a swim up ON response and a passive (sinking) OFF response across the same spectral range. Adults showed active swim-up responses to both ON and OFF stimuli, although the OFF response was proportionately stronger. The spectral range of the adult ON and OFF responses was the same as that of the copepodids and slightly greater than that of the nauplii, which did not exhibit responses at 652 nm. The absolute sensitivity of copepodids under white light (approx. 10(-13) photons m(2) s(1)) was higher than that of nauplii (approx. 10(-17) photons(-1) m(2 )s, OFF response) and that of adult female lice (approx. 10(-14) photons(-1)m(2)s). This suggests that the naupliar visual system is best suited for detection of shadows (e.g. the host) under a bright light field (daylight hours), while copepodids and adults may be more specialized for host detection at crepuscular periods and during the night, when light levels are low. None of the developmental stages responded to the rotation of the plane of polarized light or exhibited any difference in directed response when polarized light was used in place of diffuse light. PMID- 10804156 TI - Digital motion analysis as a tool for analysing the shape and performance of the circulatory system in transparent animals. AB - The analysis of perfusion parameters using the frame-to-frame technique and the observation of small blood vessels in transparent animals using video microscopy can be tedious and very difficult because of the poor contrast of the images. Injection of a fluorescent probe (fluorescein isothiocynate, FITC) bound to a high-molecular-mass dextran improved the visibility of blood vessels, but the gray-scale histogram showed blurring at the edges of the vessels. Furthermore, injection of the fluorescent probe into the ventricle of small zebrafish (Danio rerio) embryos (body mass approximately 1 mg) often resulted in reduced cardiac activity. Digital motion analysis, however, proved to be a very effective tool for analysing the shape and performance of the circulatory system in transparent animals and tissues. By subtracting the two fields of a video frame (the odd and the even frame), any movement that occurred within the 20 ms necessary for the acquisition of one field could be visualised. The length of the shifting vector generated by this subtraction, represented a direct measure of the velocity of a moving particle, i.e. an erythrocyte in the vascular system. By accumulating shifting vectors generated from several consecutive video frames, a complete trace of the routes over which erythrocytes moved could be obtained. Thus, a cast of the vascular system, except for those tiny vessels that are not entered by erythrocytes, could be obtained. Because the gray-scale value of any given pixel or any given group of pixels increased with the number of erythrocytes passing it, digital motion analysis could also be used to visualise the distribution of blood cells in transparent tissues. This method was used to describe the development of the peripheral vascular system in zebrafish larvae up to 8 days post-fertilisation. At this stage, food intake resulted in a clear redistribution of blood between muscle tissue and the gut, and alpha-adrenergic control of peripheral blood flow was established. PMID- 10804157 TI - Ventilation and gas exchange during treadmill locomotion in the American alligator (Alligator mississippiensis). AB - A number of anatomical characters of crocodilians appear to be inconsistent with their lifestyle as sit-and-wait predators. To address this paradoxical association of characters further, we measured lung ventilation and respiratory gas exchange during walking in American alligators (Alligator mississippiensis). During exercise, ventilation consisted of low-frequency, large-volume breaths. The alligators hyperventilated severely during walking with respect to their metabolic demands. Air convection requirements were among the highest and estimates of lung P(CO2) were among the lowest known in air-breathing vertebrates. Air convection requirements dropped immediately with cessation of exercise. These observations indicate that the ventilation of alligators is not limited by their locomotor movements. We suggest that the highly specialized ventilatory system of modern crocodilians represents a legacy from cursorial ancestors rather than an adaptation to a lifestyle as amphibious sit-and-wait predators. PMID- 10804158 TI - Pelvic aspiration in the American alligator (Alligator mississippiensis). AB - The pelvis of crocodilians is highly derived in that the pubic bones are isolated from the acetabulum and are attached to the ischia via moveable joints. We examined the possible role of this unusual morphology in lung ventilation by measuring ventilation, abdominal pressure and the electrical activity of several abdominal and pelvic muscles in the American alligator (Alligator mississippiensis). We found that the activity of two pelvic muscles, the ischiopubis and ischiotruncus muscles, was correlated with inspiration; these muscles rotate the pubes ventrally and thereby increase abdominal volume. During expiration, contraction of the rectus abdominis and transversus abdominis rotates the pubes dorsally. We suggest that this mechanism facilitates diaphragmatic breathing by creating space for caudal displacement of the viscera during inspiration. Because birds also use a dorso-ventral movement of the pelvis to effect ventilation, some form of pelvic aspiration may be plesiomorphic for archosaurs. PMID- 10804159 TI - Octopamine mimics the effects of parasitism on the foregut of the tobacco hornworm Manduca sexta. AB - The parasitic braconid wasp Cotesia congregata lays its eggs inside the body of the larval stage of its host, the moth Manduca sexta. The Cotesia congregata larvae develop within the hemocoel of their host until their third instar, when they emerge and spin cocoons and pupate on the outer surface of the caterpillar. From this time until their death approximately 2 weeks later, the Manduca sexta larvae show striking behavioral changes that include dramatic declines in spontaneous activity and in the time spent feeding. Coincident with these behavioral changes, it is known that octopamine titers in the hemolymph of the host become elevated by approximately 6.5-fold. Octopamine is an important modulator of neural function and behavior in insects, so we examined hosts for neural correlates to the behavioral changes that occur at parasite emergence. We found that, in addition to the changes reported earlier, after parasite emergence (post-emergence), Manduca sexta larvae also showed marked deficits in their ability to ingest food because of a disruption in the function of the frontal ganglion that results in a significant slowing or the absence of peristaltic activity in the foregut. This effect could be produced in unparasitized fifth instar larvae by application of blood from post-emergence parasitized larvae or of 10(-6)mol l(-1)d,l-octopamine (approximately the level in the hemolymph of post-emergence larvae). In contrast, blood from parasitized larvae before their parasites emerge or from unparasitized fifth-instar larvae typically had no effect on foregut activity. The effects of either post-emergence parasitized blood or 10(-6)mol l(-1) octopamine could be blocked by the octopamine antagonists phentolamine (at 10(-5)mol l(-1)) or mianserin (at 10(-7)mol l(-1)). PMID- 10804160 TI - 5-Hydroxy-tryptamine modulates pheromone-evoked local field potentials in the macroglomerular complex of the sphinx moth Manduca sexta. AB - Extra- and intracellular recordings from an intact brain preparation were used to study the effects of 5-hydroxytryptamine (5-HT or serotonin) on projection neurons in the sexually dimorphic macroglomerular complex (MGC) in the antennal lobe of the male moth Manduca sexta. The MGC is a group of three identified glomeruli specialized for synaptic processing of primary afferent information about the multi-component sex pheromone of the female. We investigated the modulatory effects of 5-HT on pheromone-evoked local field potentials in the MGC. The magnitude and duration of these potentials, which are thought to be generated by a population of pheromone-sensitive projection neurons of the MGC, were increased by 5-HT. Using intracellular recordings from the neurites of individual MGC projection neurons, we found that 5-HT increased the number of action potentials in response to pheromonal stimulation. These findings correlate well with earlier experiments that used other recording techniques. Our results are further evidence that 5-HT modulates a population of pheromone-sensitive MGC projection neurons that relay information about the pheromonal stimulus from the MGC to higher-order centers in the protocerebrum and are therefore pivotal for mate-finding and odor-guided behavior. PMID- 10804161 TI - Skin darkening, a potential social signal in subordinate arctic charr (Salvelinus alpinus): the regulatory role of brain monoamines and pro-opiomelanocortin derived peptides. AB - Arctic charr were allowed to interact in groups of three for 5 days. Skin darkness was quantified by measuring the mean brightness of individual fish before and after social interaction. Brain levels of monoamines and monoamine metabolites and plasma concentrations of cortisol, adrenocorticotropic hormone (ACTH), N-acetyl-(beta)-endorphin and alpha-melanocyte-stimulating hormone (alpha MSH) were analysed. The results show that social subordination resulted in a significant skin darkening. Furthermore, plasma concentrations of alpha-MSH, ACTH and cortisol were elevated in subordinates, and these fish also displayed elevated levels of 5-hydroxyindoleacetic acid (5-HIAA) in the telencephalon. The ratio of [5-HIAA] to serotonin [5-HT] was increased in several brain areas. In addition, the ratio of 3-methoxy-4-hydroxyphenylglycol (MHPG) to norepinephrine (NE) concentrations was significantly increased in the optic tectum of subordinate fish. Skin darkness following social interaction showed a significant positive correlation with plasma levels of alpha-MSH. Plasma levels of ACTH and alpha-MSH were both positively correlated with that of cortisol. Brain [5 HIAA]/[5-HT] ratios were positively correlated with circulating plasma levels of ACTH, and a similar positive correlation was seen between [MHPG]/[NE] ratios in the optic tectum and plasma levels of ACTH, alpha-MSH and N-acetyl-beta endorphin. In contrast, hypothalamic [MHPG]/[NE] ratios displayed a negative correlation with plasma alpha-MSH concentrations. The present study demonstrates that social stress induces skin darkening in Arctic charr and that this effect could be mediated by a stress-induced increase in the levels of alpha-MSH in the circulation. Furthermore, the results suggest that 5-HT and NE in the central nervous system could be factors regulating the pituitary release of ACTH and alpha-MSH. PMID- 10804162 TI - Object fixation by the blowfly during tethered flight in a simulated three dimensional environment. AB - The ability of flies to detect and fixate objects moving relative to their background was investigated in a flight simulator during translational tethered flight. The fly experienced optic flow that depended on its own actions and reactions in a similar way as in free-flight (closed-loop) conditions. Fixation of an object required turning responses towards it. The simulated distances between the fly, object and background were varied systematically by changing the velocities with which the object and the background pattern moved from the frontal to the back part of the fly's visual field. Fixation responses were only elicited when the object was simulated to be closer than the background. The fly's fixation performance was better with close than with more distant objects. Since, under many stimulus conditions, fixation responses were either elicited or entirely failed to be elicited, it is concluded that object fixation behaviour is gated in the visuo-motor pathway. PMID- 10804163 TI - Physiological variability in neonatal armadillo quadruplets: within- and between litter differences. AB - The role of genetics on neonatal physiological variability was examined in the nine-banded armadillo (Dasypus novemcinctus). Since armadillos give birth to only monozygous quadruplets, the genetic variation within litters is essentially zero. Quadruplets born in captivity were isolated and weighed within 8 h of birth. Oxygen consumption (V.(O2)) was measured in resting neonates by flow-through respirometry, heart rate obtained from an electrocardiogram and ventilation was measured by impedance techniques. Following the measurements, neonates were returned to the mother. Measurements were repeated at 4 and 8 days after birth. Mean heart rate significantly increased from 132 beats min(-1) on the day of birth to 169 beats min(-1) on day 8. Mean ventilation rate significantly decreased from 81 breaths min(-1) on the day of birth to 54 breaths min(-1) on day 8. During this same developmental period, mean mass significantly increased from 100 g to 129 g, and mean mass-specific oxygen consumption significantly decreased from 32.2 ml O(2)kg(-1)min(-1) to 28.6 ml O(2)kg(-1) min(-1). For all variables measured, within-litter variability was always significantly less than between-litter variability, confirming a 'sibling effect' that we attribute to the genetic components determining physiological characters. PMID- 10804164 TI - Developmental dimorphism and expression of chemosensory-mediated behavior: habitat selection by a specialist marine herbivore. AB - Developmental dimorphisms provide an opportunity to compare sensory systems and behavior patterns between different forms of a single species. Alternative morphs differing in dispersal ability often show behavioral differences that mediate life-history trade-offs. We measured the behavioral responses of both long-lived, feeding larvae and short-lived, non-feeding larvae of the specialist marine herbivore Alderia modesta during habitat selection. Larvae immediately responded to waterborne cues from the adult host algae by increasing their turning rate, by changing their swimming speed in the water and by moving in rapid hops or spiraling along the bottom. These behavior patterns retained larvae in areas where the dissolved cue was initially perceived, and prolonged exposure to the cue increased the percentage of larvae that initiated metamorphosis. Despite their differences in life span and trophic mode, both larval morphs displayed similar behavior patterns when stimulated by the waterborne cue. Long-lived larvae had a stronger response, however, suggesting that settlement behavior may offset the costs of a prolonged larval life. This is the first study to examine the effects of dimorphic development on chemosensory-mediated behavior. PMID- 10804165 TI - Horizontal flight of a swallow (Hirundo rustica) observed in a wind tunnel, with a new method for directly measuring mechanical power. AB - A swallow flying in the Lund wind tunnel was observed from the side and from behind, by two synchronised high-speed video cameras. The side-view camera provided a record of the vertical position of a white mark, applied to the feathers behind and below the eye, from which the vertical acceleration was obtained. The rear-view camera provided measurements of the mean angle of the left and right humeri above horizontal. From these data, the force acting on the body, the moment applied by each pectoralis muscle to the humerus and the rotation of the humerus were estimated and used to analyse the time course of a number of variables, including the work done by the muscles in each wing beat. The average mechanical power turned out to be more than that predicted on the basis of current estimates of body drag coefficient and profile power ratio, possibly because the bird was not flying steadily in a minimum-drag configuration. We hope to develop the method further by correlating the mechanical measurements with observations of the vortex wake and to apply it to birds that have been conditioned to hold a constant position in the test section. PMID- 10804166 TI - The functional morphology of the avian flight muscle M. Coracobrachialis posterior. AB - The extensive range of movement of the avian glenohumeral joint makes functional interpretation of any muscle that crosses the joint difficult. Multiple functional roles for the M. coracobrachialis posterior (CBP), an architecturally complex muscle that lies deep to the M. pectoralis, have been assigned on the basis of its anatomical position. The mechanical properties, neuromotor pattern during flight and the biochemical properties of the CBP in pigeons (Columba livia) were studied by in situ length/active tension and length/passive tension measurements, in vivo electromyography and muscle histochemistry. The action of the muscle was studied directly through in situ stimulation and measurement of humeral excursion in non-reduced preparations. PMID- 10804167 TI - Crustacean (malacostracan) Hox genes and the evolution of the arthropod trunk. AB - Representatives of the Insecta and the Malacostraca (higher crustaceans) have highly derived body plans subdivided into several tagma, groups of segments united by a common function and/or morphology. The tagmatization of segments in the trunk, the part of the body between head and telson, in both lineages is thought to have evolved independently from ancestors with a distinct head but a homonomous, undifferentiated trunk. In the branchiopod crustacean, Artemia franciscana, the trunk Hox genes are expressed in broad overlapping domains suggesting a conserved ancestral state (Averof, M. and Akam, M. (1995) Nature 376, 420-423). In comparison, in insects, the Antennapedia-class genes of the homeotic clusters are more regionally deployed into distinct domains where they serve to control the morphology of the different trunk segments. Thus an originally Artemia-like pattern of homeotic gene expression has apparently been modified in the insect lineage associated with and perhaps facilitating the observed pattern of tagmatization. Since insects are the only arthropods with a derived trunk tagmosis tested to date, we examined the expression patterns of the Hox genes Antp, Ubx and abd-A in the malacostracan crustacean Porcellio scaber (Oniscidae, Isopoda). We found that, unlike the pattern seen in Artemia, these genes are expressed in well-defined discrete domains coinciding with tagmatic boundaries which are distinct from those of the insects. Our observations suggest that, during the independent tagmatization in insects and malacostracan crustaceans, the homologous 'trunk' genes evolved to perform different developmental functions. We also propose that, in each lineage, the changes in Hox gene expression pattern may have been important in trunk tagmatization. PMID- 10804168 TI - Uncx4.1 is required for the formation of the pedicles and proximal ribs and acts upstream of Pax9. AB - The expression of the homeobox gene Uncx4.1 in the somite is restricted to the caudal half of the newly formed somite and sclerotome. Here we show that mice with a targeted mutation of the Uncx4.1 gene exhibit defects in the axial skeleton and ribs. In the absence of Uncx4.1, pedicles of the neural arches and proximal ribs are not formed. In addition, dorsal root ganglia are disorganized. Histological and marker analysis revealed that Uncx4.1 is not necessary for somite segmentation. It is required to maintain the condensation of the caudal half-sclerotome, from which the missing skeletal elements are derived. The loss of proximal ribs in Pax1/Pax9 double mutants and the data presented here argue for a role of Uncx4.1 upstream of Pax9 in the caudolateral sclerotome. Our results further indicate that Uncx4.1 may be involved in the differential cell adhesion properties of the somite. PMID- 10804169 TI - The paired homeobox gene Uncx4.1 specifies pedicles, transverse processes and proximal ribs of the vertebral column. AB - The axial skeleton develops from the sclerotome, a mesenchymal cell mass derived from the ventral halves of the somites, segmentally repeated units located on either side of the neural tube. Cells from the medial part of the sclerotome form the axial perichondral tube, which gives rise to vertebral bodies and intervertebral discs; the lateral regions of the sclerotome will form the vertebral arches and ribs. Mesenchymal sclerotome cells condense and differentiate into chondrocytes to form a cartilaginous pre-skeleton that is later replaced by bone tissue. Uncx4.1 is a paired type homeodomain transcription factor expressed in a dynamic pattern in the somite and sclerotome. Here we show that mice homozygous for a targeted mutation of the Uncx4.1 gene die perinatally and exhibit severe malformations of the axial skeleton. Pedicles, transverse processes and proximal ribs, elements derived from the lateral sclerotome, are lacking along the entire length of the vertebral column. The mesenchymal anlagen for these elements are formed initially, but condensation and chondrogenesis do not occur. Hence, Uncx4.1 is required for the maintenance and differentiation of particular elements of the axial skeleton. PMID- 10804170 TI - Postnatal mammary gland development requires macrophages and eosinophils. AB - Interactions between mammary epithelial and mesenchymal cells including fibroblasts and adipocytes are crucial for the proper postnatal development of the mammary ductal tree. Often overlooked, however, are the migrant cells that enter tissues at different stages of development. In this paper we identify two such cell types, macrophages and eosinophils, that are recruited around the growing terminal end buds (TEBs) during postnatal development. An important role for leukocytes in mammary gland ductal outgrowth is first demonstrated by depleting mice of leukocytes using sub-lethal (gamma)-irradiation. This treatment results in a curtailment of mammary gland epithelial development that is completely rescued by bone-marrow transplantation, concurrent with a restoration of macrophage and eosinophil recruitment around the growing ducts. Using mice homozygous for a null mutation in the gene for CSF1 (Csfm(op)/Csfm(op)), the major growth factor for macrophages, we show that in the absence of CSF1, the population of macrophages in mammary glands is depleted. In this mutant, the formation of TEBs, their outgrowth into the fat pad and the branching of the resultant ducts are all impaired. Similarly, by using mice homozygous for a null mutation in the gene for eotaxin, a major chemokine for local recruitment of eosinophils in tissue, we identify eotaxin as the necessary and sufficient chemokine responsible for eosinophil recruitment around TEBs. In the absence of eosinophils, mammary gland branch formation and to a lesser extent TEB formation are reduced. Our data show that CSF1-regulated macrophages, in collaboration with eotaxin-regulated eosinophils, have essential and complementary functions in regulating the branching morphogenesis of the mammary gland. PMID- 10804171 TI - Histone macroH2A1 is concentrated in the inactive X chromosome of female preimplantation mouse embryos. AB - MacroH2As are core histone proteins with a hybrid structure consisting of a domain that closely resembles a full-length histone H2A followed by a large nonhistone domain. We recently showed that one of the macroH2A subtypes, macroH2A1.2, is concentrated in the inactive X chromosome in adult female mammals. Here we examine the timing of the association of macroH2A1.2 with the inactive X chromosome during preimplantation mouse development in order to assess the possibility that macroH2A1 participates in the initiation of X inactivation. The association of macroH2A1.2 with one of the X chromosomes was observed in 50% of blastocysts, occurring mostly, if not exclusively, in extraembryonic cells as was expected from previous studies, which indicated that X inactivation in embryonic lineages happens after implantation. Examination of earlier embryonic stages indicates that the association of macroH2A1 with the inactive X chromosome begins between the 8- and 16-cell stages. Of the changes that are known to happen during X inactivation in preimplantation embryos, the accumulation of macroH2A1 appears to be the earliest marker of the inactive X chromosome and is the only change that has been shown to occur during the period when transcriptional silencing is initiated. PMID- 10804172 TI - brakeless is required for lamina targeting of R1-R6 axons in the Drosophila visual system. AB - Photoreceptors in the Drosophila eye project their axons retinotopically to targets in the optic lobe of the brain. The axons of photoreceptor cells R1-R6 terminate in the first optic ganglion, the lamina, while R7 and R8 axons project through the lamina to terminate in distinct layers of the second ganglion, the medulla. Here we report the identification of the gene brakeless (bks) and show that its function is required in the developing eye specifically for the lamina targeting of R1-R6 axons. In mosaic animals lacking bks function in the eye, R1 R6 axons project through the lamina to terminate in the medulla. Other aspects of visual system development appear completely normal: photoreceptor and lamina cell fates are correctly specified, R7 axons correctly target the medulla, and both correctly targeted R7 axons and mistargeted R1-R6 axons maintain their retinotopic order with respect to both anteroposterior and dorsoventral axes. bks encodes two unusually hydrophilic nuclear protein isoforms, one of which contains a putative C(2)H(2) zinc finger domain. Transgenic expression of either Bks isoform is sufficient to restore the lamina targeting of R1-R6 axons in bks mosaics, but not to retarget R7 or R8 axons to the lamina. These data demonstrate the existence of a lamina-specific targeting mechanism for R1-R6 axons in the Drosophila visual system, and provide the first entry point in the molecular characterization of this process. PMID- 10804173 TI - Hex is a transcriptional repressor that contributes to anterior identity and suppresses Spemann organiser function. AB - One of the earliest markers of anterior asymmetry in vertebrate embryos is the transcription factor Hex. We find that Hex is a transcriptional repressor that can be converted to an activator by fusing full length Hex to two copies of the minimal transcriptional activation domain of VP16 together with the flexible hinge region of the (lambda) repressor (Hex-(lambda)VP2). Retention of the entire Hex open reading frame allows one to examine Hex function without disrupting potential protein-protein interactions. Expression of Hex-(lambda)VP2 in Xenopus inhibits expression of the anterior marker Cerberus and results in anterior truncations. Such embryos have multiple notochords and disorganised muscle tissue. These effects can occur in a cell non-autonomous manner, suggesting that one role of wild-type Hex is to specify anterior structures by suppressing signals that promote dorsal mesoderm formation. In support of this idea, over expression of wild-type Hex causes cell non-autonomous dorso-anteriorization, as well as cell autonomous suppression of dorsal mesoderm. Suppression of dorsal mesoderm by Hex is accompanied by the down-regulation of Goosecoid and Chordin, while induction of dorsal mesoderm by Hex-(lambda)VP2 results in activation of these genes. Transient transfection experiments in ES cells suggest that Goosecoid is a direct target of Hex. Together, our results support a model in which Hex suppresses organiser activity and defines anterior identity. PMID- 10804174 TI - Adult insulin- and glucagon-producing cells differentiate from two independent cell lineages. AB - To analyze cell lineage in the pancreatic islets, we have irreversibly tagged all the progeny of cells through the activity of Cre recombinase. Adult glucagon alpha and insulin beta cells are shown to derive from cells that have never transcribed insulin or glucagon, respectively. Also, the beta-cell progenitors, but not alpha-cell progenitors, transcribe the pancreatic polypeptide (PP) gene. Finally, the homeodomain gene PDX1, which is expressed by adult beta-cells, is also expressed by alpha-cell progenitors. Thus the islet alpha- and beta-cell lineages appear to arise independently during ontogeny, probably from a common precursor. PMID- 10804175 TI - Hes genes regulate sequential stages of neurogenesis in the olfactory epithelium. AB - We have characterised the functions of the bHLH transcriptional repressors HES1 and HES5 in neurogenesis, using the development of the olfactory placodes in mouse embryos as a model. Hes1 and Hes5 are expressed with distinct patterns in the olfactory placodes and are subject to different regulatory mechanisms. Hes1 is expressed in a broad placodal domain, which is maintained in absence of the neural determination gene Mash1. In contrast, expression of Hes5 is restricted to clusters of neural progenitor cells and requires Mash1 function. Mutations in Hes1 and Hes5 also have distinct consequences on olfactory placode neurogenesis. Loss of Hes1 function leads both to expression of Mash1 outside of the normal domain of neurogenesis and to increased density of MASH1-positive progenitors within this domain, and results in an excess of neurons after a delay. A mutation in Hes5 does not produce any apparent defect. However, olfactory placodes that are double mutant for Hes1 and Hes5 upregulate Ngn1, a neural bHLH gene activated downstream of Mash1, and show a strong and rapid increase in neuronal density. Together, our results suggest that Hes1 regulates Mash1 transcription in the olfactory placode in two different contexts, initially as a prepattern gene defining the placodal domain undergoing neurogenesis and, subsequently, as a neurogenic gene controlling the density of neural progenitors in this domain. Hes5 synergizes with Hes1 and regulates neurogenesis at the level of Ngn1 expression. Therefore, the olfactory sensory neuron lineage is regulated at several steps by negative signals acting through different Hes genes and targeting the expression of different proneural gene homologs. PMID- 10804176 TI - The homeobox gene bozozok promotes anterior neuroectoderm formation in zebrafish through negative regulation of BMP2/4 and Wnt pathways. AB - The neuroectoderm of the vertebrate gastrula was proposed by Nieuwkoop to be regionalized into forebrain, midbrain, hindbrain and spinal cord by a two-step process. In the activation step, the Spemann gastrula organizer induces neuroectoderm with anterior character, followed by posteriorization by a transforming signal. Recently, simultaneous inhibition of BMP and Wnt signaling was shown to induce head formation in frog embryos. However, how the inhibition of BMP and Wnt signaling pathways specify a properly patterned head, and how they are regulated in vivo, is not understood. Here we demonstrate that the loss of anterior neural fates observed in zebrafish bozozok (boz) mutants occurs during gastrulation due to a reduction and subsequent posteriorization of neuroectoderm. The neural induction defect was correlated with decreased chordino expression and consequent increases in bmp2b/4 expression, and was suppressed by overexpression of BMP antagonists. Whereas expression of anterior neural markers was restored by ectopic BMP inhibition in early boz gastrulae, it was not maintained during later gastrulation. The posteriorization of neuroectoderm in boz was correlated with ectopic dorsal wnt8 expression. Overexpression of a Wnt antagonist rescued formation of the organizer and anterior neural fates in boz mutants. We propose that boz specifies formation of anterior neuroectoderm by regulating BMP and Wnt pathways in a fashion consistent with Nieuwkoop's two-step neural patterning model. boz promotes neural induction by positively regulating organizer-derived chordino and limiting the antineuralizing activity of BMP2b/4 morphogens. In addition, by negative regulation of Wnt signaling, boz promotes organizer formation and limits posteriorization of neuroectoderm in the late gastrula. PMID- 10804177 TI - The Oak Ridge Polycystic Kidney (orpk) disease gene is required for left-right axis determination. AB - Analysis of several mutations in the mouse is providing useful insights into the nature of the genes required for the establishment of the left-right axis during early development. Here we describe a new targeted allele of the mouse Tg737 gene, Tg737(Delta)2-3(beta)Gal), which causes defects in left-right asymmetry and other abnormalities during embryogenesis. The Tg737 gene was originally identified based on its association with the mouse Oak Ridge Polycystic Kidney (orpk) insertional mutation, which causes polycystic kidney disease and other defects. Complementation tests between the original orpk mutation and the new targeted knock-out mutation demonstrate that Tg737(Delta)2-3(beta)Gal) behaves as an allele of Tg737. The differences in the phenotype between the two mutations suggest that the orpk mutation is a hypomorphic allele of the Tg737 gene. Unlike the orpk allele, where all homozygotes survive to birth, embryos homozygous for the Tg737(Delta)2-3(beta)Gal) mutation arrest in development at mid-gestation and exhibit neural tube defects, enlargement of the pericardial sac and, most notably, left-right asymmetry defects. At mid-gestation the direction of heart looping is randomized, and at earlier stages in development lefty-2 and nodal, which are normally expressed asymmetrically, exhibit symmetrical expression in the mutant embryos. Additionally, we determined that the ventral node cells in mutant embryos fail to express the central cilium, which is a characteristic and potentially functional feature of these cells. The expression of both Shh and Hnf3(beta) is downregulated in the midline at E8.0, indicating that there are significant alterations in midline development in the Tg737(Delta)2-3(beta)Gal) homozygous embryos. We propose that the failure of ventral node cells to fully mature alters their ability to undergo differentiation as they migrate out of the node to contribute to the developing midline structures. Analysis of this new knockout allele allows us to define a critical role for the Tg737 gene during early embryogenesis. We have named the product of the Tg737 gene Polaris, which is based on the various polarity related defects associated with the different alleles of the Tg737 gene. PMID- 10804178 TI - Pax6 defines the di-mesencephalic boundary by repressing En1 and Pax2. AB - Transcriptional factors and signaling molecules are responsible for regionalization of the central nervous system. In the early stage of neural development, Pax6 is expressed in the prosencephalon, while En1 and Pax2 are expressed in the mesencephalon. Here, we misexpressed Pax6 in the mesencephalon to elucidate the mechanism of the di-mesencephalic boundary formation. Histological analysis, expression patterns of diencephalic marker genes, and fiber trajectory of the posterior commissure indicated that Pax6 misexpression caused a caudal shift of the di-mesencephalic boundary. Pax6 repressed En1, Pax2 and other tectum (mesencephalon)-related genes such as En2, Pax5, Pax7, but induced Tcf4, a diencephalon marker gene. To know how Pax6 represses En1 and Pax2, we ectopically expressed a dominant-active or negative form of Pax6. The dominant-active form of Pax6 showed a similar but more severe phenotype than Pax6, while the dominant-negative form showed an opposite phenotype, suggesting that Pax6 acts as a transcriptional activator. Thus Pax6 may repress tectum related genes by activating an intervening repressor. The results of misexpression experiments, together with normal expression patterns of Pax6, En1 and Pax2, suggest that repressive interaction between Pax6 and En1/Pax2 defines the di-mesencephalic boundary. PMID- 10804179 TI - Sox2 regulatory sequences direct expression of a (beta)-geo transgene to telencephalic neural stem cells and precursors of the mouse embryo, revealing regionalization of gene expression in CNS stem cells. AB - Sox2 is one of the earliest known transcription factors expressed in the developing neural tube. Although it is expressed throughout the early neuroepithelium, we show that its later expression must depend on the activity of more than one regionally restricted enhancer element. Thus, by using transgenic assays and by homologous recombination-mediated deletion, we identify a region upstream of Sox2 (-5.7 to -3.3 kb) which can not only drive expression of a (beta)-geo transgene to the developing dorsal telencephalon, but which is required to do so in the context of the endogenous gene. The critical enhancer can be further delimited to an 800 bp fragment of DNA surrounding a nuclease hypersensitive site within this region, as this is sufficient to confer telencephalic expression to a 3.3 kb fragment including the Sox2 promoter, which is otherwise inactive in the CNS. Expression of the 5.7 kb Sox2(beta)-geo transgene localizes to the neural plate and later to the telencephalic ventricular zone. We show, by in vitro clonogenic assays, that transgene expressing (and thus G418-resistant) ventricular zone cells include cells displaying functional properties of stem cells, i.e. self-renewal and multipotentiality. We further show that the majority of telencephalic stem cells express the transgene, and this expression is largely maintained over two months in culture (more than 40 cell divisions) in the absence of G418 selective pressure. In contrast, stem cells grown in parallel from the spinal cord never express the transgene, and die in G418. Expression of endogenous telencephalic genes was similarly observed in long-term cultures derived from the dorsal telencephalon, but not in spinal cord-derived cultures. Thus, neural stem cells of the midgestation embryo are endowed with region-specific gene expression (at least with respect to some networks of transcription factors, such as that driving telencephalic expression of the Sox2 transgene), which can be inherited through multiple divisions outside the embryonic environment. PMID- 10804180 TI - Relationships between extramacrochaetae and Notch signalling in Drosophila wing development. AB - The function of extramacrochaetae is required during the development of the Drosophila wing in processes such as cell proliferation and vein differentiation. extramacrochaetae encodes a transcription factor of the HLH family, but unlike other members of this family, Extramacrochaetae lacks the basic region that is involved in interaction with DNA. Some phenotypes caused by extramacrochaetae in the wing are similar to those observed when Notch signalling is compromised. Furthermore, maximal levels of extramacrochaetae expression in the wing disc are restricted to places where Notch activity is higher, suggesting that extramacrochaetae could mediate some aspects of Notch signalling during wing development. We have studied the relationships between extramacrochaetae and Notch in wing development, with emphasis on the processes of vein formation and cell proliferation. We observe strong genetic interaction between extramacrochaetae and different components of the Notch signalling pathway, suggesting a functional relationship between them. We show that the higher level of extramacrochaetae expression coincides with the domain of expression of Notch and its downstream gene Enhancer of split-m(beta). The expression of extramacrochaetae at the dorso/ventral boundary and in boundary cells between veins and interveins depends on Notch activity. We propose that at least during vein differentiation and wing margin formation, extramacrochaetae is regulated by Notch and collaborates with other Notch-downstream genes such as Enhancer of split-m(beta). PMID- 10804181 TI - Regulation of central neuron synaptic targeting by the Drosophila POU protein, Acj6. AB - Mutations in the Drosophila class IV POU domain gene, abnormal chemosensory jump 6 (acj6), have previously been shown to cause physiological deficits in odor sensitivity. However, loss of Acj6 function also has a severe detrimental effect upon coordinated larval and adult movement that cannot be explained by the simple loss in odorant detection. In addition to olfactory sensory neurons, Acj6 is expressed in a distinct subset of postmitotic interneurons in the central nervous system from late embryonic to adult stages. In the larval and adult brain, Acj6 is highly expressed in central brain, optic and antennal lobe neurons. Loss of Acj6 function in larval optic lobe neurons results in disorganized retinal axon targeting and synapse selection. Furthermore, the lamina neurons themselves exhibit disorganized synaptic arbors in the medulla of acj6 mutant pupal brains, suggesting that Acj6 may play a role in regulating synaptic connections or structure. To further test this hypothesis, we misexpressed two Acj6 isoforms in motor neurons where they are not normally found. The two Acj6 isoforms are produced from alternatively spliced acj6 transcripts, resulting in significant structural differences in the amino-terminal POU IV box. Acj6 misexpression caused marked alterations at the neuromuscular junction, with contrasting effects upon nerve terminal branching and synapse formation associated with specific Acj6 isoforms. Our results suggest that the class IV POU domain factor, Acj6, may play an important role in regulating synaptic target selection by central neurons and that the amino-terminal POU IV box is important for regulation of Acj6 activity. PMID- 10804182 TI - The capacitating agent bicarbonate induces protein kinase A-dependent changes in phospholipid transbilayer behavior in the sperm plasma membrane. AB - A flow cytometric procedure was used to follow the effect of bicarbonate, a key inducer of sperm capacitation in vitro, on the transbilayer behavior of C6NBD phospholipids in the plasma membrane of living acrosome-intact boar spermatozoa under physiological conditions. In the absence of bicarbonate, 97% of C6NBD phosphatidylserine and 78% of C6NBD-phosphatidylethanolamine was rapidly translocated from the outer leaflet to the inner, whereas relatively little C6NBD phosphatidylcholine and C6NBD-sphingomyelin was translocated (15% and 5%, respectively). Inclusion of 15 mM bicarbonate/5%CO(2) markedly slowed down the rates of translocation of the aminophospholipids without altering their final distribution, whereas it increased the proportions of C6NBD-phosphatidylcholine and C6NBD-sphingomyelin translocated (30% and 20%, respectively). Bicarbonate activated very markedly the outward translocation of all four phospholipid classes. The changes in C6NBD-phospholipid behavior were accompanied by increased membrane lipid disorder as detected by merocyanine 540, and also by increased potential for phospholipase catabolism of the C6NBD-phospholipid probes. All three changes were mediated via a cAMP-dependent protein phosphorylation pathway. We suspect that the changes result from an activation of the non- specific bidirectional translocase ('scramblase'). They have important implications with respect to sperm fertilizing function. PMID- 10804183 TI - Activation of the Notch pathway in the hair cortex leads to aberrant differentiation of the adjacent hair-shaft layers. AB - Little is known about the mechanisms underlying the generation of various cell types in the hair follicle. To investigate the role of the Notch pathway in this process, transgenic mice were generated in which an active form of Notch1 (Notch(DeltaE)) was overexpressed under the control of the mouse hair keratin A1 (MHKA1) promoter. MHKA-Notch(DeltaE) is expressed only in one precursor cell type of the hair follicle, the cortex. Transgenic mice could be easily identified by the phenotypes of curly whiskers and wavy, sheen pelage hair. No effects of activated Notch on proliferation were detected in hair follicles of the transgenic mice. We find that activating Notch signaling in the cortex caused abnormal differentiation of the medulla and the cuticle, two neighboring cell types that did not express activated Notch. We demonstrate that these non autonomous effects are likely caused by cell-cell interactions between keratinocytes within the hair follicle and that Notch may function in such interactions either by directing the differentiation of follicular cells or assisting cells in interpreting a gradient emanating from the dermal papilla. PMID- 10804184 TI - The homeobox gene Hex is required in definitive endodermal tissues for normal forebrain, liver and thyroid formation. AB - The homeobox gene Hex is expressed in the anterior visceral endoderm (AVE) and rostral definitive endoderm of early mouse embryos. Later, Hex transcripts are detected in liver, thyroid and endothelial precursor cells. A null mutation was introduced into the Hex locus by homologous recombination in embryonic stem cells. Hex mutant embryos exhibit varying degrees of anterior truncation as well as liver and thyroid dysplasia. The liver diverticulum is formed but migration of hepatocytes into the septum transversum fails to occur. Development of the thyroid is arrested at the thyroid bud stage at 9.5 dpc. Brain defects are restricted to the rostral forebrain and have a caudal limit at the zona limitans intrathalamica, the boundary between dorsal and ventral thalamus. Analysis of Hex(-/-) mutants at early stages shows that the prospective forebrain ectoderm is correctly induced and patterned at 7.5 days post coitum (dpc), but subsequently fails to develop. AVE markers are expressed and correctly positioned but development of rostral definitive endoderm is greatly disturbed in Hex(-/-) embryos. Chimeric embryos composed of Hex(-/-) cells developing within a wild type visceral endoderm show forebrain defects indicating that Hex is required in the definitive endoderm. All together, these results demonstrate that Hex function is essential in definitive endoderm for normal development of the forebrain, liver and thyroid gland. PMID- 10804185 TI - A transitional stage in the commitment of mesoderm to hematopoiesis requiring the transcription factor SCL/tal-1. AB - In this report, we describe the identification and characterization of an early embryoid body-derived colony, termed the transitional colony, which contains cell populations undergoing the commitment of mesoderm to the hematopoietic and endothelial lineages. Analysis of individual transitional colonies indicated that they express Brachyury as well as flk-1, SCL/tal-1, GATA-1, (beta)H1 and (beta)major reflecting the combination of mesodermal, hematopoietic and endothelial populations. This pattern differs from that found in the previously described hemangioblast-derived blast cell colonies in that they typically lacked Brachyury expression, consistent with their post-mesodermal stage of development (Kennedy, M., Firpo, M., Choi, K., Wall, C., Robertson, S., Kabrun, N. and Keller, G. (1997) Nature 386, 488-493). Replating studies demonstrated that transitional colonies contain low numbers of primitive erythroid precursors as well as a subset of precursors associated with early stage definitive hematopoiesis. Blast cell colonies contain higher numbers and a broader spectrum of definitive precursors than found in the transitional colonies. ES cells homozygous null for the SCL/tal-1 gene, a transcription factor known to be essential for development of the primitive and definitive hematopoietic systems, were not able to form blast colonies but did form transitional colonies. Together these findings suggest that the transitional colony represents a stage of development earlier than the blast cell colony and one that uniquely defines the requirement for a functional SCL/tal-1 gene for the progression to hematopoietic commitment. PMID- 10804186 TI - The bHLH transcription factor dHAND controls Sonic hedgehog expression and establishment of the zone of polarizing activity during limb development. AB - Limb outgrowth and patterning of skeletal elements are dependent on complex tissue interactions involving the zone of polarizing activity (ZPA) in the posterior region of the limb bud and the apical ectodermal ridge. The peptide morphogen Sonic hedgehog (SHH) is expressed specifically in the ZPA and, when expressed ectopically, is sufficient to mimic its functions, inducing tissue growth and formation of posterior skeletal elements. We show that the basic helix loop-helix transcription factor dHAND is expressed posteriorly in the developing limb prior to Shh and subsequently occupies a broad domain that encompasses the Shh expression domain. In mouse embryos homozygous for a dHAND null allele, limb buds are severely underdeveloped and Shh is not expressed. Conversely, misexpression of dHAND in the anterior region of the limb bud of transgenic mice results in formation of an additional ZPA, revealed by ectopic expression of Shh and its target genes, and resulting limb abnormalities that include preaxial polydactyly with duplication of posterior skeletal elements. Analysis of mouse mutants in which Hedgehog expression is altered also revealed a feedback mechanism in which Hedgehog signaling is required to maintain the full dHAND expression domain in the developing limb. Together, these findings identify dHAND as an upstream activator of Shh expression and important transcriptional regulator of limb development. PMID- 10804187 TI - Molecular analysis of external genitalia formation: the role of fibroblast growth factor (Fgf) genes during genital tubercle formation. AB - The molecular mechanisms underlying the development of the external genitalia in mammals have been very little examined. Recent gene knockout studies have suggested that the developmental processes of its anlage, the genital tubercle (GT), have much in common with those of limb buds. The Fgf genes have been postulated as regulating several downstream genes during organogenesis. Fgf8 was expressed in the distal urethral plate epithelium of the genital tubercle (GT) together with other markers such as the Msx1, Fgf10, Hoxd13 and Bmp4 expressed in the mesenchyme. To analyze the role of the FGF system during GT formation, an in vitro organ culture system was utilized. It is suggested that the distal urethral plate epithelium of GT, the Fgf8-expressing region, regulates the outgrowth of GT. Ectopic application of FGF8 beads to the murine GT induced mesenchymal gene expression, and also promoted the outgrowth of the GT. Experiments utilizing anti FGF neutralizing antibody suggested a growth-promoting role for FGF protein(s) in GT outgrowth. In contrast, despite its vital role during limb-bud formation, Fgf10 appears not to be primarily essential for initial outgrowth of GT, as extrapolated from Fgf10(-/-) GTs. However, the abnormal external genitalia development of Fgf10(-/-) perinatal mice suggested the importance of Fgf10 in the development of the glans penis and the glans clitoridis. These results suggest that the FGF system is a key element in orchestrating GT development. PMID- 10804188 TI - aph-2 encodes a novel extracellular protein required for GLP-1-mediated signaling. AB - In animal development, numerous cell-cell interactions are mediated by the GLP 1/LIN-12/NOTCH family of transmembrane receptors. These proteins function in a signaling pathway that appears to be conserved from nematodes to humans. We show here that the aph-2 gene is a new component of the GLP-1 signaling pathway in the early Caenorhabditis elegans embryo, and that proteins with sequence similarity to the APH-2 protein are found in Drosophila and vertebrates. During the GLP-1 mediated cell interactions in the C. elegans embryo, APH-2 is associated with the cell surfaces of both the signaling, and the responding, blastomeres. Analysis of chimeric embryos that are composed of aph-2(+) and aph-2(-) blastomeres suggests that aph-2(+) function may be provided by either the signaling or responding blastomere. PMID- 10804189 TI - Parent-of-origin effects on seed development in Arabidopsis thaliana require DNA methylation. AB - Some genes in mammals and flowering plants are subject to parental imprinting, a process by which differential epigenetic marks are imposed on male and female gametes so that one set of alleles is silenced on chromosomes contributed by the mother while another is silenced on paternal chromosomes. Therefore, each genome contributes a different set of active alleles to the offspring, which develop abnormally if the parental genome balance is disturbed. In Arabidopsis, seeds inheriting extra maternal genomes show distinctive phenotypes such as low weight and inhibition of mitosis in the endosperm, while extra paternal genomes result in reciprocal phenotypes such as high weight and endosperm overproliferation. DNA methylation is known to be an essential component of the parental imprinting mechanism in mammals, but there is less evidence for this in plants. For the present study, seed development was examined in crosses using a transgenic Arabidopsis line with reduced DNA methylation. Crosses between hypomethylated and wild-type diploid plants produced similar seed phenotypes to crosses between plants with normal methylation but different ploidies. This is consistent with a model in which hypomethylation of one parental genome prevents silencing of alleles that would normally be active only when inherited from the other parent - thus phenocopying the effects of extra genomes. These results suggest an important role for methylation in parent-of-origin effects, and by inference parental imprinting, in plants. The phenotype of biparentally hypomethylated seeds is less extreme than the reciprocal phenotypes of uniparentally hypomethylated seeds. The observation that development is less severely affected if gametes of both sexes (rather than just one) are 'neutralized' with respect to parent-of-origin effects supports the hypothesis that parental imprinting is not necessary to regulate development. PMID- 10804190 TI - Activin/nodal responsiveness and asymmetric expression of a Xenopus nodal-related gene converge on a FAST-regulated module in intron 1. AB - Vertebrate Nodal-related factors play central roles in mesendoderm induction and left-right axis specification, but the mechanisms regulating their expression are largely unknown. We identify an element in Xnr1 intron 1 that is activated by activin and Vg1, autoactivated by Xnrs, and suppressed by ventral inducers like BMP4. Intron 1 contains three FAST binding sites on which FAST/Smad transcriptional complexes can assemble; these sites are differentially involved in intron 1-mediated reporter gene expression. Interference with FAST function abolishes intron 1 activity, and transcriptional activation of Xnrs by activin in embryonic tissue explant assays, identifying FAST as an essential mediator of Xnr autoregulation and/or 'signal relay' from activin-like molecules. Furthermore, the mapping of endogenous activators of the Xnr1 intronic enhancer within Xenopus embryos agrees well with the pattern of Xnr1 transcription during embryogenesis. In transgenic mice, Xnr1 intron 1 mimics a similarly located enhancer in the mouse nodal gene, and directs FAST site-dependent expression in the primitive streak during gastrulation, and unilateral expression during early somitogenesis. The FAST cassette is similar in an ascidian nodal-related gene, suggesting an ancient origin for this regulatory module. Thus, an evolutionarily conserved intronic enhancer in Xnr1 is involved in both mesendoderm induction and asymmetric expression during left-right axis formation. PMID- 10804191 TI - Retroviral inhibition of cAMP-dependent protein kinase inhibits myelination but not Schwann cell mitosis stimulated by interaction with neurons. AB - Schwann cells are the myelinating glia of the peripheral nervous system. Neuron Schwann cell contact profoundly affects several aspects of Schwann cell phenotype, including stimulation of mitosis and myelin formation. Many reports suggest that neuronal contact exerts this influence on Schwann cells by elevating Schwann cell cAMP and activating cAMP-dependent protein kinase A (PKA). To elucidate the importance of Schwann cell PKA in neuronal stimulation of Schwann cell mitosis and myelination, the gene encoding the PKA inhibitory protein RIalphaAB or PKIEGFP was delivered to Schwann cells using retroviral vectors. PKA inhibitory retroviral vectors effectively blocked forskolin-stimulated Schwann cell mitosis and morphological change, demonstrating the ability of the vectors to inhibit PKA in infected Schwann cells. Treatment of dorsal root ganglia neuron Schwann cell cocultures with H-89 (10 microm) or KT5720 (1-10 microm), chemical inhibitors selective for PKA, significantly inhibited neuronal stimulation of Schwann cell mitosis. In contrast, retrovirus-mediated inhibition of Schwann cell PKA had no effect on the ability of neurons to stimulate Schwann cell mitosis. However, markedly fewer myelin segments were formed by Schwann cells expressing PKA inhibitory proteins compared with controls. These results suggest that activation of Schwann cell PKA is required for myelin formation but not for Schwann cell mitosis stimulated by interaction with neurons. PMID- 10804192 TI - Hypoxia-induced silencing of NMDA receptors in turtle neurons. AB - Hypoxia-induced suppression of NMDA receptors (NMDARs) in western painted turtle (Chrysemys picta) cortical neurons may be critical for surviving months of anoxic dormancy. We report that NMDARs are silenced by at least three different mechanisms operating at different times during anoxia. In pyramidal neurons from cerebrocortex, 1-8 min anoxia suppressed NMDAR activity (Ca(2+) influx and open probability) by 50-60%. This rapid decrease in receptor activity was controlled by activation of phosphatase 1 or 2A but was not associated with an increase in [Ca(2+)](i). However, during 2 hr of anoxia, [Ca(2+)](i) in cerebrocortical neurons increased by 35%, and suppression of NMDARs was predicted by the increase of [Ca(2+)](i) and controlled by calmodulin. An additional mechanism of NMDAR silencing, reversible removal of receptors from the cell membrane, was found in cerebrocortex of turtles remaining anoxic at 3 degrees C for 3-21 d. When suppression of NMDARs was prevented with phosphatase inhibitors, tolerance of anoxia was lost. Silencing of NMDARs is thus critical to the remarkable ability of C. picta to tolerate life without oxygen. PMID- 10804193 TI - Developmentally regulated NMDA receptor-dependent dephosphorylation of cAMP response element-binding protein (CREB) in hippocampal neurons. AB - Developmental changes in the signaling properties of NMDA receptors have been proposed to underlie the loss of plasticity that accompanies brain maturation. Calcium influx through postsynaptic NMDA receptors can stimulate neuronal gene expression via signaling pathways such as the Ras-MAP kinase (MAPK) pathway and the transcription factor cAMP response element-binding protein (CREB). We analyzed MAPK (Erk1/2) and CREB activation in response to NMDA receptor stimulation during the development of hippocampal neurons in culture. At all stages of development NMDA stimulation induced a rapid phosphorylation of CREB on Ser-133 (phospho-CREB). However, the time course of decline in phospho-CREB changed dramatically with neuronal maturation. At 7 d in vitro (7 DIV) phospho CREB remained elevated 2 hr after strong NMDA stimulation, whereas at 14 DIV phospho-CREB rose only transiently and fell back to below basal levels within 30 min. Moreover, at 14 DIV, but not at 7 DIV, NMDA receptor stimulation induced a dephosphorylation of CREB that previously had been phosphorylated by KCl depolarization or forskolin, suggesting an NMDA receptor-dependent activation of a CREB phosphatase. There was no developmental change in the time course of phospho-CREB induction that followed KCl depolarization or PKA activation, nor was there a developmental change in the time course of phospho-Erk1/2 induced by NMDA receptor activation. We suggest that, during neuronal maturation, NMDA receptor activation becomes linked specifically to protein phosphatases that act on Ser-133 of CREB. Such a developmentally regulated switch in the mode of NMDA receptor coupling to intracellular signaling pathways may contribute to the changes in neural plasticity observed during brain development. PMID- 10804194 TI - Protein phosphatase-1 regulation in the induction of long-term potentiation: heterogeneous molecular mechanisms. AB - Protein phosphatase inhibitor-1 (I-1) has been proposed as a regulatory element in the signal transduction cascade that couples postsynaptic calcium influx to long-term changes in synaptic strength. We have evaluated this model using mice lacking I-1. Recordings made in slices prepared from mutant animals and also in anesthetized mutant animals indicated that long-term potentiation (LTP) is deficient at perforant path-dentate granule cell synapses. In vitro, this deficit was restricted to synapses of the lateral perforant path. LTP at Schaffer collateral-CA1 pyramidal cell synapses remained normal. Thus, protein phosphatase 1-mediated regulation of NMDA receptor-dependent synaptic plasticity involves heterogeneous molecular mechanisms, in both different dendritic subregions and different neuronal subtypes. Examination of the performance of I-1 mutants in spatial learning tests indicated that intact LTP at lateral perforant path granule cell synapses is either redundant or is not involved in this form of learning. PMID- 10804195 TI - Differential expression of group I metabotropic glutamate receptors in functionally distinct hippocampal interneurons. AB - Metabotropic glutamate receptors (mGluRs) have been proposed to be involved in oscillatory rhythmic activity in the hippocampus. However, the subtypes of mGluRs involved and their precise distribution in different populations of interneurons is unclear. In this study, we combined functional analysis of mGluR-mediated inward currents in CA1 oriens-alveus interneurons with anatomical and immunocytochemical identification of these interneurons and expression analysis of group I mGluR using single-cell reverse transcription-PCR (RT-PCR). Four major interneuron subtypes could be distinguished based on the mGluR-mediated inward current induced by the application of 100 microm trans-(1S,3R)-1 aminocyclopentane-1, 3-dicarboxylic acid (ACPD) under voltage-clamp conditions and the action potential firing pattern under current-clamp conditions. Type I interneurons responded with a large inward current of approximately 224 pA, were positive for somatostatin, and the majority expressed both mGluR1 and mGluR5. Type II interneurons responded with an inward current of approximately 80 pA, contained calbindin, and expressed mainly mGluR1. Type III interneurons responded with an inward current of approximately 60 pA. These interneurons were fast spiking, contained parvalbumin, and expressed mainly mGluR5. Type IV interneurons did not respond with an inward current upon application of ACPD, yet they expressed group I mGluRs. Activation of group I mGluRs under current-clamp conditions increased spike frequency and resulted in rhythmic firing activity in type I and II, but not in type III and IV, interneurons. RT-PCR results suggest that activation of mGluR1 in the subsets of GABAergic interneurons, classified here as type I and II, may play an important role in mediating synchronous activity. PMID- 10804196 TI - The mechanisms of hsp27 antibody-mediated apoptosis in retinal neuronal cells. AB - Although elevated titers of serum antibodies to hsp27 accompany human diseases such as cancer and glaucoma, evidence of their pathogenic effects is lacking. Here we present novel evidence that exogenously applied hsp27 antibody enters neuronal cells in human retina by an endocytic mechanism. Subsequent to internalization, hsp27 antibody facilitates apoptotic cell death as characterized by morphological assessment, DNA fragmentation, and the activation of cysteine aspartic acid proteases. In addition, we demonstrate that after internalization, hsp27 antibody is detected in discrete cytoplasmic and nuclear structures and colocalizes to actin cytoskeleton. Hsp27 antibody binding to actin results in depolymerization and proteolytic cleavage of actin in a dose-dependent manner. These results suggest that exogenous hsp27 antibody may induce neuronal apoptosis by inactivating or attenuating the ability of native hsp27 to stabilize actin cytoskeleton, thereby providing a novel mechanism by which autoantibodies to hsp27 may impair cell survival in selective human diseases. PMID- 10804197 TI - A novel nervous system beta subunit that downregulates human large conductance calcium-dependent potassium channels. AB - The pore-forming alpha subunits of many ion channels are associated with auxiliary subunits that influence channel expression, targeting, and function. Several different auxiliary (beta) subunits for large conductance calcium dependent potassium channels of the Slowpoke family have been reported, but none of these beta subunits is expressed extensively in the nervous system. We describe here the cloning and functional characterization of a novel Slowpoke beta4 auxiliary subunit in human and mouse, which exhibits only limited sequence homology with other beta subunits. This beta4 subunit coimmunoprecipitates with human and mouse Slowpoke. beta4 is expressed highly in human and monkey brain in a pattern that overlaps strikingly with Slowpoke alpha subunit, but in contrast to other Slowpoke beta subunits, it is expressed little (if at all) outside the nervous system. Also in contrast to other beta subunits, beta4 downregulates Slowpoke channel activity by shifting its activation range to more depolarized voltages and slowing its activation kinetics. beta4 may be important for the critical roles played by Slowpoke channels in the regulation of neuronal excitability and neurotransmitter release. PMID- 10804198 TI - Pregnenolone sulfate modulates inhibitory synaptic transmission by enhancing GABA(A) receptor desensitization. AB - We examined the effects of the neurosteroid pregnenolone sulfate (PS) on GABA(A) receptor-mediated synaptic currents and currents elicited by rapid applications of GABA onto nucleated outside-out patches in cultured postnatal rat hippocampal neurons. At 10 microm, PS significantly depressed peak responses and accelerated the decay of evoked inhibitory synaptic currents. In nucleated outside-out patches, PS depressed peak currents and speeded deactivation after 5 msec applications of a saturating concentration of GABA. PS also increased the rate and degree of macroscopic GABA receptor desensitization during prolonged GABA applications. In a paired GABA application paradigm, PS slowed the rate of recovery from desensitization. In contrast to its prominent effects on currents produced by saturating GABA concentrations, PS had only small effects on peak currents and failed to alter deactivation after brief applications of the weakly desensitizing GABA(A) receptor agonists taurine and beta-alanine. However, when beta-alanine was applied for a sufficient duration to promote receptor desensitization, PS augmented macroscopic desensitization and slowed deactivation. These results suggest that PS inhibits GABA-gated chloride currents by enhancing receptor desensitization and stabilizing desensitized states. This contention is supported by kinetic modeling studies in which increases in the rate of entry into doubly liganded desensitized states mimic most effects of PS. PMID- 10804199 TI - An intramolecular interaction between Src homology 3 domain and guanylate kinase like domain required for channel clustering by postsynaptic density-95/SAP90. AB - Members of the postsynaptic density-95 (PSD-95)/SAP90 family of membrane associated guanylate kinase (MAGUK) proteins function as multimodular scaffolds that organize protein-signaling complexes at neuronal synapses. MAGUK proteins contain PDZ, Src homology 3 (SH3), and guanylate kinase (GK)-like domains, all of which can function as sites for specific protein-protein interactions. We report here a direct protein-protein interaction between the SH3 domain and the GK region in the PSD-95 family of MAGUKs. The SH3 domain of the PSD-95 family appears to have an atypical binding specificity, because the classical SH3 binding (-P-X-X-P-) motif is absent from the GK domain. Although SH3-GK binding can occur in either an intramolecular or intermolecular manner, the intramolecular mode is preferred, possibly because of additional tertiary interactions available when the SH3 and GK domains are adjacent in the same polypeptide. Mutations disrupting the intramolecular SH3-GK interaction do not interfere with PSD-95 association with the K(+) channel Kv1.4 or with the GK domain-binding protein GKAP. The same mutations, however, inhibit the clustering of Kv1.4 by PSD-95, suggesting that the intramolecular SH3-GK interaction may modulate the clustering activity of PSD-95. PMID- 10804200 TI - GABA(A) receptor epsilon and theta subunits display unusual structural variation between species and are enriched in the rat locus ceruleus. AB - Previously, GABA(A) receptor epsilon and theta subunits have been identified only in human. Here, we describe properties of the epsilon and theta subunit genes from mouse and rat that reveal an unusually high level of divergence from their human homologs. In addition to a low level of amino acid sequence conservation ( approximately 70%), the rodent epsilon subunit cDNAs encode a unique Pro/Glx motif of approximately 400 residues within the N-terminal extracellular domain of the subunits. Transcripts of the rat epsilon subunit were detected in brain and heart, whereas the mouse theta subunit mRNA was detectable in brain, lung, and spleen by Northern blot analysis. In situ hybridization revealed a particularly strong signal for both subunit mRNAs in rat locus ceruleus in which expression was detectable from the first postnatal day. Lower levels of coexpression were also detected in other brainstem nuclei and in the hypothalamus. However, the expression pattern of theta subunit mRNA was more widespread than that of epsilon subunit, being found also in the cerebral cortex of rat pups. In contrast to primate brain, neither subunit was expressed in the hippocampus or substantia nigra. The results indicate that GABA(A) receptor epsilon and theta subunits are evolving at a much faster rate than other known GABA(A) receptor subunits and that their expression patterns and functional properties may differ significantly between species. PMID- 10804201 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP), a neuron-derived peptide regulating glial glutamate transport and metabolism. AB - In the brain, glutamatergic neurotransmission is terminated predominantly by the rapid uptake of synaptically released glutamate into astrocytes through the Na(+) dependent glutamate transporters GLT-1 and GLAST and its subsequent conversion into glutamine by the enzyme glutamine synthetase (GS). To date, several factors have been identified that rapidly alter glial glutamate uptake by post translational modification of glutamate transporters. The only condition known to affect the expression of glial glutamate transporters and GS is the coculturing of glia with neurons. We now demonstrate that neurons regulate glial glutamate turnover via pituitary adenylate cyclase-activating polypeptide (PACAP). In the cerebral cortex PACAP is synthesized by neurons and acts on the subpopulation of astroglia involved in glutamate turnover. Exposure of astroglia to PACAP increased the maximal velocity of [(3)H]glutamate uptake by promoting the expression of GLT-1, GLAST, and GS. Moreover, the stimulatory effects of neuron conditioned medium on glial glutamate transporter expression were attenuated in the presence of PACAP-inactivating antibodies or the PACAP receptor antagonist PACAP 6-38. In contrast to PACAP, vasoactive intestinal peptide promoted glutamate transporter expression only at distinctly higher concentrations, suggesting that PACAP exerts its effects on glial glutamate turnover via PAC1 receptors. Although PAC1 receptor-dependent activation of protein kinase A (PKA) was sufficient to promote the expression of GLAST, the activation of both PKA and protein kinase C (PKC) was required to promote GLT-1 expression optimally. Given the existence of various PAC1 receptor isoforms that activate PKA and PKC to different levels, these findings point to a complex mechanism by which PACAP regulates glial glutamate transport and metabolism. Disturbances of these regulatory mechanisms could represent a major cause for glutamate-associated neurological and psychiatric disorders. PMID- 10804202 TI - Evidence for seeding of beta -amyloid by intracerebral infusion of Alzheimer brain extracts in beta -amyloid precursor protein-transgenic mice. AB - Many neurodegenerative diseases are associated with the abnormal sequestration of disease-specific proteins in the brain, but the events that initiate this process remain unclear. To determine whether the deposition of the beta-amyloid peptide (Abeta), a key pathological feature of Alzheimer's disease (AD), can be induced in vivo, we infused dilute supernatants of autopsy-derived neocortical homogenates from Alzheimer's patients unilaterally into the hippocampus and neocortex of 3-month-old beta-amyloid precursor protein (betaAPP)-transgenic mice. Up to 4 weeks after the infusion there was no Abeta-deposition in the brain; however, after 5 months, the AD-tissue-injected hemisphere of the transgenic mice had developed profuse Abeta-immunoreactive senile plaques and vascular deposits, some of which were birefringent with Congo Red. There was limited deposition of diffuse Abeta also in the brains of betaAPP-transgenic mice infused with tissue from an age-matched, non-AD brain with mild beta-amyloidosis, but none in mice receiving extract from a young control case. Abeta deposits also were not found in either vehicle-injected or uninjected transgenic mice or in any nontransgenic mice. The results show that cerebral beta-amyloid can be seeded in vivo by a single inoculation of dilute AD brain extract, demonstrating a key pathogenic commonality between beta-amyloidosis and other neurodegenerative diseases involving abnormal protein polymerization. The paradigm can be used to clarify the conditions that initiate in vivo beta-amyloidogenesis in the brain and may yield a more authentic animal model of Alzheimer's disease and other neurodegenerative disorders. PMID- 10804203 TI - IFNgamma enhances microglial reactions to hippocampal axonal degeneration. AB - Glial reactivity is implicated in CNS repair and regenerative responses. Microglia, the cells responding earliest to axonal injury, produce tumor necrosis factor-alpha (TNFalpha), a cytokine with both cytopathic and neuroprotective effects. We have studied activation of hippocampal microglia to produce TNFalpha in response to transection of perforant path axons in SJL/J mice. TNFalpha mRNA was produced in a transient manner, peaking at 2 d and falling again by 5 d after lesioning. This was unlike other markers of glial reactivity, such as Mac-1 upregulation, which were sustained over longer time periods. Message for the immune cytokine interferon-gamma (IFNgamma) was undetectable, and glial reactivity to axonal lesions occurred as normal in IFNgamma-deficient mice. Microglial responses to lesion-induced neuronal injury were markedly enhanced in myelin basic protein promoter-driven transgenic mice, in which IFNgamma was endogenously produced in hippocampus. The kinetics of TNFalpha downregulation 5 d after lesion was not affected by transgenic IFNgamma, indicating that IFNgamma acts as an amplifier and not an inducer of response. These results are discussed in the context of a regenerative role for TNFalpha in the CNS, which is innately regulated and potentiated by IFNgamma. PMID- 10804204 TI - Vasoactive intestinal peptide and pituitary adenylyl cyclase-activating polypeptide inhibit tumor necrosis factor-alpha production in injured spinal cord and in activated microglia via a cAMP-dependent pathway. AB - Tumor necrosis factor-alpha (TNF-alpha) production accompanies CNS insults of all kinds. Because the neuropeptide vasoactive intestinal peptide (VIP) and the structurally related peptide pituitary adenylyl cyclase-activating polypeptide (PACAP) have potent anti-inflammatory effects in the periphery, we investigated whether these effects extend to the CNS. TNF-alpha mRNA was induced within 2 hr after rat spinal cord transection, and its upregulation was suppressed by a synthetic VIP receptor agonist. Cultured rat microglia were used to examine the mechanisms underlying this inhibition because microglia are the likely source of TNF-alpha in injured CNS. In culture, increases in TNF-alpha mRNA resulting from lipopolysaccharide (LPS) stimulation were reduced significantly by 10(-7) m VIP and completely eliminated by PACAP at the same concentration. TNF-alpha protein levels were reduced 90% by VIP or PACAP at 10(-7) m. An antagonist of VPAC(1) receptors blocked the action of VIP and PACAP, and a PAC(1) antagonist blocked the action of PACAP. A direct demonstration of VIP binding on microglia and the existence of mRNAs for VPAC(1) and PAC(1) (but not VPAC(2)) receptors argue for a receptor-mediated effect. The action of VIP is cAMP-mediated because (1) activation of cAMP by forskolin mimics the action; (2) PKA inhibition by H89 reverses the neuropeptide-induced inhibition; and (3) the lipophilic neuropeptide mimic, stearyl-norleucine(17) VIP (SNV), which does not use a cAMP-mediated pathway, fails to duplicate the inhibition. We conclude that VIP and PACAP inhibit the production of TNF-alpha from activated microglia by a cAMP-dependent pathway. PMID- 10804205 TI - N-CAM binding inhibits the proliferation of hippocampal progenitor cells and promotes their differentiation to a neuronal phenotype. AB - Cell adhesion molecules (CAMs) play important roles during the development of the nervous system. On the basis of our previous observations that binding of the neural CAM (N-CAM) inhibits astrocyte proliferation and alters gene expression, we hypothesized that N-CAM may influence the balance between the proliferation and the differentiation of neural progenitor cells. Rat and mouse hippocampal progenitor cells were cultured and showed dependence on basic FGF for proliferation, immunoreactivity for nestin, the presence of limited numbers of differentiated cells, and the ability to generate glial cells and neurons under different culture conditions. Addition of soluble N-CAM reduced cell proliferation in a dose-dependent manner with no evidence of apoptosis. The inhibition of proliferation by N-CAM was accompanied by an induction of differentiation to the neuronal lineage, as indicated by a twofold increase in the percentage of microtubule-associated protein 2-positive cells even in the presence of mitogenic growth factors. Experiments using hippocampal cells from N CAM knock-out mice indicated that N-CAM on the cell surface is not required for these effects, suggesting the existence of heterophilic signaling. These results support a role for N-CAM and N-CAM ligands in the inhibition of proliferation and the induction of neural differentiation of hippocampal neural progenitor cells. PMID- 10804206 TI - Caspase-mediated degradation of AMPA receptor subunits: a mechanism for preventing excitotoxic necrosis and ensuring apoptosis. AB - Activation of ionotropic glutamate receptors of the AMPA and NMDA subtypes likely contributes to neuronal injury and death in various neurodegenerative disorders. Excitotoxicity can manifest as either apoptosis or necrosis, but the mechanisms that determine the mode of cell death are not known. We now report that levels of AMPA receptor subunits GluR-1 and GluR-4 are rapidly decreased in cultured rat hippocampal neurons undergoing apoptosis in response to withdrawal of trophic support (WTS), whereas levels of NMDA receptor subunits NR1, NR2A, and NR2B are unchanged. Exposure of isolated synaptosomal membranes to "apoptotic" cytosolic extracts resulted in rapid degradation of AMPA receptor subunits. Treatment of cells and synaptosomal membranes with the caspase inhibitors prevented degradation of AMPA receptor subunits, demonstrating a requirement for caspases in the process. Calcium responses to AMPA receptor activation were reduced after withdrawal of trophic support and enhanced after treatment with caspase inhibitors. Vulnerability of neurons to excitotoxic necrosis was decreased after withdrawal of trophic support and potentiated by treatment with caspase inhibitors. Our data indicate that caspase-mediated degradation of AMPA receptor subunits occurs during early periods of cell stress and may serve to ensure apoptosis by preventing excitotoxic necrosis. PMID- 10804207 TI - Morphology and growth patterns of developing thalamocortical axons. AB - It is increasingly evident that the actions of guidance factors depend critically on the cellular and molecular context in which they operate. For this reason we examined the growth cone morphology and behavior of thalamic fibers in the relatively natural environment of a slice preparation containing the entire pathway from thalamus to cortex. Axons were labeled with DiI crystals and imaged with a laser-scanning confocal microscope for up to 8 hr. Their behavior was analyzed in terms of morphology, extension rates, shape of trajectory, frequency of branching, and percentage of time spent in advance, pause, and retraction. Thalamic fibers had distinct and stereotyped growth patterns that related closely to their position; within the striatum growth cones were small and elongated, rarely extending filopodia or side branches. Axons grew quickly, in straight trajectories, with minimal pauses or retractions. When they reached the ventral intermediate zone, axons slowed down, often coming to a complete stop for up to several hours, and their growth cones became larger and more complex. During pauses there were continuous extensions and retractions of filopodia and/or side branches. When advance resumed, it was often to a different direction. These results demonstrate consistent regional variations in growth patterns that identify an unexpected decision region for thalamic axons. They provide the basis for examining the roles of guidance cues in an accessible yet intact preparation of the thalamocortical pathway and allow for an evaluation of previously suggested pathfinding mechanisms. PMID- 10804208 TI - From plaque to pretzel: fold formation and acetylcholine receptor loss at the developing neuromuscular junction. AB - Although there has been progress in understanding the initial steps in the formation of synapses, less is known about their subsequent maturation (Sanes and Lichtman, 1999). Two alterations on the postsynaptic side of the mammalian neuromuscular junction occur during early postnatal life: acetylcholine receptors (AChRs) disappear from parts of the developing junction as all but one axonal inputs are removed, and the topography of the postsynaptic membrane becomes more complicated as gutters and folds are established. We have studied the maturation of the AChR distribution and postsynaptic topography simultaneously by imaging labeled AChRs at the mouse neuromuscular junction in a new way, using reflected light confocal microscopy. At birth postsynaptic receptors were localized in irregular patches within a spoon-shaped plaque. Beginning several days later, receptor regions within a single endplate were divided into differentiated and less organized compartments. Folds generally oriented orthogonal to the long axis of the muscle fiber were seen in developing gutters, although the orientation of the gutters seemed to be imposed by the branching pattern of the nerve. Eventually, superficial regions lacking AChR labeling were apparent in all junctions. In junctions denervated in the neonatal period both gutter formation and the disappearance of superficial receptors regions were prevented. We suggest that tension between growing muscle fibers and the relatively inelastic synaptic terminals that adhere to them causes the topographic features of the postsynaptic membrane. This view provides a mechanical explanation for gutters, folds, and the location of folds at sites of neurotransmitter release. PMID- 10804210 TI - Neurodegeneration in Lurcher mice occurs via multiple cell death pathways. AB - Lurcher (Lc) is a gain-of-function mutation in the delta2 glutamate receptor (GRID2) that results in the cell-autonomous death of cerebellar Purkinje cells in heterozygous lurcher (+/Lc) mice. This in turn triggers the massive loss of afferent granule cells during the first few postnatal weeks. Evidence suggests that the death of Purkinje cells as a direct consequence of GRID2(Lc) activation and the secondary death of granule cells because of target deprivation occur by apoptosis. We have used mice carrying null mutations of both the Bax and p53 genes to examine the roles of these genes in cell loss in lurcher animals. The absence of Bax delayed Purkinje cell death in response to the GRID2(Lc) mutation and permanently rescued the secondary death of granule cells. In contrast, the p53 deletion had no effect on either cell death pathway. Our results demonstrate that target deprivation induces a Bax-dependent, p53-independent cell death response in cerebellar granule cells in vivo. In contrast, Bax plays a minor role in GRID2(Lc)-mediated Purkinje cell death. PMID- 10804209 TI - Recycling of the cell adhesion molecule L1 in axonal growth cones. AB - The cell adhesion molecule (CAM) L1 plays crucial roles in axon growth in vitro and in the formation of major axonal tracts in vivo. It is generally thought that CAMs link extracellular immobile ligands with retrogradely moving actin filaments to transmit force that pulls the growth cone forward. However, relatively little is known about the fate of CAMs that have been translocated into the central (C) domain of the growth cone. We have shown previously that L1 is preferentially endocytosed at the C-domain. In the present study, we further analyze the subcellular distribution of endocytic organelles containing L1 at different time points and demonstrate that internalized L1 is transported into the peripheral (P)-domain of growth cones advancing via an L1-dependent mechanism. Internalized L1 is found in vesicles positioned along microtubules, and the centrifugal transport of these L1-containing vesicles is dependent on dynamic microtubules in the P-domain. Furthermore, we show that endocytosed L1 is reinserted into the plasma membrane at the leading edge of the P-domain. Monitoring recycled L1 reveals that it moves retrogradely on the cell surface into the C-domain. In contrast, the growth cone advancing independently of L1 internalizes and recycles L1 within the C-domain. For the growth cone to advance, the leading edge needs to establish strong adhesive interactions with the substrate while attachments at the rear are released. Recycling L1 from the C-domain to the leading edge provides an effective way to create asymmetric L1-mediated adhesion and therefore would be critical for L1-based growth cone motility. PMID- 10804211 TI - Isolation and in vitro differentiation of conditionally immortalized murine olfactory receptor neurons. AB - Two major challenges exist in our understanding of the olfactory system. One concerns the enormous combinatorial code underlying odorant discrimination by odorant receptors. The other relates to neurogenesis and neuronal development in the olfactory epithelium. To address these issues, continuous cell cultures containing olfactory receptor neurons (ORNs) were obtained from olfactory epithelia of H-2K(b)-tsA58 transgenic mice. ORNs were detected and characterized by immunocytochemistry, RT-PCR, and Western blot for the markers Galpha(olf), adenylyl cyclase III, the olfactory cyclic nucleotide-gated channel subunits, and olfactory marker protein. In culture, epidermal growth factor and nerve growth factor stimulated proliferation, and brain-derived neurotrophic factor and neurotrophin-3 induced cellular maturation. Clonal cell lines were isolated by fluorescence-activated cell sorting with anti-neural cell adhesion molecule antibodies, and of 144 single cells plated, 39 clones were expanded, propagated, and stored in liquid nitrogen. All attempts at recovery of clonal lines from frozen stocks have been successful. The most thoroughly characterized clone, 3NA12, expressed ORN markers and responded to stimulation by single odorants. Each odorant activated approximately 1% of cells in a clonal line, and this suggests that many different odorant receptors may be expressed by these clonal cells. Therefore, these cell lines and the method by which they have been obtained represent a significant advance in the generation of olfactory cell cultures and provide a system to investigate odorant coding and olfactory neurogenesis. PMID- 10804212 TI - Wild-type huntingtin protects from apoptosis upstream of caspase-3. AB - Expansion of a polyglutamine sequence in the N terminus of huntingtin is the gain of-function event that causes Huntington's disease. This mutation affects primarily the medium-size spiny neurons of the striatum. Huntingtin is expressed in many neuronal and non-neuronal cell types, implying a more general function for the wild-type protein. Here we report that wild-type huntingtin acts by protecting CNS cells from a variety of apoptotic stimuli, including serum withdrawal, death receptors, and pro-apoptotic Bcl-2 homologs. This protection may take place at the level of caspase-9 activation. The full-length protein also modulates the toxicity of the poly-Q expansion. Cells expressing full-length mutant protein are susceptible to fewer death stimuli than cells expressing truncated mutant huntingtin. PMID- 10804213 TI - Neuronal basic helix-loop-helix proteins (NEX and BETA2/Neuro D) regulate terminal granule cell differentiation in the hippocampus. AB - The transcription factors neuronal helix-loop-helix protein (NEX)/mammalian atonal homolog 2 (Math-2), BETA2/neuronal determination factor (NeuroD), and NeuroD-related factor (NDRF)/NeuroD2 comprise a family of Drosophila atonal related basic helix-loop-helix (bHLH) proteins with highly overlapping expression in the developing forebrain. The ability of BETA2/NeuroD and NDRF to convert ectodermal cells into neurons after mRNA injection into Xenopus oocytes suggested a role in specifying neuronal cell fate. However, neuronal bHLH genes are largely transcribed in CNS neurons, which are fully committed. Here we analyze a defect in mice lacking BETA2/NeuroD, and in NEX*BETA2/NeuroD double mutants, demonstrating that bHLH proteins are required in vivo for terminal neuronal differentiation. Most strikingly, presumptive granule cells of the dentate gyrus are generated but fail to mature, lack normal sodium currents, and show little dendritic arborization. Long-term hippocampal slice cultures demonstrate secondary alterations of entorhinal and commissural/associational projections. The primary developmental arrest appears to be restricted to granule cells in which an autoregulatory system involving all three neuronal bHLH genes has failed. PMID- 10804214 TI - Cell contact regulates fate choice by cortical stem cells. AB - Cell fate is determined by intrinsic programs and external cues, such as soluble signals and cell-cell contact. Previous studies have demonstrated the roles of soluble factors in the proliferation and differentiation of cortical stem cells and cell-cell contact in maintaining stem cells in a proliferative state. In the present study, we focused on the effect of cell-cell interaction on cell-fate determination. We found that density could exert a strong influence on the cell type composition when cortical stem cells differentiate. Multipotent stem cells, which normally gave rise to neurons, astrocytes, and oligodendrocytes under high density culture condition, differentiated almost exclusively into smooth muscle at low density. Clonal analysis indicated that smooth muscle and astrocytes were derived from a common precursor and that the density effect on cell types used an instructive mechanism on the choice of fate rather than an effect of selective survival and/or proliferation. This instructive mechanism depended on the local and not the average density of the cells. This local signal could be mimicked by membrane extract. These findings demonstrate the importance of membrane-bound signals in specifying lineage and provide the first evidence for a short-range regulatory mechanism in cortical stem cell differentiation. PMID- 10804215 TI - Synapsin III: developmental expression, subcellular localization, and role in axon formation. AB - We have investigated the developmental expression and subcellular localization of synapsin III, the newest member of the synapsin family, in cultured mouse hippocampal neurons. Our results indicate that synapsin III is expressed early during development, with levels peaking 7 d after plating and declining thereafter. Synapsin III is highly concentrated in growth cones. Using specific antisense oligonucleotides, we have also examined the effect of depleting synapsin III on neurite elongation and synaptogenesis. When synapsin III was suppressed immediately after plating, hippocampal neurons extended minor processes but failed to differentiate one of them as the axon. The suppression of synapsin III after axonal elongation did not affect the time course of synapse formation. The results indicate that synapsin III has a developmental time course, a subcellular localization, and a developmental function very different from those of synapsin I and synapsin II. PMID- 10804216 TI - Olivocerebellar climbing fibers in the granuloprival cerebellum: morphological study of individual axonal projections in the X-irradiated rat. AB - Elimination of cerebellar granule cells early during postnatal development produces abnormal neural organization that retains immature characteristics in the adult, including innervation of each Purkinje cell by multiple climbing fibers from the inferior olive. To elucidate mechanisms underlying development of the olivocerebellar projection, we studied light-microscopic morphology of single olivocerebellar axons labeled with biotinylated dextran amine in adult rats rendered agranular by a single postnatal X-irradiation. Each reconstructed olivocerebellar axon gave off approximately 12 climbing fibers, approximately twice as many as in normal rats. Terminal arborizations of climbing fibers made irregular tufts in most areas, whereas they were arranged vertically in a few mildly affected areas. Each climbing fiber terminal arborization innervated only part of the dendritic arbor of a Purkinje cell, and multiple climbing fibers innervated a single Purkinje cell. These climbing fibers originated either from the same olivocerebellar axon (pseudomultiple innervation) or from distinct axons (true multiple innervation). Abundant non-climbing fiber thin collaterals projected to all cortical layers. Although the longitudinal pattern of the zonal olivocerebellar projection was generally observed, lateral branching, including bilateral projections, was relatively frequent. These results suggest that the granule cell-parallel fiber system induces several important features of olivocerebellar projection: (1) organization of the climbing fiber terminal arborization tightly surrounding Purkinje cell dendrites, (2) elimination of pseudo- and true multiple innervations establishing one-to-one innervation, (3) retraction of non-climbing fiber thin collaterals from the molecular layer, and (4) probable refinement of the longitudinal projection domains by removing aberrant transverse branches. PMID- 10804217 TI - Encoding of tactile stimulus location by somatosensory thalamocortical ensembles. AB - The exquisite modular anatomy of the rat somatosensory system makes it an excellent model to test the potential coding strategies used to discriminate the location of a tactile stimulus. Here, we investigated how ensembles of simultaneously recorded single neurons in layer V of primary somatosensory (SI) cortex and in the ventral posterior medial (VPM) nucleus of the thalamus of the anesthetized rat may encode the location of a single whisker stimulus on a single trial basis. An artificial neural network based on a learning vector quantization algorithm, was used to identify putative coding mechanisms. Our data suggest that these neural ensembles may rely on a distributed coding scheme to represent the location of single whisker stimuli. Within this scheme, the temporal modulation of neural ensemble firing rate, as well as the temporal interactions between neurons, contributed significantly to the representation of stimulus location. The relative contribution of these temporal codes increased with the number of whiskers that the ensembles must discriminate among. Our results also indicated that the SI cortex and the VPM nucleus may function as a single entity to encode stimulus location. Overall, our data suggest that the representation of somatosensory features in the rat trigeminal system may arise from the interactions of neurons within and between the SI cortex and VPM nucleus. Furthermore, multiple coding strategies may be used simultaneously to represent the location of tactile stimuli. PMID- 10804218 TI - Association of beta 1 integrin with focal adhesion kinase and paxillin in differentiating Schwann cells. AB - Schwann cells (SCs) differentiate into a myelinating cell when simultaneously adhering to an axon destined for myelination and basal lamina. We are interested in defining the signaling pathway activated by basal lamina. Using SC/sensory neuron (N) cocultures, we identified beta1 integrin and F-actin as components of a pathway leading to myelin gene expression and myelination (Fernandez-Valle et al., 1994, 1997). Here, we show that focal adhesion kinase (FAK) and paxillin are constitutively expressed by SCs contacting axons in the absence of basal lamina. Tyrosine phosphorylation of FAK and paxillin increases as SCs form basal lamina and differentiate. FAK and paxillin specifically coimmunoprecipitate with beta1 integrin in differentiating SC/N cocultures but not SC-only cultures. Paxillin coimmunoprecipitates with FAK and fyn kinase in differentiating SC/N cocultures. A subset of tyrosine-phosphorylated beta1 integrin, FAK, and paxillin molecules reside in the insoluble, F-actin-rich fraction of differentiating cocultures. Cytochalasin D, an actin depolymerizing agent, decreases tyrosine phosphorylation of FAK and paxillin and their association with beta1 integrin and causes a dose dependent increase in the abundance of insoluble FAK and paxillin complexes. Collectively, our work indicates that beta1 integrin, FAK, paxillin, and fyn kinase form an actin-associated complex in SCs adhering to basal lamina in the presence of axons. This complex may be important for initiating the process of SC differentiation into a myelinating cell. PMID- 10804219 TI - Distribution, targeting, and internalization of the sst4 somatostatin receptor in rat brain. AB - Somatostatin mediates its diverse physiological effects through a family of five G-protein-coupled receptors (sst(1)-sst(5)); however, knowledge about the distribution of individual somatostatin receptor proteins in mammalian brain is incomplete. In the present study, we have examined the regional and subcellular distribution of the somatostatin receptor sst(4) in the rat CNS by raising anti peptide antisera to the C-terminal tail of sst(4). The specificity of affinity purified antibodies was demonstrated using immunofluorescent staining of HEK 293 cells stably transfected with an epitope-tagged sst(4) receptor. In Western blotting, the antiserum reacted specifically with a broad band in rat brain, which migrated at approximately 70 kDa before and approximately 50 kDa after enzymatic deglycosylation. sst(4)-Like immunoreactivity was most prominent in many forebrain regions, including the cerebral cortex, hippocampus, striatum, amygdala, and hypothalamus. Analysis at the electron microscopic level revealed that sst(4)-expressing neurons target this receptor preferentially to their somatodendritic domain. Like the sst(2A) receptor, sst(4)-immunoreactive dendrites were often closely apposed by somatostatin-14-containing fibers and terminals. However, unlike the sst(2A) receptor, sst(4) was not internalized in response to intracerebroventricular administration of somatostatin-14. After percussion trauma of the cortex, neuronal sst(4) receptors progressively declined at the sites of damage. This decline coincided with an induction of sst(4) expression in cells with a glial-like morphology. Together, this study provides the first description of the distribution of immunoreactive sst(4) receptor proteins in rat brain. We show that sst(4) is strictly somatodendritic and most likely functions in a postsynaptic manner. In addition, the sst(4) receptor may have a previously unappreciated function during the neuronal degeneration regeneration process. PMID- 10804221 TI - Wind direction coding in the cockroach escape response: winner does not take all. AB - Cockroaches respond to the approach of a predator by turning away and then running. Three bilateral pairs of giant interneurons are involved in determining the direction of the sensory stimulus and setting the turn direction. Each of these six interneurons has a different directional response to wind stimuli. We have tested whether these six cells use a winner-take-all mechanism to perform this directional determination: that is, each of these cells suppressing the motor response that each of the other cells promotes. Such a mechanism is found in similar behaviors of some other animals. By adding spikes to identified giant interneurons through intracellular stimulation during the sensory-induced behavior and analyzing the resulting directional leg movements, we find that a winner-take-all is not used in this system. Rather, directional determination appears to be based on collaborative calculation of direction by the giant interneurons as a group. PMID- 10804220 TI - Convergent inputs from thalamic motor nuclei and frontal cortical areas to the dorsal striatum in the primate. AB - Current models of basal ganglia circuitry primarily associate the ventral thalamic nuclei with relaying basal ganglia output to the frontal cortex. However, some studies have demonstrated projections from the ventral anterior (VA) and ventral lateral (VL) thalamic nuclei to the striatum, suggesting that these nuclei directly modulate the striatum. VA/VL nuclei have specific connections with primary, supplementary, premotor, and cingulate motor cortices indicating their involvement in motor function. These areas mediate different aspects of motor control such as movement execution, motor learning, and sensorimotor integration. Increasing evidence indicates that functionally related motor areas have convergent projections to the dorsal striatum, suggesting that integration of different aspects of motor control occur at the level of the striatum. This study examines the organization of VA/VL thalamic inputs to the dorsal "motor" striatum to determine how this afferent projection is organized with respect to corticostriatal afferents from motor, premotor, and cingulate motor areas. Motor cortical projections to specific dorsal striatal regions arose from multiple areas, including components from primary motor, premotor, supplementary, and cingulate motor areas. Diverse motor cortical projections to a given dorsal striatal region indicated convergence of functionally related corticostriatal motor pathways. Most dorsal striatal sites received dense thalamic inputs from the VL pars oralis nucleus. Additional thalamostriatal projections arose from VA, VL pars caudalis, and ventral posterior lateral pars oralis nuclei and Olszewski's Area X. Our results provide evidence for convergent striatal projections from interconnected ventral thalamic and cortical motor areas, suggesting that these afferents modulate the same striatal output circuits. PMID- 10804222 TI - Population vector coding by the giant interneurons of the cockroach. AB - We tested two alternative models of integration among the cockroach giant interneurons (GIs) for determining the directions of wind-evoked escape turns. One model, called steering wheel, pits contralateral GIs against one another; the other, called population vector model, involves a vector computation among the GIs. In testing each model theoretically, the population vector was found to account far better for the actual behavior. Both models could account for the results of previous behavioral-physiological experiments in which spikes had been added to the right GI3 together with wind stimuli from the right side. The two models revealed a critical behavioral-physiological experimental test that we then performed; namely, when delivering wind from the right side, adding spikes experimentally to the right GI2 should increase turn size according to the steering wheel model but should decrease turn size according to the population vector model. The latter result was obtained. The population vector, but not the steering wheel, model also could account for previous behavioral-physiological experiments in which spikes were added experimentally to a GI contralateral to the wind stimuli. The results support the population vector model as accounting for direction determination among the cockroach GIs. PMID- 10804223 TI - Effect of lesions of the ventrolateral preoptic nucleus on NREM and REM sleep. AB - Neurons in the ventrolateral preoptic nucleus (VLPO) in rats show c-fos activation after sleep and provide GABAergic innervation of the major monoamine arousal systems, suggesting that they may be a necessary part of the brain circuitry that produces sleep. We examined the effects on sleep behavior in rats of cell-specific damage to the VLPO by microinjection of ibotenic acid. Severe lesions of the central cell cluster of the VLPO ( approximately 80-90% cell loss bilaterally) caused a 60-70% decrease in delta power and a 50-60% decrease in nonrapid-eye-movement (NREM) sleep time (p < 0.001). The number of remaining Fos immunoreactive neurons in the VLPO cell cluster was linearly related to NREM sleep time (r = 0.77; p < 0.001) and total electroencephalogram delta power (r = 0. 79; p < 0.001) but not to rapid-eye-movement (REM) sleep (r = 0.35; p > 0.10). Lesions in the region containing scattered VLPO neurons medial or dorsal to the cell cluster caused smaller changes in NREM sleep time (24.5 or 15%, respectively) but were more closely associated with loss of REM sleep (r = 0.74; p < 0.01). The insomnia caused by bilateral VLPO lesions persisted for at least 3 weeks. Lesions of the VLPO caused no change in mean body temperature or its circadian variation; after small lesions of the ventromedial preoptic nucleus, body temperature showed normal circadian variation but a wider temperature range, and sleep behavior was not affected. These experiments delineate distinct preoptic sites with primary effects on the regulation of NREM sleep, REM sleep, and body temperature. PMID- 10804224 TI - Glucocorticoid negative feedback selectively targets vasopressin transcription in parvocellular neurosecretory neurons. AB - To identify molecular targets of corticosteroid negative feedback effects on neurosecretory neurons comprising the central limb of the hypothalamo-pituitary adrenal (HPA) axis, we monitored ether stress effects on corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) heteronuclear RNA (hnRNA) expression in rats that were intact or adrenalectomized (ADX) and replaced with corticosterone (B) at constant levels ranging from nil to peak stress concentrations. Under basal conditions, relative levels of both primary transcripts varied inversely as a function of plasma B titers. In response to stress, the kinetics of CRF hnRNA responses of intact and ADX rats replaced with low B were similar, peaking at 5 min after stress. By contrast, intact rats showed a delayed AVP hnRNA response (peak at 2 hr), the timing of which was markedly advanced in ADX/low B-replaced animals (peak at 5-30 min). Transcription factors implicated in these responses responded similarly. Manipulation of B status did not affect the early (5-15 min) phosphorylation of transcription factor cAMP-response element-binding protein (CREB) but accelerated maximal Fos induction from 2 hr after stress (intact) to 1 hr (ADX). Assays of binding by proteins in hypothalamic extracts of similarly manipulated rats toward consensus CRE and AP-1 response elements supported a role for the stress-induced plasma B increment in antagonizing AP-1, but not CRE, binding. These findings suggest that glucocorticoid negative feedback at the transcriptional levels is exerted selectively on AVP gene expression through a mechanism that likely involves glucocorticoid receptor interactions with immediate-early gene products. PMID- 10804225 TI - Contrasting effects on discrimination learning after hippocampal lesions and conjoint hippocampal-caudate lesions in monkeys. AB - Eighteen monkeys with lesions of the hippocampal region (the hippocampus proper, the dentate gyrus, and the subiculum) made by an ischemic procedure, radio frequency, or ibotenic acid were tested on a simple, two-choice object discrimination learning task that has been shown to be sensitive to large lesions of the medial temporal lobe. The monkeys were also tested on two other discrimination tasks (pattern discrimination and eight-pair concurrent discrimination) that can be learned normally by monkeys with large medial temporal lobe lesions. All of the lesion groups were impaired at learning the simple object discrimination task. Seven of the monkeys who had sustained damage to the hippocampal region also sustained damage to the tail of the caudate nucleus. These seven monkeys, but not the other 11 monkeys with hippocampal lesions, were impaired on pattern discrimination and concurrent discrimination learning. The results suggest that the hippocampal region is important for learning easy, two-choice discriminations, whereas the caudate nucleus is necessary for the normal learning of more difficult, gradually acquired discrimination tasks. The findings support the distinction between declarative memory, which depends on the hippocampus and related medial temporal lobe structures, and habit learning, which depends on the caudate nucleus. PMID- 10804226 TI - Projections from the rat prefrontal cortex to the ventral tegmental area: target specificity in the synaptic associations with mesoaccumbens and mesocortical neurons. AB - Excitatory projections from the prefrontal cortex (PFC) to the ventral tegmental area (VTA) play an important role in regulating the activity of VTA neurons and the extracellular levels of dopamine (DA) within forebrain regions. Previous investigations have demonstrated that PFC terminals synapse on the dendrites of DA and non-DA neurons in the VTA. However, the projection targets of these cells are not known. To address whether PFC afferents innervate different populations of VTA neurons that project to the nucleus accumbens (NAc) or to the PFC, a triple labeling method was used that combined peroxidase markers for anterograde and retrograde tract-tracing with pre-embedding immunogold-silver labeling for either tyrosine hydroxylase (TH) or GABA. Within the VTA, PFC terminals formed asymmetric synapses onto dendritic shafts that were immunoreactive for either TH or GABA. PFC terminals also synapsed on VTA dendrites that were retrogradely labeled from the NAc or the PFC. Dendrites retrogradely labeled from the NAc and postsynaptic to PFC afferents were sometimes immunoreactive for GABA but were never TH-labeled. Conversely, dendrites retrogradely labeled from the PFC and postsynaptic to PFC afferents were sometimes immunoreactive for TH but were never GABA-labeled. These results provide the first demonstration of PFC afferents synapsing on identified cell populations in the VTA and indicate a considerable degree of specificity in the targets of the PFC projection. The unexpected finding of selective PFC synaptic input to GABA-containing mesoaccumbens neurons and DA-containing mesocortical neurons suggests novel mechanisms through which the PFC can influence the activity of ascending DA and GABA projections. PMID- 10804227 TI - Impact of self-administered cocaine and cocaine cues on extracellular dopamine in mesolimbic and sensorimotor striatum in rhesus monkeys. AB - Studies were conducted to determine the impact of self-administered cocaine on extracellular striatal dopamine in four rhesus monkeys. The extent to which external cue conditioning contributed to the effects of cocaine and whether there is activation of striatal dopaminergic neurotransmission during drug-seeking behavior was also examined. Microdialysis measurements were made at 2 min intervals in sensorimotor (dorsolateral) and mesolimbic (central and ventromedial) striatum. A fixed-ratio schedule of reinforcement was used, with cocaine availability signaled by a visual cue. Studies examined the effects of cocaine or cocaine cues against a drug-free baseline. Large (fivefold to eightfold) increases in extracellular dopamine after a self-administered infusion of 0.5 mg/kg cocaine were quite rapid and matched the time course of reported subjective effects in human laboratory studies. To determine if conditioning to external cues contributed to the cocaine-induced increases, saline was substituted for cocaine in the infusion, leaving all other visual and auditory stimuli unchanged. No increase in extracellular dopamine in either sensorimotor or mesolimbic striatal subdivisions was observed. Extracellular dopamine during extended periods of drug-seeking behavior triggered by a visual cue was determined in both central and ventromedial striatum. This procedure also did not result in any measurable changes in extracellular dopamine. These studies demonstrate rapid and pronounced pharmacological actions of self-administered cocaine. No apparent conditioned component of those actions was associated with external environmental cues, suggesting that cues that trigger drug-seeking behavior in nonhuman primates do not cause conditioned increases in mesolimbic striatal dopamine. PMID- 10804228 TI - Progressive transneuronal changes in the brainstem and thalamus after long-term dorsal rhizotomies in adult macaque monkeys. AB - This study deals with a potential brainstem and thalamic substrate for the extensive reorganization of somatosensory cortical maps that occurs after chronic, large-scale loss of peripheral input. Transneuronal atrophy occurred in neurons of the dorsal column (DCN) and ventral posterior lateral thalamic (VPL) nuclei in monkeys subjected to cervical and upper thoracic dorsal rhizotomies for 13-21 years and that had shown extensive representational plasticity in somatosensory cortex and thalamus in other experiments. Volumes of DCN and VPL, number and sizes of neurons, and neuronal packing density were measured by unbiased stereological techniques. When compared with the opposite, unaffected, side, the ipsilateral cuneate nucleus (CN), external cuneate nucleus (ECN), and contralateral VPL showed reductions in volume: 44-51% in CN, 37-48% in ECN, and 32-38% in VPL. In the affected nuclei, neurons were progressively shrunken with increasing survival time, and their packing density increased, but there was relatively little loss of neurons (10-16%). There was evidence for loss of axons of atrophic CN cells in the medial lemniscus and in the thalamus, with accompanying severe disorganization of the parts of the ventral posterior nuclei representing the normally innervated face and the deafferented upper limb. Secondary transneuronal atrophy in VPL, associated with retraction of axons of CN neurons undergoing primary transneuronal atrophy, is likely to be associated with similar withdrawal of axons from the cerebral cortex and should be a powerful influence on reorganization of somatotopic maps in the somatosensory cortex. PMID- 10804229 TI - Cholinergic excitation of septohippocampal GABA but not cholinergic neurons: implications for learning and memory. AB - The medial septum/diagonal band (MSDB), which gives rise to the septohippocampal pathway, is a critical locus for the mnemonic effects of muscarinic drugs. Infusion of muscarinic cholinergic agonists into the MSDB enhance learning and memory processes both in young and aged rats and produce a continuous theta rhythm in the hippocampus. Intraseptal muscarinic agonists also alleviate the amnesic syndrome produced by systemic administration of muscarinic receptor antagonists. It has been presumed, but not proven, that the cellular mechanisms underlying the effects of muscarinic agonists in the MSDB involve an excitation of septohippocampal cholinergic neurons and a subsequent increase in acetylcholine (ACh) release in the hippocampus. Using a novel fluorescent labeling technique to selectively visualize live septohippocampal cholinergic neurons in rat brain slices, we have found that muscarinic agonists do not excite septohippocampal cholinergic neurons, instead they inhibit a subpopulation of cholinergic neurons. In contrast, unlabeled neurons, confirmed to be noncholinergic, septohippocampal GABA-type neurons using retrograde marking and double-labeling techniques, are profoundly excited by muscarine. Thus, the cognition-enhancing effects of muscarinic drugs in the MSDB cannot be attributed to an increase in hippocampal ACh release. Instead, disinhibitory mechanisms, caused by increased impulse flow in the septohippocampal GABAergic pathway, may underlie the cognition-enhancing effects of muscarinic agonists. PMID- 10804230 TI - Dendritic depolarization efficiently attenuates low-threshold calcium spikes in thalamic relay cells. AB - Thalamic relay cells respond in two distinct modes, burst and tonic, that depend on a voltage-dependent, low-threshold, transient Ca(2+) current (I(T)), and these modes relay different forms of information to cortex. I(T) activation evokes a low-threshold spike (LTS), producing a burst of action potentials. Modulatory inputs from cortex and brainstem are known to activate metabotropic receptors on relay cell dendrites at which the T channels underlying I(T) may be concentrated. We thus investigated the influence of activating these receptors on the LTS, using current-clamp intracellular recording in an in vitro slice preparation of the cat's lateral geniculate nucleus. We found a strong correlation between LTS amplitude and the number of action potentials evoked in the burst. We then found that activation of either metabotropic glutamate or muscarinic receptors produced a hyperpolarizing shift in the sigmoid relationship between LTS amplitude and the initial holding potential without affecting the maximum LTS amplitude or slope of the relationship. This hyperpolarizing shift in the voltage dependency of LTS amplitude is best explained by space-clamp limitations and significantly more depolarization of T channels near the dendritic location of activated receptors than at the soma. Thus, nonretinal modulatory inputs may have a stronger influence on I(T) and number of action potentials generated in a burst than previously imagined from somatic recording, because the EPSP amplitudes generated by these inputs at the dendritic location of most T channels are greater than after their electrotonic decay recorded at the soma. PMID- 10804232 TI - Postnatal handling increases the expression of cAMP-inducible transcription factors in the rat hippocampus: the effects of thyroid hormones and serotonin. AB - Postnatal handling increases glucocorticoid receptor expression in the rat hippocampus, thus altering the regulation of hypothalamic synthesis of corticotropin-releasing hormone and the hypothalamic-pituitary-adrenal response to stress. The effect on glucocorticoid receptor gene expression represents one mechanism by which the early environment can exert a long-term effect on neural development. The handling effect on hippocampal glucocorticoid receptor expression is dependent on peripheral thyroid hormone release and the activation of ascending serotonergic pathways. In primary hippocampal cell cultures, serotonin (5-HT) increases glucocorticoid receptor expression, and this effect appears to be mediated by increased cAMP levels. In the current studies we examined the in vivo effects of handling on hippocampal cAMP-protein kinase A (PKA) activity. In 7-d-old rat pups, we found that (1) postnatal handling increased adenylyl cyclase activity and hippocampal cAMP levels, (2) the effect of handling on cAMP levels was completely blocked by treatment with either propylthiouracil (PTU), a thyroid hormone synthesis inhibitor, or the 5-HT receptor antagonist, ketanserin, and (3) handling also increased hippocampal PKA activity. We then examined the effects of handling on cAMP-inducible transcription factors. Handling rapidly increased levels of the mRNAs for nerve growth factor-inducible factor A (NGFI-A) (zif268, krox24) and activator protein 2 (AP-2) as well as for NGFI-A and AP-2 immunoreactivity throughout the hippocampus. Finally, we found that the effects of handling on NGFI-A and AP-2 expression were significantly reduced by concurrent treatment with either PTU or ketanserin, effects that paralleled those on cAMP formation. NGFI-A and AP-2 have been implicated in the regulation of glucocorticoid receptor expression during development. Thus, these findings suggest that postnatal handling might alter glucocorticoid receptor gene expression via cAMP-PKA pathways involving the activation of NGFI-A and AP-2. PMID- 10804233 TI - Application of Live Monocells from Macroalgae to Shellfish Seed Production. AB - Monocells were isolated from several macroalgae, Porphyra yezoensis, Undaria pinnatifida, and Laminaria japonica, by digestion with alga-tool enzymes. The monocells were then used to feed the parents or larvae of bay scallop Argopecten irradians, blood cockle Arca inflata, and abalone Haliotis discus juveniles. Results showed that the parents of bay scallop and blood cockle fed with Porphyra monocells could mature and discharge eggs and spermatozoa and their larvae could metamorphose; the survival rate of abalone juveniles fed with isolated cells from Laminaria and Undaria increased by 100% compared with that of those fed with artificial food. PMID- 10804231 TI - Do glia have heart? Expression and functional role for ether-a-go-go currents in hippocampal astrocytes. AB - Potassium homeostasis plays an important role in the control of neuronal excitability, and diminished buffering of extracellular K results in neuronal Hyperexcitability and abnormal synchronization. Astrocytes are the cellular elements primarily involved in this process. Potassium uptake into astrocytes occurs, at least in part, through voltage-dependent channels, but the exact mechanisms involved are not fully understood. Although most glial recordings reveal expression of inward rectifier currents (K(IR)), it is not clear how spatial buffering consisting of accumulation and release of potassium may be mediated by exclusively inward potassium fluxes. We hypothesized that a combination of inward and outward rectifiers cooperate in the process of spatial buffering. Given the pharmacological properties of potassium homeostasis (sensitivity to Cs(+)), members of the ether-a-go-go (ERG) channel family widely expressed in the nervous system could underlie part of the process. We used electrophysiological recordings and pharmacological manipulations to demonstrate the expression of ERG-type currents in cultured and in situ hippocampal astrocytes. Specific ERG blockers (dofetilide and E 4031) inhibited hyperpolarization- and depolarization-activated glial currents, and ERG blockade impaired clearance of extracellular potassium with little direct effect on hippocampal neuron excitability. Immunocytochemical analysis revealed ERG protein mostly confined to astrocytes; ERG immunoreactivity was absent in presynaptic and postsynaptic elements, but pronounced in glia surrounding the synaptic cleft. Oligodendroglia did not reveal ERG immunoreactivity. Intense immunoreactivity was also found in perivascular astrocytic end feet at the blood-brain barrier. cDNA amplification showed that cortical astrocytes selectively express HERG1, but not HERG2-3 genes. This study provides insight into a possible physiological role of hippocampal ERG channels and links activation of ERG to control of potassium homeostasis. PMID- 10804234 TI - Antioxidant Peptides from the Protease Digest of Prawn (Penaeus japonicus) Muscle. AB - The muscle of the prawn Penaeus japonicus was hydrolyzed by various proteases, and antioxidant activity of the hydrolysates was examined. Among the digests, pepsin digest showed the most potent antioxidant activity. Three antioxidant peptides have been isolated from the active peptidic fraction by ion-exchange chromatography, gel filtration, and ODS high-performance liquid chromatography. Their structures were identified as Ile-Lys-Lys, Phe-Lys-Lys, and Phe-Ile-Lys Lys. PMID- 10804235 TI - Development of an Argopecten-Specific 18S rRNA Targeted Genetic Probe. AB - Comparison of 18S ribosomal RNA gene sequences between diverse bivalve species, including eight scallop species, allowed the design of an 18S rRNA targeted oligonucleotide probe (BS-1364) that was specific for scallops belonging to the genus Argopecten (bay and calico scallops). The high sequence similarity of the 18S rRNA gene between Argopecten irradians and Argopecten gibbus (98.8%) prevented the design of an A. irradians species-specific probe. Hybridization studies using amplified 18S rDNA from a diverse collection of bivalve species demonstrated that the specificity of the digoxygenin-labeled probe was consistent with the predicted specificity indicated by sequence comparison. Hybridization studies using laboratory-spawned bay scallop veligers indicated that a single veliger could be detected by probe hybridization in a blot format, and that probe hybridization signal was proportional (r(2) =.99) to the abundance of veligers. Methods for rRNA extraction and blotting were developed that allowed bay scallop veligers to be specifically and quantitatively identified in natural plankton samples. Preliminary studies conducted in Tampa Bay, Florida, suggest that introduced scallops can successfully spawn and produce veligers under in situ conditions. The Argopecten-specific probe and methods developed in this study provide the means to study the production and fate of bay scallop larvae in nature and provide evidence that scallops introduced into Tampa Bay have the potential for successful reproduction and enhancement of scallop stocks. PMID- 10804236 TI - Molecular Cloning of Growth Hormone Complementary DNA in Barfin Flounder (Verasper moseri). AB - The olive flounder (family Paralichthidae; Paralichthys olivaceus) growth hormone (ofGH) appears to be the most derived among known growth hormones, with the deletion of 14 consecutive amino acids in the carboxy-terminal region. To ascertain if this deletion is common to all flounders, growth hormone complementary DNA of the barfin flounder (bfGH) (family Pleuronectidae; Verasper moseri) has been cloned. It was amplified by polymerase chain reaction using single-strand cDNA from the pituitary gland. Excluding the poly(A) tail, the bfGH cDNA is 919 nucleotides long and contains a 609-bp open reading frame encoding a putative signal peptide of 17 amino acids and a mature protein of 186 amino acids. Northern blot analysis detected 1.0 kb of bfGH messenger RNA in the pituitary gland, which is a reasonable value considering the poly(A) tail. The deduced amino acid sequence of bfGH has 78% identity with the sequence of ofGH. A major difference is the presence of a 14 amino acid segment (140-153) in bfGH, as in other growth hormones, suggesting that this deletion in the olive flounder occurred after the divergence of the Pleuronectoidae. PMID- 10804237 TI - Cell Culture Evaluation of the Semliki Forest Virus Expression System As a Novel Approach for Antigen Delivery and Expression in Fish. AB - Heterologous gene expression by Semliki Forest virus (SFV) expression vectors was investigated in fish cell culture. Experiments performed using an infectious strain of SFV, replication-defective SFV particles, and recombinant SFV RNA constructs encoding the Escherchia coli LacZ or firefly luciferase reporter genes indicated that levels of SFV-mediated expression in fish cells were dependent on cell type and temperature. Maximal expression levels were observed in the two salmonid-derived cell lines CHSE-214 and F95/9 at 25 degrees C and 20 degrees C. As the temperature was lowered to 15 degrees C or below, levels of reporter gene expression were reduced up to 1000-fold, indicating that the SFV replication complex functioned inefficiently at low temperatures. The ability of SFV expression systems to function in fish cells was further investigated by analyzing the expression of the protective VP2 antigen of infectious pancreatic necrosis virus (IPNV) from the various constructs, including a novel DNA-based SFV plasmid. The VP2 protein produced in CHSE-214 and F95/9 cells transfected or infected with the recombinant SFV-IPNV VP2 constructs appeared to be synthesized in an antigenically correct form, as evidenced by the ability to react with several conformation-dependent IPNV-specific monoclonal antibodies. Whether the temperature-restricted replication and expression displayed by SFV-based constructs in fish cell culture also occurs in vivo remains to be determined. PMID- 10804238 TI - Characterization of Satellite DNA Sequences from the Commercially Important Marine Rotifers Brachionus rotundiformis and Brachionus plicatilis. AB - This study was initiated to search for species-specific and strain-specific satellite DNA sequences for which oligonucleotide primers could be designed to differentiate between various commercially important strains of the marine monogonont rotifers Brachionus rotundiformis and Brachionus plicatilis. Two unrelated, highly reiterated satellite sequences were cloned and characterized. The eight sequenced monomers from B. rotundiformis and six from B. plicatilis had low intrarepeat variability and were similar in their overall lengths, A + T compositions, and high degrees of repeated motif substructure. However, hybridizations to 19 representative strains, sequence characterizations, and GenBank searches indicated that these two satellites are morphotype-specific and population-specific, respectively, and share little homology to each other or to other characterized sequences in the database. Primer pairs designed for the B. rotundiformis satellite confirmed hybridization specificities on polymerase chain reaction and could serve as a useful molecular diagnostic tool to identify strains belonging to the SS morphotype, which are gaining widespread usage as first feeds for marine fish in commercial production. PMID- 10804239 TI - Identification of Microsatellite Repeats in Turbot (Scophthalmus maximus) and Dover Sole (Solea solea) using a RAPD-Based Technique: Characterization of Microsatellite Markers in Dover Sole. AB - We have used a RAPD-based technique to identify several microsatellite repeats in turbot (Scophthalmus maximus) and Dover sole (Solea solea) and report the characterization of six novel polymorphic microsatellite markers for Dover sole. These are the first such markers to be developed for this flatfish species. They exhibit much higher levels of heterozygosity than those previously observed with allozyme loci and should prove useful in addressing population genetic questions as well as more fundamental aquaculture-related questions. PMID- 10804240 TI - Purification and Characterization of N-Acetylglucosamine-6-phosphate Deacetylase from a Psychrotrophic Marine Bacterium, Alteromonas Species. AB - A psychrotrophic bacterium, strain Mct-9, which produced an N-acetylglucosamine-6 phosphate deacetylase, was isolated from a deep-seawater sample in the Mariana Trough. The Mct-9 strain was identified as Alteromonas sp. The native enzyme had a molecular mass of 164,000 Da, and was predicted to be composed of four identical subunits with molecular masses of 41,000 Da. The purified enzyme hydrolyzed N-acetylglucosamine (GlcNAc), GlcNAc-6-phosphate, and GlcNAc-6 sulfate. Considering the low K(m) and high k(cat)/K(m) for GlcNAc-6-phosphate, it probably acts as a GlcNAc-6-phosphate deacetylase in vivo. The enzyme was functional in the temperature range of 5 degrees to 70 degrees C and displayed optimal activity at 55 degrees C. The optimal temperature was higher than that of the deacetylase from the mesophilic bacterium Vibrio cholerae non-O1. The characteristics of the GlcNAc-6-phosphate deacetylase from Alteromonas sp. are unique among psychrotrophs and psychrophiles, whose intracellular enzymes are mostly thermolabile. PMID- 10804241 TI - Cloning and Sequence Analysis of Vibrio halioticoli Genes Encoding Three Types of Polyguluronate Lyase. AB - The alginate lyase-coding genes of Vibrio halioticoli IAM 14596(T), which was isolated from the gut of the abalone Haliotis discus hannai, were cloned using plasmid vector pUC 18, and expressed in Escherichia coli. Three alginate lyase positive clones, pVHB, pVHC, and pVHE, were obtained, and all clones expressed the enzyme activity specific for polyguluronate. Three genes, alyVG1, alyVG2, and alyVG3, encoding polyguluronate lyase were sequenced: alyVG1 from pVHB was composed of a 1056-bp open reading frame (ORF) encoding 352 amino acid residues; alyVG2 gene from pVHC was composed of a 993-bp ORF encoding 331 amino acid residues; and alyVG3 gene from pVHE was composed of a 705-bp ORF encoding 235 amino acid residues. Comparison of nucleotide and deduced amino acid sequences among AlyVG1, AlyVG2, and AlyVG3 revealed low homologies. The identity value between AlyVG1 and AlyVG2 was 18.7%, and that between AlyVG2 and AlyVG3 was 17.0%. A higher identity value (26.0%) was observed between AlyVG1 and AlyVG3. Sequence comparison among known polyguluronate lyases including AlyVG1, AlyVG2, and AlyVG3 also did not reveal an identical region in these sequences. However, AlyVG1 showed the highest identity value (36.2%) and the highest similarity (73.3%) to AlyA from Klebsiella pneumoniae. A consensus region comprising nine amino acid (YFKAGXYXQ) in the carboxy-terminal region previously reported by Mallisard and colleagues was observed only in AlyVG1 and AlyVG2. PMID- 10804242 TI - In Situ Polymerase Chain Reaction Visualization of Vibrio halioticoli Using Alginate Lyase Gene AlyVG2. AB - A prokaryotic in situ polymerase chain reaction (PI-PCR) technique was applied to visualize Vibrio halioticoli cells using alginate lyase gene alyVG2 as a target gene. Prior to PI-PCR, a primer set, VG2-OS3, for specific amplification of an approximately 1.0-kb fragment from V. halioticoli genomic DNA was developed with amplified fragments from V. pelagius and V. fischeri DNAs as reference strains. One-stage PI-PCR using the primer set, digoxigenin-labeled dUTP, and indirect alkaline-phosphatase-linked fluorescence detection technique (HNPP/Fast Red TR as a substrate) failed to differentiate V. halioticoli IAM14596(T) cells from ATCC25916(T) cells of the closely related species V. pelagius. However, two-stage PI-PCR adding the extension and digoxigenin-labeling step of the amplified fragment into the first amplification stage allowed us to differentiate V. halioticoli cells from V. pelagius cells. PMID- 10804243 TI - Five Crustacean Hyperglycemic Family Hormones of Penaeus monodon: Complementary DNA Sequence and Identification in Single Sinus Glands by Electrospray Ionization Fourier Transform Mass Spectrometry. AB - Five novel neuropeptides, designated Pm-sgp-I to -V, of the crustacean hyperglycemic hormone (CHH) family have been identified from the giant tiger prawn Penaeus monodon by isolation of the preprohormone genes from an eyestalk complementary DNA library. On the basis of sequence similarity, the encoded peptides have been classified as CHH-like type I hormones, which include all known CHHs and the molt-inhibiting hormone (MIH) of the lobster Homarus americanus. Consistent with CHH type I preprohormones, the Pm-sgp precursors include a signal peptide, a CHH precursor-related peptide (CPRP), and the CHH like hormone. Analysis by electrospray ionization-Fourier transform mass spectrometry enabled the neuropeptide complement of individual sinus glands to be resolved. It also confirmed the presence of the five Pm-sgp neuropeptides within the sinus gland of an individual animal, in that the masses observed were consistent with those predicted from the gene sequence of the Pm-sgps after posttranslational modification. These modifications included cleavage of the signal peptide and precursor protein, carboxy-terminal amidation, and formation of three disulfide bridges. Analysis of crude extracts of single sinus glands from different animals revealed variation in neuropeptide content and will provide a tool for determining whether the content varies as a function of the physiological state of the animal. PMID- 10804244 TI - Isolation and Characterization of Highly Polymorphic Microsatellites from the Polychaete Pectinaria koreni. AB - We report on the isolation of dinucleotide microsatellites from the polychaete Pectinaria koreni using (GT)(10) and (CT)(10) olignonucleotide probes. Compound and particularly imperfect microsatellites are predominant in this species. In most cases the associated element is (AT)(n) leading to AT microsatellites being the most frequent class after GT repeats. Obtaining single-copy polymerase chain reaction products in the expected size range proved to be difficult, and only 24% of the primer pairs tested produced polymorphic single-locus markers. We obtained five highly variable loci with 16 to 41 alleles and an observed heterozygosity ranging from 0.29 to 0.81. Amplifications have also been obtained from the earliest postlarval stage. These highly informative markers will allow studies of the population structure of P. koreni at fine spatial and temporal scales. PMID- 10804245 TI - Comparative Analysis of Two Populations of the Brittle Star Amphiura filiformis (Echinodermata: Ophiuroidea) with Different Life History Strategies Using RAPD Markers. AB - Random amplified polymorphic DNA (RAPD) analysis was applied to individuals of Amphiura filiformis sampled from two geographic locations that have previously been reported to have different life history strategies. Of thirty-one 10-mer RAPD primers screened initially, four were chosen and used in a comparative analysis of A. filiformis individuals collected from Galway Bay (Ireland) and Concarneau Bay (France). The results show much variation within A. filiformis populations. Although there are some rare alleles particular to each population, overall the populations were not genetically differentiated with the methods employed. Analysis of molecular variance (AMOVA) showed that a minimum of 93% of genotypic variance occurred among individuals within populations. The statistical significance of this pattern was supported by permutation tests. F(ST) values were not significantly different from and UPGMA cluster analyses based on three distance metrics did not separate the two populations. PMID- 10804257 TI - Subcritical endemic steady states in mathematical models for animal infections with incomplete immunity. AB - Many classical mathematical models for animal infections assume that all infected animals transmit the infection at the same rate, all are equally susceptible, and the course of the infection is the same in all animals. However for some infections there is evidence that seropositives may still transmit the infection, albeit at a lower rate. Animals can also experience more than one episode of the infection although those who have already experienced it have a partial immune resistance. Animals who experience a second or subsequent period of infection may not necessarily exhibit clinical symptoms. The main example discussed is bovine respiratory syncytial virus (BRSV) amongst cattle. We consider simple models with vaccination and homogeneous and proportional mixing between seropositives and seronegatives. We derive an expression for the basic reproduction number, R(o), and perform an equilibrium and stability analysis. We find that it may be possible for there to be two endemic equilibria (one stable and one unstable) for R(o)<1 and in this case at R(o)=1 there is a backwards bifurcation of an unstable endemic equilibrium from the infection-free equilibrium. Then the implications for control strategies are considered. Finally applications to Aujesky's disease (pseudorabies virus) in pigs are discussed. PMID- 10804258 TI - A delay-differential equation model of HIV infection of CD4(+) T-cells. AB - A.S. Perelson, D.E. Kirschner and R. De Boer (Math. Biosci. 114 (1993) 81) proposed an ODE model of cell-free viral spread of human immunodeficiency virus (HIV) in a well-mixed compartment such as the bloodstream. Their model consists of four components: uninfected healthy CD4(+) T-cells, latently infected CD4(+) T cells, actively infected CD4(+) T-cells, and free virus. This model has been important in the field of mathematical modeling of HIV infection and many other models have been proposed which take the model of Perelson, Kirschner and De Boer as their inspiration, so to speak (see a recent survey paper by A.S. Perelson and P.W. Nelson (SIAM Rev. 41 (1999) 3-44)). We first simplify their model into one consisting of only three components: the healthy CD4(+) T-cells, infected CD4(+) T-cells, and free virus and discuss the existence and stability of the infected steady state. Then, we introduce a discrete time delay to the model to describe the time between infection of a CD4(+) T-cell and the emission of viral particles on a cellular level (see A.V.M. Herz, S. Bonhoeffer, R.M. Anderson, R.M. May, M.A. Nowak [Proc. Nat. Acad. Sci. USA 93 (1996) 7247]). We study the effect of the time delay on the stability of the endemically infected equilibrium, criteria are given to ensure that the infected equilibrium is asymptotically stable for all delay. Numerical simulations are presented to illustrate the results. PMID- 10804259 TI - Keratinocyte growth factor signalling: a mathematical model of dermal-epidermal interaction in epidermal wound healing. AB - A wealth of growth factors are known to regulate the various cell functions involved in the repair process. An understanding of their therapeutic value is essential to achieve improved wound healing. Keratinocyte growth factor (KGF) seems to have a unique role as a mediator of mesenchymal-epithelial interactions: it originates from mesenchymal cells, yet acts exclusively on epithelial cells. In this paper, we study KGF's role in epidermal wound healing, since its production is substantially up-regulated after injury. We begin by modelling the dermal-epidermal signalling mechanism of KGF to investigate how this extra production affects the signal range. We then incorporate the effect of KGF on cell proliferation, and using travelling wave analysis we obtain an approximation for the rate of healing. Our modelling shows that the large up-regulation of KGF post-wounding extends the KGF signal range but is above optimal for the rate of wound closure. We predict that other functions of KGF may be more important than its role as a mitogen for the healing process. PMID- 10804260 TI - Effect of aggregating behavior on population recovery on a set of habitat islands. AB - We consider a single-species model which is composed of several patches connected by linear migration rates and having logistic growth with a threshold. We show the existence of an aggregating mechanism that allows the survival of a species which is in danger of extinction due to its low population density. Numerical experiments illustrate these results. PMID- 10804261 TI - Weakly coupled hyperbolic systems modeling the circulation of FeLV in structured feline populations. AB - Global existence and regularity results are provided for weakly coupled first order hyperbolic systems modeling the propagation of the Feline Leukemia Virus (FeLV), a retrovirus of domestic cats (Felis catus). In a simple example we find a threshold parameter yielding endemic stationary states. PMID- 10804262 TI - Ozone and the lung: a sensitive issue. AB - Ozone is a powerful oxidant and toxic air pollutant. As a gaseous pollutant, its primary target tissue is the lung and breathing slightly elevated concentrations of ozone results in a range of respiratory symptoms. These include decreased lung function and increased airway hyper-reactivity in 10-20% of the healthy population. Moreover, those with conditions such as asthma and chronic obstructive pulmonary disease (COPD) generally experience an exacerbation of their symptoms. Together, these observations suggest that certain individuals are particularly susceptible to this oxidant gas. The primary goal of this review is to examine the basis of this increased sensitivity. Ozone is a highly reactive gas that is consumed by reactive processes on reaching the first interface in the lung, the lung lining fluid compartment. Reactions between ozone and antioxidants tend to dominate in this compartment and these are generally thought of as beneficial, or protective interactions. In those instances when ozone reacts with other substrates in lung lining fluid such as protein or lipid, secondary oxidation products arise which transmit the toxic signals to the underlying pulmonary epithelium. The rules that govern the balance between beneficial and detrimental interactions in the lung lining fluid compartment are not well established but these may contribute, in part, to sensitivity. On reaching the lung surface, secondary oxidation products arising from ozone initiate a number of cellular responses. These include cytokine generation, adhesion molecule expression and tight junction modification. Together, these responses lead to the influx of inflammatory cells to the lung in the absence of a pathogenic challenge. Moreover, lung permeability is increased and oedema develops. The nature and extent of these responses are variable and often not related within an individual. Thus, although an improved appreciation of the general mechanism of action of ozone has been attained in recent years, the basis for individual susceptibility is still unclear. PMID- 10804263 TI - Least-effort pathways?: a GIS analysis of livestock trails in rugged terrain. AB - Livestock trails frequently evolve in pastures when plant growth or establishment cannot keep pace with vegetation disturbance. In some instances, man-made trails are established in rangeland settings to encourage uniform use of forages or facilitate livestock passage through dense vegetation or across rugged terrain. A long-term assumption has been that livestock establish pathways of least resistance between frequented areas of their pastures, but this hypothesis has never been tested. We mapped cattle trails in three 800+ ha pastures with global positioning units. A geographic information system (GIS) helped quantify characteristics of trails and the landscape and was used to plot least-effort pathways between water sources and distant points on selected trails in the pastures. Characteristics of the cattle trails and least-effort pathways were compared to test the hypothesis that cattle develop least-effort routes of travel in rugged terrain. The mean slope of the three pastures was 13.5%, and the average slope of the topography traversed by the cattle trails was 8%. The slope of the trails was reduced to 5.2% by selection of cross-slope routes. When we compared the characteristics of 10 selected cattle trails and least-effort pathways generated by our GIS, the cattle trails were 11% shorter (P=0.046) than the least-effort pathways, and the topography traversed by cattle had a gradient about 1% less than the least-effort pathways (P=0.02). The slope of the selected trails (5.5%) and pathways (5.6%) were similar (P=0.74), however. Analyses of values extracted from cost surfaces indicated that, on the average, 183 units of effort were needed to traverse the trails and 170 units of effort expended to traverse the least-effort pathways (P=0.07). These data support the hypothesis that cattle establish least-effort routes between distant points in rugged terrain and suggest that GIS software may be useful in designing systems of livestock trails in extensive settings. PMID- 10804264 TI - Mixing at young ages reduces fighting in unacquainted domestic pigs. AB - Under normal farming practices, piglets from different litters are often mixed around the time of weaning, and a high incidence of fighting and minor injuries often occur. The aim of this experiment was to determine the effect of age on the incidence of fighting in piglets mixed before weaning, at different ages between 5 and 26 days. We found no significant relationship between age and the likelihood that a pair of piglets would fight during the first 75 min after mixing. However, the duration of the first fight observed increased from 101+/-38 s at 5 days to 621+/-278 s at 26 days, mainly because of higher levels of unretaliated harassment and resting during the bouts. Younger pigs also showed 80% fewer injuries from the fighting. The results suggest some potential welfare advantage to allowing litters to mix at younger ages. PMID- 10804265 TI - Sexual performance of twin ram lambs and the effect of number and sex of contemporary siblings. AB - The objectives of this investigation were to determine the degree of correspondence in the sexual performance of twin male sheep and to compare the sexual performance of ram lambs born co-twin to another male (M-M), co-twin to a female (M-F) and as single (S) offspring. Individual ram lambs (N=117), including 13 M-M twin sets, were exposed to four estrous ewes for 30 min at 7-day intervals over a 4-week period at 8 to 9 months of age.Frequencies of ejaculation (serving capacity) and mounts without ejaculation were recorded. M-M males averaged 2.9 ejaculations and 13.1 mounts per test. Individuals within 9 of the 13 M-M twin sets (69%) differed by an average of 0.5 or fewer ejaculations per test. Although variability within M-M twin sets was very low, variability between the 13 M-M twin sets was also low (coefficient of variation=10.6%). Consequently, frequencies of both ejaculation and mounting (without ejaculation) within the 13 twin sets were not correlated (P=0.50 and P=0.10, respectively). When the progeny of individual sires were compared, mounting frequencies were correlated (P<0.02) but not ejaculation rates (P=0.72). M-M, M-F and S ram lambs did not differ for either ejaculation frequency (P=0.26) or mounts without ejaculation (P=0.98). It was concluded that the sexual performance of related individuals is likely to be very similar and that the number and sex of contemporary siblings does not necessarily influence rates of mounting and ejaculation. PMID- 10804266 TI - T-maze behaviour in broiler chicks is not sensitive to right-left preferences, test order or time-of-day. AB - There is substantial individual variation in the time taken by broiler chicks to traverse a T-maze and thereby reinstate visual contact with their companions. Chicks completing this task quickly (high performance, HP) subsequently grew faster in the laboratory and on farm, exhibited greater sociality, and showed less pronounced adrenocortical responses to a partial water immersion stressor than did their slower (low performance, LP) counterparts [Marin, R.H., Jones, R.B., 1999. Latency to traverse a T-maze at 2 days of age and later adrenocortical responses to an acute stressor in domestic chicks. Physiol. Behav. 66, 809-813.; Marin, R.H., Arce, A., Martijena, I.D., 1997. T-maze performance and body weight relationship in broiler chicks. Appl. Anim. Behav. Sci. 54, 197 205.; Jones, R.B., Marin, R.H., Garcia, D.A., Arce, A., 1999. T-maze behaviour in domestic chicks: a search for underlying variables. Anim. Behav. 58, 211-217.]. Given its simplicity, rapidity and non-invasiveness, the T-maze test might represent a commercially attractive selection criterion for future breeding programmes if this behavioural trait exhibits sufficient genetic variability. However, it is first necessary to ensure that performance in the T-maze is not sensitive to potentially confounding variables, such as existing preferences to turn right or left at the junction of the maze, the order of testing, or the time of day. In the present study, 240 newly hatched, mixed-sex broiler chicks (Cobb) were randomly allocated to 12 groups of 20 upon receipt. When they were 2 days of age a group of 20 chicks was placed in the brood area of each of two T-mazes at 0830 h; the brood areas were positioned on either the right or the left sides of the mazes. After acclimatisation, one chick from each group was placed in the isolation chamber of the T-maze and we recorded the time it took to traverse the maze. This procedure was repeated until all 20 chicks and all 12 groups had been tested. The locations of the brood areas were rotated after each block of 20 tests. Testing was completed in the same day and two groups were tested at each of the following times: 0900, 1030, 1200, 1330. 1500, and 1630 h. Analyses of variance (ANOVA) revealed no detectable effects of: positioning the brood area on the right or the left side of the T-maze (F(1,200)=0.06, P<0.80), test order (F(19,200)=0.48; P<0.96), or time-of-day (F(5,234)=0.44; P<0.81). These findings strongly suggest that the future classification and possible selection of broiler chicks according to their T-maze responses are unlikely to be confounded by right left preferences, test order, or the time of day. PMID- 10804267 TI - Effect of manipulating feathers of laying hens on the incidence of feather pecking and cannibalism. AB - Feather pecking is a problem in commercial laying hens, particularly in loose housing systems, where many hens can be affected by only a few feather peckers. In addition, feather pecking can become an even larger problem if it spreads throughout the flock. There are several possible ways that feather pecking may spread. The simplest way is that one hen may damage the feathers of a hen, and another hen may find the damaged feathers an attractive pecking target. The aim of this experiment was to determine if damaged feathers were feather-pecked more than undamaged feathers on the same body area, and to determine whether some types of feather-body area manipulations were preferred over others as pecking stimuli. Manipulations involved damaging the feathers on the rump, tail or belly of different hens, with two or three levels of severity of manipulation at each body area. Sixteen groups of 11 Lohmann Brown hens between 26 and 28 weeks were observed with the recipient, the feather pecker and the body area that was pecked all being recorded. The feather pecks were classified separately as either gentle or severe. Damaged feathers received significantly more severe feather pecks than undamaged feathers. There were also more gentle feather pecks to damaged feathers, although this did not reach statistical significance. The feather-body area manipulations that received the greatest number of severe feather pecks were the tail feathers when they were cut very short, the rump feathers when they were trimmed, and the rump when feathers were removed. These results support the suggestion that feather pecking does indeed spread through flocks by damaged feathers becoming an attractive target for feather-pecking behaviour. An unexpected result of performing the feather manipulations was an outbreak of cannibalism in half of the experimental groups. Even though there was no visible damage to the skin of the hens after having the feathers manipulated, 13 of the 16 attacked hens were wounded on the part of the body where the feathers had been damaged in some way. PMID- 10804268 TI - Early locomotor behaviour in genetic stocks of chickens with different growth rates. AB - Reduction in exercise increases the occurrence of lameness in meat-type chickens. Locomotor activity is dramatically reduced during the finishing period in chickens from fast-growing genetic types compared to slow-growing genetic types, but it is not known whether this difference is already present during the starting period and may be influenced by genetic factors. In order to define the effect of genetic origin on early locomotor behaviour, exercise was compared from 1 to 22 days of age in two meat-type chicken stocks differing in growth rate: male broilers (B) which grow fast and are often lame, and male "label rouge" chickens (L) which grow slowly and are rarely lame.Time budget (lying, standing, drinking, eating, walking) was measured by scanning in six repetitions of five birds (density=2.5 birds/m(2)) at 1, 8, 15 and 17 days of age. Standing bouts were analysed by focal sampling at 2-3, 6-7, 13-14 and 20-21 days of age.B chicks spent less time standing than L chicks at 15 days of age (B=13+/-2%, L=24+/-1%, P<0.01) and 17 days of age, and spent more time lying at 17 days of age (B=73+/ 3%, L=60+/-4%, P<0.05).The major part (74%) of the total active time observed by focal sampling was linked to feeding activity. At 2 and 3 days, the activity of B chicks was half that of L chicks during standing bouts (duration of walking per bout: 19+/-4 s for B; 45+/-4 s for L, P<0.05). The activity observed by focal sampling during non-feeding bouts at 20-21 days was significantly correlated with the corresponding data recorded at 2-3 days in the same chicks in the B stock but not in the L stock.We concluded that (1) both B and L genetic stocks have the same overall activity during the first 3 days of age (scanning) but they exhibit different organisation and composition of standing bouts (focal sampling). (2) Genetic factors are probably involved in the expression of locomotor behaviour in very young chicks. (3) The correlations between the levels of activity at early and later ages suggest that selection of young mobile broiler chicks might increase activity at a later age and might therefore reduce the occurrence of leg abnormalities. PMID- 10804269 TI - Night-time roosting in laying hens and the effect of thwarting access to perches. AB - Free-living hens roost on branches in trees at night, and laying hens in aviary systems or cages provided with perches also make extensive use of these for night time roosting. It is therefore suggested that roosting on perches is important to the hens and that domestic hens should be provided with perches in order to promote welfare. However, no study has addressed the question of motivation for roosting. In the present experiment, we studied undisturbed roosting behaviour and the reaction of commercial laying hens when roosting on perches was thwarted. Fifty-two adult hens (Lohmann Selected Leghorn, LSL) were kept in two groups of 26 hens in litter pens with perches at heights of 23, 43 and 63 cm. Behaviour was observed for 60 min starting at lights-off, registering the number of hens on each perch level. The hens started to get onto the perch immediately and within 10 min after lights-off, more than 90% of the hens were on the perch. All hens roosted close together on the top perch. In a second experiment, 24 hens were kept in eight groups of three birds each in experimental pens equipped with perches. Birds were tested in four different situations: (1) the pen unchanged (Base), (2) the perch covered with plexiglass (PCov), (3) the perch removed (PRem) and (4) the unchanged pen (Post). The order of PCov and PRem alternated between groups in a balanced manner and all groups of birds experienced all four treatments. The hens were observed for 60 min from lights-off using focal sampling. For comparisons, the Post treatment served as the control. In the treatments where perching was not possible, the hens spent less time sitting (p=0.042), and also tended to spend more time standing (p=0.06), than in the control. Furthermore, the hens moved more (p=0.042) when the perch was inaccessible, and when the perch was visible but inaccessible they also showed more attempts to take off (p=0.042). These findings can be interpreted as increased frustration and/or exploration, probably to find an alternative roosting site. Together with the high use of perches for night-time roosting under undisturbed conditions, these results indicate that laying hens are motivated to perch and imply that hens kept under conditions where perching is not possible may experience reduced welfare. PMID- 10804270 TI - Differences in food preferences between individuals and populations of domestic cats Felis silvestris catus. AB - The food preferences of 64 cats, of which 28 were domestic pets and 36 were free ranging animals on three farms, were tested using five food types; a sixth type was also tested on the farm cats. Information was also gathered on the background diet of the house cats (individually) and of each farm cat colony. Consistent differences were found between the predefined groups of cats in their preferences for three foods, Hard Dry (HD), Canned Meat (CM) and Raw Beef (RB), the greatest differences being between house cats and farm cats as a whole. The three colonies of farm cats showed distinct differences in preference, for RB and/or CM, and for the sixth food, Soft Dry (SD). Differences between farms could be explained by a reduced preference for items similar to major components of the background diet of each colony, i.e. selection in favour of foods that were temporally rare. Differences between house and farm cats, and within the house cats, could not be explained in this way; the house cats were neophobic towards RB, and the farm cats ate little of the HD food, possibly because they found it difficult to ingest. PMID- 10804271 TI - The effect of dose and route of oestradiol benzoate administration on plasma concentrations of oestradiol and FSH in long-term ovariectomised heifers. AB - Oestradiol (E(2)) suppresses FSH and affects follicle wave dynamics in cattle. However, neither the optimum dose of ODB required to suppress FSH nor the effect of route of ODB administration on blood concentrations of E(2) are known; hence, the aim of this experiment was to answer these questions. Ovariectomised heifers received Progesterone Releasing Intravaginal Device (PRID) for 7 days, and 4 days later heifers received one of eight ODB treatments at second PRID insertion as follows; (1) 0.0 mg (Control; n=3), (2) 0.5 mg (n=4), (3) 1.0 mg (n=4), (4) 2.5 mg (n=6), (5) 5.0 mg (n=4), (6) 10. 0 mg (n=4), (7) 5.0 mg (n=4), and (8) 10.0 mg (n=5). For treatments 2-6 inclusive, ODB was administered intramuscularly in oil, while for treatments 7 and 8, the ODB in powder form was administered topically in the vagina by gelatine capsule attached to the PRID. Blood samples were collected every 6 h for the first 48 h, every 12 h for the next 48 h, and twice daily for a further 6 days. The interval from ODB administration to peak E(2) concentration was similar (P0.05) for treatments 2-6 where ODB was administered intramuscularly (mean 13.4+/-1.24 h), and was longer (P<0.05) for the intravaginal capsule treatments (mean 25.5+/-2.84 h). Plasma concentrations of E(2) increased with increasing intramuscular dose of ODB injected, (plasma E(2)= 0.237+16.109 (dose)-0.74 (dose)(2), R(2)=0.75; P<0.05). Peak plasma concentrations of E(2) following the 5- and 10-mg capsules were similar to each other and to those following the 0.5-mg injection (P0.05), but were lower than concentrations obtained following injection of 1.0-5.0 mg (P<0.05). Across all treatments, both the maximum percentage decline in FSH and the interval to FSH nadir were related to the peak plasma concentrations of E(2) (maximum % decline in FSH=11.17+1.564 (peak E(2))-0.009 (peak E(2))(2), R(2)=0.75; P<0.01), (hours to FSH nadir=10.628+1.486(hours to peak E(2))-0.0282(hours to peak E(2))(2), R(2)=0.22; P<0.05). Concentrations of FSH increased as E(2) declined from its peak value, irrespective of maximum value achieved. It was concluded that the intramuscular administration of ODB in oil to ovariectomised heifers given a PRID results in higher plasma concentrations of E(2) and causes a greater reduction in FSH than administration topically by intravaginal gelatine capsule. E(2) transiently suppresses FSH in ovariectomised heifers, and the magnitude of the suppression is dose-dependent; however FSH concentrations begin to increase 1-2 days after ODB administration while concentrations of E(2) were declining but still high. PMID- 10804272 TI - Improved development of DNA-injected bovine embryos co-cultured with mouse embryonic fibroblast cells. AB - The in vitro development of DNA-injected bovine zygotes, produced in vitro, was compared when cultured with or without mouse embryonic fibroblasts (MEF). The in vivo viability of the embryos produced in these in vitro culture systems was assessed by single or double transfer to recipients taken to term. For these experiments, in vitro fertilized oocytes were not injected (Experiment 1) or were injected with pBL1 gene (Experiment 2) and then cultured for 2 days in CR1aa medium supplemented with 3 mg/ml BSA at 38.5 degrees C in a humidified atmosphere of 5% CO(2) in air. Embryos that developed to the 4- to 8-cell stage at the end of this period were randomly assigned to the two cultured systems and cultured for a further 5 days in groups of 10 to 15 embryos in 0.75 ml medium. These two culture systems were CR1aa medium alone or co-culture with MEF in CR1aa medium supplemented with 10% fetal bovine serum (FBS). Every 48 h, 0.5 ml of the medium was replaced with fresh CR1aa medium and at Day 5 of culture, both media were supplemented by the addition of 5.56 mM glucose and 1x GMS-X supplement solutions. Results were assessed as morphological development of the embryos and data were analyzed by Chi-square test or Student's t-test. The development rate of in vitro fertilization (IVF)-derived embryos co-cultured with MEF (24.4%, 49/201) was significantly higher than those cultured alone (14.4%, 28/194; P<0.05) in Experiment 1. There was a similar difference between the treatments in the proportions of embryos which reached the hatching stage or hatched (10.9%, 22/201 vs. 4.1%, 8/194, respectively; P<0.05). DNA-injected embryos co-cultured with MEF (13.7%, 28/205) showed a higher developmental rate than that of the embryos cultured without MEF (6.7%, 13/193; P<0.05) in Experiment 2. Following the transfer to recipients of one or two DNA-injected blastocysts, the pregnancy rates for two culture systems were similar (MEF co-culture 27.4%, 23/84; CR1aa culture 24. 2%, 16/66). However, the numbers of calves born alive from these pregnancies were higher on the MEF co-culture group (82.6%, 19/23) than the CR1aa culture group (56.2%, 9/16). It was concluded that in vitro embryo development to the blastocyst stage and subsequent in vivo development to term of DNA-injected bovine embryos was improved in comparison to culture in CR1aa alone when the last 5 days of in vitro culture were in a MEF co-culture system. PMID- 10804273 TI - Effects of ammonia during different stages of culture on development of in vitro produced bovine embryos. AB - The effects of various concentrations of ammonia in the media during in vitro fertilization (IVF), culture (IVC), and throughout maturation (IVM), IVF, and IVC were evaluated using a randomized complete block design. Ammonia was added to the media at various concentrations during IVF (experiment 1), during IVC (experiment 2), and throughout IVM, IVF, and IVC (experiment 3). In the first experiment, there was a significant (P<0.05) increase in embryos developed to blastocyst, and to expanding and hatching blastocyst, in IVF media containing moderate concentrations of ammonia compared with that in the IVF control media. In the second experiment, ammonia in the IVC media increased (P<0.05) the proportion of degenerate ova and decreased (P<0.05) the proportion of ova that developed to blastocysts. In experiment 3, cleavage rates tended (P=0.06) to be greater for control groups than for treatment groups. The proportion of ova developing to morula was greater (P<0.05) in media containing moderate concentrations of ammonia than that in the control groups. These results indicate that the effect of ammonia on development of preimplantation bovine embryos depends on the concentration of ammonia and the stage of development when exposure to ammonia occurs. PMID- 10804274 TI - Effect of buffering systems on post-thaw motion characteristics, plasma membrane integrity, and acrosome morphology of buffalo spermatozoa. AB - This study was carried out to identify the suitable buffer for cryopreservation of buffalo semen. Semen was collected with artificial vagina (42 degrees C) from four buffalo bulls. Split pooled ejaculates (n=5), possessing more than 60% visual sperm motility, were extended at 37 degrees C either in tri-sodium citrate (CITRATE), Tris-citric acid (TCA), Tris-Tes (TEST) or Tris-Hepes (HEPEST). Semen was cooled to 4 degrees C in 2 h, equilibrated at 4 degrees C for 4 h, filled in 0.5 ml straws and frozen in a programmable cell freezer before plunging into liquid nitrogen. Thawing of frozen semen was performed after 24 h at 37 degrees C for 15 s. Sperm motion characteristics, plasma membrane integrity, and acrosome morphology of each semen sample were assessed by using computer-assisted semen analyzer (CASA), hypo-osmotic swelling (HOS) assay, and phase-contrast microscope, respectively. Analysis of variance revealed that percent post-thaw visual motility tended (P=0.07) to be higher in HEPEST (61.0+/-2.9) and lowest in CITRATE (48.0+/-2.5). Computerized motility did not vary due to buffering system. Percent post-thaw linear motility tended (P=0.09) to be higher in TCA (78.2+/ 5.5) and lower in TEST (52.0+/-6.9). Circular motility (%) was significantly lower (P<0.05) in TCA (11.6+/-2.8) and higher in TEST (29.8+/-5.6). Curvilinear velocity (microm s(-1)) was lower (P<0.05) in TCA (69.4+/-2.0) than in CITRATE (79.0+/-5.8), TEST (87. 2+/-1.6) and HEPEST (82.6+/-3.0). Lateral head displacement (microm) was lowest (P<0.05) in TCA (1.7+/-0.2) and highest in TEST (3.7+/-0. 6). Plasma membrane integrity and normal acrosomes of buffalo spermatozoa did not differ due to buffering system and averaged 40. 0+/-2.7% and 61.4+/-4.6%, respectively. Based upon lower circular motility, curvilinear velocity, and lateral head displacement, it is concluded that post-thaw quality of buffalo semen can be improved using the Tris-TCA buffering system. PMID- 10804275 TI - Genetic influences on reproduction of female red deer (Cervus elaphus) (1) seasonal luteal cyclicity. AB - This study compared the onset and duration of the breeding season of female red deer (Cervus elaphus scoticus) and its hybrids with either wapiti (Cervus elaphus nelsoni) or Pere David's (PD) deer (Elaphurus davidianus). In Trial 1 (1995), adult red deer (n=9), F1 hybrid wapiti x red deer (n=6) and maternal backcross hybrid PD deer x red deer (i.e., 14 PD; n=9) were maintained together in the presence of a vasectomised red deer stag for 12 months. They were blood-sampled daily or three times weekly so that concentration profiles of plasma progesterone could be used to identify the initiation, duration and cessation of luteal events. There was clear evidence of luteal cyclicity between April and September, with the transition into breeding associated with an apparent silent ovulation and short-lived corpus luteum (i.e., 6-12 days) in every hind. A significant genotype effect occurred in the mean time to first oestrus (P<0.05), with wapiti hybrids and 14 PD hybrids being 9 and 5 days earlier than red deer. Between six and nine oestrous cycles were exhibited by each hind, with no difference in mean cycle length (19.5-19.6 days) between genotypes (P0.10). The overall length of the breeding season was significantly longer for wapiti hybrids (143 days) than for either red deer (130 days) and 14 PD hybrids (132 days, P<0.05). In Trial 2 (1998), adult red deer (n=5), 14 PD hybrids (n=5) and F(1) PD x red deer hybrid (n=5) hinds were maintained together from mid-February (late anoestrus) to early May, in the presence of a fertile red deer stag from 1 April. Thrice-weekly blood sampling yielded plasma progesterone profiles indicative of the onset of the breeding season. Again, there was a significant genotype effect on the mean time to first oestrus (P<0. 05), with F(1) PD hybrids and 14 PD hybrids being 13 and 5 days earlier than red deer. However, conception dates were influenced by the timing of stag joining, and were not significantly different between genotypes. The results indicate genetic effects on reproductive seasonality. However, seasonality observed for PD x red deer hybrids more closely approximated that of red deer than PD deer. PMID- 10804276 TI - Genetic influences on reproduction of female red deer (Cervus elaphus) (2) seasonal and genetic effects on the superovulatory response to exogenous FSH. AB - This study evaluated the influences of seasons and genotype on the superovulatory response to a standardised oFSH regimen in red deer (Cervus elaphus scoticus) and its hybrids with either wapiti (C.e. nelsoni) or Pere David's (PD) deer (Elaphurus davidianus). Adult red deer (n=9), F(1) hybrid wapiti x red deer (n=6), and maternal backcross hybrid PD x red deer (i.e., 14 PD hybrid; n=9) were kept together in the presence of a vasectomised stag for 13 months. At 6 weekly intervals, all hinds received a standardised treatment regimen used routinely to induce a superovulatory response in red deer hinds, with 10 consecutive treatments spanning an entire year. This involved synchronisation with intravaginal progesterone devices and delivery of multiple injections of oFSH (equivalent to 72 units NIH-FSH-S(1)). Laparoscopy to assess ovarian response was performed 6-7 days after the removal of the devices. Both season and genotype had significant effects on ovulation rate (OR) and total follicular stimulation (TFS) (P<0.05). For all the three genotypes, ovarian responses were highest from March to November (breeding season) and lowest in the period from December to January, inclusive. Mean OR for red deer hinds ranged from 3.7 to 1.8 during the breeding season, with no observable trend. All red deer hinds were anovulatory during December and January. A similar pattern occurred for 14 PD hybrids, although mean OR during the breeding seasons were twofold lower than for the red deer. For F(1) wapiti hybrids, the first two treatments in March and April resulted in the highest mean OR observed (15.6 and 11.7, respectively). Thereafter, mean values ranged between 6.3 and 4.7 for the remainder of the breeding season. Furthermore, mean OR of 3.0 and 0.5 were recorded in December and January, respectively. For the red deer and F(1) wapiti hybrids, between-hind variation in OR was not randomly distributed across the treatment dates, indicating that the individuals varied significantly in their ability to respond to oFSH, at least within a given season.In conclusion, the study has shown that relative to red deer, F(1) wapiti hybrid hinds exhibit a higher sensitivity to oFSH, whereas 14 PD hybrid hinds have a lower sensitivity. However, individual variation within genotype was very marked. A seasonal effect was apparent for all genotypes, although some F(1) wapiti hybrid hinds exhibited ovulatory responses throughout the year. PMID- 10804277 TI - Comparing ovulation synchronization protocols for artificial insemination in the scimitar-horned oryx (Oryx dammah). AB - Ovarian response and pregnancy success in scimitar-horned oryx (n=28) were compared, following treatment with two synchronization protocols and fixed-time artificial insemination (AI) with frozen-thawed semen. Each oryx received two injections of 500 microg of prostaglandin-F(2alpha) analogue (PGF(2alpha)-only) 11 days apart, and half received PGF(2alpha) in combination with an intravaginal progesterone-releasing device (CIDR11+PGF(2alpha)). Semen was collected by electroejaculation from anaesthetised adult oryx and cryopreserved. Anaesthetised females were transcervically inseminated 56.0+/-1.1 h (+/-S.E.M.) after PGF(2alpha) injection and/or device withdrawal using 28.0+/-1.5x10(6) motile thawed sperm. Ovarian endocrine response was monitored in 20 females by analysing faecal oestrogen and progesterone metabolites. Periovulatory oestrogen peaks were detected in 19/20 (95%) females after synchronization. There were no between treatment differences in oestrogen concentrations or peak characteristics (P0.05). Luteal development after synchronization was delayed in half the progesterone treated (CIDR11+PGF(2alpha)) females, and faecal progestin excretion profiles indicated that the ovulatory follicle associated with synchronization either failed to ovulate or to fully lutenise. Pregnancy was diagnosed by ultrasonography and/or rectal palpation and was monitored by faecal progestin excretion. More (P=0. 013) pregnancies resulted from the PGF(2alpha)-only treatment (37.5%, 5/14) than from the CIDR11+PGF(2alpha) treatment (0/14), and four healthy scimitar-horned oryx calves were born, three after gestation intervals of 247 days and one after 249 days. PMID- 10804278 TI - Levels of progesterone and changes in prostaglandin F(2alpha) release during luteolysis and early pregnancy in llamas and the effect of treatment with flunixin meglumine. AB - The secretory patterns of progesterone in relation to concentrations of 15 ketodihydro-PGF(2alpha) (PGFM) during the period of luteolysis or of maternal recognition of pregnancy were determined in the blood of llamas mated either with an intact or a vasectomized male. The ability of flunixin meglumine (FM) to postpone luteolysis in non-pregnant llamas was investigated by injecting the drug intravenously every 6 h at a dose of 2.2 mg/kg from days 6 to 12 post-copulation into a group of non-pregnant llamas. A pulsatile pattern of prostaglandin release was recorded during luteolysis in non-pregnant llamas, giving further support to the hypothesis that PGF(2alpha) is the luteolytic agent in llamas. The mean number of peaks per animal rose from 0.3 on day 7 to 3.8 on day 10 and then declined to 1.1 on day 12 with corresponding mean peak amplitude changing from 465 to 1234 and 566 pmol l(-1), respectively. In pregnant llamas, prostaglandin pulsatile release also occurred. The mean number of peaks per animal rose from 0.4 on day 7 to 0.8 on day 10 and then declined to 0.2 on day 11 and 0.6 on day 12, with corresponding mean peak amplitude changing from 494 to 676, 388 and 547 pmol l(-1), respectively. The transient decrease and subsequent recovery in progesterone concentrations was observed to occur in connection with prostaglandin release during early pregnancy. Oestradiol-17beta plasma peak concentrations attained after luteolysis were significantly higher than those recorded in early pregnant animals (around 30 pmol l(-1) and ll pmol l(-1)). Concentrations of PGFM decreased rapidly after the first administration of FM and remained low throughout the first 2 days of treatment. Thereafter, pulsatile release of prostaglandins started, and luteolysis proceeded; but a delay of 1-1.5 days in the progesterone decline was observed. Thus, it might be suggested that a higher dose and/or a more intensive injection schedule is required in llamas than in other ruminants to prevent luteolysis. PMID- 10804279 TI - Influence of sequence duration and number of electrical pulses upon rabbit oocyte activation and parthenogenetic in vitro development. AB - Electroactivation of in vivo mature young rabbit oocytes was investigated here. The effects of four or eight electrical pulse treatment over 90, 150 or 270 min upon oocyte activation frequency and type, and even upon their subsequent in vitro development, were studied. The lowest activation frequency was observed after applying four-pulses over 90 min (54%). However, extending four-pulse treatment duration over 150 or 270 min induced more oocytes to activate (from 84% to 100%), as did the eight pulsing treatments (from 91% to 97%). With eight pulses, extending treatment duration improved the normal activation rates (from 47% to 76%; P<0.05). Nevertheless, the haploid morulae and blastocyst rates decreased significantly with extended eight pulsing treatment duration (morulae: from 94% to 41% and blastocysts: from 31% to 0%). PMID- 10804281 TI - Corrigendum to "Manual collection and characterization of semen from asian elephants" PMID- 10804280 TI - Natural and radiation-induced degeneration of primordial and primary follicles in mouse ovary. AB - The present study deals with the morphological changes of the degenerating primordial and primary follicles induced by gamma-radiation. Prepubertal female mice of 3 weeks old ICR strain were gamma-irradiated with the dose of LD(80(30)) (8.3 Gy). The ovaries were collected at 3, 6 and 12 h after irradiation. The largest cross-sections were prepared by histological semithin sections for microscopical observations. The ratio (%) of normal to atretic follicles decreased with time after the irradiation in primordial follicles and in primary follicles as well. At 6 h after irradiation, the number of degenerated primordial follicles increased. Germinal vesicles disappeared and lipid droplets increased in number. Granulosa cells became round in shape and apoptotic cells started to appear. The ooplasmic membrane was not recognizable. The ratio of normal to atretic primordial follicles in the control group was 62.5. Then it became lower with time after the irradiation. It went down to 51.6, 49.0, 11.1 and 7.1 at 0, 3, 6 and 12 h, respectively. The ratio of normal to atretic primary follicles in the control mouse ovary was 81.3. It was 80.0, 75.0, 45.5 and 33. 3 at 0, 3, 6 and 12 h after irradiation, respectively. It is concluded that the ionizing radiation acutely induces the degeneration of primordial and primary follicles. The pattern of degeneration is one of the following: (1) apoptosis of one or more granulosa cells with a relatively intact oocyte, (2) apoptosis of an oocyte with intact follicle cells, or (3) apoptotic degenerations of both kinds of cells. These results can provide morphological clues for the identification of the degenerating primordial and primary follicles in normal and irradiated mouse ovaries. PMID- 10804282 TI - Cationic liposome conjugation to recombinant adenoviral vector reduces viral antigenicity. AB - Adenoviral (Ad) vectors are commonly used in gene therapy trials because of their efficiency in gene transfer. However, their use is limited by immune responses that reduce transgene expression and decrease the efficacy of repeated vector administration. In this study, we demonstrated that conjugation of Ad vector with our novel cationic liposomes could reduce viral antigenicity in vivo. Mice subcutaneously injected with liposome-conjugated Ad vector showed a 6.5-fold reduction of anti-Ad antibodies with neutralizing activity, compared to those with unconjugated Ad vector. Interestingly, we also found that the conjugated vector is less susceptible to inactivation by neutralizing antibodies in vitro and in vivo. Our results suggest that liposome conjugation reduces viral antigenicity, shields vectors from neutralizing antibody, and may allow repeated Ad vector administration. PMID- 10804283 TI - Dose- and sex-related carcinogenesis by N-bis(2-hydroxypropyl)nitrosamine in Wistar rats. AB - An initiation-promotion medium-term bioassay for detection of chemical carcinogens, developed in the male F344 rat, uses 0.1% N-bis(2 hydroxypropyl)nitrosamine (DHPN) among five genotoxic chemicals for the initiation of carcinogenesis in multiple organs. To establish this bioassay in the Wistar strain, the effects of two dose levels of DHPN were evaluated on the main DHPN rat target organs: lung, thyroid gland, kidneys and liver. Four groups of male and female animals were studied: Control -- untreated group; Multi-organ initiated group (also referred to as DMBDD, based on the initials of the five initiators) -- treated sequentially with N-diethylnitrosamine (DEN, i.p.), N methyl-N-nitrosourea (MNU, i.p.), N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN, drinking water), N, N'-dimethylhydrazine (DMH, s.c.) and DHPN (drinking water) for 4 weeks; a third group treated with 0.1% DHPN in drinking water for 2 weeks and the last group treated with 0.2% DHPN in drinking water for 4 weeks. The animals were sacrificed after 30 weeks. DHPN at 0. 2% induced preneoplasia in the liver and kidneys of rats of both sexes, the number and area of the putative preneoplastic liver glutathione S-transferase-positive hepatocyte foci being significantly increased in these animals. It also induced benign and malignant tumors in female and in male rats. However, there was no relationship between the increased incidence of preneoplastic lesions and tumor development in the 0.2% DHPN-exposed groups of both sexes. DHPN at 0.1% induced only a few preneoplastic lesions in the liver and kidney and no tumors in both male and female rats. A clear dose and sex-related carcinogenic activity of DHPN was registered, although Wistar rats of both sexes showed a relative resistance to the carcinogenic activity of this compound. PMID- 10804284 TI - Effects of reproduction on spontaneous development of endometrial adenocarcinomas and mammary tumors in Donryu rats. AB - Effects of reproduction on spontaneous development of uterine endometrial adenocarcinomas and mammary tumors in Donryu rats were investigated. While the incidence of endometrial adenocarcinomas in Donryu rats was not influenced by a single reproductive experience (SRE), it showed a tendency to decrease in animals having three reproductive experiences (TRE), compared to the nulliparous case (NRE). In addition, both SRE and TRE animals showed delayed occurrence and decreased incidences and mean numbers of mammary tumors, along with reduced incidences of proliferative lesions in the pituitary gland and mucinous epithelium in the vagina. The appearance-time and incidences of persistent estrus in TRE rats were delayed and low, respectively, compared to the SRE and NRE values. The hormonal environment was altered in both groups, the prolactin level in TRE especially being decreased. These results suggest that suppression of the occurrence of endometrial adenocarcinomas and mammary tumors in rats experiencing reproduction is associated with change in the hormonal milieu. PMID- 10804286 TI - Analysis of novel metastasis-associated gene TI-227. AB - TI-227 is a cancer metastasis-associated gene isolated from the B16-F10 mouse melanoma subline that preferentially metastasizes to the lung following intravenous injection. TI-227 is more highly expressed in the B16-F10 and colon26 nl17 sublines, which preferentially metastasize to the lung following intravenous injection, than in the B16-BL6 and colon26-nl22 sublines, which preferentially metastasize to the lung following subcutaneous injection. Since TI-227 could not be detected in normal tissues, there is a possibility that it is expressed specifically in tumors that preferentially metastasize to the lung following intravenous inoculation. Although the 5' end of TI-227 was analyzed, its total sequence contained no obvious open reading frame. However comparison with the isolated human counterpart, TI-227H gene, revealed a highly homologous region near the 3' end, which was expected to contain a very short open reading frame. PMID- 10804285 TI - Chinese medicinal herb, Acanthopanax gracilistylus, extract induces cell cycle arrest of human tumor cells in vitro. AB - We investigated the effect of a Chinese medicinal herb, Acanthopanax gracilistylus (AG), extract (E) on the growth of human tumor cell lines in vitro. AGE markedly inhibited the proliferation of several tumor cell lines such as MT 2, Raji, HL-60, TMK-1 and HSC-2. The activity was associated with a protein of 60 kDa, which was purified by gel-filtration chromatography. Cell viability analyses indicated that the treatment with AGE inhibits cell proliferation, but does not induce cell death. The mechanism of AGE-induced inhibition of tumor cell growth involves arrest of the cell cycle at the G(0) / G(1) stage without a direct cytotoxic effect. The cell cycle arrest induced by AGE was accompanied by a decrease of phosphorylated retinoblastoma (Rb) protein. Furthermore, cyclin dependent kinases 2 and 4 (Cdk2 and Cdk4), which are involved in the phosphorylation of Rb, were also decreased. These results suggest that AGE inhibits tumor cell growth by affecting phosphorylated Rb proteins and Cdks. PMID- 10804287 TI - Mutation of the SRC gene in endometrial carcinoma. AB - Recently, an activating mutation of the SRC gene has been implicated in about one tenth of advanced colon cancers. The SRC 531 mutation results in truncation of SRC directly C-terminal to the regulatory Tyr 530 and appears to activate the Tyr 530. To investigate whether mutation of SRC plays an important role in the development and progression of gynecological tumors, we performed mutational analysis of the entire coding region of SRC in 70 ovarian carcinomas, 68 endometrial carcinomas and 3 endometrial stromal sarcomas by means of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) followed by nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis. We found one truncated mutation at codon 531 (Gln to Stop) in an endometrial carcinoma. However, we found no mutation of this gene in ovarian carcinoma or endometrial stromal sarcoma. Our results suggest that mutation of SRC may be implicated in a small proportion of endometrial carcinomas. PMID- 10804288 TI - Screening of BRCA1 mutation using immunohistochemical staining with C-terminal and N-terminal antibodies in familial ovarian cancers. AB - We examined the subcellular localization of BRCA1 proteins using immunohistochemical staining with C-terminal (GLK-2 antibody) and N-terminal (Ab 2 antibody) monoclonal antibodies in 44 familial ovarian cancers. Among these, 24 cases were associated with 13 independent germ-line mutations of BRCA1, and loss of heterozygosity (LOH) at one or more BRCA1 microsatellite markers was found in all 21 informative tumors tested. With GLK-2 antibody, cytoplasmic staining was observed in 15 of 16 tumors (93.8%) with mutation in exon 11, and BRCA1 staining was absent in 8 of 8 tumors (100%) with mutation in exons other than exon 11. When immunohistochemical staining was performed with Ab-2 antibody, both nuclear and cytoplasmic staining were observed in 14 of 16 tumors (87.5%) with mutation in exon 11. Interestingly, nuclear staining was observed in 3 of 3 tumors (100%) with mutation downstream of exon 11, even though no staining was detected in 5 of 5 tumors (100%) with mutation upstream of exon 11. On the other hand, in familial ovarian cancers without BRCA1 mutations, nuclear staining or both nuclear and cytoplasmic staining was observed in 18 of 20 specimens (90%) and 20 of 20 specimens (100%) with GLK-2 antibody and with Ab-2 antibody, respectively. These results suggest that an immunohistochemical assay in combination with employing the C-terminal and the N-terminal antibodies appears to have potential as a reliable and useful technique for the screening of BRCA1 mutations, at least to predict the status of mutation, upstream or downstream of exon 11. PMID- 10804289 TI - Clinicopathological significance of expression of CD44 variants in head and neck squamous cell carcinoma. AB - Splice variants of the cell surface glycoprotein CD44 have been reported to be associated with the progression of various human tumors. The aim of this study is to determine the correlation between the expression of CD44 isoforms, especially CD44 variant 2 (CD44v2), and the clinicopathological features of head and neck squamous cell carcinomas (HNSCCs). The expression of CD44 isoforms was evaluated immunohistochemically in paraffin-embedded tissues from 89 primary lesions, using monoclonal antibodies against CD44 standard (CD44st), CD44 variant 6 (CD44v6) and CD44v2. Cancer tissues from 89 (100%), 85 (95.5%) and 59 (66.3%) patients showed positive immunoreactivity for CD44st, CD44v6 and CD44v2, respectively. A significant correlation was observed between the down-regulation of CD44v2 and poorer differentiation of the tumor cells (P = 0.02). We could not find any significant correlation between the expression of CD44v2 and T stage or N stage (lymph node status). However, the rate of positive cervical lymph node metastasis tended to increase with reduced expression of CD44v2 (P = 0.08). Down-regulation of CD44v2 expression was correlated with shorter overall survival (P = 0.01). Furthermore, Cox's multivariate analysis revealed that only CD44v2 expression and lymph node status were independent prognostic factors. These findings suggest that down-regulation of CD44v2 expression may be one of the biological markers for the degree of malignancy in HNSCCs. PMID- 10804290 TI - Negative p53/positive p21 immunostaining is a predictor of favorable response to chemotherapy in patients with locally advanced bladder cancer. AB - The relationship between clinical response to DNA-damaging drugs and p53 and p21 status in patients with locally advanced transitional cell carcinoma (TCC) of the bladder was assessed. The response to intraarterial chemotherapy (IAC) comprising 100 mg / m(2) of cisplatin (CDDP) and 40 mg / m(2) of pirarubicin (THP) and the prognosis were assessed in 23 patients (the mean follow-up period was 19 months). The p53 gene status of tumors was analyzed at exons 5 - 8 using polymerase chain reaction-single strand conformation polymorphism analysis in 19 patients, and paraffin-embedded tumor sections were immunostained for p53 and p21 in 23 patients. The overall objective response rate (incidence of good responders) was 70%. The negative p53 group (n = 17) showed a significantly higher objective response rate than the positive p53 group (n = 6) (82% vs. 33%; P = 0.045). The p53 gene status or p21 staining status was not significantly associated with responsiveness. When the p53 and p21 immunostaining results were combined, good responders were more accurately predicted than by p53 staining status alone; the negative p53 / positive p21 group (n = 12) showed an objective response rate of 92%, which was significantly higher than that of the positive p53 and / or negative p21 group (45%, n = 11) (P = 0.027). Cause-specific survival of the negative p53 group was significantly superior to that of the positive p53 group (P = 0.015). Negative p53 / positive p21 immunostaining is a possible predictor of favorable chemotherapeutic response in patients with TCC of the bladder. PMID- 10804291 TI - Schedule-dependent interactions between raltitrexed and cisplatin in human carcinoma cell lines in vitro. AB - Raltitrexed ('Tomudex') is a new anticancer agent which inhibits thymidylate synthase. To provide a rational basis for clinical trial design of the combination of raltitrexed and cisplatin, we studied the cytotoxic effects of this combination using various schedules in vitro and four human colon cancer cell lines, Colo201, Colo320, LoVo, and WiDr. Cell growth inhibition after 5 days was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of drug combinations at the concentration producing 80% cell growth inhibition (IC(80)) level were analyzed by the isobologram method. Simultaneous exposure to raltitrexed and cisplatin for 24 h, and sequential exposure to raltitrexed followed by cisplatin produced additive effects in the Colo201, Colo320, and LoVo cells, and additive and synergistic effects in WiDr cells. Sequential exposure to cisplatin followed by raltitrexed produced additive effects in the Colo201 cells and antagonistic effects in other three cell lines. Simultaneous and continuous exposure to both agents for 5 days produced additive effects in all four cell lines. These findings suggest that the simultaneous administration of raltitrexed and cisplatin, or the sequential administration of raltitrexed followed by cisplatin, generally produce the expected cytotoxicity at the cellular level and are optimal schedules, while the sequential administration of cisplatin followed by raltitrexed produces antagonistic effects and is inappropriate for this combination. Further in vivo and clinical studies will be necessary to determine the toxicity and antitumor effects of this schedule. PMID- 10804292 TI - A novel class of antitumor prodrug, 1-(2'-oxopropyl)-5-fluorouracil (OFU001), that releases 5-fluorouracil upon hypoxic irradiation. AB - We have been developing prodrugs of anticancer agents such as 5-fluorouracil (5 FU) that are activated by irradiation under hypoxic conditions via one-electron reduction. Among them, OFU001 [1-(2'-oxopropyl)-5-fluorouracil] is a prototype radiation-activated prodrug. In this study, we investigated the radiation chemical reactivity and the biological effects of OFU001. This prodrug is presumed to release 5-FU through incorporation of hydrated electrons into the antibonding sigma * orbital of the C(1')-N(1) bond. Hydrated electrons are active species derived from radiolysis of water, but are readily deactivated by O(2) into superoxide anion radicals (O(2).(-)) under conditions of aerobic irradiation. Therefore, 5-FU release occurs highly specifically upon irradiation under hypoxic conditions. OFU001 dissolved in phosphate buffer released 5-FU with a G-value (mol number of molecules that are decomposed or produced by 1 J of absorbed radiation energy) of 1.9 x 10(-7) mol / J following hypoxic irradiation, while the G-value for 5-FU release was 1.0 x 10(-8) mol/J following aerobic irradiation. However, the G-values for decomposition of OFU001 were almost the same, i.e., 3.4 x 10(-7) mol/J following hypoxic irradiation and 2. 5 x 10(-7) mol / J following aerobic irradiation. When hypoxically irradiated (7.5 - 30 Gy) OFU001 was added to murine SCCVII cells for 1 - 24 h, a significant cell-killing effect was observed. The degree of this cytotoxicity was consistent with that of authentic 5-FU at the corresponding concentrations. On the other hand, cytotoxicity was minimal when the cells were treated with aerobically irradiated or unirradiated OFU001. This compound had no radiosensitizing effect against SCCVII cells under either aerobic or hypoxic conditions when the drug was removed immediately after irradiation. Since hypoxia is generally most marked in tumors and irradiation is applied at the tumor site, this concept of prodrug design appears to be potentially useful for selective tumor treatment with minimal adverse effects of anticancer agents. PMID- 10804293 TI - Photodynamic inactivation with acridine orange on a multidrug-resistant mouse osteosarcoma cell line. AB - Overcoming multidrug resistance (MDR) is an urgent issue to improve the prognosis of osteosarcoma patients. In this study, we undertook to clarify the effect of photodynamic therapy (PDT) with acridine orange (AO) on the MDR mouse osteosarcoma (MOS / ADR1) cell line, by comparing the outcome with the effect on a chemosensitive osteosarcoma (MOS) cell line. Cultured cells of MOS and MOS / ADR1 cell lines were exposed to AO at various concentrations for various times, followed by long- or short-term (10 or 1 min) illumination with blue light (466.5 nm) for excitation. Living cells were counted by means of the trypan blue exclusion test. The results showed that AO rapidly bound to DNA, RNA and lysosomes of living MOS and MOS / ADR1 cells and also that most tumor cells in both cell lines died rapidly (viability ratio to untreated cells: 1/1000) within 48 h under conditions of continuous or 15-min flash exposure to AO at concentrations above 1.0 microg/ml plus 10-min illumination with blue light. Even after flash exposure to AO at concentrations above 1.0 microg/ml plus 1-min illumination, the viability of MOS/ADR1 cells decreased to a viability ratio of less than 1/ 1000 within 72 h. Based on these results, we concluded that AO with photo-excitation has a strong cytocidal effect, not only on chemosensitive mouse osteosarcoma cells, but also on MDR mouse osteosarcoma cells. These results suggested that photodynamic therapy with AO may be a new approach to treating MDR human osteosarcomas. PMID- 10804294 TI - Biodistribution of boron concentration on melanoma-bearing hamsters after administration of p-, m-, o-boronophenylalanine. AB - Although p-boronophenylalanine (p-BPA), a boronate analogue of tyrosine, has proven to be one of the most successful compounds for boron neutron capture therapy (BNCT) of malignant melanoma, the selective uptake mechanism of this compound into melanoma cells is not well understood. Therefore, the relationship between the structure of BPA and its specific affinity to melanoma cells appears worthy of investigation. In the present study, m- and o-boronophenylalanine (m- and o-BPA) were administered to melanoma-bearing hamsters and their uptake was measured. The time courses (0.5, 2.0, 4.0 and 48.0 h) of boron concentrations in melanoma, normal skin, and blood were determined in male Syrian (golden) hamsters bearing Greene's melanomas following a single intraperitoneal injection of either p-, m- or o-BPA (100 mg/kg of BPA fructose in 1.0 ml of saline). The boron concentrations in these tissues were measured by inductively coupled plasma atomic emission spectrometry (ICP-AES). In melanoma, the order of boron uptake was p- > m- > o-BPA at all time points, and the boron concentrations obtained with p-BPA and m-BPA resembled each other in that they had a peak at 2 h after administration and decreased with time. The melanoma/skin boron concentration ratio of p-BPA had a peak at 4 h after administration and the ratio ranged between 7/1 and 8/1. On the other hand, m-BPA and o-BPA had a peak at 2 h and their ratios ranged between 4/1 and 5/1. The difference in the accumulations of p BPA and m-BPA could be due to a difference in the property of p-BPA as a tyrosine analogue for melanin synthesis. The accumulation of m-BPA into melanoma might indicate the baseline level of metabolism-related amino acid transport. Our experimental findings indicate that this melanin synthesis, or the structural analogy between the boron compound and tyrosine as a precursor of melanin, is an important factor in the increased accumulation of p-BPA in melanoma cells. PMID- 10804295 TI - Evaluation of the interobserver agreement in the number of mitotic figures of breast carcinoma as simulation of quality monitoring in the Japan National Surgical Adjuvant Study of Breast Cancer (NSAS-BC) protocol. AB - In the National Surgical Adjuvant Study for Breast Cancer (NSAS-BC), node negative breast cancers were divided into higher- and lower-risk groups according to the histopathological nuclear grade given at individual collaborating hospitals, and the higher-risk group was entered into a randomized protocol of adjuvant therapy. Because the nuclear grade was the composite of nuclear atypia and mitotic counts, maintenance of interobserver agreement in mitotic counts was indispensable for the success of the protocol study. Fourteen pathologists participating in the protocol judged whether or not 20 photomicrographs suspected of showing mitotic cancer-cell figures truly showed mitoses. After standardizing the counting method, these pathologists counted the number of mitotic figures per 10 high-power fields of hematoxylin-eosin-stained main-tissue sections of 20 tumors. Areas where mitotic counts were considered to be the most frequent by each pathologist were compared for these tumors. For the judgment of whether the photomicrograph indicated mitosis, the level of interobserver agreement was moderate (kappa = 0.569). In the observations of 20 tumors, interobserver agreement level of mitotic counts was moderate (kappa = 0.506), that of nuclear atypia scoring was fair (kappa = 0.265), and that of nuclear grading was substantial (kappa = 0.633). The counted area was almost the same among the observers in 9 tumors, split into two areas in 6, and dispersed in 5. Concordance in judgment was achieved in 7 of the first 9 and in all of the third 5, but only in one of the second 6. The cause of discordance was mostly derived from tumor heterogeneity and the difference in the site where mitoses were counted. Interobserver agreement level was considered to be satisfactory, and it was expected that the case entry would be performed appropriately in the protocol study. The selection of the counting area was confirmed to be important for the acquisition of high-level agreement level in mitotic counts. PMID- 10804296 TI - [What's the true value of a consensus conference?]. PMID- 10804297 TI - [Sentinel node dissection in melanoma]. PMID- 10804298 TI - [Epidemiology tools in dermatology]. PMID- 10804299 TI - [Polyclonal T lymphocyte proliferation in drug lymphocyte activation test]. AB - BACKGROUND: We examined the clonality of the specific T cell reaction activated with specific drug antigens in patients with a positive lymphocyte activation test. PATIENTS AND METHODS: Ten patients were investigated. Their clinical features suggested drug allergy. Six weeks after a skin reaction, skin tests and lymphocyte activation tests were run. For each lymphocyte activation test, we explored the clonality of the T cell proliferation by examining the VJ rearrangement of the TCR gamma locus using polymerase chain reaction and denaturing gradient gel electrophoresis. RESULTS: All samples gave positive lymphocyte activation tests proving in vitro polyclonal T cell proliferation in reaction to a drug. There was no difference in clonality between the pilot lymphocyte activation test and the lymphocyte activation test after stimulation with phytohemagglutinin or with the drug antigen. DISCUSSION: Our findings show that in vitro T cell proliferation induced by a specific drug antigen as disclosed by the lymphocyte activation test is a polyclonal or oligoclonal cell proliferation. This lack of production of a preferential T clone in a T cell mediated immune drug reaction implies that many T lymphocytes can recognize the antigen and be stimulated by the same drug. PMID- 10804300 TI - [Descriptive epidemiological study of acne on scholar pupils in France during autumn 1996]. AB - BACKGROUND: Acne is the most common symptom prompting patients to consult a dermatologist. No previous study has been conducted in France to determine the prevalence of acne and describe the main epidemiological features. SUBJECTS AND METHODS: A cross sectional study was conducted in November 1996 and included 913 school children aged 11 to 18 years. This sample was statistically representative of the entire secondary school population in France during the 1996-1997 school year. The subjects were stratified by 5 criteria: age, sex, rural or urban residence, sun exposure, type of school. RESULTS: Taking the clinical diagnosis made by the dermatologist investigator as the main criteria, the overall prevalence of acne was 72 p. 100. It was 76.1 p. 100 using the new ECLA grading system previously described. The prevalence of acne was sex and age dependent: highest scores were found for girls aged 14-16 years and for boys aged 16-17 years. Genetic factors were very important for the outcome of acne. Finally, 41 p. 100 of the acneic subjects were following a treatment, prescribed by a dermatologist in two-third of the cases. DISCUSSION: These results are in agreement with those previously published in the literature although some differences were disclosed. It would appear important to distinguish between minimal acne with a few retentional pimples occuring during adolescence and severe acne (more than 20 pimples on the face) requiring early medical care to avoid scarring. PMID- 10804301 TI - [Oral terbinafine-induced plantar pustular psoriasis]. AB - BACKGROUND: Cutaneous reactions may occur in patients receiving terbinafine therapy, mainly rash and urticaria. More exceptionally, development of psoriasis has been described. We describe the development of plantar pustular psoriasis in a patient who took oral terbinafine. CASE REPORT: A 70-year-old man, without previously known history of psoriasis, was treated with terbinafine, 250 mg/day, for onychomycosis. Ten days later, a plantar pustular psoriasis appeared. The psoriasis cleared up after discontinuation of terbinafine and institution of antipsoriatic therapy. DISCUSSION: Terbinafine is an antifungal agent widely use in the treatment of onychomycosis. A skin reaction occurs in 3 p. 100 of patients. Only 24 cases of flare-up of previous psoriasis or psoriasis de novo after terbinafine have been described. The psoriasis generally clears after discontinuation of terbinafine. So, terbinafine is perhaps not the best first line therapy for onychomycosis in patients with psoriasis. PMID- 10804302 TI - [Vogt-Koyanagi-Harada's disease: 3 cases]. AB - BACKGROUND: Vogt-Koyanagi-Harada's disease is a bilateral chronic panuveitis sometimes associated with signs of meningo-encephalic, auditory and skin and nail involvement. We report 3 cases. CASE REPORTS: The first case was a 30-year-old woman who consulted for a red eye, low visual acuity, poliosis, and diffuse alopecia which had developed over 9 months. The ophthalmology examination disclosed anterior uveitis with retinal detachment. The second patient was a 9 year-old child who developed poliosis, canities, and achromic lesions over a 2 month period. The ophthalmology examination disclosed low visual acuity, irido corneal synechiae and pigmented deposits on the anterior lens. The third patient was a 20-year-old man who consulted for alopecia, diffuse canities, and white body hair. The ophthalmology examination disclosed low visual acuity, anterior uveitis, and a serous chorio-retinal detachment. All three patients were given general corticosteroid therapy (1 mg/kg/d). The clinical course was favorable in all cases with however one case of recurrent ocular involvement and one case of pigmentation disorders. DISCUSSION: The diagnosis of Vogt-Koyanagi-Harada's disease was established in these three cases on the basis of the ocular and skin and nail signs. This rare disease usually occurs in young, often female, patients. The pathogenesis remains unknown. Among the three signs observed, ocular involvement is the most serious. Skin and nail signs are seen in two thirds of cases. For rapid diagnosis and early treatment, this disease requires a multidisiplinary management associating the dermatologist and the ophthalmologist. PMID- 10804303 TI - [Congenital hypotrichosis and milia with spontaneous regression during adolescence or Oley syndrome: a variant of Bazex-Dupre-Christol syndrome]. AB - BACKGROUND: We report a family presenting the syndrome initially described by Oley characterized by congenital profus milia and hypotrichosis that regress during adolescence. CASE REPORT: A female infant with severe congenital hypotrichosis had profus milia involving the entire face. The remainder of the physical examination was normal. The mother had normal skin and hair but indicated she had had the same signs as a child. The patient's condition regressed from the age of 10 to 15 years but she has undergone several surgical resections for basocellular carcinoma since the age of 20. The maternal grandfather had spontaneously regressive typical follicular atrophodermia involving the back and the hands and also had several milium grains and several basocellular carcinomas. He had never presented hypotrichosis. DISCUSSION: Oley syndrome is defined as an association of congenital hypotrichosis and milia spontaneously regressive during adolescence. The symptoms presented by our patient and her mother are similar to this genodermatosis. Concomitant hypotrichosis, milia, basocellular carcinomas and follicular atrophodermia define the Bazex-Dupre-Christol syndrome. The grandfather's condition would be closer to this syndrome than Oley syndrome despite the spontaneous regression of certain anomalies during adolescence. This family study would suggest that the Bazex Dupre-Christol syndrome and the Oley syndrome are two variants of the same condition. PMID- 10804304 TI - [Papular mucinosis associated with myopathy]. AB - BACKGROUND: Papular mucinosis is an uncommon condition sometimes associated with myopathy as in our case. CASE REPORT: A 54-year-old woman presented several disseminated pruriginous papules that spread progressively. Histopathology of a skin biopsy showed mucine deposits and fibroblasts in the derma. Serum monoclonal lambda IgG was evidenced. The patient complained of weakness of the proximal muscles and dysphagia. Serum creatine phosphokinase was moderately elevated and the electromyography showed a typical myogenic pattern. Histopathology of a muscle biopsy disclosed a polymyositis aspect without mucine deposits. The patient was given prednisone. The skin lesions regressed totally with no modification of the muscle deficit. DISCUSSION: Myopathy associated with papulous mucinosis generally presents as a proximal deficit with dysphagia. Myalgia and amyotrophy are exceptional. Moderate serum myolysis is often evidenced. The electromyogram shows a myogenic pattern. The pathophysiology of this condition remains unclear, but the association with papulous mucinosis does not appear to be fortuitous due to its frequency. No standard treatment has been established. General corticosteroid therapy is generally effective and methotrexate, melphalan, isotretinoine and D-penicillamine have given good results in a few cases. PMID- 10804305 TI - [Facial hemangioma associated with arterial anomalies, coarctation of the aorta, and eye abnormalities: PHACES syndrome]. AB - BACKGROUND: Hemangiomas are frequent in childhood. Their association with dysmorphic anomalies is rare. Recently, the acronym "PHACES syndrome" was proposed to emphasize the association of Posterior fossa malformations, Hemangiomas, Arterial anomalies, Coarctation of the aorta and cardiac defects, Eye abnormalities, and Sternal malformations. CASE REPORT: A female child, 3 months old, had a large facial hemangioma. The physical examination was normal otherwise. A choroidal hemangioma and a papillary abnormality, causing amblyopia, were detected. The brain magnetic resonance imaging was normal. A subglottic hemangioma was found at endoscopy. At the age of 16 months, physical examination disclosed a heart murmur and coarctation of the aorta was detected. Moreover, the cardiac angiography showed diffuse arterial lesions. Strict surveillance was decided as there were no manifestations. DISCUSSION: Different abnormalities have been described to be associated with large facial hemangiomas. Frieden has grouped these abnormalities under the acronym PHACES. She described 43 hemangiomas and found 74 p. 100 Dandy Walker malformations and other posterior fossa malformations, 41 p. 100 arterial anomalies, 26 p. 100 cardiac or aortic malformations, 23 p. 100 ophthalmologic abnormalities. There is a high risk for the hemangiomas to develop in an airway localization. The prevalence of facial hemangiomas associated with other malformations is, to our knowledge, not known. In our department, 56 children were treated with corticosteroid therapy for severe facial hemangioma. 11 p. 100 had a cerebral abnormality. There were no cases with cardiac malformation or dysmorphism. PHACES syndrome is very rare but easy to remember. Thus in patients presenting a large facial hemangioma, it is important to conduct an attentive neurological examination completed by brain imaging and an extensive cardiovascular exploration. Special attention should be given to the ophthalmologic and sternal examinations as well as the search for hemangiomas in an airway localization. PMID- 10804306 TI - [Solar urticaria: pre-PUVA UVA desensitization]. AB - BACKGROUND: Solar urticaria is an uncommon disorder sometimes difficult to treat. It is characterized by the occurrence of typical whealing reactions on exposed skin a few minutes after sun exposure. The reactions resolve 1 to 5 hours after sun exposure ceases. We report a case evolving over several years, unresponsive to antihistamines and successfully treated by PUVAtherapy performed after UVA desensitization. CASE REPORT: For 3 years, a 22-year-old man developed erythema and itchy wheals at each solar exposure. The lesions appeared on all exposed areas including those usually exposed (face and hand) even in winter. An antihistamine regimen given for several weeks (cetirizine, loratadine) was ineffective. One trial of PUVA therapy led to an urticarial reaction of the entire body. Phototesting showed the minimal whealing dose for UVA was 0.4 J/cm(2). Phototherapy was therefore started by segmentary UVA irradiation at an initial dose of 0.1 J/cm(2). Exposure was then progressively increased allowing initiation of the PUVAtherapy on the 9(th) day with a dose of 0.5 J/cm(2) without whealing reaction. Slow increment PUVA therapy was able to induce good tolerance to sun exposure. DISCUSSION: Solar urticaria may sometimes have a deleterious effect on normal daily life. Severe cases are characterized by a whealing reaction after minimal sun exposure, even on regularly exposed skin. Antihistamines can provide some symptom relief in many patients, but high doses are required. If antihistamines are ineffective, PUVA therapy is indicated. Pre PUVA UVA desensitization is often necessary. However, exposure to UVA alone has to be repeated every 24 to 48 hours to maintain the refractory state. The advantage of PUVA therapy is a more long lasting protection allowing weekly maintenance sessions. PMID- 10804307 TI - [The future of histopathology of the skin. We disagree]. PMID- 10804308 TI - [Response to the anatomopathologists]. PMID- 10804309 TI - [Comments] [In Process Citation] PMID- 10804310 TI - [Giant cell arteritis revealed by scalp nodules]. PMID- 10804311 TI - [Pemphigus foliaceus on a 4-year-old female European cat]. PMID- 10804313 TI - [Dermatosis on above-knee and below-knee amputation stumps]. PMID- 10804312 TI - [Digital clubbing]. PMID- 10804314 TI - [Health economics, pharmacoeconomics and prescriptions in dermatology]. PMID- 10804315 TI - [Gene therapy for hereditary bullous epidermolysis]. PMID- 10804316 TI - [Giant cheloid scars]. PMID- 10804317 TI - [Brunauer-Fuhs striated palmoplantar keratoderma]. PMID- 10804318 TI - [Management of erysipela and necrotizing faciitis]. PMID- 10804319 TI - [Terbinafine-induced persistent agueusia: first reported case]. PMID- 10804320 TI - [University of Iowa Department of Dermatology]. PMID- 10804321 TI - [Consulting room at the hospital]. PMID- 10804322 TI - A need for therapeutic research in diabetic foot lesions healing. PMID- 10804323 TI - Non-alcoholic steatohepatitis: association with obesity and insulin resistance, and influence of weight loss. AB - Non-alcoholic steatohepatitis (NASH) is a disease of emerging identity and importance, and is now considered as one of the commonest liver diseases in western countries. It is frequently associated with severe obesity, especially abdominal adiposity, and is intimately related to various clinical and biological markers of the insulin resistance syndrome. Especially, both the prevalence and the severity of liver steatosis are related to male sex, body mass index, waist circumference, hyperinsulinaemia, hypertriglyceridaemia and impaired glucose tolerance or type 2 diabetes. A substantial weight loss following gastroplasty is accompanied by a marked reduction in the prevalence and the severity of the various biological abnormalities of the metabolic syndrome and, concomitantly, by an important regression of liver steatosis in most obese patients. However, in some patients, this rapid and drastic weight loss may result in a mild increase in inflammatory lesions (hepatitis), despite the regression of steatosis, which might result from the rapid mobilization of fatty acids or cytokines from adipose tissue, especially visceral fat. The intimate relationship between NASH and obesity leads to the concept that NASH may be considered as another disease of affluence, as is the insulin resistance syndrome and perhaps being part of it. PMID- 10804324 TI - Determination of lipoprotein(a) concentrations and apolipoprotein(a) molecular weights in diabetic patients. AB - Lipoprotein(a) (Lp(a)) with atherogenic and thrombotic properties has been frequently studied in diabetes, because a high cardiovascular risk has been reported both in type 1 and type 2 diabetes. Few studies have considered genetic factors, especially the isoforms of apolipoprotein(a). The aim of this work is to determine the distribution of apo(a) phenotypes in the serum of 148 diabetic patients (59 type 1, 89 type 2) with or without vascular complications. Apo(a) phenotypes are determined using 4-15% sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by immunoblotting (PhastSystem - Pharmacia). An inverse relationship is observed between Lp(a) serum concentration and the apparent molecular mass of apo(a) isoforms: type 1 r=- 0.61, p<0.01; type 2 r=- 0.55, p<0.01. The frequency of apo(a) isoforms is significantly different between type 1 and type 2 diabetes mellitus. A higher prevalence of isoforms of low molecular weight was observed in the type 2 diabetic population. PMID- 10804325 TI - Diabetic foot lesions: etiologic and prognostic factors. AB - The clinical characteristics of 132 diabetic patients referred for treatment of foot lesions were surveyed. One hundred and sixty three lesions (n=163) concerned 88 men and 44 women during a five-year period (from January 1989 to December 1993). Hospitalisation rate equalled 9.16%, i.e. 11.17% for men and 6.82% for women (p <0.001); the men/women ratio was 1.64. Eighty nine per cent (89%) of patients presented type 2 diabetes and 11% of patients type 1 diabetes. Mean age at the first foot lesion was 59.64 +/- 11.74 years. The mean duration of diabetes was 10.95 +/- 6.80 years. The patients had a high prevalence of diabetic complications, particularly peripheral neuropathy (84.85%) and obvious peripheral arteriopathy (78.78%). Infection was almost constant. There was no significant difference between men and women as far as the prevalence of complications was concerned. Smoking habits were noticed only in men. Inadequate footwear was considered as the major exogenous risk factor leading to a foot lesion. The definitive results 6 months after hospitalisation were as follows: the death rate was 9.09% (n=2; 11 men and 1 women, NS); 15.90% of patients (n=12) underwent a major amputation (4 at the level of the thigh, 17 at the level of the leg), 14.39% of patients (n=19) underwent a minor amputation; in 59.09% of patients (n=78) there was no amputation. Two patients (1.51%) underwent two consecutive amputations, left hospital against medical advice during their second hospitalisation, and then were lost sight. The prevalence of foot lesions was more important in men. Moreover, seriousness of the lesions and consequently the rate of amputations were important in men; this was probably due to smoking habits. The factors that influence the outcome seem to be: male gender, delay of management, quality of medical treatment, surgical attitude, inadequate level of amputation and finally lack of structured prevention. Prevention then should be based on the patient's education, general practitioners' training and a better and more efficient cooperation between surgeons and diabetologists. PMID- 10804326 TI - Continuous intraperitoneal insulin infusion partly restores the glucagon response to hypoglycaemia in type 1 diabetic patients. AB - The glycaemic and hormonal responses to a hypoglycaemic event induced by an i.v. bolus of insulin was studied in seven type 1 diabetic patients treated first with continuous subcutaneous insulin infusion (CSII) and subsequently with continuous intraperitoneal insulin infusion (CIPII). Arterialised blood glucose and venous hormonal responses were analyzed. HbA1c was improved by CIPII. Although a regimen of a higher basal insulin infusion rate was applied during CIPII the basal peripheral venous insulin levels were lower. The i.v. bolus of insulin resulted in hypoglycaemia in both tests but was more pronounced during the CSII test expressed as a smaller area under the curve (AUC) for the first hour (13.0 +/- 2.3 vs. 13.7 +/- 1.2 mmol l(-1) h(-1), p=0.016, CSII vs. CIPII). The hypoglycaemia resulted in a significant and similar increase in the plasma levels of adrenaline, cortisol and growth hormone in both experiments. A significant increase in the glucagon level was only observed during CIPII. The incremental glucagon response was also significantly more pronounced in the CIPII test expressed as maximal responses (7.5 +/- 3.0 vs. 17.0 +/- 3.1 pg ml(-1), p =0.048, CSII vs. CIPII) as well as incremental AUC (5.1 +/- 12.0 vs. 44.4 +/- 13.2 pg ml( 1) h(-1), p =0.027, CSII vs. CIPII). It seems that CIPII in type 1 diabetic patients could improve the glucagon release to hypoglycaemia. This observation may contribute in explaining why CIPII is associated with a lower incidence of hypoglycaemia in spite of an improvement in metabolic control. PMID- 10804327 TI - Cost-effectiveness modeling of Dermagraft for the treatment of diabetic foot ulcers in the french context. AB - To assess the cost-effectiveness of Dermagraft(R) (human dermal replacement) in the treatment of the diabetic foot ulcer, compared to standard treatment. A Markov model was developed, to simulate, over a 52-week period, the health status of a cohort of 100 patients with a diabetic foot ulcer treated either with conventional therapy or with Dermagraft(R). The considered health states were: healed, same site recurrence, unhealed not infected, cellulitis, osteomyelitis, amputation and death. Each week, the patient may progress among states according to a set of transition probabilities directly derived from the original clinical trial conducted in the USA. The cost of each health state was estimated by a Delphi panel of French diabetologists (direct costs only, valuated from a societal perspective). A sensitivity analysis was performed. The total number of healed ulcers included first ulcers healed (76.38% for Dermagraft(R) vs. 69.35% for standard treatment; median time to heal is 14-15 weeks for Dermagraft(R) compared with 28-29 weeks for standard treatment) plus recurrences which are subsequently healed within the 52-week period (14.29 for Dermagraft(R) vs. 25.09 for standard treatment; median time to heal is 3-4 weeks for Dermagraft(R) compared with 5-6 weeks for standard treatment). The average expected cost per treated patient (C/E) using standard treatment for the considered 52-week period is 47,418 FF vs. 54,384 FF for Dermagraft(R) (including 18,200 FF for Dermagraft(R) acquisition and 36,184 FF for standard treatment). Because Dermagraft(R) heals more ulcers within 52 weeks, the average cost per healed ulcer is lower (53,522 FF vs. 56,687 FF for standard treatment). The incremental cost-effectiveness ratio of Dermagraft(R) (DeltaC/DeltaE) equals 38,784 FF, indicating the extra investment that the decision-maker has to accept for an additional ulcer healed with Dermagraft(R) compared with conventional treatment. PMID- 10804328 TI - Lack of effect of a physiological elevation of plasma non-esterified fatty acid levels on insulin secretion. AB - Elevated plasma non-esterified fatty acid (NEFA) levels in obese subjects may contribute to their higher insulin secretory rates by direct effects on the islet B-cells. This may involve short-term metabolic effects, or long-term effects on islet B-cell mass, which is characteristically increased in obesity. We examined the effects of elevating plasma NEFA levels for 5.5 to 7 h on insulin secretion after an overnight fast and during a 90 min 12 mmol/l hyperglycemic clamp in 9 normal women (40.1 +/- 9.5 years [mean +/- SD]; BMI: 25.2 +/- 3.72 kg/m(2) ). Subjects were studied twice. In one study plasma NEFA levels were increased approximately 2-fold by infusion of 20% Intralipid (60 ml/h) and heparin (900 U/h) for 5.5 h before and throughout the glucose clamp. Elevated NEFA levels were associated with a small increase in fasting plasma glucose (5.0 +/- 0.1 vs 4.7 +/ 0.1 mmol/l, P <0.05) and C-peptide levels (0.54 +/- 0.09 vs 0.41 +/- 0.06 nmol/l, P <0.05). The increase in fasting insulin levels did not, however, reach statistical significance (9.0 +/- 2.5 vs 5.3 +/- 1.4 mU/l, NS). During the glucose clamp, plasma NEFA levels were suppressed to very low levels in the saline control study. Although plasma NEFA levels also fell in the lipid/heparin study, they remained significantly higher than on the control day, and somewhat higher than might be expected postprandially in obese subjects. During the glucose clamps, plasma glucose, insulin, and C-peptide profiles were similar on the two study days. No difference in either first or second phase insulin secretion was observed between the two studies. In conclusion, our findings do not support the idea that the exaggerated insulin secretion in obesity is mediated by short-term effects of plasma NEFA levels on islet B-cell metabolism, independent of plasma glucose levels. PMID- 10804329 TI - Effect of sulfur dioxide inhalation on erythrocyte antioxidant status and lipid peroxidation in experimental diabetes. AB - The effect of sulfur dioxide (SO(2) ) on red cell antioxidant status and lipid peroxidation was examined in this research. Forty healthy male albino rats, aged three months, were divided into four equal groups: Control (C), SO(2) +C (CSO(2) ), diabetic (D) and SO(2) +D (DSO(2) ). Experimental diabetes mellitus was induced by i.v injection of alloxan with a dose of 50 mg/kg body weight. Ten ppm SO(2) was administered to the animals of SO(2) exposed groups in an exposure chamber for one hr/day x 7 days/wk x 6wks while other groups were exposed to filtered air in the same condition. SO(2) exposure, while markedly decreasing Cu, Zn-Superoxide dismutase (Cu, Zn-SOD) activity, significantly increased glutathione peroxidase (GSH-Px), catalase (CAT), glutathione (GSH) and glutathione-s-transferase (GST) activities and TBARS values in CSO(2) and DSO(2) groups compared with their respective control groups. From these results, it could be concluded that adaptative changes occurred in antioxidant systems that may counteract the free radical effect of SO(2) in the experimental groups. PMID- 10804330 TI - Hypothermia with acute renal failure in a patient suffering from diabetic nephropathy and malnutrition. AB - We report a rare case of hypothermia with acute renal failure in a patient suffering from diabetic nephropathy. A 71-year-old male who had been receiving insulin therapy for the treatment of diabetes mellitus complicated with advanced diabetic nephropathy since 1998 was malnourished with an extremely decreased muscle mass. Without any prolonged exposure to excessively low external temperatures or hypothyroidism, pituitary insufficiency, adrenal insufficiency, sepsis, hypoglycemia, and diabetic ketoacidosis, acute hypothermia appeared together with an aggravation of diabetic nephropathy. His skin temperature fell to below measurable levels and his rectal temperature fell to 30.0 degrees C. His consciousness was drowsy and the hypothermia was not accompanied by shivering. Skeletal muscle is known to play an important role as a center of heat production and shivering thermogenesis in skeletal muscle mainly operates on acute cold stress. Therefore, in this case, hypothermia may have occurred because the shivering thermogenesis could not fully act on the acute cold stress due to the dramatically reduced muscle mass. We should always keep in mind that older, malnourished diabetic patients can easily suffer from impairments of the thermoregulatory system. PMID- 10804331 TI - [How to measure glycemic instability?]. AB - Instability of glycemic levels is "normal" in type 1 diabetes, with different levels of severity, up to the restrictive definition of brittle diabetes (repeated ketoacidosis and/or severe hypoglycemias). Quantification of glycemic instability, in terms of intraday variability and day-to-day reproducibility, is advisable. Standard deviation of blood glucose, though a simple index does not discriminate between slow and brutal variations. Repartition of blood glucose values also only indicates dispersion. M values compares the patient values to an ideal blood glucose level, emphasizing the role of low values. The MAGE index measures the amplitude of the largest glucose excursions, thus evaluating appropriately glycemic variability. The Low Blood Glucose Index (LBGI) is a new index of variability emphasising (as the M value) the low glycemias. Each glycemia is given a value from O (if >=110 mg/dl) to 100 (if 20 mg/dl). It thus integrates the frequency and severity of hypoglycemias. According to its authors the LBGI would be the best indicator of severe hypoglycemias. The Mean of Daily Differences (MODD) evaluates the day-to-day reproducibility of blood glucose values. All the above indexes could easily be incorporated in the programmes of large memory glucose meters. PMID- 10804332 TI - [Digestive tract endoscopy: the right sedation?]. PMID- 10804334 TI - [Patient opinions after colonoscopy. Results of a cross sectional, multicenter ANGH study (2)]. AB - OBJECTIVES: The aims of this study were to evaluate patient opinions following colonoscopy and to analyze the role of conditions of practice on patients' opinions. METHODS: A study was suggested to all members of the ANGH. During one week of practice, a questionnaire was filled out by the gastroenterologist and a second one by the patient, alone, one day after colonoscopy. RESULTS: Eighty six hospitals (229 gastroenterologists) took part in the study which included 815 patients; 752 of them (92%) filled out the satisfaction questionnaire. Five hundred and eighty four patients (78%) would accept a new endoscopy in the same conditions; 168 patients (22%) would accept if the conditions were different or would refuse a new endoscopy. The main factors associated with acceptance under the same conditions were: good tolerance (odds-ratio (OR): 5.08), old age (OR: 5), anesthesia (OR: 2.67) or conscious sedation (OR: 2.29), male gender (OR: 2.39) and ambulatory care (OR: 2). CONCLUSION: This study of clinical practice underscored the different factors associated with better acceptance of colonoscopy. Individual gastroenterologists could use these data to guide practice. PMID- 10804333 TI - [Conditions of practice and results of colonoscopy in non-university hospitals. Results of a cross sectional, multicenter ANGH study (1)]. AB - OBJECTIVES: The aim of the study was to describe the conditions of practice and results of colonoscopies performed in non university hospital endoscopy units. METHODS: A study was suggested to all members of the National Association of non university Hospital Gastroenterologists (ANGH). During one week a questionnaire was filled out by the gastroenterologists for each colonoscopy. RESULTS: Eighty six hospitals (229 gastroenterologists) took part in this study including 840 explorations. Ninety one percent of them were complete and the result was abnormal in 49%: cancers (3%), polyps > or =1 cm (8%), polyps < 1 cm (20%) and colitis (5%). Colonoscopies were ambulatory in half of the cases. Sedation was used in 95% of the patients, 80% by the anesthesist and 15% by the gastroenterologist (conscious sedation). The recommendations of the French Society of Anesthesia and Intensive Care for ambulatory sedation were not totally respected (15% of patients left the endoscopy unit without assistance). Only 6% of colonoscopies were not well tolerated, because of pain or incidents rarely necessitating intensive care monitoring (5 cases) or prolonged hospitalization (2 cases). CONCLUSION: Colonoscopy provides good diagnostic yield although conditions of use can be improved. PMID- 10804335 TI - [Colonic motility in humans. Recent physiological, pathophysiological and pharmacological data]. PMID- 10804337 TI - [Pre-test]. PMID- 10804336 TI - [Anal incontinence in adults. Guidelines for clinical practice. National French Gastroenterology Society]. PMID- 10804338 TI - [Synopsis: diagnosis and treatment of occult digestive tract bleeding]. PMID- 10804339 TI - [Comments on case report: A chronic, occult and obscure anemia of digestive origin]. PMID- 10804341 TI - [Answers to the pre-test]. PMID- 10804340 TI - [Interview: Questions to Pr. J. Boyer]. PMID- 10804342 TI - [Spontaneous peritonitis: a new indication for intravenous albumin?]. PMID- 10804343 TI - [Epidemiological information obtained from anti-hepatitis C virus screening in blood donors and candidates for autologous transfusion from 1992 to 1996 in the Alpes-Maritimes region]. AB - OBJECTIVES: To determine the evolution of the frequency of anti-hepatitis C virus antibodies from 1992 to 1996 in blood donors and in candidates for autologous transfusion in the Alpes-Maritimes region and to assess risk factors. METHODS: Anti-hepatitis C virus antibodies were assessed by second generation ELISA in 1992 and in the first quarter of 1993, and then by third generation ELISA; in all cases, anti-hepatitis C virus antibodies were confirmed by RIBA test. RESULTS: Since 1992 (when the second generation ELISA test became available), the prevalence of anti-hepatitis C virus antibodies in blood donors in the Alpes Maritimes region (0.54% in 1992 to 0.20% in 1996) has decreased. Positive ELISA anti-hepatitis C virus was confirmed by RIBA in 53 to 68% of anti-hepatitis C virus blood donations. The percentage of anti-hepatitis C virus donors with ALT above the upper limit (donation exclusion threshold) was between 28 and 56%.The most frequent age interval for new anti-hepatitis C virus positive donors was between 30 and 40 years. Since 1992, a third of the anti-hepatitis C virus blood donors agreed to participate in a medical history questionnaire. One or several risk factors were found in almost all donors. The most frequent source of infection was nosocomial (50%). During the 5 years of the study, the number of candidates for autologous transfusion increased: 717 in 1992 to 1 528 in 1996. The prevalence of anti-hepatitis C virus in this older population (mean age: 64 years) decreased progressively (2.9% in 1992 to 1.1% in 1996, P<0, 01) since the prevalence of anti-HBc remained stable, near 12%. Among the 96 subjects found to be anti-hepatitis C virus positive before an autologous transfusion, 49 were transfused before 1990 and 40 had a history of surgery. CONCLUSION: The prevalence of anti-hepatitis C virus has decreased since 1992 in blood donors and in candidates for autologous transfusion which may suggest that there is better screening in the general population and presenting the spread of hepatitis C virus infection. PMID- 10804344 TI - [Viability and differentiation of human hepatocytes immunoprotected by macroencapsulation and transplanted in rats]. AB - OBJECTIVES: To determine the viability and differentiation of human hepatocytes immunoprotected by encapsulation and transplanted in rats without immunosuppression. METHODS: Freshly isolated human hepatocytes were encapsulated in hollow fibers and transplanted in the peritoneal cavity of immunocompetent rats. The fibers were explanted for analysis at D3, D7 and D14 following transplantation. Morphological features under light and electron microscopy and gene expression were compared to those of non-transplanted encapsulated hepatocytes (D0). Human cytochrome P450 3A and albumin mRNAs were quantified by Northern blot. Cytochrome P450 3A proteins were detected by Western blot and cytochrome P450 3A enzyme activity was assessed by measuring the formation of 6beta-hydroxytestosterone by high performance liquid chromatography. RESULTS: Transplanted hepatocytes were more than 60 % viable and exhibited morphological criteria of hepatocytic differentiation up to D7. Albumin and cytochrome P450 3A transcripts were also detected up to D14. At D3 and D7, albumin mRNA levels were of 30 %, compared to control D0 hepatocytes, while cytochrome P450 3A5 and cytochrome P450 3A4 mRNA levels were 65 % and 0 %, respectively. Cytochrome P450 3A immunoreactivity was detected by Western blot up to D14 and 6beta-hydroxylase activity was 17 % at D3 compared to D0, supporting with disappearance of cytochrome P450 3A4 mRNA. CONCLUSIONS: Human hepatocytes remain viable for a short period, following encapsulation and intraperitoneal transplantation in rat. Other experimental conditions need to be tested to prevent or delay a decrease in hepatocyte specific gene expression. PMID- 10804345 TI - [Ageing, alcohol and mitochondria]. PMID- 10804346 TI - [Intra-abdominal desmoplastic small round-cell tumors. An entity among peritoneal carcinomatoses in young adults]. AB - Desmoplastic small round-cell tumors in young adults were first described in 1989. They may be revealed by an abdominal mass or a peritoneal carcinomatosis. The diagnosis and the initial treatment are based on debulking surgery. Chemotherapy using doxorubicin, ifosfamide, etoposide and cisplatin can then be proposed. Despite some initial chemosensitivity, prognosis remains poor. PMID- 10804347 TI - [Small bowel pseudo-obstruction revealing an early scleroderma. Long-term efficacy of octreotide and erythromycin]. AB - Skin abnormalities are the most common manifestations in patients with systemic sclerosis but many other organs can be involved. We report here the case of a 61 year-old-man in whom small bowel involvement was the initial presentation of limited systemic sclerosis. The clinical features consisted of recurrent small bowel obstruction without any organic lesion. The patient responded well to erythromycin (125 mg thrice a day before meals) and octreotide (50 microg subcutaneously at bedtime) with complete symptom relief. This permitted recovery of nutritional autonomy for two years. This observation shows that, in patients with intestinal pseudo-obstruction, systemic sclerosis should be considered. The long-term administration of octreotide and erythromycin can be effective in treating small bowel dysmotility in such patients. PMID- 10804348 TI - [Systemic sarcoidosis during interferon-alpha therapy for chronic hepatitis C virus infection]. AB - A 62-year-old white woman with chronic hepatitis C virus infection was diagnosed with systemic sarcoidosis during treatment with interferon-alpha. Seventeen cases of sarcoidosis associated with interferon treatment have been reported, with only 8 inpatients with chronic hepatitis C virus infection. Our patient, with no past history of sarcoidosis, developed subcutaneous sarcoid nodules, arthritis, uveitis, pneumonitis and heart involvement two months after she was started on treatment with interferon-alpha. Her symptoms resolved when she began corticosteroid therapy and interferon-alpha was stopped. This case report suggests that exogenous interferon-alpha may trigger or contribute to the development of this multisystem granulomatous disorder in which interferon-gamma and CD4 + T-lymphocyte are mainly involved. Patients should be monitored during and following interferon-alpha therapy, since the autoimmune diseases induced by interferon therapy do not always improve after cessation of treatment. PMID- 10804349 TI - [Biochemical and tumoral markers in a case of epidermoid cyst of an accessory spleen: be careful of the interpretation!]. PMID- 10804350 TI - [Colonic epithelial lymphocytosis associated with self-limited chronic Brainerd type diarrhea]. PMID- 10804351 TI - [A rare cause of acute abdominal pain: mesenteric heterotopic pancreas complicated by acute pancreatitis]. PMID- 10804353 TI - [Is xenical hepatotoxic?]. PMID- 10804352 TI - [Massive gastrointestinal bleeding due to duodenal variceal rupture: value of portal, transhepatic embolization]. PMID- 10804355 TI - [ [In Process Citation] PMID- 10804354 TI - [Hepatitis C and parenteral use of doping substances: an unknown route of transmission?]. PMID- 10804356 TI - [Monochorionic twin pregnancies : dangerous liasons]. AB - Monochorionic twin pregnancies are characterized by a shared placental circulation; this induces a high degree of hemodynamic interdependency between the twins. This is particularly dramatic in twin-to-twin transfusion syndrome or when one twin dies in utero. In both conditions, the severity of the hemodynamic imbalance can lead to significant sequelae in 15-25% of the cases. Ischemic accidents can be prevented by interruption of placental anastomoses through laser coagulation of their chorionic branches. This can be performed when intrauterine death of one twin is inevitable or as a first line treatment in severe TTS. In the latter indication it is important to compare this approach with serial amniodrainage. PMID- 10804357 TI - [Inherited thrombophilia and pregnancy]. AB - Inherited thrombophilia include deficiences of antithrombin III, protein C and protein S, and the factor V Leiden mutation, the prothrombin gene variant, and homozygosity for the thermolabile variant of methylenetetrahydrofolate reductase (MTHFR). The incidence of thromboembolism events during pregnancy and postpartum period among women with thrombophilia is not well known and depends on the prethrombotic state resulting from the interaction of the underlying thrombophilic defect(s), history of congenital thrombophilia, and additional risk factors. In that way, many patients with congenital thrombophilia will require antenatal thromboprophylaxis, the timing of which will depend on the patient's history and thrombophilic disorders. Low molecular weight heparin appeared to be a safe alternative to unfractionated heparin for both the fetus and the mother during the pregnancy. Case-control studies have recently demonstrated that serious obstetrical complications i. e severe preeclampsia, abruptio placentae, intrauterine growth restriction, and stillbirth were frequently associated with inherited thrombophilia. Controlled trials are now urgently needed to determine the possible potential benefits of anticoagulant therapy in pregnancy outcome. Finally, there is no evidence to support routine screening for congenital thrombophilia during pregnancy. PMID- 10804358 TI - [Severe precocious pre-eclampsia : how to manage the feto-maternal conflict of interest]. AB - BACKGROUND: The HELLP syndrome is usually treated by rapid termination of pregnancy as the most effective way of limiting the risk of disease aggravation and maternal complications. The drawback is the risk of fetal complications due to prematurity when pregnancy is terminated before 32 weeks gestation. A controlled study has suggested that corticosteroid therapy could be effective in early HELLP syndrome. PATIENTS AND METHODS: From January 1, 1996 to March 1, 1999, we treated patients presenting early HELLP syndrome defined as platelet counts below 150,000 and ALAT above 50 U/l prior to 32 weeks gestation with dexamethazone (Soludecadron((R))) via intravenous administration of 10 every 12 hours to the end of pregnancy or until platelet counts rose above 150,000. RESULTS: Among 14 patients with early HELLP syndrome, 10 were treated (including 5 primiparous women). Six patients among the 10 had platelet counts between 50,000 and 100,000. Mean term at the first injection of dexamethazone was 29 weeks 3 days. Platelet counts rose and transaminase levels fell in all patients. Pregnancy was prolonged 7 2 days. The only maternal complication was one case of disseminated intravascular coagulation. There were no neonatal deaths. DISCUSSION: Our results are similar to those reported in the literature but such a treatment scheme can only be instituted within a rigorously controlled monitoring system in a unit with neonatal and maternal intensive care facilities. This protocol remains safe for patients with HELLP syndrome whose platelet count remains at least above 50,000. PMID- 10804359 TI - [Bilateral agreements to reduce maternal mortality in developing countries. The FIGO experience]. AB - In 1997, the International Federation of Gynecology and Obstetrics (FIGO) launched its Save the Mother program. Its overall objective is to mobilise obstetricians and gynaecologists in developing and developed countries to work together in an effort to reduce maternal mortality, through operational research demonstrating the feasibility and effectiveness of integrated comprehensive essential obstetric services. The FIGO Save the Mothers Uganda - Canada Project, jointly led by the Society of Obstetricians and Gynaecologists of Canada and the Association of Obstetricians and Gynaecologists of Uganda (AOGU), is one of the projects supported by FIGO. Conducted in Kiboga (Uganda), the project aims the reduction of maternal mortality and morbidity by a series of measures which target the district hospitals, community dispensaries and the community. PMID- 10804360 TI - [Static and dynamic MRI of the normal and pathological female pelvic floor]. AB - Static and dynamic MRI of the female pelvic floor is a new application of this technology. The capability of MRI to assess cavities and muscles on the same exploration is an important improvement. The authors describe successively: MRI is nowadays a decisive tool in the preoperative work-up of pelvic floor dysfunctions. PMID- 10804361 TI - [Use of "TVT" in surgery for female urinary incontinence]. AB - The TVT procedure (tension-free vaginal tape) was described by U. Ulmsten (Sweden) in 1996. This new procedures questions the numerous procedures proposed for female stress urinary incontinence (SUI). The mesh is placed under the urethra (and not under the bladder neck) and cures SUI without important voiding difficulties. Use of this operation is growing fast in Europe and Australia and is beginning to be used in the USA. TTV is a minimally invasive technique using local or spinal anesthesia applicable as an ambulatory procedure. Peroperative complications are limited to bladder perforation (without detrimental consequences) and bleeding. Postoperative complications are very rare. Local tolerance of the prolene mesh is excellent. Failure rates have been very low, about 6% for SUI and more controversial for unstable bladder. Since January 1997, we have performed about 400 TVT either as single procedures or in association with prolapsus cure. After 1 to 3 years follow-up, cure rate in the first 156 SUI patients was 89.1%, almost the same with or without associated procedures. For the 36 patients with an urethral closure pressure less than 30 cm H(2)O, cure rate was 75%. In cases of mixed incontinence, the cure rate varied from 57.9% (associated procedures to 61.3% (single procedures). The rate of de novo instability was less than 4.6%. Induced and increased voiding difficulties (5.7% and 6% respectively) remained at a relatively low level provided the strict operative protocol was used. The TVT procedure is easy to perform, relatively harmless, reproducible and very efficient even for patients with risk factors, sphincteric weakness, previous operations or an associated prolapsus cure procedure. PMID- 10804362 TI - [Validation of laparoscopic surgery in gynecology]. AB - Laparoscopic surgery is presently the gold standard for treatment of benign adnexal pathologies although the evaluation of other surgical techniques such as laparoscopic assisted hysterectomy or myomectomy remains to be made. Multicenter observational studies and data collection should be pursued to evaluate these new surgical techniques as they spread from a few promoting centers. PMID- 10804363 TI - [Sentinel lymph node detection in breast cancer. Experience of the Institut Curie]. AB - Sentinel lymph node biopsy is a recently developed, minimally invasive technique for staging the axilla in breast cancer. This new procedure of selective lymphadenectomy has been the subject of several studies, and a consensus of opinion is starting to form to define indications and methods of identification concerning the use of this technique. At the Institut Curie since 1996, we have been using the Patente blue dye technique and from 1998 we have used the combination of blue dye and technetium labeled sulfur colloid. This article summarizes the principales aspect of this technique. PMID- 10804364 TI - [New therapeutic plans for cancer of the cervix]. AB - New methods of staging (MRI, laparoscopic surgery) and therapy (concomitant radio chemotherapy, debulking of diseased nodes), with a trend towards the reduction of the aggressivity of surgical procedures (minimal access surgery, ovarian preservation, reduction of the extent of radical hysterectomy, uterine conservation) may and should by integrated in new therapeutic protocols. As a consequence, the management of invasive cervical cancer becomes even more multidisciplinary and specialized. PMID- 10804365 TI - [Vaccination against human papillomavirus and against the cellular proliferation that it induces: facts and expectations]. AB - Vaccination against human papillomavirus (HPV) give promising results in animal models. Prophylactic vaccination uses essentially virus-like particles autoassembled from the L1 capsid protein in its native form. Protection is achieved: it is efficient, type-specific and humoral. Therapeutic vaccination is of cellular origin and raised against the E7 oncoprotein. Several vaccine preparations have been found to be efficient: they include virus-like particles formed with L1-E7 fusion protein, E7 encoding plasmidic DNA, dendritic cells pulsed with E7-derived peptides,. PMID- 10804366 TI - [Supraconservative breast surgery]. AB - Currently, 30% of the breast biopsies are performed for patients with nonpalpable lesions. The surgical management of these lesions had to evolve to a better three dimensional targeting and a reduction of the tissue traumatism. The ABBI procedure allows the percutaneous one bloc excision of suspicious mammographically detected lesions with a diameter of less than 2cm. We prospectively evaluated this procedure as a therapeutic tool. Of the 10 malignant lesions with a pathologic size <10mm, 9 (90%) were completely resected with the ABBI device (no residual disease at re-excision of the biopsy site). The results of this preliminary study suggest a potential therapeutic role of the ABBI procedure in the therapeutic arsenal against mammary lesions. PMID- 10804367 TI - [Fetal echocardiography volume mode (3 dimensions)]. AB - About 10 out of 100 newborns are affected by congenital heart defects that remain the least easy to detect remain prenatally, despite the relatively high detection rate of fetal malformations. Antenatal diagnosis of congenital heart defects depends on operator experience and skill, but recent techniques of computer assisted ultrasound imaging should be useful by allowing easier recognition of fetal heart anomalies. New imaging procedures include a set of internal fetal views by volume rendering and post acquisition slice selection. Internal views are nearly similar to those obtained during open-heart surgery. In practice, from typical long axis and apical views, two volumes are stored for further examination. Fetal movement, fetal spine and maternal obesity are unfavorable circumstances for useful examination. The routine use of cardiac volume sampling for malformations screening should be possible with increasing speed in image acquisition and in data storage to display real-time 3D imaging. PMID- 10804368 TI - [3 dimensional Doppler ultrasonography of the uterus and ovary]. PMID- 10804369 TI - [Contribution of MRI in the evaluation of fetal malformations]. AB - MRI was first used 10 years ago for the study of fetal morphology. This non invasive technique is mainly advocated for the study of fetus brain. Principle indications for MRI are Examinations are performed at 0.5 and 1.5 Tesla. Minimal study consists of T2 sequences in the 3 dimensions and T1 in one. Maternal premedication using hydroxizyne dichlorhydrate (25mg) can be proposed to avoid fetal movements. The main advantage of MRI compared with US is to detect gyral and neuronal migration anomalies, cerebral signs, of tuberous sclerosis and recent hemorrhagic lesions. PMID- 10804370 TI - [Arterial embolization of uterine myoma: results apropos of 286 cases]. AB - OBJECTIVE: Report outcome in non-surgical treatment of symptomatic uterine myomata by particulate arterial embolization. PATIENTS: and method: Two hundred eighty-six women aged 21 to 53 years with symptomatic uterine fibroids initially programmed for surgery were studied. The size and number of myomata were determined by pelvic ultrasound. After retrograde transfemoral introduction of a 4 French catheter, the left and right uterine arteries were successively catheterized. PVA particles were injected by free flow until devascularization. RESULTS: Two hundred sixty-two patients were evaluable. Complete resolution of symptoms was obtained in 245 cases. There were 17 failures. A marked reduction in the size of the myomata was observed (60% at six months). Hemorrhage disappeared in 80% of the cases immediately. Thirteen pregnancies were observed. No recurrence were observed. Complications were rare. CONCLUSION: Particulate embolization is a new minimally invasive treatment for uterine myomata which provides a alternative to hysterectomy and can replace myomectomy in young women. PMID- 10804371 TI - [The future of retroperitoneal lymph node dissection]. AB - Over the past decades, retroperitoneal lymph node dissection has been included in most gynecology oncological surgeries. Today, its usefulness is questioned. Recent studies on cervical, ovarian and endometrial cancer have attempted to redefine the role of retroperitoneal lymph node dissection. Despite theses studies, it appears obvious that dissection only has a minor impact on survival while increasing morbidity to a certain extent. The future of these procedures appears to be related to their prognostic value. The immediate goals today would be to target and remove only the necessary lymph nodes and to decrease the morbidity associated with the procedures. To reach these goals, two new approaches are presently studied: laparoscopic dissection and sentinel lymph node identification. PMID- 10804372 TI - [Radical vaginal trachelectomy for invasive cervical cancer]. AB - The use of operative laparoscopy in gynecologic oncology has paved the way to a more conservative approach in radical treatments of some cancers, particularly cervical cancer. After the vaginal approach which shortens the hospital stay and the morbidity of a radical hysterectomy, a new fertility-preserving radical treatment has been proposed : the Radical Vaginal Trachelectomy (RVT). This technique has been used since 1991 in our institution. Until July 1999, 37 patients with early-stage cervical cancer desiring to retain their fertility were treated by a laparoscopic pelvic lymphadenectomy (LPL) and RVT. The median age was 32 (22-42), and 25 were nulliparous. Twelve were FIGO stage Ia2, 22 Ib1, 2 IIa and one Ia1 (VSI+). Most of the tumors were squamous (23), grade I (21), and the lesion was 2cms or less in 35 patients. Median operating time was 275 minutes ( 200 for the last 10 cases), and blood loss 200cc. Complications were mainly due to the LPL (3 arterial injuries) rather than the RVT (one iatrogenic cystotomy and one parametrial bleeding). The mean follow-up is 42 months. Two patients recurred including one who had a small-cell neuroendocrine tumor. Ten patients had a total of 13 pregnancies with 7 live births. One newborn died of E.coli septicemia. Two patients had early miscarriages and 2 second trimester abortions (17 and 20 weeks). Two patients are still pregnant. LPL-RVT seems to be a good treatment modality for early-stage cervical cancer. It preserves fertility without lowering the chances of survival. PMID- 10804373 TI - [Laparoscopic surgery in gynecologic oncology]. AB - Panoramic retroperitoneal pelviscopy, introduced in 1987 was the first of the laparaoscopic operations used in the field of gyneceologic oncology. It was divised in order to enable the assessment of the pelvic lymph nodes prior to decision making in the management of patients with early cervical cancer. Starting from 1992, laparaoscopic surgery to all fields of gynecologic oncology and all the operations of the classical repertoire were transcribed in the new repertoire. This evolution is not without danger. Direct manipulation of an organ harboring a malignant tumor increases the chances of diffusion of malignant cells. Working with micro-instruments under CO(2) insufflation is likely to favor chances of dissemination. The true place of laparoscopic surgery is, as it has assuredly been since the beginning of its use, in the assessment of tumor surroundings and not in direct manipulation of the organ harboring the tumor. In the cases where imaging clearly shows regional and/or distal spread, it would be better to avoid laparoscopic dissection and retrieval. The most difficult problem in laparaoscopic onco-surgery is not the surgery itself, but in determining in which cases is can be used and in which it cannot. PMID- 10804374 TI - [New forms of hormonal contraception]. AB - Among new forms of hormonal contraception, three interesting exemples are described with a high level of effectiveness and low dosage regimen that allow improved safety and tolerance: a very low-dose estrogen-progestogen combination of ethinylestradiol and gestodene for 24-day cyclical administration; a progestogen-alone subcutaneous implant containing etonogestrel; and a levonorgestrel-releasing intrauterine system. These preparations appear to be particularly interesting as they provide additional possibilities for individualizing contraceptive therapy. PMID- 10804375 TI - [Polycystic ovary syndrome]. AB - The polycystic ovary syndrome (PCOS) is the most frequent endocrine disease in women of reproductive age. Hyperandrogenism, anovulation and metabolic syndrome are the cardinal features of PCOS. Hyperandrogenism results from a diffuse enzymatic hyperactivity at the theca-interstitial cell level. Anovulation is due to an impairment of the selection of a dominant follicle, while the number of smaller follicles is exaggerated. The molecular grounds of insulin resistance could be an increased Serine phosphorylation of the insulin receptor. The clinical classification of PCOS distinguishes three forms: the classic PCOS, where the three above mentioned features are present, the non classic PCOS and the asymptomatic PCOS, revealed by ultrasonography. Only the increased ovarian volume or surface (>11ml and> 5.5cm(2), respectively) must be viewed as a specific ultrasonic sign of PCOS. Cyproterone acetate remains the basic treatment of hyperandrogenism. The treatment of anovulation and infertility follows a consensual strategy. The insulin sensitizing treatment allows to decrease hyperandrogenism, to reverse the menstrual cycle irregularity and to obtain spontaneous or induced pregnancies. PMID- 10804376 TI - [Hormone replacement therapy after breast cancer]. AB - The use of hormone replacement therapy after breast cancer is controversial. The published epidemiological studies shown no association between HRT and recurrence of breast cancer. However, the interpretation of these results is very difficult because of numerous biases in this context of observational studies. Prospective randomised trials that examine the effects of HRT in women with a history of breast cancer are currently underway in United States and in Scandinavia. The markov approach is interesting for determining the global risk-benefit balance of HRT and for visualising the effects of different risk levels on breast cancer and to compare different therapeutic strategies. Hot flushes and functional symptoms represent the essential unresolved problem of these women. The results of all these will provide valuable data to define efficient and non-deleterious approaches to improve the well being of the women after breast cancer. PMID- 10804377 TI - [The primary events of puberty onset: hypothalamic, peripheral and environmental signalling]. PMID- 10804378 TI - [Maternal genital infections and premature delivery]. AB - The etiology of preterm birth remains elusive being probably multifactorial. Fifty years ago, genital infection was proposed as an important risk factor. Recent advances in the identification of several pathogens together with the potential role of bacterial vaginosis have generated renewed interest. However may studies suffered from methodological flaws and definitive answers are yet to come. In addition, studies looking at genital infections on pathological cervices already effaced or dilated, suggested that infection may play an additive interactive role to preterm birth. PMID- 10804379 TI - [Update on maternal-fetal infections by hepatitis C, HIV and cytomegalovirus]. AB - Maternal infections with hepatitis C virus, human cutomegalovirus and HIV and their management problems continue to raise concern in obstetrics. What should we recommend to decrease the incidence of hepatitis C virus congenital infection? We know that congenital human cytomegalovirus infection can lead to major neurological disease with sensorineuronal loss. The current lack of trusted prognostic elements to predict impact on the infected fetus makes any prenatal diagnosis questionable. Should we thus track the initial maternal antibody status and the newborn infection? For the last 10 years, HIV mother-to-infant transmission has decreased from 25% to less than 3%. The new problem is how to answer the more and more frequent request for pregnancy from an infected woman. The treatment must be adapted to the pregnancy in order to reduce the risks for the mother and the infant. All these questions emphasize the problems of viral infection which will require answers in the years to come. PMID- 10804380 TI - [Use of fetal pulse oximetry during labor]. AB - Fetal surveillance during labor is mainly based on fetal heart rate monitoring in France. In case of abnormal FHR, fetal scalp blood analysis may reduce the rate of cesarean section. Fetal pulse oximetry, a recent second-line technique of fetal monitoring could also reduce obstetrical interventions. Several maternity departments in France and other European countries are currently using this new tool routinely. Several observation studies have shown the potential interest of pulse oximetry. Randomized controlled trials will soon be able to conclude on the real benefits that can be expected from pulse oximetry during therm labor in normally grown fetuses. Other clinical situations such as meconium-stained amniotic fluid, preterm labor or fetal growth retardation remain to be explored. PMID- 10804381 TI - [Multicenter randomized trial of amnioinfusion]. AB - Meconium staining of the amniotic fluid in labor is a frequent problem that is associated with an increase in the risk of neonatal and maternal morbidity. Amnioinfusion is a simple technique that is designed to prevent neonatal and maternal morbidity associated with meconium. Preliminary studies indicate that amnioinfusion is a promising approach to the prevention of such complications of labor. However, further research is required. The primary objective of this multi centre randomized controlled study is to determine if amnioinfusion for thick meconium stained amniotic fluid results in a reduction in perinatal death or moderate to severe meconium aspiration syndrome. We will also assess the effects of amnioinfusion on other indicators of neonatal morbidity and on cesarean section. The study includes an evaluation of womens views on their childbirth experience and an economic evaluation of a policy of amnioinfusion The study will be achieved with the collaboration of approximately 50 obstetrical centres from across Canada, US, Europe, South America and South Africa. This multicentre trial will provide urgently needed information on the efficacy and effectiveness of amniofusion for the indication of meconium stained amniotic fluid. PMID- 10804382 TI - [Premature ovarian failure]. AB - The incidence of premature ovarian failure is highly variable according to the familial context and the clinical symptomatology leading to diagnosis. Different etiologic mechanisms have been identified. Currently, the proportion of idiopathic premature ovarian failure tends to decrease owing to the better recognition of genetic and immunological anomalies. Initially this pathology was considered irreversible. However, for the past twenty years, stigmas of a resumption of ovarian function have been noted regularly. The optimisation of treatments therefore relays on a precise knowledge of pathophysiologic mechanisms underly the symptomatology. PMID- 10804383 TI - [Fertility preservation by ovarian tissue cryopreservation]. AB - Female fertility may be compromised after radio or chemotherapy for malignant as well as non-malignant diseases. It could be saved by cryopreservation of ovarian tissue, since it is now well established that primordial follicles can survive freezing. Many births have been obtained in rodents with thawed ovarian fragments, after grafting or in vitro folliculogenesis. Both techniques are more difficult in large animal species, and need further development to enter clinical practice. However, storing ovarian samples is already indicated, especially in younger patients who will require restoration of their fertility in a far future. PMID- 10804384 TI - [Filiation in assisted reproduction with donor gametes]. AB - Donor-assisted reproduction entails a definitive separation between biological and social filiation. In ovocyte donation, the dissociation remains partial, only genetic, because the pregnancy maintains a biological bond. On the contrary, in sperm donation, the break-off is complete. Should the circumstances of his/her conception be revealed to the child? Despite a long practice of artificial insemination by donor (AID) the issue is still a matter of controversy. The majority of couples undergoing AID prefer maintaining the secret. But this attitude goes against the right of the child to be aware of his/her origin PMID- 10804385 TI - [Informed consent and principle of precaution]. AB - The so called principle of precaution can be applied to the medical activity according to two opposite conceptions : in a traditional way, the practitioner is acting following the line he/she presumes to be in the best interests of the patient, despite the psychological burden of this attitude. The information given to the patient is modulated accordingly. Such attitude is nowadays considered as excessively paternalistic. Following the recent Court decisions the information must now become absolutely complete, including the very serious, albeit exceptional, consequences of the treatments, in order to allow the patient to make in every circumstance an informed choice. Such an attitude leads to a very efficient protection of the practitioner. However, the consequences may be detrimental for the individual patients, as well as for the whole society. PMID- 10804386 TI - [Information by authorities on hospital performance]. AB - Information gathered in hospitals in Belgium by the authorities or their intermediaries is of three types: financial, medico-administrative, and quality of care assessment. From these computerized date bases, the public authorities can now measure socalled hospital "productivity". The aim announced by the authorities is to simultaneously combine increased economic effectiveness with improved quality of care. The use of this new bank of knowledge aims to enforce a sense of responsability to medical care. Regulation of the medical system by "medical management" can apply at various level depending on whether it works "a priori" or "a posteriori" and which or without financial incentives. There is however the risk of creating inaccuracies which must be taken into account when adjusting costs to each particular patient group. The use of all these tools should enable distinguishing between economic and qualitative objectives. PMID- 10804387 TI - [JMV, leading the way to new vascular medicine, and latest results of the readership survey]. PMID- 10804388 TI - [Pulse pressure: the missing link among factors for vascular disease]. PMID- 10804389 TI - [Horton's disease and aortic aneurysm: coincidence or causality? 5 cases]. AB - Five inflammatory aortopathies were disclosed 3 to 16 years after inaugural giant cell arteritis. Three patients were symptomatic: one aneurysm of the subrenal abdominal aorta discovered at work-up for an inferior arteriopathy, one thoraco abdominal aneurysm with a "fissuration" episode, one calcified thoraco-abdominal aortopathy suggesting dissection. In these three cases, there was a severe inflammatory syndrome with asthenia, fever, elevated erythrocyte sedimentation rate and a large inflammatory crown around the aortopathy. In the two asymptomatic cases, the diagnosis was made during the follow-up of Horton's disease, in one patient with active disease, the other late after the initial episode. Two aneurysms required surgical cure, with resection-prosthesis of the thoraco-abdominal aneurysm and revascularization of the digestive and renal arteries. In the 4 active cases, corticosteroid therapy cured the inflammatory process both on the basis of laboratory results and the involution of the periaortic crown and, in one case, the total regression of ureteral compression causing pyeloureteral dilatation on the left. The diagnosis of giant cell arteritis was confirmed histologically in the two operated cases. Extra-cervical localizations of aortic aneurysm of dissection in patients with giant cell arteritis is not a fortuitous coincidence but an association as demonstrated by the Mayo Clinic epidemiology. On the basis of these reported cases and data in the literature, the practical conclusions are: in case of aorta involvement, particularly with inflammation in subjects under 50, giant cell arteritis should be entertained as a possible diagnosis; in patients with giant cell arteritis, follow-up should include yearly thoracic radiograms to search for thoracic aorta involvement and Doppler and ultrasound explorations to identify any abdomino iliac lesions. This protocol is required to avoid the life-threatening complications of dissection or rupture of an aortic aneurysm. PMID- 10804390 TI - [Intermittent claudication and quality of life. Psychometric validation of the French version of the CLAU-S questionnaire]. AB - CLAU-S is a disease-specific quality of life (QoL) scale for patients with intermittent claudication due to peripheral arterial disease of the lower limbs. Initially developed in Germany, the scale has been translated into French according to classical forward/backward methodology. The objective of this study was to test the validity of the French version of CLAU-S. In its current format, the CLAU-S scale evaluates 5 dimensions: "Daily Living"; "Pain"; "Social Life"; "Disease-specific anxiety"; and "Mood", for a total of 47 items. The QoL scores and clinical findings in 157 patients with intermittent claudication were analysed. Validation of the convergence and discriminatory power of the scale were confirmed. Analysis of structural validity showed grouping of items in accordance with initial scale construction and a clear separation between the different dimensions. Analysis of the internal reliability of the questionnaire confirmed the good internal coherence of these dimensions. Reproducibility of the scores over time was confirmed by test-retest analysis. In addition to the actual validation of CLAU-S, a new, innovative mathematical technique called Structural Modelling Equations was used on this QoL instrument. It related the causal structure of the QoL components in peripheral arterial disease of the lower limbs (stage 2). An indisputable causal relationship was demonstrated between clinical parameters, particularly walking distance, and the QoL components. The classical symptom-deficit-disability causal structure of disease- specific QoL questionnaires was also studied. In particular, we found an essential relationship between the Daily Living dimension and, to a lesser extent, the Social Life activities and Disease- specific anxiety dimension, and QoL. The Pain dimension proved to be an explanatory variable of the other QoL components without being directly related to QoL. Finally, the Mood dimension was a resultant variable of QoL and not an explanatory variable of QoL. This study therefore enabled validation of the psychometric properties of the French version of CLAU-S and also demonstrated the complementary roles of walking distance and QoL in the management of patients with intermittent claudication. PMID- 10804391 TI - [Aorto-iliac aneurysms rupturing into the vena cava. A retrospective multicenter study]. AB - Eleven of the 28 centers participating in the French inter-university association for surgical research (AURC) collected retrospective data on abdominal aortic aneurysms ruptured into the vena cava. Thirty-nine cases were observed between 1979 and 1989. The signs and symptoms reported varied greatly. The pathognomonic triad associating aneurysm, abdominal pain and continuous abdominal murmur was found in one-third of the cases: 25.6% of the patients had signs of global heart failure. The diagnosis was preoperative in 74.4% of the cases. Mean delay of symptomatology before diagnosis was 7 +/- 12 hours. The analysis of operative procedures included 37 patients as 2 patients died before vascular repair could be attempted. Venous repair was done by direct suture in 76% of the cases. Peroperative blood loss was 4 600 +/- 3 500 ml. Perioperative mortality was 11 cases (28.2%). Cardiac and respiratory disorders were the cause of death in 54.4% of the fatal cases. PMID- 10804393 TI - [Advanced glycation end products are involved in microvascular permeability changes observed in microcirculation of diabetic rats in vivo]. AB - AIM: Since chronic hyperglycemia leads to Advanced Glycation End Products (AGE) formation, we were interested in its involvement in changes in microvascular permeability during diabetes. METHODS: Intravital microscopy was used to study changes in microvascular permeability during diabetes. The extravasation of a fluorescent macromolecular tracer (FITC-Albumin) was measured for one hour and, after computer-aided image analysis, was expressed as variations of normalized grey levels (arbitrary units). RESULTS: The extravasation of the tracer was increased in diabetic rats. An intravenous bolus of s-RAGE (extracellular domain of the AGE receptor) inhibits dose-dependently this increase (fig. 1). The increase of the extravasation of the macromolecular tracer can be inhibited when s-RAGE are administered 1 and 3 hours before the experiment, but not when administered 1/4 hour and 12 hours before experiment (fig. 2). Administration of antibodies against AGE receptors also inhibits the increase of permeability observed in diabetic rats (fig. 3). CONCLUSION: AGE interaction with their receptor is responsible for an increase in microvascular permeability. PMID- 10804394 TI - [Sural nerve neuropathy (external saphenous) linked to a disease of the small saphenous vein. Apropos of 5 cases]. AB - Five cases of sural nerve lesion linked to small saphenous vein pathology are reported. They were due to vein stripping (4 cases) and to thrombophlebitis (1 case). The diagnosis was especially delayed in the first 4 cases, as this pathological field is quite unknown. In all the cases, only electrodiagnosis allowed to assess the diagnosis and medical care was sufficient to control the pain of these patients. PMID- 10804392 TI - [Role of oxidative stress in permeability changes observed in the microcirculation of diabetic rats in vivo]. AB - AIM: Since an increase in the generation of free radicals has been shown in diabetes, we were interested on its involvement in changes in microvascular permeability during diabetes. METHODS: Intravital microscopy was used to study changes in microvascular permeability during diabetes. The extravasation of a fluorescent macromolecular tracer (FITC-Albumin) was measured for one hour and, after computer-aided image analysis, was expressed as variations of normalized grey levels (arbitrary units). RESULTS: The extravasation of the tracer was increased in diabetic rats. An intravenous bolus of probucol (an other anti oxidant) inhibits this increase (fig. 1). The increase of the extravasation of the macromolecular tracer can also be inhibited when both superoxide dismutase, (which dismutates superoxide to hydrogen peroxide) and catalase (which reacts with hydrogen peroxide to form water and molecular oxygen) are administered simultaneously (fig 2). When administered separately, these enzymes failed to inhibit the increase of the extravasation of the macromolecular tracer. Five-week treatment of diabetic rats by pentoxifyllin, a drug which inhibits leukocyte activation did not have any effect on the increase of albumin extravasation (fig. 3a). In addition, no difference was found in adherent leukocytes or in the leukocyte rolling flux between diabetic and normoglycemic rats (fig. 3b). CONCLUSION: Reactive oxygen species are responsible for an increase in microvascular permeability likely by leukocyte-independent mechanisms. PMID- 10804395 TI - [Inflammatory abdominal aortic aneurysm]. AB - A 58-years-old man, with no medical past history, was examined for abdominal pain and weight loss. An enlarged kidney could be palpated, and abdominal echography showed left hydronephrosis due to ureteral compression by abdominal aortic aneurysm. Laboratory data showed an inflammatory syndrome. CT scan suggested the diagnosis of inflammatory abdominal aortic aneurysm. The use of corticosteroids brought about the regression of the symptoms and the resolution of the ureteral obstruction, permitting aneurysmectomy and prosthesis replacement without ureterolysis. This report shows the interest of preoperative radiological diagnosis of the inflammatory character of abdominal aneurysm. For uncomplicated cases, preoperative treatment using corticosteroids could allow partial regression of the periaortic inflammation and easier surgical repair. PMID- 10804396 TI - [Uterine fibroid embolization: mature enough?]. PMID- 10804397 TI - [Diagnostic value of cyst fluid analysis in cystic lesions of the pancreas: current data, limitations, and perspectives]. AB - Diagnostic assessment and therapeutic indications in patients with cystic lesions of the pancreas are often difficult, especially since incidental detection of such lesions is increasing due to improvements in imaging techniques. Some lesions are benign (pseudocyst, serous cystadenoma) requiring conservative management and follow-up, whereas others are premalignant (mucinous cystadenoma, intraductal papillary mucinous tumor.) or frankly malignant (cystadenocarcinoma) and require surgical resection when possible. In about a third of cases, imaging findings are not sufficiently accurate in establishing the diagnosis, especially in cases of unilocular lesions. Fine needle aspiration can be considered as a second line method in uncertain cases. Aspiration should be performed in specialized services as the procedure requires careful consideration in some patients. Complications are usually rare and minor. The most accurate markers in this field are the carcinoembryonic antigen and the Ca 72.4. They allow to distinguish between a pseudocyst or a serous cystadenoma and a mucinous tumor (benign or malignant) with a 60% sensitivity and a specificity of nearly 100%. Cytological or histological analysis can provide valuable information but their accuracy seems to depend on tumor type. Genetic studies (Ki-ras.) may be possible in the future, including lesions with low cellular cyst fluid content. In conclusion, cyst fluid analysis after fine needle aspiration is a useful diagnostic tool but should always be interpreted in combination with clinical data and imaging findings. PMID- 10804398 TI - [Magnetic resonance imaging of soft tissue tumors of the hand and wrist]. AB - We will discuss the imaging characteristics of soft tissue tumors of the hand and wrist, especially the magnetic resonance imaging findings, since MRI has proven to be the gold standard for characterization of soft tissue tumors. Our series consists of 71 tumors of the hand and wrist, that were examined by magnetic resonance imaging. Fourty-four lesions were benign, 7 malignant, and 20 lesions were pseudotumoral masses. The signal characteristics often allow to make a correct diagnosis or to narrow down the list of differential diagnoses. It also allows to determine the extent of the lesion more accurately. PMID- 10804399 TI - [Ultrasound foetal biometry in the town of Ouagadougou, Burkina Faso. Report of a cohort of 126 pregnant women]. AB - PURPOSE: This inquiry on the ultrasound fetal biometry in the town of Ouagadougou aimed to study the fetal growth through the ultrasound biometry in order to compare it with other authors's works and to lay down a local chart. PATIENTS AND METHODS: This prospective study led from March 1997 to April 1998 The follow-up of 126 pregnant women allowed to lay down the graphs of following elements: biparietal diameter, cephalic circumference, transverse abdominal diameter, abdominal circumference and femur. RESULTS: The average age of the subjects was 26 years old+/-0.8 and 52.4% of them belonged to an average socio-economic level. All the studied parameters were closely connected with the age of pregnancy. The DAT and the abdominal circumference presented the greatest correlation (0.998). The comparison of our growth graphs with those of others authors led to notice significant differences as well as with African, European and American graphs. CONCLUSION: There is a need to lay down local charts in order to have a best estimation of the pregnancy age and the fetal growth. PMID- 10804400 TI - [Differential diagnosis of infective spondylodiscitis and erosive degenerative disk disease]. AB - PURPOSE: To assess the value of imaging in the differential diagnosis of erosive intervertebral osteochondrosis (EIVO) versus infectious discitis (ID). MATERIALS AND METHODS: Twelve cases of EIVO and 30 cases of ID were reviewed to define the usefull signs for differential diagnosis on plain films, CT, and MR. RESULTS: No single sign is sufficient, but the association of several signs is suggestive of EIVO: discal vacuum phenomenom, well-defined sclerosis and erosions of vertebral endplates, high signal strip surrounding low signal of vertebral endplates on T1 weighted images. CONCLUSION: Imaging is helpful in difficult differential diagnosis of EIVO versus ID. PMID- 10804401 TI - [Comparison between scintigraphy, B-mode, and power Doppler sonography in acute pyelonephritis in children]. AB - PURPOSE: Comparing Power Doppler imaging versus technetium-dimercapto-succinic acid (Tc-DMSA) scintigraphy in acute pyelonephritis of childhood. PATIENTS AND METHODS: INCLUSION CRITERIA: First episode of urinary tract infection, clinical and biological findings suggesting an upper lesion, absence of urological malformation or obstruction, absence of reflux (or vesico-ureteral reflux inferior to grade 3). Number of patients: 49, length of the study: 26 months (from November 95 to January 98). METHODS: Tc99m-DMSA scintigraphy (after five days), B mode and Power Doppler imaging (on the day of admission or the following day). Systematic cystography (day 5 to day 30). RESULTS: In terms of positive diagnosis, scintigraphy was superior to Power Doppler, and the latter was superior to B mode ultrasonography. Sensitivity (scintigraphy being the gold standard) was equal for both B mode and Power DopplerUS imaging, but combined Power Doppler and B mode US provided improved results. CONCLUSION: Currently, the results with Power Doppler imaging are insufficient to replace DMSA scintigraphy. However, Power Doppler is a good complement to B mode US. PMID- 10804402 TI - [Audit on pelvimetry at a polyvalent radiology department]. AB - PURPOSE: Study of the dose absorbed by the fetus during CT pelvimetry. MATERIAL AND METHODS: Twenty consecutive pelvimetries on pregnant women showed the mean fetal location in regard of the maternal pelvic skeleton (retrospective study). Then, thermoluminescent radiation detectors were put in the antropomorph phantom slices, in housing correspondent to the fetal's volume. The location of this volume has been determined by the first study. Two different CT units were tested. RESULTS: In case of cephalic presentation, the maximum fetal dose was 3 mGy. In case of pelvic presentation, because of an additional frontal topogram, the dose reached 3,3 to 3,6 mGy. CONCLUSION: We recommend: 1) reduction of parameters (120kV and 100mAs) for "normal" women; using other parameters for larger women; interval follow-up to evaluate results using these parameters; 2) tilting the bisciatic plane of section so to avoid direct exposure to the fetus; 3) limitation to 25 cm for the lateral scout; suppression of the routine frontal scout in case of pelvic presentation. PMID- 10804403 TI - [Spinal intradural hydatid cyst. A case report]. AB - The authors report the case of a 13-year-old girl in whom the diagnosis of intradural hydatid cyst was suspected on CT scan and MRI and confirmed at surgery. The interest of this case is related to the extremely rare location and its severe prognostic. CT scan and MRI can easily make an anatomic and topographic diagnosis of the intradural hydatid cyst. PMID- 10804404 TI - [Endometriosis: unusual cause of colonic obstruction]. AB - In the differential diagnosis of colonic obstruction, the endometriosis is an uncommon disease, rarely reported. The endometriosis of the bowel should be suspected on young women with a history of colonic symptoms and gynecologic complaints. The preoperative diagnosis is difficult, based on the clinical history, the imaging modalities and the endoscopic explorations. PMID- 10804405 TI - [Cervical aortic arch presenting with right recurrent palsy in an adult]. AB - We report here a very unusual case of cervical aortic arch presenting as a right recurrent paralysis in a 36-year-old woman in which MRI led to the complete diagnosis. PMID- 10804406 TI - [ [In Process Citation] PMID- 10804407 TI - [ [In Process Citation] PMID- 10804408 TI - [ [In Process Citation] PMID- 10804409 TI - [Results of lumbar and lumbosacral fusion: clinical and radiological correlations in 113 cases reviewed at 3.8 years]. AB - PURPOSE OF THE STUDY: Spinal fusion requires the use of hardware for reduction and stabilization. We present the clinical and radiological behavior of a population of patients with lumbar and lumbosacral spinal fusion. MATERIALS AND METHODS: Between 1990 and 1992, 113 patients were operated for lumbar and lumbosacral fusion. Mean age of the population was 43 years and mean follow-up was 3.8 years. Most of the fusions were L4-S1 fusions. 56% of the patients had a previous surgery. Thirteen patients in the series were reoperated and analyzed separately. In the majority of the cases, the indication for surgery was back pain with or without leg pain. Diagnoses were: spondylolisthesis, discopathy, scoliosis, and pseudoarthrosis. The spine was fused and reduced using two lordotic rods. Peroperative and postoperative lordosis were calculated on X-rays. Clinical results were analyzed with the Beaujon-Lassale score. RESULTS: Mean improvement was significantly better for spondylolisthesis than for other pathologies (85.6% versus 77.1%). Returning to work was possible for 85.5% of those with improvement and was not possible for 69.8%. The gain achieved in lordosis at surgery was lost at last follow-up. The lordosis of the construct appeared to protect against the development of discopathies above and below the construct. Discopathis led to a poor score. The rate of non-union was 7.9%, the rate of repeated surgery 6.1% and the rate of hardware removal 23.8%. At last follow-up, improvement was achieved in 45.6% of the 13 patients of the series who had repeat surgery. DISCUSSION: The results in our series are similar to those reported by others. Lumbar lordosis is an important factor: if lost, more interbody fusions may be subsequently required. Diagnosis of non-union is difficult and reoperation is the only sure manner to prove it by applying distraction-compression manoeuvres on the screws. All non-unions presented were symptomatic; incidence in the series was thus probably higher. Non-union and reoperation with a longer fusion are perhaps correlated with insufficient elasticity in the osteosynthesis. Optimal rod elasticity is a factor which remains to be defined. CONCLUSION: Clinical results of lumbar and lumbosacral fusions are not unsatisfactory, but in our series almost one patient out of three had to be reoperated. One of the reasons for so many reoperations is certainly hardware rigidity. Hardware was not removed without testing the fusion as this is the only means of sure diagnosis of non-union. Reoperation should not be considered a failure in this difficult surgery of back pain which requires long term surgical follow-up. PMID- 10804410 TI - [Arthroscopic debridement of full-thickness tears of the rotator cuff: a retrospective multicenter study of 283 cases with 3-year follow-up]. AB - PURPOSE OF THE STUDY: The purpose of this study was to assess clinical and radiological course of full-thickness rotator cuff tears treated by arthroscopic debridement and to discuss surgical indications according to patient demands and anatomical lesions. MATERIAL AND METHODS: This retrospective and multicentric study included patients with a minimal 3 years follow-up. The coronal extent of the supraspinatus tear was described as distal, intermediate or retracted. The sagittal extent to the infraspinatus and/or subscapularis and rotator interval was detailed. All patients were evaluated pre and post-operatively using Constant score and radiological A-P view for sub-acromial space measurement. In order to assess the influence of age, anatomical extent of the tear and follow-up, results were analyzed for 4 groups of patients according to their age: group I (19 cases less than 50 years), group II (88 cases from 51 to 60 years), group III (58 cases from 61 to 65 years) and group IV (118 cases over 65 years). Results were finally evaluated according the arthroscopic procedure (isolated acromioplasty and biceps tenotomy, or association of both). RESULTS: 283 patients, aged 63.1 years (range 32 to 82) entered this study. A complete antero-posterior tear of the supra spinatus tendon was noticed in 93.2 p. 100 of cases. The tear extended to the infraspinatus tendon in 57.3 p. 100, and to the subscapularis tendon in 29 p. 100 of cases. The long head of the biceps was involved in 73 p. 100 of cases. 218 acromioplasties and 116 biceps tenotomies were performed. The 2 procedures were combined in nearly 25 p. 100 of cases. At final revision (mean follow-up of 4.7 years), 98 patients (34.6 p. 100) were very satisfied, 122 (43.1 p. 100) satisfied, 43 (15.2 p. 100) moderatly satisfied and 20 (7.1 p. 100) were disappointed. The average Constant score increased from 42.4 to 67.6 points. The radiological study showed a 1 mm narrowing of the sub-acromial space. Objective results were surprisingly good for group I at a 5.4 years follow-up but dropped from group II to group IV, mainly in relation with the extent of the tear. A deleterious effect of the isolated biceps tenotomy was demonstrated in case of subacromial space superior to 7 mm. Inversely after this procedure patients with a narrowed space improved. DISCUSSION: Even if traditional bias of retrospective and mullticentric study could not be avoided, the large number of patients and long follow-up of this study are superior to most of the published papers. The extent of the supraspinatus tear seems to progress during the 50-60 years decade. Objective results appeared to be much more related to the anatomical lesions than to the arthroscopic procedure. CONCLUSION: This series did not demonstrate a secondary clinical or radiological deterioration after an endoscopic treatment without repair of full-tickness rotator cuff tears. Subacromial decompression and/or biceps tenotomy indications depend on age and extension of the tear. PMID- 10804411 TI - [Lesions of the radial nerve in fractures of the humeral diaphysis. Apropos of 62 cases]. AB - PURPOSE OF THE STUDY: Evaluate outcome of different techniques used for radial nerve repair after humeral shaft fractures. Identify situations where nerve exploration is required or inversely unneeded. Evaluate the risk of more systematic nerve exploration in case of radial nerve palsy. MATERIAL AND METHODS: Over a 10 year period, we cared for 62 patients with humeral fractures associated with radial nerve palsy who were addressed directly to our unit or referred after prior care. There were 54 men (mean age 30 years, 17 - 66) and 8 women (mean age 40 years, 19 - 80). There were 40 fractures of the mid-third and 22 fractures of the distal third of the humerus. Anatomic severity varied although most fractures were displaced or subsequent to high-energy trauma. Orthopedic treatment was used alone in 14 cases. There were 6 plate fixations and 42 intramedullary nailings. Twenty-four of the 62 cases of radial nerve palsy resulted from nerve tears but a continuous nerve was found in 38 cases. Tears were treated by primary suture (n =4), secondary graft (n =17, 5 with 2 torons and 12 with 3 or 4 torons), or primary muscle transfer (n =3). Neurolysis was used in 6 of the 38 cases with continuous nerve. Motor outcome was assessed using composite criteria taking into account the physiological role of each muscle group. RESULTS: All primary sutures gave excellent results. Excellent or good outcome was achieved in 12 of the 17 secondary grafts although a complementary transfer was required in 2 cases. Conversely, outcome was average in 2 cases and a failure in 3. Outcome depended on the length of the graft and the number of torons used. The importance of a proximal resection in healthy tissue sometimes required a complementary medial access and a graft between the biceps and the brachialis. Recovery was achieved in 37 of the 38 lesions with continuous nerve including 31 cases which did not require a second procedure. DISCUSSION: In case of nerve rupture, the outcome of primary suture appears to be better than that of secondary grafts. The success of the graft depends on the length of the tissue loss after sectioning in healthy tissue and on the number of torons used. This difference in prognosis points out the usefulness of systematic exploration in order to use primary or early direct suture in cases of high-energy displaced fractures. Minimally displaced fractures often are accompanied by a simple contusion with spontaneous recovery. However, in intermediary cases, the decision to make a systematic nerve exploration during the osteosynthesis procedure (aimed at early mobilization) is based on the fact that recovery was achieved in 23 cases in which initial exploration of a continuous nerve has been performed. Indirectly, the favorable outcome of postoperative paralyses reported by several authors, also supports this policy. PMID- 10804412 TI - [Results of diaphyseal shortening of the radius in the treatment of Kienbock's disease: a series of 31 cases]. AB - PURPOSE OF THE STUDY: Several therapeutic options have been proposed for the treatment of Kienbock's disease, including shortening of the radius. We report results in a series of 31 cases treated by diaphyseal radial shortening with a mean 6-year follow-up (range 1 - 18 years). MATERIAL AND METHODS: The series included 19 women and 12 men, mean age, 26 years (range 17 - 47 years). According to the Lichtmann classification there were 7 grade II, 23 grade IIIa and 1 grade IIIb. Mean shortening was 4.5 mm, ranging from 3 to 6 mm. Pain, mobility, muscle force and radiological presentation were assessed. RESULTS: Pain relief was achieved in all patients; 20 patients were entirely pain free and 11 had exceptional minor pain at exertion. Mean gain in mobility was 22 p. 100 both for palmar flexion and for dorsal flexion. Muscle force was improved in 30 cases with normal force in 18 and slightly less than normal force in 12. Muscle force remained insufficient in one case. According to the Michon criteria, there were 80 p. 100 excellent and good results. The radiographic assessment showed 14 improvements, 16 stabilizations, and 1 aggravation. Overall clinical ouctcome was independent of age, and preoperative radiographic aspect and length of the ulna. In addition, there was no correlation betwen radiological changes of the lunatum and the clinical result. CONCLUSION: Osteotomy for diaphyseal shortenig of the radius is a reliable simple procedure which does not require supplementary immobilization and which avoids complications encountered with self-compressive paltes. PMID- 10804413 TI - [Prevalence and morbidity of hip excentration in cerebral palsy: review of the literature]. AB - PURPOSE OF THE STUDY: Excentration of the hip is one of the main orthopedic complications of ante and perinatal cerebral palsy. Excentration can progress to dislocation even if tenotomy is performed. What is the prevalence and morbidity of hip excentration? METHODS: We reviewed the pertinent literature on the prevalence and morbidity of hip excentration in patients with cerebral palsy based on the Medline and Reedoc databases. RESULTS: Data presented in the literature vary greatly. Populations are representative of the referral network of the teams reporting the study rather than the cerebral palsy population in general. Signs reported were pain, difficult nursing procedures, difficult sitting position, pressure sores and fractures, but only one sign can be taken fully into account: 50 p. cent of the dislocated hip joints were painful. DISCUSSION: No one study gives a precise assessment of the prevalence of hip excentration and displacement and its consequences in patients with cerebral palsy. In a population of quadriplegic patients who do not walk, the question is whether complementary tenotomy should be performed, knowking the difficult operative and postoperative situation of such procedures. In this population, walking is an exceptional goal of bone surgery, pain relief and patient comfort are more common goals. Data in this literature (50 p. 100 of the dislocated hips are painful) suggest a prudent approach. However, there is no known marker which can be used to distinguish hips which will become painful from those which will remain pain free. In addition, irreversible cartilage degeneration can cause pain contraindicating reconstructive surgery. PMID- 10804414 TI - [Osteoconductive properties of bioactive glasses in a bulk form and as a coating on alumina]. AB - INTRODUCTION: Alumina on alumina friction couple has proven its reliability in the field of total hip arthroplasty. However, loosening of the alumina socket has been responsible for most of the failures. An improvement of the bone/alumina interface could be achieved with the use of an osteconductive material as a coating on alumina. The aim of this study was to evaluate the osteconductive properties of two types of bioactive glasses as a coating on alumina substrates. MATERIALS AND METHODS: Two types of coated implants (silicate glass coated alumina - AVSi, and phosphate glass coated alumina - AVP) were evaluated in a rabbit cancellous bone model. Pure alumina implants (A) were used as negative controls and bulk glasses (silicate - VSi and original 45S5, and phosphate glasses -VP) as negative controls. Sacrifices were performed at 3, 12 and 24 weeks. The interface evaluation included histomorphometry using an image analyzer. RESULTS: Silicate glasses demonstrated high osteoconductive properties. However, non mineralized osteoid tissue was the main tissue in contact with both coated implants and bulk phosphate glasses. This tissue covered over 70 p. 100 after 24 weeks of implantation, while it was never observed around pure alumina implants after 3 weeks. DISCUSSION: Amongst the hypotheses that could explain this mineralization inhibitory process, the one involving an Al(3+) glasses contamination from Al(2) O(3) is the likeliest. The high temperature coating procedure could be responsible for alumina transformation into a more soluble phase. PMID- 10804415 TI - [Pseudarthrosis and non-union of disjunctive talar fractures]. AB - PURPOSE OF THE STUDY: Neck fractures of the talus are rare. Necrosis is the complication most often observed. Mal union or non union are rarely reported: 11 cases of such complications have been treated and are presented with results of surgical treatment. MATERIAL: Seven non unions and 4 mal unions after separation fracture of the talus, presenting with pain and deformity of the hindfoot were treated. The treatment included removal of the fibrous tissue, heel reaxation, bone grafting, peritalar joints arthrodesis. RESULTS: Bone healing was observed in 5 of 7 non unions, and the correction of the hindfoot deformity in 9 of the 11 cases. DISCUSSION: Non union or mal union after separation fracture of the neck of the talus are poorly tolerated, the patients walking with the foot in internal rotation, bearing most of the weight on the lateral border of the foot. Such poor course results from: the less mechanical stiffness of the medial part of the talus, the severity and the comminution of the fracture, insufficienct reduction and fixation, too early weight bearing, and is associated with degenerative changes of peritalar joints. Functional and antomical good results can be obtained with bone healing, hindfoot reaxation and peritalar arthrodesis. We emphasize the interest of X Rays in the examination and the adequacy of a good initial treatment. PMID- 10804416 TI - [Tendon transfers in postoperative hallux varus. Apropos of 12 cases]. AB - PURPOSE OF THE STUDY: Iatrogenic hallux varus deformity after bunion surgery is a post-operative condition in which the great toe is oriented in medial deviation in relation to the first metatarsal head. This overcorrection can be a disabling complication with poor cosmetic appearance. A review of the surgical literature showed a reported hallux varus incidence range of 2 p. 100 to 13 p. 100. Various surgical interventions to treat hallux varus are described in the literature and we present here our experience in operative management of acquired hallux varus based on the results of surgical repair with the Johnson (extensor hallucis longus tendon) EHLT transfer procedure and the Hawkins (abductor hallucis tendon) procedure in 12 patients. MATERIAL AND METHODS: All of the patients suffered from painful iatrogenic hallux varus and conservative treatment was unsuccessful in all cases. The mean duration of follow-up was 42 months (range 8-96). The average age was 37 years (range: 17 to 58). The index procedure in eleven patients was a Mc Bride bunionectomy and in one case a Petersen procedure. On physical examination, a claw deformity of the hallux characterized by extension of the metatarsophalangeal joint and flexion of the interphalangeal joint was noted in all feet. Eight transfers of the abductor hallucis tendon and four complete transfers of the extensor hallucis longus tendon with interphalangeal joint fusion were done. At the time of the final follow-up, patients were interviewed and graded using a standardized questionnaire based on pain, metatarso-phalangeal range of motion, alignment of the hallux, footwear requirements, walking capabilities and first ray rest. RESULTS: Eleven patients reported satisfactory results and one patient a fair result. Overcorrection was a potential complication and in this retrospective study, one bunion deformity recurred but was asymptomatic. On radiographic evaluation the hallux abductus angle was improved by a mean 26.6 degrees and the average first intermetatarsal angle was 7 degrees (range: 3 degrees to 12 degrees ). A narrowing of the metatarso phalangeal joint was observed in four cases (2 Johnson and 2 Hawkins). DISCUSSION: The addition of a tendon transfer to the medial soft tissue release is recommanded in most cases of hallux varus in active patients without osteoarthritis of the metatarso-phalangeal joint. Our experience with the EHLT transfer or the abductor hallucis tendon transfer demonstrated excellent correction of the hallux varus deformity; however metatarso-phalangeal joint stiffness was frequently observed. As originally described by Johnson et Spiegl, the entire extensor hallucis longus tendon was used in conjunction with an interphalangeal joint arthrodesis in the presence of a fixed and painful deformity of the hallux interphalangeal joint. If no fixed claw deformity was present an abductor hallucis tendon transfer was preferred. Onset of degenerative disease involving the metatarso-phalangeal joint in four cases may jeopardize long term survival of these surgical results. PMID- 10804417 TI - [Isolated truncular paralysis of the musculocutaneous nerve of the upper limb]. AB - Isolated palsy of the musculocutaneous nerve, terminal branch of the lateral cord of the brachial plexus, is rare. It is responsible for sensory loss of the distal forearm and weakness of elbow flexion. It occurs after shoulder or clavicle surgery, trauma (fracture, dislocation, blows on the shoulder), violent exercice or extension of the forearm, prolonged positioning of the shoulder in extension abduction-external rotation and phlebotomy. Different mechanisms such as stretching, compression or direct nerve injury are encountered. We report 5 cases with isolated musculocutaneous nerve palsy, including bilateral palsy caused by violent forearm extension. In other cases, mechanisms were an extensive stretching during surgery and compression caused by prolonged supine position. Different injury locations and causes described in literature are reviewed. PMID- 10804418 TI - [Interscapholunate carpal axial dislocation: a case report]. AB - We present a case of a patient presenting a right wrist injury due to a motor vehicule accident. X-rays and CT Scan showed transcapholunate axial dislocation with the capitate impacted between the scaphoid and the lunate without perilunate dislocation. This injury was associated to a fracture of radial styloid process, trapezium, triquetrum, hamate, and a pisotriquetral and intermetacarpal dissociation. We did not find any similar case in French or English litterature. Treatment associated closed reduction and percutaneous pinning. At one year follow-up the wrist was painless with a normal range of motion. This lesion contributes to the understanding of carpal injuries mechanism. Capitate ascension within the scapholunar joint due to an important compression force is a pathogenic factor of scapholunate dissociation. PMID- 10804419 TI - [Anterior subtalar dislocation associated with a fracture of the calcaneus: a case report and review of the literature]. AB - A new case of uncommon anterior subtalar dislocation associated with a fracture of the calcaneus is reported. Mechanism of production is described with a review of literature. The criteria of radiologic diagnosis are mentioned. Closed reduction with stabilisation by Kirschner wire were undergone. Prolonged physiotherapy following 6 weeks of immobilisation allowed good result. A decreased subtalar motion without pain was found at the last check up. PMID- 10804420 TI - [Use of Ilizarov mini-fixator in the treatment of congenital brachymetatarsia]. AB - PURPOSE OF THE STUDY: The main cause of short metatarsal bones (commonly the fourth metatarsal) is premature closure of the distal epiphyseal plate. We report results of treatment of brachymetatarsia using the Ilizarov mini-fixator. MATERIAL AND METHODS: This study included 10 patients (19 feet) with a shortened fourth (18 feet) and third metatarsal bone. We performed gradual bone lengthening using the Ilizarov mini-fixator by distraction after osteotomy of the shortened metatarsal bone. RESULTS: Bone lengthening was effective (from 5 to 26 mm, mean lengthening 13 mm) almost without stepping out from the metatarsal formula. In terms of discomfort at walking, esthetic aspect of the foot, and shoe wearing, good clinical and cosmetic outcome was achieved. Pain as well as callosities on the plantar aspect of the foot disappeared in all patients. DISCUSSION: This technique of gradual distraction by callostasis gives sufficient lengthening for the treatment of brachymetatarsia. It overcomes the disadvantages of one-stage lengthening which include a small gain in length and neurovascular damage. This method could be recommended as the procedure of choice for solving the problem of brachymetatarsia. PMID- 10804421 TI - [Some demographic consequences of the HIV epidemic in the world: more focus on sub-Saharian Africa]. PMID- 10804422 TI - [Transmission of tuberculosis in Gironde: epidemiologic investigation by genomic analysis of Mycobacterium tuberculosis]. AB - BACKGROUND: Over the past few years, epidemiologic surveys of tuberculosis have been strengthened by new biologic technology, in particularly using RFLP (Restriction Fragment Length Polymorphism). This technique, which identifies Mycobacterium tuberculosis patterns, has allowed to study thoroughly tuberculosis bacilli transmission and pathogenesis. First applied on tuberculosis epidemics in at risk groups, RFLP has now an interest in the epidemiologic molecular survey of urbans populations. The aim of this study is to identify, in a French department, the proportion of clustering cases of tuberculosis, suspected of recent contamination. METHODS: An active surveillance of tuberculosis allows to record systematically the cases of tuberculosis-disease in Gironde. All M. tuberculosis isolates from the patients reported in this surveillance system were processed through IS6110 based RFLP analysis. Patients were interviewed face to face before this analysis, using a standardised data collection instrument. RESULTS: 102 patients were included in 1997; the RFLP analysis of all available strains identifies a high degree of polymorphism with 71 unique patterns; twelve groups with clustering patterns were found, grouping two (nine clusters), three (two clusters) and seven patients (one cluster) each. Those cases suspected of recent transmission were younger (age<60 years) and lived in poorer conditions. Epidemiologic links were confirmed in only 35% of the 31 patients clustered. CONCLUSION: This community survey analysis has allowed to identify at risk groups for tuberculosis transmission and to strengthen tuberculosis control in Gironde. PMID- 10804423 TI - [Incidence of the onset of disability and recovery of functional autonomy among the very old after one year]. AB - BACKGROUND: This article measures the survival rate and explores the changes in functional autonomy among both men and women over a one-year period in a sample of 272 subjects aged 80-85. METHODS: Data were taken from the Quebec Longitudinal Study on Aging, a multi-cohort prospective study focusing on health and quality of life. The sample comprises two sub-groups, namely disabled and autonomous subjects. Eligible participants were selected from the Quebec Health Insurance Plan (QHIP) file. However, to obtain the required number of participants, the sample was completed from eligible subjects who volunteered in response to TV, billboard and newspaper advertising about the project. The Functional Autonomy Measurement System (SMAF) was used to assess disability. The SMAF assesses functional disability in 5 domains: activities of daily living (ADL), mobility, instrumental activities (IADL), communication, and mental functioning. RESULTS: After one year, nearly 5% of the subjects had died and 72.5% were still autonomous, i.e., they had a score under 4.5 on the SMAF. The annual incidence of the onset of disability (including death, lengthy hospitalization, functional incapacities and cognitive deficits) was 20%. However, no difference was found between the sexes in regard to the risk of losing autonomy. The main areas where disability occurred among the previously autonomous subjects were, in order of importance, domestic tasks, mobility and communication. The study confirmed that it is possible to recover autonomy, even at this advanced age, particularly for women (11.2%), but less so for men (3.6%). CONCLUSION: This phenomenon of recovering functional autonomy suggests that octogenarians have a residual potential and confirms the effectiveness of preventive and curative measures and the benefits of interventions by community and family networks. PMID- 10804424 TI - [Perceived health and migration: a new sanitary approach?]. AB - BACKGROUND: Six percent of the French population is foreign. Paradoxically, there are in France few recent data about the health of these communities. Because a lot of different foreign communities are living in Marseille, it seemed important for us to examine the health of one of these populations. The difficulties encountered in studying the health status in the general population allow us to propose a measure based on perceived health opinion. Our first objective was to state that perceived health status of one community differs between its original country and its emigration country, taking into account the level of morbidity. Our second objective was to show that the perceived health status in the emigration population was close to the one observed in France. METHODS: Two cross sectional studies were performed in the Comoro Islands and in Marseille, a French south eastern city. The perceived health was assessed with the Duke Health Profile (DHP); the objective health was assessed by a specific questionnaire developed in the USA: the Ridit. The comparisons were conducted, taking into account the major confounding factors. RESULTS: Emigrated people relate higher perceived health status, than those living in the Comoro Islands, for the main dimensions assessed by DHP: physical (71.0 +/- 23.8 vs. 55.9 +/- 23.0), mental (74.6 +/- 19.3 vs. 50.0 +/- 18.5), general health (68.3 +/- 16.3 vs. 61.4 +/- 12.7) and perceived health (62.7 +/- 32.9 vs. 57.1 +/- 48.2). Social health score is the only one which is lower (59.2 +/- 21.8 vs. 77.6 +/- 18.3). The health status levels of the emigrated people are close to those reported in France. The variations of perceived health scores are coherent with those of the objective health indicators. CONCLUSIONS: If these results were confirmed, then this approach, based on self perceived health assessment, could be recommended to study health needs, and the management care, prevention and health promotion impact among these specific communities. PMID- 10804425 TI - A computer-assisted assessment of lifetime physical activity: reliability and validity of the QUANTAP software. AB - BACKGROUND: This study investigated the reliability and the validity of the QUANTAP (QUANTification de l'Activite Physique) interview-administered survey, a new computer-assisted tool designed to determine physical activity over a lifetime. METHODS: The tool was used to assess lifetime exercise habits in four dimensions (sport at school, leisure sport, occupation, daily activities) in 419 men and women aged 13-90 years. Physical activity indicators (time spent and energy expenditure) were calculated for 20-year periods. The inter-observer and intra-observer reliability of the tool was studied in two subgroups of 30 subjects. RESULTS: Intraclass correlation coefficients for intra-observer and inter-observer reliability varied from 0.56 to 0.96 and from 0.42 to 0.99 respectively according to the dimensions and indicators considered. Energy expenditure was not statistically significantly different from recommended nutritional intake in either males or females. Percent body fat at the time of the survey correlated with leisure sport (particularly in recent periods of practice): age-adjusted correlation coefficients varied from - 0.23 to - 0.45 among males, and from - 0.19 to - 0.31 among females. CONCLUSION: The results indicate that QUANTAP is reliable and valid to assess lifetime physical activity. It therefore provides a tool with which the long-term effects of physical activity on current health may be studied. PMID- 10804426 TI - [HIV-1 quantitative dynamics in vivo: a review of mathematical models]. AB - Over the last years, mathematical models have been applied in HIV infection to investigate the population dynamics of HIV-1 and cells of the immune system in infected hosts. They have contributed to a better understanding of the pathogenesis of AIDS. Among the model-based works, the quantitative studies carried out by two teams, during the years 1995, 1996 and 1997, have brought important results on HIV infection dynamics, reminding that HIV belonged to the lentivirus family, which had not been integrated in research of the previous years. In the studies on HIV dynamics, solutions of mathematical models were fitted to viral and/or immunological markers data in order to estimate the parameters of the viral and cellular production kinetics in infected patients. The present paper is a critical review of these studies. The methods and the most important results are presented. We explain why their impact was so considerable, but also show how the simplicity of modelling could result in conceptual errors. We finally discuss the contribution and the limits of mathematical models in the analysis of experimental data. PMID- 10804427 TI - [A review of socio-behavioural studies on adherence to antiretroviral treatments: beyond biomedical models?]. AB - This paper, based on a review of the literature of socio-behavioural research in this field, shows how the AIDS epidemic has renewed traditional approaches to patients behaviour toward medical treatment by substituting the notion of adherence for the traditional one of compliance. It shows how this issue of patients adherence has come to the forefront of HIV care with the recent diffusion of highly active antiretroviral therapeutics (HAART), because inadequate adherence has profound implications for the individual and public heath effectiveness of these therapeutic advances. The paper argues that two alternatives, and indeed conflicting, approaches to adherence to treatment in HIV infection however persist. The aim of the first approach remains to predict and correct non-adherent behaviour in certain patients and sometimes suggests that such predictions may provide justification for denying individuals treatment. This 'predictive' approach focuses on identification of individual barriers to 'good' adherence and calls on social science research to help improve the 'acceptability' of prescribed regimens for patients. An alternative 'empathic' approach focuses more on encouraging and supporting all HIV-infected patients medically eligible for HAART to devise appropriate individualised plans that can facilitate management of their treatment in their daily life. This latter approach more willingly learns from social science research which recognises the patient's subjective experience of illness as a central concern. In the future, the respective contributions of these two alternative approaches will have to be judged on the basis of their capacity to analyse both the factors which influence HIV-infected patients' initial adherence to antiretroviral treatment and those, potentially quite different, which interfere with adherence on the long term. They will also be judged for their capacity to inspire effective psychological and socio-behavioural interventions aimed at facilitating patients adherence. PMID- 10804428 TI - [Population projections: goals, methods, hypotheses]. AB - Population projections generally use the component method which consists in adapting sample size by age and sex from mortality, fertility and migration hypotheses. This exercise is not always aimed a providing projection data, but may also serve to analyze factors changing in the population. Comparatively long term demographic forecasts, particularly for mortality, can be made due to the major inertia of demographic phenomena. However, the quality of such predictions depends on the quality of knowledge of past evolution in the demographic factors and on the capacity to forecast their future evolution. PMID- 10804429 TI - [Assessment of risk factors in the case of endemic goiter in Ivory Coast]. PMID- 10804430 TI - [Resp-informations]. PMID- 10804431 TI - Effects of first myocardial infarction on left ventricular systolic and diastolic function with the use of mitral annular velocity determined by pulsed wave doppler tissue imaging. AB - This study was undertaken to assess the effect of a first myocardial infarction (MI) on the systolic and diastolic velocity profiles of the mitral annulus determined by pulsed wave Doppler tissue imaging and thereby evaluate left ventricular (LV) function after MI. Seventy-eight patients with a first MI were examined before discharge. Peak systolic, peak early diastolic, and peak late diastolic velocities were recorded at 4 different sites on the mitral annulus corresponding to the septum, anterior, lateral, and inferior sites of the left ventricle. In addition, the amplitude of mitral annular motion at the 4 above LV sites, the ejection fraction, and conventional Doppler diastolic parameters were recorded. Nineteen age-matched healthy subjects served as controls. Compared with healthy subjects, the MI patients had a significantly reduced peak systolic velocity at the mitral annulus, especially at the infarction sites. A relatively good linear correlation was found between the ejection fraction and the mean systolic velocity from the 4 LV sites (r = 0.74, P <.001). The correlation was also good when the mean peak systolic mitral annular velocity was tested against the magnitude of the mean mitral annular motion (r = 0.77, P <.001). When the patients were divided into 2 different groups with respect to an ejection fraction > or =0.50 or <0.50, a cutoff point of mean systolic mitral annular velocity of > or =7.5 cm/s had a sensitivity of 79% and a specificity of 88% in predicting a preserved global LV systolic function. Similar to systolic velocities, the early diastolic velocity was also reduced, especially at the infarction sites. The peak mitral annular early diastolic velocity correlated well with both LV ejection fraction (r =.66, P <.001) and mean systolic mitral annular motion (r = 0.68, P <.001). However, no correlation existed between the early diastolic velocity and conventional diastolic Doppler parameters. The reduced peak systolic mitral annular velocity seems to be an expression of regionally reduced systolic function. The peak early diastolic velocity is also reduced, especially at the infarction sites, and reflects regional diastolic dysfunction. Thus, quantification of myocardial velocity by Doppler tissue imaging opens up a new possibility of assessing LV function along its long axis. PMID- 10804432 TI - Myocardial perfusion and wall motion in infarction border zone: assessment by myocardial contrast echocardiography. AB - Several mechanisms have been proposed to explain the decreased wall motion (WM) at the borders of myocardial infarction (MI). We used myocardial contrast echocardiography (MCE) to investigate the relation of perfusion to WM in infarcted border zones (BZs) 6 weeks after MI in 5 sheep. After quantifying the extent of WM abnormality and the perfusion defect, normal (NL), infarcted, and BZs were defined. Peak intensity after contrast was measured in acoustic units (AU). Radiolabeled microspheres were injected to measure regional blood flow. The heart was stained with 2,3, 5-triphenyltetrazolium chloride (TTC). The perfusion defect on MCE was 33% +/- 7% of the total myocardial area and correlated well with TTC (r = 0.92, P <.03). The BZ was 8% +/- 5% of the total myocardial area. Peak intensity after contrast was decreased in MI compared with BZ and NL (MI: 2.5 +/- 1.9 AU, BZ: 8.0 +/- 3.8 AU, P <.005; NL: 10.2 +/- 6.9 AU, P <.02) and comparable in NL and BZ. The blood flow measured by microspheres was not different in NL and BZ but was decreased in MI (NL: 1.6 mL/g/min, BZ: 1.5 +/- 0.5 mL/g/min, MI: 0.7 +/- 0.5 mL/g/min; P <.0001). In this model of chronic ovine MI, the BZ was small and its perfusion was preserved. These findings support the hypothesis that tethering of normal myocardial segments explains the abnormal wall motion noted at the borders of MI. PMID- 10804433 TI - Detection of residual tissue viability within the infarct zone in patients with acute myocardial infarction: ultrasonic integrated backscatter analysis versus dobutamine stress echocardiography. AB - OBJECTIVES: The goals of this study were to analyze temporal changes in cardiac cyclic variation of integrated backscatter (CVIB) in acute myocardial infarction (AMI) and to investigate the predictive value of CVIB normalization compared with that of dobutamine stress echocardiography (DSE) in the assessment of functional recovery after revascularization. BACKGROUND: The normal CVIB is blunted by ischemia and recovers early after reperfusion, faster than wall motion improvement. Analysis of CVIB has been widely investigated for its potential to detect viable myocardium in the early stage of infarction. No studies have compared CVIB analysis with other techniques for viability assessment in patients with acute ischemic. METHODS AND RESULTS: Integrated backscatter images were obtained in 12 patients with AMI on days 1, 3, and 7 after admission and 1 month after revascularization. On day 7, DSE was performed in all patients. On admission, 22 of 144 segments were dyssynergic. On day 1, CVIB was abnormal in all 22 infarcted segments, on day 3, in 16, and on day 7, in only 10 infarcted segments. Eight of 10 segments nonviable by CVIB (CVIB-nonviable) were also nonrespondent by DSE; whereas 12 of 14 segments viable by DSE (DSE-viable) were also CVIB-viable. At follow-up, 10 CVIB-viable segments and 1 CVIB-nonviable segment showed functional recovery; whereas 10 of 14 DSE-viable segments showed functional recovery. Thus the positive predictive value of CVIB and DSE was 83% and 72%, respectively, with a diagnostic agreement between techniques in 77% of segments. CONCLUSIONS: Our data suggest that the normalization in CVIB in the first week after AMI accurately predicts residual tissue viability within the infarct zone. We also observed that the initial pattern of cyclic variation may be predictive of functional recovery. Finally, we found a good correlation between the recovery of a normal CVIB in segments that were still dysfunctional and a more validated method to assess tissue viability, such as the dobutamine test. PMID- 10804434 TI - Contrast harmonic color Doppler left ventriculography: machine-interpreted left ventricular ejection fraction compared with equilibrium-gated radionuclide ventriculography. AB - BACKGROUND: Multi-gated acquisition (equilibrium-gated radionuclide ventriculography) (MUGA) is considered the gold standard for measuring left ventricular ejection fraction (LVEF) because it is accurate, machine interpreted, and reproducible. Echocardiographic LVEF measurements are subject to variability in image acquisition and interpretation and to the limitations of 2-dimensional (2D) versus 3-dimensional imaging. GOAL: The shortcomings of traditional echocardiography may be addressed by combining multiplane 2D harmonic imaging, echocardiographic contrast, color Doppler ultrasonography, and digital image processing to create a new imaging modality: contrast harmonic color Doppler left ventriculography. METHODS: We compared the accuracy of a new method for measuring LVEF that allows for machine interpretation and uses contrast-enhanced intermittent harmonic color Doppler ultrasonography (CHCD). Quantitative LVEF measurements by hand-traced harmonic 2D echocardiography, contrast-enhanced harmonic 2D echocardiography, CHCD, and machine-interpreted CHCD were compared with MUGA in 35 patients. RESULTS: Contrast-enhanced intermittent harmonic color Doppler provided images with vivid endocardial definition in all patients, but hand-traced harmonic 2D echocardiography and contrast-enhanced harmonic 2D echocardiography had inadequate images in 9% of patients. The MUGA LVEF range was 0. 09 to 0.70. All echocardiographic methods showed excellent correlation with the MUGA LVEF (R (2) > 0.96), but the CHCD method had the best limits of agreement. CONCLUSIONS: Contrast-enhanced intermittent harmonic color Doppler LVEF correlates with MUGA at least as well as traditional noncontrasted echocardiography, but it provides diagnostic images in a greater proportion of patients. The CHCD images have vivid endocardial delineation and can be machine interpreted. PMID- 10804435 TI - The influence of heart rate on the Doppler-derived myocardial performance index. AB - The Doppler-derived myocardial performance index (MPI), defined as the sum of isovolumetric contraction and relaxation durations divided by ejection time and reflecting both systolic and diastolic myocardial function, has been found to be related to morbidity and mortality in cardiac diseases. The MPI is easily obtained, reproducible, and has a narrow range in healthy subjects. The goal of this study was to study the influence of heart rate changes on the MPI. Thirty patients with sick sinus syndrome treated with a pacemaker with a right atrial lead were studied. The patients were paced at increasing rates from 50 to 100 bpm. The MPI increased on average 0.02 +/- 0.03 per 10-bpm increase in rate. The correlation between MPI and heart rate was weak (rho = 0.31, P <.01). PMID- 10804436 TI - Usefulness of echocardiography and doppler echocardiography in endomyocardial fibrosis. AB - OBJECTIVE: Our goal was to demonstrate the usefulness of echocardiography and cardiac Doppler echocardiography (echo-Doppler) in the diagnosis of endomyocardial fibrosis, an unusual restrictive cardiomyopathy in Argentina. METHODS: Between 1980 and 1998, we studied 10 women (aged 27 to 58 years) with endomyocardial fibrosis confirmed by surgery and/or endomyocardial biopsy. Of the 10 cases of endomyocardial fibrothrombosis, 8 were biventricular and 2 were left ventricular. Six patients had only an echocardiographic study, and the last 4 patients (after 1987) had an echo-Doppler study also; 3 had a transesophageal echocardiography examination as well. Seven patients had grade III-IV dyspnea, 2 had an edematous-ascitic syndrome, and 1 had right heart failure at the first examination. Four patients died of heart failure and 1 of overimposed sepsis. Surgery was successful in 2 patients with the biventricular form of the disease. In one of them, fibrotic decortication was performed in both ventricles together with tricuspid and mitral replacement. In the other, the right side was not surgically treated because of its mild engagement. One patient was lost to follow up, and 3 patients are awaiting surgery at this writing. RESULTS: In all 10 patients, echocardiography was the first diagnostic tool used. In M-mode echocardiography, the typical image showed the "square root" sign in the septum and posterior wall in addition to the "merlon" sign, characterized by a hypercontractile basal ventricle opposing an obliterated apex. In 2-dimensional echocardiography, inversion of the normal sized heart with obliterated ventricles and dilated atria were seen in the whole group. In 1 patient, the fibrous thrombus was limited to the apex of the right ventricle (Shaper's type 1) in a biventricular form, whereas in the left side of this patient and in the other 9 patients, the fibrous thrombus that initially occupied the apex engaged the posterior papillary muscle, pulling the posterior valve downward (Shaper's type 2) and generating tricuspid and/or mitral regurgitation that was always mild or moderate. The fibrous thrombus never altered the movement of the underlying myocardium. There were hypoechoic and hyperdense echoes inside the fibrotic material (the latter compatible with calcium), and in all 10 patients, different grades of pericardial effusion were found. Echo-Doppler showed the same minimal percentage of change in mitral and tricuspid velocities as found in healthy patients, which clearly differentiates endomyocardial fibrosis from constrictive pericarditis. Furthermore, a restrictive pattern was observed on both atrioventricular valves when both sides were engaged with a markedly short tricuspid deceleration time. Pulmonary veins showed a markedly diastolic D wave and a broad reversal A wave (the latter presented a low velocity when the wall of the left atrium was diseased) caused by an increased end-diastolic left ventricular pressure to the same extent throughout the respiratory cycle. Hepatic veins showed a markedly deep diastolic forward wave throughout the respiratory cycle and a marked reversal with inspiration. CONCLUSIONS: We showed (1) echocardiographic studies of a significant number of patients with this unusual disease, (2) the characteristic diagnostic signs in M-mode and 2-dimensional echocardiography, and (3) the common echo-Doppler patterns shared by all subjects studied with this technique. PMID- 10804437 TI - Volumetric blood flow measurement with the use of dynamic 3-dimensional ultrasound color flow imaging. AB - We describe a new method for measuring blood volume flow with the use of freehand dynamic 3-dimensional echocardiography. During 10 to 20 cardiac cycles, the ultrasonographic probe was slowly tilted while its spatial position was continuously recorded with a magnetic position sensor system. The ultrasonographic data were acquired in color flow imaging mode, and the separate raw digital tissue and Doppler data were transferred to an external personal computer for postprocessing. From each time step in the reconstructed 3 dimensional data, one cross-sectional slice was extracted with the measured and recorded velocity vector components perpendicular to the slice. The volume flow rate through these slices was found by integrating the velocity vector components, and was independent of the angle between the actual flow direction and the measured velocity vector. Allowing the extracted surface to move according to the movement of anatomic structures, an estimate of the flow through the cardiac valves was achieved. The temporal resolution was preserved in the 3 dimensional reconstruction, and with a frame rate of up to 104 frames/s, the reconstruction jitter artifacts were reduced. Examples of in vivo blood volume flow measurement are given, showing the possibilities of measuring the cardiac output and analyzing blood flow velocity profiles. PMID- 10804438 TI - Aortic intramural hematoma during coronary angioplasty: insights into the pathogenesis of intramedial hemorrhage. AB - We report a case of a 74-year-old woman who had an aortic intramural hematoma as a complication of percutaneous coronary angioplasty. Transesophageal echocardiography enabled the diagnosis of aortic intramural hematoma and was very useful in the patient's management and follow-up. PMID- 10804439 TI - Echocardiographic characterization of a rare type of coronary artery fistula draining into superior vena cava. AB - We describe an incidental finding of a rare type of anomalous coronary artery originating from the right coronary sinus of Valsalva and draining into the superior vena cava. This was suspected on transthoracic echocardiography but was further clarified with the use of coronary angiography and transesophageal echocardiography. Echocardiography was a major tool for delineating the origin of the fistula, its complicated course, and the drainage site. PMID- 10804440 TI - Persistent hypoxemia occurring as a complication of tricuspid valve endocarditis. AB - A 33-year-old woman had intravenous drug-associated tricuspid valve infective endocarditis. Despite resolution of septic pulmonary emboli, hypoxemia persisted. We report a case of right-to-left shunting across a previously insignificant patent foramen ovale documented by contrast transesophageal echocardiography. Although a rare complication of tricuspid endocarditis, clinicians should be aware of this potential correctable complication. PMID- 10804441 TI - Metastatic pancreatic adenocarcinoma to the heart diagnosed antemortem. AB - Pancreatic tumors frequently metastasize widely, though it is rare to diagnose pancreatic cardiac metastases in the antemortem state. We report an unusual case of metastatic pancreatic adenocarcinoma to the right atrium. Transesophageal echocardiography showed that the tumor was attached to the superior aspect of the right atrium, prolapsing through and obstructing the tricuspid valve in diastole and retracting back into the right atrium during systole. The tumor was excised, and histologic examination confirmed the presence of moderately differentiated adenocarcinoma with a papillary architectural pattern and with desmoplastic stroma, features comparable to the original primary pancreatic neoplasm. PMID- 10804442 TI - Disk embolization of a Bjork-Shiley convexo-concave mitral valve: a cause of sudden cardiovascular collapse and mesenteric ischemia. AB - Strut fracture and disk embolization of a Bjork-Shiley convexo-concave valve is uncommon, but it should always be considered as a cause of sudden cardiovascular collapse in patients with such valves. Recognition of this clinical scenario is essential, given the importance of early diagnosis and the prevalence of these valves worldwide. We present a fatal case of disk embolization of a mitral prosthesis presenting with cardiogenic shock and mesenteric ischemia. PMID- 10804444 TI - Blood pressure monitoring begins its fifth year PMID- 10804443 TI - Echocardiographic features of genetic diseases: part 5. Tumors. PMID- 10804445 TI - Ambulatory monitoring and self-monitoring of blood pressure during pregnancy. PMID- 10804446 TI - Arterial distensibility in subjects with white-coat hypertension with and without diabetes or dyslipidaemia: comparison with normotensives and sustained hypertensives. AB - BACKGROUND: Arterial distensibility can be assessed by measuring pulse-wave velocity (PWV). OBJECTIVE: To determine whether diabetes, smoking and dyslipidaemia were associated with greater than normal stiffness of aortic walls in subjects with white-coat hypertension. METHODS: Arterial distensibility was assessed by automatic measurement of carotid-femoral PWV in 35 healthy normotensives, 46 white-coat hypertensives (WCH, clinic blood pressures >140/90 mm Hg, daytime blood pressures <130/85 mm Hg) and 81 ambulatory hypertensives (clinic blood pressures >140/90 mmHg, daytime blood pressures > or =130 mm Hg systolic or > or =85 mm Hg diastolic, or both) all matched for age, sex and body mass index. Nineteen normotensives (subgroup A), 28 WCH (subgroup A) and 37 ambulatory hypertensives (subgroup A) had only one or no other major cardiovascular risk factor whereas 16 normotensives (subgroup B), 18 WCH (subgroup B) and 44 ambulatory hypertensives (subgroup B) had also some combination of non-insulin-dependent diabetes, a smoking habit and dyslipidaemia. RESULTS: Both for the WCH and for ambulatory hypertensives diabetes and dyslipidaemia (subgroups B) were associated with higher (P<0.04) PWV (11.6+/-0.3 and 12.8+/-0.3m/s, respectively) than for subgroups A (9.3+/-0.5 and 10.9+/-0.6 m/s, respectively). In contrast, PWV for WCH in subgroup A (9.3+/-0.5m/s) did not differ (P>0.35) from those for the normotensive subgroups A (9.2+/-0.3m/s) and B (9.6+/-0.4m/s). PWV was not correlated to levels of glycaemia, glycosylated haemoglobin and cholesterolaemia. CONCLUSIONS: These results suggest that, both for ambulatory hypertensives and for WCH, diabetes and dyslipidaemia are associated with an impairment of arterial distensibility that can entail a greater than normal cardiovascular risk, which might dictate a more than usually stringent treatment of concomitant risk factors and possibly of high blood pressure. In contrast, PWV in WCH of the subgroup A did not differ from those in normotensives, reinforcing the hypothesis that WCH is associated with a benign cardiovascular outcome in the absence of other cardiovascular risk factors. PMID- 10804448 TI - Proceedings from a workshop on measurement of blood pressure of the working group on blood pressure monitoring of the European Society of Hypertension, Milan, June 1999. PMID- 10804447 TI - Effect of verapamil on home self-measurement of blood pressure and heart rate by hypertensive patients. Verapamil-Frequency Research Group. AB - BACKGROUND: Achievement of a controlled blood pressure and improvement of cardiovascular risk profile are the mainstays of therapy for hypertension. OBJECTIVE: To assess the responses of heart rate and blood pressure in hypertensive patients to 240 mg/day sustained release verapamil. METHODS: We assessed the effect of 240 mg/day sustained released verapamil on blood pressure and heart rate, measured in the office and at home, in 1395 hypertensive outpatients with mild-to-moderate hypertension, who were using an Omron HEM 705 CP automatic device for self-measurement. The period of observation was 3 months. RESULTS: Blood pressure decreased both in the medical office and at the patient's home, the measurements obtained at home being lower than those found in the office. Heart rate decreased in a significant and particular way. The decrease was greatest among those patients with histories of myocardial infarction and among relatively young patients, who exhibit a tendency towards higher than normal baseline heart rates. Overall, there was a shift of the heart-rate curve towards more controlled levels clustered around heart rates between 65 and 75 beats/min. Home self-measurement showed that the data gathered by the patients at home are reliable and that, when cut-off values of 140/90 mm Hg for blood pressure are used, the percentage of patients with controlled blood pressures is 62%, whereas the percentage obtained in the medical office by the physician is 56%. If cut-off values of 135/85 mm Hg are considered for self-measurements at home, according to the VI JNC recommendations, the percentage of patients with controlled blood pressures is 25.4%. PMID- 10804449 TI - Consequences of banning mercury and the cuff controversy. PMID- 10804450 TI - Ambulatory blood pressure for special groups. PMID- 10804451 TI - Ambulatory blood pressure in clinical trials. PMID- 10804452 TI - Proposals for simplifying the validation protocols of the British Hypertension Society and the Association for the Advancement of Medical Instrumentation. PMID- 10804453 TI - Continuous finger monitoring of blood pressure: advantages, problems and perspectives. PMID- 10804454 TI - Relationship between vitamin E requirement and polyunsaturated fatty acid intake in man: a review. AB - Vitamin E is the general term for all tocopherols and tocotrienols, of which alpha-tocopherol is the natural and biologically most active form. Although gamma tocopherol makes a significant contribution to the vitamin E CONTENT in foods, it is less effective in animal and human tissues, where alpha-tocopherol is the most effective chain-breaking lipid-soluble antioxidant. The antioxidant function of vitamin E is critical for the prevention of oxidation of tissue PUFA. Animal experiments have shown that increasing the degree of dietary fatty acid unsaturation increases the peroxidizability of the lipids and reduces the time required to develop symptoms of vitamin E deficiency. From these experiments, relative amounts of vitamin E required to protect the various fatty acids from being peroxidized, could be estimated. Since systematic studies on the vitamin E requirement in relation to PUFA consumption have not been performed in man, recommendations for vitamin E intake are based on animal experiments and human food intake data. An intake of 0.6 mg alpha-tocopherol equivalents per gram linoleic acid is generally seen as adequate for human adults. The minimum vitamin E requirement at consumption of fatty acids with a higher degree of unsaturation can be calculated by a formula, which takes into account the peroxidizability of unsaturated fatty acids and is based on the results of animal experiments. There are, however, no clear data on the vitamin E requirement of humans consuming the more unsaturated fatty acids as for instance EPA (20:5, n-3) and DHA (22:6, n-3). Studies investigating the effects of EPA and DHA supplementation have shown an increase in lipid peroxidation, although amounts of vitamin E were present that are considered adequate in relation to the calculated oxidative potential of these fatty acids. Furthermore, a calculation of the vitamin E requirement, using recent nutritional intake data, shows that a reduction in total fat intake with a concomitant increase in PUFA consumption, including EPA and DHA, will result in an increased amount of vitamin E required. In addition, the methods used in previous studies investigating vitamin E requirement and PUFA consumption (for instance erythrocyte hemolysis), and the techniques used to assess lipid peroxidation (e.g. MDA analysis), may be unsuitable to establish a quantitative relation between vitamin E intake and consumption of highly unsaturated fatty acids. Therefore, further studies are required to establish the vitamin E requirement when the intake of longer-chain, more-unsaturated fatty acids is increased. For this purpose it is necessary to use functional techniques based on the measurement of lipid peroxidation in vivo. Until these data are available, the widely used ratio of at least 0.6 mg alpha-TE/g PUFA is suggested. Higher levels may be necessary, however, for fats that are rich in fatty acids containing more than two double bonds. PMID- 10804455 TI - Retinol equivalence of carotenoids can be evaluated by hepatic vitamin A content. AB - The present study demonstrates a new method to evaluate the bioavailability of carotenoids based on the calculation of the hepatic retinol contents. Weaning male rats of Wistar strain were divided into 5 groups. Each group respectively received retinol acetate (2000-10,000 IU per kg diet), alpha-carotene (2400-6000 micrograms per kg diet), beta-carotene (2400-6000 micrograms per kg diet), mixture of alpha- and beta-carotenes in the ratio of 1:2 (2400 and 4800 micrograms per kg dit), and palm-carotene oil (2400-6000 micrograms per kg diet). The derived retinol equivalences of each carotenoid calculated according to the hepatic retinol contents were almost constant regardless of the volume of respective intake (alpha-carotene: 1.25 micrograms per IU; beta-carotene: 0.59 microgram per IU; mixture of alpha- and beta-carotene in the ratio of 1:2: 0.96 microgram per IU; Palm-carotene oil: 1.23 micrograms per IU). The results suggest that the hepatic retinol contents can be used as a new measure to evaluate the vitamin A bioavailability of carotenoids. PMID- 10804456 TI - Vitamin intake of 1- to 18-year-old German children and adolescents in the light of various recommendations. AB - Data on the vitamin intake during childhood and adolescence are rare in Europe. Here, age and sex specific percentiles of the absolute intakes and relative densities (per MJ) of retinol, carotenoids, vitamins A, E, C, B1, B2, B6, niacin and folate in a sample of 627 subjects between the age of 1 and 18 years are reported and compared to the actual recommendations from Germany, the EC and the USA. The evaluation of the intakes clearly depends on the reference value chosen. In total, the vitamin intake can be assessed to range between satisfactory and generous with the exception of folate which ranged consistently below the references. PMID- 10804457 TI - Changes in thiamin, riboflavin, niacin, beta-carotene, vitamins, C, A, D and E status of French Elderly Subjects during the first year of institutionalization. AB - Vitamin status was assessed in 26 recently institutionalized elderly subjects by combining dietary and biochemical measurements of thiamin, riboflavin, niacin, beta-carotene, vitamins C, A, D and E at admission (P1), and 1.5 (P2), 3.0 (P3), 4.5 (P4), 6.0 (P5), 12 (P6) months later. At admission, except for vitamin A, mean vitamin intakes were lower than the 1992 French Recommended Dietary Allowance. Thiamin, vitamins C, A and E status seemed nearly satisfactory as less than one-fourth of the population sample had blood values lower than the cut-off point for thiamin (erythrocyte thiamin pyrophosphate < 0.17 mumol/l), vitamin A (serum retinol < 1.05 mumol/l), vitamin C (serum vitamin C < 11.3 mumol/l) and vitamin E (serum alpha-tocopherol < 9.3 mumol/l) or higher than the cut-off point for thiamin (erythrocyte transketolase activity coefficient > 1.19). Almost half of the subjects for riboflavin, and almost all non supplemented subjects for vitamin D were in risk of vitamin deficiency (46% had an erythrocyte glutathione reductase activity coefficient > 1.19 and 72% had a plasma 25(OH)D3 < 25 nmol/l). During the study, vitamins status remained unchanged for riboflavin, niacin, vitamins A, D and E, improved for vitamin C (P = 0.004) or impaired for thiamin (P = 0.008). Thus, institutionalization seemed to have no effect on riboflavin, niacin, vitamins A, D and E status and a slight effect on thiamin and vitamin C status. PMID- 10804458 TI - Monitoring iodine following consumption of iodized salt in Tehrani inhabitants. AB - As the production, distribution and consumption of iodized salt has increased in recent years, this study was carried out to assess iodine status in Tehran in 1996. 1146 families comprising 5140 subjects in the twenty districts of Tehran city from all age groups were randomly selected. Thyroid size was examined by palpation and graded according to the WHO classification. In 163 families selected randomly, thyroid size was determined by ultrasonography and urinary iodine was measured by digestion method. Serum T4, T3 and TSH (IRMA) concentrations were also assayed by kits. Percentage of grades 1 & 2 goiter were 44 & 44% in females and 49 & 33% in males respectively. Median urinary iodine was 17.5 micrograms/dl. Mean serum T4, T3 and TSH were 8.41 +/- 1.4 micrograms/dl, 170 +/- 37 ng/dl and 1.4 +/- 0.8 mu/ml, respectively. In 118 children aged 6-10 years median urinary iodine was 17.5 micrograms/dl. Thyroid volume in children was 4.3 +/- 1.9 ml. No correlation was established between the thyroid volume and goiter grade. This study points to the adequacy of iodine intake in the majority of families residing in Tehran. PMID- 10804459 TI - Pregnancy reduces arachidonic and docosahexaenoic in plasma triacylglycerols of Korean women. AB - Plasma triacylglycerol (TG) fatty acid composition of healthy non-pregnant and non-lactating women, expectant mothers and their term neonates from Seoul, South Korea was investigated. They were ethnically homogeneous and the women had comparable age and parity. Blood samples were obtained at recruitment, third trimester and birth from the non-pregnant women, expectant mothers and neonates respectively. Percent levels of linoleic and arachidonic acids and n-6 metabolites and total n-6 were significantly lower in the pregnant women than in the non-pregnant (p < 0.0001). Similarly, there were lower levels of alpha linolenic (p = 0.033), eicosapentaenoic and docosahexaenoic acids and n-3 metabolites and total n-3 (p < 0.0001) in the expectant mothers. Compared with their mothers, the neonates had higher proportions of dihomo-gamma-linolenic, arachidonic, docosatetraenoic and docosapentaenoic acids and n-6 metabolites (p < 0.0001) and lower linoleic acid (p < 0.0001). Of the n-3 series, alpha-linolenic acid was lower (p < 0.0001) and eicosapentaenoic, docosapentaenoic and docosahexaenoic acids and n-3 metabolites and total n-3 higher (p < 0.0001) in the neonates compared with their mothers. The study reveals that pregnancy is associated with a reduction in the relative levels of plasma triacylglycerol arachidonic and docosahexaenoic acids. Moreover, it indicates that the decline is a manifestation of selective transfer of these fatty acids to the developing fetus. PMID- 10804460 TI - Somite formation and patterning. AB - As a consequence of their segmented arrangement and the diversity of their tissue derivatives, somites are key elements in the establishment of the metameric body plan in vertebrates. This article aims to largely review what is known about somite development, from the initial stages of somite formation through the process of somite regionalization along the three major body axes. The role of both cell intrinsic mechanisms and environmental cues are evaluated. The periodic and bilaterally synchronous nature of somite formation is proposed to rely on the existence of a developmental clock. Molecular mechanisms underlying these events are reported. The importance of an antero-posterior somitic polarity with respect to somite formation on one hand and body segmentation on the other hand is discussed. Finally, the mechanisms leading to the regionalization of somites along the dorso-ventral and medio-lateral axes are reviewed. This somitic compartmentalization is believed to underlie the segregation of dermis, skeleton, and dorsal and appendicular musculature. PMID- 10804461 TI - Morphogenesis of the eggshell in Drosophila. AB - The Drosophila eggshell is a specialized extracellular matrix that forms between the oocyte and overlaying somatic follicle cells during the latter stages of oogenesis. Largely proteinaceous, the eggshell is a highly organized multilayered structure with regional specializations designed to perform a variety of functions. Production of a functional eggshell features: (1) the differentiation of subsets of follicle cells in response to ovarian signals, (2) directed migrations of the follicle cells within the developing egg chamber, (3) expression of eggshell structural genes by the follicle cells in a defined temporal and spatial order, (4) postdepositional modifications of the eggshell proteins including several temporally regulated proteolytic cleavage events, and (5) regulated trafficking of several eggshell proteins in the assembling structure. By exploiting the genetic advantages of Drosophila and using evolution as a guide, the eggshell provides an excellent experimental system to study, in vivo, molecular mechanisms used to regulate protein-protein interactions throughout the assembly of a complex extracellular architecture in a developing organism. PMID- 10804462 TI - Cell biology of cytochrome P-450 in the liver. AB - Cytochromes P-450 (P-450) are members of a multigene superfamily of hemoproteins consisting the microsomal monooxygenase system with NADPH P-450 reductase (reductase) and/or reducing equivalents. Expression of many P-450 isoforms in hepatocytes is shown to be regulated at the level of transcription through interaction between cis-acting elements in the genes and DNA-binding (transacting) factors. Some isoforms of the CYP1A, 2B, 2E, and 3A subfamilies are regulated at the posttranscriptional level. For the topology of P-450 and reductase molecules in ER membrane of hepatocytes, models from stopped flow analysis and electron spin resonance are proposed. The densities of total P-450 and reductase molecules are revealed to be high enough to support the cluster model, suggesting that about ten P-450 molecules form an aggregate and surround one reductase molecule, and therefore the two enzymes form large micelles. ER proliferation after PB administration, which had been correlated with increase in P-450 level, is shown to be probably independent of the increase in P-450 level. There are considerable discrepancies among results reported on sublobular expression of various P-450 isoforms. Causes of the discrepancies are likely to be differences in experimental conditions of histochemical detection carried out and/or in species, strain, and/or sex. PMID- 10804463 TI - Alternative protein sorting pathways. AB - The term "nonclassical protein targeting" has been used to describe those pathways that have been recently discovered and differ mechanistically from the more studied "classical pathways." Because this nomenclature is rather arbitrary in terms of cellular relevance, we have chosen to group these protein sorting mechanisms under the heading "alternative protein sorting pathways" for the purpose of this review. Many of the alternative targeting pathways described are of primary importance. For example, without retrograde transport, both membrane material and targeting machinery accumulate at distal sites in the endomembrane system, preventing anterograde transport. Further, lysosome/vacuole delivery of degradative substrates by autophagic pathways is central to the role of this organelle as a primary site for intracellular degradation. Finally, targeting through the classical CPY pathway requires the ALP pathway for delivery of the vacuolar t-SNARE Vam3p. Analysis of these alternative targeting pathways provides a more complete understanding of eukaryotic cellular physiology. PMID- 10804464 TI - Structural correlates of the transepithelial water transport. AB - Transepithelial permeability is one of the fundamental problems in cell biology. Epithelial cell layers protect the organism from its environment and form a selective barrier to the exchange of molecules between the lumen of an organ and an underlying tissue. This chapter discusses some problems and analyzes the participation of intercellular junctions in the paracellular transport of water, migration of intramembrane particles in the apical membrane during its permeability changes for isotonic fluid in cells of leaky epithelia, insertion of water channels into the apical membrane and their cytoplasmic sources in cells of tight epithelia under ADH (antidiuretic hormone)-induced water flows, the osmoregulating function of giant vacuoles in the transcellular fluxes of hypotonic fluid across tight epithelia, and the role of actin filaments and microtubules in the transcellular transport of water across epithelia. PMID- 10804465 TI - Membrane trafficking and processing in Paramecium. AB - Cellular membranes are made in a cell's biosynthetic pathway and are composed of similar biochemical constituents. Nevertheless, they become differentiated as membrane components are sorted into different membrane-limited compartments. We summarize the morphological and immunological similarities and differences seen in the membranes of the various interacting compartments in the single-celled organism, Paramecium. Besides the biosynthetic pathway, membranes of the regulated secretory pathway, endocytic pathway, and phagocytic pathway are highlighted. Paramecium is a multipolarized cell in the sense that several different pools of membrane-limited compartments are targeted for exocytosis at very specific sites at the cell surface. Thus, the method used by this cell to sort and package its membrane subunits into different compartments, the processes used to transport these compartments to specific locations at the plasma membrane and to other intracellular fusion sites, the processes of membrane retrieval, and the processes of membrane docking and fusion are reviewed. Paramecium has provided an excellent model for studying the complexities of membrane trafficking in one cell using both morphological and immunocytochemical techniques. This cell also promises to be a useful model for studying aspects of the molecular biology of membrane sorting, retrieval, transport, and fusion. PMID- 10804467 TI - Winslow Homer and Cuba PMID- 10804466 TI - Role of natural benzoxazinones in the survival strategy of plants. AB - Benzoxazinoid acetal glucosides are a unique class of natural products abundant in Gramineae, including the major agricultural crops maize, wheat, and rye. These secondary metabolites are also found in several dicotyledonous species. Benzoxazinoids serve as important factors of host plant resistance against microbial diseases and insects and as allelochemicals and endogenous ligands. Interdisciplinary investigations by biologists, biochemists, and chemists are stimulated by the intention to make agricultural use of the benzoxazinones as natural pesticides. These natural products are not only constituents of a plant defense system but also part of an active allelochemical system used in the competition with other plants. This review covers biological and chemical aspects of benzoxazinone research over the last decade with special emphasis on recent advances in the elucidation of the biosynthetic pathway. PMID- 10804468 TI - Impact of a human interest video on living-donor kidney donation rates. AB - Forty-five potential living donors participated in this study to see if a human interest video featuring living donors and recipients who had been through the transplant process would increase living-donor donation rates and knowledge about living-donor organ donation. While neither donation rates nor knowledge achieved statistical significance, the data clearly demonstrated a clinically significant (clinically relevant) increase in donation rates. PMID- 10804469 TI - Watercolor: governing the irrepressible AB - Throughout many years of painting saturniid moths in watercolor I have naturally developed ways that are idiosyncratic to me. Given the possibility that others might find my methods helpful in their own work, I have always been happy to share them. I was very pleased, therefore, when the editor asked me to write this article. Many find watercolor a daunting medium. Given certain precautions watercolor, in spite of its tendency to be headstrong and to direct itself, is actually an easy medium. From long experience, I can honestly say that there is nothing I ever attempt to do in watercolor that I cannot succeed in doing. This is not to be taken to mean, of course, that I can do everything that can possibly be done in watercolor. Probably no artist in any medium could make such a claim. But my technique is adequate to my ambitions, and what artist can ask more of his tools than that? One important aim for me at this stage of my life is to help other artists experience a similar sense of control. In the course of giving many watercolor workshops, I have developed step-by-step images that illustrate in a straightforward and easily grasped sequence all my technical procedures. These images will provide the framework of this article. PMID- 10804470 TI - Introduction: the evolution of the individual insurance market. PMID- 10804471 TI - Cost and performance: a comparison of the individual and group health insurance markets. PMID- 10804472 TI - Consumers, insurers, and market behavior. PMID- 10804473 TI - Lessons from New Jersey. PMID- 10804474 TI - An evaluation of New York's reform law. PMID- 10804475 TI - An evaluation of Vermont's reform law. PMID- 10804476 TI - Riding the bull: experience with individual market reform in Washington, Kentucky, and Massachusetts. PMID- 10804477 TI - State regulation. What have we learned so far? PMID- 10804478 TI - A regulator's perspective on other states' experiences. PMID- 10804479 TI - An insurer's perspective on reform. AB - Individual health insurance markets differ from state to state, and as a result approaches to individual market reforms need to be different. In evaluating approaches, policy makers need to remember that since the decision to purchase health insurance coverage is voluntary, the potential for adverse selection exists. In addition, rather than putting the focus of individual market reforms almost exclusively on access to health insurance for a small number of persons with high-cost health conditions, more attention needs to be put on how to decrease the number of uninsured persons. This includes making the premiums paid for individual health insurance 100 percent tax deductible, similar to those of employer-based health insurance. Finally, market reforms need to treat all types of coverage issued in the individual market the same, whether they are purchased direct for the insurer or through an out-of-state association. PMID- 10804480 TI - The politics of reform. PMID- 10804481 TI - Dynamics without change: the new generation. PMID- 10804482 TI - Economic theory, economists, and the formulation of health policy. PMID- 10804483 TI - Endocervical curettage in evaluating abnormal cervical cytology. AB - OBJECTIVE: To determine the role of endocervical curettage (ECC) in the evaluation of women with abnormal cervical cytology and following treatment for cervical intraepithelial neoplasia. STUDY DESIGN: A retrospective chart review using the records of 2,126 patients who were seen at the Colposcopy Clinic, Baylor College of Medicine, between 1980 and 1995. All patients were referred because of an abnormal cervical smear and underwent repeat Pap smear, colposcopic examination in each case with biopsies as indicated, endocervical curettage and treatment, consisting of either cryotherapy, laser vaporization or a LOOP electrosurgical excision procedure. Following treatment, Pap smears were performed on a scheduled basis, and ECC was performed annually. RESULTS: The ECC was negative in 1,849 (87%) of the women. It was abnormal in 33% of women with unsatisfactory colposcopy and 10% of women with satisfactory colposcopy. ECC was abnormal in 21% of patients with a negative biopsy result and 42% of patients who did not have a biopsy performed. There was a significant increase in abnormal ECC results with increasing age. ECC had a high positive predictive value for ectocervical disease (86%) and a high negative predictive value for endocervical disease (90%). One year following treatment, < 4% of patients with a negative cervical smear had a high grade lesion detected on ECC. CONCLUSION: The use of ECC is helpful in detecting disease missed by routine colposcopy and biopsy and is most likely to detect ectocervical disease rather than true endocervical disease. Following treatment, ECC does not appear to be significantly more reliable than the Pap smear in detecting the presence of significant residual disease. PMID- 10804484 TI - Effect of exercise on blood pressure in pregnant women with a high risk of gestational hypertensive disorders. AB - OBJECTIVE: To prospectively determine whether moderate exercise during pregnancy lowers blood pressure. STUDY DESIGN: A randomized, controlled trial with one test group and one control group. All subjects have a history of mild hypertension, gestational hypertensive disorders or a family history of hypertensive disorders. Subjects were recruited before 14 weeks' gestation. After four weeks of observation, the subjects were randomly assigned to either the exercise or control group. The exercise group visited the laboratory three times a week for 10 weeks (18-28 gestation weeks) to perform 30 minutes of exercise at Rating of Perceived Exertion level 13. RESULTS: A total of 16 pregnant women (mean age, 30 years) participated. The mean metabolic equivalent during exercise sessions was 4.7 (SD = 0.8). Blood pressure measurements were compared before and after the 10 week exercise period in the two groups. Systolic blood pressures did not change significantly, but diastolic blood pressure (DBP) in the exercise group decreased by 3.5 mm Hg, while that in the control group increased by 1.1 mm Hg. Thus, the pre-post change in DBP differed by 4.6 mm Hg between groups. Exercise treatment reduced the diastolic blood pressure to a near-significant level in the exercise group (t = 2.34, df = 7, P = .052). Percent body fat did not differ between the exercise and control groups either before or after exercise treatment. ANOVA revealed that pregnancy had a significant effect (F(1, 14) = 5.7, P = .03) on increasing the percentage of fat, but exercise treatment did not (F(1, 14) = .18, P = .68). Estimated energy expenditure in overall daily physical activities during the intervention did not differ between the two groups despite the inclusion of exercise. CONCLUSION: This study detected a strong trend that 10 weeks of moderate exercise lowered the diastolic blood pressure among pregnant women at risk of hypertensive disorders. The reductions were probably due to the effect of exercise itself, not to weight or overall daily physical activity levels. PMID- 10804485 TI - The journal club. Teaching critical evaluation of clinical literature in an evidence-based environment. AB - OBJECTIVE: To develop a curriculum in epidemiology, biostatistics and experimental design that uses a journal club format for resident instruction and to assess participants' response to this format. STUDY DESIGN: A curriculum over 12 months included 24 concepts descriptive of topics considered critical in the assessment and evaluation of literature descriptive of clinical practice and patient care. There was one two-hour session per month for 12 months. Two sets of articles were distributed for each session. The first set consisted of literature on the topics of epidemiology, biostatistics and experimental design. These concepts increased in complexity throughout the academic year, starting from simple concepts, such as P values, and progressing to more-involved and complicated concepts, such as meta-analysis. The second set of articles consisted of literature on clinical topics. It was intended to illustrate and emphasize the concepts of experimental design and statistical analysis presented in the first set of articles. These clinical articles were both contemporary and historical and were from a variety of medical disciplines, including internal medicine, obstetrics and gynecology, orthopedics, infectious disease, anesthesiology, pediatrics and psychiatry. A questionnaire was distributed to the students at the conclusion of the curriculum. RESULTS: This format was well received by both residents and staff. Eighty-five percent of the residents expressed interest in continuing the format without a major change. Fifteen percent requested restricting the clinical literature to obstetrics and gynecology. CONCLUSION: This format proved an excellent method of training residents in evaluating clinical literature and providing familiarity with concepts in epidemiology, biostatistics and experimental design. PMID- 10804486 TI - Postoperative febrile morbidity in the benign gynecologic patient. Identification and management. AB - OBJECTIVE: To identify a useful temperature threshold and occurrence frequency that might represent genuine morbidity and to identify elements of the fever evaluation that were most useful in discriminating ill patients from those with transient and inconsequential temperature elevations. STUDY DESIGN: A retrospective chart review of postoperative gynecologic patients was undertaken for a one-year period. Data regarding temperatures, frequencies and workup were collected, as were descriptive data regarding the patient and case. RESULTS: Some of the previously reported associations, such as menopausal status and insurance coverage, failed to materialize as associated with temperature increases in this study. Other associations, such as blood loss and length of surgery, were confirmed. The commonly used threshold for "fever" of 38 degrees C was not as discriminatory as a threshold of 38.3 degrees C. Frequency, as well as absolute degree of elevation, did appear to relate to level of morbidity. The most useful studies in assessing temperature elevations were the white blood cell count and chest radiography. CONCLUSION: Although prospective evaluation is difficult and much work remains to be done, our data are sufficiently compelling to suggest a minimum temperature and frequency at which the otherwise-well-appearing postoperative patient should be assessed for infection. We also found the yield sufficient to make routine ordering useful only in the case of the white blood cell count and possibly the chest radiograph. Other tests should be reserved for use if directed by the physical examination or patient history. PMID- 10804487 TI - Clinical implications of intracytoplasmic sperm injection using cryopreserved testicular spermatozoa from men with azoospermia. AB - OBJECTIVE: To investigate whether sperm obtained by testicular sperm extraction (TESE) and cryopreserved well before intracytoplasmic sperm injection (ICSI) can serve as an effective sperm source. STUDY DESIGN: The role of cryopreserved testicular spermatozoa was evaluated in a retrospective analysis of consecutive ICSI cycles using fresh or cryopreserved sperm; they were followed by prospective, planned treatment using cryopreserved sperm with a modified ICSI procedure. Sixteen men (22 cycles) with obstructive or nonobstructive azoospermia were included in the retrospective analysis. Another 25 men (29 cycles) were in the planned treatment group. Following these series, the pregnancy outcomes were compared between ICSI cycles with fresh or cryopreserved testicular sperm. RESULTS: In the retrospective phase, 14 ICSI cycles were performed using fresh sperm, with 8 using cryopreserved sperm. There were no statistically significant differences between the two groups in any outcome measure. Planned treatment with cryopreserved sperm resulted in a fertilization rate of 84% and an embryo transfer rate of 89%. Thirteen couples (44%) achieved pregnancy (five ongoing, six delivered). These rates were similar to those in the retrospective phase of the study. All couples in the planned cryopreservation group had multiple aliquots (6.5 +/- 2.1) of sperm remaining after the first cycle. CONCLUSION: Cryopreserved sperm obtained by TESE can be used as an effective sperm source in ICSI cycles. Planned cryopreservation allows multiple aliquots to be stored for use in subsequent cycles and thus avoids the need for repeat biopsies. PMID- 10804488 TI - Clinical vs. sonographic estimate of birth weight in term parturients. A randomized clinical trial. AB - OBJECTIVE: To determine the relative accuracy of clinical and sonographic birth weight estimation among term parturients (> or = 37 weeks) and to assess the performance of the two techniques in identifying newborns with weights of < 2,500 g vs. > or = 2,500 g or < 2,500 g vs. at least 4,000 g. STUDY DESIGN: The sample size for this randomized clinical trial was based on the assumption that 50% of clinical predictions are within 10% of birth weight. Thus, 700 parturients were necessary to show a difference of 10% with sonographic estimates (alpha = .05, beta = .02). Inclusion criteria were singletons with a reliable gestational age of > or = 37 weeks, admitted for delivery and with no known fetal anomalies. Physicians who were unaware of previous sonographic estimates obtained the estimates. Student t and chi 2 tests were used; relative risk (RR) and 95% confidence intervals (CIs) were calculated. Receiver-operating characteristic (ROC) curves were constructed to compare the two techniques' ability to differentiate between abnormal (birth weight < 2,500 g and > 4,000 g) and normal (2,500-3,999 g). P < .05 was considered significant. RESULTS: Over 30 months, 758 term parturients were recruited; of them, 391 had clinical estimates and 367, sonographic. The two groups were similar in gestational age, prepregnancy and intrapartum body mass index, station of the presenting part, actual birth weight and frequency of newborns with weights < 2,500 g or > or = 4,000 g. Predictions based on clinical examination were significantly more likely to be within 10% of actual weight (58%) than those derived from ultrasound examination (32%; P < .0001; RR, 1.65; 95% CI, 1.43, 1.69). The areas under the ROC curves indicated that both techniques had a similar ability to differentiate normally and abnormally grown fetuses (P > .05). CONCLUSION: Among term parturients, clinical estimates had significantly higher accuracy than ones derived sonographically. PMID- 10804489 TI - Continuous low-dose antibiotics and cerclage for recurrent second-trimester pregnancy loss. AB - OBJECTIVE: To evaluate the role of continuous low-dose antibiotics in the prolongation of pregnancy in women at risk for recurrent second-trimester loss. METHODS: Only patients with a history of previous recurrent second-trimester losses associated with failed cervical cerclages were prospectively included in the study. Patients received low-dose antibiotics until delivery. Cerclage was performed at 14-24 weeks' gestation on the basis of transvaginal sonographic findings of cervical funneling. Outcome was evaluated by weeks of pregnancy gained in the current pregnancy as compared to the previous pregnancy. RESULTS: Ten patients were eligible for study after exclusions. All 10 achieved fetal viability. Pregnancy was prolonged by a mean of 13.4 +/- 4.2 weeks beyond the previous pregnancy. This was highly statistically significant (P < .001). CONCLUSION: Continuous low-dose antibiotics prolonged pregnancy in patients with recurrent second-trimester pregnancy losses and prior failed cerclage. Randomized clinical trials are needed to confirm the role of antibiotics in these high-risk pregnancies. PMID- 10804490 TI - Subcutaneous drain vs. suture in obese women undergoing cesarean delivery. A prospective, randomized trial. AB - OBJECTIVE: To determine if subcutaneous drain or closure of the subcutaneous layer decreases the incidence of wound complications in obese women undergoing cesarean delivery. STUDY DESIGN: Seventy-six obese women undergoing cesarean delivery and with at least 2 cm of subcutaneous fat were randomized to one of three groups: group 1 had suture closure of the subcutaneous tissue, group 2 had placement of a subcutaneous closed suction drain, and group 3 had neither suture closure nor drainage. RESULTS: Wound separation occurred in 12 (15.8%), seroma in 5 (6.6%) and infection in 3 (4%). There were no reports of wound hematoma. The overall incidence of any wound complication (infection, separation, seroma, hematoma) was higher in obese women who received neither subcutaneous suture nor drain as compared to obese women who received either subcutaneous suture closure or subcutaneous drain. The incidence of major wound complications (infection or separation) was also higher in obese women who received neither subcutaneous suture or drain compared to obese women who received either subcutaneous suture closure or subcutaneous drain. CONCLUSION: The use of closed suction drainage in the subcutaneous space may reduce the incidence of postoperative wound complications in obese women who have at least 2 cm of subcutaneous fat and undergo cesarean delivery. PMID- 10804491 TI - Predicting the success of a trial of labor with a simple scoring system. AB - OBJECTIVE: To determine the applicability of a simple scoring system, by Troyer and Parisi, in predicting the success of a trial of labor among parturients with prior cesarean delivery. STUDY DESIGN: Retrospectively, all patients who underwent a trial of labor over six consecutive years were reviewed. chi 2, Fisher's exact test and analysis of variance followed by the Turkey or Dunn test were used when appropriate. P < .05 was considered significant. RESULTS: There were 263 trials of labor, of which 63% (167) ended in vaginal delivery. While 21% had a score of 0, 40%, 28% and 11% had a score of 1, 2 and at least 3, respectively. The frequency of vaginal birth was significantly different between the four groups (P < .001): 98% for a score of 0, 69% for 1, 40% for 2 and 33% for 3-4. Occurrence of cesarean delivery for cephalopelvic disproportion (2%, 24%, 39%, 56%; P < .001) or for a nonreassuring fetal heart rate tracing (0%, 7%, 21%, 11%; P < .001) was significantly different between the four groups. CONCLUSION: In our population, we confirmed the inverse relationship between the Troyer-Parisi scoring system and a successful trial of labor. PMID- 10804492 TI - Effect of oral and intravenous hydration on oligohydramnios. AB - OBJECTIVE: To investigate whether oral or intravenous hydration affects oligohydramnios in cases with normal biophysical profile scores. STUDY DESIGN: One hundred twenty-four gravidas with singleton pregnancies at or near term were identified during a 16-month period retrospectively as having oligohydramnios. A total of 50 subjects with normal biophysical profiles (8 of 10) whose labor was not induced at once fell into two nonrandom, convenience sample groups: (1) 20 who were advised to drink fluids, and (2) 30 given intravenous hydration. The remaining 74 cases with oligohydramnios consisted of 60 gravidas with normal biophysical profile scores and another 14 with low scores (< or = 6 of 10), all of whom had labor induced promptly. RESULTS: Cases with normal fetal status showed comparably favorable amniotic fluid index changes. In all, 62.5% and 44.0% demonstrated improved indices after oral and intravenous hydration, respectively; mean change in amniotic fluid index was +0.7 and +0.6. The amount of intravenous hydration did not correlate with the frequency of amniotic fluid index change (46.6% increase with intravenous volumes of < or = 2,000 mL, 40.0% with volumes > or = 2,500 mL), or with magnitude of change (average +1.3 and +0.6). CONCLUSION: Oral and intravenous hydration may correct uncomplicated oligohydramnios, but neither appears to be particularly advantageous over the other. PMID- 10804493 TI - Predicting asthma severity from allergic sensitivity to cockroaches in pregnant inner city women. AB - OBJECTIVE: To measure and compare cockroach (CR)-specific immunoglobin E (IgE) in sera from pregnant women with mild, moderate and severe asthma. STUDY DESIGN: CR IgE levels were measured in stored sera collected during the Collaborative Perinatal Project. Three matched groups of 93 women were formed: group I (mild), history of asthma but no acute exacerbation; group II (moderate), acute asthma exacerbation; group III (severe), required hospitalization for a diagnosis of status asthmaticus. ANOVA was used to compare the three means. RESULTS: Mean CR IgE paralleled prenatal asthma severity. Mean values were 6.50, 13.12 and 28.99 kU/L for groups I, II and III, respectively (P = .06). High allergen sensitivity, defined as CR IgE > 60 kU/L, was identified in 8 of the 93 study samples. The prevalence of high allergen sensitivity increased as clinical asthma became more severe. Sixty-two percent (5/8) of the high allergen sensitivity occurred in group III. CONCLUSION: There appears to be a positive correlation between sensitivity to CR allergens and asthma severity during pregnancy, and these findings support further evaluation of CR allergen sensitivity as a predictor of asthma severity in pregnancy. PMID- 10804494 TI - High-risk HPV DNA detection by Hybrid Capture II. An adjunctive test for mildly abnormal cytologic smears in women > or = 50 years of age. AB - OBJECTIVE: To determine whether high-risk human papillomavirus (HPV) DNA testing could improve the detection of cervical intraepithelial neoplasia (CIN) and cancer in older women (> or = 50 years old) with mildly abnormal results on conventional cytology. STUDY DESIGN: The study was based on 119 patients aged over 50 (median, 62; range, 50-78) referred for colposcopy with Pap smears reported as atypical squamous cells of undetermined significance (ASCUS) or low grade squamous intraepithelial lesion (LSIL) from February 1997 to September 1998. The presence of high-risk HPV DNA (including HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68) was determined with the Hybrid Capture II (Digene, Inc., Silver Spring, Maryland) microplate method. Cytologic examination by Pap smear was based on the Bethesda System and, cervical biopsy was done via colposcopy. RESULTS: High-risk HPV DNA was demonstrated in 75.6% (34/45) of patients with LSIL and 52.7% (39/74) with ASCUS. Those who were HPV positive were significantly more likely to have CIN or cancer (odds ratio, 33.40; 95% confidence interval, 11.89-93.97; P < .0001). The sensitivity of HPV assay for detection of lesions more severe than CIN 2 was 100%, specificity 64.8%, positive predictive value 66.7% and negative predictive value 100%. CONCLUSION: The addition of a high-risk HPV DNA assay to cytologic examination appears to provide excellent sensitivity and negative predictive value for early detection of high grade CIN or cancer in older women with minimally abnormal Pap smears. PMID- 10804495 TI - Peripartum cardiomyopathy presenting as lower extremity arterial thromboembolism. A case report. AB - BACKGROUND: Although venous thromboembolism has been associated with peripartum cardiomyopathy, there have been no prior reports of lower extremity arterial thromboembolism complicating cardiac failure. CASE: A 38-year-old woman, gradiva 5, para 5, presented on postpartum day 9 with left pedal parasthesia. Lower extremity angiography found acute thrombotic emboli in the left popliteal artery, right tibial artery and right peroneal artery. When respiratory decompensation ensued, a transthoracic echocardiogram revealed global hypokinesis and a left ventricular ejection fraction of 30%. The patient had an uneventful recovery after treatment with digoxin, furosemide and intravenous heparin. CONCLUSION: Lower extremity arterial thromboembolism may be the initial manifestation of peripartum cardiomyopathy. PMID- 10804496 TI - Candida albicans chorioamnionitis associated with preterm labor and sudden intrauterine demise of one twin. A case report. AB - BACKGROUND: Although cervicovaginal Candida infections occur in 20-25% of pregnancies, the incidence of ascending infection in these cases is only 0.8%, and such infection rarely causes chorioamnionitis. CASE: Sudden intrauterine fetal demise (IUFD) of twin A occurred in a diabetic primigravida presenting with a twin pregnancy and preterm labor at 33 weeks of gestation. Placental pathology and autopsy of the stillborn twin revealed extensive chorioamnionitis and fetal sepsis in the presence of Candida albicans. Twin B was unaffected. CONCLUSION: In this case, C albicans chorioamnionitis seemed to be associated with sudden IUFD. PMID- 10804497 TI - Twin gestation in a woman with a uterus didelphys. A case report. AB - BACKGROUND: Twin gestation in a uterus didelphys is a rare condition, and the preferred labor course for these patients is unclear. CASE: A woman with a uterus didelphys presented at 14 weeks' gestational age. Serial ultrasound examinations demonstrated normally growing, diamniotic, dichorionic twins situated one in each uterine horn. The prenatal course was marked by intrauterine growth retardation, elevated liver functions and significant proteinuria. At 31 weeks' gestation, labor was induced; however, due to chorioamnionitis and arrest of cervical dilatation, cesarean section was performed 23 hours after rupture of membranes, and two viable female infants (1,480 and 1,421 g) were delivered. CONCLUSION: Twin gestation in a uterus didelphys is a rare condition, in which a trial of labor may be offered, with careful monitoring of both fetuses. PMID- 10804498 TI - Reversal of Ballantyne syndrome by selective second-trimester fetal termination. A case report. AB - BACKGROUND: Ballantyne syndrome (mirror syndrome, triple edema) describes the unusual association of fetal and placental hydrops with maternal preeclampsia. In most cases, the poor fetal prognosis and associated maternal risks warrant delivery regardless of gestational age. We used novel therapy for Ballantyne syndrome in a twin pregnancy. CASE: Ballantyne syndrome occurred at 16 weeks' gestation due to severe, unexplained hydrops in one of dichorionic twins. Selective termination of the affected fetal twin resulted in reversal of the preeclamptic findings in the mother, and the surviving twin was born uneventfully at term. CONCLUSION: This case strengthens the association of fetal and placental hydrops with preeclampsia and suggests selective fetal termination for Ballantyne syndrome due to hydrops in one of multichorionic fetuses. PMID- 10804499 TI - Cesarean delivery: bladder flap and use of diathermy. PMID- 10804500 TI - Expression and purification of recombinant histones and nucleosome reconstitution. PMID- 10804501 TI - Preparation and analysis of positioned nucleosomes. PMID- 10804502 TI - Site-directed chemical probing of histone-DNA interactions. PMID- 10804503 TI - Base-pair resolution mapping of nucleosomes in vitro. PMID- 10804504 TI - Equilibrium and dynamic nucleosome stability. PMID- 10804505 TI - Nucleosome structure and dynamics. The DNA minicircle approach. PMID- 10804506 TI - Analysis of linker histone binding to mono- and dinucleosomes. PMID- 10804507 TI - Quantitative analysis of chromatin higher-order organization using agarose gel electrophoresis. PMID- 10804508 TI - Analytical ultracentrifugation of chromatin. PMID- 10804509 TI - Analysis of chromatin by scanning force microscopy. PMID- 10804510 TI - In vivo mapping of nucleosomes using psoralen-DNA crosslinking and primer extension. PMID- 10804511 TI - Preparation of chromatin assembly extracts from Xenopus oocytes. PMID- 10804512 TI - Preparation of chromatin assembly extracts from preblastoderm Drosophila embryos. PMID- 10804513 TI - A solid-phase approach for the analysis of reconstituted chromatin. PMID- 10804514 TI - Reconstitution and analysis of hyperacetylated chromatin. PMID- 10804515 TI - Assembly of mitotic chromosomes in Xenopus egg extract. PMID- 10804516 TI - Nucleotide excision repair coupled to chromatin assembly. PMID- 10804517 TI - Photolyase. A molecular tool to characterize chromatin structure in yeast. PMID- 10804518 TI - Transcriptional and structural analyses of isolated SV40 chromatin. PMID- 10804519 TI - In vitro replication of chromatin templates. PMID- 10804520 TI - Analysis of HMG-14/-17-containing chromatin. PMID- 10804521 TI - Identification and analysis of native nucleosomal histone acetyltransferase complexes. PMID- 10804522 TI - Analysis of nucleosome disruption by ATP-driven chromatin remodeling complexes. PMID- 10804523 TI - Nucleosome remodeling factor NURF and in vitro transcription of chromatin. PMID- 10804524 TI - An SDS-PAGE-based enzyme activity assay for the detection and identification of histone acetyltransferases. PMID- 10804525 TI - Analysis of DNaseI hypersensitive sites in chromatin by cleavage in permeabilized cells. PMID- 10804526 TI - Mapping of nucleosome positions in yeast. PMID- 10804527 TI - Analysis of DNA topology in yeast chromatin. PMID- 10804528 TI - DNA methyltransferases as probes for chromatin structure in yeast. PMID- 10804529 TI - Restriction nucleases as probes for chromatin structure. PMID- 10804530 TI - Genomic footprinting using nucleases. PMID- 10804531 TI - In situ analysis of chromatin proteins during development and cell differentiation using flow cytometry. PMID- 10804532 TI - Mapping DNA target sites of chromatin proteins in vivo by formaldehyde crosslinking. PMID- 10804533 TI - Mapping DNA interaction sites of chromosomal proteins. Crosslinking studies in yeast. PMID- 10804534 TI - UV laser footprinting and protein-DNA crosslinking. Application to chromatin. PMID- 10804535 TI - An in vivo UV crosslinking assay that detects DNA binding by sequence-specific transcription factors. PMID- 10804536 TI - Cord blood banking and stem cell transplantation: implications for the practicing Ob/Gyn. PMID- 10804537 TI - Fetal intracardiac echogenic foci: current understanding and clinical significance. AB - The detection of fetal intracardiac echogenic foci (ICEF) by ultrasound was first reported in 1987. Despite many investigations, the relationship of ICEF with congenital heart malformations and chromosomal abnormalities remains unclear. This review describes the current understanding of ICEF based on a literature search from 1980 to the present. ICEF are observed in 0.5 to 20 percent of fetuses, with an overall frequency of 5.6 percent. These small, discrete structures near the papillary muscles and chordae tendinae move in synchrony with the intraventricular valves. They likely represent microcalcification of the papillary muscles. ICEF are most commonly seen in the left ventricle and occasionally in the right ventricle or bilaterally. Intra-atrial or diffuse ICEF are rare. In the chromosomally normal fetus, ICEF are not associated with congenital heart defects. The presence of ICEF in fetuses at high risk for chromosomal abnormalities suggests an increased possibility of aneuploidy, especially if other sonographic markers are noted. A similar association is observed with trisomy 21 in particular. The significance of ICEF in fetuses at low risk for aneuploidy is less clear and represents an area for future research. PMID- 10804538 TI - Cervimetry: a review of methods for measuring cervical dilatation during labor. AB - Several different methods of measuring cervical dilatation have been described. In this article, we review those methods and examine findings from studies using them. Although many instruments have been developed to measure cervical dilatation during labor and their use as a research tool has been established, no device has yet been successfully used for clinical obstetrics. The ideal device has not yet been developed; however, because repeated digital cervical examinations are time consuming for the clinician, are poorly reproducible, and are uncomfortable for the patient, continued efforts to develop a cervimeter suitable for clinical use is a worthwhile endeavor. PMID- 10804539 TI - Hyperinsulinism and its interaction with hyperandrogenism in polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women of reproductive age. It has become increasingly evident that insulin resistance plays a significant role both as a cause and result of the syndrome. The purpose of this review is to summarize the possible mechanisms leading to insulin resistance and resultant hyperinsulinism (HI) and their interaction with hyperandrogenism (HA) in PCOS. We conducted a computerized search of MEDLINE for relevant studies in the English literature published between January 1966 and January 2000. We reviewed all studies that investigated the roles of insulin, insulin receptor, and insulin gene in insulin resistance and its interaction with hyperandrogenism in PCOS. Insulin resistance in PCOS seems to involve a postbinding defect in the insulin receptor and/or in the receptor signal transduction. Current research has focused on identifying a genetic predisposition for insulin resistance in this syndrome. The answer to the question whether HI or HA is the initiating event is still unclear inasmuch as there are clinical and molecular evidences to support both of these approaches. Our view is that whichever is the triggering insult, a vicious cycle is established where HI acts to aggravate HA and vice versa. In this model, obesity and genetic predisposition seem to be the independent factors that can give rise or contribute to HI, HA, or both simultaneously. It seems that "hyperinsulinemic hyperandrogenism" represents a significant subgroup of PCOS, which probably needs to be renamed and reclassified in the light of this new approach. PMID- 10804540 TI - Bacterial vaginosis in pregnancy. AB - Bacterial vaginosis is a clinical condition caused by replacement of the normal hydrogen peroxide producing Lactobacillus sp. in the vagina with high concentrations of characteristic sets of aerobic and anaerobic bacteria. Bacterial vaginosis is the most prevalent cause of vaginal discharge or malodor, although 50 percent of women who meet the criteria for this condition are asymptomatic. Bacterial vaginosis is reported in 10 to 41 percent of women, and new evidence has shown association with maternal and fetal morbidity. Studies have shown that spontaneous abortion, preterm labor, premature birth, preterm premature rupture of the membranes, amniotic fluid infection, postpartum endometritis, and postcesarean wound infections are increased because of infection with bacterial vaginosis during pregnancy. Clinical trials demonstrated important reductions in many of these adverse events with appropriate screening and antimicrobial treatment protocols. New low-cost, diagnostic, point-of-care screening tools are available for rapid screening of patients, affording the physician the opportunity to potentially make a dramatic clinical and cost impact in preventing preterm birth and the costly sequelae of prematurity. Practicing physicians need to be aware of current guidelines for screening and treating pregnant patients for bacterial vaginosis. The authors recommend that all pregnant women be screened and treated with the Centers for Disease Control and Prevention (CDC-P) recommended oral regimens early in pregnancy. Each treated women should be evaluated for "test of cure" 1 month after treatment. Mothers likely to benefit from "screen and treat" approaches include 1) those with the highest concentrations of genital anaerobes and mycoplasmas, 2) women with prior preterm birth or who have low body mass (BMI < 19.8 kg/m2), 3) those with evidence of endometritis before pregnancy, and 4) those who are treated with oral agents effective for both presumed intrauterine mycoplasmas and other bacterial vaginosis flora (i.e., oral clindamycin or erythromycin and metronidazole). PMID- 10804541 TI - Synthesis and characterization of water-soluble amino fullerene derivatives AB - [formula: see text] A series of amino-substituted methanofullerene derivatives were prepared by mono-, tris-, and hexa-Bingel-Hirsch reactions using an N protected malonate derivative. Upon scission of the protecting groups, the e,e,e tris- and octahedral Th hexa-methano amino fullerenes were found to be among the most water-soluble fullerene derivatives yet prepared. 3He NMR data on corresponding adducts of 3He/C60 helped verify the assigned structures. UV spectral studies confirmed the influence of aggregation on solubility of these adducts in water. PMID- 10804542 TI - A conformationally reliable spacer for molecular tweezers AB - [figure: see text] A reliable and rigid spacer, trans,trans,trans perhydronaphthacene, for molecular tweezers was designed. A synthesis and molecular recognition study of its 1,14-disubstituted derivatives was carried out. PMID- 10804543 TI - Opposite pi-face selectivity for the DMD and m-CPBA epoxidations of chiral 2,2 dimethyloxazolidine derivatives of tiglic amides: control by steric interactions versus hydrogen bonding AB - [formula: see text] A high extent but opposite sense in the diastereoselectivity has been observed for the DMD and m-CPBA epoxidations of the optically active tiglic amides (S)-1 with 2,2-dimethyloxazolidines as chiral auxiliaries. This unprecedented reversed pi-facial differentiation for these two peroxidic oxidants is rationalized in terms of like (lk) and unlike (ul) transition structures: For DMD, steric interactions dominate, such that the unlike transition structure is favored, while for m-CPBA, hydrogen-bonding effects overcome these steric repulsions and the like one is preferred. PMID- 10804544 TI - Intramolecular Diels-Alder cyclizations of (E)-1-Nitro-1,7,9-decatrienes: synthesis of the AB ring system of norzoanthamine. AB - [formula: see text] Cyclizations of substituted (E)-1-nitro-1,7,9-decatrienes under thermal and Lewis acid conditions have led to the formation of decalin ring systems with excellent endo selectivity. This strategy has been applied to the synthesis of the AB ring system of norzoanthamine. PMID- 10804545 TI - An efficient Pd-catalyzed route to silyl esters AB - [formula: see text] A one-step, highly selective catalytic route to silyl esters is reported. Commercially available silanes with Si-H functionality were converted to silyl or siloxy esters in the presence of Pd(OAc)2 under mild reaction conditions. This protocol was found to be equally applicable for the modification of multiple silicon centers in one framework and lead to the corresponding polysilyl esters in high yields. A comparison of catalytic efficiency of Pd(OAc)2 versus Pd on carbon was also undertaken. PMID- 10804546 TI - A tyrosine-derived benzofuranone related to diazonamide A. AB - [formula: see text] Cyclization of 8 with PPA followed by MCPBA oxidation affords the benzofuranone 10. Treatment with a chiral chloroformate in the presence of DMAP affords the target benzofuranone 13 via an enol ester 12. PMID- 10804547 TI - Progress toward synthesis of diazonamide A. Preparation of a 3-(oxazol-5-yl)-4 trifluoromethyl-sulfonyloxyindole and its use in biaryl coupling reactions. AB - [formula: see text] The synthesis of a 3-oxazol-5-yl-indole-4-triflate is described, featuring a Scholkopf reaction to prepare the oxazole. In addition, the participation of this intermediate in biaryl coupling reactions toward the total synthesis of the natural product diazonamide A is presented. PMID- 10804548 TI - Dynamic kinetic resolution of alpha-hydroxy acid esters AB - [formula: see text] Enzymatic resolution in combination with ruthenium-catalyzed racemization of the substrate led to dynamic kinetic resolution of alpha-hydroxy esters in good yields and excellent ee's. Studies of different parameters showed that the best results were obtained using Pseudomonas cepacia lipase, ruthenium catalyst 3, and 4-chlorophenyl acetate as acyl donor in cyclohexane. PMID- 10804549 TI - delta-Amino beta-keto esters, a designed polyfunctionalized chiral building block for alkaloid synthesis. Asymmetric synthesis of (R)-(+)-2-phenylpiperidine and ( )-SS20846A. AB - [formula: see text] delta-Amino beta-keto esters 3 and 11 are designed polyfunctionalized chiral building blocks for alkaloid synthesis and are prepared in one step from the corresponding sulfinimine (N-sulfinyl imine). Concise highly enantioselective four-step syntheses of 2-phenylpiperidine (7) and SS20846A (14) from 3 and 11, respectively, are described. PMID- 10804551 TI - Selective hydrogenolysis of novel benzyl carbamate protecting groups AB - [formula: see text] Highly efficient and selective hydrogenolysis of the 2 naphthylmethyl carbamate group (CNAP) in the presence of the 4 trifluoromethylbenzyl carbamate group (CTFB) has been observed for a wide range of substrates. PMID- 10804550 TI - A new route to benzo[4,5]cyclohepta[1,2-b]naphthalenes: synthesis of radermachol. AB - [formula: see text] Condensation of 3-phenylsulfonyl-1,3-isobenzofuranone (3) with benzocyclohept-6,7-en-5-ones such as 4 and 15 provides a straightforward, general method for synthesis of functionalized benzo[4,5]cyclohepta[1,2 b]naphthalenes (e.g., 5 and 16). This finding was used to achieve a brief and efficient preparation of 6,7-benzo-3,4-(1,4-dimethoxy-2,3-naphtho)-1,5 dioxosuberane (2), an established intermediate to the naturally occurring red pigment radermachol (1). PMID- 10804552 TI - Oxazoline N-oxide mediated [2 + 3] cycloadditions. Application to a formal synthesis of (+)-carpetimycin A. AB - [formula: see text] Cycloaddition between gamma,delta-unsaturated beta-enamino ester 9 and camphor-derived oxazoline N-oxide 8 afforded a single adduct, 14. Dipolarophile 9 proved to be very reactive despite the substitution on the double bond. Stereoselective sodium cyanoborohydride reduction of the imminium intermediate 14a gave rise stereoselectively to beta-amino ester derivative 15a. Oxidative acidic hydrolysis, oxidation of the resulting aldehyde 18, deprotection, and cyclization afforded the beta-lactam 23, a direct precursor of (+)-carpetimycin A. PMID- 10804553 TI - Supramolecular phthalocyanine dimers based on the secondary dialkylammonium cation/dibenzo-24-crown-8 recognition motif AB - [formula: see text] New unsymmetrically substituted DB24C8-phthalocyanines, which are able to form complexes with suitable dialkylammonium cations, have been prepared. These complexes most probably have a pseudorotaxane geometry. PMID- 10804554 TI - Desymmetrization of 1,4-dien-3-ols and related compounds via Ueno-Stork radical cyclizations AB - [formula: see text] Desymmetrization of 1,4-dien-3-ols and related compounds via Ueno-Stork radical cyclizations is reported. The stereochemistry of the cyclization is controlled by the acetal center. Excellent stereocontrol at C(4) and C(5) of the newly formed tetrahydrofuran rings is observed. Use of a chiral auxiliary allows the preparation of enantiomerically pure material. The utility of this method has been demonstrated by achieving a short synthesis of (+) eldanolide, the pheromone of the male African sugarcane stem borer Eldana saccharina. PMID- 10804555 TI - Solid-phase synthesis of carbocyclic nucleosides. AB - [formula: see text] An efficient solid-phase synthesis of carbocyclic nucleosides has been developed. The key step is the palladium-catalyzed coupling of a purine derivative to a resin-bound allylic benzoate. The resulting products may be further functionalized on the solid phase. Acidic cleavage affords carbocyclic nucleosides, a class of compounds with demonstrated biological activity and substantial current interest. PMID- 10804556 TI - Synthesis of heterocyclic thiosulfonates AB - [formula: see text] A simple synthesis of heterocyclic thiosulfonates containing indole, indoline, benzoimidazole, and quinoxaline rings is described. The synthesis of these thiosulfonates involves the preparation of the appropriately substituted thiols followed by sulfonylation to give thiosulfonates. The corresponding thiols were prepared in a simple and efficient manner by using a thiocyanation reaction either prior to heterocycle ring formation or after heterocycle ring formation. These thiosulfonates were coupled successfully to the 5,6-dihydropyran-2-one ring to give products that showed excellent HIV protease activity. PMID- 10804557 TI - Diels-Alder reaction of sugar-derived cyclic dienophiles with cyclopentadiene. Factors affecting the reactivity and stereoselectivity AB - [formula: see text] A sugar-derived cyclic dienophile showed a high exo and face selectivity in the Diels-Alder reaction with cyclopentadiene. Structural modifications on the dienophile structure were made, and the outcomes of the cycloaddition reactions were investigated. These results and the spectroscopic data recorded within a series of analogues were used to analyze the factors affecting the reactivity and exo selectivity. PMID- 10804558 TI - A trans-stereoselective synthesis of 3-Halo-4-alkyl(aryl)-NH-azetidin-2-ones AB - [formula: see text] Conrotatory ring closure of 1-halo-3-aza-4-alkyl-1,3-dienes in refluxing toluene gives rise to 3-halo-4-aryl-2-azetidinones in satisfactory yields. Dehalogenation of the resulting beta-lactams by tris(trimethylsilyl)silane furnished 3-unsubstituted azetidinones, valuable intermediates in the synthesis of biologically active compounds. PMID- 10804559 TI - Synthesis of an anti-methicillin-resistant Staphylococcus aureus (MRSA) carbapenem via stannatrane-mediated Stille coupling. AB - [formula: see text] A short synthesis of carbapenem 1 is described. They key step involves the cross-coupling of an enol triflate with an amino-substituted sp3 carbon. This cross-couping, which allows the introduction of the complete side chain in one step, utilizes a stannatrane as the heteroalkyl transfer reagent. PMID- 10804560 TI - Synthesis and UV photolysis of oligodeoxynucleotides that contain 5 (phenylthiomethyl)-2'-deoxyuridine: a specific photolabile precursor of 5-(2' deoxyuridilyl)methyl radical. AB - [formula: see text] The title exocyclic radical (2) is generated via photochemical cleavage of 5-(phenylthiomethyl)-2'-deoxyuridine (8). The latter thionucleoside (8) was successfully incorporated into DNA oligomers by automated DNA synthesis using phosphoramidite chemistry. UV exposure of 8 containing oligonucleotides under (an)aerobic conditions gives rise to specific base lesions. The photoproducts have been isolated and further characterized on the basis of detailed NMR and mass spectrometric analyses. PMID- 10804561 TI - Synthesis of unnatural amino acids via Suzuki cross-coupling of enantiopure vinyloxazolidine derivatives. AB - [formula: see text] (R and S)-alpha-Amino alcohols and alpha-amino acids, including 4-methoxyhomophenylalanine, with a variety of unnatural side chains have been synthesized via palladium-catalyzed cross-coupling Suzuki reactions. The key building blocks 1 and 2, synthesized from the common achiral precursor 2 butene-1,4-diol, were made enantiopure utilizing a Pseudomonas cepacia lipase catalyzed kinetic resolution. The optimal conditions for the Suzuki cross coupling and the subsequent oxidations of the resultant alpha-amino alcohols are described. PMID- 10804562 TI - Synthesis of carbamate-containing cyclodextrin analogues. AB - [formula: see text] The one-pot cyclooligomerization of a saccharide-derived p nitrophenyl carbamate monomer was developed to generate a series of novel carbamate-containing cyclodextrin analogues. The "transcarbamoylation" occurs by initial base-induced activation to the isocyanate, followed by polycondensation/cyclization of the isocyanato alcohol. In the presence of NaH, only cyclized oligomers were observed, suggesting the importance of Na+ in promoting the efficiency of the cyclization process. The facile deprotection of the oligomers was achieved. PMID- 10804563 TI - Highly enantioselective Michael reactions catalyzed by a chiral quaternary ammonium salt. Illustration by asymmetric syntheses of (S)-ornithine and chiral 2 cyclohexenones. AB - [formula: see text] The use of the chiral quaternary ammonium salts 1a and 1b makes possible enantioselective Michael reactions which have been applied to the asymmetric syntheses of (S)-ornithine (2) and the chiral 2-cyclohexenone 6. PMID- 10804564 TI - Palladium-catalyzed intermolecular coupling of aryl halides and amides. AB - [formula: see text] The first general intermolecular C-N bond-forming reactions between aryl halides and amides were realized using a palladium catalyst with Xantphos as the ligand. Aryl triflates, carbamates, and sulfonamides are also viable substrates for the amidations, which proceed at 45-110 degrees C with 1-4 mol% of Pd catalyst in 66-99% yields and exhibit good functional group compatibility. PMID- 10804565 TI - A new selective synthesis of the Ile-allo-Thr-Gly tripeptide fragment of lysobactin. AB - [formula: see text] trans-Aziridine-2-carboxylic acid derivatives are useful intermediates for the synthesis of threonine or allo-threonine through ring expansion and SN2 displacement, respectively. We describe here the preparation of the Ile-allo-Thr-Gly 11 fragment of Lysobactin via the aziridine 9 intermediate. PMID- 10804566 TI - Palladium-catalyzed coupling of vinylogous amides with aryl halides: applications to the synthesis of heterocycles AB - [formula: see text] Described herein is the first example of the palladium catalyzed coupling of vinylogous amides with aryl bromides and chlorides. The scope of this reaction with respect to the aryl component is investigated. Additionally, a tandem reaction sequence in which the above coupling is followed by an intramolecular Heck reaction is presented. These reactions can be applied to high-yielding, one-pot syntheses of nitrogen-containing heterocycles. PMID- 10804568 TI - Dihydro AB - [formula: see text] Treatment of o-propargylaryl nitrones with base provided 1,2 dihydro[c]benzazepin-3-ones in good yields. The straightforward transformation is explained on the basis of a multistep rearrangement involving conjugated allene nitrones as precursors of a 1,7-dipolar electrocyclization process that is followed by further bond reorganizations. PMID- 10804567 TI - An efficient synthesis of cyclodextrin-based carbohydrate cluster compounds. AB - [formula: see text] The photoaddition of the thiol 2,3,4,6-tetra-O-acetyl-beta-D 1-thioglucopyranose to the allyl ether functions of per-2-allyl-, per-6-allyl-, and per-2,6-diallyl-beta-cyclodextrin derivatives provides a remarkably simple and efficient way for attaching glucopyranose units onto (1) the secondary face, as well as (2) the primary face, of beta-cyclodextrin--not to mention (3) both the primary and secondary faces, simultaneously--in yields of up to 70%. PMID- 10804569 TI - Synthesis of analogues of (-)-cytisine for in vivo studies of nicotinic receptors using positron emission tomography. AB - [formula: see text] 9-Substituted analogues of (-)-cytisine were synthesized in high yields via palladium-mediated couplings of either 9-(-)-bromocytisine and organostannanes or 9-(-)-trimethylstannylcytisine and fluorobromobenzene. The protection of the amine with a nitroso group and the use of PdCl2(PPh3)2 to carry out the Stille reaction allowed the rapid synthesis of 9-(4' [18F]fluorophenyl)cytisine (18F: t1/2 = 109.7 min), a new promising radioligand (radiochemical yield: 10% from [18F]KF, 150 min, four steps) for positron emission tomography studies of alpha 4 beta 2 nicotinic receptors. PMID- 10804570 TI - First chelated chiral N-heterocyclic bis-carbene complexes AB - [formula: see text] First chiral bidentate bis-carbene complex of Pd(II) prepared from a bis-imidazolium salt derived from (S)-2,2'-bromomethyl-1,1'-binaphthyl exists in both cis- and trans-square planar geometry. These and the corresponding trans-Ni(II) complexes are remarkably resistant to high temperature, air, water, and silica gel chromatography. The Pd complexes catalyze Heck reactions. PMID- 10804571 TI - Effect of fluoride salts on metal-mediated reactions. Aluminum/fluoride salt mediated reduction and pinacol coupling of carbonyl compounds in aqueous media AB - [formula: see text] Metal fluoride salts were found to activate aluminum in water to react with carbonyl compounds to give the pinacol coupling products and/or the reduced alchohols. The metal ion of the fluoride salt was found to play a role in controlling the chemoselectivity and stereoselectivity of the reaction. PMID- 10804572 TI - Direct transformation of 1,3-dihalides into dithianes and dithiepines via a novel one-pot reaction with carbon disulfide and sodium borohydride AB - [formula: see text] 1,3-Dithianes and -dithiepines are prepared via an experimentally simple and efficient direct transformation of 1,n-alkyl dihalides utilizing carbon disulfide and sodium borohydride. PMID- 10804573 TI - Total synthesis of spicamycin amino nucleoside. AB - [formula: see text] The first total synthesis of spicamycin amino nucleoside 2 has been achieved. The aminoheptose unit 5 was prepared stereoselectively from myo-inositol, and the characteristic N-glycoside linkage was constructed by way of Pd-catalyzed coupling reaction of 5 with 6-chloropurine derivative 6. PMID- 10804574 TI - 1,4-dioxene in organic synthesis: rapid access to the oxabicyclo AB - [formula: see text] Oxidation of 2,3-disubstituted 1,4-dioxenes 3 with m chloroperbenzoic acid in methanol followed by nucleophilic addition of allyltrimethylsilane in the presence of TiCl4 afforded dienes 5, which have been converted to oxabicyclo[4.2.1] nonenes 8 in excellent yield by olefin ring closing metathesis reaction. PMID- 10804575 TI - Dirhodium(II) tetrakis[methyl 2-oxaazetidine-4-carboxylate]: a chiral dirhodium(II) carboxamidate of exceptional reactivity and selectivity. AB - [formula: see text] A new chiral azetidinone-carboxylate ligand for dirhodium(II) catalysis enhances reactivity toward diazo decomposition and selectivity toward cyclopropanation enabling diazomalonates, vinyldiazoacetates, and aryldiazoacetates to be effectively used with a dirhodium(II) carboxamidate catalyst. PMID- 10804576 TI - Novel "reverse Kahne-type glycosylation": access to O-, N-, and C-linked epipodophyllotoxin conjugates. AB - [formula: see text] Exposure of epipodophyllotoxin C4-sulfoxides to triflic anhydride, followed by a silyl glycoside, provides a glycoconjugate of the etoposide variety via formal "reverse Kahne glycosylation." To our knowledge, this is the first example of this variant of the Kahne activation method wherein the activating functionality is positioned on the aglycon, rather than on the sugar. Phenols, anilines, or allyl silanes are also efficiently captured at C4, producing the corresponding O-, N-, and C-linked lignan conjugates. PMID- 10804577 TI - An expeditious synthesis of (+/-)-desepoxy-4,5-didehydromethylenomycin a methyl ester AB - [formula: see text] A total synthesis of the racemic methyl ester of desepoxy-4,5 didehydromethylenomycin A has been achieved in six steps with an overall yield of 31% starting from diethyl methanephosphonate. The key steps include the Nazarov cyclization of the dienone 7 leading to the alpha-phosphoryl cyclopentenone 8 and the Horner-Wittig reaction of the latter employed for the introduction of the exocyclic methylene moiety. PMID- 10804578 TI - Enantioselective formal total synthesis of roseophilin. AB - [formula: see text] An enantioselective formal total synthesis of roseophilin 3 is presented. The 13-membered ring of macrotricycle 1 was formed via an efficient ring-closing metathesis reaction of bicycle 4. A palladium-catalyzed methoxycarbonylation reaction of enol triflate 5 was utilized to functionalize the right-hand ring of bicycle 2. The allyl substituent was introduced by a radical allylation of alpha-bromoketone 17. PMID- 10804579 TI - Highly stereoselective formal AB - [formula: see text] Highly stereoselective formal [3 + 3] cycloaddition reactions of chiral vinylogous amides with alpha, beta-unsaturated iminiums are described. A mechanistic model is proposed to rationalize the observed stereoselectivity. The 6 pi-electron electrocyclic ring closure appears to be reversible, and a preferred rotation of the alkenyl group, one of the three 2 pi-components, during the ring closure step provides the thermodynamically favored diastereomer as the major product. PMID- 10804581 TI - Crystal lattice photochemistry often proceeds in discrete stages. Mechanistic and exploratory organic photochemistry AB - [formula: see text] While the solid-state photochemistry of crystals has been assumed to proceed to some critical point beyond which the crystal is destroyed, it has now been found that crystal lattice photochemistry often occurs in stages or regimes. In each regime different chemistry generally results. The reaction in different stages may be followed kinetically. PMID- 10804580 TI - Synthesis of functionalized oxazolines and oxazoles with DAST and Deoxo-Fluor. AB - [formula: see text] A mild and highly efficient cyclization of beta-hydroxy amides to oxazolines is described using DAST and Deoxo-Fluor reagents. A one-pot protocol for the synthesis of oxazoles from beta-hydroxy amides is also presented. PMID- 10804582 TI - Catalytic synthesis of aldehydes and ketones under mild conditions using TEMPO/Oxone AB - [formula: see text] A novel, metal-free oxidation system for the catalytic synthesis of aldehydes and ketones--TEMPO/Oxone--has been developed. An optimized reaction protocol proved especially successful for the synthesis of ketones. Additionally, the influence of quarternary ammonium salts on the catalysis was studied. The mild conditions of this novel procedure were shown to tolerate even sensitive silyl protective groups which can otherwise be cleaved in the presence of Oxone. PMID- 10804583 TI - Jarring bodies: thoughts on the display of unusual anatomies. PMID- 10804584 TI - The origin and control of pandemic influenza. PMID- 10804585 TI - Warren McCulloch's search for the logic of the nervous system. PMID- 10804586 TI - Walter Pitts. PMID- 10804587 TI - The meme metaphor. PMID- 10804588 TI - On the significance of a "minor" phylum (the Tardigrada) in the context of a constructivist view of knowledge. PMID- 10804589 TI - The biology of signification. PMID- 10804590 TI - In search of Alexander A. Maximow: the man behind the unitarian theory of hematopoiesis. PMID- 10804591 TI - Exobiology. PMID- 10804592 TI - The stability of the genome and the genetic instability of tumors. PMID- 10804593 TI - Prevention of heart disease in women: recommendations for management of risk factors. AB - Coronary heart disease is the leading cause of death and disability in women in the U.S. Unfortunately, efforts for primary and secondary prevention of coronary heart disease are less than optimal. The need to emphasize prevention of coronary heart disease in women in important because first events are often fatal in women. Factors that are unique in terms of their influence on risk for coronary heart disease in women include age, reproductive and hormonal status, high density lipoprotein cholesterol and triglyceride levels, and the presence of diabetes. A consistent plan to address risk factor management of each woman is the key to long term risk reduction. Nurses and nurse practitioners, working in any setting, can use the recommendations presented in this paper to help women reduce their risk for coronary heart disease. PMID- 10804594 TI - An overview of essential hypertension in Americans as a multifactorial phenomenon: interaction of biologic and environmental factors. AB - This paper reviews the current literature on essential/primary hypertension in terms of its expression as a multifactorial phenomenon. The genetic and environmental risk factors involved in expression of hypertension and their interactions are discussed. A specific mutation in epithelial sodium channels, T549M, is presented as a genetic risk factor for primary hypertension as expression of this mutation has been reported to result in hyperabsorption of sodium in homozygous individuals. T549M is used in this report to illustrate the multifactorial nature of primary hypertension. Possible interactions of T549M with environmental factors known to promote hypertension and the outcome of these interactions are discussed. Data indicates that both genetic and environmental risk factors must be considered to understand and intervene effectively with patients who have primary or essential hypertension. PMID- 10804595 TI - The emergency department experience of chest pain patients and their intention to delay care seeking for acute myocardial infarction. AB - This study investigated how patients' emergency department experience was related to their intention to delay action in response to future symptoms of acute myocardial infarction. A sample of 426 persons admitted to the emergency department with a chief complaint of chest pain and released from the emergency department were contacted by telephone. Patients were queried about their affective response to the emergency department experience, their satisfaction with emergency department staff communication, their intention to delay prompt action for acute myocardial infarction symptoms in the future, the influence of others in the decision to seek care, and medical and demographic status. The results of a mixed model linear regression analysis showed that the less education patients had (p = 0.007), the less sure they felt that going to the emergency department had been "the right thing to do" (p = 0.004), and the greater the degree of embarrassment (p = 0.0001), the greater was the intention to delay action for future symptoms of acute myocardial infarction. The results also showed that those patients who were prompted by health professionals to go to the emergency department were less likely to report intentions to delay for future symptoms (p = 0.036). It is important that emergency department staff reassure chest pain patients who are sent home that they did the right thing by coming to the emergency department for their symptoms. Providers need to be particularly sensitive to feelings of embarrassment. PMID- 10804596 TI - Using a best practice approach to prevent treatment interference in critical care. AB - Treatment interference, also known as therapy disruption, device disruption, or self extubation, is a common and especially difficult clinical problem in critical care. This paper presents creative and practical clinical innovations and relevant research findings as a "best practice approach" to prevent treatment interference in critical care settings. Key principles are presented to guide patient assessment and selection of nursing strategies. Nursing assessment parameters are described and a wide range of nonrestraint strategies are discussed. PMID- 10804597 TI - Considerations for medication administration in the NPO patient. PMID- 10804598 TI - Heart failure clinics in Europe. PMID- 10804599 TI - What life threatening complication of hyperthyroidism results in prominent U waves on the EKG? PMID- 10804600 TI - The value of electron beam computed tomography in identifying high risk patients for CAD. PMID- 10804601 TI - [Quality assessment of multiple trauma management bu ISS, TRISS or ASCOT?]. AB - OBJECTIVES: Scores have been developed to assess the quality of trauma treatment. Our objective was to investigate prospectively whether there is a significant difference between the predicted survival and mortality rates with ISS, TRISS and ASCOT and observed rates in our patients. If the answer is affirmative, we also wished to evaluate whether the difference is significant enough to justify the greater data collection effort required for ASCOT as opposed to TRISS, and for TRISS as opposed to ISS. PATIENTS AND METHODS: Charts from 470 of 484 blunt multiple trauma patients with an ISS of 8 or more documented as part of a National Fund Study between June 1990 and June 1996 were reviewed. We compared the survival and mortality rates calculated by ISS, TRISS and ASCOT with the observed survival and death rates. RESULTS: Only the predicted survival and death rates calculated by ISS differed significantly from the observed rates. There were also significant differences between ISS and TRISS, and ISS and ASCOT, with regard to the predicted rates. CONCLUSIONS: In our unit ISS, which is simple to use, is not suitable for quality assessment. TRISS and ASCOT are of equal value. We prefer TRISS until a better, internationally recognised score is available, since it involves less effort and is less error-prone. General recommendations for Switzerland cannot be made until further results from other national accident and emergency units are available. PMID- 10804602 TI - [Fever of unknown origin as a sign of calcium pyrophosphate deposition disease]. AB - Calcium pyrophosphate dihydrate (CPPD) crystal deposition disease may manifest clinically as septic fever (40 degrees C), acute pseudogout attack of knee, wrist and shoulders, or as a variety of patterns of chronic inflammatory or degenerative joint disease. The association of pseudogout with fever is less widely recognised and may lead to over-investigation, delay in appropriate treatment and disproportionate costs. We report on a 67-year-old woman with a history of recurrent episodes of fever and polyarthritis every 2 months for the last 3 years. Because of this she was hospitalised several times, finally with suspected culture-negative endocarditis, and was treated for 6 weeks with gentamicin, rifampicin and vancomycin. During this therapy the patient again developed septic fever and acute arthritis of the right wrist. Radiographs of the wrist, knee and symphysis pubis revealed prominent chondrocalcinosis and destructive arthropathy. PMID- 10804603 TI - [Autologous transplantation of peripheral blood stem cells in the framework of treatment for hematologic neoplasms. II. Determination of CD34+ cells and progenitor colonies]. AB - 119 collections of peripheral blood stem cells were performed in 60 patients suffering from various haematological malignancies. One patient was transplanted twice. Peripheral blood stem cell mobilisation was performed using various regimens. A correlation (r = 0.732) was found between the number of peripheral CD34+ cells and the number of CD34+ cells that were collected by apheresis. Furthermore, the number of collected peripheral blood stem cells correlated with the number of colony-forming units (CFU-GM) identified after cell culture (r = 0.607). After transplantation, the duration of neutropenia (> 0.5 x 10(9)/l) was 11.7 days (SD = 3.3) and of thrombocytopenia (> 20 x 10(9)/l) 12.7 days (SD = 6.8). For the whole group, no correlation was observed between the number of CD34+ cells infused and the length of the aplasia. However, when the patients were separated into two groups, according to the amount of CD34+ cells transplanted (< 10 x 10(6)/kg, n = 48 and > 10 x 10(6)/kg, n = 13), a statistically significant but moderate decrease in the duration of neutropenia and thrombocytopenia was observed in patients who received the highest doses of CD34+ cells. Our results confirm the utility of measuring the number of the CD34+ cells in peripheral blood as well as in the product collected by apheresis, but they challenge the place of cultures of progenitor haematopoietic cells. Moreover, the infusion of large amounts of CD34+ cells shortens the duration of the aplasia. PMID- 10804604 TI - [Indications for the use of human albumin solutions: an expert report. Human albumin expert group]. PMID- 10804605 TI - [Vaquez disease: an unusual presentation]. PMID- 10804606 TI - [Etiology of coronary artery disease]. PMID- 10804607 TI - [Etiology of stroke--with special reference to cerebral infarction]. PMID- 10804608 TI - [Diagnosis of coronary artery diseases]. PMID- 10804609 TI - [Diagnosis of stroke]. PMID- 10804610 TI - [Therapy and prognosis of coronary artery disease]. PMID- 10804611 TI - [Therapy and prognosis of stroke]. PMID- 10804612 TI - [Prevention of coronary artery disease]. PMID- 10804613 TI - [Prevention of stroke]. PMID- 10804614 TI - [Current status of liver transplantation]. PMID- 10804615 TI - [Fulminant hepatitis: update]. PMID- 10804616 TI - [Endoscopic treatment of digestive system cancer]. PMID- 10804617 TI - [Cerebrovascular disorders--current diagnosis and treatment]. PMID- 10804618 TI - [Anorexia nervosa]. PMID- 10804619 TI - [Body fluid control: recent progress]. PMID- 10804621 TI - [Neuromuscular diseases associated with malignant tumors--with special reference to nervous system symptoms]. PMID- 10804620 TI - [Organ involvement in systemic lupus erythematosus and its treatment]. PMID- 10804622 TI - [Clinical aspects of ANCA-related diseases]. PMID- 10804623 TI - [Sudden death and arrhythmia]. PMID- 10804625 TI - [Diagnosis and treatment of diabetic nephropathy]. PMID- 10804624 TI - [Therapy of arrhythmia]. PMID- 10804626 TI - [Diagnostic process for hematuria]. PMID- 10804627 TI - [Diagnosis and treatment of malignant lymphoma]. PMID- 10804628 TI - [Clinical aspect of myelodysplastic syndromes (MDS)]. PMID- 10804629 TI - [Current form of pulmonary tuberculosis]. PMID- 10804630 TI - [Treatment of lung cancer: recent progress]. PMID- 10804631 TI - [Infections--current topics]. PMID- 10804632 TI - [Helicobacter pylori and cardiovascular pathology. Again the inflammatory hypothesis of arteriosclerosis]. PMID- 10804633 TI - [The professional burnout syndrome in resident physicians in hospital medical specialties]. AB - BACKGROUND: To evaluate the existence of burnout among medicine residents of hospitality specialties, and the relationship with sundry factors. METHODS: A transversal study was made among internal medicine residents of La Fe Hospital with more than one year in their job. For this purpose two questionnaires were used: a) the Maslach Burnout Inventory and a general questionnaire about social, demographic, residency, laborer and economic factors. RESULTS: Fifty-six (81.1%) internal medicine residents participated. Comparing with Spanish sample established with medical specialists, we found 7.1% or the residents with high scores on emotional exhaustion, while 17.8% scored high on depersonalization and 23.2% scored low on personal accomplishment. But the levels were lower on residents sample than in the medical specialties sample. Factors associated with burnout were social and demographic compass (be single), like on residency compass (the chosen specialty was not the first option), like properly laborer (low laborer satisfaction, to feel low recognition from boss or patients). We didn't find relationship with economic factors. CONCLUSIONS: There are hospitality internal medicine residents who suffer burnout and exist relationship with social, demographic, residency and laborer factors. PMID- 10804634 TI - [Lipodystrophy and hyperglycemia produced by protease inhibitors]. AB - BACKGROUND: HIV-1 protease inhibitors (PI) have been recently associated with a syndrome of anomalous body fat distribution or lipodystrophy (LD), which sometimes is associated to hyperglycemia. Many aspects of this syndrome remain obscure. The objective of the study is to describe the prevalence, clinical patterns, derangement of glucose metabolism associated and follow-up. METHODS: Patients taking at least one PI during January 1998 were asked a questionnaire about any body change, and a physical exam looking for fat changes was performed. In patients found to have LD, basal plasma glucose, cholesterol, triglycerides and cortisol were studied, and an oral glucose tolerance test (OGTT) was performed. In those cases with breast enlargement, prolactin and estradiol were measured. RESULTS: Among 272 patients, five cases (1.8%) of LD were detected. The syndrome appeared associated with all the IP available at the time of the study, a medium of 4.2 months after the introduction of the drug. The most common pattern was fat accumulation in abdomen and breast, and fat loss in arms, legs and face. Basal glycemia was normal in all 5 patients, but OGTT showed carbohydrate intolerance in two (both of them had family antecedents of type 2 diabetes mellitus). Only one patient need treatment withdrawal because of progressive deforming changes. CONCLUSIONS: The LD can appear in patients taking any IP. Body changes are usually self limited and in many patients PI don't need to be stopped. PMID- 10804635 TI - [The clinical history and physical examination in patients with chronic lumbar pain. Patient classification by a decision tree]. AB - AIM: To analyze the correlation between clinical symptoms and physical examination signs in diagnostic groups of low back pain patients; making a spanning tree. MATERIAL AND METHOD: A prospective longitudinal study of 485 low back pain patients was carried. We assess the type of pain and its irradiation, evolution in time, duration of the episodes, timetable, intensity and morning stiffness. A complete physical examination with specific tests for sacroiliac joints and neurologic examination was carried. We carried out a study of absolute and relative frequencies; calculating the rates with the chi 2 Pearson test and having a confidence of 95%. RESULTS: We found a significant relation (p < 0.0001) between the type of pain and the diagnosis groups. Proximal irradiation with functional disorders (p < 0.01) and metabolic bone diseases (p < 0.0001); the radicular irradiation and the disorders by physical agents (p < 0.001). The flare up back pain with disorders by physical agents and metabolic bone diseases (p < 0.001). The morning stiffness was exclusive in the inflammatory diseases. In the physical examination, was found a significant relation (p < 0.0001) between the different tests with the diagnostic groups. CONCLUSION: The history and the physical examination provide us symptoms and signs which let us direct towards the above diagnostic. The type of pain with its characteristics and the referred pain as a main symptoms, give us some information, which complemented with the physical examination tests, providing the base on which we support our diagnostic hypothesis, allow us the first classification of the patients; at the same time we understand as a low back pain patients suffer its problem. PMID- 10804636 TI - [Long-term portal hypertension in the rat by means of triple stenotic ligation of the portal vein]. AB - OBJECTIVE: To improve the technique of stenosing ligation of the portal vein, frequently used to obtain a prehepatic portal hypertension model in the rat, by increase the initial resistance to the blood flow. METHODS: In this experimental work portal stenosis by three equidistant stenosing ligatures which are fixed in a plastic guide is proposed as an alternative technique. RESULTS: Triple stenosing ligation of the portal vein does not produce mortality and causes splenomegaly as well as a larger degree of collateral venous circulation, particularly of splenorenal type, in relation to rats with a simple stenosing ligation. CONCLUSION: Increase of the stenosed tract length of the portal vein in the rats with triple ligation increases the initial resistance to the portal flow and then the posterior evolution would be impaired and this mechanism could be involved in a larger degree of collateral venous circulation. PMID- 10804637 TI - [Massive hemorrhage due to esophageal varices as a form of presentation of autoimmune hepatitis]. AB - Autoimmune hepatitis is a necro-inflammatory disease of unknown etiology. An acute or fulminant presentation is rare. We report a 18-year-old man presented with acute variceal hemorrhage. The patient was treated with endoscopic sclerotherapy, somatostatin, balloon tamponade and TIPS. The patient died within a week of massive bleeding and hepatic failure. This patient's history strongly suggests that autoimmune hepatitis can be presented as fulminant hemorrhage. The importance of diagnosis and treatment is emphasized to improve prognosis. PMID- 10804638 TI - [Disseminated intravascular coagulation in a patient with polyarteritis nodosa and bilateral hypoacusis]. AB - We describe the case of a 44-years-old female patient with a panarteritis nodosa preceded by a rapid progressive bilateral deafness after an acute otitis media. Once on treatment with steroids and cyclophosphamide, she developed a disseminated intravascular coagulation, with a fatal evolution not justifiable by a different cause apart from her main disease. PMID- 10804639 TI - [A case report of mixed, parenchymatous and meningo-basal neurocysticercosis]. AB - Neurocysticercosis is the most common parasitic disease of the central nervous system. Its incidence diminished dramatically in the last decades in our country, but due to emigration from endemic areas is expected an increase in the next years. In spite of the new immunologic and neuroradiographic techniques, diagnostic of centeinty may be very difficult. In this way exists controversy about the appropriate treatment. We present a case of mixed neurocysticercosis with parenchymal and meningeal disease that shows the difficulty of this disease in its diagnostic and treatment. PMID- 10804640 TI - [Acute polyarthritis associated with parvovirus B19 infection]. AB - We describe one female with acute polyarthritis and transitory rash. The arthritis of sudden onset, is localized in large and small joints and accompanied by significant stiffness, particularly noted in early morning. The symptoms only improvement a few with nonsteroidal anti-inflammatory drugs and the arthritis resolved within a few days. The diagnosis is based in the presence of anti-B19 IgM. The differential diagnosis of the acute polyarthritis is extensive, serological test should be made when we suspect the presence of a viral arthritis. PMID- 10804641 TI - [Congestive heart failure due to diastolic dysfunction]. PMID- 10804642 TI - [Panhypopituitarism associated with an empty sella turcica]. PMID- 10804643 TI - [Thrombotic thrombocytopenic purpura associated with HIV]. PMID- 10804644 TI - [Bone metastasis, the first manifestation of hepatocarcinoma]. PMID- 10804645 TI - [Ambiguity in a publication: is it a clinical trial or not?]. PMID- 10804646 TI - [Prostatic biopsy and spondylodiscitis]. PMID- 10804647 TI - [Cardiac tamponade during Staphylococcus aureus sepsis in an AIDS patient]. PMID- 10804648 TI - [Liver toxicity from heparin]. PMID- 10804649 TI - [Interferon treatment in chronic HCV hepatitis and autoimmune hypothyroidism]. PMID- 10804650 TI - [Community-acquired bacteremic pneumonia due to Citrobacter diversus]. PMID- 10804651 TI - [Nonchromaffin paraganglioma of the retroperitoneum and non-Hodgkin lymphoma, a rare association]. PMID- 10804652 TI - [Prolonged fever in a patient with hepatic space-occupying lesions of long evolution]. PMID- 10804653 TI - [Retropneumoperitoneum as the manifestation of an atypical diverticulitis of the sigmoid]. PMID- 10804654 TI - [The content and limits of consent to a medical procedure]. AB - The bases of legality for any medical-surgical activity aimed at a therapeutical diagnostic purpose is the consent of the patient. Such a consent must comply with some requirements of validity, first of all the owner of this right must be able to realize the significance of the assertion of his will. Besides, the awareness of the consent assumes that the surgeon should make a complete and detailed planning of the operation, with a clear reference to any possible hazard, as well as to any possible after-effects, as far as either the intellectual level or the motivity of the patient are concerned. After having examined the various laws covering, the legal, doctrinal and jurisprudential views, authors defines the aspects of the therapeutical treatment. The differences between a free decision and a true informed consent are outlined as well as the extension and the expression of same. It is particularly analysed the relationship between capacity and competence to get the juridical right. The question of understanding which is the best way to respect individual freedom, often reduced to a virtual level form a pathological restriction of individual autonomy, is still open. Moreover, the consent may really be effective only if it is specific, the is aimed at a well definite and accurate therapeutical act. Authors agrees with the criminalist interpretation that the medical surgical activity, out of consent of the patient, can give form to personal voluntary injury. In case of medical surgical treatment different from that previously agreed and out of a situation of therapeutic flagrant necessity, the authors are of the opinion that the inquiry has to concern the medical behaviour in order to estimate the medical surgical treatment as personal voluntary injury or as personal negligent injury. Some reasoned evaluations on the problem were considered at the end of the article. PMID- 10804655 TI - The new international staging system for lung cancer. Evolution of the system and concerned remarks. AB - The rationale of the Staging System of Lung Cancer is discussed from his presentation (Mountain, 1985) to the recent revision and proposals of new classifications. Survival rates offered a strong statistical support to the latest revision in 1997. Stage Group have become 7 out of Stage 0 (Tis). In the New Lymph Node Map, station 4 is confirmed as mediastinal (N2). The improved definition of Stage Grouping requires a golden standard of staging and a worldwide consensus on the surgical approach to mediastinal lymphadenectomy. IASLC, the International Association for the Study of Lung Cancer, is now moving to collect a new largest database with the aim to offer the next expected Revision. PMID- 10804656 TI - The "Will Rogers effect" on stage grading. AB - Will Rogers phenomenon affects survival statistics applied to clinical research and could determine a misreading of results. Stage migration due to new methods of diagnostic imaging and staging invasive procedures could improve actuarial survival in each stage. TNM System is impaired when survival rates come from different inhomogeneous countries, regions and eras. Randomized trials suffer this fallacious phenomenon when staging depends on the different treatments which are to be evaluated. PMID- 10804657 TI - Circulating biomarkers for lung cancer. AB - Ideal characteristics of tumor markers and their role in clinical practice are discussed. Measuring markers in screening programs resulted inefficient, while the pathological elevation of one or more markers may be virtually diagnostic of lung cancer in appropriate clinical setting. The elevation of CEA, NSE, TPA and other presented, appears particularly useful in the evaluation of the extent of disease, and may correlate with prognosis, as well as in postsurgical follow-up and monitoring therapy. PMID- 10804658 TI - Detection of disseminated lung cancer cells in lymph nodes by monoclonal antibodies: impact on staging and prognosis. AB - The recent development of monoclonal antibodies has made possible the detection of tumoral cells in blood and bone marrow. The immunohistochemical assay allows the discovery of metastatic cells in lymph nodes of patients with NSCLC. Using the monoclonal antibody Ber-Ep4, positive cells were demonstrated in 21.6% of 125 patients staged as tumor free by conventional histopathology. No correlation was detected between Ber-Ep4 positive cells and other pathological factor (T factor, Histology, Grading). Positive nodal cells were associated with a reduced disease free survival. This test could represent a new criterion for adjuvant therapy and for selecting high-risk patients. PMID- 10804659 TI - Radiological assessment of clinical staging of lung cancer. AB - Technical details of volumetric spiral CT and high resolution CT are presented. The role of CT scan in lung cancer is discussed: confirmation of a suspected lesion, identification of an unknown one, clinical staging, planning bioptic procedures and follow-up. In clinical staging, CT scan measures tumor diameter and relationship with surrounding structures (T factor) as well as investigates about nodal status (N factor) at the hilum or in the mediastinum but the limitation is due to the difficulty of distinguish between nodal inflammatory enlargement and metastatic involvement. Moreover, CT can be extended to the upper abdomen aimed of assessing adrenals, kidneys and liver (M factor). PMID- 10804660 TI - The value of transesophageal echography in the clinical staging of lung cancer. AB - The noninvasive preoperative assessment of lung cancer is currently based on CT scan. This technique is used to find out the local extension of the tumor and to identify the presence of mediastinal adenopathies by distinguishing as accurate as possible between benign and metastatic adenopathies according to their size. When CT scan suggests the presence of suspected mediastinal lymph nodes, invasive diagnostic methods are indicated to confirm whether the lymph nodes are affected (transbronchial needle aspiration, mediastinoscopy, mediastinotomy and thoracoscopy). Transesophageal echography (TEE) was first used in the 80's as a diagnostic guide in the clinical classification of gastrointestinal tumors and may be of help in the mediastinal evaluation of lung cancer as well. Several studies have demonstrated the value of TEE in detecting mediastinal adenopathies and mediastinal structures involvement (great vessels and mediastinal organs) by assessing the direct growth of the primary tumor which is crucial in the surgical decision. To distinguish benign from metastatic nodes TEE, unlike CT scan, does not consider their size only but other issues as shape, definition of edges and echographic structure. Generally speaking, TEE detects more adenopathies than CT scan but not in all mediastinal stations since it is limited by the presence of air; thus, adenopathies located in the upper mediastinum, mainly on the right side, are usually missed by TEE. The areas more clearly visualized are the subcarinal, paraesophageal, tracheobronchial and hilar ones and mainly those located in the aortopulmonary window. Currently, TEE can be used also to guide a thin needle toward the lymph nodes and to perform its aspiration biopsy. In conclusion, transesophageal and/or endobronchial echography are developing promising diagnostic methods which provide additional information that can influence the clinical decision making process and the prognosis. They are well tolerated by the patients but time is needed to find out their role. Lastly, their usefulness will depend also on their feasibility since an expensive equipment and specifically well trained staff are needed. PMID- 10804661 TI - Integration of TBNA and TCNA in the diagnosis of peripheral lung nodules. Influence on staging. AB - Although transbronchial and percutaneous approaches to peripheral pulmonary lesions (PPL) are widely diffused, effective and safe diagnostic techniques, no standardised strategy yet exists that defines the specific role of each in this clinical setting. With the aim of defining the role of each approach and of verifying if the two techniques are alternative or may be integrated in a logical diagnostic sequence based on the advantages and the limits of each biopsy method, the authors analysed their experience with the integrated use of transbronchial and percutaneous approach to PPL performed on 1,680 consecutive patients affected by PPL. The procedure used was as follows: 1) bronchoscopy with exploration of the bronchial tree and transbronchial needle aspiration (TBNA) of hilar/mediastinal lympho nodes for staging N factor followed by TBNA and transbronchial pulmonary biopsy (TBPB) of the PPL under fluoroscopic guidance and immediate cytological assessment (ICA); 2) if TBNA was diagnostic, the procedure was stopped; if not, a second pass with the needle was performed and then the bronchoscope removed; 3) if the second TBNA was not diagnostic, percutaneous approach (PCNA) with ICA was performed up to a maximum of three needle passes. Diagnostic sensitivity for malignant lesions was: 50.0% for TBPB, 70.1% for TBNA, 76.0% for TBPB and TBNA together, 92.8% for PCNA, and 95.0% overall. The percentage of benign nodules correctly defined was 43.0% for TBPB, 16.7% for TBNA, 47.8% for PCNA. Examination of the upper airways and bronchial tree was positive for lesions endoscopically visible in 11.4% of cases. TBNA of hilar/mediastinal lympho nodes was positive for metastatic involvement in 43.9%. The authors' experience demonstrates that transbronchial and percutaneous approaches must be considered complementary and that their integrated use not only increases diagnostic yield but also permits important information to be obtained for the staging of lung cancer. The creation of teams able to utilise both approaches with the cytopathologist present for ICA should be encouraged in order to optimise the diagnostic management of PPL. PMID- 10804662 TI - Anatomical pathways of lymphatic flow between lung and mediastinum. AB - The knowledge of anatomical lymphatic drainage pathways between lung and mediastinum and metastatic spread patterns of lung cancer forms the basis for the modern TNM classification for the staging of lung cancer. Clinical and anatomical studies divided pulmonary nodes into intrapulmonary and bronchopulmonary, the latter distinguished into lobar and hilar nodes. Mediastinal nodes are grouped into anterior prevascular, tracheobronchial, paratracheal and posterior nodes. The different pathways of lymphatic drainage of the lungs to the mediastinal lymph nodes are presented with the consequent surgical implications. PMID- 10804663 TI - Surgical exploration of the mediastinum by mediastinoscopy, parasternal mediastinotomy and remediastinoscopy: indications, technique and complications. AB - Imaging diagnostics often fail to provide enough certainly to make therapeutic decisions, since radiological images do not always correlate well with the pathological condition of the lesions. Surgical exploration of the mediastinum by mediastinoscopy allows to obtain very accurate information from inspection, palpation and biopsies of lymph nodes or tumors directly affecting the mediastinum. Mediastinoscopy assesses the upper mediastinum, including nodal stations 1, 2R, 2L, 3, 4L, 7, 10R and 10L. It can also assess direct invasion of the mediastinum from adjacent tumors. Parasternal mediastinoscopy is a complementary technique to reach nodal stations 5 and 6, which cannot be reached with standard cervical mediastinoscopy. Remediastinoscopy has been performed to restage tumors after delayed treatment and to stage second primary and recurrent tumors. It has proved useful, too, to restage N2 lung cancer after induction chemotherapy. In all these indications, remediastinoscopy was technically possible. All these techniques are associated with very few complications (around 3%) and a low mortality rate of less than 1%. PMID- 10804664 TI - Clinical value of cervical mediastinoscopy in the staging of bronchial carcinoma. AB - Whether mediastinoscopy is indicated in the preoperative staging of bronchogenic carcinoma is still a controversial issue. It may be performed routinely (to exclude locally inoperable patients from surgery), selectively, or it may be regarded as superfluous (in centers which prefer an extended lympho adenectomy at the time of thoracotomy). We regard mediastinoscopy as indicated for the following purposes: 1) staging of NSCLC and SCLC; 2) diagnostic (mediastinal masses or lung tumors without previous histology); 3) restaging after primary chemotherapy; 4) assessment of prognosis in patients with borderline operability. The indication for 224 mediastinoscopies performed at our institution in the period from September 1991 through March 1999 was mainly for staging (59.2%) or diagnostic (30.6%). Eight (5.4%) patients underwent mediastinoscopy for the assessment of operability, and 7 (4.8%) after primary chemotherapy for the restaging of loco-regionally advanced lung cancer. Sensitivity and specificity rates were 87% and 100%, respectively, with an accuracy of 93% for the mediastinoscopy performed for the staging of lung cancer at all stages. If we consider the N2 tumors (42 cases) alone, the sensitivity was 76.7% and the specificity 100%, with an accuracy of 83.3%. Overall positive and negative predictive value resulted 100% and 87%, respectively, according to the data reported in literature. Our data confirm the role of mediastinoscopy as the gold standard for regional staging of lung cancer. PMID- 10804665 TI - The invasive staging and the role of complete resection in the surgical treatment of NSCLC. AB - Years of debates couldn't solve the discussion between the NSCLC assessment founded on CT scan and mediastinoscopy as in the Western countries and the refined extensive bronchoscopy, CT imaging and exploratory thoracotomy as practiced in Japan. Recently, the clinical onset of combined therapy protocols, the recognised value of the intrathoracic staging (also in the West) and survival rates in the earlier N2 disease moved towards change this steady situation. The role of complete resection in N2 NSCLC is therefore debated from the preoperative assessment to survival results in resected cases. Accuracy of CT scan and cervical mediastinoscopy is discussed also in the light of neoadjuvant therapy. The clinical value of intrathoracic staging is improved by Japanese experiences while a rationale assessment of Complete/Incomplete Resections is defined. Moreover, technical details of intraoperative recognition are cleared. PMID- 10804666 TI - The complete hilar-mediastinal lymph node dissection. AB - Starting from Cahan's "radical pneumonectomy" and "radical lobectomy", mediastinal lymph node dissection was introduced in Japan by Ishikawa and survival results analyzed by Naruke. Japanese Lung Cancer Society (JLCS) introduced Naruke's lymph node map to standardizing dissection. Upper mediastinum, subcarinal, interlobar and upper lobar lymph nodes are to be dissected for tumors located in the right upper lobe, and the same areas in the case of middle lobe. In tumors of the lower right lobe, also nodes of the lower mediastinum should be dissected. When the tumor is in the left upper lobe, upper mediastinum (except pre and paratracheal lymph nodes), subcarinal, interlobar and lobar nodes should be dissected. Finally, in left lower tumors, lower mediastinum is to be dissected. Super-radical dissection is performed through a median sternotomy to reach pre and paratracheal nodes in tumors affecting the left upper lobe. PMID- 10804667 TI - Tumor size and extension of lymph node metastases in N2 lung cancer. AB - In all patients with NSCLC, systematic nodal dissection was performed. Since 1981, 218 stage IIIA-N2 cases were resected. We registered 2 operative mortalities. Overall survival was 23% while, in completely resected cases, survival amounted to 30%. For long-term survival, favourable prognostic factors were cN2 T1-2 N2M0, single mediastinal node involvement and a tumor 20 mm or less in maximum size. The 5 yr survival rates of stage IIIA N2 patients was, respectively 48.1% with tumor diameter < 20 mm, 27.7% with diameter between 21-30 mm, 31.2% with diameter between 31-50 mm and, finally, 16.7% with tumors larger than 51 mm. When micrometastases to lymph nodes in p-stage I (stained with H-E) were examined with immunohistochemical staining, 27% (36 patients of 132) showed micrometastases. PMID- 10804669 TI - Induction therapy in non-small-cell lung cancer: a comparison of clinical and post-surgical staging. AB - In the last decade, several neoadjuvant trials for NSCLC patients with mediastinal lymph node involvement (N2) have been scheduled. The uniform plan is based on clinical staging, therapy, clinical re-staging, surgery (when is possible) and, finally, pathological staging. The precise classification of tumor during the three different staging procedures is mandatory. Considering clinical re-staging and pathological staging, nowadays surgery could be considered correct for most of the patients enrolled in the neoadjuvant protocols including cases where a major clinical response has not been achieved. Several experiences demonstrated how often the clinical restaging overesteems neoplastic tissue by fibrosis and scar and could judge as unresectable patients with a minimal residual disease. PMID- 10804668 TI - Considerations about tumor size as a factor of prognosis in NSCLC. AB - A literature review of the initial attempt to correlate tumor size in NSCLC with the expectancy of survival is presented starting from the 60s. The larger size was connected with an increased risk of metastatic diffusion. In the 70s resulted evident the relationship between tumor size and lymph node involvement so affecting survival. In the context of the TNM Staging System (Mountain 1986) size appeared a well assessed factor of prognosis and is recognised to play a major role in Stage I where the subsets T1N0 and T2N0 showed a consistent difference in survival across the 3 cm cut-off. The peculiar relation between largest size and mediastinal lymph node metastases is discussed as well as the proposal to allocate T2 descriptor within the range 3-5 cm. Finally, series of clinical observations from Japanese experience about small sized T1N0 tumors are presented and discussed. PMID- 10804670 TI - The Japanese database for the staging of lung cancer. AB - From the beginning of the 60s, Japanese Lung Cancer Society (JLCS) proposed an original TNM classification of lung cancer based on a database of resected cases. The final step was the UICC classification edited in 1973. Complete hilar mediastinal lymph node dissection was started in the same era and the concept of station number of lymph nodes was introduced. The proposal of definition of "early lung cancer" came from Japan with the subgroups of hilar and peripheral type. A new national database is now collected for the next revision of UICC TNM classification to be edited in 2007. PMID- 10804671 TI - The Spanish data base for the staging of lung cancer. Experience of the Bronchogenic Carcinoma Cooperative Group of the Spanish Society of Pneumology and Thoracic Surgery (GCCB-S). AB - The Bronchogenic Carcinoma Cooperative Group of the Spanish Society of Pneumology and Thoracic Surgery (GCCB-S) has been enrolled in a prospective data collection project from patients with bronchogenic carcinoma who underwent thoracotomy in 20 Spanish hospitals from October 1993 to September 1997. In this 4-year period, 2995 patients were registered and demographic, clinical, biological, diagnostic, surgical, pathological and follow-up information collected in a homogeneous way. The main objectives of the Group are to describe the population with lung cancer at the time of diagnosis in Spain; to analyse operative morbi-mortality; and to identify multiple prognostic factors. So far, some preliminary results have already been published concerning the description of the population, the methodology of the Group, the assessment of morbi-mortality, and the validation of the 1997 TNM classification. PMID- 10804672 TI - Clinical experience in staging of lung cancer at Martin-Luther University Halle Wittenberg. AB - Staging of lung cancer needs an accurate diagnostic programme resulting in therapeutic and prognostic consequences. A modern, articulate flow-chart is presented and discussed. As a result, the rate of exploratory thoracotomy was reduced from 15.1% in 1988 to 2.1% in 1997 and the rate of resectability raised up to 30%. Preoperative over-staging was found in about 25% and the same value for understaging. Looking to the N-values, there was concordance of clinical and postoperative data in 61.9% of cases. PMID- 10804673 TI - [The surgical treatment of primary hyperparathyroidism: clinical experience]. AB - The primitive hyperparathyroidism (PHPT) constitutes still cause of discussion both from the diagnostic point of view and from the therapeutic one although surgical successes are generally reported. Between the most important problems there is the increase of the HPT asymptomatic or oligosymptomatic patients with the decisional difficulties in the timing of the surgical treatment and the difficult framing of the HLP disease associated with MEA and the relating surgical failures. Besides some authors support an unilateral dissection of the neck in patients with adenoma diseases diagnosed before the intervention against the traditional address of a bilateral exploration. Our experience is based on 31 patients subjected to intervention of parathyroidectomy for primitive HPT: 26 carriers of adenomas, of which 1 double, and 5 of diffused hyperplasia. We have effected 25 simple parathyroidectomy for adenoma, 1 resection of three parathyroid glands for double adenoma, 2 subtotal parathyroidectomy (7/8) for diffused hyperplasia. 2 patients had new surgical treatment for persistent hypercalcemia, and they were respectively carriers: 1 of a second ectopic adenoma and 1 of asymmetrical hyperplasia; 2 patients finally, operated in other hospitals had a second exploration and they were affected from MLP. In 26 patients we had very good results, in 2 persistence of hypercalcemia (patients who had a second look) and 3 hypocalcemia. PMID- 10804674 TI - [The surgery of abdominal aortic aneurysms synchronous with neoplastic lesions (apropos of 18 personal observations)]. AB - The management of simultaneously occurring abdominal aortic aneurysm and malignancy is controversial. It is unclear whether to treat the aneurysm first or the malignancy, or both simultaneously. If the malignancy is resected first there is a risk of postoperative rupture of the aneurysm. If simultaneous surgery is performed there is a risk of prosthetic graft infection. This condition leads to many therapeutic problems which, by the light of 18 personal cases occurred in almost ten years and the recent literature, are discussed in this paper. PMID- 10804675 TI - [Our directions for the treatment of colorectal cancer]. AB - Authors make a review of personal experience in colorectal surgery from 1998 to 1997 (18 year) and indicate the own opinion about oncological principle of colorectal surgery. Colo-rectal cancer is the most frequent tumor in West Countries (United States 156,000 new case/year, Europe 58,300 deaths/year). A Medline analysis using terms "colon and rectal cancer" prove a progressive increasing of interest about many problems regard prevention, diagnosis, prognosis and therapy of colorectal cancer. Prognosis of colorectal cancer depend on correct and complete stadiation. The most used classifications are Dukes classification, modified from Astler-Coller, and TNM classification of UICC. In the past years oncologic al principle of colorectal surgery was modified from a new debated concept of oncological radicality. The authors analyze own experience in comparison to Literature about the "no touch isolation technique", the principle of vascular ligation at the origin of vessels, lymphectomy extension, total mesorectal excision, surgical decision about advanced loco-regional cancer, surgical decision of complicated colorectal cancer (perforation and occlusion), the role of bilateral prophylactic ovariectomy, surgical treatment of liver metastasis, correct indication of T.E.M. and finally the role of laparoscopic surgery for the treatment of colorectal cancer. PMID- 10804676 TI - Paraganglioma of the bladder: report of a case. AB - Paragangliomas are uncommon tumours that comprise less than 0.1% of all primary bladder neoplasms. Approximately half of the patients present the clinical triad of hypertension, haematuria and attacks associated with micturition. About 10% of vesical paragangliomas behave malignantly. This may be manifested as local invasion or metastatic spread, particularly to lymph nodes and lung. The best therapy remains total excision and partial cystectomy is considered adequate treatment for most cases. We report a case of paraganglioma of the bladder with extramural appearance in a 39-year-old man. Clinical symptoms were hypertension, dysuria and a well-defined hypogastric mass. One year after complete removal of the tumour, the patient is asymptomatic and normotensive, with normal catecholamine and vanillylmandelic acid (VMA) levels. PMID- 10804677 TI - [Castleman disease: a clinical case report]. AB - Castleman's disease is an enigmatic lymphoid disease of unknown etiology and has different clinical manifestations. The authors present a case of a young woman with cervical localization of Castleman disease, asymptomatic. The disease was misdiagnosed and was treated for five years as thyroiditis, diagnosis confirmed by ultrasonography, scintigraphy and cytology. The particular size of the neoformation and its constant growth leaded the authors to the surgical treatment. The thyroid was normal both macroscopically and with biopsy. The neoformation was 10 cm of diameter and covered the left side of the thyroid. PMID- 10804678 TI - Anxiety in ethnic minority youth. Methodological and conceptual issues and review of the literature. AB - Although current research has documented a relatively high prevalence of anxiety disorders in American youth, this research has been conducted mainly with nonminority samples. Fair treatment and increasing numbers of ethnic minority persons in the United States require that more should be known about minority youth. However, research with majority youth cannot be safely generalized to minority youth for several reasons, such as potential differences in the manifestation of anxiety, differences in style of response to assessment devices, and different life circumstances. This review is presented in two major sections. First, the authors address definition of terms and fully examine the significance of studying anxiety in ethnic minority youth. Also considered are methodological issues such as sampling and participation biases. Second, the authors review anxiety in ethnic minority children and adolescents in the United States including studies addressing fears, worries, trait anxiety, test anxiety, and anxiety disorders. PMID- 10804679 TI - Social support in a worksite smoking intervention. A test of theoretical models. AB - To better understand how social support operates in smoking cessation, three models of support were tested: main effect, stress-buffering, and indirect. Employees from 61 worksites received (a) self-help manuals (SH); (b) self-help manuals and incentives (I); or (c) self-help manuals, incentives, and social support groups (G). At 24 months, results suggest that the main effect model of social support was operating in this study. That is, social support had a direct influence on smoking cessation. The group intervention significantly enhanced positive partner support across all time points, and partner support facilitated quitting smoking. Higher levels of appraisal support also significantly predicted successful quitting. The beneficial effects of social support were sustained throughout the process of quitting and long-term maintenance. PMID- 10804680 TI - A review of behavioral treatments used for Lesch-Nyhan syndrome. AB - Lesch-Nyhan syndrome is a genetic disorder resulting in hyperuricemia, choreoathetosis, mental retardation, and self-mutilation. The most salient feature of this disorder is the self-injurious behavior (SIB). Although the utility of behavioral interventions with SIB has been well documented, behavioral interventions with Lesch-Nyhan syndrome have been limited in number and long-term success. This article reviews the behavioral treatments that have been used in treating individuals with Lesch-Nyhan syndrome and discusses the strengths and weaknesses of these methods. Suggestions for future directions in the use of behavioral interventions for controlling SIB in Lesch-Nyhan syndrome are provided. PMID- 10804681 TI - Fear of intimacy among dating couples. AB - Fear of intimacy among heterosexual dating couples was examined with the Fear-of Intimacy Scale (FIS) and the Personal Assessment of Intimacy in Relationships (PAIR). Following a 6-month interval, couples were again contacted to determine whether they continued to date. Males reported higher FIS scores than females, and FIS scores were positively correlated within couples. Also, FIS scores of males and females were significantly correlated with indices of actual and desired intimacy; however, for females, correlations of FIS scores with desired intimacy were significantly lower than correlations with actual intimacy. Females who were no longer in the dating relationship at the 6-month follow-up had higher FIS scores than those who were continuing in their dating relationship. These findings increase our understanding of fear of intimacy, especially gender differences in dating couples. PMID- 10804682 TI - The influence of personality and history of sexual victimization in the prediction of juvenile perpetrated child molestation. AB - Structural equation modeling was used to delineate the relationship between sexual victimization and personality variables in the prediction of patterns of child molestation in adolescent males. Two hundred thirty-five adolescents, representing subsamples of sexually victimized and nonvictimized, perpetrating and nonperpetrating, and emotionally maladjusted and nonmaladjusted youths, participated in the study. Juvenile child molesters were found to have more pessimistic explanatory styles and deficits in self-sufficiency relative to nonoffending youths. A younger age at time of victimization, a greater number of incidents, a longer period of waiting to report the abuse, and a lower level of perceived family support postrevelation of the abuse were found to be predictive of subsequent sexual perpetration. Implications for treatment are discussed. PMID- 10804683 TI - The impact of interpersonal patient and therapist behavior on outcome in cognitive-behavior therapy. A review of empirical studies. AB - Empirical studies are reviewed, the aim being to investigate characteristics of the therapeutic relationship in cognitive-behavior therapy (CBT) and to identify therapist or patient interpersonal behavior that affects treatment outcome. CBT is characterized by a more active and directive stance on the part of the therapists and higher levels of emotional support than are found in insight oriented psychotherapies. Therapists express high levels of empathy and unconditional positive regard, similar to those expressed by insight-oriented psychotherapists. Two clusters of interpersonal behavior have been identified that are clearly associated with CBT outcome: (a) the Rogerian therapist variables--empathy, nonpossessive warmth, positive regard, and genuineness; and (b) therapeutic alliance. There is some evidence for the impact on outcome of three additional clusters of patient behavior: (a) the patients' perception of the therapist as being self-confident, skillful, and active; (b) the patients' openness to discuss their problems; and (c) the patients' pretreatment predisposition to change and to accept psychological treatment as a means of achieving this. It is further concluded that relationship factors in general have a consistent but moderate impact on CBT outcome. PMID- 10804684 TI - Gonorrhoea incidence in England rises again. PMID- 10804685 TI - COVER programme: October to December 1999 vaccination coverage statistics for children up to 5 years of age in the United Kingdom. PMID- 10804686 TI - [Self-training in needle-knife sphincterotomy]. AB - AIM: To analyze our experience in self-training in needle-knife sphincterotomy and to estimate the number of procedures necessary to reach competence in this technique. METHOD: Retrospective analysis of the first 60 needle-knife sphincterotomies performed, paying attention to success rates, the need for one or two attempts and the incidence and severity of early complications. The results were analyzed and divided into 2-year periods. The training period was subsequently determined using a qualitative method which combines the three above mentioned criteria and sets competence level at 90%. RESULTS: Of the 60 attempted needle-knife sphincterotomies (five in Billroth-II patients), 54 (90%) were completed. The procedure was required in 20% of all the sphincterotomies completed. Needle-knife sphincterotomy was completed at the first attempt in 39 (65%) patients and complications appeared in 13 (22%), three (5%) of which were serious but not fatal. Analysis of 2 year periods showed an almost 100% increase in the total number of needle-knife sphincterotomies completed and in those completed at the first attempt, as well as a reduction in the complication rate, which subsequently stabilized at below 15%. Forty needle-knife sphincterotomies were needed to reach competence level. CONCLUSION: Needle-knife sphincterotomy can be learned without training from an endoscopist with experience in this technique. Competence can be reached after 40 procedures. Nevertheless, we propose an alternative method to our own to achieve competence in less time and with fewer complications. PMID- 10804687 TI - [Helical CT compared to ERCP in obstructive biliary pathology]. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) is the examination of choice in the diagnosis of biliary tract pathology although newer, less invasive techniques with high diagnostic sensitivity should be evaluated. The aim of this study was to evaluate the diagnostic efficacy of helical computed tomography (CT) in bile duct obstruction, particularly in choledocholithiasis. PATIENTS AND METHODS: Forty-four patients with suspected bile duct obstruction were prospectively evaluated. Helical TC was carried out 12 hours before ERCP and patients were grouped according to risk of choledocholithiasis. RESULTS: Choledocholithiasis was found in 20 patients (45.4%). Helical TC identified this pathology in 17 and correctly ruled it out in 18 of 24 patients (S: 85%, E: 75%, positive predictive value 74%, negative predictive value 85.7%. Of the patients with choledocholithiasis, bile duct dilatation was found in 18. Helical TC correctly diagnosed this pathology in 16 of the 18 patients (88.8%). Helical TC correctly diagnosed one of two patients (50%) with choledocholithiasis and normal bile ducts. CONCLUSIONS: Helical TC is effective in the diagnosis of choledocholithiasis but is not sufficiently accurate to be used in the screening of this entity. PMID- 10804688 TI - [Cyclosporin in an oral microemulsion as the initial treatment of corticoid refractory ulcerative colitis]. AB - AIM: To assess the efficacy of oral microemulsion cyclosporin as the initial treatment of active ulcerative colitis refractory to steroid therapy. PATIENTS AND METHODS: The evolution of five patients treated with a dose of 5 mg/kg/day was analyzed. RESULTS: Oral microemulsion cyclosporin controlled acute attacks of the disease and led to remission during the 3 months of treatment in four of the five patients. Average hospital stay was 5.5 days. CONCLUSIONS: Oral microemulsion cyclosporin is an effective treatment in ulcerative colitis. Its administration is simple, the use of venous catheters is avoided and it may reduce mean hospital stay. PMID- 10804689 TI - [Autoimmune hepatitis associated with thyroiditis and hypophysitis. A case report]. AB - Autoimmune hepatitis primarily affects women and 40% of cases are associated with extrahepatic autoimmune dysfunction. Thyroiditis, ulcerative colitis and rheumatoid arthritis are the most commonly implicated entities. We present a 46 year-old woman with type-II autoimmune hepatitis and Graves disease who presented deterioration in level of consciousness, her symptoms mimicking severe liver failure. Hormone studies and nuclear magnetic resonance imaging revealed hypophysitis, which led to hypothyroidism and metabolic encephalopathy. The syndrome was resolved with hormone replacement therapy. PMID- 10804690 TI - [Mesenteric fibromatosis as the cause of a single abdominal mass]. AB - We present a mesenteric desmoid tumor in a man without associated diseases. Characteristics of this tumor are reviewed and the need to perform total exeresis of the mass for diagnosis is emphasized. Although surgery is the treatment of choice, alternative treatments are recommended when surgery cannot be performed, has been incomplete, or in cases of repeated recurrence. PMID- 10804691 TI - [Colonic balantidiasis]. AB - Balantidium coli is a Protozoa that is not usually pathogenic in man, although epidemics have been described in tropical areas. It mainly affects the colon and clinical presentation varies from asymptomatic forms to severe dysenteric syndromes. We present a case of endoscopically diagnosed colonic balantidiasis and review the most important characteristics of this parasite-induced disease. PMID- 10804692 TI - [Cerebrovascular accident as the form of presentation of a pancreatic cystadenocarcinoma: an unusual form of presentation in a rare tumor]. AB - Pancreatic cystadenocarcinoma is an uncommon neoplasm of the pancreas. Mucinous cystadenocarcinoma is the most frequent pancreatic cystadenocarcinoma. Symptoms are often non-specific but abdominal pain and jaundice are common in tumors localized in the head of the pancreas. Thromboembolic manifestations of pancreatic neoplasm have been described but are infrequent. The commonest manifestations are migratory thrombophlebitis, mesenteric venous thrombosis and pulmonary embolism. Cerebral stroke as the first manifestation of pancreatic carcinoma is exceptional. The mechanism by which pancreatic carcinoma induces stroke is unclear, although Trousseau's syndrome, complications of mucinous cancer or blood disorders could play a role. We report the case of a young woman with transient stroke and paraneoplastic thrombocytosis in whom pancreatic cystadenocarcinoma was diagnosed. PMID- 10804693 TI - [A critical review of the diagnostic methods for Helicobacter pylori infection]. PMID- 10804694 TI - [Hereditary familial amyloidosis]. PMID- 10804695 TI - [Hepatitis C: viral load and plasma lipids]. PMID- 10804696 TI - [Is it safe to drink directly from soft drink cans?]. PMID- 10804697 TI - [Anal carcinoma in Crohn disease]. PMID- 10804698 TI - [Giant lipoma of the right colon]. PMID- 10804699 TI - Gender differences in supported employment. AB - Questions about gender equity have been asked in many aspects of the disability field and have resulted in findings that women with disabilities have significantly different experiences than do men. We analyzed an existing database of information on supported employment and natural supports to ascertain whether gender plays an important role in the employment of people with mental retardation. The findings suggest that there are several important differences. Although women were perceived as being more socially appropriate on several dimensions, they worked in jobs traditionally stereotyped by gender. Women also typically worked fewer hours than did men and, therefore, earned less money, although not statistically significantly so. The pattern of findings suggests parallels with the broader society. PMID- 10804700 TI - Responding to aggressive behavior: impact of training on staff members' knowledge and confidence. AB - The impact of a training program designed to help staff members prevent and effectively manage aggressive behavior was assessed with 109 staff members who worked in services for persons with developmental disabilities. Instruction in distraction and defusion strategies was provided as was nonaggressive physical intervention. We employed both a between- and within-participant design. Results indicated that the training significantly improved participants' knowledge of reactive behavior management and their confidence to work with aggressive individuals. PMID- 10804701 TI - A brief overview of Social Role Valorization. AB - Social roles dominate people's lives, and people largely perceive themselves and each other in terms of their roles. The value people attribute to various social roles tends to decisively shape their behavior toward persons whom they see in valued or devalued roles. Those in valued roles tend to be treated well and those in devalued roles, ill. The most current and recently revised version of the Social Role Valorization (SRV) schema is presented in condensed form, showing how social role theory can be recruited for designing very powerful practical measures to pursue valued roles for mentally retarded and other persons or classes at risk of social or even societal devaluation, to upgrade the perceived value of the roles such persons already occupy, and/or to extricate such persons from devalued roles. PMID- 10804702 TI - Analysis of the "Learning in Regular Classrooms" movement in China. AB - The Learning in Regular Classrooms experiment has evolved in response to China's efforts to educate its large population of students with disabilities who, until the mid-1980s, were denied a free education. In the Learning in Regular Classrooms, students with disabilities (primarily sensory impairments or mild mental retardation) are educated in neighborhood schools in mainstream classrooms. Despite difficulties associated with developing effective inclusive programming, this approach has contributed to a major increase in the enrollment of students with disabilities and increased involvement of schools, teachers, and parents in China's newly developing special education system. Here we describe the development of the Learning in Regular Classroom approach and the challenges associated with educating students with disabilities in China. PMID- 10804703 TI - Effects of using a photographic cueing package during routine school transitions with a child who has autism. AB - Making successful transitions from one activity to another is difficult for many children, particularly those who have cognitive, language, or behavioral disabilities. Appropriately terminating one activity and initiating another in a timely fashion is a skill important for young children to learn prior to entering kindergarten. The efficacy of teaching a young child labeled as having autism to make successful transitions in daily routines in three different school settings through the use of photographic cue package was examined. A multiple baseline across-settings design was used to evaluate the efficacy of the intervention. Implications for the use of photographic cue packages in teaching a variety of activities to young children are discussed. PMID- 10804704 TI - Successful friendships of Hispanic children and youth with disabilities: an exploratory study. AB - Interviews were conducted with Hispanic children who had a disability and a friend with whom they had a successful friendship, as well as with parents and teachers. We focused the interviews on the children's and adults' descriptions of the friendship, what each of the children gave to and received from the friendship, the evolution of the friendship in terms of intensity, and the influence of Hispanic cultural values. A friendship support conceptual framework is presented to organize the data related to three relationship domains (companionship, instrumental support, and emotional support) and three friendship depth levels (acquaintance, casual, and intimate). We focused our key recommendations on future directions for research and friendship facilitation. PMID- 10804705 TI - Attitudes of high school students toward individuals with mental retardation. AB - The attitudes of 144 high school students toward individuals who have mental retardation were examined using the Mental Retardation Attitude Inventory-Revised (Antonak & Harth, 1994). The inventory is divided into four subscales consisting of statements regarding integration-segregation, private rights, social distance, and subtle derogatory beliefs. Each subscale was assessed along with gender, frequency of contact with individuals who have mental retardation, and prior knowledge about mental retardation. Results indicate that gender and frequency of contact does influence attitudes. These results are significant because these high school students either are or will soon become adults and impact the community with their attitudes. PMID- 10804706 TI - Contaminated and unusual food combinations: what do people with Prader-Willi syndrome choose? AB - Although hyperphagia is a salient feature of Prader-Willi syndrome, researchers have yet to move beyond food preference and taste studies to examine more troublesome food-seeking behaviors in this population (e.g., eating food from the trash, eating inappropriate or unpalatable food combinations). Visually based tasks were used to examine willingness to eat contaminated food and various odd food combinations in 50 adolescents and adults with Prader-Willi syndrome, 42 IQ matched participants with mental retardation, and 50 control participants without mental retardation. Although participants with Prader-Willi syndrome showed understandings similar to those of control participants about the purpose and fate of food, they endorsed eating contaminated food as well as highly unusual edible and inedible food combinations. Findings suggest novel adjuncts to traditional dietary approaches in the Prader-Willi syndrome population. Implications are also discussed for future research on food ideation in this population. PMID- 10804707 TI - Execution of people with mental retardation: a violation of National and Customary International Law. PMID- 10804708 TI - Why individualized habilitative plans may fail: when challenging behaviors are symptoms of a psychiatric disorder. PMID- 10804709 TI - Growth in state commitments for community services: significance of the Medicaid Home and Community-Based Services Waiver. PMID- 10804710 TI - [Mammography screening. Senselessly squandered money?]. PMID- 10804711 TI - [The physician as a risk factor. 300 women lost their healthy breasts]. PMID- 10804712 TI - [Colonoscopy as intestinal screening: patients will not accept the procedure]. PMID- 10804713 TI - [Sex addiction diagnosis. When libido becomes a burden. Interview by Dr. Beate Schumacher]. PMID- 10804714 TI - [Psychiatric emergencies. Don't depend on your well-trained body!]. PMID- 10804715 TI - [Patient suffers from insomnia. Proscribing bed rest]. AB - Roughly every fifth patient attending the doctor's office complains of insomnia. The first thing that needs to be done is to determine whether a particular patient has primary insomnia of pathological significance, or a temporary disturbance of his/her sense of well-being, or whether the sleep disorder is a secondary consequence of some other somatic or psychiatric condition. For the treatment of insomnia in the doctor's office, a number of basic rules derived from behavioral medicine can be recommended, for example systematic self observation and record-keeping in a sleep diary, reassurance as to the harmlessness of a temporary sleep problem, practicing relaxation to reduce the general level of activity, reduction of the time spent in bed, and sleep deprivation treatment. For all of these forms of treatment, a good doctor-patient relationship is mandatory, in particular when--as is to be expected--setbacks occur, which should not be dramatized. A diagnosis-treatment diagram is presented, in which proposals for medication should also be integrated. PMID- 10804716 TI - [Disturbed sleep caused by sleep apnea. To the sleep laboratory for diagnosis?]. AB - Since it was shown that numerous neurological, psychiatric and internal illnesses have characteristics manifested during, or influenced by sleep, somnology has been playing a clinically more and more important role. Among the 88 diagnoses listed by ICD, not only insomnia, but also sleep-related respiratory disorders, in particular the obstructive sleep apnea syndrome, are of special importance. Sleep apnea is associated with coronary heart disease, myocardial insufficiency and other pathological conditions. Already in the doctor's office, a carefully taken history (nocturnal apnea alternating with irregular snoring, and diurnal sleepiness) can arouse an appropriate suspicion. This can be confirmed by an ambulatory polygraphic exploration. The definitive diagnosis is then established with the aid of polysomnography in the sleep lab where specific treatment is also initiated. PMID- 10804717 TI - [Chronic sinusitis--operation per endoscope. Hope for freedom from symptoms]. PMID- 10804718 TI - [The higher, the earlier. Hypertension causes dementia]. PMID- 10804719 TI - [Is the alcoholic really sober, does the applicant drink, can the pilot still fly? CDT test often fails in reliability!]. PMID- 10804720 TI - [Therapy of gout. Management of an acute attack--secondary prevention]. PMID- 10804721 TI - [Facial exanthema with joint symptoms. Systemic lupus erythematosus]. PMID- 10804722 TI - [Physicians and insurance against consumer organizations. Squandered money for senseless bureaucracy]. PMID- 10804723 TI - [Azathioprine also in chronic active ulcerative colitis. Brake pedal for the immune system]. PMID- 10804724 TI - Partial left ventriculectomy in children and adolescents. AB - Theoretical aspects, pediatric indications and initial results of a new operation, partial left ventriculectomy, are presented. PMID- 10804725 TI - Adult survivors of children's cancer and their offspring. AB - BACKGROUND: Although it is anticipated in Japan that the number of long-term survivors of children's cancer will rapidly increase and that they will have children, reports of studies concerning the offspring of such survivors have come mainly from western countries. For this reason, it seems that the results of this study will be important. METHODS: Ninety-seven survivors of children's cancer, who were diagnosed between 1962 and 1989 and are now older than 20 years old, and their offspring were studied. RESULTS: Of 97 survivors, 30 have married and 23 survivors or their spouses had been pregnant (33 total pregnancies). Twenty-five babies were born and seven pregnancies ended in spontaneous abortion. The abortion rate was not different from that of the Japanese general population. The birthweights of babies born to survivors tended to be lower than those of control subjects. The proportion of babies with birthweights under 2500 g was significantly greater for babies born to survivors than to the control subjects. None of the 25 babies born had congenital anomalies. Subsequent development of these children has been uneventful with no malignancies after a median follow-up period of 4 years 10 months (range: 10 months to 16 years, 3 months). CONCLUSIONS: The reproductive activity of children's cancer survivors and the health status of their offspring seem to be satisfactory. However, the sample size of the present study is too small to draw any definite conclusions. Because it is anticipated that the number of long-term survivors of children's cancer will rapidly increase in Japan, it is important to establish an effective system of following up these survivors and their offspring in order to provide them with appropriate suggestions for a better quality of life. PMID- 10804726 TI - Regulation of CD31 expression and interleukin-4 production by human cord blood CD4+ T cells with interleukin-4 and interleukin-7. AB - BACKGROUND: T helper 2 cell type cytokines, such as interleukin (IL)-4 and IL-5, play pivotal roles in the development of allergic diseases. However, the mechanism by which naive CD4+ T cells acquire the ability to produce these cytokines remains unclear. Recently, it was reported that IL-7 induces the ability to produce IL-4 as well as interferon (IFN)-gamma and IL-5 in naive CD4+ T cells without TCR stimulation. To further analyze the mechanism of acquiring IL 4-producing ability by naive CD4+ T cells, the effects of IL-7 on human cord blood CD4+ T cells were compared with those of IL-4, which induced the ability to produce IFN-gamma but not IL-4. RESULTS: Interleukin-7 preserved the population of CD4+CD31- T cells in cord blood and induced their IL-4-producing ability without T cell receptor (TCR) stimulation, while IL-4 induced CD31 on CD31- T cells and could not induce their IL-4-producing ability. Both the CD31-inducing effect and the inhibitory priming effect for IL-4-production by IL-4 were also observed after cord blood CD4+ T cells had been primed with IL-7 and acquired the IL-4-producing ability. CONCLUSIONS: Interleukin-7 induced the IL-4-producing ability in naive CD4+CD31- T cells without TCR stimulation, suggesting that the signal transduction via CD31 may have an inhibitory effect on the acquisition of the IL-4-producing ability by cord blood CD4+ T cells in the absence of TCR stimulation. PMID- 10804727 TI - Echocardiographic evaluation of the ductal morphology in patients with refractoriness to lipo-prostaglandin E1 therapy. AB - BACKGROUND: Lipo-prostaglandin (PG)E1 is effective at lower doses and has fewer side effects than PGE1-cyclodextrin (CD). Previous studies, however, have suggested that some patients show refractoriness to lipo-PGE1 in the course of treatment. The present paper examines: (i) whether such cases can be predicted by examining the ductal morphology before and 24 h after the start of lipo-PGE1 infusion; and (ii) whether PGE1-CD dilates the ductus arteriosus in patients with refractoriness to lipo-PGE1. METHODS: The ductal morphology was evaluated with two echo indices, such as minimal and minimal plus maximal intraluminal diameters of the ductus. Two-dimensional echocardiography was performed in 24 patients with ductus-dependent congenital heart disease. The two echo indices were measured before and 24 h after lipo-PGE1 infusion and also at least twice per week until surgery. RESULTS: In 19 of 24 patients, ductal patency was maintained until surgical treatment (group A). The remaining five patients (21%) showed ductal closure during the course of the lipo-PGE1 therapy (group B). There were no significant differences between the two groups, in either the maximal or minimal diameters, which were examined before and 24 h after treatment. In the five patients of group B, lipo-PGE1 was replaced with a relatively high dosage of PGE1 CD (50-100 ng/kg per min), resulting in good ductal patency until surgery. CONCLUSIONS: Patients with refractoriness to lipo-PGE1 therapy could not be predicted from initial intraluminal diameters of the ductus using echocardiography. Therefore, serial echocardiographic examinations are important to detect early findings of ductal closure. In addition, PGE1-CD is still useful as back-up therapy in such patients. PMID- 10804728 TI - Anti-interleukin-8 auto-antibodies in cerebrospinal fluid of children with purulent meningitis. AB - BACKGROUND: Cerebrospinal fluid (CSF) of patients with purulent meningitis contains a high concentration of interleukin (IL)-8. Recently, the presence of anti-IL-8 auto-antibodies was noted in blood and alveolar fluid. Therefore, measurement of the concentration of anti-IL-8 auto-antibodies was attempted in CSF of children with and without meningitis. METHODS AND RESULTS: We measured the concentration of anti-IL-8 auto-antibodies in CSF of children with purulent or aseptic meningitis and those without meningitis. The CSF obtained on admission showed a significantly higher concentration of anti-IL-8 IgG and IgM auto antibodies in children with purulent meningitis, compared with those with aseptic meningitis or without meningitis. Among the three groups of children, the concentration of IL-8 was also significantly higher in CSF of children with purulent meningitis. CONCLUSION: Because the anti-IL-8 IgG auto-antibody binds to IL-8 and inhibits IL-8 interaction with specific receptors on neutrophils, the presence of anti-IL-8 auto-antibodies seems to provide a mechanism that limits the bioavailability of free IL-8 in CSF. PMID- 10804729 TI - Influence of emission from rice straw burning on bronchial asthma in children. AB - BACKGROUND: Emission from rice straw burning (ERSB) is observed everywhere after harvest of rice in Niigata Prefecture every year. Pediatricians and many guardians in this district have had the impression that ERSB may induce asthma attack. Recent studies have suggested that particulate air pollution plays a role in the exacerbation of asthma. The authors investigated relationship of ERSB to asthma attack in children. METHODS: A questionnaire on rice straw burning (RSB) was circulated to guardians and pediatric institutions. Change in the monthly number of children with asthma attack (CAA) for 5 years from January 1994 to December 1998 was investigated. In addition, change in the number of CAA from the meteorologic conditions and RSB was investigated from the fourth week of August to the third week of September in 1996, 1997 and 1998. Challenge test exposure to ERSB was tried on a volunteer adult with chronic asthma. The situation of air pollution was examined by measuring suspended particulate matter (PM10). The relationship between PM10 and the number of CAA was studied. RESULTS: A majority of the guardians had the impression that ERSB induces asthma attack. Pediatricians replied similarly to the questionnaire. The number of CAA visiting our emergency room and admitted to our ward increased in the season of RSB. The PM10 had a significant correlation with the number of CAA. It was suggested that the increase in CAA may be not due to the meteorologic conditions, but to the influence of ERSB. CONCLUSION: The ERSB has made an issue of air pollution. Furthermore, the possibility that ERSB induces or exacerbates asthma attack has become clear in the present study. Therefore, it is recommended that RSB should be abolished for the health of inhabitants, especially children with asthma. PMID- 10804730 TI - Effects of growth hormone replacement therapy on lipids, lipoproteins and apolipoproteins: is the increased apolipoprotein A-1/B ratio the only benefit? AB - BACKGROUND: The aim of the present study was to investigate the controversial role of growth hormone (GH) therapy in lipid regulation. METHODS: We studied serum levels of cholesterol and subgroups, triglyceride and apolipoprotein A-1 and B in 41 GH-deficient children (with subgroups of untreated and short- and long-term treated subjects) and 20 healthy controls. RESULTS: Cholesterol and low density lipoprotein cholesterol levels (in mmol/L) were found to be 4.92 +/- 1.34 and 3.02 +/- 1.58 in untreated, 4.15 +/- 0.72 and 2.46 +/- 0.65 in short-term (3 month) treated, 4.93 +/- 1.39 and 3.15 +/- 1.38 in long-term (> 1 year) treated and 4.11 +/- 0.5 and 2.0 +/- 0.74 in control subjects, respectively. The apolipoprotein A-1:B ratio was 1.98 +/- 0.5 in long-term treated and 1.6 +/- 0.6 in control subjects. CONCLUSIONS: The improvement of lipid composition with short term GH therapy is temporary, but the increase in apo A-1:B is not and seems to be the particular benefit of this therapy. PMID- 10804731 TI - Preperitoneal fat thickness in childhood obesity: association with serum insulin concentration. AB - BACKGROUND: The clinical significance of visceral fat accumulation in childhood obesity has remained controversial. METHODS: In 35 subjects (18 boys and 17 girls) aged 9-15 years, visceral and subcutaneous fat were estimated by measuring the maximum thickness of preperitoneal fat (Pmax) and the minimum thickness of subcutaneous fat (Smin) in the upper median abdomen, respectively, by ultrasonography. The associations with fasting serum insulin levels converted to logarithm (log(IRI)) were studied for six obesity-related indices, including the obesity index, body fat percentage, waist-to-hip circumference ratio (WHR), Pmax, Smin and the ratio of Pmax to Smin (abdominal wall fat index, AFI). RESULTS: In girls, the log(IRI) was positively correlated with the Pmax, obesity index, body fat percentage and Smin. In contrast, the log(IRI) was positively correlated only with the Pmax and Smin in boys. There were no significant correlations between the log(IRI) and the WHR or AFI in either sex. The Pmax revealed the highest correlation with the log(IRI) in both boys (r = 0.703, P < 0.001) and girls (r = 0.921, P < 0.0001). CONCLUSION: These results indicate the important contribution of visceral fat accumulation to the development of hyperinsulinemia, regardless of sex. Accordingly, it is suggested that evaluating Pmax by ultrasonography may be the most sensitive and reliable method of predicting insulin resistance associated metabolic derangements in children, compared with other conventional indices of obesity. PMID- 10804732 TI - Sporadic case of hemoglobin Bushwick detected by chance in aplastic crisis. PMID- 10804733 TI - Case of transient erythroblastopenia of childhood with transient hypogammaglobulinemia. PMID- 10804734 TI - Hemophagocytic lymphohistiocytosis with preceding neurologic signs and neutrophilia. PMID- 10804735 TI - Myeloperoxidase antineutrophil cytoplasmic antibody positive vasculitis during propylthiouracil treatment: successful management with oral corticosteroids. PMID- 10804736 TI - Sepsis and meningitis due to penicillin-resistant viridans streptococci in neutropenic children. PMID- 10804737 TI - Infantile exanthematous disease with elevated anti-toxic shock syndrome toxin-1 antibody: related to toxic shock syndrome toxin-1? PMID- 10804738 TI - Pediatric case of hernia reduction en masse. PMID- 10804739 TI - Three patients with different phenotypes in a family with chromosome 22q11.2 deletions. PMID- 10804740 TI - Influenza surveillance system of Japan and acute encephalitis and encephalopathy in the influenza season. AB - AIM: To understand the situation of influenza epidemics and neurologic complications during the influenza season in Japan. METHODS: The influenza surveillance system in Japan was reviewed and the data on influenza/influenza like illness and neurologic complications, such as acute encephalitis/encephalopathy, were evaluated. RESULTS: From the data of the National Epidemiological Surveillance of Infectious Diseases (NESID), an increase in clinically diagnosed encephalitis/encephalopathy was clearly noted recently. Most cases reported were acute encephalitis/encephalitis. The number of cases of Reye's syndrome reported was stable and low in number, since the time of the recommendation of not using aspirin for pediatric febrile disease, particularly for influenza-like illness, in Japan. These acute encephalitis/encephalopathy cases were not correlated clearly with clinically diagnosed influenza patients before the 1997/98 influenza season from the surveillance data, although several case reports have shown a relationship. CONCLUSIONS: It is clarified from the NESID that neurologic complications such as acute encephalitis/encephalopathy, apart from Reye's syndrome, have occurred associated with influenza virus infection, especially among young children. Further investigation should be necessary to find the pathogenesis of this serious complication of influenza virus infection and whether this is unique in Japan, due to unclarified factor/factors, or whether it has not been recognized in other countries. PMID- 10804741 TI - Epidemiology of influenza-associated encephalitis-encephalopathy in Hokkaido, the northernmost island of Japan. AB - BACKGROUND: It is well known that acute onset brain dysfunction, which usually is diagnosed as encephalitis or encephalopathy, occurs in association with influenza. However, this may have been underestimated as a rather infrequent event. Sixty-four infants and children developed encephalitis-encephalopathy during the five recent influenza seasons in Hokkaido, the northernmost island of Japan. METHODS: Inquiries were sent at the end of each season, from October 1994 to March 1999, to 94 hospitals and institutes in Hokkaido which accept pediatric age patients, asking if there were any admitted cases of encephalitis or encephalopathy. RESULTS: The patients were 42 boys and 22 girls and 47 (73.4%) were 4 years of age or younger. None of them had received an influenza vaccine nor had an oral administration of aspirin. Most of the patients became comatose with or without convulsions within a few days of the onset of fever. Twenty-eight (43.8%) patients died and 13 (20.3%) had neurological sequelae. Patients with clotting disorders, elevations of serum creatine kinase and/or aspartate aminotransferase and alanine aminotransferase, and brain CT abnormalities had a poor prognosis compared with patients without. Among these affected patients, the influenza genome (H3) was detected by polymerase chain reaction in nine cerebrospinal fluid samples, influenza virus A (H3N2) was isolated in 18 nasopharyngeal swab samples and a four-fold or greater rise in serum hemagglutinin inhibition antibody titer against H3N2 was observed in seven patients. CONCLUSIONS: It appears urgent to promote vaccination against influenza in young children to prevent these devastating disease conditions. PMID- 10804742 TI - Hypothetical pathophysiology of acute encephalopathy and encephalitis related to influenza virus infection and hypothermia therapy. AB - BACKGROUND: To establish a treatment strategy for acute encephalopathy and encephalitis associated with influenza virus infection, the pathophysiology of the disease was investigated through manifestations and laboratory findings of patients. PATIENTS AND METHODS: A child with central nervous system (CNS) complications during the course of influenza virus infection was analyzed in view of immunologic abnormalities. In addition, four children with acute encephalopathy and encephalitis were enrolled in the hypothermia treatment for the purpose of stabilizing the cytokine storm in the CNS. RESULTS: The CNS symptoms preceded the systemic progression to the failure of multiple organs (MOF) and disseminated intravascular coagulopathy (DIC). The mild hypothermia suppressed the brain edema on computed tomography (CT) scanning and protected the brain from the subsequent irreversible neural cell damage. CONCLUSION: The replicated viruses at the nasopharyngeal epithelium may disrupt the olfactory mucosa and gain access to the brain via the olfactory nerve system. The direct virus-glial cell interaction or viral stimulation of the glial cells induces the production and accumulation of the pro-inflammatory cytokines, especially tumor necrosis factor (TNF)-alpha, in the CNS. The cytokine storm results in neural cell damage as well as the apoptosis of astrocytes, due to the TNF-alpha-induced mitochondrial respiratory failure. The disruption of the blood-brain barrier progresses to the systemic cytokine storm, resulting in DIC and MOF. Mild hypothermia appears promising in stabilizing the immune activation and the brain edema to protect the brain from ongoing functional, apoptotic neural and glial damage and the systemic expansion of the cytokine storm. PMID- 10804743 TI - Influenza virus-induced encephalopathy: clinicopathologic study of an autopsied case. AB - BACKGROUND: Rapid progressive encephalopathy with a high fever, consciousness loss and recurrent convulsions has been occasionally reported in children during influenza pandemics in Japan since 1995. We examined a 2-year old girl with hemorrhagic shock and encephalopathy syndrome associated with acute influenza A virus infection (A/Nagasaki/76/98; H3N2), to answer several questions for which no histologic or virologic data exist. METHODS: A clinicopathologic study using immunohistochemical staining and viral genome detection by reverse transcriptase polymerase chain reaction (RT-PCR) was performed with this autopsied case. RESULTS: The virus antigen was positive in CD8+ T lymphocytes from the lung and spleen. The virus infected a very limited part of the brain, especially Purkinje cells in the cerebellum and many neurons in the pons, without inducing an overt immunologic reaction from the host. The RT-PCR used for detecting the hemagglutinin gene demonstrated positive bands in all frozen tissues and cerebrospinal fluid taken at autopsy and not in samples obtained on admission. CONCLUSIONS: The pathologic change induced by the direct viral invasion cannot be responsible for all of the symptoms, especially for the rapid and severe clinical course of the disease within 24-48 h after the initial respiratory symptoms. Together with the rapid production of several inflammatory cytokines, the breakdown of the blood-brain barrier may induce severe brain edema and can be a major pathologic change for the disease. Any therapeutic strategy to control this multistep progression of the disease could be effective. PMID- 10804744 TI - Influenza-associated encephalopathy in Japan: pathogenesis and treatment. AB - It is estimated that more than 100 children die of influenza-associated encephalopathy (influenza encephalopathy) every year in Japan. Influenza encephalopathy is distinct from Reye's syndrome. Specifically, 20% of influenza encephalopathy patients exhibit bilateral thalamic necrosis on neuroimaging, a lesion referred to as acute necrotizing encephalopathy (ANE). Influenza encephalopathy may develop by the same pathogenetic mechanisms as ANE, possibly via vasoactive substances or a process leading to vasoconstriction in the central nervous system (CNS). A novel substitution at the receptor-binding site (Tyr 137 to Phe) was reported to be found exclusively in influenza type A (H3N2) viruses isolated from patients with influenza encephalopathy, suggesting that encephalopathy may be caused by a variant influenza type A (H3N2) virus. Recently, it has been reported that cytokines may mediate the disease and that a high plasma concentration of interleukin-6 could be an indicator of progression to encephalopathy. Although it is unknown whether influenza virus invades the CNS, amantadine therapy for influenza encephalopathy has been tried in Japan, in patients in whom influenza type A infection has been demonstrated by rapid antigen detection tests. PMID- 10804745 TI - Vaccines and therapeutics against influenza virus infections. AB - BACKGROUND: Vaccination prior to epidemic season is the best measure to control influenza virus infection; however, there are several issues to be considered regarding influenza vaccines in Japan. In the present review, current issues regarding influenza vaccine in Japan are described, as well as the future prospects of vaccine development. As well as vaccination, anti-influenza agents such as amantadine are now available in Japan. Furthermore, neuraminidase inhibitors are expected to appear in the market in near future. These anti influenza agents represent new options for influenza control. CONCLUSIONS: Vaccination and antiviral agents are a major armamentarium against influenza infections. Thus, exploratory studies on novel forms of vaccine and anti influenza drugs should help to prepare against pandemics that must emerge in near future. PMID- 10804746 TI - Postural change and pulmonary ventilation-perfusion distribution after Fontan operation. PMID- 10804747 TI - [Primary HIV drug resistance]. PMID- 10804748 TI - [The treatment of Parkinson disease. An update and the outlook]. PMID- 10804749 TI - [The epidemiology and modes of transmission of Helicobacter pylori infection]. PMID- 10804750 TI - [Can Helicobacter pylori infection be a risk factor for the severity of rheumatoid arthritis?]. AB - AIM: In this study we assessed the effect of Helicobacter pylori eradication in patients with established rheumatoid arthritis (RA) in order to show a possible pathogenetic role of the infection in this disease. METHODS: We recruited 31 patients with variable RA activity. Of them, 15 were Hp-positive (12 F and 3 M, mean age 55 +/- 10.6 years) and 16 Hp-negative (13 F and 3 M, mean age 54.2 +/- 9.1 years) on the basis of the concomitant positive or negative findings of both CLO-test and histology performed on both antral and corpus biopsies. The infected group was treated and the bacterium was considered eradicated when both tests were negative a month after therapy. We have evaluated the disease activity at baseline and during a total follow-up period of 16 months with check-points every 4-months and compared clinical and laboratory findings between the Hp-negative and the eradicated groups. Both groups were being treated with NSAIDs and prednisolone (< or = 7.5 mg/die) or its equivalents. RESULTS: Hp-eradicated RA patients showed a progressive improvement overtime (p = 0.0009) of all clinical indices compared with baseline, while Hp-negative RA patients did not. At the 16 month checkpoint, the eradicated RA patients differed significantly (p < 0.006) from patients without Hp infection by all indices, except grip strength of the hands. Also all laboratory data improved significantly from baseline to the 16 month checkpoint (p < 0.03) within the Hp-eradicated group and between the two groups of eradicated and Hp-negative RA patients (p < 0.0007) except for Hb, aCL IgM and gamma-globulins. CONCLUSIONS: Our data suggest that Hp infection is implicated in the pathogenesis of RA. On the basis of our data, the eradication of Hp should be recommended in infected RA patients. PMID- 10804751 TI - [Gastric MALT lymphoma and Helicobacter pylori]. AB - Aim of this paper is to evaluate the effects of eradicating Helicobacter pylori (Hp) on the course of low-grade gastric mucosa-associated lymphoid tissue (MALT) lymphoma. A total of 13 consecutive patients, 7 males and 6 females (median age 54 years) Hp-positive were included, all affected by low-grade B-cell gastric MALT lymphoma, stage I using established criteria; histologic assessment by gastric biopsies was performed. Each patient received anti-Hp therapy with standard protocol for 7 days and underwent endoscopy and histology every 3-6 months. Complete eradication was obtained in all patients (100%); on histology 9 patients showed complete regression, but in one of these lymphoma relapsing after 12 months was observed. Our findings confirm recent cooperative reports of Hp eradication rate and regression of gastric tumour (about 70%). In conclusion, the indolent clinical course of the disease justifies a first-line conservative approach with Hp eradication therapy. PMID- 10804752 TI - [The physiopathological aspects of chronic gastritis, Helicobacter pylori and precancerous gastric changes]. AB - Chronic gastritis represents the paradigmatic histological condition by means of which the various clinical displays develop. Among the several etiologic factors, Helicobacter pylori (Hp) certainly represents a determining element for the promotion of such condition. Several morpho-functional evaluations (parietal cells, peptic cells, acid secretion, peptic secretion, gastrinemia) point out how Hp has on the mucosa a mainly direct action, without any substantial modifications of the gastric milieu. Certainly, the bacteria help the bodyfundic atrophic gastritis, but seldom this condition contributes to the modification of the gastric pH in such a relevant way so as to prime the fall of events which lead to the preneoplastic/neoplastic modifications. Metaplastic, dysplastic and neoplastic clones may arise, in the course of Hp infection, even in conditions of normochloridria. Such bacterium behaves as a cofactor of various other damaging or predisposing agents, contributing to modulate the risk by acting on host genetic susceptibility. PMID- 10804753 TI - [Lansoprazole: an analysis of the clinical trials in the 3 years of 1997-1999]. AB - Aim of this overview was to evaluate the main clinical trials with lansoprazole published from 1997 to 1999 in English-language journals, regarding gastroesophageal reflux disease, peptic ulcer, NSAID-induced ulcer, and ZES. Results of clinical trials for therapy and prevention of lesions/symptoms have been evaluated separately. In direct comparisons, lansoprazole alone (not combined with antibiotics) proves to be equieffective to other PPI and more effective than H2-RA in both therapy and prevention of GERD, peptic ulcer (a part from anti-Hp regimens) and NSAID-induced ulcer. Among Hp-eradicating regimens in patients with peptic ulcer or functional dyspepsia, lansoprazole-based triple therapy is equal in efficacy to other PPI-based or RBC-based triple therapies and, in any case, significantly better than dual therapies. The in vitro anti-Hp activity of lansoprazole, more marked than with other PPI, does not seem to effort clinical advantages. Safety of lansoprazole is largely satisfactory and no different from other PPI and H2-RA. PMID- 10804754 TI - [Influenza in the 21st century: preparing ourselves for a new pandemic]. PMID- 10804755 TI - [Advanced rectal cancer. Its evolution and the current situation of adjuvant treatment]. PMID- 10804756 TI - [Rectal cancer. The effect of the clinical stage and the degree of lymphatic invasion on local recurrence and survival. An analysis of 168 patients]. AB - INTRODUCTION: Surgery is the most important therapeutic means for treatment of rectal carcinoma. Nevertheless, from stage B2, relapse rates are high and it is therefore necessary to use supplemental treatments such as radiotherapy associated or not with chemotherapy OBJECTIVE: To assess the likelihood of being free of local, local and distant disease and overall and specific survival in function of clinical stage and degree of lymph node involvement among patients diagnosed with colo-rectal adenocarcinoma treated with radical surgery and radiotherapy associated or not with chemotherapy. MATERIALS AND METHODS: Since January 1990 up to December 1997, all patients with rectal adenocarcinoma treated with radical surgery and postoperative radiotherapy, with or without chemotherapy, were prospectively included in a database which was analyzed. RESULTS: The crude actuarial survival at five years was 61.1 +/- 9.2% and specific survival 64.2 +/- 9.2%. As for stages: B (84.1 +/- 10.1%) and C (49.9 +/ 3.3%) (p < 0.001). Likewise, for N0 84.1 +/- 10.1%, for N1 62.2 +/- 13.5% and for N2 13.7 +/- 22.3% (p < 0.001). The likelihood of being in complete remission for the overall patient population was 50.2 +/- 9.2%: B (67.5 +/- 13.5%) and C (37.9 +/- 11.9%) (p < 0.001). Likewise, for N0 67.5 +/- 13.5%, for N1 47.8 +/- 13.5%, and for N2 9.9 +/- 17% (p < 0.001). CONCLUSIONS: Given the poor results obtained in stages C, particularly stage N2 and also that a better local control is obtained with good tolerance when preoperative radiotherapy is administered, we believe that in order to improve the results it is necessary to initiate preoperative radiotherapy. PMID- 10804757 TI - [The risk factors associated with colonization and bacteremia in non-tunnelled central venous catheters]. AB - OBJECTIVE: To identify risk factors for colonization and bacteremia among patients with non-tunnelled central venous catheters. MATERIALS AND METHODS: A prospective study was conducted of a cohort of patients carrying non-tunnelled central venous catheters. Different parameters were obtained and the degree of its association with colonization of the distal portion of the catheter or with bacteremia associated with colonization was estimated. The CDC (centers for Disease Control) diagnostic criteria of colonization and catheter-related bacteremia were used. RESULTS: A total of 118 catheters were eventually analyzed, corresponding to 114 patients, with a catheterization mean time of 14 +/- 8 days (mean +/- SD); out of these 114 patients, 51 were colonized and in 22 the presence of associated bacteremia was confirmed. The parameters associated with a higher risk for catheter colonization included length of colonization, femoral location, number of lumina and a vital prognosis lower than one month. All these factors, with the exception of the increase in the number of lumina, showed an independent association with colonization on the multivariate analysis [catheterization length (in weeks): OR 1.46; 95% CI: 1.0-2.11; femoral location: OR 3.73; 95% CI: 1.16-11.9; vital prognosis lower than one month: OR 12.7; 95% CI: 1.4-112.7]. As for risk for catheter-related bacteremia, the univariate analysis showed an association with catheterization length and a vital prognosis lower than one month; the latter was the only factor that maintained an independent association in the multivariate analysis (OR 5.75; 95% CI: 1.17 28.27). CONCLUSION: The present study documents the relevance of prolonged catheterization as a consistent risk for colonization of non-tunnelled central venous catheters. This risk increases independently in canalization at femoral site and particularly among severely ill patients. The presence of these factors allows the identification of a high risk population for the development of catheter related bacteremia. PMID- 10804759 TI - [Abdominal infection by Pasteurella spp. A report of 3 cases]. AB - Pasteurella spp. is a gram-negative coccobacillary micro-organism which is a common cause of soft-tissue infections caused by animal bite or scratch and more uncommonly at another locations. We report here the clinical and evolutive characteristics of three patients with different varieties of intra-abdominal infection caused by the microorganism as well as the potential exposure to pet animals. PMID- 10804758 TI - [43 cycles of anabolic steroid treatment studied in athletes: the uses and secondary effects]. AB - OBJECTIVE: To know the actual abuse of anabolic steroids by amateur athletes in our environment as well as actions and secondary effects resulting from such abuse. METHODS: Analytical observational study from May 1997 to November 1998. Forty-three therapy courses with anabolic steroids among 39 male athletes were studied. Diet and training were standardized for all participants. A verification was made that the test group started from a basal state. RESULTS: Duration of therapy was 6 weeks and the mean total dose was 2,928 mg. Significant differences were found in the test group regarding basal and post-therapy values for: transaminases (AST: 29.8 vs 45.0 IU/l, p < 0.001. ALT: 32.9 vs 51.4 IU/l, p < 0.01), cholesterol, HDL-cholesterol (31.4 vs 19.7 mg/dl, p < 0.01), LDL cholesterol (145.9 vs 173.5 mg/dl, p < 0.01), LH (2.1 vs 0.2 U/l, p < 0.001), FSH (3.3 vs 0.4 U/l, p < 0.001), free testosterone (14.4 vs 34.0 pg/ml, p < 0.001), 17-beta-estradiol and arm muscular section (98.8 vs 103.7 cm2, p < 0.001). The inclusion of testosterone in therapy introduced a significant difference with respect to the use of synthetic anabolic agents alone, in total testosterone (4.5 vs 0.9 ng/ml, p < 0.001) and 17-beta-oestradiol, but neither with respect to free testosterone nor arm muscular section. An 84.6% of individuals in the problem group stated to complete two therapy courses in a year. CONCLUSIONS: The use of anabolic steroids increases the lean muscular mass. The most relevant secondary effects included: increased transaminase serum levels, change in the lipid profile and suppression of the hypothalamus-pituitary gland-gonad axis. The inclusion of testosterone did not increase the lean muscular mass. PMID- 10804760 TI - [Screening for colorectal cancer]. PMID- 10804761 TI - [New drugs for the treatment of type-2 diabetes]. PMID- 10804762 TI - [The transplantation of solid organs in Spain: the success of a policy]. PMID- 10804763 TI - [A 51-year-old patient with progressive neurological deterioration following a liver transplant]. PMID- 10804764 TI - [A 35-year-old woman with visual perseveration phenomena in the left homolateral hemifield]. PMID- 10804765 TI - [Syncope with hypocalcemia]. PMID- 10804766 TI - [Dorsalgia in a young woman]. PMID- 10804767 TI - [Yellowish papules on the extremities associated with bone lesions]. PMID- 10804768 TI - [An adult male with an old inferior myocardial infarct, normal global ventricular function and repetitive episodes of sustained wide-QRS tachycardia]. PMID- 10804769 TI - [Endocarditis due to Gemella morbillorum in a parenteral drug-abuse addict]. PMID- 10804770 TI - [Bronchiolitis obliterans organizing pneumonia associated with cutaneous vasculitis]. PMID- 10804771 TI - [Stevens-Johnson syndrome due to nevirapine]. PMID- 10804772 TI - [Diabetes mellitus after the resection of an insulinoma]. PMID- 10804773 TI - [Recurrent subdural hemorrhage as the form of presentation of a type-IV Ehlers Danlos syndrome]. PMID- 10804774 TI - [Acute lithiasic pancreatitis in patients treated with somatostatin analogs]. PMID- 10804775 TI - Primary angioplasty for acute myocardial infarction complicated by cardiogenic shock on admission. AB - AIM: To evaluate primary angioplasty results for the treatment of acute myocardial infarction complicated by cardiogenic shock on admission. POPULATION AND METHODS: Retrospective analysis of 11 consecutive patients with acute myocardial infarction complicated by cardiogenic shock (defined as systolic blood pressure below 80 mmHg and signs of hypoperfusion, despite volume expanders and/or vasopressors infusion) treated with primary angioplasty. Clinical characteristics, angiographic data, hospital outcome and follow-up were analysed. RESULTS: There were ten males (90.9%) with a mean age of 66 years. Eight patients had anterior wall myocardial infarction and three patients had inferior wall myocardial infarction, two of which with extension to the right ventricle. The mean time between symptom onset and angioplasty was 3.5 hours. Three patients had left main coronary artery occlusion; three patients had single vessel disease and five patients had multivessel disease. The angiographic success rate (open infarct-related artery and TIMI III flow) was 90.9% (ten patients). Stent implantation was performed in five patients. Abciximab was given in five patients. In-hospital mortality rate was 36.4% (four patients). The surviving patients had a mean ejection fraction of 43.1% on discharge. In a mean follow-up of 16.2 months, one patient had coronary artery bypass graft and one had stroke. CONCLUSION: Based on published data, our experience with this short series of cases shows that primary angiography should be regarded as a positive option for the treatment of acute myocardial infarction complicated by cardiogenic shock. PMID- 10804776 TI - Primary angioplasty in cardiogenic shock: for every patient? PMID- 10804777 TI - Tissue Doppler imaging in different locations of the mitral annulus: all different or all the same? AB - BACKGROUND: The assessment of the mitral annulus motion with tissue Doppler imaging is claimed to be an accurate method to quantify global left ventricular systolic and diastolic function. However, it is not yet perfectly defined which site of the annulus must be selected. Familial amyloidotic polyneuropathy of the Portuguese type (FAP) is an hereditary systemic disease in which diastolic dysfunction may occur. AIM: 1--To determine if in FAP patients the mitral annulus motion is independent of the selected site. 2--To compare pulsed wave Doppler parameters with tissue Doppler parameters in the different annular sites. METHODS: Of 24 FAP patients studied, 14 were included. In each patient we performed conventional transmitral pulsed wave Doppler and tissue Doppler in the 4 sites of the mitral annulus and measured the velocities of the rapid filling wave e, of the atrial contraction wave a and calculated e/a ratio. RESULTS: According to the transmitral inflow profile, patients were divided in 2 groups: Group I--normal global diastolic function and Group II--abnormal relaxation. Group I--33% of these patients showed e/a > 1 in the four sites and 67% showed e/a > 1 in at least 1, but not in all the sites. The rate of normal sites per patient was 3.1. Group II--25% of these patients showed e/a < 1 in the 4 sites of the annulus and 75% had e/a < 1 in at least 1, but not in all the sites analysed. The rate of abnormal sites/patient was 3.1. in this group. When conventional and tissue Doppler data were compared (bland and altman) the septal portion of the annulus was the one with the best correlation. CONCLUSIONS: 1--The assessment of the mitral annulus motion with tissue Doppler imaging is dependent on the site selected for study. 2--The septal site was the one that showed the highest correlation and concordance between pulsed wave Doppler and tissue Doppler. 3- The relative number of normal versus abnormal sites was determinant of the transmitral pattern. 4--Tissue Doppler imaging identified: a) among patients until now classified as normal diastolic function, a subgroup of patients with abnormal function in some sites of the annulus and b) among patients with abnormal relaxation, a subgroup with normal diastolic function in some sites of the annulus. PMID- 10804778 TI - Influence of salt intake on the daytime-nighttime blood pressure variation in normotensive and hypertensive black subjects. AB - OBJECTIVE: To evaluate the influence of different salt-intake regimens on the circadian rhythm of blood pressure (daytime-night-time relationship) in normotensive and hypertensive black subjects with different patterns of salt sensitivity. METHODS: Randomized, cross-over study. Twenty normotensive (NT) and 27 hypertensive (HT) black subjects were kept on a low-sodium diet (30 mmol sodium/d, LS) and on a high-sodium diet (300 mmol sodium/d, HS) for 1 week each. On the last day of each regimen, 24 hour ambulatory blood pressure monitoring was performed. RESULTS: Eight normotensives were classified as salt-sensitive (SS), all with haptoglobin phenotypes (FeHap) 1,1 or 1,2, and 12 as salt resistant (SR), 5 with FeHap 2,2. Seventeen hypertensives were classified as SS, all with FeHap 1,1 or 1,2, and 11 as SR, 2 with FeHap 2,2. Salt sensitivity criterium was: difference > 5 mmHg of 24 h mean blood pressure from low sodium to high sodium. The pattern of daytime-nighttime blood pressure relationship between LS and HS was only modified (respectively from dipper to non-dipper) in HT-SS, but not in NT-SS, NT-SR and HT-SR. The percentual drop in nighttime mean blood pressure was about 10% in HT-SR and in NT-SR either under LS or HS. In NT-SS, the percentual night-time drop in mean blood pressure was lower than that of NT-RS (i.e. about 7 8%), but it was not different on LS and on HS. In contrast, in HT-SS, the percentual nighttime drop in mean blood pressure on HS (6%) was significantly lower than that on LS (10%, p < 0.01). In the 27 HT, but not in the NT, changes in the nocturnal drop in mean blood pressure induced by salt restriction correlated positively with the degree of salt sensitivity (r = 0.45, p < 0.05). CONCLUSIONS: In black subjects, the pattern of nighttime-daytime blood pressure relationship appears to be modified from LS to HS diets (or vice-versa) only in SS hypertensive subjects, but neither in NT-SS nor in NT-SS and HT-SR. Only in HT SS, but not in the other groups, salt restriction shifts the circadian rhythm of blood pressure from a non-dipper to a dipper pattern. We conclude that in black salt-sensitive hypertensives, salt restriction improves the circadian rhythm of blood pressure. This may have important therapeutic consequences on target organ damage associated with non-dipper patterns. PMID- 10804779 TI - [An evaluation of the impact of cardiovascular disease prevention in Portugal (1980-1996). A longitudinal Gompertzian analysis of mortality]. AB - The behaviour of cardiovascular diseases has suffered important modifications in recent years. There have been increases in mortality and morbidity in some countries while others have seen stabilization and even reduction. In the countries where we have observed a reduction in cardiovascular mortality, the reasons are multiple: efficacy of prevention measures; improvement of environmental conditions; increase in survival; and competitiveness with other causes of death. In order to ascertain whether a certain tendency is real or fictitious, we need to use techniques that allow the evaluation of the true evolution, to study the effect of environmental factors (not genetic) and the variability of internal resistance (vitality) to disease. Based on the longitudinal Gompertzian analysis of mortality for ischemic heart disease (IHD) and cerebrovascular disease (CVD) between 1980 and 1996, it was possible to observe a reduction in mortality for both causes, which was more obvious in the case of CHD, and that the impact of the improvement in environmental conditions (not genetic) was higher in CVD versus IHD. The prevention of CVD was more efficient in men (42.8% vs 29.7%) than in women, while in IHD the most efficient prevention was 20.6% in women versus 14.9% in men, comparing the data of 1996 and 1980. During this period, the environmental influences (not genetic) were responsible for the reduction in mortality for IHD in 2.7% of men and 4.3% of women, while in CVD we observed a reduction in mortality of 7.5% in men and 4.5% in women. PMID- 10804780 TI - [Stress-exercise echocardiography]. AB - The authors review the methodology and clinical applications of exercise echocardiography. This is a method that can be done in most of the patients with indication to the treadmill exercise test. There is an excellent relationship between costs and benefits associated to high sensitivities and specificities in the diagnosis of ischemia. This can lead to the preferential use of this technique, in the opinion of the authors, when it is indicated and in all patients that can do exercise. PMID- 10804781 TI - [A rare case of Marfan syndrome with involvement of the thoracoabdominal aorta- its surgical treatment]. AB - A 49-year-old woman with classic manifestations of Marfan's syndrome, having previously been submitted to two surgical procedures for ascending aorta and aortic valve replacement, was admitted with diagnosis of a large dissecting thoracoabdominal aortic aneurysm, with indication for surgical repair. The patient underwent a successful reconstruction of the aorta by means of the "simplified technique" introduced by ourselves in 1984, as an alternative to the conventional S. Crawford's "inclusion technique", for the management of thoracoabdominal aortic aneurysms. Assessed two years after the operation, the patient was found to be in excellent clinical condition and a control angiography showed the vascular prosthesis and its side branches working properly. This unusual aortic complication of Marfan's syndrome and the usefulness of the unique and original technique employed to replace the thoracoabdominal aorta justify the presentation of this clinical case. PMID- 10804782 TI - Tamponade as the clinical onset of a cardiac angiosarcoma. AB - Primary tumors of the heart are usually benign, with the malignant forms being rare. In this article we report the case of an angiosarcoma with the only manifestation being a cardiac tamponade. When the patient being admitted to the hospital, an important differential diagnosis was pulmonary tumor. A mass in the right atrium was later detected in the echocardiogram. Further image studies, such as MRI, showed spectacular images of this mass suggestive of neoplasm. An intra-operatory biopsy diagnosed the mass as an angiosarcoma. Since there was no evidence of primary neoplasm in other studies carried out, the neoformation was diagnosed as a primary angiosarcoma. Surgery was carried out with a survival of only 13 months after the procedure. We also use this case to make a brief review of the primary tumors of the heart, especially angiosarcomas. PMID- 10804783 TI - Right atrial thrombus treated with T-PA. PMID- 10804784 TI - Chest X-ray with large calcification of the pericardium--a clinical case. PMID- 10804785 TI - [Long-term benefit of primary angioplasty as compared with thrombolytic therapy for acute myocardial infarction]. PMID- 10804786 TI - [Epidemiology of drug-resistance and clinical microbiologists in the 21st century]. AB - Many efficacious antimicrobial agents have been developed in the latter half of the 20th century, and this has enabled us to overcome bacterial infections. However, various drug-resistant bacteria including MRSA and VRE have been emerging and these strains will cause serious life-threatening medical problems in the 21st century. Since development of new antimicrobial agents currently stagnates++, promoting the appropriate and prudent use of antimicrobial agents based on EBM is strongly recommended. Thus, establishment of a surveillance system of drug-resistant bacterial infections and their epidemiology as well as further activities by clinical technologists as specialists in medical microbiology will be indispensable in the next century. PMID- 10804787 TI - [Drug-resistant bacteria in clinical situations]. AB - There are two major categories of drug-resistant bacteria that can cause severe and intractable infections. The first includes multi drug-resistant Mycobacterium tuberculosis(MDRMT), penicillin-resistant Streptococcus pneumoniae(PRSP), and beta-lactamase positive or negative ampicillin-resistant Haemophilus influenzae, which used to be isolated from the patients with community-acquired infection. However, these pathogens have been often isolated in recent years from patients with hospital/chronic care facilities/nursing-home mass infection. The second major category includes methicillin-resistant Staphylococcus aureus(MRSA), vancomycin-resistant Enterococcus species(VRE), Pseudomonas aeruginosa, and Enterobacteriaceae including extended-spectrum beta-lactamases(ESBLs) producing strain, which are mainly isolated from compromised patients with nosocomial infections. The pathogenicity of these pathogens, almost all of which are found in the normal flora of humans, is weak, but often cause nosocomial infections in compromised patients. We need, therefore, surveillance system for these pathogens, and carefully determine whether these pathogens, if isolated, are causative pathogens. PMID- 10804788 TI - [Drug-resistant bacteria of current topics and their resistance mechanisms--PRSP and BLNAR]. AB - The prevalence of resistant isolates for beta-lactam antibiotics have been rapidly increasing in Streptococcus pneumoniae and Haemophilus influenzae. These strains having resistant elements are called penicillin-resistant S. pneumoniae(PRSP) and beta-lactamase-negative ampicillin-resistant H. influenzae(BLNAR). The mechanism of resistance was identified as the reduction of penicillin-binding proteins(PBPs) affinities as the target of beta-lactams. In PRSP, pbp1a, pbp2x, and pbp2b encoding PBP1A, -2X, and -2B enzymes, respectively, were altered. In contrast, mutations in the ftsI gene encoding PBP3A were only clarified in BLNAR. Characteristics of the resistance caused by PBPs alterations are a low-level resistance that decrease bactericidal action. PMID- 10804790 TI - [Drug-resistant bacteria of current topics and their resistance mechanisms- ESBL]. AB - Under the influence of antimicrobial agents, bacteria that primarily produce TEM Type or SHV-Type beta-lactamases developed point mutations in structural genes which served to extend the substrate specificity of the enzymes. Undoubtedly, CTX resistance rates increased in E. coli and K. pneumoniae in the recent years. It is strongly suggested that a survey of ESBL-producing clinical isolates of E. coli and K. pneumoniae should also be required for rigorous detection methods. PMID- 10804789 TI - [Drug-resistant bacteria of current topics and their resistance mechanisms--VRE]. AB - VRE isolates have only been detected in eight patients from four hospitals in Japan since 1996(reports of the Ministry of Health and Welfare, Japan). In Japan, it has been shown that VRE are high frequently isolated from chickens imported from Thailand and France where avoparcin has been used in animal feed. Two types of acquired glycopeptide resistance have been reported in clinical isolates of VRE. The VanA type strain is defined as having a high-level resistance to vancomycin and teicoplanin. The vanA resistance genes are induced by both vancomycin and teicoplanin. VanB type strains are characterized as having various levels of vancomycin resistance and are susceptible to teicoplanin. The vanB resistance genes are induced by vancomycin only. PMID- 10804791 TI - [Detection methods for drug-resistant bacteria in routine examination--MRSA]. AB - Since methicillin-resistant Staphylococcus aureus(MRSA) is resistant not only to methicillin but also to multiple species of antibacterial drugs, there are cases where infections with MRSA are difficult to treat. For laboratory identification of MRSA, S. aureus is first identified, then an oxacillin (MPIPC) sensitivity test, PCR detection of the S. aureus mecA gene and an agglutination test using the latex sensitized with an anti-PBP-2' monoclonal antibody are usually utilized. However, the detection of MRSA does not necessarily indicate MRSA infection, and many cases only demonstrate MRSA colonization. Thus, a clinical investigation is indispensable for establishing the diagnosis of MRSA infection. In addition, recent reports also suggested a current trend toward decreasing sensitivity to Bactroban(MUP, mupirocin, an anti-intranasal MRSA antibacterial drug). Taking these into consideration, surveillance of the trend in patient's sensitive to various species of antibacterial drugs including MUP must continue further. PMID- 10804792 TI - [The procedures for detection of penicillin-resistant Streptococcus pneumoniae and the epidemiology]. AB - Penicillin-resistant Streptococcus pneumoniae was first described in South Africa in 1977 and soon afterward was reported in other countries and in Japan. According to NCCLS standards for antimicrobial susceptibility tests, in vitro activity of S. pneumoniae is the best procedure using the minimum inhibitory concentration method. Benzylpenicillin, cefotaxime or ceftriaxone, and meropenem should be tested by a reliable MIC method and the susceptibility should be reported in the case isolated from blood and/or cerebrospinal fluid. Using the oxacillin test, it is difficult to distinguish penicillin-resistant from intermediate strains or sometimes susceptible strains. PMID- 10804793 TI - [Detection methods for drug-resistant bacteria in routine examination--VRE]. AB - When testing vancomycin susceptibility against enterococci, plates should be held a full 24 hours then examined using transmitted light. The presence of a haze or any growth within the zone of inhibition indicates resistance. Perform vancomycin MIC and tests for motility and pigment production to distinguish species with acquired resistance(vanA and vanB) from those with intrinsic, intermediate-level resistance to vancomycin(vanC) such as E. gallinarum, E. casseliflavus or E. flavescens. PMID- 10804794 TI - [Detection methods for drug-resistant bacteria in routine examination--ESBL]. AB - Extended-spectrum beta-lactamases(ESBLs) confer resistance to cefotaxime, ceftazidime, aztreonam, extended-spectrum penicillins, and structurally related beta-lactams in clinical isolates of K. pneumoniae and E. coli. Some ESBLs, however, show a lower level of resistance and these isolates may not reach NCCLS breakpoints for resistance. Optimal methods for confirmation of ESBL-producing isolates are described. PMID- 10804795 TI - [Detection methods for drug-resistant bacteria in routine examination--VISA]. AB - MRSA strains which clinically showed low-level resistance to vancomycin(VISA, hetero-VRSA) were isolated. Vancomycin-intermediate Staphylococcus aureus(VISA) was found to show a vancomycin minimum inhibitory concentration(MIC) of 8 to 16 micrograms/ml. The subpopulation of hetero vancomycin resistant MRSA(hetero-VRSA) was resistant to vancomycin MIC of 1 to 4 micrograms/ml. VISA and hetero-VRSA did not have vanA, vanB, vanC genes from vancomycin-resistant enterococci(VRE). The disk diffusion test could not detect VISA and hetero-VRSA. The MIC tests can confirm VRSA well. To detect the hetero-VRSA, we need to use population analysis and growth on hetero-VRSA agar (MU3 agar plate). PMID- 10804796 TI - [Detection methods for drug-resistant bacteria in routine examination--BLNAR]. AB - The characteristics of resistance, trend of prevalence and current status of beta lactamase-negative, ampicillin-intermediate or -resistant(BLNAR) strains of Haemophilus influenzae are summarized by our retrospectively analyzed data. The mechanism of this type of ampicillin-resistance is due to altered penicillin binding proteins(PBPs). Standardized susceptibility tests may differentiate ampicillin-resistant, beta-lactamase-producing strains from beta-lactamase negative, ampicillin-susceptible strains(BLNAS), but may not differentiate some strains of BLNAR from those of BLNAS. In the cases of serious infections, such as meningitis due to H. influenzae both beta-lactamase activity and ampicillin susceptibility should be confirmed by the MIC procedure. PMID- 10804797 TI - [Gene examination methods (detection and genotyping of resistant genes)--MRSA]. AB - Molecular diagnosis of MRSA infection including methods of detecting drag resistant gene and genotyping is described. In Japan, MRSA is the major pathogen for nosocomial infections. For symptomatic patients with MRSA infection, rapid detection of mecA and spa gene by PCR is quite critical to their prognosis. In cases of carry-over contamination of PCR products and mixed infection with coagulase-negative Staphylococci, take precautions are needed while working with this examination. Analysis by restriction length polymorphisms using pulsed-field gel electrophoresis (PFGE-RFLPs) for MRSA nosocomial infection is also described. PMID- 10804798 TI - [Gene examination methods (detection and genotyping of resistant genes)--VRE]. AB - There are six types of resistant genes in vancomycin-resistant enterococci: vanA, vanB, vanC-1, vanC-2, vanC-3, and vanD. The typing of the resistant genes is essential in clinical settings because the kind of a resistant gene implies susceptibility of an organism to glycopeptides and because strict infection control practices are required when vanA or vanB gene, which is encoded on plasmids and transmissible to other organisms, is detected. We describe methods of detecting and typing vancomycin-resistant genes, which are routinely performed in our laboratory. PMID- 10804799 TI - [Gene examination methods (detection and genotyping of resistant genes)--multiple drug-resistant Pseudomonas aeruginosa]. AB - Pseudomonas aeruginosa has been regarded as one of the most stubborn pathogens that easily acquire resistance to various antimicrobial agents. Recently, several clinical isolates that have acquired multiple-resistance to carbapenems, fluoroquinolones, and newly developed aminoglycosides such as amikacin have been reported. Thus, the "drug-resistant P. aeruginosa" was designated as a "class 4 pathogen" in the new "Law Concerning the Prevention of Infectious Diseases and Medical Care for Patients with Infections" in Japan. It is generally considered that no medical facility can escape from severe attack by both multiple-drug resistant gram-positive cocci such as MRSA and gram-negative rods including P. aeruginosa in the 21st century. Therefore, emergency countermeasures should be anticipated to prevent further proliferation of these multiple-drug resistant pathogens. PMID- 10804800 TI - [Gene examination methods (detection and genotyping of resistant genes)--metallo beta-lactamase]. AB - Metallo-beta-lactamases have a hydrolysing activity against various beta-lactams including carbapenems, and beta-lactamase inhibitors such as clavulanic acid and sulbactum. Infections caused by metallo-beta-lactamase producing gram-negative bacteria are becoming a serious problem in Japanese clinical settings despite the recent progress in medical technology. Current screening methods can not distinguish metallo-beta-lactamase producers from those producing AmpC beta lactamase. Here we show a convenient method of detecting metallo-beta-lactamase producers using a thiol compound(2-mercaptopropionic acid) as well as a PCR method with metallo-beta-lactamase gene specific primers. PMID- 10804801 TI - [Infections with drug resistant bacteria and their treatment methods--MRSA infections]. AB - MRSA infection is difficult to treat because of the pathogenecity of causative strains and their resistance to many kinds of antibiotics not all strains of MRSA are pathogenic agents and indications for antibiotic use should be determined strictly. Although several antibiotics show bactericidal activity against MRSA, they are not effective in all patients infected with MRSA. We should select the best drug according to the characteristics of the respective drugs in each patient and search for a better regimen to treat severely infected cases. PMID- 10804802 TI - [Infections with drug resistant bacteria and their treatment methods--PRSP infections]. AB - S. pneumoniae is one of the most important pathogens in respiratory tract infections and systemic infections. The frequencies of isolating Penicillin resistant Streptococcus pneumoniae(PRSP) has been increasing recently. The therapeutic strategy is becoming difficult. Ampicillin and amoxicillin are the choices of drugs to treat respiratory infections with intermediate strains. Panipenem may be the first choice of treatment for systemic infections. Administration of vancomycin should also be considered. Judicious use of antimicrobials is necessary to avoid the spread of multi-resistant strains. In some cases physical therapy would be recommended. PMID- 10804803 TI - [Infections with drug resistant bacteria and their treatment methods--VRE infections]. AB - Vancomycin-resistant enterococci(VRE) is a worldwide threat now. Because we have lost a last resort vancomycin for enterococcal infections, and their resistant genes can be transferred to other more pathogenic gram-positive bacteria. We have only a few of optional therapeutic agents against VRE. VRE causes urinary tract infections, intra-abdominal infections, bacteremias, endocarditis and wound infections. Especially bacteremias caused by VRE result in high mortality. VRE infections including colonization and infectious diseases must be controlled by rapid detection of VRE, appropriate diagnosis of infections, followed by effective antimicrobial therapies and infection control measures. PMID- 10804804 TI - [Quality control on antimicrobial susceptibility testings]. AB - Performance of quality control in the clinical microbiological laboratory is markedly behind that of other laboratories. In particular, it remains indispensable even if the antimicrobial susceptibility test being carried out is standardized. In Japan, a standardized method of antimicrobial susceptibility testing has recently been established by the National Committee for Clinical Laboratory Standards and Japan Society of Chemotherapy. However, most laboratories do not perform quality control for antimicrobial susceptibility testing according to the standardized rules. The problem in quality control in laboratories causes things to remain as they are. As a review of laboratory quality control methods, we will show several cases of error that we encountered in our laboratory. PMID- 10804805 TI - [The present status of susceptibility tests for detection of antibiotic-resistant bacteria in Japan]. AB - Recently, NCCLS Susceptibility tests have been widely used in Japan. NCCLS Susceptibility tests are one of the most excellent systems in the world. However, there are some differences in the type of antibiotic-resistant bacteria and systems of chemotherapy between Japan and the USA. To use NCCLS methods more effectively in Japan, it is necessary to make a partial amendment concerning with the choice of antibiotics and break points of resistance. PMID- 10804806 TI - [Proceeding of the workshop "What an ideal clinical microbiological laboratory should be". The 33rd in-service training course for medical technologists of university hospitals in 1999]. AB - In the workshop of the 33rd in-service training course for University Hospital Medical Technologists in 1999(sponsorship: the ministry of Education), all groups of participants were charged with discussing an ideal clinical microbiology laboratory. In conclusion, the successful operation of the ideal system of clinical microbiology should require a high level of competence in every staff member of the hospital. It must not be focused solely on the sophistication of laboratory methods. We must modify our behavior effectively and establish a good collaborative partnership with physicians and other health care professionals. PMID- 10804807 TI - ["What an ideal clinical microbiological laboratory should be"--from the position of laboratory manager]. AB - The most important thing for clinical laboratory is the "labor-power". "Labor power" involves knowledge, technique, activity, analysis, patience and above all else benevolence. All of these constitute an efficiency examination(that is a total analysis of necessity, rapidity, correctness, simplicity, cost). PMID- 10804808 TI - ["What an ideal clinical microbiological laboratory should be"--from the position of medical technologist]. AB - The evolution of the microbiology laboratory is necessary for correspondence to the transfiguration of infection and contribution to clinical applications. Especially, the correspondence of emergency tests such as smear strain and antigen detection, the report added value and the infection surveillance in team medical treatment are indispensable. Also, medical technologists need to be knowledge able about techniques related to infection overall, and participation in infection diagnosis and social responsibility are indispensable. PMID- 10804809 TI - ["What an ideal clinical microbiological laboratory should be"--from the position of patient]. AB - To think about the ideal clinical microbiological laboratory in patient's place, there are three important problems in present medical treatment. The first one, for patients, it is necessary to wait many hours to take a medical advice from a doctor. The second, patients should be checked many examinations. The third, the heavy patient's share in his medical expenses. To settle a bit these matters, the clinical laboratory should try to accept the rapid and economical examinations for patients. PMID- 10804810 TI - [The new law concerning the prevention of infectious diseases and medical care for patients of infections]. AB - A new law concerning the prevention of infectious diseases and medical care for patients with infectious diseases was enacted on Oct 2, 1998 and became effective on April 1, 1999. In the new infection control law, all infectious diseases are included as the targets of national epidemiological surveillance and are classified into new categories 1 through 4, depending on their contagiousness and severity. Under this new law, the national epidemiological surveillance of infectious diseases is defined as one of the main objects. This new law also specified the provision of a medical care supply system including assigned hospitals and medical fees. PMID- 10804811 TI - [Revolution of the health care delivery system and its impacts on laboratory testing in the United States]. AB - Failure to slow the exponential growth of total health care expenditures in the United States through the government policies resulted in a rapid and progressive penetration of managed care organizations(MCOs) in the early 1990s. Diagnostic testing is viewed as a "commodity" rather than a medical service under the managed care environment. Traditional hospital-based laboratories are placed in a downward spiral with the advent of managed care era. A massive reduction of in house testing resulted from shorter lengths of patients' hospital stay and a marked decrease in admission under the dominance of managed care urges them to develop strategies for restoring tests deprived by the managed care-associated new businesses: consolidation and networking, participation in the outreach testing market, and point-of-care/satellite laboratory testing in non traditional, ambulatory settings are major strategies for survival of hospital laboratories. A number of physicians' office laboratories(POLs) have been closed owing to regulatory restrictions imposed by the Clinical Laboratory Improvement Amendments of 1988(CLIA '88), and to the expanded penetration of MCOs which limit reimbursement to a very few in-house procedures. It seems likely that POLs and hospital laboratories continue to reduce test volumes, while commercial reference laboratories(CRLs) gain more tests through contracting with MCOs. In the current stream of managed care dominance in the United States, clinical laboratories are changing their basic operation focus and mission in response to the aggressively changing landscape. Traditional laboratories which are unwilling to adapt themselves to the new environment will not survive in this country. PMID- 10804812 TI - [Progress in biochemical and genetic examinations in clinical laboratory of cardiovascular diseases]. AB - For diagnosing cardiovascular emergencies, cardiologists mainly rely on physiological examinations. Biochemical laboratory examinations can facilitate emergency diagnosis and genetic diagnosis can facilitate a fundamental understanding of cardiovascular diseases. Recent progress in biochemical examinations for cardiovascular diseases includes measurement of troponin T, heart type fatty acid-binding protein, and LDL-cholesterol. Recent progress in genetic examinations includes analysis of mitochondrial DNA, obesity-related genes and single nucleotide polymorphisms. A safe and convenient system of collecting, transporting, and storing blood and other biological specimens, and the subsequent purification of genomic DNA from the stored specimens is available. Therefore, busy doctors and staff of clinical laboratory can plan subsequent DNA examinations without causing concern and pain. A recently reported human mutation of heterotrimeric G protein beta 3 subunit produces a gain-of function G protein signaling abnormality. A common polymorphic base substitution, C825T, in the GNB3 gene is associated with an aberrantly spliced transcript lacking 123 nucleotides. A significant association of the T allele with essential hypertension or obesity has been suggested. PMID- 10804813 TI - [Stress testing in cardiology and cardiac rehabilitation]. AB - In recent years, investigations into cardiac rehabilitation have suggested that bicycle exercise training increases peak oxygen uptake of patients with chronic heart failure(CHF). However, some patients can not perform such exercise because of poor cardiac function. If patients were able to achieve metabolic improvement in their muscles by localized small muscle group training, it would be advantageous for these patients. Therefore, in patients with CHF, we investigated whether localized skeletal muscle training improved calf muscle metabolism. Seven patients undertook a random order crossover trial. Training consisted of unilateral calf plantar flexion exercise. After training, phosphorus-31 nuclear magnetic resonance spectroscopy revealed significant improvements in calf muscle metabolism in patients with CHF. Subjective fatigue score was also improved. In this paper, stress testing in cardiology and cardiac rehabilitation including the above experimental data were introduced. PMID- 10804814 TI - [A role of nuclear medicine in diagnosing cardiac disease--clinical use of 123I BMIPP and 123I-MIBG]. AB - There are many new nuclear methods in diagnosing cardiovascular disease. We present about 2 nuclear diagnosing methods. One is about 123I-BMIPP and the other 123I-MIBG. 123I-beta-methyl iodophenyl pentadecanoic acid(123I-BMIPP) is the tracer to evaluate the metabolism of a fatty acid of the heart. A combination of perfusion and fatty acid imagings indicates the recovery of cardiac wall motion can be expected in acute myocardial infarction in zones where 123I-BMIPP decreased uptake is greater than 201T1 perfusion defect. In angina pectoris, especially in unstable angina, 123I-BMIPP may be available to detect myocardial ischemia. 123I-meta-iodobenzylguanidine(123I-MIBG) is the trace that be able to assess the heart with sympathetic innervation. 123I-MIBG scintigraphy is a useful noninvasive method for the assessment of not only myocardial ischemia but also the degree of congestive heart failure. Also, it is a useful in assessment of beta-blocker therapy. PMID- 10804815 TI - [Recent advances in echocardiographic diagnosis of ischemic heart disease]. AB - Before the early 1990s, the diagnostic usefulness of echocardiography for ischemic heart disease had been relatively limited compared with that for other cardiac diseases such as valvular disease, congenital anomalies and cardiomyopathies. The principal role of echocardiography was to assess persistent regional wall motion abnormality as well as to detect complications of myocardial infarction. However, recent technological advances have created many newer applications of echocardiography in this field. One of the most important advances was seen in the field of stress echocardiography. Dobutamine stress echocardiography has become an established method of diagnosing transient myocardial ischemia due to coronary stenosis and assessing the myocardial viability of a persistently akinetic segment. More recently, several new contrast agents have been developed or will be available in the near future to visualize the blood stream within the left heart cavity and myocardial blood flow. Simultaneously, new ultrasound technologies including harmonic imaging and gated intermittent imaging have enhanced the selective visualization of contrast agents and will contribute to noninvasive imaging of coronary microcirculation. Harmonic imaging has also been shown to improve quality of B-mode image without a contrast agent and will play an important role in the clinical recognition of wall motion abnormality in patients with ischemic heart disease. Recent advances in three dimensional technology have enabled accurate measurements of left ventricular volume and ejection fraction without any geometrical assumption, which may be especially important for the evaluation of ischemic patients who often have a deformed left ventricular cavity due to remodeling. PMID- 10804816 TI - [Invasive examination of cardiovascular disease]. AB - Invasive cardiovascular examination by coronary angioscopy and by using a Doppler guide wire, a pressure guide wire and intravascular ultrasound has extensively developed. A Doppler guide wire is used for measurement of flow velocity and evaluation of coronary blood flow. Previous studies demonstrated by assessing maximum coronary vasodilatory capacity that endothelium-dependent or independent vasodilation was impaired in hypertension and hypercholesterolemia or in syndrome X. Elevation of coronary vascular resistance during coronary microvascular spasm has been verified by using a Doppler wire. A pressure guide wire provides coronary transstenotic pressure and is available in calculating myocardial fractional flow reserve(FFRmyo). FFRmyo is an important parameter to assess the coronary functional stenosis that is culprit for myocardial ischemia. It is calculated from the ratio of the mean transstenotic pressure to the mean pressure proximal to the stenosis during maximum coronary hyperemia. The value of FFRmyo considered as necessary for coronary intervention is below 0.75. Intravascular ultrasound(IVUS) is applied as both a diagnostic tool and for intervention purpose. It enabled tissue characterization of the vascular wall as well as measurements of vascular diameter, vascular lumen area and plaque area. It also aided in optimal devise selection, decision of interventional endpoint and assessment of restenosis. IVUS promoted deployment of high-pressure stents to obtain a large post-procedural lumen area as well as abolition of anticoagulation in case of optimal stent deployment. Coronary angioscopy has been developed to investigate the pathogenesis of acute coronary syndrome, where disrupted yellow plaque and overlying thrombus play important roles. Angioscopy has also evidenced regression of intimal hyperplasia after coronary stenting. PMID- 10804817 TI - [Genetic diagnosis for cancer--tumorigenesis and genetic alterations]. AB - During malignant transformation, cancer cells acquire multiple genetic alterations that override the normal mechanisms controlling cellular proliferation. In brief, cancer is a disease of genetic abnormalities caused by hereditary and/or environmental factors. Genetic diagnosis for cancer can be divided into four categories: 1) pre-symptomatic diagnosis, 2) existence diagnosis, 3) property diagnosis-prognosis diagnosis and 4) genetic test for gene therapy. 1) For hereditary cancer families, pre-symptomatic diagnosis is available. Individuals with multiple cancers can be diagnosed by microsatellite instability(MSI) test using resected cancer tissues and by genotyping of mismatch repair enzymes. If the genotype abnormality is detected, the propositus can obtain early diagnosis and prevention of cancer, and genetic services of their siblings. 2) Using PCR technology, occult tumor cells can be detected from blood and other biological body fluids, as targets of chimeric transcripts and tissue specific expressions. 3) Molecular properties of cancer cells are investigated for grading malignancy and therapeutic sensitivities. Using the molecular properties, prognosis of the patient can be estimated. 4) Following the genetic test results, the specific and superior gene therapy can be applied individually. Post-therapeutic monitoring is also available only by genetic test. After the genome project, the significance of single nucleotide polymorphisms(SNPs) related with cancer will be established, then tailor-made therapy and/or prevention will be applicable to individuals. The ultimate goal of genetic diagnosis would be the final priority to the phenotypic diagnosis. PMID- 10804818 TI - [Molecular-based diagnosis of thyroid carcinomas by detecting cancer-specific mRNAs]. AB - Molecular-based diagnosis of thyroid carcinomas can be more easily established by utilizing specific mRNAs that are restrictedly expressed in cancer tissues. In light of this aspect, we searched for cancer-specific mRNAs using sequence specific-differential display(SS-DD) and serial analysis of gene expression(SAGE). By these techniques, we found several mRNAs that efficiently distinguished benign and malignant tissues. Among these, oncofetal fibronectin(onfFN) mRNA was expressed only in thyroid papillary and anaplastic carcinomas and it was considered the most preferable target for molecular-based diagnosis of these carcinomas. In a previous study, we introduced a new method of preoperative diagnosing thyroid carcinomas. This technique, aspiration biopsy-RT PCR(ABRP), allows us to simultaneously perform cytological and molecular-based diagnoses by extracting RNA from leftover cells within the needle used for fine needle aspiration biopsies(FNABs). ABRP provides both RNA information and cytological diagnosis without further invasion to the patient. We demonstrated that by ABRP detection of onfFN mRNA in FNABs, papillary and anaplastic carcinomas may be accurately diagnosed preoperatively. Further, by real-time monitoring RT-PCR measurement of onfFN mRNA, a fully automated system was established. PMID- 10804819 TI - [Genetic diagnosis of leukemia: diagnosis of relapse and complete remission, and prediction of leukemia onset]. AB - Wilms' tumor gene WT1 mRNA is a new marker of leukemic blast cells for AML, ALL, and CML. Minimal residual disease(MRD) of leukemia can be detected at frequencies as low as 1 in 10(3) to 10(4) normal bone marrow cells and 1 in 10(5) normal peripheral blood mononuclear cells by means of the quantitation of WT1 mRNA(WT1 assay) using reverse transcriptase-polymerase chain reaction. Thus, the WT1 assay makes it possible to rapidly assess the effectiveness of treatment and to evaluate the degree of eradication of leukemic cell in individual leukemia patients. Furthermore, WT1 assay can continuously assess the disease progression of myelodysplastic syndromes(MDS) and predict the evolution of MDS to overt AML within 6 months. PMID- 10804820 TI - [Genetic diagnosis for drug resistance in cancers]. AB - The failure of chemotherapy to eradicate tumor cells is often due to the development of drug resistance. MDR(multidrug resistance) whose one form of resistance results from a decreased intracellular accumulation of the drugs, most often mediated by the overexpression of P-glycoprotein. MRP also related to pump function of cell membrane in acute leukemia. We have developed the new quantitative assay based on real-time PCR to measure expression of drug resistance related genes such as MDR-1 and MRP in clinical samples. These results indicates that real-time PCR system is a reliable method to quantitatively determine drug resistant genes expression, it may be to predict responsiveness to chemotherapy by using this technique. PMID- 10804821 TI - [Antimicrobial susceptibility testing for Mycobacterium tuberculosis by the bioluminescence assay of mycobacterial ATP using filamentous cell treatment]. AB - The antimicrobial susceptibility testing for Mycobacterium tuberculosis by the bioluminescence assay of adenosine triphosphate(ATP) derived from living mycobacteria was improved introducing filamentous cell treatment(FCT) reported for beta-lactam susceptibility test of Pseudomonas aeruginosa by Hattori. Before ATP extraction, bacterial cells were treated with the FCT reagent for 30 minutes at room temperature. Adenosine phosphate deaminase in the FCT reagent simultaneously digested the extracted ATP and released ATP in a liquid culture of M. tuberculosis H37Rv and the RLU level was decreased markedly. Using this improved ATP method, we determined the ATP contents of M. tuberculosis inoculated into Middle-brook 7H9 broth medium with or without drugs. In ethambutol(EB) susceptibility, the ATP method reported previously, showed false-resistance when judged within 7 days. To eliminate false-resistance in EB susceptibility we applied the modified ATP method with FCT treatment to strains determined EB susceptible by reference methods. Using this modified ATP method, we could judge EB susceptibility of 5 ATCC reference strains within 3 days, and these of 15 clinical isolates of M. tuberculosis within 5 days. And all the results obtained were coincident between the ATP method and the reference methods. The reproducibility of this modified ATP method was evaluated with six ATCC reference strains at the concentrations of 0.1 microgram/ml of isoniazid(INH), 2.0 micrograms/ml of rifampicin(RFP), 2.5 micrograms/ml of EB, 2.0 micrograms/ml of streptomycin(SM), and 5.0 micrograms/ml of kanamycin(KM). The test was repeated six times. Reduction of ATP contents were observed in susceptible strains but not in resistant ones within 3 days of cultivation and susceptibilities to drugs could be determined within 3 days at every time when combined FCT to the ATP method. And highly reproducible results were obtained. It is strongly suggested that this modified method is simple, rapid, highly reproducible and nonradiometric, and could be used for the assessment of drug susceptibility for M. tuberculosis. PMID- 10804822 TI - [No significant influence of serum amyloid A1 genotypes on serum lipids or HDL clearance]. AB - Serum amyloid A1(SAA1), the major acute phase isotype of SAA protein family, consists of three common allelic variants(SAA1.1, SAA1.3 and SAA1.5) in the Japanese population. We have recently reported that subjects with the SAA1.5 allele have higher plasma SAA concentrations than those without it, a phenomenon probably due to the delayed catabolism of the isotype SAA1.5. Since SAA is present in high density lipoprotein(HDL), this study assessed whether SAA genotype influenced the serum lipid study by altering HDL metabolism. In a total of 279 healthy adults, no difference was noted in their total cholesterol, HDL cholesterol or triglyceride concentrations among six genotype groups. Plasma clearance of human apolipoprotein AI(apoAI) was studied in mice by giving HDL reconstituted with each recombinant human SAA1 isotype. The apoAI clearance did not differ among each of the SAA1 isotype-conjugated HDLs. Moreover, the changes in content of SAA in HDL also did not alter the apoAI clearance. These results suggest that SAA1 may not play an active role in plasma HDL metabolism. PMID- 10804823 TI - [A luminescent assay for natural cytotoxicity using K562 cells transfected with the plasmid producing alkaline phosphatase]. AB - A novel non-radioactive assay for natural cytotoxicity is described. A new target cell line was established by the transfection of K562 cells with a plasmid encoding DNA for secretion type of placental alkaline phosphatase(ALP). The assay was based on the kinetic determination of heat-stable ALP in the co-culture of the target cells with effector sample from human peripheral blood. The inactivated effector sample for the control prepared by irradiation(50Gy X-ray) or heat treatment(56 degrees C, 20 min), however, showed an enhancement effect on the ALP production in the target cells. This effect was depending on the concentration of monocyte component(adherent cells on plastic dish) by unknown reason, and was not reduced by the treatment for NK inactivation. Cytotoxic activity was therefore determined by the percent decrease in the ALP productivity compared to that in the control. The correlation coefficient between cytotoxicity values by this ALP synthesis assay and those by 51Cr release assay was 0.93. This cytotoxicity test is simple and unique in terms of estimation for protein synthesis affected in target tumor cells, although the mechanism of the monocyte(or macrophage) effect remained to be elucidated. PMID- 10804824 TI - [Hepatitis B virus gene mutations in the sera of three patients with coexisting hepatitis B surface antigen and anti-surface antibody]. AB - We analyzed gene mutations in the Hepatitis B virus of three virus carriers with coexisting Hepatitis B surface(HBs) antigen and anti-HBs antibody. Viral DNAs were extracted from sera and the pre-S, S and X(including core promoter and pre core region) regions were amplified by PCR, and sequenced. Case 1 and Case 2 were positive for HBe antigen, while Case 3 was negative. All three cases were positive for HBe antibody and HBV DNA. In the S gene region, various point mutations were detected in all three cases. Mutations were clustered in the first hydrophilic loop region(codon 47-46) essential for the secretion of surface antigen. A few mutations were detected in 'a' loop(codon 124-147) of the S gene. None of the cases had an amino acid substitution of codon 145 of the S gene that is reported to be responsible for weak recognition by the HBs antibody. These data suggest the existence of hyper-variable sequence in S region, or otherwise result of low-fidelity of Taq DNA polymerase-reaction. Case 1 possessed a point mutation, T to C at nucleotide position 1753, in the region overlapping the coding region of the X gene and the CCAAT/enhancer binding protein(C/EBP) binding region within the core promoter region. Case 2 possessed both a large deletion(129 bp) in the pre-S1 and in-frame deletions of 15 and 27 bp in the pre S2 region. Case 3 had an in-frame deletion of 30 bp in the pre-S2 region, and a point mutation in precore region. The point mutation, G to A at a nucleotide position 1986, converts Trp(TGG) to a stop codon TAG, and may contribute the fulminant hepatitis. These results suggest that the mutations in the pre-S, the core promoter, or the X gene may imply coexistence of the HBs antigen and antibody after seroconversion, while the point mutations in the S region are not likely to be responsible for the HBV escape mutant. PMID- 10804825 TI - [Evidence-based laboratory medicine--a new trend in laboratory medicine]. AB - The most important target of "evidence-based laboratory medicine(EBLM)", based on the fundamental concept of evidence-based medicine(EBM) is to improve appropriate, effective utilization of laboratory tests through close communication between clinical laboratories and clinicians. It is mandatory for clinicians to know of analytical uncertainty for better utilization of laboratory tests in clinical practice. Furthermore, the improvement of clinical utilization and interpretation of laboratory tests can be expected by supplying the evidences obtained through systematic reviewing or meta-analysis of laboratory tests. In order to pursue these purposes, closer communication and cooperation between laboratories and clinicians are important to obtain an effective consensus. Clinical laboratory must work together with interested clinician(s) for systematic review of laboratory tests. Construction of the ways of better laboratory-clinician communication should be an important paradigm for EBLM which will be an important factor for new revolution of laboratory medicine in future. PMID- 10804826 TI - Current status of clinical laboratory in Mongolia. PMID- 10804827 TI - [Reference values for hematologic laboratory tests and hematologic disorders in the aged]. AB - The establishment of reference values in the complete blood count and lymphocyte subsets for the aged is necessary for diagnosis and therapy since the number of the aged is increasing rapidly in recent years. However, it is problematical to define the healthy aged because of physiological changes with aging, compared with the healthy adults. Red blood cell count, hemoglobin concentration(Hb) and hematocrit for the healthy aged showed the tendency of moderate decreasing with aging. These phenomena become more obvious by dividing the healthy aged into three groups, 65-74 years, 75-84 years and over 85 years. The mean corpuscular volume(MCV) for the healthy aged showed the tendency of increasing with aging, while mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration for the healthy aged showed no tendency with aging. No changes for the healthy aged were observed in the leukocyte differential, white blood cell count and platelet count with aging. Results of lymphocyte subsets(CD3, CD4, CD8, CD19 and CD4/CD8 ratio) for the healthy aged are not coincident among reporters at present because the healthy aged population is different and the determination of lymphocyte subsets is not standardized. It is important to diagnose as anemia below Hb 11.0 g/dL between 65 and 74 years, Hb 10.0 g/dL over 75 years using Hb level with high accuracy. Most of hematologic disorders in the aged are the secondary anemia based on malignant tumor. In the aged between MCV 95 and 120 fL or over MCV 120 fL, myelodysplastic syndrome or vitamin B12 deficiency is considered, respectively. In the future, more vertical study in the aged population is needed to establish the reference values for the aged. PMID- 10804828 TI - [Coagulation/fibrinolysis disorders in the elderly]. AB - To establish reference intervals for the interpretation of hemostatic tests in the elderly, 11 hemostatic tests were performed on 120 elderly individuals(26 males and 94 females) aged 78.6 years in average. The subjects lived in hostel for the elderly, walked without assistance, and were independent in activities in daily living. In 25.8% to 93.3% of the subjects, the results of the tests, except for prothrombin time and thrombin-antithrombin III complex, deviated from reference intervals obtained from young healthy subjects. This suggests the necessity of setting up reference intervals for the elderly. Fibrinolysis was more activated than coagulation, although coagulation had been considered to be more activated because thrombosis is common in the elderly. Wilcoxon test on the test values of the early elderly(65 to 74 years of age) and late elderly(75 to 92 years of age) as well as the correlation coefficient for age and test values revealed that separate reference intervals for antithrombin III, protein C, D dimer, and thrombomodulin should be established for the early elderly(65 to 74 years) and late elderly(75 years or over). Our proposed reference intervals for the elderly are appropriate for the diagnosis of disseminated intravascular coagulation. PMID- 10804829 TI - [Decision levels of diabetes mellitus and hyperlipidemia in elderly Japanese subjects]. AB - Decision levels of diabetes mellitus and hyperlipidemia in elderly subjects as well as younger subjects should be determined to prevent complications of these diseases such as microvascular and macrovascular diseases. Therefore, prospective follow-up study is necessary to decide the decision levels. In the case of diabetes mellitus, there are some useful studies such as KUMAMOTO STUDY show that strict glycemic control can prevent microvascular diseases, but there are few studies in elderly subjects. However, hypoglycemia causes macrovascular events, and chronic hypoglycemia leads to dementia and apathy. It is generally accepted that the glycemic control level can be milder than that in younger subjects. We suggest that the following decision levels are reasonable for elderly diabetic subjects, 1. FPG > 140-160 mg/dl, 2. PG(2 h) > 200-250 mg/dl, 3. HbA1c > 7-8%. Decision level of hyperlipidemia in elderly subjects should also be determined to prevent cardiovascular disease. It is demonstrated that anti-hyperlipidemic treatment can prevent CHD even in elderly subjects by many prospective studies. Japan Atherosclerosis Society recommend that the decision levels of hyperlipidemia in elderly subjects can be the same as younger subjects. The decision levels indicating diet therapy and medication for risk factor free subjects(category A) are LDL-C > or = 140 and 160 mg/dl, respectively. Those for subjects without CHD but have some risk factors(category B) are LDL-C > or = 120 and 140 mg/dl, respectively. Those for subjects with CHD(category C) are LDL-C > or = 100 and 120 mg/dl, respectively. PMID- 10804830 TI - [Liver function of the most elderly patients]. AB - To determine the decision level of liver function in the most elderly patients, we compared serum albumin, aspartate aminotransferase(AST), alanine aminotransferase(ALT) and alkaline phosphatase(ALP) values of the most elderly patients > 85 years with those of healthy young adults. Two hundred fifty five elderly people, aged 88-106 years and average 96.6 years(171 women, 84 men), were included in this study. Elderly people were divided into four groups according to their activities of daily living(ADL), 114 Rank-J: free living, 62 Rank-A: unable to go outside without help, 39 Rank-B: bedridden but able to sit up in bed and 40 Rank-C: completely bedridden. Serum albumin values for the most elderly patients in Rank-J were 4.2 +/- 0.3 g/dl for women and 4.0 +/- 0.3 g/dl for men, showing marked decrease from those of young healthy adults aged 19-59 years(p < 0.0001). In 22.2% of elderly women and 44.2% of elderly men, albumin values deviated from the reference interval of young adults. ALT value for the most elderly patients also showed a decrease in both sexes and AST and ALP values for the most elderly patients showed an increase in women compared with young adults. However, these were minor deviations from the reference interval for young adults. In ADL stratified groups of the most elderly patients, serum albumin values showed marked decrease with decline in ADL, whereas AST, ALT and ALP values remained constant in both sexes regardless of ADL. PMID- 10804831 TI - [Endocrine diseases and aging]. AB - Age dependent perturbations to endocrine systems are overviewed and the interpretation of the results of endocrinological tests for clinical decision are discussed referring to thyroid diseases as an example. The endocrine functions that are essential to life, such as adrenal and thyroid functions, show minimal change by aging. Their basal levels alter little, although production and degradation of hormones decrease. Some part of elderly persons show lower levels of triiodothyronine, which may, however, reflect deterioration of their physical condition and should not be overlooked as an age dependent change. Some endocrine function, such as gonadal function and calcium metabolism obviously reveal age dependent physiological change and their reference ranges for elderly persons are set separately. Thus, reference levels and clinical decision for endocrinological tests should be dependent on the properties of individual endocrine system. PMID- 10804832 TI - [Decision levels of sero-immunological data]. AB - OBJECTIVES: To determine reference values in several representative serological or immunological examinations in the elderly subjects compared with young adults. MATERIALS AND METHODS: The group of elderly subjects over 70 years consisted of 630 people living at their own home and 170 people living in nursing homes. The young control group consisted of the 340 university students. ASO and RF were quantitatively measured by latex turbidimetry and FANA by immunofluorescence. Furthermore, several other autoantibodies were also examined by ELISA. The ninety fifth percentile method was employed for the determination of reference values. RESULTS: The upper limit of reference intervals for ASO was 190 U/ml in the elderly group, while 330 U/ml in the young group. As to RF, it was 25 U/ml (70-79 years) and 37 U/ml (80-89 years), while 15 U/ml in the young. In FANA, x 160 was determined in both the elderly and the young adult subjects. Relatively higher positive frequencies were obtained in the autoantibodies to desmogrein3, dsDNA, cardiolipin and thyroglobulin in the elderly. CONCLUSION: In the elderly, ASO titers were lower, RF titers were higher, and FANA titers were generally similar to those in young adults. Higher titers were observed for some other autoantibodies. PMID- 10804833 TI - [Outsourcing of clinical laboratory department]. AB - Recently, to improve financial difficulties at various hospitals, outsourcing of the laboratory department is be coming more wide spread. At the department of clinical pathology of St. Luke's International Hospital, the system, so called, "Branch labo" which is one of the outsourcing laboratory conditions, was adopted in March 1999. In this reports. We described the decision procedure for accepting the situation and the circumstances of operation. PMID- 10804834 TI - [A report from Committee on Essential Laboratory Tests for initial Diagnosis of Patients]. AB - Recently, medical circumstance is changing and fixed-fee reimbursement systems such as DRG/PPS are now going to be established in our country. Last year, the Japan Society of Clinical Pathology(JSCP) newly set up the Committee on Essential Laboratory Tests for Initial Diagnosis of Patients to keep up with current trends of medical care environment. In this Committee, we have discussed how to ensure the minimum requirement of laboratory tests in patients when they have or may have one of the disease of DRG. And finally the Committee released the proposed guideline for usefulness of the essential laboratory tests in 9 disorders that were randomly picked from DRG. The guidelines were then delivered to members of JSCP, or other medical associations, and doctors working at hospitals in which DRG/PPS system were on going as a trial. We have had many opinions or criticisms from doctors who received the guideline. Now the Committee is planning to publish next version of guideline in which we will have more 14 disorders from DRG and have modifications of original version. PMID- 10804835 TI - [A trial of prospective payment system (DRGs/PPS) in Sendai National Hospital]. AB - This study was undertaken to compare the difference between the total payment that was actually paid in accordance with the rules of DRGs/PPS, and the medical fee calculated by the present Japanese medical insurance system. From Nov. 1st '98 to Oct. 31st '99, the DRGs/PPS was applied to the 1,723(21.4%) out of 8,042 acute inpatients who were discharged after short-term stay. The total payment by the DRGs/PPS(yen 1,450,725,370) was smaller than the medical fee by the present Japanese insurance system(yen 1,504,686,240). In MDC (Major Diagnostic Category) 1(nervous system), 2(eye), 3(ear, nose, throat) and 4(circulatory system) the payment by the DRGs/PPS was greater than the fee calculated by the present medical insurance system. On the other hand, patients who belonged to the other 9 MDC groups paid smaller amounts by the DRGs/PPS. The major factors accounting for the difference(yen 53,960,870) between total payment DRGs/PPS and the fee by the present insurance system were period of hospital stay, treatment in the intensive care unit, complication, cancer chemotherapy, expensive surgical materials etc. PMID- 10804836 TI - [A cost-effectiveness analysis of a pre-visit testing system for new outpatients in primary care medicine]. AB - We evaluated cost-effectiveness parameters of common diagnostic test panels advocated by the Japan Society of Clinical Pathology in order to determine the validity of pre-visit testing of these test panels for new outpatients with some defined symptoms. The "Essential Laboratory Tests(ELT) (2)" panel was universally applied to 540 new outpatients who visited the Comprehensive Medicine Clinics in the National Defense Medical College and its affiliated hospital. A "useful result(UR)", the unit of usefulness of the ELT, was assigned by assessing the impact of its results on physician's diagnosis- or decision-making. Patients were classified into 7 major symptom groups according to their chief complaint, and clinical effectiveness(UR/patient), economic efficiency(cost/UR) and cost effectiveness(delta cost/delta UR) were analyzed in each patient group. Clinical effectiveness ranged from 1.38(renal/urinary tract symptoms) to 0.27 UR/patient(neurological symptoms) with a cost disparity from yen 1,222 to yen 5,946/UR between these patient groups. Substantial decreases in clinical effectiveness were demonstrated in all patient groups with the ELT(1)-testing alone in a simulation study; however, cost-effectiveness parameters still remain in excellent values in the renal/urinary tract symptom and infection/inflammation related symptom groups(1.13 and 0.77 UR/patient at a cost of yen 836 and yen 1,181/UR, respectively). The addition of some ELT(2)-specific test items such as AST, ALT to the ELT(1) baseline panel improved clinical effectiveness of the ELT(1) in lassitude/exhaustive symptom group at a minimal cost increment. This study verified the validity of the ELT(1) as a panel for pre-visit testing in patients with renal/urinary tract symptoms and infection/inflammation-related symptoms, but not in those with other symptoms. PMID- 10804837 TI - [Studies on circulating interleukin-6 and thyroid functions in acute myocardial infarction]. AB - The euthyroid sick syndrome is reported to exist in acute myocardial infarction(AMI). Previous reports showed serum levels of triiodothyronine(T3) are low and thyroid stimulating hormone(TSH) is normal or subnormal levels in patients with AMI. However, the mechanism of altered thyroid hormone metabolism is unknown. Interleukin-6(IL-6) is reported to be a key role in the pathogenesis of AMI and euthyroid sick syndrome. We measured circulating TSH, free T3(FT3), free thyroxine (FT4), IL-6, soluble IL-6 receptor, soluble transducing 130-kD glycoprotein, atrial natriuretic peptide(ANP) and brain natriuretic peptide in 25 patients and 32 healthy subjects. Circulating FT3 levels in patients with AMI became lower than in control group(p < 0.05). IL-6 levels were significantly(p < 0.05) higher than those of healthy subjects. The peak levels of IL-6 was 30.5 +/- 46.9 pg/ml at 25-27 hours(the first peak) and 64.4 +/- 24.6 pg/ml at 70-72 hours(the second peak). FT3 was negatively related to IL-6(p < 0.05) and hANP(p < 0.05) in patients with AMI. These results indicate that the lower levels of FT3 show the greater severity of AMI. We conclude that euthyroid sick syndrome occurs in patients with AMI and euthyroid sick syndrome may regulated by IL-6 through suppressed of thyroid function. PMID- 10804838 TI - [Quantitation of human immunodeficiency virus type 1 RNA by ultrasensitive method in the analysis of samples with low viral load]. AB - With the introduction of combination therapy to the patients with Human Immunodeficiency Virus Type 1(HIV-1) infection, HIV-1 RNA levels in plasma have been reduced, frequently to less than limit of quantitation(400 copies/ml). To achieve enhanced sensitivity, a modified specimen preparation procedure that allows the quantitation of plasma HIV-1 RNA levels as low as 50 copies/ml was developed. Of 67 samples with less than 400 copies/ml of HIV-1 RNA by standard method, 39(58.2%) were not detected by the "ultrasensitive" method. Among 5 samples obtained from patients who were treated with 2 nucleoside reverse transcriptase inhibitors, 4 samples still had detectable copies of HIV-1 RNA. Those suggest that the ultrasensitive assay for HIV-1 RNA has advantage for clinical practice. PMID- 10804839 TI - [Seroprevalence of maedi-visna and border disease in Switzerland]. AB - 3866 sheep from 226 flocks of breeding associations and 1218 sheep from 15 independent sheep owners were tested for the presence of serum antibodies against Maedi-Visna and Border Disease viruses. The flocks were randomly selected based on the relative proportion and the geographical distribution of the 4 predominant Swiss sheep breeds (Braunkopfiges Fleischschaf, Schwarzbraunes Berg- und Juraschaf, Walliser Schwarznasenschaf, Weisses Alpenschaf). Additionally two smaller breeds were included in the study (Charollais Suisse, Milchschafe). Sera of all sheep older than 1 year were collected together with data characterizing host and management factors. The sera were tested using established ELISAs for detection of antibodies to Maedi-Visna and Bovine Virus Diarrhea/Border Disease viruses. ELISA results of Maedi-Visna serology were confirmed by immunoblotting. 9% of the sheep of breeding associations were antibody-positive for Maedi-Visna virus. The results of the different breeds varied between 0.4% and 36%. A multiple logistic regression procedure identified breed, age, airing in barns, herd size, pasturing on alps and way of keeping the animals during winter as associated factors with individual serostatus. The prevalence of antibodies to Border Disease was 20% in sheep of breeding associations and 65% in those of independent sheep owners. PMID- 10804840 TI - [Pilot project for eradicating maedi-visna in Walliser blacknose sheep]. AB - Maedi-Visna is a lentiviral disease of sheep with a worldwide distribution. The transmission of the virus occurs primarily via colostrum and milk from the infected ewe to its newborn lamb but also horizontally between sheep. The most obvious clinical symptoms are progressive dyspnea and emaciation. In this prospective study an eradication based on serological testing and removing of seropositive animals was performed in 24 flocks of sheep of the breed "Walliser Schwarznasenschafe" leading to a reduction of the seroprevalence from 36% to 1% within two years. The control group consisted of 21 flocks of sheep. Lambs of seropositive ewes had a 7.6 times higher risk to seroconvert within their first two years of life compared to those of seronegative ewes. The dynamics of the spread of the infection were studied in birth cohort groups. Cohort animals of seropositive ewes showed an obvious trend to seroconvert slowly. Seropositive ewes had a significantly lower reproduction rate and their lambs suffered from significantly higher death and lower growth rates, probably due to a reduced milk production, resulting in economic losses. PMID- 10804841 TI - [The role of common starlings (Sturnus vulgaris) in the epidemiology of bacterial, potentially human pathogenic, disease agents]. AB - Since a long time a public garden in Basel is known as a site for overnight accommodation and assembly of starlings. The birds cause an immense faecal contamination of the park and the neighbouring district. A nursery and a primary school are directly affected. To evaluate the health risk coming from the starlings droppings for the population, particularly for the children and to assess the role of starlings in the transmission of diseases to humans and in the epidemiology of human diseases the presence of human bacterial pathogens in the faeces of starlings was determined. Some of the isolated strains were further typed and compared to strains of human origin. C. jejuni, L. monocytogenes and C. psittaci were most often found. The typing of some C. jejuni and L. monocytogenes isolates showed a great variety of geno-, sero- respectively phage types that did not belong to the strains most often found in isolates of human origin. Starlings can harbour human pathogens and therefore a potential risk of infection comes from their droppings. It seems however rather improbable, that these birds present a constant direct source of infection for human beings. PMID- 10804842 TI - Determination of Lufenuron in canine skin layers by radioluminography. AB - In a bioavailability study in dogs we investigated the quantity of [14C] Lufenuron and its metabolites in the skin by radioluminography. Two Beagle dogs were orally administered 10 mg [14C]-Lufenuron per kg body weight. 21 days later they were euthanized. Skin specimens were taken at 6 different sites. Sections of these skin specimens were exposed on a phosphorus imaging plate and the radiolabeled Lufenuron content visualized. Radioluminographical analysis showed that [14C]-Lufenuron appeared in the subcutaneous layer of the skin. We did not discover any activity on the skin surface. PMID- 10804843 TI - [Evidence-based medicine in routine clinical practice]. PMID- 10804844 TI - [Evidence-based medicine: an approach without any weakness?]. AB - Evidence-based medicine is a methodological approach giving access to the best information derived from clinical research for an individual patient. It requires the formulation of a question, a strategy to search for the best information, the selection of the latter, its critical appraisal and its application to the patient. The qualities, but also the limitations of this approach are discussed. PMID- 10804845 TI - [Medical treatment of low back pain according to evidence-based medicine]. AB - Results of therapy of low back pain with drugs (NSAIDs), locally injected corticosteroids, active physical therapy, back school and chiropractic are assessed according to "Evidence Based Medicine". The assessment after a trial period of therapy is difficult because clinical symptoms and signs of low back pain are non specific. There is no good correlation of clinical signs with the radiological images. The therapy is empiric since controlled randomized clinical studies have until now not shown that drugs, corticosteroids (epidural or intra facet-joint injections), back school, chiropractic or physical therapy give results better than with placebo. PMID- 10804846 TI - [Evidence-based medicine and arterial hypertension: reasonable or unreasonable marriage?]. AB - Rational medical decisions should be based on the best possible evidence. Yet clinical trial results may not reflect conditions in actual practice. In hypertension, for example, the evidence-based approach to medical care involves the explicit use of evidence on the magnitude of the effects of interventions to influence treatment decisions. Evidence-based medicine is a paradigm challenging our medical practice, as foundation it uses proofs originating from scientific research and controlled randomized trials, more than intuition, clinical experience and pathophysiologic rationale. In this article, we analyze two clinical problems in hypertension generating current questions. We try to use this method in order to obtain pertinent answers for a given patient. We observe that the application of EBM has some limitations. Very restrictive inclusion criteria in some studies make the sample of patients studied not comparable with the individuals in front of us. On the other hand, for other sub-groups of patients, results obtained from EBM are very useful and give valuable information. For a good application of this paradigm critical reading of literature is essential, in order to balance its importance. PMID- 10804847 TI - [Nebivolol: a beta blocker with vasodilator properties]. AB - Nebivolol is a new cardioselective beta-blocking agent possessing vasodilatory properties involving the endothelium. This compound is a dl-racemic mixture. The d-enantiomer is responsible for the beta-blocking properties whereas the l enantiomer induces a vasodilation via a nitric oxide (NO) mechanism. Nebivolol is an unique agent that appears promising for the management of patients with hypertension, coronary heart disease or congestive heart failure. PMID- 10804848 TI - Strategies of bacterial interaction with eukaryotic cells. PMID- 10804849 TI - Microtubule dependent invasion pathways of bacteria. PMID- 10804850 TI - Interaction of enteropathogenic Escherichia coli with host cells. PMID- 10804851 TI - E. coli invasion of brain microvascular endothelial cells as a pathogenetic basis of meningitis. AB - A major limitation to advances in prevention and therapy of bacterial meningitis is our incomplete understanding of the pathogenesis of this disease. Successful isolation and cultivation of BMEC, which constitute the blood brain barrier, and the development of experimental hematogenous meningitis animal model, which mimics closely the pathogenesis of human meningitis, enabled us to dissect the pathogenetic mechanisms of bacterial meningitis. We have shown for the first time using E. coli as a paradigm the mechanisms of bacterial crossing of the blood brain barrier into the central nervous system. We have shown that invasion of BMEC is a requirement for E. coli K1 crossing of the blood-brain barrier in vivo (Prasadarao et al., 1996b; Huang et al., 1995). We have identified several novel E. coli proteins (i.e., Ibe10, Ibe7, and Ibe23) contributing to invasion of BMEC. We have also established a novel phenotype, i.e., invasion of BMEC, of a well known major E. coli protein, OmpA. In addition, we have shown that some of these E. coli proteins (i.e., OmpA, Ibe10) interact with novel endothelial receptors present on BMEC, not on systemic vascular endothelial cells. Further understanding and characterization of these E. coli-BMEC interactions should allow us to develop novel strategies to prevent this serious infection. In addition, the in vitro and in vivo models of the blood-brain barrier and the information derived from our study should be beneficial to investigating the pathogenesis of meningitis due to other organisms such as group B streptococci, Listeria monocytogenes, Streptococcus pneumoniae and Citrobacter. PMID- 10804852 TI - Host cell invasion by pathogenic Neisseriae. AB - As outlined in this review, various experimental techniques have been employed in an attempt to understand neisserial pathogenesis. In vitro genetic analysis has been used to study the genetic basis for the structural variability of cell surface components. Transformed or primary epithelial cell cultures have provided the simplest model to analyze bacterial adherence and invasion, while the infection of polarized epithelial monolayers, fallopian tube and nasopharyngeal organ cultures, and ureteral tissue have each been used to more closely represent the events which occur in vivo. Finally, the in vivo infection of human volunteers with N. gonorrhoeae has provided a powerful means to confirm and expand the results obtained in vitro. By these various approaches, a number of neisserial adhesins (i.e. pilli, Opa, Opc and P36) and additional putative virulence determinants which affect bacterial adherence and invasion into host cells (i.e. LOS, capsule, PorB) have been identified. Clearly, neisserial surface variation serves as an adaptive mechanism which can modulate tissue tropism, immune evasion and survival in the changing host environment. Important progress has been made in recent years with respect to the host cellular receptors and subsequent signal transduction processes which are involved in neisserial adherence, invasion and transcytosis. This has led to the identification of (i) CD46 as a receptor for pilus which allows adherence to epithelial and endothelial cells, (ii) HSPGs, in cooperation with vitronectin and fibronectin, as receptors for a particular subset of Opa proteins and Opc, which may both mediate invasion into most epithelial and endothelial cells, and (iii) CD66 as the receptors for most Opa variants, potentially being involved in cellular interactions including adherence, invasion and transcytosis with epithelial, endothelial and phagocytic cells. As most of these data have been obtained using transformed cell lines growing in vitro, attempts must be made to translate these basic observations into a more natural situation. It can be expected that the successful ongoing integration of laboratory findings from the various infection models with human volunteer studies will further increase our understanding of the biology of neisserial infection. Perhaps the most difficult but also most rewarding challenge for the future will be to use volunteer studies to identify and understand the role of host factors which are important for the infectious process. Hopefully, insights gained from each of these studies will reveal new and useful strategies for the preventive and/or therapeutic intervention into infection and disease by these fascinating microbes. PMID- 10804854 TI - Host cell invasion by Streptococcus pneumoniae. PMID- 10804853 TI - Bartonella interactions with host cells. PMID- 10804855 TI - High frequency invasion of mammalian cells by beta hemolytic streptococci. PMID- 10804856 TI - Nocardia asteroides as an invasive, intracellular pathogen of the brain and lungs. PMID- 10804857 TI - Mycoplasma interaction with eukaryotic cells. PMID- 10804858 TI - Mycobacterial invasion of epithelial cells. PMID- 10804859 TI - Invasion of epithelial cells by bacterial pathogens the paradigm of Shigella. PMID- 10804860 TI - Salmonella invasion of non-phagocytic cells. PMID- 10804861 TI - Salmonella interactions with professional phagocytes. PMID- 10804862 TI - Interaction of Yersinia with host cells. PMID- 10804863 TI - Invasion of mammalian and protozoan cells by Legionella pneumophila. PMID- 10804864 TI - Internalization of Listeria monocytogenes by nonprofessional and professional phagocytes. PMID- 10804865 TI - Host-plant invasion by rhizobia. PMID- 10804866 TI - Chlamydia internalization and intracellular fate. PMID- 10804867 TI - Interaction of rickettsiae with eukaryotic cells. Adhesion, entry, intracellular growth, and host cell responses. PMID- 10804868 TI - Ehrlichial strategy for survival and proliferation in leukocytes. PMID- 10804869 TI - DNA vaccine delivery by attenuated intracellular bacteria. AB - Vaccination by intramuscular or intradermal injection of antigen-encoding DNA is a promising new approach leading to strong cellular and humoral immune responses. Since bone-marrow derived antigen presenting cells (APC) seem to induce these immune responses after migration to the spleen, it is desirable to deliver DNA vaccines directly to splenic APC. Recently, attenuated intracellular bacteria have been exploited for the introduction of DNA vaccine vectors into different cell types in vitro as well as in vivo and offer an attractive alternative to the direct inoculation of naked plasmid DNA. PMID- 10804870 TI - Vaccines against intracellular pathogens. PMID- 10804871 TI - Identification and in situ detection of intracellular bacteria in the environment. PMID- 10804872 TI - New approaches for diagnosis of infections by intracellular bacteria. PMID- 10804873 TI - [Genetic predisposition for alcoholism]. AB - A number of socio-economic, cultural, biobehavioral factors and ethnic/gender differences are among the strongest determinants of drinking patterns in a society. Both epidemiological and clinical studies have implicated the excessive use of alcohol in the risk of developing a variety of organ, neuronal and metabolic disorders. Alcohol abuse related metabolic derangements affect almost all body organs and their functions. Race and gender differences in drinking patterns may play an important role in the development of medical conditions associated with alcohol abuse. The incidence of alcoholism in a community is influenced by per capita alcohol consumption and covariates with the relative price and availability of alcoholic drinks. The majority of the family, twin and adoption studies suggest that alcoholism is familial, a significant proportion of which can be attributed to genetic factors. The question is how much of the variance is explained by genetic factors and to what degree is this genetically mediated disorder moderated by personal characteristics. Among the most salient personal characteristics moderating, the genetic vulnerability may be factors such as age, ethnicity, and presence of psychiatric co morbidity. Cultural factors and familial environmental factors are most likely predictors as well. PMID- 10804874 TI - [Markers for excessive alcohol use (screening)]. AB - It is well-known that early diagnosis in addiction leads to a better outcome and prevents psychosocial and medical illness and disability as well as costs. It would be important to have a gold standard for the diagnosis for alcoholism because of the consequences of this diagnosis for both the patient and the physician. In the last 15 years there were world-wide efforts to find biological markers for alcoholism and alcohol abuse. The results, however, were rather poor. With the exception of the relatively new and expensive CDT TEST (Carbohydrate deficient transferrin) and some changes in established questionnaires (shortenings) we have used the same screening tests for decades. The relationship between the patient and the physician, a detailed medical history and experience of the physician cannot be replaced by tests. The Plinius Major Society recommends in its Guidelines the CAGE questionnaire. In medical settings and in primary care the MALT or AUDIT are more informative. As laboratory markers the Plinius Major Society still recommends: gamma-GT, MCV, GOT/GPT (ASAT/ALAT) and CDT. These tests are only useful if normal values of the particular laboratory are given. PMID- 10804875 TI - [Alcohol and the brain--morphological and functional brain changes]. AB - It has been established long ago through autopsy studies that chronic alcohol consumption may lead to severe organic brain damage in about 10% of the patients. The recent rapid development of novel tools for structural and functional brain imaging has enabled researchers to expand upon this early knowledge. Some of these newer studies have demonstrated that chronic alcohol consumption causes cortical volume loss, which is partly reversible with abstinence. Less known but of utmost clinical relevance are non-organic neurocognitive dysfunctions, which have only been described recently. Functional studies revealed that alcohol may lead to patterns of reduced activity in brain activation which correlate with the atrophy and take months to years to recover. PMID- 10804876 TI - [Alcohol and the peripheral nervous system]. AB - The most frequent consequence of chronic alcohol intake is a toxic polyneuropathy. It results from inadequate nutrition, mainly deficiency of thiamine and other B vitamins. Additionally there is a direct neurotoxic effect of ethanol. Signs and symptoms are 1. distal sensory disturbances with pain, paresthesia, and numbness in a glove and stockings-pattern, 2. weakness and atrophy of distal muscles, pronounced in the lower limbs, 3. loss of tendon jerks, 4. affection of autonomic fibers. Therapy consists in absolute alcohol abstinence, high-caloric nutrition, parenteral thiamine and other vitamins. Against paresthesia and pain, carbamazepine, salicylates, amitryptiline are effective. Parenteral tioctacid may be tried. The prognosis of alcoholic polyneuropathy is favorable, with alcohol abstinence, within several months up to a few years. In chronic alcoholic patients peripheral nerves frequently are injured by compression during alcohol intoxication. Peroneal nerve lesions result from compression in the region of the neck of the fibula during a prolonged lying position, the radial nerve is injured during sitting with the upper arm placed on the backrest of a bench. Usually pressure palsies resolve spontaneously. Rhabdomyolysis is a rare but life-threatening complication of alcoholic delirium. Symptoms are severe muscle pain, swelling of extremities, pigmenturia. The major complications of rhabdomyolysis are renal and respiratory failure, and cardiac arrhythmias due to electrolyte imbalance. Intensive care is needed with control of hyperkalemia, hydration, alkalinization of urine, hemodialysis if indicated. PMID- 10804877 TI - [Alcohol, the heart and cardiovascular system]. AB - Any advice about the consumption of alcohol must take into account not only the relation between alcohol and cardiovascular disease but also the well-known association of heavy consumption of alcohol with a large number of health risks. Numerous observational studies have consistently demonstrated a reduction of coronary heart disease with moderate consumption of alcohol. Consumption of 1 or 2 drinks per day is associated with a reduction in risk of dying from coronary heart disease of approx. 30-50%. A drink equivalent amounts to a 12-ounce bottle of beer, a 4-ounce glass of wine, and a 1.5-ounce shot of 80-proof spirits. It has been repeatedly demonstrated that there is a J-shaped relation between alcohol consumption and total mortality. The lowest mortality occurs in those who consume 1 or 2 drinks per day. A stepwise decline in coronary heart disease death occurs with increasing drinks per day. Because coronary heart disease accounts for 1/3 or more of total death, people with no alcohol consumption have higher total mortality than those drinking 1 to 2 drinks per day. Conversely, mortality due to a large number of other diseases increases with an increasing number of drinks consumed per day. The protective effects of alcoholic against coronary heart disease are mainly related to an increase in HDL cholesterol. A number of other mechanisms have been proposed including effects of alcohol on blood clotting and non-alcoholic components of alcoholic beverages, particularly in red wine and dark beer, which may have antioxidant properties. Harmful effects of alcohol on the cardiovascular system include congestive cardiomyopathy, systemic hypertension and cerebral vascular incidents. There is a direct correlation between the amount of alcohol consumed during lifetime and a reduction in left ventricular ejection fraction. PMID- 10804878 TI - [Alcohol, lipid metabolism and body weight]. AB - Regular alcohol consumption has multiple effects on lipid metabolism and basically all lipid fractions in the blood are affected by alcohol. The effects depend on the dose and the regularity of intake as well as the liver function. Moderate amounts of alcohol lead to an increase in HDL-cholesterol as well as in some populations to a lowering in the Lp(a) concentration. High consumption levels lead to a decline in most lipoprotein fractions due to hepatopathy. Due to the suppression of lipid oxidation moderate alcohol consumption has to be regarded as a risk factor for obesity and weight gain. The non-oxidized lipid calories are deposited preferentially in the abdominal area. These effects can be compensated by a reduction of fat intake in a ratio of at least 1:1 to the amount of the ingested calories. PMID- 10804879 TI - [Alcohol, the gastrointestinal tract and pancreas]. AB - The intake of larger quantities of alcoholic beverages leads to manifold functional disturbances and organ injury in the upper gastrointestinal tract. These damaging effects of alcohol are frequently the cause of complaints, such as heart burn, symptoms of dyspepsia and diarrhoea. Examples of more pronounced organ injury which can occur even following a single episode of heavy drinking are tears in the mucosa at the junction of the esophagus and the stomach (Mallory Weiss-lesion) and hemorrhagic erosions in the stomach and/or the duodenum which may lead to massive bleeding. In the small intestine alcohol abuse interferes with the absorption of glucose, amino acids, lipids, water, sodium and vitamins (especially thiamine and folic acid). This inhibition of absorption of nutrients may contribute to nutritional deficiencies frequently observed in alcoholics. Acute alcohol ingestion can also damage the mucosa in the upper region of the small intestine and may lead to the disruption of the tips of the villi. Chronic alcohol abuse increases markedly the prevalence of bacterial overgrowth in the small intestine. The findings of human and animal studies suggest that the mucosal injury together with bacterial overgrowth favour the following sequence of events: Alcohol induced mucosal injury in the small intestine increases the permeability of the mucosa to macromolecules, such as endotoxin and/or other bacterial toxins, into the blood or lymph. This results in the release of potentially toxic cytokines and other mediators like Kupfer cells and other phagocytes. These cytokines and other mediators, in turn, exert multiple injurious effects on the microcirculation and membranes. The result is cell damage and even cell death (apoptosis, necrosis) in the liver and other organs. Chronic alcohol abuse is one of the most important risk factors for the development of cancers of the tongue, larynx, pharynx and esophagus. In many countries alcohol abuse is the most important cause for the development of chronic pancreatitis. In the initial phase the disease is frequently characterised by episodes of 'acute' pancreatitis. These episodes develop only on the basis of prolonged alcohol abuse leading to subclinical damage of the gland. The latter is found in about 20-50% of patients with chronic alcohol abuse while the clinically overt pancreatitis is observed in only 1%-3% of alcoholics. Despite numerous studies performed in animal experiments and man the pathogenesis of alcoholic pancreatitis until now has not been clarified. PMID- 10804880 TI - [Alcohol and drug interactions]. AB - Alcohol metabolism occurs mainly in the liver, where in abstainers the alcoholdehydrogenase (ADH) pathway plays the major role. After chronic alcohol consumption, the microsomal ethanol-oxidizing system (MEOS), involving the ethanol-inducible cytochrome P450 2E1, increases in importance with a four- to ten-fold increase in the contribution to alcohol metabolism. Because of the fact that this enzyme system catalyses not only the metabolism of ethanol but also activates a great number of drugs, it is a very common site of alcohol-drug interactions. Clinically relevant interactions will be discussed. Only a small amount of alcohol is metabolized outside the liver, mainly in the stomach by gastric ADH, which leads to the so-called first-pass metabolism of ethanol. Its significance in alcohol metabolism is reviewed. The only way to prevent severe alcohol-drug interactions is to make medical doctors as well as their patients more aware of these possible secondary effects. PMID- 10804881 TI - [Alcohol and the liver]. AB - Since ethanol metabolism predominantly takes place in the liver it is not surprising that hepatic intermediary metabolism is strikingly influenced. Alcohol is metabolized via three enzyme systems: alcohol dehydrogenase (ADH), microsome ethanol oxidizing system (MEOS) and catalase. The ADH reaction produces reducing equivalents as NADH which results in various metabolic disorders such as hyperproteinemia IV and V, hypoglycaemia, lactacidosis, hyperuricaemia, and certain forms of porphyria. The metabolism of hormones is also disturbed. Alcohol fatty liver is a direct consequence of NADH production. Alcoholic liver disease comprises of fatty liver, alcoholic hepatitis and cirrhosis. Risk factors of alcoholic liver disease are the amount of alcohol consumed, drinking pattern, female gender and certain genetic predispositions. Alcoholic hepatitis is characterized by a typical clinical and laboratory feature, and specific heaptic morphology. Poor prognostic factors are continuous alcohol consumption, cholestatis and perivenular fibrosis. Alcoholic cirrhosis has similar complications as cirrhosis of other etiology. Therapy includes abstinence, antioxidative drugs, steroids, and S-adenosylmethionine. Liver transplantation is of long-term benefit. PMID- 10804882 TI - [Skin manifestations of alcoholic liver damage]. AB - Alcohol-induced diseases of the liver, such as fatty liver, hepatitis and cirrhosis with the potential development of hepato-cellular carcinoma can cause many effects on the skin. Even though they are not caused by excessive alcohol alone, but also by other diseases of the liver or other diseases of internal organs, an experienced person will be able to carry out specific diagnostic procedures. Skin symptoms due to liver diseases include 1. Vascular changes, such as spider nevi, teleangiectasias and palmar erythema. 2. Nail changes, particularly white nails. 3. Changes of the mucous membranes, i.e. glossy tongue. 4. Changes due to altered hormones, particularly gyneco-mastia, female distribution of hair and testicular atrophy and 5. Changes in the color of the skin like icterus and melanosis cutis. Rarely pruritus and other diseases of the skin are seen, such as porphyria cutanea tarda, which is often caused by an altered liver function. In the final stages of alcoholism, the neglect of personal hygiene particularly of the skin is evident (cutis vagantium). Since the exact mechanism of the skin symptoms remains obscure, it is difficult to evaluate the significance. Most often they do not correlate with the severity of the liver disease. PMID- 10804883 TI - [Alcohol and cancer]. AB - Numerous studies have identified alcohol as the main risk factor for cancer of the upper GI tract such as pharynx, larynx and esophagus. Alcohol consumption is also associated with an enhanced risk for hepatocellular, breast and colorectal cancer. Animal experiments have demonstrated, that alcohol per se is not carcinogenic but rather exerts tumor-promoting and cocarcinogenic properties in animal models. These findings may be explained by recent experimental and epidemiological studies which have focused on the first metabolite acetaldehyde as the main tumorigenic substance of alcohol abuse. Other possible tumor promoting effects of alcohol are the existence of carcinogenic congeners in alcoholic beverages, an increased solubility of carcinogens by alcohol, a reduced intake and bioavailability of possible cancer protective substances, an inhibition of detoxification of carcinogenic compounds or increased activation of procarcinogens by cytochrome P450IIE1 which is induced by alcohol and a suppressed immune function. Alcohol also leads to a direct local mucosal damage, which results in a chronically increased cell division and thus an increased cancer risk. PMID- 10804884 TI - [Alcohol and sleep disorders]. AB - Despite an initial sedative effect, alcohol disrupts sleep persistently and should not be used as a sleeping aid. Nocturnal withdrawal symptoms may lead to an increased duration of wakefulness, and to tachycardia and sweating in the second half of the night. It is not known by which mechanism alcohol affects sleep; however, effects do not appear to depend on the stimulation of benzodiazepine receptors or the antagonism at adenosine receptors. Alcohol can exacerbate primary sleep disturbances such as sleep apnea and nocturnal myoclonus, and thereby contribute to excessive daytime sleepiness. The sleep of alcoholic patients is characterized by increased sleep latency, and reduced sleep efficiency, total sleep time, slow wave sleep and non-REM sleep. Even during abstinence, the changes in sleep architecture can persist for months or years, and might contribute to a relapse into alcoholism. The use of benzodiazepines or other hypnotics to treat alcohol-related sleep disturbances is not recommended. PMID- 10804885 TI - [Alcohol and pregnancy--embryopathy and alcohol effects]. AB - Fetal alcohol syndrome (FAS) is a specific polydystrophic pattern of malformations with the following diagnostic criteria: 1. Maternal alcohol dependence or alcohol abuse during pregnancy. 2. Pre- and postnatal deficiency of growth in weight, height and head circumference. 3. Multiple minor and major anomalies recognizable mainly at a typical face. 4. Structural injuries and changes at the central nervous system with complex brain dysfunction combining elements of cognitive impairments, behavioral disturbance and neurological damage. Fetal alcohol effects (FAE) or so-called "alcohol-related neurodevelopmental disorders" (ARND) with predominant neurotoxic effects and a large spectrum of cerebral dysfunctions are manifold more frequent than the full blown FAS. These remain mostly unrecognized, overlooked and they are difficult to be diagnosed, the symptoms being unspecific. Alcohol in pregnancy is nowadays the most important and the most frequent toxic substance for the embryo and the fetus and one of the most frequent causes of mental retardation. The longlasting and irreversible consequences refer to school development, social maturation, social behaviour and later life-style. The diagnosis is based on the careful maternal history and on the clinical findings; there are no biochemical parameters of assessment. The risk of addiction development in these children is assumed to be more than 20 percent. PMID- 10804886 TI - [Treatment of alcoholism]. AB - In view of the many non-specific symptoms a diagnosis of alcoholism can only be made by means of a detailed history and a medical checkup. Simple, standardized questions may often be sufficient. After a diagnosis a consultation with a family physician and drugs, such as disulfiram, acamprosate and naltrexone have been proved to be a highly efficient therapy. Therefore, it is worthwhile to treat alcoholics in the general practice setting. PMID- 10804887 TI - [Alcohol withdrawal syndrome and its treatment]. AB - When alcoholics decrease or interrupt alcohol intake abruptly, they will in general experience alcohol withdrawal symptoms. Clinically in most cases it develops a vegetative syndrome with gastroenteropathy, cardiovascular diseases, neurological and psychopathological symptom. Usually alcohol withdrawal symptoms abate after four to seven days, longer courses are rare. Application of drugs is required in approximately one third to one half of the patients. A variety of drugs was suggested for the treatment of alcohol withdrawal. In the first hours of alcohol detoxification, the sensitivity of epinephrine receptors is reduced, but rises afterwards substantially. The number of NMDA-receptors increases during chronic intoxication with ethanol. The standard therapy in Europe (except of Great Britain) is an oral mono therapy with clomethiazole in a dose which depends on the severity of the symptoms. Severe withdrawal symptoms may require treatment on an intensive care unit with infusion therapy, e.g. in the context of a delirium tremens, which represents a life-threatening status. In this case, benzodiazepines have been used successfully as an alternative to clomethiazole. PMID- 10804888 TI - [Anesthesiologic aspects of chronic alcohol abuse]. AB - The prevalence of alcoholism exceeds 20% in surgical disciplines. Chronic alcoholics are endangered by an increased postoperative morbidity and mortality. Due to an elaborated preoperative alcoholism-related diagnostic evaluation (alcoholism-related questionnaire and laboratory markers) and an adequate perioperative management (prophylactic treatment of alcohol withdrawal symptoms and symptom-orientated therapy of infections, cardiac complications and bleeding disorders), intercurrent complications may be reduced and postoperative outcome improved. PMID- 10804889 TI - [Alcohol--forensic aspects]. AB - Alcohol (resp. ethanol) in forensic medicine includes mainly the following areas and fields of problems: analysis (in particular blood- and breath alcohol), ability to drive resp. DUI/DWI, impairment of responsibility, alcohol use, abuse and dependence, intoxication, alcohol as direct and indirect cause of death and organ damage. A short overview presents these problem areas of alcohol use and misuse as well as results of alcohol research ("ethanology") which may be relevant for clinical use. PMID- 10804890 TI - [Alcohol at the work site]. AB - In Switzerland in large companies alcohol prevention programmes are wide spread. Their basic aim is to reduce hidden costs and to improve security at the workplace. To reach these goals early detection of employees with alcohol problems has been introduced and referral systems to therapeutic measures have been developed. Many of the alcohol prevention programmes, however, do not meet the standards of good practice discussed in this article. In addition, the cost efficiency of these programmes could rarely be proved. A noticeable lack of alcohol prevention programmes exists, however, in small and medium companies. PMID- 10804891 TI - A classroom assignment. PMID- 10804892 TI - A typology of residential fire survivors' multidimensional needs. AB - This article presents an analysis of the tangible, psychological, and general needs of adults and their children reported by residential fire survivors approximately 14 weeks postfire. Three hundred and seven survivors, who identified that they needed help for themselves or their children, differed from 133 survivors who did not request help. Those needing help were more likely to be women with children younger than age 18 living in their household, have low income status, less education, and to have already received services from church groups. The classification of self-identified needs of fire survivors included the need for specific tangible and social service assistance, psychological and spiritual support, and nonspecific assistance. This classification contributes to our understanding of the relationship between needs and loss, grief, and changes in family and life situations postfire. PMID- 10804893 TI - Self-disclosure of HPV infection to sexual partners. AB - Disclosure decisions of persons with genital HPV infection were examined. The research questions focused on relationships among knowledge of transmission, beliefs about the obligation to disclose knowledge of HPV infection to sexual partners, factors that influence the disclosure decision, and evaluations of the decision. Participants were 92 persons diagnosed with HPV 6 months prior to data collection. Sixty-three women and 29 men whose average age was 23.1 years constituted the sample. A subset of 48 persons provided detailed information about their reasons for disclosing or not disclosing knowledge of HPV and subsequent evaluation of their behavior. Knowledge of HPV transmission was adequate; however, there was no relationship between transmission knowledge and disclosure beliefs. Participants tended to disclose the presence of HPV to partners at point of diagnosis, but not to new partners 6 months later. Both disclosers and non-+disclosers felt positively about their decision. Possible changes in clinical interventions for persons with HPV infection are discussed. PMID- 10804894 TI - The meaning of hope in healthy, non-religious Swedes. AB - This study aims to illuminate the meaning of the lived experience of hope as narrated by healthy Swedish adults, who considered themselves as non-religious. Twenty-four interviewees were asked to narrate their experience of hope. A phenomenological-hermeneutical analysis was performed and the meaning of the lived experience of hope was interpreted as an internal process (i.e., hope related to being), which is linked to the experience of meaning and the awareness of one's possibilities in life. This awareness releases energy and activates thoughts and feelings, enabling the person to make good and meaningful choices. The process generates a sense of independence, freedom, and inner peace. Hope related to being is a prerequisite for an external process (i.e., setting goals). Setting goals comprises action and the expectation of a positive outcome. Hope related to doing nurtures hope related to being. The findings also revealed that hope related to doing is reconstructed throughout the life process. PMID- 10804895 TI - Feeding and non-feeding interactions of mothers and prematures. AB - The interactions between mothers and premature infants during feeding and nonfeeding periods were explored. Twenty-nine premature infants and their mothers were observed interacting for 1 hour in their homes at 6 months corrected for prematurity. Mothers were more likely to engage in the following behaviors involving close contact during feeding: looking at the infant, holding the infant, having body contact, and rocking their infants. They interacted with their infants 96% of feeding time. During nonfeeding periods, they were more likely to engage in more distal behaviors (e.g., gesturing, touching, and playing with the infant) and spent only 67% of the time interacting with the infant. Infants were more likely to be alert, vocalize, play with objects, express negative affect, and locomote during nonfeeding, and they were more likely to be drowsy or asleep during feeding. Therefore, a complete understanding of interactions between mothers and prematures can only come from examining both feeding and nonfeeding periods. PMID- 10804896 TI - Do pressure relief cushions really relieve pressure? AB - In a laboratory setting, interface pressures of 29 cushions and a sheepskin were measured on 20 healthy volunteers. Each participant was seated in an upright posture with their back against the back of the chair, hands resting on the lap, knees bent at an angle of 90 degrees, and feet resting on the floor. Only 13 cushions had any pressure-reducing effect. Gel cushions and sheepskins appear to have no pressure-reducing effect. The category of foam includes both cushions that reduce interface pressure very well and cushions that increase interface pressure. The lowest interface pressures were measured on air cushions and on some foam cushions. PMID- 10804897 TI - The concept of theme as used in qualitative nursing research. AB - The concept of theme is critical to the accurate interpretation of qualitative data. A literature review of qualitative research methodology and nursing research studies reveals considerable diversity in the identification of themes, the interpretation of the concept, and its function in data analysis. Part of the problem is the transfer of research methods from other disciplines to the study of nursing phenomena. The transfer often results in a blending of distinct research methods that compromises methodological rigor in data analysis and theory generation. A definition of theme is developed from the literature review and interdisciplinary definitions and descriptions. Five aspects of a theme and criteria foundational to the definition and concept of theme are identified. Implications of the concept of theme are presented for the conduct and application of research findings to the practice and development of nursing science. PMID- 10804899 TI - Y2K--a colossal hoax? PMID- 10804898 TI - Supraesophageal complications of gastroesophageal reflux. AB - Supraesophageal complications of GERD have become more commonly recognized or suspected by physicians. However, the direct association between these complications and GERD has often been difficult, if not impossible, to establish. Furthermore, the majority of patients with suspected supraesophageal complications of GERD do not have either the characteristic symptoms of heartburn and regurgitation or the definitive findings of esophageal inflammation, which would help reinforce the suspicion of a connection between the supraesophageal complications and GERD. Frequent acid reflux has been shown in patients with various bron-chopulmonary, laryngopharyngeal, or oral cavity disorders. GERD is one of the most common gastrointestinal complaints in the population. It is possible that the supraesophageal problems and acid reflux are mutually independent disorders that occur in the same person. The suspected mechanisms of GERD-related supraesophageal complications appear to be directed through two pathways: by a vagal reflex between the esophagus and tracheobronchial tree triggered by acid reflux or by microaspiration that causes contact damage to mucosal surfaces. The most useful diagnostic modality available to the clinician to aid in the diagnosis of supraesophageal GERD complications is the ambulatory pH recording technique. However, the sensitivity and specificity of this test for recording esophageal or pharyngeal acid reflux events has been critically challenged. Despite the many clinical studies that support the theory that GER has a role in suspected supraesophageal complications, only 1 long-term prospective controlled study of a large group of patients with asthma has shown the positive effects of the elimination of acid reflux. With the focus now on "outcomes medicine," there is a serious need for appropriately designed, controlled studies to answer the many questions surrounding a cause-and-effect association between acid reflux and supraesophageal disorders. Because of the lack of convincing proof between acid reflux and suspected supraesophageal complications, the physician must resort to an intent-to-treat strategy as both a primary therapy and a diagnostic trial. High-dose PPI therapy for prolonged periods is the recognized conservative therapy. Operative therapy (i.e., fundoplication operation) is the procedure of choice when overt regurgitation occurs or when medical therapy, although successful, is not practical for long periods. Controlled, well-designed clinical trials and more sophisticated techniques to measure and quantify acid reflux are crucial in the future to help determine which patients with suspected supraesophageal complications actually have acid reflux as a primary cause. The medical community needs to be alerted to the possibility of an association between GERD and supra-esophageal complications so that patients with a GERD-related complication will be recognized and effectively treated. PMID- 10804900 TI - The catastrophic antiphospholipid syndrome: a review of pathogenesis, clinical features and treatment. PMID- 10804901 TI - The impact of new immigrants from the former Soviet Union on the severity of coronary angiographic findings in a public hospital in Israel. AB - BACKGROUND: The arrival of 610,000 new immigrants to Israel from the former Soviet republics accounted for 58% of the population growth in the early 1990's. OBJECTIVE: To compare the coronary angiographic findings and risk factors between the new immigrants and local Jewish and Arab patients in this era of cost containment. METHODS AND RESULTS: A total of 550 consecutive patients--314 Jews, 95 new immigrants and 141 Arabs--were catheterized and analyzed during a 5 month period in 1995. Of this group 403 were males (73%). The mean age was 63.6 +/- 10.2 years among new immigrants, 62.4 +/- 9.4 among Jews, and 55.1 +/- 10.9 among Arabs (P < 0.05). Immigrants, including those under age 60, had the highest prevalence of multivessel disease (88.7%). Arabs had a high prevalence of single vessel disease (34.6%) and a low prevalence of multivessel (65.4%) and left main coronary disease (5.6%). Age, gender, risk factors and ethnic origin in descending order were determinants of the extent of coronary angiographic disease as revealed by multiple regression analysis. CONCLUSION: New immigrants had the most extensive angiographic coronary involvement, while Arab patients were younger and had less severe coronary artery disease. More intensive risk factor modification may have a major impact on disease progression particularly in the new immigrant subgroup. PMID- 10804902 TI - Talismans and amulets in the Pediatric Intensive Care Unit: legendary powers in contemporary medicine. AB - BACKGROUND: For centuries talismans and amulets have been used in many cultures for their legendary healing powers. METHODS: We asked the parents of every child (Jews and Arabs) admitted to the Pediatric Intensive Care Unit over a 2 month period to complete a questionnaire, which included demographic data on the patient and the family, the use of talismans or other folk medicine practices, and the perception of the effects of these practices on the patient's well-being. A different questionnaire was completed by the ICU staff members on their attitude toward the use of amulets. RESULTS: Thirty percent of the families used amulets and talismans in the ICU, irrespective of the socioeconomic status of the family or the severity of the patient's illness. Amulets and talismans were used significantly more by religious Jews, by families with a higher parental educational level, and where the hospitalized child was very young. The estimated frequency of amulet use by the children's families, as perceived by the staff, was significantly higher than actual use reported by the parents. In Jewish families the actual use of amulets was found to be 30% compared to the 60% rate estimated by the medical staff; while in Moslem families the actual use was zero compared to the staff's estimation of about 36%. Of the 19 staff members, 14 reported that the use of amulets seemed to reduce the parents' anxiety, while 2 claimed that amulet use sometimes interfered with the staff's ability to carry out medical treatment. CONCLUSIONS: The use of talismans in a technologically advanced western society is more frequent than may have been thought. Medical and paramedical personnel dealing with very ill patients should be aware of the emotional and psychological implications of such beliefs and practices on patients and their families. PMID- 10804903 TI - Diminished chemokine and cytokine-induced adhesion of CD4+ T cells to extracellular matrix ligands in patients with end-stage renal failure. AB - BACKGROUND: Cell-mediated immunity is impaired in uremia. Cell-matrix interactions of immune cells such as CD4+ T lymphocytes with extracellular matrix are an important requirement for an intact immune response. The adherence of CD4+ T cells of healthy subjects (normal T cells) to ECM components is inhibited in the presence of uremic serum. Such decreased adhesive capacity is also found in T cells of dialysis patients. Various chemokines and cytokines affect the attachment of CD4+ T cells to ECM. OBJECTIVE: To evaluate chemokine (MIP-1 beta and RANTES) and tumor necrosis factor-alpha-induced adhesion of CD4+ T cells to ECM in a uremic milieu. METHODS: We examined adhesion of normal CD4+ T cells (resting and activated) to intact ECM in response to soluble or bound chemokines (MIP-1 beta and RANTES) and to TNF-alpha following incubation in uremic versus normal serum. Thereafter, we evaluated the adhesion of resting CD4+ T cells from dialysis patients in a similar fashion and compared it to that obtained from a healthy control group. RESULTS: Addition of uremic serum diminished soluble and anchored chemokine-induced attachment of normal resting and activated CD4+ T cells to ECM compared to a normal milieu (a peak response of 10-11% vs. 24-29% for soluble chemokines, P < 0.001; 12-13% vs. 37-39% for bound chemokines on resting cells, P < 0.01; and 18-20% vs. 45-47% for bound chemokines on activated cells, P < 0.02). The same pattern of response was noted following stimulation with immobilized TNF-alpha (7 vs. 12% for resting cells, P < 0.05; 17 vs. 51% for activated cells, P < 0.01). Adherence of dialysis patients' cells to ECM following stimulation with both bound chemokines was reduced compared to control T cells (15-17% vs. 25-32%, P < 0.0000). In contrast, adherence following stimulation by TNF-alpha was of equal magnitude. CONCLUSIONS: Abnormal adhesive capacity of T lymphocytes to ECM in uremia may, in part, be related to a diminished response to MIP-1 beta, RANTES and TNF-alpha. However, whereas reduced adhesion to chemokines was present in both normal CD4+ T cells in a uremic environment and in dialysis patients' T cells, TNF-alpha-induced adhesion was found to be inhibited only in normal cells in a uremic milieu. PMID- 10804904 TI - Prolactin serum level in patients with breast cancer. AB - BACKGROUND: Previous studies have suggested that prolactin may serve as an indicator of disease progression in breast cancer. OBJECTIVES: To evaluate the use of PRL as a serum tumor marker in patients with breast cancer. METHODS: PRL serum level was determined in 99 breast cancer patients and compared with CA 15-3 serum level. RESULTS: Elevated serum level of PRL (> 20 ng/ml) was found in 8 of 99 patients (8.1%). A stratified analysis of prolactin levels according to therapy revealed that PRL levels was increased in 8 of 55 untreated patients (14.5%), but not in patients who received hormonal or chemotherapy in the 3 months preceding the test (0/42 patients, P = 0.009). However, mean PRL level was similar in patients with no evidence of disease activity and in patients with active disease (10.2 vs. 8.2 ng/ml, NS). In comparison, CA 15-3 mean level was significantly lower in patients with no evidence of disease as compared to patients with active disease (18.2 vs. 144.7 units/ml, P < 0.001). PRL level was increased in 6 of 60 patients (10%) with no evidence of disease and in 2 of 39 (5.2%) with active disease (NS). In comparison, CA 15-3 level was increased in 3 of 60 patients (5%) with no evidence of disease and in 24 of 39 (61.5%) with active disease (P < 0.001). CONCLUSIONS: PRL levels are decreased following hormonal or chemotherapy in patients with breast cancer and there is no correlation between PRL serum level and the state of disease. Further studies are needed to clarify a possible clinical significance of hyperprolactinemia in a subset of patients with breast cancer. PMID- 10804905 TI - Autologous chondrocyte transplantation for reconstruction of isolated joint defects: the Assaf Harofeh experience. AB - BACKGROUND: Articular cartilage is incapable of undergoing self-repair since chondrocytes lose their mitotic ability as early as the first year of life. Defects in articular cartilage, especially in weight-bearing joints, will predictably deteriorate toward osteoarthritis. No method has been found to prevent this deterioration. Drilling of the subchondral bone can lead to fibrocartilage formation and temporary repair that slowly degrades. Animal experiments indicate that introducing proliferating chondrocytes such as cultured articular chondrocytes can reliably reconstruct joint defects. OBJECTIVES: To describe our clinical experience in culturing and transplanting autologous chondrocytes. METHODS: Biopsies were obtained from 10 patients, aged 18-45, undergoing a routine arthroscopy in which a cartilage defect was identified with indications for cartilage transplantation. The biopsies were further processed to establish chondrocyte cultures. ACT was performed in 8 of the 10 patients because of persistent symptoms for at least 2 months post-arthroscopy. All patients (6 men and 2 women) had a grade IV cartilage defect in the medial or lateral femoral condyle, and three had a defect in the trochlear region as well. Biopsies were removed from the lateral rim of the superior aspect of the femur, and cells were cultured in a clean room. Following a 2 order of magnitude expansion, cells were implanted under a periosteal flap. RESULTS: The eight patients implanted with autologous cells were followed for 6 months to 5 years (average 1 year). Complaints of giving-way, effusion and joint locking resolved in all patients, and pain as assessed by the visual analogue score was reduced by an average of 50%. Follow-up magnetic resonance imaging studies in all patients revealed that the defects were filled with tissue having similar signal characteristics to cartilage. CONCLUSIONS: Chondrocyte implantation is a procedure capable of restoring normal articular cartilage in cases with isolated joint defects. Pain can be predictably reduced, while joint locking and effusion are eliminated. The effect on osteoarthritis progression in humans has not yet been elucidated. PMID- 10804906 TI - Normalization of heart rate variability in post-traumatic stress disorder patients following fluoxetine treatment: preliminary results. AB - BACKGROUND: Spectral analysis of heart rate variability has been shown to be a reliable non-invasive test for quantitative assessment of cardiovascular autonomic regulatory responses, providing a window reflecting the interaction of sympathetic and parasympathetic tone. Alterations in autonomic function are associated with a variety of physiologic and pathophysiologic processes and may contribute substantially to morbidity and mortality. Our previous study shows that patients with post-traumatic stress disorder have significantly lower HRV compared to controls, reflecting a basal autonomic state characterized by increased sympathetic and decreased parasympathetic tone. OBJECTIVES: To apply this tool to PTSD patients treated with selective serotonin re-uptake inhibitors in order to assess the impact of such treatment on the autonomic dysregulation characterizing these patients. METHODS: Standardized heart rate analysis was carried out in nine PTSD patients treated with SSRI agents and compared to that in a matched control group of nine healthy volunteers and in nine untreated PTSD patients, based on a 15 minute resting electrocardiogram. RESULTS: Our preliminary results show that the HRV parameters indicating autonomic dysregulation, which characterize PTSD patients at rest, are normalized in responding patients by use of SSRIs. Neither the clinical implications of these findings nor their physiological mechanisms are clear at present, although we presume that they reflect a central effect, since the peripheral autonomic effects of SSRIs are relatively negligible. PMID- 10804907 TI - Extension of the organ pool in kidney transplantation: first year experience of the Israel Transplant Center. AB - BACKGROUND: Over a 12 month period, the Israel Transplant Center doubled the number of donors by assigning a nurse coordinator to each of 22 hospitals around the country and by using kidneys from elderly donors. OBJECTIVE: To evaluate the impact of our "marginal donors" policy on the results immediately following transplantation. METHODS: Between October 1997 and September 1998, 140 cadaveric kidney transplantations from 72 donors were performed in Israel. We defined two groups of recipients: patients with immediate graft function and patients with either delayed graft function requiring > 1 week of dialysis post-transplant or with primary graft non-function. We compared the following parameters between groups: donor and recipient age and gender, cause of donor's death, length of stay in the intensive care unit, vasopressor dosage and creatinine levels before harvesting, cold ischemic time, and the number of recipient grafts. RESULTS: There were 102 recipients (72.8%) with immediate graft function and 38 with either PNF (n = 13, 9.3%) or DGF (n = 25, 17.9%). On regression analysis, donor age > 50 year and retransplantation were significant risk factors for PNF or DGF (odds ratio 4.4 and 2.8, respectively). Of the 56 kidneys from donors > 50 years old, 21 (37.5%) developed either PNF (n = 9) or DGF (n = 12). CONCLUSIONS: We conclude that kidneys from donors over age 50 are at increased risk for graft non function or delayed function. Better assessment of functional capacity of kidneys from "aged" donors may help to choose appropriate donors from that pool. PMID- 10804908 TI - The unresolved problem of treating articular cartilage lesions. PMID- 10804909 TI - Rationing scarce cadaver organs: a medical responsibility? PMID- 10804910 TI - Flow cytometry application in organ transplantation. PMID- 10804911 TI - Constrictive pericarditis presented by generalized edema (anasarca). PMID- 10804912 TI - Scorpion envenomation and myocardial damage. PMID- 10804913 TI - The watermelon season. PMID- 10804914 TI - Uncommon sonographic appearance of a submental mass. PMID- 10804915 TI - Colorectal cancer is preventable. PMID- 10804916 TI - "Buttoned up". PMID- 10804918 TI - 39th Interscience Conference on Antimicrobial Agents and Chemotherapy, 26-29 September 1999, San Francisco. PMID- 10804917 TI - Bedside humanities. A vision from the renaissance and two case reports from the present. PMID- 10804919 TI - A 54-year-old woman was admitted to the emergency unit because of pains in the right lower abdomen. PMID- 10804920 TI - Lucid and comprehensive review of the approach to acute bronchiolitis in Israel. PMID- 10804921 TI - Children with type I Gaucher disease: growing into adulthood with and without enzyme therapy. PMID- 10804922 TI - The race for the diagnostic autoantibody in celiac disease. And the winner is... PMID- 10804923 TI - The diagnostic and prognostic importance of neonatal length measurements. PMID- 10804924 TI - Induction of mRNA for tumor necrosis factor-alpha and interleukin-1 beta in mice brain, spleen and liver in an animal model of Shigella-related seizures. AB - BACKGROUND: The pathogenesis of neurological symptoms, the most common extraintestinal complication of childhood shigellosis, is unclear. To elucidate the mechanisms involved, we developed an animal model and demonstrated that TNF alpha and IL-1 beta play a role. OBJECTIVES: To determine whether TNF alpha and IL-1 beta genes are expressed in the brain following peripheral administration of Shigella dysenteriae 60R. METHODS: Expression of mRNA for TNF alpha and IL-1 beta was examined in the brain structures (hypothalamus and hippocampus) and peripheral organs by reverse transcriptase polymerase chain reaction, at different time points after intraperitoneal injection of S. dysenteriae sonicate. RESULTS: In our animal model of Shigella-related seizures, TNF alpha and IL-1 beta mRNA were induced in the brain, spleen and liver already 1 hour after injection of S. dysenteriae sonicate. The expression of TNF alpha and IL-1 beta mRNA in spleen, hippocampus and hypothalamus decreased after 6 h and increased again at 18 h post-injection. CONCLUSIONS: Local production of TNF alpha and IL-1 beta in the brain may be involved in the enhanced seizure response of mice after administration of S. dysenteriae. It is possible that intracerebral production of TNF alpha and IL-1 beta plays a role in neurological disturbances of human shigellosis. PMID- 10804925 TI - The effect of cisapride on total parenteral nutrition-associated cholestasis in rats. AB - BACKGROUND: Cholestasis is a frequent problem in patients on total parenteral nutrition. Cisapride has a prokinetic effect on the biliary system, but its effect on hepatic excretory function is unknown. OBJECTIVES: To study the effect of cisapride on TPN-induced cholestasis in a rat model. METHODS: Bile flow and bile salt secretion rate were measured in rats given TPN. There were four groups of 8 to 13 animals each. After a one hour baseline period during which all four groups received i.v. saline infusion, two groups received a TPN solution for another 2 hours, while saline was infused in the two control groups. At the beginning of the second hour, 2 mg/kg cisapride was injected i.v. as a bolus into one experimental and one control group. Bile was collected from the common bile duct. RESULTS: At the end of the third hour, TPN caused a significant reduction in bile flow (P < 0.02) and bile salt secretion rate (P < 0.001) (61.24 vs. 50.74 microliters/min/kg, and 1.173 vs. 0.799 mumol/min/kg, respectively). Addition of cisapride abolished the cholestatic effect of TPN. CONCLUSIONS: Cisapride has a protective effect against TPN-associated cholestasis. This may have clinical significance, and further studies are warranted. PMID- 10804926 TI - Reaching the diagnosis of cystic fibrosis--the limits of the spectrum. AB - BACKGROUND: Cystic fibrosis is the most common life-limiting autosomal recessive genetic disorder in Caucasians. Typically it is a multisystem disease diagnosed by increased chloride levels on sweat testing, with mortality due mainly to progressive respiratory disease. The clinical spectrum of CF has recently been much expanded. Genetic testing for mutant CF transmembrane regulator has revealed atypical cases where sweat test results are borderline or normal. In other patients, genetic mutations cannot be identified but abnormal CFTR function is shown using nasal potential difference measurement. OBJECTIVES: To highlight the diagnostic and therapeutic dilemmas in cases of atypical cystic fibrosis. METHODS: We reviewed patients with atypical CF and widely varying phenotype who are managed at Schneider Children's Medical Center of Israel. RESULTS: Two patients had severe lung disease but little expression in other organs. Accurate diagnosis was essential to enable aggressive therapy in a specialized center. Four other patients are in excellent general health but have symptoms limited to male infertility, heat exhaustion, pancreatitis or transient liver dysfunction, while lung disease is minimal. For these patients, careful counseling is needed to avoid unnecessary upheaval, inappropriately aggressive management, and the psychosocial implications of a CF diagnosis. These dilemmas have increased considerably in our center, as in others worldwide. CONCLUSION: It is our obligation as clinicians--at the level of both primary physician and referral center--to maintain an ever higher index of suspicion for CF, tempered by a rational program of counseling and management appropriate to the individual. PMID- 10804927 TI - Exercise in childhood asthma: provoking agent, diagnostic tool and therapeutic measure. PMID- 10804928 TI - Soluble CD14 in children with status asthmaticus. AB - BACKGROUND: Inflammation is a major component in the pathogenesis of asthma. CD14 is an endotoxin (lipopolysaccharide) receptor, and is expressed mainly on monocytes and macrophages. Binding of LPS to CD14 activates the monocyte or macrophage and causes the release of different cytokines. The soluble form of CD14 is present in serum, and its concentration increases in several clinical conditions, including infections, autoimmune disorders, allergic disorders, and lung diseases. The possible role of CD14/sCD14 in asthma has been investigated in a few adult patients only. OBJECTIVES: To measure serum concentrations of sCD14 in children with status asthmaticus. METHODS: We compared serum concentration of sCD14 in 10 children with status asthmaticus measured within 24 hours of admission and after recovery from the acute episode. RESULTS: Levels of sCD14 were significantly higher during acute asthma attacks than at recovery. CONCLUSIONS: The elevated serum levels of sCD14 during status asthmaticus may be the result of the activation of monocytes, macrophages or other cells. The influence of medications on serum sCD14 cannot be ruled out. The possible use of sCD14 as a marker of lung inflammation in asthma warrants further investigation. PMID- 10804929 TI - The diagnostic and therapeutic approach to acute bronchiolitis in hospitalized children in Israel: a nationwide survey. AB - BACKGROUND: Bronchiolitis caused by respiratory syncytial virus is one of the major causes of hospitalization in young children, especially during the winter. Recent evidence has shown that pharmacological treatment, especially nebulized epinephrine, in addition to the traditional supportive treatment, can alleviate symptoms and shorten hospitalization, but this approach is not yet widespread. OBJECTIVES: To determine whether the management of bronchiolitis in Israel is moving toward a stronger emphasis on pharmacological care. METHODS: A questionnaire on the diagnosis and management of bronchiolitis was completed by 27 heads of pediatric departments throughout Israel. The questionnaire dealt with the frequency of usage of diagnostic and selected therapeutic procedures. RESULTS: Chest X-ray and arterial blood gases are commonly used as a diagnostic aid in more than 75% of the departments, and antibiotics are prescribed routinely in 24%. Corticosteroids are still in use: 48% use systemic steroids, and 19% nebulized steroids. Nebulized epinephrine is used in 22% of the departments, while nebulized beta-agonists are used frequently in two-thirds of the departments. CONCLUSIONS: Despite convincing data that beta-agonists and steroids have no positive effect on the outcome of bronchiolitis on the one hand, and that nebulized epinephrine has advantages in children on the other, we found significant use of the former two agents and sparse use of the latter. Greater awareness is needed among pediatricians, and measures should be introduced to incorporate the new recommendations, with further study of the effect of the old and new drugs on bronchiolitis. PMID- 10804930 TI - Anomalous origin of the left coronary artery from the pulmonary artery: diagnosis and postoperative follow up. AB - BACKGROUND: Anomalous origin of the left coronary artery from the pulmonary artery is a rare congenital malformation that presents a diagnostic challenge to the pediatrician and pediatric cardiologist. Although surgical repair is always indicated, the optimal technique has yet to be determined. OBJECTIVES: To review our experience with the diagnosis of children with ALCAPA and to assess short to midterm surgical results. METHODS: Between 1992 and 1998, 13 infants and children (2 months to 15 years) were treated for ALCAPA at our medical center. Eight were diagnosed during the first year of life; all were symptomatic and had severe dysfunction of the left ventricle. The five patients diagnosed at an older age had normal myocardial function. Diagnosis was established by echocardiography alone in seven patients; six required catheterization (one infant and all older patients). Surgery was performed in 12 patients to establish dual coronary artery system: 7 underwent the Takeuchi procedure and 5 had re-implantation of the anomalous left coronary artery. RESULTS: One infant died shortly after diagnosis before surgical repair was attempted, and one died postoperatively. Four patients required additional surgery: three for late complications of the Takeuchi procedure and one valve replacement for mitral insufficiency. Recent evaluation revealed good global left ventricle function in all patients except for one, who is still within the recovery phase and shows gradual improvement. However, most patients who presented with severe myocardial dysfunction upon diagnosis still display abnormal features such as echo-dense papillary muscles or evidence of small akinetic segments. In this group, early repair was associated with faster myocardial recovery. CONCLUSIONS: The diagnosis of ALCAPA remains a clinical challenge to the pediatrician and cardiologist. Diagnosis can be established echocardiographically, and early diagnosis and treatment may lead to faster myocardial recovery. The preferred surgical method appears to be re-implantation of the ALCA. The chance for good recovery of global ventricular function is high even in the sickest patients, nonetheless abnormal myocardial features can be identified even years after surgery. PMID- 10804931 TI - The Ross operation: initial Israeli experience. AB - BACKGROUND: The need for aortic valve replacement in children and young adults poses a special problem to cardiologists and surgeons. Replacing the sick aortic valve with the patient's pulmonary valve as described by Ross has proven to be a good option in this special age group. OBJECTIVE: To review our initial experience in order to assess the short-term results. METHODS: From January 1996 to June 1999, 40 patients (age 8 months to 41 years) underwent aortic valve replacement with pulmonary autograft. Indications for surgery were congenital aortic valve disease in 30 patients, bacterial endocarditis in 5, rheumatic fever in 3, and complex left ventricular outflow tract obstruction in 3. Trans esophageal echocardiography was performed preoperatively and post-bypass in all patients, and transthoracic echocardiography was done prior to discharge and on follow-up. RESULTS: There was no preoperative or late mortality. All patients remain in functional class I (New York Heart Association) and are free of complications and medication. None showed progression of autograft insufficiency or LVOT obstruction. Homograft insufficiency in the pulmonary position has progressed from mild to moderate in one patient, and three developed mild homograft stenosis. CONCLUSIONS: The Ross procedure can be performed with good results in the young population and is considered an elegant surgical alternative to prosthetic values and homografts. PMID- 10804932 TI - Blood pressure measurements in Israeli Arab children and adolescents. AB - BACKGROUND: Hypertension is one of the most prevalent vascular diseases in the adult population. It is an important determinant of atherosclerosis in adolescents and young adults. There is to date no information on blood pressure in children of the Israeli Arab population. OBJECTIVES: To study blood pressure in Israeli Arab children and adolescents. METHODS: Blood pressure measurements were taken in the supine position in 4,488 Israeli Arab children and adolescents of both sexes aged 6-17 years. Height and weight were also determined. Correlation was investigated between systolic and diastolic blood pressure, body mass index, gender, and age. RESULTS: The systolic and diastolic blood pressures were similar in both sexes for all age groups up to 14 years of age. Systolic blood pressure was significantly (P < 0.001) higher in males aged 15-17 years (120 +/- 13 vs. 111 +/- 12 mmHg, 123 +/- 13 vs. 113 +/- 14.0 mmHg, and 123 +/- 18 vs. 111 +/- 14 mmHg, respectively). Diastolic blood pressure was higher in males aged 15-17, with a statistically significant difference for age 17 only (75 +/- 12 vs. 69 +/- 13 mmHg). Blood pressure was elevated in 322 students in the initial screening (7.17%), with a decrease to 2.18% when this group was rescreened 2 weeks later. The systolic blood pressure in our group is higher than that in Jewish Israeli children of Asian and North African origin, and in American children. It is similar to the systolic blood pressure of European children and Jewish children born in Israel. The diastolic blood pressure in our group is higher than that in all groups of Israeli Jewish children and American children of different ethnic groups. CONCLUSIONS: Israeli Arab children and adolescents have higher blood pressure levels than their Israeli Jewish counterparts. Further studies are required to confirm this observation. PMID- 10804933 TI - Comparison of assays for anti-endomysial and anti-transglutaminase antibodies for diagnosis of pediatric celiac disease. AB - BACKGROUND: Anti-endomysial antibodies are sensitive and specific markers for celiac disease. This antibody has recently been identified as an antibody to tissue transglutaminase, an enzyme that cross-links and stabilizes extracellular matrix proteins. OBJECTIVES: To evaluate the clinical usefulness of an enzyme linked immunoassay for anti-transglutaminase antibodies, and to compare the results with those of AEA, the current gold standard serological test for celiac disease. METHODS: Serum samples were collected from 33 patients with biopsy proven celiac disease and AEA tests were performed. Control samples for anti transglutaminase were obtained from 155 patients. An ELISA test for immunoglobulin A anti-transglutaminase utilizing guinea pig liver transglutaminase was developed and performed on all sera. Cutoff values for the test were performed using logistic regression and receiver operating curves analysis. RESULTS: An optical density cutoff value of 0.34 was established for the assay. The mean value was 0.18 +/- 0.19 optical density for controls, and 1.65 +/- 1.14 for patients with celiac disease (P < 0.001). Sensitivity and specificity of the assay were both 90%, while AEA had a sensitivity and specificity of 100% and 94%, respectively. CONCLUSIONS: A tissue transglutaminase based ELISA test is both sensitive and specific for detection of celiac disease. PMID- 10804934 TI - The role of Helicobacter pylori and gastritis in children with recurrent abdominal pain. AB - BACKGROUND: Recurrent abdominal pain is a common pediatric diagnostic problem. Endoscopy is sometimes performed as part of the evaluation. Although gastritis and/or Helicobacter pylori infection is often present, it is not known if they contribute to the symptomatology. OBJECTIVES: To evaluate the role of either gastritis or H. pylori infection in the symptomatology of children with RAP. PATIENTS AND METHODS: We retrospectively studied two groups of patients, 70 children in each, who had undergone endoscopy. One group was evaluated endoscopically for RAP and the other was a heterogeneous group that underwent endoscopy for indications other than RAP. Biopsies were taken during endoscopy and Giemsa staining was performed for the presence of H. pylori. Triple therapy was given as indicated, and the children were followed for an average of 6 months. RESULTS: Microscopic gastritis was diagnosed in 39 patients (55.7%) of the RAP group and in 31 of the heterogeneous group (44.2%) (NS), and H. pylori was found in 32 patients of the RAP group and in 16 of the heterogeneous group (45.7% vs. 22.8%, P < 0.01). All children with H. pylori, except one in the heterogeneous group, had accompanying gastritis. On the other hand, gastritis without H. pylori infection was seen in 7 children in the RAP group and in 15 of the other. Endoscopy revealed macroscopic abnormalities in 52 of the 70 children with microscopic gastritis. There was a clinical improvement after triple therapy in 28 of 33 children with H. pylori-associated gastritis (84.85%), in 4 of 8 children with gastritis unassociated with H. pylori (50%), and in 8 of 15 without gastritis or H. pylori (53.3%) (P < 0.01 between the H. pylori-associated gastritis and each of the other groups). CONCLUSIONS: H. pylori infection and gastritis may be associated with RAP in a selected subgroup of children. We recommend a complete work-up, including endoscopy and invasive or non-invasive diagnostic modalities for H. pylori, and treatment of the infection. PMID- 10804935 TI - Small bowel obstruction and covered perforation in childhood caused by bizarre bezoars and foreign bodies. AB - BACKGROUND: Small bowel obstruction with perforation is an unusual and rare complication of bezoars. OBJECTIVE: To describe our use of emergency laparotomy to treat intestinal obstruction caused by bizarre bezoars. CONCLUSIONS: An aggressive surgical approach to intestinal obstruction in the pediatric disabled or mentally retarded population is recommended. PMID- 10804937 TI - Secondary enuresis: post-traumatic stress disorder in children after car accidents. AB - BACKGROUND: In our experience, secondary enuresis nocturna is a common complaint among children after a motor vehicle accident. However, as these children are often brought for examination as part of an insurance compensation claim, this complaint is not always reliable. OBJECTIVE: To describe a series of children in whom secondary enuresis occurred after a motor vehicle accident. METHODS AND RESULTS: Five children were brought to our clinic for evaluation of secondary nocturnal enuresis. Review of past history revealed a car accident preceding the onset of the enuresis. All but one had additional behavioral symptoms typical of post-traumatic stress disorder. Four children had evidence of head trauma, and one had psychological but no physical trauma. CONCLUSIONS: Nocturnal enuresis can occur after a motor vehicle accident due either to purely psychological trauma or organic head trauma. While nocturnal enuresis is generally attributed to organic causes, psychological mechanisms also play a significant role. PMID- 10804936 TI - Traumatic adrenal injury in children. AB - BACKGROUND: Multiple organ injury in children is an increasingly frequent phenomenon in the modern emergency room. Adrenal hemorrhage associated with this type of trauma has received little attention in the past. OBJECTIVES: Using computed tomography, we sought to determine the rate and nature of adrenal gland injury in children following blunt abdominal trauma due to motor vehicular accident. METHODS: A total of 121 children with blunt abdominal trauma were examined and total body CT was performed in cases of multi-organ trauma or severe neurological injury. RESULTS: Of all the children who presented with blunt abdominal trauma over a 51 month period, 6 (4.95%) had adrenal hemorrhage. In all cases only the right adrenal gland was affected. Coincidental injury to the chest and other abdominal organs was noted in 66.7% and 50% of patients, respectively. CONCLUSIONS: Traumatic adrenal injury in the pediatric population may be more common than previously suspected. Widespread application of the more sophisticated imaging modalities available today will improve the detection of damage to the smaller organs in major collision injuries and will help in directing attention to the mechanism of trauma. PMID- 10804938 TI - Profile of the pediatric burn patient at the Schneider Children's Medical Center of Israel. AB - BACKGROUND: Burn trauma occurs mostly in young children. Burn injury in the pediatric age group has multiple-aspect sequelae. OBJECTIVES: To characterize the profile of the injured pediatric burn patient, thus targeting the most vulnerable pediatric group. METHODS: Between 1 January and 31 December 1996, a total of 9,235 pediatric patients were admitted for various traumatic injuries (burns, lacerations, fractures, etc.) to the Emergency Medicine Department of Schneider Children's Medical Center. We conducted a retrospective study of the patients' charts, including demographic data, which were stored in a computerized database, for statistical evaluation. The characteristics of pediatric burn patients were examined and compared with other pediatric trauma patients. RESULTS: Of the total patient population, 282 (3.1%) suffered from burns (37% females, 63% males). The most frequent burn injury was scald burn (58%). The pediatric group that was most exposed to burns was 13-18 month old males. CONCLUSIONS: Having identified the high risk group among the pediatric burn patients, we suggest that prevention programs be directed towards this group in order to reduce further risk of burn injury. PMID- 10804940 TI - Erythema nodosum in Israeli children. AB - BACKGROUND: Erythema nodosum, although uncommon in children, is the most frequent form of panniculitis in pediatrics. EN has been associated with various infections and chronic inflammations, and its course varies with age, gender, and racial and geographic factors. There is no information on EN in Israeli children. OBJECTIVES: To examine the clinical course of EN and the conditions with which it is associated in Israeli children. METHODS: We conducted a retrospective study of 24 children with a diagnosis of EN who presented at our Center over a 10 year period (1989-98). RESULTS: EN was more frequent in females than males (ratio 2:1) due to a cluster of adolescent girls. The mean duration of the skin manifestation was 18 days. The course was benign in all patients. Streptococcal infection was the most common cause (25%), followed by Epstein-Barr virus infection (18%) and inflammatory bowel disease (13%). In one-third of cases, no specific cause could be identified. Tuberculosis, an important cause of EN in the past, was not found in our patients. CONCLUSIONS: Most cases of EN in Israeli children are related to streptococcal and EBV infections or to chronic inflammatory conditions. Despite the increase in tuberculosis morbidity in Israel during recent years, we found no association of EN and tuberculosis in our study. PMID- 10804939 TI - Poison exposure in children before Passover. AB - BACKGROUND: Extensive cleaning of homes in Israel before Passover may result in increased exposure of children to cleaning substances. OBJECTIVES: To evaluate the potential danger of Passover cleaning to children, and to study the risk factors in order to identify areas for prevention. METHODS: All cases of poison exposure in Jewish and Arab children under the age of 15 years reported to the Israel Poison Information Center during 1990-95 (n = 5,583) were analyzed for the 6 weeks before and 6 weeks after Passover. Poison exposures in Jewish children < 15 years old were studied in seven pediatric emergency rooms for the 2 weeks before and 6 weeks after Passover (n = 123). RESULTS: The IPIC data showed a highly significant 38% increase in the average weekly poison exposure rate for the 2 weeks before Passover compared with the remaining 10 weeks. Data recorded by the pediatric emergency rooms showed a twofold increase in cleaning substance poisoning during the 2 weeks before Passover compared with the following 6 weeks. The rise in exposures to cleaning substances was observed among children from secular, religious and ultra-orthodox families. In these exposures, the substance was found in open containers in 70% of cases. CONCLUSIONS: The extensive cleaning of homes among Jewish families in preparation for Passover poses the danger to young children of cleaning substance poisoning. Increasing public awareness, closer observation of children, and keeping these substances in closed containers should increase children's safety during this annual cleaning. PMID- 10804941 TI - Procalcitonin correlates with C-reactive protein as an acute-phase reactant in pediatric patients. AB - BACKGROUND: Previous reports on the behavior of procalcitonin blood levels in diverse clinical conditions suggest that it is part of the activation of cellular immunity and is another acute-phase reactant. OBJECTIVE: To compare procalcitonin with C-reactive protein, a well-known acute-phase reactant, in a series of acutely febrile pediatric patients and to review recent literature on procalcitonin. METHODS: Procalcitonin and CRP levels were evaluated in 38 blood samples of pediatric patients who were admitted to the Dana Children's Hospital for evaluation of unexplained fever or for sepsis work-up. RESULTS: The parallelism between procalcitonin and CRP was found to be highly significant (P < 0.01). CONCLUSION: The rise of procalcitonin blood levels in febrile pediatric patients suggests that it is part of the acute-phase reaction, parallel with the CRP reaction. PMID- 10804942 TI - Osteoid osteoma: resection with CT guidance. AB - BACKGROUND: Osteoid osteoma is a benign bone lesion characterized by nocturnal pain mostly, which may be relieved by non-steroidal prostaglandin inhibitors. Treatment by complete resection of the nidus immediately relieves the pain. Intraoperative location of the nidus may be difficult, and extensive bone resection may be necessary to ensure complete excision. Few studies have described resection of osteoid osteoma under CT guidance, and little attention has been given to lesions near the neurovascular bundle. OBJECTIVE: To report our results of osteoid osteoma resection under CT guidance, with specific attention to lesions lying near the neural structure. METHODS: Nine patients with suspected osteoid osteoma underwent resection with a 6.8 mm core drill under CT guidance. RESULTS: Histologic confirmation was obtained in seven patients, while in two there was no evidence of the nidus in the excised bone material. All nine reported complete pain relief immediately after the surgery. Postoperative CT scan showed complete removal of the osteoid osteoma. CONCLUSIONS: Removal of osteoid osteoma under CT guidance is simple, safe and allows complete removal of the nidus with low morbidity. PMID- 10804943 TI - The prevalence of specific IgE antibodies to natural rubber latex in healthcare workers in Israel. AB - BACKGROUND: According to studies from different countries, the prevalence of natural rubber latex sensitization in healthcare workers ranges from 2.9 to 17%. OBJECTIVE: To estimate the prevalence of NRL-specific IgE antibodies in healthcare workers in Israel. METHODS: Three hundred healthcare workers, mostly from a major pediatric tertiary care facility, and 15 non-healthcare workers completed a questionnaire on signs and symptoms of NRL allergy and other respiratory and food allergies. NRL-specific IgE antibodies were assayed with the DPC AlaSTAT-ELISA method. RESULTS: Seventy of the 300 workers (23.3%) reported symptoms of NRL allergy: hand eczema and pruritus in 63, upper respiratory tract and ocular symptoms in 10, shortness of breath in 2, and generalized rash in 6. None had anaphylaxis due to latex exposure. There was a significant correlation of symptoms of NRL allergy with atopy and job category (nurses, laboratory technicians, nurse assistants and dental medicine workers), but not with gender, age, or years of employment. The in vitro tests for specific IgE antibodies against NRL were positive (> or = 0.70 IU/ml) in five workers (1.66%). CONCLUSIONS: This is the first study of the prevalence of NRL-specific IgE antibodies in healthcare workers in Israel. Our 1.66% sensitization rate is much lower than that reported for healthcare workers in other countries. This difference may be due to our inclusion of a study population with a relatively low exposure to latex gloves (pediatricians compared to surgeons). Further studies are needed in this and other high risk populations in Israel. PMID- 10804944 TI - Delayed growth and puberty in patients with Gaucher disease type 1: natural history and effect of splenectomy and/or enzyme replacement therapy. AB - BACKGROUND: Growth retardation in childhood was only recently recognized as a prominent feature of Gaucher disease type 1, but there are few data on both the pubertal development and the final outcome of growth and sexual maturation. OBJECTIVE: To investigate the natural pattern of growth and puberty in patients with Gaucher disease type 1 and the effect of splenectomy and enzyme replacement therapy. METHODS: We retrospectively analyzed growth and puberty in 57 patients with Gaucher disease type 1; 52 were followed since childhood and/or prepuberty and 42 have reached sexual maturity and final height. In the analysis we considered severity of disease, time of splenectomy, and start of enzyme replacement therapy. RESULTS: Deceleration of growth at age 3-5 years was observed in 30 of 57 patients followed since early childhood while untreated: height-SDS decreased from -0.34 +/- 0.42 at age 0-3 years to -1.93 +/- 0.95 (P < 0.01) at age 7-10 years and was more pronounced with severe disease. A high prevalence (59.6%) of delayed puberty, which was more frequent with severe disease, was observed in 47 patients followed before and throughout puberty. No primary endocrine pathology was found. All patients, untreated as well as treated, with growth and pubertal delay had a spontaneous catch-up, achieved full sexual maturation, and most (83.3%) reached a final height within the range of parental height-standard deviation score. Splenectomy (partial and/or total) performed in 20 patients while still growing had a beneficial effect on growth, which was temporary in some and did not affect puberty. ERT improved growth in 11 patients who started therapy before puberty, as evidenced by a progressive increase in the height-SDS, and seemed to normalize the onset of puberty. CONCLUSIONS: Growth retardation in childhood and delay of puberty are characteristic of Gaucher disease type 1 and are more frequent with severe disease. There is a spontaneous catch-up later in life and most patients reach a final height within their genetic growth potential. Enzyme replacement therapy apparently normalizes growth and possibly also the onset of puberty. PMID- 10804945 TI - Chronic hepatitis B virus in children in Israel: clinical and epidemiological characteristics and response to interferon therapy. AB - OBJECTIVE: To describe the clinical and epidemiological features of hepatitis B virus infection in Israeli children, and to evaluate their response and compliance to therapy. METHODS: We retrospectively studied 51 patients (34 males, 17 females), aged 2-18 years, from several medical centers in Israel. RESULTS: Of the 51 patients, 38 with elevated transaminase, positive hepatitis B e antigen and/or HBV DNA, and histologic evidence of liver inflammation were treated. Interferon was administered by subcutaneous injections three times a week for 3 12 months (dosage range 3-6 MU/m2). Only 16% were native Israelis, while 78% of the children were of USSR origin. A family history of HBV infection was recorded in 25 of the 51 patients (9 mothers, 16 fathers or siblings). Five children had a history of blood transfusion. The histological findings were normal in 3 patients, 24 had chronic persistent hepatitis, 14 had chronic active hepatitis and 2 had chronic lobular hepatitis. Five children also had anti-hepatitis D virus antibodies. Twelve of the 38 treated patients (31.5%) responded to IFN completely, with normalization of the transaminase levels and disappearance of HBeAg and HBV DNA. In no patient was there a loss of hepatitis B surface antigen. The main side effects of IFN were fever in 20 children, weakness in 10, headaches in 9, and anorexia in 6; nausea, abdominal pain, and leukopenia were present in 3 cases each. The response rate was not affected by age, country of origin, alanine/aspartate aminotransferase levels, or histological findings. However, a history of blood transfusion was a predictor of good response, 60% vs 27% (P < 0.05). CONCLUSIONS: We found IFN to be a safe and adequate mode of treatment in children with chronic HBV infection, regardless of their liver histology and transaminase levels. Therefore, in view of the transient side effects associated with this drug, we recommend considering its use in all children with chronic hepatitis B. PMID- 10804946 TI - Evaluation of the approach of primary care physicians to the management of streptococcal pharyngotonsillitis. IPROS Network. AB - BACKGROUND: Streptococcal pharyngotonsillitis remains a common illness in children and can lead to serious complications if left untreated. OBJECTIVE: To evaluate the diagnostic and management approach of a sample of primary care physicians in the largest sick fund in Israel to streptococcal pharyngotonsillitis in children. METHODS: A questionnaire was mailed to all physicians who treat children and are employed by the General Health Services (Kupat Holim Klalit) in the Jerusalem District. The questionnaire included data on demographics, practice type and size, and availability of throat culture and rapid strep test; as well as a description of three hypothetical cases followed by questions relating to their diagnosis and treatment. RESULTS: Of the 188 eligible physicians, 118 (62.5%) responded, including 65 of 89 pediatricians (73%) and 53 of 99 family and general practitioners (53.5%). Fifty-six physicians (47.4%) had more than 18 years experience, and 82 (70%) completed specialization in Israel. Mean practice size was 950 patients. Fifty-three physicians (43%) worked in Kupat Holim community clinics, 25 (21%) worked independently in private clinics, and 40 (34%) did both. A total of 91 (77%) had access to laboratory facilities for daily throat culture. The time it took for the results to arrive was 48 to 72 hours. For the three clinical scenarios, 90% of the physicians accurately evaluated case A, a 1-year-old with viral pharyngotonsillitis, and 100 (85%) correctly diagnosed case C, a 7-year-old with streptococcal infection. As expected, opinions were divided on case B, a 3-year-old child with uncertain diagnosis. Accordingly, 75 (65.3%) physicians did not recommend treatment for case A, compared to 109 (92.5%) for case C. For case B, 22 (19%) said they would always treat, 43 (36%) would sometimes treat, and 35 (30%) would await the result of the throat culture. For 104 (88%) physicians the antibiotic of choice for case C was penicillin, while only 9 (7.5%) chose amoxicillin. However, the recommended dosage regimens varied from 250 to 500 mg per dose, and from two to four doses daily. For case C, 110 physicians (93%) chose a 10 day duration of treatment. CONCLUSIONS: The primary care physicians in the sample (pediatricians, general practitioners and family physicians) accurately diagnosed viral and streptococcal pharyngotonsillitis. However, there was a lack of uniformity regarding its management in general, and the dosage regimen for penicillin in particular. PMID- 10804947 TI - Seroprevalence of pertussis antibodies among adolescents in Israel. AB - BACKGROUND: There is an increasing number of reports of pertussis among older children and adults. The development and licensure of an acellular pertussis vaccine offer the possibility of adult vaccination against the disease. Information on immunity to pertussis in this age group is needed before any vaccination policy can be considered. OBJECTIVES: To study the seroepidemiology of pertussis antibodies in a random sample of adolescents. METHODS: Serum IgG antibodies to whole-cell lysate of Bordetella pertussis were measured by enzyme linked immunosorbent assay in sera of 533 Israeli military recruits aged 17-18 years. Epidemiologic variables were collected by a questionnaire and analyzed for correlation with pertussis antibodies. RESULTS: Of the sera tested 58.6% were positive for pertussis IgG antibodies, while 35.4% were negative and 6% were borderline. The seropositivity rate was significantly higher among females and non-smokers than among males and smokers. Serum samples of subjects found negative to Bordetella pertussis on recruitment were tested again, using the same ELISA assay, 2-3 years later. Seroconversion during the 3 year military service was detected in 12.5% of 40 subjects. Using the pertussis toxin as the antigen in a subsample of 160 sera, the seroprevalence was lower than that detected by the whole-cell lysate on the same sera (45% vs. 58%). CONCLUSIONS: A significant part of the adolescent population in Israel has low titer of serum IgG antibodies to the multiple antigens of B. pertussis. The relatively low concentration of anti pertussis antibodies, together with the serological evidence of exposure to the disease indicates that booster immunization with the acellular pertussis vaccine of military recruits should be considered after more information on the incidence of clinical cases of pertussis will be available. PMID- 10804948 TI - Pregnancy outcome in women with primary renal disease. AB - BACKGROUND: The preconception and intraconception parameters that are relevant to outcome in women with underlying renal disease remain controversial. OBJECTIVES: To analyze the types and frequencies of short- and long-term (2 years after delivery) maternal and neonatal complications in 38 patients with primary renal disease (46 pregnancies), most of them with mild renal insufficiency. METHODS: Logistic regression models were formulated to predict successful outcome. RESULTS: Successful pregnancy outcome (live, healthy infant without severe handicap 2 years after delivery) was observed in 98% of the patients with primary renal disease. Factors found to be significantly predictive of successful outcome were absence of pre-existing hypertension, in addition to low preconception serum uric acid level. CONCLUSIONS: Most women with primary renal disease who receive proper prenatal care have a successful pregnancy outcome. Worse pregnancy outcome was observed in women with moderate or severe renal failure. Fitted logistic models may provide useful guidelines for counseling women with preexisting renal disease about their prospects for a successful pregnancy in terms of immediate and long-term maternal and neonatal outcome. PMID- 10804949 TI - Type IIIc Gaucher disease and acrodysostosis. PMID- 10804950 TI - Allyl isothiocyanate. PMID- 10804951 TI - ortho-anisidine. PMID- 10804952 TI - Atrazine. PMID- 10804953 TI - Butyl benzyl phthalate. PMID- 10804954 TI - Chloroform. PMID- 10804955 TI - Chlorothalonil. PMID- 10804956 TI - Cyclamates. PMID- 10804957 TI - Dichlorobenzenes. PMID- 10804958 TI - Hexachlorobutadiene. PMID- 10804959 TI - Hexachloroethane. PMID- 10804960 TI - d-limonene. PMID- 10804961 TI - Melamine. PMID- 10804962 TI - Methyl tert-butyl ether. PMID- 10804963 TI - Nitrilotriacetic acid and its salts. PMID- 10804964 TI - Paracetamol. PMID- 10804965 TI - ortho-phenylphenol and its sodium salt. PMID- 10804966 TI - Potassium bromate. PMID- 10804967 TI - Quercetin. PMID- 10804968 TI - Saccharin and its salts. PMID- 10804969 TI - Simazine. PMID- 10804970 TI - Surgical implants and other foreign bodies. PMID- 10804971 TI - Techniques for covering soft tissue defects resulting from plantar ulcers in leprosy: Part III. Use of plantar skin or musculocutaneous flaps and anterior leg flap. AB - Anatomical studies suggest that five types of plantar flaps namely, the lateral and medial plantar flaps, the Abductor hallucis-, the Flexor digitorum brevis-, and the Abductor digiti minimi-myocutaneous flaps, can be incised from the central section of the sole. The advantages of a plantar flap are recognizable neurovascular bundles of the sole, wide calibre of constantly located blood vessels, identical histological structure of the donor and the recipient sites, hidden donor site and absence of functional deficit. We have used the plantar flaps in seven cases. There has been no recurrence of ulceration in any of them during the follow-up period of 12 to 108 months. An anterior leg flap based on the cutaneous branches of the anterior tibial artery, with firmly anchored vessels, a long pedicle with wide vessels may be used not only as a free flap graft for reconstruction of moderate degree distant defects but also as a retrograde island flap graft for the reconstruction of adjacent tissue defect. We have used the retrograde island flap graft based on the anterior tibial artery in five cases of plantar ulceration with satisfactory results. There was no recurrence of ulceration during the follow-up period of 48 to 72 months. PMID- 10804972 TI - Techniques for covering soft tissue defects resulting from plantar ulcers in leprosy: Part IV. Use of medial leg flap and medial knee flap. AB - The medial leg flap, based on the cutaneous branches of the posterior tibial artery is raised from the middle and lower regions of the medial aspect of the leg. It has a long pedicle, and it can be used as a free flap to reconstruct the distant soft tissue defects and also as an island flap. We have used this retrograde island flap for surfacing ulcerated areas in six leprosy patients. The flap survived in all cases. At 24 to 60 months follow-up examination, ulceration had not recurred in any of them. The medial knee flap consisting of the skin and subcutaneous tissue of the lower part of the medial side of the thigh and the upper part of the leg, is suitable for covering soft tissue cushion defects of the extremities because of the constant vessels, long pedicle, wide diameter, well-recognizable sensory nerves and less subcutaneous fat. We have used the medial knee flap for the resurfacing sizeable raw areas due to ulceration in three leprosy patients. The flap survived in all cases and there was no recurrence of ulceration during the 70-148 months follow-up period. PMID- 10804973 TI - Drug resistance in leprosy: lessons from past and future perspective. AB - Some recent studies indicate that the problem of drug resistance in leprosy is very much there but the exact picture is not clear. In the emerging scenario with increasing number of new cases with low bacterial load, the conventional in-vivo and most of current in-vitro methods for determination of drug resistance may not help. It is pointed out that newer molecular approaches may be more useful and that it will be important to undertake studies to develop such tools. PMID- 10804974 TI - Community participation in case detection of leprosy in Nagpur district of Maharashtra. AB - Involving special community groups for new case detection is of great importance for achieving the target of elimination of leprosy. During 1998-99, thirty village level Mahila mandals (women's groups), 6950 teachers and students and 34,548 heads of families were co-opted to participate in case detection. They examined 56,113 persons including 378,959 school students and 184,940 family members. Of the examined population, 275 were suspected to be cases of leprosy by mahila mandals, 411 by teachers and students and 747 by heads of families. Subsequent examination of the suspected cases by trained medical officers and paramedical workers confirmed 203 of them to be cases of leprosy. This exercise showed that when proper attempts were made to involve the community, case detection activity became easier, besides helping to disseminate knowledge about leprosy in the community. PMID- 10804975 TI - "Outside project area" leprosy cases in 'A' ward of Mumbai. AB - Analysis of newly registered smear-positive cases in a ward of the metropolitan city of Mumbai, which has a railway terminus during 1990-97 revealed that 72% of the patients came from outside the project area, most of them arising from the States of Uttar Pradesh, Bihar and Orissa. They had unstable and temporary residences in the area and were employed in low income hard labour jobs. Nevertheless, it was found that their treatment completion rate was high. Using different approaches, e.g. through the community leaders of footpath dwellers and railway platform dwellers, and those of different state language groups' colonies, the new entrants were examined periodically and simultaneously proper rapport was maintained with the medical practitioners of the ward for more referrals to leprosy clinic. Such special approach may have to be developed to tackle such situation in other metropolis in the country. PMID- 10804976 TI - Comparative leprosy vaccine trial in South India. PMID- 10804977 TI - Survey of labour in "Usana" rice factory, in Bhandara district (Maharashtra) PMID- 10804978 TI - Involvement of non-allopathy medical students in LEC's. PMID- 10804979 TI - Photo survey for leprosy in Mumbai. PMID- 10804980 TI - Voluntarily reporting leprosy cases in rural area. PMID- 10804981 TI - Report of a special meeting of the Leprosy Elimination Advisory Group (LEAG) with potential partners. Geneva, 12 and 13 April 1999. PMID- 10804982 TI - Guidelines for implementing a disability prevention programme in the field. National Leprosy Elimination Programme (NLEP). PMID- 10804984 TI - An indirect method of estimating VO2 from work load using arm-cranking in adult hemiplegic patients. AB - The purpose of this study was to develop an oxygen consumption (VO2) measurement for adult hemiplegic patients during an incremental exercise arm-cranking test. Fifty-five adult hemiplegic patients performed a three-stage arm-cranking exercise test while VO2 was measured. The control group consisted of thirteen healthy elderly persons who performed a similar arm-cranking test until exhaustion. The results indicated that there was a significant difference in VO2 during rest periods between the adult hemiplegic and control groups. Based on data analysis, we developed an equation to estimate the VO2 for adult hemiplegic patients: VO2(ml/min) = (kgm/min x 4.2 ml/kgm) + ((4.2 ml/kg/min x kg body weight) + 105.6 ml/min) (r = 0.847), while the equation for the controls was VO2(ml/min) = (kgm/min x 4.3 ml/kgm) + ((2.9 ml/kg/min x kg body weight) + 105.5 ml/min) (r = 0.932). There was no significant difference between the estimated VO2 for the new equations (nVO2) and the measured VO2 (mVO2) in the two groups at any stage. In our data analysis, based on an estimated VO2 from the American College of Sports Medicine (ACSM) equation (aVO2), the nVO2 and mVO2 at each stage showed lower aVO2 than either nVO2 or mVO2 in each subject (p < 0.05). These findings suggest that the ACSM equation is unlikely to be applicable for estimating VO2 for one hand arm-cranking exercise. In conclusion, the new equations established in this study, when used with a specific prescribed exercise testing protocol, would allow us to estimate VO2 more accurately for both adult hemiplegic patients and healthy elderly persons. PMID- 10804983 TI - Pranlukast, a cysteinyl leukotriene antagonist, reduces serum eosinophil cationic protein levels in patients with asthma. AB - Cysteinyl leukotrienes (cysLTs) are considered to be important mediators involved in bronchial asthma and the ensuing eosinophilic inflammation. We evaluated the effects of pranlukast, a potent and selective cysLT receptor antagonist, on the clinical course and serum eosinophil cationic protein (ECP) levels of 10 asthmatic patients. A four-week administration of pranlukast increased the morning peak expiratory flow (PEF) (p = 0.007) and decreased as-needed beta 2 agonist use (p = 0.021). Changes in the morning PEF inversely correlated with those in the serum ECP levels (r = -0.80, p = 0.0057). These results suggest that cysLTs are important mediators involved in eosinophilic inflammation, a major pathophysiologic feature of bronchial asthma in humans. PMID- 10804985 TI - Effects of pentoxifylline on sperm motion characteristics in normozoospermic men defined by a computer-aided sperm analysis. AB - This study was conducted to determine whether pentoxifylline has an in vitro effect on sperm motion characteristics in normozoospermic men. The subjects of the study were 15 male volunteers. After the spermatozoa were washed with mBWW medium, the sperm suspension was divided into two aliquots: one was treated with 1 mg/ml pentoxifylline, the other was used as a control. The sperm motion characteristics were examined by an HT-M2030 at 30, 60, 120, 180, 240 and 300 min during continuous exposure to the drug. As results, pentoxifylline increased the curvilinear velocity and the lateral head displacement. However, it did not affect sperm motility, the straight line velocity. Pentoxifylline may improve sperm fertilizing ability by altering the characteristics of sperm motion, not by increasing the number of motile spermatozoa. PMID- 10804986 TI - Malignant paraganglioma arising from the posterior mediastinum: a case report and review of the literature. AB - An unusual case of paraganglioma of posterior mediastinum occurred in a young adult with local recurrence and multiple distant metastasis. Because of its rarity, the determinants of prognosis factor between benign and malignant paraganglioma are uncertain. In this case, we investigated abnormalities of the p53 gene and ras gene mutations in tissues of primary and metastatic lesions. Neither abnormalities of p53 gene nor ras gene mutations were detected. The molecular approach is recommended as a means of clarifying the trend towards the malignancy of paraganglioma. PMID- 10804987 TI - A case of bronchial squamous cell carcinoma in situ detected by sputum cytology. AB - A 64-year-old man underwent a medical checkup in May 1996 and was evaluated as class V using sputum cytology. Chest X-ray examination, bronchoscopy and chest computed tomography (CT) demonstrated no abnormalities. Thereafter, the patient was followed up with chest X-ray, bronchoscopy and chest CT at 3-month intervals. In December 1996, chest CT showed an increased density at the mediastinal side of the left upper bronchus, B1+2. There were no findings on bronchoscopy, but subsequent exfoliative cytology demonstrated keratinized malignant cells in samples obtained from left upper bronchus, B1+2. Although, it was difficult to identify localization of the tumor, left upper lobectomy was performed and the diagnosis of squamous cell carcinoma in situ was finally made. Here, we report on the course of this patient and discuss the diagnostic usefulness of sputum cytology as well as the pathogenesis of lung squamous cell carcinoma. PMID- 10804988 TI - Mechanisms for the biological effectiveness of high-LET radiations. AB - Radiations of high linear energy transfer (LET) have long been known to have greater biological effectiveness per unit dose than those of low LET, for a wide variety of biological effects. However, values of relative biological effectiveness depend considerably on the biological system and in some instances the values are clearly below unity. The differences between high- and low-LET radiations may be due to many factors, almost all of which are related to radiation track structure in one way or another, and some can in principle lead to qualitative as well as quantitative differences between the radiations. Explanations for LET-dependent differences in effectiveness are discussed over a variety of levels from the multicellular and cellular scale down to the DNA scale, with illustrations from radiobiological data. Information from well defined slow light ions provide particularly useful analytic data, but practical issues extend also to neutrons and fast heavy ions, which may compound high- and low-LET features. It is suggested that effectiveness of the radiation is determined predominantly by the complex clustered damage that it produces in DNA, but that for high-LET radiations long-term effects are in some instances limited by single-track-survival probabilities of the traversed cells. PMID- 10804989 TI - Neutron generator (HIRRAC) and dosimetry study. AB - Dosimetry studies have been made for neutrons from a neutron generator at Hiroshima University (HIRRAC) which is designed for radiobiological research. Neutrons in an energy range from 0.07 to 2.7 MeV are available for biological irradiations. The produced neutron energies were measured and evaluated by a 3He gas proportional counter. Energy spread was made certain to be small enough for radiobiological studies. Dose evaluations were performed by two different methods, namely use of tissue equivalent paired ionization chambers and activation of method with indium foils. Moreover, energy deposition spectra in small targets of tissue equivalent materials, so-called lineal energy spectrum, were also measured and are discussed. Specifications for biological irradiation are presented in terms of monoenergetic beam conditions, dose rates and deposited energy spectra. PMID- 10804990 TI - Mutation induction and RBE of low energy neutrons in V79 cells. AB - We have examined the neutron energy dependency of cell killing and mutation induction at the hprt locus in Chinese hamster V79 cells. Monoenergetic neutrons at 0.32, 0.57, and 1.2 MeV were generated at the Hiroshima University Radiobiological Research Accelerator (HIRRAC) Facility, and were used to irradiate cells. The variation in RBE with neutron energy for the end points of cell survival and hprt mutation induction was observed. When compared to 137Cs gamma-rays, all neutron energies were more effective at both cell killing and induction of mutation. Over the range of the neutron energies examined, we found that cytotoxicity increased as the energy decreased from 1.2 to 0.32 MeV. In comparison to gamma-rays, RBEs for cell lethality at 10% survival were 5.7, 6.7, and 7.6 for 1.2, 0.57, and 0.32 MeV, respectively. Mutation induction, on the other hand, was highest at 0.57 MeV with a gradual decrease at 1.2 and 0.32 MeV. RBEs for mutation induction were 9.7, 19.4, and 13.9 for 1.2, 0.57, and 0.32 MeV neutrons. We isolated independent V79 cell mutants at the hprt locus from untreated and neutron-exposed cells and determined the genetic changes underlying the mutation by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. Preliminary results are suggestive of a specific relationship between deletion pattern and neutron energy. PMID- 10804991 TI - Dose estimations of fast neutrons from a nuclear reactor by micronuclear yields in onion seedlings. AB - Irradiations of onion seedlings with fission neutrons from bare, Pb-moderated, and Fe-moderated 252Cf sources induced micronuclei in the root-tip cells at similar rates. The rate per cGy averaged for the three sources, , was 19 times higher than rate induced by 60Co gamma-rays. When neutron doses, Dn, were estimated from frequencies of micronuclei induced in onion seedlings after exposure to neutron-gamma mixed radiation from a 1 W nuclear reactor, using the reciprocal of as conversion factor, resulting Dn values agreed within 10% with doses measured with paired ionizing chambers. This excellent agreement was achieved by the high sensitivity of the onion system to fast neutrons relative to gamma-rays and the high contribution of fast neutrons to the total dose of mixed radiation in the reactor's field. PMID- 10804992 TI - Neutron energy-dependent initial DNA damage and chromosomal exchange. AB - This study was undertaken to investigate the biological effect of monoenergetic neutrons on human lymphocyte DNA and chromosomes. Monoenergetic neutrons of 2.3, 1.0, 0.79, 0.57, 0.37 and 0.186 MeV were generated, and 252Cf neutrons and 60Co gamma-rays were also used for comparison. Biological effect was evaluated two ways. The RBE values with the comet assay were estimated as 6.3 and 5.4 at 0.37 MeV and 0.57 MeV relative to that of 60Co gamma-rays, and chromosome aberration rates were also observed in these different levels of monoenergetic neutrons. The yield of chromosome aberrations per unit dose was high at lower neutron energies with a gradual decline with 0.186 MeV neutron energy. The RBE was increased to 10.7 at 0.57 MeV from 3.9 at 252Cf neutrons and reached 16.4 as the highest RBE at 0.37 MeV, but the value decreased to 11.2 at 0.186 MeV. The response patterns of initial DNA damage and chromosome exchange were quite similar to that of LET. These results show that the intensity of DNA damage and chromosomal exchange is LET dependent. RBE of low energy neutrons is higher than that of fission neutrons. Low energy neutrons containing Hiroshima atomic bomb radiation may have created a significantly higher incidence of biological effect in atomic bomb survivors. PMID- 10804993 TI - Cell cycle and LET dependence for radiation-induced mutation: a possible mechanism for reversed dose-rate effect. AB - A previous study of the mutagenic action of 252Cf radiation in mouse L5178Y cells showed that the mutation frequency was higher when the dose was chronic rather than acute, which was in sharp contrast to the effects reported for gamma-rays (Nakamura and Sawada, 1988). A subsequent study using synchronized cells revealed that the cells at the G2/M stage were uniquely sensitive to mutation induction by 252Cf radiation but not to gamma-rays (Tauchi et al., 1993). A long phase cell population was first subjected to conditioning gamma or 252Cf radiation doses at different dose-rates. The cell cycle distribution of these cells was then observed, and they were then exposed to 252Cf radiation, and the mutation rate was determined. The G2/M fraction increased by 3- to 4-fold when the conditioning doses (2 Gy of gamma or 1 Gy of 252Cf radiation) were delivered chronically over 10 h, but only slightly when the same doses were delivered over a 1 h period or less. Subsequent 252Cf irradiation gave higher mutation frequencies in the cells pre-irradiated with gamma-rays over a protracted period of time than in those exposed with the higher dose-rate gamma-rays. These results suggest that the radiation-induced G2 block could be at least partly (but not totally) responsible for this reverse dose-rate effect (Tauchi et al. 1996). Possible factors which cause the hyper-sensitivity of G2/M cells to mutation induction by neutrons will be discussed. PMID- 10804994 TI - Neutron-energy-dependent cell survival and oncogenic transformation. AB - Both cell lethality and neoplastic transformation were assessed for C3H10T1/2 cells exposed to neutrons with energies from 0.040 to 13.7 MeV. Monoenergetic neutrons with energies from 0.23 to 13.7 MeV and two neutron energy spectra with average energies of 0.040 and 0.070 MeV were produced with a Van de Graaff accelerator at the Radiological Research Accelerator Facility (RARAF) in the Center for Radiological Research of Columbia University. For determination of relative biological effectiveness (RBE), cells were exposed to 250 kVp X rays. With exposures to 250 kVp X rays, both cell survival and radiation-induced oncogenic transformation were curvilinear. Irradiation of cells with neutrons at all energies resulted in linear responses as a function of dose for both biological endpoints. Results indicate a complex relationship between RBEm and neutron energy. For both survival and transformation, RBEm was greatest for cells exposed to 0.35 MeV neutrons. RBEm was significantly less at energies above or below 0.35 MeV. These results are consistent with microdosimetric expectation. These results are also compatible with current assessments of neutron radiation weighting factors for radiation protection purposes. Based on calculations of dose-averaged LET, 0.35 MeV neutrons have the greatest LET and therefore would be expected to be more biologically effective than neutrons of greater or lesser energies. PMID- 10804995 TI - Repair of cellular damage after high LET irradiation. PMID- 10804996 TI - Radiobiological studies using synchrotron-produced ultrasoft X-rays. AB - Ultrasoft X-rays have been extensively used to explore radiobiological mechanisms surrounding cell killing. These studies for the most part have been linked to a small number of X-ray energies. Recently, this field of study has been broadened by the availability of synchrotron-produced ultrasoft X-rays which can be produced at any desired energy. We have taken advantage of the University of Wisconsin Synchrotron to reexamine two fundamental radiobiological questions: Dose RBE vary with different ultrasoft X-ray energies? Dose the fraction of the nuclear volume exposed to equal total X-ray energy modify cell cytotoxicity? The first study focuses on the survival of Chinese hamster V79 and mouse C3H10T1/2 cells irradiated with synchrotron-produced 273 eV and 860 eV ultrasoft X-rays. These two energies, which are available by multilayer monochromatization of the synchrotron output spectrum, exhibit equal attenuation within living cells. Such an isoattenuating energy pair allows the direct examination of how biological effectiveness varies with the energy of the ultrasoft X-rays. In comparing survival results, we find similar biological effectiveness of these two energies for both the C3H10T1/2 and the V79 cells. These results are no consistent with previous findings of increasing RBE with decreasing ultrasoft X-ray energies. In addition, after correcting for mean nuclear based on measurements of cell thickness obtained with confocal microscopy, we find no significant differences in survival between the two ultrasoft X-ray energies and 250 kVp X-rays. These results suggest that RBE does not increase with decreasing energy of ultrasoft X ray between 860 eV and 273 eV. In a second study we introduced an method which allows partial-volume irradiation of live cells using synchrotron-produced ultrasoft X-rays and micro-fabricated irradiation masks. The masks were made by X ray lithography at the University of Wisconsin Synchrotron Radiation Center, and they consist of 1.85-micron-wide stripes of gold 1.35 microns apart plated onto thin silicon nitrate membranes. When placed adjacent to mylar on which live cells are plated, these masks allow cells to be irradiated in a striped pattern with dimensions much smaller than the cell nuclei. Using 1340 eV synchrotron-produced X-rays, we compare the survival of cells subjected to uniform irradiation and cells subjected to partial-volume irradiation. Our results show that, at equal mean dose to the nucleus (i.e. equal total energies deposited), survival is not statistically different for the two treatments over a wide range of doses. Thus, imparting equal energies to smaller intranuclear volumes does not appear to modulate cell killing. PMID- 10804997 TI - High-LET radiotherapy and experiences in improving rural health care in India. AB - A brief back ground of high-LET radiotherapy developments and its current status will be presented. In addition, based on my experience in developing a program on cancer prevention and early detection in rural India, the gulf between sophisticated developments in cancer diagnosis and treatment and the basic needs in the developing world will be presented with some possible solutions. PMID- 10804998 TI - RBE-LET relationships in mutagenesis by ionizing radiation. AB - The paper considers the relationship between the quality of radiation and biological lesions produced by ionizing radiation. The paper provides a brief review of the modelling of induction of strand breakage, chromosome aberration, revertant mutation in bacteria and Drosophila melanogaster. Experimental data are presented for the relative biological effectiveness of helium ions and alpha particles for mutation induction and genome lethality in Escherichia coli. The paper examines the relationship between the mutational events and LET. The RBE LET values for T4 phage, E. coli WP2 and mwh (multiple wing hair) show dependency on LET while the wi (white-ivory) allele mutants show no dependency. PMID- 10804999 TI - RBE-LET relationships of high-LET radiations in Drosophila mutations. AB - The relative biological effectiveness (RBE) of 252Cf neutrons and synchrotron generated high-energy charged particles for mutation induction was evaluated as a function of linear energy transfer (LET), using the loss of heterozygosity for wing-hair mutations and the reversion of the mutant white-ivory eye-color in Drosophila melanogaster. Loss of heterozygosity for wing-hair mutations results predominantly from mitotic crossing over induced in wing anlage cells of larvae, while the reverse mutation of eye-color is due to an intragenic structural change (2.96 kb-DNA excision) in the white locus on the X-chromosome. The measurements were performed in a combined mutation assay system so that induced mutant wing hair clones as well as revertant eye-color clone can be detected simultaneously in the same individual. Larvae were irradiated at the age of 3 days post oviposition with 252Cf neutrons, carbon beam or neon beam. For the neutron irradiation, the RBE values for wing-hair mutations were larger than that for eye color mutation by about 7 fold. The RBE of carbon ions for producing the wing hair mutations increased with increase in LET. The estimated RBE values were found to be in the range 2 to 6.5 for the wing-hair. For neon beam irradiation, the RBE values for wing-hair mutations peak near 150 keV/micron and decrease with further increase in LET. On the other hand, the RBE values for the induction of the eye-color mutation are nearly unity in 252Cf neutrons and both ions throughout the LET range irradiated. We discuss the relationships between the initial DNA damage and LET in considering the mechanism of somatic mutation induction. PMID- 10805000 TI - Neutron carcinogenesis: past, present, and future. AB - An interest in the possible cancer causing ability of neutrons began soon after their discovery. Early use of neutrons from radioactive sources and from cyclotrons led to a need to define risk for such exposures. This need was soon followed by a more tangible need to define risk to the general population of high LET radiation from nuclear fall out and use of the Atomic bomb and possible use of the H-bomb. Neutrons were soon found to be very effective cell killing agents compared to conventional ionizing radiation. However High LET radiation sources and neutrons in particular, come in many different energies and from many types of sources. I will survey the differences between different energy neutrons and conventional types of radiation, particularly with respect to the dose rate of exposures and the influence of repair or lack thereof and more recently the effect of cell cycle distribution on the carcinogenic outcome. I will illustrate these ideas with examples of carcinogenicity studies and mutation studies from my own laboratory and in some cases from the work of others. Lastly I will introduce some possible avenues for molecular studies of neutron effects that might answer such vexing questions as the real risk at very low doses, is repair error free or error prone, do neutrons cause genetic instability for many cell generations after exposure, and others? There remain many questions about the biology of neutron action that require answers if we are to protect the ever increasing number of people exposed to them because of their growing use in medicine, in the military and in commercial industry. PMID- 10805001 TI - Kinetics of mammary clonogenic cells and rat mammary cancer induction by X-rays or fission neutrons. AB - Following the hormonal treatment of rats with high prolactin levels and glucocorticoid deficiency (Prl+/Glc-) for 48 days (Day +48), total recoverable mammary DNA was increased by more than sevenfold, tritiated thymidine uptake by nearly fourfold, and total mammary clonogens by about fivefold. Irradiation with 4, 40, and 80 cGy X-rays on Day +48 increased total mammary carcinomas per rat day-at-risk linearly with dose, and 40 and 80 cGy significantly decreased first carcinoma latency. A dose of 40 cGy X-rays before hormone treatment (Day -1) yielded tumor latencies and frequencies insignificantly different from unirradiated controls but significantly different from those when the dose was given on Day +48. Total carcinomas per rat-day-at-risk were fitted better by a function of dose to the power 0.4 than by a linear function after exposure to 1, 10. and 20 cGy fission neutrons, and 10 and 20 cGy significantly shortened the time to appearance of the first cancer. In contrast to results with X-rays, 10 cGy neutrons on Day -1 yielded tumor frequencies and latencies insignificantly different from 10 cGy neutrons on Day +48. The carcinogenic action of X-rays, but not of neutrons, was thus influenced by total clonogen numbers and/or proliferation rates. PMID- 10805002 TI - Uncertainties of DS86 and prospects for residual radioactivity measurement. AB - Residual radioactivity data of 152Eu, 60Co and 36Cl have been accumulated and it has been revealed in the thermal neutron region that a systematic discrepancy exists between the measured data and activation calculation based on the DS86 neutrons in Hiroshima. Recently 63Ni produced in copper samples by the fast neutron reaction 63Cu(n,p)63Ni has been of interest for evaluation of fast neutrons. Reevaluation of atomic-bomb neutrons and prospects based on residual activity measurements have been discussed. PMID- 10805003 TI - A crack model of the Hiroshima atomic bomb: explanation of the contradiction of "Dosimetry system 1986". AB - There has been a large discrepancy between the Dosimetry system 1986 (DS86) and measured data, some of which data in Hiroshima at about 1.5 km ground distance from the hypocenter are about 10 times larger than the calculation. Therefore its causes have long been discussed, since it will change the estimated radiation risks obtained based on the Hiroshima and Nagasaki data. In this study the contradiction was explained by a bare-fission-neutron leakage model through a crack formed at the time of neutron emission. According to the present calculation, the crack has a 3 cm parallel spacing, which is symmetric with respect to the polar axis from the hypocenter to the epicenter of the atomic bomb. We made also an asymmetric opening closing 3/4 of this symmetric geometry, because there are some data which shows asymmetry. In addition, the height of the neutron emission point was elevated 90 m. By using the asymmetric calculation, especially for long distant data located more than 1 km, it was verified that all of the activity data induced by thermal and fast neutrons, were simultaneously explained within the data scattering. The neutron kerma at a typical 1.5 km ground distance increases 3 and 8 times more than DS86 based on the symmetric and asymmetric model, respectively. PMID- 10805004 TI - Biological dosimetry of atomic bomb survivors exposed within 500 meters from the hypocenter and the health consequences. AB - Seventy-eight atomic bomb survivors were examined for biological dosimetry using chromosome abnormality. They had been exposed within 500 meters from the hypocenter in heavily shielded conditions and were found from NHK-RIRBM joint study carried out from 1966 to 1971. Estimation of the exposure doses for these survivors was made under the following steps; 1) calculation by DS86 system (physically estimated doses) in survivors who had been exposed within 1,500 meters and had precise records of exposure conditions. RBE for the neutron was defined as 10. 2) setting of exposure dose-chromosome aberration curve, and 3) observation of chromosome aberrations in the proximally exposed survivors, for whom biological doses were estimated. Estimation of the exposure doses were possible from the aberration rate of chromosome in the peripheral lymphocytes, even 25-40 years after the exposure. Of the 78 survivors, 96% were estimated to have exposed more than one Sv. Detection of transforming gene(s) of N and K RAS genes in DNAs from non-leukemic survivors was carried out as one of the biological investigations for these heavily exposed survivors. All four survivors examined showed N or K RAS gene mutation. Three of the four healthy survivors had cancer or leukemia 7-10 years after the examination. Further continuous follow-up study of these heavily exposed people will give us more information on the late effects of A-bomb radiation, which may arise in the future. PMID- 10805005 TI - F-value as a chromosomal fingerprint of the quality of radiation. AB - Since the first proposal by Brenner and Sachs (Radiation Res. 140, 134-142, 1994), the F-value, the ratio of inter- to intra-chromosomal interchanges, as a biomarker for the quality of radiation has been a matter of repeated discussion. Controversies seem to stem from the selection of data which are heterogeneous in terms of chromosome scoring criteria and dose range. In the context of the critical evaluation of the validity of the F-value, the cytogenetic data obtained in our laboratory from the in vitro irradiation of human peripheral blood lymphocytes have been re-assessed for the F-value. The results were consistent with the original contention that the densely ionizing radiations showed lower F value. The differential F-value was more pronounced in the low-dose range and disappeared with the increase of the dose, or more precisely with the number of charged particles passing through the cell nucleus. The range of charged particles also plays a role, which makes the F-value of neutrons insensitive to their energy due to a wide variation of the kinetic energy of recoil protons. PMID- 10805006 TI - Health informatics: challenges to progress. PMID- 10805007 TI - Scalable software architectures for decision support. AB - Interest in decision-support programs for clinical medicine soared in the 1970s. Since that time, workers in medical informatics have been particularly attracted to rule-based systems as a means of providing clinical decision support. Although developers have built many successful applications using production rules, they also have discovered that creation and maintenance of large rule bases is quite problematic. In the 1980s, several groups of investigators began to explore alternative programming abstractions that can be used to build decision-support systems. As a result, the notions of "generic tasks" and of reusable problem solving methods became extremely influential. By the 1990s, academic centers were experimenting with architectures for intelligent systems based on two classes of reusable components: (1) problem-solving methods--domain-independent algorithms for automating stereotypical tasks--and (2) domain ontologies that captured the essential concepts (and relationships among those concepts) in particular application areas. This paper highlights how developers can construct large, maintainable decision-support systems using these kinds of building blocks. The creation of domain ontologies and problem-solving methods is the fundamental end product of basic research in medical informatics. Consequently, these concepts need more attention by our scientific community. PMID- 10805008 TI - Clinical terminology: why is it so hard? AB - Despite years of work, no re-usable clinical terminology has yet been demonstrated in widespread use. This paper puts forward ten reasons why developing such terminologies is hard. All stem from underestimating the change entailed in using terminology in software for 'patient centred' systems rather than for its traditional functions of statistical and financial reporting. Firstly, the increase in scale and complexity are enormous. Secondly, the resulting scale exceeds what can be managed manually with the rigour required by software, but building appropriate rigorous representations on the necessary scale is, in itself, a hard problem. Thirdly, 'clinical pragmatics'--practical data entry, presentation and retrieval for clinical tasks--must be taken into account, so that the intrinsic differences between the needs of users and the needs of software are addressed. This implies that validation of clinical terminologies must include validation in use as implemented in software. PMID- 10805009 TI - Medical narratives and patient analogs: the ethical implications of electronic patient records. AB - An electronic patient record consists of electronically stored data about a specific patient. It therefore constitutes a data-space. The data may be combined into a patient profile which is relative to a particular specialty as well as phenomenologically unique to the specific professional who constructs the profile. Further, a diagnosis may be interpreted as a path taken by a health care professional with a certain specialty through the data-space relative to the patient profile constructed by that professional. This way of looking at electronic patient records entails certain ethical implications about privacy and accessibility. However, it also permits the construction of artificial intelligence and competence algorithms for health care professionals relative to their specialties. PMID- 10805010 TI - An approach to policy analysis and development of medical informatics. AB - There are three grand challenges for medical informatics policy: (1) What is it? (2) What should it be? (3) How can we influence its development? To address these challenges requires: (1) an historical analysis of medical informatics policies in a representative sample of countries. This should include an account of major events, the roles of technology, individuals, culture and social settings. Pioneers have been led by visions of what medical informatics should achieve. The role of these visions and the reactions to unmet expectations thus also need to be analysed; (2) a generally applicable medical informatics policy that places the needs of its stakeholders and clients first. Top priorities are to support quality health care delivery and quality management of health care facilities; (3) an explanation of how policies in medical informatics are created and implemented together with a strategy to guide medical informatics professionals in their lobbying efforts. PMID- 10805011 TI - Effective multidisciplinary communication in healthcare: cooperative documentation systems. AB - To support multidisciplinary communication and cooperation in healthcare, cooperative documentation systems (CDSs) have been developed. However, in the current generation of systems, communication problems that can pose a serious threat to smooth and effective cooperation have occurred and remain to be addressed. This paper presents a set of features that need to be considered in the design of a new generation of CDSs to avoid breakdowns in communication and cooperation. Our approach is solidly grounded in linguistic and social theories and based on empirical investigations of communication patterns in multidisciplinary healthcare. It is argued that this work provides a theoretically rigorous approach to the design of CDSs that will enable effective multidisciplinary communication and cooperation in healthcare. PMID- 10805012 TI - Health informatics and community health: support for patients as collaborators in care. AB - Health informatics has much to offer community health care. Computer networks and telecommunications provide particular support that can enhance the collaboration among clinicians, care providers and patients. Special-purpose computer tools referred to as Consumer Health Informatics (CHI) represent the application of computer and information technologies specifically to support the health information and communication needs of patients and lay persons. Research projects like ComputerLink and CHESS demonstrate that CHI is acceptable to patients and promotes self-care and disease management. Three grand challenges must be faced to insure realization of the promise of health informatics to community health care: development of knowledge management and information discovery tools for patients, insurance of health information literacy for all persons, and re-engineering clinical practice to capitalize on patients as full partners in health care. PMID- 10805013 TI - Clinical judgment revisited. AB - It is widely recognised that accessing and processing medical information in libraries and patient records is a burden beyond the capacities of the physician's unaided mind in the conditions of medical practice. Physicians are quite capable of tremendous intellectual feats but cannot possibly do it all. The way ahead requires the development of a framework in which the brilliant pieces of understanding are routinely assembled into a working unit of social machinery that is coherent and as error free as possible--a challenge in which we ourselves are among the working parts to be organized and brought under control. Such a framework of intellectual rigor and discipline in the practice of medicine can only be achieved if knowledge is embedded in tools; the system requiring the routine use of those tools in all decision making by both providers and patients. PMID- 10805014 TI - Electronic patient records in medical practice: a multidisciplinary endeavor. PMID- 10805015 TI - Consultation of medical narratives in the electronic medical record. AB - This article presents an overview of a research project concerning the consultation of medical narratives in the electronic medical record (EMR). It describes an analysis of user needs, the design and implementation of a prototype EMR system, and the evaluation of the ease of consultation of medical narratives when using this system. In a questionnaire survey, 85 hospital physicians judged the quality of their paper-based medical record with respect to data entry, information retrieval and some other aspects. Participants were more positive about the paper medical record than the literature suggests. They wished to maintain the flexibility of data entry but indicated the need to improve the retrieval of information. A prototype EMR system was developed to facilitate the consultation of medical narratives. These parts were divided into labeled segments that could be arranged source-oriented and problem-oriented. This system was used to evaluate the ease of information retrieval of 24 internists and 12 residents at a teaching hospital when using free-text medical narratives divided at different levels of detail. They solved, without time pressure, some predefined problems concerning three voluminous, inpatient case records. The participants were randomly allocated to a sequence that was balanced by patient case and learning effect. The division of medical narratives affected speed, but not completeness of information retrieval. Progress notes divided into problem related segments could be consulted 22% faster than when undivided. Medical history and physical examination divided into segments at organ-system level could be consulted 13% faster than when divided into separate questions and observations. These differences were statistically significant. The fastest divisions were also appreciated as the best combination of easy searching and best insight in the patient case. The results of our evaluation study suggest a trade-off between searching and reading: too much detailed segments will delay the consultation of medical narratives. Validation of the results in daily practice is recommended. PMID- 10805016 TI - An inventory of publications on electronic patient records. AB - This study describes an analysis of 1832 papers dealing with electronic patient records, and indexed in NLM's MEDLINE. Of each retrieved publication the country of origin and the journal in which it was published was determined. Furthermore, insight into the subjects of the publication was obtained by analysing the MeSH terms by which it was indexed. Since 1990 the number of publications on electronic patient records has increased. Publications originated from 43 different countries representing all continents. However, 75% stemmed from only 4 countries. The publications appeared in 379 different journals, of which 26 journals had 10 or more publications. Of all publications, 5.3% had appeared in journals with an impact factor of at least 4.5. The topics most often dealt with were: Hospital Information Systems, Computer Communication Networks, User Computer Interface, Confidentiality and Computer Security. No obvious trends, other than an increased interest in confidentiality and computer security, were observed. PMID- 10805017 TI - Confidentiality issues within a clinical information system: moving from data driven to event-driven design. AB - Within a hospital, the moving of medical information systems from retrospective data-gathering methods to prospective clinical information systems raises the question of the confidentiality of patient data. A method of improving the traditional matrix model usually used to achieve access controls is described. The event-driven model refers to the way a security system ensures that a given user has a valid "need-to" relationship to a given patient. Events are defined as the occurrence of specific data that trigger the creation or the updating of the relationship between the identity of a user and the identity of a patient (e.g., admission, discharge, transfer, prescription, and report). The creation and the deletion of the relationships between users and patients are based on numerous repositories and working lists of patients. This implementation requires an organization of the hospital activities which is able to manage, in a real-time manner, those repositories as closely as possible to the steps occurring during the patient's care process. Although this approach seems to reasonably fit the dynamic of the care process, it adds significant organizational constraints. PMID- 10805018 TI - Automated semantic indexing of imaging reports to support retrieval of medical images in the multimedia electronic medical record. AB - This paper describes preliminary work evaluating automated semantic indexing of radiology imaging reports to represent images stored in the Image Engine multimedia medical record system at the University of Pittsburgh Medical Center. The authors used the SAPHIRE indexing system to automatically identify important biomedical concepts within radiology reports and represent these concepts with terms from the 1998 edition of the U.S. National Library of Medicine's Unified Medical Language System (UMLS) Metathesaurus. This automated UMLS indexing was then compared with manual UMLS indexing of the same reports. Human indexing identified appropriate UMLS Metathesaurus descriptors for 81% of the important biomedical concepts contained in the report set. SAPHIRE automatically identified UMLS Metathesaurus descriptors for 64% of the important biomedical concepts contained in the report set. The overall conclusions of this pilot study were that the UMLS metathesaurus provided adequate coverage of the majority of the important concepts contained within the radiology report test set and that SAPHIRE could automatically identify and translate almost two thirds of these concepts into appropriate UMLS descriptors. Further work is required to improve both the recall and precision of this automated concept extraction process. PMID- 10805019 TI - Online medical records: a decade of experience. AB - The electronic patient record at the Beth Israel Deaconess Medical Center has fundamentally changed the practice of medicine in ways that its developers never foresaw. This type of highly interactive and work flow enabled program is creating new collaborative roles for computers in complex organizations [4]. With the system able to supervise and monitor care, computers are able to perform many care coordination and documentation functions, freeing people to concentrate more on interpersonal interactions and provision of health care services. One of the challenges in the design of electronic patient records to assist health care providers is how to support collaboration while not requiring that people meet face-to-face. Moreover, a greater challenge for each of us as clinicians is to use this technology as a bridge (rather than a barrier) towards better patient doctor relationships. PMID- 10805020 TI - A decision support system for diabetes care: ethical aspects. AB - In this paper the design and implementation of a decision support system for diabetes care is examined from an ethical perspective. It is noted that the system creates potential for enhancing the realization of the principle of autonomy through improved information to patients and participation by patients. However, there is also potential for using the system in a way that is contrary to good health care. It may provide patients with information they are unable to interpret and handle, and it may be used by healthcare authorities for controlling their personnel in ways contrary to good quality working conditions. In order for a decision support system to function as a well-integrated element in ethically based health care, different ethical aspects have to be taken into account during the design of the system. The ethical aspects also constitute one perspective of a systematic re-evaluation and re-design process. PMID- 10805021 TI - The multifarious function of medical records: ethical issues. AB - Medical records comprise history data, physical examination, biosignals, data acquired through in vitro diagnostic tests, images, therapeutic data, and administrative data. Beyond the use of records for therapy, they are also used in research, teaching, the allocation of resources, and for the construction of the patient's personal history. These functions give rise to a number of important ethical issues. The most important ethical issues are the patient's autonomy in decisions pertaining to proposed therapeutic interventions, the confidentiality of the information contained in the records, the reliability of the information for research, and questions related to healthcare policy and the distribution of healthcare resources. PMID- 10805022 TI - Transferring data from one EPR to another: content--syntax--semantic. AB - When data are transmitted between electronic patient record (EPR) systems, we can distinguish several tasks. One task is the definition of structure and semantic content of the data in a message structure. Another task is the mapping of the sending EPR's structure to this message structure. A third task is the mapping of the message structure to the receiving EPR's structure. We describe an approach, which distinguishes clearly between these different tasks and activities. Using this approach we have implemented a data transfer procedure between a cancer registry application and a middleware for healthcare information systems. Our experience showed that the proposed systematic approach helped identify problems for data transfer in an early design phase. It also allowed us to limit modifications of the data exchange procedure to certain tasks or activities when one of the EPR applications was updated. In the end, we could even exchange the underlying message format without having to reimplement the complete interface. PMID- 10805023 TI - Architecture models to facilitate communication of clinical information. AB - This is an account of the development and use of a context model for facilitating the communication of clinical information. Its function is to articulate the principle of context within a reference architecture for the Electronic Health Care Record (EHCR). The work required a re-examination of established models of communication, the purpose being to use them to support an architecture that could be reasonably expected to accommodate future, and by definition unforeseeable, developments in EHCR communication. The Context Model is built upon seven recognized constituents of communication. These constituents, although having their origin in the engineering of signal communication, have been found to be useful for explication both in the verbal and textual communication of narratives between people. The electronic health care record architecture supported by the model is the European prestandard ENV13606-1. PMID- 10805024 TI - ORCA: the versatile CPR. AB - The introduction of computer-based patient records (CPRs) that fully replace paper records proves especially difficult in specialized care, despite the potential advantages of CPRs for patient care and research. Improved data legibility, availability, sharing of records, and decision support may directly benefit patient care. Barriers to the introduction of CPR applications at institutions may be caused by lack of infrastructure, or by financial or organizational issues. To have clinicians interactively enter data at the point of care is still a big challenge. This paper presents an overview of ORCA (Open Record of CAre): a generic CPR, designed for integration with existing systems, presentation of multi-media patient data, and the collection of structured data, directly by clinicians. ORCA can easily be tailored to the needs of a variety of medical specialists without the need for changes to its data model, functionality, or interface. The paper describes the essence of the architecture of ORCA and the user benefits with emphasis on the support of structured data entry. PMID- 10805025 TI - Postmarketing surveillance based on electronic patient records: the IPCI project. AB - Researchers claim that data in electronic patient records can be used for a variety of purposes including individual patient care, management, and resource planning for scientific research. Our objective in the project Integrated Primary Care Information (IPCI) was to assess whether the electronic patient records of Dutch general practitioners contain sufficient data to perform studies in the area of postmarketing surveillance studies. We determined the data requirements for postmarketing surveillance studies, implemented additional software in the electronic patient records of the general practitioner, developed an organization to monitor the use of data, and performed validation studies to test the quality of the data. Analysis of the data requirements showed that additional software had to be installed to collect data that is not recorded in routine practice. To avoid having to obtain informed consent from each enrolled patient, we developed IPCI as a semianonymous system: both patients and participating general practitioners are anonymous for the researchers. Under specific circumstances, the researcher can contact indirectly (through a trusted third party) the physician that made the data available. Only the treating general practitioner is able to decode the identity of his patients. A Board of Supervisors predominantly consisting of participating general practitioners monitors the use of data. Validation studies show the data can be used for postmarketing surveillance. With additional software to collect data not normally recorded in routine practice, data from electronic patient record of general practitioners can be used for postmarketing surveillance. PMID- 10805026 TI - Finding ethical principles and practical guidelines for the controlled flow of patient data. AB - The application of computing to health care, and particularly to electronic patient records, offers major benefits but raises issues of confidentiality and of potential misuse. Sound access control mechanisms are therefore important, but most models focus upon informed consent by the data subject. This raises challenges in mental health care, and for other vulnerable patients including those comatose, and the severely ill and temporarily distressed. Published algorithms which are used to control record access within a controlled environment therefore have value, as a means of ensuring an open and informed, yet ethically sound, solution. The paper describes the background and issues, and gives an example of such an algorithm. PMID- 10805027 TI - Role of the electronic patient record in the development of general practice in The Netherlands. AB - From the end of the seventies practice computer systems have been introduced in Dutch general practice. In 1998 90% of the GPs had a practice computer and 60% used it for medical purposes, with good patient acceptance. Collaboration between pioneer GPs, professional organizations, academic institutions and software manufacturers has been of decisive importance. While the practice computer is early recognized as a powerful tool for administrative purposes, it is increasingly supporting clinical and preventive care, quality of care improvement, clinical training of students and vocational training of general practice trainees. Also for scientific research and practice supporting research, automatization has proven to be innovating. Much attention is paid to designing valid databases and database-related research. In addition, practice automatization has opened new perspectives for health policy and management. Further development of technical and methodological issues is needed, and, very importantly, of patients' and consumers' involvement. EPR technology in general practice does not optimally work without good numerator and denominator data, and a good quality patient-doctor relationship over time. While the EPR is a useful tool to facilitate a better performance, it may enable the doctor to focus more on the health care essentials. PMID- 10805029 TI - Patient information maintained electronically: a new regulatory action in Japan. PMID- 10805028 TI - Design of a decision support system for test ordering in general practice: choices and decisions to make. AB - The increased availability of tests in the past years has been accompanied by an increased number of blood tests ordered by general practitioners. Dutch investigators report a lack of general practitioners' knowledge concerning the indications for blood tests leading to inappropriate and inadequate use of diagnostic tests. Taking advantage of the use of electronic patient records by Dutch general practitioners, the authors replaced the traditional paper forms for test ordering by a decision-support system. The objective of the decision-support system is to change test-ordering behavior. Designing a system to change test ordering behavior, however, required the selection of a method to provide support. To study different methods for changing test-ordering behavior, the authors developed two versions of the decision-support system BloodLink. The first version, BloodLink-Restricted, is based on the notion of restricting the number of choices presented to the general practitioners. The second version, BloodLink-Guideline, is based on the guidelines provided by the Dutch college of general practitioners. PMID- 10805030 TI - Morphological and physiological changes during growth: an update. AB - Skeletal growth and changes in body composition during growth present important variations; body mass index and lean body mass related to age show important gender differences. The process of ossification is developed in two different ways, endochondral and intramembraneous. The former is characterised by the formation of bone from growth cartilage. Intramembraneous ossification is characterised by the formation of bone from a mesenchymal structure, as occurs with the flat bones of the skull. During childhood and adolescence and up to the acquisition of adult stature, two phenomenons are produced simultaneously: the synthesis of new bone from growth cartilage due to the process of endochondral ossification, and modeling-remodeling of previously synthesized bone. Bone growth and mineralisation of its extracellular matrix are simultaneous phenomenons, the final result being the acquisition and maintenance of body bone mass. A positive calcium balance is necessary during adolescence in order to achieve the maximum peak of bone mass and even with the termination of longitudinal growth of bone, the process of mineralisation can last a further 4 years. Childhood and adolescence are the period of life in which the peak of bone mass must be achieved, and if during this time this does not happen there will be a greater risk for the later development of osteoporosis. Regulation of bone mass is a polygenic process and during recent years studies have been centred on the receptor genes of vitamin D and estrogens. A maximum calcium retention during adolescence may influence the achievement of a high peak of bone mass but at a certain level of calcium intake the calcium retention reaches a plateau. The expression of grams of hydroxyapatite per square centimetre has been used clinically, or expressed in volume as g/cm3. From birth until 3 years, the increase represents approximately 30% of the total increase, from 3 years until the beginning of pubertal development the increase is 20%. During pubertal development there is an increase of 30-40% and from the end of growth until the age of 21 years there is an increase of 15-20%. Both prepubertal boys and girls show a progressive increase of leptin levels during the years prior to the onset of puberty and until Tanner's stage 11 and higher levels are observed in girls in this period, possibly in relation to their earlier onset of puberty. This increase of leptin in girls during pubertal development suggests that leptin may be a link between adipose tissue and puberty. PMID- 10805031 TI - Adolescence: macronutrient needs. AB - Dietary needs during adolescence lack specific definitions, and most evidence is derived from indirect indications. The data on dietary needs for energy and proteins are mainly extrapolated from subjects in other age-classes. Lipids and carbohydrates are being progressively considered for preventive purposes since the qualitative distribution of saturated and unsaturated fats and slowly and rapidly absorbed carbohydrates, respectively, seems to be associated with metabolic index predictors of degenerative disorders in later stages of life. The recent results of multicentric autoptic studies in young people from the US indicate that the lipoprotein status of the second decade of life is associated with the first raised arterial lesions in the third decade. The evidence of these links needs further confirmation from ongoing surveys. We must recognize that adolescence is a critical period of life, and food fads may deeply change the dietary habits acquired within the familiar group. As nutritionists, we may suggest that all adolescents should be supplied with nutritional support in terms of education, maybe at school, to improve their knowledge of nutrition. PMID- 10805032 TI - Growth and micronutrient needs of adolescents. AB - OBJECTIVE: This paper focuses on micronutrients in relation to needs throughout adolescence, a period which involved growth and development that occur through a complex interaction of genetic instructions, hormones and environmental influences, many of them of dietary origin. In the context of micronutrient 'needs' it is of special importance to differentiate between the 'nutritional needs' and 'metabolic needs'. Two main questions arise in relation to the micronutrient needs: (1) why are micronutrients necessary? and (2) how are their needs assessed? RESULTS: The 'necessary' amount will differ according to the objectives pursued: (a) to achieve a satisfactory rate of growth and development; and (b) to maintain 'optimal health'. The assessment of micronutrient needs and status has proved to be difficult, but when elucidating and establishing them, it is imperative to arrive at the estimates in the light of their interdependent role in metabolism and functions. The knowledge of micronutrient metabolic needs can be approached through epidemiological observations, bioavailability studies and clinical trials. However, there is a nearly total absence of reports on the particular metabolic and dietary needs of adolescents. CONCLUSION: Thus more studies are required in relation to the effect of features associated with adolescence on 'needs', evaluating their impact on bioavailability and turnover (storage and losses), and the interactions among micronutrients in the assessment of metabolic and nutritional needs. Another aim should be to establish static and functional indexes, reference values and cut-off points in adolescence, to be used in clinical and epidemiological studies. Future studies should focus on needs to determine those required to maintain 'optimal functions' and regarding the potential prevention of chronic adult diseases. PMID- 10805033 TI - Food habits and sport activity during adolescence: differences between athletic and non-athletic teenagers in Switzerland. AB - OBJECTIVE: To describe food habits and dietary intakes of athletic and non athletic adolescents in Switzerland. SETTING: College, high schools and professional centers in the Swiss canton of Vaud. METHOD: A total of 3,540 subjects aged 9-19 y answered a self-reported anonymous questionnaire to assess lifestyles, physical plus sports activity and food habits. Within this sample, a subgroup of 246 subjects aged 11-15 also participated in an in-depth ancillary study including a 3 day dietary record completed by an interview with a dietician. RESULTS: More boys than girls reported engaging in regular sports activities (P<0.001). Adolescent food habits are quite traditional: up to 15 y, most of the respondents have a breakfast and eat at least two hot meals a day, the percentages decreasing thereafter. Snacking is widespread among adolescents (60-80% in the morning, 80-90% in the afternoon). Food habits among athletic adolescents are healthier and also are perceived as such in a higher proportion. Among athletic adolescents, consumption frequency is higher for dairy products and ready to eat (RTE) cereals, for fruit, fruit juices and salad (P<0.05 at least). Thus the athletic adolescent's food brings more micronutrients than the diet of their non-athletic counterparts. Within the subgroup (ancillary study), mean energy intake corresponds to requirements for age/gender group. CONCLUSIONS: Athletic adolescents display healthier food habits than non-athletic adolescents: this result supports the idea that healthy behavior tends to cluster and suggests that prevention programs among this age group should target simultaneously both sports activity and food habits. PMID- 10805034 TI - Dietary habits and nutritional status in adolescents over Europe. An overview of current studies in the Nordic countries. AB - OBJECTIVE: To give an overview of the dietary habits among adolescents in the Nordic countries and to present results from studies showing the relationship between dietary habits and other lifestyle factors, nutritional status and socio economic conditions. DESIGN: A number of nutritional studies among adolescents performed during recent decades using recalls, dietary records and food frequency questionnaire. SETTING: Denmark, Finland, Norway and Sweden. SUBJECTS: Adolescents aged 13-18 y. RESULTS: Food habits are characterized by an irregular meal pattern; many adolescents skip breakfast and also the school lunch, whereas most of them have dinner. However, snacking and light meals are very common, contributing 25-35% of the daily energy intake. Smoking is linked to their dietary habits as well as socio-economic conditions. Dietary intakes of vitamins and minerals are adequate for normal health and growth. Dietary calcium intake is high, whereas the intake of fibre, vitamin D, zinc and selenium and, in girls, iron is below the Nordic recommendations. Relatively low prevalence figures of iron deficiency were found. Many studies show a decrease over time in physical activity. The time spent in sedentary activities, such as television and video watching and computer games has increased during recent decades. CONCLUSION: Overweight and obesity are becoming more prevalent in all the Nordic countries, even though the prevalence figures are far below those in the USA. On the other hand, dieting girls are common, which might be a factor behind their irregular meal pattern and food choice. In a perspective, overweight and diseases attributable to obesity will be an immense challenge in the coming decades for both the individuals and the society as well. PMID- 10805035 TI - Dietary habits and nutritional status in adolescents over Europe--Southern Europe. AB - A literature review of the data on dietary habits, food consumption and nutritional status of adolescents of Southern European countries was performed. Snacking seems to be less frequent and the frequency of eating out in fast food restaurants is much lower than in the USA and in some Nordic Countries. A nutrient pattern rich in total fat (around 40% of the energy intake), in saturated fatty acids (around 13% of the energy intake) and mainly in monounsaturated fatty acids (17-19% of the energy intake), due to a high consumption of olive oil, is often observed in Spanish and Greek adolescents. In Portugal, the percentage of the energy provided by fat is relatively low (31-33% of the energy intake) and in Italy the situation is intermediate regarding total fat, but the intake of olive oil is also high. This means that two important characteristics of the Mediterranean diet, a low consumption of saturated fatty acids and a high intake of carbohydrates have been lost. Regarding micronutrients, there seems to exist a risk of deficiency, particularly for calcium, iron and zinc. The prevalence of overweight/obesity is high, around 15 25%, being higher than in the Nordic countries but lower than in the USA. There is no clear indication of trends, due to the different methods and criteria used to define obesity. The serum cholesterol levels of adolescents has increased during the last two decades in Spain, Greece and Italy, and nowadays are similar to those in the USA, but still lower than in the Nordic countries. In Portuguese adolescents, the cholesterol levels seem to be lower, but the situation is not well known. PMID- 10805036 TI - Dietary habits and nutritional status in adolescents in Central and Eastern Europe. AB - The overall situation as regards dietary intake and nutritional status in the countries of Central and Eastern Europe has varied considerably during previous decades; however, after the political, social and economic changes during the nineties these differences have increased further, especially in the areas with low GDP and war, and in the regions affected by radionuclides etc. No systematic surveillance systems in representative samples of adolescents were in effect at the beginning of the nineties in Central and Eastern Europe. There exists relatively more data on nutritional status characterized by body mass index (BMI) and skinfold thickness measurements. Food intake has mainly been followed in smaller samples of adolescents, and or in special groups (athletes, obese, vegetarians, allergies, Romanies, pupils of special schools etc.). In the countries and/or areas with sufficient GDP and income the food intake has most often been too abundant when considering the real needs of the growing organism, especially from the point of view of energy output. The composition of the diet is not adequate, i.e. too much protein, fats, sugar etc. This situation is similar to that in Western countries, USA etc. On the other hand, adolescents in Central and Eastern Europe consume too few vegetables, fruit and milk products in their diet (i.e. low intakes of Ca, fibre and vitamin C), which is less apparent in Western countries. This is mainly due to bad eating habits and nutritional traditions in the families, but also due to the changes of the prices of the mentioned foodstuffs after political and economic changes in this part of the world. Certain nutritional deficiencies were also shown as factors increasing the health risks from the Chernobyl disaster, mainly in the Ukraine, Belorussia and certain parts of Poland. PMID- 10805037 TI - Nutritional status and food intake in adolescents living in Western Europe. AB - This review covers surveys of nutritional intake in adolescents living in several countries of Western Europe. Current definitions of nutritional status are discussed, with a special emphasis on adolescent years. It is generally observed that obesity rates are increasing (especially those of massive obesity) in young people, whereas declared energy intakes are decreasing. Average daily energy input seems adequate in adolescents of Western Europe. However, fat (especially saturated) intake is high while that of CHO and fiber is low. Proteins are mainly (two-thirds) from animal sources. Average micronutrient intakes correspond to recommended values in most cases, but there are a few exceptions (calcium and iron) that are low, particularly in girls. Specific problems become frequent at adolescence, such as dieting, smoking, getting low quality foods away from the home, etc. These behaviors may induce adverse nutritional conditions. On average, nutritional problems at adolescence do not appear to be more severe than at other ages, however they may exert a strong deleterious impact on future health. PMID- 10805038 TI - Metabolic adaptations in pregnancy and their implications for the availability of substrates to the fetus. AB - During the first two-thirds of gestation, the mother is in an anabolic condition, increasing her fat depots thanks to both hyperphagia and enhanced lipogenesis. During the last third of gestation, the mother switches to a catabolic condition. Glucose is the most abundant nutrient crossing the placenta, which causes maternal hypoglycemia despite an increase in the gluconeogenetic activity. Adipose tissue lipolytic activity becomes enhanced, increasing plasma levels of FFA and glycerol that reach the liver; consequently there is an enhanced production of triglycerides that return to the circulation in the form of very low density lipoproteins (VLDL). Glycerol is also used as a preferential gluconeogenetic substrate, saving other more essential substrates, like amino acids, for the fetus. Under fasting conditions, fatty acids are converted into ketone bodies throughout the beta-oxidation pathway, and these compounds easily cross the placental barrier and are metabolized by the fetus. An enhanced liver production of VLDL-triglycerides together with a decrease in adipose tissue lipoprotein lipase (LPL) and an increase in plasma activity of cholesterol ester transfer protein causes both an intense increment in these lipoproteins and a proportional enrichment of triglycerides in both low and high density lipoproteins. Maternal triglycerides do not cross the placenta, but the presence of LPL and other lipases allows their hydrolysis, releasing fatty acids to the fetus. Under fasting conditions, the maternal liver uses circulating triglycerides as ketogenic substrates. Around parturition there is an induction of LPL activity in the mammary glands, driving circulating triglycerides to this organ for milk synthesis, allowing essential fatty acids derived from the mother's diet to become available to the suckling newborn. PMID- 10805039 TI - Young people's alcohol consumption from a European perspective: risks and benefits. AB - OBJECTIVE: This paper aims to examine the functions of adolescent drinking by (a) comparing drinking patterns in Mediterranean and Northern European countries, and (b) examination of the beneficial aspects of drinking and pub-going in a Dutch survey. DESIGN: Data from cross-sectional European surveys in 1988 (Eurobarometer) and 1995 (ESPAD-study) and a Dutch cohort study (three waves, respondents were followed from 1989 to 1994) were used. SUBJECTS: The participants in the Eurobarometer-study were 11,729 people from 12 countries who were interviewed at home. The participants in the ESPAD study were 15-16 y olds from 26 European countries interviewed at school. The participants in the Dutch cohort study were 1,063 12-18 y-olds interviewed at school (first wave) and at home (second and third wave). RESULTS AND CONCLUSIONS: The drinking patterns of young people in Mediterranean countries can be characterized as innovative and those of young people in Northern countries as rebellious. The main health risk associated with the innovative pattern is the volume of consumption and associated chronic consequences. The main health risk associated with the rebellious drinking is intoxication and the associated risks concerning violence, traffic accidents and acute health consequences. Furthermore, adolescent drinking and going out should not only be interpreted in terms of risk and negative effects, but seem to have important social and developmental benefits for young people. PMID- 10805040 TI - Obesity in young Europeans: genetic and environmental influences. AB - BACKGROUND: Nutrition, lifestyles and genetics are increasingly involved in the maintenance of health and the prevalence of several non-communicable diseases, such as cardiovascular illnesses, diabetes, cancer etc. In this context, obesity appears as a complex multi-factorial condition resulting from an imbalance between energy intake and expenditure, which has been associated with the genetic background (more than 50 genes have been located in the human gene map associated to obesity), but also with environmental forces such as reduced physical activity (more than 60% of Europeans spend more than 3 h sitting at work) and with the over-consumption of fat-rich and high energy yielding foods (fat oxidation is poorly regulated as compared with other fuel substrates). SUBJECTS: Furthermore, subject selection was quota-controlled to make the sample nationally representative by various socio-demographic factors based on the most recent official statistics (census data) in each member state. Overall, there were 15,239 subjects older than 15 y in the EU (17% of them were younger than 24 y old). RESULTS: The obesity problem appears to be increasing rapidly in children and adolescents as well as in adults with an average rate of obese subjects of about 10% in the European Union, however several factors should be taken into consideration, such as the cut-off references, the age group and the conditions of the data collection when processing the information about obesity prevalence. CONCLUSIONS: Management of the increasing epidemic of obesity in young people must involve prevention strategies concerning nutritional education and physical activity programmes. PMID- 10805041 TI - Eating disorders: a situation of malnutrition with peculiar changes in the immune system. AB - Eating disorders, such as anorexia nervosa and bulimia nervosa, are significant public health concerns for a great deal of the population, and thus are even considered to be epidemics. These syndromes have a common aim: the pursuit of a desirable and extremely low weight, which is obviously very far from the ideal body weight. Therefore, these patients show abnormal food behavior, leading to a situation of malnutrition. Nutrients play an important role in the development and functionality of the immune system. Thus, the assessment of immunological parameters acquires great interest as a useful tool to evaluate the nutritional status of these patients. In addition, it is very well known that a depleted immune system as a consequence of malnutrition is linked to an increased susceptibility to infections. However, an extensive literature has pointed out that anorexic patients, even though severely malnourished, are relatively free from infectious diseases. As the immune system is altered by distorted food behaviors, such as in case of eating disorders, the awareness of the characteristics of other systems involved in these pathologies, and therefore altered, would be very helpful for the understanding of the mechanisms triggered in these syndromes. In fact, the interactions among the immune system and the remaining systems in eating disorders are beginning to be studied. Finally, the main goal is to limit the evolution of these illnesses through an early diagnosis and appropriate therapy to subsequently get a constant and definitive cure for the patients. PMID- 10805042 TI - Body perceptions and slimming attitudes reported by Spanish adolescents. AB - OBJECTIVE: To assess the relationship between body mass index (BMI) and the desire to lose weight, body parts wanted to be reduced and weight-loss practices. DESIGN: A standardized questionnaire was self-completed by the participants. Anthropometric measurements were obtained by a research assistant. SUBJECTS: Forty males and 105 females from 12 to 18 y old randomly selected from education centers in Madrid, Spain. RESULTS: Body dissatisfaction in 56% of the females and the 27% of the males was observed. In spite of the low prevalence of overweight (10% males and 6% females), many adolescents reported themselves to be overweight: more girls (27%) than boys (12.5%) (P < 0.05) by Q2 (6.26) were cognitively inclined to weight loss even though they were not overweight (body mass index, BMI > 25 kg/m2). Adolescents desired a slimming of the abdomen (82% males and 52% females), thighs (18% boys and 39% girls), hips (6% and 18% males and females, respectively) and buttocks (23% males and 42% females). Methods selected to lose weight were: physical activity or sport (73% boys and 68% girls), not to have snacks (66% girls and 23% boys), select foods with low energy content (42% boys and 37% girls), follow own diet (31% girls and 23% boys), follow medical advice (31% boys and 17% girls) and trying to be amused to forget foods (13% girls and 11% boys). PMID- 10805043 TI - Drugs-nutrient interactions: a potential problem during adolescence. AB - The concept of drug-nutrient interactions is not new, but it has only recently gained currency in medicine. Although the elderly are normally considered to be at particular risk, other groups may also be at risk: infants, adolescents, pregnant women, alcohol and tobacco users, etc. In infants and adolescents there are several factors that may influence the possible interactions: firstly, nutrient needs are usually higher, mainly micronutrients; systems for detoxification of anutrients are not complete; the tendency to restricted diets (especially girls) that are unable to cover the actual recommended intakes for a number of micronutrients (i.e. vitamins); and the dangerous increase in alcohol consumption either in males or females. Administration of drugs in population with adequate vitamin intake is usually not a problem, but administration of drugs in those with borderline intake of vitamins or in patients with low nutritional status can result in symptomatic vitamin deficiency states. The groups at risk of poor vitamin status are smokers (a high proportion of adolescents are active smokers); dieters (skipping meals and dieting to lose weight frequently compromise micronutrient intake, and it should be considered that it is extremely difficult to meet all the requirements at intakes of less than 1,200 calories per day), oral contraceptive users, and pregnant and lactating women, excessive alcohol users, etc. The chapter also focuses on the case of folate: rapidly dividing tissues during the adolescent growth spurt increase requirements for folate. Because of this increased need, folate status appears to be of concern during the age of this rapid growth. A variety of drugs are known to interfere with vitamin utilization by blocking or altering transformation of the vitamin to its metabolically active form. Serum folate levels are known to be low in a high percentage of patients with rheumatoid arthritis, suggesting that aspirin alters the transport of folate by competition for binding sites on serum proteins. Methotrexate, a drug commonly used at low doses for the treatment of psoriasis, rheumatoid arthritis and certain liver disorders, limits the availability of methyl groups derived from one-carbon metabolism by inhibiting competitively a key enzyme in the intracellular folate metabolism. In humans, the antiepileptic drug valproic acid (VPA) is associated with two major adverse effects: teratogenicity and folate deficiency. The mechanisms by which VPA exerts the teratogenic or antifolate effect remain unclear, but an alteration in the methionine cycle is the strongest hypothesis proposed. PMID- 10805044 TI - Food allergy and intolerance in children and adolescents, an update. AB - Epidemiological surveys demonstrate that rapid increase in allergic diseases is a real phenomenon. In developed countries they are about the commonest chronic diseases, reaching between 15% and 30% of the population. Adverse reaction to food can be divided into toxic reaction and non-toxic reactions. The non-toxic reactions are divided into non-immune mediated and immune mediated, these are considered food allergic reactions. We showed our experience in a 4 y survey, individualized by food allergens during the first two years of life. In Spain egg white protein is the most common allergen followed by cow's milk and peanuts. These three food items represent half of the sensitizations in children under 2 y of age. After 4 y sensitivities to vegetable allergens such as nuts, fruits and legumes are most frequent. The diagnosis of food allergy is still problematic, even in the case of atopy or IgE mediated hypersensitivity. There is a lack of standardized diagnostic procedures; the only test accepted as 'gold standard' for confirmation of food allergy and in general for food intolerance, is a properly performed double blind placebo-controlled oral food challenge. Negative results should be always followed by an open food challenge. This test should only be conducted in patients with a good medical condition and in a clinic or hospital setting, and only if trained personal and equipment for treating systemic anaphylaxis are present. Contraindications to a challenge test are limited to those situations that can be hazardous for the patient in relationship to the studied food. The treatment of food allergy and intolerance is avoiding the implicated food as long as necessary, until tolerance appears. Prevention of food allergy is the first goal of every pediatric allergologist. Controlled trials of food allergy prevention have been performed only in high allergic risk children. PMID- 10805045 TI - The new British Hypertension Society guidelines: a view from primary care. PMID- 10805046 TI - Ambulatory blood pressure monitoring: which arm? AB - To determine the effects of routinely selecting the nondominant arm for ambulatory blood pressure monitoring (ABPM) on estimates of patients' blood pressure (BP) and to evaluate the practise of using manual BP from one arm and ambulatory BP from the other on the estimation of white coat effect (WCE), an observational study was conducted in 10 volunteers, exhibiting an interarm resting clinic systolic BP (SBP) difference > or =10 mm Hg. The main outcome measures were: (i) average ambulatory SBP measured on right and left arm simultaneously during 24 h, and (ii) estimate of WCE derived, by current practise, as the difference between the referral clinic BP (the higher of the manual readings from both arms) and ambulatory non-dominant arm BP, contrasted with the WCE calculated as the difference between clinic and ambulatory readings from the same arm (the arm with the higher manual readings). The supine referral clinic SBP was 16+/-6 mm Hg higher in the right compared with the left arm. Average 24 h ambulatory SBP was 6+/-7 mm Hg higher in the right arm (range +17 to -3 mm Hg), P = 0.025. Diastolic BP measurements mirrored the systolic findings. One-third of the WCE, estimated by current practise, could be attributed to inconsistency in the choice of arm for BP measurement. Thus, inconsistency in the selection of arms for BP measurement, by different techniques, may confound estimation of patients' cardiovascular morbidity risk. PMID- 10805047 TI - Twenty-four hour blood pressure after exercise in patients with coronary artery disease. AB - The objective was to assess the influence of a cardiac rehabilitation training session on blood pressure measured shortly after exercise and during the subsequent 24 h in patients with stable coronary artery disease. Blood pressure was measured conventionally and by use of 24-h ambulatory blood pressure monitoring in seven men, mean age 53+/-8 (s.d.) years, after participation in a cardiac rehabilitation session and, in randomised order, on a non-exercise control day. Conventional blood pressure averaged 112+/-7/77+/-5 mm Hg in the sitting position on the control day and was not different at the same time of the day shortly after the patients had participated in a cardiac rehabilitation training session. Standing systolic pressure was lower by 7.8+/-4.3 mm Hg (P < 0.005) after exercise compared to the control situation, but this was not associated with orthostatic symptoms. However, ambulatory monitoring showed no differences in blood pressure with the non-exercise day during the subsequent 24 h period. In conclusion, standing but not sitting blood pressure was slightly lower shortly after a cardiac rehabilitation session, but the postexercise orthostatic hypotension was not sustained during normal activities of daily living. PMID- 10805048 TI - Influence of self-measurement of blood pressure on the responder rate in hypertensive patients treated with losartan: results of the SVATCH Study. Standard vs Automatic Treatment Control of COSAAR in Hypertension. AB - Home measurement of blood pressure (BP) can improve compliance. The aim of this study was to evaluate if the efficacy of losartan in hypertension could be enhanced by providing patients with a device for home BP measurement. In this open, randomised, prospective, multi-centre study in 244 Swiss practices, patients with mild-to-moderate hypertension were randomised to a group receiving a home BP measuring device (OMRON) (group 2), or to a group where this device was not provided (group 1). After 8 weeks of treatment with losartan, the responder rates between subjects performing home measurement of BP were compared with those without self-measurement, whereby exclusively sphygmomanometric office BP values were considered. A total of 622 subjects completed the study. Treatment with losartan significantly reduced diastolic (DBP) and systolic BP (SBP) (P < 0.0001). Overall, the group with home BP measurement showed an increased responder rate (DBP < or =90 mm Hg) by 6.4% (59.8% vs 66.2%; 0.05 < P < 0.1). This difference was mainly due to female patients (64.1% vs 73.2%), where it reached statistical significance (P < 0.01). Pretreated patients showed a 9.0% improvement of response in the home measurement group as well (0.05 < P < 0.1). The responder rate in newly treated subjects was relatively high, reaching 79.5% (DBP < or =90 mm Hg or reduction > or =10 mm Hg DBP from baseline). However, home measurement did not significantly improve BP control in these new patients (81.4% vs 77.7%). Overall, home measurement can lead to a slight improvement of BP control. This improvement was most evident in females, reaching significance. PMID- 10805049 TI - Arterial hypotension: prevalence of low blood pressure in the general population using ambulatory blood pressure monitoring. AB - BACKGROUND: Chronic constitutional hypotension has been described in a proportion of the population, and has a symptom complex ascribed to it. The true prevalence of low blood pressure in the normal population has not been defined. AIM OF STUDY: This study was undertaken to determine the prevalence of low blood pressure states, as measured using ambulatory blood pressure monitoring, in a general population cohort, and to determine the association between low blood pressure and clinical and demographic variables. PATIENT POPULATION: The population enrolled were a cohort of mainly urban dwelling Irish subjects, either employees or spouses of employees of a major national bank. METHODS: Subjects had an ambulatory blood pressure monitor fitted between 09.00 and 12.00 and wore the monitor for 24 hours. The subjects also filled out a detailed lifestyle questionnaire, and kept an activity diary. Blood was drawn for serum electrolyte estimation. RESULTS: A total of 254 subjects were included, 49% of whom demonstrated hypotensive events. Hypotensive means and individual hypotensive values were more frequently found in women, and occurred in a group of individuals with a distinct body habitus, specifically thin subjects, with a lower creatinine suggesting a smaller muscle mass. Hypotensive events in these subjects were associated with a low risk cardiovascular profile, in that subjects who displayed these events had a lower blood pressure, a lower weight and were less likely to have a positive family history of hypertension or vascular disease. CONCLUSION: Hypotension is common in the general population and is associated with a distinct body habitus. It carries a generally benign cardiovascular risk factor profile. PMID- 10805050 TI - Hypertension in a black population: prevalence and biosocial determinants of high blood pressure in a group of urban Nigerians. AB - AIMS: To define the prevalence of hypertension, a major cause of morbidity and mortality in blacks, and related biosocial factors in an urban African population group. METHODS: The setting was that of a civil service population in Ibadan, a major city in Southwestern Nigeria. Nine hundred and ninety-eight civil servants selected by multistage sampling participated in the survey. Biosocial data including smoking history, alcohol use and level of physical activity; anthropometry, blood pressure and plasma glucose measurements were obtained. Diagnosis of hypertension was based on blood pressure of > or =160/95 mm Hg or known hypertensive on treatment. RESULTS: The overall prevalence rate of hypertension was 10.3% (CI, 8.4%, 12.2%), rates of 13.9% and 5.3% were obtained in men and women respectively in spite of a much higher rate of generalised obesity in the latter. Hypertension was associated with higher salary grade level, but there was no relationship found with regular exercise, smoking and alcohol. Obesity (body mass index (BMI) > or =30 kg/m2) was associated with hypertension only in women. A two-sided t-test demonstrated age, waist circumference, waist to hip ratio (WHR) and plasma glucose level as significant variables. In multivariate ANOVA models of systolic blood pressures, age, male sex and BMI were highly significant factors (P < 0.0001) and plasma glucose was also significant (P < 0.016); the same variables (except plasma glucose) were associated with diastolic blood pressures. In logistic regression models the variables which predicted hypertension were WHR, plasma glucose, age, sex and family history of diabetes. CONCLUSIONS: Prevalence of hypertension in the study was comparable to recently reported rates in urban Nigeria and similar populations in Africa. The biosocial determinants of hypertension in the urban black population were age, male gender, higher socio-economic status, BMI, plasma glucose, generalised and central adiposity. Regional fat distribution was a stronger predictor of hypertension than generalised obesity in the population. PMID- 10805051 TI - A systematic review of patient information leaflets for hypertension. AB - OBJECTIVE: To review patient information leaflets on hypertension to determine the quality of information currently available to patients. DESIGN AND SETTING: A standardised systematic rating of patient information leaflets for hypertension in the UK. MAIN OUTCOME MEASURE: A quality score per leaflet based on a pre determined rating scale. using recognised criteria with marks allocated for content, writing style, readability and design. RESULTS: Sixty-one leaflets were received, 42 from the Internet and 19 from other sources. Leaflets could achieve a maximum of 84 points. Scores ranged from 30 to 70 for non-Internet leaflets and 28 to 67 for Internet leaflets. Leaflets produced by Boeringer Ingelheim for the British Hypertension Society and Greenlines Publishing (with a medical education grant from Knoll Ltd) scored highest overall. A detailed analysis of the non Internet leaflets showed that few leaflets contained the full range of information considered important though most should be readable by the majority of the adult population and are written using appropriate language and typeface. CONCLUSION: High quality information is available for patients, though some leaflets fall below an adequate standard. Professionals providing advice to patients should have some knowledge of what constitutes good quality information and be critical of the resources they use. Ideally surgeries and clinics should stock a range of the best information available so that patients can chose the leaflet most appropriate to their needs. PMID- 10805052 TI - Comparison of the AT1-receptor blocker, candesartan cilexetil, and the ACE inhibitor, lisinopril, in fixed combination with low dose hydrochlorothiazide in hypertensive patients. AB - AIM: To compare candesartan cilexetil and lisinopril in fixed combination with hydrochlorothiazide with respect to antihypertensive efficacy and tolerability. METHODS: This was a double-blind (double-dummy), randomised, parallel group comparison in patients with a mean sitting diastolic blood pressure 95-115 mm Hg on prior antihypertensive monotherapy. Treatments were candesartan cilexetil/hydrochlorothiazide 8/12.5 mg once daily (n = 237) and lisinopril/hydrochlorothiazide 10/12.5 mg once daily (n = 116) for 26 weeks. The primary efficacy variable was change in trough sitting diastolic blood pressure. RESULTS: Changes in mean sitting diastolic blood pressure did not differ significantly between the groups (mean difference 0.5 mm Hg; 95% confidence interval -1.6, 2.7, P = 0.20). No significant differences between the groups was found for other haemodynamic variables (sitting systolic blood pressure, standing blood pressure, sitting/erect heart rate, and proportion of responders and controlled patients). Both drugs were well tolerated but the proportion of patients with at least one adverse event was significantly greater in the lisinopril group (80% vs 69%, P = 0.020). The proportion of patients spontaneously reporting cough (23.1% vs 4.6%) and discontinuing therapy due to adverse events (12.0% vs 5.9%) was also higher in the lisinopril group compared with the candesartan cilexetil group. CONCLUSIONS: The fixed combinations of candesartan cilexetil and hydrochlorothiazide 8/12.5 mg and lisinopril and hydrochlorothiazide 10/12.5 mg once daily are equally effective as antihypertensive agents. The fixed combination containing candesartan cilexetil is better tolerated than that containing lisinopril. PMID- 10805053 TI - Master--shall I write your article? Pushing the genie back into the bottle. PMID- 10805054 TI - Methodological and political issues in clinical pharmacology research by the year 2000. AB - Parallel groups in a large, multicenter, phase III "pivotal" randomized clinical trial (RCT) with clinically relevant end-points are seen by the medical community as the "gold standard" of clinical research. However, there are limitations, some methodological and others political. The main one is the external validity of the method because a treatment, as studied in RCTs, does not necessarily reflect how it is used in clinical practice. Also, the method, as it stands, is not really predictive of the success in a particular patient of a certain intervention studied in a trial. To overcome these methodological drawbacks, different options have been implemented. The most important ones are: (1) the performance of pragmatic RCTs intended to address effectiveness rather than efficacy; (2) meta analysis; and (3) the use of observational studies, with or without a comparison group. Recent experience has shown that type-A adverse drug reactions (ADRs) related to a specific class of drugs have been successfully characterized throughout cohort- or population-based case-control studies, whereas the evidence linking a specific drug entity to a type-B ADR, apparently severe enough to withdraw the drug from the market, has come mainly from case reports or case series. Other limitations of the RCT are more of a political nature. These large "pivotal" trials are mostly sponsored by the pharmaceutical industry, and to guarantee the scientific and ethical integrity of data produced, they are performed following standard operating procedures (SOPs) and good clinical practice (GCP) guidelines. Sometimes industry is not interested in sponsoring trials; thus, RCTs performed are in practice highly biased because of their potential economical profits. Furthermore, applying SOPs and GCPs is expensive and difficult to implement, and it is hard to find funding in public institutions. As a result, there is an urgent need to create a network of independent, skilled groups interested in sponsoring and performing institutional RCTs following "user friendly" GCP when the profits are low, but scientific interest high. PMID- 10805055 TI - Probucol reduces lysophosphatidylcholines in low-density lipoprotein. AB - OBJECTIVES: Lysophosphatidylcholine (LPC) in low-density lipoprotein (LDL) comes into notice as an important atherogenic substance. METHODS: Since the effect of probucol, an antioxidative lipid-lowering drug, on LPC molecular species has not been elucidated, two LPC molecular species, stearoyl LPC (SLPC) and palmitoyl LPC (PLPC), were measured in LDL using high-pressure liquid chromatography. LDL was obtained from 11 hyperlipidemic patients, including 9 diabetic patients, in comparison with 11 age- and gender-matched controls. RESULTS AND CONCLUSION: Hyperlipidemic patients showed nearly twofold higher levels of SLPC and PLPC per gram of LDL protein than those of controls. All hyperlipidemic patients were treated with oral administration of 500 mg/day of probucol for 3 months. Both LPCs in LDL were significantly reduced to control levels and were increased again up to the pretreatment levels 4 weeks after cessation of the treatment. Therefore, probucol has a potent effect in reducing LPC and may contribute to decreasing the atherogenicity of LDL. PMID- 10805056 TI - Haemodynamic interactions between the novel calcium sensitiser levosimendan and isosorbide-5-mononitrate in healthy subjects. AB - OBJECTIVE: The new calcium sensitiser levosimendan also possesses vasodilatory effects due to potassium-channel opening. The aim of the present study was to assess the possible haemodynamic interactions between levosimendan and isosorbide 5-mononitrate in young healthy men. METHODS: The study was crossover, placebo controlled, double blind, randomised, and it comprised of four study days with one medication--levosimendan, isosorbide-5-mononitrate, levosimendan plus isosorbide-5-mononitrate or placebo--given on each. Levosimendan was administered i.v. as an initial bolus dose of 12 microg/kg over 10 min, followed by a continuous infusion of 0.2 microg/kg/min for a total time of 2 h. Isosorbide-5 mononitrate (20 mg) was given orally as a single dose. Leg blood flow and venous capacity (venous occlusion plethysmography), cardiac output (impedance cardiography), skin blood flow (laser-Doppler flowmetry), blood pressure and heart rate were recorded at baseline, and 20 min, 1 h, 2 h, 4 h and 6 h after the start of the infusion. An orthostatic test was performed at baseline and at 2 h 15 min. Twelve healthy, male subjects were included. Their mean age was 24 years (range 20-34 years). RESULTS: Levosimendan increased leg blood flow by 32%, and no additive effect of the combination of levosimendan and isosorbide-5 mononitrate was observed. The effects of levosimendan on heart rate and blood pressure were minimal and in close conformity with previous studies. In general, there were no additive effects of the combination compared with each drug alone at rest. The only additive effect was seen in the orthostatic response. Heart rate increased by 40 beats min(-1) with the combination (95% confidence interval compared with placebo 11-24 beats min(-1)), by 30 beats min(-1) with levosimendan (2-15 beats min(-1)), by 28 beats min(-1) with isosorbide-5-mononitrate (1-15 beats min(-1)), and by 22 beats min(-1) with placebo. Furthermore, three subjects were unable to stand upright for the stipulated time with the combination, and the orthostatic test had to be discontinued prematurely. There were no changes in the conduction intervals in the electrocardiogram on any of the treatments. The combination had no influence on the occurrence of headache compared with isosorbide-5-mononitrate alone. CONCLUSION: No major additive haemodynamic effects of the combination of levosimendan and isosorbide-5-mononitrate compared with each drug alone could be observed at rest. However, during an orthostatic test, the circulatory response was significantly potentiated with the combination, and three of the subjects were unable to stand upright for the stipulated time. PMID- 10805057 TI - The efficacy of buffered ketoprofen in postoperative pain after third molar surgery. AB - OBJECTIVE: To evaluate in a randomised, double-blind, placebo-controlled trial, the efficacy (time to onset of meaningful pain relief) of single doses of buffered ketoprofen (12.5 mg) and ibuprofen (200 mg) in 180 patients with postoperative pain after third molar surgery. METHODS: 180 adult patients who had undergone third molar surgery under general anaesthesia participated in this study. After dosing, patients recorded their time to meaningful pain relief, pain intensity on both visual analogue scales and verbal rating scales, pain relief and the need for additional analgesia. Pain recordings were made at fixed time points over a 6-h investigation period. RESULT: Buffered ketoprofen (12.5 mg) provided quicker meaningful pain relief than placebo (P = 0.023). For secondary efficacy measures (SPIDS4, SPIDS6, TOTPAR4, TOTPAR6), the buffered ketoprofen was significantly superior to both placebo (P < 0.001) and ibuprofen (200 mg) (P < 0.05). Similarly, the amount of time before taking an escape analgesic was significantly less in the placebo group than both the ibuprofen and buffered ketoprofen groups (P < 0.03). CONCLUSIONS: Buffered ketoprofen (12.5 mg) provides effective pain control in the early postoperative period after third molar surgery. This ketoprofen preparation was also superior to ibuprofen (200 mg) with respect to both reducing pain intensity and providing an earlier onset of pain relief. PMID- 10805058 TI - Structure-effect relationships of amiodarone analogues on the inhibition of thyroxine deiodination. AB - OBJECTIVES: Amiodarone (AMI) has proven to be a potent anti-arrhythmic compound. Due to the structural similarity between AMI and thyroid hormone, it is possible that the drug could inhibit the activity of the 5'-thyroxine-deiodinase. METHODS: AMI analogues resulting from (1) dealkylation, (2) deiodination and (3) deamination were synthesised and used as inhibitors in an in vitro biotransformation reaction of thyroxine (T4) to 3,3',5'-triiodothyronine (T3). Using high-performance liquid chromatography and ultraviolet detection for quantifying T3, it was found that the 5'-T4 deiodinase type I was involved in the reaction. On separate occasions, AMI or an AMI analogue was added to the reaction as an inhibitor. RESULTS: All studied AMI analogues inhibited 5'-T4 deiodination competitively (Ki value range 25-360 microM). In the concentration range of 1 1000 microM, AMI and its N-desethylated, deiodinated analogues inhibited 5'-T4 deiodination very weakly. AMI analogues with a hydroxyl group at the 4-position were strong inhibitors. Moreover, diiodo-AMI analogues inhibited 5'-T4 deiodination more strongly than their corresponding monoiodo- or deiodinated derivatives. CONCLUSION: It is likely that the degraded products of AMI could be responsible for thyroid dysfunction toxicosis in AMI therapy. PMID- 10805059 TI - Selegiline treatment and the extent of degenerative changes in brain tissue of patients with Alzheimer's disease. AB - BACKGROUND: A beneficial effect of selegiline (L-deprenyl) in Alzheimer's disease (AD) has been reported in several clinical studies. METHODS: The brain tissue from 17 deceased patients, members of a double-blind clinical trial to assess the potential benefit of selegiline in AD, were analysed. FINDINGS: In our study, the decrease in the Mini-Mental State Examination (MMSE) scores during the progress of the disease had been significantly influenced by selegiline treatment. Prior to death, the MMSE scores were significantly higher in those patients receiving selegiline than in those receiving placebo. However, according to our results, none of the lesions critical for AD diagnosis, such as counts of senile/neuritic plaques, neurofibrillary tangles or beta-A4 load, were influenced by the selegiline treatment. INTERPRETATION: In conclusion, according to our study, mechanisms other than neuronal degeneration seen as lesions critical for AD diagnosis are influenced by selegiline treatment, leading to the functional benefit found in AD. PMID- 10805060 TI - CYP2C19 polymorphism effect on phenobarbitone. Pharmacokinetics in Japanese patients with epilepsy: analysis by population pharmacokinetics. AB - OBJECTIVE: The aim of this study was to clarify the effect of genetic polymorphisms of CYP2C19 on the pharmacokinetics of phenobarbitone (PB) using a nonlinear mixed-effects model (NONMEM) analysis in Japanese adults with epilepsy. METHODS: A total of 144 serum PB concentrations were obtained from 74 subjects treated with both PB and phenytoin but without valproic acid. All patients were classified into three groups by CYP2C19 genotyping: G1, G2 and G3 were homozygous for the wild type of CYP2C19 (*1/*1), heterozygous extensive metabolizers (EMs), (*1/*2 or *1/*3), and poor metabolizers (PMs), (*2/*2, *2/*3), respectively. All data were analyzed using NONMEM to estimate pharmacokinetic parameters of PB with respect to the CYP2C19 genotype. RESULTS: Thirty-three patients belonged to G1 (44.6%), 35 to G2 (47.3%), and 6 to G3 (8.1%). The total clearance (CL) of PB significantly decreased by 18.8% in PMs (G3) relative to EMs (G1 and G2). The CL tended to be lower in G2 than in G1. CONCLUSION: In this study, we first demonstrated the effect of the CYP2C19 polymorphism on pharmacokinetics of PB by genotyping. The contribution of other metabolic enzymes in the metabolism of PB in humans remains to be elucidated; however, it appears that the disposition of PB is mediated in part by this enzyme. The estimated population clearance values in the three genotype groups can be used to predict the PB dose required to achieve an appropriate serum concentration in an individual patient. PMID- 10805061 TI - Simultaneous modelling of flosequinan and its metabolite. AB - DESIGN: A randomised, double-blind, prospective, placebo-controlled four-way study of the pharmacokinetics of single oral doses of flosequinan. We do not report the placebo data in this paper. Flosequinan was given at doses of 50, 100 and 150 mg, with a 2-week wash-out between periods. Blood samples were taken at a series of times up to 96 h after dosing. SETTING: Clinical pharmacology unit in a pharmaceutical company. PARTICIPANTS: Eighteen healthy volunteers of both genders, aged from 18 years to 55 years. MAIN OUTCOME MEASURES: Plasma concentrations of flosequinan and of its metabolite, flosequinoxan. RESULTS: We demonstrate that it is possible to model parent and metabolite concentration time profiles simultaneously and, in doing so, to estimate the first-pass effect using data from an oral administration. In our modelling approach, we propose a reasonably wide class of statistical models, allowing for left censoring. CONCLUSIONS: A parent-metabolite model that ignores the first-pass results in misleading predictions in a case where significant first-pass metabolism occurs. Thus, in phase-I studies, the new approach described in this paper can provide additional knowledge that may be useful in future formal studies. PMID- 10805062 TI - Pharmacokinetics and pharmacodynamics of the novel H1-receptor antagonist emedastine in healthy volunteers. AB - OBJECTIVE: Emedastine is a novel H1-receptor antagonist with pre-clinically well documented anti-allergic effects. Here, we set out to study the relationship between emedastine pharmacokinetics and the suppressive effect on histamine induced wheals and flares, and to compare these effects to placebo and cetirizine. METHODS: Emedastine (4 mg q.d.), emedastine (2 mg b.i.d.), cetirizine (10 mg q.d.) and placebo were administered to healthy volunteers in a double blind, cross-over fashion. On day 1 and day 5 (steady state) following drug administration, wheals and flares were induced by skin-prick testing with 1 mg ml(-1) or 10 mg ml(-1) histamine. RESULTS: Following the administration of 4 mg emedastine q.d., mean area under the concentration-time curve (AUC)0-24 values of 34.49 +/- 24.07 ng h ml(-1) and 47.05 +/- 36.12 ng h ml(-1) were attained on day 1 and day 5, respectively. Following the administration of emedastine (2 mg b.i.d.) mean AUC0-24 values were 29.75 +/- 19.92 ng h ml(-1) and 46.13 +/- 38.50 ng h ml(-1) on day 1 and day 5, respectively. Histamine-induced wheals and flares were significantly more effectively suppressed by emedastine and cetirizine than placebo. At pharmacokinetic steady-state levels, no significant difference could be found in the potency between cetirizine and emedastine (2 mg b.i.d.). CONCLUSION: Emedastine displays pharmacodynamic properties comparable with cetirizine and therefore qualifies as a safe and alternative compound with H1 receptor antagonist properties. Additional larger studies may be needed to substantiate potential benefits of cetirizine over emedastine after single-dose administration. PMID- 10805063 TI - Inhibition of human cytochrome P450 enzymes by 1,4-dihydropyridine calcium antagonists: prediction of in vivo drug-drug interactions. AB - OBJECTIVE: 1,4-Dihydropyridine calcium antagonists such as nifedipine are potent vasodilators. It is now commonly agreed that the oxidation of 1,4-dihydropyridine into pyridine, which is one of the main metabolic pathways, is catalysed by the cytochrome P450 (CYP) 3A4 isoform. In the present study, the inhibitory effects of 13 kinds of 1,4-dihydropyridine calcium antagonists clinically used in Japan on human CYP-isoform-dependent reactions were investigated to predict the drug interactions using microsomes from human B-lymphoblast cells expressing CYP. RESULTS: The specific activities for human CYP isoforms included 7-ethoxyresorfin O-deethylation (CYP1A1), phenacetin O-deethylation (CYP1A2), coumarin 7 hydroxylation (CYP2A6), 7-benzyloxyresorufin O-dealkylation (CYP2B6), S-warfarin 7-hydroxylation (CYP2C9), S-mephenytoin 4'-hydroxylaion (CYP2C19), bufuralol 1' hydroxylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and testosterone 6beta-hydroxylation (CYP3A4). Benidipine and amlodipine competitively inhibited the CYP1A1 activity. Nifedipine, nisoldipine and aranidipine competitively inhibited the CYP1A2 activity. No 1,4-dihydropyridie calcium antagonists used in this study inhibited the CYP2A6 activity. Barnidipine and amlodipine inhibited the CYP2B6 activity. Nicardipine, benidipine, manidipine and barnidipine competitively inhibited the CYP2C9 and CYP2D6 activities. Inhibition extent of the CYP2E1 activity by nifedipine and aranidipine were weak. Nicardipine, benidipine and barnidipine inhibited the CYP2C19 and CYP3A4 activities. Among the human CYP isoforms investigated, the inhibitory effects of 1,4-dihydropyridine calcium antagonists were potent on human CYP1A2, CYP2B6, CYP2C9, CYP2C19 and CYP2D6 as well as CYP3A4. Furthermore, the isoform selectivity of inhibition by 1,4-dihydropyridine calcium antagonists was clarified. CONCLUSIONS: In consideration of the Ki values obtained in the in vitro inhibition study and the concentration of 1,4-dihydropyridine calcium antagonists in human liver, the possibility of in vivo drug interactions of nicardipine and other drugs which are mainly metabolised by CYP2C9 and/or CYP3A4 was suggested. The inhibition of human CYP isoforms by 1,4-dihydropyridine calcium antagonists except nicardipine might be clinically insignificant. PMID- 10805065 TI - Circadian variation of the urinary 6beta-hydroxycortisol to cortisol ratio that would reflect hepatic CYP3A activity. AB - OBJECTIVES: Chronopharmacokinetics of drugs metabolized by cytochrome P450 3A (CYP3A) has been reported recently; however, little is studied on intra individual circadian variation in CYP3A activity in human. The aim of this study was to assess the intra-individual diurnal variation and day-to-day variation of the urinary 6beta-hydroxycortisol to cortisol ratio, a noninvasive index of human CYP3A activity. METHODS: Urine samples from ten healthy Japanese men were collected over four time intervals (0900 hours to 1300 hours, 1300 hours to 1700 hours, 1700 hours to 2100 hours and 2100 hours to 0900 hours) on days 1, 5 and 14 to verify diurnal variation, and 24-h urine was collected to study day-to-day variation over 2 weeks. Urinary 6beta-hydroxycortisol and cortisol were determined by means of high-performance liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry. RESULTS: The ratio of urinary 6beta-hydroxycortisol to cortisol exhibited noteworthy diurnal variation intra individually; 2.8-fold on average. However, day-do-day intra-individual variation of the ratio was not observed over 2 weeks; the coefficient of variation was 11.9 +/- 3.0%. CONCLUSION: The result indicates that imprudent use of random urine has a great risk of false evaluation in assessment of the 6beta-hydroxycortisol to cortisol ratio and that the ratio in 24-h urine samples provides a more robust measure of the inter-individual difference of this metabolic ratio, which to a certain but not complete extent represents the CYP3A activity. PMID- 10805064 TI - Human halothane metabolism, lipid peroxidation, and cytochromes P(450)2A6 and P(450)3A4. AB - OBJECTIVE: Halothane undergoes both oxidative and reductive metabolism by cytochrome P450 (CYP), respectively causing rare immune-mediated hepatic necrosis and common, mild subclinical hepatic toxicity. Halothane also causes lipid peroxidation in rodents in vitro and in vivo, but in vivo effects in humans are unknown. In vitro investigations have identified a role for human CYPs 2E1 and 2A6 in oxidation and CYPs 2A6 and 3A4 in reduction. The mechanism-based CYP2E1 inhibitor disulfiram diminished human halothane oxidation in vivo. This investigation tested the hypotheses that halothane causes lipid peroxidation in humans in vivo, and that CYP2A6 or CYP3A4 inhibition can diminish halothane metabolism. METHODS: Patients (n = 9 each group) received single doses of the mechanism-based inhibitors troleandomycin (CYP3A4), methoxsalen (CYP2A6) or nothing (controls) before a standard halothane anaesthetic. Reductive halothane metabolites chlorotrifluoroethane and chlorodifluoroethylene in exhaled breath, fluoride in urine, and oxidative metabolites trifluoroacetic acid and bromide in urine were measured for 48 h postoperatively. Lipid peroxidation was assessed by plasma F2-isoprostane concentrations. RESULTS: The halothane dose was similar in all groups. Methoxsalen decreased 0- to 8-h trifluoroacetic acid (23 +/- 20 micromol vs 116 +/- 78 micromol) and bromide (17 +/- 11 micromol vs 53 +/- 49 micromol) excretion (P < 0.05), but not thereafter. Plasma F2-isoprostanes in controls were increased from 8.5 +/- 4.5 pg/ml to 12.5 +/- 5.0 pg/ml postoperatively (P < 0.05). Neither methoxsalen nor troleandomycin diminished reductive halothane metabolite or F2-isoprostane concentrations. CONCLUSIONS: These results provide the first evidence for halothane-dependent lipid peroxidation in humans. Methoxsalen effects on halothane oxidation confirm in vitro results and suggest limited CYP2A6 participation in vivo. CYP2A6-mediated, like CYP2E1-mediated human halothane oxidation, can be inhibited in vivo by mechanism-based CYP inhibitors. In contrast, clinical halothane reduction and lipid peroxidation were not amenable to suppression by CYP inhibitors. PMID- 10805066 TI - Pharmacokinetics of a glycine site antagonist (gavestinel) following multiple dosing in patients with acute stroke. AB - OBJECTIVE: The objective of this study was to characterize the pharmacokinetics of gavestinel in patients with acute stroke. METHODS: Gavestinel was administered as an 800-mg loading dose and followed by either 100-, 200-, or 400-mg maintenance doses given every 12 h for five doses. Blood and urine samples were collected for pharmacokinetic evaluation. The pharmacokinetics of gavestinel were determined using compartmental analysis. RESULTS: The mean clearance (CL) and central (Vc) and steady-state (Vss) volumes of distribution across the dose groups were 0.31-0.40 l x h(-1), 3.3-3.9 l, and 9.8-17 l, respectively. The mean terminal half-life ranged from 29 h to 56 h. Gavestinel was extensively bound to plasma protein (median percentage free <0.01). During gavestinel administration, some patients exhibited elevated levels of bilirubin, which may be the result of shared mechanisms of elimination (glucuronide conjugation and excretion in bile). CONCLUSIONS: This study characterized the pharmacokinetics of gavestinel following multiple doses in acute stroke patients and showed that the pharmacokinetics are similar for increasing maintenance doses. The high protein binding of gavestinel was confirmed in acute stroke patients. A pharmacokinetic interaction between gavestinel and bilirubin may contribute to the increase in bilirubin. PMID- 10805067 TI - The impact of problem-based pharmacotherapy training on the competence of rational prescribing of Yemen undergraduate students. AB - OBJECTIVE: This study aims to reveal whether a short training course of problem based pharmacotherapy teaching, based on the World Health Organization's (WHO's) Guide to Good Prescribing and the Yemen Essential Drug List and Standard Treatment Guidelines, will improve the competence of rational prescribing among medical and health assistant students in Yemen. DESIGN: In a controlled pre/post test study, 111 students from universities and health institutes participated on a voluntary basis. They were randomly separated into a study and a control group. Students of the study group were taught to generate standard first-choice drugs for asthma or diarrhoea. The students were then taught how to apply this set of first-choice drugs to specific patient problems, using the WHO six-step problem solving approach. RESULTS: Students from the study group performed significantly better than those from control in all problems presented and also when compared with the results of the pre-test. The results of the pre-test also show that teaching students all basic knowledge about drugs does not guarantee rational prescribing in Yemen. CONCLUSION: It can be concluded from this study that proper training, i.e. 'immunising' future doctors using problem-based pharmacotherapy teaching, is an efficient way of teaching rational prescribing in Yemen. PMID- 10805068 TI - Influence of CYP2D6*2 and CYP2D6*4 alleles on phenotype in polymedicated depressed inpatients: therapeutic consequences? PMID- 10805069 TI - How did such a bad report get published in the EJCP? Trial of paracetamol, ibuprofen and nimesulide in children with upper respiratory infections (B. Ulukol et al., EJCP November 1999, 55: 615-618) PMID- 10805070 TI - Trk neurotrophin receptor-like proteins in the teleost Dicentrarchus labrax. AB - In recent years, data have accumulated suggesting that the role of neurotrophins and Trk receptors may not be limited to the nervous system, and the presence of these substances has been detected in a variety of vertebrate and invertebrate non-nervous tissues. This study was designed to map the expression of immunoreactivity (IR) for Trk-like proteins in alevins of the teleost Dicentrarchus labrax, with particular emphasis on non-nervous structures. We used antibodies against specific epitopes of the intracellular domain of these proteins, a region that is highly conserved in phylogeny. Trk-like IR was seen in segregate cell populations of the nervous system, and non-nervous tissues. In the central nervous system TrkA-like and TrkC-like IR was abundant, whereas TrkB-like IR was restricted to a low number of brain areas. Expression of Trk-like protein IR was observed in the peripheral nervous system and sensory organs, with the exception of the lateral line organ. Outside the nervous system, TrkA-like IR was mainly found in different epithelia, TrkB-like IR in the endocrine and digestive system, and TrkC-like IR in the cardiovascular and immune systems. The gills showed IR for all three Trk-like proteins, whereas they were absent from the gonads. Furthermore, scattered cells positive for Trk-like proteins were found in most of the investigated tissues. The distribution of Trk-like IR in this teleost is compared with that of mammals and birds, which it often paralleled, and the possible role of neurotrophins and Trk-like receptor proteins in different non neuronal tissues is discussed. PMID- 10805071 TI - Ependymal explants from the lateral ventricle of the adult bovine brain: a model system for morphological and functional studies of the ependyma. AB - By gently scraping off the surface of the lateral ventricles of adult bovine brains, we obtained sheets containing the ependymal layer and some attached sub ependymal cells. Explants were cultured in serum-free medium or in two media enriched with 20% fetal calf serum or 20% adult bovine cerebrospinal fluid, and processed for different time intervals from 4 h to 60 days. For characterization of the ependymal cells we used antisera against S-100 protein, vimentin and glial fibrillary acidic protein (GFAP). For comparison, the ependyma of adult bovines and of fetuses from days 60 to 120 post coitum was studied in situ. The adult ependyma consisted of a ciliated, cuboid cell monolayer with short basal processes; it displayed S-100 immunoreactivity but only scarce deposits of vimentin and no GFAP. The fetal ependyma had the appearance of a pseudostratified epithelium with elongated nuclei and basal processes containing S-100 and vimentin from day 80 post coitum and GFAP from day 100 post coitum. In explants, no differences were seen between the three culture media; the ependyma became pseudostratified, developed basal processes and showed increasing amounts of S 100 and vimentin first, and subsequently also GFAP. These changes were concomitant with the onset of mitotic activity in the subependymal layer leading to the production of numerous cells. The morphological and immunocytochemical features of ependymal cells in cultured explants resembled those of fetal ependyma. Our results indicate that the culture of ependymal explants from adult bovine lateral ventricles is an useful model system for morphological and functional studies of the ependyma and for the analysis of cell proliferation in the subependymal layer. PMID- 10805072 TI - First appearance and distribution of calretinin-immunoreactive neurons in the early development of the chick central nervous system. AB - Silver impregnation studies in chick embryos have shown that, by the 2nd day of incubation, the earliest neurofibrillar differentiation occurred in neuroblasts located at the diencephalic-mesencephalic junction and in the rhombencephalon; some of these neuroblasts were believed to become reticular neurons. Since calretinin, a cytosolic calcium-binding protein of the "E-F hand" family, occurs in reticular neurons, the present study investigated immunohistochemically whether the early differentiating reticular neurons are also the first neurons to express this marker during chick embryo development. The first calretinin immunoreactive neuroblasts appeared at stage 11 (40-45 h of incubation according to the series of Hamburger and Hamilton), and were located in the basal plate of the diencephalic-mesencephalic junction and of rhombomeres adjacent to the otic placode and in the alar plate and intermediate zone of the cervical spinal cord. In bromodeoxyuridine-injected embryos, these earliest calretinin-immunoreactive neurons were shown to express the calcium-binding protein 11-16 h after their last mitosis. By stage 11 up to the 14th day of incubation (stage 40), the calretinin-immunostained neurons increased in number and ultimately formed a chemically defined subset of neurons belonging to the tegmental reticular formation and raphe region of the brainstem. In the meantime, early calretinin immunostained nerve processes were shown to form two conspicuous longitudinal bundles which run in the ventral and lateral margins of the brainstem and spinal cord. PMID- 10805073 TI - Age-related changes in microtubules in the guinea pig organ of Corti. Tubulin isoform shifts with increasing age suggest changes in micromechanical properties of the sensory epithelium. AB - Biochemical and immunocytochemical analyses have been used to provide new insights into age-related changes in the sensory and supporting cells of the guinea pig organ of Corti. Quantitative densitometry of immunoblots showed that, while levels of alpha-tubulin remained relatively constant in guinea pigs from 3 weeks to 18 months old, there were progressive shifts in some tubulin isoforms. Levels of tyrosinated tubulin increased with age, nontyrosinatable tubulin (delta2-tubulin) showed a compensatory decrease, but detyrosinated tubulin did not change; acetylated, polyglutamylated, and glycylated tubulin levels also decreased. Immunolabeled tissue sections showed that cell type-specific distribution of tubulin seen in young guinea pigs (tyrosinated in the microtubules of the sensory cells, and post-translationally modified isoforms in the supporting cells) did not change as animals aged. However, there were age related decreases in labeling for alpha-tubulin and all post-translationally modified isoforms. Biochemical and immunocytochemical results both support an age related decrease in the number and/or length of microtubules as well as an increase in the pool of soluble tyrosinated and detyrosinated tubulin. They further suggest that microtubules containing nontyrosinatable tubulin from older animals are the sites for further modification of tubulin by acetylation, polyglutamylation, and glycylation. Changes in tubulin isoform levels and stability of microtubules in the organ of Corti may alter its micromechanical properties; the resulting changes in conduction of sound-induced vibration would provide one mechanism for age-related hearing loss. PMID- 10805074 TI - Intrinsic neurons in the rat ovary: an immunohistochemical study. AB - Previous studies have shown the presence of neuronal perikarya in the primate ovary, but not in the ovary from Sprague-Dawley rats. We report here that while such intrinsic neurons are indeed absent in this strain of rats, they can be visualized in the ovary from Wistar rats. The neurons, identified by their morphology and by the expression of NeuN (a neuron-specific nuclear protein), were detected at all postnatal intervals examined, from 14 h after birth to 50 days of age. While they were present in the ovarian hilum and medulla at all ages studied, neurons first appeared in the ovarian cortex during the juvenile period (postnatal days 10-20). In all cases, the size of the neuronal soma increased significantly during prepubertal development, reaching maximal values before puberty. Some neurons were catecholaminergic, as indicated by their content of immunoreactive tyrosine hydroxylase (TH), the rate-limiting enzyme of catecholamine biosynthesis. Some showed neuropeptide Y (NPY) immunoreactivity. TH positive neurons were seen either in isolation or clustered in ganglion-like structures in both the ovarian cortex and medulla. These results indicate that ovarian neurons are not present in all strains of rats, but when present, the chemical phenotype of some of them is of a sympathetic nature, similar to that described in primates. PMID- 10805075 TI - Differential spindle assembly checkpoint response in human lung adenocarcinoma cells. AB - CI-980 is an antimicrotubule agent that binds the colchicine site on tubulin. We examined CI-980 cytotoxicity in two lung adenocarcinoma cell lines, A549 and A427. Depolymerization of microtubules following CI-980 treatment resulted in a mitotic arrest in the A549 population, but not in the A427 population. Similar responses were obtained following treatment with Taxol and nocodazole. Drug treated A427 cells exited mitosis, generating a population dominated by multinucleated cells, while both multinucleated and apoptotic cells were present in the A549 population after extended drug treatment. CI-980-induced microtubule depolymerization was only partially reversible. However, regrowth of some microtubules in mitotic A549 cells following drug washout resulted in multinucleation of the population in the absence of apoptosis. These results show that A427 cells have a defective spindle assembly checkpoint. Levels of the MAD2 and BUB1 checkpoint proteins were similar in both A549 and A427 cells, suggesting that the checkpoint defect in the A427 cells is downstream of these proteins. In addition, induction of apoptosis in response to CI-980 correlates with the presence of a functional mitotic checkpoint and the extent of microtubule depolymerization. PMID- 10805076 TI - Differential effects of several retinoid receptor-selective ligands on squamous differentiation and apoptosis in airway epithelial cells. AB - The roles of the different retinoid receptors on the differentiation of rabbit tracheal epithelial (RbTE) cells in primary culture were analysed using selective agonists for the retinoid acid receptor subtypes RARalpha (CD336), RARbeta (CD2019), RARgamma (CD437), an RAR panagonist (CD367), a retinoid X receptor RXR panagonist (CD2624) and an antagonist for RARbeta/gamma (CD2665). Squamous differentiation was assessed via expression of cytokeratins CK13/CK4 and transglutaminase I (TGI), specific markers of metaplasia. Treatment with RARalpha and beta agonists or RAR panagonist, but not the RARgamma agonist or RXR agonist, is required for the inhibition of squamous metaplasia, evidenced by inhibition of CK13/CK4 and TGI expression. The expression of CK10 cytokeratin of keratinizing epithelia, CK14/CK5 basal cell cytokeratins, and CK6 marker of cell proliferation decreases upon exposure of the RARaalpha/beta and RXR agonists. The RARgamma agonist CD437, inactive in the decrease in CK13/CK4, CK10 and CK14, reduces CK5/CK6 amounts. CD437 is responsible for a dose-dependent apoptotic response. Nuclear labelling with propidium iodide (PI) and electron microscopy revealed chromatin condensation and nuclear fragmentation. DNA cleavage and cell fragmentation were confirmed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The RARbetagamma antagonist was also slightly active. The results indicate that CD437 causes growth arrest in the early S-phase of the cell cycle and prevents the transition G1-S-phase. CD437 was demonstrated to induce apoptosis in the S-phase cells identified by bromodeoxyuridine (BrdU) incorporation. In conclusion, RARalpha/beta ligands are effective inhibitors of squamous differentiation. On the contrary, RARgamma ligand appears to be inefficient in metaplasia inhibition, but the selective RARgamma agonist CD437 induces growth arrest and apoptosis of basal proliferative cells. PMID- 10805077 TI - Immunolocalization of vascular endothelial growth factor in the GH3 cell line. AB - The question of whether vascular endothelial growth factor (VEGF) is expressed in the GH3 cell line was investigated using immunocytochemistry, immunoelectron microscopy, and Western blotting. Using immunocytochemistry, VEGF was demonstrated in approximately 90% of the cells. Immunopositivity was localized mainly in the paranuclear Golgi region. In a small minority of cells, diffuse cytoplasmic immunostaining was also noted. By immunoelectron microscopy VEGF was evident in the secretory granules, cytoplasmic vesicles, rough endoplasmic reticulum, and the Golgi apparatus. Western blotting confirmed the results of the morphologic studies. It can be concluded that VEGF, which is know to induce angiogenesis and to increase vascular permeability, is produced in the prolactin- and growth hormone (GH)-secreting GH3 cell line. The functional role of VEGF in the GH3 cells is unknown. It is possible that this growth factor affects endocrine activity of GH3 cells by a paracrine mechanism. PMID- 10805078 TI - Differential expression of TGF-beta isoforms during postlactational mammary gland involution. AB - After cessation of lactation, the mammary gland undergoes involution, which is characterized by a massive epithelial cell death and proteolytic degradation of the extracellular matrix. Whereas the expression patterns and also the function of TGF-beta isoforms during mammary gland branching morphogenesis and lactation are well understood, their expression during postlactational involution and therefore a possible role in this process is poorly known. In this study we show that TGF-beta3 expression is dramatically induced (>fivefold) during mouse mammary gland involution when compared to that of virgin mouse, reaching a maximal expression level at day 4 after weaning. In contrast, other TGF-beta isoforms do not display significant increase in expression during involution (TGF beta1, 1.3-fold and TGF-beta2, <1.5-fold) when compared to that of virgin or lactating mice. During mammary gland involution, TGF-beta3 is expressed in the epithelial layer and particularly in myoepithelial cells. A comparison of the kinetics of TGF-beta3 expression to that of programmed cell death and degradation of the basement membrane suggests that TGF-beta3 functions in the remodeling events of the extracellular matrix during the second stage of involution. PMID- 10805079 TI - Coordinate expression of NGF and alpha-smooth muscle actin mRNA and protein in cutaneous wound tissue of developing and adult rats. AB - Nerve growth factor (NGF) is synthesized in cutaneous wound tissue, and its higher levels in the neonate may contribute to more efficient wound healing. We used in situ hybridization and immunohistochemistry to define NGF mRNA and protein expression in intact skin and following excision wounding in neonatal and adult rats. To determine whether NGF is associated with wound contractile fibroblasts (myofibroblasts), we also examined expression of alpha-smooth muscle actin (alpha-SMA) mRNA and protein, established markers for these cells. In intact skin, NGF mRNA and protein were present in vascular and arrector pili smooth muscle, hair follicle sheath cells, keratinocytes, and hypodermal fibroblasts. Neonatal adipocytes and Schwann cells also expressed NGF mRNA and protein, while adult adipocytes and Schwann cells displayed only NGF-ir. Following wounding, NGF mRNA expression was exuberant in these cell types, and increased similarly at both ages and appeared de novo in skeletal muscle cells. Additionally, both NGF mRNA and protein were present in macrophages and myofibroblasts, and expression in myofibroblasts was significantly greater in neonates. Wound myofibroblasts also expressed alpha-SMA. Surprisingly, after wounding alpha-SMA mRNA and protein were present in essentially all cells in which NGF mRNA was detected. We conclude that NGF expression is enhanced in many cell types after wounding, but greater NGF synthesis in neonates appears to be due to a more robust myofibroblast response. In addition, cell types which demonstrated NGF mRNA also expressed alpha-SMA, and staining for both markers increased following wounding, suggesting synthesis of both proteins is regulated in a coordinated fashion. PMID- 10805080 TI - Acute-phase response and circadian expression of connexin26 are not altered in connexin32-deficient mouse liver. AB - In mouse hepatocytes, the gap junctional proteins connexin32 (Cx32) and connexin26 (Cx26) are expressed in the same gap junctional plaque. Expression of the major Cx32 protein is downregulated during liver regeneration and cholestasis. Here we have analyzed the acute-phase response (after experimental inflammation) and circadian connexin expression in Cx32-deficient and wild-type mouse liver. Acute-phase response was triggered by intraperitoneal injection of lipopolysaccharide (LPS). Injection of recombinant mouse interleukin-1beta (mIL 1beta), mIL-6 or tumor necrosis factor alpha (mTNF-alpha) had no inflammatory effect. Northern blot analysis of positive and negative acute-phase transcripts following stimulation with cytokine or LPS revealed no difference between Cx32 deficient livers and wild-type controls, suggesting that loss of the Cx32 gene had no effect on experimental liver inflammation. Actin, beta-fibrinogen and Cx26 transcripts were increased after endotoxin stimulation. Under conditions of hepatic acute-phase response, Cx32 transcripts were not detected in LPS-treated livers of wild-type mice. Immunoblot analysis of proteins from inflamed wild-type livers indicated a strongly diminished amount of Cx32 protein, whereas the level of Cx26 protein was increased. Although intraperitoneal injection of mIL-1, mIL-6 as well as mTNF-alpha did not induce an acute-phase response, Cx32 protein expression was diminished, suggesting that post-transcriptional downregulation of Cx32 preceded the acute-phase response. Northern blot hybridization of RNA from wild-type and Cx32-deficient mouse liver revealed a similar circadian regulation of Cx26 and GAPDH transcripts with maximal expression around 2 p.m. and a minimum after midnight. PMID- 10805081 TI - Differentiation of rat thymic myoid progenitor cell line established by coculture with human T-lymphotropic virus type-I producing human T cells. AB - A thymus-derived myoid precursor cell line (ST1), which differentiates to myoid cells in the growth arrest condition, was established by the cocultivation of F344 rat thymic cells with human T-lymphotropic virus type-I (HTLV-I)-producing human lymphoid cells. No integration of HTLV-I was detected in ST1 cells by Southern blot hybridization. In a differentiation culture condition such as confluent culture or serum starvation, ST1 cells began to fuse, creating multinuclear giant cells, with the induced expression of MyoD1 and various muscle specific antigens, including alpha-sarcomeric actin, skeletal muscle myosin, myoglobin, desmin, and acetylcholine receptor. Ultrastructural investigation revealed that differentiated ST1B cells created aggregates of thick and thin filaments with Z-band-like composition, then formed sarcomeric structures and tubular honeycomb arrays. Finally, these cells spontaneously contracted with a frequency of 0.5-2.0 Hz and synchronized with adjoining cells. Transplantation of ST1B cells into nude mice produced a small tumor nodule, showing clear differentiation to skeletal muscle cells. ST1B cells did not indicate any colony forming activities in soft agar, demonstrating that ST1B cells retain some of the physiologically normal phenotypes. This rare cell line is promising for use in various physiological and pathological investigations including functional research of thymic myoid cells and the pathological role in autoimmune diseases, as well as animal model experiments of cell therapy related to muscular degenerative disorders or regeneration of injured muscles. PMID- 10805082 TI - Differential expression of mouse laminin gamma2 and gamma2* chain transcripts. AB - Laminins are large heterotrimeric basement membrane proteins that consist of alpha, beta, and gamma chains. We have previously shown that the human gamma2 and gamma2* transcripts result from the alternative use of the LAMC2 gene 3'-end exons. To explore the biological significance of the alternative gamma2 transcripts, we isolated the cDNA coding for the mouse laminin gamma2* transcript, characterized the 3'-end of the murine LAMC2 gene, and studied the expression of alternative gamma2 transcripts in several mouse tissues. The sequence reported here is the first one containing a full-length gamma2* 3'-UTR from any species. The mouse gamma2* transcript is 4110 bases and encodes a putative polypeptide of 1110 amino acids. This polypeptide lacks the C-terminal cysteine residue thought to be important for heterotrimer formation. The mouse gamma2* transcript was found to be expressed in several tissues by polymerase chain reaction (PCR), but at very low levels. The clearest signals were obtained on embryonic day 7, and in heart and testis of adult tissues. When the laminin gamma2* transcript expression pattern was compared with that of the gamma2 chain, a similar tissue distribution was observed. There was, however, a significant difference in expression levels. The longer gamma2 transcript was found to be much more abundant than the shorter gamma2* variant. Moreover, by whole-mount in situ hybridization, the shorter gamma2* form was localized in the mesenchyme of the developing kidney whereas the longer gamma2 form was exclusively present in the epithelium of the Wolffian (nephric) duct and ureteric bud. The results indicate different functions for the gamma2 variants. PMID- 10805083 TI - Distribution of gastrin-releasing peptide immunoreactivity in the brain of the collared dove (Streptopelia decaocto). AB - The distribution of immunoreactivity after applying an antibody against gastrin releasing peptide (GRP) was studied in the brain of the collared dove (Streptopelia decaocto). In the forebrain GRP-immunoreactive (GRP-ir) cells were found in the hyperstriatum accessorium, medial and lateral parts of the neostriatum, corticoidea dorsolateralis and temporoparieto-occipitalis areas, hippocampus, pre- and parahippocampal areas and prepiriform cortex. In the brainstem, GRP-ir cells were restricted mainly to the substantia nigra and ventral tegmental nucleus. Areas with densely packed GRP-ir clusters of varicosities were the medial intermediate hyperstriatum ventrale and lateral septal nucleus; dense GRP-ir neuropil was found in the parolfactory lobe, and in the dorsal half of the intermediate and caudal archistriatum. The ventral lamina medullaris contained many GRP-ir fibers. Forebrain areas devoid of immunoreactivity were the basal nucleus, ectostriatum, rostral archistriatum, most of the paleostriatum augmentatum and the lateral bed nucleus of the stria terminalis. Moderate densities of GRP-ir elements were found in the other telencephalic areas and further in, among others, the preoptic and hypothalamic region, ventral area of Tsai, cerulean nuclei, parabrachial complex, dorsal glossopharyngeal and vagus motor nuclei and medial nuclei of the solitary complex. The observations are compared with data from the literature and the implications for the definition of specific centers within the avian brain are discussed, with emphasis on systems with a role in visceral and motivational functions and in learning. PMID- 10805084 TI - Triple co-localisation of two types of allatostatin and an allatotropin in the frontal ganglion of the lepidopteran Lacanobia oleracea (Noctuidae): innervation and action on the foregut. AB - The triple co-localisation of peptidergic material immunoreactive to antisera raised against allatostatins of the Y/FXFGL-NH2 type, Manduca sexta allatostatin (Mas-AS), and allatotropin has been demonstrated in a single pair of anterodorsal neurones in the frontal ganglion of the tomato moth, Lacanobia oleracea (Noctuidae). Another pair of posterior neurones contain only Y/FXFGL-NH2-type allatostatin immunoreactivity. The neurites of all four cells trifurcate, and axons project to the brain in the frontal connectives and to the foregut in the recurrent nerve. Axons from the anterior neurones, within the recurrent nerve, have prominent lateral branches supplying muscles of the crop, and axons from both anterior and posterior cells show profuse branching and terminal arborisations in the region of the stomodeal valve. The brain contributes Y/FXFGL NH2-immunoreactive material, but not allatotropin or Mas-AS, to the recurrent nerve via NCC 1+2 and NCC 3. All three peptides have a reversible effect on the spontaneous (peristaltic) contractions of the foregut (crop) in vitro. Thus, both types of allatostatin are inhibitory at 10(-12) to 10(-7) M, whereas allatotropin is strongly myostimulatory at 10(-14) M. This is the first demonstration of the gut myoinhibitory effects of Mas-AS and, taken together with the effects of Y/FXFGL-NH2-type allatostatins and allatotropin, reveals a different functional aspect to that normally attributed to these three peptides, i.e. control of juvenile hormone synthesis by the corpus allatum. PMID- 10805085 TI - Mosaic arrangement of SCP(B-), FMRFamide-, and histamine-like immunoreactive sensory hair cells in the statocyst of the gastropod mollusc Pleurobranchaea japonica. AB - A pair of statocysts are located in the periganglionic connective tissue of the pedal ganglia of the opisthobranch mollusc Pleurobranchaea japonica. Light- and electron-microscopic observations show that the sensory epithelium of the statocyst consists of 13 disk-shaped hair cells. Each hair cell sends a single axon to the cerebral ganglion through the static nerve. Neurotransmitters in the hair cells were examined by means of immunocytochemistry. Our results show that the 13 sensory hair cells include two SCPB-, three FMRFamide-, and eight histamine-like immunoreactive cells. One hair cell contains a transmitter substance other than SCPB-, FMRFamide, histamine, serotonin, or GABA. One of the two SCPB-like immunoreactive cells, located in the ventral region of the statocyst, is the largest cell in the statocyst. The other, located in the anterodorsal region, shows co-immunoreactivity to both SCPB and FMRFamide antisera. Among the three FMRFamide-like immunoreactive hair cells, one is located in the posteroventral region, separated from the other two, which are adjacent to each other in the anterodorsal region. All the eight histamine-like immunoreactive hair cells are adjacent to one another, occupying the remainder of a triangular pyramid-shaped region. These immunoreactive cells are symmetrically placed in the right and left statocysts. This mosaic arrangement was identical among specimens. Thus the static nerve may code information about position or movement of the statoliths, with the use of different transmitters in the mosaic arrangement of the hair cells. PMID- 10805086 TI - Tracer studies of food absorption in the digestive tract of Nautilus pompilius (Cephalopoda, Tetrabranchiata). AB - In Nautilus pompilius, tracer experiments with 14C-labelled food show that the midgut gland, caecum and crop are involved in absorption of nutrients. According to liquid scintillation and light- and electron-microscopic autoradiography, the midgut gland exhibits the highest activity, followed by the caecum and crop. The density of silver precipitates is highest in the terminal alveoli of the midgut gland. Precipitates are also seen in the main cells of the caecal epithelium. Few precipitates are found in the lamina epithelialis mucosae of the crop, indicating that, in addition to food storage, digestive processes begin in this organ. These results have been confirmed by injection of the protein ferritin into the buccal cavity. The largest amount of ferritin is seen in the dense bodies of the main cells of the midgut gland, whereas those of the main cells of the caecum and crop contain less ferritin. PMID- 10805087 TI - Expression patterns of two putative odorant-binding proteins in the olfactory organs of Drosophila melanogaster have different implications for their functions. AB - The aqueous medium bathing the dendrites of olfactory neurons contains high concentrations of odorant-binding proteins (OBPs) whose role is still unclear. OBPs may facilitate interactions between odorants and their membrane-bound receptors, perhaps by increasing the water solubility of hydrophobic molecules. Alternatively, OBPs may be involved in the inactivation of odorants and other volatile molecules, preventing desensitization and/or protecting olfactory neurons from toxic chemicals. We report here novel features of the localization of two putative OBPs, PBPRP2 and PBPRP5, that have important and different implications for their role in olfaction. Unlike several other putative OBPs of Drosophila melanogaster that are only found in adult olfactory organs, PBPRP5 is also expressed in the larval olfactory organs, suggesting that it plays a common role in olfaction at both stages. In the adult, PBPRP5 expression is restricted to the sensillum lymph that bathes the olfactory dendrites of a subset of olfactory hairs, the basiconic sensilla. Since individual basiconic sensilla differ in olfactory specificity, PBPRP5 may be able to bind to and mediate olfactory responses to a wide range of odorants. In contrast, PBPRP2 is present in the space immediately below the antennal cuticle and in the outer cavity of approximately 30% of the double-walled coeloconic sensilla on the antennal surface. In neither case is PBPRP2 in contact with the dendritic membranes of olfactory neurons, making a carrier function unlikely for this protein. Instead, PBPRP2 may act as a sink, binding to odorants and other volatile chemicals and limiting their interactions with olfactory neurons. PMID- 10805088 TI - Endocrine cells in the gut of the shore crab Carcinus maenas immunoreactive to crustacean hyperglycaemic hormone and its precursor-related peptide. AB - The distribution and morphology of gut endocrine cells, which are immunoreactive to crustacean hyperglycaemic hormone (CHH) and the corresponding precursor related peptide (CPRP), have been described in the shore crab Carcinus maenas. The cells are uniquely distributed throughout the fore- and hindgut, but were never observed in the midgut or associated caeca. Expression of CHH and CPRP in the gut endocrine cells is generally restricted to premoult, although small numbers of immunoreactive cells were observed in intermoult and postmoult. A notable feature of the distribution of these slender cells was that, whilst they are distributed evenly over much of the fore- and hindgut, all extrinsic and intrinsic muscles of the gastric and pyloric stomach examined were surrounded by a ring(s) of cells, suggesting a mechanoreceptive function. Ultrastructural studies revealed that these cells contain numerous immunopositive, electron-dense granules. This suggests that they are "paraneurones", which secrete CHH and CPRP into the haemolymph during ecdysis, accounting for the ecdysial surge in CHH, which is implicated in water uptake and swelling prior to ecdysis. PMID- 10805089 TI - Axonopathy and amyotrophy in mice transgenic for human four-repeat tau protein. AB - Coding region and intronic mutations in the tau gene cause frontotemporal dementia and parkinsonism linked to chromosome 17. Some of these mutations lead to an overproduction of tau isoforms with four microtubule-binding repeats. Here we have expressed the longest four-repeat human brain tau isoform in transgenic mice under the control of the murine Thy1 promoter. Transgenic mice aged 3 weeks to 25 months overexpressed human tau protein in nerve cells of brain and spinal cord. Numerous abnormal, tau-immunoreactive nerve cell bodies and dendrites were seen. In addition, large numbers of pathologically enlarged axons containing neurofilament- and tau-immunoreactive spheroids were present, especially in spinal cord. Signs of Wallerian degeneration and neurogenic muscle atrophy were observed. When motor function was tested, transgenic mice showed signs of muscle weakness. Taken together, these findings demonstrate that overexpression of human four-repeat tau leads to a central and peripheral axonopathy that results in nerve cell dysfunction and amyotrophy. PMID- 10805090 TI - Atypical teratoid/rhabdoid tumor of the central nervous system: a comparative study with primitive neuroectodermal tumor/medulloblastoma. AB - Eleven atypical teratoid/rhabdoid tumors (AT/RT) and 121 primitive neuroectodermal tumors/medulloblastomas (PNET/MB) were included in this study for evaluation of the histopathological features of AT/RT and comparison between AT/RT and PNET/MB. Histopathological studies of AT/RT showed that in addition to the commonly recognized components, i.e., rhabdoid cells, small (PNET/MB) cells, spindle cells and epithelial components, there was a previously unrecognized component, sickle-shaped embracing cells, which were present in all cases and could be useful as a histological marker of this tumor. Immunohistochemical studies showed divergent differentiation of the tumor cells and among the 16 antibodies studied, vimentin, neuron-specific enolase, epithelial membrane antigen and glial fibrillary acidic protein were most commonly reactive. The frequency of AT/RT expressed as a ratio of AT/RT to PNET/MB was 1:11 in general and increased to 1:3.8 among patients younger than 3 years old. The AT/RT patients were younger than those with PNET/MB and had a female predominance. The MIB-1 labeling index of AT/RT was significantly higher than that of PNET/MB (mean 63.9 vs 40.1), which correlated with a shorter survival in patients with AT/RT than those with PNET/MB (median survival time 15.4 months vs 156.4 months). PMID- 10805091 TI - Parkinson's disease: affection of brain stem nuclei controlling premotor and motor neurons of the somatomotor system. AB - Pathological changes which consistently develop in the lower brain stem of patients suffering from Parkinson's disease are described against the background of the internal organization and interconnections of the involved nuclei, i.e., the gigantocellular reticular nucleus, bulbar raphe nuclei, and coeruleus subcoeruleus area. Immunoreactions against the presynaptic protein alpha synuclein reveal not only the voluminous forms of Lewy bodies and Lewy neurites but also the otherwise inconspicuous dot- or thread-like types. These lesions develop solely in specific neuronal types. Lipofuscin- or neuromelanin-laden projection cells which at the same time generate a long, unmyelinated or sparsely myelinated axon are particularly susceptible to developing the changes. The bulbar nuclei under consideration receive strong input from supramedullary sources, above all from higher order centers of the limbic system such as the central amygdalar nucleus, periaqueductal gray, and parabrachial nuclei. In turn, they generate descending projections to premotor and motor neurons of the somatomotor system. The disease-related deterioration of both the supramedullary limbic centers and the bulbar brain stem nuclei reduces the limbic influence and markedly impairs the control of premotor and motor neurons. This functional deficit most probably contributes to the overall dysfunction of the motor system typically evolving in the course of Parkinson's disease. PMID- 10805092 TI - Glycogenosomes in the aging rat brain: their occurrence in the visual pathways. AB - The significance of glycogenosomes (glycogen bodies), frequently seen in peripheral neurites of aging rats, is unknown and their occurrence elsewhere in nervous tissue is poorly documented. During the course of another study these bodies were observed by light microscopy in the visual pathways of aging rats where they have not previously been noted, and this report documents their occurrence, localisation and changes in density with age. Using the periodic acid Schiff stain, small brightly red-staining bodies, digested by diastase and containing beta-glycogen particles, were seen in increasing numbers in the neuropil of the superior colliculi in brain sections from animals of 5 months of age onwards. From 1 year until more than 2 years of age they steadily became more numerous in the outer one third of the superior colliculus, but remained small, rarely exceeding 4 microm. They were also found at later times in small numbers lying singly in the optic tract, the optic chiasm and optic nerves, although rarely in lateral geniculate nuclei. Similar bodies were also found to accumulate with age in the retinal photoreceptor cell layer. Changes in their densities and size with age in both regions have been documented and it is suggested that, while their occurrence in retinal photoreceptor cells may be due to sustained light damage leading to mitochondrial oxidative stress, it is difficult to implicate this mechanism for their occurrence in retino-tectal nerve fibres. The role of physical trauma, suggested for the presence of these bodies in aging peripheral axons, can be excluded and they appear not to be related to polyglucosan bodies. PMID- 10805093 TI - Simultaneous loss of hamartin and tuberin from the cerebrum, kidney and heart with tuberous sclerosis. AB - Tuberous sclerosis (TSC) is caused by a mutation in either the TSC1 or TSC2 gene. The clinical manifestations of mutations of the two genes are hardly distinguishable, for reasons as yet unknown. In this study, we examined the expression of the products of these genes, hamartin and tuberin, in control and TSC tissues. Western blotting disclosed that hamartin and tuberin are both abundant in the cerebral gray matter and that they have similar subcellular distributions and developmental patterns of expression. Immunohistochemical localizations of hamartin and tuberin were also similar, with high levels of expression being localized to the cerebral neurons and glial cells, renal uriniferous and collecting tubules, and cardiac muscles. In the cerebrum with TSC, both hamartin and tuberin were simultaneously reduced in the cortical tubers and subependymal giant cell astrocytomas, and from the normal-appearing cortex. The renal angiomyolipomas and cardiac rhabdomyomas also showed a loss of both the proteins. These results provide evidence for the co-localization and interaction of hamartin and tuberin in vivo, and suggest that a mutation in one TSC gene may secondarily affect the expression of the other in some TSC lesions. PMID- 10805094 TI - Inconsistency between severe substantia nigra degeneration with Lewy bodies and clinical parkinsonism in dementia patients: a cliniconeuropathological study. AB - In a retrospective cliniconeuropathological study, we reviewed all the cases received in our dementia brain bank during a 4-year period to determine if all patients with severe substantia nigra (SN) degeneration and SN Lewy bodies (LBs) exhibited prominent signs of parkinsonism and were treated for parkinsonism during the disease course. The SN of 426 cases were graded for microscopic degeneration using a semiquantitative five-tiered scale, with grade 0 indicating normal and grade 4 the most severe degeneration. Twenty-nine cases with grade 3 (16) or grade 4 (13) SN degeneration with SN LBs and clinical records were identified. Ten had been treated for parkinsonism (6 grade 3, 4 grade 4) and 19 had not. Whereas most of the patients had exhibited signs of end-stage parkinsonism during their last year, 1 grade 3 and 2 grade 4 patients apparently never exhibited prominent signs of parkinsonism during the course of their dementia. No clear neuropathological differences were noted between these patients that did not have prominent signs and a control group of six patients with clinical Parkinson's disease with dementia (parkinsonism onset at least 1 year before dementia onset). We conclude that in patients with dementia there is an inconsistent relationship between the expression of clinical parkinsonism during life and severe SN degeneration with LBs identified at necropsy. PMID- 10805095 TI - Altered calcium homeostasis in spinal motoneurons but not in oculomotor neurons of SOD-1 knockout mice. AB - SOD-1-deficient mice demonstrate no loss of motoneurons but are still vulnerable to axotomy and ischemic insults. To investigate possible reasons for vulnerability of motoneuron populations, we studied changes in ultrastructural calcium distribution during maturation in spinal- and oculomotor neurons in SOD 1(-/-) mice. Between 3 and 11 months the cytoplasmic component of the intracellular calcium changed at a lower rate in spinal motoneurons and motor axon terminals in the interosseus muscle of SOD-1(-/-) animals compared to wild type controls. No such dissimilarities were noted in the oculomotor system, or in mitochondrial calcium contents of either cell type. These data suggest that the lack of SOD-1 may be associated with vulnerability to insult by depletion of non mitochondrial calcium stores selectively in motoneurons lacking parvalbumin and/or calbindin D28K. PMID- 10805096 TI - Tubulovesicular particles occur early in the incubation period of murine scrapie. AB - Tubulovesicular bodies are structures, apparently specific to the transmissible spongiform encephalopathies, which are of unknown composition and significance. Prion protein (PrP) is absent from tubulovesicular bodies when tissues are examined by immunogold electron microscopy. In the F1 cross of C57 and VM mice (CVF1) infected with ME7 scrapie there is a marked degeneration of hippocampal CA1 neurons. In this model the earliest changes seen, at about 100 days post inoculation (dpi) are a degeneration of axon terminals and synaptic loss. Terminal disease is around 250 dpi. In blind coded trials we counted the number of tubulovesicular particles and estimated their density in 56-76 electron micrographs taken from the stratum radiatum of each of one or two CVF1 ME7 infected mice at 84, 100, 126, 154 and 181 dpi and from four normal brain inoculated control mice. Tubulovesicular particles were present from 98 dpi and the density of particles increased with increasing incubation period. The very early occurrence of tubulovesicular particles, before the presence of significant pathology, argues that tubulovesicular particles are a part of the primary disease and are not epiphenomena. PMID- 10805097 TI - No loss of hippocampal neurons in AIDS patients. AB - Acquired immunodeficiency syndrome (AIDS) is often accompanied by neurological disorders and neuropathological abnormalities. A loss of up to 30% of the neocortical neurons in brains from AIDS patients has been demonstrated in several studies. This neuron loss appeared to have no relationship with either the presence of clinical dementia or HIV encephalitis. The hippocampal region of the brain is an essential component of learning and memory processes and it is severely affected in a number of diseases, including temporal lobe epilepsy and Alzheimer's dementia. In view of the reports of a considerable loss of neocortical neurons, the aim of the present study was to investigate the degree to which neurons may be lost in hippocampus of AIDS patients. A stereological method, the optical fractionator, was used to estimate the total number of neurons in the five subregions of the hippocampus of nine AIDS patients and ten controls. There were no statistically significant differences in the group means of the number of neurons in any of the subdivisions. This result is discussed in the light of the large loss of neurons in the neocortex reported to take place in AIDS patients. PMID- 10805098 TI - Modeling of a periodic instability in paired helical filaments reveals an axial repeat. AB - Ultrastructural studies of paired helical filaments (PHF) have been facilitated by the ability to isolate enriched fractions of detergent-insoluble forms of PHF. These fractions are composed of a relatively homogeneous population of short (usually < 400 nm) highly fragmented PHF. A small proportion of isolated PHF have highly stereotyped angled profiles that represent deformations due to structural instability. These distorted PHF can be characterized quantitatively using a simple numerical procedure that reveals that the axial instabilities occur with predictable regularity over the length of the PHF. Using a structural model of PHF, it is shown that the periodicity of the axial instability can be correlated to an axially repeated subunit of uniform size. The upper limit for the axial extent of the repeated segment was calculated to be 80 nm, similar to the size of a single one-half twist in the PHF ribbon. It is proposed that this segment may represent one type of particle in the hierarchy of structural subunits in the PHF ribbon, or an oligomeric intermediate species in PHF assembly. PMID- 10805099 TI - The extracellular matrix of peripheral nerve in diabetic polyneuropathy. AB - The pattern of collagenisation in peripheral nerve in diabetic polyneuropathy was examined in nerve biopsy specimens from patients with diabetic polyneuropathy in comparison with organ donor control nerves and disease controls (other neuropathies). There was increased endoneurial collagenisation both in the diabetic polyneuropathy cases and the disease controls, this predominantly involving types I and III. Type II collagen was not detected in organ donor control nerves or in the diabetic and the disease control nerves. There was a relative increase in type VI collagen in the endoneurium in the diabetic nerves immediately surrounding groups of Schwann cells. This was not a feature in the other neuropathies. The quantity of types IV, V and VI collagen was increased around the endoneurial microvessels in the diabetic patients and, to a lesser extent, in those with hereditary motor and sensory neuropathy (HMSN). Increased deposition of types IV and V collagen was observed in the perineurium in the diabetic nerves, the latter being most evident in the innermost lamellae where the amount of laminin was possibly also increased. The diameter of the general endoneurial collagen fibrils was greater in the diabetic nerves, although this was not more than in a disease control (HMSN). The collagen fibrils that were present within the basal laminal tubes that had surrounded degenerated myelinated fibres in the diabetic nerves, and those within the onion bulbs of the HMSN cases, were of the normal endoneurial calibre. The expression of laminin by Bungner bands in diabetic neuropathy did not differ from that in disease control nerves, nor were any differences detected for fibronectin. Whether the changes observed are important for the impaired regenerative capacity in diabetic neuropathy requires further investigation. PMID- 10805100 TI - Temporal lobe epilepsy with and without psychosis: exploration of hippocampal pathology including that in subpopulations of neurons defined by their content of immunoreactive calcium-binding proteins. AB - We have investigated relationships between hippocampal/temporal lobe neuropathology and psychosis in subjects with temporal lobe epilepsy, paying particular attention to possible differences in density of hippocampal neurons immunoreactive for calcium-binding proteins. There was a trend for a greater prevalence of left handedness in the psychotic (n = 6) than the non-psychotic (n = 26) cases (P = 0.0504). Psychotic cases also differed from non-psychotic ones in having: (1) more focal lesions outside the hippocampus (P = 0.006); (2) less severe CAI neuron loss (P = 0.015); and (3) a trend, after Bonferroni correction, for a higher density of calbindin-immunoreactive neurons in the CA4 (P = 0.022). An additional finding was that dentate granule cell dispersion was significantly associated with the presence of a reduced density of calretinin-immunoreactive neurons in CA4 (P = 0.002) and with a more severe loss of CA4 neurons visible with Nissl stain (P = 0.003). Thus, cases of temporal lobe epilepsy with psychosis were distinguishable on the basis of a higher density of calbindin reactive neurons in CA4 as well as on more general aspects of their pathology. PMID- 10805101 TI - Evidence for a constitutive, verapamil-sensitive, non-P-glycoprotein multidrug resistance phenotype in malignant glioma that is unaltered by radiochemotherapy in vivo. AB - Human malignant gliomas are commonly resistant to chemotherapy. Here, we examined the role of the multidrug resistance (mdr) mechanism in the chemo-resistance of these tumors, using a twofold approach: (i) by assessing a possible mdr phenotype before and after chronic drug exposure of glioma cells in vitro, and (ii) by assessing the modulation of expression of the mdr-associated P-glycoprotein (Pgp) using radiotherapy and serial cycles of chemotherapy in human glioblastoma patients in vivo. T98G, and to a lesser degree, LN-229 human malignant glioma cells exhibit a constitutive mdr phenotype as determined by the modulation of dye transport and by the augmentation of chemosensitivity by the mdr antagonist, verapamil. Thus, coexposure to verapamil enhances the cytotoxicity of vincristine, doxorubicin and VM26 in T98G cells and that of vincristine in LN-229 cells. Chronic exposure of the cells to low concentrations of vincristine and doxorubicin, but not VM26, topotecan or BCNU, moderately enhances the mdr-like phenotype, as assessed by drug expulsion assays. However, chronic exposure to increasing drug concentrations does not significantly alter the sensitivity to the respective drugs. These data are consistent with a constitutive, but not drug inducible, mdr-like drug resistance in glioma cells in vitro. Immunocytochemical analysis of human malignant gliomas in vivo reveals that Pgp expression is more abundant in endothelial cells within the gliomas, than in the glioma cells proper. Importantly, Pgp expression is unaltered by radiochemotherapy, assessed by comparative immunocytochemistry of glioma specimens obtained serially before and after radiochemotherapy. We conclude that (i) glioma cells exhibit constitutive mdr-like drug resistance that is not significantly altered by chronic drug exposure in vitro; (ii) endothelial cells may play an important role in Pgp-mediated drug resistance of gliomas in vivo; (iii) radiotherapy and repeated chemotherapy cycles do not modulate Pgp expression in human malignant gliomas in vivo; (iv) there is preliminary evidence for a non-Pgp, verapamil sensitive drug transport activity in glioma cells. PMID- 10805102 TI - Gliomas of the optic nerve: histological, immunohistochemical (MIB-1 and p53), and MRI analysis. AB - Gliomas of the optic nerve, although typically of pilocytic (WHO grade I) histology, can present within the spectrum of astrocytic neoplasia including glioblastoma (WHO grade IV). In certain cases, histologic features alone make the distinction between pilocytic and diffuse astrocytomas difficult. We reviewed 22 cases of optic nerve gliomas, 19 of which were pilocytic astrocytomas (PA), and 3 of which were diffuse, non-pilocytic astrocytomas. The cases were evaluated for their clinical course, radiographic appearance, histologic grade, and proliferation indices as detected by MIB-1 (Ki-67) and p53 antibodies. Of the 19 PA, 14 showed no tumor growth by magnetic resonance imaging, and had Ki-67 and p53 labeling indices (LI) of < 1%. The other 5 PA exhibited aggressive behavior manifest by marked diffuse infiltrative tumor growth causing death in 2 patients, 1 of whom was diagnosed with neurofibromatosis type 1 (immunoperoxidase and radiographs not available), and marked local growth with an average time to growth of 39.3 months, a Ki-67 LI of 2-3%, and a p53 LI of < 1% in three others. Three of the five aggressive PA histologically demonstrated a finely reticulated pattern, a pattern that appears as an exaggeration or expansion of the normal neuroglia of the optic nerve, and may simulate a diffuse low-grade astrocytoma. Two demonstrated the coarsely reticulated pattern, with the biphasic and microcystic pattern typical of PA. Three diffuse astrocytomas (2 anaplastic astrocytomas and 1 glioblastoma) originated clinically and radiographically from the optic nerve, and revealed a Ki-67 LI of 2-12%, a p53 LI of 2-8%, and an average time to growth of 8 months. We conclude that the majority of PA of the optic nerve are non-aggressive, stabilize radiographically, and have Ki-67 and p53 LI < 1%. However, a subpopulation of PA has a propensity for aggressive behavior, and are identified by a Ki-67 LI of 2-3% and a p53 LI of < 1%. Diffuse astrocytomas have both Ki-67 and p53 LI > 2%. Thus, in cases of aggressive optic nerve tumors in which the histologic review of biopsy material cannot confidently confirm the diagnosis of pilocytic or diffuse fibrillary glioma, a p53 LI of > 1% appears to favor the diagnosis of diffuse astrocytoma. PMID- 10805103 TI - Unmyelinated nerve fiber degeneration in chronic inflammatory demyelinating polyneuropathy. AB - To determine whether unmyelinated nerve fibers escape degeneration as one might expect in an immune response exclusively directed at myelin, we performed a morphometric examination of unmyelinated axons and myelinated nerve fibers in sural nerve biopsy specimens of 14 patients with a chronic inflammatory demyelinating polyneuropathy (CIDP) and of 12 age-matched normal controls. The numbers of unmyelinated axons, myelinated nerve fibers, denervated Schwann cell units and collagen pockets were quantified and related to the clinical and electrophysiological data of the patients with CIDP. In 4 patients with a rapid onset of the neuropathy and a highly elevated CSF protein, the numbers of both unmyelinated axons and myelinated nerve fibers were decreased equally. In 8 patients we found that the unmyelinated axons were relatively spared compared with the loss of myelinated nerve fibers. In these patients, however, the presence of denervated Schwann cell units and of collagen pockets was increased. We conclude that unmyelinated nerve fibers are affected in patients with CIDP. PMID- 10805104 TI - Clinical validity of Braak neuropathological staging in the oldest-old. AB - Several studies have demonstrated a good correlation between clinical severity and Braak's neuropathological staging in Alzheimer's disease (AD). However, nonagenarians and centenarians display a different pattern of cortical vulnerability to the neurodegenerative process compared to younger elderly, and it is not known whether correlations between clinical severity and neuropathological stages remain valid in this age group. To address this issue we compared Clinical Dementia Rating scale (CDR) scores and Braak stages in 116 patients over 90 years of age with either no cognitive impairment or very mild to severe AD. There is a strong positive correlation between CDR scores and Braak staging (Spearman coefficient = 0.66; P < 0.01). However, neuropathological staging does not distinguish cases with normal cognition (CDR 0) from those with mild cognitive changes (CDR 0.5). Unlike younger cohorts, Braak stages I and II are frequently associated with questionable dementia in this age group. Braak stage III overlaps with all CDR levels and correlates poorly with cognitive function. Braak stages IV or greater are consistently associated with at least mild dementia. Consistent with our previous neuropathological analyses of nonagenarians and centenarians, the present data suggest that the substantial involvement of the hippocampus which characterizes Braak stage IV is a key step in the development of overt clinical signs of dementia in the oldest-old. Moreover, they indicate that Braak staging represents a broad concept of the evolution of neurofibrillary tangles rather than a precise hierarchical model associated with a stepwise deterioration of cognitive abilities near the upper limit of life. PMID- 10805105 TI - Tumor necrosis factor-alpha expression in muscles of polymyositis and dermatomyositis. AB - We evaluated the expression of tumor necrosis factor-alpha (TNF-alpha) mRNA in muscle biopsy specimens from patients with polymyositis (PM) and dermatomyositis (DM) to clarify its role in the pathogenesis of PM and DM. We performed non radioactive in situ hybridization studies for TNF-alpha combined with immunohistochemistry for cell type-specific markers on muscles from ten PM and five DM patients. TNF-alpha-positive infiltrating cells present in the endomysium and perimysium were found in all PM and DM muscles. The frequency of TNF-alpha positive cells against total infiltrating cells was similar among PM and DM (27.1 +/- 7.4% in PM and 28.5 +/- 13.6% in DM). However, TNF-alpha/CD8-positive lymphocytes and TNF-alpha-positive macrophages invading the non-necrotic muscle fiber were observed only in PM but not in DM. TNF-alpha was more highly expressed in PM and DM than was previously thought, and it was suggested that TNF-alpha plays a role in muscle fiber degeneration in PM. PMID- 10805106 TI - Neuronal intranuclear hyaline inclusion disease with polyglutamine-immunoreactive inclusions. AB - Neuronal intranuclear hyaline inclusion disease (NIHID) is a group of neurodegenerative disorders characterized by the presence of intranuclear inclusions in neurons (NIs). We report here clinicopathological findings of a 25 year-old female patient who died after 13 years of a clinical course characterized by progressive gait disturbance and movement disorders. Histological examination revealed widespread NIs with neuronal loss in restricted regions; neuronal loss was severe in the subthalamic nucleus, internal pallidum, substantia nigra, Edinger-Westphal nucleus and Purkinje cell layer. Quantification of the NIs combined with a graded evaluation of neuronal loss revealed an overall tendency for more severe neuronal loss to be accompanied by a lower frequency of NIs. A morphological similarity to the nuclear inclusions recently identified in several CAG repeat diseases prompted us to examine the immunolocalization of ubiquitin and expanded polyglutamine stretches, which demonstrated the presence of ubiquitin at the periphery of most NIs. An expanded polyglutamine stretch was seen in the center of limited number of NIs. These findings indicate that abnormal fragments such as expanded polyglutamine regions are incorporated into the inclusion, aggregated in its center, and thereby metabolized by a ubiquitin-dependent proteolytic pathway. Although it remains to be elucidated how the formation of NIs is related to neuronal degeneration, our findings suggest that NIs are formed in the process of sequestering or degrading abnormal protein fragments and formation of NIs may not be immediately toxic to neurons. PMID- 10805107 TI - Cerebral malakoplakia associated with Escherichia coli infection. AB - Malakoplakia is an unusual chronic inflammatory disease occurring predominantly in the bladder and only rarely affecting other organs. For the urinary tract, its aetiology has been ascribed to the presence of Escherichia coli, while the very few cases of cerebral malakoplakia which have been reported so far, have mostly occurred in infants in the clinical setting of neonatal herpes virus infection or otherwise in adults in areas of cerebral infarction. We here report a case of E. coli-associated malakoplakia of the brain. It occurred in a 53-year-old man who had undergone long-term corticosteroid therapy and had previously been operated on a cerebral E. coli-associated abscess. This case indicates that malakoplakia of the brain might also be a histiocytic reaction against bacterial antigens of the E. coli family. PMID- 10805108 TI - T cell-mediated paraneoplastic ganglionitis--an autopsy case. AB - A 57-year-old woman presented with subacute sensory, ataxic neuronopathy. Clinical investigation revealed a right-sided non-small-cell lung cancer. Serum investigation for specific antineuronal antibodies was negative. Histology showed T lymphocytic infiltrates in dorsal root ganglia. The observed histological pattern is similar to that described in antibody-positive cases. Thus, these findings suggest similar pathways in specific antineuronal antibody-negative and positive cases of paraneoplastic subacute sensory neuronopathy. PMID- 10805109 TI - Graves' ophthalmopathy: diagnosis and management. PMID- 10805110 TI - Radiation safety of the sentinel lymph node technique in breast cancer. AB - Many publications attest to the potential of the sentinel lymph node technique in advancing the clinical management of melanoma and, more recently, breast cancer. Whilst not yet universally regarded as the standard of care, the technique is gaining wide acceptance. Use of a radiolabelled colloidal tracer is central to optimising sensitivity, and this brings with it the need to address radiation safety issues relating to the use of radioactive materials in the operating theatre and pathology laboratory, and the generation of radioactive waste. The radiation dose to the patient should also be determined if the professional is to reassure the patient by placing this in its proper context. For the purpose of this investigation, biodistribution data were obtained from patient studies to quantify the migration of tracer beyond the injection site, thereby permitting a detailed assessment of the internal dosimetry of the tracer and the resulting radiation dose to the patient. Uptake of tracer in the sentinel nodes, reticulo endothelial system and circulating blood was investigated. The radiation dose to surgical staff was recorded using whole-body monitors and extremity dosimeters worn at the fingers. Clinical waste in the operating theatre was monitored and the radioactive content of significantly contaminated items determined. The radiation dose to pathology staff was estimated from knowledge of the radioactive content of the specimens obtained and a study of work practices. Migration of tracer was found to be minimal, with greater than 95% retention at the injection site. The effective dose resulting to the patient was 2.1x10(-2) mSv/MBq, with a mean breast dose of 7.2x10(-1) mGy/MBq. A mean whole-body dose of 0.34 microSv was received by surgical staff per procedure, with a mean finger dose of 0.09 mSv (90 microSv). Radiation doses received by pathology staff will be predominantly below measurable levels and are likely to be negligible unless primary specimens from a large number of studies are analysed promptly upon their excision. At operation, surgical swabs can become significantly contaminated and have been found to contain up to 22% of the administered activity, dependent upon the surgical procedure performed. It is concluded that moderate activities of technetium-99m labelled tracer are administered to the patient, and the radiation risk to the patient is consequently low relative to that from many other medical exposures. The radiation doses to staff groups involved in all aspects of the technique are low, and under normal circumstances and levels of workload, routine radiation monitoring will not be required. Standard biohazard precautions prevent direct intake of radioactive contamination. Radioactive waste is created in the operating theatre, and may be generated in the pathology laboratory if specimens are not routinely stored until fully decayed. This will require special handling if the disposal of radioactive material is not permitted. PMID- 10805111 TI - Scintigraphic detection of acute experimental endocarditis with the technetium 99m labelled glycoprotein IIb/IIIa receptor antagonist DMP444. AB - Bacterial endocarditis is an important clinical problem that may result in persistent bacteraemia and irreversible cardiac damage. Since endocarditis is characterized by aggregation of activated platelets, fibrin and bacteria, we studied DMP444, a technetium-99m labelled high-affinity antagonist of the GP IIb/IIIa receptor that is expressed on activated platelets. In seven Beagle dogs (11-15 kg), the left ventricle was catheterized via the right carotid artery. One hour later, 5x10(7) colony forming units of Staphylococcus aureus were injected intracardially. Half an hour later, the catheter was removed. Two extra dogs underwent a complete sham procedure. One day after the intervention, five infected and the two non-infected dogs were injected with 37 MBq/kg 99mTc-DMP444 and two infected dogs with 37 MBq/kg 99mTc-IgG (used as a non-specific control agent) and imaged up to 4 h after injection. Samples were obtained for tissue counting, microbiology and histology. From 1 to 2 h post injection onward, there was clear focal accumulation of DMP444 in the aortic valve region when endocarditis was present, and this accumulation increased with time. The non infected and the 99mTc-IgG injected dogs showed only persisting blood pool activity without any focal abnormality. At 4 h post injection, the in vivo valve to-blood pool ratios were 1.87+/-0.18 in endocarditis, 1.01+/-0.05 in non infected controls and 1.09+/-0.02 in 99mTc-IgG injected dogs (P<0.05). It is concluded that targeting activated platelets with the 99mTc-labelled GP IIb/IIIa antagonist DMP444 allows a final diagnosis of experimental bacterial endocarditis within 4 h owing to high, specific and fast in vivo uptake. PMID- 10805112 TI - Automated interpretation of ventilation-perfusion lung scintigrams for the diagnosis of pulmonary embolism using artificial neural networks. AB - The purpose of this study was to develop a completely automated method for the interpretation of ventilation-perfusion (V-P) lung scintigrams used in the diagnosis of pulmonary embolism. An artificial neural network was trained for the diagnosis of pulmonary embolism using 18 automatically obtained features from each set of V-P scintigrams. The techniques used to process the images included their alignment to templates, the construction of quotient images based on the ventilation and perfusion images, and the calculation of measures describing V-P mismatches in the quotient images. The templates represented lungs of normal size and shape without any pathological changes. Images that could not be properly aligned to the templates were detected and excluded automatically. After exclusion of those V-P scintigrams not properly aligned to the templates, 478 V-P scintigrams remained in a training group of consecutive patients with suspected pulmonary embolism, and a further 87 V-P scintigrams formed a separate test group comprising patients who had undergone pulmonary angiography. The performance of the neural network, measured as the area under the receiver operating characteristic curve, was 0.87 (95% confidence limits 0.82-0.92) in the training group and 0.79 (0.69-0.88) in the test group. It is concluded that a completely automated method can be used for the interpretation of V-P scintigrams. The performance of this method is similar to others previously presented, whereby features were extracted manually. PMID- 10805113 TI - Dynamic 2-[18F]fluoro-2-deoxy-D-glucose positron emission tomography of liver tumours without blood sampling. AB - Positron emission tomography (PET) using 2-[18F]fluoro-2-deoxy-D-glucose (FDG) is a useful diagnostic tool for the detection of tumours. Using dynamic FDG PET, net metabolic clearance of FDG, K, can be calculated by Gjedde-Patlak analysis of the time course of the radioactivity concentrations in tissue and arterial blood. We examined whether time-activity curves (TACs) based on arterial blood sampling could be replaced by TACs obtained from the descending aorta in dynamic PET scans of patients with liver tumours. The study was performed in two parts, using data from dynamic liver scans with arterial blood sampling in human subjects: First, data from four patients with no liver tumours and five patients with liver tumours were used as a training group. Volumes of interest were defined in the descending aorta (aorta VOIs) by four different methods. K values were calculated based on the corresponding TACs and compared with those based on TACs of the arterial blood sample radioactivity concentrations. The aorta VOI which gave K values that were in best agreement with the K values based on the arterial blood sample measurements was called the AORTA-VOI. Use of the AORTA-VOI was subsequently tested in a test group of 19 tumour patients by comparing the K values from the AORTA-VOI with the K values based on the arterial blood sample measurements. The AORTA-VOI consisted of the sum of small regions of interest (ROIs) drawn in the centre of the aorta (approximately six pixels of 2.4x2.4 mm per transaxial slice of 3.1 mm thickness) in as many transaxial slices as possible (30-40 slices). There were no statistically significant differences between the two sets of K values. The ratio of K values in tumour tissue to K values in reference tissue was 2.1-9.7:1 (mean, 5.4:1) based on the AORTA TACs, and 2.1-8.4:1 (mean, 4.6:1) based on blood sample TACs (P>0.3). We conclude that arterial blood sampling can be replaced by the present AORTA-VOI in the calculation of the net metabolic clearance of FDG in dynamic PET studies of liver tumours in human subjects. PMID- 10805114 TI - Low-dose dobutamine gated single-photon emission tomography: comparison with stress echocardiography. AB - Perfusion scintigraphy provides important information regarding the presence of viable tissue after myocardial infarction. Defects of moderate severity, however, may represent viable myocardium, necrotic tissue or a mixture of both. In this study the presence or absence of inotropic response in the infarcted area was assessed by low-dose dobutamine tetrofosmin gated single-photon emission tomography (LDD gated SPET). Results were compared with those obtained with stress echocardiography (SE). Twenty-five patients with acute myocardial infarction were studied. Gated SPET myocardial perfusion imaging was performed 60 min after the injection of technetium-99m tetrofosmin (925 MBq) at rest using a triple-headed camera equipped with focussing collimators (Cardiofocal). Two consecutive acquisitions were performed according to a "fast" gated SPET protocol (3x20 stops, 9 s/stop, 64x64 pixel matrix, zoom 1.23) with the subjects remaining in the same position. The first acquisition was obtained at rest; the second acquisition was obtained under infusion of 10 microg kg(-1) min(-1) dobutamine. The severity of regional dysfunction, wall thickening severity (WTsev), was assessed and quantified using a method based on circumferential profile analysis. SE was performed at rest and during infusion of 5 and 10 microg kg(-1) min(-1) dobutamine. Two patients could not be analysed because of disturbing gastro intestinal activity on the perfusion study. Under dobutamine 11 patients presented a significant change in WTsev (three showed normalisation, five an improvement and three a deterioration), while in 12 patients the WTsev score remained unchanged. The overall concordance between LDD gated SPET and SE was 83%. In patients with perfusion defects of moderate severity the concordance was 90% (9/10). It may be concluded that functional changes in infarcted areas induced by dobutamine can be detected with gated SPET. Good agreement was observed between LDD gated SPET and SE for the identification of inotropic reserve in infarcted areas. PMID- 10805115 TI - Effect of beta-blockade on low-dose dobutamine-induced changes in left ventricular function in healthy volunteers: assessment by gated SPET myocardial perfusion scintigraphy. AB - Viability studies are often performed in patients receiving beta-blocking agents. However, the intake of beta-blocking agents could influence the identification of viable myocardium when low-dose dobutamine is used to demonstrate inotropic reserve. The aim of this study was to quantify the effect of beta-blockade on global and regional left ventricular function in healthy volunteers using low dose dobutamine gated single-photon emission tomographic (SPET) myocardial perfusion scintigraphy. Ten subjects were studied once "on" and once "off" beta blocker therapy (metoprolol succinate, 100 mg day(-1)). On each occasion four consecutive gated SPET acquisitions (of 7 min duration) were recorded after injection of 925 MBq technetium-99m tetrofosmin on a triple-headed camera equipped with focussing (Cardiofocal) collimators. Acquisitions were made at rest (baseline 1 and 2) and 5 min after the beginning of the infusion of 5 and 10 microg kg(-1) min(-1) dobutamine. Wall thickening (WT) was quantified using a method based on circumferential profile analysis. Left ventricular ejection fraction (LVEF) was obtained using the Cedars-Sinai algorithm. Blood pressure (BP) and heart rate (HR) were recorded at the end of each acquisition. At baseline LVEF, WT and systolic BP values under beta-blockade were not significantly different from those obtained in the non-beta-blocked state. The mean HR and diastolic BP at baseline were lower under beta-blockade. Dobutamine administration (at 5 and 10 microg kg(-1) min(-1)) induced a significant increase in WT, LVEF and systolic BP in all subjects both on and off beta-blockade. The increases in WT, LVEF and systolic BP in the beta-blocked state were less pronounced but not significantly different. HR increased significantly at 10 microg kg(-1) min(-1) dobutamine without beta-blocker administration, while no increase in HR was observed in the beta-blocked state. Beta-blocker therapy in healthy subjects attenuates the inotropic and chronotropic myocardial response to low-dose dobutamine. At doses of 5 and 10 microg kg(-1) min(-1) dobutamine, however, significant increases in global and regional left ventricular function can still be measured using consecutive gated SPET myocardial perfusion scintigraphy acquisitions even under beta-blocker therapy. PMID- 10805116 TI - How frequently is the thymus seen on whole-body iodine-131 diagnostic and post treatment scans? AB - The purpose of the study was to determine how often the thymus is visualized on whole-body radioiodine scans. One hundred and seventy-five patients had 325 diagnostic scans and 200 post-treatment scans. Activity in the mediastinum possibly consistent with the thymus was seen on seven scans in six patients. Four of these were diagnostic scans (three were second follow-up scans, and the fourth, a third follow-up scan). Three post-treatment scans demonstrated mediastinal uptake. Only one patient had persistent mediastinal uptake on both a post-treatment scan and a subsequent follow-up diagnostic scan. None of these six patients were treated as a result of this finding and none has clinical or biochemical evidence of metastatic disease. PMID- 10805117 TI - The role of technetium-99m methoxyisobutylisonitrile scintigraphy in the planning of therapy and follow-up of patients with differentiated thyroid carcinoma after surgery. AB - The aim of this study was to investigate the possible role of technetium-99m methoxyisobutylisonitrile (MIBI) scan in planning post-surgical therapy and follow-up in patients with differentiated thyroid carcinoma (DTC). Four groups of DTC patients were considered: Group 1 comprised 122 patients with high serum thyroglobulin (s-Tg) levels and negative high-dose iodine-131 scan during follow up who had previously undergone total thyroidectomy and 131I treatment. Group 2 consisted of 27 patients who had previously undergone total thyroidectomy and 131I treatment but were now considered disease-free; this group was considered as controls. Group 3 comprised 49 patients studied after total thyroidectomy but prior to 131I scan. Finally, group 4 consisted of 21 patients who had previously undergone partial thyroidectomy alone. MIBI scan, neck ultrasonography (US), and s-Tg measurements during suppressive hormonal therapy (SHT) were obtained in all patients. Neck and chest computed tomography (CT) or magnetic resonance imaging (MRI) was also performed in group 1 patients. In group 1, MIBI scan and US were very sensitive in detecting cervical lymph node metastases (93.54% and 89.24%, respectively). Furthermore, MIBI scan and US played a complementary role in several patients, yielding a global sensitivity of 97.84%. In contrast, CT/MRI sensitivity for cervical lymph node metastases was very low (43.01%). MIBI scan also showed a higher sensitivity than CT/MRI in detecting mediastinal lymph node metastases (100% vs 57.89%). Regarding distant metastases, MIBI scan provided results similar to those of conventional imaging (CT, MRI, 99mTc-methylene diphosphonate bone scan). In group 2, no false-positive cases were observed with MIBI scan (100% specificity). In group 3, MIBI scan correctly identified all the 131I-positive metastatic foci, except in two patients with micronodular pulmonary metastases that were visualised with 131I scan. In contrast, both MIBI scan and US showed low sensitivity (46.15% and 61.53%, respectively) compared with 131I scan in detecting thyroid remnants. s-Tg was increased in all patients with distant metastases but only in 56% of those with lymph node metastases. Furthermore, s-Tg was increased in 21.42% of patients with thyroid remnants alone (false-positive results). In group 4, MIBI scan was the only examination capable of detecting at an early stage a mediastinal lymph node metastasis in one patient. We conclude that the integrated MIBI scan/neck US protocol: (a) can be proposed as a first-line diagnostic procedure in the follow-up of DTC patients with high s-Tg levels and negative high-dose 131I scan, and (b) may be helpful in the follow-up of DTC patients who undergo partial thyroidectomy alone. Moreover, the combined MIBI scan/neck US/s-Tg protocol appears to be highly sensitive in identifying patients with metastatic disease after total thyroidectomy and prior to 1311 scan; consequently, it may play a prognostic role in distinguishing high risk from low-risk DTC patients. However, due to the low sensitivity of MIBI scan and neck US in detecting thyroid remnants, this diagnostic approach cannot be used as a predictor of 131I scan results. Lastly, because of the high sensitivity of MIBI scan and neck US in revealing both functioning and non-functioning metastases, this integrated protocol might be helpful in the follow-up of high risk DTC patients, particularly for the early detection of lymph node metastases in patients with undetectable s-Tg during SHT. PMID- 10805118 TI - The role of renal scintigraphy and unenhanced helical computerized tomography in patients with ureterolithiasis. AB - Unenhanced helical computerized tomography (UHCT) has recently evolved as an accurate imaging modality for determination of the presence or absence of ureterolithiasis in patients with acute flank pain. Functional renal scintigraphy is considered the gold standard for urinary tract obstruction. The objective of this study was to correlate the secondary signs of urinary obstruction on UHCT with findings of functional renal scintigraphy. UHCT was performed in 30 patients admitted to the emergency room with acute flank pain. All patients had a calcified urinary stone identified on UHCT. The location of each urinary stone was classified as ureteral or in the ureterovesical junction. The presence of secondary CT signs of ureteral obstruction was determined for each patient. After oral or intravenous hydration, a technetium-99m diethylene triamine penta-acetic acid renal scan was performed in all patients within 12 h of the CT scan. Follow up delayed scintigraphic images were obtained at 2 h and 24 h in patients with evidence of ureteral obstruction. The sensitivity, specificity and predictive values of each possible combination of CT findings were determined by comparison with the scintigraphic results. The distal ureter was the most common location for a calculus on UHCT, followed in frequency by the ureterovesical junction, proximal ureter and mid-ureter. The renograms showed high-grade, unilateral obstruction in 12 patients, indeterminate scans in five patients and normal renograms in 13 patients. The sensitivities and specificities of individual CT findings ranged from 50% to 75% and from 8% to 69%, respectively. Perinephric stranding gave the highest positive predictive value (PPV) for obstruction (69% including indeterminate renograms). None of the individual CT findings showed a statistically significant correlation with scintigraphic findings. A combination of one or two positive CT findings had a PPV of only 25% for obstruction. A combination of three or four positive CT findings gave a PPV of 70% for obstruction. Our preliminary study shows that secondary CT signs of ureterolithiasis correlate poorly with the scintigraphic findings and that they do not permit evaluation of the functional status of obstructed kidneys. Even a combination of the most frequent CT findings has a low predictive value, i.e. does not allow a decision to be made as to the most suitable treatment. Therefore, renal scintigraphy should be performed in conjunction with UHCT in all patients with ureteral calculi. PMID- 10805119 TI - Cutaneous malignant melanoma: clinical aspects, imaging modalities and treatment. AB - Cutaneous melanoma is a highly malignant tumour of the melanocytes presenting characteristic metabolic and biological features. Early detection decreases mortality and morbidity and provides the best chance for optimal clinical management. Imaging techniques, including scintigraphy, have assumed an important role in detection strategies. As a functional modality, nuclear medicine offers a variety of possibilities to assist in the clinical management of malignant melanoma. This review discusses the clinical aspects and treatment of melanoma, and the imaging techniques used for its diagnosis, staging and follow-up. A survey of currently available techniques is presented. PMID- 10805120 TI - Cholescintigraphy in the diagnosis of acute acalculous cholecystitis. PMID- 10805121 TI - Thyroglobulin before ablative radioiodine therapy. PMID- 10805122 TI - Solutions to some problems of iterative reconstruction algorithms for single photon emission tomography. PMID- 10805124 TI - Engrams. PMID- 10805123 TI - Self-injection tubing system for brain SPET in epilepsy. PMID- 10805125 TI - The influence of DNA binding on the backbone dynamics of the yeast cell-cycle protein Mbp1. AB - Mbp1 is a transcription factor involved in the regulation of the cell cycle in yeast. The N-terminus of this protein contains a DNA binding domain that includes a winged helix-turn-helix motif. The C-terminal 24 residues of this domain (the 'tail') are disordered in the crystal state, but are important for DNA binding. We have measured 15N NMR relaxation rates at 11.75 and 14.1 T to determine the dynamics of the free protein and in its complex with a specific DNA duplex. The dynamics data were quantitatively analysed using both spectral density mapping and the Lipari-Szabo formalism including the effects of chemical exchange and rotational anisotropy. A detailed analysis has been made of the effect of anisotropy, exchange and experimental precision on the recovered motional parameters. The backbone NH relaxation is affected by motions on a variety of time scales from millisecond to tens of picoseconds. The relaxation data show a structured core of 100 residues corresponding to that observed in the crystal state. Within the core of the protein, two regions on either side of the putative recognition helix (helix B) show slow (ca. 0.2 ms) conformational exchange dynamics that are quenched upon DNA binding. The C-terminal 24 residues are generally more dynamic than in the core. However, in the free protein, a stretch of approximately 8 residues in the middle of the tail show relaxation behaviour similar to that in the core, indicating a structured region. NOEs between Ala 114 in this structured part of the tail and residues in the N-terminal beta strand of the core of the protein demonstrate that the tail folds back onto the core of the protein. In the complex with DNA, the structured part of the tail extends by ca. 3 residues. These data provide a framework for understanding the biochemical data on the mechanism and specificity of DNA binding. PMID- 10805126 TI - Simultaneous determination of disulphide bridge topology and three-dimensional structure using ambiguous intersulphur distance restraints: possibilities and limitations. AB - Knowledge of the native disulphide bridge topology allows the introduction of conformational restraints between remote parts of the peptide chain. This information is therefore of great importance for the successful determination of the three-dimensional structure of cysteine-rich proteins by NMR spectroscopy. In this paper we investigate the limitations of using ambiguous intersulphur restraints [Nilges, M. (1995) J. Mol. Biol., 245, 645-660] associated with NMR experimental information to determine the native disulphide bridge pattern. Using these restraints in a simulated annealing protocol we have determined the correct topology of numerous examples, including a protein with seven disulphide bridges (phospholipase A2) and a protein in which 25% of the total number of residues are cysteines (mu-conotoxin GIIIB). We have also characterised the behaviour of the method when only limited experimental data is available, and find that the proposed protocol permits disulphide bridge determination even with a small number of restraints (around 5 NOEs--including a long-range restraint--per residue). In addition, we have shown that under these conditions the use of a reduced penalty function allows the identification of misassigned NOE restraints. These results indicate that the use of ambiguous intersulphur distances with the proposed simulated annealing protocol is a general method for the determination of disulphide bridge topology, particularly interesting in the first steps of NMR study of cysteine-rich proteins. Comparison with previously proposed protocols indicates that the presented method is more reliable and the interpretation of results is straightforward. PMID- 10805127 TI - Partial NMR assignments for uniformly (13C, 15N)-enriched BPTI in the solid state. AB - We demonstrate that high-resolution multidimensional solid state NMR methods can be used to correlate many backbone and side chain chemical shifts for hydrated micro-crystalline U-13C,15N Basic Pancreatic Trypsin Inhibitor (BPTI), using a field strength of 800 MHz for protons, magic angle sample spinning rates of 20 kHz and proton decoupling field strengths of 140 kHz. Results from two homonuclear transfer methods, radio frequency driven dipolar recoupling and spin diffusion, were compared. Typical 13C peak line widths are 0.5 ppm, resulting in Calpha-Cbeta and Calpha-CO regions that exhibit many resolved peaks. Two dimensional carbon-carbon correlation spectra of BPTI have sufficient resolution to identify and correlate many of the spin systems associated with the amino acids. As a result, we have been able to assign a large number of the spin systems in this protein. The agreement between shifts measured in the solid state and those in solution is typically very good, although some shifts near the ion binding sites differ by at least 1.5 ppm. These studies were conducted with approximately 0.2 to 0.4 micromol of enriched material; the sensitivity of this method is apparently adequate for other biological systems as well. PMID- 10805129 TI - Sensitivity enhancement in (HCA)CONH experiments. AB - A novel sensitivity-enhancement technique is proposed for experiments which correlate protein backbone resonances and start with magnetization from 13Calpha 1Halpha groups. The technique is based on replenishing magnetization lost by dipole-CSA cross-correlated relaxation of the 13Calpha spin with 13Calpha steady state magnetization. The principle is demonstrated for the (HCA)CONH experiment, resulting in 1.6-fold sensitivity enhancement compared to the HN(CA)CO experiment. Furthermore, other versions of the (HCA)CONH experiment were evaluated, including a novel experiment with spin-locking of transverse 13C-1H two-spin coherence, and a cross-correlation compensated (CA)CONH experiment which starts from 13C rather than 1H magnetization. PMID- 10805128 TI - Transverse relaxation optimised spin-state selective NMR experiments for measurement of residual dipolar couplings. AB - Three transverse relaxation optimised NMR experiments (TROSY) for the measurement of scalar and dipolar couplings suitable for proteins dissolved in aqueous iso- and anisotropic solutions are described. The triple-spin-state-selective experiments yield couplings between 1HN-13Calpha, 15N-13Calpha, 1HN-13Calpha(i 1), 15N-13Calpha(i-1), 1HN-13C'(i-1), 15N-13C'(i-1) and 13C'(i-1)-13Calpha(i-1) without introducing nonessential spectral crowding compared with an ordinary two dimensional 15N-1H correlation spectrum and without requiring explicit knowledge of carbon assignments. This set of alpha/beta-J-TROSY experiments is most useful for perdeuterated proteins in studies of structure-activity relationships by NMR to observe, in addition to epitopes for ligands, also conformational changes induced by binding of ligands. PMID- 10805130 TI - Improved photo-CIDNP methods for studying protein structure and folding. AB - Two new techniques offering considerable improvements in the quality of 1H photo CIDNP spectra of proteins are demonstrated. Both focus on the problem of progressive photo-degradation of the flavin dye used to generate polarization in exposed tryptophan, tyrosine and histidine side-chains. One approach uses rapid addition and removal of protein/flavin solution between light flashes to mix the NMR sample and introduce fresh dye into the laser-irradiated region. The other involves chemical oxidation of photo-reduced flavin by the addition of hydrogen peroxide. In both cases a larger number of scans can be accumulated before the flavin is exhausted than would otherwise be possible. The techniques are demonstrated by 600 MHz CIDNP-NOESY spectroscopy of bovine holo-alpha lactalbumin, and by real-time CIDNP observation of the refolding of bovine apo alpha-lactalbumin following rapid dilution from a high concentration of chemical denaturant. PMID- 10805131 TI - A new approach for applying residual dipolar couplings as restraints in structure elucidation. AB - Residual dipolar couplings are useful global structural restraints. The dipolar couplings define the orientation of a vector with respect to the alignment tensor. Although the size of the alignment tensor can be derived from the distribution of the experimental dipolar couplings, its orientation with respect to the coordinate system of the molecule is unknown at the beginning of structure determination. This causes convergence problems in the simulated annealing process. We therefore propose a protocol that translates dipolar couplings into intervector projection angles, which are independent of the orientation of the alignment tensor with respect to the molecule. These restraints can be used during the whole simulated annealing protocol. PMID- 10805132 TI - Sequential assignment of proline-rich regions in proteins: application to modular binding domain complexes. AB - Many protein-protein interactions involve amino acid sequences containing proline rich motifs and even polyproline stretches. The lack of amide protons in such regions complicates assignment, since 1HN-based triple-resonance assignment strategies cannot be employed. Two such systems that we are currently studying include an SH2 domain from the protein Crk with a region containing 9 prolines in a 14 amino acid sequence, as well as a WW domain that interacts with a proline rich target. A modified version of the HACAN pulse scheme, originally described by Bax and co-workers [Wang et al. (1995) J. Biomol. NMR, 5, 376-382], and an experiment which correlates the intra-residue 1Halpha, 13Calpha/13Cbeta chemical shifts with the 15N shift of the subsequent residue are presented and applied to the two systems listed above, allowing sequential assignment of the molecules. PMID- 10805133 TI - Inter- and intramolecular distance measurements by solid-state MAS NMR: determination of gramicidin A channel dimer structure in hydrated phospholipid bilayers. AB - Distance constraints are an important complement to orientational constraints. While a high-resolution monomer structure of the ion channel forming polypeptide, gramicidin A, has been solved with 120 orientational constraints, the precise geometry of the dimer interface has not been characterized. Here, using both 13C and 15N labeled gramicidin A samples in hydrated phospholipid bilayers, both inter- and intramolecular distances have been measured with a recently developed simultaneous frequency and amplitude modulation (SFAM) solid-state NMR scheme. Using this approach 15N-13C1 residual dipolar couplings across a hydrogen bond as small as 20+/-2 Hz have been characterized. While such distances are on the order of 4.2+/-0.2 A, the spectroscopy is complicated by rapid global motion of the molecular structure about the bilayer normal and channel axis. Consequently, the nominal 40 Hz dipolar coupling is averaged depending on the orientation of the internuclear vector with respect to the motional axis. The intermolecular distance confirmed the previously described monomeric structure, while the intramolecular distance across the monomer-monomer interface defined this junction and confirmed the previous model of this interface. PMID- 10805134 TI - 1H, 13C and 15N backbone assignment and secondary structure of the 19 kDa diadenosine 5', 5'''-P1, P4-tetraphosphate hydrolase from Lupinus angustifolius L. PMID- 10805135 TI - Assignment of 1H, 13C and 15N resonances of the I-domain of human leukocyte function associated antigen-1. PMID- 10805136 TI - Assignments of 1H, 13C, and 15N resonances of intramolecular dimer antifreeze protein RD3. PMID- 10805137 TI - Sequence-specific 1H, 15N and 13C resonance assignments of the inhibitory prodomain of human furin. PMID- 10805138 TI - Assignment of the 1H, 13C and 15N resonances of the C-terminal EF-hands of alpha actinin in a 14 kDa complex with Z-repeat 7 of titin. PMID- 10805139 TI - Paradoxical effects of phenobarbital on mouse hepatocarcinogenesis. AB - Phenobarbital was the first tumor promoter for rodent liver to be associated with the 2-stage or initiation-promotion concept of carcinogenesis. In rats and mice preinitiated with genotoxic carcinogens, phenobarbital administration increases the number of hepatocellular tumors by approximately 5-fold despite its nongenotoxicity. However, in mice phenobarbital occasionally exhibits strong inhibitory effects on hepatocarcinogenesis initiated with the potent carcinogen diethylnitrosamine. Both positive and negative effects of phenobarbital on hepatocytic proliferation and apoptosis, which are mechanistically involved in the promotion stage of hepatocarcinogenesis, have been described. These complex outcomes of phenobarbital treatment and their effects on hepatocarcinogenesis in mice raise serious issues regarding extrapolation of experimental data from laboratory animals to human risk assessment. Recent work suggests that the paradoxical actions of phenobarbital on hepatocarcinogenesis can be understood by consideration of qualitative diversity in initiated lesions and differential responses to promotion stimulus. PMID- 10805140 TI - Chronic effects of the novel glucocorticosteroid RPR 106541 administered to beagle dogs by inhalation. AB - The preclinical safety of RPR 106541, a novel 17-thiosteroid, was evaluated in young adult and mature dogs by inhalation exposure for 26 weeks and 52 weeks, respectively. A dry powder formulation of RPR 106541 in lactose was administered to young adult dogs (approximately 6 months of age at initiation) at doses of 0 (air and placebo controls), 10, 100, or 1,000 microg/kg/d for 26 weeks. A solution-based aerosol formulation was administered to mature dogs (approximately 10 months at initiation) from a pressurized metered dose inhaler at 0 (air and placebo controls), 10, 50, and 150 microg/kg/d for 52 weeks. Clinical evidence of glucocorticosteroid-induced immunosuppression was observed by weeks 20-26 following relatively high dose exposures (100 microg/kg/d and 1,000 microg/kg/d) in young dogs receiving the dry powder formulation for 26 weeks. Classic glucocorticosteroid effects were observed, including adrenocortical atrophy, reduced bone mass with retention of epiphyseal growth plates in long bones, prominence of stromal adipose tissue in bone marrow, and atrophy of lymphoid tissues. Inhalation administration of RPR 106541 to sexually mature dogs facilitated more definitive characterization of endocrine affects of RPR 106541 as compared with administration in younger, sexually immature animals. Significant effects in female reproductive organs included absence of corpora lutea in association with atresia of vesicular follicles within the ovaries, endometrial hyperplasia, and lobular development of mammary tissue. Discordant development of mammary tissue, accumulation of secretory material within hyperplastic endometrial glands, and hypertrophy of uterine lining epithelium in absence of ovulation were consistent with a secondary progestin effect by a potent glucocorticosteroid. PMID- 10805141 TI - Abnormal cell differentiation and p21 expression of endometrial epithelial cells following developmental exposure to diethylstilbestrol (DES). AB - Gene expression relevant to abnormal cell differentiation and altered cell cycle in endometrial epithelial cells was investigated immunohistochemically in developing mouse uteri exposed neonatally to diethylstilbestrol (DES). Female CD 1 mice were given daily s.c. injections of 2 microg of DES in corn oil or were given corn oil alone (control) at 1-5 days of age and euthanatized at 5, 6, 7, 8, 15, and 22 days of age. The endometrial epithelial cells of DES-treated mice at 5 8 days of age showed enhanced staining intensity for the estrogen receptor alpha (ER alpha), whereas the stromal cells showed decreased staining reaction; the epithelial cells showed that the protein encoded by the c-fos proto-oncogene, which plays a key role in regulating diverse estrogen-related cellular differentiation patterns, was enhanced. These cells also showed increased expression of lactoferrin, a sensitive protein marker of estrogen exposure, although the staining intensity decreased after exposure ended. The stain for p21 protein, a mitotic inhibitor which suppresses cyclin-dependent kinase activity, showed frequent positively stained cells in DES-treated mice at 5-15 days of age, whereas no accumulation of p53 protein of either wild or mutant type was detected immunohistochemically in these cells. These results indicate that suppressed cell cycle activity of endometrial epithelial cells and abnormal estrogen-related differentiation at the developmental stage following neonatal DES exposure may be caused, in part, by transient altered expression of ER alpha and expression of the p21 gene, which appears to be induced by a p53-independent mechanism. PMID- 10805142 TI - Dental pulp infarction in female rats following inhalation exposure to 2 butoxyethanol. AB - Female Fischer 344 (F344)/N rats (10 per exposure group) were exposed to 2 butoxyethanol (BE) vapors (0, 31, 62.5, 125, 250, or 500 ppm 6 h/d, 5 d/wk, for 13 weeks) to characterize its prechronic toxicity. Dental lesions consisting of bilateral multifocal dental pulp thrombosis, pulp infarction, and odontoblast infarction were noted in the maxillary incisors of 3 of 4 rats from the 500-ppm group that were sacrificed when moribund during the first week of exposure. In addition, 1 rat from the 500-ppm group that was sacrificed on day 32 had similar unilateral incisor lesions but with additional findings consistent with a unilateral maxillary incisor fracture. In contrast, rats sacrificed after 13 weeks of exposure lacked dental lesions. In conclusion, BE has the potential to cause pulp thrombosis and odontoblast infarction in female rats. The apparent variability in response to BE noted in moribund sacrificed vs terminally sacrificed rats was attributed to development of tolerance to BE-induced hemolysis and subsequent incisor regeneration. PMID- 10805143 TI - Origin and distribution of potassium bromate-induced testicular and peritoneal mesotheliomas in rats. AB - Tissue sections were examined from a 2-year bioassay of male Fischer 344 rats treated with potassium bromate administered in drinking water. All animals exhibiting peritoneal mesotheliomas also had mesotheliomas of the tunica vaginalis testis mesorchium (the reverse was not true), and the correlation of these 2 types of mesotheliomas was highly significant (r2 = 0.98). Mapping of the tunica vaginalis tumors at all time points and at all bromate concentrations revealed a pattern of increasing incidence of tumor formation on the mesothelium of the tunica vaginalis testis as a function of proximity to the mesorchial ligament. Thus, the mesorchium appears to be the major mesothelial target site for potassium bromate-mediated carcinogenesis. The frequency of occurrence of mesotheliomas by location was tunica vaginalis testis (25%), mesosplenium (20%), mesentery (10%), mesojejunum/mesocolon (8%), bladder (6.5%), mesogastrium (13%), liver serosa (5%), and kidney, small intestine, and rectum (1% each). A complete cross-section of the rat testis was prepared and used to construct a complete map of the mesothelium. Any attempt to determine the role of local dose and tissue susceptibility for the purpose of interspecies risk extrapolation must take into account the complex anatomy and physiology of this region of the visceral and testicular suspensory apparatus. Improved histologic approaches are needed for adequate assessment of this delicate suspensory system. PMID- 10805144 TI - Acute injury to differentiating Clara cells in neonatal rabbits results in age related failure of bronchiolar epithelial repair. AB - Nonciliated bronchiolar (Clara) cells are progenitor cells during lung development. During differentiation, they have a heightened injury susceptibility to environmental toxicants bioactivated by cytochrome P450 monooxygenase. When neonatal rabbits are treated with the P450-mediated cytotoxicant 4-ipomeanol (IPO), abnormal bronchiolar epithelium results. This study establishes the impact of IPO cytotoxicity on 3 stages of rabbit Clara cell differentiation, early (2.5 and 5 days postnatal [DPN]), intermediate (7 and 9 DPN), and late (15 and 21 DPN), and relates the cytotoxicity to the extent of bronchiolar repair. Neonates received a single dose of IPO (5 mg/kg) and were assessed by qualitative pathology 48 hours later for injury or at 4 weeks for repair. IPO injured the 3 stages of Clara cell differentiation to the same degree; epithelium was swollen, exfoliated, and squamated. Epithelial repair differed among the 3 stages. Bronchioles of animals treated during early and intermediate stages had simple squamous and irregularly shaped cuboidal cells. Animals treated during late stages were similar to controls. Thus, differentiating Clara cells are susceptible to injury by the P450-mediated cytotoxicant IPO, but the extent of repair varies based on when the initial injury occurs. PMID- 10805145 TI - Potential target sites in peripheral tissues for excitatory neurotransmission and excitotoxicity. AB - Glutamate receptors (GluRs) are ubiquitously present in the central nervous system (CNS) as the major mediators of excitatory neurotransmission and excitotoxicity. Neural injury associated with trauma, stroke, epilepsy, and many neurodegenerative diseases such as Alzheimer's, Huntington's, and Parkinson's diseases and amyotrophic lateral sclerosis may be mediated by excessive activation of GluRs. Neurotoxicity associated with excitatory amino acids encountered in food, such as domoic acid and monosodium glutamate, has also been linked to GluRs. Less is known about GluRs outside the CNS. Recent observations suggest that several subtypes of GluRs are widely distributed in peripheral tissues. Using immunochemical and molecular techniques, the presence of GluR subtypes was demonstrated in the rat and monkey heart, with preferential distribution within the conducting system, nerve terminals, and cardiac ganglia. GluR subtypes NMDAR 1, GluR 2/3, and mGluR 2/3 are also present in kidney, liver, lung, spleen, and testis. Further investigations are needed to assess the role of these receptors in peripheral tissues and their importance in the toxicity of excitatory compounds. Therefore, food safety assessment and neurobiotechnology focusing on drugs designed to interact with GluRs should consider these tissues as potential target/effector sites. PMID- 10805146 TI - The relative protective effects of moderate dietary restriction versus dietary modification on spontaneous cardiomyopathy in male Sprague-Dawley rats. AB - The relative protective effects of modifying dietary protein, fat, fiber, and energy content vs moderate food or dietary restriction (DR) on spontaneous cardiomyopathy of Charles River male Sprague-Dawley (SD) rats was evaluated at 1 and 2 years. For 2 years, SD rats were fed Purina Rodent Chow 5002 (21.4% protein, 5.7% fat, 4.1% fiber, 3.1 kcal/g) or a modified rodent chow 5002-9 (13.6% protein, 4.6% fat, 15.7% crude fiber, 2.4 kcal/g) ad libitum (AL) or by moderate DR at approximately 65% of the caloric intake of the AL group fed the 5002 diet. Serum lipids, carcass composition, and organ weights were evaluated and hearts were qualitatively and quantitatively examined microscopically for male SD rats at 1 and 2 years. Cardiomyopathy was characterized by the colocalization of myocardial degeneration, the development of subepicardial, perivascular, subendocardial, and interstitial fibrosis, and mononuclear inflammatory cell infiltration that increased by incidence and severity in an age dependent manner from 1 to 2 years. SD rats fed the 5002 diet AL had the greatest heart weights and the most severe cardiomyopathy, with the highest myocardial fibrotic index. These parameters were relatively decreased in the AL 5002-9 diet, the DR 5002 diet, and the DR 5002-9 diet rats at 1 and 2 years. Regardless of the type of diet fed, both AL groups had the most severe cardiomyopathy by 2 years. Moderate DR allowed isocaloric comparisons of the relative effects of modified diets on survival, obesity, and heart disease. Only slight improvements in the severity and progression of spontaneous cardiomyopathy were seen by modification of the protein, fiber, fat, and energy content of the diet if fed AL. However, moderate DR with either diet was more effective than changing the diet composition in preventing and controlling the progression of cardiomyopathy in male SD rats. PMID- 10805147 TI - Expression of cytokines and proteases in mast cells in the lesion of subcapsular cell hyperplasia in mouse adrenal glands. AB - To examine the possible roles of mast cells in the pathogenesis of subcapsular cell hyperplasia (SCH) in the adrenal glands of mice, we investigated the expression of certain cytokines, including stem cell factor (SCF), tumor necrosis factor-alpha (TNF-alpha), nerve growth factor (NGF), and basic fibroblast growth factor (bFGF), and mast cell-specific proteases, such as mouse mast cell protease (mMCP)-2 and mMCP-7. The mRNAs of c-kit (SCF receptor), bFGF, TNF-alpha, mMCP-2, and mMCP-7 were expressed in both the adrenal glands and the mouse bone marrow derived mast cells (mBMMCs). Immunoreactivities for cytokines (SCF, NGF, TNF alpha) and proteases (mMCP-2, mMCP-7) were exclusively located in the mast cells in SCH lesions. The immature mBMMCs did not express the mRNAs of SCF and NGF, whereas the mast cells in the SCH lesions showed the expression of SCF and NGF. These findings suggest that SCH may provide a favorable microenvironment for functional maturation of mast cells to produce SCF and NGF, and the mast cells in SCH lesions synthesize SCF and NGF and may, in part, use them in autocrine fashion for their survival and differentiation. Therefore, mast cells may contribute to SCH pathogenesis by producing a range of multifunctional cytokines and proteases. PMID- 10805148 TI - Acute parietal and chief cell changes induced by a lethal dose of lipopolysaccharide in mouse stomach before thrombus formation. AB - The common lipopolysaccharide (LPS)-induced gastric lesions, such as erosions or ulcers, have been investigated in depth. Little is known, however, about the acute gastric lesions following a high dose of LPS. In a time-course study, ICR female mice were given a high subcutaneous dose of LPS (50 mg/kg). Mice were sacrificed at 4, 6, 12, and 24 hours after dosing and were assessed histopathologically for acute gastric lesions. The major gastric changes were seen in the fundic region and included vacuolar degeneration of parietal cells and apoptosis of chief cells. The vacuole in parietal cells was apparent as early as 4 hours postinjection (PI), and apoptosis of chief cells was apparent at 12 hours PI. Thrombus formation, in contrast, was not seen until 24 hours PI. No erosion, ulcer, or hemorrhage was seen in any gastric region in any of the treated animals at 24 hours PI. These results indicate that a subcutaneous high dose of LPS in mice causes vacuolar degeneration of parietal cells and apoptosis of chief cells before thrombus formation or subsequent ulcerative lesions. PMID- 10805149 TI - Platelet kinetics in dogs treated with a glycoprotein IIb/IIIa peptide antagonist. AB - Platelet glycoprotein IIb/IIIa (GPIIb/IIIa) receptor antagonists have been highly effective inhibitors of platelet aggregation in preclinical studies and in clinical trials. However, decreased platelet counts have been documented in preclinical studies and in some patients receiving GPIIb/IIIa antagonists. We evaluated changes in platelet kinetics and fate in dogs receiving the GPIIb/IIIa receptor antagonist RPR 109891 orally for 4 days. Dogs receiving RPR 109891 had a 22-52% decrease in platelet count with the nadirs at 3-5 days after initiation of treatment. Platelet survival time was reduced by 19%, and platelet half-life was reduced by 63%. Indium-111-labeled platelets were rapidly cleared from the blood within 1 hour after administration of RPR 109891 on treatment days 1 and 2. This clearing was associated with a sharp increase in radioactivity in spleen but not in liver or lung. Platelet clearance was markedly attenuated on treatment days 3 and 4. Platelet counts returned to baseline within 1 week after discontinuation of treatment. These data indicate that RPR 109891 causes rapid and selective sequestration of platelets in the spleen. PMID- 10805150 TI - Nedocromil sodium inhibits canine adenovirus bronchiolitis in beagle puppies. AB - Nedocromil sodium is a nonsteroidal anti-inflammatory drug used to control asthmatic attacks. Our hypothesis is that nedocromil sodium inhibits virus induced airway inflammation, a common trigger of asthma. We nebulized nedocromil sodium into beagle dogs (n = 10, mean +/- SEM ages: 149 +/- 13 days) before and after inoculation with canine adenovirus type 2 (CAV2). Control dogs (n = 10) received saline aerosols and were either infected with CAV2 (Sal/CAV2, n = 7, mean +/- SEM ages: 140 +/- 11 days) or were not infected (Sal/Sal, n = 3, ages: 143 +/- 0 days). All dogs were anesthetized with choralose (80 mg/kg i.v.), intubated, and mechanically ventilated. Pulmonary function tests and bronchoalveolar lavage (BAL) were performed using standard techniques. Pulmonary function tests revealed no significant change between the nedocromil sodium and non-nedocromil-treated groups. The percentage of infected bronchioles was quantitated as the number of inflamed airways of 40 bronchioles examined times 100 for each dog. Nedocromil-treated dogs had significantly (p < 0.05) less mucosal inflammation (mean +/- SEM, 39% +/- 5%), epithelial denudation (36% +/- 5%), and BAL neutrophilia (11 +/- 3) than did Sal/CAV2 dogs (51% +/- 6%, 57% +/- 4%, and 33% +/- 8%, respectively). We concluded that pretreatment with nedocromil sodium aerosols attenuated CAV2-induced airway inflammation in these beagle puppies. PMID- 10805151 TI - Olfactory neuron loss in adult male CD rats following subchronic inhalation exposure to hydrogen sulfide. AB - Dysosmia and anosmia are reported to occur following human exposure to hydrogen sulfide (H2S) gas. The clinical association between H2S exposure and olfactory dysfunction in humans necessitates evaluation of the nasal cavity and olfactory system in experimental animals used to study H2S toxicity. The purpose of this study was to subchronically expose 10-week-old male CD rats to relatively low concentrations of H2S and to histologically evaluate the nasal cavity for exposure-related lesions. Rats (n = 12/group) were exposed via inhalation to 0, 10, 30, or 80 ppm H2S 6 h/d and 7 d/wk for 10 weeks. Following exposure to 30 and 80 ppm H2S, a significant increase in nasal lesions limited to the olfactory mucosa was observed. The lesions, which consisted of olfactory neuron loss and basal cell hyperplasia, were multifocal, bilaterally symmetrical, and had a characteristic rostrocaudal distribution pattern. Regions of the nasal cavity affected included the dorsal medial meatus and the dorsal and medial portions of the ethmoid recess. The no observed adverse effect level for olfactory lesions in this study was 10 ppm. For perspective, the American Conference of Governmental Industrial Hygienists threshold limit value (TLV) recommendation for H2S is currently 10 ppm (proposed revision: 5 ppm), so the concentrations employed in the present study were 3 and 8 times the TLV. These findings suggest that subchronic inhalation exposure to a relatively low level of H2S (30 ppm) can result in olfactory toxicity in rats. However, because of differences in the breathing style and nasal anatomy of rats and humans, additional research is required to determine the significance of these results for human health risk assessment. PMID- 10805152 TI - Effects of 2,4,6-trinitrotoluene in a holistic environmental exposure regime on a terrestrial salamander, Ambystoma tigrinum. AB - 2,4,6-Trinitrotoluene (TNT) is a defense-related environmental contaminant present at high concentrations in soil at some military installations. Tiger salamanders (Ambystoma tigrinum, family Ambystomatidae) were exposed to TNT in a soil matrix and fed earthworms that had also been exposed to TNT via contaminated soil. Such exposure was previously shown to result in significant accumulation of both TNT and TNT metabolites by salamanders. Following 14 days of combined oral and dermal exposures, salamanders were evaluated for signs of toxicity. Control and TNT-exposed salamanders gained weight (p < 0.025). In addition, organ to body weight ratios (kidney, liver, and spleen) were not affected by treatment. The function of splenic phagocytic cells was evaluated because these cells are sensitive to certain environmental chemical exposures. Neither the chemiluminescence response (H2O2 production) nor the phagocytic capacity of such cells were different between controls and treatment groups. In like manner, no changes were seen in the peripheral hematologic parameters investigated. Histopathologic evaluations were inconclusive, yet the liver revealed the presence of heavily pigmented iron-rich phagocytes (melanomacrophages). This investigation presents a realistic approach and preliminary data for investigating the effects of xenobiotic exposure in a soil matrix on a terrestrial vertebrate. PMID- 10805153 TI - Partial hepatectomy and bile duct ligation in rainbow trout (Oncorhynchus mykiss): histologic, immunohistochemical and enzyme histochemical characterization of hepatic regeneration and biliary hyperplasia. AB - Hepatic regeneration following partial hepatectomy (PH) and biliary hyperplasia subsequent to bile duct ligation (BDL) were characterized in rainbow trout (Oncorhynchus mykiss) by light microscopy using routine and special (immunohistochemical and enzyme histochemical) stains. Both PH and BDL involved initial hypertrophy and hyperplasia of bile preductular epithelial cells (BPDECs). BPDECs are small oval cells that form junctional complexes with hepatocytes and bile ductular cells and are commonly found in hepatic tubules of teleost liver. Proliferating BPDECs transitioned through intermediate cell types before final differentiation into large basophilic hepatocytes (following PH) or biliary epithelial cells (after BDL). Normal BPDECs and hepatocytes were both negative for cytokeratin intermediate filaments in control fish when screened with the monoclonal antibody AE1/AE3. In contrast, hyperplastic BPDECs and their progeny (intermediate cells, immature hepatocytes, ductal epithelial cells) were all strongly cytokeratin positive. Cytokeratin expression was transient in newly differentiated hepatocytes (expression decreased as hepatocytes acquired characteristics consistent with full differentiation) but was permanent in biliary epithelial cells (expression was very strong in large mature ducts). BPDECs, intermediate cells, and immature ductal cells were also strongly positive for alkaline phosphatase following BDL. Chronology of histologic events and cytokeratin and enzyme expression all support the hypothesis that BPDECs possess the capacity to differentiate into either hepatocytes or biliary epithelial cells. Thus, BPDECs may be the teleost equivalent of a bipolar hepatic stem cell in mammals. PMID- 10805154 TI - "Have you seen this?" Spontaneous cutaneous vesicular disease in Yucatan minipigs. PMID- 10805155 TI - Glomerulonephritis with fibrillary deposition in a transgenic mouse carrying the human prototype c-Ha-ras gene (rasH2 mouse). AB - Glomerulonephritis was observed in a 34-week-old transgenic CB6F1 mouse carrying the human prototype c-Ha-ras gene (rasH2 mouse) from a medium-term carcinogenicity study of N-methyl-N-nitrosourea (MNU). Lesions were characterized by severe diffuse enlargement and prominent hyalinization of glomeruli. The hyaline material was positive for periodic acid-Schiff but negative for amyloid by the Congo red method. Immunohistochemically, affected glomeruli were positive for polyclonal anti-mouse IgG. Ultrastructurally, there were characteristic subendothelial and mesangial deposits composed of fibrils showing a fingerprint pattern. Lamellae were 7.5-14.3 nm in diameter and formed multilayered structures. In addition to the renal lesions, a lymphoma was observed in the thymus, with metastasis to the spleen and some lymph nodes. However, there was no glomerulonephritis in 32 other mice bearing thymic lymphomas and in more than 40 males and females given MNU in the same study. Thus, the lesions in this mouse may have been spontaneous. Glomerulonephritis was not found in more than 120 other male and female rasH2 mice in our facility. This is the first report of glomerulonephritis in a rasH2 mouse, a promising candidate for medium-term carcinogenicity risk assessment. PMID- 10805156 TI - Recent developments in animal models of human neurodegenerative diseases. PMID- 10805157 TI - Anti-L-selectin aptamers: binding characteristics, pharmacokinetic parameters, and activity against an intravascular target in vivo. AB - Therapeutic and diagnostic applications have been envisioned for aptamers, a class of oligonucleotide ligands that bind their target molecules with high affinity and specificity (Gold, J. Biol. Chem. 270, 13581-13584, 1995). To identify parameters that are important for the in vivo activity of aptamers acting on intravascular targets, we have studied binding characteristics in vitro, pharmacokinetic parameters in Sprague-Dawley rats, and inhibitory activity in a SCID mouse/human lymphocyte model of lymphocyte trafficking for both 2'F pyrimidine 2'OH purine RNA and ssDNA anti-human L-selectin aptamers. The data indicate that aptamers with low nanomolar affinity are suitable candidates for use as in vivo reagents and that nonspecific binding to vascular cells is not an issue for efficacy. As is often observed for other reagents, plasma clearance is biphasic. Both the distribution phase and the clearance rate strongly affect in vivo activity. Pharmacokinetic parameters and in vivo activity are significantly improved by conjugating aptamers to a carrier molecule, such as polyethylene glycol (PEG). Most active in vivo is 1d40, a 2'F pyrimidine 2'OH purine aptamer conjugated to 40 kDa PEG. At a dose of 5.4 nmol/kg body weight, its duration of effect (time to 50% inhibition) is 11.2 hours, and at 1 mg or 90 nmol/kg, its plasma clearance rate (CL) is 0.4 ml/min/kg. Its ED50 is estimated to be 80 pmol/kg in preinjection dose-response experiments, compared with 4 pmol/kg for the dimeric anti-L-selectin antibody DREG56. Further improvement of in vivo activity is expected from nucleotide modifications that increase resistance to nuclease digestion for aptamers where mass is not rate limiting for clearance. Because the relationship of clearance to conjugate molecular weight (MW) is not the same for all aptamers, it is advisable to determine the relationship at the outset of in vivo studies. In summary, the data suggest that properly formulated aptamers have the capacity to be effective therapeutic agents against intravascular targets. PMID- 10805158 TI - Herpes simplex virus type 2 growth and latency reactivation by cocultivation are inhibited with antisense oligonucleotides complementary to the translation initiation site of the large subunit of ribonucleotide reductase (RR1). AB - Antisense oligonucleotides complementary to the translation initiation site of the herpes simplex virus type 2 (HSV-2) large subunit of ribonucleotide reductase (RR1) were studied for their ability to inhibit RR1 expression, HSV-2 growth, and its reactivation from latently infected ganglia. The oligomers caused a significant decrease (90%-97% inhibition) in HSV-2 RR1 expression and inhibited HSV-2 growth, with IC50 and IC90 values of 0.11 and 1.0 microM, respectively. The titers of HSV-2 mutants that are respectively deleted in the PK (ICP10deltaPK) or RR (ICP10deltaRR) domains of RR1 were also significantly (500-20,000-fold) decreased, indicating that the antisense oligomers interfere with the independent contributions of the two RR1 functions (PK and RR) toward virus growth. Inhibition was sequence specific, as evidenced by the failure of a two-base mutant (RR1TImu) to inhibit protein expression and HSV-2 growth. Furthermore, the antisense oligomers inhibited HSV-2 reactivation by cocultivation of latently infected ganglia (0/8). Virus was reactivated from ganglia cultured without oligomers, in the presence of unrelated oligomers (6/8), or in the presence of the two-base mutant RR1TImu (5/8) (p < 0.007 by two-tailed Fisher exact test). HSV-2 growth was not inhibited by antisense oligonucleotides complementary to the splice junction of HSV-2 immediate-early (IE) pre-mRNA 4 and 5 (IE4,5SA) or the translation initiation site of IE mRNA 4 (IE4TI), although the respective HSV-1 specific oligomers inhibit HSV-1 growth. PMID- 10805159 TI - Inhibition of human telomerase activity by antisense phosphorothioate oligonucleotides encapsulated with the transfection reagent, FuGENE6, in HeLa cells. AB - Telomerase, a ribonucleoprotein, synthesizes telomeric repeats (TTAGGG) onto the ends of chromosomes to maintain the constant length of the telomere DNA, and its activity is detectable in approximately 85%-90% of primary human cancers. Thus, it is postulated that human telomerase might be associated with malignant tumor development and could be a highly selective target for antitumor drug design. Antisense phosphorothioate oligonucleotides (S-ODN) were investigated for their abilities to inhibit telomerase activity in the HeLa cell line. The S-ODN were designed to be complementary to nucleotides within the RNA active site of telomerase. As a transfection reagent, FuGENE6 (Boehringer Mannheim, Mannheim, Germany) was used to enhance the cellular uptake of the oligonucleotides in cell cultures. The S-ODN encapsulated with FuGENE6 clearly inhibited telomerase activity in HeLa cells and showed sequence-specific inhibition. The encapsulated S-ODN-3 with a 19-nucleotide, (nt) chain length had inhibitory effects similar to those of the 21-mer and 23-mer S-ODN sequences (S-ODN-4 and 5), but the 15-mer and 17-mer S-ODN sequences (S-ODN-1 and 2) failed to satisfactorily prevent telomerase activity. However, apoptotic HeLa cell death was not associated with telomerase inhibition. Furthermore, the encapsulated S-ODN did not appear to be cytotoxic in terms of the cell growth rate. The oligonucleotides encapsulated with the transfection reagent had enhanced cellular uptake, and cytoplasmic and nuclear localizations were observed. However, weak fluorescent signals were observed within the cytoplasms of HeLa cells treated with the free S-ODN-3. Thus, the activities of the S-ODN were effectively enhanced by using the transfection reagent. The transfection reagent, FuGENE6, may thus be a potentially useful delivery vehicle for oligonucleotide-based therapeutics and transgenes and is appropriate for use in vitro and in vivo. PMID- 10805160 TI - Highly sensitive detection of hybridization of oligonucleotides to specific sequences of nucleic acids by application of fluorescence resonance energy transfer. AB - We show a new application of fluorescence resonance energy transfer (FRET) in two stages to detect specific sequences of nucleic acids. In the first stage, two fluorescently tagged oligonucleotides hybridize with a complementary target molecule to produce FRET. The sequences of the oligonucleotides and spectral properties of fluorophores are chosen to provide a basis for an efficient energy transfer. In the next step, the specificity of hybridization is tested by competition of labeled probes with an excess of unlabeled oligonucleotides of the same sequence. The resulting emission spectra, one obtained in the excess of unlabeled donor probe and the other produced in the excess of unlabeled acceptor probe, are compared with the spectrum from the first stage to look for differences in the emission pattern of the fluorescent labels. We show that it is possible to detect the existence of specific hybrids composed of the two probes and complementary target molecule even in very unfavorable conditions, such as the presence of unhybridized probes in the final reaction mixture, secondary nonacceptor quenching of donor probe fluorescence, and strong background emission of acceptor produced by its direct excitation with a donor excitation light. PMID- 10805161 TI - Intratumoral pharmacokinetics of oligonucleotides in a tissue-isolated tumor perfusion system. AB - The intratumoral pharmacokinetics of model oligonucleotides were studied in Walker 256 tissue-isolated tumor preparations using an in situ single-pass vascular perfusion technique. A 20-mer phosphodiester (PO) oligonucleotide, its fully phosphorothioated (PS) oligonucleotide counterpart, and an 18-mer phosphorothioated oligonucleotide containing four 2'-O-methylribonucleosides at both the 3'-end and 5'-end (PS-OMe) were used. These oligonucleotides were administered to the tumor in two ways, by constant arterial infusion and by direct intratumoral injection. In the case of constant arterial infusion, the experiments were carried out using perfusate with or without 4.7% bovine serum albumin (BSA). The protein binding of PO, PS, and PS-OMe to BSA was 46%, 87%, and 94%, respectively. No marked difference was observed between the degree of accumulation of the three types of oligonucleotides in the tumor when BSA was present in the perfusate. PS and PS-OMe showed higher degrees of accumulation in tumors compared with PO when no BSA was present. These results indicate that free (i.e., protein unbound) PS-OMe and PS have superior tumor accumulation characteristics. In the intratumoral injection experiments, PS-OMe was retained longer in tumor tissue compared with PS, suggesting that it might be useful for direct local injection into solid tumors. Thus, the present study provides useful information about the basic disposition characteristics of oligonucleotides in solid tumors. PMID- 10805162 TI - Inhibition of human cancer cell growth by inducible expression of human ribonucleotide reductase antisense cDNA. AB - Ribonucleotide reductase (RR) is a rate-limiting enzyme in DNA synthesis and repair. The enzyme consists of two dissimilar subunits, M1 and M2. It is known that the M2 subunit plays a role in tumorgenicity and metastasis. In this study, we transfected human oropharyngeal KB cancer cells with human RR M1 and M2 antisense cDNA expressed by an inducible vector system. The transfectants were double-selected with hygromycin and G418. The clones, designated KB-M1AS, KB-M2AS and KB-CAT, represented transfectant clones that contained M1 antisense cDNA, M2 antisense cDNA, and a CAT reporter gene, respectively. In a colony-forming assay, colony formation for the KB-M2AS clone decreased approximately 50% when M2 antisense mRNA expression was induced by isopropylthiogalactose (IPTG). However, the KB-M1AS clone revealed no significant inhibition under IPTG induction. RR enzyme activity, as measured by 14CDP reduction assay, revealed a 30% decrease in the IPTG-induced KB-M2AS clone relative to non-IPTG-induced samples at 144 hours. As shown by Northern blot, expression of the M2 antisense mRNA showed peaks at 48 hours and 144 hours after induction by IPTG. M2 antisense mRNA expression induced by IPTG was 33-fold greater than the uninduced control at 144 hours. Western blot analysis showed that the M2 subunit protein level decreased in the KB-M2AS clone beginning at 72 hours after induction and continued to decrease to 50% of the uninduced control at 144 hours, then showed a slight recovery at 168 hours. In conclusion, M2 antisense mRNA expression by an inducible system can effectively decrease RR M2 protein expression, reduce enzyme activity, and inhibit growth. Furthermore, this approach can be employed in future antisense investigations. PMID- 10805163 TI - Phosphorothioate oligodeoxynucleotides: what is their origin and what is unique about them? AB - The development of nucleoside phosphorothioates is described in its historical context. Examples of the interaction of phosphorothioate groups, present either in oligodeoxynucleotides or in DNA, with nucleases are presented. The structural features responsible for the resistance of the phosphorothioates toward degradation by nucleases are discussed, as are the possible reasons for the high affinity interaction of phosphorothioate oligodeoxynucleotides with certain proteins. PMID- 10805164 TI - Evaluating the mechanism of action of antiproliferative antisense drugs. PMID- 10805165 TI - Gluconeogenesis in the fetus and neonate. AB - Gluconeogenesis (GNG), a key metabolic process, involves the formation of glucose and glycogen from non-glucose precursors via pyruvate. In the strict sense, it also includes the contribution of glycerol as well as recycled glucose carbon (Cori's cycle). The developmental expression of GNG in the fetus and newborn and the quantitative contribution of GNG to glucose has been extensively investigated in humans and other mammalian species. Data from studies in rodents, rabbits, and sheep fetuses show that the development of GNG is a well-orchestrated process that is regulated by the expression of specific factors involved in the transcription of the genes for specific regulating enzymes, which catalyze GNG. These transcription factors and the genes for gluconeogenic enzymes are expressed at specific time periods during development. Although the fetus has the potential for GNG, the actual formation of glucose from pyruvate is not apparent until after birth because the rate limiting enzyme phosphoenolpyruvate carboxykinase appears only after birth in the immediate newborn period. Several tracer isotope methods have been employed to quantify the contribution of GNG to glucose. Of these, the recently developed stable isotope techniques with deuterium labeled water and the mass isotopomer distribution analysis appear to be the most precise and easily applicable in human studies. The available data show that in the human newborn, GNG appears soon after birth and contributes 30% to 70% to glucose produced. Application of new molecular biology techniques, in combination with sensitive tracer isotopic methods, will allow us to identify and examine metabolic disorders that impact GNG and help develop intervention strategies. PMID- 10805166 TI - Glucose metabolism in the developing brain. AB - As in adults, glucose is the predominant cerebral energy fuel for the fetus and newborn. Studies in experimental animals and humans indicate that cerebral glucose utilization initially is low and increases with maturation with increasing regional heterogeneity. The increases in cerebral glucose utilization with advancing age occurs as a consequence of increasing functional activity and cerebral energy demands. The levels of expression of the 2 primary facilitative glucose transporter proteins in brain, GLUT1 (blood-brain barrier and glia) and GLUT3 (neuronal), display a similar maturational pattern. Alternate cerebral energy fuels, specifically the ketone bodies and lactate, can substitute for glucose, especially during hypoglycemia, thereby protecting the immature brain from potential untoward effects of hypoglycemia. Unlike adults, glucose supplementation during hypoxia-ischemia is protective in the immature brain, whereas hypoglycemia is deleterious. Accordingly, glucose plays a critical role in the developing brain, not only as the primary substrate for energy production but also to allow for normal biosynthetic processes to proceed. PMID- 10805167 TI - Newer techniques to study neonatal hypoglycemia. AB - Severe neurological sequelae may occur after symptomatic neonatal hypoglycemia. New neuroimaging techniques allow both structural and functional detection of these disturbances. The new diagnostic modalities have shown also transient structural findings associated with neonatal hypoglycemia. The prognostic value of these techniques remains still obscure. PMID- 10805168 TI - Effects of diabetic pregnancy on the fetus and newborn. AB - Diabetes in pregnancy is unique because of the diversity of problems that can affect the embryo/fetus beginning with conception. Considerable effort has been devoted to understanding the basic developmental biology from observing young embryos in vitro or in vivo. Maternal glucose control has been identified as an important event. The preponderance of evidence indicates that rigid glucose control will minimize the incidence of anomalies incurred before 9 weeks of pregnancy. Later events are related to fetal hyperinsulinemia. These include fetal macrosomia, respiratory distress syndrome, neonatal hypoglycemia, neonatal hypocalcemia, and neonatal hypomagnesemia. Control of maternal metabolism can have a significant impact on each of the above. Finally, the long-term effects of maternal diabetes are as diverse as the pathogenetic events during pregnancy. Surprisingly, there is a significant transmission rate of 2% of type I diabetes if the mother has insulin-dependent diabetic mother, whereas the rate is 6% for the father. The Diabetes in Early Pregnancy Study showed that good maternal control was associated with normal neurodevelopmental outcome. PMID- 10805169 TI - Hypoglycemia in the neonate. AB - After a brief history of the development of neonatal hypoglycemia, this review emphasizes the current approach to the anticipation, diagnosis, and management of the neonate with a low plasma glucose concentration. Current techniques for studying the neurophysiological and endocrine-metabolic effects of significant hypoglycemia provide new approaches for establishing relevant definitions of significant hypoglycemia, its prognosis, and pathogenesis. The inadequacy of glucose oxidase strips for screening, the definition of high-risk infants, new definitions for low plasma glucose concentrations, and their treatment are presented as well as the ability of the neonate to respond to significantly low glucose values. New data concerning the hereditary aspects of hyperinsulinemia (Glaser, this issue), hereditary defects in branched-chain amino acid, 3 methylglutaconic aciduria and mitochondrial betaoxidation, and degradation of fatty acids (Ozand, this issue), the role of glucose transporters (Vannucci and Vannucci, this issue), and the newer computed tomography and magnetic resonance imaging techniques (Kinnala, this issue) to study neonatal hypoglycemia are reviewed elsewhere in this issue. PMID- 10805170 TI - Hyperinsulinism of the newborn. AB - Neonatal hyperinsulinism (HI) is a clinical syndrome of pancreatic beta-cell dysfunction characterized by failure to suppress insulin secretion in the presence of hypoglycemia. Although rare, it is the most common cause for persistent hypoglycemia in the newborn period. Treatment can be extremely difficult, and partial pancreatectomy is frequently required to prevent recurrent hypoglycemia and irreversible brain damage. In the last 5 years much has been learned about the pathophysiology of this disease. In most patients, the disease is caused by recessive mutations in either of the 2 functional subunits of the beta-cell KATP channel (SUR1 or Kir6.2). Although in most families, the disease is transmitted as an autosomal recessive trait, a novel form of transmission, resulting in focal involvement of the pancreas has recently been described. Not all patients with HI have mutations in the KATP channel genes. An activating mutation in the "glucose sensor" glucokinase has recently been reported in one family with diazoxide-responsive autosomal dominant hyperinsulinemic hypoglycemia. Also, a new syndrome of hyperinsulinism associated with benign hyperammonemia was recently described and found to be caused by activating mutations in the glutamate dehydrogenase (GDH) gene (GLUD-1). Thus, the clinical syndrome of HI can be caused by mutations in 4 different genes and can be transmitted as either a recessive or a dominant trait. These findings aid in the therapeutic decision-making process and improve the accuracy and precision of genetic counseling. Despite these recent discoveries, however, the metabolic origin of the disease is still unknown in about 50% of cases. PMID- 10805171 TI - Hypoglycemia in Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome (BWS) is an overgrowth syndrome associated with macrosomia, macroglossia, abdominal wall defects, hypoglycemia in the neonatal period and embryonal cancers of infancy and early childhood. The frequency of hypoglycemia in this population is between 30% and 50%. The majority of infants with hypoglycemia will be asymptomatic and have resolution of the hypoglycemia within the first 3 days of life. Less than 5% will have hypoglycemia beyond the neonatal period requiring either continuous feeding or a partial pancreatectomy. The cause of hypoglycemia is unclear, but direct and indirect evidence supports a hyperinsulinemia as the major factor. Recent identification of the majority of genes associated with BWS in the 11p15 region and the genotype of persistent hyperinsulinemia hypoglycemia of childhood also in the 11p15 region may provide a molecular basis for hypoglycemia in BWS, particularly for the occasional patients with hypoglycemia requiring a partial pancreatectomy. Detailed genotype phenotype evaluations are needed and should provide an insight as to why patients with BWS have hypoglycemia. PMID- 10805172 TI - Hypoglycemia in association with various organic and amino acid disorders. AB - A number of organic and amino acidemias, particularly those that involve the oxidation of fatty acids, cause hypoglycemia intermittently. This may be associated with distrubances of acid base equilibrium and accumulation of lactic acid and/or ketone bodies. When such diseases are not diagnosed rapidly, they might lead to neurological crippling and, at times, death. As a group, these disorders involve more than 1 organ and their phenotypic expression may include all or a single system. The symptoms may appear soon after birth or as late as 1 year of age. Their early recognition and rapid intervention provide rewarding clinical outcome. With the recent advances in diagnostic techniques, such as the introduction of tandem mass spectrometry (MS), screening for these diseases now can be performed because rapid identification on a large scale is possible. The phenotypes, mutations involved, pathognomonic laboratory findings, prognosis, and treatment procedures available have been reviewed for major diseases. PMID- 10805173 TI - Complexation of iron(III) and iron(II) by citrate. Implications for iron speciation in blood plasma. AB - Estimates of the concentrations and identity of the predominant complexes of iron with the low-molecular-mass ligands in vivo are important to improve current understanding of the metabolism of this trace element. These estimates require a knowledge of the stability of the iron-citrate complexes. Previous studies on the equilibrium properties of the Fe(III)-citrate and Fe(II)-citrate are in disagreement. Accordingly, in this work, glass electrode potentiometric titrations have been used to re-determine the formation constants of both the Fe(III)- and Fe(II)-citrate systems at 25 degrees C in 1.00 M (Na)Cl and the reliability of these constants has been evaluated by comparing the measured and predicted redox potentials of the ternary Fe(III)-Fe(II)-citrate system. The formation constants obtained in this way were used in computer simulation models of the low-molecular-mass iron fraction in blood plasma. Redox equilibria of iron are thus included in large models of blood plasma for the first time. The results of these calculations show the predominance of Fe(II)-carbonate complexes and a significant amount of aquated Fe(II) in human blood plasma. PMID- 10805174 TI - Metallokinetic analysis of disposition of vanadyl complexes as insulin-mimetics in rats using BCM-ESR method. AB - Among vanadium's wide variety of biological functions, its insulin-mimetic effect is the most interesting and important. Recently, the vanadyl ion (+4 oxidation state of vanadium) and its complexes have been shown to normalize the blood glucose levels of streptozotocin-induced diabetic rats (STZ-rats). During our investigations to find more effective and less toxic vanadyl complexes, the vanadyl-methylpicolinate complex (VO-MPA) was found to exhibit higher insulin mimetic activity and less toxicity than other complexes, as evaluated by both in vitro and in vivo experiments. Electron spin resonance (ESR) is capable of measuring the paramagnetic species in biological samples. We have developed the in vivo blood circulation monitoring-electron spin resonance (BCM-ESR) method to analyze the ESR signals due to stable organic radicals in real time. In the present investigation, we have applied this method to elucidate the relationship between the blood glucose normalizing effect of VO-MPA and the global disposition of paramagnetic vanadyl species. This paper describes the results of vanadyl species in the circulating blood of rats following intravenous administration of vanadyl compounds. ESR spectra due to the presence of vanadyl species were obtained in the circulating blood, and their pharmacokinetic parameters were estimated using compartment models. The results indicate that vanadyl species are distributed considerably to the peripheral tissues, as estimated by BCM-ESR, and eliminated from the body through the urine, as estimated by ESR at 77 K. The exposure of vanadyl species in the blood was found to be enhanced by VO-MPA treatment. Given these results, we concluded that the pharmacokinetic character of vanadyl species is closely related with the structure and antidiabetic activity of the vanadyl compounds. PMID- 10805175 TI - Sugar interaction with metal ions. Crystal structure and FT-IR spectroscopic study of strontium galactarate mono-hydrate. AB - The crystal structure of strontium galactarate mono-hydrate, Sr2+ x C6H8O8(2-) x H2O, Mr = 313.76, monoclinic, P2(1)/c, a = 10.268(2), b = 10.333(2), c = 10.194(2) A, beta = 117.87(3) degrees, lambda(Mo K alpha) = 0.71073 A, Z = 4, Dx = 2.180 Mg m(-3), V = 956.1(3) A3, mu = 5.676 mm(-1), F(000) = 624, T = 293(2) K, R = 0.0260 for 1690 observed reflections and 145 parameters refined, has been determined. The galactarate ion is centro-symmetrical in the crystal structure, although it contains independent half-ions. The Sr2+ ion is nine-coordinated (tricapped trigonal prism) with five Sr-O bonds from carboxylic groups, and four from hydroxyl groups. The water molecule does not take part in the coordination. Six hydrogen bonds are formed, three of them related to the water molecule. The spectroscopic evidence shows that the carboxylic acid dimers of the free acid dissociate. The asymmetric stretching vibrations of the anionic COO groups in the salt are observed at 1609 and 1548, and 1581 cm(-1), assigned to a mono-dentate and a tetra-dentate coordination, respectively. The symmetric stretching vibration is located at 1397 cm(-1). The hydroxyl groups of the galactarate skeleton take part in the metal-oxygen interaction, and the hydrogen-bonding network is rearranged upon sugar metalation. PMID- 10805176 TI - The interactions of trialkyltin compounds with lysosomes from rat liver. AB - Interactions of two trialkyltin compounds with the lysosomes from a rat liver have been studied. It is shown that these compounds induce a fast alkalinisation in the matrix of energised lysosomes. The fast alkalinisation rate is similar to the one obtained with uncouplers of the oxidative phosphorylation. An identical effect has been obtained with lysosomes energised in a chloride-free medium. This supports the hypothesis that trialkyltin compounds behave not only as Cl-/OH- exchangers, but also as proton carriers in biological membranes. This result could explain the toxicity and in particular the neurotoxicity of trialkyltin compounds. PMID- 10805177 TI - Iron complexes of the cardioprotective agent dexrazoxane (ICRF-187) and its desmethyl derivative, ICRF-154: solid state structure, solution thermodynamics, and DNA cleavage activity. AB - This study investigates the solution thermodynamics of the iron complexes of dexrazoxane (ICRF-187, (+)-1,2-bis(3,5-dioxopiperazinyl-1-yl)propane), [Fe(ADR 925)](+/0), and its desmethyl derivative ICRF-154, [Fe(ICRF-247)H2O](+/0). The solid state structure of [Fe(ICRF-247)H2O]+ is also reported. [Fe(ICRF-247)H2O]Br x 0.5NaBr x H2O crystallizes in the P42(1)2 space group with Z = 4, a = 14.9851(8), b = 14.9851(8), c = 8.0825(9) A and R = 0.03(2) for 1839 reflections and exhibits a pentagonal bipyramidal geometry with a labile water molecule occupying the seventh coordination site. Potentiometric titrations (FeL = 8.5 mM, 0.1 M NaNO3, 25 degrees C) reveal stable monomeric complexes (log Kf = 18.2 +/- 0.1, [Fe(ADR-925)]+, and 17.4 +/- 0.1, [Fe(ICRF-247)H2O]+) exist in solution at relatively low pH. Upon addition of base, the iron-bound water is deprotonated; the pKa values for [Fe(ICRF-247)H2O]+ and [Fe(ADR-925)]+ are 5.63 +/- 0.07 and 5.84 +/- 0.07, respectively. At higher pH both complexes undergo mu-oxo dimerization characterized by log Kd values of 2.68 +/- 0.07 for [Fe(ICRF 247)H2O]+ and 2.23 +/- 0.07 for [Fe(ADR-925)]+. In the presence of an oxidant and reductant, both [Fe(ICRF-247)H2O]+ and [Fe(ADR-925)]+ produce hydroxyl radicals that cleave pBR322 plasmid DNA at pH 7 in a metal complex concentration-dependent manner. At low metal complex concentrations (approximately 10(-5) M) where the monomeric form predominates, cleavage by both FeICRF complexes is efficient while at higher concentrations (approximately 5 x 10(-4) M) DNA cleavage is hindered. This change in reactivity is in part accounted for by dimer formation. PMID- 10805178 TI - Interaction between NiCl2, and nucleobases, nucleosides and nucleotides. AB - The interaction between NiCl, and nucleobases, nucleosides and nucleotides has been studied by UV-Vis difference spectrophotometry, graphite furnace atomic absorption spectrophotometry, IR spectroscopy and high pressure liquid chromatography using the technique of continuous variation. The proposed structures of the complexes formed were optimised and their electronic and vibrational spectra generated using the molecular modelling program HyperChem 5. Ni2+ reacts with guanine, 2'-dGMP, GMP, adenine and AMP to form 1:1 complexes Ni(Guanine)(H2O)5, Ni(2'-dGMP)(H2O)5, Ni(GMP)(H2O)5, Ni(Adenine)(H2O)5, and Ni(AMP)(H2O)5 respectively. In these complexes, Ni2+ is believed to be bonded to the N7 atom of adenine and guanine. PMID- 10805179 TI - Effect of sodium polyacrylate on the hydrolysis of octacalcium phosphate. AB - Octacalcium phosphate (OCP) hydrolysis into hydroxyapatite (HA) has been investigated in aqueous solutions at different concentrations of sodium polyacrylate (NaPA). In the absence of the polyelectrolyte, OCP undergoes a complete transformation into HA in 48 h. The hydrolysis is inhibited by the polymer, which is significantly adsorbed on the crystals, up to about 22 wt.%. A polymer concentration of 10(-2) mM is sufficient to cause a partial inhibition of OCP to HA transformation, which is completely hindered at higher concentrations. The small platelet-like crystals in the TEM images of partially converted OCP can display electron diffraction patterns characteristic either of OCP single crystals or of polycrystalline HA, whereas the much bigger plate-like crystals exhibit diffraction patterns characteristic of OCP single crystals. The polyelectrolyte adsorption on OCP crystals is accompanied by an increase of their mean length and by a significant reduction of the coherence length of the perfect crystalline domains along the c-axis direction. It is suggested that the carboxylate-rich polyelectrolyte is adsorbed on the hydrated layer of the OCP (100) face, thus inhibiting its in situ hydrolysis into HA. PMID- 10805180 TI - Kinetics of chromium(V) formation and reduction in fronds of the duckweed Spirodela polyrhiza--a low frequency EPR study. AB - The uptake of chromate by the duckweed Spirodela polyrhiza was investigated with atomic absorption spectroscopy and the reduction of Cr(VI) to Cr(V) was measured using low frequency EPR spectroscopy. The biphasic kinetics of the uptake was fitted to parameters of a proposed kinetic model. Another model was developed to simulate chromate reduction. The first step of chromate reduction was found to be much faster than the uptake of Cr(VI) from the free space. Most probably, this step occurs already in the cell wall or on the cell membrane surface. Further reduction of Cr(V) to Cr(III) was estimated to be slower. The disappearance of the Cr(V) signal, following transfer of the plants into a Cr-free solution, lasted several tens of hours; the kinetics was mono- or biexponential depending on the length of Cr loading. The rate constants for Cr reduction in living plants were determined for the first time. PMID- 10805181 TI - How amino acids control the binding of Cu(II) ions to DNA. Part III. A novel interaction of a histidine complex with DNA. AB - L-Histidine Cu(II) complex bound to DNA showed broad EPR signals characteristic of the aggregated Cu(II) species, which could be observed even when the molar ratio of L-histidine to Cu(II) ion was smaller than unity. The signal for the DNA fibers changed with the orientation of the fibers in the static magnetic field. Based on these results, the signal was assigned to a mono-histidine Cu(II) complex stereospecifically aggregated in a groove or along a phosphodiester chain of the double helical DNA. In contrast to the L-histidine complex, the D histidine complex bound to DNA did not show such broad signals and the observed spectra for the complex on B-form DNA fibers at -150 degrees C were simulated assuming that the g1 axis of the mono-D-histidine complex tilts by about 55 degrees from the DNA-fiber axis. Addition of some deoxy-nucleotides, but not deoxy-nucleosides, to a solution of a mono-histidine complex resulted in the formation of a dinuclear ternary complex with different structures for L- or D histidine, suggesting the possibility that the stereospecific aggregation of the L-histidine complex on a double helical DNA was mediated by the phosphodiester backbones. PMID- 10805182 TI - A scanning tunnelling microscopy study of Clostridium pasteurianum rubredoxin. AB - Scanning tunnelling microscopy (STM), which can provide 'direct' and 'non averaged' information on molecular structure in three dimensions, has been used to achieve sub-molecular resolution in a 'single molecule' of rubredoxin, an important iron-sulphur protein, at the gold (111)/water interface. The metal ligand site [Fe(III)-Cys4] appears distinct because of an enhancement of the tunnelling current over this region compared to the surrounding protein structure. PMID- 10805183 TI - Is *OH the active Fenton intermediate in the oxidation of ethanol? AB - A re-examination of the data of Rush and Koppenol (J. Inorg. Biochem. 29 (1987) 199) on the competitive oxidation of C2H5OH and Fe2+ by Fenton's reagent shows that the ratio of the rate constants of the two reactions is 3.2 and not 6.3. The significance of this finding is that it is not possible to identify the active intermediate in the Fenton reaction with the *OH radical. PMID- 10805184 TI - Clinical spectrum of 500 children with neurocysticercosis and response to albendazole therapy. AB - Neurocysticercosis is a major cause of neurologic illness worldwide. Its manifestations are variable, and somewhat different when it occurs in children. Controversy exists regarding anticysticercal therapy. The clinical, laboratory, and radiographic features of 500 consecutive children with neurocysticercosis were studied; the children were then followed prospectively and their response to albendazole therapy was analyzed. Diagnosis of neurocysticercosis was based primarily on neuroimaging. Computed tomographic (CT) scans, neurocysticercosis serology, chest radiographs, and Mantoux tests were done in all children, and magnetic resonance imaging scans in 10%. All children with multiple lesions, and some randomly allocated children with single, small, enhancing CT lesions received albendazole. CT scans were repeated after 3 to 6 months. There were 272 boys and 228 girls, age range 1 6/12 to 12 6/12 years. Seizures were present in 94.8% of cases; 83.7% had focal seizures. Features of raised intracranial pressure were seen in 30% of patients and focal neurodeficit in 4%. Single lesions were seen in 76% of the children, with perilesional edema in 57.4%. Thirty-four children who had multiple cysts and received albendazole underwent serial CT evaluation. Four showed disappearance of lesions and 22 had reductions in the size or number, to give an overall improvement rate of 76%. Serial CT studies were available on 176 children with single lesions, 90 of whom received albendazole. Improvement (disappearance or reduction in the size of lesions) was observed in 91% (82 of 90) of albendazole-treated children versus 85% (73 of 86) of untreated children. This difference was not significant. No significant side effects of albendazole were reported. These data indicate that partial seizures and single parenchymal cysts are the most frequent clinical and neuroradiographic manifestations of neurocysticercosis in children. Although albendazole therapy should be considered, especially in children with multiple lesions, many children with isolated neurocysticercosis will improve without antiparasitic therapy. PMID- 10805185 TI - Evaluation of botulinum toxin A therapy in children with adductor spasm by gross motor function measure. AB - Intramuscular injection of botulinum neurotoxin A is a relatively new method for treating spastic movement disorders in children. One major goal of any therapy for patients with movement disorders is to improve gross motor function. In this study, 18 patients with adductor spasm were treated with botulinum neurotoxin A. Treatment effect was determined with the Gross Motor Function Measure, a standardized, validated instrument designed to assist in assessment of gross motor function. Spastic muscle hyperactivity and joint mobility were evaluated by the modified Ashworth Scale and by range of motion, respectively. Compared to pretreatment values, significant improvement in gross motor function (P < .010), decrease in the modified Ashworth Scale, and increase in the range of motion (P < .010) were achieved. Patients with moderate impairment of gross motor function (classed at level III and level IV in the Gross Motor Function Classification System) benefited most from treatment. In patients with severe handicap (level V), only one of five treated patients showed improvement in gross motor function. Nevertheless, all patients in this subgroup benefited from improved ease in hygienic care. In conclusion, we have demonstrated that for most children with moderate functional impairment, the Gross Motor Function Measure is a useful instrument for objective documentation of improvements of gross motor function following treatment with botulinum neurotoxin A. PMID- 10805186 TI - Childhood epilepsy with occipital paroxysms and benign nocturnal childhood occipital epilepsy. AB - Two types of childhood epilepsy have recently been reported: childhood epilepsy with occipital paroxysms, and benign nocturnal childhood occipital epilepsy. This article reports the clinical evolution, electroencephalographic (EEG) changes, and response to therapy of eight children with childhood epilepsy with occipital paroxysms (five boys and three girls, aged from 1 1/12 to 8 years) and eight children with benign nocturnal childhood occipital epilepsy (six boys and two girls, aged from 1 4/12 to 8 3/12 years). A careful clinical and EEG follow-up of at least 7 years was carried out for all patients. At the end of follow-up, all but one of the patients with childhood epilepsy with occipital paroxysms were seizure-free, and only two were still receiving anticonvulsant drugs. All but three children had a normal EEG, and normal mental development was observed in all but two cases. Patients with benign nocturnal childhood occipital epilepsy had a good long-term prognosis; all but two children with benign nocturnal childhood occipital epilepsy had a normal EEG. These two patients showed learning disabilities and poor school performances, and required remedial education. Therefore, although childhood epilepsy with occipital paroxysms and benign nocturnal childhood occipital epilepsy are two different types of epilepsy, the long-term prognosis seems to be similar. PMID- 10805187 TI - Acute pediatric rhabdomyolysis. AB - Rhabdomyolysis is a relatively common condition that may occur intermittently in chronic and inflammatory myopathy, muscular dystrophy, and metabolic myopathy. Rhabdomyolysis can also present acutely in otherwise healthy individuals. The list of etiologies for acute muscle cell lysis is enormous, with new causes described yearly. Series on acute pediatric rhabdomyolysis have not yet been published. This article describes a retrospective review of children admitted to the authors' institution during an 8-year period in whom rhabdomyolysis was recognized as a complication during their hospital stay. Patients with intermittent or relapsing rhabdomyolysis were excluded. Nineteen children were identified. Trauma (five cases), nonketotic hyperosmolar coma (two cases), viral myositis (two cases), dystonia (two cases), and malignant hyperthermia-related conditions (two cases) were the most common causes of rhabdomyolysis. Acute renal failure was the most frequent complication, occurring in 42% of cases. The mean age of renal failure patients was 13.9 years, compared to 8 years for non-renal failure children. Careful assessment of the initial urinalysis would have suggested a diagnosis of rhabdomyolysis in 9 of 16 patients tested. PMID- 10805189 TI - Headaches in patients with neurofibromatosis-1. AB - An analysis of patients followed with a diagnosis of neurofibromatosis-1 and headache was conducted. Characterization of headache type was done after chart review of 81 patients with neurofibromatosis-1 and headache. Consent was obtained for subsequent telephone interviews using a standardized questionnaire concerning the onset, characteristics, timing, triggers, and associated symptoms of the patients' headaches. Data was summarized and tabulated. Of 132 patients with neurofibromatosis-1, 81 were identified with any headache by screening history. Recurrent headaches were present in 77% of patients and in 47% of our neurofibromatosis-1 clinic population. Fifty-three of 81 patients were accessible for and agreeable to telephone interview. There were 23 male patients and 30 female patients aged 5 6/12 to 49 6/12 years, with a mean age of 20.9 years. Eighty-one percent reported having experienced recurrent headaches within the year. The majority reported onset of headache prior to the age of 10 years. Headache characteristics included the following: frequency of monthly or less, frontotemporal location, pulsating or pressing quality, and moderate severity (pain scale 4 to 5 out of 10). Headaches interfered with daily activities, had weekend occurrence, and had a duration less than 2 hours. Common headache triggers included stress, "change in weather," menstruation, fatigue, and certain foods. A high percentage of patients reported associated symptoms of nausea with or without vomiting (37%), phonophobia, photophobia, pallor, and visual scotoma. We classified 34% of the patients as having migraine (25% with aura, 9% without aura), 45% with nonmigrainous headache only, and 15% with mixed headache types (either intermittently), and 7.5% with other head pains. We conclude that patients with neurofibromatosis-1 are at greater risk for headaches than the general population. While the prevalence of both migraine and nonnigraine headache is somewhat greater than in the general population, the proportion of tension-type headache, especially in young children, is greater than expected. PMID- 10805188 TI - Predictors of independent walking in children with spastic diplegia. AB - A prospective study was carried out to identify predictors of independent walking in 31 children with either spastic diplegia or triplegia, observed from the age of 9 to 18 months (mean, 11 months) and followed for a mean period of 30 months (range, 24 to 36 months). Mean age at most recent examination was 41 months (range, 36 to 54 months). We used an 18-item scheme to chart the acquisition, from the prone position, of prelocomotor, sitting, and locomotor skills. Examinations were conducted every 6 months and videotaped according to a standardized procedure. At latest assessment 18 (58%) of the 31 children had achieved walking, 7 (23%) independently and 11 (35%) with assistance; 13 (42%) did not achieve walking. Ambulatory status was related to developmental quotient and visual acuity: all the children who became independent walkers had normal visual acuity and in 86% of cases a normal general development quotient. Moreover, we found a significant correlation between the number of gross motor skills achieved and the rate of achievement before 2 years of age and ambulatory status at 3 to 5 years of age. Ability to put weight on the hands while prone and to roll from supine to prone position by 18 months of age were significantly related to independent walking, while ability to sit without support was predictive only at around 24 months of age. PMID- 10805190 TI - Novel missense mutation in the L1 gene in a child with corpus callosum agenesis, retardation, adducted thumbs, spastic paraparesis, and hydrocephalus. AB - Corpus callosum agenesis, retardation, adducted thumbs, spastic paraparesis, and hydrocephalus (CRASH syndrome) is an X-linked recessive disorder caused by mutations in the neuronal cell adhesion molecule L1 (LICAM) gene. L1 plays a key role in axon outgrowth and pathfinding during the development of the nervous system. We describe the case of a boy from the United Arab Emirates who presented with CRASH syndrome. Scanning the L1 gene of the patient resulted in the discovery of a novel missense mutation: transition of a G (guanine) to T (thymine) at position 604 (G604-->T), which results in conversion of aspartic acid to tyrosine at position 202 (D202Y) of the L1 protein. It is very likely that the cerebral dysgenesis is due to the abnormal structure and function of L1. PMID- 10805191 TI - Concomitant bihemispheric cerebral ganglioglioma and hemangioma in an 18-month old child: case report. AB - This is a rare case of concomitant bihemispheric cerebral ganglioglioma and hemangioma in an 18-month-old child. There were two identical processes; a combination of ganglioglioma and hemangioma was located in the parieto-occipital region of both cerebral hemispheres. The child underwent diagnostic computed tomographic and magnetic resonance imaging examinations for seizures and right sided facial palsy, which showed the bilateral presence of parieto-occipital, oval, well-demarcated, subcortically located processes. A biparieto-occipital craniotomy was carried out and two well-demarcated abnormalities were observed and both were removed in toto. Two years after the operation, the boy had normal psychomotor development and remained free from seizures with no therapy required. Repeat magnetic resonance imaging showed no tumor recurrence. Gangliogliomas are relatively uncommon tumors in childhood, located in the supratentorial compartment and usually associated with seizures. Histopathologic and immunohistochemical analysis of the two processes found in our patient showed them to be a ganglioglioma and a hemangioma, confirming the dysontogenetic origin of these alterations and the association between malformations and tumor growth. The combination of ganglioglioma and hemangioma, especially bilateral presentation of two such identical processes in a young child, is very rare and was not found in the recent literature. PMID- 10805192 TI - Familial idiopathic West syndrome. AB - Two families, each with occurrence of West syndrome in two siblings, are presented. Monozygotic twins in family 1 developed infantile spasms at the age of 4 months. Two female siblings in family 2 started to have seizures at the age of 6 months, but 2 years apart. The family history; development prior to West syndrome; clinical, electroencephalographic, and neuroradiologic findings; diagnostic work-up; and treatment are described. The outcome in family 1 (follow up after 2 years) showed no conspicuous findings on physical and neurologic examination, and psychomotor development appropriate to cognitive, motor, and language developmental age in both twins. In family 2 (follow-up after 3 and 5 years), the older sister only was one standard deviation below mean in intellectual developmental age. Simultaneous occurrence of infantile spasms in both siblings from these two families but with variable clinical expression suggests there is a genetic susceptibility and variable phenotypic expression. Long-term follow-up will demonstrate whether these cases may be classified as "familial idiopathic West syndrome." PMID- 10805193 TI - Subacute sclerosing panencephalitis with atypical onset: clinical, computed tomographic, and magnetic resonance imaging correlations. AB - Clinical, computed tomographic (CT), and magnetic resonance imaging (MRI) correlations of subacute sclerosing panencephalitis with an atypical onset are presented in three children. In all three patients, the disease began similarly, with unilateral neurologic deficit followed by gnosis, praxis, and memory dysfunction corresponding to massive one-sided lesions. The first patient demonstrated right frontal-lobe syndrome and polymorphic extrapyramidal hyperkinesias; MRI showed a large high-signal lesion in the right frontal lobe, while CT was normal. The second patient displayed a disease onset with left-sided hemiparkinsonism and involuntary movements correlating to the MRI finding, ie, a massive rightsided occipitotemporoparietal subcortical lesion. An acute, stroke like episode represented the first symptom in the third child. CT visualized cerebral, mainly left-sided cortical atrophy. In all three children, CT and MRI revealed significantly progressing brain atrophy at the disease's latest stages. We discuss the role of MRI in detecting early pathologic changes in children with subacute sclerosing panencephalitis. PMID- 10805194 TI - Analgesic-induced headaches in a 17-month-old infant. AB - This is a case report of a 17-month-old infant with 8 weeks of constant headache, which resulted from the daily ingestion of analgesics. PMID- 10805195 TI - Exophytic juvenile pilocytic astrocytomas of the posterior fossa. AB - In children, posterior fossa juvenile pilocytic astrocytomas are common tumors. Radiologically, primarily extra-axial brainstem pilocytic astrocytomas are uncommon and extra-axial cerebellar pilocytic tumors are exceptional. We report two cases of such exophytic tumors, contrasting their presentation, imaging appearance, and prognosis. We also report the radiographic features of posterior fossa juvenile pilocytic astrocytomas from a 5-year review of our institutional experience. PMID- 10805196 TI - Cerebral arteritis following methylphenidate use. AB - Stroke is a well-documented complication of amphetamine abuse. Methylphenidate, chemically and pharmacologically similar to amphetamines, is widely used in the treatment of attention deficit disorder in children. The possibility of vasculitis connected to methylphenidate should not be surprising. A case is reported of stroke associated with ingestion of methylphenidate in an 8-year-old boy. Family history was negative and other causes of vasculitis were excluded. We draw your attention to the risk of using methylphenidate for a long period of time. PMID- 10805197 TI - Developmental changes in cerebrospinal fluid concentrations of monoamine-related substances revealed with a Coulochem electrode array system. AB - The relationship between age and cerebrospinal fluid concentrations of monoamine related substances, including kynurenine, were investigated using a Coulochem electrode array system, to clarify developmental changes in monoamine-related substances in the human central nervous system. In neurologically normal children, significant inverse correlations with age were observed for the cerebrospinal fluid concentrations of 5-hydroxytryptophan, 5-hydroxyindole-acetic acid, kynurenine, homovanillic acid, 3-methoxy-4-hydroxyphenyl glycol, and 3-O methyl-dopamine. The concentrations of their precursors, tryptophan and tyrosine, were not related to age. This is the first comprehensive study revealing developmental changes in monoamine-related substances including their precursors and metabolites. PMID- 10805198 TI - Pleomorphic xanthoastrocytoma derived from glioneuronal malformation in a child with intractable epilepsy. AB - Malformative lesions as well as neoplasms can cause intractable epilepsy in childhood. Even though the neoplastic nature of a lesion is evident in most cases, the distinction can be difficult in some patients. We present the case of a child with intractable epilepsy caused primarily by a glioneuronal malformation. Years after the first surgical intervention, a pleomorphic xanthoastrocytoma evolved from remnants of this lesion. This case suggests that glioneuronal malformations might be precursor lesions of pleomorphic xanthoastrocytomas. PMID- 10805199 TI - A child with neurofibromatosis-1 and a lumbar epidural arteriovenous malformation. AB - A 10-year-old child with neurofibromatosis-1 was evaluated for progressive lumbar scoliosis, back pain, and foot numbness. Magnetic resonance imaging showed several lumbar intraspinal and extraspinal masses consistent with neurofibromas. The mass at L3-L5 compressed the thecal sac and was thought to be the source of the symptoms. On operative exploration, a lumbar epidural arteriovenous malformation was found, which was removed in its entirety. The child's back pain and foot numbness resolved. Epidural arteriovenous malformations in patients with neurofibromatosis-1 are rare and have been reported only in the cervical spine. Our finding of a lumbar epidural arteriovenous malformation in a child with neurofibromatosis-1 demonstrates that vascular anomalies can be present throughout the spine of patients with neurofibromatosis-1 and should be considered in the differential diagnosis of any neurofibromatosis-1-related epidural mass. PMID- 10805200 TI - Asthma and the home environment. AB - Recent years have seen a global increase in asthma prevalence. This has coincided with modifications to the home environment resulting in changes to the indoor air quality. This article considers the links between indoor pollution and asthma. Exposure to a range of substances is examined. Airborne allergens such as those from house dust mites may be important. Pollution from particulate materials associated with combustion and smoking is discussed, as is the role of chemical vapors and gases including nitrogen dioxide, sulfur dioxide, and volatile organic compounds. The efficacy of various environmental controls to limit the impact of these pollutants is explored. PMID- 10805201 TI - Hyperthyroidism and asthma. AB - Two patients presented with new onset of thyrotoxicosis and they then developed episodic wheezing a few months afterward. The asthmatic attacks improved when the patients were rendered in a euthyroid state with treatment. The present paper discusses the relationship between two diseases, hyperthyroidism and asthma, and the underlying factors potentially contributing to their conditions. The present findings suggest that asthma may develop in a susceptible individual with hyperthyroidism and it is possible that the reactive oxygen species may be a contributory factor in exacerbating wheezing in our hyperthyroid patients. PMID- 10805202 TI - Validity of a modified version of the Marks Asthma Quality of Life Questionnaire. AB - To produce a scale useful for individual clinical decision making, the Marks Asthma Quality of Life Questionnaire (AQLQ-M) was modified to a 22-item scale using a 7-point Likert response scale, and the validity of the new instrument was assessed. Adult asthma subjects with moderate to severe disease, recruited from two hospitals in Adelaide, Australia, were surveyed at baseline (n = 293), and at 3-month follow-up (n = 234). Cronbach's alpha for the Total scale of the modified AQLQ-M (MAQLQ-M) was 0.97 and all subscale values exceeded 0.90. Test-retest reliability values for all scales were between 0.88 and 0.93. All correlations between disease reference measures were statistically significant to at least the p < 0.01 level. Stronger associations were seen with symptom and self-rating scales than for lung function, medication usage, and health service utilization measures of outcome. The MAQLQ-M showed good discriminative ability for all asthma symptom categories and for different FEV1 values. Moderate, statistically significant associations were seen between changes in MAQLQ-M scores and clinical measures. Higher baseline MAQLQ-M scores were associated with lower risks over 12 months for hospital admissions (odds ratio, OR = 0.58) and repeated emergency department visits (OR = 0.47). The MAQLQ-M is a highly valid measure of asthma related quality of life. PMID- 10805203 TI - Long-term safety of a non-chlorofluorocarbon-containing triamcinolone acetonide inhalation aerosol in patients with asthma. Azmacort HFA Study Group. AB - In response to environmental concerns regarding chlorofluorocarbon (CFC), two new triamcinolone acetonide (TAA) inhalation aerosol (Azmacort Inhalation Aerosol) formulations have been developed using a more environmentally favorable propellant, HFA-134a (1,1,1,2-tetrafluoroethane). This multicenter, open-label study evaluated the safety of switching asthma patients from TAA-CFC to one of two TAA-HFA formulations. After a 2- or 4-week baseline period during which patients received only CFC-containing TAA Inhaler, 552 patients were randomized to receive TAA-HFA 75 or 225 microg for 6 or 12 months. A total of 493 patients completed treatment. Seven patients discontinued because of adverse events and two because of ineffective asthma control. The incidence of adverse events was similar in the two treatment groups, and most events were mild to moderate in severity and were not considered related to study medication. No clinically relevant suppression of the hypophyseal-pituitary-adrenal (HPA) axis was observed. Pulmonary function tests were not adversely affected by use of either study medication, and improvements were noted in forced expiratory volume in 1 sec (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25%-75%) throughout the course of treatment. This study confirms that TAA-HFA provides effective, long-term asthma control and can safely be substituted for the currently marketed CFC-containing TAA product. PMID- 10805204 TI - Asthma patients and their partners: gender differences in the relationship between psychological distress and patient functioning. AB - This study investigated gender differences in the association between partners' psychological functioning, and patients' perceptions of health functioning and healthcare utilization. Participants were 50 couples in which one person had mild to moderate asthma. Patients and partners completed several questionnaires. Male patients reported significantly greater psychological distress and utilized fewer physician services than female patients. Furthermore, female patients' perceptions of health functioning and their physician utilization were negatively affected by their partners' psychological distress. The findings suggest a need for an interdisciplinary approach to asthma management and the possible inclusion of partners in treatment, especially for female patients. PMID- 10805205 TI - Self-efficacy, activity participation, and physical fitness of asthmatic and nonasthmatic adolescent girls. AB - The prevalence of asthma, lung self-efficacy beliefs, physical activities, and physical fitness of adolescent girls were studied in a private inner-city high school serving a multiethnic, middle-class population using a questionnaire, a test of physical fitness, and an activity log. Twenty-two percent of the 172 girls were diagnosed with asthma, and an additional 15% of the girls experienced breathing difficulties. It was found that asthmatic girls were less physically fit, reported lower self-efficacy regarding their lung functioning during vigorous activities, and participated less often in those activities than nonasthmatic girls. Perceived lung efficacy was found to predict girls' subsequent participation in physical activities significantly, even when the effects of asthma, smoking, and physical fitness were controlled statistically. PMID- 10805206 TI - Prevalence of gastroesophageal reflux in asthma. AB - A high prevalence of gastroesophageal reflux disease (GER) in asthma patients has been shown in several reports from North America and Europe. However, no data from Southern Europe are available. This paper evaluates the prevalence of abnormal reflux in asthmatics, the pattern of acid reflux when present, and the relationship between asthma and GER. Eighty-one consecutive ambulatory patients with clinically stable asthma (41 women; median age 40 years, range 17-69 years) were prospectively evaluated. All patients had a thorough digestive history; baseline pulmonary function studies, including bronchoprovocation methacholine test; and ambulatory 24-hr esophageal pH monitoring. Reflux symptoms were present in 40 patients (49%). Twelve patients had abnormal GER as defined by pH testing, giving a prevalence rate of 15% (95% confidence interval 8%-24%). The presence of acid reflux was not associated with a more severe respiratory disease. Abnormal GER seems not to be a clinically significant problem in many patients with asthma in our area. PMID- 10805207 TI - Efficacy and safety of a novel beclomethasone dipropionate dry powder inhaler (Clickhaler) for the treatment of adult asthma. Amsterdam Clinical Study Group. AB - A randomized, double-blind, double-dummy protocol was used to compare the safety and efficacy of beclomethasone dipropionate (BDP) delivered by a novel dry powder inhaler (DPI, Clickhaler) or by a pressurized metered-dose inhaler (MDI) plus spacer. There was a four-week run-in period, completed by 240 adult patients, who received BDP via an MDI. Patients with stable asthma were then randomized into a 12-week treatment period and received BDP (< or =2 mg/day via DPI or MDI). There were no significant differences in morning peak expiratory flow (PEF) (primary endpoint), evening PEF, overall daytime or nighttime symptom scores, or lung function parameters (forced expiratory volume in 1 sec, forced vital capacity) between DPI and MDI. The safety profiles were similar and patient acceptability for Clickhaler was high. In conclusion, BDP administered via Clickhaler was found to be clinically equivalent to an optimally used MDI. Patients with stable asthma currently receiving BDP via MDI may be effectively switched to treatment via Clickhaler DPI. PMID- 10805208 TI - Sick building syndrome. III. Stachybotrys chartarum. AB - Increasingly, physicians are being asked to evaluate patients with putative environmentally associated illnesses. These can include a variety of problems, including infectious illnesses (Legionnaire's disease), chemical exposure in the workplace, and sick building syndromes. The latter has been an issue particularly in asthma because of the association of mold and increased bronchial responsiveness. Recently, attention has been focused on the mold Stachybotrys in human disease. Stachybotrys was first identified more than 60 years ago following an epidemic of stomatitis, rhinitis, conjunctivitis, pancytopenia, neurologic disorders, and death in horses. Since then, Stachybotrys has been identified in several outbreaks of disease in animals. It has also attracted attention as a possible agent in idiopathic pulmonary hemorrhage in infants. Stachybotrys is a relatively uncommon fungus but has been isolated from a variety of sources, including contaminated grains, tobacco, indoor air, insulator foams, and water damaged buildings with high humidity. This fungus is particularly important because it is one of a series of fungi that produces trichothecenes mycotoxins; these mycotoxins are biologically active and can produce a variety of physiological and pathologic changes in humans and animals, including modulation of inflammation and altered alveolar surfactant phospholipid concentrations. The presence of Stachybotrys in a building does not necessarily imply a cause-and effect relationship with illness, but should alert physicians and healthcare professionals to do more vigorous environmental testing. Guidelines are presented herein for intervention measures in the maintenance of heating, ventilation, and air-conditioning systems. PMID- 10805209 TI - Liver-infiltrating T lymphocytes are attracted selectively by IFN-inducible protein-10. AB - We have demonstrated that interferon-inducible protein-10 (IP-10) is produced in hepatocytes surrounded by infiltrative mononuclear cells in chronic hepatitis. To clarify the role of IP-10 in hepatitis, we examined the chemoattractive activity of IP-10 on liver-infiltrating lymphocytes in experimental animal models of hepatitis. IP-10 was specifically induced in the livers of mice treated intravenously (i.v.) with Con A, while monocyte chemotactic protein-1 (MCP-1) showed a much lower level of induction and neither RANTES nor macrophage inflammatory protein-1alpha (MIP-1alpha) was detected. The liver-infiltrating lymphocytes in Con A-induced hepatitis were attracted only by IP-10, and not by other chemokines such as RANTES, MCP-1 and MIP-1alpha. The chemoattractive effect of IP-10 was dose-dependent and was neutralized by monoclonal antibodies to IP 10. The specific effect of IP-10 on liver-infiltrating lymphocytes was also seen on those obtained from rat livers with fulminant hepatitis induced by sequential treatment with killed Propionibacterium acnes (P. acnes) and LPS. Peripheral blood lymphocytes were slightly attracted by IP-10 as well as RANTES and MIP 1alpha, while hepatic resident lymphocytes were not. On the other hand, thioglycolate-elicited peritoneal macrophages did not respond to IP-10, although they did show a response to RANTES, MCP-1 and MIP-1alpha. These results indicated that IP-10 is a specific chemoattractant for T lymphocytes in the inflammatory liver tissues and may play a specific role in the development of hepatitis. PMID- 10805210 TI - An antibody for macrophage migration inhibitory factor suppresses tumour growth and inhibits tumour-associated angiogenesis. AB - To verify the role of macrophage migration inhibitory factor (MIF) in tumourigenesis, we examined the effect of an anti-MIF antibody on tumour growth and angiogenesis. We inoculated murine colon adenocarcinoma cell line colon 26 cells subcutaneously into the flank in BALB/c mice. After nine days, we treated tumour-bearing mice with an anti-rat MIF antibody by intraperitoneal injection on days 9, 11, 13, 15, 17, 19 and 21. We found significant inhibition of tumour growth by this treatment from day 15 to day 22. Next, we implanted a chamber filled with colon 26 cells, which only passes soluble factors, in the subcutaneous fascia of the flank, and treated mice with the anti-rat MIF antibody at days 1, 3 and 5. By histological examination at day 6, angiogenesis within the subcutaneous fascia in contact with the chamber was markedly suppressed. In vitro, we added an anti-human MIF antibody to human umbilical vein endothelial cells (HUVEC) to evaluate its effect on cell growth by measurement of [3H]thymidine incorporation. We observed that the anti-MIF antibody significantly suppressed [3H]thymidine uptake by HUVEC. These results suggest the possibility that MIF is involved in tumourigenesis via promotion of angiogenesis. PMID- 10805211 TI - Essential pathogenetic role for interferon (IFN-)gamma in concanavalin A-induced T cell-dependent hepatitis: exacerbation by exogenous IFN-gamma and prevention by IFN-gamma receptor-immunoglobulin fusion protein. AB - We have studied the effects of either exogenously-administered interferon (IFN )gamma or of a nonimmunogenic mouse IFN-gamma receptor-Immunoglobulin (IFN-gamma R-Ig) fusion protein on the development of Concanavalin (Con)A-induced hepatitis in NMRI mice. PBS-treated control mice injected with 20 mg/kg ConA developed classical serological and histological signs of hepatitis with elevation of transaminases in the blood and infiltration of the liver by mononuclear cells and neutrophils. Treating the mice with rat IFN-gamma 24 h prior to and 1 h after ConA-challenge markedly exacerbated these signs of hepatitis in a dose-dependent fashion. Moreover, mice injected with lower, non hepatitogenic, doses of ConA (10, 5 mg/kg) became fully susceptible to develop hepatitis upon similar treatment with IFN-gamma. Concordantly, ConA-induced hepatitis was abrogated by either IFN-gamma R-Ig fusion protein or anti-IFN-gamma mAb. These data provide further evidence for the central pathogenetic role of endogenous IFN-gamma in ConA-induced hepatitis and demonstrate the feasibility to prevent disease development by means of a non immunogenic IFN-gamma R-Ig fusion protein. PMID- 10805213 TI - Interleukin 18--interferon gamma inducing factor--a novel player in tumour immunotherapy? AB - The list of interleukins is growing at a steady rate. Although, it is over 8 years since the initial description of interferon gamma inducing factor (IGIF, now called IL-18), this novel cytokine is still not well characterised. However, the data were sufficient to support the testing of IL-18 in experimental tumour therapy. IL-18 is produced mainly by macrophages. Similarly to IL-1beta, IL-18 does not possess a signal sequence allowing direct secretion through the plasma membrane. Although, the exact mechanism of IL-18 secretion is not confirmed, it seems that, like IL-1beta, IGIF is processed by the cysteine proteases belonging to caspase family, especially by ICE (interleukin 1beta converting enzyme). Among the target cells responding to IL-18 are T lymphocytes and NK cells, which, under the influence of IL-18, produce substantial amounts of IFN-gamma. In this respect IL-18 seems to be even stronger than IL-12. Similarly to IL-12, IL-18 stimulates cytotoxicity of T and NK cells. Moreover, it enhances FasL-mediated cytotoxicity of CD4+ T and NK cells. A potential role of IL-18 in tumour immunotherapy is discussed in this article with special emphasis on the similarities with IL-12 and the potential mechanisms of its antitumour activity in preclinical models in mice. PMID- 10805212 TI - Interleukin 7 is a potent co-stimulator of myelin specific T cells that enhances the adoptive transfer of experimental autoimmune encephalomyelitis. AB - Interleukin 7 (IL-7), originally described as a B cell growth factor, has recently been found to play a critical role in T and B lymphocyte development and function. This study evaluated the effects of IL-7 on myelin specific T cells. IL 7 strongly enhanced proliferation of proteolipid protein (PLP) 139-151 specific T cells in association with elevated secretion of the T cell growth factor IL-2. Co stimulation with IL-7 preferentially increased the levels of pro-inflammatory cytokines secreted by PLP 139-151 specific T cells and adoptive transfer of these cells into naive recipients induced a profound enhancement of experimental autoimmune encephalomyelitis, an animal model for the human disease multiple sclerosis. These results suggest that IL-7 may be a critical co-stimulatory factor that enhances the extrathymic expansion of inflammatory T cells and may play an important role in the pathogenesis of a number of inflammatory autoimmune disorders. PMID- 10805214 TI - Effects of hyperthermia and tumour necrosis factor on inflammatory cytokine secretion and procoagulant activity in endothelial cells. AB - The application of hyperthermia (HT) and tumour necrosis factor alpha (TNF) in isolation perfusion of the limb or liver results in regression of advanced cancers confined to these regions of the body in most patients and are thought to exert anti-tumour effects primarily on tumour neovasculature. However, the individual contribution of either treatment factor on endothelial cells (EC) are not known. In this study, we investigated the in vitro effects of moderate and severe HT on human umbilical vein EC (HUVEC) with and without TNF in clinically relevant doses. HUVEC were exposed to normothermia (37 degrees C) or moderate (39 degrees C) and severe (41 degrees C) HT for 90 or 180 min with or without TNF (1 microg/ml). Cell viability, cytokine secretion (IL-6, IL-8, VEGF, ICAM-1, VCAM-1, RANTES, E-selectin, P-selectin, L-selectin, and PECAM-1), and induction of procoagulant activity as reflected in tissue factor (TF) production were assessed at the end of the treatment period and at several time points thereafter. Neither HT nor TNF exerted significant cytotoxic effects on EC at the doses and temperatures used. HT resulted in increased production of PECAM-1 with little or no additional effect when combined with TNF. TNF caused increased secretion of IL 6, IL-8, ICAM-1, and VCAM-1 with little or no additional effect from HT. Increased E-selectin and RANTES levels were observed with TNF and HT only at 24 h after treatment. HT and TNF had mainly antagonistic effects on VEGF secretion with HT causing primarily decreased production and TNF causing increased VEGF secretion under all temperatures. Most notably, there was a rapid, prolonged and synergistic peak increase in procoagulant activity when TNF and HT were used in combination compared to TNF or HT treatment alone. These results indicate that TNF and HT exert primarily independent effects on inflammatory cytokine production in EC but synergistically increase procoagulant activity as reflected in TF production. These data provide a possible mechanism for the thrombotic effects in tumour neovasculature seen following isolation perfusion with these agents and provide a rationale for their combined use in this treatment setting. PMID- 10805215 TI - Identification and characterisation of transforming growth factor beta-regulated vascular smooth muscle cell genes. AB - Transforming growth factor beta (TGFbeta) is thought to play an important role in the development and/or progression of a number of vascular disorders through its numerous effects on vascular smooth muscle cells (VSMCs). In this study we sought to identify and characterize TGFbeta-regulated VSMC genes using differential mRNA display (DD-RT-PCR) analysis of RNA isolated from TGFbeta-stimulated cultured rat aortic VSMCs. Northern blot analysis was used to demonstrate that five of 19 differentially displayed bands identified represented VSMC transcripts differentially expressed by TGFbeta. DNA sequencing revealed that three of these TGFbeta regulated genes were novel whilst the remaining two were identified through homologies to known genes. One TGFbeta upregulated transcript represented the protease cathepsin B. Since cathepsins may play a role in TGFbeta activation, an enzyme-linked immunosorbent assay (ELISA) for active TGFbeta1 was used to demonstrate an effect of cathepsin B on TGFbeta1 activation in vitro using both recombinant and human serum platelet-derived latent TGFbeta1 as substrate. These results suggest that induction of cathepsin B by TGFbeta, and its ability to activate TGFbeta1, may represent a mechanism whereby the autocrine action of TGFbeta is facilitated through expression of a protein which can process its latent form. PMID- 10805216 TI - Corticosteroids and interferons inhibit cytokine-induced production of IL-8 by human endothelial cells. AB - IL-8, secreted by endothelial cells at the site of inflammation, participates in recruitment and transmigration of leukocytes. IL-8 may also have pathophysiological consequences in inflammatory and immunological disorders. We have investigated the effect of interferons (IFNs) and glucocorticosteroids (GCs) on cytokine induced secretion and production of IL-8 by human umbilical endothelial cells (HUVEC). There was a low spontaneous secretion of IL-8 by unstimulated HUVEC which increased after 6 or 24 h of stimulation with the pro inflammatory cytokines TNF-alpha or IL-1beta. IFN-gamma as well as the GCs, Dexamethasone and Budesonide, inhibited TNF-alpha induced IL-8 secretion in a dose-dependent manner. IFNs may have a general modulating effect, since IFN-alpha also inhibited the TNF-alpha-induced IL-8 secretion. There was a slight, but significant, increase in the content of intracellular IL-8 in stimulated HUVEC. However, there was no difference between stimulation with IL-1beta or TNF-alpha alone or in combination with IFNs or GCs, whereas inhibition of IL-8 secretion with monensin increased IL-8 content suggesting that IFNs and GCs inhibit synthesis rather than secretion of IL-8. In conclusion, IFNs or GCs may be useful for inhibiting IL-8 production by endothelial cells and could thus be used for therapeutic modulation of the inflammatory response. PMID- 10805217 TI - Pharmacologic doses of interleukin 8 suppositories induce follicular maturation in rabbits. AB - Interleukin 8 (IL-8) is a neutrophil chemoattractant/activating factor that plays a role in the ovarian physiology leads to investigate the effects of IL-8 on follicular maturation. Experiments were conducted using suppositories containing 100 ng, 200 ng, 400 ng IL-8, 500 microl Witepsol-base (control), human menopausal gonadotropin (im) and conjugate of fluorescein isithiocyanate-labelled IL-8. The levels of IL-8 in ovarian fluid were also measured. Histology of ovaries treated with 200 ng IL-8 showed large antral follicles filled with follicular fluid. The theca layer was divided into an interna and an externa with large extracellular spaces. The granulosa cells were loosened and appeared to be detaching from the granulosa layer. Neutrophils were localized predominantly in the theca and medulla (P<0.0001, P<0.004), and relative collagen concentration was significantly decreased in ovaries of 200 ng IL-8 (P<0.0001) compared with controls. The IL-8 was detected in ovarian fluid after 6 h (P<0.0001), 12 h (P<0.001), and 18 h (P<0.01) compared with 0 h. Fluorescein isithiocyanate labelled IL-8 conjugate was seen in the follicular wall and endometrium. We conclude that pharmacological dosage of exogenous IL-8 exerts an effect on follicular maturation through granulocyte chemotaxis and activation. PMID- 10805218 TI - Induction of interferon gamma in human gingival fibroblasts challenged with phytohaemagglutinin. AB - Interferon gamma (IFN-gamma) is a potential immunoregulatory cytokine, which is secreted mainly by cells of immune origin. In this study, we examined the capacity of human gingival fibroblasts as non-professional immune cells to express IFN-gamma messenger RNA (mRNA) and to produce the protein. Cultures of fibroblast cells were established from gingival biopsies from three children. The expression of mRNA for IFN-gamma was studied by in situ hybridization, and the level of IFN-gamma was determined by cell-released capturing ELISA. Treatment of the cells with phytohaemagglutinin (PHA) (2.5, 5.0, and 10 microg/ml) increased the number of IFN-gamma mRNA expressing cells and the protein production at 1, 6, and 24 h. Non-stimulated cells did not reveal measurable levels of IFN-gamma mRNA or the protein. The inflammatory cytokines interleukin 1beta (IL-1beta) (100 microg/ml) and tumour necrosis factor alpha (TNFalpha) (10 ng/ml) did not affect IFN-gamma mRNA expression or protein production. Treatment of the cells with 1 microM phorbol 12-myristate-13-acetate (PMA) stimulated IFN-gamma mRNA expression but had no effect on IFN-gamma protein production. We conclude that human gingival fibroblasts not only transcribe IFN-gamma mRNA but also produce the IFN gamma protein in response to PHA. The finding that human gingival fibroblasts, produce the cytokine IFN-gamma, further support the concept that these cells take an active part in the modulation of the inflammatory and immune response in the periodontal tissue. PMID- 10805219 TI - Vascular endothelial growth factor (VEGF) and melanoma. N-acetylcysteine downregulates VEGF production in vitro. AB - Vascular endothelial growth factor (VEGF), the most potent angiogenic factor identified to date, is associated with growth and metastasis of solid tumours, including melanoma. It has been shown in vitro that melanoma cells produce raised concentrations of VEGF. We examined the VEGF concentrations in plasma of 20 patients with primary melanoma, local recurrence and metastatic melanoma. We also studied the inhibiting effect of one antioxidant, N-acetylcysteine, on VEGF production in three human melanoma cell lines. We found elevated levels of VEGF (median 205 pg ml; 95 percent confidence interval, 80-414) in metastatic melanoma, with respect to primary and locally recurrent melanoma (75 pg/ml; 95 percent confidence interval, 35-130). The health control patients had levels of 25 pg/ml (95 percent confidence interval, 10-35). Human melanoma cell lines secreted VEGF in basal conditions (550-963 +/- 125 pg/ml) and N-acetylcysteine (0.5-20 mM) significantly decreased the VEGF production in a dose-dependent manner. VEGF concentrations were found to be raised in patients with primary melanoma, local recurrence, and above all, metastatic melanoma (P=0.008). N acetylcysteine inhibits VEGF production in three human melanoma cell lines. This antioxidant might have therapeutic applications in metastatic melanoma in combination with other cytotoxic drugs. PMID- 10805220 TI - Interleukin 10 suppresses phagocytic and antihyphal activities of human neutrophils. AB - We investigated the effects of human interleukin 10 (IL-10) on the antibacterial and antifungal activities of human neutrophils (PMNs) against Staphylococcus aureus and Candida albicans. Incubation of PMNs from healthy volunteers with 20 100 ng/ml of IL-10 at 37 degrees C for 1 h suppressed phagocytosis of serum opsonized S. aureus (P=0.02) and blastoconidia of C. albicans (P<0.01). In contrast, 2-100 ng/ml of IL-10 had no effect on superoxide anion production upon stimulation with phorbol myristate acetate, N-formylmethionyl leucyl phenylalanine, C. albicans blastoconidia or pseudohyphae; neither did it significantly affect conidiocidal or bactericidal activities of PMNs. However, 20 100 ng/ml of IL-10 significantly decreased PMN-induced damage of C. albicans pseudohyphae (P=0.008). The suppression of phagocytic activity of PMNs against S. aureus and blastoconidia of C. albicans as well as the impairment of PMN-induced hyphal damage may have important implications for understanding the immunosuppressive profile of IL-10 in clinical usage. PMID- 10805221 TI - Subcutaneous administration of recombinant glycosylated interleukin 6 in patients with cancer: pharmacokinetics, pharmacodynamics and immunomodulatory effects. AB - This is the first report of the serum profile of a glycosylated recombinant form of human IL-6 (rhIL-6) administered subcutaneously (1-10 microg/kg/day) in a phase I/II trial as a thrombopoietic agent in patients with advanced cancer. The pharmacodynamic effects of IL-6 were also examined. Detailed pharmacokinetic measurements were made in four patients. Peak concentrations at 5-8 h and a median t0.5 of ca. 5 h were similar to those previously reported for non glycosylated IL-6. However, higher peak concentrations and apparent differences in effective dose levels to those previously reported with the non-glycosylated form were seen. Indications of an apparent attenuation in circulating IL-6 concentrations with continuing injections were seen in eight of 10 patients examined but anti-IL-6 antibody generation was seen in only two patients. Soluble interleukin 6 receptor concentrations generally decreased. No major changes in T cell subsets were seen but expression of CD25 and CD54 by T lymphocytes significantly increased, accompanied by marked increases in soluble CD25 (sIL-2R) and CD54 (sICAM-1). No consistent change in B cells, monocytes or NK cells were seen. No evidence for induction of TNF-alpha was found. This study demonstrates similar biological effects of glycosylated rhIL-6 to those reported for the non glycosylated form but illustrates several apparent differences which are discussed further. PMID- 10805222 TI - Neutrophils synthesize and activate TGFbeta2. AB - Human polymorphonuclear neutrophils (PMN) are regulated by soluble factors such as chemotactic peptides and cytokines. PMN themselves are capable of synthesizing and releasing various cytokines, thereby taking part in the afferent or inductive limb of the immune response. We report for the first time that TGFbeta2 can be synthesized and released by PMN obtained from peripheral blood. Reverse transcriptase polymerase chain reaction (RT-PCR) showed that preparations of PMN express mRNA for transforming growth factor (TGF)beta1 and TGFbeta2 but not for TGFbeta3. Only translation of TGFbeta2 into protein was observed applying different assays. Interestingly, TGFbeta2 in supernatants from stimulated PMN could be detected in the bioactive form. PMID- 10805223 TI - IL-12 induces SDS-stable class II alphabeta dimers in murine dendritic cells. AB - We assessed the effect of rIL-12 on the expression of class II molecules and on the ratio between SDS-stable and unstable alphabeta dimers in dendritic cells. We found that in vitro exposure of the cells to IL-12 increased their surface expression of mature class II molecules, despite a marked decline in class II biosynthesis. This effect was accompanied by a striking increase in the overall proportion of SDS-stable heterodimers. PMID- 10805224 TI - Liposome-mediated transfer of IL-1 receptor antagonist gene to dispersed islet cells does not prevent recurrence of disease in syngeneically transplanted NOD mice. AB - IL-1beta is cytotoxic to pancreatic beta-cells in vitro but its role in the vicinity of beta-cells in vivo is unknown. We explored whether liposome-mediated transfer of the interleukin 1 receptor antagonist (IL-1ra) gene to islet cells might prevent recurrence of disease in syngeneically transplanted non-obese diabetic (NOD) mice. NOD mouse islet cells were transfected using liposome mediated gene transfer with a human IL-1ra cDNA construct and transplanted two days later to prediabetic NOD mice. Graft infiltration and destruction were monitored three, five and eight days posttransplantation by histology and determination of insulin and cytokine content. IL-1ra gene transfer resulted in transient expression of IL-1ra protein in islet cells in vitro as assessed by ELISA and of IL-1ra mRNA in transplanted islets as revealed by RT-PCR. However, both control and IL-1ra transfected NOD grafts exhibited massive infiltration and loss of insulin-positive cells, paralleled by a decreased insulin content. Increased IL-1ra expression did not clearly affect other cytokine profiles (IL 1beta, IFN-gamma, IL-2), except for an increase of IL-10 on day eight. In conclusion, liposome-mediated IL-1ra gene transfer to mouse islet cells results in transient expression of IL-1ra which is, however, insufficient to confer resistance to destruction of grafted insulin-producing cells in the NOD mouse. PMID- 10805225 TI - Interferon gamma inhibits proliferation and hyaluronic acid adhesion of human malignant glioma cells in vitro. AB - Malignant gliomas are frequent and the prognosis is poor. The cytokine interferon gamma (IFN-gamma) enhances several immune phenomena and may be used in immunotherapy of tumours. Therefore we investigated the influence of IFN-gamma on human cell lines T98G, U87MG, 86HG39 and 85HG66, measuring cell viability (MTT test) and proliferation (3H-thymidine uptake). IFN-gamma markedly decreased viability and proliferation of all investigated cell lines. Expression of CD44 and adhesion to hyaluronic acid (HA) are involved in glioma invasion. Influence of IFN-gamma on these two features has also been investigated. IFN-gamma markedly decreased HA-adhesion in all three investigated cell lines, whereas CD44 expression remained uninfluenced. To summarise, IFN-gamma strongly decreased cell growth and HA-adhesion of malignant glioma cell lines in vitro. We suggest further investigations to characterise better the role of IFN-gamma as a treatment opportunity for malignant gliomas. PMID- 10805226 TI - Influence of the menstrual cycle on the LPS-induced cytokine response of monocytes. AB - We previously observed a gender difference in the cytokine response of peripheral blood monocytes (PBM). The study was performed to find out whether the gender related difference might be due to hormonal changes during the menstrual cycle in healthy premenopausal females. The release of tumour necrosis factor a (TNF alpha) was reduced in the premenopausal females during the luteal phase compared to the follicular phase. Compared to the male controls the release of TNF-alpha and of interleukin 6 (IL-6) during the luteal phase was also diminished. In premenopausal females the concentration of estradiol in plasma correlated with the release of TNF-alpha and IL-6 during the luteal phase. PMID- 10805228 TI - Soluble IL-6 receptor levels in AML patients. PMID- 10805227 TI - Dual effect of IL-4 on resistance to systemic gram-negative infection and production of TNF-alpha. AB - To determine the effect of interleukin 4 (IL-4) administration in a live sepsis model characterised by high-level production of tumour necrosis factor a (TNF alpha), mice infected systemically with lethal or sublethal inocula of Pseudomonas aeruginosa were given the recombinant cytokine at different times before infection. Improved survival and decreased TNF-alpha production were observed in lethally infected mice treated with the cytokine 1 day before challenge. In contrast, increased mortality and overproduction of TNF-alpha were observed in sublethally infected mice given IL-4 at the time of infection. PMID- 10805229 TI - Nitrogen flux and sources in the Mississippi River Basin. AB - Nitrogen from the Mississippi River Basin is believed to be at least partly responsible for the large zone of oxygen-depleted water that develops in the Gulf of Mexico each summer. Historical data show that concentrations of nitrate in the Mississippi River and some of its tributaries have increased by factors of 2 to more than 5 since the early 1900s. We have used the historical streamflow and concentration data in regression models to estimate the annual flux of nitrogen (N) to the Gulf of Mexico and to determine where the nitrogen originates within the Mississippi Basin. Results show that for 1980-1996 the mean annual total N flux to the Gulf of Mexico was 1,568,000 t/year. The flux was approximately 61% nitrate as N, 37% organic N, and 2% ammonium as N. The flux of nitrate to the Gulf has approximately tripled in the last 30 years with most of the increase occurring between 1970 and 1983. The mean annual N flux has changed little since the early 1980s, but large year-to-year variations in N flux occur because of variations in precipitation. During wet years the N flux can increase by 50% or more due to flushing of nitrate that has accumulated in the soils and unsaturated zones in the basin. The principal source areas of N are basins in southern Minnesota, Iowa, Illinois, Indiana, and Ohio that drain agricultural land. Basins in this region yield 800 to more than 3100 kg total N/km2 per year to streams, several times the N yield of basins outside this region. Assuming conservative transport of N in the Mississippi River, streams draining Iowa and Illinois contribute on average approximately 35% of the total N discharged by the Mississippi River to the Gulf of Mexico. In years with high precipitation they can contribute a larger percentage. PMID- 10805230 TI - Atmospheric nitrogen in the Mississippi River Basin--emissions, deposition and transport. AB - Atmospheric deposition of nitrogen has been cited as a major factor in the nitrogen saturation of forests in the north-eastern United States and as a contributor to the eutrophication of coastal waters, including the Gulf of Mexico near the mouth of the Mississippi River. Sources of nitrogen emissions and the resulting spatial patterns of nitrogen deposition within the Mississippi River Basin, however, have not been fully documented. An assessment of atmospheric nitrogen in the Mississippi River Basin was therefore conducted in 1998-1999 to: (1) evaluate the forms in which nitrogen is deposited from the atmosphere; (2) quantify the spatial distribution of atmospheric nitrogen deposition throughout the basin; and (3) relate locations of emission sources to spatial deposition patterns to evaluate atmospheric transport. Deposition data collected through the NADP/NTN (National Atmospheric Deposition Program/National Trends Network) and CASTNet (Clean Air Status and Trends Network) were used for this analysis. NOx Tier 1 emission data by county was obtained for 1992 from the US Environmental Protection Agency (Emissions Trends Viewer CD, 1985-1995, version 1.0, September 1996) and NH3 emissions data was derived from the 1992 Census of Agriculture (US Department of Commerce. Census of Agriculture, US Summary and County Level Data, US Department of Commerce, Bureau of the Census. Geographic Area series, 1995:1b) or the National Agricultural Statistics Service (US Department of Agriculture. National Agricultural Statistics Service Historical Data. Accessed 7/98 at URL, 1998. http://www.usda.gov/nass/pubs/hisdata++ +.htm). The highest rates of wet deposition of NO3- were in the north-eastern part of the basin, downwind of electric utility plants and urban areas, whereas the highest rates of wet deposition of NH4+ were in Iowa, near the center of intensive agricultural activities in the Midwest. The lowest rates of atmospheric nitrogen deposition were on the western (windward) side of the basin, which suggests that most of the nitrogen deposited within the basin is derived from internal sources. Atmospheric transport eastward across the basin boundary is greater for NO3- than NH4+, but a significant amount of NH4+ is likely to be transported out of the basin through the formation of (NH4)2SO4 and NH4NO3 particles--a process that greatly increases the atmospheric residence time of NH4+. This process is also a likely factor in the atmospheric transport of nitrogen from the Midwest to upland forest regions in the North-East, such as the western Adirondack region of New York, where NH4+ constitutes 38% of the total wet deposition of N. PMID- 10805231 TI - Occurrence and load of selected herbicides and metabolites in the lower Mississippi River. AB - Analyses of water samples collected from the Mississippi River at Baton Rouge, Louisiana, during 1991-1997 indicate that hundreds of metric tons of herbicides and herbicide metabolites are being discharged annually to the Gulf of Mexico. Atrazine, metolachlor, and the ethane-sulfonic acid metabolite of alachlor (alachlor ESA) were the most frequently detected herbicides and, in general, were present in the largest concentrations. Almost 80% of the annual herbicide load to the Gulf of Mexico occurred during the growing season from May to August. The concentrations and loads of alachlor in the Mississippi River decreased dramatically after 1993 in response to decreased use in the basin. In contrast, the concentrations and loads of acetochlor increased after 1994, reflecting its role as a replacement for alachlor. The peak annual herbicide load occurred in 1993, when approximately 640 metric tons (t) of atrazine, 320 t of cyanazine, 215 t of metolachlor, 53 t of simazine, and 50 t of alachlor were discharged to the Gulf of Mexico. The annual loads of atrazine and cyanazine were generally 1-2% of the amount annually applied in the Mississippi River drainage basin; the annual loads of acetochlor, alachlor, and metolachlor were generally less than 1%. Despite a reduction in atrazine use, historical data do not indicate a long-term downward trend in the atrazine load to the Gulf of Mexico. Although a relation (r2 = 0.62) exists between the atrazine load and stream discharge during May to August, variations in herbicide use and rainfall patterns within subbasins can have a large effect on herbicide loads in the Mississippi River Basin and probably explain a large part of the annual variation in atrazine load to the Gulf of Mexico. PMID- 10805233 TI - Occurrence of sulfonylurea, sulfonamide, imidazolinone, and other herbicides in rivers, reservoirs and ground water in the Midwestern United States, 1998. AB - Sulfonylurea (SU), sulfonamide (SA), and imidazolinone (IMI) herbicides are relatively new classes of chemical compounds that function by inhibiting the action of a plant enzyme, stopping plant growth, and eventually killing the plant. These compounds generally have low mammalian toxicity, but plants demonstrate a wide range in sensitivity to SUs, SAs, and IMIs with over a 10,000 fold difference in observed toxicity levels for some compounds. SUs, SAs, and IMIs are applied either pre- or post-emergence to crops commonly at 1/50th or less of the rate of other herbicides. Little is known about their occurrence, fate, or transport in surface water or ground water in the USA. To obtain information on the occurrence of SU, SA, and IMI herbicides in the Midwestern United States, 212 water samples were collected from 75 surface-water and 25 ground-water sites in 1998. These samples were analyzed for 16 SU, SA and IMI herbicides by USGS Methods Research and Development Program staff using high performance liquid chromatography/mass spectrometry. Samples were also analyzed for 47 pesticides or pesticide degradation products. At least one of the 16 SUs, SAs or IMIs was detected above the method reporting limit (MRL) of 0.01 microg/l in 83% of 130 stream samples. Imazethapyr was detected most frequently (71% of samples) followed by flumetsulam (63% of samples) and nicosulfuron (52% of samples). The sum of SU, SA and IMI concentrations exceeded 0.5 microg/l in less than 10% of stream samples. Acetochlor, alachlor, atrazine, cyanazine and metolachlor were all detected in 90% or more of 129 stream samples. The sum of the concentration of these five herbicides exceeded 50 microg/l in approximately 10% of stream samples. At least one SU, SA, or IMI herbicide was detected above the MRL in 24% of 25 ground-water samples and 86% of seven reservoir samples. PMID- 10805232 TI - Finding minimal herbicide concentrations in ground water? Try looking for their degradates. AB - Extensive research has been conducted regarding the occurrence of herbicides in the hydrologic system, their fate, and their effects on human health and the environment. Few studies, however, have considered herbicide transformation products (degradates). In this study of Iowa ground water, herbicide degradates were frequently detected. In fact, herbicide degradates were eight of the 10 most frequently detected compounds. Furthermore, a majority of a herbicide's measured concentration was in the form of its degradates--ranging from 55 to over 99%. The herbicide detection frequencies and concentrations varied significantly among the major aquifer types sampled. These differences, however, were much more pronounced when herbicide degradates were included. Aquifer types presumed to have the most rapid recharge rates (alluvial and bedrock/karst region aquifers) were those most likely to contain detectable concentrations of herbicide compounds. Two indirect estimates of ground-water age (depth of well completion and dissolved-oxygen concentration) were used to separate the sampled wells into general vulnerability classes (low, intermediate, and high). The results show that the herbicide detection frequencies and concentrations varied significantly among the vulnerability classes regardless of whether or not herbicide degradates were considered. Nevertheless, when herbicide degradates were included, the frequency of herbicide compound detection within the highest vulnerability class approached 90%, and the median total herbicide residue concentration increased over an order of magnitude, relative to the parent compounds alone, to 2 microg/l. The results from this study demonstrate that obtaining data on herbicide degradates is critical for understanding the fate of herbicides in the hydrologic system. Furthermore, the prevalence of herbicide degradates documented in this study suggests that to accurately determine the overall effect on human health and the environment of a specific herbicide its degradates should also be considered. PMID- 10805234 TI - Routine determination of sulfonylurea, imidazolinone, and sulfonamide herbicides at nanogram-per-liter concentrations by solid-phase extraction and liquid chromatography/mass spectrometry. AB - Sulfonylurea (SU), imidazolinone (IMI), and sulfonamide (SA) herbicides are new classes of low-application-rate herbicides increasingly used by farmers. Some of these herbicides affect both weed and crop species at low dosages and must be carefully used. Less is known about the effect of these compounds on non-crop plant species, but a concentration of 100 ng/l in water has been proposed as the threshold for possible plant toxicity for most of these herbicides. Hence, analytical methods must be capable of detecting SUs, IMIs, and SAs at concentrations less than 100 ng/l in ambient water samples. The authors developed a two-cartridge, solid-phase extraction method for isolating 12 SU, 3 IMI, and 1 SA herbicides by using high-performance liquid chromatography/electrospray ionization-mass spectrometry (HPLC/ESI-MS) to identify and quantify these herbicides to 10 ng/l. This method was used to analyze 196 surface- and ground water samples collected from May to August 1998 throughout the Midwestern United States, and more than 100 quality-assurance and quality-control samples. During the 16 weeks of the study, the HPLC/ESI-MS maintained excellent calibration linearity across the calibration range from 5 to 500 ng/l, with correlation coefficients of 0.9975 or greater. Continuing calibration verification standards at 100-ng/l concentration were analyzed throughout the study, and the average measured concentrations for individual herbicides ranged from 93 to 100 ng/l. Recovery of herbicides from 27 reagent-water samples spiked at 50 and 100 ng/l ranged from 39 to 92%, and averaged 73%. The standard deviation of recoveries ranged from 14 to 26%, and averaged 20%. This variability reflects multiple instruments, operators, and the use of automated and manual sample preparation. Spiked environmental water samples had similar recoveries, although for some herbicides, the sample matrix enhanced recoveries by as much as 200% greater than the spiked concentration. This matrix enhancement was sample- and compound dependent. Concentrations of herbicides in unspiked duplicate environmental samples were typically within 25% of each other. The results demonstrate the usefulness of HPLC/ESI-MS for determining low-application-rate herbicides at ambient concentrations. PMID- 10805236 TI - Analysis of selected herbicide metabolites in surface and ground water of the United States. AB - One of the primary goals of the US Geological Survey (USGS) Laboratory in Lawrence, Kansas, is to develop analytical methods for the analysis of herbicide metabolites in surface and ground water that are vital to the study of herbicide fate and degradation pathways in the environment. Methods to measure metabolite concentrations from three major classes of herbicides--triazine, chloroacetanilide and phenyl-urea--have been developed. Methods for triazine metabolite detection cover nine compounds: six compounds are detected by gas chromatography/mass spectrometry; one is detected by high-performance liquid chromatography with diode-array detection; and eight are detected by liquid chromatography/mass spectrometry. Two metabolites of the chloroacetanilide herbicides--ethane sulfonic acid and oxanilic acid--are detected by high performance liquid chromatography with diode-array detection and liquid chromatography/mass spectrometry. Alachlor ethane sulfonic acid also has been detected by solid-phase extraction and enzyme-linked immunosorbent assay. Six phenylurea metabolites are all detected by liquid chromatography/mass spectrometry; four of the six metabolites also are detected by gas chromatography/mass spectrometry. Additionally, surveys of herbicides and their metabolites in surface water, ground water, lakes, reservoirs, and rainfall have been conducted through the USGS laboratory in Lawrence. These surveys have been useful in determining herbicide and metabolite occurrence and temporal distribution and have shown that metabolites may be useful in evaluation of non point-source contamination. PMID- 10805235 TI - Determination of chloroacetanilide herbicide metabolites in water using high performance liquid chromatography-diode array detection and high-performance liquid chromatography/mass spectrometry. AB - Analytical methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and high-performance liquid chromatography/mass spectrometry (HPLC/MS) were developed for the analysis of the following chloroacetanilide herbicide metabolites in water: alachlor ethanesulfonic acid (ESA); alachlor oxanilic acid; acetochlor ESA; acetochlor oxanilic acid; metolachlor ESA; and metolachlor oxanilic acid. Good precision and accuracy were demonstrated for both the HPLC-DAD and HPLC/MS methods in reagent water, surface water, and ground water. The average HPLC-DAD recoveries of the chloroacetanilide herbicide metabolites from water samples spiked at 0.25, 0.5 and 2.0 microg/l ranged from 84 to 112%, with relative standard deviations of 18% or less. The average HPLC/MS recoveries of the metabolites from water samples spiked at 0.05, 0.2 and 2.0 microg/l ranged from 81 to 118%, with relative standard deviations of 20% or less. The limit of quantitation (LOQ) for all metabolites using the HPLC-DAD method was 0.20 microg/l, whereas the LOQ using the HPLC/MS method was at 0.05 microg/l. These metabolite-determination methods are valuable for acquiring information about water quality and the fate and transport of the parent chloroacetanilide herbicides in water. PMID- 10805237 TI - Detection of persistent organic pollutants in the Mississippi Delta using semipermeable membrane devices. AB - From semipermeable membrane devices (SPMDs) placed in five Mississippi Delta streams in 1996 and 1997, the persistent organic pollutants (POPs) aldrin, chlordane, DCPA, DDT, dieldrin, endrin, heptachlor, mirex, nonachlor, and toxaphene were detected. In addition, the insecticides chlorpyriphos, endosulfan, and hexachlorocyclohexanes were detected. Two low-solubility herbicides not detected commonly in surface water, pendimethalin and trifluralin, were also detected. PMID- 10805238 TI - Use of radioimmunoassay as a screen for antibiotics in confined animal feeding operations and confirmation by liquid chromatography/mass spectrometry. AB - Approximately one-half of the 50,000,000 lb of antibiotics produced in the USA are used in agriculture. Because of the intensive use of antibiotics in the management of confined livestock operations, the potential exists for the transport of these compounds and their metabolites into our nation's water resources. A commercially available radioimmunoassay method, developed as a screen for tetracycline antibiotics in serum, urine, milk, and tissue, was adapted to analyze water samples at a detection level of approximately 1.0 ppb and a semiquantitative analytical range of 1-20 ppb. Liquid waste samples were obtained from 13 hog lagoons in three states and 52 surface- and ground-water samples were obtained primarily from areas associated with intensive swine and poultry production in seven states. These samples were screened for the tetracycline antibiotics by using the modified radioimmunoassay screening method. The radioimmunoassay tests yielded positive results for tetracycline antibiotics in samples from all 13 of the hog lagoons. Dilutions of 10-100-fold of the hog lagoon samples indicated that tetracycline antibiotic concentrations ranged from approximately 5 to several hundred parts per billion in liquid hog lagoon waste. Of the 52 surface- and ground-water samples collected all but two tested negative and these two samples contained tetracycline antibiotic concentrations less than 1 ppb. A new liquid chromatography/mass spectrometry method was used to confirm the radioimmunoassay results in 9 samples and also to identify the tetracycline antibiotics to which the radioimmunoassay test was responding. The new liquid chromatography/mass spectrometry method with online solid-phase extraction and a detection level of 0.5 microg/l confirmed the presence of chlorotetracycline in the hog lagoon samples and in one of the surface-water samples. The concentrations calculated from the radioimmunoassay were a factor of 1-5 times less than those calculated by the liquid chromatography/mass spectrometry concentrations for chlorotetracycline. PMID- 10805239 TI - Occurrence of cotton herbicides and insecticides in playa lakes of the High Plains of West Texas. AB - During the summer of 1997, water samples were collected and analyzed for pesticides from 32 playa lakes of the High Plains that receive drainage from both cotton and corn agriculture in West Texas. The major cotton herbicides detected in the water samples were diuron, fluometuron, metolachlor, norflurazon, and prometryn. Atrazine and propazine, corn and sorghum herbicides, were also routinely detected in samples from the playa lakes. Furthermore, the metabolites of all the herbicides studied were found in the playa lake samples. In some cases, the concentration of metabolites was equal to or exceeded the concentration of the parent compound. The types of metabolites detected suggested that the parent compounds had been transported to and had undergone degradation in the playa lakes. The types of metabolites and the ratio of metabolites to parent compounds may be useful in indicating the time that the herbicides were transported to the playa lakes. The median concentration of total herbicides was 7.2 microg/l, with the largest total concentrations exceeding 30 microg/l. Organophosphate insecticides were detected in only one water sample. Further work will improve the understanding of the fate of these compounds in the playa lake area. PMID- 10805240 TI - Pesticides in the atmosphere of the Mississippi River Valley, part I--rain. AB - Weekly composite rainfall samples were collected in three paired urban and agricultural regions of the Midwestern United States and along the Mississippi River during April-September 1995. The paired sampling sites were located in Mississippi, Iowa, and Minnesota. A background site, removed from dense urban and agriculture areas, was located near Lake Superior in Michigan. Herbicides were the predominant type of pesticide detected at every site. Each sample was analyzed for 47 compounds and 23 of 26 herbicides, 13 of 18 insecticides, and three of three related transformation products were detected in one or more sample from each paired site. The detection frequency of herbicides and insecticides were nearly equivalent at the paired Iowa and Minnesota sites. In Mississippi, herbicides were detected more frequently at the agricultural site and insecticides were detected more frequently at the urban site. The highest total wet depositional amounts (microg pesticide/m2 per season) occurred at the agricultural sites in Mississippi (1980 microg/m2) and Iowa (490 microg/m2) and at the urban site in Iowa (696 microg/m2). Herbicides accounted for the majority of the wet depositional loading at the Iowa and Minnesota sites, but methyl parathion (1740 microg/m2) was the dominant compound contributing to the total loading at the agricultural site in Mississippi. Atrazine, CIAT (a transformation product of atrazine and propazine) and dacthal were detected most frequently (76, 53, and 53%, respectively) at the background site indicating their propensity for long-range atmospheric transport. PMID- 10805241 TI - Pesticides in the atmosphere of the Mississippi River Valley, part II--air. AB - Weekly composite air samples were collected from early April through to mid September 1995 at three paired urban and agricultural sites along the Mississippi River region of the Midwestern United States. The paired sampling sites were located in Mississippi, Iowa, and Minnesota. A background site, removed from dense urban and agricultural areas, was located on the shore of Lake Superior in Michigan. Each sample was analyzed for 49 compounds; of these, 21 of 26 herbicides, 13 of 19 insecticides, and 4 of 4 related transformation products were detected during the study, with most pesticides detected in more than one sample. The maximum number of pesticides detected in an air sample was 18. Herbicides were the predominant type of pesticide detected at every site. Detection frequencies of most herbicides were similar at the urban and agricultural sites in Iowa and Minnesota. In Mississippi, herbicides generally were detected more frequently at the agricultural site. The insecticides chlorpyrifos, diazinon, and carbaryl, which are used in agricultural and non agricultural settings, were detected more frequently in urban sites than agricultural sites in Mississippi and Iowa. Methyl parathion was detected in 70% of the samples from the Mississippi agricultural site and at the highest concentration (62 ng/m3 air) of any insecticide measured in the study. At the background site, dacthal (100%), atrazine (35%), cyanazine (22%), and the (primarily atrazine) triazine transformation products CIAT (35%) and CEAT (17%) were detected most frequently, suggesting their potential for long-range atmospheric transport. PMID- 10805242 TI - Occurrence of pesticides in rain and air in urban and agricultural areas of Mississippi, April-September 1995. AB - In April 1995, the US Geological Survey began a study to determine the occurrence and temporal distribution of 49 pesticides and pesticide metabolites in air and rain samples from an urban and an agricultural sampling site in Mississippi. The study was a joint effort between the National Water-Quality Assessment and the Toxic Substances Programs and was part of a larger study examining the occurrence and temporal distribution of pesticides in air and rain in the Mississippi River basin. Concurrent high-volume air and wet-only deposition samples were collected weekly. The air samplers consisted of a glass-fiber filter to collect particles and tandem polyurethane foam plugs to collect gas-phase pesticides. Every rain and air sample collected from the urban and agricultural sites had detectable levels of multiple pesticides. The magnitude of the total concentration was 5-10 times higher at the agricultural site as compared to the urban site. The pesticide with the highest concentration in rain at both sites was methyl parathion. The pesticide with the highest concentration in the air samples from the agricultural site was also methyl parathion, but from the urban site the highest concentration was diazinon followed closely by chlorpyrifos. More than two decades since p,p'-DDT was banned from use in the United States, p,p'-DDE, a metabolite of p,p'-DDT, was detected in every air sample collected from the agricultural site and in more than half of the air samples from the urban site. PMID- 10805243 TI - Herbicides and herbicide degradates in shallow groundwater and the Cedar River near a municipal well field, Cedar Rapids, Iowa. AB - Water samples were collected near a Cedar Rapids, Iowa municipal well field from June 1998 to August 1998 and analyzed for selected triazine and acetanilide herbicides and degradates. The purpose of the study was to evaluate the occurrence of herbicides and herbicide degradates in the well field during a period following springtime application of herbicides to upstream cropland. The well field is in an alluvial aquifer adjacent to the Cedar River. Parent herbicide concentrations generally were greatest in June, and decreased in July and August. Atrazine was most frequently detected and occurred at the greatest concentrations; acetochlor, cyanazine and metolachlor also were detected, but at lesser concentrations than atrazine. Triazine degradate concentrations were relatively small (< 0.50 microg/l) and generally decreased from June to August. Although the rate of groundwater movement is relatively fast (approx. 1 m per day) in the alluvial aquifer near the Cedar River, deethylatrazine (DEA) to atrazine ratios in groundwater samples collected near the Cedar River indicate that atrazine and DEA probably are gradually transported into the alluvial aquifer from the Cedar River. Deisopropylatrazine (DIA) to DEA ratios in water samples indicate most DIA in the Cedar River and alluvial aquifer is produced by atrazine degradation, although some could be from cyanazine degradation. Acetanilide degradates were detected more frequently and at greater concentrations than their corresponding parent herbicides. Ethanesulfonic-acid (ESA) degradates comprised at least 80% of the total acetanilide-degradate concentrations in samples collected from the Cedar River and alluvial aquifer in June, July and August; oxanilic acid degradates comprised less than 20% of the total concentrations. ESA-degradate concentrations generally were smallest in June and greater in July and August. Acetanilide degradate concentrations in groundwater adjacent to the Cedar River indicate acetanilide degradates are transported into the alluvial aquifer in a manner similar to that indicated for atrazine and DEA. PMID- 10805244 TI - Changes in herbicide concentrations in Midwestern streams in relation to changes in use, 1989-1998. AB - Water samples were collected from Midwestern streams in 1994-1995 and 1998 as part of a study to help determine if changes in herbicide use resulted in changes in herbicide concentrations since a previous reconnaissance study in 1989-1990. Sites were sampled during the first significant runoff period after the application of pre-emergent herbicides in 1989-1990, 1994-1995, and 1998. Samples were analyzed for selected herbicides, two atrazine metabolites, three cyanazine metabolites, and one alachlor metabolite. In the Midwestern USA, alachlor use was much greater in 1989 than in 1995, whereas acetochlor was not used in 1989 but was commonly used in 1995. The use of atrazine, cyanazine, and metolachlor was approximately the same in 1989 and 1995. The median concentrations of atrazine, alachlor, cyanazine, and metolachlor were substantially higher in 1989-1990 than in 1994-1995 or 1998. The median acetochlor concentration was higher in 1998 than in 1994 or 1995. PMID- 10805245 TI - Comparative pathogenicity of different Actinobacillus suis O/K serotypes. AB - The pathogenicity of Actinobacillus suis serotypes O1/K1 (strain SO4), O1/K2 (strain C84), and O2/K2 (strain H91-0380) was evaluated in specific-pathogen-free (SPF) piglets challenged by intraperitoneal inoculation with approximately 1 x 10(7) colony-forming units per mL. All 3 strains produced peritonitis, but differences were observed in the composite histopathologic scores (P = 0.001) and in their ability to spread (P = 0.008) at 7 h post challenge. The O2/K2 strain caused the most severe peritonitis and disseminated most widely to other tissues. Moderate lesions were seen with the O1/K2 strain while the O1/K1 strain caused mild lesions and remained largely localized to the peritoneum. In an attempt to explain the basis of observed differences, the serum sensitivity of 9 A. suis strains with different O and K types was assessed. Regardless of the O/K type, all of the isolates tested were serum resistant. Moreover, most A. suis isolates grew as well or better in complement-replete sera as they did in complement depleted sera. These observations indicate that although 02 and K2 strains had a greater propensity to cause a disseminating septic inflammatory response in pigs, they were no more resistant to complement-mediated killing than O1 strains. PMID- 10805246 TI - A 24-kDa cloned zinc metalloprotease from Actinobacillus pleuropneumoniae is common to all serotypes and cleaves actin in vitro. AB - Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes proteases that degrade porcine hemoglobin and IgA in vitro. To further characterize A. pleuropneumoniae proteases, we constructed a genomic library expressed in Escherichia coli DH5alpha, and selected a clone that showed proteolytic activity. The recombinant plasmid carries an 800-base pair A. pleuropneumoniae gene sequence that.codes for a 24-kDa polypeptide. A 350-base pair PstI fragment from the sequence hybridized at high stringency with DNA from 12 serotypes of A. pleuropneumoniae, but not with DNA from Actinobacillus suis, Haemophilus parasuis, Pasteurella haemolytica, Pasteurella multocida A or D, or E. coli DH5alpha, thus showing specificity for A. pleuropneumoniae. The expressed polypeptide was recognized as an antigen by convalescent-phase pig sera. Furthermore, a polyclonal antiserum developed against the purified polypeptide recognized an A. pleuropneumoniae oligomeric protein in both crude-extract and cell-free culture media. This recombinant polypeptide cleaved azocoll, gelatin, and actin. Inhibition of the proteolytic activity by diethylpyrocarbonate suggests that this polypeptide is a zinc metalloprotease. PMID- 10805247 TI - Agar gel immunodiffusion test for the detection of bovine leukemia virus antibodies: lack of trans-Atlantic standardization. AB - Two agar gel immunodiffusion (AGID) kits for the serodiagnosis of bovine leukemia virus (BLV) were imported from Europe and were compared with North American kits. The BLV AGID kits from North America and from Europe differed significantly. The punches were different, as were the pattern distribution in the agar of the reference and the test sera, resulting in differences in the reading of the immunoprecipitation lines. Based on the testing of 1200 serum samples from cattle, the European kits gave a good correlation with the American kits, as indicated by their respective kappa values. However, the European kits were found to be less sensitive when evaluated against weakly positive samples from field specimens or following a dilution trial. Only 65% and 50% of the weakly positive samples detected by the American kit #1 were detected by the European kits #2 and #3, respectively. The American kit was also capable of detecting BLV antibodies in 45% of strongly positive samples diluted 1/50 in negative sera, while antibodies were detected in only 15% of the samples with the European kit #2 and in none of the samples with the European kit #3. False negatives were also detected with the European kits. Among the false negatives, the degree of expected reactions was weak (European kit #2) or of varying degrees of positivity (European kit #3). Besides the differences in format and performance, the BLV AGID kits in Europe are evaluated with the National Standard Serum E4 while a proficiency panel composed of a quadruplicate set of 10 reference sera is used in Canada to monitor the kits. Based on the overall observations, we noted a lack of standardization between the BLV-AGID kits used in North America and in Europe. PMID- 10805249 TI - Intestinal parasites of raccoons (Procyon lotor) from southwest British Columbia. AB - This is the first extensive survey of metazoan parasites (particularly of the roundworm Baylisascaris procyonis) from the intestines of raccoons in British Columbia. The sample collected in 1997-1998 consisted of 82 raccoons that had been sick or had been killed accidentally by automobiles. Fifteen parasite taxa were found: 3 nematodes, 9 digenetic trematodes, 2 acanthocephalans and 1 cestode. Ten of these parasites constitute new host records for raccoons, including 4 digenetic trematodes that have been reported in marine birds and mammals on the Pacific Coast of North America. Baylisascaris procyonis infected 61% of the raccoons with a mean intensity of 27. The high rate of infection indicates a large potential for environmental contamination and, thus, human and animal exposure to infectious eggs. Prevention of larva migrans is discussed, particularly for people in contact with raccoons in wildlife rehabilitation centers. PMID- 10805248 TI - Enzyme-linked immunosorbent assay for the diagnosis of bovine leukosis: comparison with the agar gel immunodiffusion test approved by the Canadian Food Inspection Agency. AB - Four commercially available bovine leukemia virus (BLV)-ELISA kits from Europe or the United States were compared to the agar gel immunodiffusion (AGID) test officially approved by the Canadian Food Inspection Agency (CFIA). A total of 1200 cattle serum samples were used. Three ELISA kits based on the envelope glycoprotein (gp51) gave an excellent correlation with the AGID test. The kappa values were 0.998, 0.984, and 0.986 for the ELISA kits #1, #2, and #3, respectively. The ELISA kit based on the p24 core protein was found to be less sensitive than the officially approved AGID test and detected 5.13% of false negatives. Forty BLV AGID strongly positive serum samples were diluted. Based on the dilution experiment, the gp51 ELISA kits were found to be more sensitive than the AGID test kits. They were capable of detecting antibodies in samples diluted up to 1/5000 (kit #1), 1/20 800 (kit #2) and 1/4000 (kit #3), whereas the AGID kit was only capable of detecting antibodies in samples diluted up to 1/100. Based on these observations, the gp51 BLV-ELISA was recognized as an official test method for the serodiagnosis of bovine leukosis in Canada. PMID- 10805250 TI - Direct MS-MS identification of isoxsuprine-glucuronide in post-administration equine urine. AB - Isoxsuprine is routinely recovered from enzymatically-hydrolyzed, post administration urine samples as parent isoxsuprine in equine forensic science. However, the specific identity of the material in horse urine from which isoxsuprine is recovered has never been established, although it has long been assumed to be a glucuronide conjugate (or conjugates) of isoxsuprine. Using ESI/MS/MS positive mode as an analytical tool, urine samples collected 4-8 h after isoxsuprine administration yielded a major peak at m/z 554 that was absent from control samples and resisted fragmentation to daughter ions. Titration of this material with increasing concentrations of sodium acetate yielded m/z peaks consistent with the presence of monosodium and disodium isoxsuprine-glucuronide complexes, suggesting that the starting material was a dipotassium-isoxsuprine glucuronide complex. Electrospray ionization mass spectrometry negative mode disclosed the presence of a m/z 476 peak that declined following enzymatic hydrolysis and resulted in the concomitant appearance of peaks at m/z 300 and 175. The resulting peaks were consistent with the presence of isoxsuprine (m/z 300) and a glucuronic acid residue (m/z 175). Examination of the daughter ion spectrum of this putative isoxsuprine-glucuronide m/z 476 peak showed overlap of many peaks with those of similar spectra of authentic morphine-3- and morphine-6 glucuronides, suggesting they were derived from glucuronic acid conjugation. These data suggest that isoxsuprine occurs in post-administration urine samples as an isoxsuprine-glucuronide conjugate and also, under some circumstances, as an isoxsuprine-glucuronide-dipotassium complex. PMID- 10805251 TI - Comparative effects of the human protein C activator, Protac, on the activated partial thromboplastin clotting times of plasmas, with special reference to the dog. AB - The commercial snake venom extract, Protac, is a specific activator of the anticoagulant zymogen, protein C (PC) in human plasma. This specific action has led to its use in developing coagulation-based and amidolytic-based assays for the diagnosis of quantitative and/or qualitative PC deficiency states in human beings. The purpose of the present study was to compare the effects of Protac on the activated partial thromboplastin times (APTT) of human, bovine, equine, and canine plasmas in order to determine the potential value of this venom extract as an activator in functional PC assays in these domestic animal species. As expected, Protac significantly prolonged the APTT of normal human plasma, but had no effect on plasma known to be devoid of PC. Clotting times were prolonged by 34%-214% with concentrations of venom activator ranging from 0.1-1.0 U/mL. Under identical conditions, Protac prolonged the APTT of equine plasma by 11%-98% over control times. Even more dramatic was the inhibitory effect of Protac on the clotting of bovine plasma, extending the APTT more than 3-fold at a venom concentration of 0.1 U/mL. At higher venom concentrations, most bovine plasmas remained unclotted after 300 s (control time 34.1 s). Under similar conditions, the canine APTT was unaffected by Protac, even when the venom concentration was increased to 3 U/mL. In order to determine the reason for the lack in response of canine plasma, the concentration of the APTT reagent was altered (decreased), exposure time of the plasma to the Protac was increased from 2 min to 9 min, and the plasma was diluted to assess for the potential existence of plasma PC inhibitors. Protac caused an unexpected shortening of the APTT when the contact activator reagent was diluted. Increasing the exposure time had no effect. Although a slight prolongation of the canine APTT was detected when the plasma was diluted, the presence of strong plasma PC inhibition was considered an unlikely cause of the lack of significant anticoagulant action. The failure of Protac to exert a strong inhibitory effect on the canine APTT, as well as to generate amidolytic activity, suggests that this venom extract does not stimulate the production of activated PC activity in canine plasma. This may result from molecular differences in the canine PC molecule that prevent the formation of the stoichiometric complex of venom extract, APTT reagent, and canine protein, a complex thought to be essential for the PC-activating function of Protac. Protac may be suitable as an activator of PC in bovine and equine plasmas; however, it appears ineffective in generating anticoagulant activity in canine plasma. PMID- 10805252 TI - Effect of clomipramine on monoamine metabolites in the cerebrospinal fluid of behaviorally normal dogs. AB - The tricyclic antidepressant, clomipramine, is an effective treatment for canine compulsive disorder (canine CD). This disorder is a clinical syndrome of abnormal conflict behaviors and its pathophysiology is unknown. However, because clomipramine is an effective treatment, information about the drug's neurochemical effect could enhance the understanding of canine CD. The following experiment used 6 behaviorally normal dogs to assess the effect of clomipramine (3 mg/kg, q24h, PO) on the central turnover of 3 monoamines (serotonin, dopamine, and norepinephrine) as measured by the concentrations of their respective metabolites in cerebrospinal fluid (CSF). In a randomized, placebo-controlled, AB BA crossover experiment, cisternal CSF was taken after 1, 2, 4, and 6 wk on each treatment. No effect of clomipramine was detected. This contrasts with human studies that have suggested that clomipramine affects the concentrations of monoamine metabolites in lumbar CSF. However, those papers do not address methodological assumptions, such as (i) metabolites in CSF originate only from the brain, and (ii) concentrations of metabolites in cisternal/lumbar CSF reflect the concentrations in local areas of the brain. Notwithstanding the small sample size, our results suggest that more localized sampling techniques (e.g. microdialysis) are needed when examining the effect of drugs on central monoamine metabolites. Clomipramine's efficacy for canine CD indicates the need for neurobiological research and, to our knowledge, our study is the first of its kind in dogs. The resulting data are preliminary but they can inform optimal neurobiological studies of canine CD. PMID- 10805253 TI - Type I interferon production in cattle infected with 2 strains of foot-and-mouth disease virus, as determined by in situ hybridization. AB - Four calves were exposed via aerosol to 1 of 2 strains of foot-and-mouth disease virus. Two animals received virus derived from an infectious clone virus (A12-IC) and 2 received virus derived from the same clone but which lacked the leader coding region (A12-LLV2) that codes for a protein responsible for turning off host protein synthesis. Animals were euthanized at 24 and 72 h post exposure. Cattle receiving A12-IC had a rapid course of disease with more virus in tissues while A12-LLV2-infected cattle did not develop clinical signs of disease. Formalin-fixed, paraffin-embedded tissue sections were probed with digoxigenin labeled riboprobes corresponding to the coding sequence for bovine interferon (IFN) alpha and IFNbeta. Staining for IFNalpha mRNA was noted in mononuclear cells of the lungs of all animals and in respiratory lymph nodes of cattle receiving A12-IC. Staining for IFNbeta mRNA was confined to bronchiolar epithelium and present only in the animals infected with A12-IC. Inability of the A12-LLV2 virus to achieve levels of spread seen with A12-IC may be related to translation of IFNalpha in A12-LLV2-infected cells, which renders adjacent cells less susceptible to productive infection. PMID- 10805254 TI - Susceptibility of piglets to rabbit hemorrhagic disease virus following experimental infection. AB - The possibility exists that rabbit hemorrhagic disease virus (RHDV) can be transmitted to swine, through lapinized hog cholera virus (HCV) vaccine. To investigate the infectivity of RHDV in swine, 16 four- to six-week-old piglets were inoculated subcutaneously with RHDV, and samples of liver, lung, spleen, kidney, bile, adrenal gland, tonsil, mesenteric lymph node, thymus, urine, buffy coat, and feces were collected from each of 2 animals on Days 0, 1, 2, 3, 5, 7, 14, and 28 post infection. Using reverse transcription-polymerase chain reaction, viral RNA was detected in most tissues by Day 3 and was absent after Day 5, except in lung and liver tissues, in which viral RNA was detected up to Day 14. Viral RNA was not detected in kidney, urine, feces or bile. Antibody responses, as detected by hemagglutination inhibition, were of low titer and short duration, and were similar in animals inoculated with viable RHD and in those given formalin-inactivated RHDV (n = 2). Neither viral RNA nor antibody were detected in the negative control or in the uninfected, in-contact animals. PMID- 10805255 TI - Protection studies on winter dysentery caused by bovine coronavirus in cattle using antigens prepared from infected cell lysates. AB - Cells infected with bovine coronavirus (BCV) were solubilized with Triton X-100 to yield a cell lysate (CL) antigen having high hemagglutinating (HA) titers. The antigen gave high HA titers using rat erythrocytes, suggesting that it contained large amounts of hemagglutinin esterase (HE) antigen. The CL antigen, combined with an oil adjuvant, was tested for protective and antibody-inducing activities in cattle. Four groups (2 cattle/group) of cattle were inoculated with CL antigen having HA titers of 16 000, 4000, 1000, and 250. Another group served as untreated controls. Two intramuscular inoculations were given at an interval of 3 wk. The animals were challenged with virus 1 wk after the second inoculation. The groups immunized with the CL antigen having an HA titer of 4000 or 16 000 produced hemagglutination inhibition (HI) antibody titers of > 320 and serum neutralizing (SN) antibody titers of > 1280. These groups of animals showed no clinical abnormalities after challenge. In the groups immunized with CL antigen at an HA titer of 1000 or 250, HI antibody titers were 40 to 160 and SN titers were 80 to 640. The cattle with HI antibody titers of > or = 160 and the SN titers of > or = 640 showed no clinical signs, but the cattle with the HI antibody titer < 80 and the SN antibody titer < 160 developed watery diarrhea and fever after challenge. These results indicate that CL antigen with high HA titer induces antibody production in cattle that provides effective protection against winter dysentery. PMID- 10805256 TI - Blood glycated hemoglobin evaluation in sick dogs. AB - Blood glycated hemoglobin concentration reflects long-term serum glucose levels in dogs. In this study, the effects of several diseases on blood glycated hemoglobin levels have been evaluated. For this study, blood samples were drawn from 93 unhealthy dogs. The animals were distributed into 10 groups according to pathological process (group 1, digestive problems; group 2, leishmaniasis; group 3, anemia; group 4, dermatological disorders; group 5, urinary problems; group 6, cardiorespiratory problems; group 7, diabetes mellitus; group 8, insulinoma; group 9, general diseases; group 10, control group). Blood glucose and glycated hemoglobin concentrations and hemoglobin and hematocrit values were analyzed in all the animals. In diabetic dogs, a strong increase in blood glycated hemoglobin was observed when compared with the other groups (P < 0.01). In contrast, dogs with insulinoma showed a decrease in blood glycated hemoglobin, though significant differences were not reported in all cases. No change in blood glycated hemoglobin concentrations were reported in dogs affected by other diseases. So, we can suppose that only the chronic alterations in glucose metabolism (chronic hyper- or hypoglycemia) can induce significant changes on the blood glycated hemoglobin concentrations in dogs. PMID- 10805257 TI - Allogeneic bone marrow transplantation for lymphoma. AB - Registry data show that use of allogeneic transplantation for non-Hodgkin's lymphoma, and to a lesser extent, Hodgkin's disease is increasing. Although no prospective randomized trials have been performed, most comparisons show a significantly lower relapse rate when allogeneic transplant results are compared to results of autologous hematopoietic stem cell transplantation. The lower relapse rate following allogeneic transplantation, as well as several other lines of evidence, support the existence of a graft-versus-lymphoma effect. Nevertheless, in most comparisons, the lower relapse rate following allogeneic transplantation is offset by higher transplant-related mortality. These results make it difficult to find situations where definite overall survival advantages associated with the use of allogeneic transplantation can be demonstrated. The use of low-intensity non-myeloablative regimens for allogeneic transplantation is attracting attention. It is hoped that this approach may harness a graft-versus lymphoma effect with less morbidity and mortality than conventional allogeneic transplantation, but more data are required to assess the value of this treatment. PMID- 10805258 TI - Monocyte-macrophage system as targets for immunomodulation by intravenous immunoglobulin. AB - Pooled human intravenous immunoglobulin (IVIg) has been used successfully to treat or ameliorate the clinical manifestations of humoral immune deficiencies, haematological disorders, HIV infection and many other diseases states. However, the mechanism of action of IVIg remains unclear. Several mechanisms of action of IVIg have been proposed. These include Fcy receptor blockade, accelerated clearance of endogenous pathogenic auto-antibodies, inhibition of components of the complement cascade, neutralization of super-antigens and bacterial toxins as well as anti-cytokine and anti-idiotype effects. A major contributor to host immunity and immune surveillance against infection, tissue or cell damage and malignancy is the monocyte/macrophage system. Monocyte-directed inflammation is a desirable consequence of microbiological or malignant challenge. However, monocyte hyperactivity may contribute to certain pathological conditions. These include the systemic inflammatory response syndrome (SIRS), septic shock, other dysregulated inflammatory disorders and auto-immunity. Novel therapies that can suppress the hyperactive state or correct monocyte/macrophage dysfunction without compromising normal host cell-mediated immunity are desirable. In this review, we discuss the immunomodulatory effects of IVIg focussing particularly upon the monocyte/macrophage system in pertinent disease states. PMID- 10805259 TI - Red cell antigens on band 3 and glycophorin A. AB - Band 3 and glycophorin A (GPA) are the two most abundant integral proteins of the red cell membrane, being present in approximately 10(6) copies per cell. The main functions of band 3 are membrane anion transport and maintenance of red cell membrane stability through interaction with the cytoskeleton. GPA plays an important role in prevention of red cell aggregation in the circulation and contribution to the glycocalyx. The extracellular domains of both proteins are highly polymorphic. Band 3 carries the antigens (currently 19) of the Diego blood group system and GPA and glycophorin B the antigens (currently 43) of the MNS system. There is substantial evidence that band 3 and GPA associate in the red cell membrane and the Wr(b) antigen, although a product of the band 3 gene, is known to require a complex of GPA and band 3 for normal expression. The discovery of a novel GPA mutation (Ala65-->Pro) giving rise to aberrant Wr(b) expression has been informative with regard to the site of interaction of the two proteins. The extensive array of GPA-related antigens is largely due to genetic events between two closely linked genes and different genetic mechanisms can give rise to the same antigen. This is in contrast to the antigens on band 3 which are exclusively due to single nucleotide mutations in the band 3 gene. PMID- 10805260 TI - RhD haemolytic disease of the fetus and the newborn. AB - When an RhD negative mother is exposed to the RhD positive red cells (usually as transplacental haemorrhage), she develops allo-anti-D which crosses the placenta and then results in the destruction of fetal red cells. Clinical manifestations of RhD haemolytic disease (HDN) range from asymptomatic mild anaemia to hydrops fetalis or stillbirth associated with severe anaemia and jaundice. HDN was a significant cause of fetal mortality and morbidity until the introduction of amniocentesis, intrauterine transfusion, controlled early delivery and exchange transfusion in the management of severely alloimmunised women and their fetuses. The objective of monitoring alloimmunised women is to identify fetal anaemia and prevent the development of life-threatening hydrops. Evaluation involves assessing the history of previous pregnancies; serial estimation of maternal anti D levels; serial ultrasound measurements; serial amniocentesis; fetal blood sampling, and intrauterine transfusion when indicated. Diagnostic genotyping by DNA-based methods can identify at-risk RhD positive fetuses early in gestation. Identification of transplacental haemorrhage (TPH) as the stimulus for anti-D antibody production led to the development of anti-D immunoglobulin prophylaxis for at-risk RhD negative women who are not already alloimmunised. Prevention includes administration of anti-D immunoglobulin for any event associated with TPH during pregnancy, and at delivery of an RhD positive infant. Prophylactic routine administration of anti-D immunoglobulin at 28 (and 34) weeks gestation, in addition to the above, has reduced alloimmunisation to <1% of RhD negative women carrying an RhD positive fetus. PMID- 10805261 TI - Allograft sling material: is it the state of the art? PMID- 10805262 TI - Constipation in Australian women: prevalence and associated factors. AB - A postal health survey was completed by 14761 young women (aged 18-23 years), 14070 middle-aged women (45-50 years) and 12893 older women (70-75 years). The prevalence of constipation was 14.1% (CI 13.5-14.7) in young women, 26.6% (CI 25.9-27.4) in middle-aged women, and 27% (CI 26.9-28.5) in the older women. The prevalence of hemorrhoids was 3.2% (CI 2.9-3.4 young), 17.7% (CI 17.1-18.4 middle aged) and 18.3% (CI 17.6-19.0 older). In the middle-aged and older women, those who reported previous gynecologic surgery were between 18% and 63% more likely to report constipation; in the younger cohort, women with one or two children were also more likely to report constipation (adjusted OR 1.43-1.46). One-third of the young women and half the middle-aged and older women had sought help for constipation; the majority indicated that they were satisfied with the help available to them. PMID- 10805263 TI - Relationship between the length of the perineum and position of the anus and vaginal delivery in primigravidae. AB - The aim of the study was to determine perineal length and anal position in primigravidae and to evaluate their effect on vaginal delivery. The distances between the fourchette and each of the center of the anal orifice and the inferior margin of the coccyx were measured in 212 primigravidae with singleton term pregnancies during the first stage of labor. Anal position index was calculated by dividing the first measurement by the second. The mean +/- SD length of perineum was 4.6 +/- 0.9 cm. The mean +/- SD anal position index was 0.49 +/- 0.12. Women with a short perineum (<4 cm) or a small anal position index (<0.42) had significantly higher rates of episiotomy, perineal tears and instrumented delivery. This association was also significant by multiple logistic regression analysis. It was concluded that a short perineum and anterior displacement of the anus were associated with traumatic vaginal delivery in primigravidae. PMID- 10805264 TI - Lower urinary tract symptoms in patients with Sjogren's syndrome and systemic lupus erythematosus. AB - Sjogren's syndrome (SS) and systemic lupus erythematosus (SLE) are autoimmune diseases which have many similarities with interstitial cystitis (IC), a urinary bladder disease with unknown etiology. This survey on the occurrence, severity and nature of lower urinary tract symptoms among patients suffering from SS or SLE showed that these patients have significantly more urinary complaints, especially irritative bladder symptoms, than age- and sex-matched controls. We studied 36 patients with SS, 85 patients with SLE and 121 controls. In these groups, 25%, 29% and 66%, respectively, were free of urinary symptoms. The prevalences of mild symptoms were 61%, 62% and 27%, and severe symptoms 14%, 9% and 7% in the respective groups. SS and SLE patients with urinary complaints reported mostly urinary frequency (27% and 62%) and suprapubic pain (36% and 34%). The most common symptom in the control group was stress urinary incontinence. The frequency of lower urinary tract problems in patients with SS and SLE supports the concept that autoimmune disorders also have bladder affections. PMID- 10805265 TI - MR imaging in posthysterectomy vaginal prolapse. AB - In the diagnostic work-up of vaginal prolapse after hysterectomy cystoceles can be identified by sonography, whereas enteroceles and rectoceles can only be suspected in a routine clinical setting. The present pilot study was undertaken to investigate the diagnostic role of magnetic resonance imaging (MRI) in the differentiation of cysto-, entero- and rectoceles in women with posthysterectomy vaginal prolapse. Thirteen women (mean age 61, SD +/- 7 years) with posthysterectomy vaginal prolapse underwent MRI (Gyroscan S 15, Philips). A median sagittal image series was obtained with a gradient-echo sequence, fast field echo, both at rest and during Valsalva maneuvers. MRI allowed the identification of cysto-, entero- and rectoceles, and differentiation between entero- and rectoceles in cases with inconclusive clinical findings. These findings make dissection more reliable and improve the outcome of hernia repair. No additional diagnostic information is obtained with MRI compared to ultrasound in the assessment of cystoceles. PMID- 10805266 TI - Abdominal urethrocystopexy using fibrin sealant. A prospective study of long-term efficacy. AB - Over the past decade efforts have been made to develop less invasive surgical treatment for female stress urinary incontinence (SUI). Abdominal urethrocystopexy with fibrin sealant combined with a couple of absorbable sutures has previously been reported as a promising method. This prospective observational study was aimed at evaluating the efficacy and safety of abdominal urethrocystopexy through a minilaparotomy using solely fibrin sealant (Tisseel) as the fixation glue. Forty-three women with objectively proven SUI were operated upon with this method. The subjective cure rates at 1 and 3 years' follow-up were 72% and 55%, respectively. The corresponding objective cure rates were 64% and 60%. No serious major operative complications occurred. One patient had transient urinary retention for 3 months. Otherwise, micturition was established within a median 1 day (range 1-3 days) after the operation. The result of this pilot study indicates a cure rate lower than that obtained with the conventional abdominal Burch colposuspension. Thus the method cannot be recommended as a standard procedure for treatment of SUI. PMID- 10805267 TI - Effect of intra-abdominal position of the bladder neck and stability of its supporting structures on pressure transmission ratio after colposuspension. AB - The aim of this prospective study was to evaluate the influence of the higher intra-abdominal position of the bladder neck and the stability of its supporting structures after colposuspension, on pressure transmission to the urethra at the level of the bladder neck. Twenty-eight patients were included in the study. The pressure transmission ratio (PTR) was calculated at the level of the bladder neck, whereas the position and mobility of the bladder neck during coughing were evaluated with perineal ultrasound examination. The measurements were performed before and 3 months after colposuspension. After colposuspension we found a significant elevation of the PTR (P=0.001), a significantly higher intra abdominal position (P=0.001) and decreased mobility (P=0.001) of the bladder neck during coughing. Also, a negative correlation between the elevation of PTR and decreased mobility of the bladder neck during coughing (r = -0.5049; P = 0.006) and a weak correlation between the elevation of PTR and a higher intra-abdominal position of the bladder neck during coughing were found (r = 0.3828; P=0.044). Reinforced tension resistance of the bladder neck supporting structures seems to be more important than intra-abdominal position of the bladder neck in achieving effective pressure transmission after colposuspension. PMID- 10805268 TI - Cystometry techniques in female infants and children. AB - Cystometry is increasingly used in children, being an important diagnostic tool for congenital and acquired malformations such as neurogenic bladder, vesicoureteric reflux, urinary tract infection, urge syndrome, nocturnal enuresis, urinary incontinence and anorectal malformations. During cystometry bladder storage and voiding function can be evaluated. Carefully conducted urodynamic studies provide an insight into the pathophysiologic mechanisms involved in voiding dysfunctions that cannot be obtained by any other diagnostic technique. A variety of methods are available, the most sophisticated being filling and voiding cystometry with flowmetry and electromyography (EMG) under fluoroscopy, i.e. videocystometry. A detailed home recording of the frequency and volume of micturition can provide important clues as to the underlying bladder dysfunction, and can significantly aid in the choice of appropriate investigative techniques, as well as in the interpretation of subsequent urodynamic findings. It must be realized that urodynamic studies are invasive procedures and that artifacts may occur and influence the correct interpretation of the results. Infants and children have a different spectrum of bladder dysfunctions from adults and are generally much less cooperative during a urodynamic study. Therefore, cystometric techniques must be significantly modified. This article reviews cystometry techniques and their application in female infants and children. Cystometry/flow/EMG studies with or without fluoroscopy, ambulatory urodynamics and telemetric urodynamic measurement and their application are outlined. PMID- 10805269 TI - Surgical treatment of interstitial cystitis in women. AB - Interstitial cystitis is a clinical entity that has been known for a century, but its pathophysiology remains largely unknown and the optimal treatment is a matter of ongoing discussion. A successful strategy for treatment relies on precise appraisal of symptoms, clinical findings and histology, as well as on the patient's individual personality. The least invasive treatment possible should be chosen, and only after conservative options have been exhausted should a surgical solution be considered. In this respect, anatomical bladder capacity plays an important role. A large capacity indicates the potential for conservative treatment and may be regarded as a negative predictor for the outcome of orthotopic bladder substitution. In contrast, a small anatomical capacity is unlikely to respond to conservative therapy, but is associated with a high probability of successful orthotopic bladder substitution. PMID- 10805270 TI - Effect of estrogen-progestogen hormonal replacement therapy on periurethral and bladder vessels. AB - This study assessed the effect of hormone replacement therapy using estrogens and/or progestogens on the number of vessels in the proximal and distal urethra, vesicourethral junction and bladder of castrated adult female rats. Forty-five virgin adult rats (Rattus norvegicus albinus) castrated for at least 30 days were used. They were assigned to five groups; group I (control) received no medication; the others received via the subcutaneous route, respectively, 17-beta estradiol (group II), medroxyprogesterone acetate (group III), a maize oil and benzyl acid solution - placebo (group IV) and 17-beta-estradiol combined with medroxyprogesterone acetate (group V), for a minimum of 28 days. Increased vascularization throughout the urinary tract, except in the distal urethra, was found following estrogen replacement alone. In the group that received combined estrogens and progestogens, no increase was found. It was concluded that estrogen replacement in castrated rats significantly increased the number of vessels in the lower urinary tract. PMID- 10805271 TI - The relationship of the in-situ advancing vaginal wall sling to vaginal epithelial inclusion cyst. AB - Epithelial inclusion cyst is an under recognized complication of the in-situ advancing vaginal wall sling. A 63-year-old woman with stage I pelvic organ prolapse and mixed incontinence underwent in-situ sling placement in November 1997. In February 1998 she presented with a painful recurrent inflammatory anterior vaginal wall mass. The mass was cystic and drained spontaneously four times over the period of conservative management. The patient underwent resection of a clinical and pathological vaginal epithelial inclusion cyst in September 1998. At 6-month follow-up the patient remains continent and the cyst has not reformed. The vaginal surgeon should be aware of the potential for epithelial inclusion cyst formation after in-situ sling placement, and actively search for them at postoperative examination. PMID- 10805272 TI - Endoscopic management of prolapsing intravesical ureterocele in an adult female- a case report. AB - The authors present a case of intravesical ureterocele in a female which prolapsed out of the external urethral meatus causing urinary obstruction, and was managed by reduction into the bladder followed by endoincision. PMID- 10805273 TI - A multicenter study of tension-free vaginal tape (TVT) for surgical treatment of stress urinary incontinence. PMID- 10805274 TI - Molecular and genetic mechanisms of factor XIII A subunit deficiency. AB - Factor XIII is a proenzyme for a plasma transglutaminase. Factor XIII in plasma is a tetramer (A2B2) held together by noncovalent bonds, and the A subunit contains the active site. Recently, the three-dimensional structure of the A subunit has been determined by x-ray crystallography. To understand the structure function relationships of the factor XIII molecule and its clinical implications in factor XIII deficiency, we characterized its genetic defects and closely examined its gene products, including mRNA and protein levels. A variety of missense and nonsense mutations (Arg260-Cys, Tyr283-Cys, Gly562-Arg) and deletions/insertions with or without out-of-frame shift/premature termination and splicing abnormalities (4-bp deletion with 464Stop, T insertion at the exon IV/intron D boundary with exon IV-skipping, 20-bp deletion at the exon I/intron A boundary) has been identified in cases demonstrating A subunit deficiency. In some cases, the A subunit mRNA levels were severely reduced. Their molecular and cellular bases have also been explored by expression experiments in mammalian cells and by molecular modeling. In most cases, impaired folding and/or conformational changes of the mutant A subunits lead to both intra- and extracellular instability, which is responsible for the A subunit deficiency in the patients. PMID- 10805275 TI - Studies on congenital protein C deficiency in Japanese: prevalence, genetic analysis, and relevance to the onset of arterial occlusive diseases. AB - Hereditary protein C deficiency is associated with a predisposition to venous thrombosis. We identified 43 patients with protein C deficiency by screening approximately 26,800 patients admitted to the National Cardiovascular Center Hospital. The observed prevalence of protein C deficiency was 1 per 620. We performed genetic analyses of 57 Japanese families with protein C deficiency. Combined with the results of the other studies in 10 families, the 67 Japanese families with protein C deficiency have been examined and 39 different gene defects have been identified. Some changes were solely identified in Japanese subjects, whereas others showed no such ethnic bias. The recurrent defects of Phe139Val, Arg169Trp, Val297Met, and Met364Ile substitutions and a G8857 deletion were found in 33 Japanese families, accounting for 49% of Japanese families with protein C deficiency, Finally, we examined the relevance of protein C deficiency to the onset of arterial occlusive diseases. In the examination of whether protein C deficiency hastens arterial occlusion, we found a significant difference (p = 0.02) in the age at onset of acute myocardial infarction between the patients with protein C deficiency (n = 10: 49.4 +/- 14.8 years) and a group of patients with normal protein C levels (n = 42: 60.5 +/- 10.6 years). At the onset of atherothrombotic cerebral infarction, the patients with protein C deficiency were significantly (p = 0.022) younger (n = 11:57.4 +/- 12.8 years) than those with normal protein C levels (n = 48: 64.6 +/- 10.1 years). Thus, we conclude that congenital protein C deficiency hastens the onset of arterial occlusive diseases, especially acute myocardial infarction, in Japanese subjects. PMID- 10805276 TI - Diagnosis of disseminated intravascular coagulation by hemostatic molecular markers. AB - In the present study, the positive rate of thrombin-antithrombin complex (TAT), plasmin-plasmin inhibitor complex (PPIC), soluble fibrin monomer (sFM), and D dimer for the diagnosis of disseminated intravascular coagulation (DIC) was evaluated. The study comprised 307 patients with DIC, 123 with pre-DIC, and 121 with non-DIC. Plasma levels of TAT, PPIC, sFM, and D-dimer were significantly higher in DIC and pre-DIC patients than in non-DIC patients. In DIC patients, the positive rate of sFM was high and that of D-dimer was low; the positive rate of PPIC was higher in patients with hematopoietic malignancy than in those without this disease. In pre-DIC patients, the positive rate of all markers was low (<0.16), and the positive rate of PPIC was relatively high. In non-DIC patients, the positive rate of all hemostatic markers was low (<0.16), that of sFM being the lowest. Scoring the positive rate of TAT, PPIC, and sFM disclosed the following results: 72% of DIC patients had three or more points, 17.6% of pre-DIC patients had three or more points, and almost all (96.6%) non-DIC patients had two or less points. Scoring the positive rate of TAT, PPIC, and D-dimer disclosed the following results: 52.9% of DIC patients and 27.4% of pre-DIC patients had three or more points and almost all (96.7%) non-DIC patients had 2 or less points. These data suggest that the combination of TAT, PPIC, and sFM is useful for making the diagnosis of DIC. PMID- 10805277 TI - Study of cardiovascular risk factors and hemostatic molecular markers in elderly persons. AB - In this study, plasma concentrations of hemostatic molecular markers were investigated in 118 elderly persons with normal renal function (aged 65 to 97 years) who could manage their activities of daily living (ADL) by themselves, to find a strategy for conservation or elevation of ADL and quality of life (QOL). In all subjects, the frequency by which hemostatic markers exceeded their upper limit of normal range was 35.9% for thrombin-antithrombin HI complex (> or = 3.7 ng/ml), 38.3% for soluble fibrin monomer (> or = 4.0 microg/ml), 41.8% for D dimer (> or = 1.0 microg/ml), 49.0% for plasmin-alpha2 plasmin inhibitor complex (> or = 1.0 microg/ml), and 53.7% for thrombomodulin (> or = 20 ng/ml). The mean plasma levels of these markers were slightly higher than the upper limit of their normal range. These markers were also investigated in samples of patients with and without cardiovascular risk factors and with and without cardiovascular diseases (ischemic heart disease and/or cerebral infarction). Furthermore, the results were analyzed in relationships between cardiovascular disease and cardiovascular risk factor or aging. The findings suggest that aging exerts a stronger influence on plasma levels of these hemostatic molecular markers than the presence of cardiovascular risk factors and cardiovascular diseases. From the viewpoint for conservation or elevation of ADL and QOL in elderly persons, also other factor, such as drug intake, lifestyle, aging, and so on, must be considered to clarify the relationship between the plasma levels of the hemostatic molecular markers and cardiovascular risk factors or cardiovascular diseases. PMID- 10805278 TI - Current status of hemophilia patients and recombinant coagulation factor concentrates in Japan. AB - The first recombinant factor VIII concentrate was introduced in 1987 to treat hemophilia A patients, and the product was licensed in the United States in 1992. More than 10 years have passed since the recombinant products have been used for treatment of hemophilia A. The new therapeutic options seem to be safe and effective through the gathered experiences. Recently, recombinant factor VIIa concentrate has become available to treat hemophilia patients with inhibitor and factor VII deficiency patients in Europe and also recombinant factor IX for the treatment of hemophilia B has been licensed in the United States and Europe. The usage of recombinant coagulation factors has expanded the routine therapy for hemophilia in many countries. In Japan, the consumption of recombinant factor VIII is increasing year by year, because many patients have started to think that the recombinant technology seems to be safe. Unfortunately, though, the factor VIIa and factor IX products have not been licensed yet in Japan. This article discusses the current status of patients with hemophilia and recombinant coagulation factor products in Japan. PMID- 10805279 TI - Antithrombotic regulation in human endothelial cells by fibrinolytic factors. AB - Vascular endothelial cells (ECs) modulate the blood fibrinolytic system by secreting tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), and their inhibitor, type-1 plasminogen activator inhibitor (PAI-1). ECs also express t-PA receptors (t-PAR) and u-PA receptors (u-PAR) on their cell surfaces, assembling both enzymes to regulate the cellular fibrinolytic activity. In addition, ECs modulate these factors in response to several stimuli. Fibrin clots on ECs induce the up- and downregulation of t-PA and PAI-1 production, respectively, thus causing an effective lysis of the fibrin clot. Heat shock (43 degrees C) increases the expression of u-PA, t-PA, PAI-1, and u-PAR by which ECs become more fibrinolytic around the cells. Furthermore, because ECs possess t-PAR and u-PAR on their cell surfaces, the binding of t-PA and u-PA is a critical event, which affords ECs the localized and condensed fibrinolytic potential. Therefore, ECs play a central role in antithrombotic activity by regulating the levels of these fibrinolytic factors. PMID- 10805280 TI - Coagulation-associated enhancement of fibrinolytic activity via a neutralization of PAI-1 activity. AB - Total fibrinolytic activity in the vasculature is finely tuned by the balance between tissue plasminogen activator and plasminogen activator inhibitor type 1 (PAI-1). Although PAI-1 targets plasminogen activators, it also reacts with other serine proteases such as thrombin and factor Xa. The latter was shown to interact with PAI-1 only when a physiological concentration of calcium ions (Ca++) is present. Through such interaction, thrombin and Ca++-bound factor Xa shortened fibrin clot lysis times in a purified system by neutralizing PAI-1 activity. Both unfractionated heparin and vitronectin were shown to enhance the clot lysis further. Together with the cleavage and inactivation of PAI-1 by human neutrophil elastase, which was reported previously from our laboratory, such neutralization of PAI-1 activity by these serine proteases was shown to be strongly involved in the coagulation-associated enhancement of fibrinolytic activity. PMID- 10805281 TI - Pathogenesis of a bleeding disorder characterized by platelet unresponsiveness to thromboxane A2. AB - A platelet disorder characterized by the absence of thromboxane A2 (TXA2)-induced platelet aggregation is a new clinical entity of platelet dysfunction. The platelets of three patients had the ability to bind exogenous TXA2, but synthetic TXA2 mimetic-induced postreceptor biochemical events, such as IP3 formation, Ca2+ mobilization, phosphatidic acid formation, and GTPase activities, were selectively defective, suggesting impaired coupling between the TXA2 receptor and phospholipase C activation. Gene analysis of the TXA2 receptor showed a substitution of Leu for Arg60 in the first cytoplasmic loop in all patients, and this mutation seemed to be responsible for this platelet disorder. PMID- 10805282 TI - Platelet activation mediated through membrane glycoproteins: involvement of tyrosine kinases. AB - Fc gamma RII cross-linking and anti-CD9 mAbs included tyrosine phosphorylation of Fc gamma RII, Syk, and Lyn associated with Fc gamma RII in Fc gamma RII cross linking but not in anti-CD9 mAb-induced platelet activation. We prepared various GST fusion proteins expressing one or two SH2 domains of Syk and evaluated the association between these GST fusion proteins with Fc gamma RII. Based on the results obtained from these experiments, we suggest that only one tyrosine residue in ITAM of Fc gamma RII is phosphorylated with anti-CD9 mAb and that both are phosphorylated with Fc gamma RII cross-linking. Platelet activation mediated by GPIb, the receptor for vWF, is also related with tyrosine phosphorylation. Botrocetin and vWF induced Syk activation. Shc was also rapidly and heavily tyrosine phosphorylated. Sre and Lyn, a 54-kDa tyrosine kinase, was associated with cytoskeletal proteins. When GPIb was immunoprecipitated with nonfunctional anti-GPIb mAbs after platelets were activated with vWF and botrocetin, an in vitro kinase assay revealed the transient association of a kinase activity with GPIb after platelet activation. Phosphoamino acid analysis of phosphorylated proteins in this assay demonstrated that only tyrosine residues but not serine or threonine were phosphorylated, suggesting that the kinase was indeed a tyrosine kinase. PMID- 10805283 TI - Molecular pathogenesis of Bernard-Soulier syndrome. AB - Bernard-Soulier syndrome (BSS), a hereditary qualitative platelet disorder, is now proved to be caused by a qualitative or quantitative abnormality of the platelet glycoprotein (GP) Ib/IX/V complex. We investigated the genetic background of two Japanese females with BSS and identified the molecular defects underlying this disease. The first case was a single base pair deletion within seven adenine repeats at position 4464-4470 (EMBL, no. M22403) of the GPIbalpha gene resulting in a frameshift and a premature stop codon. The second case was a single T-->C substitution at nucleotide 1856 (EMBL, no. M80478) of the GPIX gene resulting in Phe55(TTT)-->Ser(TCT) substitution. The latter case is of interest in considering the pathogenesis of BSS, because all four GPs consisting of the GPIb/IX/V complex have the same structural unit called leucine-rich repeat (LRR). Because this mutation is located within the LRR of the GPIX polypeptide, Phe55- >Ser substitution may result in an alteration of the LRR that leads to impaired surface expression of the GPIb/IX/V complex. To clarify the effect of this mutation on surface expression of the GPIb/IX complex, we performed transfection studies with a plasmid having this mutation. Mutant GPIX could not increase surface expression of the GPIb/IX complex, thus demonstrating an important role of the LRR of the GPIX polypeptide during biosynthesis. PMID- 10805284 TI - Angiogenesis and its regulation: roles of vascular endothelial cell growth factor. AB - Neovascularization is well known to occur in human atherosclerotic plaques, proliferative retinopathies, and malignant neoplasias. However, its pathophysiologic roles and mechanisms still remain unclear. In this study, histochemical examination of atherosclerotic plaques showed that vascular endothelial cell growth factor (VEGF) was expressed by the smooth muscle cells and foamy macrophages in the atherosclerotic intimas. The number of VEGF-positive cells was positively correlated with the number of intimal blood vessels. Moreover, VEGF gene transfer into rabbit carotid arteries using the hemagglutinating virus of Japan (HVJ)-liposomes showed that VEGF overexpression could induce the angiomatoid proliferation. In diabetic rats, VEGF was overexpressed in diabetic retinas, and thus the local overexpression of VEGF seemed to play an important role in the development of blood-retinal barrier breakdown in simple diabetic retinopathy. These results indicated that the VEGF can act as a local and endogenous regulator of endothelial cell functions and that VEGF induces the neovascularization under pathophysiologic conditions. On the other hand, the transfer of a decoy for the cis-element in the promoter region of the angiogenic factors would be an effective method for regulating angiogenesis, because other angiogenic factors' expression promoted by such cis element could be simultaneously suppressed. Therefore, this method may supply a useful therapeutic tool for the regulation of pathologic angiogenesis. PMID- 10805285 TI - Molecular biology of ryudocan, an endothelial heparan sulfate proteoglycan. AB - Ryudocan is a type I integral membrane heparan sulfate proteoglycan, which was originally cloned from rat microvascular endothelial cells. We have cloned the cDNA of rat ryudocan. The deduced amino acids of ryudocan has homologous transmembrane and intracellular domains with syndecan but very distinct extracellular regions. We also cloned the human ryudocan cDNA, of which the gene localizes on the chromosome 20q12. To better understand the regulation of ryudocan expression, we have determined the structural organization of the human ryudocan gene. The human ryudocan gene extends approximately 24 kb and is divided into five exons that appear conserved in syndecan family members. The 5'-flanking sequences of the human ryudocan gene contain a variety of potential binding sites for transcription factors and are capable of functioning as a promoter. We purified human ryudocan and evaluated its interactions with several extracellular ligands. It was found that basic fibroblast growth factor (bFGF), midkine, and tissue factor pathway inhibitor exhibited significant ryudocan bindings. Heparitinase, but not chondroitin ABC lyase treatment, destroyed those ryudocan bindings; thus, the heparan sulfate chains of ryudocan appear to be responsible for those bindings. Immunohistochemical analysis revealed that ryudocan is expressed in peripheral nerve tissues, fibrous connective tissues, and placental trophoblasts. These findings suggest that ryudocan may possess multiple biologic functions, such as bFGF modulation, neurite growth promotion, and anticoagulation, via heparan sulfate-binding effectors in the cellular microenvironment. PMID- 10805286 TI - cis-elements required for expression of human protein C inhibitor gene in HepG2 cells and its androgen-dependent expression in rat reproductive organs. AB - Protein C inhibitor (PCI) is a plasma serine protease inhibitor of activated protein C, which is the main protease of the anticoagulant protein C pathway. Human PCI is synthesized in the liver, kidney, and several reproductive organs (testis, seminal vesicle, and prostate). In the present study, we characterized cis-elements of the human PCI gene required for expression in the hepatoma derived cell line, HepG2 cells, and also evaluated rat PCI mRNA expression, particularly on the effect of androgen in rat reproductive tissues. On the PCI gene expression in HepG2 cells, transient expression assays using several deletion mutants and site-directed mutants of the human PCI gene and gel mobility shift assays using several synthetic oligonucleotides showed that the Spl-binding site (residues -302 to -294) and the upstream AP2-binding site (residues -350 to 343) play roles as the promoter and the enhancer, respectively. Both the A activator-binding site (residues -422 to -414) and the interferon-gamma response element (residues -164 to -157) serve as the silencer. In the study on PCI mRNA expression in the reproductive organs, we first cloned rat PCI cDNA and then evaluated the effect of androgen on the PCI mRNA expression. The isolated rat PCI cDNA contained a 1,218-bp coding region of a 406-amino acid precursor protein. The deduced amino acid sequence of rat PCI showed an 85.7% and 62.2% homology with that of mouse and human PCIs, respectively. Northern blot analysis showed that rat PCI mRNA was strongly expressed in the seminal vesicles, moderately in the testes, but not in the liver. PCI mRNA expression in seminal vesicles and testes increased during the process of development. The PCI mRNA expression in seminal vesicles was significantly decreased after castration or after 17beta estradiol treatment. Treatment with testosterone in the castrated rats significantly enhanced its mRNA expression. These findings suggest that the PCI gene expression in rat seminal vesicles is under androgen control. PMID- 10805287 TI - Effects of beta2-glycoprotein I and monoclonal anticardiolipin antibodies on extrinsic fibrinolysis. AB - Antiphospholipid antibodies (aPLs) are associated with an increased incidence of thrombosis, but the mechanisms responsible for thrombosis are unclear. The present study investigated the effect of both beta2-glycoprotein I (beta2-GPI) and aPLs on the activity of extrinsic fibrinolysis. The remaining tissue plasminogen activator (t-PA) of the sample consisting of beta2-GPI, two-chain recombinant t-PA, plasminogen activator inhibitor (PAI) -1 was measured by a chromogenic assay using synthetic substrate S-2251, Glu-plasminogen, and soluble fibrin monomer. Without PAI-1, beta2-GPI did not affect t-PA activity. When 14.3 ng/ml PAI-1 was added to 3.6 U/ml t-PA, the remaining t-PA activity was increased from 48.9% to 60.4% by the addition of beta2-GPI (190 microg/ml). The effect of beta2-GPI did not require phospholipids. The beta2-GPI seems to protect t-PA activity from the inhibition by PAI-1. When monoclonal anticardiolipin antibodies (aCLs), EY1C8, and EY2C9, which were established from a patient with antiphospholipid syndrome, were further added to the mixture with a diluted phospholipid (Platelin) to investigate the influence of aPL, the remaining t-PA activity decreased to 50.1 and 80.7%. Monoclonal aCLs appeared to inhibit the effect of beta2-GPI, that is, these monoclonals inhibited the fibrinolytic activity by an elevation in PAI-1 activity. These results suggest the possibility that the impairment of fibrinolytic activity by aCLs is one of reasons for the increased incidence in thrombosis in patients with aCLs. PMID- 10805288 TI - Analysis of intron 13 microsatellite repeat polymorphism of the factor VIII gene by capillary electrophoresis. AB - To establish a rapid and automatic gene analysis method, we used capillary electrophoresis (CE) for the analysis of the intron 13 microsatellite repeat polymorphism (MRP) of the coagulation factor VIII gene for the diagnosis of hemophilia A. In the analysis of a 20-bp DNA ladder marker, the reproducibility evaluated using the relative standard deviation (RSD) of the relative migration time for one fragment of each fragment were less than 0.01%, whereas the RSDs of the actual migration time of each fragment were 0.1-0.3%. Thus, the appropriate internal standard should be mixed with the sample when CE resolves polymerase chain reaction (PCR) products. We next analyzed the intron 13 MRP evaluated with (CA)n repeats of the factor VIII gene using a 200-bp DNA fragment as the internal standard. The results showed that the PCR products from the intron 13 MRP could be resolved using CE, even with the repeat numbers of 20 and 21, which differ by only 2 bp. These results suggest that CE is a suitable method for analyzing PCR products for gene diagnosis. PMID- 10805289 TI - Whole blood prothrombin time using diluted tissue factor is shortened in spontaneous hypertensive rats. AB - Clinical screening tests for blood coagulation that use plasma as samples cannot estimate the participation of platelets, leukocytes, and erythrocytes in blood coagulation system. We developed an assay to evaluate the total coagulation ability of blood and whole blood prothrombin time (WPT) using the principle of prothrombin time with the diluted-tissue factor as a trigger and a newly developed apparatus, STA, viscosity change detection system (electromagnetic clot detection). The activation of platelets by Ca ionophore shortened WPT and increased the expression of CD62P on the platelet surface. WPTs in citrated blood of spontaneous hypertensive rats (SHR) were significantly shorter than those of controls, Wistar Kyoto rats (WKY). Plasma levels of thrombin-antithrombin III (TAT) in SHR were significantly higher than those of WKY. Moreover, WPT and TAT levels were significantly correlated. Based on these results, WPT was found to be useful to estimate the activation of blood coagulation in whole blood. PMID- 10805290 TI - Fibrin D-dimer in thrombogenic disorders. AB - Measurement of D-dimer (fibrin degradation product) is important for determining not only the activation of fibrinolysis but also the severity of a hypercoagulable state. However, fibrin degradation products are in variable, and the reactivity to cross-linked fibrin degradation products produced during fibrin degradation differs depending on the kind of antibody used against D-dimer. In patients with disseminated intravascular coagulation or earthquake-induced mental and physical stress and in patients after percutaneous transluminal coronary angioplasty, all of which are associated with acute fibrin formation and degradation, some discrepancies between two methods of D-dimer detection, automated latex agglutination assay (LPIA) and enzyme-linked immunosorbent assay (Stago), were found. No discrepancies in persistent fibrin formation and degradation were found among the healthy elderly, patients with lacunar stroke, and patients with coronary artery disease, almost all of whom had levels under 5.0 microg/mL, as determined by both methods. Evidence of persistently increased intravascular coagulation and fibrin turnover in patients with atherosclerotic disease was found. The cleavage of cross-linked fibrin by plasmin results in a production of fibrin degradation products, mostly contained D-dimer domains. Although the clinical utility of D-dimer can be achieved by their detection with specific antibodies, measurement of D-dimer as high-molecular-weight fragments may be useful to determine whether patients will undergo further fibrin degradation. When intermediate products of the degradation process need to be assessed, D-dimer level measurement by LPIA may serve as a suitable marker for ongoing fibrinolysis. PMID- 10805291 TI - ISI/INR system in Japan: experience from simultaneous measurement of the same plasma at four different laboratories. AB - In 1984, the Scientific and Standardization Committee (formerly ICTH) recommended the use of the International Sensitivity Index and International Normalized Ratio (ISI/INR) System for the monitoring of oral anticoagulant therapy. This system was introduced because the sensitivity of thromboplastin reagents used for the measurement of prothrombin time (PT) was widely different and comparison among hospitals employing different reagents was virtually impossible. In this study, we simultaneously measured the plasma from 7 patients with warfarin therapy at 4 different institutions for PT seconds, PT-INR, thrombotest (TT) seconds and TT INR. The comparison between these laboratories revealed clinically important variances between the 4 laboratories even when PT was converted to PT-INR. Laboratory 1 and laboratory 3 were using the same thromboplastin reagents for the measurement of PT. The PT (seconds) in both laboratories showed similar numbers, but when they converted into INR, the variances were significant (maximum coefficient of variance 10.44). We investigated the reason why these differences occurred and found that the PT seconds (11.40) for normal control at laboratory 3 were somewhat larger than those of other laboratories. If we assume that PT-INR is identical to TT-INR, the estimated PT (second) for normal control at laboratory 3 can be calculated from TT-INR, and was found to be 10.56 +/- 0.10 seconds. This was nearly the same as the one that was used at laboratory 1. In conclusion, there still exist some difficulties that must be overcome before the ISI/INR system can be used reliably, and we suggest attention be given to the PT seconds used as normal control plasma. PMID- 10805292 TI - Increased hemostatic molecular markers in patients undergoing anticoagulant therapy. AB - We evaluated several molecular markers of hemostasis in 92 patients with hypercoagulable states treated with anticoagulant therapy. In all patients, the average values of the international normalized ratio (INR) were 1.70 +/- 0.50; this increase in INR was not, however, significant in patients under thrombotest (TT) monitoring. There were no thrombotic or severe bleeding complications in these patients during a period of 27 months. Plasma levels of thrombin antithrombin complex (TAT), plasmin-plasmin inhibitor complex (PPIC), D-dimer, and soluble fibrin monomer (sFM) were slightly increased, suggesting that anticoagulant therapy was not completely effective in our Japanese patients based on the values of the TT. The INR was negatively correlated with TT, protein C, and protein S and particularly with TT between 10 and 80%. The range of TT was not correlated with the plasma level of TAT, PPIC, D-dimer, or sFM, but the range of INR was correlated with the plasma level of TAT, D-dimer, and sFM. The percentage of TAT, D-dimer, and sFM within normal range was significantly lower in patients with high INR. These findings show that INR is better than TT for the monitoring of warfarin therapy and that the therapeutic values of INR during the anticoagulant therapy should be > 1.7 in Japanese patients. PMID- 10805293 TI - Differences in reactivity to vitamin K administration of the vitamin K-dependent procoagulant factors, protein C and S, and osteocalcin. AB - Vitamin K is a trace nutrient necessary not only for the synthesis of four plasma clotting factors but also the production of two important anticlotting factors, protein C and protein S, and the synthesis of two bone proteins. If protein C and protein S are produced more quickly and/or in higher quantities than four plasma coagulation factors after vitamin K administration, then the result is unfavorable for stopping of hemorrhage. We therefore studied the difference of time dependence of prothrombin procoagulant factors, protein C and S and bone Gla protein after the administration of vitamin K in normal and vitamin K-deficient neonates. Results of our study showed that, on the whole, coagulation factors increased markedly more than anticlotting factors after vitamin K administration. Furthermore, the increase in bone Gla protein was also higher compared with protein C activity, although the detailed mechanism of the difference in reactivity of prothrombin procoagulant factors, protein C and S and bone Gla protein to vitamin K administration is not clear. PMID- 10805294 TI - Five-year history of the Northeastern Society of Plastic Surgeons. PMID- 10805295 TI - The influence of air bags and restraining devices on extremity injuries in motor vehicle collisions. AB - The influence of air bags and other restraining devices on injury after motor vehicle collisions is not well defined. This study examined the relationship between the use of restraining devices and the incidence of extremity injuries in motor vehicle collisions. A retrospective analysis was performed on motor vehicle collision data submitted to the Pennsylvania Trauma Outcome Study database from 1990 through 1995. Criteria for submission included trauma patients who were admitted to the intensive care unit, who died during hospitalization, who were hospitalized for more than 72 hours, or who were transferred in or out of the receiving hospital. A total of 21,875 patients met these criteria. These patients were analyzed for the presence or absence of upper and lower extremity injuries and were compared based on their use of restraining devices. Restraining devices were categorized into four groups: air bag alone, air bag and seat belt, seat belt or carseat without air bag, and no restraining device. Statistical analysis was performed using the chi-squared test of association. For contingency tables with small expected frequencies, Fisher's exact test was used. Study participants included 11,688 men and 10,185 women with a mean age of 38 +/- 20 years. There were 16,033 drivers and 5,842 passengers. Air bags were deployed in 472 instances. In 297 of these cases, additional restraint was provided with a seat belt. In 6,632 cases, air bags were not deployed; however, patients were restrained with either a seat belt or a carseat. In 14,771 cases, patients were not restrained. When comparing restraining devices as a group vs. no restraint, there was a significant decrease in the incidence of upper (p = 0.018) and lower (p < 0.001) extremity injuries. Air bags, however, were associated with an increased incidence of both upper (p = 0.033) and lower (p = 0.002) extremity injuries when compared with no restraint or when compared among patients who were restrained. As a group, restraining devices decrease the incidence of upper and lower extremity trauma sustained by patients injured in motor vehicle collisions. Air bags, however, are associated with an increased incidence of upper and lower extremity injuries when compared with seat belts alone or when no restraining devices are used. PMID- 10805296 TI - The increase in TRAM flap survival after delay does not diminish long term. AB - The delay phenomenon is sometimes relied on to increase survival of the transverse rectus abdominis musculocutaneous (TRAM) flap in high-risk patients. TRAM reconstruction has then been performed traditionally 1 to 2 weeks after the delay procedure. The optimum time course from this surgical delay to flap elevation, as it relates to survival, has not been well studied--specifically whether the potential benefit of delay lessens at any particular time after the delay procedure. The authors examined the effects of varying time periods (up to 7 months) after delay, via division of the dominant bilateral superior epigastric vessels, on the viability of the TRAM flap in a rat model. TRAM flap survival improved significantly (p < 0.01) at all time points in delayed groups compared with a nondelayed control group. The benefit of delay in this model was maintained at all times, even long term. In conclusion, it may be possible to extend the delay period safely beyond the customary 1 to 2 weeks without compromising survival of the TRAM flap, which may prove to be more convenient. PMID- 10805297 TI - Endoscopic harvest of the rectus abdominis muscle. AB - The rectus abdominis muscle is a versatile muscle with many applications. The use of this muscle is often limited by its considerable donor site morbidity. This study reports a minimally invasive technique to harvest the rectus abdominis muscle. The described technique has been used successfully in 5 patients who required a superiorly based flap for reconstruction of a sternal defect. All patients have had long-term flap survival and resolution of their sternal osteomyelitis. Although initially lengthy, harvest times have been less than 1 hour for the last 3 patients. Patients report minimal discomfort at their operative site. To date there have been no hernias or other complications. The rectus abdominis muscle can be harvested successfully endoscopically. With no other modification other than port site placement, this technique could be used to harvest free flaps or harvest inferiorly based rectus flaps. This technique is learned easily, is safe, and should reduce substantially the donor site morbidity associated with more traditional harvesting techniques. PMID- 10805298 TI - Tumescent technique as an adjunct to breast implant removal and capsulectomy. AB - Hydrodissection has long been used in plastic surgery to facilitate dissection. The addition of epinephrine to the solution is thought to add to hemostasis, whereas the use of local anesthetic reduces intraoperative pain. Removal of breast implants and the associated capsules can be a tedious procedure with adhesions between the capsule and the parenchyma brought about by the prolonged presence of the device. The use of a mixture of Ringer's lactate and lidocaine with epinephrine has been used in this group of patients. Its use is described along with the benefits and pitfalls of the technique. PMID- 10805299 TI - Prophylaxis against Frey's syndrome in parotid surgery. AB - In 1990 the authors reported their preliminary study of the prevention of Frey's syndrome in 55 patients utilizing a superficial musculoaponeurotic system (SMAS) flap in parotid gland surgery. During the past 10 years, numerous studies have supported their original thesis that interposition of living tissue between the resected gland bed and the skin could prevent the development of this complication. The authors have expanded their own patient population and now consider this a definitive study on the prevention of Frey's syndrome. A total of 160 patients are presented with a follow-up period of 5 to 22 years. All patients underwent subtotal or total parotidectomy performed by one of the authors. A history was acquired and testing for Frey's syndrome (Minor's starch iodine test) was performed. As a result of this approach, and in spite of the intensive search for it, no cases of Frey's syndrome were encountered. The hoped-for secondary benefit of preventing the postparotidectomy retromandibular depression was somewhat less satisfactory, although most patients remain satisfied with their appearance. The debilitating symptoms in Frey's syndrome, which is reported to have an incidence of 5% to 50% in the typical parotidectomy patient, can be avoided with thoughtful preoperative planning. The authors favor an aesthetic incision followed by the development of an SMAS flap. The parotidectomy is then performed using the surgeon's preferred technique. The SMAS flap is then placed into the bed of the resected parotid gland. This institutes a protective tissue barrier guarding against the aberrant anastomotic communication between the postganglionic secretomotor fibers intended for the parotid gland, and the now adjacent sweat glands. Their patient population is large enough to provide significant evidence that Frey's syndrome can be prevented, compared with a meta analysis of parotid patients in multiple other studies in the literature. Assuming the patient's history and pathology does not preclude its use, the SMAS flap should be considered the standard of care for preventing Frey's syndrome in the postparotidectomy patient. If the SMAS flap is not available, a temporoparietal fascial flap has proved to be a good alternative. PMID- 10805300 TI - Oral sedation allows for comfort and amnesia during office surgery. AB - Surgical procedures commonly associated with parenteral sedation or general anesthesia were completed in an office operating room with local anesthesia and oral medication. Diazepam and alprazolam were used in combination to produce a relaxed state, allowing local anesthesia to be instilled. Surgery was completed successfully in all patients, and no untoward effects were seen in the patients. Amnesia was associated with the drug combination, and a protective action of the long-lived drugs against lidocaine toxicity is theorized. PMID- 10805301 TI - Microvascular anastomosis using 2-octyl cyanoacrylate in the rat femoral artery. AB - Patency of the microvascular anastomosis is the most important requirement for tissue survival in free tissue transfer and in replantation. In efforts to improve on the standard suture method of microvascular anastomosis, new techniques such as limited-suture sleeve anastomoses and histoacryl glue anastomoses have been employed experimentally. However, as a result of factors such as tissue toxicity and suboptimal outcome, cyanoacrylates have not enjoyed clinical use. In addition, sleeve anastomoses continue to utilize suture, increasing the risks of intimal damage, platelet adhesion, and thrombosis. In an attempt to surmount these problems, the authors investigated the use of a new 2 octyl cyanoacrylate glue and a sutureless sleeve anastomosis. Anastomosis of 20 rat femoral arteries with a sutureless sleeve technique bonded with glue resulted in an 80% patency rate at 1 day to 3 weeks. Failures occurred in the first few attempts as the technique was evolving. These encouraging results suggest that 2 octyl cyanoacrylate may have applicability in quick, sutureless microvascular anastomoses. PMID- 10805302 TI - Comparison of autologous fat transfer in fresh, refrigerated, and frozen specimens: an animal model. AB - The rising popularity of autologous fat transfer (AFT) has raised the questions whether cryopreserved adipocytes are a suitable substrate for soft-tissue augmentation, and which storage techniques may be most suitable for a maximally successful surgical result. An animal model of AFT was conceived using isogeneic Sprague-Dawley rats. Suction harvesting of fat and subcutaneous implantation was performed with specimens stored at -16 degrees C or 1 degrees C for a period of 1 or 2 weeks, as well as a group that underwent immediate implantation. A histological comparison of the experimental and control groups clearly demonstrated a decrease in viable adipocytes and an increase in signs of inflammation and fat cell necrosis in those animals that received stored fat instead of immediate fat implantation. These changes became more severe with increased length of storage and the use of refrigeration over freezing. The authors conclude that the practice of storing harvested adipocytes for later injection is not supported by the experimental evidence and should be avoided. PMID- 10805303 TI - "Pumping the regenerate": an evaluation of oscillating distraction osteogenesis in the rodent mandible. AB - Mandibular distraction osteogenesis (DO) has become an important technique to lengthen the hypoplastic mandible and to reconstruct osseous defects after ablative surgery. The hallmark of successful DO is the creation of new bone within the distraction gap. Several anecdotal reports have described alternating compressing and lengthening protocols (i.e., "pumping the regenerate") to augment regenerate bone formation. The purpose of this experiment was to analyze formally the effects of an alternating compression/distraction protocol with a traditional distraction protocol. Ten adult male rats underwent unilateral mandibular osteotomy with placement of a custom distractor. After a latency period of 5 days, distraction was initiated at a rate of 0.25 mm twice daily. Animals in the control group (N = 5) were distracted to a length of 5.0 mm for 10 days at a rate of 0.25 mm twice daily. In contrast, animals in the experimental group (N = 5) were distracted to a length of 2.5 mm (at a rate of 0.25 mm twice daily) for 5 days, then compressed 1.0 mm for a 2-day period, and redistracted to a length of 5.0 mm. Regenerate cross-sectional area was evaluated by computed tomography performed after 5 weeks of consolidation. Gross examination and histological analysis were performed by a panel of experienced reviewers. Radiological as well as histological analysis of regenerate cross-sectional area demonstrated no significant differences between experimental (i.e., "pumped") and control groups. Both groups demonstrated excellent regenerate bone formation with no evidence of fibrous union. This study represents the first attempt to investigate the anecdotal technique of pumping the mandibular regenerate. The authors have demonstrated that pumping the regenerate leads to no substantial differences in radiological or histological appearance of regenerate bone formation. PMID- 10805304 TI - Expression of adenovirally delivered gene products in healing osseous tissues. AB - Gene therapy has moved from the promise of laboratory investigation to the reality of clinical practice in just the last decade. Various methods for delivery of genes to host cells have been developed and utilized both in vitro and in vivo. From the perspective of the plastic surgeon, gene therapy holds the promise to augment healing in clinical situations that remain difficult to treat, such as chronic wounds, osteoradionecrosis, or possibly to expedite current clinical practices, such as distraction osteogenesis. The authors chose to investigate the potential for gene therapy in osseous tissues using a replication deficient adenovirus vector to deliver the marker transgene beta-galactosidase. An adenovirus vector is ideal for use in situations in which transgene expression is desired for only a relatively short period of time, such as wound and fracture healing. Utilizing a rat mandibular osteotomy model, they demonstrated that, using an adenoviral vector, foreign genes can be delivered in a simple fashion and can be expressed in a reliable manner within and around the osteotomy site for at least 10 days. Furthermore, there was no evidence of transfection of distant tissues associated with local application of the adenovirus vector. With this information, clinicians may now attempt to deliver osteogenic and angiogenic genes in a site-specific fashion to improve and expedite osseous healing. PMID- 10805305 TI - Hypoxia increases insulinlike growth factor gene expression in rat osteoblasts. AB - Vascular disruption secondary to fracture leads to a hypoxic zone of injury where the oxygen tension at the center of the wound is quite low. In this dynamic microenvironment, a number of growth factors are elaborated to stimulate the synthetic processes of fracture repair. Previously the authors have shown the hypoxia-induced increase of vascular endothelial growth factor expression in osteoblasts. The purpose of these experiments was to examine osteoblast expression of insulinlike growth factors (IGF) I and II--cytokines believed to play a role in increased collagen synthesis, chemotaxis, and proliferation of osteoblasts in response to hypoxia. Primary cell cultures of osteoblasts isolated from neonatal rat calvaria were subjected to hypoxia (PO2 = 35 mmHg) for 0, 3, 6, 24, and 48 hours. Northern blot analysis of ribonucleic acid (RNA) from resulting cultures demonstrated a more than 60% increase in IGF-II messenger RNA (mRNA) expression after 3 hours of hypoxia. IGF-II mRNA expression continued to increase through later time points to 200% and 260% of baseline at 24 and 48 hours respectively. In contrast, IGF-I demonstrated no significant change in mRNA expression compared with baseline control (normoxia) cultures. In these experiments the authors have demonstrated a hypoxia-induced increase in IGF-II but not IGF-I in primary osteoblasts. The differential expression of these two growth factors may underscore important differences in the behavior of osteoblasts in the hypoxic fracture microenvironment. Taken together, these data add additional support to the theory that hypoxia induces gene-specific changes in expression of molecules important to extracellular matrix formation for successful bone healing. PMID- 10805306 TI - Autologous cultured fibroblasts: a protein repair system. AB - Cultured autologous fibroblasts create a living injectable system that has been utilized effectively to treat rhytids, depressed scars, subcutaneous atrophy, acne irregularities, and laser wounds. Autologous cultured fibroblasts (Isolagen) as a protein repair system is produced from a 3-mm cutaneous punch biopsy. Skin biopsies are initiated in an in vitro tissue culture system. The cells are expanded to produce large quantities of fibroblasts and extracellular matrix totaling 1.0 to 1.5 ml. This cellular system is injected into the patient after 8 weeks of cellular expansion. Each area of treatment is injected once every 2 weeks for three injections. A total of 1,450 patients in the United States and Europe have been treated at designated centers. A total of 4,800 injections were given. At the University of Medicine and Dentistry of New Jersey and Hackensack University Medical Center, 94 patients were treated from 1995 through 1999. Long term follow-up ranged from 36 to 48 months. A subjective patient satisfaction survey showed 92% of the patients were satisfied with the grade of correction. A long-term follow-up survey revealed continuing improvement beyond the initial correction in 70% of patients. Results from other designated centers correlated with the authors' findings. Cultured autologous fibroblasts (Isolagen) appears to be a living, cellular, dynamic filler system capable of immediate correction and continued repair of dermal and superficial subcutaneous deficiencies. PMID- 10805307 TI - The use of newborn rats and an adenoviral gene delivery vector as a model system for wound-healing research. AB - An attractive experimental method to elucidate the role of growth factors and cytokines in cutaneous wound healing would be to overexpress or "knock out" a molecule using a gene delivery vector and observe the impact on the wound repair process. As a first step toward developing an adenoviral gene delivery procedure to study wound repair, the authors injected beta-galactosidase (beta-gal) adenoviruses either subcutaneously or intradermally into the dorsal skin of 10 day-old postnatal Sprague-Dawley rats. Histological analysis and beta-gal staining were used to determine the expression and localization of the transferred gene. Beta-gal expression was observed as early as day 1 and up to day 7 postintradermal injection and day 9 postsubcutaneous injection, with no obvious inflammatory reaction detected at the injection sites. Furthermore, as expected, greater beta-gal expression was observed in the dermis of intradermally injected rats compared with the dermis of subcutaneously injected rats. Next, the authors sought to determine whether cutaneous wounds would heal before dissipation of the transferred gene. They created incisional and excisional wounds on the backs of similar-age rats. They found that incisional wounds closed by day 5 postwounding, whereas excisional wounds closed by day 14 postwounding. Their study demonstrated that an adenoviral vector delivered a gene efficiently into newborn rat skin and maintained the gene expression for at least as long as it would take for an incisional wound to heal. The combined use of newborn rat wound models and an adenoviral vector may provide a useful in vivo system to define the biological roles of growth factors and cytokines involved in the wound repair process. These discoveries may lead to the development of gene therapy approaches for abnormal wound healing. PMID- 10805308 TI - Role of three-dimensional computed tomography in the assessment of nasoorbitoethmoidal fractures. AB - Successful management of nasoorbitoethmoidal (NOE) fractures requires early diagnosis and accurate determination of the extent of injury. Although the anatomic complexity of the NOE region merits a clear interpretation of the spatial relationships associated with each injury, the role of three-dimensional (3D) computed tomography (CT) in the assessment of these fractures remains unclear. To clarify the diagnostic value of 3D CT, this retrospective analysis compares statistically the accuracy of two-dimensional (2D) CT against 3D CT in the detection of NOE fracture line tracts in 21 consecutive patients with surgically confirmed NOE fractures. Accuracy of radiographic findings of both modalities were compared with intraoperative findings serving as the "gold standard" for detecting fracture lines in NOE fractures. This statistical analysis indicates that combining 3D CT and 2D CT produces a higher diagnostic yield in the evaluation of NOE fractures than using either modality alone. Although 2D CT was significantly superior (p < 0.05) for inspection of the medial orbital wall, 3D CT offered significantly greater diagnostic accuracy (p < 0.05) along the medial maxillary buttress, particularly at the piriform aperture. 3D CT clarified spatially several unusual NOE injuries that were indiscernible with 2D CT alone. We recommend the combination of 3D CT with 2D CT for serious NOE injuries, particularly those involving displacement of the NOE complex or associated with other major midfacial fractures. PMID- 10805309 TI - Extensor tendon rupture after internal fixation of a distal radius fracture using a dorsally placed AO/ASIF titanium pi plate. Arbeitsgemeinschaft fur Osteosynthesefragen/Association for the Study of Internal Fixation. AB - Extensor tenosynovitis has been reported as a complication of the Arbeitsgemeinschaft fur Osteosynthesefragen/Association for the Study of Internal Fixation (AO/ASIF) pi plate. The authors present a case of extensor tendon rupture after internal fixation with a pi plate after one of the screws in the distal limb loosened. This mechanism of extensor tendon rupture has not been reported previously. With appropriate prevention, monitoring, and intervention this complication could have been avoided. Schnur DP, Chang B. Extensor tendon rupture after internal fixation of a distal radius fracture using a dorsally placed AO/ASIF titanium pi plate. PMID- 10805310 TI - Correction of frontal bone hypertrophy with setback osteotomy and hydroxyapatite cement. AB - This case report illustrates a method of correction for severe hypertrophy of the frontal bone. Accurate preoperative assessment including physical examination, photographs, and radiological studies are necessary. The anterior table of the frontal bone is removed, recontoured, and stabilized to the adjacent bone while maintaining the integrity of the frontal sinus mucosa. Hydroxyapatite cement is used to resurface and contour the frontal bone. This method is demonstrated to be safe and well tolerated, and should be considered as the procedure of choice for this condition. PMID- 10805311 TI - Pure grisly acts. PMID- 10805312 TI - Re: Bourdonnement and other benign temporary breast implant sounds. PMID- 10805313 TI - Re: An unusual case of orbital hydatid cyst: a surgical emergency. PMID- 10805314 TI - Giant, aggressive basal cell carcinoma. PMID- 10805315 TI - Development of an immunoassay for the beta-adrenergic agonist ractopamine. AB - Antibody generated from ractopamine-hemiglutarate-KLH was used to develop a ractopamine ELISA. The antibody showed good sensitivity in phosphate buffer, with an IC50 of 4.2 ng/ml (ppb) toward ractopamine and 16.2 ng/ml toward glucuronides of ractopamine conjugated to the phenethanolamine phenol of ractopamine. Phenylbutylamine phenol glucuronides of the (RS, SR) ractopamine diastereoisomers showed about 4% cross-reactivity, but the glucuronide of the (RR, SS) diastereoisomers conjugated at the same phenolic group showed no detectable reactivity with the antibody. The antibody generally had cross-reactivity towards compounds with bis-phenylalkyl amine structures rather than compounds with simple branched N-alkyl substituents. For example, the antibody showed little or no cross reactivity towards clenbuterol, isoproterenol, metaproterenol, and salbutamol, but cross-reacted with dobutamine. The system demonstrated a matrix effect similar to other enzyme immunoassays, dilution of urine decreased but did not eliminate the matrix effect. PMID- 10805316 TI - Detection of interleukin-12 (IL-12)-secreting cells in normal mice with enzyme linked immunospot (ELISPOT) assay. AB - To assess mice interleukin-12 (IL-12)-secreting cells at a single cell level, we have developed a murine IL-12 specific enzyme-linked immunospot (ELISPOT) assay. The application of the newly developed method clearly showed the frequency of IL 12-secreting cells in the resident peritoneal exuded cells was higher than other organs of normal DBA/1J mouse. Moreover, we determined the frequency of IL-12 secreting cells in the spleens of five strains of inbred mice, and found the incidence of IL-12 secretors in the strain C57BL/6 to be greatest, and significantly greater than four of the others. These results are compatible with the predicted evidence, supporting this ELISPOT assay for IL-12-secreting cells is accurate. The procedure provides a useful tool for investigating complicated immune responses at a single cell level. PMID- 10805317 TI - One step enzyme linked immunosorbent assay for direct estimation of serum cortisol. AB - One step competitive enzyme linked immunosorbent assay (ELISA) for direct estimation of cortisol in human serum is described. Cortisol-3-O-carboxymethyl oxime-bovine serum albumin (cortisol-3-O-CMO-BSA) was used as an immunogen and cortisol-21-hemisuccinate-horse radish peroxidase (cortisol-21-HS-HRP) was used as a tracer. To the cortisol antibody coated microtiter wells, standards or serum samples (25 microl) along with cortisol-HRP conjugate (100 microl) were incubated for 2 hours at 37 degrees C. Bound enzyme activity was measured by, using TMB/H2O2 as a substrate. In this new strategy, chilled acetone stripped pooled human serum and sodium salicylate were used for preparing the standards and blocking the cortisol binding globulin (CBG), respectively. The sensitivity of the assay was .28 microg/100ml. The intraassay and interassay coefficient of variations (CVs) were ranged from 1.3% to 9.3% and 6.8% to 12.3 %, respectively. The analytical recoveries were 94% to 101.5%. The serum cortisol values, obtained by this method were correlated well with those, obtained by radioimmunoassay; r=0.95 (n=52). PMID- 10805318 TI - Investigation of electroplated conducting polymers as antibody receptors in immunosensors. AB - Antibodies raised against the conducting polymer, carbazole as a hapten, react to modulate the polymer's electrochemistry. Using cyclic voltammetry the reaction of the antiserum was discovered to influence the polymer matrix's electrochemistry by an amperometric response. It is suggested that these observation form the basis of a direct sensor for immunoassay. PMID- 10805319 TI - Direct micro-radioimmunoassay of the new renin inhibitor CGP 60536. AB - A solid phase method for direct radioimmunoassay in plasma of the new renin inhibitor CGP 60536 has been developed which does not require the extraction of the parent drug with organic solvents. The assay showed a good reproducibility down to plasma concentrations of 0.15 ng/ml (LOQ) with intra- and inter-assay coefficients of variation < or = 20%. The procedure, which requires only small volumes of plasma (25 microl), is simple to use and well suited for routine analysis. The method allows the investigation of the pharmacokinetics of CGP 60536 in animals and man given low oral doses of the drug. PMID- 10805320 TI - Generation of rhythmical ingestive activities of the trigeminal, facial, and hypoglossal motoneurons in in vitro CNS preparations isolated from rats and mice. AB - The central pattern generator (CPG) for masticatory movements has been located in the medial bulbar reticular formation, by using cortically induced rhythmical jaw and tongue movements as a model. To analyze how stationary input from the cortex is transformed into rhythmical output in the neuronal population comprising the CPG, rhythmical neural activities representing rhythmical food ingestive movements were experimentally induced in vitro. Bath-application of N-methyl-D aspartate (NMDA) induced rhythmical activities in the trigeminal (V), facial (VII) and hypoglossal (XII) nerves of in vitro brainstem-spinal cord preparations isolated from newborn rats and mice. This paper will review evidence for the notion that (1) the rhythmical XII nerve activity represents rhythmical sucking movements; (2) the population of neurons critically involved in the rhythm generation of the XII nerve is localized in the ventromedial medulla oblongata on both sides, and can induce rhythmical XII nerve activity on the same side independently of each other; (3) the rhythmical activities in the V, VII, and XII motoneurons are induced by separate CPGs, which are located segmentally at the respective level of the V motor, VII, and XII nuclei. In addition, rhythmical masticatory-like EMG activity of jaw muscles can be induced by repetitive stimulation of the pyramidal tract in the in vitro brainstem isolated from adult mice together with the oral-facial structures. We propose that the in vitro brainstem preparation is a useful tool for longitudinal analysis of postnatal development of the central pattern generation of food ingestive movements, including conversion from sucking to mastication. PMID- 10805321 TI - Effects of aging on oxytalan fibre in mouse periodontal ligament. AB - The reactions of periodontal tissues to the mechanical stimulation of teeth seem to be different in patients of different ages. Although the effects of aging on other structures of the periodontal tissues, i.e., collagen fibre, bone, and cementum, have been reported, there is very little information available concerning the effects of aging on periodontal oxytalan fibre, which is reportedly the only elastic element in the human periodontal ligament. The purpose of this research was to clarify the aging related changes in the distribution of oxytalan fibres in the periodontal ligament of mice. Histological sections of the periodontal ligament of the lower first molar of 10-week-old (control group) and 1-year-old (aged group) ICR mice were specifically stained for oxytalan fibres with aldehyde fuchsin after preoxidization, and observed by light microscopy and confocal laser scanning microscopy (CLSM). In the aged group, oxytalan fibres seemed to be more prevalent around blood vessels in comparison with the control group. Furthermore, CLSM clearly showed that oxytalan fibres in the aged group were relatively more tortuous and complex than those in the control group. This tortuosity and complexity might imply that the oxytalan fibres in aged mice have lost a considerable amount of their original elasticity. While further study of these changes is required, these results may help orthodontists to establish better treatment strategies and methods for adult orthodontic patients. PMID- 10805322 TI - P2Y2 receptor elevates intracellular calcium concentration in rabbit eye suprachoroid. AB - Effects of ATP on the intracellular free calcium ion concentration ([Ca2+]i) in the rabbit eye suprachoroid were investigated by means of fura-2 microfluorophotometry. ATP application (10 to 100 microM) elicited a dose dependent biphasic [Ca2+]i-increase: a fast phase typically peaking within 30 s and a following slow plateau phase, which lasted during the presence of ATP. The slow plateau phase was markedly diminished by removal of extracellular Ca2+, whereas the fast phase remained. An inhibitor of Ca2+ release from the sarcoplasmic reticulum (TMB-8), an endoplasmic Ca2+-ATPase inhibitor (thapsigargin) and a phospholipase C (PLC) inhibitor (U-73122) diminished the fast phase. A P2 receptor antagonist (Suramin) inhibited the ATP-induced [Ca2+]i response. The potency order of ATP and related substances in producing the [Ca2+]i-elevation was UTP approximately equals ATP>ATP-gamma-S>ITP>ADP. beta,gamma-MethyleneATP, 2-methylthioATP and UDP evoked no response. This order is consistent with the P2Y2 receptor characteristics. Cross-desensitization between ATP and UTP excludes the co-existence of the other types of receptors. In conclusion, the ATP-induced [Ca2+]i-elevation in the rabbit eye suprachoroid was elicited by the Ca2+ release from the PLC-dependent, thapsigargin-sensitive intracellular Ca2+ storage sites by activating P2Y2 nucleotide receptors. PMID- 10805323 TI - Study of mandibular movements in mandibulectomy patients--border movements and functional movements during mastication, deglutition and speech. AB - In mandibulectomy patients who have not undergone surgical reconstruction, the remaining mandibular segment is unstable and often deviated. Its movements have low reproducibility during mastication, deglutition and speech. The purpose of this study was to clarify three-dimensionally the differences in mandibular movements for each of these oral functions in mandibulectomy patients with and without mandibular continuity. Four mandibulectomy subjects (Group I) without mandibular continuity and three subjects (Group II) with mandibular continuity were selected. Their mandibular movements were recorded using a jaw movement tracking device with six degrees of freedom. Each movement was assessed graphically at the virtual incisor point and the rotational angles of the mandible in the frontal, sagittal and horizontal plane were analyzed. The findings were as follows; 1. In Group I, the border movements at the virtual incisor point exhibited an irregular and asymmetric envelope deviated to the resected side in the frontal plane, whereas Group II exhibited a smooth and symmetric envelope. 2. In Group I, the rotational angles in the frontal plane during border movements and mastication, in all planes during speech, and in the frontal and horizontal plane during deglutition were significantly larger than in Group II. A comparison among border and all functional movements in mandibulectomy patients revealed characteristic movements in the rotation of the mandible in the frontal plane. It is suggested that the rotational angle of the mandible is a useful parameter for assessment of mandibular movements in mandibulectomy patients. PMID- 10805324 TI - The hippocampal sclerosis whodunit: enter the genes. PMID- 10805325 TI - New magnetic resonance imaging methods for cerebrovascular disease: emerging clinical applications. AB - During the 1990s, novel magnetic resonance imaging (MRI) techniques have emerged that allow the noninvasive and rapid assessment of normal brain functioning and cerebral pathophysiology. Some of these techniques, including diffusion-weighted imaging and perfusion-weighted imaging, have already been used extensively in specialized centers for the evaluation of patients with cerebrovascular disease. Evidence is now rapidly accumulating that both diffusion- and perfusion-weighted imaging, particularly when used in combination with high-speed MR angiography, will lead to improvements in the clinical management of acute stroke patients. Other novel MR techniques, such as spectroscopic imaging, diffusion tensor imaging, and blood oxygenation level-dependent functional MRI, have not yet assumed a definitive role in the diagnostic evaluation of cerebrovascular disease. However, they are promising research tools that provide noninvasive data about infarct evolution as well as mechanisms of stroke recovery. In this article, we review the basic principles underlying these novel MRI techniques and outline their current and anticipated future impact on the diagnosis and management of patients with cerebrovascular disease. PMID- 10805326 TI - Interleukin (IL)1beta, IL-1alpha, and IL-1 receptor antagonist gene polymorphisms in patients with temporal lobe epilepsy. AB - Proinflammatory cytokines, including interleukin (IL)-1beta, are known to modulate effects of neurotoxic neurotransmitters discharged during excitation or inflammation in the central nervous system (CNS). They also regulate development of glial scars at sites of CNS injury. To elucidate a genetic predisposition of temporal lobe epilepsy with hippocampal sclerosis (TLE-HS+), we studied polymorphisms in the IL-1beta, IL-1alpha, and IL-1 receptor antagonist (IL-1RA) genes in 50 patients with TLE-HS+ and in 112 controls. Fifty-three patients who had TLE without HS were also examined (TLE-HS-) as disease controls. The distribution of the biallelic polymorphism in the promoter region at position 511 of the IL-1beta gene (IL-1B-511) was significantly different both between TLE HS+ patients and controls and between TLE-HS+ and TLE-HS- patients. The differences were due to overrepresentation of the homozygotes for IL-1B-511*2, which is suggested to be a high producer of IL-1beta, in TLE-HS+ patients compared with both controls and TLE-HS- patients. In contrast, there was no difference between TLE-HS- patients and controls. Our data suggest that, in the homozygotes for IL-IB-511*2, minor events in development such as febrile convulsions could set up a cascade leading to HS. PMID- 10805327 TI - Diagnosis of new variant Creutzfeldt-Jakob disease. AB - As of December 31, 1998, 35 deaths had been attributed to new variant Creutzfeldt Jakob disease (nvCJD) in the United Kingdom, of which 33 cases had been neuropathologically confirmed and 2 classified as probable nvCJD. Fifteen cases were male and 20 female. The median illness duration was 14 months (range, 8-38 months) and the median age at death was 29 years (range, 18-53 years). The dinical features were consistent with previous descriptions. In nearly all cases, there were early psychiatric symptoms after a median period of 6 months ataxia developed, followed by involuntary movements and cognitive impairment. Electroencephalograms did not show the "typical" appearances found in sporadic CJD, about half the cases tested had a positive 14-3-3 immunoassay, and over 70% of cases had bilateral pulvinar high signal on magnetic resonance brain scanning. Prion protein gene analysis showed that all cases were homozygous for methionine at codon 129. Diagnostic criteria for nvCJD have been formulated, which have a high sensitivity and specificity. PMID- 10805328 TI - Fumaric aciduria: clinical and imaging features. AB - Fumaric aciduria (fumaric acidemia, fumarase deficiency) is a rare inborn error of metabolism caused by deficient activity of fumarate hydratase, one of the constituent enzymes of the Krebs tricarboxylic acid cycle. We describe the clinical and imaging features of this disease arising from a consanguineous pedigree in 8 patients in the southwestern United States. Thirteen patients have been previously described in the medical literature. Our patients presented with an early infantile encephalopathy with profound developmental retardation and hypotonia, and most experienced seizures. Previously unreported characteristics described here include structural brain malformations, dysmorphic facial features, and neonatal polycythemia. Magnetic resonance imaging showed multiple abnormalities, including diffuse polymicrogyria, decreased cerebral white matter, large ventricles, and open opercula. Fumaric aciduria should be included in the differential diagnosis of inborn errors of metabolism that cause cerebral malformations and dysmorphic features. The possibility that inborn errors of energy metabolism may cause structural malformations deserves increased recognition. PMID- 10805329 TI - Differential features of patients with mutations in two COX assembly genes, SURF 1 and SCO2. AB - We screened 41 patients with undiagnosed encephalomyopathies and cytochrome c oxidase (COX) deficiency for mutations in two COX assembly genes, SURF-1 and SCO2; 6 patients had mutations in SURF-1 and 3 had mutations in SCO2. All of the mutations in SURF-1 were small-scale rearrangements (deletions/insertions); 3 patients were homozygotes and the other 3 were compound heterozygotes. All patients with SCO2 mutations were compound heterozygotes for nonsense or missense mutations. All of the patients with mutations in SURF-1 had Leigh syndrome, whereas the 3 patients with SCO2 mutations had a combination of encephalopathy and hypertrophic cardiomyopathy, and the neuropathology did not show the typical features of Leigh syndrome. In patients with SCO2 mutations, onset was earlier and the clinical course and progression to death more rapid than in patients with SURF-1 mutations. In addition, biochemical and morphological studies showed that the COX deficiency was more severe in patients with SCO2 mutations. Immunohistochemical studies suggested that SURF-1 mutations result in similarly reduced levels of mitochondrial-encoded and nuclear-encoded COX subunits, whereas SCO2 mutations affected mitochondrial-encoded subunits to a greater degree. We conclude that patients with mutations in SURF-1 and SCO2 genes have distinct phenotypes despite the common biochemical defect of COX activity. PMID- 10805330 TI - Functional motor unit failure precedes neuromuscular degeneration in canine motor neuron disease. AB - Hereditary canine spinal muscular atrophy (HCSMA) features rapidly progressive muscle weakness that affects muscles in an apparent proximal-to-distal gradient. In the medial gastrocnemius (MG) muscle of homozygous HCSMA animals, motor unit tetanic failure is apparent before the appearance of muscle weakness and appears to be presynaptic in origin. We determined whether structural changes in neuromuscular junctions or muscle fibers were apparent at times when tetanic failure is prevalent. We were surprised to observe that, at ages when motor unit tetanic failure is common, the structure of neuromuscular junctions and the appearance of muscle fibers in the MG muscle were indistinguishable from those of symptom-free animals. In contrast, in more proximal muscles, many neuromuscular junctions were disassembled, with some postsynaptic specializations only partially occupied by motor nerve terminals, and muscle fiber atrophy and degeneration were also apparent. These observations suggest that the motor unit tetanic failure observed in the MG muscle in homozygous animals is not due to synaptic degeneration or to pathological processes that affect muscle fibers directly. Together with previous physiological analyses, our results suggest that motor unit failure is due to failure of neuromuscular synaptic transmission that precedes nerve or muscle degeneration. PMID- 10805331 TI - The motor cortex shows adaptive functional changes to brain injury from multiple sclerosis. AB - Although multiple sclerosis (MS) is an inflammatory demyelinating disease, there can be substantial axonal injury and loss. We therefore hypothesized that adaptive cortical changes may contribute to limiting functional impairment, particularly in the early stages of the disease. To test our hypothesis, we used functional magnetic resonance imaging (MRI) to characterize the localization and volumes of activation in the motor cortex during simple flexion-extension finger movements. There were differences in the patterns of cortical activation with movement between the 12 MS patients and the 12 normal controls. All patients showed greater relative supplementary motor area activation than did the normal controls. The relative hemispheric lateralization of sensorimotor cortex (SMC) activation decreased in direct proportion to the total cerebral T2-weighted MRI hyperintense lesion load. This appeared to be due primarily to increases in ipsilateral SMC activation with increasing lesion load in white matter of the hemisphere contralateral to the limb moved. The center of activation in the contralateral SMC was shifted a mean of 8.8 mm posterior in patients relative to controls, providing additional evidence for cortical adaptive responses to injury. The magnitude of this posterior shift in the SMC activation increased with greater T2 lesion loads. These observations demonstrate that cortical recruitment for simple finger movements can change both quantitatively and qualitatively in the SMCs of MS patients, suggesting that cortical reorganization or "unmasking" of latent pathways can contribute to functional recovery. These adaptive changes are another factor potentially limiting the strength of the relationship between MRI measures of pathology and clinical measures of disability. PMID- 10805332 TI - An association between migraine and cutaneous allodynia. AB - Recent animal studies on the mechanism of migraine show that intracranial pain is accompanied by increased periorbital skin sensitivity. These findings suggest that the pathophysiology of migraine involves not only irritation of meningeal perivascular pain fibers but also a transient increase in the responsiveness (ie, sensitization) of central pain neurons that process information arising from intracranial structures and skin. The purpose of this study was to determine whether the increased skin sensitivity observed in animal also develops in humans during migraine attacks. Repeated measurements of mechanical and thermal pain thresholds of periorbital and forearm skin areas in the absence of, and during, migraine attacks enabled us to determine the occurrence of cutaneous allodynia during migraine. Cutaneous allodynia is pain resulting from a nonnoxious stimulus to normal skin. In 79% of the patients, migraine was associated with cutaneous allodynia as defined, and in 21% of the patients it was not. The cutaneous allodynia occurred either solely within the referred pain area on the ipsilateral head, or within and outside the ipsilateral head. Cutaneous allodynia in certain well-defined regions of the skin during migraine is an as yet unreported neurological finding that points to hyperexcitability of a specific central pain pathway that subserves intracranial sensation. PMID- 10805333 TI - Three novel mutations (G27E, insAAC, R179X) in the ORNT1 gene of Japanese patients with hyperornithinemia, hyperammonemia, and homocitrullinuria syndrome. AB - Hyperornithinemia, hyperammonemia and homocitrullinuria (HHH) syndrome presents with various neurological symptoms, including mental retardation, spastic paraparesis with pyramidal signs, cerebellar ataxia, and episodic disturbance of consciousness or coma caused by hyperammonemia. We report three novel mutations in the mitochondrial ornithine transporter gene (ORNT1) of Japanese patients with HHH syndrome: a nonsense mutation (R179X) associated with exon skipping and a frameshift, a missense mutation (G27E), and an insertion of AAC between codons 228 and 229, leading to an insertion of the amino acid Asn. The ORNT1 gene consists of at least six exons, and all exon-intron junction sequences conform to the GT/AG rule. All 3 patients were homozygous for their respective mutations. This study confirms that defects in the ORNT1 gene cause the HHH syndrome and that the genetic basis in Japanese patients is heterogeneous. PMID- 10805334 TI - Alpha-synuclein and Parkinson's disease: selective neurodegenerative effect of alpha-synuclein fragment on dopaminergic neurons in vitro and in vivo. AB - Missense mutations in the alpha-synuclein gene were associated with a familial Parkinson's disease, and alpha-synuclein is a major component of Lewy bodies, the intracellular inclusions that neuropathologically characterize Parkinson's disease. We investigated the neurotoxic activity of the nonamyloid component (NAC) of senile plaque, the fibrillogenic fragment (61-95) of alpha-synuclein, in vitro and in vivo. Rat primary mesencephalic neurons were exposed for 6 days to low concentrations of preaggregated NAC (0.5-10.0 microM). The number of dopaminergic neurons and dopamine content were both reduced with no effect on the general viability of the cells. At higher concentrations (25-100 microM), the neurotoxic effect of NAC was extended to all neurons. Preaggregated NAC was also toxic on a PC12 dopaminergic cell line differentiated with nerve growth factor. The intracellular localization of NAC has been identified by the exposure of neuronal cells to fluorescent peptide. In vivo application of aggregated NAC in the substantia nigra induced loss of dopaminergic neurons. Our data illustrate the selective neurotoxic effect of NAC for dopaminergic neurons and support the central role of alpha-synuclein in the pathogenesis of Parkinson's disease. PMID- 10805335 TI - Glutamate uptake is decreased in platelets from Alzheimer's disease patients. AB - Because excitotoxicity may be involved in neurodegeneration in Alzheimer's disease, we investigated possible modifications of platelet glutamate uptake in AD patients. High-affinity glutamate uptake was studied in platelets from 35 Alzheimer's disease patients, 10 multi-infarct dementia patients, and 35 age matched normal controls; it was decreased by 40% in platelets from Alzheimer's disease patients compared with controls and with multi-infarct dementia patients. Platelet glutamate uptake could be used as peripheral marker of glutamatergic involvement and as adjunctive diagnostic tool in Alzheimer's disease patients. PMID- 10805336 TI - Benzodiazepine receptor binding in Huntington's disease: [11C]flumazenil uptake measured using positron emission tomography. AB - We used positron emission tomography and [11C]flumazenil to analyze the benzodiazepine receptor binding in symptomatic and asymptomatic carriers of the Huntington's disease gene. We found an inverse relationship between [11C]flumazenil and [11C]raclopride binding in the putamen of symptomatic patients, and interpret this result as GABA receptor upregulation. PMID- 10805337 TI - Serum elastase activity is elevated in migraine. AB - Migraine has been associated with diseases considered to be related to extracellular matrix disorders--in particular, cervical artery dissection. In this population-based study, we found a highly significant association between migraine and the activity of serum elastase, a metalloendopeptidase degrading specific elastin-type amino acid sequences. Such enzymes are involved in matrix degradation. This association was seen in both sexes and was stronger for migraine with aura. These findings could help in the understanding of why patients with migraine are at higher risk of stroke. Further study is needed to establish whether extracellular matrix abnormalities play a broader role in the pathophysiology of migraine. PMID- 10805338 TI - Evidence for infection with Chlamydia pneumoniae in a subgroup of patients with multiple sclerosis. AB - In a pilot study, we identified Chlamydia pneumoniae in the cerebrospinal fluid by polymerase chain reaction in 5 of 10 patients with definite multiple sclerosis (MS). In a second series, 2 of 20 patients with definite MS and 3 of 17 patients with possible/probable MS or MS variants, but none of 56 patients with other neurological, diseases were polymerase chain reaction-positive. We confirm that C. pneumoniae can be found in the cerebrospinal fluid of MS patients, but our rate of positive results is lower than in a recent report. PMID- 10805339 TI - Levodopa-induced dyskinesias in Parkinson's disease: is sensitization reversible? AB - Levodopa-induced dyskinesias (LIDs) in patients with Parkinson's disease are considered to result from the severity of dopaminergic denervation in the striatum, which is an irrevocable phenomenon, and sensitization induced by long term intermittent administration of levodopa. Taking advantage of the 64% reduction of levodopa treatment allowed in 12 Parkinson's disease patients by continuous high-frequency stimulation of the subthalamic nucleus, we evaluated the severity of parkinsonian motor disability and LIDs during two levodopa challenges performed before the surgical implantation of the stimulation electrodes and after 8.8 months of continuous bilateral subthalamic nucleus stimulation that was interrupted 2 hours before the levodopa test. Motor disability during the "off" and "on" drug periods was unchanged. The severity of LIDs during the "on" period and dystonia during the "off" period decreased by 54% and 62%, respectively. The reduced severity of LIDs in the absence of subthalamic nucleus stimulation demonstrates that the sensitization phenomenon resulting from long-term intermittent levodopa administration is partially reversible. PMID- 10805340 TI - Increased serum transferrin receptor concentrations in Friedreich ataxia. AB - Mitochondrial iron accumulation is thought to underlie the pathophysiology of Friedreich ataxia and may occur at the expense of cytosolic iron. Decreases in cytosolic iron induce expression of the transferrin receptor, some of which is released into the serum. Here, we demonstrate that serum transferrin receptor concentrations are increased in patients with Friedreich ataxia, which supports the hypothesis that it is a disease of abnormal intracellular iron distribution. PMID- 10805341 TI - Motor benefit from levodopa in spastic quadriplegic cerebral palsy. AB - We report on a 16-year-old girl with spastic quadriplegic cerebral palsy associated with premature birth and typical periventricular leukomalacia, who had a dramatic improvement in motor function after treatment with carbidopa/levodopa. Kinematic and electromyographic analyses of reaching movements demonstrate that levodopa decreased muscle co-contraction, decreased unwanted movements, and improved her ability to maintain a steady arm posture. These findings suggest that levodopa be considered as an adjunct therapy for the treatment of spastic quadriplegic cerebral palsy. PMID- 10805342 TI - X-linked vacuolar myopathies: two separate loci and refined genetic mapping. AB - X-linked vacuolar myopathies can be divided into two forms: one that is associated with cardiomyopathy and mental retardation (XVCM-MR) and a second form, termed X-linked myopathy with excessive autophagy (XMEA), that spares cardiac muscle and has no central nervous system involvement. In this article, we demonstrate linkage between XMEA and markers on chromosome Xq28 and assign the XMEA gene locus to the most telomeric 10.5 cM of chromosome X. We also show that XVCM-MR is not allelic to XMEA. PMID- 10805343 TI - Influence of mutation type and X chromosome inactivation on Rett syndrome phenotypes. AB - We screened 71 sporadic and 7 familial Rett syndrome (RTT) patients for MECP2 mutations by direct sequencing and determined the pattern of X chromosome inactivation (XCI) in 39 RTT patients. We identified 23 different disease-causing MECP2 mutations in 54 of 71 (76%) sporadic patients and in 2 of 7 (29%) familial cases. We compared electrophysiological findings, cerebrospinal fluid neurochemistry, and 13 clinical characteristics between patients carrying missense mutations and those carrying truncating mutations. Thirty-one of 34 patients (91%) with classic RTT had random XCI. Nonrandom XCI was associated with milder phenotypes, including a mitigated classic RTT caused by a rare early truncating mutation. Patients with truncating mutations have a higher incidence of awake respiratory dysfunction and lower levels of cerebrospinal fluid homovanillic acid. Scoliosis is more common in patients with missense mutations. These data indicate that different MECP2 mutations have similar phenotypic consequences, and random XCI plays an important role in producing the full phenotypic spectrum of classic RTT. The association of early truncating mutations with nonrandom XCI, along with the fact that chimeric mice lacking methyl-CpG binding protein 2 (MeCP2) function die during embryogenesis, supports the notion that RTT is caused by partial loss of MeCP2 function. PMID- 10805344 TI - In vitro glatiramer acetate treatment of brain endothelium does not reduce adhesion phenomena. PMID- 10805345 TI - The role of inheritance in sporadic Parkinson's disease. PMID- 10805346 TI - Association of primary central nervous system lymphoma with long-term azathioprine therapy for myasthenia gravis? PMID- 10805347 TI - 14-3-3 cerebrospinal fluid protein and Creutzfeldt-Jakob disease. PMID- 10805348 TI - Circulating antiganglioside antibodies are not associated with the development of progressive disease or cerebral atrophy in patients with multiple sclerosis. PMID- 10805349 TI - Effect of glatiramer acetate (Copaxone) given orally in human patients: interleukin-10 production during a phase 1 trial. PMID- 10805350 TI - No association between the NOS3 codon 298 polymorphism and Alzheimer's disease in a sample from the United States. PMID- 10805351 TI - Mutations in the neuroserpin gene are rare in familial dementia. French Alzheimer's Disease and Fronto-Temporal Dementia Genetics Study Groups. PMID- 10805352 TI - Antineutrophil cytoplasmic antibodies (ANCA) and systemic vasculitis: update of assays, immunopathogenesis, controversies, and report of a novel de novo ANCA associated vasculitis after kidney transplantation. AB - OBJECTIVES: To characterize antineutrophil cytoplasmic antibodies (ANCA), their major autoantigens, disease associations, and pathophysiology in systemic vasculitides. To describe a patient with a novel de novo ANCA-associated vasculitis after kidney transplantation. METHODS: We reviewed and compiled the literature on ANCA-related topics and systemic vasculitis. Laboratory and clinical data from a cadaveric kidney transplant patient who developed necrotizing vasculitis involving glomerular capillaries, with crescent formation associated with P-ANCA and myeloperoxidase, were analyzed. RESULTS: Large-scale multi-center testing of patient and normal sera by the European ANCA Assay Standardization Project using immunofluorescence assays and enzyme immunoassays indicate the assays have good sensitivity and specificity, and diagnostic utility for ANCA-associated vasculitis. A few investigations covering basic and clinical research with ANCA remain controversial: whether endothelial cells do or do not express a 29-kd neutral serine protease termed proteinase-3 (PR-3), the target of ANCA in most individuals with Wegener's granulomatosis, and whether anti myeloperoxidase (MPO) ANCAs recognize a restricted number of epitopes on MPO. This issue has relevance for using monoclonal antibodies to treat patients with vasculitis who have adverse effects from immunosuppressive drugs. The two allelic forms of FcgammaRIIa (H131/R131) and the two of FcgammaRIIlb (NA1/NA2) are discussed as possible inheritable genetic elements for vasculitic disorders and for signaling responses. Stimulatory and costimulatory molecules, and cytokine profiles of T lymphocytes are characterized to show that these cells are actively involved in the ANCA-associated vasculitides. The patient described had a de novo ANCA associated small vessel vasculitis which developed after renal transplantation. CONCLUSIONS: There have been significant advances in the development of sensitive and specific ANCA assays. The immunopathogenetic mechanism of ANCA involves the constitutive FcgammaRs, ligands, and signaling responses to activate cytokine-primed neutrophils. This may lead to the generation of reactive oxygen intermediates, degranulation, and secretion of intracellular granule contents, and ultimately inflammation and vasculitis. PMID- 10805353 TI - Behcet's disease in Familial Mediterranean fever: characterization of the association between the two diseases. AB - OBJECTIVES: Familial Mediterranean fever (FMF) is a genetic disease, characterized by attacks of fever and painful manifestations. Several vasculitides are more common in FMF than in the general population. The aim of the study was to define and characterize the association between FMF and Behcet's disease (BD), a form of vasculitis not previously related to FMF. METHODS: We conducted a retrospective study in which FMF patients, also suffering from BD (FMF-BD), were recruited from about 4,000 patients registered in our clinic, using a computer survey. Patients identified by the screening process were examined, and those meeting the published criteria for the diagnoses of FMF and BD were classified as FMF-BD cases and compared with unselected FMF and BD controls. RESULTS: The prevalence of BD was higher in FMF than in populations known to be rich in BD (eg, 16 per 4,000 in FMF compared with 1 per 104 in Japan, P < .001). FMF-BD cases and FMF or BD controls were comparable in most demographic, clinical, and laboratory aspects studied. However, more cases than FMF-controls were of Iraqi/Turkish origin and responded less favorably to colchicine. A higher proportion of cases than BD controls had skin, central nervous system, and gastrointestinal manifestations, originated from North Africa, and had family history of BD. In most cases, as in most respective controls, the severity of FMF was of intermediate grade and the extensiveness of BD was limited. The HLA B5 antigen was present in 53% of BD cases and 40% of BD controls. CONCLUSIONS: BD should be included among the vasculitides complicating FMF. BD and FMF in patients with FMF-BD, and in patients suffering from each of these entities alone, are clinically and demographically comparable. PMID- 10805354 TI - Clinical evolution, and morbidity and mortality of primary Sjogren's syndrome. AB - OBJECTIVES: To study the clinical and laboratory profile evolution, as well as morbidity and mortality impact, of primary Sjogren's syndrome (pSS), in a large cohort of patients followed-up longitudinally. METHODS: We studied the evolution of the clinical picture and laboratory profile of pSS, the incidence and predictors for systemic sequelae, and the impact of pSS on overall survival in a prospective cohort study of 261 patients with pSS. Analyses included calculation of incidence rates, Cox proportional hazards predictive models, and estimation of standardized mortality ratios (SMRs) compared with the general Greek population, adjusting for age and sex. RESULTS: Glandular manifestations of the syndrome were typically present at the time of diagnosis. Systemic manifestations such as arthritis, Raynaud's phenomenon, purpura, interstitial nephritis, and liver involvement, as well as the serological profile, also did not change substantially during subsequent follow-up. Incidence rates for peripheral neuropathy, glomerulonephritis, and lymphoproliferative disorders were 3.3, 6.6, and 12.2 per 1,000 person-years, respectively. Glomerulonephritis and lymphoma tended to co-exist in the same patients (relative risk, 34.0; P < .0001). The development of lymphoproliferative disorders was associated with low levels of C4 complement (relative risk, 7.5; P = .0016), the presence of mixed monoclonal cryoglobulins (relative risk, 7.9; P = .0012), and purpura (relative risk, 3.9; P = .037). Low levels of C4 was the strongest predictor for mortality after adjusting for age (relative risk, 6.5; P =.0041). Patients with pSS had an SMR of 2.07 (95% CI, 1.03 to 3.71). However, when patients with adverse predictors were excluded, the mortality rate was identical to that of the general population (SMR 1.02). CONCLUSIONS: The initial presentation of pSS determines subsequent outcome. Purpura, decreased C4 complement levels, and mixed monoclonal cryoglobulinemia are adverse prognostic factors. The overall mortality of patients with pSS compared with the general population is increased only in patients with adverse predictors. PMID- 10805355 TI - The costs of rheumatoid arthritis: an international long-term view. AB - OBJECTIVES: To review the literature on the measurable direct and indirect costs of rheumatoid arthritis (RA) in industrialized countries from a societal perspective and to develop a template for international use. METHODS: A literature search using MEDLINE and other sources identified 153 relevant published articles, press releases, and so forth on the costs of RA and rheumatism from the major Organization for Economic Cooperation and Development (OECD) countries in English and other languages. Sixty-eight publications provide some economic data for analysis and are included in the bibliography. Twelve publications provide sufficiently detailed and robust information for inclusion in country overview tables. The concept of varied costs at different disease stages measured by years since diagnosis and Health Assessment Questionnaire (HAQ) scores is used to guide rational decisions in the allocation of scarce health care resources. RESULTS: Direct costs increase overproportionately during the course of the disease. The most important driver of direct costs is hospitalization, especially in moderate and severe RA. Costs of medication represent a comparatively small proportion of direct costs. Indirect costs caused by work disability can be substantially higher than direct costs, particularly in working-age patients. The total costs of RA to society, and the different cost components such as direct and indirect costs, are broadly comparable in industrialized countries by their order of magnitude. Major confounding factors for international comparison are different study methodologies and patient samples. CONCLUSIONS: The cost template developed in this article can be used to estimate the likely costs of RA to society for industrialized countries. It probably will underestimate indirect costs because of their incomplete coverage in the studies examined. A long-term perspective is needed for chronic diseases such as RA to assess the future effects of early interventions. Treatment in the early stages of RA that effectively reduces long-term disability has the potential to save substantial costs to society. PMID- 10805356 TI - Intravenous immunoglobulin treatment of lupus nephritis. AB - OBJECTIVE: To evaluate the clinical response of treatment-resistant membranous and membranoproliferative lupus nephritis to intravenous immunoglobulin (IVIg). METHODS: Seven lupus nephritis patients who failed to respond to at least prednisone and cyclophosphamide were studied. A kidney biopsy showing either membranous or membranoproliferative glomerulonephritis was available in six patients. They were treated with six courses (patients 1 and 2) or 1 or 2 courses (patients 3 through 7) of high-dose IVIg. For patients 3 through 7, the plasma levels of albumin, total cholesterol, urea, creatinine, dsDNA antibody titers, and daily proteinuria were measured just before the IVIg therapy, immediately on completion, and 6 months later. RESULTS: All seven patients had a beneficial response to IVIg. In patient 1, decrease in proteinuria was evident 2 weeks after IVIg was started, nephrotic syndrome gradually disappeared, and she had no proteinuria in 3 years' follow-up. Decline in proteinuria was evident in patient 2 after the 4th IVIg course, but proteinuria reached the pretreatment level 4 months after the therapy ended. In patients 3 through 7, the mean daily proteinuria before IVIg (5.3 +/- 2.1 g) decreased after 1 or 2 IVIg courses (3.3 +/- 1.4 g), and further decreased when measured 6 months later (2.1 +/- 1.3 g). Similarly, the plasma cholesterol level decreased while the plasma albumin level increased after IVIg. CONCLUSIONS: IVIg might be effective in treatment-resistant membranous or membranoproliferative lupus nephritis. Future studies should concentrate on determining the preferred treatment protocol of IVIg for the various classes of lupus nephritis. PMID- 10805357 TI - Anti-dsDNA antibodies in sarcoidosis. AB - BACKGROUND: Sarcoidosis is a chronic multisystem disorder characterized by an exaggerated cellular immune response to antigens with the production of various antibodies including rheumatoid factor and antinuclear antibodies (ANA). The prevalence and significance of antibodies to double-stranded DNA (anti-dsDNA) in sarcoid patients is unknown. The occurrence of anti-dsDNA antibodies is known to be a specific marker of systemic lupus erythematosus (SLE). Sarcoidosis can occur with SLE. It is unclear if anti-dsDNA antibodies in patients with sarcoidosis signify the eventual development of SLE. OBJECTIVES: To determine the prevalence of anti-dsDNA antibodies in patients with sarcoidosis in a university hospital and their significance in predicting the diagnosis of associated SLE. METHODS: In a retrospective study, 34 patient files with diagnosed sarcoidosis in a university hospital during a period of 15 years were reviewed for serological markers, including ANA, anti-dsDNA, and immunoglobulin and C3 levels. The occurrence of SLE in these patients also was evaluated. RESULTS: ANA were positive in 10 of 34 of the patients screened. Two patients with sarcoidosis had antibodies to dsDNA. C3 levels in these 34 patients were an average of 87.7 +/- 25.3 mg/100 mL, which is within the normal range. IgG immunoglobulin levels were an average of 2,206 +/- 999 mg/100 mL, which was above normal limits. The 2 patients who were positive for anti-dsDNA had normal C3 levels and SLE did not develop during a follow-up period of 10 to 15 years. CONCLUSIONS: Anti-dsDNA antibodies may occur in patients with sarcoidosis, but their presence does not predict the subsequent development of SLE. PMID- 10805358 TI - Potential therapeutic approach to SAPHO. PMID- 10805359 TI - Bacterial vaginosis: the quest continues. PMID- 10805360 TI - Images in infectious diseases in obstetrics and gynecology. Lethal sepsis due to traditional healing. PMID- 10805361 TI - Prevalence and clinical significance of postpartum endometritis and wound infection. AB - OBJECTIVE: To correlate clinical variables (gestational age, severe pregnancy induced hypertension, gestational diabetes mellitus, history of previous cesarean sections, fetal distress, perinatal mortality, postpartum anemia, Apgar score < or = 3 at 1 minute and < or = 7 at 5 minutes, and instrumental delivery) with postpartum endometritis (PPE) and wound infection. METHODS: Descriptive cross sectional study of the outcome of 75,947 term and preterm singleton deliveries; vaginally and by cesarean section from 1989-1997. RESULTS: The prevalence of PPE after vaginal deliveries was 0.17% (120/68,273). Gestational age of less than 37 weeks, severe pregnancy-induced hypertension, fetal distress, instrumental deliveries, neonatal mortality, postpartum anemia, and Apgar scores of < 7 after 5 minutes were significantly associated with PPE. Gestational diabetes and an Apgar score of < 3 after 1 minute showed similar frequency with and without PPE. The prevalence of PPE after cesarean section was 2.63% (202/7,677). Preterm cesarean sections, history of previous cesarean sections, anemia, and low Apgar scores were seen more frequently with PPE than without. The incidence of cesarean delivery with gestational diabetes mellitus, fetal distress, and perinatal mortality was similar in presence and absence of PPE. The rate of wound infection after cesarean section was 3.97% (318/7,995). Gestational diabetes mellitus, history of previous cesarean deliveries, and low Apgar scores were significantly more frequent with than without wound infection. Gestational age, severe pregnancy-induced hypertension, fetal distress, perinatal mortality, and postpartum anemia were not associated with wound infection. CONCLUSIONS: Awareness of the aforementioned associations may prevent and shorten hospital stay by early diagnosis and appropriate treatment. PMID- 10805363 TI - Risk factors for a complicated clinical course among women hospitalized with pelvic inflammatory disease. AB - The aim of this study was to identify factors ascertainable at initial presentation that predict a complicated clinical course in HIV-negative women hospitalized with pelvic inflammatory disease (PID). We used data from a cross sectional study of women admitted for clinically diagnosed PID to a public hospital in New York City. A complicated clinical course was defined as undergoing surgery, being readmitted for PID, or having a prolonged hospital stay (> or = 14 days) but no surgery. Logistic regression was used to identify independent predictors of complications. In adjusted analyses, older age (> or = 35 years) was a risk factor for prolonged hospital stay (adjusted odds ratio [OR] = 3.9; 95% confidence interval [CI] = 1.3-11.6) and surgery (OR = 10.4; CI = 2.5 44.1); self-reported drug use was a risk factor for readmission for PID (OR = 7.7; CI = 1.4-41.1) and surgery (OR = 6.2; CI = 1.8-20.5). Older age and self reported drug use appear to be independent risk factors for a complicated clinical course among women hospitalized with PID. PMID- 10805362 TI - Association of lower genital tract inflammation with objective evidence of endometritis. AB - The purpose of this report is to evaluate the association between lower genital tract inflammation and objectively diagnosed endometritis. We analyzed the first 157 patients enrolled in the PEACH study, a multicenter randomized clinical trial designed to compare the effectiveness of outpatient and inpatient therapy for PID. Women less than 38 years of age, who presented with a history of pelvic discomfort for 30 days or less and who were found to have pelvic organ tenderness (uterine or adnexal tenderness) on bimanual examination, were initially invited to participate. After recruitment of the first 58 patients (group 1) we added the presence of leukorrhea, mucopurulent cervicitis, or untreated positive test for N. gonorrhoeae or C. trachomatis to the inclusion criteria (group 2, N = 99). We compared rates of endometritis in the two groups and calculated the sensitivity, specificity, and predicted values of the presence of white blood cells in the vaginal wet preparation. The rate of upper genital tract infection in group 1 was 46.5% (27/58) compared to 49.5% (49/99) in group 2. Microbiologic evidence of either N. gonorrhoeae or C. trachomatis increased from 22.4% in group 1 to 38.3% in group 2. The presence of vaginal white blood cells or mucopus has a high sensitivity (88.9%), but a low specificity (19.4%) for the diagnosis of upper genital-tract infection. Assessment of the lower genital tract for evidence of infection or inflammation is a valuable component of the diagnostic evaluation of pelvic inflammatory disease. The presence of either mucopus or vaginal white blood cells is a highly sensitive test for endometritis in patients with pelvic pain and tenderness. PMID- 10805364 TI - Association between HIV in pregnancy and antiretroviral therapy, including protease inhibitors and low birth weight infants. AB - OBJECTIVE: To determine the incidence of low birth weight infants born to HIV seropositive women and to demonstrate any effects of antiretroviral therapy on birth weight. METHODS: Retrospective review of all obstetrical medical records from January 1, 1995 through June 30, 1998 to identify HIV seropositive women. We evaluated their antiretroviral therapy, CD4 counts, and birth weights of their newborns. We conducted detailed review of the clinical and laboratory findings for the HIV-infected untreated patients, women who received ZDV antepartum alone, and those who received PIs as part of antiretroviral treatment. RESULTS: The frequency of low birth weight infants was significantly increased in HIV seropositive compared to HIV seronegative parturients. Low birth weight infants were more frequent among HIV infected women with lower CD4 counts but the association was not statistically significant. Women who received no antepartum treatment, antepartum only ZDV, and those treated with PIs had significantly more low birth weight infants than did comparison groups. HIV seropositive women also had high frequencies of several obstetrical risk factors for low birth weight infants. CONCLUSION: The present study showed a significantly increased frequency of low birth weight infants among HIV infected women and especially the subgroups of infected women who received no antepartum treatment, antepartum ZDV only, and those treated with PIs. This association, however, may be related to the presence of many other preterm obstetrical risk factors noted in this study. Increasing numbers of HIV seropositive women are being treated with PIs according to the Centers for Disease Control (CDC) guidelines. If PIs are a cause of low birth weight infants, women taking these drugs may have incremental risk of low birth weight. PMID- 10805365 TI - In vitro inhibition of commercial douche products against vaginal microflora. AB - Recently, vaginal douching has been associated with many health risks in women. The aim of this study was to analyze the effect of commercial douche products against various vaginal microorganisms, including lactobacilli. Seven commercial douches were tested against eight Lactobacillus clinical isolates and three type strains from the American Type Culture Collection. BV-associated bacteria included six strains of five genera: Gardnerella, Mobiluncus, Mycoplasma, Peptostreptococcus, and Ureaplasma. Two isolates of group B Streptococcus, and three species of Candida were also tested. The minimal inhibition concentrations and minimal contact times for these products against vaginal microorganisms were determined in broth cultures. Four antiseptic-containing douche products showed a strong inhibitory effect against all vaginal microorganisms tested with a short contact time (less than 1 min). Three vinegar-containing douche products selectively inhibited vaginal pathogens associated with bacterial vaginosis, group B streptococcal vaginitis, and candidiasis, but not lactobacilli. The antimicrobial effects of the commercial douche products varied among different brands and microbial species tested. PMID- 10805366 TI - Effect of imiquimod on cytokine induction in first trimester trophoblasts. AB - OBJECTIVES: Imiquimod (IQ) is used clinically for the topical treatment of external genital warts. IQ is an immune response modifier and induces the expression of interferon-alpha and other cytokines in human Peripheral Blood Monocytes (PBMC). Trophoblasts have been previously shown to express inflammatory cytokines upon lipopolysaccharide (LPS) stimulation. The objective of this study was to evaluate the ability of IQ to induce transcription of cytokines in trophoblasts. METHODS: A transformed human first trimester trophoblast cell line, HTR-8/SVneo, was cultured in DMEM containing IQ at concentrations of 0 to 5.0 microg/ml. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) viability assays were conducted to control for any drug-induced cell death. Total RNA was isolated from trophoblasts at 0, 8 and 24 hours of culture and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was conducted using specific amplimers for the inflammatory cytokines interleukin (IL)-1alpha, IL-1beta, IL-6 and IL-8. RT-PCR of beta-actin was performed to control for equal RNA loading. RESULTS: RT-PCR was unable detect an increase in either IL-1alpha, IL-1beta, IL-6 or IL-8 mRNA in first trimester trophoblasts cultured in the presence of 0 to 5.0 microg/mL of IQ for up to 24 hours. RT-PCR confirmed equal RNA loading and MTT viability assays did not show loss of cell viability at concentrations of IQ up to 5.0 microg/ml. CONCLUSIONS: IQ, at the concentrations tested, did not induce the transcriptional expression of inflammatory cytokines in human first trimester trophoblasts. These data suggest that IQ would not induce the expression of inflammatory cytokines in placental trophoblasts. PMID- 10805367 TI - Comparison of Abbott LCx Chlamydia trachomatis assay with Gen-Probe PACE2 and culture. AB - In this study the LCx assay (a nucleic acid amplification assay) for Chlamydia trachomatis in endocervical samples was compared with the Gen-Probe PACE2 assay (a nucleic acid probe assay) for endocervical samples, and with endocervical culture. In addition, the efficacy of the LCx assay was determined for midstream clean-catch urine samples because it is often necessary to obtain such a sample for routine urine culture and it is simpler to collect only a single sample without also collecting a first-void urine for LCx. Endocervical specimens from 205 patients were tested for C. trachomatis via LCx and PACE2. Of these patients, 203 were tested by culture. Midstream clean-catch urine samples from 75 of these patients were tested by LCx. The sensitivities and specificities for these assays, after discrepant analysis, were 100 and 98.9% for LCx of endocervical samples, 52.4 and 100% for PACE2; and 71.4 and 100% for culture. The sensitivity/specificity of LCx for midstream clean-catch urines was 66.7/98.5%. The apparent prevalence of C. trachomatis in our population was 10.2%. These data indicate that among the methods tested, LCx of endocervical samples had the highest sensitivity for C. trachomatis in this population. The sensitivity of the urine LCx assay using midstream clean-catch collected urines was considerably less than that reported in other studies that used first-void urines but was higher than that of PACE2. PMID- 10805368 TI - Regression of engineered tumor cells secreting cytokines is related to a shift in host cytokine profile from type 2 to type 1. AB - The precise role of the endogenous immune response in modulating cancer development remains unclear. In this study, three mouse tumor cell lines were used to elucidate the immune mechanisms for tumor regression versus tumor growth. These cell lines were (1) the poorly immunogenic VKCK cell line and (2) its two derived cell lines VKCK/RM4-tumor necrosis factor-alpha (TNF-alpha) and VKCK/RM4 interferon-gamma (IFN-gamma) engineered to secrete TNF-alpha and IFN-gamma, respectively. Our data showed that VKCK tumors grew aggressively in syngeneic BALB/c mice, and vaccination of irradiated VKCK cells failed to protect the mice from a subsequent challenge with the same tumor. In contrast, engineered VKCK tumor cells lost their tumorigenicity, and vaccination of engineered VKCK cells induced a protective immunity against VKCK cells that was mediated with VKCK specific CD8+T cells. Susceptible mice developed a Th2-dominant response, whereas resistant mice developed a Th1-dominant response to VKCK. The T cell proliferative response and cytolytic activity against VKCK developed in both resistant and susceptible mice, but in the susceptible mice, these responses were much weaker compared with those in the resistant mice. Our results indicate that regression of tumor cells engineered to secrete cytokines TNF-alpha and IFN-gamma is related to a shift from a host type 2 to a type 1 cytokine profile. Our results further suggest that the failure of unmodified VKCK to generate efficacious T cells is not due to an inability to recognize tumor antigens but, rather, to the nature and magnitude of the antitumor immune response that develops. A better understanding of the mechanisms by which tumor cells modulate the host immune system may result in newer approaches for manipulating host-tumor interactions that favor the development of a protective antitumor immune response. PMID- 10805369 TI - Type I interferons mediate the lipopolysaccharide induction of macrophage cyclin D2. AB - Lipopolysaccharide (LPS) is a powerful macrophage-activating agent and antimitogen. We recently showed that LPS unexpectedly induces cyclin D2 in macrophages. Since LPS stimulates macrophages to produce autocrine-acting cytokines, we examined whether LPS induction of cyclin D2 was mediated by one such type of cytokine, type I interferons (IFN). We report that bone marrow derived macrophages (BMM) lacking a component of the type I interferon receptor (IFNAR-1) do not express cyclin D2 mRNA or protein in response to LPS stimulation (0.01-1 microg/ml for 7-30 h). Consistent with this result, addition of anti-IFN alpha/beta neutralizing antibodies reduced levels of LPS-stimulated cyclin D2 in normal BMM. Furthermore, IFN-alpha alone induced cyclin D2 mRNA and protein in normal BMM. Thus, we have identified a new role for type I IFN in macrophages, namely, as essential mediators of LPS-stimulated cyclin D2 expression. PMID- 10805370 TI - Endotoxin and mast cell granule proteases synergistically activate human coronary artery endothelial cells to generate interleukin-6 and interleukin-8. AB - Mast cells (MC) are strategically located along blood vessels and, on activation, exocytose granules that contain many vasoactive mediators. Endothelial cell (EC) activation, which includes the production of such cytokines as interleukin-6 (IL 6) and IL-8, is a key event in vascular inflammation. In this study, the effects of purified MC granules (MCG) on the production of IL-6 and IL-8 by human coronary artery EC (HCAEC) were examined. HCAEC were cocultured with MCG in the presence or absence of lipopolysaccharide (LPS), and IL-6 and IL-8 levels in the culture medium were assayed by ELISA. Unactivated HCAEC produced only low levels of IL-6 or IL-8, and the addition of MCG alone resulted in little or no increase in production of these cytokines. LPS-activated HCAEC produced significant amounts of IL-6 and IL-8 in a dose-dependent and time-dependent fashion, which was amplified 2-3-fold by MCG at EC/MC ratios of 16:1-2:1. Scanning electron microscopy revealed direct communication between MCG and HCAEC. The enhancement of IL-6 and IL-8 production by MCG was abrogated when MCG were pretreated with the serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF). These results demonstrate that MCG interaction with HCAEC causes amplification of endotoxin stimulated cytokine production via serine proteases present in MCG. The synergistic activation of EC by endotoxin and MCG proteases emphasizes the role of MC in amplifying vascular inflammation. PMID- 10805371 TI - Tumor necrosis factor-alpha and lipopolysaccharide enhance interferon-induced antichlamydial indoleamine dioxygenase activity independently. AB - In macrophages, interleukin-1 (IL-1) and lipopolysaccharide (LPS) enhance the antichlamydial effect of interferon-gamma (IFN-gamma) by increasing indoleamine 2,3-dioxygenase (IDO) activity in a dose-dependent manner. Our objectives were to characterize the antichlamydial effect of tumor necrosis factor-alpha (TNF-alpha) on IFN-induced IDO activity and to establish the relationship between LPS and TNF alpha in IDO potentiation. TNF-alpha inhibited chlamydial growth in a dose dependent manner only in IFN-treated macrophages. Furthermore, excess tryptophan reversed the effect of combined cytokine treatment, indicating that IDO alone was responsible for chlamydial inhibition. The promonocyte THP-1 cell line, previously used to model the effect of IL-1 on IDO mRNA expression, was treated with IFN-gamma and increasing concentrations of LPS or TNF-alpha. IDO mRNA was quantified by RT-PCR, and IDO activity was measured by HPLC at 24 and 48 h after treatment, respectively. Both LPS and TNF-alpha enhanced IDO activity and IDO mRNA expression, with maximal IDO induction at 100 ng/ml LPS or 5 ng/ml TNF alpha. Anti-TNF-alpha failed to neutralize the effects of LPS treatment, and insufficient TNF-alpha or IL-1 was produced by LPS-treated THP-1 cells to account for the enhancing effect of LPS, indicating that the effect of LPS on IDO was independent of TNF-alpha and IL-1. PMID- 10805372 TI - The action of tumor necrosis factor-alpha on rat mast cells. AB - Taking into account that cytokine tumor necrosis factor-alpha (TNF-alpha) and mast cells (MC) both are involved in inflammation, it seems of great importance to recognize their relationships. Therefore, we have studied whether recombinant human TNF-alpha (rHuTNF-alpha) can cause histamine secretion from rat peritoneal MC. We have also examined the effect of this cytokine on MC reactivity. We have established that TNF-alpha stimulates rat MC to histamine release in a concentration-dependent manner. TNF-alpha-induced histamine secretion was evoked by concentrations > 10-16 M and reached the maximum rate at a concentration of 10 10 M (histamine release 17.1% +/- 1.9%, mean +/- SEM). We have also noticed that pretreatment of MC with TNF-alpha (in a concentration of 10-16 M) significantly inhibited concanavalin A (ConA)-stimulated release of histamine, with the percent release decreasing to 51% of the control value. Treatment of mast cells with TNF alpha resulted in a decrease of compound 48/80-dependent histamine release as well (the percent released histamine fell to 85% of the control value). This altered MC responsiveness was reversible. After 120 min of resting time, the MC reactivity came back to the initial values. We have concluded that TNF-alpha appears to be a direct stimulus for MC to release histamine, and it may regulate MC secretory function. PMID- 10805373 TI - Interferon-beta induces selective enhancement of antigen-specific T cell responses. AB - Interferon-beta (IFN-beta) inhibits mitogen-induced T cell responses, in part through downregulation of interleukin-12 (IL-12) or upregulation of IL-10. We have reexamined these findings using ragweed (RW) stimulated or tetanus toxoid (TT)-stimulated human peripheral blood mononuclear cells (PBMC) and nontransformed, antigen-specific, human Th0, Th1, and Th2 clones. IFN-beta induced concentration-dependent inhibition of phytohemagglutinin (PHA)-stimulated PBMC proliferation and enhancement of RW-stimulated or TTstimulated PBMC proliferation. Monocyte depletion of PBMC isolates resulted in concentration dependent inhibition of RW-driven or TT-driven proliferation by IFN-beta. This response was unaltered by the addition of either exogenous recombinant human IL 12 (rHuIL-12) or saturating concentrations of anti-IL-10. Moreover, addition of exogenous rHuIL-10 to nondepleted RW-driven or TT-driven PBMC cultures did not alter the concentration-dependent enhancement of antigen-driven proliferation induced by IFN-beta. Th0, Th1, and Th2 clones stimulated in the presence of antigen and autologous, irradiated PBMC displayed concentration-dependent inhibition of proliferation in the presence of IFN-beta that was unaltered by the addition of either exogenous rHuIL-12 or a saturating concentration of anti-IL 10. Finally, whereas IFN-beta inhibited antigen-driven generation of IL-5, IL-12, IL-13, and IFN-gamma, IFN-beta enhanced generation of both IL-4 and IL-10. Thus, IFN-beta, induces a selective, IL-10-independent and IL-12-independent upregulation of antigen-specific T cell responses, supporting the role of IFN beta as an immunomodulatory rather than an antiproliferative/immunosuppressive cytokine. PMID- 10805374 TI - Secretion of CXC chemokine IP-10 by peripheral blood mononuclear cells from healthy donors and breast cancer patients stimulated with HER-2 peptides. AB - CXC chemokines play an important role in recruitment of T cells to the site of activation and regulation of angiogenesis. CXC chemokines are secreted by T cells stimulated with cytokines or by established cytotoxic T lymphocyte (CTL) lines at recognition of conventional antigen (Ag), but the activation requirements and the relationship of interferon-gamma (IFN-gamma) inducible protein (IP-10) secretion with IFN-gamma induction in lymphocytes are still unclear. We studied the induction of IP-10 from nonadherent peripheral blood mononuclear cells (PBMC) by IFN-gamma, interleukin-12 (IL-12), and the HER-2 peptide E75, which forms a CTL defined antigen. We found that IFN-gamma alone was a weak inducer of IP-10 in these cells, whereas IL-12 was a significantly stronger inducer of IP-10. In the presence of IL-12, the tumor peptide E75 (HER-2, 369-377) was a stronger inducer of IP-10 than was IL-12 alone. E75 and its variants mutated at position 5 could also induce IP-10 in the absence of exogenous IL-12 or IFN-gamma. IP-10 induction by E75 required HLA-A2 presentation and B7-CD28 interactions and was partially inhibited by blocking of CD40-CD40L interactions. These results indicate that presentation of tumor peptides to peripheral T cells can induce a fast chemokine response, which in its early phase may be higher than the IFN-gamma response. This shows that the IP-10 response was independent of any early-phase IFN-gamma response in peripheral T cells. This may be important for understanding the regulation of the balance between chemoattractant chemokines (CC) and CXC chemokines by tumor Ag and may have implications for understanding the mechanisms of polarization of T cells and conditioning of antigen-presenting cells (APC) by tumor antigens. PMID- 10805375 TI - Expression of IFN-gamma in cerebrovascular endothelial cells from aged mice. AB - Recently, it has become clear that interferon-gamma (IFN-gamma) plays a role in the central nervous system (CNS) as well as in the immune system. However, the reason for the alteration in IFN-gamma production in the brain with aging remains unknown. In this study, we investigated the expression of IFN-gamma in the brain in terms of both mRNA and protein and compared the expression in young adult brain with that in aged mice. The cerebrum and cerebellum were collected from young adult (8-10 weeks old) and aged (24-26 months old) BALB/c mice, and the expressions of IFN-gamma and IFN-gamma receptor-1 (IFNGR-1) mRNA were examined by RT-PCR. Expression of IFN-gamma mRNA was detected in the brains from aged mice but not in those from young adult mice. However, IFNGR-1 mRNA was expressed in the brains from both young adult and aged mice. Moreover, IFN-gamma levels in the cerebrum and cerebellum from aged mice were detectable by ELISA, but IFN-gamma was undetectable in these tissues from young adult mice. To identify the cellular source of IFN-gamma in the brain of aged mice, immunostaining using antimouse IFN gamma monoclonal antibody (mAb) was done. Immunoreactivity of IFN-gamma appeared to be located in cerebrovascular endothelial cells, including the choroid plexus of the cerebellum from aged mice. Expression of IFN-gamma and IFNGR-1 was also identified in isolated microvessels from brains. These results suggest that IFN gamma plays a role in age-associated changes. PMID- 10805376 TI - Inhibition of interferon, cytokine, and lymphocyte proliferative responses in elite swimmers with altitude exposure. AB - To determine the immunologic consequences of athletic training at altitude, blood samples were taken at rest from 10 swimmers and 8 control nontraining but altitude-exposed members of the 1996 Australian Olympic Swimming Team, near the start and completion of a 21-day training camp at 2102 m. Blood leukocyte numbers dropped in both groups (p < 0.05), with the decrease greater in the swimmers ( 38% swimmers, -3% controls). Concanavalin A (ConA)-induced blastogenesis decreased in both groups (p < 0.01), but the drop was greater in the control group (-32% swimmers, -56% controls, p < 0.05). Lipopolysaccharide (LPS)-induced blastogenesis more than doubled in both groups (281% swimmers, 249% controls, p < 0.01). Increases in mitogen-induced interleukin-1beta (IL-1beta), IL-4, and interferon-gamma (IFN-gamma) production and a decrease in IL-2 levels were observed in both groups after altitude exposure (all p < 0.05). The percentage of cells expressing HLA-DR fell (-33% swimmers, -20% controls, p < 0.01), whereas those expressing CD-4 expression increased (16% swimmers only, p < 0.01). Although training at medium-level altitude alters some immunologic parameters, the training-induced changes may be secondary to those induced by altitude alone. PMID- 10805377 TI - The expression and biologic effects of ovine interleukin-4 on T and B cell proliferation. AB - Using the reverse-transcriptase polymerase chain reaction (RT-PCR), cDNA encoding ovine (Ov) interleukin-4 (OvIL-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (PBMC). Two identical clones generated from separate RT-PCR reactions differed from a published OvIL-4 sequence, although they had a high degree of identity with the bovine and human homologs. We show by sequence analysis that the OvIL-4 cDNA retained the four alpha-helix structure and disulfide bonds identified in human IL-4 (HuIL-4). Moreover, the cDNA encoding OvIL-4 was expressed in insect cells using the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) as a vector. Supernatants from insect cells infected with the recombinant virus secreted an additional protein with a relative molecular mass of 17,000. This protein was recognized by an anticervine IL-4 monoclonal antibody (mAb) in a Western blot and did not react with any proteins in supernatants from uninfected insect cells or cells infected with the wild-type AcMNPV. Supernatants from insect cells infected with the recombinant virus induced the proliferation of activated B cells in a dose dependent manner and typically demonstrated 5 x 105 dilution U/ml of activity. However, OvIL-4 had no effect on the proliferation of resting T cells isolated from efferent lymph and actually inhibited the ability of a mitogen to stimulate these resting lymphocytes. In contrast, OvIL-4 induced the proliferation of mitogen-activated lymphoblast, demonstrating the complex role(s) OvIL-4 plays in the regulation of B and T cells. PMID- 10805378 TI - Localization and post-Golgi trafficking of tumor necrosis factor-alpha in macrophages. AB - Tumor necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine secreted by activated macrophages. In this study, we examined the intracellular distribution and trafficking of TNF-alpha. Immunofluorescence and immunogold localization demonstrated that in lipopolysaccharide (LPS)-stimulated RAW264 macrophages, the greatest concentration of TNF-alpha is found in the perinuclear Golgi complex. Staining of the Golgi complex appeared 20 min after activation of cells and persisted for 2-12 h, and TNF-alpha appeared on the cell surface only transiently during this time. The rate of disappearance of Golgi staining correlated with the release of the cleaved, mature TNF-alpha into the medium. Pulse chase labeling and subcellular fractionation studies indicated that both 26-kDa and 17-kDa forms of TNF-alpha may be present at the level of the Golgi complex. Post-Golgi trafficking of TNF-alpha was modulated by agents that disrupt the cytoskeleton. Interferon-gamma (IFN-gamma), which primes macrophages for TNF-alpha-dependent cellular cytotoxicity, potentiated the effect of LPS by sustaining enhanced intracellular pools of TNF-alpha and also promoted redistribution of TNF-alpha into post-Golgi vesicular compartments. We propose that the primary pool of biologically active TNF-alpha in activated macrophages is held in the Golgi complex and that the cytokine is recruited directly from this intracellular pool for release in response to tumor cells or pathogens. PMID- 10805379 TI - Eleventh International Congress on Transfer Factors: March 1-4, 1999, Monterrey, Nuevo Leon, Mexico. PMID- 10805380 TI - Double staining of intracellular cytokine proteins and T-lymphocyte subsets. Evaluation of the method in blood and bronchoalveolar lavage fluid. AB - An immunocytochemical staining method has been developed for simultaneous staining of both cell surface markers (CD4 and CD8) and intracellular cytokine proteins IFN-gamma, IL-4 and IL-5. Cell surface molecules were visualized with alkaline phosphatase, which was developed by Fast Blue BB. Intracellular cytokine proteins were detected by amino-ethyl carbazole. We applied this technique to T cells from T-cell lines and T-cell clones, peripheral blood mononuclear cells and broncho-alveolar lavage fluid cells. Cells were used either unstimulated or stimulated for 4 h with 1 ng/ml PMA and 1 microg/ml ionomycin, which proved to be an optimal stimulus taking cytokine staining, cell recovery and cell viability into account. We studied peripheral blood mononuclear cells from healthy subjects and found that without in vitro stimulation on average 0.4% of the cells were IFN gamma positive cells. In unstimulated broncho-alveolar lavage fluid cells of the 2 allergic asthmatic subjects studied so far we found higher numbers of cytokine positive cells (up to 22% of the lymphocytes being IL-4+ cells). By in vitro stimulation, the numbers of cytokine-positive peripheral blood mononuclear cells from the healthy subjects were increased to maximally 5% IFN-gamma+ cells. In stimulated lavage fluid cells from allergic asthmatic subjects maximally 34% of the lymphocytes became IFN-gamma+. We conclude that this method allows detection of intracellular cytokine proteins in both CD4+ and CD8+ T cells without the need for stimulating the cells in vitro. In vitro stimulation may change the cytokine profile detected. PMID- 10805382 TI - Visualization of carbohydrate-binding molecules expressed by myelomono- and erythropoietic cells derived from human bone marrow: an immunoenzymatic double staining study. AB - Interactions between human haematopoietic and bone marrow stromal cells are governed by complex carbohydrate-mediated adhesion processes. In order to evaluate corresponding carbohydrate-binding sites on human myelo- and erythropoietic cells which were able to react with mono- or oligosaccharides, we established an immunocytochemical double-staining assay. In a first step, cell lineages were visualized using Lewis(x) (CD15) or glycophorin C-specific monoclonal antibodies. The second step included polyacrylamide-conjugated carbohydrate structures. According to our results, the carbohydrate-binding potential of granulopoietic cells increased during the process of maturation, contrasting a reduction of carbohydrate-binding sites on erythroid precursor cells during differentiation. With respect to previous in vitro studies, these findings shed some light on certain aspects of bone marrow homing as well as on the trafficking of mature cellular elements into circulation. It is tempting to speculate that carbohydrate-mediated adhesion mechanisms may be involved in the various functional defects of progenitor cells in chronic myelogenic leukaemia, especially regarding their complex interactions with the marrow microenvironment. PMID- 10805381 TI - A role for SSeCKS, a major protein kinase C substrate with tumour suppressor activity, in cytoskeletal architecture, formation of migratory processes, and cell migration during embryogenesis. AB - SSeCKS is a major protein kinase C substrate which has tumour suppressor activity in models of src- and ras-induced oncogenic transformation. The mitogenic regulatory activity of SSeCKS is likely manifested by its ability to bind key signalling proteins such as protein kinases C and A and calmodulin, and to control actin-based cytoskeletal architecture. Rat SSeCKS shares extensive homology with human Gravin, an autoantigen in myasthenia gravis that encodes kinase scaffolding functions and whose expression pattern in fibroblasts and nerves suggests a role in cell motility. Here, we analyse the expression of SSeCKS and Gravin in rodent and human fibroblast and epithelial cell lines using antibodies specific or crossreactive for SSeCKS or Gravin. SSeCKS expression was then analysed in developing mouse embryos and in adult tissues. In the foetal mouse, early SSeCKS protein expression (E10-11) is focused in the loose mesenchyme, luminal surface of the neural tube, notochord, early heart and pericardium, urogenital ridge, and dorsal and ventral sections of limb buds. In later stages (E12-14), SSeCKS is widely expressed in mesenchymal cells but is absent in the spinal ganglia. By E15, SSeCKS expression is ubiquitous, although the staining pattern varies from being striated within smooth muscle sarcomeres to filamentous in mesenchymal and select epithelial cells. In the adult mouse, SSeCKS staining is relatively ubiquitous, with highest expression in the gonads, smooth and cardiac muscle, lung, brain and heart. High expression is also detected in fibroblasts and nerve fibres as well as in more specialized cells such as glomerular mesangial cells and testicular Sertoli cells. SSeCKS expression in the rat testes correlates with the induction of puberty, and in mature mouse spermatozoa, SSeCKS is found in peripheral acrosome membranes and in a helix-like winding pattern within the midsection. Periodic enrichments of SSeCKS are found in sperm midsections and in developing axons, suggesting a role in architectural infrastructure. As with Gravin, high SSeCKS expression is absent in most epithelial cells; however, in contrast to Gravin, SSeCKS is expressed in Purkinje cells, cardiac muscle, macrophages and hepatic stellate cells, indicating overlapping yet distinct patterns of tissue expression in the SSeCKS/Gravin family. The data suggest roles for SSeCKS in the control of cytoskeletal and tissue architecture, formation of migratory processes and cell migration during embryogenesis. PMID- 10805383 TI - Characterisation of lectin binding patterns of mouse bronchiolar and rat alveolar epithelial cells in culture. AB - Lung epithelial cell differentiation pathways remain unclear. This is due in part to the plasticity of these cells and the lack of markers which accurately reflect their differentiation status. The aim of this study was to determine if lectin binding properties are useful determinants of functional differentiation status in vitro. Mouse Clara cells were cultured for 5 days. During this time, no alteration in differentiation was evident by electron microscopy. No significant alteration in binding reactivity of Bauhinia purpurea (BPA), Maclura pomifera (MPA), Concanavalin A, Wheat germ or Helix pomatia lectins occurred in cultures compared with Clara cells in mouse lung tissue. In contrast, nitrotetrazolium blue reductase activity and CC10 expression declined in culture. Rat type II cells were cultured for 8 days. Between days 0 and 4, the number of type II cells identified by electron microscopy was constant at 70-80%, decreasing to 8% by day 6. In contrast, by day 4, only 42% cells retained alkaline phosphatase activity. BPA and MPA reactivity was altered at day 0 and day 4 respectively, compared with cells in situ. Therefore, the reactivity of lectins analysed here does not reflect functional differentiation status of cultured mouse Clara cells. However, BPA and MPA reactivity may be a sensitive indicator of alterations in rat type II cell differentiation in vitro. PMID- 10805385 TI - Immunocytochemical localization of taurine in different muscle cell types of the dog and rat. AB - The presence and distribution of the amino acid taurine in different muscle cell types of the dog and rat was examined by immunocytochemical methods. The light microscope study revealed that smooth muscle cells were similarly immunoreactive for taurine, whereas skeletal muscle fibres showed wide differences in taurine immunoreactivity among individual cells. Some skeletal fibres were strongly immunoreactive whereas others did not display immunolabelling. Mononucleated satellite cells, found adjacent to skeletal fibres in a quiescent stage, were also immunostained. Other myoid cells, such as testicular peritubular cells showed a cytoplasmic and a nuclear pool of taurine. By means of electron microscope immunolabelling, the subcellular localization of taurine was studied in vascular and visceral smooth muscle cells. Taurine was present in most subcellular compartments and frequently appeared randomly distributed. Taurine was localized on myofilaments, dense bodies, mitochondria, the plasma membrane and the cell nucleus. Moreover, the labelling density within individual smooth muscle cells was variable and depended on the state of contraction of each single fibre. Contracted cells showed a higher density of gold particles than relaxed cells. Unmyelinated nerve fibres, found adjacent to smooth muscle cells from the muscularis mucosae and the lamina propria of the stomach, were unstained or poorly stained. PMID- 10805384 TI - Histochemical evidence for inducible nitric oxide synthase in advanced but non ruptured human atherosclerotic carotid arteries. AB - In response to cytokine stimulation, the inducible isoform of the nitric oxide synthase (iNOS) produces large amounts of nitric oxide with potential consequences in the pathophysiology of atherosclerosis. Previous investigations have demonstrated the presence of iNOS in human atherosclerotic lesions. The goal of this study was to evaluate the occurrence of the expression of iNOS in ruptured versus non-ruptured human carotid atherosclerotic plaques. Using plastic embedded sections, we performed in situ hybridization and immunohistochemistry on very advanced atherosclerotic lesions type V (non-ruptured) and type VI (ruptured) from 12 atheromatous carotid arteries from endarterectomy and six non atherosclerotic internal mammary arteries from aorto-coronary bypass. Only one internal mammary artery expressed iNOS in the endothelium. In contrast, iNOS mRNA and protein were repeatedly expressed in advanced lesions type V in 5/7 cases, particularly in inflammatory regions. Specific cell markers identified iNOS positive cells as macrophages and T-lymphocytes but also as smooth muscle cells and endothelial cells adjacent to these inflammatory regions. Nitration of protein tyrosines was not always associated to iNOS expression but more likely to the presence of inflammatory cells. In complicated lesions type VI, the occurrence of iNOS mRNA and protein expression diminished drastically (1/5 cases). Combined expression of iNOS mRNA and protein is frequently found in advanced but non-ruptured human atherosclerotic carotid lesions while it becomes rare after the plaque has ruptured. These findings suggest that iNOS could be an active participant in the plaque rupture event. PMID- 10805387 TI - Artefacts in iodine ion milling of some compound semiconductors AB - Iodine ion milling was developed to overcome the deficiencies of using argon as a milling gas for certain compound semiconductors. Here it is shown that iodine gas introduces strong artefacts in some materials such as Zn3As2, a material occurring at some ZnSe/GaAs interfaces and that even efficient specimen cooling does not prevent this effect. Argon ion milling by contrast leaves the Zn3As2 layer intact. However, deliberate amorphisation of the Zn3As2 by iodine enables a clearer detection of its presence and to distinguish better the coverage of very thin ZnSe epilayers. These effects can be accounted for by simple thermodynamic calculations and enable the possibility for prediction as to whether reactive milling agents such as iodine will cause problems in specimen preparation. These results also indicate that iodine molecules rather than atoms are the principle milling species. PMID- 10805386 TI - The nervous system of the chicken proventriculus: an immunocytochemical and ultrastructural study. AB - The proventriculus constitutes the glandular region of the chicken stomach. This organ is innervated by two parasympathetic networks, the myenteric and submucous plexus, and here we present a systematic study of this system by immunohistochemistry and electron microscopy. All the neurons and fibres were positive for the neural markers, protein gene product 9.5 and the amidating enzymes. Immunoreactivities for the constitutive neuronal isoform of the enzyme nitric oxide synthase and the vasoactive intestinal peptide were present in neuronal bodies suggesting an intrinsic origin for the similarly immunoreactive fibres found in the proventriculus. On the other hand, immunoreactivity to gastric inhibitory peptide was only found in varicose fibres making contact with the blood vessels and the glandular epithelium, but never in the neuronal somas, suggesting that this substance may be provided by an extrinsic nervous system whose neuronal bodies are located elsewhere. Electron microscopy revealed frequent neuromuscular and neuroepithelial connections in the muscle layers, the wall of the blood vessels and the epithelium. In addition, synapsis-like structures were identified in the proximity of cells belonging to the diffuse endocrine system, providing a new example of neuroendocrine contacts. No positivity was found for antibodies against other neural substances including somatostatin, peptide histidine-isoleucine, peptide tyrosine-tyrosine, neuropeptide tyrosine, bombesin, met-enkephalin, serotonin, substance P, galanin, calcitonin gene-related peptide and S-100 protein. PMID- 10805388 TI - The orientation-dependent simulation of ELNES AB - We describe a program that allows the simulation of energy-loss near edge structure (ELNES). As an extension to the WIEN97 package (a full potential linearized augmented plane wave package for calculating crystal properties) [1] it permits to separate different contributions to the inelastic scattering cross section according to the character of the final state, explicitly taking into account projection onto scattering vector and integration over collection and convergence angle. Thus the program facilitates analysis of ELNES under precisely defined experimental conditions, and allows the investigation of anisotropic effects in ELNES from crystal structures. Dipole-allowed as well as dipole forbidden transitions can be analyzed with this program. PMID- 10805389 TI - Shear force distance control in near-field optical microscopy: experimental evidence of the frictional probe-sample interaction AB - The properties of the probe-surface contact for a near-field optical microscope driven in the shear force mode have been studied applying lateral amplitudes of the probing fiber tip larger than 15 nm. Electric current measurements between a conductive tip and a conductive sample reveal a pulsed current behavior at the very beginning of the approach curve. In the upper part of the approach curve it turns to the quasiconstant current. From this observation a conclusion is drawn about the presence of permanent mechanical contact between the probe and the surface in the shear force mode. A shift of the approach curve along the z-axis as a function of dither amplitude was discovered. These results are in contradiction to the established conception of possible physical mechanisms of shear force interaction. To settle this issue the friction model is proposed according to which the damping of the probe vibrations is caused by the friction between the tip and the surface. PMID- 10805390 TI - A storage Dewar near-field scanning optical microscope AB - A near-field scanning optical microscope for operation within a storage Dewar is described. It was designed for studies of opaque samples and operates in the collection mode. Illumination can be either through the tip or from the side via a separate fiber. Scans can be begun within 2 h after start of cooldown. Its rigid design allows high resolution and long scans with no additional vibration isolation. To illustrate its performance, measurements of photoluminescence in GaAs/AlGaAs heterostructures are presented. The signal and noise levels for the two illumination modes are examined. PMID- 10805391 TI - Comparison of electron diffraction data from non-linear optically active organic DMABC crystals obtained at 100 and 300 kV AB - During the recent past, we have synthesized a new class of molecules with intramolecular two-dimensional charge transfer upon excitation. The present report presents such a molecule, 2,6-bis(4-dimethylamino-benzylidene) cyclohexanone (DMABC), with an unusually high value of the second-order non linear optical (NLO) coefficients. In order to optimize the macroscopic NLO properties of the compounds, it is necessary to relate their first hyperpolarizability tensors at a molecular level to those at a crystal bulk level. This requires a complete structure determination and refinement. However, the growth of sufficiently large single crystals, which are needed for structural analysis and refinement by X-ray methods, is a time consuming and sometimes impossible task. We have performed a complete structural analysis by electron diffraction combined with simulation methods and with maximum entropy and log likelihood statistics. In order to improve the quantitative analysis, a 300 kV data set using an on-line CCD camera was added and the best attainable R-values were compared with those from 100 kV data using film emulsions. Details regarding the maximum attainable resolution for both data sets are discussed as well as the problems which arise from the limited dynamic range in photographic emulsions as compared to a 14 bit CCD camera. Once the crystal structure was known, quantum chemical methods were used to calculate non-linear optical susceptibility tensor components and these were related to the macroscopic coefficients of the crystalline quadratic non-linearity tensor. In the present work, both ab initio and semi-empirical quantum-chemical calculations were employed. PMID- 10805392 TI - Preparation of damage-free glass TEM specimens AB - A jet polishing technique to chemically thin glass specimens for transmission electron microscopy (TEM) after a preliminary mechanical dimpling step has been developed. Slightly modified commercial equipment is used with automatic optical termination of the polishing process to produce foils exhibiting large, high quality, electron transparent regions. PMID- 10805393 TI - Quantitative phase-sensitive imaging in a transmission electron microscope AB - This paper presents a new technique for forming quantitative phase and amplitude electron images applicable to a conventional transmission electron microscope. With magnetised cobalt microstructures used as a test object, we use electron holography to obtain an independent measurement of the phase shift. After a suitable calibration of the microscope, we obtain quantitative agreement of the phase shift imposed on the 200 keV electrons passing through the sample. PMID- 10805394 TI - Incoherent imaging with the soft X-ray microscope AB - The imaging characteristics for X-ray wavelengths in the "water window" under incoherent imaging conditions (large detector aperture) are examined for the King's College London scanning transmission X-ray microscope with zone-plate objective installed at the Daresbury (UK) synchrotron. The principal consideration was to express image theory, incorporating wave aberrations and apodised zone plates, and to apply the theory to experimental data. Comparisons are made, showing reasonable agreement, for a range of defocus values and two wavelengths. Due to problems in fabrication it was necessary to determine the effective, or operational, zone-plate parameters (radius of outermost active zone rN, width of outermost active zone drN); this was accomplished by through-focus series. Calculated point spread functions were used to deblurr images, in through focus series of two-dimensional scanned X-ray images of specimen holes and test grating patterns. Significant contrast enhancement is achieved after deconvolution with a best point-to-point resolution of about 35 nm. PMID- 10805396 TI - A practical simulated annealing program and its application to quantitative CBED pattern matching AB - A modified simulated annealing (SA) program is presented (source code available from the authors). The program was used for the refinement procedure in quantitative convergent beam diffraction (QCBED). Considering the practical situation in QCBED, three modifications have been made to the conventional SA algorithm. First, a most probable search boundary (MPB) was suggested, which guarantees the program to search mainly within the MPB, but allows it also to go out of the boundary sometimes. Secondly, the MPB was set to shrink during the minimization. This algorithm decreases the computing time significantly. Thirdly, a more meaningful stop criterion was adopted using the nominal physical precision of each refinement parameter. The feasibility and the efficiency of the modified program were tested using both calculated and experimental CBED patterns. The presented program is proved to be capable of high performance. PMID- 10805395 TI - International telemicroscopy with a 3 MV ultrahigh voltage electron microscope. AB - The ability for remote microscope operation via a network connection was added recently to the ultrahigh voltage electron microscope (UHVEM) in Osaka University, and used successfully for the observation of thick biological samples across the Pacific Ocean by researchers at the National Center for Microscopy and Imaging Research (NCMIR) at the University of California San Diego. High-quality images at video rate were transferred by a satellite link and control signals were transmitted by an ISDN connecting the workstations at both sites. Most microscope functions operated from the console of the UHVEM were replicated on the graphical user interface of the remote workstation. By clicking on icons or in boxes in the display window with a mouse, the researcher could operate the UHVEM from the remote-site. The total delay time for sending images and returning control signals was about 0.7 s, which did not interfere significantly with the smooth operation of the instrument. Researchers at the remote site were able to record images on film in the microscope which were later sent to San Diego. PMID- 10805397 TI - Impact of column bending in high-resolution transmission electron microscopy on the strain evaluation of GaAs/InAs/GaAs heterostructures AB - The accuracy of strain profiles obtained by a quantitative analysis of lattice fringe spacings from high-resolution micrographs is discussed. Focusing on highly lattice mismatched GaAs/InAs/GaAs heterostructures the local strain distribution of the layers is calculated by finite element simulations to determine the atom positions in elastically relaxed transmission electron microscopy specimens. By analysing simulated images a significant decoupling between the layer structure and the contrast pattern motifs is found for relevant imaging conditions, which may result in an incorrect determination of strain profiles and layer compositions when examining experimental micrographs. PMID- 10805398 TI - Charging phenomena in PEEM imaging and spectroscopy. AB - Spectromicroscopy with the imaging technique of X-ray photoelectron emission microscopy (X-PEEM) is a microchemical analytical tool installed in many synchrotron radiation laboratories, and which is finding application in diverse fields of research. The method of sample analysis, X-ray absorption spectroscopy, does not encounter the same problems as X-ray photoemission spectroscopy when sample charging occurs, hence even good insulators may often be analyzed without any apparent artifacts in images or spectra. We show, however, that charging effects cannot be neglected. We model the effect of surface charge formation on the secondary electron yield from uniform samples to demonstrate that surface charge primarily reduces the yield of electrons which may contribute to the detected signal. We illustrate that on non-uniform insulating samples, localized centers of charge may substantially affect microscope imaging and resolution as the electrostatic field close to the surface is distorted. Finally, in certain circumstances non-uniform surface charge may lead to unexpected lineshapes in X ray absorption spectra causing, in some extreme cases, negative spectra. These negative spectra are explained, and several strategies are reviewed to minimize the impact of sample charging when analyzing poorly conducting samples of any nature. PMID- 10805399 TI - Role of endothelium and nitric oxide in experimental hypertension. AB - A short review on the role of endothelium and nitric oxide (NO) in experimental hypertension is presented in the light of the literature and our own recent findings. Based on these data, it is concluded that even though there is a lot of evidence in favor of the primary and causal association of endothelial dysfunction and NO in experimental hypertension, it seems still more plausible that they are causative in some types of hypertension only. Our own experience rather speaks for a secondary but still an important participation of endothelium in the maintenance and further elevation of high blood pressure. Endothelium plays a key role in the development of organ damages in hypertension. PMID- 10805400 TI - Age-related endothelial dysfunction with respect to nitric oxide, endothelium derived hyperpolarizing factor and cyclooxygenase products. AB - Vascular aging is associated with both structural and functional changes that can take place at the level of the endothelium, vascular smooth muscle cells and the extracellular matrix of blood vessels. With regard to the endothelium, reduced vasodilatation in response to agonists occurs in large conduit arteries as well as in resistance arteries with aging. Reviews concerning the different hypotheses that may account for this endothelial dysfunction have pointed out alterations in the equilibrium between endothelium-derived relaxing and constricting factors. Thus, a decreased vasorelaxation due to nitric oxide and, in some arteries, endothelium-derived hyperpolarizing factor as well as an increased vasoconstriction mediated by cyclooxygenase products such as thromboxane A2 are likely to occur in age-induced impairment of endothelial vasodilatation. Furthermore, enhanced oxidative stress plays a critical role in the deleterious effect of aging on the endothelium by means of nitric oxide breakdown due to reactive oxygen species. The relative contribution of the above phenomenon in age related endothelial dysfunction is highly dependent on the species and type of vascular bed. PMID- 10805401 TI - Inducible NO synthase activity in blood vessels and heart: new insight into cell origin and consequences. AB - Induction of the inducible form of nitric oxide synthase (iNOS) in the vascular and cardiac tissue by several inflammatory stimuli may result in the production of large amounts of nitric oxide (NO) for a sustained period. Recent data obtained in the rat aorta in which iNOS was induced by lipopolysaccharide (LPS) have demonstrated that adventitial cells represent the main site of NO production. Adventitial-derived NO can exert an immediate down-regulatory effect on smooth muscle contraction (via activation of the cyclic GMP pathway) but may also initiate longer lasting effects through the formation of NO stores within the medial layer. One candidate for such NO stores are dinitrosyl non-heme iron complexes. Low molecular weight thiols interact with preformed NO stores and promote vasorelaxation by a cyclic GMP-independent mechanism involving the activation of potassium channels. In the heart, the induction of iNOS is involved in delayed protection against ischemia-reperfusion-induced functional damages. Recent data obtained with monophosphoryl lipid A, a non-toxin derivative of LPS, strongly suggest that iNOS-derived NO in the rat heart does not act as an immediate mediator of the cardioprotection but rather as a trigger of long-term protective mechanisms. Thus, the present data reveal the important role of adventitial cells as a site of iNOS expression and activity in intact blood vessels. The induction of adaptive mechanisms in the heart and the formation of releasable NO stores in blood vessels are examples of long-term consequences of iNOS induction. These new information are relevant for a better understanding of the circumstances in which NO overproduction by iNOS may play either a beneficial or deleterious role in these tissues. PMID- 10805402 TI - Nitric oxide-compromised hypertension: facts and enigmas. AB - NO concentration in the femoral artery and femoral vein of anesthetized dogs was found to be 154.2+/-5.6 nM and 90.0+/-12 nM, respectively. Inhibition of NO synthase (NOS) slightly decreased the basal NO concentration in femoral artery from 154.2+/-5.6 to 137.2+/-3.3 nM. Acetylcholine-induced increase in NO concentration was slightly but still significantly attenuated, suggesting that very probably L-NAME did not inhibit all sources of nitric oxide (NO). Local NOS inhibition in the posterior hypothalamus dose-dependently increased systemic blood pressure (BP) in rats. Short-term general NOS inhibition in anesthetized dogs increased diastolic BP but not systolic BP. The heart rate after one-hour down-fluctuation returned to initial values. Proteosynthesis in the myocardium and both branches of the left coronary artery increased, but this was not supported by polyamines, since the activity of ornithine decarboxylase declined. Long-term general NOS inhibition elicited a sustained BP increase, a decrease in heart rate, cardiac hypertrophy and an increase in wall thickness of the coronary and carotid artery. The results indicate that NO deficiency itself plays a role in proteosynthesis and cardiac hypertrophy, in spite of relatively small increase in diastolic blood pressure and no change in systolic blood pressure, at least after an acute L-NAME administration. The hypotension response to acetylcholine and bradykinin studied in anesthetized NO-compromised rats, was unexpectedly enhanced. The elucidation of this paradoxical phenomenon will require further experiments. PMID- 10805403 TI - The potential role of nitric oxide in the hypertrophic growth of the left ventricle. AB - Left ventricular hypertrophy (LVH) is the result of interaction between a chronic hemodynamic overload and non-hemodynamic factors. There are several lines of evidence presented in this work suggesting that nitric oxide (NO) may participate in the hypertrophic growth of the myocardium. First, endothelial NO production was shown to be decreased in several types of hemodynamically overloaded circulation both in animals and humans. Second, compounds stimulating NO production were able to diminish the extent or modify the nature of LVH in some models of myocardial hypertrophic growth. Third, arterial hypertension can be induced by inhibition of nitric oxide synthase activity. This NO-deficient hypertension is associated with the development of concentric LVH, myocardial fibrosis and protein remodeling of the left ventricle. The mechanism of LVH development in NO-deficient hypertension is complex and involves decreased NO production and increased activation of the renin-angiotensin-aldosterone system. Cardiovascular protection via ACE inhibition in NO-deficient hypertension may be induced by mechanisms not involving an improvement of NO production. In conclusion, the hypertrophic growth of the LV appears to be the result of interaction of vasoconstrictive and growth stimulating effects of angiotensin II on the one hand and of vasodilating and antiproliferative effects of nitric oxide on the other. PMID- 10805404 TI - Long-term administration of D-NAME induces hemodynamic and structural changes in the cardiovascular system. AB - N(G)-nitro-D-arginine-methyl ester (D-NAME) is considered to be an inactive enantiomer of L-NAME and is generally used as the negative control for NO synthase inhibition with L-NAME. With the aim to compare the effects of 4-week L NAME and D-NAME treatments on hemodynamic and cardiovascular structural parameters, four groups of male Wistar rats were investigated: the controls and groups administered 40 and 20 mg/kg/day of L-NAME and 40 mg/kg/day of D-NAME. At the end of the experiment, myocardial NO synthase activity decreased by 42, 24 and 25%; aortic NO synthase activity decreased by 35, 15 and 13% vs. controls in the L-NAME 40, L-NAME 20 and D-NAME 40 groups, respectively. The DNA concentrations in the myocardium and the aorta increased significantly after L NAME and D-NAME treatments. The inhibition of NO synthase was accompanied by a significant elevation in systolic blood pressure in all three groups. The LVW/BW ratio increased by 27, 14 and 13% vs. controls in the L-NAME 40, L-NAME 20 and D NAME 40 groups, respectively. The aortic wall mass, measured as the cross sectional area, increased by 45, 17 and 25% vs. controls in the L-NAME 40, L-NAME 20 and D-NAME 40 groups, respectively. Myocardial fibrosis represented 0.94% in the controls, but 7.96, 4.70 and 5.25% in L-NAME 40, L-NAME 20 and D-NAME 40 groups, respectively. It is concluded that D-NAME, although less affective than L NAME, inhibits NO synthase activity resulting in hemodynamic and structural changes in the cardiovascular system similar to the changes induced by half the dose of L-NAME. Thus, the consideration of D-NAME as an inactive enantiomer and its use as the negative control needs to be reevaluated. PMID- 10805405 TI - Effect of captopril on cyclic nucleotide concentrations during long-term NO synthase inhibition. AB - The aim of the present study was to determine the effect of angiotensin converting enzyme inhibitor captopril on cGMP and cAMP concentration in the left ventricle and aorta after NO synthase inhibition by 4-week-lasting N(G)-nitro-L arginine-methyl ester (L-NAME) treatment. Five groups of rats were investigated: controls, L-NAME in the dose 20 mg/kg/day (L-NAME 20), L-NAME in the dose 40 mg/kg/day (L-NAME 40), captopril in the dose 100 mg/kg/day, L-NAME 40 mg/kg/day together with captopril 100 mg/kg/day. Captopril completely prevented L-NAME induced hypertension and LV hypertrophy development. Compared to the controls, cGMP concentration in the L-NAME 20 and L-NAME 40 groups was decreased by 13% and 22%, respectively, in the left ventricle and by 27% and 56% in the aorta, respectively. Captopril did not influence this decrease of cGMP concentration. Cyclic AMP concentration in the aorta of L-NAME 20 group increased by 17%. In the L-NAME 40 group, cAMP concentration increased by 17% in the left ventricle and by 34% in the aorta compared to controls. This increase was enhanced in rats given L NAME together with captopril. Captopril alone had no effect on cAMP concentration. We conclude that captopril does not affect the concentration of cGMP, however, it has more than the additive effect on the cAMP concentration increase in the cardiovascular system during long-term NO synthase inhibition. PMID- 10805406 TI - Short-term NO synthase inhibition and the Na+-binding properties of cardiac Na,K ATPase. AB - It is known that hypertension is accompanied by increased [Na+]i. The functional properties of Na,K-ATPase, which transports the Na+ out and K+ into myocardial cells during the relaxation phase, were investigated in the left ventricle (LV), septum (SV) and the right ventricle (RV) of anesthetized dogs with moderate acute blood pressure elevation elicited by short-term (4-hour) NO synthase inhibition. The NO-insufficiency was induced by administration of an L-arginine analogue, the N(G)-nitro-L-arginine methyl ester (L-NAME). Concerning the function of Na,K ATPase under the conditions of lowered NO synthesis, we focused our attention to the binding of Na+ to the enzyme molecule. Activation of the enzyme by increasing Na+ concentrations revealed significant changes in both the maximal velocity (Vmax) and the affinity for Na+ (K(Na)) in all investigated heart sections. The Vmax increased by 27% in LV, by 87% in SV and by 58% in RV. The K(Na) value increased by 86% in LV, by 105% in SV and by 93% in RV, indicating an apparent decrease in the sensitivity of the Na+-binding site in the Na,K-ATPase molecule. This apparently decreased pump affinity for Na+ together with the increase of Vmax suggest that, during the short-term inhibition of NO synthesis, the Na,K ATPase is capable of extruding the excessive Na+ from the myocardial cells more effectively at higher [Na+]i, as compared to the Na,K-ATPase of control animals. PMID- 10805407 TI - Induction of angiogenesis in NO-deficient rat heart. AB - Angiogenesis is known to be triggered by various stimuli including hypertension. It was previously found that NO-deficient hypertension is accompanied by structural remodeling of the cardiac muscle and large coronary arteries. This study was aimed to examine the qualitative subcellular alterations of capillaries in the heart of the rats treated with L-NAME (40 mg/kg/day for 4 weeks). The results showed that long-lasting inhibition of NO production induced an apparent activation of fibroblast function. This was associated with enhancement of fibrotization as well as with the induction of angiogenesis. Accordingly, fibroblasts were frequently located in the vicinity of capillary pericytes, which was followed by their detachment and migration. Moreover, besides inactive or even injured capillaries, the other ones exhibited extensive proteosynthetic activity linked to capillary growth, proliferation and migration of endothelial cells. The results strongly indicate enhanced triggering of the angiogenesis in L NAME-induced NO-deficient hypertension. PMID- 10805408 TI - Chronic disturbances in NO production results in histochemical and subcellular alterations of the rat heart. AB - The mechanisms and myocardial alterations associated with NO-deficient hypertension are still far from clear. The aim of the present study was to focus on the enzyme histochemical and subcellular changes in the heart of L-NAME treated rats, as well as to examine the influence of captopril treatment. Wistar rats were administered either L-NAME (40 mg/kg/day) alone or together with captopril (100 mg/kg/day) for a period of 4 weeks. A significant increase of blood pressure confirmed the reliability of the model. The results showed that long-lasting L-NAME administration was accompanied by a decrease of endothelial NO-synthase activity and by a significant local decrease of the following enzyme activities: capillary-related alkaline phosphatase, 5'-nucleotidase and ATPase (but not dipeptidyl peptidase IV) and cardiomyocyte-related glycogen phosphorylase, succinic dehydrogenase, beta-hydroxybutyrate dehydrogenase and ATPases. No activity of these enzymes was found in the scar, whereas a marked increase of alkaline phosphatase and dipeptidyl peptidase IV activities was found in the foci of fibrotization. Histochemical changes correlated with subcellular changes, which were characterized by 1) apparent fibroblast activation associated with interstitial/perivascular fibrosis, 2) heterogeneous population of the normal, hypertrophic and injured cardiomyocytes, 3) enhancement of the atrial granules and their translocation into the sarcolemma, and 4) impairment of capillaries as well as by induction of angiogenesis. Similar alterations were also found in the heart of captopril co-treated rats, despite of the significant suppression of blood pressure. The results indicate that NO-deficient hypertension is accompanied by metabolic disturbances and ultrastructural alterations of the heart and these changes are probably not induced by the renin angiotension system only. PMID- 10805410 TI - Cross-talk between nitric oxide and HSP70 in the antihypotensive effect of adaptation to heat. AB - In this work, we evaluated the effect of adaptation to heat on the fall of blood pressure (BP) induced by heat shock (HS) and the interrelation between nitric oxide (NO) and heat shock protein, HSP70. Experiments were carried out on Wistar rats. It was shown that HS resulted in a generalized and transient increase in NO production (the electron paramagnetic resonance method) and a fall of BP from 113+/-3 to 88+/-1 mm Hg (p<0.05). Adaptation to heat itself did not affect BP, but completely prevented the NO overproduction and hypotension induced by HS. The adaptation simultaneously increased the brain NO-synthase content and induced HSP70 synthesis (the Western blot analysis) in various organs. Both the antihypotensive effects of adaptation and HSP70 accumulation were completely prevented by L-NNA, an inhibitor of NO synthesis, or quercetin, an inhibitor of HSP70 synthesis. The data suggest that adaptation to heat stimulates NO synthesis and NO activates synthesis of HSP70. HSP70, which hampers NO overproduction, thus restricts the BP fall induced by heat shock. PMID- 10805409 TI - Role of nitric oxide in adaptation to hypoxia and adaptive defense. AB - Adaptation to hypoxia is beneficial in cardiovascular pathology related to NO shortage or overproduction. However, the question about the influence of adaptation to hypoxia on NO metabolism has remained open. The present work was aimed at the relationship between processes of NO production and storage during adaptation to hypoxia and the possible protective significance of these processes. Rats were adapted to intermittent hypobaric hypoxia in an altitude chamber. NO production was determined by plasma nitrite/nitrate level. Vascular NO stores were evaluated by relaxation of the isolated aorta to diethyldithiocarbamate. Experimental myocardial infarction was used as a model of NO overproduction; stroke-prone spontaneously hypertensive rats (SHR-SP) were used as a model of NO shortage. During adaptation to hypoxia, the plasma nitrite/nitrate level progressively increased and was correlated with the increase in NO stores. Adaptation to hypoxia prevented the excessive endothelium dependent relaxation and hypotension characteristic for myocardial infarction. At the same time, the adaptation attenuated the increase in blood pressure and prevented the impairment of endothelium-dependent relaxation in SHR-SP. The data suggest that NO stores induced by adaptation to hypoxia can either bind excessive NO to protect the organism against NO overproduction or provide a NO reserve to be used in NO deficiency. PMID- 10805412 TI - Contemporary activation of different endothelial receptors accounts for a reserve mechanism of nitric oxide-mediated relaxation. AB - The aim of this study was to investigate whether the inhibition of one of the endothelial receptor sites in the rat pulmonary artery (muscarinic, histaminergic, purinergic, alpha2-adrenergic) affects the NO-mediated relaxation induced by the activation of the other type of receptors. Acetylcholine (ACh)-, histamine (Hist)-, adenosine (Ade)-, and clonidine (Clon)-induced endothelium dependent relaxations were reduced by the administration of specific antagonists of muscarinic, H1-histaminergic, purinergic or alpha2-adrenergic receptors, respectively. The inhibition of H1-histaminergic receptors by chlorphenyramine did not prevent ACh-induced relaxation. Similarly, the inhibition of muscarinic receptors by atropine did not prevent the relaxations to histamine, adenosine and clonidine. On the other hand, the relaxations induced by acetylcholine, histamine, adenosine or clonidine were regularly reduced by NO-synthase inhibitor N(G)-nitro-L-arginine methyl ester (10(-4) mol/l). These results suggest that the inhibition of NO-synthase abolished arterial relaxations induced by all agonists. After inhibition of one type of the endothelial receptors, the NO-dependent relaxation could still be evoked by activation of one of the others. PMID- 10805411 TI - Histamine-induced relaxation in pulmonary artery of normotensive and hypertensive rats: relative contribution of prostanoids, nitric oxide and hyperpolarization. AB - The aim of this study was to determine the relative contribution of nitric oxide (NO), endothelium-derived hyperpolarizing factor (EDHF) and prostanoids in histamine-induced relaxation of isolated pulmonary artery from normotensive and hypertensive rats. The hypertension was induced by oral administration of NO synthase inhibitor N(G)-nitro-L-arginine methylester (L-NAME, 50 mg/kg/day) to normotensive rats for 8 weeks. In phenylephrine-precontracted arterial rings the histamine-induced relaxation was significantly reduced in L-NAME-treated rats compared to the controls. Indomethacin (cyclooxygenase inhibitor) and glibenclamide (ATP-sensitive K+-channel blocker) did not inhibit the relaxation response in either control or hypertensive rats. On the other hand, tetraethylammonium (TEA), a K+-channel blocker with a broad specificity, significantly reduced histamine-induced relaxation in the pulmonary artery from both groups examined. The TEA-resistant relaxation was completely abolished by additional administration of L-NAME to the incubation medium. The results indicate that histamine-induced relaxation of the pulmonary artery in both normotensive and hypertensive rats is mediated mainly by nitric oxide, whereas EDHF seems to play a minor role. PMID- 10805414 TI - Effect of indomethacin and deendothelisation on vascular responses in the renal artery. AB - Vasodilator prostaglandins (PGE2, PGI2) play an important role in the regulation of renal blood flow. Hence, inhibition of their production with nonsteroidal anti inflammatory drugs increases renal vascular resistance and exerts adverse renal effects. It has been reported that besides endothelium-derived prostaglandin products, nitric oxide (NO) may be mainly involved in regulation of renal functions. The aim of our study was to evaluate the effect of cyclooxygenase inhibition with indomethacin and endothelium removal on vascular responses of the renal artery as a model vessel. Isolated segments of rabbit renal arteries were perfused at constant flow. Indomethacin administration (10(-5) mol x l(-1)) significantly increased the responses to single doses (0.1, 1, 10 microg) of noradrenaline (NA) as compared with the controls. In indomethacin-pretreated vessels, subsequent deendothelisation by air bubbles enhanced the constrictor responses to NA. In reversed order, when deendothelisation was followed by indomethacin administration, the responses to NA were similar in character. A comparison of renal artery responses to NA in both experimental situations did not reveal any significant differences. It can be supposed that endothelial and non-endothelial factors may be involved in local regulation of renal vascular tone. PMID- 10805413 TI - Evaluation of endothelium-protective effects of drugs in experimental models of endothelial damage. AB - Endothelium-protective properties of pharmacological agents may be assessed by using different experimental models of endothelial dysfunction or injury. The model of endothelial dysfunction induced by vessel perfusion with polymorphonuclear leukocytes (PMN) was used for evaluation of pentoxifylline (PTX) effects on vasoconstrictor responses to noradrenaline (NA) in the rabbit renal artery. Addition of PMN into the perfusion solution significantly increased the responses to NA at all doses. PTX administration (10(-5) mol x l(-1)) significantly diminished the constrictor responses to NA in vessels perfused with PMN+PTX when compared to the responses in PMN-perfused vessels (at dose 0.1 microg: 32.25 vs. 14.25, at dose 1 microg: 51 vs. 27.75 (p<0.01), at dose 10 microg 74.25 vs. 39.75 (p<0.05), all values expressed as median of perfusion pressure in mm Hg). The model of endothelial damage induced by repeated NA administration in 5 doses (10-50 microg of NA) was used for evaluation of the endothelium-protective effect of sulodexide (SLX). It was found that SLX (120 U/l) significantly decreased the number of desquamated endothelial cells (EC) compared to the control group (controls: 131.4+/-20.1 EC, +SLX: 83.3+/-13.8 EC, p<0.01). These results confirmed the favorable endothelium-protective effects of pentoxifylline and sulodexide in the two experimental models. PMID- 10805415 TI - Passive smoking impairs endothelium-dependent relaxation of isolated rabbit arteries. AB - The aim of the present study was to examine the effect of prolonged passive smoking (lasting 3 weeks) on plasma catecholamine levels and reactivity of isolated rabbit arteries. Plasma noradrenaline, adrenaline and dopamine levels were determined radioenzymatically. Isolated rings of the thoracic aorta and carotid artery were suspended in organ chambers and connected to a force transducer for the recording of isometric tension. Plasma noradrenaline levels were found to be significantly elevated in rabbits subjected to passive smoking for 3 weeks. Plasma adrenaline and dopamine levels were not changed. Transmural nerve stimulation of arterial rings evoked frequency-dependent contractions. Prolonged passive smoking did not affect neurogenic contractions of the arteries tested. On the other hand, endothelium-dependent relaxations of phenylephrine precontracted arteries were significantly impaired. Furthermore, hypertrophy of the left ventricle was observed. In conclusion, passive smoking impairs endothelium-dependent relaxations but not neurogenic contractions of systemic arteries. The impaired relaxations of arteries may be, at least in part, mediated through the degradation of released nitric oxide by superoxide anions derived from cigarette smoke. PMID- 10805416 TI - Inhibition of NO synthase activity in nervous tissue leads to decreased motor activity in the rat. AB - The nitric oxide/cGMP system has been shown to play a crucial role in the mechanism of learning and memory. The aim of the present study was to investigate whether the inhibition of NO synthase in brain regions leads to alterations of spontaneous behavior in rats. Male Wistar rats were treated with NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) at the dose of 40 mg/kg/day. After 4 weeks of L-NAME treatment, NO synthase activity was significantly decreased by 75% in the cerebellum, by 71% in the cerebral cortex and by 72% in the thoracic spinal cord. Decreased NO synthase activity in the nervous tissue was associated with decreased motor horizontal and vertical activities as well as by lowered frequency of sniffing, cleaning and defecation. It is concluded that the inhibition of NO synthase activity has a suppressive effect on spontaneous behavior of rats. PMID- 10805417 TI - Partial colocalization of NADPH-diaphorase and acetylcholinesterase positivity in spinal cord neurons. AB - The freely diffusible radical, nitric oxide (NO), has been assumed to act as a retrograde signaling molecule that modulates transmitter release. Acetylcholine (ACh) is known to function as a typical neurotransmitter. In the present work we have examined the presence of both transmitters (NO and ACh) and their possible relations in the rabbit spinal cord. In our experiments we have used histochemical methods for the visualization of acetylcholinesterase (AChE) and NADPH diaphorase (NADPH-d) which label neurons that express nitric oxide synthase (NOS). Both histochemical methods were performed separately or together on the same sections of the thoracic spinal cord. NADPH-d positive dark blue stained neurons were seen mostly in superficial and deep layers of the dorsal horn, preganglionic autonomic neurons and pericentral area. The presence of AChE positive amber yellow neurons was confirmed mostly in motoneurons located in the ventral horns and in neurons of the pericentral and intermediate zone. Besides the above mentioned neurons, also double-labeled neurons were found which contained both the yellow and dark blue histochemical product. Their presence was confirmed in the intermediate zone and in the pericentral area. Thus, the co existence of NADPH-d and AChE occurred in the location of interneurons. Our observations suggest that NO may play a role in the control of cholinergic neuronal activity and that NO can be involved in the modulation of synaptic transmission. PMID- 10805418 TI - Short-term changes of NADPH diaphorase-exhibiting neuronal pools in the spinal cord of rabbit after repeated sublethal ischemia. AB - The aim of this study was the histochemical characterization of NADPH diaphorase positive neuronal pools in the rabbit lumbosacral segments using a model of single, repeated and multiple sublethal spinal cord ischemia. Following a single 8-min sublethal spinal cord ischemia and 1-hour reperfusion, the staining of NADPH diaphorase-exhibiting neurons in the dorsal horn, pericentral region, dorsal gray commissure and sacral parasympathetic nucleus was comparable with the control sections. In contrast to the foregoing sublethal ischemia, a regionally different somatic NADPH diaphorase (NADPHd) staining was found after multiple sublethal spinal cord ischemia. Whereas an almost complete loss of the staining of large NADPHd-exhibiting somata in the pericentral region was detected, the staining of the NADPHd-exhibiting neuronal pools in the deep dorsal horn and sacral parasympathetic nucleus was fully preserved. Concomitantly, a prominent reduction of small NADPH diaphorase-positive neurons was noted in the superficial dorsal horn layers of lower lumbar and sacral segments. PMID- 10805419 TI - Catalytic nitric oxide synthase activity in the white and gray matter regions of the spinal cord of rabbits. AB - The latest research reveals that nitric oxide as a gas messenger may diffuse into the surrounding extracellular fluid and act locally upon neighboring target cells. However, several observations raise the possibility that nitric oxide may also be released at a greater distance from the neuronal cell body. The catalytic nitric oxide synthase (cNOS) activity was therefore studied in the cervicothoracic and lumbosacral segments of the spinal cord of rabbits, including the white matter of dorsal columns (DC), lateral columns (LC) and ventral columns (VC), as well as the gray matter of dorsal horns (DH), intermediate zone (IZ) and ventral horns (VH). Lower cNOS activity was found in the white matter of both cervicothoracic (47%) and lumbosacral (30%) regions, whereas that detected in the gray matter of the lumbosacral part of the spinal cord was considerably higher (70%). Enzyme activity varied from 43.4 to 77.2 dpm/microg protein in the cervicothoracic segments of the gray matter in the descending order: DH>VH>IZ. Similar cNOS activity was found in the white matter of the cervicothoracic segments (42.1-62.8 dpm/microg protein). When the activity of cNOS was compared in the lumbosacral segments, the highest enzyme activity was found in DH of the gray matter (198.7 dpm/microg protein) and the lowest cNOS in DC (45.8 dpm/microg protein) of the white matter. It was concluded that the white matter of the spinal cord contains similar cNOS activity in comparison to the gray matter. PMID- 10805420 TI - Changes in plasma angiotensin-converting enzyme activity and noradrenaline responses to long-term nitric oxide inhibition vary depending on their basal values in chickens. AB - In this study, we investigated the effects of N(omega)-nitro-L-arginine (L-NNA) on arterial blood pressure (BP), plasma noradrenaline (NA) and adrenaline (A) levels and angiotensin-converting enzyme (ACE) activity. L-NNA was applied with tap water (1 mg/ml) from the 3rd to the 8th week of age (group L-NNA1). In Experiment 1, long-term L-NNA application increased BP compared to the control group (group C1) (L-NNA1 = 131.4 +/- 6.3, n = 6; C1 = 82.7 +/- 4.7 mm Hg, n = 7) but decreased plasma noradrenaline and adrenaline levels and ACE activity (NA levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 8.6 +/- 0.5 ng/ml, n = 7; A levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 6.0 +/- 0.5 ng/ml, n = 7; ACE activities: C1 = 87.3 +/- 3.1, n = 6; L-NNA1 = 46.2 +/- 1.9 U/l, n = 5). On the other hand, in Experiment 2 (carried out under the same conditions and in age-matched chickens), blood pressure, plasma noradrenaline levels and ACE activity were found to differ in the control group (C2) (BP = 141.4 +/- 15.5 mm Hg, n = 7; NA = 1.1 +/- 0.4 ng/ml, n = 7; ACE = 57.2 +/- 5.3 U/l, n = 7) as compared to C1, while plasma adrenaline levels were similar. In this series, long-term L-NNA application (group L-NNA2) did not change the BP, but surprisingly increased noradrenaline and ACE values (values of L-NNA2: BP = 165.7 +/- 15.6 mm Hg, n = 7; NA = 9.3 +/- 1.3 ng/ml, n = 8; ACE = 149.4 +/- 16 U/l, n = 8) while decreasing plasma adrenaline levels. L-arginine addition to L-NNA treatment completely reversed plasma noradrenaline and ACE activity values. These results indicate the modulatory activity of an L-arginine-NO pathway on adrenaline release as well as on the renin-angiotensin system in chickens. PMID- 10805422 TI - Millennium Memorabilia Issue on Apheresis: part 2. Centrifugal devices, adsorbents, leukocyte filters, and extracorporeal immunomodulations. PMID- 10805421 TI - Long-term lisinopril dihydrate application decreases plasma noradrenaline but not adrenaline levels in chickens. AB - Little is known about the effect of chronic angiotensin-converting enzyme inhibition on the catecholamine levels in fowls. In this study, we investigated the effects of chronic lisinopril dihydrate (Ld) application on the plasma levels of adrenaline and noradrenaline and on the blood pressure. Lisinopril was given in different concentrations (25, 75 and 250 mg/l drinking water) to the white Leghorn chickens for 9 weeks, while the control group drank tap water only. Twenty-eight hours after the last lisinopril application, arterial blood pressure (BP), plasma adrenaline and noradrenaline levels, plasma renin (PRA) and plasma angiotensin-converting enzyme (ACE) activities were determined. In all concentrations, lisinopril significantly increased PRA and decreased ACE activities. Arterial BP was decreased only in the group receiving high lisinopril concentration (Controls 119+/-10.27, Ld3 98+/-5.4 mm Hg). However, the lower lisinopril concentrations did not alter arterial BP compared to the control group. Plasma noradrenaline levels were decreased in a concentration-dependent manner (47-58%), but plasma adrenaline levels remained unchanged. The heart weight/body weight ratio was not changed in any of the lisinopril-treated groups. The persistent decrease in the blood pressure after lisinopril treatment was not directly related to a decrease of plasma ACE activity or plasma noradrenaline levels. Its mechanism still remains to be elucidated. PMID- 10805423 TI - New principle of closed system centrifugation. 1956. PMID- 10805424 TI - A simplified centrifuge for the separation and processing of blood cells. 1967. PMID- 10805425 TI - Closed continuous-flow centrifuge. 1968. PMID- 10805426 TI - New flow-through centrifuge without rotating seals applied to plasmapheresis. 1975. PMID- 10805427 TI - Principles of blood separation and component extraction in a disposable continuous-flow single-stage channel. 1979. PMID- 10805428 TI - The development and first clinical use of semipermeable microcapsules (artificial cells) as a compact artificial kidney. 1970. PMID- 10805429 TI - Resin hemoperfusion in the treatment of drug intoxication. 1970. PMID- 10805430 TI - Development of a specific immunoadsorbent containing immobilized synthetic peptide of acetylcholine receptor for treatment of myasthenia gravis. 1993. PMID- 10805432 TI - Development of selective low-density lipoprotein (LDL) apheresis system: immobilized polyanion as LDL-specific adsorption for LDL apheresis system. 1996. PMID- 10805431 TI - Immusorba TR and Immusorba PH: basics of design and features of functions. 1998. PMID- 10805433 TI - A new treatment for endotoxemia with direct hemoperfusion by polymyxin immobilized fiber. 1984. PMID- 10805434 TI - Evaluation of a filtration lymphocytapheresis (LCP) device for use in the treatment of patients with rheumatoid arthritis. 1991. PMID- 10805435 TI - Granulocytapheresis as a possible cancer treatment. 1995. PMID- 10805436 TI - A new device for selective removal of CD4+ T cells. 1998. PMID- 10805437 TI - Efficacy and biocompatibility of a novel anti-cancer fiber in hemoperfusion on cancer-bearing rabbits. 1984. PMID- 10805438 TI - Biocompatibility in membrane plasmapheresis: the necessity of global understanding. 1987. PMID- 10805439 TI - Micromechanics and anatomical changes during early ontogeny of two lianescent Aristolochia species. AB - The mechanical properties of young stems of Aristolochia macrophylla Lam. and Aristolochia brasiliensis Mart. et Zucc. were studied during elongation growth and primary differentiation. Data for the modulus of elasticity, for the viscoelastic behaviour caused by longitudinal tension and for the shear modulus resulting from torsion around a longitudinal axis were related to the underlying structural changes by quantitative analysis of stem anatomy, tissue distribution, ultrastructure, and cell wall biochemistry. The orientation of cellulose microfibrils was determined by light microscopy and small-angle X-ray diffraction, and the lignin content was determined by thioglycolic acid derivatization and spectroscopic quantification. It was demonstrated that the increase in stability during early development is due to the complementary effects of increase in cell wall material, lignification, and cellulose microfibril alignment. A detailed micromechanical model, considering internal prestresses, is proposed to explain the characteristic biphasic stress-strain behaviour as well as the strain-hardening observed. PMID- 10805440 TI - Molecular, functional and ultrastructural characterisation of plastids from six species of the parasitic flowering plant genus Cuscuta. AB - Plastids of Cuscuta reflexa Roxb., C. subinclusa D. et H., C. gronovii Willd. and C. campestris Yunck. possess thylakoids and contain both chlorophyll a and b in a ratio similar to that of stem tissue of the systematically closely related but 'normal' green Ipomoea tricolor. In contrast, plastids of C. odorata R. et P. and C. grandiflora H.B.K. do not contain any chlorophyll or possess thylakoids. Light driven electron transport, as measured by oxygen evolution and indicated by analysis of chlorophyll fluorescence, was present in all chlorophyll-containing species. The photosystem II efficiency was low and ranged from 0.511 to 0.687. The plastid rbcL gene could not be detected in C. odorata, but was present in all other tested species. Neither rbcL transcripts nor the large subunit of ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco) could be detected in C. odorata and C. grandiflora. Low amounts of the large subunit of Rubisco were detected immunologically in all other Cuscuta species. Apparently, the genus Cuscuta comprises species with different degrees of plastid functionality, ranging from intact chloroplasts, via plastids with impaired protein production and gene expression to plastids with reduced plastome gene content. PMID- 10805441 TI - Expression of cucumber lipid-body lipoxygenase in transgenic tobacco: lipid-body lipoxygenase is correctly targeted to seed lipid bodies. AB - A particular isoform of lipoxygenase (LOX, EC 1.13.11.12) localized on lipid bodies has been shown by earlier investigations to play a role during seed germination in initiating the mobilization of triacylglycerols. On lipid bodies of germinating cucumber (Cucumis sativus L.) seedlings, the modification of linoleoyl moieties by this LOX precedes the hydrolysis of the ester bonds. We analyzed the expression and intracellular location of this particular LOX form in leaves and seeds of tobacco (Nicotiana tabacum L.) transformed with one construct coding for cucumber lipid-body LOX and one construct coding for cucumber LOX fused with a hemagglutinin epitope. In both tissues, the amount of lipid-body LOX was clearly detectable. Biochemical analysis revealed that in mature seeds the foreign LOX was targeted to lipid bodies, and the preferred location of the LOX on lipid bodies was verified by immunofluorescence microscopy. Cells of the endosperm and of the embryo exhibited fluorescence based on the immunodecoration of LOX protein whereas very weak fluorescent label was visible in seeds of untransformed control plants. Further cytochemical analysis of transformed plants showed that the LOX protein accumulated in the cytoplasm when green leaves lacking lipid bodies were analyzed. Increased LOX activity was shown in young leaves of transformed plants by an increase in the amounts of endogenous (2E) hexenal and jasmonic acid. PMID- 10805442 TI - Identification and expression of three new Nicotiana plumbaginifolia genes which encode isoforms of a plasma-membrane H(+)-ATPase, and one of which is induced by mechanical stress. AB - To analyze in detail the multigene family encoding the plasma-membrane H(+) ATPase (pma) in Nicotiana plumbaginifolia Viv., five new pma genes (pma 5-9) were isolated. Three of these (pma 6, 8, 9) were fully characterized and classified into new and independent subfamilies. Their cell-type expression was followed by the beta-glucuronidase (gusA) reporter-gene method. While the pma8-gusA transgene was not expressed in transgenic tobacco, expression of the two other transgenes (pma6- and pma9-gusA) was found to be restricted to particular cell types. In the vegetative tissues, pma6-gusA expression was limited to the head cells of the leaf short trichomes, involved in secretion, and to the cortical parenchyma of the young nodes where the developing leaves and axillary flowering stalks join the stem. In the latter tissues, gene expression was enhanced by mechanical stress, suggesting that H(+)-ATPase might be involved in the strength of the tissues and their resistance to mechanical trauma. The pma9-gusA transgene was mainly expressed in the apical meristem of adventitious roots and axillary buds as well as in the phloem tissues of the stem, in which expression depended on the developmental stage. In flowers, pma9-gusA expression was limited to the mature pollen grains and the young fertilized ovules, while that of pma6-gusA was identified in most of the organs. Reverse transcription-polymerase chain reaction of leaf and stem RNA confirmed the expression of pma 6 and 9, while pma8 was found to be expressed in both organs at a lower level. In conclusion, although pma 6 and 9 had a more restricted expression pattern than the previously characterized pma genes, they were nevertheless expressed in cell types in which H(+)-ATPase had not been previously detected. PMID- 10805443 TI - Functional characterisation of LKT1, a K+ uptake channel from tomato root hairs, and comparison with the closely related potato inwardly rectifying K+ channel SKT1 after expression in Xenopus oocytes. AB - A cDNA encoding a novel inwardly rectifying potassium (K+(in)) channel, LKT1, was cloned from a root-hair-specific cDNA library of tomato (Lycopersicon esculentum Mill.). The LKT1 mRNA was shown to be most strongly expressed in root hairs by Northern blot analysis. The LKT1 channel is a member of the AKT family of K+(in) channels previously identified in Arabidopsis thaliana (L.) Heynh. and potato (Solanum tuberosum L.). Moreover, LKT1 is closely related (97% identical amino acids) to potato SKT1. An electrophysiological comparison of the two channels should therefore assist the identification of possible molecular bases for functional differences. For, this comparison, both channels were functionally expressed and electrophysiologically characterised within the same expression system, i.e. Xenopus laevis oocytes. Voltage-clamp measurements identified LKT1 as a K(+)-selective inward rectifier which activates with slow kinetics upon hyperpolarising voltage pulses to potentials more negative than -50 mV. The activation potential of LKT1 is shifted towards positive potentials with respect to SKT1 which might be due to single amino acid exchanges in the rim of the channel's pore region or in the S4 domain. Like SKT1, LKT1 reversibly activated upon shifting the external pH from 6.6 to 5.5, which indicates a physiological role for pH-dependent regulation of AKT-type K+(in) channels. The pharmacological inhibitor Cs+, applied externally, inhibited K+(in) currents mediated by LKT1 and SKT1 half-maximally with a concentration (IC50) of 21 microM and 17 microM, respectively. In conclusion, LKT1 may serve as a low-affinity influx pathway for K+ into root hair cells. Comparison of homologous K+(in) rectifiers from different plant species expressed in the same heterologous system allows conclusions to be drawn in respect to structure-function relationships. PMID- 10805444 TI - Differential localization of arabinan and galactan side chains of rhamnogalacturonan 1 in cambial derivatives. AB - The development of pectin structural features during the differentiation of cambial derivatives was investigated in aspen (Populus tremula L. x P. tremuloides Michx.) using biochemical and immunocytochemical methods. Comparisons were also made between active and resting tissues. Active tissues, in particular cambial cells and phloem derivatives, were characterized by a high pectin content. Use of antibodies raised against arabinan side chains of rhamnogalacturonan 1 (LM6), as well as biochemical analysis, revealed an obvious decrease from the cortex to the differentiating xylem. Galactan side chains, detected with LM5 antibodies, were present mainly in the cambial zone and enlarging xylem cells. In contrast, they were totally absent from sieve-tube cell walls. Image analysis of LM5 immunogold labelling in the cambial zone showed a clustered distribution of galactan epitopes in the radial walls, a distribution which might result from the association of two different periodic processes, namely the exocytosis of galactan and wall expansion. Cessation of cambial activity was characterized by cell wall thickening accompanied by a sharp decrease in the relative amount of pectin and a lowering of the degree of methylesterification. The data provide evidence that the walls of phloem and xylem cells differ in their pectin composition even at a very early stage of commitment. These differences offer useful tools for identifying the initial cells among their immediate neighbours. PMID- 10805445 TI - Water-selective and multifunctional aquaporins from Lotus japonicus nodules. AB - By using reverse transcriptase-polymerase chain reaction, two cDNAs were isolated that encode major intrinsic membrane proteins (MIPs) that are expressed in nitrogen-fixing root nodules of Lotus japonicus. Lotus intrinsic membrane protein 1 (LIMP 1) is expressed at high levels in both nodule and root tissues and shows highest sequence similarity to members of the tonoplast intrinsic protein (TIP) subfamily of plant MIPs. Functional analysis of LIMP 1 by expression in Xenopus laevis oocytes show that it is a water-specific aquaporin. In contrast, LIMP 2 shows the highest sequence similarity to soybean nodulin 26 (67.8% amino acid sequence identity). LIMP 2 is also a nodulin, showing expression only in mature nitrogen fixing nodules of L. japonicus. LIMP 2 is a multifunctional aquaglyceroporin, and displays the ability to flux both water as well as glycerol upon expression in Xenopus oocytes. Additionally, the carboxyl terminal region of LIMP 2 has a conserved phosphorylation motif that is phosphorylated by a calmodulin-like domain protein kinase. Overall, the data show that L. japonicus nodules contain two structurally and functionally distinct MIP proteins: one (LIMP 2) which appears to be the nodulin 26 ortholog of L. japonicus and another (LIMP 1) which appears to be a member of the TIP subfamily. PMID- 10805446 TI - Late steps of egg cell differentiation are accelerated by pollination in Zea mays L. AB - Egg cells were analysed cytologically during the female receptivity period in maize (Zea mays L., line A 188). Three classes of egg cell were distinguished: type A--small, non-vacuolated cells with a central nucleus; type B--larger cells with small vacuoles surrounding the perinuclear cytoplasm located in the middle of the cell; type C--big cells with a large apical vacuole and the mid-basal perinuclear cytoplasm. The less-dense cytoplasm of the vacuolated egg cells usually contained numerous cup- or bell-shaped mitochondria. The three egg types appear to correspond to three late stages of egg cell differentiation. The frequencies of each of the three egg types were monitored in developing maize ears before and after pollination. In young ears, with the silks just extending out of the husks, small A-type cells were found in about 86% of ovules. Their frequency decreased to about 58% at the optimum silk length, remained unchanged in non-pollinated ears, and fell to 16% at the end of the female receptivity period. However, after pollination and before fertilisation the frequency of these cells decreased to about 33%, and the larger vacuolated egg cells (types B and C) prevailed. At various stages of the receptivity period, pollination accelerated changes in the egg population, increasing the number of ovules bearing larger, vacuolated egg cells. Experiments with silk removal demonstrated that putative pollination signals act immediately after pollen deposition and are not species-specific. PMID- 10805447 TI - Affinity for inorganic carbon of Gracilaria tenuistipitata cultured at low and high irradiance. AB - Regulation by irradiance level of the mechanism for dissolved inorganic carbon (DIC) acquisition was examined in the red macroalga Gracilaria tenuistipitata Zhang et Xia. For this purpose, affinity for external DIC, carbonic anhydrase (CA; EC 4.2.1.1) activity and content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) were determined in thalli grown at 45 and 500 micromol photons m(-2) s(-1). Oxygen evolution rates declined by 50% when the medium pH was changed from 8.1 to 8.7, and the pH compensation point attained was ca. 9.2. These characteristics were unaffected by the light treatments. In contrast, photosynthetic conductance for DIC at pH 8.7 was doubled in thalli grown at high irradiance compared with those grown at low irradiance (to 0.74 x 10(-6) from 0.33 x 10(-6) m s(-1)). Photosynthetic rates at saturating DIC concentration were also higher by 60% in thalli grown at high irradiance. These differences could not be attributed to changes in the use of external DIC, since external CA activity did not vary. Although the irradiance level did not modify the pool size of Rubisco, Rubisco content expressed on a chlorophyll a basis was almost doubled at high irradiance. These results likely indicate that the internal transport of DIC towards the active-site of Rubisco, rather than the external use of DIC, is enhanced in the thalli grown at high irradiance. PMID- 10805448 TI - Influence of ascorbate and the Mehler peroxidase reaction on non-photochemical quenching of chlorophyll fluorescence in maize mesophyll chloroplasts. AB - Non-photochemical quenching of chlorophyll fluorescence (NPQ) and quantum yield of photosystem II (PSII) were studied with intact mesophyll chloroplasts of maize (Zea mays L.) during the initial minutes of illumination using the pulse modulated chlorophyll fluorescence technique. Non-photochemical quenching was rapidly reversible in the dark at any point during illumination, which is indicative of energy-dependent dissipation of energy (mediated via thylakoid delta pH changes and ascorbate-dependent synthesis of zeaxanthin). In chloroplasts suspensions including 15 mM ascorbate in the medium, with addition of oxaloacetate and pyruvate, the PSII yield, rate of reduction of oxaloacetate and phosphorylation of pyruvate reached a maximum after approximately 2 min of illumination. Under these conditions, which promote phosphorylation and a decreased delta pH across the thylakoid membrane, NPQ rose to a maximum after 2-3 min of illumination, dropped to a minimum after about 6 min, and then increased to a steady-state level. A rather similar pattern was observed when leaves were illuminated following a 30-min dark period. Providing chloroplasts with higher levels of ascorbate (60 mM), prevented the transient drop in NPQ. Anaerobic conditions or addition of potassium cyanide caused a decrease in PSII yield, providing evidence for operation of the ascorbate-dependent Mehler-peroxidase reaction. These conditions also strongly suppressed the transient drop in NPQ. Dithiothreitol, an inhibitor of violaxanthin de-epoxidase, caused a large drop in NPQ even in the presence of high levels of ascorbate. The results suggest that the decline of NPQ occurs in response to an increase in lumen pH after initiation of phosphorylation, that this decline can be suppressed by conditions where ascorbate is not limiting for violaxanthin de-epoxidase, and that the increase of NPQ after such a decline is the result of development of energy dissipation in PSII reaction centers. PMID- 10805449 TI - Ferric reduction by iron-limited Chlamydomonas cells interacts with both photosynthesis and respiration. AB - Iron limitation led to a large increase in extracellular ferricyanide (Fe[III]) reductase activity in cells of the green alga Chlamydomonas reinhardtii Dangeard. Mass-spectrometric measurement of gas exchange indicated that ferricyanide reduction in the dark resulted in a stimulation of respiratory CO2 production without affecting the rate of respiratory O2 consumption, consistent with the previously postulated activation of the oxidative pentose phosphate pathway in support of Fe(III) reduction by iron-limited Chlamydomonas cells (X. Xue et al., 1998, J. Phycol. 34: 939-944). At saturating irradiance, the rate of ferricyanide reduction was stimulated almost 3-fold, and this stimulation was inhibited by 3 (3',4'-dichlorophenyl)-1,1-dimethylurea. Ferricyanide reduction during photosynthesis resulted in approximately a 50% inhibition of photosynthetic CO2 fixation at saturating irradiance, and almost 100% inhibition of CO2 fixation at sub-saturating irradiance. Photosynthesis by iron-sufficient cells was not affected by ferricyanide addition. Addition of 250 microM ferricyanide to iron limited cells in which photosynthesis was inhibited (either by the presence of glycolaldehyde, or by maintaining the cells at the CO2 compensation point) resulted in a stimulation in the rate of gross photosynthetic O2 evolution. Chlorophyll a fluorescence measurements indicated a large increase in non photochemical quenching during ferricyanide reduction in the light; the increase in nonphotochemical quenching was abolished by the addition of nigericin. These results suggest that reduction of extracellular ferricyanide (mediated at the plasma membrane) interacts with both photosynthesis and respiration, and that both of these processes contribute NADPH in the light. PMID- 10805451 TI - Different pH-dependences of K+ channel activity in bundle sheath and mesophyll cells of maize leaves. AB - The isolation of bundle sheath protoplasts from leaves of Zea mays L. for patch clamp whole-cell experiments presents special problems caused by the suberin layer surrounding these cells. These problems were overcome by the isolation technique described here. Two different types of whole-cell response were found: a small response caused by MB-1 (maize bundle sheath conductance type 1) which was instantaneously activated, and another caused by MB-2 (maize bundle sheath conductance type 2) consisting of an instantaneous response (maize bundle sheath K+ instantaneous current type 2; MB-KI2) similar to but stronger than the current through MB-1 plus a small time-dependent outward rectifying component (maize bundle sheath activated outward rectifying current; MB-AOR) with voltage dependent delayed activation. The occurrence of MB-AOR was often accompanied by a smaller contribution from an inward rectifying channel at negative potentials. Activation of MB-2 required ATP. It is suggested that MB-1 and MB-2 are related to bundle sheath cells with and without direct contact with the xylem vessels. In mesophyll cells, only one type of response caused by MM-2 (maize mesophyll conductance type 2) was found with an instantaneous (maize mesophyll K+ instantaneous current type 2, MM-KI2) and a voltage-dependent delayed component (maize mesophyll activated outward rectifying current, MM-AOR). The most striking difference between bundle sheath and mesophyll cells was the pH dependence of K+ uptake. At pH 7.2, uptake of K+ by MB-2 was identical to that by MM-2 over the whole voltage range. However, acidification stimulated K+ conductance in bundle sheath cells, whereas a decrease was found for MM-2. At pH 6.15, the bundle sheath channel MB-2 had more than a 10-fold higher K+ uptake at positive and negative potentials than MM-2. The channel MB-1, too, was stimulated by low pH. This seems to indicate a putative role for MB-1 and MB-2 in charge balance during uptake of nutrients via cotransport from the xylem into the symplasm. PMID- 10805450 TI - Characterization of pectin methyltransferase from soybean hypocotyls. AB - Pectin methyltransferase (PMT) catalyzing the transfer of the methyl group from S adenosyl-L-methionine (SAM) to the C-6 carboxyl group of galactosyluronic acid residues in pectin was found in a membrane preparation of etiolated hypocotyls from 6-d-old soybean (Glycine max Merr.). The enzyme was maximally active at pH 6.8 and 35-40 degrees C, and required 0.5% (w/v) Triton X-100. The incorporation of the methyl group was significantly enhanced by addition of a pectin with a low (22%) degree of methyl-esterification (DE) as exogenous acceptor substrate. The apparent Michaelis constants for SAM and the pectin (DE22) were 0.23 mM and 66 microg x ml(-1), respectively. Attachment of the methyl group to the carboxyl group of the pectin via ester linkage was confirmed by analyzing radiolabeled product from incubation of the enzyme with [14C]methyl SAM and the acceptor pectin. Size-exclusion chromatography showed that both enzymatic hydrolysis with a pectin methylesterase and a mild alkali treatment (saponification) led to the release of radioactive methanol from the product. Enzymatic hydrolysis of the product with an endopolygalacturonase degraded it into small pectic fragments with low relative molecular mass, which also supports the idea that the methyl group is incorporated into the pectin. The soybean hypocotyls were fractionated into their cell wall components by successive extraction with water, EDTA, and alkali treatment. Among the resulting polysaccharide fractions, high PMT activity was observed when a de-esterified polysaccharide derived from the EDTA-soluble fraction (the pectic fraction) was added as an alternative acceptor substrate, indicating that the enzyme may be responsible for producing methyl-esterified pectin in vivo. PMID- 10805452 TI - Discrepancy between nitrate reduction rates in intact leaves and nitrate reductase activity in leaf extracts: what limits nitrate reduction in situ? AB - Nitrate reductase (NR) activity in spinach leaf extracts prepared in the presence of a protein phosphatase inhibitor (50 microM cantharidine) was measured in the presence of Mg2+ (NRact) or EDTA (NRmax), under substrate saturation. These in vitro activities were compared with nitrate reduction rates in leaves from nitrate-sufficient plants. Spinach leaves containing up to 60 micromol nitrate per g fresh weight were illuminated in air with their petiole in water. Their nitrate content decreased with time, permitting an estimation of nitrate reduction in situ. The initial rates (1-2 h) of nitrate consumption were usually lower than NRact, and with longer illumination time (4 h) the discrepancy grew even larger. When leaves were fed through their petiole with 30 mM nitrate, initial in-situ reduction rates calculated from nitrate uptake and consumption were still lower than NRact. However, nitrate feeding through the petiole maintained the in situ-nitrate reduction rate for a longer time. Initial rates of nitrate reduction in situ only matched NRact when leaves were illuminated in 5% CO2. In CO2-free air or in the dark, both NRact and in-situ nitrate reduction decreased, but NRact still exceeded in-situ reduction. More extremely, under anoxia or after feeding 5-amino-4-imidazole carboxyamide ribonucleoside in the dark, NR was activated to the high light level; yet in spite of that, nitrate reduction in the leaf remained very low. It was examined whether the standard assay for NRact would overestimate the in-situ rates due to a dissociation of the inactive phospho-NR-14-3-3 complex after extraction and dilution, but no evidence for that was found. In-situ NR obviously operates below substrate saturation, except in the light at high ambient CO2. It is suggested that in the short term (2 h), nitrate reduction in situ is mainly limited by cytosolic NADH, and cytosolic nitrate becomes limiting only after the vacuolar nitrate pool has been partially emptied. PMID- 10805453 TI - Drought induces fructan synthesis and 1-SST (sucrose:sucrose fructosyltransferase) in roots and leaves of chicory seedlings (Cichorium intybus L.). AB - Seeds of Cichorium intybus L. var. foliosum cv. Flash were sown in acid-washed vermiculite and grown in a controlled-environment growth chamber. After 1 month of growth, plantlets did not contain sucrose:sucrose 1-fructosyltransferase (1 SST), the key enzyme in fructan biosynthesis. No fructan could be observed. Some of the plants were submitted to drought for 2 weeks. Glucose, fructose and sucrose concentrations increased in roots and leaves of stressed plants and the fructan concentration in roots and leaves was ten times higher than in control plants. The onset of fructan synthesis coincided with the increase in 1-SST activity in roots. Expression of the 1-SST gene could be observed in roots and leaves of stressed plants. PMID- 10805454 TI - Demonstration and characterization of (E)-nerolidol synthase from maize: a herbivore-inducible terpene synthase participating in (3E)-4,8-dimethyl-1,3,7 nonatriene biosynthesis. AB - Upon herbivore attack, maize (Zea mays L.) emits a mixture of volatile compounds that attracts herbivore enemies to the plant. One of the major components of this mixture is an unusual acyclic C11 homoterpene, (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), which is also emitted by many other species following herbivore damage. Biosynthesis of DMNT has been previously shown to proceed via the sesquiterpene alcohol, (E)-nerolidol. Here we demonstrate an enzyme activity that converts farnesyl diphosphate, the universal precursor of sesquiterpenes, to (3S)-(E) nerolidol in cell-free extracts of maize leaves that had been fed upon by Spodoptera littoralis. The properties of this (E)-nerolidol synthase resemble those of other terpene synthases. Evidence for its participation in DMNT biosynthesis includes the direct incorporation of deuterium-labeled (E)-nerolidol into DMNT and the close correlation between increases in (E)-nerolidol synthase activity and DMNT emission after herbivore damage. Since farnesyl diphosphate has many other metabolic fates, (E)-nerolidol synthase may represent the first committed step of DMNT biosynthesis in maize. However, the formation of this unusual acyclic terpenoid appears to be regulated at both the level of (E) nerolidol synthase and at later steps in the pathway. PMID- 10805455 TI - Isolation and characterisation of cell wall polysaccharides from cocoa (Theobroma cacao L.) beans. AB - Cell wall material (CWM) was prepared from sun-dried cocoa (Theobroma cacao L.) bean cotyledons before and after fermentation. The monosaccharide composition of the CWM was identical for unfermented and fermented beans. Polysaccharides of the CWM were solubilised by sequential extraction with 0.05 M trans-1,2 diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA), 0.05 M Na2CO3, and 1 M, 4 M and 8 M KOH. The non-cellulosic sugar composition for each fraction was similar for unfermented and fermented samples, indicating that fermentation caused no significant modification of the structural features of individual cell wall polysaccharides. Pectic polysaccharides accounted for 60% of the cell wall polysaccharides but only small amounts could be solubilised in solutions of CDTA, Na2CO3, and 1 M and 4 M KOH. The bulk of the pectic polysaccharides were solubilised in 8 M KOH and were characterised by a rhamnogalacturonan backbone heavily substituted with side-chains of 5-linked arabinose and 4-linked galactose. Linkage analysis indicated the presence of additional acidic polysaccharides, including a xylogalacturonan and a glucuronoxylan. Cellulose, xyloglucan and a galactoglucomannan accounted for 28%, 8% and 3% of the cell wall polysaccharides, respectively. It is concluded that the types and structural features of cell wall polysaccharides in cocoa beans resemble those found in the parenchymatous tissue of many fruits and vegetables rather than those reported for many seed storage polysaccharides. PMID- 10805456 TI - Conversion of guaiacyl to syringyl moieties on the cinnamyl alcohol pathway during the biosynthesis of lignin in angiosperms. AB - Aglycons derived from 4-O-beta-D-glucosides of both caffeyl and 5 hydroxyconiferyl alcohols were incorporated into guaiacyl (G) and syringyl (S) units in the lignin of newly formed xylem of several angiosperms. It is likely that these aglycons enter the cinnamyl alcohol pathway as intermediates in the introduction of methoxyl groups onto aromatic rings, and serve as precursors for the biosynthesis of lignin. The S/G ratio in this pathway was coincident with the ratio in the cell wall lignin of each tree. Our results indicate that the cinnamyl alcohol pathway involves the same mechanisms as the cinnamic acid and cinnamyl CoA pathways and they suggest that this novel pathway might be part of a metabolic grid in the biosynthesis of lignin. PMID- 10805457 TI - The role of plant villin in the organization of the actin cytoskeleton, cytoplasmic streaming and the architecture of the transvacuolar strand in root hair cells of Hydrocharis. AB - In many types of plant cell, bundles of actin filaments (AFs) are generally involved in cytoplasmic streaming and the organization of transvacuolar strands. Actin cross-linking proteins are believed to arrange AFs into the bundles. In root hair cells of Hydrocharis dubia (Blume) Baker, a 135-kDa polypeptide cross reacted with an antiserum against a 135-kDa actin-bundling protein (135-ABP), a villin homologue, isolated from lily pollen tubes. Immunofluorescence microscopy revealed that the 135-kDa polypeptide co-localized with AF bundles in the transvacuolar strand and in the sub-cortical region of the cells. Microinjection of antiserum against 135-ABP into living root hair cells induced the disappearance of the transvacuolar strand. Concomitantly, thick AF bundles in the transvacuolar strand dispersed into thin bundles. In the root hair cells, AFs showed uniform polarity in the bundles, which is consistent with the in-vitro activity of 135-ABP. These results suggest that villin is a factor responsible for bundling AFs in root hair cells as well as in pollen tubes, and that it plays a key role in determining the direction of cytoplasmic streaming in these cells. PMID- 10805458 TI - Identification of a signal peptide for oryzacystatin-I. AB - A previously unidentified extension of an open reading frame from the genomic DNA of Japonica rice (Oryza sativa L.) encoding oryzacystatin-I (OC-I; access. M29259, protein ID AAA33912.1) has been identified as a 5' gene segment coding for the OC-I signal peptide. The signal peptide appears to direct a pre-protein (SPOC-I; Accession No. AF164378) to the endoplasmic reticulum, where it is processed into the mature form of OC-I. The start codon of SPOC-I begins 114 bp upstream from that previously published for OC-I. A putative proteolytic site. which may yield a mature OC-I approximately 12 residues larger than previously described, has been identified within SPOC-I between Ala-26 and Glu-27. The signal peptide sequence was amplified by polymerase chain reaction using genomic DNA from O. sativa seedlings and ligated to the 5' end of the truncated OC-I gene at the endogenous SalI site. Partially purified protein extracts from Escherichia coli expressing SPOC-I reacted with polyclonal antibodies raised against OC-I and revealed a protein of the expected molecular weight (15,355 Da). In-vitro translation of SPOC-I in the presence of microsomal membranes yielded a processed product approximately 2.7 kDa smaller than the pre-protein. Nicotiana tabacum L. cv. Xanthi plants independently transformed with the SPOC-I gene processed SPOC-I and accumulated the mature form of OC-I (approximately 12.6 kDa), which co migrated with natural, mature OC-I extracted from rice seed when separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 10805459 TI - Gravity-induced absorbance changes in Phycomyces: a novel method for detecting primary responses of gravitropism. AB - The negative gravitropism of the sporangiophores of Phycomyeces blakesleeanus Burgeff is elicited by different sensory inputs, which include flexure of the growing zone, buoyance of lipid globules and sedimentation of paracrystalline proteins, so-called octahedral crystals (C. Schimek et al., 1999a, Planta 210: 132-142). Gravity-induced absorbance changes (GIACs), which are associated with primary events of gravity sensing, were detected in the growing zones of sporangiophores. After placing sporangiophores horizontally, GIACs were detected after a latency of about 5 min, i.e. 15-25 min prior to gravitropic bending. The spectroscopic properties of the GIACs indicate that gravitropic stimulation could imply the reduction of cytochromes. The GIACs were spectrally distinct from light induced absorbance changes (LIACs), showing that the primary responses of the light and gravity transduction chains are different. A dual stimulation with gravity and light generated GIAC-LIACs which were distinct from the absorbance changes occurring after the single stimuli and which indicate that light and gravity interact early in the respective transduction chains. PMID- 10805460 TI - Recurrent focal segmental glomerulosclerosis in grafts treated with plasma exchange and increased immunosuppression. AB - We report on three children with severe, recurrent focal segmental glomeruloscerosis (FSGS) in their first allografts, treated with methylprednisolone, plasma exchange and cyclophosphamide. This protocol is based on a previous publication showing its successful use in three children. Our patients were 2 girls and 1 boy, aged 14.5, 14.6 and 13.2 years, respectively, at transplant. Concomitant immunosuppression included cyclosporin A and prednisolone. Recurrence occurred in all three patients within 24 h, and specific treatment was commenced within 48 h. All patients developed anuria and were dialysed. The boy stopped dialysis after 4 weeks, and has stable chronic renal failure (CRF) and no proteinuria 3 years later. One girl required dialysis for 4 months, and 3 years later has CRF with non-nephrotic range proteinuria. The other girl remained dialysis-dependent and died from septic complications. We conclude that even anuric patients treated with this protocol may have an improvement in renal function and reduction of proteinuria, which can last for over 3 years. However, treatment may need to be prolonged and carries the substantial risks of heavy immunosuppression. PMID- 10805461 TI - Paecilomyces variotii peritonitis in an infant on automated peritoneal dialysis. AB - Fungal peritonitis is a serious complication of chronic peritoneal dialysis (CPD) and is frequently associated with CPD drop-out. Paecilomyces variotii, a common saprophytic fungus, rarely causes human infection. To date, only nine adult or adolescent patients with P. variotii peritonitis during continuous ambulatory peritoneal dialysis have been reported. In all patients, successful treatment required antifungal therapy and removal of the peritoneal catheter. We report the first case of P. variotii peritonitis in an infant on automated peritoneal dialysis successfully treated with combined intraperitoneal and oral fluconazole, without removal of the peritoneal catheter. PMID- 10805462 TI - Potential prophylactic use of benzodiazepines for hemodialysis-associated seizures. AB - Hemodialysis-associated seizure (HAS) is a common complication of hemodialysis. The efficiency of anticonvulsant drugs in treating or preventing seizures is poorly defined. In this study, children on long-term hemodialysis were examined for HAS and the effect of diazepam prophylaxis on HAS was investigated. Nine patients with a mean age of 14.1 + 2.8 years had HAS and 4 with a mean age of 13.0 +/- 4.4 years had never experienced HAS. The patients with HAS had tonic clonic seizures. Four patients had focal slow-wave paroxysms, especially in the parieto-occipital regions; 2 had subcortical epileptiform discharges by electroencephalography. No correlation was observed between HAS and patient age, primary disease, prior history of seizures, type of dialysis, duration of hemodialysis, anemia, hyperparathyroidism, and administration of erythropoietin. Hypertension due to hypervolemia may also play a role in the development of HAS. Five patients with HAS first treated with phenobarbital (PB) had recurrence of seizures. As a dialyzable antiepileptic PB may be associated with an increased risk for HAS. In a preliminary study, we gave diazepam as a prophylactic therapy to 4 patients with HAS. During 6 months of follow-up, these patients had no seizures. The number of HAS was significantly different between the groups receiving PB and diazepam (z=-2,58, P=0.009). In conclusion, administration of diazepam per os to patients with HAS may be of value for preventing recurrence of HAS. PMID- 10805463 TI - Acute cyclosporine A-induced nephrotoxicity: a rabbit model. AB - Chronic cyclosporine A (CsA) nephrotoxicity has been widely assessed but only few studies have described acute nephrotoxicity. As CsA is now used for short periods, we developed an experimental model of acute CsA-induced nephrotoxicity. Renal clearances of inulin and para-aminohippurate were assessed in 35 New Zealand rabbits. Group 1: control, no treatment; group 2: CsA 25 mg/kg per day in 0.5 ml/kg per day for 5 days; group 3: vehicle Cremophor-EL, 0.5 ml/kg per day for 5 days; group 4: follow-up, the same as group 2, then CsA discontinuation for 31 days. Compared with group 1, CsA significantly decreased glomerular filtration rate (GFR), renal blood flow (RBF), and diuresis, with a significant increase in renal vascular resistance (RVR). The proportional fall in GFR (-32.3%) and RBF ( 33.1%) suggests both pre- and postglomerular vasoconstriction. Discontinuation of CsA in group 4 led to normalization of RVR with improvement of other renal function parameters. Compared with group 1, Cremophor-EL induced no significant changes but an increased RBF. Microvacuolization of proximal tubule epithelial cells was the sole histological abnormality observed only in group 2. The overall results suggest that CsA induced a vasomotor acute renal failure which was not due to Cremophor-EL. This effect was partly reversible after discontinuation of treatment. PMID- 10805464 TI - Renal polyamine excretion, tubular amino acid reabsorption and molecular genetics in cystinuria. AB - Cystinuria is an autosomal recessive disorder of the tubular and intestinal resorption of cystine, ornithine. lysine and arginine leading to nephrolithiasis. Three cystinuria types can be distinguished by the mode of inheritance (true recessive or intermediate) and by the pattern of the intestinal amino acid transport. In the present study phenotypes were assessed by the urinary excretion of amino acids related to creatinine, the percentage tubular amino acid reabsorption and the urinary excretion of polyamines as a possible indicator of the intestinal transport defect. However, our thorough phenotyping did not reveal more than two cystinuria types. Genotypes were examined in linkage analyses and single-strand conformation polymorphism-based mutation identification. The SLC3A1 mutations M467T and T216M were disease causing in our homozygous patients of type I cystinuria. We can show the association of type I cystinuria with SLC3A1 and of non-type I cystinuria with a yet unidentified gene on chromosome 19q13.1. Our phenotype and genotype analyses provide evidence for only two types of cystinuria in the investigated patient cohort. PMID- 10805465 TI - The etiology of renal scars in infants with pyelonephritis and vesicoureteral reflux. AB - We aimed to investigate, by means of dimercaptosuccinic acid (DMSA) scan, the relations between vesicoureteral reflux (VUR) and its degree, pyelonephritis during infancy, and renal parenchymal findings. Seventy-four infants with pyelonephritis, 44 girls and 30 boys (mean age at their first pyelonephritic episode 4.12 months, median 3 months), were enrolled in the study. Voiding cystourethrography (VCU) and ultrasonography (US) were performed within 6 weeks following the infection. DMSA was performed at least 4 months after the urinary tract infection (UTI). The renal parenchymal pathology was defined as focal or multifocal defects or as a split renal uptake of less than 45%. DMSA scintigraphy revealed that 19% (14/74) of the children had renal damage. Renal parenchymal findings were observed only when VUR was present, and its grade was above 3/5. No abnormality was found in 51 renal units without reflux, 9 with VUR grade 1/5, and 54 with grade 2/5. Renal pathology was observed in 9/24 renal units with VUR grade 3, 3/8 with grade 4, and 2/2 with grade 5. No correlation was found between renal parenchymal defects and clinical presentation of the pyelonephritis, type of the microorganism, presence of bacteremia, or the number of recurrent infections. In adequately treated infants, renal damage is probably due to a reflux-associated, preexisting, congenital renal parenchymal pathology and not to the inflammatory process. We suggest that DMSA scintigraphy should not be performed routinely in every infant with UTI and should be reserved primarily for children with VUR grade 3 and above. PMID- 10805466 TI - Expression of Ki-67 antigen using monoclonal antibody MIB-1 in children with post streptococcal glomerulonephritis. AB - We investigated the expression of Ki-67 antigen using monoclonal antibody MIB-1 in glomeruli and renal tubules of 21 children (18 males, 3 females) with post streptococcal glomerulonephritis (PSGN). Patients were divided into two groups of active and convalescent phases. The active group (n=13) comprised those patients with clinical manifestations of the acute nephritic syndrome consisting of edema, hypertension, hematuria, and oliguria or those in whom percutaneous renal biopsy was performed within 4 weeks of onset of the symptoms of PSGN and those with serum C3 levels below 55 mg/dl at the time of biopsy. MIB-1 expression was considered positive when staining of endocapillary cells was observed. Of the 21 biopsies, expression of MIB-1 in glomeruli and renal tubules was observed in 14 cases (63.6%) and 20 cases (95.7%), respectively. The expression of MIB-1 in glomeruli of patients with active disease (11/13, 84.6%) was significantly higher than that of the convalescent group (2/8, 25%) (P=0.018). The cellularity in the glomeruli was more severe in the active group than the convalescent group (P=0.0475). There was a significant difference of neutrophilic infiltration in glomeruli between the active group and the convalescent group (P=0.0117). However, glomerular MIB-1 expression did not correlate with the degree of immunofluorescence, the number of neutrophils in the glomeruli on light microscopy, and the presence of subepithelial dense deposits on electron microscopy. There was no significant correlation between MIB-1 and serum C3 level. There was no significant correlation between glomerular MIB-1 expression and creatinine clearance (r=-0. 180, P=0.556) or 24-h urinary protein excretion (r=0.434, P=0.137). Our results suggest that the expression of MIB- in glomeruli in the active phase in PSGN was higher than in the convalescent phase and expression of glomerular MIB-1 appears to be related to glomerular endocapillary proliferation with exudative lesions in children with PSGN. PMID- 10805467 TI - Fetal renal artery flow and renal echogenicity in the chronically hypoxic state. AB - The object of this study was to investigate the fetal renal arterial blood flow in normal and hyperechogenic kidneys during the third trimester of gestation. The pregnancies screened were all chronically hypoxic. Depending on the etiology of the intrauterine chronic hypoxia, the cases were divided into two study groups. Group I comprised 120 pregnant women with pregnancy-associated hypertension and/or proteinuria. Group II consisted of 87 pregnancies with intrauterine growth retardation. Both study groups included pregnant women from the third trimester. Hyperechogenic renal medullae were detected in 15 out of 120 cases with pregnancy associated hypertension and/or proteinuria, and in 22 fetuses of the 87 pregnancies involving intrauterine growth retardation. Fetal renal hyperechogenicity appears to be an indicator of fetal arterial circulatory depression, correlated with pathological changes in the resistance index for the fetal renal arteries. The fetal renal arterial blood flow resistance index was significantly lower in hyperechogenic cases. This may also be an in utero indication of subsequent intrauterine and neonatal complications, such as cesarean section because of fetal distress (43%), treatment in a neonatal intensive care unit (51%) or increased perinatal mortality (5.4%, as compared with 0.8-1.0% in the normal population). Detailed ultrasound and Doppler examinations of renal parenchyma and arteries appear to be useful methods in the prenatal diagnosis of reduced renal perfusion and of intrauterine hypoxia to detect possible pathological fetal conditions in utero. PMID- 10805468 TI - Membranoproliferative glomerulonephritis in two siblings: report and literature review. AB - There is evidence of a genetic basis in some cases of idiopathic membranoproliferative glomerulonephritis (MPGN) types I and III, particularly those occurring in families. The clinical and morphological features and disease course in two siblings with MPGN are described. In the male sibling, both clinical and morphological features as well as serum complement profile suggested type I MPGN; electron microscopy appearance in the female sibling was consistent with type III MPGN. Both patients had treatment-resistant nephrotic syndrome which evolved into renal insufficiency in the girl. No hereditary complement deficiencies were found in siblings or their parents. Both children exhibited HLA A24; -B27, w4; -DR11, 52; -DQ3 antigens. Between 1981 and 1996, 18 patients from eight families with unequivocal diagnosis of MPGN I or III had been described. The mode of inheritance appeared to be autosomal dominant or X-linked in four of these families. In 11 patients, including our 2, in whom HLA typing was performed, eight had the HLA-A2 antigen. Similarities and discrepancies regarding clinical and morphological features and outcomes were evident in these intrafamilial cases, suggesting either a similar genetic background or a multigenic origin of MPGN. The familial occurrence of the MPGN, highlighted by our report, supports the concept that genetically determined factors may be involved in the pathogenesis of the disease. PMID- 10805469 TI - Additional findings in Galloway-Mowat syndrome. AB - The Galloway-Mowat syndrome consists of early onset nephrotic syndrome and central nervous system anomalies. Additional anomalies including hiatal hernias have been reported but are not consistently present. Various renal histopathologic changes have been reported and it is unclear whether these represent heterogeneity within the syndrome or different stages of disease progression. We describe a patient with Galloway-Mowat syndrome who had a renal biopsy at 9 months that showed minimal changes. She died at 42 months at which time the renal autopsy finding was diffuse mesangial sclerosis. Neuropathologic findings included a small brain with cerebral dysplasia, leukomalacia and absence of the cerebellar granular layer without leptomeningeal heterotopia. The autopsy also revealed bilateral ovarian agenesis. This is the first report of serial histopathologic changes and ovarian agenesis in Galloway-Mowat syndrome. PMID- 10805470 TI - Successful treatment of visceral leishmaniasis in Finnish-type congenital nephrotic syndrome. AB - Visceral leishmaniasis is a parasitic disease endemic in the Mediterranean Basin, including Malta. Finnish-type congenital nephrotic syndrome is an autosomal recessive condition that presents in the first 3 months of life with heavy proteinuria, hypoalbuminemia with secondary edema, and hyperlipidemia. We present an infant with congenital nephrotic syndrome who had had unilateral nephrectomy and who also developed visceral leishmaniasis. He was successfully cleared of the infection by the administration of sodium stibogluconate, with no deterioration of renal function or other complications. PMID- 10805471 TI - Budd-Chiari syndrome and inferior vena cava thrombosis in a nephrotic child. AB - We observed Budd-Chiari syndrome in a boy aged 2 years 6 months with nephrotic syndrome due to hepatic vein and inferior vena cava thrombosis, confirmed by Doppler imaging. Normal values of the routine hemostatic parameters proved that they are of little predictive value for the thrombotic state. Immediate heparin infusion was initiated. High doses of heparin up to 59 IU/kg per hour were required for efficient anticoagulation. A remission of the nephrotic syndrome was achieved with vincristine. Oral anticoagulation with a vitamin K antagonist was continued for 6 months. Doppler imaging then indicated full re-establishment of the blood flow through the affected vessels. The clinical and Doppler data in this case are compatible with acute Budd-Chiari syndrome due to incomplete outflow obstruction of the hepatic veins and inferior vena cava. The favorable outcome was due to the immediate heparin infusion and prompt remission of the nephrotic syndrome. Doppler imaging was an important tool for non-invasive diagnosis and follow-up. PMID- 10805472 TI - Genitourinary complications of systemic lupus erythematosus. AB - A 14-year-old African-American girl was diagnosed with antiphospholipid-positive systemic lupus erythematosus (SLE) in July 1994. The course was complicated by nephrotic syndrome, sepsis, hemolytic anemia, acute renal failure, saphenous vein thrombosis, cutaneous vasculitis, mesenteric vasculitis, appendicitis, hemorrhagic cystitis, and avascular necrosis of the hips. In August 1997, she developed ovarian and fallopian tube complications secondary to SLE. Genitourinary complications of SLE, however, are uncommon, and ovarian vasculitis has not previously been reported as a complication of SLE. This report describes the course of an adolescent patient with SLE and focuses specifically on her genitourinary complications. PMID- 10805473 TI - The emergency management of severe hypertension. AB - Severe hypertension in childhood is a life-threatening clinical problem that carries with it not only serious sequelae of inadequate treatment but equally serious complications of over enthusiastic therapy. The majority of cases have some form of underlying renal disease. Therapeutic success is achieved by slow and safe reduction of blood pressure with the immediate target of avoiding hypertensive sequelae yet allowing preservation of target organ function. Short acting parenteral antihypertensives are recommended along with careful blood pressure monitoring to prevent complications arising through loss of autoregulatory control. PMID- 10805474 TI - The zebrafish pronephros: a genetic system for studies of kidney development. AB - The zebrafish, as a model system for vertebrate development, offers distinct experimental advantages for studies of organogenesis. The simplicity of the zebrafish pronephros, the feasibility of isolating large numbers of mutants, and the growth in infrastructure for genomics makes the zebrafish an attractive system for the analysis of kidney development. Mutants affecting several aspects of nephrogenesis, including differentiation of the intermediate mesoderm, nephron patterning, epithelial polarity, and angiogenesis, have been isolated. Analysis of mutant phenotypes and the cloning of mutant genes has revealed: (1) a role for bone morphogenetic proteins in patterning the ventral mesoderm, (2) an essential role for the pax2.1 gene in pronephric development, (3) multiple loci required for establishing epithelial polarity in the pronephric duct, (4) a central role for podocytes in directing glomerulogenesis, and (5) 15 loci associated with cystic maldevelopment in the pronephros. The striking similarities of pronephric cell types to those found in higher vertebrates, as well as the conservation of kidney-specific gene expression patterns, suggest that insights gained from studies in zebrafish will be broadly applicable to cell differentiation in the kidney. PMID- 10805475 TI - Paucisymptomatic hemolytic uremic syndrome. PMID- 10805476 TI - Angiotensin converting enzyme inhibition in non-diabetic nephropathy with non nephrotic proteinuria. PMID- 10805477 TI - Absence of correlation between genotype and the severity of diffuse mesangial sclerosis in Denys-Drash syndrome. PMID- 10805478 TI - Periodic re-determination of standard blood pressure percentile tables for children and adolescents and the possible effect of a "secular trend"? PMID- 10805479 TI - 7th International Workshop on Developmental Nephrology 9-11 September 1998, Stockholm, Sweden. Early renal development: a key to the understanding of adult diseases. PMID- 10805480 TI - Immunology, lupus and atheroma. PMID- 10805481 TI - Accelerated atheroma in lupus--background. AB - Observational cohort studies in SLE have led to the description of accelerated atherosclerosis as an important cause of mortality and morbidity in this disease. The clinical observation of coronary artery disease occurring in premenopausal females with SLE gave rise to the concept of the bimodal mortality pattern. This pattern was confirmed in autopsy and epidemiological studies. These studies identified hypercholesterolemia and particularly its persistence in the first three years of disease, hypertension, and lupus itself as important risk factors for the development of accelerated atherosclerosis in these patients. It also became evident that corticosteroid therapy plays an important role in the elevation of plasma lipids while antimalarials resulted in a reduction of plasma cholesterol, LDL, and VLDL, especially in steroid-induced hyperlipidemia. Studies of clinical outcomes for atherosclerotic disease (angina, myocardial infarction) have shown a prevalence of 6-12% in a number of SLE cohorts. However, more sensitive investigations including myocardial perfusion imaging and carotid ultrasound have demonstrated a prevalence of atherosclerotic disease in 40% of patients studied. Further studies of SLE disease process, including immunological factors, may more clearly define the pathogenesis of accelerated atherosclerosis in patients with SLE, and may help elucidate mechanisms of atherosclerosis in the general population. PMID- 10805482 TI - Epidemiology of cardiovascular disease in systemic lupus erythematosus. AB - Awareness of the impact of cardiovascular disease on the late morbidity and mortality in patients with Systemic Lupus Erythematosus (SLE) is increasing. Clinical events secondary to accelerated atherosclerosis have been documented in lupus cohorts across the globe. We review the history and epidemiology of cardiovascular disease in patients with SLE. PMID- 10805483 TI - Detection of coronary artery disease and the role of traditional risk factors in the Hopkins Lupus Cohort. AB - Coronary artery disease (CAD) is a major cause of morbidity and mortality in SLE, including the Hopkins Lupus Cohort. Currently, 9% of the cohort have had clinical evidence (angina or myocardial infarction) of CAD. In our initial prospective study we found that duration of prednisone, hypertension, hyperlipidemia and obesity were risk factors for later CAD. We can now extend that list to include age, male sex, elevated homocysteine, renal insufficiency and antiphospholipid antibodies. Many of the risk factors are amenable to intervention, but the timing of intervention, and the effectiveness of intervention, must be determined. PMID- 10805484 TI - Vascular imaging: changing the face of cardiovascular research. AB - Patients with systemic lupus erythematosus (SLE) are at significant risk for premature cardiovascular disease, now a leading cause of death in this population. Most previous studies have used an overt clinical event to identify cardiovascular disease, likely underestimating the actual prevalence in these patients. Although the rates of myocardial infarction in SLE are high, the actual number of coronary events is low, precluding large clinical trials using a coronary event as the sole outcome. The ability to measure atherosclerosis, a known determinant of coronary heart disease, provides investigators with a desirable surrogate for the clinical cardiac event. With the advent of sensitive imaging techniques to identify subclinical atherosclerosis, we are now better equipped to determine the true prevalence and mechanisms of vascular disease in SLE. In this review, we will discuss several vascular imaging techniques and the current trend away from measuring flow-limiting vessel stenosis toward measuring earlier structural and functional aspects of the vascular system. PMID- 10805485 TI - Normal endothelial cell function. AB - Endothelial cell functions, primarily involving regulated mediator secretion or altered surface protein expression, are vital for normal homeostasis. Endothelial cells secrete the potent vasodilator and anti-platelet agent prostacyclin and nitric oxide, and also the potent vasoconstrictor peptide endothelin-1; they control the selective adhesion and emigration of leukocytes from the bloodstream; and they are the source of circulating von Willebrand factor, tissue plasminogen activator and type 1 plasminogen activator inhibitor. The properties of healthy endothelium ensure that an antithrombotic and anticoagulant balance is maintained in the bloodstream, and provide a tonic vasodilator action that controls blood flow and pressure on a minute-to-minute basis. Disturbances of normal endothelial function are strongly implicated in the pathogenesis of atherosclerosis and autoimmune vasculitic diseases including lupus. PMID- 10805486 TI - The endothelium in atherogenesis. AB - The focus of this article is an overview of the endothelial changes that initiate and perpetuate the process of atherogenesis. The endothelium can undergo a series of changes which allow it to participate in the inflammatory response; this is known as endothelial cell activation (ECA). The five core changes of ECA are loss of vascular integrity; expression of leucocyte adhesion molecules; change in phenotype from antithrombotic to prothrombotic; cytokine production, and upregulation of HLA molecules. The diverse effects of ECA share a common intracellular control mechanism through the activation of the transcription factors including Nuclear Factor kappaB. ECA is an initiating step in atherogenesis. Modified low density lipoproteins are probably the major cause of endothelial cell activation in atherogenesis, and become especially so after oxidation, glycation (in diabetes) or incorporation in immune complexes. In antiphospholipid syndrome (APS), antiendothelial cell antibodies have been detected in up to 67% of patients. In vitro studies suggest that aPL causes ECA and thus lead to speculation that aPL by causing ECA may initiate atherogenesis. Further clinical and in vitro studies are required to address these issues. PMID- 10805487 TI - Lipids, cardiovascular disease and atherosclerosis in systemic lupus erythematosus. AB - Systemic lupus erythematosus is commonly associated with early onset cardiovascular disease and is often associated with hyperlipidaemia. This review examines the evidence for an increased prevalence of both CHD and hyperlipidaemia in SLE and mechanisms by which autoimmunity in SLE could accelerate the progression of atheroma. It postulates how lipid lowering therapies used in cardiological disease might help reduce the incidence of CHD in SLE. PMID- 10805488 TI - Antibodies to oxidised LDL. AB - In prospective studies antibodies to oxidised LDL (low density lipoprotein) have been shown to predict myocardial infarction and progression of carotid atherosclerosis in non-autoimmune subjects. The antibodies to oxidised LDL are crossreactive with antiphospholipid antibodies most likely due to their binding to oxidised phospholipids. The frequent occurrence of these antibodies and their association with arterial thrombosis in patients with SLE and antiphospholipid syndrome suggest their involvement in the development of accelerated atherosclerosis in these patients. Some in vitro studies suggest that antibodies to oxidised LDL may have an atherogenic effect by enhancing the lipid accumulation into macrophages in the atherosclerotic vessels. These antibodies can be considered as markers of the pathogenic determinants of atherosclerosis, such as enhanced lipid oxidation, proinflammatory stage and impaired vasodilatation. PMID- 10805489 TI - Lipoprotein(a) oxidation and autoantibodies: a new path in atherothrombosis. AB - Lipoprotein(a) (Lp(a)) is considered a vascular pathogen of outstanding importance. High plasma levels of this lipoprotein are associated with premature arterial disease; however, the mechanisms involved have not been clarified. The atherosclerotic process is increasingly regarded as a chronic inflammatory reaction in the arterial wall where oxidation-mediated endothelial injury involving modified forms of low-density lipoprotein (LDL) seems to be a key event. Autoimmune pathways are involved in the progression of atherosclerosis and humoral response to oxidatively modified LDL can be considered among these pathways. A number of factors can be encountered in the pathogenesis of the accelerated arterial disease seen in patients with antiphospholipid (Hughes) syndrome (APS) and systemic lupus erythematosus (SLE). Among these, high levels of Lp(a) have been described in both and increasing evidence indicates that patients with antiphospholipid antibodies (aPL) are under oxidative stress. Recent studies suggest that the so-called 'oxidation theory of atherosclerosis' may also be applied to Lp(a). This fact makes this lipoprotein potentially suitable as a target of the immune system and antibodies reacting against oxidatively-modified Lp(a) by malondialdehyde have been recently described in APS and SLE. It is therefore likely that an immune response to the oxidized moiety of Lp(a) might be influential in the pathogenicity of this lipoprotein and, subsequently, of atherosclerosis. PMID- 10805490 TI - Accelerated atheroma and anti-beta2-glycoprotein I antibodies. PMID- 10805491 TI - Sex hormone binding globulins and atherosclerotic risk in systemic lupus. AB - Sex hormone binding globulin (SHBG) is a transport protein in human plasma which regulates the bioavailability of sex hormones, mediates membrane receptor signaling and may affect inflammatory processes, suggesting a regulatory role for this protein in the prevention of atherosclerosis. The current report summarizes literature implicating several members of the SHBG family in the regulation of hormonal and inflammatory processes which may be pertinent to the accelerated atherosclerosis seen in systemic lupus. PMID- 10805492 TI - Autoimmunity in atherosclerosis: lessons from experimental models. AB - The modern view of atherosclerosis is of a chronic inflammatory disorder. In accord with this paradigm, the process of uninhibited influx of fat to the vessel wall results from an 'adequate' response to various forms of injury (i.e. turbulence, infections, modified lipoproteins). This idea has been further extended by several groups, to assume that the atherosclerotic lesion can be the target of an autoimmune mediated attack. According to this hypothesis, the site of initiation of the plaque should bear/express the target autoantigen, whereas concomitantly a respective immune response is generated in the periphery. The examples illuminating this notion are beta2GPI as a target autoantigen, HSP60/65 an oxidized-LDL. Herein we present evidence to support the involvement of autoimmune mechanisms in atherogenesis based on the experience from experimental models and human studies. PMID- 10805493 TI - How accelerated atherosclerosis in SLE has changed our management of the disorder. AB - Basic science insights along with the assessment of risk factors so clearly documented by the Canadian and Hopkins cohorts in earlier papers of this special issue have clearly led to a change in the way rheumatologists manage SLE. This review will highlight specific ways it should alter our day to day treatment of lupus patients. PMID- 10805494 TI - Predictors of weight gain in the Pound of Prevention study. AB - OBJECTIVES: This study examined cross-sectional and prospective relationships between macronutrient intake, behaviors intended to limit fat intake, physical activity and body weight. DESIGN: The overall goal was to identify diet and exercise behaviors that predict and/or accompany weight gain or loss over time. Specific questions addressed included: (a) are habitual levels of diet or exercise predictive of weight change; (b) are habitual diet and exercise levels associated cross-sectionally with body weight; and (c) are changes in diet and exercise associated with changes in body weight over time? PARTICIPANTS: Subjects were a sample of community volunteers (n=826 women, n=218 men) taking part in a weight gain prevention project over a 3-year period. MEASURES: Body weight was measured at baseline and annually over the study period. Self-report measures of diet and exercise behavior were also measured annually. RESULTS: Among both men and women, the most consistent results were the positive association between dietary fat intake and weight gain and an inverse association between frequency of physical activity and weight gain. Individuals who weighed more both ate more and exercised less than those who weighed less. Individuals who increased their physical activity level and decreased their food intake over time were protected from weight gain compared to those who did not. Frequency of high-intensity physical activity was particularly important for both men and women. Additionally, women who consistently engaged in higher levels of moderate physical activity gained weight at a slower rate compared to women who were less active. CONCLUSIONS: Overall results indicated that both cross-sectionally and prospectively, the determinants of weight and weight change are multifactorial. Attention to exercise, fat intake and total energy intake all appear important for successful long term control of body weight. PMID- 10805495 TI - Onset of dieting vs binge eating in outpatients with binge eating disorder. AB - OBJECTIVE: To examine the potential significance of the sequence of the onset of dieting and binge eating in binge eating disorder (BED). DESIGN: BED patients were interviewed and completed a battery of psychometrically well-established measures of current eating behaviors, eating disorder psychopathology, and associated psychological functioning. SUBJECTS: Participants were 98 consecutive outpatients with BED evaluated for a clinical trial. MEASURES: Interview data, self-report measures and measured body weight were examined. RESULTS: Participants who reported that dieting preceded binge eating (DIETfirst, 65%) were compared to those who reported that binge eating preceded their first diet (BINGEfirst, 35%). The study groups did not differ in demography, current or highest body mass index, current eating behaviors or psychopathology, or psychological functioning. The two groups did not differ in age of first diet; however, the BINGEfirst group was significantly younger when first overweight, at onset of binge eating, and at onset of BED diagnosis. The BINGEfirst group reported a higher frequency of being teased about their weight. CONCLUSIONS: A substantial subgroup of BED patients report that binge eating preceded their first diet. This finding, which replicates previous reports for BED and appears higher than that generally reported for bulimia nervosa, may have implication for etiologic models of binge eating. PMID- 10805496 TI - Muscle fiber area distribution of musculus uvulae in obstructive sleep apnea and non-apneic snorers. AB - OBJECTIVE: To determine whether differences exist in the morphology of upper airway muscles between apneic and non-apneic snorers DESIGN: Muscle characteristic analysis in patients undergoing upper airway surgery in a tertiary sleep center. SUBJECTS: 10 non-apneic snorers and 10 sleep apnea hypopnea syndrome (SAHS). MEASUREMENTS: Frequency distribution of musculus uvulae (MU) muscle fiber area determined from 475+/-207 (mean s.d.) and 697 +/- 165 type IIA fibers and 92 +/- 32 and 68 45 type I fibers in snorers and SAHS, respectively. RESULTS: Histochemical analyses of MU revealed as expected that type IIA fibers occupied a significantly larger area within that muscle in SAHS compared to Snorers (89.4 +/- 5.8% and 76.1 +/- 15.1% respectively, P= 0.01). No difference was found in the frequency distribution of type I and type IIA fiber areas between the two groups comparing the between-groups and within-group variance of individual area distributions found in snorers and SAHS. CONCLUSION: There is no difference in muscle fiber area frequency distribution between non-apneic snorers and SAHS patients. This suggests that musculus uvulae of these groups of patients does not have a specific prevalence of atrophic or hypertrophic muscle fibers. PMID- 10805497 TI - Food-induced cortisol secretion in relation to anthropometric, metabolic and haemodynamic variables in men. AB - OBJECTIVE: To assess the relationships between the regulation of diurnal and food induced cortisol secretion and anthropometric, metabolic and haemodynamic variables in middle-aged men. SUBJECTS AND METHOD: Salivary cortisols were collected repeatedly (n = 7) over an ordinary working day (8a.m. to 11 p.m.) in a randomly selected population of 284 men, aged 51 y. A standardized lunch was provided, and an overnight low-dose dexamethasone suppression test was performed. These measurements were correlate with the anthropometric factors-body mass index, BMI, (kg/m2), waist-to-hip ratio (WHR) and abdominal sagittal diameter; the metabolic factors-fasting insulin and glucose as well as their ratio, and triglycerides; and the haemodynamic factors-systolic and diastolic blood pressures and heart rate. RESULTS: As reported previously two principal types of salivary cortisol secretory patterns can be singled out, one characterized by high morning cortisol levels, a normal circadian rhythm and feedback regulation (dexamethasone) along with a brisk cortisol response to lunch, and another, found in a limited number of men, characterized by low morning cortisols, the absence of a circadian rhythm, a relative resistance to dexamethasone inhibition and a poor lunch-induced cortisol response. The normal cortisol secretory pattern showed negative associations with BMI (P< 0.05), WHR (P< 0.01), and blood pressures (P< 0.001). After stimulation by food intake, negative relationships were found with all obesity measurements, insulin, insulin/glucose ratio, triglycerides, blood pressures and heart rate (all P < 0.001). These results suggest that normally regulated cortisol is associated with a favourable somatic health. In contrast, after food intake cortisol secretion, based on an abnormal cortisol secretory pattern, showed consistent positive associations with obesity measurements, insulin, glucose and insulin/glucose ratio, triglycerides, blood pressures and heart rate (all P < 0.001). CONCLUSIONS: A normal HPA axis regulation is associated with excellent health anthropometric, metabolic and haemodynamic variables, particularly visible after the physiological stimulus of food intake. This is, however, not the case in men with perturbed HPA axis function where associations in these somatic variables become exaggerated by food intake. We have previously reported that perceived stress-related cortisol is associated with abnormalities in the variables mentioned above, both with a normal and, particularly, with an abnormal function of the HPA axis. It is thus apparent that perceived stress and food intake show separate associations to somatic variables with a normally functioning HPA axis, while with an abnormal regulation of this axis both perceived stress and food intake exaggerate associations to abnormal somatic variables. PMID- 10805498 TI - In vivo effects of CGP-12177 on the expression of leptin and uncoupling protein genes in mouse brown and white adipose tissues. AB - OBJECTIVE: To assess the effect of chronic treatment with CGP-12177 a beta3 adrenergic receptor (AR) agonist with beta2/beta1-AR antagonist action, on the expression of the leptin gene and of genes coding for uncoupling proteins (ucp1, ucp2 and ucp3) in brown and white adipose tissues. DESIGN: NMRI mice received a daily subcutaneous injection of CGP-12177 at a dose of 0.05, 0.2, 0.5 or 1 mg/kg for 15 days. The specific levels of the mRNAs of interest were analysed in interscapular brown adipose tissue (BAT) and in two white adipose tissue (WAT) depots, inguinal (IWAT) and epididymal (EWAT). RESULTS: No changes in food intake or body weight were detected at any dose of CGP-12177. In the two WAT depots, the treatment led to enhanced expression of ucp1 and ucp3, but not of ucp2. In BAT, low doses (0.05 and 0.2 mg/kg) led to a decreased expression of the three ucp genes, whereas a slight stimulatory effect on the three ucp genes was elicited with a high dose (1 mg/kg). Treated animals displayed increased expression of leptin in BAT and, to a lesser extent, in IWAT, but not in EWAT. CONCLUSION: The results reveal that simultaneous stimulation of the expression of certain ucp genes and the leptin gene can be achieved, and suggest that adrenergic regulation of the leptin gene and of genes of the ucp family in adipose tissues is the result of complex interactions between the different beta-AR pathways. PMID- 10805499 TI - Activity related energy expenditure in children and adolescents with Prader-Willi syndrome. AB - OBJECTIVE: To measure activity related energy expenditure in Prader-Willi syndrome (PWS) corrected for body size. SUBJECTS: 17 PWS subjects (10 females, seven males, age 7.5-19.8 y) and 17 obese controls, matched for gender and bone age. MEAUREMENTS: Basal metabolic rate (BMR) was measured by ventilated hood and average daily metabolic rate (ADMR) was measured with doubly labelled water. Activity induced energy expenditure (AEE) was calculated as 0.9ADMR -BMR. Activity related energy expenditure was corrected for body size using the following measures: AEE per kg body weight (AEE/kg), ADMR/BMR (PAL), and the residual of the regression of ADMR on BMR (rADMR). Group differences were analyzed by analysis of covariance adjusting for bone age, fat mass (FM) and gender. RESULTS: ADMR, AEE and PAL were lower (P < 0.01) in the PWS group compared with the control group (7.14 +/- 1.72, 1.07 +/- 0.69 and 1.33 +/- 0.15 MJ/day compared with 9.94 +/- 2.64, 2.56 +/- 1.03 and 1.55 +/- 0.12 MJ/day respectively). The variance of AEE/kg and PAL was significantly explained by gender and PWS, while AEE was additionally explained by FM. The variance of rADMR was explained by PWS and not by FM or gender. CONCLUSION: Activity related energy expenditure is decreased in PWS compared with controls adjusted for bone age, FM and gender. PMID- 10805500 TI - Do we eat less fat, or just report so? AB - OBJECTIVE: To examine secular trends in diet reporting error. METHODS: Dietary information was obtained from 228 Danish men and women in 1987-88, and from 122 men and women in 1993-94. RESULTS: Bias in dietary reporting of energy and protein intake was assessed by comparing reported intake with intake data, estimated from 24 h nitrogen output, validated by administering P-aminobenzoic acid, and estimated 24 h energy expenditure. Total energy was under-reported more than energy from protein at both surveys, suggesting that energy from other nutrients, like fat and/or carbohydrate, must have been under-reported too. There was a greater under-reporting for energy than for protein in 1993-94 (29%) than in 1987-88 (15%). Obesity was positively associated with under-reporting, both in 1987-88 and in 1993-94. CONCLUSION: The higher macro-nutrient specific error in 1993-94 compared to 1987-88 may reflect a trend to increasingly omitting fat and/or carbohydrate-rich foods in dietary reporting. This may be a consequence of increased awareness of diet intake, which, in turn, may be related to intensified public health campaigns to reduce intake of fat and/or simple carbohydrate. These results may have consequences for our understanding of the apparent decline in dietary fat and associated health benefits. PMID- 10805502 TI - Leptin is influenced both by predisposition to obesity and diet composition. AB - OBJECTIVE: (1) To investigate whether plasma leptin concentrations differ between subjects with and without the genetic predisposistion to obesity, and (2) to investigate the effect of dietary manipulations on plasma leptin in these subjects. DESIGN: Fasting and postprandial plasma leptin concentrations were measured before and after 14 days' ad libitum intake of a fat-rich (FAT), starch rich (STARCH) or sucrose-rich (SUCROSE) diet. On day 15 ad libitum breakfast and lunch were given and blood sampled regularly until 6 p.m. SUBJECTS: Eight normal weight, post-obese women and 10 matched controls (body mass index, 23.5+/-0.5 and 22.9 +/- 0.3 kg/m2). MEASUREMENTS: Leptin, glucose, insulin, appetite ratings, dietary intake, body weight and composition. RESULTS: Fasting leptin concentration on day 1 or 15 did not differ between post-obese and controls. However, after meal intake leptin increased in post-obese compared with controls on all three diets. In both groups fasting and postprandial leptin concentrations were greater after SUCROSE compared with FAT and STARCH. CONCLUSION: A larger postprandial leptin concentration was observed in post-obese subjects than in controls. This may be related to greater insulin sensitivity in adipose tissue in the post-obese. Furthermore, increased leptin concentrations were found after a sucrose-rich diet in both groups, possibly related to larger postprandial insulin peaks on this diet. Both contentions should, however, be validated by further studies. PMID- 10805501 TI - Association of Trp64Arg polymorphism of the beta3-adrenergic receptor gene and no association of Gln223Arg polymorphism of the leptin receptor gene in Japanese schoolchildren with obesity. AB - OBJECTIVE: To investigate whether Trp64Arg polymorphism of the beta3-adrenergic receptor (beta3-AR) gene and Gln223Arg polymorphism of the leptin receptor (Ob-R) gene are associated with obesity in Japanese schoolchildren. DESIGN: Population study of participants from a rural town located within 50 km northeast of Tokyo based on school medical examinations. SUBJECTS: 553 Japanese schoolchildren (291 boys and 262 girls) who were 9-15 y old with a mean age of 11.9 +/- 1.8 y. MEASUREMENTS: DNA was extracted from whole blood and genotyped by PCR-RFLP. Height, weight and blood pressure were measured in school medical examinations. Total cholesterol, triglyceride and HDL-cholesterol concentrations were measured by an autoanalyzer. Obesity index, body mass index (BMI) and LDL-cholesterol concentration were calculated by the respective formulae. RESULTS: In Trp64Arg polymorphism of the beta3-AR gene, the number of obese subjects with Trp/Arg or Arg/Arg genotypes was significantly higher than that of the non-obese subjects (chi2=5.79, P=0.02). The obesity index of subjects with the Arg/Arg or Arg/Trp genotype was significantly higher than that of those with the Trp/Trp genotype (8.2 +/- 18.7% vs 4.5 +/- 15.8%, P=0.04). Moreover, after adjustments for age and gender, BMI of subjects with the Trp/Arg or Arg/Arg genotype was significantly higher than that of those with the Trp/Trp genotype (19.4 +/- 3.6 kg/m2 vs 18.9 +/- 3.2 kg/m2, P= 0.02). However, no significant differences were observed in the clinical characteristics among the genotype groups of the Ob-R gene. CONCLUSIONS: Trp64Arg polymorphism of the beta3-AR gene appears to be a genetic risk factor for obesity in Japanese children, but Gln223Arg polymorphism of the Ob-R gene does not appear to be associated with obesity. PMID- 10805503 TI - The genetic background modifies the effects of the obesity mutation, 'fatty', on apolipoprotein gene regulation in rat liver. AB - BACKGROUND: Obesity is associated with disorders of plasma lipid transport in many, but not in all obese subjects. The effects of obesity on the regulation of genes involved in plasma lipid transport may depend on specific mutations causing or contributing to obesity and/or on interactions of a specific obesity mutation with the genetic background. The 'fatty' (Glu269Pro) leptin receptor mutation causes severe obesity associated with hypertriglyceridaemia and altered hepatic apolipoprotein gene regulation in Zucker fatty rats. OBJECTIVE: To determine whether the effects of the obesity mutation 'fatty' on apolipoprotein gene regulation in rat liver depend on the genetic background. METHODS: We studied hepatic apolipoprotein (apo) A-IV, A-I, and C-III gene expression in obese rats carrying the 'fatty' mutation on the background of the Zucker or Wistar strain. RESULTS: Basal apoA-IV gene expression was increased in fatty rats of both strains, whereas apoA-I and apoC-III gene expression differed between Wistar and Zucker fatty rats: apoA-I gene transcription was reduced to half and apoC-III mRNA was increased two-fold in Wistar fatty, but not in Zucker fatty rats vs lean controls. A fish oil diet suppressed apoA-IV, but not apoA-I gene transcription in Wistar fatty rats, whereas in Zucker fatty rats apoA-IV transcription was unaffected, but apoA-I transcription was suppressed. CONCLUSIONS: Interactions of the 'fatty' leptin receptor mutation with the genetic background significantly affect the basal and diet-induced regulation of the apoA-IV, C-III and A-I genes in rat liver. The genetic background may therefore be a major determinant of the consequences of a specific obesity mutation for plasma lipid transport. PMID- 10805504 TI - Relationship between altered postprandial lipemia and insulin resistance in normolipidemic and normoglucose tolerant obese patients. AB - OBJECTIVE: Although there are changes in the postprandial lipid responses of obese patients, these are closely associated with high fasting triglycerides (TG). This study of 17 normotriglyceridemic, normoglucose-tolerant android obese subjects (body mass index, BMI = 34.3 +/- 3.1 kg/m2) and 33 normal-weight controls (BMI = 21.8 +/- 1.6 kg/m2) was done to examine their postprandial responses to an oral fat loading test containing retinol (890 calories, 85% fat) and to evaluate the possible association between clinical and biological features of obesity and/or insulin resistance and postprandial lipemia. SUBJECTS AND MEASUREMENTS: Blood samples were taken before giving the fat load and at 2,3,4,5,6 and 8 h after it. Insulin sensitivity was assessed using HOMA, and TG and retinyl palmitate (RP) in the plasma, chylomicrons and non-chylomicron fractions were measured each time. RESULTS: The areas under the curves (AUC) of chylomicron TG for the obese and controls were not different, indicating adequate lipolytic activity. By contrast, the AUC for non-chylomicron TG was significantly greater in the obese than in the controls (512 +/- 135 vs 429 +/- 141 mmol/lmin, P < 0.01). In addition, the AUC for RP in this same fraction was significantly lower in the obese than in the controls (103 +/- 55 vs 157 +/- 88 mg/l min, P < 0.05), suggesting that the TG from endogenous lipoproteins accounted for most of the increase in TG in the non chylomicron fraction. Parameters related to obesity showed no relationship with these postprandial abnormalities, whereas HOMA, which discriminated between the groups, partly explained (r2= 23%, P < 0.01) the significant increase in non-chylomicron TG. CONCLUSIONS: Android obese patients with a fasting TG in the normal range and not different from the fasting TG of lean controls had an abnormal postprandial lipemia response, indicated by a significantly greater TG in the non-chylomicron subfraction than in controls. These alterations may be partly due to postprandial changes in endogenous lipoproteins as a consequence of insulin resistance. PMID- 10805505 TI - Impact of birth weight on ethnic variations in subcutaneous and central adiposity in American children aged 5-11 years. A study from the Third National Health and Nutrition Examination Survey. AB - OBJECTIVE: To determine the types of subcutaneous adiposity represented by different measurements of skinfold thickness that are associated with birth weight in white (n=759), Black (n=916) and Hispanic (n=813) American children aged 5-11 y. We also determined the contribution of birth weight to ethnic differences in subcutaneous and central adiposity. DESIGN AND METHODS: Data for this analysis were from the Third US National Health and Nutrition Examination Survey. The outcome measures were triceps, subscapular, suprailiac and thigh skinfold thicknesses at 5-11 y of age. Central adiposity was defined as ratios of subscapular to triceps (STR) and central-peripheral (CPR) (subscapular+suprailiac)/(triceps+thigh) skinfolds. Partial correlation analyses were used to determine the association between birth weight and measures of subcutaneous fatness, while multiple linear regression analyses were used to determine the independent contribution of birth weight to ethnicity variations in subcutaneous and central adiposity adjusting for sex, age and BMI. RESULTS: Overall, birth weight was negatively associated with subscapular skinfold and central adiposity in White, Black and Hispanic American children (P < 0.05). Birth weight was also negatively associated with suprailliac skinfold in both Blacks and Hispanics (P < 0.01) and with sum of the four skinfolds in Blacks (P < 0.05). Compared with White, Black ethnicity was negatively associated with triceps, suprailliac thigh and sum of skinfold thicknesses controlling for birth weight, sex, age and BMI (P < 0.01). Compared with White, Hispanic ethnicity was negatively associated with triceps, thigh and sum of skinfold thicknesses (P < 0.01). Both Black and Hispanic ethnicity was positively associated with STR and CPR (P < 0.01). CONCLUSIONS: In this population of American children, the association of birth weights with subcutaneous and central fat accumulation may be due to fetal programming. Since the impact of fetal conditions is likely to be modified by life course, defining the interaction between factors that are present at birth and subsequent exposures is one of the essential challenges for future research. PMID- 10805506 TI - Low serum testosterone level as a predictor of increased visceral fat in Japanese American men. AB - OBJECTIVE: To examine the association between baseline testosterone levels and changes in visceral adiposity in Japanese-American men. DESIGN: Prospective observational study. SUBJECTS: Second-generation Japanese-American males enrolled in a community-based population study. MEASUREMENTS: At baseline, 110 men received a 75g oral glucose tolerance test (OGTT), and an assessment of body mass index (BMI); visceral adiposity measured as intra-abdominal fat area (IAF) using computed tomography (CT); fasting insulin and C-peptide levels; and total testosterone levels. IAF was re-measured after 7.5 y. Subcutaneous fat areas were also measured by CT in the abdomen, thorax and thigh. The total fat (TF) was calculated as the sum of IAF and total subcutaneous fat areas (SCF). RESULTS: After 7.5y, IAF increased by a mean of 8.0 cm2 (95% CI: 0.8, 15.3). Baseline total testosterone was significantly correlated with change in IAF (r= -0.26, P= 0.006), but not to any appreciable degree with change in BMI, TF, or SCF. In a linear regression model with change in IAF as the dependent variable, baseline testosterone was significantly related to this outcome while adjusting for baseline IAF, SCF, BMI, age, diabetes mellitus status (OGTT by the WHO diagnostic criteria) and fasting C-peptide (regression coefficient for baseline testosterone [nmol/l] = -107.13, P = 0.003). CONCLUSIONS: In this Japanese-American male cohort, lower baseline total testosterone independently predicts an increase in IAF. This would suggest that by predisposing to an increase in visceral adiposity, low levels of testosterone may increase the risk of type 2 diabetes mellitus. PMID- 10805507 TI - Similar weight loss with low-energy food combining or balanced diets. AB - OBJECTIVE: The goal of this study was to evaluate the effect of two diets ('food combining' or dissociated vs balanced) on body weight and metabolic parameters during a 6-week period in an in-hospital setting. SUBJECTS AND DESIGN: 54 obese patients were randomly assigned to receive diets containing 4.5 MJ/day (1100 kcal/day) composed of either 25% protein, 47% carbohydrates and 25% lipids (dissociated diet) or 25% protein, 42% carbohydrates and 31% lipids (balanced diet). Consequently, the two diets were equally low in energy and substrate content (protein, fat and carbohydrate) but widely differed in substrate distribution throughout the day. RESULTS: There was no significant difference in the amount of weight loss in response to dissociated (6.2 +/- 0.6 kg) or balanced (7.5 +/- 0.4 kg) diets. Furthermore, significant decreases in total body fat and waist-to-hip circumference ratio were seen in both groups, and the magnitude of the changes did not vary as a function of the diet composition. Fasting plasma glucose, insulin, total cholesterol and triacylglycerol concentrations decreased significantly and similarly in patients receiving both diets. Both systolic and diastolic blood pressure values decreased significantly in patients eating balanced diets. The results of this study show that both diets achieved similar weight loss. Total fat weight loss was higher in balanced diets, although differences did not reach statistical significance. Total lean body mass was identically spared in both groups. CONCLUSION: In summary at identical energy intake and similar substrate composition, the dissociated (or 'food combining') diet did not bring any additional loss in weight and body fat. PMID- 10805508 TI - Sagittal abdominal diameter compared with other anthropometric measurements in relation to cardiovascular risk. AB - BACKGROUND: Abdominal adiposity has been described as an independent risk factor for coronary heart disease. Sagittal abdominal diameter has been found to be closely related to the amount of visceral adipose tissue. AIM: To compare the sagittal abdominal diameter with other anthropometric measures regarding their relationships to risk factors for coronary heart disease (CHD). DESIGN: A study of 885 men and women participating in a health survey. MEASUREMENTS: Sagittal abdominal diameter, body mass index (BMI), waist and hip circumferences, waist-to hip ratio, serum concentrations of risk factors for CHD, blood pressure. RESULTS: In men the sagittal abdominal diameter showed stronger correlations to the CHD risk factors serum cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, glucose, insulin, apolipoprotein B (apoB), plasminogen activator inhibitor tissue-type plasminogen activator (t-PA) and lipid-corrected alpha tocopherol, and to systolic and diastolic blood pressures than the other anthropometric measurements. In women, compared with the other anthropometric measurements the sagittal abdominal diameter was more strongly correlated to serum cholesterol, LDL cholesterol, LDL/HDL (high-density lipoprotein), apo B and t-PA, and to systolic and diastolic blood pressure. The sagittal abdominal diameter showed a stronger correlation to 'total risk' for cardiovascular disease (+ 0.66 for men, 0.62 for women), than waist circumference (+ 0.63 for men, + 0.57 for women) and waist-to-hip ratio (+ 0.61 for men and +0.48 for women; P <0.0001 for all correlations). This diameter was also more strongly correlated to 'metabolic risk' (+ 0.64 for men, + 0.59 for women) than waist circumference (+ 0.60 for men, + 0.59 for women) and waist-to-hip ratio (+ 0.58 for men, + 0.52 for women)(P < 0.0001 for all correlations). In a regression analysis including the anthropometric measurements and the risk values, the sagittal diameter was the strongest measure of cardiovascular risk in both men and women. CONCLUSIONS: Among both men and women in this study the sagittal abdominal diameter showed stronger correlations to cardiovascular risk and to other risk factors in the metabolic syndrome than other anthropometric variables such as waist circumference, waist-to hip ratio and BMI. PMID- 10805509 TI - Preperitoneal fat thickness determined by ultrasonography is correlated with coronary stenosis and lipid disorders in non-obese male subjects. AB - OBJECTIVE: To investigate the relationship between preperitoneal fat thickness (PFT) determined by ultrasonography and the risk of coronary arterial disease, 130 non-obese patients with ischemic heart disease (77 men and 53 women) were examined. RESULTS: There was a positive correlation between PFT and coronary artery stenosis score (r = 0.212, P < 0.05). After dividing the patients by gender, the correlation was recognized only in men (r = 0.246, P< 0.05). Also, PFT was positively correlated to serum total cholesterol (r = 0.259, P < 0.01), triglyceride (r = 0.205, P < 0.05) and low density lipoprotein (LDL)-cholesterol (r = 0.205, P < 0.05), and negatively correlated to serum high density lipoprotein (HDL)-cholesterol (r = -0.261, P < 0.01). Again, these correlations were found only in men, not in women. CONCLUSION: PFT shows good correlations with coronary artery stenosis score and dyslipidemia, and may lead to the development of coronary artery disease in non-obese male subjects. PMID- 10805510 TI - The postprandial rates of glycogen and lipid synthesis of lean and obese female Zucker rats. AB - OBJECTIVE: To determine the relative rates of glycogenesis and lipogenesis following administration of a test meal in lean and obese Zucker rats. PROTOCOL: Nine-week-old lean and obese Zucker rats were fasted overnight, then tube-fed a test meal of balanced composition amounting to 16kJ (lean rats and one group of obese rats) or 24kJ (one group of obese rats) and containing 200 mg 1-(13)C glucose. Immediately after the meal the rats were injected intraperitoneally with 5 mCi of 3H2O and killed 1 h later. METHODS: Glycogenesis was calculated from the incorporation of 3H into liver glycogen divided by the specific activity of plasma water. Lipogenesis was calculated similarly from the incorporation of 3H into saponifiable lipids in liver and perirenal adipose tissue. The proportion of glycogen synthesized by the indirect pathway via pyruvate was determined from the ratio of 3H labelling at positions C6 and C2 in the glycogen glucose residues. Glycogen synthesis from glucose was determined from the ratio of 13C enrichment in liver glycogen to that in plasma glucose. RESULTS: The rate of synthesis of glycogen was considerably lower in the livers of obese rats than those of lean controls, with the larger meal causing a small but significant increase in glycogenesis. The proportion of glycogen synthesized via pyruvate showed a non significant increase in the obese rats, while the amount of glycogen synthesized from glucose was significantly decreased. Hepatic lipogenic rates were about five times higher in both groups of obese rats than the lean controls. In adipose tissue, lipogenesis per g tissue was slightly reduced in the obese rats, although there was clearly an increase in adipose tissue lipogenic activity per whole animal. The larger meal caused a greater rise in plasma glucose and insulin concentrations but did not affect lipogenic rates, although it did cause a greater suppression of lipolysis, as indicated by a lower plasma glycerol concentration. CONCLUSION: Ingested carbohydrate is partitioned predominantly into hepatic fatty acid synthesis in obese Zucker rats. Hepatic glycogen synthesis is suppressed and comes mainly from precursors other than glucose. The suppression of hepatic glycogen synthesis may contribute to the increased energetic efficiency of obese Zucker rats. PMID- 10805511 TI - Synergistic effect of polymorphisms in uncoupling protein 1 and beta3-adrenergic receptor genes on long-term body weight change in Finnish type 2 diabetic and non diabetic control subjects. AB - OBJECTIVE: To investigate the independent and combined effects of the Trp64Arg polymorphism of the beta3-adrenergic receptor (beta3AR) gene and the (-3826) A- >G polymorphism of the uncoupling protein 1 (UCP1) gene on body weight change in type 2 diabetic and non-diabetic control subjects during a 10y follow-up study. DESIGN: Controlled 10y follow-up study with baseline, 5 and 10y examinations. SUBJECTS: 70 newly diagnosed, middle-aged type 2 diabetic patients and 123 non diabetic control subjects from eastern Finland. MEASUREMENTS: Anthropometric measurements, blood pressure, oral glucose tolerance test, plasma insulin, plasma C-peptide and HbA1c. Genotypes by polymerase chain reaction followed by enzymatic digestion. RESULTS: No significant differences were found in the frequencies of the two polymorphisms between diabetic and control subjects. The polymorphisms were not cross-sectionally or longitudinally associated with body weight or BMI in diabetic or control subjects. When the diabetic and control subjects were analysed together, the change in the mean body weight was significantly greater among the subjects with both polymorphisms (n = 11) than among those with no polymorphisms (n = 103; change in weight 6.5 +/- 2.5% vs -0.2 +/- 0.8%, P=0.036, and change in Body Mass Index 8.5 +/- 2.6% vs 2.0 +/- 0.8%, P= 0.060, mean +/- s.e.m.). CONCLUSIONS: The simultaneous existence of the two polymorphisms was associated with a tendency to gain weight suggesting a synergistic effect of these polymorphisms on body weight gain. PMID- 10805512 TI - A polymorphism in the gene encoding CART is not associated with obesity in Pima Indians. AB - Cocaine- and amphetamine-regulated transcript (CART) is a hypothalamic peptide that may be involved in the regulation of food intake. We screened the human CART gene and identified a novel C-->G substitution in the 3'-untranslated region of exon 3. This polymorphism was genotyped in a total of 68 Pima Indians with extremes of BMI. The frequencies for alleles C and G were 0.76 and 0.24, respectively. There was no evidence of an association between the genotype frequency at this variant and extremes of BMI in this group of Pima Indians. The results of this study do not suggest a significant role for this variant in exon 3 of CART as a determinant of obesity in Pima Indians. PMID- 10805513 TI - The Pro12Ala variant of peroxisome proliferator-activated receptor-gamma2 (PPAR gamma2) is associated with measures of obesity in Mexican Americans. AB - Peroxisome proliferator-activated receptors (PPAR) are transcription factors that regulate adipocyte differentiation and gene expression. We tested the hypothesis that the Pro12Ala variant of PPAR-gamma2 is associated with obesity and type 2 diabetes-related traits in 921 subjects from the San Antonio Family Heart Study. Subjects with at least one Ala allele (n=210) had significantly higher body mass index (P=0.015) and waist circumference (P=0.028) and significantly higher levels of serum leptin (P= 0.022) than those without the allele. Further studies will determine whether the Pro12Ala variant itself, or other genetic variation at PPAR gamma, is responsible for this association with measures of obesity in Mexican Americans. PMID- 10805514 TI - Two-dimensional electrophoresis of membrane proteins using immobilized pH gradients. AB - Two-dimensional electrophoresis (2-DE) is a highly resolving technique for arraying proteins by isoelectric point and molecular mass. To date, the resolving ability of 2-DE for protein separation is unsurpassed, thus ensuring its use as the fundamental separation method for proteomics. When immobilized pH gradients (IPGs) are used for isoelectric focusing in the first dimension, excellent reproducibility and high protein load capacity can be achieved. While this has been beneficial for separations of soluble and mildly hydrophobic proteins, separations of membrane proteins and other hydrophobic proteins with IPGs have often been poor. Stimulated by the growing interest in proteomics, recent developments in 2-DE methodology have been aimed at rectifying this situation. Improvements have been made in the area of protein solubilization and sample fractionation, leading to a revamp of traditional approaches for 2-DE of membrane proteins. This review explores these developments. PMID- 10805515 TI - Screening assay for promigratory/antimigratory compounds. AB - Large-scale screening strategies aimed at finding anticancer drugs traditionally focus on identifying cytotoxic compounds that attack actively dividing cells. Because progression to malignancy involves acquisition of an aggressively invasive phenotype in addition to hyperproliferation, simple and effective screening strategies for finding compounds that target the invasive aspects of cancer progression may prove valuable for identifying alternative and preventative cancer therapies. Here, we describe a fluorescence-based automated assay for identifying antimigratory compounds, with the ability to discern cytotoxic from noncytotoxic modes of action. With this assay, we analyzed the effects of two drugs on tumorigenic (MDA-MB-435) and nontumorigenic (MCF-10A) human breast cell lines. We chose to compare carboxyamidotriazole (CAI), an experimental compound shown to inhibit migration of various cell types, with tamoxifen, a common preventative and therapeutic anticancer compound. Our assay demonstrated that both these compounds inhibit migration at sublethal concentrations. Furthermore, CAI was more effective than tamoxifen at inhibiting chemotactic and haptotactic migration of both cell lines at all concentrations tested. PMID- 10805517 TI - Characterization of the surfaces generated by liposome binding to the modified dextran matrix of a surface plasmon resonance sensor chip. AB - The dextran matrix of a surface plasmon resonance (SPR) sensor chip modified with hydrophobic residues (BIAcore sensor chip L1) provides an ideal substrate for liposome adsorption. Liposomes of different lipid compositions are captured on the sensor chips by inserting these residues into the liposome membrane, thereby generating stable lipid surfaces. To gain a more detailed understanding of these surfaces, and to prove whether the liposomes stay on the matrix as single particles or form a continuous lipid layer by liposome fusion, we have investigated these materials, using atomic force microscopy (AFM) and fluorescence microscopy. Force measurements with AFM probes functionalized with bovine serum albumin (BSA) were employed to recognize liposome adsorption. Analysis of the maximal adhesive force and adhesion energy reveals a stronger interaction between BSA and the dextran matrix compared to the lipid-covered surfaces. Images generated using BSA-coated AFM tips indicated a complete and homogeneous coverage of the surface by phospholipid. Single liposomes could not be detected even at lower lipid concentrations, indicating that the liposomes fuse and form a lipid bilayer on the dextran matrix. Experiments with fluorescently labeled liposomes concurred with the AFM studies. Surfaces incubated with liposomes loaded with TRITC-labeled dextran showed no fluorescence, indicating a complete release of the encapsulated dye. In contrast, surfaces incubated with liposomes containing a fluorescently labeled lipid showed fluorescence. PMID- 10805516 TI - Generation of mammalian cells stably expressing multiple genes at predetermined levels. AB - Expression of cloned genes at desired levels in cultured mammalian cells is essential for studying protein function. Controlled levels of expression have been difficult to achieve, especially for cell lines with low transfection efficiency or when expression of multiple genes is required. An internal ribosomal entry site (IRES) has been incorporated into many types of expression vectors to allow simultaneous expression of two genes. However, there has been no systematic quantitative analysis of expression levels in individual cells of genes linked by an IRES, and thus the broad use of these vectors in functional analysis has been limited. We constructed a set of retroviral expression vectors containing an IRES followed by a quantitative selectable marker such as green fluorescent protein (GFP) or truncated cell surface proteins CD2 or CD4. The gene of interest is placed in a multiple cloning site 5' of the IRES sequence under the control of the retroviral long terminal repeat (LTR) promoter. These vectors exploit the approximately 100-fold differences in levels of expression of a retrovirus vector depending on its site of insertion in the host chromosome. We show that the level of expression of the gene downstream of the IRES and the expression level and functional activity of the gene cloned upstream of the IRES are highly correlated in stably infected target cells. This feature makes our vectors extremely useful for the rapid generation of stably transfected cell populations or clonal cell lines expressing specific amounts of a desired protein simply by fluorescent activated cell sorting (FACS) based on the level of expression of the gene downstream of the IRES. We show how these vectors can be used to generate cells expressing high levels of the erythropoietin receptor (EpoR) or a dominant negative Smad3 protein and to generate cells expressing two different cloned proteins, Ski and Smad4. Correlation of a biologic effect with the level of expression of the protein downstream of the IRES provides strong evidence for the function of the protein placed upstream of the IRES. PMID- 10805518 TI - 2-Methoxy-4-(2-phthalimidinyl)phenylsulfonyl chloride as a fluorescent labeling reagent for determination of phenols in high-performance liquid chromatography and application for determination of urinary phenol and p-cresol. AB - Highly sensitive fluorescent labeling reagents, 2-(alkyloxy)-4-(2 phthalimidinyl)phenylsulfonyl chlorides (alkyloxy = methoxy, ethoxy, and propoxy; MPS-Cl, EPS-Cl, and PPS-Cl, respectively), for determination of phenols by high performance liquid chromatography (HPLC) have been developed. These reagents react with phenols in basic medium to produce the corresponding fluorescent sulfonyl esters. The maximum fluorescence wavelengths of the derivatives were 308 nm for excitation and 410 nm for emission. The peaks due to phenol labeled with MPS-Cl, EPS-Cl, and PPS-Cl eluted at 6.3, 8.8, and 12.8 min, respectively, on a reversed-phase column with isocratic elution of methanol/water (2:1, v/v), and the detection limits (signal-to-noise = 3) of the derivatives were 10, 12, and 17 fmol per injection, respectively. Among these reagents, MPS-Cl was advantageous because its derivatives had shorter retention times and higher sensitivities in HPLC. The efficiency of converting phenol to the fluorescent derivative by MPS-Cl was about 100%. When MPS-Cl was applied to the determination of urinary phenol and p-cresol by HPLC using p-ethylphenol as an internal standard, the derivatives were separated at retention times of 6.3, 8.7, and 12.3 min, respectively, under the HPLC conditions described above. The concentrations of phenol and p-cresol in normal human urine were 11.9-293.5 and 8.2-346.1 nmol/mg creatinine, respectively. PMID- 10805519 TI - A fluorometric assay for L-asparaginase activity and monitoring of L-asparaginase therapy. AB - The antineoplastic enzyme L-asparaginase is commonly used for the induction of remission in acute lymphoblastic leukemia (ALL). There is no simple method available for measuring the activity of this highly toxic drug. We incubated L asparaginase from Erwinia chrysanthemi with L-aspartic acid beta-(7-amido-4 methylcoumarin) and measured the release of 7-amino-4-methylcoumarin fluorometrically for 30-300 min. The rate of the hydrolysis of the substrate was linear over a 50-fold range of the concentration of the enzyme. With increasing substrate concentration, the enzyme showed a saturable kinetic pattern with V(max) of 0.547 (SD 0.059) microM/min/mg of enzyme (n = 3) and Km of 0.302 (SD 0.095) mM (n = 3). This assay enables rapid analysis of L-asparaginase activity in biological samples and it can be used, for example, for monitoring of L asparaginase activity in serum of ALL patients during their L-asparaginase therapy. PMID- 10805520 TI - Enhanced prediction accuracy of protein secondary structure using hydrogen exchange Fourier transform infrared spectroscopy. AB - A novel equilibrium hydrogen exchange Fourier transform IR (HX-FTIR) spectroscopy method for predicting secondary structure content was employed using spectra obtained for a training set of 23 globular proteins. The IR bandshape and frequency changes resulting from controlled levels of H-D exchange were observed to be protein-dependent. Their analysis revealed these variations to be partly correlated to secondary structure. For each protein, a set of 6 spectra was measured with a systematic variation of the solvent H-D ratio and was subjected to factor analysis. The most significant component spectra for each protein, representing independent aspects of the spectral response to deuteration, were each subjected to a second factor analysis over the entire training set. Restricted multiple regression (RMR) analysis using the loadings of the principal components from 19 of these H-D analyses revealed an improvement in prediction accuracy compared with conventional bandshape-based analyses of FTIR data. Nearly a factor of 2 reduction in error for prediction of helix fractions was found using s1, the average spectral response for the H-D set. In some cases, significant error reduction for prediction of minor components was found using higher factors. Using the same analytical methods, prediction errors with this new deuteration-response-FTIR method were shown to be even better than those obtained by use of electronic circular dichroism (ECD) data for helix predictions and to be significantly lower for ECD-based sheet prediction, making these the best secondary structure predictions obtained with the RMR method. Tests of a limited variable selection scheme showed further improvements, consistent with previous results of this approach using ECD data. PMID- 10805521 TI - Coumarin-Ser-Asp-Lys-Pro-OH, a fluorescent substrate for determination of angiotensin-converting enzyme activity via high-performance liquid chromatography. AB - N-Acetyl-Ser-Asp-Lys-Pro-OH (AcSDKP-OH), a negative regulator of hematopoietic stem cell proliferation, is shown to be a physiological substrate of angiotensin I-converting enzyme (ACE), a zinc-dipeptidyl carboxypeptidase, involved in cardiovascular homeostasis. Recently, a study carried out on captopril-treated volunteers revealed that the kinetics of [3H]AcSDKP-OH hydrolysis in vitro in the plasma of donors correlates closely to the plasmatic ratio angiotensin II/angiotensin I, which characterized the conversion activity of ACE. This prompted us to design a fluorescent substrate, 2-[7-(dimethylamino)-2-oxo-2H chromen-4-yl]acetyl-SDKP-OH, or coumarin-SDKP-OH, which could be an alternative to the radiolabeled analogue used in that study, allowing an easier and more rapid determination of enzyme activity. We report here the synthesis and the determination of the kinetics constants of this fluorescent derivative compared with those of [3H]AcSDKP-OH with human plasma ACE (133 and 125 microM, respectively), which are in the same range as those of the physiological substrate angiotensin I. Furthermore, the hydrolysis of the fluorescent substrate shows the same sensitivity toward chloride concentration as the natural substrate, demonstrating its specificity for N-domain hydrolysis. This fluorescent derivative was used to develop a sensitive assay for the determination of ACE activity in human plasma. PMID- 10805522 TI - Quantification of long-chain aldehydes by gas chromatography coupled to mass spectrometry as a tool for simultaneous measurement of plasmalogens and their aldehydic breakdown products. AB - The cleavage of the specific vinyl ether linkage at the sn-1 position of plasmalogens leads to the formation of two products: the 1-lyso-2-acyl glycerophospholipid and a long-chain fatty aldehyde. Plasmalogens are measured by quantifying one of these two products. In this paper, we describe a rapid and sensitive procedure for measuring plasmalogens via quantification of long-chain fatty aldehydes. After lipid extraction, the sn-1 vinyl ether bond of plasmalogens is cleaved by acidic hydrolysis. The produced aldehydes are then derivatized with (pentafluorobenzyl)hydroxylamine hydrochloride and analyzed by gas chromatography/mass spectrometry in selected-ion mode. Plasmalogens are then indirectly quantified by subtracting the free aldehydes obtained without prior HCl treatment from the total aldehydes obtained after acidic hydrolysis. This method is applied to three rat brain areas selected for this study. Two of these are affected in neurodegenerative diseases (cerebral cortex and hippocampus) and one is rich in white matter (cerebellum). In comparison to other procedures, the advantages of this method are not only its usefulness in plasmalogen quantification but also the identification of aldehydic breakdown products. PMID- 10805523 TI - Streamlined F2-isoprostane analysis in plasma and urine with high-performance liquid chromatography and gas chromatography/mass spectroscopy. AB - F2-Isoprostanes in plasma and urine are generally determined by labor-intensive methods requiring sample purification by solid-phase extraction and thin-layer chromatography (TLC). A streamlined and more sensitive method for the measurement of esterified plasma F2-isoprostanes was developed by replacing these steps with high-performance liquid chromatography (HPLC) using an amino column with a hexane/2-propanol gradient. Pentafluorobenzyl esters of F2-isoprostanes were prepared and purified by HPLC, silylated, and then analyzed by gas chromatography (GC) with negative chemical ionization mass spectroscopy (NCI-MS). This method permits analysis with lower plasma volumes (100 microL) and greater sensitivity (to 10 pg; allowing detection to 50 pg/mL) than provided by other methods. Urinary F2-isoprostanes can also be efficiently quantified by this method, with 8 iso-PGF2alpha being identified as a major isomer. With this procedure, esterified plasma F2-isoprostanes were found to be 8.3-fold higher in an end-stage renal failure patient on hemodialysis and urinary 8-iso-PGF2alpha was 7.1-fold higher in a cigarette smoker than respective control subjects. This method, particularly the substitution of the TLC step common to other methods with HPLC, results in a more sensitive and reproducible assay. PMID- 10805524 TI - Determining glutathione and glutathione disulfide using the fluorescence probe o phthalaldehyde. AB - Because of the importance of glutathione (GSH) and glutathione disulfide (GSSG) in cellular signal transduction, gene regulation, redox regulation, and biochemical homeostasis, accurate determination of cellular glutathione levels is critical. Several procedures have been developed, but many suffer from overestimating GSSG or from cellular substances interfering or competing with GSH determination. Assays based on HPLC, with enzymatic reduction of GSSG by glutathione reductase and NADPH, appear to be valid but are limited in sample throughput and availability of equipment. The fluorescence probe o-phthalaldehyde (OPA, phthalic dicarboxaldehyde) reacts with GSH and has a high quantum yield, yet its use has been limited due to unidentified interfering and fluorescence quenching substances in liver. This paper describes assay conditions under which these limitations are avoided. By using a phosphate-buffered assay at lower pH, interference with nonspecific reactants is minimal. Since enzymatic reduction is not possible due to the reaction of OPA with NAD(P)H and other stronger reducing agents, leading to an overestimation of GSSG levels, dithionite was used to reduce GSSG. High sample throughput combined with sensitive (20-pmol limit of detection) and accurate determination of GSH and GSSG using OPA is achievable with any monochromatographic spectrofluorometer. Sample preparation and storage conditions are described that return the same levels of GSH and GSSG for at least 4 weeks. PMID- 10805525 TI - Quantification of bioactive acylethanolamides in rat plasma by electrospray mass spectrometry. AB - We developed a high-performance liquid chromatography/mass spectrometry (HPLC/MS) method for the identification and quantification of anandamide, an endogenous cannabinoid substance, and other fatty acid ethanolamides (AEs) in biological samples. Using a mobile-phase system of methanol/water and gradient elution, we achieved satisfactory resolution of all major AEs, including anandamide, palmitylethanolamide (PEA), and oleylethanolamide (OEA). Electrospray-generated quasi-molecular species were used as diagnostic ions and detected by selected ion monitoring (SIM). Synthetic deuterium-labeled AEs were used as internal standards, and quantification was carried out by isotope dilution. A linear correlation (r2 = 0.99) was observed in the calibration curves for standard AEs over the range 0-0.5 nmol. Detection limits between 0.1 and 0.3 pmol per sample and quantification limits between 0.5 and 1.2 pmol per sample were obtained. The method was applied to the quantification of anandamide, PEA, and OEA in plasma prepared from rat blood collected either by cardiac puncture or by decapitation. After cardiac puncture, AE levels were in the low-nanomolar range: anandamide, 3.1 +/- 0.6 pmol/ml; PEA, 9.4 +/- 1.6 pmol/ml; OEA, 9.2 +/- 1.8 pmol/ml (mean +/- SE, n = 9). By contrast, after decapitation AEs were dramatically elevated (anandamide, 144 +/- 13 pmol/ml; PEA, 255 +/- 55 pmol/ml; OEA, 175 +/- 48 pmol/ml). Thus, disruptive procedures of blood collection may result in gross overestimates in the concentrations of circulating AEs. PMID- 10805526 TI - Quantitative affinity chromatographic studies of mitochondrial cytochrome c binding to bacterial photosynthetic reaction center, reconstituted in liposome membranes and immobilized by detergent dialysis and avidin--biotin binding. AB - In order to study the affinity binding of c-type cytochromes to the photosynthetic reaction center (RC) by quantitative affinity chromatography (QAC), RC from Rhodobacter sphaeroides was reconstituted into liposomes composed of egg phosphatidylcholine (EPC) and 2 mol% of biotinyl phosphatidylethanolamine simultaneously as the liposomes were formed and immobilized in (strept)avidin coupled gel beads by rotary detergent dialysis. The immobilized amount was up to 80 nmol of RC and 33 micromol of lipid/g of moist gel in streptavidin-coupled Sephacryl S-1000 gel. By QAC frontal runs, retardation of mitochondrial cyt c on immobilized RC liposome columns was demonstrated. The dissociation constant for the RC-cyt c interaction was determined to be 0.20-0.57 microM. QAC studies also allowed evaluation of the orientation of reconstituted RC in immobilized liposomes by comparison of the total amount of cyt c binding sites with the amount of available binding sites obtained by QAC. It seems that the RC proteoliposomes immobilized in Sephacryl S-1000 gel exposed the cyt c binding sites on the outer surface of the liposomes due to effects of the gel network pore size and the resulting liposomal size. PMID- 10805527 TI - Single-nucleotide polymorphism analysis by pyrosequencing. AB - There is a growing demand for high-throughput methods for analysis of single nucleotide polymorphic (SNP) positions. Here, we have evaluated a novel sequencing approach, pyrosequencing, for such purposes. Pyrosequencing is a sequencing-by-synthesis method in which a cascade of enzymatic reactions yields detectable light, which is proportional to incorporated nucleotides. One feature of typing SNPs with pyrosequencing is that each allelic variant will give a unique sequence compared to the two other variants. These variants can easily be distinguished by a pattern recognition software. The software displays the allelic alternatives and allows for direct comparison with the pyrosequencing raw data. For optimal determination of SNPs, various protocols of nucleotide dispensing order were investigated. Here, we demonstrate that typing of SNPs can efficiently be performed by pyrosequencing using an automated system for parallel analysis of 96 samples in approximately 5 min, suitable for large-scale screening and typing of SNPs. PMID- 10805528 TI - Quantitative radioisotopic determination of histidine decarboxylase using high performance liquid chromatography. AB - We have developed a procedure for the accurate measurement of histidine decarboxylase in tissues expressing low levels of enzymatic activity. Briefly, histamine is enzymatically synthesized from [3H]-labeled histidine, followed by purification using high-performance liquid chromatography (HPLC) and quantitation by liquid scintillation counting. This method presents three advantages over previous techniques. First, prior to HPLC purification, excess precursor [3H]histidine is removed on an anion-exchange resin. Second, purification by HPLC is considerably more selective than that of classical cation-exchange gravity columns or organic solvent extractions. Finally, the accuracy of this method is improved by including non-radiolabeled histamine as internal standard, which is quantified by ultraviolet detection. This simple procedure allows highly sensitive and accurate determinations of histamine synthesis. PMID- 10805529 TI - Development of an ultralow-light-level luminescence image analysis system for dynamic measurements of transcriptional activity in living and migrating cells. AB - We have developed an approach to study in single living epithelial cells both cell migration and transcriptional activation, which was evidenced by the detection of luminescence emission from cells transfected with luciferase reporter vectors. The image acquisition chain consists of an epifluorescence inverted microscope, connected to an ultralow-light-level photon-counting camera and an image-acquisition card associated to specialized image analysis software running on a PC computer. Using a simple method based on a thin calibrated light source, the image acquisition chain has been optimized following comparisons of the performance of microscopy objectives and photon-counting cameras designed to observe luminescence. This setup allows us to measure by image analysis the luminescent light emitted by individual cells stably expressing a luciferase reporter vector. The sensitivity of the camera was adjusted to a high value, which required the use of a segmentation algorithm to eliminate the background noise. Following mathematical morphology treatments, kinetic changes of luminescent sources were analyzed and then correlated with the distance and speed of migration. Our results highlight the usefulness of our image acquisition chain and mathematical morphology software to quantify the kinetics of luminescence changes in migrating cells. PMID- 10805530 TI - Time-resolved spectral observations of cadmium-enriched cadmium sulfide nanoparticles and the effects of DNA oligomer binding. AB - We measured the steady-state and time-resolved fluorescence spectral properties of cadmium-enriched nanoparticles (CdS-Cd2+). These particles displayed two emission maxima, at 460 and 580 nm. The emission spectra were independent of excitation wavelength. Surprisingly, the intensity decays were strongly dependent on the observation wavelength, with longer decay times being observed at longer wavelengths. The mean lifetime increased from 150 to 370 ns as the emission wavelength was increased from 460 to 650 nm. The wavelength-dependent lifetimes were used to construct the time-resolved emission spectra, which showed a growth of the long-wavelength emission at longer times, and decay-associated spectra, which showed the longer wavelength emission associated with the longer decay time. These nanoparticles displayed anisotropy values as high as 0.35, depending on the excitation and emission wavelengths. Such high anisotropies are unexpected for presumably spherical nanoparticles. The anisotropy decayed with two correlation times near 5 and 370 ns, with the larger value probably due to overall rotational diffusion of the nanoparticles. Addition of a 32-base pair oligomer selectively quenched the 460-nm emission, with less quenching being observed at longer wavelengths. The time-resolved intensity decays were minimally affected by the DNA, suggesting a static quenching mechanism. The wavelength selected quenching shown by the nanoparticles may make them useful for DNA analysis. PMID- 10805531 TI - A homogeneous cell-based assay to identify N-linked carbohydrate processing inhibitors. AB - Malignant transformation is accompanied by altered cell surface glycosylation. N Linked oligosaccharides carrying beta1-6GlcNAc branches are associated with tumor invasion and metastasis. Therefore, compounds that can enter cells and block biosynthesis of beta1-6GlcNAc-branched glycans without overt cytotoxicity are potential anticancer agents. We have developed a homogeneous cell-based assay for detection of such compounds. The method enables identification of agents that block beta1-6GlcNAc-branched glycan expression after incubation for 16-20 h with MDAY-D2 tumor cells, thereby protecting the cells from the subsequent addition of leukoagglutinin, a cytotoxic plant lectin. We observed that MDAY-D2 cell number is directly proportional to the level of endogenous alkaline phosphatase activity measured spectrophotometrically in cultures after the addition of substrate. The alkaline phosphatase assay was capable of detecting as few as 1,500 cells. The assay was readily adapted for high-throughput screening as reagent costs are low and no cell harvesting and washing steps are required. Under high-throughput operating conditions, the coefficient of variation for controls was found to be 4.2%. The results suggest that measurement of alkaline phosphatase in this cell assay format may be adapted for wider applications in high-throughput screenings for compounds that relieve cells from other growth inhibitors. PMID- 10805532 TI - Comparison between different strategies of covalent attachment of DNA to glass surfaces to build DNA microarrays. AB - DNA microarray is a powerful tool allowing simultaneous detection of many different target molecules present in a sample. The efficiency of the array depends mainly on the sequence of the capture probes and the way they are attached to the support. The coupling procedure must be quick, covalent, and reproducible in order to be compatible with automatic spotting devices dispensing tiny drops of liquids on the surface. We compared several coupling strategies currently used to covalently graft DNA onto a glass surface. The results indicate that fixation of aminated DNA to an aldehyde-modified surface is a choice method to build DNA microarrays. Both the coupling procedure and the hybridization efficiency have been optimized. The detection limit of human cytomegalovirus target DNA amplicons on such DNA microarrays has been estimated to be 0.01 nM by fluorescent detection. PMID- 10805533 TI - Immunoliposome sandwich assay for the detection of Escherichia coli O157:H7. AB - We describe the development of a field-portable colorimetric immunoassay for the detection of Escherichia coli O157:H7, using antibody-directed liposomes (immunoliposomes) encapsulating dye as an analytical reagent. Antibodies (anti-E. coli O157:H7) thiolated by 2-iminothiolane were coupled to malemide-tagged liposomes encapsulating the marker dye, sulforhodamine B. Transmission electron microscopy showed that the immunoliposomes bound only to the serotype without any cross-reactivity with tested negative controls. A wicking reagent containing immunoliposomes and the test sample and a plastic-backed nitrocellulose strip with a measurement zone were used in a sandwich (noncompetitive) assay format. During the capillary migration of the wicking reagent, E. coli, with surface bound immunoliposomes, was captured at the measurement zone on which antibodies to E. coli O157:H7 were immobilized. The color density of the measurement zone was directly proportional to the amount of E. coli O157:H7 in the sample. The detection limit of the current assay with pure cultures of the serotype was ca. 10(4) colony-forming units (CFU)/mL. The assay, which does not need washing and incubation steps, can be completed in 8 min. These results demonstrate the feasibility of using dye-encapsulating immunoliposomes in microporous membranes for the rapid detection of molecules with multivalent antigenic sites. PMID- 10805534 TI - Two continuous spectrophotometric assays for methionine aminopeptidase. AB - Two spectrophotometric assays have been developed for methionine aminopeptidases (MetAPs). The first method employs a thioester substrate which, upon enzymatic removal of the N-terminal methionine, generates a free thiol group. The released thiol is quantitated using Ellman's reagent. The MetAP reaction is conveniently monitored on a UV-VIS spectrophotometer in a continuous fashion, with the addition of an excess of Ellman's reagent into the assay reaction. Two tripeptide analogues were synthesized and found to be excellent substrates of both Escherichia coli MetAP and human MetAP2 (k(cat)/K(M) = 2.8 x 10(5) M(-1) s(-1) for the most reactive substrate). In the second assay method, the MetAP reaction is coupled to a prolyl aminopeptidase reaction using Met-Pro-p-nitroanilide as substrate. MetAP-catalyzed cleavage of the N-terminal methionine produces prolyl p-nitroanilide, which is rapidly hydrolyzed by the prolyl aminopeptidase from Bacillus coagulans to release a chromogenic product, p-nitroaniline. This allows the MetAP reaction to be continuously monitored at 405 nm on a UV-VIS spectrophotometer. The assays have been applied to determine the pH optima and kinetic constants for the E. coli and human MetAPs as well as to screen MetAP inhibitors. These results demonstrate that the current assays are convenient, rapid, and sensitive methods for kinetic studies of MetAPs and effective tools for screening MetAP inhibitors. PMID- 10805535 TI - Molecular beacons: a real-time polymerase chain reaction assay for detecting Salmonella. AB - Molecular beacons are oligonucleotide probes that become fluorescent upon hybridization. We developed a real-time PCR assay to detect the presence of Salmonella species using these fluorogenic reporter molecules. A 122-base-pair section of the himA was used as the amplification target. Molecular beacons were designed to recognize a 16-base-pair region on the amplicon. As few as 2 colony forming unit (CFU) per PCR reaction could be detected. We also demonstrated the ability of the molecular beacons to discriminate between amplicons obtained from similar species such as Escherichia coli and Citrobacter freundii in real-time PCR assays. These assays could be carried out entirely in sealed PCR tubes, enabling fast and direct detection of Salmonella in a semiautomated format. PMID- 10805536 TI - A facile enzymatic synthesis of uridine diphospho-[14C]galacturonic acid. AB - Galacturonic acid (GalA) is a major component of plant cell-wall-derived pectins. It can be also found in the cell-surface polysaccharides of different microorganisms, including several symbiotic and pathogenic bacteria. Uridine diphosphogalacturonic acid (UDP-GalA) is a likely donor for GalA during the biosynthesis of these polysaccharides. A highly efficient, yet simple, method is presented for generating and purifying UDP-[14C]GalA. Commercially available UDP [14C]-galactose was quantitatively oxidized (>95% conversion) to UDP-[14C]GalA in the presence of high levels of galactose oxidase and catalase, at prolonged incubation times. Following this one-step enzymatic oxidation, UDP-[14C]GalA was purified using a polyethyleneimine cellulose column with a single-step 1 M NaCl elution. The authenticity of the purified UDP-[14C]GalA was verified by its relative mobility on thin-layer chromatograms, analysis of its chemical hydrolysis products, and 1H NMR spectroscopy. Our yield of >90% is much higher than by previously described methods. The method may serve as a prototype for the preparation of other radiolabeled uronic acids and their nucleotide derivatives. PMID- 10805537 TI - A method of immobilization on the solid support of complex and simple enzymes retaining their activity. AB - Immobilization of native proteins, retaining their activity, on the solid support is often crucial for a variety of biochemical assays involving protein-protein interactions. In this study we describe a technique which allows binding of both complex (protein kinase CK2) and simple (calf intestine alkaline phosphatase, CIP) enzymes to the solid support without denaturization of the proteins. This method is based on the covalent cross-linking of the enzymes to the bifunctional resin, containing the secondary amino and thiol groups, in a coupling reaction with the imidoester dimethyl pimelimidate hydrochloride. Both enzymes in their bound form were active in the specific biochemical assays. We also found that the CK2 and CIP resins did not change their activity for at least 3 months, and the quality of these resins were not affected by high salts or reducing agents. Thus, this method can be recommended for general use to generate active enzymes coupled to the solid support. PMID- 10805538 TI - Monitoring of isotope substitution in cyanobacteria by matrix-assisted laser desorption ionization--time-of-flight mass spectrometry. PMID- 10805539 TI - Mapping 3' termini of mRNA on DNA templates with Taq polymerase 3'-end-labeled probes. PMID- 10805540 TI - Measurement of apoptosis by the TUNEL method using scintillating microplates. PMID- 10805541 TI - Elimination of artifactual bands from polyacrylamide gels. PMID- 10805542 TI - Solid-phase labeling with a fluorescent reagent to fingerprint nonradioactive proteins. PMID- 10805543 TI - Pepsin inactivation of deoxyribonuclease I. PMID- 10805544 TI - Liver anatomy: portal (and suprahepatic) or biliary segmentation. AB - BACKGROUND/AIMS: In liver anatomy and surgery, is portal and hepatic vein segmentation (French segmentation) to be preferred over arteriobiliary segmentation (Healey and Schroy, North American segmentation)? METHODS: Several embryological arguments and an analysis of anatomical data from a personal collection of 110 vasculobiliary casts were made. RESULTS: Embryological arguments: Portal vein branching appears first, arteriobiliary branching secondly follows the portal vein distribution. Segment II (the left lateral sector) is the development of the right lateral embryological lobe. The umbilical vein enters the left portion of the middle embryological lobe, forming segment IV on the right and segment III on the left: this is the left paramedian sector. So the left portal fissure (between left and middle lobes) transversally crosses the classical left lobe, which is not a portal unit. Segment VI is a late secondary prominence of segment VII, reaching the anterior margin of the liver only in man. Anatomical arguments: hepatic vein segmentation must be added to portal segmentation; the academic left lobe is the left hepatic vein sector, and the left hepatic fissure separates the classical right and left lobes. Portal vein segmentation must be preferred: portal vein duplication of branches of first order occurs only in 23.5% of the cases, while arteriobiliary duplication of first-order branches is noted in 50% of the livers, portal segmentation being much simpler. CONCLUSIONS: Portal and hepatic vein segmentation seems to be much more accurate. PMID- 10805545 TI - Proximal gastrectomy and gastric tube reconstruction for early cancer of the gastric cardia. AB - BACKGROUND: Advances in diagnostics and techniques have increased the number of detections of early cancer of the gastric cardia. The aim of this report is to describe a new technique of proximal gastrectomy and gastric tube reconstruction using an autosuture stapler. METHODS: At operation, the upper part of the stomach was fully mobilized, and the abdominal esophagus was transected. The stomach was cut between the points of the distal three fourths of the lesser curvature and a half of the greater curvature, and a gastric tube measuring 20 cm in length and 4 cm in width was made. A circular stapler was inserted through a stapled line, and direct anastomosis between the esophagus and gastric tube was completed. RESULTS: When compared with the patients who underwent proximal gastrectomy and jejunal interposition, those who underwent gastric tube reconstruction showed a significantly shorter operation time. All 3 patients who underwent proximal gastrectomy and gastric tube reconstruction recovered uneventfully and took foods satisfactorily. Good passage through the gastric tube was recognized on roentgenography, and reflux esophagitis or stomal ulcer was not found on fiberscopy. CONCLUSIONS: We believe the procedure is technically simple and safe, and provides good postoperative results in patients with early cancer of the gastric cardia. PMID- 10805547 TI - Correlation of tensile strength with bursting pressure in the evaluation of intestinal anastomosis. AB - BACKGROUND: Although bursting pressure and tensile strength have long been measured to evaluate anastomotic techniques, it has yet to be clarified whether or not they are correlated, what implications they have, and which should be used as a gold standard. MATERIAL AND METHODS: Using an experimental model to estimate pressure and tension in the same colonic anastomosis, the following variables were measured in 48 rats between days 0 and 14: bursting pressure (BP); minimal tensile strength (MITS) necessary to break a part of the anastomosis, and maximal tensile strength (MATS) needed to disrupt the whole anastomosis. Also, circulatory wall tension (CWT) was derived from BP and the anastomotic circumference (AC), and longitudinal wall tension (LWT) from MITS and AC. These variables were compared using correlation and regression analysis. RESULTS: During the lag phase (days < or = 4) there was poor correlation between pressure related and tension-related variables whereas highly significant correlations were noted in the subsequent fibroplastic phase (day > or = 5). It was shown by regression lines that positive MITS and MATS were expected when BP was zero. CONCLUSION: Contrary to the previous assumption, no correlation was found between BP and tensile strength in the critical postoperative period. Based on our present and previous studies, measurement of MITS is recommended to evaluate the healing of colonic anastomosis. PMID- 10805546 TI - Systemic and cell-mediated immune response after laparoscopic and open cholecystectomy in patients with chronic liver disease. A randomized, prospective study. AB - BACKGROUND: As impaired immune function observed in cirrhotic patients is known to increase the risk of postoperative complications, the immunological response to surgery was investigated. METHODS: Twenty-eight patients with postnecrotic liver cirrhosis or chronic hepatitis C and symptomatic gallstone disease were randomly allocated to laparoscopic (LC) or open cholecystectomy (OC). Changes in concentrations of cytokines (TNF-alpha, IL-1beta, IL-6, IL-8 and IL-10) were followed and the effect of surgical trauma on the distribution of lymphocyte subpopulations (CD3, CD4, CD8, CD16 and CD19) and NK cell cytotoxicity were measured. RESULTS: After OC a decrease in circulating CD3 (p < 0.05) and CD4 (p < 0.05) and an increase in CD19 (p < 0.05) cells were detected in contrast to LC after which only CD16 cells decreased (p = 0.05). The number of CD3 cells was higher after LC than after OC (p < 0.01), whereas the number of CD19 cells was higher after OC than after LC (p < 0.01). NK cell cytotoxicity was reduced after LC (p < 0.05). In cirrhotic patients circulating cytokines were unaffected by OC, whereas TNF-alpha (p < 0.05) and IL-1beta (p < 0.05) were reduced after LC. In chronic hepatitis IL-1beta decreased after OC (p = 0.05) and IL-10 was significantly higher after LC than following OC (p < 0.05). CONCLUSION: The immune response is less pronounced after a laparoscopic procedure compared to a conventional approach in patients with chronic liver disease. PMID- 10805548 TI - Protein C activation during the initial phase of experimental acute pancreatitis in the rabbit. AB - BACKGROUND: Disturbances of coagulation and fibrinolysis are well-known systemic effects of acute necrotising pancreatitis (ANP). The purpose of this experimental study was to evaluate the initial events in the haemostatic activation during ANP in an animal model with relevance to the human situation. METHODS: ANP was introduced in 7 rabbits by infusion of chenodeoxycholic acid in the pancreatic duct. Seven rabbits served as sham-operated controls. Serial measurements of coagulation variables (prothrombin time, activated partial thromboplastin time, FVII activity, fibrinogen, tissue factor activity), anticoagulant proteins (protein C, antithrombin) and fibrinolytic factors (tissue plasminogen activator, plasminogen activator inhibitor-1) were performed for 5 h. RESULTS: ANP was confirmed by elevated serum amylase, development of ascites, and histological changes of the pancreas. A moderate activation of the coagulation system was found in both study groups. A significant decrease in protein C concentration from 1 h after the induction of ANP was found, whereas the response of antithrombin and the inhibition of the fibrinolytic system were similar in the 2 study groups. Microthrombosis of the lungs or kidneys was found in 2 rabbits with ANP. CONCLUSION: An immediate activation of protein C is a specific characteristic of the haemostatic activation in ANP in rabbits. This activation has not been described previously and the possible therapeutic implications ought to be studied. PMID- 10805549 TI - Beta-cell function of the pancreas after necrotizing pancreatitis. AB - AIM: To investigate the late sequellae of necrotizing pancreatitis on the endocrine function of the pancreas. PATIENTS AND METHODS: Twenty patients, 15 men (mean +/- SEM age 52.2+/-2.6 years and BMI 26.8+/-0.8 kg/m2) and 5 women (age 51.0+/-7.6 years and BMI 26.7+/-0.8 kg/m2) were submitted to a glucagon stimulation test 63 (range 8-136) months after an attack of pancreatitis. All nondiabetic patients (n = 15) were also submitted to an oral glucose tolerance test. For comparison, 16 healthy volunteers, 8 men (age 56.0+/-0.9 years and BMI 26.3+/-0.4 kg/m2) and 8 women (age 50.5+/-1.0 years and BMI 28.2+/-0.6 kg/m2), were also studied. RESULTS: Five patients (25%) had diabetes mellitus and needed insulin treatment, 6 patients (30%) had an impaired glucose tolerance (IGT). Nondiabetic patients (IGT included) had a significantly higher basal insulin level (15.8+/-1.9 vs. 10.9 +/-2.2 mU/l, p < 0.05) and a lower glucose/insulin ratio (p < 0.05) compared with controls. The serum concentrations of insulin and C peptide, after stimulation with glucagon, calculated as peak value, maximal increment and as area under the curve were not significantly different in the nondiabetic patients compared to controls. The subgroup of IGT patients had a significantly higher basal C peptide (p < 0.05) and a reduced maximal increment (p < 0.05). CONCLUSIONS: After nonresectional therapy for necrotizing pancreatitis, there is a high prevalence of disturbances in glucose metabolism. Patients with IGT have signs of both loss of beta-cell function and insulin resistance. PMID- 10805550 TI - Internal drainage of infected pancreatic pseudocysts: safe or sorry? AB - BACKGROUND: External drainage is the traditional surgical therapy for infected pancreatic pseudocyst, although associated with high morbidity and mortality rates. In this study it was determined whether internal drainage is feasible with acceptable postoperative morbidity and recurrence rates. METHODS: A retrospective comparison was made of the outcome of internal versus external drainage of infected pseudocysts in 15 patients. All patients were known to have a (sterile) pseudocyst and presented with symptoms suggestive of infection of the cyst, proven by positive cultures and Gram staining. RESULTS: Internal drainage was performed in 8 and external drainage in 7 patients. Patient characteristics appeared comparable, as was the time of sterile cyst presence before infection occurred (6 and 9 weeks, respectively). No major complications occurred, although hospital stay was prolonged after external drainage due to development of pancreaticocutaneous fistulas along the drain tract in 4 patients. Enteric microorganisms were cultured in 11 patients, of whom 10 had undergone ERCP just prior to infection. During follow-up no pseudocyst recurred. CONCLUSION: Surgical internal drainage of an infected pseudocyst is safe and effective and, in selected patients, is first-choice treatment. ERCP seems to play an important role in the secondary infection of pseudocysts. PMID- 10805551 TI - A surgical treatment of infected pancreatic necrosis: retroperitoneal laparotomy. AB - BACKGROUND/AIMS: Due to the anatomical location of the pancreas, sufficient drainage of a pancreatic necrosis by laparotomy may be unsatisfactory. METHODS: CT and helical CT have provided extremely useful information on the surgical treatment of necrotic pancreatitis. The retroperitoneal approach (RPA) allows direct and complete removal of necrotic tissues. RESULTS: RPA was used to treat 8 patients with infected pancreatic necrosis. Excision of necrotic tissues was effective and could minimize the complications often associated with laparotomy such as bleeding and intestinal injuries. CONCLUSION: By CT and helical CT, three dimensional images of pancreatic necrosis are obtained. These investigations have greatly facilitated RPA, which has advantages over laparotomy in the treatment of infected pancreatic necrosis. PMID- 10805552 TI - Wound complications after major gastrointestinal operations. The surgeon as a risk factor. AB - BACKGROUND: Wound complications occur quite often after abdominal operations. Numerous studies have been performed in the last decades focusing on closure methods, incisions and suture materials. However, the most important factor, the individual surgeon, has hardly been taken into account in these studies. METHODS: This study presents results from a prospective randomised study on abdominal wall closure focusing on the results of the individual surgeon. RESULTS/CONCLUSIONS: We found no differences in the complication rate between different suture materials or between continuous and interrupted closure techniques. There are marked individual differences in complication rates between surgeons. Regular audit with feedback to individual surgeons is an important instrument for quality improvement. PMID- 10805553 TI - Botulinum toxin for the treatment of anal fissure. AB - BACKGROUND: The classic treatment for uncomplicated anal fissure is surgical sphincterotomy, i.e. cutting of the internal anal sphincter, thus eliminating spasm of this muscle and breaking the vicious circle of pain, spasm and inflammation. Recently, however, botulinum toxin has become available for the treatment of muscular dystonias, and thus for anal fissure. In the present study, we investigated the effectiveness of treatment with botulinum toxin in 76 patients with uncomplicated anal fissure. MATERIAL AND METHOD: The 76 patients received an injection of 40 U of botulinum toxin on each side of the fissure. Response was monitored 7, 30 and 90 days later. All patients who did not show clear improvement after 30 days received a second dose of 40 U on each side. RESULTS: After 90 days, 51 patients (67%) showed complete recovery, 19 patients (25%) substantial improvement though not complete recovery, and 6 patients (8%) no significant improvement. Transitory gas incontinence was reported by 2 patients (2.6%), and 1 patient presented hemorrhoidal thrombosis. DISCUSSION: Botulinum toxin enables chemical denervation of the internal sphincter, facilitating healing of the anal fissure. Its principal advantages with respect to surgical sphincterotomy are the absence of the general risks of surgery, and reduced incidence of incontinence, which even if it occurs tends to be transitory. The technique does not require hospitalization and is well tolerated. It appears suitable for the initial treatment of uncomplicated anal fissure, reserving surgical treatment for those cases which fail to response adequately. PMID- 10805554 TI - Laparoscopic cholecystectomy in two patients with symptomatic cholelithiasis and situs inversus totalis. AB - BACKGROUND: Situs inversus viscerum is a rare condition with a genetic predisposition. We report 2 patients with situs inversus totalis and symptomatic cholelithiasis successfully treated via laparoscopic cholecystectomy. PATIENTS AND METHODS: The first patient was a 61-year-old female presenting with pain in the left upper quadrant associated with fever, chills, nausea and vomiting. The abdomen was tender with guarding and rebounding pain in the same region. Abdominal ultrasound and CT scan confirmed the diagnosis of gallstones as well as situs inversus with the liver and gallbladder on the left side and the spleen on the right. The second patient was a 37-year-old male with known situs inversus who presented with biliary colic due to cholelithiasis. In both patients cholecystectomy was performed laparoscopically in a reverse fashion. RESULTS: Laparoscopic cholecystectomy was carried out successfully despite the reversed anatomic relationships and both patients made a smooth recovery. CONCLUSION: Cholelithiasis occurring with situs inversus totalis is rare and may present a diagnostic problem. Laparoscopic cholecystectomy can be safely and effectively applied in the setting of situs inversus, although attention must be paid to the details of left-right reversal. PMID- 10805555 TI - Small bowel ischemia following laparoscopic cholecystectomy. AB - BACKGROUND: Among a myriad of physiological adverse affects of pneumoperitoneum associated intra-abdominal hypertension, compromise of the mesenteric circulation is well documented. METHODS: After experiencing a case of fatal small bowel ischemia in the aftermath of laparoscopic cholecystectomy, the literature was reviewed. RESULTS: A Medline and Index Medicus search revealed at least 6 cases of small bowel ischemia following laparoscopic cholecystectomy. CONCLUSIONS: Mesenteric ischemia should be considered in the differential diagnosis of patients developing nonspecific abdominal symptoms after laparoscopic procedures. PMID- 10805556 TI - Bouveret's syndrome complicated by acute pancreatitis. AB - BACKGROUND/AIM: This study evaluated a case of Bouveret's syndrome due to a cholecystoduodenal fistula and gallstone obstruction of the duodenum, complicated by acute pancreatitis and cholecystitis. METHODS: The presenting features, special investigations, radiological findings, operative and endoscopic procedures were reviewed. RESULTS: Symptoms persisted after laparotomy and removal of a gallstone in the duodenum. Intra-operative endoscopy identified a second previously undetected stone impacted in the distal duodenum. CONCLUSION: The importance of excluding more than one stone causing Bouveret's syndrome is emphasized. PMID- 10805557 TI - Successful nonoperative management of bleeding hepatic artery pseudoaneurysm following pancreaticoduodenectomy. AB - BACKGROUND/AIMS: To report the management of a hepatic artery pseudoaneurysm presenting 35 days following a Whipple procedure. METHODS: The case study of a patient with a bleeding pseudoaneurysm is presented. RESULTS: Computed tomography demonstrated a pseudoaneurysm which was successfully embolized. CONCLUSIONS: Acute gastrointestinal bleeding from a pseudoaneurysm in the hepatic artery following Whipple procedure can be successfully managed with transcatheter embolization. PMID- 10805558 TI - Multidisciplinary approach to palliation of obstructive jaundice caused by a central hepatocellular carcinoma. AB - BACKGROUND/AIMS: Obstructive jaundice due to intraductal tumour growth is a rare symptom in association with hepatocellular carcinoma (HCC). METHODS: We report a 65-year-old white male who was admitted to our department with a 2-week history of progressive jaundice. At laparotomy, the liver showed advanced cirrhosis due to long-standing biliary obstruction. Cholangiography confirmed total obstruction of the main bifurcation of the hepatic duct by intraductal tumour growth. Combination treatment with surgical segment III drainage, transcatheter arterial embolization and radioembolization with yttrium-90 resin particles and endoscopic stenting was performed. This form of treatment has never been reported before. RESULTS: With these combined procedures, relief of jaundice and a survival time of 32 months could be achieved. CONCLUSION: The combination of palliative methods may relieve jaundice, ensure a good quality of life and possibly prolong survival in patients with mechanical tumour obstruction of the biliary tree by HCC. PMID- 10805559 TI - Ongoing clinical trials. PMID- 10805560 TI - Acidophiles in bioreactor mineral processing. AB - Mineral processing in bioreactors has become established in several countries during the past decade with industrial application of iron- and sulfur-oxidizing bacteria to release occluded gold from mineral sulfides. Cobalt extraction in bioreactors has also been commercialized, and development of high-temperature biooxidation of copper sulfides has reached pilot-plant scale. A variety of potentially useful mineral sulfide-oxidizing thermophiles have been recognized, but the most active strains have not been fully characterized. PMID- 10805561 TI - Heavy metal-resistant bacteria as extremophiles: molecular physiology and biotechnological use of Ralstonia sp. CH34. AB - In contrast to thermophilic or psychrophilic organisms, heavy metal-resistant bacteria do not supply enzymes that are active under harsh conditions, but are themselves tools for the evaluation and remediation of heavy metal-contaminated environments. Ralstonia sp. CH34 is a gram-negative bacterium with a remarkable set of resistance determinants, allowing this bacterium to live in extreme environments that are heavily contaminated with toxic metal ions. These heavy metal ions are mostly detoxified by inducible ion efflux systems that reduce the intracellular concentration of a given ion by active export. Because all metal resistance determinants in this bacterium are inducible, their regulatory systems can be used to develop biosensors that measure the biologically important concentrations of heavy metals in an environment. Resistance based on metal ion efflux detoxifies only the cytoplasm of the respective cell. Therefore, this resistance mechanism cannot be used directly to develop biotechnological procedures; however, metal ion efflux can protect a cell in a metal-contaminated environment. Thus, the cell can be enabled to mediate biochemical reactions such as precipitation of heavy metals with the carbon dioxide produced during growth or degradation of xenobiotics. PMID- 10805562 TI - Toward a molecular understanding of cold activity of enzymes from psychrophiles. AB - Despite the fact that a much greater proportion of the earth environment is cold rather than hot, much less is known about psychrophilic, cold-adapted microorganisms compared with thermophiles living at high temperatures. In particular, investigation of the molecular basis of cold-active enzymes from psychrophiles has only recently received concerted research attention, in measure as a result of the EC-funded project COLDZYME. This research effort has been stimulated by the realization that such cold-active enzymes offer novel opportunities for biotechnological exploitation. Only very recently has the first cold-active enzyme, alpha-amylase, been crystallized, and this success was followed rapidly by others. This effort has facilitated a direct approach to solving the three-dimensional structure of cold-active enzymes to complement the gene homology modeling that had been performed previously. Recently studies have highlighted how different adaptations are used by different enzymes to achieve conformational flexibility at low temperatures, and how such adaptations are not necessarily the opposite of those that confer thermostability to proteins in thermophilic counterparts. This review also highlights initial successes in engineering genetically improved thermal stability in cold-active enzymes to give improved catalysts for low-temperature biotechnology. PMID- 10805563 TI - Halophilic adaptation of enzymes. AB - It is now clear that the understanding of halophilic adaptation at a molecular level requires a strategy of complementary experiments, combining molecular biology, biochemistry, and cellular approaches with physical chemistry and thermodynamics. In this review, after a discussion of the definition and composition of halophilic enzymes, the effects of salt on their activity, solubility, and stability are reviewed. We then describe how thermodynamic observations, such as parameters pertaining to solvent-protein interactions or enzyme-unfolding kinetics, depend strongly on solvent composition and reveal the important role played by water and ion binding to halophilic proteins. The three high-resolution crystal structures now available for halophilic proteins are analyzed in terms of haloadaptation, and finally cellular response to salt stress is discussed briefly. PMID- 10805564 TI - Analysis of the genome of an alkaliphilic Bacillus strain from an industrial point of view. AB - Bacillus species and other microbes with pH optima for growth higher than pH 9 are defined as alkaliphiles. A large number of alkaliphilic Bacillus strains producing useful enzymes, have been isolated from various environments. Some of these enzymes, such as proteases and cellulases from alkaliphilic Bacillus strains, have been commercialized and have brought great advantages to industry and domestic life. To support further development of the enzyme industry, we initiated analysis of the genome of Bacillus halodurans C-125, which is 4.25Mb in size, and constructed a physical and genetic map for comparison with the Bacillus subtilis chromosome. Systematic sequencing of the whole genome of Bacillus halodurans C-125 has been automated since the beginning of May 1998, and sequencing of 98% of the whole genome has been done so far. Through genome analysis, it became apparent that the genome organization of alkaliphilic Bacillus halodurans C-125 is totally different from that of B. subtilis orthologues. PMID- 10805565 TI - A novel bacteriorhodopsin-like protein from Haloarcula japonica strain TR-1: gene cloning, sequencing, and transcript analysis. AB - The gene encoding a novel bacteriorhodopsin (bR)-like protein from Haloarcula japonica strain TR-1 was cloned and sequenced. The nucleotide sequence of the gene contained an open reading frame that corresponded to a protein of 250 amino acids. The deduced amino acid sequence of Ha. japonica bR-like protein exhibited the highest homology to those of cruxrhodopsins (cRs) produced by members of the genus Haloarcula, suggesting that the bR-like protein of Ha. japonica belonged to the cR subfamily. The hydropathy analysis of Ha. japonica bR-like protein (cR) revealed that the Ha. japonica cR had a transmembrane heptahelical structure similar to that of bR. Furthermore, transcription of the cR gene in Ha. japonica was confirmed by the reverse transcription-polymerase chain reaction method. PMID- 10805566 TI - Molecular cloning, sequency, and expression of the heat-labile uracil-DNA glycosylase from a marine psychrophilic bacterium, strain BMTU3346. AB - The gene encoding a heat-labile uracil-DNA glycosylase (UDG) from a psychrophilic, gram-positive marine strain (BMTU3346) has been cloned, sequenced, and expressed in Escherichia coli. The UDG is a cold-active enzyme with an apparent temperature optimum of 35 degrees C and a half-life of 2min at 40 degrees C. The amino acid sequence shows an identity of 39.1%-46.2% to UDGs from mesophilic bacteria. The primary structure was examined for features that could be related to the thermolability of the enzyme. The amino acid sequence of the heat-labile UDG shows 22 differences with respect to the consensus sequence derived from bacterial UDGs. Features previously recognized in cold-active enzymes such as extended surface loops or a decrease in the number of arginine residues or proline residues in loops were not observed. Because dominant features that could be related to the thermolability of the UDG from BMTU3346 cannot be identified, more subtle modifications of the conformation seem to be responsible for its thermolability. PMID- 10805567 TI - Structural analysis of the elongation factor G protein from the low-temperature adapted bacterium Arthrobacter globiformis SI55. AB - The first structural analysis of elongation factor G (EF-G) from a cold-adapted bacterium is presented. EF-G is an essential protein involved in the elongation process during protein synthesis and is therefore thought to play a crucial role in the low-temperature adaptation of cold-adapted microorganisms. To define its importance, the EF-G gene (fus) from the psychrotolerant bacterium Arthrobacter globiformis SI55 was cloned and sequenced. The deduced primary structure of the elongation factor is composed of 700 amino acids with a predicted molecular mass of 77.4 kDa. A three-dimensional model of the protein was constructed based on the known crystal structures of structurally homologous proteins. Structural features that might potentially be important for activity and flexibility at low temperature were deduced by comparisons with models of the EF-G proteins from the closely related mesophiles Micrococcus luteus and Mycobacterium tuberculosis. These features include a loss in the number of salt bridges in intradomain and interdomain positions, increased solvent interactions mediated by greater charge and polarity on domain surfaces, loop insertions, loss of proline residues in loop structures, and an increase of hydrophobicity in core regions. Specific changes have also been identified in the catalytic domain (G domain) and sites of potential ribosome interaction, which may directly affect guanosine triphosphate (GTP) hydrolysis and elongation rates at low temperature. PMID- 10805568 TI - TMC-95A, B, C, and D, novel proteasome inhibitors produced by Apiospora montagnei Sacc. TC 1093. Taxonomy, production, isolation, and biological activities. AB - In our course of screening for novel proteasome inhibitors, TMC-95A and its diastereomers, TMC-95B to D, were isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093. TMC-95A inhibited the chymotrypsin-like (ChT L), trypsin-like (T-L), and peptidylglutamyl-peptide hydrolyzing (PGPH) activities of 20S proteasome with IC50 values of 5.4nM, 200nM, and 60nM, respectively. TMC-95B inhibited these activities to the same extent as TMC-95A, while the inhibitory activities of TMC-95C and D were 20 to 150 times weaker than that of TMC-95A and B. TMC-95A did not inhibit m-calpain, cathepsin L, and trypsin at 30 microM, suggesting its high selectivity for proteasome. Taxonomy of the producing strain is also described. PMID- 10805569 TI - CD45 tyrosine phosphatase inhibitory components from Aspergillus niger. AB - Two inhibitors of CD45 tyrosine phosphatase, dihydrocarolic acid (1) and penitricin D (2), were isolated from a fermentation broth of the fungus Aspergillus niger and purified by HSCCC (high speed countercurrent chromatography) followed by HPLC. The structures were determined by NMR. The inhibitory activities of both compounds were specific to tyrosine phosphatases. PMID- 10805570 TI - Curtisians A-D, new free radical scavengers from the mushroom Paxillus curtisii. AB - In our continuous investigation for free radical scavengers from extracts of fruit body of basidiomycetes, we have isolated four new p-terphenyl compounds, designated as curtisians A-D, from the methanolic extract of the fruit body of Paxillus curtisii. These compounds were isolated by silica gel and Sephadex LH-20 column chromatographies, preparative-TLC and HPLC, consecutively. The structures of curtisians were assigned as p-terphenyls with substituents of acetyl, benzoyl, phenylbutyryl, 3-hydroxybutyryl and 3-acetoxybutyryl. Curtisians A, B, C and D exhibited inhibitory activity against lipid peroxidation with IC50, values of 0.15, 0.17, 0.24 and 0.14 microg/ml, respectively. PMID- 10805572 TI - Isolation and identification of antifungal N-butylbenzenesulphonamide produced by Pseudomonas sp. AB2. AB - An antifungal bacterial strain, isolated from a greenhouse soil sample, inhibits growth of microflora nearby. It was selected for further studies of bacterial antifungal properties. This isolate was identified as a Pseudomonas sp. based on carbohydrate utilization, and other biochemical and physiological tests. Petri plate assay revealed that the Pseudomonas sp. exhibited antifungal activity against the plant pathogens, Pythium ultimum, Rhizoctonia solani, Phytophthora capsici, Botrytis cinerea and Fusarium oxysporum. Using direct inhibition bioassay on TLC plates after ethyl acetate extraction of the culture filtrate, we correlated antifungal activity with production of antifungal compounds. An antifungal antibiotic was isolated from the culture filtrate and was identified as N-butylbenzenesulphonamide. ED50, values of the N-butylbenzenesulphonamide against P. ultimum, P. capsici, R. solani, and B. cinerea were 73, 41, 33 and 102 ppm, respectively. PMID- 10805571 TI - A new antimitotic substance, FR182877. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activities. AB - Using the characteristic morphological changes of mammalian cells, we screened novel antimitotic substances and found that a strain of Streptomyces sp. No.9885 produced FR182877. This substance was isolated from the culture broth by ethyl acetate extraction, silica gel column chromatography and ODS column chromatography. Structural studies on FR182877 suggested that it had a unique hexacyclic structure encompassing its highly strained double bond. FR182877 exhibited potent antitumor activities against murine ascitic tumor and solid tumor in vivo. PMID- 10805573 TI - Melleolides K, L and M, new melleolides from Armillariella mellea. AB - Three new sesquiterpenoid aromatic esters designated melleolides K (1), L (2) and M (3) were isolated from the cultured mycelia of Armillariella mellea (Vahl. ex Fr.) Karst. Structures of these compounds were determined on the basis of various NMR spectral data, chemical transformations and X-ray analysis. Compounds 1, 2 and 3 showed antimicrobial activities. PMID- 10805574 TI - Chemical characterisation of disruptants of the Streptomyces coelicolor A3(2) actVI genes involved in actinorhodin biosynthesis. AB - The actVI genetic region of Streptomyces coelicolor A3(2) is part of the biosynthetic gene cluster of actinorhodin (ACT), the act cluster, consisting of six ORFs: ORFB, ORFA, ORF1, ORF2, ORF3, ORF4. A newly devised method of ACT detection with a combination of HPLC and LC/MS was applied to the analysis of the disruptants of each ORF. ACT was produced by those of ORFB, ORFA, ORF3, and ORF4. Instead of ACT, the ORF1 disruptant produced 3,8-dihydroxy-1-methylanthraquinone 2-carboxylic acid (DMAC) and aloesaponarin II as shunt products. The ORF2 disruptant gave 4-dihydro-9-hydroxy-1-methyl-10-oxo-3-H-naphtho-[2,3-c]-pyran-3 (S)-acet ic acid, (S)-DNPA. These results support our previous proposal for stereospecific pyran ring formation in the biosynthesis of ACT, most importantly suggesting that the actVI-ORF2 product would recognize (S)-DNPA as a substrate for stereospecific reduction at C-15. The disruptant of ORFA produced (S)-DNPA together with ACT, suggesting that actVI-ORFA might play a role such as stabilising the multicomponent, type II PKS complex. PMID- 10805575 TI - Biosynthesis of cladospirone bisepoxide, a member of the spirobisnaphthalene family. AB - The biosynthesis of cladospirone bisepoxide (1) was investigated by feeding 13C labeled acetate to growing cultures of the fungus Sphaeropsidales sp. (strain F 24'707). 13C NMR spectral analysis demonstrated the polyketide origin of both naphthalene units. The origin of two epoxide oxygens was confirmed as from air by cultivation of the strain in an 18O2-enriched atmosphere. The [18O]incorporation pattern into palmarumycin C12 (11), the putative precursor of 1 led to the hypothesis that the carbonyl oxygen of 1 is derived from water by exchange of an oxygen atom. Inhibition of the biosynthesis of 1 with tricyclazole, an inhibitor of the 1,8-dihydroxynaphthalene (DHN) melanin biosynthesis, confirmed the connection of both biosynthetic pathways. PMID- 10805576 TI - Enhancement of the antifungal activity of rapamycin by the coproduced elaiophylin and nigericin. AB - Streptomyces hygroscopicus ATCC 29253 produces rapamycin, elaiophylin and nigericin. Although elaiophylin has no activity against Candida albicans ATCC 11651, it markedly enhances rapamycin's antifungal activity. Nigericin has only weak activity on its own but it also enhances rapamycin action. Surprisingly, elaiophylin does not enhance nigericin activity on C. albicans. PMID- 10805577 TI - Adxanthromycins A and B, new inhibitors of ICAM-1/LFA-1 mediated cell adhesion molecule from Streptomyces sp. NA-148. II. Physico-chemical properties and structure elucidation. AB - Adxanthromycins A and B are new inhibitors of ICAM-1/LFA-1 mediated cell adhesion molecule isolated from the fermentation broth of Streptomyces sp. NA-148. The molecular formula of adxanthromycins A and B were determined as C42H40O17 and C48H50O22, respectively by FAB-MS and NMR spectral analyses, and the structures of both compounds were elucidated to be a dimeric anthrone peroxide skeleton containing alpha-D-galactose by various NMR spectral analyses and chemical degradation. PMID- 10805578 TI - Synthesis and insecticidal activity of spinosyn analogs functionally altered at the 2'-,3'- and 4'-positions of the rhamnose moiety. AB - In an effort to increase the insecticidal activity of the spinosyn family of naturally occurring macrolides, the 2'-, 3'- and 4'-O-desmethyl-O-acetyl analogs and the 2'-, 3'-, and 4'-O-desmethoxy analogs have been synthesized. These analogs were prepared synthetically from the minor spinosyn factors H, J, K, L and Q either via direct acylation of the corresponding factor or deoxygenation of an intermediate xanthate. The acylated analogs were all more potent insecticides against Heliothis virescens larvae than their respective parent factors, but not as potent as spinosyns A or D. The deoxy analogs were also more potent insecticides than their respective parent factors. The 2'-desmethoxy analogs showed, for the first time, analogs with insecticidal potency against H. virescens greater than that of spinosyns A and D, indicating that polarity is not well tolerated in the rhamnose moiety of spinosyn A. PMID- 10805579 TI - Trichostatin D, a new inducer of phenotypic reversion in transformed cells. PMID- 10805580 TI - Roseoferin, a new aminolipopeptide antibiotic complex from Mycogone rosea DSM 12973, structures and biological activities. PMID- 10805581 TI - Isolation and structural elucidation of AC326-alpha, a new member of the moenomycin group. PMID- 10805582 TI - A trichothecene efflux pump encoded by Tri102 in the biosynthesis gene cluster of Fusarium graminearum. PMID- 10805583 TI - A new plate assay for detecting inhibitors of Candida aspartyl proteinase. PMID- 10805584 TI - A new antimitotic substance, FR182877. II. The mechanism of action. PMID- 10805585 TI - Comparison of the therapeutic efficacy of oral doses of fluconazole and griseofulvin in a guinea pig model of dermatophytosis. PMID- 10805586 TI - CRM646-A and -B, novel fungal metabolites that inhibit heparinase. PMID- 10805587 TI - Gibberellin biosynthesis: its regulation by endogenous and environmental signals. AB - The hormone gibberellin (GA) plays an essential role in many aspects of plant growth and development, such as seed germination, stem elongation and flower development. In recent years, exciting progress has been made in understanding how the biosynthesis of this hormone is regulated by endogenous and environmental factors. This has resulted from isolation of genes encoding enzymes involved in GA biosynthesis and metabolism, which also enabled us to manipulate the pathway by modifying the expression of these genes in transgenic plants. In addition, new GA response mutants provided information about how signaling components are involved in feedback regulation of the GA biosynthetic pathway. PMID- 10805589 TI - Purification from conditioned medium and chemical identification of a factor that inhibits somatic embryogenesis in carrot. AB - Somatic embryogenesis is strongly inhibited in cultures of carrot (Daucus carota L.) cells when the cell density is high. The inhibition is caused by factors that are released by cells into the medium of such cultures. In this study, we purified and identified one of the inhibitory factors found in the medium of high cell-density cultures of carrot cells. The inhibitory factor with the strongest apparent activity was purified by fractionation with ethylacetate, chromatography on an octadecylsilyl (ODS) silica gel-column and HPLC. The inhibitory factor had a single peak of absorbance at 280 nm and was identified as 4-hydroxybenzyl alcohol by mass spectrometry and 1H- and 13C-NMR spectroscopy. Authentic 4 hydroxybenzyl alcohol strongly inhibited the formation of somatic embryos at a concentration equal to that in high-cell-density cultures. These results suggest that 4-hydroxybenzyl alcohol is a major factor that accumulates in high-cell density cultures of carrot cells and inhibits somatic embryogenesis. PMID- 10805588 TI - Novel mannose-binding rice lectin composed of some isolectins and its relation to a stress-inducible salT gene. AB - The novel mannose-binding rice lectin (MRL) purified by Sephadex G-50 or maltamyl Sepharose 4B affinity chromatography was not homogeneous, but the components were separated clearly by two dimensional polyacrylamide gel electrophoresis (1st; isoelectric focusing with Immobiline, 2nd; SDS-PAGE). The major spots were located at pI 4.85 and 4.74, and minor spots at pI 4.66, 4.56, and 4.44; all spots were distributed at about MW 45,000. Other faint spots were sometimes detected just below the major spots. In the western blot analysis, all the spots reacted with the monoclonal antibodies specific to MRL, which bound to MRL and inhibited the lectin activity to agglutinate rabbit erythrocytes. The proteins of the spots at pI 4.85, 4.77, 4.66, and 4.56 had lectin activity. The major proteins at pI 4.85 and 4.77 also had the common amino acid sequence at N terminus, TLVKIGPWGGNGGSAQDISV, which is almost identical to salt and drought stress-inducible salT gene products in rice plants. High homology was also conserved in both the cDNA and the genomic clones encoding the MRL component at pI 4.85, which were selected with MRL-specific antibodies and an oligonucleotide designed from the partial amino acid sequence. All results suggest that MRL is composed of several isolectins, if not, related proteins having a common epitope and may belong to a family of stress-inducible proteins. PMID- 10805590 TI - Two novel transcripts expressed in pea dormant axillary buds. AB - To elucidate the molecular mechanism of apical dominance, the expression patterns of genes that are preferentially expressed in dormant axillary buds of pea (Pisum sativum L. cv. Alaska) seedlings were investigated. We isolated two cDNA clones, cPsAD1 and cPsAD2 whose corresponding genes were named PsAD1 and PsAD2, from a cDNA library of dormant axillary buds using the differential display method. The deduced amino acid sequence of PsAD1 contains 87 residues and is rich in glycine residues in the amino terminal region. A search of the protein databases failed to find any sequences similar to PsAD1 protein except for the glycine-rich region. Northern blot analyses showed that PsAD1 mRNA mainly accumulated in dormant axillary buds and that its amount rapidly decreased after decapitation of the terminal bud. In situ hybridization analyses indicated that PsAD1 mRNA was localized in the apical meristem, procambia, and leaf primordia in dormant axillary buds that were competent to grow out but whose growth was temporarily suspended. That is, the expression of the PsAD1 gene is closely associated with the dormancy of axillary buds. The deduced amino acid sequence of PsAD2 contains 98 amino acid residues and is not similar to those of previously characterized proteins. PsAD2 mRNA accumulated in dormant axillary buds, roots, mature leaflets and elongated stems, suggesting that PsAD2 is involved in not only the dormancy of axillary buds but also the non-growing state in various tissues. PMID- 10805591 TI - An RNA-binding protein, AtRBP1, is expressed in actively proliferative regions in Arabidopsis thaliana. AB - A cDNA clone, named XF41, that encodes an RNA-binding protein was isolated from Arabidopsis thaliana. The deduced protein, named AtRBP1, contains two conserved consensus sequence-type RNA-binding domains (CS-RBDs) in the N-terminal half, a putative PY motif (a target of a WW domain) in the center, and uncharacterized C terminal domain. A binding assay demonstrated that the AtRBP1 can bind to single stranded nucleic acids in vitro. Analysis of localization of the GUS activity of transgenic Arabidopsis thaliana plants that have the chimeric gene containing the upstream sequence of the AtRBP1 gene and GUS gene demonstrated that the AtRBP1 gene is expressed in meristematic tissues such as the vegetative shoot apex and root tips, developing organs such as floral buds and pistils of young flowers, abscission layers of immature siliques and junctions of pedicels. Considering the specificity of the expression, AtRBP1 may be required in the progress of cell proliferation. PMID- 10805592 TI - Stoichiometric analysis of barley plastid ribosomal proteins. AB - We analyzed the protein composition of plastid 70S ribosomes isolated from the stromal fractions of barley plastids by the radical-free and highly reducing method of two dimensional polyacrylamide gel electrophoresis (RFHR 2D-PAGE). Intactness of the ribosomes was confirmed by the poly(U)-directed phenylalanine polymerization activity and by the reassociation capacity of the subunits into 70S ribosomes. The small and large ribosomal subunits were composed of 23 and 36 proteins, respectively. In addition, one acidic protein associated with ribosomes in low salt buffer but released in high salt buffer was found. The plastid ribosomes contained relatively larger numbers of acidic proteins than prokaryotic ribosomes. Stoichiometric analysis revealed the presence of several ribosomal proteins in low copy numbers, indicating that the ribosomes of plastids were heterogeneous. We also investigated the protein composition of plastid ribosomes from greening barley leaves and found that it did not change during greening. PMID- 10805593 TI - Inhibition of cell-plate formation by brefeldin A inhibited the depolymerization of microtubules in the central region of the phragmoplast. AB - Treatment of tobacco BY-2 cells with 20 microM brefeldin A (BFA), which causes disassembly of the Golgi apparatus (Yasuhara et al. 1995), completely inhibited the formation of the cell plate when the treatment was started before the chromosomes had begun to to condense. In cells in which cell-plate formation was inhibited by BFA, the centrifugal development of the phragmoplast was also inhibited. In such cells, the depolymerization of microtubules in the central region of the phragmoplast did not occur at least for 1 h after the formation of the phragmoplast, while the centrifugal development of the phragmoplast and cell plate formation were completed in almost all cells not treated with BFA. The inhibition of cell-plate formation seems to inhibit the centrifugal development of the phragmoplast by inhibiting the depolymerization of microtubules in the central region of the phragmoplast, which is required for the supply of free tubulin necessary for the polymerization of microtubules at the outer margins of the phragmoplast. PMID- 10805594 TI - Evaluation of the defense system in chloroplasts to photooxidative stress caused by paraquat using transgenic tobacco plants expressing catalase from Escherichia coli. AB - We evaluated the defense system in chloroplasts to photooxidative stress imposed by paraquat treatment under illumination in transgenic tobacco plants with increased tolerance to drought stress at a high light intensity produced by catalase from Escherichia coli targeted to chloroplasts [Shikanai et al. (1998) FEBS Lett. 428: 47]. At 24 h after the paraquat application, Chl was destroyed in the wild-type plants, but not in transgenic plants. Photosynthetic activities monitored by CO2 fixation and Chl fluorescence were much less affected by the paraquat treatment in transgenic lines. The activities of chloroplastic ascorbate peroxidase (APX) isoenzymes decreased in parallel with the depletion of ascorbate (AsA) in leaves in both lines. Paraquat treatment had no effect on the transcript level of chloroplastic APX isoenzymes, while it significantly lowered the level of their proteins. These data suggest that the depletion of AsA in chloroplasts under severe stress conditions inactivates and degrades chloroplastic APX isoenzymes. PMID- 10805595 TI - Post-transcriptional gene silencing in cultured rice cells. AB - Post-transcriptional gene silencing (PTGS) has been shown to occur in many transgenic dicotyledonous plants. Although transgene silencing has been documented in transgenic monocots, PTGS has been only recently described in monocotyledonous plants. To study PTGS in rice, we generated 25 transgenic rice cell lines in which the 35S-gus gene was stably integrated. In one transgenic cell line, lack of gus expression was shown to be caused by PTGS based on the results of run-on transcription assays. Furthermore, to examine whether reintroduction of the same plasmid DNA into cells in which the gus gene was highly expressed causes the suppression of the gus expression, the gus plasmid was introduced into protoplasts isolated from a high gus-expressing line. Results of the kinetic analysis of the GUS activities in transfected protoplasts indicated that the gus expression in transfected protoplasts was suppressed by the gus plasmid but not by the luc plasmid harboring the 35S-luciferase gene, suggesting that suppression of the gus expression by introduced plasmid DNA was homology-dependent. These results suggest that the transgenic protoplasts may be a useful experimental system to study PTGS in rice and other species. PMID- 10805596 TI - Ethylene formation and phenotypic analysis of transgenic tobacco plants expressing a bacterial ethylene-forming enzyme. AB - A bacterial ethylene-forming enzyme (EFE) catalyzes oxygenation of 2-oxoglutarate to produce ethylene and carbon dioxide in contrast to a plant enzyme which uses 1 aminocyclopropane-1-carboxylic acid as a substrate. We constructed several lines of transgenic tobacco plants which expressed an EFE from Pseudomonas syringae pv. phaseolicola PK2. The gene encoding a chimeric protein consisting of EFE and beta glucuronidase (GUS) was introduced into the tobacco genome using a binary vector which directs expression of the EFE-GUS fusion protein under the control of constitutive promoter of cauliflower mosaic virus 35S RNA. Two lines of transgenic plants produced ethylene at consistently higher rates than the untransformed plant, and their GUS activities were expressed in different tissues. A significant dwarf morphology observed in the transgenic tobacco displaying the highest ethylene production resembled the phenotype of a wild-type plant exposed to excess ethylene. These results demonstrate a potential use of bacterial EFE to supply ethylene as a hormonal signal via an alternative route using an ubiquitous substrate 2-oxoglutarate in plant tissues. PMID- 10805597 TI - Determination of the rate of photoreduction of O2 in the water-water cycle in watermelon leaves and enhancement of the rate by limitation of photosynthesis. AB - A study was performed to determine how the electron fluxes for the photosynthetic carbon reduction (PCR) and the photorespiratory carbon oxidation (PCO) cycles affect the photoreduction of O2 at PSI, which is the limiting step in the water water cycle. Simultaneous measurements were made of CO2-gas exchange, transpiration and quantum yield of PSII [phi(PSII)] using leaves of watermelon (Citrullus lanatus). The total electron flux in PSII[Je(PSII)], as estimated from phi(PSII), was always larger than the total electron flux required for the PCR and PCO cycles at various partial pressures of CO2 and O2 and 1,100 micromol photons m(-2)s(-1). This observation suggested the existence of an alternative electron flux (Ja). Ja was divided into O2-dependent [Ja(O2-depend)] and O2 independent [Ja(O2-independ)] components. The magnitude of half Ja(O2-depend), 7.5 to 9.5 micromol e- m(-2)s(-1), and its apparent Km for O2, about 8.0 kPa, could be accounted for by the photoreduction of O2 at PSI either mediated by ferredoxin or catalyzed by monodehydroascorbate reductase. The results indicated that Ja(O2-depend) was driven by the water-water cycle. A decrease in the intercellular partial pressure of CO2 from 23 to 5.0 Pa at 21 kPa O2 enhanced Ja(O2-depend) by a factor of 1.3. Saturation of the activities of both the PCR and PCO cycles by increasing the photon flux density induced Ja. These results indicate the electron flux in PSII that exceeds the flux required for the PCR and PCO cycles induces the photoreduction of O2 in the water-water cycle. PMID- 10805598 TI - Photosynthesis during desiccation in an intertidal alga and a land plant. AB - This study was undertaken to determine how photosynthesis tolerates desiccation in an intertidal alga Fucus vesiculosus L. and a terrestrial sunflower Helianthus annuus L. Photosynthetic O2 evolution generally was inhibited at low water potentials (psiw) but more in sunflower leaves than in Fucus fronds at the same psiw. As psiw decreased, less carbon accumulated in an organic carbon store in Fucus. The inhibition of photosynthesis appeared to be mostly biochemical because it could not be prevented by supplying additional CO2 or by supplying CO2 from the internal organic carbon store. The inhibition of photosynthesis and carbon storage occurred after turgor disappeared and thus when solute concentrations were increasing in the cells. Solute concentrations were much higher in Fucus than in sunflower. After desiccation to the air-dry state (psiw below - 10 MPa), photosynthesis could not recover in sunflower but it recovered rapidly when Fucus was exposed to seawater. The lack of recovery in sunflower was associated with inability to recover turgor probably because of breaks in cell membranes. The ability to recover in Fucus was gradually lost during 1.5 d of desiccation at 45% relative humidity. At lower humidities, recovery was lost sooner as small amounts of water were removed. We conclude that photosynthesis tolerated desiccation more in Fucus than in sunflower because of differences in the molecular environment around the photosynthetic enzymes. Important aspects of this environment were features that prevented membrane breakage but promoted the retention of small amounts of water that were critical for viability. PMID- 10805599 TI - Hormonal cross-talk between auxin and ethylene differentially regulates the expression of two members of the 1-aminocyclopropane-1-carboxylate oxidase gene family in rice (Oryza sativa L.). AB - Two cDNA clones, pOS-ACO2 and pOS-ACO3, encoding 1-aminocyclopropane-1 carboxylate (ACC) oxidase were isolated from rice seedling cDNA library. pOS-ACO3 is a 1,299 bp full-length clone encoding 321 amino acids (Mr=35.9 kDa), while pOS ACO2 is 1,072 bp long and is a partial cDNA clone encoding 314 amino acids. These two deduced amino acid sequences share 70% identity, and display a high degree of sequence identity (72-92%) with previously isolated pOS-ACO1 of deepwater rice. The chromosomal location studies show that OS-ACO2 is positioned on the long arm of chromosome 9, while OS-ACO3 on the long arm of chromosome 2 of rice genome. A marked increase in the level of OS-ACO2 transcript was observed in IAA-treated etiolated rice seedlings, whereas the OS-ACO3 mRNA was greatly accumulated by ethylene treatment. Results of ethylene inhibitor studies indicated that auxin promotion of the OS-ACO2 transcription was not mediated through the action of auxin-induced ethylene. Thus, it appears that there are two groups of ACC oxidase transcripts in rice plants, either auxin-induced or ethylene-induced. The auxin induced OS-ACO2 expression was partially inhibited by ethylene, while ethylene induction of OS-ACO3 transcription was completely blocked by auxin. These results indicate that the expression of ACC oxidase genes is regulated by complex hormonal networks in a gene specific manner in rice seedlings. Okadaic acid, a potent inhibitor of protein phosphatase, effectively suppressed the IAA induction of OS-ACO2 expression, suggesting that protein dephosphorylation plays a role in the induction of ACC oxidase by auxin. A scheme of the multiple regulatory pathways for the expression of ACC oxidase gene family by auxin, ethylene and protein phosphatase is presented. PMID- 10805600 TI - Boron nutrition of cultured tobacco BY-2 cells. III. Characterization of the boron-rhamnogalacturonan II complex in cells acclimated to low levels of boron. AB - Cultured cells of tobacco (Nicotiana tabacum L.) BY-2 which could propagate at the same rate as the parent cells (1 mg B liter(-1)) under a lower level of boron (0.01 mg B liter(-1)) were obtained. The selected cells had swollen cell walls. In the parent cells, all the RG-II occurred as a B-RG-II complex, however, two thirds of the RG-II occurred in a monomeric form in the selected cells. PMID- 10805601 TI - Chloroplast tubules visualized in transplastomic plants expressing green fluorescent protein. AB - A fusion between the plastid psbA promoter and the green fluorescent protein gene (gfp) was introduced into the tobacco chloroplast genome by stable plastid transformation. GFP was synthesized actively and exclusively in the chloroplasts. Tubular projections filled with GFP but containing no chlorophyll were visualized for the first time in chloroplasts of these transplastomic plants. Occasionally, the tubules connect chloroplasts with each other, suggesting the possibility of the exchange of endogenous proteins. However, the fusion of protoplasts between the transplastomic and wild-type plants showed that such chloroplast connections might be rare in mesophyll protoplasts. PMID- 10805602 TI - Does aluminum inhibit pollen germination via extracellular calmodulin? AB - The effect of aluminum (Al) on pollen germination and its mechanism of action were investigated. Pollen germination and pollen tube elongation were inhibited by Al at pH 4.5. This inhibitory effect was reversed by the addition of purified calmodulin (CaM), whereas neither the calcium binding-protein S-100 nor Al chelator citric acid at the same concentrations had any obvious effect on Al inhibited pollen germination. The presence of either the membrane-impermeable CaM inhibitor anti-CaM antiserum or Ca2+ chelator EGTA completely suppressed the effect of exogenous CaM. These results indicate the involvement of extracellular calmodulin in the short-term effects of Al on pollen germination and pollen tube elongation. PMID- 10805603 TI - Zinc-finger genes that specifically express in pistil secretory tissues of petunia. AB - Tissue-specific expression patterns of petunia zinc-finger genes, ZPT2-10 and ZPT3-3, were analyzed by using GUS reporter system. The GUS expression directed by ZPT2-10 promoter was specifically found in the stylar transmitting tissue of pistil, and that by ZPT3-3 promoter in stigmatic and stylar transmitting tissues. These tissues play important roles in reproductive process. We discuss possible roles of the zinc-finger proteins in these specialized tissues. PMID- 10805604 TI - Response of nicotine self-administration in the rat to manipulations of mu-opioid and gamma-aminobutyric acid receptors in the ventral tegmental area. AB - RATIONALE: The mesolimbic dopamine system has been implicated in the reinforcing effects of nicotine, a drug which appears to act at least in part through the ventral tegmental area (VTA). Other neuronal elements in the VTA are important in drug reward. In particular, mu opioid receptors in the VTA have been shown to influence cocaine reinforcement. OBJECTIVE: The aim of this study was to test whether the mu opioid receptors in the VTA also regulate the intake of nicotine. METHODS: This research was carried out with animals trained to self-administer nicotine or cocaine, or to respond for food. Mu receptors were targeted with the selective agonist [D-Ala2,N-Me-Phe4,Gly-ol5]-enkephalin (DAMGO) and gamma aminobutyric acid (GABA) receptors with the selective agonists baclofen and muscimol; each of these compounds was delivered by microinfusion into the VTA. RESULTS: The mu-selective agonist DAMGO, tested over a dose range of 0.005-0.05 microg, had an effect at the highest dose only, where it produced a reduction in self-administration maintained by doses of either 10 microg/kg or 30 microg/kg per infusion of nicotine. Intra-VTA microinfusions of DAMGO did not reinstate extinguished responding previously established for nicotine, nor did they have prominent effects on operant behavior maintained by food. In contrast to the overall limited effects of DAMGO on nicotine self-administration, the GABA agonists muscimol and baclofen each reduced nicotine self-administration substantially when delivered into the VTA, whereas they were less effective against cocaine self-administration. CONCLUSIONS: The lesser effect of DAMGO microinfusions in the VTA on nicotine than cocaine self-administration is associated with the opposite efficacy of GABA agonists. These findings suggest that nicotine and cocaine differentially activate circuitry in which mu receptors are situated, especially GABAergic elements. PMID- 10805605 TI - Opiate withdrawal-induced place aversion lasts for up to 16 weeks. AB - RATIONALE: Administration of low doses of opiate antagonists to morphine dependent rats produces an aversive response as measured by a conditioned place aversion, but the time course of such a learned aversion is largely unknown. OBJECTIVES: The purpose of this experiment was to examine the time course for the expression of a place aversion to opiate withdrawal. METHODS: Morphine-dependent rats were tested in a three-chamber place-aversion apparatus. The conditioning phase consisted of three pairings of either naloxone (15 microg/kg s.c.) or vehicle with two compartments, with the most similar time allotments during the preconditioning test. During the testing phase, rats were again allowed to explore the entire apparatus. Different groups were tested at 24 h, 1 week, 2 weeks, 4 weeks, 8 weeks, and 16 weeks post-conditioning (morphine-free tests). RESULTS: A robust place aversion was recorded at every time point tested, including at 16 weeks. In previously published work, placebo-pelleted rats tested with naloxone at the same dose failed to show a place aversion and nondependent rats showed a stable lack of aversion at tests up to 56 days. Dependent animals without naloxone also failed to show a place aversion at any of those time points. CONCLUSIONS: In the absence of any active intervention, the place aversion produced by opiate withdrawal is very long lasting and provides a model for protracted abstinence that may be useful for delineating the neurobiological substrate for vulnerability to relapse. PMID- 10805606 TI - Precipitated and spontaneous withdrawal in rats tolerant to anandamide. AB - RATIONALE: There is evidence that cannabinoids cause tolerance and physical dependence in humans and animals. OBJECTIVES: The aim of this work was to study whether the endogenous ligand for the cannabinoid receptor, arachidonylethanolamide (anandamide), induced behavioral tolerance and physical dependence in rats. METHODS: Rats were injected with anandamide (20 mg/kg i.p.) daily for 2 weeks. To assess tolerance, on days 1, 8 and 15 of treatment rats were observed and behavior was tested. Two common methods were employed to assess physical dependence: interruption of anandamide dosing and vehicle substitution or administration of the cannabinoid CB1 receptor antagonist SR141716A (3 mg/kg i.p.). RESULTS: Full or partial tolerance developed to the classical behavioral effects elicited by the cannabinoids: hypothermia, catalepsy, hypomotility, decrease in stereotypic activity (rearing and grooming) and hindlimb splaying. No tolerance to anandamide was observed for reduced defecation. An abstinence syndrome appeared after abrupt cessation of cannabinoid intake and after withdrawal precipitated by SR141716A; the withdrawal signs were scratching, licking and biting, eating of feces, ptosis, arched back, wet dog shakes, head shakes, myoclonic spasms, writhing, forepaw fluttering, teeth chattering and piloerection. CONCLUSIONS: These findings indicate that the endogenous cannabinoid ligand, administered exogenously, induces both tolerance and physical dependence in rats. PMID- 10805607 TI - The effects of 3-week estrogen hormone replacement on cognition in elderly healthy females. AB - RATIONALE: Estrogen concentrations decline with age and menopause is often followed by an acceleration of the age effects on cognition. It is suggested that replacement of estrogen would reinstate, at least in part, cognitive abilities. Effects of estrogens on memory have been reported in studies with women in a clinical setting who either needed or wished to have the estrogen replacement and are mostly in the perimenopausal age-band. OBJECTIVE: The present study investigated the effects of estradiol on memory and on frontal lobe function in elderly female subjects who did not suffer any of the postmenopausal symptoms and had never taken estrogen hormone replacement (EHR) previously. METHODS: EHR (Progynova TS, transdermal estradiol; n=19) or placebo (n=18) was given for a period of 3 weeks to elderly healthy female subjects. Memory, frontal lobe functions (inhibition and planning) and visuospatial abilities (mental rotation) were tested before and after treatment. Estrogen plasma levels were measured to confirm the result of EHR. Cortisol plasma levels were also measured before and after cognitive performance in order to evaluate the effects of EHR on the sensitivity of the hypothalamo-pituitary-adrenal (HPA) axis to mild mental stress. RESULTS: Plasma estradiol levels in the drug group increased to levels equivalent to that of a fertile woman (0.21+/-0.5 nmol/l). Memory function as well as visuospatial abilities as measured by a mental rotation task improved significantly with EHR. However, there was no effect of EHR on frontal lobe functions. The cognitive effects were not dependent on an improvement in mood or general well-being as may be the case with EHR in women at peri- or post menopausal stage. EHR was found to increase the HPA response to task-induced stress, as indicated by an increase in cortisol plasma levels. CONCLUSIONS: The present study has provided evidence of a beneficial effect of EHR on cognitive abilities given for first time to healthy elderly women. Furthermore, the present study has demonstrated a differential effect of EHR on memory, visuospatial abilities and frontal lobe function. PMID- 10805608 TI - Antagonism of stimulus properties of nicotine by dihydro-beta-erythroidine (DHbetaE) in rats. AB - RATIONALE: Previous work has shown that a dose of DHbetaE, a competitive nicotinic receptor antagonist that blocked the discriminative stimulus properties of nicotine, was insufficient to block locomotor depression or operant rate reducing effects of nicotine in rats. Examination of DHbetaE against other behavioural effects of nicotine may help in understanding its diverse actions. OBJECTIVE: The present experiments examine the aversive stimulus properties of nicotine, a function implicated in the regulation of nicotine intake. Furthermore, to characterise the duration of pharmacological blockade produced by DHbetaE, the antagonist was examined in the drug discrimination (DD) procedure. METHODS: Using the conditioned taste aversion (CTA) paradigm, male hooded rats were trained to avoid one of two distinctively flavoured solutions paired with nicotine (0.2 or 0.4 mg/kg) administration. In rats trained to discriminate 0.2 mg/kg s.c. nicotine in a two-lever procedure maintained under a tandem V160'' FR10 schedule of food reinforcement, the offset of antagonism by DHbetaE was examined 5, 15 and 30 min following injection of nicotine (0.2 or 0.4 mg/kg s.c.) or vehicle. RESULTS: Administration of DHbetaE (0.5, 1.6 and 5.0 mg/kg s.c.) 30 min before nicotine failed to block nicotine (0.4 mg/kg) CTA, while co administration of DHbetaE (5.0 mg/kg s.c.) with nicotine (0.2 and 0.4 mg/kg s.c.) prevented the development of CTAs. This blockade complemented nicotine discrimination data in which DHbetaE blocked the discriminative stimulus effect of nicotine (0.2 or 0.4 mg/kg s.c.) for 45 min after its administration. CONCLUSIONS: These observations of DHbetaE's short-lasting antagonism against the aversive and discriminative stimulus effects of nicotine support the involvement of the similar subtypes of nicotinic receptor in the mediation of these diverse behavioural effects. PMID- 10805609 TI - Impulsiveness and the prolactin response to d-fenfluramine. AB - RATIONALE: A number of studies have reported abnormalities of serotonin function in aggressive and impulsive behaviours in psychiatric and forensic populations. It is unknown whether this is because serotonin function plays a part in determining the dimension of trait impulsiveness in the general population or whether this is restricted to these behaviourally extreme groups. METHOD: The prolactin response to d-fenfluramine was measured in subjects scoring high and low on a scale of impulsiveness selected from a panel of healthy volunteers screened for impulsiveness. Measures included the 17 impulsiveness scale, State Trait Anger Expression Inventory (STAngXI) and the Tridimensional Personality Questionnaire (TPQ). Plasma cortisol was also determined along with fenfluramine and its metabolite norfenfluramine. RESULTS: The high impulsive group had reduced AUC (PRL) compared with the low impulsive group; this remained significant after adjusting for baseline prolactin, cortisol and drug levels. There was no significant association between impulsiveness, the harm avoidance subscale of the TPQ or trait anger (STAngXI) and prolactin rise. Repeated serum prolactin measures were not significantly different between the two groups. CONCLUSIONS: This study provides some support to the hypothesis that reduced serotonin function contributes to high trait impulsiveness and is not restricted to behaviourally extreme populations. PMID- 10805610 TI - Comparison of intravenous cocaethylene and cocaine in humans. AB - RATIONALE: Cocaethylene is a pharmacologically active homolog and metabolite of cocaine, formed by transesterification of cocaine in the presence of ethanol. Here we relate findings from a randomized, placebo-controlled, double-blind study in which we examined the physiological and subjective effects and pharmacokinetics of i.v. administered cocaethylene in human volunteers using cocaine as a comparator. METHODS: Cocaine-dependent participants randomly received one study drug, cocaethylene (0.25 or 0.5 mg/kg), cocaine (0.25 or 0.5 mg/kg), or placebo, during each experimental session which occurred on separate days. RESULTS: Cocaethylene was less potent in elevating heart rate than equivalent doses of cocaine. Similar differences between cocaine and cocaethylene were found for subjective measures ("Cocaine High", "Rush", "Stimulated" and "Good Drug Effects"). All active drug conditions produced significant increases in systolic blood pressure relative to placebo, but no significant effect on diastolic blood pressure was observed. Cocaethylene demonstrated a slower clearance, larger volume of distribution and correspondingly longer elimination half-life than cocaine. CONCLUSION: The findings from this study confirm those of previous studies that show that cocaethylene has pharmacological properties in common with cocaine, but is less potent. PMID- 10805611 TI - Effect of coadministration of clozapine and fluvoxamine versus clozapine monotherapy on blood cell counts, plasma levels of cytokines and body weight. AB - RATIONALE: Clozapine treatment is associated with side-effects such as blood cell dyscrasias and weight gain. Increased plasma levels of the cytokines and soluble cytokine receptors leptin, tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptors p55 and p75, as well as toxic metabolites of clozapine, have been suggested as the basis for these side-effects, OBJECTIVES: This study examined whether the coadministration of the selective serotonine reuptake inhibitor fluvoxamine, which interferes with clozapine's hepatic metabolism, affects the immunomodulation by clozapine and some of its side-effects. METHODS: The following parameters were measured: circulating levels of the cytokines and soluble receptors, plasma concentrations of clozapine and its metabolite N desmethylclozapine, body weight and blood cell counts in 11 and 12 schizophrenic inpatients on combined and monotherapy, respectively, before and during the first 6 weeks of medication. RESULTS: On the basis of comparable plasma levels of clozapine and N-desmethylclozapine, the coadministration of fluvoxamine 1) attenuated and delayed the clozapine-induced increase in TNF-alpha plasma levels, 2) enhanced and accelerated the clozapine-induced increase in leptin plasma levels without significant effect on clozapine-induced weight gain, and 3) decreased granulocyte counts. CONCLUSIONS: As clozapine, its metabolite N desmethylclozapine and fluvoxamine are unlikely to make these differences, other metabolites might be responsible. The coadministration of clozapine and fluvoxamine offers the opportunity to investigate further the putative associations between certain metabolites, immunomodulation and these side effects. PMID- 10805612 TI - Sex differences in the discriminative stimulus effects of m chlorophenylpiperazine and ethanol withdrawal. AB - RATIONALE: The serotonergic system plays a role in regulation of anxiety and ethanol withdrawal (EW). Nevertheless, few studies have assessed sex differences in serotonergic effects on EW. OBJECTIVES: This study examined sex differences in the anxiogenic stimuli induced by a serotonin (5-HT)(1b,2) agonist, meta chlorophenylpiperazine (mCPP), prior to ethanol and during EW. METHODS: Gonadectomized or sham-operated adult male and female rats and 17beta-estradiol (2.5 mg, 21-day release, s.c.) -replaced ovariectomized (OVX) rats were trained to discriminate mCPP (1.2 mg/kg, i.p.) from saline in a two-lever choice task for food. Latency to the first lever press and mCPP lever selection were measured following mCPP (0-1.2 mg/kg). Rats then received chronic ethanol-containing liquid diet (6.5%) for 10 days and were tested for mCPP lever selection 12 h and 36 h after removal of ethanol. RESULTS: Fewer sham female and beta-estradiol replaced OVX rats selected the mCPP lever than male or OVX rats, and showed an increased initiation latency after mCPP injection. During EW (12 h and 36 h), fewer sham female and beta-estradiol-replaced OVX rats responded on the mCPP lever after saline injection as well as after mCPP challenge than male or OVX rats. Castration did not alter any response of male rats to mCPP. CONCLUSIONS: (1) mCPP discrimination is a useful measure of EW in male and female rats; and (2) sham female and beta-estradiol-replaced OVX rats are less sensitive to the discriminative stimulus prior to and during EW, but more sensitive to impaired behavioral initiation induced by mCPP than male or OVX rats. PMID- 10805613 TI - Comparison of benzodiazepines and the non-benzodiazepine agents zolpidem and zaleplon with respect to anxiolytic action as measured by increases in hypertonic NaCl-solution drinking in rats. AB - RATIONALE: In previous studies, water-deprived rats offered hypertonic 1.5% NaCl solutions to drink showed increased intakes when treated with agents known to have anxiolytic action in humans. This study explored two non-benzodiazepine (non BZ) sedative-hypnotic agents, zolpidem and zaleplon, and compared them with three traditional BZs. OBJECTIVES: Although many studies confirm that treatment with BZs possessing sedative-hypnotic and anxiolytic actions also produces acute increases in food and fluid ingestion in animals, zolpidem has yielded conflicting results. To help resolve this question, we compared three BZs with zolpidem and zaleplon with respect to their actions in increasing the ingestion of 1.5% NaCl solution in water-deprived rats. METHODS: Rats were adapted to a water-deprivation schedule permitting drinking for 1 h daily. Once or twice each week, 1.5% NaCl solution was substituted for water during the drinking session and, 15 min pre-session, rats were given a drug or vehicle dose by gavage (p.o.) to delineate the dose-effect relationships for zolpidem, zaleplon, alprazolam, clonazepam, and chlordiazepoxide. Then, the dose-effect relationship for zolpidem was re-determined. A second study with two groups, using both zolpidem and clonazepam, explored whether following the dose-effect determination of a drug by a second determination affected the second relationship, and whether dose-effect determinations of either agent affected the results of the second agent investigated. RESULTS: All agents yielded dose-related increases in 1.5% NaCl solution ingestion, except the first zolpidem determination in the first study. In the second study, all determinations yielded dose-related increases, with no indication that the set of determinations for the first agent affected those for the second agent. CONCLUSIONS: The BZ and non-BZ agents explored yielded significant dose-effect relationships using this procedure, confirming their classification among the anxiolytic agents. The initial negative result for zolpidem in the first study may indicate a less reliable anxiolytic action for this agent, although this could not be resolved as attributable to drug history. PMID- 10805615 TI - Clozapine discrimination with a low training dose distinguishes atypical from typical antipsychotic drugs in rats. AB - RATIONALE: Previous drug discrimination studies with clozapine have not reliably distinguished between atypical and typical antipsychotics. OBJECTIVES: The present study was conducted to determine whether low-dose clozapine drug discrimination could distinguish atypical from typical antipsychotics. METHODS: Rats were trained to discriminate 1.25 mg/kg clozapine from vehicle in a two lever drug discrimination procedure. RESULTS: Generalization testing revealed full substitution with the atypical antipsychotics olanzapine (90.3% maximum generalization), sertindole (99.8%), and risperidone (87.1%) and partial substitution for quetiapine (seroquel, 66.4%) and the typical antipsychotics haloperidol (56.8%) and thioridazine (74.3%). Remoxipride (23.1%) and the typical antipsychotics chlorpromazine (27.9%) and fluphenazine (29.5%) did not reliably substitute for clozapine. CONCLUSIONS: In contrast to previous clozapine drug discrimination studies with higher training doses, the atypical antipsychotics olanzapine, sertindole, and risperidone reliably substituted for clozapine while typical antipsychotics did not. These results suggest that low-dose clozapine drug discrimination may be a more sensitive assay for distinguishing atypical from typical antipsychotic drugs. PMID- 10805614 TI - Role of dopamine in prepulse inhibition of acoustic startle. AB - RATIONALE: Prepulse inhibition of acoustic startle is the reduction in startle response to an intense auditory stimulus when this stimulus is immediately preceded by a weaker prestimulus. Prepulse inhibition occurs normally in humans and experimental animals, but schizophrenic persons often exhibit a marked impairment in this measure. Previous studies have shown that dopamine (DA) dependent neuronal mechanisms are involved in the modulation of prepulse inhibition. OBJECTIVE: Experiments were conducted in rats to elucidate further the involvement of DA-ergic mechanisms in prepulse inhibition. RESULTS: In line with previous studies, the indirect DA agonist, amphetamine, was shown to decrease prepulse inhibition. A close reverse relationship over time between DA overflow in the nucleus accumbens and prepulse inhibition was obtained using a technique allowing concomitant measurement of these parameters in awake, freely moving rats. This effect was more pronounced in amphetamine-treated rats compared to rats treated with equimolar doses of cocaine, which increased DA overflow without affecting prepulse inhibition. In other experiments, the combined treatment with subthreshold doses of the selective DA D1 agonist, SKF 38393, and the selective DA D2 agonist, quinpirole, was also shown to decrease prepulse inhibition. Finally, the selective DA D2 antagonist, raclopride, was shown to enhance prepulse inhibition. CONCLUSIONS: In line with previous studies, it is concluded that DA neurotransmission is involved in the modulation of prepulse inhibition and that the ventral part of the mesostriatal DA system may serve an important role in this modulation. Furthermore, the possibility is discussed that the discrepant results on prepulse inhibition obtained with amphetamine and cocaine may disclose functionally relevant differences in their mechanisms of action, and that the enhancement of prepulse inhibition induced by some antipsychotics in rats may reflect their propensity to induce adverse mental effects in humans. PMID- 10805616 TI - Double-blind placebo-controlled pilot study of paroxetine for specific phobia. AB - Drugs are not recognized as a standard treatment for specific phobia, despite its apparent similarities to other kinds of phobia. Reluctance on the part of patients and clinicians to see the disorder as more than normal anxiety may explain the apparent resistance to pharmacotherapy. Eleven patients fulfilling DSM-IV criteria for specific phobia were randomized to 4 weeks of double-blind treatment with placebo or paroxetine up to 20 mg/day. They were assessed weekly with the Fear Questionnaire and the Hamilton Rating Scale for Anxiety. Paroxetine showed significant superiority in reducing all measures (ANCOVA for reductions in phobia scores F=7.9, P=0.02). One out of six patients responded to placebo, compared to three out of five patients on paroxetine. This new therapeutic option (i.e. drug treatment) for specific phobia deserves further examination in a larger trial. PMID- 10805617 TI - "Preconditioning at a distance" in the isolated rabbit heart. AB - OBJECTIVE: Brief myocardial ischemia evokes a cardioprotective response, referred to as "ischemic preconditioning" (IP), that limits injury caused by a subsequent prolonged ischemic insult. The myocardial IP effect can be induced by ischemia of "distant" cardiac and noncardiac tissue, implicating the involvement of an as-yet unidentified humoral trigger. If a preconditioning hormone exists, the authors hypothesize that the IP effect should be transferable, via administration of coronary effluent, from a preconditioned donor heart to a virgin non preconditioned acceptor heart. METHODS: Isolated buffer-perfused rabbit hearts were assigned to one of four treatment groups in a donor/acceptor sequence. Donor hearts underwent either three IP cycles or a matched period of uninterrupted perfusion (control donors). Coronary perfusate collected from IP and control donor hearts was reoxygenated and transfused to virgin acceptor hearts. All hearts then underwent 30 minutes of global ischemia followed by 30 minutes of reperfusion. Left ventricular developed pressure (LVDP) (the authors' index of cardioprotection) was monitored throughout the protocol by a left ventricular (LV) balloon. RESULTS: In donor controls, LVDP assessed at 30 minutes post-reflow was restored to only 49 +/- 5% of baseline values. Recovery of LV function was significantly enhanced in both IP donor hearts (69 +/- 4%*) and IP acceptor hearts (70 +/- 6%*) vs donor controls (*p < 0.05), while, in acceptor controls, intermediate values of LVDP (62 +/- 7%) were obtained. CONCLUSION: The IP effect can be transferred between rabbit hearts, suggesting the presence of a humoral trigger signal for distant preconditioning. Isolating this hormone may have therapeutic and diagnostic implications in the management of acute myocardial ischemia. PMID- 10805618 TI - The effect of hypertension on the response to blood loss in a rodent model. AB - OBJECTIVE: Hypertensive patients having higher baseline peripheral resistance and sympathetic tone than normotensive patients may have aberrant responses to hemorrhage. In an attempt to further characterize this clinical observation, the authors compared the hemodynamic and metabolic responses to hemorrhage between spontaneously hypertensive rats (SHR) and normotensive rats (NTR). METHODS: Twenty adult rats (10 NTR and 10 SHR) were anesthetized with althesin via the intraperitoneal route. Femoral arteries were cannulated by cutdown. Twelve (6 SHR and 6 NTR) rats underwent controlled catheter hemorrhage of 25% of their total blood volumes. Eight rats (4 SHR and 4 NTR) served as nonhemorrhage controls. Mean arterial pressure (MAP) and base excess (BE) were measured prehemorrhage and then every 15 minutes for the next 120 minutes. Data were reported as mean +/- standard error of the mean (SEM). Group comparisons were analyzed by ANOVA with repeated values post-hoc by Bonferroni. Statistical significance was defined by an alpha = 0.05. RESULTS: Immediately after hemorrhage, the SHR group experienced a significantly (p < 0.001) greater drop in MAP of 70 +/- 4% in the SHR vs 40 +/- 6% in the NTR. Blood pressure in the NTR returned to control values 15 minutes after hemorrhage, but the SHR remained relatively hypotensive for the entire length of the experiment. Base excess in the SHR decreased significantly (p < 0.004) by 8.2 +/- 2 mmol/L from control values, as compared with no changes in BE for the NTR. CONCLUSIONS: The authors observed significant differences in the response to hemorrhage between hypertensive and normotensive rats. Hypertensive rats experienced a more profound hemorrhagic shock insult than normotensives for the same degree of blood loss. PMID- 10805619 TI - Clinically meaningful changes in quantitative measures of asthma severity. AB - OBJECTIVE: To determine minimum clinically meaningful improvements in peak expiratory flow rate (PEFR) and dyspnea visual analog score (VAS) in patients with acute asthma exacerbation. METHODS: Patients presenting to the emergency department (ED) with acute asthma exacerbation were eligible. The PEFR and VAS were assessed at presentation and after initial asthma therapy. During reassessment, subjects were asked to describe their asthma symptoms as "much better," "a little better," "no change," "a little worse," or "much worse." Correspondence between self-reported improvement and changes in PEFR and VAS was assessed. The "minimum clinically significant change" in either index was defined as the difference between pre- and posttreatment measures in subjects reporting their symptoms "a little better." RESULTS: One hundred fifty-six subjects were included. Asthma symptoms were "much better" in 99 (64%), "a little better" in 41 (26%), and "unimproved" (composed of patients describing symptoms as "no change," "a little worse," or "much worse") in 16 (10%). The mean VAS change among the "a little better" subjects was 2.2 cm (95% CI = 1.1 to 3.4), significantly greater than the -0.4 cm (95% CI = -2.1 to 1.4) change in the "unimproved" subjects. The mean change in percent predicted PEFR among the "a little better" subjects was 11.9 (95% CI = 7.3 to 16.1), not statistically different from the change of 6.1 (95% CI = 1.1 to 11.3) in the "no change" subjects. The "much better" group showed significantly greater changes in both measures than either of the other groups. A VAS change of > or =0.5 cm reliably discriminated between subjects with and without symptom improvement. CONCLUSIONS: Improvements in VAS of 2.2 cm and in predicted PEFR of about 12 percentage points are minimal clinically significant improvements during ED asthma therapy. The dyspnea VAS is valid in assessing symptomatic changes and may detect small subjective improvements better than the PEFR. PMID- 10805620 TI - Isolated intraperitoneal fluid on abdominal computed tomography in children with blunt trauma. AB - OBJECTIVE: To determine the frequency of isolated intraperitoneal fluid (IIF) on abdominal computed tomography (CT) in pediatric blunt trauma patients and the association between IIF and clinically identifiable intra-abdominal injuries (IAIs) in these patients. METHODS: The authors conducted a prospective observational study of consecutive children <16 years old with blunt torso trauma who underwent abdominal CT scanning while in the emergency department (ED). All patients were evaluated by a faculty emergency physician who documented the patient's physical examination. All CTs were interpreted by a single faculty radiologist masked to clinical data. The volume of intraperitoneal fluid was quantified (small, moderate, large) and the presence of organ injury visible on CT was noted. Patients were considered to have IIF if the CT demonstrated intraperitoneal fluid and no solid organ injury. Patients with IIF were followed through their hospitalizations or telephoned in one week if discharged home from the ED. RESULTS: Five hundred twenty-seven children with blunt trauma were enrolled into the study. The mean age (+/-SD) was 7.4 +/- 4.7 years, and the median pediatric trauma score was 10 (range -2 to 12). Eighty-eight patients (17%; 95% CI = 14% to 20%) had intraperitoneal fluid on CT scan and 42 (48%; 95% CI = 37% to 59%) of these patients had IIF. Of the 42 patients with IIF, five patients (all without abdominal tenderness and with a small amount of IIF on CT scan) were discharged to home from the ED and were well at telephone follow-up; the remaining 37 patients were hospitalized. Of the 42 patients with IIF, 7 patients (17%, 95% CI = 7 to 31%) had IAIs subsequently identified (all gastrointestinal injuries) during their evaluations. Six of the seven patients with IIF and subsequently identified IAIs had abdominal tenderness on examination in the ED. The remaining patient had a decreased level of consciousness. CONCLUSIONS: Isolated intraperitoneal fluid occurs in 8% of pediatric blunt trauma patients undergoing abdominal CT, and IAIs are subsequently identified in 17% of these patients. Patients with a small amount of IIF on CT who lack abdominal tenderness and have a normal level of consciousness are at low risk for subsequently identified IAIs. PMID- 10805621 TI - Clinical clearance of cervical spinal injuries by emergency nurses. AB - OBJECTIVES: To determine the interrater reliability between emergency nurses and emergency physicians on defined criteria for clinically clearing the cervical spine in blunt trauma patients. METHODS: Blunt trauma patients, 12 years or older, arriving with cervical spinal precautions were prospectively enrolled as a convenience sample. Each member of the emergency physician-nurse pair completed a questionnaire with regard to five criteria for clinically clearing the cervical spine for each patient. Interrater reliability was determined by calculating the kappa statistics for the individual and combined criteria. RESULTS: Physicians and nurses agreed on the presence or absence of the combined criteria in 175 of 211 patients (82.9%; kappa, 0.65). Agreements on individual criteria were as follows: 1) intoxication--203 patients (96.2%; kappa, 0.82); 2) altered consciousness--197 patients (93.4%; kappa, 0.60); 3) neck pain--185 patients (87.7%; kappa, 0.75); 4) distracting injury--160 patients (75.8%; kappa, 0.36); and 5) neurologic deficit--198 patients (93.8%; kappa, 0.45). If disagreements in which the physician would clinically clear the patient but the nurse would not were considered as agreements, then overall agreement would be 198 of 211 patients (93.8%; kappa, 0.88). On the assumption that nurses would assess patients prior to physicians, they would have cleared 35% of the patients before the physicians. However, they would have ordered 12% more radiographs and unsafely clinically cleared 5% of the patients. CONCLUSIONS: The interrater reliability for the combined cervical spinal injury criteria between emergency nurses and physicians was good to excellent. However, with the training given in this study, nurses would order more radiographs than physicians and would unsafely clinically clear cervical spines in some patients. PMID- 10805622 TI - The emergency department as a potential site for smoking cessation intervention: a randomized, controlled trial. AB - OBJECTIVE: To assess the effect of physician counseling and referral on smoking cessation rates and attendance at a smoking cessation program. METHODS: This was a prospective, randomized clinical trial set in a suburban, community teaching hospital emergency department (ED). During study hours, dedicated research associates enrolled consecutive, stable, oriented patients who were smokers. Eligible, consenting patients were randomized to one of two intervention groups. The control group received a two-page "Stop Smoking" pamphlet from the American Heart Association (AHA). Patients in the intervention group were given the AHA pamphlet along with pharmacologic information and standardized counseling by the attending emergency physician, including written and oral referral to a smoking cessation program. The primary outcome measures were telephone contact/attendance at the smoking cessation program by the intervention group and the rate of smoking cessation in both study groups at three months post-ED visit. Categorical data were analyzed by chi-square and Fisher's exact tests. Rank data were analyzed by Mann-Whitney tests and continuous data by t-tests. All tests were two tailed with alpha set at 0.05. RESULTS: One hundred fifty-two patients were enrolled; 78 were randomized to the intervention group. Nearly 70% of patients (103) were available for telephone follow-up. The study groups were statistically similar with regard to baseline demographic characteristics and the prevalence of moderate or severe nicotine addiction. None of the patients (0%) in the intervention group contacted or attended the smoking cessation program during the study period (95% CI = 0-4%). The percentages of patients who stopped smoking after three months were similar in the two groups [10.4% (5/48) control vs 10.9% (6/55) intervention; p = 1]. CONCLUSION: The authors found no difference in the smoking cessation rates between ED patients who received written material and those who were counseled by emergency physicians. Referral of patients who smoked to a cessation program was unsuccessful. PMID- 10805623 TI - Vaccination of emergency department patients at high risk for influenza. AB - OBJECTIVE: To determine the proportion of Canadian emergency department (ED) patients who are at risk for increased morbidity from influenza but were not vaccinated and to determine emergency physicians' (EPs') willingness to screen for and prescribe influenza vaccination. METHODS: The authors surveyed a convenience sample of patients presenting during a one-week period at each of four EDs in Winnipeg, Manitoba, Canada, after the end of the seasonal period for vaccination. They also surveyed all full-time EPs in Winnipeg. RESULTS: Fifty three percent of emergency patients at risk for increased morbidity from influenza had not been vaccinated and 59.3% of them were willing to be vaccinated during an emergency visit. This represents 31.6% (+/-3.1%) of all high-risk patients and 15% of all emergency patients. High-risk patients who did not have a regular physician were less likely to have been vaccinated (OR 0.165, p = 0.018). Most EPs rarely or never offer influenza vaccination (30% and 57%, respectively). Seventy-six percent of them were willing to prescribe vaccination. CONCLUSION: Many ED patients are at risk for increased morbidity from influenza and have not been vaccinated. The majority of them are willing to be vaccinated during an emergency visit and the majority of EPs are willing to prescribe vaccination. Emergency department vaccination for influenza should be considered as a strategy to increase vaccination among high-risk groups. PMID- 10805624 TI - Proposed curriculum for an "observational" international emergency medicine fellowship program. AB - This article presents information on considerations involved in setting up and conducting fellowship training programs in emergency medicine (EM) for physicians from other countries. General goals for these programs are to assist in providing physicians from other countries with the knowledge and skills needed to further develop EM in their home countries. The authors report their opinions, based on their cumulative extensive experiences, on the necessary and optional structural elements to consider for international EM fellowship programs. Because of U.S. medical licensing restrictions, much of the proposed programs' content would be "observational" rather than involving direct "hands-on" clinical EM training. Due to the very recent initiation of these programs in the United States, there has not yet been reported any scientific evaluation of their structure or efficacy. International EM fellowship programs involving mainly observational EM experience can serve as one method to assist in EM development in other countries. Future studies should assess the impact and efficacy of these programs. PMID- 10805625 TI - Clinicopathological conference: a case of ascending numbness. PMID- 10805626 TI - Leprosy in the emergency department. AB - OBJECTIVES: Los Angeles County-University of Southern California Medical Center, like many large urban hospitals, has a large immigrant population from regions of the world where leprosy is endemic. Emergency physicians (EPs) in these settings can expect to encounter leprosy patients. This study reviewed the emergency department (ED) course of patients with confirmed leprosy in an attempt to describe the most common presenting patterns so that future cases can be more easily recognized. METHODS: This was a retrospective chart review of all patients followed in the Hansen's disease clinic. Demographics, leprosy type, clinical presentations to the ED, and medications were recorded. RESULTS: Of the total number of patients (415), most were of Mexican (52%), Filipino (15%), Vietnamese (14%), and Chinese (5%) origin. Leprosy was classified as lepromatous (56%), borderline (40%), and tuberculoid (4%). There were a total of 118 ED visits by 74 patients. The mean age was 46 years, with 51% male and 49% female. Dermatologic (68%), neurologic (23%), and ophthalmologic (9%) complaints were the most common reasons for ED presentation related to leprosy. The EP did not elicit a history of leprosy in 34% of those patients followed in the leprosy clinic. The ED diagnosis of leprosy was made in 3 of 15 (20%) undiagnosed cases. Of the 63 patients prescribed medications in the leprosy clinic at the time of their ED visits, 22 (35%) ED charts did not report leprosy drugs. CONCLUSION: Patients with leprosy present to U.S. EDs, and new cases can be identified. Early recognition is important given leprosy's devastating consequences, major drug side effects of medications used for treatment, and improved prognosis with multidrug therapy. A history of leprosy and associated medications are often not documented in the ED chart, which may reflect a continued fear of stigmatization among these patients. PMID- 10805627 TI - Mission-based management: implications for emergency medicine. PMID- 10805628 TI - The aerial palaces of decision analysis redux. PMID- 10805629 TI - So what's a little free fluid? PMID- 10805630 TI - Diagnostic evaluation of patients with blunt abdominal trauma: a decision analysis. AB - OBJECTIVES: Using decision analysis, to compare the expected utility (EU) of diagnostic peritoneal lavage (DPL), computed tomography (CT), and ultrasonography (US) to determine the optimal modality for the evaluation of blunt abdominal trauma (BAT) in hemodynamically stable adults. METHODS: Data points for the decision analysis were obtained from three sources: 1) prevalence of BAT and the sensitivity and specificity of each diagnostic modality were determined through a criteria-based review of the literature; 2) rate of BAT necessitating immediate intervention, perioperative complication rate, and operative mortality rate were calculated using data from the authors' institution's trauma registry; and 3) outcome utilities were determined by telephone survey of adults in a random sample of households in the region. The decision tree was constructed and evaluated in standard fashion. For each diagnostic modality, the authors calculated the EU using the minimum, mean, and maximum sensitivity and specificity across a range of prevalence. Mean outcome utilities were used for each branch of the tree when calculating the EU. RESULTS: The EU of CT was consistently lower than the EUs of DPL and US at all levels of prevalence. However, the rank order of the EUs of US and DPL varied with the prevalence of BAT. When the prevalence was <30%, the EU of US was higher than that for DPL. When the prevalence was 30-40%, the EUs were similar. When the prevalence was >40%, the EU of US was less than that of DPL. CONCLUSIONS: Among institutions operating under constraints similar to those used in this model, the optimal diagnostic modality for the evaluation of BAT can be determined based on the sensitivity and specificity of the modality at their institution and the prevalence of BAT in their patient population. PMID- 10805631 TI - Research fundamentals: selection and development of clinical outcome measures. AB - Clinical outcomes measure patient health or well-being. The choice of an outcome measure for use in a clinical trial or study is complex. It is even more difficult when no appropriate outcome measure exists, necessitating the development of a novel one. The ideal clinical outcome should be credible, comprehensive, sensitive to change, accurate, biologically sensible, and feasible. This paper describes the attributes of clinical outcomes and illustrates how to develop novel outcomes, using as an example the authors' experience in developing a wound cosmesis outcome measure. PMID- 10805632 TI - Out-of-hospital management of stroke: surveying local practice with implication for change. PMID- 10805633 TI - Nebulized or sprayed lidocaine as anesthesia for nasogastric intubations. PMID- 10805634 TI - Characteristics of the emergency medicine standardized letter of recommendation. PMID- 10805635 TI - Comments on "Clinical decision making: an emergency medicine perspective". PMID- 10805636 TI - Comments on "Clinical decision making: an emergency medicine perspective". PMID- 10805637 TI - An alternative manpower source for clinical research. PMID- 10805638 TI - Comments on "Emergency department observation of poisoned patients: how long is necessary?". PMID- 10805639 TI - The Logic and Realism of the Hypothesis of Exploitation Ecosystems. AB - Hypotheses on trophic dynamics in terrestrial ecosystems fall into two major categories: those in which plants are assumed to be invulnerable to their consumers and those in which the build-up of plant biomass is assumed to require top-down control of folivores. The hypothesis of exploitation ecosystems (EEH) belongs to the latter category and focuses particularly on the consequences of the high energetic costs of maintenance of endotherms. Carnivorous endotherms require relatively high prey densities in order to break even. Moreover, they are dependent on folivorous prey during the limiting season, at least at high latitudes. The endotherm branch of the grazing web is thus predicted to collapse from three-link trophic dynamics (carnivores -> folivores -> plants -> inorganic resources) to two-link dynamics (folivores -> plants -> inorganic resources) along gradients of decreasing primary productivity. Consequently, the vegetation of cold and unproductive areas is predicted to be under intense winter grazing pressure, which prevents the accumulation of aboveground plant biomass and excludes erect woody plants. In the most extreme habitats (e.g., polar deserts and their high alpine counterparts), even folivorous endotherms are predicted to be absent, and the scanty vegetation is predicted to be structured by preemptive competition. Within temperature-determined productivity gradients, EEH is corroborated by biomass patterns, by patterns in the structure and dynamics of carnivore, folivore, and plant communities, and by experimental results. The general idea of top-down trophic dynamics is supported for other autotroph-based systems, too, but the relevance and sufficiency of the energy constraint in explaining patterns in trophic dynamics appears to be variable. Moreover, critical empirical evidence for or against the capacity of folivorous insects to regulate plant biomass has not yet been obtained. Another open question is the ability of boreal and temperate browsers, evolved in productive environments with intense predation pressure and abundance of forage, to prevent the regeneration of the least palatable tree species. There are, thus, many open questions waiting to be answered and many exciting experiments waiting to be conducted. PMID- 10805640 TI - Bet-Hedging Diapause Strategies in Stochastic Environments. AB - In many insect species, adult emergence spreads over several years because of the existence of prolonged diapause in certain individuals. From stochastic models, we show that diversified bet-hedging strategies (mixed strategies with emergence after 1 or 2 yr) are more fit than simple diapause strategy (emergence after 1 yr) or fixed prolonged diapause strategy (emergence after 2 yr) in isolated chestnut weevil populations. This conclusion applies to a large range of survival rates in prolonged diapause and is insensitive to initial conditions, magnitude of temporal autocorrelation, distribution of demographic parameters, and quoted values of population size limitation. However, the shape of the fitness distribution as a function of prolonged diapause frequency changes greatly in the absence of population size limitation. Whatever the survival rate during prolonged diapause, we find that there is no genotypic advantage to extending diapause for all chestnut weevil larvae to more than 1 yr. Our models predict selection of bet-hedging strategies over a large range of prolonged diapause frequencies. This result is consistent with the existence of several mixed strategies in a population. Emergences after 3 yr are not crucial for selection or for the dynamics of mixed strategies in the chestnut weevil. PMID- 10805641 TI - Effect of Herbivory and Plant Species Replacement on Primary Production. AB - Grazing optimization occurs when herbivory increases primary production at low grazing intensities. In the case of simple plant-herbivore interactions, such an effect can result from recycling of a limiting nutrient. However, in more complex cases, herbivory can also lead to species replacement in plant communities, which in turn alters how primary production is affected by herbivory. Here we explore this issue using a model of a limiting nutrient cycle in an ecosystem with two plant species. We show that two major plant traits determine primary production at equilibrium: plant recycling efficiency (i.e., the fraction of the plant nutrient stock that stays within the ecosystem until it is returned to the nutrient pool in mineral form) and plant ability to deplete the soil mineral nutrient pool through consumption of this resource. In cases where sufficient time has occurred, grazing optimization requires that herbivory improve nutrient conservation in the system sufficiently. This condition sets a minimum threshold for herbivore nutrient recycling efficiency, the fraction of nutrient consumed by herbivores that is recycled within the ecosystem to the mineral nutrient pool. This threshold changes with plant community composition and herbivore preference and is, therefore, strongly affected by plant species replacement. The quantitative effects of these processes on grazing optimization are determined by both the recycling efficiencies and depletion abilities of the plant species. However, grazing optimization remains qualitatively possible even with plant species replacement. PMID- 10805642 TI - Comparative Phylogeography of Mesoamerican Highland Rodents: Concerted versus Independent Response to Past Climatic Fluctuations. AB - The phylogeography of Sumichrast's harvest mouse (Reithrodontomys sumichrasti) was examined through maximum-likelihood and parsimony analyses of 1,130 bp of mitochondrial Cytochrome b sequence data from 43 individuals. The phylogeography of this Middle American highland forest-dwelling species was compared to that previously published for the codistributed Aztec deer mouse complex (Peromyscus aztecus/Peromyscus hylocetes complex) in order to test competing hypotheses of concerted versus independent responses of codistributed forms to past climatic fluctuations. Qualitatively, there were strong similarities in the phylogeographic patterns of the two groups, yet there were also areas of incongruence. Likelihood-ratio tests (Kishino-Hasegawa-Templeton and parametric bootstrap tests) indicated that this incongruence is significant and cannot be attributed simply to uncertainty in phylogenetic estimation, thereby falsifying the concerted-response hypothesis. Conversely, tree-reconciliation analysis of the area relationships inferred for each group separately indicated that there has been a significant history of covicariance between the two groups, falsifying the independent-response hypothesis. It appears that codistributed taxa in the geologically complex highlands of Mesoamerica share more common biogeographical history than can be accounted for by the independent-response hypothesis yet have not responded to past climatic fluctuations in the lock-step fashion predicted by the concerted-response hypothesis. PMID- 10805643 TI - Competition and the Effect of Spatial Resource Heterogeneity on Evolutionary Diversification. AB - A model is presented to explore how the form of selection arising from competition for resources is affected by spatial resource heterogeneity. The model consists of a single species occupying two patches connected by migration, where the two patches can differ in the type of resources that they contain. The main goal is to determine the conditions under which competition for resources results in disruptive selection (i.e., selection favoring a polymorphism) since it is this form of selection that will give rise to the evolutionary diversification of resource exploitation strategies. In particular, comparing the conditions giving rise to disruptive selection when the two patches are identical to the conditions when they contain different resources reveals the effect of spatial resource heterogeneity. Results show that when the patches are identical, the conditions giving rise to disruptive selection are identical to those that give rise to character displacement in previous models. When the patches are different, the conditions giving rise to disruptive selection can be either more or less stringent depending upon demographic parameters such as the intrinsic rate of increase and the migration rate. Surprisingly, spatial resource heterogeneity can actually make forms of evolutionary diversification such as character displacement less likely. It is also found that results are dependent on how the resource exploitation strategies and the spatial resource heterogeneity affect the population dynamics. One robust conclusion however, is that spatial resource heterogeneity always has a disruptive effect when the migration rate between patches is low. PMID- 10805644 TI - Competition and the Effect of Spatial Resource Heterogeneity on Evolutionary Diversification. AB - A model is presented to explore how the form of selection arising from competition for resources is affected by spatial resource heterogeneity. The model consists of a single species occupying two patches connected by migration, where the two patches can differ in the type of resources that they contain. The main goal is to determine the conditions under which competition for resources results in disruptive selection (i.e., selection favoring a polymorphism) since it is this form of selection that will give rise to the evolutionary diversification of resource exploitation strategies. In particular, comparing the conditions giving rise to disruptive selection when the two patches are identical to the conditions when they contain different resources reveals the effect of spatial resource heterogeneity. Results show that when the patches are identical, the conditions giving rise to disruptive selection are identical to those that give rise to character displacement in previous models. When the patches are different, the conditions giving rise to disruptive selection can be either more or less stringent depending upon demographic parameters such as the intrinsic rate of increase and the migration rate. Surprisingly, spatial resource heterogeneity can actually make forms of evolutionary diversification such as character displacement less likely. It is also found that results are dependent on how the resource exploitation strategies and the spatial resource heterogeneity affect the population dynamics. One robust conclusion however, is that spatial resource heterogeneity always has a disruptive effect when the migration rate between patches is low. PMID- 10805645 TI - Ectotherms, Temperature, and Trade-offs: Size and Number of Eggs in a Carabid Beetle. AB - We studied the allocation of total egg mass to size and number in the carabid beetle Notiophilus biguttatus F. at several temperature and day length regimes. Eggs increase in number and decrease in size with increasing (constant) temperature. Day length interacts with temperature: at short day the effect of temperature on size and number of eggs is weaker than at long day. In diurnally fluctuating temperature regimes, egg size is affected disproportionately by the high temperature period. All treatments, however, are similar in affecting number and size of eggs in an opposite direction. Consequently, egg size is explained to a high degree by egg production rate. The relationship between size and number of eggs among treatments is furthermore characterized by a decrease in egg size with an increase in total egg mass production. Within treatments, rate of egg production and egg size are negatively correlated among females in the low temperature groups but not in the high-temperature groups; the correlations among females are also characterized by a decrease in egg size, with an increase in total egg mass production. Hence, possible trade-offs between size and number of eggs are masked by phenotypic variation in reproductive effort. The observations enable us to propose a simple conceptual model that explains the within-treatment correlation by the same causal factor as the negative relationship among treatment means. PMID- 10805646 TI - Population Structure Influences Sex Ratio Evolution in a Gynodioecious Plant. PMID- 10805647 TI - No Behavioral Control over Mating Frequency in Queen Honey Bees (Apis mellifera L.): Implications for the Evolution of Extreme Polyandry. PMID- 10805649 TI - Multifunctional neuron CC6 in Aplysia exerts actions opposite to those of multifunctional neuron CC5. AB - The controls of somatic and autonomic functions often appear to be organized into antagonistic systems. This issue was explored in the bilaterally paired C cluster neuron, CC6, which was found to have properties that suggested that it might function antagonistically to the previously identified multiaction neuron, CC5. Similar to CC5, CC6 is an interganglionic neuron that sends its sole axon to the ipsilateral and contralateral pedal and pleural ganglia. Synaptic inputs to CC6 were opposite to those of CC5. For example, CC6 receives inhibitory inputs from mechanical touch to the lips and tentacles and is excited by firing of C-PR, a neuron involved in the control of a head extension response. Also during rhythmic buccal mass movements CC6 receives synaptic inputs that are out of phase with those received by CC5. CC6 is inhibited during a fictive locomotor program, whereas CC5 is excited, but unlike CC5, the inputs to CC6 are not rhythmic. CC6 has extensive mono- and polysynaptic outputs to many identified and unidentified neurons located in various central ganglia. Firing of CC6 evoked ipsilateral contraction of the transverse muscles of the neck, whereas CC5 contracts longitudinal neck muscles. CC6 monosynaptically inhibits the pedal artery shortener neuron, whereas CC5 monosynaptically excites the pedal artery shortener neuron. Specific motor neurons in the pedal ganglion receive synaptic inputs of opposite sign from CC5 and CC6. Although the inputs and most of the effects of CC6 were opposite to those of CC5, both cells were found to produce polysynaptic excitation of the abdominal ganglion neuron RBhe, a cell whose activity excites the heart. CC5 and CC6 appear to be multifunctional neurons that form an antagonist pair. PMID- 10805651 TI - Presynaptic and interactive peptidergic modulation of reticulospinal synaptic inputs in the lamprey. AB - The modulatory effects of neuropeptides on descending inputs to the spinal cord have been examined by making paired recordings from reticulospinal axons and spinal neurons in the lamprey. Four peptides were examined; peptide YY (PYY) and cholecystokinin (CCK), which are contained in brain stem reticulospinal neurons, and calcitonin-gene-related peptide (CGRP) and neuropeptide Y (NPY), which are contained in primary afferents and sensory interneurons, respectively. Each of the peptides reduced the amplitude of monosynaptic reticulospinal-evoked excitatory postsynaptic potentials (EPSPs). The modulation appeared to be presynaptic, because postsynaptic input resistance and membrane potential, the amplitude of the electrical component of the EPSP, postsynaptic responses to glutamate, and spontaneous miniature EPSP amplitudes were unaffected. In addition, none of the peptides affected the pattern of N-methyl-D-aspartate (NMDA)-evoked locomotor activity in the isolated spinal cord. Potential interactions between the peptides were also examined. The "brain stem peptides" CCK and PYY had additive inhibitory effects on reticulospinal inputs, as did the "sensory peptides" CGRP and NPY. Brain stem peptides also had additive inhibitory effects when applied with sensory peptides. However, sensory peptides increased or failed to affect the amplitude of reticulospinal inputs in the presence of the brain stem peptides. These interactive effects also appear to be mediated presynaptically. The functional consequence of the peptidergic modulation was investigated by examining spinal ventral root responses elicited by brain stem stimulation. CCK and CGRP both reduced ventral root responses, although in interaction both increased the response. These results thus suggest that neuropeptides presynaptically influence the descending activation of spinal locomotor networks, and that they can have additive or novel interactive effects depending on the peptides examined and the order of their application. PMID- 10805650 TI - Horizontal vestibuloocular reflex evoked by high-acceleration rotations in the squirrel monkey. III. Responses after labyrinthectomy. AB - The horizontal angular vestibuloocular reflex (VOR) evoked by high-frequency, high-acceleration rotations was studied in four squirrel monkeys after unilateral labyrinthectomy. Spontaneous nystagmus was measured at the beginning and end of each testing session. During the period that animals were kept in darkness (4 days), the nystagmus at each of these times measured approximately 20 degrees /s. Within 18-24 h after return to the light, the nystagmus (measured in darkness) decreased to 2.8 +/- 1.5 degrees /s (mean +/- SD) when recorded at the beginning but was 20.3 +/- 3.9 degrees /s at the end of the testing session. The latency of the VOR measured from responses to steps of acceleration (3,000 degrees /s(2) reaching a velocity of 150 degrees /s) was 8.4 +/- 0.3 ms for responses to ipsilesional rotations and 7.7 +/- 0.4 ms for contralesional rotations. During the period that animals were kept in darkness after the labyrinthectomy, the gain of the VOR measured during the steps of acceleration was 0.67 +/- 0.12 for contralesional rotations and 0.39 +/- 0.04 for ipsilesional rotations. Within 18 24 h after return to light, the VOR gain for contralesional rotations increased to 0.87 +/- 0.08, whereas there was only a slight increase for ipsilesional rotations to 0.41 +/- 0. 06. A symmetrical increase in the gain measured at the plateau of head velocity was noted after the animals were returned to light. The VOR evoked by sinusoidal rotations of 2-15 Hz, +/-20 degrees /s, showed a better recovery of gain at lower (2-4 Hz) than at higher (6-15 Hz) frequencies. At 0.5 Hz, gain decreased symmetrically when the peak amplitude was increased from 20 to 100 degrees /s. At 10 Hz, gain was decreased for ipsilesional half-cycles and increased for contralesional half-cycles when velocity was raised from 20 to 50 degrees /s. A model incorporating linear and nonlinear pathways was used to simulate the data. Selective increases in the gain for the linear pathway accounted for the recovery in VOR gain for responses at the velocity plateau of the steps of acceleration and for the sinusoidal rotations at lower peak velocities. The increase in gain for contralesional responses to steps of acceleration and sinusoidal rotations at higher frequencies and velocities was due to an increase in the contribution of the nonlinear pathway. This pathway was driven into cutoff and therefore did not affect responses for rotations toward the lesioned side. PMID- 10805652 TI - Effects of acute and chronic midthoracic spinal cord injury on neural circuits for male sexual function. II. Descending pathways. AB - In normal animals, microstimulation of the medullary reticular formation (MRF) has two effects on efferent neurons in the motor branch of the pudendal nerve (PudM). MRF microstimulation depresses motoneuron reflex discharges (RD) elicited by dorsal nerve of the penis (DNP) stimulation and produces long latency sympathetic fiber responses (SFR). The midthoracic spinal location of these descending MRF-PudM projections was studied electrophysiologically using a variety of acute and chronic lesions. Chronic lesions, in 27 mature male rats, included dorsal (DHx) or lateral (LHx) hemisections or moderate/severe contusions (Cx) at spinal level T(8). Behavioral data (sexual reflex latency, bladder voiding) obtained throughout the recovery period revealed a significant impairment of urogenital function for the DHx and severe Cx groups of animals. Microstimulation-induced PudM-RDs and PudM-SFRs, obtained in terminal electrophysiological experiments 30 days postinjury in the same 27 rats (urethan anesthetized), were tested for a combined total of 1,404 bilateral MRF sites. PudM-RD was obtained for LHx and moderate Cx groups of animals but not for DHx or severe Cx groups. PudM-SFRs were obtained for LHx, DHx (although significantly weakened) and moderate Cx groups but not for those having received either an over DHx or a severe Cx injury. PudM responses also were tested for 6 MRF sites in six intact control rats both before and after various select acute spinal cord lesions. For MRF sites producing a robust PudM-RD and PudM-SFR, acute bilateral lesions confined to the dorsolateral quadrant (DLQ) eliminated the PudM-RD but failed to eliminate PudM-SFRs. A deeper lesion encompassing additional white matter located dorsally in the ventrolateral quadrant (VLQ) was necessary to eliminate PudM-SFRs. Overall, these electrophysiological results provide evidence for descending projections conveying information between MRF and the lower thoracic/lumbosacral male urogenital circuitry within the DLQ and the dorsal-most aspect of VLQ at the midthoracic level of spinal cord. The alterations of supraspinal projections observed after chronic injury are likely of important clinical significance for functional recovery in cases of clinically incomplete spinal cord injury at midthoracic spinal cord. PMID- 10805653 TI - Activation of presynaptic group III metabotropic receptors enhances glutamate release in rat entorhinal cortex. AB - The role of group III metabotropic glutamate receptors (mGluRs) in modulating excitatory synaptic transmission was investigated in the rat entorhinal cortex (EC) in vitro. AMPA receptor-mediated excitatory postsynaptic currents (EPSCs) were recorded in the whole cell configuration of the patch-clamp technique from visually identified neurons in layers V and II. In layer V, bath application of the specific group III mGluR agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4, 500 microM) resulted in a marked facilitation of both spontaneous and activity independent "miniature" (s/mEPSC) event frequency. The facilitatory effect of L AP4 (100 microM) on sEPSC frequency prevailed in the presence of DL-2-amino-5 phosphonopentanoic acid (100 microM) but was abolished by the group III antagonist (RS)-cyclopropyl-4-phosphonophenylglycine (20 microM). These data confirmed that group III mGluRs, and not N-methyl-D-aspartate (NMDA) receptors were involved in the response to L-AP4. Bath application of the specific mGluR4a agonist (1S,3R,4S)-1-aminocyclopentane-1,2, 4-tricarboxylic acid (20 microM) also had a facilitatory effect on sEPSC frequency, suggesting involvement of mGluR4a. In layer II neurons, L-AP4 caused a reduction in sEPSC frequency but did not affect mEPSCs recorded in the presence of tetrodotoxin. These findings suggest that a group III mGluR with mGluR4a-like pharmacology is involved in modulating synaptic transmission in layer V cells of the EC. The effect on mEPSCs suggests that this receptor is located presynaptically and that its activation results in a direct facilitation of glutamate release. This novel facilitatory effect is specific to layer V and, to our knowledge, is the first report of a direct facilitatory action of group III mGluRs on synaptic transmission. In layer II, L AP4 had an inhibitory effect on glutamate release similar to that reported in other brain regions. PMID- 10805654 TI - Sweet taste transduction in hamster: role of protein kinases. AB - Two different second-messenger pathways have been implicated in sweet taste transduction: sugars produce cyclic AMP (cAMP), whereas synthetic sweeteners stimulate production of inositol 1,4, 5-tris-phosphate (IP(3)) and diacylglycerol (DAG). Both sugars and sweeteners depolarize taste cells by blocking the same resting K(+) conductance, but the intermediate steps in the transduction pathways have not been examined. In this study, the loose-patch recording technique was used to examine the role of protein kinases and other downstream regulatory proteins in the two sweet transduction pathways. Bursts of action currents were elicited from approximately 35% of fungiform taste buds in response to sucrose (200 mM) or NC-00274-01 (NC-01, 200 microM), a synthetic sweetener. To determine whether protein kinase C (PKC) plays a role in sweet transduction, taste buds were stimulated with the PKC activator PDBu (10 microM). In all sweet-responsive taste buds tested (n = 11), PDBu elicited burst of action currents. In contrast, PDBu elicited responses in only 4 of 19 sweet-unresponsive taste buds. Inhibition of PKC by bisindolylmaleimide I (0.15 microM) resulted in inhibition of the NC-01 response by approximately 75%, whereas the response to sucrose either increased or remained unchanged. These data suggest that activation of PKC is required for the transduction of synthetic sweeteners. To determine whether protein kinase A (PKA) is required for the transduction of sugars, sweet responses were examined in the presence of the membrane-permeant PKA inhibitor H-89 (10 and 19 microM). Surprisingly, H-89 did not decrease responses to either sucrose or NC-01. Instead, responses to both compounds were increased in the presence of the inhibitor. These data suggest that PKA is not required for the transduction of sugars, but may play a modulatory role in both pathways, such as adaptation of the response. We also examined whether Ca(2+)-calmodulin dependent cAMP phosphodiesterase (CaM-PDE) plays a role in sweet taste transduction, by examining responses to sucrose and synthetic sweeteners in the presence of the CaM-PDE inhibitor W-7 (100 microM). Inhibition resulted in an increase in the response to sucrose, whereas the response to NC-01 remained unchanged. These data suggest that the pathways for sugars and sweeteners are negatively coupled; the Ca(2+) that is released from intracellular stores during stimulation with synthetic sweeteners may inhibit the response to sucrose by activation of CaM PDE. PMID- 10805655 TI - Novel mechanism of inhibition by the P2 receptor antagonist PPADS of ATP activated current in dorsal root ganglion neurons. AB - The antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) has been proposed to selectively antagonize the actions of ATP at P2X receptors. Whole cell patch-clamp recording techniques therefore were used to characterize PPADS inhibition of ATP-activated current in bullfrog dorsal root ganglion (DRG) neurons. PPADS, 0.5-10 microM, inhibited ATP-activated current in a concentration dependent manner with an IC(50) of 2.5 +/- 0.03 microM. PPADS produced a gradual decline of ATP-activated current to a steady state, but this was not an indication of use dependence as the gradual declining component could be eliminated by exposure to PPADS before ATP application. In addition, ATP activated current recovered completely from inhibition by PPADS in the absence of agonist. The slow onset of inhibition by PPADS was not apparently due to an action at an intracellular site as inclusion of 10 microM PPADS in the recording pipette neither affected the ATP response nor did it alter inhibition of the ATP response when 2.5 microM PPADS was applied externally. PPADS, 2.5 microM, decreased the maximal response to ATP by 51% without changing its EC(50). PPADS inhibition of ATP-activated current was independent of membrane potential between -80 and +40 mV and did not involve a shift in the reversal potential of the current. The magnitude of PPADS inhibition of ATP-activated current was dependent on the duration of the prior exposure to PPADS. The time constants of both onset and offset of PPADS inhibition of ATP-activated current did not differ significantly with changes in ATP concentration from 1 to 5 microM. Recovery of ATP-activated current from PPADS inhibition also exhibited a slow phase that was not accelerated by the presence of agonist and was dependent on the concentration of PPADS. The apparent dissociation rate of PPADS from unliganded ATP-gated ion channels was much greater than the rate of the slow phase of recovery of ATP activated current from PPADS inhibition. The results suggest that PPADS can inhibit P2X receptor function in a complex noncompetitive manner. PPADS produces a long-lasting inhibition that does not appear to result from open channel block but rather from an action at an allosteric site apparently accessible from the extracellular environment that involves a greatly reduced rate of dissociation from liganded versus unliganded ATP-gated ion channels. PMID- 10805656 TI - Short-term potentiation of miniature excitatory synaptic currents causes excitation of supraoptic neurons. AB - Magnocellular neurons (MCNs) of the hypothalamic supraoptic nucleus (SON) secrete vasopressin and oxytocin. With the use of whole-cell and nystatin-perforated patch recordings of MCNs in current- and voltage-clamp modes, we show that high frequency stimulation (HFS, 10-200 Hz) of excitatory afferents induces increases in the frequency and amplitude of 2,3-dioxo-6-nitro-1,2,3, 4 tetrahydrobenzo(f)quinoxaline-7-sulfonamide (NBQX)-sensitive miniature excitatory postsynaptic currents (mEPSCs) lasting up to 20 min. This synaptic enhancement, referred to as short-term potentiation (STP), could be induced repeatedly; required tetrodotoxin (TTX)-dependent action potentials to initiate, but not to maintain; and was independent of postsynaptic membrane potential, N-methyl-D aspartate (NMDA) receptors, or retrograde neurohypophyseal neuropeptide release. STP was not accompanied by changes in the conductance of the MCNs or in the responsiveness of the postsynaptic non-NMDA receptors, as revealed by brief application of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate. mEPSCs showed similar rise times before and after HFS and analysis of amplitude distributions of mEPSCs revealed one or more peaks pre-HFS and the appearance of additional peaks post-HFS, which were equidistant from the first peak. STP of mEPSCs was not associated with enhanced evoked responses, but was associated with an NBQX-sensitive increase in spontaneous activity of MCNs. Thus we have identified a particularly long-lasting potentiation of excitatory synapses in the SON, which has a presynaptic locus, is dissociated from changes in evoked release, and which regulates postsynaptic cell excitability. PMID- 10805657 TI - Ca(2+) channels involved in the generation of the slow afterhyperpolarization in cultured rat hippocampal pyramidal neurons. AB - The advantages of using isolated cells have led us to develop short-term cultures of hippocampal pyramidal cells, which retain many of the properties of cells in acute preparations and in particular the ability to generate afterhyperpolarizations after a train of action potentials. Using perforated patch recordings, both medium and slow afterhyperpolarization currents (mI(AHP) and sI(AHP), respectively) could be obtained from pyramidal cells that were cultured for 8-15 days. The sI(AHP) demonstrated the kinetics and pharmacologic characteristics reported for pyramidal cells in slices. In addition to confirming the insensitivity to 100 nM apamin and 1 mM TEA, we have shown that the sI(AHP) is also insensitive to 100 nM charybdotoxin but is inhibited by 100 microM D tubocurarine. Concentrations of nifedipine (10 microM) and nimodipine (3 microM) that maximally inhibit L-type calcium channels reduced the sI(AHP) by 30 and 50%, respectively. However, higher concentrations of nimodipine (10 microM) abolished the sI(AHP), which can be partially explained by an effect on action potentials. Both nifedipine and nimodipine at maximal concentrations were found to reduce the HVA calcium current in freshly dissociated neurons to the same extent. The N-type calcium channel inhibitor, omega-conotoxin GVIA (100 nM), irreversibly inhibited the sI(AHP) by 37%. Together, omega-conotoxin (100 nM) and nifedipine (10 microM) inhibited the sI(AHP) by 70%. 10 microM ryanodine also reduced the sI(AHP) by 30%, suggesting a role for calcium-induced calcium release. It is concluded that activation of the sI(AHP) in cultured hippocampal pyramidal cells is mediated by a rise in intracellular calcium involving multiple pathways and not just entry via L-type calcium channels. PMID- 10805658 TI - Properties and role of I(h) in the pacing of subthreshold oscillations in entorhinal cortex layer II neurons. AB - Various subsets of brain neurons express a hyperpolarization-activated inward current (I(h)) that has been shown to be instrumental in pacing oscillatory activity at both a single-cell and a network level. A characteristic feature of the stellate cells (SCs) of entorhinal cortex (EC) layer II, those neurons giving rise to the main component of the perforant path input to the hippocampal formation, is their ability to generate persistent, Na(+)-dependent rhythmic subthreshold membrane potential oscillations, which are thought to be instrumental in implementing theta rhythmicity in the entorhinal-hippocampal network. The SCs also display a robust time-dependent inward rectification in the hyperpolarizing direction that may contribute to the generation of these oscillations. We performed whole cell recordings of SCs in in vitro slices to investigate the specific biophysical and pharmacological properties of the current underlying this inward rectification and to clarify its potential role in the genesis of the subthreshold oscillations. In voltage-clamp conditions, hyperpolarizing voltage steps evoked a slow, noninactivating inward current, which also deactivated slowly on depolarization. This current was identified as I(h) because it was resistant to extracellular Ba(2+), sensitive to Cs(+), completely and selectively abolished by ZD7288, and carried by both Na(+) and K(+) ions. I(h) in the SCs had an activation threshold and reversal potential at approximately -45 and -20 mV, respectively. Its half-activation voltage was -77 mV. Importantly, bath perfusion with ZD7288, but not Ba(2+), gradually and completely abolished the subthreshold oscillations, thus directly implicating I(h) in their generation. Using experimentally derived biophysical parameters for I(h) and the low-threshold persistent Na(+) current (I(NaP)) present in the SCs, a simplified model of these neurons was constructed and their subthreshold electroresponsiveness simulated. This indicated that the interplay between I(NaP) and I(h) can sustain persistent subthreshold oscillations in SCs. I(NaP) and I(h) operate in a "push-pull" fashion where the delay in the activation/deactivation of I(h) gives rise to the oscillatory process. PMID- 10805659 TI - Selectivity for the shape, size, and orientation of objects for grasping in neurons of monkey parietal area AIP. AB - In this study, we mainly investigated the visual selectivity of hand-manipulation related neurons in the anterior intraparietal area (area AIP) while the animal was grasping or fixating on three-dimensional (3D) objects of different geometric shapes, sizes, and orientations. We studied the activity of 132 task-related neurons during the hand-manipulation tasks in the light and in the dark, as well as during object fixation. Seventy-seven percent (101/132) of the hand manipulation-related neurons were visually responsive, showing either lesser activity during manipulation in the dark than during that in the light (visual motor neurons) or no activation in the dark (visual-dominant neurons). Of these visually responsive neurons, more than half (n = 66) responded during the object fixation task (object-type). Among these, 55 were tested for their shape selectivity during the object-fixation task, and many (n = 25) were highly selective, preferring one particular shape of the six different shapes presented (ring, cube, cylinder, cone, sphere, and square plate). For 28 moderately selective object-type neurons, we performed multidimensional scaling (MDS) to examine how the neurons encode the similarity of objects. The results suggest that some moderately selective neurons responded preferentially to common geometric features shared by similar objects (flat, round, elongated, etc.). Moderately selective nonobject-type visually responsive neurons, which did not respond during object fixation, were found by MDS to be more closely related to the handgrip than to the object shape. We found a similar selectivity for handgrip in motor-dominant neurons that did not show any visual response. With regard to the size of the objects, 16 of 26 object-type neurons tested were selective for both size and shape, whereas 9 object-type neurons were selective for shape but not for size. Seven of 12 nonobject-type and all (8/8) of the motor dominant neurons examined were selective for size, and almost all of them were also selective for objects. Many hand-manipulation-related neurons that preferred the plate and/or ring were selective for the orientation of the objects (17/20). These results suggest that the visual responses of object-type neurons represent the shape, size, and/or orientation of 3D objects, whereas those of the nonobject type neurons probably represent the shape of the handgrip, grip size, or hand orientation. The activity of motor-dominant neurons was also, in part, likely to represent these parameters of hand movement. This suggests that the dorsal visual pathway is concerned with the aspect of form, orientation, and/or size perception that is relevant for the visual control of movements. PMID- 10805660 TI - Position reconstruction from an ensemble of hippocampal place cells: contribution of theta phase coding. AB - Previous analysis of the firing of individual rat hippocampal place cells has shown that their firing rate increases when they enter a place field and that their phase of firing relative to the ongoing theta oscillation (7-12 Hz) varies systematically as the rat traverses the place field, a phenomenon termed the theta phase precession. To study the relative contribution of phased-coded and rate-coded information, we reconstructed the animal's position on a linear track using spikes recorded simultaneously from 38 hippocampal neurons. Two previous studies of this kind found no evidence that phase information substantially improves reconstruction accuracy. We have found that reconstruction is improved provided epochs with large, systematic errors are first excluded. With this condition, use of both phase and rate information improves the reconstruction accuracy by >43% as compared with the use of rate information alone. Furthermore, it becomes possible to predict the rat's position on a 204-cm track with very high accuracy (error of <3 cm). The best reconstructions were obtained with more than three phase divisions per theta cycle. These results strengthen the hypothesis that information in rat hippocampal place cells is encoded by the phase of theta at which cells fire. PMID- 10805661 TI - Dynorphin A elicits an increase in intracellular calcium in cultured neurons via a non-opioid, non-NMDA mechanism. AB - The opioid peptide dynorphin A is known to elicit a number of pathological effects that may result from neuronal excitotoxicity. An up-regulation of this peptide has also been causally related to the dysesthesia associated with inflammation and nerve injury. These effects of dynorphin A are not mediated through opioid receptor activation but can be effectively blocked by pretreatment with N-methyl-D-aspartate (NMDA) receptor antagonists, thus implicating the excitatory amino acid system as a mediator of the actions of dynorphin A and/or its fragments. A direct interaction between dynorphin A and the NMDA receptors has been well established; however the physiological relevance of this interaction remains equivocal. This study examined whether dynorphin A elicits a neuronal excitatory effect that may underlie its activation of the NMDA receptors. Calcium imaging of individual cultured cortical neurons showed that the nonopioid peptide dynorphin A(2-17) induced a time- and dose-dependent increase in intracellular calcium. This excitatory effect of dynorphin A(2-17) was insensitive to (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]-cyclohepten-5,10 imine (MK-801) pretreatment in NMDA-responsive cells. Thus dynorphin A stimulates neuronal cells via a nonopioid, non-NMDA mechanism. This excitatory action of dynorphin A could modulate NMDA receptor activity in vivo by enhancing excitatory neurotransmitter release or by potentiating NMDA receptor function in a calcium dependent manner. Further characterization of this novel site of action of dynorphin A may provide new insight into the underlying mechanisms of dynorphin excitotoxicity and its pathological role in neuropathy. PMID- 10805662 TI - Histamine suppresses non-NMDA excitatory synaptic currents in rat supraoptic nucleus neurons. AB - Whole cell patch-clamp recordings were obtained from supraoptic neurons to investigate the effects of histamine on excitatory postsynaptic currents evoked by electrical stimulation of areas around the posterior supraoptic nucleus. When cells were voltage-clamped at -70 mV, evoked excitatory postsynaptic currents had amplitudes of 88.4 +/- 9.6 pA and durations of 41.1 +/- 3.0 ms (mean +/- SE; n = 43). With twin stimulus pulses (20 Hz) used, paired-pulse facilitation ratios were 1.93 +/- 0.12. Bath application of 6-cyano-7-nitroquinoxalene-2,3-dione (CNQX) abolished synaptic currents. Histamine at concentrations approximately 0.1 10 microM reversibly suppressed excitatory postsynaptic currents in all supraoptic neurons tested. Within 2 min after application of (10 microM) histamine, current amplitudes and durations decreased by 61. 5 and 31.0%, respectively, with little change in the paired-pulse facilitation ratio. Dimaprit or imetit (H(2) or H(3) receptor agonists) did not reduce synaptic currents, whereas pyrilamine (H(1) receptor antagonist) blocked histamine-induced suppression of synaptic currents. When patch electrodes containing guanosine 5'-O (2-thiodiphosphate) (GDP-beta-S) were used to record cells, histamine still suppressed current amplitudes by 49.1% and durations by 41.9%. Similarly, intracellular diffusion of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) and H(7) did not abolish histamine-induced suppression of synaptic currents, either. Bath perifusion of 8-bromo-quanosine 3',5'-cyclic monophosphate reduced current amplitudes by 32.3% and durations by 27.9%. After bath perfusion of slices with N(omega)-nitro-L-arginine methyl ester (L-NAME), histamine injection decreased current amplitudes only by 31.9%, much less than the inhibition rate in control (P < 0.01). In addition, histamine induced little change in current durations and paired-pulse facilitation ratios, representing a partial blockade of histamine effects on synaptic currents by L-NAME. In supraoptic neurons recorded using electrodes containing BAPTA and perifused with L-NAME, the effects of histamine on synaptic currents were completely abolished. Norepinephrine injection reversibly decreased current amplitudes by 39.1% and duration by 64.5%, with a drop in the paired-pulse facilitation ratio of 47.9%. Bath perifusion of L-NAME, as well as intracellular diffusion of GDP-beta-S, 1-(5 isoquinolinylsulfonyl)-2-methyl-piperazine, or BAPTA, failed to block norepinephrine-induced suppression of evoked synaptic currents. The present results suggest that histamine suppresses non-N-methyl-D-aspartate synaptic currents in supraoptic neurons through activation of H(1) receptors. It is possible that histamine first acts at supraoptic cells (perhaps both neuronal and nonneuronal) and induces the production of nitric oxide, which then diffuses to nearby neurons and modulates synaptic transmission by a postsynaptic mechanism. PMID- 10805663 TI - Anatomy, physiology, and synaptic responses of rat layer V auditory cortical cells and effects of intracellular GABA(A) blockade. AB - The varied extracortical targets of layer V make it an important site for cortical processing and output, which may be regulated by differences in the pyramidal neurons found there. Two populations of projection neurons, regular spiking (RS) and intrinsic bursting (IB), have been identified in layer V of some sensory cortices, and differences in their inhibitory inputs have been indirectly demonstrated. In this report, IB and RS cells were identified in rat auditory cortical slices, and differences in thalamocortical inhibition reaching RS and IB cells were demonstrated directly using intracellular GABA(A) blockers. Thalamocortical synaptic input to RS cells was always a combination of excitation and both GABA(A) and GABA(B) inhibition. Stimulation seldom triggered a suprathreshold response. IB cell synaptic responses were mostly excitatory, and stimulation usually triggered action potentials. This apparent difference was confirmed directly using intracellular chloride channel blockers. Before intracellular diffusion, synaptic responses were stable and similar to control conditions. Subsequently, GABA(A) was blocked, revealing a cell's total excitatory input. On GABA(A) blockade, RS cells responded to synaptic stimulation with large, suprathreshold excitatory events, indicating that excitation, while always present in these cells, is masked by GABA(A). In IB cells that had visible GABA(A) input, it often masked an excitatory postsynaptic potential (EPSP) that could lead to additional suprathreshold events. These findings indicate that IB cells receive less GABA(A)-mediated inhibitory input and are able to spike or burst in response to thalamocortical synaptic stimulation far more readily than RS cells. Such differences may have implications for the influence each cell type exerts on its postsynaptic targets. PMID- 10805664 TI - Discrimination of line orientation in humans and monkeys. AB - Orientation discrimination, the capacity to recognize an orientation difference between two lines presented at different times, probably involves cortical processes such as stimuli encoding, holding them in memory, comparing them, and then deciding. To correlate discrimination with neural activity in combined psychophysical and electrophysiological experiments, precise knowledge of the strategies followed in the completion of the behavioral task is necessary. To address this issue, we measured human and nonhuman primates' capacities to discriminate the orientation of lines in a fixed and in a continuous variable task. Subjects have to indicate whether a line (test) was oriented to one side or to the other of a previously presented line (reference). When the orientation of the reference line did not change across trials (fixed discrimination task), subjects can complete the task either by categorizing the test line, thus ignoring the reference, or by discriminating between them. This ambiguity was avoided when the reference stimulus was changed randomly from trial to trial (continuous discrimination task), forcing humans and monkeys to discriminate by paying continuous attention to the reference and test stimuli. Both humans and monkeys discriminated accurately with stimulus duration as short as 150 ms. Effective interstimulus intervals were of 2.5 s for monkeys but much longer (>6 s) in humans. These results indicated that the fixed and continuous discrimination tasks are different, and accordingly humans and monkeys do use different behavioral strategies to complete each task. Because both tasks might involve different neural processes, these findings have important implications for studying the neural mechanisms underlying visual discrimination. PMID- 10805665 TI - Electrophysiological properties of cholinergic and noncholinergic neurons in the ventral pallidal region of the nucleus basalis in rat brain slices. AB - The ventral pallidum is a major source of output for ventral corticobasal ganglia circuits that function in translating motivationally relevant stimuli into adaptive behavioral responses. In this study, whole cell patch-clamp recordings were made from ventral pallidal neurons in brain slices from 6- to 18-day-old rats. Intracellular filling with biocytin was used to correlate the electrophysiological and morphological properties of cholinergic and noncholinergic neurons identified by choline acetyltransferase immunohistochemistry. Most cholinergic neurons had a large whole cell conductance and exhibited marked fast (i.e., anomalous) inward rectification. These cells typically did not fire spontaneously, had a hyperpolarized resting membrane potential, and also exhibited a prominent spike afterhyperpolarization (AHP) and strong spike accommodation. Noncholinergic neurons had a smaller whole cell conductance, and the majority of these cells exhibited marked time-dependent inward rectification that was due to an h-current. This current activated slowly over several hundred milliseconds at potentials more negative than -80 mV. Noncholinergic neurons fired tonically in regular or intermittent patterns, and two-thirds of the cells fired spontaneously. Depolarizing current injection in current clamp did not cause spike accommodation but markedly increased the firing frequency and in some cells also altered the pattern of firing. Spontaneous tetrodotoxin-sensitive GABA(A)-mediated inhibitory postsynaptic currents (IPSCs) were frequently recorded in noncholinergic neurons. These results show that cholinergic pallidal neurons have similar properties to magnocellular cholinergic neurons in other parts of the forebrain, except that they exhibit strong spike accommodation. Noncholinergic ventral pallidal neurons have large h-currents that could have a physiological role in determining the rate or pattern of firing of these cells. PMID- 10805666 TI - Differences in control of limb dynamics during dominant and nondominant arm reaching. AB - This study compares the coordination patterns employed for the left and right arms during rapid targeted reaching movements. Six right-handed subjects reached to each of three targets, designed to elicit progressively greater amplitude interaction torques at the elbow joint. All targets required the same elbow excursion (20 degrees ), but different shoulder excursions (5, 10, and 15 degrees, respectively). Movements were restricted to the shoulder and elbow and supported on a horizontal plane by a frictionless air-jet system. Subjects received visual feedback only of the final hand position with respect to the start and target locations. For motivation, points were awarded based on final position accuracy for movements completed within an interval of 400-600 ms. For all subjects, the right and left hands showed a similar time course of improvement in final position accuracy over repeated trials. After task adaptation, final position accuracy was similar for both hands; however, the hand trajectories and joint coordination patterns during the movements were systematically different. Right hand paths showed medial to lateral curvatures that were consistent in magnitude for all target directions, whereas the left hand paths had lateral to medial curvatures that increased in magnitude across the three target directions. Inverse dynamic analysis revealed substantial differences in the coordination of muscle and intersegmental torques for the left and right arms. Although left elbow muscle torque contributed largely to elbow acceleration, right arm coordination was characterized by a proximal control strategy, in which movement of both joints was primarily driven by the effects of shoulder muscles. In addition, right hand path direction changes were independent of elbow interaction torque impulse, indicating skillful coordination of muscle actions with intersegmental dynamics. In contrast, left hand path direction changes varied directly with elbow interaction torque impulse. These findings strongly suggest that distinct neural control mechanisms are employed for dominant and non dominant arm movements. However, whether interlimb differences in neural strategies are a consequence of asymmetric use of the two arms, or vice versa, is not yet understood. The implications for neural organization of voluntary movement control are discussed. PMID- 10805667 TI - Citrate enhances olfactory receptor responses and triggers oscillatory receptor activity in the channel catfish. AB - Citrate, a normal constituent of cellular metabolism, in a binary mixture with an amino acid enhanced asynchronous olfactory receptor responses in the channel catfish, Ictalurus punctatus. In addition, high concentrations of either citrate (> or =3 mM) alone or an amino acid (> or =0.1 mM) in a binary mixture with citrate (> or =1 mM) triggered synchronized voltage oscillations of olfactory receptor neurons (ORNs) known as "peripheral waves" (PWs). Binary mixtures containing lower concentrations of an amino acid also triggered PW activity if the concentration of citrate in the mixture was increased. Both the enhancement of asynchronous activity and the generation of PW activity were the result of citrate chelating calcium, which lowers the surface potential of ORNs making them hyperexcitable. These effects of citrate are replicated by EGTA. Inactivation of the chelating ability of citrate and EGTA with 1 mM calcium chloride, barium chloride, or strontium chloride abolished both the enhancement of asynchronous olfactory responses and PW activity, while not affecting olfactory receptor responses to the amino acids alone. PMID- 10805668 TI - Inhibition and disinhibition of pyramidal neurons by activation of nicotinic receptors on hippocampal interneurons. AB - Nicotinic acetylcholine receptors (nAChRs) are expressed in the hippocampus, and their functional roles are beginning to be delineated. The effect of nAChR activation on the activity of both interneurons and pyramidal neurons in the CA1 region was studied in rat hippocampal slices. In CA1 stratum radiatum with muscarinic receptors inhibited, local pressure application of acetylcholine (ACh) elicited a nicotinic current in 82% of the neurons. The majority of the ACh induced currents were sensitive to methyllycaconitine, which is a specific inhibitor of alpha7-containing nAChRs. Methyllycaconitine-insensitive nicotinic currents also were present as detected by a nonspecific nAChR inhibitor. The ACh sensitive neurons in the s. radiatum were identified as GABAergic interneurons by their electrophysiological properties. Pressure application of ACh induced firing of action potentials in approximately 70% of the interneurons. The ACh-induced excitation of interneurons could induce either inhibition or disinhibition of pyramidal neurons. The inhibition was recorded from the pyramidal neuron as a burst of GABAergic synaptic activity. That synaptic activity was sensitive to bicuculline, indicating that GABA(A) receptors mediated the ACh-induced synaptic currents. The disinhibition was recorded from the pyramidal neuron as a reduction of spontaneous GABAergic synaptic activity when ACh was delivered onto an interneuron. Both the inhibition and disinhibition were sensitive to either methyllycaconitine or mecamylamine, indicating that activation of nicotinic receptors on interneurons was necessary for the effects. These results show that nAChRs are capable of regulating hippocampal circuits by exciting interneurons and, subsequently, inhibiting or disinhibiting pyramidal neurons. PMID- 10805669 TI - Cloned delta-opioid receptors in GH(3) cells inhibit spontaneous Ca(2+) oscillations and prolactin release through K(IR) channel activation. AB - Opioid receptors can couple to K(+) and Ca(2+) channels, adenylyl cyclase, and phosphatidyl inositol turnover. Any of these actions may be important in the regulation of neurotransmitter and hormone release from excitable cells. GH(3) cells exhibit spontaneous oscillations of intracellular Ca(2+) concentration ([Ca(2+)](i)) and prolactin release. Activation of cloned delta-opioid receptors stably expressed in GH(3) cells inhibits both spontaneous Ca(2+) signaling and basal prolactin release. The objective of this study was to examine a possible role for K(+) channels in these processes using the patch-clamp technique, fluorescence imaging, and a sensitive ELISA for prolactin. The selective delta receptor agonist [D-Pen(2), D-Pen(2)]enkephalin (DPDPE) inhibited [Ca(2+)](i) oscillations in GH(3) cells expressing both mu and delta receptors (GH(3)MORDOR cells) but had no effect on control GH(3) cells or cells expressing mu receptors alone (GH(3)MOR cells). The inhibition of [Ca(2+)](i) oscillations by DPDPE was unaffected by thapsigargin pretreatment, suggesting that this effect is independent of inositol 1,4,5-triphosphate-sensitive Ca(2+) stores. DPDPE caused a concentration-dependent inhibition of prolactin release from GH(3)MORDOR cells with an IC(50) of 4 nM. DPDPE increased inward K(+) current recorded from GH(3)MORDOR cells but had no significant effect on K(+) currents recorded from control GH(3) cells or GH(3)MOR cells. The mu receptor agonist morphine also had no effect on currents recorded from control cells but activated inward K(+) currents recorded from GH(3)MOR and GH(3)MORDOR cells. Somatostatin activated inward currents recorded from all three cell lines. The DPDPE-sensitive K(+) current was inwardly rectifying and was inhibited by Ba(2+) but not TEA. DPDPE had no effect on delayed rectifier-, Ca(2+)-, and voltage-activated or A-type K(+) currents, recorded from GH(3)MORDOR cells. Ba(2+) attenuated the inhibition of [Ca(2+)](i) and prolactin release by DPDPE, whereas TEA had no effect, consistent with an involvement of K(IR) channels in these actions of the opioid. PMID- 10805670 TI - Ongoing and stimulus-evoked activity of sympathetically correlated neurons in the intermediate zone and dorsal horn of acutely spinalized rats. AB - We have shown previously that in the acutely spinalized anesthetized rat the activities of many dorsal horn interneurons (DHN) at the T(10) level are correlated positively with both ongoing and stimulus-evoked renal sympathetic nerve activity (RSNA) and therefore may belong to networks generating RSNA after acute, cervical, spinal transection. In the present study, we recorded from both DHN and interneurons in the intermediate zone (IZN) of the T(10) spinal segment in acutely C(1)-transected, chloralose-anesthetized, artificially respired rats. The activities of a similar percentage of IZN and DHN were correlated positively with ongoing RSNA, but the peaks of spike-triggered averages of RSNA based on the activity of IZN were larger, relative to dummy averages, than spike-triggered averages of RSNA based on the activity of DHN. Sympathetically correlated DHN and IZN differed in their responses to noxious somatic stimuli. Most correlated DHN had relatively simple somatic fields; they were excited by noxious stimulation of the T(10) and nearby dermatomes and inhibited by stimulation of more distal dermatomes. As we have shown previously, the excitatory and inhibitory fields of these neurons were very similar to fields that, respectively, excited and inhibited RSNA. On the other hand, the somatic fields of 50% of sympathetically correlated IZN were significantly more complex, indicating a difference between either the inputs or the processing properties of IZN and DHN. Sympathetically correlated IZN and DHN also differed in their responses to colorectal distension (CRD), a noxious visceral stimulus. CRD increased RSNA in 11/15 rats and increased the activity of most sympathetically correlated T(10) IZN. On the other hand, CRD decreased the activity of a majority of sympathetically correlated T(10) DHN. These observations suggest that the same stimulus may differentially affect separate, putative, sympathoexcitatory pathways, exciting one and inhibiting the other. Thus the magnitude and even the polarity of responses to a given stimulus may be determined by the modality and location of the stimulus, the degree to which multiple pathways are affected by the stimulus, and the ongoing activity of presympathetic neurons, at multiple rostrocaudal levels, before stimulation. A multipathway system may explain the variability in autonomic responses to visceral and somatic stimuli exhibited in spinally injured patients. PMID- 10805671 TI - Sound-induced synchronization of neural activity between and within three auditory cortical areas. AB - Neural synchrony within and between auditory cortical fields is evaluated with respect to its potential role in feature binding and in the coding of tone and noise sound pressure level. Simultaneous recordings were made in 24 cats with either two electrodes in primary auditory cortex (AI) and one in anterior auditory field (AAF) or one electrode each in AI, AAF, and secondary auditory cortex. Cross-correlograms (CCHs) for 1-ms binwidth were calculated for tone pips, noise bursts, and silence (i.e., poststimulus) as a function of intensity level. Across stimuli and intensity levels the total percentage of significant stimulus onset CCHs was 62% and that of significant poststimulus CCHs was 58% of 1,868 pairs calculated for each condition. The cross-correlation coefficient to stimulus onsets was higher for single-electrode pairs than for dual-electrode pairs and higher for noise bursts compared with tone pips. The onset correlation for single-electrode pairs was only marginally larger than the poststimulus correlation. For pairs from electrodes across area boundaries, the onset correlations were a factor 3-4 higher than the poststimulus correlations. The within-AI dual-electrode peak correlation was higher than that across areas, especially for spontaneous conditions. Correlation strengths for between area pairs were independent of the difference in characteristic frequency (CF), thereby providing a mechanism of feature binding for broadband sounds. For noise burst stimulation, the onset correlation for between area pairs was independent of stimulus intensity regardless the difference in CF. In contrast, for tone-pip stimulation a significant dependence on intensity level of the peak correlation strength was found for pairs involving AI and/or AAF with CF difference less than one octave. Across all areas, driven rate, between-area peak correlation strength, or a combination of the two did not predict stimulus intensity. However, between-area peak correlation strength performs better than firing rate to decide if a stimulus is present or absent. PMID- 10805672 TI - Correlation between the activity of single auditory cortical neurons and sound localization behavior in the macaque monkey. AB - Lesion studies have indicated that the auditory cortex is crucial for the perception of acoustic space, yet it remains unclear how these neurons participate in this perception. To investigate this, we studied the responses of single neurons in the primary auditory cortex (AI) and the caudomedial field (CM) of two monkeys while they performed a sound-localization task. Regression analysis indicated that the responses of approximately 80% of neurons in both cortical areas were significantly correlated with the azimuth or elevation of the stimulus, or both, which we term "spatially sensitive." The proportion of spatially sensitive neurons was greater for stimulus azimuth compared with stimulus elevation, and elevation sensitivity was primarily restricted to neurons that were tested using stimuli that the monkeys also could localize in elevation. Most neurons responded best to contralateral speaker locations, but we also encountered neurons that responded best to ipsilateral locations and neurons that had their greatest responses restricted to a circumscribed region within the central 60 degrees of frontal space. Comparing the spatially sensitive neurons with those that were not spatially sensitive indicated that these two populations could not be distinguished based on either the firing rate, the rate/level functions, or on their topographic location within AI. Direct comparisons between the responses of individual neurons and the behaviorally measured sound localization ability indicated that proportionally more neurons in CM had spatial sensitivity that was consistent with the behavioral performance compared with AI neurons. Pooling the responses across neurons strengthened the relationship between the neuronal and psychophysical data and indicated that the responses pooled across relatively few CM neurons contain enough information to account for sound-localization ability. These data support the hypothesis that auditory space is processed in a serial manner from AI to CM in the primate cerebral cortex. PMID- 10805673 TI - Membrane properties of chick semicircular canal hair cells in situ during embryonic development. AB - The electrophysiological properties of developing vestibular hair cells have been investigated in a chick crista slice preparation, from embryonic day 10 (E10) to E21 (when hatching would occur). Patch-clamp whole-cell experiments showed that different types of ion channels are sequentially expressed during development. An inward Ca(2+) current and a slow outward rectifying K(+) current (I(K(V))) are acquired first, at or before E10, followed by a rapid transient K(+) current (I(K(A))) at E12, and by a small Ca-dependent K(+) current (I(KCa)) at E14. Hair cell maturation then proceeds with the expression of hyperpolarization-activated currents: a slow I(h) appears first, around E16, followed by the fast inward rectifier I(K1) around E19. From the time of its first appearance, I(K(A)) is preferentially expressed in peripheral (zone 1) hair cells, whereas inward rectifying currents are preferentially expressed in intermediate (zone 2) and central (zone 3) hair cells. Each conductance conferred distinctive properties on hair cell voltage response. Starting from E15, some hair cells, preferentially located at the intermediate region, showed the amphora shape typical of type I hair cells. From E17 (a time when the afferent calyx is completed) these cells expressed I(K, L), the signature current of mature type I hair cells. Close to hatching, hair cell complements and regional organization of ion currents appeared similar to those reported for the mature avian crista. By the progressive acquisition of different types of inward and outward rectifying currents, hair cell repolarization after both positive- and negative-current injections is greatly strengthened and speeded up. PMID- 10805674 TI - Visually evoked cyclovergence and extended listing's law. AB - Cyclovergence is a simultaneously occurring cyclorotation of the two eyes in opposite directions. Cyclovergence can be elicited visually by opposite cyclorotation of the two eyes' images. It also can occur in conjunction with horizontal vergence and vertical version in a stereotyped manner as described by the extended Listing's law (or L2). We manipulated L2-related and visually evoked cyclovergence independently, using stereoscopic images of three-dimensional (3D) scenes. During pursuit in the midsagittal plane, cyclovergence followed L2. The amount of L2-related cyclovergence during pursuit varied between subjects. Each pursuit trial was repeated three times. Two of the three trials had additional image rotation to visually evoke cyclovergence. We could separate the L2-related and visual components of cyclovergence by subtraction of the cyclovergence response in matched trials that differed only in the image rotation that was applied during pursuit. This indicates that visual and L2-related contributions to cyclovergence add linearly, suggesting the presence of two independent systems. Visually evoked cyclovergence gains were characteristic for a given subject, little affected by visual stimulus parameters, and usually low (0.1-0.5) when a static target was fixated. Gain and phase lag of the visually evoked cyclovergence during vertical pursuit was comparable with that during fixation of a static target. The binocular orientations are in better agreement to orientations predicted by L2 then would be predicted by nulling of the cyclodisparities. On the basis of our results, we suggest that visually driven and L2-related cyclovergence are independent of each other and superimpose linearly. PMID- 10805675 TI - Possible involvement of undissociated acid molecules in the acid response of the chorda tympani nerve of the rat. AB - To test whether undissociated acid is capable of exciting the chorda tympani nerves in rats, we have used buffered acid solutions as taste stimuli. These solutions were prepared by adding alkali to weak acids, such as acetic acid, so that the proportion of undissociated and dissociated acids was varied whereas keeping the total acid concentration constant. When acetic acid solutions, adjusted to wide ranges of pH by NaOH, were applied to the tongue, the response magnitude of the chorda tympani nerves was not varied systematically with pH changes. However, if the sodium effect was eliminated by amiloride or replacement of cation by potassium or Tris[hydroxymethyl]aminomethane; NH(2)C(CH(2)OH)(3) (Tris-base), the chorda tympani response was reduced systematically as pH increased. Similar results were obtained with citric acid and ascorbic acid. This pH-dependent change in taste nerve response to acid cannot be solely attributed to the proton gradient because the response magnitude induced by hydrogen itself, which was estimated from responses to strong acids, was much smaller than that by equi-pH acetic acid ( approximately 85%). Thus we cannot explain the pH-dependent responses of the chorda tympani nerves to weak acids unless effects of undissociated acid molecules are postulated. It is therefore concluded that undissociated acids in weak acid solutions can be a stimulant to taste receptor cells. PMID- 10805676 TI - Temporary inactivation in the primate motor thalamus during visually triggered and internally generated limb movements. AB - To better understand the contribution of cerebellar- and basal ganglia-receiving areas of the thalamus [ventral posterolateral nucleus, pars oralis (VPLo), area X, ventral lateral nucleus, pars oralis (VLo), or ventral anterior nucleus, pars parvicellularis (VApc)] to movements based on external versus internal cues, we temporarily inactivated these individual nuclei in two monkeys trained to make visually triggered (VT) and internally generated (IG) limb movements. Infusions of lignocaine centered within VPLo caused hemiplegia during which movements of the contralateral arm rarely were performed in either task for a short period of time ( approximately 5-30 min). When VT responses were produced, they had prolonged reaction times and movement times and a higher incidence of trajectory abnormalities compared with responses produced during the preinfusion baseline period. In contrast, those IG responses that were produced remained relatively normal. Infusions centered within area X never caused hemiplegia. The only deficits observed were an increase in reaction time and movement amplitude variability and a higher incidence of trajectory abnormalities during VT trials. Every other aspect of both the VT and IG movements remained unchanged. Infusions centered within VLo reduced the number of movements attempted during each block of trials. This did not appear to be due to hemiplegia, however, as voluntary movements easily could be elicited outside of the trained tasks. The other main deficit resulting from inactivation of VLo was an increased reaction time in the VT task. Finally, infusions centered within VApc caused IG movements to become slower and smaller in amplitude, whereas VT movements remained unchanged. Control infusions with saline did not cause any consistent deficits. This pattern of results implies that VPLo and VLo play a role in the production of movements in general regardless of the context under which they are performed. They also suggest that VPLo contributes more specifically to the execution of movements that are visually triggered and guided, whereas area X contributes specifically to the initiation of such movements. In contrast, VApc appears to play a role in the execution of movements based on internal cues. These results are consistent with the hypothesis that specific subcircuits within the cerebello- and basal ganglio-thalamo-cortical systems preferentially contribute to movements based on external versus internal cues. PMID- 10805677 TI - High responsiveness and direction sensitivity of neurons in the rat thalamic reticular nucleus to vibrissa deflections. AB - The thalamic reticular nucleus (Rt) is strategically positioned to integrate descending and ascending signals in the control of sensorimotor and other thalamocortical activity. Its prominent role in the generation of sleep spindles notwithstanding, relatively little is known of Rt function in regulating interactions with the sensory environment. We recorded and compared the responses of individual Rt and thalamocortical neurons in the ventroposterior medial (VPm) nucleus of the rat to controlled deflections of mystacial vibrissae. Transient Rt responses to the onset (ON) and offset (OFF) of vibrissa deflection are larger and longer in duration than those of VPm and of all other populations studied in the whisker/barrel pathway. Magnitudes of ON and OFF responses in Rt were negatively correlated with immediately preceding activities, suggesting a contribution of low-threshold T-type Ca(2+) channels. Rt neurons also respond with high tonic firing rates during sustained vibrissa deflections. By comparison, VPm neurons are less likely to respond tonically and are more likely to exhibit tonic suppression. Rt and VPm populations are similar to each other, however, in that they retain properties of directional sensitivity established in primary afferent neurons. In both populations neurons are selective for deflection angle and exhibit directional consistency, responding best to a particular direction of movement regardless of the starting position of the vibrissal hair. These findings suggest a role for Rt in the processing of detailed sensory information. Temporally, Rt may function to limit the duration of stimulus-evoked VPm responses and to focus them on rapid vibrissa perturbations. Moreover, by regulating the baseline activity of VPm neurons, Rt may indirectly enhance the response selectivity of layer IV barrel neurons to synchronous VPm firing. PMID- 10805678 TI - The influence of single VB thalamocortical impulses on barrel columns of rabbit somatosensory cortex. AB - Extracellular recordings were obtained from single neurons in ventrobasal (VB) thalamus of awake rabbits while field potentials were recorded at various depths within topographically aligned and nonaligned barrel columns of somatosensory cortex (S1). Spike-triggered averages of cortical field potentials were obtained following action potentials in thalamic neurons. Action potentials in a VB neuron elicited a cortical response within layer 4 with three distinct components. 1) A biphasic, initially positive response (latency <1 ms) was interpreted to reflect activation of the VB axon terminals (the AxTP). This response was not affected by infusion of an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor antagonist within the barrel. In contrast, later components of the response were completely eliminated and were interpreted to reflect focal synaptic potentials. 2) A negative potential [focal synaptic negativity (FSN)] occurred at a mean latency of 1.65 ms and lasted approximately 4 ms. This response had a rapid rise time ( approximately 0.7 ms) and was interpreted to reflect monosynaptic excitation. 3) The third component was a positive potential (the FSP), with a slow rise time and a half-amplitude duration of approximately 30 ms. The FSP showed a weak reversal in superficial cortical layers and was interpreted to reflect di/polysynaptic inhibition. The amplitudes of the AxTP, the FSN, and the FSP reached a peak near layer 4 and were highly attenuated in both superficial and deep cortical layers. All components were attenuated or absent when the cortical electrode was missaligned from the thalamic electrode by a single cortical barrel. Deconvolution procedures revealed that the autocorrelogram of the presynaptic VB neuron had very little influence on either the amplitude or duration of the AxTP or the FSN, and only a minor influence (mean, 11%) on the amplitude of the FSP. We conclude that individual VB thalamic impulses entering a cortical barrel engage both monosynaptic excitatory and di/polysynaptic inhibitory mechanisms. Putative inhibitory interneurons of an S1 barrel receive a highly divergent/convergent monosynaptic input from the topographically aligned VB barreloid, and this results in sharp synchrony among these interneurons. We suggest that single-fiber access to disynaptic inhibition is facilitated by this sharp synchrony, and that the FSP reflects a consequent synchronous wave of feed-forward inhibition within the S1 barrel. PMID- 10805679 TI - Could different directions of infant stepping be controlled by the same locomotor central pattern generator? AB - This study examined the idea of whether the same central pattern generator (CPG) for locomotion can control different directions of walking in humans. Fifty-two infants, aged 2-11 mo, were tested. Infants were supported to walk on a treadmill at a variety of speeds. If forward stepping was elicited, stepping in the other directions (primarily sideways and backward) was attempted. The orientation of the infant on the treadmill belt determined the direction of stepping. In some infants, we also attempted to obtain a smooth transition from one direction to another by gradually changing the orientation of the infant during a stepping sequence. Limb segment motion and surface electromyography from the muscles of the lower limb were recorded. Most infants who showed sustained forward walking also could walk in all other directions. Thirty-three of 34 infants tested could step sideways. The success of eliciting backward stepping was 69%. Most of the infants who did not meet our backward stepping criteria did, however, make stepping movements. The different directions of stepping had similar responses to changes in treadmill speed. The relationship between stance and swing phase durations and cycle duration were the same regardless of the direction of stepping across a range of speeds. Some differences were noted in the muscle activation patterns during different directions of walking. For example, the hamstrings were much more active during the swing phase of backward walking compared with forward walking. The quadriceps was more active in the trailing leg during sideways walking. In some infants, we were able to elicit stepping along a continuum of directions. We found no discrete differences in either the electromyographic patterns or the temporal parameters of stepping as the direction of stepping was gradually changed. The results support the idea that the same locomotor CPG controls different directions of stepping in human infants. The fact that most infants were able to step in all directions, the similarity in the response to speed changes, and the absence of any discrete changes as the direction of stepping was changed gradually are all consistent with this hypothesis. PMID- 10805680 TI - Spike coding during locomotor network activity in ventrally located neurons in the isolated spinal cord from neonatal rat. AB - To characterize spike coding in spinal neurons during rhythmic locomotor activity, we recorded from individual cells in the lumbar spinal cord of neonatal rats by using the on-cell patch-clamp technique. Locomotor activity was induced by N-methyl-D aspartate (NMDA) and 5-hydroxytryptamine (5-HT) and monitored by ventral root recording. We made an estimator based on the assumption that the number of spikes arriving during two halves of the locomotor cycle could be a code used by the neuronal network to distinguish between the halves. This estimator, termed the spike contrast, was calculated as the difference between the number of spikes in the most and least active half of an average cycle. The root activity defined the individual cycles and the positions of the spikes were calculated relative to these cycles. By comparing the average spike contrast to the spike contrast in noncyclic, randomized spike trains we found that approximately one half the cells (19 of 42) contained a significant spike contrast, averaging 1.25 +/- 0.23 (SE) spikes/cycle. The distribution of spike contrasts in the total population of cells was exponential, showing that weak modulation was more typical than strong modulation. To investigate if this low spike contrast was misleading because a higher spike contrast averaged out by occurring at different positions in the individual cycles we compared the spike contrast obtained from the average cycle to its maximal value in the individual cycles. The value was larger (3.13 +/- 0.25 spikes) than the spike contrast in the average cycle but not larger than the spike contrast in the individual cycles of a random, noncyclic spike trains (3.21 +/- 0.21 spikes). This result suggested that the important distinction between cyclic and noncyclic cells was only the repeated cycle position of the spike contrast and not its magnitude. Low spike frequencies (5.2 +/- 0.82 spikes/cycle, that were on average 3.5 s long) and a minimal spike interval of 100-200 ms limited the spike contrast. The standard deviation (SD) of the spike contrast in the individual neurons was similar to the average spike contrasts and was probably stochastic because the SDs of the simulated, noncyclic spike trains were also similar. In conclusion we find a highly distributed and variable locomotor related cyclic signal that is represented in the individual neurons by very few spikes and that becomes significant only because the spike contrast is repeated at a preferred phase of the locomotor cycle. PMID- 10805681 TI - Synaptically released glutamate does not overwhelm transporters on hippocampal astrocytes during high-frequency stimulation. AB - In addition to maintaining the extracellular glutamate concentration at low ambient levels, high-affinity glutamate transporters play a direct role in synaptic transmission by speeding the clearance of glutamate from the synaptic cleft and limiting the extent to which transmitter spills over between synapses. Transporters are expressed in both neurons and glia, but glial transporters are likely to play the major role in removing synaptically released glutamate from the extracellular space. The role of transporters in synaptic transmission has been studied directly by measuring synaptically activated, transporter-mediated currents (STCs) in neurons and astrocytes. Here we record from astrocytes in the CA1 region of hippocampal slices and elicit STCs with high-frequency (100 Hz) stimulus trains of varying length to determine whether transporters are overwhelmed by stimuli that induce long-term potentiation. We show that, at near physiological temperatures (34 degrees C), high-frequency stimulation (HFS) does not affect the rate at which transporters clear glutamate from the extrasynaptic space. Thus, although spillover between synapses during "normal" stimulation may compromise the absolute synapse specificity of fast excitatory synaptic transmission, spillover is not exacerbated during HFS. Transporter capacity is diminished somewhat at room temperature (24 degrees C), although transmitter released during brief, "theta burst" stimulation is still cleared as quickly as following a single stimulus, even when transport capacity is partially diminished by pharmacological means. PMID- 10805682 TI - Group I mGluR activation turns on a voltage-gated inward current in hippocampal pyramidal cells. AB - A unique property of the group I metabotropic glutamate receptor (mGluR)-induced depolarization in hippocampal cells is that the amplitude of the depolarization is larger when the response is elicited at more depolarized membrane potentials. Our understanding of the conductance mechanism underlying this voltage-dependent response is incomplete. Through the use of current-clamp and single-electrode voltage-clamp recordings in guinea pig hippocampal slices, we examined the group I mGluR-induced depolarization in CA3 pyramidal cells. The group I mGluR agonists (S)-3-hydroxyphenylglycine and (S)-3,5-dihydroxyphenylglycine turned on a voltage gated inward current (I(mGluR(V))), which was pharmacologically distinct from the voltage-gated sodium and calcium currents intrinsic to the cells. I(mGluR(V)) was a slowly activating, noninactivating current with a threshold at about -75 mV. In addition to the activation of I(mGluR(V)), group I mGluR stimulation also produced a voltage-independent decrease in the K(+) conductance. Our results suggest that the depolarization induced by group I mGluR activation is generated by two ionic mechanisms-a heretofore unrecognized voltage-gated inward current (I(mGluR(V))) that is turned on by depolarization and a voltage-insensitive inward current that results from a turn-off of the K(+) conductance. The low threshold and noninactivating properties of I(mGluR(V)) allow the current to play a significant role in setting the resting potential and firing pattern of CA3 pyramidal cells. PMID- 10805683 TI - Pre-Botzinger complex functions as a central hypoxia chemosensor for respiration in vivo. AB - Recently, we identified a region located in the pre-Botzinger complex (pre-BotC; the proposed locus of respiratory rhythm generation) in which activation of ionotropic excitatory amino acid receptors using DL-homocysteic acid (DLH) elicits a variety of excitatory responses in the phrenic neurogram, ranging from tonic firing to a rapid series of high-amplitude, rapid rate of rise, short duration inspiratory bursts that are indistinguishable from gasps produced by severe systemic hypoxia. Therefore we hypothesized that this unique region is chemosensitive to hypoxia. To test this hypothesis, we examined the response to unilateral microinjection of sodium cyanide (NaCN) into the pre-BotC in chloralose- or chloralose/urethan-anesthetized vagotomized, paralyzed, mechanically ventilated cats. In all experiments, sites in the pre-BotC were functionally identified using DLH (10 mM, 21 nl) as we have previously described. All sites were histologically confirmed to be in the pre-BotC after completion of the experiment. Unilateral microinjection of NaCN (1 mM, 21 nl) into the pre-BotC produced excitation of phrenic nerve discharge in 49 of the 81 sites examined. This augmentation of inspiratory output exhibited one of the following changes in cycle timing and/or pattern: 1) a series of high-amplitude, short-duration bursts in the phrenic neurogram (a discharge similar to a gasp), 2) a tonic excitation of phrenic neurogram output, 3) augmented bursts in the phrenic neurogram (i.e., eupneic breath ending with a gasplike burst), or 4) an increase in frequency of phrenic bursts accompanied by small increases or decreases in the amplitude of integrated phrenic nerve discharge. Our findings identify a locus in the brain stem in which focal hypoxia augments respiratory output. We propose that the respiratory rhythm generator in the pre-BotC has intrinsic hypoxic chemosensitivity that may play a role in hypoxia-induced gasping. PMID- 10805684 TI - Locations of spinothalamic tract axons in cervical and thoracic spinal cord white matter in monkeys. AB - The spinothalamic tract (STT) is the primary pathway carrying nociceptive information from the spinal cord to the brain in humans. The aim of this study was to understand better the organization of STT axons within the spinal cord white matter of monkeys. The location of STT axons was determined using method of antidromic activation. Twenty-six lumbar STT cells were isolated. Nineteen were classified as wide dynamic range neurons and seven as high-threshold cells. Fifteen STT neurons were recorded in the deep dorsal horn (DDH) and 11 in superficial dorsal horn (SDH). The axons of 26 STT neurons were located at 73 low threshold points (<30 microA) within the lateral funiculus from T(9) to C(6). STT neurons in the SDH were activated from 33 low-threshold points, neurons in the DDH from 40 low-threshold points. In lower thoracic segments, SDH neurons were antidromically activated from low-threshold points at the dorsal-ventral level of the denticulate ligament. Neurons in the DDH were activated from points located slightly ventral, within the ventral lateral funiculus. At higher segmental levels, axons from SDH neurons continued in a position dorsal to those of neurons in the DDH. However, axons from neurons in both areas of the gray matter were activated from points located in more ventral positions within the lateral funiculus. Unlike the suggestions in several previous reports, the present findings indicate that STT axons originating in the lumbar cord shift into increasingly ventral positions as they ascend the length of the spinal cord. PMID- 10805685 TI - On the origin of the soleus H-reflex modulation pattern during human walking and its task-dependent differences. AB - Recently, Brooke and colleagues have suggested "that the strong inhibition arising from passive movement about the knee and hip joints, lays down the base for the soleus H-reflex gain modulation seen during human gait." In particular stretch-evoked afferent activity from the quadriceps muscle was emphasized as the most important source of movement-induced inhibition of the H-reflex. To test this hypothesis we examined the kinematics and electromyographic (EMG) activity of the leg during human walking and correlated these with the modulation pattern of the soleus H-reflex. To further test the possible contribution of stretch evoked quadriceps afferent activity to the soleus H-reflex modulation pattern during walking different walking gaits were studied. In one condition subjects were asked to walk with their knee locked in full extension by a rigid knee brace. In a second condition subjects were asked to walk backwards. During normal walking, the soleus H-reflex modulation pattern is strongly correlated with the EMG events of the soleus and tibialis anterior (TA), but not with hip, knee, or ankle angular displacement or velocity. When subjects walked with the knee locked in full extension, the amplitude of the H-reflex, its modulation pattern, and the task-dependent changes of its amplitude were the same as during normal walking. During backward walking, the H-reflex increases in late swing before activity of the soleus has begun and while the knee is flexing, an observation that highlights central control of the H-reflex amplitude. The effects of imposed flexion of the knee in passive subjects were also reexamined. The knee flexion imposed by the experimenter followed the same trajectory as that which occurred during the swing phase of the subject's step cycle. It was found that imposed knee flexions elicited a burst of TA EMG activity with an average latency of 81.6 ms (SD = 21 ms) in six out of eight subjects. Inhibition of the H-reflex, when it occurred, was associated with the occurrence of this burst. When subjects voluntarily flexed their right knee from an initial quiet standing posture, the inhibition of the soleus H-reflex began before flexion of the knee or that of any other leg segment. Once again the onset of inhibition was closely associated with the onset of activity in the TA. In the discussion section the present observations are examined in light of the predictions made by the movement induced inhibition hypothesis of Brooke et al. It will be concluded that none of the predictions of this hypothesis were corroborated by present tests done during human walking. In consequence, we suggest that the modulation pattern of the H reflex observed during normal human walking is centrally determined, as are the task-dependent differences of its amplitude (e.g., standing versus the stance phase of human walking). PMID- 10805686 TI - Input of anterior and posterior semicircular canal interneurons encoding head velocity to the dorsal Y group of the vestibular nuclei. AB - Neurons in the Y group of the vestibular nuclei are activated disynaptically from the ipsilateral VIIIth nerve and polysynaptically from the contralateral nerve. The ipsilateral anterior and posterior semicircular canals project to the Y group via interneurons in the vestibular nuclei. Candidate interneurons located in the rostrolateral corner of the superior (SVN) and in the caudal medial (MVN) vestibular nuclei were retrogradely labeled by the iontophoretic injection of biocytin into the Y group. The physiology of these interneurons named Y-group projecting neurons (YPNs) was studied in the SVN. SVN-YPNs were activated antidromically by electric pulse stimulation in the Y group. The properties of SVN-YPNs are distinct from those of SVN flocculus projecting neurons (FPNs). Namely, YPNs have a lower resting rate than FPNs, have more irregular interspike intervals, show a different phase and gain during the vestibuloocular reflex, and are located differentially within the SVN. After the injection of biocytin into the Y group, the locations of Purkinje cells that project to the Y group were confined to the vertical zones of the flocculus and ventral paraflocculus. However, mossy fibers originating in the Y group terminate in both the vertical and horizontal zones of the flocculus and ventral paraflocculus as well as in the ipsilateral nodulus. PMID- 10805687 TI - Ischemia but not anoxia evokes vesicular and Ca(2+)-independent glutamate release in the dorsal vagal complex in vitro. AB - Whole cell recordings of fura-2 dialyzed vagal neurons of brain stem slices were used to monitor interstitial glutamate accumulation within the dorsal vagal complex. Anoxia produced a sustained outward current (60 pA) and a moderate [Ca(2+)](i) rise (40 nM). These responses were neither mimicked by [1S,3R]-1 aminocyclo-pentane-1, 3-dicarboxylic acid nor affected by Ca(2+)-free solution, 6 cyano-7-nitroquino-xaline-2,3-dione (CNQX), 2-amino-5-phosphonovalerate (APV), or tetrodotoxin. Anoxia or cyanide in glucose-free saline (in vitro ischemia) as well as ouabain or iodoacetate elicited an initial anoxia-like [Ca(2+)](i) increase that turned after several minutes into a prominent Ca(2+) transient (0.9 microM) and inward current (-1.8 nA). APV plus CNQX (plus methoxyverapamil) inhibited this inward current as well as accompanying spontaneous synaptic activity, and reduced the secondary [Ca(2+)](i) rise to values similar to those during anoxia. Each of the latter drugs delayed onset of both ischemic current and prominent [Ca(2+)](i) rise by several minutes and attenuated their magnitudes by up to 40%. Ca(2+)-free solution induced a twofold delay of the ischemic inward current and suppressed the prominent Ca(2+) increase but not the initial moderate [Ca(2+)](i) rise. Cyclopiazonic acid or arachidonic acid in Ca(2+)-free saline delayed further the ischemic current, whereas neither inhibitors of glutamate uptake (dihydrokainate, D,L-threo-beta-hydroxyaspartate, L-transpyrrolidone-2,4 dicarboxylate) nor the Cl(-) channel blocker 5-nitro-2-(3-phenylpropyl-amino) benzoic acid had any effect. In summary, the response to metabolic arrest is due to activation of ionotropic glutamate receptors causing Ca(2+) entry via N-methyl D-aspartate receptors and voltage-activated Ca(2+) channels. An early Ca(2+) dependent exocytotic phase of ischemic glutamate release is followed by nonvesicular release, not mediated by reversed glutamate uptake or Cl(-) channels. The results also show that glycolysis prevents glutamate release during anoxia. PMID- 10805688 TI - Selective depolarization of interneurons in the early posttraumatic dentate gyrus: involvement of the Na(+)/K(+)-ATPase. AB - Interneurons innervating dentate granule cells are potent regulators of the entorhino-hippocampal interplay. Traumatic brain injury, a leading cause of death and disability among young adults, is frequently associated with rapid neuropathological changes, seizures, and short-term memory deficits both in humans and experimental animals, indicating significant posttraumatic perturbations of hippocampal circuits. To determine the pathophysiological alterations that affect the posttraumatic functions of dentate neuronal networks within the important early (hours to days) posttraumatic period, whole cell patch clamp recordings were performed from granule cells and interneurons situated in the granule cell layer of the dentate gyrus of head-injured and age-matched, sham operated control rats. The data show that a single pressure wave-transient delivered to the neocortex of rats (mimicking moderate concussive head trauma) resulted in a characteristic ( approximately 10 mV), transient (<4 days), selective depolarizing shift in the resting membrane potential of dentate interneurons, but not in neighboring granule cells. The depolarization was not associated with significant changes in action potential characteristics or input resistance, and persisted in the presence of antagonists of ionotropic and metabotropic glutamate, and GABA(A) and muscarinic receptors, as well as blockers of voltage-dependent sodium channels and of the h-current. The differential action of the cardiac glycosides oubain and stophanthidin on interneurons from control versus head-injured rats indicated that the depolarization of interneurons was related to the trauma-induced decrease in the activity of the electrogenic Na(+)/K(+)-ATPase. In contrast, the Na(+)/K(+)-ATPase activity in granule cells did not change. Intracellular injection of Na(+), Ca(2+)-chelator and ATP, as well as ATP alone, abolished the difference between the resting membrane potentials of control and injured interneurons. The selective posttraumatic depolarization increased spontaneous firing in interneurons, enhanced the frequency and amplitude of spontaneous inhibitory postsynaptic currents (IPSCs) in granule cells, and augmented the efficacy of depolarizing inputs to discharge interneurons. These results demonstrate that mechanical neurotrauma delivered to a remote site has highly selective effects on different cell types even within the same cell layer, and that the electrogenic Na(+)-pump plays a role in setting the excitability of hippocampal interneuronal networks after injury. PMID- 10805689 TI - Reference frames for spinal proprioception: limb endpoint based or joint-level based? AB - Many sensorimotor neurons in the CNS encode global parameters of limb movement and posture rather than specific muscle or joint parameters. Our investigations of spinocerebellar activity have demonstrated that these second-order spinal neurons also may encode proprioceptive information in a limb-based rather than joint-based reference frame. However, our finding that each foot position was determined by a unique combination of joint angles in the passive limb made it difficult to distinguish unequivocally between a limb-based and a joint-based representation. In this study, we decoupled foot position from limb geometry by applying mechanical constraints to individual hindlimb joints in anesthetized cats. We quantified the effect of the joint constraints on limb geometry by analyzing joint-angle covariance in the free and constrained conditions. One type of constraint, a rigid constraint of the knee angle, both changed the covariance pattern and significantly reduced the strength of joint-angle covariance. The other type, an elastic constraint of the ankle angle, changed only the covariance pattern and not its overall strength. We studied the effect of these constraints on the activity in 70 dorsal spinocerebellar tract (DSCT) neurons using a multivariate regression model, with limb axis length and orientation as predictors of neuronal activity. This model also included an experimental condition indicator variable that allowed significant intercept or slope changes in the relationships between foot position parameters and neuronal activity to be determined across conditions. The result of this analysis was that the spatial tuning of 37/70 neurons (53%) was unaffected by the constraints, suggesting that they were somehow able to signal foot position independently from the specific joint angles. We also investigated the extent to which cell activity represented individual joint angles by means of a regression model based on a linear combination of joint angles. A backward elimination of the insignificant predictors determined the set of independent joint angles that best described the neuronal activity for each experimental condition. Finally, by comparing the results of these two approaches, we could determine whether a DSCT neuron represented foot position, specific joint angles, or none of these variables consistently. We found that 10/70 neurons (14%) represented one or more specific joint-angles. The activity of another 27 neurons (39%) was significantly affected by limb geometry changes, but 33 neurons (47%) consistently elaborated a foot position representation in the coordinates of the limb axis. PMID- 10805690 TI - Reference frames for spinal proprioception: kinematics based or kinetics based? AB - This second paper of the series deals with another issue regarding sensorimotor representations in the CNS that has received much attention, namely the relative weighting of kinematic and kinetic representations. The question we address here is the contribution of muscle tension afferent information in dorsal spinocerebellar tract (DSCT) sensory representations of foot position. In five anesthetized cats, we activated major hindlimb muscle groups using electrical stimulation of ventral root filaments while passively positioning of the left hind foot throughout its workspace. In general, as the parameters of the joint angle covariance planes indicated, muscle stimulation did not significantly change hindlimb geometry. We analyzed the effects of the muscle stimulation on DSCT neuronal activity within the framework of a kinematic-based representation of foot position. We used a multivariate regression model described in the companion paper, wherein indicators of the experimental condition were added as firing rate predictors along with the limb axis length and orientation to account for possible effects of muscle stimulation. The results indicated that the response gain of 35/59 neurons studied (59%) was not changed by the muscle activations, although most neurons showed some change in their overall firing level with stimulation of one or more muscles. Most of the neurons responded to pseudorandom stimulation of the same muscle groups with complex temporal patterns of activity. For a subpopulation of 42 neurons, we investigated the extent to which their representation of foot position was affected by a rigid constraint of the knee joint and at least one type of muscle stimulation. Although they could be divided into four subgroups based on significance level cutoffs for the constraint or stimulation effect, these effects were in fact quite distributed. However, when we examined the preferred directions of spatial tuning relative to the limb axis position, we found it was unchanged by muscle stimulation for most cells. Even in those cases in which response gain was altered by muscle stimulation, the cell's preferred direction generally was unaltered. The invariance of preferred direction with muscle stimulation lead us to the conclusion that the reference frame for DSCT coding may be based primarily on limb kinematics. PMID- 10805691 TI - Rate of quantal excitation to a retinal ganglion cell evoked by sensory input. AB - To determine the rate and statistics of light-evoked transmitter release from bipolar synapses, intracellular recordings were made from ON-alpha ganglion cells in the periphery of the intact, superfused, cat retina. Sodium channels were blocked with tetrodotoxin to prevent action potentials. A light bar covering the receptive field center excited the bipolar cells that contact the alpha cell and evoked a transient then a sustained depolarization. The sustained depolarization was quantified as change in mean voltage (Deltav), and the increase in voltage noise that accompanied it was quantified as change in voltage variance (Deltasigma(2)). As light intensity increased, Deltav and Deltasigma(2) both increased, but their ratio held constant. This behavior is consistent with Poisson arrival of transmitter quanta at the ganglion cell. The response component attributable to glutamate quanta from bipolar synapses was isolated by application of 6-cyano-7-nitroquinoxaline (CNQX). As CNQX concentration increased, the signal/noise ratio of this response component (Deltav(CNQX)/Deltasigma(CNQX)) held constant. This is also consistent with Poisson arrival and justified the application of fluctuation analysis. Two different methods of fluctuation analysis applied to Deltav(CNQX) and Deltasigma(CNQX) produced similar results, leading to an estimate that a just maximal sustained response was caused by approximately 3,700 quanta s(-1). The transient response was caused by a rate that was no more than 10-fold greater. Because the ON-alpha cell at this retinal locus has approximately 2,200 bipolar synapses, one synapse released approximately 1.7 quanta s(-1) for the sustained response and no more than 17 quanta s(-1) for the transient. Consequently, within the ganglion cell's integration interval, here calculated to be approximately 16 ms, a bipolar synapse rarely releases more than one quantum. Thus for just maximal sustained and transient depolarizations, the conductance modulated by a single bipolar cell synapse is limited to the quantal conductance ( approximately 100 pS at its peak). This helps preserve linear summation of quanta. The Deltav/Deltasigma(2) ratio remained constant even as the ganglion cell's response saturated, which suggested that even at the peak of sensory input, summation remains linear, and that saturation occurs before the bipolar synapse. PMID- 10805692 TI - Horizontal-disparity tuning of neurons in the visual forebrain of the behaving barn owl. AB - Stereovision plays a major role in depth perception of animals having frontally oriented eyes, most notably primates, cats, and owls. Neuronal mechanisms of disparity sensitivity have only been investigated in anesthetized owls so far. In the current study, responses of 160 visual Wulst neurons to static random-dot stereograms (RDS) were recorded via radiotelemetry in awake, fixating barn owls. The majority of neurons (76%) discharged significantly as a function of horizontal disparity in RDS. The distribution of preferred disparities mirrored the behaviorally relevant range of horizontal disparities that owls can exploit for depth vision. Most tuning profiles displayed periodic modulation and could well be fitted with a Gabor function as expected if disparity detectors were implemented according to the disparity energy model. Corresponding to this observation, a continuum of tuning profiles was observed rather than discrete categories. To assess a possible clustering of neurons with similar disparity tuning properties, single units, and multi-unit activity recorded at individual recording sites were compared. Only a minority of neurons were clustered according to their disparity-tuning properties, suggesting that neurons in the visual Wulst are not organized into columns by preferred disparity. To assess whether variable vergence eye movements influenced tuning data, we correlated tuning peak positions on a trial-by-trial basis for units that were recorded simultaneously. The general lack of significant correlation between single-trial peak positions of simultaneously recorded units indicated that vergence, if at all, had only a minor influence on the data. Our study emphasizes the significance of visual Wulst neurons in analyzing stereoscopic depth information and introduces the barn owl as a second model system to study stereopsis in awake, behaving animals. PMID- 10805693 TI - Abeta fibers mediate cutaneous reflexes during human walking. AB - During human gait, transmission of cutaneous reflexes from the foot is controlled specifically according to the phase of the step cycle. These reflex responses can be evoked by nonnociceptive stimuli, and therefore it is thought that the large myelinated and low-threshold Abeta afferent fibers mediate these reflexes. At present, this hypothesis is not yet verified. To test whether Abeta fibers are involved the reflex responses were studied in patients with a sensory polyneuropathy who suffer from a predominant loss of large-myelinated Abeta fibers. The sural nerve of both patients and healthy control subjects was stimulated electrically at a nonnociceptive intensity during the early and late swing phases while they walked on a treadmill. The responses were studied by recording electromyographic (EMG) activity of the biceps femoris (BF) and tibialis anterior (TA) of the stimulated leg. In both phases, large facilitatory responses were observed in the BF of the healthy subjects. These facilitations were reduced significantly in the BF of the patients, indicating that Abeta fibers mediate these reflexes. In TA similar results were obtained. The absolute response magnitude across the two phases was significantly smaller for the patients than for the healthy subjects. The TA responses for the healthy subjects were on average facilitatory during early swing and suppressive during end swing. Both facilitations and suppressions were considerably smaller for the patients, indicating that both types of responses are mediated by Abeta fibers. It is concluded that low-threshold Abeta sensory fibers mediate these reflexes during human gait. The low threshold and the precise phase-dependent control of these responses suggest that these responses are important in the regulation of gait. The loss of such reflex activity may be related to the gait impairments of these patients. PMID- 10805694 TI - Current and voltage clamp studies of the spike medium afterhyperpolarization of hypoglossal motoneurons in a rat brain stem slice preparation. AB - Whole-cell patch clamp recordings were performed on hypoglossal motoneurons (HMs) in a brain stem slice preparation from the neonatal rat. The medium afterhyperpolarization (mAHP) was the only afterpotential always present after single or multiple spikes, making it suitable for studying its role in firing behavior. At resting membrane potential (-68.8 +/- 0.7 mV), mAHP (23 +/- 2 ms rise-time and 150 +/- 10 ms decay) had 9.5 +/- 0.7 mV amplitude, was suppressed in Ca(2+)-free medium or by 100 nM apamin, and reversed at -94 mV membrane potential. These observations suggest that mAHP was due to activation of Ca(2+) dependent, SK-type K(+) channels. Carbachol (10-100 microM) reversibly and dose dependently blocked the mAHP and depolarized HMs (both effects prevented by 10 microM atropine). Similar mAHP block was produced by muscarine (50 microM). In control solution a constant current pulse (1 s) induced HM repetitive firing with small spike frequency adaptation. When the mAHP was blocked by apamin, the same current pulse evoked much higher frequency firing with strong spike frequency adaptation. Carbachol also elicited faster firing and adapting behavior. Voltage clamp experiments demonstrated a slowly deactivating, apamin-sensitive K(+) current (I(AHP)) which could account for the mAHP. I(AHP) reversed at -94 mV membrane potential, was activated by depolarization as short as 1 ms, decayed with a time constant of 154 +/- 9 ms at -50 mV, and was also blocked by 50 microM carbachol. These data suggest that mAHP had an important role in controlling firing behavior as clearly demonstrated after its pharmacological block and was potently modulated by muscarinic receptor activity. PMID- 10805695 TI - D(1) dopamine receptor activation reduces GABA(A) receptor currents in neostriatal neurons through a PKA/DARPP-32/PP1 signaling cascade. AB - Dopamine is a critical determinant of neostriatal function, but its impact on intrastriatal GABAergic signaling is poorly understood. The role of D(1) dopamine receptors in the regulation of postsynaptic GABA(A) receptors was characterized using whole cell voltage-clamp recordings in acutely isolated, rat neostriatal medium spiny neurons. Exogenous application of GABA evoked a rapidly desensitizing current that was blocked by bicuculline. Application of the D(1) dopamine receptor agonist SKF 81297 reduced GABA-evoked currents in most medium spiny neurons. The D(1) dopamine receptor antagonist SCH 23390 blocked the effect of SKF 81297. Membrane-permeant cAMP analogues mimicked the effect of D(1) dopamine receptor stimulation, whereas an inhibitor of protein kinase A (PKA; Rp 8-chloroadenosine 3',5' cyclic monophosphothioate) attenuated the response to D(1) dopamine receptor stimulation or cAMP analogues. Inhibitors of protein phosphatase 1/2A potentiated the modulation by cAMP analogues. Single-cell RT-PCR profiling revealed consistent expression of mRNA for the beta1 subunit of the GABA(A) receptor-a known substrate of PKA-in medium spiny neurons. Immunoprecipitation assays of radiolabeled proteins revealed that D(1) dopamine receptor stimulation increased phosphorylation of GABA(A) receptor beta1/beta3 subunits. The D(1) dopamine receptor-induced phosphorylation of beta1/beta3 subunits was attenuated significantly in neostriata from DARPP-32 mutants. Voltage-clamp recordings corroborated these results, revealing that the efficacy of the D(1) dopamine receptor modulation of GABA(A) currents was reduced in DARPP 32-deficient medium spiny neurons. These results argue that D(1) dopamine receptor stimulation in neostriatal medium spiny neurons reduces postsynaptic GABA(A) receptor currents by activating a PKA/DARPP-32/protein phosphatase 1 signaling cascade targeting GABA(A) receptor beta1 subunits. PMID- 10805696 TI - Primate translational vestibuloocular reflexes. IV. Changes after unilateral labyrinthectomy. AB - The effects of unilateral labyrinthectomy on the properties of the translational vestibuloocular reflexes (trVORs) were investigated in rhesus monkeys trained to fixate near targets. Translational motion stimuli consisted of either steady state lateral and fore-aft sinusoidal oscillations or short-lasting transient displacements. During small-amplitude, steady-state sinusoidal lateral oscillations, a small decrease in the horizontal trVOR sensitivity and its dependence on viewing distance was observed during the first week after labyrinthectomy. These deficits gradually recovered over time. In addition, the vertical response component increased, causing a tilt of the eye velocity vector toward the lesioned side. During large, transient lateral displacements, the deficits were larger and longer lasting. Responses after labyrinthectomy were asymmetric, with eye velocity during movements toward the side of the lesion being more compromised. The most profound effect of the lesions was observed during fore-aft motion. Whereas responses were kinematically appropriate for fixation away from the side of the lesion (e.g., to the left after right labyrinthectomy), horizontal responses were anticompensatory during fixation at targets located ipsilateral to the side of the lesion (e.g., for targets to the right after right labyrinthectomy). This deficit showed little recovery during the 3-mo post-labyrinthectomy testing period. These results suggest that inputs from both labyrinths are important for the proper function of the trVORs, although the details of how bilateral signals are processed and integrated remain unknown. PMID- 10805697 TI - Cerebellar ataxia: torque deficiency or torque mismatch between joints? AB - Prior work has shown that cerebellar subjects have difficulty adjusting for interaction torques that occur during multi-jointed movements. The purpose of this study was to determine whether this deficit is due to a general inability to generate sufficient levels of phasic torque inability or due to an inability to generate muscle torques that predict and compensate for interaction torques. A second purpose was to determine whether reducing the number of moving joints by external mechanical fixation could improve cerebellar subjects' targeted limb movements. We studied control and cerebellar subjects making elbow flexion movements to touch a target under two conditions: 1) a shoulder free condition, which required only elbow flexion, although the shoulder joint was unconstrained and 2) a shoulder fixed condition, where the shoulder joint was mechanically stabilized so it could not move. We measured joint positions of the arm in the sagittal plane and electromyograms (EMGs) of shoulder and elbow muscles. Elbow and shoulder torques were estimated using inverse dynamics equations. In the shoulder free condition, cerebellar subjects made greater endpoint errors (primarily overshoots) than did controls. Cerebellar subjects' overshoot errors were largely due to unwanted flexion at the shoulder. The excessive shoulder flexion resulted from a torque mismatch, where larger shoulder muscle torques were produced at higher rates than would be appropriate for a given elbow movement. In the shoulder fixed condition, endpoint errors of cerebellar subjects and controls were comparable. The improved accuracy of cerebellar subjects was accompanied by reduced shoulder flexor muscle activity. Most of the correct cerebellar trials in the shoulder fixed condition were movements made using only muscles that flex the elbow. Our findings suggest that cerebellar subjects' poor shoulder control is due to an inability to generate muscle torques that predict and compensate for interaction torques, and not due to a general inability to generate sufficient levels of phasic torque. In addition, reducing the number of muscles to be controlled improved cerebellar ataxia. PMID- 10805698 TI - Multiple forms of short-term plasticity at excitatory synapses in rat medial prefrontal cortex. AB - Short-term synaptic plasticity, in particular short-term depression and facilitation, strongly influences neuronal activity in cerebral cortical circuits. We investigated short-term plasticity at excitatory synapses onto layer V pyramidal cells in the rat medial prefrontal cortex, a region whose synaptic dynamic properties have not been systematically examined. Using intracellular and extracellular recordings of synaptic responses evoked by stimulation in layers II/III in vitro, we found that short-term depression and short-term facilitation are similar to those described previously in other regions of the cortex. In addition, synapses in the prefrontal cortex prominently express augmentation, a longer lasting form of short-term synaptic enhancement. This consists of a 40-60% enhancement of synaptic transmission which lasts seconds to minutes and which can be induced by stimulus trains of moderate duration and frequency. Synapses onto layer III neurons in the primary visual cortex express substantially less augmentation, indicating that this is a synapse-specific property. Intracellular recordings from connected pairs of layer V pyramidal cells in the prefrontal cortex suggest that augmentation is a property of individual synapses that does not require activation of multiple synaptic inputs or neuromodulatory fibers. We propose that synaptic augmentation could function to enhance the ability of a neuronal circuit to sustain persistent activity after a transient stimulus. This idea is explored using a computer simulation of a simplified recurrent cortical network. PMID- 10805699 TI - Contextual modulation of substantia nigra pars reticulata neurons. AB - Neurons in the substantia nigra pars reticulata (SNr) are known to encode saccadic eye movements within some, but not all, behavioral contexts. However, the precise contextual factors that effect the modulations of nigral activity are still uncertain. To further examine the effect of behavioral context on the SNr, we recorded the activity of 72 neurons while monkeys made saccades during a delayed saccade task and during periods of free viewing. We quantified and compared the movement fields of each neuron for saccades made under three different conditions: 1) spontaneous saccades, which shifted gaze during periods of free viewing when no stimuli were presented and no reinforcements were delivered; 2) fixational saccades, which brought gaze into alignment with a fixation target at the start of a delayed saccade trial, were necessary for trial completion, but were not directly followed by reinforcement; and 3) terminal saccades, which brought gaze into alignment with a visual target at the end of a delayed saccade trial and were directly followed by reinforcement. For three of the four SNr neuron classes, saccade-related modulations were only present before terminal saccades. For the fourth class, discrete pausers, saccade-related modulations were substantially larger for terminal saccades than for fixational saccades, and modulations were absent for spontaneous saccades. These results and other recent work on the basal ganglia suggest that some saccade-related signals in the SNr may be influenced by the reinforcement associated with a particular saccadic eye movement. PMID- 10805700 TI - Encoding of location and intensity of noxious indentation into rat skin by spatial populations of cutaneous mechano-nociceptors. AB - The ability of a spatial population of cutaneous, Adelta, and C mechano nociceptors to encode the location and intensity of a noxious, cutaneous indentation was examined using an isolated preparation in a rat model. Skin and its intact innervation were harvested from the medial thigh of the rat hindlimb and placed in a dish, with the corium side down, containing synthetic interstitial fluid. The margins of the skin were coupled to an apparatus that could stretch and apply compression to the skin. The skin was suspended on top of a deformable platform whose bulk, nonlinear, compressive compliance emulated that found in vivo. The isolated preparation facilitated examination of the spatial population response by eliminating the nonlinear geometry and inhomogeneous compressive compliance present in-vivo. Spatial population responses (SPR) were formed from recordings of single neurons that were stimulated by compressing the skin with an indenter (flat cylinder, 3-mm diam) at discrete intervals from the center of their receptive fields. SPR were composed of the neural responses (z axis) at each indentation location (x, y plane), and were analyzed quantitatively using nonlinear regression to fit an equation of a Gaussian surface. Both Adelta and C SPR accurately encoded the location and intensity of noxious indentation. The intensity of the stimulus was encoded in the peak neural response of the SPR, which had a nonlinear relationship to the compressive force. The location of the stimulus was encoded in the x, y position of the peak of the SPR. The position of the peak remained constant with increasing magnitudes of compressive force. The overall form of the SPR also remained constant with changes of compressive load, suggesting a possible role for encoding in the SPR some aspects of shape of a noxious stimulus. PMID- 10805701 TI - Selective spatial attention in vision and touch: unimodal and multimodal mechanisms revealed by PET. AB - Two positron-emission tomography (PET) experiments explored the neural basis of selective spatial attention in vision and touch, testing for modality-specific versus multimodal activations due to attended side. In the first study, either light flashes or finger vibrations were presented bilaterally. Twelve healthy volunteers were scanned while sustaining covert attention on the left or right hemifield within each modality. The main effect for attending right minus left, across both modalities, revealed bimodal spatial attention effects in the left intraparietal sulcus and left occipitotemporal junction. Modality-specific attentional effects (again, for attending right vs. left) were found in the left superior occipital gyrus for vision, and left superior postcentral gyrus for touch. No significant activations were seen for attending left minus right. The second study presented only tactile stimuli, manipulating whether the eyes were open or closed, and including passive stimulation and rest baselines. The unimodal activation for tactile spatial attention in the left superior postcentral gyrus was replicated. The bimodal activation of the left intraparietal sulcus observed in the first study was now found for touch, but only when the eyes were open (hands visible), apparently confirming its multimodal nature. These results reveal mechanisms of sustained spatial attention operating at both modality-specific and multimodal levels. PMID- 10805702 TI - Retroambiguus projections to the cutaneus trunci motoneurons may form a pathway in the central control of mating. AB - Our laboratory has proposed that the nucleus retroambiguus (NRA) generates the specific motor performance displayed by female cats during mating and that it uses direct pathways to the motoneurons of the lower limb muscles involved in this activity. In the hamster a similar NRA-projection system could generate the typical female mating posture, which is characterized by lordosis of the back as well as elevation of the tail. The present study attempted to determine whether this elevation of the tail is also part of the NRA-mating control system. The basic assumption was that elevation of the tail is a function of the cutaneous trunci muscle (CTM), which was verified by bilateral tetanic stimulation of the lateral thoracic nerves innervating the CTM. It resulted in upward movement of the tail to a position similar to the tail-up position during the lordosis posture. Retrograde tracing results showed that CTM motoneurons are located in the ventral and ventrolateral part of the C(7)-C(8) ventral horn, those innervating the tail region ventrolateral to those innervating the axillary region. Anterograde tracing studies showed that NRA fibers terminate bilaterally in both parts of the CTM motoneuronal cell groups. Electron microscopical studies revealed that labeled NRA terminals make monosynaptic contacts with retrogradely labeled dendrites of CTM motoneurons. Almost all of these terminal profiles had asymmetric synapses and contained spherical vesicles, which suggests an excitatory function. The observation that 15% of the labeled NRA terminals make more than one synaptic contact with a retrogradely labeled CTM motoneuronal dendrite within the same section indicates how powerful the NRA-CTM projection is. The results indicate that during mating the NRA not only could generate the lordosis posture but also the elevation of the tail. PMID- 10805703 TI - Coupled oscillator model of the dopaminergic neuron of the substantia nigra. AB - Calcium imaging using fura-2 and whole cell recording revealed the effective location of the oscillator mechanism on dopaminergic neurons of the substantia nigra, pars compacta, in slices from rats aged 15-20 days. As previously reported, dopaminergic neurons fired in a slow rhythmic single spiking pattern. The underlying membrane potential oscillation survived blockade of sodium currents with TTX and was enhanced by blockade of voltage-sensitive potassium currents with TEA. Calcium levels increased during the subthreshold depolarizing phase of the membrane potential oscillation and peaked at the onset of the hyperpolarizing phase as expected if the pacemaker potential were due to a low threshold calcium current and the hyperpolarizing phase to calcium-dependent potassium current. Calcium oscillations were synchronous in the dendrites and soma and were greater in the dendrites than in the soma. Average calcium levels in the dendrites overshot steady-state levels and decayed over the course of seconds after the oscillation was resumed after having been halted by hyperpolarizing currents. Average calcium levels in the soma increased slowly, taking many cycles to achieve steady state. Voltage clamp with calcium imaging revealed the voltage dependence of the somatic calcium current without the artifacts of incomplete spatial voltage control. This showed that the calcium current had little or no inactivation and was half-maximal at -40 to -30 mV. The time constant of calcium removal was measured by the return of calcium to resting levels and depended on diameter. The calcium sensitivity of the calcium-dependent potassium current was estimated by plotting the slow tail current against calcium concentration during the decay of calcium to resting levels at -60 mV. A single compartment model of the dopaminergic neuron consisting of a noninactivating low threshold calcium current, a calcium-dependent potassium current, and a small leak current reproduced most features of the membrane potential oscillations. The same currents much more accurately reproduced the calcium transients when distributed uniformly along a tapering cable in a multicompartment model. This model represented the dopaminergic neuron as a set of electrically coupled oscillators with different natural frequencies. Each frequency was determined by the surface area to volume ratio of the compartment. This model could account for additional features of the dopaminergic neurons seen in slices, such as slow adaptation of oscillation frequency and may produce irregular firing under different coupling conditions. PMID- 10805704 TI - Thyroid hormone deficiency before the onset of hearing causes irreversible damage to peripheral and central auditory systems. AB - Both a genetic or acquired neonatal thyroid hormone (TH) deficiency may result in a profound mental disability that is often accompanied by deafness. The existence of various TH-sensitive periods during inner ear development and general success of delayed, corrective TH treatment was investigated by treating pregnant and lactating rats with the goitrogen methimazole (MMI). We observed that for the establishment of normal hearing ability, maternal TH, before fetal thyroid gland function on estrus days 17-18, is obviously not required. Within a crucial time between the onset of fetal thyroid gland function and the onset of hearing at postnatal day 12 (P12), any postponement in the rise of TH-plasma levels, as can be brought about by treating lactating mothers with MMI, leads to permanent hearing defects of the adult offspring. The severity of hearing defects that were measured in 3- to 9-mo-old offspring could be increased with each additional day of TH deficiency during this critical period. Unexpectedly, the active cochlear process, assayed by distortion product otoacoustic emissions (DPOAE) measurements, and speed of auditory brain stem responses, which both until now were not thought to be controlled by TH, proved to be TH-dependent processes that were damaged by a delay of TH supply within this critical time. In contrast, no significant differences in the gross morphology and innervation of the organ of Corti or myelin gene expression in the auditory system, detected as myelin basic protein (MBP) and proteolipid protein (PLP) mRNA using Northern blot approach, were observed when TH supply was delayed for few days. These classical TH dependent processes, however, were damaged when TH supply was delayed for several weeks. These surprising results may suggest the existence of different TH dependent processes in the auditory system: those that respond to corrective TH supply (e.g., innervation and morphogenesis of the organ of Corti) and those that do not, but require T3 activity during a very tight time window (e.g. , active cochlear process, central processes). PMID- 10805705 TI - Cortical responses to thermal pain depend on stimulus size: a functional MRI study. AB - Cortical activity patterns to thermal painful stimuli of two different sizes were examined in normal volunteers using functional magnetic resonance imaging (fMRI). Seven right-handed subjects were studied when the painful stimulus applied to the right hand fingers covered either 1,074-mm(2)-area large stimulator or 21-mm(2) area small stimulator. Stimulus temperatures were adjusted to give rise to equivalent moderately painful ratings. fMRI signal increases and decreases were determined for the contralateral parietal and motor areas. When the overall activity in these regions was compared across subjects, increased fMRI activity was observed over more brain volume with the larger stimulator, whereas decreased fMRI activity was seen in more brain volume for the smaller stimulator. The individual subject and group-averaged activity patterns indicated regional specific differences in increased and decreased fMRI activity. The small stimulator resulted in decreased fMRI responses throughout the upper body representation in both primary somatosensory and motor cortices. In contrast, no decreased fMRI signals were seen in the secondary somatosensory cortex and in the insula. In another seven volunteers, the effects of the size of the thermal painful stimulus on vibrotactile thresholds were examined psychophysically. Painful stimuli were delivered to the fingers and vibrotactile thresholds were measured on the arm just distal to the elbow. Consistent with the fMRI results in the primary somatosensory cortex, painful thermal stimuli using the small stimulator increased vibrotactile thresholds on the forearm, whereas similarly painful stimuli using the large stimulator had no effect on forearm vibrotactile thresholds. These results are discussed in relation to the cortical dynamics for pain perception and in relation to the center-surround organization of cortical neurons. PMID- 10805706 TI - Topographic analysis of epidural pure-tone-evoked potentials in gerbil auditory cortex. AB - This study investigated the tonotopic organization of pure-tone-evoked middle latency auditory evoked potentials (MAEPs) recorded at the auditory cortical surface in unanesthetized gerbils. Multielectrode array recording and multiple linear regression analysis of the MAEP demonstrated different degrees of tonotopic organization of early and late MAEP components. The early MAEP components P1 and N1 showed focal topography and clear dependence in location and size of cortical area covered on pure-tone frequency. The later components P2 and N2 showed a widespread topography which was largely unaffected in location and size of cortical area covered by pure-tone frequency. These results allow delimitation of the neural generators of the early and late MAEP components in terms of the spectral properties of functionally defined neural populations. PMID- 10805707 TI - Responses to rare visual target and distractor stimuli using event-related fMRI. AB - Previous studies have found that the P300 or P3 event-related potential (ERP) component is useful in the diagnosis and treatment of many disorders that influence CNS function. However, the anatomic locations of brain regions involved in this response are not precisely known. In the present event-related functional magnetic resonance imaging (fMRI) study, methods of stimulus presentation, data acquisition, and data analysis were optimized for the detection of brain activity in response to stimuli presented in the three-stimulus oddball task. This paradigm involves the interleaved, pseudorandom presentation of single block letter target and distractor stimuli that previously were found to generate the P3b and P3a ERP subcomponents, respectively, and frequent standard stimuli. Target stimuli evoked fMRI signal increases in multiple brain regions including the thalamus, the bilateral cerebellum, and the occipital-temporal cortex as well as bilateral superior, medial, inferior frontal, inferior parietal, superior temporal, precentral, postcentral, cingulate, insular, left middle temporal, and right middle frontal gyri. Distractor stimuli evoked an fMRI signal change bilaterally in inferior anterior cingulate, medial frontal, inferior frontal, and right superior frontal gyri, with additional activity in bilateral inferior parietal lobules, lateral cerebellar hemispheres and vermis, and left fusiform, middle occipital, and superior temporal gyri. Significant variation in the amplitude and polarity of distractor-evoked activity was observed across stimulus repetitions. No overlap was observed between target- and distractor-evoked activity. These event-related fMRI results shed light on the anatomy of responses to target and distractor stimuli that have proven useful in many ERP studies of healthy and clinically impaired populations. PMID- 10805708 TI - Parietal neurons represent surface orientation from the gradient of binocular disparity. AB - In order to elucidate the neural mechanisms involved in the perception of the three-dimensional (3D) orientation of a surface, we trained monkeys to discriminate the 3D orientation of a surface from binocular disparity cues using a Go/No-go type delayed-matching-to-sample (DMTS) task and examined the properties of the surface-orientation-selective (SOS) neurons. We recorded 57 SOS neurons from the caudal part of the lateral bank of the intraparietal sulcus (area CIP) of three hemispheres of two Japanese monkeys (Macaca fuscata). We tested 29 of 57 SOS neurons using the square plate of a solid figure stereogram (SFS) and random-dot stereogram (RDS) without perspective cues; almost all of the tested neurons (28/29) showed surface orientation selectivity for the SFS and/or the RDS without perspective cues. Eight of these 28 neurons (28.6%) showed selectivity for both the RDS and SFS, 7 (25.0%) were dominantly selective for the RDS, and 13 (46.4%) were dominantly selective for the SFS. These results suggest that neurons that show surface orientation tuning for the RDS without perspective cues compute surface orientation from the gradient of the binocular disparity given by the random-dot across the surface. On the other hand, neurons that show surface orientation tuning for the SFS without perspective cues may represent surface orientation primarily from the gradient of the binocular disparity along the contours. In conclusion, the SOS neurons in the area CIP are likely to operate higher order processing of disparity signals for surface perception by integrating the input signals from many disparity-sensitive neurons with different disparity tuning. PMID- 10805709 TI - Plasticity in the distribution of the red nucleus output to forearm muscles after unilateral lesions of the pyramidal tract. AB - It has been hypothesized that the magnocellular red nucleus (RNm) contributes to compensation for motor impairments associated with lesions of the pyramidal tract. To test this hypothesis, we used stimulus triggered averaging (StTA) of electromyographic (EMG) activity to characterize changes in motor output from the red nucleus after lesions of the pyramidal tract. Three monkeys were trained to perform a reach and prehension task. EMG activity was recorded from 11 forearm muscles including one elbow, five wrist, and five digit muscles. Microstimulation (20 microA at 20 Hz) was delivered throughout the movement task to compute StTAs. Two monkeys served as controls. In a third monkey, 65% of the left pyramidal tract had been destroyed by an electrolytic lesion method five years before recording. The results demonstrate a clear pattern of postlesion reorganization in red nucleus-mediated output effects on forearm muscles. The normally prominent extensor preference in excitatory output from the RNm (92% in extensors) was greatly diminished in the lesioned monkey (59%). Similarly, suppression effects, which are normally much more prominent in flexor than in extensor muscles (90% in flexors), were also more evenly distributed after recovery from pyramidal tract lesions. Because of the limited excitatory output from the RNm to flexor muscles that normally exists, loss of corticospinal output would leave control of flexors particularly weak. The changes in RNm organization reported in this study would help restore function to flexor muscles. These results support the hypothesis that the RNm is capable of reorganization that contributes to the recovery of forelimb motor function after pyramidal tract lesions. PMID- 10805710 TI - Thalamic and cortical contributions to neural plasticity after limb amputation. AB - Little is known about the substrates for the large-scale shifts in the cortical representation produced by limb amputation. Subcortical changes likely contribute to the cortical remodeling, yet there is little data regarding the extent and pattern of reorganization in thalamus after such a massive deafferentation. Moreover, the relationship between changes in thalamus and in cortex after injuries of this nature is virtually unexplored. Multiunit microelectrode maps were made in the somatosensory thalamus and cortex of two monkeys that had long standing, accidental forelimb amputations. In the deprived portion of the ventroposterior nucleus of the thalamus (VP), where stimulation to the hand would normally activate neurons, new receptive fields had emerged. At some recording sites within the deprived zone of VP, neurons responded to stimulation of the remaining stump of the arm and at other sites neurons responded to stimulation of both the stump and the face. This same overall pattern of reorganization was present in the deprived hand representation of cortical area 3b. Thus thalamic changes produced by limb amputation appear to be an important substrate of cortical reorganization. However, a decrease in the frequency of abnormal stump/face fields in area 3b compared with VP and a reduction in the size of the fields suggests that cortical mechanisms of plasticity may refine the information relayed from thalamus. PMID- 10805711 TI - Time-dependent changes in input resistance of rat hypoglossal motoneurons associated with whole-cell recording. AB - The effect of cellular dialysis associated with whole-cell recording was studied in 24 developing hypoglossal motoneurons in a rat brainstem slice preparation. In all cases, establishing whole-cell continuity with the electrode solution resulted in an increase in the input resistance measured in current clamp. The mean magnitude of this increase was 39.7% and the time course of the maximum effect was quite variable. There was no correlation found between the time to maximum effect and the magnitude of the increase in resistance. These data indicate that the passive membrane properties are not constant during whole-cell recording in mammalian motoneurons. PMID- 10805712 TI - Phasic lung inflation shortens inspiration and respiratory period in the lung attached neonate rat brain stem spinal cord. AB - In intact mammals, lung inflation during inspiration terminates inspiration (Breuer-Hering inspiratory reflex, BHI) and the presence of lung afferents increases respiratory frequency. To test whether these responses could be obtained in vitro, a neonate rat brain stem/spinal cord preparation retaining the lungs and their vagal innervation was used. It was found that 1) the BHI could be replicated in vitro, 2) phasic lung inflation during inspiration caused increased respiratory frequency with declining efficacy as inflation delay increased, and 3) increased respiratory frequency did not require inspiratory shortening. PMID- 10805713 TI - Equilibrium potential of GABA(A) current and implications for rebound burst firing in rat subthalamic neurons in vitro. AB - Reciprocally connected glutamatergic subthalamic and GABAergic globus pallidus neurons have recently been proposed to act as a generator of low-frequency oscillatory activity in Parkinson's disease. To determine whether GABA(A) receptor-mediated synaptic potentials could theoretically generate rebound burst firing in subthalamic neurons, a feature that is central to the proposed oscillatory mechanism, we determined the equilibrium potential of GABA(A) current (E(GABA(A))) and the degree of hyperpolarization required for rebound firing using perforated-patch recording. In the majority of neurons that fired rebounds, E(GABA(A)) was equal to or more hyperpolarized than the hyperpolarization required for rebound burst firing. These data suggest that synchronous activity of pallidal inputs could underlie rhythmic bursting activity of subthalamic neurons in Parkinson's disease. PMID- 10805714 TI - Calcium-independent afterdepolarization regulated by serotonin in anterior thalamus. AB - Previous studies have identified an afterdepolarization (ADP) in thalamocortical neurons that is mediated by an upregulation of the hyperpolarization-activated current I(h). This ADP has been suggested to play a key role in the generation of spindle oscillations. In the lateral geniculate nucleus, upregulation of I(h) has been shown to be signaled by a rise in intracellular calcium leading to the activation of adenylate cyclase and formation of cAMP. However, it is unclear how generalizable this mechanism is to other thalamic nuclei. We have used whole cell recording to examine the electrophysiological properties of neurons of the anterodorsal thalamic nucleus, a nucleus thought not to undergo spindle oscillations. We now report that cells in this nucleus also display an ADP mediated by I(h). Surprisingly, the ADP and the underlying upregulation of I(h) persisted even after buffering intracellular calcium and blocking calcium influx. These results indicate that, in neurons of the anterodorsal thalamic nucleus, an I(h)-mediated ADP can occur through a mechanism that does not involve a rise in intracellular calcium. We next examined the possibility that this calcium independent ADP might be modulated by serotonin. Serotonin produced a robust enhancement in the amplitude of the ADP even after strong buffering of intracellular calcium and blockade of calcium channels. These results indicate that neurons of the anterodorsal thalamic nucleus display a calcium-independent, I(h)-mediated ADP and that this ADP is a target for regulation by serotonin. These findings identify a novel mechanism by which serotonin can regulate neuronal excitability. PMID- 10805715 TI - Site independence of EPSP time course is mediated by dendritic I(h) in neocortical pyramidal neurons. AB - Neocortical layer 5 pyramidal neurons possess long apical dendrites that receive a significant portion of the neurons excitatory synaptic input. Passive neuronal models indicate that the time course of excitatory postsynaptic potentials (EPSPs) generated in the apical dendrite will be prolonged as they propagate toward the soma. EPSP propagation may, however, be influenced by the recruitment of dendritic voltage-activated channels. Here we investigate the properties and distribution of I(h) channels in the axon, soma, and apical dendrites of neocortical layer 5 pyramidal neurons, and their effect on EPSP time course. We find a linear increase (9 pA/100 microm) in the density of dendritic I(h) channels with distance from soma. This nonuniform distribution of I(h) channels generates site independence of EPSP time course, such that the half-width at the soma of distally generated EPSPs (up to 435 microm from soma) was similar to somatically generated EPSPs. As a corollary, a normalization of temporal summation of EPSPs was observed. The site independence of somatic EPSP time course was found to collapse after pharmacological blockade of I(h) channels, revealing pronounced temporal summation of distally generated EPSPs, which could be further enhanced by TTX-sensitive sodium channels. These data indicate that an increasing density of apical dendritic I(h) channels mitigates the influence of cable filtering on somatic EPSP time course and temporal summation in neocortical layer 5 pyramidal neurons. PMID- 10805716 TI - Hippocampus-entorhinal cortex loop and seizure generation in the young rodent limbic system. AB - Application of the convulsant 4-aminopyridine (4AP, 50 microM) to adult mouse combined hippocampus-entorhinal cortex (EC) slices induces interictal and ictal discharges originating from CA3 and EC respectively. In this model of limbic seizures, ictal discharges disappear over time and are reestablished after Schaffer collateral cut, a procedure that blocks interictal propagation from CA3 to EC. Here we tested whether this form of network plasticity is operant in hippocampus-EC slices obtained from young (10-25 day-old) mice. In these experiments 4AP elicited interictal (duration = 100-250 ms; interval = 0.7 +/- 0.2 s, mean +/- SD, n = 20) and ictal (duration = 267 +/- 37 s; interval = 390 +/ 37 s, n = 20) discharges in both CA3 and EC. However, in young mouse slices the ictal events occurred throughout the experiment, whereas Schaffer collateral cut abolished CA3-driven interictal discharges in EC without influencing ictal activity (n = 10). Perforant path lesion prevented the spread of EC-driven ictal events to CA3, where interictal and short ictal discharges (duration = 32 +/- 11 s; interval = 92 +/- 9.7 s, n = 8) continued to occur. Hence, two independent forms of ictal activity were seen in CA3 and in EC after separation of these structures. In intact hippocampus-EC slices, ictal discharges were reduced by an N-methyl-D-aspartate receptor antagonist (n = 10). Under these conditions, Schaffer collateral cut abolished ictal activity in EC, not in CA3 (n = 6). Thus the young mouse hippocampus-EC loop has different properties as compared with adult tissue. These differences, which include the inability of hippocampal outputs to control ictal discharge generation in EC and the ability of the loop to sustain ictal activity, may contribute to the low-seizure threshold seen in young individuals. PMID- 10805717 TI - Control of the wrist in three-joint arm movements to multiple directions in the horizontal plane. AB - In a reaching movement, the wrist joint is subject to inertial effects from proximal joint motion. However, precise control of the wrist is important for reaching accuracy. Studies of three-joint arm movements report that the wrist joint moves little during point-to-point reaches, but muscle activities and kinetics have not yet been described across a range of movement directions. We hypothesized that to minimize wrist motion, muscle torques at the wrist must perfectly counteract inertial effects arising from proximal joint motion. Subjects were given no instructions regarding joint movement and were observed to keep the wrist nearly motionless during center-out reaches to directions throughout the horizontal plane. Consistent with this, wrist muscle torques exactly mirrored interaction torques, in contrast to muscle torques at proximal joints. These findings suggest that in this reaching task the nervous system chooses to minimize wrist motion by anticipating dynamic inertial effects. The wrist muscle torques were associated with a direction-dependent choice of muscles, also characterized by initial reciprocal activation rather than initial coactivation to stiffen the wrist joint. In a second experiment, the same pattern of muscle activities persisted even after many trials reaching with the wrist joint immobilized. These results, combined with similar features at the three joints, such as cosine-like tuning of muscle torques and of muscle onsets across direction, suggest that the nervous system uses similar rules for muscles at each joint, as part of one plan for the arm during a point-to-point reach. PMID- 10805718 TI - Modulation of pacemaker activity by salmon gonadotropin-releasing hormone (sGnRH) in terminal nerve (TN)-GnRH neurons. AB - The terminal nerve (TN)-gonadotropin-releasing hormone (GnRH) neurons project widely in the brain instead of the pituitary and show endogenous pacemaker activity that is dependent on the physiological conditions of the animal. We suggest that the TN-GnRH system may act as a putative neuromodulator that is involved in the regulation of many long-lasting changes in the animal's behavior. In the present study, we find that the pacemaker activity of TN-GnRH neurons is modulated by salmon GnRH (sGnRH), which is the same molecular species of GnRH peptide produced by TN-GnRH neurons themselves. Bath application of sGnRH (2-200 nM) transiently decreased (early phase) and then subsequently increased (late phase) the frequency of pacemaker activity of TN-GnRH neurons in a dose-dependent manner. These biphasic changes of pacemaker activities were suppressed by intracellular application of guanosin 5'-0-(2-thiodi-phosphate) (GDP-beta-S). The results suggest that G-protein coupled receptors are present on the cell surface and play a triggering role in modulating the frequency of pacemaker activities in TN-GnRH neurons. Because the TN-GnRH neurons make tight cell clusters with no intervening glial cells, it may be further suggested that GnRH released from GnRH neurons regulates the activities of their own (autocrine) and/or neighboring GnRH neurons (paracrine). PMID- 10805719 TI - Somatic mRNA turnover mutants implicate tristetraprolin in the interleukin-3 mRNA degradation pathway. AB - Control of mRNA stability is critical for expression of short-lived transcripts from cytokines and proto-oncogenes. Regulation involves an AU-rich element (ARE) in the 3' untranslated region (3'UTR) and cognate trans-acting factors thought to promote either degradation or stabilization of the mRNA. In this study we present a novel approach using somatic cell genetics designed to identify regulators of interleukin-3 (IL-3) mRNA turnover. Mutant cell lines were generated from diploid HT1080 cells transfected with a reporter construct containing green fluorescent protein (GFP) linked to the IL-3 3'UTR. GFP was expressed at low levels due to rapid decay of the mRNA. Following chemical mutagenesis and selection of GFP overexpressing cells, we could isolate three mutant clones (slowA, slowB, and slowC) with a specific, trans-acting defect in IL-3 mRNA degradation, while the stability of IL-2 and tumor necrosis factor alpha reporter transcripts was not affected. Somatic cell fusion experiments revealed that the mutants are genetically recessive and form two complementation groups. Expression of the tristetraprolin gene in both groups led to reversion of the mutant phenotype, thereby linking this gene to the IL-3 mRNA degradation pathway. The genetic approach described here should allow identification of the defective functions by gene transfer and is also applicable to the study of other mRNA turnover pathways. PMID- 10805720 TI - Induction of hTERT expression and telomerase activity by estrogens in human ovary epithelium cells. AB - In mammals, molecular mechanisms and factors involved in the tight regulation of telomerase expression and activity are still largely undefined. In this study, we provide evidence for a role of estrogens and their receptors in the transcriptional regulation of hTERT, the catalytic subunit of human telomerase and, consequently, in the activation of the enzyme. Through a computer analysis of the hTERT 5'-flanking sequences, we identified a putative estrogen response element (ERE) which was capable of binding in vitro human estrogen receptor alpha (ERalpha). In vivo DNA footprinting revealed specific modifications of the ERE region in ERalpha-positive but not ERalpha-negative cells upon treatment with 17beta-estradiol (E2), indicative of estrogen-dependent chromatin remodelling. In the presence of E2, transient expression of ERalpha but not ERbeta remarkably increased hTERT promoter activity, and mutation of the ERE significantly reduced this effect. No telomerase activity was detected in human ovary epithelial cells grown in the absence of E2, but the addition of the hormone induced the enzyme within 3 h of treatment. The expression of hTERT mRNA and protein was induced in parallel with enzymatic activity. This prompt estrogen modulation of telomerase activity substantiates estrogen-dependent transcriptional regulation of the hTERT gene. The identification of hTERT as a target of estrogens represents a novel finding which advances the understanding of telomerase regulation in hormone dependent cells and has implications for a potential role of hormones in their senescence and malignant conversion. PMID- 10805721 TI - Analysis of ku80-mutant mice and cells with deficient levels of p53. AB - Absence of Ku80 results in increased sensitivity to ionizing radiation, defective lymphocyte development, early onset of an age-related phenotype, and premature replicative senescence. Here we investigate the role of p53 on the phenotype of ku80-mutant mice and cells. Reducing levels of p53 increased the cancer incidence for ku80(-/-) mice. About 20% of ku80(-/-) p53(+/-) mice developed a broad spectrum of cancer by 40 weeks and all ku80(-/-) p53(-/-) mice developed pro-B cell lymphoma by 16 weeks. Reducing levels of p53 rescued populations of ku80(-/ ) cells from replicative senescence by enabling spontaneous immortalization. The double-mutant cells are impaired for the G(1)/S checkpoint due to the p53 mutation and are hypersensitive to gamma-radiation and reactive oxygen species due to the Ku80 mutation. These data show that replicative senescence is caused by a p53-dependent cell cycle response to damaged DNA in ku80(-/-) cells and that p53 is essential for preventing very early onset of pro-B-cell lymphoma in ku80( /-) mice. PMID- 10805722 TI - Granzyme B short-circuits the need for caspase 8 activity during granule-mediated cytotoxic T-lymphocyte killing by directly cleaving Bid. AB - Cytotoxic T lymphocytes (CTL) can trigger an apoptotic signal through the Fas receptor or by the exocytosis of granzyme B and perforin. Caspase activation is an important component of both pathways. Granzyme B, a serine proteinase contained in granules, has been shown to proteolytically process and activate members of the caspase family in vitro. In order to gain an understanding of the contributions of caspases 8 and 3 during granule-induced apoptosis in intact cells, we have used target cells that either stably express the rabbitpox virus encoded caspase inhibitor SPI-2 or are devoid of caspase 3. The overexpression of SPI-2 in target cells significantly inhibited DNA fragmentation, phosphatidylserine externalization, and mitochondrial disruption during Fas mediated cell death. In contrast, SPI-2 expression in target cells provided no protection against granzyme-mediated apoptosis, mitochondrial collapse, or cytolysis, leading us to conclude that SPI-2-inhibited caspases are not an essential requirement for the granzyme pathway. Caspase 3-deficient MCF-7 cells were found to be resistant to CTL-mediated DNA fragmentation but not to CTL mediated cytolysis and loss of the mitochondrial inner membrane potential. Furthermore, we demonstrate that granzyme B directly cleaves the proapoptotic molecule Bid, bypassing the need for caspase 8 activation of Bid. These results provide evidence for a two-pronged strategy for mediating target cell destruction and provide evidence of a direct link between granzyme B activity, Bid cleavage, and caspase 3 activation in whole cells. PMID- 10805723 TI - Hierarchy of S-phase-promoting factors: yeast Dbf4-Cdc7 kinase requires prior S phase cyclin-dependent kinase activation. AB - In all eukaryotes, the initiation of DNA synthesis requires the formation of prereplicative complexes (pre-RCs) on replication origins, followed by their activation by two S-T protein kinases, an S-phase cyclin-dependent kinase (S-CDK) and a homologue of yeast Dbf4-Cdc7 kinase (Dbf4p-dependent kinase [DDK]). Here, we show that yeast DDK activity is cell cycle regulated, though less tightly than that of the S-CDK Clb5-Cdk1, and peaks during S phase in correlation with Dbf4p levels. Dbf4p is short-lived throughout the cell cycle, but its instability is accentuated during G(1) by the anaphase-promoting complex. Downregulating DDK activity is physiologically important, as joint Cdc7p and Dbf4p overexpression is lethal. Because pre-RC formation is a highly ordered process, we asked whether S CDK and DDK need also to function in a specific order for the firing of origins. We found that both kinases are activated independently, but we show that DDK can perform its function for DNA replication only after S-CDKs have been activated. Cdc45p, a protein needed for initiation, binds tightly to chromatin only after S CDK activation (L. Zou and B. Stillman, Science 280:593-596, 1998). We show that Cdc45p is phosphorylated by DDK in vitro, suggesting that it might be one of DDK's critical substrates after S-CDK activation. Linking the origin-bound DDK to the tightly regulated S-CDK in a dependent sequence of events may ensure that DNA replication initiates only at the right time and place. PMID- 10805724 TI - Three yeast proteins related to the human candidate tumor suppressor p33(ING1) are associated with histone acetyltransferase activities. AB - Three Saccharomyces cerevisiae proteins (Yng1/YOR064c, Yng2/YHR090c, and Pho23) and two Schizosaccharomyces pombe proteins (Png1/CAA15917 and Png2/CAA21250) share significant sequence identity with the human candidate tumor suppressor p33(ING1) in their C-terminal regions. The homologous regions contain PHD finger domains which have been implicated in chromatin-mediated transcriptional regulation. We show that GFP-Yng2, like human Ing1, is localized in the nucleus. Deletion of YNG2 results in several phenotypes, including an abnormal multibudded morphology, an inability to utilize nonfermentable carbon sources, heat shock sensitivity, slow growth, temperature sensitivity, and sensitivity to caffeine. These phenotypes are suppressed by expression of either human Ing1 or S. pombe Png1, suggesting that the yeast and human proteins are functionally conserved. Yng1- and Pho23-deficient cells also share some of these phenotypes. We demonstrated by yeast two-hybrid and coimmunoprecipitation tests that Yng2 interacts with Tra1, a component of histone acetyltransferase (HAT) complexes. We further demonstrated by coimmunoprecipitation that HA-Yng1, HA-Yng2, HA-Pho23, and HA-Ing1 are associated with HAT activities in yeast. Genetic and biochemical evidence indicate that the Yng2-associated HAT is Esa1, suggesting that Yng2 is a component of the NuA4 HAT complex. These studies suggest that the yeast Ing1 related proteins are involved in chromatin remodeling. They further suggest that these functions may be conserved in mammals and provide a possible mechanism for the human Ing1 candidate tumor suppressor. PMID- 10805725 TI - Class II phosphoinositide 3-kinases are downstream targets of activated polypeptide growth factor receptors. AB - The class II phosphoinositide 3-kinases (PI3K) PI3K-C2alpha and PI3K-C2beta are two recently identified members of the large PI3K family. Both enzymes are characterized by the presence of a C2 domain at the carboxy terminus and, in vitro, preferentially utilize phosphatidylinositol and phosphatidylinositol 4 monophosphate as lipid substrates. Little is understood about how the catalytic activity of either enzyme is regulated in vivo. In this study, we demonstrate that PI3K-C2alpha and PI3K-C2beta represent two downstream targets of the activated epidermal growth factor (EGF) receptor in human carcinoma-derived A431 cells. Stimulation of quiescent cultures with EGF resulted in the rapid recruitment of both enzymes to a phosphotyrosine signaling complex that contained the EGF receptor and Erb-B2. Ligand addition also induced the appearance of a second, more slowly migrating band of PI3K-C2alpha and PI3K-C2beta immunoreactivity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Since both PI3K enzymes can utilize Ca(2+) as an essential divalent cation in lipid kinase assays and since the catalytic activity of PI3K-C2alpha is refractory to the inhibitor wortmannin, these properties were used to confirm the recruitment of each PI3K isozyme to the activated EGF receptor complex. To examine this interaction in greater detail, PI3K-C2beta was chosen for further investigation. EGF and platelet-derived growth factor also stimulated the association of PI3K-C2beta with their respective receptors in other cells, including epithelial cells and fibroblasts. The use of EGF receptor mutants and phosphopeptides derived from the EGF receptor and Erb-B2 demonstrated that the interaction with recombinant PI3K-C2beta occurs through E(p)YL/I phosphotyrosine motifs. The N-terminal region of PI3K-C2beta was found to selectively interact with the EGF receptor in vitro, suggesting that it mediates the association of this PI3K with the receptor. However, the mechanism of this interaction remains unclear. We conclude that class II PI3K enzymes may contribute to the generation of 3' phosphoinositides following the activation of polypeptide growth factor receptors in vivo and thus mediate certain aspects of their biological activity. PMID- 10805726 TI - Two distinct Notch1 mutant alleles are involved in the induction of T-cell leukemia in c-myc transgenic mice. AB - We have previously characterized a large panel of provirus insertion Notch1 mutant alleles and their products arising in thymomas of MMTV(D)/myc transgenic mice. Here, we show that these Notch1 mutations represent two clearly distinct classes. In the first class (type I), proviral integrations were clustered just upstream of sequences encoding the transmembrane domain. Type I Notch1 alleles produced two types of mutant Notch1 RNA, one of which encoded the entire Notch1 cytoplasmic domain [N(IC)] and the other of which encoded a soluble ectodomain [N(EC)(Mut)] which, in contrast to the processed wild-type ectodomain [N(EC)(WT)], did not reside at the cell surface and became secreted in a temperature-dependent manner. A second, novel class of mutant Notch1 allele (type II) encoded a Notch1 receptor with the C-terminal PEST motif deleted (DeltaCT). The type II Notch1(DeltaCT) protein was expressed as a normally processed receptor [N(EC)(WT) and N(IC)(DeltaCT)] at the cell surface, and its ectodomain was found to be shed into the extracellular medium in a temperature- and calcium dependent manner. These data suggest that both type I and type II mutations generate two structurally distinct Notch1 N(EC) and N(IC) proteins that may participate in tumor formation, in collaboration with the c-myc oncogene, through distinct mechanisms. Constitutive type I N(IC) and type II N(IC)(DeltaCT) expression may enhance Notch1 intracellular signaling, while secreted or shed type I N(EC)(Mut) and type II N(EC) proteins may differentially interact in an autocrine or paracrine fashion with ligands of Notch1 and affect their signaling. PMID- 10805727 TI - Repression of ribosome and tRNA synthesis in secretion-defective cells is signaled by a novel branch of the cell integrity pathway. AB - The transcription of ribosomal DNA, ribosomal protein (RP) genes, and 5S and tRNA genes by RNA polymerases (Pols) I, II, and III, respectively, is rapidly and coordinately repressed upon interruption of the secretory pathway in Saccharomyces cerevisiae. We find that repression of ribosome and tRNA synthesis in secretion-defective cells involves activation of the cell integrity pathway. Transcriptional repression requires the upstream components of this pathway, including the Wsc family of putative plasma membrane sensors and protein kinase C (PKC), but not the downstream Bck1-Mkk1/2-Slt2 mitogen-activated protein kinase cascade. These findings reveal a novel PKC effector pathway that controls more than 85% of nuclear transcription. It is proposed that the coordination of ribosome and tRNA synthesis with cell growth may be achieved, in part, by monitoring the turgor pressure of the cell. PMID- 10805728 TI - CD8 coreceptor extinction in signaled CD4(+)CD8(+) thymocytes: coordinate roles for both transcriptional and posttranscriptional regulatory mechanisms in developing thymocytes. AB - T-cell development in the thymus is characterized by changing expression patterns of CD4 and CD8 coreceptor molecules and by changes in CD4 and CD8 gene transcription. In response to T-cell receptor (TCR) signals, thymocytes progress through developmental transitions, such as conversion of CD4(+)CD8(+) (double positive [DP]) thymocytes into intermediate CD4(+)CD8(-) thymocytes, that appear to require more-rapid changes in coreceptor expression than can be accomplished by transcriptional regulation alone. Consequently, we considered the possibility that TCR stimulation of DP thymocytes not only affects coreceptor gene transcription but also affects coreceptor RNA stability. Indeed, we found that TCR signals in DP thymocytes rapidly destabilized preexisting CD4 and CD8 coreceptor RNAs, resulting in their rapid elimination. Destabilization of coreceptor RNA was shown for CD8alpha to be dependent on target sequences in the noncoding region of the RNA. TCR signals also differentially affected coreceptor gene transcription in DP thymocytes, terminating CD8alpha gene transcription but only transiently reducing CD4 gene transcription. Thus, posttranscriptional and transcriptional regulatory mechanisms act coordinately in signaled DP thymocytes to promote the rapid conversion of these cells into intermediate CD4(+)CD8(-) thymocytes. We suggest that destabilization of preexisting coreceptor RNAs is a mechanism by which coreceptor expression in developing thymocytes is rapidly altered at critical points in the differentiation of these cells. PMID- 10805729 TI - DDP1, a heterochromatin-associated multi-KH-domain protein of Drosophila melanogaster, interacts specifically with centromeric satellite DNA sequences. AB - DDP1 is a single-stranded nucleic acid binding protein of Drosophila melanogaster that associates with pericentric heterochromatin. DDP1 contains 15 consecutive KH domains and is homologous to the highly conserved vigilin proteins that, in Saccharomyces cerevisiae, are involved in the control of cell ploidy. DDP1 was identified and purified on the basis of its binding to the pyrimidine-rich C strand of the centromeric Drosophila dodeca-satellite. Here, the interaction of DDP1 with the dodeca-satellite C strand was analyzed in detail. This interaction is sequence specific. In particular, a guanine residue which is highly conserved in natural dodeca-satellite sequences was found to be essential for the efficient binding of DDP1. DDP1 binding was also found to be strongly influenced by the length and extent of secondary structure of the DNA substrate. Efficient DDP1 binding required a minimal length of about 75 to 100 nucleotides and was facilitated by the lack of secondary structure of the substrate. DDP1 also showed a significant affinity for the unstructured pyrimidine-rich strand of the most abundant centromeric Drosophila AAGAG satellite. The stoichiometry of the complexes formed with the dodeca-satellite C strand suggests that, in DDP1, the 15 consecutive KH domains are organized such that they define two nucleic acid binding surfaces. These results are discussed in the context of the possible contribution of DDP1 to heterochromatin organization and function. PMID- 10805730 TI - Molecular determinants of differential ligand sensitivities of insect ecdysteroid receptors. AB - The functional receptor for insect ecdysteroid hormones is a heterodimer consisting of two nuclear hormone receptors, ecdysteroid receptor (EcR) and the retinoid X receptor homologue Ultraspiracle (USP). Although ecdysone is commonly thought to be a hormone precursor and 20-hydroxyecdysone (20E), the physiologically active steroid, little is known about the relative activity of ecdysteroids in various arthropods. As a step toward characterization of potential differential ligand recognition, we have analyzed the activities of various ecdysteroids using gel mobility shift assays and transfection assays in Schneider-2 (S2) cells. Ecdysone showed little activation of the Drosophila melanogaster receptor complex (DmEcR-USP). In contrast, this steroid functioned as a potent ligand for the mosquito Aedes aegypti receptor complex (AaEcR-USP), significantly enhancing DNA binding and transactivating a reporter gene in S2 cells. The mosquito receptor also displayed higher hormone-independent DNA binding activity than the Drosophila receptor. Subunit-swapping experiments indicated that the EcR protein, not the USP protein, was responsible for ligand specificity. Using domain-swapping techniques, we made a series of Aedes and Drosophila EcR chimeric constructs. Differential ligand responsiveness was mapped near the C terminus of the ligand binding domain, within the identity box previously implicated in the dimerization specificity of nuclear receptors. This region includes helices 9 and 10, as determined by comparison with available crystal structures obtained from other nuclear receptors. Site-directed mutagenesis revealed that Phe529 in Aedes EcR, corresponding to Tyr611 in Drosophila EcR, was most critical for ligand specificity and hormone-independent DNA binding activity. These results demonstrated that ecdysone could function as a bona fide ligand in a species-specific manner. PMID- 10805731 TI - Multiple signals regulate GAL transcription in yeast. AB - Gal4p activates transcription of the Saccharomyces GAL genes in response to galactose and is phosphorylated during interaction with the RNA polymerase II (Pol II) holoenzyme. One phosphorylation at S699 is necessary for full GAL induction and is mediated by Srb10p/CDK8 of the RNA Pol II holoenzyme mediator subcomplex. Gal4p S699 phosphorylation is necessary for sensitive response to inducer, and its requirement for GAL induction can be abrogated by high concentrations of galactose in strains expressing wild-type GAL2 and GAL3. Gal4p S699 phosphorylation occurs independently of Gal3p and is responsible for the long-term adaptation response observed in gal3 yeast. SRB10 and GAL3 are shown to represent parallel mechanisms for GAL gene induction. These results demonstrate that Gal4p activity is controlled by two independent signals: one that acts through Gal3p-galactose and a second that is mediated by the holoenzyme associated cyclin-dependent kinase Srb10p. Since Srb10p is regulated independently of galactose, our results suggest a function for CDK8 in coordinating responses to specific inducers with the environment through the phosphorylation of gene-specific activators. PMID- 10805732 TI - Rck2 kinase is a substrate for the osmotic stress-activated mitogen-activated protein kinase Hog1. AB - Exposure of yeast cells to increases in extracellular osmolarity activates the Hog1 mitogen-activated protein kinase (MAPK). Activation of Hog1 MAPK results in induction of a set of osmoadaptive responses, which allow cells to survive in high-osmolarity environments. Little is known about how the MAPK activation results in induction of these responses, mainly because no direct substrates for Hog1 have been reported. We conducted a two-hybrid screening using Hog1 as a bait to identify substrates for the MAPK, and the Rck2 protein kinase was identified as an interactor for Hog1. Both two-hybrid analyses and coprecipitation assays demonstrated that Hog1 binds strongly to the C-terminal region of Rck2. Upon osmotic stress, Rck2 was phosphorylated in vivo in a Hog1-dependent manner. Furthermore, purified Hog1 was able to phosphorylate Rck2 when activated both in vivo and in vitro. Rck2 phosphorylation occurred specifically at Ser519, a residue located within the C-terminal putative autoinhibitory domain. Interestingly, phosphorylation at Ser519 by Hog1 resulted in an increase of Rck2 kinase activity. Overexpression of Rck2 partially suppressed the osmosensitive phenotype of hog1Delta and pbs2Delta cells, suggesting that Rck2 is acting downstream of Hog1. Consistently, growth arrest caused by hyperactivation of the Hog1 MAPK was abolished by deletion of the RCK2 gene. Furthermore, overexpression of a catalytically impaired (presumably dominant inhibitory) Rck2 kinase resulted in a decrease of osmotolerance in wild-type cells but not in hog1Delta cells. Taken together, our data suggest that Rck2 acts downstream of Hog1, controlling a subset of the responses induced by the MAPK upon osmotic stress. PMID- 10805734 TI - Interaction between PAK and nck: a template for Nck targets and role of PAK autophosphorylation. AB - The kinase PAK binds tightly to the SH3 domain of its partner PIX via a central proline-rich sequence. A different N-terminal sequence allows alphaPAK to bind an SH3 domain of the adaptor Nck. The Nck SH3[2] domain interacts equally with an 18 mer PAK-derived peptide and full-length alphaPAK. Detailed analysis of this binding by saturation substitution allows related Nck targets to be accurately identified from sequence characteristics alone. All Nck SH3[2] binding proteins, including PAK, NIK, synaptojanin, PRK2, and WIP, possess the motif PXXPXRXXS; in the case of PAK, serine phosphorylation at this site negatively regulates binding. We show that kinase autophosphorylation blocks binding by both Nck and PIX to alphaPAK, thus providing a mechanism to regulate PAK interactions with its SH3-containing partners. One cellular consequence of the regulatable binding of PAK is facilitation of its cycling between cytosolic and focal complex sites. PMID- 10805733 TI - The differential effects of pp120 (Ceacam 1) on the mitogenic action of insulin and insulin-like growth factor 1 are regulated by the nonconserved tyrosine 1316 in the insulin receptor. AB - pp120 (Ceacam 1) undergoes ligand-stimulated phosphorylation by the insulin receptor, but not by the insulin-like growth factor 1 receptor (IGF-1R). This differential phosphorylation is regulated by the C terminus of the beta-subunit of the insulin receptor, the least conserved domain of the two receptors. In the present studies, deletion and site-directed mutagenesis in stably transfected hepatocytes derived from insulin receptor knockout mice (IR(-/-)) revealed that Tyr(1316), which is replaced by the nonphosphorylatable phenylalanine in IGF-1R, regulated the differential phosphorylation of pp120 by the insulin receptor. Similarly, the nonconserved Tyr(1316) residue also regulated the differential effect of pp120 on IGF-1 and insulin mitogenesis, with pp120 downregulating the growth-promoting action of insulin, but not that of IGF-1. Thus, it appears that pp120 phosphorylation by the insulin receptor is required and sufficient to mediate its downregulatory effect on the mitogenic action of insulin. Furthermore, the current studies revealed that the C terminus of the beta-subunit of the insulin receptor contains elements that suppress the mitogenic action of insulin. Because IR(-/-) hepatocytes are derived from liver, an insulin-targeted tissue, our observations have finally resolved the controversy about the role of the least-conserved domain of insulin and IGF-1Rs in mediating the difference in the mitogenic action of their ligands, with IGF-1 being more mitogenic than insulin. PMID- 10805735 TI - Role of Saccharomyces cerevisiae ISA1 and ISA2 in iron homeostasis. AB - The budding yeast Saccharomyces cerevisiae contains two homologues of bacterial IscA proteins, designated Isa1p and Isa2p. Bacterial IscA is a product of the isc (iron-sulfur cluster) operon and has been suggested to participate in Fe-S cluster formation or repair. To test the function of yeast Isa1p and Isa2p, single or combinatorial disruptions were introduced in ISA1 and ISA2. The resultant isaDelta mutants were viable but exhibited a dependency on lysine and glutamate for growth and a respiratory deficiency due to an accumulation of mutations in mitochondrial DNA. As with other yeast genes proposed to function in Fe-S cluster assembly, mitochondrial iron concentration was significantly elevated in the isa mutants, and the activities of the Fe-S cluster-containing enzymes aconitase and succinate dehydrogenase were dramatically reduced. An inspection of Isa-like proteins from bacteria to mammals revealed three invariant cysteine residues, which in the case of Isa1p and Isa2p are essential for function and may be involved in iron binding. As predicted, Isa1p is targeted to the mitochondrial matrix. However, Isa2p is present within the intermembrane space of the mitochondria. Our deletion analyses revealed that Isa2p harbors a bipartite N-terminal leader sequence containing a mitochondrial import signal linked to a second sequence that targets Isa2p to the intermembrane space. Both signals are needed for Isa2p function. A model for the nonredundant roles of Isa1p and Isa2p in delivering iron to sites of the Fe-S cluster assembly is discussed. PMID- 10805737 TI - Mutational analysis of mammalian translation initiation factor 5 (eIF5): role of interaction between the beta subunit of eIF2 and eIF5 in eIF5 function in vitro and in vivo. AB - Eukaryotic translation initiation factor 5 (eIF5) interacts with the 40S initiation complex (40S-eIF3-AUG-Met-tRNA(f)-eIF2-GTP) to promote the hydrolysis of ribosome-bound GTP. eIF5 also forms a complex with eIF2 by interacting with the beta subunit of eIF2. In this work, we have used a mutational approach to investigate the importance of eIF5-eIF2beta interaction in eIF5 function. Binding analyses with recombinant rat eIF5 deletion mutants identified the C terminus of eIF5 as the eIF2beta-binding region. Alanine substitution mutagenesis at sites within this region defined several conserved glutamic acid residues in a bipartite motif as critical for eIF5 function. The E346A,E347A and E384A,E385A double-point mutations each caused a severe defect in the binding of eIF5 to eIF2beta but not to eIF3-Nip1p, while a eIF5 hexamutant (E345A,E346A, E347A,E384A,E385A,E386A) showed negligible binding to eIF2beta. These mutants were also severely defective in eIF5-dependent GTP hydrolysis, in 80S initiation complex formation, and in the ability to stimulate translation of mRNAs in an eIF5-dependent yeast cell-free translation system. Furthermore, unlike wild-type rat eIF5, which can functionally substitute for yeast eIF5 in complementing in vivo a genetic disruption of the chromosomal copy of the TIF5 gene, the eIF5 double-point mutants allowed only slow growth of this DeltaTIF5 yeast strain, while the eIF5 hexamutant was unable to support cell growth and viability of this strain. These findings suggest that eIF5-eIF2beta interaction plays an essential role in eIF5 function in eukaryotic cells. PMID- 10805736 TI - Neoplastic transformation by Notch requires nuclear localization. AB - Notch proteins are plasma membrane-spanning receptors that mediate important cell fate decisions such as differentiation, proliferation, and apoptosis. The mechanism of Notch signaling remains poorly understood. However, it is clear that the Notch signaling pathway mediates its effects through intercellular contact between neighboring cells. The prevailing model for Notch signaling suggests that ligand, presented on a neighboring cell, triggers proteolytic processing of Notch. Following proteolysis, it is thought that the intracellular portion of Notch (N(ic)) translocates to the nucleus, where it is involved in regulating gene expression. There is considerable debate concerning where in the cell Notch functions and what proteins serve as effectors of the Notch signal. Several Notch genes have clearly been shown to be proto-oncogenes in mammalian cells. Activation of Notch proto-oncogenes has been associated with tumorigenesis in several human and other mammalian cancers. Transforming alleles of Notch direct the expression of truncated proteins that primarily consist of N(ic) and are not tethered to the plasma membrane. However, the mechanism by which Notch oncoproteins (generically termed here as N(ic)) induce neoplastic transformation is not known. Previously we demonstrated that N1(ic) and N2(ic) could transform E1A immortalized baby rat kidney cells (RKE) in vitro. We now report direct evidence that N1(ic) must accumulate in the nucleus to induce transformation of RKE cells. In addition, we define the minimal domain of N1(ic) required to induce transformation and present evidence that transformation of RKE cells by N1(ic) is likely to be through a CBF1-independent pathway. PMID- 10805738 TI - p38 and extracellular signal-regulated kinases regulate the myogenic program at multiple steps. AB - The extracellular signals which regulate the myogenic program are transduced to the nucleus by mitogen-activated protein kinases (MAPKs). We have investigated the role of two MAPKs, p38 and extracellular signal-regulated kinase (ERK), whose activities undergo significant changes during muscle differentiation. p38 is rapidly activated in myocytes induced to differentiate. This activation differs from those triggered by stress and cytokines, because it is not linked to Jun-N terminal kinase stimulation and is maintained during the whole process of myotube formation. Moreover, p38 activation is independent of a parallel promyogenic pathway stimulated by insulin-like growth factor 1. Inhibition of p38 prevents the differentiation program in myogenic cell lines and human primary myocytes. Conversely, deliberate activation of endogenous p38 stimulates muscle differentiation even in the presence of antimyogenic cues. Much evidence indicates that p38 is an activator of MyoD: (i) p38 kinase activity is required for the expression of MyoD-responsive genes, (ii) enforced induction of p38 stimulates the transcriptional activity of a Gal4-MyoD fusion protein and allows efficient activation of chromatin-integrated reporters by MyoD, and (iii) MyoD dependent myogenic conversion is reduced in mouse embryonic fibroblasts derived from p38alpha(-/-) embryos. Activation of p38 also enhances the transcriptional activities of myocyte enhancer binding factor 2A (MEF2A) and MEF2C by direct phosphorylation. With MEF2C, selective phosphorylation of one residue (Thr293) is a tissue-specific activating signal in differentiating myocytes. Finally, ERK shows a biphasic activation profile, with peaks of activity in undifferentiated myoblasts and postmitotic myotubes. Importantly, activation of ERK is inhibitory toward myogenic transcription in myoblasts but contributes to the activation of myogenic transcription and regulates postmitotic responses (i.e., hypertrophic growth) in myotubes. PMID- 10805739 TI - Identification of domains and residues within the epsilon subunit of eukaryotic translation initiation factor 2B (eIF2Bepsilon) required for guanine nucleotide exchange reveals a novel activation function promoted by eIF2B complex formation. AB - Eukaryotic translation initiation factor 2B (eIF2B) is the guanine nucleotide exchange factor for protein synthesis initiation factor 2 (eIF2). Composed of five subunits, it converts eIF2 from a GDP-bound form to the active eIF2-GTP complex. This is a regulatory step of translation initiation. In vitro, eIF2B catalytic function can be provided by the largest (epsilon) subunit alone (eIF2Bepsilon). This activity is stimulated by complex formation with the other eIF2B subunits. We have analyzed the roles of different regions of eIF2Bepsilon in catalysis, in eIF2B complex formation, and in binding to eIF2 by characterizing mutations in the Saccharomyces cerevisiae gene encoding eIF2Bepsilon (GCD6) that impair the essential function of eIF2B. Our analysis of nonsense mutations indicates that the C terminus of eIF2Bepsilon (residues 518 to 712) is required for both catalytic activity and interaction with eIF2. In addition, missense mutations within this region impair the catalytic activity of eIF2Bepsilon without affecting its ability to bind eIF2. Internal, in-frame deletions within the N-terminal half of eIF2Bepsilon disrupt eIF2B complex formation without affecting the nucleotide exchange activity of eIF2Bepsilon alone. Finally, missense mutations identified within this region do not affect the catalytic activity of eIF2Bepsilon alone or its interactions with the other eIF2B subunits or with eIF2. Instead, these missense mutations act indirectly by impairing the enhancement of the rate of nucleotide exchange that results from complex formation between eIF2Bepsilon and the other eIF2B subunits. This suggests that the N-terminal region of eIF2Bepsilon is an activation domain that responds to eIF2B complex formation. PMID- 10805740 TI - Conversion of topoisomerase I cleavage complexes on the leading strand of ribosomal DNA into 5'-phosphorylated DNA double-strand breaks by replication runoff. AB - Topoisomerase I cleavage complexes can be induced by a variety of DNA damages and by the anticancer drug camptothecin. We have developed a ligation-mediated PCR (LM-PCR) assay to analyze replication-mediated DNA double-strand breaks induced by topoisomerase I cleavage complexes in human colon carcinoma HT29 cells at the nucleotide level. We found that conversion of topoisomerase I cleavage complexes into replication-mediated DNA double-strand breaks was only detectable on the leading strand for DNA synthesis, which suggests an asymmetry in the way that topoisomerase I cleavage complexes are metabolized on the two arms of a replication fork. Extension by Taq DNA polymerase was not required for ligation to the LM-PCR primer, indicating that the 3' DNA ends are extended by DNA polymerase in vivo closely to the 5' ends of the topoisomerase I cleavage complexes. These findings suggest that the replication-mediated DNA double-strand breaks generated at topoisomerase I cleavage sites are produced by replication runoff. We also found that the 5' ends of these DNA double-strand breaks are phosphorylated in vivo, which suggests that a DNA 5' kinase activity acts on the double-strand ends generated by replication runoff. The replication-mediated DNA double-strand breaks were rapidly reversible after cessation of the topoisomerase I cleavage complexes, suggesting the existence of efficient repair pathways for removal of topoisomerase I-DNA covalent adducts in ribosomal DNA. PMID- 10805741 TI - A 5' splice site-proximal enhancer binds SF1 and activates exon bridging of a microexon. AB - Internal exon size in vertebrates occurs over a narrow size range. Experimentally, exons shorter than 50 nucleotides are poorly included in mRNA unless accompanied by strengthened splice sites or accessory sequences that act as splicing enhancers, suggesting steric interference between snRNPs and other splicing factors binding simultaneously to the 3' and 5' splice sites of microexons. Despite these problems, very small naturally occurring exons exist. Here we studied the factors and mechanism involved in recognizing a constitutively included six-nucleotide exon from the cardiac troponin T gene. Inclusion of this exon is dependent on an enhancer located downstream of the 5' splice site. This enhancer contains six copies of the simple sequence GGGGCUG. The enhancer activates heterologous microexons and will work when located either upstream or downstream of the target exon, suggesting an ability to bind factors that bridge splicing units. A single copy of this sequence is sufficient for in vivo exon inclusion and is the binding site for the known bridging mammalian splicing factor 1 (SF1). The enhancer and its bound SF1 act to increase recognition of the upstream exon during exon definition, such that competition of in vitro reactions with RNAs containing the GGGGCUG repeated sequence depress splicing of the upstream intron, assembly of the spliceosome on the 3' splice site of the exon, and cross-linking of SF1. These results suggest a model in which SF1 bridges the small exon during initial assembly, thereby effectively extending the domain of the exon. PMID- 10805742 TI - Nuclear export of heat shock and non-heat-shock mRNA occurs via similar pathways. AB - Several studies of the yeast Saccharomyces cerevisiae support differential regulation of heat shock mRNA (hs mRNA) and non-hs mRNA nuclear export during stress. These include the finding that hs mRNA export at 42 degrees C is inhibited in the absence of the nucleoporinlike protein Rip1p (also called Nup42p) (C. A. Saavedra, C. M. Hammell, C. V. Heath, and C. N. Cole, Genes Dev. 11:2845-2856, 1997; F. Stutz, J. Kantor, D. Zhang, T. McCarthy, M. Neville, and M. Rosbash, Genes Dev. 11:2857-2868, 1997). However, the results reported in this paper provide little evidence for selective non-hs mRNA retention or selective hs mRNA export under heat shock conditions. First, we do not detect a block to non hs mRNA export at 42 degrees C in a wild-type strain. Second, hs mRNA export appears to be mediated by the Ran system and several other factors previously reported to be important for general mRNA export. Third, the export of non-hs mRNA as well as hs mRNA is inhibited in the absence of Rip1p at 42 degrees C. As a corollary, we find no evidence for cis-acting hs mRNA sequences that promote transport during heat shock. Taken together, our data suggest that a shift to 42 degrees C in the absence of Rip1p impacts a late stage of transport affecting most if not all mRNA. PMID- 10805743 TI - Saccharomyces cerevisiae RAI1 (YGL246c) is homologous to human DOM3Z and encodes a protein that binds the nuclear exoribonuclease Rat1p. AB - The RAT1 gene of Saccharomyces cerevisiae encodes a 5'-->3' exoribonuclease which plays an essential role in yeast RNA degradation and/or processing in the nucleus. We have cloned a previously uncharacterized gene (YGL246c) that we refer to as RAI1 (Rat1p interacting protein 1). RAI1 is homologous to Caenorhabditis elegans DOM-3 and human DOM3Z. Deletion of RAI1 confers a growth defect which can be complemented by an additional copy of RAT1 on a centromeric vector or by directing Xrn1p, the cytoplasmic homolog of Rat1p, to the nucleus through the addition of a nuclear targeting sequence. Deletion of RAI1 is synthetically lethal with the rat1-1(ts) mutation and shows genetic interaction with a deletion of SKI2 but not XRN1. Polysome analysis of an rai1 deletion mutant indicated a defect in 60S biogenesis which was nearly fully reversed by high-copy RAT1. Northern blot analysis of rRNAs revealed that rai1 is required for normal 5.8S processing. In the absence of RAI1, 5.8S(L) was the predominant form of 5.8S and there was an accumulation of 3'-extended forms but not 5'-extended species of 5. 8S. In addition, a 27S pre-rRNA species accumulated in the rai1 mutant. Thus, deletion of RAI1 affects both 5' and 3' processing reactions of 5.8S rRNA. Consistent with the in vivo data suggesting that RAI1 enhances RAT1 function, purified Rai1p stabilized the in vitro exoribonuclease activity of Rat1p. PMID- 10805744 TI - Cpc2, a fission yeast homologue of mammalian RACK1 protein, interacts with Ran1 (Pat1) kinase To regulate cell cycle progression and meiotic development. AB - The Schizosaccharomyces pombe ran1/pat1 gene regulates the transition between mitosis and meiosis. Inactivation of Ran1 (Pat1) kinase is necessary and sufficient for cells to exit the cell cycle and undergo meiosis. The yeast two hybrid interaction trap was used to identify protein partners for Ran1/Pat1. Here we report the identification of one of these, Cpc2. Cpc2 encodes a homologue of RACK1, a WD protein with homology to the beta subunit of heterotrimeric G proteins. RACK1 is a highly conserved protein, although its function remains undefined. In mammalian cells, RACK1 physically associates with some signal transduction proteins, including Src and protein kinase C. Fission yeast cells containing a cpc2 null allele are viable but cell cycle delayed. cpc2Delta cells fail to accumulate in G(1) when starved of nitrogen. This leads to defects in conjugation and meiosis. Copurification studies show that although Cpc2 and Ran1 (Pat1) physically associate, Cpc2 does not alter Ran1 (Pat1) kinase activity in vitro. Using a Ran1 (Pat1) fusion to green fluorescent protein, we show that localization of the kinase is impaired in cpc2Delta cells. Thus, in parallel with the proposed role of RACK1 in mammalian cells, fission yeast cpc2 may function as an anchoring protein for Ran1 (Pat1) kinase. All defects associated with loss of cpc2 are reversed in cells expressing mammalian RACK1, demonstrating that the fission yeast and mammalian gene products are indeed functional homologues. PMID- 10805745 TI - Homologous and nonhomologous recombination resulting in deletion: effects of p53 status, microhomology, and repetitive DNA length and orientation. AB - Repetitive DNA elements frequently are precursors to chromosomal deletions in prokaryotes and lower eukaryotes. However, little is known about the relationship between repeated sequences and deletion formation in mammalian cells. We have created a novel integrated plasmid-based recombination assay to investigate repeated sequence instability in human cells. In a control cell line, the presence of direct or inverted repeats did not appreciably influence the very low deletion frequencies (2 x 10(-7) to 9 x 10(-7)) in the region containing the repeat. Similar to what has been observed in lower eukaryotes, the majority of deletions resulted from the loss of the largest direct repeat present in the system along with the intervening sequence. Interestingly, in closely related cell lines that possess a mutant p53 gene, deletion frequencies in the control and direct-repeat plasmids were 40 to 300 times higher than in their wild-type counterparts. However, mutant p53 cells did not preferentially utilize the largest available homology in the formation of the deletion. Surprisingly, inverted repeats were approximately 10,000 times more unstable in all mutant p53 cells than in wild-type cells. Finally, several deletion junctions were marked by the addition of novel bases that were homologous to one of the preexisting DNA ends. Contrary to our expectations, only 6% of deletions in all cell lines could be classified as arising from nonhomologous recombination. PMID- 10805746 TI - Aberrant splicing of tau pre-mRNA caused by intronic mutations associated with the inherited dementia frontotemporal dementia with parkinsonism linked to chromosome 17. AB - Frontotemporal dementia accounts for a significant fraction of dementia cases. Frontotemporal dementia with parkinsonism linked to chromosome 17 is associated with either exonic or intronic mutations in the tau gene. This highlights the involvement of aberrant pre-mRNA splicing in the pathogenesis of neurodegenerative disorders. Little is known about the molecular mechanisms of the splicing defects underlying these diseases. To establish a model system for studying the role of pre-mRNA splicing in neurodegenerative diseases, we have constructed a tau minigene that reproduces tau alternative splicing in both cultured cells and in vitro biochemical assays. We demonstrate that mutations in a nonconserved intronic region of the human tau gene lead to increased splicing between exon 10 and exon 11. Systematic biochemical analyses indicate the importance of U1 snRNP and, to a lesser extent, U6 snRNP in differentially recognizing wild-type versus intron mutant tau pre-mRNAs. Gel mobility shift assays with purified U1 snRNP and oligonucleotide-directed RNase H cleavage experiments support the idea that the intronic mutations destabilize a stem-loop structure that sequesters the 5' splice site downstream of exon 10 in tau pre mRNA, leading to increases in U1 snRNP binding and in splicing between exon 10 and exon 11. Thus, mutations in nonconserved intronic regions that increase rather than decrease alternative splicing can be an important pathogenic mechanism for the development of human diseases. PMID- 10805747 TI - Septin-dependent assembly of a cell cycle-regulatory module in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae septin mutants have pleiotropic defects, which include the formation of abnormally elongated buds. This bud morphology results at least in part from a cell cycle delay imposed by the Cdc28p-inhibitory kinase Swe1p. Mutations in three other genes (GIN4, encoding a kinase related to the Schizosaccharomyces pombe mitotic inducer Nim1p; CLA4, encoding a p21-activated kinase; and NAP1, encoding a Clb2p-interacting protein) also produce perturbations of septin organization associated with an Swe1p-dependent cell cycle delay. The effects of gin4, cla4, and nap1 mutations are additive, indicating that these proteins promote normal septin organization through pathways that are at least partially independent. In contrast, mutations affecting the other two Nim1p-related kinases in S. cerevisiae, Hsl1p and Kcc4p, produce no detectable effect on septin organization. However, deletion of HSL1, but not of KCC4, did produce a cell cycle delay under some conditions; this delay appears to reflect a direct role of Hsl1p in the regulation of Swe1p. As shown previously, Swe1p plays a central role in the morphogenesis checkpoint that delays the cell cycle in response to defects in bud formation. Swe1p is localized to the nucleus and to the daughter side of the mother bud neck prior to its degradation in G(2)/M phase. Both the neck localization of Swe1p and its degradation require Hsl1p and its binding partner Hsl7p, both of which colocalize with Swe1p at the daughter side of the neck. This localization is lost in mutants with perturbed septin organization, suggesting that the release of Hsl1p and Hsl7p from the neck may reduce their ability to inactivate Swe1p and thus contribute to the G(2) delay observed in such mutants. In contrast, treatments that perturb actin organization have little effect on Hsl1p and Hsl7p localization, suggesting that such treatments must stabilize Swe1p by another mechanism. The apparent dependence of Swe1p degradation on localization of the Hsl1p-Hsl7p-Swe1p module to a site that exists only in budded cells may constitute a mechanism for deactivating the morphogenesis checkpoint when it is no longer needed (i.e., after a bud has formed). PMID- 10805748 TI - Interference of the simian virus 40 origin of replication by the cytomegalovirus immediate early gene enhancer: evidence for competition of active regulatory chromatin conformation in a single domain. AB - Replication origins are often found closely associated with transcription regulatory elements in both prokaryotic and eukaryotic cells. To examine the relationship between these two elements, we studied the effect of a strong promoter-enhancer on simian virus 40 (SV40) DNA replication. The human cytomegalovirus (CMV) immediate early gene enhancer-promoter was found to exert a strong inhibitory effect on SV40 origin-based plasmid replication in Cos-1 cells in a position- and dose-dependent manner. Deletion analysis indicated that the effect was exerted by sequences located in the enhancer portion of the CMV sequence, thus excluding the mechanism of origin occlusion by transcription. Insertion of extra copies of the SV40 origin only partially alleviated the inhibition. Analysis of nuclease-sensitive cleavage sites of chromatin containing the transfected plasmids indicate that the chromatin was cleaved at one of the regulatory sites in the plasmids containing more than one regulatory site, suggesting that only one nuclease-hypersensitive site existed per chromatin. A positive correlation was found between the degree of inhibition of DNA replication and the decrease of P1 cleavage frequency at the SV40 origin. The CMV enhancer was also found to exhibit an inhibitory effect on the CMV enhancer promoter driving chloramphenicol acetyltransferase expression in a dose-dependent manner. Together these results suggest that inhibition of SV40 origin-based DNA replication by the CMV enhancer is due to intramolecular competition for the formation of active chromatin structure. PMID- 10805749 TI - The p53 response to DNA damage in vivo is independent of DNA-dependent protein kinase. AB - Ionizing radiation (IR) exposure causes mammalian cells to undergo p53-dependent cell cycle arrest and/or apoptosis. The in vivo role of DNA-dependent protein kinase (DNA-PK) in the transduction of the DNA damage signal to p53 remains unresolved. To determine the relationship between DNA-PK and p53, we studied the cell cycle and apoptotic responses to IR in mice deficient in DNA-PK. Using the slip mouse, which harbors an inactivating mutation of the DNA-PK catalytic subunit (DNA-PKcs), we demonstrated not only that these DNA-PKcs null mutants were highly radiosensitive but also that upon IR treatment, p53 accumulated in their cultured cells and tissue. Induced p53 was transcriptionally active and mediated the induction of p21 and Bax in slip cells. Examination of the thymic cell cycle response to IR treatment indicated that the slip G(1)/S-phase cell cycle checkpoint function was intact. We further show that slip mice exhibited a higher level of spontaneous thymic apoptosis as well as a more robust apoptotic response to IR than wild-type mice. Together, these data demonstrate that the p53 mediated response to DNA damage is intact in cells devoid of DNA-PK activity and suggest that other kinases, such as the product of the gene (ATM) mutated in ataxia telangiectasia, are better candidates for regulating IR-induced phosphorylation and accumulation of p53. PMID- 10805750 TI - Prolyl 4-hydroxylase is an essential procollagen-modifying enzyme required for exoskeleton formation and the maintenance of body shape in the nematode Caenorhabditis elegans. AB - The multienzyme complex prolyl 4-hydroxylase catalyzes the hydroxylation of proline residues and acts as a chaperone during collagen synthesis in multicellular organisms. The beta subunit of this complex is identical to protein disulfide isomerase (PDI). The free-living nematode Caenorhabditis elegans is encased in a collagenous exoskeleton and represents an excellent model for the study of collagen biosynthesis and extracellular matrix formation. In this study, we examined prolyl 4-hydroxylase alpha-subunit (PHY; EC 1.14.11.2)- and beta subunit (PDI; EC 5.3.4.1)-encoding genes with respect to their role in collagen modification and formation of the C. elegans exoskeleton. We identified genes encoding two PHYs and a single associated PDI and showed that all three are expressed in collagen-synthesizing ectodermal cells at times of maximal collagen synthesis. Disruption of the pdi gene via RNA interference resulted in embryonic lethality. Similarly, the combined phy genes are required for embryonic development. Interference with phy-1 resulted in a morphologically dumpy phenotype, which we determined to be identical to the uncharacterized dpy-18 locus. Two dpy-18 mutant strains were shown to have null alleles for phy-1 and to have a reduced hydroxyproline content in their exoskeleton collagens. This study demonstrates in vivo that this enzyme complex plays a central role in extracellular matrix formation and is essential for normal metazoan development. PMID- 10805751 TI - hnRNP C is required for postimplantation mouse development but Is dispensable for cell viability. AB - The hnRNP C1 and C2 proteins are among the most abundant proteins in the nucleus, and as ubiquitous components of RNP complexes, they have been implicated in many aspects of mRNA biogenesis. In this report, we have characterized a null mutation induced in embryonic stem cells by insertion of the U3His gene trap retrovirus into the first intron of the hnRNP C1/C2 gene. cDNAs encoding murine hnRNP C1 and C2 were characterized, and the predicted protein sequences were found to be highly conserved among vertebrates. A human consensus sequence, generated from over 400 expressed sequence tags, suggests two revisions to the previously published human sequence. In addition, alternatively spliced transcripts, expressed only by the murine gene, encode four novel proteins: variants of C1 and C2 with either seven additional amino acids or one fewer amino acid in a region between the oligomerization and C-terminal acidic domains. The disrupted gene was transmitted into the germ line and is tightly linked to a recessive, embryonic lethal phenotype. Homozygous mutant embryos fail to develop beyond the egg cylinder stage and are resorbed by 10.5 days of gestation, a phenotype consistent with a fundamental role in cellular metabolism. However, hnRNP C1 and C2 are not required for cell viability. Embryonic stem cell lines established from homozygous mutant blastocysts did not express detectable levels of either protein yet were able to grow and differentiate in vitro, albeit more slowly than wild type cells. These results indicate that the C1 and C2 hnRNPs are not required for any essential step in mRNA biogenesis; however, the proteins may influence the rate and/or fidelity of one or more steps. PMID- 10805752 TI - Analysis of fractalkine receptor CX(3)CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. AB - The seven-transmembrane receptor CX(3)CR1 is a specific receptor for the novel CX(3)C chemokine fractalkine (FKN) (neurotactin). In vitro data suggest that membrane anchoring of FKN, and the existence of a shed, soluble FKN isoform allow for both adhesive and chemoattractive properties. Expression on activated endothelium and neurons defines FKN as a potential target for therapeutic intervention in inflammatory conditions, particularly central nervous system diseases. To investigate the physiological function of CX(3)CR1-FKN interactions, we generated a mouse strain in which the CX(3)CR1 gene was replaced by a green fluorescent protein (GFP) reporter gene. In addition to the creation of a mutant CX(3)CR1 locus, this approach enabled us to assign murine CX(3)CR1 expression to monocytes, subsets of NK and dendritic cells, and the brain microglia. Analysis of CX(3)CR1-deficient mice indicates that CX(3)CR1 is the only murine FKN receptor. Yet, defying anticipated FKN functions, absence of CX(3)CR1 interferes neither with monocyte extravasation in a peritonitis model nor with DC migration and differentiation in response to microbial antigens or contact sensitizers. Furthermore, a prominent response of CX(3)CR1-deficient microglia to peripheral nerve injury indicates unimpaired neuronal-glial cross talk in the absence of CX(3)CR1. PMID- 10805753 TI - Telomere maintenance in telomerase-deficient mouse embryonic stem cells: characterization of an amplified telomeric DNA. AB - Telomere dynamics, chromosomal instability, and cellular viability were studied in serial passages of mouse embryonic stem (ES) cells in which the telomerase RNA (mTER) gene was deleted. These cells lack detectable telomerase activity, and their growth rate was reduced after more than 300 divisions and almost zero after 450 cell divisions. After this growth crisis, survivor cells with a rapid growth rate did emerge. Such survivors were found to maintain functional telomeres in a telomerase-independent fashion. Although telomerase-independent telomere maintenance has been reported for some immortalized mammalian cells, its molecular mechanism has not been elucidated. Characterization of the telomeric structures in one of the survivor mTER(-/-) cell lines showed amplification of the same tandem arrays of telomeric and nontelomeric sequences at most of the chromosome ends. This evidence implicates cis/trans amplification as one mechanism for the telomerase-independent maintenance of telomeres in mammalian cells. PMID- 10805754 TI - A novel chromodomain protein, pdd3p, associates with internal eliminated sequences during macronuclear development in Tetrahymena thermophila. AB - Conversion of the germ line micronuclear genome into the genome of a somatic macronucleus in Tetrahymena thermophila requires several DNA rearrangement processes. These include (i) excision and subsequent elimination of several thousand internal eliminated sequences (IESs) scattered throughout the micronuclear genome and (ii) breakage of the micronuclear chromosomes into hundreds of DNA fragments, followed by de novo telomere addition to their ends. Chromosome breakage sequences (Cbs) that determine the sites of breakage and short regions of DNA adjacent to them are also eliminated. Both processes occur concomitantly in the developing macronucleus. Two stage-specific protein factors involved in germ line DNA elimination have been described previously. Pdd1p and Pdd2p (for programmed DNA degradation) physically associate with internal eliminated sequences in transient electron-dense structures in the developing macronucleus. Here, we report the purification, sequence analysis, and characterization of Pdd3p, a novel developmentally regulated, chromodomain containing polypeptide. Pdd3p colocalizes with Pdd1p in the peripheral regions of DNA elimination structures, but is also found more internally. DNA cross-linked and immunoprecipitated with Pdd1p- or Pdd3p-specific antibodies is enriched in IESs, but not Cbs, suggesting that different protein factors are involved in elimination of these two groups of sequences. PMID- 10805755 TI - Altered activity, social behavior, and spatial memory in mice lacking the NTAN1p amidase and the asparagine branch of the N-end rule pathway. AB - The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. N-terminal asparagine and glutamine are tertiary destabilizing residues, in that they are enzymatically deamidated to yield secondary destabilizing residues aspartate and glutamate, which are conjugated to arginine, a primary destabilizing residue. N-terminal arginine of a substrate protein is bound by the Ubr1-encoded E3alpha, the E3 component of the ubiquitin proteasome-dependent N-end rule pathway. We describe the construction and analysis of mouse strains lacking the asparagine-specific N-terminal amidase (Nt(N)-amidase), encoded by the Ntan1 gene. In wild-type embryos, Ntan1 was strongly expressed in the branchial arches and in the tail and limb buds. The Ntan1(-/-) mouse strains lacked the Nt(N)-amidase activity but retained glutamine specific Nt(Q)-amidase, indicating that the two enzymes are encoded by different genes. Among the normally short-lived N-end rule substrates, only those bearing N terminal asparagine became long-lived in Ntan1(-/-) fibroblasts. The Ntan1(-/-) mice were fertile and outwardly normal but differed from their congenic wild-type counterparts in spontaneous activity, spatial memory, and a socially conditioned exploratory phenotype that has not been previously described with other mouse strains. PMID- 10805756 TI - Phosphorylation of SOX9 by cyclic AMP-dependent protein kinase A enhances SOX9's ability to transactivate a Col2a1 chondrocyte-specific enhancer. AB - Sox9 is a high-mobility-group domain-containing transcription factor required for chondrocyte differentiation and cartilage formation. We used a yeast two-hybrid method based on Son of Sevenless (SOS) recruitment to screen a chondrocyte cDNA library and found that the catalytic subunit of cyclic AMP (cAMP)-dependent protein kinase A (PKA-Calpha) interacted specifically with SOX9. Next we found that two consensus PKA phosphorylation sites within SOX9 could be phosphorylated by PKA in vitro and that SOX9 could be phosphorylated by PKA-Calpha in vivo. In COS-7 cells cotransfected with PKA-Calpha and SOX9 expression plasmids, PKA enhanced the phosphorylation of wild-type SOX9 but did not affect phosphorylation of a SOX9 protein in which the two PKA phosphorylation sites (S(64) and S(211)) were mutated. Using a phosphospecific antibody that specifically recognized SOX9 phosphorylated at serine 211, one of the two PKA phosphorylation sites, we demonstrated that addition of cAMP to chondrocytes strongly increased the phosphorylation of endogenous Sox9. In addition, immunohistochemistry of mouse embryo hind legs showed that Sox9 phosphorylated at serine 211 was principally localized in the prehypertrophic zone of the growth plate, corresponding to the major site of expression of the parathyroid hormone-related peptide (PTHrP) receptor. Since cAMP has previously been shown to effectively increase the mRNA levels of Col2a1 and other specific markers of chondrocyte differentiation in culture, we then asked whether PKA phosphorylation could modulate the activity of SOX9. Addition of 8-bromo-cAMP to chondrocytes in culture increased the activity of a transiently transfected SOX9-dependent 48-bp Col2a1 chondrocyte-specific enhancer; similarly, cotransfection of PKA-Calpha increased the activity of this enhancer. Mutations of the two PKA phosphorylation consensus sites of SOX9 markedly decreased the PKA-Calpha activation of this enhancer by SOX9. PKA phosphorylation and the mutations in the consensus PKA phosphorylation sites of SOX9 did not alter its nuclear localization. In vitro phosphorylation of SOX9 by PKA resulted in more efficient DNA binding. We conclude that SOX9 is a target of cAMP signaling and that phosphorylation of SOX9 by PKA enhances its transcriptional and DNA-binding activity. Because PTHrP signaling is mediated by cAMP, our results support the hypothesis that Sox9 is a target of PTHrP signaling in the growth plate and that the increased activity of Sox9 might mediate the effect of PTHrP in maintaining the cells as nonhypertrophic chondrocytes. PMID- 10805757 TI - Regulated nuclear-cytoplasmic localization of interferon regulatory factor 3, a subunit of double-stranded RNA-activated factor 1. AB - Viral double-stranded RNA (dsRNA) generated during the course of infection leads to the activation of a latent transcription factor, dsRNA-activated factor 1 (DRAF1). DRAF1 binds to a DNA target containing the type I interferon-stimulated response element and induces transcription of responsive genes. DRAF1 is a multimeric transcription factor containing the interferon regulatory factor 3 (IRF-3) protein and one of the histone acetyl transferases, CREB binding protein (CBP) or p300 (CBP/p300). In uninfected cells, the IRF-3 component of DRAF1 resides in the cytoplasm. The cytoplasmic localization of IRF-3 is dependent on a nuclear export signal, and we demonstrate IRF-3 recognition by the chromosome region maintenance 1 (CRM1) (also known as exportin 1) shuttling receptor. Following infection and specific phosphorylation, IRF-3 accumulates in the nucleus where it associates with CBP and p300. We identify a nuclear localization signal (NLS) in IRF-3 that is critical for nuclear accumulation. Mutation of the NLS abrogates nuclear localization even following infection. The NLS appears to be active constitutively, but it is recognized by only a subset of importin-alpha shuttling receptors. Evidence is presented to support a model in which IRF-3 normally shuttles between the nucleus and the cytoplasm but cytoplasmic localization is dominant prior to infection. Following infection, phosphorylated IRF-3 can bind to the CBP/p300 proteins resident in the nucleus. We provide the evidence of a role for CBP/p300 binding in the nuclear sequestration of a transcription factor that normally resides in the cytoplasm. PMID- 10805758 TI - S-Phase progression mediates activation of a silenced gene in synthetic nuclei. AB - Aberrant expression of developmentally silenced genes, characteristic of tumor cells and regenerating tissue, is highly correlated with increased cell proliferation. By modeling this process in vitro in synthetic nuclei, we find that DNA replication leads to deregulation of established developmental expression patterns. Chromatin assembly in the presence of adult mouse liver nuclear extract mediates developmental stage-specific silencing of the tumor marker gene alpha-fetoprotein (AFP). Replication of silenced AFP chromatin in synthetic nuclei depletes sequence-specific transcription repressors, thereby disrupting developmentally regulated repression. Hepatoma-derived factors can target partial derepression of AFP, but full transcription activation requires DNA replication. Thus, unscheduled entry into S phase directly mediates activation of a developmentally silenced gene by (i) depleting developmental stage-specific transcription repressors and (ii) facilitating binding of transactivators. PMID- 10805759 TI - Sortase, a universal target for therapeutic agents against gram-positive bacteria? PMID- 10805760 TI - Supramolecular complexes in photosynthetic bacteria. PMID- 10805761 TI - Microcolumns in the cerebral cortex. PMID- 10805762 TI - Gene therapy for the hemoglobin disorders: past, present, and future. PMID- 10805763 TI - The making of infectious viral RNA: No size limit in sight. PMID- 10805764 TI - Correlation between the structure of the bacterial peptidoglycan monomer unit, the specificity of transpeptidation, and susceptibility to beta-lactams. PMID- 10805765 TI - Cell signaling pathways as control modules: complexity for simplicity? PMID- 10805766 TI - Description of microcolumnar ensembles in association cortex and their disruption in Alzheimer and Lewy body dementias. AB - The cortex of the brain is organized into clear horizontal layers, laminae, which subserve much of the connectional anatomy of the brain. We hypothesize that there is also a vertical anatomical organization that might subserve local interactions of neuronal functional units, in accord with longstanding electrophysiological observations. We develop and apply a general quantitative method, inspired by analogous methods in condensed matter physics, to examine the anatomical organization of the cortex in human brain. We find, in addition to obvious laminae, anatomical evidence for tightly packed microcolumnar ensembles containing approximately 11 neurons, with a periodicity of about 80 microm. We examine the structural integrity of this new architectural feature in two common dementing illnesses, Alzheimer disease and dementia with Lewy bodies. In Alzheimer disease, there is a dramatic, nearly complete loss of microcolumnar ensemble organization. The relative degree of loss of microcolumnar ensembles is directly proportional to the number of neurofibrillary tangles, but not related to the amount of amyloid-beta deposition. In dementia with Lewy bodies, a similar disruption of microcolumnar ensemble architecture occurs despite minimal neuronal loss. These observations show that quantitative analysis of complex cortical architecture can be applied to analyze the anatomical basis of brain disorders. PMID- 10805767 TI - Exceptionally potent inhibitors of fatty acid amide hydrolase: the enzyme responsible for degradation of endogenous oleamide and anandamide. AB - The development of exceptionally potent inhibitors of fatty acid amide hydrolase (FAAH), the enzyme responsible for the degradation of oleamide (an endogenous sleep-inducing lipid), and anandamide (an endogenous ligand for cannabinoid receptors) is detailed. The inhibitors may serve as useful tools to clarify the role of endogenous oleamide and anandamide and may prove to be useful therapeutic agents for the treatment of sleep disorders or pain. The combination of several features-an optimal C12-C8 chain length, pi-unsaturation introduction at the corresponding arachidonoyl Delta(8,9)/Delta(11,12) and oleoyl Delta(9,10) location, and an alpha-keto N4 oxazolopyridine with incorporation of a second weakly basic nitrogen provided FAAH inhibitors with K(i)s that drop below 200 pM and are 10(2)-10(3) times more potent than the corresponding trifluoromethyl ketones. PMID- 10805769 TI - New patterns of centrifugally driven thermal convection. AB - An experimental study is described of convection driven by thermal buoyancy in the annular gap between two corotating coaxial cylinders, heated from the outside and cooled from the inside. Steady convection patterns of the hexaroll and of the knot type are observed in the case of high Prandtl number fluids, for which the Coriolis force is sufficiently small. Oblique rolls and phase turbulence in the form of irregular patterns of convection can also be observed in wide regions of the parameter space. PMID- 10805768 TI - Selective disruption of protein aggregation by cyclodextrin dimers. AB - Beta-cyclodextrin (CD) dimers (n = 11) were synthesized and tested against eight enzymes, seven of which were dimeric or tetrameric, for inhibitor activity. Initial screening showed that only L-lactate dehydrogenase and citrate synthase were inhibited but only by two specific CD dimers in which two beta-CDs were linked on the secondary face by a pyridine-2,6-dicarboxylic group. Further investigation suggested that these CD dimers inhibit the activity of L-lactate dehydrogenase and citrate synthase at least in part by disruption of protein protein aggregation. PMID- 10805770 TI - Binding of bisubstrate analog promotes large structural changes in the unregulated catalytic trimer of aspartate transcarbamoylase: implications for allosteric regulation. AB - A central problem in understanding enzyme regulation is to define the conformational states that account for allosteric changes in catalytic activity. For Escherichia coli aspartate transcarbamoylase (ATCase; EC) the active, relaxed (R state) holoenzyme is generally assumed to be represented by the crystal structure of the complex of the holoenzyme with the bisubstrate analog N phosphonacetyl-L-aspartate (PALA). It is unclear, however, which conformational differences between the unliganded, inactive, taut (T state) holoenzyme and the PALA complex are attributable to localized effects of inhibitor binding as contrasted to the allosteric transition. To define the conformational changes in the isolated, nonallosteric C trimer resulting from the binding of PALA, we determined the 1.95-A resolution crystal structure of the C trimer-PALA complex. In contrast to the free C trimer, the PALA-bound trimer exhibits approximate threefold symmetry. Conformational changes in the C trimer upon PALA binding include ordering of two active site loops and closure of the hinge relating the N and C-terminal domains. The C trimer-PALA structure closely resembles the liganded C subunits in the PALA-bound holoenzyme. This similarity suggests that the pronounced hinge closure and other changes promoted by PALA binding to the holoenzyme are stabilized by ligand binding. Consequently, the conformational changes attributable to the allosteric transition of the holoenzyme remain to be defined. PMID- 10805771 TI - Uracil-DNA glycosylase-DNA substrate and product structures: conformational strain promotes catalytic efficiency by coupled stereoelectronic effects. AB - Enzymatic transformations of macromolecular substrates such as DNA repair enzyme/DNA transformations are commonly interpreted primarily by active-site functional-group chemistry that ignores their extensive interfaces. Yet human uracil-DNA glycosylase (UDG), an archetypical enzyme that initiates DNA base excision repair, efficiently excises the damaged base uracil resulting from cytosine deamination even when active-site functional groups are deleted by mutagenesis. The 1.8-A resolution substrate analogue and 2.0-A resolution cleaved product cocrystal structures of UDG bound to double-stranded DNA suggest enzyme DNA substrate-binding energy from the macromolecular interface is funneled into catalytic power at the active site. The architecturally stabilized closing of UDG enforces distortions of the uracil and deoxyribose in the flipped-out nucleotide substrate that are relieved by glycosylic bond cleavage in the product complex. This experimentally defined substrate stereochemistry implies the enzyme alters the orientation of three orthogonal electron orbitals to favor electron transpositions for glycosylic bond cleavage. By revealing the coupling of this anomeric effect to a delocalization of the glycosylic bond electrons into the uracil aromatic system, this structurally implicated mechanism resolves apparent paradoxes concerning the transpositions of electrons among orthogonal orbitals and the retention of catalytic efficiency despite mutational removal of active site functional groups. These UDG/DNA structures and their implied dissociative excision chemistry suggest biology favors a chemistry for base-excision repair initiation that optimizes pathway coordination by product binding to avoid the release of cytotoxic and mutagenic intermediates. Similar excision chemistry may apply to other biological reaction pathways requiring the coordination of complex multistep chemical transformations. PMID- 10805772 TI - DNA polymerase active site is highly mutable: evolutionary consequences. AB - DNA polymerases contain active sites that are structurally superimposable and highly conserved in sequence. To assess the significance of this preservation and to determine the mutational burden that active sites can tolerate, we randomly mutated a stretch of 13 amino acids within the polymerase catalytic site (motif A) of Thermus aquaticus DNA polymerase I. After selection, by using genetic complementation, we obtained a library of approximately 8, 000 active mutant DNA polymerases, of which 350 were sequenced and analyzed. This is the largest collection of physiologically active polymerase mutants. We find that all residues of motif A, except one (Asp-610), are mutable while preserving wild-type activity. A wide variety of amino acid substitutions were obtained at sites that are evolutionarily maintained, and conservative substitutions predominate at regions that stabilize tertiary structures. Several mutants exhibit unique properties, including DNA polymerase activity higher than the wild-type enzyme or the ability to incorporate ribonucleotide analogs. Bacteria dependent on these mutated polymerases for survival are fit to replicate repetitively. The high mutability of the polymerase active site in vivo and the ability to evolve altered enzymes may be required for survival in environments that demand increased mutagenesis. The inherent substitutability of the polymerase active site must be addressed relative to the constancy of nucleotide sequence found in nature. PMID- 10805773 TI - Glutathione synthesis is essential for mouse development but not for cell growth in culture. AB - Glutathione (GSH) is a major source of reducing equivalents in mammalian cells. To examine the role of GSH synthesis in development and cell growth, we generated mice deficient in GSH by a targeted disruption of the heavy subunit of gamma glutamylcysteine synthetase (gammaGCS-HS(tm1)), an essential enzyme in GSH synthesis. Embryos homozygous for gammaGCS-HS(tm1) fail to gastrulate, do not form mesoderm, develop distal apoptosis, and die before day 8.5. Lethality results from apoptotic cell death rather than reduced cell proliferation. We also isolated cell lines from homozygous mutant blastocysts in medium containing GSH. These cells also grow indefinitely in GSH-free medium supplemented with N acetylcysteine and have undetectable levels of GSH; further, they show no changes in mitochondrial morphology as judged by electron microscopy. These data demonstrate that GSH is required for mammalian development but dispensable in cell culture and that the functions of GSH, not GSH itself, are essential for cell growth. PMID- 10805774 TI - (4-aminomethyl)phenylguanidine derivatives as nonpeptidic highly selective inhibitors of human urokinase. AB - Increased expression of the serine protease urokinase-type plasminogen activator (uPA) in tumor tissues is highly correlated with tumor cell migration, invasion, proliferation, progression, and metastasis. Thus inhibition of uPA activity represents a promising target for antimetastatic therapy. So far, only the x-ray crystal structure of uPA inactivated by H-Glu-Gly-Arg-chloromethylketone has been reported, thus limited data are available for a rational structure-based design of uPA inhibitors. Taking into account the trypsin-like arginine specificity of uPA, (4-aminomethyl)phenylguanidine was selected as a potential P1 residue and iterative derivatization of its amino group with various hydrophobic residues, and structure-activity relationship-based optimization of the spacer in terms of hydrogen bond acceptor/donor properties led to N-(1-adamantyl)-N'-(4 guanidinobenzyl)urea as a highly selective nonpeptidic uPA inhibitor. The x-ray crystal structure of the uPA B-chain complexed with this inhibitor revealed a surprising binding mode consisting of the expected insertion of the phenylguanidine moiety into the S1 pocket, but with the adamantyl residue protruding toward the hydrophobic S1' enzyme subsite, thus exposing the ureido group to hydrogen-bonding interactions. Although in this enzyme-bound state the inhibitor is crossing the active site, interactions with the catalytic residues Ser-195 and His-57 are not observed, but their side chains are spatially displaced for steric reasons. Compared with other trypsin-like serine proteases, the S2 and S3/S4 pockets of uPA are reduced in size because of the 99-insertion loop. Therefore, the peculiar binding mode of the new type of uPA inhibitors offers the possibility of exploiting optimized interactions at the S1'/S2' subsites to further enhance selectivity and potency. Because crystals of the uPA/benzamidine complex allow inhibitor exchange by soaking procedures, the structure-based design of new generations of uPA inhibitors can rely on the assistance of x-ray analysis. PMID- 10805775 TI - Asparagine-proline sequence within membrane-spanning segment of SREBP triggers intramembrane cleavage by site-2 protease. AB - The NH(2)-terminal domains of membrane-bound sterol regulatory element-binding proteins (SREBPs) are released into the cytosol by regulated intramembrane proteolysis, after which they enter the nucleus to activate genes encoding lipid biosynthetic enzymes. Intramembrane proteolysis is catalyzed by Site-2 protease (S2P), a hydrophobic zinc metalloprotease that cleaves SREBPs at a membrane embedded leucine-cysteine bond. In the current study, we use domain-swapping methods to localize the residues within the SREBP-2 membrane-spanning segment that are required for cleavage by S2P. The studies reveal a requirement for an asparagine-proline sequence in the middle third of the transmembrane segment. We propose a model in which the asparagine-proline sequence serves as an NH(2) terminal cap for a portion of the transmembrane alpha-helix of SREBP, allowing the remainder of the alpha-helix to unwind partially to expose the peptide bond for cleavage by S2P. PMID- 10805776 TI - Assembly of tau protein into Alzheimer paired helical filaments depends on a local sequence motif ((306)VQIVYK(311)) forming beta structure. AB - We have searched for a minimal interaction motif in tau protein that supports the aggregation into Alzheimer-like paired helical filaments. Digestion of the repeat domain with different proteases yields a GluC-induced fragment comprising 43 residues (termed PHF43), which represents the third repeat of tau plus some flanking residues. This fragment self assembles readily into thin filaments without a paired helical appearance, but these filaments are highly competent to nucleate bona fide PHFs from full-length tau. Probing the interactions of PHF43 with overlapping peptides derived from the full tau sequence yields a minimal hexapeptide interaction motif of (306)VQIVYK(311) at the beginning of the third internal repeat. This motif coincides with the highest predicted beta-structure potential in tau. CD and Fourier transform infrared spectroscopy shows that PHF43 acquires pronounced beta structure in conditions of self assembly. Point mutations in the hexapeptide region by proline-scanning mutagenesis prevent the aggregation. The data indicate that PHF assembly is initiated by a short fragment containing the minimal interaction motif forming a local beta structure embedded in a largely random-coil protein. PMID- 10805777 TI - Direct nitric oxide signal transduction via nitrosylation of iron-sulfur centers in the SoxR transcription activator. AB - Nitric oxide (NO) has diverse roles in intercellular communication and (at higher levels) in immune-mediated cell killing. NO reacts with many cellular targets, with cell-killing effects correlated to inactivation of key enzymes through nitrosylation of their iron-sulfur centers. SoxR protein, a redox-sensitive transcription activator dependent on the oxidation state of its binuclear iron sulfur ([2Fe-2S]) centers, is also activated in Escherichia coli on exposure to macrophage-generated NO. We show here that SoxR activation by NO occurs through direct modification of the [2Fe-2S] centers to form protein-bound dinitrosyl-iron dithiol adducts, which we have observed both in intact bacterial cells and in purified SoxR after NO treatment. Functional activation through nitrosylation of iron-sulfur centers contrasts with the inactivation typically caused by this modification. Purified, nitrosylated SoxR has transcriptional activity similar to that of oxidized SoxR and is relatively stable. In contrast, nitrosylated SoxR is short-lived in intact cells, indicative of mechanisms that actively dispose of nitrosylated iron-sulfur centers. PMID- 10805778 TI - Noninvasive measurement of gene expression in skeletal muscle. AB - We have developed a noninvasive detection method for expression of viral-mediated gene transfer. A recombinant adenovirus was constructed by using the gene for arginine kinase (AK), which is the invertebrate correlate to the vertebrate ATP buffering enzyme, creatine kinase. Gene expression was noninvasively monitored using (31)P-magnetic resonance spectroscopy ((31)P-MRS). The product of the AK enzyme, phosphoarginine (PArg), served as an MRS-visible reporter of AK expression. The recombinant adenovirus coding for arginine kinase (rAdCMVAK) was injected into the right hindlimbs of neonatal mice. Two weeks after injection of rAdCMVAK, a unique (31)P-MRS resonance was observed. It was observable in all rAdCMVAK injected hindlimbs and was not present in the contralateral control or the vehicle injected limb. PArg and phosphocreatine (PCr) concentrations were calculated to be 11.6 +/- 0.90 and 13.6 +/- 1.1 mM respectively in rAdCMVAK injected limbs. AK activity was demonstrated in vivo by monitoring the decreases in PArg and ATP resonances during prolonged ischemia. After 1 h of ischemia intracellular pH was 6.73 +/- 0.06, PCr/ATP was decreased by 77 +/- 8%, whereas PArg/ATP was decreased by 50 +/- 15% of basal levels. PArg and PCr returned to basal levels within 5 min of the restoration of blood flow. AK activity persisted for at least 8 mo after injection, indicating that adenoviral-mediated gene transfer can produce stable expression for long periods of time. Therefore, the cDNA encoding AK provides a useful reporter gene that allows noninvasive and repeated monitoring of gene expression after viral mediated gene transfer to muscle. PMID- 10805779 TI - Detection and selective dissociation of intact ribosomes in a mass spectrometer. AB - Intact Escherichia coli ribosomes have been projected into the gas phase of a mass spectrometer by means of nanoflow electrospray techniques. Species with mass/charge ratios in excess of 20,000 were detected at the level of individual ions by using time-of-flight analysis. Once in the gas phase the stability of intact ribosomes was investigated and found to increase as a result of cross linking ribosomal proteins to the rRNA. By lowering the Mg(2+) concentration in solutions containing ribosomes the particles were found to dissociate into 30S and 50S subunits. The resolution of the charge states in the spectrum of the 30S subunit enabled its mass to be determined as 852,187 +/- 3,918 Da, a value within 0.6% of that calculated from the individual proteins and the 16S RNA. Further dissociation into smaller macromolecular complexes and then individual proteins could be induced by subjecting the particles to increasingly energetic gas phase collisions. The ease with which proteins dissociated from the intact species was found to be related to their known interactions in the ribosome particle. The results show that emerging mass spectrometric techniques can be used to characterize a fully functional biological assembly as well as its isolated components. PMID- 10805780 TI - A green fluorescent protein-reporter mammalian two-hybrid system with extrachromosomal maintenance of a prey expression plasmid: application to interaction screening. AB - An improved mammalian two-hybrid system designed for interaction trap screening is described in this paper. CV-1/EBNA-1 monkey kidney epithelial cells expressing Epstein-Barr virus nuclear antigen 1 (EBNA-1) were stably transfected with a reporter plasmid for GAL4-dependent expression of the green fluorescent protein (GFP). A resulting clone, GB133, expressed GFP strongly when transfected transiently with transcriptional activators fused to GAL4 DNA-binding domain with minimal background GFP expression. GB133 cells maintained plasmids containing the OriP Epstein-Barr virus replication origin that directs replication of plasmids in mammalian cells in the presence of the EBNA-1 protein. GB133 cells transfected stably with a model bait expressed GFP when further transfected transiently with an expression plasmid for a known positive prey. When the bait-expressing GB133 cells were transfected transiently with an OriP-containing expression plasmid for the positive prey together with excess amounts of empty vector, cells that received the positive prey were readily identified by green fluorescence in cell culture and eventually formed green fluorescent microcolonies, because the prey plasmid was maintained by the EBNA-1/Ori-P system. The green fluorescent microcolonies were harvested directly from the culture dishes under a fluorescence microscope, and total DNA was then prepared. Prey-encoding cDNA was recovered by PCR using primers annealing to the vector sequences flanking the insert-cloning site. This system should be useful in mammalian cells for efficient screening of cDNA libraries by two-hybrid interaction. PMID- 10805781 TI - Role of Rac in controlling the actin cytoskeleton and chemotaxis in motile cells. AB - We have used the chemotactic ability of Dictyostelium cells to examine the roles of Rho family members, known regulators of the assembly of F-actin, in cell movement. Wild-type cells polarize with a leading edge enriched in F-actin toward a chemoattractant. Overexpression of constitutively active Dictyostelium Rac1B(61L) or disruption of DdRacGAP1, which encodes a Dictyostelium Rac1 GAP, induces membrane ruffles enriched with actin filaments around the perimeter of the cell and increased levels of F-actin in resting cells. Whereas wild-type cells move linearly toward the cAMP source, Rac1B(61L) and Ddracgap1 null cells make many wrong turns and chemotaxis is inefficient, which presumably results from the unregulated activation of F-actin assembly and pseudopod extension. Cells expressing dominant-negative DdRac1B(17N) do not have a well-defined F actin-rich leading edge and do not protrude pseudopodia, resulting in very poor cell motility. From these studies and assays examining chemoattractant-mediated F actin assembly, we suggest DdRac1 regulates the basal levels of F-actin assembly, its dynamic reorganization in response to chemoattractants, and cellular polarity during chemotaxis. PMID- 10805782 TI - NMR and mutagenesis evidence for an I domain allosteric site that regulates lymphocyte function-associated antigen 1 ligand binding. AB - The leukocyte integrin, lymphocyte function-associated antigen 1 (LFA-1) (CD11a/CD18), mediates cell adhesion and signaling in inflammatory and immune responses. To support these functions, LFA-1 must convert from a resting to an activated state that avidly binds its ligands such as intercellular adhesion molecule 1 (ICAM-1). Biochemical and x-ray studies of the Mac-1 (CD11b/CD18) I domain suggest that integrin activation could involve a conformational change of the C-terminal alpha-helix. We report the use of NMR spectroscopy to identify CD11a I domain residues whose resonances are affected by binding to ICAM-1. We observed two distinct sites in the CD11a I domain that were affected. As expected from previous mutagenesis studies, a cluster of residues localized around the metal ion-dependent adhesion site (MIDAS) was severely perturbed on ICAM-1 binding. A second cluster of residues distal to the MIDAS that included the C terminal alpha-helix of the CD11a I domain was also affected. Substitution of residues in the core of this second I domain site resulted in constitutively active LFA-1 binding to ICAM-1. Binding data indicates that none of the 20 substitution mutants we tested at this second site form an essential ICAM-1 binding interface. We also demonstrate that residues in the I domain linker sequences can regulate LFA-1 binding. These results indicate that LFA-1 binding to ICAM-1 is regulated by an I domain allosteric site (IDAS) and that this site is structurally linked to the MIDAS. PMID- 10805783 TI - Non-CpG methylation is prevalent in embryonic stem cells and may be mediated by DNA methyltransferase 3a. AB - Current evidence indicates that methylation of cytosine in mammalian DNA is restricted to both strands of the symmetrical sequence CpG, although there have been sporadic reports that sequences other than CpG may also be methylated. We have used a dual-labeling nearest neighbor technique and bisulphite genomic sequencing methods to investigate the nearest neighbors of 5-methylcytosine residues in mammalian DNA. We find that embryonic stem cells, but not somatic tissues, have significant cytosine-5 methylation at CpA and, to a lesser extent, at CpT. As the expression of the de novo methyltransferase Dnmt3a correlates well with the presence of non-CpG methylation, we asked whether Dnmt3a might be responsible for this modification. Analysis of genomic methylation in transgenic Drosophila expressing Dnmt3a reveals that Dnmt3a is predominantly a CpG methylase but also is able to induce methylation at CpA and at CpT. PMID- 10805784 TI - MEK kinase 1 is critically required for c-Jun N-terminal kinase activation by proinflammatory stimuli and growth factor-induced cell migration. AB - Exposure of eukaryotic cells to extracellular stimuli results in activation of mitogen-activated protein kinase (MAPK) cascades composed of MAPKs, MAPK kinases (MAP2Ks), and MAPK kinase kinases (MAP3Ks). Mammals possess a large number of MAP3Ks, many of which can activate the c-Jun N-terminal kinase (JNK) MAPK cascade when overexpressed, but whose biological function is poorly understood. We examined the function of the MAP3K MEK kinase 1 (MEKK1) in proinflammatory signaling. Using MEKK1-deficient embryonic stem cells prepared by gene targeting, we find that, in addition to its function in JNK activation by growth factors, MEKK1 is required for JNK activation by diverse proinflammatory stimuli, including tumor necrosis factor alpha, IL-1, double-stranded RNA, and lipopolysaccharide. MEKK1 is also essential for induction of embryonic stem cell migration by serum factors, but is not required for activation of other MAPKs or the IkappaB kinase signaling cascade. PMID- 10805785 TI - Sid4p is required to localize components of the septation initiation pathway to the spindle pole body in fission yeast. AB - A mutation in the Schizosaccharomyces pombe sid4(+) (septation initiation defective) gene was isolated in a screen for mutants defective in cytokinesis. We have cloned sid4(+) and have found that sid4(+) encodes a previously unknown 76.4 kDa protein that localizes to the spindle pole body (SPB) throughout the cell cycle. Sid4p is required for SPB localization of key regulators of septation initiation, including the GTPase Spg1p, the protein kinase Cdc7p, and the GTPase activating protein Byr4p. An N-terminally truncated Sid4p mutant does not localize to SPBs and when overproduced acts as a dominant-negative mutant by titrating endogenous Sid4p and Spg1p from the SPB. Conversely, the Sid4p N terminal 153 amino acids are sufficient for SPB localization. Biochemical studies demonstrate that Sid4p interacts with itself, and yeast two-hybrid analysis shows that its self-interaction domain lies within the C-terminal half of the protein. Our data indicate that Sid4p SPB localization is a prerequisite for the execution of the Spg1p signaling cascade. PMID- 10805786 TI - Tissue factor- and factor X-dependent activation of protease-activated receptor 2 by factor VIIa. AB - Protease-activated receptor 2 (PAR2) is expressed by vascular endothelial cells and other cells in which its function and physiological activator(s) are unknown. Unlike PAR1, PAR3, and PAR4, PAR2 is not activatable by thrombin. Coagulation factors VIIa (FVIIa) and Xa (FXa) are proteases that act upstream of thrombin in the coagulation cascade and require cofactors to interact with their substrates. These proteases elicit cellular responses, but their receptor(s) have not been identified. We asked whether FVIIa and FXa might activate PARs if presented by their cofactors. Co-expression of tissue factor (TF), the cellular cofactor for FVIIa, together with PAR1, PAR2, PAR3, or PAR4 conferred TF-dependent FVIIa activation of PAR2 and, to lesser degree, PAR1. Responses to FXa were also observed but were independent of exogenous cofactor. The TF/FVIIa complex converts the inactive zymogen Factor X (FX) to FXa. Strikingly, when FX was present, low picomolar concentrations of FVIIa caused robust signaling in cells expressing TF and PAR2. Responses in keratinocytes and cytokine-treated endothelial cells suggested that PAR2 may be activated directly by TF/FVIIa and indirectly by TF/FVIIa-generated FXa at naturally occurring expression levels of TF and PAR2. These results suggest that PAR2, although not activatable by thrombin, may nonetheless function as a sensor for coagulation proteases and contribute to endothelial activation in the setting of injury and inflammation. More generally, these findings highlight the potential importance of cofactors in regulating PAR function and specificity. PMID- 10805787 TI - Distinct roles of the NH2- and COOH-terminal domains of the protein inhibitor of activated signal transducer and activator of transcription (STAT) 1 (PIAS1) in cytokine-induced PIAS1-Stat1 interaction. AB - STATs are activated by tyrosine phosphorylation on cytokine stimulation. A tyrosine-phosphorylated STAT forms a functional dimer through reciprocal Src homology 2 domain (SH2)-phosphotyrosyl peptide interactions. IFN treatment induces the association of PIAS1 and Stat1, which results in the inhibition of Stat1-mediated gene activation. The molecular basis of the cytokine-dependent PIAS1-Stat1 interaction has not been understood. We report here that a region near the COOH terminus of PIAS1 (amino acids 392-541) directly interacts with the NH(2)-terminal domain of Stat1 (amino acids 1-191). A mutant PIAS1 lacking the Stat1-interacting domain failed to inhibit Stat1-mediated gene activation. By using a modified yeast two-hybrid assay, we demonstrated that PIAS1 specifically interacts with the Stat1 dimer, but not tyrosine-phosphorylated or unphosphorylated Stat1 monomer. In addition, whereas the NH(2)-terminal region of PIAS1 does not interact with Stat1, it serves as a modulatory domain by preventing the interaction of the COOH-terminal domain of PIAS1 with the Stat1 monomer. Thus, the cytokine-induced PIAS1-Stat1 interaction is mediated through the specific recognition of the dimeric form of Stat1 by PIAS1. PMID- 10805788 TI - Somatic polyploidization and cellular proliferation drive body size evolution in nematodes. AB - Most of the hypodermis of a rhabditid nematode such as Caenorhabditis elegans is a single syncytium. The size of this syncytium (as measured by body size) has evolved repeatedly in the rhabditid nematodes. Two cellular mechanisms are important in the evolution of body size: changes in the numbers of cells that fuse with the syncytium, and the extent of its acellular growth. Thus nematodes differ from mammals and other invertebrates in which body size evolution is caused by changes in cell number alone. The evolution of acellular syncytial growth in nematodes is also associated with changes in the ploidy of hypodermal nuclei. These nuclei are polyploid as a consequence of iterative rounds of endoreduplication, and this endocycle has evolved repeatedly. The association between acellular growth and endoreduplication is also seen in C. elegans mutations that interrupt transforming growth factor-beta signaling and that result in dwarfism and deficiencies in hypodermal ploidy. The transforming growth factor-beta pathway is a candidate for being involved in nematode body size evolution. PMID- 10805789 TI - Novel gene conversion between X-Y homologues located in the nonrecombining region of the Y chromosome in Felidae (Mammalia). AB - Genes located on the mammalian Y chromosome outside of the pseudoautosomal region do not recombine with those on the X and are predicted to either undergo selection for male function or gradually degenerate because of an accumulation of deleterious mutations. Here, phylogenetic analyses of X-Y homologues, Zfx and Zfy, among 26 felid species indicate two ancestral episodes of directed genetic exchange (ectopic gene conversion) from X to Y: once during the evolution of pallas cat and once in a common predecessor of ocelot lineage species. Replacement of the more rapidly evolving Y homologue with the evolutionarily constrained X copy may represent a mechanism for adaptive editing of functional genes on the nonrecombining region of the mammalian Y chromosome. PMID- 10805790 TI - Permanence or change? The meaning of genetic variation. AB - Selected aspects of the evolutionary process and more specifically of the genetic variation are considered, with an emphasis in studies performed by my group. One key aspect of evolution seems to be the concomitant occurrence of dichotomic, contradictory (dialect) processes. Genetic variation is structured, and the dynamics of change at one level is not necessarily paralleled by that in another. The pathogenesis-related protein superfamily can be cited as an example in which permanence (the maintenance of certain key genetic features) coexists with change (modifications that led to different functions in different classes of organisms). Relationships between structure and function are exemplified by studies with hemoglobin Porto Alegre. The genetic structure of tribal populations may differ in important aspects from that of industrialized societies. Evolutionary histories also may differ when considered through the investigation of patrilineal or matrilineal lineages. Global evaluations taking into consideration all of these aspects are needed if we really want to understand the meaning of genetic variation. PMID- 10805791 TI - Rapid evolution in plant chitinases: molecular targets of selection in plant pathogen coevolution. AB - Many pathogen recognition genes, such as plant R-genes, undergo rapid adaptive evolution, providing evidence that these genes play a critical role in plant pathogen coevolution. Surprisingly, whether rapid adaptive evolution also occurs in genes encoding other kinds of plant defense proteins is unknown. Unlike recognition proteins, plant chitinases attack pathogens directly, conferring disease resistance by degrading chitin, a component of fungal cell walls. Here, we show that nonsynonymous substitution rates in plant class I chitinase often exceed synonymous rates in the plant genus Arabis (Cruciferae) and in other dicots, indicating a succession of adaptively driven amino acid replacements. We identify individual residues that are likely subject to positive selection by using codon substitution models and determine the location of these residues on the three-dimensional structure of class I chitinase. In contrast to primate lysozymes and plant class III chitinases, structural and functional relatives of class I chitinase, the adaptive replacements of class I chitinase occur disproportionately in the active site cleft. This highly unusual pattern of replacements suggests that fungi directly defend against chitinolytic activity through enzymatic inhibition or other forms of chemical resistance and identifies target residues for manipulating chitinolytic activity. These data also provide empirical evidence that plant defense proteins not involved in pathogen recognition also evolve in a manner consistent with rapid coevolutionary interactions. PMID- 10805792 TI - An ancestral MADS-box gene duplication occurred before the divergence of plants and animals. AB - Changes in genes encoding transcriptional regulators can alter development and are important components of the molecular mechanisms of morphological evolution. MADS-box genes encode transcriptional regulators of diverse and important biological functions. In plants, MADS-box genes regulate flower, fruit, leaf, and root development. Recent sequencing efforts in Arabidopsis have allowed a nearly complete sampling of the MADS-box gene family from a single plant, something that was lacking in previous phylogenetic studies. To test the long-suspected parallel between the evolution of the MADS-box gene family and the evolution of plant form, a polarized gene phylogeny is necessary. Here we suggest that a gene duplication ancestral to the divergence of plants and animals gave rise to two main lineages of MADS-box genes: TypeI and TypeII. We locate the root of the eukaryotic MADS-box gene family between these two lineages. A novel monophyletic group of plant MADS domains (AGL34 like) seems to be more closely related to previously identified animal SRF-like MADS domains to form TypeI lineage. Most other plant sequences form a clear monophyletic group with animal MEF2-like domains to form TypeII lineage. Only plant TypeII members have a K domain that is downstream of the MADS domain in most plant members previously identified. This suggests that the K domain evolved after the duplication that gave rise to the two lineages. Finally, a group of intermediate plant sequences could be the result of recombination events. These analyses may guide the search for MADS-box sequences in basal eukaryotes and the phylogenetic placement of new genes from other plant species. PMID- 10805793 TI - Gene content phylogeny of herpesviruses. AB - Clusters of orthologous groups [COGs; Tatusov, R. L., Koonin, E. V. & Lipman, D. J. (1997) Science 278, 631-637] were identified for a set of 13 completely sequenced herpesviruses. Each COG represented a family of gene products conserved across several herpes genomes. These families were defined without using an arbitrary threshold criterion based on sequence similarity. The COG technique was modified so that variable stringency in COG construction was possible. High stringencies identify a core set of highly conserved genes. Varying COG stringency reveals differences in the degree of conservation between functional classes of genes. The COG data were used to construct whole-genome phylogenetic trees based on gene content. These trees agree well with trees based on other methods and are robust when tested by bootstrap analysis. The COG data also were used to construct a reciprocal tree that clustered genes with similar phylogenetic profiles. This clustering may give clues to genes with related functions or with related histories of acquisition and loss during herpesvirus evolution. PMID- 10805794 TI - Behavioral alterations associated with apoptosis and down-regulation of presenilin 1 in the brains of p53-deficient mice. AB - Presenilin 1 (PS1) expression is repressed by the p53 tumor suppressor. As shown herein, wild-type PS1 is an effective antiapoptotic molecule capable of significantly inhibiting p53-dependent and p53-independent cell death. We analyzed, at the functional and molecular levels, the brains of p53 knockout mice. Surprisingly, we found that lack of p53 expression induces apoptotic brain lesions, accompanied by learning deficiency and behavioral alterations. p53 deficient mice show an unexpected overexpression of p21(waf1) with subsequent down-regulation of PS1 in their brains. This process is progressive and age dependent. These data indicate that the p53 pathway, besides affecting tumor suppression, may play a major role in regulating neurobehavioral function and cell survival in the brain. PMID- 10805795 TI - LexA chimeras reveal the function of Drosophila Fos as a context-dependent transcriptional activator. AB - The transcriptional activation potential of proteins can be assayed in chimeras containing a heterologous DNA-binding domain that mediates their recruitment to reporter genes. This approach has been widely used in yeast and in transient mammalian cell assays. Here, we applied it to assay the transactivation potential of proteins in transgenic Drosophila embryos. We found that a chimera between the DNA-binding bacterial LexA protein and the transactivation domain from yeast GAL4 behaved as a potent synthetic activator in all embryonic tissues. In contrast, a LexA chimera containing Drosophila Fos (Dfos) required an unexpected degree of context to function as a transcriptional activator. We provide evidence to suggest that this context is provided by Djun and Mad (a Drosophila Smad), and that these partner factors need to be activated by signaling from Jun N-terminal kinase and decapentaplegic, respectively. Because Dfos behaves as an autonomous transcriptional activator in more artificial assays systems, our data suggest that context-dependence of transcription factors may be more prevalent than previously thought. PMID- 10805796 TI - Chromosomal breakage-fusion-bridge events cause genetic intratumor heterogeneity. AB - It has long been known that rearrangements of chromosomes through breakage-fusion bridge (BFB) cycles may cause variability of phenotypic and genetic traits within a cell population. Because intercellular heterogeneity is often found in neoplastic tissues, we investigated the occurrence of BFB events in human solid tumors. Evidence of frequent BFB events was found in malignancies that showed unspecific chromosome aberrations, including ring chromosomes, dicentric chromosomes, and telomeric associations, as well as extensive intratumor heterogeneity in the pattern of structural changes but not in tumors with tumor specific aberrations and low variability. Fluorescence in situ hybridization analysis demonstrated that chromosomes participating in anaphase bridge formation were involved in a significantly higher number of structural aberrations than other chromosomes. Tumors with BFB events showed a decreased elimination rate of unstable chromosome aberrations after irradiation compared with normal cells and other tumor cells. This result suggests that a combination of mitotically unstable chromosomes and an elevated tolerance to chromosomal damage leads to constant genomic reorganization in many malignancies, thereby providing a flexible genetic system for clonal evolution and progression. PMID- 10805797 TI - Rescue of dystrophin expression in mdx mouse muscle by RNA/DNA oligonucleotides. AB - Chimeric RNA/DNA oligonucleotides ("chimeraplasts") have been shown to induce single base alterations in genomic DNA both in vitro and in vivo. The mdx mouse strain has a point mutation in the dystrophin gene, the consequence of which is a muscular dystrophy resulting from deficiency of the dystrophin protein in skeletal muscle. To test the feasibility of chimeraplast-mediated gene therapy for muscular dystrophies, we used a chimeraplast (designated "MDX1") designed to correct the point mutation in the dystrophin gene in mdx mice. After direct injection of MDX1 into muscles of mdx mice, immunohistochemical analysis revealed dystrophin-positive fibers clustered around the injection site. Two weeks after single injections into tibialis anterior muscles, the maximum number of dystrophin-positive fibers (approximately 30) in any muscle represented 1-2% of the total number of fibers in that muscle. Ten weeks after single injections, the range of the number of dystrophin-positive fibers was similar to that seen after 2 wk, suggesting that the expression was stable, as would be predicted for a gene conversion event. Staining with exon-specific antibodies showed that none of these were "revertant fibers." Furthermore, dystrophin from MDX1-injected muscles was full length by immunoblot analysis. No dystrophin was detectable by immunohistochemical or immunoblot analysis after control chimeraplast injections. Finally, reverse transcription-PCR analysis demonstrated the presence of transcripts with the wild-type dystrophin sequence only in mdx muscles injected with MDX1 chimeraplasts. These results provide the foundation for further studies of chimeraplast-mediated gene therapy as a therapeutic approach to muscular dystrophies and other genetic disorders of muscle. PMID- 10805798 TI - Long-term kinetics of T cell production in HIV-infected subjects treated with highly active antiretroviral therapy. AB - The long-term kinetics of T cell production following highly active antiretroviral therapy (HAART) were investigated in blood and lymph node in a group of HIV-infected subjects at early stage of established infection and prospectively studied for 72 wk. Before HAART, CD4 and CD8 T cell turnover was increased. However, the total number of proliferating CD4(+) T lymphocytes, i.e., CD4(+)Ki67(+) T lymphocytes, was not significantly different in HIV-infected (n = 73) and HIV-negative (n = 15) subjects, whereas proliferating CD8(+)Ki67(+) T lymphocytes were significantly higher in HIV-infected subjects. After HAART, the total body number of proliferating CD4(+)Ki67(+) T lymphocytes increased over time and was associated with an increase of both naive and memory CD4(+) T cells. The maximal increase (2-fold) was observed at week 36, whereas at week 72 the number of proliferating CD4(+) T cells dropped to baseline levels, i.e., before HAART. The kinetics of the fraction of proliferating CD4 and CD8 T cells were significantly correlated with the changes in the total body number of these T cell subsets. These results demonstrate a direct relationship between ex vivo measures of T cell production and quantitative changes in total body T lymphocyte populations. This study provides advances in the delineation of the kinetics of T cell production in HIV infection in the presence and/or in the absence of HAART. PMID- 10805799 TI - Crystal structure of a Staphylococcus aureus protein A domain complexed with the Fab fragment of a human IgM antibody: structural basis for recognition of B-cell receptors and superantigen activity. AB - Staphylococcus aureus produces a virulence factor, protein A (SpA), that contains five homologous Ig-binding domains. The interactions of SpA with the Fab region of membrane-anchored Igs can stimulate a large fraction of B cells, contributing to lymphocyte clonal selection. To understand the molecular basis for this activity, we have solved the crystal structure of the complex between domain D of SpA and the Fab fragment of a human IgM antibody to 2.7-A resolution. In the complex, helices II and III of domain D interact with the variable region of the Fab heavy chain (V(H)) through framework residues, without the involvement of the hypervariable regions implicated in antigen recognition. The contact residues are highly conserved in human V(H)3 antibodies but not in other families. The contact residues from domain D also are conserved among all SpA Ig-binding domains, suggesting that each could bind in a similar manner. Features of this interaction parallel those reported for staphylococcal enterotoxins that are superantigens for many T cells. The structural homology between Ig V(H) regions and the T-cell receptor V(beta) regions facilitates their comparison, and both types of interactions involve lymphocyte receptor surface remote from the antigen binding site. However, T-cell superantigens reportedly interact through hydrogen bonds with T-cell receptor V(beta) backbone atoms in a primary sequence-independent manner, whereas SpA relies on a sequence-restricted conformational binding with residue side chains, suggesting that this common bacterial pathogen has adopted distinct molecular recognition strategies for affecting large sets of B and T lymphocytes. PMID- 10805800 TI - D-beta-hydroxybutyrate protects neurons in models of Alzheimer's and Parkinson's disease. AB - The heroin analogue 1-methyl-4-phenylpyridinium, MPP(+), both in vitro and in vivo, produces death of dopaminergic substantia nigral cells by inhibiting the mitochondrial NADH dehydrogenase multienzyme complex, producing a syndrome indistinguishable from Parkinson's disease. Similarly, a fragment of amyloid protein, Abeta(1-42), is lethal to hippocampal cells, producing recent memory deficits characteristic of Alzheimer's disease. Here we show that addition of 4 mM d-beta-hydroxybutyrate protected cultured mesencephalic neurons from MPP(+) toxicity and hippocampal neurons from Abeta(1-42) toxicity. Our previous work in heart showed that ketone bodies, normal metabolites, can correct defects in mitochondrial energy generation. The ability of ketone bodies to protect neurons in culture suggests that defects in mitochondrial energy generation contribute to the pathophysiology of both brain diseases. These findings further suggest that ketone bodies may play a therapeutic role in these most common forms of human neurodegeneration. PMID- 10805801 TI - Molecular basis for a link between complement and the vascular complications of diabetes. AB - Activated terminal complement proteins C5b to C9 form the membrane attack complex (MAC) pore. Insertion of the MAC into endothelial cell membranes causes the release of growth factors that stimulate tissue growth and proliferation. The complement regulatory membrane protein CD59 restricts MAC formation. Because increased cell proliferation characterizes the major chronic vascular complications of human diabetes and because increased glucose levels in diabetes cause protein glycation and impairment of protein function, we investigated whether glycation could inhibit CD59. Glycation-inactivation of CD59 would cause increased MAC deposition and MAC-stimulated cell proliferation. Here, we report that (i) human CD59 is glycated in vivo, (ii) glycated human CD59 loses its MAC inhibitory function, and (iii) inactivation of CD59 increases MAC-induced growth factor release from endothelial cells. We demonstrate by site-directed mutagenesis that residues K41 and H44 form a preferential glycation motif in human CD59. The presence of this glycation motif in human CD59, but not in CD59 of other species, may help explain the distinct propensity of humans to develop vascular proliferative complications of diabetes. PMID- 10805802 TI - AIS is an oncogene amplified in squamous cell carcinoma. AB - We and others recently isolated a human p53 homologue (p40/p51/p63/p73L) and localized the gene to the distal long arm of chromosome 3. Here we sought to examine the role of p40/p73L, two variants lacking the N-terminal transactivation domain, in cancer. Fluorescent in situ hybridization (FISH) analysis revealed frequent amplification of this gene locus in primary squamous cell carcinoma of the lung and head and neck cancer cell lines. (We named this locus AIS for amplified in squamous cell carcinoma.) Furthermore, amplification of the AIS locus was accompanied by RNA and protein overexpression of a variant p68(AIS) lacking the terminal transactivation domain. Protein overexpression in primary lung tumors was limited to squamous cell carcinoma and tumors known to harbor a high frequency of p53 mutations. Overexpression of p40(AIS) in Rat 1a cells led to an increase in soft agar growth and tumor size in mice. Our results support the idea that AIS plays an oncogenic role in human cancer. PMID- 10805803 TI - Ultrasensitive detection of pathological prion protein aggregates by dual-color scanning for intensely fluorescent targets. AB - A definite diagnosis of prion diseases such as Creutzfeldt-Jakob disease (CJD) relies on the detection of pathological prion protein (PrP(Sc)). However, no test for PrP(Sc) in cerebrospinal fluid (CSF) has been available thus far. Based on a setup for confocal dual-color fluorescence correlation spectroscopy, a technique suitable for single molecule detection, we developed a highly sensitive detection method for PrP(Sc). Pathological prion protein aggregates were labeled by specific antibody probes tagged with fluorescent dyes, resulting in intensely fluorescent targets, which were measured by dual-color fluorescence intensity distribution analysis in a confocal scanning setup. In a diagnostic model system, PrP(Sc) aggregates were detected down to a concentration of 2 pM PrP(Sc), corresponding to an aggregate concentration of approximately 2 fM, which was more than one order of magnitude more sensitive than Western blot analysis. A PrP(Sc) specific signal could also be detected in a number of CSF samples from patients with CJD but not in control samples, providing the basis for a rapid and specific test for CJD and other prion diseases. Furthermore, this method could be adapted to the sensitive detection of other disease-associated amyloid aggregates such as in Alzheimer's disease. PMID- 10805804 TI - Estrogen receptor specificity in the regulation of skeletal growth and maturation in male mice. AB - Androgens may regulate the male skeleton directly through a stimulation of androgen receptors or indirectly through aromatization of androgens into estrogen and, thereafter, through stimulation of estrogen receptors (ERs). The relative importance of ER subtypes in the regulation of the male skeleton was studied in ERalpha-knockout (ERKO), ERbeta-knockout (BERKO), and double ERalpha/beta knockout (DERKO) mice. ERKO and DERKO, but not BERKO, demonstrated decreased longitudinal as well as radial skeletal growth associated with decreased serum levels of insulin-like growth factor I. Therefore, ERalpha, but not ERbeta, mediates important effects of estrogen in the skeleton of male mice during growth and maturation. PMID- 10805805 TI - c-Abl is required for development and optimal cell proliferation in the context of p53 deficiency. AB - The c-Abl tyrosine kinase and the p53 tumor suppressor protein interact functionally and biochemically in cellular genotoxic stress response pathways and are implicated as downstream mediators of ATM (ataxia-telangiectasia mutated). This fact led us to study genetic interactions in vivo between c-Abl and p53 by examining the phenotype of mice and cells deficient in both proteins. c-Abl-null mice show high neonatal mortality and decreased B lymphocytes, whereas p53-null mice are prone to tumor development. Surprisingly, mice doubly deficient in both c-Abl and p53 are not viable, suggesting that c-Abl and p53 together contribute to an essential function required for normal development. Fibroblasts lacking both c-Abl and p53 were similar to fibroblasts deficient in p53 alone, showing loss of the G(1)/S cell-cycle checkpoint and similar clonogenic survival after ionizing radiation. Fibroblasts deficient in both c-Abl and p53 show reduced growth in culture, as manifested by reduction in the rate of proliferation, saturation density, and colony formation, compared with fibroblasts lacking p53 alone. This defect could be restored by reconstitution of c-Abl expression. Taken together, these results indicate that the ATM phenotype cannot be explained solely by loss of c-Abl and p53 and that c-Abl contributes to enhanced proliferation of p53-deficient cells. Inhibition of c-Abl function may be a therapeutic strategy to target p53-deficient cells selectively. PMID- 10805806 TI - Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. AB - Many gram-positive bacteria covalently tether their surface adhesins to the cell wall peptidoglycan. We find that surface proteins of Staphylococcus aureus are linked to the cell wall by sortase, an enzyme that cleaves polypeptides at a conserved LPXTG motif. S. aureus mutants lacking sortase fail to process and display surface proteins and are defective in the establishment of infections. Thus, the cell wall envelope of gram-positive bacteria represents a surface organelle responsible for interactions with the host environment during the pathogenesis of bacterial infections. PMID- 10805807 TI - Engineering the largest RNA virus genome as an infectious bacterial artificial chromosome. AB - The construction of cDNA clones encoding large-size RNA molecules of biological interest, like coronavirus genomes, which are among the largest mature RNA molecules known to biology, has been hampered by the instability of those cDNAs in bacteria. Herein, we show that the application of two strategies, cloning of the cDNAs into a bacterial artificial chromosome and nuclear expression of RNAs that are typically produced within the cytoplasm, is useful for the engineering of large RNA molecules. A cDNA encoding an infectious coronavirus RNA genome has been cloned as a bacterial artificial chromosome. The rescued coronavirus conserved all of the genetic markers introduced throughout the sequence and showed a standard mRNA pattern and the antigenic characteristics expected for the synthetic virus. The cDNA was transcribed within the nucleus, and the RNA translocated to the cytoplasm. Interestingly, the recovered virus had essentially the same sequence as the original one, and no splicing was observed. The cDNA was derived from an attenuated isolate that replicates exclusively in the respiratory tract of swine. During the engineering of the infectious cDNA, the spike gene of the virus was replaced by the spike gene of an enteric isolate. The synthetic virus replicated abundantly in the enteric tract and was fully virulent, demonstrating that the tropism and virulence of the recovered coronavirus can be modified. This demonstration opens up the possibility of employing this infectious cDNA as a vector for vaccine development in human, porcine, canine, and feline species susceptible to group 1 coronaviruses. PMID- 10805808 TI - The Escherichia coli MG1655 in silico metabolic genotype: its definition, characteristics, and capabilities. AB - The Escherichia coli MG1655 genome has been completely sequenced. The annotated sequence, biochemical information, and other information were used to reconstruct the E. coli metabolic map. The stoichiometric coefficients for each metabolic enzyme in the E. coli metabolic map were assembled to construct a genome-specific stoichiometric matrix. The E. coli stoichiometric matrix was used to define the system's characteristics and the capabilities of E. coli metabolism. The effects of gene deletions in the central metabolic pathways on the ability of the in silico metabolic network to support growth were assessed, and the in silico predictions were compared with experimental observations. It was shown that based on stoichiometric and capacity constraints the in silico analysis was able to qualitatively predict the growth potential of mutant strains in 86% of the cases examined. Herein, it is demonstrated that the synthesis of in silico metabolic genotypes based on genomic, biochemical, and strain-specific information is possible, and that systems analysis methods are available to analyze and interpret the metabolic phenotype. PMID- 10805809 TI - Production of infectious bovine papillomavirus from cloned viral DNA by using an organotypic raft/xenograft technique. AB - Bovine papillomavirus type 1 (BPV-1) induces fibropapillomas in its natural host and can transform fibroblasts in culture. The viral genome is maintained as an episome within fibroblasts, which has allowed extensive genetic analyses of the viral functions required for DNA replication, gene expression, and transformation. Much less is known about BPV-1 gene expression and replication in bovine epithelial cells because the study of the complete viral life cycle requires an experimental system capable of generating a fully differentiated stratified bovine epithelium. Using a combination of organotypic raft cultures and xenografts on nude mice, we have developed a system in which BPV-1 can replicate and produce infectious viral particles. Organotypic cultures were established with bovine keratinocytes plated on a collagen raft containing BPV-1 transformed fibroblasts. These keratinocytes were infected with virus particles isolated from a bovine wart or were transfected with cloned BPV-1 DNA. Several days after the rafts were lifted to the air interface, they were grafted on nude mice. After 6-8 weeks, large xenografts were produced that exhibited a hyperplastic and hyperkeratotic epithelium overlying a large dermal fibroma. These lesions were strikingly similar to a fibropapilloma caused by BPV-1 in the natural host. Amplified viral DNA and capsid antigens were detected in the suprabasal cells of the epithelium. Moreover, infectious virus particles could be isolated from these lesions and quantitated by a focus formation assay on mouse cells in culture. Interestingly, analysis of grafts produced with infected and uninfected fibroblasts indicated that the fibroma component was not required for productive infection or morphological changes characteristic of papillomavirus infected epithelium. This system will be a powerful tool for the genetic analysis of the roles of the viral gene products in the complete viral life cycle. PMID- 10805810 TI - Neonatal lead exposure impairs development of rodent barrel field cortex. AB - Childhood exposure to low-level lead can permanently reduce intelligence, but the neurobiologic mechanism for this effect is unknown. We examined the impact of lead exposure on the development of cortical columns, using the rodent barrel field as a model. In all areas of mammalian neocortex, cortical columns constitute a fundamental structural unit subserving information processing. Barrel field cortex contains columnar processing units with distinct clusters of layer IV neurons that receive sensory input from individual whiskers. In this study, rat pups were exposed to 0, 0.2, 1, 1.5, or 2 g/liter lead acetate in their dam's drinking water from birth through postnatal day 10. This treatment, which coincides with the development of segregated columns in the barrel field, produced blood lead concentrations from 1 to 31 microg/dl. On postnatal day 10, the area of the barrel field and of individual barrels was measured. A dose related reduction in barrel field area was observed (Pearson correlation = 0.740; P < 0.001); mean barrel field area in the highest exposure group was decreased 12% versus controls. Individual barrels in the physiologically more active caudoventral group were affected preferentially. Total cortical area measured in the same sections was not altered significantly by lead exposure. These data support the hypothesis that lead exposure may impair the development of columnar processing units in immature neocortex. We demonstrate that low levels of blood lead, in the range seen in many impoverished inner-city children, cause structural alterations in a neocortical somatosensory map. PMID- 10805811 TI - A deletion in a photoreceptor-specific nuclear receptor mRNA causes retinal degeneration in the rd7 mouse. AB - The rd7 mouse, an animal model for hereditary retinal degeneration, has some characteristics similar to human flecked retinal disorders. Here we report the identification of a deletion in a photoreceptor-specific nuclear receptor (mPNR) mRNA that is responsible for hereditary retinal dysplasia and degeneration in the rd7 mouse. mPNR was isolated from a pool of photoreceptor-specific cDNAs originally created by subtractive hybridization of mRNAs from normal and photoreceptorless rd mouse retinas. Localization of the gene corresponding to mPNR to mouse Chr 9 near the rd7 locus made it a candidate for the site of the rd7 mutation. Northern analysis of total RNA isolated from rd7 mouse retinas revealed no detectable signal after hybridization with the mPNR cDNA probe. However, with reverse transcription-PCR, we were able to amplify different fragments of mPNR from rd7 retinal RNA and to sequence them directly. We found a 380-nt deletion in the coding region of the rd7 mPNR message that creates a frame shift and produces a premature stop codon. This deletion accounts for more than 32% of the normal protein and eliminates a portion of the DNA-binding domain. In addition, it may result in the rapid degradation of the rd7 mPNR message by the nonsense-mediated decay pathway, preventing the synthesis of the corresponding protein. Our findings demonstrate that mPNR expression is critical for the normal development and function of the photoreceptor cells. PMID- 10805812 TI - Coexistence of linear zones and pinwheels within orientation maps in cat visual cortex. AB - Revealing the layout of cortical maps is important both for understanding the processes involved in their development and for uncovering the mechanisms underlying neural computation. The typical organization of orientation maps in the cat visual cortex is radial; complete orientation cycles are mapped around orientation singularities. In contrast, long linear zones of orientation representation have been detected in the primary visual cortex of the tree shrew. In this study, we searched for the existence of long linear sequences and wide linear zones within orientation preference maps of the cat visual cortex. Optical imaging based on intrinsic signals was used. Long linear sequences and wide linear zones of preferred orientation were occasionally detected along the border between areas 17 and 18, as well as within area 18. Adjacent zones of distinct radial and linear organizations were observed across area 18 of a single hemisphere. However, radial and linear organizations were not necessarily segregated; long (7.5 mm) linear sequences of preferred orientation were found embedded within a typical pinwheel-like organization of orientation. We conclude that, although the radial organization is dominant, perfectly linear organization may develop and perform the processing related to orientation in the cat visual cortex. PMID- 10805813 TI - Accumulation of protease-resistant prion protein (PrP) and apoptosis of cerebellar granule cells in transgenic mice expressing a PrP insertional mutation. AB - We have generated lines of transgenic mice that express a mutant prion protein (PrP) containing 14 octapeptide repeats whose human homologue is associated with an inherited prion dementia. These mice develop a neurological illness with prominent ataxia at 65 or 240 days of age, depending on whether the transgene array is, respectively, homozygous or hemizygous. Starting from birth, mutant PrP is converted into a protease-resistant and detergent-insoluble form that resembles the scrapie isoform of PrP, and this form accumulates dramatically in many brain regions throughout the lifetime of the mice. As PrP accumulates, there is massive apoptosis of granule cells in the cerebellum. Our analysis provides important insights into the molecular pathogenesis of inherited prion disorders in humans. PMID- 10805814 TI - Why are angles misperceived? AB - Although it has long been apparent that observers tend to overestimate the magnitude of acute angles and underestimate obtuse ones, there is no consensus about why such distortions are seen. Geometrical modeling combined with psychophysical testing of human subjects indicates that these misperceptions are the result of an empirical strategy that resolves the inherent ambiguity of angular stimuli by generating percepts of the past significance of the stimulus rather than the geometry of its retinal projection. PMID- 10805815 TI - Acute decrease in net glutamate uptake during energy deprivation. AB - The extracellular glutamate concentration ([glu](o)) rises during cerebral ischemia, reaching levels capable of inducing delayed neuronal death. The mechanisms underlying this glutamate accumulation remain controversial. We used N methyl-D-aspartate receptors on CA3 pyramidal neurons as a real-time, on-site, glutamate sensor to identify the source of glutamate release in an in vitro model of ischemia. Using glutamate and L-trans-pyrrolidine-2,4-dicarboxylic acid (tPDC) as substrates and DL-threo-beta-benzyloxyaspartate (TBOA) as an inhibitor of glutamate transporters, we demonstrate that energy deprivation decreases net glutamate uptake within 2-3 min and later promotes reverse glutamate transport. This process accounts for up to 50% of the glutamate accumulation during energy deprivation. Enhanced action potential-independent vesicular release also contributes to the increase in [glu](o), by approximately 50%, but only once glutamate uptake is inhibited. These results indicate that a significant rise in [glu](o) already occurs during the first minutes of energy deprivation and is the consequence of reduced uptake and increased vesicular and nonvesicular release of glutamate. PMID- 10805817 TI - Postgerminative growth and lipid catabolism in oilseeds lacking the glyoxylate cycle. AB - The glyoxylate cycle is regarded as essential for postgerminative growth and seedling establishment in oilseed plants. We have identified two allelic Arabidopsis mutants, icl-1 and icl-2, which lack the glyoxylate cycle because of the absence of the key enzyme isocitrate lyase. These mutants demonstrate that the glyoxylate cycle is not essential for germination. Furthermore, photosynthesis can compensate for the absence of the glyoxylate cycle during postgerminative growth, and only when light intensity or day length is decreased does seedling establishment become compromised. The provision of exogenous sugars can overcome this growth deficiency. The icl mutants also demonstrate that the glyoxylate cycle is important for seedling survival and recovery after prolonged dark conditions that approximate growth in nature. Surprisingly, despite their inability to catalyze the net conversion of acetate to carbohydrate, mutant seedlings are able to break down storage lipids. Results suggest that lipids can be used as a source of carbon for respiration in germinating oilseeds and that products of fatty acid catabolism can pass from the peroxisome to the mitochondrion independently of the glyoxylate cycle. However, an additional anaplerotic source of carbon is required for lipid breakdown and seedling establishment. This source can be provided by the glyoxylate cycle or, in its absence, by exogenous sucrose or photosynthesis. PMID- 10805816 TI - Potent and nontoxic antisense oligonucleotides containing locked nucleic acids. AB - Insufficient efficacy and/or specificity of antisense oligonucleotides limit their in vivo usefulness. We demonstrate here that a high-affinity DNA analog, locked nucleic acid (LNA), confers several desired properties to antisense agents. Unlike DNA, LNA/DNA copolymers were not degraded readily in blood serum and cell extracts. However, like DNA, the LNA/DNA copolymers were capable of activating RNase H, an important antisense mechanism of action. In contrast to phosphorothioate-containing oligonucleotides, isosequential LNA analogs did not cause detectable toxic reactions in rat brain. LNA/DNA copolymers exhibited potent antisense activity on assay systems as disparate as a G-protein-coupled receptor in living rat brain and an Escherichia coli reporter gene. LNA containing oligonucleotides will likely be useful for many antisense applications. PMID- 10805818 TI - Nas-Walras equilibria of a large economy. AB - Individuals exchange contracts for the delivery of commodities in competitive markets and, simultaneously, act strategically; actions affect utilities across individuals directly or through the payoffs of contracts. This encompasses economies with asymmetric information. Nash-Walras equilibria exist for large economies, even if utility functions are not quasi-concave and choice sets are not convex, which is the case in standard settings; the separation of the purchase from the sale of contracts and the pooling of the deliveries on contracts guarantee that the markets for commodities clear. PMID- 10805822 TI - Prospective study of the association between sleep-disordered breathing and hypertension. AB - BACKGROUND: Sleep-disordered breathing is prevalent in the general population and has been linked to chronically elevated blood pressure in cross-sectional epidemiologic studies. We performed a prospective, population-based study of the association between objectively measured sleep-disordered breathing and hypertension (defined as a laboratory-measured blood pressure of at least 140/90 mm Hg or the use of antihypertensive medications). METHODS: We analyzed data on sleep-disordered breathing, blood pressure, habitus, and health history at base line and after four years of follow-up in 709 participants of the Wisconsin Sleep Cohort Study (and after eight years of follow-up in the case of 184 of these participants). Participants were assessed overnight by 18-channel polysomnography for sleep-disordered breathing, as defined by the apnea-hypopnea index (the number of episodes of apnea and hypopnea per hour of sleep). The odds ratios for the presence of hypertension at the four-year follow-up study according to the apnea-hypopnea index at base line were estimated after adjustment for base-line hypertension status, body-mass index, neck and waist circumference, age, sex, and weekly use of alcohol and cigarettes. RESULTS: Relative to the reference category of an apnea-hypopnea index of 0 events per hour at base line, the odds ratios for the presence of hypertension at follow-up were 1.42 (95 percent confidence interval, 1.13 to 1.78) with an apnea-hypopnea index of 0.1 to 4.9 events per hour at base line as compared with none, 2.03 (95 percent confidence interval, 1.29 to 3.17) with an apnea-hypopnea index of 5.0 to 14.9 events per hour, and 2.89 (95 percent confidence interval, 1.46 to 5.64) with an apnea-hypopnea index of 15.0 or more events per hour. CONCLUSIONS: We found a dose-response association between sleep-disordered breathing at base line and the presence of hypertension four years later that was independent of known confounding factors. The findings suggest that sleep-disordered breathing is likely to be a risk factor for hypertension and consequent cardiovascular morbidity in the general population. PMID- 10805823 TI - Effects of physiologic pacing versus ventricular pacing on the risk of stroke and death due to cardiovascular causes. Canadian Trial of Physiologic Pacing Investigators. AB - BACKGROUND: Evidence suggests that physiologic pacing (dual-chamber or atrial) may be superior to single-chamber (ventricular) pacing because it is associated with lower risks of atrial fibrillation, stroke, and death. These benefits have not been evaluated in a large, randomized, controlled trial. METHODS: At 32 Canadian centers, patients without chronic atrial fibrillation who were scheduled for a first implantation of a pacemaker to treat symptomatic bradycardia were eligible for enrollment. We randomly assigned patients to receive either a ventricular pacemaker or a physiologic pacemaker and followed them for an average of three years. The primary outcome was stroke or death due to cardiovascular causes. Secondary outcomes were death from any cause, atrial fibrillation, and hospitalization for heart failure. RESULTS: A total of 1474 patients were randomly assigned to receive a ventricular pacemaker and 1094 to receive a physiologic pacemaker. The annual rate of stroke or death due to cardiovascular causes was 5.5 percent with ventricular pacing, as compared with 4.9 percent with physiologic pacing (reduction in relative risk, 9.4 percent; 95 percent confidence interval, -10.5 to 25.7 percent [the negative value indicates an increase in risk]; P=0.33). The annual rate of atrial fibrillation was significantly lower among the patients in the physiologic-pacing group (5.3 percent) than among those in the ventricular-pacing group (6.6 percent), for a reduction in relative risk of 18.0 percent (95 percent confidence interval, 0.3 to 32.6 percent; P=0.05). The effect on the rate of atrial fibrillation was not apparent until two years after implantation. The observed annual rates of death from all causes and of hospitalization for heart failure were lower among the patients with a physiologic pacemaker than among those with a ventricular pacemaker, but not significantly so (annual rates of death, 6.6 percent with ventricular pacing and 6.3 percent with physiologic pacing; annual rates of hospitalization for heart failure, 3.5 percent and 3.1 percent, respectively). There were significantly more perioperative complications with physiologic pacing than with ventricular pacing (9.0 percent vs. 3.8 percent, P<0.001). CONCLUSIONS: Physiologic pacing provides little benefit over ventricular pacing for the prevention of stroke or death due to cardiovascular causes. PMID- 10805825 TI - Mortality from pneumonia in children in the United States, 1939 through 1996. AB - BACKGROUND AND METHODS: Pneumonia remains an important cause of childhood deaths throughout the world, but in developed countries, the mortality rate is decreasing. We reviewed death records for children in the United States from 1939 through 1996. A plot of the annual rates of change in the number of deaths from pneumonia was used to generate hypotheses about the influence of various events and interventions. We used data from the National Hospital Discharge Survey, the Medicaid program, and published reports to test these hypotheses. RESULTS: During the 58-year study period, the number of children who died from pneumonia declined by 97 percent, from 24,637 in 1939 to 800 in 1996. During the same period, the rate of mortality from other causes declined by 82 percent. There were steep declines in the mortality rates for pneumonia from 1944 to 1950, although the rate increased among older children in 1957, and there were sustained declines in all age groups from 1966 to 1982. From 1966 to 1982, the mortality declined by an average of 13.0 percent annually, and these decreases coincided with increases in the proportion of poor children covered by Medicaid, increases in rates of hospitalization for pneumonia, a narrowing of the gap between the mortality rate for black children and the rate for white children, and a convergence between the mortality rate in the South and the rates in the other three census regions. CONCLUSIONS: Since 1939, the rate of mortality from pneumonia in children in the United States has declined markedly. We hypothesize that the steep declines in the late 1940s are attributable to the use of penicillin, that the peak in 1957 was due to the influenza A pandemic, and that the sustained decline from 1966 through 1982 may be attributable in part to improved access to medical care for poor children. PMID- 10805824 TI - Beneficial effects of high dietary fiber intake in patients with type 2 diabetes mellitus. AB - BACKGROUND: The effect of increasing the intake of dietary fiber on glycemic control in patients with type 2 diabetes mellitus is controversial. METHODS: In a randomized, crossover study, we assigned 13 patients with type 2 diabetes mellitus to follow two diets, each for six weeks: a diet containing moderate amounts of fiber (total, 24 g; 8 g of soluble fiber and 16 g of insoluble fiber), as recommended by the American Diabetes Association (ADA), and a high-fiber diet (total, 50 g; 25 g of soluble fiber and 25 g of insoluble fiber), containing foods not fortified with fiber (unfortified foods). Both diets, prepared in a research kitchen, had the same macronutrient and energy content. We compared the effects of the two diets on glycemic control and plasma lipid concentrations. RESULTS: Compliance with the diets was excellent. During the sixth week, the high fiber diet, as compared with the the sixth week of the ADA diet, mean daily preprandial plasma glucose concentrations were 13 mg per deciliter [0.7 mmol per liter] lower (95 percent confidence interval, 1 to 24 mg per deciliter [0.1 to 1.3 mmol per liter]; P=0.04) and mean median difference, daily urinary glucose excretion 1.3 g (0.23; 95 percent confidence interval, 0.03 to 1.83 g; P= 0.008). The high-fiber diet also lowered the area under the curve for 24-hour plasma glucose and insulin concentrations, which were measured every two hours, by 10 percent (P=0.02) and 12 percent (P=0.05), respectively. The high-fiber diet reduced plasma total cholesterol concentrations by 6.7 percent (P=0.02), triglyceride concentrations by 10.2 percent (P=0.02), and very-low-density lipoprotein cholesterol concentrations by 12.5 percent (P=0.01). CONCLUSIONS: A high intake of dietary fiber, particularly of the soluble type, above the level recommended by the ADA, improves glycemic control, decreases hyperinsulinemia, and lowers plasma lipid concentrations in patients with type 2 diabetes. PMID- 10805826 TI - Images in clinical medicine. Upper-airway resistance syndrome. PMID- 10805827 TI - The effect of longevity on spending for acute and long-term care. AB - BACKGROUND: The proportion of the population made up of elderly persons in the United States is projected to increase from 13 percent of the population in 2000 to 20 percent by 2030. The implications for health care expenditures may be profound, because elderly persons use health care services at a greater rate than younger persons. We estimated total expenditures for acute and long-term care from the age of 65 years until death and in the last two years of life. METHODS: We combined data from Medicare, the National Mortality Followback Survey, and the National Medical Expenditure Survey to estimate total national expenditures for health care according to the age at death. We also simulated expenditures with the use of projected demographic characteristics of two cohorts: people turning 65 in 2000 and those turning 65 in 2015. RESULTS: Total expenditures (in 1996 dollars) from the age of 65 years until death increase substantially with longevity, from $31,181 for persons who die at the age of 65 years to more than $200,000 for those who die at the age of 90, in part because of steep increases in nursing home expenditures for very old persons. Spending in the last two years of life also increases with longevity, but a reduction in Medicare expenditures ($37,000 for persons who die at the age of 75 years and $21,000 for those who die at the age of 95) moderates the effect of the increase in nursing home expenditures ($6,000 for those who die at the age of 75 years and $32,000 for those who die at the age of 95). Health care spending for women is consistently higher than that for men, after adjustment for the increased longevity of women. Simulations show that increased longevity after the age of 65 years has a relatively small effect on the anticipated increase in spending, especially for services covered by Medicare, from 2000 to 2015. The effects of the larger number of people born in 1950 than in 1935 and the larger number of people surviving to the age of 65 years are much more important. CONCLUSIONS: In the United States, the effect of longevity on expenditures for acute care differs from its effect on expenditures for long-term care. Acute care expenditures, principally for hospital care and physicians' services, increase at a reduced rate as the age at death increases, whereas expenditures for long-term care increase at an accelerated rate. Increases in longevity after the age of 65 years may result in greater spending for long-term care, but the increase in the number of elderly persons has a more important effect on total spending. PMID- 10805829 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 14-2000. A 60-year-old farm worker with bilateral pneumonia. PMID- 10805828 TI - Prophylaxis against opportunistic infections in patients with human immunodeficiency virus infection. PMID- 10805830 TI - A New Feature -- This Week in the Journal. PMID- 10805831 TI - Dietary treatment of diabetes mellitus. PMID- 10805832 TI - Fortification of foods with folic acid--how much is enough? PMID- 10805833 TI - Correction: Case Records of the Massachusetts General Hospital (Case 1-2000). PMID- 10805834 TI - The impact of colorectal cancer screening on life expectancy. AB - METHODS: The purpose of this study was to assess the effect of screening for colorectal cancer on life expectancy and estimate the number of colonoscopies needed per life year saved. The declining exponential approximation of life expectancy was used to calculate the effect of colorectal cancer screening on expected remaining lifetime. The annual number of deaths from colorectal cancer and the size of the population were obtained from the vital statistics of the United States. Published reports were consulted to determine the decrease in mortality from colorectal cancer achieved by fecal occult blood testing, screening sigmoidoscopy or colonoscopy. A Markov chain analysis was used to determine the endoscopic resources required to screen and survey the entire population of U.S. residents age 50 years until death or age 85 years. RESULTS: Colorectal cancer decreases the life expectancy of U.S. residents aged 50 to 54 years by 292 days and those aged 70 to 74 years by 70 days. Screening with fecal occult blood tests extends expected lifetime of the 2 age groups by 51 and 12 days, respectively, whereas screening with sigmoidoscopy leads to increases of 86 and 21 days. Colonoscopic screening increases expected lifetime by 170 and 41 days, respectively. The number of colonoscopies needed to save 1 year of expected life ranges from 2.9 to 6.0, depending on the type of screening regimen used. CONCLUSIONS: The extension of life through screening colonoscopy is two or three times longer than the extension achieved through flexible sigmoidoscopy or fecal occult blood test, respectively. Although a large number of colonoscopies are required to screen the U.S. population, relatively few colonoscopies need to be invested per year of life expectancy saved. PMID- 10805835 TI - Screening colonoscopy in asymptomatic average-risk African Americans. AB - BACKGROUND: Recent data indicate that colorectal cancer incidence and mortality in white Americans have been declining since 1985 at a rate of 2% to 3% per year. In African Americans, however, mortality from colorectal cancer appears to be increasing. We sought to evaluate the prevalence of colonic neoplasia in asymptomatic African Americans. METHODS: We performed a cross-sectional colonoscopy screening study to determine the prevalence of colonic neoplasia in asymptomatic African Americans older than 50 years of age. RESULTS: One hundred sixty-six subjects were evaluated for the study of whom 121 (69 women) were deemed to be asymptomatic average-risk persons and completed colonoscopy. Forty two individuals (35%) had a total of 72 adenomas (67 tubular and 5 tubulovillous); 47 (65.3%) of these were proximal to the splenic flexure. Three subjects had an adenoma 1 cm or greater in diameter and none had severe dysplasia. CONCLUSIONS: The overall prevalence of adenomas in asymptomatic average-risk African Americans was comparable to that of previously described populations. The predominance of right-sided adenomas in this study confirms previous findings and is an area requiring further study. Until this issue is resolved, we suggest the use of colonoscopy rather than sigmoidoscopy for screening for colorectal neoplasia in asymptomatic, average-risk African Americans. PMID- 10805836 TI - Pain associated with phase III of the duodenal migrating motor complex in patients with postcholecystectomy biliary dyskinesia. AB - BACKGROUND: Correlation between various gastrointestinal events and particular aspects of the migrating motor complex has been reported. This study correlates postcholecystectomy pain to variations in biliary pressure associated with the duodenal motor cycle. METHODS: In 18 patients with postcholecystectomy pain and 10 control subjects, biliary and duodenal pressures were recorded simultaneously with microtransducers. After recording a spontaneous cycle, morphine was administered to induce a premature phase III and spasm of the sphincter of Oddi, and then cerulein was administered to stop the spasm. RESULTS: Transient but significant elevations of biliary pressure occurred at duodenal phase III in both groups, but a greater percentage of the patients developed pain during phase III (89% vs. 20%, p<0.01). Morphine produced premature phase III and biliary pressure elevation, which were accompanied by pain more frequently in the patients than in the control subjects (78% vs. 30%, p<0.05). Biliary pressure dropped after the cerulein injection, relieving the pain in 13 of 14 patients and in 2 of 3 control subjects who had morphine-induced pain. The phase III-related pain was relieved by endoscopic sphincterotomy in 14 of 15 patients. CONCLUSIONS: The cyclic elevation of biliary pressure in coordination with phase III of the duodenal motor cycle may contribute to the development of pain in patients with postcholecystectomy biliary dyskinesia. PMID- 10805837 TI - Grading ERCPs by degree of difficulty: a new concept to produce more meaningful outcome data. AB - BACKGROUND: Simple endoscopic retrograde cholangiopancreatography (ERCP) outcome measures such as success and complication rates may not allow direct comparisons among endoscopists or centers because procedure degree of difficulty can vary tremendously from case to case. We propose a new grading scale designed to objectively quantify ERCP degree of difficulty. METHODS: A 1 to 5 scale was devised to grade ERCPs according to their level of technical difficulty. A retrospective pilot study was performed to assess ERCP outcomes at our institution according to difficulty grade. The scale was then prospectively applied to all ERCPs during a 1-year period. RESULTS: In the pilot study, 209 of 231 (90%) ERCPs were technically successful, and 8 (3%) were followed by complications. Grade 1 to 4 procedures were more likely to succeed (94% vs. 74%, p< 0.05) and less likely to have associated complications (2% vs. 10%, p< 0.05) than grade 5/5B ERCPs. Of 187 ERCPs assessed prospectively, 166 (89%) were successful and 10 (5%) were followed by complications; 132 of 138 (96%) grade 1 to 4 procedures succeeded compared with 30 of 46 grade 5 to 5B ERCPs (65%, p<0.001), but complications were not significantly more frequent in grade 5 to 5B ERCPs (8.7% vs. 4.3%, p = not significant). CONCLUSIONS: Technical success was dependent on ERCP degree of difficulty, but complications were not. Outcome data that incorporate degree of difficulty information may be more meaningful, allowing endoscopist-to-endoscopist and center-to-center comparisons. PMID- 10805838 TI - Long-term outcome of endoscopic papillotomy for choledocholithiasis with cholecystolithiasis. AB - BACKGROUND: Although endoscopic papillotomy is now considered established treatment for choledocholithiasis, therapeutic results of endoscopic papillotomy alone without subsequent cholecystectomy in patients with cholecystolithiasis have not been well evaluated. The aim of this study was to assess the long-term outcome of endoscopic papillotomy for these patients. METHODS: Patients admitted with choledocholithiasis and cholecystolithiasis from 1976 to 1993 were studied retrospectively. Of 385 patients in whom the bile duct was cleared by endoscopic papillotomy and endoscopic stone extraction, 371 patients (195 men and 176 women; mean age 65.4 years) were followed. Predisposing risk factors for late complications were analyzed. RESULTS: The mean duration of follow-up was 7.7 years. Cholecystitis and recurrence of choledocholithiasis as late complications occurred in 22 cases (5.9%) and 36 cases (9.7%), respectively. Cholecystitis, including 1 severe case, resolved with conservative treatment. Recurrent choledocholithiasis was successfully treated endoscopically except in 1 case. No significant risk factors were identified for cholecystitis. The presence of pneumobilia (p = 0.0016) and the need for lithotripsy (p = 0.0342) were found to be significant risk factors for the recurrence of choledocholithiasis. CONCLUSIONS: Long-term outcome of endoscopic papillotomy in patients with choledocholithiasis and cholecystolithiasis was found to be relatively favorable. Cholecystectomy after endoscopic papillotomy is not always necessary in the management of cholecystolithiasis. PMID- 10805839 TI - A randomized, double-blind study of the use of droperidol for conscious sedation during therapeutic endoscopy in difficult to sedate patients. AB - BACKGROUND: Droperidol has been used in combination with narcotics and benzodiazepines to achieve conscious sedation. We performed a randomized, double blind, study of droperidol in patients at risk for difficult sedation scheduled for therapeutic endoscopy. METHODS: Patients with regular ethanol, narcotic, or benzodiazepine usage, suspected sphincter of Oddi dysfunction, or a history of difficult sedation were eligible for the study. Patients were randomized to receive either droperidol or placebo along with midazolam and meperidine as preprocedure sedation. Time to achieve sedation, interruptions due to undersedation, medication dosages, recovery time, and subjective assessments of sedation were recorded. RESULTS: One hundred one patients were randomized. The droperidol group had significantly fewer procedure interruptions and observer ratings of difficulty with sedation and required significantly less midazolam (23%) and meperidine (16%) than the placebo group. There were no significant differences in time to achieve sedation, incomplete procedures, procedure length, recovery room time, or complications. There were significantly higher observer ratings of the quality of sedation for patients who received droperidol. CONCLUSIONS: Droperidol is a useful adjunct to conscious sedation in patients who are difficult to sedate. Its use results in significantly fewer interruptions due to poor sedation and improved sedation ratings compared with sedation using midazolam and meperidine alone. PMID- 10805840 TI - Role of EUS in the preoperative localization of insulinomas compared with spiral CT. AB - BACKGROUND: Preoperative radiologic localization of insulinomas often fails because of the small size of these tumors. Endoscopic ultrasound (EUS) can localize insulinomas in up to 80% of the cases. The aim of this study was to compare EUS and computed tomography (CT) diagnostic accuracy for insulinomas. METHODS: We reviewed medical records from 12 patients (10 women) with a biochemical diagnosis of hypoglycemia and hyperinsulinism from 1 university hospital and 1 community hospital. A diagnosis of insulinoma was ultimately made in all cases and before surgery the patients underwent abdominal US, spiral CT and EUS in an attempt to precisely localize the tumor. Surgery was considered the standard for tumor localization. RESULTS: Ten tumors were benign (83.3%) and 2 were malignant (16.7%). The overall sensitivity of EUS in identifying insulinomas was 83.3% compared with 16.7% for CT. Tumors not detected by EUS had a mean size of 0.75 cm. EUS-guided fine-needle aspiration was possible in only 3 patients, with a positive cytologic diagnosis in 2 (66.6%). Tumors located in the head and body of the pancreas were identified by EUS in all patients, but those located in the tail were diagnosed in only 50% of the cases. CONCLUSIONS: EUS is superior to spiral CT and should replace it for the detection of pancreatic insulinomas. EUS identification depends on the site and size of the tumor. PMID- 10805841 TI - Nonmalignant obstruction is a common problem with metal stents in the treatment of esophageal cancer. AB - BACKGROUND: The use of metal stents for the treatment of dysphagia due to esophageal malignancy is an important advance because of ease of delivery and their self-expandable property. Obstruction due to tumor overgrowth is a recognized complication, but nonmalignant obstruction in patients with metal stents is rarely reported. METHODS: Database records of patients who had esophageal cancer and underwent metal stent insertion were reviewed. RESULTS: A total of 116 patients were seen between October 1993 and October 1997. Four types of metal stents had been used (Ultraflex, Z Stent, Wallstent, and Esophacoil). Detailed follow-up information was available for 81 patients, who constitute the study sample. Forty-nine (60%) stent obstructions were reported, 26 of the 49 (53%) were due to tumor overgrowth and 23 (47%) were not associated with malignancy. Histologic analysis of the nonmalignant obstructing tissue showed granulation tissue (56%), reactive hyperplasia (22%) and fibrosis (22%). CONCLUSIONS: Nonmalignant obstruction is a common although infrequently reported complication after placement of metal stents for esophageal cancer. The tissue response of the esophageal mucosa occurred with all 4 types of stents used. No specific characteristic of the stent or prior treatment seems to be related to obstruction of the stent in patients with either nonmalignant obstruction or tumor overgrowth. PMID- 10805842 TI - Methylene blue-directed biopsies improve detection of intestinal metaplasia and dysplasia in Barrett's esophagus. AB - BACKGROUND: Endoscopically applied methylene blue selectively stains specialized columnar epithelium in Barrett's esophagus. METHODS: The diagnostic yield and cost of cancer surveillance in patients with Barrett's esophagus using methylene blue-directed biopsies (MBDB) were compared with surveillance using a "jumbo" random biopsy technique in a prospective, sequential, controlled trial. Esophagogastroduodenoscopy was performed with either MBDB or random biopsy in a randomized sequence. The proportions of various types of epithelia in each biopsy were estimated and dysplasia was graded in a blinded fashion. RESULTS: Forty three patients with short- (n = 8), limited- (n = 10), and long-segment (n = 25) Barrett's esophagus were studied. Using MBDB technique, the average number of biopsies obtained per patient was significantly lower and the proportion of specialized columnar epithelium in each specimen was significantly higher compared with random biopsy. Dysplasia or cancer was diagnosed in significantly more MBDB specimens (12% vs. 6%, p = 0.004). Despite fewer biopsies per patient using MBDB, dysplasia or cancer was diagnosed in significantly more patients (44% vs. 28%, p = 0.03) than by random biopsy technique. MBDB cost less and detected more cancers than random biopsy. CONCLUSIONS: MBDB is a more accurate and cost effective technique than random biopsy for diagnosing specialized columnar epithelium and dysplasia/cancer, particularly in long-segment Barrett's esophagus. PMID- 10805843 TI - Endoscopic injection sclerotherapy for esophageal varices prolonged survival of patients with hepatocellular carcinoma complicating liver cirrhosis. AB - BACKGROUND: A prospective controlled study was performed between 1982 and 1991 to evaluate the efficacy of endoscopic injection sclerotherapy (EIS) in patients with esophageal varices complicated by hepatocellular carcinoma and liver cirrhosis. METHODS: The study included 83 patients with esophageal varices, hepatocellular carcinoma, and liver cirrhosis. Forty-three patients (group 1) underwent prophylactic EIS or emergent EIS for bleeding varices. EIS was performed weekly 4 to 6 times until the varices disappeared. The remaining 40 patients (group 2) underwent conservative therapy and did not undergo EIS. Survival rates were compared between the 2 groups. RESULTS: During the 5-year observational period, all patients who did not undergo EIS died. Sixteen in group 2 (40.0%) died of gastrointestinal bleeding including ruptured esophageal varices. In contrast, patients treated with EIS survived significantly longer (p<0.001). Nine patients (20.9%) treated with EIS experienced gastrointestinal bleeding as a result of which 5 (11.6%) died. EIS prolonged survival in patients classified as Child's A or B but did not affect survival in patients with Child's C hepatic function. EIS was effective in prolonging survival in patients with hepatocellular carcinomas smaller than 5 cm. However, EIS had no effect in patients with hepatocellular carcinomas that were larger than 5 cm. EIS prolonged survival only for patients with nodular hepatocellular carcinoma and had no effect in patients with massive and diffuse hepatocellular carcinoma. Further, EIS prolonged survival only for patients who did not have portal vein thrombosis. CONCLUSION: Based on this prospective study, we concluded that EIS was effective in prolonging the survival period of a select subset of patients with hepatocellular carcinoma. PMID- 10805844 TI - Absolute ethanol and 5% ethanolamine oleate are comparable for sclerotherapy of esophageal varices. AB - BACKGROUND: Endoscopic sclerotherapy is widely accepted as an effective treatment for the eradication of esophageal varices in patients with portal hypertension and a history of upper gastrointestinal bleeding. The objective of this study was to assess the effectiveness and safety of absolute ethanol as an alternative sclerosing agent to the commonly used 5% ethanolamine oleate. METHODS: One hundred fifty-seven patients with portal hypertension and a history of variceal bleeding were randomly assigned to sclerotherapy with absolute ethanol (n = 66) or 5% ethanolamine oleate (n = 91) between January 1992 and July 1994. Once eradication was achieved, these patients were prospectively followed until September 1998. RESULTS: Sclerotherapy with both sclerosants resulted in similar eradication rates (approximately 90%), with comparable numbers of sessions required for eradication (5.4 and 5.9 sessions for absolute ethanol and 5% ethanolamine oleate, respectively). Similar complication and recurrent bleeding rates were observed among both groups. CONCLUSION: Sclerotherapy with absolute ethanol is as effective as with 5% ethanolamine oleate in preventing further bleeding in patients with portal hypertension. PMID- 10805845 TI - Biliary cutaneous stent insertion via T-tube tract by choledochoscopy. AB - BACKGROUND: Postoperative biliary ductal dilation and stent insertion are mandatory for managing complicated residual hepatolithiasis with intrahepatic strictures. Usually this procedure is performed under fluoroscopic guidance. Choledochoscopic examination is a routine procedure for residual hepatolithiasis. This is the first report of choledochoscopic biliary cutaneous stent insertion. METHODS: Fifty-three patients underwent biliary cutaneous stent insertion under postoperative choledochoscopic guidance. Of the 53 patients, 35 had strictured intrahepatic ducts, 6 a stenotic hilum, 9 a tortuous fistula tract and 3 a choledochoduodenocutaneous fistula. RESULTS: A total of 120 procedures have been performed in the 53 patients. Two thirds necessitated a repeated procedure. No major complication was found except 2 patients experienced mild abdominal pain after the procedure. Neither fluoroscopy nor contrast medium was used. CONCLUSIONS: Choledochoscopic guidance is effective for postoperative biliary cutaneous stent insertion. It has the advantage of being technically easy and is relatively safe. PMID- 10805846 TI - Endoscopic laser therapy for palliation of patients with distal colorectal carcinoma: analysis of factors influencing long-term outcome. AB - BACKGROUND: Incurable rectal cancer often reduces the quality of life because of obstruction, bleeding, pain and tenesmus. For such symptoms palliative therapy is often carried out. METHODS: From 1986 to 1995, 219 patients, 118 men and 101 women, mean age 67 years, with distal colorectal adenocarcinoma were referred for palliative endoscopic laser therapy. In this retrospective analysis of outcome, patients were allocated to 3 subgroups according to their dominant symptom: obstruction, bleeding and others (soiling, diarrhea, tenesmus). After initial successful treatment, maintenance therapy was carried out in cases of obstruction at intervals of 2 to 4 months; patients with bleeding, tenesmus or diarrhea were retreated if there was recurrence of symptoms. RESULTS: Initial successful palliation was obtained in 198 patients (92%), with similar results in the 3 subgroups. Long-term, effective palliation was achieved in 160 patients (75%) of the total study population. Seventy-six patients (65.0%) with obstruction, 63 (82.9%) with bleeding and 21 (80.8%) with other symptoms remained symptom free until death or this analysis of results. There was a significantly negative relation between long-term successful outcome and local spread of the tumor in the obstruction (p = 0.040) and bleeding groups (p = 0.014). The total number of treatments was significantly higher if obstruction was present at presentation (p< 0.05) and if tumors were circumferential (p = 0.05). Major complications were perforation (4.1%), fistula (3.2%), abscess (1.7%) and bleeding (4.1%). There were 5 (possibly) procedure-related deaths. The survival rate was 44.4% at 1 year and 20.4% at 2 years. CONCLUSIONS: Laser therapy is effective for initial palliation but careful patient selection is necessary. Long-term palliation is less feasible in patients with obstruction. PMID- 10805847 TI - Endoscopic definitions of esophagogastric junction regional anatomy. PMID- 10805848 TI - Neuroendocrine carcinoma of the ampulla of vater. PMID- 10805849 TI - Hepato-biliary ascariasis. PMID- 10805850 TI - Posttransplantation lymphoproliferative disorder of the stomach. PMID- 10805851 TI - Stress ulcers of the stomach. PMID- 10805852 TI - Imaging of esophageal tumors with a water-filled condom and a catheter US probe. AB - BACKGROUND: High-frequency catheter ultrasound (C-EUS) probes provide high resolution images of the gastrointestinal tract. Their use in the esophagus is limited by the requirement for a water-filled lumen for good acoustical coupling. We have developed a system using a condom that provides a continuous column of water for high resolution C-EUS imaging of the esophagus. METHODS: Nine patients underwent condom C-EUS for evaluation of esophageal mucosal and submucosal tumors. A standard latex condom was attached to a 2-channel endoscope and filled with water after esophageal intubation. A 20 MHz C-EUS probe placed within the water-filled condom was used to characterize all lesions. RESULTS: The condom C EUS system provided a 360 degree, high resolution image throughout the length of the esophagus without air artifact and without the risk of aspiration associated with filling the esophageal lumen with water. Complete imaging of the tumor was obtained in all cases, and no additional C-EUS procedures were required. The condom C-EUS was subjectively rated superior overall in comparison to standard C EUS by 4 independent endosonographers. CONCLUSIONS: Condom C-EUS is a new method of high resolution imaging of the esophagus. This method provides a contained column of water within the esophagus that improves image quality and ease of use and may reduce the risk of aspiration. PMID- 10805853 TI - Colonoscopy technique with an external straightener. AB - BACKGROUND: An external straightener for colonoscopy which enables proper compression of the abdomen during the entire examination has been developed. METHODS: Beginning January 1, 1997, patients undergoing outpatient colonoscopy were subjected to either manual abdominal compression or compression with an external straightener. Two hundred patients were divided into two groups of 100 patients each: group A (colonoscopy using traditional methods; 38 men, 62 women, mean age 59.6 years, range 18 to 80) and group B (colonoscopy with the help of the external straightener; 40 men, 60 women, mean age 59.8 years, range 16 to 75). Fifteen minutes after the examination, each patient completed a form that assessed the degree of pain during the procedure (no pain, mild, moderate, severe). RESULTS: The ileocecal valve was reached in 89 cases in group A and 94 cases in group B. The average time required to reach the valve was 9.34+/-4 minutes (range 4 to 25) in group A and 6.97+/-3.37 minutes (range 2 to 21) in group B (p<0.001). With regard to the degree of pain, the results for groups A and B were, respectively: no pain = 25% and 40%, mild = 29% and 34%, moderate = 30% and 20%, severe = 16% and 6% (p< 0.001). CONCLUSIONS: The external straightener reduced examination time and decreased the degree of patient pain compared with traditional methods. PMID- 10805854 TI - Pancreatic duct obstruction caused by malignant islet cell tumors of the pancreas. PMID- 10805855 TI - Dysphagia due to transmural migration of surgical material into the esophagus nine years after Nissen fundoplication. PMID- 10805856 TI - Acute esophageal necrosis associated with gastric volvulus. PMID- 10805857 TI - Successful endoscopic management of a perforated gastric dysplastic lesion after endoscopic mucosal resection. PMID- 10805858 TI - Endoscopic laser treatment of obstructing polypropylene mesh after vertical banded gastroplasty. PMID- 10805859 TI - The "salpinx" polyp: a unique presentation of Crohn's disease. PMID- 10805860 TI - Solitary hamartomatous polyp of Peutz-Jeghers type in the jejunum resected endoscopically. PMID- 10805861 TI - Lozenge-induced esophagitis. PMID- 10805862 TI - Screening colonoscopy: option or preference? PMID- 10805863 TI - Live endoscopy demonstrations are great, but... PMID- 10805864 TI - The beginning: endoscopic sphincterotomy PMID- 10805865 TI - Back to the future: the first papillotomy at Erlangen. PMID- 10805866 TI - In the beginning. "Report of endoscopical papillotomy". PMID- 10805867 TI - Reminiscences of some current experts and early American sphincterotomy. PMID- 10805868 TI - Is the technique of endoscopic sclerotherapy and ligation (ESL) optimum? PMID- 10805869 TI - Endoscopic lasso: is it new? PMID- 10805870 TI - Endoscopic lasso: is it new? PMID- 10805871 TI - "Lasso" technique for retrieval of impacted biliary stent. PMID- 10805872 TI - Amebic colitis simulating ulcerative colitis. PMID- 10805873 TI - Presidential address: challenge to our specialty: the vascular surgeon in the year 2010. PMID- 10805874 TI - Surgical repair of aneurysms involving the suprarenal, visceral, and lower thoracic aortic segments: early results and late outcome. AB - OBJECTIVE: The purpose of this study is to review our experience with surgical repair of lower thoracoabdominal and suprarenal aortic aneurysms to determine early and late survival rates and identify factors influencing morbidity and survival among these patients. MATERIALS: From 1989 through 1998, 165 consecutive patients underwent repair of 108 thoracoabdominal (55 group III and 53 group IV) and 57 suprarenal aneurysms. The study group consisted of 109 men and 56 women with a mean age of 70 years (median, 70 years; range, 29-89 years). Mean aneurysm diameter was 6.9 cm (median, 6.5 cm; range, 4-12 cm). There were 125 aneurysms (76%) repaired electively; 40 repairs (24%) were nonelective. The cause of 12 aneurysms (7%) was chronic aortic dissection; the remaining 153 (93%) were degenerative aneurysms. RESULTS: The early postoperative (30-day) mortality rates were 7% (9/125) for elective and 23% (9/40) for nonelective operations (P =.016). For both elective and urgent procedures, early mortality was 1.8% (1/57) for suprarenal aneurysm repair, 11% (6/53) for group IV thoracoabdominal aneurysms, and 20% (11/55) for group III thoracoabdominal aneurysms (P =.013, suprarenal vs group III). Spinal cord ischemia occurred after 6% (10/165) of aneurysm repairs (4% paraplegia, 2% paraparesis). None of the 57 suprarenal aneurysm repairs were complicated by spinal cord ischemia, whereas it occurred in 2% (1/53) of group IV thoracoabdominal aneurysms and 16% (9/55) of group III thoracoabdominal aneurysms (P =.001, suprarenal vs group III; P =. 016, group IV vs group III). Three (25%) of the 12 patients with dissection developed spinal cord ischemia; this compared with seven (5%) of 153 patients with degenerative aneurysms (P =.027). The cumulative 3-year survival rate for the entire series was 71% (95% CI, 64%-79%), and 5-year survival was 50% (95% CI, 40%-60%). CONCLUSIONS: Aneurysms involving the suprarenal, visceral, and lower thoracic aorta may be repaired with acceptable perioperative mortality and late survival rates. The risk of spinal cord ischemia is increased for patients with aortic dissection and may be stratified according to the proximal extent of the aneurysm. PMID- 10805875 TI - Incidence of femoral and popliteal artery aneurysms in patients with abdominal aortic aneurysms. AB - OBJECTIVE: The purpose of this study was to determine the incidence of femoral and popliteal aneurysms in men and women who have abdominal aortic aneurysms (AAAs) and to assess potential etiologic differences in patients with and without these lower extremity aneurysms. METHODS: We studied 313 consecutive patients with AAAs encountered from 1995 to 1998 who underwent prospective ultrasound scanning to detect the presence or absence of femoral and popliteal aneurysms. Patients with and without these extremity aneurysms were compared for differences in potential etiologic risk factors with each other and with a statewide population of patients with AAAs. RESULTS: A total of 51 femoral and popliteal aneurysms were encountered, all occurring in male patients. Among the 251 men with AAAs, the incidence of femoral or popliteal aneurysms was 14%, compared with 0% among the 62 women with AAAs (P <.01). A family history of aneurysmal disease was present in only one (3%) of the 36 men with these extremity arterial aneurysms, a significant finding (P <.01) when compared with the family history that was positive for aneurysmal disease in 14 women (23%). Peripheral arterial occlusive disease affected 14 (39%) of the 36 men with peripheral arterial aneurysms versus 20 (9%) of the 215 men without these aneurysms (P <.01). Most other etiologic variables studied proved not to be different among the various groups of patients examined. CONCLUSION: The incidence of femoral and popliteal aneurysms in persons with AAAs appears higher than that noted previously. Femoral and popliteal aneurysmal disease preferentially affects men; however, the basis for this sex difference is unknown. Few common etiologic factors differed between men with and without these extremity aneurysms. Most femoral and popliteal artery aneurysms in this study were undetectable on physical examination, suggesting that ultrasound scanning is appropriate in the recognition of peripheral aneurysms among men with AAAs. PMID- 10805876 TI - Factors affecting the diagnosis of peripheral vascular disease before vascular surgery referral. AB - OBJECTIVE: Many new patients evaluated by vascular surgeons are referred by internal medicine physicians (IMPs). Objectives shared by vascular surgeons and IMPs include early identification of peripheral arterial disease (PAD), improved referral relationships, and reduction of health care costs. The approach to PAD by IMPs and identification of deficiencies that might contribute to suboptimal care form the basis for this report. METHODS: An anonymous survey was mailed to all IMPs (n = 843) in the central and southern parts of Illinois. Questions concerned IMP demographics, approach to diagnostic testing, referral patterns, perception of adequacy of education of PAD, and how often parts of the history and physical examination for PAD would be performed on the initial office visit of a hypothetical 65-year-old male with hypertension (each answer measured as 0% 25%, 25%-50%, 50%-75%, and 75%-100% of the time completed). RESULTS: There was a response from 360 IMPs: 230 IMPs (27.3%) returned the questionnaire, and 130 IMPs (15.4%) declined to participate. Practice locations for IMPs returning the questionnaire included rural (36%), suburban (22%), and urban (40%). Practice types included academic (7%), solo private (29%), group private (53%), and other (14%). A history of cardiac disease was obtained most of the time by 92% of IMPs (75%-100% answer category). Histories for pulmonary disease, diabetes mellitus, stroke, and smoking were obtained most of the time with similar frequencies (85%, 86%, 73%, and 96%, respectively). In contrast, only 37% obtained a history for claudication, and 26% obtained a history for foot ulceration 75% to 100% of the time (P <.05, all comparisons). Examination of the heart (95%) and lungs (96%) occurred most of the time (75%-100% answer category) compared with each part of the pulse examination (range, 34%-60%; P <.05, all comparisons) and aortic aneurysm palpation (39%; P <.05). If pedal pulses were absent, examination by IMPs with Doppler scan and ankle-arm indices were mostly distributed in the 0% to 25% answer category (79% and 79%, respectively). After suspecting PAD, most IMPs obtained diagnostic tests first compared with specialist referral: carotid disease (91% vs 9%), aortic aneurysm (91% vs 9%), and lower extremity PAD (86% vs 14%). Initial referral patterns were made to vascular surgeons (49%), general surgeons (33%), cardiothoracic surgeons (13%), cardiologists (4%), and radiologists (1%). Most IMPs believed medical school (70%) and residency (73%) provided adequate training for PAD diagnosis. CONCLUSIONS: Deficiencies may exist in the identification of PAD by IMPs that could adversely affect diagnosis, time to referral, health care costs, and ultimately, patient outcome. Improvements in medical school education and IMP training in the diagnosis of PAD are needed. PMID- 10805877 TI - Clinical failure after percutaneous transluminal angioplasty of the superficial femoral and popliteal arteries. AB - OBJECTIVE: Anatomic patency after percutaneous transluminal angioplasty (PTA) of the superficial femoral and popliteal arteries does not guarantee clinical success. The aim of this report is to determine the causes of clinical failure after PTA. METHODS: The records of all patients who have undergone PTA of the femoropopliteal arterial segment by our vascular group were retrospectively reviewed. Only patients with complete records and at least one postprocedure clinical and anatomic assessment within the same 30-day time interval were included. Success was defined according to the Society for Vascular Surgery/International Society for Cardiovascular Surgery Ad Hoc Subcommittee on Reporting Standards for Endovascular Procedures. Anatomic cumulative patency and clinical success were calculated according to life table analysis on an intent-to treat basis. RESULTS: We identified 85 patients who met inclusion criteria. We treated 112 lesions with an average stenosis of 80% +/- 16% and lesion length of 2.3 +/- 1.8 cm. Technical failure occurred in six (5.4%) of 112 lesions. Cumulative clinical success was 69% at 1 year, 54% at 2 years, 49% at 3 years, and 40% at 4 years. Anatomic patency was 74% at 1 year, 62% at 2 years, 57% at 3 years, and 52% at 4 years. There were 45 clinical failures; of these, twenty seven (60%) occurred in conjunction with anatomic failure. Anatomic failure was due to restenosis in 12 patients (44%), occlusion in eight patients (30%), and restenosis with progression of disease in six patients (22%). Anatomic failure at the time of the procedure occurred in one patient (4%). Clinical failure occurred despite anatomic patency in the remaining 18 patients (40%). Etiology for clinical failure in this latter group included progression of disease within the treated vessel in 12 patients (67%), iliac disease in three patients (17%), tibial disease in two patients (11%), and bypass graft failure in one patient (5%). Fifty percent of all 45 clinical failures were successfully treated with supplemental percutaneous procedures. CONCLUSION: A PTA is an acceptable therapeutic option for the treatment of focal occlusive disease of the femoropopliteal arterial segment. Most clinical failures were due to anatomic failure, but a significant number occurred despite patency at the PTA site. Although primary clinical success rates were inferior to surgical bypass graft, supplemental PTA was possible in 50% of patients. Repeat percutaneous treatment may extend the interval of clinical success and may obviate the need for surgical bypass graft. PMID- 10805878 TI - Comparing patency rates between external iliac and common iliac artery stents. AB - PURPOSE: Placement of intraluminal stents in the common iliac artery (CIA) and external iliac artery (EIA) has become an accepted therapy for treating localized arterial stenoses. The purpose of this study was to compare anatomic patency rates of stents placed in the EIA and CIA for occlusive disease. METHODS: A radiologic computer database was used to identify 69 consecutive male patients at the Minneapolis Veterans Affairs Medical Center from February 1, 1993, through January 31, 1999, who underwent placement of 98 stents (82 Wallstents and 16 Palmaz stents) for physiologically significant iliac artery occlusive disease and varying degrees of chronic limb ischemia. Patients were followed up with surveillance duplex ultrasound scanning examinations 1 day after procedure, 3 months after procedure, and then at 6-month intervals after stent placement. Follow-up angiograms were performed for patients with duplex ultrasound scans that revealed velocities greater than 300 cm/s. Patient risk factors, iliac artery runoff, concomitant outflow procedures, and anatomic patency rates were compared between patients receiving EIA stents and those receiving CIA stents. RESULTS: The mean age for the EIA stent group was 69 +/- 1 years versus 66 +/- 1 years (P =.03) for the CIA stent group. Mean follow-up was 21.4 +/- 2.1 months (+/- SE) for all patients. Patients with EIA stents had more ischemic lower limbs when compared with patients who had CIA stents (P =.05). No differences were found between groups in risk factor analysis (P = not significant). Lesion lengths were similar between groups: EIA, 4.6 +/- 0.6 cm, and CIA, 5. 3 +/- 0.8 cm (P = not significant). The following differences were noted on primary patency rates (EIA group vs CIA group): 1 year (93% vs 88%), 2 years (91% vs 85%), and 3 years (90% vs 78%) (Cox proportional hazards; P =.13). CONCLUSIONS: Anatomic patency rates for EIA and CIA stents appear to be similar despite the fact that patients with EIA stents were older and had more ischemic limbs compared with the patients who had CIA stents. PMID- 10805879 TI - The anatomy of deep venous thrombosis of the lower extremity. AB - BACKGROUND: The diagnosis, treatment, and long-term sequelae of lower extremity deep venous thrombosis (DVT) depend on the anatomic location and extent of the process, yet a lack of such fundamental knowledge has limited the development of effective protocols for managing patients with DVT. METHODS: Venograms with evidence of acute DVT were evaluated, and the extent of the thrombotic process was recorded and correlated with the clinical presentation. Thrombi were classified according to the venous segments involved and to the thrombus' isolation to one segment or multiple segments. The left-to-right ratio of the DVT was assessed for various etiologic subgroups. RESULTS: Among 2762 venograms performed in 2541 patients over a 10-year period, there were 885 cases (34.8%) of DVT documented. Of these cases, 344 cases (39%) were idiopathic, 307 cases (35%) were postoperative, 84 cases (10%) occurred in the setting of malignancy, and 70 cases (8%) occurred as the result of trauma. Distal thrombi were more common than proximal thrombi, with calf involvement in 734 patients (83%), femoropopliteal involvement in 470 patients (53%), and iliac involvement in 75 patients (9%). The most common site of thrombus was the peroneal vein, which was involved in 595 patients (67%). The ratio of left-to-right-sided DVT was 1.32:1 overall but was greater for proximal thrombi, with a ratio of 2.4:1 for iliac DVT versus 1.3:1 for infrainguinal DVT. The preponderance of left-sided DVT appeared to be related to the high-frequency, left common iliac vein involvement; the left-to-right ratio was much closer to equality (1.09:1) for isolated infrainguinal DVT. The anatomic configuration of the DVT was correlated with the etiologic subgroup; postoperative DVTs were more often distal, whereas DVT developing in the setting of malignancy was more frequently proximal and often right sided. Proximal, left sided DVTs were common in the idiopathic subgroup, presumably as a result of undiagnosed left iliac vein webs. CONCLUSIONS: The frequency of distal vein involvement greatly exceeds that of proximal involvement in patients with DVTs. Proximal DVTs are more frequently left sided, whereas distal DVTs occur with a more equal left-to-right distribution. The anatomic extent of DVTs appears to depend on the etiology of the process. These observations may shed light on the pathophysiology of venous thrombosis. The findings are of value in planning therapeutic interventions directed at venous recanalization. PMID- 10805880 TI - Carotid endarterectomy: characterization of recent increases in procedure rates. AB - INTRODUCTION: Recent increases in the rate of carotid endarterectomies (CEAs) have been attributed to results of clinical trials demonstrating efficacy when CEA is performed in centers of excellence. Subsequent population-based data suggest that trial results may not be matched in the community. This study was undertaken to characterize trends in CEA procedure rates after the dissemination of trial data and to describe any change in patient outcomes with population based data from a single state. METHODS: Hospital administrative data on CEAs from 1992 to 1996 (n = 45,744) were obtained for the state of Florida. Annualized CEA rates per 100, 000 Florida residents were analyzed to determine trends in patient age, sex, admission type, size of hospital beds, ownership type and teaching status, and annual hospital and surgeon CEA volume. Outcomes were examined to track trends in complication rates. RESULTS: The annual number of CEA procedures increased 74% from 63.7 per 100,000 residents per year to 110.8 per 100,000 residents per year between 1992 and 1996. A single large increase occurred during the second half of 1994 when CEAs increased 73.5% from 16.6 per 100, 000 residents per quarter to 28.8 per 100,000 residents per quarter after a clinical alert on benefits to CEAs in asymptomatic patients. Over 5 years, there were significant trends toward more nonemergent admissions, and more procedures were performed in high-volume hospitals and by high-volume surgeons. Procedure rates in both women and very elderly patients increased more than 70%, which was in step with younger patients and men. The incidence of inpatient stroke and death declined over the 5-year period, whereas the rate of perioperative myocardial infarction remained constant. CONCLUSIONS: Experience from Florida indicates that CEA rates increased as results of the Asymptomatic Carotid Artery Study disseminated. Trial results have been broadly interpreted to include women and very elderly patients. More patients are being referred to busier hospitals and to high-volume surgeons, which should continue to result in better patient outcomes. PMID- 10805881 TI - Effective hemodynamic diameter: an intrinsic property of vein grafts with predictive value for patency. AB - INTRODUCTION: Conduit size and quality are major determinants of the long-term success of infrainguinal autologous vein grafting. However, accurate measurement of the internal diameter of vein grafts is difficult given their variable wall thickness and taper. The purpose of this study was to define the "effective" internal diameter of a vein graft according to its hemodynamic properties and to determine its significance for graft patency. METHODS: Sixty infrainguinal bypass grafts performed on 57 patients were evaluated intraoperatively. Proximal and distal graft pressure and blood flow (Q(meas)) were measured with fluid-filled catheter transduction and ultrasonic transit-time flowimetry, respectively, after unclamping. Waveforms were recorded digitally at 200 Hz under baseline conditions and after stimulation with 60 mg of papaverine. According to Fourier transformation of the measured pressure gradient (DeltaP), the Womersley solution for fluid flow in a straight rigid tube was used to calculate theoretical flow waveforms (Q(calc)) for a range of graft diameters. The theoretical waveforms were then compared with the measured flow waveforms and the best-fit diameter chosen as the "effective hemodynamic diameter" (EHD). Only grafts in which the correlation coefficient of Q(calc) versus Q(meas) was more than 0.90 were accepted (n = 47) to assure validity of the hemodynamic model. After a mean follow-up of 12.5 months (range, 0.1-43.9 months), patency was determined by the life table method. Hemodynamic and clinical variables were tabulated, and their effect on patency determined the use of univariate and multivariate Cox regression. RESULTS: Mean EHD was 4.1 +/- 0.1 mm with a range of 2.5 to 5.7 mm. Administration of papaverine caused profound changes in DeltaP (+78% +/- 17%) and Q(meas) (+71% +/- 12%) as expected, but had no effect on EHD (+0.05% +/- 0.1%). Univariate regression identified five variables associated with decreased secondary patency (P <.10): low EHD, conduit source other than the greater saphenous vein, high baseline DeltaP(mean), female sex, and redo operation. Of these, only low EHD was significant after multivariate analysis (P =.03). Patency of small diameter grafts (EHD < 3.6 mm; n = 11) was compared with patency of larger grafts (EHD > 3.6 mm; n = 36) to test a frequently espoused clinical guideline. Grafts with an EHD less than 3.6 mm exhibited significantly lower secondary patency compared with larger grafts (P =.0001). The positive and negative predictive values for an EHD less than 3.6 mm for secondary graft failure for grafts with at least 1 year follow-up were 86% and 88%, respectively. CONCLUSION: An EHD is a unique parameter that quantifies conduit size and has a significant impact on vein graft patency. An EHD less than 3.6 mm portends graft failure. PMID- 10805882 TI - Improving the outcomes of carotid endarterectomy: results of a statewide quality improvement project. AB - OBJECTIVE: The purpose of this study was to establish the statewide outcomes for carotid endarterectomy (CEA) and to facilitate improvement in outcomes through feedback, peer discussion, and ongoing process and outcome measurement. METHODS: The Medicare Part A claims files were used to identify all Medicare patients undergoing CEA in Iowa during two 12-month time periods (January 1994-December 1994 and June 1995-May 1996). Medical record abstraction was used to obtain surgical indications, perioperative care process, and outcome information. Confidential reports were provided to each hospital (N = 30) where the procedure was performed. Surgeons performing the procedure (N = 79) were invited to meetings to discuss care process variation and outcomes. Voluntary participation was solicited in a standardized program of ongoing hospital-based data collection of CEA process and outcome data. RESULTS: The statewide combined stroke or mortality rate decreased from 7.8% in 1994 to 4.0% in the 1995 to 1996 time period (P <.001). Fourteen hospitals, accounting for 74% of the statewide cases, participated in ongoing data collection. The combined stroke or mortality rate in these hospitals decreased significantly (P <.05) over time from 6.5% (1994) to 3.7% (1995-1996) to 1.8% (June 1997-May 1998). The use of intraoperative assessment of the operative site (20% in 1994, 46% in 1997-1998) and patch angioplasty (14% in 1994, 30% in 1997-1998) increased significantly during this time in the participating hospitals. CONCLUSIONS: Confidential feedback of outcome and process data for CEA may lead to change in perioperative care processes and improved outcomes. Standardized community-based outcome analysis should become routine for CEA to ensure that optimum results are being achieved. PMID- 10805883 TI - Carotid endarterectomy in octogenarians: comparison with patient characteristics and outcomes in younger patients. AB - OBJECTIVES: Carotid endarterectomy has been shown to be of clear benefit to selected patients. However, recent trials of carotid endarterectomy versus best medical therapy have excluded octogenarians, and some authors have suggested that carotid endarterectomy would have an unfavorable cost-benefit relationship in octogenarians. We compared patients and results for carotid endarterectomy in octogenarians and younger patients. METHODS: We reviewed the results for 582 primary carotid endarterectomies (90 in octogenarians and 492 in younger patients) performed in 528 patients between February 1, 1985, and January 31, 1998 (all data were collected prospectively for the most recent 301 carotid endarterectomies). Conventional surgical technique was used with general anesthesia, selective shunting, and selective patching. Main outcome measures were perioperative and late ipsilateral stroke and death. RESULTS: The two groups were similar with respect to indications for carotid endarterectomy and patient characteristics, except that octogenarians were more likely to have histories of congestive heart failure or hypertension and less likely to have histories of smoking or chronic lung disease. Carotid endarterectomy was performed for asymptomatic disease in 27% of the octogenarians and 33% of the younger patients (P =.31). Stenosis was >/=80% in 90% of the octogenarians and 78% of the younger patients (P =.014). Perioperative strokes, all of which were ipsilateral, occurred in one octogenarian (1.1%) and eight younger patients (1.6%, P = 1.00). No octogenarians and two younger patients died within 30 days of surgery (P = 1.00). Length of stay and direct costs associated with carotid endarterectomy were similar for octogenarians and younger patients. Late strokes occurred in two octogenarians (one ipsilateral) and four younger patients (two ipsilateral). Life table estimates of freedom from ipsilateral stroke at 2 years were 98% and 97% for octogenarians and younger patients, respectively (log-rank P =.69), and life table estimates of patient survival at 4 years were 81% and 89% for octogenarians and younger patients, respectively (P =.11). Octogenarians represented an increasing fraction of the carotid endarterectomies performed during the study period. CONCLUSIONS: Octogenarians selected for carotid endarterectomy were similar to younger patients with respect to indications for carotid endarterectomy and comorbidities. Early mortality, early and late neurologic outcome, complications, and resource utilization were similar for the two groups, and more than 75% of octogenarians survived 4 years after undergoing carotid endarterectomy. Cost-benefit analyses for carotid endarterectomy, which are highly sensitive to expected patient survival, might not be pertinent to individual patient situations. Intellectually intact octogenarians without unusually severe comorbidities are good candidates for and should be offered the benefits of carotid endarterectomy. PMID- 10805884 TI - Mitogenicity and release of vascular endothelial growth factor with and without heparin from fibrin glue. AB - PURPOSE: Fibrin glue (FG) has been used for local cytokine delivery on both vascular grafts and angioplasty sites. We measured the diffusive release of vascular endothelial growth factor (VEGF) and heparin from FG and the mitogenic activity of VEGF with and without heparin in FG on canine endothelial cells (ECs) and smooth muscle cells (SMCs). METHODS: Release of VEGF labeled with iodine 125 and tritiated heparin from FG into the overlying media was serially measured over 96 hours, and the data are reported as the mean percent released +/- SD. Proliferation assays measuring tritiated thymidine incorporation were performed for ECs and SMCs plated in media with 10% serum on FG containing various concentrations of VEGF and heparin. Media was placed on the FG for 24 hours and removed before plating cells to minimize the effect of the released, soluble VEGF and heparin. RESULTS: At 24 hours, 54% +/- 1% and 58% +/- 1% of the radioactive VEGF and heparin were released, respectively, with minimal release thereafter (58% +/- 1% and 66% +/- 1% at 96 hours). The ECs, SMCs, or media only (no cells) was plated on FG containing radioactive VEGF in an immediate or 24-hour delayed fashion for 72 hours to determine the percent release of VEGF into the media with the two different methods of plating. Cell type and the presence or absence of cells did not affect VEGF release, but there was three times more VEGF in the media for the immediate versus delayed plating (P <.001). Without heparin, VEGF at 100 ng/mL or more in the FG was needed to induce EC proliferation. Heparin at 5 U/mL enhanced EC proliferation at the VEGF dose of 100 ng/mL as compared wtih no heparin (P <.001), but not at the VEGF dose of 1000 ng/mL, which likely represents a maximal response. With heparin at 500 U/mL, the ECs died. In contrast, VEGF, in the presence or absence of heparin, did not affect SMC proliferation. CONCLUSIONS: We conclude that FG with VEGF at 1000 ng/mL and heparin at 5 U/mL is the optimal concentration for in vivo use because this may encourage EC, but not SMC, proliferation. The VEGF at 1000 ng/mL should leave mitogenic concentrations of VEGF intact after the initial, diffusive loss, and the addition of heparin at 5 U/mL may enhance VEGF mitogenic activity. PMID- 10805885 TI - Phosphorus 31 nuclear magnetic resonance spectroscopy suggests a mitochondrial defect in claudicating skeletal muscle. AB - OBJECTIVE: Decreased oxygen supply is generally accepted as the primary cause of muscle dysfunction in patients with peripheral arterial occlusive disease (PAOD) and intermittent claudication, although reported morphologic changes in the mitochondria of claudicating muscle suggest that impaired energy utilization may also play a role. With the measurement of the phosphate-rich compounds of muscle energy metabolism (adenosinetriphosphate [ATP], adenosinediphosphate [ADP], and phosphocreatine [PCr]) and pH, phosphorus P 31 magnetic resonance spectroscopy ((31)P MRS) provides a unique, noninvasive method to investigate this hypothesis further. METHODS: Calf muscle bioenergetics were studied in 12 men with moderate claudication (ankle-brachial index >/=0.5 and .5, Pearson moment correlation). CONCLUSIONS: Phosphorus 31 MRS provides the first direct evidence of defective energy metabolism in the mitochondria of claudicating calf muscle. This defect appears to be independent of both arterial flow and the severity of occlusive disease in patients with mild to moderate claudication. Coupled with documented ultrastructural and DNA abnormalities in the mitochondria of claudicating skeletal muscle, these data provide evidence for a secondary cause of muscle dysfunction in intermittent claudication. PMID- 10805886 TI - The migratory response to platelet-derived growth factor of smooth muscle cells isolated from synthetic vascular grafts in a canine model. AB - OBJECTIVE: Previous studies on smooth muscle cells (SMCs) harvested from implanted synthetic grafts demonstrate increased production of platelet-derived growth factor (PDGF) but decreased proliferative response compared with aortic SMCs. The purpose of this study was to determine the migratory response of graft versus aortic SMCs. METHODS: Thoracoabdominal grafts were implanted in beagles. The SMCs were harvested from the graft and infrarenal aorta. Migration was determined with the use of a razor-scrape assay and computerized image analysis. RESULTS: The mean distance migrated and the number of cells that migrated were greater in graft SMCs at baseline (185 +/- 18 micrometer and 108 +/- 17 cells) compared with aortic cells (110 +/- 10 micrometer and 42 +/- 5 cells)(P <.05). Baseline differences persisted after treatment with antibodies to PDGF. The addition of PDGF (10 ng/mL) resulted in increased migration in both graft (229 +/ 23 micrometer and 146 +/- 20 cells) and aortic SMCs (130 +/- 9 micrometer and 70 +/- 5 cells) compared with baseline (P <.05). The relative increase in response to PDGF was similar between the two groups (P = not significant). CONCLUSIONS: Graft SMCs differ phenotypically from aortic SMCs; they exhibit increased basal migration that is independent of autocrine stimulation by PDGF. In contrast to their blunted proliferative response, graft SMCs have a similar migratory response to PDGF compared with aortic SMCs. PMID- 10805887 TI - Aneurysm rupture after endovascular repair using the AneuRx stent graft. AB - OBJECTIVE: The purpose of this study was to determine the cause and frequency of aneurysm rupture after endovascular aneurysm repair. METHODS: We reviewed each patient who sustained aneurysm rupture among all patients enrolled for endovascular aortic aneurysm repair in phases I, II, and III of the US AneuRx Multicenter Clinical Trial from June 1996 through October 1999. RESULTS: A total of 1067 patients were enrolled for endovascular aneurysm repair. The AneuRx stent graft was successfully implanted in 1046 patients (98%). Endovascular repair was unsuccessful in 21 patients (2%); 13 patients (1%) were converted to open aneurysm repair. Among these, two patients (0.2%) sustained aneurysm rupture due to procedure-related instrumentation and underwent open surgical conversion. Aneurysm rupture has occurred in seven patients (0.7%) 3 weeks to 24 months (mean, 16 months) after successful endovascular repair. Four patients survived open surgical repair, and three patients died within 30 days. Overall rupture related mortality was 0.5% and included late deaths after rupture. Before rupture, two patients had endoleak and aneurysm enlargement, and five patients had no endoleak and no aneurysm enlargement. After aneurysm rupture all seven patients had evidence suggesting that there was poor fixation of the stent graft at the proximal distal, or iliac junction fixation sites. The two patients with endoleak declined recommended open surgical or endovascular repair, which could have prevented aneurysm rupture. In retrospect, the five patients without endoleak could potentially have avoided rupture with better patient selection, better stent graft positioning, or reinforcement of fixation points with stent graft extenders. The probability of no aneurysm rupture for all patients undergoing endovascular repair is 0.996 +/- 0.002 at 1 year and 0.974 +/- 0.011 at 2 years by life table analysis with the longest follow-up of 41 months. CONCLUSION: The early risk of aneurysm rupture after endovascular aneurysm repair is low. However, the possibility of rupture persists even in patients with no endoleak after the procedure. Therefore, all patients treated with endovascular aneurysm repair should continue to be monitored after the procedure. Patients with evidence suggesting insecure stent graft fixation should undergo further endovascular treatment or open surgical repair. PMID- 10805888 TI - Guidelines for perioperative cardiac evaluation from the American College of Cardiology/American Heart Association task force are effective for stratifying cardiac risk before aortic surgery. AB - PURPOSE: We assessed whether the American College of Cardiology/American Heart Association (ACC/AHA) task force guidelines for perioperative cardiac evaluation could reliably stratify cardiac risk before aortic surgery. METHODS: We retrospectively applied the guidelines to a closed database, set up prospectively. The setting was a referral center in an institutional practice with hospitalized patients. The closed database included 133 patients who had a routine cardiac examination, which comprised an estimation of functional capacity and noninvasive testing, before aortic surgery. This cardiac evaluation led to the proposal of coronarography in 23 patients and to treating an underlying coronary artery disease in 21 patients (including three myocardial revascularizations). One patient died after myocardial revascularization, and two patients died of cardiac causes after aortic surgery. The algorithm of the ACC/AHA guidelines was applied independently by two investigators to each patient's file that was included in the existing database. The main outcome measure was a comparison between cardiac risk stratification with the ACC/AHA guidelines and the results of the routine cardiac evaluation. RESULTS: The ACC/AHA guidelines were successfully applied to all 133 files by the two investigators. After applying the algorithm, 73 patients were stratified as low cardiac risk, and 60 patients were stratified as high risk. The 21 patients who had undergone a preoperative coronary artery disease optimization were stratified as high risk by means of the ACC/AHA guidelines. The patients who died from cardiac causes were stratified as high risk by means of the ACC/AHA guidelines, whereas none of the patients stratified as low risk died during hospitalization. CONCLUSION: The ACC/AHA guidelines were effective in stratifying cardiac risk by using clinical predictors and an estimate of the physical capacity of the patient. Their use may allow a reduction in unnecessary noninvasive testing in patients stratified as being at low risk, while permitting the selection of all patients likely to benefit from preoperative coronary artery disease optimization. PMID- 10805889 TI - Surgical treatment of 50 carotid dissections: indications and results. AB - PURPOSE: This article analyzes the course of 48 patients with 49 chronic carotid dissections (who were treated surgically at our institution after a median anticoagulation period of 9 months because of a persistent high-grade stenosis or an aneurysm) and the course of one additional patient with acute carotid dissection (who underwent early operative reconstruction 12 hours after onset because of fluctuating neurologic symptoms). METHODS: All medical and surgical records and imaging studies were reviewed retrospectively. All histologic specimens were reevaluated by a single pathologist to assess the cause of dissection. Follow-up of 41 patients (85%) after 70 months (range, 1-190 months) consisted of an examination of the extracranial vessels in the neck by Doppler ultrasound scanning and a questionnaire about the patients' medical history and their personal appraisals of cranial nerve function. RESULTS: Seventy percent of the dissections had developed spontaneously; 18% were caused by trauma; 12% of all patients (22% of the women) had a fibromuscular dysplasia. Indication for surgery was a high-grade persisting stenosis and a persisting or newly developed aneurysm. Flow restoration was achieved by resection and vein graft replacement in 40 cases (80%) and thromboendarterectomy and patch angioplasty in three cases (6%). Gradual dilatation was performed and effective in two cases (4%). Five internal carotid arteries (10%) had to be clipped because dissection extended into the skull base. One patient died of intracranial bleeding. Five patients (10%) experienced the development of a recurrent minor stroke (ipsilateral, 4 patients; contralateral, 1 patient). Cranial nerve damage could not be avoided in 29 cases (58%) but were transient in most of the cases. During follow-up, one patient died of unrelated reasons, and only one patient had experienced the development of a neurologic event of unknown cause. CONCLUSION: Chronic carotid dissection can be effectively treated by surgical reconstruction to prevent further ischemic or thromboembolic complications, if medical treatment for 6 months with anticoagulation failed or if carotid aneurysms and/or high-grade carotid stenosis persisted or have newly developed. PMID- 10805890 TI - The contribution of the external carotid artery to cerebral perfusion in carotid disease. AB - PURPOSE: In the presence of carotid occlusion, the external carotid artery (ECA) becomes an important source of cerebral blood flow, especially if the circle of Willis is incomplete. The contribution of the ECA to hemispheric blood flow in patients with severe ipsilateral carotid stenosis has never been previously investigated. METHODS: One hundred eight patients were monitored during sequential cross-clamping of the external (ECA) and then ipsilateral internal carotid artery (ICA) during carotid endarterectomy using transcranial Doppler sonography (TCD) (Neuroguard CDS, Los Angeles, Calif), to measure middle cerebral artery blood flow velocity, and near-infrared spectroscopy, to measure regional cerebral oxygen saturation (CsO(2)) (Invos 3100A; Somanetics, Troy, Mich). RESULTS: On the ipsilateral ECA cross-clamp, the median fall in CsO(2) was 3% (interquartile range, 1%-4%; P <.0001). On addition of the ICA cross-clamp there was a further fall of 3% and a total fall of 6% (3%-9%; P <.0001). The median percentage fall in middle cerebral artery blood flow velocity on ECA clamping was 12% (4%-24%; P <.0001); on ICA clamping it was 48% (25%-74%; P <.0001). Falls in TCD on ECA clamping were greater with increasing severity of ipsilateral ICA stenosis. The correlation between CsO(2) and TCD on external clamping, although less strong than that on internal clamping, was statistically significant r = 0.32; P =.01; Spearman rank correlation). CONCLUSIONS: The falls in TCD and CsO(2) were of a similar order of magnitude and must therefore reflect a fall in cerebral perfusion. The ipsilateral ECA contributes significantly to intracranial blood flow and oxygen saturation in severe carotid stenosis. PMID- 10805891 TI - Cellular repopulation of human vein allograft bypass grafts. AB - PURPOSE: Vein allografts are an alternative conduit for patients lacking available autogenous vein. The ability to develop a neoendothelium is a potential advantage of allografts over other nonautogenous grafts, because endothelial cells have been shown to play numerous essential roles in vessel survival. However, repopulation by endothelial cells has not previously been demonstrated or characterized in human subjects. METHODS: In our prospective trial, 40 patients (20 men, 20 women) underwent cryo- preserved saphenous vein bypass grafting procedures for limb salvage. Several patients underwent multiple grafting procedures. All grafts were sampled at implantation. During the 31 month follow-up interval, 22 allografts were explanted at the time of revision or subsequent surgical procedure. All grafts (22 of 22) demonstrated intact endothelium at implantation and explantation. Seventeen explantation biopsy samples (seven from men, 10 from women) from 16 patients (seven men, nine women) were adequate for further histologic and immunofluorescent analysis. Explants were stained with hematoxylin-eosin and immunohistochemical markers to quantitate rejection and also underwent fluorescence in-situ hybridization, with probes for X and Y chromosomes and counterstain for nuclear envelope. Cells were counted as XX, XY, XO, YO, or unstained. The endothelium and vessel walls were analyzed for origin of cells based on sex-mismatched transplants, with sex-matched transplants serving as controls. RESULTS: Evidence of cellular damage was noted in all explanted allografts, and moderate or severe rejection (lymphocyte infiltrate, +CD3, +CD8, +CR3, cytotoxic granules) was noted in six explanted allografts (29%). All allografts demonstrated intact endothelium (complete or partial), at the time of both implantation and explantation. Sex-matched (male to male) control explants showed only male cells, as expected. Male donor-female recipient transplants showed complete repopulation by recipient (female) cells in nine of 10 cases (90%), whereas one case (10%) demonstrated partial repopulation (a mosaic of male and female cells). One patient's slides were unreadable. Findings in cells of the allograft wall were identical to those of the endothelium (nine recipient-only cells and one mosaic). Complete absence of donor cells was noted as early as 1 week after implantation, but mosaicism was demonstrated in one patient 3 months after grafting. No relationship could be demonstrated between repopulation and time (P >.05), quantity of rejection (P >.05), or donor age (P >.05). CONCLUSION: Both the endothelial lining and vessel wall of venous allografts repopulate with cells of recipient origin, resulting in either a completely novel cellular constituency or a mosaic of host and donor cells. The loss of donor cells may be mediated by apoptosis or rejection, and the rate of migration of repopulating host cells is, at this point, unclear. Although the development of a completely endothelial-lined conduit offers a potential advantage over other alternative conduits, the functional status of the neoendothelium and repopulated vessel wall and their role in maintenance of allograft patency require further investigation. PMID- 10805892 TI - Intestinal permeability is increased in patients with intermittent claudication. AB - PURPOSE: Intermittent claudication can be regarded as repeated ischemia reperfusion injury, which can induce a generalized increase in vascular permeability, including in intestine. The lactulose mannitol test (L/M test) was performed in patients with intermittent claudication to evaluate the change in intestinal permeability when they were forced to walk. METHODS: The L/M test was performed in 11 patients with intermittent claudication and 11 control subjects without intermittent claudication. The test was performed at rest and after exercise. Patients walked on a treadmill until they stopped because of pain. The control subjects were forced to walk as far as 200 m on a treadmill. The L/M test was repeated in the patients after successful arterial reconstruction. Then the patients were instructed to walk the same distance at the same speed as they had before surgery. RESULTS: In patients, the mean L/M ratio after exercise was significantly higher than the mean L/M ratio when they were at rest (0.068 +/- 0.053 vs 0.022 +/- 0.009, P <.05). In control subjects, however, no significant difference was observed between the mean L/M ratios after exercise and at rest. The mean L/M ratio after exercise, decreased from 0.068 +/- 0.053 to 0.018 +/- 0. 016 after successful arterial reconstruction (P <.05) in patients. CONCLUSION: Intestinal permeability, determined by means of the L/M test, significantly increased in patients with intermittent claudication after exercise, and it diminished after arterial reconstruction. The L/M test is, consequently, a new noninvasive method to reflect ischemia in lower limbs during exercise. PMID- 10805893 TI - Interposition vein cuff anastomosis alters wall shear stress distribution in the recipient artery. AB - OBJECTIVE: Interposition of a vein cuff between a prosthetic infrainguinal bypass graft and a recipient infrageniculate artery can improve graft patency. There is evidence that the improved performance may be explained by a redistribution of myointimal hyperplasia (MIH) away from the critical areas at the heel and toe of the cuff-artery anastomosis. It is widely accepted that there is an association between hemodynamic forces, more specifically, low wall shear stress (WSS), and the development of MIH. The aim of this study was to determine whether the reported redistribution of MIH in the interposition vein cuff (IVC) may be explained by differences in magnitude and distribution of WSS. Design of Study and Method: Detailed flow velocity measurements were made in life-size models of conventional end-to-side (ETS) and IVC anastomoses using a two-component laser Doppler anemometer under pulsatile flow conditions. Velocity vectors were determined in the plane of symmetry of the anastomosis, and the variation of WSS was estimated from near-wall velocity measurements on the floor and upper wall of the artery. RESULTS: The main flow features in the ETS anastomosis were flow separation at the graft hood, strong radial velocity at the heel, and a stagnation point on the floor of the artery that moved slightly during the flow cycle. In the IVC anastomosis, a coherent vortex that occupied most of the cuff volume was present from the systolic deceleration phase to end diastole. A stagnation point on the anastomosis floor was found to oscillate by about 4 mm. Critical regions of low mean WSS (ie, below 0.5 N/m(2)) were identified. In the ETS anastomosis, they were found at the heel and along the floor. In the IVC anastomosis, low mean WSS was found only on the floor, and it was generally less extensive than in the ETS anastomosis. CONCLUSION: The vein cuff anastomosis alters the mean WSS distribution within the recipient artery and removes the area of low WSS at the heel. This may explain the redistribution of MIH away from important sites in the recipient artery. PMID- 10805894 TI - Changes in the cooling rate and medium improve the vascular function in cryopreserved porcine femoral arteries. AB - PURPOSE: The purpose of this study was to design an adequate technique with which to cryopreserve pig femoral arteries and to assess the influence of storage times in vascular function. METHODS: Fifty-two femoral arteries were distributed in seven groups. In group A (control), 10 arteries were studied after harvest; in groups B1 and B2, 19 arteries were suspended in RPMI 1640 plus fetal calf serum plus dimethylsulfoxide and were cryopreserved at 1 degrees C per minute or 0.3 degrees C per minute, respectively. In groups C1 to C4, 23 arteries were suspended in modified Krebs-Henseleit plus dimethylsulfoxide plus sucrose, cryopreserved at 0.7 degrees C per minute, and kept frozen for 1, 15, 60, or 180 days, respectively. After being thawed, arteries were examined for contraction and endothelial-dependent vasodilation (organ bath studies), antithrombotic properties of the endothelial layer(perfusion studies), and vessel structure (electron microscopy). RESULTS: Endothelial cells were present in both cryopreserved and control arteries. The control vessels showed a mean contraction to norepinephrine (10(-7) mol/L) of 13010 +/- 3181 mg. Arteries in groups B1 and B2 did not respond to norepinephrine. Contraction in groups C1 to C4 was as follows: C1, 5354 +/- 1222 mg; C2, 5187 +/- 2672 mg; C3, 6867 +/- 2292 mg; C4, 7000 +/- 2858 mg, which represent 50% of the control values (P <.001). Vasodilation was similar in control (99% +/- 3%) and cryopreserved arteries (C1, 90% +/- 13%; C2, 93% +/- 12%; C3, 89% +/- 15%; C4, 88% +/- 22%). Storage time did not influence vascular function. Platelet interaction was almost absent and similar in all groups. CONCLUSION: A modified cryopreservation technique preserves endothelial function independently of the storage time up to 6 months. PMID- 10805895 TI - Association of the 4G/5G polymorphism in the promoter region of plasminogen activator inhibitor-1 with abdominal aortic aneurysms. AB - PURPOSE: A familial component has previously been identified in 11% to 20% of patients with abdominal aortic aneurysms (AAAs). The genetic basis of familial AAA remains elusive, however. Matrix metalloproteinases have been implicated in aneurysm development; and plasmin, a serine protease, activates metalloproteinases. Plasminogen activator inhibitor-1 (PAI-1) regulates plasmin activation through the tissue plasminogen activators. A polymorphism within the promoter area of PAI-1 has been described that modifies PAI-1 expression and consequently plasminogen activation. The 4G homozygous variant is associated with increased PAI-1 expression and consequently reduced plasmin activity and therefore may be selected against in-familial AAA. The purpose of this study was to investigate the incidence of the 4G/5G insertion/deletion polymorphism in the promoter area of the PAI-1 gene in a population with AAA. METHODS: Patients seen at a tertiary referral center for repair of abdominal aortic aneurysms were recruited. DNA was extracted from blood. Primers were designed to amplify a 99 (5G)-base pair (bp) and a 98 (4G)-bp fragment bracketing the polymorphism. The 5' primer was mutated to allow a restriction endonuclease to cleave the 5G polymorphism into a 77-bp and a 22-bp fragment. Samples were run on agarose gels and stained with ethidium bromide. RESULTS: One hundred ninety patients with AAAs, including 39 patients with strong family histories and 163 controls were examined. The frequency of the 4G:5G alleles in the AAA population and in the control population was 0.6:0.4. However, 26% of patients with familial AAA were homozygous 5G compared with 13% of the control population. The 4G-allele frequency was 0.47 in the familial AAAs, compared with 0.62 in the nonfamilial patients (P =.02) and 0.61 in the control population (P =.03). CONCLUSION: The selection against the 4G4G genotype in the familial AAA population may indicate a role for PAI in the development of AAA in this population. PMID- 10805896 TI - Immediate postoperative thrombolytic therapy: an aggressive strategy for neurologic salvage when cerebral thromboembolism complicates carotid endarterectomy. AB - A 42-year-old man with a high-grade left internal carotid artery (ICA) stenosis demonstrated on a duplex scan was referred to us. A cerebral arteriogram confirmed a greater than 90% left internal carotid stenosis, but with the unexpected finding of a moderate amount of thrombus in the proximal ICA. He underwent emergent left carotid endarterectomy, but during the operation, only a small amount of thrombus was identified as adherent to the atherosclerotic plaque. he awakened in the operating room with a dense right hemiplegia and aphasia. Immediate reexploration demonstrated a patent endarterectomy site, a distal thromboembolectomy was performed without extraction of thrombus, and urokinase (250,000 Units) was infused into the distal ICA. He reawakened with an unchanged right hemiplegia and aphasia. The patient then underwent an urgent postoperative carotid and cerebral arteriogram that demonstrated an embolus to the middle cerebral artery. he was treated with the superselective infusion of urokinase (500,000 Units), with almost complete resolution of the clot. Over the course of the next 48 hours, the patient made a nearly complete neurologic recovery, and he was discharged from the hospital with only a slight facial droop. At 2 months' follow-up he was completely neurologically healthy. To our knowledge this is the first reported case of urokinase administered in the immediate postoperative period in the angiography suite to treat a thromboembolus complicating a carotid endarterectomy. PMID- 10805897 TI - Use of superficial femoral vein for hemodialysis arteriovenous access. AB - Maintaining hemodialysis access in the expanding number of patients with end stage renal disease is a difficult and challenging problem. Published guidelines outline the initial recommendations for hemodialysis access; however, there is little consensus about the most appropriate options for the subset of patients with repeated access failures and/or unsuitable veins. Two case reports are presented describing the use of composite saphenous-superficial femoral vein autogenous accesses placed in the upper and lower extremities. The function of the autogenous accesses appeared to be similar to a mature arteriovenous fistula in the short-term, although further longitudinal studies are required. The superficial femoral vein may be a useful hemodialysis access conduit for patients with limited access options. PMID- 10805899 TI - Intravenous leiomyomatosis with cardiac extension: tumor thrombectomy through an abdominal approach. AB - Intravenous leiomyomatosis is an uncommon vascular tumor that may be seen with potentially life-threatening symptoms resulting from intracardiac extension. This tumor is frequently misdiagnosed and treated without appropriate preoperative imaging and planning, which at times leads to inadequate treatment and incomplete resections. The appropriate therapy is complete excision of the tumor. We describe a patient who was treated with a new approach involving a single-stage operation without the need for median sternotomy, cardiopulmonary bypass graft, or hypothermic arrest by resection of the tumor from the point of attachment in the abdominal portion of the inferior vena cava. PMID- 10805898 TI - A surgically treated case of Takayasu's arteritis complicated by aortic dissections localized in the ascending and abdominal aortae. AB - Because complication by aortic dissection is markedly rare in patients with Takayasu's arteritis, a limited number of reports have been published regarding surgically treated cases of Takayasu's arteritis that is complicated by aortic dissection. When graft replacement of the ascending aorta and aortic arch and extra-anatomic bypass grafting were performed in a 72-year-old japanese woman with Takayasu's arteritis, which was complicated by aortic dissections localized in the ascending and abdominal aortae, the postoperative course of this patient was satisfactory. It was considered that the media became friable in this patient because of the presence of Takayasu's arteritis and that hypertension that persisted for a long time caused the independent development of aortic dissections in the ascending and abdominal aortae. PMID- 10805900 TI - Localized dissection and delayed rupture of the abdominal aorta after extracorporeal shock wave lithotripsy. AB - Extracorporeal shock wave lithotripsy (ESWL) represents the preferred treatment for most upper ureteric and renal calculi. Complication rates associated with ESWL are low, justifying the enthusiasm and acceptance of this treatment modality. As the technique has become more widely available, some deleterious effects on the kidneys and the surrounding tissues are increasingly recognized. We report on the rupture of a severely calcified abdominal aorta in a 65-year-old man who underwent 3 months of ESWL treatment earlier for renal calculi. The patient was seen with an acute recrudescence of a long-standing abdominal and left flank pain, which began immediately after the last of the three sessions of ESWL and was associated with an episode of hypotension that occurred an hour before admission. Patient history and chronologic course of events strongly suggest the role of ESWL in the genesis of abdominal aorta rupture. PMID- 10805901 TI - Aortic pseudoaneurysm secondary to pancreatitis. AB - A patient with a pseudoaneurysm of the distal abdominal aorta, which developed as the result of complicated gallstone pancreatitis, is reported. The aorta was bypassed with an axillofemoral-femorofemoral graft; the pseudoaneurysm was resected, and the infrarenal aorta was oversewn. The patient survived a complicated postoperative course and was discharged with well-perfused lower extremities. No previous description of a pseudoaneurysm of the aorta as a complication of pancreatitis was found on review of the literature. PMID- 10805902 TI - The Vascular Surgery Sub-Board: progress report. PMID- 10805903 TI - Potential problems with industry-supported clinical research. PMID- 10805904 TI - Regarding "Popliteal entrapment: more common than previously recognized". PMID- 10805905 TI - Reply PMID- 10805907 TI - Reply PMID- 10805906 TI - The use of endovascular stents in the treatment of penetrating ulcers of the thoracic aorta. PMID- 10805909 TI - Reply PMID- 10805908 TI - Photoplethysmography and calf muscle function after subfascial endoscopic perforator ligation. PMID- 10805910 TI - Two-dimensional vibrational correlation spectroscopy of in vitro hydroxyapatite maturation. AB - Two-dimensional (2-D) Raman and 2-D IR correlation spectroscopy are applied to analyze changes in the nu(4) region of the IR spectrum and in the nu(1) region of the Raman spectrum during the maturation of hydroxyapatite (HA) following the solution-mediated conversion of amorphous calcium phosphate (ACP) to HA. The nu(1) region of the Raman spectrum exhibits a frequency shift and sharpening during the maturation. Comparison of the experimental and simulated 2-D plots for this process suggests that the shift of a single peak, rather than a change in the relative intensity of two overlapped bands, is responsible for the observed spectral changes. The nu(4) mode of the PO(3-)(4) ion (T(2) symmetry in the free species) splits into a triplet with components near 563, 575, and 603 cm(-1) in HA. In addition, broad features appear at 540 and 617 cm(-1). During the latest stages of the maturation, an OH(-) librational mode develops at approximately 632 cm(-1). Changes in the relative intensities of three components of the nu(4) mode are not all correlated with each other. The synchronous 2-D plots reveal that the 563 and 603 cm(-1) pair are positively correlated while the feature at 575 cm(-1) is absent. A 587 cm(-1) mode arising from ACP is negatively correlated with the 563 and 603 cm(-1) pair and is both synchronously (positively) and asynchronously correlated with the 540 cm(-1) feature during the early stages of the maturation but is absent from 2-D plots of the later stages of the maturation. Cross correlations between the nu(4) mode and the nu(1),nu(3) contour generally confirm and extend previous assignments for the latter spectral region. Finally, the suitability of the 2-D approach for analysis of IR spectral images is examined through studies of HA crystallinity in a human iliac crest biopsy sample. Trabecular bone contains a fraction of HA that is more crystalline and mature than could be achieved in vitro during the room temperature ACP --> HA interconversion. PMID- 10805911 TI - Vibrational spectroscopic characterization of new calcium phosphate bioactive coatings. AB - In this work calcium phosphate (CaP) compounds with different PO(3-)(4)/HPO(2 )(4) R molar ratios in the 0.65-149 range were synthesized. In fact, all these CaPs contain different amounts of HPO(2-)(4) and PO(3-)(4) ions as well as the amorphous precursors (tricalcium phosphate and octacalcium phosphate) of hydroxyapatite deposition, which was shown by in vitro and in vivo measurements. Spectroscopical IR and Raman results showed the presence of bands whose intensity ratio can be related to the molar ratio R; in particular, the Raman I(962)/I(987) and the IR I(1035)/I(1125) intensity ratios were characterized as markers of the molar ratio. For these CaP compounds a nucleation model, which was based on the ability of HPO(2-)(4) ions to form strong H bonds with PO(3-)(4) ions, was proposed. PMID- 10805912 TI - Binding of copper(II) to carnosine: Raman and IR spectroscopic study. AB - A comparative Raman and FTIR study of carnosine, a dipeptide present in several mammalian tissues, and its complexes with copper(II) at different pH values was carried out. The neutral imidazole ring gives rise to some bands that appear at different wavenumbers, depending on whether the imidazole ring is in the tautomeric form II or I. At pH 7 and 9 the molecule exists in equilibrium between the two tautomeric forms; tautomer I is predominant. Metal coordination is a factor that affects the tautomeric equilibrium, and the copper(II) coordination site can be monitored by using some Raman marker bands such as the vC(4)=C(5) band. On the basis of the vibrational results, conclusions can be drawn on the functional groups involved in the Cu(II) chelation and on the species existing in the Cu(II)-carnosine system. At neutral and basic pH the most relevant species formed when the Cu(II)/carnosine molar ratio is not very different from unity is a dimer, [Cu(2)L(2)H(-2)](0). In this complex the ligand coordinates the metal via the N (amino), O (carboxylate), and N (amide) donor atoms while the N(tau) nitrogen atoms of the imidazole rings (tautomer II) bridge the copper(II) ions. At a slightly acidic pH the two monomeric complexes [CuLH](2+) and [CuL](+) were present. In the former the imidazole ring takes part in the Cu(II) coordination in the tautomeric I form whereas in the latter it is protonated and not bound to Cu(II). PMID- 10805913 TI - Pharmacologic application of fourier transform IR spectroscopy: in vivo toxicity of carbon tetrachloride on rat liver. AB - Microsomal fractions from rat liver were examined by means of Fourier transform IR (FTIR) spectroscopy to study the in vivo toxic effect of carbon tetrachloride administered by intraperitoneal injection. Lipid content was significantly enhanced in the liver of treated rats compared with untreated ones. The level of saturated fatty acids largely increased while that of unsaturated acids slightly decreased as a consequence of lipid peroxidation induced by the xenobiotic compound. The conformational structure of membrane proteins was changed, which was shown by the large decrease in the alpha-helical configuration. In the polysaccharide region we observed an important loss in glucidic structures that could be related to the metabolic changes caused by carbon tetrachloride intoxication. Thus, FTIR spectroscopy appears to be a useful tool to rapidly investigate the chemical alterations induced by this drug in liver microsomes and to correlate them with biochemical and physiological data. PMID- 10805914 TI - Effects of pH and chloride concentration on resonance Raman spectra of human myeloperoxidase and Raman microspectroscopic analysis of enzyme state in azurophilic granules. AB - Resonance Raman spectra of human myeloperoxidase were examined at pH 3.3-10.5 in the absence and presence of chloride ions. Among the porphyrin vibrational bands, the core-size marker bands showed particularly large wavenumber downshifts on going from pH 8.7 to 5.3 with a transition midpoint at pH 6.5 in the absence of chloride ions. The chloride ions did not affect the spectrum at a pH below 5.3 and above 8.7 whereas an increase of chloride concentration at neutral pH caused spectral changes similar to those observed upon pH lowering. Analogous effects were also observed on the Raman intensity. In addition, the stretching mode of the bond between the heme Fe and proximal histidine shifted by -2 cm(-1) on going from pH 8.7 to 5.3. Decomposition of the nu(3) band revealed the presence of two components, which was confirmed by an isosbestic point in the absorption spectra. The observed spectral changes indicated the existence of alkaline and acidic forms of the enzyme. The pK of interconversion was 6.5, and it was increased by binding of chloride ions. The porphyrin core was slightly expanded in the acidic form compared to that in the alkaline form. A molecular mechanism of the porphyrin core expansion was proposed on the basis of the X-ray crystal structure. The pH-spectrum relationships obtained for the isolated enzyme were applied to in situ analysis of the state of myeloperoxidase in azurophilic granules of living neutrophils. The enzyme was stored in the acidic form and kept inactive in catalyzing HOCl production. PMID- 10805915 TI - In situ Raman microspectroscopic identification and localization of carotenoids: approach to monitoring of UV-B irradiation stress on Antarctic fungus. AB - The in situ Raman microspectroscopic properties of an Antarctic fungus are investigated to assess the nature and the spatial localization of the main chromophores and to study their spectral changes under enhanced UV-B irradiation. The Raman spectroscopic features of spores in situ are consistent with those of carotenoid-like pigments. In particular, the Raman shifts seem to be related either to the frequency modes of long conjugated double-bond carotenoids or to protein bound beta-carotene. The spectroscopic analysis at different spore depths clearly shows the strongest Raman signal arises from cell wall and membrane structures. The intensity of such a signal shows a drastic reduction upon UV-B irradiation without any significant frequency change. The use of Raman microspectroscopy for nondestructively monitoring the UV-B effects on Arthrobotrys ferox spores is also discussed. PMID- 10805916 TI - Vibrational analysis and spectra of orotic acid. AB - The IR and Raman spectra of polycrystalline anhydrous orotic acid and its N1, N3, and O12 trideuterated isotopomer are recorded in the 4000-40 cm(-1) spectral interval as part of a series of vibrational analyses of nucleosides, nucleotides, and related compounds carried out in our laboratory. The frequencies of the fundamental transitions and the potential energy distributions of the 39 normal modes of orotic acid and its isotopomer are calculated by an ab initio density functional theory Becke3P86/6-311G** treatment. Assignments of the vibrational modes are proposed that consider the results of these calculations and the observed spectra. The results of the ab initio treatment are related to crystallographic and spectral data, and they are compared with previous assignments for similar molecules. PMID- 10805917 TI - Intercellular junctions between palisade nerve endings and outer root sheath cells of rat vellus hairs. AB - Hair follicles have a longitudinal set of sensory nerve endings called palisade nerve endings (PN). We examined the junctional structures between the PN and outer root sheath (ORS) cells of hair follicles in the rat external ear. Transmission electron microscopy of serial thin sections showed that the processes of the ORS cells penetrated the basal lamina of the hair follicle, forming intercellular junctions with the PN (PN-ORS junctions). Two types of junctions were found: junctions between nerve endings and ORS cells (N-ORS junctions) and those between Schwann cell processes and ORS cells (S-ORS junctions). The N-ORS junctions had two subtypes: 1) a short process or small eminence of the ORS cell was attached to the nerve ending (type I); or 2) a process of the ORS cell was invaginated into the nerve ending (type II). The S ORS junctions also had two subtypes: 1) a short process or small eminence of the ORS cell was abutted on the Schwann cell process (type I); or 2) a process of the ORS cell was invaginated into the Schwann cell process (type II). Vesicles, coated pits, coated vesicles, and endosomes were sometimes seen in nerve endings, Schwann cells, and ORS cells near the junctions. Computer-aided reconstruction of the serial thin sections displayed the three-dimensional structure of these junctions. These results suggested that the PN-ORS junctions provided direct relationships between the PN and ORS in at least four different patterns. The discovery of these junctions shows the PN-ORS relationship to be closer than previously realized. We speculate that these junctions may have roles in attachment of the PN to the ORS, contributing to increases in the sensitivity of the PN, and in chemical signaling between the PN and ORS. PMID- 10805918 TI - Pattern of calretinin immunoreactivity in the main olfactory system and the vomeronasal system of the tree shrew, Tupaia belangeri. AB - The distribution of the calcium-binding protein calretinin was studied in peripheral and central parts of the main olfactory system (MOS) and the vomeronasal system (VNS) of adult tree shrew Tupaia belangeri. The calretinin immunoreaction was carried out with a peroxidase-coupled polyclonal antibody. In the VNS, complete labeling of all receptor cells and vomeronasal nerve fibers was observed, whereas only a subset of the somata and dendrites of receptor cells and of the olfactory nerve fibers of the MOS was immunoreactive. From the immunoreactive dendritic clubs of vomeronasal receptor cells, calretinin-labeled structures, presumably clumps of microvilli, arose that terminated within immunopositive portions of the mucus. In the main olfactory bulb, the neuropil of some of the glomeruli was immunoreactive. All periglomerular and many mitral cells were labeled. The external plexiform layer was subdivided into a faintly immunoreactive superficial half and a strongly immunoreactive deep half. Immunoreactive basal dendrites of mitral cells could be followed into either the deep half or the superficial half. In the laminated internal granular layer, a subset of immunopositive granule cells extended dendrites into the external plexiform layer. Mitral cells and granule cells with dendrites ascending to different levels of the external plexiform layer may represent functional subclasses. In the accessory olfactory bulb, all vomeronasal nerve fibers, glomeruli, and mitral/tufted cells were labeled, whereas immunoreactive periglomerular cells and internal granule cells were only scattered. In Tupaia, calretinin immunoreactivity is a more general property of the primary projecting neurons of the VNS than of the MOS and possibly indicates the involvement of calretinin in the perception of certain of the olfactory qualities. PMID- 10805919 TI - Retinal cell addition and rod production depend on early stages of ocular melanin synthesis. AB - Retinal mitosis is regulated by dopa, a melanin precursor present in the developing retinal pigment epithelium. Its absence results in retinal deficits including a failure of approximately 30% of the rod population to develop. Here, (3)H-thymidine labelling is used to analyse patterns of cell addition spanning the main period of retinal development in rat litters containing both pigmented and albino phenotypes. Many more thymidine-labelled cells are found in each cellular layer at maturity in albinos than in their pigmented littermates. Normal spatial patterns of photoreceptor addition are seen in albinos during cone production and for most of the subsequent period of rod addition. However, abnormal spatial patterns of cell addition occur across the retinal when rod production peaks. A delay in the centre to periphery gradient of cell addition is apparent in both nuclear layers. These data are related to deficits in the mature architecture of the albino retina. The results are consistent with there being significant cell cycle and/or exit point irregularities in hypopigmented retinae. It is probable that reduced dopa levels in albinos result in the cell cycle rate not slowing appropriately with development, which may lead to cells missing their exit points. This produces abnormal patterns of cell addition at key stages and delays in the gradient of retinal maturation along with a large cell loss at critical stages of rod production. PMID- 10805920 TI - Immunocytochemical basis for a meningeo-glial network. AB - Evidence is presented here for a cellular network that courses through all layers of meninges, the vasculature of both the brain and meninges, and extends into the brain parenchyma. Confocal mapping of calcium-binding protein S100beta immunoreactivity (S100beta-ir) and of the intermediate filament vimentin-ir through serial sections of the meningeal-intact adult rat brain revealed this network. In all tissues examined, S100beta-ir and vimentin-ir were primarily colocalized, and were found in cells with elongated processes through which these cells contacted one another to form a network. The location of labeling and the morphology of the cells labeled were consistent with the possibility that this network consists of fibroblasts in the meninges and the walls of large blood vessels, of pericytes at the level of capillaries, and of ependymocytes and a population of astrocytes in the brain parenchyma. At many sites along the borders of the brain parenchyma itself and of the brain blood vessels, it was possible to detect S100beta-ir and vimentin-ir cell processes that cross the basal laminae. This suggested the probable means by which the S100beta-ir cells of the extraparenchymal tissues anatomically contact the cells that express the same markers in the brain. Privileged anatomical relationships of the S100beta/vimentin network with the glial fibrillary acidic protein (GFAP) astrocytes further suggested that, together, they form the structural basis for a general meningeo-glial network. This organization challenges the current model of brain architecture, calls for a reconsideration of the role of meninges and vascular tissues, and appears to reflect the existence of hitherto unsuspected systems of communication. PMID- 10805921 TI - Identification of multiple somatostatin receptors in the rat somatosensory cortex during development. AB - The present study was aimed at identifying somatostatin receptor subtypes on the basis of their ligand-binding properties in the rat somatosensory cortex during fetal and postnatal development. Characterization of somatostatin-binding sites was performed in individual cortical layers by using three radioligands and eight competitors with known selectivities for the five somatostatin receptor subtypes. Binding sites sensitive to sst2-selective ligands were detected with high densities in the intermediate zone of the fetal cortex. From embryonic day 21 to 21 days postnatal (P21), mixed populations of receptors were detected in the cortical plate and emerging layers I-VI. Putative sst2 receptors were detected throughout the entire period but displayed different affinities for somatostatin and analogs, and a different sensitivity to GTP, depending on the developmental stage and the cortical layer considered. High densities of binding sites exhibiting characteristics of the sst1, sst3/5, and sst4 receptor subtypes were observed from P4 to P7, P7 to P14, and P7 to P21, respectively. In addition, each type of site exhibited a particular distribution pattern across the cortical layers that varied during the development. In the adult cortex, binding sites with sst1 and sst2 receptor characteristics were predominant. This study provides evidences of developmental expression windows of four sst receptor subtypes in selected areas of the rat cerebral cortex. PMID- 10805922 TI - Colocalization of multiple GABA(A) receptor subtypes with gephyrin at postsynaptic sites. AB - Clustering of gamma aminobutyric acid (GABA)(A) receptors to postsynaptic sites requires the presence of both the gamma2 subunit and gephyrin. Here, we analyzed by double-immunofluorescence staining the colocalization of gephyrin and major GABA(A)-receptor subtypes distinguished by the subunits alpha1, alpha2, alpha3, or gamma2 in adult rat brain. By using confocal laser scanning microscopy, GABA(A)-receptor subunit staining revealed brightly stained clusters that were colocalized with gephyrin-positive clusters of similar size and distribution in several brain regions, including cerebellum, hippocampus, thalamus, and olfactory bulb. In addition, a diffuse staining was observed for GABA(A)-receptor subunits in the neuropil, presumably representing extrasynaptic receptors. Overall, only few gephyrin-positive clusters were not colocalized with GABA(A)-receptor subunit clusters. Electron microscopic analysis in cerebellar cortex confirmed the selective postsynaptic localization of gephyrin. High-resolution images (voxel size, 50 x 50 x 150 nm) were restored with an iterative image deconvolution procedure based on a measured point-spread function to analyze the colocalization between GABA(A)-receptor subunits and gephyrin in individual clusters. This analysis revealed a considerable heterogeneity in the micro-organization of these presumptive GABAergic postsynaptic sites. For instance, whereas gephyrin- and gamma2 subunit-positive clusters largely overlapped in the cerebellar molecular layer, the colocalization was only partial in glomeruli of the granule cell layer, where small gephyrin clusters typically were "embedded" in larger GABA(A) receptor clusters. These findings show that gephyrin is associated with a majority of GABA(A)-receptor subtypes in brain, and document the usefulness of image deconvolution for analyzing the structural organization of the postsynaptic apparatus by fluorescence microscopy. PMID- 10805923 TI - Roles of glutamate and FMRFamide-related peptides at the chromatophore neuromuscular junction in the cuttlefish, Sepia officinalis. AB - Body patterning behavior, the expression of highly intricate patterns, is ubiquitous among all unshelled cephalopods. These body patterns are in part generated by the coordinated activity of millions of skin chromatophore organs, each of which is regulated by a set of chromatophore muscles directly innervated by centrally located chromatophore motoneurons. This study addresses the question of the identity and function of the transmitter(s) at the chromatophore neuromuscular junction (NMJ) in the European cuttlefish Sepia officinalis. Glutamate application causes a rapid contraction of the chromatophore muscles, resulting in chromatophore expansion. Pharmacological studies demonstrate that the chromatophore muscles contain receptors blocked by glutamate-specific antagonists. Glutamate-like immunoreactivity is also present in the somata of putative chromatophore motoneurons. These findings suggest that glutamate likely acts as a neurotransmitter at the chromatophore NMJ. Evidence is also presented suggesting that FMRFamide-related peptides (FaRPs) also function as neurotransmitters at the Sepia chromatophore NMJ. FMRFamide application causes contraction of chromatophore muscles; however, the FMRFamide effect is slower and longer lasting than that of glutamate. Pharmacological data show that FMRFamide acts directly on the chromatophore muscles. FMRFamide-immunopositive cells are present in the posterior chromatophore lobe, the putative location of the chromatophore motoneuron somata. A combination of immunocytochemistry and in situ hybridization shows that some putative chromatophore motoneurons express FaRP like immunoreactivity and an FaRP-coding mRNA transcript. Many FMRFamide immunopositive cells in the posterior chromatophore lobes also express glutamate like immunoreactivity. We conclude that glutamate and FaRPs likely function as fast and slow transmitters, respectively, at the Sepia chromatophore NMJ. PMID- 10805924 TI - Localization of choline acetyltransferase in the developing and adult turtle retinas. AB - Acetylcholine has important epigenetic roles in the developing retina. In this study, cells that expressed choline acetyltransferase (ChAT), the enzyme that synthesizes acetylcholine, were investigated in embryonic, postnatal, and adult turtle retinas by using immunofluorescence histochemistry. ChAT was present at stage 15 (S15) in cells near the vitreal surface. With the formation of the inner plexiform layer (IPL) at S18, ChAT-immunoreactive (-IR) cells were located in the inner nuclear layer (INL) and the ganglion cell layer (GCL). In the INL, presumed starburst amacrine cells were homogenous in appearance and formed a single row next to the IPL: This pattern was conserved until adulthood. In the GCL, however, there were multiple rows of ChAT-IR cells early in development, and this high density of labeled cells continued during the embryonic stages, until around birth. The high density of ChAT-IR cells in the GCL was due in part to a population of cells that expressed ChAT transiently. In postnatal stages and adult retinas, the presumed starburst amacrine ChAT-IR cells formed two mirror like rows of homogenous cells on both borders of the IPL. Two cholinergic dendritic strata that were continuous with these cells were observed as early as S18, and their depths in the IPL were relatively stable throughout development. A third population of ChAT-IR cells was observed toward the middle of the INL around S25 and persisted into adulthood. Finally, cells in the outer nuclear layer (ONL) were ChAT-IR during the embryonic stages, were less immunoreactive during the postnatal stages, and were not immunoreactive in the adult retinas. PMID- 10805925 TI - Colocalization of choline acetyltransferase and gamma-aminobutyric acid in the developing and adult turtle retinas. AB - Acetylcholine and gamma -aminobutyric acid (GABA) are putative neurotransmitters in the adult vertebrate retina. In this study, cells that coexpress choline acetyltransferase (ChAT) and GABA or glutamic acid decarboxylase (GAD) were investigated in turtle retinas from stage 14 (S14) to adulthood by using a double labeling immunofluorescence technique. ChAT immunoreactivity was observed at S15 and included not only the presumptive starburst cholinergic amacrine cells but also a population in the ganglion cell layer (GCL) that expressed ChAT transiently during the embryonic stages (see the accompanying paper: Nguyen et al. [2000] J. Comp. Neurol. 420:512-526). PMID- 10805926 TI - Impairment in motion discrimination tasks is unrelated to amount of damage to superior temporal sulcus motion areas. AB - The behavioral role of the middle temporal (MT/V5) area and its satellites in motion processing is still unclear, particularly the degree to which MT/V5 proper is critical for different types of motion processing. Therefore, effects of small and large lesions in the caudal part of the superior temporal sulcus of macaque monkeys were compared for two tasks requiring different types of motion processing: a direction and a kinetic orientation discrimination. The small lesion was restricted to the peripheral representation of MT/V5 but included V4t, whereas the large lesion included all of MT/V5 and the medial superior temporal (MST) area as well as substantial parts of the floor of the superior temporal (FST) area. Both lesions resulted in significant and long-lasting impairment of direction discrimination but had a lesser effect on kinetic orientation discrimination. Thus the effects of small STS lesions on motion perception are much stronger than expected. PMID- 10805927 TI - Everything you never wanted to know about polarization--but were afraid you might find out. PMID- 10805928 TI - Polarization of scatter and fluorescence signals in flow cytometry. AB - BACKGROUND: The pulses of light scatter and fluorescence measured in flow cytometers exhibit varying degrees of polarization. Flow cytometers are heterogeneously sensitive to this polarization, depending on the light source(s), the optical layout, and the types of mirrors and filters used. Therefore, fluorescence polarization can affect apparent intensity ratios between particles and interfere with schemes for interlaboratory standardization. METHODS: We investigate the degree to which polarization affects common flow cytometry measurements. Our technique for determining polarization differs from previous methods because complete distributions of intensity versus polarization angle are measured, rather than intensities at just two orthogonal polarization angles. Theoretical models for scatter and fluorescence are presented and verified by making polarization measurements of calibration beads. RESULTS: Measurements of cells stained with a variety of dyes illustrate that fluorescence polarization occurs frequently in flow cytometry. CONCLUSIONS: Consequences for quantitative cytometry are discussed, and the use of the "magic angle" to make a flow cytometer insensitive to fluorescence polarization is proposed. PMID- 10805929 TI - Suspension arrays for high throughput, multiplexed single nucleotide polymorphism genotyping. AB - BACKGROUND: Genetic diversity can help explain disease susceptibility and differential drug response. The most common type of variant is the single nucleotide polymorphism (SNP). We present a low-cost, high throughput assay for SNP genotyping. METHODS: The assay uses oligonucleotide probes covalently attached to fluorescently encoded microspheres. These probes are hybridized directly to fluorescently labeled polymerase chain reaction (PCR) products and the results are analyzed in a standard flow cytometer. RESULTS: The genotypes determined with our assay are in good agreement with those determined by TaqMan. The range of G/C content for oligonucleotide probes was 23.5-65% in the 17 bases surrounding the SNP. Further optimization of probe length and target concentration is shown to dramatically enhance the assay performance for certain SNPs. Using microspheres which have unique fluorescent signatures, we performed a 32-plex assay where we simultaneously determined the genotypes of eight different polymorphic genes. CONCLUSIONS: We demonstrate, for the first time, the feasibility of multiplexed genotyping with suspension arrays using direct hybridization analyses. Our approach enables probes to be removed from or added to an array, enhancing flexibility over conventional chips. The ability to multiplex both the PCR preparation and the hybridization should enhance the throughput, cost, and speed of the assay. PMID- 10805930 TI - Extension of osmolality-induced podia is observed from fluorescently labeled hematopoietic cell lines in hyperosmotic medium. AB - BACKGROUND: Since the description of long podia extended by hematopoietic cells and cell lines, the reliable elicitation of podia extensions is needed to study these podia systemically. In this study, hyperosmotic stress was considered as an elicitor. METHODS: Using two fluorescent membrane dyes PKH2 and PKH26, and an automated fluorescence microscopy system, morphological changes of seven human cell lines (six hematopoietic, one fibrosarcoma) at different osmolalities were monitored. Presence of surface molecules on the hyperosmolality-induced podia (osmopodia) was examined. RESULTS: In hyperosmotic medium, cells shrank rapidly, followed by osmopodia extension. Cells exhibited variable number (up to five) and length (up to longer than 100 microm) of osmopodia in about 1 h. Dead cells did not extend podia. Frequency, length, and number of podia were variable among cell lines studied. CD44 and CD45 were not present on the osmopodia, although they were present on the cell surface, showing that osmopodia characteristics differ from the podia observed previously in isotonic media. The osmopodia extension process was shown to be reversible upon repeated osmolality changes. CONCLUSIONS: Osmopodia extended by human hematopoietic cell lines display a newly observed cellular morphology and provide a tool for investigation of dynamic cellular response to environmental changes. PMID- 10805931 TI - Intercellular adhesion can be visualized using fluorescently labeled fibrosarcoma HT1080 cells cocultured with hematopoietic cell lines or CD34(+) enriched human mobilized peripheral blood cells. AB - BACKGROUND: Intercellular contacts between adjacent cells migrating over each other are important in many cellular processes. However, it has been difficult to visualize and identify dynamic intercellular adhesions between migrating cells in situ. METHODS: Two fluorescent membrane dyes, PKH2 and PKH26 for staining HT1080 and hematopoietic cells and cell lines, and an automated fluorescence microscopy system were used to monitor intercellular adhesion. RESULTS: Cellular extensions connecting two or more adjacent cells were visualized, showing the intercellular adhesion between migrating cells for minutes and up to hours. After cells adhered to each other, followed by cell migration in different directions, cellular extensions were dragged from the pivotal contact points in different focal planes. CD34(+)-enriched mobilized peripheral blood cells and six hematopoietic cell lines showed intercellular connections in cocultures with HT1080. However, the frequency of intercellular connections was variable in different cocultures. A cell density of about 3.1 x 10(4) cells/cm(2) for both cell lines in cocultures provided an adequate number of cells in each field of view, showing up to four intercellular connections per 100 total cells plated. DISCUSSION: The tools derived from this study will open new areas of investigation for understanding the mechanism of the intercellular adhesion process. PMID- 10805932 TI - Simultaneous flow cytometric analyses of enhanced green and yellow fluorescent proteins and cell surface antigens in doubly transduced immature hematopoietic cell populations. AB - BACKGROUND: Cell transduction with multiple genes offers opportunities to investigate specific gene interactions on cell function. Detection of multiple transduced genes in hematopoietic cells requires strategies to combine measurements of gene expression with phenotypic cell discriminants. We describe simultaneous flow cytometric detection of two green fluorescent protein (GFP) variants in immunophenotypically defined human hematopoietic subpopulations using only a minor physical adjustment to a standard FACSCalibur. METHODS: The accuracy and sensitivity of enhanced GFP (EGFP) and enhanced yellow fluorescent protein (EYFP) detection in mixtures of transduced and nontransduced PG13 packaging cells were evaluated by flow cytometry. Retroviral vectors encoding EGFP or EYFP were used to transduce CD34(+) hematopoietic cells derived from umbilical cord blood. The transduction efficiency into subpopulations of hematopoietic cells was measured using multivariate flow cytometry. RESULTS: A bicistronic retroviral vector containing the EGFP and puromycin N-acetyltransferase (pac) genes afforded brighter EGFP signals in transduced cells than a retroviral vector encoding a pac EGFP fusion protein. The sensitivity of detecting EGFP and EYFP-expressing cells among a background of nonexpressing cells was 0.01% and 0.05%, respectively. EGFP or EYFP was expressed in up to 95% of CD34(+) DR(-) or CD34(+) 38(-) subpopulations in cord blood 48 h posttransduction. Simultaneous transduction with EGFP and EYFP viral supernatants (1:1 mixture) led to coexpression of both GFP variants in 15% of CD34(+) DR(-) and 20% of CD34(+) 38(-) cells. CONCLUSIONS: These results demonstrate simultaneous detection of EGFP and EYFP in immunophenotypically discriminated human hematopoietic cells. This technique will be useful to quantify transduction of multiple retroviral constructs in discriminated subpopulations. PMID- 10805933 TI - Susceptibility of human herpesvirus 6 to antiviral compounds by flow cytometry analysis. AB - BACKGROUND: The emergence of human herpesvirus 6 (HHV-6) as a human pathogen led to the possibility of specific therapy against HHV-6 and the development of standardized susceptibility assays of HHV-6 to antivirals. METHODS: We have developed a flow cytometry method to analyze the multiplication of the HST strain of human herpesvirus 6 (HHV-6) variant B in vitro using monoclonal antibodies specific to virus proteins. This method was subsequently used to determine the sensitivity of HST multiplication in MT4 cells to four antiviral compounds of three different classes: acyclovir (ACV) and ganciclovir (GCV), two acyclic guanosine analogs; cedofovir (CDV), an acyclic nucleoside phosphonate; and phosphonoformic acid (PFA), a pyrophosphate analog. RESULTS: The 50% inhibitory concentrations (IC(50)) of ACV, GCV, CDV, and PFA determined by flow cytometry assay were 25.3, 6.4, 0.95, and 6.0 microM, respectively (5.7, 1.6, 0.3, and 1.8 microg/ml, respectively). These data together with the results of cytotoxicity assays confirmed the high efficiency and selectivity of CDV and PFA against HHV-6 B in vitro, suggested by previous results. CONCLUSIONS: Our flow cytometric assay appeared as a reproducible specific method to characterize HHV-6 susceptibility to antiviral compounds. It can be considered as a convenient alternative to the other immunologic and DNA hybridization assays used for that purpose. PMID- 10805934 TI - A flow cytometric method for measuring neutralization of HIV-1 subtype B and E primary isolates. AB - BACKGROUND: Clinical trials testing candidate human immunodeficiency virus type 1 (HIV-1) vaccines have required the use of HIV neutralization assays to detect responses to specific geographic subtypes of HIV-1. The variability in results seen with current p24 neutralization assay endpoints prompted us to assess the utility of flow cytometry for monitoring the neutralization of HIV-1 primary isolates. METHODS: A modified neutralization assay was performed using CD8 depleted peripheral blood mononuclear cells (PBMC). The cells were fixed, permeabilized, stained with a directly conjugated HIV-1 p24 monoclonal antibody, and analyzed by flow cytometry. HIV-1 subtype B' and E primary isolates were tested using pooled sera or plasma from subtype B' or E infected patients. RESULTS: Primary isolate cultures (without neutralizing antibody) showed from 18% to 42% p24(+) cells, depending on the virus. Less than 0.2% p24(+) cells were detected in uninfected cultures. Subtype-specific neutralization of viruses was observed using plasma or serum pools; neutralization ranged from 0% to 99% reduction of infected cells. CONCLUSIONS: Flow cytometric detection of intracellular HIV-1 p24 can be used as an endpoint assay to assess neutralization of HIV-1 subtypes B' and E primary isolates. This enumerative method has the advantage of identifying intracellular p24 in specific subsets at an early culture timepoint. It also provides an alternative quantitative endpoint for HIV neutralization assays. PMID- 10805935 TI - Flow cytometry detection of caspase 3 activation in preapoptotic leukemic cells. AB - BACKGROUND: Procaspase 3 is a constitutive proenzyme that is activated by cleavage during apoptosis. The resulting enzyme is able to cleave several target proteins after the second aspartate of a DEVD sequence common to all the substrates of caspases 3 and 7 (DEVDase). Because active caspase 3 is a common effector in several apoptotic pathways, it may be a good marker to detect (pre )apoptotic cells by flow cytometry (FCM). Materials and Methods Apoptosis was induced in U937 or bone marrow mononuclear cells by daunorubicin (DNR), idarubicin (IDA), or camptothecin (CAM). Viable and apoptotic cells were sorted by FCM on the basis of either fluorescein isothiocyante (FITC)-annexin V binding or DiOC6(3) accumulation. DEVDase activity was measured in sorted populations by spectrofluorometry. Cleaved caspase 3 was labeled in situ with phycoerythrin (PE) conjugated anti-activated caspase 3 antibodies and analyzed by FCM. RESULTS: DEVDase activity was detected in sorted viable CAM- and DNR-treated U937 cells, demonstrating that a partial caspase activation occurred earlier than phosphatidyl-serine exposure and mitochondrial membrane potential dissipation. The presence of a low amount of active caspase 3 in the treated viable cells was confirmed in situ with PE-conjugated anti-active caspase 3 antibodies. The same antibody was used in combination with FITC-annexin V and CD45-PC5 to study caspase 3 activation in acute leukemia blast cells after in vitro DNR and IDA treatment. Both anthracyclines induced a caspase 3-dependent apoptosis that was more efficient in blast cells than in contaminating lymphocytes. CONCLUSIONS: These results demonstrate that anti-active caspase 3 labeling can be an alternative to fluorogenic substrates to efficiently detect early apoptosis by FCM in heterogeneous samples. They also confirm that anthracyclines induce blast cell apoptosis by a caspase 3-dependent pathway. PMID- 10805936 TI - Relationship between cytogenetic aberrations by CGH coupled with tissue microdissection and DNA ploidy by laser scanning cytometry in head and neck squamous cell carcinoma. AB - BACKGROUND: The relationship between DNA sequence copy number aberrations (DSCNAs) and DNA ploidy in head and neck squamous cell carcinomas (HNSCCs) is still controversial. Materials and Methods We analyzed DSCNAs by comparative genomic hybridization (CGH) combined with microdissection and DNA ploidy by laser scanning cytometry (LSC) in 18 surgically removed HNSCCs and compared the data. RESULTS: Copy number increases were most frequently observed on chromosomes 3q (16 cases), 8q (13 cases), and 12p (11 cases). Copy number decreases were observed on chromosome 3p (14 cases). LSC revealed DNA aneuploidy in 10 of the 18 cases. All DNA aneuploid tumors exhibited gain or amplification of DNA copy number at 12p11-12.1, whereas gain of DNA copy number was found in only 1 of 8 diploid tumors. DSCNAs were more frequent in DNA aneuploid tumors than in diploid tumors (P < 0.005). CONCLUSIONS: The present observations indicate a close relationship between DSCNAs and DNA ploidy in HNSCCs. PMID- 10805937 TI - A new four-color flow cytometric assay to detect apoptosis in lymphocyte subsets of cultured peripheral blood cells. AB - BACKGROUND: Human peripheral blood lymphocytes kept in culture after isolation die by an apoptotic process. Detection of apoptosis with labeled Annexin V to demonstrate loss of plasma membrane asymmetry is sensitive, specific, and easy using flow cytometry. This is true in lymphoblastic cell lines when combining Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI). However, measurement of apoptosis by flow cytometry in isolated human lymphocytes using Annexin V-FITC/PI is disturbed by the presence of a variable percentage of erythrocytes in the isolated lymphocyte population. To overcome this problem, we have developed and tested a new four-color flow cytometric assay to detect apoptosis in lymphocyte subsets of cultured peripheral blood cells. METHODS: Peripheral blood lymphocytes are isolated by density gradient centrifugation. Nucleus-containing cells are selected using CD45-phycoerythrin (PE). The lymphocyte subset of interest is selected using CD4, CD8, or CD19 energy-coupled dye (ECD) labeling. Apoptosis is detected using Annexin V-FITC with 7-amino Actinomycin-D (7-AAD) to distinguish early apoptotic from late apoptotic lymphocytes. RESULTS: We have developed a new technique to detect apoptosis in isolated human peripheral blood lymphocyte subsets with good reproducibility, coefficient of variation < 17%. CONCLUSIONS: We now have a validated tool to study apoptosis in subsets of isolated human lymphocytes to increase our knowledge of pathogenesis and therapies in lymphoreticular malignancies. PMID- 10805938 TI - The efficacy of long-term epilation of unwanted hair by noncoherent filtered flashlamp. PMID- 10805939 TI - Optimal parameters for laser tissue soldering: II. Premixed versus separate dye solder techniques. AB - BACKGROUND AND OBJECTIVE: Laser tissue soldering by using an indocyanine green (ICG)-doped protein solder applied topically to the tissue surface and denatured with a diode laser was investigated in Part I of this study. The depth of light absorption was predominantly determined by the concentration of the ICG dye added to the solder. This study builds on that work with an in vitro investigation of the effects of limiting the zone of heat generation to the solder-tissue interface to determine whether more stable solder-tissue fusion can be achieved. STUDY DESIGN/MATERIALS AND METHODS: An alternative laser tissue soldering technique was investigated, which increased light absorption at the vital solder tissue interface. A thin layer of ICG dye was smeared over the surface to be treated, the protein solder was then placed directly on top of the dye, and the solder was denatured with an 808-nm diode laser. Because laser light at approximately 800 nm is absorbed primarily by the ICG dye, this thin layer of ICG solution restricted the heat source to the space between the solder and the tissue surfaces. A tensile strength analysis was conducted to compare the separate dye-solder technique with conventional techniques of laser tissue soldering for which a premixed dye-solder is applied directly to the tissue surface. The effect of hydration on bond stability of repairs formed by using both techniques was also investigated using tensile strength and scanning electron microscopy analysis. RESULTS: Equivalent results in terms of tensile strength were obtained for the premixed dye-solder technique using protein solders containing 0.25 mg/ml ICG (liquid solder, 220 +/- 35 N/cm(2); solid solder, 602 +/- 32 N/cm(2)) and for the separate dye-solder technique (liquid solder, 228 +/- 41 N/cm(2); solid solder, 578 +/- 29 N/cm(2)). The tensile strength of native bovine thoracic aorta was 596 +/- 31 N/cm(2). Repairs created by using the separate dye-solder technique were more stable during hydration than their premixed dye-solder counterparts. The conventional premixed dye-solder was simpler and approximately twice as fast to apply. The separate dye-solder technique, however, increased the shelf-life of the solder, because the dye was mixed at the time of the experiment, thus conserving its spectral absorbency properties. CONCLUSION: Two laser-assisted tissue soldering techniques have been evaluated for repairing aorta incisions in vitro. The advantages and disadvantages of each of these techniques are discussed. PMID- 10805940 TI - Laser modulation of angiogenic factor production by T-lymphocytes. AB - BACKGROUND AND OBJECTIVE: In previous investigations, small variations in the energy densities of low level light therapy (LLLT) were found to produce significant differences in the proliferation of resting T-lymphocytes in vitro. Pulsing these cells with mitogen in addition to laser therapy produced inhibitory effects regardless of the amplitude of the energy density used. In the current study, the effect of LLLT on the production of angiogenic factor(s) by T lymphocytes was investigated in vitro. STUDY DESIGN/MATERIALS AND METHODS: Human T-cells isolated from peripheral blood were prepared in suspension either with or without addition of mitogen. Cell suspensions were irradiated with laser by using the following energy densities: 1.2, 3.6, 6.0, and 8.4 J/cm(2). Wavelength, pulsing frequency, and power output were kept constant at 820 nm, 5,000 Hz, and 50 mW, respectively. After either 3 or 5 days of incubation, lymphocyte supernatants were collected and added as conditioned media to cultured endothelial cells (ECs). The effect on the proliferation of these ECs was assessed over a 72-hour period by using a methylene blue assay. RESULTS: Endothelial cell proliferation increased significantly when incubated with conditioned media collected from resting T-cells exposed to 1.2 and 3.6 J/cm(2). Day 5 conditioned media produced similar patterns of EC proliferation to that of day 3 but at lower magnitude. Pulsing of T-lymphocytes with mitogen in addition to laser irradiation significantly lessened their angiogenic capability. Conditioned media from 3.6 J/cm(2) laser-treated T-cells induced the maximal EC proliferation in all groups studied. CONCLUSION: It would seem that laser therapy stimulates lymphocytes to produce factor(s) that can modulate EC proliferation in vitro; this effect on the lymphocytes is influenced by (1) the amplitude of energy density used for T-cell irradiation, (2) exposing T-cells to both mitogen and laser, and (3) the duration of T-cell incubation in culture. PMID- 10805941 TI - Laser-induced shock wave endothelial cell injury. AB - BACKGROUND AND OBJECTIVE: Several laser procedures, extracorporeal lithotripsies (ESWL), and high-velocity missile trauma generate pressure transients that are transmitted through the tissues. Despite several publications demonstrating shock wave-induced tissue injury, little is known about its pathophysiology. This study introduces an in vitro model for studying shock wave effects on endothelial cell (EC) monolayers. STUDY DESIGN/MATERIALS AND METHODS: A Nd:YAG laser-driven flyer plate technique was used to generate shock waves. Physical characteristics were determined with a pressure transducer, a high-speed video camera, and sequential photography. Biological effects were studied with phase contrast and lightfield microscopy, computerized morphometry, immunocytochemistry, spectrophotometry, and enzyme-linked immunosorbent assay (ELISA). RESULTS: The shock waves generated were highly reproducible. Cavitation was verified and quantified, and its extent could be varied in the vials. Exposed cultures exhibited areas with cell membrane damage and cell detachment. Release of LD was elevated (P < 0.01) in exposed vials. The EC lesions were larger (>P < 0.01) in cultures submitted to high vs. low extent of cavitation. CONCLUSION: The flyer-plate model can be used to subject cell monolayers to defined and reproducible shock waves causing immediate cell injury similar to the previously reported vascular lesions associated with ESWL, pulsed lasers, and blast trauma. With the flyer-plate model, such lesions may be further studied on the cellular and subcellular levels. PMID- 10805942 TI - Q-switched Nd:YAG laser treatment for acquired bilateral nevus of ota-like maculae: a long-term follow-up. AB - BACKGROUND AND OBJECTIVE: Acquired bilateral nevus of Ota-like maculae (ABNLM) is not an uncommon dermal melanosis in Asian people. It is known for its recalcitrance to conventional treatment. We proposed Q-switched Nd:YAG laser for the treatment of this entity. STUDY DESIGN/PATIENTS AND METHODS: Seventy female patients with ABNLM, who were 25-40 years old (mean, 37 years), were included in a prospective clinical study. Q-switched Nd:YAG laser at fluence of 8-10 J/cm(2), spot size 2 or 4 mm, and a repetition rate of 10 Hz was used to treat the lesions. RESULTS: Two patients were lost to follow-up. In the remaining 68 cases, all lesions attained a 100% clearance after two to five treatment sessions (mean, 2.8 sessions). The results had persisted at 3-4 years follow-up (mean, 42 months). Temporary hyperpigmentation was found in 50% of patients; there was no scarring or changing of skin texture. CONCLUSION: Q-switched Nd:YAG laser is a safe and effective noninvasive alternative treatment for ABNLM. PMID- 10805943 TI - Nd:YAG laser surgery for the excision of pilonidal cysts: a comparison with traditional techniques. AB - BACKGROUND AND OBJECTIVE: Nd:YAG laser photothermal ablation has been accepted as a treatment modality for hemorrhoidal disease. There is little reported on its use in treating pilonidal disease. We hypothesized that laser would be an excellent tool for pilonidal cystectomy, facilitating improved outcome and patient satisfaction. STUDY DESIGN/MATERIALS AND METHODS: A 5-year retrospective study was performed comparing Nd:YAG laser to the standard surgical technique. A telephone questionnaire addressing the length of time the cyst was debilitating both preoperatively and postoperatively as well as length of convalescent time before return to work was administered. Pain was assessed by using an analog pain scale. RESULTS: Operative time for the traditional pilonidal cystectomy was 20 minutes longer than Nd:YAG laser cystectomy. Postoperative hospital stay was similar. Laser patients returned to work an average of 2.4 days earlier, and their postoperative pain was less than those treated traditionally. CONCLUSION: In an era when the medical consumer makes decisions based on the efficacy of treatment by using criteria such as pain, length of hospitalization, and speed of return to work, Nd:YAG lasers have emerged as a surgical tool that can fulfill these criteria for certain procedures. Patient postoperative satisfaction after laser excision was greater when compared with those who had traditional excisions. Postoperative pain was less, as was the pain experienced during the first week of recovery. Cost for both was comparable. PMID- 10805944 TI - In vitro temperature change at the dentin/pulpal interface by using conventional visible light versus argon laser. AB - BACKGROUND AND OBJECTIVE: The argon laser has been promoted as a competing technology to multi-wavelength visible light as a curing source for dental restorative resins. However, the comparative thermal risk to the pulp between these two sources of light energy requires determination. The objective of this study is to compare the temperature induced at the dentin-pulpal interface between the argon laser and visible light curing unit at a variety of exposure regimens and conditions. STUDY DESIGN/MATERIALS AND METHODS: In vitro temperatures were measured and recorded at the dentin-pulpal interface upon external light exposure. Independent variables included the dentin thickness, duration and waveform of exposure, and presence of composite resin. RESULTS: In most instances, the argon laser resulted in less temperature rise on the pulpal dentin interface. CONCLUSION: The argon laser should not pose a serious thermal risk to the pulp if used at recommended energies. PMID- 10805945 TI - Pulsed Nd:YAG laser irradiation of the tooth pulp in the cat: I. Effect of spot lasing. AB - BACKGROUND AND OBJECTIVE: The purpose of the present study was to evaluate physiologically pulpal nerve responses and to elucidate histopathologically the pulp tissue reactions to "spot irradiation" with a pulsed Nd:YAG laser. STUDY DESIGN/MATERIALS AND METHODS: Antidromic compound action potentials and the pulpal blood flow (PBF) were recorded from the canine tooth of a sodium pentobarbitone-anesthetized cat. The laser irradiation-induced pulp tissue changes were histologically investigated. RESULTS: The coronal antidromic compound action potentials disappeared in all the teeth tested during lasing, and the time needed to erase them was significantly shortened with increases in lasing power (P < 0.05). The radicular PBF increased when spot irradiation was performed, and the coronal PBF also temporarily increased with low-powered lasing. Histologic investigation revealed that spot irradiation with the laser produced severe damage in the pulp tissue in a dose-dependent manner. CONCLUSION: The present study suggests that spot irradiation with a pulsed Nd:YAG laser risks producing nerve injury and irreversible tissue damage in the pulp with lasing for the purpose of desensitizing hypersensitive dentin. PMID- 10805946 TI - A study of aminolevulinic acid-induced protoporphyrin IX fluorescence kinetics in the canine oral cavity. AB - BACKGROUND AND OBJECTIVE: 5-Aminolevulinic acid-induced protoporphyrin IX is a promising photosensitizer that could enhance the spectroscopic contrast between normal and diseased oral tissues. Knowledge of the pharmacokinetics and effects on tissue type are important for diagnostic and therapeutic procedures. STUDY DESIGN/MATERIALS AND METHODS: Dogs randomly were administered five doses of 5 aminolevulinic acid: 5, 25, 50, 75, and 100 mg/kg. The fluorescence was recorded from buccal mucosa, gums, tongue, and facial skin using a fiberoptic probe connected to an optical multichannel analyzer. Blood samples were collected for hematologic and serum biochemical analysis. Pharmacokinetic parameters of interest were estimated using a compartmental model. RESULTS: Protoporphyrin fluorescence at all sites reached a peak in 2-6 hours, and returned to baseline in 24-31 hours, depending on the dose. Plasma protoporphyrin peaked earlier than oral tissues. CONCLUSION: The rate of synthesis of protoporphyrin, and its conversion to heme products are dose dependent. Different tissues have different pharmacokinetic response. PMID- 10805947 TI - Corrigendum PMID- 10805948 TI - Cancer genetics and the surgeon PMID- 10805949 TI - Genes implicated in hereditary breast cancer syndromes. AB - The medical histories of breast cancer-prone families have been described for over a century. The pattern of breast cancer occurrences in these families is most consistent with an autosomal dominant mode of inheritance. The location of a gene that could explain the pattern of transmission of the breast cancer trait in families averaging early (pre-menopausal) onset of breast cancer was reported in 1990. Since then, two genes have been identified: BRCA1 and BRCA2. Germ-line mutations in these two genes confer susceptibility to breast (female and male) and ovarian cancer, and account for a significant proportion of hereditary breast cancer in two cancer syndromes: site-specific breast cancer and the breast ovarian cancer syndrome. Other hereditary syndromes that feature breast cancer are Li-Fraumeni syndrome, Cowden disease, and ataxia telangiectasia, whose carriers have been shown to harbor germ-line mutations in TP53, PTEN, and ATM, respectively. There may be other genetic factors that contribute to hereditary breast cancer, since not all families with multiple cases of breast cancer harbor germ-line BRCA1 or BRCA2 mutations. Host factors (such as lifestyle choices) and other genes may modulate risk of breast cancer in mutation carriers. PMID- 10805950 TI - Clinico-pathological characteristics of BRCA1- and BRCA2-related breast cancer. AB - Approximately 2% to 5% of all breast cancers are hereditary, meaning that the cancer predisposition is carried as a monogenic trait. Several highly penetrant breast cancer predisposing genes have been identified. These discoveries will permit a refined description of breast cancer occurring as part of the different genetic syndromes. We reviewed the medical literature on the clinico-pathological features of breast cancer associated with the major breast cancer susceptibility genes BRCA1 and BRCA2. BRCA1-associated breast cancers are more frequently ductal invasive, high-grade carcinomas with an important lymphocytic infiltration. They are aneuploid, estrogen and progesterone receptors negative, and p53 positive. BRCA2-related breast cancers tend to be higher-grade tumors than are non hereditary cases, although this association is less strong then for BRCA1 cases. These tumors exhibit substantially less tubule formation, but mitotic count and cellular pleomorphism do not differ significantly from those of sporadic cases. The overall pattern of the identified pathological characteristics suggests a carcinogenic pathway in BRCA1- and BRCA2-related breast cancers different from that found in sporadic cases. The probability of finding a BRCA1/2 germ-line mutation is partly determined by these characteristics. In addition, these features will likely influence the behavior of BRCA1/2-related breast cancer. PMID- 10805951 TI - Clinical management of BRCA1- and BRCA2-associated breast cancer. AB - The recent discovery of the breast cancer-associated genes BRCA1 and BRCA2 has changed the clinical care provided to women at high risk of breast cancer. We will review what is currently known about the clinical management of patients who bear (or are suspected of bearing) mutations in either of these two genes. The issues related to establishing a diagnosis of inherited breast cancer, deciding which women are candidates for testing, the limitations of testing, and the predictive power of these tests are addressed. The prognostic features of cancers associated with a BRCA1 and BRCA2 mutation are reviewed. Further, guidelines for prophylaxis of women with a BRCA1 or BRCA2 mutation are given and recommendations are made for the care of mutation carriers with cancer. PMID- 10805952 TI - Hereditary nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC), or Lynch syndrome, is the most common form of hereditary colorectal cancer (CRC). A well-orchestrated cancer family history is essential for its diagnosis since, unlike its familial adenomatous polyposis (FAP) hereditary cancer counterpart, HNPCC lacks distinguishing clinical stigmata of its cancer genetic risk. Discoveries in the 1990s of germ-line mutations, the most common of which are hMSH2 and hMLH1, have added enormous power to the diagnosis of Lynch syndrome. Its medical management is contingent upon its natural history. For example, approximately 70% of CRCs occur proximal to the splenic flexure, with one-third of the cancers occurring in the cecum, thereby mandating full colonoscopy. A high rate of metachronous CRCs indicates the need for no less than a subtotal colectomy for the management of initial CRC. Genetic counseling is essential prior to DNA testing, and at the time of disclosure of the results. Education of patients as well as physicians about all facets of this disorder is extremely important. If patients are to show compliance with germ-line testing, screening, and management options, they must understand the natural history and the significance of their genetic risk status. Physicians must also be aware of clinical nuances of this disorder to provide the necessary care. PMID- 10805953 TI - Familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is a dominantly inherited familial cancer syndrome characterized by an increased predisposition to colorectal cancer and other benign and malignant extra-colonic lesions. FAP has been linked to germline mutations of the adenomatous polyposis coli (APC) gene that encodes a protein with 2,843 amino acids that has important functions in the regulation of cell growth. A genotype-phenotype correlation has also been observed between mutations in the APC gene and polyp phenotype. We review the clinical and genetic features of this disorder and provide information on the diagnostic approaches and treatment options available for this disease. PMID- 10805954 TI - Multiple endocrine neoplasias. AB - Multiple endocrine neoplasia type 1 (MEN 1), and the multiple endocrine neoplasia type 2 syndromes (MEN 2A, MEN 2B, and familial non-MEN medullary thyroid carcinoma [FMTC]) encompass a wide range of endocrine problems, but arise from only two genes: the MEN 1 tumor suppressor gene and the RET proto-oncogene. MEN 1 is characterized by parathyroid hyperplasia, pancreaticoduodenal neuroendocrine tumors (PNTs), and pituitary adenomas. Surgery is the principal treatment modality for hyperparathyroidism and PNTs, but questions still remain concerning the timing and extent of surgery for PNTs. The MEN 2 syndromes are characterized by complete penetrance of medullary thyroid cancer. The MEN 2 syndromes differ in their variable expression of hyperparathyroidism, pheochromocytomas, and other clinical features. Genetic testing for mutations in the RET gene has revolutionized treatment by enabling thyroidectomies before significant disease occurs. PMID- 10805955 TI - Psychosocial aspects of cancer genetics: women at high risk for breast and ovarian cancer. AB - In the past five years the advent of cancer genetic testing has created concern about the negative psychosocial sequelae of genetic counseling and testing. Research indicates that the women most likely to seek genetic testing are anxious about carrying a gene mutation and developing breast cancer. Women who are at high risk have poor knowledge and the expectation of being a gene-mutation carrier. High levels of distress have been shown to interfere with decision making about genetic testing. Further, individuals who decline genetic testing may be at increased risk for depressive symptoms even more than those who are found to be gene-mutation carriers. There is great concern that inappropriate candidates will seek genetic testing. Improved education and access to genetic counseling are essential to help women make appropriate decisions about genetic testing. Strategies for the prevention of breast and ovarian cancer are explored, and methods to reduce the adverse psychosocial effects of decision-making about genetic testing and preventive treatment strategies are suggested. PMID- 10805956 TI - Medicolegal and ethical issues in genetic cancer syndromes. AB - Physicians whose patients may be affected by genetic cancer syndromes must be sensitive to a range of legal and ethical concerns. We provide an overview of the major issues, focusing on developments in the United States. The first section examines the physician's legal and ethical duties to the patient. A number of general rules are advanced, including: 1) genetic counseling should be offered to those considering genetic testing; 2) a genetic test should not be performed without the patient's informed consent; and 3) genetic information should not be disclosed to third parties without the patient's written authorization. Laws addressing genetic testing and disclosure of information are reviewed, as well as laws addressing genetic discrimination. Physicians must be aware that the availability of legal protections may affect a patient's willingness to undergo testing. Next, we examine the physician's duty to the patient's family. Where a conflict exists between the interests of the patient and the interests of family members, ethics and current law favor the interests of the patient, absent unusual circumstances. Further, we provide guidance on the scope of the physician's duty to inform, e.g., whether the physician must inform the patient of all medically reasonable treatment options, including those with which the physician disagrees. Finally, we discuss the special ethical and legal issues that prophylactic surgery raises. Areas covered include informed consent and insurance coverage. Several recent cases involving these issues are reviewed. PMID- 10805957 TI - Genetic counseling and clinical cancer genetics services. AB - Cancer genetic services, typically provided by clinicians with expertise in both oncology and genetics, include cancer risk assessment and education, facilitation of genetic testing, pre-and post-test counseling, provision of personally tailored cancer risk management options and recommendations, and psychosocial counseling and support services. All oncology providers should obtain basic information on the family cancer history of their patients to determine the likelihood of hereditary cancer risk as well as possible indications for providing brief or comprehensive cancer genetic counseling. Those who choose to provide these services themselves must be familiar with the complex issues of genetic counseling and testing, and be aware of the time and expertise required to adequately deliver these services. Genetic nurses and genetic counselors with master's degrees function as valuable members of a comprehensive cancer genetic service; they are trained to independently collect and confirm medical and family history information, perform risk assessments, offer patient education regarding cancer and genetics, and provide supportive counseling services for patients and families. It is hoped that specific risk interventions will significantly reduce morbidity and mortality from familial forms of cancer. This review outlines the process of cancer genetic counseling and defines the roles of the cancer genetic counselor and the function of the cancer genetics specialty clinic. The possible medical and legal implications for failing to obtain adequate family history information are reviewed, and the issues of genetic discrimination are discussed. PMID- 10805958 TI - Genetic changes in solid tumors. AB - Although most solid tumors are treated surgically, determining the genetic changes present in the tumor of an individual patient is becoming increasingly important for managing the oncology patient. Our knowledge of the genetic alterations that characterize and predispose to solid tumors continues to expand. Concurrently, the advent of newer technologies such as DNA chips has the potential to enable a more rapid and comprehensive assessment of these changes. The ultimate goal of this new information and technology is to provide sensitive and specific tests that reduce unnecessary procedures and optimize therapy. This review addresses the utility of molecular testing in evaluating cancer. A review of the current technology and hereditary cancer syndromes is also presented. PMID- 10805959 TI - Satellite cell regulation following myotrauma caused by resistance exercise. AB - It is generally accepted that the primary mechanisms governing skeletal muscle hypertrophy are satellite cell activation, proliferation, and differentiation. Specific growth factors and hormones modulate satellite cell activity during normal muscle growth, but as a consequence of resistance exercise additional regulators may stimulate satellite cells to contribute to gains in myofiber size and number. Present knowledge of the regulation of the cellular, biochemical and molecular events accompanying skeletal muscle hypertrophy after resistance exercise is incomplete. We propose that resistance exercise may induce satellite cells to become responsive to cytokines from the immune system and to circulating hormones and growth factors. The purpose of this paper is to review the role of satellite cells and growth factors in skeletal muscle hypertrophy that follows resistance exercise. PMID- 10805961 TI - Effect of serum withdrawal on the proliferation of B16F10 melanoma cells. AB - B16F10 murine melanoma cell proliferation was inhibited after 48 h in medium with serum in the range 0.1 to 0.5% by volume. Cell viability was mostly retained, whereas cells completely deprived of serum died. Growth-arrested cultures showed serum-dependent suppression of DNA synthesis. The response was typically that of a 'cell cycle freeze', verified by flow cytometric distribution of cells. Consequently, serum deprivation did not lead to synchrony when serum was restored to arrested populations. Furthermore, there was no change in PCNA expression in arrested cells. PMID- 10805960 TI - CA(2+) mobilization in the human submandibular duct cell line A253. AB - Ca(2+)mobilization induced by ATP, isoproterenol and the Ca(2+)-ATPase inhibitor thapsigargin in the human submandibular duct cell line A253 was investigated using the Ca(2+)-sensitive fluorescent indicator fura-2. ATP and isoproterenol increased cytosolic free Ca(2+)([Ca(2+)](i)) and subsequent exposure to thapsigargin after ATP or isoproterenol stimulation caused a further increase in [Ca(2+)](i). However, ATP and isoproterenol were not able to elicit a further increase in [Ca(2+)](i)after exposure of the cells to thapsigargin. Relatively few cells reacted to isoproterenol stimulation, but nearly all cells reacted to isoproterenol if ATP was added together with, or prior to isoproterenol stimulation. Moreover, the effect of ATP was potentiated by prior or simultaneous addition of isoproterenol. Furthermore, ATP decreased [Ca(2+)](i)in the presence of thapsigargin probably due to agonist-induced export of intracellular calcium. The results may suggest the existence of three thapsigargin sensitive pools; one opened by ATP acting through P(2)-purinergic receptors and IP(3), one opened by isoproterenol acting through beta2-adrenergic receptors, and a third pool not sensitive to ATP or isoproterenol. PMID- 10805962 TI - Nitric oxide modifies glycolytic pathways in cultured human synoviocytes. AB - Nitric oxide (NO) is a free radical produced during inflammation following activation of an inducible NO synthase by pro-inflammatory cytokines such as IL 1beta. Since both NO and IL-1beta are involved in the physiopathology of inflammatory arthropathies, we investigated the effects of exogenous NO on glycolytic pathways in cultured human osteoarthritic synovial cells. NO generated from S-nitroso-N-acetyl penicillamine (SNAP) or sodium nitroprusside (SNP) inhibited glucose uptake (by 50% after 1 h of incubation) and lactate production by 16% (SNAP) and 8.5% (SNP) after 3 h. Both NO donors also reduced production of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an enzyme of the glycolytic pathway. This effect was reversed by haemoglobin, a NO scavenger with higher affinity for the radical. In contrast, the effect on glucose uptake appeared to be irreversible. PMID- 10805963 TI - Helicobacter pylori vacuolating cytotoxin (VacA) alters cytoskeleton-associated proteins and interferes with re-epithelialization of wounded gastric epithelial monolayers. AB - In previous studies, we demonstrated that Helicobacter pylori vacuolating cytotoxin (VacA) inhibits gastric epithelial cell proliferation and inhibits epidermal growth factor (EGF)-activated signal transduction. Cell proliferation and migration, both essential for mucosal healing are dependent on the cell cytoskeleton. Other investigators demonstrated that VacA induces vacuolation of eukaryotic cells. Since in some cells, control of actin cytoskeleton involves GTP binding proteins of Rho family, in this study we examined whether VacA affects wound re-epithelialization, cell cytoskeleton-associated proteins Rho, Rac1 in a gastric epithelial (RGM1) cell monolayer wound model, and whether these changes correlate with vacuolation. VacA treatment significantly inhibited wound re epithelialization, cell proliferation vs control. VacA-induced cell vacuolation strongly correlated with inhibition of wound re-epithelialization. Furthermore, VacA reduced Rac-1 protein expression and distribution, and C3-mediated ADP ribosylation of Rho. These findings suggest that VacA may interfere with repair of gastric mucosal injury and ulcer re-epithelialization by altering cytoskeleton dependent cell functions and signaling. PMID- 10805964 TI - Evidence of type II estrogen receptor in human osteoblast-like cells. AB - Osteoblast-like cells isolated from human bone bioptic specimens were characterized and analysed for the presence of type II estrogen receptor (type II EBS). The amount of type II EBS was measured by a whole-cell assay at 4 degrees C for 2.5 h using [(3)H]-estradiol as tracer. Saturation analysis, used to investigate the binding characteristic of type II EBS, resulted in a sigmoid curve. Scatchard analysis showed the binding affinity of the estrogen receptor, yielding a concave plot. The dissociation constant (K(d)), determined from the [(3)H]-estradiol concentration required for half saturation was about 12+/-2 nM (SD). The number of type II EBS, estimated at maximum binding, was 197,000+/-8800 sites per cell. If the regulation of the receptor by flavonoids would be confirmed, the evidence of type II EBS in osteoblast-like cells could suggest a direct action of ipriflavone and others flavonoids on bone density in postmenopausal osteoporosis. PMID- 10805965 TI - Expression and purification of biologically active recombinant quail stem cell factor in E. coli. AB - Stem cell factor (SCF) is a multifunctional cytokine involved in hematopoiesis, melanogenesis and gametogenesis. Previous studies have demonstrated that avian SCF is a requirement for the proliferation and survival of various cell types in vivo and in vitro. In the current study, recombinant quail stem cell factor was produced in Escherichia coli using a prokaryotic expression system. SCF was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by affinity chromatography on the Ni-NTA column. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane. Western blot analysis with the monoclonal antibody to the histidine tag identified SCF in the induced cell lysates and the purified sample. The recombinant SCF was approximately 22-23 kD in size. This protein was generated devoid of the signal peptide, the transmembrane domain, and the intracellular domain and, hence, resembles the soluble form of SCF. Biological activity was assayed using the in vitro survival of E12 chicken dorsal root ganglion-derived sensory neurons. The addition of recombinant quail SCF improved neuronal survival. Survival (20.6%) was the highest at the 50 ng/ml concentration of SCF. The availability of quail SCF will be a valuable tool to further resolve the function of stem cell factor in birds. PMID- 10805966 TI - Histochemical characterization of cell death in honeybee larvae midgut after treatment with Paenibacillus larvae, Amitraz and Oxytetracycline. AB - A number of techniques were employed to assess cell death induced in honeybee larvae midgut after per os inoculation of bacterium Paenibacillus larvae var. larvae, the causative agent of American foulbrood disease, and separately with acaricide Amitraz and antibiotic Oxytetracycline. In honeybee larvae exposed to Amitraz, which demonstrates both necrosis and apoptosis, cell death was found in 82% of midgut columnar and in 50% of regenerative epithelial cells, 24 h after treatment. Cell death reduced to 36% in the epithelial cells, 48 h after treatment. In Oxytetracycline-treated larvae, cell death was identified in 40% of midgut epithelial cells, 24 h after inoculation and increased to 55% over the next 24 h. In Paenibacillus -infected larvae, all midgut epithelial cells died. Using ApopTag (Oncor) to label the multiple DNA ends generated by DNA fragmentation showed programmed cell death in 49% of columnar midgut cells 24 h after Amitraz application. Cell death was reduced to 9% over the next 24 h. Our data indicate that cell death could be identified and quantified in situ, using TUNEL techniques. This study also shows that the acaricide Amitraz is a trigger for programmed cell death in the midgut epithelial cells of honeybee larvae, unlike Paenibacillus which induces necrosis only. The data show that immunohistochemical methods are useful for studying in situ tissue pathology, and indicate possibilities for monitoring the effects of infective and chemical environmental stressors on cell death in honeybee larvae tissue. PMID- 10805967 TI - Reduced gut intraepithelial lymphocytes in VLA1 null mice. AB - Very late antigen 1 (VLA1) is an integrin collagen receptor that is expressed by lymphocytes in several disease states. VLA1 blockade has been shown to ameliorate gut disease in experimental graft-versus-host disease. Here we show that in the VLA1 null mouse, which is generally healthy, there is a 50% reduction in gut intraepithelial lymphocytes (IELs) despite an otherwise normal lymphocyte distribution in peripheral blood and lymphoid organs. The gammadelta to alphabeta ratios of IELs are unchanged. We also find that IL2-stimulated splenocytes from VLA1 null animals show a deficiency in adhesion to fibrillar and basement membrane collagen as well as reduced proliferation in response to collagen substratum. These results suggest that some, but not all, intraepithelial lymphocytes require VLA1 to survive or proliferate within the gut epithelium or possibly to traverse the basement membrane. PMID- 10805968 TI - Perillyl alcohol inhibits TCR-mediated [Ca(2+)](i) signaling, alters cell shape and motility, and induces apoptosis in T lymphocytes. AB - Perillyl alcohol (POH) inhibits isoprenylation and has shown anticancer and chemopreventive properties in rodent models. The mechanism that underlies the anticancer activity of POH and other isoprenylation inhibitors is unknown but has been postulated to involve decreased levels of isoprenylated Ras and Ras-related proteins. Previously we demonstrated that POH effectively inhibits human T cell proliferation in vitro and can prevent acute and chronic rejection in a rat cardiac transplant model. In this report, we investigate the effects of POH on T lymphocytes at the single-cell level. POH disrupts the polarized shape and motility of antigen-specific murine 1E5 T cells. Using an optical trap to position anti-CD3-coated beads in contact with 1E5 T cells, we demonstrate that POH inhibits their TCR-mediated calcium response. Furthermore, we show that POH preferentially induces apoptosis in PHA-activated human T cells as well as in 1E5 T cells. PMID- 10805969 TI - Stimulation of thymocyte proliferation by phosphorothioate DNA oligonucleotides. AB - DNA is a complex macromolecule the immunological properties of which depend on short sequence motifs called CpG motifs or immunostimulatory sequences (ISS). These sequences are mitogenic for B cells and can stimulate macrophage cytokine production. While these sequences do not directly activate T cells, they can augment effects of stimulation via the TCR. Furthermore, ISS can affect T cells because of macrophage production of IL-12 and IFN-alpha/beta. In these studies, we further evaluated the immune effects of DNA on T cells, testing the possibility that certain T cell populations can respond directly to this stimulus. We therefore tested the in vitro responses of thymocytes to a series of phosphodiester (Po) and phosphorothioate (Ps) oligonucleotides (ODNs) varying in sequence. In in vitro cultures, phosphorothioate ODNs (sODNs) containing CpG motifs induced significant proliferation of murine thymocytes, although phosphodiester compounds lacked activity. The magnitude of stimulation varied with sequences flanking the CpG motifs, as both dA and dT sequences enhanced the stimulatory capacity of the CpG motif. Furthermore, CpG sODNs were strong costimulators of anti-CD3-mediated thymocyte activation, increasing proliferation compared to anti-CD3 in the absence of DNA. This activation was only partially inhibited by cyclosporine A and was not dependent on a calcium influx. Together, these results indicate that phosphorothioate oligonucleotides containing CpG motifs can directly induce thymocyte proliferation as well as augment TCR activation. These observations thus extend the range of actions of CpG DNA and suggest additional mechanisms for its function as an immunomodulatory agent or adjuvant. PMID- 10805971 TI - Tumor necrosis factor and the p55TNF receptor are required for optimal development of the marginal sinus and for migration of follicular dendritic cell precursors into splenic follicles. AB - The development and function of secondary lymphoid tissue require signaling by tumor necrosis factor and lymphotoxins. Mice deficient in LTbetaR show defective organogenesis of lymph nodes and Peyer's patches and a severely disturbed splenic architecture. In contrast, TNF or p55TNF-R deficiency does not affect the organogenesis of peripheral lymphoid organs but interferes with the formation of B cell follicles and the appearance of FDC networks and germinal centers in all secondary lymphoid organs. Based on these differences, we have previously hypothesized that the role of TNF in lymphoid structure is distinct from that of LT and restricted in regulating cellular interactions that allow the differentiation and/or correct positioning of FDCs. In the present study we show that, in addition to the defects in follicular structure, TNF or p55TNF-R knockout mice exhibit defects in the formation of the macrophage populations and of the sinus lining cells of the splenic marginal zone. Interestingly, a large number of dendritic-shaped cells stained with FDC-specific markers and able to trap immune complexes are retained within the defective marginal zone of TNF and p55TNF-R knockout spleens. We conclude that the primary defect in the lymphoid phenotype of TNF or p55TNF-R knockout mice is the failure of FDC precursors to migrate through the disorganized marginal sinus and to home properly into the splenic follicular areas where they would promote the formation of B cell follicles and germinal centers. PMID- 10805970 TI - Aminopeptidase N/CD13 is directly linked to signal transduction pathways in monocytes. AB - In the present study, we characterized in monocytes the rise in [Ca(2+)](i) evoked by monoclonal antibodies (mAbs) to aminopeptidase N (APN)/CD13, showing a two-phase calcium increase with a small-belled [Ca(2+)](i) rise due to the release of calcium from intracellular stores and a more sustained plateau due to the influx of calcium from the extracellular environment. Tyrosine kinase inhibitors were able to inhibit the rise in [Ca(2+)](i) induced by ligation APN/CD13, as were inhibitors of the phosphatidylinositol 3-kinase. For the first time we can show that mAbs to APN/CD13 provoke phosphorylation of the mitogen activated protein kinases ERK1/2, JNK, and p38. Furthermore, we show that mRNA of the chemotactic cytokine IL-8 is upregulated under the influence of APN/CD13 ligation. Although the in vivo ligand as well as possible cooperating membrane molecules remains to be identified, our results suggest that the membrane ectoenzyme APN/CD13 is a novel signal transduction molecule in monocytes. PMID- 10805972 TI - TGFbeta1 and TNFalpha secreted by mast cells stimulated via the FcepsilonRI activate fibroblasts for high-level production of monocyte chemoattractant protein-1 (MCP-1). AB - Monocytes/macrophages usually make up the largest population of cells in the airways of allergic asthma patients and, as such, contribute substantially to the pathogenesis of this and other allergic diseases. In this report we address one mechanism by which monocytes can be recruited during allergic responses. We and others have shown previously that MCP-1 is important to monocyte infiltration of the tissues during allergic responses in mice and, independently, that mast cells activate fibroblasts to express type alpha1(I)-collagen during such responses. We demonstrate herein that immunologically activated, but not quiescent mouse bone marrow-derived mast cells release mediators which in turn activate primary cultures of embryonic dermal fibroblasts for high-level secretion of monocyte chemoattractant activities. We identify the CC chemokine MCP-1 as a major component of this activity. Anti-MCP-1 antibodies neutralized approximately 80% of the monocyte chemoattractant activities secreted by such mast-cell-activated fibroblasts. Furthermore, our data implicate mast cell TGFbeta and TNFalpha in this process. Depletion of TGFbeta, TNFalpha, or both TGFbeta and TNFalpha from the mediator pool secreted by mast cells activated via the FcepsilonRI reduced the mast-cell-driven fibroblast MCP-1 response by 80+/-15, 56+/-11, or 82+/-5%, respectively. These data thus further delineate a mechanism by which fibroblasts are recruited into and participate in the mast cell-leukocyte cytokine cascades that orchestrate allergic responses. PMID- 10805973 TI - In vivo staphylococcal enterotoxin B (SEB)-primed murine splenocytes secrete mediators which suppress CD25(hi) expression and cell cycle progression of naive splenocytes in response to SEB in vitro. AB - Administration of bacterial superantigen results in clonal activation of T cells followed by a state of hyporesponsiveness to subsequent antigen stimulation. Using a coculture system, we showed that the splenocytes from staphylococcal enterotoxin B (SEB)-injected BALB/c mice suppressed the proliferative response of naive splenocytes to SEB stimulation. The suppressive effect also occurred in Fas deficient MRL-lpr/lpr mice. When naive responder cells were separated by a semipermeable membrane from SEB-primed effector cells, the suppressive effect remained apparent. The hyporesponsiveness of responder cells did not result from excessive induction of apoptosis, but rather from prevention of entering the S and G2/M phases of the cell cycle. The IL-2 levels in culture supernatants were low with the presence of SEB-primed effector cells. However, addition of IL-2 to the cocultures only partially reversed the inhibitory effect. Further studies revealed a reduced level of the CD25(hi) subpopulation in responder cells when cultured in the transwell with the presence of SEB-primed effector cells compared to that with saline-primed controls. This inhibitory effect was not observed for SEB-induced activation of CD25(int) and CD69 expression. Taken together, using a transwell culture system, we show in this study an inhibition of CD25(hi) expression and cell cycle arrest in target cells, which may serve at least in part the mechanisms of SEB-induced hyporesponsiveness. PMID- 10805974 TI - Differential expansion and survival of high and low avidity cytotoxic T cell populations during the immune response to a viral infection. AB - The importance of high avidity CTL for the effective clearance of viral infections is now well established. Thus one would predict that the preferential activation and expansion of high avidity CTL following viral challenge and retention of these cells in the memory pool would be optimal for the immune response. However, whether this actually occurs during the immune response to viral infection is unknown. In this report I have analyzed the avidity of the CTL specific for the OVA(257-264) peptide during acute infection with a recombinant vaccinia expressing ovalbumin and in the memory population. I have found that the relative ratio of high and low avidity CTL varies over the course of an immune response. Thus CTL avidity is an important factor in the expansion and survival of CTL in vivo. PMID- 10805975 TI - Mycobacterium tuberculosis inhibits MHC class II antigen processing in murine bone marrow macrophages. AB - Infection of murine bone-marrow-derived macrophages with viable Mycobacterium tuberculosis (MTB) H37Ra inhibited surface expression of MHC class II (MHC-II) molecules and processing of exogenous antigens for presentation to CD4(+) T hybridoma cells. The inhibition was not dependent on bacterial viability, since it was also produced by exposure to dead bacilli and MTB cytosol preparations, suggesting that it was initiated by a constitutively expressed bacterial component. Northern blot analysis demonstrated that MTB bacilli or cytosol decreased MHC-II mRNA, and immunoprecipitation of biosynthetically labeled molecules confirmed that MHC-II protein synthesis was diminished. Exposure to MTB or MTB cytosol also decreased expression of H2-DM, but H2-DM expression was still sufficient to catalyze conversion of MHC-II to SDS-stable dimers, a measure of MHC-II peptide loading. Thus, infection with MTB decreased both MHC-II and H2-DM expression, but diminished MHC-II synthesis provided the major limitation to antigen processing. PMID- 10805976 TI - Immunohistochemical detection of encephalomyocarditis virus (EMCV) antigen in the heart of experimentally infected piglets. AB - Seven 40-day-old piglets were inoculated with encephalomyocarditis virus (EMCV) strain 424/90, isolated from an outbreak of the myocardial form of the disease in Greece. Two non-infected animals were used as controls. Of the seven inoculated piglets, five died suddenly on day 1.5, 2 (two piglets), 2.5 or 4 post inoculation (p.i. ). The remaining two and the control piglets were killed on day 8 p. i. EMCV antigen was detected immunohistochemically in endothelial cells of capillaries from 1.5 to 2.5 days p.i. only, but was found in cardiac muscle cells, Purkinje fibres and macrophages on all occasions up to day 8 p.i. In endothelial cells and macrophages, EMCV antigen was detected intracytoplasmically, but in cardiac muscle cells and Purkinje fibres it was observed intracytoplasmically or intranuclearly, or both. The frequent presence of EMCV antigen in Purkinje fibres suggests an explanation for the sudden death of the piglets. PMID- 10805977 TI - Canine transmissible venereal tumour: a morphological and immunohistochemical study of 11 tumours in growth phase and during regression after chemotherapy. AB - Eleven dogs with canine transmissible venereal tumour (CTVT) were given vincristine sulphate chemotherapy to induce tumour regression. Biopsy specimens were collected from tumours during the growth phase, before chemotherapy, and again from the same dogs during the regression induced by chemotherapy. Laboratory assessment included cytology, histology, the number of tumour cells in relation to the number of intratumoral leucocytes, proliferative and apoptotic fractions of tumour cells, intratumoral vessel density, and fibrosis. The results revealed that during regression, tumour cell proliferation ceased, apoptosis increased, leucocytes increased (with increased proportion of T lymphocytes), tumour parenchyma collapsed around intratumoral vessels, and fibrosis increased. These results, which were similar to findings in dogs with spontaneous regression of CTVT, suggest that tumour immunity plays a role in tumour regression after modest chemotherapy. PMID- 10805978 TI - Silicate pneumoconiosis in hens. AB - Thirteen cases of silicate pneumoconiosis in 3- to 4-year-old hens are described. Ten of the birds were raised in the suburbs of a city near several chalk quarries and two cement-works; the remaining three hens (aged 3 years) had lived in an environment with high particulate pollution from a nearby brick-works in which large amounts of clay were used daily. Silicotic granulomas composed of dust laden macrophages were scattered over the lungs. They were located mainly in the infundibula and atria of tertiary bronchi and around vessels; more rarely they occurred in the lamina propria mucosae of primary and secondary bronchi. Energy dispersive x-ray microanalysis coupled with both transmission and scanning electron microscopy revealed that the dust was composed mainly of silicon, aluminium, calcium, iron and potassium. Titanium, sulphur, magnesium, zinc, copper and chlorine were also found. It is concluded that animals raised in polluted environmental conditions may serve as an important indicator of risks to human health and pathogenetic mechanisms. Thirteen cases of silicate pneumoconiosis in 3- to 4-year-old hens are described. Ten of the birds were raised in the suburbs of a city near several chalk quarries and two cement-works; the remaining three hens (aged 3 years) had lived in an environment with high particulate pollution from a nearby brick-works in which large amounts of clay were used daily. Silicotic granulomas composed of dust-laden macrophages were scattered over the lungs. They were located mainly in the infundibula and atria of tertiary bronchi and around vessels; more rarely they occurred in the lamina propria mucosae of primary and secondary bronchi. Energy dispersive x-ray microanalysis coupled with both transmission and scanning electron microscopy revealed that the dust was composed mainly of silicon, aluminium, calcium, iron and potassium. Titanium, sulphur, magnesium, zinc, copper and chlorine were also found. It is concluded that animals raised in polluted environmental conditions may serve as an important indicator of risks to human health and pathogenetic mechanisms. PMID- 10805979 TI - Classification of lesions observed in natural cases of paratuberculosis in goats. AB - Sixty-eight adult goats with clinical or sub-clinical paratuberculosis, from naturally infected flocks, were subjected post mortem to a pathological examination that focused on the intestinal lymphoid tissue. The lesions were divided into four categories. Focal lesions, found in 16 goats, consisted of small granulomata in the ileocaecal Peyer's patches or related lamina propria. Diffuse multibacillary lesions (34 goats) consisted of a granulomatous enteritis, affecting different intestinal sites. Numerous macrophages containing many mycobacteria were present, resulting in macroscopical changes in the normal gut morphology; in the ileum of nine of the 34 animals, however, these changes were confined to the apex of the villi and the intestinal wall was not thickened. In diffuse lymphocytic lesions (10 goats), the lymphocyte was the main inflammatory cell, with some macrophages (containing few if any mycobacteria). In diffuse mixed lesions (eight goats) the infiltrate consisted of numerous lymphocytes and macrophages, with small numbers of mycobacteria. The three types of diffuse lesion were often associated with necrosis in the lymph vessels of the mucosa, mesentery and lymph nodes, and with greater thickening of the jejunum than of the ileum. Mycobacterium avium subspecies paratuberculosis was cultured from 69% of goats with diffuse lesions and from 44.4% of those with focal lesions. PMID- 10805980 TI - Genital diseases in the peruvian dusky dolphin (Lagenorhynchus obscurus). AB - Ovarian cysts and one ovarian tumour, uterine tumours, vaginal calculi, abscesses of the broad ligament or undetermined testicular lesions were observed in 25 out of 502 female and male dusky dolphins (Lagenorhynchus obscurus) caught off Peru in 1985-1987 or 1993-1994. Tentative or definitive diagnoses included Graafian follicle cysts, luteinized cyst, ovarian parasitic granulomatous inflammation, dysgerminoma, leiomyoma, fibroleiomyoma and chronic fibrino-suppurative inflammation of the broad ligament. All lesions described represented first reports for L. obscurus, and the diagnosis of dysgerminoma was the first in a cetacean. It is also the first time that trematode eggs have been reported in the ovaries of cetaceans and that a vaginal calculus has been encountered in a sexually immature cetacean. The finding of struvite as a major component in two vaginal calculi suggested an infectious aetiology. Of 11 mature females with ovarian tumour or cysts or uterine tumours, only one (9.1%) was pregnant, i.esignificantly less than the expected pregnancy rate (53.3% in a random sample of Peruvian dusky dolphins). Several females with ovarian or uterine lesions and males with aberrant testes were large animals. It is possible that some of these lesions were associated with normal senescence of the reproductive system. PMID- 10805981 TI - A cytokeratin profile of canine epithelial skin tumours. AB - The reactions of a number of epithelial skin tumours in dogs to a panel of monoclonal antibodies against different human cytokeratins (CKs) were examined immunohistochemically, the purpose being to detect a specific CK profile. CK 6 was present in all epithelial skin tumours with the exception of pilomatrixoma. CK 14 was found in basal cell-derived neoplasias and in sebaceous and perianal gland tumours. CK 10/11 was restricted to spinous cell-derived tumours and CK 8/18 was limited to sweat gland tumours. PMID- 10805982 TI - Virulence of the V592 isolate of equid herpesvirus-1 in ponies. AB - The V592 strain of equid herpesvirus-1 (EHV-1), which was originally isolated from a fetus during an abortion epizootic, has proved to be of low virulence in infection studies. Five Welsh Mountain pony mares and one foal were challenged intranasally or by aerosol with this isolate, and monitored clinically and virologically. All six animals shed virus in nasopharyngeal mucus, and viraemia was recorded from day 7 post-infection (PI). Pathological investigations revealed mild rhinitis and bronchiolitis in the mares, with viral antigen expression in degenerating epithelial cells of the nasal mucosa and bronchioles, and in occasional monocytes in the respiratory tract-associated lymph nodes. Viral antigen expression was not detected in vascular endothelium of the mares, although vasculitis was seen to have affected small numbers of blood vessels in the dorsocaudal lung regions of a mare examined on day 10 PI. In the foal, respiratory lesions of a more localized nature included infection of vascular endothelium and associated vasculitis. The foal also had localized encephalitis affecting the olfactory lobes of the brain, with viral antigen expression in degenerating olfactory neurons and microglia. The data suggest that the relatively low virulence of strain V592 is associated with a lower degree of endotheliotropism than that shown by the highly virulent Ab4 and Army 183 isolates, and that this property is influenced by host immunity. PMID- 10805983 TI - Natural caprine herpesvirus 1 (CpHV-1) infection in kids. AB - Twenty-seven kids aged 5-7 days from a flock of 200 goats in which a high rate of abortion occurred died over a 2-month period. All showed hyperthermia, abdominal pain and anorexia. Two of the kids were examined post mortem. Ulcerative and necrotic lesions affected the whole intestine, and macroscopical changes were also observed in the lungs, urinary bladder and liver. Histologically, a severe necrotizing enteritis as well as thickening of the alveolar septa and necrotic bronchiolo-alveolitis were detected. Prominent microscopical lesions were also present in the liver, urinary bladder, spleen, thymus, mesenteric lymph nodes and kidney. Macrophages containing eosinophilic intranuclear inclusion bodies appeared to be the main inflammatory cell in all the organs examined. Ultrastructurally, herpesvirus particles were evident. Characteristic morphological features of type-A capsids (empty), type-C capsids (large core), and type-B capsids were observed. A virus was isolated in cell culture from all organs examined in the two kids. Cytopathogenic effects (rounding cells, syncytia, vacuoles, cell lysis) and acidophilic intranuclear inclusions typical of herpesvirus infection were observed. The virus was identified as caprine herpesvirus 1 (CpHV-1) by the polymerase chain reaction (PCR) technique and by serum neutralization PMID- 10805984 TI - SHORT PAPER - extensive spinal meningioma in a young dog. AB - The meningioma is one of the most common intradural spinal neoplasms in dogs. Such tumours are usually solitary, well-defined masses that tend to occur in mature and aged dogs. Several variants have been described in the medical and veterinary literature. This paper records a case of a spinal meningioma that extended from the cervical to the lumbosacral spine, with concurrent hydrocephalus, in a 5-month-old male Rhodesian Ridgeback. The tumour appeared to be a meningothelial meningioma with focal mineralization. PMID- 10805985 TI - Epidermolytic ichthyosis in a dog: clinical, histopathological, immunohistochemical and ultrastructural findings. AB - Epidermolytic hyperkeratosis (EH) in man is a complex of congenital, ichthyosiform skin diseases characterized clinically by blistering and hyperkeratosis. These clinical signs are the result of a collapse of the cytoskeleton, seen ultrastructurally as tonofilament clumping and cytolysis within terminally differentiating epidermal cells. In man, specific mutations in keratin 1, 2e, 9 or 10 underlie the various types of EH. This report describes the clinical, histopathological, immunohistochemical and ultrastructural findings in a 6-month-old dog with severe multifocal hyperkeratosis. The morphological changes were comparable with those of EH in man, indicating that this disease, presumably with a similar underlying pathogenetic mechanism, also occurs in the dog. PMID- 10805986 TI - Lymphangiosarcoma in a horse. AB - A 15-year-old Dutch Warmblood mare presented with respiratory distress and anorexia. Clinical examination revealed liquothorax and an abdominal mass. At necropsy, an infiltrating tumour was found in the visceral trunk, continuing through the diaphragm into the mediastinum. Histopathology, immunohistochemistry and electronmicroscopy showed empty channels lined by atypical endothelial cells, and neoplastic cells in a disorganized pattern, consistent with the diagnosis of a lymphangiosarcoma. PMID- 10805987 TI - Uncertainty of the hazardous concentration and fraction affected for normal species sensitivity distributions. AB - Species in the environment vary according to their sensitivity to a toxicant. Because these differences in sensitivity are unique to the toxicant at consideration and laboratory data sets to assess this variability are very small due to cost, it is important to provide uncertainty estimates of (1) environmental quality objectives (hazardous concentrations) derived from these laboratory data and (2) fraction of species affected at given, or predicted, laboratory or environmental concentrations. This article focuses on the normal (Gaussian) distribution of species sensitivity. It examines and compares results of Problems (1) and (2) from two opposing statistical philosophies, Bayesian and Classical, leading to vastly different numerical approaches. For the normal model, both approaches lead to identical answers, numerically. Extrapolation factors for the lower, median, and upper estimates of the hazardous concentration at six levels of protection are derived. Furthermore, upper, median, and lower estimates of the fraction affected at given, standardized, logarithmic concentrations have been tabulated. This table can be used directly for risk assessment without reference to protection levels or hazardous concentrations. The confidence limits for hazardous concentration and fraction affected depend heavily on the number of species tested and are independent of the toxic substance involved (provided the model is right), due to correction for the mean and standard deviation of the toxicity data. The equivalence of confidence limits for hazardous concentration and fraction affected is captured in the law of extrapolation: the upper (median, lower) confidence limit for the fraction affected at the lower (median, upper) confidence limit of the hazardous concentration is equal to the fraction affected (e.g., 5%) used to define the hazardous concentration. The upper confidence limit for the fraction affected at the median estimate of the hazardous concentration for 5% of the species is a fixed number depending on the sample size of the toxicity data only. It amounts to 46% at n=3, down to 20% at n=10, and still 12% at n 30. PMID- 10805988 TI - Studies on hepatic mitochondrial cytochrome P-450 during Plasmodium yoelii infection and pyrimethamine treatment in mice. AB - Hepatic mitochondrial cytochrome P-450 and b(5) activities were significantly depressed, whereas heme and hemozoin were increased during Plasmodium yoelii infection. Type II, aniline-HCl binding efficacy and polyacrylamide gel electrophoretic profile also depicted impairment of cytochrome P-450 during infection. However, the above alterations were more pronounced in the infected hepatic mitochondria, compared to microsomes. Oral treatment of pyrimethamine (10 mg/kg body weightx4 days) to P. yoelii-infected mice brought the altered levels of mitochondrial and microsomal cytochrome P-450 and b(5) to almost normal, whereas heme and hemozoin levels remained unchanged. PMID- 10805989 TI - Coordinated changes of adenylate energy charge and ATP/ADP: use in ecotoxicological studies. AB - The coordinated variations of the adenylate energy charge and ATP/ADP ratio were modeled and a function that depends on the numerical value of the adenylate kinase-catalyzed reaction has been derived. The model allows sensitive detection of the effects of xenobiotics on adenylate kinase and its cellular environment and offers a robust estimation of the direct or indirect effects of pollutants on the adenylate kinase system: data obtained in laboratory studies on shrimp exposed to cadmium and in field studies on oysters either exposed to polychloro biphenyl compounds or located in a heavily polluted area indicate that xenobiotics affect the adenylate kinase reaction directly or by changing its cellular environment. These results demonstrate that application of the model to the treatment of ecotoxicological data allows detection of energetic changes that would have been missed by simple analysis of the usual energetic parameters, and should overcome problems encountered in using energetic parameters during assessment of pollution monitoring. PMID- 10805990 TI - Aluminum fractions in drinking water from reservoirs. AB - To provide insight into aluminum speciation in raw and in finished water and to investigate the factors that can affect it, specific aluminum fractions have been isolated and analyzed in water samples from three drinking water reservoirs in Sardinia (Italy) and at the outlet of their treatment plants. All water treatment plants employed polyaluminum chloride as coagulant. The results demonstrate that the treatment of raw water with the aluminum-based coagulant did not increase the concentration of the metal in the finished water. Aluminum fractions were quite different in raw water and in finished water. More than 80% of the aluminum in raw water was in the particulate form. In the dissolved fraction, organic forms were present at higher concentrations in the raw water, while following water treatment (coagulation, flocculation, filtration) most of the aluminum was in the inorganic form. Most of the dissolved Al in raw water was strongly bound or polymeric colloidal, while percentages of monomeric Al ranging from 40 to 62% were present in the finished water. PMID- 10805991 TI - Nonylphenol affects the granulation pattern of epidermal mucous cells in rainbow trout, Oncorhynchus mykiss. AB - Nonylphenol is a biodegradation product of nonionic surfactants and has recently attracted considerable attention due to its estrogenic potential. Sexually mature male rainbow trout were repeatedly exposed (one to four periods of 10 days each) to environmentally relevant concentrations of nonylphenol (1 microg/L, 10 microg/L) and for comparison, trout were injected with estradiol. Since estrogens are known to induce structural changes within the fish skin, a similar effect of xenobiotics with estrogen-like activity was assumed. Samples of skin were evaluated by means of light and electron microscopy and histochemistry. In trout exposed to nonylphenol and to estradiol, the structure of the epidermis was altered: an irregular overall architecture was often accompanied by detached pavement cells, vacuolation of the cytoplasm, and severely deformed cell nuclei. However, the granulation pattern of the mucous cells was influenced exclusively after exposition to nonylphenol. The number of large and irregularly shaped mucosomes depended more on the exposure period than on the concentration of nonylphenol. Furthermore, this alteration has not yet been reported for any other pollutant or stressor and, thus, can be classified as an effect that would strongly indicate exposure to nonylphenol. PMID- 10805992 TI - Standardized growth toxicity testing (Cu, Zn, Pb, and pentachlorophenol) with Helix aspersa. AB - Juvenile Helix aspersa (1 month, 1 g) were exposed for 4 weeks to food contaminated with copper, zinc, lead, and pentachlorophenol. At concentrations observed in contaminated soils, two essential metals at low levels (Cu and Zn) had a dose-dependent sublethal action on growth. Copper inhibited growth dose dependently between 1000 and 2000 microg small middle dotg(-1) (EC(50)=1200 microg small middle dotg(-1)), whereas zinc had a toxic effect from 4000 microg small middle dotg(-1) (EC(50)=5500 microg small middle dotg(-1)) on. Lead, a nonessential metal, had no negative effect on growth, unlike cadmium (EC(50)=140 microg small middle dotg(-1)), as reported previously. Pentachlorophenol inhibited growth at a concentration of 500 microg small middle dotg(-1) from the fourth week and 1000 microg small middle dotg(-1) from the first week on. The results obtained with these key organisms in the food chain (consumers) complement those obtained with other land invertebrates (earthworms, springtails, wood-lice, etc.). The findings of the present study and those of earlier studies indicate that juvenile snails are useful organisms for testing the sublethal toxicity of chemicals acting via the food, i.e., the main route of toxicant uptake in land animals. PMID- 10805993 TI - Role of glutathione in Thiobencarb resistance in the European eel Anguilla anguilla. AB - Glutathione-dependent defense against xenobiotic toxicity is a multifaceted phenomenon that has been well characterized in mammals. In the present study, eels of species Anguilla anguilla were exposed to 15 ppm of the herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate) for 96 h. Eels exposed to the pesticide were grouped in 24-h intervals according to their time of death, while surviving intoxicated eels constituted another group (live animals). Glutathione content (GSx, GSH, GSSG) was determined in liver and muscle tissues of the dead and live (intoxicated) animals and compared to control values (nonexposed eels). The fish that died before 96 h of exposure were considered susceptible to thiobencarb, while those dead after 96 h and the surviving ones were called resistant. Hepatic glutathione (GSH) content in susceptible eels was lower than that in the control fish, while resistant eels presented GSH levels threefold higher than those of controls. Muscle glutathione levels in dead eels were practically unaffected, but there was a significant decrease in GSH levels in the surviving intoxicated eels. These results indicate that the eels that were able to induce glutathione synthesis in the liver due to the presence of thiobencarb in the medium demonstrated a greater longevity than those who lost glutathione homeostasis. PMID- 10805994 TI - Alterations on AChE activity of the fish Anguilla anguilla as response to herbicide-contaminated water. AB - The inhibition of both total and specific acetylcholinesterase activities was measured in the whole eyes of the yellow eel Anguilla anguilla after exposure to the carbamate thiobencarb. In vivo assays were conducted under a constant flow through system of thiobencarb-contaminated water (1/60 LC(50) 96 h=0.22 ppm for 96 h) followed by a recovery period in clean water (192 h more). The results indicated a measurable level of AChE activity on eyes of control eels, which resulted in a sensitive indicator of the presence of thiobencarb in the water. The pesticide induced significant inhibitory effects on AChE activity ranging from 35% in total AChE activity to 75% in specific AChE activity. Following 1 week of recovery, AChE activity in eel eyes was still different from that of controls. Specific AChE activity remained significantly depressed (35%). On the other hand, protein content in whole eyes of pesticide-treated eels did not exhibit any significant difference from control animals. It has been accepted that a minimum of 20% reduction in brain AChE would be necessary to be indicative of exposure to anticholinergic agents; however, few data are reported about this effect on sense organs. PMID- 10805996 TI - Assessment of the water-extractable genotoxic potential of soil samples from contaminated sites. AB - A screening method for the evaluation of the water-extractable genotoxic potential of soil is proposed. Due to the low sensitivity of genotoxicity test systems, PAD-1 resin was used as solid phase to concentrate less hydrophilic compounds from aqueous soil extracts. Concentrated and nonconcentrated aqueous soil extracts from 19 soil samples were evaluated using three genotoxicity assays: the umu test according to the German standard method with Salmonella typhimurium TA1535/pSK1002, the NM2009 test with S. typhimurium NM2009, and the SOS Chromotest with Escherichia coli PQ37. All tests were performed according to a uniform protocol using microplates. Results indicate that the nonconcentrated water elutriates should be tested because the genotoxic potential of some soil samples is extremely high. Additionally, water elutriates should be concentrated by solid phase extraction to avoid false-negative results. The results of this study show that the genotoxic risk of sites contaminated with PAHs or mineral oils may be underestimated if only nonconcentrated samples are tested. The solid phase extraction method with a concentration factor of 30 for these low soluble compounds is suitable for distinguishing between background genotoxicity of noncontaminated sites and anthropogenic contaminations. PMID- 10805995 TI - Effects of benzimidazole and triazole fungicide use on epigeic species of Collembola in wheat. AB - Foliar sprays of the systemic fungicides carbendazim, propiconazole, and triadimenol were applied in summer to replicated barrier-enclosed and open plots in a field of winter wheat in southern England at dose rates equivalent to label recommendations. Surface-active Collembola (springtails) were sampled from the experimental plots by suction sampling and pitfall trapping before and after the fungicide applications. No consistent effects of the fungicides on collembolan activity were detected using pitfall trapping but suction sampling revealed a transient negative effect of propiconazole and triadimenol on the overall abundance of higher collembolan taxa. Among individual species, however, fungicide effects varied spatially. Fewer significant treatment effects were obtained in enclosed than in open plots and no consistent effects of carbendazim were detected. The relevance of these findings to current fungicide usage strategies in British arable crops, which include the use of complex tank mixes, is discussed. PMID- 10805997 TI - Cholinesterase activity and hematological parameters as biomarkers of sublethal molinate exposure in Anguilla anguilla. AB - Cholinesterase (ChE) activity was measured in plasma, whole blood [using 5,5' dithiobis(2-nitrobenzoic acid) and 2-PDS as chromophores], brain, and whole eyes of Anguilla anguilla exposed to a sublethal concentration of 11.15 mg/L (one third of the 96-h LC(50)) of the carbamate herbicide molinate. ChE activity was evaluated after 6, 24, 48, 72, and 96 h of pesticide exposure. Results indicated that ChE activity in eel tissues decreased as time of exposure increased, especially in eel blood. Eels exposed to molinate were transferred to a pesticide free water for a recovery period of 4 days and ChE activity was also evaluated. Results indicated that ChE activity for those animals with preexposure to the carbamate was still different from the controls even after the recovery period. The use of hematological parameters for assessing the toxicity of molinate in A. anguilla was also studied. The parameters included the measurement of blood proteins, hematocrit, hemoglobin, erythrocytes, and leukocytes. Molinate exposure produced a decrease in all the hematological parameters tested, which was significant only during the recovery period. These results indicate that ChE activity, as well as hematological parameters, may be useful as a diagnostic test for molinate exposure in aquatic organisms. PMID- 10805998 TI - Physiological perturbations in several generations of Daphnia magna straus exposed to diazinon. AB - Daphnia magna was exposed to sublethal diazinon concentrations (0.05, 0.1, 0.5, 0.75, and 1.0 ng/L) for 21 days. The algae Nannochloris oculata (5x10(5) cells/mL) was used to feed the daphnids. Chronic toxicity tests were carried out using neonates of F(1) (first brood) and F(1) (third brood) offspring generations from parentals (F(0)) preexposed to the organophosphate. The effect of diazinon on survival, reproduction, and growth was monitored for the selected daphnid generations. The parameters used to evaluate pesticide effect on reproduction were mean total young per female, mean brood size, time to first reproduction, mean number broods per female, and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to the pesticide. Reproduction as well as survival was significantly reduced when diazinon concentration increased in the medium. This effect was greater in F(1) (first) and F(1) (third) offspring compared to the parental generation (F(0)) daphnids. The intrinsic rate of natural increase (r) decreased with increasing concentrations of diazinon, especially in those animals from the first and third brood. However, the growth of the exposed organisms decreased in the same order of magnitude in all generations tested. The maximum acceptable toxicant concentration (MATC) was calculated for F(0), F(1) (first), and F(1) (third) generations of D. magna exposed to the pesticide using as parameter of evaluation the intrinsic rate of natural increase (r). The interpolation of these results gave MATC values of 0.62 and 0.07 ng/L pesticide for F(0)-F(1) (first) generations and F(1) (third) generation, respectively. Therefore, F(1) (third) generation seems to be more sensitive to diazinon than generations F(0) and F(1) (first). The EC(50) values have been derived for some selected parameters on the generations of D. magna exposed to diazinon. EC(50) values decreased in F(1) (first) and F(1) (third) generations compared to the parental generation F(0). Expanding the reproduction tests to several generations revealed important information on chronic toxicity that could add to an increased cost-effectiveness in the protection of aquatic environments. PMID- 10805999 TI - Spatial distribution of trace metals in sediments from urban streams of Semarang, Central Java, Indonesia. AB - Elevated environmental concentrations of metals are usually associated with the impact of urbanization. The present study is focused on metal contamination in urban sediments. A field survey was carried out to determine the distribution of four metals, i.e., cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn), in the coastal urban area of Semarang, Central Java, Indonesia. Sediment samples were collected from 101 grids of 2x2 km. To map the spatial distribution of these metals, concentrations of each metal were plotted against the corresponding grid coordinate. Cd was below the detection limit (<0.03 microg/g) in all samples, whereas concentrations of Pb, Zn, and Cu fell into a wide range. Frequency distributions of Pb, Zn, and Cu concentrations indicated a similar pattern, in which the major proportion of the sites had a low metal concentration. Some sites, however, had extremely high metal concentrations, Zn up to 1257 microg/g, Pb up to 2666 microg/g, and Cu up to 448 microg/g. The data were used to define background concentrations for sediments in coastal zones of Indonesia ("reference values"). The proposed reference values are 25.6 microg/g, 132.2 microg/g, and 40.7 microg/g, respectively, for Pb, Zn, and Cu. The degree of metal contamination of each individual site was classified according to the calculated value of a combined pollution index, W. Four categories of the degree of metal contamination were proposed, i.e., unpolluted, slightly polluted, polluted, and heavily polluted. Based on this classification, from 101 sites investigated in the greater Semarang area, 51 are unpolluted, 36 slightly polluted, 9 polluted, and 5 heavily polluted. PMID- 10806000 TI - Associations between trace metals in sediment, water, and guppy, Poecilia reticulata (Peters), from urban streams of Semarang, Indonesia. AB - The present study aims to answer the question whether the amount of metal in aquatic biota reflects the concentrations in the sediment and water, and whether the physico-chemical properties of the water and sediment have any influence on the suspected relationship. A study was made of 101 small streams in the city of Semarang, Central Java, Indonesia. Data on fish occurrence in 63 streams of the greater Semarang indicated that the guppies did not avoid the highly polluted sites. No significant difference in body weight between sites was found. Significant differences were found in metal body concentrations (Pb and Zn) between fish collected from sites with different degrees of pollution. A significant declining trend of Pb concentrations with increasing organism size was observed, whereas for two other metals, Zn and Cu, the concentrations did not depend on the body weight. Apparently, body concentrations of these two metals are regulated and maintained at a certain concentration. For the relationships between metal concentrations in water, sediment and fish, water and sediment parameters, and fish dry weight, the presence of significant multiple correlations and bivariate correlation (in 17 of 91 pairs of variables) indicated that, in general, abiotic parameters and body size had no influence on the metal flux from sediment to water, and into fish. Results of partial correlation analyses further suggested that metal concentrations in the sediments were the most important factor governing the metal body concentrations of fish. The present study indicates that the guppy Poecilia reticulata from urban streams is a potential bioindicator for urban metal pollution, especially with respect to their (1) spatial distribution over sites of all pollution regimes and (2) variation in metal accumulation levels reflecting the degree of pollution. PMID- 10806001 TI - Effects of the fungicide copper oxychloride on the growth and reproduction of Eisenia fetida (Oligochaeta). AB - The article describes a laboratory experiment to determine the effect of copper oxychloride on the earthworm Eisenia fetida. Copper oxychloride was used because it is the most commonly used fungicide in South African vineyards but not much is known about its toxicity to earthworms. In an experiment lasting 8 weeks, newly hatched earthworms of the species E. fetida were exposed to copper oxychloride mixed into a urine-free cattle manure substrate. Four groups of 10 worms were used per concentration level (control (4.02), 8.92, 15.92, 39.47, 108.72, 346.85 mg Cu kg substrate(-1)). The following life-history parameters were measured: earthworm growth in consecutive weeks, survival rate, maturation time, cocoon production, reproduction success, total number of hatchlings produced, and incubation time. Earthworm growth and cocoon production were significantly reduced at copper oxychloride exposure concentrations of 8.92 mg kg(-1) and higher. Reproduction success in the 8.92 mg Cu kg substrate(-1) was highest. From an exposure concentration of 15. 92 mg Cu kg substrate(-1) and higher, there was a considerable impact of copper oxychloride on reproduction. This could be seen from a reduced reproduction success, a reduced mean and maximum number of hatchlings per cocoon, and a longer incubation time, indicating a strong effect of low copper oxychloride concentrations on this earthworm species. PMID- 10806003 TI - Papers to appear in environmental research section A PMID- 10806002 TI - Effects of dietary copper on the Australian freshwater crayfish Cherax destructor. AB - Little information is available on the effects of metals in the diet of parastacids, despite the importance of the freshwater crayfish industry worldwide. There have been no published studies on the effect of either dissolved or dietary copper on the Australian freshwater crayfish Cherax destructor. Herein is reported a study on the bioaccumulation of copper through a simple food chain. The copper was first absorbed by the floating aquatic macrophyte Lemna minor to an average concentration of 74 microg/g, before being fed to C. destructor. In the crayfish, the highest copper concentrations were found in the muscle (56.4 1299.6 microg/g), gills (86.8-714.3 microg/g) and hemolymph (14.8-293.5 microg/g). There was no bioaccumulation or bioconcentration of copper from contaminated dietary material. A general linear model indicated that such treatment had no effect on copper concentrations in the crayfish. The results are consistent with the regulation of copper by C. destructor, with the gills being the main site for absorption and depuration of copper to and from the water column. C. destructor does not appear to be sensitive to dietary copper. PMID- 10806004 TI - Platelet glycoprotein IIb/IIIa inhibition during percutaneous coronary revascularization: what more needs to be proven? PMID- 10806005 TI - Androgens and coronary artery disease. A sex-specific effect of sex hormones? PMID- 10806006 TI - Atherosclerosis: a systemic condition that requires a global approach. PMID- 10806007 TI - New light on the Yentl syndrome. PMID- 10806008 TI - Exercise testing, still the method of choice when evaluating patients with chronic stable angina pectoris. PMID- 10806009 TI - Investigation of syncope: increasing the yield and reducing the cost. PMID- 10806010 TI - The impending global epidemic of cardiovascular diseases. PMID- 10806011 TI - Bi-ventricular pacing in congestive cardiac failure. Current experience and future directions. The Imperial College Cardiac Electrophysiology Group. PMID- 10806012 TI - Men with coronary artery disease have lower levels of androgens than men with normal coronary angiograms. AB - AIMS: High androgen levels are presumed by many to explain the male predisposition to coronary artery disease. However, natural androgens inhibit male atherosclerosis(1). Our aim was to determine whether levels of androgens differ between men with and without coronary artery disease. METHODS AND RESULTS: Ninety male subjects (60 with positive, and 30 with negative coronary angiograms) were recruited. Early morning, fasting blood samples were taken from each patient and free, total and bioavailable testosterone, sex hormone binding globulin, oestradiol, and lipids were measured. Bioavailable testosterone was assayed using a modified technique. Free androgen index was calculated. Men with coronary artery disease had significantly lower levels of free testosterone (mean (standard deviation)); 47.95 (13.77) vs 59.87 (26. 05) pmol. l(-1), P=0.027), bioavailable testosterone; 2.55 (0.77) vs 3.26 (1.18) nmol. l(-1), P=0.005 and free androgen index; 37.8 (10. 4) vs 48.47 (18.3), P=0.005, than controls. After controlling for differences in age and body mass index the differences in free androgen index and bioavailable testosterone remained statistically significant (P=0.008 and P=0.013, respectively). CONCLUSION: Men with coronary artery disease have significantly lower levels of androgens than normal controls, challenging the preconception that physiologically high levels of androgens in men account for their increased relative risk for coronary artery disease. PMID- 10806013 TI - Insulin resistance affects endothelium-dependent acetylcholine-induced coronary artery response. AB - AIMS: This study was designed to investigate the relationship between insulin resistance and the acetylcholine-induced endothelium-dependent coronary artery response in patients without angiographically significant atherosclerotic coronary artery disease and to elucidate the pathophysiological significance of insulin resistance in the early stages of coronary atherosclerosis. METHODS AND RESULTS: Insulin resistance was calculated from fasting plasma glucose and insulin concentration using homeostasis model assessment in 40 patients suspected of having ischaemic heart disease, but without angiographic evidence of atherosclerotic coronary artery disease defined as a discrete stenosis or intimal irregularity. They were selected for an acetylcholine provocation test in both left and right coronary arteries. The homeostasis model assessment level was higher in 16 acetylcholine-positive patients than in 24 acetylcholine-negative patients (1.84+/-1.24 vs 0.72+/-0.62, P<0.01). Comparisons of the percentage change in vessel lumen diameter after the acetylcholine test in each of proximal, mid and distal segments of three coronary arteries among the three groups of low (less than 0.7; n=13), intermediate (0.7 to 1.4; n=13), and high homeostasis model assessment level (more than 1.4; n=14) revealed that a higher level resulted in a worse acetylcholine-induced constrictive response in coronary arteries. CONCLUSION: These results suggest that there is an association between high insulin resistance and coronary vascular endothelial cell dysfunction, and that insulin resistance may be an indicator of early stage coronary artery atherosclerosis not detectable by angiography. PMID- 10806014 TI - Prognostic implications of results from exercise testing in patients with chronic stable angina pectoris treated with metoprolol or verapamil. A report from the Angina Prognosis Study In Stockholm (APSIS). AB - AIMS: To evaluate the prognostic implications of results from exercise testing, and of antianginal treatment among patients with chronic stable angina pectoris. MATERIAL AND METHODS: Out of 809 patients in the Angina Prognosis Study In Stockholm (APSIS), 731 (511 men) performed evaluable exercise tests before and after 1 month on double-blind treatment with metoprolol or verapamil. During a median follow-up of 40 months, 32 patients suffered a cardiovascular death and 29 a non-fatal myocardial infarction. RESULTS: Prognostic implications of results from exercise tests were assessed in a multivariate Cox model which included sex, previous myocardial infarction, hypertension and diabetes mellitus. Maximal ST segment depression, especially if >/=2 mm and occurring after exercise, as well as exercise duration independently predicted cardiovascular death. Similar results were obtained for the combined end-point of cardiovascular death+myocardial infarction. Among patients with a positive exercise test at baseline, verapamil reduced the maximal ST-depression more markedly than metoprolol (P<0. 01). However, when the treatment given and treatment effects on ST-segment depression were added to the Cox model, no impact on prognosis could be detected for either cardiovascular death alone or combined with myocardial infarction. Anginal pain carried no prognostic information. CONCLUSION: Marked ST segment depression during and after exercise, and a low exercise capacity independently predicted an adverse outcome in patients with stable angina pectoris, whereas anginal symptoms had no predictive value. Short-term treatment effects on ischaemia did not seem to influence prognosis. Post-exercise ischaemia should be examined carefully when evaluating patients with stable angina pectoris. PMID- 10806015 TI - Gender differences in diagnosis and treatment of coronary artery disease from 1981 to 1997. No evidence for the Yentl syndrome. AB - AIMS: The aim of the present clinical study was to evaluate whether gender related differences existed as regards the extent and localization of coronary artery lesions in patients with angiographically documented coronary artery disease, and whether these angiographic findings would lead to differences in further management. METHODS AND RESULTS: Over a 16-year period (1981-1997) we evaluated 1894 patients (1526 men, 368 women) with angiographically documented coronary artery disease (luminal stenosis >/=60%). For each patient the coronary angiographic results and subsequent revascularization procedures (percutaneous transluminal coronary angioplasty or coronary artery bypass graft surgery) were analysed. The study period was divided into the early angioplasty years (1981 to 1989) and the current angioplasty years (1990-1997). No gender differences in extent and localization of coronary angiographic lesions were observed. In men and women the incidence of single-vessel disease was 42% and 40%, two-vessel disease 27% and 27%, three-vessel disease 26% and 24%, and left main disease 5% and 8%, respectively (P=ns). Localization of disease in men and women was 36% and 39% for the left anterior descending coronary artery, 34% and 32% for the right coronary artery, and 27% and 26% for the left circumflex coronary artery, respectively (P=ns). There was a significant shift from multi-vessel disease towards single-vessel disease in both men and women (both P<0.001). As to subsequent management, a significant gender difference in favour of women was observed (P=0.021). Over time, the number of angioplasty procedures increased significantly from 11.6% to 23.2% for men (P<0.001), and for women from 17.6% to 28.0% (P=0.025), whereas the number of coronary artery bypass procedures decreased in men from 34.9% to 29. 5% (P=0.024) and in women from 42.6% to 30.6% (P=0.019). Referral to angioplasty (n=535) and coronary artery bypass surgery (n=616) in relation to the extent of the disease did not show any gender bias in favour of men. CONCLUSIONS: Our angiographic findings did not show significant gender differences as regards the extent and localization of coronary artery disease in patients with angiographically documented coronary artery disease. More importantly, no substantial evidence could be found for under-referral of women to subsequent therapeutic management. Therefore our study questions the presence of Yentl syndrome in the current era. PMID- 10806016 TI - The role of carotid atherosclerosis in the distinction between ischaemic and non ischaemic cardiomyopathy. AB - AIM: Sometimes ischaemic cardiomyopathy is a result of severe coronary artery disease of an occult course, without typical symptoms or evidence of myocardial infarction. This form of presentation is usually indistinguishable from non ischaemic dilated cardiomyopathy. Carotid bifurcation atherosclerosis and coronary artery disease have been shown to be strongly associated. We prospectively examined the value of extracranial carotid atherosclerosis in the distinction between ischaemic and non-ischaemic aetiology in patients with clinically unexplained cardiomyopathy. METHODS AND RESULTS: Seventy-eight patients with undetermined dilatation and diffuse impairment of the left ventricular contraction were studied within 28 months. They underwent carotid scan and coronary arteriography. Carotid atherosclerosis was found to be very common in ischaemic and rare in non-ischaemic cardiomyopathy. The presence of at least one abnormal carotid finding (intima-media thickness >1 mm, plaques, severe carotid stenosis) was 96% sensitive and 89% specific for ischaemic cardiomyopathy. CONCLUSION: Carotid scanning may be a useful screening and decision making tool in patients with cardiomyopathy of indecisive cause. Patients with carotid atherosclerosis are likely to suffer from severe coronary artery disease. Coronary angiography and subsequent myocardial viability studies, when indicated, could be considered early during their evaluation. In contrast, a negative carotid scan predicts non-ischaemic cardiomyopathy. PMID- 10806017 TI - Low-dose dobutamine responsiveness in idiopathic dilated cardiomyopathy: relation to exercise capacity and clinical outcome. AB - AIMS: To evaluate myocardial contractile reserve using low-dose dobutamine echocardiography in patients with chronic heart failure secondary to idiopathic dilated cardiomyopathy stratified by peak exercise oxygen consumption (VO(2)). METHODS AND RESULTS: Sixty clinically stable patients (56+/-11 years; 45 males) with idiopathic cardiomyopathy and NYHA class I to III symptoms of heart failure were studied and followed-up for 13+/-3 months. All patients underwent cardiopulmonary exercise testing and low-dose dobutamine. The dobutamine infusion protocol consisted of an initial dose of 2.5 micro. kg(-1)per 3 min, increasing by 2.5 micro. kg(-1)per min every 3 min; the maximal dose was 10 micro. kg(-1)per min. The end-systolic volume index, left ventricular ejection fraction and cardiac output were measured at baseline and peak dobutamine dose and their change calculated as ((peak dose value-baseline value)/baseline value]x100. Ten normal subjects with normal left ventricular function and no coronary artery lesions served as a control group to compare low-dose dobutamine results. All analysed echocardiographic variables either at baseline or following dobutamine infusion were significantly lower in patients with chronic heart failure as a whole compared to the control group. When the patients were grouped according to Weber's classification, a statistically significant decrease in percentange changes in end-systolic volume index (rho=-0.77;P<0.0001), left ventricular ejection fraction (rho=-0.72;P<0.0001) and cardiac output (rho=-0. 82;P<0.0001) from class A to class C was observed. The mean percentage decrease in end systolic volume index following the dobutamine infusion was 28.7+/-9% in class A (peak VO(2)>20 ml. kg(-1). min(-1)), 18.6+/-8% in class B (peak VO(2)between 16 and 20 ml. kg. min(-1)), and only 6.4+/-6% in class C (peak VO(2)between 10 and 16 ml. kg(-1). min(-1)) patient groups. At multivariate analysis, only the percentage change in end-systolic volume index was significantly associated with a peak VO(2)<15 ml. kg(-1). min(-1)(P=0.006). During the follow-up, 17 patients had events (15 readmissions for worsening heart failure and two deaths). At multivariate analysis, only the percentage change in end-systolic volume index was significantly associated with the occurrence of events (P=0.003). The area under the receiver operating characteristic curve for percentage change in end systolic volume index was not significantly different from that for peak VO(2)(0. 86+/-0.04 vs 0.80+/-0.06;P:ns). CONCLUSION: This study indicates that in patients with chronic heart failure secondary to idiopathic cardiomyopathy, the cardiac response to low-dose dobutamine, as assessed by echocardiography, is correlated with peak VO(2), an objective and accurate measure of the severity of the disease and clinical outcome. PMID- 10806018 TI - Diagnosing syncope in clinical practice. Implementation of a simplified diagnostic algorithm in a multicentre prospective trial - the OESIL 2 study (Osservatorio Epidemiologico della Sincope nel Lazio) AB - BACKGROUND: In some patients with syncope health care is inappropriate and ineffective. In a recent observational investigation in community hospitals of the Lazio region of Italy (the OESIL study) 54.4% of patients admitted with syncope from the emergency room were discharged without a conclusive diagnosis. AIM OF THE STUDY: A simplified two-step diagnostic algorithm was developed and prospectively implemented in nine community hospitals of the Lazio region of Italy in order to improve the diagnostic performance of clinicians, thereby reducing the number of undiagnosed patients. STUDY POPULATION: The study population included 195 consecutive patients (85 males and 110 females, mean age 62.5 years, range 13-95 years) presenting with a syncopal spell at the emergency room of one of the nine participating hospitals in a 2-month period. RESULTS: The systematic implementation of the proposed diagnostic algorithm resulted in a striking reduction of undiagnosed cases. The percentage of patients discharged without a conclusive diagnosis decreased from 54.4% to 17.5%. Neurally mediated syncope was diagnosed in 35.2% of cases, cardiac syncope in 20.9% and neurological syncope in 13.8%. CONCLUSIONS: The use of specific, simplified diagnostic guidelines and algorithms results in an improvement of overall clinical performance. However, the development of such decision-making aids should carefully consider the local circumstances of daily clinical practice. PMID- 10806020 TI - ESC news and appointments PMID- 10806019 TI - Transcatheter closure of multiple atrial septal defects. Initial results and value of two- and three-dimensional transoesophageal echocardiography. AB - AIMS: To examine the feasibility of transcatheter closure of multiple atrial septal defects using two Amplatzer devices simultaneously and to describe the importance and the role of two- and three-dimensional transoesophageal echocardiography in the selection and closure of such defects. METHODS: Twenty two patients with more than one atrial septal defect underwent an attempt at transcatheter closure of their atrial septal defects at a mean+/-SD age of 30. 8+/-18.6 years (range 3.7-65.9 years) and mean weight of 56.6+/-25.5 kg (range 12.9-99 kg) using two Amplatzer devices implanted simultaneously via two separate delivery systems. During catheterization, two dimensional transoesophageal echocardiography was performed in all but one patient, during and after transcatheter closure, while three dimensional transoesophageal echocardiography was performed in six patients before and after transcatheter closure. RESULTS: Forty-four devices were deployed in all patients to close 45 defects (one patient with three defects closed by two devices). Two dimensional transoesophageal echocardiography was helpful in selection and in guiding correct deployment of the devices. The mean size of the larger defect, as measured by transoesophageal echocardiography was 12.8+/-5.9 mm and the mean size of the smaller defect was 6.6+/-3.0 mm. The mean size of the larger devices was 15+/-7.5 mm, and 8.4+/-3.7 mm for the smaller. Three dimensional transoesophageal echocardiography provided superior imaging and demonstrated the number, shape and the surrounding structures of the atrial septal defects in one single view. The median fluoroscopy time was 28.7 min. Device embolization with successful catheter retrieval occurred in one patient. Forty-four devices were evaluated by colour Doppler transoesophageal echocardiography immediately after the catheterization with a successful closure rate of 97.7%. On follow-up colour Doppler transthoracic echocardiography demonstrated successful closure in 97.5% at 3 months. CONCLUSIONS: The use of more than one Amplatzer septal occluder to close multiple atrial septal defects is safe and effective. The use of two- and three dimensional transoesophageal echocardiography provided useful information for transcatheter closure of multiple atrial septal defects using two devices. Three dimensional transoesophageal echocardiography enhanced our ability to image and understand the spatial relationship of the atrial septal defect anatomy. PMID- 10806021 TI - Theory of the relation between human brain activity (MEG) and hand movements. AB - Earlier research established that spontaneous changes in human sensorimotor coordination are accompanied by qualitative changes in the spatiotemporal dynamics of neural activity measured by multisensor electroencephalography and magnetoencephalography. More recent research has demonstrated that a robust relation exists between brain activity and the movement profile produced. In particular, brain activity has been shown to correlate strongly with movement velocity independent of movement direction and mode of coordination. Using a recently developed field theoretical model of large-scale brain activity itself based on neuroanatomical and neurophysiological constraints we show here how these experimental findings relate to the field theory and how it is possible to reconstruct the movement profile via spatial and temporal integration of the brain signal. There is a unique relation between the quantities in the theory and the experimental data, and fit between the shape of the measured and the reconstructed time series for the movement is remarkably good given that there are no free parameters. PMID- 10806022 TI - The relation between human brain activity and hand movements. PMID- 10806023 TI - Issues in the coordination of human brain activity and motor behavior. PMID- 10806024 TI - Prefrontal cortex involved in higher cognitive functions. Introduction. PMID- 10806025 TI - Distinguishing the functional roles of multiple regions in distributed neural systems for visual working memory. AB - We have investigated the human neural systems for visual working memory using functional magnetic resonance imaging to distinguish sustained activity during memory delays from transient responses related to perceptual and motor operations. These studies have identified six distinct frontal regions that demonstrate sustained activity during memory delays. These regions could be distinguished from brain regions in extrastriate cortex that participate more in perception and from brain regions in medial and lateral frontal cortex that participate more in motor control. Moreover, the working memory regions could be distinguished from each other based on the relative strength of their participation in spatial and face working memory and on the relative strength of sustained activity during memory delays versus transient activity related to stimulus presentation. These results demonstrate that visual working memory performance involves the concerted activity of multiple regions in a widely distributed system. Distinctions between functions, such as perception versus memory maintenance, or spatial versus face working memory, are a matter of the degree of participation of different regions, not the discrete parcellation of different functions to different modules. PMID- 10806026 TI - Activity in ventrolateral and mid-dorsolateral prefrontal cortex during nonspatial visual working memory processing: evidence from functional magnetic resonance imaging. AB - Whole-brain functional magnetic resonance imaging was used to study five healthy human subjects while they performed two nonspatial visual working memory tasks and one control task. In the first memory task, the subjects were required to view a sequence of three pattern stimuli, randomly selected from a familiar set of four stimuli, and then identify which one of three simultaneously presented stimuli was the one that had not been presented in the previous array. In the other task, the subjects were required to observe an identical sequence of three randomly selected pattern stimuli and then to respond by selecting those same stimuli in the order presented. In comparison to a baseline control task, increases in signal intensity were observed, bilaterally, in the mid-dorsolateral frontal cortex and in the right ventrolateral frontal cortex in both memory tasks. When the two tasks were compared directly, however, the first memory task, which had the higher monitoring requirement, yielded significantly greater signal intensity changes in area 9/46 of the right mid-dorsolateral frontal cortex. These results provide further evidence for the precise functional contribution made by the mid-dorsolateral frontal cortex in visual working memory tasks and concur closely with findings in nonhuman primates. PMID- 10806027 TI - The role of prefrontal cortex in sensory memory and motor preparation: an event related fMRI study. AB - Delayed-response tasks are behavioral paradigms in which subjects must remember stimulus attributes across a delay to subsequently perform the appropriate motor response. Quintana and Fuster (1992), reported that there exist subpopulations of neurons in monkey lateral prefrontal cortex (PFC) whose firing rates during the delay are tuned to either sensorial attributes of the stimulus (i.e., involved in sensory memory) or the direction of a postdelay motor response associated with the stimulus (i.e., involved in motor preparation). We studied human subjects with an event-related fMRI method that would allow us to test the hypothesis that there are regions within the PFC that are recruited during both motor preparation and sensory memory. Subjects performed a delayed-response task with two types of trials that either (1) allowed subjects to prepare during a delay period for a specific motor response or (2) required that subjects maintain a sensory attribute (specifically, color) during a delay period for correct performance postdelay. It was assumed that during the delay periods, the delayed-response trials would engage motor preparation while delayed-match trials would engage sensory memory. Behavioral data supported this assumption. Imaging results support the hypothesis that the PFC is involved in both motor preparation and sensory memory. Furthermore, no selectivity (in terms of intensity of neural representation on the spatial scale of the voxel size <5 mm(3)) for motor preparation over sensory memory (or vice-versa) was detected within the PFC. This latter result fails to support a gross anatomical segregation within the PFC with respect to involvement in these two cognitive processes. PMID- 10806028 TI - An fMRI investigation of cortical contributions to spatial and nonspatial visual working memory. AB - The experiments presented in this report were designed to test the hypothesis that visual working memory for spatial stimuli and for object stimuli recruits separate neuronal networks in prefrontal cortex. We acquired BOLD fMRI data from subjects while they compared each serially presented stimulus to the one that had appeared two or three stimuli previously. Three experiments failed to reject the null hypothesis that prefrontal cortical activity associated with spatial working memory performance cannot be dissociated from prefrontal cortical activity associated with nonspatial working memory performance. Polymodal regions of parietal cortex (inferior and superior parietal lobules), as well as cortex surrounding the superior frontal sulcus (and encompassing the frontal eye fields), also demonstrated equivalent levels of activation in the spatial and object conditions. Posterior cortical regions associated with the ventral visual processing stream (portions of lingual, fusiform, and inferior temporal gyri), however, demonstrated greater object than spatial working memory-related activity, particularly when stimuli varied only along spatial or featural dimensions. These experiments, representing fMRI studies of spatial and object working memory in which the testing procedure and the stimuli were identical in the two conditions, suggest that domain-specific visual working memory processing may be mediated by posterior regions associated with domain-specific sensory processing. PMID- 10806029 TI - Working memory for letters, shapes, and locations: fMRI evidence against stimulus based regional organization in human prefrontal cortex. AB - Investigations of working memory (WM) systems in the frontal cortex have revealed two stimulus dimensions along which frontal cortical representations may be functionally organized. One hypothesized dimension dissociates verbal from nonverbal WM processes, dividing left from right frontal regions. The second hypothesized dimension dissociates spatial from nonspatial WM, dividing dorsal from ventral frontal regions. Here we used functional magnetic resonance imaging to probe WM processes associated with three different types of stimuli: letters (verbal and nonspatial), abstract shapes (nonverbal and nonspatial), and locations (nonverbal and spatial). In a series of three experiments using the "n back" WM paradigm, direct statistical comparisons were made between activation patterns in each pairwise combination of the three stimulus types. Across the experiments, no regions that demonstrated responses to WM manipulations were discovered to be unique to any of the three stimulus types. Therefore, no evidence was found to support either a left/right verbal/nonverbal dissociation or a dorsal/ventral spatial/nonspatial dissociation. While this could reflect a limitation of the present behavioral and imaging techniques, other factors that could account for the data are considered, including subjects' strategy selection, encoding of information into WM, and the nature of representational schemes in prefrontal cortex. PMID- 10806030 TI - The prefrontal cortex: no simple matter. PMID- 10806031 TI - Localization of function all over again. PMID- 10806032 TI - Wavelet analysis of dynamic PET data: application to the parametric imaging of benzodiazepine receptor concentration. AB - Receptor density and ligand affinity can be assessed using positron emission tomography (PET). Biological parameters (B(max)('), k(1), k(2), k(on)/V(R), k(off)) are estimated using a compartmental model and a multi-injection protocol. Parametric imaging of the ligand-receptor model has been shown to be of special interest to study certain brain disorders. However, the low signal-to-noise ratio in kinetic curves at the pixel level hampers an adequate estimation of model parameters during the optimization procedure. For this reason, mapping requires a spatial filter, resulting in a loss of resolution. Filtering the kinetic curves in the frequency domain using the Fourier transform is not appropriate, because of difficulties in choosing a correct and efficient cutoff frequency. A wavelet based filter is more appropriate to such tracer kinetics. The purpose of this study is to build up parametric images at the pixel level while conserving the original spatial resolution, using wavelet-based filtering. Data from [(11)C]flumazenil studies, mapping the benzodiazepine receptor density, were used. An invertible discrete wavelet transform was used to calculate the time frequency signals of the time-concentration PET curves on a pixel-by-pixel basis. Kinetic curves observed from large regions of interest in high and low receptor density regions were used to calibrate the threshold of wavelet coefficients. The shrunken wavelet coefficients were then transformed back to the original domain in order to obtain the filtered PET signal. Maps of all binding parameters were obtained at the pixel level with acceptable coefficients of variation of less than 30% for the B(max)(') parameter in most of the gray matter. A strong correlation between model parameter estimates using the usual regions of interest and parametric imaging was observed for all model parameters (r = 0.949 for the parameter B(max)(')). We conclude that wavelet-based filters are useful for building binding parameter maps without loss of the original spatial resolution of the PET scanner. The use of the wavelet-based filtering method can be extended far beyond the multi-injection protocol. It is likely to be also effective for other dynamic PET studies. PMID- 10806033 TI - fMRI evaluation of somatotopic representation in human primary motor cortex. AB - We used fMRI to map foot, elbow, fist, thumb, index finger, and lip movements in 30 healthy subjects. For each movement type confidence intervals of representational sites in the primary motor cortex (M1) were evaluated. In order to improve the precision of their anatomical localization and to optimize the mapping of cortical activation sites, we used both the assessment of locations in the conventional 3D system and a 2D projection method. In addition to the computation of activation maxima of activation clusters within the precentral gyrus, centers of gravity were determined. Both methods showed a high overlap of their representational confidence intervals. The 2D-projection method revealed statistically significant distinct intralimb locations, e.g., elbow versus index finger movements and index finger versus thumb movements. Increased degree of complexity of finger movements resulted in a spread of the somatotopic location toward the arm representation. The 2D-projection method-based fMRI evaluation of limb movements showed high precision and was able to reveal differences in intralimb movement comparisons. fMRI activation revealed a clear somatotopic order of movement representation in M1 and also reflected different degrees of complexity of movement. PMID- 10806034 TI - Principal component analysis and blind separation of sources for optical imaging of intrinsic signals. AB - The analysis of data sets from optical imaging of intrinsic signals requires the separation of signals, which accurately reflect stimulated neuronal activity (mapping signal), from signals related to background activity. Here we show that blind separation of sources by extended spatial decorrelation (ESD) is a powerful method for the extraction of the mapping signal from the total recorded signal. ESD is based on the assumptions (i) that each signal component varies smoothly across space and (ii) that every component has zero cross-correlation functions with the other components. In contrast to the standard analysis of optical imaging data, the proposed method (i) is applicable to nonorthogonal stimulus conditions, (ii) can remove the global signal, blood-vessel patterns, and movement artifacts, (iii) works without ad hoc assumptions about the data structure in the frequency domain, and (iv) provides a confidence measure for the signals (Z score). We first demonstrate on orientation maps from cat and ferret visual cortex, that principal component analysis, which acts as a preprocessing step to ESD, can already remove global signals from image stacks, as long as data stacks for at least two-not necessarily orthogonal-stimulus conditions are available. We then show that the full ESD analysis can further reduce global signal components and-finally-concentrate the mapping signal within a single component both for differential image stacks and for image stacks recorded during presentation of a single stimulus. PMID- 10806035 TI - Toward noninvasive 3-D imaging of the time course of cortical activity: investigation of the depth of the event-related optical signal. AB - The event-related optical signal (EROS) has been recently proposed as a method for studying noninvasively the time course of activity in localized cortical areas (G. Gratton and M. Fabiani, 1998, Psychonomic Bull. Rev. 5: 535-563). Previous data have shown that EROS has very good temporal resolution and can provide detailed surface activity maps. In the present study we investigated whether the depth of the active area can also be estimated. Nine subjects were run in a study in which the eccentricity of the visual stimuli was varied, and EROS was recorded from medial occipital areas using multiple source-detector distances. Seven of the same subjects were also run through a functional magnetic resonance imaging (fMRI) study using the same protocol. The fMRI data indicated that the depth from the head surface to the cortical area activated increased systematically with the eccentricity of the visual stimuli. The EROS recording indicated a response with a latency of 60-80 ms from stimulation. This response varied systematically with eccentricity, so that the greater the eccentricity of the stimuli, the longer the source-detector distance (and thus the depth) at which the EROS effect was observed. The depth of the brain area generating the EROS effect was estimated using a simple algorithm derived from phantom studies on homogeneous media. The average depth estimates for each eccentricity condition obtained with EROS corresponded with those obtained with fMRI, with discrepancies of less than 1 mm. These data demonstrate that multiple source-detector distances can be used to estimate the depth of the cortical areas responsible for the EROS effects. PMID- 10806036 TI - A PET study of visuomotor learning under optical rotation. AB - We measured the regional cerebral blood flow (rCBF) in six healthy volunteers with PET (positron emission tomography) and H(15)(2)O to identify the areas of the human brain involved in sensorimotor learning. The learning task was visually guided reaching with sensorimotor discrepancy caused by optical rotation. PET measurements were performed in the early and late stages of the adaptation to the sensorimotor perturbation. Control measurements were obtained during an eye movement task and a reaching task without optical rotation. The rCBF data of each learning stage were compared to those of both control conditions. During the early stage, rCBF increases were detected in the rostral premotor cortex bilaterally, the posterior part of the left superior parietal lobule (SPL), and the right SPL including the intraparietal sulcus (IPS). During the late stage, rCBF increases were detected in the left caudal premotor area, the left supplementary motor area proper, the left SPL, the right SPL including the IPS, and the right postcentral sulcus extending to the inferior parietal lobule. These results reveal that sensorimotor learning accompanies changes in the recruited cortical areas during different stages of the adaptation, reflecting the different functional roles of each area for different components of adaptation, from learning of new sensorimotor coordination to retention or retrieval of acquired coordination. PMID- 10806037 TI - Event-related potentials correlate with task-dependent glucose metabolism. AB - Cognitive processing is associated with brain electrical activity that is reflected in event-related potentials (ERP). ERP during a target detection task, and regional cerebral glucose metabolism (CMRglc) measured simultaneously, may be influenced by the same neurophysiologic processes. We tested the hypothesis that ERP factors could be directly correlated with CMRglc to derive functional brain maps of brain activity at 120, 160, 200, 280, and 400 ms following stimulus presentation in a target detection task. We controlled for the potential confounding effects of age, sex, and task accuracy, and correlate target-related and nontarget-related ERP separately. Increases and decreases in CMRglc at each time point were identified with statistical parametric mapping (P < 0.001, uncorrected). The 120- and 160-ms maps were the same for target and nontarget processing, while maps for 280 and 400 ms clearly distinguished between targets and nontargets. Extrinsic (early) cognitive processes that depend mainly on stimulus characteristics show less variation based on stimulus meaning (i.e., letter vs shape; target vs nontarget) than later (intrinsic) cognitive processes. These early effects are lateralized to the left hemisphere, for negative ERP factors, and positive ERP-PET correlations are more likely than negative ERP-PET correlations. Thus, brain areas related to task processing impact both ERP and CMRglc measures, suggesting a shared neurophysiologic mechanism for negative ERP factors and increased CMRglc. Direct statistical analysis of these two measures using statistical parametric mapping provides high spatial and temporal resolution in multisubject experiments, while requiring only a single (18)FDG PET scan per subject. PMID- 10806038 TI - Gender difference in premotor activity during active tactile discrimination. AB - To investigate possible gender differences in tactile discrimination tasks, we measured cerebral blood flow of seven men and seven women using positron emission tomography and (15)O water during tactile tasks performed with the right index finger. A nondiscrimination, somatosensory control task activated the left primary sensorimotor cortex and the left parietal operculum extending to the posterior insula without any gender difference. Compared with the control task, discrimination tasks activated the superior and inferior parietal lobules bilaterally, right dorsal premotor cortex, and dorsolateral prefrontal cortex in both genders, consistent with the notion of right hemisphere involvement during exploratory attentional movements. In both genders, symmetric activation of the superior and inferior parietal lobules and asymmetric activation of the right dorsolateral prefrontal cortex were confirmed. The former is consistent with the spatial representation of the tactile input and the latter with the spatial working memory. However, activation of the dorsal premotor cortex was asymmetric in men, whereas it was symmetric in women, the gender difference being statistically significant. This may suggest gender differences in motor programs for exploration in manipulospatial tasks such as tactile discrimination with active touch, possibly by greater interhemispheric interaction through the dorsal premotor cortices in women than in men. PMID- 10806039 TI - Correlation between human personality and neural activity in cerebral cortex. AB - Personality traits are a variance of behavioral patterns among individuals and may reflect a variance of brain activity, but their neurobiological explanation is still a matter of debate. Cloninger proposed three dimensions of personality traits, each of which has strong correlation with activity in a specific central monoaminergic system. Although this theory has been supported by physiological and genetic studies, it is still unclear how these personality parameters are correlated with the activity of the cortical networks which control human behavior. Here we measured the regional cerebral blood flow (rCBF) at rest in 30 normal volunteers who completed the personality inventory of Cloninger. Voxel-by voxel analysis was employed to identify cortical regions where the rCBF showed significant correlation with any of the three personality parameters. Statistically significant correlation was observed in several paralimbic and neocortical regions and was consistent with the assumed monoaminergic influence on neural activity and the distribution of its projections, in each personality dimension. The results suggest that activity in a variety of cortical regions is associated with human personality traits and lend support to Cloninger's theory concerning central monoaminergic influence on human personality traits. PMID- 10806040 TI - Evidence for a refractory period in the hemodynamic response to visual stimuli as measured by MRI. AB - We investigated the effects of paired presentations of visual stimuli upon the evoked hemodynamic response of visual cortex measured by magnetic resonance imaging (MRI). Stimuli were identical 500-ms high-contrast checkerboard patterns, presented singly or with an interpair interval (IPI) of 1, 2, 4, or 6 s (onset-to onset), followed by an intertrial interval of 16-20 s. Images were acquired at 1.5 Tesla using a gradient-echo echoplanar imaging sequence sensitive to blood oxygenation-level dependent (BOLD) contrast. Single checkerboards evoked a hemodynamic response from visual cortex characterized by a rise at 3 s, peak activation at 5 s, and return to baseline by 10 s. We subtracted subjects' single stimulus hemodynamic response from their paired-stimulus responses to isolate the contribution of the second stimulus. If the hemodynamic responses were fully additive, the residual should be a time-shifted replica of the single stimulus response. However, the amplitude of the hemodynamic response to the second checkerboard was smaller, and the peak latency was longer, than for the first. Furthermore, the amplitude decrement was dependent upon IPI, such that the response to the second stimulus at 1 s IPI was only 55% of that to a single stimulus, with recovery to 90% at a 6 s IPI. Peak latency was similarly dependent upon IPI with longer latencies observed for shorter IPIs. These results demonstrate an extended refractory period in the hemodynamic response to visual stimuli consistent with that shown previously for neuronal activity measured electrophysiologically. PMID- 10806041 TI - Chemical heterogeneity of the living human brain: a proton MR spectroscopy study on the effects of sex, age, and brain region. AB - Brain chemistry was compared between 19 male and female normal volunteers in the age group 19-31 years, across six brain regions and nine metabolites using in vivo proton magnetic resonance spectroscopy. The relative concentrations of N acetyl aspartate, choline, glutamate, glutamine, GABA, inositol, glucose, and lactate were measured relative to creatine within 8-cm(3) brain voxels. These measurements were performed in six brain regions: thalamus and cingulate, insula, sensorimotor, dorsolateral prefrontal, and orbital frontal cortices in the left hemisphere. Total metabolite concentration was highest in prefrontal regions (28% higher in orbital frontal cortex and 18.7% higher in dorsolateral prefrontal cortex compared with insula and thalamus, P < 10(-7)). Subjects 25-31 years of age demonstrated a significant increase in total metabolite concentration in the orbital frontal cortex (35%, P < 10(-7)) and sensorimotor cortex (16.7%, P < 10( 5)) compared to those 19-20 years of age. These two brain regions also showed gender dependence, with women demonstrating increased metabolite concentrations compared to men (9% increase in sensorimotor cortex, P < 0.002, and 2.1% in orbital frontal cortex). Most other brain regions showed no gender- or age dependent differences. The results indicate that the living human brain is chemically heterogeneous. The chemical heterogeneity is sex and age dependent and specific for brain region. PMID- 10806042 TI - Statistical sulcal shape comparisons: application to the detection of genetic encoding of the central sulcus shape. AB - Principal Component Analysis allows a quantitative description of shape variability with a restricted number of parameters (or modes) which can be used to quantify the difference between two shapes through the computation of a modal distance. A statistical test can then be applied to this set of measurements in order to detect a statistically significant difference between two groups. We have applied this methodology to highlight evidence of genetic encoding of the shape of neuroanatomical structures. To investigate genetic constraint, we studied if shapes were more similar within 10 pairs of monozygotic twins than within interpairs and compared the results with those obtained from 10 pairs of dizygotic twins. The statistical analysis was performed using a Mantel permutation test. We show, using simulations, that this statistical test applied on modal distances can detect a possible genetic encoding. When applied to real data, this study highlighted genetic constraints on the shape of the central sulcus. We found from 10 pairs of monozygotic twins that the intrapair modal distance of the central sulcus was significantly smaller than the interpair modal distance, for both the left central sulcus (Z = -2.66; P < 0.005) and the right central sulcus (Z = -2.26; P < 0.05). Genetic constraints on the definition of the central sulcus shape were confirmed by applying the same experiment to 10 pairs of normal young individuals (Z = -1.39; Z = -0.63, i.e., values not significant at the P < 0.05 level) and 10 pairs of dizygotic twins (Z = 0.47; Z = 0.03, i.e., values not significant at the P < 0.05 level). PMID- 10806043 TI - Differential expression of egr-1 in osteoarthritic compared to normal adult human articular cartilage. AB - OBJECTIVE: The purpose of this study was to identify genes that are differentially expressed in normal versus osteoarthritic human articular cartilage as either potential novel therapeutic targets or diagnostic markers of this disease. DESIGN: mRNA was isolated from histologically normal and osteoarthritic adult human articular cartilage. The Differential Display technique was employed which identified differentially expressed genes in the normal and diseased tissue. Northern and reverse Northern hybridization were used to confirm the gene expression pattern. Immunohistochemistry and in-situ hybridization were used to localize expression of Egr-1 protein and mRNA respectively in cartilage. RESULTS: A transcription factor, early growth response protein-1 (Egr-1) was found to be down-regulated more than six-fold in multiple human OA cartilage samples when compared to normal tissue. Immunohistochemistry indicated that Egr-1 was expressed throughout normal adult cartilage, in deep-, mid- and superficial-zones. In contrast, in OA cartilage there was expression of Egr-1 mRNA and protein only in the chondrocytes undergoing cloning. CONCLUSIONS: Egr-1 is differentially expressed in OA versus normal cartilage and because of its role in transcriptional activation and repression and regulation of proliferation, differentiation and apoptosis, Egr-1 may play an important role in the pathogenesis of OA. Up-regulation of Egr-1 may therefore provide a novel therapeutic approach for either the prevention or treatment of OA. PMID- 10806044 TI - Influence of aging on the synthesis and morphology of the aggrecans synthesized by differentiated human articular chondrocytes. AB - OBJECTIVE: Synthesis rates of aggrecans by phenotypically stable human articular chondrocytes and the immobilization of these aggrecans in large aggregates were used as variables reflecting the capability of these cells of restoring the extracellular matrix of articular cartilage in vivo in an aging population. DESIGN: Human articular chondrocytes were isolated from articular cartilage obtained from 33 different donors at autopsy. The chondrocytes were cultured in gelled agarose. Synthesis of aggrecans was investigated using Na(2)(35)SO(4)as a radioactive precursor after a 2-week culture period. Electron microscopic study of aggrecan aggregates was done on the macromolecules accumulated over 3 weeks in culture by the chondrocytes obtained from eight other donors with increasing ages. RESULTS: Sulfate incorporation rates into aggrecans correlated inversely with the age of the donor. The value of sulfate incorporation in aggrecans for chondrocytes obtained from mature cartilage of a 20-year-old individual in this system drops to 50% and 25% for chondrocytes obtained from 45- and 69-year-old individuals respectively. Electron microscopic study of aggrecan aggregates showed that the 'de novo' synthesized hyaluronan molecules were fully loaded with aggrecans. Mature human articular cartilage cells were found to synthesize an aggrecan aggregate which carried an average number of 11.7 to 13.1 aggrecans. Cells obtained from immature donors synthesized aggrecan aggregates of which the hyaluronan chain carried twice the amount of aggrecans. These immature human articular cartilage cells were also found to synthesize significant proportions of large aggrecan aggregates with 20 to over 100 aggrecans immobilized on a single hyaluronan chain. The proportions of these large aggrecan aggregates decreased with increasing age of the donors of the chondrocytes. CONCLUSION: The declining aggrecan synthesis rates and the decreased capability of assembling large molecular size aggregates with increasing age in humans illustrates a progressive failure of the repair function of articular cartilage cells in humans. PMID- 10806045 TI - An analysis of 14 molecular markers for monitoring osteoarthritis: segregation of the markers into clusters and distinguishing osteoarthritis at baseline. AB - OBJECTIVE: To investigate the relationships between serum and urinary molecular markers (MM) used to monitor osteoarthritis. DESIGN: Forty osteoarthritis patients had blood and urine collected at baseline and 1, 3, 6 and 12 months later. Specimens from 20 controls were obtained twice at a one month interval. The concentration of 14 different markers was determined at each time point and the data were analyzed by statistical methodology. RESULTS: The markers could be divided by the method of principal components analysis into five clusters of related markers: inflammation markers (C-reactive protein, tumor necrosis receptor type I and tumor necrosis receptor type II, interleukin 6, eosinophilic cationic protein), bone markers (bone sialoprotein, hydroxylysyl pyridinoline, lysyl pyridinoline), putative markers of cartilage anabolism (carboxypropeptide of type II procollagen, hyaluronan, epitope 846) and catabolism (keratan sulfate, cartilage oligomeric matrix protein), and transforming growth factor beta. Three markers (tumor necrosis factor receptor II, cartilage oligomeric matrix protein and epitope 846) from independent clusters discriminated osteoarthritis patients from controls. Inflammation was not a confounding factor in measurement, but a recognizable distinguishing factor in osteoarthritis. CONCLUSIONS: The markers separated into rational groups on the basis of their covariance, a finding with independent biochemical support. The covariance of markers from the same cluster suggests the use of a representative marker from the cluster to reflect changes in osteoarthritis. If multiple markers are being measured within a single cluster, then the use of a weighted cluster 'factor' may be preferable to the separate use of individual markers. PMID- 10806046 TI - Diacerhein and rhein reduce the ICE-induced IL-1beta and IL-18 activation in human osteoarthritic cartilage. AB - OBJECTIVE: IL-1beta plays a fundamental role in osteoarthritis (OA) pathophysiology and cartilage destruction. Targeting the activation mechanism of this cytokine appears to be important as a therapeutic approach. As the interleukin-1 converting enzyme (ICE) is the physiologic modulator of the production of active IL-1beta, we investigated the effect of diacerhein and its active metabolite rhein used in the treatment of OA patients, on the enzyme expression and synthesis on human OA cartilage. Further, we looked at the effect of both drugs on the production of the active form of IL-1beta and IL-18. METHODS: The expression and synthesis of ICE were investigated on human OA cartilage explants using in-situ hybridization and immunohistochemical methods, respectively. The effect of the drugs on ICE OA chondrocytes was also determined by Northern blotting and a specific ELISA assay. Furthermore, the effect of both drugs on the level of active IL-1beta and IL-18 was examined by immunohistochemistry. RESULTS: Data showed that diacerhein and rhein have no true effect on reducing total ICE mRNA by both Northern blotting analysis and in-situ hybridization. A marked and statistically significant decrease was, however, found for protein production. ELISA showed a reduction of 31% (P< 0.04) for diacerhein and 50% (P< 0.02) for rhein. The drugs' immunohistological cell score reduction was similar to data from the ELISA, and a statistical significant reduction of ICE production was found at both superficial and deep zones of the cartilage. IL-1beta and IL-18 were both preferentially produced in chondrocytes of the superficial zone. For each of these cytokines, both drugs demonstrated a statistically significant decrease in this zone. A marked decrease was also noted in the deep zone, but statistical significance was reached only for rhein. CONCLUSION: These results provide a novel regulatory mechanism by which diacerhein and rhein could exert a down-regulation on IL-1's effect on OA cartilage. PMID- 10806047 TI - Simultaneous changes in bone mineral density and articular cartilage in a rabbit meniscectomy model of knee osteoarthrosis. AB - OBJECTIVE: It was hypothesized that increased bone mineral density of the medial proximal tibia would precede or coincide with the development of more severe cartilage changes after meniscectomy. METHODS: In a rabbit knee model, mineral density of subchondral bone and changes of articular cartilage were monitored 13 to 40 weeks after medial meniscectomy or a sham operation. RESULTS: Both procedures resulted in a decrease of bone mineral density, especially of the medial proximal tibia, which persisted up to 40 weeks (P< 0.02-0.0007). Meniscectomy induced cartilage changes typical for osteoarthrosis (P< 0.009), which progressed over time on the posterior aspect of the medial tibial plateau (P< 0.009), which is physiologically covered by the meniscus, but the procedure also induced iatrogenic changes which were located mainly on the anterior aspect of the concerned compartment, and which did not progress or develop to osteoarthrosis. CONCLUSIONS: The data suggest that the cartilage changes after meniscectomy in this animal model are caused by the surgical trauma, subsequent limb misuse, and altered load distribution, and initially associated by a decrease not an increase in bone mineral density of the proximal tibia. Moreover, the cartilage changes progressed without a simultaneous increase of the bone mineral density at corresponding sites. PMID- 10806048 TI - Glucosamine sulfate modulates dysregulated activities of human osteoarthritic chondrocytes in vitro. AB - OBJECTIVE: The efficacy of glucosamine sulfate (GS) in the symptomatic treatment of patients with osteoarthritis (OA) is suggested to be mediated by still unknown effects on the altered OA cartilage. DESIGN: Using human OA chondrocytes in culture, the effects of GS on protein synthesis, caseinase, collagenase, phospholipase A2 (PLA2) and protein kinase C (PKC) activities as well as production of nitric oxide and cyclic AMP were studied in both cells and culture medium. RESULTS: GS significantly reduced PLA2 activity, and more modestly collagenase activity, in the OA chondrocytes in a dose-dependent manner. By contrast, PLA2 and collagenase activity of the culture medium was not modified. No effects on caseinase activity was seen. GS significantly and dose-dependently increased protein synthesis. GS did not modify nitric oxide and cAMP production but significantly increased PKC production. CONCLUSION: GS modified cultured OA chondrocyte metabolism by acting on PKC, cellular PLA2, protein synthesis and possibly collagenase activation. Extrapolation of the effect to the in-vivo situation remains hypothetical but they might represent some possible mechanisms of action of the drug in human. PMID- 10806049 TI - Tumor necrosis factor alpha can contribute to focal loss of cartilage in osteoarthritis. AB - OBJECTIVE; To evaluate the potential for tumor necrosis factor alpha (TNFalpha) induced focal loss of cartilage in osteoarthritic (OA) knee joints. DESIGN: Fresh cartilage from specified regions of OA joints was immunostained for TNF-receptor (R) bearing chondrocytes. Cartilage explants from the same regions were cultured with or without small amounts of TNFalpha and cumulative GAG release into supernatants measured. Concentrations of TNFalpha, p55 and p75 soluble (s) TNF-R in supernatants from cultured OA and non-arthritic (NA) synovium were measured by ELISA. RESULTS: TNF-R bearing chondrocytes were identified in OA cartilage; more specimens contained p55 TNF-R- than p75 TNF-R-bearing chondrocytes and differences in TNF-R distribution were apparent in cartilage from different regions of the same knees. TNFalpha at 5, 1, 0.5 and 0.25 ng/ml (but not 0.1 ng/ml) significantly increased glycosaminoglycans (GAG) release from cartilage explants in a dose-dependent manner. Variation in susceptibility to TNFalpha was observed in explants from different sites. TNFalpha and p75 sTNF-R, but not p55 sTNF-R, concentrations were significantly higher in OA, as compared with NA, supernatants. A significant correlation between TNFalpha and p75 sTNF-R measurements was apparent only in NA supernatants. CONCLUSIONS: Variations in chondrocyte TNF-R expression occur in OA cartilage in vivo. TNFalpha at concentrations produced by OA synovium in vitro, can degrade cartilage matrix. In most OA supernatants sTNF-R concentrations were insufficient to abrogate the effects of TNFalpha. Thus conditions exist in some OA knees for TNFalpha to contribute to focal loss of cartilage. PMID- 10806050 TI - Increased serum levels of non-collagenous matrix proteins (cartilage oligomeric matrix protein and melanoma inhibitory activity) in marathon runners. AB - OBJECTIVE: Marathon runners have an increased risk of developing joint disease. During and after a 42-km run, elevation of multiple cytokines occurs in the blood, reflecting inflammatory processes. We compared this cytokine response with serum levels of cartilage oligomeric matrix protein (COMP) and melanoma inhibitory activity (MIA), two markers for joint metabolism and/or damage. METHODS: Serum from eight endurance-trained runners was collected shortly before the start of a marathon run, after 31 km, 42 km, 2 h after the end, on the first and on the second morning after the run. For comparison, serum was obtained from 35 healthy controls and 80 patients with knee joint injury, rheumatoid arthritis or osteoarthritis. Serum levels of C-reactive protein (CRP), interleukin-1beta (IL-1beta), interleukin-1 receptor antagonist (IL-1RA), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), soluble interleukin-6 receptor (sIL-6R, gp80), soluble tumor necrosis factor receptor II (sTNFRII, p75), COMP and MIA were measured by ELISA. RESULTS: Compared with healthy controls, the runner's baseline serum levels of TNF-alpha, sIL-6R, COMP and MIA were significantly increased. COMP and MIA levels, higher than the upper normal limits of 5 microg/ml and 6 ng/ml respectively, were found in seven and five of eight runners. The elevated levels of COMP were similar to those found in joint injury or osteoarthritis, and the elevated levels of MIA were comparable to those reported in rheumatoid arthritis. During the run, the serum levels of IL-1RA, IL 6, TNF-alpha and COMP rose significantly, and gradually returned to baseline within 24 h. Only modest changes of CRP, sIL-6R, sTNFRII and MIA occurred during the run. Late elevations of CRP and MIA were observed after 24 and 48 h. The correlation analysis suggests associations between COMP, sIL-6R, TNF-alpha, IL 1RA on one hand and sTNFRII, and MIA and CRP on the other hand. CONCLUSIONS: Elevated baseline levels of COMP and MIA might reflect increased joint matrix turnover and/or damage due to prior extreme physical training. During the run, COMP was increasing possibly due to the severe physical strain on joint structures, associated with the early inflammation. After the run, MIA and CRP increased within 24 h, suggesting a correlation with later inflammatory processes. Thus, our data suggest that COMP and MIA are markers for distinct aspects of joint metabolism and/or damage in both disease and sport. PMID- 10806051 TI - Efficacy of etodolac on gait in hip osteoarthritis as assessed by Bessou's locometer: a randomized, crossover, double-blind study versus placebo. Groupe de Recherche sur le Handicap de L'appareil Locomoteur. AB - OBJECTIVES: To assess the efficacy of a single, oral dose of etodolac (300 mg), a nonsteroidal antiinflammatory drug, on gait and pain in patients with unilateral hip osteoarthritis (hOA). DESIGN: Sixteen patients (8 F, 8 M; mean age: 61+/-11.2 years) with painful hOA were included in a randomized, crossover, double-blind study versus placebo. Space and time parameters were assessed using Bessou's locometer and pain was evaluated using the visual analog scale (VAS) at t0 (before taking the drug), t60 (min), t120, and t180 after taking a 300-mg tablet of etodolac. RESULTS: Walking speed was significantly faster only between t0 and t180 under etodolac versus placebo (P< 0.02). Walking speed increased between t0, t60, t120 and t180 with etodolac (P< 0.003), but not with placebo. Stride length increased (P< 0.0001) only on the hOA side, while the time parameters of gait for etodolac- and placebo-treated patients did not differ. VAS values differed significantly at t0 (P< 0.01) between etodolac and placebo groups, but no significant difference was observed at t60, t120 and t180. CONCLUSIONS: Bessou's locometer was able to demonstrate the efficacy of 300 mg of etodolac on gait in hOA. Walking speed was faster 3 h after taking the drug, essentially due to a greater stride length. Pain reduction in the etodolac group contributed to gait improvement. It was concluded that gait performances improved because of less hip pain and thus a greater range of motion after etodolac intake. PMID- 10806053 TI - Quantitative whole-body autoradiography. PMID- 10806052 TI - Molecular cloning and expression patterns of mouse cartilage oligomeric matrix protein gene. AB - OBJECTIVE: To develop transgenic mice harboring mutations in the COMP gene as animal models for pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED), autosomal dominant disorders characterized by early onset osteoarthritis and epiphyseal abnormalities. As a first step in generating a mouse model for COMP mutations, we have cloned the cDNA of mouse COMP and examined its tissue expression pattern. DESIGN: Total mRNA was isolated from the skeletal tissues of newborn C57BL/6j mice and used as a template for oligo(dT) first-strand cDNA synthesis. The cDNA was used for PCR amplification of COMP using three oligonucleotide primer pairs designed from the published rat COMP cDNA sequence. Nested PCR was used to complete the sequence between the amplified fragments. The entire cDNA was sequenced and the expression pattern of the corresponding transcripts examined by Northern hybridizations. RESULTS: A full-length COMP cDNA was isolated. Analysis showed that the entire translated region of the mouse COMP gene is 2268 bp and the derived amino acid sequence shows 90% homology to human COMP. Of eight adult mouse non-cartilage tissues tested, COMP expression was detected only in testis. PMID- 10806054 TI - Technical validation of radioluminography systems. AB - More and more laboratories are forced to document evidently that experimental results were achieved compliant with the regulatory of Good Laboratory Pratice (GLP). Thus, also for the imaging plate technique, which is widely used in whole body autoradiography, it is necessary to provide procedures for formal validation and continuous quality control. Therefore, several European pharmaceutical companies agreed to carry out a project on this subject. This part considers the technical validation. Methods to prove uniform sensitivity and resolution of the technique were developed and are suggested as standard operation procedures and demonstrated, as well, by examples using the isotope (14)C. The high linearity of the detection method is shown by performing a calibration of the image data to the applied radioactivity. PMID- 10806055 TI - Quantitative whole-body autoradiography: recommendations for the standardization of the method. AB - The objective of this article is to identify the best conditions for preparing, handling, and exposing radioactive sections by using the Fujix BAS 2000 system for quantitative radioluminography. Regarding the influence of thickness of sections, thicker sections may allow shorter exposure times due to the increased radioactivity, but on the other hand they take more time for the freeze-drying process, resolution will be reduced, and the recovery of radioactivity will be lower due to increased self-absorption particularly in organs like bones or teeth. The pretreatment of the sections should depend on the method of exposure. Powdering with talcum is the most recommendable method when vacuum packaging the imaging plates and sections. Spraying with Nobecutan is recommended when using the cartridge method. Particularly for low concentrations, the vacuum-contact method should be the method of choice. To avoid a flare effect, the geometrical arrangement of the sections on the imaging plate (IP) should always be vertical to the scanning direction of the laser. An exposure time longer than 10 days is not recommended and the time between the end of exposure and start of scanning should be as short as possible. In order to reduce the background signal, it is necessary to expose the IPs in a shielding box in a cold environment. No positive chemographic effects of sections were found. PMID- 10806056 TI - Sensitivity of radioluminography using (14)C-labeled tracers in whole-body sections of rats. AB - Radioluminography using photostimulated luminescence recently became popular for the quantitation of radioactivity in whole-body sections. In order to estimate the limits of the method, the sensitivity was investigated using routinously prepared blood standards with known amounts of radioactivity. The blood standards underwent the typical steps similar to conventional whole-body autoradiography including freezing, sectioning, and lyophilizing. The limit of detection, defined as 3 times the standard deviation of background values, decreases with exposure time from 18 Bq/g wet tissue for 1 day of exposure to 6 Bq/g for 7 days of exposure. The respective limit of quantitation defined as 10 times the standard deviation of background is 60 to 20 Bq/g. A similar sensitivity is found when the quantitation limit is defined as reproducible measurement with coefficients of variance below 20%: 60 Bq/g can be quantified if the measure area is above 3.5 mm(2). PMID- 10806057 TI - Quantification of tissue self-absorption of weak beta-radiation in lyophilized whole-body sections of rats. AB - Whole-body autoradiography has been widely used in the investigation of the distribution of radiolabeled compounds in animals. The newly introduced radioluminography offers a reliable way of quantifying the radioactivity distribution within whole-body sections. Since the radioactivity is distributed over the entire depth of the section, self-absorption of beta-radiation in tissues is supposed to relevantly affect the detection of radioactivity at the section surface. The self-absorption of radiation energy ((14)C) was investigated in 28 organs/tissues of routinely produced lyophilized rat sections. Nonradioactive whole-body sections with different thickness between 20 and 120 microm were placed between a homogeneous (14)C source and the imaging plates to detect the transmitted radioactivity. The self-absorption was expressed in terms of percentage of transmission of the radioactivity through the sections. Transmission decreased with increasing section thickness, e.g., from 44% (20 microm) to 28% (120 microm) for blood. Comparison of three complete sets of data disclosed intertissue variations of up to about 30% (i.e., +/-15%) disregarding bone. A defined bandwidth of +/-15% around the blood transmission would cover the transmission of almost all tissues. Thus, for most organs radioactivity can be quantified by direct comparison with radioactive blood calibration samples. PMID- 10806058 TI - Quantitative distribution studies in animals: cross-validation of radioluminography versus liquid-scintillation measurement. AB - The results of a cross-validation of the radioluminography (RLG) and liquid scintillation counting (LSC) methods are presented. The methods for the determination of radioactivity concentrations were compared in 16 organs, after administration of (14)C-labeled substances to rats. LSC measurements of two kinds were used as reference methods for RLG: (1) quantitative determination of radioactivity after conventional dissection (interindividual comparison) and (2) quantitative determination of radioactivity in tissue punches taken from the whole-body sections after they had undergone RLG measurement (intraindividual comparison). Blood standards containing known concentrations were used for calibration. For statistical evaluation log-linear regression analysis of paired concentration values and organ-specific 95% confidence intervals of the log transformed RLG/LSC concentration quotients were compared. For most organs, the slopes of the regression lines and the means of the concentration quotients were within the defined equivalence range of 0.80-1.25. Deviations were distinctly smaller in the intraindividual comparison. For some organs, however, it became clear that found concentrations were affected by self-absorption (RLG) and by differences in sample preparation (LSC). In conclusion, quantification with RLG is a reliable and reproducible method with comparable measurement precision and greater accuracy in respect of tissue localization, compared to LSC (dissection). PMID- 10806059 TI - Precision of measurement of tissue concentrations by RLG. AB - Existing investigations about the precision of radioluminography (RLG) are restricted to descriptive analysis of the tissue samples. The aim of the present experiments was to obtain a general prospective statement about the precision that the RLG method can achieve. Several pharmaceutical companies in Europe participated in the experiments. Albino rats of various strains were dosed with various (14)C-labeled compounds. Whole-body sections were produced, and blood calibration scales were set up with standard radioactivity sources of dog or rat blood. Photostimulated luminescence was detected using Fuji imaging plate BAS III. For each organ separately, variability was investigated on each of the levels: rat, section of rat, region within section, and residual, with the help of variance components. The producing company was seen as a fixed factor and adjusted for. A mixed linear model was fitted to the log-transformed data. The variance component (SD estimate) for the residual term gave the desired prospective statement about the achievable precision of the RLG method. Exponential back transformation from the logarithmic to the natural scale transformed the SD estimates to multiplication factors. In total, 29 organs were investigated. The RLG method was comparable in precision to the dissection/combustion method. PMID- 10806060 TI - Quantitative radioluminography of (125)Iodine whole-body autoradiograms. AB - Radioluminography (RLG) was applied for the quantitation of (125)iodine ((125)I) autoradiograms using the bioimaging analyzer BAS 2000. RLG was performed on 50 microm sections of sagittal whole-body sections of rats and (125)I-radiolabeled erythrocyte calibration scales. Linearity, sensitivity, accuracy, and reproducibility were investigated in comparison to direct (125)I radioactivity measurement. RLG is demonstrated to be a simple, reproducible, and precise analytical tool. However, the spatial resolution of quantitative (125)I RLG is limited by the relatively high average free path of gamma-radiation and X rays. PMID- 10806061 TI - Quantitative whole-body radioluminography-future strategy for balance and tissue distribution studies. AB - The present routine to conduct balance and/or tissue dissection distribution studies has now and then been questioned, because the way they are generally conducted does not produce information in proportion to the spending of animal and personnel resources. Usually only total radioactivity is measured and due considerations are not always taken to the metabolic fate of the label. In this study a different strategy is presented-integrating quantitative whole-body radioluminography and different chromatographic methods on extracts of tissue pieces punched from the whole-body sections. In addition to the saving in cost and time, the proposed integrated whole-body radioluminographic/metabolic profile protocol will provide (i) a detailed picture of the distribution of radioactivity at selected dose levels and time points in male, female, and pregnant animals; (ii) the time course of radioactivity in blood/plasma and tissues selected from the images (approximate half-life and AUC); (iii) accumulated urinary and fecal excretion of radioactivity and an estimate of the proportion of radioactive metabolites; (iv) tissue information about the proportion of parent drug versus metabolites of pieces punched from the whole-body sections; and (v) indications of possible tissue binding. PMID- 10806062 TI - An "Eclipse" over the cell nucleus at the turn of the century PMID- 10806063 TI - Review: chromatin structural features and targets that regulate transcription. AB - The nucleosome and chromatin fiber provide the common structural framework for transcriptional control in eukaryotes. The folding of DNA within these structures can both promote and impede transcription dependent on structural context. Importantly, neither the nucleosome nor the chromatin fiber is a static structure. Histone dissociation, histone modification, nucleosome mobility, and assorted allosteric transitions contribute to transcriptional control. Chromatin remodeling is associated with gene activation and repression. Energy-dependent processes mediate the assembly of both activating and repressive proteins into the nucleosomal infrastructure. Recent progress allows the structural consequences of these processes to be visualized at the chromosomal level. DNA and RNA polymerase, SWI/SNF complexes, histone deacetylases, and acetyltransferases are targeted by gene-specific regulators to mediate these structural transitions. The mistargeting of these enzymes contributes to human developmental abnormalities and tumorigenesis. These observations illuminate the roles of chromatin and chromosomal structural biology in human disease. PMID- 10806064 TI - Review: SMCs in the world of chromosome biology- from prokaryotes to higher eukaryotes. AB - The study of higher order chromosome structure and how it is modified through the course of the cell cycle has fascinated geneticists, biochemists, and cell biologists for decades. The results from many diverse technical avenues have converged in the discovery of a large superfamily of chromosome-associated proteins known as SMCs, for structural maintenance of chromosomes, which are predicted to have ATPase activity. Now found in all eukaryotes examined, and numerous prokaryotes as well, SMCs play crucial roles in chromatid cohesion, chromosome condensation, sex chromosome dosage compensation, and DNA recombination repair. In eukaryotes, SMCs exist in five subfamilies, which appear to associate with one another in particular pairs to perform their specific functions. In this review, we summarize current progress examining the roles these proteins, and the complexes they form, play in chromosome metabolism. We also present a twist in the SMC story, with the possibility of one SMC moonlighting in an unpredicted location. PMID- 10806065 TI - Review: dynamic stability of the interphase nucleus in health and disease. AB - Ongoing export of newly synthesized RNAs, as well as control of transcriptional activity, involves dynamic nucleocytoplasmic transport of proteins. Some proteins that shuttle reside primarily in the nucleus while others are concentrated in the cytoplasm. Moreover, some proteins shuttle continuously, while others shuttle only once. A third group is stimulated to relocate either into or out of the nucleus as a result of interruption of shuttling. In addition to these protein specific events, several physiological stimuli have global effects on nucleocytoplasmic transport. In related events, selected proteins move between distinct sites in the nucleoplasm, others enter and leave the nucleolus, and still others transit between the nuclear envelope and cytoplasmic membranes. These multiple dynamic distributions provide numerous opportunities for precise communication between spatially distant sites in the cell. PMID- 10806066 TI - The dynamics of yeast telomeres and silencing proteins through the cell cycle. AB - Genes integrated near the telomeres of budding yeast have a variegated pattern of gene repression that is mediated by the silent information regulatory proteins Sir2p, Sir3p, and Sir4p. Immunolocalization and fluorescence in situ hybridization (FISH) reveal 6-10 perinuclear foci in which silencing proteins and subtelomeric sequences colocalize, suggesting that these are sites of Sir mediated repression. Telomeres lacking subtelomeric repeat elements and the silent mating locus, HML, also localize to the periphery of the nucleus. Conditions that disrupt telomere proximal repression disrupt the focal staining pattern of Sir proteins, but not necessarily the localization of telomeric DNA. To monitor the telomere-associated pools of heterochromatin-binding proteins (Sir and Rap1 proteins) during mitotic cell division, we have performed immunofluorescence and telomeric FISH on populations of yeast cells synchronously traversing the cell cycle. We observe a partial release of Rap1p from telomeres in late G2/M, although telomeres appear to stay clustered during G2-phase and throughout mitosis. A partial release of Sir3p and Sir4p during mitosis also occurs. This is not observed upon HU arrest, although other types of DNA damage cause a dramatic relocalization of Sir and Rap1 proteins. The observed cell cycle dynamics were confirmed by direct epifluorescence of a GFP-Rap1p fusion. Using live GFP fluorescence we show that the diffuse mitotic distribution of GFP-Rap1p is restored to the interphase pattern of foci in early G1-phase. PMID- 10806067 TI - In vivo observation of a nuclear channel-like system: evidence for a distinct interchromosomal domain compartment in interphase cells. AB - We have investigated the interchromosomal domain compartment in living cells by transfecting cDNA coding for Xenopus vimentin, engineered to contain a nuclear localization signal (NLS), coupled to the green fluorescent protein. In human vimentin-free SW13 cells, this chimeric protein was deposited in body-like "dots" both at 37 degrees C, the nonpermissive temperature for assembly of the amphibian vimentin, and 28 degrees C, the optimal temperature for Xenopus vimentin assembly, indicating that the chimeric protein was assembly incompetent. However, when transfected into a subclone stably expressing Xenopus NLS-vimentin (SW13 SC), the chimeric protein incorporated, as a fluorescent tracer, into the structures formed by NLS-vimentin and allowed us to visualize the outgrowth of the vimentin fibers after a temperature shift to 28 degrees C in living cells. In particular, we followed the time-dependent outgrowth of fibers from nuclear dots, first connecting two dots each and with time three and more, eventually generating a spatially restricted fiber system consisting of few loop-like arrays traversing the nucleus. Virtually identical results were obtained when the temperature was lowered only to 30 and 32 degrees C, respectively. An engineered human NLS-vimentin, without need for temperature shift, formed seemingly identical patterns of nuclear fibrils at 37 degrees C in three additionally transfected human cell lines: MCF-7, PLC, and HeLa. When the epithelial cytokeratin pair 8 and 18 was expressed in the nucleus via an engineered NLS in the cytokeratin 18 gene, more network-like, extended filament arrays were generated. Notably, in cotransfection experiments with Xenopus NLS-vimentin, we observed that the formation of these cytokeratin networks at 37 degrees C initiated from dots that nearly entirely colocalized with the aggregated amphibian NLS-vimentin. After a shift to 28 degrees C, extending Xenopus NLS vimentin and cytokeratin filaments frequently followed the same path through the nucleus. These data indicate that interphase cells contain a seemingly equivalent, accessible interchromosomal space. PMID- 10806068 TI - Review: nuclear structure and DNA replication. AB - DNA replication is a highly conserved process among eukaryotes where it occurs within a unique organelle-the nucleus. The importance of this structure is indicated by the fact that assembly of prereplication complexes on cellular chromatin is delayed until mitosis is completed and a nuclear structure has formed. Although nuclear structure is dispensable for DNA replication in vitro, it does appear to play a role in vivo by regulating the concentration of proteins required to initiate DNA replication, by facilitating the assembly or activity of DNA replication forks, and by determining where in the genome initiation of DNA replication occurs. PMID- 10806069 TI - The roles of the MCM, ORC, and Cdc6 proteins in determining the replication competence of chromatin in quiescent cells. AB - Most eukaryotic cell types can withdraw from proliferative cell cycles and remain quiescent for extended periods. Intact nuclei isolated from quiescent murine NIH3T3 cells fail to replicate in vitro when incubated in Xenopus egg extracts, although intact nuclei from proliferating cells replicate well. Permeabilization of the nuclear envelope rescues the ability of quiescent nuclei to replicate in the extract. We show that origin replication complex (ORC), minichromosome maintenance (MCM), and Cdc6 proteins are all present in early quiescent cells. Immunodepletion of Cdc6 or the MCM complex from Xenopus egg extract inhibits replication of permeable, quiescent, but not proliferating, NIH3T3 nuclei. Immunoblotting results demonstrate that mouse homologues of Mcm2, Mcm5, and Cdc6 are displaced from chromatin in quiescent cells. However, this absence of chromatin-bound Cdc6 and MCM proteins from quiescent cells appears not to be due to the absence of ORC subunits as murine homologues of Orc1 and Orc2 remain chromatin-bound in quiescent cells. Surprisingly, intact quiescent nuclei fail to bind exogenously added XCdc6 or to replicate in Xenopus egg extracts immunodepleted of ORC, even though G1- or S-phase nuclei still replicate in these extracts. Our results identify Cdc6 and the MCM complex as essential replication components absent from quiescent chromatin due to nonfunctional chromatin-bound ORC proteins. These results can explain why quiescent mammalian nuclei are unable to replicate in vivo and in Xenopus egg extracts. PMID- 10806070 TI - Ultrastructural characterization of RPA-containing domains in nuclei assembled in Xenopus egg extracts. AB - We describe novel structural domains in in vitro reconstituted Xenopus sperm nuclei, which we term RPA bodies; RPA is the only known marker of these structures. These bodies contain DNA and represent special chromatin domains as seen by transmission electron microscopy. We show that RPA bodies exhibit a similar ultrastructure in nuclei assembled in high-speed supernatant (HSS) of Xenopus egg extract and in nuclei assembled in HSS supplemented with low-speed supernatant (HSS + LSS nuclei). Moreover, RPA bodies are also formed when sperm chromatin containing double-stranded DNA breaks is incubated with HSS of egg extracts. RPA bodies appear to be compartmentalized. By immunoelectron microscopy we show that RPA is preferentially localized at the periphery of the bodies where DNA synthesis also occurs in HSS + LSS nuclei. PMID- 10806071 TI - Atomic force microscopy of the cell nucleus. AB - In mammals and plants, the cell nucleus is organized in dynamic macromolecular domains involved in DNA and RNA metabolism. These domains can be visualized by light and electron microscopy and their composition analyzed by using several cytochemical approaches. They are composed of chromatin or ribonucleoprotein structures as interchromatin and perichromatin fibers and granules, coiled bodies, and nuclear bodies. In plants, DNA arrangement defines chromocentric and reticulated nuclei. We used atomic force microscopy to study the in situ structure of the plant cell nucleus. Samples of the plants Lacandonia schismatica and Ginkgo biloba were prepared as for electron microscopy and unstained semithin sections were mounted on glass slides. For comparison, we also examined entire normal rat kidney cells using the same approach. Samples were scanned with an atomic force microscope working in contact mode. Recognizable images of the nuclear envelope, pores, chromatin, and nucleolus were observed. Reticulated chromatin was observed in L. schismatica. Different textures in the nucleolus of G. biloba were also observed, suggesting the presence of nucleolar subcompartments. The observation of nuclear structure in situ with the atomic force microscope offers a new approach for the analysis of this organelle at high resolution. PMID- 10806072 TI - Defined nuclear changes accompany the reprogramming of the microspore to embryogenesis. AB - The switch of the gametophytic developmental program toward pollen embryogenesis to form a haploid plant represents an important alternative for plant breeding. In the present study, the switch of the gametophytic developmental program toward a sporophytic pathway, "embryogenesis," has been studied in three different plant species, Brassica, tobacco, and pepper. The switch has been induced by stress (heat shock) at the very responsive stage of the microspore, which is the vacuolate period. As a result, the cell nucleus undergoes striking structural changes with regard to late gametophytic development, including alterations of biosynthetic activities and proliferative activity. An enrichment in HSP70 heat shock protein and in the presence of Ntf6-MAP kinase was observed after inductive treatment in the nuclei during early embryogenesis. This apparently reflected the possible roles of these proteins, specifically the protective role of HSP70 for the nuclear machinery, and signal transduction of Ntf6-MAPK for the entry of cells into proliferation. Importantly, the observed nuclear changes were similar in the three species investigated and represented convenient markers for early monitoring of embryogenesis and selection purposes for obtaining double-haploid plants in plant breeding. PMID- 10806073 TI - Review: perinucleolar structures. AB - Several perinucleolar structures have been described in recent years. This review primarily summarizes recent studies regarding two of these structures, the perinucleolar compartment (PNC) and the Sam68 nuclear body (SNB). A number of studies have explored their ultrastructure and molecular components. Despite their different nuclear localizations, PNCs and SNBs share some common characteristics. They both are enriched with RNA binding proteins and nucleic acids and are predominantly localized to the periphery of the nucleolus. They are observed mostly in transformed cells, although prevalence differs among different cell types and cell lines. Their structural integrity is influenced by the transcriptional state of the cell. However, the functions of both the PNC and the SNB remain unknown. In addition to the PNC and SNB, a perinucleolar structure immunolabeled with an antibody to hnRNP L will be discussed. PMID- 10806074 TI - Compartmentalization of RNA processing factors within nuclear speckles. AB - In the mammalian cell nucleus pre-mRNA splicing factors are organized in a speckled pattern. The fluorescence signal within speckles appears homogeneous when cells are immunolabeled with antibodies directed against pre-mRNA splicing factors and examined by fluorescence microscopy. We have reexamined the speckled domains using serial dilutions of antibodies against SR proteins, snRNPs, and a 3' end processing protein by immunofluorescence and confocal laser scanning microscopy. Using higher antibody dilutions, the speckled domains consist of numerous subdomains that are spherical and heterogeneous in size ranging from 0.2 to 0.5 micrometer in diameter. We refer to these subdomains as "subspeckles." Each speckle is composed of 5 to 50 subspeckles and in some cases in actively transcribing cells, strings and loops of subspeckles were observed to extend from the speckled domains. Upon inhibition of RNA polymerase II transcription, the strings and loops of subspeckles were no longer observed. Subspeckles were also not observed in coiled bodies. Using fluorescence in situ hybridization we found subspeckles to be colocalized with transiently expressed beta-tropomyosin RNA transcripts. The compartmentalization into subspeckles may represent an efficient way of organizing these factors for their subsequent transport to transcription/RNA processing sites. PMID- 10806075 TI - Review: movement of mRNA from transcription site to nuclear pores. AB - Pre-mRNA is transcribed primarily from genes located at the interface between chromatin domains and the interchromatin space. After partial or complete processing and complexing with nuclear proteins, the transcripts leave their site of synthesis and travel through the interchromatin space to the nuclear pores for export to the cytoplasm. It is unclear whether transcripts are tethered within the interchromatin space and move toward the nuclear pores using a metabolic energy-requiring, directed mechanism or, alternatively, move randomly by a diffusion-based process. We discuss here recent progress in understanding this step of gene expression, including our experiments tracking the movement of intranuclear poly(A) RNA in living cells. Our results and those of others are most consistent with a model in which newly synthesized mRNAs diffuse throughout the interchromatin space until they randomly encounter and are captured by the export machinery. Because the export machinery appears to preferentially bind transport-competent mRNAs (complexed with the correct complement of nuclear proteins), this diffusion-based model for intranuclear RNA movement potentially allows for a significant level of posttranscriptional control of gene expression. PMID- 10806076 TI - RNA polymerase II in Cajal bodies of amphibian oocytes. AB - Cajal bodies (coiled bodies) are nuclear organelles that contain a variety of components required for transcription and processing of RNA. Cajal bodies in amphibian oocytes are stained by mAb H14, which recognizes the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II when the heptapeptide repeat is phosphorylated on serine-5. Oocytes were treated with the transcription inhibitor 5, 6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB), which prevents phosphorylation of the CTD. Cajal bodies from oocytes that had been treated for 2 3 h with DRB no longer stained with mAb H14, but staining reappeared when the inhibitor was washed out. Epitope-tagged transcripts of two small subunits of polymerase II, RPB6 and RPB9, were injected into the cytoplasm of Xenopus and Triturus oocytes. Newly translated RPB6 and RPB9 were specifically targeted to Cajal bodies within 4 h, and Cajal bodies remained the site of highest concentration of tagged protein during the next 2 days. These data suggest that polymerase subunits pass through the Cajal bodies with a transit time no greater than a few hours. We discuss the possibility that Cajal bodies are sites of assembly or modification of the transcription machinery of the nucleus. PMID- 10806077 TI - Structure and characterization of the murine p80 coilin gene, Coil. AB - Cajal bodies (coiled bodies, CBs) are nuclear organelles of unknown function and are characterized by a wide variety of components including various basal transcription and cell cycle proteins, the nucleolar proteins fibrillarin and Nopp140, numerous small nuclear ribonucleoproteins, the survival motor neuron protein complex, and the marker protein, p80 coilin. To gain insight into the role of p80 coilin in CBs, we have cloned the murine gene Coil and have mapped it to the distal portion of chromosome band 11D. The approximately 2.6-kb transcript is detectable in all tissues analyzed, with the highest levels in brain and testis. Sequence analysis shows that, like its human counterpart, the mouse coilin gene is composed of seven exons and spans nearly 30 kb of genomic DNA. The predicted amino acid sequence reveals two conserved N- and C-terminal domains, and comparison with the Xenopus SPH-1 protein reveals that these three genes are indeed orthologous. These results should facilitate gene disruption experiments aimed at creating a genetic model system to study CBs. PMID- 10806078 TI - Review: properties and assembly mechanisms of ND10, PML bodies, or PODs. AB - Nuclear domain 10 (ND10), also referred to as PML bodies or PODs, are discrete interchromosomal accumulations of several proteins including PML and Sp100. We describe here developments in the visualization of ND10 and the mechanism of ND10 assembly made possible by the identification of proteins that are essential for this process using cell lines that lack individual ND10-associated proteins. PML is critical for the proper localization of all other ND10-associated proteins under physiological conditions. Introducing PML into a PML -/- cell line by transient expression or fusion with PML-producing cells recruited ND10-associated proteins into de novo formed ND10, attesting to its essential nature in ND10 formation. This recruitment includes Daxx, a protein that can bind PML and is highly enriched in condensed chromatin in the absence of PML. The segregation of Daxx from condensed chromatin to ND10 by increased accumulation of Sentrin/SUMO-1 modified PML suggests the presence of a variable equilibrium between these two nuclear sites. These findings identify the basic requirements for ND10 formation and suggest a dynamic mechanism for protein recruitment to these nuclear domains controlled by the SUMO-1 modification state of PML. Additional adapter proteins are suggested to exist by the behavior of Sp100, and Sp100 will provide the basis for their identification. Further information about the dynamic balance of proteins between ND10 and the actual site of functional activity of these proteins will establish whether ND10 function as homeostatic regulators or only in storage of excess proteins destined for turnover. PMID- 10806079 TI - Review: transport of tRNA out of the nucleus-direct channeling to the ribosome? AB - Although tRNA was the first substrate whose export from the nuclei of eukaryotic cells had been shown to be carrier-mediated and active, it has only been in the last 2 years that the first mechanistic details of this nucleocytoplasmic transport pathway have begun to emerge. A member of the importin/karyopherin beta superfamily, Los1p in yeast and Xpo-t in vertebrates, has been shown to export tRNA in cooperation with the small GTPase Ran (Gsp1p) from the nucleus into the cytoplasm, where tRNA becomes available for translation. However, Los1p is not essential for viability in yeast cells, suggesting that alternative tRNA export pathways exist. Recent results show that aminoacylation and a translation factor are also required for efficient nuclear tRNA export. Thus, protein translation and nuclear export of tRNA appear to be coupled processes. PMID- 10806080 TI - Comparative spatial localization of protein-A-tagged and authentic yeast nuclear pore complex proteins by immunogold electron microscopy. AB - The nuclear pore complex (NPC) mediates protein and RNP import in and RNA and RNP export out of the nucleus of eukaryotic cells. Due to its genetic tractability, yeast offers a versatile system for investigating the chemical composition and molecular architecture of the NPC. In this context, protein A tagging is a commonly used tool for characterizing and localizing yeast NPC proteins (nucleoporins). By preembedding anti-protein A immunogold electron microscopy (immunogold EM), we have localized two yeast nucleoporins, Nsp1p and Nic96p, in mutant yeast strains recombinantly expressing these nucleoporins tagged with four (Nsp1p) or two (Nic96p) IgG binding domains of protein A (i.e., ProtA-Nsp1p and ProtA-Nic96p). We have compared the location of the recombinant fusion proteins ProtA-Nsp1p and ProtA-Nic96p (i.e., as specified by their protein A tag) to the location of authentic Nsp1p and Nic96p (i.e., as defined by the epitopes recognized by corresponding nucleoporin antibodies) and found all of them to reside at the same three NPC sites. Hence, recombinant expression and protein A tagging of the nucleoporins Nsp1p and Nic96p have not caused any significant mislocation of the fusion proteins and thus enabled mapping of these two yeast nucleoporins at the ultrastructural level in a faithful manner. PMID- 10806081 TI - Recombinant Nup153 incorporates in vivo into Xenopus oocyte nuclear pore complexes. AB - Nup153 is a molecular constituent of the nuclear basket of the nuclear pore complex (NPC) that plays a critical role in nuclear export of RNAs and proteins. In an effort to map this nucleoporin more precisely within the nuclear basket we have developed an experimental approach for localizing Nup153 expressed and incorporated in vivo into Xenopus oocyte NPCs. This approach involves the microinjection into the cytoplasm of Xenopus oocytes of in vitro synthesized mRNA from a vector encoding an epitope-tagged cDNA. Here we present results obtained by Western blots, fluorescence microscopy, and immuno-electron microscopy, which clearly document that the heterologous protein is properly expressed, targeted, and incorporated into preexisting Xenopus NPCs. This new approach for localizing nucleoporins within the structure of the NPC overcomes limitations of previous techniques and allows for greater specificity and resolution than have been possible with previous methods. PMID- 10806082 TI - Review: nuclear lamins--structural proteins with fundamental functions. AB - The nuclear lamina is located between the inner nuclear membrane and the peripheral chromatin. It is composed of both peripheral and integral membrane proteins, including lamins and lamina-associated proteins. Lamins can interact with one another, with lamina-associated proteins, with nuclear scaffold proteins, and with chromatin. Likewise, most of the lamina-associated proteins are likely to interact directly with chromatin. The nuclear lamina is required for proper cell cycle regulation, chromatin organization, DNA replication, cell differentiation, and apoptosis. Mutations in proteins of the nuclear lamina can disrupt these activities and cause genetic diseases. The structure and assembly of the nuclear lamina proteins and their roles in chromatin organization and cell cycle regulation were recently reviewed. In this review, we discuss the roles of the nuclear lamina in DNA replication and apoptosis and analyze how mutations in nuclear lamina proteins might cause genetic diseases. PMID- 10806083 TI - Review: the dynamics of the nuclear lamins during the cell cycle-- relationship between structure and function. AB - The nuclear lamins are members of the intermediate filament (IF) family of proteins. The lamins have an essential role in maintaining nuclear integrity, as do the other IF family members in the cytoplasm. Also like cytoplasmic IFs, the organization of lamins is dynamic. The lamins are found not only at the nuclear periphery but also in the interior of the nucleus, as distinct nucleoplasmic foci and possibly as a network throughout the nucleus. Nuclear processes such as DNA replication may be organized around these structures. In this review, we discuss changes in the structure and organization of the nuclear lamins during the cell cycle and during cell differentiation. These changes are correlated with changes in nuclear structure and function. For example, the interactions of lamins with chromatin and nuclear envelope components occur very early during nuclear assembly following mitosis. During S-phase, the lamins colocalize with markers of DNA replication, and proper lamin organization must be maintained for replication to proceed. When cells differentiate, the expression pattern of lamin isotypes changes. In addition, changes in lamin organization and expression patterns accompany the nuclear alterations observed in transformed cells. These lamin structures may modulate nuclear function in each of these processes. PMID- 10806084 TI - Review: lamina-associated polypeptide 2 isoforms and related proteins in cell cycle-dependent nuclear structure dynamics. AB - The lamina-associated polypeptide (LAP) 2 family comprises up to six alternatively spliced proteins in mammalian cells and three isoforms in Xenopus. LAP2beta is a type II integral protein of the inner nuclear membrane, which binds to lamin B and the chromosomal protein BAF, and may link the nuclear membrane to the underlying lamina and provide docking sites for chromatin. LAP2alpha shares only the N-terminus with the other isoforms and contains a unique C-terminus. It is a nonmembrane protein associated with the nucleoskeleton and may help to organize higher order chromatin structure by interacting with A-lamins and chromosomes. Recent studies using mutant proteins have just begun to unravel functions of LAP2 isoforms during postmitotic nuclear reassembly. LAP2alpha associates with chromosomes via an alpha-specific domain at early stages of assembly, possibly providing a structural framework for chromosome reorganization. The subsequent interaction of both LAP2alpha and LAP2beta with the chromosomal BAF may stabilize chromatin structure and target membranes to the chromosomes. At later stages LAP2 may regulate the assembly of lamins. LAP2 isoforms have been found to share a homologous approximately 40 amino acid long region, the LEM domain, with nuclear membrane proteins MAN1 and emerin, which has been implicated in Emery-Dreifuss muscular dystrophy. PMID- 10806085 TI - Review: nuclear events in apoptosis. AB - Initial apoptosis research characterized this form of cell death based on distinct nuclear morphology that was subsequently shown to be associated with the appearance of oligonucleosomal DNA fragments. More recent evidence has indicated that apoptosis depends upon a tightly regulated cellular program for its successful initiation and execution. Molecular participants in this program are present in different subcellular compartments, including the plasma membrane, cytosol, mitochondria, and nucleus. The interplay among these compartments and the exchange of specific signaling molecules are critical for the systematic progression of apoptosis. While numerous reports have described a key role for caspase activity in the signaling and executive steps of apoptotic cell death, there are some instances where well-established nuclear changes, characteristic of this form of cell death, can occur independently of caspase activity. Moreover, evidence indicates that certain nuclear events, including chromatin condensation and DNA fragmentation, are controlled separately and depend upon a persistent supply of energy in vivo. In this review, we discuss our current understanding of the role and regulation of nuclear events in the apoptotic process with an emphasis on protease and endonuclease activities as well as the ability of certain Bcl-2 family proteins to influence this process. PMID- 10806086 TI - Structure of apo-glyceraldehyde-3-phosphate dehydrogenase from Palinurus versicolor. AB - d-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) shows cooperative properties for binding coenzymes. The structure of apo-GAPDH from Palinurus versicolor has been solved at 2.0 A resolution by X-ray crystallography. The final model gives a crystallographic R factor of 0.178 in the resolution range 8 to 2 A. The structural comparison with holo-GAPDH from the same species reveals a conformational change induced by coenzyme binding similar to that observed in Bacillus stearothermophilus GAPDH but to a lesser extent. The differences in magnitude during the apo-holo transition between these two enzymes were analyzed with respect to the change of the amino acid composition in the coenzyme binding pocket. In the crystalline state of apo-GAPDH, the overall structures of the subunits are similar to each other; however, significant differences in temperature factors and minor differences in domain rotation upon coenzyme binding were observed for different subunits. These structural features are discussed in relation to the environmental asymmetry of crystallographically independent subunits. PMID- 10806087 TI - The cell surface glycoprotein layer of the extreme halophile Halobacterium salinarum and its relation to Haloferax volcanii: cryo-electron tomography of freeze-substituted cells and projection studies of negatively stained envelopes. AB - We have studied the surface layer (S-layer) of Halobacterium salinarum (formerly Halobacterium halobium), an extreme halophile requiring high concentrations of sodium, by electron microscopy of (a) isolated, negatively stained, flattened envelopes and (b) cryo-fixation of intact cells in their high-salt growth medium followed by freeze substitution and tomography of thin sections. From the negatively stained isolated envelopes we have calculated a two-dimensional, projection map that is strikingly similar to that of Haloferax volcanii, an extreme halophile requiring high concentrations of magnesium; both projection maps show the hexagonal arrangement of the morphological units with an identical center-to-center spacing of 150 A; each of the morphological units of the two species has six subunits with a similar density distribution and apparent domain organization. In contrast to the two-dimensional map, the tomographic reconstruction of Halob. salinarum does not agree in a straightforward way with the three-dimensional, electron crystallographic map of negatively stained Halof. volcanii envelopes, although the main features of the lattice and the morphological units are evident. The tomographic reconstruction of sections from epoxy-embedded material suffers from directional compression due to sectioning stress and continuous dimensional changes and mass loss due to electron irradiation. This communication consists, therefore, of three parts: (a) a comparison of the projection maps of negatively stained envelopes of Halof. volcanii and Halob. salinarum; (b) a comparison of the three-dimensional maps obtained by electron crystallography (Halof. volcanii) and low-dose cryo tomography (Halob. salinarum); and (c) a methodological study of mass loss and dimensional changes of plastic-embedded material under low-dose conditions at room and liquid nitrogen temperatures. PMID- 10806088 TI - The single mitochondrion of tachyzoites of Toxoplasma gondii. AB - The infective tachyzoite form of the protozoan Toxoplasma gondii is able to penetrate into vertebrate host cells and to survive and multiply within a cytoplasmic vacuole known as the parasitophorous vacuole. Previous observations, confirmed in the present study, showed that extracellular, but not intravacuolar, tachyzoites are labeled with rhodamine 123, a dye that specifically binds to functional mitochondria, which present a high transmembrane potential. These observations led to the suggestion that intravacuolar tachyzoites do not possess functional mitochondria. However, our present observations using the new dye CMXRos and observation by confocal laser scanning microscopy (CLSM) showed that the mitochondria of both extracellular and intravacuolar tachyzoites were intensely labeled, indicating that they were functional. In addition, cytochrome c activity could be cytochemically detected in the inner mitochondrial membrane of intravacuolar tachyzoites. Three-dimensional reconstruction of serial optical sections of CMXRos-stained tachyzoites observed by CLSM and of serial thin sections examined by transmission electron microscopy revealed that the protozoan presented only one ramified mitochondrion, reinforcing previous observations by Seeber et al. (1998, Exp. Parasitol. 89, 137-139) Petitprez and Vivier (1972, Protistologica VIII, 199-221). PMID- 10806089 TI - Plasmodium falciparum-infected erythrocytes: qualitative and quantitative analyses of parasite-induced knobs by atomic force microscopy. AB - We used the combination of an atomic force microscope and a light microscope equipped with epifluorescence to serially image Plasmodium falciparum-infected erythrocytes. This procedure allowed us to determine unambiguously the presence and developmental stage of the malaria parasite as well as the number and size of knobs in singly, doubly, and triply infected erythrocytes. Knobs are not present during the ring stage of a malaria infection but a lesion resulting from invasion by a merozoite is clearly visible on the erythrocyte surface. This lesion is visible into the late trophozoite stage of infection. Knobs begin to form during the early trophozoite stage of infection and have a single-unit structure. Our data suggest the possibility that a two-unit structure of knobs, which was reported by Aikawa et al. (1996, Exp. Parasitol. 84, 339-343) using atomic force microscopy, appears to be a double-tipped image. The number of knobs per unit of host cell surface area is directly proportional to parasite number in both early and late trophozoite stages. These results indicate that knob formation by one parasite does not influence knob formation by other parasites in a multiply infected erythrocyte. In addition, knob volume is not influenced by either parasite stage or number at the late trophozoite stage, indicating that the number of component molecules per knob is constant throughout the parasite maturation process. PMID- 10806090 TI - Polymorphism in the packing of aquaporin-1 tetramers in 2-D crystals. AB - Hitherto, the packing arrangement of the aquaporin-1 (AQP1) tetramer in 2 dimensional (2-D) crystals (two-sided plane group p42(1)2) was observed to be largely similar (canonical crystal form) despite the difference in the source of the protein, the glycosylation state of the protein, the type of lipids, and the ratio of lipid to protein in the crystallization mixture. We report here our observation that the packing of AQP1 tetramers shows polymorphism in 2-D crystals generated in dioleoyl phosphatidylcholine bilayers. Apart from the canonical form, three additional allomorphs were identified. One was observed when small (0.25) lipid to protein ratio was used in the crystallization mixture while the other two were observed when the divalent cation content in the canonical crystals was modified. The various allomorphs were distinguished by different relative orientations of the AQP1 tetramer viewed in projection. The same, two sided plane group p42(1)2 and similar unit cell dimensions were maintained in the different allomorphs as established by analysis of images of frozen-hydrated, nominally untilted crystals. Our results indicate that the interaction between the AQP1 monomers at the interface of the tetramers is flexible and is also strongly influenced by Mg(2+) ions with the cation effect materializing because of the intrinsic fluidity of the membrane. PMID- 10806091 TI - Membrane-bound 3D structures reveal the intrinsic flexibility of annexin VI. AB - Several quasi-ordered arrays and three two-dimensional crystal forms of annexin VI were obtained on artificial lipid monolayers. Three-dimensional reconstructions of the crystal forms exhibit marked differences in the orientations of the two lobes, revealing flexibility of the linker between the two lobes of annexin VI. Evidence is presented that the lobes may bind the monolayer in a parallel orientation, or an antiparallel orientation, in which the second lobe is turned away from the monolayer. It is hypothesized that annexin VI may also adopt several conformations in vivo, underlying different functional roles. PMID- 10806092 TI - Cryopreparation provides new insight into the effects of brefeldin A on the structure of the HepG2 Golgi apparatus. AB - High-pressure freezing and freeze-substitution were used to study Golgi ultrastructure and its brefeldin A-induced transformations in HepG2 human hepatoma cells. Cryoimmobilization arrested subcellular dynamics within milliseconds, thus considerably improving the temporal resolution in monitoring the very early effects of high brefeldin concentrations at the ultrastructural level (i.e., 20 microg/ml brefeldin applied for 35 s to 8 min). Moreover, this approach ruled out possible cumulative and/or synergistic effects of the drug and fixatives. Several findings differed from studies based on chemical fixation. In particular, Golgi breakdown did not proceed gradually but occurred in distinct steps. We found a conspicuous lag between the absence of nonclathrin coats on Golgi membranes after 30 s of brefeldin treatment and the disassembly of the stacks, which did not start until after 90 to 120 s. At this time, domains at the trans and cis faces separated from the stacks, starting tubulation and fragmentation. After 3-5 min the Golgi apparatus was completely replaced by loose meshworks of straight tubules of different sizes and staining properties; also frequent were bent tubules and vesicles forming glomerule-like structures. After 8 min all kinds of Golgi-derived structures had aggregated within huge clusters. The morphologically highly distinct structures found after brefeldin treatment could in part be correlated with particular Golgi domains in the control cells. PMID- 10806093 TI - X-Ray diffraction analysis of three-dimensional crystals of bovine rhodopsin obtained from mixed micelles. AB - Rhodopsin, a prototypic G protein-coupled receptor responsible for absorption of photons in retinal rod photoreceptor cells, was selectively extracted from bovine rod outer segment membranes, employing mixed micelles of nonyl beta-d-glucoside and heptanetriol. Highly purified rhodopsin was crystallized from solutions containing varying amounts of detergent and amphiphile. The crystals contained ground state rhodopsin molecules as judged by their red color and the linear dichroism originating from the 11-cis-retinal chromophore. However, when exposed to visible light, even at 4 degrees C, rhodopsin was bleached and the crystals decomposed. Reflections in the diffraction pattern were observed out to 3.5-A resolution at 100 K for the most ordered crystals. Diffraction data have been processed to 3.85-A resolution. The symmetry of the diffraction pattern and the systematic absences indicate that the crystals have tetragonal symmetry, space group P4(1)22 or P4(3)22, a = b = 96.51 A, c = 148.55 A. A value of 4.12 A(3)/Da for V(M) was obtained for one monomer in the asymmetric unit (eight molecules per unit cell). Our study is the first characterization of a three-dimensional crystal of a G protein-coupled receptor and may be valuable for future structural studies on related receptors of this important superfamily. PMID- 10806094 TI - Crystallization of restriction endonuclease BamHI with nonspecific DNA. AB - Restriction endonucleases show extraordinary specificity in distinguishing specific from nonspecific DNA sequences. A single basepair change within the recognition sequence results in over a million-fold loss in activity. To understand the basis of this sequence discrimination, it is just as important to study the nonspecific complex as the specific complex. We describe here the crystallization of restriction endonuclease BamHI with several nonspecific oligonucleotides. The 11-mer, 5'-ATGAATCCATA-3', yielded cocrystals with BamHI, in the presence of low salt, that diffracted to 1.9 A with synchrotron radiation. The cocrystals belong to the space group P2(1)2(1)2(1) with unit cell dimensions of a = 114.8 A, b = 91.1 A, c = 66.4 A, alpha = 90 degrees, beta = 90 degrees, gamma = 90 degrees. This success in the cocrystallization of BamHI with a nonspecific DNA provides insights for future attempts at crystallization of other nonspecific DNA-protein complexes. PMID- 10806095 TI - Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function. AB - Gene expression is a fundamental cellular process. The basic mechanisms involved in expression of genes have been characterized at the molecular level. A major challenge is now to uncover how transcription, RNA processing and RNA export are organized within the cell nucleus, how these processes are coordinated with each other and how nuclear architecture influences gene expression and regulation. A significant contribution has come from cell biological approaches, which combine molecular techniques with microscopy methods. These studies have revealed that the mammalian cell nucleus is a complex but highly organized organelle, which contains numerous subcompartments. I discuss here how two essential nuclear processes - transcription and pre-mRNA splicing - are spatially organized and coordinated in vivo, and how this organization might contribute to the control of gene expression. The dynamic nature of nuclear proteins and compartments indicates a high degree of plasticity in the cellular organization of nuclear functions. The cellular organization of transcription and splicing suggest that the morphology of nuclear compartments is largely determined by the activities of the nucleus. PMID- 10806096 TI - The Shank family of scaffold proteins. AB - Shank proteins make up a new family of scaffold proteins recently identified through their interaction with a variety of membrane and cytoplasmic proteins. Shank polypeptides contain multiple sites for protein-protein interaction, including ankyrin repeats, an SH3 domain, a PDZ domain, a long proline-rich region, and a SAM domain. Binding partners for most of these domains have been identified: for instance, the PDZ domain of Shank proteins interacts with GKAP (a postsynaptic-density protein) as well as several G-protein-coupled receptors. The specific localization of Shank proteins at postsynaptic sites of brain excitatory synapses suggests a role for this family of proteins in the organization of cytoskeletal/ signaling complexes at specialized cell junctions. PMID- 10806097 TI - Proteolytic processing and cell biological functions of the amyloid precursor protein. AB - Recent research has identified some key players involved in the proteolytic processing of amyloid precursor protein (APP) to amyloid beta-peptide, the principal component of the amyloid plaques in Alzheimer patients. Interesting parallels exists with the proteolysis of other proteins involved in cell differentiation, cholesterol homeostasis and stress responses. Since the cytoplasmic domain of APP is anchored to a complex protein network that might function in axonal elongation, dendritic arborisation and neuronal cell migration, the proteolysis of APP might be critically involved in intracellular signalling events. PMID- 10806098 TI - Transmembrane-4-superfamily proteins CD151 and CD81 associate with alpha 3 beta 1 integrin, and selectively contribute to alpha 3 beta 1-dependent neurite outgrowth. AB - Proteins in the transmembrane-4-superfamily (TM4SF) form many different complexes with proteins in the integrin family, but the functional utility of these complexes has not yet been demonstrated. Here we show that TM4SF proteins CD151, CD81, and CD63 co-distribute with alpha3beta1 integrin on neurites and growth cones of human NT2N cells. Also, stable CD151-alpha3beta1 and CD81-alpha3beta1 complexes were recovered in NT2N detergent lysates. Total NT2N neurite outgrowth on laminin-5 (a ligand for alpha3beta1 integrin) was strongly inhibited by anti CD151 and -CD81 antibodies either together ( approximately 85% inhibition) or alone ( approximately 45% inhibition). Notably, these antibodies had no inhibitory effect on NT2N neurites formed on laminin-1 or fibronectin, when alpha3beta1integrin was not engaged. Neurite number, length, and rate of extension were all affected by anti-TM4SF antibodies. In summary: (1) these substrate-dependent inhibition results strongly suggest that CD151 and CD81 associations with alpha3beta1 are functionally relevant, (2) TM4SF proteins CD151 and CD81 make a strong positive contribution toward neurite number, length, and rate of outgrowth, and (3) NT2N cells, a well-established model of immature central nervous system neurons, can be a powerful system for studies of integrin function in neurite outgrowth and growth cone motility. PMID- 10806099 TI - N-terminal targeting of guanine nucleotide exchange factors (GEF) for ADP ribosylation factors (ARF) to the Golgi. AB - B2-1 (cytohesin-1) is a member of a group of proteins (including ARNO and ARNO3) that are all of similar size and domain composition. The three proteins contain an N-terminal coiled-coil domain, followed by a Sec7 and a pleckstrin homology (PH) domain. While it is well established that the Sec7 domain functions as a guanine nucleotide exchange factor (GEF) for ADP-ribosylation factors (ARFs) and the PH domain anchors the proteins to membrane phosphoinositols, the function of the N-terminal domain is unknown. Here we show that the N terminus of B2-1 (residues 1-54) is necessary and sufficient to target the protein to the Golgi. The Sec7+PH domains of B2-1 (residues 55-398) are not sufficient for Golgi localization. Further deletion analysis and point mutagenesis indicate that the coiled-coil domain within the N terminus is responsible for Golgi targeting. Furthermore, ARNO and ARNO3 N termini also have the same capability of targeting to the Golgi. We conclude that the N-terminal, (&agr;)-helical, coiled-coil domain is used to target this family of proteins to the Golgi complex. PMID- 10806100 TI - Patching plasma membrane disruptions with cytoplasmic membrane. AB - Vesicle-vesicle fusion initiated in cell cytoplasm by high Ca(2+) can rapidly erect large membrane boundaries. These might be used as a 'patch' for resealing plasma membrane disruptions. Three central predictions of this 'patch' hypothesis are here established in sea urchin eggs. First, we show that surface markers for plasma membrane protein and lipid are initially absent over disruption sites after resealing is complete. Second, we demonstrate that resealing capacity is strongly dependent upon local availability of fusion competent cytoplasmic organelles, specifically the reserve or yolk granule. Lastly, we demonstrate that the reserve granule is capable of rapid (t(1/2) <1 second), Ca(2+)-regulated (high threshold) fusion capable of erecting large (>1000 microm(2)), continuous membrane boundaries. Production of patch vesicles for resealing may proceed by an 'emergency' fusion mechanism distinct from that utilized for the much slower, highly regulated, cytosol-requiring organelle-organelle fusion events typical of constitutive membrane trafficking pathways. PMID- 10806101 TI - Centromere clustering is a major determinant of yeast interphase nuclear organization. AB - During interphase in the budding yeast, Saccharomyces cerevisiae, centromeres are clustered near one pole of the nucleus as a rosette with the spindle pole body at its hub. Opposite to the centromeric pole is the nucleolus. Chromosome arms extend outwards from the centromeric pole and are preferentially directed towards the opposite pole. Centromere clustering is reduced by the ndc10 mutation, which affects a kinetochore protein, and by the microtubule poison nocodazole. This suggests that clustering is actively maintained or enforced by the association of centromeres with microtubules throughout interphase. Unlike the Rabl-orientation known from many higher eukaryotes, centromere clustering in yeast is not only a relic of anaphase chromosome polarization, because it can be reconstituted without the passage of cells through anaphase. Within the rosette, homologous centromeres are not arranged in a particular order that would suggest somatic pairing or genome separation. PMID- 10806102 TI - Expression of autofluorescent proteins reveals a novel protein permeable pathway between cells in the lens core. AB - The lens of the eye is composed of concentric layers of tightly packed fiber cells. The oldest fibers, those in the lens core, lose their nuclei and other organelles during terminal differentiation. This is thought to ensure the clarity of the lens. The anucleated core fibers are sustained by gap junction-mediated communication with metabolically active cells near the lens surface. In this study, we expressed autofluorescent proteins and microinjected fluorescent markers to probe cell-to-cell communication in different regions of the developing lens. Our data indicate that a novel cell-cell diffusion pathway becomes patent in the lens core during development. This pathway is remarkable in that it is permeable to proteins and other large molecules and is thus distinct from gap junctions. Diffusion of large molecules probably occurs through regions of membrane fusion observed between neighboring cells in the lens core. Further direct evidence for a continuous plasma membrane system was provided by the observation that exogenous membrane proteins expressed in one core fiber cell were able to diffuse laterally into the membranes of adjacent fibers. Thus, the lens core appears to represent a true syncytium within which both membrane proteins and cytoplasmic proteins freely diffuse. Significantly, the outermost edge of the core syncytium encompasses a shell of nucleated, transcriptionally competent, fiber cells. This arrangement could facilitate the delivery of newly synthesized protein components to the aged and metabolically quiescent cells in the center of the lens. PMID- 10806103 TI - Yeast colonies synchronise their growth and development. AB - The ability to emit and receive signals over long distances is one of the characteristic attributes of multicellular organisms. Such communication can be mediated in different manners (by chemical compounds, light waves, acoustic waves etc.) and usually is reflected in the behaviour of the communicating organisms. Recently, we reported that individual yeast colonies, organised multicellular structures, can also communicate at long distance by means of volatile ammonia, which is produced by colonies in pulses separated by acidification of the medium. Here, we demonstrate that the colony that first reached the stage of intense ammonia production induces ammonia production response in surrounding colonies regardless of their age, causing the synchronisation of their NH(3) pulses and, consequently, the mutual affection of their growth. Also an artificial source of ammonia (but neither NH(4)(+) nor NaOH gradients) can immediately induce the ammonia production even in the colony starting its acidic stage of the development. The repeated transition of Candida mogii colonies from the acidic phase to the phase of intensive ammonia production is accompanied by dramatic changes in colony morphology and also in cell morphology and growth. Relatively smooth colonies in the acidic phase are formed by growing pseudohyphae. After ammonia induction, pseudohyphae decompose into non-dividing yeast-like cells, which rearrange themselves into ruffled spaghetti-like structures. The synchronisation of colony growth, that also exists between yeast colonies of different genera, could be important in establishing their optimal distribution in a natural habitat. PMID- 10806104 TI - Chromatin-bound Cdc6 persists in S and G2 phases in human cells, while soluble Cdc6 is destroyed in a cyclin A-cdk2 dependent process. AB - Cdc6 is essential for the initiation of DNA replication in all organisms in which it has been studied. In addition, recombinant Cdc6 can stimulate initiation in G(1) nuclei in vitro. We have analysed the behaviour of recombinant Cdc6 in mammalian cell extracts under in vitro replication conditions. We find that Cdc6 is imported into the nucleus in G(1 )phase, where it binds to chromatin and remains relatively stable. In S phase, exogenous Cdc6 is destroyed in a process that requires import into the nucleus and phosphorylation by a chromatin-bound protein kinase. Recombinant cyclin A-cdk2 can completely substitute for the nucleus in promoting destruction of soluble Xenopus and human Cdc6. Despite this regulated destruction, endogenous Cdc6 persists in the nucleus after initiation, although the amount falls. Cdc6 levels remain constant in G(2) then fall again before mitosis. We propose that cyclin A-cdk2 phosphorylation results in destruction of any Cdc6 not assembled into replication complexes, but that assembled proteins remain, in the phosphorylated state, in the nucleus. This process could contribute to the prevention of reinitiation in human cells by making free Cdc6 unavailable for re-assembly into replication complexes after G(1) phase. PMID- 10806105 TI - HZwint-1, a novel human kinetochore component that interacts with HZW10. AB - HZwint-1 (Human ZW10 interacting protein-1) was identified in a yeast two hybrid screen for proteins that interact with HZW10. HZwint-1 cDNA encodes a 43 kDa protein predicted to contain an extended coiled-coil domain. Immunofluorescence studies with sera raised against HZwint-1 protein revealed strong kinetochore staining in nocodazole-arrested chromosome spreads. This signal co-localizes at the kinetochore with HZW10, at a position slightly outside of the central part of the centromere as revealed by staining with a CREST serum. The kinetochore localization of HZwint-1 has been confirmed by following GFP fluorescence in HeLa cells transiently transfected with a plasmid encoding a GFP/HZwint-1 fusion protein. In cycling HeLa cells, HZwint-1 localizes to the kinetochore of prophase HeLa cells prior to HZW10 localization, and remains at the kinetochore until late in anaphase. This localization pattern, combined with the two-hybrid results, suggests that HZwint-1 may play a role in targeting HZW10 to the kinetochore at prometaphase. HZwint-1 was also found to localize to neocentromeres and to the active centromere of dicentric chromosomes. HZwint-1 thus appears to associate with all active centromeres, implying that it plays an important role in correct centromere function. PMID- 10806106 TI - Talin controls the exit of the integrin alpha 5 beta 1 from an early compartment of the secretory pathway. AB - Talin is a major cytosolic protein that links the intracellular domains of beta1 and beta3 integrins to the cytoskeleton. It is required for focal adhesion assembly. However, its downregulation not only slows down cell spreading and organization of focal adhesions but also impairs the maturation of some beta1 integrins, including the fibronectin receptor alpha5beta1. To investigate this, we characterized the beta1 integrin synthesized in cells expressing talin anti sense RNA (AT22 cells). We identified a large intracellular pool of beta1 integrins that is abnormally accumulated in an earlier compartment of the secretory pathway. In this report, we show that in talin-deficient AT22 cells, the aberrant glycosylation of integrin receptors is accompanied by a delay in the export of the integrin alpha5beta1. In normal cells, talin was found associated with beta1 integrins in an enriched membrane fraction containing Golgi and endoplasmic reticulum. Finally, microinjection of anti-talin antibodies resulted in accumulation of the integrins within the cells. These data strongly suggest that talin plays a specific role in the export of newly synthesized integrins. We propose that talin binding to the integrin may disclose a diphenylalanine export signal, which is present in the membrane-proximal GFFKR motif conserved in all integrin alpha chains. PMID- 10806107 TI - Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas. AB - A key event in deflagellation or deciliation is the severing of the nine outer doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+) dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing. PMID- 10806108 TI - The NIMA-related kinase X-Nek2B is required for efficient assembly of the zygotic centrosome in Xenopus laevis. AB - Nek2 is a mammalian cell cycle-regulated serine/threonine kinase that belongs to the family of proteins related to NIMA of Aspergillus nidulans. Functional studies in diverse species have implicated NIMA-related kinases in G(2)/M progression, chromatin condensation and centrosome regulation. To directly address the requirements for vertebrate Nek2 kinases in these cell cycle processes, we have turned to the biochemically-tractable system provided by Xenopus laevis egg extracts. Following isolation of a Xenopus homologue of Nek2, called X-Nek2B, we found that X-Nek2B abundance and activity remained constant through the first mitotic cycle implying a fundamental difference in Nek2 regulation between embryonic and somatic cell cycles. Removal of X-Nek2B from extracts did not disturb either entry into mitosis or the accompanying condensation of chromosomes providing no support for a requirement for Nek2 in these processes at least in embryonic cells. In contrast, X-Nek2B localized to centrosomes of adult Xenopus cells and was rapidly recruited to the basal body of Xenopus sperm following incubation in egg extracts. Recruitment led to phosphorylation of the X-Nek2B kinase. Most importantly, depletion of X-Nek2B from extracts significantly delayed both the assembly of microtubule asters and the recruitment of gamma-tubulin to the basal body. Hence, these studies demonstrate that X-Nek2B is required for efficient assembly of a functional zygotic centrosome and highlight the possibility of multiple roles for vertebrate Nek2 kinases in the centrosome cycle. PMID- 10806109 TI - The phosphatase inhibitor okadaic acid induces AQP2 translocation independently from AQP2 phosphorylation in renal collecting duct cells. AB - Phosphorylation by kinases and dephosphorylation by phosphatase markedly affect the biological activity of proteins involved in intracellular signaling. In this study we investigated the effect of the serine/threonine phosphatase inhibitor okadaic acid on water permeability properties and on aquaporin2 (AQP2) translocation in AQP2-transfected renal CD8 cells. In CD8 cells both forskolin alone and okadaic acid alone increased the osmotic water permeability coefficient P(f) by about 4- to 5-fold. In intact cells, in vivo phosphorylation studies revealed that forskolin stimulation resulted in a threefold increase in AQP2 phosphorylation. In contrast, okadaic acid treatment promoted only a 60% increase in AQP2 phosphorylation which was abolished when this treatment was performed in the presence of 1 microM H89, a specific protein kinase A (PKA) inhibitor. Nevertheless, in this latter condition, confocal microscopy analysis revealed that AQP2 translocated and fused to the apical membrane. Okadaic acid-induced AQP2 translocation was dose dependent having its maximal effect at a concentration of 1 microM. In conclusion, our results clearly indicate that okadaic acid exerts a full forskolin-like effect independent from AQP2 phosphorylation. Thus AQP2 phosphorylation is not essential for water channel translocation in renal cells, indicating that different pathways might exist leading to AQP2 apical insertion and increase in P(f). PMID- 10806110 TI - Disruption of Golgi structure and function in mammalian cells expressing a mutant dynamin. AB - The large GTPase dynamin is a mechanoenzyme that participates in the scission of nascent vesicles from the plasma membrane. Recently, dynamin has been demonstrated to associate with the Golgi apparatus in mammalian cells by morphological and biochemical methods. Additional studies using a well characterized, cell-free assay have supported these findings by demonstrating a requirement for dynamin function in the formation of clathrin-coated, and non clathrin-coated vesicles from the trans-Golgi network (TGN). In this study, we tested if dynamin participates in Golgi function in living cells through the expression of a dominant negative dynamin construct (K44A). Cells co-transfected to express this mutant dynamin and a GFP-tagged Golgi resident protein (TGN38) exhibit Golgi structures that are either compacted, vesiculated, or tubulated. Electron microscopy of these mutant cells revealed large numbers of Golgi stacks comprised of highly tubulated cisternae and an extraordinary number of coated vesicle buds. Cells expressing mutant dynamin and GFP-tagged VSVG demonstrated a marked retention (8- to 11-fold) of the nascent viral G-protein in the Golgi compared to control cells. These observations in living cells are consistent with previous morphological and in vitro studies demonstrating a role for dynamin in the formation of secretory vesicles from the TGN. PMID- 10806111 TI - Genetic defects in acetylcholine signalling promote protein degradation in muscle cells of Caenorhabditis elegans. AB - A myosin-lacZ fusion, expressed in 103 muscle cells of Caenorhabditis elegans, reports on how proteolysis in muscle is controlled by neural and intramuscular signals. Upon acute starvation, the fusion protein is degraded in the posterior 63 cells of the body-wall muscle, but remains stable in 32 anterior body-wall muscles and 8 vulval muscle cells. This distinction correlates with differences in the innervation of these cells. Reporter protein in the head and vulval muscles becomes labile upon genetic 'denervation' in mutants that have blocks in pre-synaptic synthesis or release of acetylcholine (ACh) or post-synaptic reception at nicotinic ACh receptors (nAChR), whereas protein in all 103 muscles is stabilized by the nicotinic agonist levamisole in the absence of ACh production. Levamisole does not stabilize muscle protein in nAChR mutants that are behaviorally resistant to levamisole. Neural inputs thus exert negative control over the proteolytic process in muscle by stimulating muscle nicotinic ACh receptors. PMID- 10806112 TI - The multidrug-resistant phenotype associated with overexpression of the new ABC half-transporter, MXR (ABCG2). AB - Mechanisms of drug resistance other than P-glycoprotein are of increasing interest as the list of newly identified members of the ABC transport family has grown. We sought to characterize the phenotype of the newly discovered ABC transporter encoded by the mitoxantrone resistance gene, MXR, also known as ABCP1 or BCRP. The pharmacodynamics of mitoxantrone and 12 other fluorescent drugs were evaluated by confocal microscopy in four multidrug-resistant human colon (S1) and breast (MCF-7) cancer cell lines. We utilized two sublines, MCF-7 AdVp3000 and S1 M1-80, and detected overexpression of MXR by PCR, immunoblot assay and immunohistochemistry. These MXR overexpressing sublines were compared to cell lines with P-glycoprotein- and MRP-mediated resistance. High levels of cross resistance were observed for mitoxantrone, the anthracyclines, bisantrene and topotecan. Reduced levels of mitoxantrone, daunorubicin, bisantrene, topotecan, rhodamine 123 and prazosin were observed in the two sublines with high MXR expression. Neither the P-glycoprotein substrates vinblastine, paclitaxel, verapamil and calcein-AM, nor the MRP substrate calcein, were extruded from MCF-7 AdVp3000 and S1-M1-80 cells. Thus, the multidrug-resistant phenotype due to MXR expression is overlapping with, but distinct from, that due to P-glycoprotein. Further, cells that overexpress the MXR protein seem to be more resistant to mitoxantrone and topotecan than cells with P-glycoprotein-mediated multidrug resistance. Our studies suggest that the ABC half-transporter, MXR, is a potent, new mechanism for conferring multiple drug resistance. Definition of its mechanism of transport and its role in clinical oncology is required. PMID- 10806113 TI - Identification of a novel C-terminal variant of beta II spectrin: two isoforms of beta II spectrin have distinct intracellular locations and activities. AB - It is established that variations in the structure and activities of betaI spectrin are mediated by differential mRNA splicing. The two betaI spectrin splice forms so far identified have either long or short C-terminal regions. Are analogous mechanisms likely to mediate regulation of betaII spectrins? Thus far, only a long form of betaII spectrin is reported in the literature. Five human expressed sequence tags indicated the existence of a short splice variant of betaII spectrin. The occurrence and DNA sequence of the short C-terminal variant was confirmed by analysis of human and rat cDNA. The novel variant lacks a pleckstrin homology domain, and has 28 C-terminal residues not present in the previously recognized longer form. Transcripts of the short C-terminal variant (7.5 and 7. 0 kb) were most abundant in tissues originating from muscle and nervous system. Antibodies raised to a unique sequence of short C-terminal variant recognized 240 kDa polypeptides in cardiac and skeletal muscle and in nervous tissue; in cerebellum and forebrain, additional 270 kDa polypeptides were detected. In rat heart and skeletal muscle, both long and short C-terminal forms of betaII spectrin localized in the region of the Z line. The central region of the sarcomere, coincident with the M line, was selectively labeled with antibodies to the short C-terminal form. In cerebellum, the short form was not detectable in parallel fibers, structures in which the long form was readily detected. In cultured cerebellar granule neurons, the long form was dominant in neurites, with the short form being most abundant in cell bodies. In vitro, the short form was found to lack the binding activity for the axonal protein fodaxin, which characterizes the C-terminal region of the long form. Subcellular fractionation of brain revealed that the short form was scarcely detectable in post-synaptic density preparations, in which the long form was readily detected. We conclude that variation in the structure of the C-terminal regions of betaII spectrin isoforms correlates with their differential intracellular targeting. PMID- 10806114 TI - The LIM and SH3 domain-containing protein, lasp-1, may link the cAMP signaling pathway with dynamic membrane restructuring activities in ion transporting epithelia. AB - Lasp-1 is a unique LIM and src homology 3 (SH3) domain-containing protein that was initially identified as a 40 kDa cAMP-dependent phosphoprotein in the HCl secreting gastric parietal cell. Because cAMP is a potent stimulator of parietal cell acid secretion, we have hypothesized that changes in lasp-1 phosphorylation might be involved in the regulation of ion transport-related activities, perhaps by modulating interactions among cytoskeletal and/or vesicle-associated proteins. In this study, we demonstrate that the cAMP-dependent acid secretory agonist, histamine, induces a rapid, sustained rise in parietal cell lasp-1 phosphorylation and this increase in phosphorylation is closely correlated with the acid secretory response. In addition, elevation of intracellular cAMP concentrations appear to induce a partial redistribution of lasp-1 from the cell cortex, where it predominates along with the gamma-isoform of actin in unstimulated cells, to the beta-actin enriched, apically-directed intracellular canalicular region, which is the site of active proton transport in the parietal cell. Additional studies demonstrate that although lasp-1 mRNA and protein are expressed in a wide range of tissues, the expression is specific for certain actin-rich cell types present within these tissues. For example, gastric chief cells, which contain relatively little F-actin and secrete the enzyme, pepsinogen, by regulated exocytosis, do not appear to express lasp-1. Similarly, lasp-1 was not detected in pancreatic acinar cells, which secrete enzymes by similar mechanisms and also contain relatively low levels of F-actin. Lasp-1 also was not detectable in proximal tubules in the kidney, in gastrointestinal smooth muscle, heart or skeletal muscle. In contrast, expression was prominent in the cortical regions of ion-transporting duct cells in the pancreas and in the salivary parotid gland as well as in certain F-actin-rich cells in the distal tubule/collecting duct. Interestingly, moderate levels of expression were also detected in podocytes present in renal glomeruli and in vascular endothelium. In primary cultures of gastric fibroblasts, lasp-1 was present mainly within the tips of lamellipodia and at the leading edges of membrane ruffles. Taken together these results support the hypothesis that the lasp-1 plays an important role in the regulation of dynamic actin-based, cytoskeletal activities. Agonist-dependent changes in lasp-1 phosphorylation may also serve to regulate actin-associated ion transport activities, not only in the parietal cell but also in certain other F actin-rich secretory epithelial cell types. PMID- 10806115 TI - An isoform of kinesin light chain specific for the Golgi complex. AB - Conventional kinesin is a motor protein implicated in the transport of a variety of cytoplasmic organelles along microtubules. The kinesin molecule consists of two heavy chains with motor domains at their amino termini and two light chains, which, together with the carboxyl termini of the heavy chains, are proposed to mediate binding to cargoes. Since the light chains are represented by multiple isoforms diverging at their carboxyl termini they are presumed to specify kinesin targeting to organelles. Previously, we isolated five cDNAs, encoding hamster kinesin light chain isoforms, and found that one of them (B or C) preferentially associated with mitochondria. To obtain additional evidence proving the specific location of various kinesin light chain isoforms on organelles, we made an antibody against a 56 amino-acid sequence found at the carboxyl-terminal regions of the hamster D and E isoforms. By indirect immunofluorescence, this antibody specifically labeled the Golgi complex in cultured cells. In western blots of total cell homogenates, it recognized two close polypeptides, one of which co purified with the Golgi membranes. Thus, the results of this and previous studies demonstrate that different kinesin light chains are associated with different organelles in cells. PMID- 10806116 TI - Collagen degradation and platelet-derived growth factor stimulate the migration of vascular smooth muscle cells. AB - Cell migration is a key event in many biological processes and depends on signals from both extracellular matrix and soluble motogenic factors. During atherosclerotic plaque development, vascular smooth muscle cells migrate from the tunica media to the intima through a basement membrane and interstitial collagenous matrix and proliferate to form a neointima. Matrix metalloproteinases have previously been implicated in neointimal formation and in this study smooth muscle cell adhesion and migration on degraded collagen have been evaluated. Vascular smooth muscle cells adhered to native intact collagen type I and to its first degradation by-product, 3/4 fragment (generated by collagenase-3 cleavage), unwound at 35 degrees C to mimic physiological conditions. PDGF-BB pre-treatment induced a fourfold stimulation of smooth muscle cell motility on the collagen 3/4 fragment whereas no increase in smooth muscle cell motility on collagen type I was observed. Cell migration on collagen type I was mediated by alpha2 integrin, whereas PDGF-BB-stimulated migration on the 3/4 collagen fragment was dependent on alphavbeta3 integrin. alphavbeta3 integrin was organised in clusters concentrated at the leading and trailing edges of the cells and was only expressed when cells were exposed to the 3/4 collagen fragment. Tyrphostin A9, an inhibitor of PDGF receptor-beta tyrosine kinase activity, resulted in complete abolition of migration of PDGF-BB treated cells on collagen type I and 3/4 fragment. These results strongly support the hypothesis that the cellular migratory response to soluble motogens can be regulated by proteolytic modification of the extracellular matrix. PMID- 10806117 TI - LdARL-3A, a Leishmania promastigote-specific ADP-ribosylation factor-like protein, is essential for flagellum integrity. AB - The small G protein-encoding LdARL-3A gene, a homologue of the human ARL-3 gene, was isolated from Leishmania donovani, and its protein product characterised. It is unique in the Leishmania genome and expressed only in the extracellular promastigote insect form, but not in the intracellular amastigote mammalian form, as shown by northern blots and western blots developed with a specific anti-C terminus immune serum. Indirect immunofluorescence microscopy revealed distinct labelled spots regularly distributed on the plasma membrane, including the part lining the flagellum and the flagellar pocket. By transfection experiments, it was found that wild-type LdARL-3A-overexpressing promastigotes reached higher densities in culture, but released significantly less secreted acid phosphatase in the extracellular medium than the parental strain. When LdARL-3A blocked under the GDP-bound 'inactive' form or with an inactivated potential myristoylation site was overexpressed, the cells displayed an apparent wild-type phenotype, but died earlier in the stationary phase; in contrast to parental cells, they showed a diffuse pattern of fluorescence labelling in the cytoplasm and on the cell membrane. Strikingly, when a constitutively 'active' form of LdARL-3A (blocked under the GTP-bound form) was overexpressed, the promastigotes were immobile with a very short flagellum, a slow growth rate and a low level of acid phosphatase secretion; the length of the flagellum was inversely proportional to mutant protein expression. We concluded that LdARL-3A could be an essential gene involved in flagellum biogenesis; it may provide new approaches for control of the parasite at the insect stage. PMID- 10806118 TI - Colocalization of chaperone Cpn60, proinsulin and convertase PC1 within immature secretory granules of insulin-secreting cells suggests a role for Cpn60 in insulin processing. AB - Many of the mechanisms that control insulin processing and packaging by interaction with different elements along the secretory pathway remain poorly understood. We have investigated the possibility that Cpn60, a member of the heat shock protein family, may be present in rat insulin-secreting cells, participating in the proinsulin-insulin maturation process. Immunofluorescence and high resolution immunocytochemical studies revealed the presence of the Cpn60 protein all along the insulin secretory pathway, being particularly abundant over the proinsulin-containing immature secretory granules. Double-labeling experiments showed associations between Cpn60 and proinsulin, as well as between Cpn60 and PC1 convertase, with a preferential binding to proinsulin. These findings paralleled those of coimmunoprecipitation studies showing the Cpn60 chaperone and the mature form of the PC1 convertase in proinsulin immunoprecipitates, as well as the PC1 in Cpn60 immunoprecipitates from total islet cell extracts. In vitro binding of Cpn60 to proinsulin, insulin and glucagon was also documented. Cpn60, significantly abundant in proinsulin containing secretory granules where conversion of proinsulin to insulin takes place, and the colocalization of the chaperone with proinsulin and PC1 convertase suggest that the Cpn60 protein may play a role directing precise molecular interactions during insulin processing and/or packaging. PMID- 10806119 TI - Expression from the human occludin promoter is affected by tumor necrosis factor alpha and interferon gamma. AB - The 65 kDa protein occludin is a membrane-spanning part of the epithelial tight junction, which is the main barrier of the paracellular pathway. The function of occludin as part of tight junctions is still poorly understood and even less is known about the regulatory mechanisms that influence occludin gene expression. This study aimed to identify the sequences essential in cis for genomic regulation of tight junction formation and to investigate their funcional role in cytokine-dependent tight junction regulation. Using genome walking cloning of occludin-specific human genomic DNA sequences, a 1853 bp DNA fragment containing the transcription start point of occludin cDNA sequences was amplified and sequenced. Subcloning of this fragment in front of the luciferase reporter gene revealed strong expression of enzymatic activity after transfection of the human intestinal cell line HT-29/B6. With subsequent deletions of parts of the promoter fragment, its size was reduced to 280 bp that are necessary and sufficient to mediate promoter activity. Tumor necrosis factor alpha and another cytokine involved in inflammation, interferon gamma, reduced transepithelial resistance in HT-29/B6 cells, which was preceded by a decrease in occludin mRNA expression as revealed by northern blot analysis. Tumor necrosis factor alpha and interferon gamma diminished occludin promoter activity alone and even synergistically, suggesting a genomic regulation of alterations of the paracellular barrier. In conclusion, proinflammatory cytokines such as tumor necrosis factor alpha and interferon gamma can downregulate the expression of the transmembrane tight junction strand protein occludin, paralleling the barrier disturbance detected electrophysiologically. This could be an important mechanism in gastrointestinal diseases accompanied by barrier defects, for example inflammatory bowel diseases. PMID- 10806120 TI - Renal involvement in HIV-infection. Results from the Frankfurt AIDS Cohort Study (FACS) and a review of the literature. AB - The current report describes the experience from the Frankfurt AIDS Cohort Study with patients suffering from renal failure. The clinical data of 4993 HIV infected patients between 1983 and 1998 were analyzed retrospectively. Patients were seen at least twice a year and clinical features, routine laboratory results, including CD4+ cell counts, concomittant diseases, and antiretroviral therapy were documented by standardized methods. The incidence of renal failure during 4 observation periods with different antiretroviral treatment strategies are compared and data are discussed. Within the 16 years of observation 47 patients with impairement of their kidney function were identified. A trend to an increase of RF could be documented (chi superset2 -for trend p = 0.0246). The additional review intends to summarize the diverse reasons leading to renal dysfunction in HIV-infected individuals with special emphasis on glomerular disease and renal complications related to HIV therapy. PMID- 10806121 TI - Indication, outcome and follow up of intensive care in patients with HIV infection. AB - BACKGROUND/AIM: The admission to intensive care is controversially discussed in patients with HIV infection, since life expectancy is limited. Therefore, we analyzed indications, outcomes and follow up of all patients with confirmed HIV infection and Aids defining symptoms who had been admitted to the intensive care unit (ICU) of our department between 1985-1996. RESULTS: 49 patients were admitted to the ICU, 42 of them with CDC stage C of HIV infection before admission. The leading indications were pneumonia (n = 15; PCP: 10, bacterial: 5), acute bleedings (n = 14), acute neurological diseases (n = 6), and gastrointestinal perforation (n = 5). Overall mortality was 39% (19/49) with a higher mortality seen in patients with respiratory disorders (53%) compared to non-respiratory disorders (22%, n.s.). The only significant predictor of mortality was the serum creatinine (p = 0.001), while differences in the APACHE II score between survivors and non-survivors did not reach statistical significance (22 +/- 7, 16 +/- 5; p = 0.14). During follow up no difference was seen in the life expectancy of HIV-infected survivors of intensive care as compared to those patients with AIDS who had never been admitted to ICU (8.4 months versus 9 months). DISCUSSION: The need for intensive care in HIV infected patients does not accelerate the progression of HIV infection to death, if the complications requiring ICU intervention can be managed successfully. Respiratory infections and impaired renal function are risk factors for a fatal outcome. Thus, HIV infected patients benefit from intensive care therapy. PMID- 10806122 TI - Treatment of candidal infections with fluconazole in neonates and infants. AB - To study the therapy, efficacy and safety of fluconazole in candidal mycoses during neonatal phase and infancy a case review in 53 newborns and infants was performed. The majority of these patients were premature with a median birth weight of 1120 g and born within gestational week 23-38. The median age at the onset of fluconazole treatment was 5 weeks. All patients had underlying diseases and several risk factors, which favored the occurence of a systemic candidal mycosis. Systemic candidiasis was the most frequent diagnosis (75.5%). Fluconazole was administered at a daily dosage of 5-6 mg/kg for a median duration of 21 days. The hepatic, renal and hematologic functions were assessed before, one, two, and three weeks after start of treatment. Yeasts were identified in 37 patients. The most common fungus isolated at baseline was Candida albicans (68%). Clinical cure or improvement was reported in 31 out of 38 patients (81.6%). Mycological cure was achieved in 25 out of 32 newborns and infants. Despite the limited number of patients with outcome data, these preliminary results of a small cohort clearly indicate the effective antifungal therapy with fluconazole in neonates and infants. No serious side effects were observed in fluconazole treated patients. Two patients with megaureter-megacystis-hydronephrosis syndrome and severe meningoencephalitis showed a mild increase in liver enzymes. - CONCLUSION: Fluconazole seems to be an effective therapy for systemic and other forms of candidiasis in infants including very low birth weight infants (VLBWI; <1500 g). These favorable safety and efficacy data are similar to results obtained with fluconazole in older children and adults. These findings, however, must be supported by larger trials. The recommended daily dose is 5 mg/kg body weight. Only in VLBWI the dosage interval within the first two weeks of life should be prolonged up to 3 days and fluconazole serum levels should be monitored. PMID- 10806123 TI - Perfluorocarbons useful tools for medicine. AB - Perfluorocarbons (PFCs) combine rather unique chemical and physical properties together with physiological inertness. Due to this, they have become useful tools in medicine. Whereas the majority of applications benefit from their excellent oxygen solubility, there are several applications making use of other PFC properties. The great importance of PFC ultra-purity is especially emphasized. PMID- 10806124 TI - Influence of smoking on cotinine and caffeine plasma levels in patients with alcoholic liver cirrhosis. AB - BACKGROUND: Retention of caffeine was observed in patients with alcoholic liver cirrhosis and impaired liver function. Cotinine, the major metabolite of nicotine, is transformed by microsomal N-oxidation to secondary metabolites. The aim of this study was to investigate if impaired liver function leads to a retention of cotinine in a similar way to caffeine retention. Furthermure the influence of smoking on cotinine and caffeine plasma levels was studied. METHODS: 91 smokers and 12 nonsmokers with alcoholic liver cirrhosis were subdivided according to their smoking habits. Cotinine plasma levels and fasting caffeine concentrations were measured by a gaschromatographic method. Concentrations of conjugated bile acids were measured by RIA. 10 healthy smokers and 11 nonsmokers were used as a control group. RESULTS: Mean plasma cotinine concentrations found in slight smokers (200 +/- 155 ng/ml), intermediate smokers (384 +/- 223 ng/ml) and heavy smokers (430 +/- 266 ng/ml) with alcoholic liver cirrhosis were significantly higher than in healthy, smoking volunteers with slight, intermediate, and heavy smoking (101 +/- 14; 274 +/- 112; 345 +/- 85 ng/ml) (p <0.01) respectively. In nonsmokers with alcoholic liver cirrhosis plasma cotinine (44 +/- 25 ng/ml) was significantly elevated compared to healthy nonsmokers (27 +/- 19 ng/ml) (p <0.01). - Fasting caffeine plasma levels in patients with alcoholic liver cirrhosis (4.00 +/- 5. 20 microg/ml) were significantly higher than in healthy volunteers (0.91 +/- 0.42 microg/ml) (p <0.01). A decrease of plasma levels was observed in correlation to the amount of smoking in patients with alcoholic cirrhosis (slight smokers: 7.67 +/- 8.54 microg/ml, intermediate smokers: 3.35 +/- 2.91 microg/ml and heavy smokers: 2. 48 +/- 2.68 microg/ml). Conjugated bile acids were elevated in patients with alcoholic liver cirrhosis to 32,56 +/- 38,24 mmol/l. CONCLUSIONS: Increased cotinine plasma levels in smokers and nonsmokers with alcoholic liver cirrhosis demonstrate a cotinine retention in patients with impaired liver function. The inducing effect of smoking is shown by a decrease of fasting caffeine plasma concentrations. PMID- 10806125 TI - Reduced myocardial cellular damage and lipid peroxidation in off-pump versus conventional coronary artery bypass grafting. AB - BACKGROUND: The development of new surgical devices and techniques allows off pump coronary artery bypass grafting (OPCAB) without the use of CPB and cardioplegia. This study tested whether OPCAB reduces myocardial cell damage, lipid peroxidation and systemic endothelin release when compared to conventional coronary artery bypass grafting. METHODS: Twenty-six patients were assigned to either the OPCAB procedure using a suction device and regular sternotomy (n = 13) or were treated conventionally using extracorporeal circulation, blood cardioplegia and hypothermic arrest (29-31 degrees C; n = 13). Troponin I and creatine kinase - MB were determined for cardiac specific cell damage. Myocardial and systemic malondialdhyde levels were measured to account for oxyradical mediated lipid peroxidation. Systemic big-endothelin levels were determined as a marker for endothelial cell activation. RESULTS: A significant reduction of the cardiac specific cell damage was observed in the OPCAB group vs. the CABG group over time in the absence of acute myocardial ischemia or infarction. In addition, systemic and myocardial lipid peroxidation as measured by the malondialdehyde (MDA) levels were lower in the OPCAB group when compared to CABG. Finally, plasma levels of big-Endothelin (big-ET) significantly rose in the CABG but not in the OPCAB group. CONCLUSIONS: The data of the present study indicate that OPCAB revascularization without the use of CPB and cardioplegic arrest reduces myocardial cell damage and lipid peroxidation. It is also associated with a reduced activation of the potent vasoconstrictor peptide endothelin. All of this may contribute to improved myocardial function and faster postoperative recovery from surgical revascularization procedures, particularly in critically ill patients. PMID- 10806126 TI - Pancreatitis following hepatitis A vaccination. AB - We describe a 23-year-old male patient who presented with epigastric abdominal pain, 8 days following vaccination with inactivated hepatitis A virus (Haverix(R)). Clinical and laboratory data confirmed the diagnosis of pancreatitis. Repeat polymerase chain reaction (PCR) for hepatitis A replication was negative. A comprehensive evaluation ruled out other etiologies for pancreatitis. IgM Hepatitis A antibodies did not develop even after 3 months. Pancreatitis following Hepatitis A is a well-known complication of the viremia, but the exact mechanism is controversial. We suggest that the pancreatitis may have been a cellular immunlogical reaction to one of the antigens of hepatitis A virus vaccine, or it might have been caused by the release of mediators of anaphylaxis such as histamine and leucotriens, induced by HAV antigens, resulting in pancreatitis without development of humoral immunization. PMID- 10806127 TI - Sleep-disordered breathing and hypertension: more research is still needed. PMID- 10806128 TI - Upper airway resistance syndrome is a distinct syndrome. PMID- 10806129 TI - Upper airway resistance syndrome is not a distinct syndrome. PMID- 10806130 TI - Rebuttal from drs. guilleminault and chowdhuri PMID- 10806131 TI - Rebuttal from dr. douglas PMID- 10806132 TI - Recent advances in critical care gastroenterology. PMID- 10806133 TI - How science happened to me. PMID- 10806134 TI - Comparison of atenolol, amlodipine, enalapril, hydrochlorothiazide, and losartan for antihypertensive treatment in patients with obstructive sleep apnea. AB - We compared the effects of atenolol (50 mg), amlodipine (5 mg), enalapril (20 mg), hydrochlorothiazide (25 mg), and losartan (50 mg) given in once-daily oral doses on office and ambulatory blood pressures (BPs) in patients with hypertension and obstructive sleep apnea (OSA). Each of 40 randomized patients was treated in sequence with two of the five agents (balanced incomplete block design). Treatment periods lasted 6 wk and were separated by a 3-wk washout period. Changes in BP from baseline with the study substances were compared through analysis of variance. Office diastolic BP, our primary outcome variable, was most effectively lowered by atenolol, with all four post hoc differences between atenolol and the remaining substances being statistically significant. Reductions in office systolic and daytime ambulatory BP were not significantly different among the five compounds. However, atenolol reduced mean nighttime ambulatory diastolic and systolic BP more effectively than did amlodipine, enalapril, or losartan (but not hydrochlorothiazide). Severity of sleep disordered breathing and well-being during the day were not significantly influenced by any of the study compounds. Our findings are in accordance with the hypothesis that an overactivity of the sympathetic nervous system is an important mechanism behind the development or maintenance of hypertension in patients with OSA. PMID- 10806135 TI - The effect of intensive plasma water exchange by hemofiltration on hemodynamics and soluble mediators in canine endotoxemia. AB - High volume hemofiltration (HVHF) (200 ml/kg/h) improves hemodynamics in experimental septic shock but is difficult to apply clinically. Accordingly, we studied whether less intensive HVHF (80 ml/kg/h) can still improve hemodynamics in experimental septic shock. We also investigated its effect on the serum concentrations of several inflammatory mediators, including endothelin (ET-1), endotoxin (LPS), tumor necrosis factor-alpha (TNF-alpha), and 6-keto prostaglandin F(1alpha) (6-kepto PGF(1alpha)). Sixteen anesthetized dogs were connected to a continuous veno-venous hemofiltration (CVVH) (filtration: 80 ml/kg/h) or sham circuit and endotoxin (0.5 mg/kg) was infused intravenously over 5 min. Hemodynamic variables were measured at baseline and at 15, 45, 90, and 180 min. The major hemodynamic finding was that endotoxin-induced hypotension was significantly attenuated by intensive CVVH (p < 0.04). Changes in cardiac output and right ventricular ejection fraction were equal in both groups. ET-1 levels, but not LPS, TNF-alpha, or 6-keto PGF(1alpha), were lower during CVVH (p = 0.042). Endotoxin or TNF-alpha were not found in the ultrafiltrate. Median clearances of ET-1 and 6-keto PGF(1alpha) during intensive CVVH were 8.8 and 25.9 ml/m, respectively. We conclude that intensive CVVH attenuates the early component of endotoxin-induced hypotension and reduces serum concentrations of endothelin-1. The effect of CVVH on blood pressure is not explained by convective clearance of the mediators in question. PMID- 10806136 TI - Polymorphism at the glutathione S-transferase GSTP1 locus. A new marker for bronchial hyperresponsiveness and asthma. AB - Most genetic studies of asthma have concentrated on genes on chromosomes 11q and 5q and their association with the key asthma-related phenotypes of bronchial hyperresponsiveness (BHR) and atopy. Although asthma is characterized by airway inflammation, a critical component of which is oxidative stress, few data exist on genes involved in protecting against this insult. We describe an association study designed to examine whether allelic variation at the glutathione-S transferase GSTP1 locus influences expression of the BHR and atopy phenotypes in asthma. The enzyme encoded by GSTP1 utilizes a variety of lipid and DNA products of oxidative stress, and polymorphic variants of this gene are associated with altered catalytic function of this enzyme. We found that the frequency of GSTP1 Val(105)/Val(105) was significantly lower in asthmatic than in control subjects. Indeed, the presence of this genotype conferred a sixfold lower risk of asthma than did GSTP1 Ile(105)/Ile(105). Remarkably, asthma risk in Val(105) homozygotes was further reduced (by ninefold) after correction for atopic indices, age, and gender. Trend analysis after stratification according to the degree of bronchial reactivity/obstruction showed that the frequency of GSTP1 Val(105)/Val(105) correlates with decreasing severity of airway dysfunction. Furthermore, subjects with GSTP1 Val(105)/Val(105) have four- and 10-fold lower risks, respectively, of exhibiting atopy defined by skin test positivity and IgE level. These data show that GSTP1 polymorphism is strongly associated with asthma and related phenotypes, and provide an alternative explanation for the linkage of chromosome 11q13 with BHR and atopy. PMID- 10806137 TI - The incidence and pathogenesis of cardiopulmonary deterioration after abrupt withdrawal of inhaled nitric oxide. AB - We studied the effect of abrupt discontinuation of inhaled nitric oxide (iNO) in patients receiving this drug for treatment of acute hypoxemic respiratory failure (AHRF), in order to determine the need for continued therapy, the incidence and nature of adverse events, and the risk factors predicting these adverse events. Thirty-one patients who showed an initial increase in Pa(O(2)) of > 20 mm Hg in response to iNO underwent a discontinuation trial at 10 to 30 h after beginning iNO. Indwelling arterial and pulmonary artery catheters facilitated monitoring of hemodynamic and gas-exchange parameters. For the group, discontinuation of iNO caused a significant decrease in Pa(O2 ), arterial and mixed venous oxygen saturation, and ratio of Pa(O(2)) to fraction of inspired oxygen (FI(O(2))). Three patterns of response were observed. Eight of 31 (25.8%) patients had minimal changes in oxygenation or hemodynamics, suggesting no need for ongoing therapy. Fifteen of 31 (48%) patients had worsened gas exchange as a predominant response. Eight of 31 patients exhibited hemodynamic collapse, defined as > 20% fall in cardiac output and/or mean arterial blood pressure. In this last subgroup, the pattern of cardiovascular changes suggested that this response arose from an acute increase in right ventricular afterload, and was not a consequence of gas-exchange abnormalities. In all cases, reinstitution of iNO promptly reversed worsened hemodynamics and gas exchange. Independent factors associated with an increased risk of cardiovascular collapse included multisystem organ failure, older age, and initial blood pressure increase in response to iNO; a smaller change in the ratio of Pa(O(2)) to FI(O(2)) with initiation of iNO therapy also tended to correlate with this phenomenon. We conclude that careful and monitored discontinuation of iNO in patients with AHRF will identify substantial fractions of patients who are either receiving no benefit from this therapy or who require iNO to maintain an adequate circulation and are therefore at risk for adverse outcome with transport or inadvertent discontinuation of iNO. Future trials of iNO should recognize this complication of such therapy and include assessments for it. PMID- 10806138 TI - How is mechanical ventilation employed in the intensive care unit? An international utilization review. AB - A 1-d point-prevalence study was performed with the aim of describing the characteristics of conventional mechanical ventilation in intensive care units ICUs from North America, South America, Spain, and Portugal. The study involved 412 medical-surgical ICUs and 1,638 patients receiving mechanical ventilation at the moment of the study. The main outcome measures were characterization of the indications for initiation of mechanical ventilation, the artificial airways used to deliver mechanical ventilation, the ventilator modes and settings, and the methods of weaning. The median age of the study patients was 61 yr, and the median duration of mechanical ventilation at the time of the study was 7 d. Common indications for the initiation of mechanical ventilation included acute respiratory failure (66%), acute exacerbation of chronic obstructive pulmonary disease (13%), coma (10%), and neuromuscular disorders (10%). Mechanical ventilation was delivered via an endotracheal tube in 75% of patients, a tracheostomy in 24%, and a facial mask in 1%. Ventilator modes consisted of assist/control ventilation in 47% of patients and 46% were ventilated with synchronized intermittent mandatory ventilation, pressure support, or the combination of both. The median tidal volume setting was 9 ml/kg in patients receiving assist/control and the median setting of pressure support was 18 cm H(2)O. Positive end-expiratory pressure was not employed in 31% of patients. Method of weaning varied considerably from country to country, and even within a country several methods were in use. We conclude that the primary indications for mechanical ventilation and the ventilator settings were remarkably similar across countries, but the selection of modes of mechanical ventilation and methods of weaning varied considerably from country to country. PMID- 10806139 TI - Effects of terbutaline and budesonide on sputum cells and bronchial hyperresponsiveness in asthma. AB - Previous studies have shown that the regular administration of short acting beta agonists can be associated with adverse effects on airway caliber and bronchial hyperresponsiveness (BHR) and that this may occur through a proinflammatory mechanism. The aim was to explore possible adverse effects of high-dose beta agonist therapy and to assess any adverse interaction with corticosteroids. We undertook a randomized, crossover study to investigate the effects of 6 wk of treatment with regular terbutaline (1 mg four times a day), regular budesonide (400 microg twice a day), combined treatment, and placebo in subjects with mild to moderate asthma. Major endpoints were PD(15) saline, PD(20) methacholine, and induced sputum differential cell counts. Thirty-four subjects were randomized and 28 completed the study. PD(15) saline decreased on terbutaline alone compared with placebo treatment and on combined treatment compared with budesonide alone (mean fold decrease of 0.57 [95% CI = 0.36, 0.90] and 0.65 [95% CI = 0.43, 0.97], respectively). PD(20) methacholine was not affected by the use of terbutaline either alone or in combination with budesonide. The percentage of eosinophils in induced sputum increased during terbutaline treatment alone compared with placebo (median 8.3% versus 4.4%, p = 0.049). The addition of terbutaline to budesonide did not affect the percentage of eosinophils compared with budesonide treatment alone. These findings support the hypothesis that short-acting beta-agonists have a permissive effect on airway inflammation and that when used in high dose there may be an unfavorable interaction with inhaled corticosteroids. PMID- 10806140 TI - Gender differences in the polysomnographic features of obstructive sleep apnea. AB - We examined the influence of gender on the polysomnographic features of obstructive sleep apnea (OSA) in a retrospective study of 830 patients with OSA diagnosed by overnight polysomnography (PSG). The severity of OSA was determined from the apnea- hypopnea index (AHI) for total sleep time (AHI(TST)), and was classified as mild (5 to 25 events/h), moderate (26 to 50 events/h), and severe (> 50/events/h). Differences in OSA during different stages of sleep were assessed by comparing the AHI during non-rapid eye movement (NREM) (AHI(NREM)) and rapid eye movement (REM) (AHI(REM)) sleep and calculating the "REM difference" (AHI(REM) - AHI(NREM)). Additionally, each overnight polysomnographic study was classified as showing one of three mutually exclusive types of OSA: (1) mild OSA, which occurred predominantly during REM sleep (REM OSA); (2) OSA of any severity, which occurred predominantly in the supine position (S OSA); or (3) OSA without a predominance in a single sleep stage or body position (A OSA). The mean AHI(TST) for men was significantly higher than that for women (31.8 +/- 1.0 versus 20.2 +/- 1.5 events/h, p < 0. 001). The male-to-female ratio was 3.2:1 for all OSA patients, and increased from 2.2:1 for patients with mild OSA to 7.9:1 for those with severe OSA. Women had a lower AHI(NREM) than did men (14.6 +/- 1.6 versus 29.6 +/- 1.1 events/h, p < 0.001), but had a similar AHI(REM) (42.7 +/- 1.6 versus 39.9 +/- 1.2 events/h). Women had a significantly higher REM difference than did men (28.1 +/- 1.5 versus 10.3 +/- 1.1 events/h, p < 0.01). REM OSA occurred in 62% of women and 24% of men with OSA. S OSA occurred almost exclusively in men. We conclude that: (1) OSA is less severe in women because of milder OSA during NREM sleep; (2) women have a greater clustering of respiratory events during REM sleep than do men; (3) REM OSA is disproportionately more common in women than in men; and (4) S OSA is disproportionately more common in men than in women. These findings may reflect differences between the sexes in upper airway function during sleep in patients with OSA. PMID- 10806141 TI - Increased numbers of both airway basophils and mast cells in sputum after allergen inhalation challenge of atopic asthmatics. AB - Mast cells and basophils are metachromatic cells that participate in allergic inflammation. Allergen challenge to the airways of atopic asthmatic individuals increases levels of metachromatic cells, which may reflect an increase in mast cells, basophils, or both. We conducted a study to characterize the kinetics of basophil and mast cell recruitment to the airways of atopic asthmatic subjects after allergen inhalation challenge, using monoclonal antibodies specific for each type of cell. Of 19 subjects, 14 developed both early- and late-phase asthmatic responses (dual responders [DRs]), whereas five developed only early asthmatic responses (early responders [ERs]) after allergen inhalation. There was a significant increase in the number of sputum eosinophils (p < 0.002) and basophils (p < 0.002) at 7 h and 24 h after challenge in both ERs and DRs. There was also a significant increase in the number of activated eosinophils (p = 0. 00002) and mast cells (p = 0.009) in sputum at 7 h and 24 h after challenge in DRs, but not in ERs (p > 0.4). DRs had a significantly higher number of allergen induced sputum basophils than did ERs (p < 0.01), and sputum basophils correlated significantly with airway hyperresponsiveness (AHR) to methacholine at 24 h after challenge (r = 0.66, p = 0.002). DRs tended to have higher allergen-induced basophil levels than did ERs, which may contribute to the observed AHR. PMID- 10806142 TI - Airway hyperresponsiveness in elite athletes. AB - It has been suggested that high-level training could contribute to the development of airway hyperresponsiveness (AHR), but the comparative effects of different sports on airway function remains to be determined. We evaluated 150 nonsmoking volunteers 18 to 55 yr of age; 100 athletes divided into four subgroups of 25 subjects each according to the predominant estimated hydrocaloric characteristic of ambient air inhaled during training: dry air (DA), cold air (CA), humid air (HA) and a mixture of dry and humid air (MA), and 50 sedentary subjects. Each subject had a respiratory questionnaire, a methacholine challenge, allergy skin-prick tests, and heart rate variability recording for evaluation of parasympathetic tone. The athletes had a 49% prevalence of AHR (PC(20) < 16 mg/ml), with a mean PC(20) of 16.9 mg/ml, compared with 28% (PC(20): 35.4) in sedentary subjects (p = 0.009). The prevalence (%) of AHR and mean PC(20) (mg/ml) varied as followed in the four subgroups of athletes: DA: 32% and 30.9; CA: 52% and 15.8; HA: 76% and 7.3; and MA: 32% and 21.5 (p = 0.002). The estimated parasympathetic tone was higher in athletes (p < 0.001), but this parameter showed only a weak correlation with PC(20) (r = -0.17, p = 0.04). This study has shown a significantly higher prevalence of AHR in athletes than in the control group because of the higher prevalence in the CA and HA groups. Parasympathetic activity may act as modulator of airway responsiveness, but the increased prevalence of AHR in our athlete population may be related to the type and possibly the content of inhaled air during training. PMID- 10806143 TI - Recruitment maneuvers in three experimental models of acute lung injury. Effect on lung volume and gas exchange. AB - Recruitment maneuvers (RM), consisting of sustained inflations at high airway pressures, have been advocated as an adjunct to mechanical ventilation in acute respiratory distress syndrome (ARDS). We studied the effect of baseline ventilatory strategy and RM on end-expiratory lung volume (EELV) and oxygenation in 18 dogs, using three models of acute lung injury (ALI; n = 6 in each group): saline lavage (LAV), oleic acid injury (OAI), and intratracheal instillation of Escherichia coli (pneumonia; PNM). All three models exhibited similar degrees of lung injury. The PNM model was less responsive to positive end-expiratory pressure (PEEP) than was the LAV or OAI model. Only the LAV model showed an oxygenation response to increasing tidal volume (VT). After RM, there were transient increases in Pa(O(2)) and EELV when ventilating with PEEP = 10 cm H(2)O. At PEEP = 20 cm H(2)O the lungs were probably fully recruited, since the plateau airway pressures were relatively high ( approximately 45 cm H(2)O) and the oxygenation was similar to preinjury values, thus making the system unresponsive to RM. Sustained improvement in oxygenation after RM was seen in the LAV model when ventilating with PEEP = 10 cm H(2)O and VT = 15 ml/kg. Changes in EELV correlated with changes in Pa(O(2)) only in the OAI model with PEEP = 10 cm H(2)O. We conclude that responses to PEEP, VT, and RM differ among these models of ALI. RM may have a role in some patients with ARDS who are ventilated with low PEEP and low VT. PMID- 10806144 TI - Additive effects of phosphodiesterase-4 inhibition on effects of rSP-C surfactant. AB - We have tested the effects of combined treatment with a recombinant surfactant protein C based surfactant (rSP-C surfactant) containing a phosphodiesterase-4 (PDE-4) inhibitor, roflumilast, in a lung lavage model of acute lung injury. The following groups were tested: (1) controls receiving sham exposure; (2) PDE-4 inhibitor (6.0 mg/kg body weight, intratracheally) alone; (3, 4) rSP-C surfactant (25 and 100 mg phospholipids [PL] per kg body weight) alone; and (5, 6 ) treatment with rSP-C surfactant (25 and 100 mg PL per kg body weight) combined with the PDE-4 inhibitor at a dose of 6.0 mg/kg body weight. The different groups were compared with respect to improving oxygenation and histopathologic changes, e.g., hyaline membrane (HM) formation. Both doses of rSP-C surfactant improved oxygenation while even this high dose of the PDE-4 inhibitor alone did not influence oxygenation compared with untreated control animals. Addition of the PDE-4 inhibitor led to improved oxygenation based on both doses of rSP-C surfactant. The PDE-4 inhibitor alone prevented further HM formation and infiltration of neutrophil leukocytes. The rSP-C surfactant was able to prevent further HM formation. Based on both doses of rSP-C surfactant, addition of the PDE-4 inhibitor showed additional effects on oxygenation and inhibition of HM formation. The effects of combined treatment were superior to each treatment alone, leading to the conclusion that a rSP-C surfactant containing a PDE-4 inhibitor may act synergistically in this animal model of acute lung injury. We conclude that combined treatment with rSP-C surfactant and a PDE-4 inhibitor may be an effective treatment for patients with acute lung injury. PMID- 10806145 TI - Respiratory syncytial virus bronchiolitis in infancy is an important risk factor for asthma and allergy at age 7. AB - We previously reported an increased risk for bronchial obstructive disease and allergic sensitization up to age 3 in 47 children hospitalized with a respiratory syncytial virus (RSV) bronchiolitis in infancy compared with 93 matched control subjects recruited during infancy. The aims of the present study were to evaluate the occurrences of bronchial obstructive disease and allergic sensitization in these children at age 7(1)/ (2). All 140 children reported for the follow-up, which included physical examination, skin prick tests, and serum IgE tests for common food and inhaled allergens. The cumulative prevalence of asthma was 30% in the RSV group and 3% in the control group (p < 0.001), and the cumulative prevalence of "any wheezing" was 68% and 34%, respectively (p < 0.001). Asthma during the year prior to follow-up was seen in 23% of the RSV children and 2% in the control subjects (p < 0.001). Allergic sensitization was found in 41% of the RSV children and 22% of the control subjects (p = 0.039). Multivariate evaluation of possible risk factors for asthma and sensitization using a stepwise logistic statistical procedure for all 140 children showed that RSV bronchiolitis had the highest independent risk ratio for asthma (OR: 12.7, 95% CI 3.4 to 47.1) and a significantly elevated independent risk ratio for allergic sensitization (OR: 2.4, 95% CI 1.1 to 5.5). In conclusion, RSV bronchiolitis in infancy severe enough to cause hospitalization was highly associatied with the development of asthma and allergic sensitization up to age 7(1)/ (2). The results support the theory that the RSV influences the mechanisms involved in the development of asthma and allergy in children. PMID- 10806146 TI - Individual characteristics and quitting in apprentices exposed to high-molecular weight agents. AB - To determine whether specific immunological sensitization (SIS) and symptoms of allergy are associated with quitting apprenticeships with exposure to high molecular-weight (HMW) agents. In a cohort study of 769 apprentices starting career programs in animal health technology (AHT), pastry-making (PM), and dental hygiene technology (DHT), health status was assessed at baseline and yearly for up to 44 mo. Subjects who quit the apprenticeship were contacted and offered a last series of measurements as if they pursued. The effect of various factors-at baseline and during the career program-on quitting was examined. Eighty-nine of 769 subjects (11.6%) quit, of whom 74 participated in this study. In multivariate analysis, a history of hay fever (OR = 1.66, 95% CI = 1.0 to 2.75) and attending the pastry-making program (OR = 2.33, 95% CI = 1.11 to 4.91) were significant determinants for quitting. In the subgroup of AHT, SIS to laboratory animals, hay fever, and shortness of breath were significant determinants by univariate analysis; in PM, the only determinant was age; and in DHT, hay fever. Work related symptoms during training were not associated with quitting. In a multivariate logistic regression analysis in AHT, including characteristics at baseline and at follow-up, baseline SIS (OR = 2.8, 95% CI = 0.94 to 8.38) and symptoms of asthma in general during follow-up (OR = 2.2, 95% CI = 0.94 to 5.38) were the main determinants for quitting, though they were of borderline significance. We conclude that health-related self-selection out of an apprenticeship with exposure to HMW agents is likely to occur and to cause an attrition bias. PMID- 10806147 TI - Are elderly patients with obstructive airway disease being prematurely discharged? AB - Despite the temporal trend toward decreasing length of hospital stay for all medical conditions in North America, the effect of different lengths of hospitalization on short-term outcomes such as readmission or mortality has not been well studied. However, there is growing concern that very short stays in hospital may result in premature discharges, which may lead to worse outcomes for patients. We conducted a population-based study of elderly patients with obstructive airway disease in Ontario, Canada to test the hypothesis that very short initial hospital stays increase the short-term risk for readmission and mortality. Using a cohort of 32,384 elderly patients 65 yr of age or older, we compared 15-d rates of readmission and mortality among patients with different lengths of stay. Although patients with hospital stays of less than 4 d were younger and had fewer comorbidities, they were 39% (95% confidence interval [CI], 20% to 61%) more likely to be readmitted and 45% (95% CI, 9% to 92%) more likely to die within 15 d postdischarge compared with those who stayed 4 to 6 d. The risk was highest among patients whose stay was less than or equal to 1 hospital day; they had a 69% (95% CI, 32% to 117%) excess risk of readmission and a 2.08 (95% CI, 1.23 to 3.45) -fold increase in mortality compared with those who stayed in hospital for 2 d. This suggests that some elderly patients with obstructive airway disease may be being prematurely discharged. PMID- 10806148 TI - Monocyte IL-10 production during respiratory syncytial virus bronchiolitis is associated with recurrent wheezing in a one-year follow-up study. AB - Respiratory syncytial virus (RSV) bronchiolitis is associated with subsequent recurrent wheezing episodes. To determine whether cytokine responses during infection can be of predictive value for the development of recurrent wheezing, we performed a follow-up study in 50 hospitalized children with RSV bronchiolitis. Monocyte and lymphocyte cytokine responses in vitro were studied during the acute phase of disease, and again during the convalescent phase, 3 to 4 wk later. Monocyte cytokine responses, including interleukin-10 (IL-10), were measured in whole blood cultures, stimulated with lipopolysaccharide and interferon-gamma (LPS + IFN-gamma). In addition, T-cell cytokine responses, including IFN-gamma and IL-4 production, were measured in whole-blood cultures stimulated with phytohemagglutinin (PHA) or alphaCD2 + alphaCD28. Cytokine responses were analyzed in relation to the development of recurrent episodes of wheezing, documented by parents in a diary during a 1-yr follow-up period. IL-10 responses during the acute phase of RSV bronchiolitis were comparable to those in healthy control subjects. During the convalescent phase, IL-10 responses were significantly increased in patients as compared with those in healthy control subjects (p < 0.001). At follow-up, 27 children (58%) had recurrent episodes of wheezing. IL-10 levels, measured during the convalescent phase, were significantly higher in patients who developed recurrent wheezing during the year after RSV bronchiolitis than in patients without recurrent episodes of wheezing (p = 0.006). Moreover, IL-10 responses during the convalescent phase correlated significantly with the number of wheezing episodes (r = 0.42, n = 46, p = 0.004). Interestingly, no association was found between IFN-gamma responses, IL-4 responses, or IFNgamma/IL-4 ratios and recurrent wheezing. We conclude that monocyte IL-10 responses in vitro upon stimulation with nonspecific stimuli may have predictive value for the development of recurrent wheezing after RSV bronchiolitis. Moreover, our results indicate that not only allergen-driven Th2 cytokine responses can lead to asthmatic symptoms but also virus-induced changes in cytokine responses may result in asthmatic symptoms. PMID- 10806149 TI - The role of hypoventilation and ventilation-perfusion redistribution in oxygen induced hypercapnia during acute exacerbations of chronic obstructive pulmonary disease. AB - The detailed mechanisms of oxygen-induced hypercapnia were examined in 22 patients during an acute exacerbation of chronic obstructive pulmonary disease. Ventilation, cardiac output, and the distribution of ventilation-perfusion (V A/Q ) ratios were measured using the multiple inert gas elimination technique breathing air and then 100% oxygen through a nose mask. Twelve patients were classified as retainers (R) when Pa(CO(2)) rose by more than 3 mm Hg (8.3 +/- 5.6; mean +/- SD) after breathing 100% oxygen for at least 20 min. The other 10 patients showed a change in Pa(CO(2)) of -1.3 +/- 2.2 mm Hg breathing oxygen and were classified as nonretainers (NR). Ventilation fell significantly from 9.0 +/- 1.5 to 7.2 +/- 1.2 L/min in the R group breathing oxygen (p = 0.007), whereas there was no change in ventilation in the NR group (9.8 +/- 1.8 to 9.9 +/- 1.8 L/min). The dispersion of V A/Q ratios as measured by log SD of blood flow (log SD Q) increased significantly in both R (0.96 +/- 0. 17 to 1.13 +/- 0.17) and NR (0.77 +/- 0.20 to 1.04 +/- 0.23, p < 0.05) groups breathing oxygen, whereas log SD of ventilation (log SD Q ) increased only in the R group (0.97 +/- 0.24 to 1.20 +/- 0.46, p < 0.05). This study suggests that an overall reduction in ventilation characterizes oxygen-induced hypercapnia, as an increased dispersion of blood flow from release of hypoxic vasoconstriction occurred to a significant and similar degree in both groups. The significant increase in wasted ventilation (alveolar dead space) in the R group only may be secondary to the higher carbon dioxide tension, perhaps related to bronchodilatation. PMID- 10806150 TI - Implications of extubation delay in brain-injured patients meeting standard weaning criteria. AB - We hypothesized that variation in extubating brain injured patients would affect the incidence of nosocomial pneumonia, length of stay, and hospital charges. In a prospective cohort of consecutive, intubated brain-injured patients, we evaluated daily: intubation status, spontaneous ventilatory parameters, gas exchange, neurologic status, and specific outcomes listed above. Of 136 patients, 99 (73%) were extubated within 48 h of meeting defined readiness criteria. The other 37 patients (27%) remained intubated for a median 3 d (range, 2 to 19). Patients with delayed extubation developed more pneumonias (38 versus 21%, p < 0.05) and had longer intensive care unit (median, 8.6 versus 3.8 d; p < 0.001) and hospital (median, 19.9 versus 13.2 d; p = 0.009) stays. Practice variation existed after stratifying for differences in Glasgow Coma Scale scores (10 versus 7, p < 0.001) at time of meeting readiness criteria, particularly for comatose patients. There was a similar reintubation rate. Median hospital charges were $29,057.00 higher for extubation delay patients (p < 0.001). This study does not support delaying extubating patients when impaired neurologic status is the only concern prolonging intubation. A randomized trial of extubation at the time brain-injured patients fulfill standard weaning criteria is justifiable. PMID- 10806151 TI - Effects of inverse ratio ventilation and positive end-expiratory pressure in oleic acid-induced lung injury. AB - Continuous as well as cyclic (with each expiration) lung collapse in acute respiratory failure can be reduced by positive end-expiratory pressure (PEEP) or short expiration times, as in inverse ratio ventilation (IRV). In 20 pigs with oleic acid-induced lung edema, we compared the effects of a PEEP of 20 cm H(2)O with IRV, using an inspiratory-to-expiratory ratio of 3:1 without external PEEP. During IRV, expiration times of 0.5 or 1.0 s were obtained with respiratory rates of 30 breaths/min or 15 breaths/min, respectively. In 15 animals, ventilation perfusion relationships were studied through the multiple inert gas elimination technique, and lung morphology was studied with computed tomography. In another five pigs, blood flow distribution was studied with perfusion scintigraphy. All three ventilatory modes had similar effects on mean arterial blood pressure, cardiac output, oxygen delivery, and mean airway pressure. PEEP reduced shunt and improved oxygenation to a greater extent than the two modes of IRV, although there was a large variation within each group. The improvement, irrespective of which ventilatory mode was superior in a particular pig, was caused by greater and more even aeration of the lung, whereas the perfusion distribution with PEEP was the same as with IRV. Thus, the strategy of stabilizing the lungs through short expiration times, as in IRV, did not offer any advantages in our lung injury model. PMID- 10806152 TI - Optoelectronic plethysmography in intensive care patients. AB - We used optoelectronic plethysmography to study 11 normal subjects during quiet and deep breathing, six sedated and paralyzed patients with acute lung injury and acute respiratory distress syndrome (ALI/ARDS) receiving continuous positive pressure ventilation (CPPV) (positive end-expiratory pressure [PEEP] = 10 cm H(2)O, tidal volume [VT] = 300, 600, 900 ml), and seven ALI/ARDS patients receiving pressure support ventilation (PSV) (PEEP 10 cm H(2)O, pressure support = 5, 10, 15, 25 cm H(2)O). The volumes measured using optoelectronic plethysmography were compared with measurements taken using spirometry and pneumotachography. The three methods were highly correlated. The discrepancies found were 1.7 +/- 5.9%, -1.6 +/- 5.4%, and 4.9 +/- 6.4% when comparing optoelectronic plethysmography with spirometry, optoelectronic plethysmography with pneumotachography, and spirometry with pneumotachography, respectively. Accuracy of the compartmentalization procedure (upper thorax, lower thorax, and abdomen) was assessed by calculating compartmental volume changes during isovolume maneuvers. The discrepancy from the ideal zero line was -2.1 +/- 48.3 ml. Abdominal contribution to inspired volume was greater for normal subjects than for PSV patients (63 +/- 11% versus 43 +/- 14%, p < 0.001). It decreased with VT for normal subjects (48.5 +/- 15%, p < 0.05), whereas it increased for CPPV patients (61 +/- 10%, p < 0.05). No significant distribution differences were found between 5 and 25 cm H(2)O PSV. We conclude that optoelectronic plethysmography is a feasible technique able to provide unique data on the distribution of chest wall volume changes in intensive care patients. PMID- 10806153 TI - Sputum cysteinyl leukotrienes increase 24 hours after allergen inhalation in atopic asthmatics. AB - We have used the relatively noninvasive technique of induced sputum to measure allergen-induced changes in the concentration of eicosanoid mediators in bronchial secretions from atopic asthmatics. Sputum induction was performed before and 24 h after inhalational allergen challenge in 14 atopic asthmatics who developed a late asthmatic reaction (LAR). Differential cell counts were made on sputum cytospins and eicosanoid (cysteinyl leukotrienes [cys LTs], prostaglandin D(2) [PGD(2)], and PGE(2)) concentrations were measured in the sputum supernatants. The percentage of eosinophils at baseline correlated with the concentration of cys LTs (r = 0.84, p < 0.001) but not prostanoid mediators. Allergen challenge produced a significant increase in the concentration of sputum cys LTs from 3. 45 ng/ml sputum to 11.95 ng/ml (p = 0.002), which correlated with the increase in sputum eosinophils (r = 0.55, p < 0.05). There were no significant changes in PGD(2) or PGE(2) concentrations in sputum supernatants in response to challenge. Thus, the noninvasive technique of induced sputum has been used to demonstrate increased cys LTs, but not prostanoids associated with LAR after allergen challenge. The correlation between eosinophil numbers and cys LT concentrations at baseline values and 24 h after allergen challenge is consistent with these cells being a principal source of cys LTs within the airways at these time points. PMID- 10806154 TI - A minimum 5.0 ml of sputum improves the sensitivity of acid-fast smear for Mycobacterium tuberculosis. AB - Detection of acid-fast bacilli (AFB) by sputum smear supports treatment decisions with pulmonary tuberculosis (TB), but smear sensitivity for Mycobacterium tuberculosis is only approximately 45 to 75%. In an effort to increase sensitivity, smears were prepared using a minimum sputum volume of 5.0 ml. Sensitivity of smears during a 39-mo period (n = 1,849) using >/= 5.0 ml of sputum was 92. 0%, significantly greater (p < 0.001) than a sensitivity of 72.5% in a previous 24-mo period (n = 3,486) when all specimens were processed regardless of volume. All new cases of TB (n = 18) were smear-positive with >/= 5.0 ml of sputum before treatment, and all were receiving antituberculosis drugs at hospital discharge. In contrast, significantly fewer new cases of TB (14 of 26, p = 0.002) were positive before treatment when smears were prepared using sputum of any volume, and significantly fewer of these new TB cases (18 of 26, p = 0.03) were receiving treatment at hospital discharge. The eight cases without treatment were smear-negative. These results indicate that acid-fast smear using >/= 5.0 ml of sputum increases sensitivity for M. tuberculosis and accelerates treatment of TB. PMID- 10806155 TI - Relative scarcity of asthma and atopy among rural adolescents raised on a farm. AB - We determined the prevalence of markers of atopy and asthma among 1, 199 rural secondary school students ages 12 to 19 yr. Subjects identified as having been raised on a farm and half as many subjects without regular exposure to a farming environment from the same school class completed a respiratory symptom questionnaire and underwent allergy skin tests and a methacholine bronchoprovocation test. Current wheeze, airways hyperresponsiveness (AHR), and skin test positivity to inhaled allergens were all significantly less common among adolescents raised on the farm and these differences were especially pronounced in girls. After adjusting for gender and current smoking, the odds ratios for being raised on a farm were: 0. 70 (95% CI 0.52 to 0.95) for current wheeze; 0.59 (95% CI 0.37 to 0. 95) for asthma, defined as the concomitant occurrence of wheeze and AHR; and 0.58 (95% CI 0.46 to 0.75) for atopy defined as a positive reaction to any one of 24 common inhaled allergens. These associations were also not significantly altered by adjusting for the difference in the number of siblings. PMID- 10806156 TI - Thoracoabdominal compression and respiratory system compliance in HIV-infected infants. AB - The thoracoabdominal compression technique (TAC) is used to measure expiratory flow in infants. We investigated whether TAC caused a change in total thoracic compliance (Crs), resistance (Rrs), and respiratory system time constant (Trs). We studied 41 infants (mean age, 12.4 mo; SD, 7.5) from five centers studying longitudinal lung and cardiovascular function of infants from HIV-infected mothers. We measured Crs, Rrs, and Trs before and after TAC. Changes in Crs, Rrs, and Trs after TAC were not dependent on the length of time since TAC. Crs and Trs were reduced after TAC, p = 0.013 and p = 0.003, respectively, whereas Rrs did not change. When compared with uninfected infants, HIV-infected infants had a larger post-pre TAC percent decline in Crs (p = 0.003) and a post-pre TAC rise in mean Rrs (p = 0.03). These differences remained significant after adjusting for sex and age. When performing infant pulmonary function testing, TAC itself produces a temporary decrease in Crs and Trs that is more significant in infants at risk for abnormal lung volume or compliance. Therefore, the sequence of performing the infant lung function parameters should be the same each time the testing is repeated with TAC as the last parameter tested at each testing session. PMID- 10806157 TI - Antituberculosis activity of once-weekly rifapentine-containing regimens in mice. Long-term effectiveness with 6- and 8-month treatment regimens. AB - The effectiveness of various once-weekly 10 mg/kg rifapentine (P)- containing regimens for treatment of tuberculosis was assessed in mice infected intravenously with 4.3 x 10(6) colony-forming units (cfu) of Mycobacterium tuberculosis H37Rv, and treated 14 d later with various combinations of rifampin (R), P, isoniazid (H), pyrazinamide (Z), ethambutol (E), or streptomycin (S). Control mice treated daily with either 2-mo HRZ + 4-mo HR or 2-mo HRZ + 6-mo HE were rendered spleen and lung culture-negative at 6 mo and 8 mo, respectively. Treatment failure with emergence of R-resistant bacilli occurred in all mice given once-weekly monotherapy with P for 6 mo. Once-weekly PH treatment was successful at 6 mo when it was preceded by a 2-mo daily phase with HRZ. When the initial daily phase was reduced to 2 wk, once-weekly PH-containing treatment was successful, at 6 mo, only if it was supplemented with S during the initial daily and the once-weekly phases, and at 8 mo if it was supplemented with daily H during the once-weekly phase. Without these supplements, once-weekly treatment failed in some mice with selection of R-resistant or H-resistant mutants. PMID- 10806158 TI - Factors related to methacholine airway responsiveness in children. AB - Studies of airway responsiveness (AR) have typically used similar dose schedules of methacholine for adults and children despite large ranges in subject size. Reported declines in AR with increasing age in children could be due to maturational changes or to proportionately smaller doses of methacholine in taller (older) children. Other investigators have related both height and various measures of lung function to AR. We examined data related to AR in 471 children, aged 6 to 8 yr, from a birth cohort. Each child underwent spirometry followed by sequential challenge with five doses of methacholine, ranging from 0.025 to 25 mg/ml, given with a dosimeter. Continuous slope and end FEV(1)-change indexes of responsiveness were computed. Using stepwise regression modeling, we found no significant association between AR and either height or age after entering a variable reflecting asthma or wheezing. In contrast, we found that baseline measures of FVC, FEV(1)/FVC, and FEF(25-75%) were significantly related to AR after controlling for other variables (p = 0.001). However, when all three of the latter measures were added to models, FEF(25-75%) was most closely related to AR. We conclude that after control for other variables, FEF(25-75%) and FVC, but not height, are significantly related to methacholine responsiveness in children. PMID- 10806159 TI - Exogenous surfactant kinetics in infant respiratory distress syndrome: A novel method with stable isotopes. AB - Little is known about surfactant metabolism in newborn infants, since radioactive isotopes cannot be used in humans. We describe here a new method for studying exogenous surfactant pharmacokinetics in vivo. We measured surfactant half-life, pool size, and turnover time in eight preterm infants (gestational age: 30 +/- 2 wk; birth weight: 1,416 +/- 202 g) who were mechanically ventilated because of infant respiratory distress syndrome. We administered two doses of 100 mg/kg each of a natural porcine surfactant with (13)C-labeled dipalmitoylphosphatidylcholine as a tracer. The (13)C enrichment of surfactant disaturated phosphatidylcholine (DSPC) was measured in serial tracheal aspirates by gas chromatography-mass spectrometry. The DSPC half-life was 34.2 +/- 9.4 h (mean +/- SD; range: 21.8 to 45.9 h). The apparent DSPC pool sizes were 5.8 +/- 6.1 mg/kg (range: 0.1 to 17.0 mg/kg) and 17.3 +/- 13.6 mg/kg (range: 3.3 to 41.0 mg/kg) at the time of the first and second surfactant doses, respectively. We present a novel and safe method that allows the tracing of exogenous surfactant phosphatidylcholine, the major lipid component of pulmonary surfactant, in infants who receive exogenous surfactant. This method could be a valuable tool for studying: (1) therapies that enhance lung/surfactant maturation; (2) the dosing and timing of surfactant therapy in different lung diseases; and (3) the comparison of different surfactant preparations. PMID- 10806160 TI - Expiratory flow limitation and intrinsic positive end-expiratory pressure at zero positive end-expiratory pressure in patients with adult respiratory distress syndrome. AB - It has been suggested that in patients with adult respiratory distress syndrome (ARDS), intrinsic positive end-expiratory pressure (PEEPi) is generated by a disproportionate increase in expiratory flow resistance. Using the negative expiratory pressure (NEP) technique, we assessed whether expiratory flow limitation (EFL) and PEEPi were present at zero PEEP in 10 semirecumbent, mechanically ventilated ARDS patients. Because bronchodilators may decrease airway resistance, we also investigated the effect of nebulized salbutamol on EFL, PEEPi, and respiratory mechanics in these patients, and in seven patients we measured the latter variables in the supine position as well. In the semirecumbent position, eight of the 10 ARDS patients exhibited tidal EFL, ranging from 5 to 37% of the control tidal volume (VT), whereas PEEPi was present in all 10 subjects, ranging from 0.4 cm H(2)O to 7.7 cm H(2)O. The onset of EFL was heralded by a distinct inflection point on the expiratory flow-volume curve, which probably reflected small-airway closure. Administration of salbutamol had no statistically significant effect on PEEPi, EFL (as %VT), or respiratory mechanics. EFL (%VT) and PEEPi were significantly higher in the supine position than in the semirecumbent position, whereas the other respiratory variables did not change. Our results suggest that in the absence of externally applied PEEP, most ARDS patients exhibit EFL associated with small-airway closure and a concomitant PEEPi. PMID- 10806161 TI - Predictors of mortality in acute respiratory distress syndrome. Focus On the role of right heart catheterization. AB - Right heart catheterization (RHC) has been suspected of increasing mortality. The acute respiratory distress syndrome (ARDS) is a frequent reason for RHC. We designed a retrospective cohort study of 119 consecutive ARDS patients admitted to two medical intensive care units of tertiary care hospitals in which two different approaches are used for hemodynamic monitoring: RHC on demand (Henri Mondor Hospital [HM]) and no use of RHC (Ambroise Pare Hospital [AP]). The study tried to identify risk factors for death, and to assess the influence of RHC, with adjustment for the intensity of hemodynamic support as a confounding factor, using 98 patients in whom the delay between onset of ARDS, use of vasopressors, and RHC did not exceed 48 h. Several variables, including septic shock, cause of ARDS, Simplified Acute Physiology Score (SAPS) II, use of epinephrine/norepinephrine, and presence of RHC were entered into a logistic regression model to evaluate their independent prognostic roles. Mortality was different at HM and AP (36 of 55 patients [65.5%] versus 16 of 43 patients [37.2%], p < 0.005), and 29 of the 35 RHC-monitored patients died (82.8%), as compared with 23 of 63 patients (36.5%) treated without RHC (p < 0.0001). However, administration of epinephrine/norepinephrine and a nonpulmonary cause of ARDS were each independently associated with death. It is only when administration of vasopressors was omitted from the model that RHC, septic shock, and SAPS II became independent predictors of mortality. These results suggest that: (1) the use of vasopressors, but not of RHC, represents an important prognostic factor; and (2) not taking into account the use of these drugs may be misleading when assessing the influence of RHC on outcome. PMID- 10806162 TI - Influence of rheologic changes and platelet-neutrophil interactions on cell filtration in sepsis. AB - We examined the role of erythrocyte (red blood cell; RBC) aggregation and deformability, neutrophil (polymorphonuclear neutrophil; PMN) deformability, whole-blood viscosity, and platelet-neutrophil interactions on cell filtration in subjects who were critically ill with sepsis (CIS), critically ill noninfected subjects (CINS), and healthy controls (C). We assessed cell deformability by filtration through filters of 5-microm pore size. Whole blood, RBC, PMN, and combinations of PMN and RBC were studied. Viscometry was done on isolated RBC. Platelet-PMN interactions were assessed with monoclonal antibodies to CD41 and activated CD63 platelet receptors, and to CD66b PMN receptors. Filtration pressure (Pi) for CIS was significantly greater than for C and CINS at both high and low PMN and RBC concentrations. Viscometry confirmed decreases in RBC deformability and demonstrated significant increases in RBC aggregation in CIS. Increments in Pi were significantly greater with PMN and PMN-RBC combinations suspended in platelet rich plasma (PRP) than in platelet poor plasma (PPP) for CIS as compared with CINS or C. Flow cytometry confirmed significantly greater platelet activation in CIS than in CINS or C (mean fluorescence: 39 +/- 9 lfu versus 18.7 +/- 4.0 lfu and 17.1 +/- 2.3 lfu, respectively) and greater platelet PMN aggregation (mean fluorescence: 44.7 +/- 3.6 lfu versus 23 +/- 4.1 lfu, respectively) in CIS than in C. We conclude that decreased filtration of whole blood in CIS is related to decreases in RBC and PMN deformability, increases in RBC aggregation, and increased platelet-PMN interactions. Of these, the formation of platelet-PMN aggregates appeares to have the greatest effect in impairing cell filtration. These rheologic abnormalities may contribute to impaired microvascular blood flow in patients with sepsis. PMID- 10806163 TI - Time course and recovery of exacerbations in patients with chronic obstructive pulmonary disease. AB - Although exacerbations of chronic obstructive pulmonary disease (COPD) are associated with symptomatic and physiological deterioration, little is known of the time course and duration of these changes. We have studied symptoms and lung function changes associated with COPD exacerbations to determine factors affecting recovery from exacerbation. A cohort of 101 patients with moderate to severe COPD (mean FEV(1) 41.9% predicted) were studied over a period of 2.5 yr and regularly followed when stable and during 504 exacerbations. Patients recorded daily morning peak expiratory flow rate (PEFR) and changes in respiratory symptoms on diary cards. A subgroup of 34 patients also recorded daily spirometry. Exacerbations were defined by major symptoms (increased dyspnea, increased sputum purulence, increased sputum volume) and minor symptoms. Before onset of exacerbation there was deterioration in the symptoms of dyspnea, sore throat, cough, and symptoms of a common cold (all p < 0.05), but not lung function. Larger falls in PEFR were associated with symptoms of increased dyspnea (p = 0.014), colds (p = 0.047), or increased wheeze (p = 0.009) at exacerbation. Median recovery times were 6 (interquartile range [IQR] 1 to 14) d for PEFR and 7 (IQR 4 to 14) d for daily total symptom score. Recovery of PEFR to baseline values was complete in only 75.2% of exacerbations at 35 d, whereas in 7.1% of exacerbations at 91 d PEFR recovery had not occurred. In the 404 exacerbations where recovery of PEFR to baseline values was complete at 91 d, increased dyspnea and colds at onset of exacerbation were associated with prolonged recovery times (p < 0.001 in both cases). Symptom changes during exacerbation do not closely reflect those of lung function, but their increase may predict exacerbation, with dyspnea or colds characterizing the more severe. Recovery is incomplete in a significant proportion of COPD exacerbations. PMID- 10806164 TI - Overcoming the adverse effect of humidity in aerosol delivery via pressurized metered-dose inhalers during mechanical ventilation. AB - The well-known problem of reduced drug delivery that occurs when heated, humid air is used with pressurized metered-dose inhalers (pMDIs) and spacers in intubated settings is carefully studied with Airomir using an in vitro model under controlled conditions of temperature and humidity. A better understanding of the physical processes leading to the aforementioned drop in performance is obtained, and a method is devised to circumvent the problem without having to reduce the temperature or humidity of the ventilator circuit. The present study shows that the mole fraction of water vapor in the ventilation air (and not the temperature) is the major factor behind the sharp drop in the amount of drug delivered to the lung. However, the presence of water vapor does not affect performance because of hygroscopic growth. Instead, it influences the initial atomization process and the early stages of aerosol generation. Removal of these negative effects can be achieved by using a larger spacer that allows longer times for the aerosol to evaporate, as is demonstrated in the present study. PMID- 10806165 TI - Potential mechanism of hyperresponsive airways. AB - It has been known for many years that the response of asthmatic subjects to a deep inspiration differs from that observed in normal healthy subjects. A deep inspiration causes a decrease in airway resistance in normal subjects, whereas asthmatics demonstrate either no change or a slight increase in airway resistance. It has been suggested by several investigators that the inability to dilate airways during lung inflation may be a primary defect in asthma. One study (Skloot and colleagues, J. Clin. Invest. 1995;96:2393- 2403) showed that in the absence of a deep inspiration during methacholine (MCh) challenge, normal subjects had a greatly exaggerated and sustained response to this agonist. It was suggested that asthmatic airways could be modeled by this condition in normal, subjects. Other investigators, however, suggest that there are more intrinsic differences between the responses to lung inflation in airways from asthmatic and normal subjects (Brusasco and colleagues, J. Appl. Physiol. 1999;87:567-573). Resolution of this controversy requires the ability to assess the responses of airways directly, but unfortunately conventional pulmonary function tests in human subjects are not specific enough to allow this evaluation. In the present study, we have performed experiments using a direct imaging approach that allows us to obtain measurements of airway and parenchymal dimensions that can be used to test the responses of individual airways to deep inspiration in vivo. Our results show that the presence of normal tidal stresses allows airway smooth muscle to respond normally to deep inspirations. Removing tidal stresses at FRC after MCh challenge is sufficient to change the normal dilatory response to deep inspiration into an abnormal one of contraction. Altered sensitivity of airway smooth muscle to normal tidal stresses thus may be operative in the development of the asthmatic pathology. PMID- 10806166 TI - Clara cell specific protein (CC16) expression after acute lung inflammation induced by intratracheal lipopolysaccharide administration. AB - Clara cell secretory protein (CC16, CC10, or CCSP), the major secretory protein of the Clara cell, presents several biologic properties, suggesting that it may play a protective role against intrapulmonary inflammatory processes. The aim of the present study was to investigate the changes of CC16 concentrations in the lung, bronchoalveolar lavage fluid (BALF), and serum of rats with acute lung injury induced by lipopolysaccharide (LPS). These changes were compared with Clara cell density, CC16 mRNA level in the lung and classic indices of inflammation in BALF. Injected at doses of 10, 100, or 200 microgram/100 g body weight, LPS induced an acute lung inflammation as estimated by an increased influx of cells and albumin in the BALF. This inflammatory response was associated with a marked reduction of CC16 concentrations in BALF and lung homogenate as well as of the CC16 mRNA levels in the lung. At the highest dose of LPS, the CC16-positive cell density in the bronchiolar epithelium was also decreased. In serum, by contrast, the concentration of CC16 was elevated as a consequence of increased airway permeability. Pretreating rats intraperitoneally with dexamethasone (2 mg/kg) significantly lowered the leukocyte influx and attenuated the albumin increase in BALF. Dexamethasone, however, failed to prevent the increased airway permeability to CC16, suggesting that during inflammation different mechanisms regulate the leakage of proteins across the alveolocapillary barrier depending on the direction of passage and/or the size of the protein. Our results show a marked decrease of the secretion and synthesis of CC16 during LPS-induced acute lung inflammation. PMID- 10806167 TI - Nerve growth factor expression and release in allergic inflammatory disease of the upper airways. AB - It is well known that allergic airways disease is characterized by inflammation and hyperresponsiveness, but the link between these two conditions has not been elucidated. We have previously shown that in allergic rhinitis, hyperresponsiveness is attributable to increased neural reactivity. We thus hypothesized that nerve growth factor (NGF), which is expressed by inflammatory cells and effects changes that lead to increased neural responsiveness, could be a pivotal mediator in this disease. Using reverse transcription-polymerase chain reaction (RT-PCR), Western immunoblotting, and ELISA to evaluate NGF expression and release, we found that subjects with allergic rhinitis have significantly decreased NGF mRNA in superficial nasal scrapings and significantly higher baseline concentrations of NGF protein in nasal lavage fluids, compared with control subjects. Nasal provocation with allergen significantly increased NGF protein in nasal lavage fluids of subjects with allergic rhinitis, but not of control subjects. The concentrations of NGF protein in nasal lavage fluids were not affected by provocation with the vehicle for allergen or with histamine. These data provide the first evidence of a steady state of dysregulation in mucosal NGF expression and release in allergic rhinitis, and support a role of this neurotrophin in the pathophysiology of allergic inflammatory disease of the human airways. PMID- 10806168 TI - CD13/aminopeptidase N, a novel chemoattractant for T lymphocytes in pulmonary sarcoidosis. AB - CD13/aminopeptidase N (E.C.3.4.11.2) is an ectoenzyme located in the outer membrane of a variety of cells. Because aminopeptidase expression was shown to be upregulated by a Th1-related cytokine, IFN-gamma, we examined here the significance of CD13/aminopeptidase N in pulmonary sarcoidosis. The activity of aminopeptidase in bronchoalveolar lavage fluid (BALF) was significantly higher in patients with sarcoidosis than in normal volunteers (NV) and control patients (CP). The activity significantly correlated with lymphocyte percentages and the ratio of CD4+ to CD8+ T lymphocytes in the BALF, and was higher in patients with sarcoidosis with parenchymal involvement than in those without the involvement. CD13/aminopeptidase N protein, which has a molecular mass of approximately 150 kD, was detectable in alveolar macrophages (AM) from patients with sarcoidosis at higher levels than in those from NV. CD13/aminopeptidase N induced in vitro chemotactic migration of human lymphocytes in a concentration range of 10(-)(5) to 10(-)(1) U/ml. The chemotactic activity was greater for CD4+ T lymphocytes than for CD8+ T lymphocytes. The enzymatic activity of CD13/aminopeptidase N was responsible for the chemotactic activity because bestatin, an inhibitor of CD13/aminopeptidase N, abolished the chemotactic activity. Higher chemotactic activity for lymphocytes was detected in the BALF from patients with sarcoidosis than in that from NV, and the activity was significantly decreased by treatment with bestatin. This study indicates that CD13/ aminopeptidase N expressed in AM may have a role in T-lymphocyte involvement in the sarcoid lung and the pathogenesis of alveolitis in this disorder. PMID- 10806169 TI - Familial pulmonary Mycobacterium avium complex disease. AB - We report two Japanese families affected by pulmonary Mycobacterium avium complex (MAC) disease, involving an older brother and younger sister in one family and two brothers in the second family. We investigated whether defects in the natural resistance-associated macrophage protein gene (NRAMP1) underlay susceptibility to MAC in these cases. All of the patients had computed tomographic findings of peripheral nodules and bronchiectasis. Pulse-field gel electrophoresis patterns of mycobacterial genomic DNA restriction fragments revealed that none of the MAC strains isolated from the patients was epidemiologically related to any of the others. Direct sequencing of the complementary DNA of the patients' NRAMP1 revealed a nonconservative missense mutation at codon 419 in one patient, which was heterozygous and was not seen in his affected sibling. No variations similar to those found in mice that show susceptibility to MAC were found. The results suggest an underlying genetic defect in host defense rather than exposure to an unusually virulent strain of MAC as the pathogenetic factor in MAC disease; however, alterations in the coding region of NRAMP1 do not appear to explain the susceptibility to MAC. PMID- 10806170 TI - A mechanism of antigen-induced mucus production in nasal epithelium of sensitized rats. A comparison with lipopolysaccharide-induced mucus production. AB - We produced ovalbumin (OVA)-sensitized rats as an animal model of nasal allergy. Intranasal instillation of OVA induced hypertrophic and metaplastic changes of goblet cells in nasal epithelium of OVA- sensitized rats. Intraepithelial mucosubstance in nasal mucosa increased significantly at 24 h after 3 or 7 d of OVA instillation, accompanied by mucosal infiltration of eosinophils. The effects of H1-antagonist (d-chlorpheniramine malate), H2-antagonist (cimetidine), dexamethasone, indomethacin, cysteinyl leukotrienes (cysLTs)-antagonist (ONO1078), and antirat neutrophil antiserum on OVA-induced changes were examined. Mucus production was significantly inhibited by dexamethasone, and ONO1078, whereas eosinophil infiltration was significantly inhibited by H1-antagonist, dexamethasone, and anti-rat neutrophil antiserum. These results indicate that cysLTs (LTs C4, D4, and E4) may play an important role in antigen-induced mucus production, and that eosinophil infiltration does not relate to mucus production. Intranasal instillation of lipopolysaccharide (LPS) also induced intraepithelial mucus production, and it was significantly inhibited by dexamethasone, indomethacin, and antirat neutrophil antiserum; however, cysLTs antagonist had no effect on LPS-induced change. These results indicate that neutrophil and cyclooxygenase products are important in LPS-induced mucus production, and there are different mechanisms of mucus production between allergic inflammation and LPS stimulation. PMID- 10806171 TI - Polymorphisms of the IL-4, TNF-alpha, and Fcepsilon RIbeta genes and the risk of allergic disorders in at-risk infants. AB - Polymorphisms in the TNF-alpha (A-308G), IL-4 (C-589T), and Fcalpha RIbeta (E237G) genes have been associated with asthma and related phenotypes. To determine the predictive value of these polymorphisms we have assessed their relative risk (RR) for the development of atopy, asthma, and rhinitis in a high risk infant population that is being followed longitudinally from birth. DNA was extracted and genotyped for 373 infants and 572 parents for each polymorphism. Phenotypic data were collected for atopy and allergic diseases in the infants at 12 mo of age. The prevalence of these phenotypes in the 281 white infants was compared in each genotypic group. There were no differences in the prevalence of any phenotype between genotypes of the TNF-alpha and Fcalpha RIbeta polymorphisms. However, we found that the IL4-589*T allele was associated with "probable" asthma (RR = 4.1) and that homozygotes for the IL4-589*T allele had an increased risk for the development of rhinitis (RR = 2.4). Using the transmission disequilibrium test, an association of IL4-589*T with atopy was found. We conclude that IL-4-589*T, but not TNF-alpha-308*2 or Fcalpha RIbeta*G, is a risk factor for the development of atopy, asthma, and rhinitis by 12 mo of age. PMID- 10806172 TI - The prone position eliminates compression of the lungs by the heart. AB - The prone position improves gas exchange in many patients with ARDS. Animal studies have indicated that turning prone restores ventilation to dorsal lung regions without markedly compromising ventral regions. To investigate a potential mechanism by which this might occur, the relative volume of lung located directly under the heart was measured in the supine and prone positions in seven patients. Four axial tomographic sections between the carina and the diaphragm were analyzed (Sections 1 through 4). When supine, the percent of the total lung volume located under the heart increased from 7 +/- 4% to 42 +/- 8%, and from 11 +/- 4% to 16 +/- 4% in Sections 1 through 4, in the left and right lungs, respectively. When prone, the percent of left and right lung volume located under the heart was /= 50 ppb. Exhaled NO levels (geometric mean) were 15.5 ppb (95% CI: 10.5 to 22.9 ppb) for children from homes with formaldehyde concentrations >/= 50 ppb compared with 8.7 ppb (7.9 to 9.6) for children from homes with formaldehyde concentrations < 50 ppb (p < 0.05). These results suggest that exposure to formaldehyde in homes may invoke a subclinical inflammatory response in the airways of healthy children. PMID- 10806185 TI - The raised volume rapid thoracoabdominal compression technique. The Joint American Thoracic Society/European Respiratory Society Working Group on Infant Lung Function. PMID- 10806186 TI - Heart catheterization in acute lung injury:an observational study. PMID- 10806187 TI - Eosinophilic bronchitis is an important cause of chronic cough. PMID- 10806188 TI - Drug-stimulated ATPase activity of human P-glycoprotein is blocked by disulfide cross-linking between the nucleotide-binding sites. AB - P-glycoprotein (P-gp) is an ATP-dependent drug pump that contains two nucleotide binding domains (NBDs). Disulfide cross-linking analysis was done to determine if the two NBDs are close to each other. Residues within or close to the Walker A (GNSGCGKS in NDB1 and GSSGCGKS in NBD2) sequences for nucleotide binding were replaced with cysteine, and the mutant P-gps were subjected to oxidative cross linking. Cross-linking was detected in two mutants, G427C(NBD1)/Cys-1074(NBD2) and L439C(NBD1)/Cys-1074(NBD2), because the cross-linked proteins migrated slower in SDS gels. Mutants G427C(NBD1)/Cys-1074(NBD2) and L439C(NBD1)/Cys-1074(NBD2) retained 10% and 82%, respectively, of the drug-stimulated ATPase activity relative to that of Cys-less P-gp. The cross-linking properties of the more active mutant L439C(NBD1)/Cys-1074(NBD2) were then studied. Cross-linking was reversed by addition of dithiothreitol and could be prevented by pretreatment of the mutant with N-ethylmaleimide. Cross-linking was also inhibited by MgATP, but not by the verapamil. Oxidative cross-linking of mutant L439C(NBD1)/Cys 1074(NBD2) resulted in almost complete inhibition of drug-stimulated ATPase activity. More than 60% of the drug-stimulated ATPase activity, however, was recovered after treatment with dithiothreitol. The results indicate that the two predicted nucleotide-binding sites are close to each other and that cross-linking inhibits ATP hydrolysis. PMID- 10806189 TI - The mechanism of insulin resistance caused by HIV protease inhibitor therapy. AB - Retroviral protease inhibitors used as therapy for HIV-1 infection have been causally associated with serious metabolic side effects, including peripheral lipodystrophy, hyperlipidemia, insulin resistance, and in some cases, overt type 2 diabetes. The etiology of this characteristic clinical syndrome remains unknown. We demonstrate that the HIV protease inhibitor, indinavir, dramatically inhibits insulin-stimulated glucose uptake in 3T3-L1 adipocytes in a dose dependent manner (63% inhibition observed with 100 micrometer indinavir). Indinavir treatment did not affect early insulin signaling events or the translocation of intracellular Glut1 or Glut4 glucose transporters to the cell surface. To determine whether indinavir may be directly affecting the intrinsic transport activity of glucose transporters, the Glut1 and Glut4 isoforms were heterologously expressed and analyzed in Xenopus laevis oocytes. Indinavir at 100 microm had no effect on Glut1 transport activity in Xenopus oocytes, whereas Glut4 activity was significantly inhibited (45% inhibition). Similar effects on glucose transport were observed for other HIV protease inhibitors. We conclude that HIV protease inhibitors as a class are capable of selectively inhibiting the transport function of Glut4 and that this effect may be responsible for a major iatrogenic complication frequently observed in HIV patients. PMID- 10806190 TI - Covalent modification of the Werner's syndrome gene product with the ubiquitin related protein, SUMO-1. AB - Werner's syndrome is a potential model of accelerated human aging. The gene responsible for Werner's syndrome encodes a protein that has a helicase domain homologous to Escherichia coli RecQ. To identify binding partners that regulate the function in concert with Wrn, we screened for proteins using the yeast two hybrid system with mouse Wrn as bait and found three. One was a novel protein, and the other two were mouse Ubc9 and SUMO-1. Ubc9 also interacted with the mouse homologue of the Bloom's syndrome gene product, another eukaryotic RecQ-type helicase, but not mouse DNA helicase Q1/RecQL (RecQL1). Deletion experiments indicated that both proteins interacted with the N-terminal segment of Wrn (amino acid 272-514). The interaction between Wrn and SUMO-1 was weaker than that between Wrn and Ubc9. Positive interaction was observed in the heterogeneous combination of Wrn and yeast Ubc9 (yUbc9), as well as yUbc9 and SUMO-1, in the two-hybrid system. The interaction between yUbc9 and SUMO-1 was abolished by deleting the C-terminal Gly residue of SUMO-1, which is reportedly required for the formation of Ubc9-SUMO-1 thioester linkage. The interaction of Wrn and SUMO-1 was also abolished by deleting the Gly residue, indicating that the interaction of Wrn and SUMO-1 is mediated by yUbc9 in the two-hybrid system. Finally, we confirmed by immunoblotting with an anti-SUMO-1 antibody that Wrn was covalently attached with SUMO-1. PMID- 10806191 TI - A tuftelin-interacting protein (TIP39) localizes to the apical secretory pole of mouse ameloblasts. AB - Enamel biomineralization is a complex process that involves interactions between extracellular matrix proteins. To identify proteins interacting with tuftelin, a potential nucleator of enamel crystallites, the yeast two-hybrid system was applied to a mouse tooth expression library and a tuftelin-interacting protein (TIP) was isolated for further characterization. Polyclonal antibodies were prepared against two recombinant variants of this protein. Both antibodies identified a major protein product in tooth organs at 39 kDa, and this protein has been called TIP39. Northern analysis showed TIP39 messenger RNA in multiple organs, a pattern similar to that of tuftelin messenger RNA. In situ hybridization of mandibles of 1-day-old mice detected TIP39 RNA in secretory ameloblasts and odontoblasts. Immunolocalization of TIP39 and tuftelin in cultured ameloblast-like cells showed that these two proteins colocalize. Within the developing tooth organ, TIP39 and tuftelin immunolocalized to the apical pole of secretory ameloblasts (Tomes' processes) and to the newly secreted extracellular enamel matrix. TIP39 amino acid sequence appears to be highly conserved with similarities to proteins in species as diverse as yeast and primates. Available sequence data and the findings reported here suggest a role for TIP39 in the secretory pathway of extracellular proteins. PMID- 10806192 TI - Pigment binding site properties of two photosystem II antenna proteins. A resonance raman investigation. AB - Two light-harvesting proteins associated with photosystem II of higher plants, namely the major antenna complex LHCIIb and the minor Lhcb4 protein (CP29), have been investigated by resonance Raman spectroscopy. One of the two chlorophylls b and up to five of the six chlorophylls a present in Lhcb4 are shown to adopt similar binding conformations to the (presumably) corresponding molecules in LHCIIb, whereas at least two chlorophylls in the former protein assume unique conformations relative to the bulk complex. The overall conformation of bound xanthophyll molecules is identical in the two antenna proteins, although some small differences are apparent. The pigment binding properties of these two LHCs are discussed, with particular reference to possible structural motifs within this extended family of proteins. PMID- 10806193 TI - A conformation change in the carboxyl terminus of Alzheimer's Abeta (1-40) accompanies the transition from dimer to fibril as revealed by fluorescence quenching analysis. AB - Alzheimer's disease is characterized by the presence of insoluble, fibrous deposits composed principally of amyloid beta (Abeta) peptide. A number of studies have provided information on the fibril structure and on the factors affecting fiber formation, but the details of the fibril structure are not known. We used fluorescence quenching to investigate the solvent accessibility and surface charge of the soluble Abeta(1-40) dimer and amyloid fibrils. Analogs of Abeta(1-40) containing a single tryptophan were synthesized by substituting residues at positions 4, 10, 34, and 40 with tryptophan. Quenching measurements in the dimeric state indicate that the amino-terminal analogs (AbetaF4W and AbetaY10W) are accessible to polar quenchers, and the more carboxyl-terminal analog AbetaV34W is less accessible. AbetaV40W, on the other hand, exhibits a low degree of quenching, indicating that this residue is highly shielded from the solvent in the dimeric state. Correcting for the effect of reduced translational and rotational diffusion, fibril formation was associated with a selective increase in solvent exposure of residues 34 and 40, suggesting that a conformation change may take place in the carboxyl-terminal region coincident with the dimer to fibril transition. PMID- 10806194 TI - Cloning and characterization of a 72-kDa inositol-polyphosphate 5-phosphatase localized to the Golgi network. AB - The inositol-polyphosphate 5-phosphatase enzyme family removes the 5-position phosphate from both inositol phosphate and phosphoinositide signaling molecules. We have cloned and characterized a novel 5-phosphatase, which demonstrates a restricted substrate specificity and tissue expression. The 3.9-kb cDNA predicts for a 72-kDa protein with an N-terminal proline rich domain, a central 5 phosphatase domain, and a C-terminal CAAX motif. The 3. 9-kilobase mRNA showed a restricted expression but was abundant in testis and brain. Antibodies against the sequence detected a 72-kDa protein in the testis in the detergent-insoluble fraction. Indirect immunofluorescence of the Tera-1 cell line using anti-peptide antibodies to the 72-kDa 5-phosphatase demonstrated that the enzyme is predominantly located to the Golgi. Expression of green fluorescent protein tagged 72-kDa 5-phosphatase in COS-7 cells revealed that the enzyme localized predominantly to the Golgi, mediated by the N-terminal proline-rich domain, but not the C-terminal CAAX motif. In vitro, the protein inserted into microsomal membranes on the cytoplasmic face of the membrane. Immunoprecipitated recombinant 72-kDa 5-phosphatase hydrolyzed phosphatidylinositol 3,4,5-trisphosphate and phosphatidylinositol 3, 5-bisphosphate, forming phosphatidylinositol 3,4 bisphosphate and phosphatidylinositol 3-phosphate, respectively. We propose that the novel 5-phosphatase hydrolyzes phosphatidylinositol 3,4, 5-trisphosphate and phosphatidylinositol 3,5-bisphosphate on the cytoplasmic Golgi membrane and thereby may regulate Golgi-vesicular trafficking. PMID- 10806195 TI - Cloning of two human thyroid cDNAs encoding new members of the NADPH oxidase family. AB - Two cDNAs encoding NADPH oxidases and constituting the thyroid H(2)O(2) generating system have been cloned. The strategy of cloning was based on the functional similarities between H(2)O(2) generation in leukocytes and the thyroid, according to the hypothesis that one of the components of the thyroid system would belong to the gp91(Phox)/Mox1 gene family and display sequence similarities with gp91(Phox). Screening at low stringency with a gp91(Phox) probe of cDNA libraries from thyroid cells in primary culture yielded two distinct human cDNA clones harboring open reading frames of 1551 (ThOX1) and 1548 amino acids (ThOX2), respectively. The encoded polypeptides display 83% sequence similarity and are clearly related to gp91(Phox) (53 and 47% similarity). The theoretical molecular mass of 177 kDa is close to the apparent molecular mass of 180 kDa of the native corresponding porcine flavoprotein and the protein(s) detected by Western blot in dog and human thyroid. ThOX1 and ThOX2 display sequence similarities of 53% and 61%, respectively, with a predicted protein of Caenorhabditis elegans over their entire length. They show along their first 500 amino acids a similarity of 43% with thyroperoxidase. The corresponding genes of ThOX1 and ThOX2 are closely linked on chromosome 15q15.3. The dog mRNA expression is thyroid-specific and up-regulated by agents activating the cAMP pathway as is the synthesis of the polypeptides they are coding for. In human thyroid the positive regulation by cAMP is less pronounced. The proteins ThOX1 and ThOX2 accumulate at the apical membrane of thyrocytes and are co-localized with thyroperoxidase. PMID- 10806197 TI - Phosphorylation of human glutamine:fructose-6-phosphate amidotransferase by cAMP dependent protein kinase at serine 205 blocks the enzyme activity. AB - Glutamine:fructose-6-phosphate amidotransferase (GFAT) is the rate-limiting enzyme in glucosamine synthesis. Prior studies from our laboratory indicated that activation of adenylate cyclase was associated with depletion of O-GlcNAc modification. This finding and evidence that human GFAT (hGFAT) might be regulated by cAMP-dependent protein kinase (PKA) led us to investigate the role of PKA in hGFAT function. We confirmed that adenylate cyclase activation by forskolin results in diminished O-GlcNAc modification of several cellular proteins which can be overcome by exposure of the cells to glucosamine but not glucose, suggesting the PKA activation results in depletion of UDP-GlcNAc for O glycosylation. To determine if GFAT is indeed regulated by PKA, we expressed the active form of the enzyme using a vaccinia virus expression system and showed that the activity of the enzyme was to decrease to undetectable levels by PKA phosphorylation. We mapped the PKA phosphorylation sites with the aid of matrix assisted laser desorption ionization mass spectroscopy and showed that the protein was stoichiometrically phosphorylated at serine 205 and also phosphorylated, to a lesser extent at serine 235. Mutagenesis studies indicated that the phosphorylation of serine 205 by PKA was necessary for the observed inhibition of enzyme activity while serine 235 phosphorylation played no observable role. The activity of GFAT is down-regulated by cAMP, thus placing regulation on the hexosamine pathway that is in concert with the energy requirements of the organism. During starvation, hormones acting through adenylate cyclase could direct the flux of glucose metabolism into energy production rather than into synthetic pathways that require hexosamines. PMID- 10806198 TI - Biochemical evidence for the involvement of tyrosine in epoxide activation during the catalytic cycle of epoxide hydrolase. AB - Epoxide hydrolases (EH) catalyze the hydrolysis of epoxides and arene oxides to their corresponding diols. The crystal structure of murine soluble EH suggests that Tyr(465) and Tyr(381) act as acid catalysts, activating the epoxide ring and facilitating the formation of a covalent intermediate between the epoxide and the enzyme. To explore the role of these two residues, mutant enzymes were produced and the mechanism of action was analyzed. Enzyme assays on a series of substrates confirm that both Tyr(465) and Tyr(381) are required for full catalytic activity. The kinetics of chalcone oxide hydrolysis show that mutation of Tyr(465) and Tyr(381) decreases the rate of binding and the formation of an intermediate, suggesting that both tyrosines polarize the epoxide moiety to facilitate ring opening. These two tyrosines are, however, not implicated in the hydrolysis of the covalent intermediate. Sequence comparisons showed that Tyr(465) is conserved in microsomal EHs. The substitution of analogous Tyr(374) with phenylalanine in the human microsomal EH dramatically decreases the rate of hydrolysis of cis stilbene oxide. These results suggest that these tyrosines perform a significant mechanistic role in the substrate activation by EHs. PMID- 10806199 TI - Granulocyte colony-stimulating factor induces Egr-1 up-regulation through interaction of serum response element-binding proteins. AB - Granulocyte colony-stimulating factor (G-CSF) stimulates the proliferation and maturation of myeloid progenitor cells both in vitro and in vivo. We showed that G-CSF rapidly and transiently induces expression of egr-1 in the NFS60 myeloid cell line. Transient transfections of NFS60 cells with recombinant constructs containing various deletions of the human egr-1 promoter identified the serum response element (SRE) between nucleotides (nt) -418 and -391 as a critical G-CSF responsive sequence. The SRE (SRE-1) contains a CArG box, the binding site for the serum response factor (SRF), which is flanked at either side by an ETS protein binding site. We demonstrated that a single copy of the wild-type SRE-1 in the minimal promoter plasmid, pTE2, is sufficient to induce transcriptional activation in response to G-CSF and that both the ETS protein binding site and the CArG box are required for maximal transcriptional activation of the pTE2-SRE 1 construct. In electromobility shift assays using NFS60 nuclear extracts, we identified SRF and the ETS protein Fli-1 as proteins that bind the SRE-1. We also demonstrated through electrophoretic mobility shift assays, using an SRE-1 probe containing a CArG mutation, that Fli-1 binds the SRE-1 independently of SRF. Our data suggest that SRE-binding proteins potentially play a role in G-CSF-induced egr-1 expression in myeloid cells. PMID- 10806200 TI - Intracellular retention and degradation of the epidermal growth factor receptor, two distinct processes mediated by benzoquinone ansamycins. AB - Epidermal growth factor (EGF) stimulates the growth of various types of cells via its cell surface tyrosine kinase receptor. The EGF receptor (EGF-R) has an oncogenic potential when overexpressed in a wide range of tumor cells. Geldanamycin (GA) and herbimycin (HA), specific inhibitors of the cytosolic chaperone HSP 90 and its endoplasmic reticulum homologue GRP 94, were shown to accelerate degradation of the EGF-R and of its homologue p185(c-)(erbB-2). Here we compared the effects of GA and HA on intracellular degradation and maturation of EGF-R. By using an inhibitor of proteasomal degradation, we learned that GA, but not HA, blocks processing of newly synthesized EGF-R. The effects of GA and HA on receptor degradation are mediated by the cytosolic portion of EGF-R and could be conferred to the erythropoietin receptor (EPO-R), by employing the respective chimera. Neither HA nor GA affected stability of newly synthesized EGF R lacking the cytosolic domain (Ex EGF-R), but GA caused intracellular retention of this mutant. Taken together, our results imply that GA has two distinct targets of action on the EGF-R, one for promoting its degradation and another for mediating its intracellular retention. Apparently, degradation of the EGF-R mediated by GA or HA requires the presence of the EGF-R cytosolic domain, whereas intracellular retention in the presence of GA is coupled to the extracellular domain of the EGF-R. PMID- 10806201 TI - Age-related decline in chaperone-mediated autophagy. AB - Intracellular protein degradation rates decrease with age in many tissues and organs. In cultured cells, chaperone-mediated autophagy, which is responsible for the selective degradation of cytosolic proteins in lysosomes, decreases with age. In this work we use lysosomes isolated from rat liver to analyze age-related changes in the levels and activities of the main components of chaperone-mediated autophagy. Lysosomes from "old" (22-month-old) rats show lower rates of chaperone mediated autophagy, and both substrate binding to the lysosomal membrane and transport into lysosomes decline with age. A progressive age-related decrease in the levels of the lysosome-associated membrane protein type 2a that acts as a receptor for chaperone-mediated autophagy was responsible for decreased substrate binding in lysosomes from old rats as well as from late passage human fibroblasts. The cytosolic levels and activity of the 73-kDa heat-shock cognate protein required for substrate targeting to lysosomes were unchanged with age. The levels of lysosome-associated hsc73 were increased only in the oldest rats. This increase may be an attempt to compensate for reduced activity of the pathway with age. PMID- 10806202 TI - Quantitative analysis of nuclear localization signal (NLS)-importin alpha interaction through fluorescence depolarization. Evidence for auto-inhibitory regulation of NLS binding. AB - We have developed a quantitative in vitro steady-state fluorescence depolarization assay to measure the interaction of a nuclear localization signal (NLS) substrate with its receptors. This assay relies on the change in fluorescence depolarization of an NLS fused to the green fluorescent protein upon binding to receptor. No binding is observed in the absence of a functional NLS, and binding affinities measured correlate with previous in vivo studies of NLS function. We have used this assay to test an auto-inhibitory model for the interaction of an NLS with the NLS receptor complex. This model suggests that NLS binding to importin alpha is modulated by an auto-inhibitory sequence within the N terminus of importin alpha, which is displaced by importin beta binding. Consistent with this model, NLS substrates bind tightly to an N-terminally truncated importin alpha lacking the auto-inhibitory domain (K(d) approximately 10 nm), but measurable binding to full-length importin alpha is only observed upon addition of importin beta. Our quantitative results support the auto inhibitory model and suggest a mechanism for a switch between a cytoplasmic, high affinity and a nuclear, low affinity NLS receptor. This predicted mode of interaction would facilitate binding of substrate in the cytoplasm and its subsequent release into the nucleus. PMID- 10806203 TI - Bone morphogenetic protein 1 is an extracellular processing enzyme of the laminin 5 gamma 2 chain. AB - Epithelial cells maintained in culture medium containing low calcium proteolytically process laminin 5 (alpha3beta3gamma2) within the alpha3 and gamma2 chains (). Experiments were designed to identify the enzyme(s) responsible for the laminin 5 processing and the sites of proteolytic cleavage. To characterize the nature of laminin 5 processing, we determined the N-terminal amino acid sequences of the proteolytic fragments produced by the processing events. The results indicate that the first alpha3 chain cleavage (200-l65 kDa alpha3) occurs within subdomain G4 of the G domain. The second cleavage (l65-l45 kDa alpha3) occurs within the lIla domain, 11 residues N-terminal to the start of domain II. The gamma chain is cleaved within the second epidermal growth factor like repeat of domain Ill. The sequence cleaved within the gamma2 chain matches the consensus sequence for the cleavage of type I, II, and III procollagens by bone morphogenetic protein-1 (BMP-1), also known as type I procollagen C proteinase (). Recombinant BMP-1 cleaves gamma2 in vitro, both within intact laminin 5 and at the predicted site of a recombinant gamma2 short arm. alpha3 is also cleaved by BMP-1 in vitro, but the cleavage site is yet to be determined. These results show the laminin alpha3 and gamma2 chains to be substrates for BMP 1 in vitro. We speculate that gamma2 cleavage is required for formation of the laminin 5-6 complex and that this complex is directly involved in assembly of the interhemidesmosomal basement membrane. This further suggests that BMP-1 activity facilitates basement membrane assembly, but not hemidesmosome assembly, in the laminin 5-rich dermal-epidermal junction basement membrane in vivo. PMID- 10806205 TI - Altered regulatory properties of human cardiac troponin I mutants that cause hypertrophic cardiomyopathy. AB - Cardiac troponin I (cTnI) is the inhibitory component of the troponin complex and is involved in the calcium control of heart muscle contraction. Recently, specific missense mutations of the cTnI gene (TNNI3) have been shown to cause familial hypertrophic cardiomyopathy (HCM). We have analyzed the functional effects of two HCM mutations (R145G and R162W) using purified recombinant cTnI. Both mutations gave reduced inhibition of actin-tropomyosin-activated myosin ATPase, both in experiments using cTnI alone and in those using reconstituted human cardiac troponin under relaxing conditions. Both mutant troponin complexes also conferred increased calcium sensitivity of ATPase regulation. Studies on wild type/R145G mutant mixtures showed that the wild type phenotype was dominant in that the inhibition and the calcium sensitivity conferred by a 50:50 mixture was more similar to wild type than expected. Surface plasmon resonance-based assays showed that R162W mutant had an increased affinity for troponin C in the presence of calcium. This observation may contribute to the increased calcium sensitivity found with this mutant and also corroborates recent structural data. The observed decreased inhibition and increased calcium sensitivity suggest that these mutations may cause HCM via impaired relaxation rather than the impaired contraction seen with some other classes of HCM mutants. PMID- 10806206 TI - Evidence for the existence of a non-catalytic modifier site of peptide hydrolysis by the 20 S proteasome. AB - The 20 S proteasome is an endoprotease complex that preferentially cleaves peptides C-terminal of hydrophobic, basic, and acidic residues. Recently, we showed that these specific activities, classified as chymotrypsin-like, trypsin like, and peptidylglutamyl peptide-hydrolyzing (PGPH) activity, are differently affected by Ritonavir, an inhibitor of human immunodeficiency virus-1 protease. Ritonavir competitively inhibited the chymotrypsin-like activity, whereas the trypsin-like activity was enhanced. Here we demonstrate that the Ritonavir mediated up-regulation of the trypsin-like activity is not affected by specific active site inhibitors of the chymo-trypsin-like and PGPH activity. Moreover, we show that the mutual regulation of chymotrypsin-like and PGPH activities by their substrates as described previously by a "cyclical bite-chew" model is not affected by selective inhibitors of the respective active sites. These data challenge the bite-chew model and suggest that effectors of proteasome activity can act by binding to non-catalytic sites. Accordingly, we propose a kinetic "two site modifier" model that assumes that the substrate (or effector) may bind to an active site as well as to a second non-catalytic modifier site. This model appears to be valid as it describes the complex kinetic effects of Ritonavir very well. Since Ritonavir partially inhibits major histocompatibility complex class I restricted antigen presentation, the postulated modifier site may be required to coordinate the active centers of the proteasome for the production of class I peptide ligands. PMID- 10806207 TI - Control sites of ribosomal S6 kinase B and persistent activation through tumor necrosis factor. AB - RSKB, a 90-kDa ribosomal S6 protein kinase family (RSK) member with two complete catalytic domains connected by a linker, is activated through p38- and ERK mitogen-activated protein kinases. The N-terminal kinases of RSKs phosphorylate substrates; activation requires phosphorylation of linker and C-terminal kinase sites. Unlike other RSKs, the activation loop phosphorylation sites of both catalytic domains of RSKB, Ser(196) and Thr(568), were required for activity. RSKB activation depended on phosphorylation of linker Ser(343) and Ser(360) and associated with phosphorylation of nonconserved Ser(347), but Ser(347)-deficient RSKB retained partial activity. The known protein kinase A and protein kinase C inhibitors, H89 and Ro31-8220, blocked RSKB activity. Treatment of HeLa cells with tumor necrosis factor, epidermal growth factor, phorbol 12-myristate 13 acetate, and ionomycin but not with insulin resulted in strong activation of endogenous RSKB. High RSKB activity and Ser(347)/Ser(360) phosphorylation persisted for 3 h in tumor necrosis factor-treated cells, in contrast to the short bursts of p38, ERK, and RSK1-3 activities. In conclusion, a variety of stimuli induced phosphorylation and activation of RSKB through both p38 and ERK pathways; the persistence of activation indicated that RSKB selectively escaped cell mechanisms causing rapid deactivation of upstream p38 and ERK and other RSKs. PMID- 10806208 TI - Potentiation of endogenous fibrinolysis and rescue from lung ischemia/reperfusion injury in interleukin (IL)-10-reconstituted IL-10 null mice. AB - Little is known about interactions between endogenous anti-inflammatory paradigms and microvascular thrombosis in lung ischemia/reperfusion (I/R) injury. Interleukin (IL)-10 suppresses macrophage activation and down-regulates proinflammatory cytokine production, but there are no available data to suggest a link between IL-10, thrombosis, and fibrinolysis in the setting of I/R. We hypothesized that hypoxia/ischemia triggers IL-10 production, to dampen proinflammatory cytokine and adhesion receptor cascades and to restore vascular patency by fibrinolytic potentiation. Studies were performed in a mouse lung I/R model. IL-10 mRNA levels in lung were increased 43-fold over base line by 1 h of ischemia/2 h of reperfusion, with a corresponding increase in plasma IL-10. Expression was prominently localized in bronchial epithelial cells and mononuclear phagocytes. To study the link between IL-10 and fibrinolysis in vivo, the induction of plasminogen activator inhibitor-1 (PAI-1) was evaluated. Northern analysis demonstrated exaggerated pulmonary PAI-1 expression in IL-10 ( /-) mice after I/R, with a corresponding increase in plasma PAI/tissue-type plasminogen activator activity. In vivo, IL-10 (-/-) mice showed poor postischemic lung function and survival after I/R compared with IL-10 (+/+) mice. Despite a decrease in infiltration of mononuclear phagocytes in I/R lungs of IL 10 (-/-) mice, an increased intravascular pulmonary fibrin deposition was observed by immunohistochemistry and Western blotting, along with increased IL-1 expression. Recombinant IL-10 given to IL-10 (-/-) mice normalized the PAI/tissue type plasminogen activator ratio, reduced pulmonary vascular fibrin deposition, and rescued mice from lung injury. Since recombinant hirudin (direct thrombin inhibitor) also sufficed to rescue IL-10 (-/-) mice, these data suggest a preeminent role for microvascular thrombosis in I/R lung injury. Ischemia-driven IL-10 expression confers postischemic pulmonary protection by augmenting endogenous fibrinolytic mechanisms. PMID- 10806209 TI - Endotoxin-induced renal inflammatory response. Oncostatin M as a major mediator of suppressed renin expression. AB - The systemic response to endotoxin is characterized by hypotension and severe reductions in blood pressure, leading to cardiovascular collapse that can accompany septicemia. The renin/angiotensin system would normally be expected to respond to hypotensive challenge; however, inflammation appears to modify this response. This study identifies a strong acute phase response of the kidney that is characterized by enhanced expression of serum amyloid A, haptoglobin and tissue inhibitor for metalloproteinase-1 and a reduced expression of renin. Equivalent regulatory effects were observed for the immortalized As4.1 kidney cell line that models certain features of juxtaglomerular cells. Oncostatin M, a known endotoxin-responsive proinflammatory cytokine, proved to be an effective inhibitor of renin gene expression. Suppression by oncostatin M involves activated STAT5 and requires an inhibitory element in the renin promoter that functions separately from cell type-specific enhancer elements. The renal acute phase reaction, unlike the liver acute phase reaction, is more strongly dependent on locally produced inflammatory factors. PMID- 10806210 TI - Apurinic endonuclease activity of yeast Apn2 protein. AB - Abasic (apurinic/apyrimidinic; AP) sites are generated in vivo through spontaneous base loss and by enzymatic removal of bases damaged by alkylating agents and reactive oxygen species. In Saccharomyces cerevisiae, the APN1 and APN2 genes function in alternate pathways of AP site removal. Apn2-like proteins have been identified in other eukaryotes including humans, and these proteins form a distinct subfamily within the exonuclease III (ExoIII)/Ape1/Apn2 family of proteins. Apn2 and other members of this subfamily contain a carboxyl-terminal extension not present in the ExoIII/Ape1-like proteins. Here, we purify the Apn2 protein from yeast and show that it is a class II AP endonuclease. Deletion of the carboxyl terminus does not affect the AP endonuclease activity of the protein, but this protein is defective in the removal of AP sites in vivo. The carboxyl terminus may enable Apn2 to complex with other proteins, and such a multiprotein assembly may be necessary for the efficient recognition and cleavage of AP sites in vivo. PMID- 10806211 TI - Phosphorylation of the beta-amyloid precursor protein at the cell surface by ectocasein kinases 1 and 2. AB - The beta-amyloid precursor protein (betaAPP) is one of the rare proteins known to be phosphorylated within its ectodomain. We have shown previously that betaAPP can be phosphorylated within secretory vesicles and at the cell surface (Walter, J., Capell, A., Hung, A. Y. , Langen, H., Schnolzer, M., Thinakaran, G., Sisodia, S. S., Selkoe, D. J., and Haass, C. (1997) J. Biol. Chem. 272, 1896-1903). We have now specifically characterized the phosphorylation of cell surface-located betaAPP and identified two ectoprotein kinases that phosphorylate betaAPP at the outer face of the plasma membrane. By using selective protein kinase inhibitors and by investigating the usage of ATP and GTP as cosubstrates, we demonstrate that membrane-bound betaAPP as well as secreted forms of betaAPP can be phosphorylated by casein kinase (CK) 1- and CK2-like ectoprotein kinases. The ectodomain of betaAPP was also phosphorylated by purified CK1 and CK2 in vitro, but not by protein kinases A and C. Phosphorylation of betaAPP by ectoprotein kinases and by purified CK1 and CK2 occurred within an acidic domain in the N terminal half of the protein. Heparin strongly inhibited the phosphorylation of cell-surface betaAPP by ecto-CK1 and ecto-CK2, indicating a regulatory role of this extracellular matrix component in betaAPP phosphorylation. PMID- 10806212 TI - Phorbol 12-myristate 13-acetate induces protein kinase ceta-specific proliferative response in astrocytic tumor cells. AB - Protein kinase C (PKC) activation has been implicated in cellular proliferation in neoplastic astrocytes. The roles for specific PKC isozymes in regulating this glial response, however, are not well understood. The aim of this study was to characterize the expression of PKC isozymes and the role of PKC-eta expression in regulating cellular proliferation in two well characterized astrocytic tumor cell lines (U-1242 MG and U-251 MG) with different properties of growth in cell culture. Both cell lines expressed an array of conventional (alpha, betaI, betaII, and gamma) and novel (theta and epsilon) PKC isozymes that can be activated by phorbol myristate acetate (PMA). Another novel PKC isozyme, PKC-eta, was only expressed by U-251 MG cells. In contrast, PKC-delta was readily detected in U-1242 MG cells but was present only at low levels in U-251 MG cells. PMA (100 nm) treatment for 24 h increased cell proliferation by over 2-fold in the U-251 MG cells, whereas it decreased the mitogenic response in the U-1242 MG cells by over 90%. When PKC-eta was stably transfected into U-1242 MG cells, PMA increased cell proliferation by 2.2-fold, similar to the response of U-251 MG cells. The cell proliferation induced by PMA in both the U-251 MG and U-1242-PKC-eta cells was blocked by the PKC inhibitor bisindolylmaleimide (0.5 micrometer) and the MEK inhibitor, PD 98059 (50 micrometer). Transient transfection of wild type U-251 with PKC-eta antisense oligonucleotide (1 micrometer) also blocked the PMA induced increase in [(3)H]thymidine incorporation. The data demonstrate that two glioblastoma lines, with functionally distinct proliferative responses to PMA, express different novel PKC isozymes and that the differential expression of PKC eta plays a determining role in the different proliferative capacity. PMID- 10806213 TI - Structural analysis of glycosaminoglycans in animals bearing mutations in sugarless, sulfateless, and tout-velu. Drosophila homologues of vertebrate genes encoding glycosaminoglycan biosynthetic enzymes. AB - Mutations that disrupt developmental patterning in Drosophila have provided considerable information about growth factor signaling mechanisms. Three genes recently demonstrated to affect signaling by members of the Wnt, transforming growth factor-beta, Hedgehog, and fibroblast growth factor families in Drosophila encode proteins with homology to vertebrate enzymes involved in glycosaminoglycan synthesis. We report here the biochemical characterization of glycosaminoglycans in Drosophila bearing mutations in sugarless, sulfateless, and tout-velu. We find that mutations in sugarless, which encodes a protein with homology to UDP-glucose dehydrogenase, compromise the synthesis of both chondroitin and heparan sulfate, as would be predicted from a defect in UDP-glucuronate production. Defects in sulfateless, a gene encoding a protein with similarity to vertebrate N deacetylase/N-sulfotransferases, do not affect chondroitin sulfate levels or composition but dramatically alter the composition of heparin lyase-released disaccharides. N-, 6-O-, and 2-O-sulfated disaccharides are absent and replaced entirely with an unsulfated disaccharide. A mutation in tout-velu, a gene related to the vertebrate Exostoses 1 heparan sulfate co-polymerase, likewise does not affect chondroitin sulfate synthesis but reduces all forms of heparan sulfate to below the limit of detection. These findings show that sugarless, sulfateless, and tout-velu affect glycosaminoglycan biosynthesis and demonstrate the utility of Drosophila as a model organism for studying the function and biosynthesis of glycosaminoglycans in vivo. PMID- 10806214 TI - Heat shock protein 70 inhibits apoptosis downstream of cytochrome c release and upstream of caspase-3 activation. AB - Heat shock protein 70 (HSP70) has been shown to act as an inhibitor of apoptosis. We have also observed an inhibitory effect of HSP70 on apoptotic cell death both in preheated U937 and stably transfected HSP70-overexpressing U937 (U937/HSP70) cells. However, the molecular mechanism whereby HSP70 prevents apoptosis still remains to be solved. To address this issue, we investigated the effect of HSP70 on apoptotic processes in an in vitro system. Caspase-3 cleavage and DNA fragmentation were detected in cytosolic fractions from normal cells upon addition of dATP, but not from preheated U937 or U937/hsp70 cells. Moreover, the addition of purified recombinant HSP70 to normal cytosolic fractions prevented caspase-3 cleavage and DNA fragmentation, suggesting that HSP70 prevents apoptosis upstream of caspase-3 processing. Because cytochrome c was still released from mitochondria into the cytosol by lethal heat shock despite prevention of caspase-3 activation and cell death in both preheated U937 and U937/hsp70 cells, it was evident that HSP70 acts downstream of cytochrome c release. Results obtained in vitro with purified deletion mutants of HSP70 showed that the carboxyl one-third region (from amino acids 438 to 641) including the peptide-binding domain and the carboxyl-terminal EEVD sequence was essential to prevent caspase-3 processing. From these results, we conclude that HSP70 acts as a strong suppressor of apoptosis acting downstream of cytochrome c release and upstream of caspase-3 activation. PMID- 10806215 TI - Endogenous protein kinase CK2 participates in Wnt signaling in mammary epithelial cells. AB - Protein kinase CK2 (formerly casein kinase II) is a serine/threonine kinase overexpressed in many human tumors, transformed cell lines, and rapidly proliferating tissues. Recent data have shown that many cancers involve inappropriate reactivation of Wnt signaling through ectopic expression of Wnts themselves, as has been seen in a number of human breast cancers, or through mutation of intermediates in the Wnt pathway, such as adenomatous polyposis coli or beta-catenin, as described in colon and other cancers. Wnts are secreted factors that are important in embryonic development, but overexpression of certain Wnts, such as Wnt-1, leads to proliferation and transformation of cells. We report that upon stable transfection of Wnt-1 into the mouse mammary epithelial cell line C57MG, morphological changes and increased proliferation are accompanied by increased levels of CK2, as well as of beta-catenin. CK2 and beta catenin co-precipitate with the Dvl proteins, which are Wnt signaling intermediates. A major phosphoprotein of the size of beta-catenin appears in in vitro kinase reactions performed on the Dvl immunoprecipitates. In vitro translated beta-catenin, Dvl-2, and Dvl-3 are phosphorylated by CK2. The selective CK2 inhibitor apigenin blocks proliferation of Wnt-1-transfected cells, abrogates phosphorylation of beta-catenin, and reduces beta-catenin and Dvl protein levels. These results demonstrate that endogenous CK2 is a positive regulator of Wnt signaling and growth of mammary epithelial cells. PMID- 10806216 TI - Coordination between the polymerase and 5'-nuclease components of DNA polymerase I of Escherichia coli. AB - The polymerase and 5'-nuclease components of DNA polymerase I must collaborate in vivo so as to generate ligatable structures. Footprinting shows that the polymerase and 5'-nuclease cannot bind simultaneously to a DNA substrate and appear to compete with one another, suggesting that the two active sites are physically separate and operate independently. The desired biological end point, a ligatable nick, results from the substrate specificities of the polymerase and 5'-nuclease. The preferred substrate of the 5'-nuclease is a "double-flap" structure having a frayed base at the primer terminus overlapping the displaced strand that is to be cleaved by the 5'-nuclease. Cleavage of this structure occurs almost exclusively between the first two paired bases of the downstream strand, yielding a ligatable nick. In whole DNA polymerase I, the polymerase and 5'-nuclease activities are coupled such that the majority of molecules cleaved by the 5'-nuclease have also undergone polymerase-catalyzed addition to the primer terminus. This implies that the 5'-nuclease can capture a DNA molecule from the polymerase site more efficiently than from the bulk solution. PMID- 10806217 TI - Impaired estrogen sensitivity in bone by inhibiting both estrogen receptor alpha and beta pathways. AB - Although it is well established that estrogen deficiency causes osteoporosis among the postmenopausal women, the involvement of estrogen receptor (ER) in its pathogenesis still remains uncertain. In the present study, we have generated rats harboring a dominant negative ERalpha, which inhibits the actions of not only ERalpha but also recently identified ERbeta. Contrary to our expectation, the bone mineral density (BMD) of the resulting transgenic female rats was maintained at the same level with that of the wild-type littermates when sham operated. In addition, ovariectomy-induced bone loss was observed almost equally in both groups. Strikingly, however, the BMD of the transgenic female rats, after ovariectomized, remained decreased even if 17beta-estradiol (E(2)) was administrated, whereas, in contrast, the decrease of littermate BMD was completely prevented by E(2). Moreover, bone histomorphometrical analysis of ovariectomized transgenic rats revealed that the higher rates of bone turnover still remained after treatment with E(2). These results demonstrate that the prevention from the ovariectomy-induced bone loss by estrogen is mediated by ER pathways and that the maintenance of BMD before ovariectomy might be compensated by other mechanisms distinct from ERalpha and ERbeta pathways. PMID- 10806219 TI - Broadening the scope while maintaining high impact PMID- 10806220 TI - Optical coherence microscopy. A technology for rapid, in vivo, non-destructive visualization of plants and plant cells. AB - We describe the development and utilization of a new imaging technology for plant biology, optical coherence microscopy (OCM), which allows true in vivo visualization of plants and plant cells. This novel technology allows the direct, in situ (e.g. plants in soil), three-dimensional visualization of cells and events in shoot tissues without causing damage. With OCM we can image cells or groups of cells that are up to 1 mm deep in living tissues, resolving structures less than 5 microm in size, with a typical collection time of 5 to 6 min. OCM measures the inherent light-scattering properties of biological tissues and cells. These optical properties vary and provide endogenous developmental markers. Singly scattered photons from small (e.g. 5 x 5 x 10 microm) volume elements (voxels) are collected, assembled, and quantitatively false-colored to form a three-dimensional image. These images can be cropped or sliced in any plane. Adjusting the colors and opacities assigned to voxels allows us to enhance different features within the tissues and cells. We show that light-scattering properties are the greatest in regions of the Arabidopsis shoot undergoing developmental processes. In large cells, high light scattering is produced from nuclei, intermediate light scatter is produced from cytoplasm, and little if any light scattering originates from the vacuole and cell wall. OCM allows the rapid, repetitive, non-destructive collection of quantitative data about inherent properties of cells, so it provides a means of continuously monitoring plants and plant cells during development and in response to exogenous stimuli. PMID- 10806218 TI - ERKs and p38 kinases mediate ultraviolet B-induced phosphorylation of histone H3 at serine 10. AB - Histone H3 is the core protein of the nucleosome. Phosphorylation of H3 involves immediate early gene expression, chromatin remodeling, and chromosome condensation during mitosis. Very recently, Rsk2 or MSK1 kinase-mediated phosphorylation of H3 at serine 10 was reported. In the present study, we show that both ERKs and p38 kinase may mediate ultraviolet B-induced phosphorylation of H3 at serine 10. PD 98059, a MEK1 inhibitor, and SB 202190, a p38 kinase inhibitor, efficiently inhibited ultraviolet B-induced phosphorylation of H3. Phosphorylation of H3 was also inhibited in cells expressing dominant negative mutant (DNM) ERK2 and DNM p38 kinase. In contrast, no inhibition of H3 phosphorylation in Jnk1 or Jnk2 knockout cells (Jnk1(-/-) or Jnk2(-/-)) and cells expressing DNM JNK1 was observed. More importantly, incubation of active ERK2 or p38 kinase with H3 protein resulted in phosphorylation of H3 at serine 10 in vitro. These results suggest that ERK and p38 kinase are at least two important mediators of phosphorylation of H3 at serine 10. PMID- 10806221 TI - Crop biotechnology. Where now? AB - Nature Biotechnology organized a conference in London on Agobiotech 99: Biotechnology and World Agriculture (November 14-16, 1999). The conference focused entirely on crop biotechnology and covered both societal and scientific aspects. Below is an account of the more important issues raised by the speakers and the audience. PMID- 10806222 TI - Solar-powered sea slugs. Mollusc/algal chloroplast symbiosis. PMID- 10806223 TI - Photoreceptors and regulation of flowering time. PMID- 10806224 TI - Molecular cloning and targeting of a fibrillarin homolog from Arabidopsis. AB - Fibrillarin is a nucleolar protein known to be involved in the processing of ribosomal RNA precursors. We isolated AtFbr1, a cDNA encoding a homolog of fibrillarin in Arabidopsis. The cDNA is 1.2 kb in size and encodes a polypeptide of 310 amino acid residues with a molecular mass of 33 kD. AtFbr1 is expressed at high levels in the flower and root tissue and at a slightly lower level in leaf tissue, whereas it was nearly undetectable in siliques. Expression of AtFbr1 was compared with that of the FLP (fibrillarin-like protein) gene identified by the Arabidopsis genome project. Abscisic acid treatment resulted in the down regulation of the expression of both AtFbr1 and FLP genes in seedlings, although the degree of suppression was higher for FLP than for AtFbr1. In addition, the expression level of FLP decreased with the age of the seedlings, whereas AtFbr1 did not exhibit any detectable change. The subcellular localization of AtFbrl was studied with an in vivo targeting approach using a fusion protein, and was found to be correctly targeted to the nucleolus in protoplasts when expressed as a green fluorescent fusion protein (GFP). Deletion experiments showed that the N terminal glycine- and arginine-rich region is necessary and sufficient to target AtFbr1 to the nucleolus. PMID- 10806225 TI - Ectopic deposition of lignin in the pith of stems of two Arabidopsis mutants. AB - The biosynthesis of lignin in vascular plants is regulated both developmentally and environmentally. In the inflorescence stems of Arabidopsis, lignin is mainly deposited in the walls of xylem cells and interfascicular fiber cells during normal plant growth and development. The mechanisms controlling the spatial deposition of lignin remain unknown. By screening ethyl methanesulfonate mutagenized populations of Arabidopsis, we have isolated two allelic elp1 (ectopic deposition of lignin in pith) mutants with altered lignin deposition patterns. In elp1 stems, lignin was ectopically deposited in the walls of pith parenchyma cells in addition to its normal deposition in the walls of xylem and fiber cells. Lignin appeared to be deposited in patches of parenchyma cells in the pith of both young and mature elp1 stems. The ectopic deposition of lignin in the pith of elp1 stems was accompanied by an increase in the activities of enzymes in the lignin biosynthetic pathway and with the ectopic expression of caffeoyl coenzyme A O-methyltransferase in pith cells. These results indicate that the ELP1 locus is involved in the repression of the lignin biosynthetic pathway in the pith. Isolation of the elp1 mutants provides a novel means with which to study the molecular mechanisms underlying the spatial control of lignification. PMID- 10806226 TI - Cloning, developmental, and tissue-specific expression of sucrose:sucrose 1 fructosyl transferase from Taraxacum officinale. Fructan localization in roots. AB - Sucrose:sucrose 1-fructosyl transferase (1-SST) is the key enzyme initiating fructan synthesis in Asteraceae. Using reverse transcriptase-PCR, we isolated the cDNA for 1-SST from Taraxacum officinale. The cDNA-derived amino acid sequence showed very high homology to other Asteracean 1-SSTs (Cichorium intybus 86%, Cynara scolymus 82%, Helianthus tuberosus 80%), but homology to 1-SST from Allium cepa (46%) and Aspergillus foetidus (18%) was much lower. Fructan concentrations, 1-SST activities, 1-SST protein, and mRNA concentrations were compared in different organs during vegetative and generative development of T. officinale plants. Expression of 1-SST was abundant in young roots but very low in leaves. 1 SST was also expressed at the flowering stages in roots, stalks, and receptacles. A good correlation was found between northern and western blots showing transcriptional regulation of 1-SST. At the pre-flowering stage, 1-SST mRNA concentrations and 1-SST activities were higher in the root phloem than in the xylem, resulting in the higher fructan concentrations in the phloem. Fructan localization studies indicated that fructan is preferentially stored in phloem parenchyma cells in the vicinity of the secondary sieve tube elements. However, inulin-like crystals occasionally appeared in xylem vessels. PMID- 10806227 TI - Response to Xanthomonas campestris pv. vesicatoria in tomato involves regulation of ethylene receptor gene expression. AB - Although ethylene regulates a wide range of defense-related genes, its role in plant defense varies greatly among different plant-microbe interactions. We compared ethylene's role in plant response to virulent and avirulent strains of Xanthomonas campestris pv. vesicatoria in tomato (Lycopersicon esculentum Mill.). The ethylene-insensitive Never ripe (Nr) mutant displays increased tolerance to the virulent strain, while maintaining resistance to the avirulent strain. Expression of the ethylene receptor genes NR and LeETR4 was induced by infection with both virulent and avirulent strains; however, the induction of LeETR4 expression by the avirulent strain was blocked in the Nr mutant. To determine whether ethylene receptor levels affect symptom development, transgenic plants overexpressing a wild-type NR cDNA were infected with virulent X. campestris pv. vesicatoria. Like the Nr mutant, the NR overexpressors displayed greatly reduced necrosis in response to this pathogen. NR overexpression also reduced ethylene sensitivity in seedlings and mature plants, indicating that, like LeETR4, this receptor is a negative regulator of ethylene response. Therefore, pathogen induced increases in ethylene receptors may limit the spread of necrosis by reducing ethylene sensitivity. PMID- 10806228 TI - Characterization of brassinazole, a triazole-type brassinosteroid biosynthesis inhibitor. AB - Screening for brassinosteroid (BR) biosynthesis inhibitors was performed to find chemicals that induce dwarfism in Arabidopsis, mutants that resembled BR biosynthesis mutants that can be rescued by BR. Through this screening experiment, the compound brassinazole was selected as the most potent chemical. In dark-grown Arabidopsis, brassinazole-induced morphological changes were nearly restored to those of wild type by treatment with brassinolide. The structure of brassinazole is similar to pacrobutrazol, a gibberellin biosynthesis inhibitor. However, in assays with cress (Lepidium sativum) plants, brassinazole-treated plants did not show recovery after the addition of gibberellin but showed good recovery after the addition of brassinolide. These data demonstrate that brassinazole is a specific BR biosynthesis inhibitor. Brassinazole-treated cress also showed dwarfism, with altered leaf morphology, including the downward curling and dark green color typical of Arabidopsis BR-deficient mutants, and this dwarfism was reversed by the application of 10 nM brassinolide. This result suggests that BRs are essential for plant growth, and that brassinazole can be used to clarify the function of BRs in plants as a complement to BR-deficient mutants. The brassinazole action site was also investigated by feeding BR biosynthesis intermediates to cress grown in the light. PMID- 10806229 TI - Mutation in the threonine synthase gene results in an over-accumulation of soluble methionine in Arabidopsis. AB - In higher plants, O-phosphohomoserine (OPH) represents a branch point between the methionine (Met) and threonine (Thr) biosynthetic pathways. It is believed that the enzymes Thr synthase (TS) and cystathionine gamma-synthase (CGS) actively compete for the OPH substrate for Thr and Met biosynthesis, respectively. We have isolated a mutant of Arabidopsis, designated mto2-1, that over-accumulates soluble Met 22-fold and contains markedly reduced levels of soluble Thr in young rosettes. The mto2-1 mutant carries a single base pair mutation within the gene encoding TS, resulting in a leucine-204 to arginine change. Accumulation of TS mRNA and protein was normal in young rosettes of mto2-1, whereas functional complementation analysis of an Escherichia coli thrC mutation suggested that the ability of mto2-1 TS to synthesize Thr is impaired. We concluded that the mutation within the TS gene is responsible for the mto2-1 phenotype, resulting in decreased Thr biosynthesis and a channeling of OPH to Met biosynthesis in young rosettes. Analysis of the mto2-1 mutant suggested that, in vivo, the feedback regulation of CGS is not sufficient alone for the control of Met biosynthesis in young rosettes and is dependent on TS activity. In addition, developmental analysis of soluble Met and Thr concentrations indicated that the accumulation of these amino acids is regulated in a temporal and spatial manner. PMID- 10806230 TI - Expression of water channel proteins in Mesembryanthemum crystallinum. AB - We have characterized transcripts for nine major intrinsic proteins (MIPs), some of which function as water channels (aquaporins), from the ice plant Mesembryanthemum crystallinum. To determine the cellular distribution and expression of these MIPs, oligopeptide-based antibodies were generated against MIP-A, MIP-B, MIP-C, or MIP-F, which, according to sequence and functional characteristics, are located in the plasma membrane (PM) and tonoplast, respectively. MIPs were most abundant in cells involved in bulk water flow and solute flux. The tonoplast MIP-F was found in all cells, while signature cell types identified different PM-MIPs: MIP-A predominantly in phloem-associated cells, MIP-B in xylem parenchyma, and MIP-C in the epidermis and endodermis of immature roots. Membrane protein analysis confirmed MIP-F as tonoplast located. MIP-A and MIP-B were found in tonoplast fractions and also in fractions distinct from either the tonoplast or PM. MIP-C was most abundant but not exclusive to PM fractions, where it is expected based on its sequence signature. We suggest that within the cell, MIPs are mobile, which is similar to aquaporins cycling through animal endosomes. MIP cycling and the differential regulation of these proteins observed under conditions of salt stress may be fundamental for the control of tissue water flux. PMID- 10806231 TI - Sieve tube unloading and post-phloem transport of fluorescent tracers and proteins injected into sieve tubes via severed aphid stylets. AB - A variety of fluorescent tracers and proteins were injected via severed aphid stylets into the sieve tubes of wheat (Triticum aestivum L.) grains to evaluate the dimensions of plasmodesmal channels involved in sieve element/companion cell (SE/CC) unloading and post-phloem transport. In the post-phloem pathway, where diffusion is the predominant mode of transport, the largest molecule to show mobility was 16-kD dextran, with a Stokes radius of 2.6 nm. This suggests that the aqueous channels for cell-to-cell transport must be about 8 nm in diameter. Even the largest tracer injected into the sieve tubes, 400-kD fluorescein-labeled Ficoll with a Stokes radius of about 11 nm, was unloaded from the SE/CC complex. However, in contrast to smaller tracers (< or =3 kD, with a Stokes radius < or = 1.2 nm), the unloading of fluorescein-labeled Ficoll and other large molecules from the SE/CC complex showed an irregular, patchy distribution, with no further movement along the post-phloem pathway. Either the plasmodesmal channels involved in SE/CC unloading are exceptionally large (perhaps as much as 42 nm in diameter), with only a very small fraction of plasmodesmata being conductive, or the larger tracers damage the plasmodesmata in some way, enlarging smaller channels. PMID- 10806232 TI - Gradients in water potential and turgor pressure along the translocation pathway during grain filling in normally watered and water-stressed wheat plants. AB - The water relations parameters involved in assimilate flow into developing wheat (Triticum aestivum L.) grains were measured at several points from the flag leaf to the endosperm cavity in normally watered (Psi approximately -0.3 MPa) and water-stressed plants (Psi approximately -2 MPa). These included direct measurement of sieve tube turgor and several independent approaches to the measurement or calculation of water potentials in the peduncle, grain pericarp, and endosperm cavity. Sieve tube turgor measurements, osmotic concentrations, and Psi measurements using dextran microdrops showed good internal consistency (i.e. Psi = Psi(s) + Psi(p)) from 0 to -4 MPa. In normally watered plants, crease pericarp Psi and sieve tube turgor were almost 1 MPa lower than in the peduncle. This suggests a high hydraulic resistance in the sieve tubes connecting the two. However, observations concerning exudation rates indicated a low resistance. In water-stressed plants, peduncle Psi and crease pericarp Psi were similar. In both treatments, there was a variable, approximately 1-MPa drop in turgor pressure between the grain sieve tubes and vascular parenchyma cells. There was little between-treatment difference in endosperm cavity sucrose or osmotic concentrations or in the crease pericarp sucrose pool size. Our results re emphasize the importance of the sieve tube unloading step in the control of assimilate import. PMID- 10806233 TI - Physiological implications of the kinetics of maize leaf phosphoenolpyruvate carboxylase. AB - It has been a common practice to assay phosphoenolpyruvate carboxylase (PEPC) under high, nonphysiological concentrations of Mg(2+) and bicarbonate. We have performed kinetic studies on the enzyme from maize (Zea mays) leaves at near physiological levels of free Mg(2+) (0.4 mM) and bicarbonate (0.1 mM), and found that both the nonphosphorylated and phosphorylated enzymes exhibited a high degree of cooperativity in the binding of phosphoenolpyruvate, a much lower affinity for this substrate and for activators, and a greater affinity for malate than at high concentrations of these ions. Inhibition of the phosphorylated enzyme by malate was overcome by glycine or alanine but not by glucose-6 phosphate, either in the absence or presence of high concentrations of glycerol, a compatible solute. Alanine caused significant activation at physiological concentrations, suggesting a pivotal role for this amino acid in regulating maize leaf PEPC activity. Our results showed that the maximum enzyme activity attainable in vivo would be less than 50% of that attainable in vitro under optimum conditions. Therefore, the high levels of PEPC protein in the cytosol of C(4) mesophyll cells might be an adaptation for sustaining the steady-state rate of flux through the photosynthetic CO(2) assimilation pathway despite the limitations imposed by the PEPC kinetic properties and the conditions of its environment. PMID- 10806234 TI - Use of recombinant aequorin to study calcium homeostasis and monitor calcium transients in response to heat and cold shock in cyanobacteria. AB - We investigated the possibility of Ca(2+) signaling in cyanobacteria (blue-green algae) by measuring intracellular free Ca(2+) levels ([Ca(2+)](i)) in a recombinant strain of the nitrogen fixing cyanobacterium Anabaena strain sp. PCC7120, which constitutively expresses the Ca(2+)-binding photoprotein apoaequorin. The homeostasis of intracellular Ca(2+) in response to increasing external Ca(2+) has been studied in this strain. The resting level of free Ca(2+) in Anabaena was found to be between 100 and 200 nM. Additions of increasing concentrations of external Ca(2+) gave a transient burst of [Ca(2+)](i) followed by a very quick decline, reaching a plateau within seconds that brought the level of [Ca(2+)](i) back to the resting value. These results indicate that Anabaena strain sp. PCC7120 is able to regulate its internal Ca(2+) levels. We also monitored Ca(2+) transients in our recombinant strain in response to heat and cold shock. The cell's response to both stresses was dependent on the way they were induced. The use of inhibitors suggests that heat shock mobilizes cytosolic Ca(2+) from both intracellular and extracellular sources, while the Ca(2+) source for cold shock signaling is mostly extracellular. PMID- 10806235 TI - Octadecanoid-derived alteration of gene expression and the "oxylipin signature" in stressed barley leaves. Implications for different signaling pathways. AB - Stress-induced gene expression in barley (Hordeum vulgare cv Salome) leaves has been correlated with temporally changing levels of octadecanoids and jasmonates, quantified by means of gas chromatography/mass spectrometry-single ion monitoring. Application of sorbitol-induced stress led to a low and transient rise of jasmonic acid (JA), its precursor 12-oxophytodienoic acid (OPDA), and the methyl esters JAME and OPDAME, respectively, followed by a large increase in their levels. JA and JAME peaked between 12 and 16 h, about 4 h before OPDA and OPDAME. However, OPDA accumulated up to a 2.5-fold higher level than the other compounds. Dihomo-JA and 9, 13-didehydro-OPDA were identified as minor components. Kinetic analyses revealed that a transient threshold of jasmonates or octadecanoids is necessary and sufficient to initiate JA-responsive gene expression. Although OPDA and OPDAME applied exogenously were metabolized to JA in considerable amounts, both of them can induce gene expression, as evidenced by those genes that did not respond to endogenously formed JA. Also, coronatine induces JA-responsive genes independently from endogenous JA. Application of deuterated JA showed that endogenous synthesis of JA is not induced by JA treatment. The data are discussed in terms of distinct signaling pathways. PMID- 10806236 TI - The determination time of the carpel whorl is differentially sensitive to carbohydrate supply in Pharbitis nil. AB - A shoot apical meristem is florally determined if, following its removal from an induced plant, it flowers when cultured in non-inductive conditions. Determination times were measured in the short-day plant Pharbitis nil to examine whether floral whorls are determined simultaneously or sequentially. Shoot apices were excised at daily intervals following a 48-h dark-inductive treatment, cultured in non-inductive conditions for 4 weeks in continuous light, and the number of floral organs scored. The culture medium was White's supplemented with sucrose, glucose (Glc), fructose (Fru), or 1:1 Glc:Fru at 2% (w/v), 4% (w/v), or 6% (w/v) or sugar-mannitol combinations of osmotic potentials equivalent to 4% (w/v) or 6% (w/v). The minimum whorl determination time was 1 d for sepals, petals, and stamens regardless of carbon supply. However, for carpels it varied remarkably from 5 d on sucrose, to 2 to 3 d on Fru or Glc:Fru, to 1 d for 2% (w/v) and 6% (w/v) Glc. Therefore, depending on the carbon supply, the carpel whorl was determined at the same time or after the outer whorls. Generally, these effects could not be reproduced on the sugar-mannitol treatments. PMID- 10806237 TI - Internal conductance to CO(2) diffusion and C(18)OO discrimination in C(3) leaves. AB - (18)O discrimination in CO(2) stems from the oxygen exchange between (18)O enriched water and CO(2) in the chloroplast, a process catalyzed by carbonic anhydrase (CA). A proportion of this (18)O-labeled CO(2) escapes back to the atmosphere, resulting in an effective discrimination against C(18)OO during photosynthesis (Delta(18)O). By constraining the delta(18)O of chloroplast water (delta(e)) by analysis of transpired water and the extent of CO(2)-H(2)O isotopic equilibrium (theta(eq)) by measurements of CA activity (theta(eq) = 0.75-1.0 for tobacco, soybean, and oak), we could apply measured Delta(18)O in a leaf cuvette attached to a mass spectrometer to derive the CO(2) concentration at the physical limit of CA activity, i.e. the chloroplast surface (c(cs)). From the CO(2) drawdown sequence between stomatal cavities from gas exchange (c(i)), from Delta(18)O (c(cs)), and at Rubisco sites from Delta(13)C (c(c)), the internal CO(2) conductance (g(i)) was partitioned into cell wall (g(w)) and chloroplast (g(ch)) components. The results indicated that g(ch) is variable (0.42-1.13 mol m(-2) s(-1)) and proportional to CA activity. We suggest that the influence of CA activity on the CO(2) assimilation rate should be important mainly in plants with low internal conductances. PMID- 10806238 TI - The BtpA protein stabilizes the reaction center proteins of photosystem I in the cyanobacterium Synechocystis sp. PCC 6803 at low temperature. AB - Specific inhibition of photosystem I (PSI) was observed under low-temperature conditions in the cyanobacterium Synechocystis sp. strain PCC 6803. Growth at 20 degrees C caused inhibition of PSI activity and increased degradation of the PSI reaction center proteins PsaA and PsaB, while no significant changes were found in the level and activity of photosystem II (PSII). BtpA, a recently identified extrinsic thylakoid membrane protein, was found to be a necessary regulatory factor for stabilization of the PsaA and PsaB proteins under such low-temperature conditions. At normal growth temperature (30 degrees C), the BtpA protein was present in the cell, and its genetic deletion caused an increase in the degradation of the PSI reaction center proteins. However, growth of Synechocystis cells at 20 degrees C or shifting of cultures grown at 30 degrees C to 20 degrees C led to a rapid accumulation of the BtpA protein, presumably to stabilize the PSI complex, by lowering the rates of degradation of the PsaA and PsaB proteins. A btpA deletion mutant strain could not grow photoautotrophically at low temperature, and exhibited rapid degradation of the PSI complex after transfer of the cells from normal to low temperature. PMID- 10806239 TI - Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. AB - We studied the response of each ascorbate peroxidase (APX) isoenzyme in spinach leaves under stress conditions imposed by high light intensity, drought, salinity, and applications of methyl viologen and abscisic acid. The steady-state transcript level of cytosolic APX remarkably increased in response to high-light stress and methyl viologen treatment, but not in response to the other stress treatments. The transcript levels of the chloroplastic (stromal and thylakoid bound) and microbody-bound APX isoenzymes were not changed in response to any of the stress treatments. To explore the responses of the APX isoenzymes to photooxidative stress, the levels of transcript and protein and activities of each isoenzyme were studied during high-light stress and following its recovery. The cytosolic APX activity increased in parallel with transcript abundance during high-light stress, while the protein level was not altered. The other isoenzymes showed no significant changes in transcript and protein levels and activities, except for the gradual decrease in chloroplastic isoenzyme activities. PMID- 10806240 TI - fhy3-1 retains inductive responses of phytochrome A. AB - The fhy3 mutation of Arabidopsis impairs phytochrome A (phyA)-mediated inhibition of hypocotyl growth without affecting the levels of phyA measured spectrophotometrically or immunochemically. We investigated whether the fhy3-1 mutation has similar effects on very low fluence responses (VLFR) and high irradiance responses (HIR) of phyA. When exposed to hourly pulses of far-red light, etiolated seedlings of the wild type or of the fhy3-1 mutant showed similar inhibition of hypocotyl growth, unfolding of the cotyledons, anthocyanin synthesis, and greening upon transfer to white light. In the wild type, continuous far-red light was significantly more effective than hourly far-red pulses (at equal total fluence). In the fhy3-1 mutant, hourly pulses were as effective as continuous far-red light, i.e. the failure of reciprocity typical of HIR was not observed. Germination was similarly promoted by continuous or pulsed far-red in wild-type and fhy3-1 seeds. Thus, for hypocotyl growth, cotyledon unfolding, greening, and seed germination, the fhy3-1 mutant retains VLFR but is severely impaired in HIR. These data are consistent with the idea that VLFR and HIR involve divergent signaling pathways of phyA. PMID- 10806241 TI - Comparative analysis of expressed sequences in Phytophthora sojae. AB - Phytophthora sojae (Kaufmann and Gerdemann) is an oomycete that causes stem and root rot on soybean (Glycine max L. Merr) plants. We have constructed three cDNA libraries using mRNA isolated from axenically grown mycelium and zoospores and from tissue isolated from plant hypocotyls 48 h after inoculation with zoospores. A total of 3,035 expressed sequence tags (ESTs) were generated from the three cDNA libraries, representing an estimated 2,189 cDNA transcripts. The ESTs were classified according to putative function based on similarity to known proteins, and were analyzed for redundancy within and among the three source libraries. Distinct expression patterns were observed for each library. By analysis of the percentage G+C content of the ESTs, we estimate that two-thirds of the ESTs from the infected plant library are derived from P. sojae cDNA transcripts. The ESTs originating from this study were also compared with a collection of Phytophthora infestans ESTs and with all other non-human ESTs to assess the similarity of the P. sojae sequences to existing EST data. This collection of cDNA libraries, ESTs, and accompanying annotation will provide a new resource for studies on oomycetes and on soybean responses to pathogen challenge. PMID- 10806242 TI - Differential regulation of plastidial and cytosolic isoforms of peptide methionine sulfoxide reductase in Arabidopsis. AB - We report the characterization of two members of a gene family from Arabidopsis that encode, respectively, cytosolic (cPMSR) and plastid-targeted (pPMSR) isoforms of the oxidative-stress-repair enzyme peptide methionine sulfoxide reductase. Overexpression of these proteins in Escherichia coli confirmed that each had PMSR enzyme activity with a synthetic substrate, N-acetyl-[(3)H] methionine sulfoxide, or a biological substrate, alpha-1 proteinase inhibitor. The pPMSR was imported into intact chloroplasts in vitro with concomitant cleavage of its approximately 5-kD N-terminal signal peptide. The two PMSR isoforms exhibited divergent pH optima, tissue localization, and responses to developmental and environmental effects. Analysis of the Arabidopsis database indicated that there are probably at least two p-pmsr-like genes and three c-pmsr like genes in the Arabidopsis genome. Expression of the p-pmsr genes and their protein products was restricted to photosynthetic tissues and was strongly induced following illumination of etiolated seedlings. In contrast, the c-pmsr genes were expressed at moderate levels in all tissues and were only weakly affected by light. Exposure to a variety of biotic and abiotic stresses showed relatively little effect on pmsr gene expression, with the exception of leaves subjected to a long-term exposure to the cauliflower mosaic virus. These leaves showed a strong induction of the c-pmsr gene after 2 to 3 weeks of chronic pathogen infection. These data suggest novel roles for PMSR in photosynthetic tissues and in pathogen defense responses in plants. PMID- 10806243 TI - Disaccharide-mediated regulation of sucrose:fructan-6-fructosyltransferase, a key enzyme of fructan synthesis in barley leaves. AB - Previous work has indicated that sugar sensing may be important in the regulation of fructan biosynthesis in grasses. We used primary leaves of barley (Hordeum vulgare cv Baraka) to study the mechanisms involved. Excised leaf blades were supplied in the dark with various carbohydrates. Fructan pool sizes and two key enzymes of fructan biosynthesis, sucrose (Suc):Suc-1-fructosyltransferase (1-SST; EC 2. 4.1.99) and Suc:fructan-6-fructosyltransferase (6-SFT; EC 2.4.1.10) were analyzed. Upon supply of Suc, fructan pool sizes increased markedly. Within 24 h, 1-SST activity was stimulated by a factor of three and 6-SFT-activity by a factor of more than 20, compared with control leaves supplemented with mannitol (Mit). At the same time, the level of mRNA encoding 6-SFT increased conspicuously. These effects were increased in the presence of the invertase inhibitor 2, 5-dideoxy 2,5-imino-D-mannitol. Compared with equimolar solutions of Suc, glucose (Glu) and fructose stimulated 6-SFT activity to a lesser extent. Remarkably, trehalose (Tre; Glc-alpha-1 and 1-alpha-Glc) had stimulatory effects on 6-SFT activity and, to a somewhat lesser extent, on 6-SFT mRNA, even in the presence of validoxylamine A, a potent trehalase inhibitor. Tre by itself, however, in the presence or absence of validoxylamine A, did not stimulate fructan accumulation. Monosaccharides phosphorylated by hexokinase but not or weakly metabolized, such as mannose (Man) or 2-deoxy-Glc, had no stimulatory effects on fructan synthesis. When fructose or Man were supplied together with Tre, fructan and starch biosynthesis were strongly stimulated. Concomitantly, phospho-Man isomerase (EC 5.3.1.8) activity was detected. These results indicate that the regulation of fructan synthesis in barley leaves occurs independently of hexokinase and is probably based on the sensing of Suc, and also that the structurally related disaccharide Tre can replace Suc as a regulatory compound. PMID- 10806244 TI - Purification, enzymatic characterization, and nucleotide sequence of a high isoelectric-point alpha-glucosidase from barley malt. AB - High-isoelectric-point (pI) alpha-glucosidase was purified 7, 300-fold from an extract of barley (Hordeum vulgare) malt by ammonium sulfate fractionation, ion exchange, and butyl-Sepharose chromatography. The enzyme had high activity toward maltose (k(cat) = 25 s(-1)), with an optimum at pH 4.5, and catalyzed the hydrolysis by a retaining mechanism, as shown by nuclear magnetic resonance. Acarbose was a strong inhibitor (K(i) = 1.5 microM). Molecular recognition revealed that all OH-groups in the non-reducing ring and OH-3 in the reducing ring of maltose formed important hydrogen bonds to the enzyme in the transition state complex. Mass spectrometry of tryptic fragments assigned the 92-kD protein to a barley cDNA (GenBank accession no. U22450) that appears to encode an alpha glucosidase. A corresponding sequence (HvAgl97; GenBank accession no. AF118226) was isolated from a genomic phage library using a cDNA fragment from a barley cDNA library. HvAgl97 encodes a putative 96.6-kD protein of 879 amino acids with 93.8% identity to the protein deduced from U22450. The sequence contains two active site motifs of glycoside hydrolase family 31. Three introns of 86 to 4,286 bp interrupt the coding region. The four exons vary from 218 to 1,529 bp. Gene expression analysis showed that transcription reached a maximum 48 h after the start of germination. PMID- 10806245 TI - Metabolism of methanol in plant cells. Carbon-13 nuclear magnetic resonance studies. AB - Using (13)C-NMR, we demonstrate that [(13)C]methanol readily entered sycamore (Acer pseudoplatanus L.) cells to be slowly metabolized to [3-(13)C]serine, [(13)CH(3)]methionine, and [(13)CH(3)]phosphatidylcholine. We conclude that the assimilation of [(13)C]methanol occurs through the formation of (13)CH(3)H(4)Pte glutamate (Glu)(n) and S-adenosyl-methionine, because feeding plant cells with [3 (13)CH(3)]serine, the direct precursor of (13)CH(2)H(4)Pte-Glu(n), can perfectly mimic [(13)CH(3)]methanol for folate-mediated single-carbon metabolism. On the other hand, the metabolism of [(13)C]methanol in plant cells revealed assimilation of label into a new cellular product that was identified as [(13)CH(3)]methyl-beta-D-glucopyranoside. The de novo synthesis of methyl-beta-D glucopyranoside induced by methanol did not require the formation of (13)CH(3)H(4)Pte-Glu(n) and was very likely catalyzed by a "transglycosylation" process. PMID- 10806246 TI - High-affinity potassium transport in barley roots. Ammonium-sensitive and insensitive pathways. AB - In an attempt to understand the process mediating K(+) transport into roots, we examined the contribution of the NH(4)(+)-sensitive and NH(4)(+)-insensitive components of Rb(+) transport to the uptake of Rb(+) in barley (Hordeum vulgare L.) plants grown in different ionic environments. We found that at low external Rb(+) concentrations, an NH(4)(+)-sensitive component dominates Rb(+) uptake in plants grown in the absence of NH(4)(+), while Rb(+) uptake preferentially occurs through an NH(4)(+)-insensitive pathway in plants grown at high external NH(4)(+) concentrations. A comparison of the Rb(+)-uptake properties observed in roots with those found in heterologous studies with yeast cells indicated that the recently cloned HvHAK1 K(+) transporter may provide a major route for the NH(4)(+)-sensitive component. HvHAK1 failed to complement the growth of a yeast strain defective in NH(4)(+) transport, suggesting that it could not act as an NH(4)(+) transporter. Heterologous studies also showed that the HKT1 K(+)/Na(+) cotransporter may act as a pathway for high-affinity Rb(+) transport sensitive to NH(4)(+). However, we found no evidence of an enhancement of Rb(+) uptake into roots due to Na(+) addition. The possible identity of the systems contributing to the NH(4)(+)-insensitive component in barley plants is discussed. PMID- 10806247 TI - Regulation of high-affinity nitrate transporter genes and high-affinity nitrate influx by nitrogen pools in roots of barley. AB - To investigate the regulation of HvNRT2, genes that encode high-affinity NO(3)(-) transporters in barley (Hordeum vulgare) roots, seedlings were treated with 10 mM NO(3)(-) in the presence or absence of amino acids (aspartate, asparagine, glutamate [Glu], and glutamine [Gln]), NH(4)(+), and/or inhibitors of N assimilation. Although all amino acids decreased high-affinity (13)NO(3)(-) influx and HvNRT2 transcript abundance, there was substantial interconversion of administered amino acids, making it impossible to determine which amino acid(s) were responsible for the observed effects. To clarify the role of individual amino acids, plants were separately treated with tungstate, methionine sulfoximine, or azaserine (inhibitors of nitrate reductase, Gln synthetase, and Glu synthase, respectively). Tungstate increased the HvNRT2 transcript by 20% to 30% and decreased NO(3)(-) influx by 50%, indicating that NO(3)(-) itself does not regulate transcript abundance, but may exert post-transcriptional effects. Experiments with methionine sulfoximine suggested that NH(4)(+) may down-regulate HvNRT2 gene expression and high-affinity NO(3)(-) influx by effects operating at the transcriptional and post-transcriptional levels. Azaserine decreased HvNRT2 transcript levels and NO(3)(-) influx by 97% and 95%, respectively, while decreasing Glu and increasing Gln levels. This suggests that Gln (and not Glu) is responsible for down-regulating HvNRT2 expression, although it does not preclude a contributory effect of other amino acids. PMID- 10806248 TI - Mutation of Arabidopsis plastid phosphoglucose isomerase affects leaf starch synthesis and floral initiation. AB - We isolated pgi1-1, an Arabidopsis mutant with a decreased plastid phospho glucose (Glc) isomerase activity. While pgi1-1 mutant has a deficiency in leaf starch synthesis, it accumulates starch in root cap cells. It has been shown that a plastid transporter for hexose phosphate transports cytosolic Glc-6-P into plastids and expresses restricted mainly to the heterotrophic tissues. The decreased starch content in leaves of the pgi1-1 mutant indicates that cytosolic Glc-6-P cannot be efficiently transported into chloroplasts to complement the mutant's deficiency in chloroplastic phospho-Glc isomerase activity for starch synthesis. We cloned the Arabidopsis PGI1 gene and showed that it encodes the plastid phospho-Glc isomerase. The pgi1-1 allele was found to have a single nucleotide substitution, causing a Ser to Phe transition. While the flowering times of the Arabidopsis starch-deficient mutants pgi1, pgm1, and adg1 were similar to that of the wild type under long-day conditions, it was significantly delayed under short-day conditions. The pleiotropic phenotype of late flowering conferred by these starch metabolic mutations suggests that carbohydrate metabolism plays an important role in floral initiation. PMID- 10806249 TI - Purification and characterization of a novel pumpkin short-chain acyl-coenzyme A oxidase with structural similarity to acyl-coenzyme A dehydrogenases. AB - A novel pumpkin (Cucurbita pepo) short-chain acyl-coenzyme A (CoA) oxidase (ACOX) was purified to homogeneity by hydrophobic-interaction, hydroxyapatite, affinity, and anion-exchange chromatography. The purified enzyme is a tetrameric protein, consisting of apparently identical 47-kD subunits. The protein structure of this oxidase differs from other plant and mammalian ACOXs, but is similar to the protein structure of mammalian mitochondrial acyl-CoA dehydrogenase (ACDH) and the recently identified plant mitochondrial ACDH. Subcellular organelle separation by sucrose density gradient centrifugation revealed that the enzyme is localized in glyoxysomes, whereas no immunoreactive bands of similar molecular weight were detected in mitochondrial fractions. The enzyme selectively catalyzes the oxidation of CoA esters of fatty acids with 4 to 10 carbon atoms, and exhibits the highest activity on C-6 fatty acids. Apparently, the enzyme has no activity on CoA esters of branched-chain or dicarboxylic fatty acids. The enzyme is slightly inhibited by high concentrations of substrate and it is not inhibited by Triton X-100 at concentrations up to 0.5% (v/v). The characteristics of this novel ACOX enzyme are discussed in relation to other ACOXs and ACDHs. PMID- 10806250 TI - Ascorbate biosynthesis in mitochondria is linked to the electron transport chain between complexes III and IV. AB - Ascorbic acid is synthesized from galactono-gamma-lactone (GL) in plant tissues. An improved extraction procedure involving ammonium sulfate precipitation of membrane proteins from crude leaf homogenates yielded a simple, quick method for determining tissue activities of galactono-gamma-lactone dehydrogenase (GLDH). Total foliar ascorbate and GLDH activity decreased with leaf age. Subcellular fractionation experiments using marker enzymes demonstrated that 80% of the total GLDH activity was located on the inner mitochondrial membrane, and 20% in the microsomal fraction. Specific antibody raised against potato (Solanum tuberosum L.) tuber GLDH recognized a 56-kD polypeptide in extracts from the mitochondrial membranes but failed to detect the equivalent polypeptide in microsomes. We demonstrate that isolated intact mitochondria synthesize ascorbate in the presence of GL. GL stimulated mitochondrial electron transport rates. The respiration inhibitor antimycin A stimulated ascorbate biosynthesis, while cyanide inhibited both respiration and ascorbate production. GL-dependent oxygen uptake was observed in isolated intact mitochondria. This evidence suggests that GLDH delivers electrons to the mitochondrial electron transport chain between complexes III and IV. PMID- 10806251 TI - Regulation of carbonic anhydrase expression by zinc, cobalt, and carbon dioxide in the marine diatom Thalassiosira weissflogii. AB - TWCA1 is the major Zn-requiring isoform of carbonic anhydrase (CA) in the marine diatom Thalassiosira weissflogii. We have examined the roles that trace metals and CO(2) play in the regulation of TWCA1 expression over ranges of concentrations that bracket those encountered in the marine environment. Both steady-state levels of TWCA1 and the kinetics of induction were measured by western analysis. TWCA1 levels correlated well with cellular CA activity levels. TWCA1 was induced at a low CO(2) concentration but the level of induction, as determined by western analysis, was dependent on the availability of Zn. Co effectively substituted for Zn in regulating TWCA1 expression and promoting TWCA1 activity. Upon shift from low to high CO(2), the concentration of TWCA1 decreased. The expression of TWCA1 is diel cycle regulated, and cellular TWCA1 decreased during the dark phase. These results provide the basis for studying the expression of CA in field populations and, taken together with previous radiolabeling studies, provide strong evidence of in vivo metal substitution of Co for Zn in a CA. Our data also support the conclusion that TWCA1 plays a central role in carbon acquisition in T. weissflogii. PMID- 10806252 TI - AVP2, a sequence-divergent, K(+)-insensitive H(+)-translocating inorganic pyrophosphatase from Arabidopsis. AB - Plant vacuolar H(+)-translocating inorganic pyrophosphatases (V-PPases; EC 3.6.1.1) have been considered to constitute a family of functionally and structurally monotonous intrinsic membrane proteins. Typified by AVP1 (V. Sarafian, Y. Kim, R.J. Poole, P.A. Rea [1992] Proc Natl Acad Sci USA 89: 1775 1779) from Arabidopsis, all characterized plant V-PPases share greater than 84% sequence identity and catalyze K(+)-stimulated H(+) translocation. Here we describe the molecular and biochemical characterization of AVP2 (accession no. AF182813), a sequence-divergent (36% identical) K(+)-insensitive, Ca(2+) hypersensitive V-PPase active in both inorganic pyrophosphate hydrolysis and H(+) translocation. The differences between AVP2 and AVP1 provide the first indication that plant V-PPases from the same organism fall into two distinct categories. Phylogenetic analyses of these and other V-PPase sequences extend this principle by showing that AVP2, rather than being an isoform of AVP1, is but one representative of a novel category of AVP2-like (type II) V-PPases that coexist with AVP1-like (type I) V-PPases not only in plants, but also in apicomplexan protists such as the malarial parasite Plasmodium falciparum. PMID- 10806253 TI - Fruit-localized phytochromes regulate lycopene accumulation independently of ethylene production in tomato. AB - We show that phytochromes modulate differentially various facets of light-induced ripening of tomato fruit (Solanum lycopersicum L.). Northern analysis demonstrated that phytochrome A mRNA in fruit accumulates 11.4-fold during ripening. Spectroradiometric measurement of pericarp tissues revealed that the red to far-red ratio increases 4-fold in pericarp tissues during ripening from the immature-green to the red-ripe stage. Brief red-light treatment of harvested mature-green fruit stimulated lycopene accumulation 2. 3-fold during fruit development. This red-light-induced lycopene accumulation was reversed by subsequent treatment with far-red light, establishing that light-induced accumulation of lycopene in tomato is regulated by fruit-localized phytochromes. Red-light and red-light/far-red-light treatments during ripening did not influence ethylene production, indicating that the biosynthesis of this ripening hormone in these tissues is not regulated by fruit-localized phytochromes. Compression analysis of fruit treated with red light or red/far-red light indicated that phytochromes do not regulate the rate or extent of pericarp softening during ripening. Moreover, treatments with red or red/far-red light did not alter the concentrations of citrate, malate, fructose, glucose, or sucrose in fruit. These results are consistent with two conclusions: (a) fruit-localized phytochromes regulate light-induced lycopene accumulation independently of ethylene biosynthesis; and (b) fruit-localized phytochromes are not global regulators of ripening, but instead regulate one or more specific components of this developmental process. PMID- 10806254 TI - Radiotracer and computer modeling evidence that phospho-base methylation is the main route of choline synthesis in tobacco. AB - Among flowering plants, the synthesis of choline (Cho) from ethanolamine (EA) can potentially occur via three parallel, interconnected pathways involving methylation of free bases, phospho-bases, or phosphatidyl-bases. We investigated which pathways operate in tobacco (Nicotiana tabacum L.) because previous work has shown that the endogenous Cho supply limits accumulation of glycine betaine in transgenic tobacco plants engineered to convert Cho to glycine betaine. The kinetics of metabolite labeling were monitored in leaf discs supplied with [(33)P]phospho-EA, [(33)P]phospho-monomethylethanolamine, or [(14)C]formate, and the data were subjected to computer modeling. Because partial hydrolysis of phospho-bases occurred in the apoplast, modeling of phospho-base metabolism required consideration of the re-entry of [(33)P]phosphate into the network. Modeling of [(14)C]formate metabolism required consideration of the labeling of the EA and methyl moieties of Cho. Results supported the following conclusions: (a) The first methylation step occurs solely at the phospho-base level; (b) the second and third methylations occur mainly (83%-92% and 65%-85%, respectively) at the phospho-base level, with the remainder occurring at the phosphatidyl-base level; and (c) free Cho originates predominantly from phosphatidylcholine rather than from phospho-Cho. This study illustrates how computer modeling of radiotracer data, in conjunction with information on chemical pool sizes, can provide a coherent, quantitative picture of fluxes within a complex metabolic network. PMID- 10806255 TI - Integrated temporal regulation of the photorespiratory pathway. Circadian regulation of two Arabidopsis genes encoding serine hydroxymethyltransferase. AB - The photorespiratory pathway is comprised of enzymes localized within three distinct cellular compartments: chloroplasts, peroxisomes, and mitochondria. Photorespiratory enzymes are encoded by nuclear genes, translated in the cytosol, and targeted into these distinct subcellular compartments. One likely means by which to regulate the expression of the genes encoding photorespiratory enzymes is coordinated temporal control. We have previously shown in Arabidopsis that a circadian clock regulates the expression of the nuclear genes encoding both chloroplastic (Rubisco small subunit and Rubisco activase) and peroxisomal (catalase) components of the photorespiratory pathway. To determine whether a circadian clock also regulates the expression of genes encoding mitochondrial components of the photorespiratory pathway, we characterized a family of Arabidopsis serine hydroxymethyltransferase (SHM) genes. We examined mRNA accumulation for two of these family members, including one probable photorespiratory gene (SHM1) and a second gene expressed maximally in roots (SHM4), and show that both exhibit circadian oscillations in mRNA abundance that are in phase with those described for other photorespiratory genes. In addition, we show that SHM1 mRNA accumulates in light-grown seedlings, although this response is probably an indirect consequence of the induction of photosynthesis and photorespiration by illumination. PMID- 10806256 TI - The yeast HAL1 gene improves salt tolerance of transgenic tomato. AB - Overexpression of the HAL1 gene in yeast has a positive effect on salt tolerance by maintaining a high internal K(+) concentration and decreasing intracellular Na(+) during salt stress. In the present work, the yeast gene HAL1 was introduced into tomato (Lycopersicon esculentum Mill.) by Agrobacterium tumefaciens-mediated transformation. A sample of primary transformants was self-pollinated, and progeny from both transformed and non-transformed plants (controls) were evaluated for salt tolerance in vitro and in vivo. Results from different tests indicated a higher level of salt tolerance in the progeny of two different transgenic plants bearing four copies or one copy of the HAL1 gene. In addition, measurement of the intracellular K(+) to Na(+) ratios showed that transgenic lines were able to retain more K(+) than the control under salt stress. Although plants and yeast cannot be compared in an absolute sense, these results indicate that the mechanism controlling the positive effect of the HAL1 gene on salt tolerance may be similar in transgenic plants and yeast. PMID- 10806257 TI - Regulation of a plant SNF1-related protein kinase by glucose-6-phosphate. AB - One of the major protein kinases (PK(III)) that phosphorylates serine-158 of spinach sucrose-phosphate synthase (SPS), which is responsible for light/dark modulation of activity, is known to be a member of the SNF1-related family of protein kinases. In the present study, we have developed a fluorescence-based continuous assay for measurement of PK(III) activity. Using the continuous assay, along with the fixed-time-point (32)P-incorporation assay, we demonstrate that PK(III) activity is inhibited by glucose-6-phosphate (Glc-6-P). Relative inhibition by Glc-6-P was increased by decreasing pH from 8. 5 to 5.5 and by reducing the concentration of Mg(2+) in the assay from 10 to 2 mM. Under likely physiological conditions (pH 7.0 and 2 mM Mg(2+)), 10 mM Glc-6-P inhibited kinase activity approximately 70%. Inhibition by Glc-6-P could not be ascribed to contaminants in the commercial preparations. Other metabolites inhibited PK(III) in the following order: Glc-6-P > mannose-6-P, fructose-1,6P(2) > ribose-5-P, 3 PGA, fructose-6-P. Inorganic phosphate, Glc, and AMP were not inhibitory, and free Glc did not reverse the inhibition by Glc-6-P. Because SNF1-related protein kinases are thought to function broadly in the regulation of enzyme activity and gene expression, Glc-6-P inhibition of PK(III) activity potentially provides a mechanism for metabolic regulation of the reactions catalyzed by these important protein kinases. PMID- 10806258 TI - Thermal balloon and rollerball ablation to treat menorrhagia: two-year results of a multicenter, prospective, randomized, clinical trial. AB - STUDY OBJECTIVE: To evaluate 2-year follow-up results in patients participating in a randomized, clinical trial comparing uterine balloon therapy with rollerball endometrial ablation. STUDY DESIGN: Prospective, randomized, clinical trial (Canadian Task Force classification I). SETTING: Fourteen university-affiliated and private practice sites. PATIENTS: Two hundred fifty-five women with menorrhagia. INTERVENTIONS: Patients were randomized to rollerball or uterine balloon endometrial ablation. MEASUREMENTS AND MAIN RESULTS: Patient satisfaction with both treatments was consistently high. Only 15 hysterectomies had been performed (6 for menorrhagia) at the end of 2 years, 11 in the rollerball group, 4 in the balloon therapy group. CONCLUSION: Endometrial ablation by both procedures was highly successful in avoiding hysterectomy and relieving symptoms of menorrhagia. Additional benefits were reduction in dysmenorrhea and premenstrual syndrome. PMID- 10806259 TI - Pain referral patterns in the pelvis. AB - STUDY OBJECTIVE: To determine if right-left pelvic pain orientation exists in all patients, and to estimate the frequency of pain referral patterns in the pelvis. DESIGN: Prospective, nonrandomized trial (Canadian Task Force classification II 1). SETTING: University-affiliated hospital. PATIENTS: One hundred consecutive women with no pain undergoing tubal sterilization or investigation of infertility (group A) were compared with 225 women with chronic pelvic pain of greater than 6 months' duration (group B). INTERVENTION: Patient-assisted laparoscopy with intravenous conscious sedation and pelvic pain mapping. MEASUREMENTS AND MAIN RESULTS: Of women in group A, 69% had correct right-left orientation, 18% had pain perceived on the opposite side of the abdomen, and 31% had pain referred to another location in the abdomen. The results were similar in women in group B, 65% of whom had correct orientation, 15% had pain perceived on the opposite side of the abdomen, and 35% had pain referred to another location. CONCLUSION: This demonstrates the need to ask on which side a woman feels pain during physical examination. It is not correct to assume that an answer of "yes" means the same side of the pelvis that is being examined. Patient-assisted laparoscopy and pelvic pain mapping are excellent in correlating the symptom with the pathology. PMID- 10806260 TI - Hysteroscopic evaluation of the endometrium in postmenopausal women taking tamoxifen. AB - STUDY OBJECTIVE: To evaluate hysteroscopic endometrial changes due to tamoxifen therapy in postmenopausal women with breast cancer. DESIGN: Retrospective study (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. Patients. Eighty-eight postmenopausal women (or with iatrogenic amenorrhea) receiving tamoxifen 20 mg/day for at least 1 year for breast cancer. INTERVENTION: Record review of patients undergoing transvaginal sonography (TVS) and office hysteroscopy with eye-directed biopsy specimens obtained with a 5-mm, continuous-flow, operative hysteroscope. MEASUREMENTS AND MAIN RESULTS: Patients with thickened endometrium and pathologic findings at hysteroscopy had taken tamoxifen for significantly longer times than those without such findings (p < 0.05). CONCLUSION: Our findings confirm the estrogenic effect of tamoxifen on endometrium. Endometrial evaluation by TVS suggests further diagnostic procedures, but only hysteroscopy allows the surgeon to visualize endometrial lesions and obtain eye-directed biopsy tissue. PMID- 10806261 TI - Hysteroscopic evaluation of menopausal women with endometrial thickness of 4 mm or more. AB - STUDY OBJECTIVE: To assess the diagnostic and operative potential of hysteroscopy in postmenopausal patients selected by ultrasound criteria. DESIGN: Cohort study (Canadian Task Force classification II-2). SETTING: Outpatient ultrasound and hysteroscopy department of a university-affiliated hospital. PATIENTS: One hundred fifty-five postmenopausal women with endometrial thickness of 4 mm or more by ultrasound, in menopause for at least 1 year, with or without menopausal complaints. INTERVENTIONS: Transvaginal ultrasound and office hysteroscopy with eye-directed biopsy specimens using a 5-mm, continuous-flow, operative hysteroscope. MEASUREMENTS AND MAIN RESULTS: Of the 155 women, 129 (83%) were asymptomatic (irregular bleeding). Hysteroscopy showed endometrial pathology in 28% of asymptomatic patients (23 polyps, 5 cases of hyperplasia, 8 submucous myomata) and 76% of symptomatic women (13 polyps, 6 hyperplasia, 1 submucous myoma). Hysteroscopic results compared with histologic diagnosis showed a positive predictive value equal to 97. 1% and 95% in asymptomatic and symptomatic women, respectively, and a negative predictive value equal to 100% in both groups. CONCLUSION: Office hysteroscopy with endometrial biopsy samples has a diagnostic and operative role in postmenopausal patients selected based on endometrial thickness on ultrasound, in view of the high prevalence of endometrial pathology in both symptomatic and asymptomatic women. PMID- 10806262 TI - Hysteroscopic evaluation of menopausal patients with sonographically atrophic endometrium. AB - STUDY OBJECTIVE: To assess the diagnostic and operative potential of hysteroscopy in postmenopausal patients selected by ultrasound criteria. DESIGN: Prospective evaluation (Canadian Task Force classification II-2). SETTING: Outpatient ultrasound and hysteroscopy department of a university-affiliated hospital. PATIENTS: Two hundred twelve women with an endometrial thickness less than 4 mm on ultrasound and in menopause for at least 1 year. INTERVENTIONS: Transvaginal ultrasound and office hysteroscopy, with eye-directed biopsy specimens obtained with a 5-mm, continuous-flow operative hysteroscope, and performed without anesthesia. MEASUREMENTS AND MAIN RESULTS: Only 13 (6%) patients were symptomatic (irregular bleeding). Hysteroscopic diagnosis of endometrial polyps in three women (23%) was confirmed by histology. In the remaining 199 (94%) asymptomatic patients with atrophic endometrium on ultrasound, hysteroscopy showed an endometrial pathology in 10% (16 polyps, 4 submucous myomas); in one patient histologic evaluation disclosed focal adenocarcinoma in an endometrial polyp. CONCLUSION: Hysteroscopy allows a proper histologic diagnosis, even in asymptomatic postmenopausal women with atrophic endometrium on ultrasound. PMID- 10806263 TI - Accuracy of preoperative diagnostic tools and outcome of hysteroscopic management of menstrual dysfunction. AB - STUDY OBJECTIVES: To review diagnoses, complications, and surgical findings in women treated for abnormal uterine bleeding by operative hysteroscopy, and to assess the accuracy of preoperative transvaginal ultrasound (TVS), saline infusion sonography (SIS), diagnostic hysteroscopy, and endometrial biopsy. DESIGN: Retrospective study (Canadian task force classification II-2). SETTING: Tertiary care academic medical center. PATIENTS: Three hundred seventy-five women. Intervention. Operative hysteroscopy for abnormal uterine bleeding. MEASUREMENTS AND MAIN RESULTS: Main indications for hysteroscopy were postmenopausal bleeding (164 patients, 43.7%) and abnormal premenopausal uterine bleeding (211, 56.3%). Main pathology findings were endometrial polyps (172, 45.9%) and submucous myomas (105, 28%). Polyps had histologic abnormalities in 18 patients. Sensitivity of preoperative diagnostic tools for all intrauterine abnormalities and specifically for myomas and polyps was TVS 74% and 39%, SIS 96% and 96%, hysteroscopy 100% and 99%, and Pipelle biopsy 24% and 10%. The complication rate was 1.3%. Postmenopausal women felt significantly more improvement in symptoms (p = 0.02), and were more satisfied (p or =2) and non-phototoxic (PI(clin)<2) FQ could be completely discriminated using these parameters, and that the in vitro models were able to rank the relative phototoxic potential of the eight FQ. PMID- 10806380 TI - High yield purification and physico-chemical properties of full-length recombinant allelic variants of sheep prion protein linked to scrapie susceptibility. AB - Sheep susceptibility to scrapie is governed by polymorphisms at two major sites, codons 136 and 171, of the prp gene. To get more insight into the prion protein (PrP) sequence-linked basis of differential scrapie susceptibility, a high yield one-step method for the purification (over 99% final purity) of the full-length recombinant sheep PrP was developed, based on the affinity of the conserved octapeptide repeats for transition-metal cations. Thermal and chemical denaturation experiments and limited proteolysis studies were performed on the natural variants (A136R171, V136Q171 and A136Q171) and a recombinant PrP mutated at position 136 (V136R171). Results revealed the influence of mutations in positions 136 and 171 on the folding thermodynamic parameters and on the conformation of the C-terminal domain. Together, our results show that the VQ cellular protein linked to higher scrapie susceptibility is intrinsically more compact and/or stable than the resistance-linked AR counterpart. This might lead to a lower in vivo clearance rate of VQ and a consequently higher probability of occurrence of pathological events. PMID- 10806381 TI - The heme-independent manganese-peroxidase activity depends on the presence of the C-terminal domain within the Streptomyces reticuli catalase-peroxidase CpeB. AB - Streptomyces reticuli produces a heme-containing homodimeric enzyme (160 kDa), the catalase-peroxidase CpeB, which is processed to the enzyme CpeC during prolonged growth. CpeC contains four subunits of 60 kDa each that do not include the C-terminal portion of the progenitor subunits. A genetically engineered cpeB gene encodes a truncated subunit lacking 195 of the C-terminal amino acids; four of these subunits assemble to form the enzyme CpeD. Heme binds most strongly in CpeB, least in CpeD. The catalase-peroxidase CpeB and its apo-form (obtained after extraction of heme) catalyze the peroxidation of Mn(II) to Mn(III), independent of the presence or absence of the heme inhibitor KCN. CpeC and CpeD, in contrast, do not exhibit manganese-peroxidase activity. The data show for the first time that a bacterial catalase-peroxidase has a heme-independent manganese peroxidase activity, which depends on the presence of the C-terminal domain. PMID- 10806382 TI - Polytomella spp. growth on ethanol. Extracellular pH affects the accumulation of mitochondrial cytochrome c550. AB - A defined medium with ethanol as sole carbon source was devised for growth of the colorless, unicellular alga Polytomella spp. Cell density on this carbon source was related to extracellular pH. An acidic pH was required for ethanol utilization; best yields were obtained at pH 3.7. Spectroscopic analysis of the cells showed that the concentration of cytochrome c per cell was 40% higher than at pH 6.0; the concentrations of cytochrome a606 (cytochrome c oxidase) and b566 (cytochrome bc1 complex) were the same. A soluble cytochrome c550 was purified from cells grown at pH 3.7 and characterized by peptide sequencing as the 12-kDa cytochrome c550 of the mitochondrial respiratory chain. Immunoblots of total cell proteins showed higher accumulation of cytochrome c550 at pH 3.7 than at pH 6.0. RNA blot analysis gave clear evidence of the abundance of c550 transcript in cells grown at pH 3.7. The amount of mitochondrial proteins obtained from cells grown at pH 3.7 was twofold higher than that of cells grown at pH 6.0. Mitochondria isolated from both cell types readily oxidized succinate, malate or ethanol. The rates of oxygen uptake were 20-25% higher in mitochondria from cells grown at pH 3.7. Cyanide and antimycin A inhibited respiration with succinate up to 95% in both types of mitochondria. The participation of cytochrome c550 in mitochondrial electron transport from succinate to oxygen was shown by spectral measurements. PMID- 10806383 TI - Stability and folding of the protein complexes of barnase. AB - Native-like complexes of proteins, formed by the association of two complementary fragments comprising the entire sequence of the protein, can be used to gain insight into the stability and folding of the intact protein. We have studied the structural, thermodynamic and kinetic properties of four barnase complexes, with the cleavage site at different positions of the amino-acid chain (CB36, at position 36; CB56, at position 56; CB68, at position 68; and CB79, at position 79). The four barnase complexes have native-like structure as shown by fluorescence, far-and near-UV CD, size-exclusion chromatography and NMR. The NMR characterization indicated that the structural changes were mainly located in regions close to the cleavage site. The main core of the protein was fully formed and the overall structure was similar to that of intact barnase. The thermal and chemical denaturation showed that all complexes were substantially destabilized. CB56 displayed two denaturation transitions, probably because of the presence of partially folded conformations around the cleavage site. The rate constant for the association/folding of fragments decreased with the decreasing length of the C-terminal fragment. Thus, the larger the fragment (and, consequently, the larger the amount of residual native-like structure), the faster the association. These findings are consistent with the proposed model of barnase folding. PMID- 10806384 TI - Proteomic analysis of the Escherichia coli outer membrane. AB - Outer membrane proteins (OMPs) of Gram-negative bacteria are key molecules that interface the cell with the environment. Traditional biochemical and genetic approaches have yielded a wealth of knowledge relating to the function of OMPs. Nonetheless, with the completion of the Escherichia coli genome sequencing project there is the opportunity to further expand our understanding of the organization, expression and function of the OMPs in this Gram-negative bacterium. In this report we describe a proteomic approach which provides a platform for parallel analysis of OMPs. We propose a rapid method for isolation of bacterial OMPs using carbonate incubation, purification and protein array by two-dimensional electrophoresis, followed by protein identification using mass spectrometry. Applying this method to examine E. coli K-12 cells grown in minimal media we identified 21 out of 26 (80%) of the predicted integral OMPs that are annotated in SWISS-PROT release 37 and predicted to separate within the range of pH 4-7 and molecular mass 10-80 kDa. Five outer membrane lipoproteins were also identified and only minor contamination by nonmembrane proteins was observed. Importantly, this research readily demonstrates that integral OMPs, commonly missing from 2D gel maps, are amenable to separation by two-dimensional electrophoresis. Two of the identified OMPs (YbiL, YeaF) were previously known only from their ORFs, and their identification confirms the cognate genes are transcribed and translated. Furthermore, we show that like the E. coli iron receptors FhuE and FhuA, the expression of YbiL is markedly increased by iron limitation, suggesting a putative role for this protein in iron transport. In an additional demonstration we show the value of parallel protein analysis to document changes in E. coli OMP expression as influenced by culture temperature. PMID- 10806385 TI - Interaction between the lipoamide-containing H-protein and the lipoamide dehydrogenase (L-protein) of the glycine decarboxylase multienzyme system. 1. Biochemical studies. AB - Lipoamide dehydrogenase or dihydrolipoamide dehydrogenase (EC 1.8.1. 4) is the E3 protein component of the mitochondrial 2-oxoacid dehydrogenase multienzyme complexes. It is also the L-protein component of the glycine decarboxylase system. Although the enzymology of this enzyme has been studied exhaustively using free lipoamide as substrate, no data are available concerning the kinetic parameters of this enzyme with its physiological substrates, the dihydrolipoyl domain of the E2 component (dihydrolipoyl acyltransferase) of the 2-oxoacid dehydrogenase multienzyme complexes or the dihydrolipoyl H-protein of the mitochondrial glycine decarboxylase. In this paper, we demonstrate that Tris(2 carboxyethyl)phosphine, a specific disulfide reducing agent, allows a continuous reduction of the lipoyl group associated with the H-protein during the course of the reaction catalysed by the L-protein. This provided a valuable new tool with which to study the catalytic properties of the lipoamide dehydrogenase. The L protein displayed a much higher affinity for the dihydrolipoyl H-protein than for free dihydrolipoamide. The oxidation of the dihydrolipoyl H-protein was not affected by the presence of structurally related analogues (apoH-protein or octanoylated H-protein). In marked contrast, these analogues strongly and competitively inhibited the decarboxylation of the glycine molecule catalysed by the P-protein component of the glycine decarboxylase system. Small unfolded proteolytic fragments of the H-protein, containing the lipoamide moiety, displayed Km values for the L-protein close to that found for the H-protein. On the other hand, these fragments were not able to promote the decarboxylation of the glycine in the presence of the P-protein. New highly hydrophilic lipoate analogues were synthesized. All of them showed Km and kcat/Km values very close to that found for the H-protein. From our results we concluded that no structural interaction is required for the L-protein to catalyse the oxidation of the dihydrolipoyl H-protein. We discuss the possibility that one function of the H protein is to maintain a high concentration of the hydrophobic lipoate molecules in a nonmicellar state which would be accessible to the catalytic site of the lipoamide dehydrogenase. PMID- 10806386 TI - Interaction between the lipoamide-containing H-protein and the lipoamide dehydrogenase (L-protein) of the glycine decarboxylase multienzyme system 2. Crystal structures of H- and L-proteins. AB - The glycine decarboxylase complex consists of four different component enzymes (P , H-, T- and L-proteins). The 14-kDa lipoamide-containing H-protein plays a pivotal role in the complete sequence of reactions as its prosthetic group (lipoic acid) interacts successively with the three other components of the complex and undergoes a cycle of reductive methylamination, methylamine transfer and electron transfer. With the aim to understand the interaction between the H protein and its different partners, we have previously determined the crystal structure of the oxidized and methylaminated forms of the H-protein. In the present study, we have crystallized the H-protein in its reduced state and the L protein (lipoamide dehydrogenase or dihydrolipoamide dehydrogenase). The L protein has been overexpressed in Escherichia coli and refolded from inclusion bodies in an active form. Crystals were obtained from the refolded L-protein and the structure has been determined by X-ray crystallography. This first crystal structure of a plant dihydrolipoamide dehydrogenase is similar to other known dihydrolipoamide dehydrogenase structures. The crystal structure of the H-protein in its reduced form has been determined and compared to the structure of the other forms of the protein. It is isomorphous to the structure of the oxidized form. In contrast with methylaminated H-protein where the loaded lipoamide arm was locked into a cavity of the protein, the reduced lipoamide arm appeared freely exposed to the solvent. Such a freedom is required to allow its targeting inside the hollow active site of L-protein. Our results strongly suggest that a direct interaction between the H- and L-proteins is not necessary for the reoxidation of the reduced lipoamide arm bound to the H-protein. This hypothesis is supported by biochemical data [Neuburger, M., Polidori, A.M., Pietre, E., Faure, M., Jourdain, A., Bourguignon, J., Pucci, B. & Douce, R. (2000) Eur. J. Biochem. 267, 2882-2889] and by small angle X-ray scattering experiments reported herein. PMID- 10806387 TI - Three pheromone-binding proteins in olfactory sensilla of the two silkmoth species Antheraea polyphemus and Antheraea pernyi. AB - Females of the sibling silkmoth species Antheraea polyphemus and A. pernyi use the same three sex pheromone components in different ratios to attract conspecific males. Accordingly, the sensory hairs on the antennae of males contain three receptor cells sensitive to each of the pheromone components. In agreement with the number of pheromones used, three different pheromone-binding proteins (PBPs) could be identified in pheromone-sensitive hairs of both species by combining biochemical and molecular cloning techniques. MALDI-TOF MS of sensillum lymph droplets from pheromone-sensitive sensilla trichodea of male A. polyphemus revealed the presence of three major peaks with m/z of 15702, 15752 and 15780 and two minor peaks of m/z 15963 and 15983. In Western blots with four antisera raised against different silkmoth odorant-binding proteins, immunoreactivity was found only with an anti-(Apol PBP) serum. Free-flow IEF, ion exchange chromatography and Western blot analyses revealed at least three anti (Apol PBP) immunoreactive proteins with pI values between 4.4 and 4.7. N-Terminal sequencing of these three proteins revealed two proteins (Apol PBP1a and Apol PBP1b) identical in the first 49 amino acids to the already known PBP (Apol PBP1) [Raming, K. , Krieger, J. & Breer, H. (1989) FEBS Lett. 256, 2215-2218] and a new PBP having only 57% identity with this amino-acid region. Screening of antennal cDNA libraries with an oligonucleotide probe corresponding to the N-terminal end of the new A. polyphemus PBP, led to the discovery of full length clones encoding this protein in A. polyphemus (Apol PBP3) and in A. pernyi (Aper PBP3). By screening the antennal cDNA library of A. polyphemus with a digoxigenin-labelled A. pernyi PBP2 cDNA [Krieger, J., Raming, K. & Breer, H. (1991) Biochim. Biophys. Acta 1088, 277-284] a homologous PBP (Apol PBP2) was cloned. Binding studies with the two main pheromone components of A. polyphemus and A. pernyi, the (E,Z)-6, 11 hexadecadienyl acetate (AC1) and the (E,Z)-6,11-hexadecadienal (ALD), revealed that in A. polyphemus both Apol PBP1a and the new Apol PBP3 bound the 3H-labelled acetate, whereas no binding of the 3H-labelled aldehyde was found. In A. pernyi two PBPs from sensory hair homogenates showed binding affinity for the AC1 (Aper PBP1) and the ALD (Aper PBP2), respectively. PMID- 10806388 TI - Curvature properties of novel forms of phosphatidylcholine with branched acyl chains. AB - We studied the properties of a series of phosphatidylcholine molecules with branched acyl chains. These lipids have previously been shown to have marked stimulatory effects on the side-chain cleavage activity of cytochrome P450SCC (CYP11A1), an enzyme of the inner mitochondrial membrane. The synthetic lipids used were diacyl phosphatidylcholines with the decanoyl, dodecanoyl or tetradecanoyl chain having a hexyl, octyl or decyl straight chain aliphatic branch at the 2-position. All three lipids lowered the bilayer to hexagonal phase transition temperature of dielaidoyl phosphatidylethanolamine, the lipids with longer acyl chains being more effective in this regard. As pure lipids all of the forms were found by X-ray diffraction to be predominantly in the hexagonal phase (HII) over the entire temperature range of 7-75 degrees C. The properties of the HII phase were unusual with regard to the small size of the lattice spacings and the small temperature dependence of the spacings. We used tetradecane to relieve hydrocarbon packing constraints to determine the intrinsic radius of curvature of the lipid monolayer. The elastic bending modulus was measured in the presence of tetradecane by introducing an osmotic gradient across the hexagonal phase cylinders with aqueous solutions of poly(ethylene glycol). The elastic bending modulus was found to be higher than that observed with other lipids and to increase with temperature. Both the small intrinsic radius of curvature and the high elastic bending modulus indicate that the presence of these lipids in bilayer membranes will impose a high degree of negative curvature strain. PMID- 10806389 TI - Flexibility and stability of the structure of cytochromes P450 3A4 and BM-3. AB - The flexibility of the structure and compressibility of the respective active site of cytochromes P450 3A4 (CYP3A4) and BM-3 (CYP102) were studied using absorption spectroscopy in the ultraviolet and visual regions. Conformational changes in the overall protein structures of both CYP3A4 and CYP102 due to the effects of temperature and pressure are reversible. However, the enzymes differ in the properties of their active sites. The CYP3A4 enzyme denatures to the inactive P420 form relatively easy, at 3000 bar over half is converted to P420. The compressibility of its active site is lower than that of CYP102 and is greater with the substrate bound, which is in line with the observed lack of a stabilizing effect of the substrate on its conformation under pressure. In contrast, CYP102, although having the most compressible active site among the P450s, possesses a structure that does not denature easily to the inactive (P420) form under pressure. In this respect, it resembles the P450 isolated from acidothermophilic archaebacteria [McLean, M.A., Maves, S.A., Weiss, K.E., Krepich, S. & Sligar, S.G. (1998) Biochem. Biophys. Res. Commun. 252, 166-172]. PMID- 10806390 TI - ARP3beta, the gene encoding a new human actin-related protein, is alternatively spliced and predominantly expressed in brain neuronal cells. AB - A cDNA encoding a new human actin-related protein (ARP) was cloned. The corresponding protein is highly conserved with the previously described ARP3 protein, suggesting that it represents a second isoform of the human ARP3 subfamily. This new actin-related protein was subsequently named ARP3beta and represents the second example of multiple isoforms of an actin-related protein in a single organism. The ARP3beta gene was mapped to chromosome band 7q34, centromeric to Sonic Hedgehog. Gene structure analysis revealed that at least part of the observed ARP3beta mRNA heterogeneity is caused by alternative splicing resulting in exon skipping. Transcripts produced after exon 2 skipping are predicted to encode truncated products, whose functionality is still unclear. An ARP3beta pseudogene was detected on chromosome 2p11 by database searching. Several ARP3beta mRNA species were detected by Northern blotting and their abundance varied importantly among tissues: the highest expression levels were detected in fetal and adult brain, whereas lower levels were observed in liver, muscle and pancreas. In contrast, ARP3 mRNAs were detected in all tissues tested. Using in situ hybridization, the expression of ARP3beta in brain was shown to be restricted to neurons and epithelial cells from choroid plexus. This suggests a specific function for ARP3beta in the physiology of the development and/or maintenance of distinct subsets of nerve cells. PMID- 10806391 TI - Solution structure of ileal lipid binding protein in complex with glycocholate. AB - Ileal lipid binding protein (ILBP) is a cytosolic lipid-binding protein that binds both bile acids and fatty acids. We have determined the solution structure of porcine ILBP in complex with glycocholate by homonuclear and heteronuclear two dimensional NMR spectroscopy. The conformation of the protein-ligand complex was determined by restrained energy minimization and simulated annealing calculations after docking the glycocholate ligand into the protein structure. The overall tertiary structure of ILBP is highly analogous to the three-dimensional structures of several other intracellular lipid binding proteins (LBPs). Like the apo-structure, the bile-acid complex of ILBP is composed of 10 anti-parallel beta strands that form a water-filled clam-shell structure, and two short alpha helices. Chemical shift data indicated that the bile acid ligand is bound inside the protein cavity. Furthermore, 13C-edited heteronuclear single-quantum correlation-NOESY experiments showed NOE contacts between several aromatic residues located in the proposed bile acid portal region and the 13C-labeled ligand. A single bile acid molecule is bound inside the protein, with the steroid moiety penetrating deep into the water-accessible internal cavity, such that ring A is located right above the plane of the Trp49 indole ring. The carboxylate tail of the ligand is protruding from the proposed bile acid portal into the surrounding aqueous solution. The body of the steroid moiety is oriented with the nonpolar face in contact with the mostly hydrophobic residues of beta-strands C, D and E, while the polar face shows contacts with the side-chains of Tyr97, His99, Glu110 and Arg121 in beta-strands H, I and J. Thus, the conformational arrangement of the ligand complex suggests that the binding affinity of ILBP for bile acid molecules is based mainly on strong hydrophobic interactions inside the protein cavity. Furthermore, this binding mode explains how ILBP can transport unconjugated and conjugated bile acids. PMID- 10806392 TI - Leptin receptor-mediated regulation of cholinergic neurotransmitter phenotype in cells of central nervous system origin. AB - Leptin is an adipocyte-secreted hormone that regulates body weight and exerts effects on hematopoiesis, reproduction, and immunity. The leptin receptor (OBR) shares sequence similarity and signaling capabilities with receptors for cytokines of the ciliary neurotrophic factor (CNTF) family. Our previous finding that CNTF and leptin exert similar anti-obesity effects and activate common neuronal signaling pathways, prompted us to investigate whether leptin may share with CNTF the ability to regulate the expression of specific neuronal genes. To this end, we established a cell line, derived from the murine septal cholinergic neuronal cell line SN-56, which stably expresses OBR. In this cell line, termed SN-56/OBR, leptin induces STAT transcription factor activation and STAT-dependent reporter gene expression in a manner similar to that of CNTF. Furthermore, in SN 56/OBR cells both CNTF and leptin produce changes in neurotransmitter and neuropeptide phenotype characteristic of cholinergic neurons, such as an increase in choline acetyltransferase and vasoactive intestinal polypeptide, and a decrease in neuropeptide Y expression. SN-56/OBR cells thus constitute an interesting new model system to investigate leptin action in cells of central nervous system origin. Possible physiological implications of OBR's intrinsic ability to regulate cholinergic phenotypic markers are discussed. PMID- 10806393 TI - Polypeptide binding properties of the chaperone calreticulin. AB - Calreticulin is a highly conserved eukaryotic ubiquitious protein located mainly in the endoplasmic reticulum. Two major characteristics of calreticulin are its chaperone activity and its lectin properties, but its precise function in intracellular protein and peptide processing remains to be elucidated. We have investigated the interactions of human calreticulin with denatured ovalbumin, proteolytic digests of ovalbumin, and different available peptides by solid phase assays, size-exclusion chromatography, capillary electrophoresis, and MS. The results show that calreticulin interacts better with unfolded ovalbumin than with native ovalbumin, that calreticulin strongly binds components in proteolytic digests of denatured ovalbumin, and that calreticulin interacts strongly with certain synthetic peptides. PMID- 10806394 TI - Oxidized galectin-1 promotes axonal regeneration in peripheral nerves but does not possess lectin properties. AB - Galectin-1 has recently been identified as a factor that regulates initial axonal growth in peripheral nerves after axotomy. Although galectin-1 is a well-known beta-galactoside-binding lectin, its potential to promote axonal regeneration as a lectin has not been reported. It is essential that the process of initial repair in peripheral nerves after axotomy is well clarified. We therefore undertook to investigate the relation between the structure and axonal regeneration-promoting activity of galectin-1. Recombinant human galectin-1 secreted into the culture supernatant of transfected COS1 cells (rhGAL-1/COS1) was purified under nonreducing conditions and subjected to structural analysis. Mass spectrometric analysis of peptide fragments from rhGAL-1/COS1 revealed that the secreted protein exists as an oxidized form containing three intramolecular disulfide bonds (Cys2-Cys130, Cys16-Cys88 and Cys42-Cys60). Recombinant human galectin-1 (rhGAL-1) and a galectin-1 mutant in which all six cysteine residues were replaced by serine (CSGAL-1) were expressed in and purified from Escherichia coli for further analysis; the purified rhGAL-1 was subjected to oxidation, which induced the same pattern of disulfide linkages as that observed in rhGAL-1/COS1. Oxidized rhGAL-1 enhanced axonal regeneration from the transected nerve sites of adult rat dorsal root ganglion explants with associated nerve stumps (5.0-5000 pg. mL-1), but it lacked lectin activity. In contrast, CSGAL-1 induced hemagglutination of rabbit erythrocytes but lacked axonal regeneration-promoting activity. These results indicate that galectin-1 promotes axonal regeneration only in the oxidized form containing three intramolecular disulfide bonds, not in the reduced form which exhibits lectin activity. PMID- 10806395 TI - An evolutionarily conserved tripartite tryptophan motif stabilizes the prodomains of cathepsin L-like cysteine proteases. AB - Cathepsin L-like cysteine proteinases contain an evolutionarily highly conserved alpha-helical motif in the proregion. This is called the ER(F/W)N(I/V)N motif according to the conserved amino acids along one side of the helix. We studied the function of this motif using site-directed mutagenesis experiments of human procathepsin S. We replaced each of these amino acids with alanine and constructed deletion mutants lacking parts of the helix. All mutants were expressed in HEK 293 cells, but only one, W52A, was not processed to mature cathepsin S, nor was it phosphorylated or secreted into the culture medium. W52 is part of the hydrophobic core in the propeptide region of cathepsin S comprising two additional tryptophan residues, W28 and W31, also conserved among cathepsin L-like cysteine peptidases. Replacement of the latter with alanine led to consequences similar to those with the W52A mutation. Recombinant propeptides containing mutations of one of the three tryptophan residues were three orders of magnitude less effective as inhibitors of mature cathepsin S than the wild-type propeptide. The results point to a dominant role of the respective hydrophobic stack in the proper folding, transport and maturation of procathepsin S and related cathepsin L-like cysteine proteinases. PMID- 10806396 TI - Binding of rat brain hexokinase to recombinant yeast mitochondria: identification of necessary physico-chemical determinants. AB - The association of rat brain hexokinase with heterologous recombinant yeast mitochondria harboring human porin (Yh) is comparable to that with rat liver mitochondria in terms of cation requirements, cooperativity in binding, and the effect of amphipathic compounds. Mg2+, which is required for hexokinase binding to all mitochondria, can be replaced by other cations. The efficiency of hexokinases, however, depends on the valence of hydrophilic cations, or the partition of hydrophobic cations in the membrane, implying that these act by reducing a prohibitive negative surface charge density on the outer membrane rather than fulfilling a specific structural requirement. Macromolecular crowding (using dextran) has dual effects. Dextran added in excess increases hexokinase binding to yeast mitochondria, according to the porin molecule they harbor. This effect, significant with wild-type yeast mitochondria, is only marginal with Yh as well as rat mitochondria. On the other hand, an increase in the number of hexokinase binding sites on mitochondria is also observed. This increase, moderate in wild-type organelles, is more pronounced with Yh. Finally, dextran, which has no effect on the modulation of hexokinase binding by cations, abolishes the inhibitory effect of amphipathic compounds. Thus, while hexokinase binding to mitochondria is predetermined by the porin molecule, the organization of the latter in the membrane plays a critical role as well, indicative that porin must associate with other mitochondrial components to form competent binding sites on the outer membrane. PMID- 10806397 TI - Identification, purification and characterization of an acetoacetyl-CoA thiolase from rat liver peroxisomes. AB - Acetoacetyl-CoA specific thiolases catalyse the cleavage of acetoacetyl-CoA into two molecules of acetyl-CoA and the synthesis (reverse reaction) of acetoacetyl CoA. The formation of acetoacetyl-CoA is the first step in cholesterol and ketone body synthesis. In this report we describe the identification of a novel acetoacetyl-CoA thiolase and its purification from isolated rat liver peroxisomes by column chromatography. The enzyme, which is a homotetramer with a subunit molecular mass of 42 kDa, could be distinguished from the cytosolic and mitochondrial acetoacetyl-CoA thiolases by its chromatographic behaviour, kinetic characteristics and partial internal amino-acid sequences. The enzyme did not catalyse the cleavage of medium or long chain 3-oxoacyl-CoAs. The enzyme cross reacted with polyclonal antibodies raised against cytosolic acetoacetyl-CoA thiolase. The latter property was exploited to confirm the peroxisomal localization of the novel thiolase in subcellular fractionation experiments. The peroxisomal acetoacetyl-CoA thiolase most probably catalyses the first reaction in peroxisomal cholesterol and dolichol synthesis. In addition, its presence in peroxisomes along with the other enzymes of the ketogenic pathway indicates that the ketogenic potential of peroxisomes needs to be re-evaluated. PMID- 10806398 TI - Down-regulation of anandamide hydrolase in mouse uterus by sex hormones. AB - Endocannabinoids are an emerging class of lipid mediators, which mimic several effects of cannabinoids. Anandamide (arachidonoylethanolamide) is a major endocannabinoid, which has been shown to impair pregnancy and embryo development. The activity of anandamide is controlled by cellular uptake through a specific transporter and intracellular degradation by the enzyme anandamide hydrolase (fatty acid amide hydrolase, FAAH). We characterized FAAH in mouse uterus by radiochromatographic and immunochemical techniques, showing that the enzyme is confined to the epithelium and its activity decreases appreciably during pregnancy or pseudopregnancy because of lower gene expression at the translational level. Ovariectomy prevented the decrease in FAAH, and both progesterone and estrogen further reduced its basal levels, suggesting hormonal control of the enzyme. Anandamide was shown to induce programmed cell death in mouse blastocysts, through a pathway independent of type-1 cannabinoid receptor. Blastocysts, however, have a specific anandamide transporter and FAAH, which scavenge this lipid. Taken together, these results provide evidence of an interplay between endocannabinoids and sex hormones in pregnancy. These findings may also be relevant for human fertility, as epithelial cells from healthy human uterus showed FAAH activity and expression, which in adenocarcinoma cells was increased fivefold. PMID- 10806399 TI - The cationic C-terminus of rat Muc2 facilitates dimer formation post translationally and is subsequently removed by furin. AB - Earlier immunolocalization experiments showed that the extreme cationic C terminus of the rat intestinal mucin Muc2 (RMC) was present at the base of intestinal goblet cells in the vicinity of ER and golgi compartments, but was not found with the rest of the mucin in apical storage granules. This prompted us to investigate the possibility that an early proteolytic cleavage reaction occurs post-translationally. A plasmid pRMC, encoding the C-terminal 534 amino acids of the mucin, was expressed in COS-7 cells and was shown to undergo cleavage at an R T-R-R sequence located within the C-terminal 14 amino acids. Cleavage did not occur with the construct RMCfH, a furin site-mutated (A-T-A-A) counterpart of pRMCH (poly His6 tagged RMC). Addition of a furin inhibitor to COS-7 cell incubations also prevented cleavage of RMC and RMCH products. 35S pulse-chase kinetic experiments revealed that a truncated mutant lacking the C-terminal 14 amino acids (pRMCDeltaCT) forms faulty (doublet) dimers in the ER. These were not secreted as efficiently as the normal dimer of wild-type (pRMC) constructs. Thus the cationic C-terminus of rMuc2 apppears to facilitate the correct formation of normal Muc2 domain dimers. PMID- 10806400 TI - Cloning, overexpression and mutagenesis of cDNA encoding dihydrolipoamide succinyltransferase component of the porcine 2-oxoglutarate dehydrogenase complex. AB - Dihydrolipoamide succinyltransferase (E2o) is the structural and catalytic core of the 2-oxoglutarate dehydrogenase (OGDH) complex. The cDNA encoding porcine E2o (PE2o) has been cloned. The PE2o cDNA spans 2547 bases encoding a presequence (68 amino-acid residues) and a mature protein (387 residues, Mr = 41 534). Recombinant porcine E2o (rPE2o) (residues 1-387), C- and N-terminal truncated PE2os, and site-directed mutant PE2os were overexpressed in Escherichia coli via the expression vector pET-11d and purified. The succinyltransferase activity of the rPE2o was about 2.2-fold higher than that of the native PE2o. Electron micrographs of the rPE2o negatively stained showed a cube-like structure very similar to that of the native PE2o. Deletion of five amino-acid residues from the C-terminus resulted in a complete loss of both enzymatic activity and formation of the cube-like structure, but the deletion of only the last two residues had no effect on either function, suggesting the important roles of the C-terminal leucine triplet (Leu383-384-385). Substitution of Ser306 with Ala, and Asp362 with Asn, Glu or Ala in the putative active site, and Leu383-384-385 with Ala or Asp abolished both functions. Substitution of His358 with Cys resulted in an 8.5 fold reduction in kcat, with little change in Km values for dihydrolipoamide and succinyl-CoA. However, self-assembly was not affected. These data indicate that Ser306, Asp362 and the Leu383-384-385 triplet are important residues in both the self-assembly and catalytic mechanism of PE2o. PMID- 10806401 TI - Novel high-affinity photoactivatable antagonists of corticotropin-releasing factor (CRF) photoaffinity labeling studies on CRF receptor, type 1 (CRFR1). AB - Novel photoactivatable antagonists of human/rat corticotropin-releasing factor (h/rCRF) have been synthesized and characterized. The N-terminal amino acid D phenylalanine in astressin ?cyclo(30-33) [D-Phe12, Nle21,38, Glu30, Lys33]h/rCRF (12-41)?, a potent CRF peptide antagonist, was replaced by a phenyldiazirine, the 4-(1-azi-2,2,2-trifluoroethyl)benzoyl (ATB) residue. Additionally, His32 of astressin was substituted by either alanine or tyrosine for specific radioactive labeling with 125I at either His13 or Tyr32, respectively. The photoactivatable CRF antagonists were tested for their ability to displace 125I-labeled Tyr0 ovine CRF ([125I-labeled Tyr0]oCRF) in binding experiments and to inhibit oCRF stimulated adenylate cyclase activity in human embryonic kidney (HEK) 293 cells, permanently transfected with cDNA coding for rat CRF receptor, type 1 (rCRFR1) or human Y-79 retinoblastoma cells known to carry endogenous functional human CRFR1 (hCRFR1). ATB-cyclo(30-33)[Nle21,38, Glu30, Ala32, Lys33]h/rCRF-(13-41) (compound 1) was found to bind with higher affinity to rat or human CRFR1 when compared with ATB-cyclo(30-33)[Nle21,38, Glu30, Tyr32, Lys33]h/rCRF-(13-41) (compound 2) and exhibited higher inhibition of oCRF-stimulated cAMP accumulation in HEK 293 cells stably transfected with cDNA coding for rCRFR1 (HEK-rCRFR1 cells) or Y-79 cells. A highly glycosylated, 66-kDa protein was identified with SDS/PAGE, when the radioactively iodinated compounds 1 or 2 were covalently linked to rCRFR1. The specificity of the photoactivatable 125I-labeled CRF antagonists was demonstrated with SDS/PAGE by the finding that these analogs could be displaced from the receptor by their corresponding nonlabeled form, but not other unrelated peptides such as vasoactive intestinal peptide. The observed molecular size of the receptor was in agreement with the size of CRFR1 found in rat pituitary (66 kDa), but was significantly larger than the size of CRFR1 found in rat cerebellum and olfactory bulb (53 kDa). PMID- 10806402 TI - Detection and distribution patterns of telomerase activity in insects. AB - Telomeres of most insects consist of pentanucleotide (TTAGG)n repeats, although the repeats are absent in Diptera and some other insect species, where the telomere regions are perhaps maintained without telomerase. To understand various and unusual telomere formation in insects, we have studied the characteristic features of a putative insect telomerase that has not been previously described. Using a modified telomeric repeat amplification protocol (TRAP), we first detected the telomerase activity in crickets, cockroaches and two Lepidopteran insects. The telomerase from crickets and cockroaches required dATP, dGTP and dTTP but not dCTP as a substrate and sequence analyses of the products of TRAP revealed that the (TTAGG)n repeats are synthesized by telomerase. The cockroach telomerase was detected both in somatic (fat body, muscle and neural tissues) and germ line (testis) cells, suggesting that expression of this enzyme is not regulated in a tissue-specific manner at an adult stage. While we detected high levels of telomerase activity in crickets and cockroaches, we could not detect activity in all tissues and cell cultures of the silkworm, Bombyx mori and in two Drosophila and one Sarcophaga cell lines. This supports the theory that Dipteran insects maintain their telomeres without telomerase. PMID- 10806403 TI - Intrinsic conformation of lipid A is responsible for agonistic and antagonistic activity. AB - Lipopolysaccharides (LPS, endotoxin) represent a major virulence factor of Gram negative bacteria, which can cause septic shock in mammals, including man. The lipid anchor of LPS to the bacterial outer membrane, lipid A, exhibits a peculiar chemical structure, harbours the 'endotoxic principle' of LPS and is also responsible for the expression of pathophysiological effects. Chemically modified lipid A can be endotoxically inactive, but may express strong antagonistic activity against endotoxically active LPS. By applying orientation measurements with attenuated total reflectance (ATR) infrared spectroscopy on hydrated lipid A samples, we show here that these different biological activities are directly correlated to the intrinsic conformation of lipid A. Bisphosphoryl-hexaacyl lipid A molecules with an asymmetric (4/2) distribution of the acyl chains linked to the diglucosamine backbone have a large tilt angle (> 45 degrees ) of the diglucosamine backbone with respect to the membrane surface, a conical molecular shape (larger cross-section of the hydrophobic than the hydrophilic moiety), and are endotoxically highly active. Monophosphoryl hexaacyl lipid A has a smaller tilt angle, and the conical shape is less expressed in favour of a more cylindrical shape. This correlates with decreasing endotoxic activity. Penta- and tetraacyl lipid A or hexaacyl lipid A with a symmetric acyl chain distribution (3/3) have a small tilt angle (< 25 degrees ) and a cylindrical shape and are endotoxically inactive, but may be antagonistic. PMID- 10806404 TI - Secondary structure composition of reconstituted subunit b of the Escherichia coli ATP synthase. AB - Subunit b of the Escherichia coli ATP synthase was isolated by preparative gel electrophoresis, acetone precipitated and after ion-pair extraction redissolved in a buffer either containing n-dodecyl-beta-D-maltoside or sodium cholate. The secondary structure of isolated subunit b was shown to be the same as within the FO complex, but was strongly dependent on the detergent used for replacement of the phospholipid environment. This was shown by an identical tryptic digestion pattern, which was strongly influenced by the detergent used for solubilization. An influence of the detergent n-dodecyl-beta-D-maltoside on the secondary structure of the hydrophilic part of subunit b was also shown for the soluble part of the polypeptide comprising residues Val25 to Leu156 (bsol) using CD spectroscopy. In order to determine the secondary structure of subunit b in its native conformation, isolated subunit b was reconstituted into E. coli lipid vesicles and analyzed with CD spectroscopy. The resulting spectrum revealed a secondary structure composition of 80% alpha helix together with 14% beta turn conformation. These results suggest that subunit b is not a rigid rod-like alpha helix simply linking F1 to FO, but rather provides an inherent flexibility for the storage of elastic energy within the second stalk generated by rotational movements within the F1FO complex. PMID- 10806405 TI - Effects of aluminum on activity of krebs cycle enzymes and glutamate dehydrogenase in rat brain homogenate. AB - Aluminum is a neurotoxic agent for animals and humans that has been implicated as an etiological factor in several neurodegenerative diseases and as a destabilizer of cell membranes. Due to its high reactivity, Al3+ is able to interfere with several biological functions, including enzymatic activities in key metabolic pathways. In this paper we report that, among the enzymes that constitute the Krebs cycle, only two are activated by aluminum: alpha-ketoglutarate dehydrogenase and succinate dehydrogenase. In contrast, aconitase, shows decreased activity in the presence of the metal ion. Al3+ also inhibits glutamate dehydrogenase, an allosteric enzyme that is closely linked to the Krebs cycle. A possible correlation between aluminum, the Krebs cycle and aging processes is discussed. PMID- 10806406 TI - Probing ligand-induced conformational changes of human CD38. AB - The lymphoid surface antigen CD38 is basically a NAD+glycohydrolase, which is also involved in the metabolism of cyclic ADP-ribose. Besides, this ecto-enzyme has potential signalling roles in T- and B-cells. Such multiple functions prompted us to study the molecular dynamics of the CD38 protein and especially the relationship between its ecto-enzymatic active site and its epitope, i.e. the binding site of most known anti-CD38 monoclonal antibodies. Both epitopic and enzymatic sites were shown to be degraded by proteases, such as trypsin or chymotrypsin. This sensitivity was almost entirely suppressed in the presence of substrates or inhibitors. Both sites were also degraded in the presence of reducing agents, as dithiothreitol. Inhibitory ligands induced the same resistance of both sites against reducing attack. The binding of CD38 ligands to the active site triggers therefore conformational changes that shield some backbone bonds and disulfide bridges against, respectively, proteolytic cleavage or reduction. This transconformation was found moreover to irreversibly take place after incubation with substrates such as NAD+ in the presence of dithiothreitol. The epitope remained preserved, while the enzymatic activity was lost. This inactivation probably resulted from the covalent trapping of the catalytically reactive intermediate in the active site (i.e. paracatalytic inactivation). These data have major implications in the knowledge of the CD38 structure, especially with regard to the location of disulfide bridges and their accessibility. Potential consequences of the conformational plasticity of CD38 should also be considered in its physiological functions such as signalling. PMID- 10806407 TI - Ser/Thr phosphorylation of hematopoietic specific protein 1 (HS1): implication of protein kinase CK2. AB - Hematopoietic lineage cell-specific protein 1 (HS1), a tyrosine multiphosphorylated protein implicated in receptor-mediated apoptosis and proliferative responses, is shown here to become Ser/Thr phosphorylated upon incubation of platelets with radiolabeled inorganic phosphate. The in vivo Ser/Thr phosphorylation of HS1 is enhanced by okadaic acid and reduced by specific inhibitors of casein kinase (CK)2. In vitro, HS1 is an excellent substrate for either CK2 alpha subunit alone (Km = 47 nM) or CK2 holoenzyme, tested in the presence of polylysine (Km = 400 nM). Phosphorylation reaches a stoichiometry of about 2 mol phosphate per mol HS1 and occurs mainly at threonyl residue(s), mostly located in the N-terminal region, but also at seryl residue(s) residing in the central core of the molecule (208-402), as judged from experiments with deleted forms of HS1. Ser/Thr phosphorylation of HS1, either induced in vivo by okadaic acid or catalysed in vitro by CK2, potentiates subsequent phosphorylation at tyrosyl residues. These data indicate the possibility that regulation of HS1 may also be under the control of Ser/Thr phosphorylation, and suggest that in quiescent cells CK2 could play a role in inducing constitutive Tyr phosphorylation of HS1 in the absence of stimuli that activate the protein tyrosine kinase pathway. PMID- 10806408 TI - Germ cell-specific heat shock protein 105 binds to p53 in a temperature-sensitive manner in rat testis. AB - Heat shock protein (HSP)105 is a testis-specific and HSP90-related protein. The aim of this study was to explore the functions of HSP105 in the rat testis. Signals of HSP105 were detected immunohistochemically in the germ cells and translocated from the cytoplasm to the nucleus at 2 days after experimental induction of cryptorchidism. In cultured testicular germ cells, a significant increase in the expression of HSP105 in response to heat stress (37 degrees C) was detected in the insoluble protein fractions. Several binding proteins were isolated from rat testis using a HSP105 antibody immunoaffinity column, and p53, the tumor suppressor gene product, was copurified with these. Furthermore, immunoprecipitation using antibodies to p53 led to coprecipitation of HSP105 together with p53 after culturing germ cells at 32.5 degrees C, but not at 37 or 42 degrees C. In conclusion, HSP105 is specifically localized in the germ cells and may translocate into the nucleus after heat shock. HSP105 is suggested to form a complex with p53 at the scrotal temperature, and dissociate from it at suprascrotal temperatures. At scrotal temperature, HSP105 may thus contribute to the stabilization of p53 proteins in the cytoplasm of the germ cells, preventing the potential induction of apoptosis by p53. PMID- 10806409 TI - Ligand-binding properties and structural characterization of a novel rat odorant binding protein variant. AB - After characterization of a novel odorant-binding protein (OBP) variant isolated from the rat nasal mucus, the corresponding cDNA was cloned by RT-PCR. Recombinant OBP-1F, the sequence of which is close to that of previously reported rat OBP-1, has been secreted by the yeast Pichia pastoris at a concentration of 80 mg.L-1 in a form identical to the natural protein as shown by MS, N-terminal sequencing and CD. We observed that, in contrast with porcine OBP-1, purified recombinant OBP-1F is a homodimer exhibiting two disulfide bonds (C44-C48 and C63 C155), a pairing close to that of hamster aphrodisin. OBP-1F interacts with fluorescent probe 1-aminoanthracene (1-AMA) with a dissociation constant of 0.6 +/- 0. 3 microM. Fluorescence experiments revealed that 1-AMA was displaced efficiently by molecules including usual solvents such as EtOH and dimethylsulfoxide. Owing to the large OBP-1F amounts expressed, we set up a novel biomimetic assay (volatile-odorant binding assay) to study the uptake of airborne odorants without radiolabelling and attempted to understand the odorant capture by OBP in the nasal mucus under natural conditions. The assay permitted observations on the binding of airborne odorants of different chemical structures and odors (2-isobutyl-3-methoxypyrazine, linalool, isoamyl acetate, 1-octanal, 1 octanol, dimethyl disulfide and methyl thiobutyrate). Uptake of airborne odorants in nearly physiological conditions strengthens the role of OBP as volatile hydrophobic odorant carriers in the mucus of the olfactory epithelium through the aqueous barrier towards the chemo-sensory cells. PMID- 10806410 TI - Functional expression of mung bean Ca2+/H+ antiporter in yeast and its intracellular localization in the hypocotyl and tobacco cells. AB - The Ca2+-transport activity and intracellular localization of the translation product of cDNA for mung bean Ca2+/H+ antiporter (VCAX1) were examined. When the cDNA was expressed in Saccharomyces cerevisiae that lacked its own genes for vacuolar Ca2+-ATPase and the antiporter, VCAX1 complemented the active Ca2+ transporters, and the microsomal membranes from the transformant showed high activity of the Ca2+/H+ antiporter. Treatment of the vacuolar membranes with a cross-linking reagent resulted in a clear band of the dimer detected with antibody specific for VCAX1p. The antibody was also used for immunolocalization of the antiporter in fractions obtained by sucrose-density-gradient centrifugation of the microsomal fraction from mung bean. The immunostained band was detected in the vacuolar membrane fraction and the slightly heavy fractions that exhibited activity of the Golgi marker enzyme. A fusion protein of VCAX1p and green fluorescent protein was expressed in tobacco cells. The green fluorescence was clearly observed on the vacuolar membrane and, in some cases, in the small vesicles. The subcellular fractionation of transformed tobacco cells confirmed the vacuolar membrane localization of the fusion protein. These results confirm that VCAX1p functions in the vacuolar membrane as a Ca2+/H+ antiporter and also suggest that VCAX1p may exist in the Golgi apparatus. PMID- 10806411 TI - Activity of environmentally relevant low doses of endocrine disruptors and the bisphenol A controversy: initial results confirmed. PMID- 10806412 TI - Reproductive malformation of the male offspring following maternal exposure to estrogenic chemicals. AB - Recently, significant concerns have been placed on the widespread use of chemicals with persistent estrogenic activity for their long-term effects on human health. In this communication, we investigated whether fetal exposure to some of these chemicals at doses consumed by people, has any long-term effect on the reproductive functions of the male offspring. Thus, time-pregnant CD-1 mice were fed diethylstilbestrol (DES), bisphenol A (BPA), and aroclor (aroclor 1016) at an average concentration of 100 ng/kg/day, 50 microg/kg/day, and 50 microg/kg/day, respectively, during Days 16-18 of gestation. A high dose of DES (200 microg/kg/day) was also tested to compare the results of the current study with those of others using the high dose only. The offspring were examined at Day 3, Day 21, and Day 60 following birth. We demonstrated that BPA, aroclor, and the lower dose of DES enhanced anogenital distance, increased prostate size, and decreased epididymal weight. No effect was found on the testicular weight or size. The chemicals also permanently increased androgen receptor (AR) binding activity of the prostate at this dosage. This is the first demonstration that environmental chemicals program AR function permanently at the dosage consumed by the general population. The higher dosage of DES, on the other hand, produced an opposite effect, decreasing prostate weight, prostate AR binding, and anogenital distance, thus confirming the previous reports. To investigate whether the above mentioned effects of the chemicals represent direct or indirect effects, we also tested the effect of the chemicals on prostate development in vitro. Thus fetal urogenital sinus (UGS), isolated at the 17th day of gestation was cultured with the chemicals in the presence and absence of testosterone (10 ng/ml) for 6 days, and prostate growth was monitored by determining the size and branching of the specimen following histology. Results showed that these chemicals induced prostate growth in the presence and absence of testosterone. They also increased androgen-binding activity. Thus, the results of the in vivo studies were reproduced in the in vitro experiments, suggesting a direct effect of these chemicals on the development of fetal reproductive organs. This is the first demonstration that estrogenic chemicals induce reproductive malformation by direct interference with the fetal reproductive organs and not by interfering with the maternal or fetal endocrine system. The chemicals are able to induce malformation even in the absence of fetal testosterone; however, they are more effective in the presence of testosterone. PMID- 10806413 TI - Water-soluble Hexasulfobutyl[60]fullerene inhibit low-density lipoprotein oxidation in aqueous and lipophilic phases. AB - Oxidative modification of low-density lipoprotein (LDL) plays a pivotal role in the pathogenesis of atherosclerosis. Increasing the resistance of LDL to oxidation may therefore mitigate, or even prevent, atherosclerosis. A new water soluble C60 derivative, hexasulfobutyl[60]fullerene [C60 - (CH2CH2CH2CH2-SO3Na)6; FC4S], consisting of 6 sulfobutyl moieties covalently bound onto the C60 cage is a potent free radical scavenger. This study explored the antioxidative effect of sulfobutylated fullerene derivatives (FC4S) on LDL oxidation. FC4S was found to be effective in protecting LDL against oxidation induced by either Cu2+ or azo peroxyl radicals generated initially in the aqueous or lipophilic phase, respectively. Levels of the oxidative products, conjugated diene and thiobarbituric acid-reactive substances, and the relative electrophoresis mobility of the LDL were decreased. The addition of 20 microM FC4S at the early stage of oxidation increased the kinetic lag time from 69 +/- 11 to 14 +/- 10 min (P < 0.05) and decreased the propagation rate from 17.1 +/- 2.6 to 6.3 +/- 1.0 mOD/min (P < 0. 005). Persistent suppression of peroxidation reaction was observed upon further addition of FC4S after full consumption of all endogenous antioxidants during the propagation period. Intravenous injection of hypercholesterolemic rabbits with FC4S (1 mg/kg/day) efficiently decreased atheroma formation. Data substantiate the use of FC4S as an excellent hydrophilic antioxidant in protecting atheroma formation, via removing free radicals, in either aqueous or lipophilic phase. PMID- 10806414 TI - Glucocorticoids maintain the extracellular matrix of differentiated mammary tissue during explant and whole organ culture. AB - Mouse mammary whole organ culture (WOC) and explant culture of lactating tissue were used to investigate the mechanism by which glucocorticoids maintain secretory epithelium following lobuloalveolar development. The relative number of mammary epithelial cells expressing glucocorticoid receptors did not change with the loss of secretory epithelium during involution as demonstrated with competitive binding assays and immunohistochemistry for the glucocorticoid receptor. Furthermore, glucocorticoids did not inhibit AP-1 binding activity. However, Northern analysis demonstrated that genes associated with the breakdown of the extracellular matrix were not expressed in tissues cultured with glucocorticoids, in contrast to their upregulation during involution of mammary tissue cultured with insulin alone. Tissue inhibitor of metalloproteinase-1 (TIMP 1) mRNA expression was lowest in tissue cultured in the presence of glucocorticoids and increased 2.3-, 3.4-, and 9-fold when tissues were involuted in the presence of insulin (Ins) alone, Ins and hydrocortisone (Hyd) with 0. 005 mg/ml, or 0.01 mg/ml collagenase IV, respectively. These data indicate that glucocorticoids maintain mammary differentiation in part by inhibiting the turnover of basement membrane. PMID- 10806415 TI - The similarity of FSH-releasing factor to lamprey gonadotropin-releasing hormone III (l-GnRH-III). AB - To validate further the existence of a specific hypothalamic follicle stimulating hormone releasing factor (FSHRF), stalk-median eminence (SME) fragments from sheep and whole hypothalami from male rats were purified by gel filtration on Sephadex G-25, and the gonadotropin-releasing activity on hemipituitaries of rats incubated in vitro was determined by bioassay and compared with the radioimmunoassayable luteinizing hormone releasing hormone (LHRH) and lamprey gonadotropin releasing hormone (l-GnRH) activities in the fractions. The FSH releasing fractions eluted in the same sequence of tubes from the Sephadex column found earlier by in vivo bioassay and were clearly separated from the immunoassayable and bioassayable LHRH. The radioimmunoassay (RIA) for l-GnRH recognized equally l-GnRH-I and -III but had negligible cross-reactivity with LHRH. Fractionation of rat hypothalamic extract by gel filtration on Sephadex G 25 revealed three peaks of l-GnRH determined by RIA, all of which eluted prior to the peak of LHRH. Only the second peak had FSH-releasing but not LH-releasing activity. To determine if this FSH-releasing activity was caused by the presence of l-GnRH in the fraction, the pituitaries were incubated with normal rabbit serum or the l-GnRH antiserum (1:1000), and the effect on the FSH- and LH releasing activity of the FSH-releasing fraction and the LH-releasing activity of LHRH was determined. The antiserum had no effect on basal release of either FSH or LH but eliminated the FSH-releasing activity of the active fraction without altering the LH-releasing activity of LHRH. Since l-GnRH-I has little activity to release FSH or LH, and its activity is nonselective, whereas previous experiments have shown that l-GnRH-III highly selectively releases FSH with a potency equal to that of LHRH to release LH, the results support the hypothesis that the FSH releasing activity observed in these experiments was caused by l-GnRH-III or a closely related peptide. PMID- 10806416 TI - Morphine tolerance in mice changes response of heroin from mu to delta opioid receptors. AB - Heroin produced antinociception in the tail flick test through mu receptors in the brain of ICR and CD-1 mice, a response inhibited by 3-O-methylnaltrexone. Tolerance to morphine was produced by subcutaneous morphine pellet implantation. By the third day, the heroin response was produced through delta opioid receptors. The response was inhibited by simultaneous intracerebroventricular (i.c. v.) administration of naltrindole, a delta opioid receptor antagonist. More specifically, delta1 rather than delta2 receptors were involved because 7 benzylidenenaltrexone, a delta1 receptor antagonist, inhibited but naltriben, a delta2 antagonist, did not. Also, antinociception produced by i.c.v. heroin was inhibited by intrathecal administration of bicuculline and picrotoxin consistent with the concept that delta1 receptors in the brain mediated the antinociceptive response through descending neuronal pathways to the spinal cord to activate GABAA and GABAB receptors rather than spinal alpha2-adrenergic and serotonergic receptors activated originally by the mu agonist action in naive mice. The mu response of 6-monoacetylmorphine, a metabolite of heroin, was changed by morphine pellet implantation to a delta2 response (inhibited by naltriben but not 7 benzylidenenaltrexone). The agonist action of morphine in these morphine-tolerant mice remained mu. Thus, the opioid receptor selectivity of heroin and 6 monoacetylmorphine in the brain is changed by production of tolerance to morphine. Such a change explains how morphine tolerant mice are not cross tolerant to heroin. PMID- 10806417 TI - Mitochondrial cytochrome c release and caspase-3-like protease activation during indomethacin-induced apoptosis in rat gastric mucosal cells. AB - Indomethacin (IND), a nonsteroidal anti-inflammatory drug, has been known to cause gastric mucosal injury as a side effect. Using a rat gastric mucosal cell line, RGM1, we determined whether apoptosis is involved in IND-mediated gastropathy, and whether caspase activation and mitochondrial cytochrome c release play an important role in producing apoptosis of IND-treated RGM1 cells in the presence of serum. IND caused caspase-3-like protease activation followed by apoptosis in a dose- and time-dependent manner. Caspase-1-like protease activity did not change during IND-induced apoptosis. IND also increased mitochondrial cytochrome c release in a time-dependent fashion. Mitochondrial cytochrome c efflux occurred just before or at the same time as caspase-3-like protease activation, and preceded the increase in apoptotic cell numbers. Z-VAD FMK, a caspase inhibitor, inhibited both the increase in caspase-3-like protease activity and apoptosis in IND-treated RGM1 cells but did not affect caspase-1 like protease activity or mitochondrial cytochrome c release. These observations suggest that the apoptosis of gastric mucosal cells could be involved in IND induced gastropathy, that cytochrome c is released from mitochondria into the cytosol during the early phase of IND-mediated apoptosis, and that subsequent activation of caspase-3-like protease, but not caspase-1-like protease, is required for the execution of apoptosis. PMID- 10806418 TI - State-dependent expression of pressure diuresis in conscious rats. AB - In 1967, Guyton and Coleman modeled pressure diuresis as the underlying, essential, long-term mechanism that regulates arterial pressure when sodium intake changes. Other mechanisms that influence renal function interact with pressure diuresis to achieve sodium balance and determine the blood pressure. Increases in sodium intake suppress sodium conserving mechanisms and activate natriuretic mechanisms; decreases in sodium intake have the opposite effect. If the Guyton-Coleman model is correct, then pressure diuresis should be more readily detected in animals on a high-salt diet than in animals on a low-salt diet. We measured spontaneous changes in arterial pressure and urine flow in conscious rats fed low-salt (0. 4% NaCl) and high-salt (8.0% NaCl) chow. For 10 rats fed a high-salt diet, arterial pressure and urine flow were positively correlated in 19 of 32 (59%) trials. In 10 rats fed a low-salt diet, a positive correlation was observed in 10 of 33 (30%) trials. Chi-square analysis revealed that differences in Na+ content of the diet were significantly associated with the probability of a positive relationship between blood pressure and urine flow. These results support the hypothesis that the expression of pressure diuresis across time is dependent on the state of sodium balance. PMID- 10806419 TI - Prevention of N-methylnitrosourea-induced colon carcinogenesis in rats by oxygenated carotenoid capsanthin and capsanthin-rich paprika juice. AB - Epidemiological and animal studies have provided evidence that dietary carotenoids may reduce the risk of certain types of cancer. An inhibitory activity of oxygenated carotenoid capsanthin, a potent antioxidant, and paprika juice rich in capsanthin (3.54 mg/100 ml) against colon carcinogenesis was investigated in F344 rats. In Experiment I (short-term assay), six rats each were given a gavage of 5 mg, 0.2 mg, or 0.008 mg capsanthin six times a week for Weeks 2-6 after receiving three intrarectal doses of 4 mg N-methylnitrosourea in Week 1. The number of colonic aberrant crypt foci, preneoplastic lesions, at Week 6 was significantly fewer (by 42%) in the 0.2 mg capsanthin group, but not in other groups, than the control group. In Experiment II (long-term assay), five groups of 30 or 25 rats each received an intrarectal dose of 2 mg N-methylnitrosourea three times a week for Weeks 1-3, and had either of 10 p.p.m. or 2 p.p.m. capsanthin solutions, 1:2.5 and 1:16.7 diluted solution of paprika juice (containing 10 p.p.m. or 2 p.p.m. capsanthin), and tap water (control fluid) as drinking fluid throughout the experiment. The experimental groups were fed 0.2 mg or 0.04 mg capsanthin/day/rat. The colon cancer incidence at Week 30 was significantly lower in the highly diluted paprika juice group (40%), but not in the moderately diluted paprika juice group (60%) and the capsanthin solution groups (68% and 68%) than the control group (83%). The results suggested that paprika juice may affect colon carcinogenesis. However, capsanthin alone failed to inhibit colon tumorigenesis, in spite of suppression of aberrant crypt foci formation in the short-term assay. Further studies are needed to explain this discrepancy. PMID- 10806420 TI - Sequence and organization analyses of a Zygosaccharomyces rouxii DNA fragment containing the HIS3 gene. AB - The nucleotide sequence of a 3.4 kb fragment containing the HIS3 gene of the osmotolerant yeast Zygosaccharomyces rouxii has been determined. The fragment was cloned from a Z. rouxii genomic DNA library by complementation of a Saccharomyces cerevisiae his3 mutant strain. The sequenced DNA fragment contained three open reading frames; the middle one (678 bp long, predicting a protein of 226 amino acids) shared a high degree of similarity with HIS3 genes of other yeast species. In the promoter region of the putative ZrHIS3 gene, a T(c) element required for constitutive transcription was found. The GenBank Accession No. of the sequenced DNA region is Y18561. PMID- 10806421 TI - Isolation and characterization of the TIM10 homologue from the yeast Pichia sorbitophila: a putative component of the mitochondrial protein import system. AB - The Saccharomyces cerevisiae TIM10 gene encodes one of the few essential mitochondrial proteins that are required for the import of nuclear-encoded precursor proteins from the cytosol and their subsequent sorting into the different mitochondrial compartments. We have isolated and characterized a putative homologue of TIM10 from the halotolerant yeast Pichia sorbitophila. The Pichia TIM10 gene encodes a protein of 90 amino acids with 66% identity to S. cerevisiae Tim10p. It was capable of suppressing the temperature sensitivity of tim10-1 mutant in S. cerevisiae, suggesting that Pichia TIM10 is both a functional and structural homologue of S. cerevisiae TIM10. The putative Pichia TIM10 gene product contains all the four conserved cysteine residues and the two CX(3)C motifs typical of the Tim family proteins in the mitochondrial intermembrane space. Using anti-Tim10p serum, Western blots detected a protein of about 10 kDa, suggesting that the Pichia Tim10p is a mitochondrial protein. The results suggest that mitochondrial import and sorting systems might be also strongly conserved in other fungi. The coding sequence of the P. sorbitophila TIM10 has been deposited in the EMBL Nucleotide Sequence Database under Accession No. AJ243940. PMID- 10806422 TI - Multiple pathways regulating fission yeast mitosis upon environmental stresses. AB - Environmental signals, such as nutrient availability and physiological stresses, modulate the cell cycle and cell size of the fission yeast Schizosaccharomyces pombe. However, little is known about how these signals are transmitted to the central cell cycle regulator, Cdc2, the cyclin-dependent kinase that induces mitosis. We show here genetic evidence that medium alkalization stimulates mitosis and consequently reduces cell size, either through the Nim1-Wee1 cascade, which regulates the inhibitory phosphorylation of Cdc2 at Tyr(15), or through the Cdc2-activating phosphatase, Cdc25. Alkaline stress stimulates phosphorylation of Nim1, accumulation of Cdc25 and dephosphorylation of Cdc2 at Tyr(15). We also show that osmostress stimulates mitosis through two independent pathways: one stimulates accumulation of Cdc25, and another dephosphorylation of Cdc2 at Tyr(15). However, our analysis demonstrates that these environmental stresses can stimulate mitosis independently of dephosphorylation of Cdc2 at Tyr(15). The S. pombe MAP kinase, Spc1, was required for the steady-state level of Cdc25 in the normal cell cycle and for its accumulation in response to alkaline stress and nutritional starvation. PMID- 10806423 TI - Regulation of pyruvate metabolism in chemostat cultures of Kluyveromyces lactis CBS 2359. AB - Regulation of currently identified genes involved in pyruvate metabolism of Kluyveromyces lactis strain CBS 2359 was studied in glucose-limited, ethanol limited and acetate-limited chemostat cultures and during a glucose pulse added to a glucose-limited steady-state culture. Enzyme activity levels of the pyruvate dehydrogenase complex, pyruvate decarboxylase, alcohol dehydrogenase, acetyl-CoA synthetase and glucose-6-phosphate dehydrogenase were determined in all steady state cultures. In addition, the mRNA levels of KlADH1-4, KlACS1, KlACS2, KlPDA1, KlPDC1 and RAG1 were monitored under steady-state conditions and during glucose pulses. In K. lactis, as in Saccharomyces cerevisiae, enzymes involved in glucose utilization (glucose-6-phosphate dehydrogenase, pyruvate dehydrogenase, pyruvate decarboxylase) showed the highest expression levels on glucose, whereas enzymes required for ethanol or acetate consumption (alcohol dehydrogenase, acetyl-CoA synthetase) showed the highest enzyme activities on ethanol. In cases where mRNA levels were determined, these corresponded well with the corresponding enzyme activities, suggesting that regulation is mostly achieved at the transcriptional level. Surprisingly, the activity of the K. lactis pyruvate dehydrogenase complex appeared to be regulated at the level of KlPDA1 transcription. The conclusions from the steady-state cultures were corroborated by glucose pulse experiments. Overall, expression of the enzymes of pyruvate metabolism in the Crabtree negative yeast K. lactis appeared to be regulated in the same way as in Crabtree positive S. cerevisiae, with one notable exception: the PDA1 gene encoding the E1alpha subunit of the pyruvate dehydrogenase complex is expressed constitutively in S. cerevisiae. PMID- 10806424 TI - Disruption of six novel ORFs from Saccharomyces cerevisiae chromosome VII and phenotypic analysis of the deletants. AB - We describe the disruption and basic phenotypic analysis of six open reading frames (ORFs) of unknown function located in the left arm of Saccharomyces cerevisiae chromosome VII, namely YGL133w, YGL134w, YGL136c, YGL138c, YGL142c and YGL144c. Disruptions were made using the short flanking homology PCR replacement strategy in the FY1679 and CEN.PK2 diploid strains. Sporulation and tetrad analysis of the heterozygous deletants was performed, as well as phenotypic analysis of the corresponding deleted haploid strains. No obvious phenotypes could be attributed to the strains deleted in any of the genes YGL134w, YGL138c and YGL144c under the conditions tested. YGL142c was shown to be an essential gene. Segregants bearing a deletion in YGL136c grew slowly in complete glycerol medium at 37 degrees C. Cells deleted in YGL133w showed abnormal morphology and reduced mating efficiency, but these phenotypes were observed only when the YGL133w disruption was in a MATalpha background. Ygl133 protein was found to localize to the nucleus. PMID- 10806425 TI - An RNA-binding protein homologue that promotes sporulation-specific gene expression in Saccharomyces cerevisiae. AB - Prior studies have shown that S. cerevisiae rim4 mutations cause reduced expression of a sporulation-specific reporter gene and sporulation. We report here that RIM4 (ORF YHL024W) is a putative RNA-binding protein of the RRM class that is expressed at elevated levels early in meiosis. Mutations in the two RRMs reduce or abolish sporulation and, in some cases, cause reduced Rim4p expression. RIM4 is required for expression of early and middle sporulation-specific genes. Unlike other meiotic regulatory genes, RIM4 is required for full gene activation through both the Ime1p and Ime2p pathways. The rim4Delta defect in activation by Ime2p is suppressed by a hyperactive Ime2p derivative. These observations argue that Rim4p may act upstream of Ime2p, perhaps in a nutritional sensing pathway. PMID- 10806426 TI - The phosphatidylcholine to phosphatidylethanolamine ratio of Saccharomyces cerevisiae varies with the growth phase. AB - This study compares the effect of the growth phase on the phospholipid composition and the activity of several phospholipid biosynthetic enzymes in a wild-type yeast grown in fermentable (glucose) and non-fermentable (lactate) semi synthetic and complete synthetic media. Several distinct differences as well as similarities were found. The cellular phosphatidylcholine: phosphatidylethanolamine (PC:PE) ratio was found to vary with the growth phase, with increases in PC levels at the expense of PE during the transition to stationary phase. The variation was most pronounced in semi-synthetic lactate medium, which is routinely used for the isolation of mitochondria, where the PC:PE ratio changed from 0.9 to 2.2 during this transition. Similar growth phase dependent changes in PC and PE content were observed in isolated organelles such as mitochondria, mitochondria-associated membranes and microsomes. Phosphatidylinositol (PI) levels were much higher in cells grown on lactate compared to cells grown on glucose (20% vs. 5-10%). Irrespective of the medium, PI levels increased upon entering stationary phase. The activities of the phospholipid biosynthetic enzymes phosphatidylserine synthase and the phospholipid-N-methyltransferases were found to be maximal at the end of logarithmic growth and to decrease upon entering stationary phase in all media. Cells grown on lactate displayed a significantly higher phospholipid:protein ratio than cells grown on glucose. The results are discussed in terms of regulation of phospholipid biosynthesis and membrane biogenesis in response to growth phase and carbon source. PMID- 10806427 TI - The CUP1 promoter of Saccharomyces cerevisiae is inducible by copper in Pichia pastoris. AB - We report the construction of a Pichia pastoris integrating vector which contains the inducible CUP1 promoter from Saccharomyces cerevisiae. We show that the promoter is indeed inducible by copper when used in P. pastoris and that the level of induction is dependent on the amount of copper in the medium. PMID- 10806428 TI - Cloning and characterization of the lactate-specific inducible gene KlCYB2, encoding the cytochrome b(2) of Kluyveromyces lactis. AB - In yeast the utilization of lactate requires two enzymes, the D and L-lactate ferricytochrome c oxidoreductase (D and L-LCR), which stereospecifically oxidize D- and L-lactate to pyruvate. These enzymes are nuclearly encoded and localized in mitochondria. In the yeast Kluyveromyces lactis, a mutant devoid of D- and L LCR activities and unable to grow on racemic lactate was isolated. Transformation of the mutant with a K. lactis genomic library allowed the isolation of the KlCYB2 gene, restoring the growth on lactate and the L-LCR activity. The KlCYB2 gene and its flanking regions were sequenced (Accession No. AJ243324; EMBL/GenBank databases). The deduced amino acid sequence is highly homologous to the corresponding Saccharomyces cerevisiae and Hansenula anomala protein sequences previously characterized. The homology is missed in the N-terminal region, corresponding to the presequence cleaved during import into mitochondria. Analysis of KlCYB2 gene expression indicated that, in contrast to S. cerevisiae, the major regulatory feature is induction by lactate. PMID- 10806429 TI - Current awareness on yeast. AB - In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals - search completed 16th Feb 2000) PMID- 10806430 TI - The effects of various definitions of dipping and daytime and night-time on the characterization of 24 h profiles of blood pressure. AB - BACKGROUND: Ambulatory blood pressure monitoring allows characterization of the patient's average blood pressure load as well as various profiles of blood pressure. Results from clinical studies suggest that dippers (patients whose blood pressures decrease during night-time) have a lower risk of cardiovascular events than do nondippers with similar daytime blood pressures. However, the definitions of dipping as well as of daytime and night-time vary among clinical studies. OBJECTIVE: To determine whether various definitions lead to markedly different classifications of dipper status. DESIGN AND METHODS: We analyzed 894 24 h ambulatory blood pressure recordings that had been performed at our institution according to three previously published definitions of daytime and night-time and five definitions of dipping. RESULTS: There were small but statistically significant differences among the mean daytime and night-time blood pressures determined using the various definitions. Likewise, the proportions of dippers varied significantly with definitions of dipping and of daytime and night time. CONCLUSIONS: Differences among definitions of dipping as well as of daytime and night-time lead to significant variations in the characterization of patients' 24 h blood blood pressure profiles. The results of this analysis can be used when comparing the results of clinical studies as well as in their design. PMID- 10806431 TI - In-vivo impaired T helper 1 cell development in submandibular lymph nodes due to IL-12 deficiency following antigen injection into the anterior chamber of the eye. AB - PURPOSE: To determine the functional properties of antigen-specific T cells that accumulate in lymph nodes following injection of antigen into the anterior chamber (AC). METHODS: Ovalbumin (OVA)-specific, CD4(+) transgenic T cell receptor (Tcr) T cells (KJ-126(+)) from DO11.10 mice were adoptively transferred into unirradiated syngeneic BALB/c mice who then received an injection of OVA in the AC or subcutaneously (SC). Three days later, KJ126(+) T cells in draining and non-draining lymph nodes were analyzed for clonal expansion, activation, and cell cycle status by flow cytometry. In addition, the cells' functional phenotype was assessed in vitro by proliferation assays and cytokine production by ELISA. RESULTS: Tcr T cells localized exclusively to draining lymph nodes after injection of OVA into the AC (neck) and SC (neck, axilla, brachial). In both cases, the KJ126(+) T cells displayed evidence of in-vivo activation and proliferation. T cells from AC-injected donors when stimulated in vitro with OVA produced small amounts of IL-2, but no IFN-gamma, IL-4, IL-5, IL-10, or TGFbeta. By contrast, T cells from SC-injected animals displayed a Th1-like phenotype (IL 2, IFN-gamma). The draining lymph nodes of mice that received systemic or intraocular administration of exogenous IL-12 at the time of the AC injection of OVA also contained KJ126(+) T cells that secreted IFN-gamma and IL-2. CONCLUSION: Antigen attracts antigen-specific T cells to lymph nodes that drain the injection site. Whereas responding T cells in nodes draining SC-injection sites differentiated into Th1-like cells, T cells in nodes draining AC injection failed to differentiated beyond the capacity to secrete IL-2 in response to antigen. Since the failure was reversed by exogenous IL-12, we propose that orderly differentiation of antigen-specific T cells down the Th1 pathway is aborted by the eye, because of a deficit in IL-12 production. PMID- 10806432 TI - Retinal pigment epithelial hypersensitivity, an association with vision loss: RPE hypersensitivity complicating paraneoplastic retinopathies. AB - PURPOSE: Paraneoplastic retinopathies are currently identified and classified according to specific anti-retina antibody reactions. These cancer-induced retinal degenerations can be accompanied by changes in the character of the retinal pigment epithelium (RPE) indicative of additional, and superimposed, pathologic activity. The antibody reactions of 11 paraneoplastic retinopathy patients were evaluated to identify those whose immunologic abnormalities included indications of RPE hypersensitivity. The pathologic significance of this anomaly was evaluated upon viable monolayers of RPE, in the presence and absence of complement. METHODS: Comparisons of antibody reactions were made using: (1) indirect immunohistochemistry on sections of rhesus monkey eyes; (2) Western blot reactions on extracts of rhesus monkey retina and in vitro-propagated rhesus RPE; and (3) an evaluation of antibody influence upon the metabolic activity of viable monolayers of rhesus RPE cells. RESULTS: Indirect immunohistochemistry on sections of monkey eyes identified antibodies reactive with RPE in four of the 11 cancer patients. The same four were found to identify a 57 kd protein component of these cells in Western blot reactions of RPE and to inhibit the metabolic activity of viable RPE cells through actions enhanced by the addition of complement. One subject from the reference comparison control group, a cancer free patient with age-related macular degeneration, also exhibited comparable antibody activity with RPE. CONCLUSIONS: RPE hypersensitivity can present as a superimposed immunologic complication to the retinal degenerations of paraneoplastic retinopathies and those unrelated to malignancies. When activated, humoral aspects of the immune response are able to impart pathologic effects that inhibit RPE homeostasis and, consequently, the essential supportive functions of these cells. Since this immunologic abnormality is not confined to cancer patients, it should be considered potentially harmful when encountered in association with any type of vision loss. PMID- 10806433 TI - Effect of anterior chamber-associated immune deviation (ACAID) on rat islet allograft rejection. AB - Anterior chamber-associated immune deviation (ACAID) is characterized by the systemic inhibition of delayed type hypersensitivity (DTH) reactions to antigens which have previously been placed into the anterior chamber of the eye. Since its initial characterization, ACAID has been elicited to a wide variety of antigens, including alloantigens, and has been shown to be due to the immune deviating effects of factors such as transforming growth factor beta (TGF-beta) in the aqueous humor on ocular antigen-presenting cells (APCs). ACAID can also be induced by injecting animals with nonocular APCs, such as peritoneal exudate cells (PECs), which have been precultured with TGF-beta and antigen in vitro. The objective of this study was to determine whether alloantigenic ACAID can be effective in preventing the rejection of rat islet allografts. The notion that islet allograft rejection can be inhibited by ACAID stems from an early study showing an ACAID-induced delay in the rejection of skin grafts. Furthermore, the immune cells mediating a DTH reaction are similar to those implicated in islet allograft rejection, suggesting that they, too, may be subject to inhibition by ACAID. Our results showed that in spite of successful ACAID induction to islet allografts, recipient rats consistently rejected their grafts. Cytotoxic T cell activity (which is not inhibited by ACAID) directed against donor alloantigens was high in these animals and may have accounted, in part, for graft failure. PMID- 10806434 TI - Intravenous immunoglobulin (IVIg) for the treatment of birdshot retinochoroidopathy. AB - Intravenous polyclonal immunoglobulin (IVIg) treatment has been successfully used in a number of autoimmune conditions. Birdshot retinochoroidopathy (BRC) is a bilateral autoimmune posterior uveitis which, in its progressive form, frequently requires immunosuppressive therapy. We report a clinical study aimed at determining the tolerance and efficiency of IVIg treatment in patients with active BRC. The study was conducted in an open manner. Eighteen patients were included. The initial visual acuity (VA) was < or =20/30 in 26 eyes, 20/25 in five eyes, and 20/20 in five eyes. IVIg was given as sole treatment at 1.6 g/kg every four weeks for six months, followed by injections of 1.2-1.6 g/kg at six to eight-week intervals. The mean follow-up was 39 months, ranging between 12 and 53 months. The results showed that the final VA of the 26 eyes with an initial VA of < or =20/30 was increased by two lines or more in 14 eyes (53.8%) and decreased in two (7.7%). Of the five eyes with an initial VA of 20/25, four had improved to 20/20 and one remained stable. Of the five eyes with an initial VA of 20/20, four remained stable and one deteriorated to 20/25. When present, macular edema was improved in half of the eyes on fluorescein angiography. Benign side effects were observed in 12 patients: moderate transient arterial hypertension (7), headache (6), eczematous lesions (6), and hyperthermia (4). The results suggest that IVIg may represent a safe alternative therapy for patients with BRC. PMID- 10806436 TI - Optic disk neovascularization in a patient with cytomegalovirus retinitis associated with renal transplantation. AB - PURPOSE: To report the development of optic nerve head neovascularization during the recovery phase of cytomegalovirus (CMV) retinitis in a renal allograft recipient. CASE REPORT: A 46-year-old male renal allograft recipient developed CMV retinitis seven months after transplantation. At the time of the diagnosis, the patient was being immunosuppressed with prednisone, cyclosporine, and azathioprine, and was treated with repeated intravitreal and intravenous ganciclovir. Six weeks after the initiation of therapy, optic disk neovascularization developed. This was confirmed by fluorescein angiography, which showed no areas of retinal capillary nonperfusion. At this stage, active retinal lesions were partially resolved. Apart from intraocular inflammation, no other cause of neovascularization was detected. Over the following six months, optic disk neovascularization regressed spontaneously without causing vitreous hemorrhage or visual loss. There was no recurrence of CMV retinitis during follow up. CONCLUSION: Optic disk neovascularization may develop in the healing phase of CMV retinitis in renal transplant recipients. PMID- 10806435 TI - Ocular findings in a measles epidemic among young adults. AB - PURPOSE: To present ocular findings in a measles epidemic outbreak among young adults. METHODS: Ophthalmic lesions in an unselected group of 61 young military personnel with measles were followed. Eighteen of the patients had symmetric keratitis in both eyes. They were treated with diclofenac sodium eyedrops qid in the right eye, while the left eye was observed untreated. The disappearance of the corneal lesions in both eyes was compared. RESULTS: Forty patients (65.6%) had measles conjunctivitis with bulbar and tarsal conjunctival hyperemia. Five (8.2%) had increased mucous secretion. Thirty-five (57.4%) had superficial punctate corneal epithelial and subepithelial lesions which stained with fluorescein. Ten patients had corneal lesions without evident conjunctival pathology. Fourteen patients had subconjunctival hemorrhages, 12 of whom had bilateral lesions mostly in the superonasal quadrant. The time to disappearance of corneal lesions was 4.5+/-3.2 days in the diclophenac-treated right eyes, and 4.1+/-3.8 days in the left eyes. We did not observe a significant difference in the healing time between the two eyes (p = 0.75). CONCLUSION: Measles did not cause major ocular complications in healthy patients. Keratitis was unresponsive to diclophenac sodium eyedrops with respect to healing time and end result. PMID- 10806437 TI - Corneal perforation due to late radiation therapy-induced corneal necrosis- clinicopathological correlation. AB - A clinicopathological correlation is made between corneal perforation and late radiation therapy-induced corneal necrosis in a male adolescent treated for orbital rhabdomyosarcoma. PMID- 10806438 TI - Pharmacological effects of pilocarpine on rabbit ciliary artery. AB - PURPOSE: A disturbance in ocular blood flow is an important factor in the pathogenesis of visual damage caused by glaucoma. We investigated the innervation of the ciliary artery, important to blood flow in the optic nerve head, and the effects of pilocarpine thereon. METHODS: Ciliary arteries were prepared from rabbit eyes and mounted in a myograph system. The effects of electrical nerve stimulation and pilocarpine on the isolated rabbit ciliary artery were investigated using isometric tension recording methods. RESULTS: Electrical field stimulation evoked contraction of the ciliary artery, which was inhibited by pretreatment with 10 microM bunazosin. Electrical field stimulation also evoked relaxation when the ciliary artery was pre-contracted by 10 microM histamine in the presence or absence of endothelium. This relaxation was inhibited by pretreatment with 10 microM N(G)-nitro-L-arginine methylester (L-NAME). Pilocarpine administration produced a dose-dependent muscle relaxation that was abolished by treatment with L-NAME, 1 aeM atropine, or denudation of the endothelium. CONCLUSIONS: Results indicate that the rabbit ciliary artery is innervated by at least two different types of nerves, adrenergic nerves, which evoke contraction and nerve fibers that cause an endothelium-independent release of nitric oxide (NO). Pilocarpine relaxation of this muscle was dependent on the endothelium and NO synthesis. PMID- 10806439 TI - Perlecan in the basement membrane of corneal epithelium serves as a site for P. aeruginosa binding. AB - PURPOSE: To determine whether binding of Pseudomonas aeruginosa (P. aeruginosa) to the scarified mouse cornea depends on interaction with proteoglycans (PGs). METHODS: Scarified corneas were treated with anti-proteoglycan monoclonal antibodies (MAbs), glycosaminoglycans (GAGs), heparinase III or chondroitin ABC lyase before inoculation with P. aeruginosa strain ATCC 19660 or PAO1. Scanning electron microscopy (SEM) was used to quantitate adherent bacteria. Frozen sections of unwounded and wounded mouse cornea, the latter treated or not treated with heparinase III were stained to spatially localize perlecan [core protein or heparan sulfate (HS) side chains]. Anti-perlecan MAb against the heparan sulfate proteoglycan core protein and succinyl wheat germ agglutinin (sWGA), a lectin which recognizes N-acetyl-glucosamine in heparan sulfate, respectively were used. RESULTS: Anti-perlecan MAb, as well as heparan sulfate, heparin and heparinase III decreased the binding of both bacterial strains to cornea, and the decrease was concentration-dependent. Fluorescence microscopic analysis of sections of mouse cornea immunostained with anti-perlecan MAb showed that perlecan was localized to the epithelial basement membrane. Scarification of the mouse cornea exposed perlecan in the basement membrane and increased bacterial binding to this site was consistent with this exposure. Lectin staining revealed that heparinase treatment removed heparan sulfate side chains of perlecan from the exposed basement membrane, and this removal was consistent with a decrease in bacterial binding. CONCLUSIONS: These studies provide evidence that perlecan, core protein and its heparan sulfate side chains serve as a binding site for Pseudomonas aeruginosa when the basement membrane of the cornea is exposed. PMID- 10806440 TI - Autologous iris pigment epithelial cell transplantation in monkey subretinal region. AB - PURPOSE: To establish autologous iris pigment epithelial (IPE) cell transplantation in monkey eyes. METHODS: Autologous IPE cells from three monkeys were obtained by peripheral iridectomy, and cell culture was performed with autologous serum. The cultured cells were labeled with DiI, and transplantation was performed by transvitreal approach to the submacular region of each monkey. Fundus examination, photography, and fluorescein angiography were performed monthly. Histochemical analysis and electron microscopy were also performed six months after transplantation. RESULTS: Auto IPE cells grew well in auto serum. No fluorescein leakage or retinal thickening was observed after submacular transplantation. We demonstrated the presence of autologous IPE cells in the region 6 months after transplantation by histologic and electron microscopic examination. Electron microscopy also demonstrated that the transplanted cells appeared to be less pigmented and to have less mitochondria than did the host retinal pigment epithelial cells. However, some transplanted cells looked as if they embrace the photoreceptor outer segments. DISCUSSION: Autologous IPE cell culture of the monkey was established using auto serum. Six months after transplantation of the cultured IPE cells, we could still observe the cells in the region. This method may have potential use in human disease that requires autologous cell transplantation to prevent host-graft rejection, and to provide an alternative substance that functions like retinal pigment epithelium. PMID- 10806441 TI - Interocular differences in optic disc topographic parameters in normal subjects. AB - PURPOSE: To test the interocular differences in optic disc topography in normal subjects by means of confocal scanning laser ophthalmoscopy. METHODS: Topographic measurements of the optic disc were evaluated by means of confocal scanning laser ophthalmoscopy (Heidelberg Retina Tomograph) in 314 eyes of 157 healthy volunteers. The examination was started randomly either with the right eye or the left eye. Differences between right and left eyes in disc area, cup area, cup volume, cup/disc area ratio, rim area, rim volume, maximum cup depth, cup shape measure, retinal nerve fiber layer thickness, and retinal nerve fiber cross section area for 360 degrees and for the temporal and nasal regions of the optic nerve head were evaluated by means of Student t-test. The same parameters were assessed in a subgroup of 80 elderly (age> 50 years) healthy subjects. Holm's sequentially rejective method was used for significance correction of multiple comparisons. RESULTS: Significant interocular differences in the average retinal nerve fiber layer thickness (p = 0.0010) and retinal nerve fiber layer cross section area (p = 0.0036) were found, with the right eye showing, on the average, lower values. The left eye showed a larger retinal nerve fiber layer thickness in 94 subjects (59.87%) and a larger retinal nerve fiber cross section area in 101 subjects (64. 33%). In the temporal optic disc area there were no statistically significant differences in topometric data (p> 0.05). In the nasal area, significant interocular differences in the retinal nerve fiber layer thickness (p = 0.0002) and retinal nerve fiber layer cross section area (p = 0.0003) were found. Similar results were found when the group of subjects older than 50 years was considered. CONCLUSIONS: This study demonstrates systematic interocular differences in optic disc topometric data. Such a finding, be it due to methodological or biological reasons, should be taken in consideration in clinical trials. PMID- 10806442 TI - Tranilast inhibits cell proliferation and collagen synthesis by rabbit corneal and Tenon's capsule fibroblasts. AB - PURPOSE: To determine whether tranilast, N-(3,4-dimethoxycinnamoyl) anthranilic acid, influences cell proliferation and collagen synthesis by rabbit Tenon's capsule fibroblasts (TFs) and corneal stromal fibroblasts (CFs). METHODS: Rabbit TFs and CFs (7000 cells/well) were cultured in F-12 nutrient mixture supplemented with 1% FBS, plus 0, 3, 30, or 300 microM tranilast, and the number of cells was counted 72 hrs later. To determine the effect of tranilast on collagen synthesis, cells at confluence were cultured in a medium containing 0, 3, 30, or 300 microM tranilast and labeled with 3H-proline, and the amount of radioactivity incorporated into collagenase-sensitive proteins was measured. RESULTS: At 300 microM, tranilast decreased the number of TFs by about 27% and the number of CFs by about 45%, but had no effect on cell viability. The same concentration of tranilast reduced TFs collagen synthesis and CFs collagen synthesis. CONCLUSIONS: Tranilast may inhibit scar formation after trabeculectomy for glaucoma and after excimer laser photorefractive keratectomy. PMID- 10806443 TI - Levels of soluble ICAM-1 and soluble IL-2R in the serum and aqueous humor of uveitis patients. AB - PURPOSE: To study the ocular and systemic behaviour of the immune activation markers soluble ICAM-1 and soluble IL-2R in patients with active uveitis. METHODS: We determined the levels of soluble ICAM-1 and soluble IL-2R in the aqueous humor and peripheral blood of 23 patients with active uveitis and 16 age and gender matched controls (who were to undergo cataract extraction) by means of ELISA enzyme immunoanalysis. RESULTS: Patients with uveitis presented higher levels of soluble ICAM-1 and soluble IL-2R in the aqueous humor and peripheral blood. Comparison of the aqueous humor and peripheral blood revealed higher levels of soluble ICAM-1 in the aqueous humor, while we found no differences for soluble IL-2R. Analysis of the clinical characteristics of the uveitis we studied showed no differences in the levels of these markers in either blood or aqueous humor. CONCLUSIONS: The finding of elevated levels of soluble IL-2R and soluble ICAM-1 in the aqueous humor and peripheral blood of patients with uveitis suggests the local and systemic participation of these markers in the pathogenesis of uveitis. PMID- 10806444 TI - Quantitative analysis of the effect of near lens addition on accommodation and myopigenesis. AB - PURPOSE: To develop a quantitative, objective, and scientific basis for understanding the effects of applying near bifocal additions (ADDs) on oculomotor control and myopia development. This is important because myopia is a major public health problem that affects 25% of the U.S. population and 75% or more in Asian countries. It is also associated with an increased risk for vision threatening conditions, such as retinal breaks and detachments, as well as glaucoma. METHODS: A comprehensive model of refractive error development was constructed based on a dual-interactive feedback model of accommodation and vergence, which represented the short-term dynamics pathway, with the addition of both genetic and environmental (defocus-induced axial growth) components in a long-term pathway. An alternating near- and far-viewing paradigm was simulated, with varying amounts of ADDs, to obtain a parametric relationship between the root mean square of accommodative error (AE) and the induced refractive error (IRE). The parametric relationship provided the crucial linkage between the long term growth pathway and the conventional short-term dynamics pathway. ADD is the simulated lens placed before the eyes only during near viewing, whereas IRE is the simulated lens fixed before the eyes that represents the optical effect of slowly progressive refractive development caused by near work. RESULTS: A V shaped functional relationship was found between AE and IRE. The left half of the curve is associated with hyperopic defocus and myopigenesis, whereas the right half is associated with myopic defocus and hyperogenesis. Introduction of an ADD shifts the V-shaped curve horizontally. Thus, an "optimal" ADD can be used to shift the minimum of the accommodative error curve to the zero IRE point, and thereby reduce or eliminate retinal defocus and its potential towards myopigenesis. On the other hand, sensitivity analysis of model parameters shows that increasing the accommodative convergence crosslink gain (AC) shifts the curve to the right and results in a tendency towards myopigenesis, which is consistent with clinical findings in progressive myopia. CONCLUSIONS: The model can be used to specify the precise ADD needed for an individual to retard or eliminate retinal defocus-induced myopic progression. If future experiments show that using the "optimal" ADD results in the greatest benefit (i.e., least myopia progression), there will be considerable worldwide public health benefit. PMID- 10806445 TI - Expression and localization of enzymes of arginine metabolism in the rat eye. AB - PURPOSE: Production of NO may be regulated by argininosuccinate synthetase and argininosuccinate lyase which recycle citrulline to arginine, and by arginase which hydrolyzes arginine to urea and ornithine. Expression of these and related enzymes in rat eye was studied. METHODS: mRNAs for the enzymes were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Localization of the enzyme proteins and mRNAs was analyzed by immunohistochemistry and in situ hybridization, respectively. RESULTS: In RT-PCR analysis, arginase II (nonhepatic type) mRNA was detected in retina and weakly in cornea, whereas arginase I (hepatic type) mRNA was not detected in any area. mRNAs for argininosuccinate synthetase, argininosuccinate lyase, ornithine aminotransferase and ornithine decarboxylase were present in all areas. In immunohistochemical analysis, arginase II was stained in cornea, epithelium of iris and ciliary process, inner part of neural retina and retinal pigment epithelium. Immunostaining of ornithine aminotransferase resembled that of arginase II. In in situ hybridization, arginase II and ornithine aminotransferase mRNAs were located much as seen with the enzyme proteins. CONCLUSIONS: These results indicate that arginine recycling activity from citrulline is present widely in ocular tissues, whereas expression of arginase II differs among the tissues. We suggest that NO production may be regulated by these enzymes in the cells where NO synthase is colocalized. Colocalization of arginase II and ornithine aminotransferase suggests a role for arginase II in collagen synthesis in the eye. PMID- 10806446 TI - Delayed graft rejection in pre-vascularised corneas after subconjunctival injection of clodronate liposomes. AB - PURPOSE: To test the effects of clodronate liposomes on graft survival and neovascularisation after transplantation in pre-vascularised recipient corneas. METHODS: Corneal neovascularisation was induced in F344 rats by injecting heat inactivated rabbit serum intrastromally. After 4 weeks F344 rats were orthotopically grafted with corneal buttons from DA rats. Directly after transplantation and on 2, 4, 6 and 8 days postoperatively clodronate liposomes were administrated subconjunctivally in one group, whereas the other group remained untreated. For 60 days grafts were observed for signs of graft rejection and neovascularisation. RESULTS: Graft survival was significantly prolonged, but not prevented in clodronate liposome treated rats compared to untreated rats ( p =.004). Also clodronate liposome administration delays growth of corneal neovascularisation after transplantation. CONCLUSIONS: Previous studies revealed that clodronate liposomes prevent corneal graft rejection and reduce neovascularisation in orthotopic corneal allotransplantation in rats. This study shows that also in pre-vascularised recipient corneas subconjunctival administration of clodronate liposomes seems to delay corneal graft rejection and reduces neovascularisation. PMID- 10806447 TI - Suppression of TGF-beta signaling in both normal conjunctival fibroblasts and pterygial body fibroblasts by amniotic membrane. AB - PURPOSE: When used as an alternative substrate following bare sclera removal of pterygium and other ocular surface diseases, amniotic membrane transplantation can reduce scarring on the reconstructed conjunctival surface. This study was carried out to determine if the amniotic membrane (AM) suppresses the expression of the TGFb signaling system in cultured normal conjunctival (HCF) and pterygial body fibroblasts (PBF). METHODS: HCF and PBF were cultured on AM and plastic wells in serum-containing and serum-free DMEM with or without TGF-beta1. Total RNA was extracted and subjected to Northern hybridization with probes of TGF beta1, b2 and b3; TGF-beta receptors (TGF- beta R) type I, II and III; a-smooth muscle actin (alpha-SM), b1-integrin, CD44, fibroblast growth factor receptor 1 (FGF-R1/ flg) and platelet-derived growth factor receptor b (PDGFR-beta); and GAPDH as a loading control. MTT assay was used for cell proliferation. RESULTS: Amniotic membrane markedly suppressed the transcript expression of TGF-beta2, b3 and all three types of TGF-beta receptors by both fibroblasts as compared to their cultures on plastic surface. In addition, expression of CD44 transcript was also markedly suppressed while that of b1 integrin, a-SM actin, and FGFR1/flg was mildly suppressed. In contrast, expression of TGF-beta1 and PDGFR-beta remained largely unchanged. The cell proliferation of HCF and PBF grown on AM was also significantly suppressed. CONCLUSIONS: Amniotic membrane matrix uniquely suppresses TGF- beta signaling in both types of fibroblasts. It may also suppress signaling via CD44, b1 integrin and FGFR1/flg. As a result, the phenotype may become less mitogenic, contractile and fibrogenic. These data support in part why amniotic membrane transplantation has an anti-scarring effect for conjunctival surface reconstruction. PMID- 10806448 TI - Comparison of grading schemes to quantitative assessments of nucleus-to cytoplasmic ratios for human bulbar conjunctival cells collected by impression cytology. AB - PURPOSE: To quantitatively assess the measured nucleus-to-cytoplasm (N/C) ratios for sets of bulbar conjunctival cells categorised by a subjective grading scheme (0-3). METHODS: Impression cytology samples were taken from the nasal bulbar conjunctiva of 37 individuals (aged 22 to 74 years) without any significant eye disease. The sheets of cells were generally stained with Giemsa, photographed and 35 mm slides prepared. The images were graded 0 to 3 according to the scheme of Nelson [Cornea 7, 71-81 (1988)] and approximately 35 contiguous cells from each image measured by planimetry from an overlay. RESULTS: Analysis of 1097 cells revealed that the N/C ratios ranged from 1.395 to 0.056 and correlated well with the grading. The average N/C ratio (range for +/- 1.96 SD) for each grade was 0.607 (0.983 to 0.531) for grade 0, 0.533 (0.759 to 0. 333) for grade 1, 0.291 (0.427 to 0.155) for grade 2, and 0.154 (0. 228 to 0.080) for grade 3. CONCLUSIONS: N/C ratios can be assessed from conjunctival impression cytology samples and appear to be sufficiently consistent to allow for a designated range of cell N/C ratios to be specified for each grade. PMID- 10806449 TI - Inhibition of selenite cataract by S-diethylsuccinyl glutathione isopropyl ester. AB - PURPOSE: Previous studies have demonstrated that glutathione derivatives can partially prevent loss of hepatic glutathione levels, in vivo, during periods of oxidative stress. Since cataracts in animal model systems and in humans are thought to be the direct result of oxidative insult, the following study tested the possibility that treatment with these glutathione analogues may affect the progression of lens opacification. METHODS: Glutathione esters were tested for their ability to inhibit the selenite-induced cataract in rats. RESULTS: The S alkyl glutathione ester Et(2)Sc-GS-iPr, but not a similar glutathione derivative S-succinyl glutathione (Sc-GS), at 0.5 mmoles/kg body weight, had anti-cataract activity in the selenite-induced cataract of rats. Analysis of lenses from treated and untreated animals demonstrated that Et(2)Sc-GS-iPr partially prevented the loss of reduced glutathione levels. CONCLUSIONS: The results demonstrate for the first time that an S-alkylated glutathione ester has anti cataractogenic potential in the selenite-treated rat, and suggest possible mechanisms involving glutathione in the prevention of this lens opacification. PMID- 10806450 TI - Inferring causality from relational data and designs: historical and contemporary lessons for research and clinical practice. AB - My address to you today is born of frustration with the growing claims, evident in the research literature and in clinical reports that happen to cross my desk, of causation made on the basis of correlational data. I recall, as a sophomore in college, being taught early in my first experimental psychology course, "you cannot infer causality from correlation." This point was made adamant in my first face-to-face critique of an experimental psychology research paper by Dr. Robert T. Brown, who pointed out, in lowering my grade, that I had inferred organic causes to behavior patterns in gerbils based solely on correlational data. This caution was reiterated in my statistics courses until it must have been indelibly stamped upon my then still somewhat plastic brain. PMID- 10806451 TI - WAIS-III Matrix Reasoning test performance in a mixed clinical sample. AB - This study examined the relationship between the Matrix Reasoning subtest (MRT) of the WAIS-III and a selected number of neuropsychological tests in a heterogeneous clinical sample of English-speaking American (n = 41), and non English-speaking immigrant (n = 14) adults. A moderate association between the Halstead Category Test and the MRT (-.58) was found in the English-speaking sample. Multiple regression analysis revealed a significant association between measures of verbal abstract reasoning and verbal fluency, and performance on the MRT. Among the immigrant sample, the MRT was also found to be significantly associated with verbal fluency task performance, as well as with the Comprehensive Test of Nonverbal Intelligence. Correlational analyses therefore suggest a strong verbal mediation element in the MRT, and that labeling it a nonverbal task may be misleading. PMID- 10806452 TI - Neurologic validity of the Wisconsin Card Sorting Test with a pediatric population. AB - This study investigated the neurologic validity of the Wisconsin Card Sorting Test (WCST) with a pediatric population; that is, the ability of the test to detect dysfunction in the frontal lobes. Fifty children with diverse etiologies of brain dysfunction were classified via EEG, MRI, or CT as having left hemisphere, right hemisphere, or bilateral frontal, extrafrontal, or multifocal/diffuse regions of brain dysfunction. Findings failed to support the hypotheses that WCST performance is more impaired in frontal lesions than extrafrontal or multifocal/diffuse lesions, or that WCST performance is more impaired in left hemisphere lesions than right. Although the WCST is not helpful in localizing cerebral area of dysfunction, it may still be a clinically useful test for examining processes that children use to solve complex problems. PMID- 10806453 TI - Impact of somatoform symptomatology on credibility of cognitive performance. AB - Although the detection of conscious fabrication (i.e., malingering) of cognitive symptoms has been the recipient of burgeoning interest within the last 10 years in the empirical and clinical neuropsychological literature, whether conversion/somatization results in similarly noncredible cognitive profiles has not been formally investigated. Two thirds (13 of 19) of subjects with cognitive complaints and 1-3/3-1 code types on the MMPI/MMPI-2 showed evidence of noncredible cognitive performance (i.e., failure on malingering tests and/or a "malingering" pattern on standard neuropsychological tests). These results suggest that symptom fabrication associated with somatization/conversion personality orientations can extend to noncredible cognitive complaints and not just the oft-described physical manifestations. PMID- 10806454 TI - Using intraindividual variability to detect malingering in cognitive performance. AB - The utility of measures for detecting malingering was evaluated using a simulation design in which half the participants were encouraged to do their best and half were asked to feign head injury. Particular attention was focused on the utility of repeated assessment (intraindividual variability) in discriminating the groups. Participants were tested on three occasions on measures commonly used to detect malingering including a specific symptom validity test (SVT). The results indicated that multiple measures of malingering obtained in single assessment (occasion one) discriminated the groups effectively. In addition, however, intraindividual variability in performance, particularly of indicators from the SVT, provided unique information beyond level of performance. The results suggest that response inconsistency across testing sessions may be a clinically useful measure for the detection of malingering. PMID- 10806455 TI - Proposed schedule of usual and customary test administration times. AB - A survey was sent to 500 members of the National Academy of Neuropsychology (NAN) in order to tally clinical test evaluation times. A return of 118 useable responses resulted in the table of "clinical times" (e.g., time required to administer, score, interpret, and report) presented in this paper. These are proposed as a practical guide for billing third-party payers. PMID- 10806456 TI - Normative comparisons for the controlled oral word association test following acute traumatic brain injury. AB - The Controlled Oral Word Association Test (COWAT) is widely used in clinical neuropsychology as a measure of verbal fluency. It is important for psychologists to realize that using the recently published normative data will result in different clinical conclusions because the updated normative sample performed better than the original normative sample. The purpose of this study was to compare the original and updated norms in a large sample of patients with acute traumatic brain injuries (N=669). The percentages of patients who scored below the 5th centile in each system varied as a function of brain injury severity. Moreover, a substantially larger number of patients scored in the impaired range according to the updated normative data. PMID- 10806457 TI - Psychometric properties of the Boston Qualitative Scoring System for the Rey Osterrieth Complex Figure. AB - Interrater and internal consistency reliabilities were computed for the Boston Qualitative Scoring System, a newly developed system for scoring the Rey Osterrieth Complex Figure. Subjects (N = 108) included males with right hemisphere stroke (n = 46), left hemisphere stroke (n = 20), dementia (n = 21), and normal controls (n = 21). Interrater reliability coefficients for all scales except asymmetry, which had a reliability coefficient of .2013, ranged from .6342 to .9919, with most in the good to excellent range. Internal consistency reliabilities ranged from .7774 to .9128, also with most in the good to excellent range. Discriminant indices of visuospatial accuracy scales found the scoring system to be useful in distinguishing between individuals based on visuospatial scores. PMID- 10806458 TI - Clinical utility of the Trail Making Test practice time. AB - The Trail Making Test (TMT) is one of the most frequently used measures in clinical neuropsychology. Data obtained from the TMT practice times were analyzed to determine their utility in predicting success and failure on the full version of the test and to allow establishment of criteria by which to judge administration or discontinuation of the full test. Results indicated that TMT practice times were useful in predicting successful completion of Part A and B of the TMT. Tables are provided which describe the classification accuracy of various TMT practice times. These tables allow clinicians to select a practice time cutoff and then use the cutoff as a heuristic to assist in the decision to administer the remainder of that particular part of the TMT or discontinue the test. A 20-s cutoff resulted in optimal prediction of successful completion (< 180 s) of TMT Part A. A cutoff of 30 s optimally predicted successful completion (< 300 s) of TMT Part B. PMID- 10806459 TI - Consequences of premature discontinuation of the developmental test of visual motor integration: can these tests be salvaged? AB - The administration of many neuropsychological tests includes discontinuation criteria that are often complex and time-consuming. As a result, it is not uncommon for an examiner to unintentionally discontinue the administration of a test prematurely. This investigation of 56 correctly scored Developmental Test of Visual-Motor Integration protocols was conducted to determine how the premature termination of this particular test significantly affects its overall score, and thus, its interpretation. These data indicate that premature discontinuation of this test results in a minimal loss of points overall, and thus, does not invalidate the interpretation of its score. PMID- 10806460 TI - Early clinical assessment of attention. AB - Fifty preschoolers participated in this study. Twenty-five preschoolers classified as ADHD were matched with 25 typically developing preschoolers, and assessed using three tests of attention (two vigilance tests, one visual-search test). Their behavior exhibited during these attention tests was also assessed. Compared to their peers, preschoolers classified as ADHD exhibited significantly more omission and commission errors on the visual attention test. On the visual search attention test, preschoolers classified as ADHD exhibited significantly more commission errors, and they took significantly longer to complete it. They did not exhibit significantly more omission or commission errors on the auditory attention test. The preschoolers classified as ADHD were also more vocal, more often off-task and out-of-seat, and they required more adult redirectives to return to task. Discussion is focused on the clinical value of developmentally appropriate attention tests and behavioral observation systems in the early clinical assessment of attention in very young children. PMID- 10806461 TI - Neuropsychological performance of Hispanic and non-Hispanic older adults: an epidemiologic survey. AB - Performance on a brief battery of neuropsychological tests was compared for 797 Hispanic and non-Hispanic white older adults (65-97 years) participating in a community-based epidemiological survey of Bernalillo County, New Mexico. Tests included measures of memory (Fuld Object Memory Evaluation), attention (Digit Span), verbal fluency (category naming), visuoconstruction (clock drawing), and psychomotor speed and cognitive flexibility (Color Trails). Statistically significant ethnic group differences were observed on all tests in analyses that also considered effects of age, education, gender, depressive symptoms, and a global measure of medical illness. Effect sizes were small for all measures except Digit Span and Color Trails. In dementia screening or other clinical cognitive assessment, separate ethnic group norms may be useful in interpreting results for these measures. Preliminary normative tables are provided for Hispanic older adults at two levels of age (65-74 years and 75-97 years) and four levels of education (0-6 years, 7-9 years, 10-12 years, and > 12 years). PMID- 10806462 TI - The Behavioral Dyscontrol Scale as a predictor of aggression against self or others in psychogeriatric inpatients. AB - The ability of the Behavioral Dyscontrol Scale (BDS) and its factors to predict aggression in psychogeriatric inpatients was investigated. Fifty-one neurologic and 33 psychiatric patients participated in the study. In the psychiatric group, Environmental Independence Factor (EIF: consisting of two go/no-go tasks) was more impaired in aggressive than in nonaggressive patients. In both groups, Fluid Intelligence Factor (FIF: consisting of alphanumeric sequencing, non-mirroring, and insight) was more impaired in aggressive than in nonaggressive patients. No differences between aggressive and nonaggressive patients were found for the Motor Programming Factor, BDS total score, or Mini-Mental State Exam. Receiver Operating Characteristic (ROC) curve using an EIF/FIF composite demonstrated high sensitivity and modest specificity for the psychiatric patients, yielding an Area Under the Curve (AUC) of 84%. Differences in the test's ability to predict aggression in the two diagnostic groups underscore the necessity for establishing unique norms for different populations. PMID- 10806463 TI - Normative clinical relationships between orientation and memory: age as an important moderator variable. AB - The present study examined the relationship between memory and orientation to time, place, and personal and general information, as moderated by age, education, and simple attentional ability. A heterogeneous sample of 312 clinical referrals was divided into four groups, according to delayed memory functioning. Patients with globally good, globally poor, poor visual, and poor auditory memory were at differential risk of being disoriented, with the globally poor memory patients having the greatest risk. Overall, poorly oriented patients were older and less educated, with worse recall of digits backward. Discriminant Function Analysis selected visual and auditory memory and age as predictors of orientation. Normative tables stratified by age and memory performance are presented. PMID- 10806464 TI - Rey-Osterrieth Complex Figure performance in adults with attention deficit hyperactivity disorder: a validation study of the Boston Qualitative Scoring System. AB - Rey-Osterrieth Complex Figure (ROCF) productions from 18 adults with Attention Deficit Hyperactivity Disorder (ADHD) were compared to 18 matched controls using the Boston Qualitative Scoring System (BQSS). ADHD adults showed impairment in measures of configural accuracy, planning, and neatness. A logistic regression model resulted in 75% sensitivity and 81% specificity in discriminating ADHD from control subjects. In contrast, there was no significant difference on the traditional ROCF 36-point score, and the sensitivity and specificity for the 36 point score were lower (68% and 71%, respectively). These findings suggest persisting executive dysfunction in adults with ADHD that can be detected in ROCF productions. Thus, the BQSS may be a useful tool contributing to the neuropsychological evaluation of adults with ADHD. PMID- 10806465 TI - An analysis of the Hooper Visual Organization Test with neurological patients. AB - An item analysis of the Hooper Visual Organization Test (VOT) was performed using the test protocols of 320 German-speaking neurological patients with a mean age of 51.3 years. All patients presented some form of cerebral pathology. The sample yielded a mean test score of 18.8 (SD = 6.2). Estimates of internal consistency of the total scale yielded acceptable results (>.88). Although the VOT was judged to be a useful instrument for assessing visuoperceptual and visuospatial organizational dysfunction, item characteristics of the test are not satisfactory. In particular, the item ranking does not correspond to the order of difficulty. Moreover, in their current form, scoring rules of a number of items seem to be arbitrary and not founded on empirical data. A revision of the VOT is proposed to improve its psychometric properties. PMID- 10806466 TI - So many norms, So little time PMID- 10806467 TI - Neuropsychological evaluation of the spanish-speaker: A reply to artiola I fortuny (1996) AB - In 1996 Artiola I Fortuny published a review of our book, Neuropsychological Evaluation of the Spanish Speaker (Ardila, Rosselli, & Puente, 1994), in this journal. We appreciate the time and energy taken to produce such an exhaustive review of our little volume. However, considering the extremely negative and personal nature of the review, we felt obliged to the scientific and professional community to provide reasoned information, which should help clarify the numerous misunderstandings in the review. PMID- 10806468 TI - Diagnostic criteria for malingered neurocognitive dysfunction: proposed standards for clinical practice and research. AB - Over the past 10 years, widespread and concerted research efforts have led to increasingly sophisticated and efficient methods and instruments for detecting exaggeration or fabrication of cognitive dysfunction. Despite these psychometric advances, the process of diagnosing malingering remains difficult and largely idiosyncratic. This article presents a proposed set of diagnostic criteria that define psychometric, behavioral, and collateral data indicative of possible, probable, and definite malingering of cognitive dysfunction, for use in clinical practice and for defining populations for clinical research. Relevant literature is reviewed, and limitations and benefits of the proposed criteria are discussed. PMID- 10806469 TI - Myths of neuropsychology: further considerations. AB - The publication of "Myths of Neuropsychology" (Dodrill, 1997) has stimulated three responses, each of which takes issue with points in the original publication. These three responses especially address Myth #4 ("Above-average performances on neuropsychological tests are expected when intellectual abilities are above average") and to a lesser degree Myth #1 ("We have a good knowledge of the constructs that our tests measure"). The present paper reviews the thoughts and new data which these colleagues have provided in response to the original paper, and it also presents additional data bearing on the issues at hand. Based on the composite of information now available, it is conceded that Myth #4 was not accurately worded in the original paper. This was a mistake, and an alternate wording for Myth #4 is offered. Other changes in the original publication do not appear to be required. In order to resolve many of the issues at hand and thereby advance our profession, research in a number of areas is needed, many of which are specifically identified in the present paper. PMID- 10806470 TI - Music assisted progressive muscle relaxation, progressive muscle relaxation, music listening, and silence: a comparison of relaxation techniques. AB - The purpose of this study was to compare the effects of music assisted progressive muscle relaxation (M + PMR), progressive muscle relaxation (PMR), music listening, and silence/suggestion on measures of anxiety and perceived relaxation. The study also examined participant responses to a posttreatment questionnaire to identify relationships between musical and nonmusical elements in relaxation techniques. Sixty university students participated in the study. Fifteen participants were randomly assigned to each treatment condition. Subjects were tested individually using the same relaxation script for M + PMR and PMR conditions. One-way analyses of covariance were computed to compare pre and posttest differences among groups. Results of the ANCOVA revealed no differences among groups for the State Trait Anxiety Inventory (STAI) or the Visual Analog Scale (VAS). Analysis of variance, however, revealed each treatment condition to be equally effective in producing significant changes in anxiety and perceived relaxation from the pre to posttest period. Additionally, mean score differences revealed decreases for all conditions with M + PMR eliciting the greatest amount of change. A content analysis of posttreatment questionnaire items revealed detailed information about each participant's relaxation experience, state of mind, and use of self-generated relaxation techniques. PMID- 10806471 TI - The effect of music amplitude on the relaxation response. AB - The purposes of this study were (a) to ascertain how 3 different volume levels of music affect the relaxation response both psychologically (preference scores and self-report) and physiologically (heart rate), (b) to determine the amplitude preference for relaxation among young adults, and (c) to compare differences in preference response between music and nonmusic majors and between the genders. One hundred forty-four college-age music and nonmusic majors were participants in this study. Subjects listened to 27 minutes of music while relaxing. The amplitude of the music was changed every 3 minutes in a randomized order so that each subject received loud (80-90 dB) medium (70-80 dB) or soft (60-70 dB) music 3 times each during the experimental period for a total of 9 amplitude changes. A sample of subjects wore a small heart rate monitor on their wrist and chest during the procedure. Simultaneously with the selected listening, they were encouraged to turn a dial on a Continuous Response Digital Interface (CRDI) indicating their amplitude preference for relaxation. Self-report information was gathered at the beginning and end of the experiment. Results of the CRDI analyses indicate that overall, subjects showed overwhelming preference for the soft music in comparison to medium or loud. Males, however, preferred the loud music more than females, and music majors preferred softer music over non-majors who preferred louder music. There were no differences attributed to amplitude level in the analysis of heart rate data. Analysis of the self report data yielded a wide variety of responses concerning their individual preferences, not always consistent with the empirical measures. Overall, there was an increase in relaxation reported over the duration of the experiment. Response differentiation to loudness levels indicates a long line of useful research not only on relaxation and stress reduction in health related fields, but also on the effects of background amplitude of music while studying, driving, and engaging in other cognitive and motor tasks. PMID- 10806472 TI - Perception of tension in music: musicians versus nonmusicians. AB - Music therapists are often highly trained musicians who deal with diverse populations, most of whom do not have formal musical training. Questions may arise regarding the issue of a therapist's ability to understand, and predict, the musical perceptions and preferences of a client when their own background is so different. The current work is a look at a series of studies using various musical stimuli and comparing responses of musicians and nonmusicians to perceived "musical tension." Subjects (N = 126) included adult musicians and nonmusicians as well as a case study of a father/daughter. All subjects listened to recordings through individual headphones and were physically isolated from other subjects to ensure individuality of responses. Subjects, whether adults or children, were given instructions telling them that they were about to hear a piece of music and that they would be using a Continuous Response Digital Interface (CRDI) dial to trace the musical tension they heard. No specific definition of musical tension was given to any of the subjects. In effect, individual subjects supplied their own definition, either knowingly or unknowingly, in the absence of a formal one. Results indicated that group perceptions of the points at which tension and its release were strongest are remarkably similar between musicians and nonmusicians (correlations ranged from r =.71 to r =.95). Within at least the western art music tradition the likelihood that perceptions of group responses to tension and release in music could be predicted is high. These data indicate that therapists, trained as musicians, might be able to predict with some accuracy the responses of their clients who are not trained musicians. PMID- 10806473 TI - The history of the undergraduate curriculum in music therapy. AB - The undergraduate curriculum determines the quality of student preparation and serves to ensure consistency in clinical practice, thus promoting acceptance of music therapy by the medical community. As members of the newly-formed American Music Therapy Association strive to integrate diverse viewpoints, a historical review of the curriculum may provide direction for future revisions. Columbia University offered the first music therapy course in 1919, yet the profession did not establish an official undergraduate curriculum until 1952. This paper chronicles the events occurring prior to 1952 which contributed to curriculum development. Preceding university degree offerings, hospitals provided music therapy training. Hospital music directors established detailed guidelines for both lecture courses and clinical work. Authorities also outlined specific qualifications for potential students, including musical ability and suitable personality traits, and discouraged students who did not meet these criteria. As university programs grew in number, notable figures such as Roy Underwood and E. Thayer Gaston influenced curricular decisions through their personal beliefs and professional activities. Exploring this chain of events may assist contemporary authorities in understanding and strengthening the educational foundation for music therapists. PMID- 10806474 TI - FAK integrates growth-factor and integrin signals to promote cell migration. AB - Here we show that cells lacking focal adhesion kinase (FAK) are refractory to motility signals from platelet-derived and epidermal growth factors (PDGF and EGF respectively), and that stable re-expression of FAK rescues these defects. FAK associates with activated PDGF- and EGF-receptor (PDGFR and EGFR) signalling complexes, and expression of the band-4.1-like domain at the FAK amino terminus is sufficient to mediate an interaction with activated EGFR. However, efficient EGF-stimulated cell migration also requires FAK to be targeted, by its carboxy terminal domain, to sites of integrin-receptor clustering. Although the kinase activity of FAK is not needed to promote PDGF- or EGF-stimulated cell motility, kinase-inactive FAK is transphosphorylated at the indispensable Src-kinase binding site, FAK Y397, after EGF stimulation of cells. Our results establish that FAK is an important receptor-proximal link between growth-factor-receptor and integrin signalling pathways. PMID- 10806475 TI - Kinesin's IAK tail domain inhibits initial microtubule-stimulated ADP release. AB - Kinesin undergoes a global folding conformational change from an extended active conformation at high ionic concentrations to a compact inhibited conformation at physiological ionic concentrations. Here we show that much of the observed ATPase activity of folded kinesin is due to contamination with proteolysis fragments that can still fold, but retain an activated ATPase function. In contrast, kinesin that contains an intact IAK-homology region exhibits pronounced inhibition of its ATPase activity (140-fold in 50 mM KCl) and weak net affinity for microtubules in the presence of ATP, resulting from selective inhibition of the release of ADP upon initial interaction with a microtubule. Subsequent processive cycling is only partially inhibited. Fusion proteins containing residues 883-937 of the kinesin alpha-chain bind tightly to microtubules; exposure of this microtubule-binding site in proteolysed species is probably responsible for their activated ATPase activities at low microtubule concentrations. PMID- 10806477 TI - The EAST protein of drosophila controls an expandable nuclear endoskeleton. AB - The high degree of structural order inside the nucleus suggests the existence of an internal nucleoskeleton. Our studies on the east gene of Drosophila, using the larval salivary gland polytene nucleus as a model, demonstrate the involvement of an extrachromosomal nuclear structure in modulating nuclear architecture. EAST, a novel ubiquitous protein, the product of the east (enhanced adult sensory threshold) locus, is localized to an extrachromosomal domain of the nucleus. Nuclear levels of EAST are increased in response to heat shock. Increase in nuclear EAST, whether caused by heat shock or by transgenic overexpression, results in the expansion of the extrachromosomal domain labelled by EAST, with a concomitant increase in the spacing between chromosomes. Moreover, EAST functions to promote the preferential accumulation of the proteins CP60 and actin in extrachromosomal regions of the nucleus. We propose that EAST mediates the assembly of an expandable nuclear endoskeleton which, through alterations of its volume, can modulate the spatial arrangement of chromosomes. PMID- 10806476 TI - Sphingosylphosphorylcholine is a ligand for ovarian cancer G-protein-coupled receptor 1. AB - Sphingosylphosphorylcholine (SPC) is a bioactive lipid that acts as an intracellular and extracellular signalling molecule in numerous biological processes. Many of the cellular actions of SPC are believed to be mediated by the activation of unidentified G-protein-coupled receptors. Here we show that SPC is a high-affinity ligand for an orphan receptor, ovarian cancer G-protein-coupled receptor 1 (OGR1). In OGR1-transfected cells, SPC binds to OGR1 with high affinity (Kd = 33.3 nM) and high specificity and transiently increases intracellular calcium. The specific binding of SPC to OGR1 also activates p42/44 mitogen-activated protein kinases (MAP kinases) and inhibits cell proliferation. In addition, SPC causes internalization of OGR1 in a structurally specific manner. PMID- 10806478 TI - Motor proteins regulate force interactions between microtubules and microfilaments in the axon. AB - It has long been known that microtubule depletion causes axons to retract in a microfilament-dependent manner, although it was not known whether these effects are the result of motor-generated forces on these cytoskeletal elements. Here we show that inhibition of the motor activity of cytoplasmic dynein causes the axon to retract in the presence of microtubules. This response is obliterated if microfilaments are depleted or if myosin motors are inhibited. We conclude that axonal retraction results from myosin-mediated forces on the microfilament array, and that these forces are counterbalanced or attenuated by dynein-mediated forces between the microfilament and microtubule arrays. PMID- 10806479 TI - The TSC1 tumour suppressor hamartin regulates cell adhesion through ERM proteins and the GTPase Rho. AB - Loss of the tumour-suppressor gene TSC1 is responsible for hamartoma development in tuberous sclerosis complex (TSC), which renders several organs susceptible to benign tumours. Hamartin, the protein encoded by TSC1, contains a coiled-coil domain and is expressed in most adult tissues, although its function is unknown. Here we show that hamartin interacts with the ezrin-radixin-moesin (ERM) family of actin-binding proteins. Inhibition of hamartin function in cells containing focal adhesions results in loss of adhesion to the cell substrate, whereas overexpression of hamartin in cells lacking focal adhesions results in activation of the small GTP-binding protein Rho, assembly of actin stress fibres and formation of focal adhesions. Interaction of endogenous hamartin with ERM-family proteins is required for activation of Rho by serum or by lysophosphatidic acid (LPA). Our data indicate that disruption of adhesion to the cell matrix through loss of hamartin may initiate the development of TSC hamartomas and that a Rho mediated signalling pathway regulating cell adhesion may constitute a rate limiting step in tumour formation. PMID- 10806480 TI - Dynamics and retention of misfolded proteins in native ER membranes. AB - When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 degrees C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes. PMID- 10806481 TI - Phospholipase C and termination of G-protein-mediated signalling in vivo. AB - In Drosophila photoreceptors, phospholipase C (PLC) and other signalling components form multiprotein structures through the PDZ scaffold protein INAD. Association between PLC and INAD is important for termination of responses to light; the underlying mechanism is, however, unclear. Here we report that the maintenance of large amounts of PLC in the signalling membranes by association with INAD facilitates response termination, and show that PLC functions as a GTPase-activating protein (GAP). The inactivation of the G protein by its target, the PLC, is crucial for reliable production of single-photon responses and for the high temporal and intensity resolution of the response to light. PMID- 10806482 TI - PDGF-C is a new protease-activated ligand for the PDGF alpha-receptor. AB - Platelet-derived growth factors (PDGFs) are important in many types of mesenchymal cell. Here we identify a new PDGF, PDGF-C, which binds to and activates the PDGF alpha-receptor. PDGF-C is activated by proteolysis and induces proliferation of fibroblasts when overexpressed in transgenic mice. In situ hybridization analysis in the murine embryonic kidney shows preferential expression of PDGF-C messenger RNA in the metanephric mesenchyme during epithelial conversion. Analysis of kidneys lacking the PDGF alpha-receptor shows selective loss of mesenchymal cells adjacent to sites of expression of PDGF-C mRNA; this is not found in kidneys from animals lacking PDGF-A or both PDGF-A and PDGF-B, indicating that PDGF-C may have a unique function. PMID- 10806484 TI - A year of Nature Cell Biology. PMID- 10806483 TI - Gli regulation by the opposing activities of fused and suppressor of fused. PMID- 10806485 TI - An encore for ribosome biogenesis in the control of cell proliferation. AB - Although the importance of cell growth for cell-cycle progression has been recognized for thirty years, the molecular basis of this relationship is poorly understood. However, researchers have begun to tease apart these two processes in model systems. This commentary focuses on one potential mechanism by which ribosome biogenesis antagonizes cell-cycle progression until the cell has grown to an adequate size. PMID- 10806486 TI - Diffusional protein transport within the nucleus: a message in the medium. PMID- 10806487 TI - The nucleoskeleton: go EAST, young man. PMID- 10806488 TI - Origins of antigenic peptides. PMID- 10806489 TI - A new twist for the tumour suppressor hamartin. PMID- 10806490 TI - A new member of an old family. PMID- 10806491 TI - Morphogen diffusion: the case of the wingless protein. PMID- 10806492 TI - It's good to talk: cell-cell communication by gap junctions. PMID- 10806493 TI - PLC fills a GAP in G-protein-coupled signalling. PMID- 10806494 TI - The transcriptional role of PML and the nuclear body. AB - The PML gene encodes a tumour suppressor protein associated with a distinct subnuclear domain, the nuclear body. Various functions have been attributed to the PML nuclear body, but its main biochemical role is still unclear. Recent findings indicate that PML is essential for the proper formation of the nuclear body and can act as a transcriptional co-factor. Here we summarize the current understanding of the biological functions of PML and the nuclear body, and discuss a role for these intra-nuclear structures in the regulation of transcription. PMID- 10806495 TI - Internal morphology of human cervical pedicles. AB - STUDY DESIGN: The internal architecture of cervical spine pedicles was investigated by thin sectioning and digitization of radiographic images. OBJECTIVES: To provide quantitative information on the internal dimensions and cortical shell thicknesses of the middle and lower cervical pedicles. SUMMARY OF BACKGROUND DATA: Although there have been a number of studies presenting data on the external dimensions of the cervical pedicle, little is known regarding its internal architecture and cortical shell thickness along the pedicle axis. METHODS: Twenty-five human cervical vertebrae (C3-C7) were secured to a thin sectioning machine to produce three 0.7-mm-thick pedicle slices along its axis. Plain radiographs of the pedicle slices were scanned and digitized to facilitate measurement of the internal dimensions. Computer software was specifically developed to determine the external dimensions (i.e., pedicle height and width) and the internal dimensions (i.e., cortical shell thicknesses of the superior, inferior, lateral, and medial walls and the cancellous core height and width) of cervical pedicles. RESULTS: Superior and inferior wall cortical thicknesses of pedicle thin slices were similar, whereas the lateral wall cortical thickness was significantly smaller than the medial wall thickness. The medial cortical shell (average value range: 1.2-2.0 mm) was measured to be 1.4 to 3.6 times as thick as the lateral cortical shell (average value range: 0.4-1.1 mm). When medial and lateral cortical thicknesses were normalized for external dimensions, the combined cortical shell thickness was thinnest at C7 (average value range: 18. 6 25.6% of the external width), and this result was statistically significant when compared with other vertebral levels. CONCLUSIONS: The cervical pedicle is a complex, three-dimensional structure exhibiting extensive variability in internal morphology. Characteristics of the cervical pedicle at different spinal levels must be noted before transpedicular screw fixation. PMID- 10806496 TI - Spinal glial activation and cytokine expression after lumbar root injury in the rat. AB - STUDY DESIGN: This study was designed to examine the behaviorial immunohistochemical changes of spinal glial cells and spinal Interleukin (IL) 1beta expression after various nerve root injuries used as models of lumbar radiculopathy. OBJECTIVES: In order to better understand the role of central inflammation in the pathophysiologic mechanisms that give rise to pain associated with lumbar radiculopathy, this research studied the relationship between pain related behavior associated with spinal glial activation and IL-1beta expression generated by three types of nerve root injury: loose ligation with chromic gut, loose ligation with silk, and tight ligation with silk. SUMMARY OF BACKGROUND DATA: An animal model of lumbar radiculopathy originally described by Kawakami and Weinstein involved loose ligation of unilateral L4-L6 nerve roots with chromic gut. Characterization and establishment of such an animal model of low back pain enables further investigation of the nature of the pathophysiologic mechanisms associated with lumbar radiculopathy in humans. METHODS: Seventy-three rats were divided into four treatment groups. Chromic group (n = 25): The L5 nerve roots (dorsal and ventral) were exposed by hemilaminectomy and loosely ligated with chromic gut. Tight silk group (n = 18): The exposed L5 nerve roots were tightly ligated extradurally with 5-0 silk suture. Loose silk group (n = 15): two loose ligatures of 5-0 silk were placed around the exposed L5 nerve roots. Sham group (n = 15): the rats were subjected to laminectomy alone for exposing nerve roots. Following surgery, thermal hyperalgesia and mechanical allodynia was assessed time-dependently up to 42 days post operatively. At 1, 3, 7, 14, and 42 days postoperatively, the rats in each group were perfused with fixative. The L5 spinal cord segments was harvested and cryosectioned for glial and cytokine immunohistochemistry. RESULTS: In the chromic and the tight silk group, an immediate and sustained mechanical allodynia was observed in the ipsilateral hind paw up to 35 days postoperatively. The loose silk group also showed an immediate mechanical allodynia that subsided by 14 days postoperatively. Sham-treated animals exhibited mild mechanicalallodynia for the initial 7 days after the surgery. Thermalhyperalgesia was evident in the three primary treatment groups, but not in the sham-treated rats. OX-42 expression was elevated in the gray matter of the L5 spinal section by 3 days in the chromic, the tight silk, and the loose silk groups as compared to the sham group. Astrocytic activation increased over time in all groups except the sham group. There was no direct correlation between degree of microglial response and severity of pain behaviors. In contrast, astrocytic activation demonstrated a direct relationship with the elevation of mechanical allodynia for the first 7 days. In addition, spinal IL-1beta protein expression was increased bilaterally in the superficial layer of the dorsal horn and cell nuclei of the ventral horns in the ligature treated groups as compared with the sham group. CONCLUSION: Direct mechanical and/or chemical injury to lumbar roots in the rat gives rise to pain behavior suggestive of lumbar radiculopathy. The finding that glial activation and enhanced IL-1beta expression are observed in the spinal cord after root injury supports a central, neuroimmune component in the generation of lumbar radiculopathy. A further understanding of the immunologic consequences of root injury may lead to further development and the novel use of selective cytokine inflammatory inhibitors for the treatment of low back pain associated with radiculopathy. PMID- 10806497 TI - Perfusion-limited recovery of evoked potential function after spinal cord injury. AB - STUDY DESIGN: The current study was designed to determine whether progressive spinal cord damage during residual compression is caused by low blood flow and ischemia. OBJECTIVES: The purpose of this experiment was to determine the effects of sustained spinal cord compression on regional blood flow and evoked potential recovery after time-dependent decompression. SUMMARY OF BACKGROUND DATA: Spinal cord injury after trauma is commonly associated with residual cord compression. Although decreased blood flow has been reported after spinal cord contusion, the effect of residual spinal cord displacement on reperfusion of blood flow or recovery of neurologic function remains unclear. METHODS: Eighteen beagles were anesthetized, and the spinal cord at T13 was loaded dorsally under precision loading conditions until evoked potential amplitudes were reduced by 50%. At this function endpoint, spinal cord displacement was maintained for 90 minutes. Somatosensory-evoked potentials were measured at regular intervals until 3 hours after decompression. Regional spinal cord blood flow was measured with a fluorescent microsphere technique at regular time points during and after spinal cord decompression. RESULTS: Within 5 minutes after dynamic cord compression was discontinued, evoked potential signals were absent in all dogs. Evoked potential recovery was observed after decompression in 7 of 18 dogs. Regional spinal cord blood flow at baseline, 21.8 +/- 1.9 mL/100 g. min (mean +/- SE), decreased to 3.9 +/- 0.9 mL/100 g. min after dynamic compression was discontinued. Although spinal cord-piston interface pressure dissipated by 87% of maximum interface pressure during sustained compression, mean blood flow recovered to only 34% of baseline flow. In the 7 dogs that recovered evoked potential function, blood flow increased to 11.3 +/- 2.7 mL/100g. min immediately before decompression (P < or = 0.05). In the 11 dogs that did not recover evoked potential function after decompression, regional blood flow did not improve during sustained compression. CONCLUSIONS: Recovery of evoked potential function after decompression corresponded with a greater return of blood flow during sustained displacement and greater reperfusion of blood flow associated with decompression. PMID- 10806498 TI - Topography and severity of axonal injury in human spinal cord trauma using amyloid precursor protein as a marker of axonal injury. AB - STUDY DESIGN: Axonal injury was examined in 18 human cases of acute spinal cord compression using amyloid precursor protein as a marker of AI. OBJECTIVES: To topographically map and semiquantitate axonal injury in spinal cord compression of sufficient severity to produce para- or quadriplegia. SUMMARY OF BACKGROUND DATA: Amyloid precursor protein is carried along the axon by fast axoplasmic transport and has been extensively used as a marker of traumatic axonal injury. METHODS: The study group comprised 18 cases of spinal cord compression (17 due to fracture dislocation of the vertebral column and 1 iatrogenic compression from Harrington rods) and two normal control. All the cords were examined according to a standard protocol, and at least 10 segmental levels were immunostained using a monoclonal antibody to amyloid precursor protein and immunopositive AI was semiquantitated using a grading system to provide the axonal injury severity score (AISS). The focal injury at the site of cord compression (haemorrhage, haemorrhagic necrosis, ischaemic necrosis) was also semiquantitated to provide the focal injury area score (FIAS). AI occurring around the site of focal compression (focal axonal injury severity score or FAISS) was distinguished from AI distant to the focal injury (nonfocal axonal injury severity score or NFAISS). RESULTS: All 18 cases showed widespread amyloid precursor protein immunoreactive axonal injury and the AISS ranged from 28 to 60%. In all cases, the FAISS was greater than the NFAISS and there was a statistically significant relationship between the AISS and the FIAS. CONCLUSION: Acute spinal cord compression of sufficient severity to produce permanent paralysis causes widespread axonal damage that is maximal at the site of compression but also present throughout the length of the cord in segments far distant from the site of the focal injury. PMID- 10806499 TI - Management of type II dens fractures: a case-control study. AB - STUDY DESIGN: A case-control study of patients with isolated type II dens fractures treated with halo vest immobilization. OBJECTIVES: To evaluate age as a risk factor for failure of halo immobilization in patients with type II dens fractures. SUMMARY OF BACKGROUND DATA: The literature reports an average fusion rate of approximately 70% in patients with type II dens fractures treated by halo vest immobilization. Although many investigators have examined patient age as a risk factor for nonfusion using halo immobilization, all studies have been supported only by Class III data. These studies, consequently, carry little or no statistical significance. Therefore, a case-control study based on Class II data was designed to evaluate age as a risk factor for failure of halo vest immobilization in patients with type II dens fractures. METHODS: Thirty-three patients with isolated type II dens fractures treated with halo vest immobilization at the University of Iowa Hospitals and clinics between 1983 and 1997 were included. Type II fractures were defined with plain radiography as per the Anderson-D'Alonzo classification. Cases were defined as nonfusions after halo immobilization, whereas control subjects represented successful bony unions attained with halo immobilization. RESULTS: When the case and control groups were compared, there was no significant difference between the groups in the presence of concomitant medical conditions, sex, the amount of fracture displacement, the direction of fracture displacement, the length of hospital stay, or length of follow-up. Age more than 50 years was found to be a highly significant risk factor for failure of halo immobilization (P = 0.002; Fisher's exact test, two tailed). The odds ratio of these data indicate that the risk of failure of halo immobilization is 21 times higher in patients aged 50 years or more. CONCLUSIONS: Surgical intervention should be considered in those patients aged 50 years or more who have a type II dens fracture, if it can be accomplished with acceptable risk of morbidity and death. PMID- 10806500 TI - The cervical facet capsule and its role in whiplash injury: a biomechanical investigation. AB - STUDY DESIGN: Cervical facet capsular strains were determined during bending and at failure in the human cadaver. OBJECTIVE: To determine the effect of an axial pretorque on facet capsular strains and estimate the risk for subcatastrophic capsular injury during normal bending motions. SUMMARY OF BACKGROUND DATA: Epidemiologic and clinical studies have identified the facet capsule as a potential site of injury and prerotation as a risk factor for whiplash injury. Unfortunately, biomechanical data on the cervical facet capsule and its role in whiplash injury are not available. METHODS: Cervical spine motion segments were tested in a pure-moment test frame and the full-field strains determined throughout the facet capsule. Motion segments were tested with and without a pretorque in pure bending. The isolated facet was then elongated to failure. Maximum principal strains during bending were compared with failure strains, by paired t test. RESULTS: Statistically significant increases in principal capsular strains during flexion-extension loading were observed when a pretorque was applied. All measured strains during bending were significantly less than strains at catastrophic joint failure. The same was true for subcatastrophic ligament failure strains, except in the presence of a pretorque. CONCLUSIONS: Pretorque of the head and neck increases facet capsular strains, supporting its role in the whiplash mechanism. Although the facet capsule does not appear to be at risk for gross injury during normal bending motions, a small portion of the population may be at risk for subcatastrophic injury. PMID- 10806501 TI - Morphometric analysis of thoracic and lumbar vertebrae in idiopathic scoliosis. AB - STUDY DESIGN: Prospective study on the morphometry of 337 pedicles in 29 patients with idiopathic scoliosis. OBJECTIVES: To analyze by means of computed tomographic scans the vertebral morphometry in idiopathic scoliosis treated by pedicle screw instrumentation. SUMMARY OF BACKGROUND DATA: Although several studies exist on the vertebrae's morphometry in normal spines, little is known concerning the morphometry of scoliotic vertebrae. METHODS: The pedicles' morphometry between T5 and L4 was analyzed by computed tomographic scans in 29 surgically treated patients with idiopathic right thoracic scoliosis. Measurements included chord length, endosteal transverse pedicle width, transverse pedicle angle, and pedicle length. RESULTS: The endosteal transverse pedicle width was significantly smaller (P < 0.05) on the concavity in the apical region of the thoracic spine and measured between 2.5 and 4.2 mm in the middle thoracic spine (T5-T9) and between 4.2 and 5.9 mm in the lower thoracic spine (T10-T12). In the lumbar spine, the width varied between 4.8 and 9.5 mm without significant differences between the concave and convex sides (P > 0.05). The chord length was shortest at T5, measuring 37 mm and increased gradually to 50 mm at L3 with significantly larger dimensions in male patients and on the concavity of the apical region in the thoracic spine (P < 0.05). The pedicle length varied minimally, with a range of between 20 and 22 mm, and was relatively consistent throughout the thoracic and lumbar spine. The transverse pedicle angle varied between 6 degrees in the lower thoracic spine and 12 degrees in the upper thoracic and lower lumbar spine. CONCLUSION: The morphometry in scoliotic vertebrae is substantially different from that of vertebrae in normal spines, with an asymmetrical intravertebral deformity shown in scoliotic vertebrae. Pedicle screw instrumentation on the concavity in the apical region of thoracic curves appears critical because of the small endosteal pedicle width. PMID- 10806502 TI - Prognosis for people with back pain under a No-fault 24-hour-cover compensation scheme. AB - STUDY DESIGN: A retrospective descriptive casenote review of consecutive back pain claimants assessing claim outcome at 12 months from onset. OBJECTIVE: To assess prognosis for back pain claimants in a no-fault 24-hour-cover accident compensation system. BACKGROUND: New Zealand has a unique accident compensation system that may provide incentives for health professionals to classify people with backache as having a back injury and incentives for back pain claimants to continue claims longer than would be the case in other compensation systems. METHODS: One hundred consecutive back pain claimants were identified from a single office of New Zealand's sole accident compensation insurer (Accident Rehabilitation and Compensation Insurance Corporation; ACC), who were still receiving compensation payments 4 weeks after the initial date of the claim. The study end point was case closure in the subsequent 12 months. Case closure rate was analyzed in relation to several potential prognostic variables. RESULTS: Of the 100 cases identified in which the claimant was receiving compensation 4 weeks from the initial date of the claim, 43 cases were not closed by 6 months, and 30 cases were not closed at 12 months. The variable most strongly associated with case nonclosure was whether the claimant was receiving earnings-related compensation (equal to 80% of previous income), with 41% of this group still receiving compensation at 12 months versus 16% of the group not receiving earnings-related compensation (chi2 = 8.55, P = 0.003). These results compare unfavorably with those from previous published studies from The Netherlands and Jersey in the United Kingdom. CONCLUSION: New Zealand's unique accident compensation environment may discourage return to work for people with back pain. New Zealand legislators should assess the impact of the ACC scheme on people with back pain, particularly in light of the recent recommendations of the International Association for the Study of Pain Task Force on Back Pain in the Workplace, that compensation cover for workers with back pain be limited to 6 weeks. PMID- 10806503 TI - Association between recovery outcomes for work-related low back disorders and personal, family, and work factors. AB - STUDY DESIGN: Incident cases of work-related low back disorders were identified in an automotive metal stamping plant. Individuals were asked to recall health and work habits at the time of diagnosis of the work-related disorder and to report their current low back pain and physical functioning. OBJECTIVES: To evaluate five measures of health outcomes for work-related low back disorders in an industrial population and to determine potentially modifiable correlates of recovery. SUMMARY OF BACKGROUND DATA: The factors that influence recovery in actively working adults and how to best measure recovery outcome in this population are not well known. METHODS: Eighty-eight hourly employees of a metal stamping plant who experienced a work-related low back disorder were interviewed. Health status, health habits, and family and work relations were assessed with a structured interview to determine their association with various recovery outcomes (low back pain, low back pain disability, physical functioning, general physical health, and lost workdays). Interview information was supplemented with data from the plant's Occupational Safety and Health Administration Form 200 log. RESULTS: The clinical measures of recovery from the work-related low back disorders examined had similar overall predictive ability. However, in multivariate analyses, different potentially modifiable prognostic variables emerged as significant among them. Poorer self-rated health status and high personal stress were correlated with low back pain disability. Higher levels of cigarette smoking were correlated with higher levels of low back pain disability, lower physical functioning, and more severe low back pain at follow-up. CONCLUSION: The choice of measure of recovery from work-related low back disorders should be made in the context of the rehabilitation intervention goal. Interventions designed to modify and promote healthful personal behavior should be given more emphasis in rehabilitation efforts for work-related low back disorders. PMID- 10806504 TI - Injuries to the lateral femoral cutaneous nerve during spine surgery. AB - STUDY DESIGN: A prospective study to locate patients with injured lateral femoral cutaneous nerve after elective spine surgery. OBJECTIVES: To assess the prevalence of injury of the lateral femoral cutaneous nerve and to identify the cause of injury according to the position of the patients at surgery and the surgical approach. SUMMARY OF BACKGROUND DATA: Injuries to the lateral femoral cutaneous nerve, also known as meralgia paresthetica, may cause pain and therefore result in restriction of activity. Compression of the nerve by disc hernia, retroperitoneal tumors, and external pressure around the anterior superior iliac spine are among the more common causes. METHODS: One hundred five patients admitted for elective spine procedures were grouped according to position on the operating table and surgical approach. All patients were examined before and after surgery for signs of injury to the lateral femoral cutaneous nerve, and those found injured were followed up for 1 year after surgery. RESULTS: Injury to the lateral femoral cutaneous nerve was found in 21 (20%) patients. In 6 of them, all of whom underwent surgery on the Hall-Relton frame, the injury was bilateral. In 7 patients the injury was not associated with discomfort. In addition to injury by external pressure at the anterior superior iliac spine from the Hall-Relton frame, the nerve was also injured at the retroperitoneum by hematoma or traction and at the anterior iliac crest when bone was harvested. In 89% of the patients, the nerve completely recovered within 3 months of surgery. Two patients still had pain 1 year after surgery and hypoesthesia of the anterolateral thigh. CONCLUSION: Injuries to the lateral femoral cutaneous nerve during spine surgery are frequent, and patients should be informed of the possible risk. It usually has a benign course, but some preventive steps should be taken: keep posterior to the anterior superior iliac spine and minimize retraction when harvesting a bone graft, pad the posts of the Hall-Relton frame over the anterior superior iliac crest, and avoid traction on the psoas muscle during the retroperitoneal dissection. PMID- 10806505 TI - Efficacy and validity of radiofrequency neurotomy for chronic lumbar zygapophysial joint pain. AB - STUDY DESIGN: A prospective audit. OBJECTIVE: To establish the efficacy of lumbar medial branch neurotomy under optimum conditions. SUMMARY OF BACKGROUND DATA: Previous reports of the efficacy of lumbar medial branch neurotomy have been confounded by poor patient selection, inaccurate surgical technique, and inadequate assessment of outcome. METHODS: Fifteen patients with chronic low back pain whose pain was relieved by controlled, diagnostic medial branch blocks of the lumbar zygapophysial joints, underwent lumbar medial branch neurotomy. Before surgery, all were evaluated by visual analog scale and a variety of validated measures of pain, disability, and treatment satisfaction. Electromyography of the multifidus muscle was performed before and after surgery to ensure accuracy of the neurotomy. All outcome measures were repeated at 6 weeks, and 3, 6, and 12 months after surgery. RESULTS: Some 60% of the patients obtained at least 90% relief of pain at 12 months, and 87% obtained at least 60% relief. Relief was associated with denervation of the multifidus in those segments in which the medial branches had been coagulated. Prelesion electrical stimulation of the medial branch nerve with measurement of impedance was not associated with outcome. CONCLUSIONS: Lumbar medial branch neurotomy is an effective means of reducing pain in patients carefully selected on the basis of controlled diagnostic blocks. Adequate coagulation of the target nerves can be achieved by carefully placing the electrode in correct position as judged radiologically. Electrical stimulation before lesioning is superfluous in assuring correct placement of the electrode. PMID- 10806506 TI - Cervical hemilaminectomy reconstruction: technical note. AB - STUDY DESIGN: A technique for posterior cervical hemilaminectomy reconstruction is described. OBJECTIVE: To describe a technique for reconstruction of cervical hemilaminae in an adult that resulted in osseous fusion and to suggest modifications for use in children. SUMMARY OF BACKGROUND DATA: Although uncommon, cervical hemilaminectomy can result in spine deformity. Re-establishment of the osseous and ligamentous structures should minimize this problem. METHOD: After excision of an intraspinal neurenteric cyst, removed cervical hemilaminae at C2 and C3 were reconstructed with metal plates, and posterior ligaments were reapproximated with suture. RESULTS: Radiologic evidence of osseous fusion was present at 1 year after surgery. CONCLUSIONS: The advantages of this technique are restoration of normal anatomy, technical ease, and simplicity. A modification of this technique in children by using absorbable rather than metal plates should allow for normal spine growth. PMID- 10806507 TI - Osteoid osteoma of the spine treated with percutaneous computed tomography-guided thermocoagulation. AB - STUDY DESIGN: Two cases are reported in which an osteoid osteoma of the lumbar spine was treated with CT-guided thermocoagulation. OBJECTIVES: To review an alternative and minimally invasive treatment for spinal osteoid osteomas. SUMMARY OF BACKGROUND DATA: Surgical resection of a spinal osteoid osteoma can, depending on the location, be a formidable undertaking. Bone scintigraphy can be helpful in intraoperative identification. More recently, resection through a computed tomography-guided drill hole was found to minimize exposure. Using a thermocoagulation probe, as has been used in osteoid osteoma of the extremities, may be technically easier and cause less morbidity. METHOD: With the patient under general anesthesia, a bone biopsy cannula was introduced into the center of the osteoid osteoma. Material was subjected to histologic examination. A thermocoagulation probe was then inserted and heated to 90 C for 4 minutes. The two patients were kept overnight for observation. RESULTS: Both patients had complete pain relief and no evidence of recurrence after 2 years' follow-up. There were no complications. Scoliosis resolved in one patient and persisted in the other. CONCLUSION: Percutaneous computed tomography-guided thermocoagulation is a minimally invasive and technically straightforward method to achieve ablation of a spinal osteoid osteoma. No complications were encountered in these two patients. Future research should focus on the safety of thermocoagulation, especially cephalad to the level of the conus medullaris. PMID- 10806508 TI - Enterobacter agglomerans spondylodiscitis: a possible, unrecognized complication of tetracycline therapy. AB - STUDY DESIGN: This case report describes infection in a lumbar disc in a healthy young man with an organism of low pathogenicity. The patient was taking a prolonged course of antibiotics at the time the infection occurred. OBJECTIVE: To describe this unique case of infective spondylodiscitis. SUMMARY OF BACKGROUND DATA: To the authors' knowledge, spinal infection with Enterobacter agglomeranshas never been reported. This organism is a transient gut colonizer, and may have established itself secondary to the patient's prolonged ingestion of tetracycline for acne. METHODS: This 22-year-old farmer had spontaneous lumbar back pain. Radiologic investigations showed an abnormality in the L4-L5 disc region, and together with other investigations, were suggestive of infection. The diagnosis was confirmed by surgical aspiration. RESULTS: Antibiotic therapy was administered, and the patient made a complete recovery. Follow-up radiographs showed a complete loss of the L4-L5 disc space with only minimal bone destruction. CONCLUSION: A unique cause of infective lumbar discitis is presented. Several features of this case are unusual. The magnetic resonance findings were not readily diagnostic. The cultured organism is usually nonpathogenic. The infection may have been secondary to prolonged tetracycline therapy. PMID- 10806509 TI - Treatment of cervical cord compression, caused by hereditary multiple exostosis, with laminoplasty: a case report. AB - STUDY DESIGN: Case report. OBJECTIVES: Successful excision of the exostosis within the spinal canal. SUMMARY OF BACKGROUND DATA: Myelopathy caused by exostosis within the spinal canal developed in a 13-year-old boy with hereditary multiple exostosis. METHODS: Spinous process-splitting laminoplasty with an ultrasonic knife was performed to remove the mass and minimize the possibility of postlaminectomy kyphosis. RESULTS: The spinal canal exostosis with cervical cord compression was excised successfully with laminoplasty. After surgery there has been no recurrence of tumor, and the stability of the cervical spine has been preserved. CONCLUSION: This is the first report of laminoplasty as a useful surgical approach for intraspinal exostosis to prevent postoperative cervical instability. PMID- 10806510 TI - Osteochondroma of the C5 lamina with cord compression: case report and review of the literature. AB - STUDY DESIGN: Case report of a solitary osteochondroma of the cervical spine causing myelopathy in a 66-year-old woman. OBJECTIVES: To review the relevant literature and describe a highly unusual clinical manifestation of solitary osteochondroma. SUMMARY OF BACKGROUND DATA: Osteochondromas are common benign bony lesions that seldom occur in the axial skeleton. These lesions are more commonly reported with neural compression in cases of hereditary multiple exostoses (Bessel-Hagel syndrome, diaphyseal aclasis). METHODS: Chart review, review of relevant radiographic examinations and histopathologic specimens, clinical follow-up with examination, and literature review. RESULTS: Manifestation with new neurologic deficit in a 66-year-old patient was singular. CONCLUSIONS: Osteochondromas are unusual in the axial skeleton, and are rarely signaled by neural compression. Occurrence is generally in young adults in the second and third decades. Initial manifestation with a new neurologic deficit in a 66-year-old patient was highly unusual. PMID- 10806511 TI - Dumbbell hydatid cyst of the spine: case report and review of the literature. AB - STUDY DESIGN: A case report of a hydatid cyst in the retropleural space at T7-T8 with an extension into the spinal canal through an intervertebral foramen complicated by paraparesis due to thoracic cord compression. OBJECTIVE: To confirm that hydatid cyst should be considered in the differential diagnosis for any mass discovered in the human body. SUMMARY OF BACKGROUND DATA: Appearance of a dumbbell-shaped mass in the thoracic spine is highly suggestive of neurofibroma. Hydatid cyst, although very rare, may have the same appearance. The spine is involved in 50% of hydatid disease of the bone. A hydatid cyst of the bone and muscle is always primary. Neurologic recovery is possible because it is a slow-growing lesion. METHOD: Clinical symptoms, differential diagnosis, and treatment are reviewed. The patient was treated successfully by total surgical excision of the lesion through a posterior retropleural approach. No medical treatment was prescribed. RESULTS: Surgical excision is effective in the treatment of hydatid disease of the spine, particularly if there is no extensive bony destruction and it is possible to achieve neurologic recovery after decompression of the thoracic spine. CONCLUSION: Extra spinal hydatidosis may lead to spinal cord compression by extension through the intervertebral foramen. The possibility of cure is high when there is no bony involvement. PMID- 10806512 TI - Symptomatic cervical synovial cyst associated with an os odontoideum diagnosed by magnetic resonance imaging: case report and review of the literature. AB - STUDY DESIGN: The first case of a synovial cyst of the upper cervical spine associated with os odontoideum diagnosed by magnetic resonance imaging is presented. OBJECTIVES: To evaluate distinct magnetic resonance imaging findings of a cervical synovial cyst located anterior to the cord and discuss its association with os odontoideum. These findings may guide further investigations. BACKGROUND: Synovial cysts of the spine are uncommon findings. They occur mostly dorsolaterally and with greatest frequency in the lumbar spine and are rarely symptomatic. No association with os odontoideum has been reported before. METHODS: A single subject with symptoms of deterioration in his hand functions was examined by a 2. 0-T magnetic resonance imager. RESULTS: Magnetic resonance imaging of the cervical spine showed an os odontoideum connected to the body of C2 by a synovial joint that had a cystic extension posteriorly compressing the cord. CONCLUSION: Synovial cysts should be considered in the differential diagnosis of an extradural mass of the upper cervical spine. Magnetic resonance imaging should be the choice in the investigation of such cases. PMID- 10806513 TI - Hypertrophic dens resulting in cervical myelopathy: histologic features of the hypertrophic dens. AB - STUDY DESIGN: A case report of a 43-year-old woman who had hypertrophic dens in the developmentally narrow atlas ring that resulted in cervical myelopathy. OBJECTIVES: To present histologic findings of the hypertrophic dens, which was excised en block the transoral approach, and to discuss the pathogenesis of the hypertrophic change of the dens. SUMMARY OF BACKGROUND DATA: Few cases have been reported of cervical myelopathy associated with hypertrophic dens, and there have been no previous reports describing the histologic features of hypertrophic dens. METHODS: Clinical data were reviewed from the patient's chart, and histologic features of the hypertrophic dens were examined in the sagittally cut section. RESULTS: After posterior decompression surgery, cervical myelopathy in the patient subsided. Myelopathy reappeared 4 years after surgery. Imaging studies showed osteoarthritis of the atlantodental joint, hypertrophic dens, and degeneration of the cruciate ligament. In the second surgery, transoral removal of the dens with posterior occipitocervical fusion was performed. Histologic studies showed thickening of the cortical bone of the anterior and cranial parts of the dens. The apical portion, which was the insertion portion of the ala ligaments, showed degenerative changes of tide marks. CONCLUSION: Atlantoaxial instability and enthesopathy were probably the causes of the hypertrophic changes of the dens in this case. PMID- 10806514 TI - Spine update: potential applications of gene therapy to the treatment of spinal disorders. PMID- 10806515 TI - Unknown case PMID- 10806516 TI - Re: is occupational low back pain on the rise? PMID- 10806518 TI - Imagery PMID- 10806517 TI - Re: posterior occipitocervical reconstruction using cervical pedicle screws and plate-rod systems. PMID- 10806519 TI - Our association on the worldwide web. PMID- 10806520 TI - "Recommendations for uniform reporting of data following major trauma--the Utstein style" (as of July 17, 1999). An International Trauma Anaesthesia and Critical Care Society (ITACCS). AB - Basic and advanced care of trauma patients has always been an important aspect of prehospital and immediate in-hospital emergency medicine, involving a broad spectrum of disciplines, specialties and skills delivered through Emergency Medical Services Systems which, however, may differ significantly in structure, resources and operation. This complex background has, at least in part, hindered the development of a uniform pattern or set of criteria and definitions. This in turn has hitherto rendered data incompatible, with the consequence that such differing systems or protocols of care cannot be readily evaluated or compared with acceptable validity. Guided by previous consensus processes evolved by the ERC, the AHA and other International Organizations--represented in ILCOR--on 'Uniform reporting of data following out-of-hospital and in-hospital cardiac arrest--the Utstein style' an international working group of ITACCS has drafted a document, 'Recommendations for uniform reporting of data following major trauma- the Utstein style'. The reporting system is based on the following considerations: A structured reporting system based on an "Utstein style template" which would permit the compilation of data and statistics on major trauma care, facilitating and validating independent or comparative audit of performance and quality of care (and enable groups to challenge performance statistics which did not take account of all relevant information). The recommendations and template should encompass both out-of-hospital and in hospital trauma care. The recommendations and template should further permit intra- and inter-system evaluation to improve the quality of delivered care and identification of the relative benefits of different systems and innovative initiatives. The template should facilitate studies setting out to improve epidemiological understanding of trauma; for example such studies might focus on the factors that determine survival. The document is structured along the lines of the original Utstein Style Guidelines publication on 'prehospital cardiac arrest'. It includes a glossary of terms used in the prehospital and early hospital phase and definitions, time points and intervals. The document uses an almost identical scheme for illustrating the different process time clocks--one for the patient, one for the dispatch centre, one for the ambulance and, finally, one for the hospital. For clarity, data should be reported as core data (i.e. always obtained) and optional data (obtained under specific circumstances). In contrast to the graphic approach used for the Utstein template for pre- or in hospital cardiac arrest, respectively, the present template introduces, for the time being, at least, a number of terms and definitions and a semantic rather than a graphic report form. The document includes the following sections: The Section Introduction and background The Section on Trauma Data Structure Development: presents a general outline of the development of structured data using object-orientated modelling (which will be discussed in due course) and includes a set of explanatory illustrations. The Section on Terms and Definitions: outlines terms and definitions in trauma care, describing different types of trauma (blunt, penetrating, long bone, major/combined, multiple/polytrauma and predominant trauma). The Section on Factors relating to the circumstances of the injury describes the following items: cause of injury (e.g. type of injury (blunt or penetrating), burns, cold, crush, laceration, amputation, radiation, multiple, etc. Severity of Injury e.g. prehospital basic abbreviated injury score developed by the working group. The score contains anatomical and physiological disability data, with the anatomical scale ranging ordinally from 1. Head to 9. External; the physiological disability scale ranging ordinally from 0--unsurvivable. Mechanism of injury recording for transportation incidents etc. e.g. the type of impact, po PMID- 10806521 TI - Effects of a hydroxyethylstarch solution on plasma colloid osmotic pressure in acutely ill patients. AB - Hydroxyethyl starch (HES) solutions can represent an alternative to human albumin solutions in intensive care unit (ICU) patients, but their effects on the plasma colloid osmotic pressure (COP) have not been well defined. We studied the changes in COP in 61 critically ill patients; 39 patients received 500 ml HES 6% (Elo Haes Fresenius solution) and 22 received 400 ml of a human albumin solution (4% albumin Belgian Red Cross) over 60 to 90 min. COP was determined with an oncometer, using a semi-permeable membrane at 30 Kd, before the infusion, at the end of the infusion, and one hour and four hours after the end of the infusion. COP increased in the HES group from 20.7 +/- 3.1 to 22.5 +/- 3.1 mmHg (p < 0.05), and this increase in COP was sustained throughout the 4 hours of the study. COP did not increase in the human albumin group (from 19.5 +/- 2.4 to 19.9 +/- 2.0 mmHg, NS). Hence, this standard HES solution has greater effects on COP than natural colloids. In view of their lower costs, HES solutions can represent a valuable alternative to human albumin. However, it is necessary to consider the secondary effects of HES and the physiologic functions of albumin. PMID- 10806522 TI - Case reports in anesthesiology: their trend through 17 years. AB - The study objective was to delineate the trend of case reports (the simplest of the descriptive forms of study) in the anesthesiological literature by analyzing the frequency of publication, and citation, and especially of the place of citation of a sample of published case reports. It is our opinion that case report in anesthesia is particularly suitable for this specialty rather than for others and is often the first signal of a complication, an adverse event, an anesthetic problem in rare disease and alerts other anesthesiologists to the possibility of unexpected events. METHODS: We analyzed the case reports published on an Anesthesiological journal placed in the middle in term of Impact Factor, from January 1980 to December 1997. Citations of each case report were obtained using computer searches of the Science Citation Index (SCI). For each of these case reports we collected in a custom-designed data base the following data: year of publication, number of authors, number of citations per year, place of citation, type of article quoting the case report, number of self-citations, year of first citation. MAIN RESULTS: We identified 637 case reports and 1946 citations. The number of case reports increased through the years up to a peak in 1994-95 and the same trend was observed for citations and self-citations, the number of authors per case report was < or = 4 in 90.4%; 74.2% of total case reports cited were first cited within two years of publication, while 34.7% were never cited. The type of article quoting the case reports has been, in the majority of cases, an original article. CONCLUSIONS: The analysed case reports and the number of citations can give us information about the importance of a clinical situation at a particular time. PMID- 10806523 TI - Suxamethonium-induced rhabdomyolysis in a healthy middle-aged man. AB - A 43-year-old man developed rhabdomyolysis after uvulo- palatopharyngoplasty. After induction with thiopentone and suxamethonium the anesthesia was maintained with halothane. The patient responded to treatment and made an uneventful recovery. In earlier reports of rhabdomyolysis after general anaesthesia with halothane and suxamethonium almost all the patients had malignant hyperthermia (MH) or muscular dystrophy. About 50% of malignant hyperthermia patients carry a mutation in the RYR1 gene. Our patient did not have mutations in the four MH associated genes tested, but the total amount of different mutations is by now about twenty. Therefore, despite these negative tests rhabdomyolysis may be a sign of subclinical malignant hyperthermia which cannot be ruled out by our investigations. This rare case of rhabdomyolysis in a healthy man suggests careful monitoring of the patient when-ever suxamethonium is used. PMID- 10806524 TI - Miss-'n-mix and mimics. Left-over ampules are stored in the local pharmacy stock. PMID- 10806525 TI - Possible postsynaptic action of aminoglycosides in the frog rectus abdominis. AB - The present study was undertaken to investigate the postsynaptic effects of aminoglycosides on contractions evoked by acetylcholine (ACh), KCl, electrical field stimulation (EFS) and Na(+)- and Ca(2+)-free Ringer solution with 0.2 mM Na2 EDTA (NaFCaFR) in the isolated frog rectus abdominis. Neomycin inhibited contraction elicited by ACh, NaFCaFR, and EFS at the higher frequencies (8 and 10 Hz) but not those elicited by KCl and EFS at the lower frequencies (2, 3 and 5 Hz). D-tubocurarine inhibited ACh-induced contractions in a concentration dependent manner. In addition, drug reduced EFS-evoked contractions to a limited extent. Lower concentrations (10(-5), 5 x 10(-5), 10(-4), 2 x 10(-4) and 3 x 10( 4) M) but not higher concentrations (4 x 10(-4) and 5 x 10(-4) M) of methoxyverapamil exhibited a concentration-dependent inhibitory action on NaFCaFR induced contractions. Similar inhibitions of the same type of contraction were displayed by aminoglycosides (neomycin, streptomycin, netilmycin, gentamycin and amikacin). These results suggest that in addition to their antagonistic action on nicotinic receptors in the frog rectus abdominis, aminoglycosides may exert stabilizing effects on some functional components contributing to contractions at the membrane. PMID- 10806526 TI - Purification, identification and phosphorylation of annexin I from rat liver mitochondria. AB - Annexin was purified from rat liver mitochondria to an apparent homogeneity with a molecular weight of 35 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified mitochondrial annexin (AXmito) was identified as annexin I by an immunoblot analysis using anti-annexin I antibody. The inhibitory effect of AXmito I on porcine pancreatic phospholipase A2 activity was as potent as that of bovine lung annexin I. The presence of annexin I in mitochondria was confirmed by an electron-microscopic study. AXmito I was shown to be phosphorylated by intrinsic protein tyrosine kinases on its tyrosine residues. This annexin was also phosphorylated by protein kinase C. PMID- 10806527 TI - Increased osteocyte apoptosis during the development of femoral head osteonecrosis in spontaneously hypertensive rats. AB - We investigated the presence of osteocyte apoptosis in the necrotic trabeculae of the femoral head of spontaneously hypertensive rat (SHR) using the in situ nick end labeling (TUNEL) method and transmission electron microscopy. The occurrence of osteonecrosis and ossification disturbance was significantly higher in SHR compared with Wistar Kyoto (WKY) rats, and Wistar (WT) rats used as control animals (P < 0.01). A high population of TUNEL positive osteocytes was detected mainly in 10- and 15-week-old SHRs. Sectioned examination of the femoral head of SHRs and WKY rats by electron microscopy revealed apoptotic cell appearances such as aggregation of chromatin particles and lipid formation. In contrast, a positive reaction was significantly lower in osteocytes in the femoral heads of WT rats (P < 0.01). Our results indicate that apoptosis forms an important component of the global pathologic process affecting the femoral head of SHR, which leads to osteonecrosis in this region. PMID- 10806528 TI - Women's anxiety in old age and long-term care provision for the elderly. AB - The purpose of this study was to verify the differences in women's anxiety in old age, the expected long-term care provision, and the expected final location for terminal care for the women themselves and for their parents. In addition, we examined factors that related to their anxiety and needs. The subjects were 1,000 women of the Seikatsu Club customer cooperative association in Chiba; 539 responded to our survey. The subjects were more anxious for their parents than for themselves. They more strongly expected long-term care for their parents to be provided by their family than they expected the same for themselves. Although no differences were observed in the expected location for terminal care, most subjects expected their home to be the terminal location. Analysis by the multiple logistic regression model indicated that the following factors were significantly related to the anxiety in old age: age odds ratio [OR = 1.81], employment [OR = 2.25] for women, and planning to live with parents [OR = 2.42], housing conditions [OR = 0.56] for parents. The following factors were significantly related to the expected long-term care provision: age [OR = 2.22] for women, and age [OR = 2.15], living with parents [OR = 3.58], and employment [OR = 2.33] for parents. Age [OR = 2.14] for women, and planning to live with parents [OR = 2.09] for parents were significantly related to the expected final location of terminal care. This survey showed that women expected long-term care for their parents to be provided by their family, while many expected public long term care services for themselves. This is the biggest difference in women's outlook on long-term care for their parents and for themselves. Multivariate analysis suggested that women aged 40 years or over, who will need long-term care in the future, tended to expect public home care services for themselves. It is virtually certain that the demand for public home care services will increase in the future. PMID- 10806529 TI - Measurement of fatigue in knee flexor and extensor muscles. AB - In order to examine fatigue of the knee flexor and extensor muscles and to investigate the characteristics of muscular fatigue in different sports, a Cybex machine was used to measure muscle fatigue and recovery during isokinetic knee flexion and extension. Eighteen baseball players, 12 soccer players and 13 marathon runners were studied. Each subject was tested in the sitting position and made to perform 50 consecutive right knee bends and stretches at maximum strength. This was done 3 times with an interval of 10 min between each series. The peak torque to body weight ratio and the fatigue rate were determined in each case. In all subjects, the peak torque to body weight ratio was higher for extensors than flexors. Over the 3 trials, the fatigue rate of extensors showed little change, while that of flexors had a tendency to increase. In each subject, knee extensors showed a high fatigue rate but a quick recovery, while knee flexors showed a low fatigue rate but a slow recovery. As the marathon runners had the smallest fatigue rates for both flexors and extensors, we concluded that marathon runners had more stamina than baseball players and soccer players. PMID- 10806530 TI - Creation and use of a composite polyurethane-expanded polytetrafluoroethylene graft for hemodialysis access. AB - The Thoratec (Vectra) polyurethane vascular access graft (TPVA) is among the most recent additions to the list of materials used to construct prosthetic grafts for vascular access during hemodialysis. We give the TPVA very high marks, and recognize the utility of such a graft for use in hemodialysis. However, the strong elasticity of this graft can lead to unexpected complications after suturing. We devised a new surgical method using a TPVA-ePTFE (expanded polytetrafluoroethylene) composite graft, substituting the anastomosis section of the TPVA with a portion of ePTFE graft material, and have been able to overcome most of the TPVA's potential problems. We herein describe the technique. PMID- 10806531 TI - President's message. Partners in care--the magic continues. PMID- 10806532 TI - Extending nursing care into the world of technology. PMID- 10806533 TI - Affective competencies. PMID- 10806534 TI - Syms Memorial Operating Pavilion. PMID- 10806535 TI - New testing method may detect heart disease earlier. PMID- 10806536 TI - Minimally invasive treatment of twin-to-twin transfusion syndrome. AB - Selective laser photocoagulation of communicating vessels (SLPCV) is an application of minimally invasive endoscopic fetal surgery (i.e., surgical fetoscopy) performed for severe cases of twin-to-twin transfusion syndrome (TTTS), a complication of monochorionic twin pregnancies. Advances in ultrasound and endoscopy have aided the identification and treatment of this potentially lethal or fatal condition. Surgical fetoscopy, an innovation in the field of surgery and maternal-fetal medicine, benefits patients by reducing the morbidity and mortality associated with TTTS, with minimal risks to the mother. PMID- 10806537 TI - Laparoscopic splenectomy for splenic sequestration crisis. AB - Medical and surgical advances have improved the treatment of splenic sequestration crisis in pediatric patients with sickle cell disease (SCD). Rapid enlargement of the spleen can result from sickled blood cels being trapped in the spleen, which can be life threatening. The laparoscopic splenectomy procedure using the lateral approach has been adapted successfully for the pediatric patient since 1993. Children with SCD who have a history of a splenic sequestration crisis usually are scheduled for an elective splenectomy procedure after the first documented crisis to reduce the risk of death. Some key benefits of this new surgical approach include a shortened hospital stay, decreased postoperative pain, and faster recovery and return to normal activities for the child with SCD. PMID- 10806538 TI - Osteochondral autotransfer--newer treatment for chondral defects. AB - Osteoarthritis is believed to be accelerated by full thickness defects of articular cartilage. This article describes emerging therapies, including mosaicplasty autologous (i.e., the patient's own) graft and autologous chondrocyte implantation techniques. Patients with chondral defects caused by a variety of possible pathologies experience positive implications, including return to normal function and delaying the onset of osteoarthritis. PMID- 10806539 TI - Clinical practice guidelines--an organizational effort. AB - In response to the national trend toward developing clinical practice guidelines, AORN set out to create practice guidelines specific to perioperative nursing. In this effort, a designated group of individuals began defining and reviewing guidelines from other organizations and determining parameters for developing AORN's own patient-focused, comprehensive, research-based guidelines. Clinical practice guidelines now have been published in AORN's Standards, Recommended Practices, and Guidelines, and a template has been established to help other AORN members create and implement guidelines for clinical conditions, common procedures, or patient populations specific to perioperative nursing practice. PMID- 10806540 TI - Effect of compensation and patient scheduling on OR labor costs. AB - To determine whether to accept a contract to provide additional surgical cases, OR managers must determine the incremental costs of caring for the new patients. The expected profitability of the contract can be computed by subtracting the incremental costs from the revenue. For surgical procedures, the incremental costs of OR labor significantly depend on how employees are paid (e.g., part-time versus full-time). If a surgical suite employs full-time staff members, incremental labor costs also are affected by how the day and time of patients' cases are selected (e.g., whether new cases are scheduled weeks in advance by the surgeon and the patient, or are performed on short notice based on the discretion of the surgical suite). This article explains how to estimate the incremental costs of staffing an OR for a case and discusses the use of internet-based online exchanges to match demand for OR time for additional cases to available unused OR capacity in variety of surgical suites. PMID- 10806541 TI - Older adults and HIV. AB - As the number of older Americans grows, perioperative nurses increasingly will be called on to provide services to older adults infected with HIV. To provide quality patient care, perioperative nurses must understand the facts about HIV and aging individuals. Finally, perioperative nurses must realize the best defense against exposure to bloodborne pathogens (e.g., HIV) is compliance with standard precautions while caring for all patients, from newborns to older adults. PMID- 10806542 TI - The multistate licensure compact. PMID- 10806543 TI - The educational activity approval process. PMID- 10806544 TI - Internet resources for researching twin-to-twin transfusion syndrome. PMID- 10806545 TI - Learn more using focus groups. PMID- 10806546 TI - [Chemical reactions catalyzed by DNA polymerases]. AB - Chemical reactions catalyzed by various DNA polymerases are discussed, including DNA chain extension, the 3'-->5'-exonuclease proofreading activity, and some other pathways of replicative repair. The contribution of DNA polymerases to the fidelity of the template-dependent synthesis is analyzed by the examples of some most typical DNA polymerases. PMID- 10806547 TI - [The relationship between the biological functions of sphingolipids and their chemical structure]. AB - The interrelation between the bioeffector functions of sphingolipids and their chemical structure is reviewed. The effects of modifications of sphingoid functional groups on the bioregulatory properties of sphingolipids in cell proliferation, differentiation, and apoptosis are discussed. PMID- 10806548 TI - [Photoactivated derivatives of peptide and protein ligands in the studies of neuroreceptors]. AB - The application of photoactivatable derivatives for studies of different types of neuroreceptors belonging to two superfamilies, G-protein dependent receptors and ligand-gated ionic channels, is discussed. Studies of the structure of voltage gated and ion-gated channels with the use of specific photoactivatable derivatives of neurotoxins are also described. Possibilities and prospects for the application of photoactivatable ligands in studies of the spatial structure of neuroreceptors are reviewed. PMID- 10806549 TI - [Analytical biotechnology of recombinant peptides and proteins. I. Analysis of the purity, composition and structure of human, porcine and bovine insulins]. AB - A method for analysis of the type, purity, and possible structural modifications of insulins of bovine, porcine, and human origin was proposed. It is based on a combination of narrow-bore reversed-phase HPLC and mass spectrometry. The hydrolysis of insulins with highly specific Glu-protease V8 from Staphylococcus aureus followed by peptide mapping of the hydrolysis products and mass spectrometry of the isolated fragments helps rapidly and reliably localize and identify substitutions of amino acid residues in insulin structure by using insulin samples of less than 1 nmol. PMID- 10806550 TI - [Hormone-like activity of a synthetic decapeptide with the adrenocorticotropin like sequence of human immunoglobulin G1]. AB - The antiproliferative and immunosuppressive in vitro effects of immunocortin, a synthetic adrenocorticotropin-like (ACTH-like) decapeptide H-Val-Lys-Lys-Pro-Gly Ser-Ser-Val-Lys-Val-OH, whose sequence corresponds to segment 11-20 of the variable part of the human IgG1 heavy chain, were studied. At concentrations of 10(-11)-10(-7) M, immunocortin was found to inhibit the growth of the human MT-4 T-lymphoblastoid cell line, to suppress the blast transformation of thymocytes, and to decrease the spontaneous mobility of peritoneal macrophages and their bactericidal action toward the virulent strain Salmonella typhimurium 415. By using a 125I-labeled "addressing" fragment of ACTH ?[125I]ACTH-(13-24)?, we showed that MT-4 cells express specific receptors for ACTH (Kd 97 pM). Immunocortin and human ACTH (but not the heavy chain of IgG1) competitively inhibited the binding of [125I]ACTH-(13-24) to these receptors with Ki1 of 0.38 and Ki2 of 0.34 nM, respectively. Specific receptors for ACTH (Kd 5.8 nM) on mouse thymocytes were detected and characterized. The unlabeled immunocortin was shown to complete with labeled ACTH-(13-24) for binding to these receptors (Ki = 1.8 nM) and this binding of immunocortin to receptors on thymocytes activates adenylate cyclase from these cells and increases the intracellular concentration of cAMP. PMID- 10806551 TI - [Synthesis and fluorescent properties of 5-(1-pyrenylethynyl)-2'- deoxyuridine containing oligodeoxynucleotides]. AB - Novel reagents for the fluorescent labeling of oligo- and polynucleotides have been prepared: 5-(1-pyrenylethynyl)-2'-deoxyuridine 3'-phosphoramidite and a solid support carrying this nucleoside. Oligonucleotides containing one or several modified units have been synthesized, and the fluorescence of these probes has been shown to change upon hybridization with the complementary sequence. PMID- 10806552 TI - [The degradation of glycoproteins with lithium borohydride. Isolation and analysis of O-glycopeptides with reduced C-terminal amino acid residue]. AB - By the example of fetuin and a blood-group-specific mucin from porcine stomach, we showed that, under conditions of reductive degradation of glycoproteins with LiBH4-LiOH in 70% aqueous tert-butyl alcohol, the reduction and cleavage of amide bonds occur much faster than the simultaneous beta-elimination of carbohydrate chains O-linked with Ser and Thr residues of the peptide chain. The major degradation products containing the O-linked glycans are the O-glycosylated derivatives of 2-aminopropane-1,3-diol and 2-aminobutane-1,3-diol (the products of reduction of glycosylated Ser and Thr) and the glycopeptides containing 2-4 amino acid residues with reduced C-terminal amino acid. Seventeen homogeneous O glycopeptides were isolated from the fetuin degradation products by ion-exchange and reversed-phase HPLC. Their structures were determined by MALDI-TOF mass spectrometry and by analyses for amino acids, amino alcohols, and carbohydrates. The application of the reaction for characterization of O-glycans and localization of O-glycosylation sites in O- and N,O-glycoproteins is discussed. PMID- 10806553 TI - [The isolation, preliminary study of structure and physiological activity of water-soluble polysaccharides from squeezed berries of Snowball tree Viburnum opulus]. AB - Water-soluble polysaccharide fractions VO1-VO4 were isolated from the squeezed berries of snowball tree (Viburnum opulus) by successive extraction with water at various temperatures and pH and with aqueous solutions of ammonium oxalate. These fractions were purified by ion-exchange chromatography on DEAE cellulose, and the homogeneity of the purified polysaccharides was determined by gel filtration on Sephacryl S-500. Acidic polysaccharides close to pectins in their sugar composition were found in all the extracts (fractions VO1-1, VO2-1, VO3-2, and VO4-2). Residues of galacturonic acid, galactose, arabinose, and (to a lesser extent) rhamnose are their main constituents. Neutral polysaccharides composed mainly of galactose and mannose residues were additionally found in fractions extracted with acidified water (pH 4.0) and with aqueous ammonium oxalate solutions. Partial acidic hydrolysis and digestion with pectinase of acidic polysaccharides indicated that their carbohydrate backbone consists of alpha-1,4 linked residues of D-galacturonic acid. NMR spectra of acidic polysaccharides (fractions VO3-2 and VO3-3) confirmed this and demonstrated that their side oligosaccharide chains are composed of beta-1,4-linked galactopyranose residues and of terminal and 2,5- and 3,5-substituted residues of alpha-arabinofuranose at a Gal: Ara ratio of 3:1. Some polysaccharides from V. opulus were found to possess an immunostimulating activity: they enhance phagocytosis, in particular, the phagocytic index and the secretion of lysosomal enzymes with peritoneal macrophages. Calcium ions were found to be necessary for the appearance of the stimulating effect of acidic polysaccharides from V. opulus. PMID- 10806554 TI - [The inhibition of complement-dependent hemolysis of liposomes containing cerebroside sulfate]. AB - Cerebroside sulfate (CGS) was found to be capable of inhibiting complement dependent hemolysis. The activity dependence of CGS-containing liposomes on their composition was studied. Mixtures of CGS with phosphatidylethanolamine, phosphatidylserine, sphingomyelin from cattle brain, cerebroside from cattle spinal cord (CG), and egg yolk phosphatidylcholine (ePC) were investigated. In the case of binary CGS/ePC mixtures, the antihemolytic activity varied nonlinearly with an increase in the mass part of CGS: it sharply increased with an increase in the CGS part from 0.3 to 0.5 and decreased by 20-30% of the maximum value with an increase in the CGS part from 0.9 to 1. On the basis of these experiments, the optimum distance between the charged groups of CGS was estimated to be 0.92-1.6 nm. In the ternary compositions of 4:3:3 CGS/ePC/polar lipid, only CG increased the activity of liposomes as compared to that of liposomes from the 4:6 CGS/ePC. The preliminary incubation of CGS-containing liposomes with complement decreased hemolysis more effectively than incubation with other components of the hemolytic system. This suggests that the interaction of CGS-containing liposomes with the complement proteins is responsible for their antihemolytic activity. PMID- 10806555 TI - [Oligo(2'-O-tetrahydropyranylribonucleotides): hybridization properties and nuclease resistance]. AB - Oligo(2'-tetrahydropyranylribonucleotides) and their analogues containing a 3'-3' internucleotide bond at the 3'-terminus are nuclease-resistant and possess rather high affinity toward RNA, the main target in the antisense approach. PMID- 10806556 TI - Blood stasis, thrombosis and fibrinolysis. AB - Blood stasis is a very common occurrence associated with many of our daily activities. Prophylaxis against development of thrombosis in high-risk conditions is well established. In this issue of Hematology/Oncology Clinics of North America, international experts present new and provocative information on the basic mechanisms, diagnosis, and management of blood stasis, thrombosis, and fibrinolysis. Reviews of pertinent topics by three physicians in training also are included. I hope that the information provided will expand our understanding of this interesting subject. PMID- 10806557 TI - Imaging in blood stasis. The role of imaging techniques in defining the causes, presence, and effects of blood stasis. AB - There have been considerable developments in the ability to image blood vessels and blood flow using ultrasound, CT, and MR imaging. The effects of vascular pathology in causing changes in the blood leading to thrombus formation now can be seen clearly because of these developments. In particular, new ultrasound MR imaging techniques allow more precise assessment of vessel walls and flow than has ever been possible before using conventional techniques, such as angiography. MR imaging has a unique potential for noninvasively demonstrating the natural history of developing vascular disease and the effects of this on blood flow and progression to thrombosis. PMID- 10806558 TI - The fibrinolytic enzyme system. Basic principles and links to venous and arterial thrombosis. AB - This article briefly describes some important aspects of the fibrinolytic system, its regulation, and possible disturbances of this system in connection with deep vein thrombosis and myocardial infarction. PMID- 10806559 TI - A comprehensive review of vitamin K and vitamin K antagonists. AB - For more than 60 years, vitamin K-dependent proteins have been known to play an important role in regulating blood coagulation. During recent years it has become clear, however, that vitamin K is also involved in other physiologic processes, including bone metabolism and vascular biology. Because the vitamin K requirement of bone and vessel wall is higher than that of the liver (where the clotting factors are produced) recommended daily allowance (RDA) values for K vitamins must be redefined. According to the new definition, a substantial part of the population is mildly deficient in vitamin K, and at later ages this deficiency may contribute to increased bone fracture risk, arterial calcification, and cardiovascular disease. PMID- 10806560 TI - Hypercoagulable states. AB - The hypercoagulable state has been defined as the potential to develop thrombosis in association with hereditary and noninherited genetic mutations and acquired disorders. It is a condition that places an individual at risk for, but does not in itself inevitably lead to, thrombosis. The focus of this article is understanding mechanisms in the hypercoagulable state that enhance and maintain the production of thrombin in circulating blood while preventing its progression to thrombosis. These mechanisms include reactions that produce thrombin from prothrombin, feedback loop mechanisms that affect the rate of thrombin production from prothrombin and the inactivation of thrombin in blood. The fibrinolytic system is involved in clot lysis but not in thrombin production and inactivation. PMID- 10806561 TI - Screening for risk factors for thrombosis using a new generation of assays developed to evaluate the functionality of the protein C anticoagulant pathway. AB - These results, obtained in a small series of patients, suggest that both the ProC Global assay and the PCP Test would be suitable, using well-defined cut-off levels, to identify all the carriers of the Factor V Leiden mutation and all the patients with a protein C deficiency or with combined defects of the protein C pathway. For both assays, however, the sensitivity for protein S deficiency was below 60%. These results in selected patients are congruent with those previously reported in the literature about the ProC Global assay, the PCP Test, and other assays evaluating the functionality of the protein C anticoagulant pathway. All demonstrated a weak sensitivity to protein S deficiency, suggesting that protein S plays only a minor role as an APC cofactor in such global assays. A major discrepancy between the two evaluated assays was obtained in the group of patients without abnormality of the protein C pathway. Actually, using the ProC Global assay, more than 40% of the patients had a decreased PCAT-NR while presenting with none of the three tested abnormalities, whereas none of the studied patients had a ratio below 1.80, and only 5 of 143 (3.5%) had a ratio below 2.00 when using the PCP Test. The observation that around 40% of the control patients had a decreased PCAT-NR could suggest the influence of currently unknown defects of the protein C/protein S pathway on the ProC Global assay. It could also be hypothesized that the higher factor VIII levels already reported in patients with a history of thrombosis than in controls had a significant role in the low responsiveness, but this parameter was not tested in the authors' series. In that connection, it is also well established that elevated factor VIII levels both shorten the APTT and reduce the anticoagulant effect of heparin when evaluated using APTT. Actually, some of the samples investigated in this study were obtained during the acute phase of thrombosis. It is not possible to draw out the hypothesis of an association of biologically undetectable minor changes in various factors involved in the protein C anticoagulant pathway; all the individual factors would remain within their normal ranges. Finally, because 40% of the patients without abnormalities of the protein C pathway had a decreased PCAT-NR, the question arises whether the ProC Global assay might in itself be a biologic marker of thrombophilia, independent of its sensitivity for abnormalities of the protein C anticoagulant pathway. In that connection, the correlation between the result of the ProC Global assay and the risk for thromboembolism was recently evaluated by two different groups. In both cases, the preliminary results suggested that a decreased response to the ProC Global assay might be an independent risk factor for venous thrombosis. The two global assays could therefore have distinctly different applications. If the global assays are used in the hemostasis laboratory to screen for abnormalities of the protein C pathway, and thus to rationalize the use of specific assays, the PCP Test should be chosen, because of its high specificity. Because only 3.5% of the control patients had a ratio below 2.00 (and none had a ratio below 1.80), the PCP Test could be accurately used as a first-step assay in the laboratory screening for these abnormalities of the protein C anticoagulant pathway. Using such a flow chart, the specific assays for APC resistance or the identification of the factor V Leiden mutation and protein C would be performed only in case of a ratio below a cut-off defined using receiver operating characteristic (ROC) analysis in unselected patients. Because of the weak sensitivity of this assay to both constitutional and acquired protein S deficiencies (below 15% using 1.80 as the cut-off level, or 60% using 2.00), the measurement of this parameter had to be performed in all cases. If, on the other hand, the assay is used to screen for risk factors for thrombosis, the ProC Global assay could b PMID- 10806562 TI - Risk factors for venous thromboembolism following prolonged air travel. Coach class thrombosis. AB - Venous thromboembolism (VTE) in legs and lungs is a potentially life-threatening condition. The incidence of VTE associated with air travel is still unknown, but it may have increased. Most travelers who develop symptoms do so within 24 hours after their flight takes off. Predisposing risk factors may be divided into patient-related and cabin-related factors, both of which are described. It is emphasized that better information and better inflight precautions can minimize these risk factors. PMID- 10806563 TI - Mechanisms of thrombosis in spinal cord injury. AB - Many studies failed to identify a hypercoagulable imbalance in the blood factors or decreased anticoagulant activity. On the other hand, fibrinolysis, a process unrelated to hypercoagulability but closely related to endothelial cell integrity, is predictably altered and contributes to the persistence of venous occlusion by thrombosis. There is considerable evidence that interruption of neurologic impulses and the ensuing paralysis cause metabolic changes in blood vessels and that blood vessel changes are accountable for venous thrombosis. Altered venous competence with complete spinal cord injury manifests by a decrease in venous distensibility and capacity and an increase in venous flow resistance. Vascular adaptations to inactivity and muscle atrophy, rather than the effect of a nonworking leg-muscle pump and sympathetic denervation, seem to lead to the thrombosis; indicating that thrombosis resulting from venous incompetence cannot be reversed by anticoagulation alone. PMID- 10806564 TI - Thrombosis risk in the trauma patient. Prevention and treatment. AB - Hypercoagulability is frequently seen in the trauma patients. Debate continues over the best method of prophylaxis, diagnosis, and treatment for the trauma patient. From experience with orthopedic and general surgery patients, much has been learned about prophylaxis and diagnosis, and as treatment protocols have been taken from internal medicine literature. Universal guidelines relating specifically to the trauma patient have not, however, been established. Overall, most of the literature suggests using LMWH for the prophylaxis of trauma patients. When LMWH is contraindicated, SCD should be used, with AVFP as a second choice. Surveillance screening for DVT remains controversial, but surveillance before transfer to extended care facilities has proven beneficial. Finally, when DVT is diagnosed, treatment should be initiated as soon as possible and should be continued until the DVT has resolved. Long-term anticoagulation therapy or use of caval filters may be necessary to prevent the morbidity of PE or thrombophlebitic syndrome. PMID- 10806565 TI - Thromboembolic complications associated with reproductive endocrinologic procedures. AB - Thromboembolism as a complication of hormonal ovarian stimulation in the context of artificial reproductive techniques is rare and seems to occur when OHS is present. Although accompanied by high serum estrogen concentrations, hCG seems to play a central role in the development of OHS, which has been observed in women with 17,18-desmolase deficiency who have low estrogen levels after induction of ovulation with hGC. Although there is some evidence that hormonal ovarian stimulation with HMG, leading to elevated estrogen levels, and ovulation induction with hCG in preparation for in vitro fertilization are associated with a state of hypercoagulability, the exact role of estrogens, hCG and the physicochemical changes (fluid shift into third spaces) involved in OHS remain to be elucidated. PMID- 10806566 TI - Clinical application of argatroban as an alternative anticoagulant for extracorporeal circulation. AB - The authors attempted experimental and clinical use of argatroban as an alternative anticoagulant in left heart bypass with the centrifugal pump, percutaneous cardiopulmonary support (PCPS), and extracorporeal membrane oxygenation (ECMO) to determine if it has complementary effects in preventing thrombus formation without aggravating bleeding tendency. Its reversible binding to thrombin and its short half-life contributed to reduce the risk of excessive blood loss without clot formation within the extracorporeal circulation circuit during thoracic aortic surgery using left heart bypass. PCPS and ECMO were safely performed at doses ranging from 0.5 to 10 micrograms/kg/min to maintain activated clotting time at approximately 200 seconds. Although experimental studies showed argatroban to be advantageous in preserving platelet and fibrinogen, further clinical investigations are necessary. PMID- 10806567 TI - Safety of recombinant and plasma-derived medicinals for the treatment of coagulopathies. AB - The introduction of advanced technologies (PCR testing, chromatography, and specific viral inactivation and removal techniques) has led to remarkable improvements in the quality and efficacy of biopharmaceutical products. The current safety strategies for both recombinant protein and PDP products depend on the extensive screening of the source material for infectious agents and the use of mild purification methods, specific mild viral-reduction techniques, GMP, QC, and QA. An appropriate system of pharmacologic vigilance is also an integral element for assuring product quality and safety in the marketplace. Such precautions make available high quality therapeutic recombinant proteins and PDP products. The risks in the clinical setting and the cost/benefit ratio must be considered in choosing a product for therapeutic use. The choice should be based on the analysis of data available for a specific product, because some variations in quality and safety can be observed in different brands. Overall, a much finer control of infectious risks has been achieved, and improvement will continue. With the new products, thrombotic episodes have become rare. Reducing immunogenic potential and improving yield to increase product supply could be the next challenges for producers of biopharmaceuticals. PMID- 10806568 TI - Indications for surgical treatment of iliofemoral vein thrombosis. AB - The goals of treatment of acute iliofemoral DVT should be prevention of fatal PE, reduction of pain and swelling of the involved leg, trying to stop the development of phlegmasia cerulea dolens and venous gangrene, prevention of disabling PTS by early removal of the blood clot, avoiding proximal venous obstruction, preserving normal, functioning valves in the leg veins, and preventing reflux. The authors recommend an aggressive approach with rapid removal of the occluding thrombus in the leg veins extending up into the iliac veins in active patients with a short history of symptomatic DVT, usually less than 7 days. This approach is not justified in chronically ill, bedridden, high risk, or aged patients, or those with serious intercurrent disease or limited life expectancy. In these patients, such interventions can only be indicated for limb salvage in phlegmasia cerulea dolens when conservative treatment does not prevent the development of an acute compartment syndrome with venous gangrene. The preferred means of accomplishing early and quick removal of the thrombus is CDITT. Most of the authors' positive experience with thrombolysis is based on the use of urokinase. The Food and Drug Administration (FDA) has put this drug on temporary hold for almost 1 year. The alternative drugs (e.g., tissue plasminogen activator [tPA]) have not been tested for CDITT of DVT, and tPA is not FDA approved for this indication. When there are contraindications or failure of thrombolysis, TE with a temporary AVF is a valid alternative. PMID- 10806569 TI - Statin drugs and dietary isoprenoids as antithrombotic agents. AB - Statin drugs and various isoprenoids from plant origins inhibit mevalonic acids, cholesterol, and other isoprenoid products. Among these, reduction of farnesyl and geranylgeranyl prenylated proteins impedes signal transduction at the cellular level. The authors envision that limiting such prenylated proteins downregulates thrombin-stimulated events, including decreasing the expression and availability of protease-activated receptor-1 mitigating thrombin stimulation of cells, tissue factor preventing additional thrombin generation, and plasminogen activator inhibitor-1 allowing thrombosis. Additional processes may enhance nitric oxide production and induce other processes. Downregulation of thrombin stimulated events should promote hypothrombotic or quiescent conditions that reduce cardiovascular disease, thus contributing to longevity. PMID- 10806570 TI - [A clinical study of patients undergoing curative surgery for renal pelvic and ureteral cancers]. AB - We retrospectively studied 30 patients who underwent curative surgery for renal pelvic and/or ureteral cancer between August 1987 and August 1998. Their clinicopathological features were classified by the criteria of the Japanese Urological Association. The 1-, 3-, and 5-year cause-specific survival rates were, respectively, 100, 95.5, and 85.1%, while the disease-free rates were 100, 78.9, and 78.9% by the Kaplan-Meier method. Prognostic factors were evaluated by the log-rank test. The significant prognostic factors were pT3 and pV1 for cause specific survival (p = 0.0277, p = 0.0025), while pT2 (or higher), grade 3, and pV1 were significant for disease-free survival (p = 0.0271, p = 0.0327, and p = 0.0002). Nine patients who received adjuvant chemotherapy are alive, but 3 patients have relapsed. Chemotherapy did not have a significant effect on the cause-specific survival or disease-free survival. PMID- 10806571 TI - [Antireflux surgery in infants with vesicoureteral reflux]. AB - Febrile urinary tract infections (UTI) resulted in the admission of five infants (four boys, one girl) to Okinawa Prefectural Hospital in Miyako. The first febrile episode developed at an average age of 1.3 months. All patients underwent cystoscopy and X-ray studies to role out secondary causes for vesicoureteral reflux (VUR). All patients had high grade reflux and underwent antireflux surgery using a modification of the Amar-Paquin method or the Politano-Leadbetter method. They tolerated the procedures without any complications. No UTI recurred and VUR disappeared in one year following surgery. Early surgical intervention seems beneficial for selected infants with high grade VUR, but the surgical procedure and methods for infants, especially those with severe dilated ureter, are still controversial. PMID- 10806572 TI - [A clinical study on active vitamin D pulse therapy in hemodialysis patients with secondary hyperparathyroidism]. AB - The oral active vitamin D pulse therapy was performed for 20 chronic hemodialysis patients with secondary hyperparathyroidism (2 degrees HPT). Before pulse therapy, all patients showed high-turnover bone diseases, elevated serum alkaline phosphatase (Alp) and elevated serum parathyroid hormone (PTH). They had at least one parathyroid gland detected by ultrasonography. In one patient, serum PTH level did not decrease during three months. In five patients, hypercalcemia was observed so that the pulse therapy was with drawn. Another patient was resistant to the second pulse therapy for recurrence of 2 degrees HPT. Total parathyroidectomy and autotransplantation was performed in these seven patients. The size of parathyroid gland in patients undergoing surgical treatment was significantly larger than those estimated by ultrasonography in patients successfully treated with pulse therapy (p < 0.005). In all patients with surgical treatment, the conventional oral active vitamin D therapy was performed with caution to hypercalcemia preoperatively. The preoperative Alp levels decreased compared with those of at the beginning of the pulse therapy. Postoperatively, the total amount of intravenous administered calcium decreased, and serum calcium levels were easily controlled. PMID- 10806573 TI - [A case of angiosarcoma in retroperitoneal cavity]. AB - We report a case of angiosarcoma in the retroperitoneal cavity. A 66-year-old man was admitted to our hospital with the complaint of chest discomfort. Imaging studies including computed tomography scan and magnetic resonance imaging revealed a large solid retroperitoneal mass adjacent to the bladder. There was no evidence of metastasis. The tumor was surgically resected and histopathologically diagnosed as angiosarcoma from the positive staining for VIIIth factor, mitotic figures and abnormal endothelial cells. Six months after surgery, local recurrence adjacent to the bladder appeared. We resected the mass and started adjuvant therapy using Interleukin-2. However, he died of progressed disease one year after the first operation. PMID- 10806574 TI - [A case of retroperitoneal schwannoma: diagnostic usefulness of magnetic resonance imaging]. AB - A 36-year-old man visited our hospital with a chief complaint of difficult urination. He was admitted for further examination because ultrasonography showed a heterogeneous solid mass in the lower abdomen. Intravenous urography showed the right ureter and bladder to be pushed leftward without hydronephrosis. Computerized tomography showed a giant solid mass in the retrovesical space. Magnetic resonance imaging revealed a round, well-encapsulated tumor, showing low intensity on the T1-weighted image, heterogeneously high intensity on the T2 weighted image and irregular enhancement with Gd-DTPA. Histopathology of the extirpated tumor was benign schwannoma. PMID- 10806575 TI - [Cyclophosphamide-induced renal pelvic tumor--a case report]. AB - We report a case of transitional cell carcinoma in the left renal pelvis, which occurred in a 24-year-old man. He had been treated with cyclophosphamide (CPM) for a period of 27 months for retroperitoneal rhabdomyosarcoma diagnosed at the age of 10. At first 1.2 g CPM had been given twice intravenously for 3 months, followed by oral administration of 41 g CPM for 23 months. Drip infusion pyelography revealed a filling defect in the left renal pelvis. A left renal pelvic tumor was strongly suspected on computerized tomography and magnetic resonance imaging. Left nephroureterectomy was then performed. Histological diagnosis of the left renal pelvic tumor was transitional cell carcinoma, grade 2, pT1N0M0. No recurrence was defected 17 months later. This case seems to be the second case of cyclophosphamide-induced upper urothelial carcinoma reported in Japan. PMID- 10806576 TI - [Hydronephrosis as a complication of the appendiceal abscess: report of a case]. AB - We report a case of appendiceal abscess complicated with right hydronephrosis in a 67-year-old woman. She visited our department complaining of right flank pain. Intravenous pyelography (IVP) showed hydronephrosis and hydroureter on right and a narrow ureter at the pelvic brim. A pelvic computed tomographic scan revealed a low density area measuring 44 x 37 mm in size, anterior to the right ureter, which was thought to be a pelvic tumor. Further examination did not reveal the origin of the tumor. An exploratory laparotomy was performed. The tumor developed from the cecum and adhered to the right ureter. The appendix was not found. The tumor was resected with the cecum while the ureter was preserved. Histological investigations revealed an appendiceal abscess. The postoperative course was uneventful. IVP after the operation showed the hydronephrosis to have resolved. PMID- 10806577 TI - [Crossed fused kidney with ectopic ureter: a case report]. AB - A 5-year-old girl with a rare anomaly of a single system ectopic ureter draining into the mesonephric duct cyst and ipsilateral crossed fused kidney is presented. Both ureters were catheterized and three dimensional (3-D) images of the urinary tract were reconstructed with helical computed tomographic (CT) data. A mesonephric duct cyst, the right ureter draining into the cyst and an ectopic orifice were well depicted with these images, especially with inferior projection images. 3-D display of helical CT data with a retrograde contrast study is a promising method of obtaining anatomical detail of an ectopic ureter. PMID- 10806578 TI - [A case of epithelial membrane antigen-positive leiomyosarcoma of the urinary bladder]. AB - A 41-year-old woman was admitted with gross hematuria and pain on urination. Cystoscopy showed a huge and lobulated submucosal non-papillary bladder tumor. Pelvic computed tomography demonstrated a heterogeneous and enhanced lobulated mass, 8 cm in diameter, with extravesical invasion but there appeared to be no metastatic lesions. Transurethral biopsy revealed leiomyosarcoma pathologically. Total cystectomy and construction of an ileal conduit were performed. The tumor was histologically diagnosed as leiomyosarcoma. Immunohistochemical studies revealed the tumor to be positive for epithelial membrane antigen (EMA) and muscle actin but negative for desmin, and S-100. We reviewed 102 cases of vesical leiomyosarcoma reported in Japan. Among these 102 cases, there were no EMA positive cases. Immunohistochemical and electromicroscopic evaluation should be performed to evaluate this disease. PMID- 10806579 TI - [Sarcomatoid carcinoma of the urinary bladder: report of a case]. AB - The patient was a 78-year-old man with gross hematuria. A non-papillary and non pedunculated tumor was found on the posterior wall of the bladder by cystoscopic examination. Total cystectomy with construction of ileal conduit was performed. Histologically, the tumor was composed of carcinomatous and sarcomatous elements. The carcinomatous element was compatible with grade 3 transitional cell carcinoma. The sarcomatous element was composed of osteosarcomatous, chondrosarcomatous pattern and undifferentiated malignant spindle cell component. Immunohistochemical examination demonstrated the presence of keratin, cytokeratin and epithelial membrane antigen in both carcinomatous and sarcomatous elements. Therefore, we diagnosed this tumor as sarcomatoid carcinoma. The patient has remained well without any evidence of recurrence for 10 months after operation. PMID- 10806580 TI - [Primary AA type amyloidosis of the urinary bladder: a case report]. AB - Primary amyloidosis of the urinary bladder is a rare disease entity. A total of 61 cases have been reported in the Japanese literature, and most of them were AL type amyloidosis. We report here a case of primary AA type amyloidosis. A 52-year old man presented with a chief complaint of asymptomatic gross hematuria. Cystoscopy revealed yellowish elevated lesions, transurethral mucosal biopsies were performed, and the histopathological diagnosis indicated a primary AA type amyloidosis of the urinary bladder. Systemic amyloidosis was clinically eliminated. The yellowish lesions in the bladder through cystoscopy disappeared spontaneously one year later without any specific treatment, but periodical work up may be necessary to rule out recurrence of the disease or bladder tumor. PMID- 10806581 TI - [Malignant mesothelioma presenting as a perineal and pelvic mass--a trial of high dose intra-arterial methotrexate injection and irradiation]. AB - A 50-year-old male with the complaints of lumbago and voiding disturbance was diagnosed to have malignant mesothelioma. Serum CA-125 was found to be elevated. The tumor was stained positive immunohistochemically only for CA-125 and epithelial membrane antigen. Magnetic resonance imaging of the pelvic demonstrated a large mass extending from the right external obturator muscle to the perineum. He was treated by two courses of methotrexate given intra arterially (2,000 mg) followed by external beam irradiation at a total dose of 60 Gy. Disease progression was not apparent 15 months after treatment. PMID- 10806582 TI - [A case of female paraurethral cyst diagnosed as epithelial inclusion cyst]. AB - The patient was a 36-year-old woman with a chief complaint of pain of the urethral meatus. A cyst existed just below the urethral meatus. The paraurethral cyst was removed completely. Histologically, the cyst was lined by stratified squamous epithelium. According to the criteria proposed by Das, paraurethral cysts are classified into 4 groups: epithelial inclusion cysts, Mullarian cysts, Gartner duct cysts, Skene duct cysts. In this case, the cyst seemed to be an epithelial inclusion cyst. In the Japanese literature, paraurethral cysts have been incompletely classified from an etiological perspective. Differential diagnoses based on the histological findings of the cysts may be important to determine the etiology. PMID- 10806583 TI - [Primary testicular carcinoid tumor: a case report]. AB - This report describes a primary testicular carcinoid. A 41-year-old male was hospitalized with an asymptomatic right testicular mass. A high inguinal orchiectomy was done after the diagnosis of the testicular tumor. Pathologically, the tumor showed the typical appearance of a carcinoid tumor. A computed tomographic scan and other studies could not demonstrate any metastasis elsewhere. He has remained well and without any evidence of recurrence. PMID- 10806584 TI - Preimplantation genetic diagnosis--an integral part of assisted reproduction. PMID- 10806585 TI - Assessment of maximal fertilization rates with intracytoplasmic sperm injection. AB - PURPOSE: Maximal fertilization rates following ICSI were assessed using two essential steps: immobilization of sperm and aspiration of oocyte cytoplasm. METHODS: ICSI procedure was performed for couples (N = 42) in whom the male suffered severe infertility or failure of fertilization in previous IVF cycle using different oocyte cytoplasmic aspiration and sperm immobilization methods. Outcome in four patient groups was measured by oocyte damage, fertilization rate, and pregnancy rate. RESULTS: Maximal fertilization (90%) were achieved from the group which used immobilization of sperm by hard-touching the tail with a pipette and optimal aspiration of oocyte cytoplasm. CONCLUSIONS: The results suggest that if the immobilization of sperm and aspiration of oocyte cytoplasm are handled right during ICSI, this procedure can be expected to yield a 90% fertilization rate. PMID- 10806586 TI - A simplified ultrasound based infertility investigation protocol and its implications for patient management. AB - PURPOSE: To evaluate whether a simplified infertility investigation protocol, focusing on the use of hysterocontrast sonography (HyCoSy), one blood test, and a semen analysis, would be sufficient as an initial screening test to select couples for specific treatment. METHODS: The infertile couples underwent gynaecological examination, cervical sampling for cytology and Chlamydia trachomatis culture, B-mode transvaginal ultrasonography and basic hormonal analyses followed by a HyCoSy, and a semen analysis. A preliminary diagnosis was made for all patients. A management plan for treatment was suggested when possible; otherwise further examinations were recommended. The data were stored for later analysis and the routine investigation protocol was then adhered to and a final diagnosis and treatment were decided upon. RESULTS: Agreement between the diagnosis based on HyCoSy and our routine protocol was present in 74% of cases (N = 73). In 13% (N = 13) there was partial agreement. In 36% the HyCoSy based protocol was considered sufficient to suggest treatment. CONCLUSIONS: A simplified approach may lead to a significant reduction in both the time and cost of investigating an infertile couple. PMID- 10806587 TI - Is diagnostic testicular fine needle aspiration necessary in azoospermic men before sperm aspiration/extraction for intracytoplasmic sperm injection cycles? AB - PURPOSE: To determine whether diagnostic testicular fine needle aspiration (TEFNA) sampling needs to be performed in azoospermic men prior to obtaining testicular sperm cells for IVF-ICSI procedures. METHODS: Ten azoospermic patients underwent TEFNA in 1993-1996. During 1997, all patients underwent testicular sperm aspiration (TESA) and/or testicular sperm extraction (TESE) to retrieve spermatozoa for IVF-ICSI cycles. The results of the two procedures performed in two separate hospitals were compared. RESULTS: Diagnostic TEFNA revealed spermatozoa in five patients; identical results in four were found during IVF ICSI cycles. In three patients, only Sertoli cells were found on TEFNA, in two of them TESA/TESE showed identical results, and in one, two spermatozoa were detected by Cyto-SEM. In the remaining two patients, spermatids or spermatocytes were found on both procedures. CONCLUSIONS: There was a very good correlation between the diagnostic and therapeutic procedures. We suggest that in azoospermic patients, diagnostic TEFNA is valuable in order to avoid unnecessary controlled ovarian hyperstimulation in the female partner for IVF. In patients in whom spermatozoa are detected, cryopreservation may be performed for later IVF-ICSI cycles. PMID- 10806588 TI - Sex chromosomal analysis of spermatozoa from infertile men using fluorescence in situ hybridization. AB - PURPOSE: To confirm an association between male infertility and chromosome aberrations of spermatozoa, we demonstrated the frequency of numerical abnormalities of spermatozoa from infertile men with abnormal semen parameters compared with fertile controls. METHOD: Sperm cells from 10 infertile patients were investigated for disomy rates of sex chromosomes and chromosome 18 and diploidy by fluorescence in situ hybridization (FISH). All patients showed oligoasthenozoospermia with sperm counts 3-20 x 10(6)/ml and motile rates 0-40%. RESULTS: Regarding XY disomy, a significantly higher frequency was found in 8 of 10 patients as compared to normal fertile men. The disomy rates of chromosome 18, XX, YY, and diploidy rate were not increased. CONCLUSIONS: There is an association between male infertility and embryo with aneuploidy of sex chromosomes. Counseling about possible genetic risks should be provided to the infertile couples planning assisted reproduction treatment. PMID- 10806589 TI - High progesterone levels and ciliary dysfunction--a possible cause of ectopic pregnancy. AB - OBJECTIVES: To investigate the effects of different levels of hormones on the ciliary activity of human oviducts and, consequently, to assess their possible role in tubal implantation of the fertilized egg. DESIGN: Fallopian tube epithelial samples were incubated in media with the addition of Estradiol (E2), progesterone (P), human menopausal gonadotropin (hMG), LH, or pure FSH (Metrodin) in different concentrations. The ciliary beat frequency (CBF) was measured after 24 h of incubation. Then the media were exchanged to media without the addition of hormones and the CBF was measured again 24 h later by using the photoelectric technique. SETTING: University teaching hospital, IVF unit. RESULTS: Twenty-four hr after the addition of P to the culture medium in concentrations of 0.5 or 1 ng/ml a significant decline of the CBF down to 63% of the control level was observed (P < 0.001) and with P in concentration of 2 ng/ml or greater, 50-70% of the cilia were paralyzed. These effects of P were found to be reversible. Incubation with E2 induced a slight increase of 4% in the mean CBF (P = 0.002). Twenty-four hr incubation with Metrodin, Pergonal, or LH did not affect ciliary motility. CONCLUSIONS: Higher levels of progesterone cause ciliary dysfunction and subsequently may be a possible cause of ectopic pregnancy. PMID- 10806590 TI - A randomized prospective cross-over study of highly purified follicle-stimulating hormone and human menopausal gonadotrophin for ovarian hyperstimulation in women aged 37-41 years. AB - PURPOSE: In the present study, we investigated the benefits of highly purified FSH (H.P. FSH) in comparison with human menopausal gonadotrophin (hMG) in IVF patients aged 37-41 years. METHODS: Twenty patients experienced within a period of four months both preparations in subsequent cycles through a prospective randomized cross-over design. A standard hormonal treatment consisting of a flare up protocol using gonadotrophin-releasing hormone agonist (GnRHa) in combination with the same starting dose of H.P. FSH or hMG was used in all cycles. RESULTS: Cycles stimulated with H.P. FSH resulted in a significantly higher mean number (10.3 +/- 3) of ovocytes retrieved with a significantly shorter (13 +/- 2.3 days) duration of stimulation compared to cycles treated with hMG [mean number of oocytes 7.3 +/- 5 (P < 0.01) and mean duration of stimulation 14.7 +/- 3.9 days (P < 0.02)]. No detrimental effect of basal or exogenous LH on oocyte quality was observed in our study. CONCLUSIONS: In our experience, H.P. FSH preparations seemed to be more effective than hMG preparations for ovarian stimulation in this group of women aged 37-41 years. PMID- 10806591 TI - Effectiveness of low dose of gonadotropin releasing hormone agonist on hormonal flare-up. AB - PURPOSE: The hormonal response (flare-up) following administration of a standard dose (100 micrograms) or a low dose (25 micrograms) of gonadotropin releasing hormone agonist (GnRH-a) (Triptorelin) was compared in patients prior to an in vitro fertilization (IVF) cycle and during the early follicular phase of a short term IVF protocol. METHODS: The gonadotroph (FSH, LH) and steroid [estradiol (E2) and progesterone (P)] flare-up was studied on two consecutive cycles in 30 normo ovulatory women. Patients were randomized to receive either 25 or 100 micrograms of triptorelin for three days at the beginning of the first cycle. Then doses were switched according to a crossing over design in the second cycle. RESULTS: No significant difference in the magnitude of FSH and E2 release could be observed following administration of the two doses of agonist whereas maximal plasma LH level was significantly reduced after injection of 25 micrograms of triptorelin. CONCLUSIONS: As compared to a standard dose, using a low dose of GnRH-a induces an hormonal flare-up which seems adequate for an optimal follicular recruitment. PMID- 10806592 TI - Cytokine levels in a patient with severe ovarian hyperstimulation syndrome before and after the ultrafiltration and reinfusion of ascitic fluid. AB - PURPOSE: To report serum concentrations of several cytokines (interleukin-6, interleukin-8, tumor necrosis factor-alpha, and vascular endothelial growth factor) before and after the reinfusion of ultrafiltrated ascitic fluid. METHODS: A case report of a woman hospitalized for the treatment of severe OHSS at the Department of Obstetrics and Gynecology, Tottori University Hospital. The serum concentrations of cytokines were analyzed by ELISA. RESULTS: Cytokine concentrations declined in parallel with the improvement of clinical conditions and resolution of OHSS. CONCLUSION: Measurement of serum cytokine concentrations may be useful in evaluating the severity of OHSS. PMID- 10806593 TI - Primer system for single cell detection of double mutation for Tay-Sachs disease. AB - PURPOSE: Nearly 100% of infantile Tay-Sachs disease is produced by two mutations occurring in the alpha chain of the lysosomal enzyme beta-N-acetylhexosaminidase (HEXA) in the Ashkenazi Jewish population. Although others have described primer systems used to amplify both sites simultaneously, few discuss the allele dropout problems inherent in this test. Our goal was to construct a more robust test enabling stronger signal generation for single cell preimplantation genetic diagnosis and to investigate the occurrence of allele dropout. METHODS: New nested primers were designed to optimize detection of both major Tay-Sachs mutations. Four hundred fifty-seven single cells, including normal cells and those carrying mutations of either the 4bp insertion exon 11 or splice-site intron 12 defects, were used to screen a new primer system. RESULTS: Based on PCR amplified product analysis, total efficiency of amplification was 85.3%, (390/457). The allele dropout rate for the 4bp insertion mutation in exon 11 and splice-site mutation in intron 12 was 4.8% and 5.8%, respectively. CONCLUSIONS: Multiple mutation detection and analysis within the Tay-Sachs disease gene (HEXA) is possible using single cells for clinical preimplantation genetic diagnosis. Alternative PCR primers and conditions offer various methods for developing systems compatible to specific program requirements. PMID- 10806594 TI - Understanding differences in the perception of anonymous parties: a comparison between gamete donors and their recipients. PMID- 10806595 TI - Review: drug therapy in Chinese traditional medicine. AB - This article addresses the differences between the basic concepts of drug therapy in Chinese traditional medicine (CTM) and modern Western medicine. In ancient times, throughout the world, humans used plants as food and medicine. In Europe, chemistry developed rapidly after the influence of Paracelsus. Active principles were isolated from plants, and drugs were prepared in the salt form to use as medication. In China, several drugs isolated from plants have been prepared in the salt form in recent years, but herbal medication, developed in the ancient tradition, continued to be widely used in Chinese populations. Some theories such as the yin-yang theory and the five-element theory were commonly employed to explain the rationale for the use of CTM. In this review, hypertension and diabetes mellitus are used as examples to illustrate the applications of CTM. The notions of the "whole" and the use of "relative" rather than absolute are important concepts, which distinguish CTM from modern medicine. PMID- 10806596 TI - Toxicity of complementary therapies: an eastern perspective. AB - Chinese traditional medicine is used extensively in Chinese populations, and other Asian countries employ similar therapies. Herbal treatments have a major role in these systems, and although most have a well-established safety record, occasional adverse effects are seen. Problems arise when toxic herbs are used in excessive doses, with improper preparation, or when they are substituted erroneously. There may also be adulteration with Western drugs or heavy metals, and interactions between herbs and Western drugs may also occur. It is always prudent to obtain a complete history of the use of herbal medications during any clinical assessment, particularly in Asian patients. PMID- 10806597 TI - Antioxidant effects of Chinese traditional medicine: focus on trilinolein isolated from the Chinese herb sanchi (Panax pseudoginseng). AB - It is thought that oxygen-derived free radicals (OFR) cause lipid peroxidation, which contributes to the process of atherosclerosis, and they are also involved in the myocardial damage seen with ischemia and reperfusion. Antioxidants could potentially ameliorate such harmful effects. Many natural plant products have been shown to have antioxidant effects. Trilinolein, a triacylglycerol purified from Panax pseudoginseng, which is commonly used in Chinese traditional medicine, has been found to have pharmacological effects, including antioxidant activity that may explain the benefits in treating circulatory disorders perceived from the use of the herb over the centuries. PMID- 10806598 TI - Alternative therapies and medical science: designing clinical trials of alternative/complementary medicines--is evidence-based traditional Chinese medicine attainable? AB - Evidence-based traditional Chinese medicine is attainable. With good planning and a positive attitude, the remedies used in traditional Chinese medicine (TCM) and Chinese proprietary medicines can be studied at a standard acceptable to modern science. The identification of an active principal should not delay the search for effective remedies from the TCM pharmacopoeia. Herbal mixtures can be validly tested to establish their efficacy. Problems with potential batch-to-batch variation can be circumvented by appropriate randomization. Subsequent independent screening and randomization to treatment and placebo arms can allow for the individualization of treatments by TCM practitioners. However, clearly defined treatments are required and should be recorded in a manner that enables other suitably trained researchers to reproduce them reliably (e.g., using prescriptions in Chinese). Quality control of TCM is a prerequisite of credible clinical trials. Correct natural ingredients must be used without adulteration or erroneous substitution. Evidence of safety in man is essential, and in lieu of data from formal toxicity studies, clear, convincing, and impartial evidence of safety is needed based on their long-term use in mainstream TCM practice backed up by publications in the Chinese medical/scientific literature. PMID- 10806599 TI - Rank power of metrics used to assess QTc interval prolongation by clinical trial simulation. AB - Monte Carlo simulation was used to assess the type I error rate and rank order of power for six different metrics using linear mixed-effect models, including two variables recommended by the European Agency for the Evaluation of Medicinal Products (EMEA) in the analysis of QTc interval data. The metrics analyzed were maximal change in QTc interval from baseline, maximal QTc interval, area under the QTc interval-time curve (AUC), average QTc interval, maximal QTc interval with baseline QTc interval as covariate, and AUC with baseline QTc interval as covariate. Two dosing regimens were studied: multiple-dose oral and multiple-dose continuous intravenous infusion. Both regimens were designed to produce similar maximal plasma concentrations, albeit with the infusion regimen maintaining maximal plasma concentrations for a longer period of time. The ability of the metrics to detect a drug effect was examined, assuming drug effect followed either an Emax or linear model. All statistics had a type I error rate near the nominal value. Regardless of pharmacokinetic or pharmacodynamic model, AUC with baseline QTc interval as a covariate had greater power than any other metric examined. The simulations also suggest that mean QTc interval data not be used. PMID- 10806600 TI - Pharmacokinetics, pharmacodynamics, and safety of microencapsulated octreotide acetate in healthy subjects. AB - The pharmacokinetics, pharmacodynamics, and safety of the marketed formulation of microencapsulated octreotide acetate were evaluated in an open-label study in 22 healthy cholecystectomized subjects. Each subject received a single 30 mg dose of microencapsulated octreotide acetate intramuscularly (i.m.). Concentrations of octreotide, growth hormone (GH), insulin-like growth factor binding protein 3 (IGFBP-3), and insulin-like growth factor 1 (IGF-1) as well as clinical safety were evaluated over a period of 112 days (16 weeks). After the injection, mean serum octreotide concentration initially increased rapidly, reached the maximum (Cmax, day 1 = 0.96 +/- 0.25 ng/ml) approximately 1.5 hours after dosing, and declined thereafter until 24 hours postdose (Cmin, 24 h = 0.088 +/- 0.093 ng/ml). The octreotide concentration then increased and started a sustained release from day 7 onward. Plateau concentrations were maintained through day 70 and gradually declined to below the lower limit of quantification (LLOQ) by day 112. The plateau height (Cplateau (2-112d, 60%)) was 1.68 +/- 0.88 ng/ml, and the duration (delta plateau, 60%) was 30.2 +/- 15.7 days. The integrated concentration-time curve, AUC0-112d, was 2819 +/- 782 (ng.h/ml), and the apparent half-life (t1/2) was 169 hours. To assess the variability, the drug concentrations were determined hourly for 8 hours on day 28, and the mean octreotide concentration, Cavg, day 28' was 1.55 +/- 1.26 ng/ml. The suppression of IGF-1 was statistically significant compared to the baseline (p < 0.05) through day 63; however, there were no appreciable changes in GH and IGFBP-3 concentrations after a single injection of microencapsulated octreotide acetate. Simulation of a 28-day dose schedule suggested that steady-state octreotide concentrations would be reached by the third injection with steady-state concentrations about twofold greater than the first injection. There were no serious adverse events or clinically meaningful changes in vital signs, ECGs, or laboratory evaluations observed in this study, indicating that the 30 mg i.m. dose of microencapsulated octreotide acetate was well tolerated in this population. PMID- 10806601 TI - Pharmacokinetics of single-dose reboxetine in volunteers with renal insufficiency. AB - Reboxetine is a new selective norepinephrine reuptake inhibitor (selective NRI) for the short- and long-term treatment of depression that is effective and well tolerated at a dose of 8 to 10 mg/day. This study assessed the pharmacokinetics of reboxetine in volunteers with renal impairment. A single 4 mg dose of reboxetine was administered to a total of 18 volunteers with mild (n = 6), moderate (n = 6), or severe (n = 6) renal impairment (creatinine clearance: 56 64, 26-51, and 9-19 ml/min, respectively), and reboxetine concentrations were measured in plasma by HPLC. Mean AUC infinity increased by 43% (mild vs. severe; p < 0.01) as renal function declined, while renal clearance and total urinary excretion of unchanged reboxetine decreased by 67% and 62%, respectively (mild vs. severe; p < 0.01 for both parameters). tmax and t1/2 were not significantly different between groups. In comparison with historical data from young healthy volunteers, AUC infinity and t1/2 are at least doubled in volunteers with renal impairment, while CLr is halved. This pharmacokinetic study has shown that increasing renal dysfunction leads to increasing systemic exposure to reboxetine, particularly in severe renal insufficiency, although reboxetine was well tolerated by all volunteers. Thus, a reduction of the starting dose of reboxetine to 2 mg twice daily would be prudent in patients with renal dysfunction. PMID- 10806602 TI - Pharmacokinetics and pharmacodynamics of sibrafiban, an orally administered GP IIb/IIIa antagonist, following coadministration of aspirin and heparin. AB - Sibrafiban is a double prodrug that is converted to the inactive single prodrug and to the active GP IIb/IIIa antagonist after oral administration. This clinical investigation evaluated whether coadministration of oral aspirin or intravenous heparin would alter the pharmacokinetics or pharmacodynamics of oral sibrafiban. Twenty-four adult subjects received two of the following four combinations: sibrafiban alone, sibrafiban with ASA, sibrafiban with heparin, and sibrafiban with ASA and heparin, separated by a 2-week washout period. Concentration profiles of active drug in citrate and EDTA plasma were unchanged with coadministration of ASA or heparin. No pharmacodynamic interaction was seen with coadministration of heparin. Inhibition of platelet aggregation increased 4% to 55%, and Ivy bleeding time increased 58% to 87% with coadministration of sibrafiban and ASA. The combined pharmacodynamic effect of sibrafiban and ASA may indicate a potentially greater therapeutic effect but an increased risk of bleeding when these drugs are used in combination. PMID- 10806603 TI - Antiplatelet effects of MK-852, a platelet fibrinogen receptor antagonist, in healthy volunteers. AB - MK-852, a cyclic heptapeptide, is a potent platelet fibrinogen receptor antagonist. When administered to normal healthy male subjects by 1- and 4-hour constant rate intravenous infusions, it provides a generally well-tolerated and reversible means of inhibition of platelet function. At infusion rates of 1 microgram/kg/min for 1 hour and 0.44 microgram/kg/min for 4 hours, respectively, MK-852 extended baseline bleeding time by greater than 2.2-fold and 2.6-fold, inhibited ADP-induced platelet aggregation by 76% and 69%, and inhibited collagen induced platelet aggregation by 65% and 67%, respectively. The pharmacokinetics of MK-852 include an elimination half-life of approximately 2 hours, total clearance of about 150 ml/min, and volume of distribution of about 18 liters. Examination of the relationship between MK-852 whole-blood concentration in vitro and inhibition of platelet aggregation showed an EC50 of about 55 ng/ml and a Hill coefficient of 1.55. The infusions were generally well tolerated, with no study drug-related changes in blood counts or biochemical profiles. PMID- 10806604 TI - Pharmacokinetics and pharmacodynamics of tenecteplase: results from a phase II study in patients with acute myocardial infarction. AB - Tenecteplase is a site-specific engineered tissue plasminogen activator (t-PA) variant that can be administered as a single intravenous bolus injection because of its slower plasma clearance. The objective of this study was to investigate the dose-ranging pharmacokinetics and pharmacodynamics of intravenous bolus tenecteplase compared with intravenous alteplase recombinant t-PA in patients with acute myocardial infarction. A total of 103 patients received intravenous bolus doses of 30, 40, or 50 mg tenecteplase, and 56 patients received 100 mg rt PA as the accelerated 90-minute infusion regimen in this randomized, open-label study. Tenecteplase and r-tPA plasma concentrations were measured for 6 hours. Tenecteplase exhibited biphasic elimination from the plasma with a mean initial half-life of 22 minutes and a mean terminal half-life of 115 minutes. The mean plasma clearance was 105 mL/min and did not depend on tenecteplase dose over the dose range studied. In comparison, rt-PA has a fourfold faster plasma clearance. Pharmacokinetic-pharmacodynamic evaluation showed that a dose of approximately 0.5 mg/kg results in a plasma AUC value that provides a TIMI 3 flow at 90 minutes that is comparable to that reported with accelerated r-tPA. In conclusion, tenecteplase has a fourfold slower plasma clearance compared with rt-PA, allowing dosing as an i.v. bolus injection. Weight-adjusted dosing of tenecteplase may optimize the therapeutic regimen of tenecteplase. PMID- 10806605 TI - A study comparing the clinical pharmacokinetics, pharmacodynamics, and tolerability of triamcinolone acetonide HFA-134a metered-dose inhaler and budesonide dry-powder inhaler following inhalation administration. AB - The impending phaseout of chlorofluorocarbons as propellants in pressurized metered-dose inhalers used in the treatment of asthma has resulted in the development of alternative devices to deliver drug to the pulmonary airways. These alternative devices include metered-dose inhalers using environmentally friendly hydroflurocarbon propellants and breath-actuated dry-powder inhalers. The purpose of this study was to compare the single- and multiple-dose pharmacokinetics, pharmacodynamics, and tolerability of a newly developed hydroflurocarbon formulation of triamcinolone acetonide (Azmacort HFA 225 mcg Inhalation Aerosol) to that of the dry-powder formulation of budesonide (Pulmicort Turbuhaler 200 mcg). This three-way crossover study used 18 normal healthy subjects each receiving a 675 mcg dose of triamcinolone acetonide, 600 mcg dose of budesonide, or placebo twice a day for 5 days. Serial plasma samples were collected after the first and last dose of test medication for pharmacokinetic analysis. Pharmacodynamics were assessed by changes in hypothalamic-pituitary-adrenal axis function as measured by 8 a.m. serum cortisol, 24-hour overnight serum cortisol AUC(0-24), and 24-hour urinary-free cortisol after the last evening dose of test drug. Tolerability was assessed through physical examinations, vital signs, 12-lead ECG, routine clinical labs, and adverse events recording. Both compounds were systemically absorbed. However, no significant drug accumulation was noted with chronic dosing. Chronic dosing did result in a statistically significant 20% reduction in basal 24-hour serum cortisol AUC(0-24) for both compounds. There were no clinically significant abnormalities in physical examination, vital signs, 12-lead ECG, or routine clinical labs noted during the study. Overall, the study drugs were well tolerated, with adverse events characterized as mild to moderate in severity. PMID- 10806606 TI - Coadministration of tacrolimus and mycophenolate mofetil in stable kidney transplant patients: pharmacokinetics and tolerability. AB - The tolerance and pharmacokinetics (PK) of tacrolimus (T) by the addition of mycophenolate mofetil (MMF) in stable kidney transplant patients (6/group) on long-term tacrolimus-based therapy were investigated. Patients received combination T and MMF therapy at three MMF doses: 1, 1.5, and 2 g/day administered twice daily. A 12-hour blood PK profile for T was obtained prior to MMF dosing; concomitant 12-hour profiles for T, mycophenolic acid (MPA), and mycophenolic acid glucuronide (MPAG) were obtained after 2 weeks of administration. Tolerance was monitored through 3 months. The intra- and intergroup PK of T were variable. The mean AUC0-12 of T for each group was increased after 2 weeks of concomitant MMF administration, but the increase was not statistically significant. Both drugs were well tolerated. Gastrointestinal events were of interest as such have been attributed to both T and MMF. Events reported were diarrhea, nausea, dyspepsia, and vomiting. Other common adverse events were headache, hypomagnesemia, and tremors. Most were mild, although a few were considered to be moderate. There was no apparent relationship between the incidence of any adverse event and MMF treatment group. In the present study, the coadministration of T and MMF did not significantly alter T pharmacokinetics. PMID- 10806607 TI - Effect of metoprolol and verapamil administered separately and concurrently after single doses on liver blood flow and drug disposition. AB - Nine healthy males participated in a double-blind, placebo-controlled, randomized, crossover study to determine the effects of verapamil and metoprolol administered alone and concurrently on blood flow through the hepatic artery and portal and hepatic veins and to detect a possible drug interaction between the two agents. Single oral doses of placebo/placebo, metoprolol (50 mg)/placebo, verapamil (80 mg)/placebo, or verapamil/metoprolol were separated by at least 14 days. Liver blood flow through individual hepatic vessels was measured up to 8 hours after dosage administration using a duplex Doppler ultrasound technique. Cardiac output, heart rate, blood pressure, stroke volume, and total peripheral resistance were measured for 3 hours after drug doses were given. In 5 subjects, pharmacokinetic parameters for total drug as well as S- and R-enantiomers were also measured. Verapamil given alone caused a rapid and intense increase in liver blood flow (hepatic artery = 50%, portal vein = 42%, hepatic vein = 55%) 0.75 to 1 hour after administration because of a decrease in total peripheral resistance and an increase in heart rate, stroke volume, and cardiac output. Metoprolol given alone caused a slow but prolonged decrease in liver blood flow (maximum decrease: hepatic artery = -54%, portal vein = -21%, hepatic vein = -27%) 4 hours after administration because of a decrease in heart rate and cardiac output. When the two agents were given together, a composite of the changes noted after separate administration was noted: a brief peak increase in liver blood flow at 0.33 to 1 hour followed by a slow, prolonged decrease that reached its maximum decline 4 to 5 hours postdose. During the combined phase, metoprolol and its enantiomers had an increased AUC and Cmax, while verapamil and its enantiomers had an increased AUC and t1/2. These pharmacokinetic changes were consistent with the magnitude and time course of liver blood flow changes through the hepatic artery and portal or hepatic veins. PMID- 10806608 TI - Lack of interaction between tolcapone and tolbutamide in healthy volunteers. AB - To assess the effect of tolcapone (an inhibitor of cytochrome P450 [CYP] 2C9 in vitro) on the pharmacokinetics and hypoglycemic effect of the CYP 2C9 substrate tolbutamide, 12 healthy male volunteers were randomized to receive a single dose of tolbutamide 500 mg plus either placebo or tolcapone 200 mg after an overnight fast and 30 minutes after the start of a 6.5-hour 5% glucose infusion (150 mL/h). The participants crossed over to receive the alternative regimen after a washout period of at least 7 days. Tolcapone had no effect on the pharmacokinetics of tolbutamide or its metabolites and did not influence the effect of tolbutamide on plasma glucose concentrations. No serious adverse events or abnormal laboratory results or vital signs were reported. In conclusion, clinically relevant drug drug interactions between tolcapone and tolbutamide when given together in clinical practice appear unlikely. PMID- 10806609 TI - In vitro assays for measuring TGF-beta growth stimulation and inhibition. PMID- 10806610 TI - Measurement of active TGF-beta generated by cultured cells. PMID- 10806611 TI - Iodination of TGF-beta, TGF-beta-receptor crosslinking, and immunoprecipitation of TGF-beta-receptor complexes. PMID- 10806612 TI - Detection of TGF-beta in body fluids and tissues. PMID- 10806613 TI - Analysis of TGF-beta-mediated synthesis of extracellular matrix components. PMID- 10806614 TI - Characterization of Smad phosphorylation and Smad-receptor interaction. PMID- 10806615 TI - Promoter analysis of TGF-beta responsive genes by transient transfection and deletional/mutational analysis. PMID- 10806616 TI - Regulation of AP-1 activity by TGF-beta. PMID- 10806617 TI - Cdk pathway: cyclin-dependent kinases and cyclin-dependent kinase inhibitors. PMID- 10806618 TI - Activation of the mitogen-activated protein kinase pathway by transforming growth factor-beta. PMID- 10806619 TI - Detection of TGF-beta type II receptor hot-spot mutations: the BAT-RII assay. PMID- 10806620 TI - Mutation detection in the TGF-beta receptors and smad genes: RT-PCR and sequencing. PMID- 10806621 TI - Analysis of TGF-beta-inducible apoptosis. PMID- 10806622 TI - [Functional MRI: its present and future]. PMID- 10806623 TI - [Gamma knife radiosurgery for skull base meningiomas: the treatment results and patient satisfaction expressed in answers to a questionnaire]. AB - Skull base meningiomas can be treated by radiosurgery with low morbidity. We evaluated the effectiveness of gamma knife radiosurgery for skull base meningiomas. We also evaluated patient satisfaction through answers to a questionnaire. We treated 77 patients of skull base meningiomas using gamma knife radiosurgery from January 1994 to June 1998. Among these patients, 73 patients were able to be followed up from 6 to 60 months (average 25 months). The diagnosis was made by operations on 39 patients (53%), and on magnetic resonance imagings in 34 patients (47%). 21 patients (28.7%) had residual tumors after operations and 18 patients (24.3%) had recurrence after operations. 40% of neurological deficits occurred after previous operations and 20% of the patients were asymptomatic. The locations of the tumors were mainly the cavernous sinus region in 35 patients and the petroclival region in 20 patients. The tumor sizes were 8.6 to 58.3 mm (average 24.7 mm) and five cases of tumors with a mean diameter above 40 mm were treated by two-staged radiosurgery. Treatment volume was 0.3 to 31 ml (average 9.8 ml). The treatment dose was 8 to 16 Gy (average 11.2 Gy) at the tumor margin. The tumor decreased in size in 50 patients (68%) and was unchanged in 20 patients (28%) and we were able to achieve 96% tumor growth control. The clinical symptoms improved in 22 patients (30.1%) and were unchanged in 47 patients (64.4%) and only 4 patients (5.5%) showed deterioration. 50 patients (68%) felt clinical improvement and only 3 patients (4%) sensed deterioration. 58 patients (79%) were able to continue their previous work. Gamma knife radiosurgery has a very low morbidity rate and enables good tumor control, and patients with skull base meningiomas express satisfaction with the treatment results. PMID- 10806624 TI - [Radiosurgery for cystic metastatic brain tumor]. AB - Stereotactic radiosurgery (SRS) for brain metastasis results in a high local control rate. But cystic metastatic tumor should have been a contraindication for SRS. Because it is often found that a cyst is too large to be irradiated, the tumor does not exist in the center of the irradiation field. Between 1995 and 1998, 8 consecutive patients underwent linear accelerator-based SRS for cystic brain metastases identified by computed tomography or magnetic resonance image scan. Stereotactic cyst aspiration is carried out after placement of the BRW frame under local anesthesia. All of the patients except one were confirmed to have sufficient reduction of the cysts. 5-7 days after stereotactic cyst aspiration, SRS was performed. The dose range was 25-30 Gy. In follow-up MRI, local recurrences and enlargement of cysts were not noted. Six patients with neurological symptoms recuperated satisfactorily. Median survival was 30 weeks from the date of radiosurgery. All of the patients died and the causes of death were related with the primary lesion. We conclude that our technique, a combination of stereotactic cyst aspiration and SRS is an effective measure which leads to palliation of neurologic symptoms and is a low risk treatment for patients with cystic brain metastasis. PMID- 10806625 TI - [Efficacy of antibiotic prophylaxis in clean neurosurgical operations: a comparison of seven-day versus one-day administration]. AB - We compared seven-day and one-day administration of antibiotics as prophylaxis in clean neurosurgical operations. A total of 175 patients were examined. Seven-day administration (protocol A) was studied retrospectively from May 1997 to May 1998 in 87 patients of 46 craniotomies and 41 burr hole operations. One-day administration (protocol B) was studied prospectively from June 1998 to July 1999 in 88 patients of 39 craniotomies and 49 burr hole operations. Protocol A received 1.5 g of sulbactam/ampicillin = 1:2 (SBT/ABPC) preoperatively, and further doses were continued at 12-hour intervals for seven days after the operation. Protocol B received 1.5 g of SBT/ABPC preoperatively and intraoperative additional antibiotics were administered at 5-hour intervals, and only one single dose was administered after the operation. In both of the protocols, 60 mg of tobramycin was mixed with 500 ml of saline for irrigation of operative fields. No postoperative infection occurred in either of the two protocols. We conclude that one-day administration of prophylactic antibiotics is enough to prevent postoperative infections in clean neurosurgical operations. PMID- 10806626 TI - [Two cases of methicillin-resistant Staphylococcus aureus (MRSA) sepsis following craniotomy]. AB - We report here two cases of MRSA sepsis following craniotomy. In case 1, a petroclival meningioma was subtotally removed and lumbar drainage was inserted postoperatively to prevent cerebrospinal fluid leakage. Ventriculo-peritoneal shunt was performed after meningitis was treated with vancomycin and panipenem/betamipron. Two weeks after the procedure, the patient revealed continuous spiking fevers related to MRSA sepsis, which did not improve with vancomycin and arbekacin administration. The focus of infection was found by scintigraphy and CT by 67Ga to be spondylo-diskitis at the level of L2-L3. The lesion was removed and bone from the iliac crest grafted. In case 2, seven days after surgery for multiple meningioma, the patient exhibited spiking fevers and swelling in the left leg. The central venous catheter was removed from the left femoral vein and MRSA was found from blood culture. The patient was treated with arbekacin (200 mg/day). Venous thrombosis diagnosed by CT was treated with heparin. Symptoms related to the infection and laboratory data did not improve because the concentration of arbekacin in the blood did not reach an effective level. The symptoms markedly improved when the dose of arbekacin was doubled (400 mg/day). PMID- 10806627 TI - [T-cell lymphoproliferative disorder involving the central nervous system in two cases]. AB - Lymphoproliferative disorder (LPD) sometimes occurs in immunosuppressive patients. B-cell proliferation is predominantly seen in patients with LPD and T cell proliferation is rare. We here report two patients with T-cell LPD involving the central nervous system (CNS). A 7-year-old boy developed increased intracranial pressure and computed tomography showed a ring-enhanced mass lesion in the right frontal lobe. A 10-year-old boy developed gait disturbance and magnetic resonance imaging showed a non-enhanced lesion in the left parietal lobe. Radiological findings could not differentiate CNS-LPD from other intracranial diseases. Only histologic examination could diagnose LPD in both cases, and Epstein-Barr virus associated RNA was found in the infiltrated T-cell of the latter case. Although chemotherapy with antitumor agents was effective for remission of LPD in both cases, both patients died 3 years after their operations. As CNS-LPD shows rapid progression and frequently results in fatality, this disease requires immediate histologic diagnosis followed by extensive chemotherapy. PMID- 10806628 TI - [A case of angiographically occult arteriovenous malformation with metaplasia (so called brain stone)]. AB - The patient was a 69-year-old female. Right hemiparesis occurred on April 25, 1996, and then was relieved a day later. Because headache (dull pain in the left) persisted subsequently, she consulted our department on April 26. Head CT showed, without enhancement effect, osseous high density on the surface of the left frontal area. MRI showed high intensity on T1 and low intensity on T2 with flow void like findings. Cerebral angiography showed a pooling of contrast medium in the same region. 123I-IMP-SPECT revealed reduced cerebral blood flow in the left frontal and parietal lobes just under the same region. On June 11, the patient underwent surgery during which a tumor with arachnoid hypertrophy was extracted en block. Histopathologically, there were abnormal blood vessels with elastic fibers, expanding to an ossified site, and AVM accompanying ossification was thus diagnosed. Postoperative 123I-IMP-SPECT showed improved cerebral blood flow in the left frontal and parietal lobes. The patient was discharged on June 22. The TIA pathologic condition, a symptom of its onset, was considered attributable to cerebral blood flow steal due to AVM. PMID- 10806629 TI - [A case of a scalp arteriovenous fistula associated with Rendu-Osler-Weber disease treated by direct percutaneous embolization]. AB - We present a case of Rendu-Osler-Weber disease (hereditary hemorrhagic telangiectasia) accompanied by a scalp arteriovenous fistula, which was successfully treated by direct percutaneous embolization. A 51-year-old man, who had multiple vascular telangiectases and pulmonary arteriovenous fistulae, developed an enlarging pulsatile mass of the scalp anterior to the site of the previous craniotomy for a brain abscess in the right occipital lobe. Angiography demonstrated a high-flow arteriovenous fistula between the right superficial temporal artery and a varix. Percutaneous injection of pure ethyl alcohol was planned but seemed risky because of the major drainage being into the bilateral cavernous sinuses through the superior ophthalmic veins. A 24-gauge plastic needle was placed in the right superficial temporal artery just proximal to the fistula, and 0.7 ml of a mixture consisting of n-butyl cyanoacrylate and lipiodol in a ratio of 1:1 was injected. Then, the varix was directly punctured, and retention of the contrast medium was confirmed under manual compression by the placement of a circular ring. Embolization of the varix using 1.0 ml of 70% glucose solution and a subsequent 1.0 ml of pure ethyl alcohol was performed with compression, resulting in total occlusion of the fistula. The scalp mass resolved gradually and there was no evidence of recanalization. We conclude that direct percutaneous embolization is the first therapeutic choice for a scalp arteriovenous fistula with multiple shuntings associated with Rendu-Osler-Weber disease. Dangerous venous drainage should be eliminated before performing embolization with ethyl alcohol. PMID- 10806630 TI - [Intramedullary spinal cord metastasis associated with hemorrhage: a case report]. AB - A rare case of intramedullary spinal cord metastasis associated with hemorrhage was reported. A 74-year-old man had a subacute onset of paraparesis. He became almost paraplegic within a few days. MRI revealed an intramedullary spinal lesion in the epiconus at the Th 11 and Th 12 level, but spinal angiography did not show any abnormality. Since repeated MRI showed enlargement of the lesion, surgery was performed under the diagnosis of an intramedullary spinal cord tumor. Under general anesthesia, a midline myelotomy of about 3 cm was performed and a grayish, elastic and circumscribed tumor as well as a liquefied hematoma in the caudal part was observed. Both the tumor and the hematoma were removed almost totally. The patient's paraparesis improved slightly after surgery. The histological diagnosis was adenocarcinoma. The primary source was unknown, but multiple small metastatic tumors were found in the lung, liver and brain, etc. Hemorrhage from intramedullary spinal cord metastasis is extremely rare with only 6 reported cases in the recent literature. Rapid deterioration of symptoms caused by the hematoma may make the diagnosis more difficult. Indication of surgical treatment should be carefully determined because prognosis of intramedullary spinal cord metastasis is generally very poor. PMID- 10806631 TI - [Left Sylvian fissure meningioma in a one-year-eight-month old child]. AB - We report a case of a Sylvian fissure meningioma in a one-year-eight-month old child who experienced the onset of a convulsive seizure. He had no neurological deficit and no developmental disorders. Computed tomography (CT) and magnetic resonance imaging (MRI) showed a large left temporal tumor which was well enhanced and without dural attachment. Angiography revealed a slight tumor stain in the left Sylvian fissure supplied by branches of the internal carotid artery. Total removal of the tumor was performed, and we found that the tumor had no dural attachment, but was strongly attached to the M2 segment of the left middle cerebral artery. Pathological examinations revealed it to be a fibrous meningioma without malignancy. This is the youngest case among the reported five pediatric deep Sylvian meningiomas. Introducing this case, we discuss the clinical features of pediatric meningiomas. PMID- 10806632 TI - [Hypertensive putaminal hemorrhage with extensive subarachnoid hemorrhage presenting extravasation of contrast material during angiography: case report]. AB - The authors describe a case of hypertensive putaminal hemorrhage with extensive subarachnoid hemorrhage. On admission, the patient aged 71 presented right-sided motor weakness. CT scan on admission revealed left putaminal hemorrhage with extension into the ipsilateral thalamus and lateral ventricle as well as into the subarachnoid space of the suprasellar, ambient, interhemispheric and contralateral sylvian cisterns. To exclude vascular lesions, left carotid angiography was performed just after admission. The lateral view was unremarkable, but the anterior-posterior view demonstrated extravasation of contrast material from the left lateral lenticulostriate artery. The angiographic sylvian point was shifted to the lateral side. No abnormal vessels were revealed. CT scan after angiography showed exacerbation of both intracerebral and subarachnoid hemorrhages, but the consciousness level was unchanged. CT-guided stereotactic aspiration of the hematoma was performed 4 days after the onset, but failed to remove much hemtoma. The patient died of aspiration pneumonia 9 days after onset. The authors emphasize that extensive subarachnoid hemorrhage in cases with hypertensive putaminal hemorrhage may be an important finding which indicates high risk of rebleeding. PMID- 10806634 TI - Reconsidering antiretroviral therapy in HIV: lessons from San Francisco. PMID- 10806633 TI - [Perineural spreading along the trigeminal nerve in a patient with primary intracranial malignant lymphoma: a case report]. AB - We report a rare primary intracranial malignant lymphoma which spread along the trigeminal nerve through the skull base foramen. The patient was a 50-year-old woman, who was diagnosed as having a primary intracranial malignant lymphoma in the right temporal lobe and had undergone an operation and radiation 5 years previously. The tumor was reduced in size and no recurrent tumor could be detected for 5 years. The patient complained of left face swelling and CT scan revealed a large mass in the pterygopalatine fossa. MRI revealed the recurrent tumor in the left Meckel's cave with extension into the cavernous sinus. The tumor extended through the foramen ovale into the pterygopalatine fossa, through the superior orbital fissure into the orbital cavity and through the infraorbital fossa into the face subcutaneously. Biopsy of the subcutaneous tumor was carried out and the pathological diagnosis was malignant lymphoma, B cell type, which was identical with the initial tumor. MRI revealed the enlarged trigeminal nerve and 3D-CT revealed the enlargement of the infraorbital fossa and the foramen ovale. We suspected that primary intracranial malignant lymphoma had recurred in the left Meckel's cave and the tumor had spread along the peripheral three divisions of the trigeminal nerve. Perineural spreading along the trigeminal nerve passing through the skull base in patients with nasopharyngeal carcinoma is not rare, but this rarely occurs in the case of intracranial tumors. PMID- 10806635 TI - Bacillary angiomatosis and Kaposi's sarcoma in AIDS. PMID- 10806636 TI - Immunologic predictors of HIV-related survival among Texas prison inmates. AB - Research indicates that being incarcerated adversely affects overall health status. Because HIV-infection is a growing problem among the U.S. prison population, understanding how incarceration affects HIV-related survival holds particular clinical and public health relevance. Moreover, while the prognostic roles of CD4 and CD8 lymphocyte count have been well documented in noninstitutionalized populations, little is known about how these factors operate to predict survival among prison populations. The present study examined immunologic determinants of HIV-related survival in a cohort of 752 Texas Department of Criminal Justice (TDCJ) inmates who were treated for HIV/AIDS between 1993 and 1996 at a large south-western medical center. Survival analysis using proportional hazards modeling showed that: (1) the prognostic role of CD4 count among inmates was similar to previous findings among non-incarcerated populations; (2) the prognostic role of CD8 count was slightly weaker than that previously reported for non-incarcerated populations; and (3) inmates who exhibited high levels of both CD4 and CD8 count had a survival advantage over those who had a high score on only one of the two factors. PMID- 10806637 TI - Relationships between perception of engagement with health care provider and demographic characteristics, health status, and adherence to therapeutic regimen in persons with HIV/AIDS. AB - The purpose of this descriptive, correlational study was to examine the relationships between perception of engagement with health care provider and demographic characteristics, health status, and adherence to therapeutic regimen in persons with HIV/AIDS. The convenience sample of 707 non-hospitalized persons receiving health care for HIV/AIDS was recruited from seven U.S. sites. All measures were self-report. Perception of engagement with health care provider was measured by the newly developed Engagement with Health Care Provider scale. Adherence to therapeutic regimen included adherence to medications, provider advice, and appointments. Health status was measured by the Medical Outcomes Study Short Form 36 (MOS SF-36), Living with HIV scale, CD4 count, and length of time known to be HIV-positive. There were no significant relationships between engagement with health care provider and age, gender, ethnicity, and type of health care provider. Subscales of the MOS SF-36 and Living with HIV explained a significant, but modest amount of the variance in engagement. Clients who were more engaged with their health care provider reported greater adherence to medication regimen and provider advice. Clients who missed at least one appointment in the last month or who reported current or past injection drug use were significantly less engaged. PMID- 10806638 TI - Microbicides: forging scientific and political alliances. PMID- 10806639 TI - Mechanisms of mucosal immunity: how does the dendritic cell fit in? PMID- 10806641 TI - Microbicide for prevention of sexually transmitted diseases using a pharmaceutical excipient. AB - Preferred microbicides are expected to inactivate most sexually transmitted viral and nonviral pathogens, including HIV-1, without affecting lactobacilli, components of the natural defense system against sexually transmitted diseases (STDs), be widely available, be inexpensive, and have an established safety record for human use. We show here that cellulose acetate phthalate [C-A-P enteric coating polymer (Eastman)], a compound used for coating of enteric tablets, meets all these criteria. PMID- 10806640 TI - Mechanism of thrombospondin-1 anti-HIV-1 activity. PMID- 10806643 TI - Treatment guidelines on web. PMID- 10806642 TI - HIV/AIDS case histories: acute leukemia in an AIDS patient. PMID- 10806644 TI - NIH starts STI study. PMID- 10806645 TI - Calanolide looks promising. PMID- 10806646 TI - Estrogen may protect against HIV. PMID- 10806647 TI - HAART and bone loss. PMID- 10806648 TI - Nevirapine and perinatal transmission. PMID- 10806649 TI - Viral load and transmission. PMID- 10806650 TI - Less concern about HIV. PMID- 10806651 TI - Dementia trial underway. PMID- 10806652 TI - Oral sex more risky. PMID- 10806653 TI - Long way to go for vaccines. PMID- 10806654 TI - [The changes and significance of the intensity of transcription of endothelial nitric oxide synthase-mRNA in placental tissue from cases of pregnancy induced hypertension]. AB - OBJECTIVE: To determine the cause of decreasd expression of endothelial nitric oxide synthase (eNOS) in placental tissue from cases of pregnancy induced hypertension (PIH). METHODS: 9 placentae of normal pregnant women and 12 placentae of patients with PIH were studied. eNOS-mRNA was measured in placental tissue by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1) There was eNOS-mRNA in placental tissue of normal pregnancy as well as PIH cases. (2). The intensity of transcription of eNOS-mRNA in placental tissue of PIH cases decreased significantly when compared with that of normal pregnancy (P < 0.05). CONCLUSION: The decrease of the intensity of transcription of eNOS-mRNA in placental tissue of PIH patients may result in reduction of expression of eNOS in placental tissue of PIH. PMID- 10806655 TI - [Study on relationship between detection of interleukin-6 and its mRNA and pregnancy induced hypertension]. AB - OBJECTIVE: To investigate the effect of Interleukin-6 (IL-6) on pathogenesis of pregnancy induced hypertension (PIH) and to confirm the derivation of IL-6 and relationship between IL-6 and growth of placenta and fetus. METHODS: Pregnant women without infection and uterine contraction are divided into two groups: (1) PIH group (n = 57), (2) Normal pregnancy group (n = 15), reverse transcription polymerase chain rection (RT-PCR) is used to detect IL-6 mRNA expression in leucocyto of peripheral blood and in placenta; Enzyme-linked immunosorbent assay (ELISA) is used to detect IL-6 level of peripheral blood and amniotic fluid. RESULTS: (1) The level of IL-6 and its mRNA in peripheral blood has a increasing trend as PIH degree. In moderate and severe PIH, IL-6 and its mRNA level are significantly higher than that of normal pregnancy. In each degree of PIH, IL-6 mRNA expression in placeata is significantly lower than that of normal pregnancy. In moderate and severe PIH, IL-6 level in amniotic fluid is significantly lower than that in normal pregnancy (P < 0.001). (2) In moderate and severe PIH, IL-6 level has high correlation to the IL-6 mRNA in peripheral blood (r = 0.67, P < 0.01). The IL-6 level in amniotic fluid has significant correlation to IL-6 mRNA in placenta (r = 0.79, P < 0.01). IL-6 and its mRNA in peripheral blood has no high correlation to IL-6 in amniotic fluid. (3) In severe PIH, the IL-6 mRNA in placenta and IL-6 level in amniotic fluid in the group of pregnancy with intrauterine growth retardation (IUGR) are both lower than that of the group without IUGR (P < 0.05). CONCLUSION: In PIH, the immune activity is rising, the IL-6 is overproduced by activated immune cells and it may involve in immune respond of organism and damage of vasicular endothelium. Decrease of IL-6 mRNA in placenta and IL-6 in amniotic fluid showes that the capability of production and transport of IL-6 decline in placenta of PIH patient, and these result in ischemia of placenta trophoblast and disturbance of the growth and development of placenta and fetus. PMID- 10806656 TI - [The changes of plasma calcitonin gene-related peptide level in women with normal pregnancy and pregnancy induced hypertension]. AB - OBJECTIVE: To study the change of calcitonin gene-related peptide (CGRP) concentrations in maternal plasma of the normal pregnant women and patients with pregnancy induced hypertension (PIH) and relationship with CGRP and pathogenesis of PIH. METHODS: The plasma levels of CGRP in 59 cases of normal pregnant women (normal pregnancy group), 57 cases of PIH (PIH group), and 10 cases of nonpregnant women (control group) were determined with a sensitive radioimmunoassay. RESULTS: Plasma CGRP levels in normal pregnancy group increased with advance for the gestational week, reached the peak value (76.84 +/- 27.75 ng/L) in late trimester; plasma CGRP levels in PIH group decreased with series degree of PIH, levels of CGRP in moderate (49.76 +/- 23.11 ng/L) and severe PIH (46.35 +/- 22.32 ng/L) were significantly lower than that in mild PIH (71.28 +/- 21.98 ng/L) and the normal late pregnancy, the differences were significant (P < 0.005). There was no correlation between CGRP levels and the mean arterial pressure (MAP). CONCLUSIONS: The change of CGRP may play an important role in the pathogenic mechanism of PIH. PMID- 10806657 TI - [The study on in vitro antibody secretion of B lymphocyte in patient with pregnancy induced hypertension]. AB - OBJECTIVE: To study the immunologic etiopathology of pregnancy induced hypertention (PIH) in some extent by analyzing the antibody secretion of B lymphocytes of PIH patients. METHODS: The subjects were divided into two groups: PIH group (n = 15) and normotensive control group (n = 15). Peripheral lymphocytes of the subjucts were separated by Ficoll-Hypaque density centrifugation. The lymphocytes were cultured in RPMI-1640 medium with or without pokeweed mitogen (PWM) for ten days. Then the supernatant was collected and the IgG and IgM were measured by enzyme linked immunosorbent assay (ELISA). The results of the two groups were analyzed by using student test. RESULTS: Both IgG and IgM secretion of B lymphocytes between PIH group and normtensive control group showed no difference in the absence of PWM (per 1 x 10(6) lymphocytes, for IgG, 120 +/- 68 ng vs 154 +/- 89 ng, P > 0.05; for IgM, 337 +/- 119 ng vs 371 +/- 135 ng, P > 0.05. Whereas the level of the two antibodies in PIH group was significantly decreased than that of the normotensive group in presence of PWM (for IgG, 947 +/- 518 ng vs 1,367 +/- 598 ng, P < 0.05; for IgM, 2,023 +/- 879 ng vs 2,840 +/- 1,057 ng, P < 0.05). CONCLUSION: B lymphocyte of PIH patient has a decreased capacity of antibodies secreting in response to stimulation of PWM, which suggests that B lymphocyte of PIH patient may have functional deficiency. PMID- 10806658 TI - [Serum concentration of insulin-like growth factor-I in cord blood]. AB - OBJECTIVE: To evaluate the role of insulin-like growth factor-I (IGF-I) in fetal growth and placental growth. METHODS: Cord blood IGF-I was determined by radioimmunoassay (RIA) after serum samples were extracted with acid-ethanol. Linear correlation between variables was tested by calculating pearson's correlation coefficient. student's test was used to compare mean values between groups. RESULTS: In 102 tested samples, the concentration of cord blood serum IGF I was correlated with the gestational age, the birth weight of neonates and placental weight (r = 0.32, P < 0.001, r = 0.68, P < 0.001, r = 0.75, P < 0.001). The mean serum level of IGF-I in the small-for-gestational-age (SGA) neonates was significantly lower than that in average-for gestational-age (AGA) (t = 4.77, P < 0.001), But the serum concentration of IGF-I in the Large-for-gestational-aga (LGA) neonates was significantly higher than that of AGA neonates (t = 7.97, P < 0.001). CONCLUSION: Those results suggested that IGF-I is an important regulator for fetal and placental growth and development. It has the key role for macrosomia and IUGR forming. PMID- 10806659 TI - [Effect of the long chain polyunsaturated fatty acids levels in mothers on fetuses]. AB - OBJECTIVE: To investigate the effect of the Long Chain Polyunsaturated fatty acids (LCP) levels in mothers on fetuses. METHODS: The plasma and red blood cell membrane LCP contents were measured and compared between 48 mothers and 48 fetuses. RESULTS: The plasma concentration of linoleic acid (LA) and linolenic acid (ALA) in mothers was higher than that in fetuses, arachidonic acid (AA) and docosahex aenoic acid (DHA) were lower. The positive relations were observed between the maternal and the cord plasma concentration of LA, ALA, AA and DHA, AA content in maternal was related to birth weight. CONCLUSION: Maternal nutrient and placental function are important for maintaining fetal growth and development. PMID- 10806660 TI - [The changes of serum estrogen, progesterone and the function of immune system in intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To explore the effects of estrogen, progesterone and the function of immune system on intrahepatic cholestasis of pregnancy (ICP). METHODS: Fifty women with ICP and fifty normal pregnant women were enrolled in the study. The levels of their serum estradiol (E2) and progesterone (P) were measured and cellular immunity and humoral immunity were determined. Radioimmunoassay was used to detect the levels of E2 and P, and APAAP was used to detect cellular immunity. RESULTS: (1) The level of estrodiol was significantly increased in ICP group than those of normal control group (25.89 +/- 6.85 micrograms/L, 16.92 +/- 4.98 micrograms/L, P < 0.01). (2) The level of CD8+ was decreased (19.06 +/- 1.93%, 26.43 +/- 2.89%) and the ratio of CD4+/CD8+ was increased significantly in ICP group than those of control group (2.23 +/- 0.38, 1.73 +/- 0.23, P < 0.05). (3) There were negative correlation between E2 and CD8+, and positive correlation between E2 and the ratio of CD4+/CD8+. CONCLUSION: These results suggest that the high level of E2 of ICP patients may result in functional disorder of immune system by the means of estrogen receptor on CD8+. This may be a factor of ICP development. PMID- 10806661 TI - [Factors affecting the clinical pregnancy rate in an in vitro fertilization and embryo transfer program]. AB - OBJECTIVE: To analyze the various factors in an in vitro fertilization and embryo transfer (IVF-ET) program which may affect the clinical pregnacy rate. METHODS: A retrospective study was done on 559 IVF-ET cycles from 1992-Nov. 1995. The indication for treatment was bilateral tubal blockage. The chi 2 analysis of single factor variants with SPSS-PC + V3.0 was used for statistics. RESULTS: The overall clinical pregnancy rate in 559 cycles was 21.6%. The cause of tubal blockage due to tuberculoses consisted of 28.4%, and 34.9% of secondary sterility had the history of artificial abortion. The changes of environment, the different causes of tubal blockage, the history of previous intrauterine pregnancy did not affect the clinical pregnancy rate. When the number of embryos transferred increased to 5, the clinical pregnancy rate was highest 32.5%. The cumulative embryo score or embryo quality was related significantly with clinical pregnancy rate. CONCLUSIONS: The number and quality of embryos transferred are important factors affecting the clinical pregnancy rate. However, measures to prevent high order multiple pregnancy and studies on the survival potential of embryos besides their morphology should be emphasized. PMID- 10806662 TI - [Role of hyperinsulinemia in pathogenesis of polycystic ovary syndrome and treatment by reduction of insulin secretion]. AB - OBJECTIVE: To investigate the role of insulin in the polycystic ovary syndrome (PCOS) pathogenesis and application of Metformin in the treatment of PCOS. METHODS: Serum androgen, luteinizing hormone (LH) and sex hormone-binding globulin(SHBG) concentration during fasting and serum 17 alpha hydroxyprogesterone (17-OHP), LH level in response to gonadotropin-releasing hormone agonist (GnRH-a) stimulation were determined. Oral glucose-tolerance tests(OGTT) before and after oral administration of metoformin for 8-12 weeks were performed in 12 obese and 11 lean women with PCOS. RESULTS: After oral administration of metformin for 8-12 weeks, fasting insulin concentration in obese group and area under curve (AUC) after OGTT in lean group decreased significantly. There were significant decrease in basal 17 alpha hydroprogesterone, androstenedione, testosterone concentration, and increase in serum SHBG concentration in obese and lean groups. Basal LH and the response of serum 17-OHP, LH to GnRHa were not significantly changed. CONCLUSIONS: Hyperinsulinemia may play an important role in hyperandrogenism GnRH-a of PCOS, and metformin may be used in the treatment of PCOS. PMID- 10806663 TI - [Determination of cellular and humoral immunity in mice of autoimmune ovarian failure]. AB - OBJECTIVE: To determine cellular and humoral immunity in mice of autoimmune ovarian failure. METHODS: A mice model of autoimmune ovarian failure was established. The percentages of help T cell (CD4+) and suppressor T cell (CD8+), the ratio of CD4/CD8 and anti-ovarian antibody (AOA) were determined. RESULTS: No significant changes were found in CD4+ fraction, but the percentage of CD8+ was reduced and the ratio of CD4/CD8 increased. AOA was positive which was probably the cause of damage of the ovarian tissues, reduction of sex hormone secretion and impaired fertility. CONCLUSION: Autoimmune ovarian failure is correlated with the abnormal change of CD4/CD8 and the increase of B-lymphocytes function. PMID- 10806664 TI - [Roles of prostaglandins and leukotrienes in the pathogenesis of amniotic fluid embolism]. PMID- 10806665 TI - [Study of magnesium and calcium levels of plasma and within erythrocyte before and after magnesium sulfate treatment in patients with pregnancy induced hypertension]. AB - OBJECTIVE: To study the changes of calcium and magnesium levels in patients with pregnancy induced hypertension (PIH) and the therapeutic effect of magnesium sulfate (MgSO4). METHODS: The calcium and magnesium levels in plasma and erythrocytes of 36 patients with PIH and 14 healthy gravidi in their late pregnancy were determined and the changes after 4 hours infusion of MgSO4 as well. RESULT: Plasma calcium decreased significantly in moderate and severe PIH patients, but calcium increased and magnesium decreased in their erythrocytes. After MgSO4 infusion for PIH, magnesium levels in both plasma and erythrocytes increased, while calcium increased in plasma and decreased in erythrocyte to normal levels. CONCLUSION: Magnesium deficiency is the basis of calcium and magnesium disturbances in PIH, and MgSO4 treatment may improve the situation effectively. PMID- 10806666 TI - [Study of interleukin-6 and tumor necrosis factor-alpha levels in maternal serum and amniotic fluid in patients with preterm premature rupture of membrane]. AB - OBJECTIVE: To study the change and clinical significance of maternal serum and amniotic fluid interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) levels in patients with chorioamnionitis and premature rupture of membrane (PROM). METHODS: Twenty normal term-pregnant women as control, and fourty-six women with PROM were enrolled for study. Maternal serum and amniotic fluid IL-6, TNF-alpha levels were measured by radioimmunoassay and enzyme-labeled immunosorbent assay (ELISA). Chorioamnionitis was diagnosed histologically. RESULTS: The maternal serum IL-6 levels and amniotic fluid IL-6, TNF-alpha levels were higher in PROM group than that of control (P < 0.01). There was significant positive correlation between maternal IL-6, amniotic fluid IL-6, TNF-alpha levels and the duration from rupture of membrane, (the longer was time, the higher were the maternal IL-6 and the amniotic fluid IL-6. TNF-alpha). There were 12 cases with chorioamnionitis in patients with PROM and their maternal and amnotic fluid IL-6, TNF-alpha levels were higher than that of non-chorioamnionitis (P < 0.01 0.05). CONCLUSION: IL-6 and TNF-alpha levels in maternal serum and amniotic fluid are valuable clinical indices for identification of chorioamnionitis in patients with PROM. PMID- 10806667 TI - [The role of diet consultation on improvement of the diet nutrition in pregnant women]. AB - OBJECTIVE: To study the improvement of the intakes of various nutrients including protein, fat, carbohydrate, fibers, calcium, iron, zinc et al, in pregnant women after appropriate diet consultation by doctors. METHODS: 100 cases of pregnant women coming to our diet consultation clinic were randomly selected, in which 50 cases (control group) came for the first time and another 50 cases (study group) were for the third visit at least. The diet nutrition of the study group after the second consultation were compared with that before consultation and with concurrent control group. RESULTS: After diet consultation, except for retinol, the intake of various nutrients, including calorie intake, protein, fibers, magnesium, calcium, phosphorus, iron, zinc, vitamin E, riboflavin, thiamine, nicotinic acid, in study group were significantly improved compared with that of the study group before diet consultation and of the control group (P < 0.01). CONCLUSION: Diet consultation plays excellent role in promoting scientific and appropriate nutrients intake of pregnant women. PMID- 10806668 TI - [Prognostic value of basal hormone levels on ovarian reserve]. AB - OBJECTIVE: To study the prognostic value of basal hormone levels on ovarian reserve during ovulation induction. METHODS: Day 2 serum follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) levels of 112 ovulation induction cycles (60 infertile patients) were measured by radioimmunoassay. The relationship between basal hormone levels and numbers of dominant follicles and pregnancy rates were analysed. RESULTS: Day 2 serum FSH > or = 15 IU/L predicted a poor response to ovarian stimulation, with fewer dominant follicles and lower pregnancy rate, so was day 2 serum E2 > or = 45 ng/L. Day 2 serum LH level was unrelated to pregnancy rate. CONCLUSION: Day 2 FSH and E2 levels are useful in prognostic assessment of ovarian reserve. PMID- 10806669 TI - [Analysis of characteristics of genital diseases in girlhood]. AB - OBJECTIVE: To explore the characteristics of genital diseases in girlhood in order to improve its prevention and treatment. METHODS: 106 girls with genital diseases (age < or = 12 years) admitted to Obstetrics and Gynecology Hospital of Shanghai Medical University from Jan. 1977 to Dec. 1996 were reviewed retrospectively. They were divided into two group. 1st group 36 cases (1977 1986), 2nd group 70 cases (1987-1996). The clinical data of these two groups were compared and also compared with the whole patients admitted to the Gynecological Department of this hospital at the same period. RESULTS: The patients' number of the second group significantly increased by 0.1 percentage as compared with the 1st group (P < 0.001). The characteristics of genital diseases in the second group is different from that in the 1st group: the incidence of genital tumor ranked the highest, the age of genital malformation patients became younger, and the inflammatory disease tended to be more complicated and serious. CONCLUSIONS: These data indicated that the pediatrician and gynecologist should pay more attention to the prophylaxis and treatment of genital diseases in girlhood. PMID- 10806670 TI - [The effect of estrogen replacement therapy on plasma nitric oxide levels in postmenopausal women]. AB - OBJECTIVE: To investigate the effect of estrogen replacement therapy (ERT) on plasma nitric oxide (NO) levels in postmenopausal women (PMW). METHODS: The plasma concentrations of NO and serum concentrations of estradiol (E2) were assessed before and 4 weeks after placebo or ERT administration in 10 cases of healthy PMW (control group), 17 cases of healthy PMW (healthy group) and 21 cases of PMW with hyperlipidemia (hyperlipidemia group) respectively. RESULTS: There was no difference in NO concentrations between before and after placebo administration in control group. In healthy and hyperlipidemia groups, the mean concentrations of NO were significantly higher 4 weeks after ERT administration than baseline. The positive correlation between the mean percent increase of NO and the E2 concentrations after ERT administration was statistically significant in healthy group. CONCLUSION: The increase of plasma NO concentrations caused by ERT may contribute to it cardiovascular protective effect in PMW. PMID- 10806671 TI - [Laparoscopic assisted transvaginal hysterectomy]. AB - OBJECTIVE: To evaluate the indication, technical procedure and the clinical value of laparoscopic assisted trans vaginal hysterectomy (LAVH). METHODS: Ninety-eight patients suffered from various kinds of benign gynecological diseases underwent LAVH from Feb. 1995 to Feb. 1997. The mean age was 52 years (range 38-66), preoperative examination findings were 38 cases of normal uterine size, 60 of uterine enlargement (44 less than 12 week gestational size and 16 larger). Among these patients, 29 cases had a history of lower abdominal laparotomy. RESULTS: 2 out of the 98 cases were switched to laparotomy (2.0%). Mean operation time of LAVH was 106 min (range 60-240 min). The amount of blood loss was approximately 50-150 ml during operation, and the average hospitalization was 6 days. CONCLUSION: Laparoscopic assisted vaginal hysterectomy expands the indication of transvaginal hysterectomy, and avoids laparotomy and lessens injuries, shortens the hospitalization. It is a promising approach of hysterectomy and should be used widely. PMID- 10806672 TI - [Randomized comparative study of GyneFix IN and TCu 380A intrauterine devices]. AB - OBJECTIVE: To observe the clinical performances of the new intrauterine device (IUD)-GyneFix IN. METHODS: The present study is a randomized comparative clinical trial. 607 healthy parous women were randomly allocated into GyneFix IN group (n = 302) or TCu 380A group (n = 305). IUD was inserted during the menstrual interval by the trained investigators. Follow-up were arranged at 1, 3, 6 and 12 months after insertion. The discontinuation rates were calculated by life table method. RESULTS: At the end of the first year, there was no pregnancy occurred in GyneFix IN group. Its expulsion rate and removal rate for medical reasons were 2.67 and 1.02 per 100 women respectively. The use-related discontinuation rates was 3.66, which was significantly lower than that in the TCu 380A group (7.88, P < 0.05). The number of women with complaint of pain was also less in GyneFix IN group. CONCLUSION: The excellent performance of the new IUD which is a frameless device with high copper surface and anchoring system was confirmed by this multicentre trial. Due to lower expulsion rate and less side effect of pain, it could be recommended. PMID- 10806673 TI - [Effects of mifepristone on the serum nitric oxide level and uterine blood flow in early pregnant women]. AB - OBJECTIVE: To study the effects of mifepristone on serum nitric oxide (NO) level and uterine blood flow in early pregnant women. METHODS: Serum NO concentrations of 58 early pregnant women and 15 normal nonpregnant women were measured by a copper-coated cadmium reduction method, and uterine artery and trophoblastic blood flow were determined by color doppler flow imaging (CDFI). The ultrastructure of villi and decidua were studied under electron microscopy. RESULTS: Compared with nonpregnant women, serum NO concentration was significantly higher in early pregnant women(P < 0.01). After administration of mifepristone, serum NO level was significantly decreased, resistance index(RI) and systolic to diastolic ratio(S/D) ratio of uterine artery and trophoblastic blood flow were significantly higher than that of normal early pregnancy(P < 0.05). CONCLUSION: Mifepristone may exert effects on uterine blood flow by decreasing serum NO level. PMID- 10806674 TI - [The clinical and pathological characteristics of exaggerated placental site]. AB - OBJECTIVES: To study the pathological and clinical features of exaggerated placental site (EPS) and to propose its diagnostic criteria and differential diagnosis. METHODS: The pathological features, immunohistochemical marks, clinical characteristics and prognosis of 13 cases with EPS were studied and analyzed. RESULTS: The histological feature of EPS was that trophoblasts mainly intermediate trophoblast (IT), infitrated into decidua and myometrium layers, while original histological structure was not destroyed, and placental site features remained partially. The immunohistochemical marks of EPS, HPL and EMA, were positive or strongly positive. The primary clinical presentation of EPS was postpartum hemorrhage and the prognosis was good in 5 years. CONCLUSIONS: EPS is a kind of gestational trophoblastic disease primarily involving IT. It may be a preneoplastic lesion of placental site trophoblastic tumor (PSTT). Its differential diagnosis depend on combining histological, immunohistochemical findings, and clinical features. Attentions should be paid to EPS in cases with postpartum hemorrhage. PMID- 10806675 TI - [Anti-tumor effect of cytotoxic T lymphocytes induced by soluble antigen of ovarian carcinoma and anti-CD3 monoclonal antibody on ovarian carcinoma]. AB - OBJECTIVE: To evaluate the therapeutic effect of the new method--adoptive cell immunotherapy on ovarian carcinoma. METHODS: Tumor soluble antigen (TSA) was extracted from cells of ovarian carcinoma (COC1 and COC2n). To generate cytotoxic T lymphocytes (CTL), peripheral blood mononuclear cells (PBMC) collected from healthy persons were induced by TSA and anti-CD3 monoclonal antibody (CD3-McAb). The antitumor effect of CTL on COC1 cells in vitro was compared with LAK and CD3 AK, whereas the growth inhibitory effect of CTL on COC2n transplantation tumors in nude mice was compared with CD3-AK. RESULTS: The cytotoxicity of CTL, CD3-AK and LAK was 79.4%, 52.1% and 51.7% respectively (P < 0.01). Mean volume of tumor in nude mice on the 9th day after ovarian carcinoma cells transplanted was 44.4 +/- 24.2 mm3 in CTL group, 118.8 +/- 40.0 mm3 in CD3-AK group and 443.0 +/- 158.7 mm3 in control (no treatment) (P < 0.01), whereas mean duration of survival was 28.5 days, 25.5 days and 17 days respectively (P < 0.01). CONCLUSION: The results indicated that CTL exhibited higher cytotoxic effect in vitro and greater inhibitory effect on tumor growth resulting in longer survival in vivo. It suggested that the adoptive cell immunotherapy might provide a new idea for treatment of ovarian carcinoma. PMID- 10806676 TI - [The effect of carboplatin combined chemotherapy on renal function in patients with ovarian carcinoma]. AB - OBJECTIVE: To investigate the changes of renal function during carboplatin (CBDCA) combined chemotherapy in patients with ovarian carcinoma. METHODS: 26 courses of postoperative systemic CBDCA therapy were carried out in 7 patients with ovarian carcinoma. During chemotherapy both systolic maximum and diastolic minimum blood flow velocity (Vmax, Vmin) and resistance index (RI) in renal segmental artery (SA) and interlobular artery (ILA) were meaured by color doppler weekly. Meanwhile renal creatinine clearance rate (Ccr) were also determined. 14 healthy women were taken as controls. RESULTS: From the 1st week to the 3rd week at the begining of CBDCA therapy, Vmax and Vmin in SA and ILA were significantly decreased (P < 0.01). A significant positive correlation was found between Ccr and the values of Vmin in SA and ILA (r = 0.52, P < 0.01). CONCLUSIONS: The results indicated that carboplatin combined chemotherapy could induce disturbances of renal function. The changes of renal function were related to renal blood flow velocity which could be used as a monitoring index of renal function. PMID- 10806677 TI - [Progress in the basic research of endometriosis]. PMID- 10806678 TI - [P-glycoprotein and multidrug resistance]. PMID- 10806679 TI - [The relationship of insulin resistance and the pathogenesis and perinatal outcome of pregnancy induced hypertension]. AB - OBJECTIVE: To study the insulin resistance in the pathogenesis of pregnancy induced hypertension (PIH), and its relationship to perinatal outcome. METHODS: In 111 PIH and 155 control group, the concentration of C peptide and insulin of maternal blood samples before and after 32 gestational weeks, and fetal blood samples just after delivery was determined. Fetal outcomes were assessed by the neonatal weight at delivery, Apgar scores and the quality and quantity of amniotic fluid. RESULTS: The maternal concentration of C peptide and insulin in PIH was higher than that of control group (P < 0.05, or P < 0.01). In PIH group, the maternal concentration of C peptide and insulin in the subgroup of adverse perinatal outcomes tended to be higher than that in the subgroup of good perinatal outcomes; but in control group, there was no significantly difference between these two subgroups. CONCLUSIONS: Insulin resistance may be one of the factors which couse PIH, and it has significant relationship with advese perinatal outcomes. PMID- 10806680 TI - [Analysis of 196 cases of trial of labor with borderline oligohydramnios assessed by ultrasound]. AB - OBJECTIVE: To determine the effect of borderline oligohydramnios on perinatal outcome. METHOD: Patients admitted for delivery had an amniotic fluid index (AFI) assessment. Color Doppler was used to examine the systolic/diastolic ratio of the blood flow velocity in fetal umbilical arteries. Normal amniotic fluid volume was defined as an AFI of 8.1-18 cm (n = 200), and borderline oligohydramnios as 5.1 8.0 cm (n = 196). The two groups were compared on perinatal outcomes as all of the patients had trial of labor. RESULTS: There was no difference in the incidence of fetal distress and neonatal mortality between the two groups. The incidence of emergency section in borderline oligohydramnios group was significantly higher. CONCLUSION: The patients with borderline oligohydramnios may have a trial of labor under proper intensive care. PMID- 10806681 TI - [Transvaginal ultrasound assessment of cervical changes before onset of labor]. AB - OBJECTIVE: To study the clinical value of cervical measurement by transvaginal ultrasound after 36 weeks of gestation. METHODS: Eighty three normal nulliparous woman were studied from 36 weeks of gestation until delivery prospectivelly. Transvaginal ultrasound examinations were performed and cervical length (CL), cervical width (CW) and the diameter of cervical canal were measured weekly. 52 cases were delivered naturelly and 31 cases by cesarean sections (CS). RESULTS: From 36 weeks of gestation until delivery, there was a significant correlation between gestation age (GA) and CL and the diameter of cervical canal (r = -0.37, P < 0.00001; r = 0.16, P = 0.002). The time from examination to initiation of labor was significantly correlated with CL and the diameter of cervical canal (r = 0.32, P < 0.00001; r = -0.19, P = 0.02). The regressive formulas were CL (cm) = 13.77 - 0.27 x GA, the time to beginning of labor (week) = 0.64 + 0.54 x CL (cm). The cases with CL > or = 3.5 cm at 2-3 weeks before labor had a significant higher CS rate than those with CL < or = 3.5 cm (P = 0.002). CONCLUSIONS: Transvaginal ultrasonographic measurement of CL, CW and the diameter of cervical canal could reflect the maturity of cervix accurately. It could be used as predictors of labor course and outcome of delivery. PMID- 10806682 TI - [Perforin protein and gene expression in the different phases of endometrium]. AB - OBJECTIVE: To study the expression of perforin protein and gene in the different phases of endometrium and the role of perforin in the reproductive system. METHODS: Immunohistochemical and in situ hybridization techniques were employed to demonstrate the population, phenotype of the perforin positive cell in endometrum of different phases of menstrual cycle, first-trimester decidua and postmenopausal endometrium. RESULTS: Perforin positive cells were found to be located in the endometrial stroma and expressed natural killer-like cell surface marker (CD56 antigen, also named NK-like cell or CD56+ lymphocyte). The quantitative analysis of perforin protein cells demonstrated an increase in number from proliferative endometrium to secretory endometrium through first trimester decidua (P < 0.05). These cells were absent in the postmenopausal endometrium. Perforin mRNA positive cells formed islands in proliferative endometrium stroma, but the perforin protein expression cells were scattered. There was difference between the perforin gene expression and protein expression in proliferative endometrium. In first-trimester decidua, perforin mRNA positive cells incresed. Some perforin mRNA positive cells located around spiral arterioles were observed. CONCLUSIONS: Expression of perforin in the different phases of endometrium is specific and plays an important role in the reproductive function. They probably exert some influence on embryo implantation and may be involved in endometrial stromal breakdown during menstruation. PMID- 10806683 TI - [The study of endothelin and endothelin receptors in cyclic human endometrium]. AB - OBJECTIVE: To determine the distribution of endothelin (ET) peptide and endothelin as well as their receptors (ETaR and ETbR) mRNA in human cycling endometrium in order to assess possible role of ET in endometrial reproductive physiology. METHODS: Twenty-three endometrial specimens from gynecological surgery for benign conditions were studied by means of immunohistochemistry, in situ hybridization and computer quantitative image analysis. RESULTS: The expression of ET peptide and ET mRNA in human endometrium varies throughout the menstrual cycle, increasing gradually from proliferative phase to secretory phase, with maximal levels in the pre-menstrual phase. Stromal expression is lower than in glandular epithelium and remains relatively constant in the menstrual cycle. ETaR and ETbR mRNA are detected in normal human endometrium, higher in glandular epithelial cells than in stromal cells. ETaR mRNA expression has no obvious cyclic changes, but ETbR mRNA expressions are higher in secretory endometrium than in proliferative endometrium. The ratio of ETaR/ETbR has also cyclic changes. CONCLUSION: Changes of Expression of ET and ET receptors in the endometrium throughout the menstrual cycle suggest that ET might be important in endometrial regeneration and repair following menstruation, and probably plays a role as the paracrine control of the uterine vascular bed. PMID- 10806684 TI - [Comparative study of the third party and paternal leukocyte immunization in recurrent spontaneous abortion of lowered maternal-fetal immuno-recognition]. AB - OBJECTIVE: To compare curative effect of leukocyte immunization to recurrent spontaneous aborters(RSA) with lowered maternal-fetal immuno-recognition between transfusions of the third party leukocyte-rich erythrocyte concentrates and injections of paternal leukocytes. METHODS: Forty RSA patients with lack of blocking antibodies were randomly divided into two groups. Blocking antibodies were analysed by one-way mixed lymphocyte reaction blocking test and complement dependent cytotoxic test, anti-idiotypic antibodies to blocking antibodies by one way mixed lymphocyte reaction blocking inhibiting test, and flow cytometry evaluation of blocking antibodies were used to investigate effect of blocking antibodies on the CD antigens of T cells. The immunization was achieved by transfusions of the third party leukocytes or injections of the paternal leukocytes for 3 times or more at an interval of 4 weeks. RESULTS: It was found in dynamical analyses that levels of the blocking antibodies and their anti idiotypic antibodies increased significantly after the aborters had been immunized. The successful pregnancy rate was 86.2% following immunization, and body weight of the infants was 3,300 +/- 116 g(x +/- s), with no significant difference to 3,279.0 +/- 113.8 g of the infants from normal pregnancy. According to dynamics of blocking antibodies and their anti-idiotypic antibodies, the paternal leukocyte immunotherapy appeared superior to the third party one, but the successful pregnancy rate and the body weight of infants were with not different between the two immunotherapy groups. CONCLUSION: The third party and paternal leukocyte immunization could effectively stimulate the aborters of lowered maternal-fetal immuno-recognition to produce blocking antibodies and their anti-idiotypic antibodies, leading to successful pregnancy. The immunotherapy does not apparently influence the body weight of the delivered infants. PMID- 10806685 TI - [Ovarian follicular maldevelopment in infertile women]. AB - OBJECTIVE: To study the manifestation and outcome of ovarian follicular maldevelopment (FM) in infertile women. METHODS: Among 296 infertile women, 80 cases with FM were taken as study group and 20 with normal ovulation as control. Serial B-ultrasonography, cervical mucus scoring, serum estradiol (E2) assay and laparoscopy, endometrium histology were monitored in both groups. RESULTS: The incidence of FM in infertile women was 27.0%, B-ultrasonography showed many small and obscure outlined follicles in the ovarian. The maximum and mean cervical mucus socres in FM group were < 9 and 7.3 +/- 1.8, significantly lower than those in control (> 10, 13.2 +/- 1.8) (P < 0.01). So were the midcycle serum E2 levels (300 +/- 100 pmol/L vs 900 +/- 400 pmol/L, P < 0.001). No stigma was found in two third of FM patients and no corpus luteum in about one-third. Proliferative endometrium was shown in 34.0% of the FM group. CONCLUSION: (1) FM with an incidence of 27.0% might be an important cause of infertility. (2) FM results finally in follicular atresia or multiple immature luteinized follicles. PMID- 10806686 TI - [Improvement of mouse early embryo development in vitro by co-culture with human oviductal epithelial cells in serum-free medium]. AB - OBJECTIVE: To examine the effect of human oviductal epithelial cells on early mouse embryonic cleavage and growth in vitro. METHODS: Ninety 2-cell mouse embryos were co-cultured with human oviductal epithelial cells in DMEM/Ham's F12 + 0.3% bovine serum albumin(BSA) + estradiol (E2) and ninety embryos cultured in DMEM/F12 + 0.3% BSA + E2 alone as control. RESULTS: Among the embryos co-cultured with oviductal epithelial cells, 82% developed to the morulae stage, 77% cavitated to blastocysts with 63% hatching, as compared with 45% to morulae stage, 13% to blastocysts and none hatching in the controls. Co-cultured embryos cleaved significantly faster than control and showed no or less fragmentation than those in the control. CONCLUSION: These results suggested that human oviductal epithelial cells can support early embryonic development and yield a reasonable number of embryos with good quality up to the blastocyst stage. PMID- 10806687 TI - [The comparison of biological characteristics between tumor-infiltrating lymphocytes and tumor-associated lymphocytes in ascites of epithelial ovarian carcinoma]. AB - OBJECTIVE: To compare biological charicteristics between tumor-infiltrating lymphocytes (TIL) and tumor-associated lymphocytes (TAL) in epithelial ovarian carcinomas. METHODS: The surgical samples of four ovarian specimens and eight cancer ascites from epithelial ovarian carcinoma were collected, and ascites were divided into bloody and non-bloody groups. The cancer tissue was dissected to yield single cells suspensions. TIL and TAL were grown in coculture with autologous tumor cells and 500 U/ml recombinant interleukin-2 (rIL-2). Phenotype was studied using flow cytometry for a varity of human immunocompetent cell surface markers (CD3, CD4, CD8, CD25). Cytotoxicity was investigated using 4-hr 51Cr-release assays with primary ovarian carcinoma cell cultures and the CAOV3 cell line and the K562 cell line as target cells in cultured period. RESULTS: (1) There was no significant difference in CD3, CD4, CD8 and CD25 expression among the three groups (P > 0.05). CD4 to CD8 (CD4/CD8) ratio was consistently more than 1 in cultured period. (2) In all three groups, Natural killer (NK) acitivity was high and no difference among them (P > 0.05). (3) In all twelve cases, TIL and TAL in the non-bloody ascites possess higher potential to lyse the autologous tumor target than TAL in the bloody ascites (P < 0.01) when effector cell to target cell (E:T) was 50 to 1 (50:1). Lytic activity was 30.2%-51.0% and 10.8% 15.8% respectively. CONCLUSIONS: TIL and TAL in non-bloody ascites in epithelial ovarian cancer have the same biological characteristics. PMID- 10806688 TI - [The comparison of radiosensitivity between uterine cervical squamous carcinoma and adenocarcinoma]. AB - OBJECTIVE: To compare the radiosensitivity of uterine cervical squamous carcinoma and adenocarcinoma. METHODS: The shrinkage of tumor and the radiation-induced damage were compared between cervical squamous carcinoma and adenocarcinoma after twice afterloading radiotherapy. Proliferating cell nuclear antigen index (PI), detected immunohistochemically, was compared before and after radiotherapy between the two tumors. RESULTS: Patients of squamous carcinoma with tumor shrinkage 50% or more accounted for 70% (21/30) and patients of adenocarcinoma accounted for 27% (4/15), which was statistically significant (P < 0.01). The radiation-induced damage grade II b-III accounted for 60% (18/30) in squamous carcinoma and for 20% (3/15) in adenocarcinoma, which was statistically significant (P < 0.05); PI decreased from 53.50% +/- 4.34% to 39.30% +/- 4.02% after radiotherapy in squamous carcinoma (P < 0.001), while PI decreased from 35.47% +/- 3.83% to 31.80% +/- 3.06% in adenocarcinoma (P > 0.05). CONCLUSIONS: The radiosensitivity of cervical squamous carcinoma is higher than that of adenocarcinoma. The decrease of PI after radiotherapy may be a biological basis of radiosensitivity for uterine cervical squamous carcinoma. PMID- 10806689 TI - [Interleukin 2 gene transfer on the proliferation and morphology of ovarian cancer cell line SKOV3]. AB - OBJECTIVE: To study the effect of interleukin-2 (IL-2) gene transfer on the proliferation and morphology of ovarian cancer cell line SKOV3. METHOD: IL-2 gene was introduced into ovarian cancer cell line SKOV3. The morphologic changes, mitosis, proliferation curve, 3H-uptaking and cell cycle were studied. RESULTS: IL-2 cDNA, 490 bp, was transduced. IL-2 gene expression was detected. IL-2 activity was 30-318 U/10(6) cells/24hr. IL-2 gene modified cell is squamas like with smaller nucleus/cytoplasm ratio, larger cell area and more homogeneous. The ultrastructural study showed that the cytoplasma of IL-2 gene modified cells is markedly vacuolized and the chromatin is condensed. There was less mitosis found in SKOV3/IL-2 cells. SKOV3/IL-2 cell proliferate more slowly, the doubling time prolonged from 38.40 hr to 65.36 hr, 3H-uptaking decreased 52.3% of the parent cell. The cell cycle study showed that SKOV3/IL-2 cell was blocked to phase G1/G0. CONCLUSIONS: The retrovirous vector can transduce Il-2 gene into ovarian cancer cell line SKOV3 effectly. IL-2 gene transfer can induce morphologic changes of SKOV3 cells, which may indicate the biologic behavior changes of the targets. IL-2 gene transfer can induce the proliferation inhibition of SKOV3 cells. PMID- 10806690 TI - [Clinical study on pelvic venous congestion syndrome after tubal sterilization]. AB - OBJECTIVE: To study the clinical features, diagnosis and therapy of pelvic venous congestion syndrome after tubal sterilization. METHODS: By using laparoscopy combined with transmyometrial pelvic venophogram and samples after operation, pathomorphology of pelvic congestion venous is observed. RESULTS: The coincidence rate of laparoscopic diagnosis is 94.6% (53/56), compared with transmyometrial pelvic venophogram. There is no significant difference for light or severe type patients (P > 0.05). The cure rate of the cases in this group is 60.7%, the improvement rate is 16.1%, total effectiveness rate is 76.8%. The effectiveness rate of operational treatment is 90.1%, conservative treatment is 18.2%. CONCLUSIONS: Laparoscopy is very valuable for the diagnosis of this disease. Laparoscopy combined with transmyometrial pelvic venophogram can be regarded as the major method to diagnosis pelvic venous congestion syndrome. It is recommended that operation for patients with medium and severe symptoms. PMID- 10806691 TI - [Genetic study of pre-eclampsia]. PMID- 10806693 TI - [Proper using drugs in obstetrics and safety of mother and fetus]. PMID- 10806692 TI - [Effects of chemotherapy on the changes of histology and function of the ovary]. PMID- 10806694 TI - [Summary of national congress on proper using drugs in obstetrics]. PMID- 10806695 TI - [The effect of dolantin used during labor on the fetal heart rate and uterine contraction]. AB - OBJECTIVE: To investigate the effect of dolantin on fetus and uterine contraction. METHODS: Thirty pregnant women under stress, restless or fatigue were given dolantin 100 mg i.m. during the latent phase of labor. Internal fetal heart rate (FHR) monitoring and intrauterine pressure measuring were carried out at 30 minutes, 1 hour and 2 hours after medication. RESULTS: The baseline FHR showed a decrease 30 minutes after dolantin, and decreased further at 1 and 2 hours though still within normal range. At the same time, the baseline of intrauterine pressure decreased as well at 30 minutes, so hypertonic uterine contractions were improved. One hour later the incoordinate uterine contraction was relieved and the frequency and strength of uterine contraction were enhanced (P < 0.01). CONCLUSION: During labor, especially in latent phase, the use of dolantin may improve the general condition of the women. In spite of the decrease of FHR, no fetal distress occurred. Besides, it may enhance the frequency and strength of uterine contraction and relieve the incoordinate uterine contraction. PMID- 10806696 TI - [The effect of nimodipine on retinal blood flow in pregnancy induced hypertension]. AB - OBJECTIVE: To assess the effects of nimodipine on retinal blood flow in patients with pregnancy induced hypertension (PIH). METHODS: Fourteen patients were studied with color flow Doppler before and after 30 mg nimodipine administered orally, while other fourteen patients received intravenous magnesium sulfate as control group. Central retinal artery flow velocity waveforms were measured and pulse index (PI) were calculated. RESULTS: Nimodipine reduced the central retinal artery PI significantly from 0.92 +/- 0.14 to 0.75 +/- 0.22. It also reduced systolic blood pressure from 20.7 +/- 2.5 kPa to 19.8 +/- 1.5 kPa and diastolic blood pressure from 14.4 +/- 2.3 kPa to 13.6 +/- 1.9 kPa. CONCLUSIONS: Nimodipine may dilate central retinal arteries. Changes in pulsatility index in retinal arteries maybe indicative of similar changes in other cerebral vessels. Nimodipine may be another choice of PIH therapy. PMID- 10806697 TI - [The clinical study on labor pain relief using two kinds of segmental block anesthesia]. AB - OBJECTIVE: To study the pain relief effectiveness of combined subarachnoid peridural segmental block and simple peridural anesthesia, and their influences on the mothers and the infants. METHODS: 100 pregnants women were administered combined subarachnoid and peridural segmental block during labor and delivery (study group). 40 pregnant women received simple peridural anesthesia (control group). The degree of labour pain, side effects, labor course, the mode of delivery and the incidences of postpartum hemorrhage, fetal distress and neonatal asphyxia were observed in two groups respectively. RESULTS: The pain relief effect in the study group was much better than that of the control group (P < 0.01). There were no significant differences of the mean time of labor course and the mode of delivery between the two groups (P > 0.05). The incidences of postpartum hemorrhage, fetal distress and neonatal asphyxia were similar (P > 0.05). CONCLUSION: The analgesic delivery of combined subarachnoid and peridural segmental block is safe and effective, which has no influences on the labour course and the mothers and infants. Its pain relief effectiveness is more positive and satisfactory than that of simple peridural anesthesia. We suggest that it should be performed in the medical units under optimal conditions. PMID- 10806698 TI - [Clinical study on reduction of postpartum bleeding in cesarean section by misoprostol]. AB - OBJECTIVE: To observe the effects of misoprostol on reduction of postpartum bleeding in cesarcan section. METHODS: One hundred and eighty-two cesarean sections were randomized into three groups: misoprostol group (n = 60), 600 micrograms misoprostol was given orally when peritoneum was incised; misoprostol and oxytocin group (n = 64), 600 micrograms misoprostol was given orally and 20 IU oxytocin was injected into uterine muscle immediately after delivery of baby. Oxytocin group (n = 58), 20 IU oxytocin was injected into uterine muscle and 20 IU intravenous injection immediately after delivery of baby. The amount of bleeding within 2 hours after delivery was measured. RESULTS: The mean amount of bleeding in misoprostol group was 212 +/- 56.0 ml, in misoprostol and oxytocin group was 208 +/- 55.4 ml, oxytocin group was 345 +/- 64.7 ml, respectively. The difference was significant P < 0.01 between misoprostol group and oxytocin group, there was no differences between misoprostol group and misoprostol plus oxytocin group. CONCLUSION: Misoprostol is more effective in reduction of postpartum bleeding than that of oxytocin. PMID- 10806699 TI - [Antenatal treatment of maternal-fetal Rh incompatibility hemolysis disease]. AB - OBJECTIVE: To study the effective antenatal treatment of maternal-fetal Rh incompatibility hemolysis disease to improve fetal outcome. METHODS: Besides therapy of plasmapheresis, the immune globulin intravenous infusion (IVIg) was added for antenatal treatment of fetal hemolysis disease due to Rh incompatibility. Every 7-20 days 10 g IVIg was administered according to the titer of Rh antibody in maternal serum. RESULTS: 5 Rh negative pregnant women with poor obsterical history were treated with this protocol. Total IVIg infusion was 12 times with an average of 2.4 times per case, and there was no side effects. Plasmapheresis was used 18 times totally with an average of 3.6 per case. There was no fetal death. All the five newborns underwent exchange transfusion and IVIg therapy, and all were survival. CONCLUSIONS: IVIg may enter fetal blood circulation via placenta, and adjust fetal immune ability to reduce the degree of fetal hemolysis. PMID- 10806700 TI - [An analysis of antenatal psychological status and its related factors]. AB - OBJECTIVE: To provide basis for antenatal psychological counselling by investigating psychological status and related factors of parturients. METHODS: Analysing psychological status of 100 primigravida by self-made questionnaires. Symptom checklist go (Scl-go), HAMD and HAMA. RESULTS: Single-factor analysis showed: six factors, that were social behavior, mood controlling, history of abortion, relation with parents, psychological preparation for delivery and family roles, are correlated with psychological status of pregnant women. Multi factor analysis indicated: 3 factors i.e. relation with parents, psychological preparation for delivery and social behavior were positively correlated with psychological status of parturients. CONCLUSIONS: Relation with parents, preparation for delivery and social behavior are the most important factors to influence antenatal psychological status and useful references for antenatal psychological counselling. PMID- 10806701 TI - [Clinical analysis of intracranial venous embolism during pregnancy and puerperium]. AB - OBJECTIVE: To study the cause and clinical manifestation of intracranial venous embolim during pregnancy and puerperium, and its early diagnosis and treatment. METHODS: 18 cases of intracranial venous embolism confirmed by Computed Tomography (CT) were eligible for this retrospective study. RESULTS: Of all the cases, 8 cases occurred before delivery and 10 post partum; 10 cases complicated by pregnancy induced hypertension and 8 cases postpartum hemorrhage. Headache occurred most frequently, followed by coma and seizure. Blood rheologic assay showed hypercoagulable states in 6 cases. For 11 cases treated with urokinase, 10 were cured, while in the remaining patients, 3 of 6 cured. CONCLUSIONS: The results show that hypercoagulability states were related to intracranial venous embolism. Coma and seizure were the main characters. CT was an important diagnostic measure. Lowering intracranial pressure, anticoagulation, dethrombosis and timely ending pregnancy were the main treatment measures. The early use of urokinase may exert better effect. PMID- 10806702 TI - [Placental morphometriological study on prolonged and delayed pregnancy and its relationship to pregnancy outcome]. AB - OBJECTIVE: To study the morphological characteristics of the placental villous vessels and uteroplacental spiral arteries in the prolonged and delayed pregnancy and its relationship to pregnancy outcome. METHODS: Placentae from 20 cases of prolonged pregnancy (PP), 15 cases of delayed pregnancy (DP), and 29 cases of normal term pregnancy (NTP) were examined by morphometriological feature image analysis. RESULTS: Significant differences were found between PP and NTP groups (P < 0.05) in the following findings: the placental weights, the numbers of the placental villus with the syncytial knots, cytotrophoblasts, thickening of basal lamina, fibrinoid necrosis, vasculosyncytial membranes, and the diameter, perimeter and area of the villi the spiral artery wall thickening of the uteroplacental bed, and the incidences of the oligoamnios and amniotic fluid meconium staining. Significant differences of the villus perimeter and oligoamnios were only found between DP and NTP (all P < 0.05). There was no significant differences in the indices between PP and DP. As the pregnancy delayed and prolonged, the placental weights gradually decreased and the low Apgar score of neonates gradually increased. CONCLUSION: After term pregnancy the placental function is gradually lowered especially in PP. The situations of the placenta, amniotic fluid, and fetus should be frequently monitored after 40 gestational weeks. PMID- 10806703 TI - [Apoptosis of human oocytes is age-related]. AB - OBJECTIVE: To study the morphological characteristics, and rate of apoptosis of human oocytes in relation to age. METHODS: 206 oocytes were aspirated from antral follicles 4-6 mm in diameter from 58 patients who had surgery done for ovarian cysts. These oocytes were incubated in Ham's F10 with 10% fetal cord serum (FCS) for 48-120 hours, and examined under inverted microscope every 6-8 hours to see their morphological changes. In situ end-labeling (ISEL) of fragmented DNA was used to confirm apoptosis. RESULTS: The characteristics of apoptosis are: (1) shape change of oocytes. (2) oocyte shrinkage. (3) chromatic condensation and apoptotic body formation. (4) internucleosomal DNA cleavage was shown by ISEL. Apoptosis occurred in 13.9% (women aged 21-30), 37.7% (women aged 31-40), and 61.5% (women aged 41-50) respectively. The rate of apoptosis was much higher in women aged 41-50 than that in the other two groups. CONCLUSION: The rate of apoptosis of human immature oocytes cultured in vitro is much higher in older women aged 41-50 than in those aged 21-40, which may account for the poor response to superovulation stimulation and low oocyte quality in older women. PMID- 10806704 TI - [Experimental study of apoptosis induced by cisplatin in human ovarian carcinoma AO 10/17 cells]. AB - OBJECTIVE: Whether apoptosis induced by cisplatin resulted in tumor growth inhibition and the relationships between apoptosis and cell cycle, intracellular and extracellular Ca++ concentration were investi- gated. METHODS: AO 10/17 cells were treated with cisplatin. Morphological changes (HE stain) were observed under light microscope; DNA degradation was assayed by electrophoresis on agarose gel. The change of cell cycle was analyzed by flow cytometer and cytosolic Ca++ concentrations were monitored using probe of calcium ion fluo-3/AM. RESULTS: Under the influence of cisplatin, AO 10/17 cells exhibited changes of apotosis both in morphology and in the characteristics of electrophoresis. Meanwhile the cell cycle also showed special change and intracellular Ca++ concentration was increased. Increase in extracellular Ca++ concentration when AO 10/17 cells were treated with CaCl2 and cisplatin did not enhance the efficacy of cisplatin. CONCLUSION: The results suggest that apoptosis followed by change of cytosolic Ca++ concentration is one of the mechanism of cisplatin in inhibition of ovarian tumor growth. It is closely related to cell cycle. PMID- 10806705 TI - [Clear cell carcinoma of vagina: report of 3 cases and review of literatures]. AB - OBJECTIVE: To summarize the clinical characteristics and treatment of vaginal clear cell carcinoma. METHODS: 3 cases of vaginal clear cell carcinoma admitted from 1958-1996 were analyzed retrospectively. RESULTS: Neither intrauterine diethylstilbestrol exposure nor diethylstibestrol use during their life was found in 3 cases. After external pelvic radiation and intracavitary 137Cs radiotherapy, 5-year survival was achieved in all 3 cases. 2 of them were alive with tumor free for 16 years. CONCLUSION: The results suggested that the prognosis of vaginal clear cell carcinoma was rather good and it could be cured by proper radiotherapy. PMID- 10806706 TI - [Ultrasonography in the surveillance of the uterine cervix during pregnancy]. PMID- 10806707 TI - [Advances in the diagnosis and treatment of gestational trophoblastic disease]. PMID- 10806708 TI - [Uses of computers in obstetrics and gynecology]. PMID- 10806709 TI - [Summary of National congress on computer application in obstetrics and gynecology]. PMID- 10806710 TI - [Computerized transformation of the cardiotocographic paper record to its digital equivalent for computerised analysis]. AB - OBJECTIVE: To analyze cardiotocograph by computer, the tracings recorded in paper form must first be converted into their digital equivalent. We developed a method by which this process may be performed. METHODS: Paper recordings were first scanned using a conventional flat bed scanner to obtain a digital image. Each image was firstly corrected for rotational misalignment error during scanning and, sceondly the grid was removed by performing logistic contrast enhancement to leave the discrete fetal heart rate and tocographic tracings. The method was validated by comparing differences between the fetal heart rate obtained from the paper record with that directly obtained from the fetal monitor. RESULTS: Forty recordings were analyzed. The mean difference per recording between the actual and derived values ranged from -0.26(-)-1.26 beats per minute. The 95% confidence interval for the pooled differences between the derived and actual fetal heart rate values was--7-5 beats per minute. CONCLUSION: By using the techniques described in this paper, it is now possible to convert the large number of paper records available so that they can be analyzed by computerized cardiotocograph interpreters. PMID- 10806711 TI - [Clinical applications of computer system for diet consultation of pregnant women]. AB - OBJECTIVE: To investigate nutritional status of women in different gestational ages and seasons and the role of diet consultation by a computer system on the improvement of intakes of nutrients of pregnant women. METHODS: 712 women in different gestational ages and 747 cases in different seasons received diet consultation by the computer system. A comparison was done for nutritional status between pregnant women with consultation and the women without consultation. RESULTS: Major pregnant women in various gestational ages and seasons was deficient in nutrient intake. However, the nutritional status was significantly improved in women following diet consultation in comparison with the women not received diet consultation. CONCLUSION: Computer system really plays an excellent role in promoting diet consultation and improvement of diet condition of pregnant women. PMID- 10806712 TI - [A computerized information system for gynecological patients]. AB - OBJECTIVE: Clinical experiences in the fields of general gynecology, gynecological oncology and reproductive endocrinology had been accumulated over a long period of time. All of the recorded information was written on various kinds of sheets that could easily be damaged and lost with frequent reading. Statistical works were done manually. This is the situation may encountered by all of the hospitals in China, so a computerized information system for gynecological patients is eagerly needed. METHODS: The program was designed on the basis of Foxpro 2.6 for Windows. RESULTS: The interfaces were specially designed for every of 72 databases to be like the style of patient's records which can be easily operated with mouse pressing buttons. Some of the databases such as basal body temperature, metastasis of lymph nodes and classification of endometriosis were carefully designed. Because the program will be used in China, the interfaces were all in Chinese. A dictionary was used to simplify the importing of the data. When import, one can easily press the line with the appropriate meaning in the dictionary. A strong statistical function was attached to the program. With its friendly interface, statistical work can be done with ease. This system is capable of further development and fit into needs of various hospitals and departments. CONCLUSION: This program is very useful for clinical doctors as well as scientific research personnel. With the help of this program, efficacy of our medical work can be raised, clinical experiences can be summarized in time and the clinical level can thus be improved. PMID- 10806713 TI - [Doppler echocardiographic studies on fetal cardiac function of diabetic mothers]. AB - OBJECTIVE: To assess fetal cardiac function by Doppler echocardiography in diabetic pregnancies. METHODS: We investigated the changes of cardiac function in 54 normal fetuses and 23 fetuses of diabetic mothers. Cardiac anatomic datas were obtained by measuring the diameter of the aortic (AO), the pulmonary artery (PA), the mitral valve(MV), the tricuspid valve (TV) and interventricular septal thickness (IVS). Blood flow evlocity were obtained from the AO, PA, atrioventricular valves. The following variables were measured: peak flow velocity (PFV), accelerate time (ACT), systolic volume (SV) and ForceF. These indices respresented the systolic function. The peak early flow velocity (PFVe), peak late flow velocity (PFVa), PFVe/PFVa(E/A), mean velocity (V) of MV or TV respresented the diastolic function. RESULTS: The results revealed that in fetuses of diabetic mothers, IVS became thickness. The diameter of AO, PA, MV, TV were wider than that of normal fetases. The PFV, SV, ForceF of AO or PA increased significantly. There was no different in ACT between two groups. The PFVa of MV or TV increased significantly, whereas the E/A of both atrioventricular valves decreased. CONCLUSION: It showed that the fetal hyperinsulinemia triggered by maternal hyperglycemia result in fetal weight increasing, as well as hyperplasia and hypertrophy of myocardial cells. An iacreased contractility is compatible with relatively larger size of such fetuses. The intracardiac flow volume increased secondary to the more needs of fetal development. Abnormal thickenning IVS might impair ventricular diastolic function. PMID- 10806714 TI - [Levels of endothelin and beta-human chorionic gonadotropin and their relationship in patients with pregnancy induced hypertention]. AB - OBJECTIVE: To measure levels of endothelin (ET) and beta-human chorionic gonodotropin (beta-hCG) and analyse the correlation between them in patients with pregnancy induced hypertension (PIH), as well as to investigate their role in the pathogenesis of PIH. METHODS: 32 patients with PIH were matched with 17 normal term singleton pregnant women. ET and beta-hCG levels of above patients and 14 nonpregnant health reproductive age women were determined by radioimmunoassay. RESULTS: ET and beta-hCG levels were significantly higher in patients with PIH (30.35 +/- 14.52 ng/L and 7.35 +/- 4.86 nmol/L) than that of two control groups (P < 0.05), especially in middle and severe PIH (P < 0.05), but no significant different in mild PIH (P > 0.05), compared with normal pregnant group (20.79 +/- 7.08 ng/L and 4.13 +/- 1.91 nmol/L) and nonpregnant group (19.78 +/- 6.66 ng/L and 0 nmol/L). There was no significant difference of plasma ET levels between normal pregnant and nonpregnant women (P > 0.05). beta-hCG in nonpregnant woman hadn't been determined. We did not find any association between the levels of ET and beta-hCG in normal pregnancy (r = +0.2216, P > 0.05), but significant positive correlation in the patients with PIH (r = +0.5418, P < 0.01). CONCLUSION: The increased levels of ET and beta-hCG, as well as their positive correlation, suggested that the dysfunction of the placental cells may be associated with the endothelial cell damage in PIH patients. PMID- 10806715 TI - [The relationship between epidermal growth factor and its receptor and fetal birth weight]. AB - OBJECTIVE: To investigate the relationship of epidermal growth factor (EGF) and its receptor (EGFR) with fetal birth weight. METHODS: Using enzyme linked immunoabsorbent assay (ELISA) method the EGF concentrations of materal sera, cord blood sera and amniotic fluids were determined in 40 cases with IUGR, 40 cases with term normal birth weight, 25 cases with fetal macrosomias and 15 cases with normal non-pregnant women. Simultaneously using the immunohistochemical method EGFR in placentas and fetal membrance was determined in all groups of pregnancy. RESULTS: The EGF levels in normal non-pregnant women were lower than that in all groups of pregnancy. The EGF concentrations of materal sera, cord blood sera and amniotic fluid in IUGR group were lower than that in normal birth weight group. EGF levels were no difference between fetal macrosomias group and normal birth weight group. There is a positive correlation between the fetal birth weight and EGF concentrations in maternal sera, cord blood sera and amniotic fluids. In IUGR group the number of EGFR in placent and fetal membrane was less than that in normal birth weight group. In macrosomia group placental EGFR number was more than that in normal birth weight group. CONCLUSIONS: EGF and its receptor levels are related with IUGR, EGF concentrations in maternal sera and amniotic fluids in the third trimester may be a valuable index for assessing the fetal growth. PMID- 10806716 TI - [The relation between human fetal growth and the blood levels of insulin-like growth factor-I]. AB - OBJECTIVE: To determine the relation between human fetal growth and the levels of insulin growth factor-I (IGF-I) in both the maternal and fetal compartments. METHODS: Serum samples were obtained from pregnant woman(n = 171) and their neonate cord blood(n = 164) between 32-42 weeks of gestation. The woman and the neonate were divided into three groups by the birth weight of newborn. In large for gestational age (LGA) group, the 77 cases were from pregnant woman and the 64 cases were from their neonate; in average for gestational age (AGA) group, the 59 cases were from pregnant woman, the 59 cases were from their neonate; and in small for gestational age (SGA) group, the 35 cases were from pregnant woman and 43 cases were from their neonate. The serum samples were analyzed by radioimmunoassay for the levels of IGF-I. RESULTS: The levels of IGF-I in the neonatal serum were lower than those in maternal blood (P < 0.01). The levels of IGF-I in SGA group were lower than those in the other groups (P < 0.05). There was significant correlation between gestational ages and the levels of IGF-I (r = 0.91-0.98), but not in SGA group (r = 0.21-0.81). There was a direct correlation between the levels of IGF-I in the cord blood and the neonatal weight (P < 0.05). CONCLUSIONS: These data demonstrate that the level of expression and regulation of IGF-I in maternal and fetal compartments were independent of each other. The IGF-I level in cord blood is shown to be adjunctively and directly associated with fetal size in pregnancies and the IGF-I plays the precise role in deviant fetal growth. PMID- 10806717 TI - [Localization and quantitative analysis of insulin-like growth factor-I in placenta of extreme fetus]. AB - OBJECTIVE: To investigate the immunohistochemical localization of insulin-like growth factor-I (IGF-I) in human placenta and to evaluate the role of IGF-I in fetal growth and placental growth. METHODS: In 72 tested samples, The cellular distribution of IGF-I in human placenta was determined by using immunohistochemistry. The relative area and mean absorbance were determined by using image analysis technique. Student's t-test was used to compare mean values between groups. RESULTS: IGF-I immunostaining was found in syncytiotrophoblast, cytotrophoblast and Hoffbauer's cells of chorionic villi and chorion of fetal membranes. The strongest immunoreactivity was observed in the syncytiotrophoblast and cytotrophoblast. The relative area and absorbance in placenta of SGA group were significantly lower than those of AGA group respectively (t = 4.65, P < 0.001; t = 2.15, P < 0.05). But the relative arca and absorbance of LGA were significantly higher than those of AGA (t = 8.72, P < 0.001; t = 7.82, P < 0.001). CONCLUSION: These results suggested that IGF-I in human placenta was principally synthsized and secreted by syncytiotrophoblast, cytotrophoblast, hoffbauer's cells of chorionic villi and chorion of fetal membranes. IGF-I is an improtant regulator of fetal and placental growth and development. It has the key role for macrosomia and IUGR forming. PMID- 10806718 TI - [Detection of intrauterine fetal growth of twins]. AB - OBJECTIVE: To investigate how to detect intrauterine fetal growth of twins with height of uterine fundus. METHODS: This study was a randomized coupling trial. 75 couples of twins and single gestation were involved. Linear regression analysis was made after correcting the uterine height of single gestation. RESULTS: If each of twins develops as the same as single gestation, the relationship in uterine height between twins and single gestation can be described as a correlation equation: y = 1.26x + 1.3 (R2 = 0.656, r = 0.81, P < 0.001). The regression coefficient is 1.26, which is very closed to 3 square root of 2. According to the correlation equation, we made a table to forecast the uterine height of twins and single gestation in different gestational age. CONCLUSION: The ralationship in the uterine height between twins and single gestation can be used to predict intrauterine fetal growth of twins. PMID- 10806719 TI - [Mutation of breast cancer susceptibility gene in ovarian cancer and its clinical significance]. AB - OBJECTIVE: To detect breast cancer susceptibility gene (BRCA1) mutation in ovarian cancer and to look for correlations between BRCA1 mutation and hereditary ovarian cancer. METHODS: Mutation of BRCA1 gene in 4 patients with hereditary ovarian cancer and 31 patients with sporadic ovarian cancer were screened by polymerase chain reaction-single strand conformation polymorphism analysis with non-isotopic silver staining method. RESULTS: 2 mutations of BRCA1 gene were found in 2 of 3 patients belonged to hereditary breast-ovarian cancer syndrom (HBOC) which were located in exon 2 and 21 respectively. No mutation was found in 31 cases of sporadic ovarian cancer and 1 case of hereditary site-specific ovarian cancer. CONCLUSION: BRCA1 mutation was probably closely related to hereditary breast-ovarian cancer syndrome. Detection of BRCA1 gene mutation was helpful to diagnose HBOC families. PMID- 10806720 TI - [Uses of artificial intelligence, expert systems and computer-assisted diagnosis and therapy in the field of obstetrics and gynecology]. PMID- 10806721 TI - [Clinical uses of computer-assisted cytological screening system in obstetrics and gynecology]. PMID- 10806722 TI - [Nitric oxide synthesis and its role during pregnancy]. PMID- 10806723 TI - [Development of human preantral follicles and oocytes maturation in vitro]. AB - OBJECTIVE: To study the development of human preantral follicles and oocytes maturation by in vitro culture. METHODS: Preantral follicles obtained from follicular fluid during egg retrieval in an in vitro fertilization (IVF) program was first incubated in Ham's F10 with 15% fetal cord serum (FCS). After 28 days, the medium was supplemented with different doses of hMG, mature follicular fluid (mFF) and epidermal growth factor (EGF) by orthogonal design. Induction of final maturation was completed in the presence of human menopasual gonadotropin hMG and mFF. RESULTS: Development from preantral to antral follicles was found within 6 12 days of culture. With increasing time, the proportion of follicles with diameters of more than 300 micrograms increased at 21-28 days of culture (P < 0.005). The maximal number of oocytes extrusion and first polar body formation occurred in the presence of 1501U/L hMG, 40% (v/v) mFF and 6 micrograms/L EGF. CONCLUSIONS: Follicular fluid obtained during egg retrieval in an IVF program was found to contain many preantral follicles which could develop into antral follicles with extrusion of oocytes in culture, and the oocytes matured in vitro. However, the course of development of follicles and maturation of oocytes in vitro is quite different from that in vivo. PMID- 10806724 TI - [Development of human fetal preantral follicles in vitro]. AB - OBJECTIVE: To study oocyte maturation of human fetal preantral follicles in vitro, the effects of different factors on oocyte maturation. METHODS: Preantral follicles were obtained from fetal ovaries from termination of 20-28 weeks gestation. Factors involved in the development, maturation of fetal preantral follicles were studied in tissue and cell culture by means of Northern blot, and RT-PCR (nested). RESULTS: (1) Preantral follicles obtained from fetal ovary increased in size to more than 120 microns in diameter without antrum formation and with increasing time the preantral oocytes became mature with extrusion of 1st polar body, insulin-like growth factor 1 (IGF-1) had a more marked effect on oocyte maturation, as compared with epidermal growth factor (EGF). (2) Expression of alpha fetoprotein mRNA occurred in fetal follicles on day 5, 15, 25, 35 of culture, and became negative on day 45 of culture. (3) Medium consisted of 1. human menopasual gonadotropin (hMG), 2. follicular fluid, 3. IGF-1, 4. EGF increased the expression of bcl-2 mRNA by 17.6%, 22.4%, 47.1% and 37.5% respectively in fetal follicular cells, and down-regulated the expression of p53 mRNA by 19.8%, 22.1%, 42.6% and 31.6% respectively. CONCLUSIONS: (1) The development and maturation of fetal preantral follicles in vitro differed from those in vivo. (2) Expression of bcl-2 mRNA increased and expression of wild-type p53 mRNA decreased during follicular development and maturation. PMID- 10806725 TI - [Clinical study on relieving labor pain used combined spinal and epidural anesthesia]. AB - OBJECTIVE: To investigate the effect of combined spinal and epidural anesthesia (CSEA) on relieving and eliminating labor pain and the effect on newborns, labor process and delivery mode. METHODS: 80 cases without any obstetric complications and anesthetic contraindications were selected for the study group. CSEA was applied when cervix dilated for about 2-3 cm, 80 pregnant women with similar obstetric condition as control group without any anesthesia. We compared the labor pain, duration, delivery mode and fetal status of two groups. RESULTS: In study group CSEA showed good effect on relief of labor pain, shortening the active phase of labor course and acceleration of cervical dilation. There are no different in the incidence of fetal distress, neonatal asphyxia and postpartum hemorrhage between two groups. CONCLUSION: Apllication of CSEA is useful in relieving labor pain, accelerating labor course, lowering cesarean section rate without any side effect on fetus. PMID- 10806726 TI - [Expression of endothelial nitric oxide synthase gene in placental villus of preeclampsia]. AB - OBJECTIVE: To investigate the expression of endothelia nitric oxide synthase (eNOS) mRNA in the villus of preeclampsia and the relationship between eNOS and preeclampsia. METHODS: Dot blot, using DIG-labeled human eNOS cDNA probe, had been used to examine the expression of eNOS gene in placental tissues of 9 normotensive pregnancy and 9 preeclampsia women. By using Leica QWIN image disposal and analysis system Mean OD of every dot was assessed. RESULTS: The level of eNOS mRNA in placental villus was significantly lower in preeclampsia woman than that of control group (P < 0.001). CONCLUSION: The decrease of eNOS in placental villus of preeclampsia may be one of the causes for human placental dysfunction and high impedance of fetoplacental circulation in preeclampsia. It may play an improtant role in the genesis of pregnancy induced hypertension. PMID- 10806727 TI - [The effect of enzyme-linked immunosorbant assay and polymerase chain reaction on the detection of maternal and fetal human cytomegalovirus infection]. AB - OBJECTIVE: To evaluate polymerase chain reaction (PCR) and enzyme-linked immunosorbant assay (ELISA) on detection human cytomegalovirus (HCMV) infection in preghnant women and fetuses. METHODS: ELISA was used to screen for serum HCMV IgM in 1881 pregnant women. Blood HCMV DNA was examined by PCR in 656 of them simultaneously. 99 fetuses or newborns of HCMV infected mothers and 47 of uninfected ones were detected by both methods. RESULTS: The positive rate of serum HCMV-IgM in pregnant women was 2.4%, and that of HCMV DNA was 12.0%, 17.2% of fetuses or newborns from infected mothers had either positive HCMV-IgM or positive HCMV DNA or both of them. The intrauterine transmission rate of HCMV in mothers with positive HCMV-IgM or positive HCMV DNA or both of them was 18.5%, 14.8% and 27.3% respectively. Simultaneous detection with ELISA and PCR in 656 pregnant women and 146 fetuses showed fine correlations and significant differences between two methods. PCR had significantly higher detection rates than that of ELISA. CONCLUSIONS: ELISA combined with PCR may raise the detection rate and diagnostic efficacy of HCMV infection in both pregnant women and fetuses. PMID- 10806728 TI - [Prediction of cephalopelvic disproportion by ultrasonographic cephalopelic]. AB - OBJECTIVE: To develop a prospective antepartum method of identifying cephalopelvic disproportion by comparing the diameters of fetal head with those of the maternal midpelvis. METHODS: Transvaginal ultrasound pelvimetry was performed on 190 healthy primigravidas with cephalic presentation at 28-35 weeks of gestation, and the diameters of their fetal heads were meassured within one week prior to delivery. These indices the cephalopelvic indices of diameter, cirumference and area, were calculated and compared. RESULTS: The cephalopelvic index of diameter (CID), defined as the difference between the mean diameter of the midpelvis and the fetal biparetal diaameter (BPD), showed the highest degree of accuracy (77.9%). Eighty three percent of women with CID less than 15.8 needed operative delivery; 76.2% of those with CId more than 15.8 mm underwent vaginal delivery. CONCLUSIONS: Transvaginal ultrasound pelvimetry and the CID by and large seems to be able to identify cephalopelvic disproportion before labor and may help obsetricians choose the most appropriate form of delivery in an uncomplicated vertex presentation. PMID- 10806729 TI - [Ultrasonographic diagnosis of peripartum cardiomyopathy]. AB - OBJECTIVE: Analysing the characteristics of ultrasonographic manifestation for peripartum cardiomyopathy (PPCM), and discussing its diagnostic criteria. METHOD: Echocardiographic examination in 20 PPCM and 20 normal volunteers (NC) was completed with Acuson 128 computed sonography. RESULTS: All 20 PPCM showed an enlargement in the left ventricle and left atrium. Left ventricular end diastolic anterio-posterior diameter was 65.4 +/- 4.5 mm, left atrium end systolic anterio posterior diameter was 43.1 +/- 6.1 mm. There was a very significant difference in left atrium and ventricle size between PPCM and NC (P < 0.001). Among 20 PPCM, there were enlargements of the right atrium in 6 cases, of the right ventricle in 8 cases and 20% cardiomegaly. All patients showed decreased left ventricular wall movement. Ejection fraction (0.33 +/- 0.08) and fraction shorting (16.2 +/- 5.0%) were significant different with that of NC (P < 0.001). The regurgitation rate in the heart valvae was MR 100%, TR 70%, PR 70% and AR 15% respectively. CONCLUSION: The ultrasonographic manifestation of PPCM was not specific, so the diagnosis of PPCM depended on peripartum history, left atrium and ventricle enlargements or cardiomegaly, decreased left ventricular function based on ruling out of other heart diseases. PMID- 10806730 TI - [The source of endogenous digitalis-like substance in normal pregnancy]. AB - OBJECTIVE: To study the source of endogenous digitialis-like substance (EDLS) which increases in maternal vein plasma during normal pregnancy. METHODS: The EDLS concentrations in following samples: (1) villi in the 1st trimester (2) placenta, amnion and umbilical jelly of Wharton in the 3rd trimester, (3) plasma of normal non-pregnant women (4) maternal plasma in three trimesters, 24-48 hours post partum, (5) maternal plasma and umbilical blood in labour, were determined by radioimmunoassay. RESULTS: The concentration of EDLS in villi, placenta, amnion and jelly of Wharton was about 37, 33, 29 and 24 times higher respectively than the concentration of maternal blood. There was a significant positive correlation between the concentrations of EDLS in placenta or amnion and that in jelly of Wharton. Plasma EDLS levels in pregnant women of 1st and 2nd trimesters were similar to that in non-pregnant women, while in the 3rd trimester of pregnany they were significantly increased, and then significantly decreased in 24 to 48 hours post partum. There was a significant positive correlation between the plasma EDLS level and mean arterial pressure in the 3rd trimester. The EDLS concentration in plasma of umbilical vein and artery was about 3.1 and 2.9 times higher respectively than that in the maternal plasma. There were significant positive correlations between plasma EDLS levels of umbilical vein, artery or maternal vein and the weight of placenta or newborn. CONCLUSION: The fetal placenta unit is the source of EDLS which increases in maternal vein plasma during normal pregnancy. PMID- 10806731 TI - [Preliminary study on bone loss rate in early and late stages of post-menopausal women]. AB - OBJECTIVES: To investigate the characteristics of bone loss rate and bone turnover in early and late stages of normal chinese postmenopausal women, and to evaluate the predictive value of bone biochemical markers. METHODS: Thirty-two early postmenopausal women (1-3 years after menopause, group 1) and 33 late ones (15-30 years after menopause, group 2) were enrolled. Serum and bone alkaline phosphotase (alkaline phosphotase, ALP; bone alkaline phosphotase, BAP), fasting uriuary calcium/creatinine ratio (Ca/Cr), and Pyridinoline/creatinine ratio (Pyd/Cr), serum osteocalcin(BGP), bone mineral density (BMD) were measured at laseline and 6 months, 12 month follow-up. RESULTS: In group 1, mean lumber mineral density decreased 1.3% and 2.6% at 6 and 12 month follow-up as compared with initial values, In group 2, bone loss rates of femoral neck and Ward's triangle were 1.3%, 5.3% at 6 months, and 1.9%, 4.6% at 12 months respectively. Urinary Pyd/Cr elevated with time in the two groups. Changes of BGP, BAP in the two groups were similar. There were positive correlations between ALP, Ca/Cr, BGP, BAP and BMD. No correlation was found between bone loss rate of lumber spine and biochemical markers. CONCLUSION: Lumber spine was the favorable site to assess rapid bone loss in early postmenopausal women. Femoral neck and Ward's triangle region were superior to other site in predicting slow bone loss. Bone turnover of the two groups were all accelerated. Single measurement of biochemical marker failed to predict bone loss rate. PMID- 10806732 TI - [The effect of insulin-like growth factor-I on steroidogenesis and ultrastructures of cultured human granulosa cells in vitro]. AB - OBJECTIVE: To observe the effect of insulin-like growth factor-I (IGF-I) on steroidogenesis and ultrastructures of cultured human granulosa cells in vitro and the synergetic influence of follicle stimulating hormone (FSH) on this process. METHODS: Human granulosa cells were cultured in serum-free 1640 medium. After stimulated with various concentrations of IGF-I or IGF-I + FSH for 24 hours, estradiol (E2) and progesterone (P) levels in the media were measured by radioimmunoassay, and the ultrastructure changes of granulosa cells were observed under electronic microscope. RESULTS: When stimulated with different concentrations of IGF-I for 24 hours, E2 and P levels produced by granulosa cells were in a dose-dependent manner and significantly higher than those of controls (P < 0.01). The E2 and P levels in the media under the stimulation of IGF-I + FSH were 3.5 times and 14.5 times respectively higher than those of FSH alone. The main ultrastructureal appearances of granulosa cells stimulated with IGF-I were abundance of enlarged smooth enodplasmic reticulum, mitochondria and lipid droplets in cytoplasm. CONCLUSIONS: IGF-I possesses a positive effect on the steroidogenesis of cultured human granulosa cells in dose-dependent fashion which could be enhanced by combination of IGF-I and FSH. The ultrastructures of granulosa cells showed a tendency toward activation of steroidogenesis under the stimulation of IGF-I. PMID- 10806733 TI - [Failure of mifepristone induced interruption of pregnancy: point mutation at genetic codon 722 in human progesterone receptor gene]. AB - OBJECTIVE: To detect the point mutation of hormone binding domain (HBD) of human progesterone receptor (hPR) gene by molecular biological methods in women with insensitivity to mifepristone. METHODS: 26 women in 5 hospitals were consented in the study. 16 women were as study group: continuing pregnancy in 11 and missed abortion in 5. 10 women with successful abortion were as control group. Decidual tissue was collected in each woman and RNA was extracted from it and purified. Reverse transcription-polymerase chain reaction (RT-PCR), single strand conformation polymorphism(SSCP) and restriction enzyme reaction were performed. All women were followed up. RESULT: No point mutation of HBD of hPR gene was detected in control group except one with point mutation in trace. Point mutation of HBD of hPR gene was detected in 7 of women with continuing pregnancy. Mifepristone was administrated in midluteal phase in two of these women and no vaginal bleeding was observed. No mutation of hPR gene in the other women with continuing pregnancy. Three women had pregnancy again in 6 months and had successful abortion. The other one administrated mifepristion in midluteal phase and had second vaginal bleeding. No point mutation of hPR gene was detected in women with missed abortion. CONCLUSION: There is point mutation of HBD of hPR gene in women with insensitivity to mifepristone, and women with point mutation of hPR gene detected by molecular biological methods are really insensitive to mifepristone. PMID- 10806734 TI - [The clinical evaluation of endometrial carcinoma following irradiation for cervical cancer]. AB - OBJECTIVE: To study the characteristics and clinical features of endometrial carcinoma occurred after pelvic irradiation. METHODS: Clinical data of 27 cases of endometrial carcinoma occurred after irradiation for cervical cancer were reviewed, estrogen receptor (ER) and progesterone receptor (PR) status were studied in 17 cases by immunohistochemistry. RESULTS: The incidence of endometrial carcinoma occurred following pelvic irradiation was 0.19%. The mean age of patients was 63 year old. The mean incubation period from pelvic irradiation to diagnosis of the disease was 15.6 years. The proportion of advanced cases was 51.9% (14/27). The expression of ER and PR was very low (11.8%, 2/17). The original sites of 5 cases with stage I were in fundus or corner of the uterus, which received lower dose irradiation formerly. Operation was the best choice of treatment, 19 out of 27 cases received this modality. The survival rate of 3 and 5 year was 56.8%, 37.8%, respectively. CONCLUSION: The incidence of endometrial carcinoma occurred following pelvic irradiation for cervical cancer was higher than that of cancer occurred spontaneously, the proportion of advanced cases was high, the expression of ER and PR was very low, operation was the mainly method of treatment. The prognosis was poorer than that of spontaneous disease. PMID- 10806736 TI - [Advances in the research of the role of nitric oxide and nitric-oxide synthase system in the pathogenesis of pre-eclampsia]. PMID- 10806735 TI - [The clinical features and management of borderline ovarian tumors]. AB - OBJECTIVE: To study the clinical features and management of borderline ovarian tumor (BOT). METHODS: Retrospectively study 70 cases of BOT. RESULTS: Age 16-76 years old, average 45.6 year old; histological type: mucinous 46(65.7%), serous 21(30%); stage: I 46 (65.7%), II 1(1.4%), III 11(15.7%), IV 0%, relapse 12(17.1%). Tumor marker: elevated CA125 11/21, carcinoembryonic antigen (CEA) 6/10. TREATMENT: all patients were operated, 45 were given adjuvant therapy. OUTCOME: No evidence of disease (NED) 45, with evidence of disease (WED) 4, died of disease (DOD) 11, died of intercurrent disease (DID) 10. 11 patients who were given conservative surgery were NED, there were 2 long term CR among 16 patients with residual disease who were given adjuvant treatment. The 5, 10, 20 and 30 year survival is 90.0%, 87.1%, 87.1% and 85.6% respectively. COX hazard regression model analysis showed that histological type (r = 24.825), stage (r = 2.236) and residual disease (r = 2.083) were the main factors which impact on the prognosis of BOT. CONCLUSIONS: (1) The proportion of mucinous tumor is higher than that of serous tumor in this group; (2) Surgery is the proved only effective treatment of BOT till now, without any residual disease is expected; (3) Adjuvant therapy can't improve the prognosis of BOT; (4) CA125 and CEA can be two effective tumor markers in monitoring of BOT. PMID- 10806737 TI - [Current status of the study on maternal-infant transmission of hepatitis C virus]. PMID- 10806738 TI - [Urinary stress incontinence]. PMID- 10806739 TI - [Study of the liver changes by B-ultrasound and histopathology in severe pregnancy induced hypertension]. AB - OBJECTIVE: To investigate the liver changes in severe pregnancy induced hypertension (PIH) on B-ultrasound and pathological examination and to detect a possible connection with fatty liver of pregnancy. METHODS: From Jan 1992 to Nov 1997, 90 pregnant women in the third trimester were enrolled for study. They were divided into 3 groups: the pregnant women with liver diseases; mild and moderate PIH; and severe PIH (30 cases each). Ultrasonography of the liver was performed on each patient after admission. 15 percutaneous liver biopsies were performed under the guidance of real-time ultrasound scans on 15 severe PIH patients with or without liver dysfunction. Tissues were stained with specific lipid staining (Sudan) and with hematoxylin-eosin. All specimens were analyzed by light microscopy. RESULTS: Fatty liver was observed by B-ultrasound in 2 and 12 cases of the pregnancy with liver diseases group and the severe PIH group, respectively. 14 of the 15 liver biopsy specimens showed fatty infiltration of varying degrees. however, only 8 of the 14 cases were detected fatty liver by B ultrasound. CONCLUSIONS: These findings suggest that liver involvement in severe PIH is a pathological change. Severe PIH and acute fatty liver in pregnancy could have related pathogenesis components of the same. PMID- 10806740 TI - [Relationship between change of erythrocyte membrane lipid composition and pregnancy induced hypertension]. AB - OBJECTIVE: To study the cause of decreased erythrocyte deformability and its relation to pregnancy induced hypertension (PIH). METHODS: The cholesterol and phospholipid content of erythrocyte membrane in 24 non-pregnant healthy women, 22 normotensive pregnant women and 36 cases with PIH were determined using automatic biochemical analysor, Gas chromatography and modified colorimetric method. RESULTS: The cholesterol content and the cholesterol/phospholipid (C/P) ratio of the erythrocyte membrane were obviously higher in the group with moderate or severe PIH than that in the nonmotensive pregnancies (P < 0.01). There was no significant difference between mild PIH group and the normotensive pregnant group (P > 0.05). CONCLUSION: The increase of cholesterol content and C/P ratio in the erythrocyte membrane might be involved in the pathophysiology of PIH. The alteration of cholesterol content in the erythrocyte membrane probaby reflected the disturbance of serum lipid metabolism. PMID- 10806742 TI - [Different types of acidemia at birth, fetal heart rate patterns and infants outcome at four years of age]. AB - OBJECTIVE: To study the relations between different types of acidemia in umbilical artery at birth, fetal heart rate patter'ns (FHR) during laobr and infant long-term outcome. METHODS: 2,981 cases with singleton term pregnancy were selected for retrospective case-control study. Fetal monitoring was performed periodically during labor and umbilical blood gas was analyzed at birth and the infants outcome was followed up at age 4. RESULTS: There were 47 infants with pure metabolic acidemia and 67 infants with mixed acidemia. The incidence of FHR score < 6 and 6-7 at the end of first stage was significantly higher in the two study groups than that in the control group. Late deceleration was associated with pure metabolic acidemia and complicated variable deceleration with mixed acidemia. Both types of acidemia were correlated with Apgar scores below 7 at 1 minute after birth. Developmental screening at age 4 showed no significant differences between the two study groups and the control groups. In the mixed acidemia group there were slightly more infants had deficits in speech/language development. CONCLUSIONS: Late deceleration may be an indicator of pure metabolic acidemia and complicated variable deceleration be an indicator of mixed acidemia. Both types of acidemia were correlated with Apgar scores below 7 at 1 minute after birth. The relation between acidemia at birth and deficit in speech/language development should be further evaluated. PMID- 10806741 TI - [Uterine and placental vascular lesions in pregnancy induced hypertension and its relationship to pregnancy outcome]. AB - OBJECTIVE: To study the vascular lesions of the uterus and placenta in severe pregnancy induced hypertension (SPIH) and its relationship to pregnancy outcomes. METHODS: 29 cases of SPIH and 29 cases of normal term pregnancies (NTP) were examined to observe the expressions of IgA, IgG, IgM and C3 in the vascular walls of placental villus and uterus placental bed by immunohistochemical technique. RESULTS: (1) In SPIH, the positive expression rates of the IgA, IgG, IgM and C3 in the vascular wall of placental villus (20.1%, 24.1%, 72.4%, and 65.5%) and uterus placental bed (20.0%, 20.0%, 70.0% and 60.0%) were significantly higher than those of NTP (all P < 0.05), while there were no expression in NTP. (2) The thickening of endangium angiostenosis, fibronoid necrosis and acute arterial atherosis were found at the sites of positive expressions in SPIH. (3) In SPIH complicated by intrauterine growth retardation (IUGR), the positive expressions of the immunocomplex and complement in the vessel walls (76.2%) were significantly higher than those of the SPIH without IUGR (50%, all P < 0.05). CONCLUSION: In SPIH, there are depositions of immunocomplex and complement on the vessel walls of placental villus and uteroplacenta. Immunological factor plays an important role in the development of PIH. PMID- 10806743 TI - [The prognostic value of heart rate variety on congestive heart failure of pregnant women]. AB - OBJECTIVE: To explore the prognostic value of heart rate variety (HRV) on congestive heart failure (CHF) of pregnant women complicated with organic heart disease. METHOD: One hour ambulatary electrocardiography was performed in 61 pregnant women complicated with organic heart dieases at about the 28th-32th week in gestational period and 7 days after delivery respectively. RESULT: 9 patients occured CHF later, their HRV time domain index were significantly decreased (P < 0.001, P < 0.05), compared with patients without CHF as control group. 7 days after delivery, When the condition of CHF patients was improved. Their SD, SD SD value increased. CONCLUSION: HRV has some value of assessing the cardiac reserve and predicting the happen of Congestive Heart Failure in pregnant women complicated with organic heart disease. PMID- 10806744 TI - [The effect of 25 micrograms misoprostol on induction of labor in late pregnancy]. AB - OBJECTIVE: To determine the efficacy and safety of 25 micrograms misoprostol for labor induction in the third trimester of pregnancy. METHODS: 48 term pregnant women with single pregnant vertex present intact membrane were selected in study group. All case in study group indications for labor induction, without contra indication of labor induction and misoprostol. The 48 women were randomly assigned A(25 micrograms) and B(50 micrograms) misoprostol groups. Misoprostol was placed in the posterior vaginal fornix every 4 to 6 hours. The maximum dose in 24 hours is 200 micrograms. Misoprostol was not given after either spontaneous rupture of membrane or beginning of active labor. RESULT: The sucesess rate of induction labor was groups 77.8%, 81.0% in group A and B, respectively. The average duration from administration of misoprostol to start of labor in two groups was 796.9 +/- 359.9 minutes and 807.4 +/- 405.2 minutes, respectively; the average duration from start of misoprostol use to vaginal delivery was 978.6 +/- 464.4 minutes, 977.5 +/- 421.4 minutes, respectively. The incidence of vaginal delivery had no statistics significant difference between two groups (P > 0.05). The incidence of hypertonic uterine contraction or hypertonic contractionchysystole with abnormal pattern of fetal monitoring in group A was lower than that of group B, but there was no significant difference (P > 0.05). CONCLUSION: 25 micrograms misoprostol is an efficient and safe dosage for labor induction in term pregnancy. PMID- 10806745 TI - [Study on the karyotype and recurrence risk of malformed newborns]. AB - OBJECTIVE: To survey the karyotype and recurrence risk of malformed newborns. METHODS: Karyotypes of 104 malformed newborns were analyzed using G-banding. All cases were devided into two groups depending on their karyotypes: Group 1 with normal karyotype (n = 51) and group 2 with abnormal (n = 53). Clinical data were compared between malformed groups and control. Women who had the second babies after delivering a malformed (n = 20) were included in the statistics of recurrence risk. RESULTS: (1) Of 104 cases, 53 had abnormal karyotype (50.96%). They were trisomy 21, trisomy 18, 45X, 1q+ and 5p+ (n = 41, 9, 1, 1, 1, seperately). (2) Within group 2, women's mean delivery age was elder, while premature delivery rate was higher than that of control (P < 0.01, 0.05). In group 1, the proportion of women with abnormal pregnancy history increased as compared with control (P < 0.05). The rate of fetal distress, poly/oligohydramnios and small for gestational age (SGA) of two malformed groups was higher apparently than that of control. The birth weight of two malformed groups declined obviously. (3) Within 20 second babies being born after their malformed siblings, 6 were malformed too. The total recurrence rate was 30%. CONCLUSION: (1) Abnormal karyotype is an important cause of congenital malformations; (2) The age pregnant women abnormal pregnancy history, IUGR and poly/oligohydramnios all may be the high risk factors of congenital malformations; (3) The recurrence rate of malformations rises after the first malformed newborn's birth. PMID- 10806746 TI - [Influence of fetal distress on neonatal behavior neurological assessment and its prognosis]. AB - OBJECTIVE: To study the short and long term prognosis of fetal distress. METHODS: 60 newborns with fetal distress (study group) and 60 normal newborns (control group) were assessed by neonatal behavior neurological assessment (20 items) in neonatal period and by reformed wechsler intelligence scale children-revised at 2 years of age. RESULTS: The total scoring of study group was significantly different from that of control group (P < 0.01). In the study group, there were 41 cases without neonatal asphyxia whose scoring was significantly different from that in the normal group (P < 0.05), whereas the scoring of other 19 cases with neonatal asphyxia was significantly lower than that of the control group (P < 0.001). We also found that the scoring of 38 acute fetal distress cases was significantly different from that of 22 chronic cases (P < 0.01). In two years follow-up study, the scoring of children's intelligence test in acute distress cases was not different from that of controls, but the scoring of intelligence test in chronic distress cases was markedly different from that of controls (P < 0.01). CONCLUSION: Fetal distress could affect neonatal behavior and their intellectual development. PMID- 10806747 TI - [A study of maternal and fetal peroxidation in elderly primigravidea]. AB - OBJECTIVE: To analyze the rlationship between the age of pregnant women and maternal and fetal peroxidation. METHODS: Pregnant 164 women in their third trimester were collected and divided into 3 groups according to maternal ages: in group 1, maternal age less than 35, n = 71; group 2, maternal age 35-39, n = 63; group 3, 40 or over, n = 30. All the women had no other high risk factors. Maternal and umbilical blood samples were drawn during delivery, and the concentration of superoxide enzyme (SOD) and Lipid peroxidate produce (LPO) were determined. RESULTS: There were no significant differences in neonatal Apgar scores, birth weight, and the volume and quality of amniotic fluid among the 3 groups, but the cesarean scetion rate increased along with maternal age. The fetal SOD concentration was significantly higher than that in maternal serum, but the LPO level was lower. There was significant correlation between maternal age and maternal serum SOD and LPO levels (r1 = 0.15, P < 0.05; r2 = 0.2299, P < 0.01). The maternal LPO levels in group 2 and 3 were significantly higher than that in group1 (P < 0.05), and the concentration of SOD in group 3 was significantly lower than that in group 1 (P < 0.05). CONCLUSION: The activity and concentration of free radicals were higher in mother than that in fetus, and the ability of anti-oxidation was lower in mother. The maternal age is associated with SOD and LPO levels during pregnancy. PMID- 10806748 TI - [Antenatal diagnosis of albinism fetuses by fetalscopy]. AB - OBJECTIVE: To popularize the clinical utility of fetalscopy in antenatal diagnosis of albinism fetuses. METHODS: Fetalscopy examination was carried out in 85 high-risk pregnancy women, during 17 to 26 gestational weeks. Guided by the ultrasonography, the fetalscopy was inserted into the aminotic sac. Albinism fetuses was diagnosed by the color of the fetal hair. RESULTS: Fetalscopy were succeed in 79 of 85 high-risk women (93%). 63 fetuses were diagnosed as normal, 16 fetuses was diagnosed of albinism. There were 2 false-positive cases at 17 weeks of gestation due to mistaking wooly hair for hair. CONCLUSION: Fetalscopy is a quick, accurate, safe method in antenatal diagnosis of heredity diseases, such as albinism. PMID- 10806749 TI - [The study on the relationship between luteal phase serum sex hormone levels and pregnancy outcome in in vitro fertilization embryo transfer cycle]. AB - OBJECTIVE: To study the influence of maternal endocrine factors on clinical pregnancy outcome in an in Vitro Fertilization Embryo Transfer (IVF-ET) program. METHODS: 62 IVF-ET treatment cylces were studied. Luteal phase serum sex hormone levels [estrogen (E2), progesterone (P), prolactin (PRL)] in natural and superovulation cycles were measured by radioimmunoassay and the clinical pregnancy outcome was observed. RESULTS: Serum luteal phase sex hormone levels were higher than in natural cycles (P < 0.05). P level and P/E2 ratio in progesterone supplementation group were higher than the group without progesterone supplementation (P < 0.05). In pregnancy group serum E2(4,885.5 +/- 1,269.0 pmol/L) was lower, PRL (66.4 +/- 26.8 micrograms/L), P/E2 (373.0 +/- 152.5) were higher than in nonpregnancy group (E2 6,091.2 +/- 1,522.1 pmol/L, PRL 49.1 +/- 24.1 micrograms/L, P/E2 227.4 +/- 116.7) (P < 0.05). When P/E2 ratio was between 300-400 and PRL level was between 60-100 micrograms/L, the clinical pregnancy rate increased markedly (P < 0.05). CONCLUSION: Luteal phase serum sex hormone level may influence embryo implantation in IVF-ET. In order to improve the clinical pregnancy rate, it is very necessary to supplement progesterone for the IVF-ET patients according to the different sex hormone levels. PMID- 10806750 TI - [The expression of integrin beta 3 in cycling and early pregnant endometrium and its relationship with primary unexplained infertility]. AB - OBJECTIVES: To profile the changes in integrin beta 3 expression in cycling and early pregnant endometrium and its relationship with unexplained infertility. METHOD: Expression of integrin beta 3 on endometrium was detected in 56 normal cycling women during different menstrual phases or early pregnant women, and 9 cases with unexplained infertility during mid-luteal phase by immunohistochemistry technique. RESULTS: Integrin beta 3 was expressed only in the midluteal or pregnant endometrium(after cycle day 19). Compared with normal women, integrin beta 3 failed to express in the midluteal endometrium from women with unexplained infertility. CONCLUSIONS: Our findings suggest integrin beta 3 may play important roles in the process of implantation and decidualization. It might be a new reliable marker of uterine receptivity. Abnormal endometrial integrin beta 3 expression might be the major cause of unexplained infertility. PMID- 10806751 TI - [A clinical control study on the treatment of uterine leiomyoma with gonadotrophin releasing hormone agonist or mifepristone]. AB - OBJECTIVE: To compare the results and side effects in treating uterine leiomyoma with gonadotrophin releasing hormone agonist (GnRH-a) or mifepristone. METHODS: 75 patients with uterine leiomyoma who had clinical symptoms and diagnosed by Bcan were divided into two groups. The GnRH-a group (30 patients) was treated by injection of GnRH-a 150 micrograms/day subcutaneously for three months, and the mifepristone group (45 patients) was treated by mifepristone 12.5 mg/day po for three months. RESULTS: The clinical symptoms improved obviously in both groups. The volume of leiomyoma reduced 20.0% or more in 90.0% (27/30) of the patients in GnRH-a group, while it was 91.1% (41/45) in mifepristone group. However, the recurrent rates were 40.0% and 17.8% in the 2 groups. CONCLUSION: It suggested that mifepristone is a more practical and hopeful drug in treating uterine leiomyoma. PMID- 10806752 TI - [The changes of sex steroid receptors and morphology in Norplant treated endometrium]. AB - OBJECTIVE: To study the relationship between the changes of sex steroid receptors as well as endometrial morphology and abnormal uterine bleeding induced by Norplant. METHODS: Immunohistochemistry, Western Blot, in situ hybridization and computer image analysis were employed to demonstrate the expression of estrone receptor(ER) progesterone receptor(PR) protein and mRNA as well as morphometric analysis of the endometrium in Norplant users. RESULTS: The content of DNA and the numbers of spiral arterioles decreased in Norplant treated endometrium. The percentage of glandular area in Norplant endometrium was much lower than that in cyclic endometrium. A statistically significant correlation was observed between the percentage of area of glands and abnormal uterine bleeding, as well as serum estrogen(E2) level, but E2 level had no direct relationship with the bleeding abnormality. As the time growing, the percentage of glandular area increased. Immunostaining and relative concentration of glandular and stromal ER in Norplant treated endometrium were significantly weaker and less than all the stage of cyclic endometrium. However, PR, especially stromal PR immunostaining was very intense, which was the same or more than that in all the stage of cyclic endometrium. In situ hybridization detected that the ER and PR mRNA was lower in Norplant treated endometrium than that in cyclic endometrium. The ER mRNA expression was more less abundant than PR mRNA expression. CONCLUSION: These results suggested that poor repair and regeneration of endometrium(especially glands and spiral arterioles) and abnormal expression of ER and PR may be related to pathogenesis of abnormal uterine bleeding induced by Norplant implant. PMID- 10806753 TI - [Separation and isolation of fetal cells from maternal blood and its uses in non invasive prenatal diagnosis]. PMID- 10806754 TI - [Trends of the selection of hysterectomy routes]. PMID- 10806755 TI - [Clinical significance of pelvic lymph node dissection in the early stage of endometrial carcinoma]. PMID- 10806756 TI - [Molecular cloning of EXT2 and EXT4 gene]. AB - Hereditary multiple exostose(EXT) is an autosomal dominant disorder of skeletal system. Three genetic loci have been identified at 8q24.1(EXT1), 11p11(EXT2) and 19p(EXT3) respectively. In this paper, EXT2 gene was cloned with positional cloning and homologous screening. SSCP and sequencing analysis have been done in 37 EXT patients who came from 20 EXT families, 2 mutations of insertion were tested in 2 patients. This confirmed that the gene cloned in this paper was EXT2 gene which locus at 11p11. Additionally EXT4 gene was cloned with homologous screening and located at 1p36.1 with FISH in this paper. PMID- 10806757 TI - [Effects of retionoic acid and GM-CSF on apoptosis of HL-60 cell induced by mild heat]. AB - The effects of several factors on the apoptosis of HL-60 cells, a human acute myeloid leukemia cell line, were investigated. The apoptosis model was established with mild heat. Typical changes of cell morphology and characteristic DNA ladder pattern were found. The effect of apoptosis induced by mild heat was enhanced, when HL-60 cells were pretreated with the retinoic acid. By contraries, that of apoptosis was displayed with the slight inhibition while HL-60 cells were pretreated with rhGM-CSF. We also observed the alterations of mRNA expressions of c-myc by the dot hybridization. The results indicate that both the mild heat and the combination of mild heat and retinoic acid obviously reduce the mRNA expression of c-myc compared to the control group. PMID- 10806758 TI - [Modulation of endothelin-1 on pulmonary surfactant synthesis in lung explants]. AB - The effect of endothelin-1(ET-1) on pulmonary surfactant(PS) synthesis was studied in cultured lung explants without serum. The results showed that: 1. ET-1 increased [3H] choline incorporation with dose-dependence and time-dependence. 10(-10) mol.L-1 ET-1 increased the contents of total phospholipids, phosphatidylcholine and phosphatidylglcerol in lung tissues. The major membrane characteristic phospholipids, such as phosphatidylethanolamine, phosphatidylinostitol, phosphatidylserine and sphingomyelin, did not show changes. 2. BQ123, a ETA receptor selective antagonist, could decline the [3H] choline incorporation induced by ET-1(10(-12) and 10(-10) mol.L-1) (P < 0.01). 3. PMA, a protein kinase C(PKC) activator, increased [3H] choline incorporation into lung tissues (P < 0.01); the PKC inhibitor H7 decreased [3H] choline incorporation induced by ET-1(P < 0.01). The results demonstrated that ET-1 at physiological level can enhance the PS synthesis mediated by ETA receptor and PKC. PMID- 10806759 TI - [Chemosensitivity test for 170 human breast carcinoma samples]. AB - We tested the in vitro sensitivity of 170 primary breast carcinoma samples to the anti-cancer drugs conventionally used in clinical chemotherapy. It was found that: 1. The high sensitivity rate (HSR) of breast carcinoma cells to 5 fluorouracil (5-Fu), mitomycin (MMC), cisplatin (DDP), adriamycin(ADM), methotrexate(MTX), vincristin(VCR), vepesid(VP-16), taxol, and vindesine(VDS) were 57.1%, 49.4%, 48.3%, 35.8%, 21.8%, 21.2%, 10%, 18.2% and 20.9%, respectively. The increase of HSR of cancer cells to 5-Fu, MMC, DDP, taxol was more significant than that of the other drugs tested (P < 0.05). 2. No correlation was shown between drug sensitivity and factors, i.e. pathological type, clinical type, expression of estrogen receptor (ER) and epithelium growth factor receptor(EGFR) for breast carcinoma (P > 0.05). 3. Cyclosporin A can increase part of the sensitivity of ADM-resistant breast carcinoma cells to ADM. PMID- 10806760 TI - [Analysis of amino acids, vitamins and inorganic elements in Dictyophora indusiata]. AB - Fifteen amino acids(total content: 13.37 mg.100 ml-1), 12 inorganic elements(Zn 37.3 micrograms.g-1, Mn 56.9 micrograms.g-1, Cr 4.88 micrograms.g-1, Fe 370 micrograms.g-1, Se 0.81 microgram.g-1, Cu 28.2 micrograms.g-1, Co 0.78 microgram.g-1, Ni 2.09 micrograms.g-1, etc.), and vitamin E(73.2 micrograms.g-1) in Dictyophora indusiata were determined. Seven amino acids, including valine 0.61, leucine 1.04, isoleucine 0.66, threonine 0.74, methionine 0.20, lysine 0.52, and pheuylalanine 0.64(mg.100 mg-1), were found to be essential. The possible medical significance of Dictyophora indusiata is discussed. PMID- 10806761 TI - [Laboratory diagnosis of Chlamydia trachomatis and Ureaplasma urealyticum of chronic prostatitis]. AB - Prostatic fluid taken from 1038 cases of chronic prostatitis were detected by fluorescent monoclonal antibody technique and isolating cultural method for Chlamydia Trachomatis (CT) and Ureaplasma Urealyticum (UU). The control group consisted of 80 cases. CT and UU were not found in control group. The positive rates of CT and UU infection were 28.4% and 33.0% respectively in the tested group whose cases were abacterial prostatitis. The results suggested that CT and UU are pathogens causing chronic abacterial prostatitis. PMID- 10806762 TI - [Investigation of hepatitis B virus infection in blood donors]. AB - To investigate the status of HBV-M and HBV DNA in blood donors, 583 serum samples were detected by ELISA technique and polymerase chain reaction. The results were that hepatitis B virus marker(HBV-M) positive rate was 55.9%(326/583), HBV DNA 9.78%(57/583). The data indicate that there is high incidence of HBV infection in blood donors. The donors should be strictly tested for HBV in the future. PMID- 10806763 TI - [Distribution of erythrocyte EsD and PGM1 (phosphoglucomutase) phenotypes in the Han population of Changsha district]. AB - Phenotyping of erythrocyte EsD and PGM1 enzymes was performed by a mixed agarose starch gel electrophoresis in 165 healthy blood donors of Changsha District. The gene frequencies and cumulative discrepancy power(CDP) were calculated. The phenotype frequencies of EsD and PGM1 were compared not only with the different Chinese nationalities but also with those reported by the other countries showing a good correlation with the phenotype distributions of Han people in the other cities of China. The result showed that the gene frequencies of erythrocyte EsD1 and PGM1(1) were 0.6242 and 0.7121 in Han population of Changsha District, respectively. This method is now used routinely in our laboratory and is superior for the examination of individuality and identification of paternity. PMID- 10806764 TI - [Effects of nao yi-an granule on hemorheological indexes and RCD (erythrocyte deformability) in patients with hemorrhagic stroke]. AB - The effects of Nao Yi-an granule(Chinese medicine group, CMG) and western medicine (western medicine group, WMG) on hemorheological indexes and red cell deformability(RCD) were investigated in 40 patients with hemorrhagic stroke. The results showed that whole blood relative viscosity(high and low shear), whole blood reduced viscosity(high shear), plasma relative viscosity and RCD were significantly elevated before the treatment in both group than those of the control group(P < 0.01). After the treatment, the above indexes were lower in CMG than before the treatment and also lower in CMG than in WMG(P < 0.01). It is indicated that the Nao Yi-an granule is obviously superior to western medicine on the decrease of blood viscosity, increase of RCD, improvement of blood perfusion of brain tissue, protection of brain tissue and improved clinical in patients with hemorrhagic stroke. PMID- 10806765 TI - [Changes of hemodynamics, oxygen delivery and oxygen consumption during anesthesia induction with propofol in patients with rheumatic mitral stenosis]. AB - Changes of hemodynamics, oxygen delivery and consumption were studied in 20 patients with rheumatic mitral stenosis, undergoing mitral valve replacement, anesthesied induction with propofol(Group P) or midazolam(Group M). The results showed that hemodynamic values were decreased after intravenous propofol, and recovery after tracheal intubation. Hemodynamic values also decreased and last to 2 min after tracheal intubation in Group M. The reduction degree of HR in Group P were more, the degree of SVRI,. LVSWI, RVSWI less than those in Group M respectively. Oxygen consumption decreased more than delivery. It is suggested that induction of anesthesia with propofol in patients with rheumatic mitral stenosis is safe, and may be advantageous than midazolam. PMID- 10806766 TI - [A primary study of microanatomy on blood supplement of large and huge pituitary adenoma]. AB - OBJECTIVE: To explore the blood supplement regularity of large and huge pituitary adenomas. MECHODS: Sixteen large and huge pituitary adenomas were removed by microsurgery via transorbitofrontosphenoidal approach. The blood vessels of the tumors coming from surrounding structures were observed, photographed, videotaped and measured during operation. RESULTS: The blood supplement of the pituitary adenoma comes from not only the superior and inferior hypophyseal arteries, but also a lot of new arteries originating from meninges, optic nerves, optic chiasm, and arteries which surround the tumors, such as internal carotid artery, anterior cerebral artery and soon. CONCLUSION: The blood supplement of large and huge pituitary adenomas is increased obviously. The operation results will get much better if this situation is recognized and handled very well. PMID- 10806767 TI - [Relationship of lipoprotein(a) to fibrinolytic activities in healthy subjects]. AB - Plasma tissue-type plasminogen activator(tPA) and plasminogen activator inhibitor(PAI-1) and lipoprotein(a) [Lp(a)] in 133 healthy subjects were determined. The results demonstrated that PAI-1 activity was higher in Lp(a) > or = 0.3 g.L-1 group than Lp(a) < 0.3 g.L-1 group (P < 0.05). Univariate statistical analyses showed Lp(a) concentration was positively associated with PAI-1 activity (P < 0.01), but not with age, sex, body mass index, blood lipids or tPA activity. PMID- 10806768 TI - [Endoscopic variceal ligation combined with partial splenic embolization: preliminary clinical results]. AB - Endoscopic variceal ligation combined with partial splenic embolization (EVL-PSE) was performed in a group of 13 patients with esophageal variceal bleeding and hypersplenism due to portal hypertension from January 1997 to March 1998. PSE was performed one to two weeks before or one week after initial EVL, and a range of 30% to 60% of the splenic parenchyma was embolized. Repeated EVL was performed at two week intervals until the varices were eradicated. Active bleeding in the nine patients was successfully controlled and all the varices of the 13 patients were eradicated after EVL-PSE. Eradication of the varices required two to five(mean 3.1) EVL sessions, follow-up ranging from 2 to 16 months(mean 6.9 months). In all but one case, no rebleeding occurred. All patients after PSE showed a good response on peripheral blood cell count and reduction of splenomegaly. No major complication or death related to the combination therapy was observed. Preliminary results in this study show that this combination therapy may result in more rapid eradication of the varices and reduce rebleeding after endoscopic variceal ligation. However, more data and studies may be necessary for further evaluation. PMID- 10806769 TI - [Meckel's diverticulum and its complications in children]. AB - Seventy-two cases of Meckel's diverticulum were treated surgically in recent four years. There were 61 boys and 11 girls with a mean age of 5.2 years. Of the 65 symptomatic patients, 31(43%) had rectal bleeding, 16(23%) suffered from bowel obstruction and 17(24%) peritonitis. Patients with diverticulitis(8 cases) had acute abdominal pain in the periumbilical region or in the right lower quadrant compatible with appendicitis. Ectopic mucosa was found in 47 cases(65.3%). Gastric(36 cases) and pancreatic(11 cases) were the most common ectopic tissues. A nuclear medicine Meckel scan was positive in 26 of 31 patients(84%). It is emphasized that 99mmTcO4- scintiphoto is more specific in diagnosis of the Meckel's diverticulum. The best choice of the surgery is resection of the diverticulum and end-to-end intestinal anastomosis. PMID- 10806770 TI - [Clinical evaluation of cardiac troponin I in ischemic heart diseases]. AB - The detection of cardiac troponin I(cTnI), including serum or plasma, was evaluated in 114 patients with ischemic heart diseases or other heart diseases. The results were that the sensitivity of cTnI detection(qualitative analysis, cutoff is 0.2 ng.ml-1) was higher than that of creatine kinase(CK), creatine kinase-MB(CK-MB), alpha-hydroxybutyrate dehydrogenase(alpha-HBD), lactic dehydrogenase(LDH), and aspartic transaminase(AST) for diagnosing acute myocardial infarction (AMI) (93.2% vs 68.2%, 68.2%, 65.9%, 65.9%, and 75.0%; P < 0.05; respectively). The specificity of cTnI detection was higher than that of alpha-HBD, LDH, and AST(95.2% vs 81.0%, 73.8%, and 54.0%; P < 0.05; respectively) and similar to CK and CK-MB(95.2% vs 83.3% and 83.3%; P > 0.05; respectively). On the other hand, the sensitivity of cTnI in patients with unstable angina pectoris was 45.5%. It was higher than stable angina pectoris and lower than AMI. The results suggest that the cardiac troponin I is a better cardiac injury marker than other cardiac markers for diagnosing AMI. PMID- 10806771 TI - [Detection of interleukin-8 and nitric oxide in chronic prostatitis]. AB - We detected levels of interleukin-8(IL-8) and nitric oxide(NO) in 62 prostatic fluids. The results were that microbiologic exam, IL-8 content, and NO level in control group were all negative; there was not significant difference on the microbiologic exam and NO level between the chronic prostatitis group and non prostatitis group; there was significant difference between two groups of IL-8 positive ratio. These results suggest that the IL-8 detected in the prostatic fluid of patients with prostatitis may be a diagnostic indicator of prostatic infection. PMID- 10806772 TI - [Effect on endothelin and cytokines on the pathogenesis of coronary heart disease]. AB - The plasma levels of endothelin(ET), serum levels of tumor necrosis factor(TNF) and interleukin-6(IL-6) in 35 patients with coronary heart disease(CHD) and 20 healthy people were studied by radioimmunoassay and enzymebinded immunosorbent assay(ELISA). The results showed that the plasma ET level and the serum TNF level in the CHD group, especially the acute myocardial infarction(AMI) group, were much higher than those of the control group (P < 0.01). There was no obvious statistical difference on IL-6 levels between the narrowed coronary artery group and the control group (P > 0.05). But the serum level of IL-6 in the AMI group was significantly higher than that of the other groups (P < 0.01). These findings indicate that ET and cytokine levels may play an important role on the pathogenesis of coronary heart disease. PMID- 10806773 TI - [Effects of propofol and etomidate on cerebral oxygen supply and demand balance during middle hypothermia cardiopulmonary bypass]. AB - Effects of propofol(Group P) and etomidate(Group E) on cerebral oxygen supply and demand balance during middle hypothermia cardiopulmonary bypass(CPB) were studied in twenty patients undergoing cardiac valve replacement. Arterial blood and jugular bulb blood were taken to analyse blood gas, and cerebral oxygen content were caculated. RESULT: During hypothermia, jugular bulb oxygen saturation(SjO2), jugular bulb blood oxygen content(CjO2) increased; cerebral oxygen extraction rate(CEO2) decreased significantly in two groups(P < 0.05). During rewarming SjO2, CjO2 were significantly decreased, CEO2 increased compared with that before rewarming and hypothermia in two gbroups (P < 0.05). In Group P, SjO2 and CjO2 were higher, CEO2 lower than those in Group E respectively during rewarming (P < 0.05). CONCULUSION: The activity of propofol reduced cerebral metabolism, cerebral oxygen consumption and CEO2 is better than that of etomidate. PMID- 10806774 TI - [Cerebrospinal fluid cytology of acute viral encephalitis and meningitis]. AB - Using cytological examination, the cerebrospinal fluid(CSF) of 98 patients with acute viral encephalitis was investigated. We found 81.4% activated lymphocytes and 5.8% plasma cells in CSF in the first week. In addition, various lymphoid features were disclosed in association: mitosis, binucleation, abnormal nuclear lobulation, Russell bodies, and morula in plasma cells. These cellular changes specifically existed in CSF of acute viral encephalitis, which were helpful to distinguish the said disease from tuberculous melitis and multiple sclerosis. PMID- 10806775 TI - [Effects of serum bilirubin on lipoproteins]. AB - The effects of serum bilirubin on blood lipids and lipoproteins in 237 subjects were investigated. The findings were that total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C) were negatively correlated with total bilirubin and direct bilirubin. The results indicate that bilirubin affects metabolism of lipoproteins and the low level of serum bilirubin is a new risk factor for coronary heart disease. PMID- 10806776 TI - [Effects of orthodontic treatment on the shape of upper dental arch of patients after surgical operation of cleft lip and palate]. AB - Occlusal casts were measured and analysed in the following groups: orthodontic group(9 subjects treated orthodonticly after surgical operation of the cleft lip and palate), untreated group (10 subjects untreated orthodonticly after surgical operation), normal group(10 young subjects with normal occlusion). The results were that the upper dental arch length and width increased evidently (P < 0.01) in the orthodontic group and almost returned to normal; in contrast, the upper dental arch length and width were inhibited in the untreated group; there were significant differences among the normal occlusion group, the orthodontic group, and untreated group (P < 0.01). The results suggest that during the period of growth and development, the shape of the upper dental arch of the patients can be evidently improved by the orthodontic treatment after the surgical operation of cleft lip and palate. PMID- 10806777 TI - [Complications of endoscopic sinus surgery: incidence and prevention-analysis of 307 cases]. AB - The complications of endoscopic sinus surgery performed on 307 patients with chronic inflammatory paranasal sinus diseases were analysed. The results showed that: 1. The common complications were excessive bleeding, orbit penetration, intra-orbital haemorrhage, postoperative stenosis of the maxillary ostium and middle turbinate adhesions. 2. The most common perioperative complications were those related to orbital penetration except excessive bleeding. A major complication was a case of cerebrospinal rhinorrhea with secondary purulent meningitis. 3. The annual overall incidences and the rates of common complications have been significantly decreased with year. The causes and prevention of excessive bleeding, stenosis of maxillary ostium and middle turbinate adhesions are discussed emphatically. In addition, the proposals were offered concerning the diagnostic criteria of the complications, and the statistical method of the complication incidence. PMID- 10806778 TI - [VP-16 and VM-26 plus platinum drugs combined with radiotherapy in the treatment of small cell lung cancer]. AB - OBJECTIVE: To compare the clinical efficacy of VP-16(etoposide) with VM 26(teniposide) both plus platinum drugs combined with radiotherapy in the treatment of patients with small cell lung cancer(SCLC). METHODS: Seventy-four cases of SCLC were studied retrospectively. Forty-three patients were treated with VP-16 + DDP + CTX or VP-16 + DDP, the other 31 patients were treated with VM 26 + DDP or VM-26 + Carboplatin. The treatment of both groups was combined with radiotherapy. RESULTS: There was no significant difference between VP-16 and VM 26 groups in response rate, 2-year-survival rate, and brain metastasis rate. But the blood toxicity was more serious in VM-26 group than in VP-16 group; 3-4 stage toxicity rate was 58.06% versus 27.91%. CONCLUSION: The results indicate that both VP-16 and VM-26 combined with platinum drugs were effective chemotherapy protocols. The authors suggest choosing VP-16 protocols as the first choice for previously untreated SCLC patients. PMID- 10806779 TI - [Non-invasive detection of endothelial dependent dilatation dysfunction in patients with essential hypertension]. AB - With high-resolution ultrasound, we measured the diameter of the brachial arteries at rest, during reactive hyperaemia (with increase flow causing endothelium-dependent dilatation), and after sublingual glyceryl trinitrate(GTN; causing endothelium-independent dilatation) in 61 subjects, consisting of 30 controls with normal blood pressure(NP) and 31 patients with established essential hypertension(EH). The results showed that in patients with EH, flow mediated dilatation in arteries was much reduced for comparison with control group(P < 0.01). However, there was no significant difference in response to GTN between two groups(P > 0.05). It indicates that endothelium-dependent vasodilation is impaired in patients with essential hypertension. PMID- 10806780 TI - [Total cavo-pulmonary connection for complex congenital heart disease]. AB - Between April 1994 and January 1997, 10 patients with complex congenital heart disease were treated with total cavo-pulmonary connection(TCPC). They included 7 males and 3 females, aged 2-17 years(mean 10.1 years). These cases were single ventricle in 5, tricuspid atresia in 4, severe Ebstein anomaly in 1. For construction of the right atrial lateral tunnel, Gore-Tex patch was used in 9 cases and autologous right atrial wall in another, 4 of the 10 patients underwent TCPC without aortic crossclamping. There were no operative deaths. The follow-up is 2-36 months(mean 12.3 months). All of these patients did well and were in NYHA class I. The advantages, operative indication and technique of TCPC and the experience of improvement of surgical results are briefly discussed in this paper. PMID- 10806781 TI - [Determination and significance of nitric oxide and tumor necrosis factor-alpha induced by peripheral blood monocyte in vitro in patients with malignant bone tumors]. AB - Peripheral blood monocytes from 15 cases of malignant bone tumors and 12 cases of health volunteers were seperated and induced by lipopolysaccharide(LPS) for 48 hour culture. The supernatant was collected for determining the content of nitrite and tumor necrosis factor-alpha(TNF-alpha) (by Griess and ELISA methods, respectively). The results were as follows: The content of nitrite and TNF-alpha in patients with malignant bone tumors was significantly higher than that in normal control; the content of nitrite and TNF-alpha in the malignant bone tumor group presented the positive correlation (r = 0.8909, P < 0.01). The results suggest that NO and TNF-alpha produced by activated macrophages may be related to the pathogenesis of malignant bone tumors. PMID- 10806782 TI - [Influence of insufficient zinc on immune functions in NIDDM patients]. AB - The serum zinc level and immune functions were analyzed in 34 patients with NIDDM before and after the treatment with zinc gluconate supplement during conventional therapy (after the blood glucose level stabilization). The results showed that before treatment the level of serum zinc and red cell C3b receptor rosette(RBCK C3b RR), T-lymphocyte subgroup CD3, CD4, and CD4/CD8 were decreased(P < 0.01), while CD8, red cell immune complex rosette(RBC-ICR) were increased. After treatment with zinc gluconate for 1 month the serum zinc level, RBC-C3b RR, RBC ICR, CD3 and CD4/CD8 became normal, CD8 also approached to normal. All the above figures were significantly different before and after zinc therapy. The data showed that various degrees of lowering of serum zinc and abnormal immune functions were present during the conventional antidiabetic therapy. Thus, zinc supplement should be used as an important adjunctive therapy for NIDDM patients. PMID- 10806783 TI - [Expression and significance of tumor suppressor gene WAF1 in epithelial ovarian neoplasms]. AB - The expression of P21WAF1 protein in epithelial ovarian carcinoma, borderline ovarian tumor, benign ovarian tumor and normal ovary were analyzed by immuno histochemistry technique. Results indicated: 1. The positive expression for p21 was 45.7%(16/35) in epithelial carcinoma, 75%(9/12) in borderline tumor and benign tumor, 90.9%(10/11) in normal ovary. The difference for p21 protein between ovarian carcinoma and borderline tumor, benign tumor, normal ovary was obvious(P < 0.05). The differences in other groups were unimportant(P > 0.05). 2. The positive expression of p21 in stage I/II was 42.9%(6/14), in stage III/IV was 47.6%(10/21). The difference of them was insignificant(P > 0.05). 3. The positive expression for p21 in Grade I was 70%(7/10), in Grade II 50%(6/12), in Grade III 23.1%(3/13). The difference for p21 protein between Grade III and Grade I, Grade II was obvious(P < 0.05). The difference between Grade I and Grade II was unimportant(P > 0.05). 4. The positive expression for p21 in 35 ovarian carcinoma with lymphaden metastasis(17.6%) was lower than ovarian carcinoma without lymphaden metastasis(72.2%). The difference of them was obvious(P < 0.05). The conclusion is that deregulation of p21 protein is associate with tumoral differentiation, lymphaden metastasis and patient's prognosis, whereas it isn't associated with clinical stage. The loss for p21 protein may effect the onset and progress of ovarian carcinoma. PMID- 10806784 TI - [Determination of tryptophan in human serum by high-performance liquid chromatography]. AB - In this paper a high-performance liquid chromatographic(HPLC) method has been developed to determine the concentration of tryptophan(Trp) in human serum. Serum sample was precipitated with trichloroacetic acid solution(0.6 mol.L-1) to remove protein and centrifuged and assayed by HPLC. The operating conditions were NOVA PAK C18 column(3.9 mm x 150 mm), KH2PO4(0.1 mol.L-1) mobile phase with flow rate of 0.7 ml.min-1, and detective wave-length at 295 nm. The linear range was 1.223 2447.381 mumol.L-1. The detection limit was 0.612 mumol.L-1. The average recovery was 96.3%. Intra-assay CV is less than 4% and inter-assay less than 5%. The concentration of Trp in normal adult serum was 46.603 +/- 5.108 mumol.L-1. The detection time of serum Trp was within 5 min. The results suggest that this method is simple, fast, accurate, and convenient. PMID- 10806785 TI - [Behavior pattern and personality in patients with epilepsy]. PMID- 10806787 TI - [Retrospective analysis of 20 cases of occupied diseases in spleen]. PMID- 10806786 TI - [Value of intervention guided by common ultrasonic probe in the diagnosis of complicated abdominal masses]. PMID- 10806788 TI - [A broken sharp scalpel left in an intervertebral space and slipped to pelvic cavity during an operation of intervertebral disc displacement]. PMID- 10806789 TI - [A case of brain hernia induced by hemodialysis]. PMID- 10806790 TI - [A case of Crohn's disease complicated with Behcet's syndrome]. PMID- 10806792 TI - Augmentation of recombinant fibronectin polypeptide CH50 on the antitumor function of macrophages. AB - We prepared an anti-metastatic polypeptide, recombinant fibronectin polypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the function of macrophages. CH50 can significantly augment the production of nitric oxide(NO) by macrophages in a dose-dependent manner. The continuous presence of CH50 had a much stronger effect. In the presence of CH50, the cytotoxicity of macrophages to melanoma B16/F1 cells was significantly enhanced, and a stronger effect was obtained if CH50 was present continuously. CH50 polypeptide and IFN-gamma have a synergistic effect on the production of NO by macrophages and the cytotoxicity of macrophages on tumor cells. In the in vivo experiments, CH50 can inhibit the growth of tumor cells, and have a better effect in the presence of IFN-gamma. Our results suggest that recombinant fibronectin polypeptide CH50 has two functions: one is to inhibit the metastasis of tumor cells, and the other one is to augment the function of macrophages. And this polypeptide will be potentially useful in tumor therapy. PMID- 10806793 TI - Expression of vascular endothelial growth factor in hepatocellular carcinoma and its relationship to tumor growth and metastasis. AB - To understand the relationship between the expression of vascular endothelial growth factor (VEGF) and the growth, metastasis of hepatocellular carcinoma (HCC), immunohistochemistry and Northern blot were used to investigate VEGF protein and mRNA in 21 cases of HCC with and without metastasis. VEGF protein was found in 8 of 9 cases with metastasis, whereas only in 4 of 12 cases without metastasis. The positive rate of the former was significantly higher than that in the latter. VEGF mRNA was detectable in both carcinoma and its surrounding liver tissues, but its level in the former was 2-3 times higher than that in the latter. In carcinoma with metastasis, the mRNA level was 5-6 times higher than that without metastasis. It is concluded that VEGF is closely related to the growth of HCC as well as its metastasis and it might be a useful indicator for the metastatic potential of HCC. PMID- 10806791 TI - Expression of anti-CD4 human/murine chimeric antibody and their killer tumor activity. AB - From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were analyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse Ig heavy subgroup II (A) and kappa chain subgroup III, respectively. The VH and VL genes were subcloned into p gamma 1-Expr and p kappa-Expr respectively, then transfected into XL2-Blue. The VH- p gamma 1 and VL- p kappa were transfected by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques. We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies. These chimeric antibodies are able to kill tumor cells specifically in vitro. PMID- 10806794 TI - Differential effects of d, l-sotalol and d-sotalol on isoproterenol-increased delayed rectifier outward potassium current in guinea pig single ventricular myocytes. AB - The aim of this study was to compare the effects of d, l-sotalol and d-Sotalol on the delayed rectifier K+ outward current in the presence of isoproterenol at different concentrations. Time-dependent delayed rectifier K+ outward currents were measured in isolated guinea pig single myocytes using the whole-cell configuration of the patch-clamp technique. Currents were measured in response to 300 ms depolarizing pulses from a holding potential of -40 mV in three experimental protocols [control, isoproterenol (10(-9) mol/L-10(-6) mol/L), and isoproterenol (10(-9) mol/L-10(-6) mol/L) plus either d, l-Sotalol (10(-4) mol/L) or d-Sotalol (10(-4) mol/L)]. IK tail currents were measured upon repolarization to -40 mV. It was found that IK was significantly amplified in the presence of isoproterenol (10(-9) mol/L-10(-6) mol/L) plus d-Sotalol. At 10(-8) mol/L isoproterenol, IK was increased by 92.7% +/- 17.1% (P < 0.05) and 54.3% +/- 13.4% after d-Sotalol addition (P < 0.05). In contrast, d, l-Sotalol completely conteracted the increase of Ik by isoproterenol (< 10(-8) mol/L), and compared to control, IK was decreased by 35.6% +/- 8.1% at 10(-8) mol/L isoproterenol plus d, l-Sotalol (P < 0.05). It is concluded that the beta-adrenergic blocking property of d, l-Sotalol but not that of d-Sotalol maintains the delayed rectifier K+ outward current blockade in the presence of isoproterenol in guinea pig myocytes. This might contribute to a superior antiarrhythmic efficacy as compared to d Sotalol. PMID- 10806795 TI - Study on the hypercoagulable state in patients with angina and myocardial infarction. AB - The molecular markers of platelet activation, coagulation and fibrinolysis were detected in 60 cases of coronary heart disease (CHD), including 15 cases of stable angina (SA), 21 cases of unstable angina (UA) and 24 cases of acute myocardial infarction (AMI). The results showed that the platelet granule membrane protein 140 (GMP-140) level increased obviously in CHD groups compared with normal control, suggesting that platelet activation existed in CHD. Prothrombin fragment F1 + 2 and fibrinopeptide A (FPA) were examined to observe the activation of coagulation. No difference was found between SA group and normal controls, while their levels in both UA group and AMI group were significantly higher than in normal control and SA group (both P < 0.05). D-D dimer and alpha 2-plasma inhibitor (alpha 2-PI) were detected to observe fibrinolytic state. The results showed that no difference existed between SA group and normal controls, while both D-D dimer and alpha 2-PI in UA group and AMI group were significantly elevated than those in SA group and normal controls (P < 0.05). PMID- 10806796 TI - Clinical study on chronopharmacokinetics of digoxin in patients with congestive heart failure. AB - Fluorescence polarization immunoassay was used to study the chronopharmacokinetics of digoxin in 10 patients with congestive heart failure (CHF) who also served as self-controls. Our results showed that the serum digoxin concentration reached peak value 1 h after taking digoxin at 7:00 a.m., but the serum digoxin concentration reached the peak value 2 h after taking digoxin at 4:00 p.m. The average serum digoxin concentration area under curve was greater and the best maintainable time of serum concentration within 24 h after taking digoxin at 4 p.m. longer than those at 7:00 a.m. The heart rates were obviously lower and the cardiac function was significantly improved in 4:00 p.m. group. PMID- 10806797 TI - Effects of pHGF on hepatocyte DNA synthesis after partial hepatectomy in rats. AB - The effects of pHGF on the changes of hepatocyte proliferative cycle and liver regenerative capacity after partial hepatectomized rats were observed by flow cytometry (FCM). The results were as follows: 1) S phase fraction (SPF) in group of normal rats (group A) accounted for 9.89% and increased gradually within 6 h, following a peak at 12th h or 36th h after operation, but in the group of pHGF treated rats (group B) the peak appeared at 24th h after operation; 2) Proliferation index (PI) of group A was 19.6% before partial hepatectomy, increased to 34.91% within 6 h and reached a peaks at 12th or 36th h after operation, and in group B the peak appeared at 48th h after operation. There were significant differences between two groups in SPF and PI (P < 0.01). The weight of liver began to increase 12 h after operation, and almost reached the preoperative weight 5 days after operation. These findings suggest that pHGF can promote the DNA synthesis and segmentation of hepatocyte. PMID- 10806798 TI - Expression of the tumor metastatic suppressor gene in mouse melanoma model: inverse association to metastatic potential. AB - Data obtained in experimental cutaneous melanomas have suggested that the nm23 gene may function as a metastasis suppressor gene. The nm23 level in 8 human cutaneous melanoma cell lines and 2 murine melanoma cell lines were examined. Each melanoma cell line was transplanted subcutaneously into the flank of nude mice, and the metastatic behavior was evaluated by counting lung tumor foci and by determining host survival time. It was found that expression of nm23 mRNA in human melanomas is correlated closely with reduced metastatic behavior in experimental animals and may serve as a sensitive prognostic indicator of malignancy and survival in patients with melanomas. PMID- 10806799 TI - An experimental study on the disorders of hepatic hemodynamics and changes of plasma histamine in dogs with fulminant hepatic failure. AB - The model of fulminant hepatic failure induced by acetaminophen was established in dogs to observe the changes of hepatic hemodynamics and plasma histamine levels in portal vein (PV), hepatic vein (HV), abdominal aorta (AA) and inferior vena cava (ICV). The results showed that the portal vein resistance (PVR) was elevated and portal venous blood flow (PVF) was decreased; hepatic artery resistance (HAR) was decreased and the hepatic artery blood flow (HAF), portal venous pressure (PVP), wedged hepatic venous pressure (WHVP) and inferior vena cava pressure (ICVP) had no changes. The histamine of the PV, HV, ICV and AA were all elevated after formation of fulminant hepatic failure. And the increasing wave of the HV was the highest. The increased histamine in HV may be mediated by H1 receptor causing the contraction of hepatic venulae, resulting liver sinusoid congestion, increasing PVR and decreasing PVF which exacerbate the liver cell damage. Moreover, the more severe liver damage, the more histamine was released, and a vicious circle may ensue. Our results also suggest the possibility of using H1 receptor antagonist to treat the disturbance of liver hemodynamics in severe acute liver damage. The increased histamine in systematic circulation as a vasodilator may lower blood pressure and accelerate heart beats. The increase of plasma histamine may play an important role in the changes of hepatic and systemic hemodynamics in fulminant hepatic failure. PMID- 10806800 TI - I-2190A is a potent immunosuppressive drug for vascularized heart transplantation in rats. AB - The effect of a new immunosuppressant-I-2190A was tested in a rodent heart allograft model. Grafts were transplanted to recipients heterotopically. There were 5 groups: group 1 received no immunosuppressive agents; group 2 was given CsA (2.0 mg/kg, i.p.); group 3 was administered I-2190A (0.1 mg/kg, i.p.) in carboxymethyl cellulose (CMC); group 4 received injection of I-2190A (0.5 mg/kg, i.p. in CMC); group 5 received the combination treatment of I-2190A (0.1 mg/kg) and CsA (2.0 mg/kg). Immunosuppressants were discontinued 14 days after operation. No statistically significant difference in grafts median survival time (MST) was found between group 2 (9.5 days) and group 1 (9 days). The MSTs of grafts in group 3 (22 days, P < 0.05), group 4 (> 100 days, P < 0.01) and group 5 (> 100 days, P < 0.01) were significantly prolonged compared with control group 1 (9 days). Our results suggest that I-2190A is a potent immunosuppressant able to significantly prolong heart allograft survival in rats after a short time treatment, Low-dose I-2190A could potentiate the effect of sub-therapeutic dose of CsA as well. PMID- 10806801 TI - Evaluation of nuclide renal dynamic image in diagnosing diabetic nephropathy. AB - Twenty non-insulin dependent diabetics were given nuclide renal dynamic image examination to investigate its significance in the diagnosis of diabetic nephropathy (DNP). The results showed that 1) the glomerular filtration was hyperfunctioned in the diabetics with less than 5 years course, whereas it was impaired in the diabetics with over 5 years course and 2) nuclide renal dynamic image could reveal the glomerular hyperfiltration without beta 2 microglobulinuria or microalbuminuria, suggesting that it is superior to the clinical biochemical parameters in the diagnosis, especially in the early identification of DNP. PMID- 10806802 TI - The protein expression of Bcl-2, Bax, Fas/Apo-1 in acute myeloid leukemia. AB - The protein expression of bcl-2, bax, Fas/Apo-1 in 19 cases of acute myeloid leukemia (AML) were investigated by Western blot and ApAAp techniques. High expression of bcl-2 protein was found in most of the AML cases, and some of the cases expressed Fas/Apo-1 and bax. High expression of bcl-2 protein was associated with a bad clinical prognosis and a poor response after intensive chemotherapy. Bax seemed to coexpress with bcl-2 and so appeared to be a bad prognostic factor instead of a good one. The expression of Fas/Apo-1 was inversely correlated with bcl-2 and seemed to be a good prognostic factor which may reflect the relative homeostasis of apoptotic pathway. It is concluded that apoptosis-induced pathways in AML often exhibit disturbance features. Coregulation of bcl-2, Bax and Fas/Apo-1 genes formed the apoptosis-induced pathway, which is the biological factor affecting response to chemotherapy. PMID- 10806803 TI - Gene transfer into hematopoietic cells of mouse and its in vivo expression after transplantation. AB - We have shown previously that high-efficient gene transfer can be attained in primary hematopoietic cells using liposome-mediated gene transfer strategy. In order to examine the stability of gene expression mediated by this gene transduction protocol, we observed the expression of marker gene in vivo by using bone marrow transplantation (BMT) to engraft lethally irradiated mouse with the genetically modified hematopoietic cells. The results showed that the mouse transplanted with appropriated number of transduced cells remained alive and healthy. The PCR analysis and G418 selection of the spleen colonies and bone marrow cells isolated from lethally irradiated animals 15 days and 30 days after injection of genetically modified bone marrow cells showed that the progeny cells of the transduced hematopoietic stem cells still contained and expressed the transduced genes, suggesting that the hematopoietic system is at least partially re-constructed by the stem cells with marker gene and that the stable expression of foreign genes in vivo can be attained by using this easy and harmless transduction protocol. These findings provide experimental basis for clinician to further investigate the biology of marrow reconstruction and the mechanism of leukemia relapse after BMT. PMID- 10806804 TI - Detection of minimal leukemic cells in cerebral spinal fluid of children with acute lymphoblastic leukemia using the polymerase chain reaction technique. AB - V delta 2D delta 3 rearrangements of T cell receptor (TCR) gene from cerebral spinal fluid (CSF) cells was detected for diagnosis and monitoring of central nervous system leukemia (CNSL) in children with acute lymphoblastic leukemia (ALL) using polymerase chain reaction (PCR) technique. 20 patients were studied and in 12 of them the results of PCR and dot hybridization with clonospecific probes were positive, showing the presence of minimal blast cells in CSF. Our study suggested that the PCR method is an effective tool for clinical diagnosis of CNSL and is much more sensitive than routine CSF examination. PMID- 10806805 TI - Effects of biliary tract pressure on dynamics of the Oddi's sphincter and its nerve mechanism. AB - In order to explore the effect of biliary tract pressure on Oddi's sphincter and the mechanism of development of high pressure of biliary tract during acute obstructive and suppurative cholangitis (AOSC), house rabbits were used to establish model of high biliary pressure in acute biliary duct caecus. It was observed that when the pressure of the acute biliary tract was increased to 8 kPa, the electric activity of Oddi's sphincter was obviously enhanced, the pressure of Oddioes sphincter increased remarkably (P < 0.05), and even constant spasm appeared with accompanying increase of discharge frequency of the right greater splanchnic nerves (P < 0.05) and progressive decrease of mean arterial pressure. However, when lidocaine of 0.6% was used to block the right celiac plexus, no above-mentioned reaction happened when the biliary tract pressure was increased again. The results indicated that the acute biliary tract obstruction might induce the contraction or spasm of Oddi's sphincter and bring about a vicious cycle. Its mechanism is related to splanchnic nerves reflection and it is one of important factors in the development of AOSC course. PMID- 10806806 TI - Clinical study of multi-drug resistance gene (MDR1) expression in primary ovarian cancer. AB - This study was designed to measure the multi-drug resistance gene (MDR1) mRNA content and analyze clinical relationship between MDR1 expression and drug resistance in primary ovarian cancer. Reverse transcription PCR (RT-PCR) was used to measure MDR1 mRNA content in biopsy sample of 31 primary ovarian cancers (experimental group) and 30 gynecological tumors (control group). The level of 95.2% (20/21) MDR1 expression was relatively low, and the detected rate of MDR1 expression was 67.7% (21/31) in experimental group, which was higher than that in control group (40.0%, P < 0.05). The differences of MDR1 expression between the effective group and no effect group after combined chemotherapy was significant (P < 0.05). No significant relationship was found between MDR1 expression and clinical stage or histological classification or grade of differentiation in experimental group. We are led to concluded that primary ovarian cancers have drug-resistance clones which might express MDR1 spontaneously and expression of MDR1 may be used as a prognostic and predictive indicator for clinical response of ovarian cancers to combined chemotherapy. PMID- 10806807 TI - The diagnosis of diffuse goitre by ultrasound imaging in children. AB - In order to find an easy and accurate procedure for diagnosis of diffuse goitre in children, we examined 50 patients with diffuse goitre using fine needle aspiration biopsy cytology, thyroid antibody detection, thyroid hormone analysis and ultrasound imaging. In the meantime, 109 healthy children (control) were examined by ultrasound imaging. The results showed that thyroid imaging in health children was a smooth echo pattern with stronger homogenous echogram than surrounding muscle tissues. The patients with diffuse goitre showed an abnormally enlarged thyroid volume. In 22 (84.6%) of 26 children with chronic lymphocytic thyroiditis, a varied patch hypoechogenicity was found, of whom 18 (81.8%) had positive results of antibody testing. On the contrary, echo-pattern was normal in 17 (70.8%) of 24 patients with diffuse thyroid proliferation and only the remainder (7/24, 29.1%) had abnormal echo-pattern as well as elevated auto antibody titers, of whom 2 were confirmed as chronic lymphocytic thyroiditis by a repeat fine needle aspiration biopsy 1 year later. By using combined ultrasound imaging and antibody determination, 92% of the cases with chronic lymphocytic thyroiditis could be diagnosed. Our study suggests that ultrasonic imaging is an easy, non-invasive, reproducible and effective procedure in the differential diagnosis of chronic lymphocytic thyroiditis in children. PMID- 10806808 TI - Effect of fragment Asp1961-Glu1978 in fibronectin on the expression of triple domain polypeptide in E. coli. AB - Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690-Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 in E. coli was very low, but that of CH63 was very high. The results suggests that Asp1961-Glu1978 in FN is a key sequence influencing the expression of triple domain polypeptide in E. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography. Both of the cell-binding domains in the recombinant polypeptide were functional. The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function. PMID- 10806809 TI - The effect of neferine on foam cell formation by anti-low density lipoprotein oxidation. AB - Oxidatively modified low density lipoprtein (LDL) plays an important role in atheroslerosis (AS) development. To investigate the role of neferine (Nef) in anti-LDL oxidation and foam cell formation, the lipoprotein was derived and subjected to three different treatments: N-LDL (normal LDL), Cu2+ + LDL and Cu2+ + Nef + LDL. The LDLs were put at 25 degrees C for 24 h and the thiobarbituric acid reactive substance (TBARS) values were determined. They were 0.57 +/- 0.2, 6.01 +/- 0.22 and 2.26 +/- 0.13 nmol/mg protein, respectively. The difference was very significant (P < 0.01) for each two groups by t test. Mouse peritoneal macrophage (M phi) were exposed to 50 micrograms protein/ml of Cu2+ + LDL and Cu2+ + Nef + LDL at 37 degrees C for 60 h. The tryglyceride (TG) and total cholesterol (TC) content in M phi were assayed. The results showed that Cu2+ + LDL was more efficient than Cu2+ + Nef + LDL in stimulating lipid accumulation in M phi (P < 0.001). The study demonstrated that Nef could inhibit Cu(2+)-mediated LDL oxidation and thereby inhibiting macrophage-derived foam cell formation. PMID- 10806810 TI - Voltage- and use-dependent effect of 7-chlor-benzyltetrahydropalmatine on sodium currents in guinea pig ventricular myocytes. AB - The whole-cell patch-clamp technique was employed to obtain information about the voltage-dependence and kinetics of interaction of 7-chlor benzyltetrahydropalmatine (7-Cl-BTHP) with cardiac sodium channels. 7-Cl-BTHP (30 mol/L) significantly decreased the peak sodium current (from 7.8 +/- 1.8 nA to 5.3 +/- 1.4 nA, P < 0.01, n = 5), without producing a shift of the current voltage curve. It shifted the inactivation curves of sodium current to hyperpolarized potentials, and the V0.5 was shifted from -(82.5 +/- 2.5) mV to (95 +/- 2.4) mV (P < 0.05, n = 4). 7-Cl-BTHP produced a significant use-dependent effect that was proportional to the duration of the voltage step. In addition, 7 Cl-BTHP slowed the recovery of sodium channel from inactivation, which could explain its use-dependent effects on sodium current. The characteristics of 7-Cl BTHP blockage suggest that this agent binds preferentially to inactivated sodium channels. PMID- 10806811 TI - The role of ras gene mutation in gastric cancer and precancerous lesions. AB - Abnormality of ras gene family was studied in a total of 206 cases of gastric cancer and precancerous lesions by PCR-RFLP, PCR-SSCP and DNA sequencing. The results showed that mutation rate of H-ras 12 codon in metaplasia, atypical hyperplasia, early-stage cancer and advanced cancer was 16.7%, 31.2%, 50.0%, and 32.2%, respectively. In the groups of superficial gastritis and normal controls, no mutation were detected in codon 12 of ras. Mutations of H-ras 61 codon and N ras 12 codon in various groups were the same as those in normal control. K-ras 12 codon mutation was detected in only 2 cases of gastric cancer by using PCR-SSCP, but it was not detected by DNA sequencing, which may be polymorphism. All H-ras 12 codon mutations were G-->T mutation. There were significant difference between the groups of metaplasia, dysplasia, gastric carcinoma and normal control group (P < 0.05, P < 0.01, P < 0.01, respectively). It was concluded that H-ras 12 codon mutation was an early event and may play an important role in gastric carcinogenesis. Although K-ras, N-ras mutation rates are high in colon cancer and leukemia, it seems to bear no relationship with gastric cancer. PMID- 10806812 TI - HSP60, HSP70 in the pathogenesis of Kawasaki disease: implication and action. AB - HSP60, HSP70 in plasma of 11 cases of Kawasaki diseases (KD) and 23 healthy children were determined. The two groups were controlled for age. Determination of HSP60, HSP70 was conducted in lymphocytes of 14 cases of KD and 26 healthy children. The results were compared with those of 12 patients with febrile diseases and 10 patients with tuberculosis. Our results showed that except a significant difference in plasma HSP70 found between acute phase and convalescent phase of KD (P < 0.01), no significant difference was found in HSP60, HSP70 among all groups (P > 0.05). The differences in HSP60, HSP70 in lymphocytes were relatively obvious among all groups. The levels of HSP60, HSP70 in acute phase of KD were significantly higher than those in convalescent phase or in healthy controls (P < 0.01). The levels of HSP60 in KD were significantly higher than those of patients with febrile diseases. HSP60 of KD children was significantly lower than those of children with tuberculosis (P < 0.01). The findings showed that HSP60, HSP70 might contribute to the pathogenesis of KD. Determination of HSP60, HSP70 in lymphocytes is of help in the diagnosis of KD. PMID- 10806813 TI - Extrahepatic and intrahepatic replication and expression of hepatitis C virus. AB - In order to determine the replication sites of hepatitis C virus, the in situ hybridization and immunohistochemical technique using digoxin-labeled 531 bp plus strand and minus-strand HCVRNA probes were employed to detect HCVRNA in the liver tissues, bone marrow mononuclear cells and peripheral blood mononuclear cells (PBMCs) from the patients with chronic hepatitis C, and in HCV transfected COS cells. The results showed that both plus-strand and minus-strand HCVRNA were detected in 80% of liver tissues (4/5). Plus-strand HCVRNA could be detected in 90% of PBMCs and bone marrow mononuclear cells (18/20), minus-strand HCVRNA in 25% of PBMCs. In HCV transfected COS cells, plus-strand HCVRNA distributed evenly in 20% cellular nuclei and cytoplasms. No minus-strand HCVRNA was detected in the bone marrow mononuclear cells and HCV transfected COS cells. The positive signal appeared in more cells when the liver tissues, PBMCs and marrow mononuclear cells were hybridized by plus-strand probes than when hybridized by minus-strand probes. Our results suggested that the hepatocytic cytoplasms and PBMC cytoplasms were the replication sites of HCV, but the marrow mononuclear cells were not the replication sites of HCV although they were infected by HCV. HCV infection might be accounted for the pathogenesis of chronic hepatitis and relapse of hepatitis C after liver transplantation. PMID- 10806814 TI - Cytomegalovirus infection in children's liver tissues. AB - Immunoperoxidase histochemical assay with monoclonal antibody against human cytomaglovirus (HCMV) was used to detect immediate early antigen (IEA) and early antigen (EA) of HCMV infection in liver tissue of 72 pediatric cases (34 autopsies and 38 biopsies). The HCMV antigen was positive in 25% (18/27). Among them, 12 cases were both HCMV-IEA and EA positive; 4 were HCMV-IEA positive and 2 HCMV-EA positive only. Liver HCMV infection rate in neonates, the infants with the age < 1 year and > 1 year was 8.0%, 60.0%, and 14.8%, respectively, indicating that liver HCMV infection occurred at various ages. The liver HCMV infection rate in different diseases was 50.0% in infantile hepatitis syndrome; 70.0% in extrabiliary malformation, and 12.5% in other hepatopathies, suggesting that infantile hepatitis syndrome and extrabiliary malformation were related with HCMV infection in liver tissues. PMID- 10806815 TI - Detection and identification of enteroviruses RNA by using polymerase chain reaction. AB - For rapid diagnosis of enteroviral infection in clinic practice, we developed a reverse transcription and polymerase chain reaction (RT-PCR) assay. Primers homologous to the conserved 5' non-coding region were designed by analyzing enteroviral genomes, and then they were used to enzymatically amplify RNA from 31 prototype enteroviral strains and enteroviruses (EV) in cerebrospinal fluid (CSF) of 34 cases of aseptic meningitis and 11 cases of aseptic encephalitis. The RT PCR products generated with these enteroviral primers were analyzed by agar gel electrophoresis and dot blot hybridization analysis. 31 EV strains showed an obvious monoclonal amplification band, and all dot blot hybridization results were positive. Four other viruses and cells cultured were all negative. The study of sensitivity of the RT-PCR showed that amplification production were positive to 10(-2)-10(-3) 50% tissue culture infective doses. With this assay, 21 (61.8%) of 34 aseptic meningitis and 8 (72.7%) of 11 aseptic encephalitis contained EV RNA in CSF samples. Two cases of meningitis and one of encephalitis with EV infection were still positive during convalescence. Our results suggest that this RT-PCR method was a fast, sensitive and specific technique for detection of common EV infection. PMID- 10806816 TI - Hormone secretion by cell culture of human GH-PRL secreting pituitary adenomas: effects of bromocriptine. AB - Dopamine agonists effectively reduce the secretion of prolactin (PRL) in the great majority of prolactinomas and reduce the bulk of the adenomas, as well as have partial therapeutic effect on some patients with acromegaly. The inhibitory effect of bromocriptine (BC), a dopamine agonist, on growth hormone (GH) and PRL secretion of dispersed cells from the pituitary adenomas of 16 cases of acromegaly, which secret GH and PRL simultaneously, were evaluated in vitro. The significant inhibitory effects of BC on PRL secretion were found in 12 cases. It was also found that PRL secretion was strongly inhibited when GH was suppressed; on the contrary, when GH secretion was not suppressed, the production of PRL was not or weakly inhibited. The exact mechanism of the effects is unclear so far. It is necessary to investigate, at molecular level, the etiology of GH-PRL adenomas and its response to therapeutic agents. PMID- 10806817 TI - The relationship between constitutive nitric oxide synthase (cNOS) and pregnancy induced hypertension. AB - To study the relationship between constitutive nitric oxide synthase (cNOS) and pregnancy induced hypertension (PIH), cNOS expression and localization in placental villi of PIH patients (n = 15) and normal pregnancy patients (n = 15) were immunohistochemically studied. The positive immunostaining of cNOS was located in trophocytes and cytoplasm of vascular endothelial cells. The positive rate in PIH patients was much higher than that in cases of normal pregnancy. The positive rates were 34.40% in mild, 44.74% in middle and 50.14% in severe PIH patients respectively. There was a significant difference in positive rate among the mild, middle and severe PIH patients (P < 0.01). It is concluded that the increase of cNOS activity probably was the results of protective or compensatory mechanism of PIH course. PMID- 10806818 TI - The role of vague nerve in the hemodynamic changes induced by acute biliary tract pressure increase. AB - In order to explore the mechanism of hemodynamic changes caused by high biliary tract pressure, we established an animal model of high biliary tract pressure, in which the disturbance of hemodynamics developed. The cervical or abdominal vague nerve was then blocked. It was observed that when the biliary tract pressure was increased to 16 kPa and kept for 1 h, the arterial blood pressure and cardiac output decreased immediately and parallelly (P < 0.05). When the cervical or abdominal vague nerve were blocked or the pressure of the biliary tract was decreased to zero, both indices returned to normal immediately (P > 0.05). The change of cardiac output lags a little behind that of arterial blood pressure. It suggests that the signal of biliary tract pressure increase can be sent to the cardiovascular center through vague nerve, and the balance between sympathic and parasympathic nerve was broken, which led to the weakening of cardiac contraction and decrease of cardiac output. Due to the peripheral effects of vague nerve, hemodynamic resistance of vessels decreased, which brought about redistribution of peripheral blood flow. Both were the causes of hemodynamic disturbances. After the blood pressure decreased markedly, it showed a jump to normal state when cervical vague nerve was blocked. And the amplitude of diastolic blood pressure restored more than that of systolic blood pressure. This suggests that the cardiac output and peripheral blood resistance are important factors that cause the decrease of blood pressure. PMID- 10806819 TI - The bacterial inhibitory ability and in vivo drug release pattern of a new drug delivery system: ciprofloxacine/tricalcium phosphate delivery capsule. AB - The bacterial inhibitory ability of a new drug delivery system (DDS): Ciprofloxacine/tricalcium phosphate delivery capsule (CTDC), its in vivo drug release pattern, and the influence of ultrasonic irradiation on its drug release were investigated. It was found that CTDC had a strong and sustained inhibitory ability to some common pathogens of bone and joint infections, such as staphylococcus aureus, escherichia coli and pseudomonas aeruginosa. In vivo drug release study in animals demonstrated a high concentration of ciprofloxacine in the bone tissue surrounding CTDC which was placed in the greater trochanter of the rabbit and continued to release ciprofloxacine for at least 5 weeks and the blood level of ciprofloxacine was low. In vivo study also showed ultrasonic irradiation could increase the amount of ciprofloxacine released from CTDC, which may be an economical, effecient and safe new method to achieve the control of drug release from DDS. PMID- 10806820 TI - Left to right extracardial shunt to control hemorrhage of ascending aorta and left ventricle: a report of 3 cases. AB - Presented in this paper are 3 cases of hemorrhage of ascending aorta and left ventricle after open heart surgery treated by extracardial bypass in our hospital from Oct. 1994 to Dec. 1995. Remained aneurysmal wall enclosing conduit graft was used as a sac bypassed to right atrium to form a extracardial left-to-right shunt in order to control bleeding and the results turned out to be satisfactory. The bypass and hemodynamically ignorable shunt can close spontaneously without complications with recovery of coagulation system. The technique may find wide application in clinical practice. PMID- 10806821 TI - Animal experimental study on surgical repair of hypospadias by free peritoneal graft. AB - Surgical repair of hypospadias was successfully performed by using free peritoneal graft in the model of rabbit hypospadias. The results showed that free peritoneal graft used as a substitute for urathra had a high survival rate, and the canal was formed well. Our study demonstrated that peritoneum could be used for the surgical repair of hypospadias and other urethral disorders such as urethral stricture. PMID- 10806822 TI - Cervical ripening in the third trimester of pregnancy with intravaginal misoprostol: a double-blind, randomized, placebo-controlled study. AB - To evaluate the safety and efficacy of intravaginal misoprostol for cervical ripening in the third trimester, a randomized, double-blind, placebo-controlled trial was conducted in 85 patients indicated for induction of labor and with unfavorable cervices. They were randomly assigned to receive either intravaginal misoprostol (100 mg) or placebo placed in the posterior vaginal fornix. The Bishop score, fetal heart rate and Doppler blood flow velocity waveforms were measured before and 12 h after drug administration. Placenta and decidu were histopathologically observed in some cases. Among 85 patients enrolled, 43 received misoprostol and 42 received placebo. Whereas the mean initial Bishop scores were not significantly different between the two groups, the mean Bishop score in misoprostol group was significantly better than those in placebo group. The mean change in Bishop score was also significantly different (4.4 for misoprostol versus 1.0 for placebo, P < 0.01). The prevalence of spontaneous onset of labor within 12 h after drug insertion in misoprostol group (67.4%, 29/43) was significantly higher than that in placebo group (14.3%, 6/42), P < 0.01. The average Doppler velocity systolic to diastolic (S/D) ratios of umbilical artery, middle cranial artery, renal artery were not significantly different before and 12 h after drug insertion between both groups. There was no significant difference in frequency of abnormal fetal heart rate tracings or fetal distress and in the mean Apgar scores between the two groups. Except the presence of vasodilation in villi vessels in the misoprostol group, the placental and decidual histopathological changes had no significantly difference between two groups. It is concluded that intravaginal misoprostol may be an effective and safe cervical ripening agent in the third trimester of pregnancy. PMID- 10806823 TI - An experimental study on the mechanism of anisodamini hydrobromidum in treating acute ischemic renal failure. AB - In order to explore the mechanism of anisodamini hydrobromidum (654-2) in treating acute ischemic renal failure, the model of acute ischemic renal failure in white New Zealand rabbits was established to dynamically observe and statistically analyze the intracellular concentration changes of free calcium ([Ca2+]i) and inositol triphosphate (IP3). The results showed that the levels of [Ca2+]i and IP3 in acute renal failure group were higher than those in control group (P < 0.01). However, the levels of [Ca2+]i and IP3 in 654-2 treated group were significantly lower than those in acute renal failure group (P < 0.001). It was concluded that 654-2 could alleviate Ca(2+)-overload in renal histocytes in acute ischemic renal failure. The protective mechanism is associated with intracellular reduction of IP3. PMID- 10806824 TI - Neglect and constructive disorder. AB - A 61 year-old right handed man, who suffered from right cerebral infarction with evidences of visual-spatial neglect and constructive disorder, was reported. When copying simple geometric designs, he omitted to copy figures on the left side of the page; he tended to bisect the line to the right of the line's real center; after memorizing the familiar pictures he mainly mentioned the pictures on the right side of the page; when copying the "Rey Complex" he also ignored the structures on the left side. The relations of the neglect and construction disorder are discussed. PMID- 10806825 TI - An observation on the mimic ecology of the oncomelania hupensis in three-Gorge reservoir areas. AB - According to the modulation of three-gorge-reservoir water level, the two observation spots were established at Gaoyang Town and Jianshe Village in Xingshan County of Hubei Province in order to observe the survival and reproduction of the oncomelania under different ecological conditions. Our one year observation revealed that most settled oncomelania died between June and August in the year and only a few of them survived the winter. The oncomelania hupensis's survival rate in group I, II, III settled in Gaoyang Town was 10%, 1%, 0%, respectively, and the oncomelania Sichuan's 12.5%, 22.5%, 13.5%. The rates obtained from spots in Jianshe Village is 0%, 2%, 0% and 9.5%, 19%, 3%, respectively. The survival rate of oncomelania Sichuan was higher than that of the oncomelania hupensis in the two spots. After draining the feeding pools, the soil samples were harvested and used for egg-hatching. The numbers of eggs that hatched in group I, II in Gaoyang were 1 and 2, respectively. The young oncomelania were found in each spot pool after the drainage, suggesting that the local condition is favorable to the oncomelania and the mating season of each group oncomelania is from April to August with the most active mating time being from May to June. PMID- 10806826 TI - Extraction of Phytolacca acinosa and its mulloscacidal effects. AB - The total saponins extracted from the plant root of Phytolacca Esculenta Van Houtte were studied for its mulloscacidal effect. The results showed the death rate of oncomelania was 95.6% under the concentration of 125 mg/L of saponins, at 28 degrees C with action time being 24 h. PMID- 10806827 TI - Expression of functional domain of chicken gizzard calponin. AB - A full-length cDNA of the function domain of wild-type chicken gizzard calponin was cloned into expression vector pAED4 and the recombinant function domain of wild-type calponin was expressed in an Escherichia coli expression system. The actin domain of calponin (CaP-B) can bind with actin and it is a requisite for its inhibition of ATPase and vasoconstriction of smooth muscle. In this study, the cDNA of CaP-B was inserted into vector pAED4 by direction-cloning method. The cDNA of CaP-B was obtained with PCR cloning technique. The recombinant DNA pAED4 Cap-B was transformed into E. coli BL21 (DE3) and identified with the restriction analysis. The bacterial clones containing transformants were induced to be highly expressed in E. coli BL21 (DE3). The target protein was detected and identified by Western Blot analysis. The content of target protein was as high as 10% of the whole protein after overnight (16 h) culture. The results confirmed that Cap-B was relatively highly expressed in E. coli. PMID- 10806828 TI - Effects of Huangdan capsule on plasma cGMP and ANP in rats with chronic renal failure. AB - The model of chronic renal failure (CRF) was made in 5/6 nephrectomized rats and Huangdan capsule was used to treat these rats. The levels of cyclic guanosine monophosphate (cGMP) and atrial natriuretic peptide (ANP) in plasma were examined. The results showed that Huangdan capsule could postpone the increase in the levels of cGMP and ANP, suggesting that by regulating the water and sodium metabolism, Huangdan capsule could ameliorate the glomerular filtration rate, and that Huangdan capsule could lower the levels of cGMP and ANP in plasma via body regulation. PMID- 10806829 TI - Effects of perindopril on vascular wall and left ventricular hypertrophy in rats with experimental hypertension. AB - The model of hypertensive rat was established by using the method of abdominal aortic constriction, 33 male SD rats were randomly divided into 4 groups: group A (sham-operated rats serving as controls), group B (hypertensive rats serving as controls), group C (hypertensive rats subjected to low-dose perindopril treatment, 0.1 mg.kg-1/d) and group D (hypertensive rats receiving high-dose perindopril treatment, 1.5 mg.kg-1/d). The drug was administered by gavage for 4 weeks. At the end of the treatment, mean arterious pressure (MAP) was measured via catheter in the carotid artery. The heart was taken and weighed to calculate the ratio of left ventricular weight to body weight (LVW/BW). The ratio of medial mean thickness to mean diameter of lumen of aorta and superior mesenteric artery was calculated by morphometry. Concentration of angiotensin II (A II) in plasma and in aortic wall were determined by radioimmunoassay (PA II, AA II). Our results showed that both high-dose and low-dose perindopril may prevent hypertrophy of left ventricle and hypertrophy of the tunica media of superior mesenteric artery. In addition, high-dose perindopril may significantly inhibit hypertrophy of aortic media. The above effects of perindopril may be explained by inhibition on tissue renin-angiotensin system of both vascular wall and heart, but is not related to circulating A II (PA II). PMID- 10806830 TI - [Regulation of myocardium beta-adrenoceptors pathway in ventricular remodeling of heart failure patients]. AB - To study the role of myocardium beta-adrenoceptors pathway in ventricular remodeling of heart failure patients. beta-adrenegic receptor density (Bmax) and the content of cAMP were measured in the papillae of left ventricle and blood lymphocyte of 20 patients suffered from heart failure (CHF) (NYHZ classification II to III) Bmax were investigated using 3H-dihydroalpheolol as ligand. cAMP were assessed by competitive immunoassay. Left ventricle mass index (LVMI) were measured using echocardiogram. The results showed that the Bmax and cAMP in failing myocardium significantly negatively correlated with LVMI (r = -0.77, P < 0.01 and r = -0.46 P < 0.05 respectively); the Bmax of myocardium and blood lymphocyte in CHF patients with NYHA III (63 +/- 12 fmol/mgpro and 514 +/- 115 fmol/10(7) cell) significantly lowered than that of NYHA II patients (94 +/- 20 fmol/mgpro and 702 +/- 138 fmol/10(7) cell); and the Bmax of myocardium and blood lymphocyte in patients with abnormal LVMI (62 +/- 12 fmol/mgpro and 516 +/- 122 fmol/10(7) cell) decreased more significantly than that with normal LVMI patients; even in nromal LVMI patients (92 +/- 21 fmol/mgpro and 682 +/- 146 fmol/10(7) cell), the Bmax of blood lymphocyte was already decreased (P < 0.01), when comparing with controls. The intralymphocyte cAMP content sygnificantly decreased than that of controls (P < 0.05). These results indicated that Bmax could reflect the severity of ventricle remodeling and the impairment of myocardium. The regulation of myocardium intracellular messenger transduction was earlier than the pathologic structural change of LV remodeling. PMID- 10806831 TI - Effect of low HDL combined with hypertriglyceridemia in coronary artery disease patients on PGI2 biological activity in relation to lipid regulating treatment. AB - The purpose of this study was to investigate the effect of low high-density lipoprotein (HDL) combined with hypertriglyceridemia in coronary artery disease (CAD) patients on prostaglandin I2 (PGI2) biological activity in relation to lipid regulating treatment. The inhibitory rate of PGI2 on ADP-induced platelet aggregation was used as the index for PGI2 biological activity. Twenty health individuals served as normal controls. CAD group consisted of 20 patients with low HDL combined with hypertriglyceridemia. The results showed that, before the treatment, the stabilizing effect on PGI2 activity decreased significantly in CAD group when compared with the control group (P < 0.01). One month after the treatment, HDL level in CAD group increased significantly and TG level significantly decreased (P < 0.001). The different effect on PGI2 activity was no longer found between CAD group and control group (P > 0.05), further confirming the protecting effect of HDL on PGI2 biological activity. Low HDL is considered as an important risk factor of CAD, therefore, impaired PGI2 biological activity and increased platelet aggregation might be responsible mechanisms. Furthermore, raising HDL level by lipid regulating treatment could restore the protective effect of HDL on PGI2 and might be helpful in the prevention of the acute coronary syndrome. PMID- 10806832 TI - Experimental studies of electroacupuncture on ventricular fibrillation threshold in rats with acute ischemic myocardium. AB - By ligating the proximate left anterior descend (LAD) of coronary artery and inducing the ventricular fibrillation with electrical stimulation, the preventive effects of electroacupuncture (EA) on ventricular fibrillation were observed. The results showed that the ventricular fibrillation threshold (VFT) of rats with acute ischemic myocardium was raised after acupuncturing some acupoints, which could prevent the occurrence of ventricular fibrillation. Furthermore, the combination of EA and propranolol could enhance the VFT effectively, and they showed a good synergistic effect. PMID- 10806833 TI - The regulating effects of protein kinase C on the tone of guinea-pig trachea and human lobus bronchi. AB - The regulating effects of protein kinase C (PKC) on the tone of guinea-pig trachea and human lobus bronchi were investigated by measuring the tone of isolated tracheal and bronchial strips. The effects of PKC on the tone of guinea pig tracheal and human lobus bronchi were observed and compared. The results showed that: (1) PKC activator PMA induced concentration-dependent relaxation in guinea-pig isolated trachea strips. This relaxation was completely ablated by the pretreatment with 5 x 10(-6) mol/L Ro31-8220 which is a PKC-specific inhibitor, but was not affected by the removal of epithelium (EP), or by the pretreatment with propranolol (beta-receptor blocker) or atropin (M-receptor blocker); (2) PMA led to concentration-dependent contraction in human lobus bronchi. This contractile response was completely depressed by the pretreatment with 5 x 10(-6) mol/L Ro31-8220 and was partly inhibited by 1 x 10(-5) mol/L isoptin (Ca(2+) antagonist), but was not significantly affected by propranolol or atropin. It is concluded that PKC is involved in the regulation of airway smooth muscle tone. The regulating effects may vary in different animals. PMID- 10806834 TI - Effect of aprotinin on the red cell immunity in cardiopulmonary bypass. AB - To investigate the effects of aprotinin on red cell (RC) immune function in the patients undergoing cardiopulmonary bypass (CPB), 20 patients who received valve replacement procedure were prospectively studied. The patients were randomly assigned to aprotinin group and control group. Red blood cell C3b receptor ratio (RC3bRR), red blood cell immune-component ratio (RICR), plasma C3, C4, CH50 and IgG level were determined before operation, at the end of CPB and 1st, 3rd, 7th postoperative days. Our results showed that: (1) The blood requirement was reduced in aprotinin group. (2) After bypass, plasma C3, C4, CH50 was maintained in aprotinin group, while they declined in control group. (3) Plasma level IgG fell in both groups, but at 7th postoperative day it recovered in aprotinin group while the level stayed at low level in control group. (4) The RC3bRR and RICR was mildly inhibited in aprotinin group. It is concluded that the impairment of RC immune function caused by CPB can be mitigated by aprotinin, which may be related to the effects of aprotinin in blood sparing, restriction of complement activation, and reduction of blood requirement which could mitigate the non specific inflammatory reaction. PMID- 10806835 TI - Study on the relationship between the Bcl-2/Bax ratio and the growth types of leukemic cells and drug resistance in acute myelogenous leukemia. AB - The ratio of Bcl-2 to Bax (Bcl-2/Bax) in 40 patients with acute myelogenous leukemia (AML) were determined. At the same time, the CFU-L of AML patients and drug resistance were detected by cell culture and MTT assay. The results showed that Bcl-2/Bax in AML was significantly higher than that in normal control (P < 0.001). Bcl-2/Bax ratio in colony growth group was higher than that in no-growth group (P < 0.05). Between drug resistance group and drug sensitivity group there was a significant difference in Bcl-2/Bax ratio (P < 0.05). There were more cases of drug resistance in high ratio group (H) than in low ratio group (L) (P < 0.05). Meanwhile, Complete Remission(CR) rate in group H was obviously lower than that in group L (P < 0.05) and patients in group H tended to have poor response. The above results indicated that the alteration of Bcl-2 and Bax is an important mechanism in the pathogenesis, progress and the development of drug resistance in AML. The determination of Bcl-2/Bax ratio has far-reaching implication in the treatment, choice of chemotherapeutic agents, and prediction of prognosis. PMID- 10806836 TI - Experimental study on the treatment of diabetes by phloridzin in rats. AB - Male rats at six weeks of age were divided into 5 groups at random: in group I, the rats with diabetes received 70% pancreatectomy; group II had sham-operation serving as controls; diabetic rats in group III were treated with phloridzin; In group IV rats received sham-operation and phloridzin treatment and group V were phloridzin-treated diabetic rats to be studied after discontinuance of phloridzin. 70 days after surgery, the weights and insulin contents of operated remnant pancreas were markedly higher than the expected value of 30%, reaching 44% (48.2% +/- 15.2%), demonstrating that the remnant pancreas still had capacities of compensatory regeneration and proliferation capacities. Phloridzin treated diabetic rats completely returned to normal in terms of oral glucose tolerance and insulin sensitivity. Discontinuation of phloridzin treatment in diabetic rats resulted in the recurrence of insulin resistance. These results suggested that normalization of hyperglycemia could ameliorate insulin resistance under diabetic conditions. PMID- 10806837 TI - [Action of DDPH in the interventional treatment of portal hypertension induced by liver cirrhosis in rabbits]. AB - To explore a new way of utilizing intervential treatment to effectively decrease the portal hypertension, the animal models of liver cirrhosis accompanying portal hypertension were set up by intraportal vein injection of suspensions of Bletilla striata to 10 rabbits by means of prospective investigational method. The catheter filled with heparin solution was remained in theportal vein for infusion of drugs. Three weeks later, DDPH solution was injected into the portal vein via the catheter to determine its effect of decreasing portal vein pressure and its influence of peripheral blood pressure and heart rate. At the same time, other 10 rabbits with liver cirrhosis associated with portal hypertension were subjected to the injection of DDPH solution via the ear vein served as control group. The results showed that administration of DDPH by portal vein could rapidly, safely and effectively decrease the portal hypertenive pressure without side effects and partially reverse liver cirrhosis, as compared with injection of DDPH via peripheral veins. It was concluded that DDPH exerted its alpha 1-adrenoceptor blocking and calcium antagonistic effects, thereby significantly reduce the portal hypertension. Injection of DDPH via portal vein is a completely new and relaible method for the treatment of liver cirrhosis with portal hypertension. PMID- 10806838 TI - Establishment of an animal model of reversible chronic partial obstructive hydronephrosis. AB - The establishment of an animal model of reversible chronic partial obstruction hydronephrosis was reported. The continuous ultrasonographic and pathological investigation demonstrated that this model fulfilled all the requirements, especially the reversibility. The technique is a relatively simple and reliable method. PMID- 10806839 TI - Glioma prostaglandin levels correlate with brain edema. AB - The present study was designed to prospectively investigated the prostaglandin (PG) levels and extent of peritumoral edema in 30 cases of glioma by using methods of radioimmunoassay and imaging. Both TXB2 and 6-keto-PGF1 alpha levels in all glioma groups went up over that in the control group. TXB2 level and ratio of TXB2/6-keto-PGF1 alpha were markedly increased with the extent of tumor malignancy. Water concentration in anaplastic astrocytoma and glioblastoma were significantly elevated. Difference in TXB2 level and TXB2/6-keto-PGF1 alpha ratio among three edema grades were statistically significant. TXB2 level and ratio of TXB2/6-keto-PGF1 alpha were closely correlated with water concentration (r1 = 0.53, r2 = 0.72, P < 0.01). Our findings suggested that the metabolism of PG in glioma were in the state of disorder, and that the imbalance between PGI2 and TXA2 may be one of factors which affect the formation of peritumoral edema. PMID- 10806840 TI - The inhibitory effects of IFN alpha-2b on EGFR expression and growth of cultured subconjunctival fibroblasts. AB - To investigate to what extent the growth of cultured subconjunctival fibroblasts was regulated by epidermal growth factor receptor (EGFR), we observed the expression of EGFR inhibited by interferon alpha-2b (IFN alpha-2b) and determined the growth of cultured subconjunctival fibroblasts by immunohistochemical and MTT methods. The linear relationships between the concentration of IFN alpha-2b and EGFR expression or growth were y = 1252.7-554.61 lgx (r = -0.98, P < 0.01), y = 0.523-0.0421 lgx (r = -0.93, P < 0.05) respectively. The concentration of IFN alpha-2b at doubled IC50 to EGFR (6.55 x 10(3) IU/ml) only inhibited the cell growth by 25.13%. The results suggested that the growth of subconjunctival fibroblasts may be controlled by many other growth factors because it was only partially inhibited by decreasing EGFR expression. PMID- 10806841 TI - The role for endothelin in mesangial proliferative glomerulonephritis. AB - To understand the effect of endothelin (ET) on the mesangium and matrix proliferation on the basis of hyperlipidemia, the model of mesangial proliferative glomerulonephritis (MSPGN) was established by intravenously giving calf serum albumin (250 mg/L) to the rabbits fed on feed containing 1% cholesterol for 4-8 weeks. Our results showed that in the 4th week of the experiment, the ET level in experiment group was significantly higher than that of normal controls (P < 0.01) and the pathological examination revealed development of MSPGN. In the 8th week of the experiment, the ET level was further elevated and pathological study demonstrated local glomerular sclerosis and infiltration of lymphocytes of interstitia. We are led to conclude that ET, as a newly identified hormone, is involved in the pathogenesis and development of MSPGN and acts as an important regulator in the proliferation of mesangium and accumulation of matrix of basal membrane. PMID- 10806842 TI - Clinical implication of serum sIL-2R levels in ovarian cancer. AB - The levels of serum soluble interleukin-2 receptor (sIL-2R) of 36 patients with ovarian cancer, 34 with benign gynecological tumors and 31 blood donors were measured by ELISA. Compared to those of benign and normal controls, serum levels of sIL-2R in patients with ovarian cancer elevated significantly (P < 0.01). Serum sIL-2R levels were likely related to types, clinical stage or histological grading of ovarian cancer. We were led to conclude that serum sIL-2R level may reflect the status of the immune system and the disease severity in patients with ovarian cancer. Its value as a marker for monitoring treatment and prognosis needs further studies. PMID- 10806844 TI - The effect of sheng bai solution on irradiated mice bone marrow cell division index and DNA content. AB - Before and after general irradiation with 60Co-gamma, mice were orally given Sheng Bai Solution (SBS) for one week. SBS alleviated the irradiation-induced reduction of bone marrow cell chromosome division index. The irradiation-induced decrease of marrow DNA amount, thymic and splenic fractions, and total leukocyte number were restored to some extent. SBS also helped to ameliorate general condition of patients. PMID- 10806843 TI - Effect of aluminum on long-term potentiation and its relation to L-arg-NO-pathway in hippocampal CA3 area of rats. AB - Experiments were performed on 64 Sprague-Dawley rats under urethane anesthesia. Extracellular recording method was used to investigate the effect of aluminum(Al) microinjected into CA3 on long-term potentiation(LTP) in this area. The relationship between the inhibitory effect of Al and L-arginine-NO pathway was also studied. Microinjection of Al (0.5 mol/L, 1 microliter) into CA3 could block the induction of LTP in CA3. Microinjection of Al (0.5 mol/L, 1 microliter) into CA3 after LTP was induced could also decrease the amplitude of population spike (PS). The inhibitory effect of Al on LTP in CA3 could be enhanced by preinjection of NG-nitro-L-arginine (0.3 mol/L, 1 microliter). Preinjection of L-arginine (0.3 mol/L, 1 microliter) into CA3 could antagonize the inhibitory effect of Al on LTP. These results suggest that Al could block the induction of LTP and decrease the amplitude of PS potentiated in CA3. The effect of Al might be antagonized by L-arginine-NO pathway. PMID- 10806845 TI - Fractal behavior of length change in human mitotic chromosome. AB - Sixty-six cells were examined for chromosome length change, with half of them (33) in the early-metaphase and the other half in the pre-metaphase. The 22 couples of chromosomes (excluding sex chromosomes) in each cell were analyzed. The differences of chromosome length between the two phases were demonstrated and the correlation dimension of each chromosome was calculated. It was suggested that the mitosis of human chromosome possessed the feature of chaotic decrease. This offers a new approach to the research of the dynamic process of the mitosis. PMID- 10806846 TI - Study on amino acid composition of HSP70 and the level of plasma free amino acids of workers with long-term exposure to harmful factors. AB - In order to know the practical value of heat stress protein 70 (HSP70) and to know the changes of plasma free amino acids of workers with the induction of HSP70 by harmful factors, the amino acid composition of major HSP, HSP70 purified from the heated cultured human leukemia cancer cell line K562, rabbit liver, rat liver and heart, and mouse liver with two-step procedures of DE52-cellulose ion exchange chromatography and affinity chromatography on ATP-agarose was examined. The level of plasma free amino acids of workers with long-term exposure to heat, carbon monoxide and the combined effect of both heat and carbon monoxide was also investigated. The results showed that the three richest amino acids in HSP70 of all origins were Gly, Glu and Asp, except that of rat heart which was rich in Gly, Phe and Glu. Additionally, Lys, Val, Leu and Ala were also found very rich in HSP70 of all origins. Compared with controls, the most of plasma free amino acids tended to increase and free methionine and tryptophan were increased significantly (P < 0.05) after a long-term exposure to heat, carbon monoxide, and both. These findings suggested that further studies need to be done to find the substances or drugs which induce the synthesis of HSP70 and reduce the inhibition of synthesis of normal proteins for the purpose of protecting people with exposure to harmful factors against the damage of the factors. PMID- 10806847 TI - Expression of intercellular adhesion molecule-1 in immune response of inner ear. AB - To understand the role of intercellular adhesion molecule-1 (ICAM-1) in immune response of the inner ear, inner ear immune response was induced in rats by inoculation of keyhole limpet hemocyanine (KLH) into the scala tympani of the animals who had been systemically sensitized. The expression of ICAM-1 in the inner ear was immunohistochemically examined. ICAM-1 was found in the epithelium of the spiral modiolar vein (SMV) with its collecting venules (CVs) as early as 6 h after challenge. Expression of ICAM-1 was observed on the epithelium of the endolymphatic sac (ES) and perisaccular region at 12 h. The intensity of ICAM-1 staining reached its peak within 24-48 h in these sites of the inner ear. By day 28, most specimens were devoid of appreciable staining for ICAM-1. Our study demonstrates that adhesion molecules play an important role in extravasation of inflammatory cells from the systemic circulation in the process of inner ear immune response. It also shows that cytokines that control expression of adhesion molecules may be released by cells outside ES, besides those cells in the ES. PMID- 10806848 TI - Embryonic limb buds derived neurotrophins on the survival of neurons and the growth of axons in culture in vitro. AB - Bioactive proteins from SD rat limb buds were extracted and purified. Fractions of 22 ku, 34 ku and 95 ku were proved to have neurotrophic activity to neurons, and the combined activity of these three fractions was the highest. So they were combinedly added into the culture medium of sensor neurons in dorsal root ganglia and motor neurons of anterior spinal cord from 2-week-old embryonic rats, and PBS was added as control. Phase-contrast microscopic and electron microscopic observations, and true cholinesterase measurements were performed to evaluate the survival and changes in growth, function, and ultrastructure of these cultured neurons. In the experimental group, it was found that the AchE activity was higher (P < 0.01), ultrastructural changes in mitochondria, Gorgi's complex and other cell organs were milder than those in the control group. The results showed limb buds derived neurotrophins played an important role in maintaining the survival of the neurons and promoting the growth of axons. It was concluded that embryonic limb buds derived neurotrophins had high neurotrophic activities on neurons' survival and axon growth. PMID- 10806849 TI - The expression of PDGF-B chain mRNA in lung tissue from rats repeatedly infected with mycoplasma pneumoniae. AB - In order to investigate the role played by platelet derived growth factor-BB (PDGF-BB) in the pathogenesis of pulmonary interstitial fibrosis in rats repeatedly infected with mycoplasma pneumoniae (MP), a rat MP infection model was developed by infecting rats with MP for 9 times during a period of 24 weeks with a technique of ultrasonic nebulizing inhalation. Then in situ hybridization was performed with PDGF-B chain cDNA probe and the results were quantitatively analyzed to measure the changes in PDGF-B chain mRNA expression in the lung tissue. The results showed that: (1) MP polymerase chain reaction (PCR) tests showed positive results in the bronchoalveolar lavage fluid (BALF) from all of the MP-infected rats (n = 4) while they were all negative in BALF from the control animals (n = 4, P < 0.05) and in BALF from those rats both infected with MP and, at the same time, treated with erythromycin (n = 4, P < 0.05). Bacterial cultures of the bronchial and lung tissue were negative in all three groups. The observation under a transmission electron microscope indicated that the interalveolar septa were widened with increased amount of collagen in the MP infected rats while there were no obvious abnormalities in the other two groups. (2) Strong positive expression of PDGF-B chain mRNA was found in the plasma of monocytes and macrophages located in the locally widened interalveolar septa and alveolar spaces in the lung tissue from the MP-infected animals with the integral optical densities being 37.42 +/- 9.05 (n = 4) which was significantly higher than the values of control group (0.42 +/- 0.08, n = 4, P < 0.01) and of the group with MP-infection plus erythromycin treatment (1.62 +/- 0.40, n = 4, P < 0.01). These results suggest that PDGF-BB may be involved in the process of the development of pulmonary interstitial fibrosis caused by the repeated MP infection. It may be an important growth factor for mediating the roles of monocytes and macrophages to promote the aggregation and proliferation of fibroblasts which can then secrete collagen in large quantity in the pulmonary interstitium. PMID- 10806850 TI - Experimental study on the correlation of nitric oxide with portal hypertensive enteropathy. AB - To explore the role of nitric oxide (NO) in the pathogenesis of portal hypertensive enteropathy (PHE), cirrhotic portal hypertension was induced in Wistar rats by subcutaneous administration of carbon tetrachloride. At the end of 12th week, NADPH-diaphorase staining was performed on ice frozen sections with the sample of fresh colonic tissues. NO synthase (NOS) activities and NOS mRNA expression of colonic tissues were investigated with chemoluminescence method and reverse transcription-polymerase chain reaction (RT-PCR), respectively. After NADPH-diaphorase histochemical staining, the computer image analysis and paired t test showed that NOS staining intensities of submucosal vascular endothelial cells and nerve fibers were all significantly higher in PHE group than those in normal group (P < 0.01 and P < 0.05, respectively), but the intensities of superficial epithelial cells were significantly lower than those of controls (P < 0.01). Chemoluminescence method demonstrated that general NOS activity of colonic mucosa was significantly higher in PHE group than that in control group (P < 0.01). Moreover, the degree of iNOS activity increase was greater than that of cNOS (104% vs 35%). RT-PCR revealed that NOS mRNA expressions were dramatically higher in PHE group than those in control group (P < 0.01). The results suggested that NO, with its property of vasodilatation, may be involved in the pathogenesis of vascular lesions of PHE in cirrhotic rats, and may also has something to do with mucosal lesions of colon in PHE. PMID- 10806851 TI - Changes of gastric nitric oxide in neonatal rats with hypoxic-ischemic encephalopathy. AB - In order to study the changes of nitric oxide (NO) in gastric wall in hypoxic neonatal rats, the activity of nitric oxide synthase (NOS) of gastric wall was measured and the NADPH-diaphorase biochemistry test was taken to show the distribution of NOS in the gastric wall of neonatal rats. The results showed that compared with normal rats, NOS had no significant changes in the acute hypoxic rats (P > 0.05). However, in hypoxic-ischemic encephalopathy (HIE) neonatal rats, the activity of NOS was markedly increased (P < 0.01), and the NOS positive reactive products were markedly increased in the gastric muscle, but no changes in the mucosal and submucosal layers were observed. The results suggest that the decreased gastric motivation and the gastric mucosal lesions caused by asphyxia are associated with the changes of NO in gastric wall. PMID- 10806852 TI - Effect of SJAMP on human platelet cytoplasmic Ca2+. AB - Using the method of dual-wavelength measurement of platelet [Ca2+]i and Fura-2 as the Ca2+ fluorophore probe, we measured the effect of acidic Mucopolysaccharide from Sticopus Japonicus Selenka (SJAMP) on platelet [Ca2+]i. The results showed that the most significant increase in platelets [Ca2+]i was seen when the concentration of SJAMP was 100 micrograms/ml and the elevation of normal platelet [Ca2+]i was 93.96 +/- 10.24 nmol/L (n = 10). In the presence of extracellular Ca2+ (1 mmol/L), the magnitude of platelet [Ca2+]i response to SJAMP was increased and the [Ca2+]i could reach 116.72 +/- 10.66 nmol/L (n = 10). On the other hand, the magnitude of increased platelet [Ca2+]i induced by SJAMP was smaller and the duration of [Ca2+]i reaching the highest level was longer when compared with other platelet aggregation agents. In the mean time, if platelets were first incubated with cyclooxygenase inhibitor, the rise of [Ca2+]i evoked by SJAMP was inhibited. The results indicated that the mechanism of the rise of [Ca2+]i induced by SJAMP might be dependent upon the generation of prostaglandin endoperoxides and(or) TXA2. PMID- 10806853 TI - Variations and clinical significance of coagulation and fibrinolysis parameters in patients with diabetes mellitus. AB - We observed the changes of parameters of coagulation and fibrinolytic system in order to understand the clinical implication of these variations in type II diabetic patients. Subjects consisted of 22 patients with type II diabetes mellitus and 25 healthy controls. Compared with the control, activated partial thromboplastin time, prothrombin time were shortened in the patients. The diabetic subjects also displayed higher levels of D-dimer, serum fibrin degradation products, median concentrations of fibrinogen (3.99 vs 2.96 g/L, P < 0.01) and von Willebrand factor (149% vs 87%, P < 0.01). Levels of antithrombin III activity or antigen were not different from control values. Simple linear regression analysis revealed a negative correlation between antithrombin III activity and fast blood glucose. Diabetic patients with vascular complications had significantly higher levels of fibrinogen and D-dimer than those without diabetic angiopathy. Our data demonstrated that patients with type II diabetes mellitus had a hypercoagulable state. We believed the activation of coagulation might contribute to the vascular complications in diabetics. PMID- 10806854 TI - Serum lipoprotein (a) concentration in patients with nephrotic syndrome and its clinical implication. AB - Serum lipoprotein(a) [Lp(a)] concentration was determined in 42 patients with primary nephrotic syndrome (NS) and the relationships between Lp (a) and plasma lipids, apolipoproteins, serum creatinine (Scr), albumin, urinary proteins (Upro) were also analyzed. The results showed that: (1) serum Lp(a) concentrations in the patients with NS were higher than those in healthy controls; (2) the levels of serum Lp(a) were correlated positively with total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), apolipoprotein B (Apo-B), Upros (Upro). It is concluded that the NS patients had the potential risk of suffering from coronary artery disease, glomerular sclerosis and thrombosis. The remission of NS may partially decrease the serum Lp(a) levels. Further studies are needed to explore the prevention and treatment of dislipedemia in patients with NS. PMID- 10806855 TI - Preventive effect of different dosage of recombinant human erythropoietin on anemia of premature infants. AB - To assess the efficacy and the optimum dose of recombinant human erythropoietin (rhEpo) on the anemia of premature, 45 preterm infants with a gestational age of less than 35 weeks and birth weight of less 1,800 g were randomly assigned to treatment group 1 (n = 15, receiving subcutaneous rhEpo 150 U/kg.time), treatment group 2 (n = 15, receiving 250 U/kg.time), three times a week for 6 weeks, and control group (n = 15, no treatment was given). All preterm infants received supplements of vitamin E (20 IU) and iron (20 mg) each day. Our results showed that postnatal decline of hemoglobin (Hb) and hematocrit (Hct) were lessened in the treatment groups, particularly in the group 2 and the differences were very significant (P < 0.0001 for all). Treated infants had significantly higher reticulocyte counts (Ret) (P < 0.0001 for all), but there was no significant difference between the two treatment groups (P > 0.05). Serum iron dropped significantly in the treatment groups as compared with control group (P < 0.01 for all), but no dose-dependent relationship was observed in treated infants (P > 0.05). After treatment, serum levels of erythropoietin was higher in group 2 than those in group 1 and control group (P < 0.0001, P < 0.01 and P < 0.05, respectively). There was no significant difference between group 1 and control group (P > 0.05). No side effects related to rhEpo therapy were observed. Our study suggested that rhEpo therapy stimulates endogenous erythropoiesis and enhances Ret, Hct and level of Hb in a dose-dependent manner in premature infants. The therapy is more efficient when given in higher dosages. PMID- 10806856 TI - Kawasaki disease on PDGF expression and VSMC proliferation. AB - The effect of serum of patients with Kawasaki disease (KD) on expression of platelet-derived growth factor (PDGF) B chain protein in vascular endothelial cells (VEC) was studied by immunocytochemical method. Meanwhile, the effects of the endothelial cell conditioned media (ECM) on expression of PDGF receptor mRNA in vascular smooth muscle cells (VSMC) and on cell cycle of VSMC were investigated by the methods of nucleic acid hybridization and flow cytometry (FCM). The results showed that the serum of patients with KD induced the expression of PDGF-B chain protein significantly. ECM significantly promoted the expression of PDGF receptor mRNA and induced the proliferation of VSMC. These data suggest that the activation of PDGF-PDGF receptor may play a role in the pathogenesis of coronary complication of KD. PMID- 10806857 TI - Tumor necrosis factor alpha, interleukin-6, interleukin-8, and interferon alpha in children with viral hepatitis. AB - To investigate the cellular immunological changes in children with viral hepatitis, interleukin 6 (IL-6), interleukin 8 (IL-8), interferon alpha (IFN alpha), and tumor necrosis factor alpha (TNF alpha) in supernatant of cultured peripheral blood mononuclear cells (PBMCs) of 49 children with hepatitis A, B or C were measured. The levels of IL-6, IL-8, TNF alpha in PBMCs of the 3 viral hepatitis groups were increased and the level of IFN alpha decreased as compared with those of normal control group. But there were no significant differences among the 3 viral hepatitis groups. It was concluded that cellular immunological disorders were related to the onset and the induced damage of the viral hepatitis in children. PMID- 10806858 TI - A prospective study on HGV infection after transfusion in pediatric hematological children. AB - In order to delineate the infectious status of HGV in pediatric hematological patients and its clinical features, 38 children were studied for HGV by reverse transcription nested polymerase chain reaction (RT-nPCR). The overall positive rate was 23.68%; the positive rate in 1-10 time transfusion recipients (mean 3.26 times) was 3.70% while in over 10 time transfusion recipients (mean 20.09 times) was 72.73%. A statistically significant difference was found between the two groups (P < 0.001). All HGV positive children showed diminished immunity and most were single HGV infection with no clinical and biochemical evidences of acute hepatitis. Our results suggest that HGV is one of the main causes of post transfusion hepatic virus infection and the prevalence is related to transfusion times. Hematological malignancy maybe the most susceptible confluence of HGV. PMID- 10806859 TI - Endovascular stents in arterial injury after radiotherapy. AB - We evaluated preliminary success and patency of stenting for the treatment of radiation-induced arterial diseases. Thirteen stents were placed in 8 patients to treat occlusion (n = 3), aneurysm (n = 1), residual stenosis (n = 2), multiple stenoses (n = 1), and delayed restenosis after previous balloon angioplasty (n = 1). Interventional procedure was successfully performed in 8 patients for their arterial lesions after radiotherapy. Six patients underwent interventional procedure once or twice. Two patients underwent PTA 4 times. Five of these patients demonstrated primary patency with relief of clinical symptoms with a mean follow-up of 2 years (range: 8-60 months). Clinical improvement was noted in the other patients. Our results suggest that stent placement by single or multiple techniques may have immediate effect on arterial lesions caused by radiation and can be considered as a therapeutic option of choice in these cases. PMID- 10806860 TI - Biodegradation of tricalcium phosphate ceramics by osteoclasts. AB - Biodegradation of tricalcium phosphate (TCP) ceramics was observed through mixed culture of osteoclasts and TCP discs in vitro in this study. Osteoclasts were isolated from newborn SD rat's marrow of long bone and cultured on TCP discs. The culture terminated at the 48th h and 96th h respectively. Under an inverted microscope, the osteoclasts imparted round or oval body with multinuclear and many thin processes. These cells were positively stained for tartrate-resistance acid phosphatase (TRAP). Scanning electron microscope showed that many resorption lacunae on TCP disc surface and their diameters were smaller than 20 microns. Osteoclasts were located in the lacunae. At the 96th h, the resorption lacunae become larger and osteoclasts showed degeneration. It is suggested that osteoclasts possess ability to re-absorb TCP ceramics under in vitro culturing condition. PMID- 10806861 TI - Embolization of tumors and vascular lesions in the oral and maxillofacial regions. AB - The embolization was used for the treatment of 10 patients with tumors and vascular lesions in the oral and maxillofacial region. The patients were diagnosed as having vascular deformity, hemangioma, hemangiofibroma and chemodectoma. The selective and superselective target artery embolization were conducted and the satisfactory results were achieved. The procedures, effects and precautions of the embolic therapy were also discussed. PMID- 10806862 TI - [Structure and function of MHC: molecular mechanisms for antigen processing and antigen presentation to T cells]. PMID- 10806863 TI - [Involvement of membrane phospholipid metabolism in cell motility]. PMID- 10806864 TI - [Molecular biology concerning to the honeybee sociality: how can we approach the evolution of insect sociality?]. PMID- 10806865 TI - [Functional structures in the ribosome revealed by a crystallographic analysis]. PMID- 10806866 TI - [X-ray crystal structure analysis of ribosome]. PMID- 10806867 TI - [Creation of polymeric micelles as devise for drug delivery inspired by structural properties of natural viruses]. PMID- 10806868 TI - [Mass spectrometry of protein (2): Time-of-flight mass spectrometer and its application]. PMID- 10806869 TI - [Synchrotron radiation facilities for structure biology in the PF and SPring-8]. PMID- 10806870 TI - [New extra tax for the sick]. PMID- 10806871 TI - [Ward units]. PMID- 10806872 TI - [Radiation--only in connection with cancer?]. PMID- 10806873 TI - [Injuries connected to violence]. PMID- 10806874 TI - [Good news on coronary surgery]. PMID- 10806875 TI - [Coronary surgery at the Heart Center in Oslo 1989-98]. AB - BACKGROUND: Coronary artery bypass grafting was performed in 5,658 consecutive patients during the period 1989-1998. Due to changes over time, both in patients' risk profile and surgical strategies, a review was undertaken to study trends and results after coronary artery bypass surgery. MATERIAL AND METHODS: Our database includes more than 160 variables per patient, covering preoperative risk factors, catheterization data, operative and postoperative results. These data form the basis for analysis over time. RESULTS: Median age increased for both genders, from 58 years to 64 years for males and 62.5 years to 69 years for females. The female proportion increased from 12.8% to 19.8%. A high operative risk profile was registered in 23.7% in 1989 compared to 61.8% in 1998. Heparin-coated extracorporeal equipment and blood cardioplegia were gradually introduced in routine practice. Despite higher age and operative risk profile the morbidity and hospital mortality (0.41% overall) remained nearly unchanged. INTERPRETATION: Due to continuous improvement of technical equipment and treatment strategies, coronary artery bypass surgery represents a safe option for both high and low risk patients. PMID- 10806876 TI - [Check list for the use of thrombolytics and other drugs in acute myocardial infarction]. AB - MATERIALS AND METHODS: Data on all patients with acute myocardial infarction who were treated in Nordland County Hospital in a six-month period in 1996 were analyzed retrospectively (137 patients). After the introduction of checklists for the treatment of such patients, we did a prospective six-month registration in 1997 (111 patients) in order to find out whether treatment and delay times had improved. RESULTS: The proportion of patients who received thrombolytic treatment did not change (28% in 1996 as compared to 25% in 1997). The in-hospital delay time before treatment did not differ before and after the introduction of a check list (approximately 40 minutes in both periods). There was an increase in the use of intravenous beta blockers and aspirin. INTERPRETATION: The percentage of patients with acute myocardial infarction receiving thrombolytics in our hospital does not differ substantially from that of other hospitals in Norway. PMID- 10806877 TI - [Choanal atresia--experience with early surgery]. AB - INTRODUCTION: Prior to 1982, patients in our department with choanalatresia were operated by the transpalatinal approach. To eliminate the risk of insufficient transpalatine growth, only older children were operated. MATERIAL AND METHODS: From 1982 we have chosen to operate as soon as symptoms occurred, usually at an earlier age. Therefore, we chose the transnasal approach in 32 patients. RESULTS: Of 14 patients with bilateral choanalatresia, nine were operated before two months of age. In nine patients we took care to remove some of the posterior part of vomer. Four of them have been symptom free for more than a year, an additional two symptom free but with an observation period of less than a year, and three have renewed symptoms. Of 18 patients with unilateral atresia, 11 were operated before seven years of age, seven of them successfully. Of the seven operated after the age of seven, six are symptom free. INTERPRETATIONS: We believe that these results justify continuing with transnasal operations as soon as symptoms are distressing, regardless of age. PMID- 10806878 TI - [Endophthalmitis]. AB - Endophthalmitis is a rare, but serious complication of intraocular surgery and penetrating injury, and is occasionally seen as a consequence of microbial spread either systemically or from a local focus of infection. Data from 34 patients with endophthalmitis at Haukeland University Hospital from 1988-98 were reviewed. The number of patients with endophthalmitis per year was 0-4, except for 1998 when 11 patients were diagnosed. Six patients had been treated with antibiotics prior to admission and were culture negative. 29 isolates were obtained form the other 27 patients. Pneumococci were identified in eight. Predisposing factors included ocular surgery, penetrating trauma, and periocular or systemic infections. All patients were treated with local and systemic antibiotics (from 1995 with a standardised combination of vancomycin and ceftazidime). Vitrectomy was performed in 22 patients. The visual function was perception of light or worse in six patients prior to the infection, and was the final outcome in 22 patients after the infection. The prognosis was dependent on predisposing factors and the infecting organism. Vitrectomy did not seem to improve the results. This series illustrates the clinical picture of endophthalmitis and current options for treatment. No definite explanation for the increased number of cases seen in 1998 was found. PMID- 10806879 TI - [Diffusion-weighted magnetic resonance tomography--a new instrument in the diagnosis of Creutzfeldt-Jakob disease]. AB - Creutzfeldt-Jakob's disease (CJD) is characterised by rapidly progressive dementia, ataxia, myoclonus, and several other neurological deficits. It generally affects older adults and occurs in sporadic, genetic and iatrogenic forms. Death occurs usually within one year after onset of the disease. The diagnosis is based on clinical criteria, neurophysiological and radiological findings, and confirmed by postmortal histopathology. During the last two years several cases of CJD have been reported with diffusion-weighted magnetic resonance imaging (MR) abnormalities, represented by increased signal intensity indicating reduced diffusion in basal ganglia and/or cortex cerebri. These abnormalities seem to be characteristic of CJD. We report a case of CJD in a 54 year-old woman who developed vertigo, nystagmus, ataxia, myoclonus, and dementia over a period of eight months. Diffusion-weighted magnetic resonance imaging showed increased signal intensity in corpus striatum and gyrus cinguli. The diagnosis was postmortally confirmed with histopathology. PMID- 10806880 TI - [Medical rehabilitation in nursing home--effects on functional outcome and use of the services]. AB - BACKGROUND: Improved functioning is a primary objective in nursing homes. MATERIAL AND METHODS: A total of 146 patients participated in the study, 116 for rehabilitation and 30 for respite care. 122 were admitted from home, 23 from hospital and one from a full-time care facility. The mean age was 78. The multi disciplinary team consisted of a geriatrician, registered nurses and auxiliary nurses, an occupational therapist, a physical therapist and a speech therapist. Assessments were made before admission and about two months after discharge. The mean number of days in the nursing home was 35 (28-42 days). RESULTS: A total of 141 were able to resume living at home, whereas five were discharged to permanent care in a nursing home. At the follow-up, gait was improved for 23, was unchanged for 121, and had deteriorated in two (p < 0.001). Of the 21 patients who had been confined to wheelchairs or were bedridden, eight were now able to walk. Regarding change in ADL functions, only personal hygiene reached statistical significance (p < 0.05). Perceived global coping was improved for rehabilitation patients (p = 0.05). No significant changes in mental functioning or in the use of nursing services were found. The average number of hours of home help was reduced from 3.3 to 2.7 a week (p = 0.01). INTERPRETATIONS: The most notable effects were improved mobility and decreased dependence on home-help services. PMID- 10806881 TI - [Long-term follow-up of patients with congenital heart defects]. AB - There are about 500 new cases of congenital heart disease per year in Norway. Modern diagnostic skills, surgical techniques and follow-up programs have contributed to higher survival rates among patients. Based on international experience, 85-90 per cent of these children will survive into adulthood. Half will suffer from conditions which should be followed up by cardiologists. This article is based upon recommendations on long-term follow-up of patients with congenital heart disease. PMID- 10806882 TI - [Magnetic resonance tomography with endorectal coil for examination of prostate and seminal vesicles]. AB - MRI with an endorectal coil gives images of the prostate gland and seminal vesicles of higher quality than any other imaging modality. The use of an endorectal coil enables us to sample stronger radiofrequency signals from the prostate gland and seminal vesicles compared to a whole-body coil. This improves the contrast resolution in MR images and may give a higher spatial resolution by use of thin slices and a smaller slice gap. Intravenous MRI contrast is not used in our standard procedure. At Haukeland University Hospital, Norway, MRI with an endorectal coil is presently used as a tool for solving special problems in relation to detection and staging of prostate cancer, for example in patients with an abnormal s-PSA and negative sextant biopsies from the prostate gland. Furthermore, MRI with an endorectal coil may be used for the diagnosis of some benign diseases. The specific role of MRI with an endorectal coil in clinical use is not clarified; further research is needed. This article gives a short overview of technical aspects, imaging protocols, image reading, indications and diagnostic criteria for some diseases. PMID- 10806883 TI - [The use of cottage hospitals' beds in Nordkapp]. AB - An analysis was made of 414 admissions during a one-year period to three general practitioner beds. Within the first day, 20% of the patients were discharged, while 22% were transferred to the main hospital after examination and primary treatment. 58% stayed more than one day. The mean stay was 5.0 days. The three major groups of medical conditions according to the International Classification of Primary Care were cardiovascular diseases, diseases of the musculoskeletal system and diseases of the lungs and the respiratory tract. Patients > or = 60 years of age constituted 55.8% of the total, taking up 80.6% of total bed days. In the age group > or = 80 years of age, there were three admissions for every five inhabitants, while two in five had one or more hospital stay. 65% of all patients (15.7%) had two stays or more, taking up a total of 60.5% of total bed days. Using the general practitioner beds as a low threshold service proved especially useful with patients suffering from heart failure, asthma or chronic obstructive lung disease. The beds had a key function in the rehabilitation of the elderly, in the care of cancer patients, and in terminal care. They were of basic importance to the organisation of daily emergencies. PMID- 10806884 TI - [The cottage hospital model, a key to better cooperation in health care--let the cottage hospital survive!]. AB - The cottage hospital model may be defined as an intermediary service between primary care and the general hospital. On the basis of experience and studies from Finnmark county, the northern-most county in Norway, this article makes a case for a revival of the cottage hospitals. They may improve comprehensive patient care and cooperation between care levels, to the benefit of groups of patients who often are in a squeeze between care levels: the elderly, the chronically ill, and the severely ill and dying patients. The cottage hospitals may also contribute to strengthening the chain of service in acute medicine. The professional challenges of work in a cottage hospital may attract practitioners to primary health care. We suggest that 1% of the funds set aside for ongoing national programmes for the elderly, in cancer care and mental illnesses are used for cottage hospital beds, as this may contribute to increasing the viability of these programmes. The extra cost upgrading 1,000 of a total of 27,000 nursing home beds in Norway to cottage hospital standard is estimated to be modest. PMID- 10806885 TI - [Can radiotherapy reduce the frequency of restenosis after coronary angioplasty?]. AB - Today, percutaneous transluminal coronary angioplasty (PTCA) is the most important treatment modality for coronary artery disease. However, restenosis occurs in 20-40% of the vessels in spite of the use of stents. Intravascular radiation therapy has reduced the frequency of restenosis in both animal studies and clinical trials. Recent randomized trials have shown a reduction in the order of 30% after irradiation. A number of techniques which use different sources of radiation (beta vs. gamma radiation) are under investigation. This review gives and update of intravascular radiation therapy, including a discussion of ongoing trials. There is a great need, also in Norway, for treatment modalities that reduce the frequency of restenosis after PTCA. It would seem imperative that we start a discussion of whether and when intravascular radiation therapy should be available in Norway. Implementation requires planning on a national level with regard to investment in equipment and training. PMID- 10806886 TI - [Injuries connected to violence--an analysis of data from the injury registry]. AB - BACKGROUND: Injury purposely inflicted by other persons is a potentially significant public health problem in Norway, and the size of the problem has received little attention. MATERIAL AND METHODS: We used data from the Norwegian National Injury Sample Registry from 1990-1997 and investigated occurrence of such injuries. RESULTS: A total of 9,649 injuries were included in this study, or 3.8 injuries per 1,000 person-years. Of these, 16% (0.6 per 1,000 person-years) required inpatient treatment. Injuries due to violence represented 2.4% of all injuries among women and 5.5% of all injuries among men. The incidence rate for men and women was highest in the age group 15-24 years. The incidence rate among men in this age group was 4-5 times the rate among women. Domestic violence caused at least 39% of injuries among women. Weekend injuries were overrepresented; about 64% of the injuries occurred from Friday to Sunday. INTERPRETATION: Figures represented in this study reveal only a portion of the problem of violent injuries. Spouse abuse, mental abuse, maltreatment, and sexual abuse without a physical injury are not represented in our data. PMID- 10806888 TI - [The cause of death registry and research]. PMID- 10806887 TI - [Late sequelae of violence seen from victims' perspective]. AB - Hospital accident and emergency (A&E) departments frequently treat victims of interpersonal violence. This study of 1,451 victims treated at the A&E departments of Aarhus University Hospital during a one-year period (1993-94) aims at illuminating some of the sequelae of victims. A questionnaire survey was carried out two years later to obtain information on medical after-treatment, the extent of sick leaves, and the victims' personal costs. One in two assault victims needed further medical treatment. 7.5% were admitted to hospital and 20% were referred to the out-patient clinic in the hospital or to follow-up treatment in general practice. The median sick leave period was eight days. Two years after the violence took place, 21.8% of all assault victims claimed that they had cosmetic sequelae, 14.7% had persistent pain. 38.1% said they were afraid to enter certain neighbourhoods or places; 41.2% had frequently or occasionally thought about the assault. The study has revealed some aspects of the personal costs, discomfort and trouble that victims of violence experience. PMID- 10806889 TI - [European guidelines for health care in hereditary breast cancer]. PMID- 10806890 TI - [How to achieve an optimal intake of vitamin D in children]. PMID- 10806891 TI - [Cholesterol reducing health food products]. PMID- 10806892 TI - [Health care reforms--progress or a patchwork?]. PMID- 10806893 TI - [General practice in crisis]. PMID- 10806894 TI - [Investigation of syncope]. PMID- 10806895 TI - [Sjogren's syndrome]. PMID- 10806896 TI - [Antibiotic prophylaxis in Cesarean section]. PMID- 10806897 TI - [Insufficient help to victims of violence]. PMID- 10806898 TI - [How much money are we willing to spend on health services?]. AB - BACKGROUND: Like most other countries, Norway spends increasing sums of money on health care. The purpose of this study is to elicit people's views on whether society should spend more, and if so, their willingness to contribute to the financing of an expanded health service. MATERIAL AND METHODS: A random sample of the Norwegian population (2,089) were approached regarding a questionnaire study. 716 returned completed questionnaires (34%). Different versions of a questionnaire were used on three sub-samples in order to analyze the extent to which the distribution of answers depends on the wording of the questions. Variation in answers are sought explained by sociodemographic variables and political preferences. RESULTS: 70-80% held that society should spend more on health care. Their willingness to pay more in terms of "earmarked health care taxes" varied between the sub-samples. The mean annual figures were between NOK 1,314 and NOK 1,972. The proportions not willing to pay more varied between 39% and 46%. INTERPRETATION: It appears to be wide support for the idea that society should spend more on health care, but limited support for the idea of having to finance the desired expansion. One should be very cautious to generalize from preference surveys of this kind, because answers depend on the wording of the questions. PMID- 10806899 TI - [Size of the patient list and quality of the general practice]. AB - BACKGROUND: A list patient system will be introduced in Norway from 2001. In England and Holland, countries with long experience with this system, the average consultation is much shorter than in the present Norwegian system. Research reports indicate a significant association between longer consultations and patient satisfaction with service as well as quality. The challenge in these countries has been to find mechanisms that reduce list size in order to enhance the quality of care. MATERIAL AND METHODS: On the basis of the author's own data and a summary of relevant literature, a model for estimating average list size in Norway is developed. RESULTS: The model shows the impact of various time consuming factors in general practice on the workload associated with a patient list. INTERPRETATION: The article describes some objective criteria for calculating a list size which corresponds to a full normal medical man-year, with due consideration for quality of care. PMID- 10806900 TI - [Six years with the general practice system--what is the opinion of general practitioners?]. AB - BACKGROUND: In 1993 a list patient system was introduced in four Norwegian municipalities: Trondheim, Tromso, Lillehammer and Asnes. MATERIAL AND METHODS: Questionnaire surveys were conducted among all GPs in these four municipalities in 1996 and 1999 for the purpose of examining changes in their assessment of the list patient system. RESULTS: The number of GPs generally satisfied with the list patient system declined over the three-year period. Doctors found their workload heavier in 1999 than in 1996, but their confidence in the list patient system as a tool for enhancing quality in general practice did not change over the period. GPs with too long patient lists were the most dissatisfied. The decline in confidence in the new system was strongest in Trondheim, the largest of the four municipalities and that in which the average patient list is longest. In Tromso, the second largest municipality, with moderate to average patient lists, there were no changes in GP attitudes. INTERPRETATION: The study demonstrates some important consequences of a shortage of medical manpower in a list patient system. Local authorities will have to deal with these challenges. PMID- 10806901 TI - [Symptoms in women with Sjogren's syndrome]. AB - More knowledge about symptom experience is needed for diagnosing Sjogren's disease at an early stage of disease development. We have performed a qualitative study based on group interview with eight women with Sjogren's disease. The women presented with several different symptoms. They all described weariness, fatigue, sicca symptoms, various pains, hypersensitivity and organ manifestations in the same manner. Some of the symptoms had a dramatic impact on their life activities. The study revealed new descriptions of previously known symptoms and descriptions of previously unfamiliar symptoms. The results of the study may supplement the existing criteria of Sjogren's disease and provide new hypotheses about pathogenetic mechanisms related to chest pain, abdominal pain and facial pain in patients with Sjogren's disease. PMID- 10806902 TI - [Arthrocentesis in rheumatology practice]. AB - This study of Norwegian rheumatologists' use of intraarticular steroid injections is based on a survey among members of the Norwegian Society for Rheumatology. 79% of the members responded, i.e. 108 rheumatologists. 69 reported having used intraarticular steroid injections in any joint during the last week, a total of 637 times. There have been no previous studies on this subject in Norway. The results show that Norwegian rheumatologists consider intraarticular steroid injections a very effective treatment. Only 9% reported that they had seen side effects over the last 12 months (a total of 51 side effects), of which post injection pain and subcutaneous atrophy were the most common. There were no reports of septic arthritis. Almost all side effects were considered not serious. PMID- 10806903 TI - [Use of tilt test in the investigation of syncope]. AB - INTRODUCTION: In evaluating syncope, a 45-60 minutes head-up tilt test at 60 degrees with or without pharmacological stimulation is often used. MATERIAL AND METHODS: We report our experience with a faster, ten minute tilt test at 80 degrees with retilting during isoprenaline infusion, on 15 healthy volunteers and 27 patients who subsequently were followed up during a period of 1-3 1/2 years. RESULTS: One volunteer had a positive response (presyncope), while seven of the 27 patients reproduced their clinical symptoms. Among the 20 patients with normal test responses, the clinical course revealed the diagnosis in 11. For the remaining nine patients a definite diagnosis has not been made, but they have not experienced further syncopal episodes. INTERPRETATION: This study demonstrates that the abbreviated tilt test is useful for evaluating recurrent vasovagal and orthostatic syncope. PMID- 10806904 TI - [Investigation of patients with syncope]. PMID- 10806905 TI - [Heredity and immunology in Sjogren's syndrome]. AB - BACKGROUND: Over the next 3-5 years, the rapid progress in genomic research will enable the discovery of many genes associated with the more common diseases. An example of such a common disease is the rheumatic disorder Sjogren's syndrome, an autoimmune disease. A more precise genetic explanation of the mechanism leading to Sjogren's syndrome remains to be given. MATERIAL AND METHODS: One way of investigating the disease related genes in such complex polygenic diseases is to perform linkage studies in families with two or more affected. Another possibility is to conduct association studies on trios (parents and affected child), case control studies, or other experimental designs. In association studies one is testing if an allele is significantly elevated among patients compared to controls, while in linkage analyses one finds subchromosomal regions that are significantly more often inherited by patients than by healthy family members. RESULTS: The most well defined genetic association in Sjogren's syndrome is currently related to different HLA alleles and their association with anti Ro/SSA and anti-La/SSB autoantibodies. Additional genetic studies focusing on non HLA regions are under way. INTERPRETATION: Increased genetic knowledge would allow optimisation of the diagnostic criteria as well as development of new and more effective treatment for Sjogren's syndrome, which causes substantial suffering for a large group of patients. PMID- 10806906 TI - [Use of angiotensin II receptor blockaders in heart failure]. AB - Angiotensin converting enzyme (ACE) inhibitors and diuretics represent the first line of therapy in patients with symptomatic heart failure. Inhibition of angiotensin II production is, however, incomplete with ACE inhibitors, due to non ACE dependent conversion pathways. Moreover, some patients are intolerant to ACE inhibitors due to side effects or renal insufficiency. Angiotensin II receptor blockers may be an alternative to, or an additional treatment in heart failure. Preliminary studies comparing the angiotensin II receptor blocker losartan with placebo have demonstrated improved haemodynamic parameters, reduced hospitalisation and mortality in patients with heart failure. Reduced morbidity and mortality have also been found with losartan treatment, as compared to the ACE inhibitor captopril. This paper discusses the role of angiotensin II receptor blockers in the treatment of heart failure. Some results from published studies and a short description of ongoing trials are presented. PMID- 10806907 TI - [Controversies in surgical treatment of advanced ovarian cancer]. AB - Cytoreductive surgery has traditionally been regarded as a cornerstone in the primary treatment of advanced ovarian cancer. Both five year survival and median survival are better for patients with small residual masses. Despite many similar reports showing the prognostic significance of postoperative residual tumour, the survival benefits of cytoreductive surgery still remain scientifically unproven and controversial. There have been no prospective controlled clinical trials. The question remains as to whether the observed survival benefits for patients subjected to primary cytoreductive surgery are an effect of surgery skills or tumour biology. The proponents of tumour biology claim that cytoreductive surgery is a selective procedure and that patients with better prognosis are selected. Therefore a randomized study between primary cytoreduction and neoadjuvant chemotherapy in patients that cannot be optimally cytoreduced seems warranted, though one problem with such a study is how to select eligible patients. During chemotherapy and after relapse several types of operations are used in ovarian cancer: secondary cytoreductive surgery, interval cytoreductive surgery, second look surgery and palliative secondary surgery. So far interval cytoreductive surgery during chemotherapy is the only type of operation which in a prospective randomized study showed significant improvement in long-term survival. This paper discusses indirect evidence in the literature in support of or in contradiction to the primary debulking hypotheses and also indications and impact of surgical procedures during chemotherapy. PMID- 10806908 TI - [Magnetic resonance tomography in sinus tarsi syndrome]. AB - The sinus tarsi is a cone-shaped anatomical space in the hind foot that is bounded by the talus and calcaneus, the talonavicular and posterior sub-talar joint, and is continuous with the tarsal canal medially. The space courses from posteromedial to anterolateral, with the larger portion of the cone laterally situated. The sinus tarsi consists of fatty tissue, arterial anastomoses, joint recesses posteriorly, nerve endings, and five ligaments. These include the lateral, intermediate, medial roots of the inferior extensor retinaculum, the cervical ligament, and the interosseous ligament. The sinus syndrome is characterized by lateral foot pain, focal pain to palpation over the tarsal sinus, and hind foot instability. This chronic disease is related to a history of inversion trauma in the majority of patients. Magnetic resonance imaging (MRI) may be used in the evaluation of abnormalities associated with the sinus tarsi syndrome. PMID- 10806909 TI - [Patients in general practice who experienced violence or threats from someone they knew well]. AB - INTRODUCTION: Having been exposed to threats or violence often influences health. We explored the association between such experiences and sociodemographic data among patients in general practice. MATERIAL AND METHODS: A questionnaire study was conducted. 89 doctors and their patients during one workday were the informants. The patients completed the questionnaire at home. RESULTS: 1,401 consultations were included; 1,217 (87%) patients responded. 14% of the patients said that they experienced violence or threats from someone they knew well, 2% were uncertain. 38% of adult patients in single-parent household had such experiences. The prevalence was also higher among patients with a difficult housing situation, in need of social welfare, who had been unemployed for more than six months, or assessed themselves as more than 50% permanently disabled. 6% of patients 60 years or older had experienced of violence or threats. INTERPRETATION: In general practice, patients with social problems or living in single-parent households more often have experience of violence or threats. Comparatively few elderly people report such experiences. PMID- 10806911 TI - [Thatcher's health care reform--what happened?]. PMID- 10806910 TI - [Tooth and jaw injuries following violence--diagnosis and treatment in emergency department]. AB - Research over the last decade has confirmed that facial injuries caused by violence occur most frequently among young males, often under the influence of alcohol, and less frequently in women and children following domestic violence. Evidence of injuries may be used in court or in claims for compensation, and must be carefully documented. Facial asymmetry, deviation of the mandible in opening, altered occlusion and/or abrasions under the chin may be indications of jaw fractures; such patients are referred to oral surgery units for further treatment. Lacerations and ulceration in the oral mucous membrane should be treated as skin wounds. A fractured tooth should be referred to a general dental practitioner. Luxated teeth should be carefully replaced and immediately referred to a dentist for fixation. Completely avulsed teeth should immediately be replaced, preferably at the site of the accident, or immediately upon arrival at the clinic. Prompt referral to a dental practitioner for follow-up treatment is essential for the success of replantations. Additional documentation of the injury may be provided by photographs, radiographs and drawings. The letter of referral to the dentist should contain the following information: data on the patient, the circumstances, clinical findings, treatment performed and prescriptions given, together with advice given to the patient. PMID- 10806912 TI - [Medical genetics--knowledge with many challenges in the new century]. PMID- 10806913 TI - [Antibiotic prophylaxis in Cesarean section]. PMID- 10806914 TI - [Danish experiences with patient lists in the general practice system--chaos or flexibility?]. PMID- 10806915 TI - [Genetic changes and clinical management in familial hypertrophic cardiomyopathy]. AB - Hypertrophic cardiomyopathy (HCM) is phenotypically and genotypically heterogeneous disease of heart. Nine chromosomal loci responsible for this condition have been identified: beta-myosin heavy chain, essential and regulatory myosin light chains, troponin T and I subunits, alpha-tropomosin, cardiac myosin binding protein C, cardiac actin and titin. These genes code for proteins involved in the contraction mechanism or in the control of contraction, therefore HCM has been classified as a disease of cardiac sarcomere. Over 107 mutations have been identified. More then half of them have been detected in the beta myosin heavy chain gene (beta-MHC). Some mutations in beta-MHC gene are associated with a benign prognosis, other are associated with high incidence of sudden cardiac death (SCD) and severe hypertrophy. Mutations in myosin binding protein C are associated with mild, delayed expression of cardiac hypertrophy and benign prognosis. Mutations in cardiac troponinT are associated with a mild degree of hypertrophy but a high incidence of SCD. Study of genes responsible for HCM will assume role in the context of clinical management of HCM, in particular regarding diagnosis and prognosis patients and families with HCM. PMID- 10806916 TI - [Treatment of solitary toxic thyroid nodules with the use of percutaneous ethanol injections]. AB - Twenty women aged 18-76 years (mean 42 +/- 23 years) with solitary benign toxic thyroid nodule were treated with intranodular percutaneous 96%-ethanol injection, performed under ultrasound guidance. 1 ml of ethanol was instilled per 1 cm3 of nodule tissue. 2-5 (mean 3.1) sessions were conducted every month. The early follow-up evaluation of the results (the evaluation of nodule size by means of clinical and ultrasound examination) was performed in 6 months, and the long-term one in 2 years after last injection. A morphological evaluation was conducted according to the following scale: 1--nodule disappearance, 2--cicatricial nodule (solid structure and less than 0.5 cm in diameter), 3--nodule smaller by [symbol: see text] 50% of its initial dimensions, 4--nodule smaller by < 50% of its initial dimensions. A functional evaluation was carried out according to the following scale: I--permanent euthyroidism, II--permanent decrease of hyperthyroidism, III--no functional effect. A final follow-up evaluation revealed: nodule disappearance in 2 (10%) patients (Group 1), cicatricial nodule in 8 (40%) patients (Group 2), nodules smaller by [symbol: see text] 50% of its initial dimensions in 2 (10%) patients (Group 3) and nodules smaller by < 50% of initial dimensions in 8 (40%) patients (Group 4). The results in Group 1 and 2 were defined as good (50%), in Group 3 as satisfactory (10%) and in Group 4 as bad; and I--permanent euthyroidism in 16 (80%) patients, II--permanent decrease of hyperthyroidism in 1 (5%) patient, III--no functional effect in 3 (15%) patients. 2 patients in whom new nodules appeared and 1 patient with bad morphological and no functional treatment effects were operated on. A histological examination revealed haemorrhagic necrosis surrounded by fibrous tissue in removed nodules. The following complications were observed: 1) pain during and after the injection in 15 (75%) patients, 2) subfebrile body temperature in 3 (15%) patients and fever up to 39 degrees C on the 1-st day after the injection in 1 (5%) patient, 3) local ecchymosis in 1 (5%) patient, 4) temporary dysphonia in 1 (5%) patient. It seems that percutaneous ethanol injection treatment can be an useful treatment method of solitary benign toxic thyroid nodules in patients who do not give their consent to surgical or 131I treatment or with contraindications to an operation or radioiodine therapy. PMID- 10806917 TI - [Trichological examinations in women suffering from diabetes mellitus]. AB - The lack of data on the process of alopecia in women suffering from diabetes mellitus made us undertake research in this area. The aim of this paper was the assessment of the state of head hair in trichological and clinical examinations, and on the basis of questionnaire. 50 women (age 44-82 years) were included in the study. Alopecia in women with diabetes mellitus is diffuse, located on the apex of the head and basic hair loss lies in telogenic pathomechanism. The highest percentage of telogenic hair is found in women treated with biguanides, and the lowest one in female patients taking insulin. PMID- 10806918 TI - [Efficacy of the impulse magnetic field therapy in patients after the outer drainage of abdominal cysts diagnosed with the use of isotopic and ultrasound techniques]. AB - 154 patients with pancreatic, renal or hepatic cysts have been treated with the outer drainage under USG monitoring. Because of cysts' contact with tracts penetrating to an organ or its lack, the patients have been divided into two groups. Directly after the puncture all the patients have undergone the high frequency impulse magnetic field therapy. A distinct, positive treatment effect has been found in patients who have gone through not only drainage, but also magnetic field. PMID- 10806919 TI - [The usefulness of ultrasonography in the diagnosis of pain caused by extensor knee apparatus' lesions]. AB - The aim of this work was to present our experience in using ultrasonography in the diagnostics of knee pain. In spite of some limitations of this method of examination it can be successfully used to obtain more accurate diagnosis of the extensor knee apparatus lesions. PMID- 10806920 TI - [Erythema multiforme in children versus Stevens-Johnson syndrome]. AB - 30 children treated between 1984-98 for erythema multiforme (versus SJS) in Department of Allergology at the Children's Memorial Health Institute in Warsaw. 25 children suffered from erythema multiforme minor and 5 suffered from Stevens Johnson Syndrome. Before symptoms of the illness 13 children had been treated with antibiotics and 27 children demonstrated different symptoms of infections, mostly viral. Drug therapies and viral or bacterial infections probably induce symptoms in patients susceptible to erythema multiforme or severe versus Stevens Johnson Syndrome. CONCLUSION: 1. Erythema multiforme (EM) rather seldom occurs in children. The Most commonly its course is benign (EM minor). In another type of erythema multiforme--Stevens-Johnson Syndrome (EM major), in which mucous membranes are involved, the course of disease and the prognosis are always severe. 2. Both viral and bacterial infections as well as administered drugs play the important role in the etiopathogenesis of erythema multiforme. 3. The treatment of erythema multiforme in children is symptomatic (general and local). In Stevens-Johnson Syndrome early administration of glicocorticoids is recommended. Children with bacterial infection (contagion, contamination) should be treated with antibiotic. PMID- 10806921 TI - [The application of biodegradable anastomosis ring Valtrac for intestinal anastomosis]. AB - In 1985 a biofragmentable anastomosis ring (BAR) Valtrac was introduced for intestinal anastomoses. From August 1994 trough March 1998, 49 intestinal anastomoses were performed in 44 patients: there were 28 jejunoileostomies, 8 colocolostomies and 3 gastrojejunostomies. In 5 patients after total gastrectomy, two anastomoses with the use of Valtrac ring were made during the same operation. The patient group consisted of 26 women and 18 men, aged 14 to 81 years (mean age: 54 years); there were 13 emergency and 31 elective procedures. The reasons for the operations were: cancer--35 cases, pancreatic cyst drainage--4 cases and reconstructive procedures in the digestive tract--5 cases. First intestinal gas passage and defecation were monitored in all patients; control X-ray was performed on the 10th postoperative day. No mortality nor serious complications were observed in the postoperative period. In an 81-year old patient, an inconsiderable leakage was found at the anastomotic site. This was successfully managed in the second operation. CONCLUSIONS: 1. Valtrac is an alternative method for intestinal anastomoses. 2. The anastomotic technique is atraumatic and easy even for an unexperienced surgeon. PMID- 10806922 TI - [The modified laparostomy with the use of ultrasound dissector and garamycin sponge in the treatment of complicated acute pancreatitis]. AB - In the report a new modified technique of complicated pancreatitis treatment by laparostomy was showed. In included the use of ultrasound dissector for removing necrotic tissues and locating of garamycin sponge which should protect against secondary infection. Between January and May 1998 12 patients were treated using this method. In 8 cases (66%) we obtained rapid improvement of patients' general condition. One patient died and in 6 cases postoperative complications occurred. The use of ultrasound dissector shortens time of necrotic demarcation in pancreas and peripancreatic tissues. Garamycin sponge is effective protection from late infections concerning laparostomic wound. Modified laparostomy decreases about 50% number of secondary re-laparostomies, period of hospitalization and cost of complications treatment in acute pancreatitis in comparison with conventional method. PMID- 10806923 TI - [Registration of hospital infections: effects of 5 years observation of peritonitis]. AB - In connection with a great availability of antibiotics, many negative side effects have been observed. A registration program of hospital infections' occurrence and drug-sensitiveness has been conducted for five years in all wards of Mining Hospital in Sosnowiec. This paper presents our first data and conclusions after examinations of the biological materials coming from patients suffering from different kinds of the peritonitis. PMID- 10806924 TI - [Primary prevention of essential hypertension among doctors and medical students]. AB - Doctors' and medical students' attitude to the primary prevention of hypertension is important, as this community is responsible for medical education. The level of acceptance for some antihypertensive strategies was assessed in 180 doctors and 300 medical students (263 women and 217 men). The study consisted of reading and filling the anonymous questionnaires. The necessity of primary prevention of hypertension in absolutely essential in medical education. PMID- 10806925 TI - [Inflammatory abdominal aortic aneurysm]. AB - The reported incidence of inflammatory abdominal aortic aneurysm (IAAA) is from 2% to 14% of patients with abdominal aortic aneurysm and the etiology of this disease is still discussed--according to the literature several pathogenic theories have been proposed. From 1992 to 1997 32 patients with IAAA were operated on. The patients were mostly symptomatic--abdominal pain was present in 68.75% cases, back pain in 31.25%, fever in 12.5% and weight loss in 6.25% of the operated patients. In all the patients ultrasound examination was performed, in 4 patients CT and in 3 cases urography. All the patients were operated on and characteristic signs of inflammatory abdominal aortic aneurysm like: thickened aortic wall, perianeurysmal infiltration or retroperitoneal fibrosis with involvement of retroperitoneal structures were found. In all cases surgery was performed using transperitoneal approach; in three cases intraoperatively contiguous abdominal organs were injured, which was connected with their involvement into periaortic inflammation. In 4 cases clamping of the aorta was done at the level of the diaphragmatic hiatus. 3 patients (9.37%) died (one patient with ruptured abdominal aortic aneurysm). Authors present diagnostic procedures and the differences in the surgical tactic, emphasizing the necessity of the surgical therapy in patients with inflammatory abdominal aortic aneurysm. PMID- 10806926 TI - [Restoration of the left atrial mechanical function after successful electrical cardioversion]. AB - Some papers indicate that restoration of the left atrial mechanical function (LAMF) can be delayed to even 7 days after successful electrical cardioversion. The goals of the paper include the estimation of delay in restoration of LAMF and factors that influence it. 75 patients (53 males and 22 females, av. age 56.5 +/- 10.8) after elective electrical cardioversion underwent daily echocardiography to determine LAMF, taking A/E ratio > 1/3 as a criterion. The patients were divided into 2 subgroups: G1--with restoration of LAMF < 24 h after cardioversion and G2: the remaining ones. The analysed parameters were: left atrium diameter, rheumatic valvular disease, duration of atrial fibrillation, global and segmental contractility disorders. The data were analysed with ANOVA. The restoration of LAMF < 24 h was found in 53 patients; on the 2nd day in 6 patients, on the 3rd 8th day in 14 patients. In 2 patients no LAMF could be found on the 14th day after cardioversion. Failure to restore LAMF on the cardioversion day was related to: left atrium enlargement, rheumatic valvular disease, duration of atrial fibrillation > 14 days and contractility disorders. PMID- 10806927 TI - [The outline of diagnostic procedures in the cases of focal lesions in liver]. AB - The majority of focal lesions in liver is typified by none or scant clinical symptoms and the first diagnosis is usually made during the ultrasonographic examination (USG). The first examinations usually performed in these cases are: the assignment of neoplastic markers in the serum (alpha-fetoprotein--AFP, carcinoembryonal antigen--CEA), imaging methods (USG, USG using Doppler effect, computer tomography--CT) and isotopic methods. The goal of the study is to present the proposal of outline of diagnostic procedures in the cases of focal lesions in liver based on the data from references. As the first step of diagnostic procedures non-invasive screening examinations are recommended (AFP, CEA, USG, USG using Doppler effect, CT of the abdominal cavity). The increase of the concentration of CEA in serum can suggest the metastatic tumour from the large bowel. The increase of the concentration of AFP makes possible to distinguish hepatocellular carcinoma from other malignant lesions. USG is useful to detect focal lesions which diameter is over 1.5 cm. USG has limited value to qualify the nature of the tumour. On the base of echogenicity of examined lesion we are able to make an initial selection to distinguish single from multiple and cystiform from solid lesions. USG using Doppler effect to detect vessel-signal helps to distinguish sound parenchyma from pathologically changed tissues. CT qualifies focus or dissemination of the disease. After screening examinations we use specialistic or invasive methods which are usually performed in the clinical circumstances. CT connected with angiography has a high sensitivity in detecting hepatocellular carcinoma. Isotopic methods help to confirm the depletion of sound parenchyma of the liver (colloidal scintigraphy) as well as sensitive and specific diagnostics of some types of neoplasms (scintigraphy using labelled erythrocytes to detect angioma). Guided fine-needle biopsy under the USG control makes possible to evaluate the nature of the tumour by means of cytological examination. Diagnosis made by means of sonolaparoscopy and nuclear magnetic resonance (NMR) is described in references, but these examinations have not been used in Poland by this time. Methods which were presented make possible to assign the outline of diagnostic procedures. The outline will help to differentiate precisely and quickly the nature and size of focal lesions in liver. PMID- 10806928 TI - [Psychological aspects of decision-making by women about using hormone replacement therapy]. AB - The number of women living to advanced age increases and also escalates an interest in efforts to prevent or reduce morbidity and premature mortality during the postmenopausal period. The use of postmenopausal hormone replacement therapy (HRT) is recommended by many investigators. While the prevalence of hormone therapy use has been studied, there is limited understanding of how women make a decision about whether or not to adopt hormonal therapy and continue its use. The number of women using HRT varies from 9 to 69%, and also the time of using it is different. Many investigators in Western Europe and in USA found that less educated women were less likely to opt for HRT. Professionally active women, women having children formed the group of estrogen users. The doctors advice has an important significance in making decision about using HRT. In our country there is a need for investigation about using HRT. PMID- 10806930 TI - [Lyme disease: a case report]. AB - A male patient aged 38 was complaining from severe musculo-arthral pains and high body temperature of 39 degrees C. He was admitted to the clinical hospital after long and detailed diagnostic procedures--conducted during previous hospitalization, which focused on Plasmocytoma. Serological examinations relating to Lyme disease were undertaken in the clinical hospital and the results appeared positive. The patient received Vibramicin for a prolonged treatment lasting 30 days. Radiological findings showed distraction in both hip joints (especially the right one), with areas of bone erosion, severe osteoporosis and calcifications in periarthral soft tissues. After pharmacological treatment the body temperature normalized and the pain became less severe. The state of the patient's health improved from subjective and objective point of view. PMID- 10806929 TI - [The second reconstructive operation of the esophagus]. AB - What we need in order to obtain good results in oesophageal reconstructive surgery is long enough segment of intestine with sufficient blood supply. Evaluation of the blood flow is very difficult and often only after completion of the mobilization and transposition of the intestine into the neck final evaluation can be made. In the cases with poor blood flow one must resign from the already mobilized intestine. In such situation the second operation with the use of different alimentary tract segment must be performed. In our practice we encountered 7 cases where it was necessary to mobilize the second intestinal segment in order to achieve success in the reconstruction of the oesophagus. In 3 cases during the first operation jejunum was used. In 3 cases the necrosis of mobilized jejunal segment forced us to perform the second operation. In two patients during the second operation right colon segment along with terminal ileum was used. The third was operated with the use of right colon. In 2 others right colon was used in the first operation. One of them was operated for the second time with the use of left colon. The other were operated with the use of ileum. 2 patients with the substitute from right colon along with the terminal ileum when operated for the second time had the jejunum and left colon segment used to bridge the gap. Mortality in our reconstructive operations was 28.57% (2/7). PMID- 10806931 TI - [Omental torsion in pregnant woman]. AB - A case of triple omental torsion in the 17th week of pregnancy is presented. A 29 year old woman, gravida 3, para 2 was admitted to the hospital because of acute right sided, lower abdominal pain. Clinical symptoms suggested acute appendicitis and/or peritonitis. Due to the worsening of abdominal pain and tachycardia the laparatomy was performed. A 15 cm tumor of omentum twisted around its pedicle was found. It must be stressed that the correct diagnosis was made only after laparotomy. The omentum and appendix were excised. Further postoperative course and pregnancy was uneventful and the woman delivered at term. Clinical difficulties with the correct preoperative diagnosis of this rare entity are discussed. PMID- 10806932 TI - Brackets, epitopes and flash memory cards: a futuristic view of clinical orthodontics. AB - Orthodontics continues to be a profession anchored in traditional technology using appliances that cause inflammatory periodontal ligament (PDL) responses. Existing concepts of biological tooth movement based largely on histological tissue observations and the application of physical principles require major reassessment. In the next millennium, the genome revolution and knowledge of protein production and control could lead to the genetic correction of dentofacial anomalies and pain-free, biomolecular methods of malocclusion correction and long-term stability. A fundamental change is likely to be the abolition of bracket systems and their replacement with preprogrammed microchips driven by computers, and the control of PDL blood vessels and cells by pharmacological targeting. Future survival of the profession will depend on a radically different specialist who will be educated with a postgraduate curriculum based on molecular biology and computer engineering. PMID- 10806933 TI - Bone induction using autogenous bone mixed with demineralised bone matrices. AB - The aim of the present study was to examine the healing of endochondral (EC) autogenous bone grafts in the presence of demineralised bone matrix prepared from intramembranous bone (DBMIM), or prepared from endochondral bone (DBMEC) using quantitative analysis. Thirty bone defects were created on the parietal bone of fifteen New Zealand White rabbits. In the experimental groups, five defects were grafted with EC bone, five defects were grafted with EC bone mixed with DBMIM (EC DBMIM) and six defects were grafted with EC bone mixed with DBMEC (EC-DBMEC). In the control groups, ten defects were left empty (passive control) and four defects were grafted with rabbit skin collagen (positive control). They were all sacrificed at day fourteen post grafting, and the defects were prepared for histological analysis. Serial sections were cut across the whole defect. Quantitative analysis was performed on 152 sections of the experimental groups by image analysis. Four hundred and fourteen per cent more new bone was formed in defects grafted with composite EC-DBMIM than those grafted with EC bone alone (p < 0.001). Eighty-five per cent more new bone was formed in defects grafted with composite EC-DBMEC than those grafted with EC bone alone (p < 0.001). No bone was formed in either passive or positive controls. In conclusion, DBM, especially DBMIM, have extremely high osteoinductive properties and greatly enhance the integration of EC bone grafts with defects created in IM bone. PMID- 10806934 TI - Smoking patterns in adolescent orthodontic patients and school children. AB - Smoking is known to be widely practised by young adults and teenagers, a fact that is of immediate importance to the orthodontic profession as it is now well accepted that smokers have poorer periodontal health than non-smokers. The aim of the present study was to determine the level of smoking in teenage orthodontic patients and compare it with that in the general teenage population and, further, to examine how these smoking habits are influenced by the subjects' attitudes. Patients were surveyed as to their smoking habits and attitudes. All were between 10 and 18 years of age and undergoing active fixed-appliance therapy. The same survey was distributed to a local primary (elementary) school and local secondary (high) schools. Of the 729 school respondents, 29 per cent were classified as smokers; of the 333 respondents in the orthodontic group, 11 per cent were classified as smokers. Although the level of smoking amongst orthodontic patients appears to be lower than that for the general adolescent population, significant numbers of patients smoke on a regular basis, and a small proportion of these is under 13 years of age. Because they have regular contact over a number of years with their adolescent patients, orthodontists are in a favourable position to reinforce non-smoking behavior. PMID- 10806935 TI - Longitudinal cephalometric changes in the maxilla, mandible and maxillary mandibular relationship between 10 and 14 years of age. AB - In the present study, longitudinal growth changes in the maxilla, mandible, and maxillary-mandibular relationship occurring between the ages of 10 and 14 years were evaluated in 19 female and 15 male subjects. All subjects had a clinically acceptable occlusion, a normal growth pattern, and none had undergone orthodontic treatment. Serial cephalometric radiographs were taken at the ages of 10, 11, 12, and 14 years. The effects of age and sex on the sagittal growth of the jaws were studied by means of variance analysis. The results show that A-Ptm, B-Ptm, Pg Ptm, A'-Ptm, B'-Ptm, and S-N-Pg measurements were affected by age, and the measurements A-Ptm, A'-Ptm, Wits and ANB by sex. Duncan's multiple range test was applied to those measurements where F values were found to be statistically significant. As a result, it was observed that the greatest growth changes occurred between 12 and 14 years old. PMID- 10806936 TI - The community and orthodontic care. Part II: Community-perceived importance of correcting various dentofacial anomalies. Part III: Community perception of the importance of orthodontic treatment. AB - Part II. A professionally-managed telephone survey was undertaken to assess the community-perceived importance of correcting various dentofacial anomalies. The sample included 505 respondents, aged eighteen and over, from metropolitan and non-metropolitan households across the state of Victoria. The sample distribution had a 95 per cent confidence limit with a 5 per cent margin of error and closely matched the known population distributions for age, sex and geographical location. This article forms part two of a series. It was found that the correction of functional problems such as "difficulty chewing or speaking" was considered to be very important, regardless of age, sex or geographical area. The correction of other factors such as "top teeth which stick out in front", "bottom teeth which stick out in front" or "crooked or crowded front teeth" was also considered to be important. "Spaced front teeth" was the factor considered least important for correction within all groups. It is interesting to note that, for all factors, correction seemed to be considered more important by females and non metropolitan respondents than by males and metropolitan respondents, In contrast to previous studies in which it has been suggested that patients seek treatment mainly for reasons of aesthetics, the results of this study have shown a definite community recognition of the importance of functional problems as well. Part III. A professionally-managed telephone survey was undertaken to assess the community's perceptions of the importance of having "straight teeth and a nice smile", to assess if a Medicare (the Australian government health benefit scheme) rebate should be provided for orthodontic treatment and to assess whether respondents had any private health insurance that would help cover the cost of orthodontic treatment. The sample included 505 respondents, aged eighteen and over,, from metropolitan and non-metropolitan households across the state of Victoria. The sample distribution had a 95 per cent confidence limit with a 5 per cent margin of error and closely matched the known population distributions for age, sex and geographical location. It was found that a very large percentage of respondents considered the need for "straight teeth and a nice smile" to be very important. This finding is supported by the many studies showing the importance of facial attractiveness to the lives of all people, young and old. Only a small percentage of respondents indicated that they had any private dental health insurance that would help cover the cost of orthodontic treatment. Efforts should be made to inform those responsible for the planning of orthodontic services in both the public and private sectors of the importance of the community's perceptions of aesthetics, and the large amount of published work that reinforces the impact of facial attractiveness on people's lives. PMID- 10806937 TI - Orthodontic management of a patient with epidermolysis bullosa. AB - A male patient with a dental malocclusion presented with Epidermolysis Bullosa, a group of genetically determined diseases characterised by abnormal fragility of the skin and mucosa. Described is the management of the patient utilising Begg Light Wire fixed-appliance orthodontic treatment. PMID- 10806938 TI - A comparison of autotransplantation and orthodontics in a case exhibiting two ectopic upper cuspids. AB - Bilateral severely ectopic maxillary cuspids were treated differently on each side. On one side the ectopic cuspid was moved into position orthodontically, and on the other, the ectopic cuspid was positioned by autotransplantation. The orthodontic positioning took a long time and the tooth exhibited root resorption. Treatment time for the autotransplantation positioning was far quicker, and there was no resorption nor were there any other side effects. PMID- 10806939 TI - Begg appliance management of a moderately crowded Angle Class 11 Division 2 malocclusion. AB - A case is presented that shows Begg appliance management of an Angle Class II division 2 malocclusion. The judicious use of anchorage bends and Class II elastics enabled the successful treatment of the poor posterior and anterior relationships, at the same time addressing the chief complaint of crooked front teeth. PMID- 10806940 TI - An extended three-stage treatment to correct a severe skeletal and dental discrepancy. AB - A report of a case of extended treatment of a patient with severe mandibular retrognathism and an Angle Class II division 1 malocclusion. Initial fixed appliance treatment reduced incisor protrusion. A second phase utilising a Teuscher appliance achieved improved facial and dental relations through excellent mandibular growth. A third phase of fixed Edgewise treatment finalised dental relations. PMID- 10806941 TI - Radiation reduction using a modified collimated lateral skull radiograph during orthodontic treatment. AB - The aim of the study was to investigate whether routine 'during-treatment' clinical cephalometric measurements could be obtained from a lateral skull radiograph collimated to show the maxilla and mandible. The sample consisted of 30 lateral skull radiographs, taken during treatment, of patients with upper and lower fixed appliances. These conventional cephalograms (CC) were copied to give a radiograph with modified collimation (MC) showing the maxilla and mandible and the dentition only. Both types of radiographs were digitised and the readings compared to determine whether the same during-treatment cephalometric values were obtained from both types of radiographs. The limits of agreement obtained, when comparing cephalometric values obtained from the CC and MC lateral skull radiographs, were only marginally larger than those for CC alone. Therefore, the use of an MC lateral skull radiograph to show the maxilla and mandible is a viable alternative for cephalometric measurements for patients wearing two arch fixed appliances. PMID- 10806942 TI - Clinical comparison between a resin-reinforced self-cured glass ionomer cement and a composite resin for direct bonding of orthodontic brackets. Part 2: Bonding on dry enamel and on enamel soaked with saliva. AB - The purposes of this investigation were to compare the clinical performance of a resin-reinforced self-cured glass ionomer cement to a standard composite resin in the direct bonding of orthodontic brackets when bonded onto: a) dry teeth and b) teeth soaked with saliva. The two bonding agents were compared using a split mouth design. In that, both systems were used for direct bonding of stainless steel brackets in every patient. Thirty-eight consecutive patients with fixed appliances were followed for a period of 12 months. The patients were randomly divided into two groups: group A (11 patients) and group B (27 patients). In group A, the performance of 220 stainless steel brackets was evaluated: 110 brackets were bonded with GC Fuji Ortho glass ionomer cement (GC Industrial Co., Tokyo, Japan) onto dry teeth, and 110 bonded with System 1+ composite resin (Ormco Corp., Glendora, CA). In group B, the performance of 540 stainless steel brackets was evaluated: 270 brackets were bonded with GC Fuji Ortho onto teeth soaked with saliva, and 270 bonded with System 1+. In group A, GC Fuji Ortho recorded an overall failure rate (34.5%) significantly higher (p < 0.05) than System 1+ (9%) when applied onto completely dry teeth. Conversely, in group B, no statistically significant differences (p > 0.05) between the failure rates of the two bonding agents were found when GC Fuji Ortho was used on teeth soaked with saliva. It was concluded, therefore, that GC Fuji Ortho shows clinically acceptable bond strengths when bonded onto moist teeth, but not when used on dry enamel. Both bonding agents failed mostly at the enamel/adhesive interface, without causing any enamel damage. PMID- 10806943 TI - Reliability of a method for the localization of displaced maxillary canines using a single panoramic radiograph. AB - The present study was initiated to determine the validity and reproducibility of a method previously reported for localization of displaced maxillary canines by panoramic radiographs. Eleven dental specialists (five orthodontists, five surgeons, and one radiologist) and five (final year) dental students were asked to interpret initial panoramic radiographs of 20 patients with 26 unerupted canines. The method contends that, provided that the radiographic image of the unerupted canines superimposes on the root of the lateral incisor at a height coronal to the apical third, a ratio between the widest mesiodistal dimension of the affected canine and the homolateral central incisor [Canine-Incisor Index (CII)] of more than 1.15 indicates palatal displacement. Among the observers, the measurement error was small, insufficient to cause overlap between the ranges of values for the CII of buccal and palatal canines. Inter-observer reproducibility was high. Without exception, all observers correctly diagnosed the location of each of the impacted canines. The CII cut-off point of 1.15, which was arbitrarily chosen in the previous work, was shown to be valid to differentiate buccal from palatal displacement. The present method is rapid, simple, accurate, and easily taught to dental students or dental specialists, with the simplest of initial instruction. While confirmations with other radiographic techniques is strongly advised before a definitive treatment is undertaken, this method has merit in providing more information than has been available from the panoramic radiograph hitherto, to satisfy the needs of an initial orthodontic consultation. PMID- 10806944 TI - Novel esthetic bonded retainers: a blend of art and science. AB - Novel esthetic S2-glass fiber-reinforced composite wires were fabricated in round (0.022 in.) and rectangular (0.021 x 0.028 in.) profiles, with flexural moduli comparable to martensitic nickel titanium and flexural strengths comparable to beta titanium wires. These wires were bonded clinically to the facial surfaces of teeth in three situations in which flexible-bonded retainers are conventionally used: the retention of a closed median diastema, the retention of space closure following premolar extraction, and the retention of a canine that had been severely malpositioned. These bonded esthetic wires were monitored clinically up to 12 months to ascertain patient's acceptance and structural integrity of the retainer. No clinically observable defects were noted during the study, and no patient described any adverse intraoral effects from the retainer's presence. In this report and on the site http://++www.clinorthodres.com/cor99-c-056, the indications for, and the present design philosophy of, flexible-bonded retainers are discussed, along with the wire properties necessary to achieve optimal stability of tooth position. In addition to demonstrating excellent esthetic characteristics, these photo-pultruded composite wires satisfy the flexural requirements necessary to achieve physiologic stability when used for flexible bonded retention. PMID- 10806945 TI - Surgical orthodontic treatment of skeletal Class III malocclusion with anterior disc displacement without reduction (ADNR): a case report. AB - A female patient with skeletal problems and left temporomandibular joint (TMJ) derangement was treated with an occlusal splint, arthroscopic irrigation, and orthodontic surgery. The left side disc was displaced anteriorly without reduction; and mobility of the left condylar head was restricted. With arthroscopic irrigation, the jaw functions were recovered, but the disc position remained the same. After TMJ therapy, orthodontic and orthognatic surgery treatments were performed to correct the dentofacial deformity. Stable facial esthetics and occlusion devoid of temporomandibular joint disorder (TMD) symptoms were obtained and the patient's progress was monitored over a 5-year period. PMID- 10806946 TI - Extraction site evolution in orthodontics: "the great second molar debate". PMID- 10806947 TI - Second molar replacement applied to dentofacial orthopedics and orthodontics much more than a technique. AB - There is seldom a consensus on any dental subject. The literature contains conflicting points of view and results of studies vary considerably. I have endeavored to present my paradigm of second molar replacement in dentofacial orthopedics and orthodontics. I have presented a summation of information known to date regarding this subject. I based this article on my extensive clinical experience and observations over many years. I participate in continuing education, and know many of the traditional and functional post-doctoral educators. I read the literature. For your benefit, I have presented this information and shared a paradigm that has positively changed my life, and perhaps will change the way you practice. Second molar replacement technique has revolutionized orthodontic treatment. The technique offers distinct benefits considering the patient's face, the jaw joints, and the teeth. The reader is strongly encouraged to read bibliography references for a mature understanding of second molar replacement in modern orthodontics and TMD therapy. When extraction of teeth is needed for orthodontic purposes Broadbent considers second molar replacement, second bicuspid removal, first bicuspid removal, lower incisor removal, and then other extraction possibilities to resolve the malocclusion. When extraction guidelines are followed and treatment mechanics are properly designed the extraction of second molars have predicable good results in a high percentage of cases. However, second molar removal may be contraindicated for numerous reasons. Diagnosis and case selection is paramount! The doctor must diagnosis and determine if and when second molar replacement would be in each patient's best interest. The bottom line for each individual patient is the best possible facial esthetics, functional occlusion, and health of jaw joints. PMID- 10806948 TI - The greatest breakthrough in orthodontics of the century. PMID- 10806949 TI - An update on the indirect-bonding technique (non-custom base) PMID- 10806950 TI - TMD: it's our responsibility. AB - Temporomandibular disorder (TMD) is a reality. Surveys place the incidence at about 20-50%. For decades debate has raged as to whether this is a medical or dental problem. Scientific study of anatomy, biochemistry, physiology, and objective analysis with kinesiology, electromyography and sonography, along with case history all point to the same thing; occlusion affects the joint and the muscles. This is our profession's "crown jewel". We diagnose, construct and modify our patient's occlusion. It is about time we all agree, understand, take responsibility and start cooperating in preventing and treating this common malady that seriously affects the quality of life of many. PMID- 10806951 TI - The controlled arch system: a new method of straightwire treatment. AB - The "Controlled Arch System", coupled with a proper diagnosis and treatment plan, should produce excellent occlusion and esthetics for your patients from their Mixed Dentition growth period onward. The authors have offered a method of Phase I treatment for children of Mixed Dentition age that can be outlined as follows: 1. Fit maxillary and mandibular Functional Orthopedic appliances to produce whatever transverse expansion of the arches is needed, then distalize the maxillary 6 year molars to a super Class I relation, according the measurements assessed by the Sim Model Analysis. 2. On removal of the Functional Orthopedic appliances, upper and lower Fixed-Removable Lingual Arches are fitted to stabilize teeth and bone. 3. As needed, fit maxillary and mandibular 2 x 4 or 2 x 6 fixed Straightwire appliances with Nickel Titanium wires, utilizing pinched molar hook/stops to establish molar anchorage to level, align and rotate permanent incisors (and lower permanent canines, if erupted). 4. When alignment of the permanent incisors is completed, the FRLAs are left in place as "insurance" appliances to insure that no loss of arch width or arch length occurs. The FRLAs are left in place for up to two years as retainers. 5. Be sure to inform parents and patients that Phase II comprehensive fixed Straightwire treatment is almost certain to be needed during adolescence when the 28 permanent teeth have erupted. 6. Use of this "Controlled Arch System" not only simplifies and shortens the duration of orthodontic treatment, but also can dramatically lower the percentage of extraction cases in an orthodontic practice. PMID- 10806952 TI - TMD: it's our responsibility! Part Two. AB - Dentists modify and construct occlusions. This occurs on a daily basis in the disciplines of restorative, fixed and removable prosthetics and orthodontics. These procedures influence the TMJ, the muscles of mastication, the supporting structure of the teeth and the teeth as well. Dentists have an opportunity to not only objectively analyze how they all interrelate, but to create a physiological harmonious relationship. This will reduce a traumatic occlusion to one that is within the histological adaptive range for the tissue to accommodate. This knowledge can be applied to not only help patientsi pain and dysfunction, but to ensure confidence that dentists are not going to be responsible for iatrogenic results. The future of dentistry is not only being able to effectively deal with TMD but to prevent the problems. Management of mandibular whiplash and migraine headaches are also becoming our responsibility. Dentists are the specialists of occlusion. With the technology available to render this quality of care, they must fulfill their obligation to the public, take responsibility and continue to increase their capability. This approach to dentistry provides a new "window of opportunity" as its many applications have yet to be explored. PMID- 10806953 TI - Marketing for orthodontics: guidelines for the generalist and pediatric dentist. PMID- 10806954 TI - Recognizing child abuse and neglect: a survey of Kansas dental offices. PMID- 10806955 TI - Tips for managing patients with Alzheimer's disease. PMID- 10806956 TI - Enterovirus infections with special reference to enterovirus 71. AB - The enteroviruses comprise a large group of immunologically distinct serotypes of viruses belonging to the family of Picornaviridae. Many enteroviruses cause diseases in human, but the infections are generally mild as asymptomatic, therefore, enteroviruses are considered to be unimportant as human pathogens. However, enteroviruses may also result in serious or even fatal disease (as shown in the enterovirus 71 (EV71) epidemic in Taiwan in 1998). There are three types of polioviruses, Coxsackievirus group A and group B viruses, and echoviruses group. All together a total of 67 types are available. Starting from enterovirus type 68 to 71, they are named as enterovirus types. Enterovirus type 72 is hepatitis A virus. Paralytic disease of poliomyelitis was recorded in ancient time but characterization of poliovirus was not reported until the turn of the 19th century that poliomyelitis was a viral disease. The major breakthrough for diagnosing and controlling of poliomyelitis was the discovery that poliovirus can be propagated in human embryonic tissues in cultures. As soon as cultures of human and monkey cells began to use for isolating polioviruses in stool specimen of patients, more unknown viruses were isolated which unlike polioviruses nor Coxsackie viruses; they were called "orphan" viruses or human enteric viruses, name later simplified to "echoviruses". Morphologically all enteroviruses are alike. They are small, ether insensitive viruses with an RNA genome. Their nucleic acid is single stranded, and the nucleocapsid has a cubic (icosahedral) symmetry, and is naked. The host ranges of enteroviruses vary greatly from one type to the next and even among strains of the same type. Polioviruses have a very restricted host range among laboratory animals. Virus isolation is the best method for diagnosis of enterovirus infection, but infection in the central nervous system (CNS) may be detected by polymerase chain reaction (PCR). Currently final identification and serotyping of enteroviruses are by indirect immunofluorescent tests using monoclonal antibody or by neutralization test using antiserum pools described by Lim and Benyesh-Melnick. The incidence and prevalence of diseases associated with the enterovirus infections are varied. The circulation of enteroviruses recently in Tainan and the epidemic of EV71 in Taiwan in 1998 are described in this review. Although poliovirus infection may be eradicated from the world due to the efficient vaccination program, there is no specific antiviral agents for either treatment or prevention for other enterovirus infections. In 1991, a new antiviral "pleconaril" which is a novel orally bioavailable and systematically acting small molecule inhibitor for picornaviruses. "Pleconaril" is currently in clinical trials for treatment of enterovirus meningitis and respiratory infections. PMID- 10806957 TI - Identification of virus-specific vesicles in Giardiavirus-infected Giardia lamblia. AB - Giardiavirus (GLV), which infects the parasitic protozoan Giardia lamblia, is a nonsegmented double-stranded (ds) ribonucleic acid (RNA) virus. We previously purified two distinct types of related GLV from infected G. lamblia, and showed differential export of one of the viruses from infected cells. In the present study, fractionation of cell lysate was performed, revealing the presence of viruses in the membranous fraction. Distribution of viral antigens in the infected cells was examined by immunocytochemistry. The signal was enriched in certain regions of the cytoplasm, suggesting that a portion of GLV is confined to certain cellular compartments. A significantly reduced signal was also detected in the nuclei. We directly observed the viruses in the infected cells by electron microscopy. Consistent with previous observations, virus-like particles were clearly observed in some membranous vesicles in the cytoplasm at 48 h postinfection, and virus-like particles were again seen in the cytoplasm and then in the nuclei toward the late phase of virus infection. The virus-associated vesicles and some electron-dense nuclear structures were only observed in virus infected cells, suggesting that virus infection may induce ultrastructural alteration of G. lamblia. PMID- 10806958 TI - TT virus infection among hemodialysis patients at a medical center in Taiwan. AB - Although the association between TT virus (TTV) infection and hepatitis is controversial, the high prevalence of TTV infection in healthy blood donors and even higher rate among frequently transfused patients poses a potential threat to public health and clinical care. In addition, there is a lack of data concerning the prevalence and mode of transmission of TTV infection in different subpopulations in Taiwan. In the present study, we investigated the prevalence of TTV infection in 111 uremic patients receiving regular hemodialysis in a single hospital in Taiwan. Blood samples were collected and analyzed using a seminested polymerase chain reaction (PCR) designed to amplify a 271 base-pair DNA fragment. The results show that the overall TTV positive rate in uremic patients in our hospital was 61% (68/111), which was much higher than the reported TTV prevalence rate among the normal population (ranging from 1%-12%). The results of analysis of the demographic and clinical characteristics of the patients indicate that blood transfusion may play an important role in TTV transmission (p < 0.05). In addition, the hepatitis B positive rate was significantly lower in TTV positive patients. However, liver function tests were not significantly different between TTV positive and TTV negative patients. The results of the present study suggest that blood transfusion plays an important role in TTV transmission in uremic patients. PMID- 10806959 TI - Profiles of inflammatory cytokines in bronchoalveolar lavage fluid from premature infants with respiratory distress disease. AB - In view of cytokine's effects in promoting or inhibiting inflammation, the objective of this study was to explore the characteristics of the proinflammatory cytokine, interleukin-8 (IL-8), and the inhibitory cytokine, interleukin-10 (IL 10), in the bronchoalveolar lavage (BAL) fluid of premature infants suffering from respiratory distress disease. Eighteen premature neonates with respiratory distress disease with gestational age (GA) ranging from 24 to 37 weeks were recruited for study. BAL fluids were collected following endotracheal intubation during an episode of hypoxemia or dyspnea. A series of BAL samples were obtained on day 1, 2, 4 and 7 after intubation for measuring IL-8 and IL-10 levels. The results indicate that premature infants with GA ranging from 24 to 32 weeks had a higher level of IL-8 (p = 0.029), but not level of IL-10 (p = 0.109), in the BAL obtained during the first intubation compared to premature infants with GA ranging from 33 to 37 weeks. The administration of exogenous surfactant did not influence the profiles of IL-8 and IL-10, as compared to those in-patients without treatment. Levels of IL-8 were correlated with IL-10 levels (r = 0.613, p = 0.007) in BAL fluid samples obtained on the day of intubation. The level of IL 8, but not IL-10, was significantly correlated with the duration of intubation. IL-8 and IL-10 levels in BAL fluid samples collected on the day of intubation were correlated with the development of chronic lung disease (CLD). The results suggest that extreme prematurity tends to have increased IL-8 and IL-10 levels in BAL fluid compared to premature infants with older GA, and that these increased levels are associated with the development of CLD. PMID- 10806960 TI - Acute effect of glucocorticoid treatment on serum osteocalcin levels in asthmatic children. AB - The serum levels of osteocalcin (OC), a sensitive and specific biochemical marker of bone formation, were measured in 36 asthmatic children before and after intravenous administration of glucocorticoid (GC), methylprednisolone (1 mg/kg/6 h). A statistically significant (p < 0.005) decrease in OC level (13.5-2.7 microg/L) after the completion of 1-day treatment. In patients who received 2-day treatment, the OC levels further declined and reached about 20% of the pretreatment levels. Blood samples collected from patients who received 3-day treatment showed serum OC had declined to an even lower level after the completion of therapy (11.5-1.8 microg/L). An inverse relationship was found between the OC concentration and the duration of GC therapy. GC therapy induced a minor, significant decrease of serum alkaline phosphatase level but did not influence serum calcium level. A tendency toward decrease of serum phosphate level was also noted during GC treatment. An acute effect of GC therapy on the suppression of osteoblasts was biochemically revealed by the finding of reduced serum OC levels. The results suggest that early change in serum OC may be a useful indicator for patients at high risk of bone loss. PMID- 10806961 TI - Application of pulsed-field gel electrophoresis to the investigation of a nosocomial outbreak of Vibrio parahaemolyticus. AB - The method of pulsed-field gel electrophoresis (PFGE) for Vibrio parahaemolyticus was first published in 1996. Since then, its application has been seldom reported in the literature. A food poisoning outbreak due to V. parahaemolyticus occurred in five wards of a hospital was investigated using this method. Twenty-five patients were involved and all of them had eaten food supplied by the hospital. Of the 15 cases whose stools were available for culture, only four cultures yielded V. parahaemolyticus. All four isolates were serotype K6 and were indistinguishable or closely related to each other based on PFGE patterns. Two isolates from food were recovered and they presented different characterizations from the patient isolates in both serotype and PFGE pattern. Successful typing by PFGE to identify the outbreak strain and differentiate V. parahaemolyticus strains between patient and food isolates in this study suggests the usefulness of PFGE for V. parahaemolyticus, the leading cause of food poisoning in Taiwan. PMID- 10806962 TI - Chlamydia pneumoniae infection in community-acquired pneumonia in Taiwan. AB - Chlamydia pneumoniae is a common cause of pneumonia worldwide. This study examined the role of C. pneumoniae in 229 patients with community-acquired pneumonia. The ages of the patients ranged from 2 to 95 years. Sera were assayed for IgM and IgG antibodies with microimmunofluorescence (MIF). An IgM titer equal to or greater than 1:20 and IgG titer equal to or greater than 1:64 were considered positive. The prevalence of positive IgG among all of the patients was 75.1% (172/229). The seroprevalence was 81.8% (9/11) for patients in the 11- to 20-year-old group, 63.6% (14/22) in the 31- to 40-year-old group and 88.1% (52/59) in the 71- to 80-year-old group. All patients had a negative result for IgM antibody. Twenty-five of the patients had an IgG titer equal to or greater than 1:512, indicating the presence of current infection or reinfection. Age older than 60 years (vs. < or = 60 years) was a risk factor for C. pneumoniae seropositivity in patients with community-acquired pneumonia (p < 0.001). Males had a significantly higher seroprevalence of C. pneumoniae infection (p = 0.1010). Patients older than 60 years were more likely to have C. pneumoniae infection (p = 0.1107). In this series, C. pneumoniae infection accounted for 10.9% (25/229) of community-acquired pneumonia. The most common clinical manifestations included fever (92%), productive cough (52%), white blood cell more than 10,000/mm3 (56%), and bilateral pulmonary infiltrate (60%). PMID- 10806963 TI - An outbreak of Salmonella infection after a Chinese year-end party in central Taiwan. AB - An outbreak of food poisoning caused by Salmonella O7 serogroup C1 and O8 serogroup C2, occurred in Taichung City after a Chinese year-end buffet party with 127 attendees including employees, relatives and guests of the Psychiatry Department of Changhua Christian Hospital (CCH). Among the 114 attendees interviewed, 96 (84.2%) reported developing symptoms within 120 h after the dinner on February 4, 1999. The time of onset ranged from 2 h to 101 h after the dinner with an average of 20 +/- 16 h. The median and mode incubation periods were 17 h and 16 h, respectively. Salmonella C1 and C2 serogroups were isolated from the stool samples of 45 attendees. Based on the results of interview questionnaire, the most likely contaminated food was eel kabayaki (OR = 4.8, 95% CI:1.6-14.9, p < 0.01) followed by baked mussels (OR = 4.04, 95% CI:1.3-12.1, p = 0.01). However, this result could not be confirmed by food sample investigation due to the lack of leftover food. Possible techniques for the prevention of food borne disease transmission, enhancement of communication about foodborne disease outbreaks within the health reporting system, and the reduction of response time during an outbreak of infection are required. PMID- 10806964 TI - Ophthalmic findings in patients with acquired immunodeficiency syndrome. AB - Ocular manifestations have been reported in up to 60% of individuals infected with human immunodeficiency virus (HIV) in the United States, and it is becoming increasing apparent that these ocular manifestations almost invariably reflect extent of progression of the disease. The prevalence of ocular abnormalities among acquired immunodeficiency syndrome (AIDS) patients in Taiwan has not been reported. In the present study, we examined and followed up the ophthalmic conditions of a total of 274 HIV-infected patients during the period from March 1993 to May 1999. The results show that cotton-wool spots was the most common ocular finding in this series of patients with AIDS, occurring in 22 (32.8%) of 67 AIDS patients. Cytomegalovirus (CMV) retinitis was the most commonly seen opportunistic ocular infection, occurring in 14 (20.8%) of 67 AIDS patients. These findings suggest that AIDS patients should be closely followed for signs of opportunistic ocular disease which may initially be asymptomatic. Close co operation between the ophthalmologist and the internist is essential to ensure timely therapeutic intervention, which can decrease the risk of further complications including visual impairment and blindness. PMID- 10806965 TI - The assessment of anaerobic blood culture in children. AB - Over the past 20 years, there has been a decline in the percentage of positive blood cultures yielding anaerobic organisms. Due to the limited blood volume drawn from pediatric patients, we have assessed the value of routine anaerobic blood cultures in children. From January 1994 to December 1998, 9886 paired aerobic and anaerobic blood cultures were analyzed in the pediatric microbiology laboratory at the Taipei Veterans General Hospital. Six hundred and eighteen (6.25%) isolates were considered to be clinically important microorganisms. Staphylococci, streptococci, aerobic gram-negative bacilli, and yeasts showed more significant growth within the aerobic culture than that within the anaerobic culture. Significantly more aerobic gram-positive cocci, aerobic gram-negative bacilli, and yeasts were detected at least 1 day earlier by using the aerobic culture. Three patients with documented anaerobic bacteremia had obvious symptoms related to anaerobic infections. Our study concludes that routine use of anaerobic blood culture in pediatric patients is not necessary. Anaerobic blood cultures should be reserved for patients with diseases like intra-abdominal or oral infections, neutropenic patients on steroid therapy, pressure sores, cellulitis, and human bite wounds. PMID- 10806966 TI - Lichen sclerosus et atrophicus, bullous morphea, and systemic lupus erythematosus: a case report. AB - Lichen sclerosus et atrophicus (LSA) rarely coexists with morphea, especially when bullae occur in lesions of morphea. Here we report the case of a 15-year-old girl with this condition, who also fulfilled four out of 11 diagnostic criteria for systemic lupus erythematosus (SLE). Tissue biopsy of different skin lesions showed LSA in the regions of bullous morphea, that has rarely been reported in the literature. PMID- 10806967 TI - Community-acquired methicillin-resistant Staphylococcus aureus endocarditis with septic embolism of popliteal artery: a case report. AB - A 20-year-old man presented with a 14-day course of fever. Physical examination showed petechiae of the conjunctivae, Janeway lesions on both hands, a grade III/VI systolic murmur over the apex, pulseless dorsal pedal artery and posterior tibial artery of the right leg, and a pale right foot. Femoral arteriogram of the right leg revealed total occlusion of the popliteal artery with collateral circulation of the posterior tibial artery. Transthoracic echocardiogram showed trace mitral regurgitation. Embolectomy of the right popliteal artery was done, and penicillin and gentamicin treatment was administered. However, postoperative fever developed intermittently. Transesophageal echocardiogram disclosed vegetation over the anterior leaflet of the mitral valve. Methicillin-resistant Staphylococcus aureus (MRSA) was isolated from all three cultures of blood drawn at admission and from the septic embolus during operation. He had neither evidence of underlying heart disease, nor history of intravenous drug abuse or hospitalization. Exploratory cardiotomy with removal of vegetation on the mitral valve was performed followed by a 4-week treatment with intravenous vancomycin. After discharge, he was well at 2-year follow-up. PMID- 10806968 TI - Strain FV-21 of simian foamy virus type 1 was cloned and sequenced after isolation from the Taiwan monkey Macaca cyclopsis. PMID- 10806969 TI - Guide to Community Preventive Services: harnessing the science. PMID- 10806971 TI - The Guide to Community Preventive Services: managed care/clinical practice commentary. PMID- 10806970 TI - A health department perspective on the Guide to Community Preventive Services. PMID- 10806972 TI - Commentary on the emerging Guide to Community Preventive Services from a health promotion perspective. PMID- 10806973 TI - Guide to Community Preventive Services: a commentary. PMID- 10806974 TI - The Guide to Community Preventive Services will be influential in academic health centers: education, research and links with practice. PMID- 10806975 TI - Methods for reviewing economic evaluations of community preventive services: a cart without a horse? PMID- 10806976 TI - Developing the Guide to Community Preventive Services--overview and rationale. The Task Force on Community Preventive Services. AB - When the GUIDE TO COMMUNITY PREVENTIVE SERVICES: Systematic Reviews and Evidence Based Recommendations (the Guide) is published in 2001, it will represent a significant national effort in encouraging evidence-based public health practice in defined populations (e.g., communities or members of specific managed care plans). The Guide will make recommendations regarding public health interventions to reduce illness, disability, premature death, and environmental hazards that impair community health and quality of life. The Guide is being developed under the guidance of the Task Force on Community Preventive Services (the Task Force) a 15-member, nonfederal, independent panel of experts. Subject matter experts, methodologists, and scientific staff are supporting the Task Force in using explicit rules to conduct systematic literature reviews of evidence of effectiveness, economic efficiency, and feasibility on which to base recommendations for community action. Contributors to the Guide are building on the experience of others to confront methodologic challenges unique to the assessment of complex multicomponent intervention studies with nonexperimental or nonrandomized designs and diverse measures of outcome and effectiveness. Persons who plan, fund, and implement population-based services and policies to improve health at the state and local levels are invited to scrutinize the work in progress and to communicate with contributors. When the Guide is complete, readers are encouraged to consider critically the value and relevance of its contents, the implementation of interventions the Task Force recommends, the abandonment of interventions the Task Force does not recommend, and the need for rigorous evaluation of the benefits and harms of promising interventions of unknown effectiveness. PMID- 10806977 TI - Scope and organization of the Guide to Community Preventive Services. The Task Force on Community Preventive Services. AB - BACKGROUND: The diverse nature of the target audience (i.e., public health decision-makers) for the Guide to Community Preventive Services: Systematic Reviews and Evidence-Based Recommendations (the Guide) dictates that it must be broad in scope. In addition, for the Guide to be most useful for its target audience, its organization and format must be carefully considered. DETERMINING THE SCOPE OF THE GUIDE: Healthy People objectives and actual causes of death were used to determine the contents of the Guide. A priority setting exercise resulted in the selection of 15 topics for systematic reviews using the following criteria: burden of the problem, preventability, relationship to other public health initiatives, usefulness of the package of topics selected and level of current research and intervention activity in public and private sectors. Interventions within each topic target state and local levels and include population-based strategies, individual strategies in other than clinical settings and group strategies. ORGANIZATION OF THE GUIDE: The Guide is organized into: Introduction, Reviews and Recommendations (three sections: Changing Risk Behaviors, Reducing Diseases, Injuries, or Impairments, and Addressing Environmental and Ecosystem Challenges), Appendixes, and Indexes. DISCUSSION: The scope and organization of the Guide were determined using relevant public health criteria and expert opinion to provide a useful and accessible document to a broad target audience. While the final contents of the Guide may change during development, the working table of contents described in this paper provides a framework for development of the Guide and conveys its scope and intention. PMID- 10806978 TI - Developing an evidence-based Guide to Community Preventive Services--methods. The Task Force on Community Preventive Services. AB - Systematic reviews and evidence-based recommendations are increasingly important for decision making in health and medicine. Over the past 20 years, information on the science of synthesizing research results has exploded. However, some approaches to systematic reviews of the effectiveness of clinical preventive services and medical care may be less appropriate for evaluating population-based interventions. Furthermore, methods for linking evidence to recommendations are less well developed than methods for synthesizing evidence. The Guide to Community Preventive Services: Systematic Reviews and Evidence-Based Recommendations (the Guide) will evaluate and make recommendations on population based and public health interventions. This paper provides an overview of the Guide's process to systematically review evidence and translate that evidence into recommendations. The Guide reviews evidence on effectiveness, the applicability of effectiveness data, (i.e., the extent to which available effectiveness data is thought to apply to additional populations and settings), the intervention's other effects (i.e., important side effects), economic impact, and barriers to implementation of interventions. The steps for obtaining and evaluating evidence into recommendations involve: (1) forming multidisciplinary chapter development teams, (2) developing a conceptual approach to organizing, grouping, selecting and evaluating the interventions in each chapter; (3) selecting interventions to be evaluated; (4) searching for and retrieving evidence; (5) assessing the quality of and summarizing the body of evidence of effectiveness; (6) translating the body of evidence of effectiveness into recommendations; (7) considering information on evidence other than effectiveness; and (8) identifying and summarizing research gaps. Systematic reviews of and evidence-based recommendations for population-health interventions are challenging and methods will continue to evolve. However, using an evidence based approach to identify and recommend effective interventions directed at specific public health goals may reduce errors in how information is collected and interpreted, identify important gaps in current knowledge thus guiding further research, and enhance the Guide users' ability to assess whether recommendations are valid and prudent from their own perspectives. Over time, all of these advantages could help to increase agreement regarding appropriate community health strategies and help to increase their implementation. PMID- 10806979 TI - Data collection instrument and procedure for systematic reviews in the Guide to Community Preventive Services. Task Force on Community Preventive Services. AB - INTRODUCTION: A standardized abstraction form and procedure was developed to provide consistency, reduce bias, and improve validity and reliability in the Guide to Community Preventive Services: Systematic Reviews and Evidence-Based Recommendations (the Guide). DATA COLLECTION INSTRUMENT: The content of the abstraction form was based on methodologies used in other systematic reviews; reporting standards established by major health and social science journals; the evaluation, statistical and meta-analytic literature; expert opinion and review; and pilot-testing. The form is used to classify and describe key characteristics of the intervention and evaluation (26 questions) and assess the quality of the study's execution (23 questions). Study procedures and results are collected and specific threats to the validity of the study are assessed across six categories (intervention and study descriptions, sampling, measurement, analysis, interpretation of results and other execution issues). DATA COLLECTION PROCEDURES: Each study is abstracted by two independent reviewers and reconciled by the chapter development team. Reviewers are trained and provided with feedback. DISCUSSION: What to abstract and how to summarize the data are discretionary choices that influence conclusions drawn on the quality of execution of the study and its effectiveness. The form balances flexibility for the evaluation of papers with different study designs and intervention types with the need to ask specific questions to maximize validity and reliability. It provides a structured format that researchers and others can use to review the content and quality of papers, conduct systematic reviews, or develop manuscripts. A systematic approach to developing and evaluating manuscripts will help to promote overall improvement of the scientific literature. PMID- 10806980 TI - Methods for systematic reviews of economic evaluations for the Guide to Community Preventive Services. Task Force on Community Preventive Services. AB - OBJECTIVES: This paper describes the methods used in the Guide to Community Preventive Services: Systematic Reviews and Evidence-Based Recommendations (the Guide) for conducting systematic reviews of economic evaluations across community health-promotion and disease-prevention interventions. The lack of standardized methods to improve the comparability of results from economic evaluations has hampered the use of data on costs and financial benefits in evidence-based reviews of effectiveness. The methods and instruments developed for the Guide provide an explicit and systematic approach for abstracting economic evaluation data and increase the usefulness of economic information for policy making in health care and public health. METHODS: The following steps were taken for systematic reviews of economic evaluations: (1) systematic searches were conducted; (2) studies using economic analytic methods, such as cost analysis or cost-effectiveness, cost-benefit or cost-utility analysis, were selected according to explicit inclusion criteria; (3) economic data were abstracted and adjusted using a standardized abstraction form; and (4) adjusted summary measures were listed in summary tables. RESULTS: These methods were used in a review of 10 interventions designed to improve vaccination coverage in children, adolescents and adults. Ten average costs and 14 cost-effectiveness ratios were abstracted or calculated from data reported in 24 studies and expressed in 1997 USD. The types of costs included in the analysis and intervention definitions varied extensively. Gaps in data were found for many interventions. PMID- 10806981 TI - Recommendations regarding interventions to improve vaccination coverage in children, adolescents, and adults. Task Force on Community Preventive Services. PMID- 10806982 TI - Reviews of evidence regarding interventions to improve vaccination coverage in children, adolescents, and adults. The Task Force on Community Preventive Services. AB - This paper presents the results of systematic reviews of the effectiveness, applicability, other effects, economic impact, and barriers to use of selected population-based interventions intended to improve vaccination coverage. The related systematic reviews are linked by a common conceptual approach. These reviews form the basis for recommendations by the Task Force on Community Preventive Services (the Task Force) regarding the use of these selected interventions. The Task Force recommendations are presented on pp. 92-96 of this issue. PMID- 10806983 TI - Impedance to gas transfer across the alveolar-capillary membrane in chronic cardiac failure. AB - One of the consequences of an elevation of the pulmonary capillary pressure in chronic heart failure is the occurrence of structural and functional changes at the level of the alveolar-capillary interface. These changes are called "stress failure" of the membrane, and consist of thickening of the interstitium, increase in capillary permeability to water and ions, and disruption of local regulatory mechanisms for gas exchange. Functional correlates are an augmented impedance to gas transfer (DL), a reduction in the alveolar-capillary membrane conduction (DM) and an increase in the volume of the pulmonary capillary blood (VC). DM and VC are the two subcomponents of DL. DM has been identified as the strongest respiratory predictor of oxygen uptake at peak exercise in patients with chronic heart failure, suggesting that impeded lung diffusion may significantly contribute to exercise limitation and ventilatory abnormalities. The evidence relating to the pathophysiological and clinical significance of the impairment in lung diffusion capacity in patients with chronic heart failure, as well as the response to treatment are the main subjects of this review. PMID- 10806984 TI - Transmural heterogeneity of myocardial contraction and ischemia. Diagnosis and clinical implications. AB - Myocardial contraction behaves heterogeneously, being greater in subendocardial than in subepicardial layers. Similarly, during acute myocardial ischemia or infarction, the subendocardium is the first myocardial layer to suffer. Conventional two-dimensional echocardiography cannot distinguish the transmural extension of myocardial ischemia or infarction, showing akinesia also when only the subendocardium is affected. Novel ultrasonographic techniques (like tissue characterization with integrated backscatter or Doppler tissue imaging) and nuclear magnetic resonance tagging can investigate myocardial contraction in different transmural layers and distinguish subendocardial from transmural ischemia or infarction. With the advent of thrombolysis and primary angioplasty in the acute phase of myocardial infarction a correct diagnosis of the extension of myocardial necrosis cannot ignore its transmural wavefront development. The salvage of the subepicardial layer does not give direct information on overall myocardial thickening but is one of the major determinants of overall left ventricular dysfunction and size. Although it is still necessary to investigate this phenomenon, new ultrasonographic techniques give us important information and more opportunities to appropriate diagnosis and future treatment of cardiac patients. PMID- 10806985 TI - Vascular and haemostatic gene polymorphisms associated with non-fatal myocardial infarction: a critical review. AB - The importance of genetics to the pathogenesis of myocardial infarction is suggested by the frequent familial clustering of premature disease. Yet, studies associating myocardial infarction with gene polymorphisms of vascular proteins (angiotensinogen, angiotensin converting enzyme, angiotensin II type 1 receptor, endothelial nitric oxide synthase) and haemostatic factors (fibrinogen, coagulation factors II, V, VII and XIII, plasminogen activator inhibitor-1, tissue-type plasminogen activator, platelet glycoproteins IIb/IIIa, Ia/IIa and Ib IX-V, or methylenetetrahydrofolate reductase) have revealed conflicting results. This is hardly surprising, given: 1) the multigenic nature of myocardial infarction, whereby single polymorphisms are bound to play at best only a limited role in the global risk of disease; 2) the multiple pathogenetic mechanisms of infarction (e.g., atheromatous obstruction, plaque rupture, thrombosis, vasospasm), each of which is likely influenced by a number of genes and by several environmental factors. The simultaneous investigation of a set of polymorphisms--and of their interactions with environmental factors--in extremely homogeneous sets of patients should offer a better understanding of the contribution of specific genes to the risk of myocardial infarction. PMID- 10806986 TI - Pulmonary venous flow and mitral inflow velocity pattern in uncomplicated obesity: evidence for late diastolic dysfunction. AB - BACKGROUND: Active left ventricular relaxation, assessed by Doppler isovolumic relaxation time, is impaired in obesity. There is little information on left ventricular passive properties during filling. METHODS: To evaluate left ventricular late diastolic stiffness in obesity, Doppler echocardiographic interrogation of mitral inflow tract and pulmonary vein flow velocities were obtained from 47 normotensive, young obese subjects (11 males, 36 females) and 43 normotensive, young normal-weight volunteers (13 males, 30 females) of comparable age. RESULTS: After controlling the effect of blood pressure and left ventricular mass, isovolumic relaxation time was prolonged in obese subjects (p < 0.0001 vs normal-weight controls). No difference was found in transmitral peak early and late flow velocities. Obese subjects exhibited prolonged pulmonary vein reverse flow velocity during atrial contraction (p < 0.004), and a higher difference or ratio between duration of pulmonary reverse flow and duration of transmitral forward late flow (6 +/- 31 vs -20 +/- 39 ms or 1.06 +/- 0.3 vs 0.84 +/- 0.3, p < 0.002 and p < 0.001, respectively). These differences were also confirmed after controlling blood pressure and left ventricular mass. Non-invasively estimated left ventricular end-diastolic pressure was higher in obese subjects than in controls (p < 0.002). At multivariate analysis a higher body mass index was the sole predictor of prolonged difference between duration of pulmonary reverse flow and duration of transmitral forward late flow (beta = 0.38, p < 0.001). CONCLUSIONS: Obesity is associated with prolonged left ventricular active relaxation and abnormalities of filling pressure not detectable by the sole mitral inflow velocity pattern. These latter abnormalities are consistent with the presence of early increased left ventricular passive stiffness. PMID- 10806987 TI - Progressive decrease of outflow gradient and septum thickness after percutaneous alcoholization of the interventricular septum in hypertrophic obstructive cardiomyopathy. AB - BACKGROUND: The aim of this study was to evaluate the immediate- and mid-term results of percutaneous transluminal septal myocardial ablation (PTSMA) of the interventricular septum performed in 15 consecutive patients with hypertrophic obstructive cardiomyopathy between 1996 and 1999. METHODS: Prior to intervention, all patients (7 males, 8 females, mean age 62 +/- 11 years) complained of severe dyspnea (NYHA functional class III-IV) despite medical treatment with beta blockers and/or verapamil. Family history of hypertrophic cardiomyopathy was present in 2 cases. Dehydrated alcohol (4.8 +/- 1.5 ml/pt) was selectively infused into the first septal perforator artery through over-the-wire balloon catheters. In 5 patients a second or a third septal branch was treated because the intraventricular gradient persisted above 50 mmHg after the initial alcohol infusion. RESULTS: Alcohol infusion induced an average peak creatine phosphokinase level of 1,524 +/- 427 IU/l. No iterating ventricular arrhythmias occurred during the procedure or in the 2-3 days of continuous ECG monitoring after the procedure. Two patients (13%) developed a complete atrioventricular block after the procedure, requiring permanent double-chamber pacing. Electrocardiographic changes included a > 2 mm ST segment elevation and transient right bundle branch block or left anterior/left posterior hemiblock in all patients. Peak basal intraventricular gradient decreased from 80 +/- 27 to 24 +/- 27 mmHg (p < 0.01) during cardiac catheterization and from 81 +/- 27 to 35 +/- 25 mmHg (p < 0.01) at the echocardiographic control performed during the hospital stay. At follow-up (mean 5.1 +/- 3.6 months), all patients were in NYHA functional class I or II. Repeat echocardiography showed a further significant decrease in intraventricular gradient to 25 +/- 26 mmHg (p < 0.01) and a progressive decrease in intraventricular septum thickness (25 +/- 5 mm before treatment, 21 +/- 6 mm before hospital discharge, 17 +/- 3 mm at follow-up, p < 0.01). CONCLUSIONS: PTSMA of the intraventricular septum effectively relieves symptoms in selected patients with hypertrophic obstructive cardiomyopathy. The immediate decrease in intraventricular gradient is followed by a further decline at follow-up with a progressive reduction in the intraventricular septum thickness. PMID- 10806988 TI - Comparison of electrocardiographic criteria for diagnosis of left ventricular hypertrophy in hypertension: the MAVI study. AB - BACKGROUND: Standard electrocardiography (ECG) is a specific, but poorly sensitive tool for diagnosis of left ventricular (LV) hypertrophy. In a large population of subjects with hypertension we tested some standard ECG criteria in their sensitivity and specificity for LV hypertrophy. LV mass at echocardiography was the reference standard. METHODS: In the setting of the MAVI (MAssa Ventricolare sinistra nel soggetto Iperteso) study, the ECG and echocardiographic tracings of 947 hypertensive subjects were read blindly in a central office. RESULTS: Prevalence of LV hypertrophy at ECG was 0.6, 3.0, 4.8, 7.1, 11.1, 11.9 and 18.4%, respectively, using the following criteria: Wilson, typical strain, Romhilt-Estes score > or = 5 points, Gubner-Ungerleider, Sokolow-Lyon, Cornell voltage (S(V3)+R(avL) > 2.8 mV in men or 2.0 mV in women) and Perugia score (positivity of at least one of the following: S(V3)+R(aVL) > 2.4 mV in men or > 2.0 mV in women, a typical strain pattern, or a Romhilt-Estes point score >or = 5). Prevalence of LV hypertrophy at echocardiography ranged from 27.2% (LV mass > 125 g/m2) to 49.9% (LV mass > 51.0 g/m2.7). Using the latter gold standard, sensitivity and specificity of the above ECG criteria were 0.8 and 99.6% (Wilson), 3.8 and 97.9% (strain), 5.9 and 96.4% (Romhilt-Estes), 9.7 and 95.6% (Gubner-Ungerleider), 11.2 and 91.1% (Sokolow-Lyon), 15.2 and 91.4% (Cornell), and 22.2 and 85.4% (Perugia score). CONCLUSIONS: Sensitivity of traditional ECG criteria for LV hypertrophy in subjects with hypertension is poor. However, the combination of three highly specific criteria (Romhilt-Estes, LV strain and Cornell) in a cumulative score produces a rise in sensitivity without excessive deterioration of specificity, with a prevalence of LV hypertrophy at ECG of 18.4%. Traditional interpretation of ECG is valuable and should be reconsidered in the clinical work-up of subjects with hypertension. PMID- 10806989 TI - Evaluation of ischemia-reperfusion damage during coronary angioplasty. Electrocardiographic assessment and biochemical modifications in blood from the coronary sinus. AB - BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) constitutes a clinical model of reperfusion following a short period of ischemia connected to balloon inflation during the procedure. During the procedure some ischemic damage and oxidative injury related to free radical attack might occur. In the present study we investigated the extent of ischemic damage and some biochemical indexes of reperfusion damage in patients undergoing PTCA. METHODS: Twenty-five patients who underwent PTCA because of angiographically detected occlusion of the coronary artery were enrolled. Balloon inflation lasted from 30 to 60 s. ECG changes were monitored throughout the procedure and blood samples were taken from the coronary artery and coronary sinus before balloon inflation, and again from coronary sinus at the peak of ischemia, 2 and 10 min after reperfusion. RESULTS: During PTCA procedure angina pectoris appeared in 62.7% of patients, whereas ST-segment elevation was present in 87% of patients, regressing completely after balloon deflation. Plasma malonyldialdehyde, an index of lipid peroxidation, did not change; coenzyme Q10 (in its oxidized and reduced forms), vitamin E and beta carotene were also unchanged. Total antioxidant capacity and uric acid decreased upon reperfusion. CONCLUSIONS: Myocardial ischemia occurring during balloon inflation is brief and regresses completely after balloon deflation. Reperfusion following a short period of acute ischemia such as in PTCA does not constitute an oxidative event detectable through a common marker of lipid peroxidation nor does it alter the concentration of lipophilic antioxidants. It only lowers hydrosoluble antioxidants therefore representing a mild oxidative insult. PMID- 10806990 TI - Relationship between myocardial 123I-metaiodobenzylguanidine scintigraphic uptake and heart rate variability in patients with syndrome X. AB - BACKGROUND: We have recently demonstrated a striking impairment in cardiac uptake of 123I-metaiodobenzylguanidine (MIBG) in most patients with syndrome X. In this study we investigated the relationship between cardiac MIBG defects and cardiac autonomic activity in these patients. METHODS: MIBG myocardial scintigraphy and time-domain and frequency-domain heart rate variability (HRV) were compared in 11 syndrome X patients and 10 healthy controls. Cardiac MIBG uptake was assessed by the heart/mediastinum ratio and a cardiac MIBG uptake defect score (higher values = lower uptake). RESULTS: The heart/mediastinum ratio was lower (1.71 +/- 0.6 vs 2.19 +/- 0.3, p = 0.03) and MIBG uptake score higher (37.1 +/- 32 vs 4.0 +/- 2.5, p = 0.005) in syndrome X patients, whereas average HRV values did not differ between the two groups. However, while there were no correlations between MIBG uptake and HRV in controls, in syndrome X patients both the heart/mediastinum ratio and MIBG uptake score correlated significantly with two HRV parameters, specific for vagal activity: the square root of the mean squared differences of consecutive RR intervals (r = 0.73, p = 0.01, and r = -0.67, p = 0.02, respectively), and high frequency (r = 0.64, p = 0.03, and r = -0.74, p = 0.009, respectively). CONCLUSIONS; In patients with syndrome X, the impairment in cardiac MIBG uptake was associated with a reduction in HRV indexes mainly reflecting vagal modulation of sinus node, thus suggesting that a predominance of cardiac adrenergic activity may be present in those with abnormal cardiac MIBG scintigraphy. PMID- 10806991 TI - Relationship between baseline sympatho-vagal balance and the autonomic response to cardiac rehabilitation after a first uncomplicated myocardial infarction. AB - BACKGROUND: After a first uncomplicated myocardial infarction, cardiac rehabilitation increases the parasympathetic tone, in a direction linked to a reduced risk of sudden cardiac death. This change in sympatho-vagal balance may be related to other clinical variables. The aim of this study was to define the factors implicated in determining the autonomic response to cardiovascular rehabilitation after myocardial infarction. METHODS: In 55 patients (39-80 years) we evaluated the modulation of the autonomic profile induced by 8 weeks of rehabilitation: we analyzed the changes in pNN50 derived from time-domain analysis (deltapNN50) and in the low frequency/high frequency (LF/HF) ratio derived from autoregressive power spectral analysis (deltaLF/HF). A control group of 15 patients not undergoing rehabilitation was also studied. Variables considered at 4 weeks postinfarction and related to deltapNN50 and deltaLF/HF ratio were: age, site of myocardial infarction, previous thrombolysis, ejection fraction, stress test duration, baseline LF/HF ratio. RESULTS: Patients not undergoing rehabilitation did not change their autonomic profile. On the contrary, rehabilitation induced a higher vagal tone (pNN50 from 6.5 +/- 1.5 to 16.2 +/- 3.1; LF/HF ratio from 8.3 +/- 5.2 to 5.1 +/- 2.9, p < 0.05). Eleven patients (20%) had baseline LF/HF ratio exceeding the mean value by 1.5 SD (19.4 +/- 1.4): in this subgroup, pNN50 was very low. In these patients, rehabilitation increased pNN50 and decreased LF/HF ratio. Indeed, both deltapNN50 and deltaLF/HF ratio were significantly related to their baseline values (p < 0.001), even considering thrombolysis, site of myocardial infarction, age, and beta-blocker therapy. CONCLUSIONS: After a first uncomplicated myocardial infarction, sympatho vagal balance may be very disturbed in some patients, despite a preserved ventricular function, good exercise capability and beta-blockers. These patients should be encouraged to undergo rehabilitation, since the significant improvement in the parasympathetic tone may protect them against subsequent arrhythmic events. PMID- 10806992 TI - Transcatheter mechanical thrombus aspiration for stent thrombosis. AB - Currently, in-stent thrombosis is a rare but serious clinical event. The mechanical or pharmacological approach has not totally solved this problem. In this report we describe the treatment of in-stent thrombosis with a new device for mechanical thrombus aspiration. We used the Rescue catheter (Rescue Catheter System, Boston Scientific), a new 4.5F dual lumen monorail catheter that was able to break and aspirate thrombus without evidence of distal embolization. In this case the procedure was quickly performed with good angiographic results after mechanical aspiration and additional traditional coronary angioplasty. Moreover, the excellent clinical outcome confirmed the efficacy of the technique in the percutaneous treatment of this late complication of stent implantation. PMID- 10806993 TI - Novel mechanisms for antisense-mediated regulation of gene expression. PMID- 10806994 TI - Antisense oligonucleotides as a tool for gene functionalization and target validation. PMID- 10806995 TI - Molecular mechanisms of action of antisense drugs. AB - Given the progress reported during the past decade, a wide range of chemical modifications may be incorporated into potential antisense drugs. These modifications may influence all the properties of these molecules, including mechanism of action. DNA-like antisense drugs have been shown to serve as substrates when bound to target RNAs for RNase Hs. These enzymes cleave the RNA in RNA/DNA duplexes and now the human enzymes have been cloned and characterized. A number of mechanisms other than RNase H have also been reported for non-DNA like antisense drugs. For example, activation of splicing, inhibition of 5'-cap formation, translation arrest and activation of double strand RNases have all been shown to be potential mechanisms. Thus, there is a growing repertoire of potential mechanisms of action from which to choose, and a range of modified oligonucleotides to match to the desired mechanism. Further, we are beginning to understand the various mechanisms in more detail. These insights, coupled with the ability to rapidly evaluate activities of antisense drugs under well controlled rapid throughput systems, suggest that we will make more rapid progress in identifying new mechanisms, developing detailed understanding of each mechanism and creating oligonucleotides that better predict what sites in an RNA are most amenable to antisense drugs of various chemical classes. PMID- 10806996 TI - Two problems in antisense biotechnology: in vitro delivery and the design of antisense experiments. AB - Antisense oligonucleotides are invaluable reagents for the specific downregulation of gene expression. In the absence of a carrier, charged oligonucleotides (e.g., phosphorothioates) can interact with a large number of cell surface proteins, but tend to be internalized into the endosomal/lysosomal compartment. However, they can be successfully delivered to the nuclei of diverse cell types via the use of a wide variety of reagents, including cationic lipids, and cationic polyamines. Over the past several years, a more general understanding of the rules governing the interpretation of data derived from antisense experiments has been reached. These are discussed with emphasis on how to avoid some of the confounding features of this important, emerging technology. PMID- 10806997 TI - Importance of nucleotide sequence and chemical modifications of antisense oligonucleotides. AB - The antisense approach is conceptually simple and elegant; to design an inhibitor of a specific mRNA, one needs only to know the sequence of the targeted mRNA and an appropriately modified complementary oligonucleotide. Of the many analogs of oligodeoxynucleotides explored as antisense agents, phosphorothioate analogs have been studied the most extensively. The use of phosphorothioate oligodeoxynucleotides as antisense agents in various studies have shown promising results. However, they have also indicated that quite often, biological effects observed could be solely or partly non-specific in nature. It is becoming clear that not all phosphorothioate oligodeoxynucleotides of varying length and base composition are the same, and important consideration should be given to maintain antisense mechanisms while identifying effective antisense oligonucleotides. In this review, I have summarized the progress made in my laboratory in understanding the specificity and mechanism of actions of phosphorothioate oligonucleotides and the rationale for designing second-generation mixed-backbone oligonucleotides. PMID- 10806998 TI - A review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides. PMID- 10806999 TI - Oligonucleotide therapeutics for hematologic disorders. AB - During the last decade, the catalogue of known genes responsible for cell growth, development, and neoplastic transformation has expanded dramatically. Attempts to translate this information into new therapeutic strategies for both hematologic and non-hematologic diseases have accelerated at a rapid pace as well. Inserting genes into cells which either replace, or counter the effects of disease causing genes has been one of the primary ways in which scientists have tried to exploit this new knowledge. Strategies to directly downregulate gene expression have developed in parallel with this approach. The latter include triple helix forming oligonucleotides (ODN) and 'antisense' ODN. The latter have already entered clinical trials for a variety of disorders. In this monograph, we review the use of these materials in the treatment of hematologic diseases, particularly myelogenous leukemias. Problems and possible solutions associated with the use of ODN will be discussed as well. PMID- 10807000 TI - Human Bcl-2 antisense therapy for lymphomas. PMID- 10807001 TI - Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides. AB - Immune stimulation has been widely recognized as an undesirable side effect of certain antisense oligodeoxynucleotides (ODN) which can interfere with their therapeutic application. It is now clear that these dose-dependent immune stimulatory effects primarily result from the presence of an unmethylated CpG dinucleotide in particular base contexts ('CpG motif). The sequence-specific immune activation is not just an experimental artifact, but is actually a highly evolved immune defense mechanism whose actual 'goal' is the detection of microbial nucleic acids. In contrast to vertebrate DNA, in which CpG dinucleotides are 'suppressed' and are highly methylated, microbial genomes do not generally feature CpG suppression or methylation [1]. Immune effector cells such as B cells, macrophages, dendritic cells, and natural killer cells appear to have evolved pattern recognition receptors (PRR) that by binding the microbe restricted structure of CpG motifs, trigger protective immune responses. Although the specific immune activation appears to have a variety of potential therapeutic applications, it is generally undesirable in antisense ODN. Immune stimulation may be avoided in antisense oligos by the selection of CpG-free target sequences, by the use of ODN backbones that do not support immune stimulation, or by selective modifications of the cytosine in any CpG dinucleotides. PMID- 10807002 TI - 2'-carbohydrate modifications in antisense oligonucleotide therapy: importance of conformation, configuration and conjugation. AB - The 2'-position of the carbohydrate moiety has proven to be a fertile position for oligonucleotide modifications for antisense technology. The 2'-modifications exhibit high binding affinity to target RNA, enhanced chemical stability and nuclease resistance and increased lipophilicity. All high binding affinity 2' modifications have C3'-endo sugar pucker. In addition to gauche effects, charge effects are also important in determining the level of their nuclease resistance. Pharmacokinetic properties of oligonucleotides are altered by 2'-conjugates. For certain modifications (e.g., 2'-F), the configuration at the 2'-position, arabino vs. ribo, determines their ability to activate the enzyme RNase H. PMID- 10807003 TI - Oligonucleotide N3'-->P5' phosphoramidates as potential therapeutic agents. AB - Uniformly modified nucleic acids analogues, oligonucleotide N3'-->P5' phosphoramidates, containing 3'-amino instead of 3'-hydroxyl nucleosides, were synthesized and studied. These compounds form very stable duplexes with complementary native phosphodiester DNA and exceptionally stable duplexes with RNA strands. Increases in duplex melting temperature, deltaTm, relatively to their phosphodiester counterparts, reaches 2.9-3.5 degrees C per modified nucleoside. Moreover, the phosphoramidate compounds form extremely stable triple stranded complexes with single or double stranded DNA oligomers under near physiological salt and pH conditions. Melting temperatures of these triplexes usually exceed that of the isosequential phosphodiester counterparts by up to 35 degrees C. For 11-15-mers 2'-deoxyphosphoramidates are structurally and functionally similar to the native RNA molecules and thus can be used as RNA decoys. They are resistant to enzymatic digestion by nucleases both in vitro and in vivo. Oligonucleotide phosphoramidates apparently are cell permeable, and they have a good bioavailability and biodistribution, while being non-toxic in mice at therapeutically relevant doses. Duplexes of the several studied phosphoramidates with complementary RNA strands apparently are not substrates for RNase H in vitro. Despite that, these compounds exerted high sequence-specific antisense activity in various cell lines and in SCID mice. The observed in vitro lack of RNase H recognition of the RNA:phosphoramidate duplexes may result in better specificity in biological activity of these compounds relative to RNase H inducing oligonucleotides. Experimental results also indicate that oligonucleotide phosphoramidates can be used as efficient and specific modulators of gene expression by an antigene mechanism of action. Finally, the oligo-2' deoxyphosphoramidate double stranded complexes can structurally mimic native RNA complexes, which could be efficiently and specifically recognized by the RNA binding proteins, such as HIV-1 Rev and Tat. PMID- 10807004 TI - Morpholino antisense oligomers: the case for an RNase H-independent structural type. AB - RNase H-competent phosphorothioates (S-DNAs) have dominated the antisense field in large part because they offer reasonable resistance to nucleases, they afford good efficacy in cell-free test systems, they can be targeted against sites throughout the RNA transcript of a gene, and they are widely available from commercial sources at modest prices. However, these merits are counterbalanced by significant limitations, including: degradation by nucleases, poor in-cell targeting predictability, low sequence specificity, and a variety of non antisense activities. In cell-free and cultured-cell systems where one wishes to block the translation of a messenger RNA coding for a normal protein, RNase H independent morpholino antisense oligos provide complete resistance to nucleases, generally good targeting predictability, generally high in-cell efficacy, excellent sequence specificity, and very preliminary results suggest they may exhibit little non-antisense activity. PMID- 10807005 TI - Antisense properties of peptide nucleic acid. AB - Peptide nucleic acid (PNA) is a nucleic acid mimic in which the deoxyribose phosphate backbone has been replaced by a pseudo-peptide polymer to which the nucleobases are linked. PNA-oligomers can be synthesized in relatively large amounts, are highly stable in biological environments, and bind complementary DNA and RNA targets with remarkably high affinity and specificity. Thus PNA possesses many of the properties desired for a good antisense agent. Until recently, limited uptake of PNA into cells has been the major obstacle for applying PNA as an antisense agent in cell cultures and in vivo. Here, the antisense properties of PNA in vitro and in vivo will be reviewed. In particular, we will focus on recent observations indicating that PNA equipped with or without various uptake moieties may function as an efficient and gene-specific inhibitor of translation in Escherichia coli and in certain mammalian cell types. PMID- 10807006 TI - Conformationally restricted carbohydrate-modified nucleic acids and antisense technology. AB - The study of conformationally restricted carbohydrate modified nucleic acids has given new insights into the concept of the antisense technology. We learned to understand the structural requirements of a modified nucleic acid to function as steric blocker for RNA. Several of the physicochemical and conformational factors influencing duplex stabilization are analyzed with respect to their relative importance for the antisense field. PMID- 10807007 TI - Triple helix formation and the antigene strategy for sequence-specific control of gene expression. AB - Specific gene expression involves the binding of natural ligands to the DNA base pairs. Among the compounds rationally designed for artificial regulation of gene expression, oligonucleotides can bind with a high specificity of recognition to the major groove of double helical DNA by forming Hoogsteen type bonds with purine bases of the Watson-Crick base pairs, resulting in triple helix formation. Although the potential target sequences were originally restricted to polypurine polypyrimidine sequences, considerable efforts were devoted to the extension of the repertoire by rational conception of appropriate derivatives. Efficient tools based on triple helices were developed for various biochemical applications such as the development of highly specific artificial nucleases. The antigene strategy remains one of the most fascinating fields of triplex application to selectively control gene expression. Targeting of genomic sequences is now proved to be a valuable concept on a still limited number of studies; local mutagenesis is in this respect an interesting application of triplex-forming oligonucleotides on cell cultures. Oligonucleotide penetration and compartmentalization in cells, stability to intracellular nucleases, accessibility of the target sequences in the chromatin context, the residence time on the specific target are all limiting steps that require further optimization. The existence and the role of three stranded DNA in vivo, its interaction with intracellular proteins is worth investigating, especially relative to the regulation of gene transcription, recombination and repair processes. PMID- 10807008 TI - Unintended immunomodulation: part I. Effects of common clinical conditions on cytokine biosynthesis. AB - Cytokines are low molecular weight proteins that act in an autocrine, paracrine and endocrine fashion to regulate and integrate immune effector cell function. Cytokine production is tightly controlled by a complex network of co-stimulatory and feedback loops. The systemic concentrations of some cytokines, most notably tumor necrosis factor and various interleukins, correlate with the extent of inflammation, and the severity of critical illness and patient outcome. Thus, cytokine expression is often monitored and/or manipulated as a therapeutic target in studies of sepsis and other inflammatory conditions. Unfortunately, some therapies designed to modify cytokine response have failed to improve outcomes in sepsis, and some of these therapies have actually been harmful. Several common clinical conditions, as well as, therapeutic interventions significantly influence cytokine expression. Furthermore, the magnitude and extent of these effects may be greater than those produced by immunomodulating therapies. In contrast, other conditions may not produce clinically significant changes in cytokine expression, and must simply be considered when interpreting studies designed to determine the effects of immunomodulation. Some conditions may even result in changes in the inflammatory response and may thus add to the inflammatory burden of a critically ill patient. This review provides intensivists and other clinicians with an overview of the effects of altered physiologic conditions on cytokine expression. This information is important so that studies measuring cytokines can be correctly interpreted and clinical circumstances in which cytokine manipulation is undesirable can perhaps be avoided. PMID- 10807009 TI - Unintended immunomodulation: part II. Effects of pharmacological agents on cytokine activity. AB - Cytokines are proteins that are produced by immune and non-immune cells, and they function as mediators to facilitate cellular communication. Their production is regulated by a complex network of co-stimulatory and feedback loops that responds to a variety of stimuli. Several pharmacological agents have been found to alter systemic concentrations and/or the activity of different cytokines via a variety of mechanisms, including changes in biosynthesis, secretion, and/or stability. Many of the agents that modulate cytokine levels commonly are used in the management of critically ill patients. Catecholamines for example, have been found to promote the secretion of anti-inflammatory cytokines, and, therefore, may alter acute inflammatory processes such as sepsis. Antimicrobials have multiple effects on cytokine production, either secondary to the release of endotoxins from gram-negative bacteria or via direct activity on cytokine expression at the transcriptional and/or post-transcriptional level. Pentoxifylline has multiple effects on the immune system, but inhibition of pro inflammatory cytokine release predominates. The reminder of the known drug cytokine interactions and their effect on the inflammatory process are discussed. Information on the pharmacodynamic effect of drugs is limited, and our understanding of the clinical significance of these observations awaits further investigation. This review was designed to provide intensivists and other clinicians with useful information regarding the effect of medications on cytokine activity. It is also intended to help researchers and clinicians to optimize the design of studies of pharmacotherapeutic modulation of cytokines and to avoid the use of some agents in clinical circumstances in which cytokine manipulation is undesirable. PMID- 10807010 TI - Neutrophil apoptosis is delayed in patients with inflammatory bowel disease. AB - Delayed neutrophil apoptosis is a feature of persistent acute inflammation. Neutrophil-mediated damage has been shown to be associated with the development of inflammatory bowel disease (IBD). Persistence of these cells both at the colonic site and circulation may further contribute to IBD. The aims of this study were to determine whether neutrophils isolated from IBD patients delay apoptosis and to investigate possible mechanisms involved in this delay. We studied 20 patients with IBD, 13 with Crohn's disease, and 7 with ulcerative colitis, all of whom were undergoing intestinal resection for symptomatic disease. Seventeen patients undergoing elective resection of colon cancer acted as operative controls. Systemic, mesenteric arterial, and mesenteric venous blood was harvested. Neutrophils isolated from patients with IBD showed decreased spontaneous apoptosis compared to cancer patients. Mesenteric venous serum of IBD patients contributed to this delay, which contained higher concentrations of interleukin-8 (IL-8). Pro-caspase 3 expression was also reduced in IBD neutrophils, which may contribute to decreased spontaneous and Fas antibody induced apoptosis. Neutrophil apoptosis may be altered in Crohn's disease and ulcerative colitis through release of anti-apoptotic cytokines and altered caspase expression. The alterations in cell death mechanisms may lead to persistence of the inflammatory response associated with IBD. PMID- 10807011 TI - Hemodynamic effects of hypertonic hydroxyethyl starch 6% solution and isotonic hydroxyethyl starch 6% solution after declamping during abdominal aortic aneurysm repair. AB - Fluid resuscitation with hypertonic hydroxyethyl starch solutions (HES) is effective in haemorrhagic shock due to the rapid mobilisation of fluids into the intravascular compartment. Declamping of the abdominal aorta with acute redistribution of blood into the vessels of the lower body half causes declamping induced hypotension. Usually large amount of fluids or vasopressors are necessary to restore hemodynamic stability. Therefore, infusion of a hypertonic colloid solution may be an attractive option to achieve hemodynamic stability. This study was conducted to determine the amount of fluid of either hypertonic HES (HES 6%;7.2% NaCl) or isotonic HES (HES 6%;0.9% NaCl) needed to attain best wedge pressure (PCWP) cardiac index (CI) relation after declamping. Thirty-two high risk patients undergoing elective abdominal aneurysm resection were enrolled in a prospective, randomised, double blinded study. The individual optimised PCWP/CI relation was determined after induction of anaesthesia. After declamping, both solutions were titrated in small boluses of 100 mL until the previously determined best wedge was reached. The amount of fluid after declamping was significantly reduced in the hypertonic HES- group 162 mL vs. 265 mL in the control group (P < 0.05). Resuscitation time was shortened, and cardiac index was slightly higher in the treatment group. The use of hypertonic HES-solution after aortic declamping led to a significant reduction of fluids necessary to attain optimised PCWP/CI relation. In this clinical trial with moderate blood loss in high-risk patients, hypertonic HES applied in a titrated fashion restored hemodynamic stability faster and without volume overload. PMID- 10807012 TI - Interleukin-1beta induces complement component C3 and IL-6 production at the basolateral and apical membranes in a human intestinal epithelial cell line. AB - In previous studies, stimulation of cultured enterocytes with IL-1beta resulted in production of IL-6 and complement component C3. The cellular mechanisms of these responses in the enterocyte are not fully understood. We tested the hypothesis that IL-1beta-induced C3 and IL-6 production is differentially regulated at the apical and basolateral membranes of the enterocyte. Caco-2 cells (a transformed human colonic carcinoma cell line) were grown in a 2-chamber system to full differentiation. The cells were treated with IL-1beta either at the apical or basolateral membrane, and C3 and IL-6 mRNA levels and release of C3 and IL-6 into the apical and basal chambers were determined. The release of C3 was greatest into the basal chamber regardless of whether the cells were stimulated at the apical or basolateral membrane. In contrast, the production of IL-6 was greatest at the cell membrane that was stimulated with IL-1beta. Stimulation of the Caco-2 cells with IL-1beta resulted in increased mRNA levels for C3 and IL-6 with no major differences noted when the cells were treated at the apical or basolateral membrane. The results suggest that enterocyte production and release of at least some acute phase proteins and cytokines are differentially regulated at the apical and basolateral membrane of the enterocyte after stimulation with IL-1beta. PMID- 10807013 TI - Alterations in tissue glucose uptake during the hyperglycemic and hypoglycemic phases of sepsis. AB - The purpose of the present study was to characterize the alterations in tissue glucose uptake during the hyperglycemic, euglycemic, and hypoglycemic phases of peritonitis. Rats had vascular catheters implanted, and sepsis was induced by cecal ligation and puncture. Rates of whole-body glucose appearance (Ra), disappearance (Rd), and metabolic clearance (MCR) were determined by the constant infusion of 3H-glucose, and in vivo glucose uptake (Rg) by individual tissues was assessed by using 14C-deoxyglucose. During the hyperglycemic phase of sepsis (2 h), glucose Ra and Rd were increased, but glucose MCR was unaltered. In contrast, during the euglycemic phase (6 h), the sepsis-induced increase in glucose Ra and Rd was associated with an elevation in the MCR. Finally, during the hypoglycemic phase (24 h), sepsis decreased glucose Ra and Rd and the glucose MCR. The sepsis induced changes in Rg for skeletal muscle and adipose tissue mimic those seen for the whole body at each time point. Rg for skin and intestine was elevated at 2 h and 6 h but was not different from control values at 24 h. In contrast, the Rg for liver, lung, and spleen was increased at all 3 time points. In a second study, there was no difference in Rg for any tissue between 2-h septic rats and control animals in which blood glucose and insulin levels were artificially elevated to the same degree. In a third study, the prevailing glucose and insulin levels in control animals were decreased, by injection of the gluconeogenic inhibitor 3-mercaptopicolinic acid, to levels seen in 24-h septic rats. There was no difference in the Rg for muscle and adipose tissue between 24-h septic rats and hypoglycemic insulinopenic control animals. However, the Rg for liver, lung, and spleen remained elevated in 24-h septic rats, compared with hypoglycemic insulinopenic control values. These data indicate that the increased tissue glucose uptake observed during the early phase of sepsis is a consequence of concomitant changes in plasma glucose and insulin. In contrast, during the euglycemic and hypoglycemic stages of sepsis, glucose uptake in macrophage-rich tissues remains elevated and is independent of changes in glucose and insulin. PMID- 10807014 TI - Protection against endotoxemia in rats by a novel tetrahydrobiopterin analogue. AB - We studied the effects of a novel pterin antagonist of NO synthase, the 4-amino analogue of tetrahydrobiopterin (4-ABH4), in a rat model of endotoxic shock and compared its properties with those of N(G)-monomethyl L-arginine (L-NMMA). Treatment with a bolus dose of 4-ABH4 at 2 h after LPS challenge significantly improved the 6-day survival rate, compared with the controls treated with saline. L-NMMA treatment did not significantly influence the survival rate. This bolus treatment, using either compound, had no effect on the plasma nitrite + nitrate or plasma IL-6 levels. The continuous infusion of 4-ABH4 efficiently suppressed the enhanced calcium-dependent/independent NO synthase activities induced by endotoxin in lung homogenates and completely suppressed the increase in plasma nitrite + nitrate caused by endotoxin at 5 h, with no significant difference compared with the L- NMMA treatment. Treatment of RAW264.7 murine macrophages with 4-ABH4 but not with L-NMMA suppressed endotoxin-induced tumor necrosis factor-alpha release by the cells, whereas nitrite in the supernatant decreased in a dose-dependent fashion in both assay systems. Our data show that 4-ABH4, an inhibitor of inducible NO synthase, significantly improves survival in a rat model of endotoxic shock when administered in a bolus dose that does not reduce plasma total nitrite + nitrate levels. Because we observed no overt signs of toxicity and no influence on organ-specific tetrahydrobiopterin levels, we conclude that the novel compound 4-ABH4 is a promising drug candidate for protection against endotoxin-related mortality. PMID- 10807015 TI - Roles of cyclooxygenase-2 in tissue injury during hemorrhagic shock. AB - Prostaglandins (PGs), which play a pivotal role in the cytokine network, have been implicated in the pathophysiology of circulatory shock, but the precise role of cyclooxygenase-2 (COX-2) in circulatory shock is still not known. This study investigated whether or not COX-2 affects liver and bowel injury during hemorrhagic shock (HS). Male Sprague-Dawley rats were subjected to decompensated HS followed by resuscitation. Besides the time course of tissue injury and mRNA expression of COX-2 in the liver and bowel during HS, we investigated the effect of N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide (NS398), a specific inhibitor of COX-2, on injury to these organs. The liver injury, evaluated by plasma aminotransferase levels and histology, was evident at the end of the shock period (Shock-0h) and had significantly increased 1 h after the start of resuscitation (Shock-1h) (P < 0.01). The bowel injury, assessed by histological findings in the terminal ileum, was scarcely detectable at Shock-0h but became noticeable at Shock-1h (P < 0.01). COX-2 mRNA expression was up-regulated in the liver during HS whereas it did not change in the bowel. NS398 treatment significantly exacerbated both liver and bowel injury. These lines of evidence suggest that COX-2-derived PGs provide protection against HS-induced liver and bowel injury. PMID- 10807016 TI - Endotoxin-induced mesenteric microvascular changes involve iNOS-derived nitric oxide: results from a study using iNOS knock out mice. AB - The administration of endotoxin alters intestinal blood flow, increases nitric oxide (NO) production, and induces gut barrier dysfunction. Thus, we investigated the hypothesis that microvascular reactivity and permeability of the mesenteric vascular bed are altered to a lesser degree in iNOS knock out (iNOS -/-) mice than their wild-type (iNOS +/+) litter mates after an endotoxin challenge. To test this hypothesis, we compared the microvascular response of iNOS knockout (iNOS -/-) mice after a topical or systemic endotoxin challenge against that of their wild-type litter mates (iNOS +/+). Intravital microscopy was used to measure arteriolar diameter and postcapillary venular permeability in the mouse ileum. Both parameters were determined by computer-assisted image analysis. Diameter was measured in A1, A2, and A3 arterioles (1, 2, 3 = rank of deployment). Changes in microvascular permeability were measured from changes in interstitial fluorescence caused by extravasation of fluorescein isothiocyanate (FITC)-dextran 150 (molecular weight = 150 kDa) and expressed as changes in integrated optical intensity (IOI). In the first series of experiments, endotoxin (100 microg/mL) was applied topically to the ileal segment. In the second series, endotoxin (10 mg/kg) was administered intraperitoneally (i.p.). Administration of topical or i.p.. endotoxin caused vasoconstriction and was associated with an early increase in permeability in both iNOS +/+ and -/- mice, although over time the responses of the iNOS -/- and iNOS +/+ mice diverged. Twenty minutes after topical endotoxin, the increase in permeability in iNOS -/- mice had reached a plateau whereas it continued to increase in the iNOS +/+ mice, such that at 80 min post-topical endotoxin, IOI was 27+/-7 in iNOS -/- vs. 39+/-5 in iNOS +/+ (P < 0.05). A similar permeability response was observed after i.p.. endotoxin, where the increase in post-capillary venular permeability was greater in the iNOS +/+ mice (P < 0.05). Both iNOS -/- and iNOS +/+ mice had a similar transient vasoconstrictive response after topical endotoxin challenge (reduction in A2 arteriolar diameters by -17+/-4% vs. -24+/-7%), with return to baseline values by 60-80 min post-endotoxin challenge. The iNOS +/+ but not the iNOS -/- mice manifested a secondary vasodilatory response that persisted throughout the experimental period. The arteriolar vasoreactive response of the iNOS -/- and iNOS +/+ mice to i.p.. endotoxin was similar to that of topical endotoxin, but of a lesser magnitude. In conclusion, the similarity in effects between topical and systemic endotoxin indicates that endotoxin causes microvascular dysfunction in the gut by directly on the microcirculation. In addition, our data suggest that NO production by iNOS is involved in the microvascular alterations associated with gut barrier dysfunction. PMID- 10807017 TI - Proinflammatory cytokines increase in sepsis after anti-adhesion molecule therapy. AB - Cytokine mediators and leukocyte-endothelial cell adhesion molecules are critical and interdependent components of the acute inflammatory response in sepsis. We hypothesized that the administration of monoclonal antibodies to intercellular adhesion molecule-1 (CD54) or E- and L-selectin (CD62E/L) would decrease serum levels of the proinflammatory cytokines interleukin-1beta (IL-1), IL-6, and IL-8 and tumor necrosis factor receptor (TNFR-1) in baboons during sepsis. Adult male baboons received infusions of 1 x 10(9) colony forming units (CFU)/kg heat-killed Escherichia coli (E. coli) followed 12 h later by live E. coli (1 x 10(10) CFU/kg). At the time of live bacterial infusion, six septic animals were treated with a monoclonal antibody to CD54 and six with an antibody to CD62E and L (1 mg/kg). Eight untreated septic animals served as controls. Sequentially drawn serum samples were assayed for IL-1, IL-6, IL-8, and TNFR-1 using enzyme-linked immunoassay (ELISA). Data were compared using Mann-Whitney U tests and Chi-square analyses. Median survival was decreased in both treatment groups compared to controls (P < 0.05). Peak IL-1 level was higher than controls in septic animals treated with anti-CD54 but not anti-CD62E/L (P < 0.05, P = NS, respectively). Elevations in IL-6, IL-8, and TNFR-1 were increased and prolonged in both antibody treated groups compared to controls (P < 0.05). These results provide the first in vivo evidence that leukocyte-endothelial adhesion molecules CD54 and CD62E/L regulate cytokine production in sepsis. PMID- 10807018 TI - Meretricious mensuration. PMID- 10807019 TI - Public health in the undergraduate medical curriculum--can we achieve integration? AB - Public health is widely regarded by medical students as peripheral or even irrelevant to the acquisition of clinical knowledge and skills. This paper attempts to set out some of the reasons for this, to encourage innovative approaches to integrating public health with clinical teaching and to offer a theoretical framework of integrated public health education for curriculum development and evaluation. The points of convergence between public health and clinical practice should not be regarded as self-evident. A practical demonstration of the application of public health principles to clinical problem solving may be the most effective means of overcoming resistance. Almost anywhere that clinical services are provided is suitable for this purpose. Community clinics, health centres or general practices have obvious appeal but acute hospitals have important advantages arising from students' preoccupation with clinical medicine. The main aim of integrated public health teaching is to facilitate the students' acquisition of knowledge, skills and attitudes that promote the effective application of public health approaches to clinical practice. The interrelationships between clinical practice and public health may be represented in the form of a grid. The vertical headings are the clinical skills that relate to the different stages of the natural history of disease -- from the pre-disease state through diagnosis, treatment and follow up. The horizontal headings describe four key public health dimensions: epidemiology, behaviour/lifestyle, environment and health policy. The text in the boxes suggests appropriate topics for discussion. The grid is also potentially useful for course documentation and content evaluation. PMID- 10807020 TI - The Critical Care Research Network: a partnership in community-based research and research transfer. AB - The objectives of this study were to present a short history of the Critical Care Research Network (CCR-Net), describe its approach to health services research and to summarize completed and current research projects. In doing this, we explored the question is this research network accomplishing its goals? We reviewed the medical literature to identify studies on similar types of Networks and also the evidence supporting the methodology used by CCR-Net to conduct research using MEDLINE, HEALTHSTAR, CINAHL and the keywords network and health care or healthcare, benchmarking and health care or healthcare, and research transfer or research utilization. We also reviewed the bibliographies of retrieved articles and our personal files. In addition, we summarized the results of studies conducted by CCR-Net and outlined those currently in progress. A review of the literature identified studies on two similar networks that appeared to be succeeding. In addition, the literature was also supportive of the general process used by CCR-Net, although the level of evidence varied. Finally, the studies conducted to date within CCR-Net follow the suggested methodology. At the time of this preliminary communication CCR-Net appears to have adopted a valid approach to health services research within the area of Critical Care Medicine. Further direct evidence is required and appropriate studies are planned. PMID- 10807021 TI - Effect of case managers with a general medical patient population. AB - The objectives of this study were to evaluate the effect of inpatient case management (CM) on a general medical service and to determine if a prediction rule, identifying patients likely to need discharge planning services, could define a subset of patients for whom CM would be most effective. We hypothesized that CM would have greatest impact on patients predicted to be at highest risk of needing discharge planning to arrange for post-discharge medical services. We carried out a prospective controlled study. Six general medicine teams from a 600 bed urban teaching hospital were randomly assigned to CM (n = 4) or standard care (SC) (n = 2). Number of patients = 302 (207 CM; 95 SC). Case managers participated in daily physician team rounds and coordinated discharge planning for CM patients; SC patients received discharge planning from staff nurses or discharge planners when requested by physicians. The outcomes measured were deviation from the hospital length of stay (LOS) expected for a patient's diagnosis, patient satisfaction and non-acute medical service utilization during the month after discharge. Overall, patients from CM and SC teams did not differ in their deviation from expected LOS, post-discharge medical service utilization and patient satisfaction. However, after stratifying patients by their predicted need for post-discharge medical services, only patients in the 'high risk' category had a significantly shorter LOS under CM (2.9 days shorter than SC patients; P = 0.02). We concluded that, in this study, the effect of case managers on a general medical service was limited to shortening LOS only among a stratum of high risk patients. PMID- 10807022 TI - Tailor-made quality systems in healthcare. AB - At present many healthcare organizations are being pressed by national governments or client groups to develop and implement quality systems. Unfortunately, not much is known about the development and implementation of these systems in healthcare organizations. There still are definition questions to be answered and it is not clear in which way a quality system can be outlined and which outline is most effective in a particular situation. In practice, many healthcare organizations adopt ideas from industrial quality management and develop ISO 9000 based quality systems, which are only useful in relatively stable situations. In this paper a typology of healthcare processes is developed on the basis of input and throughput characteristics, following the literature on professional quality systems and service systems. The typology is tested and illustrated by a few case studies. From this typology, on the basis of logic, suggestions have been deduced for tailor-made quality systems in a general hospital setting. The main point of these systems is an examination, treatment and nursing plan with a timescale. Control is executed on the basis of performance indicators for groups of patients. PMID- 10807023 TI - Identifying meaningful intra-individual change standards for health-related quality of life measures. AB - Although numerous measures have been developed for the evaluation of health related quality of life (HRQoL), strategies for identifying meaningful intra individual change in these measures have not kept pace with instrument development. As a result, clinical trial researchers, quality assurance assessment teams and practising clinicians are without established standards to evaluate individual patient change in HRQoL measures as improved, stable or declined. This article reviews and critiques the methods that have been applied to establish intra-individual HRQoL change standards. These methods include within-person and between-persons anchor-based studies, as well as distribution based techniques using the effect size, the standard error of measurement, the mean squared error or individual slope coefficients derived from hierarchical linear modelling. Practical approaches to improving and advancing HRQoL change evaluations that enhance the interpretation of intra-individual change are provided. Two future methodological challenges in this area of HRQoL research are examined: (1) the development of individual change standards for generic HRQoL measures; and (2) the incorporation of individual clinical assessments into the process for establishing significant intra-individual change standards. PMID- 10807024 TI - Quality end-of-life care. PMID- 10807025 TI - The relationship of impending death and age category to treatment intensity in the elderly. AB - The purpose of this study was to isolate and quantify the age-related treatment intensity differential in elderly clients (n = 278) with functional disability in one managed care setting. For those who died during the study, treatment intensity changes for the last year and month of life were examined. The subjects were categorized into four age groups: 75-79 years; 80-84 years; 85-89 years and 90 + years, and a treatment intensity index was calculated for each group using a ratio of actual to expected costs. Indices of overall costs and cost per day for all clients, and also indices for the year and month prior to death for the deceased clients were calculated. The results clearly show that for all clients, the oldest age group was treated less intensively than the youngest age group. For the deceased clients, the older age group was treated less intensively than the youngest age group in the last year and month of life but, for all age groups, the intensity of treatment increased during the last month of life. PMID- 10807026 TI - Patients attending a new drug clinic in 1990 and 1995: characteristics and outcome. PMID- 10807027 TI - Community in gay male experience and moral discourse. AB - The three sociological conceptions of community are outlined: the imagined community, community as friendship, and community as local organizations or groups. The extent to which gay men in New York City experience a gay community in each of these ways is investigated by means of in-depth interviews. The type of moral discourse utilized by the gay men interviewed is also discussed. Finally, gay men in New York City are compared to the new Russian Jewish community in Brooklyn as to which is more "postmodern." PMID- 10807028 TI - Part 1: HIV as'the line in the sand'. AB - There is a growing rift between HIV-positive and HIV-negative gay men, which finds expression in social, economic, structural and political divisiveness that, if not resolved, may 'kill' the "gay liberation movement." While disasters generally tend to create organizational solidarity, the AIDS crisis has operated in reverse, spawning a variety of competitive AIDS service organizations, alienating seropositive gays from the mainstream gay community, and in turn disenfranchising seronegative gay men as human and financial resources are redirected toward persons living with HIV and AIDS. Serostatus has become a social marker of societal status, operating in a bimodal discriminatory manner. Seronegative gay men experience discrimination from within the gay community as funding for and services to this sector diminish. Seropositive gay men (and the organizations that provide for some of their needs) have culturally, economically and socially dismissed the socio/psychological needs of seronegative gay men (survivor guilt, safer sex education, etc.) in favour of providing social and resource-based services to seropositive gay men. As the disparities in service and advocacy increase, the social distance between the gay movement and the AIDS movement correspondingly increases. If this trend continues, the social gap will serve further to push HIV-positive and HIV-negative gay men into polarized camps, resulting in a wider separation of the gay movement and the AIDS movement. The stigmatization of HIV-positive people will subsequently increase both within and outside the gay movement, and any ability to present a unified Gay Liberation front will correspondingly diminish. Additionally, the emergent notion within and without the gay communities that to be gay is to be HIV-positive will solidify. This will (a) further stigmatize all gay men in the eyes of the non-gay population, and (b) exacerbate the rift between HIV-positive and HIV-negative gay men within the gay community, reversing the stigma of HIV such that to be HIV negative will be a marker of non-gay identity. In short, seropositivity will become the defining element of gayness. PMID- 10807029 TI - Part 2: Fear of contagion, fear of intimacy. AB - In this second part of the trilogy, I review the concepts of panic, the Theory of Cognitive Dissonance, and how internally inconsistent opinions and attitudes can be made consistent (or consonant). The theory explains, in some measure, how AIDS has been socialized into our thinking about identity, and goes beyond a medical condition. The pervasive identification of gay men with HIV and AIDS has resulted for many in an over-identification with fears of contagion and on a societal level in a fear of all gays as pools of contagion. The conversion of dissonance to consonance has taken many forms; within the gay community it has resulted in the rejection of the "100% safe-100% of the time" safe-sex message, and the adoption (for many) of a new form of deviant label-someone who is not in conformity with the social norm of gay community sexual behavior. However, we shall see that this so-called norm is a sham-that many gay men do not, as a rule, practice safe(r) sex on a consistent basis. This information indicates that the educational efforts of the last decade have at best lost their potency, and at worst were less than efficacious to begin with. The dissonant messages have also informed both the construction of the gay community and its interpretation of what it means to be gay. The result has been a tri-lateral perception of HIV and AIDS as either a medical, political or a social phenomenon. This fragmented understanding has exacerbated the already polarized ASOs and GSOs in that each has determined its ideology based on a particular interpretation of HIV and AIDS. This polarization has been operationalized by the GSOs and ASOs primarily in the manner by which they define their target markets, and more importantly, in the manner by which they exclude certain gays from participation. At the extreme, some gay men feel entirely left out of the community, and are consequently unable to convert their dissonance regarding being gay into consonance, if only by developing some associational ties with the community. The central question of the sustainability of the gay movement is thus partly answered by restating the nature of the fractures in the community. Kiesler's determinants regarding change relate directly to the sustainability question-can GSOs and ASOs, given their pluralistic ideologies and constituencies, break free of the constraints that are posed by these determinants? Would the adherents and conscious constituents defect from their organizations, and form new ones (thus reifying the fractures that already exist)? On the other hand, is there a sense of community and identity that will function as a bonding agent to encourage coalition building and social reorganization? The matter may turn on the issue of selective rewards: can a coalition of ASOs and GSOs provide staff, volunteers and clients sufficient motivation for making the inevitable compromises? Given the selective nature of the rewards, as they now stand, the probability of being able to so do is remote. The influence of the non-gay community, as well as the attitudes and beliefs of the majority of gays who do not belong to any gay organizations, hampers success. PMID- 10807030 TI - Part 3: A community divided. AB - The task of those who seek to encourage and offer social support has become more difficult as the majority of social institutions, and the state have established, over time, stronger and more pervasive modes of communication. The intricacies of gay identity have been articulated largely by forces outside of the gay movement, with the inevitable result that GSOs and ASOs have occupied less space in the consciousness of gay men. Additionally, I hypothesize that men who are HIV positive are engaging in fewer sexual contacts than men who are HIV-negative, and consequently lessening their attendance at venues where cruising is the main event. Financially and structurally, I shall demonstrate the disparity between GSOs and ASOs, and suggest that there is a natural intersection wherein the two SMOs could, and ought to cooperate, especially in the areas of fund-raising, joint program development, recruitment and political lobbying. However, their ideological bases appear to be sufficiently different to preclude such affiliations. These disparate ideologies are amplified by the mass media, and are consequently internalized by the members of the gay community. In the longer term, the divisiveness that manifests itself in the proliferation of numerous collectivities within the gay movement will contribute to the further isolation of gays from each other, and thwart any future attempts at coalition building, which could obviate the continued existence of a gay movement. Some writers suggest that the gay movement is going through a phase in an inevitable process of paradigm shifting, and in the end, the community will come back together stronger and more unified than it was previously. However, if this is a phase it is clear that the gay movement is in the 'dark before the dawn' initial phase of this paradigmatic shift, and subsequent phases are by no means guaranteed. As AIDS spreads beyond marginalized groups, and infiltrates the social majority, it is possible that much of the discrimination that has positioned gay men as 'other' will be abandoned in favour of a more enlightened, pluralistic conviction of the humanity of gays as full-fledged members of a mosaic-type community structure. On the other hand, it is also possible that as AIDS spreads into mainstream Western communities, gays will be further vilified and scapegoated as the perpetrators of this deadly disease. Preventative action is required to offset this possibility-preventative action can be affected by building a strong and unified gay community ready to withstand the onslaught of the mainstream enmity. This action would frame AIDS and gayness such that this type of situation would not come about. What is required is a new mode of co-operation among ASOs and GSOs, a model which firstly puts the gay house in order, and is then suitably structured to be more inclusive of all gay men's needs, and positioned to assist in the second wave of HIV infection-the general public. PMID- 10807031 TI - From Terra incognita to Terra firma: the logbook of the voyage of gay men's community into the Israeli public sphere. AB - This article charts the winding and tumultuous course Israeli gay men have taken in their struggles to claim a visible and audible place within the Israeli public sphere. Whereas for the greater part of history, Jewish gay men were symbolically annihilated by various social institutions, for the past decade they have been active as agents of social change. The paper's objectives are to offer an account of the developments that enabled such a transformation, to review socio-political strategies in an arena not yet discussed in academic literature, and to examine the roles mass media play in these processes. The article is composed of four chapters: literature review, detailed descriptions of the erstwhile and present legal, social, and cultural status of gay men in Israel, and a brief discussion of Orthodox-Jews' reactions. As this is a preliminary, and the first of its kind, study, it combines analyses of a diversified melange of sources. The author deliberately employs an eccletic methodological framework, nonetheless with an emphasis on newspapers as viable texts. PMID- 10807032 TI - Molecular modeling of 2-alkyloxy- and 2-aralkyloxy-adenosine A1- and A2-agonists. AB - The C2-region of adenosine A1- and A2-receptors by a molecular modeling technique has been extended and applied to a series of 2-substituted adenosines reported by Olsson, et al. The similarity and dissimilarity of the structure maps obtained by molecular modeling have been used as a basis for the mapping of the analysed receptor domain. The proposed model of the C2-region of the A1-receptor consists of a narrow and sterically limited area that interacts well electrostatically with small and electron rich moieties. Olsson's provisional model of the C2 region of the A2-receptor has been extended with two subsites, as well as with a forbidden area near the C2-position of the purine ring. The conformational analysis performed in the study does not support the hypothesis of Olsson et al. that adenosine C2 substituents may partly occupy the same receptor domain as the N6 substituents of the A1-receptor. The occupation of the cycloalkyl subsite increases the receptor selectivity while the occupation of the other subsite by aryl rings, fixed at a parallel position to the purine system, highly enhances the receptor affinity. PMID- 10807033 TI - Synthesis and 5-HT3 receptor affinity of new quinolinecarboxylic acid derivatives. AB - A series of quinolinecarboxylic acid amides and an ester with a quinuclidine moiety were synthesized and their in vitro affinities at 5-HT3, 5-HT4, and D2 receptors evaluated by radioligand binding assays. Highest affinity at 5-HT3 receptor corresponded to derivative 5 with Ki = 9.9 nM and with selectivity over 5-HT4 and D2 receptors. Compounds displayed moderate 5-HT3 antagonist activity (ED50 = 10.5-21.5 microg/kg i.v.). The obtained data suggest that the 5-HT3 receptor sites can accommodate the acyl group of the 2-quinoline derivatives. The results indicate the existence of an optimal distance between the lone electron pair of the quinoline nitrogen atom and the azabicyclic nitrogen atom, and a no pharmacophoric pocket in the 5-HT3 receptor which would hold the fragment at the position 4 of the quinoline ring. PMID- 10807034 TI - Development of a novel class of monocyclic and bicyclic alkyl amides that exhibit CB1 and CB2 cannabinoid receptor affinity and receptor activation. AB - CB1 and CB2 cannabinoid receptors can be activated by several different classes of agonists, including cannabinoids such as delta9-tetrahydrocannabinol and 9-nor 9beta-hydroxyhexahydrocannabinol, and eicosanoids such as arachidonylethanolamide. Structure-activity relationship studies have identified potential pharmacophoric elements for binding to cannabinoid receptors by both cannabinoids and eicosanoids. Molecular models have hypothesized conformational, spatial, and pharmacophoric distance requirements based upon radioligand binding data whereby overlap of pharmacophoric elements of the two classes disclose a low energy conformation of arachidonylethanolamide that can occupy the same receptor space as cannabinoid ligands. To test this model, we have developed a novel class of monocyclic and bicyclic alkyl amide cannabinoid receptor ligands. Further, we predicted a spatial conformation for these compounds in a molecular model based on the pharmacophoric and structural requirements for binding to the CB1 cannabinoid receptor. PMID- 10807035 TI - New antiherpetic 1,3-phenylene derivatives, inhibitors of the interaction of the HSV-1 origin binding protein (OBP) with DNA. AB - The synthesis of new 1,3-phenylene derivatives and their preliminary evaluation as antivirals (Herpes simplex 1, HSV-1) whose antiherpetic activity can be related with the inhibition of the interaction of the origin binding protein (OBP) with the DNA are presented. The new compounds are adjusted to a previously defined common structural model, consisting of a central aromatic system, which presents two side chains of different lengths in relative position 1, 3; these chains are made up of atomic groups characterized by the alternation of positive and negative centers, situating differently substituted rings, preferably aromatic, at the ends of both chains. Some of these derivatives, such as N,N''-(4 methoxy-1,3-phenylene)bis[N'-(4-nitrophenyl)urea] (2c) or (1,3-phenylene)bis[N-(p tolyl)aminosulfonyl] (11b), show antiherpetic activity related to the proposed mechanism. PMID- 10807037 TI - The prediction of partitioning coefficients for chemicals causing environmental concern. AB - Various types of partition coefficients have been used to facilitate the prediction of the concentration of pollutants in different phases in the environment. Many thousands of chemicals may exist in our environment which makes prediction work difficult or impossible due to a deficiency of knowledge of those unfamiliar compounds. In this study, the correlation between an octanol-water partition coefficient (Kow), water solubility (S) and a normalized soil/sediment partition coefficient (Koc) was investigated though the examination of 148 model chemicals. These model chemicals were classified into five major categories for easier adoption in future use. They are aliphatic compounds, aromatic compounds, pesticides, herbicides and polyaromatic hydrocarbons (PAH). Linear models are developed to correlate these partition coefficients in each category. The prediction of unfamiliar chemicals in the same category becomes possible if the fundamental properties of these chemicals (such as solubility) are previously known. PMID- 10807036 TI - Structure-activity relationship at the leucine side chain in a series of N,N dialkyldipeptidyl-amines as N-type calcium channel blockers. AB - Exploration of the SAR around the leucine side chain in a series of N,N dialkyldipeptidylamines with potent functional activity at N-type VSCC is presented. A novel analog is disclosed which possesses improved aqueous solubility, in vivo activity in an audiogenic seizure model, and reversible blockade in electrophysiological assays. PMID- 10807038 TI - Interspecific comparison of cadmium and zinc contamination in the organs of four fish species along a polymetallic pollution gradient (Lot River, France). AB - The impact of cadmium (Cd) and zinc (Zn) discharges related to an old zinc ore treatment facility in the Lot River (France) was investigated in four fish species (the chub: Leusciscus cephalus, the roach: Rutilus rutilus, the perch: Perca fluviatilis and the bream: Abramis brama). The organisms were sampled in four stations along the polymetallic contamination gradient. Cd and Zn analysis were carried out in five organs (gills, posterior intestine, liver, kidneys and skeletal muscle) in order to highlight the potential pathways of uptake, storage and elimination of metals. The results indicate a very strong Cd contamination in fish collected downstream from the metal source. The kidneys have the highest cadmium concentrations, but the gills and the intestine, as exchange organs, present the largest variations between the stations in close relation with the contamination gradient. Cd concentrations measured in the liver vary only slightly among the sampling stations. Unlike the trends observed for Cd, Zn levels in fish populations are strongly regulated and do not follow ambient Zn concentrations. The concentrations measured vary also according to fish species, for both Cd and Zn. This study shows that the trophic habits can explain the interspecific differences in Cd bioaccumulation. Zn levels observed for each species in non-contaminated populations also help to understand metal bioaccumulation patterns in polluted sites, suggesting that the determinism of interspecific differences is constitutive. PMID- 10807039 TI - Sedimentation rates and pollution history of the eastern Mediterranean Sea: Syrian coast. AB - Sedimentation rates along the Syrian coast were determined by the 210Pb dating method to a total of nine cores taken in 1993; rates were found to vary between 0.11 and 0.87 cm year(-1). These rates were used to provide historical records of the total organic content, major elements such as Al, Ca, Fe, K, Mg, Mn, Na, Ba and Br and some trace metals (As, Cr, Pb, Zn and Cu) concentrations along the Syrian coast. High total organic content up to 52% was found near Banias City; the Banise oil refinery and oil export activities were identified to be the pollution source of that area. Down core concentration profiles of trace metals show an increase in the concentrations of As and Cr in the upper layers for some of the collected cores. The highest values measured for As and Cr were 186 and 380 ppm, respectively, which are much higher than the natural levels. These high levels may be due to discharges of chemical wastes either from ships or coastal factories. However, the data can be used as a base background level for a future marine monitoring program of the Syrian coast. PMID- 10807040 TI - Total mercury in human renal cortex, liver, cerebellum and hair. AB - The aim of this study was: (a) to estimate the concentration of total mercury in the renal cortex, liver, cerebellum and hair of 46 persons who died suddenly in the Gdansk region, northern Poland, between the ages of 17 and 90; and (b) to assess whether a correlation occurs between mercury content in the investigated biological media. The mean concentrations of mercury in the human hair, renal cortex, liver and cerebellum were: 378 +/- 315.4 ng/g; 68.6 +/- 92.3 ng/g; 29.9 +/- 22 ng/g and 5.3 +/- 6.9 ng/g respectively. Positive correlations were found between mercury levels in: cerebellum and liver (r = 0.873), cerebellum and hair (r = 0.853), cerebellum and renal cortex (r = 0.578), hair and liver (r = 0.771), hair and renal cortex (r = 0.478) liver and renal cortex (r = 0.66). The geometric mean levels of mercury in the renal cortex, liver, cerebellum and hair in the residents of the Gdansk region are 15-19 times lower than in the population of Tokyo and its environs [Suzuki T, Hongo T, Yoshinaga J et al. The hair-organ relationship in mercury concentration in contemporary Japanese. Arch Environ Health 1993;44:361-365]. PMID- 10807041 TI - Effect of lead concentration on the level of glutathione, glutathione S transferase, reductase and peroxidase in human blood. AB - Incubation of human whole blood for 24 h at 37 degrees C in the presence of 100 400 microg/dl lead chloride or lead acetate caused a concentration-dependent decrease in the level of reduced glutathione up to 40%. Similarly, the activities of glutathione reductase, glutathione peroxidase and glutathione S-transferase were decreased up to 25%, 50%, and 19%, respectively. Moreover, 100 microg/dl lead chloride or lead acetate slowed the process of glutathione regeneration, and delayed the time for complete regeneration from 20 to 40 min. When glutathione S transferase was purified by affinity chromatography on Sepharose-linked glutathione, incubated with lead chloride or lead acetate, a concentration dependent inhibition of the enzymatic activity was observed reaching 50% inhibition at a lead salt concentration of 6000 microg/dl. PMID- 10807042 TI - Improving the reproducibility of the MCF-7 cell proliferation assay for the detection of xenoestrogens. AB - The MCF-7 cell proliferation assay is potentially a simple and highly reproducible tool for the identification of estrogenic compounds. However, its widespread use has been complicated by the lack of a standardised protocol, resulting in considerable inter-laboratory variability. We have explored the sources of variability both in relation to cell lines and test regimens and report on optimised procedures for the identification of estrogenic agents. Two supposedly identical MCF-7 parent cell lines (designated UCL and SOP), and the BUS subline were cultured according to an existing protocol, and responses to 17 estradiol (E2) assessed. Despite yielding almost identical EC50 values, the proliferative response varied widely between cell lines from 0.98-fold over controls (UCL) to 8.9-fold (BUS) indicating major differences between them. The underlying causes may be genetic, and to assess this we used comparative genomic hybridisation (CGH), a technique which allows the detection of DNA sequence copy number changes on a genome-wide scale. Although numerous similarities existed between the different cell lines, the least oestrogen-responsive line (MCF-7/UCL) exhibited the greatest number of cytogenetic changes, many of which were not seen in MCF-7/SOP cells. We suggest that care must be taken, therefore, when choosing a cell line for MCF-7 cell-based experiments. Selecting the MCF-7/SOP line for further work, we carried out a thorough and systematic optimisation of the MCF-7 cell proliferation assay, finding that a 72-h period in oestrogen-free medium before treatment strongly influenced the cells response to E2. With 1 nM E2, proliferation increased from 1.5-fold to 6.5-fold relative to vehicle-treated controls, a response similar to that seen with MCF-7/BUS cells in the E-SCREEN protocol devised by Soto et al. With parent MCF-7 cells, other laboratories have reported only 4.5-fold increases as maximal. Here we present evidence that the choice of cell line and culture conditions are crucial in determining test outcomes, and once chosen and adhered to the assay yields reproducible results. PMID- 10807043 TI - The use of epiphyton and epilithon data as a base for calculating ecological indices in monitoring of eutrophication in lakes in central Sweden. AB - Eutrophication was monitored with the aid of one similarity and seven diversity indices in the lakes of different trophic levels (eutrophic, mesotrophic and oligotrophic) in central Sweden. The ecological indices were applied separately to epiphyton and epilithon communities. Epiphyton turned out to be inappropriate for assessing eutrophication in the lakes studied. On the other hand, Hurlbert's, Simpson's and the similarity indices turned out to be promising environmental tools when applied to the data of epilithon. PMID- 10807044 TI - The NFkappaBb/IkappaB system in acute inflammation. AB - The transcription factor NFkappaB is a primary regulator of a wide variety of proinflammatory mediators. Under normal conditions, NFkappaB is retained in the cytoplasm bound to inhibitory proteins of the IkappaB family. Following cell activation, a number of signal transduction pathways lead to degradation of IkappaB proteins which results in nuclear translocation of NFkappaB and the ensuing transcriptional activation of proinflammatory genes. Several endogenous regulatory mediators, which function to prevent uncontrolled inflammation, exert their effects by blocking the activation of NFkappaB. Thus, NFkappaB appears to be at the heart of the acute inflammatory response. The present review discusses the role of NFkappaB in the induction and propagation of the acute inflammatory response as well as the regulation and resolution of this process. PMID- 10807045 TI - Chemokines, G proteins and natural killer cells. AB - Natural killer (NK) cells are anti-tumor and anti-viral effector cells. Members of C, CC, CXC and CX3C chemokines induce the chemotaxis and enhance the cytotoxicity of NK cells, suggesting that these cells express receptors for chemokines. The ability of members of chemokines to inhibit the replication of HIV-1 strains, combined with the ability of the same chemokines to activate the anti-viral NK cells, provide compelling evidence for the role of NK cells in eradicating HIV-1 infection. In addition, chemokines induce various intracellular signaling pathways in NK cells, which include activation of the heterotrimeric, and perhaps the small guanine nucleotide binding (G) proteins, as well as the mobilization of intracellular calcium, among other activities. Further, chemokines induce the phosphorylation of chemokine receptors through the recruitment of G protein-coupled receptor kinases (GRKs) resulting in the desensitization and turning off the signals. In this review, I will update the knowledge of the effect of chemokines on NK cell motility and the signal transduction pathways induced by chemokines in these cells. PMID- 10807046 TI - Immune privilege or inflammation? The paradoxical effects of Fas ligand. AB - Fas ligand (FasL) induces apoptosis of cells, including activated lymphocytes, expressing its cognate receptor, Fas (CD95/APO-1). FasL precludes inflammatory reactions from immune privileged sites by triggering Fas-mediated apoptosis of infiltrating proinflammatory cells. Aberrant expression of FasL by cancers inhibits antitumor immune responses. The ability of FasL to impair immune responses may hold therapeutic promise as a means of protecting tissue transplants from immunological rejection. Paradoxically, FasL exhibits proinflammatory activity independent of its ability to mediate immune privilege. FasL has been shown to recruit and activate neutrophils, although the factors that determine whether FasL is pro- or anti-inflammatory are only beginning to emerge. FasL appears to contribute to cell death in Fas-sensitive endorgan cells during inflammation. Blocking of Fas-mediated endorgan apoptosis or enhancing Fas mediated apoptosis of inflammatory cells represent potential targets for future antiinflammatory therapies. PMID- 10807047 TI - Immunotherapy of inflammatory bowel diseases: current concepts and future perspectives. AB - The etiology and the pathogenesis of inflammatory bowel diseases (IBD), e.g. Crohn's disease and ulcerative colitis are still not completely understood. However, there is growing evidence that an alteration of the mucosal immune system towards luminal antigens in a genetically susceptible host plays a key role in the pathogenesis of IBD. In particular, cytokines produced by intestinal epithelial cells, lamina propria macrophages and CD4+ T cells appear to contribute to the initiation and perpetuation of intestinal inflammation in IBD. This review focuses on the role of the mucosal immune system in the pathogenesis of IBD and potential novel immunotherapeutic strategies for chronic intestinal inflammation. Such strategies include recombinant antiinflammatory cytokines, neutralizing antibodies or fusion proteins, antisense oligonucleotides and adenoviral gene transfer. PMID- 10807048 TI - Tumor vaccines for the management of prostate cancer. AB - Prostate cancer is a significant health problem and one of the leading causes of cancer-related death among men. Given the typically long natural history of the disease, there is considerable interest in developing new therapies to treat or prevent metastatic disease, and cancer vaccines are a particularly attractive immune-based approach. Early clinical studies using non-specific immunomodulatory treatments have met with limited success, but also suggest that improved immunologic approaches might be useful in treating human prostate cancer. Over the last decade, the identification of immune cells responsible for actual destruction of prostate tissue and advances in immunologic and molecular techniques have led to a variety of vaccination approaches that are currently being evaluated in human clinical trials. The present article discusses the rationale in animal models for particular immunization strategies and describes the vaccines currently being used in patients with prostate cancer. The ongoing identification of tumor antigens and proteins involved in prostate cancer progression and the development of better immunologic animal models suggest a hopeful future for the design of effective prostate cancer vaccines. PMID- 10807049 TI - The genes of interferons and interferon-related factors: localization and relationships with chromosome aberrations in cancer. AB - The paper presents a review of data on the localization of interferons (IFNs) and IFN system genes and their relationship with human diseases, mainly cancer. Genes of interferon system proteins are located at the sites of breakpoints of the structural chromosome aberrations in cancer. Thus, any of them are rearranged or translocated in various tumor types. As the activity of these genes plays a role in cancer development, their rearrangements may be one of the crucial points in the pathogenesis of some cancer types. Besides, they also take part in organism immunity against viral infections. Transfection experiments with IFN system genes have proved the influence of these genes on cancer behavior and may serve as a basis for clinical gene therapy. IFN-alpha and IFN-beta genes are located at 9p21 22, the site of frequent homozygotic deletions in cancer. Their loss sensitizes cells to the growth inhibitory actions of exogenous IFNs. The IFN-gamma gene, a representative of class II genes, is located at 12q24.1. Transfection of class II IFNs genes to cancer cell lines causes cell proliferation arrest and augments the expression of HLA antigens, which may be clinically useful in stimulating the immune destruction of tumor cells. The interferon regulatory factor 1 (IRF-1) gene is located at 5q31, the site of common deletions in myelodysplastic syndromes (MDS) and secondary leukemias. The loss of heterozygosity of this gene was found in MDS, which proves that IRF-1 may be a tumor suppressor. A transfection of its gene causes neoplastic transformation arrest. The double stranded RNA-activated protein kinase (PKR) gene is located at 2p21-22, a region which is frequently rearranged in leukemia. Transfection of a wild type PKR gene reverses neoplastic transformation caused by transfection of a mutated PKR gene, proving that PKR acts as a dominant negative cancer suppressor. PMID- 10807050 TI - Interleukin 6, tumor necrosis factor alpha and their soluble receptors in the blood serum of patients with denture stomatitis and fungal infection. AB - Determinations of the blood serum levels of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and their soluble receptors (sIL-6R, sTNFR) in denture stomatitis patients (DS) were performed. Serum levels of interleukins and their soluble receptors were measured using the ELISA method. In all examined patients mycological diagnostics were conducted using API 20C AUX stripe tests and an automatic ATB machine. Results were compared with those of healthy denture wearers (D), and controls (C). In DS patients, yeasts were isolated in 90.9%, in D in 66.7% of cases. The most often isolated species in both groups was Candida albicans. Mean concentrations of IL-6 and TNF-alpha were statistically significantly higher in DS and D groups compared to controls. Mean concentrations of sIL-6R were similar in all groups; however, concentrations of sTNFR in both DS and D groups were significantly lower compared to controls. There were no correlations found between values of IL-6 and TNF-alpha nor between examined interleukins and their soluble receptors. PMID- 10807051 TI - Histamine receptor expression on peripheral blood lymphocytes is influenced by specific and nonspecific activation. AB - Histamine is a physiological mediator which exerts both effector and regulatory functions through its receptors on various cells. The aim of the study was to investigate changes in histamine receptor expression on peripheral blood lymphocytes affected by stimulation with both specific and nonspecific stimuli. Lymphocytes were obtained from both healthy and allergic subjects. Cells were incubated with various allergens (mixed grass pollen, Lolium perenne, Dermatophagoides pteronyssinus 1, bee venom, phospholipase A2) and nonspecific (fMLP, PMA/ionomycin, LPS) stimuli. The percentage of histamine-binding cells was determined with a fluorescence microscope after incubation with histamine fluorescein. In control subjects histamine binding after stimulation with allergens was not significantly changed. In contrast, in allergic subjects stimulation with specific allergens resulted in significantly increased histamine binding. Nonspecific stimulation caused increased histamine binding to lymphocytes in both allergic subjects and healthy controls. We conclude that specific and nonspecific activation of lymphocytes is associated with increased expression of histamine receptors. PMID- 10807052 TI - T cell depleted haploidentical bone marrow transplantation for the treatment of children with severe combined immunodeficiency. AB - Severe combined immunodeficiency (SCID) is fatal in early childhood if unrecognized and if not treated. The aim was to determine the efficacy of T cell depleted bone marrow transplantation (TCD BMT) in the treatment of children with SCID. Eleven children diagnosed with SCID received histocompatible related donor bone marrow transplantation--HRD BMT (group I). Thirty seven children diagnosed with SCID who did not have histocompatible donors were treated with TCD haploidentical parental bone marrow transplantation (BMT) (group II). TCD was performed by in vitro soybean lectin agglutination followed by E-rosette depletion. Patients were longitudinally assessed for the presence and function of T and B lymphocytes. In group I all children survived. The mean age of children in this group at the time of HRD BMT was 15.4 months. All surviving patients normalized their specific T cell function. Two out of 11 require treatment with intravenous immunoglobulin i.v. Ig. In group II 17 out of 37 (46%) children survived. At the time of TCD BMT the mean age of survivors was 7.5 months, vs. 11.4 months in patients who died. Death was caused most commonly by opportunistic infections, Epstein-Barr virus induced lymphoproliferative disease (EBV-LPD), and graft versus host disease (GvHD). Seventeen out of 17 surviving patients recovered normal numbers of CD3+ cells and antigen specific T cell function. Five out of 17 never recovered their B cell function and require i.v. Ig injections. Early diagnosis, prevention or treatment of opportunistic infections, and enhancement of immune recovery will be necessary to improve survival in patients with SCID treated with TCD BMT. PMID- 10807053 TI - The contribution of endotoxins present in the respiratory tract to overproduction of nitric oxide associated with impaired interleukin 6 release in bronchoalveolar leukocytes from lung cancer patients. AB - The purpose of the study was to assess the relation between the levels of endotoxins circulating in airways of patients with lung cancer and the ability of bronchoalveolar lavage (BAL) leukocytes for ex vivo release of nitric oxide (NO) and interleukin 6 (IL-6) and for in vitro lipopolysaccharide (LPS)-induced production of the mediators. Leukocytes isolated from the BAL of 11 patients and from 5 healthy individuals were cultured in the absence or presence of LPS(E. coli). The levels of endotoxins in the BAL fluids (BALF) and the amounts of NO released ex vivo from unstimulated cells from the patients were highly (p = 0.0025) elevated in comparison with those from healthy individuals. The release of NO was significantly correlated (R(S) = 0.638, p = 0.047) with the levels of endotoxins in BALF. In contrast, production of IL-6 remained very low and a negative correlation (R(S) = -0.623, p = 0.0542) was observed between the amounts of NO and IL-6. It was also found that, in response to LPS, bronchoalveolar leukocytes from patients with lung cancer express a reduced capacity for in vitro production of NO and IL-6. Our data suggest that, in patients with lung cancer, the activation of BAL cells by endotoxins circulating in the airways may contribute, at least in part, to overproduction of spontaneous NO and, subsequently, the NO may reduce IL-6 production. Moreover, the exposure in vivo of the BAL cells to LPS renders them unable to respond to the second signals. PMID- 10807054 TI - RM-11, a new izoxasole derivative, is a potent stimulator of the humoral and cellular immune responses in mice. AB - In this report we describe immunostimulatory properties of RM-11 in several in vivo and in vitro tests in the murine model. We found that RM-11 significantly stimulated the humoral immune response to sheep erythrocytes (SRBC) when given intraperitoneally (i.p.) at doses of 10 and 100 microg or per os (doses of 20 and 200 microg) 3 h before immunization. The compound was also stimulatory with regard to generation of delayed type hypersensitivity (DTH) to SRBC when given i.p. or per os (doses of 10, 100 and 500 microg/mouse). The described immunostimulatory activities of RM-11 were higher compared to that of the reference drug, levamisole. RM-11 stimulated, in addition, concanavalin A (ConA) induced splenocyte proliferation. Lastly, we showed that RM-11 was not toxic when given to mice per os at doses 250 mg/kg body weight. Taken together, RM-11 appeared to be a universal stimulator of the immune response in mice. Lack of toxicity and the ability to stimulate the immune response, when administered per os, predispose the compound for further preclinical studies. PMID- 10807055 TI - On handicapping thyroidology. PMID- 10807056 TI - The localization of thyroid hormone receptor mRNAs in human bone. AB - Thyroid hormones have well-documented effects on the skeleton although the mechanism of their action on bone is poorly understood. We have recently reported the presence of different thyroid hormone receptor isoforms in human bone. However, there is evidence to suggest that the expression of thyroid hormone receptor (TR) protein may not necessarily correlate with its mRNA. In this study, we used specific digoxigenin-labeled ribo probes to investigate the expression of TRalpha1, variant TRalpha2, TRbeta1, and in particular TRbeta2 mRNA in human osteophytic bone and osteoclastoma tissue in situ. The number of positive cells was expressed as the percentage of the total number of cells of the same phenotype. In osteophytes, at sites of endochondral ossification, TRalpha1, variant TRalpha2, TRbeta1, and TRbeta2 mRNA were widely distributed in undifferentiated, proliferating, mature and hypertrophic chondrocytes. At sites of bone remodeling, TRalpha1 mRNA was expressed in the majority (> 90%) of osteoblasts. TRbeta1 and the variant TR-alpha2 mRNA were moderately expressed in approximately 75% of cells with only a few osteoblasts (< 25%) expressing TRbeta2 mRNA. All the TR transcripts were highly expressed in multinucleated osteoclasts in osteoclastoma tissue. The distribution of TR mRNAs was similar to TR receptor protein expression (as we have previously reported) in both osteophytic bone and osteoclastoma tissue except TRalpha1 mRNA that was highly expressed in osteoclasts and in undifferentiated, proliferating, mature, and hypertrophic chondrocytes in contrast to its receptor protein expression. This study highlights the importance of studying both TR mRNA and receptor proteins in triiodothyronine (T3) responsive tissues. This is also the first demonstration of the presence of TRbeta2 mRNA in bone. The role of TRbeta2 in mediating the actions of thyroid hormones in bone is not known and requires further investigation. PMID- 10807057 TI - Role of insulin and serum on thyrotropin regulation of thyroid transcription factor-1 and pax-8 genes expression in FRTL-5 thyroid cells. AB - Thyrotropin (TSH), via its cyclic adenosine monophosphate (cAMP) signal, decreases thyrotropin receptor (TSHR) gene expression in FRTL-5 thyroid cells, whereas it increases expression of the thyroglobulin (Tg) gene. Despite the opposite effects of TSH on TSHR and Tg expression, both genes are positively controlled by thyroid transcription factor-1 (TTF-1) and evidence has accumulated that TSH can decrease TTF-1 mRNA levels. In this report, we further characterize the action of TSH on TTF-1 in order to understand its different activities on the TSHR and Tg genes better. The effect of TSH on the TSHR requires the presence of insulin and serum and we show here that also both factors are necessary for the TSH effect to decrease TTF-1 mRNA levels. The decrease is paralleled by a downregulation of TTF-1 protein levels as well as by a decrease in TTF-1/DNA complex when the TTF-1 site of the TSHR promoter was used as probe. Again, the decrease requires insulin and serum. The TSH downregulation of TTF-1 mRNA levels is due to a decrease in its transcription rate. Using a luciferase-linked chimera construct spanning 5.18 kb of the TTF-1 5'-flanking region, we show that TSH decreases TTF-1 promoter activity and that this effect depends on insulin and serum. These data contrast with the action of TSH on Tg and Pax-8 gene expression. TSH increases Pax-8 mRNA levels and the increase is evident whether insulin and serum are present or not. Moreover, this increase is paralleled by an increase in Pax-8 protein binding to an oligonucleotide derived from the C site of the Tg promoter, which can bind both TTF-1 and Pax-8. The present data thus show that TTF-1 gene expression is interdependently regulated by TSH and serum growth factors including insulin. They also show this interdependent-regulation is not duplicated in the case of Pax-8. We suggest that these differences may contribute to the distinct ability of TSH to regulate TSHR versus Tg gene expression in FRLT-5 thyroid cells. PMID- 10807058 TI - Mitogenic effect of lithium in FRTL-5 cells can be reversed by blocking de novo cholesterol synthesis and subsequent signal transduction. AB - Lithium therapy is the therapeutic mainstay for bipolar disorder and has been associated in the thyroid with euthymic goiter, hyper and hypothyroidism as well as thyroid autoimmune disease. The FRTL-5 cell line is a well known model of thyroid cell physiology, where lithium has been shown to increase 3H-thymidine uptake at concentrations of 2 mM. This mitogenic effect was not associated with adenylate cyclase as measured by cyclic adenosine monophosphate (cAMP) production. The de novo synthesis of cholesterol is an important signal transduction pathway in FRTL-5 cells, where newly synthesized Rho GTPase is geranylgeranylated, enabling membrane localization of the G-protein and subsequent G1 to S-phase transition, resulting from extracellular stimulation. Here we confirm lithium mitogenicity at therapeutically relevant concentrations (1 mM) and demonstrate a lithium-associated accumulation of FRTL-5 cells in S phase of the cell cycle. These effects could be abolished by Pravastatin, a potent inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA), the rate limiting enzyme in the formation of intermediates (de novo cholesterol synthesis) required for G-protein prenylation. Pravastatin, similar to lithium, showed no effect on cAMP production either under basal or thyroid stimulating hormone (TSH) stimulated conditions indicating that de novo cholesterol synthesis is not involved with adenylate cyclase. The inhibitory effect of pravastatin could be overcome by reinitiating de novo cholesterol synthesis. This was achieved by the addition of the cell permeable, first metabolite (mevalonate) after HMG-CoA, which allowed the cycle to continue, leading eventually to protein prenylation, despite the presence of Pravastatin. These novel findings demonstrate lithium involvement in de novo cholesterol synthesis and G-protein prenylation, an important signal transduction pathway in FRTL-5 cells. PMID- 10807059 TI - Gene therapy of established medullary thyroid carcinoma with herpes simplex viral thymidine kinase in a rat tumor model: relationship of bystander effect and antitumor efficacy. AB - Bystander effect (BSE) refers to killing of cells adjacent to a cell engineered to express a killing gene segment. BSE is considered an important aspect of suicide gene therapy with thymidine kinase. We evaluated the BSE of adenovirus expressing herpes simplex thymidine kinase (AdCMVtk) in rat medullary thyroid carcinomas (rMTC) and three rat thyroid epithelial cancer cell lines using an in vitro BSE assay. In the assay, different proportions of infected and uninfected cells are mixed. Only the proportion of directly infected cells was inhibited in the proliferation assay using rMTC cells. This indicates that there is little BSE in this cell line. One rat thyroid epithelial cancer cell line (RTC-R2) has a high BSE, with BSE index (BSEi) of 7. In the proliferation assay a greater proportion of cells was inhibited than those directly infected. BSE was also evaluated during in vivo tumor growth by subcutaneous injection of mixtures of AdCMVtk infected and uninfected cells. Ganciclovir (GCV) treatment of tumors developing from a 1:1 mixture of infected to uninfected rMTC cells failed to inhibit their growth. In contrast, GCV treatment of a 2:8 mixture of infected to uninfected RTC-R2 cells completely inhibit tumor development, indicating a high BSE. BSE is related to in vivo antitumor efficacy when replication-defective adenovirus AdCMVtk is directly injected into rMTC tumors. After treatment with 100 mg/kg per day of GCV, a growth-retardation effect was observed in small tumors (<100 mm3), but there was little antitumor activity in large tumors (>100 mm3). Our results indicate that there is a good correlation between this in vitro BSE assay and in vivo treatment efficacy. Not all kinds of tumors are suitable for thymidine kinase (TK)/GCV gene therapy because some lack BSE. Methods to improve BSE and/or transduction efficiency are needed in order to obtain an effective therapeutic result. It will be appropriate to test the BSE in human tumor cells before performing clinical trials with current adenoviral vectors expressing TK. PMID- 10807060 TI - Thyroid iodine transport. PMID- 10807061 TI - Worm-eaten bones. PMID- 10807062 TI - Fractures in patients with primary idiopathic hypothyroidism. AB - AIM: To study fracture risk and risk factors for fractures in patients with primary idiopathic hypothyroidism (ICD 10: E03.9). DESIGN: Historical follow-up. MATERIAL AND METHODS: A self-administered questionnaire was issued to 628 patients with primary idiopathic levothyroxine-substituted hypothyroidism. A total of 412 (65.6%) responded and of these, 408 could be analyzed. The 408 respondents were age- (+/- 5 years) and gender-matched with 408 normal controls randomly selected from the background population who responded to the same questionnaire. RESULTS: Overall fracture risk was increased in patients compared to controls (relative risk: RR = 1.6, 95% CI: 1.0-2.5). However, the increase was temporary and limited to the period within the first 2 years after the diagnosis of hypothyroidism (RR = 3.1, 95% CI: 1.4-7.0). Before the diagnosis and more than 2 years after the diagnosis, the fracture risk in patients did not deviate from that of the controls. The increase in fracture risk was only significant in the age group above 50 years (RR = 1.8, 95% CI: 1-3.2), and was limited to the forearms (RR = 3.0, 95% CI: 1.4-6.3 for the entire patient population). CONCLUSIONS: There was a temporary increase in fracture risk within the first 2 years after diagnosis of primary idiopathic hypothyroidism. The fracture risk was mainly increased in the age group above 50 years, and the increased risk was limited to the forearms. PMID- 10807063 TI - Fracture risk in patients treated for hyperthyroidism. AB - AIM: To study fracture risk and risk factors for fractures in patients with hyperthyroidism. DESIGN: Historical follow-up. MATERIAL AND METHODS: A total of 864 patients with diffuse toxic goiter (ICD 10: E05.0) or toxic nodular goiter (E05.2) were contacted through a self-administered questionnaire. Each respondent was compared to an age: (+/- 5 years) and gender-matched control from a random sample of the background population who responded to the same questionnaire. RESULTS: Among the patients 621 (72%) responded and of these 617 could be analyzed. Within the first 5 years before the diagnosis, the patients had the same fracture risk as the controls (RR = 1.2, 95% CI; 0.7-2.0). After the diagnosis, fracture risk was elevated among the patients (RR = 1.7, 95% CI: 1.2 2.3), especially in the age group 50 years or older (RR = 2.2, 95% CI: 1.5-3.3). Fracture risk was elevated for fractures of the spine (RR = 8.9, 95% CI: 1.6 48.4), and the forearms (RR = 3.1, 95% CI: 1.6-6.2), but not at other skeletal sites. Treatment with radioactive iodine alone was associated with an increased fracture risk (OR = 2.7, 95% CI: 1.2-6.0), a risk that was not present in patients who, in addition to radioactive iodine, also had received methimazole (RR = 1.5, 95% CI: 0.7-3.2). CONCLUSIONS: Our study demonstrated an increased fracture risk in hyperthyroidism, a fracture risk that was present with radioactive iodine treatment alone, but not in subjects that had received both radioactive iodine and methimazole or other types of antithyroid therapy. PMID- 10807064 TI - The expression of vascular endothelial growth factor and the type 1 vascular endothelial growth factor receptor correlate with the size of papillary thyroid carcinoma in children and young adults. AB - Vascular endothelial growth factor (VEGF) is essential for the growth of many solid tumors, but there are little data regarding VEGH in childhood thyroid cancers. We examined the relationships between VEGF, the type 1 VEGF receptor (FLT-1) and clinical outcome for a group of thyroid cancers in children and young adults. The expression of VEGF and FLT-1 were determined by immunohistochemistry using archival, paraffin-embedded thyroid tissue blocks and compared with the retrospective clinical outcome for each patient. The study included 67 children and young adults with papillary thyroid carcinoma (PTC, n = 42), follicular thyroid carcinoma (FTC, n = 8), benign lesions (n = 15), or controls (n = 2). VEGF expression was greater in PTC (mean intensity 2.23 +/- 0.25, p = 0.002) and FTC (2.8 +/- 0.73, p = 0.01) than benign lesions (1.0 +/- 0.27), and correlated with PTC size (r = 0.42, p = 0.008). FLT-1 expression was greater in PTC (mean intensity 2.8 +/- 0.17) than FTC (1.9 +/- 0.25, p = 0.015) and benign lesions (1.7 +/- 0.32, p = 0.002); and correlated with PTC size (r = 0.41, p = 0.01) as well as VEGF expression (r = 0.52, p = 0.002). Recurrent disease developed exclusively in patients with PTC which expressed VEGF (7/28, 95% CI 10.6%-44.2%). PTC that did not express VEGF (0/8, 95% CI = 0%-31.2%) did not recur; however, the difference was not statistically significant (p = 0.15). We conclude that the expression of VEGF and FLT-1 are directly correlated with the size of PTC in children and young adults. PMID- 10807065 TI - Unusual complication of total thyroidectomy: arteriovenous fistula. AB - We describe the case of a 29-year-old woman from the Chernobyl area, who moved to the Czech Republic 10 years after the Chernobyl nuclear disaster. She was evaluated for a large goiter. Fine-needle aspiration cytology indicated a diagnosis of a typical degenerated Hurthle cell adenoma. She underwent total thyroidectomy but a continual loud murmur was found 1 month after surgery. An arteriovenous (AV) fistula between the right thyrocervical trunk and the right jugular vein was proven angiographically. This fistula was successfully closed by artificial embolization performed by detachable Gianturco-Anderson-Wallace (GAW) coil. This is the first description of using such a procedure in order to close an AV fistula after thyroidectomy. PMID- 10807066 TI - Thyrotoxicosis in a patient with submandibular thyroid. AB - Ectopic thyroid glands generally appear in the midline due to abnormal median migration; their presence lateral to the midline is rare. We report the case of a 42-year-old female who presented with symptoms of thyrotoxicosis and an expanding submandibular swelling. Tc-99m-sodium pertechnetate scanning showed thyroid tissue in the left submandibular region, while no thyroid tissue was seen in the normal site. The patient was treated with 10 mCi of 131I and subsequently became euthyroid. Literature review revealed seven cases of lateral aberrant thyroid tissue. The theories to explain lateral aberrant thyroid are presented. PMID- 10807067 TI - Tracheal compression in a patient with substernal extension of a multinodular goiter. PMID- 10807068 TI - HLA-B40-, HLA-Cw3-, and HLA-DR5-associated susceptibility to amiodarone-induced thyroid dysfunction. PMID- 10807069 TI - Renal artery stenosis: the case for conservative management. PMID- 10807070 TI - Outcomes of atherosclerotic renal artery stenosis managed without revascularization. AB - OBJECTIVE: To determine how often patients with renal artery stenosis (RAS) managed without revascularization progress to accelerated hypertension and/or renal failure. PATIENTS AND METHODS: We examined the outcomes of 68 patients (mean +/- SEM age, 71.8 +/- 0.9 years) with high-grade (>70%) RAS identified between 1989 and 1993 who were treated without renal revascularization for at least 6 months after angiography. The time to last follow-up averaged 38.9 +/- 2.8 months. Other vascular beds were affected in 66 of the 68 patients. End points were revascularization, nephrectomy, dialysis, or death. RESULTS: The mean +/- SEM serum creatinine level rose from 1.4 +/- 0.1 to 2.0 +/- 0.2 mg/dL (P<.001). Mean +/- SEM blood pressure did not change (157 +/- 3/83+/-2 vs 155 +/- 3/79 +/- 2 mm Hg), but the need (mean +/- SEM) for medication increased from 1.6+/-0.1 to 1.9+/-0.1 drugs (P=.02). Four patients (5.8%) eventually underwent renal revascularization for refractory hypertension (1 patient), for progressive stenosis (1 patient), and during aortic reconstruction (2 patients). One additional patient underwent nephrectomy to improve blood pressure control. Five others (7.4%) developed end-stage renal disease (ESRD) for reasons other than progressive vascular disease, namely, diabetes (3 patients), atheroemboli (1 patient), and contrast toxicity without RAS progression (1 patient). In 1 further case, the reason for ESRD was unknown, and it may have been caused by vascular occlusion. During follow-up, 19 patients died of unrelated causes, including myocardial infarction and stroke. CONCLUSIONS: These data indicate that antihypertensive medication requirements increased and renal function deteriorated modestly in a subset of patients with atherosclerotic RAS managed initially without vascular intervention. Many achieved stable blood pressure for many years. Deterioration of renal function and mortality risk were greatest in patients with bilateral stenosis or stenosis to a solitary functioning kidney. These results reinforce the need for meticulous follow-up for disease progression but underscore the role of competing risks and high mortality from other cardiovascular diseases, which primarily determine the outcomes in patients with RAS and widespread atherosclerotic disease. PMID- 10807071 TI - The chronic ventilator-dependent unit: a lower-cost alternative to intensive care. AB - OBJECTIVE: To evaluate the fixed costs and patient outcomes of a specialty hospital unit for medically stable ventilator-dependent patients. The chronic ventilator-dependent unit (CVDU) was established to facilitate early dismissal from costly intensive care unit (ICU) hospitalization for patients requiring continued specialized care. PATIENTS AND METHODS: We carried out a cost analysis of the various ICUs that transferred patients to the CVDU by year from 1993 through 1998. In addition, direct and indirect costs for the CVDU were established by year for the same period. We then calculated the cost effect of transferring these patients for care from each high-cost ICU to the lower-cost CVDU. Ventilator weaning and mortality rates were also determined. RESULTS: During the 6 years of this study, $4,832,551 in patient care costs were saved by transferring care for 964 patients from ICUs to the CVDU. Ventilator weaning was successful in 64% of 549 patients, and mortality was 7% in the same patient group. CONCLUSIONS: Care in the CVDU yielded lower fixed costs per patient-day, and CVDU care was comparable to ICU hospitalization. PMID- 10807072 TI - Alpha1-antitrypsin deficiency and inflammatory bowel diseases. AB - OBJECTIVE: To evaluate a possible etiologic role of alpha1-antitrypsin deficiency (alpha1AD), most frequently caused by a Z allele mutation, in ulcerative colitis (UC) and Crohn disease (CD). PATIENTS AND METHODS: This retrospective study included 10 patients diagnosed with and/or treated for inflammatory bowel disease (IBD) between 1976 and 1997 and identified from the Mayo Clinic Medical Index System. All 10 patients had either alpha1AD and CD or alpha1AD and UC. The alpha1 antitrypsin (alpha1AT) types and levels were determined with isoelectric focusing testing. The allele types, representing genotypes, were designated PiZZ (or ZZ) for homozygotes and PiMZ (or MZ) for heterozygotes. RESULTS: Seven patients had UC: 4 were genotype ZZ and 3 MZ. Four of these 7 patients had emphysema, 2 had asthma, and 1 had chronic bronchitis. Five were cigarette smokers, but only 1 had smoked coincident with activity of her UC. Two of the UC patients never smoked, and 1 of these 2 had asthma. Three of the 10 patients had CD, 2 genotype ZZ and 1 MZ. Two of the 3 patients were long-term cigarette smokers, and both had emphysema. Nine of the 10 patients with UC and alpha1AD required surgery. CONCLUSIONS: The need for surgery in patients with UC and alpha1-AD may point to a unique phenotypic subgroup of patients with alpha1AD and severe UC. Further studies are required to substantiate the etiologic role of alpha1AD in IBD. Our observations, if confirmed by future studies, suggest that in patients with both IBD and chronic obstructive pulmonary disease, alpha1AD testing should be considered. PMID- 10807073 TI - Diet drug-related cardiac valve disease: the Mayo Clinic echocardiographic laboratory experience. AB - OBJECTIVE: To describe the prevalence of diet drug-related valvular disease among our referral population and the association of valvular disease with duration of exposure to fenfluramine and phentermine in combination and to dexfenfluramine alone. PATIENTS AND METHODS: In this retrospective review of clinical and echocardiographic data, charts of patients referred for treatment of toxic effects of diet drugs were reviewed, and telephone interviews were conducted. RESULTS: Between June and December 1997, 191 patients (164 women, 27 men; mean age, 47 years) were referred for possible diet drug-related valvular disease. Twenty-eight (28%) of the 99 asymptomatic patients and 40 (43%) of the 92 symptomatic patients had abnormal echocardiographic findings. Valvular lesions among the 68 patients with abnormal echocardiographic findings included mild (or greater) aortic regurgitation in 55 patients (81%), moderate (or greater) mitral regurgitation in 12 (18%), and moderate (or greater) tricuspid regurgitation in 7 (10%). The Food and Drug Administration case definition of diet drug-related valvulopathy was noted in 31 % of this referral population. Of patients with valvulopathy, mean duration of therapy with fenfluramine and phentermine in combination and dexfenfluramine alone was 9 months and 5 months, respectively. Duration of therapy was not associated with presence or absence of disease. Five patients had surgical intervention for severe valvulopathy: 3 had mitral valve repair, 1 had mitral valve replacement, and 1 had aortic valve replacement. Pulmonary hypertension (>40 mm Hg) was found in 24 patients (13%), and 17 (71 %) had pulmonary hypertension in association with valvulopathy. CONCLUSION: This study demonstrated a 31% (60/191) prevalence of valvulopathy in patients with a history of diet drug exposure who were referred for echocardiographic evaluation. The most common finding was mild aortic regurgitation. Twenty-eight percent of asymptomatic patients had abnormal echocardiographic findings. This study emphasizes the spectrum of diet drug-related cardiac disease and the potential for valvulopathy in asymptomatic patients. PMID- 10807074 TI - Differentiation of typhoid fever from fulminant hepatic failure in patients presenting with jaundice and encephalopathy. AB - OBJECTIVE: To determine the clinical and laboratory features that allow the early diagnosis of typhoid fever in patients who present with jaundice and encephalopathy. PATIENTS AND METHODS: This 12-month prospective study, conducted in Bangalore, India, between 1990 and 1991, evaluated the clinical and laboratory features of all patients (N=47) who presented with encephalopathy within 8 weeks of onset of jaundice. Ciprofloxacin and dexamethasone were used to treat 11 patients diagnosed on blood culture as having typhoid fever. The other 36 patients were presumed to have fulminant hepatic failure with a viral cause and were treated with supportive measures (bioartificial liver support and transplantation were not available). RESULTS: In patients with jaundice and encephalopathy, a liver span of greater than 9 cm on physical examination, thrombocytopenia, elevated alkaline phosphatase level, aspartate aminotransferase level greater than alanine aminotransferase level, and only mild prolongation of the prothrombin time suggested a diagnosis of typhoid fever. All 11 patients diagnosed as having typhoid fever had an excellent response to treatment with ciprofloxacin and dexamethasone with no mortality and with normalization of the liver test results in 2 weeks. On the other hand, 30 of the 36 patients with nontyphoid fulminant hepatic failure died. CONCLUSIONS: In patients presenting with jaundice and encephalopathy, physical examination and simple laboratory tests can help make an early diagnosis of typhoid fever. We believe that patients with a presumptive diagnosis of typhoid fever should be treated with ciprofloxacin and dexamethasone, even before the results of blood cultures are available. PMID- 10807075 TI - The shrinking lungs syndrome in systemic lupus erythematosus. AB - A comprehensive review of the literature on shrinking lungs syndrome (SLS) in systemic lupus erythematosus involved a MEDLINE search (1965-1997) of case reports and clinical series of patients with the diagnosis of SLS. A total of 49 well-documented cases of SLS were reviewed. Shrinking lungs syndrome is characterized by unexplained dyspnea, a restrictive pattern on pulmonary function test results, and an elevated hemidiaphragm. The cause of SLS remains controversial, with several authors attributing the disorder to diaphragmatic weakness and others suggesting that chest wall restriction accounts for the clinical syndrome. No definitive therapy exists. Corticosteroids have been reported to lessen symptoms and improve pulmonary function in some patients with SLS, but other methods of treatment have occasionally been found to be helpful. Clinical presentation, method of diagnosis, pathogenesis, and treatment modalities are summarized in this review. An uncommon complication of systemic lupus erythematosus, SLS causes significant morbidity and, occasionally, mortality. PMID- 10807076 TI - Henry Heimlich--Heimlich maneuver. PMID- 10807078 TI - Overview of skin cancer detection and prevention for the primary care physician. AB - The incidence of both melanoma and nonmelanoma skin cancer is increasing, and an accurate and timely diagnosis is important to reduce the morbidity and mortality associated with these malignancies. Clinical features and examination techniques are discussed, and the need to recognize important secondary tumor characteristics, which supplement the primary diagnosis, is reviewed. Recent evidence on etiologic factors and their importance are addressed to highlight the role of primary prevention. Pediatricians, family physicians, and all primary care physicians are encouraged to promote skin cancer awareness among children and their parents. PMID- 10807077 TI - Hereditary ataxias. AB - There are many causes of hereditary ataxia. These can be grouped into categories of autosomal recessive, autosomal dominant, and X-linked. Molecularly, many of them are due to trinucleotide repeat expansions. In Friedreich ataxia, the trinucleotide repeat expansions lead to a "loss of function." In the dominant ataxias, the expanded repeats lead to a "gain of function," most likely through accumulation of intranuclear (and less commonly cytoplasmic) polyglutamine inclusions. Channelopathies can also lead to ataxia, especially episodic ataxia. Although phenotypic characteristics are an aid to the clinician, a definitive diagnosis is usually made only through genotypic or molecular studies. Genetic counseling is necessary for the testing of symptomatic and asymptomatic individuals. No effective treatment is yet available for most ataxic syndromes, except for ataxia with isolated vitamin E deficiency and the episodic ataxias. PMID- 10807079 TI - Managing the complications of cirrhosis. AB - The 3 major and potentially fatal complications of cirrhosis of the liver result from portal hypertension and include variceal bleeding, ascites, and encephalopathy. The cause of other complications, eg, thyroid dysfunction and hepatopulmonary syndrome, is uncertain. Several recent advances have occurred in the treatment of varices. However, treatment of ascites is still primarily confined to achieving a negative sodium balance, and therapy for encephalopathy centers on the use of lactulose. Although effective therapy may be available for most complications of cirrhosis, a major complication indicates a poor long-term prognosis. Liver transplantation is the only effective long-term treatment of complications due to cirrhosis. PMID- 10807080 TI - Prosthetic joint infection due to Staphylococcus lugdunensis. AB - Staphylococcus lugdunensis, a coagulase-negative staphylococcus, is being increasingly recognized as the cause of serious infections. We report 2 cases of total knee arthroplasty infection caused by S lugdunensis. S lugdunensis frequently produces a clumping factor that can result in a positive slide (short) coagulase test result. If the microbiology laboratory does not use the tube coagulase (long) test to confirm the slide coagulase test result, the organism may be misidentified as Staphylococcus aureus. S lugdunensis is more virulent than other coagulase-negative staphylococci and in many clinical situations behaves like S aureus, further increasing the confusion. However, S lugdunensis differs from S aureus in that it is susceptible to most antibiotics. This fact may alert the microbiology laboratory or the clinician that the isolate is likely not S aureus and prompt further testing of a specific isolate. Accurate identification of S lugdunensis isolates facilitates studies to define the epidemiology and pathogenesis of prosthetic joint infection due to S lugdunensis and delineates optimal medical and surgical therapies. PMID- 10807081 TI - Inguinal lymphadenitis caused by Entamoeba histolytica: case report and literature review. AB - Liver abscesses are the most common manifestation of extraintestinal infection by Entamoeba histolytica. Involvement of other sites, including the peritoneum, pericardium, brain, or genitourinary tract, is unusual. We describe a case of inguinal necrotizing lymphadenitis caused by E histolytica. Our patient responded well to surgical drainage, metronidazole, and paramomycin therapy. A literature review of genitourinary and other uncommon sites of E histolytica infection is included. PMID- 10807082 TI - Pheochromocytoma in an incidentally discovered asymptomatic cystic adrenal mass. AB - Cystic adrenal lesions can be either cortical or medullary, and distinguishing between these 2 types of lesions may be important in patient management. Pheochromocytomas, which are adrenal medullary neoplasms, typically manifest with hypertension, headaches, palpitations, tachycardia, sweating, and anxiety symptoms; however, 10% to 17% of patients with pheochromocytomas are asymptomatic. We describe a 67-year-old woman with lifelong headaches and recent persistent cough in whom a left cystic adrenal mass was incidentally discovered by computed tomography of the chest. A moderate increase in normetanephrine and total metanephrine values in two 24-hour urine samples suggested a pheochromocytoma. Computed tomography with use of contrast medium revealed ring enhancement of the cyst wall, a finding consistent with an adrenal medullary tumor. This report demonstrates the importance of repeated 24-hour urine samples to determine the metanephrine values together with contrast-enhanced computed tomography in a patient with nonspecific symptoms. PMID- 10807083 TI - Corticotropin- and thyrotropin-secreting pituitary microadenomas: detection by dynamic magnetic resonance imaging. AB - Hormone-secreting pituitary microadenomas are often not visible on magnetic resonance imaging (MRI). Diagnosis requires confirmatory endocrine test results and often an invasive procedure, inferior petrosal sinus sampling (IPSS). Improved pituitary imaging may eliminate the need for IPSS in some patients, as shown in the 2 women in this report. The first patient with hirsutism, weight gain, and hypertension had intermittent elevations of urinary free cortisol, abnormal results on a low-dose dexamethasone suppression test, and positive results on a dexamethasone-suppressed ovine corticotropin-releasing hormone test (corticotropin, increase of 122%; cortisol, increase of 118%). Gadolinium enhanced MRI showed no focal lesion, but dynamic MRI (sequential images beginning immediately after contrast injection) revealed a right-sided 5-mm microadenoma, confirmed by transsphenoidal surgery. The second patient had a goiter, anxiety, increased free thyroxine and triiodothyronine levels, and a normal thyrotropin value with no response to thyrotropin-releasing hormone. Magnetic resonance imaging showed no lesion, but dynamic MRI detected an 8-mm microadenoma. Although about 8% to 10% of healthy persons have incidental pituitary lesions that are 3 mm or larger on MRI, identification of a distinct lesion and positive results on a dexamethasone-suppressed ovine corticotropin-releasing hormone test should decrease the probability of a false-positive result on an imaging study. We recommend that dynamic MRI be performed in any patient with a suspected microadenoma, before IPSS is performed. PMID- 10807084 TI - 55-year-old woman with worsening seizure control. PMID- 10807085 TI - The mentoring partnership. PMID- 10807086 TI - Accelerated ocular hypertensive response after application of corticosteroid ointment to a child's eyelid. PMID- 10807087 TI - Role of fungi in allergic fungal sinusitis and chronic rhinosinusitis. PMID- 10807088 TI - Lactulose vs sorbitol for treatment of obstipation in hospice programs. PMID- 10807089 TI - Rehabilitative placement of poststroke patients: reliability of the Clinical Practice Guideline of the Agency for Health Care Policy and Research. AB - OBJECTIVE: To determine the interrater reliability of the United States Agency for Health Care Policy and Research (AHCPR) Clinical Practice Guideline Number 16 for rehabilitative placement of poststroke patients. DESIGN: Pairs of rehabilitation professionals, highly trained in the Guideline, rated the appropriateness of rehabilitative placements. SETTING: Acute care hospitals in three regions of the country. PATIENTS: Sixty patients with moderate-to-severe stroke. MEASURES: Numerous factors affecting appropriate placement according to the Guideline were abstracted from medical records or obtained by direct evaluation of patients. RESULTS: Good reliability was attained for home and nursing facility placement with rehabilitation services but with no multidisciplinary rehabilitation program (intraclass correlation coefficient = .73 and .60, respectively). Serious reliability problems were found for placements in low-intensity outpatient rehabilitation and high-intensity inpatient rehabilitation programs. Chief sources of unreliability were ambiguous or missing data in hospital medical records, complexities in the Guideline, and raters' tendencies to follow their own clinical judgments. More than one type of placement was appropriate for 65% of patients. CONCLUSIONS: Reliable placement guidelines are possible, but aspects of the Guideline require additional development. Evidence of demonstrated reliability and validity will be required to resolve disputes between rehabilitation professionals and payers regarding appropriate levels and types of rehabilitation and to guide patients and their families. PMID- 10807090 TI - A functional diagnostic complexity index for rehabilitation medicine: measuring the influence of many diagnoses on functional independence and resource use. AB - OBJECTIVE: To develop an index that weights diagnostic information (International Classification of Diseases, 9th revision, Clinical Modification, or ICD-9-CM codes) by the extent to which it is expected to affect functional status and inpatient rehabilitation length of stay (LOS). DESIGN: Eleven nationally prominent physiatrists assigned ICD-9-CM codes to ordinal categories by expected effect on functional recovery. A resulting functional diagnostic complexity index score was calculated for each patient by combining the ranked values of each ICD 9-CM code in the discharge record. Analyses were stratified across the 20 rehabilitation impairment categories (RICs) of the Functional Independence Measure-Function Related Groups. DATA: Patients (182,254) discharged from 465 inpatient rehabilitation units in larger hospitals and free-standing rehabilitation hospitals in 1995. RESULTS: High degrees of diagnostic complexity were associated with low levels of functional independence at admission within each of 20 RICs (Spearman R = .06 to .25). Depending on RIC, patients in the highest (compared with the lowest) complexity category had up to a 35% increase in rehabilitation LOS after accounting for the primary medical reason for rehabilitation and severity of disabilities at admission. CONCLUSION: This prototype index of medical complexity offers a promising approach for analyzing the cumulative effects of diagnoses on patients' initial functional status and rehabilitation LOS. PMID- 10807091 TI - Chest physical therapy in patients with acute exacerbation of chronic bronchitis: effectiveness of three methods. AB - OBJECTIVE: To compare the short-term effects of postural drainage (PD), oscillating positive expiratory pressure (using the FLUTTER device), and expiration with the glottis open in the lateral posture (ELTGOL) on oxygen saturation, pulmonary function, and sputum production in patients with an acute exacerbation of chronic bronchitis. DESIGN: A prospective, randomized study. SETTING: A clinical ward. PATIENTS: Ten patients with chronic bronchitis exacerbation received PD, FLUTTER, and ELTGOL by the same respiratory therapist at about the same time of day on separate days and in random order. MAIN OUTCOME MEASURES: Oxygen saturation and pulmonary function were measured before, immediately after, and 15 minutes and 1 hour after each treatment. Improvement in sputum production was measured by total sputum wet weight immediately after and for 1 hour after treatment. INTERVENTIONS: PD consisted of positioning the patients in a posture that allows bronchial drainage by gravity. FLUTTER is a device that is claimed to combine oscillating positive expiratory pressure with oscillations of the airflow. ELTGOL is an airway clearance technique that uses lateral posture and different lung volumes to control expiratory flow rate to avoid airway compression. The total time spent for treatments was 30 minutes. RESULTS: All techniques were well tolerated, and oxygen saturation and pulmonary function did not change significantly during and after treatments. Thirty minutes after the beginning of treatment, sputum production increased significantly with all techniques, but during the 1 hour after the end of treatment, it was significantly larger with FLUTTER (from 15.0 +/- 8.6g to 19.0 +/- 9.3g, p < .01) and ELTGOL (from 17.0 +/- 7.0g to 20.6 +/- 6.9g, p < .02) than with PD (from 15.5 +/- 4.0g to 17.5 +/- 3.7g, NS). CONCLUSIONS: All three treatments were safe and effective in removing secretions without causing undesirable effects on oxygen saturation, but FLUTTER and ELTGOL techniques were more effective in prolonging secretion removal in chronic bronchitis exacerbation than was the PD method. PMID- 10807092 TI - Walking symmetry and energy cost in persons with unilateral transtibial amputations: matching prosthetic and intact limb inertial properties. AB - OBJECTIVES: To investigate the hypothesis that increasing the mass and moment of inertia of the prosthetic limb of people with unilateral, transtibial amputations to match the mass and moment of inertia of the intact limb improves walking symmetry without increasing energy cost. DESIGN: Gait symmetry and metabolic energy cost of walking for six subjects with unilateral, transtibial amputations were evaluated under three prosthesis loading conditions. SETTING: University research laboratory. SUBJECTS: Six ambulatory individuals with unilateral, transtibial amputations. INTERVENTIONS: Subjects walked at 1.34 m/sec under three prosthetic limb loading conditions: (1) no added load; (2) loading that produced a match of prosthetic shank and foot mass and moment of inertia with those of the intact limb (100% load); and (3) a load that was half that of the 100% condition (50% load). MAIN OUTCOME MEASURES: Step length, swing time, stance time, and metabolic energy expenditure. RESULTS: As mass and moment of inertia of the prosthetic limb became more closely matched to the intact limb, step length, swing time, and stance time became less symmetrical. Energy cost for the 100% load condition was significantly greater (6% to 7%) than the baseline and 50% conditions. CONCLUSIONS: The loading configuration required to produce a match in the moments of inertia of the prosthetic and intact lower legs resulted in greater gait asymmetry and higher energy cost. PMID- 10807093 TI - Crutch length: effect on energy cost and activity intensity in non-weight-bearing ambulation. AB - OBJECTIVE: To investigate the effect of forearm crutch length on energy cost in three-point, non-weight-bearing (NWB) ambulation. DESIGN: Double-blind repeated measures design using crutch length as the independent variable. SETTING: Overland walking circuit at a university campus. PARTICIPANTS: Volunteer, convenience sample of 20 subjects consisting of university students and staff without cardiovascular, respiratory, or orthopedic conditions. INTERVENTION: Subjects used a three-point, NWB gait with forearm crutches set to length using conventional guidelines, and at 2.5 cm above and below this value. MAIN OUTCOME MEASURES: Oxygen consumption, carbon dioxide production, heart rate, speed of ambulation, and perceived exertion under steady-state conditions. RESULTS: In terms of oxygen cost, ambulation with crutches set to the length recommended in conventional guidelines was not significantly more energy efficient than ambulation with either the longer or shorter crutches. Using crutches set 2.5 cm longer than conventional guidelines produced the lowest respiratory exchange ratio (Vco2/Vo2) and the lowest ratings of perceived exertion. However, none of these differences reached statistical significance. CONCLUSION: Since exact crutch length was not critical in terms of oxygen cost, walking speed, or perceived exertion during NWB ambulation, the importance of rigidly adhering to specific guidelines for setting crutch length was not substantiated in this study. Clinically, consideration of patient preference regarding crutch length (within 2.5 cm) can be advocated. PMID- 10807094 TI - Reliability of measurements of pressures applied on the foot during walking by a computerized insole sensor system. AB - OBJECTIVE: To evaluate the reliability of measurements of foot pressures while walking using the F-scan computerized system with various insole sensors. DESIGN: Ten healthy subjects were studied. Each used four different pairs of insoles while walking for three 6-second epochs per insole. Peak pressures were recorded at the hindfoot, midfoot, and forefoot and analyzed. Each insole was made of pressure-sensitive 25 mm 2 cells, embedded in protective backing, and scanned electronically every 50 to 100 msec. SETTING AND PARTICIPANTS: Conventional tile floor in a gait laboratory on 10 consecutive normal subjects. RESULTS: The mean +/- standard deviation peak pressure recorded at the hindfoot was 11.1 +/- 2.0 psi; at the midfoot, 3.5 +/- 1.5 psi; and at the forefoot, 11.3 +/- 2.6 psi. The intrasubject mean differences were 0.2 psi at the hindfoot, 0.4 psi at the midfoot, and 0.3 psi at the forefoot. The data suggest that variability between different insoles is not significant. CONCLUSION: The pressure data show these insoles to be sufficiently reliable for use in patients when knowledge of foot pressure distribution is important for the relief of pain, and for designing and assessing appropriate corrective measures to relieve excessive pressure on the foot and to prevent development of pressure ulcers. PMID- 10807095 TI - Gait pattern classification of healthy elderly men based on biomechanical data. AB - OBJECTIVES: To distinguish the gait patterns of young subjects from those of elderly men using three-dimensional (3D) gait data, to determine if elderly subjects displayed other than a typical gait pattern, and to identify which parameters best describe them. DESIGN: Nonrandomized study in which video and force plate data were collected at the subject's own free walking speed and used in a 3D inverse dynamic model. Cluster analysis was chosen to identify the gait families, and analyses of variance were performed to determine which parameters were different. SETTING: A gait laboratory. PARTICIPANTS: The sample of convenience involved a single but mixed group consisting of 16 able-bodied elderly subjects (mean age, 62yrs) and 16 able-bodied young subjects aged between 20 and 35 years. MAIN OUTCOME MEASURES: Phasic and temporal gait parameters, as well as the 3D muscle powers developed in the joints of the right lower limb during the gait cycle. RESULTS: The walking patterns in elderly subjects were found to be different from those of the young adults. Three elderly gait families or groups forming a specific gait pattern were identified, and differences were found in the phasic and temporal parameters as well as in 6 peak muscle powers. Four of the peak powers occurred in the sagittal plane, and half of them were related to the hip. CONCLUSIONS: Biomechanical parameters can be used to classify the gait patterns of young and elderly men using cluster analysis rather than age alone. The muscle powers in elderly subjects are perturbed throughout the gait cycle and not only at push-off. It appears that the plane in which the peak powers occurred was related to their occurrence in the gait cycle. Variability in the gait patterns of elderly subjects could reflect natural adaptations or compensations. These should not be indicative of a deficient gait or be misconstrued as some age-related pathology. PMID- 10807096 TI - Unipedal stance testing as an indicator of fall risk among older outpatients. AB - OBJECTIVE: To test the hypothesis that a decreased unipedal stance time (UST) is associated with a history of falling among older persons. DESIGN: Fifty-three subjects underwent a standardized history and physical examination and three trials of timed unipedal stance. SETTING: The electroneuromyography laboratories of tertiary care Veterans Administration and university hospitals. SUBJECTS: Ambulatory outpatients 50 years and older referred for electrodiagnostic studies. OUTCOME MEASURES: UST and fall histories during the previous year. RESULTS: Twenty subjects (38%) reported falling. Compared with the subjects who had not fallen, those who fell had a significantly shorter UST (9.6 [SD 11.6] vs 31.3 [SD 16.3] seconds, using the longest of the three trials, p < .00001). An abnormal UST (<30sec) was associated with an increased risk of having fallen on univariate analysis and in a regression model (odds ratio 108; 95% confidence interval 3.8, >100; p < .007). The sensitivity of an abnormal UST in the regression model was 91% and the specificity 75%. When UST was considered age was not a predictor of a history of falls. CONCLUSIONS: UST of <30sec in an older ambulatory outpatient population is associated with a history of falling, while a UST of > or = 30sec is associated with a low risk of falling. PMID- 10807097 TI - Position sense testing: influence of starting position and type of displacement. AB - OBJECTIVE: To assess the effect of starting position, target position, and various types of limb displacement on repositioning tests commonly used for clinical evaluation of rehabilitation. SETTING: Controlled laboratory environment. PARTICIPANTS: Sixteen healthy volunteer subjects. MAIN OUTCOME MEASURE: Absolute error, ie, the absolute difference between target and replicate positions. DESIGN: Each subject performed four testing procedures consisting of different types of limb displacement (active, passive, and passive during antagonist muscle contraction). For each procedure, horizontal movements were performed ipsilaterally about the right glenohumeral joint from one intermediate starting position (40 degrees ) and two extreme starting positions (0 degrees and 80 degrees ). Four fixed target positions (16 degrees , 32 degrees, 48 degrees , 64 degrees ) were presented for each starting position. The subjects were required to replicate target position after returning to the respective starting position. RESULTS: Lower repositioning errors occurred with active displacement procedures compared with passive, and with the intermediate starting position compared with the extreme. Target position, however, had no effect on repositioning errors. CONCLUSIONS: Starting position and type of displacement should be considered in interpretations and comparisons of data from clinical studies. PMID- 10807098 TI - Viscosupplementation effect on proprioception in the osteoarthritic knee. AB - OBJECTIVE: To test the hypothesis that treatment of the osteoarthritic knee with intraarticular hyaluronan may improve proprioception. METHODS: Forty-six patients with unilateral osteoarthritis of the knee were recruited for this study from primary care clinics in the Department of Family Medicine at the University of Western Ontario. Inclusion required (1) the presence of pain with activities of daily function and (2) radiographic confirmation of medial compartment osteoarthritis. Proprioception was measured with an electrogoniometer attached to the study knee. Subjects performed an angle reproduction test in the closed kinetic position. The difference between the target test angle and the reproduced angle was labeled the absolute angular error (AAE). Eleven angles were tested in random order before beginning therapy, after 3 weeks, and after 12 weeks. RESULTS: Twenty-one men and 19 women completed the study (22 subjects received hyaluronan injections and 18 subjects received a placebo injection). No significant differences existed between the study groups in age, the timing of injections, or proprioceptive testing. A 2 x 3 repeated measures analysis of variance (ANOVA) comparing injection groups at all times revealed no significant differences in AAE. The AAE proprioception data was examined for differences due to fatigue caused by the number of test angles (p < .001) and differences explained by angulations in various divisions of the normal range of motion (p < .001). After accounting for these potential confounds, a two-way ANOVA still did not detect any significant differences in AAE between hyaluronan and placebo groups. CONCLUSION: Other studies have found that proprioception may be impaired in osteoarthritic knees and that viscosupplementation therapy with hyaluronan may decrease pain and increase function in these knee joints. The results of the present study suggest that this therapy does not adversely affect proprioception and that a longer, longitudinal study is required to determine if viscosupplementation treatments could attentuate proprioceptive decline. PMID- 10807099 TI - Tai Chi Chuan to improve muscular strength and endurance in elderly individuals: a pilot study. AB - OBJECTIVE: To evaluate the training effect of a Tai Chi Chuan (TCC) program on knee extensor muscular strength and endurance in elderly individuals. DESIGN: Before-after trial. SETTING: Community setting. PARTICIPANTS: Forty-one community dwelling subjects aged 61.1 +/- 9.8 years undertook a TCC program. Nine dropped out during the study. Pretraining and posttraining measurements were obtained from 15 men and 17 women. INTERVENTION: Subjects participated in a 6-month TCC program. Each session consisted of 20 minutes of warm-up, 24 minutes of structured TCC training, and 10 minutes of cool-down exercises. MAIN OUTCOME MEASURES: Peak torque of dominant and nondominant knee extensors was tested at speeds of 60 degrees , 180 degrees , and 240 degrees/sec concentrically and eccentrically. Muscular endurance of the knee extensor was tested at the speed of 180 degrees /sec. RESULTS: In the group of men, concentric knee extensor peak torque increased by 15.1% to 20.0% and eccentric peak torque increased by 15.1% to 23.7%. The group of women also showed increases, ranging from 13.5% to 21.8% in concentric peak torque, and 18.3% to 23.8% in eccentric peak torque. In addition, the knee extensor endurance ratio increased by 9.6% to 18.8% in the men and 10.1% to 14.6% in the women. CONCLUSION: TCC training may enhance muscular strength and endurance of knee extensors in elderly individuals. PMID- 10807101 TI - An extended Glasgow Coma Scale (GCS-E) with enhanced sensitivity to mild brain injury. AB - OBJECTIVE: The Glasgow Coma Scale-Extended (GCS-E) was developed to flag mild cases of concussion (corresponding to Grades I and II concussion as defined by the American Academy of Neurology) at the time of first contact with the health care system. SUBJECTS AND SETTING: The GCS-E was applied to 561 consecutive admissions with GCS scores of 13 to 15 at two hospitals in South Africa and two in the United Kingdom (UK). RESULTS: The amnesia scale was readily learned and reliably applied by emergency department staff without affecting the standard scoring of the GCS itself. Among patients with an admitting GCS of 15, 27% in the UK and 31% in South Africa reported amnesia of some duration. CONCLUSIONS: Wide use of the GCS-E would hold mild traumatic brain injury cases in the treatment loop, improve access to counselling, rehabilitation services, and personal injury compensation, and reduce the "cognitive dissonance" between victims of mild traumatic brain injury and treating professionals. PMID- 10807100 TI - Manual wheelchair pushrim biomechanics and axle position. AB - OBJECTIVE: The biomechanics of wheelchair propulsion have been linked to upper extremity injury. Specifically, prior studies have correlated increased median nerve dysfunction with increasing propulsion frequency and a higher rate of rise of the resultant, or total, pushrim force. Despite this link, there is little research on the effect of wheelchair setup on propulsion biomechanics. The objective of this study was to determine the effect of rear axle position relative to the shoulder on pushrim biomechanics. DESIGN: Case series. SETTING: Biomechanics laboratory. PARTICIPANTS: Forty individuals with paraplegia who use manual wheelchairs for mobility. INTERVENTION: Subjects propelled their own wheelchairs on a dynamometer at two different steady-state speeds and going from a dead stop to maximum speed. Bilateral biomechanical data were obtained using a force- and moment-sensing pushrim and a motion analysis system. MAIN OUTCOME MEASURES: Position of the axle relative to the shoulder at rest both horizontal (XPOS) and vertical (YPOS), and pushrim biomechanical variables including frequency of propulsion, peak and rate of rise of the resultant force, planar moment, and push angle. Partial correlation coefficients between relative axle position and propulsion biomechanics variables were calculated. RESULTS: After controlling for subject characteristics, XPOS was significantly correlated with the frequency of propulsion (p < .01) and the rate of rise of the resultant force (p < .05). In addition, both XPOS and YPOS were significantly correlated with the push angle at multiple speeds (p < .05). CONCLUSION: Specific biomechanical parameters known to correlate with median nerve injuries were found to be related to axle position relative to the shoulder. Providing wheelchair users with adjustable axle position and then fitting the user to the wheelchair can improve propulsion biomechanics and likely reduce the risk of injury. PMID- 10807102 TI - The MacNeill-Lichtenberg Decision Tree: a unique method of triaging mental health problems in older medical rehabilitation patients. AB - OBJECTIVE: Evaluation of the utility of a "decision tree" that identifies potential mental health problems in older medical patients and guides decision making for referrals. DESIGN: Measures of utility including sensitivity, specificity, and predictive power were examined. Independent t tests and nonparametric statistics were used to evaluate group differences where appropriate. SETTING: The stroke and geriatric unit of a freestanding urban medical rehabilitation hospital. SUBJECTS: In study 1,173 older, consecutively admitted medical rehabilitation patients completed all cognitive measures. In study 2, a separate sample of 313 older adults completed the Geriatric Depression Scale during admission. MAIN OUTCOME MEASURE: The MacNeill-Lichtenberg Decision Tree (MLDT) was compared with the Mini-Mental State Exam (MMSE), the Mattis Dementia Rating Scale, and the 30-item Geriatric Depression Scale. RESULTS: Study 1: The decision tree accurately triaged 87% of mental health problems and allowed for deferral of 41% of cases, for whom further assessment was unnecessary. The MLDT was superior to the MMSE, with higher sensitivity and a lower failure rate. Study 2: The emotional status component of the MLDT was useful in triaging cases for depression evaluation. CONCLUSION: The MLDT was useful in prioritizing cases with regard to mental health problems (eg, dementia, depression) and making quick referral decisions. The MLDT is a unique instrument that not only evaluates cognitive status, but also considers psychosocial factors and emotional status in older adults. PMID- 10807103 TI - Functional recovery in cancer rehabilitation. AB - OBJECTIVE: To assess the impact of inpatient rehabilitation on the motor and cognitive functional status of cancer patients, and to determine whether cancer diagnosis, rehabilitation impairment, physician-determined rehabilitation goals, and active cytotoxic treatment affect the magnitude of functional improvement. DESIGN AND SETTING: A retrospective, case series of patients with an oncology diagnosis undergoing inpatient rehabilitation at a rehabilitation hospital. PARTICIPANTS: A sample of 200 patients admitted for rehabilitation services due to disability resulting from impairments caused by cancer or its treatment. INTERVENTION: Comprehensive multidisciplinary inpatient rehabilitation. OUTCOME MEASURES: Function status was measured using the motor and cognitive measures of the Functional Independence Measure. RESULTS: All patients made significant gains in motor function regardless of diagnostic group, rehabilitation impairment group, rehabilitation goal group, and cytotoxic treatment status. The magnitude of motor function gain was not equivalent across all impairments and rehabilitation goals. Significant gains in cognitive function were made by all patients except those with intracranial neoplasms, central nervous system dysfunction, and palliative rehabilitation goals. CONCLUSION: Inpatient rehabilitation can improve both motor and cognitive function in patients with disability resulting from impairments caused by cancer or its treatment. PMID- 10807104 TI - Assessment training and team functioning for treating children with disabilities. AB - OBJECTIVE: To investigate differences in team functioning before and after assessment training. DESIGN: Before-after trial. SETTING: Outpatient rehabilitation and education program in a university medical center-based rehabilitation institute. PARTICIPANTS: A population-based sample of 19 rehabilitation and education specialists. INTERVENTION: Baseline measures were taken during four meetings using the multidisciplinary team approach. Training in a global measure of child development ensued and staff administered the test to children in their care. Test results were then used during four transdisciplinary team meetings. OUTCOME MEASURES: Behavioral ratings of team participation (Transdisciplinary Team Rating Scale), self-report instruments of team development (Team Assessment Questionnaire), treatment planning and goal development (Staff Perception Questionnaire). RESULTS: Results of t tests confirmed two of three hypotheses. There was more team member participation during transdisciplinary meetings than during multidisciplinary meetings (p = .003) and staff members' perceptions of the efficacy of treatment planning and implementation of goals significantly favored the transdisciplinary model (p < .001). No significant difference was found in team development (p = .254). CONCLUSION: This study provides evidence for the value of training in an assessment tool that requires the use of a common, functional language that is comprehensible to all staff members and promotes a transdisciplinary approach to team work. PMID- 10807105 TI - A therapy-relevant casemix classification system for school-age children with disabilities. AB - OBJECTIVE: To develop a clinically meaningful classification system of resource homogeneous groups to describe therapy resource use for school-age children with disabilities. DESIGN: Work-time allocation survey of therapy services (physical therapists, occupational therapists, and speech pathologists). SETTING: Three main disability agencies in Western Australia. CLIENTS: All children of two agencies, a random sample of children of the third. A total of 644 clients and their services were studied. MAIN OUTCOME MEASUREMENT: Minutes of therapy service time expected over a 10-week school term. RESULTS: Forty-six percent of the variance in a measure of time spent with clients can be explained using a classification system of nine groups, based on splitting a mobility measure (4 major groups), then the nature of primary disorder (2 subgroups), independence in self care (3 subgroups), and expressive communication (3 subgroups). CONCLUSION: This study defines a preliminary classification system for the distribution of therapy resources to school-age children with disabilities. This model has the potential to be used to purchase services on a fairer basis than traditional, historical funding methods have allowed. PMID- 10807106 TI - Classifying incomplete spinal cord injury syndromes: algorithms based on the International Standards for Neurological and Functional Classification of Spinal Cord Injury Patients. AB - OBJECTIVE: To develop an objective and uniform means for classifying patients with incomplete spinal cord injury (SCI) according to SCI syndromes. DESIGN: Criteria for assigning the syndromes (defined by the International Standards for Neurological and Functional Classification of SCI Patients) were operationalized by means of sensory and motor scores and were incorporated into a set of six independent algorithms and two composite algorithms. SETTING: A regional SCI rehabilitation center in Canada. PATIENTS: SCI patients (n = 56) with incomplete injuries (American Spinal Injury Association classes B, C, D) and stable neurologic deficits. RESULTS: Individual algorithms allowed the highest classification rate but with some patients meeting the criteria for more than one syndrome. A composite, differential allocation algorithm, with selected thresholds at decision nodes, yielded a classification rate approximating that of the individual algorithms but without double classifications. CONCLUSIONS: The composite algorithm provided an objective and standardized means of assigning patients to syndromes based on clinically measurable sensory and motor scores. The thresholds used to implement criteria and the order of decision nodes greatly influenced the outcomes and may be adjusted to suit the needs of the classification, that is, embracing liberal or stringent criteria. Controversy remains about the interpretation of some syndromes, and many patients remain unclassifiable because of mixed clinical presentation. PMID- 10807107 TI - Muscle force measured using "break" testing with a hand-held myometer in normal subjects aged 20 to 69 years. AB - OBJECTIVE: To measure the strength of 17 muscle groups in the upper and lower extremities in a large group of healthy subjects using "break" testing with a hand-held myometer, and to examine the intrasession and intersession reliability of the testing protocol. SUBJECTS AND INSTRUMENTATION: A convenience sample of 20 men and 20 women in each decade of age from 20 to 69 years (n = 200) was tested using a Penny & Giles hand-held myometer. RESULTS: Reliability coefficients were >.85 for both intrasession and intersession reliability, except for the ankle dorsiflexors. Men exerted a significantly greater force than women for all muscle groups. Age, weight, and side of testing were significant predictors of force in the majority of muscle groups. The fifth percentile values, as the lower limit of normal, are reported separately for gender and side of testing for each decade of age. CONCLUSION: Using the testing protocol specified in this study, data from patients with various neuromuscular diseases may be compared with the appropriate gender- and age-matched normal data to accurately identify the presence of weakness. PMID- 10807108 TI - Complementary and alternative medicine: Canadian physiatrists' attitudes and behavior. AB - OBJECTIVE: To document the prevalence and patterns of knowledge about, referrals to, training in, and practice of complementary and alternative medical therapies and their perceived effectiveness by a sample of Canadian physiatrists. DESIGN: Cross-sectional survey by written questionnaire. SETTING: Physiatrists in the province of Ontario, Canada. SUBJECTS: All 116 physiatrist/rehabilitation specialists listed in the Ontario Medical Association directory. Data were obtained from 98 respondents. MAIN OUTCOME MEASURES: Statistical analysis of responses in these areas: attitudes, knowledge, and recommendations about alternative therapies, and clinical approach including referral pattern, training, and practice of alternative medicine. RESULTS: Of the respondents, 72% reported referring patients for alternative medicine therapies, and 20% had training in and 20% practiced some form of alternative medicine. The therapies rated highest in usefulness were acupuncture (85%), biofeedback (81%), and chiropractic (80%). Sixty-three percent believed that alternative medicine had ideas and methods that would be of benefit to physiatrists. Only 9% believed it to be a threat to public health. A greater proportion of physiatrists who refer were women, were younger, had graduated more recently, and scored higher in their ratings of more useful alternative medicine therapies. Previous training in alternative medicine was correlated with a higher practice rate but not with referral rate. Practice profile and academic affiliation were not associated with greater or less use of alternative medicine. CONCLUSION: In Ontario, physiatrists report greater knowledge of and more use of alternative medicine therapies than do general practitioners. The most commonly used therapies are acupuncture, biofeedback, and manipulation (chiropractic, osteopathy). It is recommended that these approaches be taught in physiatry residency training and be the focus of future research. Incorporating such therapies into practice will help to meet the public demand for such approaches in the decade to come. PMID- 10807109 TI - Ginkgo biloba extract: mechanisms and clinical indications. AB - OBJECTIVE: Ginkgo biloba may have a role in treating impairments in memory, cognitive speed, activities of daily living (ADL), edema, inflammation, and free radical toxicity associated with traumatic brain injury (TBI), Alzheimer's dementia, stroke, vasoocclusive disorders, and aging. The purpose of this review is to provide a synthesis of the mechanisms of action, clinical indications, and safety of Ginkgo biloba extract. DATA SOURCES: Empirical studies, reviews, chapters, and conference proceedings were identified in the following databases: Medline, the Research Council for Complementary Medicine based on the British Library database, and Psychlnfo. Ginkgo biloba, EGb 761, Tanakan, Tebonin, Rokan, and LI 1370 were the principal index terms. STUDY SELECTION AND DATA EXTRACTION: Controlled clinical studies with both positive and negative findings are included, in addition to animals studies illustrating mechanisms of activity. DATA SYNTHESIS: Ginkgo has shown activity centrally and peripherally, affecting electrochemical, physiologic, neurologic, and vascular systems in animals and humans with few adverse side effects or drug interactions. Ginkgo shows promise in patients with dementia, normal aging, and cerebrovascular-related disorders. Clinical indications include memory, information processing, and ADL. CONCLUSIONS: Ginkgo shows promise in treating some of the neurologic sequelae associated with Alzheimer's disease, TBI, stroke, normal aging, edema, tinnitus, and macular degeneration. Mechanisms of action may include antioxidant, neurotransmitter/receptor modulatory, and antiplatelet activating factor properties. While safe, caution is advised when recommending ginkgo to patients taking anticoagulants. Future studies should examine dose effects, component activity, mechanisms, and clinical applications. PMID- 10807110 TI - Shoulder impingement in tennis/racquetball players treated with subscapularis myofascial treatments. AB - Conservative care of the athlete with shoulder impingement includes activity modification, application of ice, nonsteroidal anti-inflammatory drugs, subacromial corticosteroid injections, and physiotherapy. This case report describes the clinical treatment and outcome of three patients with shoulder impingement syndrome who did not respond to traditional treatment. Two of the three were previously referred for arthroscopic surgery. All three were treated with subscapularis trigger point dry needling and therapeutic stretching. They responded to treatment and had returned to painless function at follow-up 2 years later. PMID- 10807111 TI - Antegrade continence enema for the treatment of neurogenic constipation and fecal incontinence after spinal cord injury. AB - OBJECTIVE: To describe the effects of an antegrade continence enema stoma formed in a paraplegic man with intractable constipation and fecal incontinence. DESIGN: Case report. SETTING: Spinal cord injury unit, Veterans Affairs hospital. PARTICIPANTS: Spinal cord injury (SCI) patient with T12 paraplegia. INTERVENTION: Surgical formation of antegrade continence enema stoma. MAIN OUTCOME MEASURES: Time of bowel program care, ease of fecal elimination, safety of procedure. RESULTS: Bowel care time was decreased from 2 hours to 50 minutes daily; 6 bowel medications were discontinued; fecal incontinence was eliminated; and no surgical or medical side effects noted after the procedure. CONCLUSION: The antegrade continence enema procedure is a safe and effective means of treating intractable constipation and fecal incontinence in the adult SCI patient. This option should be considered for those persons in whom medical management of bowel care has been unsuccessful. PMID- 10807112 TI - Relation of impairment to everyday competence in visual disorientation syndrome: evidence from a single case study. AB - OBJECTIVE: To determine the relation of neurology and neuropsychology to everyday competence. DESIGN: The association of these three domains was investigated using a single case multiple baseline design with two phases. Phase A comprised 6 weeks that coincided with an inpatient admission. Phase B comprised 3 months spent at home. A battery of visual spatial tests was completed every fortnight during the A phase and at the end of the B phase. Two new tests of relevant neurologic function with control data were developed and used weekly during the A phase and at the end of the B phase. The first test recorded the speed, accuracy, and efficiency of her walking, and the second test recorded her depth perception. SETTING: Tertiary care center. PARTICIPANT: A 35-year-old woman who suffered a venous sinus thrombosis with visual disorientation syndrome. RESULTS: During Phase A, she achieved significant functional gains in mobility, dressing, bathing, and domestic tasks, in the context of unchanging psychometric test scores and static relevant neurologic function. During Phase B, she achieved few functional gains, despite improvements in neurologic status, demonstrated by depth perception. CONCLUSIONS: Everyday function can progress without improvement in neurologic and cognitive status. PMID- 10807113 TI - Treatment with subantimicrobial dose doxycycline improves the efficacy of scaling and root planing in patients with adult periodontitis. AB - BACKGROUND: In a previous study, subantimicrobial dose doxycycline (SDD) significantly improved clinical parameters associated with periodontal health in patients with adult periodontitis (AP) when used as an adjunct to a maintenance schedule of supragingival scaling and dental prophylaxis. In this double-blind, placebo-controlled, parallel-group, multicenter study, the efficacy and safety of SDD were evaluated in conjunction with scaling and root planing (SRP) in patients with AP. METHODS: Patients (n = 190) received SRP at the baseline visit and were randomized to receive either SDD 20 mg bid or placebo bid for 9 months. Efficacy parameters included the per-patient mean changes in clinical attachment level (CAL) and probing depth (PD) from baseline, the per-patient percentages of tooth sites with attachment loss (AL) > or = 2 mm and > or = 3 mm from baseline, and the per-patient percentage of tooth sites with bleeding on probing. Prior to analysis, tooth sites were stratified by the degree of disease severity evident at baseline RESULTS: In tooth sites with mild to moderate disease and severe disease (n = 183, intent-to-treat population), improvements in CAL and PD were significantly greater with adjunctive SDD than with adjunctive placebo at 3, 6, and 9 months (all P <0.05). In tooth sites with severe disease, the per-patient percentage of sites with AL > or = 2 mm from baseline to month 9 was significantly lower with adjunctive SDD than with adjunctive placebo (P<0.05). Improvements in clinical outcomes occurred without detrimental shifts in the normal periodontal flora or the acquisition of doxycycline resistance or multiantibiotic resistance. SDD was well tolerated, with a low incidence of discontinuations due to adverse events. CONCLUSIONS: The adjunctive use of SDD with SRP is more effective than SRP alone and may represent a new approach in the long-term management of AP. PMID- 10807114 TI - Cyclic tension force activates nitric oxide production in cultured human periodontal ligament cells. AB - BACKGROUND: Nitric oxide (NO) is involved in a number of physiological and pathophysiological processes. The aim of this study was to examine the ability of human periodontal ligament (PDL) cells to produce NO and whether mechanical forces could induce NO production in the PDL cells. METHODS: Human PDL cells were seeded onto flexible bottoms of a culture plate and subjected to cyclic tension forces. NO production was evaluated by measuring concentration of NO2- and NO3- (NO2-/NO3-), the oxidized products of NO, in the culture medium. We employed reverse transcription polymerase chain reaction (RT-PCR) methods to detect NO synthase mRNA in the PDL cells. NO synthase immunoreactivity was also evaluated in both stimulated and unstimulated PDL cells. RESULTS: In unstimulated PDL cell culture, NO2-/NO3- increased to 140% of the initial value in 12 hours. In contrast, NO2-/NO3- showed a 3-fold increase when the cells had been subjected to cyclic tension forces for 12 hours. The increase in NO production was blocked by NG-monomethyl-L-arginine (5 x 10(-4) M), an inhibitor of NO synthase. Endothelial NO synthase (ecNOS) mRNA was expressed in both stimulated and unstimulated PDL cells, whereas inducible NO synthase (iNOS) mRNA was detected in neither culturing condition. We found strong ecNOS but not iNOS immunoreactivity in the stimulated PDL cells. CONCLUSIONS: These results suggest that human PDL cells produce NO by ecNOS and that the production is enhanced by stimulating the cells with cyclic tension forces. Mechanically stimulated PDL cells may modulate the function of periodontium by the upregulated NO production. PMID- 10807115 TI - Effect of a ceramic and a non-ceramic hydroxyapatite on cell growth and procollagen synthesis of cultured human gingival fibroblasts. AB - BACKGROUND: Ceramic hydroxyapatites and non-ceramic hydroxyapatites have been used extensively as alloplastic materials for bone reconstruction. However, different forms of hydroxyapatite induce different types of tissue response. METHODS: Human gingival fibroblasts (FMM1 cells) were used to analyze ceramic and non-ceramic hydroxyapatite biocompatibility. The cells were grown on surfaces covered either by collagen (control group), collagen plus ceramic hydroxyapatite, or collagen plus non-ceramic hydroxyapatite. Scanning electron microscopy, growth and cell viability curves, and procollagen immunoprecipitation were obtained. For the growth and viability curves, 10(4) cells were seeded on 60 mm dishes. Cells from each group were counted, in triplicate, at 1, 3, 4, 5, and 6 days after seeding using the Trypan blue dye exclusion assay. RESULTS: The cells grew in close contact with both types of hydroxyapatite particles. No differences were found in the amount of procollagen synthesis among any experimental group. The cultures treated with ceramic hydroxyapatite had a growth delay for the first 5 days. There was no difference in cell viability between the control group and the non-ceramic hydroxyapatite group. However, cultures treated with ceramic hydroxyapatite showed significantly lower viability percentages than the other groups. CONCLUSIONS: Hydroxyapatite supports cell growth and fibroblast metabolism including collagen production, and hence is biocompatible. Cell viability and structural studies showed that non-ceramic hydroxyapatite has relevant physical and biological properties as an implant material. PMID- 10807116 TI - The effect of inter-implant distance on the height of inter-implant bone crest. AB - BACKGROUND: The biologic width around implants has been well documented in the literature. Once an implant is uncovered, vertical bone loss of 1.5 to 2 mm is evidenced apical to the newly established implant-abutment interface. The purpose of this study was to evaluate the lateral dimension of the bone loss at the implant-abutment interface and to determine if this lateral dimension has an effect on the height of the crest of bone between adjacent implants separated by different distances. METHODS: Radiographic measurements were taken in 36 patients who had 2 adjacent implants present. Lateral bone loss was measured from the crest of bone to the implant surface. In addition, the crestal bone loss was also measured from a line drawn between the tops of the adjacent implants. The data were divided into 2 groups, based on the inter-implant distance at the implant shoulder. RESULTS: The results demonstrated that the lateral bone loss was 1.34 mm from the mesial implant shoulder and 1.40 mm from the distal implant shoulder between the adjacent implants. In addition, the crestal bone loss for implants with a greater than 3 mm distance between them was 0.45 mm, while the implants that had a distance of 3 mm or less between them had a crestal bone loss of 1.04 mm. CONCLUSIONS: This study demonstrates that there is a lateral component to the bone loss around implants in addition to the more commonly discussed vertical component. The clinical significance of this phenomenon is that the increased crestal bone loss would result in an increase in the distance between the base of the contact point of the adjacent crowns and the crest of bone. This could determine whether the papilla was present or absent between 2 implants as has previously been reported between 2 teeth. Selective utilization of implants with a smaller diameter at the implant-abutment interface may be beneficial when multiple implants are to be placed in the esthetic zone so that a minimum of 3 mm of bone can be retained between them at the implant-abutment level. PMID- 10807117 TI - Pocket oxygen tension in smokers and non-smokers with periodontal disease. AB - BACKGROUND: Smoking is associated with destructive periodontal disease. Pocket oxygen tension (pO2) is likely to be a major environmental determinant of the subgingival microflora, which is a primary etiological factor of the disease. This study aimed to compare the pocket pO2 in smokers and non-smokers with periodontal disease. METHODS: Pocket oxygen tension was compared in 27 smokers and 34 non-smokers by considering 2 confounding factors, probing depth and oxygen saturation of hemoglobin (S(O2)), in the gingiva. The pO2 was determined using oxygen microelectrode by polarographic method with an electronic compensation circuit for subgingival temperature. Gingival S(O2) was determined using tissue reflectance spectrophotometry. RESULTS: No significant difference was found in the modified gingival index and the plaque index between smokers and non-smokers. The pO2 was significantly lower in smokers (21.9+/-9.6 mmHg) than in non-smokers (33.4+/-8.4 mmHg). The difference was highly significant (P <0.0001) and was consistent when the confounding factors were considered. Correlation between the PO2 and probing depth approached statistical significance in smokers (r = -0.36, P = 0.0674) and significance in non-smokers (r = -0.41, P = 0.0174). Correlation of the PO2 to the gingival S(O2) was highly significant in non-smokers (r = 0.57, P = 0.0005), but no association was found in smokers (r = -0.08, P= 0.6975). CONCLUSIONS: These findings indicate that pO2 is lower in smokers than in non smokers, and that the pO2 in smokers is not influenced by gingival oxygen sufficiency. The present study may provide the basis of understanding environmental factors possibly associated with microbial flora in the pockets of smokers. PMID- 10807118 TI - Outcome variables in periodontal research: means and threshold-based site changes. AB - BACKGROUND: The exclusive use of mean measurements in periodontal research might at times be misleading, as changes in different sites in the same individual might nullify each other. The purpose of the present study was to compare disease progression and response to periodontal therapy using both individual site activity with thresholds and mean patient changes. METHODS: Seventy-nine (79) subjects with established periodontitis were monitored for 1 year (no treatment [NTx] group); 108 subjects who received scaling, root planing, and quarterly prophylaxis were observed in a similar time interval (treatment [Tx] group). Probing depth (PD), attachment level (AL) and alveolar crestal height (ACH) were measured at baseline and 1 year using pressure-sensitive probes and computer assisted image analysis of radiographs. RESULTS: Mean reduction in PD (0.50 mm) was observed in the Tx group compared to a small increase (-0.04 mm) in the NTx group (P= 0.0001). Treatment resulted in mean AL gain (0.44 mm) compared to net AL loss (-0.21 mm) in the NTx group (P= 0.0001). Subjects in the Tx group had twice as many sites with AL gain (beyond the threshold) compared to NTx subjects (16.64+/-1.07% versus 8.11+/-0.68%) which was highly significant (P = 0.0001, Student t test). Similarly, although in the opposite direction, percentage of sites with AL loss beyond threshold was 6.21+/-0.47% (Tx group) and 14.02+/-1.15% (NTx group) which was also highly significant (P = 0.0001, Student t test). Change in ACH was minimal for the Tx group (-0.07+/-0.03 mm) while NTx subjects experienced greater bone loss throughout the year (-0.16+/-0.02 mm); percentage of sites with ACH loss was similar for both groups, while the Tx group had 3 times the number of sites with ACH gain (11.76% versus 3.42%), suggesting that the reduction in mean bone loss in the Tx group did not result from a reduction in losing sites, but rather from an increase in gaining sites. The use of both means and binary data seems to supplement the information regarding the nature and mechanism of disease progression and arrest. CONCLUSIONS: The characteristics of periodontal disease with its multifactorial patient, local, and site-related etiology support the use of both mean and site-based changes in clinical studies of periodontal disease and treatment. PMID- 10807119 TI - The prevalence of pathogenic periodontal microflora in healthy young adult smokers. AB - BACKGROUND: Smoking is a major risk factor in periodontitis, although the mechanisms of its effects are not well understood. The overall goal of this clinical study was to determine if smoking enhances the colonization of the oral cavity by pathogenic bacteria in a periodontitis-free population. The prevalence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Eikenella corrodens, Bacteroides forsythus, and Treponema denticola was investigated in 25 smokers and 25 non-smokers by using DNA probes. METHODS: The subjects were 21 to 35 years of age with a healthy periodontium or slight gingivitis and were systemically healthy. The test group included subjects who had a minimum of a 1.5 pack-year history of smoking, while the control subjects never smoked. Subgingival plaque samples were taken by paper point following the assessment of multiple clinical parameters. RESULTS: This investigation showed: 1) no statistically significant differences were noted in any clinical parameter measured between the groups; 2) of the 8 subjects who were infected by at least 1 tested pathogen, seven were smokers (P= 0.02); 3) infected smokers had a 15.7+/-3.5 pack-year history and smoked a mean of 27+/-5 cigarettes/day versus 4.4+/-0.8 pack years and 15+/-1 cigarettes/day for the non-infected smokers (P = 0.0001 and P = 0.004); and 4) smokers were 18 times more likely to exhibit the presence of pathogens than non smokers. CONCLUSIONS: These data indicate that the prevalence of colonization of the sulcus by pathogenic bacterial species in periodontitis-free individuals is related to the quantity and duration of cigarette smoking. PMID- 10807120 TI - Citric acid demineralization and subepithelial connective tissue grafts. AB - BACKGROUND: The present study was designed to evaluate the effect, if any, of citric acid root demineralization in the outcome of subepithelial connective tissue grafts performed to cover localized gingival recessions. METHODS: Thirty six patients participated, each providing one gingival recession; 19 received citric acid demineralization, while 17 did not. All were treated surgically with subepithelial connective tissue grafts and followed for 6 months. At baseline and 6 months, the following parameters were recorded: plaque index, gingival index, recession height, probing depth, recession width, and amount of keratinized tissue. Data were analyzed statistically to 1) evaluate the results achieved with each procedure individually over time and 2) compare the results obtained with the 2 procedures after 6 months. RESULTS: Results showed significant reductions in recession height after 6 months independently of whether citric acid was applied or not (2.79+/-0.79 versus 2.56+/-0.73). Similarly, recession width was significantly reduced (3.74+/-1.19 versus 3.50 +/-0.73), and the width of keratinized tissue was significantly increased (2.47+/-1.6 versus 2.3+/-1.2). No significant changes in probing depth were found (-0.16+/-0.06 versus -0.13+/ 0.81). No significant differences were found when both techniques were compared in any one of the parameters analyzed (all P >0.30). CONCLUSIONS: It is concluded that: 1) the subepithelial connective tissue graft procedure provides a satisfactory solution in the treatment of localized gingival recessions, and 2) citric acid demineralization does not affect the clinical outcome of the surgical technique. PMID- 10807121 TI - The effect of interleukin-11 on the progression of ligature-induced periodontal disease in the beagle dog. AB - BACKGROUND: The rate of progression of periodontal disease is dependent on the complex regulatory interactions between bacteria and the immune modulators of the host response. The purpose of this investigation was to determine if recombinant human interleukin-11 (rhIL-11), known to downregulate several inflammatory modulators, has the ability in subcutaneous administration to reduce the rate and/or extent of periodontal attachment loss and radiographic bone loss in a ligature-induced periodontal disease beagle dog model. METHODS: Twenty 18-month old female beagle dogs were brought to optimal periodontal health over a 2-week period. Periodontal disease was induced by placing 2.0 silk ligatures around the mandibular first molar and premolar teeth. The dogs were divided into 3 treatment groups and one control group. The 3 treatment groups received subcutaneous injections of either 15, 30, or 80 microg/kg of rhIL-11 in saline buffer twice a week. The placebo group received buffer only subcutaneously twice a week. The gingival health of each animal was measured by recording the presence or absence of gingival inflammation, plaque, and bleeding upon probing. Attachment levels and bone height were also measured. Treatment administration and clinical and radiographic evaluations were performed in a masked fashion. RESULTS: At week 8, the placebo group had 3.89 mm of attachment loss and 73.8% radiographic bone remaining. The 15 microg/kg group had 1.99 mm attachment loss and 89.5% bone remaining; the 30 microg/kg group had 0.84 mm attachment loss and 92.5% bone remaining; and the 80 microg/kg group had 1.05 mm attachment loss and 85.5% bone remaining. All 3 treatment groups lost significantly less attachment and retained significantly more bone than did the placebo group. CONCLUSIONS: The study indicates that subcutaneous injections of rhIL-11 were able to slow the progression of attachment and radiographic alveolar bone loss in a ligature induced beagle dog model. PMID- 10807122 TI - Periodontal treatment decisions for molars: an analysis of influencing factors and long-term outcome. AB - BACKGROUND: The purpose of this retrospective study was 1) to analyze factors influencing the treatment decision for periodontitis-affected molars and 2) to evaluate the long-term outcome of the decision. The treatment alternatives included in the analysis were maintenance of the tooth with only non surgically/surgically performed scaling/root planing with or without furcation plasty, root separation/resection, and tooth extraction. METHODS: Data collected from a total of 1,313 first and second molars in 222 patients (mean age 45 years; range 14 to 73 years), referred for periodontal treatment, were analyzed with respect to treatment decisions for furcation-involved teeth. The patients had been examined with respect to oral hygiene status, gingival conditions, probing depth, furcation involvement, and radiographic bone height before and after active periodontal treatment. Frequencies, mean values and standard deviations were calculated for the various variables assessed. Multiple regression models were formulated in order to analyze the influence of various variables on treatment decisions. One-hundred and sixty (72%) of the patients agreed to participate in a follow-up examination to determine the clinical and radiographic status of the molars 8 to 12 years after the active phase of treatment for evaluation of the long-term outcome of the treatment decision. RESULTS: Twenty eight percent of the molars were extracted and 4% were root separated/resected. Factors found to significantly influence the decision of tooth extraction were tooth mobility, tooth position, lack of occlusal antagonism, degree of furcation involvement, and remaining bone support. Of the factors analyzed with respect to root separation/resection, molar position and degree of furcation involvement showed the strongest influence on the treatment decision. The 8- to 12-year follow-up examination revealed that 96% of the molars subjected to non-resective therapy were still in function. The corresponding figure for root separated/resected molars was 89%. Loss of radiographic bone support during the 10-year follow-up period was 0.1 to 0.6 mm for the various molars with the highest value for maxillary second molars. CONCLUSIONS: A conservative approach to the treatment of molars with even deep furcation invasions may show a high long-term success rate, provided maintenance care is offered. PMID- 10807123 TI - Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts. AB - BACKGROUND: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. METHODS: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiaton (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-beta1 and prostaglandin E2 (PGE2) compared. RESULTS: Profilometry showed the polished and TCN surfaces were smooth with comparable Ra values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC-treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-beta1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. CONCLUSIONS: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-beta1. However, later differentiation events like osteocalcin production are decreased. Osteoclast-mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces. PMID- 10807124 TI - Clinical validation of a new subtraction radiography technique for periodontal bone loss detection. AB - BACKGROUND: Diagnostic subtraction radiography (DSR) is a new digital radiographic image subtraction method designed to enhance detection of crestal or periapical bone density changes and to help evaluate caries progression in teeth. In this clinical study, the performance of the DSR method was evaluated for its ability to detect periodontal bone loss and was compared with that of conventional evaluation of radiographs and the standardized cephalostat-guided image acquisition and subtraction technique (LRA) which served as the "gold standard." METHODS: In each of 25 subjects with alveolar crestal bone loss created by periodontal surgery, one set of DSR radiographs and one set of LRA radiographs were obtained before and after the surgery. Subtraction images were then generated by both the proprietary DSR and the LRA techniques. Four viewers evaluated the paired film sets and both subtraction image sets using a 5 point confidence scale to determine the presence or absence of crestal bone loss. Receiver operating characteristics (ROC) statistical procedures were applied to analyze the diagnostic accuracy and statistical differences between the three imaging modalities. RESULTS: The DSR subtraction viewing generated an ROC area of 0.882. For 2 of the viewers this represented a statistically significant gain (P <0.05) over the conventional viewing of the radiographs which had an average ROC area of 0.730. In comparison, the LRA method achieved an area of 0.954. The differences between the LRA and the DSR subtraction methods were not statistically significant, but the statistical power for claiming equality was low ranging from 0.2 to 0.6. CONCLUSIONS: The use of the DSR technique in clinical radiographic image acquisition and subsequent subtraction analysis clearly enhanced the accuracy of alveolar crestal bone loss detection when compared to conventional film viewing. Because this methodology is less resource demanding than LRA and the film exposure techniques and computer-based image analysis skills may be acquired with only a few hours of training, the DSR has potential in clinical practice. PMID- 10807125 TI - Interleukin-1 gene polymorphisms and long-term stability following guided tissue regeneration therapy. AB - BACKGROUND: Specific interleukin (IL)-1 gene polymorphisms are associated with an increased susceptibility to severe periodontitis, increased inflammation, and increased likelihood of tooth loss during the maintenance phase after conventional periodontal therapy. The aim of the present study was to evaluate the impact of genotype on the maintenance of gained clinical attachment obtained after guided tissue regeneration (GTR) surgical therapy in deep intrabony defects. METHODS: Forty deep (> or =4 mm) interproximal angular bony defects with presurgical clinical attachment loss of >8 mm were treated by GTR using a non absorbable expanded polytetrafluoroethylene (ePTFE) membrane. Membranes were surgically removed 4 to 6 weeks after surgery. Afterwards patients were placed on monthly recall for the first year and every 3 months for the following 3 years. At the 4-year re-evaluation, a IL-1 genetic susceptibility test was performed on all patients. RESULTS: Fourteen (35% of the 40 patients) were genotype-positive (+). At baseline no statistically significant differences were found between patients with different genotypes in full mouth plaque score (FMPS), full mouth bleeding score (FMBS), clinical attachment level (CAL), probing depth (PD), or gingival recession. At year 1 follow up visit, no statistically significant differences were noted between genotype + and genotype - patients in FMPS, FMBS, amount of CAL gain, decrease in PD, or increase in gingival recession. Sixteen patients had membrane exposure after the GTR procedures. In these patients, the amount of CAL gain (P <0.001) and PD reduction (P <0.01) 1 year after surgery was significantly lower than those observed in patients without membrane exposure. At the year 4 follow-up visit, no significant differences were found between genotype negative and positive patients in FMPS or FMBS and both groups showed a significant loss in CAL (P<0.001) and increase in PD (P<0.001) when compared to year 1 visit. No change in gingival recession was noted. Genotype + patients showed significantly more CAL loss (P<0.002) and increase in PD (P<0.001) between the years 1 and 4 when compared to genotype - patients. A significant association between genotype and stability of the regenerated attachment was also demonstrated. CONCLUSIONS: The results of this study demonstrate that genotype expression did not effect GTR treatment response at 1 year, but had a great impact on long-term stability (year 4). In a 3-year period, patients with positive IL-1 genotype lost about 50% of the first year gained CAL and were about 10 times more likely of experiencing > or = 2 mm CAL loss when compared to oral hygiene matched genotype-negative patients. PMID- 10807126 TI - Expression of molecules involved in antigen presentation and T cell activation (HLA-DR, CD80, CD86, CD44 and CD54) by cultured human osteoblasts. AB - BACKGROUND: Osteoblasts express the CD44 antigen and HLA class II antigens, molecules which, together with other costimulatory molecules such as CD80, CD86, and CD54, are involved in antigen presentation and T cell activation. The aim of this study was to investigate the expression of these molecules in human osteoblasts. METHODS: Human osteoblastic cells obtained from samples of normal bone obtained during mandibular osteotomy were isolated, maintained in culture, and characterized. The identity of the cells was confirmed by their alkaline phosphatase activity and their capacity to produce osteocalcin. Flow cytometry was used to examine the expression HLA-DR, CD80, CD86, CD44, and CD54 molecules involved in immune activities. RESULTS: We detected the expression of CD10, CD44, and HLA-DR antigens, molecules involved in antigen presentation in cultured osteoblastic cells. Although the cells were negative for CD45, the leukocyte common antigen and CD14 (an antigen detected on macrophages), they expressed CD54, CD80, and CD86 antigens, which are also involved in the mechanisms of antigen presentation to and activation of T cells. CONCLUSIONS: Our results suggest that osteoblastic cells or a subpopulation of these cells may have immune functions in bone. Further studies in which immune functions are assessed will be needed to test this hypothesis. PMID- 10807127 TI - Analysis of human gingival tissue and gingival crevicular fluid beta glucuronidase activity in specific periodontal diseases. AB - BACKGROUND: Beta-glucuronidase (betaG) is one of the enzymes involved in the destruction of non-collagenous components of the extracellular matrix. It is also considered an indicator or predictor of periodontal disease activity. The present study was conducted to determine the presence and the levels of betaG activity in gingival tissue and gingival crevicular fluid (GCF) in periodontal disease and health status. The validity of 2 expressions of data, total betaG activity versus betaG concentration, and the correlations between clinical periodontal status and betaG profile was also evaluated. METHODS: betaG activities in gingival tissues and GCF samples from 57 individuals, divided into 3 equal groups of adult periodontitis (AP), early-onset periodontitis (EOP), and periodontally healthy subjects were spectrophotometrically examined. RESULTS: Both patient groups had higher betaG levels in both gingiva and GCF than controls. Significant differences were observed among all groups when total GCF betaG activities were examined (P <0.05). However, the difference between AP and controls was not significant when concentration values were compared (P >0.05). The highest GCF betaG activity, with both expressions, was detected in EOP group. No absolute correlations between clinical parameters and betaG activity were observed, except for random correlations in the patient groups with mean total betaG activities. Also GCF/gingiva betaG levels and the 2 expressions did not show absolute correlations. CONCLUSIONS: The findings of the present study confirm the relationship between betaG activity and periodontal diseases. The differences in data concerning GCF total betaG activity and betaG concentration may suggest that they are not matching measures. Data presentation seems to be an important factor in GCF/enzyme profile studies. PMID- 10807128 TI - Osteoporosis and implant failure: an exploratory case-control study. AB - BACKGROUND: Osteoporosis has been suggested as a risk factor for implant failure, but data supporting such a link are limited. METHODS: A case-control study was designed to evaluate the association between osteoporosis and dental implant failure. Cases (n = 49) and controls (n = 49) were selected from a private practice. The following measures were collected for each patient: 1) peripheral dual-energy x-ray absorptiometry (pDEXA) bone measurements at the distal and proximal radius and ulna; 2) classification of bone quality and quantity at the time of implant placement; and 3) questionnaire data regarding potentially confounding variables. Generalized estimating equations were used to relate the likelihood of having at least one implant failure in an individual to osteoporosis measures. RESULTS: The results suggested that there was no association between pDEXA scores at the radius and ulna and the risk for implant failure. For every 1-unit increase in bone density as measured by pDEXA t-score at proximal and distal radius and ulna, the risk for a patient to lose at least one implant changed by +14% and -6%, respectively (odds ratio, 1.14; 95% confidence interval, 0.80-1.62 and odds ratio, 0.94; 95% confidence interval, 0.71-1.23). The simple visual assessment of local bone quality had a moderately sized relationship to implant failure. Implants placed in sites with thin cortical bone increased the chance for a patient to lose at least one implant by 130% when compared to implants placed in a thick cortical layer or compact bone (odds ratio, 2.3; 95% confidence interval, 1.0-5.4). CONCLUSIONS: This exploratory study suggests that a simple visual assessment of bone quality at the site of implant placement may be more informative regarding implant failure than pDEXA bone density measures obtained at peripheral bones. In order to determine whether peripheral osteoporosis measurements have a small or moderate association with implant loss, studies with larger sample sizes will be required. PMID- 10807129 TI - In vivo and in vitro natural frequency analysis of periodontal conditions: an innovative method. AB - BACKGROUND: When measuring periodontal disease, various types of equipment for making objective measurements of tooth mobility have been proposed. However, these devices and methodology are insufficient in terms of reliability. An innovative method using vibrational theories to assess the periodontal attachment level of natural teeth is presented in this study. METHODS: Modal testing technique, a non-destructive and time-saving method, was used for non-invasive and quantitative measurement of the natural frequencies of the upper central incisor in vivo and in vitro. A finite element model was established, and modal testing experiments were simulated to assess the relationship between bone level and teeth. RESULTS: The first dominant natural frequency of healthy human upper central incisors ranges from 710 Hz to 3,360 Hz, with an average of 1,701+/-679 Hz. Both in vitro experiment and finite element simulation showed that lowering of the attachment level causes a significant decrease in the natural frequencies, and a strong relationship (r = -0.99, P <0.01) was observed between the frequency and the bone level height. CONCLUSIONS: Our results suggested that natural frequency is an important parameter for assessing the periodontal condition and that the modal testing technique is a useful supplemental method for assessing the attachment conditions of the periodontium. PMID- 10807130 TI - Hepatocellular carcinoma metastatic to the oral mucosa: report of a case with multiple gingival localizations. AB - BACKGROUND: Metastases to the oral mucosa are rare, representing less than 1% of the tumors at this site. Most of these metastatic neoplasms originate in the lungs, kidneys, and liver. METHODS: The clinicopathologic features of an occult hepatocellular carcinoma, metastatic to the oral mucosa, are reported. The patient, a 70-year-old male, complained of 3 distinct polypoid, reddish lesions of the antero-inferior alveolar crest and both the right and left postero superior attached gingiva, without bone involvement. The lesions were excised, with the clinical diagnosis of multiple vascular tumors, formalin-fixed, paraffin embedded, cut and stained with hematoxylin and eosin. Consecutive sections were immunostained for alpha-1-antichymotrypsin, CEA, cytokeratins, EMA, hepatocyte antigen, PSA, S-100 protein, and thyroglobulin, using the alkaline phosphatase/anti-alkaline phosphatase technique. RESULTS: The morphologic features of the lesions were consistent with the diagnosis of carcinoma with trabecular and glandular patterns and bile secretion; furthermore, immunohistochemical reactivity for alpha-1-antichymotrypsin, cytokeratins, CEA, EMA, and hepatocyte antigen was demonstrated and the hepatic origin of the tumor was postulated. Ultrasonography demonstrated a liver mass, which was biopsied and treated by chemoembolization. While no further complications occurred in the oral mucosa, the patient died 8 months after the diagnosis for widespread diffusion of the tumor to the lungs and brain. CONCLUSIONS: This case emphasizes the need to include metastatic tumors in the differential diagnosis of atypical neoplasms of the oral mucosa and to evaluate the opportunity of surgical treatment in order to preserve the functions of the mouth, even if the prognosis of the primary tumors remains unfavorable. PMID- 10807131 TI - Burkitt's lymphoma presenting with jaw lesions. AB - We report an unusual case of Burkitt's lymphoma (BL) presenting with jaw lesions in a 14-year-old Chinese boy. The patient presented initially with mobile teeth in all 4 jaw quadrants, with corresponding radiographic detection of alveolar bone crest destruction and periapical bone resorption in the absence of clinically detectable jaw tumors. Moreover, radiographs taken only 17 days later showed clearly distinguishable signs of more extensive alveolar bone destruction compared with the initial radiographs. PMID- 10807132 TI - Cyclosporine A-induced gingival hyperplasia pemphigus vulgaris: literature review and report of a case. AB - Gingival hyperplasia appears in 8% to 85% of patients treated with cyclosporine. Most studies show an association between oral hygiene status and the prevalence and severity of this gingival overgrowth. Thus, besides attempting to substitute this drug with another whenever possible, treatment usually involves maintenance of strict oral hygiene coupled with scaling and root planing and removal of iatrogenic factors. Sometimes a second treatment phase involving periodontal surgery is necessary. Cyclosporine-induced gingival overgrowth has been mainly described in post-organ transplant patients. The present case describes, for the first time, a severe form of cyclosporine-induced gingival overgrowth arising in a 15 year-old male with pemphigus vulgaris. Periodontal treatment included oral hygiene and scaling and root planing under local anesthesia. There was a significant reduction in gingival enlargement, as well as a reduction in plaque levels and inflammation. Cessation of drug administration, combined with continuous periodontal treatment, brought further improvement. This successful conservative treatment of cyclosporine-induced gingival overgrowth in a pemphigus vulgaris patient suggests that early diagnosis and comprehensive treatment of these lesions may yield good response and reduce the need for periodontal surgery. PMID- 10807133 TI - Efficacy of periodontal therapy: statistical versus clinical significance. PMID- 10807134 TI - Diabetes and periodontal diseases. Committee on Research, Science and Therapy. American Academy of Periodontology. AB - This position paper on diabetes mellitus was prepared by the Research, Science and Therapy Committee of The American Academy of Periodontology. It is intended to: 1) update members of the dental profession on the diagnosis and medical management of patients with diabetes mellitus; 2) summarize current knowledge on the relation between diabetes mellitus and periodontal diseases; 3) provide an overview of factors in diabetic patients relevant to understanding the pathogenesis of periodontal diseases in these subjects; 4) outline special considerations associated with treatment of periodontal diseases in diabetic patients; and 5) discuss possible approaches to the management of diabetic emergencies in the dental office. PMID- 10807135 TI - Summary of the brush up on wellness symposium. San Antonio, Texas. September 24, 1999. PMID- 10807136 TI - Populist instead of professional. PMID- 10807138 TI - Screening for type II diabetes mellitus. PMID- 10807137 TI - Screening for hypothyroidism in adults: supporting data from two population studies. PMID- 10807139 TI - Participation in maternal serum screening following screen positive results in a previous pregnancy. AB - OBJECTIVE: To determine whether women who have had a positive serum screening result in one pregnancy have a lower rate of participation in screening in their next pregnancy. SETTING: The Women and Infants Hospital triple marker screening programme. METHODS: Pregnancy and screening information was collected from laboratory and hospital databases to compare subsequent screening participation in women who were screen negative and screen positive for risk of Down's syndrome (DS) or neural tube defect (NTD) pregnancy. RESULTS: In an age matched comparison, 108 women who had a previous screen positive result were significantly less likely than 108 women who were screen negative to participate in maternal serum screening in their next pregnancy. When examined according to type of screen positive result, the effect was significant for both those who were screen positive for DS and those who were screen positive for NTD. The degree of risk in screen positive women did not significantly affect their uptake of screening in the next pregnancy. CONCLUSIONS: Anxiety related to a screen positive result probably causes decreased participation in maternal serum screening in the next pregnancy. Reducing the screen positive rate in prenatal serum screening would alleviate maternal anxiety and would probably lead to more stable participation. PMID- 10807140 TI - Strategic options for antenatal screening for syphilis in the United Kingdom: a cost effectiveness analysis. AB - OBJECTIVE: Antenatal screening for syphilis is well established in the United Kingdom. The prevalence of syphilis is now very low, prompting the question as to whether this screening programme is still necessary. This paper aims at identifying possible screening strategy options for the programme and comparing their effectiveness and cost effectiveness. METHODS: The cost of the screening programme in the United Kingdom was estimated. This was based on the cost of screening tests, treatment, and follow up of infected women and their infants. This information was obtained from laboratories, antenatal clinics, and genitourinary medicine clinics. Epidemiological data from a survey of women treated for syphilis in pregnancy were analysed to identify groups at increased risk of syphilis. Strategic options for the screening programme were then identified. The effectiveness, number needed to treat, and cost effectiveness of these options were compared. RESULTS: Antenatal screening in the United Kingdom detected at least 40 pregnant women who need treatment for syphilis every year. This means that 18602 women are screened for every woman detected who needs treatment for syphilis. The marginal annual cost of this screening programme in the United Kingdom is 672366 pounds sterling. This is equivalent to 90p per woman screened, or 16670 pounds sterling to detect one woman who needs treatment for syphilis. The screening programme could be targeted geographically at pregnant women in the Thames regions. This option has the potential to save 482185 pounds sterling. Other strategic options are to target pregnant women in non-white ethnic groups, or those born outside the United Kingdom. These targeted options would each detect between 70% and 77% of women needing treatment for syphilis. These options could potentially save 592938 pounds sterling and 562691 pounds sterling respectively. CONCLUSIONS: Targeting or stopping the screening programme would save relatively little money. Although selectively screening groups by country of birth or by ethnic group could detect at least 70% of cases, this would be politically and practically difficult. Targeting by region would facts and the changing international epidemiology of syphilis lead us to recommend that the current universal antenatal screening for syphilis should continue. PMID- 10807141 TI - Implementation of service screening with mammography in Sweden: from pilot study to nationwide programme. AB - Establishment of mammography screening in Sweden has progressed logically from pilot study through clinical trials to service screening. Screening with mammography for early detection of breast cancer has been provided by all Sweden's 26 county councils since 1997. It took 23 years from the initial pilot study through clinical trials to the establishment of mammography service screening throughout Sweden. In the screening rounds completed by 1995-96, and provided by all but one county council, 1040000 women participated, corresponding to 81% of those invited. The national average recall rate was 2.2%, and consequently 23000 women were recalled for additional investigations. Eleven county councils invited women aged 40-74, six invited women aged 50-69, the remaining eight invited women between both these age intervals. Mammography outside screening programmes-clinical mammography-is available throughout Sweden. About 100000 women a year were referred for clinical mammography and about 50% of these were either younger or older than those invited for screening. A negative relation between the use of clinical mammography and participation in the screening programmes was noticed. PMID- 10807142 TI - A blind review and an informed review of interval breast cancer cases in the Limburg screening programme, the Netherlands. AB - OBJECTIVE: To investigate the proportion of interval breast cancers that could have been detected at the previous screening examination, and to gain more insight into the characteristics of these tumours. SETTING: Breast cancer screening programme in mid- and southern Limburg, the Netherlands. METHOD: Firstly, previous screening mammograms of 92 interval cancer cases were blindly reread by the radiologists from two different units as part of their daily screening workload. Secondly, a separate informed review was conducted, in which all of the cases (except for two) were subclassified into four categories: screening error; minimal signs; radiologically occult both at previous screening and at diagnosis; or radiologically occult at previous screening. Trends in recall rates and false and true positive rates were calculated to study potential changes during and after the blind review. RESULTS: In the blind review, 15% of the interval cancers were detected in both rereadings and 21% in one rereading. In the informed review, 25% were classified as a screening error and 24% as minimal signs present; 74% of the cases classified as a screening error had been recalled at least once in the blind review. Recall rates and false positive rates in daily screening practice increased significantly during and after the study period. CONCLUSIONS: About one third of the interval cancers could have been detected in the previous screening round. In order to diminish the number of avoidable interval cancers, review and classification of interval cancers is an important tool for continuing the education of screening radiologists. Therefore, further development of review training procedures is necessary. PMID- 10807143 TI - Improving the detection of cancer in the screening of mammograms. AB - OBJECTIVE: Examination of figures from screening at several different centres, and from the incidence of interval cancers in those programmes, suggests that a single radiologist reading by him/herself would miss 16 to 31% of cancers detectable in the mammograms. This study investigates how the accuracy of breast cancer screening could be improved. METHOD: Draw a comparison between screening for breast cancer and other inspection tasks that have been studied in the experimental psychological laboratory to suggest two low cost fixes that might improve the accuracy of breast cancer screening. RECOMMENDATIONS: (1) Radiologists should not work for more than half an hour at a time; (2) two radiologists double reading should inspect the same series of mammograms in inverse order; (3) there should be a clerical check to see whether the efficiency of detection decreases with time on task; and (4) a more sophisticated engineering of the screening procedure might be accomplished by assembling a library of mammograms for which the correct diagnosis is known. If these library mammograms were mixed randomly with those to be inspected in some suitable ratio, it would then be possible to provide the radiologist with immediate feedback on the accuracy of (some of) his/her diagnoses. From experimental studies of inspection tasks this should improve diagnostic accuracy. These recommendations, of course, require evaluation in field trials. PMID- 10807144 TI - Cost of quality management and information provision for screening: colorectal cancer screening. AB - OBJECTIVE: To estimate the costs of a quality management (QM) system as proposed by the Quality Management for Screening report for a future national colorectal cancer screening programme. METHODS: Estimates of the costs of the QM system, including the associated costs of education and training and information provision, were based on expert opinion, the existing literature, and the experience of the current National Health Service (NHS) breast cancer screening programme (BSP) and the NHS cervical cancer screening programme (CSP). RESULTS: The cost of a QM system to support a national colorectal cancer programme in the UK was estimated as approximately 3.8 million a year. Further annual costs related to QM will include 500000 for education and training and 200000 for information provision. Adding these additional costs to a previously published UK economic evaluation of colorectal cancer screening increases the cost-utility ratio to approximately 6500 per quality adjusted life year gained (over an eight year follow up period). CONCLUSIONS: Any new screening programme, or an existing one, must have QM to ensure that the quality of screening is high and to maintain the right balance between benefit and harm. The significant costs of such a QM system should be included in any economic evaluation of a screening programme. PMID- 10807145 TI - Screening for colorectal cancer by faecal occult blood test: comparison of immunochemical tests. AB - OBJECTIVE: To compare two immunochemical faecal occult blood tests based on reversed passive haemagglutination (RPHA) or latex agglutination (Hdia) in a population based screening setting. METHOD: Hdia was interpreted according to three positivity thresholds: 100, 150, or 200 ng of haemoglobin/mg of specimen solution. A total of 5844 subjects were recruited into the study, from 17432 invited subjects aged 50-70. RESULTS: Positivity rates were 3.3% for RPHA, Hdia100 3.5%, Hdia150 2.5%, Hdia200 2.0%. Among subjects complying with the diagnostic work up, colorectal cancer (CRC) was detected in 19 subjects (17 RPHA positive, 16 Hdia100 positive, 15 Hdia150 positive, 14 Hdia200 positive) and high risk adenoma/s in 41 subjects (28 RPHA positive, 32 Hdia100 positive, 29 Hdia150 positive, 25 Hdia200 positive). The prevalence of screen positive CRC in the population was for RPHA 2.9 per thousand, Hdia100 2.7 per thousand, Hdia150 2.6 per thousand, Hdia200 2.4 per thousand. The prevalence of screen positive high risk adenomas in the population was for RPHA 4.8 per thousand, Hdia100 5.5 per thousand, Hdia150 5.0 per thousand, Hdia200 4.3 per thousand. CONCLUSION: Hdia100 was as sensitive as RPHA for cancer and high risk adenomas. As Hdia is less technically complex than RPHA, it is a valid alternative to the latter, provided that full automation of the development procedure is available. Increasing the positivity threshold of Hdia up to 150 or 200 ng of haemoglobin/mg of specimen solution is not advisable as the increase in specificity is too small to justify the corresponding decrease in the detection of screen positive cancers in the population. PMID- 10807146 TI - Acceptability of flexible sigmoidoscopy screening in older adults in the United Kingdom. AB - OBJECTIVES: To assess the acceptability of bowel cancer screening using flexible sigmoidoscopy (FS). SETTING: Adults aged 55 to 64 recruited from general practices in Welwyn Garden City and Leicester, which were the pilot and start up centres of a multicentred randomised controlled trial of FS screening (the ICRF/MRC Flexible Sigmoidoscopy Screening Trial). METHODS: Screenees (n=4422) were sent a three month follow up questionnaire that included measures of satisfaction with information given before the test, facilities at the test unit, attitudes of the staff, and explanation of the results. Measures of pain, embarrassment, feelings of being "in control" during the test, willingness to encourage others to have the test, and gladness to have participated were also included. In addition, semistructured telephone interviews were conducted with 60 screenees, stratified by screening outcome and gender. RESULTS: The follow up questionnaire was completed by 94% of screenees. Responses indicated a high level of satisfaction with the procedure: 99% were satisfied with the information given before the test, the facilities, the attitudes of the staff, and the explanation of their results; 91% reported only mild or no pain; 97% reported little or no embarrassment; and 99% were glad they had the test. Satisfaction ratings varied little by gender or outcome group. The quantitative results were reinforced by the qualitative data, which also revealed high acceptability. CONCLUSION: In the context of a clinical trial with dedicated trial staff, FS is a well tolerated procedure. There are high levels of satisfaction with service provision and positive attitudes towards the programme. PMID- 10807147 TI - Glucose-6-phosphate dehydrogenase deficiency neonatal screening: preliminary evidence that a high percentage of partially deficient female neonates are missed during routine screening. AB - OBJECTIVES: To provide preliminary evidence that the currently employed semiquantitative method of screening for glucose-6-phosphate dehydrogenase (G6PD) deficiency can only detect infants who are totally deficient for G6PD and misses all cases of partial G6PD deficiency. SETTING: General population: 2150 randomly selected blood samples from the Blood Donation Department, Speliopouleion General Hospital, Athens, Greece. Neonate population: 2000 samples from neonates (50% male; 50% female) in maternity hospitals in the greater Athens area. High risk population: a total of 545 individuals from 133 families in the Athens area, the minimum acceptance criteria being the parents and any brother or sister. METHOD: Blood specimens from neonates or adults were collected and either spotted and dried on special filter paper (Schleicher and Schull 2992, Darmstadt, Germany) or used in tubes after being heparinised. For the quantitative evaluation of G6PD enzyme activity, the Quantase G6PD screening kit (Quantase Limited, Perth, UK) was used. Quantase G6PD controls (Quantase Limited) were used at three levels of G6PD. These controls are rated at 24, 30, and 37 degrees C. Alternatively, we used the Sigma G6PDH controls (Sigma Chemical Company, St Louis, USA) which are rated at 30 and 37 degrees C. The assay was performed according to the instructions included in the kit with the modification for haemoglobin normalisation. RESULTS: General population: 36 females who were classified as having normal enzymatic activity with the semiquantitative test, were classified as partially deficient with the quantitative test. Neonate population: using the quantitative test, the percentage of G6PD deficient neonates in this population was 5.5%, compared with 3.17% reported in routine screening using the semiquantitative method. High risk population: the quantitative method detected 28 cases of total or partial G6PD deficiency in sisters of males with known total deficiency. The semiquantitative CONCLUSIONS: A considerable amount of partially G6PD deficient female neonates (heterozygotes) are undetected and classified as having normal enzymatic activity using the semiquantitative method, which uses a cut off of 2.1 U/g haemoglobin (Hb). The use of a fully quantitative G6PD screening kit is proposed, employing the automated haemoglobin normalisation and a cut off of 6.4 U/g Hb. Any neonate with an activity below this mark should be regarded as G6PD deficient, and all preventive measures should be taken. PMID- 10807148 TI - Reporting of screening results. PMID- 10807149 TI - Introduction to the special issue on stress and substance use. PMID- 10807150 TI - The variable effects of stress on alcohol use from adolescence to early adulthood. AB - Despite evidence of a strong association between stress and level of drinking in adolescent populations, the role of stress in accounting for changes in drinking behavior throughout the adolescent years is unclear. This study uses a linear growth curve analysis to examine the determinants of within-individual changes in drinking frequency and binge drinking across five waves of data from a community sample of adolescents who were followed into young adulthood. Predictors of drinking include: stressful life events, parental and peer social support, and parental and peer relationship problems. Findings indicate significant effects of stressful life events and parental support and conflict on both the frequency and intensity of alcohol use. Although age-related changes in these variables coincide with changes in drinking behavior, they do not account for drinking variability over this period. Results from conditional models demonstrate that the impact of the stress is contingent on age, and that the strong associations between drinking and stress evidenced during the high school years weaken considerably as individuals move into their late teens and early twenties. Discussion centers on the complex motivations for and facilitators of drinking as young people mature and change environments over the adolescent years. PMID- 10807151 TI - Alcohol problems among a community sample: longitudinal influences of stress, coping, and gender. AB - This study examined the relationships among stress, coping methods, and alcohol use associated problems within a longitudinal, community sample of males and females who were followed from adolescence through young adulthood. While fewer females than males were categorized as "dependent users," these females were more debilitated in terms of the number of alcohol-related problems experienced from age 15 through age 25. Measures of both chronic and proximal levels of personal stress and negative coping styles were most helpful in explaining problems with alcohol use. Distal measures of stress and coping, while perhaps serving as mediators or moderators of other important constructs, did not appear to have a direct effect on the outcome measure. PMID- 10807152 TI - A growth curve analysis of stress and adolescent drug use. AB - The purpose of this paper is to describe and examine one pathway by which adolescent drug use increases during early and mid-adolescence. It draws upon recent research on adolescent stress, drug use, and family processes to describe an important stage of the life course. A chief principle underlying the proposed pathway is that the cumulative effect of stressful life experiences over time can lead to a steeper escalation of drug use in adolescence. Furthermore, based on previous stress research, we propose that this effect may be moderated by factors such as sex, income, family attachment, self-esteem, and mastery. Using 4 years of panel data from the Family Health Study (n = 651 adolescents ages 11-14 during Year 1), we estimate a hierarchical growth curve model that examines the time varying effects of stressful life events and peer relations on drug use. The results indicate that experiencing a high number of life events over time is related to a significant "growth" of drug use, even after controlling for "growth" due to age or peer relations. In addition, this relationship is moderated by family attachment; high levels of attachment serve to diminish this growth significantly. PMID- 10807153 TI - One-year prospective prediction of drug use from stress-related variables. AB - Six stress-related variables, gender, age, and ethnicity were investigated as concurrent and prospective predictors of three types of drug use (cigarettes, alcohol, and illicit drug use) among 875 "high risk" adolescents. The stress related variables were socioeconomic status, "missing" one's parent(s), family conflict, victimization, perceived stress, and stress-drug beliefs. In general, concurrent regression models indicated that those who were lower in socioeconomic status, held stress-drug beliefs favorable toward drug use, and who had been victimized in the last year were more likely to be cigarette, alcohol, or illicit drug users. Prospective regression models indicated that those who had used drugs at baseline and had been victimized in the last year were relatively likely to use drugs the next year. Significant predictors in the multivariable models accounted for between 56 and 85% of those subjects who were above the median on later drug use. Victimization is apparently a relatively important source of stress in the prediction of future drug use. Drug-use intervention implications of these results include the need to provide supportive services to those who have been victims of violent attacks on their person or property. PMID- 10807154 TI - Work stressors, drinking with colleagues after work, and job satisfaction among white-collar workers in Japan. AB - Although previous studies have examined the buffering effects of social support and coping style on the relationship between stress and alcohol consumption, they have typically relied on analysis of variance (ANOVA) or regression analysis. In addition, few studies have examined the potential stress-buffering effects of drinking with coworkers after work on the relationship between job stress and job dissatisfaction. In the present study, using a signal detection analysis, we evaluated the interactions of drinking with coworkers after work and work stressor variables among Japanese white-collar workers (n = 397) in 1997. The analysis was performed for two groups of subjects divided based on their status in the company. This was necessary because in Japan the obligations to drink socially increase with one's rising status in the company. In both the "staff members and lower-level managers" and "middle-level and higher-level managers' groups, an interaction between work-stressor variables and drinking variables was observed. The findings imply that drinking with coworkers after work ameliorated the sense of job dissatisfaction, but only among those subjects who already had lower levels of work stressors. For subjects with high levels of work stressors, attitudes toward drinking with coworkers were unrelated to job satisfaction levels. PMID- 10807155 TI - A bibliographic essay: the relationship between stress and substance use. AB - The purpose of this review is to outline and critique studies that have examined the link between stress and substance use. Studies are categorized according to the age of the sample and the type of substance use measure considered. The research is mostly limited to studies that operationalize stress as stressful life events, daily hassles, or subjective stress. This paper concludes that there is a robust relationship between stress and substance use. However, there is a clear need for general population studies to address age, gender, and cultural differences to better assess the specificity and complexity of the stress/substance use relationship. PMID- 10807156 TI - Review: Biology and relevance of C-reactive protein in cardiovascular and renal disease. AB - C-reactive protein (CRP) is a member of the pentraxin family of proteins, which are characterised by a cyclic pentameric structure and radial symmetry. The five identical 24-kDa protomers consist of 206 amino acids, and are noncovalently linked. CRP binds to a range of substances such as phosphocholine, fibronectin, chromatin, histones, and ribonucleoprotein in a calcium-dependent manner. It is a ligand for specific receptors on phagocytic leukocytes, mediates activation reactions on monocytes and macrophages, and activates complement. Plasma CRP is the classical acute-phase protein, increasing 1,000-fold in response to infection, ischemia, trauma, burns, and inflammatory conditions. A growing number of studies suggest that CRP is an independent risk factor for atherosclerotic vascular disease. Plasma CRP concentrations in the highest quartile are associated, depending on the subject group, with 1.5- to 7-fold increases in relative risk. In the high-risk endstage renal failure population, a raised CRP is associated with up to 5.5-fold increased relative risk of CVD and 4.6-fold increased relative risk of death. This review examines the relationships between CRP, cardiovascular disease, and mortality, with special reference to renal disease. PMID- 10807157 TI - Review: Free radicals, antioxidants, and the immune system. AB - Oxygen-derived free radicals are important in both natural and acquired immunity. Neutrophil and macrophage phagocytosis stimulates various cellular processes including the "respiratory burst" whereby increased cellular oxygen uptake results in the production of the potent oxidant bactericidal agents, hypochlorous acid and hydroxyl radical. In addition, nitric oxide, a gaseous radical produced by macrophages, reacts with superoxide to form peroxynitrite, also a potent bactericidal agent. Conversely, oxidative stress may be detrimental in acquired immunity by activation of nuclear factor kappa B, which governs gene expression involving various cytokines, chemokines, and cell adhesion molecules, among others. However, antioxidant supplementation essentially reverses several age associated immune deficiencies, resulting in increased levels of interleukin-2, elevated numbers of total lymphocytes and T-cell subsets, enhanced mitogen responsiveness, increased killer cell activity, augmented antibody response to antigen stimulation, decreased lipid peroxidation, and decreased prostaglandin synthesis. PMID- 10807158 TI - Commentary: Human mitochondrial cytopathies. AB - Mitochondria provide energy (ATP) for all eukaryotic cells except mature erythrocytes and keratinocytes. They are abundant in cells that expend much energy, such as muscle, exocrine pancreas, nervous system, and heart cells, and motile sperm. Many mitochondrial enzymes are encoded by nuclear DNA and imported into the mitochondria. Like bacteria, mitochondria possess their own DNA and ribosomes. They are fueled by fatty acids and pyruvate, and through acetyl-coA enzyme can use fats, carbohydrates, and proteins as energy sources, producing ATP for cells. A high index of suspicion for mitochondrial mutations enables clinicians to recognize these unusual and rare disorders and provide proper genetic counseling. Mitochondrial cytopathies include a diverse group of diseases, affecting many organs, especially skeletal muscle and central nervous system, and are associated with abnormal mitochondria in skeletal muscle known as ragged red fibers. Mitochondrial DNA mutations are detectable in peripheral blood. PMID- 10807159 TI - Commentary: Iron metabolism in hepatitis C infection. AB - Hepatitis C virus (HCV) infections have started to decline, but up to 10,000 deaths each year are the consequence of chronic liver disease, following the infection. Laboratory testing identifies HCV-infected individuals using positive recombinant immunoblot assays to detect the presence of the antibody; the diagnosis is confirmed by detecting HCV RNA in serum. HCV-infected patients who have large accumulations of hepatic iron have not responded well to interferon therapy, compared to patients with normal hepatic iron stores. Physicians who treat patients infected with HCV should be aware of the detrimental effect of excess liver iron on interferon therapy. The degree of hepatic iron overload should be assessed and the reason for the excess iron should be investigated. Phlebotomy is the most practical method for iron removal and is well tolerated by patients with HCV infection. PMID- 10807160 TI - Discordant results of CK-MB and troponin I measurements: a review of 14 cases. AB - In the course of a clinical comparison involving 204 parallel total creatine kinase (CK), creatine kinase-MB isoenzyme (CK-MB), and cardiac troponin I (cTnI) measurements, 12 patients were identified in whom cTnI was elevated while total CK was normal, as well as 2 patients in whom CK-MB was elevated while cTnI was normal. CK-MB relative index was elevated in 6 of the twelve cTnI-positive patients with normal total CK; only 2 of these patients had a discharge diagnosis of acute myocardial infarction (AMI). All of the 12 patients in this group had medical conditions that are associated with greater risk for acute cardiac events. Both patients with normal cTnI but elevated total CK and CK-MB index had chronic renal insufficiency; one of these patients had a positive stress test and a diagnosis of AMI. The other cTnI-negative patient died 2 days after admission, and autopsy revealed evidence of ischemic changes, but not acute infarction. Significant differences were apparent between traditional CK-MB results and cTnI measurements. Using total CK elevation as a prerequisite for subsequent CK-MB measurement may limit the clinical sensitivity of this enzyme marker for detecting subacute ischemic damage to the myocardium. Elevated total CK and CK-MB isoenzyme without corresponding elevations in cTnI, on the other hand, may reflect changes in enzyme elimination kinetics due to renal failure, or cross reactivity of the cTnI assay with non-cardiac antigens. PMID- 10807161 TI - Elevated serum chromogranin A is detectable in patients with carcinomas at advanced disease stages. AB - Chromogranin A (CgA), a marker of neuroendocrine cells and an indicator for neuroendocrine differentiation, is associated with a poor prognosis when detected in tumor tissue, based on immunohistochemical techniques. We sought to determine whether it is possible to detect elevated serum CgA in patients with commonly occurring carcinomas of non-neuroendocrine origin. CgA was measured in both random and serial serum specimens, using a serum CgA assay developed in our laboratory. Elevated levels of serum CgA were detected in patients with carcinoma of the prostate, breast, ovary, pancreas, and colon. Serum CgA levels in patients with all types of carcinoma appeared to parallel the changes of serum dominant tumor markers and were found in sera containing highly elevated tumor markers. Based on these preliminary findings, perhaps we should monitor CgA, in addition to the routinely used tumor markers, during the treatment of patients with carcinomas to determine if CgA is useful as a prognostic marker in carcinomas other than prostatic cancer. PMID- 10807162 TI - Autoantibodies to p53 in sera of patients with autoimmune thyroid disease. AB - Mutations in the tumor suppressor gene, p53, lead to intracellular accumulation of abnormal p53 protein and are associated with p53 autoantibodies. p53 also accumulates in autoimmune diseases and Hashimoto's thyroiditis, but it is unknown if p53 autoantibodies occur in the latter. We measured p53 autoantibodies in the sera of 93 patients with thyroid disease and 19 patients without thyroid disease. Anti-p53 antibodies were detected in the sera from 4.2% (2/48) of patients with autoimmune thyroid disease, including one patient with Hashimoto's thyroiditis (3.7%, 1/27) and one with Graves' disease (4.8%, 1/21). A third patient with pseudohypoparathyroidism, but without thyroid disease, was also positive (1/19; 5.2%). None of 19 patients with differentiated thyroid cancer had anti-p53 antibodies. We conclude that anti-p53 antibodies can be detected in the sera from approximately 4% of patients with autoimmune thyroid disease. This finding suggests that increased DNA damage and apoptosis may be associated with autoimmune thyroid disease. PMID- 10807163 TI - Multicenter analytical evaluation of an automated immunoassay for total plasma homocysteine. AB - A fully automated immunoassay for total plasma homocysteine assay was evaluated at four centers. To measure total homocysteine, oxidized forms of homocysteine in serum and plasma were reduced by dithiothreitol and assayed by a competitive fluorescence polarization technique. The assay had within-run precision from 0.9 to 3.0% and total precision from 2.8 to 4.1% for control materials with homocysteine concentrations of approximately 7, 12.5, and 25 micromol/L, a sensitivity of 0.35 micromol/L, good parallelism upon dilution, and analytical recovery ranging from 97.4 to 103.8%. The immunoassay correlated with four different HPLC assays for homocysteine, yielding a slope of 0.98, an intercept of -0.19 micromol/L, and a correlation coefficient of 0.966 for 440 paired samples. The reference range, determined with plasma samples from 609 males and 600 females, yielded a mean of 9.17+/-2.86 micromol/L, with a central 95% range of 4.78-15.43 micromol/L. The immunoassay is a suitable alternative to HPLC and may be useful in screening persons with high risk of coronary artery disease. PMID- 10807164 TI - Chromatographic measurements of hemoglobin A2 in blood samples that contain sickle hemoglobin. AB - In the sickle cell syndromes, Hb A2 measurements aid in the differential diagnosis of sickle cell anemia from sickle-beta-thalassemia. The purpose of this study is to assess the Hb A2 levels in samples containing sickle hemoglobin (Hb S) by the use of an automated high performance liquid chromatography system (HPLC Variant beta-thalassemia Short Program). The blood samples analyzed were from individuals of African descent living in the state of Tennessee who had either sickle cell trait (Hb AS), sickle cell disease (Hb SS), or sickle cell-hemoglobin C disease (Hb SC). Interestingly, the Hb A2 levels determined by HPLC were found elevated in samples containing Hb S. The Hb A2 mean in Hb AS samples (n=146) is 4.09% (SD +/- 0.42, range 2.20 to 5.20%); in Hb SS samples (n=33) it is 3.90% (SD +/- 1.08, range 0.60 to 5.90%); and in Hb SC samples (n=27) it is 4.46% (SD +/- 0.70, range 2.30 to 5.91%). The Hb A2 mean by HPLC in normal individuals (Hb AA, n=70) is 2.57% (SD +/- 0.25, range 2.1 to 3.0%), and the Hb A2 range in beta thalassemia carriers is 4 to 9%. Our results show that the Hb A2 levels in Hb S containing samples partially overlap with those expected from beta-thalassemia carriers. The hemoglobinopathy laboratory should be aware of this apparent elevation in Hb A2 levels determined by HPLC in individuals carrying Hb S. Other factors, such as family history and clinical symptoms, should be taken into account before a diagnosis of sickle cell trait, sickle-beta-thalassemia, or sickle cell anemia is made. PMID- 10807165 TI - Hemolysis during leukocyte-reduction filtration of stored red blood cells. AB - Hemolysis has been reported in red blood cells (RBCs) that have undergone leukocyte-reduction filtration. This study investigated whether the age of RBCs or the filter type affected hemolysis. One hundred eighty units of RBCs (adenine saline added) were leukocyte-reduced by filtration. At each of the 6 weeks of shelf life, 10 units were filtered with the "BPF4" filter, 10 units with the "Purecell RCQ" filter, and 10 units with the "Sepacell" filter. Filtration was performed with strict adherence to the manufacturers' directions. Pre- and post filtration samples were assayed for plasma hemoglobin by measuring the plasma absorbances at 578 nm and 562 nm. The increase of plasma hemoglobin concentration following filtration was significantly greater (p < 0.05) in older units, compared to fresher units, when the Sepacell and BPF4 filters were used. For example, the increase of plasma hemoglobin at week 6 (83.47 mg/dl:Sepacell, 128.93 mg/dl BPF4) was significantly greater than at week 1 (7.07 mg/dl Sepacell, 4.77 mg/dl BPF4) (Sepacell: p=0.008; BPF4: p=0.006). For units stored 1, 2, 4, 5, or 6 weeks, the increase of plasma hemoglobin concentration post-filtration was significantly greater with the BPF4 filter, compared to the Purecell RCQ filter (p <0.045); for units stored 5 weeks, the increase in plasma hemoglobin concentration post-filtration was significantly greater with the BPF4 filter compared to the Sepacell filter (p = 0.009). Mean filtration times were significantly longer in older units compared to fresh units. This study shows that increased storage duration of RBCs (adenine-saline added) is attended by greater hemolysis during leukocyte-reduction filtration and by prolongation of the filtration time. In addition, the amount of hemolysis may be influenced by the type of filter. PMID- 10807166 TI - Differential expression of chemokines in a mouse model of wound healing. AB - Macrophages have a multifaceted role in wound healing. While their initial activity may be in the degradation and elimination of damaged tissue, macrophages also produce and secrete a variety of mediators that can participate in the repair process as well. To perform these functions, macrophages must be recruited to a wound site. Our purpose was to examine the temporal and spatial expression of macrophage chemoattracting cytokines (chemokines) at a surgical wound site. A surgical wound was prepared on the dorsal aspect of B6AF1/J mice. Biopsies were obtained from the wound and a comparable nonwounded area between 6 and 72 hr after wounding. The presence or absence of various chemokine mRNAs was detected by the reverse transcriptase-polymerase chain reaction (RT-PCR). Immunohistochemical staining and in situ RT-PCR determined localization of cells producing chemokines. In wounded tissue, both macrophage chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein-1 (MIP-1) were detected; however, the time of expression differed for each molecule. MCP-1 mRNA was detected at 6 hr after wounding, with decreased expression at subsequent time periods. In contrast, MIP-1 messages were not observed until 24 hr after wounding, and steadily increased thereafter. MCP-1 and MIP-1 mRNA and protein were localized predominantly in keratinocytes. The rapid and strong expression of MCP-1 and MIP 1 messages within the wound site suggests a pivotal role for these chemokines in the repair process. The differences in appearance and level of expression over time, however, suggest distinctive functions for each chemokine and indicate that the local milieu, rather than a single cytokine, influences macrophage recruitment and/or activation. PMID- 10807167 TI - Antioxidant properties of pyrrolidine dithiocarbamate and its protection against Cr(VI)-induced DNA strand breakage. AB - Pyrrolidine dithiocarbamate (PDTC) is considered an antioxidant and is frequently used to study the role of free radical reactions in various biological processes and against free radical-induced cellular injuries. However, its antioxidant properties are not characterized. In this study, electron spin resonance (ESR) was used to investigate the antioxidant potential of PDTC with hydroxyl radical (*OH) and superoxide anion radicals (O2*-). The Fenton reaction [Fe(II) + H2O2 - > Fe(II) + *OH + OH-)] and xanthine and xanthine oxidase were used as sources of *OH and O2*- radicals, respectively. The results show that PDTC effectively scavenges *OH radicals with a reaction rate constant of approximately 2.73 x 10(10) M(-1) s(-1), which is comparable to other efficient *OH radical scavengers, such as ascorbate and glutathione. PDTC is also able to scavenge O2*- radicals. Through its antioxidant properties, PDTC protects against Cr(VI) induced DNA strand breakage. PMID- 10807168 TI - "Almost delocalized" intervalence compounds AB - Interest is high in +M-bridge-M systems that have very low barriers to intramolecular electron transfer giving M-bridge-M+ ("almost delocalized" systems). Hush showed how to evaluate the electronic coupling across the bridge (H) from the M-to-M charge-transfer optical absorption band, but did not point out that his classical model causes the extinction coefficient to suddenly drop to zero at a photon energy of 2 H. Ignoring this band cutoff leads to a low estimation of H. PMID- 10807169 TI - Different approaching directions of sigma and pi nucleophiles to the sulfur atom of thiiranium and thiirenium ions PMID- 10807170 TI - Anisotropic orientation of horseradish peroxidase by reconstitution on a thiol modified gold electrode. AB - Horseradish peroxidase (HRP) was reconstituted on the surface of a gold electrode that was modified first with a hemin-carbon-chain-thiol derivative followed by addition of the apo protein to the contacting solution. To facilitate the reconstitution of the holo enzyme, the hemin needs to be immobilised on a carbon chain spacer arm. To achieve this, an immobilisation protocol was developed that is based on the initial formation of a mixed self-assembled monolayer on the gold surface consisting of 3-carboxypropyl disulphide and an activated disulphide (3,3'-dithiodipropionic acid di-(N-succinimidyl ester)) followed by binding of a diaminoalkane to the activated disulphide. The hemin was then coupled to the second amino group of the diaminoalkane by means of a carbodiimide coupling reagent. Finally, the enzyme was reconstituted on the hemin-modified surface by immersion of the electrode in a solution containing apo-HRP. The advantage of this method is that the length of the spacer arm can be changed easily, because diaminoalkanes of different chain lengths are available. The electrochemistry of the hemin and the reconstituted HRP electrodes was studied by means of cyclic voltammetry and differential-pulse voltammetry. The catalytic ability for reduction of hydrogen peroxide was investigated for both direct and mediated electrochemistry with a soluble electron donor (ortho-phenylenediamine). PMID- 10807171 TI - Distorted bcc indium cubes as structural motifs in Ca AB - The title compounds were prepared from the elements by reactions in water-cooled glassy carbon crucibles under an argon atmosphere in a high-frequency furnace. CaPdIn4 crystallizes with the YNiAl4-type structure: Cmcm, a=446.7(3), b=1665(1), c=754.3(5) pm, wR2=0.0465 with 646 F2 values and 24 variables. The structure is built up from a complex three-dimensional [PdIn4] polyanion in which the calcium atoms occupy distorted pentagonal tubes formed by indium and palladium atoms. CaRhIn4 and CaIrIn4 adopt the LaCoAl4-type structure: Pmma, a=867.6(1), b=422.91(8), c=745.2(1) pm, wR2=0.0583 with 468 F2 values and 24 variables for CaRhIn4; a=869.5(1), b=424.11(6), c=746.4(1) pm, wR2= 0.0614 471 F2 values with 24 variables for CaIrIn4. This structure type, too, has a three-dimensional [RhIn4] polyanion which is related to the structure of binary RhIn3. The calcium atoms fill distorted pentagonal prismatic channels formed by indium atoms. Semi empirical band structure calculations for Ca-RhIn4 and CaPdIn4 reveal strongly bonding In-In, Rh-In and Pd-In interactions but weaker Ca-Rh, Ca-Pd and Ca-In interactions. CaRhIn4 and Ca-PdIn4 are compared with other indium-rich compounds such as YCoIn5 and Y2CoIn8, and with elemental indium. Common structural motifs of the indium-rich compounds are distorted bcc-like indium cubes. PMID- 10807172 TI - Photophysical and electrochemical characterisation of the interactions between components in neutral pi-associated AB - The electrochemical and photophysical properties of a variety of neutral pi associated [2]catenanes have been explored by using cyclic voltammetry, absorption and luminescence spectrophotometry and the measurement of exited-state lifetimes. Several trends that could be correlated with interactions between the mechanically linked components of the structures were revealed from comparative study of catenane precursors, model compounds and the [2]catenanes themselves. Throughout, emphasis is placed on pair-wise comparisons between systems which differ in a single structural feature. Substitution in a catenane of a pyromellitic diimide unit by a naphthalene diimide yields more readily reduced derivatives, whose absorption spectra reveal charge transfer within the catenane to be a lower energy process of reduced intensity. Conversion of the butadiyne links within the diimide macrocycle of the catenanes to saturated chains results in an increase in both the energy and intensity of their charge-transfer bands; electrochemically these derivatives are all harder to reduce than the parent systems. Replacement of one of the electron-donating components of the catenanes with a less effective aromatic donor bearing a carboxy group also decreases the energy and intensity of the charge-transfer feature and is accompanied by a slightly more ready reduction. A sequence of reduction and translational events is proposed to explain the intriguing electrochemical behaviour of a catenane that contains one pyromellitic and one naphthalene diimide. For some systems the photophysical and electrochemical techniques, whilst exploring distinct physical phenomena, are shown to be in good agreement by comparison of shifts of electrochemical reduction waves with those of charge-transfer absorption features. PMID- 10807174 TI - Hydroxo hydrido complexes of iron and cobalt (Sn-Fe-Sn, Sn-Co-Sn): probing agostic Sn...H-M interactions in solution and in the solid state AB - Bis(toluene)iron 9 reacts with Lappert's stannylene [Sn[CH(SiMe3)2]2] (4) to form the paramagnetic bis-stannylene complex [[(eta6-toluene)Fe-Sn-[CH(SiMe3)2]2]2] (10). Compound 10 reacts with H2O to form the hydroxo hydrido complex [(eta6 C7H8)(mu-OH)(H)-Fe-[Sn[CH(SiMe3)2]2]2] (12) in high yield; its solid-state structure has been elucidated by X-ray and neutron diffraction analysis. In agreement with the 1H NMR results, 12 contains a hydridic ligand whose exact coordination geometry could be determined by neutron diffraction. The 1H and 119Sn NMR analysis of 12 suggested a multicenter Sn/Sn/H/Fe bonding interaction in solution, based on significantly large values of J(Sn,H,Fe) = 640+/-30 Hz and J(119Sn,119Sn) = 4340+/-100 Hz. In solution, complex 12 exists as two diastereomers in a ratio of about 2:1. Neutron diffraction analysis has characterized 12 as a classical metal hydride complex with very little Sn...H interaction and a typical Fe-H single bond (1.575(8) A). This conclusion is based on the fact that the values of the Sn...H contact distances (2.482(9) and 2.499(9) A) are not consistent with strong Fe-H...Sn interactions. This finding is discussed in relation to other compounds containing M-H...Sn units with and without strong three-center interactions. The neutron diffraction analysis of 12 represents the first determination of a Sn-H atomic distance employing this analytical technique. The cobalt analogues [(eta5-Cp)(mu-OH)(H)Co [Sn[CH(SiMe3)2]2]2] (15) and [(eta5-Cp)(OD)(D)Co-[Sn[CH-(SiMe3)2]2]2] [D2]15, which are isolobal with 12, were prepared by the reaction of [(eta5-Cp)Co Sn[CH(SiMe3)2]2] (14) with H2O and D2O, respectively. The magnitude of J(Sn,H) (539 Hz) in 15 is in the same range as that found for 12. The molecular structure of 15 has been determined by X-ray diffraction which reveals it to be isostructural with 12. The coordination geometries of the Co(Fe)-Sn1-O-Sn2 arrangements in 12 and 15 are fully planar within experimental error. Compounds 10 and 15 are rare examples of fully characterized complexes obtained as primary products from water activation reactions. PMID- 10807173 TI - Stabilized rhodium(0) nanoparticles: a reusable hydrogenation catalyst for arene derivatives in a biphasic water-liquid system. AB - A colloidal system based on an aqueous suspension of rhodium(o) nanoparticles proved to be an efficient catalyst for the hydrogenation of arene derivatives under biphasic conditions. The rhodium nanoparticles (2-2.5 nm) were synthesized by the reduction of RhCl3 x 3H2O with sodium borohydride and were stabilized by highly water-soluble N-alkyl-N-(2-hydroxyethyl)ammonium salts (HEA-Cn). These surfactant molecules were characterized by measurements of the surface tension and the aqueous dispersions with rhodium were observed by transmission electron cryomicroscopy. The catalytic system is efficient under ultramild conditions, namely room temperature and 1 atm H2 pressure. The aqueous phase which contains the protected rhodium(0) colloids can be reused without significant loss of activity. The microheterogeneous behavior of this catalytic system was confirmed on a mercury poisoning experiment. PMID- 10807175 TI - Heteropolynuclear complexes with the ligand Ph2PCH2SPh: theoretical evidence for metallophilic Au-M attractions AB - Addition of two equivalents of diphenylthiomethylphosphine (PPh2-CH2SPh) to the starting materials [Au(tht)2]A (tht = tetrahydrothiophene), AgCF3SO3, or [Cu(CH3CN)4]CF3SO3 produces the mononuclear derivatives [M(PPh2CH2SPh)2]A (M = Au, A = CF3SO3 (1a); M = Au, A = ClO4 (1b); M = Ag, A = CF3SO3 (4); M = Cu, A = CF3SO3 (5)) which are able to form the heterodinuclear complexes [AuM'(PPh2CH2SPh)2](CF3SO3)2 (M' = Ag (2), Cu (3)) with a P-Au-P environment. If the starting gold complex is [Au(C6F5)(tht)], reaction with the phosphine produces [Au(C6F5)-(PPh2CH2SPh)] (6) from which, by reaction with AgCF3SO3 or [Cu(CH3CN)4]CF3SO3, the "snake"-type linear complexes [Au2M(C6F5)2 (PPh2CH2SPh)2]CF3SO3 (M = Ag (7), Cu (8)) are obtained. If the silver starting complex is AgCF3CO2, reaction in a 1:1 ratio gives the tetranuclear complex [Au2Ag2(C6F5)2(PPh2CH2SPh)2-(CF3CO2)2] (9). When the molar ratio is 1:2 the trinuclear complex [AuAg2(C6F5) (CF3CO2)2(PPh2CH2SPh)] (10) is obtained. According to ab initio calculations, the presence of only one gold atom is enough to induce metallophilic attractions in the group congeners, and this effect can be modulated depending on the gold ligand. PMID- 10807177 TI - Enantioselective total synthesis of Wieland-Gumlich aldehyde and (-)-strychnine AB - A total synthesis of (-)-strychnine in 15 steps from 1,3-cyclohexanedione in 0.15% overall yield is described. The sequence followed in the assembling of rings is: E-->AE [2-(2-nitrophenyl)-1,3-cyclohexanedione]-->ACE (3a aryloctahydroindol-4-one)-->ACDE (arylazatricyclic core)-->ABCDE (strychnan skeleton)-->ABCDEF (Wieland-Gumlich aldehyde)-->ABCDEFG (strychnine). The key steps of the synthesis are the enantioselective construction of the 3a-(2 nitrophenyl)-octahydroindol-4-one ring system and the closure of the piperidine ring by a reductive Heck cyclization to generate the pivotal intermediate (-)-14. In contrast, a Lewis acid promoted a-alkoxypropargylic silane-enone cyclization did not lead to synthetically useful azatricyclic ACDE intermediates. The introduction of C-17 and the closure of the indoline ring by reductive amination of the alpha-(2-nitrophenyl) ketone moiety complete the strychnan skeleton from which, via the Wieland-Gumlich aldehyde, the synthesis of (-)-strychnine is achieved. PMID- 10807176 TI - Membrane properties of archaeal macrocyclic diether phospholipids. AB - Several biophysical properties of four synthetic archaeal phospholipids [one polyprenyl macrocyclic lipid A and three polyprenyl double-chain lipids (B, C, D) bearing zero, one or four double bonds in each chain] were studied using differential scanning calorimetry, electron and optical microscopies, stopped flow/light scattering and solid-state 2H-NMR techniques. These phospholipids gave a variety of self-organized structures in water, in particular vesicles and tubules. These assemblies change in response to simple thermal convection. Some specific membrane properties of these archaeal phospholipids were observed: They are in a liquid-crystalline state over a wide temperature range; the dynamics of their polyprenyl chains is higher than that of n-acyl chains; the water permeability of the membranes is lower than that of n-acyl phospholipid membranes. It was also found that macrocyclization remarkably improves the barrier properties to water and the membrane stability. This may be related to the adaptation of Methanococcus jannaschii to the extreme conditions of the deep sea hydrothermal vents. PMID- 10807179 TI - Chiral allyl cations are captured by furan with 100% stereoselectivity: synthesis of enantiopure 2-alkoxy-8-oxabicyclo AB - A low-temperature (-95 degrees C) protocol for intermolecular cycloadditions of furan to chiral silyloxyallyl cations in dichloromethane is described. Key precursors are open-chain, mixed a-ketoacetals, which are chiral. The resulting [4+3] cycloadducts are densely functionalized and are isolated as single enantiomers in high chemical yield. The yield of the cycloadducts increases with increasing dilution. Three and four stereogenic centres are created in one single step. PMID- 10807178 TI - Synthesis and biological evaluation of integrin antagonists containing trans- and cis-2,5-disubstituted THF rings. AB - The synthesis of a series of RGD mimetics is described. All compounds consist of a central 2,5-disubstituted tetrahydrofuran core, a variable linker to a guanidino group, and a beta-amino alanine unit to mimic the carboxylic acid. Three types of linkers were investigated: a simple four-atom methylene chain (type A, compounds 14, 15, 16, and 17), a four-atom methylene chain with an additional chiral center, and a nitrogen substituent (type B, compounds 38, 39, and 40), and an amide linker of different length with an additional chiral center (type C, compounds 59, 60, 61, and 62). A variety of compounds were tested as potential integrin antagonists in a receptor binding assay (alphaIIbbeta3, alphavbeta3, and alphavbeta5). The relative and absolute configuration of the chiral centers at the THF ring had a pronounced effect on the binding activity and selectivity. Compound 14 proved to be a selective inhibitor of alphaIIbbeta3 (IC50=20nM), whereas compound 40 exhibited high activity for binding of alphaIIbbeta3 (IC50=67nM) and alphavbeta3 (IC50=52nM). PMID- 10807180 TI - Cooperative reactivity of early-late heterodinuclear transition metal complexes with polar organic substrates AB - A comprehensive investigation into the cooperative reactivity of two chemically complementary metal-complex fragments in early-late heterodinuclear complexes has been carried out. Reaction of the partially fluorinated tripodal amidozirconium complexes [HC-(SiMe2NR)3Zr(mu-Cl)2Li(OEt2)2] (R = 2-FC6H4: 2a, 2,3,4-F3C6H4: 2b) with K[CpM(CO)2] (M=Fe, Ru) afforded the stable metal-metal bonded heterodinuclear complexes [HC[SiMe2NR]3-Zr-MCp(CO)2] (3-6). Reaction of the dinuclear complexes with methyl isonitrile as well as the heteroallenes CO2, CS2, RNCO and RNCS led to insertion into the polar metal-metal bond. Two of these complexes, [HC[SiMe2N(2-FC6-H4)]3Zr(S2C)Fe(CO)2Cp] (9a) and [HC-[SiMe2N(2 FC2H4)]3Zr-(SCNPh)Fe(CO)2-Cp] (12), have been structurally characterized by a single crystal X-ray structure analysis, proving the structural situation of the inserted substrate as a bridging ligand between the early and late transition metal centre. The reactivity towards organic carbonyl derivatives proved to be varied. Reaction of the heterobimetallic complexes with benzyl and ethylbenzoate led to the cleavage of the ester generating the respective alkoxozirconium complexes [HC[SiMe2N(2-FC6H4)]3ZrOR] (R = Ph-CH2: 13a, Et: 13b) along with [CpFe [C(O)Ph](CO)2], whereas the analogous reaction with ethyl formate gave 13b along with [CpFeH(CO)2]; this latter complex results from the instability of the formyliron species initially formed. Aryl aldehydes were found to react with the Zr-M complexes according to a Cannizzaro disproportionation pattern yielding the aroyliron and ruthenium complexes along with the respective benzoxyzirconium species. The transfer of the aldehyde hydrogen atom in the course of the reaction was established in a deuteriation experiment. [HC[SiMe2-N(2-FC6H4)]3Zr-M(CO)2Cp] reacted with lactones to give the ring-opened species containing an alkoxozirconium and an acyliron or acylruthenium fragment; the latter binds to the early transition metal centre through the acyl oxygen atom, as evidenced from the unusuallly low-field shifted 13C NMR resonances of the RC(O)M units. Ketones containing a-CH units react with the Zr-Fe complexes cooperatively to yield the aldol coupling products coordinated to the zirconium complex fragment along with the hydridoiron compound [CpFeH(CO)2], whereas 1,2-diphenylcyclopropenone underwent an oxygen transfer from the keto group to a CO ligand to give a linking CO2 unit and a cyclopropenylidene ligand coordinated to the iron fragment in [HC [Si(CH3)2N(2,3,4-F3C6H2)]3Zr(mu-O2C)-Fe(CO)[C3Ph2)Cp] (19). The atom transfer was established by 17O and 13C labelling studies. Similar oxygen-transfer processes were observed in the reactions with pyridine N-oxide, dimethylsulfoxide and methylphenylsulfoxide. PMID- 10807181 TI - Carboxylate binding by 2-(guanidiniocarbonyl)pyrrole receptors in aqueous solvents: improving the binding properties of guanidinium cations through additional hydrogen bonds. AB - A series of guanidiniocarbonyl pyrrole receptors has been synthesized which bind carboxylates by ion pairing in combination with multiple hydrogen bonds. Their binding properties with various carboxylates have been investigated using NMR titration studies in 40% water/DMSO (v/v). The best receptor has association constants which are in the order of K approximately/= 10(3) mol(-1) and hence some 30 times larger than with the simple acetyl guanidinium cation. Through a systematic variation of the receptor structure, semiquantitative estimates for the energetic contributions of the individual binding interactions could be derived. These data show that the various hydrogen bonds are not equally important for the binding but differ significantly in their energetic contribution to the overall complexation process. Furthermore, the receptor can be made chiral and shows selectivity upon binding of enantiomeric amino acid carboxylates. Molecular modeling was used to obtain structural information for the various receptor carboxylate complexes and served as a basis to explain the observed differences in binding constants. PMID- 10807182 TI - The rates of the exchange reactions between [Gd(DTPA)]2- and the endogenous ions Cu2+ and Zn2+: a kinetic model for the prediction of the in vivo stability of [Gd(DTPA)]2-, used as a contrast agent in magnetic resonance imaging. AB - The kinetic stability of the complex [Gd(DTPA)]2- (H5DTPA = diethylenetriamine N,N,N',N",N"-pentaacetic acid), used as a contrast-enhancing agent in magnetic resonance imaging (MRI), is characterised by the rates of the exchange reactions that take place with the endogenous ions Cu2+ and Zn2+. The reactions predominantly occur through the direct attack of Cu2+ and Zn2+ on the complex (rate constants are 0.93+/-0.17 M(-1) s(-1) and (5.6+/-0.4) x 10(-2)M(-1) S(-1), respectively). The proton-assisted dissociation of [Gd(DTPA)]2- is relatively slow (k1 = 0.58+/-0.22 M(-1) s(-1)), and under physiological conditions the release of Gd3+ predominantly occurs through the reactions of the complex with the Cu2+ and Zn2+ ions. To interpret the rate data, the rate-controlling role of a dinuclear intermediate was assumed in which a glycinate fragment of DTPA is coordinated to Cu2+ or Zn2+. In the exchange reactions between [Gd-(DTPA)]2- and Eu3+, smaller amounts of Cu2+ and Zn2+ and their complexes with the amino acids glycine and cysteine have a catalytic effect. In a model of the fate of the complex in the body fluids, the excretion and the "dissociation" of [Gd(DTPA)]2- are regarded as parallel first-order processes, and by 10 h after the intravenous administration the ratio of the amounts of "dissociated" and excreted [Gd(DTPA)]2 is constant. From about this time, 1.71% of the injected dose of [Gd(DTPA)]2- is "dissociated". The results of equilibrium calculations indicate that the Gd3+ released from the complex is in the form of Gd3+-citrate. PMID- 10807183 TI - Oriented crystalline monolayers and bilayers of 2 x 2 silver(I) grid architectures at the air-solution interface: their assembly and crystal structure elucidation AB - Oriented crystalline monolayers, approximately 14 A thick, of a 2 x 2 Ag+ grid complex, self-assembled at the air-solution interface starting from an water insoluble ligand 3,6-bis[2-(6-phenylpyridine)]pyridazine spread on silver-ion containing solutions, were examined by grazing-incidence X-ray diffraction and specular X-ray reflectivity using synchrotron radiation. The monolayer structure was refined, including a determination of the positions of the counter-ions, with the SHELX-97 computer program. The monolayers were transferred from the interface onto various solid supports and visualized by scanning force microscopy, and characterized by X-ray photoelectron spectroscopy in terms of molecular structure. On surface compression, the initial self-assembled monolayer undergoes a transition to a crystalline bilayer in which the two layers, almost retaining the original arrangement, are in registry. Such a phase transition is of relevance to the understanding of crystal nucleation. PMID- 10807184 TI - Coincidence of the molecular organization of beta-substituted oligothiophenes in two-dimensional layers and three-dimensional crystals AB - The molecular arrangements of three different alkyl-substituted oligothiophenes both in two-dimensional adsorbed layers at a substrate interface and in bulk three-dimensional crystals were studied. Scanning tunneling microscopy (STM) was used to investigate the ordering of the conjugated oligomers in two-dimensional layers adsorbed on graphite. These data were compared with the X-ray structure determinations of single crystals revealing the arrangement in the three dimensional bulk material. Quaterthiophenes 1 and 2, bearing dodecyl and hexyl side chains, respectively, exhibit a lamella-type stacking of the conjugated backbone concomitant with an interlocking of the alkyl side chains both on the surface and in the crystal. In contrast, the arrangement of propyl-substituted quaterthiophene 3 is rather "herringbone-like" due to the reduced interactions of the shorter alkyl side chains. In all three cases, evidently, the two-dimensional ordering at the graphite surface is coincident with the molecular packing in one cross-section of the three-dimensional crystal. PMID- 10807185 TI - HIV-1 envelope protein gp41 modulates expression of interleukin-10 and chemokine receptors on monocytes, astrocytes and neurones. AB - OBJECTIVE: To analyse the effect of HIV-1 transmembrane protein gp41 on cytokine production and chemokine receptor expression in blood and brain. DESIGN: Because previous results had demonstrated that recombinant gp41 contributes to HIV induced dysfunction of blood immune cells we investigated its effect on interleukin (IL)-10 synthesis and expression of the HIV coreceptors CCR5 and CXCR4 in different human brain cells. METHODS: Astrocytic, microglial and neuronal cell lines were incubated with the extracellular domain of gp41 (aa565 647). Secretion of IL-10 into the medium was measured by ELISA. Chemokine receptor expression was analysed by fluorescence activated cell sorting and by RT PCR. RESULTS: Incubation of the astrocytic cell line U87 with gp41 induced more than a 10 fold up-regulation of IL-10 secretion. This modulation was shown to be time- and dose-dependent. Use of inhibitors for different signal transduction pathways indicated a similar transduction cascade for the alteration of IL-10 production in astrocytes as in monocytes with participation of cAMP/adenylate cyclase and activation of p70S6 kinase. To a lesser extent IL-10 synthesis was also up-regulated by gp41 in the neuronal cell line SK-N-SH. In all cell types up regulation of IL-10 paralleled by an enhanced expression of the chemokine receptor and HIV-1 coreceptor CCR5. This up-regulation was driven by IL-10 as shown by use of an IL-10 antibody. Expression of the chemokine receptor CXCR4 was only slightly altered. CONCLUSIONS: These findings suggest a role for gp41 in the modulation of brain-specific host defence, cell migration and cell infectivity by HIV. PMID- 10807186 TI - An early expansion of CD8alphabeta T cells, but depletion of resident CD8alphaalpha T cells, occurs in the intestinal epithelium during primary simian immunodeficiency virus infection. AB - OBJECTIVES: To evaluate changes in the phenotypic heterogeneity and function of CD8 T cells in the intestinal epithelium during primary SIV infection. DESIGN: Previous studies have shown an increased prevalence of CD8 T cells in the intestinal epithelium in HIV and SIV infections. As intestinal CD8 T cells are a heterogeneous population we evaluated their phenotypic distribution (CD8alphabeta, CD8alphaalpha) and function [interferon (IFN)-gamma production] during primary SIV infection. METHODS: The phenotype and functional potential of CD8 intestinal intraepithelial lymphocytes (IEL) prior to and following SIV infection were determined using flow cytometry. RESULTS: IEL were found to harbor CD8alphabetaCD3, CD8alphaalphaCD3 and CD8alphaalpha+CD3- T-cell subsets. Most of the CD8CD4 double positive IEL expressed CD8alphaalpha homodimers. In primary SIV infection the frequency of CD8alphabetaCD3 T cells increased dramatically whereas the frequency of CD8alphaalpha T cells declined. A higher frequency of CD8alphabetaKi-67 IEL was observed following SIV infection suggesting that local cell proliferation might have contributed to an increased prevalence of CD8alphabeta IEL. In contrast, a severe depletion of CD8alphaalphaCD4 IEL occurred which contributed to the depletion of CD8alphaalpha IEL. The CD8alphabeta IEL were the major producers of IFN-gamma in the intestinal epithelium and the frequency of IFN-gamma-producing CD8alphabeta IEL was enhanced considerably in primary infection. CONCLUSIONS: CD8alphabeta IEL may be important in generating early antiviral responses at the intestinal epithelium. However, alterations in CD8 T-cell subsets and their function may reflect early immunopathogenic events in the intestinal mucosa. PMID- 10807187 TI - HIV-infected human Langerhans cells transmit infection to human lymphoid tissue ex vivo. AB - OBJECTIVES: To create a novel ex vivo model for early biologic events involved in sexual transmission of HIV and to demonstrate that Langerhans cells (LC), the purported initial mucosal target cells for HIV, play a critical role in this process. METHODS: Epidermal cells containing LC were isolated from normal appearing skin of healthy volunteers and exposed to a panel of primary and laboratory-adapted R5- and X4-HIV isolates, washed and applied to the surfaces of allogeneic tonsil tissue blocks. Viral replication was followed by measuring HIV p24 protein in culture supernatants by ELISA. RESULTS: Both R5- and X4-HIV isolates could be transmitted by LC and established high levels of infection in lymphoid tissue (p24 > 10 ng/ml). Depletion of LC within epidermal cell suspensions abrogated the ability of HIV-exposed suspensions to transmit virus to tonsil histocultures. CONCLUSIONS: Using a novel ex vivo model, human LC are shown for the first time to be the major epidermal cell type that is involved in transmission of HIV infection to human lymphoid tissue. Importantly, this system could prove useful in further understanding LC trafficking and other early biological events involved in primary HIV infection. PMID- 10807188 TI - Specific recognition of lamivudine-resistant HIV-1 by cytotoxic T lymphocytes. AB - OBJECTIVE: The reverse transcriptase (RT) M184V mutation within the HLA-A2 restricted HIV-1 cytotoxic T lymphocyte (CTL) epitope VL9 (VIYQYMDDL; RT 179-187) not only induces drug escape against lamivudine but also abolished recognition by a CTL clone derived from a long-term non-progressor. To test whether the variant VL9 epitope containing the M184V mutation represents a new CTL epitope, we studied recognition of this epitope in a cohort of HLA-A2-positive HIV-1-infected patients. METHODS: Peripheral blood mononuclear cells isolated from 28 HIV-1 infected patients were stimulated with the peptide VIYQYVDDL, containing the M1 84V mutation. Outgrowing cell lines were tested for specific recognition in a standard chromium-release assay. RESULTS: In one subject, a CTL line could be isolated recognizing the peptide VIYQYVDDL in conjunction with HLA-A2. The CTL clone also recognized the M1841 mutation, but it failed to recognize the wild type epitope VIYQYMDDL. CONCLUSION: CTL can specifically recognize lamivudine resistant HIV-1 variants. Therefore, the cellular immune response could have an important influence on the control of drug-resistant virus. Furthermore, this demonstrates that the immune system can generate new CTL specificities even in patients with advanced disease, as the M184V HIV variants emerges only after drug treatment. Specific immunotherapy against this epitope might be helpful in delaying or preventing lamivudine resistance. PMID- 10807189 TI - Reduction of the HIV-1-infected T-cell reservoir by immune activation treatment is dose-dependent and restricted by the potency of antiretroviral drugs. AB - BACKGROUND: Treatments combining T-cell activating agents and potent antiretroviral drugs have been proposed as a possible means of reducing the reservoir of long-lived HIV-1 infected quiescent CD4 T-cells. OBJECTIVE: To analyse the effect of such therapies on HIV-1 dynamics and T-cell homeostasis. DESIGN AND METHODS: A mathematical framework describing HIV-1 dynamics and T-cell homeostasis was developed. Three patients who were kept on a particularly potent course of highly active antiretroviral therapy (HAART) were treated with the anti CD3 monoclonal antibody OKT3 and interleukin (IL)-2. Plasma HIV-RNA, and HIV-RNA and DNA in peripheral blood mononuclear cells and lymph node mononuclear cells were measured. These results and other published studies on the use of IL-2 alone were assessed using our mathematical framework. RESULTS: We show that outcome of treatment is determined by the relative rates of depletion of the infected quiescent T-cell population by activation and of its replenishment through new infection. Which of these two processes dominates is critically dependent on both the potency of HAART and also the degree of T-cell activation induced. We demonstrate that high-level T-cell stimulation is likely to produce negative outcomes, both by failing to reduce viral reservoirs and by depleting the CD4 T cell pool and disrupting CD4/CD8 T-cell homeostasis. In contrast, repeated low level stimulation may both aid CD4 T-cell pool expansion and achieve a substantial reduction in the long-lived HIV-1 reservoir. CONCLUSIONS: Our analysis suggests that although treatment that activates T-cells can reduce the long-lived HIV-1 reservoir, caution should be used as high-level stimulation may result in a negative outcome. PMID- 10807191 TI - Energy balance, viral burden, insulin-like growth factor-1, interleukin-6 and growth impairment in children infected with human immunodeficiency virus. AB - OBJECTIVE: To determine the relationship between energy metabolism and growth abnormalities in HIV-infected children and to assess clinical or laboratory characteristics which may be contributing factors to their growth impairment. DESIGN: A comparative study. METHODS: We measured energy intake by inpatient calorie count/outpatient 24 h food recalls, resting energy expenditure by indirect calorimetry, total energy expenditure by the doubly-labeled water technique, iron metabolism, protein metabolism, and lipid metabolism markers as well as CD4 count, viral load, insulin-like growth factor-1 (IGF-1), serum interleukin-6 (IL-6), and whole blood stimulated IL-6 levels in prepubertal congenitally HIV-infected children with normal and impaired growth patterns. RESULTS AND CONCLUSIONS: Differences in energy expenditures were not found between normal and growth-impaired HIV-infected children. Energy intake but not energy expenditure was significantly reduced when HIV-infected children were compared to expected normal values for age and gender. Advanced HIV clinical disease, severe immune suppression, increased viral burden, increased IL-6 activity, decreased total serum protein, and decreased IGF-1 levels were more likely to be found in HIV-infected children with growth impairment in comparison with HIV-infected children with normal growth. PMID- 10807190 TI - Efficacy, safety, and adherence with a twice-daily combination lamivudine/zidovudine tablet formulation, plus a protease inhibitor, in HIV infection. AB - OBJECTIVE: A randomized, open-label, multicenter study to establish clinical equivalence (non-inferiority) of a regimen employing a lamivudine 150 mg/zidovudine 300 mg combination tablet, administered twice daily, plus a marketed protease inhibitor, compared with a conventional regimen of 150 mg lamivudine twice daily, 600 mg zidovudine daily, and a protease inhibitor, in antiretroviral-experienced patients infected with HIV-1. PATIENTS: Adults who were seropositive for HIV-1 infection with plasma HIV-1 RNA levels < 10000 copies/ml (Roche Amplicor polymerase chain reaction assay, lower limit of quantitation (LLOQ) 400 copies/ml) and CD4+ cell counts > or = 300 x 10(6)/l). All patients had been receiving the conventional lamivudine/zidovudine/protease inhibitor regimen for > or = 10 weeks immediately prior to the study. INTERVENTION: Patients were randomized to the conventional regimen (n = 113) or combination tablet regimen (n = 110) for 16 weeks. The primary study endpoint was treatment failure, defined as an increase in HIV-1 RNA > or = 0.5 log10 above baseline in patients with viral load > LLOQ at randomization and as HIV-1 RNA increasing to > or = 1250 copies/ml in patients with viral load < LLOQ at randomization. RESULTS: The combination tablet regimen was associated with a 3.5% greater success rate than the conventional regimen (96.4 versus 92.9%), with four and eight patients failing treatment due to increases in HIV-1 RNA levels, respectively. The lower limit of the associated confidence interval for the difference was -2.4%, which was well within the -10% margin predefined as clinically unimportant. This establishes the clinical equivalence (non inferiority) of the combination tablet regimen to the conventional regimens regarding virologic response. The combination tablet and conventional regimens were similar with respect to percentage of patients maintaining HIV-1 RNA levels < LLOQ at the end of study or improving from baseline to undetectability (94 versus 91%; P= 0.063), overall incidence of drug-related adverse events (21 versus 19%) (P=0.868), and mean area under the curve for CD4+ cell counts [treatment difference, 5.9 cells (95% confidence interval, -15.8 to 27.6 x 10(6) cells/l)]. A self-reported adherence questionnaire indicated that patients in the combination tablet group were less likely to miss doses of nucleoside analogue medication at weeks 8 (P= 0.007) and 16 (P= 0.046). CONCLUSIONS: The combination lamivudine/zidovudine tablet/protease inhibitor regimen is clinically equivalent (non-inferior) to the conventional regimen with respect to virologic response and may offer adherence advantages. PMID- 10807193 TI - Assessing sexual risk behaviour of young gay men in primary relationships: the incorporation of negotiated safety and negotiated safety compliance. AB - OBJECTIVES: To establish the actual rate of risky unprotected anal intercourse (UAI) among young gay men in steady relationships by correcting for negotiated safety (NS) and NS compliance. In addition, to examine whether the rate of UAI with steady partners, after correction for NS and NS compliance, is higher than the rate of UAI during sexual contacts with casual partners. METHODS: A total of 435 young gay men completed questionnaires regarding relationship status, HIV status, NS agreements, and sexual behaviour with steady and casual partners in the preceding 6 months. RESULTS: Twelve per cent of the participants in a relationship practised NS. The non-compliance rate with NS agreements reached 10%. The rate of UAI with steady partners stood at 54%. After correction for NS and NS compliance, the actual risky UAI rate with steady partners dropped to 39%. Chi square tests showed that even after correction for NS and NS compliance, the rates of UAI with steady partners remained significantly higher than the rates of UAI with casual partners. CONCLUSION: It is imperative to correct the rates of UAI with steady partners for NS and NS compliance in order to estimate accurately the rates of risky UAI. The higher rates of risky UAI found with steady partners, even after correcting for NS, support the assumption that steady relationships provide a context that facilitates sexual risk-taking behaviour. We should therefore specifically target primary relationships as a source of risk for HIV transmission, and take into consideration non-compliance with NS agreements. PMID- 10807192 TI - Cost-effectiveness of cesarean section delivery to prevent mother-to-child transmission of HIV-1. AB - OBJECTIVE: To evaluate costs and outcomes of cesarean section performed before onset of labor and before rupture of membranes (elective cesarean section) compared to vaginal delivery among HIV-infected women. DESIGN: Cost-effectiveness and cost-benefit analysis. PARTICIPANTS AND SETTING: Pregnant HIV-infected women in the US who refrain from breastfeeding. INTERVENTION: Elective cesarean section versus vaginal delivery by antiretroviral therapy regimen. MAIN OUTCOME MEASURES: Pediatric HIV cases avoided, years of life saved, and direct medical costs for maternal interventions and pediatric HIV treatment. RESULTS: Elective cesarean section (versus vaginal delivery) was cost-effective among women receiving zidovudine prophylaxis (US$1131 per case avoided, US$17 per year of life saved) and combination antiretroviral therapy (US$112693 per case avoided, US$1697 per year of life saved), and cost saving among women receiving no antiretroviral therapy during pregnancy (benefit-cost ratio of 2.23). Although elective cesarean section remained cost-effective, results were sensitive to variations in vertical transmission rates and to pediatric HIV treatment costs. Population-based analyses indicated that elective cesarean section could prevent 239 pediatric HIV cases annually with a savings of over US$4 million. CONCLUSIONS: Elective cesarean section is a cost-effective intervention to prevent vertical transmission of HIV among women receiving various antiretroviral therapy regimens, who refrain from breastfeeding. PMID- 10807194 TI - Evidence that anoreceptive intercourse with ejaculate exposure is associated with rapid CD4 cell loss. AB - OBJECTIVE: To determine whether ejaculate exposure through anoreceptive intercourse is associated with rapid CD4 cell loss. DESIGN: Self-reported behavioral, demographic data and blood samples were gathered longitudinally at ten semiannual visits from individuals participating in the Multicenter AIDS Cohort Study (MACS). PATIENTS/PARTICIPANTS: A group of 937 HIV-seropositive men who were continuously followed for four to ten semiannual visits. OUTCOME MEASURES: A loss of 10% or more in CD4 cells between the first two of any three consecutive semiannual visits that was followed by a 10% or greater loss between the second and third visits. RESULTS: A period of rapid CD4 cell loss over three semiannual visits occurred in 389 of the 937 (42%) HIV-seropositive men studied. Men who reported one or more anoreceptive intercourse partners with whom they were exposed to ejaculate (RAI-E) during the 12 months immediately preceding their visits were more than twice as likely to show this rapid CD4 cell loss compared with men with no such partners. CONCLUSIONS: The association between RAI E partnerships and rapid CD4 cell loss suggests factors associated with ejaculate exposure (e.g., sexually transmitted diseases) may hasten the clinical progression of HIV disease. It is suggested that infectious diseases, which are known to be associated with ejaculate exposure, may be the causal factor underlying the association between RAI-E partnerships and rapid CD4 cell loss in these men, although the presence of these diseases was not ascertained in these data. HIV-infected individuals should be cautioned against unprotected anoreceptive intercourse. PMID- 10807195 TI - Correlates of HIV infection among young adult short-term injection drug users. AB - OBJECTIVES: To identify risks associated with HIV infection among young adult short-term injection drug users. METHODS: Current injection drug users, between 18 and 29 years of age, were recruited through street outreach to participate in a cross-sectional survey of HIV prevalence by circumstances of drug injection initiation, HIV-related risk behaviors, and a follow-up to estimate HIV incidence. RESULTS: At enrollment, 33 (14.4%) of 229 participants were HIV seropositive. Significant bivariate associations with HIV at the time injection drug use was initiated included age less than or equal to 18 years, having receptive anal sex with the person who assisted with initiation, and having two or more 'trainers' before being able to self-inject. Injecting risks positively associated with HIV included cocaine or speedball (heroin and cocaine together) injection versus heroin or amphetamine injection, injecting five or more times per day, daily crack smoking, backloading, sharing needles at peak drug use, and not using a new needle for every injection. Sexual practices associated with HIV included reporting > 100 lifetime sex partners, a history of sexual assault, being gay or bisexual, and trading sex for money or drugs after starting to inject. In a multivariate model, trading anal sex for money or drugs after initiating injection drug use [odds ratio (OR), 14.2; 95% confidence interval (CI) 3.2-62.3], cocaine/speedball injection (OR, 10.3; 95% CI, 2.2-47.9), daily crack smoking (OR, 4.2; 95% CI, 1.7-10.5), and having two or more trainers (OR, 2.6; 95% CI, 1.1 - 5.9) were independently associated with HIV. During 12 months of follow-up, four persons seroconverted for HIV (annual incidence: 2.6%; 95% CI, 1.1 -5.9%) CONCLUSIONS: Among short-term injectors, both sexual and injecting practices were important predictors of HIV infection, indicating that a proportion of HIV infections among young injection drug users can be attributed to sexual transmission. The incidence rate for HIV infection suggests that immediate steps should be taken to prevent new infections among young injection drug users. PMID- 10807196 TI - Prevalence of genotypic resistance to nucleoside analogues and protease inhibitors in Spain. The ERASE-2 Study Group. AB - OBJECTIVE: To examine the prevalence of resistance mutations to nucleoside reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) in a representative HIV-1 population in Spain. METHODS: A cross-sectional study was conducted including 601 HIV-infected patients who attended 20 Spanish hospitals in June 1998. Drug resistant mutations were examined using hybridization line probe assays (LiPA). The 6 bp insert at position 69 and the codon 75 mutant were examined by sequencing analysis in specimens lacking reactivity to 69/70 and 74 bands on LiPA, respectively. RESULTS: Primary resistance to NRTI was recognized in nine out of 52 (17%) naive individuals, whereas primary resistance to PI was found in seven out of 126 (6%) PI-naive patients. The codons most frequently involved in NRTI resistance were at positions 70 (66%), 184 (44%), 215 (33%), and 41 (11%), whereas the most common PI resistance mutation was at codon 82 (6/7 subjects). In pre-treated patients, the overall prevalence of resistant genotypes was 72.9% for NRTI and 27.2% for PI. The most frequent NRTI mutations occurred at codons 184 (38.5%), 215 (30.1%), and 41 (22.5%), whereas the most frequent PI mutations in pre-treated subjects were found at positions 82 (15.8%) and 84 (11.4%). Overall, patients who began triple combinations as initial therapy showed a lower number of key resistance mutations than those who began highly active antiretroviral therapy (HAART) after being exposed to NRTI for a period of time (mean number of mutations, 0.1 versus 1.8, P< 0.05). Codon 75 mutant was found in three out of 387 patients (0.7%), whereas no insertions at codon 69 were recognized. CONCLUSION: The prevalence of primary genotypic resistance to NRTI and PI in Spain was 17% and 6%, respectively. Zidovudine, lamivudine, indinavir and ritonavir were the drugs most frequently affected. These data support the use of resistance testing prior to the introduction of first-line antiretroviral therapies in Spain. Among pre-treated subjects, drug resistance genotypes were less prevalent in those who began HAART as initial therapy. PMID- 10807197 TI - Condom acceptance is higher among travelers in Uganda. AB - OBJECTIVE: To examine the role played by mobile residents in the spread of HIV through rural Uganda. DESIGN: Travel history and sexual network data were collected from a random sample of 1627 residents aged 15-49 years in Rakai District, Uganda during 1994. METHODS: Travelers and non-travelers are compared with respect to socio-demographic attributes, risk exposure, knowledge, attitudes and use of condoms using descriptive statistics and multivariate logistic regression. A demographic profile of travelers' partners is developed using information from a local network survey module. RESULTS: The population is highly mobile, with over 70% reporting travel to a potentially higher risk destination in the past year. Travelers are somewhat more likely to have higher levels of sexual risk behavior, but the risk appears to be offset by significantly greater knowledge, acceptance, and use of condoms. In multivariate analysis, the sexual risk differential for travelers is explained by occupational exposure and higher socio-economic status. The differential in condom acceptance, by contrast, appears to be associated with travel itself. Condom use with non-spousal partners is three times higher among travelers than non-travelers (P< 0.001), and travel remains a significant predictor after controlling for age, education, residence, occupation and multiple partners. Travelers are more likely to use condoms with both their local and non-local partners. Partners of male travelers are likely to be younger and better educated than those of male non-travelers. CONCLUSIONS: The mobile population in this rural region appears willing to adopt risk reduction measures appropriate to their exposure. This suggests that targeting condom promotion programs to travelers and their partners is likely to be effective in reducing the spatial diffusion of HIV, and may be an efficient method for spreading behavioral change into rural areas. PMID- 10807198 TI - Highly active antiretroviral therapy inhibits cytokine production in HIV uninfected subjects. PMID- 10807199 TI - Impaired induction of IL-15 in response to herpes simplex virus type 1 infection in peripheral blood mononuclear cells of HIV-infected patients. PMID- 10807200 TI - Absence of virological changes after acute febrile illnesses in two patients with undetectable pre-illness plasma HIV-1-RNA levels. PMID- 10807201 TI - Stromal derived factor-1alpha induces apoptosis in activated primary CD4+ T cells. PMID- 10807202 TI - Molecular screening for HIV-1 group N and simian immunodeficiency virus cpz-like virus infections in Cameroon. PMID- 10807203 TI - Transient rebound of plasma HIV-1 RNA is not followed by repopulation of the lymphoid compartment with HIV-1-infected cells. PMID- 10807204 TI - Distinguishing efficacy, individual effectiveness and population effectiveness of therapies. PMID- 10807205 TI - HIV prevention information, motivation, behavioral skills and behaviour among truck drivers in Chennai, India. PMID- 10807206 TI - Highly active antiretroviral therapy does not improve survival of patients with high JC virus load in the cerebrospinal fluid at progressive multifocal leukoencephalopathy diagnosis. PMID- 10807207 TI - Types of marijuana users by longitudinal course. AB - OBJECTIVE: Taxonomies of alcoholism and antisocial behaviors based on developmental course converge on two-group classifications that emphasize early and late onset. Typologies for users of illicit drugs remain to be developed. This article proposes a developmental taxonomy of marijuana users. METHOD: Cluster analysis was applied to a representative community sample of 708 (364 male, 344 female) marijuana users followed from adolescence to age 34-35. The Ward method, followed by relocation, was used to classify marijuana users into different types based on age of onset, chronicity of heavy use and persistence of use. ANOVA and logit analyses were utilized to describe the cluster solution and examine the correlates of cluster membership. RESULTS: Four marijuana use clusters were identified: early onset-heavy use, early onset-light use, mid onset heavy use and late onset-light use. The groups differed from each other in degree of involvement in marijuana and other drugs, sociodemographic and lifestyle characteristics. The majority of those with early onset did not become heavily involved in marijuana. Unique factors were associated with membership in each group. Factors differentiating early from mid-onset heavy use included association with marijuana-using peers and having had a mental disorder. Peer delinquency was an additional factor differentiating early initiators who became heavy users from those who did not. CONCLUSIONS: A simple two-type classification fails to take into account the heterogeneity of early and late onset groups. By itself, early onset into marijuana will not lead to problematic use or rapid progression into the use of other drugs. Motivation underlying use and dysfunctional behaviors are associated with the development of problematic drug use and dependence. PMID- 10807208 TI - Role of general and specific competence skills in protecting inner-city adolescents from alcohol use. AB - OBJECTIVE: The purpose of this longitudinal investigation was to test whether higher levels of general competence are linked to greater refusal assertiveness that is, in turn, related to less subsequent alcohol use among inner-city adolescents. METHOD: A large sample of students attending 22 middle and junior high schools in New York City participated. Students completed surveys at baseline, at 1-year follow-up and at 2-year follow-up (N = 1,459; 54% female). The students self-reported alcohol use. decision-making skills, self-efficacy and refusal assertiveness. Teams of three to five data collectors administered the questionnaire following a standardized protocol. The data were collected in school during a regular 40-minute class period. RESULTS: According to the tested structural equation model, Decision Making (beta = .07, p < .05) and Self Efficacy (beta = .24, p < .001) predicted higher Refusal Assertiveness and this greater assertiveness predicted less drinking at the 2-year follow-up (beta = .21, p < .001). Earlier drinking predicted 2-year follow-up drinking (beta = .40, p < .001). Goodness-of-fit indices were excellent (chi2 = 1107.9, 238 df, N = 1,438, p < .001; NFI = .93, NNFI = .94, CFI = .95). CONCLUSIONS: The tested model had a good fit and was parsimonious and consistent with theory. This research highlights the importance of addressing decision-making skills, self-efficacy and refusal assertiveness within adolescent alcohol prevention programs. PMID- 10807209 TI - Alcohol-related relative risk of driver fatalities and driver involvement in fatal crashes in relation to driver age and gender: an update using 1996 data. AB - OBJECTIVE: To re-examine and refine estimates for alcohol-related relative risk of driver involvement in fatal crashes by age and gender as a function of blood alcohol concentration (BAC) using recent data. METHOD: Logistic regression was used to estimate age/gender specific relative risk of fatal crash involvement as a function of the BAC for drivers involved in a fatal crash and for drivers fatally injured in a crash, by combining crash data from the Fatality Analysis Reporting System with exposure data from the 1996 National Roadside Survey of Drivers. RESULTS: In general, the relative risk of involvement in a fatal vehicle crash increased steadily with increasing driver BAC in every age/gender group among both fatally injured and surviving drivers. Among 16-20 year old male drivers, a BAC increase of 0.02% was estimated to more than double the relative risk of fatal single-vehicle crash injury. At the midpoint of the 0.08% - 0.10% BAC range, the relative risk of a fatal single-vehicle crash injury varied between 11.4 (drivers 35 and older) and 51.9 (male drivers, 16-20). With only very few exceptions, older drivers had lower risk of being fatally injured in a single-vehicle crash than younger drivers, as did women compared with men in the same age range. When comparable, results largely confirmed existing prior estimates. CONCLUSIONS: This is the first study that systematically estimated relative risk for drink-drivers with BACs between 0.08% and 0.10% (these relative risk estimates apply to BAC range midpoints at 0.09%.) The results clearly show that drivers with a BAC under 0.10% pose highly elevated risk both to themselves and to other road users. 2000) PMID- 10807210 TI - Gender differences in the control of alcohol-impaired driving in California. AB - OBJECTIVE: An examination of gender differences in alcohol-impaired driving. METHOD: Attitudes, perceptions and behaviors concerning drinking and driving were compared for male and female California drivers in three random-digit-dialing telephone survey interviews from 1983, 1986 and 1994 (survey response rates of 58%, 52% and 49%, respectively). A moral reasoning framework was applied to account for observed gender differences. RESULTS: Gender differences were examined within aggregate data from the 1983 and 1986 surveys (n = 291), and within the 1994 survey data (n = 608). Self-reported drinking-driving violations showed a substantial decline for both men and women across the survey periods (although violations remained much higher for men), paralleling the well documented drop in alcohol-related traffic crashes during this time span. Men and women responded equally to the threat of punishment from the legal system (threat of arrest, jail, loss of license, fine, increased insurance), but women were much more responsive to social and internal controls (perceived disapproval from friends, feelings of guilt, violation of a moral standard). CONCLUSIONS: These gender differences suggest that women may play an important role in strengthening broad social and moral constraints regarding alcohol-impaired driving. Implications of these findings to deterrence, the effects of sanctions, and social change are discussed. PMID- 10807211 TI - Reducing harmful alcohol-related behaviors: effective regulatory methods. AB - OBJECTIVE: This article examines the effects of tort liability, criminal law, administrative regulation, price and availability of alcohol, and personal and state characteristics on the decisions to engage in heavy episodic drinking and to drink and drive. METHOD: Individual behavior data from the Behavioral Risk Factor Surveys (1984-95) were used in a logit analysis of the probability that a respondent engaged in heavy episodic drinking (n = 86,273), drinking and driving (n = 87,087) and drinking and driving if also a heavy episodic drinker (n = 22,261). RESULTS: Imposing tort liability on bars reduced self-reported incidents of drunk driving among all drinkers (p = .043) but did not reduce the probability of heavy episodic drinking or drinking and driving among heavy drinkers. In this first national study of the impact of social host liability, we found that such liability lowered the self-reported probability of heavy episodic drinking (p = .0004) and drinking and driving among all drinkers (p = .0005). CONCLUSIONS: Although several criminal and administrative regulations were also effective in reducing heavy episodic drinking and drunk driving, the imposition of tort liability represents a useful addition to the arsenal of alcohol-control policies. PMID- 10807212 TI - Alcohol use, condom use and partner type among heterosexual adolescents and young adults. AB - OBJECTIVE: The present study examined the links between alcohol use and condom use, and the nature of the sexual relationship in which these behaviors occurred, and tested the notion that partner type suppresses the negative, direct effects of alcohol use on condom use. METHOD: Data were collected as part of face-to-face interviews conducted with 1,417 (54% female) randomly selected young adults (aged 18-25 years) who had had sex in the past 6 months. RESULTS: Using within-subjects analytic procedures, results showed that both drinking and condom use were more common with casual than with serious sexual partners, as expected. Moreover, consistent with the suppression hypothesis, the relationship between alcohol use and condom use became significant and negative only after controlling for partner type. CONCLUSIONS: These findings suggest that the interrelationships among partner type, alcohol use and condom use are complex and that the processes linking each pair of variables are best understood as part of a larger system of interconnected variables. PMID- 10807213 TI - Drinking and DSM-IV alcohol and drug dependence among white and Mexican-American DUI offenders. AB - OBJECTIVE: To report data on drinking patterns and DSM-IV alcohol and drug dependence among whites and Mexican Americans in treatment for driving under the influence of alcohol (DUI). METHOD: Interviews were conducted with a sample of 250 whites and 250 Mexican Americans admitted to one of five DUI treatment programs in a Northern California county. Interviews averaged 1 hour in length and were conducted in the programs by trained interviewers. The response rate was 72% for whites and 83% for Mexican Americans. RESULTS: Whites drink most frequently, followed by U.S.-born Mexican Americans and, finally, by Mexican Americans born in Mexico. However, rates of the amount of alcohol usually consumed are higher for Mexican Americans born in Mexico than for the other two groups. Prevalence rates of DSM-IV alcohol dependence are: 29% for whites, 27% for U.S.-born Mexican Americans and 9% for those born in Mexico. Prevalence rates of DSM-IV drug dependence are: 23% for whites, 24% for U.S.-born Mexican Americans and 6% for those born in Mexico. CONCLUSIONS: Rates of alcohol and drug dependence in these DUI treatment programs are much higher than rates of dependence in the general population, independent of ethnicity. Patterns of alcohol consumption vary dramatically among whites, U.S.-born Mexican Americans and Mexican Americans born in Mexico. The large amounts of alcohol ingested per occasion by Mexican-born Mexican Americans is particularly noteworthy and may put them at more risk for DUI. However, drug use and drug dependence are higher among whites and U.S.-born Mexican Americans than among Mexican Americans born in Mexico. PMID- 10807214 TI - Alcohol abuse and dependence in a national sample of psychiatric patients. AB - OBJECTIVE: To examine clinical characteristics and services being provided to Alcohol Abuse/Dependent (AAD) patients in current psychiatric practice. METHOD: In a national sample of psychiatrists (N = 417), each provided data on three preselected patients (N = 1,245; 51.8% women) that included demographics, DSM-IV diagnoses, treatment setting and health-plan measures. Logistic regression was used to compare patients with and without an AAD diagnosis. RESULTS: Only 12% of patients (n = 151) had an AAD diagnosis. AAD patient care was more frequently subject to utilization review and restriction or specification of medications to be prescribed (formulary). Psychiatrists also perceived greater restrictions on AAD patient care (e.g., requirements to use specific practice guidelines or treatment algorithms). CONCLUSIONS: Findings suggest that health care systems are subjecting treatment patients with AAD to greater scrutiny and may be limiting the extent and nature of care provided to these patients. The low prevalence of AAD among patients being seen by psychiatrists also warrants further attention. Study findings highlight the utility of practice-based research in addiction psychiatry. PMID- 10807215 TI - Withdrawal and tolerance: prognostic significance in DSM-IV alcohol dependence. AB - OBJECTIVE: The clinical and research importance of tolerance and/or withdrawal in the diagnosis of substance dependence has been identified as a key area needing clarification. Earlier longitudinal studies did not identify whether diagnoses of alcohol dependence were current or lifetime. In this study, the prognostic significance of the DSM-IV physiological specifier was investigated among cases of alcohol dependence current at the baseline interview. The role of tremors, required in DSM-III and DSM-III-R, was investigated as well. METHOD: Household residents (N = 130; 57% men) meeting full criteria for current DSM-IV alcohol dependence in a baseline interview were re-interviewed a year later and DSM-IV alcohol dependence was again evaluated. Reliable structured interviews were used to make the diagnoses. The DSM-IV physiologic specifier and its components were tested as predictors of outcome of DSM-IV alcohol dependence, and included an additional definition of alcohol withdrawal that required tremors. Chi-square tests and adjusted odds ratios indicated statistical significance of group differences on outcome. RESULTS: The DSM-IV physiological specifier (tolerance or withdrawal) did not predict 1-year chronicity of DSM-IV alcohol dependence, nor did tolerance when considered separately. However, withdrawal predicted poor outcome (e.g., 1-year chronicity of DSM-IV alcohol dependence). The effect of withdrawal was strengthened when tremors were required as part of the withdrawal syndrome. CONCLUSIONS: In contrast to earlier longitudinal research, this study supports the prognostic relevance of one component of the present DSM-IV physiological specifier--withdrawal. Withdrawal appears to have stronger prognostic meaning when the DSM-III-R version, in which tremors were required, is used. Further studies of the physiological subtype may benefit from studying subjects who have been clearly identified at the baseline evaluation as having a current diagnosis of dependence. PMID- 10807217 TI - A brief screening instrument for problem drinking in the emergency room: the RAPS4. Rapid Alcohol Problems Screen. AB - OBJECTIVE: While a number of brief screening instruments for identifying problem drinkers have been tested in clinical settings, instruments have not been found to perform as well for women as for men, or to perform uniformly across ethnic groups. The purpose of this study was to evaluate a shortened version of the RAPS (Rapid Alcohol Problems Screen) in an emergency room (ER) sample (N = 1,429; 51% female) and to determine the most efficient ordering of the items. METHOD: The sensitivity and specificity of each of the RAPS items were examined against current ICD-10 and DSM-IV criteria for alcohol dependence, and separately for harmful drinking or abuse. A four-item version of the RAPS (the RAPS4) was analyzed separately for men and for women, and for blacks, Hispanics and whites/others. RESULTS: Among the five original RAPS items, four items were found to be most efficient, with the single item of feeling guilt or remorse after drinking identifying 83% of those with alcohol dependence and 44% of those meeting criteria for harmful drinking or abuse. A positive response to any one of the four items (RAPS4) gave a sensitivity of 93% and specificity of 87% for alcohol dependence, and sensitivity and specificity were consistently high across gender and ethnic subgroups. Sensitivity and specificity for harmful drinking or abuse were lower (55% and 79%, respectively). CONCLUSIONS: Because of its brevity and high performance across demographic subgroups, the RAPS4 may hold promise in screening for alcohol use disorders in patient populations, and its utility warrants further evaluation in clinical settings. PMID- 10807216 TI - Diagnostic concordance of DSM-III, DSM-III-R, DSM-IV and ICD-10 alcohol diagnoses in adolescents. AB - OBJECTIVE: Little is known about the validity of diagnostic criteria for alcohol use disorders (AUDs) when applied to adolescents. This study examined the diagnostic concordance of DSM-III, DSM-III-R, DSM-IV and ICD-10 AUDs in a sample of adolescents with a broad range of alcohol problem severity. METHOD: Participants were 413 adolescents (250 male), ages 13 to 19, drawn from clinical and community sources. AUDs were assessed using the Structured Clinical Interview for the DSM (SCID), modified to make diagnoses in the four nosological systems. Diagnostic agreement for lifetime diagnoses was quantified with the kappa statistic. RESULTS: Agreement was fair to high across the three categories of alcohol dependence, alcohol abuse and no alcohol diagnosis (kappa = 0.51 to 0.76); for alcohol dependence (kappa = 0.51 to 0.83); and for the categories of any AUD versus no AUD (kappa = 0.55 to 0.96). Concordance was very low for alcohol abuse diagnoses (kappa = 0.10 to 0.23), with the exception of DSM-III-R and DSM-IV (kappa = 0.62). Dependence was superior to abuse in the degree of temporal overlap in diagnostic agreements. CONCLUSIONS: Similar to findings with adults, diagnostic concordance among adolescents tended to be fair to high for alcohol dependence and very low for alcohol abuse. The data highlight the inconsistency across nosological systems in the conceptual framework and definition of the alcohol abuse category. PMID- 10807218 TI - Self-reported frequency of drinking assessed with a closed- or open-ended question format: a split-sample study in Switzerland. AB - OBJECTIVE: This study was designed to determine whether reporting of drinking frequency differed according to whether the question asked was open or closed ended. METHOD: In a split-sample design with 994 participants responding to a questionnaire, the question on usual frequency of drinking for one sample (n = 457) was open ended, and for the other (n = 537) was closed ended (participants were offered seven response alternatives, ranging from "three times a day" to "never"). RESULTS: The closed-ended question elicited higher frequencies of drinking than the open-ended question. No significant interactions could be found between sociodemographic background variables and question format. CONCLUSION: This study does not support the general recommendation of survey methodologists that questions about frequency of behavior be open ended. PMID- 10807219 TI - What is a drinking episode? AB - OBJECTIVE: The phrase "drinking episode" is used informally in many ways. However, a scientific understanding of the factors affecting the length of drinking episodes and the way treatment components affect these episodes requires rigorous operational definitions, supported by evidence for the appropriateness of these definitions. METHOD: Daily drinking data from two studies (Project MATCH and BETA) involving a total of 1,955 subjects are examined by survival analysis methods to determine the prognostic significance of different durations of postdrinking abstinence. The dependent measures are "time to next drink" and "time to heavy drinking." RESULTS: Curves relating postdrinking abstinence to subsequent drinking indicate that 1 day of abstinence has little prognostic significance. As the duration of abstinence increases from 1 up to 60 days, longer abstinence has a decelerating but still positive association with time to subsequent drinking/heavy drinking. There is no apparent threshold point beyond which further abstinence has no further effect. Inflections in the curves suggest, however, that intervals of 1, 2 or 4 weeks of continuous abstinence may be important milestones. These general patterns seem to hold up across samples despite significant quantitative differences across studies. CONCLUSIONS: These results suggest that two different definitions of "drinking episode" may be useful in examining treatment effects on drinking behavior. These analyses help to provide a foundation for further quantitative research on treatment effects on addictive behaviors over time. PMID- 10807220 TI - Work stressors and alcohol consumption among white-collar workers: a signal detection approach. AB - OBJECTIVE: Previous findings regarding the relationship between work stress and alcohol consumption are inconsistent and inconclusive. One of the primary reasons for the inconsistency of the findings is that simple models, such as multiple regression analysis or analysis of variance (ANOVA), have been conventionally adopted in studies examining the relationship between work stressors and alcohol consumption. Since higher order interactions of multiple work stressors, which lead to heavy alcohol consumption, might not be fully uncovered within the framework of a multiple regression analysis or ANOVA, another approach to exploring the interactions of work stressors is presented in this study. METHOD: A signal detection analysis method was used to evaluate higher order interactions of work stress variables and to obtain basic information on 465 Japanese male white-collar workers. RESULTS: Of 11 work stressor or work position variables, four significant predictors of heavy alcohol consumption were identified. CONCLUSIONS: Two procedures were successfully performed in this study: (1) evaluating several higher order interactions of work stressors associated with heavy alcohol consumption, and (2) obtaining basic information to permit dividing the subjects into subgroups for further analysis using models and independent variables unique to each subgroup. PMID- 10807221 TI - Stress, alcohol use and alcohol-related problems: the influence of negative and positive affect in two cohorts of young adults. AB - OBJECTIVE: Past research has suggested that trait-based positive and negative affect mediate the relationships between self-report measures of stress and health. We extended these assumptions to the study of health-related behaviors, specifically the much studied stress-alcohol relationship. In addition, we sought to replicate previous findings showing that stress is more strongly associated with alcohol problems and not alcohol use as many theories (e.g., tension reduction hypothesis) have assumed. METHOD: Structural equation modeling was used to test a mediating variable model in two cohorts (574 and 239 subjects) of randomly selected young adults between 18 and 35 years of age. Participants were from the 1996 data wave of the Niagara Young Adult Health Study, a community based study incorporating a modified cohort-sequential design. The larger (older) cohort consisted of subjects initially contacted in 1990. RESULTS: For both cohorts, the measurement model proved to be very reliable. The final structural models did not match the predicted model. The correlations between the Stress and Alcohol Use latent variables (necessary to test a mediational model) were not significant in either cohort. However, in the older cohort, trait-based positive affect and hostility acted as intervening variables, allowing for an indirect relationship between stress and alcohol use. In the younger cohort, this indirect pattern was not present because none of the latent affect variables led to alcohol use. In both cohorts, stress predicted alcohol problems, even after controlling for the relationships between stress, affect and alcohol use. Significant cohort differences in the structural models were observed. CONCLUSIONS: The assumptions regarding the mediation of affect on the relationship between stress and health cannot be extended completely to the stress-alcohol relationship. Two reasons for this are: (1) the differences between the two age cohorts in the relationships between stress, affect and alcohol use and (2) the finding that stress is more predictive of alcohol-related problems than it is of alcohol use. PMID- 10807223 TI - Methodologies employed in the Journal of Studies on Alcohol. PMID- 10807222 TI - Typologies of the cultural position of drinking. AB - OBJECTIVE: Typologies of the cultural position of drinking from the social science literature are reviewed. METHOD: The article reviews significant studies and literature on the topic. RESULTS: Starting in the 1940s, two research traditions considered variations in the cultural position of drinking as explanations of rates of drinking problems. A "holocultural" tradition coded and analyzed ethnographic data on tribal and village societies, starting in the 1940s, with each study identifying a different social dimension as crucial. A "sociocultural" tradition distinguished abstinent cultures from prescriptive cultures, in which drinking was integrated with daily life, and expected, but drunkenness was prohibited. These types were implicitly contrasted with American drinking, which was variously characterized. Other dimensional and typological approaches in the literature are considered, including a little-known Jellinek typology. Problems with the widely used distinction between "wetter" and "drier" (or "temperance") cultures are discussed. CONCLUSIONS: Four ideal types of the cultural position of drinking can be readily distinguished: abstinent societies, constrained ritual drinking, banalized drinking and fiesta drunkenness. A large residual category remains, however, and a dimensional approach to typology building may be more fruitful. Two basic dimensions are proposed--regularity of drinking and extent of drunkenness--and further dimensions are described that may be added to fit the requirements of the particular study. PMID- 10807224 TI - Physiology of infants with very low birth weight. AB - Advances in neonatal management have resulted in dramatic increases in survival in infants with birth weights less than 1,500 g. Extensive basic science and clinical research has led to a more comprehensive understanding of the physiological differences between the VLBW infant and larger neonates. Meticulous attention must be paid to appropriate fluid, electrolyte, nutrition, and temperature maintenance to achieve homeostasis and growth. Additionally, the clinician must be aware of the diagnostic and treatment modalities for the common complications seen in the premature infant to minimize mortality and long-term morbidity. PMID- 10807225 TI - Ethical issues in treating infants with very low birth weight. AB - The purpose of this report is to provide pediatric surgeons with an ethical framework and a process for ethical decision making that can be applied to the difficult issues that arise in the care of infants with very low birth weight (VLBW). Clinical ethical issues focus around choices for surgical intervention, the use of total parenteral nutrition (TPN), recommendations for bowel transplantation, and management of dying infants. The role of family in decision making and the appropriate use of common distinctions including active or passive, withholding or withdrawing, and ordinary or extraordinary in decisions about life-sustaining treatments are discussed. A clinical case discussion illustrates the application of the process for ethical decision making. PMID- 10807226 TI - Necrotizing enterocolitis in infants with very low birth weight. AB - Necrotizing enterocolitis (NEC) is a disease in which the primary risk factor is prematurity. Despite, and partially as a result of, the tremendous strides neonatal care has taken, it is a major cause of morbidity and mortality of the newborn. The infant with very low birth weight is particularly susceptible, and the management of the condition in this group differs somewhat from other neonates. The outcomes continue to improve, but there are significant sequelae. Prevention, which would be the best "cure," is elusive, in no small part because of the multifactorial nature of the etiology of NEC. PMID- 10807227 TI - Esophageal atresia in infants with very low birth weight. AB - Infants with esophageal atresia (EA), with or without a tracheoesophageal fistula (TEF) frequently are of low birth weight. With advances in neonatal, respiratory, surgical, and anesthetic care, more infants with very low birth weight (VLBW; birth weight less than 1.5 kg) are surviving. The therapy of the VLBW neonate with EA is not longer automatically staged. Primary or delayed primary anastomosis can be performed safely if the patient is stable. This report will review the epidemiology, pathophysiology, treatment, and prognosis of EA in VLBW infants. The authors present their own experience in dealing with the VLBW with EA and review the world literature. PMID- 10807228 TI - Meconium diseases in infants with very low birth weight. AB - Retained meconium resulting in intestinal obstruction in infants with very low birth weight (VLBW) may cause significant morbidity and even mortality. Immature intestinal neuromuscular function, abnormal meconium composition, and maternal magnesium sulfate therapy results in delay of first meconium passage. This "normal" passage of first meconium may not occur until 1 week of life or later. A spectrum of conditions results from this delayed passage. At the most serious end of this continuum is intestinal obstruction. These patients present with abdominal distension, emesis, and reduced stool frequency. Early diagnosis of this condition using plain abdominal radiographs and contrast enemas where appropriate is imperative. Contrast enemas may be both diagnostic and therapeutic. Most patients do not need surgical intervention. Operation is indicated when enemas fail to relieve the obstruction or perforation occurs. Prompt diagnosis and appropriate management results in a good outcome. Meconium obstruction in VLBW infants does not appear to be associated with cystic fibrosis or Hirschsprung's disease. PMID- 10807229 TI - Inguinal hernia in infants with very low birth weight. AB - Inguinal hernias (IH) are among the most commonly encountered surgical problems in infants with very low birth weight (VLBW, <1,500 g) with a reported incidence of 16%. A trend toward earlier operation has emerged in recent years, with most now being repaired before discharge from the neonatal intensive care unit. The authors review the many special concerns regarding the management of IH in this patient population, including the timing of repair, the risk of incarceration, anesthetic management, the frequency of bilaterality, the high incidence of undescended testes, and the technical aspects and complications associated with IH repair in the VLBW infant. PMID- 10807230 TI - Abdominal wall defects in infants with very low birth weight. AB - Management strategies for infants with very low birth weight (VLBW) who have abdominal wall defects essentially are the same as for those in larger infants. The authors favor primary closure in infants with gastroschisis, and have achieved this goal in 91% of infants since 1985. Treatment of infants with omphalocele is based on the size of the defect and the presence of respiratory insufficiency or severe associated anomalies. Nonoperative treatment is used initially for infants with large defects or associated anomalies, with planned closure of the resultant ventral hernia when the infant weighs 20 pounds or is 1 year old. This technique helps avoid the complications associated with mechanical ventilation and with tight primary closure such as intestinal dysfunction and wound problems. PMID- 10807231 TI - Cardiac surgery in infants with very low birth weight. AB - Congenital heart defects in low birth weight infants are typically managed with supportive therapy or palliative surgery, and definitive repair is delayed. However, the morbidity of such an approach has been shown to be high. Recent reports emphasize early surgical repair with good results in low birth weight and premature babies. However, there are no data on the outcomes in infants with very low birth weight (<1,500 g). Since 1990, the authors performed complete repair of congenital heart defects (other than isolated patent ductus arteriosus) in 20 infants weighing 1,500 g or less. Defects included aortic coarctation (n = 6), ventricular septal defect (n = 5), tetralogy of Fallot (n = 2), transposition of great arteries (n = 2), truncus arteriosus (n = 2), total anomalous pulmonary venous return (n = 1), double outlet right ventricle (n = 1), and pulmonary atresia with intact ventricular septum (n = 1). Preoperative morbidity was more common in patients who were referred late for corrective surgery. Modification of techniques of neonatal cardiopulmonary bypass were used. There were 2 early deaths (10%) caused by foot gangrene (n = 1), and pulmonary complications (n = 1). No patient had evidence of postbypass intracranial hemorrhage. At a median follow-up of 40 months, there was 1 late death, and 4 patients underwent a total of 5 surgical and catheter reinterventions. There was no evidence of neurological sequelae attributable to surgery. In most cases, delaying repair of congenital heart defects in low and very low birth weight infants does not confer any benefit and is associated with a higher incidence of preoperative morbidity. Complete repair of both simple and complex congenital heart lesions can be performed successfully in such patients with good early and medium-term results. Postoperative growth after repair approximates the normal growth curve for low birth weight infants without congenital heart disease. It is recommended that with few exceptions, such infants, especially when symptomatic, undergo early surgical correction rather than prolonged medical management or other forms of palliation. PMID- 10807232 TI - Management of cholestasis in infants with very low birth weight. AB - Infants with very low birth weight (VLBW) are at increased risk of cholestasis when compared with older infants and children. Factors associated with this increased risk of cholestasis include immaturity of the biliary excretory system, a diminished immune response to sepsis, an increased incidence of necrotizing enterocolitis and short bowel syndrome, as well as an increased exposure to parenteral nutrition (PN). The current literature on cholestasis in VLBW infants and the factors that mediate the initiation and progression of cholestatic liver damage is reviewed. A protocol for managing infants with cholestatic jaundice is presented, and a case report is included that shows use of the protocol to normalize the bilirubin in a VLBW infant with severe cholestatic jaundice. PMID- 10807233 TI - Avoidance and treatment of retinopathy of prematurity. AB - Retinopathy of prematurity is a blinding eye disease of premature infants. It is increasing in incidence as more babies of lower birth weights survive. Although in the 1940s and 1950s the major predisposing factor was high oxygen exposure, the main risk factor now is birth weight less than 1,000 g. Retinopathy of prematurity can be classified into several distinct stages, which progress in a typical manner. Once the stage of threshold is reached, the eye has a 50% chance of becoming blind. At this stage, laser peripheral retinal photo ablation or peripheral cryo ablation may cause the neovascularization to regress and preserve vision. The treatment is not effective in all cases, however, and a large number of children continue to become blind each year because of this devastating disease. New microsurgical techniques such as lens-sparing vitrectomy and modified scleral buckle for tiny eyes have improved the outcome for some children. Prevention is the best hope to eradicate blindness caused by this disorder, and manipulations of the metabolic and neonatal intensive care unit environment are currently being studied. PMID- 10807234 TI - Brave new world: reaching for utopia. PMID- 10807235 TI - Molecular biology and neurosurgery in the third millennium. AB - The application of techniques in molecular biology to human neurosurgical conditions has led to an increased understanding of disease processes that affect the brain and to novel forms of therapy that favorably modify the natural history of many of these conditions. Molecular strategies are currently being either used or sought for brain tumors, stroke, neurodegenerative diseases, vascular malformations, spinal degenerative diseases, and congenital malformations of the central nervous system. Considering that the structure of deoxyribonucleic acid was ascertained by Watson and Crick as recently as 1953, the progress that has been made to implement molecular medicine in clinical practice has been meteoric. More than 2000 patients have been treated in approved gene therapy trials throughout the world. Many of these patients have been treated for neurological diseases for which conventional medical therapies have been of limited utility. As part of this continuing series on advances in neurosurgery in the third millennium, we first reflect on the history of the nascent field of molecular biology. We then describe the powerful techniques that have evolved from knowledge in this field and have been used in many publications in Neurosurgery, particularly within the past decade. These methods include commonly used techniques such as advanced cytogenetics, differential display, microarray technology, molecular cell imaging, yeast two-hybrid assays, gene therapy, and stem cell utilization. We conclude with a description of the rapidly growing field of bioinformatics. Because the Human Genome Project will be completed within 5 years, providing a virtual blueprint of the human race, the next frontier (and perhaps our greatest challenge) will involve the development of the field of "proteomics," in which protein structure and function are determined from the deoxyribonucleic acid blueprint. It is our conviction that neurosurgeons will continue to be at the forefront of the treatment of patients with neurological diseases using molecular strategies, by performing essential research leading to increased understanding of diseases, by conducting carefully controlled studies to test the effects of treatments on disease processes, and by directly administering (by neurosurgical, endovascular, endoscopic, or stereotactic means) the treatments to patients. PMID- 10807236 TI - Management of residual dysplastic vessels after cerebral arteriovenous malformation resection: implications for postoperative angiography. AB - OBJECTIVE: The verification of surgical resection of cerebral arteriovenous malformations (AVMs) relies on angiography. Abnormal vasculature often is identified after removal of the AVM. Differentiation of dysplastic feeding vessels that resemble the neovascularity of moyamoya disease, as distinct from residual AVM, is crucial for preserving critical brain areas. We review a large experience with immediate postoperative angiography after AVM resection and discuss the implications for management of abnormal dysplastic vessels discovered after AVM resection. METHODS: Beginning in 1992, 86 consecutive patients with AVMs underwent operations by standard protocol for immediate postoperative angiography under the same general anesthetic. Angiographic interpretation dictated admission to the intensive care unit or return to the operating room for further resection. RESULTS: In 78 patients, the angiogram revealed complete resection. Two patients were returned to the operating room, one for residual malformation with an early draining vein, and one for resection of residual dysplastic vessels. There was one postoperative hemorrhage in a patient whose postoperative angiogram was falsely negative for AVM. Six patients with residual dysplastic vessels mimicking residual AVM, but without an early draining vein, were managed conservatively. Delayed follow-up angiography demonstrated spontaneous involution of these abnormal vessels in all of these patients. CONCLUSION: Residual dysplastic feeding vessels resembling the neovascularity of moyamoya disease but not associated with an early draining vein do not necessarily represent residual malformation after AVM resection. The abnormal vessels will proceed to complete spontaneous resolution. Given the difficulty of interpreting intraoperative angiography, immediate postoperative angiography may be a viable alternative after AVM resection. PMID- 10807237 TI - Intracranial aneurysms and cocaine abuse: analysis of prognostic indicators. AB - OBJECTIVE: The outcome of subarachnoid hemorrhage associated with cocaine abuse is reportedly poor. However, no study in the literature has reported the use of a statistical model to analyze the variables that influence outcome. METHODS: A review of admissions during a 6-year period revealed 14 patients with cocaine related aneurysms. This group was compared with a control group of 135 patients with ruptured aneurysms and no history of cocaine abuse. Age at presentation, time of ictus after intoxication, Hunt and Hess grade of subarachnoid hemorrhage, size of the aneurysm, location of the aneurysm, and the Glasgow Outcome Scale score were assessed and compared. RESULTS: The patients in the study group were significantly younger than the patients in the control group (P < 0.002). In patients in the study group, all aneurysms were located in the anterior circulation. The majority of these aneurysms were smaller than those of the control group (8 +/- 6.08 mm versus 11 +/- 5.4 mm; P = 0.05). The differences in mortality and morbidity between the two groups were not significant. Hunt and Hess grade (P < 0.005) and age (P < 0.007) were significant predictors of outcome for the patients with cocaine-related aneurysms. CONCLUSION: Cocaine use predisposed aneurysmal rupture at a significantly earlier age and in much smaller aneurysms. Contrary to the published literature, this group did reasonably well with aggressive management. PMID- 10807238 TI - Basal brain injury in aneurysm surgery. AB - OBJECTIVE: The goal of this study was to determine the frequency of lesions in the basal frontotemporal area that were related to surgical damage to the brain tissue. METHODS: A prospective series of 101 patients with ruptured intracranial aneurysms were examined with high-field magnetic resonance imaging, 2 to 6 years (mean, 3.3 yr) after early surgery. RESULTS: Lesions in the basal frontotemporal region, on the side of the pterional approach, were observed for 36 patients. These lesions were not visible in computed tomographic scans obtained pre- or postoperatively or 3 months after subarachnoid hemorrhage. Patients with ruptured aneurysms in the anterior communicating artery exhibited fewer of these lesions than did patients with aneurysms in the internal carotid artery or middle cerebral artery; this difference was not statistically significant. The age of the patient, the duration and depth of hypotension, the amount of blood or ventricular enlargement in pre- and postoperative computed tomographic scans, and the incidence and severity of angiographic vasospasm in pre- and postoperative angiograms did not predict the existence of these lesions. The clinical conditions of the patients, as assessed using the Glasgow Outcome Scale, at 3 months after surgery and at the time of magnetic resonance imaging did not predict the existence of these lesions. Nine of the 10 patients who underwent surgical treatment of unruptured aneurysms on the contralateral side exhibited no signs of tissue damage. CONCLUSION: Surgical treatment of ruptured intracranial aneurysms seems to cause damage in the basal frontotemporal region in one-third of patients. The significance of these lesions remains unclear. PMID- 10807239 TI - A theory on the natural history of colloid cysts of the third ventricle. AB - OBJECTIVE: Patients with third ventricular colloid cysts typically are diagnosed when they develop symptoms related to cerebrospinal fluid (CSF) obstruction at the foramen of Monro. However, the clinical and neuroimaging characteristics related to symptom development are poorly understood. METHODS: From January 1974 to June 1998, 155 patients with newly diagnosed colloid cysts were managed at our center. Eighty-seven patients (56%) were thought to have tumor-related symptoms, and they underwent surgery (resection, n = 74; ventriculoperitoneal shunting, n = 11; stereotactic aspiration, n = 2). Sixty-eight patients (44%) had colloid cysts thought to be asymptomatic, and observation with serial neuroimaging was recommended. Recursive partitioning was used to examine the association of patient and tumor characteristics with the development of cyst-related symptoms. RESULTS: Univariate analysis comparing symptomatic and asymptomatic patients revealed four factors associated with cyst-related symptoms: 1) younger patient age (44 yr versus 57 yr; P < 0.001); 2) cyst size (13 mm versus 8 mm; P < 0.001); 3) ventricular dilation (83% versus 31%; P < 0.001); and 4) increased signal on T2-weighted magnetic resonance images (44% versus 8%; P = 0.001). All four variables remained significant in a multivariate logistic regression model: patient age (P = 0.04; odds ratio, 1.0); cyst size (P = 0.04; odds ratio, 1.2); ventricular dilation (P = 0.02, odds ratio, 7.2); and increased signal on T2 weighted magnetic resonance images (P = 0.04; odds ratio, 2.7). The most significant variable was ventriculomegaly (yes versus no). Patients with normal sized ventricles (n = 62) were further categorized by cyst size (< or = 10 mm versus > 10 mm). For patients with enlarged ventricles (n = 93), patient age (< or = 50 yr versus > 50 yr) was the most important variable. Patients older than 50 years also were split with respect to cyst size; patients aged 50 years or younger with enlarged ventricles were not affected by cyst size. The percentage of patients with cyst-related symptoms was 12, 50, and 85% in the three final patient classes, respectively. Multivariate analysis including the patient classes resulted in removal of the other significant variables from the model, whereas the patient classes remained significant (P < 0.0001; odds ratio, 6.3) for predicting patients with cyst-related symptoms. CONCLUSION: The patient and neuroimaging characteristics of the different patient classes support a theory on the natural history of colloid cysts. Patients with third ventricular colloid cysts become symptomatic when the tumor enlarges rapidly, causing CSF obstruction, ventriculomegaly, and increased intracranial pressure. Some cysts enlarge more gradually, however, allowing the patient to accommodate the enlarging mass without disruption of CSF flow, and the patient remains asymptomatic. In these cases, if the cyst stops growing, the patient can maintain a steady state between CSF production and absorption and may not require neurosurgical intervention. PMID- 10807240 TI - Primary malignant tumors of the sphenoidal sinus. AB - OBJECTIVE: Neoplasms of the paranasal sinuses account for 0.2 to 0.8% of all cancers and for 2 to 3% of head and neck cancers. Primary involvement of the sphenoidal sinus has been reported to occur in only 1 to 2% of all paranasal sinus tumors. Tumors at this site commonly present with cranial neuropathies and require multidisciplinary treatment. METHODS: During a 21-year period, 27 patients with primary sphenoidal sinus tumors were evaluated. Those with pituitary tumors and fibro-osseous lesions (fibrous dysplasia) were excluded. The records of all remaining patients were reviewed for the following parameters: age, sex, and ethnic origin; presenting symptoms and signs; radiological features and sites of tumor extension; pathological findings; prior treatments; surgical approaches and extent of resection; adjuvant therapy; complications; and patient outcome. RESULTS: A malignant pathological process was found in 26 of the 27 patients. The most common diseases were squamous cell carcinoma (n = 9), adenoid cystic carcinoma (n = 4), chondrosarcoma (n = 3), and neuroendocrine carcinoma (n = 3). Treatments included surgery (n = 1), surgery and radiotherapy (n = 6), surgery and chemotherapy (n = 3), surgery, radiotherapy, and chemotherapy (n = 4), chemotherapy (n = 5), chemotherapy and radiotherapy (n = 3), and radiotherapy (n = 2). The mean follow-up period from initial evaluation was 41 months (range, 2-199 mo). At the last follow-up, 12 patients (48%) were still alive and 13 (52%) had died. The 2-year survival rate for patients with squamous cell carcinoma was 44%. CONCLUSION: Headache and visual disturbances are common symptoms of sphenoidal sinus tumors. The presence of cranial neuropathies, although common, suggests a less successful outcome. It is advantageous if the surgical team has had experience with both open and endoscopic approaches to the sphenoidal sinus. Patient outcome is maximized by the use of aggressive multidisciplinary therapy. PMID- 10807241 TI - A prospective population-based study of pediatric trauma patients with mild alterations in consciousness (Glasgow Coma Scale score of 13-14). AB - OBJECTIVE: Considerable controversy surrounds the appropriate evaluation of children with mild alterations in consciousness after closed head trauma (Glasgow Coma Scale [GCS] score of 13-14). The objective of the current study was to determine the incidence of intracranial lesions in pediatric patients with a field GCS score of 13 or 14 after closed head injuries. METHODS: The current study is a population-based, multicenter prospective study of all patients to whom emergency medical services responded during a 12-month period. The setting was urban Los Angeles County, encompassing a patient population of 2.3 million children, 13 designated trauma centers, and 94 receiving hospitals. RESULTS: In the pediatric age group (<15 yr old), 8488 patients were transported by emergency medical services for injuries. Of these, 209 had a documented field GCS score of 13 or 14. One hundred fifty-seven patients were taken to trauma centers, and 135 (86%) underwent computed tomography. Forty-three patients (27.4%) had abnormal results on computed tomographic scans, 30 (19.1%) had an intracranial hemorrhage, and 5 required an operative neurosurgical procedure for hematoma evacuation. Positive and negative predictive values of deteriorating mental status (0.500 and 0.844, respectively), loss of consciousness (0.173 and 0.809), cranial fracture (0.483 and 0.875, and extracranial injuries (0.205 and 0.814) were poor predictors of intracranial hemorrhage. CONCLUSION: Pediatric patients who have mild alterations in consciousness in the field have a significant incidence of intracranial injury. The great majority of these patients will not require operative intervention, but the implications of missing these hemorrhages can be severe for this subgroup of head-injured patients. Because clinical criteria and cranial x-rays are poor predictors of intracranial hemorrhage, it is recommended that all children with a GCS score of 13 or 14 routinely undergo screening via non-contrast-enhanced computed tomography. PMID- 10807242 TI - Risk factors for failure of endoscopic third ventriculostomy for obstructive hydrocephalus. AB - OBJECTIVE: This is a retrospective study to identify risk factors for failure in the treatment of obstructive hydrocephalus with endoscopic third ventriculostomy (ETV). METHODS: The records for 89 patients, including 32 with ventriculoperitoneal or ventriculoatrial shunt malfunctions or infections, who underwent ETVs between 1993 and 1998, at our institution, were examined. Multiple variables possibly related to failure were considered. These included age, sex, cause of hydrocephalus, presence and function of ventriculoperitoneal/ventriculoatrial shunts, history of shunt revisions or infections, symptoms, preoperative imaging results, presence of retained shunt catheters, postoperative meningitis, and postoperative ventricular size. RESULTS: Twenty-nine patients (32.6%) required subsequent shunt replacement and/or ETV revision. Of these 29 reoperations, 12 procedures (41.4%) were performed within 2 weeks and only 3 were performed more than 10 months after the initial ETV procedure. The ventricular size remained unchanged in 75% of the cases on the day after ETV, in 57.4% at 3 months, in 48.2% at 6 months, and in 41.8% at 1 year. Cine phase-contrast magnetic resonance imaging findings were consistent with postoperative symptomatic resolution in 96.3% of the cases. Seven patients (7.9%) experienced complications related to ETV, all of which were transient. Significant risk factors in univariate analyses were as follows: presence of Chiari Type I malformation (P = 0.003), shunt infection at presentation (P = 0.014), history of shunt infections (P = 0.0004), three or more previous shunt revisions (P = 0.0018), and postoperative meningitis (P = 0.0001). Late-onset idiopathic aqueductal stenosis was a significant predictor of good outcomes (P = 0.044). These factors were reanalyzed in a multivariate analysis, which confirmed a history of shunt infections and postoperative meningitis as independent risk factors. CONCLUSION: The risk of failure increases with intracerebral infection, likely because of obliteration of cerebrospinal fluid pathways. PMID- 10807243 TI - Clinical evaluation and follow-up results for intraoperative magnetic resonance imaging in neurosurgery. AB - OBJECTIVE: The use of intraoperative magnetic resonance imaging (MRI) in neurosurgery has increased rapidly, and a variety of concepts have recently been presented. Although the feasibility of the procedure has been demonstrated repeatedly, no conclusive analysis of its effects on the surgical procedures, the extent of tumor removal, and outcomes, or its possible problems, has been performed. METHODS: Of 242 operations performed with intraoperative MRI, 97 procedures for supratentorial glioma treatment were analyzed with respect to intraoperative imaging results and postoperative outcomes. Analysis of the images included assessment of imaging artifacts, image quality, and extent of tumor removal. Patients were monitored to determine radiological progression, survival times, postoperative complications, and morbidity rates. RESULTS: No intraoperative complications related to the imaging procedure were observed. Image quality was good or fair in 85.5% of the cases. Different types of surgically induced imaging changes could be identified. In 56 cases, resection was continued using navigation with intraoperative MRI data sets (rereferencing accuracy, 0.9 mm). For high-grade gliomas, the percentage of cases in which residual tumor was identified by MRI could be significantly reduced from 62% intraoperatively to 33% postoperatively, which was paralleled by a significant increase in survival times for patients without residual tumor. Complication and morbidity rates were within the ranges reported for other studies. CONCLUSION: Intraoperative MRI is safe and allows reliable updating of neuronavigational data, with compensation for brain shifting. Surgically induced imaging changes, which have been identified as a possible problem with intraoperative MRI in general, necessitated comparisons with preoperative scans and require future attention. The extent of tumor removal and survival times were increased significantly. Overall, patients seemed to benefit from the method. PMID- 10807244 TI - Permanent iodine-125 interstitial implants for the treatment of recurrent glioblastoma multiforme. AB - OBJECTIVE: Brachytherapy with temporary implants may prolong survival in patients with recurrent glioblastoma multiforme (GBM), but it is associated with relatively high costs and morbidity. This study reports the time to progression and survival after permanent implantation of iodine-125 seeds for recurrent GBM and examines factors predictive of outcome. METHODS: Forty patients with recurrent GBM were treated with maximal resection plus permanent placement of iodine-125 seeds into the tumor bed. A total dose of 120 to 160 Gy was administered, and patients were followed up with magnetic resonance imaging scans every 2 to 3 months. RESULTS: Actuarial survival from the time of implantation was 47 weeks, with 7 of 40 patients still alive at a median of 59 weeks after implantation. Survival was significantly better for patients younger than 60 years, and a trend for longer survival was demonstrated with gross total resection and tumors with a low MIB-1 (a nuclear antigen present in all cell cycles of proliferating cells) staining index. Median time to progression was 25 weeks and, on multivariate analysis, was favorably influenced by gross total resection and patient age younger than 60 years. After implantation, 27 of 30 patients with failure had a local component to the failure. No patient developed symptoms attributable to radiation necrosis or injury. CONCLUSION: Permanent iodine-125 implants for recurrent GBM result in survival comparable with that described in previous reports on temporary implants, but with less morbidity. Results are most favorable for patients who are younger than 60 years, and who undergo gross total resection. Despite this aggressive treatment, most patients die as a consequence of locally recurrent disease. PMID- 10807245 TI - Limits of intermittent jugular bulb oxygen saturation monitoring in the management of severe head trauma patients. AB - OBJECTIVE: To evaluate, in a prospective, observational study, whether bilateral monitoring of jugular bulb oxyhemoglobin saturation (SjO2), in addition to standard monitoring, results in modification of the management of severe head trauma. METHODS: The patients underwent bilateral jugular bulb cannulation and observation at 8-hour intervals, during which SjO2 was measured and the neurological condition and physiological variables were assessed. The study group was responsible for evaluating whether the physician's decision-making process was influenced by the detection of SjO2 abnormalities. The SjO2 discrepancy in simultaneous bilateral samples was also evaluated to determine whether it interfered with the interpretation of data and with clinical decision-making. The SjO2-related complications were monitored. RESULTS: Thirty patients underwent 319 observations. In 96% of patients, SjO2 was normal or high and had no influence on the diagnostic or therapeutic strategies. Treatment decisions were dictated by changes in clinical status and in intracranial and cerebral perfusion pressure. When these parameters were abnormal, treatment was administered, even if SjO2 was normal (101 observations). Conversely, when SjO2 was the only detected abnormality (34 observations), no treatment was administered. Abnormally low SjO2 values, caused by hypovolemia and hypocapnia, were detected in 3.4% of observations and actually modified the management. The discrepancies in simultaneous bilateral samples were substantial and gave rise to relevant interpretation problems. Fifteen percent of jugular catheters showed evidence of bacterial colonization. CONCLUSION: Intermittent SjO2 monitoring did not substantially influence the management of severe head trauma. Therefore, recommendation for its routine use in all patients seems inadvisable, and indications for this invasive method should no longer be defined on the basis of experts' opinions, but rather on randomized, prospective studies. PMID- 10807246 TI - Quantitative parameters of intraoperative electromyography predict facial nerve outcomes for vestibular schwannoma surgery. AB - OBJECTIVE: Facial nerve monitoring is an established method that is routinely used during cerebellopontine angle tumor surgery. The aim of this study was to determine quantitative electromyographic (EMG) parameters that were predictive of facial nerve outcomes. METHODS: In 137 patients with intra-/extrameatal vestibular schwannomas, the most proximal (the exit from the brainstem) and distal (the fundus of the internal auditory canal) parts of the facial nerve were stimulated after total tumor removal. A quantitative analysis of absolute values and ratios (proximal/distal) of evoked EMG parameters (amplitude, latency, and duration) was performed, and parameters were correlated with postoperative (1 and 6 wk and 6 mo) facial nerve function (FNF). RESULTS: Absolute values of EMG amplitudes were statistically correlated with FNF (P < 0.05). Amplitude ratios (proximal/distal) demonstrated an even greater predictive power. The risk of exhibiting facial palsy 6 months after surgery increased from 1.6% (amplitude ratio of >0.8) to 75% (ratio of <0.1). For EMG latencies, only the ratios revealed a significant correlation with FNF. The latency ratio-dependent risk of facial palsy after 6 months increased from 2.9% (ratio of <1.05) to 33% (ratio of >1.35). The durations of the muscle responses were not significantly correlated with clinical outcomes. CONCLUSION: The predictive power of the amplitudes and latencies of electrically evoked muscle responses could be improved by calculating proximal/distal ratios. The proximal/distal amplitude ratio proved to be the most powerful parameter for intraoperative assessment of postoperative FNF. PMID- 10807247 TI - The value of routine cultures of the cerebrospinal fluid in patients with external ventricular drains. AB - OBJECTIVE: The purpose of this study was to determine whether routine cerebrospinal fluid (CSF) bacteriological cultures in patients with external ventricular drains (EVDs) can identify infections early and prevent complications related to bacterial ventriculitis. METHODS: We retrospectively reviewed the microbiological reports and clinical data for all patients in whom an EVD was placed at a tertiary care pediatric neurosurgical center between 1984 and 1997. EVDs were inserted in the operating room or intensive care unit, and, in most patients whose EVD remained in place for more than 2 days, daily cultures of CSF were performed. RESULTS: One hundred fifty-seven patients in whom 160 EVDs had been placed were included in the study. Forty-eight positive cultures were identified, of which the majority were determined to be contaminants. Seven infections were identified on the basis of microbiological criteria (i.e., a gram positive stain and positive culture) and a subsequent positive culture. In all patients in whom infections developed, routine daily cultures of CSF were performed, and, in each instance, these cultures failed to identify the infections before clinical changes occurred. All seven patients with infection had fever (>38.5 degrees C) and peripheral leukocytosis (>11 x 10(3)/mm3) on the day the infection was identified, and one had a change in CSF appearance. CONCLUSION: The results of this study suggest that routine culture of CSF in children with EVDs is not necessary, and that if CSF cultures are performed for new fever (>38.5 degrees C) or peripheral leukocytosis, neurological deterioration, or a change in CSF appearance, infections will be identified in a timely fashion. In situations in which these clinical indicators might be masked, routine cultures may be valuable. PMID- 10807248 TI - Contralateral approach to junctional C2-C3 and proximal C4 aneurysms of the internal carotid artery: microsurgical anatomic study. AB - OBJECTIVE: To evaluate a contralateral approach to aneurysms located in the internal carotid artery cave and proximal C4 segments. METHODS: In six adult cadaveric head sides, proposed aneurysms in the carotid cave or proximal C4 segments were approached via contralateral craniotomies. We summarize the approach in the following steps: 1) frontotemporal orbital craniotomy, 2) drilling of the lateral sphenoid wing and opening of the dura along the frontotemporal base, 3) drilling of the planum sphenoidale and the tuberculum sellae more extensively toward the aneurysm side and opening of the sphenoid sinus, 4) drilling of the medial part of the anterior clinoid process on the side of the aneurysm and removal of the superior, medial, and inferior walls of the optic canal, 5) opening of the optic sleeve, and 6) opening of the space between the medial wall of the internal carotid artery C2-C3 segments and the lateral edge of the pituitary gland. RESULTS: The contralateral approach to expose the opposite internal carotid artery cave and proximal C4 segments provided excellent views of the region, without mobilization or retraction of either the optic nerve or the carotid artery. CONCLUSION: We recommend that this approach be used only for selected aneurysms, which are small and directed medially, anteriorly, or inferiorly, in the defined locations. PMID- 10807249 TI - Thermological study of drilling bone tissue with a high-speed drill. AB - OBJECTIVE: To perform a detailed quantitative analysis of the effect of intermittent drilling and irrigation to lower the temperature during high-speed drilling. METHODS: We examined the thermal changes for 15 bone flaps while drilling a 7- x 7-mm area for 18 seconds and a specific point for 9 seconds, under the following conditions: continuous or intermittent drilling; with a fluted steel or a diamond ball cutter; without or with irrigation, with room temperature or cold (8 degrees C) Ringer solution. RESULTS: The maximum temperature during continuous drilling with a diamond cutter (or steel cutter) was 82.4 +/- 1.3 degrees C (55.2 +/- 1.0 degrees C) without irrigation, 42.5 +/- 1.2 degrees C (35.4 +/- 0.9 degrees C) with room temperature Ringer irrigation, and 22.5 +/- 1.4 degrees C (21.6 +/- 0.3 degrees C) with cold Ringer irrigation, whereas that during intermittent drilling was 66.1 +/- 1.2 degrees C (35.6 +/- 0.8 degrees C), 35.0 +/- 0.8 degrees C (25.4 +/- 0.7 degrees C), and 21.5 +/- 0.6 degrees C (21.8 +/- 0.4 degrees C), respectively. During continuous drilling of a specific point, the radius of the concentric isothermal line for 43 degrees C (thermal threshold for neurons) was 3.6 +/- 0.8 mm with a steel cutter, and it was 8.1 +/- 0.3 mm with a diamond cutter. The radius was 4.2 +/- 0.8 mm during intermittent drilling with a diamond cutter, and it was 4.0 +/- 0.1 mm during continuous drilling with room temperature Ringer irrigation. Intermittent drilling with cold Ringer irrigation kept the temperature, even at the center of the drilled area, below 20 degrees C. CONCLUSION: Intentional intermittent drilling with irrigation reduces temperature elevation and its expansion. These procedures are necessary for safe drilling, especially with a diamond burr. Although cold irrigation can minimize temperature elevation, its substantial effect on nerves or other structures needs to be elucidated. PMID- 10807250 TI - Human glioma immunobiology in vitro: implications for immunogene therapy. AB - OBJECTIVE: Human gliomas are known to be immunosuppressive. Recent reports have suggested novel strategies to overcome this immunosuppression, including immunogene therapy. We examined expression of 10 immunologically important molecules by human gliomas in vitro, and we discuss the implications for immunogene therapy. METHODS: Early passage human glioma cultures and established human glioma cell lines were analyzed by flow cytometry for expression of Class I and II major histocompatibility complex (MHC), B7-2 (CD86), and Fas (CD95). Culture supernatants were assayed by enzyme-linked immunosorbent assay for interleukin (IL)-6, IL-10, IL-12, transforming growth factor beta2, prostaglandin E2, and granulocyte-macrophage colony-stimulating factor levels. RESULTS: All cultures (16 of 16 samples) expressed Class I MHC and Fas, but few expressed Class II MHC (1 of 16 samples) or B7-2 (0 of 16 samples). Nearly all expressed high levels of IL-6 (19 of 21 samples; mean, 36.5 +/- 10.8 ng/10(6) cells/d) and prostaglandin E2 (21 of 21 samples; mean, 15.6 +/- 4.5 ng/10(6) cells/d) levels, and many expressed transforming growth factor beta2 (13 of 21 samples; mean, 8.6 +/- 3.7 ng/10(6) cells/d). Although several cultures (6 of 14 samples) expressed granulocyte-macrophage colony-stimulating factor, expression levels were very low (mean, 0.2 +/- 0.1 ng/10(6) cells/d). Few cultures (4 of 21 samples) expressed measurable IL-10, and none (0 of 22 samples) expressed IL-12. CONCLUSION: Class I MHC and Fas expression suggests that human glioma cells may be susceptible to Class I MHC-dependent cytotoxic T cell recognition and Fas-mediated killing. Unfortunately, transforming growth factor beta2 and prostaglandin E2 probably impair T cell activation, and IL-6 may shift immunity to less effective humoral (T helper 2) responses. Proinflammatory gene expression (B7-2, granulocyte macrophage colony-stimulating factor, and/or IL-12) is lacking. Together, these results suggest that modifying glioma cells via proinflammatory gene transfer or immunoinhibitory gene suppression might stimulate immune responses that are effective against unmodified tumors. PMID- 10807251 TI - Expression of structural proteins and angiogenic factors in cerebrovascular anomalies. AB - OBJECTIVE: The goal of this study was to describe the expression of matrix proteins and angiogenic factors in cerebrovascular malformations. METHODS: Forty six cerebrovascular malformations were immunohistochemically investigated with a battery of staining for five structural proteins (collagen IV, collagen III, smooth muscle actin, fibronectin, and laminin), and three angiogenic factors (vascular endothelial growth factor [VEGF], basic fibroblast growth factor [bFGF], and transforming growth factor alpha [TGFalpha]). The lesions consisted of 34 arteriovenous malformations (AVMs), 10 cavernous malformations (CMs), and 2 venous angiomas. Expression intensity for each histological layer in the abnormal vessel wall was graded and compared. RESULTS: AVM endothelia and subendothelia expressed more laminin and collagen IV than the same layers of CMs. Conversely, CMs expressed more fibronectin than AVMs. CM endothelia exhibited more prominent staining for smooth muscle actin than AVM endothelia. AVMs and CMs expressed VEGF in the endothelium and subendothelium, and TGFalpha in endothelial and perivascular layers. However, unlike AVMs, CMs expressed bFGF in the endothelium as well. The brain tissue intermingled within AVMs also expressed growth factors. Modified glial cells in the brain tissue adjacent to CMs expressed bFGF and TGFalpha, but not VEGF. Venous angiomas did not express the studied growth factors and mainly consisted of structural proteins of angiogenically mature tissue. CONCLUSION: Expression characteristics of structural proteins reveal that AVMs and CMs have different immunohistological properties. This study provides strong confirmation of previous findings of VEGF and bFGF immunoexpression in AVMs and CMs. It adds new information on TGFalpha expression in these malformations and on expression of the angiogenic factors in venous angiomas. PMID- 10807252 TI - Effect of adenovirus-mediated nitric oxide synthase gene transfer on vasospasm after experimental subarachnoid hemorrhage. AB - OBJECTIVE: Evidence indicates that vasospasm after subarachnoid hemorrhage (SAH) is caused in part by a decrease in the vasodilator nitric oxide (NO), which is produced mainly in endothelial cells. This study tested whether intracisternal injection of adenovirus-expressing endothelial NO synthase (eNOS) would decrease vasospasm in dogs. METHODS: In 12 dogs, baseline cerebral angiography was performed, and then SAH was produced by two injections of blood into the cisterna magna. The dogs were randomized (n = 6/group) to intracisternal injection of adenovirus-expressing lacZ (Ad327beta-Gal) or eNOS (AdCD8-NOS), administered immediately after the first blood injection. Angiography was repeated on Day 7, and then L-arginine (50 mg) was administered intracisternally, and angiography was repeated. Cerebrospinal fluid aspirated from the cisterna magna on Days 2 and 7 was analyzed for levels of NO metabolites. The dogs were killed, and their basilar arteries were removed and studied pharmacologically. Four control dogs underwent angiography on Day 0, followed by virus injection (n = 2/group). Angiography was repeated on Day 7, and the control dogs were killed. Transgene expression was detected in tissue removed on Day 7 by histochemical staining for lacZ, by polymerase chain reaction for messenger ribonucleic acid for eNOS, and by measurement of NO metabolites in cerebrospinal fluid. RESULTS: Angiography showed significant vasospasm in each group (Ad327beta-Gal, -54 +/- 7% reduction in basilar artery diameter; AdCD8-NOS, -53 +/- 7%), with no significant difference between groups. Injection of L-arginine caused an insignificant increase in arterial diameter in each group. In dogs without SAH, Ad327beta-Gal caused a reduction in basilar artery diameter (-13 +/- 10%, P = 0.42; paired t test), whereas injection of AdCD8-NOS caused an increase in diameter (14 +/- 16%, P = 0.77; paired t test). Histological examination and beta-galactosidase staining of dogs given injections of Ad327beta-Gal showed staining in inflammatory cells in the subarachnoid space, in the adventitia of the cerebral vessels, and in the liver and lungs. Messenger ribonucleic acid for eNOS was detected in the leptomeninges of dogs given injections of AdCD8-NOS. Under isometric tension, basilar arteries from each group demonstrated similar relaxation to L-arginine, but arteries exposed to eNOS demonstrated significantly greater relaxation to L-arginine plus tetrahydrobiopterin than arteries exposed to lacZ. Cerebrospinal fluid levels of NO and its metabolites were significantly higher in dogs treated with AdCD8-NOS than those treated with Ad327beta-Gal 2 days after SAH. CONCLUSION: These results demonstrate that adenovirus vectors can be used to transfer genes to cells in the subarachnoid space of dogs. Enough NO can be produced in the absence of SAH to dilate the basilar artery. After SAH, however, NO plus a cofactor can dilate arteries in vitro, but not enough NO is generated in the subarachnoid space to prevent vasospasm, perhaps owing to the scavenging of NO by hemoglobin. PMID- 10807253 TI - Significance of resting and stimulated cerebral blood flow for predicting the risk of hemodynamic cerebral ischemia in a model of chronic hemodynamic insufficiency. AB - OBJECTIVE: It has been postulated that patients with a compromised cerebrovascular reserve capacity (RC), defined as cerebral blood flow (CBF) response to acetazolamide (ACZ) (by percent), are at higher risk for ischemic stroke. The value of CBF and RC for predicting the risk of hemodynamically induced impairment of cerebral function is examined. METHODS: Both common carotid arteries were occluded in 22 Wistar-Kyoto rats. Thirty-one days later, mean arterial blood pressure was reduced to 40 mm Hg for 30 minutes. Laser Doppler scanning of CBF at resting conditions and after intraperitoneal administration of ACZ (0.1 mg/g body weight) was performed 30 minutes and 28 days after occlusion as well as before and during hypotension. Memory and motor functions were examined before and after CBF measurements. RESULTS: After occlusion, CBF dropped significantly and ACZ did not increase CBF. Four weeks later, resting CBF had significantly improved but remained impaired, as did RC, showing a distinct interindividual variability. Hypotension reduced CBF by 57 +/- 4% (P < 0.001) and significantly impaired memory and motor functions. CBF during hypotension correlated with resting CBF before hypotension (r = 0.495, P = 0.027) and with CBF before (r = 0.392, P = 0.048) and after (r = 0.476, P = 0.034) ACZ, as determined 4 weeks after occlusion. There was no correlation with RC (r = 0.091, P = 0.702). Neurological tests performed 1 day after hypotension correlated with CBF during hypotension (memory function, P = 0.03; motor function, P = 0.02) but not with RC. CONCLUSION: In this model of chronic hemodynamic insufficiency, the risk of impairment to global cerebral function was predicted by resting CBF and CBF after ACZ but not by RC determined with ACZ. PMID- 10807254 TI - Bone morphogenetic proteins and bone morphogenetic protein gene therapy in neurological surgery: a review. AB - OBJECTIVE: To review the uses of bone morphogenetic proteins (BMPs) and BMP gene therapy for the treatment of neurosurgical disorders. METHODS: Literature review. RESULTS: BMPs are members of the transforming growth factor beta superfamily, and they play an important role in the growth and development of numerous tissues, including bone, brain, and spinal cord. Although the majority of previous studies have focused on the regulatory functions of BMPs in the normal growth and differentiation of the skeletal system, BMPs also seem to be exquisitely involved in the regulation of cellular proliferation, survival, differentiation, apoptosis, and lineage commitment in the central nervous system. When specific BMPs are delivered on biological matrices, they have the capacity to induce bone, cartilage, ligament, and tendon at both heterotopic and orthotopic sites, suggesting that they may play a major role in the future treatment of spinal and craniofacial pathology. For example, recent studies have clearly demonstrated the usefulness of BMPs and BMP gene therapy for the induction of spinal arthrodesis in several animal models. In addition, several BMPs have been shown to have a neuroprotective effect in animal models of head injury, cerebral ischemia, and Parkinson's disease and may therefore have direct clinical applications for the treatment of central nervous system disorders. CONCLUSION: As the physiological activity of BMPs in the development and pathology of the central nervous system and spine are more fully elucidated, BMP therapeutics and gene therapy will probably have numerous applications in neurological surgery. PMID- 10807255 TI - Neurosurgery at the University of Pennsylvania Medical Center. AB - The University of Pennsylvania Medical School was the nation's first medical school, and its Department of Neurosurgery is one of the nation's oldest. The history of the Department of Neurosurgery at Penn is recounted, beginning with the pioneer surgeon Charles Harrison Frazier. The evolution of the current department, its contemporary status, and its residency program are described. PMID- 10807256 TI - Symptomatic lateral ventricular ependymal cysts: criteria for distinguishing these rare cysts from other symptomatic cysts of the ventricles: case report. AB - OBJECTIVE AND IMPORTANCE: Symptomatic lateral ventricular ependymal cysts are rare. Two previous cases of this lesion have been reported in the literature. We report a third case and provide radiological and histopathological criteria for differentiating this entity from common intracranial cysts. CLINICAL PRESENTATION: A 43-year-old man presented with a 6-year history of seizures and progressive right occipitoparietal headaches. Computed tomography and magnetic resonance imaging demonstrated a 4- x 3- x 3-cm nonenhancing cystic mass, expanding the trigone of the right lateral ventricle. INTERVENTION: The patient underwent a right occipital craniotomy. The cyst was opened, fluid was aspirated, the cyst wall was biopsied, and a cyst-subarachnoid communication was established. The patient did well postoperatively, with no seizures and with resolution of headaches. CONCLUSION: Lateral ventricular ependymal cysts are a rare cause of neurological symptoms, including headache and seizure. Distinctive radiographic characteristics distinguish these cysts at preoperative evaluation. Careful analysis of the histopathology and immunohistochemistry studies correctly identifies these lesions, gives insight into the natural history of ependymal cysts, and guides clinical management decisions. PMID- 10807257 TI - Pituitary carcinoma containing gonadotropins: treatment by radical excision and cytotoxic chemotherapy: case report. AB - OBJECTIVE AND IMPORTANCE: Pituitary carcinomas are extremely rare. Cases reported in the medical literature in the 20th century included tumors that produced adrenocorticotropic hormone, prolactin, growth hormone, and/or thyrotropin. CLINICAL PRESENTATION: Here we present a 22-year-old woman with a pituitary carcinoma that was immunohistochemically positive for luteinizing hormone and follicle-stimulating hormone at both the primary and metastatic sites. The patient exhibited elevated serum levels of alpha-subunit. INTERVENTION: The patient had experienced failure of previous treatments, including standard surgery and radiotherapy, and presented to us for radical resection of the tumor, with exenteration of the involved cavernous sinus. She was pretreated with cytotoxic chemotherapy and continued to receive this therapy after surgery. CONCLUSION: This is the only documented case of a gonadotropin-staining pituitary carcinoma for which hormone production was proven in both the primary and metastatic tumors. Many benign "nonsecreting" pituitary adenomas actually produce subclinical amounts of gonadotropins, and malignant nonfunctional pituitary neoplasms may do the same. PMID- 10807258 TI - Type IV spinal arteriovenous malformation in association with familial pulmonary vascular malformations: case report. AB - OBJECTIVE AND IMPORTANCE: Type IVc arteriovenous malformations (AVMs) of the spinal cord consist of multiple high-flow feeding vessels, and they often present a challenging management situation. Their location is intradural and extramedullary, and they are rare malformations that are difficult to treat owing to the risk of thrombosis of the anterior spinal artery. The authors report a case of Type IVc spinal AVM in a patient with a family history of three siblings with pulmonary AVMs. Spinal AVMs have been reported to be associated with inherited syndromes such as familial cutaneous hemangiomas and Kartagener's syndrome, but an association with pulmonary AVMs has not previously been described. CLINICAL PRESENTATION: A 27-year-old man presented with sudden onset of occipital headache with cervical radiation while weightlifting. Results of computed tomography of the brain were normal, but lumbar puncture revealed a subarachnoid hemorrhage. The patient had a 1-year history of a neurogenic bladder and exhibited marked left calf muscle wasting. INTERVENTION: The patient underwent spinal magnetic resonance imaging, which revealed the AVM in the conus region. Selective spinal angiography was performed for diagnostic purposes. A laminectomy was performed, and the vessels feeding the AVM were clipped, as was the fistula. CONCLUSION: The patient remained neurologically stable, and angiography confirmed obliteration of the AVM. This is the first case report of a patient with a spinal AVM who had multiple siblings with pulmonary malformations or AVMs. PMID- 10807259 TI - Giant intraspinal pseudomeningoceles cause delayed neurological dysfunction after brachial plexus injury: report of three cases. AB - OBJECTIVE AND IMPORTANCE: Delayed neurological dysfunction after a brachial plexus injury is uncommon. We present the cases of three patients with a history of significant brachial plexus trauma and late neurological deterioration secondary to giant intraspinal extradural pseudomeningoceles. CLINICAL PRESENTATION: Three patients, each with a remote history of brachial plexus trauma, presented with slowly progressive upper-limb weakness. An examination revealed bilateral lower motor neuron weakness in the upper extremities in all patients and evidence of spastic paraparesis in one. Magnetic resonance imaging and postmyelogram computed tomographic scans demonstrated large anterior extradural cerebrospinal fluid collections extending from the upper cervical to lower thoracic and lumbar levels in each patient. Myelograms demonstrated a connection with the subarachnoid space in two patients. INTERVENTION: Direct obliteration of the connection between the cyst and the subarachnoid space was completed in two patients, and a cystoperitoneal shunt was placed in the third. Postoperative imaging demonstrated complete resolution of the extradural collections. Arrest of progression of upper-limb deterioration was observed in all patients, and dramatic improvement of long tract symptoms occurred in one. CONCLUSION: Giant intraspinal pseudomeningoceles are a rare complication of brachial plexus root injuries or avulsion, capable of causing significant morbidity. Early intervention can improve symptoms related to long tract involvement and prevent further deterioration of lower motor neuron disease. The pathophysiology of neurological dysfunction caused by these giant collections is unclear; however, vascular and mechanical factors thought to be important in the pathogenesis of cervical myelopathy also may have a role. PMID- 10807260 TI - Scalp marking for a craniotomy using a laser pointer during preoperative computed tomographic imaging: technical note. AB - OBJECTIVE: The preoperative localization of superficial intracranial lesions is often necessary for accurate burr hole placement or craniotomy siting. It is not always easy, however, to localize the lesions over the scalp working only from computed tomographic images. We developed a simple method for such localization using a laser pointer during the preoperative computed tomographic examination. METHODS: The angle of incidence, extending from a point on the scalp to the center of the computed tomographic image, is measured by the software included with the scanner. In the gantry, at the same angle as on the image, a laser is beamed from a handmade projector onto the patient's scalp toward the center of the gantry. The point illuminated on the patient's head corresponds to that on the image. The device and the method are described in detail herein. RESULTS: We applied this technique to mark the area for the craniotomy before surgery in five patients with superficial brain tumors. At the time of surgery, it was confirmed that the tumors were circumscribed precisely. CONCLUSION: The technique is easy to perform and useful in the preoperative planning for a craniotomy. In addition, the device is easily constructed and inexpensive. PMID- 10807261 TI - Scaled clips for aneurysm surgery: technical note. AB - OBJECTIVE: The tips of clip blades are often invisible during operations on large, deep, or posteriorly projecting aneurysms. To overcome this problem, we sought to develop a series of scaled clips. METHODS: We made scaled clips modified from Yasargil titanium clips. The blades of the new clips are scaled every 5 mm from the tips with alternating colors. We have added six new types of straight scaled clips. RESULTS: To date, we have performed approximately 40 aneurysm clippings using the new scaled clips and have confirmed their usefulness. CONCLUSION: Scaling on the blades helps confirm accurate placement of a clip, even during operations for large, deep, or posteriorly projecting aneurysms, in which blade tips are often invisible. PMID- 10807262 TI - Microinvasive transaxillary thoracoscopic sympathectomy: technical note. AB - OBJECTIVE: To describe a two-port transaxillary thoracoscopic approach for thoracic sympathectomy that maximizes working space, improves manipulative ability, and enhances visualization of the surgical field. METHODS: Positioning of the patients was optimized to displace the scapula posteriorly, widen the avenue of approach to the sympathetic ganglia, and create a more direct route to the target. The semi-Fowler position permitted the lung apex to fall away from mediastinal structures, obviating a separate retraction port. A 30-degree endoscope allowed an unobstructed view of surgical progress, and anatomic relationships were manipulated in a temporal sequence to facilitate dissection. RESULTS: Microinvasive transaxillary sympathectomy was performed successfully in 13 patients, all of whom had a good outcome without complications. CONCLUSION: The modifications implemented increase the speed and safety of thoracoscopic sympathectomy while minimizing complications. PMID- 10807263 TI - Freely movable armrest for microneurosurgery: technical note. AB - OBJECTIVE: To introduce an armrest that can move freely and can promptly be fixed at any desired position during surgery. METHODS: We developed a surgical armrest system that incorporates a mechanical arm, equipped with a ball joint, which can be fixed by frictional resistance. The mechanical arm moves freely as compressed air is applied. As the button switch below the armrest is pressed while the armrest is being held (grip switch), the mechanical arm can move freely to the desired position. The mechanical arm is locked in position as soon as the button is released. RESULTS: The stability of the surgeon's hands was markedly increased, and fatigue was markedly decreased, during surgery conducted using this armrest system. The system was used for 60 operations in 6 months without any problems. CONCLUSION: The freely movable armrest is useful for neurosurgical procedures that require agility and fine movements of the hand. PMID- 10807264 TI - Library: historical perspective. Ernest Sachs (1879-1958). PMID- 10807265 TI - Lymphocytic and granulomatous hypophysitis: experience with nine cases. PMID- 10807266 TI - Subtemporal decompression: the treatment of noncompliant ventricle syndrome. PMID- 10807267 TI - Arctic and Antarctic exploration including the contributions of physicians and effects of disease in the polar regions. PMID- 10807268 TI - Frameless stereotaxy for transsphenoidal surgery. PMID- 10807269 TI - Craniotomy for tumor treatment in an intraoperative magnetic resonance imaging unit. PMID- 10807270 TI - The development of anatomic art and sciences: the Ceroplastica anatomic models of La Specola. PMID- 10807271 TI - Combined laminectomy and thoracoscopic resection of a dumbbell neurofibroma: technical case report. PMID- 10807272 TI - Mutations in KRIT1 in familial cerebral cavernous malformations. AB - OBJECTIVES: The recognition of six unrelated Hispanic-American families in which cerebral cavernous malformations (CCM) segregated as an autosomal dominant trait established a genetic basis for this disease. Linkage analysis subsequently identified locus heterogeneity with disease genes for CCM at chromosomal regions 7q, 7p, and 3q. Recently, mutations in KRIT1, a gene on 7q at the CCM1 locus, were identified in French and Hispanic-American families with CCM. This study confirms the identity the KRIT1 founder mutation in Hispanic-Americans and reports a novel KRIT1 mutation in a Caucasian family. METHODS: Oligonucleotide primers were designed to allow amplification of genomic DNA sequences from four Hispanic-American families and five non-Hispanic families for all 12 exons of the KRIT1 gene using the polymerase chain reaction (PCR). The amplified DNA was then screened using single strand conformation polymorphism analysis (SSCP) and sequencing. The expression pattern of KRIT1 was analyzed by Northern blotting. RESULTS: Analysis of the KRIT1 gene revealed a point mutation in exon 6 that predicts the substitution of a premature termination codon for glutamine at codon 248 in all four Hispanic-American families, confirming previous findings. SSCP analysis and sequencing revealed an 11 base pair duplication in exon 7 leading to a premature termination codon in one Caucasian family. Northern analysis demonstrated widespread expression of this gene, however, the highest level of expression was in the brain. CONCLUSION: The common KRIT1 mutation causing the majority of CCM in Hispanic-Americans has been identified and independently confirmed, allowing efficient presymptomatic molecular diagnosis. In keeping with prior results, both newly identified mutations create a premature termination codon and are predicted to initiate degradation of the mutant mRNA through the nonsense-mediated mRNA decay pathway. These data strongly suggest loss of function as the relevant patho-genetic mechanism. PMID- 10807273 TI - Surgical critical care: still at risk? AB - It has been a great honor and privilege to serve as your president and to give this year's presidential address. Considering that this is the last Western Surgical Association presidential address to be given in the 1900s, it seems necessary to mention the rapidly approaching third millennium. With only a little more than 46 days left, I am not particularly excited about having to write a "00" whenever I date something. It just does not seem like a real date to me. I have, however, resisted the temptation to speculate on the 21st century, let alone the next millennium, largely because my crystal ball is no better than any of yours. When I began as a medical student in the mid 1950s, my wildest dreams could not have predicted what dramatic advances would occur in the last half of this century. At that time, the first cardiopulmonary bypass cases were being performed for valvular and congenital disease. Kidneys had only recently been transplanted in Paris, France, and Boston, Massachusetts. Plastic surgery had yet to do a free flap and video-guided endoscopic surgery was still decades away. Intensive care units (ICUs) were just opening up, but they were little more than places where frequent vital signs and reliable intake and outputs were obtained. PMID- 10807274 TI - Pancreatoduodenectomy for chronic pancreatitis: long-term results in 105 patients. AB - HYPOTHESIS: For patients with head-dominant, small-duct chronic pancreatitis who require operative intervention, pancreatoduodenectomy can be performed safely and affords satisfactory pain relief in most. DESIGN: Retrospective case series. Follow-up was complete in 86% of study subjects (average, 6.6 years). SETTING: Tertiary care center. PATIENTS: Among 484 consecutive cases of chronic pancreatitis treated surgically from January 1976 through April 1997, 105 (22%) in which pancreatoduodenectomy was performed were reviewed with regard to criteria for selection, operative procedure, postoperative course. and long-term outcome. MAIN OUTCOME MEASURES: The main outcome measure was degree of pain relief. Additionally, late mortality, cause of death, the presence of endocrine and exocrine insufficiency, and quality of life were recorded. RESULTS: There were 72 men (69%) and 33 women (31%) with a mean age of 51 years (range, 24-77 years). The cause of chronic pancreatitis was alcohol related in 58 patients (55%) and idiopathic in 41 (39%). Clinical manifestations included abdominal pain in 86 patients (82%), obstructive jaundice in 27 (26%), and vomiting in 11 (11%). Suspicion of malignant neoplasm was a concern in 67 patients (64%). Operative morbidity was 32%, and mortality, 3%. Mean hospital stay was 16 days (range, 12 82 days). Survival was significantly lower than that of age-matched controls. Among 66 patients with preoperative pain, pain relief was achieved in 59 (89%); it was complete in 44 patients (67%) and partial in 15 (23%). Operation resulted in a significant increase in patients with normal functional status (73 patients [81%] vs 51 [49%]; P<.001). Forty patients (48%) had diabetes. Steatorrhea was observed in 39 patients (43%), while weight maintenance or gain occurred in 59 (66%). CONCLUSIONS: Pancreatoduodenectomy achieves pain relief and good quality of life in a large percentage of selected patients with small-duct, head-dominant disease and is especially useful when a malignant neoplasm must be excluded. Morbidity and mortality are acceptable in experienced hands. Onset of diabetes and steatorrhea, while reflecting the natural course of the disease, is likely accelerated by pancreatoduodenectomy. PMID- 10807275 TI - Intraoperative carotid evaluation. AB - HYPOTHESIS: Intraoperative duplex scanning can identify technical defects and increase the quality of carotid artery repair. DESIGN: We evaluated 100 consecutive carotid operations in 96 patients (60 men and 36 women) from 1995 to 1998. Spectral-derived peak systolic flow velocities (PSV) were graded (PSV < 100 cm/s, normal laminar flow; PSV 100-150 cm/s, mild or moderate flow disturbance; PSV > 150 cm/s, severe flow disturbance). Prospective criteria for intraoperative revision included PSV greater than 150 cm/s, spectral broadening, and B-mode imaging of intimal flaps or intraluminal debris. Preoperative, intraoperative, and 6-week follow-up duplex scan results were analyzed. SETTING: All patients were evaluated and treated at a single academic institution. INTERVENTIONS: All procedures were performed with the patient under general endotracheal anesthesia; 86% underwent shunting and 70% underwent patching. MAIN OUTCOME MEASURE: Number and type of revisions, patency of repair, residual and recurrent stenosis, and ipsilateral neurologic events. RESULTS: There were 33 intraoperative duplex studies with abnormal findings. Seven involved the common carotid artery and resulted in intraoperative revision of 5 intimal flaps at the site of the proximal clamp. In 11 patients, incomplete eversion endarterectomy resulted in elevated distal intimal flaps in the external carotid artery that were repaired through a separate arteriotomy. There were 15 abnormalities in the internal carotid artery prompting 5 revisions. Five studies with PSV of 100 to 150 cm/s had no defects on B-mode imaging and were observed without treatment. Five false positive studies were attributed to increased flow velocity due to contralateral occlusive discase. At 6 weeks' follow-up, 4 of 5 repaired common carotid arteries were normal on duplex scan and 1 had a mild residual stenosis. Ten of the 11 external carotid repairs were patent and 1 was occluded. Four of the 5 internal carotid artery repairs were normal on postoperative evaluation and 1 had a mild residual stenosis. Of the 10 abnormal internal carotid arteries that were observed, 9 were normal on postoperative duplex and 1 had a mild residual stenosis. One perioperative stroke occurred in a patient with a normal, patent carotid repair. CONCLUSIONS: Intraoperative duplex evaluation of carotid reconstruction is an efficient, sensitive tool that can detect technical lesions that will jeopardize surgical reconstruction. Interpretive judgment is required because all flow disturbances do not dictate surgical intervention. This technique enables the surgeon to maximize the quality of the arterial reconstruction during carotid artery surgery. PMID- 10807276 TI - Initial presentation with stage IV colorectal cancer: how aggressive should we be? AB - HYPOTHESIS: The appropriate surgical treatment of patients with colorectal cancer who are found on initial presentation to have stage IV disease is controversial. With presumed limited life expectancy, the role of primary colon or rectal resection has been questioned, as has the utility of synchronous hepatic resection. DESIGN: A retrospective chart review. SETTING: The University of Chicago Hospitals, Chicago, Ill, a tertiary-care referral center. PATIENTS: One hundred twenty patients were identified through The University of Chicago Hospitals Tumor Registry whose initial presentation showed stage IV colorectal cancer and who underwent laparotomy. MAIN OUTCOME MEASURES: The primary end points of the study were perioperative morbidity and mortality and overall survival. RESULTS: Median survival and 5-year survival were 14.4 months and 10%, respectively. Survival was greater for patients younger than 65 years than for those who were aged 65 years or older (18.3 vs 9.8 months; P = .007). Carcinomatosis was associated with significantly decreased survival when compared with less extensive stage IV disease (6.7 vs 18.1 months; P<.001). Patients who underwent any form of resection of hepatic metastases achieved a survival advantage over those with unresectable liver lesions (median survival, 29.6 vs 10.2 months; P<.001). Overall, 27 patients (22.5%) developed postoperative complications. Seven patients (5.8%) died during the postoperative period. CONCLUSIONS: Age of 65 years or older, carcinomatosis, and extensive (bilobar) liver involvement are associated with decreased survival and increased postoperative morbidity and mortality and may negate any potential benefit patients derive from resection of the primary lesion. A substantial number of patients with synchronous hepatic metastases have protracted survival that justifies resection of the primary colorectal tumor at initial presentation. Despite the presence of stage IV disease, resection of the primary tumor and, when feasible, liver metastases is indicated. PMID- 10807277 TI - Postcholecystectomy abdominal bile collections. AB - HYPOTHESIS: The clinical syndromes caused by bile collections in the abdomen span a wide spectrum and their natural history and risks are not fully appreciated. DESIGN: Analysis of 179 patients with bile fistulas after cholecystectomy, of which 154 patients had undrained bile collections. OBJECTIVE: To characterize the manifestations and natural history of abdominal bile collections. SETTING: A tertiary care teaching hospital. PATIENTS AND METHODS: The clinical findings in 179 patients with bile fistulas resulting from iatrogenic laparoscopic bile duct injuries and other miscellaneous operations between 1990 and 1999 were analyzed. The group of main interest consisted of 154 patients with undrained bile collections. Of these 154 patients, 21% had serious complications, including sepsis and multiorgan failure. The data were analyzed to identify the variables associated with this undesirable outcome. MAIN OUTCOME MEASURES: Symptoms, physical findings, course of illness, and laboratory and imaging findings. RESULTS: The clinical manifestations of intra-abdominal bile collections were initially discounted in 77% of patients, so the problem went unsuspected for a variable and often lengthy period. Abdominal pain and tenderness (bile peritonitis) gradually developed in 18% of patients with bile ascites. There were no differences in the initial clinical findings in this group compared with those who did not develop peritonitis. Nineteen percent of patients with undrained bile collections experienced serious morbidity. The initial clinical findings did not differ in these patients compared with those with a less complicated illness. Serious illness, however, was associated with the following: (1) a longer period of undrained bile (15.4 vs 9.2 days, P=.04) and (2) a higher incidence of infected bile (45% vs 7%, P=.001). CONCLUSIONS: (1) Prominent abdominal pain and tenderness developed in only 21% of patients with abdominal bile collections; (2) the symptoms caused by bile collections were often subtle and their significance was overlooked, which resulted in a delay in diagnosis; (3) the early clinical findings could not distinguish patients who did become critically ill from those who did not; and (4) seriously ill patients more often had delayed drainage and infected bile. Still, failure to drain a bile collection within just 5 days resulted in serious illness in a few patients. Surgeons must watch for the clinical manifestations of bile ascites after laparoscopic cholecystectomy. This diagnosis should be suspected whenever persistent bloating and anorexia last for more than a few days; failure to recover as smoothly as expected is the most common early symptom of bile ascites. If bile collections were promptly diagnosed and drained, the rate of serious illness resulting from this complication would decline. PMID- 10807278 TI - Gastroesophageal disease and nausea: does fundoplication help or hurt? AB - HYPOTHESIS: Nausea associated with gastroesophageal reflux disease is cured by laparoscopic Nissen fundoplication (LNF). DESIGN: Prospective cohort study of unselected patients who underwent LNF from January 1, 1995, through March 31, 1999. Patients were followed up by a physician for 6 to 36 months. SETTING: A large community teaching hospital. PATIENTS: One hundred consecutive patients with gastroesophageal reflux disease who underwent LNF; all patients were followed up. Patients were grouped according to the presence (group A, n = 33) or absence (group B, n = 67) of preoperative nausea. Interventions were LNF, esophageal manometry, 24-hour pH monitoring, and nuclear gastric emptying studies. MAIN OUTCOME MEASURES: Resolution of symptoms after LNF. RESULTS: Nausea was the most common atypical symptom of gastroesophageal reflux disease, occurring in 33 patients (33%). There were no differences in esophageal manometry or 24-hour pH results between groups. There was a female preponderance in group A (55% vs 33%; P = .003). Patients in group A had a higher prevalence of preoperative dysphagia (P = .02). Patients with persistent postoperative nausea had a higher prevalence of cough (P = .003) and dysphagia (P = .009). The LNF was more effective in reducing heartburn (95% reduction) and regurgitation (95% reduction) than cough and dysphagia (60% reduction). There was a 79% reduction in the number of patients with nausea (33 to 7; P<.001). CONCLUSION: Laparoscopic Nissen fundoplication is effective in eliminating nausea associated with gastroesophageal reflux disease and is not contraindicated in these patients. PMID- 10807279 TI - Relative contributions of technetium Tc 99m sestamibi scintigraphy, intraoperative gamma probe detection, and the rapid parathyroid hormone assay to the surgical management of hyperparathyroidism. AB - HYPOTHESIS: Technetium Tc 99m sestamibi scintigraphy, intraoperative gamma probe detection, and the rapid parathyroid hormone assay have been used to permit a directed operation in patients with hyperparathyroidism. We hypothesized that the coordinated use of these techniques might be particularly useful in patients who require a second operation for hyperparathyroidism. DESIGN: Retrospective analysis was performed to determine the specific contribution of these technologies to the surgical management of patients with hyperparathyroidism who underwent evaluation by at least 2 of these techniques between April 1996 and October 1999. SETTING: Patients were evaluated and treated by an endocrine tumor surgery group within a tertiary care referral center. PATIENTS: Coordinated application of 99mTc-sestamibi scintigraphy, intraoperative gamma probe detection, and/or the rapid parathyroid hormone assay was performed in 32 patients. RESULTS: Twenty-eight of 32 patients had primary hyperparathyroidism, 3 had multiple endocrine neoplasia type 1, and 1 had secondary hyperparathyroidism. The surgical procedure was an initial cervical exploration in 19 and a second operative procedure in 13. Parathyroidectomy was successful in all patients. A directed anatomic operation was performed in 24 patients, including 11 patients who underwent second operative procedures and 9 patients who underwent minimally invasive procedures under local anesthesia. A directed operation was facilitated by sestamibi scan in 22 of 24 patients, intraoperative gamma probe detection in 5 of 23 patients, and the rapid parathyroid hormone assay in 15 of 15 patients. CONCLUSIONS: Coordinated application of 99mTc-sestamibi scintigraphy, intraoperative gamma probe detection, and the rapid parathyroid hormone assay allows for successful directed reoperative parathyroidectomy; a minimally invasive procedure may be performed in selected patients. PMID- 10807280 TI - Operative management of diverticular emergencies: strategies and outcomes. AB - HYPOTHESIS: A selective surgical approach using either a 1- or a 2-stage resection is relatively safe and effective in the management of acute complicated colonic diverticulosis. DESIGN: A consecutive cohort study. SETTING: A university hospital. PATIENTS: Eighty-nine consecutive patients who underwent emergency operations for diverticular disease between July 1, 1984, and June 30, 1999. There were 53 male and 36 female patients (mean age, 47 years). The ethnic background was predominantly Mexican American (58 patients [65.2%]). INTERVENTIONS: Resections of the affected colon (n = 83) plus construction of a Hartmann pouch or mucous fistula (n = 72) or primary anastomosis (n = 11). MAIN OUTCOME MEASURES: Morbidity, mortality, and length of hospital stay. RESULTS: Sixty-eight operations were performed for perforation at an annual rate that has increased greater than 75% in the past 15 years. Another 14 patients underwent operations for obstruction, and 7 underwent operations to control unremitting hemorrhage. Surgical therapy included resection of the affected segment of the bowel in 83 (93%) of the 89 patients, and a Hartmann pouch or mucous fistula was added in 72 (81%). A primary anastomosis was performed in 4 (80%) of 5 right sided lesions but in only 7 (8%) of 84 left-sided lesions. Morbidity occurred in 38 (43%) of the 89 patients, and the mortality was 4%, with 4 deaths occurring secondary to sepsis in high-risk patients with perforations (n = 3) or obstructions (n = 1). The average length of hospital stay was 19.7 days (range, 5 80 days). CONCLUSIONS: Emergency operations for diverticular disease are uncommon but may be associated with substantial morbidity and occasional mortality. Complicated diverticulosis may present at a relatively young age, and perforated forms appear to be increasing rapidly in prevalence. Most diverticular lesions can be satisfactorily managed using a selective approach based on resection with either a primary anastomosis or a temporary colostomy. PMID- 10807281 TI - Twenty cases of peristomal pyoderma gangrenosum: diagnostic implications and management. AB - HYPOTHESIS: Our experience with peristomal ulcers suggested that peristomal pyoderma gangrenosum (PPG) is an infrequent and usually unrecognized complication of inflammatory bowel disease. We hypothesized that a review of our experience with PPG would clarify the essentials of its diagnosis, evaluation, and treatment. DESIGN: A case series of 20 consecutive patients with PPG complicating inflammatory bowel disease were treated at our institution between 1986 and 1999. There were 15 women and 5 men. At the time of development of peristomal pyoderma, 10 of 20 patients had a diagnosis of Crohn disease (CD), while 9 had a diagnosis of ulcerative colitis (UC). One patient was diagnosed as having CD only after first developing PPG. MAIN OUTCOME MEASURE: Healing of PPG. INTERVENTIONS: All patients had failed local enterostomal care prior to referral. Debridements and/or stomal revisions were uniformly unsuccessful. Biopsies, when performed, did not provide clinically important information. Treatment was directed toward inflammatory bowel disease, with variable clinical responses to corticosteroids, metronidazole, cyclosporine, sulfasalazine, and infliximab. RESULTS: Ultimately, 13 patients had a diagnosis of CD. Of these patients, 12 (92%) of 13 developed PPG coincident with recurrent disease. Two patients had a remote history of proctocolectomy for UC and subsequent evaluation revealed CD. One patient developed PPG adjacent to a urinary Kock pouch after cystectomy; ultimately, a diagnosis of CD was made. No patients were lost to follow-up, but in 1 case of UC, no evaluation for latent CD was carried out. The final diagnosis was CD disease in 13 (65%) of 20 and UC in 7 (35%) of 20 patients. All PPG ulcers healed completely, within an average of 11.4 months (median, 8 months; range, 1-41 months). Ulcer resolution was achieved with medical therapy alone in 14 (70%) of 20 cases. Resection of active gastrointestinal CD resulted in healing in 5 (83%) of 6 cases. One case healed 2 months after conservative therapy only. CONCLUSIONS: This review of the largest reported series of PPG suggests the following: (1) PPG complicating inflammatory bowel disease is uncommon and often misdiagnosed by clinicians; (2) local wound care measures have little role in the healing of PPG; (3) PPG usually heralds active CD; (4) in patients with prior history of UC, PPG indicates CD until proven otherwise; (5) prolonged medical therapy (11 months), usually with immunosupression, is required for healing of PPG; and (6) if feasible, surgical resection of all active CD leads to the healing of PPG ulcers. PMID- 10807282 TI - Changing patterns of resident operative experience from 1990 to 1997. AB - HYPOTHESIS: Resident operative experience has increased or decreased with respect to 12 specific operations. DESIGN: A retrospective analysis of resident operative experience reported to the Accreditation Committee for Graduate Medical Education for academic years 1990-1997. SUBJECTS: Residents completing an Accreditation Committee for Graduate Medical Education surgical program. MAIN OUTCOME MEASURES: The total number of residents, average number of operations performed per resident, and the most common operations performed. RESULTS: The number of house staff completing surgical residency training programs has remained constant, while operative volume has increased from 1991 to 1997. Comparison of the frequencies of 12 selected operative procedures performed in academic years 1990 1991 and 1996-1997 found increases in the following procedures: carotid endarterectomy (137%), pancreaticoduodenectomy (66.7%), laparoscopic cholecystectomy (64.8%), parathyroidectomy (51.2%), thyroidectomy (19.2%), colectomy (14.1% to 44.4% depending on subtype), and elective infrarenal aortic aneurysm repair (10.7%). Conversely, frequencies decreased for open cholecystectomy (63.4%), open parietal cell vagotomy (40%), modified radical mastectomy (15.2%), gastroesophageal antireflux procedure (10.4%), and subtotal gastric resection (8.93%). Resident experience was essentially unchanged for emergent infrarenal aortic aneurysm repair and laparoscopic proximal gastric vagotomy. CONCLUSIONS: The number and variety of operative interventions in surgical therapeutics is changing. Continued analysis of the operative experience during surgical training will indicate the need for changing requirements for surgical resident experience. The causes of these shifts are not specifically addressed by this study. Perhaps technological advances in the diagnosis and management of surgical patients or the increase in subspecialty training programs have affected the experience of general surgery trainees. PMID- 10807283 TI - Chemoreflexes: an experimental study. AB - HYPOTHESIS: Transmyocardial laser revascularization (TMLR) will not denervate the heart, because it does not destroy all of the afferents. This study was designed to determine if stimulation of cardiac sympathetic and vagal afferents from an area of the left ventricle treated with TMLR could evoke reflex effects, and thus whether TMLR would denervate the heart. METHODS: The effect of TMLR on reflexes evoked by chemically stimulating cardiac afferents was examined in 9 dogs. Bradykinin and capsaicin were applied topically or injected into the left anterior descending coronary artery before and after TMLR and after bilateral vagotomy and sympathectomy. Aortic (AoP) and left ventricular pressures (LVP) and electrocardiography were monitored. The first derivatives of LVP (dP/dt) were calculated. RESULTS: Topical bradykinin elicited variable hemodynamic responses. Topical capsaicin evoked pressor responses, increasing mean (+/- SEM) AoP (105+/ 9 to 115+/-9 mm Hg; P<.001) and positive dP/dt (+dP/dt) (1032+/-81 to 1159+/-10 mm Hg/s; P<.01) before TMLR. Intracoronary capsaicin evoked a depressor response before TMLR. Pressor responses remained intact after TMLR with increases in mean AoP and +dP/dt (115+/-6 to 128+/-5 mm Hg and 1039+/-98 to 1136+/-100 mm Hg/s, respectively; P<.01). Depressor responses also remained intact after TMLR (91+/ 10 vs 101+/-11 mm Hg [P<.02], and 865+/-104 vs 931+/-104 mm Hg/s [P<.05], respectively). Hemodynamic responses were diminished after bilateral vagotomy and abolished after bilateral sympathectomy. CONCLUSION: Since activation of cardiac afferent nerves and reflex responses remained intact after TMLR, but changed after vagotomy or sympathectomy, TMLR does not denervate the heart sufficiently to be the cause of improved angina after TMLR. PMID- 10807284 TI - Use of a microsatellite marker in predicting dysplasia in ulcerative colitis. AB - BACKGROUND: Patients with ulcerative colitis (UC) have an increased risk of developing colorectal cancer. The current screening protocol involves an annual colonoscopy and biopsy after the patient has had the disease for 8 years. This, however, does not prevent the development of colorectal cancer. HYPOTHESIS: A microsatellite marker for IBD1 may identify individuals who are at greater risk of developing dysplasia and therefore colorectal cancer. DESIGN: Case-control study. SETTING: Single surgical practice. PATIENTS AND METHODS: DNA was extracted from peripheral leukocytes of 152 patients: 22 with UC and dysplasia; 48 with UC and no dysplasia; 24 with colorectal cancer; and 58 with noninflammatory bowel disease, nonmalignant gastrointestinal tract disease who were used as control patients. A microsatellite marker for IBD1 (D16S541) was amplified by polymerase chain reaction. Genotypes were identified using autoradiography. RESULTS: Six alleles and 15 genotypes were identified for marker D 16S541. Genotype CC was found in 33% (8/24) of cancer patients but only 12% (7/58) of controls (chi2 = 5.5; P = .02). Thirty-two percent (7/22) of patients with dysplastic UC also had this genotype, whereas only 8% (4/ 48) of patients with nondysplastic UC had the genotype (chi2 = 4.6; P = .03; vs controls: chi2 = 3.1; P = .08). CONCLUSIONS: This microsatellite marker for IBD1, when combined with other markers, has the potential to be used as a screening tool for colorectal cancer and dysplasia in patients with UC. Such a marker would be of particular use in improving the sensitivity and specificity of the current screening protocol for dysplasia and colorectal cancer for patients with UC. PMID- 10807285 TI - Microvessels that predict axillary lymph node metastases in patients with breast cancer. AB - HYPOTHESIS: The density of vasoactive endothelial growth factor receptor 3 immunostained microvessels in primary breast cancers correlates with the incidence of axillary lymph node metastasis. DESIGN: Breast cancer microvessel clusters ("hot spots") were sequentially immunostained for factor VIII, type IV collagen, and vasoactive endothelial growth factor receptor 3. Microvessels were counted under light microscopy at a magnification of x 200. Axillary lymph nodes were evaluated for metastases by light microscopy. SETTING: A multidisciplinary breast cancer clinic and laboratory. PATIENTS: Sixty patients with T2 breast cancers treated by lumpectomy (or mastectomy) and axillary lymphadenectomy. MAIN OUTCOME MEASURES: Putative lymphatic microvessel density compared with axillary metastases. RESULTS: There were 16% (SE, 1.4%) vs 4% (SE, 0.8%) vasoactive endothelial growth factor receptor 3-immunostained microvessels (P<.001), 38% (SE, 3.9%) vs 65% (SE, 3.1%) type IV collagen-immunostained microvessels (P<.001), and 46% (SE, 4.1%) vs 31% (SE, 3.2%) unstained microvessels (P = .004) in node-positive vs node-negative patients, respectively. A fitted logistic model based on the relative percentage of putative lymphatic microvessels to blood microvessels correctly predicted that 23 (96%) of 24 patients would have a low risk and that 26 (96%) of 27 patients would have a high risk of lymph node metastases. Six (67%) of 9 patients predicted to have an intermediate risk had lymph node metastases. CONCLUSION: The odds of a patient with breast cancer having axillary lymph node metastasis increased substantially as the proportion of putative lymphatic microvessels increased and the relative proportion of blood microvessels in angiogenic hot spots decreased (log likelihood = 14.6; chi2 = 53.4; P<.001; area under the receiver operation characteristic curve = 0.97). PMID- 10807286 TI - Reconstruction of small and fragile bile ducts without mucosa-to-mucosa anastomosis. AB - We describe a simple and easy technique for performing choledochojejunostomy without the need to suture the full thickness of the ductal and intestinal walls for patients in whom standard choledochojejunostomy is difficult because the stumps of the residual bile ducts are small and fragile. This technique is useful in partial liver transplantation or after hepatectomy that includes removal of the extrahepatic bile ducts. The procedure involves the placing of external biliary drainage tubes through a Roux-en-Y jejunal loop, positioned transanastomotically, and the use of an external jejunostomy to decompress the loop. The tubes are fixed to the jejunal loop by a purse-string suture and to the duct by simple ligation or a purse-string suture. Anastomosis is performed by suturing the connective tissue and liver parenchyma around the ductal stump to the seromuscular layer of the intestine. Choledochojejunostomy according to this method was performed in 5 cases; the biliary drainage tubes were removed 1 to 4 months after surgery. The only complications were cerebellar infarction and cholangitis, both of which resolved with conservative treatment. We consider that this technique will be helpful as a last-ditch measure when standard choledochojejunostomy, with suturing of the full thickness of the walls of the duct and intestine to secure mucosa-to-mucosa apposition, is impossible because of small and fragile bile ducts. PMID- 10807287 TI - Surgery in New Zealand. AB - Surgery in New Zealand is performed by more than 500 surgeons who serve a population of 3.8 million people. Most of the surgeons are trained in New Zealand under the auspices of the Royal Australasian College of Surgeons. Surgical services are consistent with the highest standards of Western countries. PMID- 10807288 TI - The learning curve for sentinel lymph node biopsy in breast cancer. PMID- 10807289 TI - A Civil War miracle. PMID- 10807290 TI - Identification of domestic violence in the community pediatric setting: need to protect mothers and children. PMID- 10807291 TI - Promoting early detection of human immunodeficiency virus infection among adolescents. AB - While a significant and increasing number of adolescents are infected with the human immununodeficiency virus (HIV), few youth are identified as seropositive and even fewer are linked to medical care and social services. If more youth were identified, transmission to sexual partners and offspring would be reduced and individuals could benefit from treatment. Prior to initiating wide-scale early detection for HIV, we must (1) examine alternative strategies of conducting pretest and posttest counseling; (2) address barriers to prevention and testing within the HIV system of care; and (3) mount community-level intervention campaigns that address youth at high risk of infection. PMID- 10807292 TI - Child and adolescent injury research in 1998: a summary of abstracts submitted to the Ambulatory Pediatrics Association and the American Public Health Association. AB - OBJECTIVE: To describe current research in child and adolescent injury prevention by pediatric and public health investigators for comparison with national recommendations and agendas. DATA SOURCES: Abstracts submitted to the 1998 annual meetings of the Pediatric Academic Societies/Ambulatory Pediatrics Association and the American Public Health Association on injury or violence in children or adolescents. STUDY SELECTION: All abstracts of projects that addressed primarily injury or violence prevention involving children or adolescents. DATA EXTRACTION: For 123 abstracts, 2 coauthors extracted and classified age of the population, type of injury, study design, sizes of the sample and denominator, and type of outcome. RESULTS: Adolescents were the most frequent (49%) age group included. The investigations were concerned most with injuries caused by violence (33%), followed by motor vehicle trauma (14%) and burns (7%). Descriptive surveillance (38%), surveys (32%), and case series (13%) comprised the overwhelming majority of methods used. The studies primarily sought to identify risk factors for injury (32%), describe the victims (20%), or measure knowledge and/or practice (26%). Nine studies (7%) sought to measure the effect of interventions in some way, and only 2 focused primarily on methodology development. CONCLUSIONS: Injury prevention research projects presented at the 1998 Pediatric Academic Societies and American Public Health Association meetings were proportionate to the frequencies of injury by age and by external cause in the United States. However, in comparison with recommendations for agendas of national injury prevention research, more research is needed to improve injury prevention methods and to evaluate interventions. PMID- 10807293 TI - How well does the questionnaire for identifying children with chronic conditions identify individual children who have chronic conditions? AB - BACKGROUND: The Questionnaire for Identifying Children With Chronic Conditions (QuICCC) is an instrument based on a conceptual noncategorical definition that uses parental responses to identify children with chronic conditions for epidemiological purposes. OBJECTIVES: To determine whether the QuICCC is sufficiently valid, sensitive, and specific to be used to identify individual children as having a chronic condition or disability; whether parents are accurate enough that their answers to QuICCC items can be accepted as valid; and what kinds of errors in classification occur when the QuICCC is used to identify children with chronic conditions. METHODS: The sample consisted of 424 children who were patients of 9 physicians in separate practice settings throughout New England. Each physician was briefly trained in the conceptual definition on which the QuICCC is based and then was asked to identify 25 children in his or her practice who met the definition and 25 children who did not meet the definition. The QuICCC was administered to the parents of these children by blinded interviewers via telephone. The QuICCC classification was compared with physician categorization. Discrepant cases were then followed up by asking physicians and parents to answer the original questions a second time. RESULTS: Complete data were available on 379 (89.4%) of 424 children. There was agreement on 89% (kappa = 0.78). The sensitivity was 94%; specificity, 83%; positive predictive value, 86%; and negative predictive value, 92%. Of the 42 discordant cases, 30 parent reports on the QuICCC qualified the child as having a chronic condition when the physician classified the child as being without such a condition. Fewer (n = 12) discrepancies occurred because physicians identified children with chronic conditions that the QuICCC failed to identify. When the questions were readministered at follow-up, physicians corrected errors in rating in 9 cases; mothers changed their answers in 5 instances. In 13 instances the issues were known to both parties and appeared to arise in the "gray zone" or boundary area, where there was disagreement over whether a particular child qualified using the theoretical definition. For 11 children identified as having a chronic condition only by the parent's responses to the QuICCC, physician report appeared to be inaccurate primarily due to the physician's lack of information. In 3 cases where the physician reported the child to have a chronic condition, but the parent did not, the physician appeared to be correct. Follow-up data were incomplete on 1 child. CONCLUSIONS: These data support the validity of parent-generated information for the evaluation of health status. Although these findings should be replicated, this study suggests that the QuICCC may be applicable also as a screening tool for individual child identification, provided that several sources of error are considered. PMID- 10807294 TI - The differentiation of classic Kawasaki disease, atypical Kawasaki disease, and acute adenoviral infection: use of clinical features and a rapid direct fluorescent antigen test. AB - OBJECTIVE: To compare the clinical and laboratory features of children with Kawasaki disease with those with acute adenoviral infection, which may mimic Kawasaki disease. DESIGN: We retrospectively compared the medical records of children with Kawasaki disease and atypical Kawasaki disease with those of children with acute adenoviral infection. All children included were initially evaluated because their primary care physicians were concerned that they might have Kawasaki disease. The utility of a rapid direct fluorescent antigen test for adenovirus was evaluated. Thirty-six children with Kawasaki disease (23 with classic and 13 with atypical presentations) and 7 patients with acute adenoviral infection were studied. SETTING: A tertiary care pediatric hospital. RESULTS: Children with Kawasaki disease were more likely to have conjunctivitis (36 of 36 vs 4 of 7), strawberry) tongues (23 of 36 vs 1 of 7), perineal peeling (19 of 36 vs 0 of 7), and distal extremity changes (22 of 36 vs 0 of 7) than those with acute adenoviral infection. Children with acute adenoviral infection were more likely to have purulent conjunctivitis (3 of 7 vs 1 of 36) and exudative pharyngitis (3 of 7 vs 1 of 35). In addition to pyuria (13 of 26 vs 0 of 6), patients with Kawasaki disease had higher mean white blood cell counts (15.3 +/- 3.5 vs 11.5 +/- 6.0 x 10(9)/L), erythrocyte sedimentation rates (56 vs 42 mm/h), platelet counts (426 vs 259 x 10(9)/L), and levels of alanine aminotransferase (101 vs 18 U/L) than those with acute adenoviral infection. Children with Kawasaki disease had lower mean albumin levels (32 vs 36 g/L). A rapid antigen test for adenovirus had a specificity and sensitivity of 100% compared with viral culture. CONCLUSIONS: Kawasaki disease and acute adenoviral infection can present with many of the same clinical characteristics. A rapid direct fluorescent antigen assay for adenovirus may be a helpful adjunctive test for distinguishing acute adenoviral infection from Kawasaki disease. PMID- 10807295 TI - Identification of violence in the home: pediatric and parental reports. AB - OBJECTIVES: To compare the rates of domestic violence reported by mothers with those identified by physicians, to compare the rates of harsh discipline practices reported by mothers with the rates of abuse identified by physicians, and to examine the relationship between reported domestic violence and harsh discipline practices. DESIGNS: Interviews with parents and pediatricians to compare pediatric detection of domestic violence and child abuse with parental reports of domestic violence and harsh discipline practices. SETTING: Community based pediatric practices in the 13-town greater New Haven, Conn, area. PARTICIPANTS: Of the 23 practices invited, 19 agreed to participate. Of the 2,006 parents of eligible 4- to 8-year-olds asked to participate, 1,886 (94%) completed the Child Behavior' Checklist. Of those invited into the interview portion, 1,148 (83%) completed the 90-minute in-person interview. MAIN OUTCOME MEASURES: Percentages of cases of domestic violence identified by pediatricians and reported by mothers. Percentages of cases of child abuse detected by pediatricians and percentages of mothers reporting that they have hit their children and left a mark. RESULTS: Pediatricians detected domestic violence in 0.3% of the cases, but parents reported domestic violence in 4.2% kappa= 0.106 [95% confidence interval, -0.007 to 0.219]). Pediatricians identified physical abuse of children in 0.5% of the cases, while mothers reported hitting their children and leaving a mark in 21.6% (kappa = 0.003 [95% confidence interval, 0.018 to 0.024]). Mothers reporting domestic violence were significantly more likely to report hitting hard enough to leave a mark (relative risk, 1.6 ([95% confidence interval, 1.09-2.38]) compared with those not reporting domestic violence. Physicians identifying domestic violence were not significantly more likely to report child abuse than those not identifying domestic violence. CONCLUSIONS: Parents report more cases of violence than pediatricians detect. Pediatricians should ask parents directly about domestic violence and harsh discipline. PMID- 10807296 TI - The effect of Kawasaki disease on cognition and behavior. AB - OBJECTIVE: To determine whether there are associated long-term deficits in the cognitive, academic, or behavioral outcomes of children with a previous episode of Kawasaki disease. DESIGN: Cohort analytic study. SETTING: A tertiary care pediatric hospital in Ottawa, Ontario. PARTICIPANTS: Thirty-two patients with a past diagnosis of Kawasaki disease. Siblings of the patients with Kawasaki disease were eligible to be controls. MEASURES: A blinded psychometrist (Y.K.) assessed cognition by the appropriate Wechsler Intelligence scale, academic achievement by the Wechsler Individual Achievement Test, and behavior by the Achenbach Child Behavior Checklist. RESULTS: No differences were found in cognitive or academic measures and the mean scores corresponded closely to national norms. Parents rated their children who had Kawasaki disease as having significantly more internalizing (P<.03) and attentional (P<.02) behavior problems than controls; the risk of a clinically significant behavioral score was 3.3 times greater (P<.03; 95% confidence interval, 1.1-9.9) than for sibling controls. CONCLUSIONS: While no effect on cognitive development or academic performance was demonstrated, these results provide preliminary indication of a post-Kawasaki disease deficit in internalizing and attentional behavior. PMID- 10807297 TI - Resource utilization and contaminated blood cultures in children at risk for occult bacteremia. AB - OBJECTIVE: To measure the increases in resource utilization and hospital charges associated with the evaluation of contaminated blood cultures obtained from emergency department patients at risk for occult bacteremia. DESIGN: Retrospective medical record review. SETTING: Emergency department of an urban, university-affiliated pediatric referral center. PATIENTS: Children aged 3 to 36 months with blood cultures positive for bacterial growth obtained between January 1994 and October 1996. MAIN OUTCOME MEASURES: Increased resource utilization related to contaminated blood culture follow-up, including telephone contact, return emergency department visits, additional diagnostic tests and therapies performed at reevaluation, and hospital admissions. Hospital charges for these additional services were tabulated. RESULTS: Of 8,306 children who had blood cultures drawn, 491 (5.9%) had positive findings. Four hundred eighty-five (98.8%) of these were available for review. Two hundred seventy-six (57%) of 485 were excluded from final analysis. Of the remaining 209 patients at risk for occult bacteremia, 85 (41%) had cultures that grew contaminants only. Follow-up of these 85 patients generated 106 telephone calls, 49 return visits to the emergency department, 102 additional diagnostic tests, 18 doses of parenteral antibiotics, and 12 hospital admissions with a combined length of stay of 24 days. This resulted in additional charges of $78,904, or an additional $642 per true pathogen recovered. CONCLUSIONS: Contaminated blood cultures obtained from children at risk for occult bacteremia lead to substantial increases in resource utilization and hospital charges. These untoward effects should be accounted for in formal decision analyses regarding the management of occult bacteremia. PMID- 10807298 TI - Effect of the baby-friendly initiative on infant abandonment in a Russian hospital. AB - OBJECTIVE: To study whether early mother-infant contact with suckling and rooming in reduces the rate of infant abandonment. DESIGN: The infant abandonment rate was studied at a Russian hospital before and after the introduction of early mother-infant contact with suckling and rooming-in. SETTING: Maternity Hospital 11 , a public hospital in St Petersburg, Russia, was chosen as the site of this study because it recently changed its maternity care practices, implementing portions of the United Nations Children's Fund Baby-Friendly Hospital Initiative. INTERVENTIONS: In mid 1992, Maternity Hospital 11 changed its practices in accord with the Baby-Friendly Hospital Initiative, encouraging early contact, suckling, and rooming-in of the mother and infant from birth to the time of discharge from the hospital. PATIENTS: The location of a mother's maternity hospital is related to her residential district. Maternity Hospital 11 serves an urban working-class community, with most mothers receiving prenatal care. All deliveries at this hospital from 1987 to 1998 were studied. MAIN OUTCOME MEASURE: Infant abandonment. RESULTS: The rate of infant abandonment at Maternity Hospital 11 was studied from 1987 to 1998, 6 years before and 6 years after the implemented changes in mother-infant contact. The mean (+/-SD) infant abandonment rate decreased from 50.3 +/- 5.8 per 10,000 births in the first 6 years to 27.8 +/- 8.7 per 10,000 births in the next 6 years following implementation of the Baby Friendly Hospital Initiative. CONCLUSION: Encouraging early mother-infant contact with suckling and rooming-in may provide a simple, low-cost method for reducing infant abandonment. PMID- 10807299 TI - Improving rheumatologists' screening for alcohol use and sexual activity. AB - OBJECTIVES: To design, implement, and assess the impact of an office-based intervention designed to improve rheumatologists' identification of risk behaviors, especially alcohol use and sexual activity, among adolescents and young adults with chronic rheumatologic conditions. DESIGNS: Prospective intervention study. SETTING: Midwestern academic pediatric rheumatology practice. PARTICIPANTS: Ten attending rheumatologists and fellows and 178 patients (mean age, 18.1 years; 67% female; 88% white; 69% with juvenile rheumatoid arthritis) seen in the practice during the baseline and intervention years. MAIN OUTCOME MEASURES: Change in the rate of screening for alcohol use and sexual activity from the baseline to the intervention year, and physician perceptions of the intervention. RESULTS: Screening for alcohol use increased from 4.2% (9/208) at baseline to 31.6% (56/177) after the intervention (P<.001). Of those patients undergoing screening at follow-up, 20 (36%) of 56 patients reported any alcohol use and 11 (20%) reported current alcohol use. Of those reporting current use, 7 (64%) were counseled or referred. Methotrexate use increased the likelihood of alcohol screening (43% [33/76] vs 26% 123/871; P = .02). Screening for sexual activity increased from 12.4% (27/ 218) to 36.2% (64/177) (P<.001) from baseline to follow-up. Of 52 females undergoing screening at follow-up, 31 (60%) were sexually active. Eleven (41%) of 27 sexually active females were not using contraception other than condoms (4 were not asked about contraception); 7 (82%) of these were referred for contraceptive counseling. Seven rheumatologists completed in-depth semistructured interviews after the intervention. All reported time as a main barrier to screening. Other barriers included logistical problems, discomfort with the subject area, ambivalence about whether risk behavior screening is the province of pediatric rheumatologists, and perceived lack of applicability to their patients. CONCLUSIONS: Despite knowledge and concern about the interaction of immunosuppressive therapy and risk behaviors, few rheumatologists adequately screen the behavior of their adolescent and young adult patients. Time constraints, organizational issues, and physician beliefs remain barriers to widespread screening. PMID- 10807300 TI - A cost-effectiveness analysis of newborn hearing screening strategies. AB - CONTEXT: Congenital hearing loss affects between 1 and 3 out of every 1,000 children. Screening of all neonates has been made possible by the development of portable automated devices. Universal screening is a 2-stage screening process using automated transient-evoked otoacoustic emissions, followed when indicated by automated auditory brain response testing. Targeted screening reserves the 2 stage screening process for those infants at risk for congenital hearing loss. OBJECTIVE: To compare the expected costs and benefits of targeted screening with universal screening for the detection of significant bilateral congenital hearing loss. DESIGN: Cost-effectiveness analysis from the health care system perspective. including costs directly related to screening and initial follow-up evaluation. MAIN OUTCOME MEASURES: Number of cases identified, number of false positives, and cost per case. RESULTS: For every 100,000 newborns screened, universal screening detects 86 of 110 cases of congenital hearing loss, at a cost of $11,650 per case identified. Targeted screening identifies 51 of 110 cases, at $3,120 per case identified. Universal screening produces 320 false-positive results, 304 more than targeted screening. Switching to universal screening from targeted screening would cost an additional $23, 930 for each extra case detected. CONCLUSIONS: Universal screening detects more cases of congenital hearing loss, at the expense of both greater cost and more false-positive screening results. Little is known about the negative impact of false-positive screening and about the benefits of early intervention for congenital hearing loss. Those who advocate adoption of universal screening should be aware not only of the direct costs of universal screening, but of the indirect costs and strategies to increase the benefits of screening. PMID- 10807301 TI - Health care providers' experience reporting child abuse in the primary care setting. Pediatric Practice Research Group. AB - OBJECTIVES: To describe (1) primary care providers' experiences identifying and reporting suspected child abuse to child protective services (CPS) and (2) variables affecting providers' reporting behavior. DESIGN AND METHODS: Health care providers (76 physicians, 8 nurse practitioners, and 1 physician assistant) in a regional practice-based network completed written surveys that collected information about the demographic characteristics of each provider and practice; the provider's career experience with child abuse; and the provider's previous year's experience identifying and reporting suspected child abuse, including experience with CPS. RESULTS: All providers (N = 85) in 17 participating practices completed the survey. In the preceding 1 year, 48 respondents (56%) indicated that they had treated a child they suspected was abused, for an estimated total of 152 abused children. Seven (8%) of 85 providers did not report a total of 7 children with suspected abuse (5% of all suspected cases). A majority of providers (63%; n = 29) believed that children who were reported had not benefited from CPS intervention, and 21 (49%) indicated that their experience with CPS made them less willing to report future cases of suspected abuse. Providers who had some formal education in child abuse after residency were 10 times more likely to report all abuse than were providers who had none. CONCLUSIONS: Primary care providers report most, but not all, cases of suspected child abuse that they identify. Past negative experience with CPS and perceived lack of benefit to the child were common reasons given by providers for not reporting. Education increases the probability that providers will report suspected abuse. PMID- 10807302 TI - Violence prevention in the emergency department: clinician attitudes and limitations. AB - OBJECTIVE: To assess emergency department (ED) clinicians' attitudes and behaviors regarding identification, assessment, and intervention for youth at risk for violence in the ED. DESIGN: Anonymous, cross-sectional written questionnaire. SETTING: The EDs of 3 urban hospitals. SUBJECTS: Emergency medicine residents and faculty, pediatric residents, pediatric emergency medicine fellows and faculty, and ED nurses. RESULTS: A total of 184 (88%) of 208 clinicians completed the questionnaire. Only 15% correctly recognized the lack of existing protocols for addressing youth violence. Clinicians reported being most active in identification of at-risk youth (93% asking context of injury and 82% determining relationships of victim and perpetrator), with pediatricians being more active than general ED clinicians (87% vs 68%; P<.01). Clinicians less often reported performing assessments or referrals of at-risk youth. Nurses and physicians were no different in their reported identification, assessment, or referral behaviors. Barriers identified include concern over upsetting family members, lack of time or skills, and concern for personal safety. Additional clinician training, information about community resources, and specially trained on-site staff were noted by respondents as potential solutions. CONCLUSIONS: Emergency department clinicians recognize the need for evaluation of youth at risk for violence. They are able to identify violently injured youth, but less often perform risk assessment to guide patients to appropriate follow-up resources. Further investigation should address clinician barriers to the complete care of violently injured youth in the ED. PMID- 10807304 TI - Growth and determinants of access in patient e-mail and Internet use. AB - OBJECTIVES: To measure the rate of access to and use of the Internet and e-mail, to determine sociodemographic predictors of access, and to measure the change in Internet and e-mail access over a 1-year interval. DESIGN: Survey study. Comparison of data with those from a similar survey from 1998. SETTING: Emergency department of a large urban pediatric teaching hospital. PARTICIPANTS: Primary caretakers of pediatric patients or the patients themselves if aged 16 years or older. MAIN OUTCOME MEASURES: Use of and access to the Internet or e-mail. RESULTS: We surveyed 214 individuals: 72.8% use or have access to the Internet, e mail, or both, an increase from 52.2% in the 1998 survey (P<.001), and 48.5% regularly use the Internet or e-mail, compared with 43.1% in 1998 (P = .32). Outside the home, access is primarily at work (52.2%), schools (8.9%), public libraries (11.5%), and friends' and relatives' houses (16.7%). Internet use and access are linearly correlated with income (r = 0.43; P<.001). White patients are more likely to have access (odds ratio, 2.6; 95% confidence interval, 1.3-5.4; P<.001) than black or Asian patients, whereas those of Hispanic ethnicity are less likely to have access (odds ratio, 0.20; 95% confidence interval, 0.09-0.43; P<.001). However, after adjustment for race and Hispanic ethnicity, only income was a significant predictor of family access to the Internet and e-mail. CONCLUSIONS: During the past year, many patients have gained access to the Internet and e-mail, although rates of regular use have remained steady. This access is often from outside the home. Furthermore, access is directly related to income and is unevenly distributed across racial and ethnic groups. PMID- 10807303 TI - Coordination of specialty referrals and physician satisfaction with referral care. AB - OBJECTIVES: To describe how physicians coordinate patient care for specialty referrals and to examine the effects of these activities on referring physicians' satisfaction with the specialty care their patients receive and referral completion. DESIGN AND METHODS: Prospective study of a consecutive sample of referrals (N = 963) made from the offices of 122 pediatricians in 85 practices in a national practice-based research network. Data sources included a physician survey completed when the referral was made (response rate, 99%) and a physician survey and medical record review conducted 3 months later (response rate, 85%). Referral completion was defined as receipt of written communication of referral results from the specialist. RESULTS: Pediatricians scheduled appointments with specialists for 39.3% and sent patient information to specialists for 50.8% of referrals. The adjusted odds of referral completion were increased 3-fold for those referrals for which the pediatrician scheduled the appointment and communicated with the specialist compared with those for which neither activity occurred. Referring physicians' satisfaction ratings were significantly increased by any type of specialist feedback and were highest for referrals involving specialist feedback by both telephone and letter. Elements of specialists' letters that significantly increased physician ratings of letter quality included presence of patient history, suggestions for future care, follow-up arrangements, and plans for comanaging care; only the inclusion of plans for comanaging patient care was significantly related to the referring physicians' overall satisfaction. CONCLUSIONS: Better coordination between referring physicians and specialists increases physician satisfaction with specialty care and enhances referral completion. Improvements in the referral process may be achieved through better communication and collaboration between primary care physicians and specialists. PMID- 10807305 TI - Making history: Thomas Francis, Jr, MD, and the 1954 Salk Poliomyelitis Vaccine Field Trial. AB - This article focuses on the poliomyelitis vaccine field trial directed by Thomas Francis,Jr, MD, of the University of Michigan Vaccine Evaluation Center and sponsored by the National Foundation for Infantile Paralysis (NFIP) or, as it was better known to the public, the March of Dimes. It was a landmark in the widescale testing of a vaccine and the ethical use of human subjects. Millions of American parents readily volunteered their healthy children to participate. A total of 150,000 volunteers, including schoolteachers, physicians, nurses, and health officers all endorsed the study and donated their time and effort to make it successful. Avoiding the use of marginalized groups, the field trial purposefully did not involve institutionalized children; instead, it was based in 15,000 public schools in 44 of the 48 states as clinic sites. A group of 650,000 children received some type of injection, either the vaccine or a placebo, and another 1.18 million served as controls. The field trial depended, most essentially, on both public support and the participation of millions of children who remained enrolled in a study that required a series of 3 injections and a 6 month evaluation period. Enlisting the huge number of participants presented practical examples of the difficulties in experimenting on human subjects. On April 26, 1954, Randy Kerr, a participant or "Polio Pioneer" as the children involved were called, received the first inoculation of the Salk poliomyelitis vaccine. The nationwide study "designed to test the safety and efficacy" of the Salk vaccine had officially begun. PMID- 10807306 TI - Radiological case of the month. Pyriform sinus fistula to the left lobe of the thyroid. PMID- 10807307 TI - Picture of the month. Congenital aniridia. PMID- 10807308 TI - Pathological case of the month. Proteus syndrome. PMID- 10807309 TI - Sudden infant death syndrome and co-sleeping. PMID- 10807310 TI - Bed sharing is not a "consumer product". PMID- 10807311 TI - Cellular signaling in critical care--putting the pieces together. PMID- 10807312 TI - Cytokine signaling--regulation of the immune response in normal and critically ill states. AB - Cytokines are produced during the activation of innate and acquired immunity, and are the principal means for intercellular communication of a microbial invasion. Cytokines serve to initiate the inflammatory response and to define the magnitude and the nature of the acquired immune response. The response of critically ill patients to their injury and/or invading pathogens is dependent, in large part, on the pattern of cytokines which are produced. The immunologic response of critically ill patients can vary from a strongly proinflammatory response, characterized by increased production of tumor necrosis factor-alpha, interleukin (IL)-1, interferon (IFN)-gamma, and IL-12 to one predominantly of anergy, characterized by increased production of T(H)2 cytokines, like IL-10 and to IL-4. Therapeutic efforts to modify the host immune response in critical illness will require a more thorough understanding of the cytokine milieu and the factors that determine their production. PMID- 10807313 TI - Chemokine signaling in inflammation. AB - The events that lead to an inflammatory response are characterized by recognition of the site of injury by inflammatory cells, specific recruitment of subpopulations of leukocytes into tissue, removal of the offending agent and "debridement" of the injured cells/tissue, and repair of the site of injury with attempts to reestablish normal parenchymal, stromal, and extracellular matrix relationship. The molecular regulation of this complex physiologic process involves the interaction between cell surface, extracellular matrix, and soluble mediators, such as chemokines. Chemokine activities are mediated through G protein coupled receptors. This is the largest known family of cell-surface receptors, which mediate transmission of stimuli as diverse as hormones, peptides, glycopeptides, and chemokines. In this review, we will focus on the signaling pathways involved in the production and function of chemokines as they relate to the inflammatory response. PMID- 10807314 TI - Lipid mediators in the pathophysiology of critical illness. AB - Inflammatory lipid mediators are produced by the metabolism of membrane phospholipids following a number of different stimuli. These mediators lead to a variety of cellular and systemic responses which contribute to the manifestations of the systemic inflammatory response syndrome in the critically ill patient. These mediators include platelet-activating factor and the eicosanoids, including prostaglandins, thromboxanes, leukotrienes, and HETEs. This review seeks to evaluate the current role of these mediators in the pathophysiology of critical illness. We will focus on recent studies concerning the modulation of these pathways as a potential therapeutic strategy for management of these critically ill patients. This includes the gamut from nutritional strategies to alter the cellular membrane lipid composition, thereby effecting the substrate available to produce these lipid byproducts, to intracellular inhibitors to alter production of these mediators, to receptor blockage and enhanced clearance to inhibit their effects. PMID- 10807316 TI - Biology of heterotrimeric G-protein signaling. AB - The G proteins are components of a complex membrane signaling system designed to modulate extracellular signals as they are transmitted into the cell. The principal components are the receptor, the G proteins including the alpha, beta, gamma subunits and the effector. Associated with these molecules are several molecular processes by which the signal is transmitted, and regulated including desensitization. Molecules such as arrestin, the RGS (regulators of the G-protein signaling) as well as downstream kinases associated with cyclic AMP are key to regulating the G protein signal. Membrane lipids are key for both anchoring this signal system to the plasma membrane but also in defining the signaling process. Through understanding the biology of the signal system, a number of diseases have been linked to dysfunction of the G protein system. It is clear that this important membrane signal system will become the target for more intense investigation and pharmacologic manipulation to treat critical illness. PMID- 10807315 TI - Biology of nitric oxide signaling. AB - The free radical nitric oxide (NO) has emerged in recent years as a fundamental signaling molecule for the maintenance of homeostasis, as well as a potent cytotoxic effector involved in the pathogenesis of a wide range of human diseases. Although this paradoxical fate has generated confusion, separating the biological actions of NO on the basis of its physiologic chemistry provides a conceptual framework which helps to distinguish between the beneficial and toxic consequences of NO, and to envision potential therapeutic strategies for the future. Under normal conditions, NO produced in low concentration acts as a messenger and cytoprotective (antioxidant) factor, via direct interactions with transition metals and other free radicals. Alternatively, when the circumstances allow the formation of substantial amounts of NO and modify the cellular microenvironment (formation of the superoxide radical), the chemistry of NO will turn into indirect effects consecutive to the formation of dinitrogen trioxide and peroxynitrite. These "reactive nitrogen species" will, in turn, mediate both oxidative and nitrosative stresses, which form the basis of the cytotoxicity generally attributed to NO, relevant to the pathophysiology of inflammation, circulatory shock, and ischemia-reperfusion injury. PMID- 10807317 TI - Regulation of macrophage activation and inflammation by spermine: a new chapter in an old story. AB - Spermine, a naturally occurring polyamine, is a ubiquitous structural component of all eukaryotic cells. Regenerating tissues produce higher levels of spermine, and injured or dying cells release spermine into the extracellular milieu, so that tissue levels increase significantly at inflammatory sites of infection or injury. Recent research has focused on delineating the significance of spermine accumulation in the inflammatory process. The discovery that spermine is a negative regulator of macrophage activation provided a mechanism by which spermine influences the biology of inflammation. Mechanistic studies indicate that spermine is incorporated into macrophages and restrains the innate immune response. This anti-inflammatory process is facilitated by the negative acute phase protein, fetuin. PMID- 10807318 TI - MAP kinase pathways activated by stress: the p38 MAPK pathway. AB - A stress-activated serine/threonine protein kinase, p38 mitogen-activated protein kinase (p38 MAPK), belongs to the MAP kinase superfamily. Diverse extracellular stimuli, including ultraviolet light, irradiation, heat shock, high osmotic stress, proinflammatory cytokines and certain mitogens, trigger a stress regulated protein kinase cascade culminating in activation of p38 MAPK through phosphorylation on a TGY motif within the kinase activation loop. p38 MAPK appears to play a major role in apoptosis, cytokine production, transcriptional regulation, and cytoskeletal reorganization, and has been causally implicated in sepsis, ischemic heart disease, arthritis, human immunodeficiency virus infection, and Alzheimer's disease. The availability of specific inhibitors helps to clarify the role that p38 MAPK plays in these processes, and may ultimately offer therapeutic benefit for certain critically ill patients. PMID- 10807319 TI - Hypoxic signal transduction in critical illness. AB - Derangements in tissue perfusion occur during critical illness, and the resulting deficit in oxygen delivery may play an important role in the pathogenesis of hemorrhagic and septic shock. Cells and organisms have developed a variety of adaptive strategies to maintain adequate energy production to maintain normal cellular function under hypoxic conditions. Recent studies from our laboratory suggest that certain proinflammatory cytokines, which are likely to be elaborated during or after shock, can interfere with the ability of cells to adapt to hypoxia, and thereby contribute to the development of organ system dysfunction. PMID- 10807320 TI - Ceramide mediates radiation-induced death of endothelium. AB - The sphingomyelin (SM) pathway is an ubiquitous, evolutionarily conserved signaling system, analogous to conventional systems such as the cAMP and phosphoinositide pathways. Ceramide is generated from SM by the action of a neutral or acid SMase, or by de novo synthesis coordinated through the enzyme ceramide synthase. Once generated, ceramide may serve as a second messenger in signaling responses to physiologic or environmental stimuli, or may be converted to a variety of structural or effector molecules. In the radiation response, ceramide serves as a second messenger in initiating apoptosis, while some of its metabolites block apoptosis. In certain cells, such as endothelial, lymphoid and haematopoietic cells, ceramide mediates apoptosis while in others ceramide may serve only as a co-signal for or play no role in the death response. Regulated ceramide metabolism may determine the balance between pro- and anti-apoptotic signals, and hence, the intensity of the apoptotic response, thus constituting a mechanism of radiation sensitivity or resistance. This paradigm may offer new opportunities for modulation of the radiation effects in the treatment of cancer. Chemical modifiers of ceramide metabolism may be useful to enhance the therapeutic effects or reduce the toxicity of radiation treatment. PMID- 10807321 TI - Actin cytoskeleton and cell signaling. AB - The role of the actin cytoskeleton in the function of eukaryotic cells is ubiquitous. Regulation of actin polymerization allows cells to control their shape, to move, divide, secrete, and phagocytose. Actin filaments provide strength, connections to other cells and the extracellular matrix, paths for intracellular transport and a scaffold for generating force. Recently, a number of signal transduction pathways have been identified that regulate actin polymerization and contractility. GTP-binding proteins, protein kinases, phosphoinositide kinases, and protein phosphatases all play important roles in determining the location and extent of actin polymerization and contractility of actin/myosin filaments. These pathways allow cells to respond to extracellular signals to regulate movement, the tone of vascular smooth muscle cells, secretion, and phagocytosis. Some pathogens use signal transduction pathways that regulate actin polymerization to invade cells. The signal transduction pathways that regulate actin-dependent events are the focus of this review. PMID- 10807322 TI - NF-kappaB activation. AB - Binding sites for the transcriptional regulatory factor nuclear factor kappa B (NF-kappaB) are present in the promoter regions of many of the proinflammatory cytokines and immunoregulatory mediators important in inducing acute inflammatory responses associated with critical illnesses. Because increased activation of NF kappaB leads to enhanced expression of these proinflammatory mediators, NF-kappaB activation may be a central event in the development of multiple organ dysfunction associated with infection, blood loss, and ischemia-reperfusion injury. NF-kappaB is normally retained in the cytoplasm through its association with the inhibitory molecule I kappaB. Phosphorylation, ubiquination, and proteolysis of I kappaB allows NF-kappaB to translocate to the nucleus and induce transcription, once associated with the transcriptional cofactor CBP. The transcriptional activity of NF-kappaB can be regulated at multiple steps, including the amount of I kappaB present, NF-kappaB subunit composition, and competition for CBP binding. Because of the central role that NF-kappaB occupies in modulating immunoregulatory responses, further understanding of its regulation will be important in designing future therapies able to prevent or minimize acute inflammatory injury associated with critical illness. PMID- 10807323 TI - Apoptosis in sepsis. AB - Sepsis demonstrates a marked dysregulation of the immune system in its ability to fight infection. Previous models have focused on the mechanisms which upregulate and sustain the heightened immune response without addressing the role of down regulation effectors. Attention has been drawn to these down-regulating mechanisms and their precise role in the pathophysiology of sepsis. Apoptosis is an evolutionarily conserved, energy-dependent mode of cell death requiring the initiation and regulation of complex genetic programs. It is the body's main method of getting rid of cells which are in excess, damaged, or no longer needed in a controlled manner. The role of this cellular phenomenon in physiology and pathophysiology has been the subject of intense scrutiny over the last decade. Much work has demonstrated that dysregulation of apoptosis does occur in immune and nonimmune cells in in vitro and in vivo models of sepsis. The difficulty has been in tying the phenomenology of apoptosis into the pathophysiology of sepsis. Further work is needed to make these connections to elucidate rational approaches for clinical applications of immunomodulation in sepsis. PMID- 10807324 TI - Nasally inhaled dornase alfa in the postoperative management of chronic sinusitis due to cystic fibrosis. AB - OBJECTIVE: To determine the benefit of nasally inhaled dornase alfa in cystic fibrosis (CF) sinusitis. DESIGN: Retrospective chart review comparing postsurgical course, radiographic studies, and pulmonary function test results in patients who were treated with nasally inhaled dornase alfa with those in patients who were not treated with dornase alfa. PATIENTS: Twenty consecutive patients with CF who underwent functional endoscopic sinus surgery from 1993 to 1997 were included in the study. Treatment with nasally inhaled dornase alfa was initiated in 5 of the 20 patients after they underwent functional endoscopic sinus surgery. SETTING: Tertiary care academic center. RESULTS: The dornase alfa treated patients had less mucosal edema and no polyps at serial endoscopy over 3 years compared with the non-dornase alfa-treated patients. The patients who received nasally inhaled dornase alfa also underwent fewer revision functional endoscopic sinus surgical procedures (1.6 vs 3.2), even though there was essentially no change in pulmonary function test results in these patients. CONCLUSIONS: Sinusitis continues to be a major cause of morbidity in patients with CF. Symptomatic patients frequently require multiple drug regimens, including long-term systemic antibiotic therapy, topical and systemic steroid therapy, and antibiotic nasal irrigations. This preliminary study indicates the potential impact of nasally inhaled dornase alfa in controlling postoperative symptoms in CF sinusitis. PMID- 10807325 TI - Impact of tympanostomy tubes on child quality of life. AB - BACKGROUND: The objective benefits of tympanostomy tubes for otitis media are well established, but the subjective impact of surgery on child quality of life (QOL) has not been systematically studied. OBJECTIVES: To determine the subjective impact of tympanostomy tubes on child QOL, and to compare the variability in QOL before surgery with that observed after surgery. DESIGN: Prospective, observational, before-and-after trial. SETTING: Fourteen referral based pediatric otolaryngology practices in the United States. PATIENTS: Consecutive (64%) and convenience (36%) sample of 248 children (median age, 1.4 years) with otitis media scheduled for bilateral tympanostomy tube placement as an isolated surgical procedure. INTERVENTION: Tympanostomy tubes were inserted as part of routine clinical care. Validated measures of QOL (OM-6 survey), satisfaction with health care decision (Satisfaction With Decision Scale), and satisfaction with office visit; surveys were completed at baseline (visit 1), at surgery (visit 2), and after surgery (visit 3). MAIN OUTCOME MEASURES: Short-term changes in QOL before surgery (visit 1 to visit 2) and after surgery (visit 2 to visit 3). RESULTS: Changes in QOL before surgery were mostly trivial, and were smaller than changes observed after surgery (P<.001). Large, moderate, and small improvements in QOL occurred after surgery in 56%, 15%, and 8% of children, respectively. Physical symptoms, caregiver concerns, emotional distress, and hearing loss were most improved, but significant changes were also seen for activity limitations and speech impairment. Trivial changes occurred in 17% of children, and 4% had poorer QOL. Predictors of poorer QOL were otorrhea 3 or more days (10% of variance) and decreased satisfaction with surgical decision (3% of variance). Hearing status, child age, type of otitis media (recurrent vs chronic), and office visit satisfaction were unrelated to outcome. CONCLUSIONS: Tympanostomy tubes produce large short-term improvements in QOL for most children. The best outcomes occur when postoperative otorrhea is absent or minimal, and when parents are satisfied with their initial decision to have surgery. Further research is needed to document the long-term impact of tubes on child QOL. PMID- 10807326 TI - Anatomical basis of sleep-related breathing abnormalities in children with nasal obstruction. AB - OBJECTIVE: To define, in a group of children with nasal obstruction, the anatomical differences that differentiate those with quiet, unobstructed nocturnal respiration from those with obstructive sleep-related breathing abnormalities (snoring and obstructive sleep apnea). DESIGN: Case series. PATIENTS: Fifty-nine children aged 3 to 13 years (35 boys and 24 girls) with nasal obstruction and without tonsillar hypertrophy, known craniofacial syndromes, or neuromuscular diseases were included in the study. MAIN OUTCOME MEASURES: Each patient was categorized as to severity of nocturnal obstructive breathing symptoms. Angular and linear cephalometric measurements were used for assessment of craniofacial features. Clinical symptom scores were correlated with the cephalometric measurements. RESULTS: Significant craniofacial abnormalities were identified in patients prone to obstructive breathing patterns: increased flexure of the cranial base and bony nasopharynx, opening of the gonial angle, shortened mandibular length, dorsocaudal location of the hyoid, reduced posterior airway space, and increased velar thickness. CONCLUSIONS: A number of anatomical abnormalities may contribute to sleep-related abnormal breathing in otherwise normal children with nasal obstruction. Our results suggest that symptomatic children show some of the same skeletal and soft-tissue configurations that are found in adults with obstructive sleep apnea. While adenoidectomy is generally an effective treatment in children with obstructive sleep-related breathing abnormalities, the underlying craniofacial variances that remain after adenoidectomy may predispose these patients to redevelopment of obstructive breathing abnormalities in adulthood. PMID- 10807327 TI - Radiofrequency tissue volume reduction of the soft palate in simple snoring. AB - BACKGROUND: Snoring is common and often associated with social morbidity. Current therapies are generally unsatisfactory, but radiofrequency tissue volume reduction (RFTVR) palatoplasty offers a new approach. OBJECTIVE: To assess the outcomes and morbidity associated with RFTVR palatoplasty. DESIGN: Open, prospective trial. SETTING: Tertiary referral center. PATIENTS: 20 adults with loud habitual snoring without clinically significant obstructive sleep apnea. INTERVENTIONS: Three treatments with RFTVR to the middle, distal, and proximal thirds of the midline of the soft palate. MAIN OUTCOME MEASURES: Clinical assessment (visual analog scores) before and after each treatment, polysomnography (with sound intensity measurements), and lateral cephalometry performed prior to the first treatment and 2 months following the final treatments. RESULTS: After treatment, there was a significant overall improvement in the snoring visual analog score (7.5+/-1.5 to 4.6+/-2.5; P<.001), a small reduction in the proportion of sleep spent snoring at 50 to 60 dB (P = .03), and mild pain that was controlled with simple analgesia. There were no long-term adverse effects. Individual response could not be predicted by demographic, polysomnographic, or cephalometric data. Treatment of the proximal third of the soft palate was associated with fewer adverse effects but also seemed less effective than at the other sites. CONCLUSIONS: (1) The RFTVR palatoplasty is well tolerated with very low morbidity. (2) It is associated with subjective improvement in snoring in most patients. (3) Placement of lesions seems to influence outcome. (4) The improvement is accompanied by a marginal change in objective measurements, suggesting either an acoustic change independent of sound intensity or a placebo effect. (5) A randomized controlled trial is needed to further evaluate this therapy. PMID- 10807328 TI - Prevention of otitis media with effusion by repeated air inflation in a monkey model. AB - OBJECTIVES: To test the following hypotheses that (1) middle ear (ME) air inflation prevents the development of otitis media with effusion in a monkey model of functional eustachian tube obstruction, and (2) ME inflation treatment of otitis media with effusion can cause artifactual clinical improvements due to fluid displacement from the tympanum to the adjacent airspaces. DESIGN: Randomized controlled trial. SUBJECTS: Twelve cynomolgus monkeys. INTERVENTIONS: Eustachian tube dysfunction was induced by botulinum paralysis of the right tensor veli palatini muscle in all monkeys. Before and on study days 9, 15, and 21 after paralysis, the presence or absence, and distribution of ME effusion were documented using magnetic resonance imaging (MRI). Right and left ears were examined twice daily for 21 days using tympanometry, and right ME air inflation (n = 6 ears) or sham inflation (n = 6 ears) was done immediately after those examinations if the ME pressure was -100 mm H2O or less. On 10 of the scheduled MRI evaluations, the MRI was repeated immediately after an inflation to document the possible redistribution of fluid within the ME caused by the maneuver. RESULTS: Middle ear pressure remained within normal limits for the follow-up period in 11 of the 12 nonparalyzed left ears, in none of the 6 sham-inflated right ears, and in 3 of the 6 air-inflated right ears. Three air-inflated right ears developed flat tympanograms (ie, days 14 through 16). Magnetic resonance imaging documented inflammation and fluid in 1 of the 11 nonparalyzed left ears and in all sham-inflated right ears. Lesser degrees of inflammation and effusion based on MRI evaluations were noted for the 3 air-inflated right ears that retained near-ambient pressures when compared with the right 3 ears that developed a flat tympanogram. The MRI measure of effusion quantity within the tympanum was decreased acutely after inflation, but was simultaneously increased in the adjacent airspaces of the temporal bone. CONCLUSIONS: Repeated air inflation prevented the development of otitis media with effusion in 50% of the ears with functional eustachian tube obstruction. Postinflation MRI documented the displacement of fluid by inflation from the tympanum to the mastoid and petrous air cells. Using standard clinical evaluations such as tympanometry and otoscopy, this fluid redistribution can cause a false diagnosis of improvement. PMID- 10807329 TI - Vibration does not improve results of the canalith repositioning procedure. AB - OBJECTIVE: To determine whether, in patients with benign paroxysmal positional vertigo (BPPV), the canalith repositioning procedure performed with vibration applied over the mastoid bone of the affected ear is more effective in resolving the symptoms and preventing recurrence of BPPV than the procedure performed without vibration. DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Ninety-four patients diagnosed as having BPPV involving the posterior semicircular canal. INTERVENTIONS: Patients were assigned to one of 2 treatment groups: the canalith repositioning procedure with vibration (n=44) and with no vibration (n=50). MAIN OUTCOME MEASURES: Effectiveness of treatment was determined through clinical reevaluation or reported through a telephone interview 1 week after treatment. Intensity of symptoms was quantified on a scale of 1 to 3 (mild, moderate, or severe); effectiveness of treatment was categorized on a scale of 1 to 4 (cure, much better, better, or no change). Rate of recurrence was determined through later clinical reevaluation or a telephone interview. RESULTS: At 1 week, 57 of the 94 patients were cured and 16 were much better, providing a 78% overall success rate. There was no significant difference in effectiveness of the treatment or the frequency of reoccurrence of BPPV between the vibration and no-vibration groups as determined from the Kaplan-Meier product-limit method and log-rank test. Rate of recurrence was 47% at a maximum follow-up of 5.25 years. CONCLUSIONS: Our results suggest that, while the canalith repositioning procedure is effective in the treatment of BPPV, vibration applied during the maneuver does not significantly affect short-term or long-term outcomes. PMID- 10807330 TI - Resistant bacteria in the adenoids: a preliminary report. AB - OBJECTIVE: To determine the incidence of resistant bacteria in adenoid cultures from children with and without middle ear disease and rhinosinusitis symptoms. DESIGN: Children meeting the requirement for tympanostomy tube placement underwent an adjuvant adenoidectomy for symptoms of adenoid hypertrophy or recurrent rhinosinusitis. Adenoid tissue and coexisting middle ear fluid, if present, were cultured. SETTING: Tertiary referral children's hospital with community-based satellite clinics. PATIENTS: Forty-six patients ranging in age from 1 to 11 years (68% <3 years) with recurrent or persistent otitis media and symptoms of adenoid hypertrophy or rhinosinusitis (study patients) underwent tympanostomy tube placement and adenoidectomy with culture of the adenoids and middle ear effusions. Eighteen patients with adenoid hypertrophy without ear disease or rhinosinusitis were used as controls. INTERVENTIONS: Tympanostomy tube placement and adenoidectomy. MAIN OUTCOME MEASURES: Presence or absence of resistant bacteria. RESULTS: Resistant bacteria were found in cultures of the adenoids in 56% (26/46) of the study group compared with 22% (4/18) of the control patients (P<.02). Also, strains of Streptococcus pneumoniae, Haemophilus influenzae, or Moraxella catarrhalis were found in cultures from 78% (36/ 46) of the study group, compared with 44% (8/18) of those from the control group (P<.01). Resistant isolates were found in 65% (23/35) of the S. pneumoniae, 37% (18/49) of the H. influenzae, and 100% (19/19) of the M. catarrhalis cultures from the adenoids or middle ear spaces. CONCLUSION: Resistant bacteria are present in significant amounts in the adenoids of children with middle ear disease and rhinosinusitis symptoms compared with patients without those diseases or symptoms. PMID- 10807331 TI - Clinical characterization of genetic hearing loss caused by a mutation in the POU4F3 transcription factor. AB - OBJECTIVES: To describe the detailed auditory phenotype of DFNA15, genetic hearing loss associated with a mutation in the POU4F3 transcription factor, and to define genotype-phenotype correlations, namely, how specific mutations lead to particular clinical consequences. DESIGN: An analysis of clinical features of hearing-impaired members of an Israeli family, family H, with autosomal dominant inherited hearing loss. SETTING: Department of Human Genetics and Molecular Medicine, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel; Department of Audiology, Rabin Medical Center, Petah Tiqwa, Israel; and audiological centers. PARTICIPANTS: Clinical features of 11 affected and 5 unaffected individuals older than 40 years from family H were studied. Mutation analysis was performed in 6 presymptomatic individuals younger than 30 years; clinical features were analyzed in 4 of these family H members. INTERVENTIONS: Hearing was measured by pure-tone audiometry and speech audiometry on all participating relatives of family H. Immittance testing (tympanometry and acoustic reflexes), auditory brainstem response, and otoacoustic emissions were done in a selected patient population. RESULTS: The patients presented with progressive high-tone sensorineural hearing impairment, which became apparent between ages 18 and 30 years. The hearing impairment became more severe with time, eventually causing significant hearing loss across the spectrum at all frequencies. CONCLUSIONS: Our results indicate that POU4F3 mutation-associated deafness cannot be identified through clinical evaluation, but only through molecular analysis. Intrafamilial variability suggests that other genetic or environmental factors may modify the age at onset and rate of progression. PMID- 10807332 TI - A family with autosomal dominant inherited dysmorphic small auricles, lip pits, and congenital conductive hearing impairment. AB - DESIGN: We examined 3 generations in a family for congenital conductive hearing impairment, dysmorphic small auricles, and lip pits. SETTING: Department of Otorhinolaryngology, University Hospital Nijmegen, Nijmegen, the Netherlands. RESULTS: Seven members of the family had bilateral dysmorphic auricles. Three subjects had either a pit or dimple in the lip. Two subjects had congenital conductive hearing impairment. CONCLUSION: Using gene linkage, we confirmed that these autosomal dominant inherited branchial anomalies present a new separate branchial arch syndrome. PMID- 10807333 TI - Presence of 22q11 deletion in postadenoidectomy velopharyngeal insufficiency. AB - BACKGROUND: Velopharyngeal insufficiency is an uncommon complication of adenoidectomy. Persistent velopharyngeal insufficiency following adenoidectomy (VIA) may occur in association with an unrecognized syndrome, such as velocardiofacial syndrome (VCFS). Although the diagnosis of VCFS is primarily a clinical one, a test has been developed to identify the underlying chromosomal abnormality, ie, deletion of 22q11. OBJECTIVE: To describe characteristics and occurrence of the 22q11 deletion in a population with VIA. SETTING: Three tertiary referral centers. DESIGN: Retrospective case series of 23 patients with VIA who required intervention and had follow-up for more than 1 year. These patients' medical records were reviewed for indications for adenoidectomy, the presence of 22q11 deletion and whether a 22q11 deletion test was obtained, phenotypic evidence for VCFS, presence of a submucous cleft palate, velopharyngeal closure pattern, and type of speech intervention. RESULTS: Of the 23 patients, 9 underwent adenoidectomy for otitis media, 9 for obstructive sleep symptoms, and 5 for sinusitis therapy. Fourteen of the 23 patients were tested for a 22q11 deletion. Of these 14 patients, 9 had a 22q11 deletion with 5 having phenotypic evidence for VCFS. Six of the 23 patients had a submucous cleft palate, 2 of whom had a 22q11 deletion. CONCLUSIONS: Although VIA is uncommon, its occurrence should alert the otolaryngologist to the possibility of an underlying syndrome diagnosis. The 22q11 deletion test is beneficial in diagnosing patients with genotypic, but not phenotypic, VCFS in this population. In tested subjects of our patient population, 28% (4 patients) had the genotype for VCFS, without clinical evidence of VCFS. PMID- 10807334 TI - Appropriateness of routine postoperative chest radiography after tracheotomy. AB - OBJECTIVE: To determine the appropriateness of postoperative chest radiography after adult tracheotomy. DESIGN: Retrospective case series. SETTING: Tertiary care academic medical center. PATIENTS: The records of 379 consecutive adult patients who underwent tracheotomy by the Otolaryngology-Head and Neck Surgery Service from January 1992 to December 1996 were available for review and met inclusion criteria. All patients underwent postoperative chest radiography. MAIN OUTCOME MEASURES: Frequency of postoperative tracheotomy-associated complications, most significantly pneumothorax. RESULTS: The patients had no pneumothorax on postoperative chest films. Minor complications, which were found in 7.1% of the patients, included small bleeds, wound infection, and subcutaneous emphysema. Tracheostomy-associated death occurred in 2 patients (0.5%). CONCLUSIONS: Routine postoperative chest radiography is unnecessary after adult tracheotomy. Chest radiography may be indicated by clinically suspicious signs or symptoms. PMID- 10807335 TI - Hypothyroidism after treatment for nonthyroid head and neck cancer. AB - OBJECTIVES: To determine the incidence of posttreatment hypothyroidism in patients treated with surgery with or without radiotherapy for advanced-stage nonthyroid head and neck cancer and to make recommendations for its detection. DESIGN: A prospective study to assess the incidence and time frame of occurrence of hypothyroidism in patients by primary tumor site and treatment modality. Thyroid function tests were performed preoperatively, at the first postoperative visit, and then approximately every 6 months. Patients were followed up for up to 3 years. SETTING: Arthur G. James Cancer Hospital and Research Institute, Columbus, Ohio. PATIENTS: A total of 251 patients with nonthyroid head and neck cancer were originally enrolled; 198 patients with evaluable data were studied to determine the incidence of posttreatment hypothyroidism. Approximately 80% of the patients had advanced stage (III or IV) or recurrent cancer. RESULTS: The overall incidence of posttreatment hypothyroidism was 15% in 198 patients followed up for a mean of approximately 12 months. Hypothyroidism developed in 12% of patients treated with nonlaryngeal surgery and radiotherapy. The group undergoing total laryngectomy (with thyroid lobectomy) and radiotherapy had a 61% incidence of hypothyroidism. The average time to detection of hypothyroidism was 8.2 months. CONCLUSIONS: Approximately 15% of patients treated for advanced head and neck cancer with surgery and radiotherapy will develop hypothyroidism. Those treated with total laryngectomy and radiotherapy are at greatest risk. PMID- 10807336 TI - Green light photoplethysmography monitoring of free flaps. AB - BACKGROUND: Monitoring strategies have been developed to address the issue of detecting postoperative free flap ischemia in an effort to permit intervention and flap salvage. No one existing noninvasive method has been widely accepted in a clinical setting. Green light photoplethysmography (GLP) uses a diode to transmit green light into a tissue. Reflected light from hemoglobin in dermal capillary red blood cells is analyzed as light intensity along a frequency spectrum. A pure peak signal (1-2 Hz) is identified and provides a way to distinguish between perfused and nonperfused tissue. DESIGN: Prospective, blinded comparison. SUBJECTS: Sixty of 72 consecutive patients considered for free flap reconstruction were enrolled in a protocol to evaluate the efficacy of GLP. INTERVENTION: After free flap elevation, but before pedicle ligation, 120-second baseline measurements were obtained; 120-second measurements then occurred 5 minutes after the onset or release of individual venous or arterial occlusion. Signals were processed by fast Fourier transfer; a mean alternating current direct current (AC/ DC) ratio was cultivated for each signal. All data were analyzed in a blinded fashion. RESULTS: The AC/DC ratio of GLP was statistically significant across all flap perfusion states (P<.001). Each condition resulted in a unique GLP signal within 5 minutes of manipulation of each vessel. CONCLUSIONS: Green light photoplethysmography with AC/DC ratio analysis provides a rapid, precise method with which to determine flap ischemia and can differentiate venous compromised and arterial compromised flaps almost immediately after the onset of an ischemic insult. It may provide a clinically useful tool for postoperative free flap monitoring. PMID- 10807337 TI - Supracricoid partial laryngectomy for severe laryngeal stenosis. AB - We report 2 cases in which supracricoid partial laryngectomy and cricohyoidoepiglottopexy were used to restore the airway in cases of severe associated glottic and supraglottic laryngeal stenosis. PMID- 10807338 TI - Intraosseous ganglion of the temporomandibular joint presenting with otorrhea. AB - A ganglion cyst of the temporomandibular joint is a rare entity that commonly presents as a minimally tender, preauricular mass. This benign cystic lesion, which is lined by synovium and can be found in association with other joints, occasionally erodes adjacent bone to form an intraosseous ganglion. We discuss an unusual case of an intraosseous temporomandibular ganglion cyst that presented with bloody otorrhea. Examination revealed an external auditory canal mass with radiographic evidence of temporal bone erosion. The cyst was excised using a combined approach to the glenoid fossa and mastoid. The management principals of this case and a pertinent review of the literature are included. PMID- 10807339 TI - Lingual nerve injury during suspension microlaryngoscopy. AB - Lingual nerve injury is an uncommon complication of laryngoscopy. We report a case of isolated unilateral lingual nerve injury that occurred during suspension microlaryngoscopy. The injury was transient, with complete return of sensation within 3 months after surgery. Several mechanisms have been proposed to explain the occurrence of lingual nerve injury during laryngoscopy, including direct compression of the nerve caused by the laryngoscope, stretching of the nerve caused by cricoid pressure or instrumentation, and compression of the nerve between the medial and lateral pterygoid caused by manipulation of the mandible. The precise mechanism of injury in this case was not obvious, but stretching of the lingual nerve caused by pressure of the suspended laryngoscope on the tongue or retrolingual region was likely. The transient nature of the injury and the rapid return of the nerve to baseline function in this case are consistent with a neurapraxic injury. PMID- 10807341 TI - Quiz case 1. Gastric heterotopia. PMID- 10807340 TI - Adult herpetic laryngitis with concurrent candidal infection: a case report and literature review. AB - Rarely, adult herpetic laryngitis without involvement of the oropharynx has been reported. However, to our knowledge, laryngitis caused by herpes simplex virus with coexisting Candida albicans has not been reported. We report what we believe to be the first case of localized herpetic laryngitis superimposed by laryngeal Candida species infection in an immunosuppressed patient. This diagnosis was made on the basis of the findings of a laryngeal mucosal biopsy and ancillary testing using fungal stains and immunohistochemical stains for herpetic antigens. We also review the literature and discuss the clinical and diagnostic presentations, including potential pitfalls in the diagnosis. PMID- 10807342 TI - Quiz case 2. Cat-scratch disease (CSD). PMID- 10807343 TI - Quiz case 3. Traumatic ulcerative granuloma with stromal eosinophilia (TUGSE). PMID- 10807344 TI - Quiz case 4. Myxoma of the maxillary sinus. PMID- 10807345 TI - Preoperative coagulation studies prior to tonsillectomy. PMID- 10807346 TI - Coagulation studies prior to tonsillectomy: an unsettled and unsettling issue. PMID- 10807347 TI - A cost-effective approach for preoperative hemostatic assessment in children undergoing adenotonsillectomy. PMID- 10807348 TI - Antimicrobial treatment and recurrent rhinosinusitis. PMID- 10807349 TI - Laser-assisted endoscopic stapedectomy: a prospective study. AB - OBJECTIVE: To improve the techniques required to perform a stapedotomy without prosthesis (stapedioplasty). STUDY DESIGN: New infrared lasers were evaluated for potential use in otological surgery in guinea pigs. A prospective human trial of 34 primary stapes operations using the Argon ion laser was performed, with 11 stapedioplasties and 23 conventional stapedotomies as controls. METHODS: Laser tissue interactions were evaluated for temporal bone and live guinea pig tissues, measuring crater histology and labyrinthine temperature elevations. Patients undergoing stapedioplasty had Argon ion laser cuts with endoscopic assistance made in the anterior crus and footplate to mobilize the posterior segment of the stapes while the anterior portion remained fixed. RESULTS: Diode laser (808-nm) vaporization craters and temperature elevations in the vestibule were suitable for clinical use. Overall, stapedioplasty patients' hearing was improved with air bone gap closure to a mean of 8.3 dB (SD +/- 9.8 dB). CONCLUSIONS: Patients with anterior footplate otosclerosis are candidates for stapedioplasty preserving the annular ring and stapes tendon and eliminating prosthesis complications. High resolution small endoscopes, coupled with Argon ion or diode lasers promise to improve stapes visualization, enhancing the ability to perform minimally invasive surgery on the stapes footplate. PMID- 10807350 TI - Head and neck cancer: the importance of oxygen. AB - OBJECTIVES: To use recently introduced polarographic technology to characterize the distribution of oxygenation in solid tumors, explore the differences between severe hypoxia and true necrosis, and evaluate the ability to predict treatment outcomes based on tumor oxygenation. STUDY DESIGN: Prospective, nonrandomized trial of patients with advanced head and neck cancer, conducted at an academic institution. METHODS: A total of 63 patients underwent polarographic oxygen measurements of their tumors. Experiment 1 was designed to determine whether a gradient of oxygenation exists within tumors by examining several series of measurements in each tumor. Experiment 2 was an analysis of the difference in data variance incurred when comparing oxygen measurements using oxygen electrodes of two different sizes. Experiment 3 compared the proportion of tumor necrosis to the proportion of very low (< or =2.5 mm Hg) polarographic oxygen measurements. Experiment 4 was designed to explore the correlation between oxygenation and treatment outcomes after nonsurgical management. RESULTS: No gradient of oxygenation was found within cervical lymph node metastases from head and neck squamous cell carcinomas (P > .9). Tumor measurements achieved with larger (17 microm) electrodes displayed smaller variances than those obtained with smaller (12 microm) electrodes, although this difference failed to reach statistical significance (P = .60). There was no correlation between tumor necrosis and the proportion of very low (< or =2.5 mm Hg) oxygen measurements. There was a nonsignificant trend toward poorer locoregional control and overall survival in hypoxic tumors. CONCLUSIONS: Hypoxia exists within cervical lymph node metastases from head and neck squamous carcinomas, but the hypoxic regions are distributed essentially randomly. As expected, measurements of oxygen achieved with larger electrodes results in lowered variance, but with no change in overall tumor mean oxygen levels. Polarographic oxygen measurements are independent of tumor necrosis. Finally, oxygenation as an independent variable is incapable of predicting prognosis, probably reflecting the multifactorial nature of the biological behavior of head and neck cancers. PMID- 10807351 TI - Gene therapy for head and neck cancer. AB - OBJECTIVES/HYPOTHESIS: New treatment methods are needed for head and neck cancer to improve survival without increasing morbidity. Gene therapy is a potential method of improving patient outcome. Progress in gene therapy for cancer is reviewed with emphasis on the limitations of vector technology and treatment strategies. Given the current technological vector limitations in transmitting the therapeutic genes, treatments that require the fewest number of cells to be altered by the new gene are optimal. Therefore an immune-based gene therapy strategy was selected in which the tumors were transfected with the gene for an alloantigen, human leukocyte antigen (HLA)-B7, a class I major histocompatibility complex (MHC). This would restore an antigen presentation mechanism in the tumor to induce an antitumor response. This gene therapy strategy was tested in patients with advanced, unresectable head and neck cancer. STUDY DESIGN: Prospective trial. METHODS: Twenty patients with advanced head and neck cancer who had failed conventional therapy and did not express HLA-B7 were treated with gene therapy using a lipid vector by direct intratumoral injection. The gene therapy product contained the HLA-B7 gene and the beta2-microglobulin gene, which permits complete expression of the class I MHC at the cell surface. Patients were assessed for any adverse effects, for changes in tumor size, for time to disease progression, and for survival. Biopsy specimens were assessed for pathological response, HLA-B7 expression, apoptosis, cellular proliferation, CD-8 cells, granzyme, and p53 status. RESULTS: There were no adverse effects from the gene therapy. At 16 weeks after beginning gene therapy, four patients had a partial response and two patients had stable disease. Two of the tumors completely responded clinically, but tumor was still seen on pathological examination. The time to disease progression in the responding patients was 20 to 80 weeks. The median survival in patients who completed gene therapy was 54 weeks, compared with 21 weeks in patients whose tumors progressed after the first cycle of treatment. One patient survived for 106 weeks without any additional therapy. HLA B7 was demonstrated in the treated tumors, and increased apoptosis was seen in the responding tumors. CONCLUSION: Significant advances have been made in the field of gene therapy for cancer. Alloantigen gene therapy has had efficacy in the treatment of cancer and can induce tumor responses in head and neck tumors. Alloantigen gene therapy has significant potential as an adjunctive treatment of head and neck cancer. PMID- 10807352 TI - Effects of dietary restriction and antioxidants on presbyacusis. AB - OBJECTIVES/HYPOTHESIS: The premise of this study is that the membrane hypothesis of aging, also known as the mitochondrial clock theory of aging, is the basis for presbyacusis. Furthermore, it is proposed that treatment with antioxidants or dietary restriction can attenuate age-related hearing loss. Many studies have demonstrated a reduction in blood flow to specific tissues, including the cochlea, with aging. Hypoperfusion leads to the formation of reactive oxygen metabolites (ROM). ROM are highly toxic molecules that directly affect tissues including inner ear structures. In addition, ROM can damage mitochondrial DNA (mtDNA), resulting in the production of specific mtDNA deletions (mtDNA del4977 [human] or mtDNA del4834 [rat]; also known as the common aging deletion]. Previous corroborating data suggest that the common aging deletion mtDNA4834 may be associated not only with aging but also with presbyacusis, thus further strengthening the basis of the current studies. In this study, experiments provide compelling evidence that long-term treatment with compounds that block or scavenge reactive oxygen metabolites attenuate age-related hearing loss and reduce the impact of associated deleterious changes at the molecular level. STUDY DESIGN: Prospective randomized study. METHODS: One hundred thirty rats were randomly assigned to one of six groups with appropriate controls. Animals were divided into the following treatment arms: group 1, 30% caloric restriction; group 2, vitamin E oversupplementation; group 3, vitamin C over-supplementation; group 4, melatonin treatment; group 5, lazaroid treatment; and group 6, placebo. In addition, 10 animals were used to determine the appropriate caloric restriction. All subjects underwent baseline and every-3-month testing until their health failed (range, 18-28 mo; average, 25 mo). This testing included auditory sensitivity studies using auditory brainstem response (ABR) testing, as well as tissue analysis for mtDNA deletions using molecular biological techniques. At the conclusion of the study, animals underwent a final ABR test and were tested for mtDNA deletions in brain and inner ear tissues, and the opposite ear was used for histological analysis. RESULTS: Results indicated that the 30%-caloric-restricted group maintained the most acute auditory sensitivities, the lowest quantity of mtDNA deletions, and the least amount of outer hair cell loss. The antioxidant-treated subjects had improved auditory sensitivities, and a trend for fewer mtDNA deletions was observed compared with the placebo subjects. The placebo subjects had the poorest auditory sensitivity, the most mtDNA deletions, and the greatest degree of outer hair cell loss. CONCLUSIONS: Intervention designed to reduce reactive oxygen metabolite damage appears to protect against age-related hearing loss specifically and aging in general. This is reflected by an overall reduction in mtDNA deletions. These data also suggest that the common aging deletion appears to be associated with presbyacusis, as demonstrated by an increased frequency of the mtDNA del4834 in the cochleae with the most significant hearing loss. Nutritional and pharmacological strategies may very well provide rational treatment options that would limit the age-associated increase in ROM generation, reduce mtDNA damage, and reduce the degree of hearing loss as the organism advances in age. PMID- 10807353 TI - Laryngeal reinnervation with the hypoglossal nerve: II. Clinical evaluation and early patient experience. AB - OBJECTIVES/HYPOTHESIS: To determine whether the hypoglossal nerve (XII) can serve as a suitable donor for human laryngeal reinnervation. STUDY DESIGN: Prospective, nonrandomized. METHODS: Measurements were made on patients undergoing open neck procedures to determine the length of XII available and that required to perform XII-recurrent laryngeal nerve (RLN) anastomosis. The morbidity of combined XII and RLN injuries was studied using temporary lidocaine block of the ipsilateral XII in patients with unilateral vocal fold paralysis (UVFP). A pilot series of patients with UVFP who underwent XII-RLN reinnervation was evaluated for morbidity of the procedure, and for improvement in voice and swallowing. RESULTS: In 89 necks the average available length of XII was 2 cm less than that needed to reach the larynx, indicating the RLN stump must be at least 3 cm to allow tension free anastomosis. Twenty-five patients with untreated UVFP underwent temporary lidocaine block of XII; 8 had slight changes in their speech, none had increased aspiration. Nine patients underwent XII-RLN reinnervation. Postoperative speech analysis correlated well with the findings of the temporary lidocaine block of XII. One-year follow-up of five patients showed excellent voice quality, resolution of any preoperative aspiration, and minimal morbidity. Slight adductory movement of the reinnervated vocal fold was seen during tongue thrust. Electromyography confirmed substantial polyphasic action potentials in the thyroarytenoid muscle. CONCLUSIONS: The hypoglossal nerve is a very suitable donor for reinnervation of patients with UVFP. There should be enough length for primary XII-RLN anastomosis in most patients. Donor site morbidity is acceptable. Preoperative lidocaine block of XII is a good predictor of actual donor site morbidity and could be used to assess patients undergoing facial-hypoglossal anastomosis as well. PMID- 10807354 TI - Unaided speech in long-term tube-free tracheostomy. AB - OBJECTIVES/HYPOTHESIS: Objectives of the current study were as follows: 1) to determine whether a self-sustaining, tube-free tracheostomy with unaided speech capability can be achieved, 2) to propose new surgical techniques to enhance voluntary constriction of a tube-free tracheostoma to improve cough and speech production, and 3) to explain the mechanism involved in unaided speech after long term tracheostomy. The hypotheses were that long-term, tube-free tracheostomy could minimize the unwanted effects of conventional tube-dependent tracheotomy; that surgical shunting of the trachea to the skin surface can be established through a short, skin-lined, self-supporting, nostril-like opening; and that patients could generate speech and cough without the need to occlude the stoma or use stents and/or one-way valves if such a long-term tube-free tracheostoma were established. STUDY DESIGN: A prospective study of tube-free tracheostomy intended to establish unaided cough and speech. METHODS: Thirty-five patients who underwent long-term flap tracheostomies between 1992 and 1999 with the prospective intent of establishing a self-sustaining stoma without the need for tubes or stents were reviewed. The patients were trained to intentionally constrict the stoma for production of unaided speech and cough. The pertinent techniques developed for successful achievement of these characteristics are presented and reviewed. A new surgical technique using a local tendinous muscular sling was designed to further improve the efficacy of stomal constriction. RESULTS: A tube-free stoma was successfully established in all 35 patients. Eighteen patients achieved effective intentional constriction of the stoma and thereby were capable of unaided speech production. Four patients required an additional new surgical sling procedure to optimize unaided cough and speech production. Thirteen patients achieved only limited unaided speech production, but were satisfied to do nothing further. CONCLUSIONS: New surgical techniques and postoperative management to create a modified tube-free tracheostomy with self-constricting capabilities can enable production of effective cough and near normal generation of unaided speech. In this manner, the airway can be secured with little or no voice compromise. PMID- 10807355 TI - Intracranial extension of acquired aural cholesteatoma. AB - OBJECTIVE: Cholesteatoma of the petrous bone extending into the intracranial region is an unusual occurrence. Most cases have been attributed to secondary extension of a primary epidermal blastomatous malformation of the temporal bone into the middle or posterior fossae. Within the past two and a half decades, intracranial extension of acquired aural cholesteatoma has been recognized as a likely alternative to this mechanism. Recent literature has rejoined this observation by considering both primary and secondary cholesteatoma of the petrous bone as a single group, petrosal cholesteatoma. The present study is presented to analyze the clinical presentation, imaging findings, and surgical treatment of six patients with acquired aural cholesteatoma extending into the intracranial region. Findings in this study are compared with the extant literature on congenital and acquired cholesteatoma of the petrous bone. This study proposes that petrosal cholesteatoma is a valid anatomical construct; however, the pathogenesis of petrosal cholesteatoma is still important in understanding the clinical presentation and management of cholesteatoma that extends beyond the usual confines of the middle ear and mastoid. STUDY DESIGN: Retrospective case review conducted at a tertiary referral center. METHODS: From 1985 to 1999, 477 patients were surgically treated for acquired aural cholesteatoma. Patients with intracranial extension of cholesteatoma were studied. Clinical presentation, imaging studies, operative findings, surgical treatment, and postoperative results were evaluated. RESULTS: Six cases in a series of 477 patients with acquired aural cholesteatoma had intracranial extension of disease. In this series, the most frequent pathway for intracranial extension was supralabyrinthine through the supratubal recess into the middle cranial fossa. A less frequent pathway was via the retrofacial air cells into the posterior cranial fossa. Surgical access for removal of intracranial cholesteatoma was accomplished through several approaches including translabyrinthine, transcochlear, retrolabyrinthine, and middle cranial fossa. In two patients who had reoperation for possible residual disease, one was free of residual disease and one was found to have residual cholesteatoma in the region of the horizontal facial nerve. CONCLUSION: Acquired aural cholesteatoma can extend into either the middle or posterior cranial fossae. In this study, cholesteatoma extended into the middle fossa through the supratubal recess along the labyrinthine facial nerve and into or above the internal auditory canal. A less frequent path is through the retrofacial air cells into the posterior fossa. Intracranial acquired cholesteatoma is generally small and presents with complaints related to underlying otitis media rather than the neurological deficits that are often associated with primary petrous bone cholesteatoma. While computed tomography and magnetic resonance imaging are both required to differentiate congenital petrous cholesteatoma from other lesions of the petrous bone, computed tomography of the temporal bone is usually sufficient to diagnosis and define intracranial extension of acquired aural cholesteatoma. These lesions can be completely excised rather than exteriorized. PMID- 10807356 TI - Past history of otitis media and balance in four-year-old children. AB - OBJECTIVES/HYPOTHESIS: To obtain normative data for a population of children 4 years of age with respect to standard vestibular and balance test protocols and to determine, in the absence of concurrent middle ear effusion (MEE), the possible changes caused by a history of recurrent or persistent MEE. STUDY DESIGN: Comparative studies of the results of vestibular and balance tests in a cohort of young children with and without a history of MEE. METHODS: Seventy-one children, 4 years of age, with a well-documented history since early infancy regarding the presence or absence of MEE were evaluated using pneumatic otoscopy, tympanometry, audiometry, and vestibular and balance (rotational and moving platform posturography) tests. For the results of the vestibular and balance tests, comparisons were made between the group of 31 children (43.7%) without and the group of 40 children (56.3%) with a history of recurrent or persistent MEE, when a positive disease history was defined as at least a 10% cumulative percentage of time with MEE between early infancy and time of testing. RESULTS: When compared with children with a negative history of significant MEE, children with a positive history had a lower average gain to a rotational stimulus of 0.1 Hz, 150 degrees/s (0.57 vs. 0.44; P = .007). There were no significant differences between groups with respect to other measures. CONCLUSIONS: These results suggest that a history of recurrent or persistent MEE affects the vestibular and/or balance function of 4-year-old children when tested in the absence of a concurrent episode of MEE. The possible sequelae of the disease should be weighed in future considerations of early intervention for MEE. PMID- 10807357 TI - Hearing preservation with labyrinthine ablation in otitis media. AB - OBJECTIVES: Iatrogenic fenestration of the inner ear in the presence of otitis media is commonly associated with permanent hearing loss. Hearing can generally be preserved when the vestibular labyrinth is ablated in a controlled manner in noninflamed ears. The purpose of this study was to examine the feasibility of hearing preservation with violation of the inner ear in the presence of middle ear inflammation. STUDY DESIGN: Prospective and controlled animal model. METHODS: Otitis media was induced bilaterally in pigmented guinea pigs with transtympanic injection of Streptococcus pneumoniae, nontypeable Haemophilus influenzae, or formalin-killed nontypeable H influenzae. Two to 4 days after injection, the horizontal canal of one ear was transected and sealed. Hearing was tested before and after labyrinthine ablation. RESULTS: Otitis media was induced in all ears. Bacterial cultures were positive in 19 of 20 S pneumoniae-injected ears, and in 10 of 16 nontypeable H influenzae-injected ears. One week after surgery, elevation of click thresholds (> 15 dB) was encountered in none of the fenestrated or unfenestrated S pneumoniae-infected ears, in two of six unfenestrated and three of six fenestrated nontypeable H influenzae-infected ears, and in one of five killed-nontypeable H influenzae-injected ears both with and without fenestration. CONCLUSIONS: These data suggest that ablation of a semicircular canal in the presence of middle ear inflammation or infection does not necessarily lead to profound sensorineural hearing loss. Hearing loss associated with iatrogenic violation of the semicircular canals may be more dependent on factors other than the presence of nonspecific middle ear inflammation. PMID- 10807358 TI - Analysis of a cohort of children with sensory hearing loss using the SCALE systematic nomenclature. AB - OBJECTIVES: What characteristics identify clinical types of childhood deafness? Which aspects of the otological evaluation best delineate them? To approach these related questions, a classification for deafness consistent with current medical concepts was constructed using a systematic nomenclature and then applied to a pediatric cohort of 168 children with sensorineural hearing loss (SNHL) who were referred for private consultation. A major aim of the analysis was to test the utility of SCALE, the new systematic nomenclature. METHODS: Patients with SNHL were identified through the office records of a single faculty member of the Department of Otolaryngology in a medical school situated in a major US city. Inclusion criteria required bone conduction thresholds above 30 dB or equivalent in at least one of the frequencies from 250 Hz to 4 kHz on either behavioral audiogram or on electrophysiological testing. All identified patients had initial visits during an 8-year period from late 1990 to early 1999. Patients were excluded if age at first consultation was 19 years or more, if records were insufficient to confirm SNHL, or if further evaluation revealed that SNHL had been misdiagnosed. A formal nomenclature was designed to systematically encode clinical features with simple descriptive terms according to an acronym (SCALE [sidedness, component function, age of onset, lesion, and etiology]) for all included patients. RESULTS: One hundred sixty-eight study patients were analyzed; sensory hearing loss was bilateral in 82% (137/168) and unilateral in 18% (31/168). The etiology of this impairment was determined to be intrinsic in 40% of children (67/168), either secondary to genotype (57/ 67), or to named congenital syndromes without known extrinsic cause (10/67). Recessive single gene mutations diagnosed by family history, recognition of syndrome, or determination of homozygous 35delG mutations in the gap junction protein gene, Connexin 26, accounted for bilateral sensory hearing loss in 33 children (24% of all bilateral cases). One girl had an X-linked dominant syndrome (Coffin-Lowry syndrome) with auditory brainstem response-documented childhood onset of SNHL. Nine patients (5%) had chromosomal aneuploidy, and 12 patients (7%) had either a family history of dominant deafness (7/ 12) or a recognizable autosomal dominant syndrome (5/12), most commonly, Waardenburg syndrome type 1 (4/5). Extrinsic causes of deafness were identified in only 13% of children (21/168) and included a relatively large number of referrals from neurosurgery (9/21). Three of these children had chronic middle ear disease and sensory hearing loss associated with inflammatory and bony changes on temporal bone imaging suggestive of chronic osteitis; all had a history of active otitis media during cranial irradiation. Congenital cytomegalovirus infections were documented in only 4 cases, but 41 patients could have had this as a cause or did not have this cause ruled out. An idiopathic cause or origin was assigned to 36% of patients (61/168) including patients with unnamed syndromic patterns of multiple anomalies. CONCLUSIONS: The SCALE nomenclature facilitated complete descriptions of hearing-impaired children and provided a classification scheme applicable to broad categories of human disease. The single most useful diagnostic test was screening for Cx26 mutations. Computed tomography scan of the temporal bones was helpful in establishing etiology for selected patients and invaluable in patients with chronic ear disease. Magnetic resonance imaging scan was a superior diagnostic modality in one child with a posterior fossa arachnoid cyst. PMID- 10807359 TI - Eosinophilic mucin rhinosinusitis: a distinct clinicopathological entity. AB - BACKGROUND: Allergic fungal sinusitis (AFS) was recognized in 1981. Since 1983, a form of sinusitis histologically similar to AFS except for the absence of fungal hyphae has also been noted. The designation "eosinophilic mucin rhinosinusitis (EMRS)" is proposed. Its relationship to AFS is controversial and problematic. OBJECTIVE: To determine whether distinctive clinical and immunological differences exist to differentiate the histological entity of EMRS from AFS. STUDY DESIGN: Literature review and comparison of cases of AFS (n = 418) to EMRS (n = 40) from the literature, as well as cases of AFS (n = 13) and EMRS (n = 29) accrued in the present study. RESULTS: A total of 431 AFS patients were compared with 69 EMRS patients. The mean age of patients with AFS was significantly younger than patients with EMRS (30.7 y compared with 48.0 y, respectively; P < .001). Male-to-female ratios were 1.03:1 and 1.26:1 for AFS and EMRS, respectively, and were not significantly different. Forty-one percent of patients with AFS were asthmatic compared with 93% of patients with EMRS (P < .0001). Thirteen percent of patients with AFS were aspirin sensitive compared with 54% of patients with EMRS (P < .0001). Polyp occurrence was almost 100% in both groups. Eighty-four percent of patients with AFS had allergic rhinitis (AR), while only 63% of patients with EMRS had AR (P = .004). Fifty-five percent of AFS patients had bilateral disease, in contrast to the 100% of EMRS patients with bilateral disease (P < .0001). Although average total immunoglobulin E (IgE) was elevated in both groups, it was significantly more elevated in AFS patients (range, 12 13,084 mg/ dL; mean, 1,941 mg/dL) compared with EMRS patients (range, 14-1,162 mg/dL; mean, 267 mg/dL; P < .001). Total immunoglobulin G (IgG) and IgG subclasses were seldom reported in the cases available from the literature of either AFS or EMRS. However, in the present series of EMRS, IgG1 deficiency occurred in 50% of evaluated patients (mean, 475 +/- 175 mg/dL; range, 250-869 mg/dL; normal, 422 to 1,200 mg/dL) but in no cases of AFS reported in the literature. CONCLUSIONS: Significant clinical and immunological differences exist to distinguish AFS from EMRS. It is postulated that AFS is an allergic response to fungi in predisposed individuals, while EMRS occurs because of a systemic dysregulation of immunological controls. Because EMRS is a systemic disease, unilateral disease is not seen. In contrast, AFS, an allergic response to fungi, may occur unilaterally or bilaterally depending on the antigenic stimulation. EMRS also has a significantly higher association with asthma, an increased incidence of aspirin sensitivity, and an increased incidence of IgG1 deficiency. Therapy with a systemic steroid, a potent and indiscriminant anti-inflammatory agent, is a useful adjunct in both disorders. Fungal immunotherapy following surgical extirpation of AFS is useful in preventing AFS recurrence. It is predicted that fungal immunotherapy and antifungal agents will be ineffective in patients with EMRS. It is important to differentiate these two similar histopathological entities in future trials assessing therapeutic efficacy. Inclusion of both entities in a study could obscure recognition of the true effectiveness of intervention, because of the possible variable response differences between the two entities. This study shows that significant clinical and immunological differences exist between EMRS and AFS. The future awaits an exploration of the pathophysiological basis of these differences. PMID- 10807360 TI - Comparative histology and vibration of the vocal folds: implications for experimental studies in microlaryngeal surgery. AB - OBJECTIVES/HYPOTHESIS: To determine the most suitable animal model for experimental studies on vocal fold surgery and function by a histological comparison of the microflap surgical plane and laryngeal videostroboscopy (LVS) in different species of animals. A second goal was to determine how the layered vocal fold structure in humans and three different animal species affects surgical dissection within the lamina propria. STUDY DESIGN: Prospective laboratory. METHODS: Three larynges each from dogs, monkeys, and pigs were compared with three ex vivo human larynges. Microflap surgery was performed on one vocal fold from each larynx. Both the operated and nonoperated vocal folds were examined histologically using stains specific for elastin, mature collagen, and ground substance. Based on the histological results, LVS was performed on two dogs and two pigs after first performing a tracheotomy for ventilation and airflow through the glottis. Arytenoid adduction sutures were placed to facilitate vocal fold adduction. RESULTS: The distributions of the collagen and elastin fibers were found to differ among the species with concentrations varying within species. Unlike the human vocal fold, which has a higher elastin concentration in the deeper layers of the lamina propria, both the pig and the dog had a thin band of elastin concentrated just deep to the basement membrane zone in the superficial layer. Just deep to this thin band, the collagen and the elastin were less concentrated. The monkey vocal fold had a very thin mucosal layer with less elastin throughout the mucosa. The microflap dissections in each of the dog, pig, and human vocal folds were similar, being located within that portion of the superficial lamina propria where the elastin and mature collagen are less concentrated. The microflap plane in the monkey vocal fold was more deeply located near the vocalis fibers. Despite the differences in elastin concentration, the microflap plane in both the dog and the pig was found to be similar to that in humans. The dog anatomy was much more suitable for microsuspension laryngoscopy and stroboscopic examination. The dog vocal folds vibrated in a similar fashion to human vocal folds with mucosal waves and vertical phase differences, features not seen in the pig vocal folds. CONCLUSIONS: Based on both the histological and stroboscopic results, the dog was believed to be a more suitable animal model for studies on vocal fold surgery, acknowledging that no animal's laryngeal anatomy is identical to that of the human. The dog LVS model presented allows for longitudinal laryngeal studies requiring repeated examinations at multiple time periods with histological correlation applied at sacrifice. PMID- 10807361 TI - Benefit of feeding assessment before pediatric airway reconstruction. AB - OBJECTIVES/HYPOTHESIS: To determine the utility of preoperative feeding assessments in children undergoing airway reconstruction, identifying parameters that correlate with functional deficits in swallowing and postoperative feeding difficulties. STUDY DESIGN: Prospective, sequential enrollment. METHODS: Two hundred fifty-five patients with structural abnormalities of the upper aerodigestive tract underwent endoscopic swallow studies. Classification of preoperative feeding abilities, specific feeding disorders, and abnormal endoscopic feeding parameters were used to predict the postoperative course of patients undergoing airway reconstruction. The relationships between diagnoses and functional feeding categories and postoperative outcomes and functional feeding categories were appraised by chi2 analysis. RESULTS: The median age of the study population was 2.5 years. Fifty-three percent of the patients were tracheotomy dependent. Only 13% of the patients had diagnoses limited to the airway, with 45% of patients having three or more diagnoses. Worse preoperative feeding abilities were associated with the presence of a tracheotomy, age 2 years or less, and multiple underlying diagnoses. Neurological diagnoses were associated with worse feeding abilities. Preoperative feeding assessments directly altered the course of management of 15% of operative patients, by recommending a delay in the surgical correction, the placement of a gastrostomy tube preoperatively, or a modification in the surgical reconstruction planned for the patient. Postoperative airway protection predictions were 80% accurate. Twelve percent of the predictions involved patients who developed unforeseen complications that required additional treatments or prolonged the hospital stay secondary to difficulties with airway protection. There was no correlation between the preoperative feeding abilities of the patients and their postoperative course after airway reconstruction. CONCLUSION: Transient dysphagia is common after laryngotracheal reconstruction. Preoperative feeding abilities do not correlate with the postoperative airway protection abilities of a patient. Feeding assessments before pediatric airway reconstruction provide a means of identifying patients with poor airway protection mechanisms that may compromise the patient after reconstruction. Findings on swallowing evaluations that predict poor airway protective mechanisms are 1) pooling of secretions in the hypopharynx, 2) poor oral motor skills, allowing premature spillage of material into the hypopharynx where it penetrates the larynx, and 3) residue that persists in the hypopharynx after multiple swallows. The integration of information generated from the preoperative swallowing assessment promotes the selection of operative procedures that are optimal for that patient and highlights specific therapy issues that may need to be addressed in the postoperative management of the patient that may not have been obvious without the study. PMID- 10807362 TI - Airway growth after cricotracheal resection in a rabbit model and clinical application to the treatment of subglottic stenosis in children. AB - OBJECTIVE: To evaluate the effects of a partial resection of the anterior cricoid arch on the growth of the laryngotracheal complex in an animal model and to study the lumen caliber and cross sectional surface area following resection as the animals are allowed to grow. Laryngeal, subglottic, and tracheal measurements in the infant and pediatric larynges in autopsy specimens are also investigated. DESIGN: A controlled animal study was done comparing the operated rabbit group to the nonoperated group. Twenty-nine infant rabbits were compared to a control group of ten rabbits. In addition, autopsy dissections were performed on infant and pediatric larynges to obtain measurements of various laryngeal and cricoid relationships. METHODS: Thirty-nine pasteurella-free New Zealand white female rabbits between the ages of 8 and 11 weeks were used in this study. In the experimental group, cricotracheal resections were performed by removing the anterior cricoid cartilage and the first tracheal ring. A primary anastomosis was then performed. The animals were kept alive for 4 months and then humanely euthanized. The laryngeal complex was then removed and histological sections of the cricotracheal region were mounted on glass slides and stained with H&E. Measurements were taken of the cricoid and upper tracheal lumen and cross sectional area. RESULTS: Twenty-seven of the study group survived until the time of sacrifice. All of the control group survived during the study period. The two groups were comparable in regard to weight, lumen size, and cross-sectional area There were no statistical differences between the groups. CONCLUSIONS: The experimental data support the hypothesis that partial cricotracheal resections can be performed safely and effectively in young rabbits. The potential clinical significance is described in light of the autopsy data. PMID- 10807363 TI - A functional model system of an hypoxic nerve injury and its evaluation. AB - OBJECTIVES/HYPOTHESIS: Develop an hypoxic peripheral nerve injury model with a controlled injury type and two types of clinically relevant physiological measurements of function during and after recovery. The model, controlling for injury and measurement variables, would have predictable outcomes in function. The functional model could test potential therapeutic interventions with greater sensitivity. STUDY DESIGN: Twenty-one rats were used in preliminary studies evaluating peroneal nerve injury types and functional model evaluation. Forty eight rats were used in a controlled and blinded evaluation of the injury model followed by treatment with hyperbaric oxygen (HBO) as a potential therapeutic intervention and evaluated with functional models. METHODS: Preliminary studies compared nerve injuries: epineurectomy, epineurectomy with crush and transection with autograft for rate of return of function and final extent of return of function. The gait analysis model was also evaluated and modified to decrease variability. The final study evaluated peroneal epineurectomy and nerve crush injury with serial gait analysis during recovery, final elicited maximum force measurements, and histological analysis. Half of the animals were treated with HBO during recovery (ANOVA or regression statistical analysis were used to determine group differences.). RESULTS: Preliminary studies suggested that the peroneal nerve injury model of an epineurectomy with crush of specified length and a modification of the gait analysis model would yield a useful and predictable injury outcome. The final study resulted in predicted and consistent injury outcomes. In the HBO treatment group, a 12% improvement in function 5 days after HBO treatment was demonstrated (P < .03), but no long-term or histological benefit was seen. CONCLUSION: A reliable hypoxic nerve injury model has been developed and tested utilizing two functional methods as the primary outcome variables. PMID- 10807364 TI - The versatile contact Nd:YAG laser in head and neck surgery: an in vivo and clinical analysis. AB - OBJECTIVE: Lasers have been used in otolaryngology as a surgical instrument for more than 25 years, and the CO2 laser has emerged as the most widely employed surgical laser in use today. However, recent technological advances have made the Nd:YAG laser a challenger as an effective photothermal surgical tool. STUDY DESIGN AND METHODS: This is a two-part study. Tissue injury and healing profiles after application of both the CO2 and Nd:YAG lasers are compared using an in vivo rat tongue model. A prospective clinical review based on the experience of 327 operative cases spanning a 7-year interval using the Nd:YAG laser, highlighting its various applications and associated complications, is detailed. RESULTS: Comparable tissue and healing effects were noted with both lasers in the in vivo rat tongue model with no statistical differences. The clinical application of the laser showed wide versatility in the head and neck with a complication rate of 3%. CONCLUSION: The Nd:YAG laser has proved equivalent in tissue damage and healing to the CO2 laser. The Nd:YAG laser has proved itself to be an excellent and perhaps superior laser for use in head and neck surgery. PMID- 10807365 TI - Epidermal growth factor receptor--mediated stat3 signaling blocks apoptosis in head and neck cancer. AB - OBJECTIVES: Upregulation of epidermal growth factor receptor (EGFR) is critical for the loss of growth control in a variety of human cancers including squamous cell cancers of the head and neck (SCCHN). In these tumor cells in culture, EGFR stimulation initiates signaling via persistent activation of STAT proteins, particularly Stat3. The present study was conducted to study the association between EGFR stimulation and constitutive activation of Stat3 in SCCHN in vivo and to investigate the proliferative and apoptotic consequences of Stat3 downmodulation in SCCHN cells in vitro. METHODS: SCCHN tumor xenografts were analyzed using electrophoretic mobility shift assay. A dominant-negative mutant Stat3 expression construct or a Stat3 antisense plasmid was transfected into SCCHN cells using lipofectamine. Cell growth and apoptosis were determined by vital dye exclusion and flow cytometry, respectively. RESULTS: In vivo liposome mediated gene therapy with an EGFR antisense plasmid efficiently inhibited Stat3 activation in a head and neck xenograft model. Downmodulation of Stat3 using a dominant-negative or antisense approach inhibited tumor cell growth and stimulated apoptosis. CONCLUSIONS: These findings provide evidence that constitutively activated Stat3 is linked to EGFR signaling in SCCHN in vivo, which contributes to the loss of growth control by an anti-apoptotic mechanism. PMID- 10807366 TI - Dermatologists debate sentinel node biopsy, safety of liposuction, and antibiotic prophylaxis. PMID- 10807367 TI - NIDA seeking data on effect of fetal exposure to methamphetamine. PMID- 10807368 TI - Travelers need hepatitis and other immunizations. PMID- 10807370 TI - From the Centers for Disease Control and Prevention. Salmonellosis associated with chicks and ducklings--Michigan and Missouri, spring 1999. PMID- 10807369 TI - From the Food and Drug Administration. PMID- 10807371 TI - From the Centers for Disease Control and Prevention. Fatal yellow fever in a traveler returning from Venezuela, 1999. PMID- 10807372 TI - From the Centers for Disease Control and Prevention. Hantavirus pulmonary syndrome--Panama, 1999-2000. PMID- 10807373 TI - From the Centers for Disease Control and Prevention. Motor-vehicle occupant fatalities and restraint use among children aged 4-8 years--United States, 1994 1998. PMID- 10807374 TI - Chronic systemic inflammation in overweight and obese adults. PMID- 10807375 TI - Chronic systemic inflammation in overweight and obese adults. PMID- 10807376 TI - Raloxifene and risk of vertebral fracture in postmenopausal women. PMID- 10807377 TI - Treatment decisions for seriously ill newborns. PMID- 10807378 TI - Treatment decisions for seriously ill newborns. PMID- 10807379 TI - Sexual function after hysterectomy. PMID- 10807380 TI - Treatment of postoperative respiratory failure. PMID- 10807381 TI - Circadian relationship of serum uric acid and nitric oxide. PMID- 10807382 TI - Alcohol and motor vehicle-related deaths of children as passengers, pedestrians, and bicyclists. AB - CONTEXT: The overall percentage of motor vehicle deaths associated with alcohol consumption declined between 1991 and 1996, but the risk of death due to alcohol related crashes for children warrants analysis. OBJECTIVE: To examine the association between alcohol use by drivers and mortality of children who were passengers, pedestrians, and bicyclists. DESIGN AND SETTING: Analysis of data from the Fatality Analysis Reporting System, a nationwide US registry of motor vehicle deaths, for 1991-1996. SUBJECTS: A total of 16,676 children younger than 16 years who were passengers, pedestrians, or bicyclists and whose death was due to a motor vehicle crash. MAIN OUTCOME MEASURE: Alcohol use by drivers involved in crashes in which children died, assessed by age and sex of the child and driver and type of crash. RESULTS: A total of 3310 deaths (19.9%) involved alcohol-related crashes. The percentage declined from 21.6% in 1991 to 17.8% in 1996. Considering only crashes in which the alcohol-use status of the child's driver was relevant, the decline was less marked, from 18.8% in 1991 to 15.1% in 1995, with an increase to 16.4% in 1996. Among crashes involving alcohol, the child's own driver had been drinking in 66.3% of cases, varying from 58.0% to 70.7% over time. Drivers younger than the legal drinking age of 21 years who had been drinking alcohol accounted for 30.3% of alcohol-related passenger deaths among children. CONCLUSION: While the overall percentage of alcohol-related motor vehicle deaths for children declined between 1991 and 1996, experiences for passengers, pedestrians, and bicyclists differ. Selected characteristics of children and drivers that elevate the risk of an alcohol-related motor vehicle death point to the need for further policy and clinical interventions. PMID- 10807383 TI - Characteristics of child passenger deaths and injuries involving drinking drivers. AB - CONTEXT: Motor vehicle-related injury is the leading cause of death for children and young adults aged 1 to 24 years in the United States. Approximately 24% of child traffic deaths involve alcohol. OBJECTIVE: To examine characteristics of crashes involving child passenger deaths and injuries associated with drinking drivers to identify opportunities for prevention. DESIGN, SETTING, AND PARTICIPANTS: Descriptive epidemiological analysis of 1985-1996 datafrom the Fatality Analysis Reporting System on deaths among US child passengers (aged 0-14 years) and 1988-1996 data from the General Estimates System on nonfatal injuries. MAIN OUTCOME MEASURES: Child passenger death or injury by driver characteristics (eg, driver age, blood alcohol concentration, and driving history). RESULTS: In 1985-1996, there were 5555 child passenger deaths involving a drinking driver. Of these deaths, 3556 (64.0%) occurred while the child was riding with a drinking driver; 67.0% of these drinking drivers were old enough to be the parent or caregiver of the child. Of all drivers transporting a child who died, drinking drivers were more likely than nondrinking drivers to have had a previous license suspension (17.1% vs 7.1%) or conviction for driving while intoxicated (7.9% vs 1.2%). Child restraint use decreased as both the child's age and the blood alcohol concentration of the child's driver increased. In 1988-1996, an estimated 149,000 child passengers were nonfatally injured in crashes involving a drinking driver. Of these, 58,000 (38.9%) were riding with a drinking driver when injured in the crash. CONCLUSIONS: These data indicate that the majority of drinking driver-related child passenger deaths in the United States involve a child riding unrestrained in the same vehicle with a drinking driver. Typically, the drinking driver transporting the child is old enough to be the child's parent or caregiver. PMID- 10807384 TI - Incident type 2 diabetes mellitus in African American and white adults: the Atherosclerosis Risk in Communities Study. AB - CONTEXT: Although the excess prevalence of type 2 diabetes mellitus in African Americans is well established, few studies have compared incident diabetes in African American and white persons. OBJECTIVES: To compare risk of incident diabetes in African American vs white adults and to identify explanatory factors for racial disparities. DESIGN: Prospective cohort study using baseline data collected from 1986 to 1989 from the ongoing Atherosclerosis Risk in Communities (ARIC) Study, with 9 years of follow-up. SETTING AND PARTICIPANTS: A total of 2646 African American and 9461 white adults aged 45 to 64 years without diabetes at baseline, sampled from 4 US communities. MAIN OUTCOME MEASURES: Incident type 2 diabetes, ascertained by self-report of physician diagnosis, use of diabetes medications, or fasting glucose level of at least 7.0 mmol/L (126 mg/dL), compared among white and African American subjects and by presence of potentially modifiable risk factors. RESULTS: Diabetes incidence per 1000 person-years was about 2.4-fold greater in African American women (25.1 [95% confidence interval [CI], 22.4-28.1] vs 10.4 [95% CI, 9.4-11.4]) and about 1.5-fold greater in men (23.4 [95% CI, 19.9-27.2] vs 15.9 [95% CI, 14.6-17.2]) than in their white counterparts (P<.001). Results from proportional hazards regression models indicated that racial differences in potentially modifiable risk factors, particularly adiposity, accounted for 47.8% of the excess risk in African American women but accounted for little excess risk in African American men. Compared with their white counterparts, African American men and women had higher blood pressures before diabetes onset (diastolic blood pressure difference=5.6 mm Hg in women and 8.4 mm Hg in men; P=.005). CONCLUSIONS: Our data indicate that compared with their white counterparts, middle-aged African Americans are at greater risk of developing type 2 diabetes and have higher blood pressure prior to development of diabetes. In women, almost 50% of this excess risk might be related to potentially modifiable factors. PMID- 10807385 TI - Clinicopathologic features of BRCA-linked and sporadic ovarian cancer. AB - CONTEXT: Most hereditary ovarian cancers are associated with germline mutations in BRCA1 or BRCA2. Attempts to define the clinical significance of BRCA mutation status in ovarian cancer have produced conflicting results, especially regarding survival. OBJECTIVE: To determine whether hereditary ovarian cancers have distinct clinical and pathological features compared with sporadic (nonhereditary) ovarian cancers. DESIGN AND SETTING: Retrospective cohort study of a consecutive series of 933 ovarian cancers diagnosed and treated at our institution, which is a comprehensive cancer center as designated by the National Cancer Institute, over a 12-year period (December 1986 to August 1998). PATIENTS: The study was restricted to patients of Jewish origin because of the ease of BRCA1 and BRCA2 genotyping in this ethnic group. From the 189 patients who identified themselves as Jewish, 88 hereditary cases were identified with the presence of a germline founder mutation in BRCA1 or BRCA2. The remaining 101 cases from the same series not associated with a BRCA mutation and 2 additional groups (Gynecologic Oncology Group protocols 52 and 111) with ovarian cancer from clinical trials (for the survival analysis) were included for comparison. MAIN OUTCOME MEASURES: Age at diagnosis, surgical stage, histologic cell type and grade, and surgical outcome; and response to chemotherapy and survival for advanced-stage (II and IV) cases. RESULTS: Hereditary cancers were rarely diagnosed before age 40 years and were common after age 60 years, with mean age at diagnosis being significantly younger for BRCA1- vs BRCA2-linked patients (54 vs 62 years; P=.04). Histology, grade, stage, and success of cytoreductive surgery were similar for hereditary and sporadic cases. The hereditary group had a longer disease-free interval following primary chemotherapy in comparison with the nonhereditary group, with a median time to recurrence of 14 months and 7 months, respectively (P<.001). Those with hereditary cancers had improved survival compared with the nonhereditary group (P=.004). For stage III cancers, BRCA mutation status was an independent prognostic variable (P=.03). CONCLUSIONS: Although BRCA-associated hereditary ovarian cancers in this population have surgical and pathological characteristics similar to those of sporadic cancers, advanced-stage hereditary cancer patients survive longer than nonhereditary cancer patients. Age penetrance is greater for BRCA1-linked than for BRCA2-linked cancers in this population. PMID- 10807386 TI - Respiration during snow burial using an artificial air pocket. AB - CONTEXT: Asphyxia is the most common cause of death after avalanche burial. A device that allows a person to breathe air contained in snow by diverting expired carbon dioxide (CO2) away from a 500-cm3 artificial inspiratory air pocket may improve chances of survival in avalanche burial. OBJECTIVE: To determine the duration of adequate oxygenation and ventilation during burial in dense snow while breathing with vs without the artificial air pocket device. DESIGN: Field study of physiologic respiratory measures during snow burial with and without the device from December 1998 to March 1999. Study burials were terminated at the subject's request, when oxygen saturation as measured by pulse oximetry (SpO2) dropped to less than 84%, or after 60 minutes elapsed. SETTING: Mountainous outdoor site at 2385 m elevation, with an average barometric pressure of 573 mm Hg. PARTICIPANTS: Six male and 2 female volunteers (mean age, 34.6 years; range, 28-39 years). MAIN OUTCOME MEASURES: Burial time, SpO2, partial pressure of end tidal CO2 (ETCO2), partial pressure of inspiratory CO2 (PICO2), respiratory rate, and heart rate at baseline (in open atmosphere) and during snow burial while breathing with the device and without the device but with a 500-cm3 air pocket in the snow. RESULTS: Mean burial time was 58 minutes (range, 45-60 minutes) with the device and 10 minutes (range, 5-14 minutes) without it (P=.001). A mean baseline SpO2 of 96% (range, 90%-99%) decreased to 90% (range, 77%-96%) in those buried with the device (P=.01) and to 84% (range, 79%-92%) in the control burials (P=.02). Only 1 subject buried with the device, but 6 control subjects buried without the device, decreased SpO2 to less than 88% (P=.005). A mean baseline ETCO2 of 32 mm Hg (range, 27-38 mm Hg) increased to 45 mm Hg (range, 32-53 mm Hg) in the burials with the device (P=.02) and to 54 mm Hg (range, 44-63 mm Hg) in the control burials (P=.02). A mean baseline PICO2 of 2 mm Hg (range, 0-3 mm Hg) increased to 32 mm Hg (range, 20-44 mm Hg) in the burials with the device (P=.01) and to 44 mm Hg (range, 37-50 mm Hg) in the control burials (P=.02). Respiratory and heart rates did not change in burials with the device but significantly increased in control burials. CONCLUSIONS: In our study, although hypercapnia developed, breathing with the device during snow burial considerably extended duration of adequate oxygenation compared with breathing with an air pocket in the snow. Further study will be needed to determine whether the device improves survival during avalanche burial. PMID- 10807387 TI - Radiation exposure from outpatient radioactive iodine (131I) therapy for thyroid carcinoma. AB - CONTEXT: In May 1997, the US Nuclear Regulatory Commission (NRC) revised its patient release regulations, allowing for outpatient administration of larger activities of sodium iodide 131I than previously permitted. OBJECTIVE: To measure the radiation exposure to household members from patients receiving outpatient 131I therapy for thyroid carcinoma in accordance with the new regulations. DESIGN: Consecutive case series from October 1998 to June 1999. SETTING AND PATIENTS: Thirty patients who received outpatient 131I therapy following thyroidectomy for differentiated thyroid carcinoma were enrolled, along with their 65 household members and 17 household pets. MAIN OUTCOME MEASURE: Radiation exposure to household members and 4 rooms in each home, as monitored with dosimeters for 10 days following 131I administration. RESULTS: The patients received 131I doses ranging from 2.8 to 5.6 GBq (mean, 4.3 GBq). The radiation dose to 65 household members ranged from 0.01 mSv to 1.09 mSv (mean, 0.24 mSv). The dose to 17 household pets ranged from 0.02 mSv to 1.11 mSv (mean, 0.37 mSv). The mean dose to the 4 rooms ranged from 0.17 mSv (kitchen) to 0.58 mSv (bedroom). CONCLUSION: In our study, 131I doses to household members of patients receiving outpatient 131I therapy were well below the limit (5.0 mSv) mandated by current NRC regulations. PMID- 10807388 TI - Determining when quality improvement initiatives should be considered research: proposed criteria and potential implications. AB - Quality improvement initiatives have become a major force in shaping health care. These initiatives are diverse but share a common goal of generating knowledge that will guide improvements in health care. Although quality improvement occupies an uncertain territory between clinical care and research, this difference is extremely important. If a quality improvement initiative is research, federal regulations require a variety of procedures that must be followed to protect the human subjects involved. Conversely, if the same initiative is not research, the regulations outlining these procedures are not applicable. This article proposes 2 criteria to determine whether a quality improvement initiative should be reviewed as research. Such an initiative should be reviewed and regulated as research if (1) the majority of patients involved are not expected to benefit directly from the knowledge to be gained or (2) if additional risks or burdens are imposed to make the results generalizable. Implementation of these criteria offers the possibility that the rights and interests of those who participate in quality improvement initiatives will be protected. PMID- 10807389 TI - Plague as a biological weapon: medical and public health management. Working Group on Civilian Biodefense. AB - OBJECTIVE: The Working Group on Civilian Biodefense has developed consensus-based recommendations for measures to be taken by medical and public health professionals following the use of plague as a biological weapon against a civilian population. PARTICIPANTS: The working group included 25 representatives from major academic medical centers and research, government, military, public health, and emergency management institutions and agencies. EVIDENCE: MEDLINE databases were searched from January 1966 to June 1998 for the Medical Subject Headings plague, Yersinia pestis, biological weapon, biological terrorism, biological warfare, and biowarfare. Review of the bibliographies of the references identified by this search led to subsequent identification of relevant references published prior to 1966. In addition, participants identified other unpublished references and sources. Additional MEDLINE searches were conducted through January 2000. CONSENSUS PROCESS: The first draft of the consensus statement was a synthesis of information obtained in the formal evidence gathering process. The working group was convened to review drafts of the document in October 1998 and May 1999. The final statement incorporates all relevant evidence obtained by the literature search in conjunction with final consensus recommendations supported by all working group members. CONCLUSIONS: An aerosolized plague weapon could cause fever, cough, chest pain, and hemoptysis with signs consistent with severe pneumonia 1 to 6 days after exposure. Rapid evolution of disease would occur in the 2 to 4 days after symptom onset and would lead to septic shock with high mortality without early treatment. Early treatment and prophylaxis with streptomycin or gentamicin or the tetracycline or fluoroquinolone classes of antimicrobials would be advised. PMID- 10807390 TI - Child injuries and fatalities from alcohol-related motor vehicle crashes: call for a zero-tolerance policy. PMID- 10807391 TI - Avalanches, air pockets, and advertisements. PMID- 10807392 TI - The new Atlantis and the frontiers of medicine. PMID- 10807393 TI - Optical prosthesis: visions of the future. PMID- 10807394 TI - Examining the living genome in health and disease with DNA microarrays. PMID- 10807395 TI - Neural stem cells and gene therapy: prospects for repairing the injured spinal cord. PMID- 10807396 TI - JAMA Patient Page: alcohol and driving. PMID- 10807397 TI - Review article: chronic inflammation and reactive oxygen and nitrogen metabolism- implications in DNA damage and mutagenesis. AB - It is well known that chronic inflammation of the digestive tract is associated with an increased risk of malignant transformation. Because phagocytic leukocytes and cytokine-activated parenchymal cells produce large amounts of reactive metabolites of oxygen and nitrogen, there has been substantial interest in ascertaining whether these reactive intermediates may mediate mutagenesis and malignant transformation in vivo. However, very little information is available regarding the basic chemistry of how these oxygen and nitrogen-derived species may interact to yield potentially carcinogenic agents. This review will discuss our present understanding of the chemical and biochemical interactions between superoxide and nitric oxide and provide a model by which these reactive species may damage DNA and mediate mutagenesis. PMID- 10807398 TI - The effects of aspirin on antioxidant defences of cultured rat gastric mucosal cells. AB - BACKGROUND: Helicobacter pylori-associated inflammation leads to exposure of the gastric epithelium to reactive oxygen species (ROS) generated in the gastric mucosa. In some pathological conditions, such as those induced by nonsteroidal anti-inflammatory drugs, the gastric mucosa may become more susceptible to ROS. AIM: To examine the effects of aspirin on antioxidant defenses as well as on oxidant injury in cultured rat gastric mucosal cells. METHODS: Primary monolayer cultures of rat gastric fundic mucosa were exposed to an ROS-generating system, hypoxanthine/xanthine oxidase (XOD). Cytotoxicity was quantified by measuring 51Cr release from prelabelled cells. The effects of aspirin on antioxidants and on cellular injury brought about by the ROS-generating system were determined. RESULTS: XOD, in the presence of hypoxanthine, caused a dose-dependent increase in specific 51Cr release, which corresponded to the ability of XOD to produce ROS (as assessed by the production of uric acid from hypoxanthine). Incubation of cells with aspirin (1-100 microM) produced a dose-dependent increase in XOD induced 51Cr release. Aspirin did not affect cellular glutathione content or activity of glutathione peroxidase, glutathione reductase or endogenous catalase. By contrast, aspirin caused a dose-dependent reduction in mucus synthesis. as assessed by incorporation of [3H]-glucosamine hydrochloride into the cells. CONCLUSIONS: Aspirin at therapeutically relevant concentrations rendered cultured gastric cells more susceptible to subsequent exposure to ROS. Aspirin affected neither the glutathione redox cycle nor catalase activity. Thus, the enhancement of ROS-induced injury by aspirin may be accomplished through diminished gastric mucus synthesis, since mucus is a potent scavenger of ROS. These findings provide insight into how gastric inflammation and injury (such as that induced by H. pylori infection) in human gastric mucosa is modulated by the administration of nonsteroidal anti-inflammatory drugs. PMID- 10807399 TI - The roles of nitric oxide and prostaglandins in alterations of ulcerogenic and healing responses in adjuvant-induced arthritic rat stomachs. AB - AIM: To examine alterations of gastric ulcerogenic and healing responses in adjuvant-induced arthritic rats. METHODS: Arthritis was induced in male Dark Agouti rats by injection of Freund's complete adjuvant into the right hind paw. RESULTS: The gastric ulcerogenic response to indomethacin was markedly increased in AA rats, depending on the degree of arthritic change. By contrast, HCl/ethanol induced gastric lesions were significantly suppressed in arthritic rats when compared with normal rats. The increased ulcerogenic response to indomethacin was significantly prevented by L-NAME and antineutrophil serum but not by FR167653, while the reduced ulcerogenic response to HCl/ethanol was significantly prevented by L-NAME and partly by indomethacin or NS-398. On the other hand, the healing of chronic gastric ulcers induced by thermal cauterization was also significantly delayed in arthritic rats when compared with normal rats. This delayed healing of gastric ulcers was affected by neither L-NAME, indomethacin nor FR167653. The gastric mucosa of arthritic rats showed a significant increase in both inducible nitric oxide synthase (iNOS) activity and prostaglandin (PG) E2 contents. CONCLUSIONS: Gastric ulcerogenic and healing responses were altered in arthritic rats. The ulcerogenic response to indomethacin was increased while that to HCl/ethanol was decreased. These changes in ulcerogenic responses may both be accounted for by increased production of NO/iNOS, with the latter also being partially related to elevated production of PGs/COX-2. Moreover, the healing of gastric ulcers was also delayed in arthritic rats, but the mechanism was related to neither NO nor PGs. PMID- 10807400 TI - Role of inducible nitric oxide synthase in dextran sulphate sodium-induced colitis. AB - BACKGROUND: Different authors have postulated both toxic and protective effects for nitric oxide (NO) in the pathophysiology of active inflammation. AIM: To examine the role of NO, especially that produced by the inducible form of nitric oxide synthase (iNOS), by investigating the effects of NOS inhibitors and NO donors on inflammation in experimental acute colitis. METHODS: Acute colitis was induced in rats by dextran sulphate sodium (DSS). White blood cell counts and levels of thiobarbituric acid reactants in the portal blood were determined, as were histological changes in the colonic mucosa. We then evaluated the effects of N(G)-nitro-L-arginine methyl ester (L-NAME), aminoguanidine (AG) and an NO donor on DSS-induced changes in these inflammatory parameters. RESULTS AND CONCLUSIONS: Inhibition of NO production by either L-NAME or AG worsened DSS-induced inflammation, suggesting a protective role for NO in acute colitis. On the other hand, a NO donor also exaggerated DSS-induced inflammatory parameters, suggesting that acute colitis may be aggravated by either too much or too little NO. These results suggest that medical treatment of ulcerative colitis must aim for maintenance of appropriate NO levels in the intestinal mucosa. PMID- 10807401 TI - Review article: transcription factors and growth factors in ulcer healing. AB - This review is focused on recent investigations demonstrating a pharmacological and pathophysiologic role in gastroduodenal ulceration for growth factors such as basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF), as well as for transcription factors. Our experiments revealed accelerated healing, without decreased gastric acid secretion, of chronic cysteamine-induced duodenal ulcers in rats treated daily for 3 weeks with intragastric administration of bFGF, PDGF or VEGF. Our recent studies also indicate a pathophysiological role of endogenous growth factors in the natural history of experimental duodenal ulcer development and healing. More recently, we investigated the genetic regulation of these growth factors in experimental duodenal ulceration. Since gene expression is most effectively controlled by transcription factors, proteins that bind to cis-acting elements of DNA and guide the binding of polymerase II to start the transcription of specific mRNA, we tested the hypothesis that the expression of IEGs and their transcription factor products, such as Egr-1 and Sp1, might precede the increased synthesis of bFGF, PDGF and VEGF in duodenal ulcer healing. Indeed, the duodenal ulcerogen cysteamine, but not its nonulcerogen and toxic analogue ethanolamine, rapidly increased duodenal (but not gastric) mucosal levels of ET-1, which was followed by enhanced expression of Egr-1 and a decrease in Sp1 in the preulcerogenic stage of duodenal ulceration. These changes in levels of ET-1 and expression of transcription factors were also accompanied by increased expression of the CDK inhibitor p21. Thus, not only growth factors such as bFGF, PDGF and VEGF, but also transcription factors such as Egr-1 and Sp1 and the cell cycle regulator p21, may play a role in the natural history of experimental duodenal ulceration. PMID- 10807402 TI - The effect of NSAIDs and a COX-2 specific inhibitor on Helicobacter pylori induced PGE2 and HGF in human gastric fibroblasts. AB - BACKGROUND: There is compelling evidence for the pivotal role of Helicobacter pylori in the pathogenesis of gastrointestinal ulcer disease. However, despite the bacterium's toxicity, the majority of H. pylori infections are not accompanied by gastric ulcers. This implies the existence of a host mechanism offsetting H. pylori toxicity. AIMS: To evaluate gastric fibroblasts' expression of hepatocyte growth factor (HGF), which is known to facilitate gastric ulcer healing, in the presence of H. pylori; to compare the effect on H. pylori-induced HGF expression of a COX-2 selective inhibitor with that of nonselective nonsteroidal anti-inflammatory drugs METHODS: Human gastric fibroblasts were cultured from human gastric mucosa obtained at surgery. Prostaglandin E2 (PGE2) and HGF were measured by EIA. The expression of COX-2 mRNA was assessed by the TaqMan quantitative RT-PCR system. RESULTS: H. pylori increased PGE2 release in gastric fibroblasts. H. pylori induced expression of COX-2 mRNA, which indicates that PG induction by H. pylori is through COX-2. Sulindac sulphide, etodolac and NS 398 all inhibited H. pylori-induced PGE2 release to the same extent. These agents also inhibited H. pylori-induced HGF release. CONCLUSION: Gastric fibroblasts produce PG and HGF in response to the presence of H. pylori, which may be considered part of the human body's defensive reaction to H. pylori toxicity. This defensive mechanism is inhibited not only by COX-2 nonselective NSAIDs but also by a COX-2 selective inhibitor. These findings indicate the importance of COX-2 in chronic H. pylori infection. PMID- 10807403 TI - Autonomic nervous regeneration in acetic acid-induced ulcer from the viewpoint of synapse formation--effect of basic fibroblast growth factor and sofalcone in the rat. AB - BACKGROUND: Monoclonal antibodies against GAP43 and synaptophysin, markers of regenerated nerves, have recently become available. AIM: To investigate the regeneration of the autonomic nerves after acetic acid treatment, as well as the effect of recombinant basic fibroblast growth factor (bFGF-CS23) and sofalcone on reinnervation. METHODS: Ulcers were induced by the direct application of 100% acetic acid to the serosal surface of the rat fundic stomach. Some rats were treated with bFGF-CS23 or sofalcone every 12 h after the acetic acid treatment. The immunohistochemical location of GAP43 and synaptophysin was observed by confocal laser microscopy, and the uptake sites of 14C-sofalcone were observed by autoradiography. RESULTS: Both GAP43 and synaptophysin immunoreactivities surrounding microvessels were weak in the control group, whereas in the acetic acid-treated group, these immunoreactivities were increased. Treatment with bFGF CS23 and sofalcone increased these immunoreactivities. The binding sites of sofalcone coincided with the location of regenerated nerves and surface mucous cells. The progenitors of the autonomic nerves were more abundant than expected. CONCLUSION: Both bFGF and sofalcone seem to stimulate nerve regeneration. PMID- 10807404 TI - Induction of cyclooxygenase-2 in mesothelial cells in peritonitis caused by perforated ulcers--an immunohistochemical study in humans. AB - BACKGROUND: Increasing evidence suggests that mesothelial cells contribute to the control of inflammation in the peritoneal cavity by secreting prostaglandins. A study has shown that cyclooxygenase (COX)-2 knockout mice die partly as a result of peritonitis. AIM: To investigate the expression and location of COX in peritonitis associated with peptic ulcer perforation. METHODS: Gastric and duodenal tissues were collected intraoperatively from nine and four patients, respectively, and immunohistochemical staining for COX-1 and COX-2 was performed. RESULTS: Histologically, all patients had severe peritonitis around the perforation sites, into which many inflammatory cells and fibroblasts had infiltrated, and reactive mesothelial cells exhibited hyperplastic change. The COX-1 protein was not detected, whereas COX-2 was abundant in reactive mesothelial cells near the perforation site and disappeared away from the site. Macrophages and fibroblasts around the perforation site also revealed immunostaining for COX-2. CONCLUSIONS: Our results showed that COX-2 protein is induced in mesothelial cells, as well as in macrophages and fibroblasts, in inflamed peritoneal tissues associated with peptic ulcer perforation, suggesting involvement of COX-2 in tissue repair. PMID- 10807405 TI - Review article: cyclooxygenase--a target for colon cancer prevention. AB - Use of nonsteroidal anti-inflammatory drugs such as aspirin, which are known to inhibit cyclooxygenase activity, reduces the relative risk of colorectal cancer in humans by 40-50%. Animal and human studies have shown a 50-80% reduction in tumour multiplicity following treatment with a variety of nonsteroidal anti inflammatory drugs. Two isoforms of cyclooxygenase have been described, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). In 85% of colorectal adenocarcinomas taken from humans. COX-2 levels are 2-50-fold higher than levels in adjacent normal intestinal mucosa, while COX-1 levels are unchanged. These observations raise the question: Does COX-1 or COX-2 provide a useful target for prevention or treatment of colorectal cancer? PMID- 10807406 TI - Gastric epithelial cell proliferation and apoptosis in Helicobacter pylori infected mice. AB - BACKGROUND: Helicobacter pylori causes gastritis and is strongly associated with gastroduodenal ulcer and gastric cancer. The bacterium is associated with an increased rate of epithelial proliferation, which can be reversed by eradication of the organism. The mechanism of this response is not known, but this epithelial proliferation is one of the risk factors for developing gastric carcinoma. Recently, apoptosis also was found to be increased in the gastric mucosa of persons carrying H. pylori. METHODS: cagA-positive H. pylori isolated from a human gastric ulcer was inoculated into BALB/C mice. At 4, 6, 12, 18 and 24 weeks, mice were injected with bromodeoxyuridine 5 mg/kg and killed 1 h later. Proliferation was analysed by histochemical staining for BrdU; apoptosis was examined by the TUNEL assay. RESULTS: The number of BrdU-labelled cells in the antrum was significantly increased by H. pylori infection beginning 12 weeks after infection. The number of apoptotic cells in this tissue was increased significantly by 6 weeks after inoculation. CONCLUSION: The proliferation observed in H. pylori infection may be a response to increased apoptosis. PMID- 10807407 TI - A new mechanism for anti-inflammatory actions of proton pump inhibitors- inhibitory effects on neutrophil-endothelial cell interactions. AB - BACKGROUND: Neutrophil-endothelial cell interactions mediated by adhesion molecules may be involved in gastric mucosal inflammation associated with Helicobacter pylori or nonsteroidal anti-inflammatory drugs. AIM: To investigate the effects of proton pump inhibitors and histamine-2 receptor antagonists (HRA) on neutrophil-endothelial cell adhesive interactions induced by H. pylori water extract (HPE) or interleukin-1beta (IL-1beta). METHODS: Human peripheral neutrophils and umbilical vein endothelial cells were incubated with either proton pump inhibitors (lansoprazole and omeprazole) or HRA (famotidine and ranitidine). Neutrophil surface expression of CD11b and CD18 and endothelial cell intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM 1) were assessed by flow cytometry and an enzyme immunoassay, respectively. Neutrophil adherence was defined as the ratio of exogenous neutrophils that adhered to the endothelial monolayers. RESULTS: The expression of CD11b and CD18 on neutrophils and neutrophil-dependent adhesion to endothelial cells elicited by HPE were inhibited by lansoprazole and omeprazole at clinical relevant doses, and the expression of ICAM-1 and VCAM-1 on endothelial cells and endothelial dependent neutrophil adherence induced by IL-1beta were also inhibited by lansoprazole and omeprazole at similar doses. Famotidine and ranitidine had no effect on neutrophil-endothelial cell interactions. CONCLUSIONS: These results indicate that proton pump inhibitors can attenuate neutrophil adherence to endothelial cells via inhibiting the expression of adhesion molecules, suggesting that proton pump inhibitors may have anti-inflammatory activity. PMID- 10807408 TI - Is autonomic dysfunction a necessary condition for chronic peptic ulcer formation? AB - BACKGROUND: The relationship between 1/f fluctuation of the heart rate variability and Helicobacter pylori infection was evaluated, in order to clarify whether autonomic nervous dysfunction is a necessary condition for chronic peptic ulcer formation. METHODS: The subjects were 11 patients with recurrent chronic peptic ulcer and 20 age-matched normal subjects. Holter ECGs were recorded over 24 h, and the 1/f(-x) fluctuation of the heart rate was computed. The 1/f(-x) fluctuation of the heart rate is a novel index of autonomic function that has been shown to reflect a patient's pleasant mood. For 1/f(-x) fluctuation, the slope of the regression line (-x) was determined and cosine fitting of the absolute slope of the regression line over a 24-h period was performed. RESULTS: For the normal group, the absolute slope of the regression line during daytime and night-time were 0.53 +/- 0.16 and 1.05 +/- 0.18, respectively. For the peptic ulcer group, the corresponding values during daytime and night-time were 0.94 +/- 0.15 and 1.84 +/- 0.18, respectively. The mean value of the cosine curve was 0.76 +/- 0.23 in the normal group and 1.12 +/- 0.25 in the peptic ulcer group. Thus, these values were significantly higher for the latter group than for the former group (P<0.05). No statistically significant difference in H. pylori infection between the two groups was observed. CONCLUSIONS: Autonomic nervous dysfunction as well as H. pylori infection appears to be a necessary condition for chronic peptic ulcer formation. PMID- 10807409 TI - Review article: healing after inflammatory injury--coordination of a regulatory peptide network. AB - Intestinal epithelial cells are capable of producing a variety of cytokines and other regulatory factors that can affect functional regulation of the epithelium itself, through autocrine and paracrine mechanisms, as well as functional integration with lamina propria populations. The bi-directional nature of this cytokine network is now apparent, with the demonstration that both rat and human intestinal epithelium-derived cell lines possess a much greater array of cytokine receptors than previously anticipated. PMID- 10807410 TI - Rat CXC chemokine GRO/CINC-1 paradoxically stimulates the growth of gastric epithelial cells. AB - BACKGROUND: CXC chemokines such as interleukin (IL)-8 are neutrophil chemoattractants, the levels of which increase in Helicobacter pylori-infected gastric mucosa. Many investigators have focused on the chemotactic aspects of IL 8: however, CXC chemokines are also reported to have angiogenic activity and to serve as remodelling factors. Rat GRO/CINC-1 is a rodent counterpart of human GROalpha, a member of the family of CXC chemokines. Gastric mucosa infected with H. pylori is in a state of hyperproliferation, with increases in the amounts of growth factors such as hepatocyte growth factor (HGF). AIM: To investigate whether rat GRO/CINC-1 had growth-stimulating activity for gastric epithelial cells. METHODS: The rat gastric epithelial cell line RGM-1 was incubated in serum free medium for 12 h to adjust the cell cycle to the G0 phase, and GRO/CINC-1 was then added for 24 h. The total cell number was determined by fluorogenic analysis after propidium iodide staining, and cell proliferation was assessed by measuring 5-bromo-2'-deoxyuridine (BrdU) incorporation. The activity of p42/p44 mitogen activated protein kinase (MAPK) was measured 5-20 min after the start of GRO/CINC 1 exposure. RESULTS: Cultures treated with GRO/CINC-1 showed a significant increase in cell number and BrdU incorporation in a concentration-dependent fashion. The MAPK activity increased within 5 min after GRO/CINC-1 application and returned to the control level at 20 min. CONCLUSION: The growth-stimulatory effect of GRO/CINC-1 on rat gastric epithelial cells suggests a dual function of this chemokine: proinflammatory action and induction of epithelial proliferation. PMID- 10807411 TI - Induction of signal transduction pathways in rat gastric epithelial cells stimulated with interleukin-1beta. AB - BACKGROUND: Interleukin-1beta (IL-1beta) participates in cell growth, differentiation and apoptosis via activation of several kinases in a variety of cells. Mitogen-activated protein (MAP) kinases are important intermediates of the signal transduction pathway from the cell surface to the nucleus, leading to activation of transcription factors. There are no reports on the effect of IL 1beta on these pathways in gastric epithelial cells. AIM: To investigate whether IL-1beta activates MAP kinases [extracellular signal-regulated kinases (ERKs), c Jun NH2-terminal kinases (JNKs) and p38 MAP kinase (p38 MAPK)] and nuclear factor (NF)-kappaB. a transcription factor, in gastric epithelial cells (RGM1). METHODS: The activities of ERKs and JNKs were estimated by in-gel kinase assay, and p38 MAPK activity was measured by in vitro kinase assay at various time points (0-40 min) after addition of IL-1beta (100 pg/mL) for 20 min. The activity of NF-kappaB was analysed using gel mobility shift assay at times from 0 to 4 h after addition of IL-1beta. RESULTS: Activity of ERKs was detectable at 10 min, peaked at 20 min, and continued at increased levels until 40 min. Activity of both JNKs and p38 MAPK were detectable during 5-20 min, and then decreased within 40 min. Activation of NF-kappaB occurred at 30 min, and increased activity continued for 6 h. Interleukin-1beta activated MAP kinases and NF-kappaB in RGM1 cells. CONCLUSION: The activation induced by this cytokine may play an important role in the initiation of the inflammatory process in gastric mucosa. PMID- 10807412 TI - Gastric acid secretion in dogs in response to combinations of beer, ethanol and peptone meal--the role of endogenous gastrin. AB - AIM: To examine the effects of beer, ethanol, peptone meal and combinations of either peptone meal and beer or peptone meal and ethanol on gastric acid secretion in vagally denervated pouch dogs. METHODS AND RESULTS: The oral administration of either 200 mL of beer, 5% ethanol or 10% peptone meal significantly stimulated gastric acid secretion for 60-90 min in these dogs. With 5% ethanol the plasma gastrin concentration was not affected for 90 min. Combinations of 10% peptone and beer (peptone-beer) or 10% peptone and 5% ethanol (peptone-ethanol) potentiated the acid secretion and increased the plasma gastrin level. While a selective cholecystokinin-B/gastrin receptor antagonist, S-0509, had no effect on ethanol-stimulated acid secretion, the compound markedly inhibited both peptone-beer-stimulated and peptone-ethanol-stimulated gastric acid secretion. Famotidine and atropine significantly inhibited gastric acid secretion stimulated by 5% ethanol, peptone-beer and peptone-ethanol. CONCLUSIONS: The mechanisms by which peptone-beer and peptone-ethanol stimulate gastric acid secretion may be mediated not only by increased plasma gastrin, but also by the action of histamine and acetylcholine coupled with the increased plasma gastrin. Ethanol-stimulated secretion appears to be unrelated to circulating gastrin, yet may be related to the acid regulatory mechanism involving histamine and acetylcholine. PMID- 10807413 TI - The roles of prostaglandin E receptor subtypes in the cytoprotective action of prostaglandin E2 in rat stomach. AB - AIM: To investigate the EP receptor subtype involved in the gastroprotective action of prostaglandin (PG) E2 using various EP receptor agonists in rats, and using knockout mice lacking EP1 or EP3 receptors. METHODS: Male SD rats and C57BL/6 mice were used after an 18-h fast. Gastric lesions were induced by oral administration of HCl/ethanol (150 mM HCl in 60% ethanol). Rats were given various EP agonists i.v. 10 min before HCl/ethanol: PGE2, sulprostone (EP1/EP3 agonist), butaprost (EP2 agonist), 17-phenyl-omega-trinorPGE2 (17-phenylPGE2: EP1 agonist), ONO-NT012 (EP3 agonist) and 11-deoxyPGE1 (EP3/EP4 agonist). In a separate study, the effect of PGE2 on HCl/ethanol lesions was examined in EP1- and EP3-receptor knockout mice. RESULTS: Gastric lesions induced by HCl/ethanol were dose dependently prevented by PGE2: this effect was mimicked by sulprostone and 17-phenylPGE2 and was significantly antagonized by ONO-AE-829, an EP1 antagonist. Neither butaprost, ONO-NT012 nor 11-deoxyPGE1 exhibited any protective activity against HCl/ethanol-induced gastric lesions. PGE2 caused an inhibition of gastric motility as well as an increase of mucosal blood flow and mucus secretion, the effects being mimicked by prostanoids activating EP1 receptors, EP2/EP3/EP4 receptors and EP4 receptors, respectively. On the other hand, although HCl/ethanol caused similar damage in both wild-type mice and knockout mice lacking EP1 or EP3 receptors, the cytoprotective action of PGE2 observed in wild-type and EP3-receptor knockout mice totally disappeared in mice lacking EP1 receptors. CONCLUSION: The gastric cytoprotective action of PGE2 is mediated by activation of EP1 receptors. This effect may be functionally associated with inhibition of gastric motility but not with increased mucosal blood flow or mucus secretion. PMID- 10807414 TI - Interactive roles of endogenous prostaglandin and nitric oxide in regulation of acid secretion by damaged rat stomachs. AB - BACKGROUND: The acid inhibitory mechanism in the damaged stomach is known to involve endogenous nitric oxide (NO) as well as prostaglandin (PG). AIM: To investigate the interaction between PG and NO in regulation of acid secretion in the stomach following damage. METHODS: Under urethane anaesthesia, a rat stomach was mounted in an ex vivo chamber and perfused with saline. Acid secretion, luminal PGE2, NO metabolites (NOx) and histamine output were measured before and after application of 20 mM taurocholate Na (TC) for 30 min, with or without pre treatment with indomethacin and/or N(G)-nitro-L-arginine methyl ester (L-NAME). RESULTS: Exposure of the stomach to TC caused a decrease in acid secretion, with concomitant increase of both luminal NOx and PGE2. Either L-NAME or indomethacin reduced the decrease in acid secretion in response to TC, but only L-NAME allowed acid secretion to increase over basal values. L-NAME prevented the increase of luminal NOx after TC treatment, while indomethacin inhibited PGE2 release during and after exposure to TC. The increase in acid secretion in the presence of L NAME was prevented when indomethacin was given concomitantly. TC treatment increased histamine output in the lumen, a process that was enhanced by L-NAME but reduced by indomethacin. CONCLUSIONS: Damage to the stomach increases both NO and PG in the lumen, and decreases acid secretion. Inhibiting NO production increases acid secretion in the damaged stomach, but only when PG biosynthesis is intact. It is assumed that endogenous PG has a dual role in the regulation of acid secretion in the damaged stomach: an inhibitory effect at the parietal cell and an excitatory effect probably through enhancing the release of mucosal histamine. PMID- 10807415 TI - Mechanism of prevention by capsaicin of ethanol-induced gastric mucosal injury--a study in the rat using intravital microscopy. AB - BACKGROUND: Capsaicin acts specifically on primary afferent neurones to release neuropeptides, including calcitonin gene-related peptide (CGRP), and prevents ethanol-induced mucosal injury. AIM: To investigate the microvascular changes in the gastric mucosa in response to ethanol using intravital microscopy to elucidate the mechanism of capsaicin-induced gastroprotection. METHODS: The posterior gastric wall in the rat was secured in an observation chamber and perfused with Tyrode's solution. The microcirculation was observed through a window made by removing a limited area of smooth muscle. RESULTS: Ethanol (50%) applied to the mucosa constricted the collecting venules and venules but dilated arterioles. The constriction of the collecting venules resulted in mucosal congestion, which caused mucosal injury. Application of capsaicin to the mucosa dilated the arterioles but not the collecting venules or venules. Arteriolar dilation was inhibited by a CGRP antagonist, CGRP-(8-37). Prior application of capsaicin prevented ethanol-induced constriction of the collecting venules, and the action of capsaicin was inhibited by prior application of CGRP-(8-37). CONCLUSIONS: The results suggest that the inhibition of ethanol-induced gastric injury by capsaicin is attributable to the suppression of collecting venule constriction, via CGRP release. PMID- 10807416 TI - Protective role of intracellular glutathione against nitric oxide-induced necrosis in rat gastric mucosal cells. AB - BACKGROUND: Nitric oxide synthase activity is increased in the stomach in association with Helicobacter pylori infection and portal hypertension, but the mechanism by which nitric oxide contributes to mucosal damage remains unclear. AIM: To examine whether nitric oxide injures gastric mucosal cells and whether cellular glutathione affects nitric oxide-induced cytotoxicity. METHODS: A confluent monolayer of RGM-1 gastric mucosal cells was exposed to nitric oxide donors (NOC5 or NOC12). Cell viability was determined by trypan blue dye exclusion, lactate dehydrogenase release and supravital staining with Hoechst 33342 and propidium iodide. The kinetics of the reduced/oxidized forms of glutathione were also measured, as well as the effect of glutathione-depletion or glutathione-precursor treatment on nitric oxide-induced cytotoxicity. RESULTS: Excess exogenous nitric oxide produced by NOC5 or NOC12 induced necrosis in RGM-1 cells in a time- and concentration-dependent manner. The level of reduced glutathione drastically decreased prior to the loss of cell viability and remained low, but oxidized glutathione was not affected. Glutathione depletion increased necrosis of both NOCs in an NOC-concentration-related fashion, while pre-treatment with gamma-glutamylcysteine ethyl ester reduced their necrotic susceptibility. CONCLUSION: Exogenous nitric oxide induced necrosis in gastric mucosal cells, and intracellular reduced glutathione protects gastric mucosal cells from damage by nitric oxide. PMID- 10807417 TI - Review article: the stages of gastrointestinal carcinogenesis--application of rodent models to human disease. AB - The development of gastrointestinal cancer in humans and animals occurs through a consecutive series of stages termed initiation, promotion and progression. The characterization of each of these stages has been elucidated in several model systems as well as in human neoplasms. Both single, putatively initiated cells and preneoplastic foci have been identified by marker protein differences as well as by mutational changes. The promotion stage involves the clonal expansion of single initiated cells. Such expansion can be rapidly reversed by a variety of means, of which acute fasting (as exemplified in rat hepatocarcinogenesis) is among the most rapid and efficient. This reversal involves a selective apoptosis of preneoplastic cells and preneoplastic lesions, associated with a marked increase in expression of the proto-oncogene c-myc. Transition of cells from the stage of promotion to that of progression initially involves specific karyotypic alterations, as noted in both the rat liver model and human colon carcinogenesis. In the former, the transition appears to be associated with enhanced expression of the H119 imprinted putative tumour suppressor gene. Thus, the use of model systems may be applied directly to the human circumstance, increasing the potential both for rational prevention of gastrointestinal neoplasia and for new approaches to the therapy of neoplastic disease in the progression stage. PMID- 10807418 TI - The incidence of reflux oesophagitis after cure of Helicobacter pylori in a Japanese population. AB - AIM: To investigate the incidence of reflux oesophagitis after antibacterial therapy for Helicobacter pylori infection in our patient population. METHODS: Subjects were 451 H. pylori-infected patients (primary symptom: peptic ulcer disease in 347, nonulcer dyspepsia in 100, and reflux oesophagitis in four): 11 of these patients had reflux oesophagitis on study entry. H. pylori infection was treated by a proton pump inhibitor/amoxycillin-clarithromycin regimen for either 7 or 14 days. Each patient was examined by endoscopy before treatment and more than 6 months after treatment to compare oesophageal findings. In addition, 227 patients were interviewed regarding reflux symptoms, using symptom questionnaires, before and more than 6 months after treatment. RESULTS: Among 440 patients who did not have reflux oesophagitis prior to antibacterial treatment (340 peptic ulcer patients and 100 nonulcer dyspepsia patients), 23 patients whose infection was eradicated developed reflux oesophagitis (5.4%). The 11 patients who had reflux oesophagitis prior to treatment were all successfully cured of infection. Six of these patients showed no change in their oesophagitis, while the condition improved in three and worsened in two. Symptom scores improved in 34 of the 36 patients who reported reflux symptoms. Among 19 patients who showed persistent infection, only one developed reflux oesophagitis (5.2%), while none complained of newly developed symptoms following treatment. CONCLUSIONS: Development of reflux oesophagitis after treatment of H. pylori infection was observed in a Japanese population. However, the incidence of this condition was comparable between those with persistent H. pylori infection and those in whom the infection was eradicated. PMID- 10807419 TI - Manometric and hormonal changes after distal partial gastrectomy. AB - BACKGROUND: Alkaline oesophagitis attributable to duodenal mechanisms may induce oesophageal carcinogenesis in a rat reflux model. AIM: To investigate the mechanism of the regurgitation after distal partial gastrectomy. METHODS: Oesophageal manometry was used in 16 patients before and after distal partial gastrectomy with reconstruction by Bilroth methods. Serum concentrations of four gastrointestinal hormones were measured by radioimmunoassay in 10 gastrectomy patients and nine healthy volunteers before and after a standardized meal. RESULTS: The lower oesophageal sphincter pressure was reduced to 83% after surgery. The amplitude and duration of the peristaltic waves tended to be increased, and the velocity tended to be less after surgery (amplitude 120%, duration 114%, velocity 88%). Interrupted waves appeared more frequently after surgery. The manometric changes in gastrectomized patients are considered to be disadvantageous relative to regurgitation. After surgery, gastrin and pancreatic polypeptide were completely abolished postprandially, whereas cholecystokinin and neurotensin were significantly increased. CONCLUSION: The hormonal changes should have a suppressive effect on the lower oesophageal sphincter. Both the manometric and the hormonal changes may exacerbate reflux oesophagitis after distal partial gastrectomy. PMID- 10807420 TI - Review article: transcriptional events controlling the terminal differentiation of intestinal endocrine cells. AB - Secretin-producing enteroendocrine cells arise from a multipotential endocrine progenitor in the crypts of the small intestine. As these cells migrate up the crypt-villus axis, they produce secretin and stop dividing as they terminally differentiate and die. Transcription of the secretin gene is controlled by a complex enhancer binding to multiple transcription factors. The basic helix-loop helix protein, BETA2, binds to an E box sequence and associates with the p300 coactivator to activate transcription of the secretin gene. Basic helix-loop helix proteins appear to play a pivotal role in the control of cellular differentiation. BETA2 induces cell cycle arrest and apoptosis in addition to activating secretin gene expression. Thus BETA2 may function as a master regulatory gene to coordinate terminal differentiation of secretin cells. PMID- 10807421 TI - Effects of growth factors on aspirin-induced inhibition of wound repair in a rabbit gastric epithelial cell model. AB - BACKGROUND: Aspirin is known to cause adverse effects, including gastric mucosal injury, and to retard gastric wound healing. Growth factors including hepatocyte growth factor (HGF), epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) have been shown to play an important role in the repair of gastric mucosal injury. AIM: To employ the cultured gastric epithelial cell model to elucidate the effects of aspirin, as well as several growth factors (HGF, EGF and IGF-I), on gastric wound repair. METHODS: Isolated rabbit gastric epithelial cells (92% mucous cells) were cultured in F-12 medium and formed a complete monolayer cell sheet in 48 h. A wound with a cell-free area of constant size (2 mm2) was then created and the wound repair process was monitored by measuring wound size every 12 h. Proliferating cells were detected by BrdU staining. Effects of aspirin (8 mM), HGF (10 ng/mL), EGF (10 ng/mL) and IGF-I (30 ng/mL) were assessed. RESULTS: Aspirin significantly retarded wound healing, but simultaneous addition of growth factors significantly accelerated wound repair compared with aspirin alone. Growth factors reversed the aspirin-induced inhibition of cell proliferation. CONCLUSION: Growth factors, including HGF, EGF and IGF-I, reversed the aspirin-induced inhibition of wound repair through their cytoprotective effects on gastric epithelial cells. PMID- 10807422 TI - Effect of activin on cell growth in primary cultures of guinea pig gastric epithelial cells. AB - BACKGROUND: The effect of activin on differentiated cells is known to be different from that on undifferentiated cells. Cultured gastric epithelial cells in complete serum-free conditions grew into matured mucous cells after treatment with epidermal growth factor (EGF). AIM: To elucidate the effect of activin on the growth of differentiated and undifferentiated gastric mucosal cells. METHODS: Cultured guinea pig gastric epithelial cells were prepared using the method of Ogihara et al. Synthesis of activin was analysed by Western blot using monoclonal anti-activin A antibody. Cell proliferation was assessed by counting the number of cells. Mucin production was assessed by histochemical study using periodic acid-Schiff (PAS) reaction. RESULTS: Western blot analysis indicated that activin was synthesized in cultured guinea pig gastric mucosal cells. One hundred nanomolar EGF induced a 3-fold increase in cell count and the appearance of PAS positive granules. Five nanograms activin per millilitre without EGF stimulated proliferation of the cells that showed almost negative PAS staining. When activin was added after treatment with 100 nM EGF for 24 h, cell proliferation induced by EGF was inhibited by activin at concentrations higher than 5 ng/mL. CONCLUSION: These results suggest that activin stimulates proliferation of undifferentiated cells and inhibits growth of differentiated cells. PMID- 10807423 TI - The effect of Helicobacter pylori on cell proliferation and apoptosis in gastric epithelial cell lines. AB - BACKGROUND: Helicobacter pylori has been implicated in the pathogenesis of gastric cancer and malignant lymphoma. It is not known whether the bacterium stimulates cell proliferation directly or if apoptosis induced by H. pylori leads to a hyperproliferative response. AIM: To clarify the precise mechanism of H. pylori action on gastric epithelial cell growth, we compared the response of two cell lines, Kato III (p53 deletion) and MKN 45 (p53 wild type), to the organism. To determine the role of Helicobacter vacuolating cytotoxin in gastric mucosal injury, we examined the relation between vacuolating activity and apoptosis under several conditions. METHODS: Five cytotoxic and four noncytotoxic strains of H. pylori were used, each with an inoculum of 10(7) cfu/mL. The effect on the growth in MKN 45 and Kato III cells was studied by MTT assay. Vacuolating cytotoxin activity was determined using RK-13 cells. RESULTS: Neither cytotoxic nor noncytotoxic strains induced apoptosis, but death of MKN 45 cells was induced by pre-treatment with interferon-gamma and culture with TNF-alpha. In contrast, some strains of H. pylori increased proliferation of Kato III cells. Furthermore, cell death induced by cytotoxic strains, but not noncytotoxic strains, was significantly augmented by amoxycillin 5-50 g/mL (P=0.0016). On the other hand, acid-treated supernatant fluids from cultures of H. pylori showed enhanced vacuolating activity but did not induce cell death, suggesting that death is attributable to some factor other than the cytotoxin. CONCLUSION: These findings suggest that H. pylori induces apoptosis by a means independent of vacuolating cytotoxin. PMID- 10807424 TI - Matrix metalloproteinase expression in cultured human gastric wall fibroblasts- interactions with Helicobacter pylori isolated from patients with ulcers. AB - BACKGROUND: Matrix metalloproteinases (MMPs), enzymes capable of degrading collagens and other extracellular matrix components, have been implicated in gastric ulcer formation. However, the effect on MMP expression of Helicobacter pylori, also implicated in these lesions, has not been studied to our knowledge. AIM: To seek links between H. pylori and MMP expression likely to affect gastric ulcer formation. After fibroblasts from human gastric wall were cocultured with H. pylori. concentrations of MMP-1 and -2 in the medium were determined by enzyme linked immunosorbent assays. RESULTS: Whereas MMP-1 was not detected in media from fibroblasts or H. pylori culture alone, MMP-1 was detected in cocultures (P<0.01). Similar amounts of MMP-2 were detected in medium from fibroblasts cultured alone and with H. pylori. No MMP-2 production by H. pylori cultured alone was detected. CONCLUSIONS: MMP-1 appears to be important in gastric ulcer pathogenesis, and MMP-1 induction by H. pylori may impede gastric ulcer healing. PMID- 10807425 TI - The effects of water extracts of CagA positive or negative Helicobacter pylori on proliferation, apoptosis and connexin formation in acetic acid-induced gastric ulcer of rats. AB - AIM: To investigate the influence of water extracts of CagA-positive or -negative Helicobacter pylori on healing of chronic gastric ulcers and on connexin formation, proliferation and apoptosis, in acetic acid-induced chronic gastric ulcers in rats. METHODS: Acetic acid was used to induce chronic gastric ulcers in rats, and a water extract of H. pylori was given by mouth every day. Connexin 32 formation was assessed using Western blotting as previously described. The frequencies of proliferating cell nuclear antigen staining and of TdT-mediated dUTP-biotin nick end labelling were examined. RESULTS: In untreated rats, acetic acid-induced gastric ulcers healed after 14 days and the electrophoretic band corresponding to connexin 32 appeared 4 days after ulcer induction. Treatment with a water extract of H. pylori delayed ulcer healing, with the ulcers remaining unhealed even on the 14th day; healing was delayed more when treatment was with an extract of CagA-positive rather than CagA-negative H. pylori. Connexin 32 appeared earlier when treatment was with a CagA-negative rather than a CagA-positive extract, but in both cases later than in the untreated control group. Proliferating cell nuclear antigen labelling on the fourth day was seen in 14.5% +/- 1.6% of mucosal cells of control group, but in 35.9% +/- 1.4% and 36.5% +/- 1.4% of mucosal cells treated with either VacA(+)CagA(-) and VacA(+)CagA(+) H. pylori extract, respectively. Furthermore, extracts of both H. pylori strains, especially VacA(+)Cag(+) H. pylori promoted apoptosis. CONCLUSIONS: A water extract of H. pylori increased both proliferation and apoptosis, which are related to exacerbation and healing of ulcer as well as appearance of connexin 32. PMID- 10807426 TI - Comparison between in vivo and in vitro chemokine production in Helicobacter pylori infection. AB - BACKGROUND: Enhanced gastric mucosal chemokine activity has been demonstrated in patients with Helicobacter pylori infection. However, little is known about the mechanisms involved in this phenomenon. AIM: To examine whether in vivo chemokine activity is similar to in vitro response of gastric epithelial cells infected by H. pylori. PATIENTS AND METHODS: Antral biopsy specimens were obtained from patients with H. pylori infection for organ culture, isolation of H. pylori and histological examination. RESULTS: In organ cultures of mucosal tissues, the levels of interleukin-8 and growth-related gene product a were elevated in patients with peptic ulcer disease compared with those with erosive gastritis or endoscopically normal mucosa. However, there were no significant differences in in vitro cultures of MKN45 or KATO III cells that were infected with H. pylori isolated from these same patients. These in vivo and in vitro alpha-chemokine levels showed no significant association with the presence of cagA gene and CagA protein, ureB genotype, or binding capacity to MKN45 or KATO III cells in individual H. pylori isolates. In contrast, in vivo mucosal alpha-chemokine activity correlated with H. pylori colonization density. CONCLUSION: Mucosal chemokine profiles and inflammatory responses in H. pylori infection may be associated more closely with host factors, including those determining bacterial adhesiveness, than with differences in H. pylori strains. PMID- 10807427 TI - Pharmacokinetics of clarithromycin in Helicobacter pylori eradication therapy in patients with liver cirrhosis. AB - BACKGROUND: Proton pump inhibitor triple therapy with clarithromycin and metronidazole has been widely used for Helicobacter pylori eradication. However, the efficacy and the safety of this therapy in patients with liver cirrhosis have not been established. AIM: To evaluate the effect of hepatic dysfunction on metabolism of clarithromycin as it is used for H. pylori eradication therapy in patients with liver cirrhosis, and the efficacy of eradication therapy in those patients. METHODS: Serum levels of clarithromycin and its metabolite. 14-(R) hydroxyclarithromycin, were examined in 18 subjects (five normal controls and 13 hospitalized patients with liver cirrhosis) on a selected day between days 7 and 10 of a 2-week course of eradication therapy. This therapy consisted of lansoprazole (30 mg, once a day) together with clarithromycin (200 mg, twice a day) and metronidazole (250 mg, twice a day). In addition, 118 H. pylori-positive out-patients, 88 with peptic ulcer and 30 with liver cirrhosis, underwent the same eradication therapy. RESULTS: Values for the area under the 0-6 h concentration-time curve (AUC) for clarithromycin were not significantly different among the groups. However, the AUC (0-6 h) values of 14-(R) hydroxyclarithromycin were significantly lower in the Child-Pugh C group than in either the normal controls or the Child-Pugh A/B group. The cure rate for the peptic ulcer patients was 84% on a per protocol analysis (95% CI: 80%-88%) and 81% on an intention-to-treat analysis (95% CI: 77%-85%), while in the liver cirrhosis patients it was 89% in a per protocol analysis (95% CI: 78%-99%) and 83% in an intention-to-treat analysis (95% CI: 70%-97%). Mild adverse effects were observed in 10% of the peptic ulcer patients and 13% of the liver cirrhosis patients, with none leading to premature withdrawal from the study. CONCLUSION: The 2-week low-dose lansoprazole-based triple therapy tested is a simple, effective and well-tolerated regimen for H. pylori eradication in patients with liver cirrhosis. PMID- 10807428 TI - The effect of sofalcone on indomethacin-induced gastric ulcers in a Helicobacter pylori-infected gnotobiotic murine model. AB - BACKGROUND: Sofalcone has been reported to exert anti-ulcer and gastroprotective actions, but its exact mechanism of action remains unknown. In our laboratory, we found that indomethacin-induced gastric ulcers become worse when associated with Helicobacter pylori infection. METHODS: We employed the H. pylori-infected gnotobiotic murine model to examine the effect of sofalcone on indomethacin induced gastric ulcers in the presence of H. pylori infection. In vitro experiments were also done to evaluate the effects of sofalcone on H. pylori growth, adherence of H. pylori to the MKN45 cells (a human gastric epithelial cell line), and these cells' IL-8 production in the presence of H. pylori. RESULTS: We found that sofalcone produced a significant improvement in ulcer size as well as a substantial reduction in the number of H. pylori colonies in H. pylori-infected gnotobiotic mice. In vitro sofalcone has a significant bacteriocidal effect against H. pylori and can also significantly prevent adherence of this bacterium to MKN45 cells, thus remarkably reducing IL-8 production of these cells in response to stimulation by H. pylori. CONCLUSION: Our results suggest that sofalcone can improve ulcer healing by the mechanisms mentioned above. PMID- 10807429 TI - A novel in vitro effect of the mucosal protective agent sofalcone--inhibition of chemotactic motility in Helicobacter pylori. AB - BACKGROUND: Motility of Helicobacter pylori is essential for colonization. H. pylori has been shown to exhibit chemotactic activity toward urea and sodium and bicarbonate ions, which are secreted from the gastric epithelia. The importance of urease activity for chemotactic motility of H. pylori in a viscous environment has also been shown. Consequently, application of drugs inhibiting chemotactic motility has been proposed as a strategy for H. pylori eradication. This inhibitory effect can be evaluated through assay of chemotaxis and swarming. MATERIALS AND METHODS: H. pylori CPY3401 and ATCC43504 were grown on brucella agar plates/broth supplemented with 3% horse serum under microaerobic conditions (N2, 85%; O2, 5%; CO2, 10%). For motility assay, H. pylori cells grown on brucella-serum agar were stabbed into motility agar containing 0.35% refined agar in brucella-serum broth and the swarming zone was measured. For the chemotaxis assay, cells were suspended in 10 mM potassium phosphate buffer, pH 7.0, with 3% polyvinyl-pyrrolidone and assayed as described previously. Bacterial swimming in the fluid environment was observed under dark-field microscopy. RESULTS: Numbers of bacteria attracted toward 1 microM flurofamide were reduced with increasing concentrations of sofalcone (0.2-222 microM). In addition, the size of the swarming zone was reduced in motility agar containing 22 and 222 microM sofalcone. On the other hand, 22 microM sofalcone did not inhibit bacterial growth on day 3. Bacterial swimming speed in brucella broth was slower in the presence of 22 and 222 microM sofalcone than in its absence. CONCLUSION: Sofalcone was found to inhibit chemotactic motility of H. pylori. This drug may be useful for inhibiting the bacterium's ability to colonize the human stomach. PMID- 10807430 TI - Surgery for cardiac myxomas. AB - Myxomas, particularly left-atrial myxomas, are the most common primary tumors of the heart that cardiac surgeons will be called upon to remove. Although some tumors are discovered incidentally during echocardiographic examination, many produce symptoms caused by the release of inflammatory cytokines, obstruction to intracardiac blood flow, and/or embolization. With rare exception, cardiac myxomas are benign, and excision is safe and curative in most patients. In a 38 year experience at the Mayo Clinic, 100 patients have had 106 operations for myxoma, and there has been only 1 perioperative death. During follow-up extending to 25 years, postoperative survival is similar to that of an age- and sex-matched population, and at 20 years postoperatively, 94% of patients were free of recurrent myxomas. Recognition of familial and syndrome myxomas (eg, Carney's complex) is important in guiding surgical approach, planning follow-up, and predicting recurrence of these unusual neoplasms. PMID- 10807431 TI - Unusual primary tumors of the heart. AB - Primary tumors of the heart, with the exception of atrial myxomas, occur rarely; tumors metastatic to or directly invasive of the heart are far more common. About 75% of primary tumors are benign, and 75% of these are atrial myxomas. The benign tumors include rhabdomyomas, fibromas, papillary fibroelastomas, hemangiomas, pericardial cysts, lipomas, hamartomas, teratomas, mesotheliomas, and paragangliomas or pheochromocytomas. The last 3 may also be malignant. The malignant tumors consist of various sarcomas: myxosarcoma, liposarcoma, angiosarcoma, fibrosarcoma, leiomyosarcoma, osteosarcoma, synovial sarcoma, rhabdomyosarcoma, undifferentiated sarcoma, reticulum cell sarcoma, neurofibrosarcoma, and malignant fibrous histiocytoma. Cardiac tumors produce a large variety of symptoms through any of 4 mechanisms. Their mass can obstruct intracardiac blood flow or interfere with valve function. Local invasion can lead to arrhythmias or pericardial effusions with tamponade. Bits of tumor can embolize, causing systemic deficits when the tumors are on the left side of the heart. Finally, the tumors may cause systemic or constitutional symptoms. Some tumors, of course, produce no symptoms and become evident as incidental findings. The most useful diagnostic tool is the echocardiogram, which in almost all cases precisely locates the tumor and defines its extent. The echocardiographic appearance may also allow quite accurate prediction of the tumor type and whether it is malignant or benign. Magnetic resonance imaging serves as the next most important test where the density of T1 and T2 images may allow tumor cell type identification. With few exceptions, these tumors require operative excision. Most benign tumors can be resected completely; a few, because of their large size, cannot be, and only tumor debulking may be possible. Heart transplantation should be considered for these patients. Many of the malignant tumors cannot be resected completely, either because of the extent of local spread and invasion or because of the frequent distant metastases. Transplantation may also be an option for those with extensive local disease. The long-term results for resected benign tumors are excellent; the long-term results for sarcomas are very poor, and there are few survivors. For patients with unresectable sarcomas, radiation and chemotherapy may be used, but without great expectation of successful results. PMID- 10807432 TI - Papillary fibroelastomas. AB - Papillary fibroelastomas are rare benign neoplasms, predominantly involving cardiac valves, that have been discovered with increasing frequency through the use of echocardiography. Most are papillary lesions, less than 1 cm in size, connected to the valve or mural endocardium by a small stalk. Although often asymptomatic, embolization from the lesion or attached thrombus may cause serious neurological or cardiac events. All symptomatic papillary fibroelastomas should be removed unless there are compelling contraindications, in which case anticoagulation is an acceptable but unreliable alternative. Surgical removal is safe, simple, effective, and permanent. Asymptomatic lesions of the left side of the heart should be removed because of their potentially serious or fatal consequences, whereas those arising from the right side of the heart may be observed. PMID- 10807433 TI - Surgery for tumors with cavoatrial extension. AB - Intracardiac extension of infradiaphragmatic tumors is an uncommon but significant surgical challenge for the treating surgeon. Renal cell carcinoma is the most common malignant tumor seen, with Wilms' tumor, uterine tumors (both benign and malignant), adrenal tumors, hepatoma, and lymphoma less frequently encountered. Surgical resection requires involvement of a cardiothoracic surgeon, urologist, and/or gynecologist. Cardiopulmonary bypass and deep hypothermic circulatory arrest provide the safest and most effective technique for removing these tumors. PMID- 10807434 TI - Malignancies involving the pericardium. AB - Malignancies rarely arise from the pericardium. Mesothelioma, the most common of these, is usually unresectable and almost always incurable. Malignancies may secondarily involve the pericardium by direct extension. Carcinoma of the lung and malignant thymoma with limited direct invasion of the pericardium both can undergo complete and potentially curative resections, but adjuvant therapy is usually indicated. More frequently, malignancies involve the pericardium by a process of retrograde lymphangitic spread or hematogenous dissemination. These patients present with a symptomatic pericardial effusion and occasionally pericardial tamponade. Subxiphoid pericardiostomy and drainage is a safe procedure that provides effective and durable symptomatic relief in these terminally ill patients. More aggressive open procedures should be reserved for loculated or recurrent pericardial effusions. PMID- 10807435 TI - An overview of HIV infection and AIDS: etiology, pathogenesis, diagnosis, epidemiology, and occupational exposure. AB - Thanks to a worldwide collaborative effort among health care providers, academia, governments, and industry, our knowledge base about infection caused by the human immunodeficiency virus (HIV) has expanded exponentially. During the past 2 decades, we have learned about its pathogenesis, virology, immunology, epidemiology and treatment. In the developed world, the approach to persons with HIV disease has evolved from palliative disease care to use of a chronic disease model, where survival is measured by decades, not months or years. More and more, clinical decision-making for HIV-infected patients is driven by comorbidities, including cardiothoracic disease. Thus, our clinically stable HIV population is increasingly accessing those health care services required by any maturing population, including the usual services of cardiothoracic surgeons. In this article, we review the basic facts of HIV disease, with an emphasis on occupational risks and infection control procedures. PMID- 10807436 TI - Treatment of HIV disease in the new millennium. AB - Highly active antiretroviral therapy (HAART), which typically consists of 3-drug combinations of antiretroviral agents, has decreased dramatically the incidence of AIDS and death among HIV-infected persons in the United States. HIV infection no longer is viewed as a death sentence. Complete suppression of viral replication through the use of HAART can reverse the immune deficits formerly thought to be inevitable. However, drug toxicity, cross-resistance, and less-than perfect adherence to prescribed antiretroviral regimens make the medical management of the HIV-infected person complex. An overview of the current approach to antiretroviral therapy is presented in this article. PMID- 10807437 TI - Cardiac surgery and the human immunodeficiency virus. AB - Since the first deliberate open heart operation was performed on a patient known to be carrying HIV, much has been learned. The fear that cardiopulmonary bypass might cause acceleration of the disease has not been borne out. Patients infected with HIV have shown considerable tolerance to major cardiac and pulmonary surgery. Indeed, the extraordinary fruits of a massive research effort have made it reasonable to perform elective surgery and to offer major surgery to patients with the full-blown syndrome of AIDS. The concern that the operators would be exposed to significant risk of acquiring the infection during surgery has proved to be unfounded. This has been in part due to the widespread adoption of universal precautions against the passage of microorganisms from patient to operator. However, there remain surgeons who ignore these precautions. The price they pay is the risk of acquiring hepatitis, which is far more easily transmitted than AIDS and may be fatal. PMID- 10807438 TI - Thoracic malignancies associated with AIDS. AB - The immunocompromised state is a major risk factor for the development of malignant tumors. Individuals with human immunodeficiency virus (HIV), and acquired immunodeficiency syndrome (AIDS) represent a large segment of the immunocompromised group of patients. Kaposi's sarcoma, B-cell non-Hodgkin's lymphoma, primary central nervous system lymphoma, and invasive cervical carcinoma are malignant tumors that are all AIDS-defining illnesses. Lung cancer is also seen with a higher frequency in AIDS patients. Malignant tumors are more aggressive in this group of patients as compared with the general population. Prognosis is poor, although with the improved survivals seen with new treatment in these patients, aggressive therapy is still warranted. PMID- 10807439 TI - Phytoestrogens and cardiovascular health. AB - Coronary artery disease is the leading overall cause of mortality for women and increases dramatically after menopause. Estrogen has many beneficial cardiovascular actions although concerns have been raised about its effects on the progression of breast and uterine neoplasms and its tendency to increase coagulability. Selective estrogen agonists may be superior to conventional estrogens. A dietary source of a partial estrogen agonist is the plant-based group of phytoestrogens, which include isoflavones, lignans and coumestans. Phytoestrogens have a similar structure to estradiol and have weak affinity for the estrogen receptor. Epidemiologic data indicate that women ingesting high amounts of phytoestrogens, particularly as isoflavones in soy products, have less cardiovascular disease, breast and uterine cancer and menopausal symptoms than those eating Western diets. Preclinical and clinical studies have found that isoflavones have lipid-lowering effects as well as the ability to inhibit low density lipoprotein oxidation. They have been shown to normalize vascular reactivity in estrogen-deprived primates. Furthermore, phytoestrogens have antineoplastic effects with inhibition of cellular proliferation as well as angiogenesis, properties that could be protective against cancer development. Finally, menopausal symptoms and bone density may be favorably influenced by phytoestrogens. In summary, phytoestrogens, in the form of dietary isoflavones, represent a new area to explore in pursuit of nutritional approaches to cardiovascular protection. PMID- 10807440 TI - Effective prevention of atrial fibrillation by continuous atrial overdrive pacing after coronary artery bypass surgery. AB - OBJECTIVES: The present study was aimed to evaluate the efficacy of a specific algorithm with continuous atrial dynamic overdrive pacing to prevent atrial fibrillation (AF) after coronary artery bypass graft (CABG) surgery. BACKGROUND: Atrial fibrillation occurs in 30% to 40% of patients after cardiac surgery with a peak incidence on the second day. It still represents a challenge for postoperative prevention and treatment and may have medical and cost implications. METHODS: Ninety-six consecutive patients undergoing CABG for severe coronary artery disease and in sinus rhythm without antiarrhythmic therapy on the second postoperative day were randomized to have or not 24 h of atrial pacing through temporary epicardial wires using a permanent dynamic overdrive algorithm. Holter ECGs recorded the same day in both groups were analyzed to detect AF occurrence. RESULTS: No difference was observed in baseline data between the two study groups, particularly for age, male gender, history of AF, ventricular function, severity of coronary artery disease, preoperative beta-adrenergic blocking agent therapy or P-wave duration. The incidence of AF was significantly lower (p = 0.036) in the paced group (10%) compared with control subjects (27%). Multivariate analysis showed AF incidence to increase with age (p = 0.051) but not in patients with pacing (p = 0.078). It decreased with a better left ventricular ejection fraction only in conjunction with atrial pacing (p = 0.018). CONCLUSIONS: We conclude that continuous atrial pacing with an algorithm for dynamic overdrive reduces significantly incidence of AF the second day after CABG surgery, particularly in patients with preserved left ventricular function. PMID- 10807441 TI - Atrial pacing for the prevention of atrial fibrillation after cardiovascular surgery. AB - OBJECTIVE: The purpose of this study was to determine the efficacy of atrial pacing in the prevention of atrial fibrillation following cardiovascular surgery. BACKGROUND: Although pharmacologic therapy has been used to help prevent postoperative atrial fibrillation, it suffers from limited efficacy and adverse effects. In the nonoperative setting, novel pacing strategies have been shown to reduce recurrences of atrial fibrillation and prolong arrhythmia-free periods in patients with paroxysmal atrial arrhythmias. METHODS: A total of 154 patients (115 men; mean age, 65 +/- 10 years; ejection fraction, 53 +/- 10%) undergoing cardiac surgery (coronary artery bypass surgery, 88.3%; aortic valve replacement, 4.5%; coronary bypass + aortic valve replacement, 7.1%) had right and left atrial epicardial pacing electrodes placed at the time of surgery. Patients were randomized to either no pacing, right atrial (RAP), left atrial (LAP) or biatrial pacing (BAP) for 72 h after surgery. Beta-adrenergic blocking agents were administered concurrently to all patients following surgery. RESULTS: There was a reduction in the incidence of postoperative atrial fibrillation from 37.5% in patients receiving no postoperative pacing to 17% (p < 0.005) in patients assigned to one of the three pacing strategies. The length of hospital stay was reduced by 22% from 7.8 +/- 3.7 days to 6.1 +/- 2.3 days (p = 0.003) in patients assigned to postoperative atrial pacing. The incidence of atrial fibrillation was lower in each of the paced groups (RAP, 8%; LAP, 20%; BAP, 26%) compared with patients who did not receive postoperative pacing (37.5%). CONCLUSION: Postoperative atrial pacing, in conjunction with beta-blockade, significantly reduced both the incidence of atrial fibrillation and the length of hospital stay following cardiovascular surgery. Additional studies are needed to determine the most effective anatomic pacing site. PMID- 10807442 TI - Atrial pacing for the prevention of postoperative atrial fibrillation: how and where to pace? PMID- 10807443 TI - Spontaneous episodes of atrial fibrillation after implantation of the Metrix Atrioverter: observations on treated and nontreated episodes. Metrix Investigators. AB - OBJECTIVES: We sought to evaluate the number and duration of device-treated and self-terminating, nontreated episodes of atrial fibrillation (AF) after implantation of the Metrix Atrioverter. BACKGROUND: A recent study has shown that the Atrioverter can rapidly restore sinus rhythm in patients with AF; however, the effect of the device on the clinical course of the arrhythmia in these patients is unknown. METHODS: The Atrioverter was implanted in 51 patients with symptomatic, recurrent, drug-refractory AF. The device was programmed to periodically monitor the cardiac rhythm. Defibrillation of AF episodes was performed under physician observation. RESULTS: During a mean follow-up of 260 +/ 144 days, 1,161 episodes of AF were observed during valid monitoring periods in 45 of 51 patients. Forty-one patients experienced 231 episodes for which they sought defibrillation therapy. The average duration of the treated episodes during valid monitoring periods (190 of 231 episodes in 39 of 41 patients) was significantly longer than that of the nontreated episodes (38 +/- 44 vs. 10 +/- 8 h; p < 0.05). The time between episodes requiring Atrioverter therapy increased, and the risk of having an episode requiring treatment decreased. No changes were observed in the number and duration of the short-lasting, nontreated episodes as time since implantation of the device increased. CONCLUSIONS: In patients with symptomatic, recurrent, drug-refractory AF, the frequency of long-lasting episodes, which were treated under observation with repeated defibrillation using the Atrioverter, decreased. The number and duration of short-lasting, nontreated episodes did not change during the 20-month study period. The effect of ambulatory use of the device on the recurrence of short-lasting episodes needs to be evaluated. PMID- 10807444 TI - Intravenous sotalol decreases transthoracic cardioversion energy requirement for chronic atrial fibrillation in humans: assessment of the electrophysiological effects by biatrial basket electrodes. AB - OBJECTIVES: This study was undertaken to assess the effects of sotalol on the transthoracic cardioversion energy requirement for chronic atrial fibrillation (AF) and on the atrial electrograms during AF recorded by two basket electrodes. BACKGROUND: The effects of sotalol infusion on transthoracic electrical cardioversion for chronic atrial fibrillation in humans have not been well investigated. METHODS: We included 18 patients with persistent AF for more than three months. Atrial electrograms were recorded by two basket electrodes positioned in each atrium respectively. Transthoracic cardioversion was performed before and after sotalol 1.5 mg/kg i.v. infusion. RESULTS: In the 14 patients whose AF could be terminated by cardioversion before sotalol infusion, the atrial defibrillation energy was significantly reduced after sotalol infusion (236 +/- 74 jules [J] vs. 186 +/- 77 J; p < 0.01). Atrial fibrillation was refractory to cardioversion in four patients at baseline and was converted to sinus rhythm by cardioversion after sotalol infusion in two of them. We further divided the patients into two groups. Group A consisted of 10 patients in whom the energy requirement was decreased by sotalol while group B consisted of eight patients in whom the energy requirement was not decreased. The mean A-A (atrial local electrogram) intervals during AF were significantly increased after sotalol infusion in both groups, but the increment of A-A interval was significantly larger in group A than it was in group B patients (36 +/- 13 ms vs. 22 +/- 8 ms for the right atrium; 19 +/- 7 ms vs. 9 +/- 7 ms for the left atrium; both p < 0.05). The spatial and temporal dispersions of A-A intervals were not significantly changed after sotalol infusion in both atria in both groups. CONCLUSIONS: Sotalol decreases the atrial defibrillation energy requirement by increasing atrial refractoriness but not by decreasing the dispersion of refractoriness. PMID- 10807445 TI - Nonsurgical transthoracic epicardial catheter ablation to treat recurrent ventricular tachycardia occurring late after myocardial infarction. AB - OBJECTIVES: We sought to evaluate feasibility, safety and results of transthoracic epicardial catheter ablation in patients with ventricular tachycardia occurring late after an inferior wall myocardial infarction. BACKGROUND: Transthoracic epicardial catheter ablation effectively controls recurrent ventricular tachycardia (VT) in patients with Chagas' disease in whom epicardial circuits predominate. Epicardial circuits also occur in postinfarction VT. METHODS: Fourteen consecutive patients aged 53.6 +/- 14.5 years with postinfarction VT related to the inferior wall were studied. The VT cycle length was 412 +/- 51 ms. Two patients had previously undergone unsuccessful standard endocardial radiofrequency energy (RF) ablation. The VT was incessant in one patient. Left ventricular angiography showed inferior akinesia in 13 patients and an inferior aneurysm in 1 patient. Ablation was performed with a regular steerable catheter placed into the pericardial sac by pericardial puncture. RESULTS: The pericardial space was reached in all patients. Electrophysiologic evidence of an epicardial circuit was present in 7 of 30 VTs. Due to a high stimulation threshold, empirical thermal mapping was the only criterion used to select the site for ablation. Three VTs were interrupted during the first RF pulse. Two pulses were necessary to render it noninducible in 3 patients (1 VT per patient). In the remaining 4 VTs, 3, 3, 4 and 5 RF pulses, respectively, were used. The overall success was 37.14% (95% confidence interval, 11.83% to 62.45%). Patients are asymptomatic for 14 +/- 2 months. CONCLUSIONS: Postinfarction pericardial adherence does not preclude epicardial mapping and ablation to control VT related to an epicardial circuit in postinferior wall myocardial infarction. PMID- 10807446 TI - Epicardial approach to the ablation of ventricular tachycardia in coronary artery disease: an alternative or ancillary approach. PMID- 10807447 TI - Regional myocardial blood flow in patients with sick sinus syndrome randomized to long-term single chamber atrial or dual chamber pacing--effect of pacing mode and rate. AB - OBJECTIVES: This study aimed to evaluate regional myocardial blood flow (MBF) and global left ventricular ejection fraction (LVEF) during chronic pacing in patients with sick sinus syndrome (SSS) randomized to either single chamber atrial (AAI) or dual chamber (DDD) pacing. BACKGROUND: Experimental studies indicate that chronic pacing in the right ventricular apex changes regional MBF, thereby compromising left ventricular function. METHODS: Thirty patients (age 74 +/- 10 years) were randomized to AAI (n = 15) or DDD (n = 15) pacemakers. After 22 +/- 7 months of pacing, MBF was quantified with 13N-labeled ammonia positron emission tomography scanning at 60 beats per min and 90 beats per min. Patients in the DDD group furthermore underwent MBF measurement at temporary AAI pacing, 60 beats per min. Myocardial blood flow was assessed in the anterior, lateral, inferior and septal regions, and the global mean MBF was calculated. Left ventricular ejection fraction was determined by echocardiography at pacemaker implantation and at the time of MBF measurements. RESULTS: Myocardial blood flow at rates 60 and 90 beats per min did not differ between the AAI and DDD groups. During temporary AAI pacing in the DDD group, MBF was significantly higher than during DDD pacing in both the inferior (p = 0.001) and septal (p = 0.004) regions and also globally (0.61 +/- 0.15 vs. 0.53 +/- 0.13 mL x g(-1) x min(-1), p = 0.005). In the DDD group, LVEF decreased from pacemaker implantation to time of MBF measurements (0.61 +/- 0.09 vs. 0.56 +/- 0.07, p = 0.013). Left ventricular ejection fraction during temporary AAI pacing at time of MBF measurements was not different from LVEF at pacemaker implantation. CONCLUSIONS: In patients with SSS, chronic DDD pacing reduced inferior, septal and global mean MBF as well as LVEF, as compared with temporary AAI pacing. The LVEF reversed to baseline level during temporary AAI pacing despite 22 months of permanent ventricular pacing preceding it. Augmenting pace rate to 90 beats per min increased MBF equally in the two treatment groups. PMID- 10807448 TI - Effects of controlled breathing, mental activity and mental stress with or without verbalization on heart rate variability. AB - OBJECTIVES: To assess whether talking or reading (silently or aloud) could affect heart rate variability (HRV) and to what extent these changes require a simultaneous recording of respiratory activity to be correctly interpreted. BACKGROUND: Sympathetic predominance in the power spectrum obtained from short- and long-term HRV recordings predicts a poor prognosis in a number of cardiac diseases. Heart rate variability is often recorded without measuring respiration; slow breaths might artefactually increase low frequency power in RR interval (RR) and falsely mimic sympathetic activation. METHODS: In 12 healthy volunteers we evaluated the effect of free talking and reading, silently and aloud, on respiration, RR and blood pressure (BP). We also compared spontaneous breathing to controlled breathing and mental arithmetic, silent or aloud. The power in the so called low- (LF) and high-frequency (HF) bands in RR and BP was obtained from autoregressive power spectrum analysis. RESULTS: Compared with spontaneous breathing, reading silently increased the speed of breathing (p < 0.05), decreased mean RR and RR variability and increased BP. Reading aloud, free talking and mental arithmetic aloud shifted the respiratory frequency into the LF band, thus increasing LF% and decreasing HF% to a similar degree in both RR and respiration, with decrease in mean RR but with minor differences in crude RR variability. CONCLUSIONS: Simple mental and verbal activities markedly affect HRV through changes in respiratory frequency. This possibility should be taken into account when analyzing HRV without simultaneous acquisition and analysis of respiration. PMID- 10807449 TI - Distinct hemodynamic profiles in patients with vasovagal syncope: a heterogeneous population. AB - OBJECTIVE: The objective was to investigate mechanisms of vasovagal syncope by identifying laboratory techniques that characterize cardiovascular profiles in patients with vasovagal syncope. BACKGROUND: The triggering mechanisms of vasovagal syncope are complex. The patient population is likely heterogeneous. We hypothesized that distinct hemodynamic profiles are definable with provocative maneuvers. METHODS: Three groups of subjects were matched for age and gender: 16 patients with a history of syncope and an inducible vasovagal response during passive tilt table testing (70 degrees, 45 min, group I), 16 with a history of syncope, negative passive tilt table testing but positive isoproterenol tilt table testing (0.05 microg/kg per min, 70 degrees, 10 min, group II), and 16 control subjects. Beat-to-beat hemodynamic functions were determined noninvasively by photo-plethysmography and impedance cardiography. RESULTS: At baseline, hemodynamic functions were not different among the three groups (supine). In response to tilt before any symptoms developed, total peripheral resistance decreased 9% +/- 14% in group I from baseline supine to tilt position but increased 27% +/- 18% in group II and 28% +/- 17% in controls (p < 0.001). Responses to isoproterenol were not significantly different between group II and controls in supine position. In response to tilt during isoproterenol infusion before any symptoms developed, total peripheral resistance decreased 24% +/- 20% in group II and increased 20% +/- 48% in controls (p = 0.002). CONCLUSIONS: Group I patients may have impaired ability to increase vascular resistance during orthostatic stress. The inability to overcome isoproterenol-induced vasodilatation during tilt is important in triggering a vasovagal response in group II patients. These data suggest that the population with vasovagal response is heterogeneous. Distinct hemodynamic profiles in response to various provocative maneuvers are definable with noninvasive, continuous monitoring techniques. PMID- 10807450 TI - Tracking dynamic conduction recovery across the cavotricuspid isthmus. AB - OBJECTIVES: We sought to assess the dynamic temporal course of conduction recovery during and after radiofrequency (RF) catheter ablation of the cavotricuspid isthmus. BACKGROUND: Although cavotricuspid isthmus block is accepted as the best end point of ablation for typical flutter, conduction recovery is thought to underlie many eventual recurrences. Its time course and frequency have not been determined. METHODS: In a prospective group of 30 patients (26 men and 4 women, age 64 +/- 12 years) undergoing ablation of typical flutter in the cavotricuspid isthmus, the morphology of the P wave during pacing from the low lateral right atrium after achievement of complete isthmus block was identified as a reference. Regression of this morphologic P wave change was confirmed to be associated with intracardiac evidence of the recovery of cavotricuspid isthmus conduction and was observed throughout the procedure both during ablation in sinus rhythm (n = 15, group B) and just after flutter termination (n = 15, group A). RESULTS: Stable complete isthmus block was achieved in all patients; 29 had a terminal positivity of the paced P wave. Flutter termination resulted in stable block and terminal P wave positivity in three patients, transient terminal P wave positivity and transient block despite continuing RF at the same site in five patients and no block in the remaining seven patients. Conduction recovery identified by recovery of P wave changes was nearly as common (48%) during ablation in sinus rhythm. Multiple recoveries were noted in some patients, and 72% of all recoveries occurred within 1 min. Conduction recovery was only rarely associated with coagulum, impedance elevation or pops. CONCLUSIONS: Conduction recovery in the cavotricuspid isthmus is common during and after ablation and can be accurately, dynamically and continuously observed by monitoring the recovery of the low lateral right atrial paced P wave change. PMID- 10807451 TI - Exploiting rate-related hysteresis in repolarization alternans to improve risk stratification for ventricular tachycardia. AB - OBJECTIVES: We sought to study the effect of heart rate acceleration and deceleration on the ability of repolarization alternans (RPA) to stratify ventricular tachycardia (VT) risk. BACKGROUND: Heart rate fluctuations alter arrhythmic propensity, yet it is unclear whether fluctuations, as well as absolute rate, dynamically increase VT risk. We hypothesized that repolarization heterogeneity reflected by RPA would exhibit hysteresis during rising and falling heart rate, which may reflect arrhythmic propensity. METHODS: The RPA magnitude (absolute voltage of alternation [V(alt)] and T-wave alternans ratio [TWAR]) and temporal distribution were determined from the electrocardiogram (ECG) in 60 patients during paced heart rate acceleration from 100 to 150 beats/min, then deceleration to 100 beats/min at electrophysiologic study (EPS). The V(alt) and TWAR thresholds were varied prospectively to generate receiver-operating characteristics (ROC) for the prediction of inducible VT at EPS. RESULTS: Thirty six patients were induced into VT and 24 were not. Hysteresis of RPA was seen. The V(alt) reached steady-state within 60 beats of each rate transition and was higher in deceleration than in acceleration at matched heart rates. In induced patients, V(alt) rose then fell with heart rate. In noninduced patients, V(alt) was insensitive to acceleration, but rose on initial deceleration. The RPA distributed later within repolarization in induced patients but, on deceleration, moved earlier in both groups. By ROC analysis, V(alt) = 2.6 microV in late repolarization at 120 beats/min provided optimal sensitivity and specificity for VT in acceleration (87.5% and 88.7%, respectively) versus deceleration (80% and 62.5%, respectively; p = 0.004, chi-square test). CONCLUSIONS: 1) Physiologic fluctuations in heart rate may affect the clinical utility of RPA for VT risk stratification; and 2) repolarization dispersion measured by RPA is more exaggerated during deceleration than acceleration at matched heart rates (rate hysteresis). PMID- 10807452 TI - Clinical profile of congenital coronary artery anomalies with origin from the wrong aortic sinus leading to sudden death in young competitive athletes. AB - OBJECTIVES: The purpose of this study is to characterize the clinical profile and identify clinical markers that would enable the detection during life of anomalous coronary artery origin from the wrong aortic sinus (with course between the aorta and pulmonary trunk) in young competitive athletes. BACKGROUND: Congenital coronary artery anomalies are not uncommonly associated with sudden death in young athletes, the catastrophic event probably provoked by myocardial ischemia. Such coronary anomalies are rarely identified during life, often because of insufficient clinical suspicion. However, since anomalous coronary artery origin is amenable to surgical treatment, timely clinical identification is crucial. METHODS: Because of the paucity of available data characterizing the clinical profile of wrong sinus coronary artery malformations, we reviewed two large registries comprised of young competitive athletes who died suddenly, assembled consecutively in the U.S. and Italy. RESULTS: We reported 27 sudden deaths in young athletes, identified solely at autopsy and due to either left main coronary artery from the right aortic sinus (n = 23) or right coronary artery from the left sinus (n = 4). Each athlete died either during (n = 25) or immediately after (n = 2) intense exertion on the athletic field. Fifteen athletes (55%) had no clinical cardiovascular manifestations or testing during life. However, in the remaining 12 athletes (45%) aged 16 +/- 7, certain clinical data were available. Premonitory symptoms had occurred in 10, including syncope in four (exertional in three and recurrent in two, 3 to 24 months before death) and chest pain in five (exertional in three, all single episodes, < or =24 months before death). All cardiovascular tests were within normal limits, including 12 lead electrocardiogram (ECG) pattern (in 9/9), stress ECG with maximal exercise (in 6/6) and left ventricular wall motion and cardiac dimensions by two dimensional echocardiography (in 2/2). CONCLUSIONS: With regard to congenital coronary artery anomalies of wrong aortic sinus origin in young competitive athletes, 1) standard testing with ECG under resting or exercise conditions is unlikely to provide clinical evidence of myocardial ischemia and would not be reliable as screening tests in large athletic populations, 2) premonitory cardiac symptoms not uncommonly occurred shortly before sudden death (typically associated with anomalous left main coronary artery), suggesting that a history of exertional syncope or chest pain requires exclusion of this anomaly. These observations have important implications for the preparticipation screening of competitive athletes. PMID- 10807453 TI - Diabetes mellitus and outcome after primary coronary angioplasty for acute myocardial infarction: lessons from the GUSTO-IIb Angioplasty Substudy. Global Use of Strategies to Open Occluded Arteries in Acute Coronary Syndromes. AB - OBJECTIVES: We sought to compare the efficacy of primary angioplasty in diabetics versus nondiabetics and to evaluate the relative benefits of angioplasty over thrombolytic therapy among diabetics. BACKGROUND: Primary angioplasty for myocardial infarction is at least as effective as thrombolytic therapy in the general population. However, the influence of diabetic status on outcome after primary angioplasty versus thrombolysis remains unknown. METHODS: Patients in the Global Use of Strategies To Open Occluded Arteries in Acute Coronary Syndromes (GUSTO-IIb) Angioplasty Substudy were randomized to receive either primary angioplasty or accelerated alteplase. The interaction of diabetic status (diabetics n = 177, nondiabetics n = 961) and treatment strategy with the occurrence of the primary end point (death, nonfatal reinfarction or nonfatal, disabling stroke at 30 days) was analyzed (power to detect a 40% relative reduction in the primary end point with alpha = 0.05 and beta = 0.20). Among patients who were randomized to and underwent primary angioplasty, procedural success (defined as residual stenosis <50% and TIMI grade 3 flow) was assessed based on diabetic status. RESULTS: Compared with nondiabetics, diabetics had worse baseline clinical and angiographic profiles. Despite more severe stenosis and poorer flow in the culprit artery, procedural success with angioplasty was similar for diabetics (n = 81; 70.4%) and nondiabetics (n = 391; 72.4%). Outcome at 30 days was better for nondiabetics randomized to angioplasty versus alteplase (adjusted odds ratio, 0.62; 95% confidence interval, 0.41-0.96) with a similar trend for diabetics (0.70, [0.29-1.72]). We noted no interaction between diabetic status and treatment strategy on outcome (p = 0.88). CONCLUSIONS: Primary angioplasty was similarly successful in diabetics and nondiabetics and appeared to be more effective than thrombolytic therapy among diabetics with acute infarction. PMID- 10807454 TI - Acute myocardial infarction: are diabetics different? PMID- 10807455 TI - Relationship between corrected TIMI frame counts at three weeks and late survival after myocardial infarction. AB - OBJECTIVES: To evaluate the corrected Thrombolysis in Myocardial Infarction (TIMI) frame count (CTFC) as a predictor of late survival after myocardial infarction. BACKGROUND: Thrombolysis in Myocardial Infarction flow grades predict late survival after myocardial infarction. The CTFC provides a more reproducible measurement of infarct-related artery blood flow than the TIMI flow grade, and has been linked to 30-day outcomes, but it has not yet been established how the CTFC correlates with late survival. METHODS: Of 1,001 patients with acute myocardial infarction presenting within 4 h of symptom onset, 882 underwent angiography at approximately three weeks. Infarct artery flow was assessed, blinded to clinical outcomes, according to the CTFC and TIMI flow grade. Late cardiac mortality and survival were determined in 97.5% of patients. RESULTS: The mean CTFC was 40 +/- 29 in 644 patent infarct arteries (median, 34 [interquartile range, 24 to 47]). The CTFC, assessed as a continuous univariate variable, was found to be a predictor of five-year survival, as was the TIMI flow grade (both p < 0.001). On multivariate analysis, factors associated with five-year survival included the ejection fraction or end-systolic volume index (both p < 0.001); exercise duration (p = 0.005), age (p = 0.008), diabetes (p = 0.02) and CTFC (p = 0.02) or TIMI flow (p = 0.02). The same factors, except for the CTFC and TIMI flow grade, were predictors of 10-year survival. CONCLUSIONS: The CTFC three weeks after myocardial infarction was an independent predictor of five-year survival, but not 10-year survival. Although the CTFC provided additional prognostic information within TIMI flow grades, its superiority was not demonstrated. PMID- 10807456 TI - Remote myocardial dysfunction after acute anterior myocardial infarction: impact of left ventricular shape on regional function: a magnetic resonance myocardial tagging study. AB - OBJECTIVES: We sought to evaluate regional morphology and function in patients in their first week after having a reperfused anterior myocardial infarction (MI) using magnetic resonance (MR) myocardial tagging. BACKGROUND: The mechanism of myocardial dysfunction in the remote, noninfarct-related regions is an unresolved issue to date. METHODS: Sixteen patients with a first reperfused transmural anterior MI were studied with MR tagging at 5 +/- 2 days after the event, and the results were compared with those of an age-matched control group regions. The left ventricle (LV) was divided into infarct, adjacent and remote regions. Magnetic resonance tagging provided information on the regional ventricular morphology and function. RESULTS: Morphologically, an increase of the circumferential radius of curvature was found in the remote myocardium, whereas the longitudinal radius of curvature was increased in all regions of the LV. A significant increase in apical sphericity was also found. A significant reduction in strain and function was found not only in the infarct region, but also in the adjacent and remote myocardium. The loss in regional ejection fraction in the remote myocardium (61.4 +/- 11.7% in patients vs. 68.7 +/- 10.0% in control subjects, p < 0.0001) was related to a significant reduction of the longitudinal and circumferential strain, whereas systolic wall thickening was preserved. CONCLUSIONS: Remote myocardial dysfunction contributes significantly to the loss in global ventricular function. This could be secondary to morphologic changes in the infarct region, leading to an increased systolic longitudinal wall stress without loss of intrinsic contractility in the remote regions. PMID- 10807457 TI - Predictive value of C-reactive protein and troponin T in patients with unstable angina: a comparative analysis. CAPTURE Investigators. Chimeric c7E3 AntiPlatelet Therapy in Unstable angina REfractory to standard treatment trial. AB - OBJECTIVES: We evaluated C-reactive protein (CRP) and troponin T (TnT) for predicting six-month cardiac risk in patients with unstable angina. BACKGROUND: Troponin T is predictive of cardiac risk in patients with unstable angina. The clinical implications of elevated CRP in such patients remains controversial. METHODS: Baseline TnT and CRP values were determined in 447 patients with unstable angina enrolled in the placebo group of the Chimeric c7E3 AntiPlatelet Therapy in Unstable angina REfractory to standard treatment trial (CAPTURE) trial. All patients underwent a coronary intervention and were followed for a six month period in which 13 deaths and 47 myocardial infarctions were documented (MIs). RESULTS: Troponin T was >0.1 microg/liter in 30% and CRP was >10 mg/L in 41% of the patients. For the initial 72-h period (including coronary intervention), TnT (17.4% vs. 4.2%; p < 0.001) but not CRP (10.3% vs. 8%; p = 0.41) was predictive of mortality and MI. The TnT-positive patients displayed more frequent recurrent instability before the planned intervention (44.8% vs. 16.9%; p < 0.001), but in the CRP-positive patients, no such increase was observed (25.9% vs. 24.8%; p = 0.92). In contrast, for the six month follow-up period, CRP was predictive of cardiac risk (mortality, MI) (18.9% vs. 9.5%; p = 0.003). Using multivariate analysis, both CRP and TnT emerged as independent predictors of mortality and MI at six-month follow-up. Furthermore, the incidence of coronary restenosis during six-month follow-up was not related to TnT status (3% vs. 4.5%; p = 0.49); however, it was significantly related to CRP status (7% vs. 2.3%; p = 0.03). CONCLUSIONS: Troponin T, but not CRP, was predictive of cardiac risk during the initial 72-h period, whereas CRP was an independent predictor of both cardiac risk and repeated coronary revascularization (coronary artery bypass graft surgery and percutaneous transluminal coronary angioplasty) during six month follow-up. PMID- 10807458 TI - Unprotected left main coronary artery stenting: immediate and medium-term outcomes of 140 elective procedures. AB - OBJECTIVES: We sought to evaluate immediate and late outcomes after stenting for left main coronary artery (LMCA) stenosis. BACKGROUND: Conventional percutaneous transluminal coronary angioplasty (PTCA), for which coronary artery bypass grafting (CABG) has been the gold standard therapy for years, has yielded poor results in unprotected LMCA lesions. The development of coronary stents, together with their dramatic patency improvement provided by new antiplatelet regimens and their validation against restenosis, warrants a reappraisal of angioplasty in LMCA stenosis. METHODS: From January 1993 to September 1998, 140 consecutive unselected patients with unprotected LMCA stenosis underwent elective stenting. Group I included 47 high-CABG-risk patients, and group II included 93 low-CABG risk patients. Ticlopidine without aspirin was routinely started at least 72 h before the procedure and continued for one month. Patients were reevaluated monthly. A follow-up angiography was requested after six months. RESULTS: The procedure success rate was 100%. One-month mortality was 9% (4/47) in group I and 0% in group II. A follow-up angiography was obtained in 82% of cases, and target lesion revascularization was required in 17.4%. One-year actuarial survival was 89% in the first 29 group I patients and 97.5% in the first 63 group II patients. CONCLUSIONS: Stenting of unprotected LMCA stenosis provided excellent immediate results, particularly in good CABG candidates. Medium-term results were good, with a restenosis rate of 23%, similar to that seen after stenting at other coronary sites. Stenting deserves to be considered a safe and effective alternative to CABG in institutions performing large numbers of PTCAs. PMID- 10807459 TI - Is it time to offer elective percutaneous treatment of the unprotected left main coronary artery? PMID- 10807460 TI - Influence of treatment modality on angiographic outcome after coronary stenting in diabetic patients: a controlled study. AB - OBJECTIVES: This retrospective study was designed to determine the six-month angiographic outcome after stenting of native coronary arteries in insulin treated (ITDM) and non-ITDM patients with diabetes mellitus (DM) and compare the results with those in non-DM patients. BACKGROUND: The influence of the treatment modality for DM on restenosis in patients undergoing coronary artery stenting has not been elucidated sufficiently. METHODS: A total of 1,439 (70%) of 2,061 patients underwent repeated angiography within six months of coronary stenting. The ITDM and non-ITDM (oral hypoglycemic drugs or diet) were documented in 48 (3.3%) and 177 patients (12.3%), respectively, leaving 1,214 non-DM patients. RESULTS: Baseline reference vessel diameter tended to be smaller in ITDM patients (mean, 2.73 mm) than in non-DM and non-ITDM patients (2.88 mm and 2.85 mm, respectively). However, percent diameter stenosis was not different. The median number of stents deployed was 1; median stent length was 15 mm. Statistically significant differences were present after stenting for the means of minimal lumen diameter (MLD) and acute gain between ITDM patients (MLD: 2.67 mm, acute gain: 1.98 mm) and non-DM patients (MLD: 2.81 mm, acute gain: 2.16 mm). At follow up, percent diameter stenosis, late lumen loss and loss index were significantly higher in both non-ITDM lesions (42%, 1.14 mm and 0.56, respectively) and ITDM lesions (48%, 1.26 mm and 0.65, respectively) than in non-DM lesions (35%, 0.96 mm and 0.45, respectively). The corresponding differences between non-ITDM and ITDM lesions did not reach statistical significance. Restenosis rates in non-DM, non-ITDM and ITDM lesions were 23.8%, 32.8% (p = 0.013 vs. non-DM) and 39.6% (p = 0.02 vs. non-DM, p = 0.477 vs. non-ITDM), respectively. CONCLUSIONS: This study showed that compared with stenting in non-DM patients, stenting of native coronary arteries in DM patients is associated with significantly increased lumen renarrowing, regardless of the treatment modality for DM. PMID- 10807461 TI - Comparison of debulking followed by stenting versus stenting alone for saphenous vein graft aortoostial lesions: immediate and one-year clinical outcomes. AB - OBJECTIVES: We compared in-hospital and one-year clinical outcomes in patients undergoing debulking followed by stent implantation versus stenting alone for saphenous vein graft (SVG) aortoostial lesions. BACKGROUND: Stent implantation in SVG aortoostial lesions may improve procedural and late clinical outcomes. However, the impact of debulking before stenting in this complex lesion subset is unknown. METHODS: We studied 320 consecutive patients (340 SVG aortoostial lesions) treated with Palmaz-Schatz stents. Debulking with excimer laser or atherectomy was performed in 133 patients (139 lesions) before stenting (group I), while 187 patients (201 lesions) underwent stent implantation without debulking (group II). Procedural success and late clinical outcomes were compared between the groups. RESULTS: Overall procedural success (97.6%) was similar between the groups. Procedural complications were also similar (2.2% for group I and 2.6% for group II). At one-year follow-up, target lesion revascularization (TLR) was 19.4% for group I and 18.2% for group II (p = 0.47). There was no difference in cumulative death or Q wave myocardial infarction between the groups. Overall cardiac event-free survival was similar (69% for group I and 68% for group II). By Cox regression analysis, the independent predictors of late cardiac events were final lumen cross-sectional area (CSA) by intravascular ultrasound (IVUS) (p = 0.001) and restenotic lesions (p = 0.01). Similarly, final IVUS lumen CSA (p = 0.0001) and restenotic lesions (p = 0.006) were found to predict TLR at one year. CONCLUSIONS: These results suggest that, in most patients with SVG aortoostial lesions, debulking before stent implantation may not be necessary. PMID- 10807462 TI - In-stent restenosis: long-term outcome and predictors of subsequent target lesion revascularization after repeat balloon angioplasty. AB - OBJECTIVES: We sought to evaluate the long-term clinical outcome of patients undergoing successful balloon angioplasty for in-stent restenosis, and to determine correlates of the need for subsequent target lesion revascularization (TLR). BACKGROUND: In-stent restenosis can be safely treated by repeat percutaneous intervention. Reported subsequent TLR rates have varied from 20% to 80% and seem related to the type of restenotic lesion. METHODS: The study population comprised 234 patients with follow-up data who were successfully treated with repeat balloon angioplasty for in-stent restenosis in 257 lesions between May 1995 and January 1998 at our institution. RESULTS: Clinical follow-up was available at 459 (286 to 693) days after the repeat procedure. Event-free survival was 78.5% and 74.6% at 12 and 24 months, respectively. Recurrent events occurred in 58 patients (24.8%), including 6 deaths (2.6%), 4 myocardial infarction (1.7%) and repeat target vessel revascularization in 50 patients (21.4%). Independent predictors of repeat TLR were time to in-stent restenosis <90 days (Hazard ratio 4.67, p < 0.001), minimal luminal diameter after repeat procedure (Hazard ratio 0.38, p = 0.034) and the angiographic pattern of in-stent restenosis (Hazard ratio 1.65, p = 0.036). CONCLUSIONS: Balloon angioplasty is an effective means of treating in-stent restenosis. The long-term results are acceptable particularly for focal restenotic lesions. Further restenosis is more common in patients with early initial recurrence, more proliferative lesions and a poorer angiographic result from repeat angioplasty. PMID- 10807463 TI - In-stent restenosis: should an old device treat a new problem? PMID- 10807464 TI - Longitudinal straightening effect of stents is an additional predictor for major adverse cardiac events. Austrian Wiktor Stent Study Group and European Paragon Stent Investigators. AB - OBJECTIVES: The aim of this study was to perform an investigation of the effects of the longitudinal straightening of coronary arteries by stents and the possible association with major adverse cardiac events (MACE) (primary end point) and angiographic restenosis (secondary end point). BACKGROUND: Stent deployment straightens a tortuous artery, and any consequent arterial longitudinal stretch may contribute to MACE and stent restenosis severity. METHODS: Clinical, qualitative and quantitative angiographic data on 404 patients with single stent implantation were subjected to multivariate nominal logistic regression analysis for the prediction of MACE. The predictive accuracy, sensitivity and specificity values and cut-off points of the continuous variables were determined via receiver operating characteristics curves. The longitudinal straightening effect of stents was characterized through the changes in vessel angle (defined by the tangents to the proximal and distal parts of the stenoses/stents). RESULTS: Follow-up angiography on 354 patients revealed 73 cases of stent restenosis (> or =50% diameter stenosis). Coronary bypass surgery was performed in 4 patients and repeated percutaneous transluminal coronary angioplasty in 56 patients; acute myocardial infarction (AMI) occurred in 2 patients, and 4 patients died during the follow-up. The overall incidence of MACE (death, AMI and revascularization) was 16.3% (66 patients). The best predictive accuracies and sensitivities/specificities of factors indicative of MACE were found for the minimal lumen diameter (MLD) at follow-up (predictive accuracy: 0.9305, sensitivity/specificity: 86.6%), the post-stent MLD (0.773, 77.2%), the percent diameter stenosis (%DS) at follow-up (0.9432, 87.1%), the prestent vessel angulation (0.6797, 68.2%) and the poststent changes in vessel angulation (0.6279, 62.2%). Multivariate nominal logistic regression analysis demonstrated that a poststent MLD < or =2.63 mm (p = 0.0017, odds ratio [OR] = 17.961, 95% confidence interval [CI] = 17.45-20.428), an MLD at follow-up < or =1.7 mm (p = 0.0059, OR = 11.880, 95% CI = 11.490-14.093), a %DS at follow-up > or =42.2% (p = 0.0000, OR = 49.553, 95% CI = 48.024-53.507), a prestent vessel angulation > or =33.5 degrees (p = 0.0477, OR = 5.404, 95% CI = 5.382-7.142) and poststent changes in vessel angulation > or =9.1 degrees (p = 0.0026, OR = 19.161, 95% CI = 18.562-21.750) were significant predictors for MACE. Multiple linear regression revealed that the poststent MLD (multivariate p = 0.0001), the MLD at follow-up (p = 0.0000), the prestent vessel angulation (p = 0.0431) and the changes in vessel angulation after stent implantation (p = 0.0316) were significant independent variables predicting angiographic stent restenosis severity. CONCLUSIONS: The longitudinal straightening effect of coronary artery stents contributes significantly to the occurrence of MACE and angiographic restenosis, and this finding may have an impact on future stent design. PMID- 10807465 TI - Hemodynamic effects of immunoadsorption and subsequent immunoglobulin substitution in dilated cardiomyopathy: three-month results from a randomized study. AB - OBJECTIVES: The objective of our study was to assess the hemodynamic effects of immunoadsorption (IA) and subsequent immunoglobulin G (IgG) substitution in comparison with the effects of conventional medical treatment in patients with dilated cardiomyopathy (DCM). BACKGROUND: Various circulating cardiac autoantibodies have been detected among patients suffering from DCM. These antibodies are extractable by IA. METHODS: Patients with DCM (n = 18, New York Heart Association III-IV, left ventricular ejection fraction <30%) and who were on stable medication participated in the study. Hemodynamic measurements were performed using a Swan-Ganz thermodilution catheter. The patients were randomly assigned either to the treatment group with IA and subsequent IgG substitution (IA/IgG group, n = 9) or to the control group without IA/IgG (n = 9). In the IA/IgG group, the patients were initially treated in one IA session daily on three consecutive days. After the final IA session, 0.5 g/kg of polyclonal IgG was substituted. At one-month intervals, IA was then repeated for three further courses with one IA session daily on two consecutive days, until the third month. RESULTS: After the first IA course and IgG substitution, cardiac index (CI) increased from 2.1 (+/-0.1) to 2.8 (+/-0.1) L/min/m2 (p < 0.01) and stroke volume index (SVI) increased from 27.8 (+/-2.3) to 36.2 (+/-2.5) ml/m2 (p < 0.01). Systemic vascular resistance (SVR) decreased from 1,428 (+/-74) to 997 (+/-55) dyne x s x cm(-5) (p < 0.01). The improvement in CI, SVI and SVR persisted after three months. In contrast, hemodynamics did not change throughout the three months in the control group. CONCLUSIONS: Immunoadsorption and subsequent IgG substitution improves cardiovascular function in DCM. PMID- 10807466 TI - Early changes in left ventricular function in chronic asymptomatic alcoholics: relation to the duration of heavy drinking. AB - OBJECTIVES: This study sought to assess preclinical cardiac abnormalities in chronic alcoholic patients and possible differences among alcoholics related to the duration of heavy drinking. BACKGROUND: Chronic excessive alcohol intake has been reported as a possible cause of dilated cardiomyopathy. However, before the appearance of severe cardiac dysfunction, subtle signs of cardiac abnormalities may be identified. METHODS: We studied 30 healthy subjects (age 44 +/- 8 years) and 89 asymptomatic alcoholics (age 45 +/- 8 years, p = NS) divided into three groups, with short (S, 5-9 years, n = 31), intermediate (I, 10-15 years, n = 31) and long (L, 16-28 years, n = 27) duration of alcoholism. Transmitral early (E) and late (A) Doppler flow velocities, E/A ratio, deceleration time of E (DT) and isovolumic relaxation time (IVRT) were obtained. Left ventricular (LV) wall thickness and volumes were also determined by echocardiography, and LV mass and ejection fraction (EF) were calculated. RESULTS: The alcoholics had prolonged IVRT (92 +/- 11 vs. 83 +/- 7 ms, p < 0.001), longer DT (180 +/- 20 vs. 170 +/- 10 ms, p < 0.01), smaller E/A (1.25 +/- 0.34 vs. 1.40 +/- 0.32, p < 0.05), larger LV volumes (73 +/- 8 vs. 65 +/- 7 ml/m2, p < 0.001 for end-diastolic volume index; 25 +/- 4 vs. 21 +/- 2 ml/m2, p < 0.001 for end-systolic volume index), higher LV mass index (92 +/- 14 vs. 78 +/- 8 g/m2, p < 0.001) and thicker posterior wall (9 +/- 1 vs. 8 +/- 1 mm, p < 0.001). Ejection fraction did not differ between the two groups (66 +/- 4 vs. 67 +/- 2%). Deceleration time of the early transmitral flow velocity was longer in groups L (187 +/- 18 ms) and I (185 +/- 16 ms) compared with group S (168 +/- 17 ms, p < 0.001 for L and I vs. S), whereas A was higher in group L compared with S (43 +/- 10 vs. 51 +/- 10 cm/s, p < 0.005). Multiple regression analysis identified duration of heavy drinking as the most important variable affecting DT and A. CONCLUSIONS: Left ventricular dilation with preserved EF and impaired LV relaxation characterized LV function in chronic asymptomatic alcoholic patients. It appeared that the progression of abnormalities in LV diastolic filling related to the duration of alcoholism. PMID- 10807467 TI - Effects of bradykinin on coronary blood flow and vasomotion in transplant patients. AB - OBJECTIVES: To evaluate the effects of exogenous bradykinin on coronary epicardial and microcirculatory tone in transplant patients (HTXs), and to compare them with the effects of acetylcholine. BACKGROUND: Coronary endothelial dysfunction has been reported to occur early after heart transplantation, most notably when acetylcholine was the endothelium-function marker used. The effects of bradykinin on coronary vasomotion are unknown in HTXs. METHODS: Sixteen HTXs were compared 3.6 +/- 1.7 months after transplantation to seven control subjects. Coronary flow velocity was measured using guide-wire Doppler. Diameters (D) of three segments of the left coronary artery and coronary blood flow (CBF) were assessed at baseline, after 3-min infusions of increasing bradykinin doses (50, 150 and 250 ng/min) then of increasing acetylcholine doses (estimated blood concentrations of 10(-8), 10(-7) and 10(-6) M). RESULTS: Bradykinin induced similar dose-dependent increases in D and CBF in both groups: D was 11 +/- 12%, 19 +/- 14% and 22 +/- 16% (all p < 0.0001), and CBF was 50 +/- 40%, 130 +/- 68% and 186 +/- 77% (all p < 0.0001). Acetylcholine induced significant epicardial vasodilation in control subjects and vasoconstriction in HTX, as well as a marked increase in CBF in both groups. Acute allograft rejection, present in 8 of the 16 HTXs, did not modify responses to bradykinin, but was associated with a smaller CBF increase in response to acetylcholine (p < 0.05). CONCLUSIONS: The coronary vasodilating effects of bradykinin are preserved early after heart transplantation, even in the presence of acute allograft rejection. Although there is an abnormal vasoconstricting response to acetylcholine reflecting endothelium dysfunction, the endothelium remains a functionally active organ in heart transplant recipients. PMID- 10807468 TI - Oral vitamin C and endothelial function in smokers: short-term improvement, but no sustained beneficial effect. AB - OBJECTIVES: To test the hypothesis that antioxidant therapy would improve endothelial function in smokers. BACKGROUND: Several studies have documented a beneficial effect of short-term oral or parenteral vitamin C on endothelial physiology in subjects with early arterial dysfunction. Possible long-term effects of vitamin C on endothelial function, however, are not known. METHODS: We studied the effects of short- and long-term oral vitamin C therapy on endothelial function in 20 healthy young adult smokers (age 36 +/- 6 years, 8 male subjects, 21 +/- 10 pack-years). Each subject was studied at baseline, 2 h after a single dose of 2 g vitamin C and 8 weeks after taking 1 g vitamin C daily, and after placebo, in a randomized double-blind crossover study. Blood samples were analyzed for plasma ascorbate levels and endothelial function was measured as flow-mediated dilation of the brachial artery, using high resolution ultrasound. Nitroglycerin-mediated dilation (endothelium-independent) was also measured at each visit. RESULTS: At baseline, plasma ascorbate level was low in the smokers (42 +/- 21 micromol/liter; normal range, 50 to 150 micromol/liter), increased with vitamin C therapy after 2 h to 120 +/- 54 micromol/liter (p < 0.001) and remained elevated after eight weeks of supplementation at 92 +/- 32 micromol/liter (p < 0.001, compared with placebo). Flow-mediated dilation, however, increased at 2 h (from 2.8 +/- 2.0% to 6.3 +/- 2.8%, p < 0.001), but there was no sustained beneficial effect after eight weeks (3.9 +/- 3.2%, p = 0.26). Nitroglycerin-mediated dilation was unchanged throughout. CONCLUSION: Oral vitamin C therapy improves endothelial dysfunction in the short term in healthy young smokers, but it has no beneficial long-term effect, despite sustained elevation of plasma ascorbate levels. PMID- 10807469 TI - Inhibitory effects of low-density lipoproteins from men with type II diabetes on endothelium-dependent relaxation. AB - OBJECTIVES: The object of the present study is to determine whether native (n) low-density lipoprotein (LDL) isolated from men with type II diabetes and abnormal endothelial function inhibits endothelium-dependent relaxation more than n-LDL isolated from nondiabetic control subjects. BACKGROUND: Endothelium dependent vasodilation is impaired in men with type II diabetes and this may result from qualitative rather than quantitative abnormalities of LDL. METHODS: Forearm blood flow responses to brachial artery infusions of acetylcholine (endothelium-dependent vasodilator) and nitroprusside (endothelium-independent vasodilator) were measured in 10 men with uncomplicated type II diabetes and 10 nondiabetic men of similar age and with similar plasma concentrations of LDL cholesterol. Native LDL was isolated by discontinuous density gradient ultracentrifugation using EDTA to prevent oxidation. Preconstricted rabbit aortic ring bioassay was used to determine inhibitory properties of n-LDL on endothelium dependent relaxation by measuring relaxation to acetylcholine (and nitroprusside) in the presence and absence of n-LDL. RESULTS: Forearm blood flow responses to acetylcholine but not nitroprusside were significantly impaired (p < 0.01) in diabetic men compared with control subjects. Native LDL (10 and 100 microg protein/ml) from diabetic men inhibited relaxation to acetylcholine by 13.9 +/- 4.8% and 61.9 +/- 7.8% (mean inhibition for all doses +/- SE), respectively, whereas n-LDL from control subjects inhibited relaxation by 7.3 +/- 3.0% and 23.9 +/- 5.7% (p < 0.01 for a difference between diabetic and control n-LDL). Relaxation to nitroprusside was not significantly inhibited by n-LDL. CONCLUSIONS: A qualitative abnormality of LDL may account for endothelial dysfunction in men with type II diabetes. PMID- 10807470 TI - Predictors of congestive heart failure in the elderly: the Cardiovascular Health Study. AB - OBJECTIVES: We sought to characterize the predictors of incident congestive heart failure (CHF), as determined by central adjudication, in a community-based elderly population. BACKGROUND: The elderly constitute a growing proportion of patients admitted to the hospital with CHF, and CHF is a leading source of morbidity and mortality in this group. Elderly patients differ from younger individuals diagnosed with CHF in terms of biologic characteristics. METHODS: We analyzed data from the Cardiovascular Health Study, a prospective population based study of 5,888 elderly people >65 years old (average 73 +/- 5, range 65 to 100) at four locations. Multiple laboratory measures of cardiovascular structure and function, blood chemistries and functional assessments were obtained. RESULTS: During an average follow-up of 5.5 years (median 6.3), 597 participants developed incident CHF (rate 19.3/1,000 person-years). The incidence of CHF increased progressively across age groups and was greater in men than in women. On multivariate analysis, other independent predictors included prevalent coronary heart disease, stroke or transient ischemic attack at baseline, diabetes, systolic blood pressure (BP), forced expiratory volume 1 s, creatinine >1.4 mg/dl, C-reactive protein, ankle-arm index <0.9, atrial fibrillation, electrocardiographic (ECG) left ventricular (LV) mass, ECG ST-T segment abnormality, internal carotid artery wall thickness and decreased LV systolic function. Population-attributable risk, determined from predictors of risk and prevalence, was relatively high for prevalent coronary heart disease (13.1%), systolic BP > or =140 mm Hg (12.8%) and a high level of C-reactive protein (9.7%), but was low for subnormal LV function (4.1%) and atrial fibrillation (2.2%). CONCLUSIONS: The incidence of CHF is high in the elderly and is related mainly to age, gender, clinical and subclinical coronary heart disease, systolic BP and inflammation. Despite the high relative risk of subnormal systolic LV function and atrial fibrillation, the actual population risk of these for CHF is small because of their relatively low prevalence in community-dwelling elderly people. PMID- 10807471 TI - The role of thoracic ultrasonography for evaluation of patients with decompensated chronic heart failure. AB - OBJECTIVES: This study examined the usefulness of thoracic ultrasonography for evaluation of fluid accumulation in patients with decompensated chronic heart failure (CHF) in comparison with physical signs, upright posteroanterior chest X ray and echocardiography. BACKGROUND: Decompensated CHF is frequently accompanied by pleural effusion, suggesting that pleural effusion is a useful marker for confirming the diagnosis of the uncontrolled stage of CHF. Thoracic ultrasonography seems to be adequate for this purpose. METHODS: Patients with uncontrolled CHF and an interpretable physical examination, chest X-ray, ultrasonogram for the heart and thorax and thoracic X-ray computed tomographic (CT) scan were enrolled in the study (n = 60). Patients free from thoracic and cardiovascular diseases served as a control (n = 22). Thoracic CT scan was used as the gold standard for the presence or absence of pleural effusion. Variables used to predict body fluid accumulation included the following: pulmonary rales, jugular venous distension or peripheral edema, roentgenographic evidence of pulmonary edema or pleural fluid, pericardial or pleural effusion on ultrasonographic study. RESULTS: The reported incidence of pleural effusion detected by thoracic ultrasonography was high (91%). The incidence of physical signs and roentgenographic signs of body fluid accumulation, however, was modest (56%) to low (33%). The best clinical variable for identifying patients with decompensated CHF was the detection of pleural fluid by thoracic ultrasonography (91% predictive accuracy). This variable also had high interobserver agreement (95% overall agreement, kappa = 0.70). There was only 41% to 65% predictive accuracy of other clinical variables, with 72% to 95% agreement (kappa = 0.400 0.848). CONCLUSIONS: Thoracic ultrasonography is a simple, sensitive and accurate method for the evaluation of body fluid accumulation in patients with decompensated CHF. This technique can be used to assist in making the diagnosis of decompensated CHF if other causes of pleural effusion have been clinically ruled out. PMID- 10807472 TI - Assessment of cardiac risk before nonvascular surgery: dobutamine stress echocardiography in 530 patients. AB - OBJECTIVE: This study evaluated the incremental value of dobutamine stress echocardiography (DSE) for assessment of cardiac risk before nonvascular surgery. BACKGROUND: Limited information exists regarding the preoperative assessment of cardiac risk in patients with known or suspected coronary artery disease who are to undergo nonvascular surgery. METHODS: All patients (303 men, 227 women) who underwent DSE before nonvascular surgery and did not sustain an intervening event (coronary revascularization or cardiac event) were studied. Clinical, electrocardiographic and rest and stress echocardiographic variables were evaluated to identify predictors of postoperative cardiac events. RESULTS: Events occurred in 6% of patients: 1 cardiac death and 31 nonfatal myocardial infarctions. All of these patients had inducible ischemia on DSE (sensitivity 100%, specificity 63%). Multivariate predictors of postoperative events in patients with ischemia were history of congestive heart failure (p = 0.006; odds ratio = 4.66; confidence interval 1.55 to 14.02) and ischemic threshold less than 60% of age-predicted maximal heart rate (p = 0.0001; odds ratio 7.002; confidence interval 2.79 to 17.61). Clinical variables of Eagle's index identified 21% of patients as low, 68% as intermediate and 11% as high risk preoperatively; the postoperative event rates were 3%, 6%, and 14%, respectively. Dobutamine stress echocardiography identified 60% of patients as low (no ischemia), 32% as intermediate (ischemic threshold 60% or more) and 8% as high risk (ischemic threshold < 60%); postoperative event rates were 0%, 9% and 43%, respectively. CONCLUSIONS: In this population of patients with known or suspected coronary artery disease evaluated before nonvascular surgery, DSE had incremental value over clinical, electrocardiographic and rest echocardiographic variables for identifying patients at low, intermediate and high risk for postoperative cardiac events. Ischemia occurring at less than 60% of age-predicted maximal heart rate identified patients at highest risk. PMID- 10807473 TI - Attenuated coronary flow reserve and vascular remodeling in patients with hypertension and left ventricular hypertrophy. AB - OBJECTIVES: The purpose of this study was to evaluate the association between hypertension and left ventricular hypertrophy (LVH) with both coronary vascular remodeling and endothelial function. BACKGROUND: The association between endothelial and nonendothelial coronary flow reserve with vascular remodeling in patients with hypertension and LVH is still unclear. METHODS: One hundred and eleven patients with normal or mildly diseased coronary arteries at angiography underwent intravascular ultrasound examination of the left anterior descending coronary artery. Patients were divided into three groups: group 1: n = 13, hypertensive patients with LVH; group 2: n = 30, hypertensive patients without LVH; group 3: n = 68, normotensive patients. Vessel and lumen area and atherosclerotic plaque area were evaluated. Vascular reactivity was examined using intracoronary adenosine and acetylcholine. RESULTS: Vessel area in group 1 (with LVH) was significantly (p < 0.01) greater than that in group 2 (without LVH), whereas, vessel area in both groups 1 and 3 was similar (12.8 +/- 0.8 mm2, 10.7 +/- 0.4 mm2 and 11.5 +/- 0.3 mm2). Coronary blood flow at baseline for patients in group 1 (with LVH) was significantly greater than it was for patients in groups 2 and 3 (81.1 +/- 9.9 ml/min, 56.5 +/- 6.2 ml/min and 48.1 +/- 3.2 ml/min, both p < 0.05). In comparison with groups 2 and 3, the response to both acetylcholine and adenosine was significantly impaired in patients with LVH. CONCLUSIONS: The current study demonstrates that hypertension with LVH is associated with both coronary vascular remodeling and attenuated endothelial and nonendothelial coronary flow reserve. PMID- 10807474 TI - Role of ischemia and infarction in late right ventricular dysfunction after atrial repair of transposition of the great arteries. AB - OBJECTIVES: This study was conducted to assess whether myocardial ischemia and/or infarction are involved in the pathogenesis of late right ventricular dysfunction in adult survivors of atrial baffle repair for transposition of the great arteries in infancy. BACKGROUND: The medium-term success of intraatrial baffle repair for transposition of the great arteries is good, with many patients surviving into adult life, but prognosis can be limited by progressive right ventricular dysfunction. We hypothesized that ongoing myocardial ischemia and/or infarction are important factors in the pathogenesis of this complication. Radionuclide techniques offer an opportunity to study both myocardial perfusion and concomitant ventricular wall motion. METHODS: Dipyridamole sestamibi single photon emission computed tomography followed by rest sestamibi single-photon emission computed tomography was used to assess right ventricular myocardial perfusion, wall motion, wall thickening and ejection fraction in 22 adolescents/young adults who had undergone atrial baffle repair for simple transposition of the great arteries at median 6.7 (range 0.5 to 54) months of age. The patients were aged 10 to 25 (median 15.5) years; 19 in New York Heart Association class I, 2 in class II and 1 in class III. All were in a regular cardiac rhythm during the studies. The right ventricular tomographic images were examined in three parallel and two orthogonal planes, analyzed in 12 segments. RESULTS: Perfusion defects were evident in all patients in at least one segment, in either the rest or stress images. Twelve patients (55%) demonstrated fixed defects only, nine (41%) had fixed and reversible defects and one (4.5%) had reversible defects only. Concomitant wall-thickening abnormalities occurred in 83% of segments with fixed perfusion defects, mirrored by a reduction in wall motion in 91% of segments analyzed. Right ventricular ejection fraction was correlated with age (R = 0.62; p = 0.002), and with wall-thickening abnormalities (R = 0.60; p < 0.005). CONCLUSIONS: Reversible and fixed perfusion defects with concordant regional wall motion abnormalities occur in the right (systemic) ventricle 10 to 20 years after Mustard repair for transposition of the great arteries; this may be important in the pathogenesis of late right ventricular dysfunction in this group. PMID- 10807475 TI - Increased expression of extracellular signal-regulated kinase and angiotensin converting enzyme in human atria during atrial fibrillation. AB - OBJECTIVES: The purpose of this study was to determine whether atrial expression of the extracellular signal-regulated kinases Erk1/Erk2 and of the angiotensin converting enzyme (ACE) is altered in patients with atrial fibrillation (AF). BACKGROUND: Recent studies have demonstrated that atrial fibrosis can provide a pathophysiologic substrate for AF. However, the molecular mechanisms responsible for the development of atrial fibrosis are unclear. METHODS: Atrial tissue samples of 43 patients undergoing open heart surgery were examined. Seventeen patients had chronic persistent AF (> or =6 months; CAF), 8 patients had paroxysmal AF (PAF) and 18 patients had no history of AF. Erk expression was analyzed at the mRNA (quantitative reverse transcription polymerase chain reaction), the protein (immunoblot techniques) and atrial tissue (immunohistochemistry) levels. Erk-activating kinases (MEK1/2) and ACE were analyzed by immunoblot techniques. RESULTS: Increased amounts of Erk2-mRNA were found in patients with CAF (75 +/- 20 U vs. sinus rhythm: 31 +/- 25 U; p < 0.05). Activated Erk1/Erk2 and MEK1/2 were increased to more than 150% in patients with AF compared to patients with sinus rhythm. No differences between CAF and PAF were found. The expression of ACE was three-fold increased during CAF. Amounts of activated Erk1/Erk2 were reduced in patients treated with ACE inhibitors. Patients with AF showed an increased expression of Erk1/Erk2 in interstitial cells and marked atrial fibrosis. CONCLUSIONS: An ACE-dependent increase in the amounts of activated Erk1/Erk2 in atrial interstitial cells may contribute as a molecular mechanism for the development of atrial fibrosis in patients with AF. These findings may have important impact on the treatment of AF. PMID- 10807476 TI - Ten-fold augmentation of endothelial uptake of vascular endothelial growth factor with ultrasound after systemic administration. AB - OBJECTIVES: In this study, the feasibility of delivering and enhancing the uptake of vascular endothelial growth factor (VEGF) into the intact endothelium by using ultrasound (US) facilitation was determined. BACKGROUND: A limitation of tissue targeted drug delivery is the need for direct arterial cannulation. We postulate a mechanism by which agents injected intravenously may be targeted to a tissue using US and ultrasonic contrast agents. METHODS: We used a rat model to test the ability of US and an ultrasonic contrast agent perflurocarbon exposed sonicated dextrose albumin (PESDA) to increase uptake of VEGF in the myocardium. Continuous wave Doppler US (0.6 W/cm2 at 1 MHz for 15 min) was applied to the chest wall overlying the myocardium during intravenous injection with either VEGF (100 microg/kg) alone or a combination of VEGF and PESDA (0.1%). Control rats had VEGF infused without US or PESDA. The VEGF uptake was measured quantitatively in the heart, lung, liver and kidneys by enzyme-linked immunosorbent assay (ng/g of tissue) and morphologically by fluorescence microscopy. RESULTS: There was an eight-fold increase in VEGF uptake in the heart by US alone (16.86 +/- 1.56 vs. 2.11 +/- 0.953 ng/g of tissue, p < 0.0001) and a 13-fold increase with US + PESDA (26.78 +/- 2.88 vs. 2.11 +/- 0.953 ng/g of tissue, p < 0.0001) compared with control rats. Fluorescence microscopy revealed deposition of VEGF in the endothelium of small intramyocardial arterioles. CONCLUSIONS: These results show a marked increase in endothelial VEGF uptake with US and US + PESDA. Thus, US may be used to augment endothelial VEGF uptake 10-fold to 13-fold. PMID- 10807477 TI - Mechanisms of atrial fibrillation: is a cure at hand? AB - The mechanisms of atrial fibrillation relate to the presence of random reentry involving multiple interatrial circuits. Triggers for development of atrial fibrillation include rapidly discharging atrial foci (mainly from pulmonary veins) or degeneration of atrial flutter or atrial tachycardia into fibrillation. Therapy for control of atrial fibrillation includes drugs, atrial pacing for those with sinus node dysfunction, or ablation of the atrioventricular junction. Therapeutic maneuvers for cure of atrial fibrillation include surgical or radiofrequency catheter induced linear lesions to reduce the atrial tissue and prevent the requisite number of reentrant wavelets. We need a much better understanding of basic mechanisms before a true cure is at hand. PMID- 10807478 TI - Full disclosure: the antidote to conflict of interest. PMID- 10807479 TI - President's page: convocation address. PMID- 10807480 TI - Heart rate-lowering drugs such as verapamil improve aerobic exercise performance in healthy elderly individuals: a new way to look at left ventricular diastolic function in the elderly. PMID- 10807481 TI - Anti-ischemic effect of enhanced external counterpulsation: where's the beef? PMID- 10807482 TI - A prospective, randomized, double-blind comparison of bilateral and right unilateral electroconvulsive therapy at different stimulus intensities. AB - BACKGROUND: Controversy persists about the use of right unilateral (RUL) and bilateral (BL) electroconvulsive therapy (ECT). While RUL ECT results in less severe short-term and long-term cognitive effects, there is concern that it is less efficacious than BL ECT. METHODS: In a double-blind study, 80 depressed patients were randomized to RULECT, with electrical dosages 50%, 150%, or 500% above the seizure threshold, or BL ECT, with an electrical dosage 150% above the threshold. Depression severity and cognitive functioning were assessed before, during, immediately after, and 2 months after ECT. Compared with baseline, responders had at least a 60% reduction in symptom scores 1 week after ECT, and were monitored for relapse for 1 year. RESULTS: High-dosage RUL and BL ECT were equivalent in response rate (65%) and approximately twice as effective as low dosage (35%) or moderate-dosage (30%) unilateral ECT. During the week after the randomized phase, BL ECT resulted in greater impairment than any dosage of unilateral ECT in several measures of anterograde and retrograde memory. Two months after ECT, retrograde amnestic deficits were greatest among patients treated with BL ECT. Thirty-three (53%) of the 62 patients who responded to ECT relapsed, without treatment group differences. The relapse rate was greater in patients who had not responded to adequate pharmacotherapy prior to ECT and who had more severe depressive symptoms after ECT. CONCLUSION: Right unilateral ECT at high dosage is as effective as a robust form of BL ECT, but produces less severe and persistent cognitive effects. PMID- 10807483 TI - Titrated moderately suprathreshold vs fixed high-dose right unilateral electroconvulsive therapy: acute antidepressant and cognitive effects. AB - BACKGROUND: The antidepressant and cognitive side effects of right unilateral (RUL) electroconvulsive therapy (ECT) are reported to depend on the magnitude of the electrical stimulus relative to the seizure threshold. The stimulus doses explored in previous clinical trials of RUL ECT have generally been limited to 1 to 2.5 times the convulsive threshold and the antidepressant efficacy has been low compared with bilateral (BL) ECT. The present study compares the antidepressant and cognitive side effects of 2 RUL dosing strategies: titrated moderately suprathreshold and fixed high dose. METHODS: Seventy-two adult patients with major depression were randomized to either titrated RUL ECT at 2.25 times initial seizure threshold (mean dose, 136 millicoulombes [mC]), or RUL ECT at a fixed dose of 403 mC. Primary outcome measures were antidepressant response and cognitive status 1 or 2 days after the course of ECT. RESULTS: The 2 treatment groups were comparable in demographic and clinical characteristics prior to ECT. Both groups received a mean of 5.7 sessions of RUL ECT. Patients receiving fixed-dose ECT were more likely to have an antidepressant response at the end of the protocol (n = 49 [67%]) compared with those receiving titrated dosing (n = 28 [39%]). Furthermore, the likelihood of both antidepressant response and cognitive deficits increased as stimulus dose increased relative to initial seizure threshold, up through 8 to 12 times the threshold. CONCLUSIONS: The antidepressant efficacy and cognitive side effects of RUL ECT are dependent on the magnitude of the stimulus dose relative to the seizure threshold, and a dose-response relationship extends through at least 12 times the seizure threshold. PMID- 10807484 TI - Electroconvulsive therapy requires higher dosage levels: Food and Drug Administration action is required. PMID- 10807485 TI - Decrease in thalamic volumes of pediatric patients with obsessive-compulsive disorder who are taking paroxetine. AB - BACKGROUND: Thalamic dysfunction has been implicated in obsessive-compulsive disorder (OCD). While OCD frequently has its onset during childhood, to our knowledge, no prior study has measured neuroanatomical changes in the thalamus of patients with OCD near the onset of illness, and before and after treatment. METHODS: Volumetric magnetic resonance imaging studies were conducted in 21 psychotropic drug-naive children, aged 8 to 17 years, with OCD and 21 case matched healthy comparison subjects. Magnetic resonance imaging studies were also conducted in 10 of the 21 patients with OCD after 12 weeks of monotherapy with the selective serotonin reuptake inhibitor, paroxetine hydrochloride. RESULTS: Thalamic volumes were significantly greater in treatment-naive patients with OCD than in controls but declined significantly after paroxetine monotherapy to levels comparable with those of controls. Decrease in thalamic volume in patients with OCD was associated with reduction in OCD symptom severity. CONCLUSIONS: Our findings provide new evidence of thalamic abnormalities in pediatric OCD and further suggest that paroxetine treatment may be paralleled by a reduction in thalamic volume. These reductions may, however, not be specific to paroxetine treatment and could be due to a more general treatment response, and/or spontaneous improvement in symptoms. Our findings are preliminary given the small sample size and our inability to measure discrete thalamic nuclei. PMID- 10807486 TI - A multicenter comparison of cognitive-behavioral therapy and interpersonal psychotherapy for bulimia nervosa. AB - BACKGROUND: Research suggests that cognitive-behavioral therapy (CBT) is the most effective psychotherapeutic treatment for bulimia nervosa. One exception was a study that suggested that interpersonal psychotherapy (IPT) might be as effective as CBT, although slower to achieve its effects. The present study is designed to repeat this important comparison. METHOD: Two hundred twenty patients meeting DSM III-R criteria for bulimia nervosa were allocated at random to 19 sessions of either CBT or IPT conducted over a 20-week period and evaluated for 1 year after treatment in a multisite study. RESULTS: Cognitive-behavioral therapy was significantly superior to IPT at the end of treatment in the percentage of participants recovered (29% [n=32] vs 6% [n=71), the percentage remitted (48% [n=53] vs 28% [n = 31]), and the percentage meeting community norms for eating attitudes and behaviors (41% [n=45] vs 27% [n=30]). For treatment completers, the percentage recovered was 45% (n= 29) for CBT and 8% (n= 5) for IPT. However, at follow-up, there were no significant differences between the 2 treatments: 26 (40%) CBT completers had recovered at follow-up compared with 17 (27%) IPT completers. CONCLUSIONS: Cognitive-behavioral therapy was significantly more rapid in engendering improvement in patients with bulimia nervosa than IPT. This suggests that CBT should be considered the preferred psychotherapeutic treatment for bulimia nervosa. PMID- 10807487 TI - Volumetric measure of the frontal and temporal lobe regions in schizophrenia: relationship to negative symptoms. AB - BACKGROUND: Previous research has provided evidence for brain abnormalities in schizophrenia, but their relationship to specific clinical symptoms and syndromes remains unclear. METHODS: With an all-male demographically similar sample of 53 schizophrenic patients and 29 normal control subjects, cerebral gray and white matter volumes (adjusted for intracranial volume and age were determined for regions in the prefrontal lobe and in the superficial and mesial temporal lobe using T1-weighted magnetic resonance imaging with 2.8-mm coronal slices. RESULTS: As a group, schizophrenic patients had wide-spread bilateral decrements in gray matter in the pre-frontal (7.4%) and temporal lobe regions (8.9%), but not in white matter in these regions. In the temporal lobe, gray matter reductions were found bilaterally in the superior temporal gyrus (6.0%), but not in the hippocampus and parahippocampus. While there were no overall group differences in white matter volumes, widespread decrements in prefrontal white matter in schizophrenic patients (n = 53) were related to higher levels of negative symptoms (partial r[49] = -0.42, P = .002), as measured by the Scale for the Assessment of Negative Symptoms. A post hoc analysis revealed that schizophrenic patients with high negative symptoms had generalized prefrontal white matter reductions (11.4%) that were most severe in the orbitofrontal subregion (15.1%). CONCLUSIONS: These results suggest that gray matter deficits may be a fairly common structural abnormality of schizophrenia, whereas reductions in prefrontal white matter may be associated with schizophrenic negative symptoms. PMID- 10807488 TI - A randomized, placebo-controlled 12-month trial of divalproex and lithium in treatment of outpatients with bipolar I disorder. Divalproex Maintenance Study Group. AB - BACKGROUND: Long-term outcomes are often poor in patients with bipolar disorder despite treatment; more effective treatments are needed to reduce recurrences and morbidity. This study compared the efficacy of divalproex, lithium, and placebo as prophylactic therapy. METHODS: A randomized, double-blind, parallel-group multicenter study of treatment outcomes was conducted over a 52-week maintenance period. Patients who met the recovery criteria within 3 months of the onset of an index manic episode (n = 372) were randomized to maintenance treatment with divalproex, lithium, or placebo in a 2:1:1 ratio. Psychotropic medications were discontinued before randomization, except for open-label divalproex or lithium, which were gradually tapered over the first 2 weeks of maintenance treatment. The primary outcome measure was time to recurrence of any mood episode. Secondary measures were time to a manic episode, time to a depressive episode, average change from baseline in Schedule for Affective Disorders and Schizophrenia-Change Version subscale scores for depression and mania, and Global Assessment of Function scores. RESULTS: The divalproex group did not differ significantly from the placebo group in time to any mood episode. Divalproex was superior to placebo in terms of lower rates of discontinuation for either a recurrent mood episode or depressive episode. Divalproex was superior to lithium in longer duration of successful prophylaxis in the study and less deterioration in depressive symptoms and Global Assessment Scale scores. CONCLUSIONS: The treatments did not differ significantly on time to recurrence of any mood episode during maintenance therapy. Patients treated with divalproex had better outcomes than those treated with placebo or lithium on several secondary outcome measures. PMID- 10807489 TI - Maintenance treatment in bipolar disorder. PMID- 10807490 TI - Major mental disorders and criminal violence in a Danish birth cohort. AB - BACKGROUND: This epidemiological investigation was designed to examine the relationships between each of the major mental disorders and criminal violence. Specifically, we assessed whether a significant relationship exists between violence and hospitalization for a major mental disorder, and whether this relationship differs for schizophrenia, affective psychoses, and organic brain syndromes. METHODS: Subjects were drawn from a birth cohort of all individuals born between January 1, 1944, and December 31, 1947, in Denmark (N = 358 180). Because of the existence of accurate and complete national registers, data were available on all arrests for violence and all hospitalizations for mental illness that occurred for individuals in this cohort through the age of 44 years. RESULTS: There was a significant positive relationship between the major mental disorders that led to hospitalization and criminal violence (odds ratios 2.0-8.8 for men and 3.9-23.2 for women). Persons hospitalized for a major mental disorder were responsible for a disproportionate percentage of violence committed by the members of the birth cohort. Men with organic psychoses and both men and women with schizophrenia were significantly more likely to be arrested for criminal violence than were persons who had never been hospitalized, even when controlling for demographic factors, substance abuse, and personality disorders. CONCLUSIONS: Individuals hospitalized for schizophrenia and men hospitalized with organic psychosis have higher rates of arrests for violence than those never hospitalized. This relationship cannot be fully explained by demographic factors or comorbid substance abuse. PMID- 10807491 TI - Evidence of the dual mechanisms of action of venlafaxine. AB - BACKGROUND: Indirect evidence suggests that the antidepressant venlafaxine hydrochloride selectively inhibits serotonin (5-HT) uptake at low doses, whereas at high doses, it inhibits both 5-HT and norepinephrine (NE) uptake. We hypothesized that, in vivo, both high and low doses would inhibit the 5-HT uptake of platelets but that the higher dose would differentially blunt the pressor response to tyramine, a marker for NE uptake. METHODS: Healthy male volunteers aged 18 to 45 years received either 75 mg or 375 mg of venlafaxine hydrochloride per day, the 5-HT uptake inhibitor sertraline hydrochloride (50 mg/d), or the NE uptake inhibitor maprotiline hydrochloride (150 mg/d) (n = 8 for each of 4 treatment groups). Changes in platelet 5-HT uptake and the pressor response to intravenous tyramine were assessed following the initial dose and after 1 and 2 weeks of drug administration. RESULTS: Platelet 5-HT uptake was inhibited by venlafaxine across the dose range and by sertraline but not maprotiline. Inhibition was competitive, related to increases in affinity and not related to capacity. Steady-state drug levels were associated with a 5-HT uptake inhibition of 87% or more in subjects taking venlafaxine or sertraline. The pressor response to tyramine differentially distinguished maprotiline from sertraline and the low dose of venlafaxine but not from the high dose of venlafaxine. CONCLUSION: This study provides the first in vivo evidence in healthy humans that both 5-HT uptake and NE uptake inhibition are mechanisms of action sequentially engaged by venlafaxine over its clinically relevant dose range. PMID- 10807492 TI - Testing for laterality differences in regional brain volumes. PMID- 10807493 TI - Nicotine dependence and withdrawal in cocaine-dependent patients. PMID- 10807494 TI - Age of onset and familial risk in major depression. PMID- 10807495 TI - Dexamethasone inhibits arteriolar leukocyte rolling and adhesion induced by tumor necrosis factor-alpha in vivo. AB - OBJECTIVE: Glucocorticoids are widely used in vasculitis syndromes, however, the protective mechanisms remain to be elucidated. In this study, we wanted to examine the effect of dexamethasone on leukocyte rolling and adhesion in arterioles in vivo. MATERIALS AND METHODS: NMRI mice were treated intrascrotally with tumor necrosis factor-alpha (TNF-alpha) for 3-4 h: Intravital microscopy was used to study leukocyte-endothelium interactions in arterioles in the cremaster muscle. RESULTS: It was found that TNF-alpha markedly increased the number of rolling and adherent leukocytes in the arterioles. Pretreatment (2 h) with dexamethasone significantly reduced TNF-alpha-induced leukocyte rolling and adhesion. In fact, 1 mg/kg of dexamethasone decreased leukocyte rolling and adhesion by 76% and 87%, respectively. CONCLUSIONS: These findings suggest that the beneficial effect of dexamethasone in vasculitis disorders may in part be attributable to an inhibitory impact on cytokine-induced leukocyte rolling and adhesion in arterioles. PMID- 10807496 TI - The potential role of the Arthus and Shwartzman reactions in the pathogenesis of pneumonic pasteurellosis. AB - Pneumonic pasteurellosis (PP) is an economically important disease in cattle, sheep, and goats. Pasteurella haemolytica is commonly isolated from the severe fibrinopurulent pneumonia that characterize this respiratory syndrome. During infection, the bacteria produce leukotoxin (LKT) and lipopolysaccharide (LPS), both potent inducers of inflammation. Nonetheless, it has also been demonstrated that an exacerbated host's inflammatory response is responsible for the severe lung damage. Despite research in this field, the pathogenesis of PP is still incomplete. Two classical models of acute inflammatory response induced in laboratory animals, the Arthus and Shwartzman reactions, could explain the pathogenesis of the severe lung lesions that characterize PP. PMID- 10807497 TI - Augmented prostaglandin E2 generation resulting from increased activities of cytosolic and secretory phospholipase A2 and induction of cyclooxygenase-2 in interleukin-1 beta-stimulated rat calvarial cells during the mineralizing phase. AB - OBJECTIVE AND DESIGN: To assess prostaglandin (PG) E2 production by osteoblasts during the mineralizing phase after interleukin (IL)-1beta stimulation, using an in vitro system of rat calvarial cells cultured for 21 days. METHODS: The cells, which reached confluence after 3 days, were designated day 0 cells. Culture was continued for a further 21 days after confluence. The cells on the 21st day of the culture were designated day 21 cells. RESULTS: The PGE2 concentration in the medium of the day 21 cells was increased 72 h after IL-1beta treatment, and reached a peak level approximately 1,400 times that of the day 0 cells 6 h after IL-1beta treatment. We examined the effects of IL-1beta on PGE2 production and changes in the relevant enzyme activities, and found that the activities of cytosolic phospholipase A2 (cPLA2), type II secretory PLA2 (sPLA2) and cyclooxygenase (COX)-2 in the day 21 cells were increased. Both selective COX-2 inhibitor and cPLA2 inhibitor abolished PGE2 generation, whereas an sPLA2 inhibitor partially inhibited it. Taken together, these results indicate that COX 2 and cPLA2 play pivotal roles and sPLA2 is involved in IL-1beta-stimulated PGE2 production by these cells. Furthermore, we found that IL-Ibeta treatment induced PGE synthase activity and this correlated well with PGE2 production. CONCLUSION: Augmented PGE2 production by mineralizing osteoblasts after IL-1beta treatment, and the involvement of IL-1beta-induced cPLA2, sPLA2, COX-2 and PGE synthase activities in this phenomenon were demonstrated. PMID- 10807498 TI - In vitro investigations with the histamine H1 receptor antagonist, epinastine (WAL 801 CL), on isolated human allergic effector cells. AB - OBJECTIVE AND DESIGN: Skin mast cells, basophils and eosinophils are effector cells of acute and subacute allergic responses due to their capacity to produce a large number of (pro)inflammatory mediators. Histamine H1 receptor antagonists, such as epinastine (WAL 801 CL), have been described to partially exert antiallergic and antiinflammatory effects both in vivo and in vitro in addition to their antihistaminergic properties. The aim of the present study was to investigate whether epinastine could influence the in vitro activation of isolated human skin mast cells, basophils and eosinophils induced by different secretagogues. METHODS: Cells were isolated from healthy women following plastic surgery and healthy blood donors, respectively. Mast cells were isolated by enzymatic digestion of the skin. Blood cells were isolated by gradient centrifugation and negative selection with magnetic beads. RESULTS: A wide range of concentrations of the drug (1 nmol/l to 100 micromol/l) did not significantly inhibit histamine release from basophils induced by immunologic (anti-IgE, concanavalin A, priming factors interleukin-3 and interleukin-5) and non immunologic (A23187, ionomycin, 12-o-tetradecanoyl-phorbol-13-acetate, C5a, formyl-methionyl-leucyl-phenylalanine) stimuli. Furthermore, the drug had no effect on A23187-induced release of eosinophil cationic protein from eosinophils. However, at a concentration >0.1 nmol/l, IgE-mediated LTC4 production from basophils was significantly suppressed. Histamine release from skin mast cells due to anti-IgE or A23187 was inhibited by epinastine in a dose-dependent fashion, whereas substance P-induced activation as well as stem cell factor priming were not. Epinastine did not inhibit isolated protein kinase C from rat brain. CONCLUSION: The results confirm previous in vivo and in vitro observations obtained from animal models that epinastine exerts antiallergic and antiinflammatory effects. Whether the observed effects are due to non specific membrane interactions or by influencing intracellular signal transduction elements has to be further elucidated. PMID- 10807499 TI - JTE-522 selectively inhibits cyclooxygenase-2-derived prostaglandin production in inflammatory tissues. AB - OBJECTIVE AND DESIGN: To investigate the effect of JTE-522, a selective cyclooxygenase (COX)-2 inhibitor, on prostaglandin (PG) production and COX expression in rats. SUBJECTS: Male rats (4-8 weeks old) were used for in vivo experiments, while for in vitro assay, rat peritoneal macrophages were used. TREATMENT: JTE-522 (1-100 mg/kg) and indomethacin (0.03-10 mg/kg) were administered orally. JTE-522 and reference compounds (0.01-10 microM) were subjected to COX expression. RESULTS: JTE-522 inhibited the development of carrageenin-induced paw edema and PGE2 production in inflammatory paws at a dose of 10 mg/kg. On the other hand, JTE-522 (1-100 mg/kg) did not affect A23187 stimulated thromboxane B2 release from whole blood or the PGE2 level in gastric mucosa. JTE-522 did not suppress lipopolysaccharide-induced COX-2 expression in peritoneal macrophages. CONCLUSION: These results indicate that JTE-522 selectively inhibits PG production mediated by COX-2 in inflammatory tissues. JTE 522 may thus represent a novel type of anti-inflammatory drug without adverse effects on the gastro-intestinal tract. PMID- 10807500 TI - The effects of contignasterol (IZP-94,005) on allergen-induced plasma protein exudation in the tracheobronchial airways of sensitized guinea-pigs in vivo. AB - OBJECTIVE: To examine contignasterol (marine-derived) on allergen-induced plasma exudation in the tracheobronchial airways. MATERIAL: English shorthair guinea pigs actively sensitized to ovalbumin. TREATMENT: 21 to 36 days after sensitization, contignasterol, budesonide or nedocromil and then ovalbumin (12 pmol) were superfused onto the tracheal mucosa (i. t.) of anaesthetized animals. METHODS: Tracer (125I-human serum albumin) was measured in tracheobronchial lavage. RESULTS: Thirty min after administration of contignasterol (100 and 200, but not 0.2 or 2.0 microg/kg), budesonide or nedocromil (200 microg/kg), the ovalbumin response (plasma exudate, microl) was significantly inhibited. The response was also inhibited 5 min after 200 microg/kg nedocromil but not after contignasterol or budesonide. CONCLUSIONS: Contignasterol was topically active and showed anti-inflammatory effects in the tracheobronchial airways in that it inhibited allergen-induced plasma protein exudation. PMID- 10807501 TI - Glucose phosphorylation capacity and glycolysis in the liver of arthritic rats. AB - OBJECTIVE AND DESIGN: Glycolysis and the glucose phosphorylation capacity of livers from arthritic rats were studied because alterations in these parameters are suggested by some studies. SUBJECTS: Arthritis was induced in male albino rats (Wistar; 180-220 g). TREATMENT: The animals were injected with 100 microl heat inactivated Mycobacterium tuberculosis suspended in mineral oil at a concentration of 0.5% (w/v). Animals showing lesions after 14 to 21 days were selected. METHODS: Glucose phosphorylation was measured in the high speed supernatant fraction of liver homogenates and glycolysis in the isolated perfused liver. RESULTS: The glucose concentration for half-maximal rates was reduced from 18.32+/-5.69 in normal to 9.84+/-3.15 mM in arthritic rats (p = 0.024). Vmax was increased from 8.77+/-0.27 in normal to 11.49+/-0.29 nmol min(-1) mg protein(-1) in arthritic rats (p = 0.001). Perfused livers from arthritic rats showed a 2.43 fold higher rate of glycolysis. CONCLUSIONS: Livers from arthritic rats present a higher glucose phosphorylation capacity. Possibly this phenomenon is caused by circulating inflammatory mediators produced during adjuvant-induced arthritis. PMID- 10807502 TI - A major metabolite of aceclofenac, 4'-hydroxy aceclofenac, suppresses the production of interstitial pro-collagenase/proMMP-1 and pro-stromelysin-1/proMMP 3 by human rheumatoid synovial cells. AB - OBJECTIVE AND DESIGN: We examined the effects of aceclofenac and its metabolites on the production of pro-collagenase-1/pro-matrix metalloproteinase-1 (proMMP-1), pro-gelatinase A/proMMP-2, pro-stromelysin-1/proMMP-3 and tissue inhibitor of metalloproteinases-1 (TIMP-1) by rheumatoid synovial cells. MATERIALS: Synovial cells were obtained from patients with rheumatoid arthritis. TREATMENT: Cultures of confluent cells were treated with interleukin-1beta (IL-1beta)(1 ng/ml) and/or test drugs (0.3-30 microM) for 48 h. METHODS: Production of proMMPs and TIMP-1 was monitored by Western blotting or gelatin zymography. Prostaglandin E2 (PGE2) was measured by an enzyme immunoassay. RESULTS: 4'-Hydroxy aceclofenac, a major metabolite of aceclofenac, down-regulated both basal and IL-1beta-induced production of proMMP-1 and proMMP-3 at a concentration sufficient to suppress PGE2 production without modulating proMMP-2 or TIMP-1, whereas aceclofenac itself had no marked effect on the production of proMMPs. CONCLUSIONS: Down-regulation of proMMP-1 and proMMP-3 production by 4'-hydroxy aceclofenac may contribute to the therapeutic effect of aceclofenac on rheumatoid arthritis and osteoarthritis. PMID- 10807503 TI - Innate and adaptive lymphoid cells in the human liver. AB - Because of its location and function, the liver is continuously exposed to a large antigenic load that includes pathogens, toxins, tumor cells and harmless dietary antigens. The range of local immune mechanisms required to cope with this diverse immunological challenge is now being appreciated. The liver has an "epithelial constitution" and contains large numbers of phagocytic cells, antigen presenting cells and lymphocytes and is a site for the production of cytokines, complement components and acute phase proteins. In this review, we focus on the hepatic lymphoid system, which is currently emerging as an important arm of the immune system in the liver for targeting pathogens as well as for the recognition of cells that are modified as a result of infection or tumor transformation. We show that this organ contains a heterogeneity of lymphoid cells with diverse recognition mechanisms and functions. There are conventional T lymphocytes that use clonotypic receptors to identify and respond to antigenic peptides presented in the context of polymorphic class I and class II major histocompatibilty complex (MHC) molecules. But these cells are outnumbered by lymphoid cells that recognize common structures using receptors with limited diversity. These mediators of innate immunity against infectious pathogens and malignant cells respond immediately to stimuli and function as a temporal (and perhaps evolutionary) bridge for the adaptive immune response. PMID- 10807504 TI - Local control of the immune response in the liver. AB - The physiological function of the liver--such as removal of pathogens and antigens from the blood, protein synthesis and metabolism--requires an immune response that is adapted to these tasks and is locally regulated. Pathogenic microorganisms must be efficiently eliminated while the large number of antigens derived from the gastrointestinal tract must be tolerized. From experimental observations it is evident that the liver favours the induction of tolerance rather than the induction of immunity. The liver probably not only is involved in transplantation tolerance but contributes as well to tolerance to orally ingested antigens (entering the liver with portal-venous blood) and to containment of systemic immune responses (antigen from the systemic circulation entering the liver with arterial blood). This review summarizes the experimental data that shed light on the molecular mechanisms and the cell populations of the liver involved in local immune regulation in the liver. Although hepatocytes constitute the major cell population of the liver, direct interaction of hepatocytes with leukocytes in the blood is unlikely. Sinusoidal endothelial cells, which line the hepatic sinusoids and separate hepatocytes from leukocytes in the sinusoidal lumen, and Kupffer cells, the resident macrophage population of the liver, can directly interact with passenger leukocytes. In the liver, clearance of antigen from the blood occurs mainly by sinusoidal endothelial cells through very efficient receptor-mediated endocytosis. Liver sinusoidal endothelial cells constitutively express all molecules necessary for antigen presentation (CD54, CD80, CD86, MHC class I and class II and CD40) and can function as antigen presenting cells for CD4+ and CD8+ T cells. Thus, these cells probably contribute to hepatic immune surveillance by activation of effector T cells. Antigen specific T-cell activation is influenced by the local microenvironment. This microenvironment is characterized by the physiological presence of bacterial constituents such as endotoxin and by the local release of immunosuppressive mediators such as interleukin-10, prostaglandin E2 and transforming growth factor beta. Different hepatic cell populations may contribute in different ways to tolerance induction in the liver. In vitro experiments revealed that naive T cells are activated by resident sinusoidal endothelial cells but do not differentiate into effector T cells. These T cells show a cytokine profile and a functional phenotype that is compatible with the induction of tolerance. Besides sinusoidal endothelial cells, other cell populations of the liver, such as dendritic cells, Kupffer cells and perhaps also hepatocytes, may contribute to tolerance induction by deletion of T cells through induction of apoptosis. PMID- 10807505 TI - The liver as a crucial organ in the first line of host defense: the roles of Kupffer cells, natural killer (NK) cells and NK1.1 Ag+ T cells in T helper 1 immune responses. AB - The liver remains a hematopoietic organ after birth and can produce all leukocyte lineages from resident hematopoietic stem cells. Hepatocytes produce acute phase proteins and complement in bacterial infections. Liver Kupffer cells are activated by various bacterial stimuli, including bacterial lipopolysaccharide (LPS) and bacterial superantigens, and produce interleukin (IL)-12. IL-12 and other monokines (IL- 18 etc.) produced by Kupffer cells activate liver natural killer (NK) cells and NK1.1 Ag+ T cells to produce interferon-gamma and thereby acquire cytotoxicity against tumors and microbe-infected cells. These liver leukocytes and the T helper 1 immune responses induced by them thus play a crucial role in the first line of defense against bacterial infections and hematogenous tumor metastases. However, if this defense system is inadequately activated, shock associated with multiple organ failure takes place. Activated liver NK1.1 Ag+ T cells and NK cells also cause hepatocyte injury. NK1.1 Ag+ T cells and another T-cell subset with an intermediate T-cell receptor, CD 122+CD8+ T cells, can develop independently of thymic epithelial cells. Liver NK cells and NK1.1 Ag+ T cells physiologically develop in situ from their precursors, presumably due to bacterial antigens brought from the intestine via the portal vein. NK cells activated by bacterial superantigens or LPS are also probably involved in the vascular endothelial injury in Kawasaki disease. PMID- 10807506 TI - The liver as a site of T-cell apoptosis: graveyard, or killing field? AB - The liver is a site at which apoptotic CD8+ cells accumulate during the clearance phase of peripheral immune responses. Normal mouse liver contains an unusual mixture of lymphocytes in which natural killer (NK) and NK-T cells are abundant and apoptotic T cells are present, and we interpret these cell populations as, respectively, agents and targets of an intrahepatic T-cell trapping and killing mechanism. In support of this idea, direct perfusion of activated lymphocyte populations through the normal liver results in the selective retention of activated CD8+ T cells. T cells trapped in this manner undergo apoptosis in the liver. This mechanism could explain the importance of the liver in oral tolerance, the phenomenon of tolerance induced by portal vein infusion of antigenic cells, the tolerance to allogeneic liver allografts, and the persistence of some liver pathogens including hepatitis C. PMID- 10807507 TI - Regulation and antimetastatic functions of liver-associated natural killer cells. AB - The liver is a complex organ composed of hepatic parenchymal cells and a variety of non-parenchymal cells that consist of endothelial cells, Kupffer cells, and several subsets of resident lymphocytes, including natural killer (NK), T, and NK1.1+/CD3+ (NK/T) cells. The regulation of these various lymphoid subpopulations and their relative contributions to antiviral, antitumor and pathogenic inflammatory responses in the liver remain topics of much interest. Studies from our laboratory have shown that various immune stimulants and cytokines can augment liver-associated NK activity at least partially through the mobilization of NK cells from the bone marrow to the liver. The mobilization process can be dependent on the induction of interferon (IFN)-gamma and/or tumor necrosis factor alpha and on very late activation antigen-4/vascular cell adhesion molecule-1 interaction. The induction of IFN-gamma by cytokines such as interleukin (IL)-12 also rapidly triggers the induction of chemokine genes in parenchymal cells that may contribute to the localization of NK and T cells. Both IL-2 and IL-12 trigger changes in the number and functions of liver-associated leukocyte subsets, and induce antimetastatic effects that are likely mediated through several direct and indirect mechanisms. The overall goal of these studies is to understand the interactions and functions of liver-associated NK1.1+ cells in the context of innate and adaptive immune responses to neoplasia. PMID- 10807508 TI - Life, activation and death of intrahepatic lymphocytes in chronic hepatitis C. AB - The healthy liver of adult humans has little or no lymphocyte component and the histological finding of intrahepatic lymphocytes (IHL) is evidence of liver pathology. In a liver injured by chronic hepatitis C, the most common chronic liver disease, most IHL are activated/pro-inflammatory cells, which are particularly enriched for effectors of innate immunity (natural killer (NK), natural T, and other NK-like T cells). IHL do not undergo clonal expansion in the liver but migrate from extrahepatic sites to the chronically infected liver, where they display effector function and subsequently die, suggesting that maintenance of the IHL pool depends on continuous lymphocyte migration. The cytotoxic and inflammatory functions of these IHL have three potential outcomes: 1) they could be helpful in clearing the virus (a rare case in hepatitis C virus (HCV) infection); 2) they could be useless and have no effect on the infection; or 3) they could be harmful, whereby overaggressive lymphocyte responses destroy the liver in a continuous and unsuccessful attempt to clear the virus. Unfortunately, we do not know as of yet which of these possibilities is the case and, therefore, a more complete picture of the intrahepatic immune response will be relevant to the development of new therapeutic strategies against HCV. Additionally and from a more general perspective, due to the availability of biopsied material and the high prevalence (approximately 3%) of HCV infection worldwide, studying the chronically inflamed liver of hepatitis C patients is an ideal model to investigate the poorly understood processes of lymphocyte trafficking, activation and death to non-lymphoid sites of chronic inflammation in man. PMID- 10807509 TI - Immunobiology of hepatitis C virus (HCV) infection: the role of CD4 T cells in HCV infection. AB - Hepatitis C virus (HCV) is yet another example of a pathogen that persists in the presence of a readily apparent immune response. As evidence for both humoral and cellular immune responsiveness is quite strong, our studies have begun to examine whether qualitative defects in CD4 T-cell responses to viral antigens may help to explain why HCV is not eliminated in the vast majority of infections. Direct evidence that CD4 T cells play a role in HCV persistence is lacking, but several observations are consistent with this possibility. Importantly, it does not exclude the role of antibody or killer T cells in the immunopathogenesis of HCV infection. In addition, we discuss the consequences of viral mutation and how naturally occurring variants in immunodominant viral epitopes can effectively suppress helper T-cell responses to wild type virus. PMID- 10807510 TI - Occult persistence and lymphotropism of hepadnaviral infection: insights from the woodchuck viral hepatitis model. AB - Hepatitis B virus (HBV) is a major human pathogen that causes chronic infection and life-threatening liver diseases in millions of individuals. While pathological and epidemiological consequences of clinically evident HBV infections are well recognized, there is no similar knowledge on an asymptomatic, silently progressing virus persistence. Contrary to previous opinion, current evidence indicates that a serologically undetectable (occult) HBV carriage is a common outcome of recovery from symptomatic illness and that scanty amounts of the virus are carried by apparently healthy individuals for years after resolution of hepatitis B despite the presence of presumably protective antiviral antibodies. Recent studies on this silent form of hepadnavirus carriage in an experimental woodchuck hepatitis virus (WHV) infection, which is considered to be the closest natural model of HBV disease, revealed that the life-long occult persistence of traces of pathogenic virus is an invariable consequence of recovery after hepadnaviral invasion and that this state always co-exists with a steady low-rate virus replication in both the liver and the lymphatic system. Importantly, this serologically concealed infection can be accompanied by development of hepatocellular carcinoma in convalescent animals and is transmittable from mothers to offspring as an asymptomatic, indefinitely long infection which involves the lymphatic system but not always the liver. This review focuses on the features of hepadnavirus occult persistence and its lymphotropism, and on what is currently understood about the contribution of the lymphatic system in maintaining hepadnavirus carriage based on insights provided by analysis of the woodchuck-WHV experimental system. PMID- 10807511 TI - CD8+ T-cell-mediated response to Listeria monocytogenes taken up in the liver and replicating within hepatocytes. AB - Like most other organisms that enter the bloodstream, the bulk of Listeria monocytogenes injected i.v. into mice is taken up by the liver. Listeriae not killed rapidly by infiltrating neutrophils are internalized by hepatocytes which constitute the principal site of intracellular replication in the liver. CD8+ T cells play a critical role in eliminating infected hepatocytes and resolving listerial infections of the liver; the specific mechanisms involved are not understood fully. Here, we review recent data implicating the cytolytic activities expressed by MHC class Ia- and class Ib-restricted CD8+ T cells in host resistance to Listeria. Evidence demonstrating the perforin- and/or Fas ligand-dependent induction of caspase activity and the resultant apoptosis of Listeria-infected hepatocytes is discussed. PMID- 10807512 TI - The role of intrahepatic lymphocytes in mediating protective immunity induced by attenuated Plasmodium berghei sporozoites. AB - Exposure to irradiated Plasmodium sporozoites (gamma-spz) results in protection against malaria. Like infectious spz, gamma-spz colonize hepatocytes to undergo maturation. Disruption of liver stage development prevents the generation of protection, which appears, therefore, to depend on liver stage antigens. Although some mechanisms of protection have been identified, they do not include a role for intrahepatic mononuclear cells (IHMC). We demonstrated that P. berghei gamma spz-immune murine IHMC adoptively transfer protection to naive recipients. Characterization of intrahepatic CD4+ T cells revealed an immediate, albeit transient, response to gamma-spz, while the response of CD8+ T cells is delayed until acquisition of protection. It is presumed that activated CD8+ T cells home to the liver to die; gamma-spz-induced CD8+CD45RB(lo)CD44(hi) T cells, however, persist in the liver, but not the spleen, during protracted protection. The association between CD8+CD45RB(lo)CD44(hi) T cells and protection has been verified using MHC class I and CD1 knockout mice and mice with disrupted liver stage parasites. Based on kinetic studies, we propose that interferon-gamma, presumably released by intrahepatic effector CD8+ T cells, mediates protection; the persistence of CD8+ T cells is, in turn, linked to Plasmodium antigen depots and cytokines released by CD4+ T cells and/or NK T cells. PMID- 10807513 TI - Physiological responses of extrathymic T cells in the liver. AB - The liver has been found to be one of the important hematopoietic organs even after birth. Namely, adult liver still comprises c-kit+ stem cells and gives rise to extrathymic T cells, natural killer (NK) cells, and even granulocytes. Extrathymic T cells generated in the liver of mice are intermediate T-cell receptor (TCR(int)) cells, including the NK1.1+TCR(int) (i.e. NKT cells) and NK1.1-TCR(int) subsets. Although extrathymic T cells are few in number in youth, they gradually increase in number with aging. Even in youth, the number and function of extrathymic T cells are elevated under conditions of stress, infection, malignancy, pregnancy, autoimmune disease, chronic graft-versus-host diseases, etc. Under these conditions, the mainstream of T-cell differentiation in the thymus, which produces conventional T cells, is rather suppressed. Since extrathymic T cells comprise self-reactive forbidden clones and mediate cytotoxicity against abnormal self-cells (e.g. malignant tumor cells, microbially infected hepatocytes, and regenerating hepatocytes), they are beneficial for the elimination of such cells. However, overactivation or continuous activation of extrathymic T cells might be harmful and responsible for the onset of autoimmune diseases. We finally propose the possibility that switching of the immune system from the thymus to the liver might be regulated by the autonomic nervous system as well as by cytokines. PMID- 10807514 TI - Innate defenses in the liver during Listeria infection. AB - The majority of pathogens entering the bloodstream are cleared by the liver. Listeria monocytogenes, an important natural pathogen of humans, is a useful tool for examining protective immune responses during systemic infections of mice. Innate immunity contributes to blood clearance and eventual sterilization of the liver subsequent to Listeria infections. Effector mechanisms expressed in the liver early after infections are orchestrated by complex interactions between resident populations, i.e. hepatocytes and Kupffer cells, with infiltrating monocytes, neutrophils, and natural killer cells. These interactions include cell to cell contact through adhesion molecules, as well as communication through secretion of cytokines and chemokines. The liver environment, as the interface between blood-borne pathogens and innate host defenses, is reviewed here. PMID- 10807515 TI - Cytokine regulation of liver injury and repair. AB - By comparing the hepatic responses to tumor necrosis factor (TNF)-alpha that occur during situations that promote liver injury (such as obesity or chronic exposure to ethanol) with those that occur after stimuli (such as partial hepatectomy) that lead to liver regeneration, it is apparent that hepatocytes are usually able to constrain noxious responses to TNF-alpha, such as the release of reactive oxygen from mitochondria. It appears that by promptly upregulating survival genes that regulate mitochondrial membrane permeability, hepatocytes are usually able to constrain noxious responses, including the release of mitochondrial-generated reactive oxygen species, that follow exposure to potentially toxic cytokines, such as TNF-alpha. Indeed, transient TNF-alpha mediated increases in ROS may even be exploited by hepatocytes to evoke a subsequent proliferative response. Thus, the healthy liver has well-developed defense mechanisms that permit hepatocytes to adapt to cytokine-initiated stress, protecting them from cytokine-mediated lethality. Nevertheless, these same cytokines may cause liver injury when hepatocytes have been pre-exposed to toxins (e.g. ethanol) that interfere with their usual protective responses. Furthermore, while transient adaptations to cytokine-initiated stress permit hepatocytes to survive and proliferate, persistence of these anti-apoptotic, adaptative responses (as occurs, for example, in fatty livers) may inadvertently enhance hepatocyte vulnerability to necrosis when the liver is confronted by secondary insults that promote mitochondrial membrane depolarization. PMID- 10807516 TI - Molecular pathogenesis of liver disease: an approach to hepatic inflammation, cirrhosis and liver transplant tolerance. AB - The hallmarks of chronic liver diseases are chronic inflammation, cellular damage, regeneration and fibrosis. An appreciation of intrahepatic molecular expression patterns in normal and diseased liver provides clues for understanding pathogenic pathways whilst studies of the structure and function of molecules implicated in liver disease provide insights into their potential as therapeutic targets. We have examined the expression, function, molecular structure and structure-function relationships of type IV dipeptidyl aminopeptidases. In particular, the roles of CD26/DPPIV in T-cell proliferation and chemotaxis and of fibroblast activation protein in human cirrhosis are discussed. We have investigated the pathogenesis of liver disease by characterising patterns of cytokine and growth factor expression in experimental and human cirrhosis. We have quite recently expanded this approach to use differential gene expression analyses to elucidate overall pathways of gene activation and suppression in human cirrhosis. In addition, our detailed molecular and cellular studies of the mechanisms of spontaneous liver transplant tolerance have generated novel insights into this process. This review touches on these diverse aspects of liver function and disease. PMID- 10807518 TI - Primary biliary cirrhosis: an orchestrated immune response against epithelial cells. AB - Primary biliary cirrhosis (PBC) is an organ-specific autoimmune disease that predominantly affects women and is characterized by chronic progressive destruction of small intrahepatic bile ducts with portal inflammation and ultimately fibrosis. The serologic hallmark of PBC is the presence of antibodies to mitochondria, especially to the E2 component of the pyruvate dehydrogenase complex. The mechanisms by which (and if) such antibodies produce liver tissue injury are unknown. However, the presence of these antibodies has allowed detailed immunological definition of the antigenic epitopes, the nature of reactive autoantibodies and the characterization of T-cell responses. Several mechanisms may now be proposed regarding the immune-mediated bile duct damage in PBC, including the possible role of T-cell-mediated cytotoxicity and intracellular interaction between the IgA class of antimitochondrial antibodies and mitochondrial autoantigens. There are major questions which remain unanswered, including, of course, etiology, but also the reasons for female predominance, the absence of PBC in children, the relative ineffectiveness of immunosuppressive drugs, and the specific role of mitochondrial antigens. The data so far provide suggestive evidence that PBC is a mucosal disease; this thesis provides a basis for discussion of etiology via the enterohepatic circulation of toxins and/or infection. PMID- 10807517 TI - Pathophysiological roles of interleukin-18 in inflammatory liver diseases. AB - Innate immune response to microbes sometimes determines the nature of the following specific immune response. Kupffer cells, a potent constituent of innate immunity, play a key role in developing the type 1 immune response by interleukin (IL)-12 production. Furthermore, Kupffer cells have the potential to induce liver injury by production of IL-18. Propionibacterium acnes-primed lipopolysaccharide (LPS)-challenged liver injury is the prototype of IL-18-induced tissue injury, in which IL-18 acts on natural killer cells to increase Fas ligand (FasL) that causes liver injury by induction of Fas-dependent hepatocyte apoptosis. LPS induces IL-18 secretion from Kupffer cells in a caspase-1-dependent manner. Indeed, caspase-1-deficient mice are resistant to P. acnes and LPS-induced liver injury. However, administration of soluble FasL induces acute liver injury in P. acnes-primed caspase-1-deficient mice but does not do so in IL-18-deficient mice, indicating that IL-18 release in a caspase-1-independent fashion is essential for this liver injury. Therefore, a positive feedback loop between FasL and IL-18 plays an important role in the pathogenesis of endotoxin-induced liver injury. PMID- 10807519 TI - The peculiar autoimmunity of primary biliary cirrhosis. AB - Autoantibodies to mitochondria (AMA, anti-M2) are a serologic hallmark of primary biliary cirrhosis (PBC). These react with three structurally and functionally related multienzymic complexes, the 2-oxoacid dehydrogenase complexes, but chiefly with the E2 subunit of pyruvate dehydrogenase complex (PDC-E2). Their very dose (95%) and specific association with PBC underpins the autoimmune concept of pathogenesis of that disease, notwithstanding several non-congruent features. Detailed studies, including structural analysis of epitopes, do not disclose how these autoantibodies originate. Their ubiquity in PBC has overshadowed the existence of a second set of relatively PBC-specific autoantibodies to nuclear antigens for which reactants have been cloned and characterized. These include centromeric proteins; proteins of the nuclear pore complex; nuclear dot proteins, which include Sp-100 and the promyelocytic leukemia antigen; and a recently identified autoantigen, SOX13. Certain of these reactants are DNA-binding proteins with transcriptional regulatory activity. Thus serum from individuals with the same clinical syndrome can have autoimmune reactivity to disparate mitochondrial and nuclear constituents in different cellular compartments. Antibody probing of phage displayed random peptide libraries, together with epitope scanning using overlapping sequential octameric peptides from the PDC-E2 sequence, showed that the discontinuous motifs MH, FV(E) and SYP contributed to a predicted conformational antibody epitope in the inner lipoyl domain of PDC-E2. PMID- 10807520 TI - Autoreactive responses to pyruvate dehydrogenase complex in the pathogenesis of primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a cholestatic liver disease characterised by immune-mediated destruction of the biliary epithelial cells (BEC) lining the intrahepatic bile ducts (non-suppurative destructive cholangitis (NSDC)). Autoantibody and autoreactive T-cell responses specific for the self-antigen pyruvate dehydrogenase complex (PDC) are almost ubiquitous in PBC patients, leading to the view that the disease has an autoimmune aetiology. Autoreactive responses in PBC appear to be directed at the E2 and at the E3-binding protein (E3BP) (protein X) components of PDC, with the dominant B-cell and T-cell epitopes in E2 (fewer data are available for E3BP) spanning the inner (of two) lipoic acid-binding domains. The causal link between the breakdown of self tolerance to PDC (particularly at the T-cell level) and the development of NSDC has been emphasised by the demonstration, in a murine model (experimental autoimmune cholangitis), that sensitisation with PDC of mammalian origin results in a breakdown of both B-cell and T-cell tolerance to murine PDC accompanied by the development of NSDC. An increasing understanding of the role played by PDC specific autoreactive T cells in the pathogenesis of PBC has led us to examine the role played by the target cells in PBC (BEC) in both the inducer and effector mechanisms responsible for PBC. PMID- 10807521 TI - Genetic susceptibilities for immune expression and liver cell injury in autoimmune hepatitis. AB - Genetic susceptibility to type 1 autoimmune hepatitis in white northern Europeans is related to female sex, HLA alleles encoding the six amino acid sequence LLEQKR at positions 67-72 of the DRB1 polypeptide, and CTLA-4 gene polymorphism. The principal HLA alleles associated with type 1 autoimmune hepatitis in Britain and North America are DRB1*0301 and DRB1*0401. In this model of susceptibility, lysine at position 71 of the expressed DR molecule is the critical amino acid. In Japan, Argentina and Mexico, susceptibility is linked to DRB1*0405 and DRB1*0404. These two alleles encode arginine at position 71 rather than lysine, but they share the motif LLEQ-R with DRB1*0401 and DRB1*0301. Thus, K or R at position 71 in the context of LLEQ-R may be critical for susceptibility. This "shared motif" or "epitope" may optimize T-cell recognition of autoantigen, and other alleles that encode lysine at DRbeta71 may also affect susceptibility and outcome, possibly by increasing the density of lysine or arginine 71 molecules on the surface of antigen-presenting cells. Since the DRB1*0301 allele is part of the extended ancestral 8.1 haplotype, it carries with it additional risk factors for autoimmunity, including TNFA*2 and C4A*Q0. Type 1 autoimmune hepatitis is a polygenic disorder and other yet undefined polymorphic genes may be non-specific immunoregulators. These additional MHC encoded genes and other non-MHC encoded genes may be important determinants of disease susceptibility and severity in type 1 autoimmune hepatitis. PMID- 10807522 TI - Unusual patterns of alloimmunity evoked by allogeneic liver parenchymal cells. AB - Despite the widely accepted view of the liver as an immunoprivileged tissue, purified allogeneic liver parenchymal cells delivered to the liver of a recipient mouse are highly antigenic. A functional transgenic model of hepatocyte transplantation in mice is used to explore host immune responses to allogeneic hepatocytes. Transplanted hepatocytes expressing human alpha-1-antitrypsin (hA1AT) are monitored for survival by the secretion of the transgene product hA1AT. Transplantation of transgenic hepatocytes into syngeneic or immunoincompetent severe combined immunodeficiency disease (SCID) mice results in indefinite hepatocellular allograft survival. However, transplantation of transgenic hepatocytes into allogeneic hosts results in rapid hepatocyte rejection. This rejection response is associated with prominent delayed type hypersensitivity responses to cellular alloantigen but minimal donor-reactive humoral immunity. Hepatocyte rejection is not controlled by host treatment with anti-CD4 mAb despite the ability of the same treatment regimen to produce indefinite survival of donor-matched heart allografts. Host immune responses to allogeneic hepatocytes utilize CD40L/CD40 but not CD28/B7 co-stimulation, unlike the activation of both of these systems in responses to other allografts. Furthermore, C57BL/6 mice which have been induced by anti-CD4 mAb or gallium nitrate treatment to accept heart allografts promptly reject donor-matched transgenic hepatocytes. Studies in reconstituted SCID, CD4 knockout (KO), and CD8 KO mice demonstrate that hepatocyte rejection can be initiated independently by either CD4+ T cells or CD8+ T cells, which again diverges from what has been observed for most other types of allografts. This may account for the relative resistance to immunoprotection for hepatocellular allografts with conventional immunosuppressive agents and to immunoregulatory states induced by other allografts. Three models of hepatocyte rejection are discussed. PMID- 10807523 TI - Immunological tolerance--the liver effect. AB - The developing immune system in the embryo is programmed to accept all its own tissues as self-products and not react against them. It has been the aim of transplantation research for the past 40 years to try and manipulate the developed immune system temporarily to a similar status to that in the embryo at the time of an organ graft, so that the organ graft is accepted but immunity generally is not interfered with. Such a state is known as immunological tolerance and can be produced in the laboratory by a number of special techniques, none of which is appropriate for clinical use in organ grafting. In pigs and rats, an allografted liver can sometimes survive permanently without any immunosuppressive treatment in immunologically mature animals. PMID- 10807524 TI - Function and regulation of sarcoplasmic reticulum Ca2+-ATPase: advances during the past decade and prospects for the coming decade. AB - In cardiac muscle, the contraction-relaxation cycle is tightly controlled by the regulated release and uptake of intracellular Ca2+ between sarcoplasmic reticulum and cytoplasm. A major protein controlling Ca2+ cycling is Ca2+-ATPase (SERCA2a) located in the sarcoplasmic reticulum membrane. The function of SERCA2a protein is regulated by the phosphorylatable protein, phospholamban. Phosphorylation of phospholamban releases its inhibitory effect on SERCA2a through direct molecular interaction. Recently, mice whose SERCA2a function is increased (overexpression of the gene) or lost (knock out) were developed. These mice demonstrated that SERCA2a pump levels are a major determinant of cardiac muscle contractility and relaxation. These studies open the prospect that the overexpression of SERCA2a can correct cardiac dysfunction seen in heart failure. Advances in knowledge concerning the function and gene regulation of SERCA2a are discussed in this review. PMID- 10807525 TI - Takayasu arteritis: insights into immunopathology. AB - Takayasu arteritis is an acute and sometimes chronic form of vasculitis involving the aorta, its main branches and pulmonary arteries. Although its etiology is still unknown, immunopathologic analyses revealed that the infiltrating cells mainly consisted of gammadelta T-cells as well as alphabeta T-cells and NK cells. The infiltrating gammadelta T-cells, cytotoxic T-lymphocytes (CTLs), and natural killer (NK) cells directly injured the vascular cells by releasing a cytolytic factor, perforin. Expression of heat-shock protein (HSP)-65 as well as human leukocyte antigen (HLA) class I and II was enhanced in Takayasu arteritis lesions, supporting the pathogenic role of gammadelta T-cells and alphabeta T cells. T-cell receptor (TCR) alphabeta gene usage by the infiltrating cells was restricted, strongly suggesting that a specific antigen was targeted. TCR gammadelta gene usage by the infiltrating cells was also restricted. Furthermore, it has been reported that a strong association with a specific haplotype of major histocompatibility complex (MHC) class I chain-related (MIC), MICA gene with Takayasu arteritis, suggesting that the HLA-linked gene susceptible to the disease is mapped near the MICA gene. This also supports a pathogenic role of gammadelta T-cells in Takayasu arteritis because gammadelta T-cells were shown to recognize MICA molecule, which can be stress-induced. These findings suggest that unknown stress, such as infection, may trigger the autoimmune process of inflammation involved in Takayasu arteritis. PMID- 10807526 TI - Serum levels of vascular endothelial growth factor and transforming growth factor beta1 in patients with atrial fibrillation undergoing defibrillation therapy. AB - We have previously reported that pulsatile mechanical stretch in vitro induced rapid secretion of vascular endothelial growth factor (VEGF) by cultured cardiac myocytes and that the stretch-induced secretion of VEGF was mainly mediated by secretion of transforming growth factor (TGF)-beta1 by cardiac myocytes in an autocrine fashion. To investigate whether tachycardia-induced mechanical overload increases serum levels of VEGF and TGF-beta1, we investigated the serum levels of VEGF and TGF-beta1 in patients with atrial fibrillation undergoing defibrillation therapy. The serum VEGF level before defibrillation was significantly increased in 13 out of 20 patients (89.48 +/- 16.09 pg/ml [mean +/- SE]). After defibrillation, the serum VEGF level in these 13 patients significantly (p = 0.019) decreased (65.04 +/- 16.61 pg/ml [mean +/- SE]), although it increased slightly in one patient. The serum TGF-beta1 level before defibrillation therapy (13.01 +/- 1.97 pg/ml [mean +/- SE]) in these 12 patients also decreased after defibrillation therapy (11.47 +/- 2.06 pg/ml [mean +/- SE]). The changes in serum VEGF level significantly correlated with those in the serum TGF-beta1 level in these 12 patients (r = 0.73, p < 0.05, n = 12). Our data suggest that tachyarrhythmia-induced mechanical overload can increase the serum VEGF level, which can be a useful clinical marker for relative myocardial oxygen shortage in such patients. PMID- 10807527 TI - Modulation of the sympathovagal balance in drug refractory dilated cardiomyopathy, treated with permanent atrioventricular sequential pacing. AB - The aim of this study was to assess the long term efficacy of DDD pacing mode in selected patients with idiopathic dilated cardiomyopathy (IDCM) and drug refractory heart failure. The patients were evaluated according to the long term alteration of the sympathovagal balance (SVB). Patients with IDCM were considered eligible for DDD pacing if during temporary VDD pacing a 15% or more increase in the resting cardiac output was demonstrated. From the 29 patients studied, finally 20 patients (15M, 5F, 69 +/- 10 years) fulfilled the aforementioned criterion and therefore were considered candidates for permanent DDD pacing (NYHA class: 3.5 +/- 0.3, Ejection fraction: 27 +/- 7%, Resting cardiac index (CI) 2.6 +/- 0.4 l/min). The ECG of the patients demonstrated LBBB in 13, RBBB in 4 and RBBB + LAH in 3, with a PR interval of 232 +/- 28 ms and QRS duration of 138 +/- 15 ms. The pacemaker was programmed at 40-150 bpm, and AV delay of 105 +/- 20 ms. The lower heart rate programmed, in conjunction with the heart failure state of these patients, was responsible for essentially continuous atrial tracking, ventricular pacing. We evaluated the SVB in the pre- and post-implant periods (3rd and 6th month), using the hourly power spectral analysis (PSA) of heart rate variability during 24 hour Holter monitoring. As SVB we considered the ratio: low (0.04-0.15 Hz) to high frequency (0.15-0.40 Hz). We compared the SVB (LF/HF) during the day and night time for the pre- and post-implant periods. Post-pacing, the NYHA class was significantly improved (2.9 +/- 0.2 and 2.7 +/- 0.3 the 3rd and 6th month respectively). The mean heart rate was 78 +/- 8 bpm in the 3rd and 80 +/- 7 bpm in the 6th month postoperatively, which was lower than the 84 +/- 9 bpm preoperatively, but this difference did not reach statistical significance. During the night time the LF/HF decreased from 1.45 +/- 0.2 (LF: 7.19 +/- 0.43, HF: 4.95 +/- 0.54) in the pre-implant period to 0.9 +/- 0.09 (p < 0.001) (LF: 6.96 +/- 0.63, HF: 7.73 +/- 0.48) in the 3rd month. No further changes were observed in the 6th month (0.82 +/- 0.05, p = NS) (LF: 6.83 +/- 0.51, HF: 8.53 +/ 0.86) compared to the 3rd month. During the day time the LF/HF decreased from 1.5 +/- 0.5 (LF: 7.87 +/- 0.67, HF: 5.24 +/- 0.32) to 1.43 +/- 0.6 (p = NS) (LF: 7.34 +/- 0.71, HF: 5.24 +/- 0.42) in the 3rd month and to 1.41 +/- 0.09 in the 6th month (p = NS) (LF: 7.51 +/- 0.74, HF: 5.36 +/- 0.63). Comparing the LF/HF of day and the night time period, while in the pre-implant period there was no significant difference (1.5 +/- 0.5 vs 1.45 +/- 0.2, p = NS), the difference became significant in the 3rd (1.43 +/- 0.6 vs 0.9 +/- 0.09, p < 0.001) and 6th month (1.41 +/- 0.09 vs 0.82 +/- 0.05, p < 0.001). In conclusion, DDD pacing with individualized AV delay as an adjunct therapy could be a valuable method in selected patients with IDCM and drug refractory heart failure. DDD pacing improves the SVB over the long term. This improvement is attributed to sympathetic activity withdrawal and is more pronounced during night and less during day time. PMID- 10807528 TI - C-reactive protein as a prognostic marker in lymphocytic myocarditis. AB - The prognosis of patients with lymphocytic myocarditis (LM) is poor with the combined endpoint of death or transplant in the Myocarditis Treatment trial being 56% at 5 years. Physicians often have difficulty determining the prognosis in an individual patient. Patients with LM may have ongoing myocardial inflammation. We evaluated the ability of a serum marker of inflammation to predict prognosis in patients with LM. Serum concentrations of C-reactive protein (CRP) were measured in patients with LM. Thiry-one patients with clinical and histologic evidence of LM were evaluated. Patients with coronary artery disease, and idiopathic dilated and secondary cardiomyopathies were excluded. Overall mortality and morbidity from congestive heart failure was assessed at 28 days. The mean plasma CRP concentrations in the five patients who died within the 28-day follow-up period were significantly higher than in those patients who survived (17.4 +/- 5.6 vs 5.9 +/- 3.3 mg/ml, p < 0.05). The CRP concentration was positively correlated with plasma levels of lactic dehydrogenase and the New York Heart Association functional class. Routine measurement of CRP may be a useful tool for determining the prognosis in patients with LM. PMID- 10807529 TI - Ambulatory blood pressure monitoring after successful repair of coarctation of the aorta at mid-term follow-up. AB - Variations of systolic and diastolic blood pressure in patients with normotension at rest after successful surgical repair of coarctation of the aorta were examined using 24 hour ambulatory monitoring at mid-term follow-up. Ambulatory blood pressure monitoring, m-mode measurements of left ventricle and transmitral Doppler spectrals in 18 patients aged 7.6 +/- 4.5 years after 9 months to 6.1 years (2.5 +/- 1.9 years) following operation were compared with the findings in 18 matched controls. Patients had significantly higher mean systolic blood pressures (24 hours: 115 +/- 10 mmHg, awake: 119 +/- 11 mmHg and asleep: 106 +/- 8 mmHg) than controls (24 hours: 108 +/- 6 mmHg, awake: 112 +/- 7 mmHg and asleep: 101 +/- 7 mmHg) (p = 0.04, 0.03 and 0.03, respectively). Patients had also more systolic blood pressure readings above the 95th percentile for age (24 hours: 28 +/- 20%, awake: 39 +/- 27% and asleep: 12 +/- 14%) than controls (24 hours: 10 +/- 9%, awake: 14 +/- 13% and asleep: 1 +/- 4%) (p = 0.03, 0.002 and 0.007, respectively). No significant difference was found in diastolic blood pressure profiles. There were no significant differences in left ventricular m mode measurements and diastolic function parameters. Left ventricular mass index was significantly increased in patients (81.7 +/- 28.7 g/m2) compared with controls (64.5 +/- 20.9 g/m2) (p = 0.03). Operation age and post-surgical period did not affect ambulatory blood pressure profiles at mid-term follow-up. Patients who are normotensive at rest after successful surgical repair of coarctation of the aorta show higher systolic blood pressure profiles than healthy children with ambulatory blood pressure monitoring at mid-term follow-up. Ambulatory blood pressure monitoring in patients operated on for coarctation of the aorta despite their good clinical status is useful to detect and monitor subtle abnormalities of blood pressure. PMID- 10807530 TI - Identification of regulated genes in rat heart after myocardial infarction by means of differential mRNA display. AB - In order to testify the hypothesis that unknown mechanisms are involved in the process of cardiac remodeling after myocardial infarction (MI), we employed differential display reverse transcription-polymerase chain reaction (DDRT-PCR) as our primary inspection tool. An animal model of MI was established by ligation of the left anterior descending coronary artery (LAD) in rat. Fifty upregulated candidate cDNA fragments were obtained in the right ventricle (RV) of the heart six weeks after MI. Eight cDNA fragments isolated from DD denaturing gel were extracted and reamplified, cloned into pCR II vector and sequenced. A Genbank search of these clones showed that three of them have a high homology with known genes not previously associated with cardiac remodeling, i.e., mouse interleukin 4 receptor gene, rat ferritin mRNA, and T-cell receptor beta chain V beta 5. The remaining clones have no similarity to known sequences. These data suggest that certain genes which were not previously being associated with cardiac hypertrophy are turned on during the process of cardiac remodeling after MI. PMID- 10807531 TI - Advantage of recombinant technology for the identification of cardiac myosin epitope of severe autoimmune myocarditis in Lewis rats. AB - Whole cardiac myosin induced experimental autoimmune myocarditis (EAM) in animals. The identification of the epitope causing EAM is expected to elucidate the mechanism leading to the onset of this autoimmune disease. Until now such studies have been done mostly with synthetic oligo-peptides. We employed recombinant technology to produce the immunogenic fragment of the self-cardiac myosin heavy chain (CMHC) for EAM in Lewis rats, and successfully induced severe myocarditis with short recombinant peptide fragment CMHC residues 1107 to 1186. The immunogenicity was completely abolished from the fragment with further excision of 12 amino acids from residues 1131 to 1143. This recombinant technology clearly has advantages in the ease of manipulation and the purity for the creation of immunogenic epitopes. PMID- 10807532 TI - Therapy with the nonpeptide endothelin receptor antagonist 97-139 in a murine model of congestive heart failure: reduction of cardiac mass and myofiber hypertrophy. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor. This peptide exerts numerous effects on the heart, including regulation of cardiomyocyte growth during hypertrophy. The effects of the structurally novel, nonpeptide, ET-1-selective, competitive antagonist (ETA) 97-139 were investigated in mice with congestive heart failure (CHF) and myocardial hypertrophy. Morphological and microscopical analyses were conducted on day 56 after viral inoculation following 28 day treatment with 99-139. Eight week-old DBA2 mice were intraperitoneally inoculated with encephalomyocarditis virus at a dose of 500 pfu/mouse. The 30 mice were divided into two groups--an ETA treated group and an untreated group. Heart weight (HW) in the infected group was significantly (p < 0.05) increased compared to that in the uninfected group. HW and the HW/body weight (BW) ratio were significantly (p < 0.05) reduced in the ETA treated group compared with the untreated group (HW; 127.7 +/- 6.2 mg vs 144.3 +/- 4.2 mg, HW/BW; 4.9 +/- 0.9 x 10(-3) vs 5.4 +/- 0.5 x 10(-3)). Myofiber diameter in the ETA treated group was significantly reduced compared with the untreated group (12.1 +/- 1.5 microm vs 14.3 +/- 1.9 microm). These results suggest the ET-1 receptor antagonist 97-139 has an effect on the reduction of cardiac mass and myofiber hypertrophy, and that 97-139 may be a useful agent for CHF due to viral myocarditis. PMID- 10807533 TI - Local antithrombotic therapy using a novel porous balloon catheter. AB - The efficacy of local treatment of thrombosis with low-dose antithrombotic drugs (heparin: 30 U/kg, or argatroban: 0.02 mg/kg) was investigated using a novel porous balloon catheter. This novel balloon catheter can deliver drug into arterial walls without causing vascular trauma. Thrombus formation was significantly inhibited in balloon-injured and locally-treated iliac arteries compared with control balloon-injured arteries in 12 dogs. In the systemic high dose delivery group (ten times as high as the low dose), thrombus formation in injured arteries was significantly less than that of controls in 7 dogs. Low-dose systemic delivery was not effective at inhibiting this thrombus formation. Thus, local treatment with an antithrombotic drug using this novel porous balloon catheter can prevent thrombosis without influencing systemic coagulability. PMID- 10807534 TI - Transient left ventricular aneurysm and hypertrophy accompanied by polymorphic ventricular tachycardia in a patient suspected of acute myocarditis. AB - A 75-year-old woman presented with recurrent ventricular tachycardia (VT) compatible with torsades de pointes (TdP) based on sinus bradycardia and QT prolongation. Previously she had received pirmenol, at a serum concentration within therapeutic range, for her paroxysmal atrial fibrillation. Emergent cardiac catheterization identified a ventricular aneurysm of the anteroapical and inferior wall along with angiographically normal coronary arteries. A right ventricular endomyocardial biopsy revealed postmyocarditic change. The left ventricular contraction improved after 5 weeks of conservative treatment. A follow-up echocardiogram revealed transient thickening of partial left ventricular wall consistent with the segment of the aneurysm. Several months later, almost all abnormal findings had improved except for sustained deep negative T waves in precordial leads. Acute myocarditis was primarily suspected as the cause of her clinical presentation. PMID- 10807535 TI - X-chromosome inactivation in the human trophoblast of early pregnancy. AB - To investigate X-chromosome inactivation (XCI) in human trophoblasts during early pregnancy, trophoblast genomic DNA was extracted and analyzed for a Bst XI restriction endonuclease site polymorphism in the X-linked phosphoglycerate kinase gene, after digestion with methylation-sensitive Hpa II (control samples were digested instead with Afa I). Six villous trophoblast DNA samples were informative for the polymorphism (ie, heterozygous) and were derived from women homozygous for the polymorphism. These samples were then evaluated for XCI. In five of the six samples with Hpa II predigestion, the sizes of the two heterozygous band peaks differed; maternal X-chromosome (X(M))-derived alleles showed smaller peak sizes than paternal X-chromosome (X(P))-derived alleles, but the differences varied in degree. In samples obtained by microdissection from formalin-fixed, paraffin-embedded tissues (30 samples from different anchoring villi, and 38 samples from different branch villi), monoclonal band patterns of X(P)-derived alleles were observed more frequently than those of X(M)-derived alleles, but almost half of the samples showed polyclonal patterns. Our results suggest that a skewing of XCI exists in the human trophoblast; however, the degree of nonrandomness due to predominant X(P) inactivation appears to be restricted. It is probable that transcription of the X inactivation center (XIC) begins earlier in mice than in humans. PMID- 10807536 TI - Molecular analysis of alpha-thalassemia in Nepal: correlation with malaria endemicity. AB - Thalassemia is a prevalent hereditary disorder characterized by impaired synthesis of globin chains. It has been suggested that the high frequency of thalassemia might reflect heterozygote advantage due to reduced susceptibility to malaria. In Nepal, malaria has often occurred in places below the altitude of 1,200m. We carried out a microepidemiological study on thalassemia in two neighboring populations in Nepal, the Danuwar and the Tamang. Settlements of the Danuwar are located below the limit of the malarial zone (1,200m in altitude), whereas those of the Tamang are found in malaria-free uplands. Three heterozygotes for hemoglobin E (HbE) were observed in the Danuwars. We detected one type (-alpha3.71) of alpha+-thalassemia that involves a deletion of 3.7kb, leading to a loss of one of two alpha-globin genes, in the Danuwars, at a high gene frequency of 63%, while the gene frequency in the Tamangs was only 5%. Analysis of the alpha-globin gene cluster revealed that four different haplotypes were associated with the type of alpha+-thalassemia in the Danuwars. Nucleotide sequences of the D-loop region in the mitochondrial DNA of the two populations indicated a similar nucleotide diversity in each population. The fixation index, FST, representing the degree of genetic differentiation estimated from mitochondrial DNA diversities (FST, 0.05), was smaller than that obtained from the gene frequencies of alpha-thalassemia (FST, 0.55). If we assume neutral molecular evolution in the D-loop region of mitochondrial DNA, these results suggest that the high frequency of alpha+-thalassemia may be due to biological adaptation to the malarial environment rather than to events such as a bottleneck. PMID- 10807537 TI - Identification of Rad51 alteration in patients with bilateral breast cancer. AB - The human Rad51 gene, HsRAD51, is a homolog of RecA of Escherichia coli and functions in recombination and DNA repair. BRCA1 and BRCA2 proteins form a complex with Rad51, and these genes are thought to participate in a common DNA damage response pathway associated with the activation of homologous recombination and double-strand break repair. Additionally, we have shown that the pattern of northern blot analysis of the RadS gene is closely similar to those of the BRCA1 and BRCA2 genes. It is therefore possible that alterations of the Rad51 gene may be involved in the development of hereditary breast cancer. To investigate this possibility, we screened Japanese patients with hereditary breast cancer for Rad51 mutations and found a single alteration in exon 6. This was determined to be present in the germline in two patients with bilateral breast cancer, one with synchronous bilateral breast cancer and the other with synchronous bilateral multiple breast cancer. In both patients, blood DNAs showed a G-to-A transition in the second nucleotide of codon 150, which results in the substitution of glutamine for arginine. As this alteration was not present in any patients with breast or colon cancer examined, we assume that this missense alteration is likely to be a disease-causing mutation. PMID- 10807538 TI - The 4G/5G polymorphism of the plasminogen activator inhibitor-1 gene is associated with severe preeclampsia. AB - Preeclampsia is associated with thrombosis of the intervillous or spiral artery. A deletion/insertion polymorphism (4G or 5G) in the promoter of the plasminogen activator inhibitor type 1 (PAI-1) gene is suggested to be involved in regulating the synthesis of the inhibitor, 4G allele, being associated with the enhanced gene expression and plasma PAI-1 levels. We assessed the association between preeclampsia and the 4G/5G polymorphism of the PAI-1 gene in 115 preeclamptic patients, 210 pregnant controls, and 298 healthy volunteer controls. The frequency of the homozygotes for the 4G allele was significantly higher in the patients than in the control pregnant women (P = 0.04) or in the healthy volunteers (P = 0.02). The 4G allele frequency was also significantly higher in the patients than in the control group of pregnant women (P = 0.03) and in the healthy volunteers (P = 0.02). These results suggest that the presence of the 4G/4G genotype of the PAI-1 gene is one of the risk factors for preeclampsia. PMID- 10807539 TI - A Japanese propositus with D-- phenotype characterized by the deletion of both the RHCE gene and D1S80 locus situated in chromosome 1p and the existence of a new CE-D-CE hybrid gene. AB - In a family study of a Japanese propositus with the D-- phenotype, the serological data of her D-- phenotype and those of her parents were discrepant. Gene analysis of the propositus showed a gross deletion of the RHCE gene and a new rearrangement of RHCE to yield the CE-D-CE hybrid. It was demonstrated that the hybrid CE-D-CE gene consisted of exon 1 from the RHCE gene, followed by exons 3 to 7 from the RHD gene and exons 8 to 10 from the RHCE gene. However, whether or not exon 2 of the RHD or the RHCE gene was contained in the CE-D-CE gene remained unclear. Moreover, spacer analysis between both RH genes and the family study suggested that the D-- gene complex from the paternal and maternal sides consisted of only the CE-D-CE hybrid gene and a single RHD gene, respectively. For the purpose of confirming the parent-child relationship, a paternity test using DNA fingerprint and polymerase chain reaction (PCR) analysis at the D1S80 locus were performed. DNA fingerprints with two kinds of DNA minisatellite probes (33.15 and 33.6) confirmed that the parent-child relationship in the D-- propositus was compatible. However, in the present case, at the D1S80 locus, the PCR product derived from the mother was lacking, thereby negating a parent-child relationship. It is probable that the RH genes and D1S80 locus exist in close proximity, because they are situated in chromosomes 1p 34.3-36.1 and 1p 36.1 36.3, respectively. These data suggested that at the stage of gametogenesis, both the RHCE gene and the D1S80 locus from the maternal side may have been deleted, thereby producing the D-- gene complex. PMID- 10807541 TI - Novel mutations of the FANCG gene causing alternative splicing in Japanese Fanconi anemia. AB - Fanconi anemia (FA), an autosomal recessive disorder characterized by a progressive pancytopenia associated with congenital anomalies and high predisposition to malignancies, is a genetically and clinically heterogeneous disease. At least eight complementation groups (FA-A to FA-H) have been identified. Previously, we studied mutations of the FANCA gene, responsible for FA-A, and found pathogenic mutations in 12 of 15 unclassified Japanese FA patients. Here, we further studied an additional 5 FA patients for sequence alterations of the FANCA gene and found pathogenic mutations in 2 of them. We further analyzed mutations of the FANCC and FANCG genes, responsible for FA-C and FA-G, respectively, in the remaining 6 FA patients. Although there was no alterations in the FANCC gene in these 6 patients, two novel mutations of the FANCG gene, causing aberrant RNA splicing, were detected in 2 FA patients. One was a base substitution from G to C of the invariant GT dinucleotides at the splice donor site of intron 3, resulting in the skipping of exon 3, as well as the skipping of exons 3 and 4. The other was a base substitution from C to T in exon 8, creating a nonsense codon (Q356X). This mutation resulted in the exclusion of a sequence of 18 nucleotides containing the mutation from the mRNA, without affecting the splicing potential of either the authentic or the cryptic splice donor site. Collectively, 14 of the 20 unclassified Japanese FA patients belong to the FA-A group, 2 belong to the FA-G group, and none belongs to the FA C group. PMID- 10807540 TI - Co-segregation of elevated LDL with a novel mutation (D92K) of the LDL receptor in a kindred with multiple lipoprotein abnormalities. AB - Factors predisposing to the phenotypic features of familial combined hyperlipidemia have not been clearly defined. In the course of investigating familial coronary artery disease in Utah, we identified a three-generation family in which multiple members were affected with type IIa hyperlipoproteinemia (HLP IIa), type IIb hyperlipoproteinemia (HLP IIb), or type IV hyperlipoproteinemia (HLP IV). Because several family members had relatively severe low-density lipoprotein (LDL) cholesterol elevation, in order to dissect the possible contribution to the plasma lipoprotein abnormalities in this pedigree, we identified a novel point mutation in the low-density lipoprotein receptor (LDLR) gene, a G-to-A transition at nucleotide position 337 in exon 4. This change substituted lysine for glutamic acid at codon 92 (D92K) of the LDL receptor. By means of mutant allele-specific amplification we determined that the mutation co segregated with elevated cholesterol and LDL cholesterol in the plasma of family members with HLP IIa and HLP IIb, but not with the elevated plasma triglycerides seen in HLP IIb and HLP IV patients. Thus, in families with apparent familial combined hyperlipidemia, a defective LDLR allele and other genetic or environmental factors that elevate plasma triglycerides may account for the multiple lipid phenotypes observed in this kindred. PMID- 10807542 TI - Genetic alteration of the DCX gene in Japanese patients with subcortical laminar heterotopia or isolated lissencephaly sequence. AB - We examined mutations of the doublecortin (DCX) gene, which is responsible for X linked subcortical laminar heterotopia (SCLH) and lissencephaly, in eight unrelated Japanese patients, four with SCLH and four with isolated lissencephaly sequence (ILS). Polymerase chain reaction (PCR) disclosed a deletion of part of the DCX gene in one male ILS patient. Single-strand conformational polymorphism analysis and subsequent sequence analysis were carried out in the remaining seven patients. One male ILS patient had a nonsense mutation in exon V, which would result in premature termination of the gene product. One female SCLH patient had a missense mutation in exon IV. Our results indicate that in the Japanese, as has been seen elsewhere, abnormality of the DCX gene is the common cause of SCLH and ILS. PMID- 10807543 TI - Human amniotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver. AB - As human amniotic epithelial tissue is formed on about the eighth day after fertilization, human amniotic epithelial cells (hAEC) may have multipotency to differentiate into various organs, such as brain, heart, or liver. In this study, we showed evidence of the synthesis and excretion of albumin by hAEC, by immunostaining and enzyme-linked immunoassay. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analyses revealed the expression of albumin mRNA and protein, respectively. In addition, hAEC also demonstrated immunoreactivity to genetic markers of liver lineage, such as human serum albumin and alpha-fetoprotein. Transplanted hAEC to Scid mouse liver showed positive immunoreactivity to albumin and alpha-fetoprotein. Genetically modified cells containing the beta-galactosidase (LacZ) gene (AxCALacZ) were integrated in liver parenchyma. Human polymorphic gene analysis in Scid mouse liver after the implantation of hAEC showed that these Scid mouse livers obviously contained this human-specific gene until day 7 after the cell transplantation. As hAEC do not cause any acute rejection by allotransplantation, we conclude that hAEC may be useful as a transgene carrier to treat patients with inherited liver diseases. PMID- 10807544 TI - p24/ING1-ALT1 and p47/ING1-ALT2, distinct alternative transcripts of p33/ING1. AB - p33/ING1s (the growth inhibitor ING1 and candidate tumor suppressor ING1) are key players in the suppressive pathways for human tumorigenesis. We analyzed their complete transcripts, primary structures, and expression. The results led us to discover two novel and related alternatively spliced transcripts encoding p24/ING1-ALT1 and p47/ING1-ALT2. They share C-terminal residues with the candidate tumor suppressors p33/ING1. The candidate tumor suppressors p33/ING1 and p24/ING1-ALT1 were coexpressed in a variety of fetal and adult human tissues, but p47/ING1-ALT2 was minimally expressed. PMID- 10807545 TI - Twenty single nucleotide polymorphisms (SNPs) and their allelic frequencies in four genes that are responsible for familial long QT syndrome in the Japanese population. AB - We report here 20 single nucleotide polymorphisms (SNPs), including 10 novel ones, and their allelic frequencies detected in four genes that are known to be responsible for familial long QT syndrome in the Japanese population; 7 polymorphisms are in the KCNQ1 gene, 6 in the KCNH2 gene, 5 in the SCN5A gene, and 2 in the KCNE1 gene. These data will be of use for genetic association studies of acquired cardiac arrhythmias. PMID- 10807546 TI - Disparity between association and linkage analysis for HNF1A G319S in type 2 diabetes in Canadian Oji-Cree. AB - In parallel experiments designed to find the genetic determinants of type 2 diabetes in Oji-Cree, we identified several linked chromosomal regions, using genomic scanning, in addition to a private diabetes-associated mutation, namely HNF1A G319S, using candidate gene sequencing. The genome scan did not identify the region harboring HNF1A as being linked with diabetes. Also, the HNF1A mutation, when used directly in sib-pair linkage analysis, was not linked with diabetes. However, HNF1A G319S was very strongly associated with diabetes, predicted the clinical severity of diabetes, and performed well as a diagnostic predictive test for diabetes in the Oji-Cree. Despite the failure of linkage analysis to identify HNF1A as a determinant of type 2 diabetes, we feel justified in interpreting that G319S has a very important pathogenic role in Oji-Cree diabetes, based upon the highly suggestive association studies. The probable etiologic heterogeneity of type 2 diabetes in the Oji-Cree created a situation in which association analysis was much more sensitive to detect a relationship between HNF1A S319 and diabetes than was linkage analysis. The effectiveness of linkage analysis will probably be limited in study samples that have an even greater complexity of genetic background and/or disease etiology. Thus, the absence of linkage does not always mean that a genomic variant is not an important determinant of a complex disease. Furthermore, our experience confirms the value of using several complementary strategies to identify susceptibility genes for a complex disease. PMID- 10807547 TI - Molecular cloning and expression analysis of the human DA41 gene and its mapping to chromosome 9q21.2-q21.3. AB - DA41 was previously identified as one of the DAN-binding proteins, via a yeast based two-hybrid screening strategy. In the present study, we cloned a human homolog of DA41 cDNA. Structural analysis revealed that human DA41 cDNA consisted of 2,861 nucleotides in length and encoded a protein of 589 amino acids, with a predicted molecular mass of 62.4kDa. Human DA41 exhibited an 86% amino acid sequence identity to rat DA41, indicating the evolutionarily conserved structure and function of DA41. A database search for DA41-related protein(s) identified mouse PLIC-1, PLIC-2, frog XDRP1, and yeast DSK2. DA41 and each DA41-related protein contain a ubiquitin-like domain in their amino-terminal regions. DA41 was expressed ubiquitously in adult human tissues, with relatively higher levels in pituitary gland, adrenal gland, kidney, thymus, and placenta. Fluorescence in situ hybridization (FISH) revealed that DA41 was mapped to human chromosome 9q21.2-q21.3, a position overlapping the candidate tumor suppressor locus for bladder cancer. PMID- 10807548 TI - cDNA cloning, tissue expression, and chromosome mapping of human homolog of SOX18. AB - The SRY (sex-determining region Y) gene encodes a transcription factor characterized by a DNA-binding motif termed the HMG (high mobility group) domain. The SOX (Sry-box) genes comprise a large family related by homology to the HMG box region. We isolated a cDNA clone with an open reading frame encoding a putative protein of 384 amino acids, which shared 83% identity to the mouse Sox18 protein. Northern blot analysis revealed that a 1.9-kb band of human SOX18 messenger RNAs was predominantly expressed in heart, although weak signals were seen in brain, liver, testis, and leukocyte. By polymerase chain reaction (PCR) based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid panel, the gene was mapped to the chromosome 20q13.33 region. PMID- 10807549 TI - Trichloroethylene and cancer: epidemiologic evidence AB - adopt a traditional review of the mutagenicity data on TCE and its metabolites but instead raise several issues regarding the interpretation of mutagenicity and genetic toxicity tests in shedding light on whether these processes are key events in tumor initiation. As discussed in the U.S. EPA proposed cancer guidelines, a salient question is whether TCE or its metabolites interacts directly with and mutates DNA to bring about changes in gene expression or whether DNA mutation is achieved through some other process. The Moore and Harrington-Brock article examines this question. Bull ((italic)10(/italic)), Lash et al. ((italic)11(/italic)), and Green ((italic)12(/italic)) present the experimental support for several modes of action for tumor development in rodents. These articles discuss a number of hypotheses including the influence on tumor development from mutagenesis, cytotoxicity, cell proliferation, *(subscript)2u(/subscript)-globin, peroxisome proliferation, oxidative stress, receptor binding, and perturbation of cell-signaling pathways. Quantitative dose response issues important to the statistical modeling of both noncarcinogenic and carcinogenic effects are discussed in articles by Fisher ((italic)5(/italic)), Bois ((italic)13(/italic)), Clewell et al. ((italic)14(/italic)), Bois ((italic)15(/italic)), Boyes et al. ((italic)16(/italic)), Barton and Clewell ((italic)17(/italic)), Chen ((italic)18(/italic)), and Rhomberg ((italic)19(/italic)). Because pharmacokinetic data are available for the TCE assessment, dose metrics other than applied dose may be evaluated in benchmark and other dose-response analyses. Fisher's article ((italic)5(/italic)) describes modeling liver concentration of TCE and its oxidative metabolites, while Clewell et al. ((italic)14(/italic)) model plasma concentrations of the oxidative metabolites and flux through the kidney for metabolites of the glutathione (italic)S(/italic)-transferase pathway. Both models are scaled from mice or rats to humans and provide estimates of human equivalent doses simulating inhalation and oral exposure routes. Parameters from these models have been further subjected to an uncertainty analysis in the articles by Bois ((italic)13,15(/italic)). The application of Baysian statistical methods is increasingly used for updating estimates of pharmacokinetic model parameters. Moreover, these analyses can provide an additional set of dosimetric estimates that in some instances are very different from those obtained with the original model. These findings make the risk assessor's job more complex. Boyes and others ((italic)16(/italic)) test whether Haber's Law or a dose metric that integrates time and concentration best describes neurologic effects with high-level TCE exposure. Barton and Clewell ((italic)17(/italic)) examine both experimental and pharmacokinetically derived dose metrics in their analysis of neurologic and systemic organ toxicity seen in the rodent studies. The article further applies benchmark dose methodology to the quantitative analysis of these effects. The U.S. EPA proposed cancer guidelines recommend that dose-response modeling be carried out in two parts: analysis of the curve shape within the range of the data and extrapolation below the observable data. Application of a biologically based model is preferred for evaluating the dose-response relationship for carcinogenic effects. Such an approach is described in the article by Chen ((italic)18(/italic)), which explores the relationship between TCE and two of its oxidative metabolites, dichloroacetic acid and trichloroacetic acid, under the hypothesis that these chemicals induce liver tumors in mice through promotion of preexisting initiated cells. Unfortunately, the data ne PMID- 10807550 TI - Trichloroethylene and cancer: epidemiologic evidence. AB - Trichloroethylene is an organic chemical that has been used in dry cleaning, for metal degreasing, and as a solvent for oils and resins. It has been shown to cause liver and kidney cancer in experimental animals. This article reviews over 80 published papers and letters on the cancer epidemiology of people exposed to trichloroethylene. Evidence of excess cancer incidence among occupational cohorts with the most rigorous exposure assessment is found for kidney cancer (relative risk [RR] = 1.7, 95% confidence interval [CI] 1.1-2.7), liver cancer (RR = 1.9, 95% CI(1.0-3.4), and non-Hodgkin's lymphoma (RR = 1.5, 95% CI 0.9-2.3) as well as for cervical cancer, Hodgkin's disease, and multiple myeloma. However, since few studies isolate trichloroethylene exposure, results are likely confounded by exposure to other solvents and other risk factors. Although we believe that solvent exposure causes cancer in humans and that trichloroethylene likely is one of the active agents, we recommend further study to better specify the specific agents that confer this risk and to estimate the magnitude of that risk. PMID- 10807551 TI - Metabolism of trichloroethylene. AB - A major focus in the study of metabolism and disposition of trichloroethylene (TCE) is to identify metabolites that can be used reliably to assess flux through the various pathways of TCE metabolism and to identify those metabolites that are causally associated with toxic responses. Another important issue involves delineation of sex- and species-dependent differences in biotransformation pathways. Defining these differences can play an important role in the utility of laboratory animal data for understanding the pharmacokinetics and pharmacodynamics of TCE in humans. Sex-, species-, and strain-dependent differences in absorption and distribution of TCE may play some role in explaining differences in metabolism and susceptibility to toxicity from TCE exposure. The majority of differences in susceptibility, however, are likely due to sex-, species-, and strain-dependent differences in activities of the various enzymes that can metabolize TCE and its subsequent metabolites. An additional factor that plays a role in human health risk assessment for TCE is the high degree of variability in the activity of certain enzymes. TCE undergoes metabolism by two major pathways, cytochrome P450 (P450)-dependent oxidation and conjugation with glutathione (GSH). Key P450-derived metabolites of TCE that have been associated with specific target organs, such as the liver and lungs, include chloral hydrate, trichloroacetate, and dichloroacetate. Metabolites derived from the GSH conjugate of TCE, in contrast, have been associated with the kidney as a target organ. Specifically, metabolism of the cysteine conjugate of TCE by the cysteine conjugate ss-lyase generates a reactive metabolite that is nephrotoxic and may be nephrocarcinogenic. Although the P450 pathway is a higher activity and higher affinity pathway than the GSH conjugation pathway, one should not automatically conclude that the latter pathway is only important at very high doses. A synthesis of this information is then presented to assess how experimental data, from either animals or from (italic)in vitro (/italic)studies, can be extrapolated to humans for risk assessment. (italic)Key words(/italic): conjugate beta-lyase, cysteine glutathione, cytochrome P450, glutathione (italic)S(/italic)-transferases, metabolism, sex dependence, species dependence, tissue dependence, trichloroethylene. PMID- 10807553 TI - Mutagenicity of trichloroethylene and its metabolites: implications for the risk assessment of trichloroethylene. AB - This article addresses the evidence that trichloroethylene (TCE) or its metabolites might mediate tumor formation via a mutagenic mode of action. We review and draw conclusions from the published mutagenicity and genotoxicity information for TCE and its metabolites, chloral hydrate (CH), dichloroacetic acid (DCA), trichloroacetic acid (TCA), trichloroethanol, S-(1, 2-dichlorovinyl) l-cysteine (DCVC), and S-(1, 2-dichlorovinyl) glutathione (DCVG). The new U.S. Environmental Protection Agency proposed Cancer Risk Assessment Guidelines provide for an assessment of the key events involved in the development of specific tumors. Consistent with this thinking, we provide a new and general strategy for interpreting genotoxicity data that goes beyond a simple determination that the chemical is or is not genotoxic. For TCE, we conclude that the weight of the evidence argues that chemically induced mutation is unlikely to be a key event in the induction of human tumors that might be caused by TCE itself (as the parent compound) and its metabolites, CH, DCA, and TCA. This conclusion derives primarily from the fact that these chemicals require very high doses to be genotoxic. There is not enough information to draw any conclusions for trichloroethanol and the two trichloroethylene conjugates, DCVC and DCVG. There is some evidence that DCVC is a more potent mutagen than CH, DCA, or TCA. Unfortunately, definitive conclusions as to whether TCE will induce tumors in humans via a mutagenic mode of action cannot be drawn from the available information. More research, including the development and use of new techniques, is required before it is possible to make a definitive assessment as to whether chemically induced mutation is a key event in any human tumors resulting from exposure to TCE. PMID- 10807552 TI - Human variability and susceptibility to trichloroethylene. AB - Although humans vary in their response to chemicals, comprehensive measures of susceptibility have generally not been incorporated into human risk assessment. The U.S. EPA dose-response-based risk assessments for cancer and the RfD/RfC (reference dose-reference concentration) approach for noncancer risk assessments are assumed to protect vulnerable human subgroups. However, these approaches generally rely on default assumptions and do not consider the specific biological basis for potential susceptibility to a given toxicant. In an effort to focus more explicitly on this issue, this article addresses biological factors that may affect human variability and susceptibility to trichloroethylene (TCE), a widely used halogenated industrial solvent. In response to Executive Order 13045, which requires federal agencies to make protection of children a high priority in implementing their policies and to take special risks to children into account when developing standards, this article examines factors that may affect risk of exposure to TCE in children. The influence of genetics, sex, altered health state, coexposure to alcohol, and enzyme induction on TCE toxicity are also examined. PMID- 10807556 TI - Pulmonary toxicity and carcinogenicity of trichloroethylene: species differences and modes of action. AB - Trichloroethylene (TCE) is both acutely toxic and carcinogenic to the mouse lung following exposure by inhalation. In contrast, it is not carcinogenic in the rat lung and is markedly less toxic following acute exposure. Toxicity to the mouse lung is confined almost exclusively to the nonciliated Clara cell and is characterized by vacuolation and increases in cell replication. Chloral, a metabolite of TCE that accumulates in Clara cells and has been shown to be the cause of the toxicity, also causes aneuploidy in some test systems. Cytotoxicity, increased cell division, and aneuploidy are known risk factors in the development of cancer and provide a plausible mode of action for TCE as a mouse lung carcinogen. All acute and chronic effects of TCE on the mouse lung are believed to be a direct consequence of high cytochrome P450 activity and impaired metabolism of chloral in Clara cells. Comparisons between species suggest that the ability of the human lung to metabolize TCE is approximately 600-fold less than that in the mouse. In addition, the human lung differs markedly from the mouse lung in the number and morphology of its Clara cells. Thus, the large quantitative differences between the metabolic capacity of the mouse lung and the human lung, together with the species differences in the number and morphology of lung Clara cells, suggest that the risks to humans are minimal and that other tumor sites should take precedent over the lung when assessing the potential risks to humans exposed to TCE. PMID- 10807554 TI - Modes of action of trichloroethylene for kidney tumorigenesis. AB - This article focuses on the various models for kidney toxicity due to trichloroethylene (TCE) and its glutathione-dependent metabolites, in particular S-(1,2-dichlorovinyl)-l-cysteine. Areas of controversy regarding the relative importance of metabolic pathways, species differences in toxic responses, rates of generation of reactive metabolites, and dose-dependent phenomena are highlighted. The first section briefly reviews information on the incidence and risk factors of kidney cancer in the general U.S. population. Epidemiological data on incidence of kidney cancer in male workers exposed occupationally to TCE are also summarized. This is contrasted with cancer bioassay data from laboratory animals, that highlights sex and species differences and, consequently, the difficulties in making risk assessments for humans based on animal data. The major section of the article considers proposed modes of action for TCE or its metabolites in kidney, including peroxisome proliferation, alpha(2u)-globulin nephropathy, genotoxicity, and acute and chronic toxicity mechanisms. The latter comprise oxidative stress, alterations in calcium ion homeostasis, mitochondrial dysfunction, protein alkylation, cellular repair processes, and alterations in gene expression and cell proliferation. Finally, the status of risk assessment for TCE based on the kidneys as a target organ and remaining questions and research needs are discussed. PMID- 10807557 TI - Physiologically based pharmacokinetic models for trichloroethylene and its oxidative metabolites. AB - Trichloroethylene (TCE) pharmacokinetics have been studied in experimental animals and humans for over 30 years. Compartmental and physiologically based pharmacokinetic (PBPK) models have been developed for the uptake, distribution, and metabolism of TCE and the production, distribution, metabolism, and elimination of P450-mediated metabolites of TCE. TCE is readily taken up into systemic circulation by oral and inhalation routes of exposure and is rapidly metabolized by the hepatic P450 system and to a much lesser degree, by direct conjugation with glutathione. Recent PBPK models for TCE and its metabolites have focused on the major metabolic pathway for metabolism of TCE (P450-mediated metabolic pathway). This article briefly reviews selected published compartmental and PBPK models for TCE. Trichloroacetic acid (TCA) is considered a principle metabolite responsible for TCE-induced liver cancer in mice. Liver cancer in mice was considered a critical effect by the U.S. Environmental Protection Agency for deriving the current maximum contaminant level for TCE in water. In the literature both whole blood and plasma measurements of TCA are reported in mice and humans. To reduce confusion about disparately measured and model-predicted levels of TCA in plasma and whole blood, model-predicted outcomes are compared for first-generation (plasma) and second-generation (whole blood) PBPK models published by Fisher and colleagues. Qualitatively, animals and humans metabolize TCE in a similar fashion, producing the same metabolites. Quantitatively, PBPK models for TCE and its metabolites are important tools for providing dosimetry comparisons between experimental animals and humans. TCE PBPK models can be used today to aid in crafting scientifically sound public health decisions for TCE. PMID- 10807555 TI - Mode of action of liver tumor induction by trichloroethylene and its metabolites, trichloroacetate and dichloroacetate. AB - Trichloroethylene (TCE) induces liver cancer in mice but not in rats. Three metabolites of TCE may contribute--chloral hydrate (CH), dichloroacetate (DCA), and trichloroacetate (TCA). CH and TCA appear capable of only inducing liver tumors in mice, but DCA is active in rats as well. The concentrations of TCA in blood required to induce liver cancer approach the mM range. Concentrations of DCA in blood associated with carcinogenesis are in the sub-microM range. The carcinogenic activity of CH is largely dependent on its conversion to TCA and/or DCA. TCA is a peroxisome proliferator in the same dose range that induces liver cancer. Mice with targeted disruptions of the peroxisome proliferator-activated receptor alpha (PPAR-alpha) are insensitive to the liver cancer-inducing properties of other peroxisome proliferators. Human cells do not display the responses associated with PPAR-alpha that are observed in rodents. This may be attributed to lower levels of expressed PPAR-alpha in human liver. DCA treatment produces liver tumors with a different phenotype than TCA. Its tumorigenic effects are closely associated with differential effects on cell replication rates in tumors, normal hepatocytes, and suppression of apoptosis. Growth of DCA induced tumors has been shown to arrest after cessation of treatment. The DCA and TCA adequately account for the hepatocarcinogenic responses to TCE. Low-level exposure to TCE is not likely to induce liver cancer in humans. Higher exposures to TCE could affect sensitive populations. Sensitivity could be based on different metabolic capacities for TCE or its metabolites or result from certain chronic diseases that have a genetic basis. PMID- 10807558 TI - Statistical analysis of Fisher et al. PBPK model of trichloroethylene kinetics. AB - Two physiologically based pharmacokinetic models for trichloroethylene (TCE) in mice and humans were calibrated with new toxicokinetic data sets. Calibration is an important step in model development, essential to a legitimate use of models for research or regulatory purposes. A Bayesian statistical framework was used to combine prior information about the model parameters with the data likelihood to yield posterior parameter distributions. For mice, these distributions represent uncertainty. For humans, the use of a population statistical model yielded estimates of both variability and uncertainty in human toxicokinetics of TCE. After adjustment of the models by Markov chain Monte Carlo sampling, the mouse model agreed with a large part of the data. Yet, some data on secondary metabolites were not fit well. The posterior parameter distributions obtained for mice were quite narrow (coefficient of variation [CV] of about 10 or 20%), but these CVs might be underestimated because of the incomplete fit of the model. The data fit, for humans, was better than for mice. Yet, some improvement of the model is needed to correctly describe trichloroethanol concentrations over long time periods. Posterior uncertainties about the population means corresponded to 10-20% CV. In terms of human population variability, volumes and flows varied across subject by approximately 20% CV. The variability was somewhat higher for partition coefficients (between 30 and 40%) and much higher for the metabolic parameters (standard deviations representing about a factor of 2). Finally, the analysis points to differences between human males and females in the toxicokinetics of TCE. The significance of these differences in terms of risk remains to be investigated. PMID- 10807560 TI - Statistical analysis of Clewell et al. PBPK model of trichloroethylene kinetics. AB - A physiologically based pharmacokinetic model for trichloroethylene (TCE) in rodents and humans was calibrated with published toxicokinetic data sets. A Bayesian statistical framework was used to combine previous information about the model parameters with the data likelihood, to yield posterior parameter distributions. The use of the hierarchical statistical model yielded estimates of both variability between experimental groups and uncertainty in TCE toxicokinetics. After adjustment of the model by Markov chain Monte Carlo sampling, estimates of variability for the animal or human metabolic parameters ranged from a factor of 1.5-2 (geometric standard deviation [GSD]). Uncertainty was of the same order as variability for animals and higher than variability for humans. The model was used to make posterior predictions for several measures of cancer risk. These predictions were affected by both uncertainties and variability and exhibited GSDs ranging from 2 to 6 in mice and rats and from 2 to 10 for humans. PMID- 10807561 TI - Neurotoxic and pharmacokinetic responses to trichloroethylene as a function of exposure scenario. AB - Strategies are needed for assessing the risks of exposures to airborne toxicants that vary over concentrations and durations. The goal of this project was to describe the relationship between the concentration and duration of exposure to inhaled trichloroethylene (TCE), a representative volatile organic chemical, tissue dose as predicted by a physiologically based pharmacokinetic model, and neurotoxicity. Three measures of neurotoxicity were studied: hearing loss, signal detection behavior, and visual function. The null hypothesis was that exposure scenarios having an equivalent product of concentration and duration would produce equal toxic effects, according to the classic linear form of Haber's Rule ((italic)C(/italic) times t = k), where C represents the concentration, t, the time (duration) of exposure, and k, a constant toxic effect. All experiments used adult male, Long-Evans rats. Acute and repeated exposure to TCE increased hearing thresholds, and acute exposure to TCE impaired signal detection behavior and visual function. Examination of all three measures of neurotoxicity showed that if Haber's Rule were used to predict outcomes across exposure durations, the risk would be overestimated when extrapolating from shorter to longer duration exposures, and underestimated when extrapolating from longer to shorter duration exposures. For the acute effects of TCE on behavior and visual function, the estimated concentration of TCE in blood at the time of testing correlated well with outcomes, whereas cumulative exposure, measured as the area under the blood TCE concentration curve, did not. We conclude that models incorporating dosimetry can account for differing exposure scenarios and will therefore improve risk assessments over models considering only parameters of external exposure. PMID- 10807559 TI - Development of a physiologically based pharmacokinetic model of trichloroethylene and its metabolites for use in risk assessment. AB - A physiologically based pharmacokinetic (PBPK) model was developed that provides a comprehensive description of the kinetics of trichloroethylene (TCE) and its metabolites, trichloroethanol (TCOH), trichloroacetic acid (TCA), and dichloroacetic acid (DCA), in the mouse, rat, and human for both oral and inhalation exposure. The model includes descriptions of the three principal target tissues for cancer identified in animal bioassays: liver, lung, and kidney. Cancer dose metrics provided in the model include the area under the concentration curve (AUC) for TCA and DCA in the plasma, the peak concentration and AUC for chloral in the tracheobronchial region of the lung, and the production of a thioacetylating intermediate from dichlorovinylcysteine in the kidney. Additional dose metrics provided for noncancer risk assessment include the peak concentrations and AUCs for TCE and TCOH in the blood, as well as the total metabolism of TCE divided by the body weight. Sensitivity and uncertainty analyses were performed on the model to evaluate its suitability for use in a pharmacokinetic risk assessment for TCE. Model predictions of TCE, TCA, DCA, and TCOH concentrations in rodents and humans are in good agreement with a variety of experimental data, suggesting that the model should provide a useful basis for evaluating cross-species differences in pharmacokinetics for these chemicals. In the case of the lung and kidney target tissues, however, only limited data are available for establishing cross-species pharmacokinetics. As a result, PBPK model calculations of target tissue dose for lung and kidney should be used with caution. PMID- 10807563 TI - Biologically based dose-response model for liver tumors induced by trichloroethylene. AB - The existing extensive laboratory data on trichloroethylene (TCE) and its two metabolites, dichloroacetic (DCA) and trichloroacetic (TCA), are used to explore the relationship among these three compounds. Under the hypothesis that these compounds induce liver tumors in mice through promotion of preexisting initiated cells, it is demonstrated that DCA alone could be responsible for all the response of carcinomas in liver of B6CF(1) mice. The focus of this paper is on how a plausible biological assumption could impact on low-dose risk estimates, rather than on the risk estimate per se. The findings suggest that low-dose risk estimates to humans would be overestimated unless the different background rates between mice and humans are properly accounted for. PMID- 10807562 TI - Evaluating noncancer effects of trichloroethylene: dosimetry, mode of action, and risk assessment. AB - Alternatives for developing chronic exposure limits for noncancer effects of trichloroethylene (TCE) were evaluated. These alternatives were organized within a framework for dose-response assessment--exposure:dosimetry (pharmacokinetics):mode of action (pharmacodynamics): response. This framework provides a consistent structure within which to make scientific judgments about available information, its interpretation, and use. These judgments occur in the selection of critical studies, internal dose metrics, pharmacokinetic models, approaches for interspecies extrapolation of pharmacodynamics, and uncertainty factors. Potentially limiting end points included developmental eye malformations, liver effects, immunotoxicity, and kidney toxicity from oral exposure and neurological, liver, and kidney effects by inhalation. Each end point was evaluated quantitatively using several methods. Default analyses used the traditional no-observed adverse effect level divided by uncertainty factors and the benchmark dose divided by uncertainty factors methods. Subsequently, mode of-action and pharmacokinetic information were incorporated. Internal dose metrics were estimated using a physiologically based pharmacokinetic (PBPK) model for TCE and its major metabolites. This approach was notably useful with neurological and kidney toxicities. The human PBPK model provided estimates of human exposure doses for the internal dose metrics. Pharmacodynamic data or default assumptions were used for interspecies extrapolation. For liver and neurological effects, humans appear no more sensitive than rodents when internal dose metrics were considered. Therefore, the interspecies uncertainty factor was reduced, illustrating that uncertainty factors are a semiquantitative approach fitting into the organizational framework. Incorporation of pharmacokinetics and pharmacodynamics can result in values that differ significantly from those obtained with the default methods. PMID- 10807564 TI - Dose-response analyses of the carcinogenic effects of trichloroethylene in experimental animals. AB - In lifetime bioassays, trichloroethylene (TCE, CAS No. 79-01-6) causes liver tumors in mice following gavage, liver and lung tumors in mice following inhalation, and kidney tumors in rats following gavage or inhalation. Recently developed pharmacokinetic models provide estimates of internal, target-organ doses of the TCE metabolites thought responsible for these tumor responses. Dose response analyses following recently proposed methods for carcinogen risk assessment from the U.S. Environmental Protection Agency (U.S. EPA) are conducted on the animal tumor data using the pharmacokinetic dosimeters to derive a series of alternative projections of the potential carcinogenic potency of TCE in humans exposed to low environmental concentrations. Although mechanistic considerations suggest action of possibly nonlinear processes, dose-response shapes in the observable range of tumor incidence evince little sign of such patterns. Results depend on which of several alternative pharmacokinetic analyses are used to define target-organ doses. Human potency projections under the U.S. EPA linear method based on mouse liver tumors and internal dosimetry equal or somewhat exceed calculations based on administered dose, and projections based on mouse liver tumors exceed those from mouse lung or rat kidney tumors. Estimates of the carcinogenic potency of the two primary oxidative metabolites of TCE- trichloroacetic acid and dichloroacetic acid, which are mouse liver carcinogens in their own right--are also made, but it is not clear whether the carcinogenic potency of TCE can be quantitatively ascribed to metabolic generation of these metabolites. PMID- 10807565 TI - Exposure assessment of trichloroethylene. AB - This article reviews exposure information available for trichloroethylene (TCE) and assesses the magnitude of human exposure. The primary sources releasing TCE into the environment are metal cleaning and degreasing operations. Releases occur into all media but mostly into the air due to its volatility. It is also moderately soluble in water and can leach from soils into groundwater. TCE has commonly been found in ambient air, surface water, and groundwaters. The 1998 air levels in microg/m(3) across 115 monitors can be summarized as follows: range = 0.01-3.9, mean = 0.88. A California survey of large water utilities in 1984 found a median concentration of 3.0 microg/L. General population exposure to TCE occurs primarily by inhalation and water ingestion. Typical average daily intakes have been estimated as 11-33 microg/day for inhalation and 2-20 microg/day for ingestion. A small portion of the population is expected to have elevated exposures as a result of one or more of these pathways: inhalation exposures to workers involved in degreasing operations, ingestion and inhalation exposures occurring in homes with private wells located near disposal/contamination sites, and inhalation exposures to consumers using TCE products in areas of poor ventilation. More current and more extensive data on TCE levels in indoor air, water, and soil are needed to better characterize the distribution of background exposures in the general population and elevated exposures in special subpopulations. PMID- 10807566 TI - Interacting molecular loops in the mammalian circadian clock. AB - We show that, in the mouse, the core mechanism for the master circadian clock consists of interacting positive and negative transcription and translation feedback loops. Analysis of Clock/Clock mutant mice, homozygous Period2(Brdm1) mutants, and Cryptochrome-deficient mice reveals substantially altered Bmal1 rhythms, consistent with a dominant role of PERIOD2 in the positive regulation of the Bmal1 loop. In vitro analysis of CRYPTOCHROME inhibition of CLOCK: BMAL1 mediated transcription shows that the inhibition is through direct protein:protein interactions, independent of the PERIOD and TIMELESS proteins. PERIOD2 is a positive regulator of the Bmal1 loop, and CRYPTOCHROMES are the negative regulators of the Period and Cryptochrome cycles. PMID- 10807568 TI - Separation of long DNA molecules in a microfabricated entropic trap array. AB - A nanofluidic channel device, consisting of many entropic traps, was designed and fabricated for the separation of long DNA molecules. The channel comprises narrow constrictions and wider regions that cause size-dependent trapping of DNA at the onset of a constriction. This process creates electrophoretic mobility differences, thus enabling efficient separation without the use of a gel matrix or pulsed electric fields. Samples of long DNA molecules (5000 to approximately 160,000 base pairs) were efficiently separated into bands in 15-millimeter-long channels. Multiple-channel devices operating in parallel were demonstrated. The efficiency, compactness, and ease of fabrication of the device suggest the possibility of more practical integrated DNA analysis systems. PMID- 10807567 TI - Earliest Pleistocene hominid cranial remains from Dmanisi, Republic of Georgia: taxonomy, geological setting, and age. AB - Archaeological excavations at the site of Dmanisi in the Republic of Georgia have uncovered two partial early Pleistocene hominid crania. The new fossils consist of a relatively complete cranium and a second relatively complete calvaria from the same site and stratigraphic unit that yielded a hominid mandible in 1991. In contrast with the uncertain taxonomic affinity of the mandible, the new fossils are comparable in size and morphology with Homo ergaster from Koobi Fora, Kenya. Paleontological, archaeological, geochronological, and paleomagnetic data from Dmanisi all indicate an earliest Pleistocene age of about 1.7 million years ago, supporting correlation of the new specimens with the Koobi Fora fossils. The Dmanisi fossils, in contrast with Pleistocene hominids from Western Europe and Eastern Asia, show clear African affinity and may represent the species that first migrated out of Africa. PMID- 10807569 TI - Structure of the hydrated alpha-Al(2)O(3) (0001) surface AB - The physical and chemical properties of the hydrated alpha-Al(2)O(3) (0001) surface are important for understanding the reactivity of natural and synthetic aluminum-containing oxides. The structure of this surface was determined in the presence of water vapor at 300 kelvin by crystal truncation rod diffraction at a third-generation synchrotron x-ray source. The fully hydrated surface is oxygen terminated, with a 53% contracted double Al layer directly below. The structure is an intermediate between alpha-Al(2)O(3) and gamma-Al(OH)(3), a fully hydroxylated form of alumina. A semiordered oxygen layer about 2.3 angstroms above the terminal oxygen layer is interpreted as adsorbed water. The clean alpha Al(2)O(3) (0001) surface, in contrast, is Al terminated and significantly relaxed relative to the bulk structure. These differences explain the different reactivities of the clean and hydroxylated surfaces. PMID- 10807570 TI - Rapid flooding of the sunda shelf: A late-glacial sea-level record AB - The increase in sea level from the last glacial maximum has been derived from a siliciclastic system on the tectonically stable Sunda Shelf in Southeast Asia. The time from 21 to 14 thousand calendar years before the present has been poorly covered in other records. The record generally confirms sea-level reconstructions from coral reefs. The rise of sea level during meltwater pulse 1A was as much as 16 meters within 300 years (14.6 to 14.3 thousand years ago). PMID- 10807571 TI - The nature of pristine noble gases in mantle plumes AB - High-precision noble gas data show that the Hawaiian and Icelandic mantle plume sources contain uniquely primitive neon that is composed of moderately nucleogenic neon-21 and a primordial component indistinguishable from the meteoritic occurrence of solar neon. This suggests that Earth's solar-type rare gas inventory was acquired during accretion from small planetesimals previously irradiated by solar wind from the early sun. However, nonradiogenic argon, krypton, and xenon isotopes derived from the mantle display nonsolar compositions and indicate an atmosphere-like fingerprint that is not due to recent subduction. PMID- 10807572 TI - El Nino-like climate teleconnections in new england during the late pleistocene AB - A glacial varve chronology from New England spanning the 4000-year period from 17,500 to 13,500 calendar years before the present was analyzed for evidence of climate variability during the late Pleistocene. The chronology shows a distinct interannual (3 to 5 years) band of enhanced variability suggestive of El Nino Southern Oscillation (ENSO) teleconnections into North America during the late Pleistocene, when the Laurentide ice sheet was near its maximum extent and climatic boundary conditions were different than those of today. This interannual variability largely disappears by the young end of the 4000-year chronology, with only the highest frequency components (roughly 3-year period) persisting. This record provides evidence of ENSO-like climate variability during near-peak glacial conditions. PMID- 10807573 TI - Reduction of tropical cloudiness by soot AB - Measurements and models show that enhanced aerosol concentrations can augment cloud albedo not only by increasing total droplet cross-sectional area, but also by reducing precipitation and thereby increasing cloud water content and cloud coverage. Aerosol pollution is expected to exert a net cooling influence on the global climate through these conventional mechanisms. Here, we demonstrate an opposite mechanism through which aerosols can reduce cloud cover and thus significantly offset aerosol-induced radiative cooling at the top of the atmosphere on a regional scale. In model simulations, the daytime clearing of trade cumulus is hastened and intensified by solar heating in dark haze (as found over much of the northern Indian Ocean during the northeast monsoon). PMID- 10807574 TI - N-Cadherin, a cell adhesion molecule involved in establishment of embryonic left right asymmetry. AB - Within the bilaterally symmetric vertebrate body plan, many organs develop asymmetrically. Here, it is demonstrated that a cell adhesion molecule, N cadherin, is one of the earliest proteins to be asymmetrically expressed in the chicken embryo and that its activity is required during gastrulation for proper establishment of the left-right axis. Blocking N-cadherin function randomizes heart looping and alters the expression of Snail and Pitx2, later components of the molecular cascade that regulate left-right asymmetry. However, the expression of other components of this cascade (Nodal and Lefty) was unchanged after blocking N-cadherin function, suggesting the existence of parallel pathways in the establishment of left-right morphogenesis. Here, the results suggest that N cadherin-mediated cell adhesion events are required for establishment of left right asymmetry. PMID- 10807575 TI - Influenza B virus in seals. AB - Influenza B virus is a human pathogen whose origin and possible reservoir in nature are not known. An influenza B virus was isolated from a naturally infected harbor seal (Phoca vitulina) and was found to be infectious to seal kidney cells in vitro. Sequence analyses and serology indicated that influenza virus B/Seal/Netherlands/1/99 is closely related to strains that circulated in humans 4 to 5 years earlier. Retrospective analyses of sera collected from 971 seals showed a prevalence of antibodies to influenza B virus in 2% of the animals after 1995 and in none before 1995. This animal reservoir, harboring influenza B viruses that have circulated in the past, may pose a direct threat to humans. PMID- 10807576 TI - Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. AB - A critical function of tumor suppressor p53 is the induction of apoptosis in cells exposed to noxious stresses. We report a previously unidentified pro apoptotic gene, Noxa. Expression of Noxa induction in primary mouse cells exposed to x-ray irradiation was dependent on p53. Noxa encodes a Bcl-2 homology 3 (BH3) only member of the Bcl-2 family of proteins; this member contains the BH3 region but not other BH domains. When ectopically expressed, Noxa underwent BH3 motif dependent localization to mitochondria and interacted with anti-apoptotic Bcl-2 family members, resulting in the activation of caspase-9. We also demonstrate that blocking the endogenous Noxa induction results in the suppression of apoptosis. Noxa may thus represent a mediator of p53-dependent apoptosis. PMID- 10807577 TI - Stable RNA/DNA hybrids in the mammalian genome: inducible intermediates in immunoglobulin class switch recombination. AB - Although it is well established that mammalian class switch recombination is responsible for altering the class of immunoglobulins, the mechanistic details of the process have remained unclear. Here, we show that stable RNA/DNA hybrids form at class switch sequences in the mouse genome upon cytokine-specific stimulation of class switch in primary splenic B cells. The RNA hybridized to the switch DNA is transcribed in the physiological orientation. Mice that constitutively express an Escherichia coli ribonuclease H transgene show a marked reduction in RNA/DNA hybrid formation, an impaired ability to generate serum immunoglobulin G antibodies, and significant inhibition of class switch recombination in their splenic B cells. These data provide evidence that stable RNA/DNA hybrids exist in the mammalian nuclear genome, can serve as intermediates for physiologic processes, and are mechanistically important for efficient class switching in vivo. PMID- 10807578 TI - A high-affinity iron permease essential for Candida albicans virulence. AB - Microbial pathogens must compete with the iron-withholding defense systems of their host to acquire this essential nutrient. Here, two high-affinity iron permease genes, CaFTR1 and CaFTR2, were isolated. CaFTR1 expression was induced under iron-limited conditions and repressed when iron supply was sufficient, whereas the expression of CaFTR2 was regulated in a reversed manner. Mutants lacking CaFTR1 but not CaFTR2 exhibited a severe growth defect in iron-deficient medium and were unable to establish systemic infection in mice. Thus, CaFTR1 mediated iron-uptake mechanism constitutes a virulence factor of Candida albicans and may be a target for the development of anti-Candida therapies. PMID- 10807580 TI - Pushing, pulling, dragging, and vibrating renal epithelia by using atomic force microscopy. AB - Renal physiologists focus on events that take place on and around the surfaces of cells. Various techniques have been developed that follow transport functions at the molecular level, but until recently none of these techniques has been capable of making the behavior of molecular structures visible under physiological conditions. This apparent gap may be filled in the future by the application of atomic force microscopy. This technique produces an image not by optical means, but by "feeling" its way across a surface. Atomic force microscopy can, however, be modified in a number of ways, which means that besides producing a high resolution image, it is possible to obtain several types of data on the interactions between the ultrastructural components of cell membranes (such as proteins) and other biologically active molecules (such as ATP). In this review we describe the recent use of the atomic force microscope in renal physiology, ranging from experiments in intact cells to those in isolated renal transport protein molecules, include examples of these extended applications of the technique, and point to uses that the microscope has recently found in other areas of biology that should prove fruitful in renal physiology in the near future. PMID- 10807581 TI - Localization of protein inhibitor of neuronal nitric oxide synthase in rat kidney. AB - We have recently demonstrated that in rats with 5/6 nephrectomy (5/6 Nx), renal cortical and inner medullary neuronal NOS (nNOS) expression is downregulated, associated with decreased urinary excretion of nitric oxide (NO) products. Recently, a novel 89-amino acid protein [protein inhibitor of nNOS (PIN)] was isolated from rat brain and shown to inhibit nNOS activity. The present studies localized PIN in the rat kidney and determined the effect of 5/6 Nx on PIN expression. By RT-PCR, PIN mRNA was detected in the kidney cortex and inner medulla. Immunohistochemistry revealed staining for PIN in glomerular and vasa rectae endothelial cells. PIN was also localized to the apical membranes of inner medullary collecting duct (IMCD) cells. Two weeks after 5/6 Nx, inner medullary PIN expression was significantly upregulated (sham, 0.18+/-0.07 vs. 5/6 Nx, 0.58+/-0.13 arbitrary units; n = 6, P<0.02), as determined by Western blotting. In summary, our data show that PIN, a specific inhibitor of nNOS activity, is expressed in the IMCD, a site of high nNOS expression in the kidney. PIN expression is upregulated in the inner medulla of 5/6 Nx rats. Inhibition of nNOS activity by PIN may have important implications for the regulation of NO synthesis in the IMCD of normal and remnant kidneys. PMID- 10807582 TI - Angiotensinogen and AT(1) antisense inhibition of osteopontin translation in rat proximal tubular cells. AB - Antisense oligonucleotide inhibition of angiotensinogen and ANG II type 1 receptor (AT(1)) mRNA translation in rat proximal tubules (PT) was examined to provide direct evidence for a role of the renin-angiotensin system (RAS) in upregulated osteopontin expression observed following mechanical cell stretch. Male Sprague-Dawley rats underwent unilateral ureteral obstruction (UUO) under Brevital anesthesia. In situ hybridization and Western blot analysis demonstrated angiotensinogen mRNA and angiotensin converting enzyme (ACE) protein localized to PTs and upregulated in obstructed kidneys, respectively, confirming an increased expression of renal RAS in vivo. In vitro studies were performed to provide mechanistic insight into ANG II-dependent osteopontin expression following mechanical cell stretch, which putatively mimics the increased PT luminal pressure post-UUO. A cationic transfection method was used to introduce either angiotensinogen or AT(1) antisense oligonucleotide into cultured rat PT cells prior to 1 h of cyclic mechanical cell stretch. Northern blot analysis revealed that PT cells subjected to cyclic mechanical stretch with/without prior transfection with a sense oligonucleotide exhibited increased osteopontin mRNA expression compared with unstretched cells. Blockade of either angiotensinogen or AT(1) mRNA translation by antisense oligonucleotide inhibition prior to cell stretch was found to significantly decrease osteopontin mRNA levels 2.4-fold (P<0.004) and 1.6-fold (P<0.001), respectively, compared with values observed in control unstretched cells. This study provides evidence that stretch-induced upregulation of osteopontin mRNA expression is mediated, in part, via production of ANG II. These results lend insight into upregulation of osteopontin via a local PT RAS leading to macrophage infiltration in the tubulointerstitium in experimental hydronephrosis. PMID- 10807584 TI - Heme oxygenase-1 gene ablation or expression modulates cisplatin-induced renal tubular apoptosis. AB - Heme oxygenase-1 (HO-1) is a 32-kDa microsomal enzyme that catalyzes the conversion of heme to biliverdin, releasing iron and carbon monoxide. Induction of HO-1 occurs as a protective response in cells/tissues exposed to a wide variety of oxidant stimuli. The chemotherapeutic effects of cis diamminedichloroplatinum(II) (cisplatin), a commonly used anticancer drug, are limited by significant nephrotoxicity, which is characterized by varying degrees of renal tubular apoptosis and necrosis. The purpose of this study was to evaluate the functional significance of HO-1 expression in cisplatin-induced renal injury. Our studies demonstrate that transgenic mice deficient in HO-1 (-/ ), develop more severe renal failure and have significantly greater renal injury compared with wild-type (+/+) mice treated with cisplatin. In vitro studies in human renal proximal tubule cells demonstrate that hemin, an inducer of HO-1, significantly attenuated cisplatin-induced apoptosis and necrosis, whereas inhibition of HO-1 enzyme activity reversed the cytoprotective effect. Overexpression of HO-1 resulted in a significant reduction in cisplatin-induced cytotoxicity. These studies provide a basis for future studies using targeted gene expression of HO-1 as a therapeutic and preventive modality in high-risk settings of acute renal failure. PMID- 10807585 TI - Similarities and differences in the subcellular localization of hamartin and tuberin in the kidney. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder characterized by hamartomas in multiple organs, notably the brain and kidneys. The disease is caused by mutations in TSC1 or TSC2 genes, coding hamartin and tuberin, respectively. Immunofluorescence analysis of tuberin and hamartin performed here demonstrates that both proteins are specifically expressed in the distal urinary tubule, comprising the distal tubules, connecting segment, and collecting ducts. Hamartin, distinct from tuberin, is expressed in the thick ascending limbs of Henle and in juxtaglomerular cells, where it colocalizes with renin. In positive epithelial cells, tuberin localizes to the cytoplasm as well as the apical membrane. Hamartin, however, preferentially localizes to the apical membrane. The two proteins colocalize at the apical membrane of type A intercalated cells and connecting tubule cells, whereas in type B intercalated cells they reveal a variable pattern of expression. The cell-specific expression of tuberin and hamartin described here will provide critical insight into the cell types that give rise to kidney lesions, and the tumor suppressor role of these proteins in TSC. PMID- 10807583 TI - Tetanus toxin-mediated cleavage of cellubrevin inhibits proton secretion in the male reproductive tract. AB - Our laboratory has previously shown that the vacuolar H(+)-ATPase, located in a subpopulation of specialized cells establishes a luminal acidic environment in the epididymis and proximal part of the vas deferens (Breton S, Smith PJS, Lui B, and Brown D. Nat Med 2: 470-472, 1996). Low luminal pH is critical for sperm maturation and maintenance of sperm in a quiescent state during storage in these organs. In the present study we examined the regulation of proton secretion in the epididymis and vas deferens. In vivo microtubule disruption by colchicine induced an almost complete loss of H(+)-ATPase apical polarity. Endocytotic vesicles, visualized by Texas red-dextran internalization, contain H(+)-ATPase, indicating active endocytosis of the pump. Cellubrevin, an analog of the vesicle soluble N-ethyl malemide-sensitive factor attachment protein (SNAP) receptor (v SNARE) synaptobrevin, is highly enriched in H(+)-ATPase-rich cells of the epididymis and vas deferens, and tetanus toxin treatment markedly inhibited bafilomycin-sensitive proton secretion by 64.3+/-9.0% in the proximal vas deferens. Western blotting showed effective cleavage of cellubrevin by tetanus toxin in intact vas deferens, demonstrating that the toxin gained access to cellubrevin. These results suggest that H(+)-ATPase is actively endocytosed and exocytosed in proton-secreting cells of the epididymis and vas deferens and that net proton secretion requires the participation of the v-SNARE cellubrevin. PMID- 10807588 TI - Roles of NO and oxygen radicals in tubuloglomerular feedback in SHR. AB - The spontaneously hypertensive rat (SHR) has enhanced tubuloglomerular feedback (TGF) responses and diminished buffering by juxtaglomerular apparatus (JGA) derived nitric oxide (NO) despite enhanced expression of NO synthase (NOS) isoforms in the JGA. We tested the hypothesis that the enhanced TGF response is due to inactivation of NO by oxygen radicals (O(-)(2)). SHR had significantly (P<0.05) greater expression of the peroxynitrate reaction product, nitrotyrosine, in renal cortex. A membrane-permeant, metal-independent superoxide dismutase mimetic, tempol, was used to test the functional role of O(-)(2). Maximum TGF responses, assessed from changes in proximal stop-flow pressure (P(SF)) during orthograde loop of Henle (LH) perfusion of artificial tubular fluid (ATF), were enhanced in SHR [Wistar-Kyoto rat (WKY) 8.8+/-0.4 (n = 30 nephrons) vs. SHR 10.8+/-0.4 mm Hg (n = 39 nephrons), P<0.001]. TGF responses of SHR were unresponsive to microperfusion of 7-nitroindazole (7-NI, 10(-4) M), which is an inhibitor of neuronal NOS (nNOS) [WKY 8.3+/-0.3 to 10.8+/-0.4 (n = 8, P<0.001) vs. SHR 10.0+/-0.7 to 10.5+/-0.8 mm Hg (n = 8; not significant)]. Microperfusion of tempol (10(-4) M) into the efferent arteriole (EA) supplying the peritubular capillaries (PTC) blunted TGF. The response to tempol was significantly (P< 0.05) greater in SHR [DeltaTGF in WKY 19+/-6% (n = 10) vs. SHR 32+/-3% (n = 10)]. Microperfusion of the NO donor compound S-nitroso-N-acetyl-penicillamine (SNAP, 10(-7)-10(-4) M) via the LH blunted TGF, but the sensitivity of the response was impaired significantly (P<0.05) in SHR nephrons. PTC perfusion of tempol (10(-4) M) normalized the response to loop perfusion of both SNAP and 7-NI in SHR nephron to levels in WKY (during tempol, DeltaP(SF) with 7-NI in WKY 8.9+/-0.6 to 11.4+/ 0.8; n = 12 vs. SHR 9.5+/-0.5 to 12.5+/-0.4 mm Hg; n = 16). In conclusion, TGF responses are enhanced in SHR, in part due to a diminished role for NO from nNOS in blunting TGF due to enhanced O(-)(2) formation. O(-)(2) in the JGA enhances TGF responses by inactivation of locally generated NO. PMID- 10807587 TI - Mouse proximal tubular cell-cell adhesion inhibits apoptosis by a cadherin dependent mechanism. AB - Adhesion of epithelial cells to matrix is known to inhibit apoptosis. However, the role of cell-cell adhesion in mediating cell survival remains uncertain. Primary cultures of mouse proximal tubular (MPT) cells were used to examine the role of cell-cell adhesion in promoting survival. When MPT cells were deprived of both cell-matrix and cell-cell adhesion, they died by apoptosis. However, when incubated in agarose-coated culture dishes (to prevent cell-matrix adhesion) and at high cell density (to allow cell-cell interactions), MPT cells adhered to one another and remained viable. Expression of E-cadherin among suspended, aggregating cells increased with time. A His-Ala-Val (HAV)-containing peptide that inhibits homophilic E-cadherin binding prevented cell-cell aggregation and promoted apoptosis of MPT cells in suspension. By contrast, inhibition of potential beta(1)-integrin-mediated interactions between cells in suspension did not prevent either aggregation or survival of suspended cells. Aggregation of cells in suspension activated phosphatidylinositol 3-kinase (PI3K), an event that was markedly reduced by the presence of the HAV peptide. LY-294002, an inhibitor of PI3K, also inhibited survival of suspended cells. In summary, we provide novel evidence that MPT cells, when deprived of normal cell-matrix interactions, can adhere to one another in a cadherin-dependent fashion and remain viable. Survival of aggregated cells depends on activation of PI3K. PMID- 10807586 TI - C5b-9 membrane attack complex mediates endothelial cell apoptosis in experimental glomerulonephritis. AB - We studied the role of the C5b-9 membrane attack complex in two models of inflammatory glomerulonephritis (GN) initiated by acute glomerular endothelial injury in Piebold-viral-Glaxo (PVG) complement-sufficient rats (C+), C6-deficient rats (C6-), and rats systematically depleted of complement with cobra venom factor (CVF). GN was induced by performing a left nephrectomy and selectively perfusing the right kidney with either 1) the lectin concanavalin A (Con A) followed by complement-fixing anti-Con A (Con A GN) or 2) purified complement fixing goat anti-rat glomerular endothelial cell (GEN) antibody [immune-mediated thrombotic microangiopathy (ITM)]. Comparable levels of GEN apoptosis were detected in C+ animals in both models. CVF administration reduced GEN apoptosis by 10- to 12-fold. GEN apoptosis was C5b-9 dependent because PVG C6- rats were protected from GEN loss. Furthermore, functional inhibition of the cell surface complement regulatory protein CD59 by renal perfusion with anti-CD59 antibody in ITM resulted in a 3.5-fold increase in GEN apoptosis. Last, in Con A GN, abrogation of GEN apoptosis preserved endothelial integrity and renal function. This study demonstrates the specific role of C5b-9 in the induction of GEN apoptosis in experimental inflammatory GN, a finding with implications for diseases associated with the presence of antiendothelial cell antibodies. PMID- 10807589 TI - Water transport by the renal Na(+)-dicarboxylate cotransporter. AB - This study investigated the ability of the renal Na(+)-dicarboxylate cotransporter, NaDC-1, to transport water. Rabbit NaDC-1 was expressed in Xenopus laevis oocytes, cotransporter activity was measured as the inward current generated by substrate (citrate or succinate), and water transport was monitored by the changes in oocyte volume. In the absence of substrates, oocytes expressing NaDC-1 showed an increase in osmotic water permeability, which was directly correlated with the expression level of NaDC-1. When NaDC-1 was transporting substrates, there was a concomitant increase in oocyte volume. This solute coupled influx of water took place in the absence of, and even against, osmotic gradients. There was a strict stoichiometric relationship between Na(+), substrate, and water transport of 3 Na(+), 1 dicarboxylate, and 176 water molecules/transport cycle. These results indicate that the renal Na(+) dicarboxylate cotransporter mediates water transport and, under physiological conditions, may contribute to fluid reabsorption across the proximal tubule. PMID- 10807590 TI - Novel biochemical and functional insights into nuclear Ca(2+) transport through IP(3)Rs and RyRs in osteoblasts. AB - We report the first biochemical and functional characterization of inositol trisphosphate receptors (IP(3)Rs) and ryanodine receptors (RyRs) in the nuclear membrane of bone-forming (MC3T3-E1) osteoblasts. Intact nuclei fluoresced intensely with anti-RyR (Ab(34)) and anti-IP(3)R (Ab(40)) antisera in a typically peripheral nuclear membrane pattern. Isolated nuclear membranes were next subjected to SDS-PAGE and blotted with isoform-specific anti-receptor antisera, notably Ab(40), anti-RyR-1, anti-RyR-2 (Ab(129)), and anti-RyR-3 (Ab(180)). Only anti-RyR-1 and Ab(40) showed bands corresponding, respectively, to full-length RyR-1 ( approximately 500 kDa) and IP(3)R-1 (approximately 250 kDa). Band intensity was reduced by just approximately 20% after brief tryptic proteolysis of intact nuclei; this confirmed that isolated nuclear membranes were mostly free of endoplasmic reticular contaminants. Finally, the nucleoplasmic Ca(2+) concentration ([Ca(2+)](np)) was measured in single nuclei by using fura-dextran. The nuclear envelope was initially loaded with Ca(2+) via Ca(2+)-ATPase activation (1 mM ATP and approximately 100 nM Ca(2+)). Adequate Ca(2+) loading was next confirmed by imaging the nuclear envelope (and nucleoplasm). Exposure of Ca(2+)-loaded nuclei to IP(3) or cADP ribose resulted in a rapid and sustained [Ca(2+)](np) elevation. Taken together, the results provide complementary evidence for nucleoplasmic Ca(2+) influx in osteoblasts through nuclear membrane resident IP(3)Rs and RyRs. Our findings may conceivably explain the direct regulation of osteoblastic gene expression by hormones that use the IP(3)-Ca(2+) pathway. PMID- 10807591 TI - Luminal and contraluminal action of 1-34 and 3-34 PTH peptides on renal type IIa Na-P(i) cotransporter. AB - Parathyroid hormone (PTH) inhibits proximal tubular reabsorption of P(i) by retrieval of type IIa Na-P(i) cotransporters (NaPi-IIa) from the brush-border membrane (BBM). We analyzed by immunohistochemistry whether PTH analogs, signaling through either protein kinase A (PKA) and C (PKC; 1-34 PTH) or only PKC (3-34 PTH), elicit in rat kidney in vivo or in the perfused murine proximal tubule in vitro a retrieval of NaPi-IIa and whether pharmacological agonists or inhibitors of these kinases are able to either mimic or interfere with these PTH effects. Treatment with either 1-34 or 3-34 PTH downregulated NaPi-IIa in rat kidney. In isolated murine proximal tubules 1-34 PTH was effective when added to either the apical or basolateral perfusate, whereas 3-34 PTH acted only via the luminal perfusate. These effects were mimicked by an activation of PKA with 8 bromoadenosine 3',5'-cyclic monophosphate or PKC with 1, 2-dioctanoylglycerol. The luminal action of both PTH peptides was blocked by inhibition of the PKC pathway (calphostin C), whereas the basolateral effect of 1-34 PTH was completely abolished by inhibiting both pathways (H-89 and calphostin C). These results suggest that 1) NaPi-IIa can be internalized by cAMP-dependent and -independent signaling mechanisms; 2) functional PTH receptors are located in both membrane domains; and 3) apical PTH receptors may preferentially initiate the effect through a PKC-dependent mechanism. PMID- 10807592 TI - Chlorpropamide upregulates antidiuretic hormone receptors and unmasks constitutive receptor signaling. AB - The mechanism by which chlorpropamide (CP) treatment promotes antidiuresis is unknown. CP competitively inhibited antidiuretic hormone (ADH) binding and adenylyl cyclase (AC) stimulation (inhibition constants K(i) and K'(i) of 2.8 mM and 250 microM, respectively) in the LLC-PK(1) cell line. CP (333 microM) increased the apparent K(a) of ADH for AC activation (0.31 vs. 0.08 nM) without affecting a maximal response, suggesting competitive antagonism. Because CP lowers "basal" AC activity and the AC activation-ADH receptor occupancy relationship (A-O plots), it is an ADH inverse agonist. Twenty-four-hour CP exposure (100 microM) upregulated the ADH receptors without affecting affinity. This lowered K(a) and increased basal AC activity and maximal response (1. 86 vs. 1.35 and 14.9 vs. 10.6 fmol cAMP. min(-1). 10(3) cells(-1), n = 6, P<0.05). NaCl, which potentiates ADH stimulation, also increased basal AC activity. This, together with the CP-ADH inverse agonism and increased basal AC activity at higher receptor density, unmasks constitutive receptor signaling. The CP-ADH inverse agonism explains receptor upregulation and predicts the need for residual ADH with functional isoreceptors for CP-mediated antidiuresis. This could be why CP ameliorates partial central diabetes insipidus but not nephrogenic diabetes insipidus. PMID- 10807593 TI - V(1) receptors in luminal action of vasopressin on distal K(+) secretion. AB - Luminal perfusion with collected proximal fluid increases distal K(+) secretion compared with artificial solutions. Arginine vasopressin (AVP), present in luminal fluid, might be responsible for this observation. K(+) secretion rate (J(K)) was measured by K(+)-sensitive microelectrodes during paired luminal stationary microperfusion with control and AVP-containing 0.5 mM K(+) solutions. J(K) was 1.34 +/- 0.35 (n = 24 tubules) nmol x cm(-2) x s(-1) during perfusion with 10(-9) M AVP, against 0.90+/-0.12 nmol x cm(-2) x s(-1) (n = 21) in control (P<0.02). With 10(-9) M AVP+10(-6) M beta-mercapto-beta-beta-cyclopenta methylenepropionyl(1), O-Me-Tyr(2)-Arg(8) vasopressin (MCMV), a specific peptide V(1)-receptor antagonist, J(K) was 0.36+/-0.067 against 0.77+/-0.10 (control; n = 9) nmol x cm(-2) x s(-1) (P<0.01). With 10(-6) M MCMV alone, J(K) was 0.37+/-0.04 against a control of 0.62+/-0.06 (n = 19) nmol. cm(-2). s(-1) (P<0.01). A peptide V(2) antagonist had no such effect. In Brattleboro rats, which do not produce endogenous AVP, MCMV had no effect when given alone, although AVP still stimulated J(K). In conclusion, luminal AVP stimulates distal J(K) significantly. The V(1) antagonist MCMV inhibits the effect of AVP but also reduces J(K) when given alone. This suggests that AVP acts luminally via V(1) receptors but also that there appears to be a background effect of endogenous AVP blocked by the antagonist. PMID- 10807594 TI - Glomerular hemodynamics in severe obesity. AB - Differential solute clearances were used to characterize glomerular function in 12 nondiabetic subjects with severe obesity (body mass index >38). Nine healthy subjects served as the control group. In the obese group, glomerular filtration rate (GFR) and renal plasma flow (RPF) exceeded the control value by 51 and 31%, respectively. Consequently, filtration fraction increased. The augmented RPF suggested a state of renal vasodilatation involving, mainly or solely, the afferent arteriole. Albumin excretion rate and fractional albumin clearance increased by 89 and 78%, respectively. Oral glucose tolerance tests were suggestive of insulin resistance. Insulin resistance was positively correlated with GFR (r = 0.88, P<0.001) and RPF (r = 0.72, P <0.001). Mean arterial pressure was higher than in the control group. Fractional clearances of dextrans of broad size distribution tended to be lowered. The determinants of the GFR were estimated qualitatively by using a theoretical model of dextran transport through a heteroporous membrane. This analysis suggests that the high GFR in very obese subjects may be the result of an increase in transcapillary hydraulic pressure difference (DeltaP). An abnormal transmission of increased arterial pressure to the glomerular capillaries through a dilated afferent arteriole could account for the augmentation in DeltaP. PMID- 10807595 TI - Arachidonic acid metabolic pathways regulating activity of renal Na(+)-K(+) ATPase are age dependent. AB - Locally formed arachidonic acid (AA) metabolites are important as modulators of many aspects of renal tubular function, including regulation of the activity of tubular Na(+)-K(+)-ATPase. Here we examined the ontogeny of the AA metabolic pathways regulating proximal convoluted tubular (PCT) Na(+)-K(+)-ATPase activity in infant and adult rats. Eicosatetraynoic acid, an inhibitor of all AA metabolizing pathways, abolished this effect. AA inhibition of PCT Na(+)-K(+) ATPase was blocked by the 12-lipoxygenase inhibitor baicalein in infant but not in adult rats and by the specific cytochrome P-450 fatty acid omega-hydroxylase inhibitor 17-octadecynoic acid in adult but not in infant rats. The lipoxygenase metabolite 12(S)-hydroxyeicosatetraenoic acid (HETE) and the cytochrome P-450 metabolite 20-HETE both inhibited PCT Na(+)-K(+)-ATPase in a protein kinase C dependent manner, but the effect was significantly more pronounced in infant PCT. Lipoxygenase mRNA was only detected in infant cortex. Expression of renal isoforms of cytochrome P-450 mRNA was more prominent in adult cortex. In summary, the AA metabolic pathways that modulated the activity of rat renal proximal tubular Na(+)-K(+)-ATPase are age dependent. PMID- 10807596 TI - Stimulation of TGF-beta type II receptor by high glucose in mouse mesangial cells and in diabetic kidney. AB - Transforming growth factor-beta (TGF-beta) is important in the pathogenesis of diabetic nephropathy, but little is known about the regulation of the ligand binding TGF-beta type II signaling receptor (TbetaIIR). There were significant increases in TbetaIIR protein and mRNA levels in kidney cortex after 1-6 wk of streptozotocin-induced diabetes. Mouse mesangial cells cultured in high glucose demonstrated significantly increased TbetaIIR protein and mRNA levels compared with normal glucose. This effect was independent of stimulation of TGF-beta bioactivity by high glucose. Consistent with transcriptional activation by high glucose, the half-life ( approximately 4 h) of TbetaIIR mRNA was not affected by glucose concentration. Moreover, mouse mesangial cells transiently transfected with reporter constructs containing the first 47- or 274-bp promoter fragments of TbetaIIR demonstrated significantly increased reporter activity in high glucose. Cells grown in high glucose demonstrated increased responsiveness to a relatively small dose of exogenous TGF-beta(1) (0.5 ng/ml): [(3)H]proline incorporation and alpha(1)(IV) collagen mRNA were significantly greater in cells cultured in high than in normal glucose. Hence, the expression of TbetaIIR is increased in the diabetic kidney and in mesangial cells cultured in high glucose, primarily because of stimulation of gene transcription. TbetaIIR upregulation by high ambient glucose may contribute to the increased sensitivity of mesangial cells to the profibrogenic action of TGF-beta(1). PMID- 10807597 TI - Onset of glomerular hypertension with aging precedes injury in the spontaneously hypertensive rat. AB - The changes in renal hemodynamics that develop with aging in spontaneously hypertensive rats (SHR) were examined. Micropuncture studies revealed that glomerular capillary pressure was elevated in SHR at 9 mo of age compared with 3 mo-old SHR and 9-mo-old normotensive Wistar-Kyoto rats. Glomerular hypertension developed because of a small increase in systemic blood pressure and a decline in preglomerular vascular resistance, allowing transmission of elevated systemic pressure to the glomerular capillaries. The hemodynamic alterations were not a compensatory response to injury, inasmuch as vascular and glomerular morphology were normal in 9-mo-old SHR. To determine the mechanism of these changes, the activity of several vasoactive systems was examined. Similar changes in renal hemodynamics were observed in young and old SHR after blockade of nitric oxide production and after intravenous administration of endothelin. However, ANG II produced a proportionally greater reduction in glomerular filtration rate than renal blood flow in older SHR. These data suggest that reduced endogenous activity of the renin-angiotensin system leads to glomerular hypertension in aging SHR. Late development of glomerular hypertension may contribute to the subsequent appearance of glomerular sclerosis and progressive renal failure in these rats. PMID- 10807598 TI - Selective degradation of E-cadherin and dissolution of E-cadherin-catenin complexes in epithelial ischemia. AB - Ischemic epithelial cells are characterized by disruption of intercellular junctions and loss of apical-basolateral protein polarity, which are normally dependent on the integrity of the adherens junction (AJ). Biochemical analysis of both whole ischemic kidneys and ATP-depleted Madin-Darby canine kidney (MDCK) cells demonstrated a striking loss of E-cadherin (the transmembrane protein of the AJ) with the appearance and accumulation of an approximately 80-kDa fragment reactive with anti-E-cadherin antibodies on Western blots of ATP-depleted MDCK cells. This apparent ischemia-induced degradation of E-cadherin was not blocked by either inhibitors of the major proteolytic pathways (i.e., proteasome, lysosome, or calpain), or by chelation of intracellular calcium, suggesting the involvement of a protease capable of functioning at low ATP and low calcium levels. Immunocytochemistry revealed the movement of several proteins normally comprising the AJ, including E-cadherin and beta-catenin, away from lateral portions of the plasma membrane to intracellular sites. Moreover, rate-zonal centrifugation and immunoprecipitation with anti-E-cadherin and anti-beta-catenin antibodies indicated that ATP depletion disrupted normal E-cadherin-catenin interactions, resulting in the dissociation of alpha- and gamma-catenin from E cadherin and beta-catenin-containing complexes. Because the generation and maintenance of polarized epithelial cells are dependent upon E-cadherin-mediated cell-cell adhesion and normal AJ function, we propose that the rapid degradation of E-cadherin and dissolution of the AJ is a key step in the development of the ischemic epithelial cell phenotype. Furthermore, we hypothesize that the reassembly of the AJ after ischemia/ATP depletion may require a novel bioassembly mechanism involving recombination of newly synthesized and sorted E-cadherin with preexisting pools of catenins that have (temporally) redistributed intracellularly. PMID- 10807599 TI - Babies and consent: yet another NHS scandal. But it should lead to improvements in research governance within the NHS. PMID- 10807600 TI - Health inequalities in women and men. PMID- 10807601 TI - Depression in Parkinson's disease. PMID- 10807602 TI - The health hazards of mobile phones. PMID- 10807604 TI - Hospital criticised for not obtaining proper consent PMID- 10807603 TI - Guidelines for managing acute bacterial meningitis. PMID- 10807605 TI - Mbeki appoints team to look at cause of AIDS. PMID- 10807606 TI - Israeli surgeon accused of killing two patients. PMID- 10807608 TI - In brief PMID- 10807607 TI - Israel's doctors take industrial action. PMID- 10807609 TI - NICE issues guidance on using coronary artery stents. PMID- 10807611 TI - Patients will be paramount in assessments, says CHI. PMID- 10807610 TI - NICE approves Taxol for ovarian cancer. PMID- 10807612 TI - Cyanide spill could have long term impact. PMID- 10807613 TI - EU to provide incentives for "orphan" drugs. PMID- 10807614 TI - Medical bills account for 40% of bankruptcies. PMID- 10807615 TI - Canadians grow dissatisfied with their healthcare system. PMID- 10807616 TI - WMA to produce guidelines on health databases. PMID- 10807617 TI - Cell preselection is key to gene therapy success PMID- 10807618 TI - Nine hospital patients may have been exposed to CJD. PMID- 10807619 TI - Randomised, double blind, placebo controlled study of fluticasone propionate in patients with moderate to severe chronic obstructive pulmonary disease: the ISOLDE trial. AB - OBJECTIVES: To determine the effect of long term inhaled corticosteroids on lung function, exacerbations, and health status in patients with moderate to severe chronic obstructive pulmonary disease. DESIGN: Double blind, placebo controlled study. SETTING: Eighteen UK hospitals. PARTICIPANTS: 751 men and women aged between 40 and 75 years with mean forced expiratory volume in one second (FEV(1)) 50% of predicted normal. INTERVENTIONS: Inhaled fluticasone propionate 500 microgram twice daily from a metered dose inhaler or identical placebo. MAIN OUTCOME MEASURES: Efficacy measures: rate of decline in FEV(1) after the bronchodilator and in health status, frequency of exacerbations, respiratory withdrawals. Safety measures: morning serum cortisol concentration, incidence of adverse events. RESULTS: There was no significant difference in the annual rate of decline in FEV(1 )(P=0.16). Mean FEV(1) after bronchodilator remained significantly higher throughout the study with fluticasone propionate compared with placebo (P<0.001). Median exacerbation rate was reduced by 25% from 1.32 a year on placebo to 0.99 a year on with fluticasone propionate (P=0.026). Health status deteriorated by 3.2 units a year on placebo and 2.0 units a year on fluticasone propionate (P=0.0043). Withdrawals because of respiratory disease not related to malignancy were higher in the placebo group (25% v 19%, P=0.034). CONCLUSIONS: Fluticasone propionate 500 microgram twice daily did not affect the rate of decline in FEV(1) but did produce a small increase in FEV(1). Patients on fluticasone propionate had fewer exacerbations and a slower decline in health status. These improvements in clinical outcomes support the use of this treatment in patients with moderate to severe chronic obstructive pulmonary disease. PMID- 10807620 TI - Comparing health inequality in men and women: prospective study of mortality 1986 96. AB - OBJECTIVES: To study prospectively the differences in health inequality in men and women from 1986-96 using the Office for National Statistics' longitudinal study and new socioeconomic classification. To assess the relative importance of social class (based on employment characteristics) and social position according to the general social advantage of the household to mortality risk in men and women. DESIGN: Prospective study. SETTING: England and Wales. SUBJECTS: Men and women of working age at the time of the 1981 census, with a recorded occupation. MAIN OUTCOME MEASURES: Mortality. RESULTS: In men, social class based on employment relations, measured according to the Office for National Statistics' socioeconomic classification, was the most important influence on mortality. In women, social class based on individual employment relations and conditions showed only a weak gradient. Large differences in risk of mortality in women were found, however, when social position was measured according to the general social advantage in the household. CONCLUSIONS: Comparisons of the extent of health inequality in men and women are affected by the measures of social inequality used. For women, even those in paid work, classifications based on characteristics of the employment situation may give a considerable underestimate. The Office for National Statistics' new measure of socioeconomic position is useful for assessing health inequality in men, but in women a more important predictor of mortality is inequality in general social advantage of the household. PMID- 10807621 TI - The right place at the right time PMID- 10807622 TI - Size and quality of randomised controlled trials in head injury: review of published studies. AB - OBJECTIVE: To assess whether trials in head injury are large enough to avoid moderate random errors and designed to avoid moderate biases. DESIGN: All randomised controlled trials on the treatment and rehabilitation of patients with head injury published before December 1998 were surveyed. Trials were identified from electronic databases, by hand searching journals and conference proceedings, and by contacting researchers. Data were extracted on the number of participants, quality of concealment of allocation, use of blinding, loss to follow up, and types of participants, interventions, and outcome measures. RESULTS: 279 reports were identified, containing information on 208 separate trials. The average number of participants per trial was 82, with no evidence of increasing size over time. The total number of randomised participants in the 203 trials in which size was reported was 16 613. No trials were large enough to detect reliably a 5% absolute reduction in the risk of death or disability, and only 4% were large enough to detect an absolute reduction of 10%. Concealment of allocation was adequate in 22 and inadequate or unclear in 25 of the 47 (23%) in which it was reported. Of 126 trials assessing disability, 111 reported the number of patients followed up, and average loss to follow up was 19%. Of trials measuring disability, 26 (21%) reported that outcome assessors were blinded. CONCLUSIONS: Randomised trials in head injury are too small and poorly designed to detect or refute reliably moderate but clinically important benefits or hazards of treatment. Limited funding for injury research and unfamiliarity with issues of consent may have been important obstacles. PMID- 10807623 TI - Suicidal ideation among outpatients at general neurology clinics: prospective study. PMID- 10807624 TI - Home collection of urine for culture from infants by three methods: survey of parents' preferences and bacterial contamination rates. PMID- 10807625 TI - Email submissions from outside the united kingdom PMID- 10807627 TI - On ageing: selfishness PMID- 10807626 TI - Controversy in primary care: Should asymptomatic haemochromatosis be treated? PMID- 10807628 TI - The pleural cavity. PMID- 10807629 TI - Lesson of the week: intracerebral haemorrhage in young adults: the emerging importance of drug misuse. PMID- 10807630 TI - Medicine: art or science? PMID- 10807631 TI - Matthew and adam PMID- 10807632 TI - ABC of arterial and vascular disease: vasculitis. PMID- 10807633 TI - A promising start, but fundamental reform is needed. PMID- 10807634 TI - Thyroid function tests-time for a reassessment. PMID- 10807635 TI - Economic note: cost of illness studies. PMID- 10807636 TI - High road to independence PMID- 10807637 TI - Obituaries PMID- 10807638 TI - The scientification of love PMID- 10807639 TI - Men's health PMID- 10807640 TI - The nurture assumption: why children turn out the way they Do PMID- 10807641 TI - American psycho PMID- 10807642 TI - A cry of pain and anger PMID- 10807643 TI - Head injury PMID- 10807644 TI - The two tier syndrome behind waiting lists PMID- 10807646 TI - How to run the NHS PMID- 10807645 TI - Seven guidelines of wisdom PMID- 10807648 TI - Social inequalities need a different measure in men and women PMID- 10807647 TI - Inhaled corticosteroids reduce exacerbations in chronic obstructive pulmonary disease PMID- 10807649 TI - Remember that the treatment can be worse than the disease PMID- 10807650 TI - Head injury trials must be bigger and better PMID- 10807651 TI - Medical patients should be asked about ideas of suicide PMID- 10807652 TI - Parents prefer pads for collecting urine samples from their children PMID- 10807653 TI - Caspase inhibition and limitation of myocardial infarct size: protection against lethal reperfusion injury. AB - Ischaemia-reperfusion injury causes cell death by both necrosis and apoptosis. Caspase activation is a major event in apoptosis. We therefore examined the effect of caspase inhibitors during reperfusion upon myocardial infarction. Rat isolated hearts were subjected to 35 min coronary occlusion and 120 min reperfusion. Treatment groups were perfused with caspase inhibitors during early reperfusion. We assessed a non-selective caspase inhibitor (Z-VAD. fmk, 0.1 microM), a caspase-8 inhibitor (Z-IETD.fmk, 0.07 microM), a caspase-9 inhibitor (Z-LEHD.fmk, 0.07 microM) and a caspase-3 inhibitor (Ac-DEVD.cmk, 0.07 microM). All caspase inhibitors limited infarct size (infarct-risk ratio per cent: control 38.5+/-2.6; Z-VAD. fmk 24.6+/-3.4; Z-LEHD.fmk 19.3+/-2.4; Z-IETD.fmk 23.0+/-5.4; Ac-DEVD.cmk 27.8+/-3.3; P<0.05 when compared with control value, 1-way ANOVA). We conclude that caspase inhibition during early reperfusion protects myocardium against lethal reperfusion injury. PMID- 10807654 TI - Contractile responses to human urotensin-II in rat and human pulmonary arteries: effect of endothelial factors and chronic hypoxia in the rat. AB - Responses to human urotensin-II (hU-II) were investigated in human and rat pulmonary arteries. Rat pulmonary arteries: hU-II was a potent vasoconstrictor of main pulmonary arteries (2 - 3 mm i.d.) (pEC(50), 8.55+/-0.08, n=21) and was approximately 4 fold more potent than endothelin-1 [ET-1] (P<0.01), although its E(max) was considerably less (approximately 2.5 fold, P<0.001). The potency of hU II increased 2.5 fold with endothelium removal (P<0.05) and after raising vascular tone with ET-1 (P<0.01). E(max) was enhanced approximately 1.5 fold in the presence of N(omega)-nitro-L-arginine methylester (L-NAME, 100 microM, P<0.01) and approximately 2 fold in vessels from pulmonary hypertensive rats exposed to 2 weeks chronic hypoxia (P<0.05). hU-II did not constrict smaller pulmonary arteries. Human pulmonary arteries ( approximately 250 microm i.d.): in the presence of L-NAME, 3 out of 10 vessels contracted to hU-II and this contraction was highly variable. hU-II is, therefore, a potent vasoconstrictor of rat main pulmonary arteries and this response is increased by endothelial factors, vascular tone and onset of pulmonary hypertension. Inhibition of nitric oxide synthase uncovers contractile responses to hU-II in human pulmonary arteries. PMID- 10807655 TI - Acute blood pressure effects of YC-1-induced activation of soluble guanylyl cyclase in normotensive and hypertensive rats. AB - We used YC-1 as a pharmacological tool to investigate the short-term blood pressure effects of NO-independent activation of sGC in normotensive and hypertensive rats. Four groups of normotensive Wistar-Kyoto rats were treated by i.v. injection with vehicle (V), YC-1 (YC-1), sodium nitroprusside (SNP), or YC-1 and SNP (YC-1+SNP). Hypertension was induced in four additional groups of WKY rats by 3 weeks of oral treatment with L-NAME. These animals were investigated with the same protocol as the normotensive animals: L-NAME/V, L-NAME/YC-1, L NAME/SNP, L-NAME/YC-1+SNP. YC-1 lowered mean arterial blood pressure (MAP) in normotensive and hypertensive animals similarly to SNP alone (P<0.05, respectively). The combination of YC-1 with SNP caused a strong decrease of MAP in both the hypertensive and normotensive animals (P<0.05, respectively). SNP with YC-1 also induced a pronounced cyclic GMP increase in the aorta. This study shows for the first time the blood pressure lowering potential of bimodal targeting of the NO-sGC-system. PMID- 10807656 TI - Arginine and nitrate tolerance. PMID- 10807657 TI - Role of L-arginine in the vascular actions and development of tolerance to nitroglycerin. AB - The goal of this work was to test the role of nitric oxide synthase (NOS) and its substrate L-arginine in development of tolerance to nitroglycerin's (GTN) vasodilator actions. GTN's effects on NOS activity and NO formation were tested in cultured bovine aortic endothelial cells (BAECs). The arginine to citrulline conversion assay showed that GTN stimulated NOS basal activity in BAECs by approximately 40%, comparable with acetylcholine (ACh)-treated controls. Both effects were blocked by L-NMMA. Photometric assays showed that both GTN and ACh stimulated NO formation. Both effects were potentiated by L-arginine and inhibited by L-NAME. L-NAME inhibited ACh responses approximately 80% compared with approximately 40% for GTN responses. The aortic ring assay showed that 2 h pretreatment with GTN caused substantial tolerance to GTN's vasodilating effects as evidenced by a 38 fold rightward shift of the concentration-relaxation curve. In contrast to D-arginine, addition of L-arginine substantially inhibited this effect, reducing the rightward shift to 4.4 fold of control values. GTN tolerance was associated with a 40% reduction in L-arginine tissue levels. GTN had a biphasic effect on BAEC uptake of L-arginine, stimulating uptake at 5 and 15 min, and suppressing uptake after 1 and 4 h In summary, acute GTN treatment stimulates endothelial NOS activity in producing NO and increases cellular uptake of L arginine. Prolonged GTN exposure reduces GTN's vasodilator actions, decreases L arginine tissue levels and depresses BAECs uptake of L-arginine. Supplementation of L-arginine reduces development of GTN tolerance. These data indicate that GTN tolerance depends in part on activation of the NOS pathway. PMID- 10807658 TI - Attenuation of pressure-induced myogenic contraction and tyrosine phosphorylation by fasudil, a cerebral vasodilator, in rat cerebral artery. AB - The mechanism by which fasudil inhibits pressure-induced myogenic contraction was studied with regard to tyrosine phosphorylation in rat cerebral artery. Intracellular Ca(2+) concentration ([Ca(2+)](i)) and vessel diameter were simultaneously measured. Total tyrosine phosphorylation level and phosphorylation of tyrosine 419 on pp60(src) required for its full catalytic activity were immunocytochemically detected in situ. Fasudil (1 - 100 microM) partially suppressed the increase in [Ca(2+)](i), and totally attenuated contraction elicited by pressurization from 10 to 60 mmHg. Furthermore, fasudil (100 microM) significantly attenuated tyrosine phosphorylation and the activity of pp60(src) augmented in situ by pressure. Herbimycin A (1 - 100 nM) and genistein (3 - 30 microM), tyrosine kinase inhibitors, effectively attenuated the pressure-induced increase in [Ca(2+)](i), contraction, tyrosine phosphorylation, and activation of pp60(src). Both fasudil and herbimycin A directly inhibited the pp60(src) activity in a cell free system. Orthovanadate (100 microM), a tyrosine phosphatase inhibitor, significantly potentiated the pressure-induced increase in [Ca(2+)](i) and contraction. Nicardipine (100 nM), a Ca(2+) antagonist, completely inhibited pressure-induced increase in [Ca(2+)](i) and contraction, but affected neither tyrosine phosphorylation nor activity of pp60(src) in the pressurized arteries. Arginine-glycine-aspartic acid-serine peptide (1 - 100 microM) concentration-dependently reduced the pressure-induced contraction. In addition to the hitherto reported vasodilatory actions of fasudil, the present results suggest the inhibition by fasudil of pressure-induced tyrosine phosphorylation and pp60(src) activation. The wide spectrum of inhibitory actions of fasudil may contribute to the effective attenuation of the pressure-induced contraction in the cerebral artery. PMID- 10807659 TI - Suppression of arterial intimal hyperplasia by cilostamide, a cyclic nucleotide phosphodiesterase 3 inhibitor, in a rat balloon double-injury model. AB - The effects of cilostamide, a cyclic nucleotide phosphodiesterase 3 (PDE3) selective inhibitor, on vascular intimal hyperplasia were evaluated using a single-balloon injury model and a double-injury model in which the rat common carotid artery was subjected to a second injury at a site injured 14 days previously. In the double-injury model, the second balloon injury caused more severe intimal hyperplasia (intima/media (IM) ratio, 1.88+/-0.10) than in the single-injury model (1.09+/-0.08). Histopathological study revealed that vascular smooth muscle cells (VSMC) were the predominant cell-type in the affected neointimal area. Oral administration of cilostamide for 2 weeks after the second injury suppressed intimal hyperplasia in the double-injury model (30 mg kg(-1) bid, 83% inhibition in terms of the IM ratio, P<0.05; 100 mg kg(-1) bid, 69% inhibition, P<0.05). Similar effects were also observed in the single-injury model with oral administration of cilostamide for 2 weeks (100 mg kg(-1) bid, 36% inhibition, P<0.01). Cilostamide inhibited DNA synthesis of cultured VSMC stimulated by foetal calf serum or different kinds of growth factors, but did not affect their migration stimulated by platelet-derived growth factor (PDGF)-BB. Cilostamide significantly increased the cyclic AMP concentration of VSMC dose dependently. These results indicate that cilostamide suppresses intimal hyperplasia both in the single- and double-injury models of rat, presumably by inhibiting proliferation rather than migration of VSMC. It is suggested that PDE3 inhibitors might find application in preventing intimal hyperplasia following angioplasty such as percutaneous transluminal coronary angioplasty (PTCA) or stent. PMID- 10807660 TI - Comparison of serum, cerebrospinal fluid and brain extracellular fluid pharmacokinetics of lamotrigine. AB - We investigated the rate of penetration into and the intra-relationship between the serum, cerebrospinal fluid (CSF) and regional brain extracellular fluid (bECF) compartments following systemic administration of lamotrigine in rat. The serum pharmacokinetics were biphasic with an initial distribution phase, (half life approximately 3 h), and then a prolonged elimination phase of over 30 h. The serum pharmacokinetics were linear over the range 10 - 40 mg kg(-1). Using direct sampling of CSF with concomitant serum sampling, the calculated penetration half time into CSF was 0.42+/-0.15 h. At equilibrium, the CSF to total serum concentration ratio (0.61+/-0.02) was greater than the free to total serum concentration (0.39+/-0.01). Using in vivo recovery corrected microdialysis sampling in frontal cortex and hippocampus with concomitant serum sampling, the calculated penetration half-time of lamotrigine into bECF, 0.51+/-0.11 h, was similar to that for CSF and was not area or dose dependent. At equilibrium, the bECF to total serum concentration ratio (0.40+/-0.04) was similar to the free to total serum concentration (0.39+/-0.01), and did not differ between hippocampus and frontal cortex. The species specific serum kinetics can explain the prolonged action of lamotrigine in rat seizure models. Lamotrigine has a relatively slow penetration into both CSF and bECF compartments compared with antiepileptic drugs used in acute seizures. Furthermore, the free serum drug concentration is not the sole contributor to the CSF compartment, and the CSF concentration is an overestimate of the bECF concentration of lamotrigine. PMID- 10807661 TI - Activation and block of recombinant GABA(A) receptors by pentobarbitone: a single channel study. AB - Recombinant GABA(A) receptors (alpha1beta2gamma2L) were transiently expressed in HEK 293 cells. We have investigated activation and block of these receptors by pentobarbitone (PB) using cell-attached single-channel patch clamp. Clusters of single-channel activity elicited by 500 microM PB were analysed to estimate rate constants for agonist binding and channel gating. The minimal model able to describe the kinetic data involved two sequential binding steps, followed by channel opening. The estimated channel opening rate constant is approximately 1500 s(-1), and the estimated equilibrium dissociation constants for the binding steps involved in activation are approximately 2 mM. Our results show a dose dependent block of receptors at millimolar concentrations of PB that results in reduced open interval durations. The reduction in mean open time is linearly proportional to PB concentration, indicating that block can be produced by binding of a single PB molecule. Addition of millimolar concentrations of PB in the presence of GABA also produces a reduction of open channel lifetime in addition to a progressive increase in the closed interval durations within a cluster. The data suggest that the receptor contains two or more blocking sites while occupancy of only one of the sites is sufficient for channel block. Neither the blocking rate constant nor return rate from the blocked state(s) is affected by pH (ionization status of the PB molecule) demonstrating that both neutral and anionic forms of PB cause channel block. PMID- 10807663 TI - Non-NF-kappaB elements are required for full induction of the rat type II nitric oxide synthase in vascular smooth muscle cells. AB - We have investigated the role of the NF-kappaB binding sites and other promoter elements beyond NF-kappaB in iNOS induction in rat vascular smooth muscle cells (SMC). Rat aortic SMC transfected with iNOS promoter constructs with either mutation or deletion of the downstream NF-kappaB site exhibited about 50% reduction in promoter activity in response to a cytokine mixture, whereas either mutation or deletion of the upstream NF-kappaB site reduced promoter activity by 90%, suggesting that the latter site is the most important, and that co-existence of two NF-kappaB sites is necessary for iNOS induction. Nuclear NF-kappaB activity was robustly induced by TNF-alpha. However, TNF-alpha alone did not induce iNOS promoter activity, protein expression, or nitrite production, indicating that NF-kappaB activation alone is not sufficient for iNOS induction. The construct up to -890 bp, containing the downstream NF-kappaB site, exhibited little response to cytokines. The construct up to -1.0 kb, containing the two NF kappaB sites exhibited only 22% of full promoter activity. The regions -1001 to 1368 bp and -2 to -2.5 kb contributed an additional 43 and 22% promoter activity, respectively. Internal deletion or reversal of the orientation of -1001 to -1368 bp in the full promoter resulted in 40% reduction in promoter activity. These data suggest that the co-existence of two NF-kappaB sites is essential for core promoter activity, but that full induction of the rat SMC iNOS gene requires other elements located between -1.0 to -1.37 and -2.0 to -2.5 kb of the promoter. PMID- 10807662 TI - Inhibition by adenosine A(2A) receptors of NMDA but not AMPA currents in rat neostriatal neurons. AB - Whole-cell patch clamp experiments were used to investigate the transduction mechanism of adenosine A(2A) receptors in modulating N-methyl-D-aspartate (NMDA) induced currents in rat striatal brain slices. The A(2A) receptor agonist 2-p-(2 carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS 21680) inhibited the NMDA, but not the (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) current in a subset of striatal neurons. Lucifer yellow-filled pipettes in combination with immunostaining of A(2A) receptors were used to identify CGS 21680-sensitive cells as typical medium spiny striatal neurons. Dibutyryl cyclic AMP and the protein kinase A activator Sp-cyclic AMPs, but not the protein kinase A inhibitors Rp-cyclic AMPS or PKI(14 - 24)amide abolished the inhibitory effect of CGS 21680. The phospholipase C inhibitor U-73122, but not the inactive structural analogue U-73343 also interfered with CGS 21680. The activation of protein kinase C by phorbol 12-myristate 13-acetate or the blockade of this enzyme by staurosporine did not alter the effect of CGS 21680. Heparin, an antagonist of inositol 1, 4,5-trisphosphate (InsP(3)) and a more efficient buffering of intracellular Ca(2+) by BAPTA instead of EGTA in the pipette solution, abolished the CGS 21680-induced inhibition. The calmodulin antagonist W 7 and cytochalasin B which enhances actin depolymerization also prevented the effect of CGS 21680; the calmodulin kinase II inhibitors CaM kinase II(281 - 309) and KN-93 but not the inactive structural analogue KN-92 were also effective. The calcineurin inhibitor deltamethrin did not interfere with CGS 21680. It is suggested that the transduction mechanism of A(2A) receptors to inhibit NMDA receptor channels is the phospholipase C/InsP(3)/calmodulin and calmodulin kinase II pathway. The adenylate cyclase/protein kinase A and phospholipase C/protein kinase C pathways do not appear to be involved. PMID- 10807664 TI - GABA(B) receptor-mediated effects on vagal pathways to the lower oesophageal sphincter and heart. AB - GABA(B) receptors influencing vagal pathways to the lower oesophageal sphincter and heart were investigated. In urethane-anaesthetized ferrets, the GABA(B) agonist baclofen (7 micromol kg(-1) i.v.) increased basal lower oesophageal sphincter (LOS) pressure. This was reversed by antagonism with CGP35348 (100 micromol kg(-1) i.v.). Baclofen's effect was abolished by vagotomy, suggesting a central action, yet it was ineffective when given centrally (3 - 6 nmol i.c.v.). Peripheral vagal stimulation (10 Hz, 5 s duration) caused LOS inhibition, followed by excitation, then prolonged inhibition. Bradycardia was also evoked during stimulation. Bradycardia and LOS responses were abolished after chronic supranodose vagotomy, indicating that they were due to stimulation of vagal pre ganglionic neurones, not antidromic stimulation of afferents. Baclofen (1 - 10 micromol kg(-1)) reduced bradycardia and enhanced LOS excitation, which was also seen in animals pretreated with atropine (400 microgram kg(-1) i.v.) and guanethidine (5 mg kg(-1) i.v.), but not in those pretreated with L-NAME (100 mg kg(-1) i.v.). Effects of baclofen (7 micromol kg(-1) i.v.) on vagal stimulation induced LOS and cardiac responses were unchanged by the GABA(B) antagonists CGP35348 or CGP36742 (up to 112 micromol kg(-1) i.v.), but were reversed by CGP62349 (ED(50) 37 nmol kg(-1) i.v.) or CGP54626 (ED(50) 100 nmol kg(-1) i.v.). Responses of isolated LOS strips to electrical stimulation, capsaicin, NK-1, NK-2 and nicotinic receptor agonists were all unaffected by baclofen (90% of total binding and was fully reversible by methiothepin (10 microM), full dissociation occurring by 100 min. The association (k(+1)) and dissociation (k( 1)) rate constants were 0.05 nM(-1)min(-1) and 0.05 min(-1) respectively, giving a K(D) (k(-1)/k(+1)) of 1.0 nM. [(3)H]-SB-269970 bound saturably and apparently monophasically to both h5-HT(7(a))/293 and guinea-pig cortex membranes, with K(D) values of 1.25+/-0.05 and 1.7+/-0.3 nM respectively. The B(max) for [(3)H]-SB 269970 to both h5-HT(7(a))/293 and guinea-pig cortex membranes (5780+/-380 and 125+/-8.2 fmoles mg protein(-1) respectively) was similar to that for [(3)H]-5-CT (6190+/-940 and 143+/-19 fmoles mg protein(-1) respectively). These data suggest that, in each tissue, both radioligands labelled the same population of receptors, which appear to be present in an agonist high affinity state. The profile of compound inhibition of [(3)H]-SB-269970 binding to h5-HT(7(a))/293 and guineapig cortex membranes correlated well (corr. coeff. 0.98) with those for [(3)H]-5-CT binding and were consistent with the profiles reported previously for the human 5-HT(7(a)) and guinea-pig cortex 5-HT(7) receptors using [(3)H]-5-CT. Hill slopes for inhibition of [(3)H]-SB-269970 and [(3)H]-5-CT binding were close to 1, consistent with binding to a single receptor population in both tissues. [(3)H]-SB-269970 represents the first selective 5-HT(7) antagonist radioligand, which should aid further characterization of 5-HT(7) receptors in recombinant and native tissues and help establish their role in brain function. PMID- 10807681 TI - The phosphatidylinositol 4-kinase inhibitor phenylarsine oxide blocks evoked neurotransmitter release by reducing calcium entry through N-type calcium channels. AB - The effects of the phosphatidylinositol 4-kinase inhibitor, phenylarsine oxide (PAO), on acetylcholine (ACh) release and on prejunctional Ca(2+) currents were studied at the frog neuromuscular junction using electrophysiological recording techniques. Application of PAO (30 microM) increased both spontaneous ACh release reflected as miniature end-plate potential (mepp) frequencies and evoked ACh release reflected as end-plate potential (epp) amplitudes with a similar time course. Following the initial increase in epp amplitudes produced by PAO, epps slowly declined and were eventually abolished after approximately 20 min. However, mepp frequencies remained elevated over this time period. PAO (30 microM) also inhibited the perineural voltage change associated with Ca(2+) currents through N-type Ca(2+) channels (prejunctional Ca(2+) currents) at motor nerve endings. Addition of British anti-lewisite (BAL, 1 mM), an inactivator of PAO, partially reversed both the inhibition of epps and the inhibition of the prejunctional Ca(2+) current. The effects of PAO on N-type Ca(2+) channels were investigated more directly using the whole cell patch clamp technique on acutely dissociated sympathetic neurons. Application of PAO (30 - 40 microM) to these neurons decreased the voltage-activated calcium currents through N-type Ca(2+) channels, an effect that was partially reversible by BAL. In combination, these results suggest that inhibition of neurotransmitter release by PAO occurs as a consequence of the inhibition of Ca(2+) entry via N-type calcium channels. The relationship between the effects of PAO on N-type Ca(2+) channels in motor nerve endings and in neuronal soma is discussed. PMID- 10807682 TI - Modulation of 5-hydroxytryptamine efflux from rat cortical synaptosomes by opioids and nociceptin. AB - The modulation of [(3)H]-5-hydroxytryptamine ([(3)H]-5-HT) efflux from superfused rat cortical synaptosomes by delta, kappa, mu and ORL(1) opioid receptor agonists and antagonists was studied. Spontaneous [(3)H]-5-HT efflux was reduced (20% inhibition) by either 0.5 microM tetrodotoxin or Ca(2+)-omission. Ten mM K(+) evoked [(3)H]-5-HT overflow was largely Ca(2+)-dependent (90%) and tetrodotoxin sensitive (50%). The delta receptor agonist, deltorphin-I, failed to modulate the K(+)-evoked neurotransmitter efflux up to 0.3 microM. The kappa and the mu receptor agonists, U-50,488 and endomorphin-1, inhibited K(+)-evoked [(3)H]-5-HT overflow (EC(50)=112 and 7 nM, respectively; E(max)=28 and 29% inhibition, respectively) in a norBinaltorphimine- (0.3 microM) and naloxone- (1 microM) sensitive manner, respectively. None of these agonists significantly affected spontaneous [(3)H]-5-HT efflux. The ORL(1) receptor agonist nociceptin inhibited both spontaneous (EC(50)=67 nM) and K(+)-evoked (EC(50)=13 nM; E(max)=52% inhibition) [(3)H]-5-HT efflux. The effect of NC was insensitive to naloxone (up to 10 microM), but was antagonized by [Nphe(1)]nociceptin(1-13)NH(2) (a novel selective ORL(1) receptor antagonist; pA(2)=6.7) and by naloxone benzoylhydrazone (pA(2)=6.3). The ORL(1) ligand [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) also inhibited K(+) stimulated [(3)H]-5-HT overflow (EC(50)=64 nM; E(max)=31% inhibition), but its effect was partially antagonized by 10 microM naloxone. It is concluded that the ORL(1) receptor is the most important presynaptic modulator of neocortical 5-HT release within the opioid receptor family. This suggests that the ORL(1)/nociceptin system may have a powerful role in the control of cerebral 5-HT-mediated biological functions. PMID- 10807683 TI - Modulation of the dinucleotide receptor present in rat midbrain synaptosomes by adenosine and ATP. AB - Diadenosine polyphosphates activate dinucleotide receptors in rat midbrain synaptic terminals. The agonist with highest affinity at this receptor, diadenosine pentaphosphate (Ap(5)A), elicits Ca(2+) transients at concentrations ranging from 10(-7) to 10(-3) M with a single-phase curve and an EC(50) value of 56.21+/-1.82 microM. Treatment of synaptosomal preparations with alkaline phosphatase (AP) changes the dose-response control curve into a biphasic one presenting two EC(50) values of 6.47+/-1.25 nM and 11.16+/-0.83 microM respectively. The adenosine A(1) antagonist 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX) reversed the biphasic concentration-response for Ap(5)A curve in the presence of AP, to a monophasic one with an EC(50) value of 76.05+/-7.51 microM. The application of adenosine deaminase produced the same effect as DPCPX, the EC(50) value for Ap(5)A, in the presence of AP being 18.62+/-4.03 microM. Activation of the adenosine A(1) receptor by means of cyclohexyladenosine (CHA) shifted the dose response curve for Ap(5)A to the left, resulting in a monophasic curve with an EC(50) of 5. 01+/-0.02 pM. The destruction of extrasynaptosomal nucleotides by AP or the addition of pyridoxalphosphate-6-azophenyl-2',4' disulphonic acid (PPADS), a broad P2 antagonist compound, enhance maximal effect of the Ap(5)A up to 55.6% on the dose response curve, thus suggesting a negative modulation by P2 receptors. In a summary, ATP and adenosine present at the extra synaptosomal space, are relevant natural modulators of the dinucleotide receptor, via P2 and adenosine A(1) receptors respectively. PMID- 10807684 TI - Localization of the nephron site of gentamicin-induced hypercalciuria in the rat: a micropuncture study. AB - In vivo renal micropuncture techniques were used to locate the nephron site of hypercalciuria induced by acute gentamicin infusion in anaesthetized Sprague Dawley rats. Three series of experiments were conducted. The effect of gentamicin on calcium reabsorption in the proximal tubule (Series I) and loop of Henle (Series II) was investigated using in vivo microperfusion whereas the effect on distal calcium handling (Series III) was studied using in vivo microinfusion. In all three experimental series, acute systemic gentamicin infusion at 0.28 mg kg( 1) min(-1) caused significant hypercalciuria within 30 min of commencing drug infusion. Gentamicin had no effect on the rates of urine flow or sodium excretion. Acute gentamicin infusion had no effect on unidirectional calcium reabsorption in the proximal tubule or loop of Henle despite a simultaneous and highly significant hypercalciuria at the whole kidney level. Net fluid reabsorption was also unaffected by the drug in these nephron segments. Acute gentamicin infusion significantly increased the urinary recovery of calcium following microinfusion into early distal tubules, whereas urinary calcium recovery was decreased after microinfusion into late distal tubules. We conclude that acute gentamicin-induced hypercalciuria is mediated by a decrease in calcium reabsorption in the early distal tubule. Thus, the acute hypercalciuric effect of gentamicin occurs at a different nephron site to the nephrotoxic effects associated with longer-term administration of the drug. It is, therefore, unlikely that gentamicin-induced hypercalciuria is involved in the pathogenesis of subsequent proximal tubular cell injury. PMID- 10807685 TI - Pentaerythrityl tetranitrate attenuates structural changes in conduit arteries evoked by long-term NO-synthase inhibition. AB - The aim of the study was to determine whether the pentaerythrityl tetranitrate (PETN), a tolerance devoid exogenous NO donor could prevent morphological changes in the cardiovascular system evoked by long-term NO-synthase inhibition. Three groups of 10-week-old Wistar rats were used: (1) controls, (2) treated by L-N(G) nitroarginine methyl ester (L-NAME) in water (50 mg kg(-1)), and (3) treated by L NAME (50 mg kg(-1) in water)+PETN (2x50 mg kg(-1), using gavage). Blood pressure (BP) was measured by the tail plethysmographic method. After sacrificing the animals were perfused (120 mmHg) by glutaraldehyde fixative and processed according to standard electron microscopy procedure. Wall thickness (WT), cross sectional area (CSA), inner diameter (ID) of thoracic aorta (TA), carotid (CA) and septal branch of the left descending coronary artery (RS) were measured in light microscopy. After 6 weeks, the BP was increased to 172+/-1.7 mmHg (P<0.01) in the L-NAME group, compared to 127+/-1.4 mmHg in controls. In L-NAME+PETN treated rats, BP was 163+/-0.9 mmHg (P<0.01), and significantly lower (P<0.01) in comparison to L-NAME-treated rats. Heart weight and heart/body weight ratio was not significantly changed. In L-NAME-treated rats, both WT and CSA were increased in all three arteries (P<0.01). ID was increased only in TA (P<0.01). Wall/diameter ratio (WD) was increased in TA (P<0. 01) and CA (P<0.01). In L NAME+PETN treated rats, WT was found to be increased only in TA (P<0.01). In comparison to the L-NAME treated group, WT was decreased in TA (P<0.01), in CA (P<0.01), in RS (P<0. 05). CSA was increased only in TA (P<0.01), yet in comparison to the L-NAME group it was decreased in CA (P<0.01). ID was increased in comparison to both control and L-NAME treated rats only in TA (P<0. 01). WD did not differ from the control value. In comparison to L-NAME-treated rats, it was decreased in both TA and CA (P<0.01). These data suggest that the changes in the cardiovascular system evoked by long-term NO-synthase inhibition were attenuated by simultaneous administration of the exogenous donor of nitric oxide PETN. PMID- 10807686 TI - Inhibition of pulmonary eosinophilia and airway hyperresponsiveness in allergic mice by rolipram: involvement of endogenously released corticosterone and catecholamines. AB - This study investigates the role of adrenal-derived catecholamines and corticosterone on the inhibition by rolipram, a phosphodiesterase (PDE)-4 inhibitor, of pulmonary eosinophilia and airway hyperresponsiveness (AHR) in allergic mice. The following experimental groups were studied in mice sensitized and challenged with ovalbumin (OVA): normal, adrenalectomized, propranolol (beta adrenoceptor antagonist) and metyrapone (corticosterone synthesis inhibitor) treated. These interventions were studied both in the absence and in the presence of rolipram. Eosinophil numbers in the bronchoalveolar lavage (BAL) and AHR to methacholine were measured 24 h after OVA challenge. Treatment of sensitized mice with rolipram (0.3 - 10 mg kg(-1), p.o.), inhibited pulmonary eosinophilia and the AHR to methacholine in OVA-challenged mice. Adrenalectomy increased the number of eosinophils in the BAL of OVA-challenged mice but had no effect on AHR to methacholine. Adrenalectomy attenuated both the rolipram-induced inhibition of BAL eosinophilia and AHR to methacholine in OVA challenged mice. Propranolol (10 mg kg(-1), p.o.) had no effect on the inhibition of eosinophilia by rolipram but attenuated the inhibition of AHR to methacholine in OVA challenged mice. On the other hand, metyrapone (10 mg kg(-1), p.o.) attenuated the inhibition of eosinophilia by rolipram but had no effect on the inhibition of AHR to methacholine in OVA challenged mice. Metyrapone-treatment alone increased the number of eosinophils in the BAL of OVA-challenged mice. These results identify an important role for adrenal-derived catecholamines and corticosterone on the inhibition of pulmonary eosinophilia and AHR by rolipram in allergic mice. PMID- 10807687 TI - CGRP(2) receptor in the internal anal sphincter of the rat: implications for CGRP receptor classification. AB - The CGRP receptor mediating relaxation of the rat internal anal sphincter (IAS) has been characterized using CGRP analogues, homologues, the antagonist CGRP(8 - 37) and its analogues. In isolated IAS strips, the spontaneously developed tone was concentration-dependently relaxed by halpha CGRP, hbeta CGRP and rat beta CGRP (pEC(50) 8.1+/-0.2, 8.3+/-0.1 and 8.4+/-0.2, respectively; 100% maximum response). Vasoactive intestinal polypeptide (VIP) was around 7 fold more potent than halpha CGRP (pEC(50) 9.0+/-0.1; 100% maximum relaxation). [Cys(ACM(2.7))] halpha CGRP and salmon calcitonin were inactive (up to 10(-5) M). Halpha CGRP(8 - 37) (10(-5) M) antagonized responses to halpha CGRP (apparent pK(B) 5.7+/-0.3) and rat beta CGRP (apparent pK(B) 5.8+/-0.2), but not to VIP. Hbeta CGRP(8 - 37) (10(-5) M) was an antagonist against halpha CGRP (apparent pK(B) 6.1+/-0.1). Halpha CGRP(8 - 37) analogues (10(-5) M), with substitutions at the N-terminus by either glycine(8) or des-NH(2) valine(8) or proline(8), antagonized halpha CGRP responses with similar affinities (apparent pK(B) 5.8+/-0.1, 5.8+/-0.1 and 5.5+/ 0.1, respectively). Peptidase inhibitors (amastatin, bestatin, captopril, phosphoramidon and thiorphan, 10(-6) M each) did not increase the agonist potency of either halpha CGRP or [Cys(ACM(2,7))] halpha CGRP, or the antagonist affinity of halpha CGRP(8 - 37) against halpha CGRP or rat beta CGRP. These data demonstrate for the first time a CGRP receptor in the rat IAS for which halpha CGRP (8 - 37) and its analogues have an affinity that is consistent with a CGRP(2) receptor. However, there is a marked species difference as the antagonist has a 100 fold lower affinity in the rat than in the same tissue of the opossum (Chakder & Rattan, 1991). PMID- 10807688 TI - Thyrotropin regulates adenosine A(1) receptor expression in rat thyroid FRTL-5 cells. AB - The effect of thyrotropin (TSH), on adenosine A(1) receptor expression in thyroid FRTL-5 cells was examined by [(3)H]-1, 3-dipropyl-,8-cyclopentyl xanthine ([(3)H] DPCPX) binding on cells in suspension and on membrane preparation, and by in situ mRNA labelling. The estimated K(D) for intact cells was 0.19 nM and about 47,000 binding sites per cell were found in cells constantly grown in the presence of TSH. Three days deprivation of TSH decreased the number of [(3)H]-DPCPX binding sites without any significant effect of K(D). Reintroduction of TSH to the cells returned the higher level of A(1) receptors both in suspension binding studies on whole cells and on membrane preparations. In situ hybridization revealed that TSH evoked an increase in the number of cells densely labelled with a probe against A(1) receptor mRNA. The potency of the A(1) receptor agonist N(6) cyclohexyladenosine (CHA) as an inhibitor of cyclic AMP formation induced by forskolin was increased in TSH-treated cells, with a shift in the IC(50) from 2.05 nM in TSH-deprived cells to 0.14 nM in TSH-treated cells. Since the activation of A(1) receptors inhibits TSH-mediated cyclic AMP signalling, our results suggest a regulatory feedback mechanism between signalling via adenosine A(1) receptors and TSH receptors. PMID- 10807689 TI - Induced desensitization of the insulinotropic effects of antidiabetic drugs, BTS 67 582 and tolbutamide. AB - Acute and chronic mechanisms of action of novel insulinotropic antidiabetic drug, BTS 67 582 (1, 1-dimethyl-2-(2-morpholinophenyl)guanidine fumarate), were examined in the stable cultured BRIN-BD11 cell line. BTS 67 582 (100 - 400 microM) stimulated a concentration-dependent increase (P<0.01) in insulin release at both non-stimulatory (1.1 mM) and stimulatory (8. 4 mM) glucose. Long-term exposure (3 - 18 h) to 100 microM BTS 67 582 in culture time-dependently decreased subsequent responsiveness to acute challenge with 200 microM BTS 67 582 or 200 microM tolbutamide at 12 - 18 h (P<0.001). Similarly 3 - 18 h culture with the sulphonylurea, tolbutamide (100 microM), also effectively suppressed subsequent insulinotropic responses to both BTS 67 582 and tolbutamide. Culture with 100 microM BTS 67 582 or 100 microM tolbutamide did not affect basal insulin secretion, cellular insulin content, or cell viability and exerted no influence on the secretory responsiveness to 200 microM of the imidazoline, efaroxan. While 18 h BTS 67 582 culture did not affect the insulin-releasing actions (P<0.001) of 16.7 mM glucose, 10 mM arginine, 30 mM KCl, 25 microM forskolin or 10 nM phorbol 12-myristate 13-acetate (PMA), significant inhibition (P<0.001) of the insulinotropic effects of 10 mM 2-ketoisocaproic acid (KIC) and 10 mM alanine were observed. These data suggest that BTS 67 582 shares a common signalling pathway to sulphonylurea but not imidazoline drugs. Desensitization of drug action may provide an important approach to dissect sites of action of novel and established insulinotropic antidiabetic agents. PMID- 10807691 TI - Novel germline mutations in the PTEN tumour suppressor gene found in women with multiple cancers. AB - Germline mutations in PTEN can predispose people to Cowden syndrome (CS) and Bannayan-Ruvalcaba-Riley (BRR) syndrome, rare, autosomal dominantly inherited neoplastic disorders. To determine whether germline mutations in PTEN contribute to genetic predisposition to multiple primary tumours within the general population, we conducted a nested case-control study, among 32 826 members of the prospective Nurses' Health Study cohort; cases were women with more than one primary tumour at different anatomical sites. We screened all nine exons of PTEN and flanking intronic splice sites for all 103 eligible cases using SSCP and sequencing. We observed two novel germline heterozygous missense mutations in exon 5 in five of the cases; three were V119L and two were V158L. Neither mutation was observed in 115 controls free of diagnosed cancer (p = 0.02). Both mutants showed partial tumour suppressor activity when compared to wild type PTEN when transfected into a PTEN null breast cancer cell line. The phenotype was cell line specific suggesting that genetic background affects growth suppression activity of the mutants. These data provide evidence that germline mutations in PTEN may be a more frequent predisposing factor for cancers in women than previously suggested. PMID- 10807690 TI - The Smith-Lemli-Opitz syndrome. AB - The Smith-Lemli-Opitz syndrome (SLOS) is one of the archetypical multiple congenital malformation syndromes. The recent discovery of the biochemical cause of SLOS and the subsequent redefinition of SLOS as an inborn error of cholesterol metabolism have led to important new treatment possibilities for affected patients. Moreover, the recent recognition of the important role of cholesterol in vertebrate embryogenesis, especially with regard to the hedgehog embryonic signalling pathway and its effects on the expression of homeobox genes, has provided an explanation for the abnormal morphogenesis in the syndrome. The well known role of cholesterol in the formation of steroid hormones has also provided a possible explanation for the abnormal behavioural characteristics of SLOS. PMID- 10807692 TI - Chromosome 8p alterations in sporadic and BRCA2 999del5 linked breast cancer. AB - Chromosomal losses involving the short arm of chromosome 8 are frequent in a variety of tumour types, including breast cancer, suggesting the presence of one or more tumour suppressor genes in this region. In this study, we have used 11 microsatellite markers to analyse loss of heterozygosity (LOH) at chromosome 8p in 151 sporadic breast tumours and 50 tumours from subjects carrying the BRCA2 999del5 mutation. Fifty percent of sporadic tumours compared to 78% of BRCA2 linked tumours exhibit LOH at one or more markers at 8p showing that chromosome 8p alterations in breast tumours from BRCA2 999del5 carriers are more pronounced than in sporadic breast tumours. The pattern of LOH is different in the two groups and a higher proportion of BRCA2 tumours have LOH in a large region of chromosome 8p. In the total patient material, LOH of 8p is associated with LOH at other chromosome regions, for example, 1p, 3p, 6q, 7q, 9p, 11p, 13q, 17p, and 20q, but no association is found between LOH at 8p and chromosome regions 11q, 16q, 17q, and 18q. Furthermore, an association is detected between LOH at 8p and positive node status, large tumour size, aneuploidy, and high S phase fraction. Breast cancer patients with LOH at chromosome 8p have a worse prognosis than patients without this defect. Multivariate analysis suggests that LOH at 8p is an independent prognostic factor. We conclude that chromosome 8p carries a tumour suppressor gene or genes, the loss of which results in growth advantage of breast tumour cells, especially in carriers of the BRCA2 999del5 mutation. PMID- 10807693 TI - Familial clear cell renal cell carcinoma (FCRC): clinical features and mutation analysis of the VHL, MET, and CUL2 candidate genes. AB - Familial renal cell carcinoma (RCC) is genetically heterogeneous. Genetic predisposition to clear cell RCC (CCRCC) is a major feature of von Hippel-Lindau (VHL) disease (MIM 193300) and has rarely been associated with chromosome 3 translocations. In addition, familial papillary (non-clear cell) RCC may result from germline mutations in the MET proto-oncogene (MIM 164860). However, rare kindreds with familial CCRCC (FCRC) not linked to the VHL tumour suppressor gene have been described suggesting that further familial RCC susceptibility genes exist. To investigate the genetic epidemiology of FCRC, we undertook a clinical and molecular study of FCRC in nine kindreds with two or more cases of CCRCC in first degree relatives. FCRC was characterised by an earlier age at onset (mean 47.1 years, 52% of cases <50 years of age) than sporadic cases. These findings differ from the only previous report of two FCRC kindreds and have important implications for renal surveillance in FCRC. The molecular basis of CCRCC susceptibility was investigated in nine FCRC kindreds and seven isolated cases with features of possible genetic susceptibility to CCRCC (four bilateral CCRCC aged <50 years and three with unilateral CCRCC aged <30 years). No germline mutations were detected in the VHL or MET genes, suggesting that FCRC is not allelic with VHL disease or HPRC. As binding of the VHL gene product to the CUL2 protein is important for pVHL function, we then searched for germline CUL2 mutations. Although CUL2 polymorphisms were identified, no pathogenic mutations were detected. These findings further define the clinical features of FCRC and exclude a major role for mutations in VHL, MET, or CUL2 in this disorder. PMID- 10807694 TI - Efficacy of a touchscreen computer based family cancer history questionnaire and subsequent cancer risk assessment. AB - OBJECTIVE: A computer based touchscreen family cancer history questionnaire was developed and implemented to facilitate the provision of cancer risk assessments for the ambulatory and outpatient populations of a free standing cancer hospital. METHODS: A questionnaire consisting of a series of branched point decision making screens was developed which enables the participant to enter demographic data, personal cancer history, and cancer histories for first and second degree relatives. A freestanding touchscreen computer kiosk system was used to place the questionnaire in public areas of the cancer hospital and clinic. Genetic professionals analysed the data received, using published criteria, and provided a basic cancer risk assessment and surveillance recommendations within 10 business days. A survey was completed by a small random group of users (n = 59) three to six months after receipt of their risk assessment. RESULTS: After 11 months, 1440 people had entered information and received a written communication. Only 2% of completed questionnaires contained insufficient information to provide a basic risk assessment. Of the small group of participants surveyed, almost all (95%) felt "very comfortable" using the system, 93% remembered receiving the risk assessment letter when queried three to six months later, 42% felt their perceptions about cancer risk had changed, and 20% had made changes in their or their family's cancer surveillance practices. CONCLUSION: The touchscreen computer family history questionnaire allows easy collection of family history information, provision of risk assessments to a broad population, and promotes increased awareness of familial risk and appropriate surveillance. PMID- 10807696 TI - A novel C202F mutation in the connexin26 gene (GJB2) associated with autosomal dominant isolated hearing loss. AB - Mutations in the GJB2 gene encoding connexin26 (CX26) account for up to 50% of cases of autosomal recessive hearing loss. In contrast, only one GJB2 mutation has been reported to date in an autosomal dominant form of isolated prelingual hearing loss. We report here a novel heterozygous 605G-->T mutation in GJB2 in all affected members of a large family with late childhood onset of autosomal dominant isolated hearing loss. The resulting C202F substitution, which lies in the fourth (M4) transmembrane domain of CX26, may impair connexin oligomerisation. Finally, our study suggests that GJB2 should be screened for heterozygous mutations in patients with autosomal dominant isolated hearing impairment, whatever the severity of the disease. PMID- 10807695 TI - A cross section of autosomal recessive limb-girdle muscular dystrophies in 38 families. AB - Limb-girdle muscular dystrophies constitute a broad range of clinical and genetic entities. We have evaluated 38 autosomal recessive limb-girdle muscular dystrophy (LGMD2) families by linkage analysis for the known loci of LGMD2A-F and protein studies using immunofluorescence and western blotting of the sarcoglycan complex. One index case in each family was investigated thoroughly. The age of onset and the current ages were between 11/2 and 15 years and 6 and 36 years, respectively. The classification of families was as follows: calpainopathy 7, dysferlinopathy 3, alpha sarcoglycan deficiency 2, beta sarcoglycan deficiency 7, gamma sarcoglycan deficiency 5, delta sarcoglycan deficiency 1, and merosinopathy 2. There were two families showing an Emery-Dreifuss phenotype and nine showing no linkage to the LGMD2A-F loci, and they had preserved sarcoglycans. gamma sarcoglycan deficiency seems to be the most severe group as a whole, whereas dysferlinopathy is the mildest. Interfamilial variation was not uncommon. Cardiomyopathy was not present in any of the families. In sarcoglycan deficiencies, sarcoglycans other than the primary ones may also be considerably reduced; however, this may not be reflected in the phenotype. Many cases of primary gamma sarcoglycan deficiency showed normal or only mildly abnormal delta sarcoglycan staining. PMID- 10807698 TI - The principles of clinical cytogenetics PMID- 10807697 TI - Glycine to tryptophan substitution in type I collagen in a patient with OI type III: a unique collagen mutation. AB - We report a unique glycine substitution in type I collagen and highlight the clinical and biochemical consequences. The proband is a 9 year old Turkish boy with severely deforming osteogenesis imperfecta (OI). Biochemical analysis of (pro) collagen type I from a skin fibroblast culture showed both normal and overmodified alpha chains. Molecular analysis showed a G>T transversion in the COL1A2 gene, resulting in the substitution of glycine by tryptophan at position 277 of the alpha2(I) collagen chain. Glycine substitutions in type I collagen are the most frequent cause of the severe and lethal forms of OI. The phenotypic severity varies according to the nature and localisation of the mutation. Substitutions of glycine by tryptophan, which is the most voluminous amino acid, have not yet been identified in type I collagen or any other fibrillar collagen. The severe, though non-lethal OI phenotype associated with this mutation may appear surprising in view of the huge size of the tryptophan residue. The fact that the mutation resides within a so called "non-lethal" region of the alpha2(I) collagen chain supports a regional model in phenotypic severity for alpha2(I) collagen mutations, in which the phenotype is determined primarily by the nature of the collagen domain rather than the type of glycine substitution involved. PMID- 10807699 TI - Eye and face in syndromes - the clinical examination of eyes and their surroundings PMID- 10807700 TI - Detection of fetal cells in transcervical samples using X22 marker. PMID- 10807701 TI - Autosomal translocation associated with premature ovarian failure. PMID- 10807702 TI - Genotype-phenotype correlations in tuberous sclerosis. PMID- 10807703 TI - Where's the evidence? PMID- 10807704 TI - Periapical biopsy? PMID- 10807705 TI - Osseointegration is Latin. PMID- 10807706 TI - Periapical biopsy? PMID- 10807707 TI - Rapidly growing periodontal enlargement. PMID- 10807708 TI - An instrument for cross-cultural comparisons of the prevalence of temporomandibular disorder symptoms. PMID- 10807709 TI - Surgical risk factors for condylar resorption after orthognathic surgery. AB - OBJECTIVE: The purpose of this study was to look for surgical risk factors for condylar resorption after orthognathic surgery. STUDY DESIGN: Seventeen patients of a group of 452 patients who had undergone orthognathic surgery consecutively and who were in accordance with the inclusion criteria of this study showed postoperative condylar resorption (group I). Preoperative cephalometric characteristics and surgically induced movements of this group were compared with a control group of 17 of 452 patients (group II) in whom postoperative condylar resorption and skeletal relapse did not develop, despite mandibular retrognathism (ANB angle >4 degrees) and high mandibular plane angle (>40 degrees). RESULTS: The kind of osteosynthesis used was not significantly different between the 2 groups. The amount of surgical advancements and the vertical movements of the jaws were not significantly different between the 2 groups. However, the distal (P =.005) and proximal (P =.007) mandibular segments were rotated significantly further counterclockwise in group I. Surgically induced posterior condylar displacement occurred significantly more frequently (P =.007) in group I. CONCLUSIONS: Counterclockwise rotation of the distal and proximal mandibular segments and surgically induced posterior condylar displacement seem to be important surgical risk factors for postoperative condylar resorption. Therefore, these movements seem to be contraindicated in patients who are at high risk. PMID- 10807710 TI - Etiology and management of mandibular fractures associated with endosteal implants in the atrophic mandible. AB - Mandibular fractures can occur with the insertion of endosseous implants. Four patients whose mandibles were fractured with the removal or insertion of mandibular endosseous implants are described. Three of the patients required an autogenous bone graft to repair the fracture, and 1 patient was managed with a reconstruction plate. Strategies for prevention and treatment of this uncommon complication are discussed. PMID- 10807711 TI - Articulatory function in patients who have undergone glossectomy with use of an artificial graft membrane. AB - OBJECTIVE: The purpose of this study was to investigate, by assessing speech intelligibility, the long-term usefulness of an artificial bilayer membrane as a mucosal substitute after glossectomy. STUDY DESIGN: Artificial membranes were grafted in 8 patients with intraoral defects after glossectomy. Five patients served as a control group and received only standard closure by suture after glossectomy. Speech intelligibility was evaluated by means of a standardized Japanese speech intelligibility test 6 months or more after the operation. RESULTS: The postoperative courses of all 13 patients were uneventful. Intelligibility scores of those who received grafts were better than those of the control group in the overall score, the glossal sounds score, and the rear portion sounds score. CONCLUSION: The artificial membrane was useful for the glossectomy as a long-term mucosal substitute, as well as for the short-term. PMID- 10807712 TI - Insulin-dependent diabetes mellitus and oral soft tissue pathologies. I. Prevalence and characteristics of non-candidal lesions. AB - OBJECTIVE: A large epidemiologic study on oral health has found that several oral soft tissue lesions were more prevalent in subjects with insulin-dependent diabetes mellitus than in control subjects without diabetes. Our objective in part I of this article is to characterize those lesions not associated with Candida. STUDY DESIGN: This cross-sectional study determined the prevalence and characteristics of oral soft tissue diseases identified during a comprehensive oral evaluation of 405 adult subjects with diabetes and 268 control subjects without diabetes. RESULTS: Twenty specific oral soft tissue lesions were identified. Nearly twice as many subjects with diabetes as subjects without diabetes were found to have one or more oral soft tissue lesions (44.7% vs 25.0%; P <.0001). Subjects with diabetes also had significantly higher prevalence rates for 7 lesions, 3 of which were non-candidal: fissured tongue, irritation fibroma, and traumatic ulcers. (Four lesions generally associated with Candida infection median rhomboid glossitis, denture stomatitis, generalized atrophy of the tongue papillae, and angular cheilitis-will be described in part II of this article.) There were no differences found between the subjects with diabetes and the control subjects for lichen planus, gingival hyperplasia, or salivary gland disease. CONCLUSIONS: Oral soft tissue lesions were seen more frequently in subjects with insulin-dependent diabetes than in the control subjects. Characterization of 3 non-candidal lesions suggests that they are associated with trauma, delayed healing, or both. PMID- 10807714 TI - Oral manifestations of patients with Turner syndrome. AB - OBJECTIVE: The purpose of this study was to conduct a complete analysis of orofacial abnormalities of patients with Turner syndrome. STUDY DESIGN: Twenty nine patients with Turner syndrome and 30 without Turner syndrome (control group) were studied. The subjects were screened for cariological and periodontal alterations, orthodontic anomalies, tooth crown morphologic condition, alveolar arch dimensions, and temporomandibular joint function. RESULTS: Significantly lower mean values for decayed, missing, and filled teeth were found for patients with Turner syndrome as compared with the control group. Plaque and gingival indices were significantly higher in the study group. Orthodontic anomalies were more frequent and severe in patients with Turner syndrome. Our investigation showed reduced tooth crown size in the study group. The alveolar arch of the maxilla was narrower and of normal length, but the mandibular arch was shorter and broader. There was no significant alteration in temporomandibular joint function in the 2 examined groups. CONCLUSIONS: Our examination showed the importance of early diagnosis of oral anomalies and timely treatment of dental problems for patients with Turner syndrome. PMID- 10807713 TI - Insulin-dependent diabetes mellitus and oral soft tissue pathologies: II. Prevalence and characteristics of Candida and Candidal lesions. AB - OBJECTIVE: To assess the prevalence of Candida albicans and oral infection with Candida in patients with insulin-dependent diabetes mellitus (IDDM). STUDY DESIGN: This cross-sectional study compared the prevalence of candidiasis in 405 subjects with IDDM and 268 nondiabetic control subjects. Assessments included evidence of clinical manifestations of candidiasis and a quantitative measure of Candida pseudohyphae in a cytologic smear from the midline posterior dorsal tongue. RESULTS: More subjects with IDDM than control subjects without IDDM (15.1% vs 3.0%) were found to have clinical manifestations of candidiasis, including median rhomboid glossitis, denture stomatitis, and angular cheilitis. IDDM subjects were also more likely to have any Candida pseudohyphae in their cytologic smears (23.0% vs 5.7%; P <.0001), as well as pseudohyphae counts of >10/cm(2) (7.1% vs 0.8%; P <.0001). Diabetic subjects with median rhomboid glossitis were more likely to have a longer duration of IDDM and complications of nephropathy and retinopathy. Denture stomatitis was associated with smoking, retinopathy, higher Candida counts, poor glycemic control, and longer duration of IDDM. A multivariate regression analysis found 3 factors to be significantly associated with the presence of Candida pseudohyphae in the subjects with IDDM: current use of cigarettes (odds ratio, 2:4), use of dentures (odds ratio, 2:3), and elevated levels of glycosylated hemoglobin (odds ratio, 1:9). The use of antimicrobials, immunosuppressants, or drugs with xerostomic side effects was not related to the presence of Candida. CONCLUSIONS: Candida pseudohyphae and oral soft tissue manifestations of candidiasis were more prevalent in subjects with IDDM than in control subjects without diabetes. The presence of Candida pseudohyphae was significantly associated with cigarette smoking, use of dentures, and poor glycemic control. PMID- 10807715 TI - Lichen planus--specific antigen in oral lichen planus and oral lichenoid drug eruptions. AB - OBJECTIVE: To investigate the usefulness of lichen planus-specific antigen as a marker to distinguish idiopathic oral lichen planus from oral lichenoid drug eruptions. STUDY DESIGN: Biopsy samples were taken from 6 patients with oral lichenoid drug eruptions and 6 patients with idiopathic oral lichen planus. Each biopsy sample was examined for the presence of lichen planus-specific antigen by using a modification of a previously described immunofluorescence method that uses autologous serum and also allogenic sera from the remaining 11 cases. RESULTS: All autologous and allogenic immunofluorescence tests showed negative findings for lichen planus-specific antigen. CONCLUSIONS: Lichen planus-specific antigen is not a useful marker to distinguish oral lichenoid drug eruptions from idiopathic lichen planus. This finding is in contrast with our findings in an earlier study of basal cell cytoplasmic autoantibodies. PMID- 10807716 TI - The relation between salivary IgA and caries in renal transplant patients. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of immunosuppressive drugs on the level of salivary immunoglobulin A (IgA) in patients who have received kidney transplants and the relation between the levels of salivary IgA and dental caries incidence. STUDY DESIGN: Patients who had undergone renal transplantation (n = 28, aged 18-54) were divided into 3 groups according to postsurgical period (0-6 months [G(1)], 6-12 months [G(2)], and >12 months [G(3)]). A healthy control group (n = 10, aged 17-49) was also included in this study. Saliva samples were collected from all patients by the spitting method. After collection, the samples were frozen immediately at -40 degrees C until analysis by the single radial immunodiffusion method. All fissure caries were examined clinically, and proximal caries were examined clinically and radiographically; caries status was determined according to the decay surface index. The findings were evaluated statistically by means of correlation analysis, the Kolmogorov-Smirnov test, and the 1-way Kruskal Wallis analysis of variance method. RESULTS: Salivary IgA levels of the patients who had undergone renal transplantation were found to be significantly lower than those of the control patients (G(1) = 6.76 mg/dL, G(2) = 6.80 mg/dL, G(3) = 7.84 mg/dL, and control group = 10.84 mg/dL, P <.001). However, the caries status of the patients who had undergone renal transplantation was not different from that of the control subjects for the first year after the transplant operation. The salivary IgA values of the 3 groups of patients who had undergone transplantation were not significantly different from each other. Thus, it was observed that a decrease in the level of salivary IgA does not result in an increase in caries incidence within 12 months after renal transplantation. The caries rate in the third group of patients who had undergone renal transplantation was found to be significantly different from those in the first and second groups. CONCLUSION: Low salivary IgA levels caused by immunosuppression are not correlated or associated with higher levels of dental caries within the first 12 months after renal transplantation. However, the incidence of dental caries was higher for patients who had undergone renal transplantation than for control subjects 12 months after renal transplantation. Because of the diagnostic processes used, dental caries may not become evident until after 12 months. PMID- 10807717 TI - Melanotic neuroectodermal tumor of infancy in the mandible: report of a case. AB - A case of melanotic neuroectodermal tumor of infancy occurring in the mandible is described. The patient was a 1-month-old boy with a rapidly growing tumor of the mandible. Computed tomography showed 2 well-defined osteolytic lesions in the right mandible. Histopathologic diagnosis of a biopsy specimen was melanotic neuroectodermal tumor of infancy. The tumor was excised with removal of the surrounding bone, but 1(1/2) months later it recurred, and segmental mandibulectomy and reconstruction of the defect with a titanium miniplate was performed. Retrospectively, evidence of recurrence was noted on computed tomography taken on the tenth postoperative day. The recurrence was caused by incomplete removal of the tumor. Histopathologically, the tumor cells of the recurrent lesion were dispersed extensively in the bone marrow, and bone remodeling was active. The surgical procedure may have stimulated tumor cell proliferation and reactive bone formation. The patient was followed for 2 years with no evidence of recurrence or metastasis. PMID- 10807718 TI - Subacute necrotizing sialadenitis: report of 7 cases and a review of the literature. AB - Subacute necrotizing sialadenitis (SANS) is a nonspecific inflammatory condition of unknown etiology affecting oral minor salivary glands. The lesion is most often characterized by a localized palatal swelling, accompanied by an abrupt onset of pain. In this report, we describe the clinical, microscopic, and ultrastructural features of 7 new cases of SANS and combine them with those of 15 cases previously published in the English language medical literature. Patient ages ranged from 15 to 45 years, with a mean age of 21.9 years. A male to female ratio of 3.4 to 1 was observed, but probably reflects a male sampling bias because 14 of the 22 cases occurred in a military population. Seventeen cases occurred on the hard palate, 4 on the soft palate, and 1 on the tonsillar pillar. The lesions were nonulcerated swellings ranging in size from 0.3 to 2.5 cm in diameter. All lesions except one were painful, and most had been present for a week or less at the time of diagnosis. Histopathologic features included diffuse involvement of minor salivary glands by lymphocytes, histiocytes, neutrophils, and variably by eosinophils. Loss of acinar cells, early acinar cell necrosis, and atrophy of ductal cells were also observed. Although possible viral particles have been reported ultrastructurally in some cases of SANS, none were observed in our study. SANS appears to be a self-limiting process with most cases resolving 2 to 3 weeks after biopsy. SANS has been questioned as an entity by some investigators who prefer to include it within the spectrum of necrotizing sialometaplasia. However, we believe there is justification for the separation of these 2 entities based on clinical and histopathologic parameters. Although the specific etiology of SANS remains unknown, it most likely represents an infectious process or perhaps an immune response to an unknown allergen. PMID- 10807719 TI - Accessory parotid gland masses. AB - Three cases of accessory parotid gland lesions are reported. The literature concerning accessory gland disease and its diagnosis and treatment is reviewed. PMID- 10807720 TI - P53 and Ki-67 antigen expression in small oral biopsy specimens of salivary gland tumors. AB - OBJECTIVE: To investigate the possibility that various salivary gland tumors that look histologically similar could express p53 oncoprotein and Ki-67 proliferation antigen differentially and possibly aid in distinguishing benign from malignant lesions. STUDY DESIGN: Intraoral paraffin-embedded biopsy specimens of salivary gland tumors were used. Thirty-eight pleomorphic adenomas, 17 adenoid cystic carcinomas, 23 monomorphic adenomas, and 17 polymorphous low-grade adenocarcinomas were stained with p53 and Ki-67 antibodies by using an immunoperoxidase detection system. Each case was evaluated in terms of staining intensity and percentage of cells staining. RESULTS: Ki-67 and p53 antigens are expressed in generally low levels in the histologically well-differentiated salivary tumors that were studied here, both benign and malignant. Only 1 solid type adenoid cystic carcinoma showed a high percentage of cells expressing p53. CONCLUSIONS: The histologically well-differentiated salivary tumors studied do not show differential expression of p53 and Ki-67, in spite of their differing courses of biologic behavior. These antibodies should not be relied on to distinguish benign from malignant lesions of the salivary glands; however, they might be markers for those lesions that are dedifferentiating histologically and, therefore, might be displaying more aggressive behavior. PMID- 10807721 TI - Relationship of intracanal pressure with viscosity of endodontic sealer during warm Gutta-Percha vertical compaction. AB - OBJECTIVE: To test the association between endodontic sealer viscosity and intracanal pressure during warm gutta-percha vertical compaction. STUDY DESIGN: An artificial tooth model and pressure-sensing apparatus were developed so that reproducible, quantitative measurements could be determined. Intracanal pressure was measured at 3 phases of obturation: cone insertion, heat application, and force application. Samples were randomly assigned to 3 sealer viscosity groups (most viscous, normal, least viscous) and to a no-sealer group. Evaluations were blinded to group assignments (4 groups, 15 samples per group). RESULTS: Differences in intracanal pressure were detected between sealer viscosity groups. The most viscous group had the largest intracanal pressure. Throughout the phases of obturation, intracanal pressure did not remain constant; values at the time of heat application were lower than at cone insertion and force application. CONCLUSIONS: The results indicate that intracanal pressure increases as sealer viscosity is increased, and that intracanal pressure does not remain constant during obturation. PMID- 10807722 TI - Reaction of bony tissue to implanted silver glass ionomer and a reinforced zinc oxide-eugenol cement. AB - OBJECTIVE: To evaluate the tissue responses to implants of Ketac Silver and Super EBA cement in the guinea pig mandible. STUDY DESIGN: Sixteen guinea pigs were used for 2 experimental periods of 4 and 12 weeks. Both materials were placed in Teflon applicators and implanted into surgically prepared sites in the mandible. A histologic examination for reaction to the material occurred after the animals were killed and processed. RESULTS: After 4 weeks, minimal inflammatory reactions were observed in Ketac Silver implants, whereas the Super EBA implants showed minimal to moderate inflammation. Localized foreign body reaction was present in areas of fragmented small particles of Ketac Silver. At 12-weeks observation, no inflammatory reactions were present around either material. Direct bone contact was observed in 1 Ketac Silver implant. CONCLUSIONS: Ketac Silver and Super EBA cement elicited mild reactions under the conditions of this model. From a biologic point of view, these 2 materials may offer equal utility in endodontic surgery. PMID- 10807723 TI - Selection criteria for dental implant site imaging: a position paper of the American Academy of Oral and Maxillofacial radiology. AB - The American Academy of Oral and Maxillofacial Radiology (AAOMR) has reviewed the implant imaging literature and issues this position paper for preoperative assessment of the dental implant site. The history of dental implant radiology and basic imaging principles are reviewed. An overview is presented of anatomically salient features in the maxilla and mandible germane to implant imaging, specifically to cross-sectional or transverse imaging for endosseous implants. All current modalities, including intraoral, panoramic, cephalometric, tomographic, and computed tomography, are discussed in light of the imaging data needed to select optimum implant solutions. After reviewing the current literature, the AAOMR recommends that some form of cross-sectional imaging be used for implant cases and that conventional cross-sectional tomography be the method of choice for gaining this information for most patients receiving implants. PMID- 10807724 TI - New presidents of american academy of oral and maxillofacial radiology and american board of oral and maxillofacial radiology PMID- 10807725 TI - The sensitivity, specificity, and reliability of radiographic periapical diagnosis of posterior teeth. AB - PURPOSE: The purpose of this study was to define the percent accuracy, sensitivity, specificity, and reliability of periapical radiographic diagnosis of periapical inflammatory disease. METHOD: A total of 140 cases were reviewed under ideal viewing conditions by 6 observers. For each case, observers determined whether periapical disease was evident and rated the confidence of their interpretation. RESULTS: The mean percent accuracy was 70.2%. The mean sensitivity and specificity were 0.65 and 0.78, respectively. The intraclass correlation coefficients for intraobserver and interobserver reliability were 0.66 and 0.54, respectively. CONCLUSION: Specificity of periapical radiographic diagnosis is higher than sensitivity. These results support the current guideline for selective use of periapical views based on patient symptoms and clinical signs. Finally, dentists have a good degree of reliability on repeated evaluations, but only fair agreement when compared with other dentists reading the same radiograph. PMID- 10807726 TI - Central skull base invasion of maxillofacial tumors: computed tomography appearance. AB - OBJECTIVE: The aim of this study was to demonstrate computed tomography (CT) features of maxillofacial tumors invading the central skull base and to offer information in aiding diagnosis. METHODS: Fifty-eight maxillofacial tumors with central skull base invasion shown on axial and coronal CT images were divided into benign (13 cases) and malignant (45 cases) groups, based on their pathologic outcomes as proven by biopsy and surgery. RESULTS: Four manifestations of the skull base abnormality were observed on CT images: (1) resorption of central skull base (53 cases, 11 benign and 42 malignant tumors); (2) enlargement of the foramen and canal in the central skull base (20 cases, 4 benign and 16 malignant tumors); (3) thinning of the central skull base (11 cases, 8 benign and 3 malignant tumors); and (4) displacement of the central skull base (5 cases, all benign tumors). The following structures of the central skull base were involved: the roof of the pterygoid process of the sphenoid bone (49 cases), the sphenoid greater wing (37 cases), the sphenoid body and sinus (33 cases), the petrous apex (12 cases), the clivus (4 cases), and the articular surface of the temporal squama (3 cases). Of 58 patients, 30 (7 benign and 23 malignant) had tumors with central skull base erosion that infiltrated into the cranial cavity. CONCLUSION: Benign maxillofacial tumors with center skull base erosion tended to lead to the displacement and thinning of the base of the middle skull fossa in contrast with tumors with malignant maxillofacial tumors. It is believed that these CT manifestations might be valuable in making a differential diagnosis. PMID- 10807727 TI - Radiographic findings for solitary plasmacytoma of the bone in the anterior wall of the maxillary sinus: A case report. AB - Radiographic findings for a solitary plasmacytoma of the anterior wall of the maxillary sinus are reported. The diagnostic evaluation for this disease is discussed through use of plain images, computed tomography, and magnetic resonance imaging. The treatment selected was radiation therapy combined with chemotherapy. Computed tomography and magnetic resonance imaging revealed bone destruction, though this was not apparent on plain images. T(1)-weighted magnetic resonance images showed similar or high signal intensity relative to muscle; T(2) weighted images showed hyperintensity. PMID- 10807728 TI - Natural history and histological classification of atherosclerotic lesions: an update. PMID- 10807729 TI - Protein tyrosine phosphatases in the vessel wall: counterpoint to the tyrosine kinases. PMID- 10807730 TI - Hyperhomocysteinemia and oxidative stress: time for a reality check? PMID- 10807731 TI - 1999 George Lyman Duff memorial lecture: lipid transfer proteins, HDL metabolism, and atherogenesis. AB - Plasma high density lipoprotein (HDL) levels show an inverse relationship to atherogenesis, in part reflecting the role of HDL in mediating reverse cholesterol transport. The transfer of HDL cholesterol to the liver involves 3 catabolic pathways: the indirect, cholesteryl ester transfer protein (CETP) mediated pathway, the selective uptake (scavenger receptor BI) pathway, and a particulate HDL uptake pathway. The functions of the lipid transfer proteins (CETP and phospholipid transfer protein) in HDL metabolism have been elucidated by genetic approaches in humans and mice. Human CETP deficiency is associated with increased HDL levels but appears to increase coronary artery disease risk. Phospholipid transfer protein deficiency, produced by gene knockout in mice, results in decreased HDL levels, reflecting decreased transfer of phospholipids from triglyceride-rich lipoproteins into HDL. Obese (ob/ob) mice have markedly increased HDL levels and represent an interesting model of defective HDL catabolism in the liver. In hepatocytes of wild-type mice, there is extensive uptake and resecretion of HDL and selective uptake of cholesteryl ester from HDL during recycling. In ob/ob mice, these processes are defective, suggesting that HDL recycling plays an important role in holo-HDL catabolism, selective uptake, and the determination of plasma HDL levels. PMID- 10807732 TI - Protein-tyrosine phosphatases in the vessel wall: differential expression after acute arterial injury. AB - Many protein-tyrosine phosphatases (PTPases) have now been identified, but little is known about PTPase expression and regulation in vascular tissue and in vascular disease. Polymerase chain reaction (PCR) amplification and cDNA fingerprinting of PTPase catalytic domains, combined with random sequencing of PCR product libraries, identified 18 (8 receptor-like and 10 cytosolic) PTPases in the rat carotid artery and revealed differential expression of 5 of these PTPases during neointima formation after balloon catheter injury. In situ hybridization was used to localize mRNA expression in vessel cross sections for the 5 differentially expressed PTPases. This revealed that for 3 PTPases (SHP1, CD45, and PTPbeta), differential transcript abundance was due to appearance/loss of the cell types by which they were expressed (leukocytes for SHP1 and CD45, endothelial cells for PTPbeta). However, mRNA expression of 2 PTPases (PTPL1 and PTP1B) was specifically upregulated by proliferating and migrating smooth muscle cells (SMCs) in characteristic temporal and regional patterns in response to vessel damage. Quantitative PCR analysis showed that PTP1B and PTPL1 were induced approximately 30-fold and approximately 60-fold, respectively, by 2 weeks after injury in the damaged vessels compared with the uninjured vessels. PTP1B was rapidly upregulated in the media after vessel injury and remained highly expressed in the developing neointima. By contrast, PTPL1 expression did not increase dramatically until the SMCs had migrated into the intima. The differential expression of PTP1B and PTPL1 by SMCs after injury suggests roles for these PTPases in the regulation of vessel wall remodeling. PMID- 10807733 TI - Relationship between homocysteine and superoxide dismutase in homocystinuria: possible relevance to cardiovascular risk. AB - A modest homocysteine elevation is associated with an increased cardiovascular risk. Marked circulating homocysteine elevations occur in homocystinuria due to cystathionine beta-synthase (CbetaS) deficiency, a disorder associated with a greatly enhanced cardiovascular risk. Lowering homocysteine levels reduces this risk significantly. Because homocysteine-induced oxidative damage may contribute to vascular changes and extracellular superoxide dismutase (EC-SOD) is an important antioxidant in vascular tissue, we assessed EC-SOD and homocysteine in patients with homocystinuria. We measured circulating EC-SOD, total homocysteine (free plus bound), and methionine levels during the treatment of 21 patients with homocystinuria, 18 due to CbetaS deficiency, aged 8 to 59 years, and 3 with remethylating defects. We measured total homocysteine by immunoassay, EC-SOD by ELISA, and methionine by amino acid analysis and assessed interindividual and intraindividual relationships. There was a significant, positive relationship between EC-SOD and total homocysteine. For the interindividual assessment, levels were highly correlated, r=0.746, N=21, P<0.0001. This relationship was maintained after taking into account intraindividual patient variation (r=0.607, N=62, P<0.0001). In 2 newly diagnosed CbetaS-deficient patients, treatment that lowered the markedly elevated pretreatment homocysteine level (from 337 to 72 and from 298 to 50 micromol/L) reduced the associated elevated EC-SOD in each by 50%. EC SOD and methionine levels were unrelated (r=0.148, n=39, P=0.368). The positive relationship between circulating EC-SOD and homocysteine could represent a protective antioxidant response to homocysteine-induced oxidative damage and contribute to reducing cardiovascular risk in homocystinuric patients. EC-SOD levels may be relevant to the pathogenesis of vascular disease in other patient groups. PMID- 10807734 TI - Simultaneously increased TxA(2) activity in isolated arterioles and platelets of rats with hyperhomocysteinemia. AB - We aimed to elucidate the effect of hyperhomocysteinemia (HHcy) on the synthesis of prostaglandins in rat skeletal muscle arterioles and platelets. Male Wistar rats were divided into 2 groups: (1) control rats, with plasma Hcy levels of 6.5+/-0.5 micromol/L (n=50) and (2) rats with HHcy, induced by daily intake of 1 g/kg body weight methionine in the drinking water for 4 weeks (plasma Hcy levels were 20.6+/-3.0 micromol/L, P<0.01 versus controls; n=50). Arterioles (diameter approximately 130 micrometer) were isolated from the gracilis muscle, cannulated, and pressurized (at 80 mm Hg), and changes in their diameters were followed by video microscopy. Constrictions to bradykinin (BK; 10(-10) to 10(-7) mol/L) were significantly greater in HHcy than in control rat arterioles (at 10(-9) mol/L BK, changes were 11+/-3% in control and 41+/-9% in HHcy rats). The cyclooxygenase inhibitor indomethacin (10(-5) mol/L), the prostaglandin H(2)/thromboxane A(2) (PGH(2)/TxA(2)) receptor antagonist SQ 29,548 (10(-6) mol/L), or the TxA(2) synthase inhibitor furegrelate (5x10(-6) mol/L) significantly decreased constrictions to BK in both groups but more so in HHcy arterioles, thus eliminating the difference between responses of HHcy and control arterioles. Constrictions to U46619 (a TxA(2) analogue) were significantly greater in HHcy than in control arterioles (at 10(-8) mol/L U46619, values for controls were 33+/ 2% and 54+/-3% for HHcy). Endothelium removal or indomethacin treatment attenuated constrictions to U46619 in HHcy arterioles and eliminated the difference in responses. Also, aggregation of platelets from HHcy rats to collagen and ADP was significantly enhanced compared with controls (with 5 microgram/mL collagen: controls, 23+/-5%; HHcy, 49+/-5%; with 10(-7) mol/L ADP: controls, 25+/-3%; HHcy, 35+/-3%). Indomethacin or SQ 29,548 caused greater inhibition of aggregation of HHcy platelets compared with controls, thereby eliminating the differences between the 2 groups. Thus, HHcy enhances TxA(2) synthesis both in the arteriolar endothelium and platelets. By promoting vascular constriction and platelet aggregation simultaneously, these alterations are likely to contribute to the atherothrombotic vascular diseases described in HHcy. PMID- 10807735 TI - Heme oxygenase-1 is a cGMP-inducible endothelial protein and mediates the cytoprotective action of nitric oxide. AB - Inducible heme oxygenase (HO-1) has recently been recognized as an antioxidant and cytoprotective gene. By use of Western blotting, cell viability analysis, and antisense technique, the present study investigates the involvement of HO-1 in endothelial protection induced by the clinically used nitric oxide (NO) donor molsidomine (specifically, its active metabolite 3-morpholinosydnonimine [SIN-1]) and the second messenger cGMP. In bovine pulmonary artery endothelial cells, SIN 1 and S-nitroso-N-acetyl-D,L-penicillamine (SNAP) at 1 to 100 micromol/L induced the synthesis of HO-1 protein in a concentration-dependent fashion up to 3-fold over basal levels. HO-1 induction by SIN-1 was inhibited in the presence of the NO scavenger phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide and the soluble guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazole[4, 3-a]quinoxalin-1-one. 8-Bromo-cGMP (1 to 100 micromol/L) and dibutyryl cGMP (1 to 100 micromol/L) as well as the activator of particulate guanylyl cyclase atrial natriuretic peptide (1 to 100 nmol/L) produced increases in HO-1 protein similar to those produced by SIN-1. SIN-1 and 8-bromo-cGMP increased heme oxygenase activity (bilirubin formation). Cytoprotection by NO donors was abrogated in the presence of the heme oxygenase inhibitor tin protoporphyrin IX. Pretreatment of cells with a phosphorothioate-linked HO-1 antisense oligonucleotide prevented protection by SIN-1 or 8-bromo-cGMP against tumor necrosis factor-alpha cytotoxicity, whereas sense and scrambled HO-1 were without effect under these conditions. Our results show for the first time that HO-1 is a cGMP-sensitive endothelial gene and establish conclusively a causal relationship between HO-1 induction and endothelial protection by the NO/cGMP system. By targeting cytoprotective HO-1, NO donors may therefore be expected to induce antioxidant, antiatherogenic, and anti-inflammatory effects. PMID- 10807736 TI - Doxazosin inhibits retinoblastoma protein phosphorylation and G(1)-->S transition in human coronary smooth muscle cells. AB - Previous studies have demonstrated that the alpha(1)-adrenergic receptor antagonist doxazosin (Dox) inhibits multiple mitogenic signaling pathways in human vascular smooth muscle cells. This broad antiproliferative activity of Dox occurs through a novel mechanism unrelated to its blocking the alpha(1) adrenergic receptor. Flow cytometry demonstrated that Dox prevents mitogen induced G(1)-->S progression of human coronary artery smooth muscle cells (CASMCs) in a dose-dependent manner, with a maximal reduction of S-phase transition by 88+/-10.5% in 20 ng/mL platelet-derived growth factor and 1 micromol/L insulin (P+I)-stimulated cells (P<0.01 for 10 micromol/L Dox versus P+I alone) and 52+/-18.7% for 10% FBS-induced mitogenesis (P<0.05 for 10 micromol/L Dox versus 10% FBS alone). Inhibition of G(1) exit by Dox was accompanied by a significant blockade of retinoblastoma protein (Rb) phosphorylation. Hypophosphorylated Rb sequesters the E2F transcription factor, leading to G(1) arrest. Adenoviral overexpression of E2F-1 stimulated quiescent CASMCs to progress through G(1) and enter the S phase. E2F-mediated G(1) exit was not affected by Dox, suggesting that it targets events upstream from Rb hyperphosphorylation. Downregulation of the cyclin-dependent kinase inhibitory protein p27 is important for maximal activation of G(1) cyclin/cyclin-dependent kinase holoenzymes to overcome the cell cycle inhibitory activity of Rb. In Western blot analysis, p27 levels decreased after mitogenic stimulation (after P+I, 43+/-1.8% of quiescent cells [P<0.01 versus quiescent cells]; after 10% FBS, 55+/-7.7% of quiescent cells [P<0. 05 versus quiescent cells]), whereas the addition of Dox (10 micromol/L) markedly attenuated its downregulation (after P+I, 90+/-8.3% of quiescent cells [P<0.05 versus P+I alone]; after 10% FBS, 78+/ 8.3% of quiescent cells [P<0.05 versus 10% FBS alone]). Furthermore, Dox inhibited cyclin A expression, an E2F regulated gene that is essential for cell cycle progression into the S phase. The present study demonstrates that Dox inhibits CASMC proliferation by blocking cell cycle progression from the G(0)/G(1) phase to the S phase. This G(1)-->S blockade likely results from an inhibition of mitogen-induced Rb hyperphosphorylation through prevention of p27 downregulation. PMID- 10807737 TI - Intracellular Ca(2+) handling in vascular smooth muscle cells is affected by proliferation. AB - Despite intensive interest in the dedifferentiation process of vascular smooth muscle cells, very little data are available on intracellular Ca(2+) signaling. The present study was designed to investigate the evolution of the intracellular Ca(2+) pools when rat aortic smooth muscle cells (RASMCs) proliferate and to define the mechanisms involved in the functional alterations. RASMCs were cultured in different conditions, and [Ca(2+)](i) was measured by use of fura 2. Expression of the sarco(endo)plasmic reticulum Ca(2+) pumps (SERCA2a and SERCA2b), Ca(2+) channels, the ryanodine receptor (RyR), and the inositol trisphosphate receptor (IP3R) was studied by reverse transcription-polymerase chain reaction and immunofluorescence. Antibodies specific for myosin heavy chain isoforms were used as indicators of the differentiation state of the cell, whereas an anti-proliferating cell nuclear antigen antibody was a marker of proliferation. SERCA2a, SERCA2b, RyR3, and IP3R-1 mainly were present in the aorta in situ and in freshly isolated RASMCs. These cells used the 2 types of Ca(2+) channels to release Ca(2+) from a common thapsigargin-sensitive store. Proliferation of RASMCs, induced by serum or by platelet-derived growth factor BB, resulted in the disappearance of RyR and SERCA2a mRNAs and proteins and in the loss of the caffeine- and ryanodine-sensitive pool. The differentiated nonproliferative phenotype was maintained in low serum or in cells cultured at high density. In these conditions, RyR and SERCA2a were also present in RASMCs. Thus, expression of RyR and SERCA2a is repressed by cell proliferation, inducing loss of the corresponding Ca(2+) pool. In arterial smooth muscle, Ca(2+) release through RyRs is involved in vasodilation, and suppression of the ryanodine sensitive pool might thus alter the control of vascular tone. PMID- 10807738 TI - Localization of CD9, an enhancer protein for proheparin-binding epidermal growth factor-like growth factor, in human atherosclerotic plaques: possible involvement of juxtacrine growth mechanism on smooth muscle cell proliferation. AB - Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), a member of the EGF family, has a potent mitogenic activity for vascular smooth muscle cells (SMCs). We previously reported that HB-EGF is involved in atherogenesis of human aorta and coronary arteries. ProHB-EGF (the membrane anchored form of HB-EGF) has also been demonstrated to possess a mitogenic activity, which is approximately 30-fold increased when coexpressed with CD9 in mouse L cells. Thus, in the process of atherogenesis, CD9 may be involved in the proliferation of SMCs. We immunohistochemically investigated the localization of CD9 and proHB-EGF in the human aorta and coronary arteries. In normal aorta and coronary arteries, CD9 immunostaining was virtually negative, whereas proHB-EGF immunostaining was positive, especially in the arteries of babies. In contrast, in atherosclerotic lesions, some intimal SMCs were strongly positive for CD9 and proHB-EGF immunostaining. The juxtacrine growth activities of human aortic SMCs were inhibited in vitro by adding neutralization antibodies for CD9 or adding the specific inhibitor of HB-EGF. Besides, coexpressed CD9 and proHB-EGF cells markedly incorporated [(3)H]thymidine into the SMCs. CD9 is localized immunohistochemically in the SMCs of the atherosclerotic aorta and coronary arteries. CD9, when coexpressed with proHB-EGF, enhances proHB-EGF activities for SMC growth in a so-called juxtacrine manner in vitro and may be involved in atherogenesis. PMID- 10807739 TI - Endothelin-1 and smooth muscle cells: induction of jun amino-terminal kinase through an oxygen radical-sensitive mechanism. AB - Endothelin-1 (ET-1) has been proposed to contribute to atherogenesis and plaque rupture in coronary heart disease through activation of mitogen-activated protein kinases (MAPKs) in smooth muscle cells (SMCs). Reactive oxygen species (ROS) have been shown to be important signal transduction molecules in SMCs. Thus, the present study aimed to assess the role of ROS in ET-1-mediated activation of c Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) 1/2. Rat SMCs were exposed to ET-1 over time at concentrations from 10(-6) to 10( 10) mol/L, and MAPK activity was quantified. Activation of JNK and ERK was observed with a maximum stimulation at 10(-7) mol/L ET-1. JNK and ERK were activated by ET-1 binding to a single receptor (ET-1A) but differed in their downstream mechanisms: only JNK activation was sensitive to the radical scavenger N-acetylcysteine and diphenylene iodonium, an inhibitor of NADPH oxidase, indicating a role for ROS. The downstream MAPK effector and proinflammatory transcription factor, the activator protein-1 complex, was maximally activated 2 hours after the addition of ET-1. It was mainly composed of the JNK substrate c Jun, and activation was also dependent on ROS formation. We suggest that plaque activation by ET-1 can be mediated through ROS. It can be hypothesized that the clinical benefit of antioxidants in the treatment of atherogenesis may partially depend on neutralization of ET-1-mediated ROS production. PMID- 10807740 TI - Fibroblast growth factor-2 selectively stimulates angiogenesis of small vessels in arterial tree. AB - There is a critical need for quantifiable models of angiogenesis in vivo, and in general, differential effects of angiogenic regulators on vascular morphology have not been measured. Because the potent angiogenic stimulators fibroblast growth factor (FGF)-2 (basic FGF) and vascular endothelial growth factor (VEGF) are reported to stimulate angiogenesis through distinct signaling pathways, we hypothesized that FGF-2 stimulates vascular morphology differently than does VEGF and that stimulation of angiogenesis by FGF-2 is directly correlated to FGF receptor density. FGF-2 was applied at embryonic day 7 (E7), E8, or E9 to the quail chorioallantoic membrane (CAM); subsequent response of the arterial tree was measured by the fractal dimension (D(f)), a mathematical descriptor of complex spatial patterns, and by several generational branching parameters that included vessel length density (L(v)). After application of FGF-2 at E7, arterial density increased according to D(f) as a direct function of increasing FGF-2 concentration, and FGF-2 stimulated the growth of small vessels, but not of large vessels, according to L(v) and other branching parameters. For untreated control specimens at E7, L(v) of small vessels and D(f) were 11.1+/-1.6 cm(-1) and 1.38+/ 0.01, respectively; at E8, after treatment with 5 microgram FGF-2/CAM for 24 hours, L(v) of small vessels and D(f) increased respectively to 22.8+/-0.7 cm(-1) and 1. 49+/-0.02 compared with 16.3+/-0.9 cm(-1) and 1.43+/-0.02 for PBS-treated control specimens. Application of FGF-2 at E8 and E9 did not significantly increase arterial density. By immunohistochemistry, the expression of 4 high affinity tyrosine kinase FGF receptors was significantly expressed at E7, when CAM vasculature responded strongly to FGF-2 stimulation, but FGF receptor expression decreased throughout the CAM by E8, when vascular response to FGF-2 was negligible. In conclusion, the "fingerprint" vascular pattern elicited by FGF 2 was distinct from vascular patterns induced by other angiogenic regulators that included VEGF(165), transforming growth factor-beta1, and angiostatin. PMID- 10807741 TI - Assay for human matrix gla protein in serum: potential applications in the cardiovascular field. AB - Matrix Gla protein (MGP) is synthesized in a vitamin K-dependent way in smooth muscle cells of the healthy vessel wall, and its mRNA transcription is substantially upregulated in atherosclerotic lesions. Here we report the preparation of a monoclonal antibody against human MGP and its use in an enzyme linked immunosorbent assay. The intra-assay and interassay coefficients of variation in serum samples were 5.4% and 12.6%, respectively, and the lower detection limit was 8.5% of the normal serum value. Individual within-day variations were <11% and did not show a distinct circadian pattern. Day-to-day variations in fasting morning samples were <8%. In a first explorative survey, serum MGP concentrations were found to be significantly increased in patients with severe atherosclerosis, whereas these values were normal in those with low bone mass and osteoporosis. This finding is consistent with the high MGP mRNA expression observed in atherosclerotic vessels and plaques. More elaborate studies are required to assess the potential clinical utility of this newly developed assay. PMID- 10807742 TI - Lessons from sudden coronary death: a comprehensive morphological classification scheme for atherosclerotic lesions. PMID- 10807743 TI - Mildly oxidized LDL induces expression of group IIa secretory phospholipase A(2) in human monocyte-derived macrophages. AB - Phospholipase A(2)s (PLA(2)s) constitute a family of enzymes that hydrolyze fatty acids of membrane phospholipids, thus initiating the synthesis of proinflammatory mediators. Various PLA(2)s have been detected in human atherosclerotic arteries (advanced lesions); however, only the secretory group of PLA(2) has been shown to specifically hydrolyze low density lipoprotein (LDL)-associated phospholipids and, as such, may play a potential role in atherogenesis. In the present study, we investigated the expression pattern of group IIa, IV, and V PLA(2)s in human macrophages, which are the key cells involved in the onset and perpetuation of atherosclerosis. Immunohistochemical staining by double labeling showed that the secretory nonpancreatic PLA(2) (snpPLA(2)) is detectable in macrophages in the intima of early atherosclerotic lesions. Reverse transcription-polymerase chain reaction analysis of RNA extracted from human monocytes clearly showed that expression of group IV PLA(2) was enhanced during differentiation into macrophages, with an onset of induction at days 2 to 3 of differentiation. Group V snpPLA(2) was constitutively expressed on differentiation, whereas the detection of group IIa snpPLA(2) was dependent on both differentiation and subsequent stimulation of macrophages. Indeed, the transcription of group IIa snpPLA(2) in macrophages was induced by treatment with minimally modified or mildly oxidized LDL, whereas native, extensively oxidized, or acetylated LDL had no effect. To our knowledge, this is the first report describing induction of group IIa snpPLA(2) expression in human monocyte-derived macrophages. The mRNA levels of cytosolic PLA(2) group IV and snpPLA(2) group V remained unchanged on LDL treatment. Thus, our results show that the expression of distinct PLA(2) enzymes is regulated not only during differentiation of monocytes into macrophages but also on exposure of macrophages to distinct LDL species. Consequently, our results indicate a potential role for both cytosolic and secretory PLA(2) enzymes in inflammation and in macrophage functions related to atherosclerosis, with a specific role for group IIa snpPLA2 in LDL scavenging. PMID- 10807744 TI - Human ApoA-IV overexpression in transgenic mice induces cAMP-stimulated cholesterol efflux from J774 macrophages to whole serum. AB - The role of apolipoprotein A-IV (apoA-IV) in lipoprotein metabolism has not been established. The aim of the present study was to investigate the role of apoA-IV in reverse cholesterol transport by comparing cellular cholesterol efflux to serum or serum fractions from control mice and from mice transgenic for human apoA-IV (HuA-IVTg mice). When Fu5AH hepatoma cells were used, the cholesterol efflux to serum from either control or transgenic mice was similar. When control J774 macrophage cells were used, a comparison of efflux to serum or lipoprotein deficient serum (LPDS) failed to demonstrate any differences between control and transgenic mice. In contrast, when the J774 cells were pretreated with cAMP, there was a stimulation of efflux to whole serum or LPDS from HuA-IVTg mice. cAMP treatment had no effect on efflux to serum or LPDS from control mice. Pretreatment of the cells with cAMP did not enhance the efflux response to high density lipoprotein isolated from HuA-IVTg mouse serum. Our results suggest that apoA-IV, unassociated with high density lipoprotein particles, is responsible for enhanced cholesterol efflux. This study illustrates the role of lipid-free apolipoproteins in mediating cellular cholesterol efflux with use of a biological fluid and is potentially of physiological relevance, especially in apolipoprotein rich extravascular fluids. PMID- 10807745 TI - Induction of IG9 monocyte adhesion molecule expression in smooth muscle and endothelial cells after balloon arterial injury in cholesterol-fed rabbits. AB - The expression of monocyte-specific adhesion molecules and chemokines by cell types within the vessel wall plays an important role in foam cell accumulation during atherosclerotic plaque development. We previously identified IG9, a novel monocyte adhesion protein that is expressed on endothelial cells (ECs) overlying human and rabbit advanced atherosclerotic plaques. The present study was designed to determine the temporal and spatial expression of IG9 and the chemokine, monocyte chemoattractant protein-1 (MCP-1), after balloon injury with (double injury) or without (single injury) prior air desiccation EC injury in the femoral arteries of rabbits fed a high-cholesterol diet. By immunohistochemical analyses, intense reactivity with monoclonal antibodies to IG9 and MCP-1 was detected 24 hours after single injury in medial smooth muscle cells (SMCs) and in SMCs of adventitial microvessels. However, monocyte infiltration of the tunica media was minimal or not detected in these sections. IG9 and MCP-1 antibody reactivity in vessel sections 28 days after single injury and 24 hours, 7 days, and 28 days after double injury was localized to medial and neointimal SMCs, foam cells, and luminal ECs overlying the plaques. Uninjured rabbit (cholesterol or normal diet) vessel sections exhibited minimal IG9 and MCP-1 immunostaining. In vitro studies using human aortic SMCs demonstrated IG9 protein induction after 24 hours of treatment with platelet-derived growth factor-BB and interferon-gamma or epidermal growth factor. IG9 expression was further increased by pretreatment of SMCs with the proatherogenic lipid, minimally oxidized low density lipoprotein. After balloon injury (24 hours), IG9 is induced in vascular SMCs before the detectable accumulation of monocytes within the vessel wall. Thus, the expression of IG9 by SMCs as well as by ECs may be an important factor in the accumulation of foam cells in atherosclerotic plaque development after arterial injury. PMID- 10807746 TI - Well-defined regions of apolipoprotein B-100 undergo conformational change during its intravascular metabolism. AB - Apolipoprotein B (apoB)-100-containing lipoproteins are secreted from the liver as large triglyceride-rich very low density lipoproteins (VLDLs) into the circulation, where they are transformed, through the action of lipases and plasma lipid transfer proteins, into smaller, less buoyant, cholesteryl ester-rich low density lipoproteins (LDLs). As a consequence of this intravascular metabolism, apoB-containing lipoproteins are heterogeneous in size, in hydrated density, in surface charge, and in lipid and apolipoprotein composition. To identify specific regions of apoB that may undergo conformational changes during the intravascular transformation of VLDLs into LDLs, we have used a panel of 29 well-characterized anti-apoB monoclonal antibodies to determine whether individual apoB epitopes are differentially expressed in VLDL, intermediate density lipoprotein (IDL), and LDL subfractions isolated from 6 normolipidemic subjects. When analyzed in a solid phase radioimmunoassay, the expression of most epitopes was remarkably similar in VLDLs, IDLs, and LDLs. Two epitopes that are close to the apoB LDL receptor binding site show an increased expression in large (1.019 to 1.028 g/mL), medium (1.028 to 1.041 g/mL), and small (1.041 to 1.063 g/mL) LDLs compared with VLDLs and IDLs, and 2 epitopes situated between apoB residues 4342 and 4536 are significantly more immunoreactive in small and medium-sized LDLs compared with VLDLs, IDLs, and large LDLs. Therefore, as VLDL is converted to LDL, conformational changes identified by monoclonal antibodies occur at precise points in the metabolic cascade and are limited to well-defined regions of apoB structure. These conformational changes may correspond to alterations in apoB functional activities. PMID- 10807747 TI - In vivo metabolism of ApoB, ApoA-I, and VLDL triglycerides in a form of hypobetalipoproteinemia not linked to the ApoB gene. AB - Familial hypobetalipoproteinemia (FHBL) is an autosomal codominant disorder that may result from different mutations in the apolipoprotein B (apoB) gene or chromosome 2. However, linkage of FHBL to the apoB gene was ruled out in 2 kindreds reported to date, and the genetic and metabolic bases for FHBL remain unknown. One of the reported kindreds is our 40-member F kindred, in which we found linkage of FHBL to a novel susceptibility region on chromosome 3p21. 1-2. In addition to having low apoB levels, some, but not all, of the affected subjects in the F kindred also had low levels of high density lipoprotein (HDL) cholesterol and apoA-I. Our aim was to define the metabolic bases of the disorder in the F kindred. Therefore, we studied the in vivo kinetics of apoB and apoA-I and very low density lipoprotein (VLDL) triglycerides in 4 affected subjects and 5 normolipidemic relatives. Deuterated leucine and deuterated glycerol were used to label the apolipoproteins and triglycerides, respectively. Compartmental modeling was used to obtain the kinetic parameters. Affected subjects had (1) normal fractional catabolic rates (FCRs) for VLDL apoB, (2) increased FCRs for low density lipoprotein (LDL) apoB (0.050+/-0.009 versus 0. 030+/-0.006 pools per hour for normal subjects, P=0.005), and (3) decreased production rates of VLDL apoB (11.4+/-1.7 versus 25.6+/-4. 9 mg. kg(-1). d(-1), P=0.003), LDL apoB (7.8+/ 1.3 versus 12.7+/-3.7 mg. kg(-1). d(-1), P=0.04), and VLDL triglycerides (8.2+/ 4.5 versus 19.6+/-10.8 58 micromol. kg(-1). h(-1), P=0.09). These data differ from those obtained in previously studied FHBL heterozygotes bearing apoB-2 and apoB-9, 2 very short truncations of apoB. Low HDL cholesterol and apoA-I levels were caused by higher apoA-I FCRs (0. 035+/-0.005 versus 0.018+/-0.005 pools per hour in controls, P<0.01) without significant decrease in apoA-I production rates (18.7+/-2.7 versus 22.8+/-5.6 mg. kg(-1). d(-1)). In conclusion, decreased secretion of apoB-containing lipoproteins and hypercatabolism of LDL account for low apoB and cholesterol levels in this novel form of FHBL. PMID- 10807748 TI - Common polymorphism in promoter of microsomal triglyceride transfer protein gene influences cholesterol, ApoB, and triglyceride levels in young african american men: results from the coronary artery risk development in young adults (CARDIA) study. AB - The microsomal triglyceride transfer protein (MTP) plays a key role in the assembly of apolipoprotein B (apoB)-containing lipoproteins. We investigated the relation between lipid profiles and a common functional polymorphism (-493G/T) of the MTP gene in a large sample of young black men in the Coronary Artery Risk Development in Young Adults (CARDIA) Study. We performed serial cross-sectional analyses on lipids of 586 black men in 5 exams over 10 years of follow-up. Total cholesterol, LDL cholesterol, and apoB levels were very similar between the GT and GG genotypes; therefore, the GT and GG genotypes were combined as 1 group when the 3 phenotypes were analyzed. The results from ANCOVA showed that the TT group (prevalence 7%) had higher levels of apoB-related lipids than did the GT+GG group: the difference in total cholesterol ranged from 2 (P=0.79) to 19 (P=0.002) mg/dL in exams 1 to 5; the difference in LDL cholesterol ranged from 10 (P=0.14) to 17 (P=0.003) mg/dL in exams 1 to 4, but in examination 5, the difference became negligible. The TT group had higher levels of apoB, measured in only 2 exams, by 6 (P=0.12) and 9 (P=0.03) mg/dL. The TT group had higher levels of triglycerides than did the TG or GG group by 3 to 34 (P=0.02 to approximately 0.003) mg/dL in all 5 exams. HDL cholesterol and apolipoprotein A-I levels were similar among the 3 genotypes. Our serial cross-sectional analyses indicated that the TT genotype was associated with higher levels of total cholesterol, LDL cholesterol, triglycerides, and apoB in young black men. The broad effect of this polymorphism on several atherogenic traits suggests that the MTP gene could be influential in atherosclerosis. PMID- 10807749 TI - Association of cholesteryl ester transfer protein-TaqIB polymorphism with variations in lipoprotein subclasses and coronary heart disease risk: the Framingham study. AB - Cholesteryl ester transfer protein (CETP) facilitates the exchange of triglycerides and cholesteryl esters between lipoprotein particles, a key step in reverse cholesterol transport in humans. Variations at the CETP locus have been shown to be determinants of the levels and activity of CETP and high density lipoprotein (HDL) plasma concentration. The associations of the common CETP polymorphism, TaqIB in intron 1, with lipoprotein levels and particle size distribution, CETP activity, and coronary heart disease (CHD) risk were examined in a population-based sample of 1411 men and 1505 women from the Framingham Offspring Study. The B2 allele frequency was 0.444 in men and 0.433 in women, and its presence was significantly (P<0.05) associated with decreased CETP activity. B1B1 men had lower HDL cholesterol (HDL-C) levels (1.07 mmol/L) compared with B1B2 (1.14 mmol/L) and B2B2 (1.18 mmol/L) men (P<0.001). Likewise, B1B1 women had lower HDL-C levels (1.40 mmol/L) compared with B1B2 (1.46 mmol/L) and B2B2 (1.53 mmol/L) women (P<0.001). In men, the B2 allele was associated with increased particle size for HDL and low density lipoprotein. In women, a similar effect was demonstrated only for HDL particle size. The odds ratio for prevalent CHD associated with the B2 allele was 0.696 (P=0.035) in men. After adjusting for age, body mass index, systolic blood pressure, diabetes, smoking, alcohol consumption, beta-blocker use, total cholesterol, and HDL-C, this odds ratio was 0.735 (P=0.187), suggesting that the protective effect of the B2 allele was due in part to its association with HDL-C levels. No significant protective effects were observed in women. These data demonstrate that variation at the CETP gene locus is a significant determinant of HDL-C levels, CETP activity, and lipoprotein size in this population. Moreover, these effects appear to translate into a lower CHD risk among those men with the B2 allele. PMID- 10807751 TI - Functional characterization of 4 polymorphisms in promoter region of hepatic lipase gene. AB - Hepatic lipase (HL) is a lipolytic enzyme involved in the metabolism of plasma lipoproteins, especially high density lipoproteins. Association studies have provided strong evidence for relations of common mutations in the promoter region of the HL gene to postheparin plasma HL activity and the plasma high density lipoprotein cholesterol concentration, but the functional relevance of these polymorphisms has not been evaluated to date. We analyzed the physiological significance of 4 common polymorphisms (-250G/A, -514C/T, -710T/C, and -763A/G, all in strong linkage disequilibrium) in the promoter of the HL gene by use of electrophoretic mobility shift assays and transient transfection studies in HepG2 cells. No consistent evidence was found for a significant contribution of any of these polymorphisms to the basal rate of transcription of the HL gene. These data suggest that the 4 polymorphisms in the promoter region of the HL gene are in linkage disequilibrium with >/=1 as-yet-unknown functional polymorphisms in the HL gene locus with a significant effect on HL metabolism and/or enzymatic activity. PMID- 10807750 TI - Intronic polymorphism in the fatty acid transport protein 1 gene is associated with increased plasma triglyceride levels in a French population. AB - Fatty acids play important biological roles in cells. The precise mechanism whereby fatty acids cross the plasma membrane is still poorly understood. They can cross membranes because of their hydrophobic properties and/or be transported by specific proteins. Recently, a gene coding for fatty acid transport protein 1 (FATP1), an integral plasma membrane protein implicated in this process, was cloned in humans. We screened the gene by single-strand conformation polymorphism analysis and detected an A/G polymorphism in intron 8. We analyzed the potential relations of this genetic polymorphism with various obesity markers and with plasma lipid profiles in a random sample of 1144 French subjects aged 35 to 64 years. We detected statistically significant associations between this FATP1 A/G polymorphism and an increase in plasma triglyceride levels, mainly in women. These results suggest that genetic variability in the FATP1 gene may affect lipid metabolism, especially in women, and reinforce the potential implication of FATP1 in lipid homeostasis. PMID- 10807752 TI - Noncholesterol sterols and cholesterol lowering by long-term simvastatin treatment in coronary patients: relation to basal serum cholestanol. AB - Coronary patients with low baseline ratios of serum cholestanol and plant sterols to cholesterol (indicating low cholesterol absorption) but not those with high ratios (high absorption) experienced reduced recurrences of coronary events during simvastatin treatment in the Scandinavian Simvastatin Survival Study. Thus, in the present study, serum cholesterol, its precursor sterols (reflecting cholesterol synthesis), plant sterols (campesterol and sitosterol), and cholestanol were measured before and during a 5-year period of placebo treatment (n=433) and simvastatin treatment (n=434) in patients from a subgroup of the Scandinavian Simvastatin Survival Study to determine whether changes in cholesterol synthesis and serum levels were related to cholesterol absorption. Serum cholesterol level was unchanged, the ratios of cholesterol precursor sterols to cholesterol were decreased, and the ratios of plant sterols to cholesterol were increased in relation to increasing baseline ratios of cholestanol quartiles. The latter predicted 5-year ratios and simvastatin-induced reductions of the precursor sterols, with the lowering of the ratios (cholesterol synthesis reduction) being almost twice higher in the lowest versus the highest quartile. The ratios of plant sterols, especially campesterol, to cholesterol were markedly increased during simvastatin treatment, mostly in subjects with the highest baseline cholestanol quartiles. Simvastatin reduced serum cholesterol more (P=0.003) in the lowest versus the highest cholestanol quartile during the 5 year treatment period. The results show for the first time that baseline cholesterol metabolism, measured by serum noncholesterol sterols, predicts the effectiveness of simvastatin in reducing cholesterol synthesis and serum levels of cholesterol. The drug suppresses the synthesis of cholesterol markedly more effectively in subjects with high than with low baseline synthesis but reduces respective serum cholesterol levels less markedly than synthesis. Subjects with high cholesterol absorption and low synthesis may need a combination therapy to lower more effectively their serum cholesterol levels and prevent an increase in the levels of plant sterols. PMID- 10807753 TI - Antithrombotic effect of platelet glycoprotein Ib-blocking monoclonal antibody Fab fragments in nonhuman primates. AB - Platelet adhesion in arterial blood flow is mainly supported by the platelet receptor glycoprotein (GP) Ib, which interacts with von Willebrand factor (vWF) that is bound to collagen at the site of vessel wall injury. Antibody 6B4 is a monoclonal antibody (MoAb) raised against purified human GPIb. MoAb 6B4 inhibits both ristocetin- and botrocetin-induced, vWF-dependent human platelet agglutination. MoAb 6B4 furthermore blocks shear-induced adhesion of human platelets to collagen I. We studied the antithrombotic effect of this inhibitory murine MoAb 6B4 in a baboon model of arterial thrombosis. When injected into baboons, intact IgG and its F(ab')(2) fragments caused almost immediate thrombocytopenia, whereas injection of the Fab fragments alone did not. Fab fragments were subsequently used to investigate their in vivo effect on platelet deposition onto a thrombogenic device, consisting of collagen-rich, glutaraldehyde-fixed bovine pericardium (0.6 cm(2)), at a wall shear rate ranging from 700 to 1000 s(-1). Baboons were either pretreated with Fabs to study the effect of inhibition on platelet adhesion or treated 6 minutes after placement of the thrombogenic device to investigate the effect on interplatelet cohesion. Pretreatment of the animals with bolus doses ranging from 80 to 640 microgram/kg Fab fragments significantly reduced (111)In-labeled platelet deposition onto the collagen surface by approximately 43% to 65%. Only the highest dose caused a significant prolongation (doubling) of the bleeding time. Ex vivo ristocetin induced platelet agglutination was equally reduced. Treatment with a bolus of 110 microgram/kg Fab fragments after a thrombus was allowed to form for 6 minutes had no effect on further platelet deposition. We therefore conclude that Fab fragments or derivatives of inhibitory anti-GPIb antibodies may be useful compounds to prevent thrombosis. PMID- 10807754 TI - Influence of fibrin network conformation and fibrin fiber diameter on fibrinolysis speed: dynamic and structural approaches by confocal microscopy. AB - Abnormal fibrin architecture is thought to be a determinant factor of hypofibrinolysis. However, because of the lack of structural knowledge of the process of fibrin digestion, relationships between fibrin architecture and hypofibrinolysis remain controversial. To elucidate further structural and dynamic changes occurring during fibrinolysis, cross-linked plasma fibrin was labeled with colloidal gold particles, and fibrinolysis was followed by confocal microscopy. Morphological changes were characterized at fibrin network and fiber levels. The observation of a progressive disaggregation of the fibrin fibers emphasizes that fibrinolysis proceeds by transverse cutting rather than by progressive cleavage uniformly around the fiber. Plasma fibrin clots with a tight fibrin conformation made of thin fibers were dissolved at a slower rate than those with a loose fibrin conformation made of thicker (coarse) fibers, although the overall fibrin content remained constant. Unexpectedly, thin fibers were cleaved at a faster rate than thick ones. A dynamic study of FITC-recombinant tissue plasminogen activator distribution within the fibrin matrix during the course of fibrinolysis showed that the binding front was broader in coarse fibrin clots and moved more rapidly than that of fine plasma fibrin clots. These dynamic and structural approaches to fibrin digestion at the network and the fiber levels reveal aspects of the physical process of clot lysis. Furthermore, these results provide a clear explanation for the hypofibrinolysis related to a defective fibrin architecture as described in venous thromboembolism and in premature coronary artery disease. PMID- 10807755 TI - Expression, localization, and activity of tissue factor pathway inhibitor in normal and atherosclerotic human vessels. AB - Tissue factor (TF) pathway inhibitor (TFPI) is the major downregulator of the procoagulant activity of the TF-factor VIIa (FVIIa) complex (TF. FVII). The active TF present in the atherosclerotic vessel wall is proposed to be responsible for the major complication of primary atherosclerosis, namely, acute thrombosis after plaque rupture, but our knowledge of the sites of TFPI expression in relation to TF remains fragmentary. The aim of this study was to investigate the expression, localization, and activity of TFPI and its relation to the activity and distribution of TF in the normal and atherosclerotic vessel wall. We applied a novel approach in which serial cross sections of human vascular segments were used to perform a complete set of assays: immunolabeling for TFPI and/or TF, in situ hybridization for the expression of TFPI mRNA, ELISA for the determination of TFPI antigen, and functional assay for the activity of TFPI and TF. In healthy vessels, TFPI protein and mRNA are present in luminal and microvascular endothelial cells (ECs) and in the medial smooth muscle cells (SMCs). In atherosclerotic vessels, TFPI protein and mRNA frequently colocalized with TF in ECs overlying the plaque and in microvessels, as well as in the medial and neointimal SMCs, and in macrophages and T cells in areas surrounding the necrotic core. At the ultrastructural level, immunogold electron microscopy confirmed the localization of TFPI in ECs, macrophages/foam cells, and SMCs. In ECs and SMCs, the gold particles decorated the plasmalemma proper and the caveolae. ELISA on cross sections revealed that atherosclerotic tissues contain more TFPI than do the healthy vessels. TFPI was functionally active against TF. FVIIa-induced coagulation, and its activity was higher in those tissues that display less TF. The largest amount of TFPI and TF were detected in complicated arterial plaques. By immunofluorescence, TFPI colocalized with platelet- and fibrin-rich areas within the organized thrombi. Atherosclerotic vessel sections promote activation of factor X, which is dependent on the presence of TF and enhanced by preincubation of the sections with anti-TFPI IgG. Taken altogether, our results suggest that TFPI is largely expressed in the normal vessel wall and enhanced in the atherosclerotic vessel, in a manner suggesting a significant role of TFPI in the regulation of TF activity. PMID- 10807756 TI - Vascular endothelial growth factor production by fibroblasts in response to factor VIIa binding to tissue factor involves thrombin and factor Xa. AB - Tissue factor (TF) assembled with activated factor VII (FVIIa) initiates the coagulation cascade. We recently showed that TF was essential for FVIIa-induced vascular endothelial growth factor (VEGF) production by human fibroblasts. We investigated whether this production resulted from TF activation by its binding to FVIIa or from the production of clotting factors activated downstream. Incubation of fibroblasts with a plasma-derived FVIIa concentrate induced the generation of activated factor X (FXa) and thrombin and the secretion of VEGF, which was inhibited by hirudin and FXa inhibitors. By contrast, the addition of recombinant FVIIa to fibroblasts did not induce VEGF secretion unless factor X was present. Moreover, thrombin and FXa induced VEGF secretion and VEGF mRNA accumulation, which were blocked by hirudin and FXa inhibitors, respectively. The effect of thrombin was mediated by its specific receptor, protease-activated receptor-1; in contrast, the effect of FXa did not appear to involve effector cell protease receptor-1, because it was not affected by an anti-effector cell protease receptor-1 antibody. An increase in intracellular calcium with the calcium ionophore A23187 or intracellular calcium chelation by BAPTA-AM had no effect on either basal or FXa-induced VEGF secretion, suggesting that the calcium signaling pathway was not sufficient to induce VEGF secretion. Finally, FVIIa, by itself, had no effect on mitogen-activated protein (MAP) kinase activation, contrary to thrombin and FXa, which activate the p44/p42 MAP kinase pathway, as shown by the blocking effect of PD 98059 and by Western blotting of activated MAP kinases. These findings indicate that FVIIa protease induction of VEGF expression is mediated by thrombin and FXa generated in response to FVIIa binding to TF expressing fibroblasts; they also exclude a direct signaling involving MAP kinase activation via the intracellular domain of TF when expressed by these cells. PMID- 10807757 TI - Factor V antigen levels and venous thrombosis: risk profile, interaction with factor V leiden, and relation with factor VIII antigen levels. AB - Clotting factor V has a dual function in coagulation: after activation, procoagulant factor V stimulates the formation of thrombin, whereas anticoagulant factor V acts as a cofactor for activated protein C (APC) in the degradation of factor VIII/VIIIa, thereby reducing thrombin formation. In the present study, we evaluated whether plasma factor V levels, either decreased or increased, are associated with venous thrombosis. High procoagulant factor V levels may enhance prothrombinase activity and increase the thrombosis risk. Low anticoagulant factor V levels could reduce APC-cofactor activity in the factor VIII inactivation (APC-resistant phenotype), which might also promote thrombosis. Low factor V levels in combination with factor V Leiden could lead to a more severe APC-resistant phenotype (pseudohomozygous APC resistance). To address these issues, we have measured factor V antigen (factor V:Ag) levels in 474 patients with thrombosis and 474 control subjects that were part of the Leiden Thrombophilia Study (LETS). Factor V:Ag levels increased by 7.6 U/dL for every successive 10 years of age. Mean factor V:Ag levels were 134 (range 41 to 305) U/dL in patients and 132 (range 47 to 302) U/dL in controls. Neither high nor low factor V:Ag levels were associated with venous thrombosis. We found that factor V:Ag and factor VIII antigen levels in plasma were correlated, but factor V did not modify the thrombotic risk of high factor VIII levels. The normalized APC ratio was not influenced by the factor V:Ag level in subjects with or without factor V Leiden. We conclude that neither low nor high factor V:Ag levels are associated with venous thrombosis and that factor V:Ag levels do not mediate the thrombotic risk associated with high factor VIII levels. PMID- 10807758 TI - The A -844G polymorphism in the PAI-1 gene is associated with a higher risk of venous thrombosis in factor V Leiden carriers. AB - Identification of combined genetic factors in factor V Leiden carriers is important for a more accurate risk assessment for venous thrombosis (VT). Among these individuals, we evaluated the role of polymorphisms of the plasminogen activator inhibitor-1 (PAI-1) gene in the thrombophilic phenotype. A total of 382 factor V Leiden carriers were included in the study. This population was divided into 3 groups. Group 1 (n=168) included individuals with a personal history of VT; group 2 (n=140) included individuals without personal VT but with a familial history of VT; and group 3 (n=74) included individuals without VT and with a fortuitous discovery of the factor V Leiden mutation. We compared the genotype distribution of 2 polymorphisms, A -844G and -675 4G/5G, located in the promoter region of the PAI-1 gene among these 3 groups of individuals. The A -844G allele frequency differed significantly among the 3 groups (P=0.048), the A allele being more frequent in patients who suffered from VT (61%) than in subjects without VT (52%, P=0.015), whereas no difference was observed between the 2 groups of asymptomatic individuals. The prevalence of genotype AA carriers was higher in patients with VT (38%) than in asymptomatic individuals (21%, P=0. 015), leading to an odds ratio of 1.74 (95% confidence interval, 1.3 to 3.8). Carrying the AA genotype conferred a risk of deep VT of 2. 08 (95% confidence interval, 1.28 to 3.40), whereas it did not seem to significantly influence the risk of pulmonary embolism. Concerning the -675 4G/5G polymorphism, no significant difference was observed among the 3 groups, the 4G allele frequency being 0.54 (in group 1), 0.49 (in group 2), and 0.45 (in group 3). These data suggest a role for the A844G PAI-1 gene polymorphism in the thrombophilic phenotype of factor V Leiden carriers. PMID- 10807759 TI - Familial thrombophilia associated with homozygosity for the cystathionine beta synthase 833T-->C mutation. AB - Severe hyperhomocysteinemia due to cystathionine beta-synthase (CBS) deficiency is a strong risk factor for premature cardiovascular disease. Among untreated patients, approximately 50% have suffered a thromboembolic event by 30 years of age. We report on 3 sisters with severe hyperhomocysteinemia due to homozygosity for the CBS 833T-->C mutation. These patients, who displayed no other known thrombophilic predisposition, had suffered single or multiple venous thrombosis before CBS deficiency was diagnosed relatively late in life. In this family, homozygosity for the 833T-->C mutation was associated with a mild phenotype with respect to other sequelae of CBS deficiency. Consequently, our results indicate that most cases with this genotype may remain undiagnosed. Investigated family members heterozygous for the 833T-->C mutation displayed normal total homocysteine in plasma (tHcy) levels, even when they were homozygous for the methylenetetrahydrofolate reductase 677C-->T polymorphism. The prevalence of homozygosity for the 833T-->C mutation has previously been estimated at no less than 1:20 500 in our population. Because a reduction of the severely elevated levels of tHcy in CBS deficiency reduces cardiovascular risk and because homozygosity for the 833T-->C mutation is more prevalent than previously thought, our results emphasize the importance of measuring tHcy routinely in thrombophilia screening. PMID- 10807760 TI - Oral, but not transdermal, administration of estrogens lowers tissue-type plasminogen activator levels in humans without affecting endothelial synthesis. AB - Oral estrogen administration decreases plasma levels of tissue-type plasminogen activator (tPA), which may be explained by a decrease in endothelial tPA synthesis, an increase in its hepatic clearance, or both. In the present study, we determined (1) differences between oral (ie, via the liver) ethinyl estradiol and transdermal (ie, systemic) 17beta-estradiol administration on plasma antigen levels of tPA and plasminogen activator inhibitor type-1 before and after 4 months of hormone administration and (2) effects on endothelial tPA synthesis, by measuring the local increase in plasma tPA during venous occlusion of the upper extremity. Thirty transsexual males (median age 32 years, range 20 to 44 years ) were randomly assigned to either oral ethinyl estradiol (n=15) or transdermal 17beta-estradiol (n=15); both treatments included the antiandrogen cyproterone acetate (CA). Ten males were treated with CA alone. Seventeen transsexual females (median age 27 years, range 18 to 37 years) were treated with intramuscular testosterone esters. Only oral ethinyl estradiol plus CA but neither transdermal 17beta-estradiol plus CA, nor oral CA, nor parenteral testosterone lowered plasma tPA and plasminogen activator inhibitor-1 (P<0.001 for both). tPA release during venous occlusion was not affected by oral ethinyl estradiol plus CA in males (P=0.52) or by parenteral testosterone in females (P=0.89). These data are consistent with a previous observation, in rodents, that the decrease in tPA after oral estrogen administration can be explained by an increase in hepatic tPA clearance, leaving endothelial tPA synthesis unchanged, and suggest that these mechanisms also explain the decrease in tPA in humans. PMID- 10807761 TI - Postmenopausal hormone replacement therapy increases coagulation activity and fibrinolysis. AB - Hormone replacement therapy (HRT) appears to be cardioprotective in postmenopausal women; however, concerns exist over its thrombogenic effects. To address the effects of combined HRT on coagulation and fibrinolysis, we have measured circulating markers of these processes in a double-blind placebo controlled trial. Forty-two healthy postmenopausal women aged 50 to 75 years received continuous combined HRT with 2 mg estradiol+1 mg norethisterone or placebo for 6 weeks. Hormone profiles were measured at baseline, and lipid and hemostatic parameters were measured at baseline and after 6 weeks of therapy. Baseline characteristics were similar in the 2 groups. With change from baseline the main outcome measure, HRT increased the markers of coagulation (prothrombin fragments 1+2, 0.20+/-0.06 versus 0.06+/-0.04 nmol/L, P=0.0005; soluble fibrin, 2.3+/-0.4 versus 0.25+/-0.3 microgram/mL, P=0.0004), reduced plasma fibrinolytic inhibitory activity (plasminogen activator inhibitor-1, -0.67+/-0.16 versus 0.24+/-0.21 U/mL, P=0.002), and increased fibrinolysis (D-dimer, 24+/-12 versus 6+/-8 ng/mL, P=0.04) compared with placebo. Increases in soluble fibrin and D dimer were positively correlated (r=0.59, P=0.02), but changes in plasminogen activator inhibitor-1 and D-dimer were unrelated. Although baseline hemostatic and lipid parameters were correlated, there were no associations between changes in hemostatic markers and lipids after treatment. Short-term oral combined continuous HRT (estradiol and norethisterone) increased thrombin and fibrin generation, reduced plasma fibrinolytic inhibitory activity, and increased fibrinolysis. Enhanced fibrinolysis was related to increased fibrin generation but not reduced plasma fibrinolytic inhibitory activity. Coagulation activation may partly explain the increases in venous thrombosis and cardiovascular events reported with the use of combined HRT. PMID- 10807762 TI - Modulation of bcl-2 and p27 in human primitive proliferating hematopoietic progenitors by autocrine TGF-beta1 is a cell cycle-independent effect and influences their hematopoietic potential. AB - Primitive, proliferating hematopoietic progenitors (defined as cytokine low responding primitive progenitors; CLRPP), isolated from human CD34+ cells, expressed endoglin (CD105) and produced transforming growth factor-beta1 (TGF beta1). Culture of CLRPP in serum-free conditions with anti-TGF-beta1 monoclonal antibody produced a substantial decrease in bcl-2 protein/RNA levels and a significant reduction of cloning and long-term culture-initiating cell (LTC-IC) activities. GATA-1 and PU.1 RNA levels were significantly up-regulated in anti TGF-beta1-treated CLRPP, which generated an increased number of cells expressing CD15/CD11b/glycophorin-A. The described effects of TGF-beta1 neutralization were observed in the absence of any relevant effect on cell cycle; number of cell divisions; p53, c-myc, and p21 RNA levels; bcl-xL and bax protein levels; and c myc/p16/p21/p107/Rb cell cycle-related protein levels. A relevant increase in p27 protein levels was observed in anti-TGF-beta1-treated CLRPP, suggesting a role for p27 in the regulation of the hematopoietic potential. The present study on human progenitors and previously reported data on TGF-beta1 knockout mice suggest that, at the autocrine level, the cell cycle inhibitor TGF-beta1 plays an important role in regulating the survival and differentiation of primitive proliferating hematopoietic progenitors by cell cycle-independent mechanisms. PMID- 10807763 TI - Sustaining the graft-versus-tumor effect through posttransplant immunization with granulocyte-macrophage colony-stimulating factor (GM-CSF)-producing tumor vaccines. AB - For many cancers, autologous bone marrow transplantation (BMT) achieves a minimal residual disease state, yet relapse rates remain high. Using a syngeneic murine bone marrow transplant model, we demonstrate that vaccination with irradiated granulocyte-macrophage colony-stimulating factor (GM-CSF)-producing autologous tumor cells is effective in the post-BMT period and actually results in a greater tumor-free survival than vaccination in the nontransplant setting. Employing T cells specific for a model tumor-antigen, we find that transplantation of the tumor-bearing host results in a massive expansion and activation of tumor specific T cells in the early posttransplant period, but this response rapidly declines in association with tumor progression. Immunization with irradiated GM CSF tumor cells during the period of immune reconstitution results in the sustained amplification and activation of this response that closely correlates with freedom from relapse. These results demonstrate the feasibility of integrating GM-CSF vaccines in the postautologous BMT setting and suggest mechanisms that may contribute to the observed efficacy of immunization during the critical period of immune reconstitution. PMID- 10807764 TI - Detection of clonal T-cell receptor gamma-chain gene rearrangements in Reed Sternberg cells of classic Hodgkin disease. AB - Recent molecular single-cell studies have shown that in approximately 95% of cases, Reed-Sternberg cells of classic Hodgkin disease (HD) are derived from B cells of germinal center origin. Attempts to determine the cellular nature of the remaining cases have so far failed. To clarify whether they are derived from T cells, this study examined 791 single CD30(+) Hodgkin and Reed-Sternberg (HRS) cells from 13 T-cell marker-positive cases and from 6 cases with null-cell phenotype for rearranged T-cell receptor-gamma (TCR-gamma) genes by single copy polymerase chain reaction. Monoclonally rearranged TCR-gamma genes were detectable in 2 of the 13 classic HD cases with T-cell marker-positive HRS cells, with none detectable in the null-cell cases. Eight of the T-cell marker-positive cases and all 6 null-cell cases were also studied for rearrangements of immunoglobulin genes. Six of the 8 T-cell marker-positive cases harbored clonal immunoglobulin gene rearrangements. The 2 cases without rearranged immunoglobulin genes were those that contained clonal TCR-gamma rearrangements and lacked expression of the B-cell-specific activator protein. From these findings we conclude that cases of classic HD with T-cell-derived HRS cells definitely exist, although their overall incidence at 1% to 2% is very low. Even within the T-cell marker-positive cases only a minority (15%) were derived from T cells. The majority (85%) originated from B cells, indicating that the T-cell antigens expressed by HRS cells are, in contrast to those expressed in non-Hodgkin lymphoma, not lineage specific. PMID- 10807765 TI - Expression of the G-CSF receptor on hematopoietic progenitor cells is not required for their mobilization by G-CSF. AB - The mechanisms that regulate hematopoietic progenitor cell (HPC) mobilization from the bone marrow to blood have not yet been defined. HPC mobilization by granulocyte colony-stimulating factor (G-CSF), cyclophosphamide (CY), or interleukin-8 but not flt-3 ligand is markedly impaired in G-CSF receptor deficient (G-CSFR-deficient) mice. G-CSFR is expressed on mature hematopoietic cells, HPCs, and stromal cells, which suggests that G-CSFR signals in one or more of these cell types was required for mobilization by these agents. To define the cell type(s) responsible for G-CSF-dependent mobilization, a series of chimeric mice were generated using bone marrow transplantation. Mobilization studies in these chimeras demonstrated that expression of the G-CSFR on transplantable hematopoietic cells but not stromal cells is required for CY- or G-CSF-induced mobilization. Moreover, in irradiated mice reconstituted with both wild type and G-CSFR-deficient bone marrow cells, treatment with CY or G-CSF resulted in the equal mobilization of both types of HPCs. This result held true for a broad spectrum of HPCs including colony-forming cells, CD34(+) lineage(-) and Sca(+) lineage(-) cells, and long-term culture initiating cells. Collectively, these data provide the first definitive evidence that expression of the G-CSFR on HPCs is not required for their mobilization by G-CSF and suggest a model in which G CSFR-dependent signals act in trans to mobilize HPCs from the bone marrow. PMID- 10807766 TI - Chemokine receptors and their role in inflammation and infectious diseases. AB - Chemokines are small peptides that are potent activators and chemoattractants for leukocyte subpopulations and some nonhemopoietic cells. Their actions are mediated by a family of 7-transmembrane G-protein-coupled receptors, the size of which has grown considerably in recent years and now includes 18 members. Chemokine receptor expression on different cell types and their binding and response to specific chemokines are highly variable. Significant advances have been made in understanding the regulation of chemokine receptor expression and the intracellular signaling mechanisms used in bringing about cell activation. Chemokine receptors have also recently been implicated in several disease states including allergy, psoriasis, atherosclerosis, and malaria. However, most fascinating has been the observation that some of these receptors are used by human immunodeficiency virus type 1 in gaining entry into permissive cells. This review will discuss structural and functional aspects of chemokine receptor biology and will consider the roles these receptors play in inflammation and in infectious diseases. PMID- 10807767 TI - Rho proteins and the p38-MAPK pathway are important mediators for LPS-induced interleukin-8 expression in human endothelial cells. AB - Bacterial endotoxin (lipopolysaccharide, or LPS) has potent proinflammatory properties by acting on many cell types, including endothelial cells. Secretion of the CXC-chemokine interleukin-8 (IL-8) by LPS-activated endothelial cells contributes substantially to the inflammatory response. Using human umbilical vein endothelial cells (HUVECs), we analyzed the role of small GTP-binding Rho proteins and p38 mitogen-activated protein kinase (MAPK) for LPS-dependent IL-8 expression in endothelial cells. Specific inactivation of RhoA/Cdc42/Rac1 by Clostridium difficile toxin B-10463 (TcdB-10463) reduced LPS-induced tyrosine phosphorylation, nuclear factor (NF)-kappaB-dependent gene expression, IL-8 messenger RNA, and IL-8 protein accumulation but showed no effect on LPS dependent p38 MAPK activation. Inhibition of p38 MAPK by SB 202190 also blocked LPS-induced NF-kappaB activation and IL-8 synthesis. Furthermore, selective activation of the p38 MAPK pathway by transient expression of a constitutively active form of MAPK kinase (MKK)6, the upstream activator of p38, was as effective as LPS with respect to IL-8 expression in HUVECs. In summary, our data suggest that LPS-induced NF-kappaB activation and IL-8 synthesis in HUVECs are regulated by both a Rho-dependent signaling pathway and the MKK6/p38 kinase cascade. PMID- 10807768 TI - Rituximab monoclonal antibody as initial systemic therapy for patients with low grade non-Hodgkin lymphoma. AB - Rituximab, a chimeric antibody that targets CD20(+) B cells, produces a 48% response rate in patients with refractory low-grade non-Hodgkin lymphoma. In this phase II trial, patients with low-grade non-Hodgkin lymphoma who had previously received no systemic therapy were treated with rituximab, 375 mg/m(2), administered by IV infusion for 4 consecutive weeks. Patients with objective response or stable disease received repeat 4-week courses of rituximab at 6-month intervals. At the time of initial reevaluation at 6 weeks, 21 of 39 patients (54%) had objective response to treatment, and an additional 14 patients (36%) had stable disease or minor response. Response rates were similar in patients with follicular and small lymphocytic (CLL-type) lymphoma (52% versus 57%, respectively). At present, follow-up is short and only 13 patients have undergone a second course of rituximab treatment. However, 4 additional responses were documented either prior to the second course of rituximab (2 patients) or following the second course (2 patients) and 4 patients improved from partial to complete response. The current response rate is 64%, with 6 complete responses (15%). Treatment with rituximab was well tolerated, with only 1 patient experiencing grade 3/4 infusion-related toxicity. Rituximab is well tolerated and highly active in patients with low-grade non-Hodgkin lymphoma previously untreated with systemic therapy. Although further follow-up is required, the demonstration of minimal toxicity and considerable activity of this new biologic agent represents an important beginning of more specific, less toxic treatment for this important group of cancer patients. PMID- 10807769 TI - Bone marrow transplantation versus periodic prophylactic blood transfusion in sickle cell patients at high risk of ischemic stroke: a decision analysis. AB - Measurement of cerebral blood velocity (CBV) by transcranial Doppler has been used to identify patients with sickle cell disease (SCD) who are at high risk of ischemic stroke. This study examines outcomes of bone marrow transplantation (BMT) and periodic blood transfusion (PBT) as a basis for making treatment recommendations for patients who have elevated CBV and no other indications for BMT. Decision analysis was used to compare the number of quality-adjusted life years (QALYs) experienced by a population of patients with SCD at high risk for stroke who were treated with PBT or BMT. Markov models were constructed to represent the clinical course of patients with SCD who were treated with PBT or BMT. Medical literature and expert opinion provided risks of stroke and death for different disease states, estimates of transition probabilities from one clinical state to another, and quality of life. An intention-to-treat analysis and an analysis of treatment received were both performed on hypothetical cohorts of 100 000 patients. Patients with SCD who were managed with a strategy of intending to provide BMT could expect 16.0 QALYs, compared with 15.7 QALYs for a strategy of intending to provide PBT; however, the variation around these estimates was large. In the treatment received analysis, patients compliant with PBT therapy and iron chelation could expect the best outcomes (19.2 QALYs). From a policy perspective, neither BMT nor PBT can be considered the "best" treatment for children with SCD who have abnormal CBV. Abnormal CBV should not be the only criterion for selecting patients with sickle cell for BMT. PMID- 10807771 TI - Stable and functional lymphoid reconstitution of common cytokine receptor gamma chain deficient mice by retroviral-mediated gene transfer. AB - Mutations in the gene encoding the common cytokine receptor gamma chain (gamma(c)) are responsible for human X-linked severe combined immunodeficiency disease (SCIDX1). We have used a gamma(c)-deficient mouse model to test the feasibility and potential toxicity of gamma(c) gene transfer as a therapy for SCIDX1. A retrovirus harboring the murine gamma(c) chain was introduced into gamma(c)-deficient bone marrow cells, which were then transplanted into alymphoid RAG2/gamma(c) double-deficient recipient mice. Circulating lymphocytes appeared 4 weeks postgraft and achieved steady-state levels by 8 weeks. The mature lymphocytes present in the grafted mice had integrated the gamma(c) transgene, expressed gamma(c) transcripts, and were able to proliferate in response to gamma(c)-dependent cytokines. The gamma(c)-transduced animals demonstrated (1) normal levels of immunoglobulin subclasses, including immunoglobulin G1 (IgG1) and IgG2a (which are severely decreased in gamma(c)(-) mice); (2) the ability to mount an antigen-specific, T-dependent antibody response showing effective in vivo T-B cell cooperation, and (3) the presence of gut-associated cryptopatches and intraepithelial lymphocytes. Importantly, peripheral B and T cells were still present 47 weeks after a primary graft, and animals receiving a secondary graft of gamma(c)-transduced bone marrow cells demonstrated peripheral lymphoid reconstitution. That gamma(c) gene transfer to hematopoietic precursor cells can correct the immune system abnormalities in gamma(c)(-) mice supports the feasibility of in vivo retroviral gene transfer as a treatment for human SCIDX1. PMID- 10807770 TI - Hepatitis C infection among survivors of childhood cancer. AB - Preliminary reports have suggested that survivors of childhood cancer and aplastic anemia who are infected with the hepatitis C virus (HCV) have a low risk for progression to significant liver disease. Among our surviving patients who were transfused between 1961 and March 1992, 77 (6.6% of surviving patients tested thus far) have evidence of HCV infection, whereas 4 surviving patients who were transfused after March 1992 are HCV-infected. One patient chronically infected with HCV died of liver failure, and 2 patients died of hepatocellular carcinoma. To characterize the risk for these and other complications, 65 patients are enrolled in a longitudinal study of HCV infection, of whom 58 (89.2%) had circulating HCV RNA at the time of protocol enrollment, with genotypes 1A and 1B most commonly isolated. Most enrolled patients have few or no symptoms, carry out normal activities, and have normal liver function. To date, 35 patients have undergone liver biopsy for abnormal liver function since the diagnosis of primary malignancy; central pathology review shows 28 (80%) have chronic active hepatitis, 25 (71%) have fibrosis, and 3 (9%) have cirrhosis. These preliminary data suggest that though most survivors of childhood cancer who are infected with HCV are clinically well, some are at risk for clinically significant liver disease. Identification of other HCV-infected patients and prospective monitoring of this cohort are ongoing to determine the risk for, and to identify factors associated with the progression of, liver disease. PMID- 10807772 TI - Limiting numbers of G156A O(6)-methylguanine-DNA methyltransferase-transduced marrow progenitors repopulate nonmyeloablated mice after drug selection. AB - The limited efficacy of hematopoietic gene therapy can be improved by in vivo selection for transduced long-term repopulating cells (LTRC). We selected for G156A MGMT (triangle upMGMT) transduced LTRC present in 5 x 10(4) to 100 x 10(4) marrow cells infused into nonmyeloablated mice by the administration of O(6) benzylguanine (BG) and BCNU every 3 to 4 weeks. To facilitate engraftment, mice were given a nonablative dose of BG and BCNU before infusion. Without selection, triangle upMGMT was not detected in any hematopoietic colony-forming units (CFU) 24 to 30 weeks after infusion. After BG and BCNU, triangle upMGMT(+) CFU were frequently detected, and their proportions increased with each treatment cycle. After 2 to 3 cycles of BG and BCNU, many mice were stably reconstituted with 75% to 100% triangle upMGMT(+) CFU for at least 6 months, representing up to 940-fold enrichment. Thus, BG and BCNU stem cell toxicity allows triangle upMGMT transduced LTRC to repopulate the bone marrow. This degree of selection pressure in nonmyeloablated mice is far greater than that observed in previous drug resistance gene transfer studies. These data support our approved clinical trial to select for drug-resistant, transduced hematopoietic cells, potentially decreasing cumulative drug-induced myelosuppression in patients with cancer. These data also suggest that triangle upMGMT may be a potent, dominant, selectable marker for use in dual gene therapy. PMID- 10807773 TI - Efficient transduction of human hematopoietic repopulating cells generating stable engraftment of transgene-expressing cells in NOD/SCID mice. AB - In an attempt to develop efficient procedures of human hematopoietic gene therapy, retrovirally transduced CD34(+) cord blood cells were transplanted into NOD/SCID mice to evaluate the repopulating potential of transduced grafts. Samples were prestimulated on Retronectin-coated dishes and infected with gibbon ape leukemia virus (GALV)-pseudotyped FMEV vectors encoding the enhanced green fluorescent protein (EGFP). Periodic analyses of bone marrow (BM) from transplanted recipients revealed a sustained engraftment of human hematopoietic cells expressing the EGFP transgene. On average, 33.6% of human CD45(+) cells expressed the transgene 90 to 120 days after transplantation. Moreover, 11.9% of total NOD/SCID BM consisted of human CD45(+) cells expressing the EGFP transgene at this time. The transplantation of purified EGFP(+) cells increased the proportion of CD45(+) cells positive for EGFP expression to 57. 7% at 90 to 120 days after transplantation. At this time, 18.9% and 4.3% of NOD/SCID BM consisted of CD45(+)/EGFP(+) and CD34(+)/EGFP(+) cells, respectively. Interestingly, the transplantation of EGFP(-) cells purified at 24 hours after infection also generated a significant engraftment of CD45(+)/EGFP(+) and CD34(+)/EGFP(+) cells, suggesting that a number of transduced repopulating cells did not express the transgene at that time. Molecular analysis of NOD/SCID BM confirmed the high levels of engraftment of human transduced cells deduced from FACS analysis. Finally, the analysis of the provirus insertion sites by conventional Southern blotting indicated that the human hematopoiesis in the NOD/SCID BM was predominantly oligoclonal. PMID- 10807774 TI - The role of platelet alpha-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells. AB - Thrombopoietin (TPO), the specific cytokine that regulates platelet production, is expressed in human bone marrow (BM), kidney, and liver. There appears to be no regulation of TPO in the kidney and liver, but TPO messenger RNA (mRNA) expression can be modulated in the stromal cells of the BM. In this study, we used primary human BM stromal cells as a model to study the regulation of TPO mRNA expression in response to various platelet alpha-granular proteins. We showed that platelet-derived growth factor (PDGF) BB and fibroblast growth factor (FGF) 2 stimulated TPO mRNA expression in both a dose-dependent and time dependent manner. The addition of 50 ng/mL of PDGF and 20 ng/mL of FGF resulted in maximal induction of TPO mRNA expression in 4 hours. We also found that platelet factor 4 (PF4), thrombospondin (TSP), and transforming growth factor beta (TGF-beta) are negative modulators of megakaryocytopoiesis. We observed suppression in TPO mRNA expression with 1 microg/mL of both PF4 and TSP and 50 ng/mL of TGF-beta, with maximal suppression occurring 4 hours after the addition of these proteins. Finally, the addition of whole-platelet lysate produced a dose dependent inhibition of TPO expression. On the basis of these findings, we propose that the platelet alpha-granular proteins studied may regulate TPO gene expression in BM stromal cells by means of a feedback mechanism. PMID- 10807775 TI - beta2 microglobulin-deficient (B2m(null)) NOD/SCID mice are excellent recipients for studying human stem cell function. AB - Human SCID repopulating cells (SRC) are defined based on their functional ability to repopulate the bone marrow of NOD/SCID mice with both myeloid and lymphoid cell populations. The frequency of SRC in umbilical cord blood cells is 1 in 9.3 x 10(5) mononuclear cells. We report that as few as 8 x 10(4) human cord blood mononuclear cells transplanted into NOD/SCID/B2m(null )mice resulted in multilineage differentiation in the murine bone marrow, revealing a more than 11 fold higher SRC frequency than in NOD/SCID mice. Moreover, as few as 2 to 5 x 10(3) CD34(+) cells recovered from the bone marrow of primary transplanted NOD/SCID mice were sufficient for engrafting secondary NOD/SCID/B2m(null )mice with SRC, suggesting SRC self-renewal. Thus, by using NOD/SCID/B2m(null )mice as recipients, we established a functional assay for human stem cells capable of engrafting the bone marrow of primary and secondary transplanted immune-deficient mice with SRC, providing a model that better resembles autologous stem cell transplantation. PMID- 10807776 TI - In vitro differentiation of endothelial cells from AC133-positive progenitor cells. AB - Recent findings support the hypothesis that the CD34(+)-cell population in bone marrow and peripheral blood contains hematopoietic and endothelial progenitor and stem cells. In this study, we report that human AC133(+) cells from granulocyte colony-stimulating factor-mobilized peripheral blood have the capacity to differentiate into endothelial cells (ECs). When cultured in the presence of vascular endothelial growth factor (VEGF) and the novel cytokine stem cell growth factor (SCGF), AC133(+) progenitors generate both adherent and proliferating nonadherent cells. Phenotypic analysis of the cells within the adherent population reveals that the majority display endothelial features, including the expression of KDR, Tie-2, Ulex europaeus agglutinin-1, and von Willebrand factor. Electron microscopic studies of these cells show structures compatible with Weibel-Palade bodies that are found exclusively in vascular endothelium. AC133 derived nonadherent cells give rise to both hematopoietic and endothelial colonies in semisolid medium. On transfer to fresh liquid culture with VEGF and SCGF, nonadherent cells again produce an adherent and a nonadherent population. In mice with severe combined immunodeficiency, AC133-derived cells form new blood vessels in vivo when injected subcutaneously together with A549 lung cancer cells. These data indicate that the AC133(+)-cell population consists of progenitor and stem cells not only with hematopoietic potential but also with the capacity to differentiate into ECs. Whether these hematopoietic and endothelial progenitors develop from a common precursor, the hemangioblast will be studied at the single-cell level. PMID- 10807777 TI - Functionally defined CD164 epitopes are expressed on CD34(+) cells throughout ontogeny but display distinct distribution patterns in adult hematopoietic and nonhematopoietic tissues. AB - Three distinct classes of epitopes on human CD164 have been identified. Two of these, recognized by the monoclonal antibodies 105A5 and 103B2/9E10, are the CD164 class I and class II functionally defined epitopes, which cooperate to regulate adhesion and proliferation of CD34(+) cell subsets. In this article, we demonstrate that these 2 CD164 epitopes are expressed on CD34(+) cells throughout ontogeny, in particular on CD34(+ )cell clusters associated with the ventral floor of the dorsal aorta in the developing embryo and on CD34(+) hematopoietic precursor cells in fetal liver, cord blood, and adult bone marrow. While higher levels of expression of these CD164 epitopes occur on the more primitive AC133(hi)CD34(hi)CD38(lo/-) cell population, they also occur on most cord blood Lin(-)CD34(lo/-)CD38(lo/- )cells, which are potential precursors for the AC133(hi)CD34(hi)CD38(lo/-) subset. In direct contrast to these common patterns of expression on hematopoietic precursor cells, notable differences in expression of the CD164 epitopes were observed in postnatal lymphoid and nonhematopoietic tissues, with the class I and class II CD164 epitopes generally exhibiting differential and often reciprocal cellular distribution patterns. This is particularly striking in the colon, where infiltrating lymphoid cells are CD164 class I-positive but class II-negative, while epithelia are weakly CD164 class II positive. Similarly, in certain lymphoid tissues, high endothelial venules and basal and subcapsular epithelia are CD164 class II-positive, while lymphoid cells are CD164 class I-positive. It therefore seems highly likely that these CD164 class I and II epitopes will mediate reciprocal homing functions in these tissue types. PMID- 10807778 TI - Characterization of Ly-6M, a novel member of the Ly-6 family of hematopoietic proteins. AB - The Ly-6 family includes a number of highly homologous, low molecular weight glycophosphatidylinositol-linked proteins expressed on hematopoietic and lymphoid cells. The best characterized family member is Sca-1 (Ly-6A/E), an antigen commonly used for purification of murine pluripotent hematopoietic cells. We sought to characterize the genomic locus surrounding the Sca-1 gene. We identified several overlapping P1 artificial chromosomes containing the Sca-1 gene and mapped one of these to mouse chromosome 15D3.1-3.3, the region previously shown to contain members of the murine Ly-6 gene family. We then mapped this clone and found that the Sca-2 gene lies 35.4 kilobase (kb) downstream of Sca-1 in the opposite transcriptional orientation. This is the first direct demonstration of physical linkage of Ly-6 genes. A novel gene, highly homologous to Sca-1 was identified and localized 13.4 kb downstream of Sca 1. This gene, which we designated Ly-6M, shares several structural features conserved among members of the Ly-6 family. Ly-6M messenger RNA (mRNA) is easily detectable in hematopoietic tissue (bone marrow, spleen, thymus, peritoneal macrophages) as well as kidney and lung. No mRNA expression was detected in heart, stomach, liver, small intestine, brain, or skin. Ly-6M protein is detectable on 10% to 15% of peripheral blood leukocytes, including monocytes and a subpopulation of B220(+) cells. Ly-6M is broadly distributed in the bone marrow, with prominent expression on monocytes and myeloid precursors. The identification and characterization of Ly-6M adds a new member to a complex family of homologous, tightly linked genes that have proven extremely useful reagents for defining populations within the hematopoietic system. PMID- 10807779 TI - Concentration-dependent dual effect of thrombin on impaired growth/apoptosis or mitogenesis in tumor cells. AB - Because thrombin-treated tumor cell-induced metastasis increases tumor nodule volume(12) greater than nodule number, we studied the effect of thrombin on tumor cell growth in vitro and in vivo (murine B16F10 melanoma, human HCT8 colon carcinoma, DU145 prostate carcinoma). Tumor cell growth was measured after 3 to 7 days in 1% fetal calf serum (FCS) + RPMI 1640. We found that, whereas relatively low concentrations of thrombin, 0.1 to 0.5 U/mL (1-5 nmol/L) enhance tumor cell growth in vitro approximately 2- to 3-fold, higher concentrations, 0.5 to 1 U/mL (5-10 nmol/L) impaired cell growth approximately 2- to 4-fold. Impaired cell growth was associated with cell cycle arrest at G(2)M and increased pre-G(o) DNA, as well as apoptosis, measured by tumor cell binding to Annexin V and propidium iodide. Apoptosis was reversed with the general caspase inhibitor, FK-011. The enhancing and inhibiting effects were specific for thrombin (reversed with inactive diisopropyl-fluorophosphate [DFP]-thrombin) and mediated via the protease-activated receptor 1 (PAR-1). PAR-1 activation was demonstrated by (1) use of a cell line, B16F10, devoid of the 3 other thrombin receptors, PAR-3, PAR 4, and GPIb; and (2) greater sensitivity of PAR-1 transfected B16F10 and HCT8 cells to impaired cell growth/apoptosis, 3- and 14-fold, respectively. Thus, thrombin has a bimodal effect on PAR-1 in tumor cells: enhanced growth at low concentration, impaired growth/apoptosis at higher concentration. PMID- 10807780 TI - Conformational changes in the D' domain of von Willebrand factor induced by CYS 25 and CYS 95 mutations lead to factor VIII binding defect and multimeric impairment. AB - We report 2 new mutations identified in 3 patients and characterized by the markedly decreased affinity of von Willebrand factor (vWF) for factor VIII (FVIII). Patients 2 and 3, who have a typical type 2N phenotype, were found to be compound heterozygous for Arg91Gln and Cys25Tyr or Cys95Phe, respectively. Patient 1, who is the first cousin of patient 2, had an FVIII binding defect of vWF, low levels of vWF, and multimeric impairment. She was found to be compound heterozygous for the mutations Cys25Tyr and a stop codon (D93ter) in exon 4. Transient expression of recombinant vWF (rvWF) containing either Cys25Tyr or Cys95Phe mutations resulted in mutated rvWF with markedly reduced FVIII binding ability, multimeric structure impairment, and a significant decrease in the vWF expression level. Moreover, the use of anti-vWF monoclonal antibodies that inhibit the FVIII binding showed that these 2 mutations likely induce a conformational change in the D' domain. These results show that the native conformation of the D' domain of vWF is not only required for FVIII binding but also for normal multimerization and optimal secretion. PMID- 10807781 TI - Interleukin-13 induces PSGL-1/P-selectin-dependent adhesion of eosinophils, but not neutrophils, to human umbilical vein endothelial cells under flow. AB - Selective eosinophil accumulation is a hallmark of diseases such as asthma. In a model of chronic eosinophilic inflammation, we have previously shown that the tethering step in eosinophil adhesion is mediated by PSGL-1 binding to P selectin. The Th2-associated cytokine IL-13 is of potential importance in allergic disease. We have therefore investigated whether IL-13 can mediate eosinophil binding to human umbilical vein endothelial cells (HUVEC) through P selectin. IL-13 caused dose- and time-dependent increases of P-selectin expression, as assessed by flow and laser scanning cytometry. A similar degree of expression was observed with IL-4. There was no effect on E-selectin or ICAM-1 expression. Tumor necrosis factor-alpha induced the expression of VCAM-1, E selectin, and ICAM-1 but had no effect on P-selectin expression. IL-13 increased the production of mRNA for surface and soluble variants of P-selectin. Under flow conditions, eosinophils, but not neutrophils, showed enhanced binding to IL-13 and to IL-4-stimulated HUVEC compared to medium-cultured cells. Eosinophil adhesion was completely inhibited by a blocking monoclonal antibody against PSGL 1 and P-selectin. Anti-VLA-4 and anti-VCAM-1 antibodies inhibited binding to a lesser extent. Thus, at physiologic levels of expression induced by Th2 cytokines, P-selectin/PSGL-1 supported eosinophil but not neutrophil adhesion. This mechanism is likely to be a key event leading to the selective accumulation of eosinophils in allergic inflammation. PMID- 10807782 TI - Blockade of CD86 and CD40 induces alloantigen-specific immunoregulatory T cells that remain anergic even after reversal of hyporesponsiveness. AB - The generation of immunoregulatory T cells that block the B7(CD86/CD80)-CD28 and/or CD40-CD154 costimulatory pathways has great potential for the induction of long-term transplantation tolerance. In a human polyclonal in vitro model, combined monoclonal antibody (mAb) blocking of the costimulatory ligands CD40 and CD86 lead to allospecific T-cell anergy that cannot be reversed by antigenic rechallenge in the presence of IL-2. Although antigenic restimulation with IL-2 restored the proliferative response, subsequent antigenic restimulation of the restored anergic cells in a tertiary mixed lymphocyte culture still resulted in nonresponsiveness. Importantly, these anergic T cells suppress the response of naive alloreactive T cells in an antigen-specific way via linked recognition. Suppression may partially depend on local IL-10 production, while transforming growth factor-beta (TGF-beta) did not play a role. Irrespective of the monoclonal antibody combination used, blast formation occurred in a subset of CD4(+) cells. These cells were characterized by a sustained CD45RA expression, an increased T cell receptor density, and a lower level of CD4 expression. A reduced number of CD45RO(+)/CD8(+) T cells was observed whenever anti-CD86 was combined with anti CD40, which was reflected by an even more attenuated cytotoxic T-cell function. This indicates the importance of CD40-CD154 in the generation of cytotoxic T cells in this transplantation model. We hypothesize that in our model, anergy is induced in the CD4(+) T-cell subset, whereby CD8(+) cytotoxic effector function is impaired by the lack of both CD40-CD154 signaling and cytokine-mediated help. This costimulatory ligand-directed mAb approach might well be used for the ex vivo generation of antigen-specific immunoregulatory T cells applicable in adoptive immunotherapy. PMID- 10807783 TI - Glucocorticoids transform CD40-triggering of dendritic cells into an alternative activation pathway resulting in antigen-presenting cells that secrete IL-10. AB - Dendritic cell (DC) activation through CD40-CD40 ligand interactions is a key regulatory step for the development of protective T-cell immunity and also plays an important role in the initiation of T-cell responses involved in autoimmune diseases and allograft rejection. In contrast to previous reports, we show that the immunosuppressive drug dexamethasone (DEX) redirects rather than simply blocks this DC activation process. We found that DCs triggered through CD40 in the presence of DEX were unable to acquire high levels of costimulatory, adhesion, and major histocompatibility complex class I and II molecules and failed to express the maturation marker CD83, whereas antigen uptake was not affected. Moreover, DEX strikingly modified the CD40-activated DC cytokine secretion profile by suppressing the production of the proinflammatory cytokine interleukin (IL)-12 and potentiating the secretion of the anti-inflammatory cytokine IL-10. Accordingly, DEX-exposed CD40-triggered DCs displayed a decreased T-cell allostimulatory potential and a dramatically impaired ability to activate cloned CD4(+) T helper 1 (Th1) cells. Moreover, interaction between Th1 cells and these DCs rendered the T cells hyporesponsive to further antigen-specific restimulation. Collectively, our results demonstrate that DEX profoundly modulates CD40-dependent DC activation and suggest that the resulting alternatively activated DCs can be exploited for suppression of unwanted T-cell responses in vivo. PMID- 10807784 TI - Thrombocyte HLA molecules retain nonrenewable endogenous peptides of megakaryocyte lineage and do not stimulate direct allocytotoxicity in vitro. AB - The origin and the function of HLA class I molecules present on the surface of human platelets are still unclear. In particular, it is controversial which fraction of these class I molecules represents integral membrane components derived from the megakaryocyte-platelet lineage versus soluble plasma HLA molecules acquired by adsorption. Results of the present study show that HLA-A2 ligands isolated from platelets possess the same peptide motif as described for HLA-A2-associated peptides obtained from nucleated cells. Sequencing of these platelet-derived peptides reveals that they originate mainly from ubiquitously expressed proteins also present in the megakaryocyte-platelet lineage. Moreover, one of these peptides derives from the GPIX protein, which is specifically expressed by platelets and their precursors. Platelet HLA molecules are unstable in vitro at 37 degrees C, but can be partially stabilized by addition of exogenous beta(2)-microglobulin and HLA class I binding peptide, suggesting that platelets cannot load HLA molecules with endogenous peptides. In in vitro experiments platelets were used to stimulate peripheral blood mononuclear cells. No allospecific cytotoxicity was observed after primary stimulation, or secondary restimulation, with allogenic resting or activated platelets, even in the presence of additional third-party helper activity. These data indicate that HLA class I molecules from platelets cannot directly induce allogenic CD8(+) cytotoxic T-cell response in vitro. PMID- 10807785 TI - Deficiency of the Fas apoptosis pathway without Fas gene mutations is a familial trait predisposing to development of autoimmune diseases and cancer. AB - Fas/Apo-1 (CD95) triggers programmed cell death (PCD) and is involved in immune response control and cell-mediated cytotoxicity. In the autoimmune/lymphoproliferative syndrome (ALPS), inherited loss-of-function mutations of the Fas gene cause nonmalignant lymphoproliferation and autoimmunity. We have recently identified an ALPS-like clinical pattern (named autoimmune lymphoproliferative disease [ALD]) in patients with decreased Fas function, but no Fas gene mutation. They also displayed decreased PCD response to ceramide, triggering a death pathway partially overlapping that used by Fas, which suggests that ALD is caused by downstream alterations of the Fas signaling pathway. Decreased Fas function is also involved in tumor development, because somatic mutations hitting the Fas system may protect neoplastic cells from immune surveillance. This work assessed the inherited component of the ALD defect by evaluating Fas- and ceramide-induced T-cell death in both parents and 4 close relatives of 10 unrelated patients with ALD. Most of them (22 of 24) displayed defective Fas- or ceramide-induced (or both) cell death. Moreover, analysis of the family histories showed that frequencies of autoimmunity and cancer were significantly increased in the paternal and maternal line, respectively. Defective Fas- or ceramide-induced T-cell death was also detected in 9 of 17 autoimmune patients from 7 families displaying more than a single case of autoimmunity within first- or second-degree relatives (multiple autoimmune syndrome [MAS] patients). Autoimmune diseases displayed by ALD and MAS families included several organ-specific and systemic forms. These data suggest that ALD is due to accumulation of several defects in the same subject and that these defects predispose to development of cancer or autoimmune diseases other than ALPS/ALD. PMID- 10807786 TI - Interleukin-2 enhances the response of natural killer cells to interleukin-12 through up-regulation of the interleukin-12 receptor and STAT4. AB - Interleukin (IL)-12 plays a critical role in modulating the activities of natural killer (NK) cells and T lymphocytes. In animal models, IL-12 has potent antitumor effects that are likely mediated by its ability to enhance the cytotoxic activity of NK cells and cytotoxic T lymphocytes, and to induce the production of interferon (IFN)-gamma by NK and T cells. In addition to IL-12, NK cells are responsive to IL-2, and may mediate some of the antitumor effects of IL-2. In this study, we examine the interaction between IL-2 and the signaling events induced by IL-12 in NK cells. We find that IL-2 not only up-regulates the expression of IL-12Rbeta1 and IL-12Rbeta2, it also plays an important role in up regulating and maintaining the expression of STAT4, a critical STAT protein involved in IL-12 signaling in NK cells. In contrast to the effects of IL-2 alone, expression of IL-12 receptors and STAT4 are unaffected or decreased by IL 12 or the combination of IL-2 and IL-12. Through expression of high levels of IL 12 receptors and STAT4, IL-2-primed NK cells show enhanced functional responses to IL-12 as measured by IFN-gamma production and the killing of target cells. NK cells from cancer patients who received low-dose IL-2 treatment also exhibited increased expression of IL-12 receptor chains, suggesting that IL-2 may enhance the response to IL-12 in vivo. These findings provide a molecular framework to understand the interaction between IL-2 and IL-12 in NK cells, and suggest strategies for improving the effectiveness of these cytokines in the immunotherapy of cancer. PMID- 10807787 TI - Alteration of tumor necrosis factor-alpha T-cell homeostasis following potent antiretroviral therapy: contribution to the development of human immunodeficiency virus-associated lipodystrophy syndrome. AB - Highly-active antiretroviral therapy (HAART) has lead to a dramatic decrease in the morbidity of patients infected with the human immunodeficiency virus (HIV). However, metabolic side effects, including lipodystrophy-associated (LD associated) dyslipidemia, have been reported in patients treated with antiretroviral therapy. This study was designed to determine whether successful HAART was responsible for a dysregulation in the homeostasis of tumor necrosis factor-alpha (TNF-alpha), a cytokine involved in lipid metabolism. Cytokine production was assessed at the single cell level by flow cytometry after a short term stimulation of peripheral blood T cells from HIV-infected (HIV(+)) patients who were followed during 18 months of HAART. A dramatic polarization to TNF-alpha synthesis of both CD4 and CD8 T cells was observed in all patients. Because it was previously shown that TNF-alpha synthesis by T cells was highly controlled by apoptosis, concomitant synthesis of TNF-alpha and priming for apoptosis were also analyzed. The accumulation of T cells primed for TNF-alpha synthesis is related to their escape from activation-induced apoptosis, partly due to the cosynthesis of interleukin-2 (IL-2) and TNF-alpha. Interestingly, we observed that LD is associated with a more dramatic TNF-alpha dysregulation, and positive correlations were found between the absolute number of TNF-alpha CD8 T-cell precursors and lipid parameters usually altered in LD including cholesterol, triglycerides, and the atherogenic ratio apolipoprotein B (apoB)/apoA1. Observations from the study indicate that HAART dysregulates homeostasis of TNF alpha synthesis and suggest that this proinflammatory response induced by efficient antiretroviral therapy is a risk factor of LD development in HIV(+) patients. PMID- 10807788 TI - RasGRP links T-cell receptor signaling to Ras. AB - Stimulation of the T-cell receptor (TCR) alters a number of intracellular signaling pathways including one that involves protein tyrosine kinases, phospholipase C-gamma1 (PLC-gamma1), diacylglycerol (DAG), and calcium messengers. By a divergent pathway, TCR-stimulated protein tyrosine kinase activity is thought to result independently in recruitment of the Ras activator Sos to the plasma membrane, leading to Ras activation. Here we show that RasGRP, a Ras activator that contains calcium-binding EF hands and a DAG-binding domain, is expressed in T cells. A PLC-gamma1 inhibitor diminished activation of Ras following TCR stimulation. Membranes from TCR-stimulated Jurkat T cells exhibited increased RasGRP and increased Ras-guanyl nucleotide association activity that was inhibited by antibodies directed against RasGRP. Overexpression of RasGRP in T cells enhanced TCR-Ras-Erk signaling and augmented interleukin-2 secretion in response to calcium ionophore plus DAG analogues phorbol ester myristate or bryostatin-1. Thus, RasGRP links TCR and PLC-gamma1 to Ras-Erk signaling, a pathway amenable to pharmacologic manipulation. PMID- 10807789 TI - Further demonstration of the diversity of chromosomal changes involving 2p23 in ALK-positive lymphoma: 2 cases expressing ALK kinase fused to CLTCL (clathrin chain polypeptide-like). AB - Anaplastic lymphoma kinase (ALK)-positive lymphomas are characterized by expression of a hybrid protein, comprising the cytoplasmic portion of the ALK tyrosine kinase fused to a partner protein. This hybrid kinase is often encoded by the nucleophosmin (NPM) NPM-ALK fusion gene resulting from the (2;5)(p23;q35) chromosomal translocation. However, the ALK gene at 2p23 may also be involved in 2 variant translocations, namely t(1;2)(q25;p23) and t(2;3)(p23;q21), which create the TPM3-ALK and TFG-ALK fusion genes, respectively. We report here 2 lymphomas with an unusual finely granular cytoplasmic ALK staining pattern, clearly different from the pattern observed in ALK-positive lymphomas carrying NPM-ALK or its variants. A cloned complementary DNA sequence from 1 of these 2 lymphomas contained the ALK gene fused to the second clathrin heavy chain gene (also referred to as clathrin heavy polypeptide-like gene) (CLTCL). The distinctive granular cytoplasmic staining pattern for ALK was likely to be due to binding of the fusion protein to clathrin-coated vesicles. The CLTCL gene is constitutively expressed in lymphoid cells and therefore presumably contributes an active promoter for the CLTCL-ALK gene. The fusion protein had a molecular weight (250 kd) that differs from all known ALK products, and it was autophosphorylated in an in vitro kinase assay, confirming that it is constitutively active and hence capable of contributing to malignant transformation. These 2 cases, therefore, represent a hitherto undescribed mechanism of ALK activation in lymphoma and further illustrate the diversity of fusion partners for the ALK gene. PMID- 10807790 TI - Hypermethylation of E-cadherin in leukemia. AB - E-cadherin gene is often termed a "metastasis suppressor" gene because the E cadherin protein can suppress tumor cell invasion and metastasis. Inactivation of the E-cadherin gene occurs in undifferentiated solid tumors by both genetic and epigenetic mechanisms; however, the role of E-cadherin in hematologic malignancies is only now being recognized. E-cadherin expression is essential for erythroblast and normoblast maturation, yet expression is reduced or absent in leukemic blast cells. This study examined the messenger RNA (mRNA) and protein expression of the E-cadherin gene in bone marrow and blood samples from normal donors and patients with leukemia. We found that all normal donor samples expressed E-cadherin mRNA, whereas both samples of acute myelogenous leukemia and chronic lymphocytic leukemia had a significant reduction or absence of expression. However, normal blast counterparts expressed only a low level of E cadherin surface protein. Sodium bisulphite genomic sequencing was used to fully characterize the methylation patterns of the CpG island associated with the E cadherin gene promoter in those samples with matched DNA. All of the normal control samples were essentially unmethylated; however, 14 of 18 (78%) of the leukemia samples had abnormal hypermethylation of the E-cadherin CpG island. In fact both alleles of the E-cadherin gene were often hypermethylated. We conclude the E-cadherin gene is a common target for hypermethylation in hematologic malignancies. PMID- 10807791 TI - Glyoxalase I is involved in resistance of human leukemia cells to antitumor agent induced apoptosis. AB - Abnormality in the machinery of apoptosis is associated with a resistant phenotype of the tumor cell to chemotherapy. To determine the molecular basis of resistance to antitumor agent-induced apoptosis, we performed a complementary DNA (cDNA) subtractive hybridization with messenger RNA (mRNA) from human monocytic leukemia U937 and its variant UK711, which is resistant to apoptosis induced by antitumor agents. We found that glyoxalase I (GLO1), an enzyme that detoxifies methylglyoxal, is selectively overexpressed in the apoptosis-resistant UK711 cells. The GLO1 enzyme activity was significantly elevated in UK711 and UK110 cells, another drug-resistant mutant, as well as in K562/ADM, adriamycin resistant leukemia cells, compared with their parental cells. When overexpressed in human Jurkat cells, GLO1 inhibited etoposide- and adriamycin-induced caspase activation and apoptosis, indicating the involvement of GLO1 in apoptosis suppression caused by these drugs. Moreover, cotreatment with S-p bromobenzylglutathione cyclopentyl diester (BBGC), a cell-permeable inhibitor of GLO1, enhanced etoposide-induced apoptosis in resistant UK711 cells but not in parental U937 cells. Taken together, these results indicate that GLO1 is a resistant factor to antitumor agent-induced apoptosis in human leukemia cells and that the GLO1 inhibitor could be a drug resistance-reversing agent. PMID- 10807792 TI - Chronic neutropenia mediated by fas ligand. AB - Chronic neutropenia, often associated with rheumatoid arthritis, is a characteristic finding in large granular lymphocyte (LGL) leukemia. The mechanism of neutropenia is not known. Normal neutrophil survival is regulated by the Fas Fas ligand apoptotic system. We hypothesized that neutropenia in LGL leukemia is mediated by dysregulated expression of Fas ligand. Levels of Fas ligand in serum samples from patients with LGL leukemia were measured with a Fas ligand enzyme linked immunosorbent assay. The effects of serum from patients with LGL leukemia on apoptosis of normal neutrophils were determined by flow cytometry and morphologic assessment. High levels of circulating Fas ligand were detected in 39 of 44 serum samples from patients with LGL leukemia. In contrast, Fas ligand was undetectable in 10 samples from healthy donors. Serum from the patients triggered apoptosis of normal neutrophils that depended partly on the Fas pathway. Resolution of neutropenia was associated with disappearance or marked reduction in Fas ligand levels in 10 of 11 treated patients. These data suggest that high levels of Fas ligand are a pathogenetic mechanism in human disease. (Blood. 2000;95:3219-3222) PMID- 10807793 TI - The gene for familial Mediterranean fever, MEFV, is expressed in early leukocyte development and is regulated in response to inflammatory mediators. AB - Familial Mediterranean fever (FMF) is a recessive disorder characterized by episodes of fever and neutrophil-mediated serosal inflammation. We recently identified the gene causing FMF, designated MEFV, and found it to be expressed in mature neutrophils, suggesting that it functions as an inflammatory regulator. To facilitate our understanding of the normal function of MEFV, we extended our previous studies. MEFV messenger RNA was detected by reverse transcriptase polymerase chain reaction in bone marrow leukocytes, with differential expression observed among cells by in situ hybridization. CD34 hematopoietic stem-cell cultures induced toward the granulocytic lineage expressed MEFV at the myelocyte stage, concurrently with lineage commitment. The prepromyelocytic cell line HL60 expressed MEFV only at granulocytic and monocytic differentiation. MEFV was also expressed in the monocytic cell lines U937 and THP-1. Among peripheral blood leukocytes, MEFV expression was detected in neutrophils, eosinophils, and to varying degrees, monocytes. Consistent with the tissue specificity of expression, complete sequencing and analysis of upstream regulatory regions of MEFV revealed homology to myeloid-specific promoters and to more broadly expressed inflammatory promoter elements. In vitro stimulation of monocytes with the proinflammatory agents interferon (IFN) gamma, tumor necrosis factor, and lipopolysaccharide induced MEFV expression, whereas the antiinflammatory cytokines interleukin (IL) 4, IL-10, and transforming growth factor beta inhibited such expression. Induction by IFN-gamma occurred rapidly and was resistant to cycloheximide. IFN alpha also induced MEFV expression. In granulocytes, MEFV was up-regulated by IFN gamma and the combination of IFN-alpha and colchicine. These results refine understanding of MEFV by placing the gene in the myelomonocytic-specific proinflammatory pathway and identifying it as an IFN-gamma immediate early gene. PMID- 10807794 TI - Perfusion with sickle erythrocytes up-regulates ICAM-1 and VCAM-1 gene expression in cultured human endothelial cells. AB - Sickle cell anemia is characterized by periodic vasoocclusive crises. Increased adhesion of sickle erythrocytes to vascular endothelium is a possible contributing factor to vasoocclusion. This study determined the effect of sickle erythrocyte perfusion at a venous shear stress level (1 dyne/cm(2)) on endothelial cell (EC) monolayers. Sickle erythrocytes up-regulated intercellular adhesion molecule-1 (ICAM-1) gene expression in cultured human endothelial cells. This was accompanied by increased cell surface expression of ICAM-1 and also elevated release of soluble ICAM-1 molecules. Expression of vascular cell adhesion molecule-1 (VCAM-1) messenger RNA (mRNA) was also strikingly elevated in cultured ECs after exposure to sickle cell perfusion, although increases in membrane-bound and soluble VCAM-1 levels were small. The presence of cytokine interleukin-1beta in the perfusion system enhanced the production of ICAM-1 and VCAM-1 mRNA, cell surface expression, and the concentrations of circulating forms. This is the first demonstration that sickle erythrocytes have direct effects on gene regulation in cultured human ECs under well-defined flow environments. The results suggest that perfusion with sickle erythrocytes increases the expression of cell adhesion molecules on ECs and stimulates the release of soluble cell adhesion molecules, which may serve as indicators of injury and/or activation of endothelial cells. The interactions between sickle red blood flow, inflammatory cytokines, and vascular adhesion events may render sickle cell disease patients vulnerable to vasoocclusive crises. PMID- 10807795 TI - Locus control region activity by 5'HS3 requires a functional interaction with beta-globin gene regulatory elements: expression of novel beta/gamma-globin hybrid transgenes. AB - The human beta-globin locus control region (LCR) contains chromatin opening and transcriptional enhancement activities that are important to include in beta globin gene therapy vectors. We previously used single-copy transgenic mice to map chromatin opening activity to the 5'HS3 LCR element. Here, we test novel hybrid globin genes to identify beta-globin gene sequences that functionally interact with 5'HS3. First, we show that an 850-base pair (bp) 5'HS3 element activates high-level beta-globin gene expression in fetal livers of 17 of 17 transgenic mice, including 3 single-copy animals, but fails to reproducibly activate Agamma-globin transgenes. To identify the beta-globin gene sequences required for LCR activity by 5'HS3, we linked the 815-bp beta-globin promoter to Agamma-globin coding sequences (BGT34), together with either the beta-globin intron 2 (BGT35), the beta-globin 3' enhancer (BGT54), or both intron 2 and the 3' enhancer (BGT50). Of these transgenes, only BGT50 reproducibly expresses Agamma-globin RNA (including 7 of 7 single-copy animals, averaging 71% per copy). Modifications to BGT50 show that LCR activity is detected after replacing the beta-globin promoter with the 700-bp Agamma-globin promoter, but is abrogated when an AT-rich region is deleted from beta-globin intron 2. We conclude that LCR activity by 5'HS3 on globin promoters requires the simultaneous presence of beta globin intron 2 sequences and the 260-bp 3' beta-globin enhancer. The BGT50 construct extends the utility of the 5'HS3 element to include erythroid expression of nonadult beta-globin coding sequences in transgenic animals and its ability to express antisickling gamma-globin coding sequences at single copy are ideal characteristics for a gene therapy cassette. PMID- 10807796 TI - A novel mutation in the NADH-cytochrome b5 reductase gene of a Chinese patient with recessive congenital methemoglobinemia. AB - Recessive congenital methemoglobinemia due to nicotinamide adenine dinucleotide (NADH)-cytochrome b5 reductase (b5R) deficiency is classified into 2 clinical types: type 1 (erythrocyte type) and type 2 (generalized type). We found a Chinese family with type 1 recessive congenital methemoglobinemia, the patients from which were diagnosed according to clinical symptoms and b5R enzyme activity in the blood cells. To learn the molecular basis of type 1 recessive congenital methemoglobinemia in this Chinese family, we isolated total RNA from the peripheral leukocytes of the propositus and b5R complementary DNA (cDNA) by reverse transcription- polymerase chain reaction (RT-PCR). The coding region of the b5R cDNA was analyzed by sequencing the cloned PCR products. The results showed that the propositus was homozygous for a G-->A transition at codon 203 in exon 7, changing a cysteine to a tyrosine (Cys203Tyr). To characterize the mutant enzyme, both glutathione S-transferase (GST)-fused wild-type b5R and GST-fused mutant Cys203Tyr b5R were expressed in Escherichia coli and affinity purified. The results showed that the catalytic activity of the enzyme was not much affected by this amino acid substitution, but the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin. These properties of the mutant enzyme would account for the restricted b5R deficiency and mild clinical manifestations of these type 1 patients. The finding of this novel mutation makes codon 203 the only position within the b5R gene at which more than 1 mutation has been found. PMID- 10807797 TI - Herpesvirus saimiri-transformed macaque T cells are tolerated and do not cause lymphoma after autologous reinfusion. AB - Human T cells are transformed in vitro to stable growth after infection with herpesvirus saimiri subgroup C strain C488, and they retain their antigen specific reactivity and other important functional features of mature activated T lymphocytes. The virus persists as nonintegrating episomes in human T cells under restricted viral gene expression and without production of virus particles. This study analyzes the behavior of herpesvirus-transformed autologous T cells after reinfusion into the donor under close-to-human experimental conditions. T cells of 5 macaque monkeys were transformed to stable interleukin-2 dependent growth and were intravenously infused into the respective donor. The animals remained healthy, without occurrence of lymphoma or leukemia for an observation period of more than 1 year. Over several months virus genomes were detectable in peripheral blood cells and in cultured T cells by polymerase chain reaction. In naive control animals, a high-dose intravenous infection rapidly induced pleomorphic peripheral T-cell lymphoma. In contrast, monkeys were protected from lymphoma after challenge infection if they had previously received autologous T-cell transfusions. High levels of antibodies against virus antigens were detectable after challenge infection only. Taken together, herpesvirus-transformed T cells are well tolerated after autologous reinfusion. This may allow us to develop a novel concept for adoptive T-cell mediated immunotherapy. PMID- 10807798 TI - Adoptive immunotherapy in canine mixed chimeras after nonmyeloablative hematopoietic cell transplantation. AB - Development of nontoxic and nonmyeloablative regimens for allogeneic hematopoietic stem-cell transplantation will decrease transplantation-related mortality caused by regimen-related toxic effects. In pursuit of this goal, a dog model of stable mixed hematopoietic chimerism was established in which leukocyte antigen-identical litter mates are given sublethal total-body irradiation (2 Gy) before stem-cell transplantation and immunosuppression with mycophenolate mofetil and cyclosporine afterward. In the current study, we examined whether donor lymphocyte infusion (DLI) could be used as adoptive immunotherapy to convert mixed to complete donor chimerism. First, 8 mixed chimeras were given unmodified DLI between day 36 and day 414 after stem-cell transplantation. After a 10- to 47 week follow-up period, there were no significant changes in the percentage of donor engraftment. Next, we immunized the donor to the minor histocompatibility antigens (mHA) of the recipient by means of repeated skin grafting. Lymphocytes from the mHA-sensitized donor were infused between day 201 and day 651 after transplantation. All 8 recipients of mHA-sensitized DLI had conversion to greater than 98% donor chimerism within 2 to 12 weeks of the infusion. Complications from mHA-sensitized DLI included graft-versus-host disease in 2 dogs and marrow aplasia in 1. These results showed that the low-dose transplant regimen establishes immune tolerance, and mHA-sensitized DLI is required to break tolerance, thereby converting mixed to complete donor chimerism. We propose that mixed chimerism established after nonmyeloablative allogeneic stem-cell transplantation provides a platform for adoptive immunotherapy that has clinical potential in the treatment of patients with malignant diseases. PMID- 10807799 TI - The diagnostic strategy for lung cancer: has determining malignancy or benignity become more accurate? PMID- 10807800 TI - Coronary artery bypass grafting in the elderly: the challenge and the opportunity. PMID- 10807801 TI - The continuing evolution in the management of thoracic aortic dissection. PMID- 10807802 TI - Killip and Forrester classifications: should they be abandoned, kept, reevaluated, or modified? PMID- 10807803 TI - Wet nebulization in acute asthma: the last refrain? PMID- 10807804 TI - Time to move advance care planning beyond advance directives. PMID- 10807805 TI - A multicenter trial with a somatostatin analog (99m)Tc depreotide in the evaluation of solitary pulmonary nodules. AB - OBJECTIVE: The affinity of various malignant neoplasms including small cell and non-small cell lung cancer for peptide analogs of somatostatin has been well documented. Depreotide is such an analog and can be complexed with technetium-99m ((99m)Tc depreotide) for optimal imaging properties. Using this radiopharmaceutical, solitary pulmonary nodules (SPN) were previously evaluated in a successful phase II/III trial. The results of the larger multicenter phase III study using (99m)Tc depreotide to differentiate malignant and benign etiologies in SPN are now presented. METHODS: Patients with SPN 30 years, and no demonstrable radiographic stability for the prior 2 years were studied. All underwent single-photon emission CT (SPECT) with (99m)Tc depreotide and subsequent tissue histologic examination. Three nuclear medicine specialists blinded to histologic findings examined the SPECT images and scored them as positive or negative based on the presence or absence of activity in the radiographic region of the SPN. The final result was determined by the majority score, which was then compared with the histologic result. RESULTS: Of the 114 individuals studied, 88 had a histologic result compatible with malignant neoplasm. (99m)Tc depreotide scintigraphy correctly identified 85 of this group, with three false-negative determinations compared with histology. There were seven false-positive determinations, including six granulomas and one hamartoma. (99m)Tc depreotide scintigraphy correctly excluded malignancy in 19 of 26 patients with benign histologic findings. The sensitivity of this method was 96.6% with a specificity of 73.1%. CONCLUSION: (99m)Tc depreotide scintigraphy is a safe and useful method for the noninvasive evaluation of SPN with a sensitivity and accuracy comparable to that reported for fluorine-18 fluorodeoxyglucose positron emission tomography. PMID- 10807806 TI - Who gets chemotherapy for metastatic lung cancer? AB - STUDY OBJECTIVES: To determine the prevalence and factors associated with chemotherapy use in elderly patients presenting with advanced lung cancer. DESIGN: A retrospective cohort study using administrative data. SETTING AND PATIENTS: We analyzed the medical bills for the 6,308 Medicare patients > 65 years old with diagnosed stage IV non-small cell lung cancer (NSCLC) in the 11 SEER (survival, epidemiology, and end results) regions between 1991 and 1993. The main outcome measure, chemotherapy administration, was identified by the relevant medical billing codes. Patient sociodemographic and disease characteristics were obtained from the SEER database and census data. RESULTS: Almost 22% of patients received chemotherapy at some time for their metastatic NSCLC. As expected, younger patients and those with fewer comorbid conditions were more likely to receive chemotherapy. However, several nonmedical factors, such as nonblack race, higher socioeconomic status, treatment in a teaching hospital, and living in the Seattle/Puget Sound or Los Angeles SEER regions, also significantly increased a patient's likelihood of receiving chemotherapy. CONCLUSION: Compared to previous reports, the prevalence of chemotherapy use for advanced NSCLC appears to be increasing. However, despite uniform health insurance coverage, there is wide variation in the utilization of palliative chemotherapy among Medicare patients, and nonmedical factors are strong predictors of whether a patient receives chemotherapy. While it is impossible to know the appropriate rate of usage, nonmedical factors should only influence a patient's likelihood of receiving treatment if they reflect patient treatment preference. Research to further clarify the costs, benefits, and patient preferences for chemotherapy in this patient population is warranted in order to minimize the effect of nonmedical biases on management decisions. PMID- 10807807 TI - Prognostic value of stage grouping and TNM descriptors in lung cancer. AB - STUDY OBJECTIVES: The International Staging System for Lung Cancer (ISSLC) was revised in 1997. Validation studies are numerous but include only selected surgical patients. This study aims to verify the following: (1) the reliability of the ISSLC in an unselected lung cancer population; (2) the likely improvement in prognostic capability of the new classification; and (3) the possibilities for further improvements. DESIGN: Analysis of a single institution database over a 16 year period from 1983 to 1998. SETTING: Community-based hospital and second referral level institution for a province of 500,000 people. PATIENTS: The study included 1,296 consecutive patients (1,117 men), with pathologically documented lung cancer (46% with squamous cell cancer), staged both clinically (77%) and pathologically (23%), and treated, for the most part, with chemotherapy (52%). INTERVENTIONS: Anthropometric, clinical, and laboratory data were recorded prospectively. Survival analysis was performed by the Kaplan-Meier method and Cox multivariate regression analysis. MEASUREMENT AND RESULTS: The 1997 revised ISSLC classification fit well with the cohort studied. Each stage and substage significantly differed from each other, except for stage IIA. In this stratum, there were only 13 patients. Comparing the 1986 and the 1997 classifications, no substantial differences were observed (log-rank statistics, 295 vs 293, respectively; p < 0.0001). Independent of the classification used, the Cox models were always highly predictive of the outcome. The only way to increase their efficiency was to replace the variable stage with the original TNM descriptors. CONCLUSIONS: Since grouping different TNM subsets into one stage is not really helpful, we might choose to use TNM descriptors in clinical practice and in research. PMID- 10807808 TI - Abnormal flow volume loops in patients with intrathoracic Hodgkin's disease. AB - STUDY OBJECTIVES: To study the incidence of upper airway obstruction, as measured on the flow volume loop (FVL), in patients with bulky mediastinal Hodgkin's disease; to correlate the FVL with CT of the chest; and to follow the changes in the FVL after treatment of the tumor. DESIGN: Retrospective study of pulmonary function tests (PFTs) and chest CTs performed as part of a clinical trial for Hodgkin's disease. SETTING: Memorial Sloan-Kettering Cancer Center, a comprehensive cancer care center. PATIENTS: Twenty-five patients (15 men and 10 women; age range, 20 to 57 years) with bulky mediastinal Hodgkin's disease enrolled in a clinical trial of chemotherapy followed by external beam radiation therapy. MEASUREMENTS AND RESULTS: Fourteen of 25 patients (56%) had an abnormal FVL prior to therapy; after chemotherapy, only 7 of 25 patients (28%) had an abnormal FVL. The abnormal patterns seen were either those typical of fixed obstruction or variable extrathoracic obstruction. No patient had a pattern typical of variable intrathoracic obstruction. On chest CT scan, 16 patients had grade-I tracheal deformity; 6 had grade-II deformity, and 3 had grade-III deformity. All three patients with grade-III deformity had a fixed obstruction pattern, as did three patients with a grade-I pattern. Patients with a fixed pattern on FVL had significant decreases in inspiratory and expiratory flow rates. CONCLUSION: FVL abnormalities suggesting upper airway obstruction occurred in > 50% of patients with bulky mediastinal Hodgkin's disease. A fixed pattern of obstruction was associated with the lower flow rates and severe tracheal distortion on CT; these patients may warrant special attention prior to general anesthesia or invasive procedures. Asymptomatic patients with abnormal FVLs but normal tracheal profiles need not undergo extensive evaluation. No patients showed the expected pattern typical of intrathoracic obstruction, but rather the major effect was on the inspiratory loop. The authors speculate on the mechanism for this unexpected finding. PMID- 10807809 TI - Coronary artery bypass grafting in the elderly. AB - BACKGROUND AND METHODS: The incidence of coronary artery bypass grafting (CABG) in elderly patients has been increasing. We retrospectively analyzed the results of CABG performed at Shin-Tokyo Hospital between January 1, 1991, and December 31, 1998. Preoperative, perioperative, and follow-up data of patients > or = 75 years old (group E, n = 190) were collected, and compared with those of patients < 75 years old (group Y, n = 1,380). RESULTS: Female gender, emergent CABG, preoperative balloon pumping use, cardiogenic shock, hypertension, and preoperative cerebral vascular accident were significantly more frequent in group E (p < 0.05). CABG was completed without any significant differences, except for less frequent use of the bilateral internal mammary artery (p < 0.01), more frequent use of the saphenous vein (p < 0.005), and a greater incidence of blood transfusion in group E (p < 0.0001). The postoperative course required longer intubation, ICU stay, and postoperative hospital stay in group E (p < 0.001), and was more frequently associated with major complication (p < 0.0001) and in hospital death (p < 0.05). During the mean follow-up of 2.7 years (maximum 6.9 years), the actuarial 5-year survival of groups E and Y were 84.3% and 92.5% (p < 0.01), respectively, excluding in-hospital mortality. The actuarial 5-year cardiac event-free rates were 79.9% in group E and 79.7% in group Y, showing no significant difference. CONCLUSIONS: CABG in the elderly carries certain surgical risks. However, the long-term cardiac event-free rate after CABG in the elderly was almost the same as that of younger patients. Inferior long-term survival in the elderly was most likely due to the biological nature of aging. PMID- 10807810 TI - Epidemiology and clinicopathology of aortic dissection. AB - STUDY OBJECTIVES: To determine the incidence and mortality as well as to analyze the clinical and pathologic changes of aortic dissection. DESIGN AND SETTING: A population-based longitudinal study over 27 years on a study population of 106,500, including 66 hospitalized and 18 nonhospitalized consecutively observed patients. MEASURES: Analysis of data from the medical, surgical, and autopsy records of patients with aortic dissection. RESULTS: Altogether, 86 cases of aortic dissection were found in 84 patients, corresponding to a 2.9/100,000/yr incidence. Sixty-six of the 84 patients (79%) were admitted to the hospital, and 18 patients (21%) died before admission. Their ages ranged from 36 to 97 years, with a mean of 65. 7 years. The male/female ratio was 1.55 to 1. A total of 22.7% of the hospitalized patients died within the first 6 h, 33.3% within 12 h, 50% within 24 h, and 68.2% within the first 2 days after admission. Six patients were operated on, with a perioperative mortality of two of six patients and a 5-year survival of two of six patients. All patients who were not operated on died. Pain was the most frequent initial symptom. Every patient had some kind of cardiovascular and respiratory sign. Neurologic symptoms occurred in 28 of 66 patients (42%). Five patients presented with clinical pictures of acute abdomen and two with acute renal failure. Trunk arteries were affected in 33 of the 80 autopsied cases (41%), and rupture of aorta was seen in 69 cases (86%). In five cases, spontaneous healing of dissection was also found. The ratio of proximal/distal dissections was 5.1 to 1. All 18 prehospital cases were acute. Fifty-nine cases (89.4%) were acute at admission, and 7 cases (10.6%) were chronic dissections. Hypertension and advanced age were the major predisposing factors. CONCLUSION: Aortic dissection was the initial clinical impression in only 13 of the 84 patients (15%). Thus, 85% of the patients did not receive immediate appropriate medical treatment. For this reason, these late-recognized and/or unrecognized cases may be regarded as an untreated or symptomatically treated group, whose course may resemble the natural course of aortic dissection. PMID- 10807811 TI - Pneumothorax: experience with 1,199 patients. AB - OBJECTIVE: To study the outcome of pneumothorax managed in a university affiliated metropolitan medical center. DESIGN: A retrospective review. SETTING: Busy metropolitan medical center. PATIENTS AND METHODS: Records of 1,199 patients with pneumothorax were reviewed and analyzed. RESULTS: Primary spontaneous pneumothorax occurred in 218 patients, secondary spontaneous pneumothorax occurred in 505, traumatic in 403, and iatrogenic in 73. Ninety-six patients with small pneumothorax (8%) were managed by observation, and 1,103 patients (92%) were managed by tube thoracostomy. Drainage of the pleural cavity was continued for 1 to 7 days in 893 patients (81%), 8 to 10 days in 176 patients (16%), and > 10 days in 34 patients (3%). Drainage for > 10 days was classified as persistent pneumothorax. In these 34 patients and in 132 others with a second ipsilateral recurrence (a total of 166 patients), direct pleuroscopy was performed. The pleuroscopy findings and further management are outlined in the algorithm. CONCLUSIONS: Pneumothorax is a common condition affecting all age groups. If the volume of the pneumothorax is > 20% of the pleural space, pleural drainage is indicated. For management of persistent or recurrent pneumothorax, the use of pleuroscopy (direct or video-assisted) is of great value and should be part of routine management. PMID- 10807812 TI - Hemodynamic classification in acute myocardial infarction. AB - OBJECTIVE: Current mortality (M(1)) in hemodynamic subgroups of patients with acute myocardial infarction (AMI) was compared to that observed 30 years ago (M(0)), when hemodynamic classification was established. The prognostic value of oxyhemodynamic indexes in predicting M(1) for patients receiving right heart catheterization (RHC) was investigated. PATIENTS AND METHODS: We assigned 393 patients with AMI (mean age, 72 +/- 10 years) to four Killip categories. A fiberoptic reflectance catheter was inserted in the pulmonary artery (PA) in 136 patients. Cardiac index (CI), PA wedge pressure (PWP), PA mixed venous blood oxygen saturation (SvO(2)), oxygen extraction ratio (O(2)ER), and normalized CI (NCI; CI/O(2)ER) were measured. Catheterized patients were classified into four Forrester groups, and M(1) and M(0) were compared. Survivors (group S) were compared to nonsurvivors (group NS), and the prognostic value of oxyhemodynamic parameters in predicting M(1) was estimated. RESULTS: A significant decline in total mortality was observed (M(1) of 8% vs M(0) of 34%; p < 0.0001). In catheterized patients, total M(1) was also decreased (M(1) of 15% vs M(0) of 26%; p < 0.05). Compared with group S, group NS had lower (mean +/- SD) CI (1.8 +/- 0.4 L/min/m(2) vs 2.4 +/- 0.6 L/min/m(2); p < 0.01), SvO(2) (46.1 +/- 10.6% vs 59.9 +/- 10.0%; p < 0.01), NCI (4.2 +/- 1.4 vs 7.4 +/- 4.1 L/min/m(2); p < 0.01), and higher PWP (22.7 +/- 6.8 mm Hg vs 14. 4 +/- 4.7 mm Hg; p < 0.01). NCI presented the best sensitivity (81%), specificity (78%), and predictive value (40%), in predicting M(1). CONCLUSIONS: The historical AMI hemodynamic classification has lost its semiquantitative value, since mortality has decreased. RHC does not compromise the outcome. NCI has a high prognostic value in predicting early mortality. PMID- 10807813 TI - Habitual activities and peak aerobic capacity in patients with asymptomatic and symptomatic left ventricular dysfunction. AB - BACKGROUND: A reduced level of daily activities is thought to be an important determinant of aerobic exercise intolerance in patients with chronic heart failure chronic heart failure; however, few data exist about the relationship between habitual physical activity level and peak aerobic capacity in patients at different clinical stages of left ventricular dysfunction. STUDY OBJECTIVES: The purposes of this study were as follow: (1) to validate a simple interviewer administered scoring system for evaluation of habitual physical activity level of patients with chronic heart failure and asymptomatic left ventricular dysfunction (ALVD); (2) to determine the relationship between habitual physical activity level and peak aerobic capacity in chronic heart failure and ALVD patients; and (3) to compare habitual activity levels among different New York Heart Association (NYHA) classes in these populations. SETTING: Cardiology division at a tertiary-care hospital. STUDY POPULATION: We studied 167 consecutive patients with chronic heart failure (NYHA class I to III), 40 patients with ALVD, and 52 healthy subjects (HS). MEASUREMENTS AND RESULTS: Habitual physical activity level was evaluated by means of an interview-based activity scoring system considering leisure time and occupational activities and also recent deconditioning events (eg, hospital admissions); a final activity score (AS) ranging from 0.8 to 5 was obtained. All patients and HS performed a symptom-limited cardiopulmonary exercise test up to a respiratory exchange ratio of > or = 1.1. AS was an independent predictor of peak oxygen consumption (VO(2)) in all groups, with a significantly higher VO(2) vs AS relationship slope in the ALVD and HS groups than in the chronic heart failure group. Moreover, AS was found to be significantly lower in chronic heart failure than in ALVD patients and HS (1.6 +/ 0.6 vs 2.2 +/- 0.7 vs 3.5 +/- 1.1, respectively; p < 0.0001), as was peak VO(2) (14.7 +/- 3.7 mL/kg/min vs 20 +/- 4 mL/kg/min vs 33.1 +/- 10 mL/kg/min, respectively; p < 0.0001), but the latter differences were canceled after adjusting for AS values. Significant AS and peak VO(2) reductions were observed in chronic heart failure patients with NYHA class progression from I to III. CONCLUSIONS: Habitual physical activity level is progressively decreased with worsening of heart failure symptoms and is related to peak aerobic capacity in both chronic heart failure and ALVD patients. However, this relationship appears to be weak in patients with chronic heart failure, whereas daily activity is a strong independent predictor of peak aerobic capacity both in ALVD patients and HS. This may be related to the intervention of factors other than skeletal muscle deconditioning in the exercise pathophysiology of chronic heart failure patients. PMID- 10807814 TI - The effect of coronary vasospasm on the direction of ST-segment deviation in patients with both hypertrophic cardiomyopathy and vasospastic angina. AB - BACKGROUND: There has been no report of ECG changes during anginal attacks in patients with coexistent hypertrophic cardiomyopathy (HCM) and vasospastic angina. STUDY OBJECTIVES: To elucidate the change in ST-segment during anginal attacks in patients with coexistent HCM and vasospastic angina (the HCM group) in comparison with that of patients with vasospastic angina and no left ventricular hypertrophy (the non-HCM group). DESIGN: Retrospective study. PATIENTS: Twelve patients in the HCM group, and 28 patients in the non-HCM group. MEASUREMENTS: The direction of ST segment shift, either ST-segment elevation or depression, on the ECGs recorded during vasospastic anginal attacks with severe vasoconstriction in the epicardial coronary artery after intracoronary injection of acetylcholine. RESULTS: Age, male gender, and distribution of coronary arteries in which the vasospasm occurred were similar between the two groups. Collateral circulation to the affected arteries was absent in all the study patients. The prevalence of anginal attacks associated with ST-segment elevation was 2.7 times higher in the non-HCM group than in the HCM group (51. 5% [17 of 33 attacks] vs 18.8% [3 of 16 attacks], respectively; p = 0.03). CONCLUSIONS: In the HCM group, myocardial ischemia associated with a transmural injury pattern seen on the ECG, which is represented as ST-segment elevation, seldom develops during vasospastic anginal attacks because of marked left ventricular hypertrophy. PMID- 10807815 TI - Is nebulized aerosol treatment necessary in the pediatric emergency department? AB - BACKGROUND: Infants and small children admitted to the pediatric emergency department (PED) with acute wheezing episodes (AWE) are currently treated with nebulized wet aerosol (NWA). OBJECTIVE: To determine the efficacy of MDI with Nebuchamber (Astra AB; Lund, Sweden), a nonelectrostatic spacer device (NESD), as compared to NWA in the treatment of an unselected population of babies and small children with AWE. DESIGN: Randomized, double-blind, placebo-controlled trial. Forty-two children referred to the PED (median age +/- SD, 16 +/- 15 months) with AWE received either placebo MDI through a NESD (four puffs) and salbutamol 0.5 mL (2.5 mg) as a NWA (group I, n = 19), or salbutamol MDI and 0.5 mL of saline solution administered in the same manner as above (group II, n = 23). This treatment was repeated three times every 20 min. RESULTS: The respiratory rates (RRs) at baseline were as follows: group I, 45 +/- 11.2 breaths/min; and group II, 52.3 +/- 11.3 breaths/min (p = not significant [NS]). After the first, second, and third interventions, the percent fall from baseline of the RR were as follows: group I, 8.9, 13.1, and 17.9%, respectively; group II, 8. 6, 14.6, and 18.6%, respectively. There was no significant difference at any time in the results between the two groups. The clinical scores (CSs) at baseline were as follows: group I, 6.6 +/- 1.3; group II, 6.8 +/- 1.49 (p = NS). After the first, second, and third interventions, the percent fall from baseline of the CS were as follows: group I, 9.1, 17.9, and 23.2%, respectively; group II, 8. 6, 18.9, and 24.7%, respectively. These results, also, did not differ significantly at any time between the two groups. Hospitalization rate and side effects did not differ between the two groups. CONCLUSIONS: We conclude that even in the group of unselected very young children (mean age < 2 years) with AWE, the use of MDI with NESD is at least as effective as the use of NWA. As opposed to data from an adult population, no plateau was reached in the dose-response curve using the above doses over time. PMID- 10807817 TI - The protective effect of salbutamol inhaled using different devices on methacholine bronchoconstriction. AB - STUDY OBJECTIVE: To determine the protective effect of salbutamol, 100 microg, inhaled by different devices (pressurized metered-dose inhaler [pMDI; Ventolin; GlaxoWellcome; Greenford, UK], pMDI + spacer [Volumatic; GlaxoWellcome], or breath-activated pMDI [Autohaler; 3M Pharmaceuticals; St. Paul, MN]) on bronchoconstriction induced by methacholine. DESIGN: Randomized, double-blind, cross-over, placebo-controlled study. PATIENTS: Eighteen subjects with stable, moderate asthma, asymptomatic, receiving regular treatment with salmeterol, 50 microg bid, and inhaled beclomethasone dipropionate, 250 microg bid, in the last 6 months, with high hyperreactivity to methacholine (baseline provocative dose of methacholine causing a 20% fall in FEV(1) [PD(20)] geometric mean [GM], 0.071 mg). Subjects were classified into two groups: subjects with incorrect (n = 5) pMDI inhalation technique, and subjects with correct (n = 13) inhalation technique. METHODS AND MEASUREMENTS: After cessation of therapy for 3 days, all subjects underwent four methacholine challenge tests, each test 1 week apart, each time 15 min after inhalation of salbutamol, 100 microg (via pMDI, pMDI + spacer, or Autohaler), or placebo. The protective effect on methacholine challenge test was evaluated as the change in the PD(20), and expressed in terms of doubling doses of methacholine in comparison with placebo treatment. RESULTS: The PD(20) was significantly higher after salbutamol inhalation than after placebo inhalation, but no significant difference was observed among the three different inhalation techniques. Only when salbutamol was inhaled via pMDI + spacer, PD(20) was slightly but not significantly higher (pMDI GM, 0.454 mg; pMDI + spacer GM, 0.559 mg; and Autohaler GM, 0.372 mg; not significant [NS]) than other inhalation techniques. Similar results (mean +/-SEM) were obtained with doubling doses of methacholine (pMDI, 2 +/- 0.47; pMDI + spacer, 3 +/- 0.35; and Autohaler, 2.4 +/- 0.40; NS). No significant difference was found among techniques when subjects with correct or incorrect inhalation technique were separately considered. CONCLUSIONS: Our data show that the protective effect of salbutamol, 100 microg, on methacholine-induced bronchoconstriction is not affected by the different inhalation techniques, although inhalation via pMDI + spacer tends to improve the bronchoprotective ability of salbutamol. These data confirm the clinical efficacy of salbutamol, whatever the device, and the patient's inhalation technique. PMID- 10807816 TI - Salmeterol administration by metered-dose inhaler alone vs metered-dose inhaler plus valved holding chamber. AB - STUDY OBJECTIVE: To determine whether a spacer device designed as a valved holding chamber with a flow signal increases the efficacy of the long-acting beta(2)-agonist, salmeterol, in patients who use incorrect technique with metered dose inhaler (MDI) alone. DESIGN: Double-blind, randomized, placebo-controlled study. SETTING: University hospital outpatient rooms. PATIENTS: Twenty adult outpatients with stable persistent asthma, receiving a daily anti-inflammatory drug. INTERVENTIONS: Patients were randomized to either salmeterol MDI (incorrect use: 1 s after actuating MDI, inhale rapidly) and placebo plus spacer (correct use: inhale slowly as MDI is actuated, continue to inhale slowly and deeply) or placebo MDI (incorrect use) and salmeterol plus spacer (correct use). The following week, patients received the opposite treatment. The dose was two puffs from each device on each treatment day; each puff was separated by 1 min. MEASUREMENTS AND RESULTS: After baseline peak expiratory flow (PEF), salmeterol was administered and serial PEF determined (0.5, 1, 2, 3, 4, 6, 8, 10, and 12 h). Administration of salmeterol MDI plus spacer resulted in significantly greater increases in PEF from baseline vs MDI at 4 h (44 L/min vs 10 L/min; p < 0.01) and 6 h (49 L/min vs 24 L/min; p < 0.05). Both methods of administration were equally well tolerated. CONCLUSION: We conclude that patients who have poor timing and rapid inhalation with salmeterol MDI alone will have greater increases in PEF at 4 h and 6 h and no additional side effects if the dose is administered with a valved holding chamber that is used correctly. Further study is needed regarding other errors in MDI technique with salmeterol. PMID- 10807818 TI - Prevalence of cocaine use and its impact on asthma exacerbation in an urban population. AB - OBJECTIVES: To assess the prevalence of cocaine use, and its impact on severity of presentation, among adults presenting to the emergency department (ED) with asthma. A secondary aim was to assess the use of various asthma treatment modalities, with reference to the 1997 National Asthma Education and Prevention Program (NAEPP) guidelines. METHODS: All adults aged 18 to 55 years who presented to the ED of this institution with an asthma attack, were approached about participating in the study, which required giving informed consent, answering a facilitated questionnaire, and giving a urine sample for drug screening. RESULTS: Patients were enrolled during a 7-month period. A total of 163 patients were approached to enter the study; 116 patients consented to participate in the study, with 103 submitting complete urine samples. Thirty-seven patients refused to participate, and 10 were excluded. Sixty-eight percent of the patients were women, with a mean age of 33 years. African-Americans made up 89% of the total group. Thirty-five percent were cigarette smokers. Urine cocaine tests were positive in 13 of 103 (13%); 6 of 103 (5.8%) were positive for opiates. In the cocaine-positive group, 5 of 13 patients (38%) were admitted to the hospital, including two patients requiring intubation and mechanical ventilation. Of the total group, 23 of 103 patients (22%) were admitted, and 5 of those 23 admitted patients (22%) were cocaine-positive. Length of stay was significantly longer (5 vs 2.5 days, p < 0.05) in the cocaine-positive admitted patients. Forty-six percent of all patients reported using inhaled corticosteroids (ICS), with 39% of admitted patients using them. Thirty-two percent of all patients had obtained three or more refills of their beta(2)-agonist inhaler in the previous month. CONCLUSIONS: The prevalence of cocaine use may be much higher than the 13% shown in this study, because of patients' refusal to participate in the study. Second, the severity of exacerbation appears to be worse in the cocaine-positive group. Finally, the majority of patients presenting did not use ICS in accordance with the NAEPP guidelines. PMID- 10807819 TI - Impact of airway lability, atopy, and tobacco smoking on the development of asthma-like symptoms in asymptomatic teenagers. AB - AIM: To investigate the impact of airway lability, atopy, and tobacco smoking on the development of asthma-like symptoms in asymptomatic subjects. METHODS: In this prospective, community-based study, 271 asymptomatic adolescents with an average age at inclusion of 13.9 years were followed for 6.4 years. Airway lability was assessed at baseline by three tests, including exercise challenge, airway provocation with methacholine, and monitoring of peak expiratory flow. Atopy was defined by one or more positive reactions (> or = 3-mm weal) to 10 common aeroallergens by skin prick testing. The influence of airway lability, atopy, and smoking on the development of asthma-like symptoms was assessed by logistic regression. RESULTS: During the 6-year study period, 68 of the previously asymptomatic teenagers (25%) developed asthma-like symptoms. Among those, 50% reported cough only, 29% reported wheezing only, and 21% reported both wheezing and coughing. Hyperresponsiveness to methacholine (odds ratio [OR], 3.5; 95% confidence interval [CI], 1.1 to 11.6), smoking (OR, 2.1; 95% CI, 1. 2 to 3.8), and atopy (OR, 3.5; 95% CI, 1.8 to 6.8) each contributed independently to explain symptom development (wheezing and cough together). Girls, but not boys, with airway lability were less likely to take up smoking, compared with subjects of that set with no airway lability (32% vs 51%; p < 0.05). No effect of airway lability on the likelihood of giving up smoking could be demonstrated, nor did the presence of atopy have any significant impact on smoking behavior. CONCLUSION: Hyperresponsiveness to methacholine, atopy, and smoking were independent risk factors for the development of asthma-like symptoms during adolescence. The presence of airway lability may prevent girls from taking up smoking. PMID- 10807820 TI - Racial differences in physiologic parameters related to asthma among middle-class children. AB - BACKGROUND: Asthma morbidity and mortality are higher in the United States for African-American (AA) children when compared to European-American (EA) children. STUDY OBJECTIVES: To explore racial differences in physiologic factors associated with pediatric asthma severity. DESIGN: Cross-sectional. METHODS: We analyzed data from two groups of children in suburban Detroit, one of which contains non urban, middle-class AA children, a group not usually included in childhood asthma studies. All children were 6 to 8 years of age. Clinical evaluations included medical history, physical examination, skin testing, spirometry, and methacholine challenge. RESULTS: The study population (n = 569) was 14% African American, 51% of the participants were male, and the mean age was 6.8 +/- 0.4 years. Socioeconomic status (parental education) was similar overall by race, although some strata-specific differences were observed. The prevalence of physician diagnosed asthma was 10% for both AA and EA groups. AA children were more reactive to methacholine than EA children (42% vs 22%, respectively; p = 0.001), and had significantly higher total IgE than EA children (geometric mean, 60. 6 vs 27.5 IU/mL; p = 0.001). Serum IgE was related to methacholine reactivity in EA children (p = 0.001), but not AA children (p = 0. 73). These differences remained after adjustment for gender, age, parental education, parental smoking, and maternal smoking during pregnancy. CONCLUSIONS: Our data support previous reports of racial differences in lung volume, airway responsiveness, and serum IgE concentrations. We found a racial difference in the relationship between total serum IgE and airway responsiveness that is unreported elsewhere. Overall, our results suggest that AA children may be predisposed to asthma. PMID- 10807821 TI - Antibiotics are associated with lower relapse rates in outpatients with acute exacerbations of COPD. AB - BACKGROUND: COPD is a complex disease with exacerbations characterized by worsening of symptoms resulting in deteriorating lung function. STUDY OBJECTIVE: To assess predictive factors of relapse for patients with acute exacerbations of COPD (AECB). DESIGN: Retrospective cohort analysis of visits for AECB. SETTING: Veterans Affairs Medical Center. PATIENTS: Three hundred sixty-two visits (173 patients) with documented COPD treated as outpatients for AECB. MEASUREMENTS: Severity of underlying COPD, severity of AECB, comorbid conditions, therapy, and relapse rates (return visit within 14 days with persistent or worsening symptoms). RESULTS: Each visit was analyzed individually (referred to as a patient-visit). One group received antibiotics (270 patient-visits), and the second group (92 patient-visits) did not. Both groups had similar demographics and severity of underlying COPD. The overall relapse rate was 22%. The majority of patient-visits (95%) with severe symptoms at presentation were prescribed antibiotics vs only 40% of those with mild symptoms. Twenty-nine of 92 patient visits (32%) were followed by relapse in the group that was not given antibiotics, whereas only 50 of 270 (19%) treated with antibiotics relapsed (p < 0.001). Those treated with amoxicillin had an even higher relapse rate (20 of 37 patient-visits, or 54%) than those who did not receive antibiotics (p = 0.006). CONCLUSIONS: Relapse from AECB was not related to the severity of underlying disease or to the severity of the acute exacerbation. Patients treated with antibiotics had significantly lower relapse rates than those who did not receive antibiotics. However, the specific choice of antibiotic is important because those treated with amoxicillin had the highest relapse rates of all groups. PMID- 10807822 TI - Tumor necrosis factor gene complex in COPD and disseminated bronchiectasis. AB - BACKGROUND: Tumor necrosis factor (TNF) is a potent proinflammatory cytokine with increased levels in the sputum of COPD subjects. Two biallelic TNF gene complex polymorphisms have been described: LtalphaNcoI, in the first intron of the lymphotoxin alpha (previously referred to as TNF-beta) gene, and TNF-308, in the promoter region of the TNF-alpha gene. Higher levels of TNF production are associated with allele 1 of LtalphaNcoI (LtalphaNcoI*1) and with allele 2 of TNF 308 (TNF-308*2). STUDY OBJECTIVES: To study the frequencies of the two TNF gene complex polymorphisms in patients with COPD and bronchiectasis. DESIGN: Association study. SUBJECTS AND METHODS: We studied the frequencies of these polymorphisms in 66 subjects with COPD and in 23 subjects with disseminated bronchiectasis and compared them to the frequencies in 98 healthy control subjects and 45 subjects with nonobstructive pulmonary disease. Genomic DNA samples were extracted, and TNF-alpha and LtalphaNcoI polymorphisms were detected after polymerase chain reaction by restriction digestion. RESULTS: We found the following frequencies: the TNF-308*2 allele was detected in 11% of COPD individuals, 15% of bronchiectasis patients, 10% of healthy control subjects, and 18% of subjects with nonobstructive pulmonary disease. The LtalphaNcoI*1 allele was detected in 28% of COPD individuals, 30% of bronchiectasis patients, 29% of healthy control subjects, and 29% of subjects with nonobstructive pulmonary disease. We found evidence of linkage disequilibrium between the two loci (Delta = 0.068). CONCLUSIONS: We conclude that the TNF gene complex, at least in Caucasoid individuals and for the considered polymorphisms, does not seem to play a major role as genetic risk factor in COPD and bronchiectasis. PMID- 10807823 TI - Quantitating physical activity in COPD using a triaxial accelerometer. AB - STUDY OBJECTIVE: To determine the reliability, validity, and stability of a triaxial accelerometer for walking and daily activity measurement in a COPD sample. DESIGN: Cross-sectional, correlational, descriptive design. SETTING: Outpatient pulmonary rehabilitation program in a university-affiliated Veterans Affairs medical center. PARTICIPANTS: Forty-seven outpatients (44 men and 3 women) with stable COPD (FEV(1), 37% predicted; SD, 16%) prior to entry into a pulmonary rehabilitation program. MEASUREMENTS AND RESULTS: Test-retest reliability of a triaxial movement sensor (Tritrac R3D Research Ergometer; Professional Products; Madison, WI) was evaluated in 35 of the 47 subjects during three standardized 6-min walks (intraclass correlation coefficient [rICC] = 0.84). Pearson correlations evaluated accelerometer concurrent validity as a measure of walking (in vector magnitude units), compared to walking distance in all 47 subjects during three sequential 6-min walks (0. 84, 0.85, and 0.95, respectively; p < 0.001). The validity of the accelerometer as a measure of daily activity over 3 full days at home was evaluated in all subjects using Pearson correlations with other indicators of functional capacity. The accelerometer correlated with exercise capacity (maximal 6-min walk, r = 0.74; p < 0.001); level of obstructive disease (FEV(1) percent predicted, r = 0.62; p < 0.001); dyspnea (Functional Status and Dyspnea Questionnaire, dyspnea over the past 30 days, r = - 0.29; p < 0.05); and activity self-efficacy (Activity Self-Efficacy Questionnaire, r = 0.43; p < 0.01); but not with self-report of daily activity (Modified Activity Recall Questionnaire, r = 0.14; not significant). Stability of the accelerometer to measure 3 full days of activity at home was determined by an rICC of 0.69. CONCLUSIONS: This study provides preliminary data suggesting that a triaxial movement sensor is a reliable, valid, and stable measure of walking and daily physical activity in COPD patients. It has the potential to provide more precise measurement of everyday physical functioning in this population than self report measures currently in use, and measures an important dimension of functional status not previously well-described. PMID- 10807824 TI - Implementation of admission decision support for community-acquired pneumonia. AB - STUDY OBJECTIVES: Considerable variation exists in hospital admission rates for patients with community-acquired pneumonia. Logic to determine need for admission has been proposed by several authors. We compared Intermountain Health Care pneumonia guideline recommendations for inpatient vs outpatient care with actual physician decision making and clinical outcomes before vs after implementation. A secondary objective was to determine whether the pneumonia severity index predicts need for admission in this population. DESIGN: Prospective study after implementation vs historic controls. SETTING: Four ambulatory, urgent-care facilities. PATIENTS: Four hundred sixty-three immunocompetent adults with radiographically confirmed community-acquired pneumonia. INTERVENTION: A pneumonia practice guideline including decision support logic was implemented for a 12-month period. MEASUREMENTS AND RESULTS: After implementation, physicians used the pneumonia guideline form in 90% of cases. The percentage of patients admitted within 30 days decreased from 13.6% to 6.4% (p = 0.01). Only five patients before (2.5%) and three patients after (1.1%, p = 0.3) guideline implementation required subsequent hospital admission within 30 days after initial outpatient treatment. Only two deaths occurred in the study cohort, both outpatients before implementation. The positive predictive value was 14.4%, and the negative predictive value for admission was 98.8% after guideline implementation. Guideline recommendation for admission was more likely to be followed in patients with more risk factors and hypoxemia. CONCLUSIONS: Decreased admission rate was observed after implementation of admission decision support in combination with specific recommendations for outpatient antibiotic therapy. Favorable outpatient outcomes suggest that implementation of decision support was safe. PMID- 10807825 TI - Process of care performance, patient characteristics, and outcomes in elderly patients hospitalized with community-acquired or nursing home-acquired pneumonia. AB - STUDY OBJECTIVES: To compare process of care performance, patient characteristics, and outcomes in a contemporary cohort of elderly (> or = 65 years) patients hospitalized with community-acquired pneumonia (CAP) or with nursing home-acquired pneumonia (NHAP). DESIGN: State-wide retrospective cohort study. SETTING: Thirty-four acute-care hospitals in Connecticut. PATIENTS: Elderly Medicare patients hospitalized in 1995-1996 with CAP (1,131) or with NHAP (528). MEASUREMENTS: Antibiotic administration within 8 h of hospital arrival, blood culture collection within 24 h of hospital arrival, oxygenation assessment within 24 h of hospital arrival, demographic and clinical characteristics, in hospital complications, mortality, and length of stay. RESULTS: Process of care performance rates for patients with CAP and NHAP were equivalent for antibiotic administration within 8 h of hospital arrival (76.8% vs 76.3%, respectively; p = 0.82), blood culture collection within 24 h of hospital arrival (78.1% vs 81.1%, respectively; p = 0.31), and oxygenation assessment within 24 h of hospital arrival (94.7% vs 95. 3%, respectively; p = 0.70). Patients with CAP were younger than those with NHAP (median age, 80 vs 84 years, respectively; p < 0. 001), had less cerebrovascular disease (16.8% vs 34.7%, respectively; p < or = 0.001), and lower mortality risk scores at hospital presentation (median, 100 vs 137, respectively; p < or = 0. 001) than patients with NHAP. The median length of stay was equivalent (7 days), but the in-hospital mortality rate was lower in patients with CAP than in patients with NHAP (8.0% vs 18.6%, respectively; p < or = 0.001). CONCLUSION: Initial hospital processes of care are performed at the same rate in patients hospitalized with CAP or NHAP. However, patients with CAP are younger, are less acutely and chronically ill, and have lower in-hospital mortality rates than patients with NHAP. PMID- 10807826 TI - Acute chest syndrome in adults with sickle cell disease. AB - STUDY OBJECTIVES: Acute chest syndrome (ACS) is a frequent and potentially severe pulmonary illness in sickle cell disease (SCD). The aim of the study was to report the clinical features and outcome of consecutive ACS episodes in adult patients in a French SCD center. All patients were treated according to an uniform therapeutic protocol applying transfusion only in the more severe clinical form of ACS. RESULTS: There were 107 consecutive episodes in 77 adult patients (mean age, 29 +/- 7 years; 78% hemoglobin [Hb] SS; 14% Hb SC; and 8% Hb Sbeta + thalassemia) over a 6-year period. Seventy-eight percent of our patients had an associated vaso-occlusive crisis that preceded the chest signs in half of the cases. Comparison between acute and baseline levels showed a statistically significant difference in Hb levels (drop of 1.6 to 2. 25 g/dL depending on Hb genotype), WBC count (increase of 9.2 +/- 8. 3 x 10(9)/L); platelet count (increase of 67 +/- 209 x 10(9)/L); and lactate dehydrogenase values (increase of 358 +/- 775 IU/L) in ACS patients. Hypercapnia was detected in 42% of patients without sign of narcotic abuse. We identified a high percentage of alveolar macrophages containing fat droplets in 31 of 43 (77%) patients who underwent BAL. Bacterial culture findings were almost always negative, but were performed after starting antibiotic therapy that was administered in 96 episodes. Transfusion was required in 50 of 107 ACS events (47%). Five patients died, and all were transfused. CONCLUSIONS: These results confirm that fat embolism is probably a frequent mechanism of ACS in adult patients. However, fat embolism was not associated with a more severe clinical course, suggesting that bronchoscopy and BAL have little impact on the management of these patients. Restricting transfusion to the most severe ACS cases does not seem to increase the mortality rate. PMID- 10807827 TI - Urinary leukotriene E(4) levels are not increased prior to high-altitude pulmonary edema. AB - STUDY OBJECTIVE: To examine whether increased urinary cysteinyl-leukotriene E(4) (LTE(4)) excretion, which has been found to be elevated in patients presenting with high-altitude pulmonary edema (HAPE), precedes edema formation. DESIGN: Prospective studies in a total of 12 subjects with susceptibility to HAPE. SETTING: In a chamber study, seven subjects susceptible to HAPE and five nonsusceptible control subjects were exposed for 24 h to an altitude of 450 m (control day), and exposed for 20 h to 4,000 m after slow decompression over 4 h. In a field study, prospective measurements at low and high altitude were performed in five subjects developing HAPE at 4,559 m. PARTICIPANTS: Mountaineers with a radiographically documented history of HAPE and control subjects who did not develop HAPE with identical high-altitude exposure. INTERVENTIONS: 24-h urine collections. MEASUREMENTS AND RESULTS: In the hypobaric chamber, none of the subjects developed HAPE. The 24-h urinary LTE(4) did not differ between HAPE susceptible and control subjects, nor between hypoxia and normoxic control day. In the field study, urinary LTE(4) was not increased in subjects with HAPE compared to values obtained prior to HAPE at high altitude and during 2 control days at low altitude. CONCLUSIONS: These data do not provide evidence that cysteinyl-leukotriene-mediated inflammatory response is associated with HAPE susceptibility or the development of HAPE within the context of our studies. PMID- 10807828 TI - Pulmonary arteriovenous malformations. AB - OBJECTIVE: Helical CT scan (HCT), a noninvasive method, can detect pulmonary arteriovenous malformations (PAVMs). Its sensitivity is superior to that of global digitalized angiography, but patients receive a significant dose of radiation during diagnostic HCT. We compared HCT to contrast-enhanced pulmonary magnetic resonance angiography (CEMRA), a new noninvasive radiation-free method, in the diagnosis of PAVMs. PATIENTS AND METHODS: Five consecutive patients with PAVMs underwent HCT, CEMRA, and pulmonary artery digital subtraction angiography (PADSA). CEMRA was performed during the pulmonary arterial phase of an IV bolus of gadolinium. PADSA was performed during the embolization procedure. All images were examined for PAVMs. The site and size of aneurysms were specified, as well as the diameter of the vascular pedicles. RESULTS: Thirty PAVMs were detected by CEMRA and 38 by HCT. All 20 PAVMs at least 5 mm in diameter and 10 of the 18 PAVMs < 5 mm in diameter identified on HCT were also identified by CEMRA. Whatever the site, all PAVMs with a feeding artery diameter of at least 3 mm (ie, PAVMs with clinical consequences) were detected by CEMRA. No false-positive results were obtained with CEMRA. CEMRA therefore had a sensitivity of 78% and a specificity of 100%. CONCLUSIONS: CEMRA, a nonionizing and noninvasive procedure, has high sensitivity and specificity for the diagnosis of clinically relevant PAVMs. PMID- 10807829 TI - Successful talc slurry pleurodesis in patients with nonmalignant pleural effusion. AB - BACKGROUND: Chemical pleurodesis is an effective treatment for malignant pleural effusion and pneumothorax. This mode of therapy is, however, less widely accepted in the treatment of patients with refractory benign or undiagnosed pleural effusion. STUDY OBJECTIVES: To analyze the outcome of talc slurry pleurodesis in patients with nonmalignant pleural effusions. DESIGN: Retrospective and partly prospective analysis of clinical outcome. SETTING: Hadassah University Hospital, Jerusalem, Israel. PATIENTS AND PARTICIPANTS: Between 1992 and 1997, we treated 16 patients with nonmalignant pleural effusion using talc slurry pleurodesis. The cause of effusion was congestive heart failure in 6 patients, liver cirrhosis in 4 patients, yellow nail syndrome in 1 patient, systemic lupus erythematosus in 1 patient, chylothorax in 1 patient, and undiagnosed in 3 patients. INTERVENTIONS: Nine patients were hospitalized, and seven patients received treatment in a day care setting. Follow-up ranged from 2 months to 3 years. RESULTS: Complete success was observed in 12 cases (75%), partial success in 3 cases (19%), and pleurodesis was ineffectual in 1 case (6%). There were no significant complications after the procedure in any of our patients. A review of the English language medical literature revealed an additional 110 reported cases of nonmalignant pleural effusion that were treated with chemical pleurodesis. Of these cases, talc was used in 65% with a success rate of nearly 100%. CONCLUSIONS: Chemical pleurodesis, and specifically talc slurry, is an effective treatment for recurrent benign or undiagnosed pleural effusion. This procedure is safe and easily performed and, in selected cases, can be performed in an outpatient day-care setting. PMID- 10807830 TI - Effects of augmented continuous positive airway pressure education and support on compliance and outcome in a Chinese population. AB - OBJECTIVES: To study the effects of augmentation of continuous positive airway pressure (CPAP) education and support on compliance and outcome in patients with obstructive sleep apnea (OSA). DESIGN: A randomized, controlled, parallel study of basic vs augmented CPAP education and support. SETTING: A university teaching hospital. PATIENTS: A total of 108 OSA patients randomized into basic-support (BS) and augmented-support (AS) groups. INTERVENTIONS: Patients in the BS group (n = 54) were given educational brochures on OSA and CPAP, CPAP education by nurses, CPAP acclimatization, and were reviewed by physicians and nurses at weeks 4 and 12. Patients in the AS group (n = 54) received more education, including a videotape, telephone support by nurses, and early review at weeks 1 and 2. MEASUREMENTS: Objective CPAP compliance, Calgary sleep apnea quality of life index (SAQLI), and cognitive function after 1 month and 3 months; and Epworth sleepiness scale (ESS) after 3 months of CPAP treatment. RESULTS: At 4 weeks, CPAP usage was 5.3 +/- 0.2 h/night (mean +/- SEM) vs 5.5 +/- 0.2 h/night in the BS and AS groups, respectively (p = 0.4). At 12 weeks, CPAP usage was 5.3 +/- 0.3 h/night vs 5.3 +/- 0.2 h/night in the two groups, respectively (p = 0.98). There was greater improvement of SAQLI at 4 weeks (p = 0.008) and at 12 weeks (p = 0.047) in the AS group. There was no significant difference between BS and AS groups in terms of improvement of ESS and cognitive function. CONCLUSION: Augmentation of CPAP education and support does not increase CPAP compliance, but leads to a greater improvement of quality of life during the reinforced period. PMID- 10807831 TI - Sleep apnea and hypertension. AB - STUDY OBJECTIVES: To examine the central inspiratory drive response to hypoxia in patients with obstructive sleep apnea (OSA), according to their circadian BP profile, and in healthy control subjects. Another objective was to evaluate the relationships among sleep architecture, hypoxic sensitivity, urinary catecholamine excretion, and BP in OSA patients. PATIENTS AND INTERVENTIONS: Polysomnography, 24-h ambulatory BP recording, and urinary excretion of catecholamines were simultaneously examined in 24 consecutive OSA patients and 11 healthy subjects. OSA patients were categorized as being normotensive (type 1), having BP elevation only during sleep (type 2), and as being hypertensive with elevated BP at all times (type 3). The response of mouth occlusion pressure at 0.1 s after onset (P(0.1)) to progressive isocapnic hypoxic stimulation was measured. RESULTS: There was a significant difference in the P(0.1) response to hypoxia among control subjects ([mean +/- SD] 0.353 +/- 0.129 cm H(2)O/%) and type 1 (0.228 +/- 0.062 cm H(2)O/%), type 2 (0. 345 +/- 0.106 cm H(2)O/%), and type 3 (0.508 +/- 0.118 cm H(2)O/%) OSA patients. In OSA patients, chemosensitivity was related to the apnea-hypopnea index and to the nocturnal excretion of epinephrine. Significant relationships between the nocturnal excretion of epinephrine and BP were noted. On multiple linear regression analysis, the P(0.1) response to hypoxia was the only variable significantly related to diurnal (r(2) = 0.364; p = 0.005) and nocturnal mean BP (r(2) = 0.461; p = 0.002). CONCLUSION: The findings of the present study suggest a possible mediating role of the peripheral chemosensitivity in the association between sleep apnea and hypertension. PMID- 10807832 TI - Safety of bedside percutaneous dilatational tracheostomy in obese patients in the ICU. AB - STUDY OBJECTIVE: To examine the safety of bedside percutaneous dilatational tracheostomy in obese patients. DESIGN: Case series of consecutive obese patients (body mass index > or = 27 kg/m(2)) with acute respiratory failure in a medical, cardiac, or surgical ICU unit who required tracheostomy for failure to wean and continued mechanical ventilatory support. RESULTS: Thirteen obese patients were identified and consented to the procedure. Bedside percutaneous dilatational tracheostomy was successfully performed in the ICU for all 13 patients. Procedural complications were limited to paratracheal tracheostomy tube placement in one patient, with immediate identification and appropriate correction. Postprocedural complications were limited to a cuff leak in one patient. CONCLUSION: Bedside percutaneous tracheostomy can be safely performed in obese patients. PMID- 10807833 TI - Long-term follow-up of Griggs percutaneous tracheostomy with spiral CT and questionnaire. AB - OBJECTIVE: To assess late complications following percutaneous tracheostomy using the Griggs technique. DESIGN: Observational cohort study SETTING: General ICU of a 700-bed district general hospital in the United Kingdom. PATIENTS: Twenty-five patients who underwent Griggs tracheostomy in the ICU and survived for at least 6 months after decannulation. INTERVENTION: Patients were invited to attend for assessment by questionnaire and for spiral CT of the trachea. RESULTS: Eight patients had moderate tracheal dilatation, two patients were permanently hoarse, nine patients had minor voice changes, no patient had tracheal stenosis, and no patient had a disfiguring scar. CONCLUSION: Following Griggs percutaneous tracheostomy, 8 of 25 patients developed moderate tracheal dilatation, and none developed a tracheal stenosis. The cosmetic result after Griggs tracheostomy is good, but the number of patients with minor voice changes is high. A further investigation of long-term outcome following Griggs tracheostomy is necessary. PMID- 10807834 TI - A comparative analysis of patients with early-onset vs late-onset nosocomial pneumonia in the ICU setting. AB - STUDY OBJECTIVE: To compare the clinical outcomes of critically ill patients developing early-onset nosocomial pneumonia (NP; ie, within 96 h of ICU admission) and late-onset NP (ie, occurring after 96 h of ICU admission). DESIGN: Prospective cohort study. SETTING: A medical ICU and a surgical ICU from a university-affiliated urban teaching hospital. PATIENTS: Between July 1997 and November 1998, 3, 668 patients were prospectively evaluated. INTERVENTION: Prospective patient surveillance and data collection. RESULTS: Four hundred twenty patients (11.5%) developed NP. Early-onset NP was observed in 235 patients (56.0%), whereas 185 patients (44.0%) developed late-onset NP. Among patients with early onset NP, 114 patients (48. 5%) spent at least 24 h in the hospital prior to ICU admission, compared to 57 patients (30.8%) with late-onset NP (p = 0.001). One hundred eighty-three patients (77.9%) with early-onset NP received antibiotics prior to the development of NP, as compared to 162 patients (87.6%) with late-onset NP (p = 0.010). The most common pathogens associated with early onset NP were Pseudomonas aeruginosa (25.1%), oxacillin-sensitive Staphylococcus aureus (OSSA; 17.9%), oxacillin-resistant S aureus (ORSA; 17.9%), and Enterobacter species (10.2%). P aeruginosa (38.4%), ORSA (21.1%), Stenotrophomonas maltophilia (11.4%), OSSA (10.8%), and Enterobacter species (10.3%) were the most common pathogens associated with late-onset NP. The ICU length of stay was significantly longer for patients with early-onset NP (10.3 +/ 8.3 days; p < 0.001) and late-onset NP (21. 0 +/- 13.7 days; p < 0.001), as compared to patients without NP (3.5 +/- 3.2 days). Hospital mortality was significantly greater for patients with early-onset NP (37.9%; p = 0.001) and late-onset NP (41.1%; p = 0.001) compared to patients without NP (13.1%). CONCLUSIONS: Both early-onset and late-onset NP are associated with increased hospital mortality rates and prolonged lengths of stay. The pathogens associated with NP were similar for both groups. This may be due, in part, to the prior hospitalization and use of antibiotics in many patients developing early-onset NP. These data suggest that P aeruginosa and ORSA can be important pathogens associated with early-onset NP in the ICU setting. Additionally, clinicians should be aware of the common microorganisms associated with both early-onset NP and late-onset NP in their hospitals in order to avoid the administration of inadequate antimicrobial treatment. PMID- 10807835 TI - Suture or prosthetic reconstruction of experimental diaphragmatic defects. AB - OBJECTIVE: Diaphragmatic reconstruction may cause several respiratory changes. The aims of the present study were to evaluate the respiratory changes induced by two methods of diaphragmatic reconstruction. METHODS: Two groups of rats with an experimental diaphragmatic defect were studied. In one group (n = 5), diaphragmatic resection was followed by stitching together the borders of the wound (SUT); in another group (n = 5), the defect was repaired by suturing in a polytetrafluoroethylene (PTFE) patch. All animals were sedated, anesthetized, paralyzed, and mechanically ventilated. Spirometry, respiratory mechanics, and thoracoabdominal morphometry were evaluated before and after diaphragmatic reconstruction. RESULTS: The suture of the diaphragm significantly decreased FVC and FEV(1), and increased respiratory system, lung, and chest wall static and dynamic elastances and viscoelastic/inhomogeneous pressures in relation to their respective control values. On the other hand, diaphragmatic reconstruction with PTFE increased only respiratory system, lung, and chest wall static elastances. In addition, respiratory system, pulmonary, and chest wall viscoelastic/inhomogeneous pressures and dynamic elastances, as well as respiratory system and lung elastances, were significantly greater in SUT than in PTFE. Lateral diameter at the level of the xiphoid and cephalocaudal pulmonary diameter diminished only in the SUT group. CONCLUSIONS: The reconstruction of the diaphragm with PTFE might be preferred to simple suture for surgical repair of large diaphragmatic defects, at least from a mechanical standpoint. PMID- 10807836 TI - Nd-YAG laser ignition of silicone endobronchial stents. AB - STUDY OBJECTIVES: To test the incendiary characteristics of various silicone endobronchial stents under the impact of the Nd-YAG laser. DESIGN: In vitro study in the laser laboratory of a university-affiliated city hospital. SETTING: The experiments were performed in a reaction chamber under controlled oxygen concentrations. The radiolucent and radiopaque Dumon silicone stent (Novatech; Aubagne, France) and the tracheal part of the Dynamic stent (Ruesch AG; Kernen, Germany) were tested. The Dumon stents were either clean, covered with a thin layer of blood, or mounted on fresh pig tracheal wall. The laser was aimed on them perpendicularly from distances of 1.0 cm and 0.5 cm. INTERVENTIONS: Minimal oxygen concentration to allow ignition and impact time for power outputs (POs) between 10 W and 80 W were determined. MEASUREMENTS AND RESULTS: The lowest oxygen concentration that allowed ignition of some stents was 40%. The clean radiolucent stent could not be ignited with up to 100% ambient oxygen concentration. Radiopacity, presence of blood, tracheal wall, and metal, as well as higher PO and shorter distance of the laser probe decreased impact time to ignition. The radiopaque blood-covered stent was most easily ignited. For this stent, at a PO of 40 W, impact time to ignition was 1.5 +/- 0.2 s, and at 30 W was 2.6 +/- 0.3 s. CONCLUSIONS: At ambient oxygen concentrations > or = 40%, silicone stents can catch fire. Depending on the condition of the stent, the distance of the laser probe, and PO, ignition can occur after short impact times. To prevent stent ignition, oxygen concentration should be kept < 40%. When unusual circumstances require working with higher oxygen concentrations, pulse duration needs to be limited or stent removal might be considered before firing the laser. PMID- 10807837 TI - Phthisiology at the dawn of the new century. AB - Tuberculosis (TB) has been and continues to be one of the most significant pathogens in terms of human morbidity and mortality. Although the resurgence of TB has been held in check in most developed countries, the epidemic rages on in most developing countries of the world. The specter of drug resistance is becoming a more credible challenge in many parts of the world, dimming the prospects of eventual elimination. However, great opportunities are arising as well, with an unprecedented focus on the global aspects of TB control. This article will review the status of TB today and put into perspective the prospects for its elimination in the coming century. PMID- 10807838 TI - End-of-life care preferences of patients enrolled in cardiovascular rehabilitation programs. AB - STUDY OBJECTIVES: The study assessed the interests of ambulatory cardiac patients in advance planning and their willingness to participate in rehabilitation program-based end-of-life education. DESIGN: Observational survey study. SETTING: Fourteen outpatient cardiac rehabilitation programs in 11 states. PARTICIPANTS: Four hundred fifteen subjects enrolled in cardiac rehabilitation. MEASUREMENTS AND RESULTS: A questionnaire determined patient preferences for advance planning, completion of advance directives, completion of patient-physician discussions on end-of-life care, and effects of health status on patient acceptance of life sustaining interventions. Seventy-two percent of patients wanted to direct their own end-of-life care, 86% desired more information on advance directives, 62% wanted to learn about life-sustaining care, and 96% were receptive to advance planning discussions with their physicians. Seventy-two percent of patients had considered that they might require life-sustaining care in the future; acceptability of resuscitative care depended on health status and probability of survival. However, only 15% had discussed advance planning with their physicians, and 10% were confident that their physicians understood their end-of-life wishes. Physicians and cardiovascular rehabilitation programs were considered desirable sources of information on advance planning. CONCLUSIONS: Cardiac patients enrolled in rehabilitation programs want to learn more about end-of-life care and need more opportunities to discuss advance planning with their physicians. Patients consider cardiovascular rehabilitation programs to be acceptable sites for advance planning education. PMID- 10807839 TI - The role of nuclear factor-kappa B in pulmonary diseases. AB - Nuclear factor-kappa B (NF-kappaB) is a family of DNA-binding protein factors that are required for transcription of most proinflammatory molecules, including adhesion molecules, enzymes, cytokines, and chemokines. NF-kappaB activation seems to be a key early event in a variety of cell and animal model systems developed to elucidate the pathobiology of lung diseases. The purpose of this short review is to describe what is known about the molecular biology of NF kappaB and to review information that implicates NF-kappaB in the pathogenesis of lung disease, including ARDS, systemic inflammatory response syndrome, asthma, respiratory viral infections, occupational and environmental lung disease, and cystic fibrosis. PMID- 10807840 TI - Longitudinal changes in bronchial responsiveness associated with swine confinement dust exposure. AB - STUDY OBJECTIVES: Acute exposure to the air in swine confinement units causes bronchial hyperresponsiveness and inflammation of the airways. This study was performed to assess the longitudinal development of bronchial responsiveness in pig farmers and to establish exposure-response relationships. METHODS: A cohort of 171 pig farmers was followed over a 3-year period. Bronchial responsiveness was assessed by a histamine provocation test. Long-term average exposure to inhalable dust and endotoxin was determined by personal monitoring in summer and winter, using data on farm characteristics and activities. Time-weighted average (TWA) personal exposure to ammonia was measured. Data on farm characteristics were gathered in the same period. RESULTS: Mean increase in responsiveness was 2.52 doubling concentrations of histamine for a 10% decrease in FEV(1) and 1.36 doubling concentrations for a 20% decrease in FEV(1). Long-term average exposure to dust was 2.63 mg/m(3) and to endotoxin was 105 ng/m(3). TWA exposure to ammonia was 1.60 mg/m(3). After adjusting for age and smoking behavior, long-term average exposure to inhalable dust was associated with increases in bronchial responsiveness expressed as steps for provocative concentration causing 10% fall in FEV(1). TWA exposure to ammonia, use of wood shavings as bedding, and automated dry feeding were associated with increases in responsiveness expressed as steps for provocative concentration causing a 20% fall in FEV(1). CONCLUSIONS: Exposure to dust and ammonia in pig farms contributes to chronic inflammation of the airways and should be reduced. PMID- 10807841 TI - Rational empiric antibiotic prescription in the ICU. AB - The prescribing of antibiotics in the ICU is usually empiric, given the critical nature of the conditions of patients hospitalized there. Appropriate antibiotic utilization in this setting is crucial not only in ensuring an optimal outcome, but in curtailing the emergence of resistance and containing costs. We propose that research in the ICUs is vitally important in guiding antibiotic prescription practices and, therefore, the achievement of above-stated goals. There is wide institutional diversity in the relative prevalence of predominant pathogens and their antimicrobial susceptibilities, and within individual ICUs there exist unique patient populations with varying risks for and susceptibilities to infections and specific pathogens. Appropriate antibiotic prescription practices should be formulated based on surveillance studies and research at individual ICUs; these goals can be accomplished utilizing existing resources. PMID- 10807842 TI - Two faces of progressive dyspnea. PMID- 10807843 TI - A 63-year-old woman with a 2-month history of dyspnea. PMID- 10807844 TI - Aortic rupture and concomitant transection of the left bronchus after blunt chest trauma. AB - We report a patient with traumatic aortic rupture and preoperatively unrecognized complete disruption of the bronchus for the left lower lobe. Preoperative state was complicated by inadequate oxygenation due to total atelectasis of the unventilated collapsed left lower lobe with consequent significant shunting of the unoxygenated blood. The patient had no massive pneumothorax because the intact peribronchial tissue and pleura covered the injured place, preventing important air leakage. The suspicion of possible concomitant tracheobronchial injury and early diagnostic bronchoscopy are important in patients with aortic rupture after blunt chest trauma. PMID- 10807845 TI - Cardiac tamponade following acupuncture. AB - We present a rare complication of acupuncture in a 83-year-old woman who developed syncope and cardiogenic shock shortly after an acupuncture procedure into the sternum. Echocardiography revealed cardiac tamponade, and pericardiocentesis disclosed hemopericardium. Due to hemodynamic instability, thoracotomy was indicated. A small but actively bleeding perforation of the right ventricle was found and successfully closed. Although acupuncture represents a relatively safe therapeutic intervention, this case report should remind all acupuncturists of possible and sometimes life-threatening adverse effects. PMID- 10807846 TI - Necrotizing pulmonary granulomata in a marijuana smoker. AB - We describe the case of a heavy marijuana and tobacco smoker who presented with progressive exertional dyspnea of 2 months' duration, and bilateral nodular lung infiltrates. Examination of the lung fields was normal, and lung function tests showed mild airflow obstruction with moderately reduced gas transfer. BAL returned green-black fluid consisting predominantly of macrophages laden with carbon pigment. Thoracoscopic lung biopsy showed miliary necrotizing granulomata with an alveolar exudate of carbon-laden macrophages within macroscopically blackened lung. The differential diagnosis of pulmonary granulomata in this patient is discussed. PMID- 10807847 TI - Pneumococcal bacteremia associated with an infected central venous catheter. AB - Pneumococcus (Streptococcus pneumoniae) bacteremia is a serious infection. Pneumococcus has never been implicated as a cause of a central venous catheter related bacteremia. It has been isolated from the catheter tip only twice before, and in one case caused the infection of an infusion port device. We report case of a 41-year-old woman who developed pneumococcal bacteremia after 6 days of an indwelling central venous catheter. The catheter tip grew > 300 cfu of S pneumoniae by the roll-plate method described by Maki and colleagues. No other focus of infection could be found in this patient. To the best of our knowledge, this is the first reported case of pneumococcal bacteremia associated with an infected central venous catheter. PMID- 10807848 TI - Yellow nail syndrome: resolution of yellow nails after successful treatment of breast cancer. AB - Yellow nail syndrome (YNS) is a rare entity of unknown cause in which congenitally hypoplastic lymphatics play a major role in the clinical manifestations of the disease. YNS has been associated with many malignancies and immune disorders. We report a case of new-onset YNS associated with breast cancer and dramatic improvement in the yellow nails with cancer treatment. PMID- 10807849 TI - Myxoid liposarcoma of the supraclavicular fossa. AB - Liposarcomas generally originate most often in the extremities or retroperitoneum, less frequently in the head and neck, and rarely in the thorax. We describe a particularly rare presentation of myxoid liposarcoma originating in the supraclavicular fossa. The mass was resected and has not recurred. We searched our pathology database for other soft-tissue tumors of the supraclavicular fossa and found no other case of sarcoma originating in this site. In addition, we performed a literature review of thoracic and neck liposarcomas to identify similar cases and discuss their clinical course. PMID- 10807850 TI - Successful pulmonary thromboendarterectomy in a patient with Klippel-Trenaunay syndrome. AB - Klippel-Trenaunay syndrome (KTS) is a congenital disorder characterized by a triad of cutaneous vascular nevi, soft tissue or bony hypertrophy, and varicose veins or venous malformations involving one or more extremities. An incidence of venous thromboembolism of up to 22% has been reported in this disorder. Also reported is the development of chronic thromboembolic pulmonary hypertension (CTEPH) and subsequent death from right ventricular failure. We report the first patient with KTS to undergo a successful pulmonary thromboendarterectomy for CTEPH. PMID- 10807851 TI - alpha(1)-adrenoreceptors and pulmonary hypertension. PMID- 10807852 TI - Ehrlichiosis in the United States. PMID- 10807853 TI - What to do when bloody fluid is obtained on pericardiocentesis? PMID- 10807854 TI - Bronchial arterial infusion for lung cancer. PMID- 10807855 TI - Corticosteroids in the emergency treatment of acute severe asthma. PMID- 10807856 TI - Absorption of high-dose inhaled corticosteroids. PMID- 10807857 TI - ECG changes in tension pneumothorax: a hypothesis. PMID- 10807858 TI - Canaries in the coal mine: mitochondrial DNA and vascular injury from reactive oxygen species. PMID- 10807859 TI - Congenital heart defects: trapping the genetic culprits. PMID- 10807860 TI - Leading the way using microarray: a more comprehensive approach for discovery of gene expression patterns. PMID- 10807861 TI - Role of p38 mitogen-activated protein kinases in preconditioning: a detrimental factor or a protective kinase? PMID- 10807862 TI - Human gene therapy: the good, the bad, and the ugly. PMID- 10807863 TI - Ischemic preconditioning in isolated cells. PMID- 10807865 TI - Altered patterns of gene expression in response to myocardial infarction. AB - The use of cDNA microarrays has made it possible to simultaneously analyze gene expression for thousands of genes. Microarray technology was used to evaluate the expression of >4000 genes in a rat model of myocardial infarction. More than 200 genes were identified that showed differential expression in response to myocardial infarction. Gene expression changes were monitored from 2 to 16 weeks after infarction in 2 regions of the heart, the left ventricle free wall and interventricular septum. A novel clustering program was used to identify patterns of expression within this large set of data. Unique patterns were revealed within the transcriptional responses that illuminate changes in biological processes associated with myocardial infarction. PMID- 10807864 TI - Jumonji, a nuclear protein that is necessary for normal heart development. AB - Jumonji (jmj) was cloned in a gene trap screen to identify and mutagenize genes important for heart development. To investigate the role of jmj in heart development, we generated mice homozygous for the jmj mutation. The jmj homozygous mouse embryos showed heart malformations, including ventricular septal defect, noncompaction of the ventricular wall, double-outlet right ventricle, and dilated atria. The jmj mutants died soon after birth, apparently as a result of respiratory insufficiency caused by rib and sternum defects in addition to the heart defects. In situ hybridization analyses suggested that cardiomyocytes were differentiated but developmental regulation of chamber-specific genes was defective in fetal hearts. Expression of jmj was detected in the myocardium, especially in the interventricular septum, ventricular wall, and outflow tract, which correlated well with the locations of defects observed in the hearts of mutant mice. Homozygous embryos failed to express the jmj transcript in all tissues except in the nervous system. Confocal microscopic examination using anti JMJ antibodies indicated that the JMJ protein was localized in the nuclei of cells transfected with jmj. These data demonstrate that JMJ is a nuclear protein, which is essential for normal heart development and function. PMID- 10807866 TI - Expression, release, and biological activity of parathyroid hormone-related peptide from coronary endothelial cells. AB - Ventricular cardiomyocytes have previously been identified as potential target cells for parathyroid hormone-related peptide (PTHrP). Synthetic PTHrP peptides exert a positive contractile effect. Because systemic PTHrP levels are normally negligible, this suggests that PTHrP is expressed in the ventricle and acts as a paracrine mediator. We investigated the ventricular expression of PTHrP and its expression in cultured cells isolated from the ventricle, studied the release of PTHrP from hearts and cultures, and investigated whether this authentic PTHrP mimics the biological effects previously described for synthetic PTHrP on ventricular cardiomyocytes. We found PTHrP expressed in ventricles of neonatal and adult rat hearts. In cells isolated from adult hearts, we found PTHrP expression exclusively in coronary endothelial cells but not in cardiomyocytes. The latter, however, are target cells for PTHrP. PTHrP was released from isolated perfused hearts during hypoxic perfusion and from cultured coronary endothelial cells under energy-depleting conditions. This PTHrP was biologically active; ie, it exerted a positive contractile and lusitropic effect on cardiomyocytes. Authentic PTHrP was glycosylated and showed a slightly higher potency than synthetic PTHrP. These results suggest that PTHrP is an endothelium-derived modulator of ventricular function. PMID- 10807867 TI - Angiopoietin-1 induces endothelial cell sprouting through the activation of focal adhesion kinase and plasmin secretion. AB - Angiopoietin-1 (Ang1) is a strong inducer of endothelial cell sprouting, which is a first step in both angiogenesis and neovascularization. We examined the mechanisms underlying Ang1-induced cell sprouting using porcine pulmonary artery endothelial cells. Ang1 induced the nondirectional and directional migration of endothelial cells mediated through the Tie2 but not the Tie1 receptor. Ang1 induced tyrosine phosphorylation of p125(FAK), and this phosphorylation was dependent on phosphatidylinositol (PI) 3'-kinase activity. Ang1 induced the secretion of plasmin and matrix metalloproteinase-2 (MMP-2), which is inhibited by PI 3'-kinase inhibitors. Ang1 also induced the secretion of small amounts of proMMP-3 and proMMP-9 but not proMMP-1. Ang1 suppressed the secretion of tissue inhibitor of metalloproteinase-2 (TIMP-2), but not of TIMP-1. Addition of alpha(2)-antiplasmin, a combination of TIMP-1 and TIMP-2, or PI 3'-kinase inhibitors inhibited Ang1-induced sprouting activity. Therefore, Ang1-induced sprouting activity in endothelial cells may be accomplished by cytoskeletal changes and secretion of proteinases and may be largely mediated through intracellular PI 3'-kinase activation. PMID- 10807868 TI - Hydrogen peroxide- and peroxynitrite-induced mitochondrial DNA damage and dysfunction in vascular endothelial and smooth muscle cells. AB - The mechanisms by which reactive species (RS) participate in the development of atherosclerosis remain incompletely understood. The present study was designed to test the hypothesis that RS produced in the vascular environment cause mitochondrial damage and dysfunction in vitro and, thus, may contribute to the initiating events of atherogenesis. DNA damage was assessed in vascular cells exposed to superoxide, hydrogen peroxide, nitric oxide, and peroxynitrite. In both vascular endothelial and smooth muscle cells, the mitochondrial DNA (mtDNA) was preferentially damaged relative to the transcriptionally inactive nuclear beta-globin gene. Similarly, a dose-dependent decrease in mtDNA-encoded mRNA transcripts was associated with RS treatment. Mitochondrial protein synthesis was also inhibited in a dose-dependent manner by ONOO(-), resulting in decreased cellular ATP levels and mitochondrial redox function. Overall, endothelial cells were more sensitive to RS-mediated damage than were smooth muscle cells. Together, these data link RS-mediated mtDNA damage, altered gene expression, and mitochondrial dysfunction in cell culture and reveal how RS may mediate vascular cell dysfunction in the setting of atherogenesis. PMID- 10807869 TI - Fluid shear stress induces lipocalin-type prostaglandin D(2) synthase expression in vascular endothelial cells. AB - Ligands for peroxisome proliferator-activated receptor gamma, such as the thiazolidinedione class of antidiabetic drugs and 15-deoxy-Delta(12,14) prostaglandin J(2) (15d-PGJ(2)), modulate various processes in atherogenesis. In search of cells that generate prostaglandin D(2) (PGD(2)), the metabolic precursor of 15d-PGJ(2), we identified PGD(2) from culture medium of endothelial cells. To study how PGD(2) production is regulated in endothelial cells, we investigated the role of fluid shear stress in the metabolism of PGD(2). Endothelial cells expressed the mRNA for the lipocalin-type PGD(2) synthase (L PGDS) both in vitro and in vivo. Loading laminar shear stress using a parallel plate flow chamber markedly enhanced the gene expression of L-PGDS, with the maximal effect being obtained at 15 to 30 dyne/cm(2). The expression began to increase within 6 hours after loading shear stress and reached the maximal level at 18 to 24 hours. In contrast, shear stress did not alter the expression levels of PGI(2) synthase and thromboxane A(2) synthase. In parallel with the increase in the expression level of L-PGDS, endothelial cells released PGD(2) and 15d PGJ(2) into culture medium. These results demonstrate that shear stress promotes PGD(2) production by stimulating L-PGDS expression and suggest the possibility that a peroxisome proliferator-activated receptor gamma ligand is produced in vascular wall in response to blood flow. PMID- 10807870 TI - Hypoxia-induced endothelial apoptosis through nuclear factor-kappaB (NF-kappaB) mediated bcl-2 suppression: in vivo evidence of the importance of NF-kappaB in endothelial cell regulation. AB - The transcription factor nuclear factor-kappaB (NF-kappaB) plays a pivotal role in the coordinated transactivation of cytokine and adhesion molecule genes involved in endothelial activation. Although recent reports have documented the contribution of NF-kappaB to apoptosis, it is still controversial. Especially, the role of NF-kappaB in endothelial apoptosis is largely unknown. Hypoxia significantly induced human aortic endothelial cell death and apoptosis in a time dependent manner (P<0.01), accompanied by NF-kappaB activation. Decrease in total cell number and increase in apoptotic cells induced by hypoxia were significantly attenuated by NF-kappaB decoy, but not by scrambled decoy, oligodeoxynucleotides (ODNs) (P<0.01). Increase in DNA fragmentation induced by hypoxia was also significantly inhibited by NF-kappaB decoy ODNs as compared with scrambled decoy ODNs (P<0.01). Moreover, transfection of NF-kappaB decoy ODNs resulted in a significant decrease in caspase-3-like activity, which is a common pathway for apoptosis, compared with scrambled decoy ODNs. Importantly, transfection of NF kappaB decoy ODNs significantly increased protein of bcl-2, an inhibitor of apoptosis, and did not alter bax, a promoter of apoptosis, thereby resulting in a significant increase in the ratio of bcl-2 to bax (P<0.01). bcl-2 mRNA was also decreased by hypoxia, whereas transfection of NF-kappaB decoy ODNs significantly attenuated decrease in bcl-2 mRNA. These results demonstrate that activation of NF-kappaB by hypoxia induced endothelial apoptosis in a bcl-2-dependent manner. The importance of NF-kappaB in endothelial apoptosis was confirmed by the observation that pyrrolidine dithiocarbamate, a potent NF-kappaB inhibitor, prevented endothelial apoptosis, caspase 3-like activity, and bcl-2 downregulation induced by hypoxia. To test this hypothesis in vivo, we transfected NF-kappaB decoy ODNs into rat intact carotid artery after reperfusion injury. Reperfusion injury was associated with a significant increase in endothelial apoptosis at 24 hours, whereas NF-kappaB decoy ODN treatment markedly decreased terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL)-positive endothelial cells at 24 hours after reperfusion (P<0.01). Here, using synthetic double-stranded DNA with high affinity for NF-kappaB as a decoy approach, we demonstrated that activation of NF-kappaB by hypoxia caused aortic endothelial cell death and apoptosis through the suppression of bcl-2. NF-kappaB mediated endothelial apoptosis induced by hypoxia may be involved in the pathogenesis of endothelial dysfunction observed in cardiovascular ischemic diseases. PMID- 10807871 TI - cAMP pulse during preservation inhibits the late development of cardiac isograft and allograft vasculopathy. AB - The causes of transplant-associated coronary artery disease remain obscure, and there is no known treatment. Preservation injury of murine heterotopic vascularized cardiac isografts caused a small, albeit significant, increase in neointimal formation; preservation injury of allografts markedly increased both the incidence and severity of transplant-associated coronary artery disease. As cAMP is an important vascular homeostatic mediator the levels of which decline during organ preservation, buttressing cAMP levels solely during initial preservation both improved acute allograft function and reduced the severity of transplant-associated coronary artery disease in grafts examined 2 months later. Inhibiting the cAMP-dependent protein kinase abrogated these beneficial effects. cAMP treatment was associated with an early reduction in leukocyte infiltration and a reciprocal decrease in superoxide and increase in NO levels. These data indicate that alloantigen-independent injury to the graft, which occurs at the time of cardiac preservation, can set in motion pathological vascular events that are manifest months later. Furthermore, a cAMP pulse during cardiac preservation reduces the incidence and severity of transplant-associated coronary artery disease. PMID- 10807872 TI - Adenosine A(1) receptor induced delayed preconditioning in rabbits: induction of p38 mitogen-activated protein kinase activation and Hsp27 phosphorylation via a tyrosine kinase- and protein kinase C-dependent mechanism. AB - Transient adenosine A(1) receptor (A(1)R) activation in rabbits induces delayed preconditioning against myocardial infarction 24 to 72 hours later. The cellular mechanisms downstream of A(1)R mediating this delayed cardioprotection have not been elucidated. This study examined the role of protein kinase C (PKC) and tyrosine kinases (TKs) in the signaling cascade mediating A(1)R-induced late preconditioning in rabbits. The small heat shock protein Hsp27 has been shown to confer cytoskeletal protection when in the phosphorylated state. We therefore also evaluated the potential role of the p38 mitogen-activated protein kinase (p38 MAPK) and Hsp27 as distal mediators of A(1)R-induced delayed preconditioning. Pharmacological preconditioning of rabbits with the selective A(1) agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA; 100 microgram/kg) significantly reduced myocardial infarct size compared with control animals, after 30-minute regional ischemia/2-hour reperfusion in vivo 24 hours later (23.7+/-3.1 versus 43.0+/-4.1%; P<0.05). This delayed protection was abrogated by prior inhibition of either PKC with chelerythrine chloride (5 mg/kg) or of TKs with lavendustin A (1.3 mg/kg), suggesting that both PKC and TK are crucial for the development of delayed preconditioning after A(1) receptor activation in the rabbit. Myocardial tissue extracts obtained 24 hours after CCPA treatment were analyzed for p38 MAPK catalytic activity using an in vitro kinase assay. This showed an almost 7-fold increase in p38 MAPK activity in myocardial samples pretreated with CCPA compared with control hearts. Two-dimensional gel electrophoresis revealed an increase in the phosphorylated isoforms of Hsp27 in hearts pretreated with CCPA compared with control hearts. Prior inhibition of either PKC or TK prevented the CCPA-induced increase in p38 MAPK activity and phosphorylation of Hsp27. This study identifies new components of the signaling mechanism of A(1)R-induced delayed preconditioning. Our results suggest an important role for both PKC and TK as mediators of late preconditioning against infarction after A(1)R activation and, although correlative, point to the p38 MAPK/Hsp27 pathway as a potential distal effector of this protection. PMID- 10807873 TI - Allele-specific regulation of matrix metalloproteinase-12 gene activity is associated with coronary artery luminal dimensions in diabetic patients with manifest coronary artery disease. AB - Both the processes of atherosclerosis and plaque rupture are indicated to be influenced by matrix metalloproteinase (MMP) activity. We therefore searched for common functional variation in the matrix metalloelastase (MMP-12) gene locus that may be implicated in coronary artery disease. Single-strand conformation polymorphism analysis of DNA from healthy individuals detected a common polymorphism within the MMP-12 gene promoter (an A-to-G substitution at position 82). The frequency of the G allele was 0. 19. The polymorphism influences the binding of the transcription factor activator protein-1 (AP-1) in electromobility shift assay. A higher binding affinity of AP-1 to the A allele was associated with higher MMP-12 promoter activity in vitro in transient transfection studies in U937 and murine lung macrophage (MALU) cells. Phorbol 12-myristate 13-acetate (PMA) and insulin, 2 known activators of AP-1, increased the binding of AP-1 to the MMP-12 promoter, with higher affinity for the A allele. In transfection experiments, both the A and the G alleles responded to insulin and PMA, the A allele showing higher promoter activity than the G allele. Furthermore, Western blot analysis demonstrated that insulin increased MMP-12 protein production. To analyze whether the -82 A/G polymorphism is associated with coronary artery disease, 367 consecutive patients who underwent percutaneous transluminal coronary angiography with stent implantation were genotyped. In patients (n=71) with diabetes, the A allele was associated with a smaller luminal diameter. In conclusion, a common functional polymorphism within the MMP-12 promoter influences coronary artery luminal dimensions in diabetic patients with manifest coronary artery disease. PMID- 10807874 TI - Activation of mitogen-activated protein kinases in human heart during cardiopulmonary bypass. AB - Mitogen-activated protein kinases (MAPKs) have been shown to be activated in both in vitro and in vivo models of cardiac tissue in response to ischemia/reperfusion injury. We investigated whether MAPKs are activated in human heart during coronary artery bypass grafting (CABG) surgery. During elective CABG surgery of 8 patients, 3 right atrial appendage biopsies were obtained at baseline, at the end of cross-clamping, and after coronary reperfusion. The expression of the p38 MAPK, c-Jun N-terminal kinase (JNK), and extracellular signal-regulated kinases (ERK1/2) MAPKs was not altered during CABG. The phosphorylation and activation of both ERK1/2 and p38-MAPK were increased approximately 2-fold by ischemia and even more (8- and 4-fold, respectively) by reperfusion. Although the ischemic period did not result in a significant activation of JNK, an approximately 6-fold increase in JNK activity could be observed after reperfusion. In conclusion, distinct activation patterns of ERK1/2, p38, and JNK MAPKs can be observed in human heart during CABG. PMID- 10807875 TI - UltraRapid communications : vascular superoxide production by NAD(P)H OxidaseAssociation with endothelial dysfunction and clinical risk factors AB - Superoxide anion plays important roles in vascular disease states. Increased superoxide production contributes to reduced nitric oxide (NO) bioactivity and endothelial dysfunction in experimental models of vascular disease. We measured superoxide production by NAD(P)H oxidase in human blood vessels and examined the relationships between NAD(P)H oxidase activity, NO-mediated endothelial function, and clinical risk factors for atherosclerosis. Endothelium-dependent vasorelaxations and direct measurements of vascular superoxide production were determined in human saphenous veins obtained from 133 patients with coronary artery disease and identified risk factors. The predominant source of vascular superoxide production was an NAD(P)H-dependent oxidase. Increased vascular NAD(P)H oxidase activity was associated with reduced NO-mediated vasorelaxation. Furthermore, reduced endothelial vasorelaxations and increased vascular NAD(P)H oxidase activity were both associated with increased clinical risk factors for atherosclerosis. Diabetes and hypercholesterolemia were independently associated with increased NADH-dependent superoxide production. The association of increased vascular NAD(P)H oxidase activity with endothelial dysfunction and with clinical risk factors suggests an important role for NAD(P)H oxidase-mediated superoxide production in human atherosclerosis. The full text of this article is available at http://www.circresaha.org. Key Words:atherosclerosis endothelium superoxide nitric oxide diabetes Two Distinct Congenital Arrhythmias Evoked by a Multidysfunctional Na(+) Channel Marieke W. Veldkamp, Prakash C. Viswanathan, Connie Bezzina, Antonius Baartscheer, Arthur A.M. Wilde, Jeffrey R. Balser Abstract-The congenital long-QT syndrome (LQT3) and the Brugada syndrome are distinct, life-threatening rhythm disorders linked to autosomal dominant mutations in SCN5A, the gene encoding the human cardiac Na(+) channel. It is believed that these two syndromes result from opposite molecular effects: LQT3 mutations induce a gain of function, whereas Brugada syndrome mutations reduce Na(+) channel function. Paradoxically, an inherited C-terminal SCN5A mutation causes affected individuals to manifest electrocardiographic features of both syndromes: QT-interval prolongation (LQT3) at slow heart rates and distinctive ST segment elevations (Brugada syndrome) with exercise. In the present study, we show that the insertion of the amino acid 1795insD has opposite effects on two distinct kinetic components of Na(+) channel gating (fast and slow inactivation) that render unique, simultaneous effects on cardiac excitability. The mutation disrupts fast inactivation, causing sustained Na(+) current throughout the action potential plateau and prolonging cardiac repolarization at slow heart rates. At the same time, 1795insD augments slow inactivation, delaying recovery of Na(+) channel availability between stimuli and reducing the Na(+) current at rapid heart rates. Our findings reveal a novel molecular mechanism for the Brugada syndrome and identify a new dual mechanism whereby single SCN5A mutations may evoke multiple cardiac arrhythmia syndromes by influencing diverse components of Na(+) channel gating function. The full text of this article is available at http://www.circresaha.org. Key Words: Na(+) channel inactivation long-QT syndrome Brugada syndrome PMID- 10807876 TI - Vascular superoxide production by NAD(P)H oxidase: association with endothelial dysfunction and clinical risk factors. AB - Superoxide anion plays important roles in vascular disease states. Increased superoxide production contributes to reduced nitric oxide (NO) bioactivity and endothelial dysfunction in experimental models of vascular disease. We measured superoxide production by NAD(P)H oxidase in human blood vessels and examined the relationships between NAD(P)H oxidase activity, NO-mediated endothelial function, and clinical risk factors for atherosclerosis. Endothelium-dependent vasorelaxations and direct measurements of vascular superoxide production were determined in human saphenous veins obtained from 133 patients with coronary artery disease and identified risk factors. The predominant source of vascular superoxide production was an NAD(P)H-dependent oxidase. Increased vascular NAD(P)H oxidase activity was associated with reduced NO-mediated vasorelaxation. Furthermore, reduced endothelial vasorelaxations and increased vascular NAD(P)H oxidase activity were both associated with increased clinical risk factors for atherosclerosis. Diabetes and hypercholesterolemia were independently associated with increased NADH-dependent superoxide production. The association of increased vascular NAD(P)H oxidase activity with endothelial dysfunction and with clinical risk factors suggests an important role for NAD(P)H oxidase-mediated superoxide production in human atherosclerosis. The full text of this article is available at http://www.circresaha.org. PMID- 10807877 TI - Two distinct congenital arrhythmias evoked by a multidysfunctional Na(+) channel. AB - The congenital long-QT syndrome (LQT3) and the Brugada syndrome are distinct, life-threatening rhythm disorders linked to autosomal dominant mutations in SCN5A, the gene encoding the human cardiac Na(+) channel. It is believed that these two syndromes result from opposite molecular effects: LQT3 mutations induce a gain of function, whereas Brugada syndrome mutations reduce Na(+) channel function. Paradoxically, an inherited C-terminal SCN5A mutation causes affected individuals to manifest electrocardiographic features of both syndromes: QT interval prolongation (LQT3) at slow heart rates and distinctive ST-segment elevations (Brugada syndrome) with exercise. In the present study, we show that the insertion of the amino acid 1795insD has opposite effects on two distinct kinetic components of Na(+) channel gating (fast and slow inactivation) that render unique, simultaneous effects on cardiac excitability. The mutation disrupts fast inactivation, causing sustained Na(+) current throughout the action potential plateau and prolonging cardiac repolarization at slow heart rates. At the same time, 1795insD augments slow inactivation, delaying recovery of Na(+) channel availability between stimuli and reducing the Na(+) current at rapid heart rates. Our findings reveal a novel molecular mechanism for the Brugada syndrome and identify a new dual mechanism whereby single SCN5A mutations may evoke multiple cardiac arrhythmia syndromes by influencing diverse components of Na(+) channel gating function. The full text of this article is available at http://www.circresaha.org. PMID- 10807878 TI - Hepatic stem cells. PMID- 10807879 TI - Colorectal cancer screening in the UK: Joint Position Statement by the British Society of Gastroenterology, The Royal College of Physicians, and The Association of Coloproctology of Great Britain and Ireland. PMID- 10807880 TI - Perioperative nutritional support: does it reduce hospital complications or shorten convalescence? PMID- 10807881 TI - Morphine and biliary pain revisited. PMID- 10807882 TI - Innate immunity to carcinomas? PMID- 10807883 TI - Symptomatic gastro-oesophageal reflux as a risk factor for oesophageal adenocarcinoma. PMID- 10807884 TI - Neuromuscular function of the human lower oesophageal sphincter in reflux disease and Barrett's oesophagus. AB - BACKGROUND: Columnar lined (Barrett's) oesophagus is often considered a sequel to chronic severe reflux disease. Aberrant lower oesophageal sphincter (LOS) motility associated with Barrett's oesophagus includes reduced basal LOS pressures. The aim of this study was to characterise neuromuscular function of the LOS in normal (squamous cell carcinoma (SCC) with uninvolved LOS) and reflux affected (Barrett's) oesophagus in vitro. METHODS: Strips of LOS muscle were prepared at biopsy following oesophagectomy from 16 patients with SCC and seven patients with oesophageal adenocarcinoma and Barrett's oesophagus associated with a history of reflux disease. LOS smooth muscle responses were recorded in response to electrical field stimulation (EFS), potassium chloride (KCl), DMPP, isoprenaline, capsaicin, bethanechol, and tachykinins. RESULTS: Basal LOS tone and LOS relaxations in response to isoprenaline, EFS, and DMPP were not significantly altered in the Barrett's group. After tetrodotoxin pretreatment, responses to KCl and DMPP were significantly reduced in the SCC but not in Barrett's LOS. Maximal contraction in response to bethanechol was significantly decreased in Barrett's LOS while substance P and NK-2 receptor mediated contraction was unaltered. Capsaicin, NK-1, and NK-3 receptor agonists exerted negligible effects on LOS tone. CONCLUSIONS: LOS muscle strips from patients with reflux associated Barrett's oesophagus exhibit a reduction in cholinergic muscle contraction while retaining similar features of basal tone, responses to tachykinins, and inhibitory muscle and neural function. Enteric inhibitory neurones in LOS muscle strips from patients with reflux associated Barrett's oesophagus display resistance to axonal sodium channel blockade. No evidence for functional NK-1 or NK-3 receptors or capsaicin sensitive axon collateral reflexes was observed in the human LOS. PMID- 10807885 TI - Cough threshold in reflux oesophagitis: influence of acid and of laryngeal and oesophageal damage. AB - BACKGROUND: Gastro-oesophageal reflux is often associated with cough. Patients with reflux show an enhanced tussive response to bronchial irritants, even in the absence of respiratory symptoms. AIM: To investigate the effect of mucosal damage (either oesophageal or laryngeal) and of oesophageal acid flooding on cough threshold in reflux patients. PATIENTS: We studied 21 patients with reflux oesophagitis and digestive symptoms. Respiratory diseases, smoking, and use of drugs influencing cough were considered exclusion criteria. METHODS: Patients underwent pH monitoring, manometry, digestive endoscopy, laryngoscopy, and methacholine challenge. We evaluated the cough response to inhaled capsaicin (expressed as PD5, the dose producing five coughs) before therapy, after five days of omeprazole therapy, and when oesophageal and laryngeal damage had healed. RESULTS: In all patients spirometry and methacholine challenge were normal. Thirteen patients had posterior laryngitis and eight complained of coughing. Twenty patients showed an enhanced cough response (basal PD5 0.92 (0.47) nM; mean (SEM)) which improved after five and 60 days (2.87 (0.82) and 5.88 (0.85) nM; p<0.0001). The severity of oesophagitis did not influence PD5 variation. On the contrary, the response to treatment was significantly different in patients with and without laryngitis (p = 0.038). In patients with no laryngitis, the cough threshold improved after five days with no further change thereafter. In patients with laryngitis, the cough threshold improved after five days and improved further after 60 days. Proximal and distal oesophageal acid exposure did not influence PD5. Heartburn disappeared during the first five days but the decrease in cough and throat clearing were slower. CONCLUSIONS: Patients with reflux oesophagitis have a decreased cough threshold. This is related to both laryngeal inflammation and acid flooding of the oesophagus but not to the severity of oesophagitis. Omeprazole improves not only respiratory and gastro-oesophageal symptoms but also the cough threshold. PMID- 10807886 TI - A valid, accurate, office based non-radioactive test for gastric emptying of solids. AB - BACKGROUND: Current breath tests for measurement of gastric emptying of solids are expensive, possibly inaccurate, and require cumbersome calculations. AIMS: We wished to validate a simplified solid gastric emptying test using a [(13)C]Spirulina platensis breath test for accurate results relative to scintigraphy. SUBJECTS: Thirty healthy volunteers. METHODS: We measured gastric emptying of egg containing [(13)C]S platensis and (99m)Tc sulphur colloid by breath (13)CO(2) and scintigraphy over six hours. A generalised linear regression model was used to predict t(1/2) and t(LAG) by scintigraphy from breath (13)CO(2) data. The model was cross validated and normative data calculated for a prepacked [(13)C]meal. RESULTS: Regression models using all breath data over six hours, for the first three hours, and for samples at 75, 90, and 180 minutes ("reduced model") predicted t(1/2) and t(LAG) values similar to scintigraphy (t(LAG) 43 (SD 12) min; t(1/2) 100 (20) min). Standard deviations of differences in t(1/2) and t(LAG) between scintigraphy and the "reduced model" were both 10 minutes. Gastric t(1/2) for the prepacked [(13)C]meal was 91 (15) min (10-90% range: 74-118). CONCLUSION: The [(13)C]S platensis breath test and a simple formula using breath (13)CO(2) at baseline, 90, and 180 minutes measured gastric emptying t(1/2) for solids with results that were comparable with scintigraphy. PMID- 10807887 TI - Role of lipase in the regulation of postprandial gastric acid secretion and emptying of fat in humans: a study with orlistat, a highly specific lipase inhibitor. AB - BACKGROUND AND AIMS: To investigate the importance of lipase on gastric functions, we studied the effects of orlistat, a potent and specific inhibitor of lipase, on postprandial gastric acidity and gastric emptying of fat. METHODS: Fourteen healthy volunteers participated in a double blind, placebo controlled, randomised study. In a two way cross over study with two test periods of five days, separated by at least 14 days, orlistat 120 mg three times daily or placebo was given with standardised daily meals. In previous experiments we found that this dose almost completely inhibited postprandial duodenal lipase activity. Subjects underwent 28 hour intragastric pH-metry on day 4, and a gastric emptying study with a mixed meal (800 kcal) labelled with (999m)Tc sulphur colloid (solids) and (111In)thiocyanate (fat) on day 5. Gastric pH data were analysed for three postprandial hours and the interdigestive periods. RESULTS: Orlistat inhibited almost completely (by 75%) lipase activity and accelerated gastric emptying of both the solid (by 52%) and fat (by 44%) phases of the mixed meal (p<0.03). Orlistat increased postprandial gastric acidity (from a median pH of 3.3 to 2.7; p<0.01). Postprandial cholecystokinin release was lower with orlistat (p<0.03). CONCLUSION: Lipase has an important role in the regulation of postprandial gastric acid secretion and fat emptying in humans. These effects might be explained by lipolysis induced release of cholecystokinin. PMID- 10807888 TI - Localisation of cyclooxygenase 1 and cyclooxygenase 2 in Helicobacter pylori related gastritis and gastric ulcer tissues in humans. AB - BACKGROUND: Prostaglandin endoperoxide synthase/cyclooxygenase (COX) is the key enzyme in gastric mucosal protection and repair but its cellular localisation in the human stomach is still unclear. AIMS: To investigate immunohistochemically the cellular distribution of COX-1 and COX-2 proteins in the human stomach with or without gastritis or ulceration. PATIENTS AND METHODS: Tissues were obtained by surgical resection of gastric ulcers associated with perforation (n = 9) or by biopsy from Helicobacter pylori positive patients with gastric ulcers (n = 45) and H pylori negative healthy subjects (n = 15). COX expression was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), western blotting, and light and electron microscopic immunohistochemistry. RESULTS: COX-2 mRNA and protein were detected in gastric ulcer tissues but not in intact gastric mucosa. COX-1 mRNA and protein were detected in the intact mucosa. COX-2 immunostaining was exclusively localised in macrophages and fibroblasts between necrotic and granulation tissues of the ulcer bed. The percentage of COX 2 expressing cells was significantly higher in open than in closed ulcers, and in gastritis than in gastric mucosa without H pylori infection. COX-1 immunoreactivity localised in lamina propria mesenchymal cells was similar in various stages of ulcer disease and in intact gastric mucosa. Electron microscopic immunohistochemistry revealed both COX-1 and COX-2 on the luminal surfaces of the endoplasmic reticulum and nuclear envelope of macrophages and fibroblasts. CONCLUSIONS: Our results showed that COX-2 protein was induced in macrophages and fibroblasts in gastric ulcers and H pylori related gastritis, suggesting its involvement in the tissue repair process. PMID- 10807889 TI - Microsatellite instability in patients with multiple primary cancers of the gastrointestinal tract. AB - BACKGROUND: Little is known about the genetic alterations in multiple primary cancers of the gastrointestinal tract. Microsatellite instability (MSI) is frequently observed in hereditary non-polyposis colorectal cancer (HNPCC), and multiple primary cancers is a feature of this syndrome. AIMS: To identify MSI incidence, target gene mutation, and mismatch repair (MMR) protein status in patients with multiple primary cancers of the gastrointestinal tract. SUBJECTS: Fifty seven cancers from 22 Japanese patients with multiple primary cancers of the stomach, duodenum, colon, and rectum. METHODS: MSI was examined at 5-7 microsatellite loci. Mutation analysis for TGFbetaRII, IGFIIR, and BAX was performed in cancers with MSI. MMR protein status was examined by immunohistochemical analysis using a monoclonal antibody against hMSH2 and hMLH1. RESULTS: MSI was observed in 16 of 22 patients (73%) and in 29 of 57 lesions (51%). High frequency MSI (MSI-H) was found often in patients with multiple cancers in the same organ (p = 0.042), especially in multiple gastric cancer patients (p = 0.038). In contrast, patients with multiple cancers in different organs had a tendency to show low frequency MSI (MSI-L) or microsatellite stable (MSS) phenotype. Both target gene mutation and decreased expression of MMR protein were found only in seven lesions of three patients with MSI at more than four microsatellite loci. CONCLUSIONS: These results suggest that genetic instability may play an important role in the development of multiple gastrointestinal cancers but there may be different genetic alterations between multiple gastrointestinal cancers of the same and different organs. PMID- 10807890 TI - Faecal calprotectin levels in a high risk population for colorectal neoplasia. AB - BACKGROUND: Faecal concentrations of the protein calprotectin have been found to be elevated in patients with colorectal neoplasia, suggesting that it might be used as a screening tool for colorectal cancer as well as adenomas. AIMS: To measure the sensitivity and specificity of faecal calprotectin for the detection of adenomas in high risk individuals undergoing colonoscopy. Also, to investigate between and within stool variability of calprotectin concentrations. SUBJECTS: A total of 814 patients planned for colonoscopy were included for the following indications: positive faecal occult blood test, 25; neoplasia surveillance, 605; newly detected polyp, 130; and family risk, 54. METHODS: Two faecal samples from each of two stools were analysed using the PhiCal ELISA test device (Nycomed Pharma AS). RESULTS: Adenoma patients had significantly higher calprotectin levels than normal subjects (median 9.1 (95% confidence interval 7.5-10.1) v 6.6 (5.6-7.4)mg/l). There was no significant decrease in calprotectin levels after polypectomy. Levels in cancer patients were significantly higher than those in all other subgroups (median 17.6 mg/l (11.5-31.0)). With a cut off limit of 10 mg/l, the sensitivity for cancer was 74% and for adenoma 43%. Corresponding specificity values were 64% for no cancer and 67% for no neoplasia (cancer+adenoma). Specificity varied from 71% for one stool sample to 63% for four samples. Stool variability was small, suggesting that two spots from one stool were as discriminative as two spots from each of two stools. CONCLUSIONS: The sensitivity and specificity of faecal calprotectin levels as a marker for colorectal adenoma and carcinoma justifies its use in high risk groups, but specificity is too low for screening of average risk persons. Lack of a decrease in levels after polypectomy may be due to a more widespread leucocyte migration into the intestinal lumen than that at the polyp site, and needs further investigation. PMID- 10807891 TI - A new variable stiffness colonoscope makes colonoscopy easier: a randomised controlled trial. AB - BACKGROUND: Colonoscopy remains technically difficult in 10-20% of procedures due to variable colonic anatomy and fixation. The ability to vary endoscope shaft flexibility may help insertion to the caecum. METHODS: Consecutive patients attending for day case colonoscopy were randomised to examination with either the conventional Olympus CF200HL (200HL) or a new variable stiffness (VS) colonoscope. Intubation time, use of stiffening function, and patient pain scores were compared. RESULTS: Of 100 cases, 43 were performed with the 200HL and 57 with the VS. Four incomplete examinations occurred with the 200HL (two sigmoid fixations, two benign strictures) and two with the VS (one obstructing cancer, one fixed sigmoid). Changing to the paediatric scope was successful in all but one patient from each group (obstructive lesions). Stiff mode was applied 23 times in 18 patients and was effective in 15 of these. Intubation time was quicker with the VS (median 6 minutes 32 seconds) than with the 200HL (median 10 minutes 35 seconds) (p = 0.0005). Pain scores were less with the VS (median 7) than with the 200HL (median 24) (p = 0.0081). CONCLUSIONS: The variable stiffness colonoscope combines paediatric shaft characteristics with the ability to stiffen when needed. This instrument significantly reduces intubation time and patient discomfort. Further comparisons should be made with the newest colonoscopes which are less stiff. PMID- 10807893 TI - A randomised controlled trial evaluating the use of enteral nutritional supplements postoperatively in malnourished surgical patients. AB - BACKGROUND: Patients who undergo surgery are at risk of malnutrition due to periods of starvation, the stress of surgery, and subsequent increase in metabolic rate. There are limited data on nutritional outcome of surgical patients. AIMS: To investigate changes in nutritional status and the influence of oral supplements on nutritional status, morbidity, and quality of life in postoperative surgical patients. METHODS: Entry was determined by the presence of malnutrition, as defined by a body mass index (BMI) < or =20 kg/m(2), anthropometric measurements < or =15th percentile on admission, or initiation of oral diet postoperatively and/or a weight loss of 5% or more during the operative period. We studied 101 patients: 52 were randomised to the treatment group (TG) and prescribed a 1.5 kcal/ml nutritional supplement; 49 patients were randomised to the control group (CG) and continued with routine nutritional management. Nutritional status was assessed by weight, anthropometry, and grip strength, with measurements taken at two weekly intervals for 10 weeks. Complications, namely wound infection, chest infection, and antibiotic use were documented. Quality of life (QOL) was assessed using the UK SF-36 questionnaire. RESULTS: Patients in the control group lost a maximum mean (SD) of 5.96 (4.21) kg in weight over a period of eight weeks while patients in group TG lost less weight overall (maximum mean (SD) 3.40 (0.89) kg (p<0.001) occurring at four weeks and progressively regained weight from week 4). Anthropometry, grip strength, and QOL were similarly significantly different between groups (p<0.001). Fewer patients in the treatment group (7/52) required antibiotic prescriptions compared with the control group (15/49). CONCLUSIONS: Nutritional status declined for two months after discharge. Postoperative nutritional supplementation improved nutritional status, QOL, and morbidity in these patients. PMID- 10807892 TI - Do eicosanoids cause colonic dysfunction in experimental E coli O157:H7 (EHEC) infection? AB - BACKGROUND: The pathophysiology of enterohaemorrhagic Escherichia coli (EHEC) infection remains unclear. Eicosanoids have been implicated as pathophysiological mediators in other colitides. AIMS: To determine if prostaglandin E(2) (PGE(2)) and leukotriene B(4) (LTB(4)) contribute to mucosal inflammation and dysfunction in EHEC colitis. METHODS: Ten day old rabbits were infected with EHEC. For five days after infection, mucosal synthesis of PGE(2) and LTB(4) was measured in distal colonic tissue from control and infected animals and (51)Cr-EDTA permeability was assessed in vivo. Myeloperoxidase activity was measured and histological inflammation and damage were assessed at five days in control and infected animals and after treatment of infected animals with the LTB(4) synthesis inhibitor MK-886. In separate experiments, ion transport was measured in Ussing chambers, before and after in vitro addition of the cyclooxygenase inhibitor indomethacin. RESULTS: LTB(4) synthesis was increased from day 2 after infection onwards and PGE(2) synthesis was increased on day 3. Mucosal permeability did not increase until day 5 after infection. MK-886 inhibited colonic LTB(4) production but did not reduce diarrhoea, inflammation, or mucosal damage. Electrolyte transport was not significantly altered on day 3 after infection. However, both Cl secretion and reduced Na absorption found on day 5 were partially reversed by indomethacin. CONCLUSIONS: Tissue synthesis of PGE(2) and LTB(4) did not correlate temporally with EHEC induced inflammation or changes in mucosal permeability and ion transport. Cyclooxygenase inhibition partially reversed ion transport abnormalities but lipoxygenase inhibition did not affect mucosal inflammation or histological damage. We conclude that the contribution of eicosanoids to mucosal injury and dysfunction is more complex than previously suggested. PMID- 10807894 TI - Absorption of calcium and magnesium in patients with intestinal resections treated with medium chain fatty acids. AB - BACKGROUND: Steatorrhoea is associated with increased faecal loss of calcium and magnesium. Medium chain C8-C10 triglycerides (MCTs) improve fat absorption in patients with small bowel resections but the effects on intestinal absorption of divalent cations are not clear. AIM: To assess the effect of dietary replacement of long chain triglycerides (LCTs) with MCTs on calcium and magnesium absorption in patients with small bowel resections. PATIENTS: Nineteen adult patients with a remaining small intestine averaging 171 cm (range 50-300). METHODS: In a crossover design, patients were randomised to two high fat diets (10 MJ/day, 50% as fat) for four days each separated by one day of washout. Diets were prepared in duplicate and were based on either LCT (LCT period) or equal quantities of LCT and MCT (L/MCT period). Metabolic balances were calculated during the last three days of each period. RESULTS: Mean stool volume increased significantly with the L/MCT diet and was 336 ml more than that with the LCT diet (95% confidence interval of mean difference, 26-649 ml). There was no significant change in the net absorption of calcium and magnesium between the two diets. On average, percentage calcium absorption was 8.6% with the LCT diet and 12.5% with the L/MCT diet. Mean percentage magnesium absorption was 5.4% with the LCT diet and 2.9% with the L/MCT diet. CONCLUSIONS: Dietary replacement of 50% long chain triglycerides with medium chain triglycerides in small bowel resected patients increased faecal volume significantly. No changes in the intestinal net absorption of calcium and magnesium were demonstrated. PMID- 10807895 TI - Food induced stimulation of the antisecretory factor can improve symptoms in human inflammatory bowel disease: a study of a concept. AB - BACKGROUND: Antisecretory factor (AF), a 41 kDa cloned and sequenced protein, suppresses intestinal inflammation and hypersecretion in animals. Endogenous AF production can be induced by dietary modifications in several animal species, and this feed has been shown to reduce the incidence of diarrhoeal disease in weaning piglets. The role of AF in intestinal disease in humans is not known. AIMS: To study the effects of hydrothermally processed cereals, optimised for AF induction in animals, added to the diet of patients with longstanding symptoms of inflammatory bowel disease (IBD). PATIENTS: Fifty three patients with IBD (ulcerative colitis and Crohn's disease) were entered into the study, and 50 completed follow up. The experimental group consisted of 16 females (mean age 50 (SEM 5) years) and 10 males (41 (4) years) and the placebo group of 12 women (41 (4) years old) and 12 men (51 (5) years). METHODS: Patients were randomised to receive either hydrothermally processed cereals (active treatment) or the same amount of ordinary cereals (placebo treatment) for four weeks in a double blind study design. Baseline diet and medications remained unchanged. Bowel symptoms, plasma levels of AF, and colonic biopsies were evaluated before and after treatment. RESULTS: The active treatment significantly improved subjective ratings of clinical symptoms and increased plasma AF levels compared with placebo. Plasma lipid levels were unaffected. CONCLUSION: Hydrothermally processed cereals can induce AF production in human IBD. This increase in endogenous AF activity is associated with clinical improvement. Further studies are warranted to clarify the exact role of AF in human intestinal disease. PMID- 10807896 TI - Carbonylation and disassembly of the F-actin cytoskeleton in oxidant induced barrier dysfunction and its prevention by epidermal growth factor and transforming growth factor alpha in a human colonic cell line. AB - BACKGROUND: Intestinal barrier dysfunction concomitant with high levels of reactive oxygen metabolites (ROM) in the inflamed mucosa have been observed in inflammatory bowel disease (IBD). The cytoskeletal network has been suggested to be involved in the regulation of barrier function. Growth factors (epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha)) protect gastrointestinal barrier integrity against a variety of noxious agents. However, the underlying mechanisms of oxidant induced disruption and growth factor mediated protection remain elusive. AIMS: To determine: (1) if oxidation and disassembly of actin (a key cytoskeletal component) plays a major role in ROM induced epithelial monolayer barrier dysfunction; and (2) if growth factor mediated protection involves prevention of theses alterations. METHODS: Caco-2 monolayers were preincubated with EGF, TGF-alpha, or vehicle before incubation with ROM (H(2)O(2) or HOCl). Effects on cell integrity, barrier function, and G- and F-actin (oxidation, disassembly, and assembly) were determined. RESULTS: ROM dose dependently and significantly increased F- and G-actin oxidation (carbonylation), decreased the stable F-actin fraction (index of stability), and increased the monomeric G-actin fraction (index of disassembly). Concomitant with these changes were disruption of the actin cytoskeleton and loss of the monolayer barrier function. In contrast, growth factor pretreatment decreased actin oxidation and enhanced the stable F-actin, while in concert prevented actin disruption and restored normal barrier function of monolayers exposed to ROM. Cytochalasin-D, an inhibitor of actin assembly, not only caused actin disassembly and barrier dysfunction but also abolished the protective action of growth factors. Moreover, an actin stabilising agent, phalloidin, mimicked the protective actions of the growth factors. CONCLUSIONS: Oxidation, disassembly, and instability of the actin cytoskeleton appears to play a key role in the mechanism of oxidant induced loss of intestinal barrier integrity. In contrast, organisation and stabilisation of actin through promotion of its assembly plays a critical role in the mechanism of growth factor mediated protection. PMID- 10807897 TI - Use of (99m)Tc-DISIDA biliary scanning with morphine provocation for the detection of elevated sphincter of Oddi basal pressure. AB - BACKGROUND: Endoscopic biliary manometry is useful in the assessment of patients with types II and III sphincter of Oddi dysfunction, but it is time consuming and invasive. AIM: To investigate the role of (99m)Tc-DISIDA scanning, with and without morphine provocation, as a non-invasive investigation in these patients compared with endoscopic biliary manometry. SUBJECTS AND METHODS: A total of 34 patients with a clinical diagnosis of type II (n = 21) or III (n = 13) sphincter of Oddi dysfunction were studied. Biliary scintigraphy with 100 MBq of (99m)Tc DISIDA was carried out with and without morphine provocation (0.04 mg/kg intravenously) and time/activity curves were compared with the results of subsequent endoscopic biliary manometry. RESULTS: Eighteen (nine type II, nine type III) of the 34 (53%) patients had sphincter of Oddi basal pressures above the upper limit of normal (40 mm Hg). In the standard DISIDA scan without morphine, no significant differences were observed in time to maximal activity (Tmax) or percentage excretion at 45 or 60 minutes between those with normal and those with abnormal biliary manometry. However, following morphine provocation, median percentage excretion at 60 minutes was 4.9% in those with abnormal manometry and 28.2% in the normal manometry group (p = 0.002). Using a cut off value of 15% excretion at 60 minutes, the sensitivity for detecting elevated sphincter of Oddi basal pressure by the morphine augmented DISIDA scan was 83% and specificity was 81%. Also, 14 of the 18 patients with abnormal manometry complained of biliary-type pain after morphine infusion compared with only two of 16 patients in the normal manometry group (p = 0.001). CONCLUSIONS: (99m)Tc DISIDA with morphine provocation is a useful non-invasive investigation for types II and III sphincter of Oddi dysfunction to detect those with elevated sphincter basal pressures who may respond to endoscopic sphincterotomy. PMID- 10807898 TI - Cancer of the ampulla of Vater: chromosome 17p allelic loss is associated with poor prognosis. AB - BACKGROUND: Cancer of the ampulla of Vater kills 60% of affected patients. Local spread of the tumour (T stage) is the only reliable prognostic factor. Nevertheless, any cancer stage includes long term survivors and patients dying from the disease. The molecular anomalies involved in this process have the potential to serve as additional prognostic markers. AIM: To evaluate if allelic losses (LOH) of chromosomes 17p and 18q may be of prognostic value in multivariate survival analysis. METHODS: We examined 53 ampullary cancers for chromosome 17p and 18q LOH using microsatellite markers and DNA from paraffin embedded tumours. All patients were treated by surgery alone (pancreaticoduodenectomy). Multivariate survival analysis included age, sex, tumour size, macroscopic appearance, grade of differentiation, T stage, lymph node metastasis, and chromosome 17p and 18q status. RESULTS: Chromosome 17p and 18q LOH were detected in 28 (53%) and 18 (34%) cancers, respectively. Multivariate survival analysis indicated chromosome 17p status as an independent prognostic factor together with T stage. The five year survival for chromosome 17p retention and 17p loss was 80% and 7%, respectively. The risk of death from cancer within the five year follow up period for patients with cancers harbouring chromosome 17p LOH was 11 times higher than that of patients with cancers retaining chromosome 17p (p<0.0001), regardless of the tumour stage at diagnosis. CONCLUSIONS: Chromosome 17p status is an independent prognostic factor among ampullary cancers at the same stage. The combined use of T stage and chromosome 17p status may help in deciding whether ampullary cancer patients require additional therapy other than surgery alone. PMID- 10807899 TI - Post-feeding hyperammonaemia in patients with transjugular intrahepatic portosystemic shunt and liver cirrhosis: role of small intestinal ammonia release and route of nutrient administration. AB - BACKGROUND: Hyperammonaemia is a pathogenetic factor for hepatic encephalopathy that may be augmented after a transjugular intrahepatic portosystemic shunt (TIPS). Experimental data suggest that hyperammonaemia may be caused to a large extent by metabolism of small intestinal enterocytes rather than colonic bacteria. AIMS: To evaluate if ammonia release and glutamine metabolism by small intestinal mucosa contribute to hyperammonaemia in vivo in patients with liver cirrhosis. METHODS: Using TIPS to examine mesenteric venous blood, we measured mesenteric venous-arterial concentration differences in ammonia and glutamine in patients with liver cirrhosis before, during, and after enteral (n = 8) or parenteral (n = 8) isonitrogenous infusion of a glutamine containing amino acid solution. RESULTS: During enteral nutrient infusion, ammonia release increased rapidly compared with the post-absorptive state (65 (58-73) v. 107 (95-119) micromol/l after 15 min; mean (95% confidence interval)) in contrast with parenteral infusion (50 (41-59) v. 62 (47-77) micromol/l). This resulted in a higher portal ammonia load (29 (21-36) v. 14 (8-21) mmol/l/240 minutes) and a higher degree of systemic hyperammonaemia (14 (11-17) v. 9 (6-12) mmol/l/240 minutes) during enteral than parenteral infusion. The mesenteric venous-arterial concentration difference in glutamine changed from net uptake to release at the end of the enteral infusion period (-100 (-58 to -141) v. 31 (-47-110) micromol/l) with no change during parenteral nutrition. CONCLUSIONS: These data suggest that small intestinal metabolism contributes to post-feeding hyperammonaemia in patients with cirrhosis. When artificial nutrition is required, parenteral nutrition may be superior to enteral nutrition in patients with portosystemic shunting because of the lower degree of systemic hyperammonaemia. PMID- 10807900 TI - CO(2) wedged hepatic venography in the evaluation of portal hypertension. AB - BACKGROUND/AIMS/METHODS: During hepatic vein catheterisation, in addition to measurement of hepatic venous pressure gradient (HVPG), iodine wedged retrograde portography can be easily obtained. However, it rarely allows correct visualisation of the portal vein. Recently, CO(2) has been suggested to allow better angiographic demonstration of the portal vein than iodine. In this study we investigated the efficacy of CO(2) compared with iodinated contrast medium for portal vein imaging and its role in the evaluation of portal hypertension in a series of 100 patients undergoing hepatic vein catheterisation, 71 of whom had liver cirrhosis. RESULTS: In the overall series, CO(2) venography was markedly superior to iodine, allowing correct visualisation of the different segments of the portal venous system. In addition, CO(2), but not iodine, visualised portal systemic collaterals in 34 patients. In cirrhosis, non-visualisation of the portal vein on CO(2) venography occurred in 11 cases; four had portal vein thrombosis and five had communications between different hepatic veins. Among non cirrhotics, lack of portal vein visualisation had a 90% sensitivity, 88% specificity, 94% negative predictive value, and 83% positive predictive value in the diagnosis of pre-sinusoidal portal hypertension. CONCLUSIONS: Visualisation of the venous portal system by CO(2) venography is markedly superior to iodine. The use of CO(2) wedged portography is a useful and safe complementary procedure during hepatic vein catheterisation which may help to detect portal thrombosis. Also, lack of demonstration of the portal vein in non-cirrhotic patients strongly suggests the presence of pre-sinusoidal portal hypertension. PMID- 10807901 TI - Neuromyogenic properties of the internal anal sphincter: therapeutic rationale for anal fissures. AB - Lateral sphincterotomy diminishes internal anal sphincter hypertonia and thereby reduces anal canal pressure. This improves anal mucosal blood flow and promotes the healing of anal fissures. However, sphincterotomy can be associated with long term disturbances of sphincter function. The optimal treatment for an anal fissure is to induce a temporary reduction of anal canal resting pressure to allow healing of the fissure without permanently disrupting normal sphincter function. Broader understanding of the intrinsic mechanisms controlling smooth muscle contraction has allowed pharmacological manipulation of anal sphincter tone. We performed an initial Medline literature search to identify all articles concerning "internal anal sphincter" and "anal fissures". This review is based on these articles and on additional publications obtained by manual cross referencing. Internal anal smooth muscle relaxation can be inhibited by stimulation of non-adrenergic non-cholinergic enteric neurones, parasympathetic muscarinic receptors, or sympathetic beta adrenoceptors, and by inhibition of calcium entry into the cell. Sphincter contraction depends on an increase in cytoplasmic calcium and is enhanced by sympathetic adrenergic stimulation. Currently, the most commonly used pharmacological agent in the treatment of anal fissures is topical glyceryl trinitrate, a nitric oxide donor. Alternative agents that exhibit a similar effect via membrane Ca2+ channels, muscarinic receptors, and alpha or beta adrenoceptors are also likely to have a therapeutic potential in treating anal fissures. PMID- 10807902 TI - Stem cell transplantation for inflammatory bowel disease: practical and ethical issues. PMID- 10807903 TI - Dopamine D5 receptor agonist high affinity and constitutive activity profile conferred by carboxyl-terminal tail sequence. AB - The mammalian dopamine D1-like receptor gene family is comprised of two members, termed D1/D1A and D5/D1B. In an attempt to define the role of the carboxyl terminal (CT) tail in the expression of D5 subtype-specific pharmacological and constitutive activity profiles, we examined a series of D5 receptor chimeras in which only the CT tail was swapped with corresponding sequences encoding human/vertebrate D1-like receptors. D5/D1(CT) or D5/D1D(CT) tail substitution mutants displayed a rank order of potency and agonist affinities virtually mimicking wild-type (wt) D1 receptors, as indexed by both ligand binding and dopamine-stimulated cAMP accumulation assays, and, similar to wt D1 receptors, did not exhibit receptor constitutive activity or responsiveness to inverse agonists. D1/D5(CT) or D1/D1D(CT) tail receptor mutants displayed agonist pharmacological and functional characteristics not significantly different from parental D1 or mutant D5/D1(CT) and D5/D1D(CT) receptors. The affinities for numerous antagonists remained essentially unchanged for all receptor chimeras relative to parental wt receptors. A series of stepwise D5-CT-tail truncation/deletion mutants identified the region encoded by amino acids 438-448 and particularly Gln(439), as necessary and sufficient for the full expression of high affinity agonist and functional D5 receptor characteristics. Site-directed mutagenesis of the highly conserved D5/D1B receptor residue Gln(439)-(Ala/Ile), converts the full-length D5 receptor to one displaying "super" D5 characteristics with expressed affinities for discriminating agonists approximately 4- to 5-fold higher than wt D5 but without any concomitant increases of agonist-independent basal cAMP accumulation or intrinsic activity. Taken together, these data suggest that, in addition to other well characterized receptor domains, the agonist pharmacological and functional signature of the D5/D1B receptor is modulated by sequence-specific motifs within the CT tail and that one conserved amino acid in this region can further regulate D5 agonist high affinity binding interactions independent of receptor constitutive activity. PMID- 10807904 TI - The tumor necrosis factor-related apoptosis-inducing ligand receptors TRAIL-R1 and TRAIL-R2 have distinct cross-linking requirements for initiation of apoptosis and are non-redundant in JNK activation. AB - Overexpression of the tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptors, TRAIL-R1 and TRAIL-R2, induces apoptosis and activation of NF-kappaB in cultured cells. In this study, we have demonstrated differential signaling capacities by both receptors using either epitope-tagged soluble TRAIL (sTRAIL) or sTRAIL that was cross-linked with a monoclonal antibody. Interestingly, sTRAIL was sufficient for induction of apoptosis only in cell lines that were killed by agonistic TRAIL-R1- and TRAIL-R2-specific IgG preparations. Moreover, in these cell lines interleukin-6 secretion and NF-kappaB activation were induced by cross-linked or non-cross-linked anti-TRAIL, as well as by both receptor-specific IgGs. However, cross-linking of sTRAIL was required for induction of apoptosis in cell lines that only responded to the agonistic anti-TRAIL-R2-IgG. Interestingly, activation of c-Jun N-terminal kinase (JNK) was only observed in response to either cross-linked sTRAIL or anti-TRAIL-R2-IgG even in cell lines where both receptors were capable of signaling apoptosis and NF kappaB activation. Taken together, our data suggest that TRAIL-R1 responds to either cross-linked or non-cross-linked sTRAIL which signals NF-kappaB activation and apoptosis, whereas TRAIL-R2 signals NF-kappaB activation, apoptosis, and JNK activation only in response to cross-linked TRAIL. PMID- 10807905 TI - The human thioesterase II protein binds to a site on HIV-1 Nef critical for CD4 down-regulation. AB - A HIV-1 Nef affinity column was used to purify a 35-kDa Nef-interacting protein from T-cell lysates. The 35-kDa protein was identified by peptide microsequence analysis as the human thioesterase II (hTE) enzyme, an enzyme previously identified in a yeast two-hybrid screen as a potential Nef-interacting protein. Immunofluorescence studies showed that hTE localizes to peroxisomes and that coexpression of Nef and hTE leads to relocalization of Nef to peroxisomes. Interaction of Nef and hTE was abolished by point mutations in Nef at residues Asp(108), Leu(112), Phe(121), Pro(122), and Asp(123). All of these mutations also abrogated the ability of Nef to down-regulate CD4 from the surface of HIV infected cells. Based on the x-ray and NMR structures of Nef, these residues define a surface on Nef critical for CD4 down-regulation. A subset of these mutations also affected the ability of Nef to down-regulate major histocompatibility complex class I. These results, taken together with previous studies, identify a region on Nef critical for most of its known functions. However, not all Nef alleles bind to hTE with high affinity, so the role of hTE during HIV infection remains uncertain. PMID- 10807906 TI - Two glycosylase/abasic lyases from Neisseria mucosa that initiate DNA repair at sites of UV-induced photoproducts. AB - Diverse organisms ranging from Escherichia coli to humans contain a variety of DNA repair proteins that function in the removal of damage caused by shortwave UV light. This study reports the identification, purification, and biochemical characterization of two DNA glycosylases with associated abasic lyase activity from Neisseria mucosa. These enzymes, pyrimidine dimer glycosylase I and II (Nmu pdg I and Nmu-pdg II), were purified 30,000- and 10,000-fold, respectively. SDS polyacrylamide gel electrophoresis analysis indicated that Nmu-pdg I is approximately 30 kDa, whereas Nmu-pdg II is approximately 19 kDa. The N-terminal amino acid sequence of Nmu-pdg II exhibits 64 and 66% identity with E. coli and Hemophilus parainfluenzae endonuclease III, respectively. Both Nmu-pdg I and Nmu pdg II were found to have broad substrate specificities, as evidenced by their ability to incise DNA containing many types of UV and some types of oxidative damage. Consistent with other glycosylase/abasic lyases, the existence of a covalent enzyme-DNA complex could be demonstrated for both Nmu-pdg I and II when reactions were carried out in the presence of sodium borohydride. These data indicate the involvement of an amino group in the catalytic reaction mechanism of both enzymes. PMID- 10807907 TI - Down-regulation of insulin signaling by protein-tyrosine phosphatase 1B is mediated by an N-terminal binding region. AB - Protein-tyrosine phosphatases (PTPs) play a major role in regulating insulin signaling. Among the PTPs that regulate this signaling pathway, PTP1B plays an especially prominent role. PTP1B inhibits insulin signaling and has previously been shown to bind to the activated insulin receptor (IR), but neither the mechanism nor the physiological importance of such binding have been established. Here, we show that a previously undefined region in the N-terminal, catalytic half of PTP1B contributes to IR binding. Point mutations within this region of PTP1B disrupt IR binding but do not affect the catalytic activity of this phosphatase. This binding-defective mutant of PTP1B does not efficiently dephosphorylate the IR in cells, nor does it effectively inhibit IR signaling. These results suggest that PTP1B targets the IR through a novel binding element and that binding is required for the physiological effects of PTP1B on IR signal transduction. PMID- 10807908 TI - A role of the Ca2+/Mg2+-dependent endonuclease in apoptosis and its inhibition by Poly(ADP-ribose) polymerase. AB - Apoptosis is characterized by various cell morphological and biochemical features, one of which is the internucleosomal degradation of genomic DNA. The role of the human chromatin-bound Ca(2+)- and Mg(2+)-dependent endonuclease (CME) DNAS1L3 and its inhibition by poly(ADP-ribosyl)ation in the DNA degradation that accompanies apoptosis was investigated. The nuclear localization of this endonuclease is the unique feature that distinguishes it from other suggested apoptotic nucleases. Purified recombinant DNAS1L3 was shown to cleave nuclear DNA into both high molecular weight and oligonucleosomal fragments in vitro. Furthermore, exposure of mouse skin fibroblasts expressing DNAS1L3 to inducers of apoptosis resulted in oligonucleosomal DNA fragmentation, an effect not observed in cells not expressing this CME, as well as in a decrease in cell viability greater than that apparent in the control cells. Recombinant DNAS1L3 was modified by recombinant human poly(ADP-ribose) polymerase (PARP) in vitro, resulting in a loss of nuclease activity. The DNAS1L3 protein also underwent poly(ADP ribosyl)ation in transfected mouse skin fibroblasts in response to inducers of apoptosis. The cleavage and inactivation of PARP by a caspase-3-like enzyme late in apoptosis were associated with a decrease in the extent of DNAS1L3 poly(ADP ribosyl)ation, which likely releases DNAS1L3 from inhibition and allows it to catalyze the degradation of genomic DNA. PMID- 10807909 TI - Phorbol esters and cytokines regulate the expression of the NEMO-related protein, a molecule involved in a NF-kappa B-independent pathway. AB - The NF-kappaB signaling pathway plays a crucial role in the immune, inflammatory, and apoptotic responses. Recently, we identified the NF-kappaB Essential Modulator (NEMO) as an essential component of this pathway. NEMO is a structural and regulatory subunit of the high molecular kinase complex (IKK) responsible for the phosphorylation of NF-kappaB inhibitors. Data base searching led to the isolation of a cDNA encoding a protein we called NRP (NEMO-related protein), which shows a strong homology to NEMO. Here we show that NRP is present in a novel high molecular weight complex, that contains none of the known members of the IKK complex. Consistently, we could not observe any effect of NRP on NF kappaB signaling. Nonetheless, we could demonstrate that treatment with phorbol esters induces NRP phosphorylation and decreases its half-life. This phosphorylation event could only be inhibited by K-252a and stauroporin. We also show that de novo expression of NRP can be induced by interferon and tumor necrosis factor alpha and that these two stimuli have a synergistic effect on NRP expression. In addition, we observed that endogenous NRP is associated with the Golgi apparatus. Analogous to NEMO, we find that NRP is associated in a complex with two kinases, suggesting that NRP could play a similar role in another signaling pathway. PMID- 10807911 TI - Prolactin-induced cell proliferation in PC12 cells depends on JNK but not ERK activation. AB - The effects of pituitary and extrapituitary prolactin include cellular proliferation and differentiation. PC12 cells was used as a model to delineate respective signaling of prolactin. Prolactin acted as a mitogen for undifferentiated PC12 cells, as measured by significant increases in bromodeoxyuridine incorporation and in cell numbers, with an efficacy equal to epidermal growth factor. Both the long and short form of the prolactin receptor was expressed, yet only the long isoform was tyrosine-phosphorylated upon agonist binding. Functional prolactin receptor signaling was further demonstrated in the activation of JAK2 and phosphorylation activation of the transcription factors Stat1, -3, and -5a. Surprisingly, prolactin stimulated a sustained activation of Raf-B, without activation of the MAP kinases ERK1 or -2. Instead, in solid phase kinase assays using a glutathione S-transferase-c-Jun fusion protein (amino acids 1-79) as the substrate, a significant activation of the mitogen-activated protein Janus kinase (c-Jun N-terminal kinase; JNK) was observed. The prolactin-induced activation of JNK was prolonged and accompanied by a significant increase in c Jun mRNA abundance and c-Jun protein synthesis. Moreover, analysis of bromodeoxyuridine incorporation at the single cell level revealed that epidermal growth factor-dependent incorporation was inhibited by PD98059 and independent of SB203580, whereas prolactin-induced incorporation was ERK and mitogen-activated protein kinase p38 independent but was abolished with JNK inhibition by 30 microm SB203580. Our studies suggest that prolactin may have a role in the growth of PC12 cells, where it stimulates concurrent mitogenic and differentiation promoting signaling pathways. PMID- 10807912 TI - Degradation of the epstein-barr virus latent membrane protein 1 (LMP1) by the ubiquitin-proteasome pathway. Targeting via ubiquitination of the N-terminal residue. AB - The latent membrane protein 1 (LMP1) of the Epstein-Barr virus is a constitutively active receptor essential for B lymphocyte transformation by the Epstein-Barr virus. It is a short-lived protein, but the proteolytic pathway involved in its degradation is not known. The ubiquitin pathway is a major system for specific protein degradation in eukaryotes. Most plasma membrane substrates of the pathway are internalized upon ubiquitination and delivered for degradation in the lysosome/vacuole. Here we show that LMP1 is a substrate of the ubiquitin pathway and is ubiquitinated both in vitro and in vivo. However, in contrast to other plasma membrane substrates of the ubiquitin system, it is degraded mostly by the proteasome and not by lysosomes. Degradation is independent of the single Lys residue of the protein; a lysine-less mutant LMP1 is degraded in a ubiquitin- and proteasome-dependent manner similar to the wild type protein. Degradation of both wild type and lysine-less protein is sensitive to fusion of a Myc tag to the N terminus of LMP1. In addition, deletion of as few as 12 N-terminal amino acid residues stabilizes the protein. These findings suggest that the first event in LMP1 degradation is attachment of ubiquitin to the N-terminal residue of the protein. We present evidence suggesting that phosphorylation is also required for degradation of LMP1. PMID- 10807913 TI - Sequences within the DNA cross-linking patch of sigma 54 involved in promoter recognition, sigma isomerization, and open complex formation. AB - The bacterial RNA polymerase holoenzyme containing the final sigma(54) subunit functions in enhancer-dependent transcription. Mutagenesis has been used to probe the function of a sequence in the final sigma(54) DNA binding domain that includes residues that cross-link to promoter DNA. Several activities of the final sigma and holoenzyme are shown to depend on the cross-linking patch. The patch contributes to promoter binding by final sigma(54), and holoenzyme and is involved in activator-dependent final sigma isomerization. As part of the final sigma(54)-holoenzyme, some residues in the patch limit basal transcription. Other cross-linking patch sequences appear to limit activator-dependent open complex formation. Deletion of 19 residues adjacent to the cross-linking patch resulted in a holoenzyme unable to respond to activator but capable of activator independent (bypass) transcription in vitro. Overall results are consistent with the cross-linking patch directing interactions to the -12 promoter region to set basal and activated levels of transcription. PMID- 10807914 TI - Relative spatial position of a snake neurotoxin and the reduced disulfide bond alpha (Cys192-Cys193) at the alpha gamma interface of the nicotinic acetylcholine receptor. AB - We determined the distances separating five functionally important residues (Gln(10), Lys(27), Trp(29), Arg(33), and Lys(47)) of a three-fingered snake neurotoxin from the reduced disulfide bond alpha(Cys(192)-Cys(193)) located at the alphagamma interface of the Torpedo nicotinic acetylcholine receptor. Each toxin position was substituted individually for a cysteine, which was then linked to a maleimido moiety through three different spacers, varying in length from 10 to 22 A. We estimated the coupling efficiency between the 15 toxin derivatives and the reduced cystine alpha(192-193) by gel densitometry of Coomassie Blue stained gels. A nearly quantitative coupling was observed between alphaCys(192) and/or alphaCys(193) and all probes introduced at the tip of the first (position 10) and second (position 33) loops of Naja nigricollis alpha-neurotoxin. These data sufficed to locate the reactive thiolate in a "croissant-shaped" volume comprised between the first two loops of the toxin. The volume was further restrained by taking into account the absence or partial coupling of the other derivatives. Altogether, the data suggest that alphaCys(192) and/or alphaCys(193), at the alphagamma interface of a muscular-type acetylcholine receptor, is (are) located in a volume located between 11.5 and 15.5 A from the alpha-carbons at positions 10 and 33 of the toxin, under the tip of the toxin first loop and close to the second one. PMID- 10807915 TI - Regulated targeting of a protein kinase into an intact flagellum. An aurora/Ipl1p like protein kinase translocates from the cell body into the flagella during gamete activation in chlamydomonas. AB - In the green alga Chlamydomonas reinhardtii flagellar adhesion between gametes of opposite mating types leads to rapid cellular changes, events collectively termed gamete activation, that prepare the gametes for cell-cell fusion. As is true for gametes of most organisms, the cellular and molecular mechanisms that underlie gamete activation are poorly understood. Here we report on the regulated movement of a newly identified protein kinase, Chlamydomonas aurora/Ipl1p-like protein kinase (CALK), from the cell body to the flagella during gamete activation. CALK encodes a protein of 769 amino acids and is the newest member of the aurora/Ipl1p protein kinase family. Immunoblotting with an anti-CALK antibody showed that CALK was present as a 78/80-kDa doublet in vegetative cells and unactivated gametes of both mating types and was localized primarily in cell bodies. In cells undergoing fertilization, the 78-kDa CALK was rapidly targeted to the flagella, and within 5 min after mixing gametes of opposite mating types, the level of CALK in the flagella began to approach levels normally found in the cell body. Protein synthesis was not required for targeting, indicating that the translocated CALK and the cellular molecules required for its movement are present in unactivated gametes. CALK was also translocated to the flagella during flagellar adhesion of nonfusing mutant gametes, demonstrating that cell fusion was not required for movement. Finally, the requirement for flagellar adhesion could be bypassed; incubation of cells of a single mating type in dibutyryl cAMP led to CALK translocation to flagella in gametes but not vegetative cells. These experiments document a new event in gamete activation in Chlamydomonas and reveal the existence of a mechanism for regulated translocation of molecules into an intact flagellum. PMID- 10807916 TI - MEKK4 mediates differentiation in response to retinoic acid via activation of c Jun N-terminal kinase in rat embryonal carcinoma P19 cells. AB - Differentiation of P19 embryonal carcinoma cells in response to the morphogen retinoic acid is regulated by Galpha(12/13) and is associated with activation of c-Jun N-terminal kinase. The role of MEKK1 and MEKK4 upstream of the c-Jun N terminal kinase was investigated in P19 cells. P19 clones stably expressing constitutively active and dominant negative mutants of MEKK1 and MEKK4 were created and characterized. Expression of the constitutively active form of either MEKK1 or MEKK4 mimicked the action of retinoic acid, inducing these embryonal carcinoma cells to primitive endoderm. Expression of the dominant negative form of MEKK1 had no influence on the ability of retinoic acid to induce either JNK activation or primitive endoderm formation in P19 stem cells. Expression of the dominant negative form of MEKK4, in contrast, effectively blocks both morphogen induced activation of JNK and cellular differentiation. These data identify MEKK4 as upstream of c-Jun N-terminal kinase in the pathway mediating differentiation of P19 stem cells to primitive endoderm. PMID- 10807917 TI - Complexes between protein export chaperone SecB and SecA. Evidence for separate sites on SecA providing binding energy and regulatory interactions. AB - During localization to the periplasmic space or to the outer membrane of Escherichia coli some proteins are dependent on binding to the cytosolic chaperone SecB, which in turn is targeted to the membrane by specific interaction with SecA, a peripheral component of the translocase. Five variant forms of SecB, previously demonstrated to be defective in mediating export in vivo (Gannon, P. M., and Kumamoto, C. A. (1993) J. Biol. Chem. 268, 1590-1595; Kimsey, H. K., Dagarag, M. D., and Kumamoto, C. A. (1995) J. Biol. Chem. 270, 22831-22835) were investigated with respect to their ability to bind SecA both in solution and at the membrane translocase. We present evidence that at least two regions of SecA are involved in the formation of active complexes with SecB. The variant forms of SecB were all capable of interacting with SecA in solution to form complexes with stability similar to that of complexes between SecA and wild-type SecB. However, the variant forms were defective in interaction with a separate region of SecA, which was shown to trigger a change that was correlated to activation of the complex. The region of SecA involved in activation of the complexes was defined as the extreme carboxyl-terminal 21 aminoacyl residues. PMID- 10807918 TI - Transactivation of the EGF receptor mediates IGF-1-stimulated shc phosphorylation and ERK1/2 activation in COS-7 cells. AB - The receptor for insulin-like growth factor 1 (IGF-1) mediates multiple cellular responses, including stimulation of both proliferative and anti-apoptotic pathways. We have examined the role of cross talk between the IGF-1 receptor (IGF 1R) and the epidermal growth factor receptor (EGFR) in mediating responses to IGF 1. In COS-7 cells, IGF-1 stimulation causes tyrosine phosphorylation of the IGF 1R beta subunit, the EGFR, insulin receptor substrate-1 (IRS-1), and the Shc adapter protein. Shc immunoprecipitates performed after IGF-1 stimulation contain coprecipitated EGFR, suggesting that IGF-1R activation induces the assembly of EGFR.Shc complexes. Tyrphostin AG1478, an inhibitor of the EGFR kinase, markedly attenuates IGF-1-stimulated phosphorylation of EGFR, Shc, and ERK1/2 but has no effect on phosphorylation of IGF-1R, IRS-1, and protein kinase B (Akt). Cross talk between IGF-1 and EGF receptors is mediated through an autocrine mechanism involving matrix metalloprotease-dependent release of heparin-binding EGF (HB EGF), because IGF-1-mediated ERK activation is inhibited both by [Glu(52)]Diphtheria toxin, a specific inhibitor of HB-EGF, and the metalloprotease inhibitor 1,10-phenanthroline. These data demonstrate that IGF-1 stimulation of the IRS-1/PI3K/Akt pathway and the EGFR/Shc/ERK1/2 pathway occurs by distinct mechanisms and suggest that IGF-1-mediated "transactivation" of EGFR accounts for the majority of IGF-1-stimulated Shc phosphorylation and subsequent activation of the ERK cascade. PMID- 10807919 TI - Mechanism of heavy metal ion activation of phytochelatin (PC) synthase: blocked thiols are sufficient for PC synthase-catalyzed transpeptidation of glutathione and related thiol peptides. AB - The dependence of phytochelatin synthase (gamma-glutamylcysteine dipeptidyltranspeptidase (PCS), EC ) on heavy metals for activity has invariably been interpreted in terms of direct metal binding to the enzyme. Here we show, through analyses of immunopurified, recombinant PCS1 from Arabidopsis thaliana (AtPCS1), that free metal ions are not essential for catalysis. Although AtPCS1 appears to be primarily activated posttranslationally in the intact plant and purified AtPCS1 is able to bind heavy metals directly, metal binding per se is not responsible for catalytic activation. As exemplified by Cd(2+)- and Zn(2+) dependent AtPCS1-mediated catalysis, the kinetics of PC synthesis approximate a substituted enzyme mechanism in which micromolar heavy metal glutathione thiolate (e.g. Cd.GS(2) or Zn.GS(2)) and free glutathione act as gamma-Glu-Cys acceptor and donor. Further, as demonstrated by the facility of AtPCS1 for the net synthesis of S-alkyl-PCs from S-alkylglutathiones with biphasic kinetics, consistent with the sufficiency of S-alkylglutathiones as both gamma-Glu-Cys donors and acceptors in media devoid of metals, even heavy metal thiolates are dispensable. It is concluded that the dependence of AtPCS1 on the provision of heavy metal ions for activity in media containing glutathione and other thiol peptides is a reflection of this enzyme's requirement for glutathione-like peptides containing blocked thiol groups for activity. PMID- 10807920 TI - Induction of smooth muscle alpha-actin in vascular smooth muscle cells by arginine vasopressin is mediated by c-Jun amino-terminal kinases and p38 mitogen activated protein kinase. AB - Exposure of vascular smooth muscle cells to arginine vasopressin (AVP) increases smooth muscle alpha-actin (SM-alpha-actin) expression through activation of the SM- alpha-actin promoter. The goal of this study was to determine the role of the mitogen-activated protein kinase (MAP kinase) family in regulation of SM-alpha actin expression. AVP activated all three MAP kinase family members: ERKs, JNKs, and p38 MAP kinase. Inhibition of JNKs or p38 decreased AVP-stimulated SM-alpha actin promoter activity, whereas inhibition of ERKs had no effect. A 150-base pair region of the promoter containing two CArG boxes was sufficient to mediate regulation by vasoconstrictors. Mutations in either CArG box decreased AVP stimulated promoter activity. Electrophoretic mobility shift assays using oligonucleotides corresponding to either CArG box resulted in a complex of similar mobility whose intensity was increased by AVP. Antibodies against serum response factor (SRF) completely super-shifted this complex, indicating that SRF binds to both CArG boxes. Overexpression of SRF increased basal promoter activity, but activity was still stimulated by AVP. AVP stimulation rapidly increased SRF phosphorylation. These data indicate that both JNKs and p38 participate in regulation of SM- alpha-actin expression. SRF, which binds to two critical CArG boxes in the promoter, represents a potential target of these kinases. PMID- 10807921 TI - The transmembrane domains of hepatitis C virus envelope glycoproteins E1 and E2 play a major role in heterodimerization. AB - Oligomerization of viral envelope proteins is essential to control virus assembly and fusion. The transmembrane domains (TMDs) of hepatitis C virus envelope glycoproteins E1 and E2 have been shown to play multiple functions during the biogenesis of E1E2 heterodimer. This makes them very unique among known transmembrane sequences. In this report, we used alanine scanning insertion mutagenesis in the TMDs of E1 and E2 to examine their role in the assembly of E1E2 heterodimer. Alanine insertion within the center of the TMDs of E1 or E2 or in the N-terminal part of the TMD of E1 dramatically reduced heterodimerization, demonstrating the essential role played by these domains in the assembly of hepatitis C virus envelope glycoproteins. To better understand the alanine scanning data obtained for the TMD of E1 which contains GXXXG motifs, we analyzed by circular dichroism and nuclear magnetic resonance the three-dimensional structure of the E1-(350-370) peptide encompassing the N-terminal sequence of the TMD of E1 involved in heterodimerization. Alanine scanning results and the three dimensional molecular model we obtained provide the first framework for a molecular level understanding of the mechanism of hepatitis C virus envelope glycoprotein heterodimerization. PMID- 10807922 TI - Three-dimensional structure of Chlamydomonas reinhardtii and Synechococcus elongatus photosystem II complexes allows for comparison of their oxygen-evolving complex organization. AB - Electron microscopy and single-particle analyses have been carried out on negatively stained photosystem II (PSII) complexes isolated from the green alga Chlamydomonas reinhardtii and the thermophilic cyanobacterium Synechococcus elongatus. The analyses have yielded three-dimensional structures at 30-A resolution. Biochemical analysis of the C. reinhardtii particle suggested it to be very similar to the light-harvesting complex II (LHCII).PSII supercomplex of spinach, a conclusion borne out by its three-dimensional structure. Not only was the C. reinhardtii LHCII.PSII supercomplex dimeric and of comparable size and shape to that of spinach, but the structural features for the extrinsic OEC subunits bound to the lumenal surface were also similar thus allowing identification of the PsbO, PsbP, and PsbQ OEC proteins. The particle isolated from S. elongatus was also dimeric and retained its OEC proteins, PsbO, PsbU, and PsbV (cytochrome c(550)), which were again visualized as protrusions on the lumenal surface of the complex. The overall size and shape of the cyanobacterial particle was similar to that of a PSII dimeric core complex isolated from spinach for which higher resolution structural data are known from electron crystallography. By building the higher resolution structural model into the projection maps it has been possible to relate the positioning of the OEC proteins of C. reinhardtii and S. elongatus with the underlying transmembrane helices of other major intrinsic subunits of the core complex, D1, D2, CP47, and CP43 proteins. It is concluded that the PsbO protein is located over the CP47 and D2 side of the reaction center core complex, whereas the PsbP/PsbQ and PsbV/PsbU are positioned over the lumenal surface of the N-terminal region of the D1 protein. However, the mass attributed to PsbV/PsbU seems to bridge across to the PsbO, whereas the PsbP/PsbQ proteins protrude out more from the lumenal surface. Nevertheless, within the resolution and quality of the data, the relative positions of the center of masses for OEC proteins of C. reinhardtii and S. elongatus are similar and consistent with those determined previously for the OEC proteins of spinach. PMID- 10807923 TI - Interaction of inhibitor-2 with the catalytic subunit of type 1 protein phosphatase. Identification of a sequence analogous to the consensus type 1 protein phosphatase-binding motif. AB - Inhibitor-2 (I-2) is the regulatory subunit of a cytosolic type 1 Ser/Thr protein phosphatase (PP1) and potently inhibits the activity of the free catalytic subunit (CS1). Previous work from the laboratory had proposed that the interaction of I-2 with CS1 involved multiple sites (Park, I. K., and DePaoli Roach, A. A. (1994) J. Biol. Chem. 269, 28919-28928). The present study refines the earlier analysis and arrives at a more detailed model for the interaction between I-2 and CS1. Although the NH(2)-terminal I-2 regions containing residues 1-35 and 1-64 have no inhibitory activity on their own, they increase the IC(50) for I-2 by approximately 30-fold, indicating the presence of a CS1-interacting site. Based on several experimental approaches, we have also identified the sequence Lys(144)-Leu-His-Tyr(147) as a second site of interaction that corresponds to the RVXF motif present in many CS1-binding proteins. The peptide I 2(135-151) significantly increases the IC(50) for I-2 and attenuates CS1 inhibition. Replacement of Leu and Tyr with Ala abolishes the ability to counteract inhibition by I-2. The I-2(135-151) peptide, but not I-2(1-35), also antagonizes inhibition of CS1 by DARPP-32 in a pattern similar to that of I-2. Furthermore, a peptide derived from the glycogen-binding subunit, R(GL)/G(M)(61 80), which contains a consensus CS1-binding motif, completely counteracts CS1 inhibition by I-2 and DARPP-32. The NH(2)-terminal 35 residues of I-2 bind to CS1 at a site that is specific for I-2, whereas the KLHY sequence interacts with CS1 at a site shared with other interacting proteins. Other results suggest the presence of yet more sites of interaction. A model is presented in which multiple "anchoring interactions" serve to position a segment of I-2 such that it sterically occludes the catalytic pocket but need not make high affinity contacts itself. PMID- 10807924 TI - Assignment of functional amino acids around the active site of human DNA topoisomerase IIalpha. AB - An expression library for active site mutants of human topoisomerase IIalpha (TOP2alpha) was constructed by replacing the sequence encoding residues 793-808 with a randomized oligonucleotide cassette. This plasmid library was transformed into a temperature-sensitive yeast strain (top2-1), and viable transformants were selected at the restrictive temperature. Among the active TOP2alpha mutants, no substitution was allowed at Tyr(805), the 5' anchor of the cleaved DNA, and only conservative substitutions were allowed at Leu(794), Asp(797), Ala(801), and Arg(804). Thus, these 5 residues are critical for human TOP2alpha activity, and the remaining mutagenized residues are less critical for function. Using the x ray crystal structure of yeast TOP2 as a structural model, it can be deduced that these 5 functionally important residues lie in a plane. One of the possible functions of this plane may be that it interacts with the DNA substrate upon catalysis. The side chains of Ser(803) and Lys(798), which confer drug resistance, lie adjacent to this plane. PMID- 10807925 TI - A mutant of Tetrahymena telomerase reverse transcriptase with increased processivity. AB - The protein catalytic subunit of telomerase (TERT) is a reverse transcriptase (RT) that utilizes an internal RNA molecule as a template for the extension of chromosomal DNA ends. In all retroviral RTs there is a conserved tyrosine two amino acids preceding the catalytic aspartic acids in motif C, a motif that is critical for catalysis. In TERTs, however, this position is a leucine, valine, or phenylalanine. We developed and characterized a robust in vitro reconstitution system for Tetrahymena telomerase and tested the effects of amino acid substitutions on activity. Substitution of the retroviral-like tyrosine in motif C did not change overall enzymatic activity but increased processivity. This increase in processivity correlated with an increased affinity for telomeric DNA primer. Substitution of an alanine did not increase processivity, while substitution of a phenylalanine had an intermediate effect. The data suggest that this amino acid is involved in interactions with the primer in telomerase as in other RTs, and show that mutating an amino acid to that conserved in retroviral RTs makes telomerase more closely resemble these other RTs. PMID- 10807926 TI - Interaction of the EEA1 FYVE finger with phosphatidylinositol 3-phosphate and early endosomes. Role of conserved residues. AB - FYVE zinc finger domains, which are conserved in multiple proteins from yeast to man, interact specifically with the membrane lipid phosphatidylinositol 3 phosphate (PtdIns(3)P). Here we have investigated the structural requirements for the interaction of the FYVE finger of the early endosome antigen EEA1 with PtdIns(3)P and early endosomes. The binding of the FYVE finger to PtdIns(3)P is Zn(2+)-dependent, and Zn(2+) could not be replaced by any other bivalent cations tested. By surface plasmon resonance, the wild-type FYVE finger was found to bind to PtdIns(3)P with an apparent K(D) of about 50 nm and a 1:1 stoichiometry. Mutagenesis of cysteines involved in Zn(2+) coordination, basic residues thought to be directly involved in ligand binding and other conserved residues, resulted in a 6- to >100-fold decreased affinity for PtdIns(3)P. A mutation in the putative PtdIns(3)P-binding pocket, R1375A, may prove particularly informative, because it led to a strongly decreased affinity for PtdIns(3)P without affecting the FYVE three-dimensional structure, as measured by fluorescence spectroscopy. Whereas the C terminus of EEA1 localizes to early endosomes when expressed in mammalian cells, all the FYVE mutants with reduced affinity for PtdIns(3)P were found to be largely cytosolic. Furthermore, whereas expression of the wild-type EEA1 C terminus interferes with early endosome morphology, the point mutants were without detectable effect. These results support recently proposed models for the ligand binding of the FYVE domain and indicate that PtdIns(3)P binding is crucial for the localization and function of EEA1. PMID- 10807927 TI - Direct and GTP-dependent interaction of ADP-ribosylation factor 1 with clathrin adaptor protein AP-1 on immature secretory granules. AB - ADP-ribosylation factor 1 (ARF1) mediates clathrin coat formation on PC12 immature secretory granules (ISGs). We have used two approaches to investigate whether ARF1 interacts directly with the clathrin adaptor protein, AP-1. Using an in vitro recruitment assay and co-immunoprecipitation, we could isolate an AP 1.ARF1 complex. Then we used a site-directed photocross-linking approach to determine the components that act downstream of ARF1 in clathrin coat formation on ISGs. Myristoylated ARF1, with a photolabile phenylalanine analogue incorporated into its putative effector domain (switch 1), showed a specific, GTP dependent interaction with both the gamma- and beta-adaptin subunits of AP-1 on ISGs. These experiments provide evidence for a direct interaction of ARF1 with AP 1. On mature secretory granules myristoylated ARF1 does not bind, and hence clathrin coat formation cannot be initiated, supporting the hypothesis that molecules involved in coat recruitment are removed during ISG maturation. PMID- 10807928 TI - Evidence for post-translational membrane insertion of the integral membrane protein bacterioopsin expressed in the heterologous halophilic archaeon Haloferax volcanii. AB - The gene coding for the integral membrane protein bacterioopsin (Bop), that is composed of seven transmembrane helices, was expressed in the halophilic archaeon Haloferax volcanii as a fusion protein with the halobacterial enzyme dihydrofolate reductase and with the cellulose binding domain of Clostridium thermocellum cellulosome. In each case, bacterioopsin was present both in the membrane and in the cytoplasmic fractions. Pulse-chase labeling experiments showed that the fusion protein in the cytoplasmic fraction is the precursor of the membrane-bound species. Bacterioopsin mutants that lack the seventh helix (BopDelta7) were found to accumulate only in the cytoplasmic fraction, whereas bacterioopsin mutants that lack either helices four and five (BopDelta4-5), or helices one and two (BopDelta1-2), were found in the cytoplasmic as well as in the membrane fractions. The seventh helix, when expressed alone, could target in trans the insertion of a separately expressed bacterioopsin mutant protein that has only the first six helices. These results support a model in which bacterioopsin is produced in H. volcanii as a soluble protein and in which its insertion into the membrane occurs post-translationally. According to this model, membrane insertion is directed by the seventh helix. PMID- 10807929 TI - A revised model for the structure and function of the lactose permease. Evidence that a face on transmembrane segment 2 is important for conformational changes. AB - The lactose permease is an integral membrane protein that cotransports H(+) and lactose into the bacterial cytoplasm. Previous work has shown that bulky substitutions at glycine 64, which is found on the cytoplasmic edge of transmembrane segment 2 (TMS-2), cause a substantial decrease in the maximal velocity of lactose uptake without significantly affecting the K(m) values (Jessen-Marshall, A. E., Parker, N. J., and Brooker, R. J. (1997) J. Bacteriol. 179, 2616-2622). In the current study, mutagenesis was conducted along the face of TMS-2 that contains glycine-64. Single amino acid substitutions that substantially changed side-chain volume at codons 52, 57, 59, 63, and 66 had little or no effect on transport activity, whereas substitutions at codons 49, 53, 56, and 60 were markedly defective and/or had lower levels of expression. According to helical wheel plots, Phe-49, Ser-53, Ser-56, Gln-60, and Gly-64 form a continuous stripe along one face of TMS-2. Several of the TMS-2 mutants (S56Y, S56L, S56Q, Q60A, and Q60V) were used as parental strains to isolate mutants that restore transport activity. These mutations were either first-site mutations or second-site suppressors in TMS-1, TMS-2, TMS-7 or TMS-11. A kinetic analysis showed that the suppressors had a higher rate of lactose transport compared with the corresponding parental strains. Overall, the results of this study are consistent with the notion that a face on TMS-2, containing Phe-49, Ser-53, Ser 56, Gln-60, and Gly-64, plays a critical role in conformational changes associated with lactose transport. We hypothesize that TMS-2 slides across TMS-7 and TMS-11 when the lactose permease interconverts between the C1 and C2 conformations. This idea is discussed within the context of a revised model for the structure of the lactose permease. PMID- 10807930 TI - 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced c-Jun N-terminal kinase (JNK) phosphatase renders immortalized or transformed epithelial cells refractory to TPA-inducible JNK activity. AB - c-Jun N-terminal kinase (JNK) regulates gene expression in response to various extracellular stimuli. JNK can be activated by the tumor promoting agent, 12-O tetradecanoylphorbol-13-acetate (TPA) in normal human oral keratinocytes but not in human keratinocytes that have been immortalized (HOK-16B and HaCaT) or transformed (HOK-16B-Bap-T) nor in a cervical carcinoma cell line (HeLa). The refractory JNK activation response to TPA is not due a defect in the JNK pathway, because JNK can be activated by other stimuli, e.g. UV irradiation and an alkylating agent N-methyl-N'-nitrosoguanidine in these immortalized or transformed cells. More importantly, the refractory JNK and JNKK activation response to TPA can be restored by treatment of the cells with a combination of TPA and a protein-tyrosine phosphatase inhibitor, sodium orthovanadate. Furthermore, pretreatment of cells with TPA partially inhibited UV- or N-methyl N'-nitrosoguanidine-induced JNK activity. These results suggest that a TPA inducible, orthovanadate-sensitive protein-tyrosine phosphatase may specifically down-regulate JNK signaling pathway in these immortalized/transformed epithelial cells. In contrast, ERK and p38/Mpk2 are not regulated by this TPA-induced phosphatase. This putative protein-tyrosine phosphatase appears to be JNK pathway specific. PMID- 10807931 TI - The phosphatase activity is the target for Mg2+ regulation of the sensor protein PhoQ in Salmonella. AB - The PhoP/PhoQ two-component system controls the expression of essential virulence traits in the pathogenic bacterium Salmonella enterica serovar Typhimurium. Environmental deprivation of Mg(2+) activates the PhoP/PhoQ signal transduction cascade, which results in an increased expression of genes necessary for survival inside the host. It was previously demonstrated that the interaction of Mg(2+) with the periplasmic domain of PhoQ promotes a conformational change in the sensor protein that leads to the down-regulation of PhoP-activated genes. We have now examined the regulatory effect of Mg(2+) on the putative activities of the membrane-bound PhoQ. We demonstrated that Mg(2+) promotes a phospho-PhoP phosphatase activity in the sensor protein. This activity depends on the intactness of the conserved His-277, suggesting that the phosphatase active site overlaps the H box. The integrity of the N-terminal domain of PhoQ was essential for the induction of the phosphatase activity, because Mg(2+) did not stimulate the release of inorganic phosphate from phospho-PhoP in a fusion protein that lacks this sensing domain. These findings reveal that the sensor PhoQ harbors a phospho-PhoP phosphatase activity, and that this phosphatase activity is the target of the extracellular Mg(2+)-triggered regulation of the PhoP/PhoQ system. PMID- 10807932 TI - Regulation of interleukin-1beta -induced platelet-derived growth factor receptor alpha expression in rat pulmonary myofibroblasts by p38 mitogen-activated protein kinase. AB - The potential role of p38 mitogen-activated protein (MAP) kinase in platelet derived growth factor receptor-alpha (PDGF-Ralpha) gene expression was investigated using cultured rat pulmonary myofibroblasts. p38 MAP kinase was constitutively expressed in myofibroblasts and activated by interleukin (IL) 1beta. A pyridinylimidazole compound, SB203580, completely inhibited the ability of p38 MAP kinase activity to phosphorylate PHAS-1 substrate. SB203580 inhibited IL-1beta-induced up-regulation of PDGF-Ralpha mRNA and protein in a concentration dependent manner. Other kinase inhibitors, including the mitogen-activated protein kinase/extracellular signal-regulated kinase inhibitor PD98059, did not block up-regulation of PDGF-Ralpha. The IL-1beta-induced increase in the number of (125)I-PDGF-AA-binding sites at the cell surface was reduced >70% by pretreatment with SB203580. Accordingly, an enhancement of PDGF-AA-stimulated DNA synthesis following IL-1beta pretreatment was blocked >70% by SB203580. SB203580 did not affect IL-1beta-induced ERK activation, yet enhanced IL-1beta-induced JNK activation approximately 2-fold. Treatment of cells with SB203580 after inhibition of transcription by actinomycin D decreased the half-life of IL-1beta induced PDGF-Ralpha mRNA from >4 to approximately 1.5 h. Moreover, pretreatment of cells with cycloheximide blocked induction of PDGF-Ralpha mRNA by IL-1beta, suggesting that de novo protein synthesis was required for PDGF-Ralpha mRNA stabilization. These data indicate that p38 MAP kinase regulates PDGF-Ralpha expression at the translational level by signaling the synthesis of an mRNA stabilizing protein. PMID- 10807933 TI - Activation of NF-kappa B by XIAP, the X chromosome-linked inhibitor of apoptosis, in endothelial cells involves TAK1. AB - Exposure of endothelial and many other cell types to tumor necrosis factor alpha generates both apoptotic and anti-apoptotic signals. The anti-apoptotic pathway leads to activation of the transcription factor NF-kappaB that regulates the expression of genes such as A20 or members of the IAP gene family that protect cells from tumor necrosis factor alpha-mediated apoptosis. In turn, some anti apoptotic genes have been shown to modulate NF-kappaB activity. Here we demonstrate that XIAP, a NF-kappaB-dependent member of the IAP gene family, is a strong stimulator of NF-kappaB. Expression of XIAP leads to increased nuclear translocation of the p65 subunit of NF-kappaB via a novel signaling pathway that involves the mitogen-activated protein kinase kinase kinase TAK1. We show that TAK1 physically interacts with NIK and with IKK2, and both XIAP or active TAK1 can stimulate IKK2 kinase activity. Thus, XIAP may be part of a system of regulatory loops that balance a cell's response to environmental stimuli. PMID- 10807934 TI - The regulator of G protein signaling RGS4 selectively enhances alpha 2A adreoreceptor stimulation of the GTPase activity of Go1alpha and Gi2alpha. AB - Agonist-stimulated high affinity GTPase activity of fusion proteins between the alpha(2A)-adrenoreceptor and the alpha subunits of forms of the G proteins G(i1), G(i2), G(i3), and G(o1), modified to render them insensitive to the action of pertussis toxin, was measured following transient expression in COS-7 cells. Addition of a recombinant regulator of G protein signaling protein, RGS4, did not significantly affect basal GTPase activity nor agonist stimulation of the fusion proteins containing Galpha(i1) and Galpha(i3) but markedly enhanced agonist stimulation of the proteins containing Galpha(i2) and Galpha(o1.) The effect of RGS4 on the alpha(2A)-adrenoreceptor-Galpha(o1) fusion protein was concentration dependent with EC(50) of 30 +/- 3 nm and the potency of the receptor agonist UK14304 was reduced 3-fold by 100 nm RGS4. Equivalent reconstitution with Asn(88) Ser RGS4 failed to enhance agonist function on the alpha(2A)-adrenoreceptor Galpha(o1) or alpha(2A)-adrenoreceptor-Galpha(i2) fusion proteins. Enzyme kinetic analysis of the GTPase activity of the alpha(2A)-adrenoreceptor-Galpha(o1) and alpha(2A)-adrenoreceptor-Galpha(i2) fusion proteins demonstrated that RGS4 both substantially increased GTPase V(max) and significantly increased K(m) of the fusion proteins for GTP. The increase in K(m) for GTP was dependent upon RGS4 amount and is consistent with previously proposed mechanisms of RGS function. Agonist-stimulated GTPase turnover number in the presence of 100 nm RGS4 was substantially higher for alpha(2A)-adrenoreceptor-Galpha(o1) than for alpha(2A) adrenoreceptor-Galpha(i2). These studies demonstrate that although RGS4 has been described as a generic stimulator of the GTPase activity of G(i)-family G proteins, selectivity of this interaction and quantitative variation in its function can be monitored in the presence of receptor activation of the G proteins. PMID- 10807935 TI - Integrins minireview series. PMID- 10807936 TI - Involvement of the proximal C terminus of the AMPA receptor subunit GluR1 in dendritic sorting. AB - Studies on dendritic sorting of transmembrane proteins in hippocampal neurons in culture have shown that these cells use similar mechanisms as epithelial cells to sort transmembrane proteins to the basolateral membrane domain. However, information is still scarce with regard to which amino acidic sequences are required for dendritic sorting in neurons. The glutamate receptor 1 (GluR1) subunit of the AMPA receptor is present on the dendritic compartment of hippocampal neurons in culture. To identify the GluR1 sorting signal responsible for dendritic targeting, we have expressed the wild-type GluR1, a deletion mutant in the C-terminal cytoplasmic tail, and chimeric GluR1 proteins in hippocampal neurons using a calcium phosphate transfection method. The recombinant full length GluR1 is polarized to the dendritic domain. Truncated GluR1 with a deletion of the C-terminal cytoplasmic tail is still delivered to the somatodendritic domain. However a chimeric protein made of the luminal and transmembrane domain of the influenza virus hemagglutinin (HA) fused to the GluR1 C-terminal cytoplasmic tail (HaemR1) is detected in the somatodendritic domain. This finding indicates that the GluR1 C-terminal cytoplasmic tail contains a dendritic sorting signal, which redirects the axonal or axonal-dendritic protein HA to the dendritic compartment exclusively. Deletion analysis of HaemR1 shows that the proximal segment of the GluR1 C-terminal cytoplasmic tail contains a novel dendritic sorting signal. PMID- 10807937 TI - Accumulation of zinc in degenerating hippocampal neurons of ZnT3-null mice after seizures: evidence against synaptic vesicle origin. AB - In several brain injury models, zinc accumulates in degenerating neuronal somata. Suggesting that such zinc accumulation may play a causal role in neurodegeneration, zinc chelation attenuates neuronal death. Because histochemically reactive zinc is present in and released from synaptic vesicles of glutamatergic neurons in the forebrain, it was proposed that zinc translocation from presynaptic terminals to postsynaptic neurons may be the mechanism of toxic zinc accumulation. To test this hypothesis, kainate seizure induced neuronal death was examined in zinc transporter 3 gene (ZnT3)-null mice, a strain that completely lacks histochemically reactive zinc in synaptic vesicles. Intraperitoneal injection of kainate induced seizures to a similar degree in wild type and ZnT3-null mice. Staining of hippocampal sections with a zinc-specific fluorescent dye, N-(6-methoxy-8-quinolyl)-p carboxybenzoylsulfonamide, revealed that zinc accumulated in degenerating CA1 and CA3 neurons in both groups, indicating that zinc originated from sources other than synaptic vesicles. Injection of CaEDTA into the cerebral ventricle almost completely blocked zinc accumulation in ZnT3-null mice, suggesting that increases in extracellular zinc concentrations may be a critical event for zinc accumulation. Arguing against the possibility that zinc accumulation results from nonspecific breakdown of zinc-containing proteins, injection of kainate into the cerebellum did not induce zinc accumulation in degenerating granule neurons. Taken together, these results support the existing idea that zinc is released into extracellular space and then enters neurons to exert a cytotoxic effect. However, the origin of zinc is not likely to be synaptic vesicles, because zinc accumulation robustly occurs in ZnT3-null mice lacking synaptic vesicle zinc. PMID- 10807939 TI - Dietary soy-derived isoflavone phytoestrogens. Could they have a role in coronary heart disease prevention? AB - Soy protein-containing foods are a rich source of isoflavone phytoestrogens, such as genistein and daidzein. There is great interest in these substances, as lower rates of chronic diseases, including coronary heart disease, have been associated with high dietary intake of soy-containing foods. Soy phytoestrogens bind weakly to estrogen receptors, and some bind more strongly to estrogen receptor-beta compared with estrogen receptor-alpha. A meta-analysis has indicated that isoflavone phytoestrogens lowered plasma cholesterol concentrations in subjects with initially elevated levels, but had little effect in subjects with normal cholesterol concentrations. These substances reportedly may also have beneficial effects on arterial endothelial function. In addition to these potentially antiatherogenic effects, many laboratories are investigating other possible mechanisms, including antioxidative and antiproliferative properties of these substances. We have shown that dietary supplementation with soy-derived isoflavones reduced the in vitro oxidation susceptibility of low-density lipoprotein (LDL). To further explore this phenomenon, we incorporated genistein and daidzein into LDL molecules in vitro with the aid of an artificial transfer system. However, it was necessary to convert the isoflavone molecules to fat soluble derivatives, fatty acid esters (analogous to esterified endogenous estrogens, which are known to occur in vivo), to achieve significant incorporation. The LDLs containing esterified isoflavones were shown to be less susceptible to oxidation in vitro than native LDL. We also employed U937 cell cultures for investigating the effects of isoflavone-containing LDLs on cell proliferation. Some of these LDLs exhibited antiproliferative effects in cultured U937 cells. In summary, lipophilic phytoestrogen derivatives could be incorporated into LDLs, increasing their oxidation resistance and antiproliferative efficacy ex vivo, both of which are, in theory, antiatherogenic effects. Further studies are needed to assess to what extent analogous effects could be produced in vivo and whether such substances have a role in hormone replacement and coronary heart disease prevention in postmenopausal women. PMID- 10807940 TI - Stereoselective sulfoxidation of sulindac sulfide by flavin-containing monooxygenases. Comparison of human liver and kidney microsomes and mammalian enzymes. AB - The stereoselective sulfoxidation of the pharmacologically active metabolite of sulindac, sulindac sulfide, was characterized in human liver, kidney, and cDNA expressed enzymes. Kinetic parameter estimates (pH = 7.4) for sulindac sulfoxide formation in human liver microsomes (N = 4) for R- and S-sulindac sulfoxide were V(max) = 1.5 +/- 0.50 nmol/min/mg, K(m) = 15 +/- 5.1 microM; and V(max) = 1.1 +/- 0.36 nmol/min/mg, K(m) = 16 +/- 6.1 microM, respectively. Kidney microsomes (N = 3) produced parameter estimates (pH = 7.4) of V(max) = 0.9 +/- 0.29 nmol/min/mg, K(m) = 15 +/- 2.9 microM; V(max) = 0.5 +/- 0.21 nmol/min/mg, K(m) = 22 +/- 1.9 microM for R- and S-sulindac sulfoxide, respectively. In human liver and flavin containing monooxygenase 3 (FMO3) the V(max) for R-sulindac sulfoxide increased 60-70% at pH = 8.5, but for S-sulindac sulfoxide was unchanged. In fourteen liver microsomal preparations, significant correlations occurred between R-sulindac sulfoxide formation and either immunoquantified FMO or nicotine N-oxidation (r = 0.88 and 0.83; P < 0.01). The R- and S-sulindac sulfoxide formation rate also correlated significantly (r = 0.85 and 0.75; P < 0.01) with immunoquantified FMO in thirteen kidney microsomal samples. Mild heat deactivation of microsomes reduced activity by 30-60%, and a loss in stereoselectivity was observed. Methimazole was a potent and nonstereoselective inhibitor of sulfoxidation in liver and kidney microsomes. n-Octylamine and membrane solubilization with lubrol were potent and selective inhibitors of S-sulindac sulfoxide formation. cDNA expressed CYPs failed to appreciably sulfoxidate sulindac sulfide, and CYP inhibitors were ineffective in suppressing catalytic activity. Purified mini-pig liver FMO1, rabbit lung FMO2, and human cDNA-expressed FMO3 efficiently oxidized sulindac sulfide with a high degree of stereoselectivity towards the R-isomer, but FMO5 lacked catalytic activity. The biotransformation of the sulfide to the sulfoxide is catalyzed predominately by FMOs and may prove to be useful in characterizing FMO activity. PMID- 10807941 TI - Status of glutathione and other thiols and disulfides in human plasma. AB - While plasma thiols, including homocysteine (HCys), glutathione (GSH), and cysteine (Cys), are being investigated as potential indicators of disease risk and health status, low levels, poor stability, and the lack of comprehensive methodologies have hampered their accurate assessment. Using our previously described HPLC with electrochemical detection method, our goal was to assess levels, stability, and distribution of biologically relevant thiols and disulfides in human plasma. In fresh plasma, processed immediately after collection, low levels of Cys, cystine, Cys-Gly, and the mixed disulfide Cys-GSH (CSSG) were consistently observed, whereas the levels of GSH and Cys-Gly disulfide were often below the limits of detection. These profiles were a consequence of poor thiol stability, as thiol standards added to human plasma were lost rapidly due to autoxidation or formation of mixed disulfides. A 75% loss of added GSH observed after 30 min was accounted for completely by the formation of GSH disulfide (24%) and CSSG (74%). Similar changes were found with other thiols when added to plasma. Thiols lost to oxidation were recovered quantitatively by reducing samples with potassium borohydride (KBH(4)) prior to analysis. In a study of 106 healthy adults, mean total thiol levels in plasma were: Cys (201 microM) > Cys-Gly (101 microM) > HCys (7 microM) > gamma-Glu-Cys (5 microM) > GSH (4 microM). All together, these results account for the poor stability of thiols in plasma and provide a method for their comprehensive and accurate determination. PMID- 10807942 TI - Induction of erythroid differentiation of human K562 cells by cisplatin analogs. AB - Human leukemic K562 cells can be induced in vitro to erythroid differentiation by a variety of chemical compounds, including hemin, butyric acid, 5-azacytidine, and cytosine arabinoside. Differentiation of K562 cells is associated with an increase in the expression of embryo-fetal globin genes, such as the zeta-, epsilon-, and gamma-globin genes. Therefore, the K562 cell line has been proposed as a very useful in vitro model system for determining the therapeutic potential of new differentiating compounds as well as for studying the molecular mechanism(s) regulating changes in the expression of embryonic and fetal human globin genes. Inducers of erythroid differentiation that stimulate gamma-globin synthesis could be considered for possible use in the experimental therapy of hematological diseases associated with a failure in the expression of adult beta globin genes. In this paper, we analyzed the effects of a series of cisplatin analogs on both cell growth and differentiation of K562 cells. Among seven cisplatin analogs studied, three were found to be potent inducers of erythroid differentiation. Erythroid differentiation was associated with an increase in the accumulation of (a) hemoglobins Gower 1 and Portland and (b) gamma-globin mRNA. PMID- 10807943 TI - Efficient utilization of the reduced folate carrier in CCRF-CEM human leukemic lymphoblasts by the potent antifolate N(alpha)-(4-amino-4-deoxypteroyl)-N(delta) hemiphthaloyl-L- ornithine (PT523) and its B-ring analogues. AB - The potent nonpolyglutamatable dihydrofolate reductase inhibitor N(alpha)-(4 amino-4-deoxypteroyl)-N(delta)-hemiphthaloyl-L-o rnithine (PT523) and six of its B-ring (5-deaza, 8-deaza, and 5,8-dideaza) analogues were compared in terms of their ability to: (a) inhibit the growth of CCRF-CEM human leukemic lymphoblasts, and (b) utilize the reduced folate carrier (RFC) in these cells as measured in a competition assay of [(3)H]methotrexate ([(3)H]MTX) influx. The IC(50) values of the hemiphthaloylornithine derivatives against CCRF-CEM cells after 72 hr of drug exposure varied from 0.64 to 1.3 nM as compared with 14 nM for MTX and 4.4 nM for aminopterin (AMT). The K(i) values of these compounds in the [(3)H]MTX influx assay were in the 0.3 to 0.7 microM range as compared with a K(i) of 5.4 microM for AMT and a K(t) of 7.1 microM for MTX. As a group, the affinities of these compounds for the RFC were approximately 10-fold greater than those of their respective glutamate analogues. These results indicate that, in addition to their previously reported tight binding to dihydrofolate reductase, a property contributing to the high potency of PT523 and its B-ring analogs as inhibitors of tumor cell growth is their strong affinity for the RFC. PMID- 10807944 TI - Ascorbate-dependent protection of human erythrocytes against oxidant stress generated by extracellular diazobenzene sulfonate. AB - Diazobenzene sulfonic acid (DABS) has been used to label thiols and amino groups on cell-surface proteins. However, we found that in addition to inhibiting an ascorbate-dependent trans-plasma membrane oxidoreductase in human erythrocytes, it also depleted alpha-tocopherol severely in the cell membrane. When erythrocytes were loaded with ascorbate, DABS-dependent loss of alpha-tocopherol was decreased, despite little change in intracellular ascorbate content. Sparing of alpha-tocopherol also was seen in erythrocyte ghosts resealed to contain ascorbate, although this was accompanied by loss of intravesicular ascorbate, probably due to the inability of ghosts to recycle ascorbate. A transmembrane transfer of electrons from ascorbate was confirmed by electron paramagnetic resonance spectroscopy, in which extracellular DABS was found to generate the ascorbate free radical within cells. When the membrane content of alpha tocopherol was decreased to 20% of the initial value by DABS treatment, lipid peroxidation ensued, manifest by generation of F(2)-isoprostanes in the cell membranes. Intracellular ascorbate also strongly protected against F(2) isoprostane formation. These results show that DABS causes an oxidant stress at the membrane surface that is transmitted within the cell, in part by an alpha tocopherol-dependent mechanism, and that ascorbate recycling of alpha-tocopherol can protect against loss of alpha-tocopherol and the ensuing lipid peroxidation. PMID- 10807945 TI - Kinetics and mechanism of the reaction of aminoguanidine with the alpha oxoaldehydes glyoxal, methylglyoxal, and 3-deoxyglucosone under physiological conditions. AB - Aminoguanidine (AG), a prototype agent for the preventive therapy of diabetic complications, reacts with the physiological alpha-oxoaldehydes glyoxal, methylglyoxal, and 3-deoxyglucosone (3-DG) to form 3-amino-1,2,4-triazine derivatives (T) and prevent glycation by these agents in vitro and in vivo. The reaction kinetics of these alpha-oxoaldehydes with AG under physiological conditions pH 7.4 and 37 degrees was investigated. The rate of reaction of AG with glyoxal was first order with respect to both reactants; the rate constant k(AG,G) was 0.892 +/- 0.037 M(-1) sec(-1). The kinetics of the reaction of AG with 3-DG were more complex: the rate equation was d[T](o)/dt (initial rate of T formation) = [3-DG](k(AG,3-DG)[AG] + k(3-DG)), where k(AG,3-DG) = (3. 23 +/- 0.25) x 10(-3) M(-1) sec(-1) and k(3-DG) = (1.73 +/- 0.08) x 10(-5) sec(-1). The kinetics of the reaction of AG with methylglyoxal were consistent with the reaction of both unhydrated (MG) and monohydrate (MG-H(2)O) forms. The rate equation was d[T](o)/dt = ?k(1)k(AG,MG)/(k(-1) + k(AG,MG)[AG]) + k(AG, MG H(2)O)?[MG-H(2)O][AG], where the rate constant for the reaction of AG with MG, k(AG,MG), was 178 +/- 15 M(-1) sec(-1) and for the reaction of AG with MG-H(2)O, k(AG,MG-H(2)O), was 0.102 +/- 0.001 M(-1) sec(-1); k(1) and k(-1) are the forward and reverse rate constants for methylglyoxal dehydration MG-H(2)O right harpoon over left harpoon MG. The kinetics of these reactions were not influenced by ionic strength, but the reaction of AG with glyoxal and with methylglyoxal under MG-H(2)O dehydration rate-limited conditions increased with increasing phosphate buffer concentration. Kinetic modelling indicated that the rapid reaction of AG with the MG perturbed the MG/MG-H(2)O equilibrium, and the ratio of the isomeric triazine products varied with initial reactant concentration. AG is kinetically competent to scavenge the alpha-oxoaldehydes studied and decrease related advanced glycated endproduct (AGE) formation in vivo. This effect is limited, however, by the rapid renal elimination of AG. Decreased AGE formation is implicated in the prevention of microvascular complications of diabetes by AG. PMID- 10807946 TI - Down-regulation by troglitazone of hepatic tumor necrosis factor-alpha and interleukin-6 mRNA expression in a murine model of non-insulin-dependent diabetes. AB - Troglitazone, a novel thiazolidinedione drug used to treat non-insulin-dependent diabetes mellitus, is a selective ligand for the peroxisome proliferator activated receptor-gamma (PPARgamma). Recent results indicate that PPARgamma activation by thiazolidinediones regulates adipose tissue- and monocyte/peritoneal macrophage-derived cytokine expression in vitro. We evaluated whether troglitazone may also negatively regulate cytokine expression in the liver, which harbors the majority of the body's resident macrophages but which only weakly expresses PPARgamma. Lean C57BL6 mice and genetically obese KKA(y) mice were chronically treated with troglitazone (100 mg/kg/day for 2 weeks). At the end of treatment, hepatic expression of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 mRNA was quantitatively determined by kinetic polymerase chain reaction both under basal conditions and after stimulation with lipopolysaccharide (LPS). Both untreated lean and obese mice exhibited low levels of baseline TNF-alpha and IL-6 mRNA expression and responded with a dramatic increase in hepatic cytokine transcripts and TNF-alpha protein expression following a challenge with LPS. Similar to the effects on white adipose tissue, troglitazone not only down-regulated the baseline levels of hepatic TNF-alpha and IL-6, but also greatly attenuated the inducing effects of LPS. The extent of this inhibitory effect of troglitazone was higher in obese KKA(y) mice than in lean mice and was also reflected by markedly down-regulated hepatic TNF-alpha protein expression. These data demonstrate that chronic administration of troglitazone is associated with a greatly attenuated responsiveness towards inducers of hepatic TNF-alpha and IL-6 production. The possible biological consequences of these effects, however, have not yet been assessed. PMID- 10807947 TI - Bcl-2 phosphorylation in a human breast carcinoma xenograft: a common event in response to effective DNA-damaging drugs. AB - A variety of cytotoxic agents effective as antitumor drugs are known to kill tumor cells through induction of apoptosis as the most relevant modality of cell death. A specific role for the protein Bcl-2 in the cell death pathway induced by antimicrotubule agents has been proposed, because Bcl-2 phosphorylation occurs in response to microtubule damage. In this study, we compared efficacy, apoptosis, and Bcl-2 phosphorylation in the Bcl-2-overexpressing MX-1 human breast carcinoma xenograft after treatment with cytotoxic agents characterized by different mechanisms of action. We demonstrated that, in addition to antimicrotubule agents, effective DNA-damaging agents were also able to induce Bcl-2 phosphorylation irrespective of the type of genotoxic lesion. A comparison of effects of drugs belonging to the same class but endowed with a different antitumor activity (i.e. cisplatin versus a novel multinuclear platinum complex and doxorubicin versus a disaccharide analogue) showed a correlation between drug efficacy, apoptotic response, and Bcl-2 phosphorylation. In conclusion, overexpression of Bcl-2 did not counteract the apoptotic effects of a number of cytotoxic agents and could not be regarded as a mechanism of cellular resistance. Since Bcl-2 phosphorylation is a common event in response to different types of cytotoxic damage and is not only related to microtubule dysfunction, we suggest that many cell death pathways converge on Bcl-2 and protein phosphorylation is a step of the signaling cascade activated by diverse stimuli and likely related to the onset of drug-induced apoptosis. PMID- 10807948 TI - Antisense inhibition of P-glycoprotein expression using peptide-oligonucleotide conjugates. AB - Antisense oligonucleotides are potentially a powerful tool for the therapeutic manipulation of genes associated with cancer. However, pharmacological applications of oligonucleotides have been hindered by the inability to effectively deliver these compounds to their sites of action within cells. In this study, we have prepared peptide-oligonucleotide conjugates with the intent of improving intracellular delivery. The phosphorothioate oligonucleotide component of the conjugates was complementary to a site flanking the AUG of the message for P-glycoprotein, a membrane ATPase associated with multidrug resistance in tumor cells. Two types of peptide-antisense oligonucleotide conjugates, but not mismatched control conjugates, provided substantial inhibition of cell surface expression of P-glycoprotein. Surprisingly, the peptide-oligonucleotide conjugates were more potent in the presence of serum than when used under serum-free conditions; this is in striking contrast to most other approaches for intracellular delivery of nucleic acids. Effective inhibition of P glycoprotein expression was attained with submicromolar concentrations of antisense conjugates under serum-replete conditions. The combination of relatively modest molecular size and good efficacy in the presence of serum proteins suggests that peptide-antisense oligonucleotide conjugates may have significant promise for in vivo therapeutic applications. PMID- 10807949 TI - Additive inhibitory effect of calcipotriol and anthralin on ribonuclease P activity. AB - The effects of two antipsoriatic compounds, calcipotriol and anthralin, separately or in combination on ribonuclease P (RNase P), were investigated using a cell-free system from the slime mold Dictyostelium discoideum. RNase P is an ubiquitous and essential enzyme which endonucleolytically cleaves all tRNA precursors to produce the mature 5' end. The substrate for RNase P assays was an in vitro (32)P-labeled transcript of the Schizosaccharomyces pombe tRNA(Ser) gene supS1. Enzyme assays were carried out at 37 degrees in 20 microL 50 mM Tris-HCL 7.6 buffer, containing 10 mM NH(4)Cl, 5 mM MgCl(2), and 10% isopropanol. Calcipotriol or anthralin alone exerted a dose-dependent inhibitory effect on RNase P activity, with the former being more active than the latter in this respect. Simultaneous exposure of the enzyme to both drugs resulted in an enhancement of RNase P inhibition, which was additive. Considering the lack of structural similarities between the substrate (precursor tRNA) of RNase P and the tested drugs, it seems reasonable to suggest that their effects may be due to binding to allosteric inhibition sites of the enzyme. Although our in vitro findings cannot be directly extrapolated to the in vivo human condition, they do suggest that the inhibitory effects of calcipotriol and anthralin on tRNA biogenesis may be implicated in the mechanisms of their antipsoriatic action. Moreover, the additive inhibitory effect of these compounds on RNase P activity provides an experimental basis for their possible combined therapeutic application in the management of psoriasis. PMID- 10807950 TI - Inactivation of creatine kinase by Adriamycin during interaction with horseradish peroxidase. AB - Oxidative damage of creatine kinase (CK) induced by Adriamycin((R)) (ADM) with peroxidase was investigated using horseradish peroxidase (HRP). ADM oxidatively inactivated CK during its interaction with HRP in the presence of H(2)O(2) (HRP H(2)O(2)). The red color of ADM was lost during oxidation by HRP-H(2)O(2). Adding catalase stopped the color change of ADM induced by HRP-H(2)O(2), indicating that ADM was oxidized by HRP complex I or II. CK was inactivated readily, even when it was added to the reaction mixture containing colorless ADM. Some sulfhydryl groups of CK, which have an important role in its enzyme activity, were very sensitive to ADM activated by HRP-H(2)O(2), suggesting that inactivation of CK is due to oxidation of SH groups at the active center. Presumably, oxidative ADM quinone is involved dominantly in the inactivation of CK. Among the anthracycline drugs tested in this study, only ADM and epirubicin caused inactivation of CK and alcohol dehydrogenase and loss of the red color during oxidation by HRP-H(2)O(2). PMID- 10807951 TI - Topoisomerase II cleavable complex formation within DNA loop domains. AB - The distribution of VM-26 (Teniposide)-stabilized cleavable complexes within DNA loops bound to the nuclear matrix was determined to provide further insights into the mode of DNA synthesis inhibition by VM-26. Covalent binding of [(3)H]VM-26 was 9-fold greater per milligram of nuclear matrix protein compared with high salt-soluble nonmatrix protein of CEM cells. The ratio declined from 9-fold in CEM cells to 4-fold in drug-resistant VM-1/C2 cells, which have decreased nuclear matrix DNA topoisomerase IIalpha. VM-26 induced a concentration-dependent increase in the frequency of cleavable complex formation with actively replicating matrix DNA. At 25 microM VM-26, the frequency was 32 +/- 2 (SEM) complexes per 10(6) bp of replicating matrix DNA compared with 13 +/- 2 (SEM) complexes per 10(6) bp of nonreplicating DNA in the matrix fraction. VM-26 at concentrations as high as 25 microM stabilized less than 3 complexes per 10(6) bp in the various nonmatrix DNA domains, since the nonmatrix DNA comprises the DNA loop domains that are distal to the matrix-bound replication sites. A negligible frequency of cleavable complex formation was detected in both the matrix and nonmatrix DNA domains of drug-resistant VM-1/C2 cells. Compared with untreated control cells, VM-26 induced an accumulation of nascent DNA in the nuclear matrix fraction of CEM cells but decreased the amount of nascent DNA in the nonmatrix fraction. The extensive cleavable complex formation on matrix replicating DNA stalled most of the replication forks within 1 kb of the replication sites on the nuclear matrix. The results provide evidence that nascent DNA bound to the nuclear matrix is an important site of VM-26 cleavable complex formation, and that these complexes inhibit DNA synthesis by blocking the movement of nascent DNA away from replication sites on the nuclear matrix. PMID- 10807952 TI - Apoptosis induction by a dopaminergic neurotoxin, 1-methyl-4-phenylpyridinium ion (MPP(+)), and inhibition by epidermal growth factor in GH3 cells. AB - A dopaminergic neurotoxin, 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine (MPTP), can induce dopaminergic denervation and Parkinsonism in humans. The active metabolite of MPTP is the 1-methyl-4-phenylpyridinium ion (MPP(+)). Previously we reported that MPP(+) is incorporated via the dopamine transport system and causes delayed cell death in GH3 cells, a clonal strain from the rat anterior pituitary. In this study, we investigated whether MPP(+) induces apoptosis. GH3 cells cultured with MPP(+) exhibited DNA laddering and fragmentation in a time- and concentration-dependent manner. The effect of MPP(+) was inhibited in GH3 cells treated with a pan-caspase inhibitor (100 microM ZVAD-fmk), an antioxidant (25 mM N-acetyl-l-cysteine), or epidermal growth factor (EGF; 50 ng/mL). Because EGF stimulated tyrosine phosphorylation of the EGF receptor and tyrphostin AG1478 [4 (3-chloroanilino)-6,7-dimethoxyquinazoline; 5 microM, a specific inhibitor of EGF receptor kinase] abolished EGF inhibition, involvement of EGF receptor kinase is assumed. Protein kinase C-dependent processes and Bcl-2 protein expression were shown not to be involved in EGF inhibition. MPP(+) increased cytochrome c immunoreactivity in cytosolic fractions in GH3 cells. The addition of 200 microM MPP(+) to isolated mitochondrial fractions from GH3 cells stimulated the release of a 13-kDa protein that cross-reacted with anti-cytochrome c antibody. The release was inhibited in EGF-treated GH3 cells. Our findings demonstrated that (i) MPP(+) induces apoptosis of GH3 cells via cytochrome c release and caspase activation, and (ii) apoptosis by MPP(+) can be blocked by N-acetyl-l-cysteine or EGF treatment. PMID- 10807953 TI - High-level expression of human butyrylcholinesterase gene in Bombyx mori and biochemical-pharmacological characteristic study of its product. AB - The human butyrylcholinesterase (BChE, EC 3.1.1.8) gene was highly expressed in Bombyx mori using baculovirus vector, and the biochemical-pharmacological properties of its product were studied. BChE cDNA was cloned into transfer vector pBn96 and co-transfected with wild-type Bombyx mori nucleopolyhedrovirus (BmNPV) DNA into BmN cells. The recombinant virus with the highest enzyme activity was sorted out and purified. Once the BmN cells or silkworm larvae had been infected with the recombinant virus, recombinant human BChE (rhBChE) could be secreted into the culture medium or the hemolymph of the larvae at levels of 1.5 mg x L( 1) and 35 mg x L(-1), respectively. Western blot and enzymatic staining of the electrophoresis gel of non-denatured protein showed that rhBChE manifested similar antigenicity and enzyme activity to native human BChE (nhBChE). The production of rhBChE in the hemolymph was 23-fold higher than that in BmN cells and about 280-fold that in Chinese hamster overy cells (125 microg x L(-1)). This is the first report of human BChE expression in silkworm with the highest level of yield so far. rhBChE was highly similar to nhBChE in respect to substrate affinity, inhibitor sensitivity, and reactivity of the inhibited enzyme. It is suggested that rhBChE functions as well as nhBChE and has potential practical value. PMID- 10807954 TI - Characterization of the mouse nuclear orphan receptor TR2-11 gene promoter and its potential role in retinoic acid-induced P19 apoptosis. AB - The complete mouse orphan nuclear receptor TR2-11 gene structure and its 5' untranscribed region were characterized. This gene contains 14 exons, with the first exon encoding only the 5'-untranslated sequence. The regulatory region of this gene was characterized by using reporter assays that define the minimal promoter activity in a sequence 212 nucleotides upstream from the translation initiation site. Furthermore, it was concluded that splicing of intron 1 is required for efficient promoter activity. Reporters driven by this promoter were induced by retinoic acid (RA) in COS-1 cells supplied with exogenous retinoic acid receptor-alpha (RAR(alpha)) and retinoid receptor X-beta (RXR(beta)). Binding of RAR(alpha)/RXR(beta) to the minimal promoter region was demonstrated in gel retardation assays. In P19 cells, both the endogenous TR2-11 gene and the reporters driven by this promoter were induced by RA in a protein synthesis independent manner, and overexpression of TR2-11 protein resulted in cellular apoptosis in the absence of RA. The regulation of TR2-11 by RA and the implication of TR2 up-regulation in P19 cellular apoptosis are discussed. PMID- 10807955 TI - Lack of a role for inducible nitric oxide synthase in an experimental model of nephrotic syndrome. AB - Puromycin aminonucleoside (PAN) administration in rats produces an experimental model of nephrotic syndrome characterized by glomerular epithelial cell injury and proteinuria. The purpose of this study was to examine the role of nitric oxide (NO) in this model of minimal change glomerular disease. Aminoguanidine (AG) was used to inhibit inducible nitric oxide synthase (iNOS). Sprague-Dawley rats were divided into Control (N = 9), PAN (N = 14), AG (N = 2), and PAN + AG (N = 12) treatment groups. Control animals received saline (i.v. ), PAN animals received PAN (75 mg/kg, i.v.), and PAN + AG animals received PAN plus AG (50 mg/kg, i.p., twice daily). AG animals received a saline injection (i.v.) on day 0 in the place of PAN and then AG on the same schedule as the PAN + AG group. Animals were kept in metabolic cages, and urinary protein excretion and nitrite (NO(2)(-)) excretion were measured daily. PAN administration increased urinary NO(2)(-) excretion by day 2, and levels remained elevated through day 7. AG prevented this PAN-induced increase in urinary NO(2)(-) excretion. Plasma nitrate (NO(3)(-)) and NO(2)(-) (NOx) concentrations were also increased in the PAN and PAN + AG groups. iNOS protein expression was not detected in either the glomeruli or the cortex at day 7. Proteinuria developed in PAN animals on day 4 and increased steadily through day 7. PAN + AG animals showed a pattern similar to that of the PAN group. These results indicated that in contrast to models of proliferative glomerulonephritis, NO formation during PAN-induced nephrotic syndrome is increased but does not participate in the development of glomerular injury as measured by proteinuria. PMID- 10807956 TI - Subcellular localization of neuronal nitric oxide synthase in rat small intestine. AB - The subcellular localization of neuronal nitric oxide synthase (NOS I, EC 1.14.13.39) was investigated in the longitudinal muscle/myenteric plexus (LM/MP) preparation of rat small intestine. The presence of NOS I, inducible nitric oxide synthase (NOS II), and endothelial nitric oxide synthase (NOS III) was assessed after homogenization and low-speed centrifugation in a postnuclear supernatant by immunological detection after PAGE and Western blotting. Only NOS I was clearly present, whereas NOS II and NOS III were below detection limits. After high-speed centrifugation of the postnuclear supernatant, soluble and particulate fractions were obtained, and the presence of NOS I in these fractions was investigated by measurement of NOS I immunoreactivity and enzyme activity. We found that 90 +/- 1% of NOS I immunoreactivity and 97 +/- 1% of NOS enzyme activity were confined to the soluble fraction of the tissue. Further immunological analysis demonstrated that washing the particulate fraction revealed detectable amounts of NOS I only after concentration of the washing supernatant. Most particulate NOS I remained in the pellet and therefore represents cell organelle-associated enzyme. No NOS I immunoreactivity could be detected as a soluble protein within organelles of the cell. Particulate NOS I could in part be solubilized by Triton X-100 treatment, and the detection of Triton X-100-soluble NOS I was dependent on the antibody used. In conclusion, our results indicate that NOS I in the LM/MP preparation of rat small intestine is mainly soluble and that the particulate NOS I is partly an intrinsic membrane protein and can partly be solubilized by detergent treatment. PMID- 10807957 TI - On defining Sydenham's chorea: where do we draw the line? AB - Sydenham's chorea (SC) is a major manifestation of rheumatic fever characterized by an array of neuropsychiatric symptoms that vary in severity, timing, and character. Some of the same symptoms are seen in Tourette's syndrome and childhood-onset obsessive-compulsive disorder. Genetic vulnerability appears to play a role in all three conditions. The term PANDAS (pediatric autoimmune neuropsychiatric disorder associated with streptococcus) has been introduced to describe a putative subset of obsessive-compulsive disorder and Tourette's syndrome that bears some resemblance to Sydenham's chorea. This article discusses whether PANDAS should be subsumed under Sydenham's chorea, thus expanding the diagnostic boundaries of Sydenham's chorea to include primarily neuropsychiatric presentations now classified as cases of obsessive-compulsive disorder or Tourette's syndrome. We conclude that PANDAS is a useful construct, but that it would be premature to view it as a subset of Sydenham's chorea-whether defined narrowly or broadly. PMID- 10807958 TI - Influence of prepartum chronic ultramild stress on maternal pup care behavior in mice. AB - BACKGROUND: Stress administered to pregnant rodents has been shown to lead to biological and behavioral alterations in both mother and pups. Most of these stress procedures use noxious stressors. Chronicity is obtained by simple repetition of one or two stressors and may be more representative of moderate daily stress experienced during normal life than stress during severe life events. The effects of this procedure were assessed by observing maternal pup care behavior and testing maternal aggression. METHODS: The subjects included eight controls and eight stressed B6D2F1 females. Chronic ultramild stress was applied from mating to postpartum day (PD) 0. Pup-care behavior was observed on PD 1. Maternal aggression against a male intruder was tested on PD 8, which corresponds to the peak in the display of this behavior. RESULTS: Prenatal stress did not affect basic pup-care behavior, but dramatically impaired defense behavior designed to protect the pups from an external attacker. CONCLUSIONS: The results suggest that gestating females subjected to chronic ultramild stress suffer from a long-lasting decline in recognition of external distress cues either from a resident intruder and/or their own litter. It is assumed these effects are due to the chronicity of the stress rather than its severity. PMID- 10807959 TI - Environmental enrichment and isolation rearing in the rat: effects on locomotor behavior and startle response plasticity. AB - BACKGROUND: Laboratory rats exhibit behavioral changes that reflect a continuum of early life experience, from isolation-reared to socially reared to enrichment reared conditions. In this study, we further characterize the behavioral effects of isolation, social, and enriched rearing on locomotor activity, patterns of movement and exploration, startle reactivity, prepulse inhibition (PPI), and habituation in adult rats. METHODS: Male Sprague-Dawley rat pups (21 days old) were housed under enrichment (three per cage with toys and exposure to enriched environments), normal social (three per cage), or isolation (one per cage) conditions. Eight weeks later, locomotor and exploratory behaviors, acoustic startle reactivity, PPI, and habituation were measured in the three groups. RESULTS: Enrichment-reared rats exhibited reduced exploration and rapid habituation of locomotor activity, increased startle reactivity, and normal PPI and startle habituation compared with socially reared controls. Isolation-reared rats exhibited increased exploration and normal habituation of locomotor activity, increased startle reactivity, reduced PPI, and normal startle habituation. CONCLUSIONS: Isolation- and enrichment-reared rats exhibited opposite changes in some behaviors and similar changes in other behaviors. Specifically, rats raised in enriched conditions appear more efficient at assimilating stimuli from their environment than do rats reared in isolation. Nevertheless, both enrichment- and isolation-rearing conditions increased startle reactivity, whereas only isolation rearing led to disruptions of PPI in adulthood. These results suggest that isolation- and enrichment-rearing conditions produce some common and some differential effects on how rats process environmental stimuli. For studies of isolation-rearing effects on PPI, however, the complex and resource-intensive enrichment condition seems to offer few advantages over the normal social condition. PMID- 10807960 TI - Moxonidine, a selective imidazoline-1 receptor agonist, suppresses the effects of ethanol withdrawal on the acoustic startle response in rats. AB - BACKGROUND: There is a need for improved treatments for ethanol withdrawal in humans. Previously, ethanol withdrawal has been shown to enhance the acoustic startle response in rats. Because many ethanol withdrawal symptoms are caused by autonomic hyperactivity, we examined the effects of two antihypertensives, the imidazoline(I)(1) agonist moxonidine and the alpha(2)-adrenergic partial agonist clonidine, on the ethanol-withdrawal-enhanced acoustic startle response in rats. d-amphetamine-enhanced startle served as a positive control. METHODS: Male, Long Evans rats were made ethanol-dependent through unlimited access to liquid diet containing 6.7% v/v ethanol for 10 days. The concentration of ethanol was reduced to 3.3% v/v on the 11th day. On the 12th day, the rats received control diet. The acoustic startle response was tested 24 hours following the withdrawal of ethanol. Control rats were maintained on control liquid diet throughout the experiment. RESULTS: As has been shown previously, withdrawal from the chronic ingestion of ethanol significantly enhanced the acoustic startle response. Pretreatment with moxonidine (0.01, 0.1, and 1.0 mg/kg, subcutaneously), but not clonidine (0.3, 1.0, and 3.0 mg/kg, subcutaneously), significantly attenuated the ethanol withdrawal-induced elevation of the acoustic startle response. Moxonidine did not suppress the elevation in the startle response caused by d-amphetamine. CONCLUSIONS: These results indicate that I(1) receptors can play an important role in ethanol withdrawal and that moxonidine may be useful for the treatment of ethanol withdrawal in humans. PMID- 10807961 TI - Stimulus novelty differentially affects attentional allocation in PTSD. AB - BACKGROUND: This study investigated attentional allocation in 39 Vietnam combat veterans, 25 with and 14 without posttraumatic stress disorder, assessing P300 amplitudes and latencies during both three-tone and novelty "oddball" tasks. METHODS: The three-tone oddball task consisted of three stimuli: frequent tones (85%), rare target tones (7.5%), and rare distractor tones (7.5%). The novelty oddball task was identical to the three-tone task except that the rare distractor tones were replaced with nonrepeating novel sounds (7.5%). RESULTS: Combat veterans with posttraumatic stress disorder showed significant P300 amplitude enhancements at frontal sites in response to distracting stimuli during the novelty but not during the three-tone oddball tasks. There were no amplitude differences in target tones during either task. CONCLUSIONS: The data suggest that combat veterans with posttraumatic stress disorder demonstrate P300 responses consistent with a heightened orientation response to novel, distracting stimuli. This finding is consistent both with the clinical presentation of the disorder and with theoretical notions that individuals with posttraumatic stress disorder demonstrate information-processing biases towards vague or potentially threatening stimuli. PMID- 10807962 TI - Hormone profiles in humans experiencing military survival training. AB - BACKGROUND: Clinical models of the human response to intense, acute stress have been limited to laboratory settings or cross sectional characterizations. As a result, data about the sensitivity of the human neuroendocrine activation to realistic stressors of varying magnitudes are limited. The U.S. Army survival course offers a unique opportunity to examine, in a controlled manner, the human response to acute, realistic, military stress. METHODS: Salivary data were collected in 109 subjects at baseline during four stress exposure time points and at recovery. Serum data was collected at baseline and recovery in 72 subjects and at baseline and during stress exposure in a subgroup of subjects (n = 21). RESULTS: Cortisol significantly increased during the captivity experience and was greatest after subjects' exposure to interrogations. Cortisol remained significantly elevated at recovery. Testosterone was significantly reduced within 12 hours of captivity. Reductions of both total and free T4 and of total and free T3 were observed, as were increases in thyrotropin. CONCLUSIONS: The stress of military survival training produced dramatic alterations in cortisol, percent free cortisol, testosterone, and thyroid indices. Different types of stressors had varying effects on the neuroendocrine indices. The degree of neuroendocrine changes observed may have significant implications for subsequent responses to stress. PMID- 10807963 TI - Plasma neuropeptide-Y concentrations in humans exposed to military survival training. AB - BACKGROUND: Neuropeptide-Y (NPY) is present in extensive neuronal systems of the brain and is present in high concentrations in cell bodies and terminals in the amygdala. Preclinical studies have shown that injections of NPY into the central nucleus of the amygdala function as a central anxiolytic and buffer against the effects of stress. The objective of this study was to assess plasma NPY immunoreactivity in healthy soldiers participating in high intensity military training at the U.S. Army survival school. The Army survival school provides a means of observing individuals under high levels of physical, environmental, and psychological stress, and consequently is considered a reasonable analogue to stress incurred as a result of war or other catastrophic experiences. METHODS: Plasma levels of NPY were assessed at baseline (prior to initiation of training), and 24 hours after the conclusion of survival training in 49 subjects, and at baseline and during the Prisoner of War (P.O.W.) experience (immediately after exposure to a military interrogation) in 21 additional subjects. RESULTS: Plasma NPY levels were significantly increased compared to baseline following interrogations and were significantly higher in Special Forces soldiers, compared to non-Special Forces soldiers. NPY elicited by interrogation stress was significantly correlated to the subjects' behavior during interrogations and tended to be negatively correlated to symptoms of reported dissociation. Twenty four hours after the conclusion of survival training, NPY had returned to baseline in Special Forces soldiers, but remained significantly lower than baseline values in non-Special Forces soldiers. NPY was positively correlated with both cortisol and behavioral performance under stress. NPY was negatively related to psychological symptoms of dissociation. CONCLUSIONS: These results provide evidence that uncontrollable stress significantly increases plasma NPY in humans, and when extended, produces a significant depletion of plasma NPY. Stress induced alterations of plasma NPY were significantly different in Special Forces soldiers compared to non-Special Forces soldiers. These data support the idea that NPY may be involved in the enhanced stress resilience seen in humans. PMID- 10807964 TI - In humans, serum polyunsaturated fatty acid levels predict the response of proinflammatory cytokines to psychologic stress. AB - BACKGROUND: Psychologic stress in humans induces the production of proinflammatory cytokines, such as interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6), and that of the negative immunoregulatory cytokine, IL-10. An imbalance of omega6 to omega3 polyunsaturated fatty acids (PUFAs) in the peripheral blood causes an overproduction of proinflammatory cytokines. The omega3 PUFAs reduce the production of proinflammatory cytokines. METHODS: This study examines whether an imbalance in omega6 to omega3 PUFAs in human blood predicts a greater production of proinflammatory cytokines in response to psychologic stress. Twenty-seven university students had serum sampled a few weeks before and after as well as 1 day before a difficult oral examination. We determined the omega6 and omega3 fractions in serum phospholipids as well as the ex vivo production of IFN-gamma, TNF-alpha, IL-6, IL-10, and IL-5 by diluted whole blood stimulated with polyclonal activators. RESULTS: Academic examination stress significantly increased the ex vivo, stimulated production of IFN-gamma, TNF-alpha and IL-10, and the IFN-gamma/IL-5 production ratio. Subjects with lower serum omega3 PUFA levels or with a higher omega6/omega3 ratio had significantly greater stress induced TNF-alpha and IFN-gamma responses than subjects with higher serum omega3 PUFAs and a lower omega6/omega3 ratio, respectively. Subjects with lower serum omega3 PUFA levels or with a higher omega6/omega3 ratio had a significantly higher stress-induced increase in the IFN-gamma/IL-5 ratio than the remaining subjects. CONCLUSIONS: Psychologic stress induces a Th-1-like or proinflammatory response in some subjects. An imbalance in the omega6 to omega3 PUFA ratio appears to predispose humans toward an exaggerated Th-1-like response and an increased production of monocytic cytokines, such as TNF-alpha, in response to psychologic stress. The results suggest that increased omega3 PUFA levels may attenuate the proinflammatory response to psychologic stress. PMID- 10807965 TI - Sleep duration, illumination, and activity patterns in a population sample: effects of gender and ethnicity. AB - BACKGROUND: Current knowledge of the population's sleep durations emanates primarily from questionnaires and laboratory studies. Using Actillumes, we investigated whether self-reported sleep durations were indicative of a population decline in sleep duration. We also explored illumination and activity patterns. METHODS: San Diego adults (n = 273, age range: 40-64) were recruited through random telephone calls and were monitored at home while engaging in usual daily routines. RESULTS: Volunteers slept an average of 6.22 hours and received an average of 554 lux (environmental illumination). The timing of sleep, illumination, and activity occurred at 2:44, 12:57, and 13:43, respectively. Irrespective of ethnicity, age, and time reference, men received greater illumination than did women, but this gender effect was not independent of work status. Women and men exhibited a similar circadian activity profile; however, women exhibited better sleep-wake patterns. Interactions between gender and ethnicity suggested worse sleep-wake patterns among minority men. An age-related decline in activity was found, but no age trend in sleep duration or illumination patterns was observed. CONCLUSIONS: This study showed an objective population decline in sleep duration. Sociodemographic effects should be considered in analyses of sleep-wake patterns and illumination exposures. PMID- 10807966 TI - Prefrontal and medial temporal correlates of repetitive violence to self and others. AB - BACKGROUND: The neurobiological basis for violence in humans is poorly understood, yet violent behavior (to self or others) is associated with large social and healthcare costs in some groups of patients (e.g., the mentally retarded). The prefrontal cortex and amygdalo-hippocampal complex (AHC) are implicated in the control aggression, therefore we examined the neural integrity of these regions in violent patients with mild mental retardation and nonviolent control subjects. METHODS: We used (1)H-magnetic resonance spectroscopy (MRS) to measure 1) concentrations and ratios of N-acetyl aspartate (NAA), creatine phosphocreatine (Cr+PCr), and choline-related compounds (Cho) in prefrontal lobe of 10 violent inpatients and 8 control subjects; 2) ratios of NAA, Cr+PCr, and Cho in the AHC of 13 inpatients and 14 control subjects; and 3) frequency and severity of violence in patients. RESULTS: Compared to control subjects, violent patients had significantly (p <.05, analysis of covariance-age and IQ as confounding covariates) lower prefrontal concentrations of NAA and Cr+PCr, and a lower ratio of NAA/Cr+PCr in the AHC. Within the violent patient group, frequency of observed violence to others correlated significantly with prefrontal lobe NAA concentration (r = -0.72, p <.05). CONCLUSIONS: NAA concentration indicates neuronal density, and Cr+PCr concentration high-energy phosphate metabolism. Our findings suggest that violent patients with mild mental retardation have reduced neuronal density, and abnormal phosphate metabolism in prefrontal lobe and AHC compared to nonviolent control subjects. Further studies are needed, however, to determine if these findings are regionally specific, or generalize to other groups of violent individuals. PMID- 10807967 TI - Chemical and biological activity of free radical 'scavengers' in allergic diseases. AB - Reactive oxygen species (ROS) are generated constantly in vivo. They can lead to lipid peroxidation and oxidation of some enzymes, as well as protein oxidation and degradation. Cells possess several biological systems, defined as 'scavengers', to protect themselves from the radical-mediated damage. Immune cells may discharge their arsenal of toxic agents against host tissues, resulting in oxidative damage and inflammation. Therefore, free radical production and disturbance in redox status can modulate the expression of a variety of immune and inflammatory molecules, leading to inflammatory processes, both exacerbating inflammation and effecting tissue damage. Recently, abnormal immunity has been related to oxidative imbalance, and antioxidant functions are linked to anti inflammatory and/or immunosuppressive properties. Currently, allergy is one of the most important human diseases. We studied the role of the primary antioxidant defence system, constituted by the antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase, protecting cells from toxic oxygen. We analyzed how they are involved in blood cells detoxification, and how the imbalance of reactive oxygen species is related to inflammation in allergic diseases by affecting immune cells. Finally, we discuss the published data that relates anti-free radical therapy to the management of human allergic diseases. PMID- 10807968 TI - Effect of hyperthermia on the viability and the fibrinolytic potential of human cancer cell lines. AB - The effects of heat treatment on the viability and fibrinolytic potential of four cultured human carcinoma cell lines, fibrosarcoma cells (HT-1080), lung adenocarcinoma cells with highly metastatic potential (HAL-8), melanoma cells (Bowes) and osteosarcoma cells (NY), determined by measuring their levels of urokinase-type plasminogen activator (u-PA) and its specific receptor (u-PAR), were investigated by comparing them with those of human umbilical vein endothelial cells (HUVECs). HUVECs incubated at 43 degrees C for 120 min exhibited no decrease in viability but exhibited an increase in both u-PA and u PAR. HT-1080 and HAL-8 showed a moderately high heat-resistance (viability, 60 90%) that correlated with the reduction of u-PAR but not u-PA. On the other hand, Bowes and NY cells, with poor heat-resistance (viability, 20-50%), exhibited stronger cell-associated u-PA activity when they survived at 43 degrees C for 120 min. Since the u-PA/u-PAR system is directly involved in the invasiveness and metastatic potential of carcinoma cells, hyperthermia would alter the biological activity of these carcinoma cells. PMID- 10807969 TI - Limitations of urinary telomerase activity measurement in urothelial cancer. AB - The reported frequency of detectable telomerase activity in spontaneously voided urine samples from patients with urothelial cancer varied from 0 to 85%. We examined stasis in the bladder and specimen storage as interfering conditions in this assay. Telomerase activity in exfoliated cells was measured by a polymerase chain-reaction-based assay in spontaneously voided urine from urothelial cancer patients. Effects of retention in the bladder and specimen storage from voiding to measurement of telomerase activity were modeled by suspending 10(6) cells from the cancer-derived T24 line in normal urine (pH 6.5) at 37 degrees C and 25 degrees C, respectively. Hematuria was modeled by adding hemoglobin. In T24 cells suspended in urine at 37 degrees C, telomerase activity had decreased to approximately 20% of preincubation activity after 1 h, and had disappeared after 3 h. In urine at 25 degrees C, telomerase activity in T24 cells had decreased to approximately 40% of preincubation activity at 1 h and to <10% at 6 h. When we examined telomerase activity in exfoliated cells in spontaneously voided urine from urothelial cancer patients (excluding first-voided morning specimens), telomerase activity was detected in only 21% of samples (four of 19) despite measurement with 1 h of voiding and steps to avoid hemoglobin interference. Measurement of telomerase activity in spontaneously voided urine is insufficiently sensitive and reliable for the diagnosis of urothelial cancer. PMID- 10807970 TI - One-step enzyme-linked immunosorbent assay (ELISA) for measurement of serum free leptin. AB - In man, circulating leptin levels are increased with obesity and are regulated by a complex of hormonal, feeding and body-weight changes. Accurate and precise methods to quantitate circulating serum free leptin (f-leptin) concentrations are needed for physiological and clinical studies. We developed a one-step enzyme immunoassay to measure human f-leptin in serum. The detection limit was 0.40 ng/ml. The recovery of leptin added to serum was 90.8-102.8%. The within-run and between-day coefficients of variation (C.V.) ranged from 2.8 to 7.7 and 5.7 to 9.7%, respectively, and the immunoassay had an overall recovery rate for serial dilution in the range of 94. 0-109.9%. Measured serum f-leptin concentrations in 201 adults correlated (r=0.449, P<0.001) directly with body mass index (BMI kg/m(2)), particularly when results were separated by gender (r=0. 709 for male, P<0.001; r=0.643 for female, P<0.001). We conclude that this one-step enzyme immunoassay is accurate for measuring f-leptin in human serum. PMID- 10807971 TI - The proteins and the formation of gallstones. AB - Cholesterol supersaturation of bile requires assistance to form gallstones. Proteins have been proposed as candidates either to facilitate or hinder the formation of stones. It is assumed that the identity of these stone proteins should be revealed in order to assess their role in the process. We have used electrodialysis of crushed stones to extract the proteins followed by 2D electrophoresis and N-terminal amino acid sequencing to characterize them. The discovery of bacterial proteins in some stones adds evidence to the importance of an inflammatory process and the deconjugation of bilirubin in mixed gallstone formation. PMID- 10807972 TI - Comparison of representative ranges based on U.S. patient population and literature reference intervals for urinary trace elements. AB - Reference intervals for trace elements are very hard to obtain because of the difficulty of defining a nonexposed reference population. However, representative ranges for trace elements obtained from a general patient population can provide useful information in interpreting laboratory results. We have used urine specimens submitted for trace metal analysis from patients residing in the United States to calculate representative ranges for 25 urinary trace elements, and to compare them to reference values taken from the literature. All urine analytes were measured by inductively-coupled plasma-mass spectrometry except chromium, which was measured by graphite furnace atomic absorption spectroscopy. For representative range calculation two approaches were used. In the non-parametric calculation first, the top 10% of results were discarded assuming that those specimens came from individuals with unusually high trace element exposures. Next the central 95% of the remaining data was taken as the reference interval. In the parametric calculation the specimens from exposed or not healthy individuals were assumed to appear as outliers and were discarded. The mean and S.D. were calculated, and used to determine representative ranges. The two approaches yielded very similar results, and worked remarkably well for 14 analytes. There were minor discrepancies for 7 analytes, and major for 4 analytes. All analyses of urinary trace elements included a urine creatinine value, which was used to express urinary trace element concentrations in terms of creatinine ratio. This corrects for differences in urine concentration that affects the results for random specimens. PMID- 10807973 TI - Characteristics of L-ornithine: 2-oxoacid aminotransferase and potential prenatal diagnosis of gyrate atrophy of the choroid and retina by first trimester chorionic villus sampling. AB - A deficiency of the mitochondrial matrix enzyme L-ornithine: 2-oxoacid aminotransferase causes gyrate atrophy of the choroid and retina with hyperornithinemia (MIM 258870), a blinding degenerative disease, which is inherited as an autosomal recessive trait. We have developed a sensitive microradioisotopic method for enzyme assay by using 2-oxo-[5-14C] glutarate as the substrate and performing the separation of the product, [5-14C] glutamate from the substrate on a cation-exchange column. The enzyme activity was determined in human and rat tissues and in cultured cells. The enzyme activity in fibroblasts from a patient was deficient and that of the parents ranged between 25 and 60% of the control values. In addition we have found the enzyme expressed in native and cultured chorionic villi indicating a potential detection of the disease during the first trimester of pregnancy. PMID- 10807974 TI - The cytochrome P450 2D6 (CYP2D6) gene polymorphism among breast and head and neck cancer patients. AB - The prevalence of CYP2D6*3 and CYP2D6*4 alleles in normal controls and cancer patients was studied using the reliable PCR-SSCP method. In the control group (n=144), four subjects (2.8%) were found to carry CYP2D6*3 allele (heterozygote), while 30 (20.8%) subjects carried CYP2D6*4 allele (18.8% heterozygotes, 2.1% homozygotes). One (1.3%) of the breast cancer (BC) patients (n=76) carried CYP2D6*3 allele, but 24 (31.6%) carried CYP2D6*4 allele (26.3% heterozygotes, 5.3% homozygotes). In the head and neck cancer (HNC) group (n=56), two (3.6%) patients were heterozygous for CYP2D6*3 mutation and 15 (26.8%) for CYP2D6*4 mutation. Fourteen of 56 (25%) and one of 56 (1. 8%) of these patients carried heterozygous and homozygous mutations, respectively. In controls, 2.1% were identified as poor metabolizers (PM), 76.4% as extensive metabolizers (EM), and 21.5% as intermediate heterozygotes (IEM). In BC group, 5.3, 27.6 and 67.1% were classified as PM, IEM and EM, respectively. In HNC group, the incidence of PM was 1.8, but as many as 28.6% were identified as IEM phenotypes. PMID- 10807975 TI - Preliminary report of the clinical performance of a new urinary bladder cancer antigen test: comparison to voided urine cytology in the detection of transitional cell carcinoma of the bladder. AB - We compared the ability of a new urinary bladder cancer antigen (UBC) test with conventional cytology for the detection of transitional cell carcinoma of the bladder using voided urine samples. The UBC was measured and corrected for the creatinine concentration in the urine of 61 patients with transitional cell carcinoma of the bladder (group 1), 23 patients without recurrent bladder tumors during follow-up (group 2), 28 patients with benign prostatic hyperplasia (group 3), nine patients with prostate cancer (group 4), and 90 healthy volunteers free of urological diseases (group 5). The UBC concentrations were 408.8+/-578.5, 18.8+/-26.6, 23.9+/-32.7, 17.5+/-18.6 and 4.6+/-6.7 ngmg(-1) creatinine (mean+/ S. D.) for groups 1-5, respectively. The level for group 1 was significantly higher than for any other group. The sensitivity and specificity, which were optimized using receiver-operating characteristic curves for groups 1 and 2 were 82.0% and 82.6%, respectively, at a threshold value of 39 ngmg(-1) creatinine. The sensitivity and specificity of cytology for these same groups were 60.7% and 86.9%, respectively. The sensitivity of the UBC was significantly higher than that of cytology, not only for total bladder tumors (82.0% vs. 60.7%, P<0.02) but also for grade I transitional cell carcinoma of the bladder (76.5% vs. 11.8%, P<0. 001). While offering a similarly high specificity, the UBC test might have an advantage over cytology in terms of superior sensitivity, particularly for low grade tumors. PMID- 10807976 TI - Oxidized lipoprotein(a) increases the expression of platelet-derived growth factor-B in human umbilical vein endothelial cells. AB - Lipoprotein(a) ?Lp(a) has been demonstrated to be an independent risk factor for atherosclerosis and restenosis after angioplasty. However, the precise mechanism by which it contributes to the development of both remains unclear. Platelet derived growth factor (PDGF) is one of the key factors that induce the proliferation and migration of vascular smooth muscle cells. The present study investigated the effects of native and oxidized LDL ?n-LDL and ox-LDL and Lp(a) ?n-Lp(a) and ox-Lp(a) on the expression of PDGF-B in cultured human umbilical vein endothelial cells (HUVECs). Results showed that PDGF-B expression was not influenced by n-LDL, but was moderately increased by ox-LDL and n-Lp(a). Ox-Lp(a) was the most potent stimulus for PDGF-B expression, increasing it in HUVECs by 156%+/-18% at 5 nmol/l and 219%+/-42% at 20 nmol/l. Northern Blot analysis demonstrated that the amount of PDGF-B mRNA was markedly increased after treatment with ox-Lp(a) but not n-LDL, ox-LDL and n-Lp(a). These results demonstrate that ox-Lp(a) can elicit PDGF-B expression in HUVECs, which may thereby influence the pathogenesis of atherosclerosis and restenosis after angioplasty. PMID- 10807977 TI - Relation between RLP-triglyceride to RLP-cholesterol ratio and particle size distribution in RLP-cholesterol profiles by HPLC. AB - Remnant-Like Particles (RLP) isolated by an immunoseparation method are heterogeneous in their physical and biochemical properties. The objective of this study was to examine the relation between RLP-triglyceride (RLP-TG) to RLP cholesterol (RLP-C) ratio and particle size distribution in RLP-C profiles from patients with hyperlipoproteinemia by HPLC. RLP were isolated from serum samples from 147 subjects. RLP-C and RLP-TG were quantified by respective enzymatic methods. Particle sizes of the RLP were measured using HPLC with 4 connected TSKgel LipopropakXL columns. Based on HPLC profiles of RLP-C from individual subjects, three different types were classified: predominantly LDL, predominantly VLDL, and mostly VLDL types. All patients with type III hyperlipidemia were mostly VLDL type but with smaller particle size of VLDL (32 nm) than other subjects. Severe hypertriglyceridemic (TG>4.52 mmoll(-1)) subjects were mostly VLDL type with large particle size (41 nm). As for all subjects (n=105) without predominantly LDL type, a significant correlation between RLP particle size and RLP-TG to RLP-C ratio (r=0. 432, P<0.001) was obtained, but not in case of serum TG to RLP-C ratio (r=0.062). It suggests that RLP-TG to RLP-C ratio might be used for discrimination of atherogenic smaller-sized lipoprotein from larger-sized TG rich lipoprotein remnants. PMID- 10807978 TI - Spectrophotometric assay for serum platelet-activating factor acetylhydrolase activity. AB - We developed a spectrophotometric assay for serum platelet-activating factor acetylhydrolase (PAF-AH, EC 3.1.1.47.) activity using a platelet-activating factor (PAF) analogue with a 4-nitrophenyl group as substrate. PAF-AH hydrolyzes the sn-2 position of the substrate ?1-myristoyl-2-(p nitrophenylsuccinyl)phosphatidylcholine, producing p-nitrophenyl succinate. This liberation was spectrophotometrically monitored and the activity determined from the change in absorption. The assay does not require radioisotopes and is applicable to an automatic analyzer. Utilizing this assay with an automatic analyzer, it is possible to measure the activities of thousands of samples in a few hours with excellent precision (CV 0.5%, n=30) and high correlation (r=0.979, n=100) with the results of a conventional radioisotopic assay. The assay should be particularly useful for clinical diagnostics. PMID- 10807979 TI - Reference range of serum haptoglobin is haptoglobin phenotype-dependent in blacks. AB - Reference values for serum haptoglobin (Hp), were established in a Black Zimbabwean population. The upper limit (2.15 g/l) is comparable to the one in Caucasians, but the lower limit (0.12 g/l) is much lower than the proposed interim international reference limit (0.3 g/l). Subjects that typed as Hp 0-0 by starch gel electrophoresis technique were retyped using high performance gel permeation chromatography. This resulted in a 32% decrease in the frequency of Hp 0-0, but an increase in Hp 2-2 and Hp 2-1M phenotype frequencies. In the Zimbabwean Blacks, the Hp 0-0 frequency was estimated to be 2.9%. Haptoglobin reference values were found to be Hp phenotype-dependent; highest values were found in Hp 1-1 (median 0.88 g/l; range 0.31-1.69 g/l) and in Hp 2-1 (median 0.90 g/l; range 0.31-2.22 g/l) and lower values (median 0.66 g/l; range 0.13-1.79 g/l) in Hp 2-2 subjects. The Hp 2-1M phenotype was characterized by low reference values (0.18-1.25 g/l) (P<0.05). In three cases of the rare variant Hp Johnson, high Hp concentrations were found (median 1. 57 g/l; range 0.98-1.57 g/l). PMID- 10807980 TI - Immunochemical characterization of monoclonal protein in the serum of a patient with Waldenstr]om's macroglobulinemia showing both pyroglobulin and cryoglobulin properties. AB - A patient with Waldenstr]om's macroglobulinemia whose serum demonstrated properties of both pyroglobulin and cryoglobulin was studied. A monoclonal (M) protein in the serum of the patient was identified by immunoelectrophoresis (IEP) and immunofixation electrophoresis (IFE) as IgM-lambda. The M-protein was separated by gel permeation high-performance liquid chromatography (HPLC). Cryoglobulin in the serum was isolated by the method of cold precipitation. The cryoglobulin was identified as IgM-lambda type by IEP and was the same M-protein as that which occurred in this patient's serum. The purified cryoglobulin also had the properties of a pyroglobulin. Neither property disappeared following pretreatment with 2-mercaptoethanol (2-ME), urea, and Triton X-100 detergent, and deglycosylation with N-glycanase (E.C. 3.5.1.52). We suggest that these abnormal properties were caused by the molecular abnormality of the IgM-lambda M-protein. PMID- 10807981 TI - Plasma nitrite/nitrate concentrations as a tumor marker for hepatocellular carcinoma. AB - Since plasma concentrations of nitrite/nitrate, the stable end-products of nitric oxide, increase in patients with hepatocellular carcinoma (HCC) correlatively to tumor volume, we examined the ability of plasma nitrite/nitrate to discriminate between those patients with HCC and those without and compared the diagnostic performance of the parameter with that of serum alpha-fetoprotein (AFP) concentrations. Plasma nitrite/nitrate and serum AFP concentrations were measured using a Griess reaction and a solid phase enzyme immunoassay, respectively. Eighty-nine patients with chronic liver diseases (CLD) with (n=39) or without HCC (n=50) and 50 healthy control subjects participated in the study. A receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value and accuracy. The areas under ROC curves for nitrite/nitrate and AFP were calculated to be 0.758 and 0.812, respectively, which were not significantly different. There was no correlation between the concentrations of plasma nitrite/nitrate and serum AFP. The sensitivity, the specificity, and diagnostic efficiency were 79.5, 72.0, and 75.3%, respectively, for nitrite/nitrate, and 74.4, 76.0, and 75.3%, respectively, for AFP. Based on a partial ROC curve, the clinical utility of plasma nitrite/nitrate as a tumor marker approximated that of serum AFP, but exceeded in AFP-negative patients. Indeed, nitrite/nitrate was positive in 70% of AFP-negative HCC patients. The simultaneous determinations of serum AFP and plasma nitrite/nitrate concentrations gave significant improvement in detection of HCC in CLD patients compared with that of serum AFP alone. PMID- 10807982 TI - Heterologous enzyme immunoassay for serum androstenediol. AB - A heterologous enzyme immunoassay for serum androstenediol (Adiol: 3beta, 17beta dihydroxy-androst-5-ene) was established. The combination of anti-Adiol antiserum raised in rabbit against Adiol 7-O-(carboxymethyl)oxime (Adiol 7-CMO) conjugated bovine serum albumin (Adiol 7-CMO-BSA) and Adiol 7-iminomethylcarboxylic acid conjugated alkaline phosphatase was used for the assay. The sensitivity of the heterologous assay system was superior to that of a homologous assay system in which an antibody raised in rabbit against Adiol 7-CMO-BSA and enzyme labeled antigen, Adiol 7-CMO conjugated alkaline phosphatase, were used. The minimal amount of Adiol detected was 0.4 ngml(-1) and the measurable range was from 0. 4 to 150 ngml(-1). Intra-assay coefficients of variation (C.V.) were 8.6% (1.52+/ 0.13 ngml(-1), mean+/-S.D., n=10) and 6.7% (13.4+/-0.9 ngml(-1), n=10). Inter assay C.V. were 12.9% (1.63+/-0.21 ngml(-1), n=8) and 11.5% (12.2+/-1.4 ngml(-1), n=8). A linear relation was observed between the serum sample dilution and the Adiol concentration. For recovery study, authentic Adiol was added to serum sample (original concentration: 1.43 ngml(-1)). The calculated final Adiol concentration was 2.99 ngml(-1). The recovery was 98.6% (n=5). The Adiol concentrations in healthy subjects measured by the proposed assay (male: 1.1+/ 0.3 ngml(-1) (mean+/-S.D.), range: 0.7-1. 7 ngml(-1), age: 22-50, n=10; female: 0.6+/-0.4 ngml(-1), range: 0. 2-1.6 ngml(-1), age: 23-48, n=20) were consistent with reported values. PMID- 10807983 TI - Role of antioxidant enzymes in brain tumours. AB - Erythrocyte antioxidant enzymes were analysed in 100 patients with intracranial neoplasm and in 47 controls. There was a significant decrease in RBC glutathione reductase (GRx) and superoxide dismutase (SOD) activity in most types of brain tumor cases. Patients with acoustic neurinoma showed a significant reduction in selenium-dependent glutathione peroxidase (Se-GPx) activity. A decrease in catalase (CT) activity was seen in most of the brain tumor patients but remained statistically insignificant when compared to controls. A significant increase in plasma ceruloplasmin concentration was observed in patients with glioma. These enzymes were also studied in 27 post-treatment cases. GRx activity returned to normal levels in these patients. RBC SOD and plasma ceruloplasmin levels showed a tendency to return to normal. Hence, a marked decrease in the antioxidant enzymes may have a role in the genesis of considerable oxidative stress in patients with brain tumors. PMID- 10807984 TI - Finite element modeling of human skin using an isotropic, nonlinear elastic constitutive model. AB - The collagen network in skin is largely responsible for the nonlinear mechanical stress-strain response of skin. We hypothesize that the force-stretch response of collagen is governed by the entropics of long-chain molecules. We show that a constitutive model derived from the statistical mechanics of long-chain molecules, corresponding to the fibrous collagen network in skin, captures the mechanical response of skin. A connection between the physiologically meaningful parameters of network molecular chain density and free length of collagen fibers and the constitutively significant parameters of initial modulus and limiting stretch is thus established. The relevant constitutive law is shown to have predictive capabilities related to skin histology by replicating in vivo and in vitro experimental results. From finite element simulations, this modeling approach predicts that the collagen network in hypertrophic scars is more dense and the constituent collagen fibers have shorter free lengths than in healthy skin. Additionally, the model is shown to predict that as rat skin ages, collagen network density increases and fiber free length decreases. The importance of knowledge of the in situ stress state for analyzing skin response and validating constitutive laws is also demonstrated. PMID- 10807985 TI - A synthetic fiber-reinforced stentless heart valve. AB - There is strong evidence that failure of bioprosthetic and synthetic valves occurs as a consequence of high tensile and bending stresses, acting on the leaflets during opening and closing. In stented prostheses, whether synthetic or biological, the absence of contraction of the aortic base causes the leaflets to be subjected to an unphysiological degree of flexure, which is also related to calcification. However, a stentless synthetic valve, which has a flexible aorta base, can be a good alternative for stented synthetic valves. Moreover, fiber reinforcement is assumed to lead to a decrease of tears and perforation as a result of reduced stresses in the weaker parts of the leaflets in their closed configuration. The manufacturing method for a stentless, fiber-reinforced, synthetic valve is presented. Prototypes are tested in a pulse duplicator system. The results show that the mean systolic pressure difference is very low, while the high regurgitation (up to 26%) is probably caused by a too small coaptation area of the leaflets. PMID- 10807986 TI - The relationship between force depression following shortening and mechanical work in skeletal muscle. AB - Force depression following muscle shortening was investigated in cat soleus (n=6) at 37 degrees C for a variety of contractile conditions with the aim to test the hypotheses that force depression was independent of the speed of shortening and was directly related to the mechanical work produced by the muscle during shortening. Force depression was similar for tests in which the mechanical work performed by the muscle was similar, independent of the speed of shortening (range of speeds: 4-256mm/s). On the other hand, force depression varied significantly at a given speed of shortening but different amounts of mechanical work, supporting the hypothesis that force depression was not speed - but work dependent. The variations in the mechanical work produced by the muscle during shortening accounted for 87-96% of the variance observed in the force depression following shortening further supporting the idea that the single scalar variable work accounts for most of the observed loss in isometric force after shortening. The results of the present study are also in agreement with the notion that the mechanism underlying force depression might be associated with an inhibition of cross-bridge attachments in the overlap zone formed during the shortening phase, as proposed previously (Herzog and Leonard, 1997. Journal of Bimechanics 30 (9), 865-872; Marechal and Plaghki, 1979. PMID- 10807987 TI - Topographical variation of the elastic properties of articular cartilage in the canine knee. AB - Equilibrium response of articular cartilage to indentation loading is controlled by the thickness (h) and elastic properties (shear modulus, mu, and Poisson's ratio, nu) of the tissue. In this study, we characterized topographical variation of Poisson's ratio of the articular cartilage in the canine knee joint (N=6). Poisson's ratio was measured using a microscopic technique. In this technique, the shape change of the cartilage disk was visualized while the cartilage was immersed in physiological solution and compressed in unconfined geometry. After a constant 5% axial strain, the lateral strain was measured during stress relaxation. At equilibrium, the lateral-to-axial strain ratio indicates the Poisson's ratio of the tissue. Indentation (equilibrium) data from our prior study (Arokoski et al., 1994. International Journal of Sports Medicine 15, 254 260) was re-analyzed using the Poisson's ratio results at the test site to derive values for shear and aggregate moduli. The lowest Poisson's ratio (0.070+/-0.016) located at the patellar surface of femur (FPI) and the highest (0.236+/-0.026) at the medial tibial plateau (TMI). The stiffest cartilage was found at the patellar groove of femur (micro=0.964+/-0.189MPa, H(a)=2.084+/-0. 409MPa) and the softest at the tibial plateaus (micro=0.385+/-0. 062MPa, H(a)=1.113+/-0.141MPa). Comparison of the mechanical results and the biochemical composition of the tissue (Jurvelin et al., 1988. Engineering in Medicine 17, 157-162) at the matched sites of the canine knee joint indicated a negative correlation between the Poisson's ratio and collagen-to-PG content ratio. This is in harmony with our previous findings which suggested that, in unconfined compression, the degree of lateral expansion in different tissue zones is related to collagen-to-PG ratio of the zone. PMID- 10807988 TI - Fluid mechanics of regurgitant jets and calculation of the effective regurgitant orifice in free or complex configurations. AB - The velocity fields of turbulent jets can be described using a single formula which includes two empirical constants: k(core) determining the length of the central core and k(turb) the jet widening. Flow models simulating jet adhesion, confinement and noncircular orifices were studied using laser Doppler anemometer and the modifications of the constants were derived from series of velocity profiles. In circular free jets, k(core) was found equal to 4.1 with a variability of 1.4%. In complex configurations, its variability was equal to 15.2%. For k(turb), the value for free circular jets was of 45.2 with a variability of 6.0% and this variability in complex configurations was significantly higher (30. 1%, p=0.025). The correlation between the actual orifice size and the jet extension was poor (r=0.52). However, the almost constant value of k(core) allowed to define a new algorithm calculating the regurgitant orifice diameter with the use of outlines of the jet image (r=0.89). In conclusion, the fluid mechanics of regurgitant jets is modified in complex configurations but, due to the relative independency of the central core, velocity fields could be used to evaluate the dimensions of the effective regurgitant orifice. PMID- 10807989 TI - Identification of linear viscoelastic constitutive models. AB - Accelerations induce in the brain mechanical stresses that may explain the loss of consciousness feared by fighter pilots. In this study, the brain is modelled as a multi-domain structure and a finite element method is used to identify the constitutive law parameters of each domain and then to analyse the stress level in the brain. The loading and observed strain rates induced by hypergravity seem to indicate a quasi-static behaviour of the brain structure. A general procedure has been developed to characterise the behaviour of a structure including several domains. Each of them is assumed to be isotropic and homogeneous with a linear viscoelastic behaviour. These constitutive laws were identified using only the displacements of several nodes on the envelope discarding the displacements between domains at the interaction surfaces. These interfaces may be buried inside the structure and not connected with the external surface. Two validation examples are proposed to show the reliability and effectiveness of the method. PMID- 10807991 TI - On the monodimensional approach to the estimation of the highest reynolds shear stress in a turbulent flow. AB - The measurement of the Reynolds stress tensor, or at least of some of its components, is a necessary step to assess if the turbulence associated with the flow near prosthetic devices can damage blood constituents. Because of the intrinsic three dimensionality of turbulence, in general, a three-component anemometer should be used to measure directly the components of the Reynolds stress tensor. However, this can be practically unfeasible, especially in vivo; therefore, it is interesting to investigate the possibility of characterizing the turbulent flows that may occur in the circulatory system with the monodimensional data that a less complete equipment (e.g., a pulsed ultrasound Doppler) can yield. From the general expression of the Reynolds stress tensor, the highest shear stress can be deduced, as well as the Reynolds normal stress in the main flow direction. The relation between these two quantities, which is an issue already addressed in previous works, can thus be rigorously formulated in terms of some characteristic parameters of the Reynolds stress tensor, the principal normal stresses and the angles that the directions that define them form with the main flow direction. An experimental verification of the ratio of the two above mentioned quantitites for the flow across bileaflet valves, investigated by means of two-dimensional laser Doppler anemometry, will illustrate the limitations of the monodimensional approach estimating the maximum load on blood constituents. PMID- 10807990 TI - Mechanical effects of continuous passive motion on the lumbar spine in seating. AB - The aim of this study was to develop a model which describes the mechanical spinal response to small alternating pelvic stimulation induced by an active rotational movement of a normal chair. The rotary continuous passive motion (RCPM) of the seat about a vertical axis of only 0.6 degrees resulted in an increased in spinal length as opposed to the normal daily shrinkage, and back patients experienced pain relief. Passive and active exercies have been broadly applied for treating and healing spinal disorders. A rigid body package (ADAMS Android) was used to translate the stimulation of the ischial tubersoity in caudo cranial handing-over visualisation. The parameters of the model were set so that the values of the global stiffness and geometry of the intervertebral discs could be changed. In vivo validation of the model was based on force and moment measurements using an internal AO fixator. The predicitons of the model concerning natural frequency (4.5Hz) in vertical direction and the axial torsion response on small pelvic torsion are comparable with experimental data. PMID- 10807992 TI - Effect of endothelial injury and increased blood pressure on albumin accumulation in the arterial wall: a numerical study. AB - The present study investigates the influence of endothelial damage and of blood pressure on albumin accumulation in the arterial wall. For this purpose a numerical model for the coupled mass transport processes in the arterial lumen and in the various layers of the arterial wall is developed. The model considers the transport in the endothelium, intima, internal elastic lamina (IEL) and media of a straight axisymmetric arterial segment. In the arterial lumen fully developed stationary blood flow is assumed, the filtration velocity in the wall layers is calculated applying Darcy's law. The description of the luminal mass transport uses the stationary convection-diffusion equation, the transport in the porous intima and media is modelled applying the volume-averaged stationary convection-diffusion-reaction equation. The transport processes in the lumen, intima and media are coupled by the flux across the endothelium and IEL, which is mathematically described using the Kedem-Katchalsky equations. The numerical solution of the transport equations applies the finite element method. The results demonstrate a high resistance of the healthy endothelium to macromolecule exchange between blood and the artery wall. The reduced resistance of an injured endothelium causes an increased mass flux into the wall which results in higher concentration levels within the wall. The effect of the blood pressure on the wall concentration level is different for a helathy and an injured endothelium. In the case of a healthy endothelium a blood pressure increase causes a decrease of the intimal concentration and an increase of the medial concentration, whereas in the case of an injured endothelium an increased blood pressure results in higher concentration levels within the intima and media. PMID- 10807993 TI - Numerical study on the effect of secondary flow in the human aorta on local shear stresses in abdominal aortic branches. AB - Flow in the aortic arch is characterized primarily by the presence of a strong secondary flow superimposed over the axial flow, skewed axial velocity profiles and diastolic flow reversals. A significant amount of helical flow has also been observed in the descending aorta of humans and in models. In this study a computational model of the abdominal aorta complete with two sets of outflow arteries was adapted for three-dimensional steady flow simulations. The flow through the model was predicted using the Navier-Stokes equations to study the effect that a rotational component of flow has on the general flow dynamics in this vascular segment. The helical velocity profile introduced at the inlet was developed from magnetic resonance velocity mappings taken from a plane transaxial to the aortic arch. Results showed that flow division ratios increased in the first set of branches and decreased in the second set with the addition of rotational flow. Shear stress varied in magnitude with the addition of rotational flow, but the shear stress distribution did not change. No regions of flow separation were observed in the iliac arteries for either case. Helical flow may have a stabilizing effect on the flow patterns in branches in general, as evidenced by the decreased difference in shear stress between the inner and outer walls in the iliac arteries. PMID- 10807994 TI - Alignment and proliferation of MC3T3-E1 osteoblasts in microgrooved silicone substrata subjected to cyclic stretching. AB - Previous studies have shown that many types of cells align in microgrooves in static cultures. However, whether cells remain aligned and also proliferate in microgrooves under stretching conditions has not been determined. We grew MC3T3 E1 osteoblasts in deformable silicone dishes containing microgrooves oriented in the stretch direction. We found that with or without 4% stretching, cells aligned in microgrooves of all sizes, with the groove and ridge widths ranged from 1 to 6microm, but the same groove depth of about 1.6microm. In addition, actin cytoskeleton and nuclei became highly aligned in the microgrooves with and without 4% cyclic stretching. To further examine whether MC3T3-E1 osteoblasts proliferate in microgrooves with cyclic stretching, we grew the cells in six-well silicone dishes containing microgrooves in three wells and smooth surfaces in other three wells. After 4% cyclic stretching for 3, 4, and 7 days, we found that cell numbers in the microgrooves were not significantly different (p>0.05) from those on the smooth surface (p>0.05). Taken together, these results show that MC3T3-E1 osteoblasts can align and proliferate in microgrooves with 4% cyclic stretching. We suggest that the silicone microgrooves can be a useful tool to study the phenotype of MC3T3-E1 osteoblasts under controlled substrate strains. The silicone microgrooves can also be useful for delivering defined substrate strains to other adherent cells in cultures. PMID- 10807995 TI - Strategies used to stabilize the elbow joint challenged by inverted pendulum loading. AB - The stiffness of activated muscles may stabilize a loaded joint by preventing perturbations from causing large displacements and injuring the joint. Here the elbow muscle recruitment patterns were compared with the forearm loaded vertically (a potentially unstable inverted pendulum configuration) and with horizontal loading. Eighteen healthy subjects were studied with the forearm vertical and supinated and the elbow flexed approximately 90 degrees. In the first experiment EMG electrodes recorded activity of biceps, triceps, and brachioradialis muscles for joint torques produced (a) by voluntarily exerting a horizontal force isometrically (b) by voluntarily flexing and extending the elbow while the forearm was loaded vertically with 135N. The relationship between the EMG and the torque generated was quantified by the linear regression slope and zero-torque intercept. In a second experiment a vertical load increasing linearly with time up to 300N was applied. In experiment 1 the EMG-torque relationships for biceps and triceps had an intercept about 10% of maximum voluntary effort greater with the vertical compared to the horizontal force, the inverse was found for Brachioradialis, but the EMG-torque slopes for both agonist and antagonistic muscles were not different. In experiment 2 there were 29 trials with minimal elbow displacement and all the three muscles activated on the order of 11% of maximum activation to stabilize the elbow; 19 trials had small elbow extension and 14 trials small flexion requiring altered muscle forces for equilibrium; 7 trials ended in large unstable displacement or early termination of the test. An analysis indicate that the observed levels of muscle activation would only provide stability if the muscles' short-range stiffness was at the high end of the published range, hence the elbow was marginally stable. The stability analysis also indicated that the small elbow extension increased stability and flexion decreased stability. PMID- 10807996 TI - Experimental production of extra- and intra-articular fractures of the os calcis. AB - Although studies have been conducted in the past to duplicate traumatic fractures of the os calcis, biomechanical force data as a function of extra- and intra articular fractures are not available. Consequently, in this study, a dynamic single impact model was used to provide such information. Using intact human cadaver lower extremities, impact loading was applied to the plantar surface of the foot using a mini-sled pendulum equipment. The proximal tibia was fixed in polymethylmethacrylate. Following impact, pathology to the os calcis was classified into intact (no injury; 14 cases), and extra-articular (6 cases) and intra-articular (6 cases) fractures. Peak dynamic forces were used to conduct statistical analysis. Mean forces for the intact and (both) fracture groups were 4144 N (standard error, SE: 689) and 7802 N (SE: 597). Mean forces for the extra- and intra-articular fracture groups were 7445 N (SE: 711) and 8159 N (SE: 1006). The peak force influenced injury outcome (ANOVA, p<0.005). Differences in the forces were found between intact and injured specimens (p<0.01); intact specimens and specimens with extra-articular pathology (p<0.001); intact specimens and specimens with intra-articular pathology (p<0.005). The present experimental protocol, which successfully reproduced clinically relevant os calcis pathology, can be extended to accommodate other variables such as the simulation of Achilles tendon force, the inclusion of other angles of force application, and the application of the impact force to limited regions of the plantar force of the foot in order to study other injury mechanisms. PMID- 10807997 TI - Functional assessment in the rat by ground reaction forces. AB - Although the rat sciatic nerve model is used extensively in the investigation of repair techniques, and a variety of evaluation methods utilized to assess the results, a means to measure directly and accurately the return of function in these animals is absent. Histologic, histomorphometric, and electrophysiologic methods can be reliable indicators of nerve regeneration but do not correlate to functional recovery. The purposes of this study were to develop apparatus to continuously measure ground reaction forces (GRF) and use GRF parameters in the assessment of gait parameters in normal rats preoperatively and following peripheral nerve severance and repair. Three neurorrhaphy methods: direct sciatic nerve repair, direct tibial nerve repair and double sciatic nerve repair simulating autograft, as well as a non-repaired tibial nerve transection were evaluated. The testing apparatus was designed to measure the spontaneous and voluntary effort of the rat with objective data. Three orthogonal components - vertical, craniocaudal (braking and propulsion), and mediolateral - of the ground reaction force were measured. Preoperative data showed that vertical forces were comparable among the four limbs but propulsion and braking forces displayed significant differences. At 12 weeks, functional recovery was most evident in the direct tibial nerve repair group and absent in the non-repaired tibial defect group. Direct sciatic nerve repairs and sciatic nerve grafts resulted in lesser degrees of improvement. Results indicated that the propulsive force is the optimal GRF parameter for evaluating recovery of useful function. PMID- 10807998 TI - A single camera roentgen stereophotogrammetry method for static displacement analysis. AB - A new method to quantify motion or deformation of bony structures has been developed, since quantification is often difficult due to overlaying tissue, and the currently used roentgen stereophotogrammetry method requires significant investment. In our method, a single stationary roentgen source is used, as opposed to the usual two, which, in combination with a fixed radiogram cassette holder, forms a camera with constant interior orientation. By rotating the experimental object, it is possible to achieve a sufficient angle between the various viewing directions, enabling photogrammetric calculations. The photogrammetric procedure was performed on digitised radiograms and involved template matching to increase accuracy. Co-ordinates of spherical markers in the head of a bird (Rhea americana), were calculated with an accuracy of 0.12mm. When these co-ordinates were used in a deformation analysis, relocations of about 0.5mm could be accurately determined. PMID- 10807999 TI - A continuous pure moment loading apparatus for biomechanical testing of multi segment spine specimens. AB - An apparatus is described that enables the application of continuous pure moment loads to multi-segment spine specimens. This loading apparatus allows continuous cycling of the spine between specified flexion and extension (or right and left lateral bending) maximum load endpoints. Using a six-degree-of-freedom load cell and three-dimensional optoelectronic stereophotogrammetry, characteristic displacement versus load hysteresis curves can be generated and analyzed for different spinal constructs of interest. Unlike quasi-static loading, the use of continuous loading permits the analysis of the spine's behaviour within the neutral zone. This information is of particular clinical significance given that the instability of a spinal segment is related to its flexibility within the neutral zone. Representative curves for the porcine lumbar spine in flexion extension and lateral bending are presented to illustrate the capabilities of this system. PMID- 10808000 TI - Energy-conserving impact algorithm for the heel-strike phase of gait. AB - Significant ground reaction forces exceeding body weight occur during the heel strike phase of gait. The standard methods of analytical dynamics used to solve the impact problem do not accommodate well the heel-strike collision due to the persistent contact at the front foot and presence of contact at the back foot. These methods can cause a non-physical energy gain on the order of the total kinetic energy of the system at impact. Additionally, these standard techniques do not quantify the contact force, but the impulse over the impact. We present an energy-conserving impact algorithm based on the penalty method to solve for the ground reaction forces during gait. The rigid body assumptions are relaxed and the bodies are allowed to penetrate one another to a small degree. Associated with the deformation is a potential, from which the contact forces are derived. The empirical coefficient-of-restitution used in the standard approaches is replaced by two parameters to characterize the stiffness and the damping of the materials. We solve two simple heel-strike models to illustrate the shortcomings of a standard approach and the suitability of the proposed method for use with gait. PMID- 10808001 TI - Back lift versus leg lift: an index and visualization of dynamic lifting strategies. AB - The description of a lifting strategy is typically provided in qualitative terms. A quantitative static descriptor or index differentiates the starting postures but not the primary moving segments. This technical note proposes an index that quantitatively characterizes different dynamic postural strategies employed during sagittal plane lifting. Dynamic lifting strategies are modeled in the velocity domain as different schemes of partitioning postural changes between the torso and leg segments. The index consists of two parameters, assigned to two leg segments, quantifying their contributions relative to the torso. Given a measured lifting movement, its index parameters values, ranging from 0.1 to 10, are estimated through an enumeration search process with the objective of minimizing the fitting error. The use of this index is illustrated by applying it to 24 lifting movements performed by six subjects assuming either a back-lift or a leg lift strategy. Results indicate that a lifting strategy, in terms of whether the leg or the back is generally the prime mover, can be differentiated and visualized using this simple two-parameter index. In addition, indistinct intermediate strategies are also discerned, as the involvement of each segment in a lifting movement is quantified. The index is however limited in that it does not accommodate arm motion contributions to a lift nor possible time-dependent strategic changes during a lift. Potential future applications include time efficient movement prediction and simulation for computerized biomechanical or ergonomic analysis. PMID- 10808002 TI - Algorithms to determine event timing during normal walking using kinematic data. AB - Algorithms to predict heelstrike and toeoff times during normal walking using only kinematic data are presented. The accuracy of these methods was compared with the results obtained using synchronized force platform recordings of two subjects walking at a variety of speeds for a total of 12 trials. Using a 60Hz data collection system, the absolute value errors (AVE) in predicting heelstrike averaged 4.7ms, while the AVE in predicting toeoff times averaged 5.6ms. True average errors (negative for an early prediction) were +1.2ms for both heelstrike and toeoff, indicating that no systematic errors occurred. It was concluded that the proposed algorithms provide an easy and reliable method of determining event times during walking when kinematic data are collected, with a considerable improvement in resolution over visual inspection of video records, and could be utilized in conjunction with any 2-D or 3-D kinematic data collection system. PMID- 10808003 TI - Giant pelvic retroperitoneal liposarcoma. AB - BACKGROUND: Pelvic retroperitoneal liposarcomas are rare in young women (under 30 years old). We present a case of a giant well-differentiated liposarcoma of the retroperitoneum in a young woman. CASE: A 27-year-old nulligravida presented with rapid abdominal enlargement. Pelvic examination found a huge mass extending from the left adnexa to the epigastric region. At surgery, the pelvic organs were displaced to the right side by a retroperitoneal mass that marginally involved the left fallopian tube. The left ovary, uterus, and right adnexa were not involved and were not removed. No adjuvant irradiation was given. The patient was alive and well 2 years after the operation and was 23 weeks pregnant. CONCLUSION: Gynecologists should be familiar with pelvic retroperitoneal liposarcomas and their treatment. PMID- 10808004 TI - Uterine myxoid leiomyosarcoma. AB - BACKGROUND: Uterine myxoid leiomyosarcoma is rare, has a poor prognosis, and must be distinguished from a uterine myoma with myxomatous change. CASE: A 56-year-old woman with a history of epigastric pain and generalized abdominal swelling had a peritoneal cavity filled with tumor that originated from the posterior uterine wall. Chemotherapy and two operations for recurrence could not prevent fatal metastases to the vascular system. Histopathology showed a typical myxoid leiomyosarcoma. CONCLUSION: Although the patient's prognosis was poor, no mitoses were observed, and the nuclear abnormalities were not sufficient to diagnose the tumor as malignant at the initial surgery. Whenever a uterine myoma with myxomatous change is found, the patient should be monitored carefully. PMID- 10808005 TI - Colpectomy after vaginoplasty in transsexuals. AB - BACKGROUND: Penile amputation, vaginoplasty, and clitoroplasty help male-to female transsexuals accept their bodies and increase psychosocial function. Subsequent colpectomy is unusual. CASES: We report three patients in whom complicated, long-term problems subsequent to vaginoplasty led to total colpectomy in one case of neovaginal overgrowth of condylomata acuminata, and in two cases of colitis in rectosigmoid transplants used for neovaginoplasties. CONCLUSION: Before inversion of penile skin, in cases in which the genital skin has condylomata, the risk of condylomata overgrowth might be anticipated. Rectosigmoid vaginoplasty might result in therapy-resistent colitis, which also could lead to colpectomy. PMID- 10808006 TI - Luteinizing hormone receptor expression in leiomyomatosis peritonealis disseminata. AB - BACKGROUND: Leiomyomatosis peritonealis disseminata has been attributed to estrogen stimulation and is seen only rarely in postmenopausal women. In such cases, pathogenesis is uncertain. CASE: Leiomyomatosis peritonealis disseminata tumors were resected from a postmenopausal woman. She was receiving tamoxifen therapy for breast cancer and had bilateral ovarian Brenner tumors. Estrogen and progesterone receptors were detected. Immunohistochemical analysis indicated that LH receptors were present. CONCLUSION: Luteinizing hormone receptors were identified in leiomyomatosis peritonealis disseminata in one woman. Levels of FSH and LH increase after menopause, and immunohistochemical analysis showed the presence of LH receptors, so gonadotropin rather than estrogen stimulation might have contributed to development of leiomyomatosis peritonealis disseminata in this uncommon case. PMID- 10808007 TI - Reversed diastolic umbilical artery flow in the first trimester associated with chromosomal fetal abnormalities or cardiac defects. AB - BACKGROUND: Six cases have been reported of reversed end-diastolic umbilical artery (UA) blood flow at 10-14 weeks' gestation. Four were associated with chromosomal anomalies, and one showed congenital heart disease. CASES: This report describes two cases of reversed end-diastolic UA flow at 10 and 12 weeks' gestation with increased nuchal translucency in which cytogenetic analysis by chorionic villus sampling showed triploidy (69,XXX) and trisomy 9, respectively. In second case, the fetus had a ventricular septal defect. CONCLUSION: Reversed end-diastolic UA flow in the first trimester might be an important sign of chromosomal abnormalities in early pregnancy and might be a useful signal for the diagnosis of early cardiac defects. PMID- 10808008 TI - Antenatal ultrasonographic appearance of a cloacal anomaly. AB - BACKGROUND: Cloacal anomalies are rare abnormalities with highly variable forms, which makes them difficult to diagnose antenatally. We present a cloacal anomaly case with interesting antenatal ultrasonographic findings. CASE: A 24-year-old woman, gravida 1, para 0, presented at 23 weeks' gestation with ultrasonographic evidence of fetal ascites. Between 23 and 30 weeks' gestation, serial ultrasounds showed changing features of a cloacal anomaly, consisting of fetal ascites, a cystic pelvic mass, hydronephrosis, and oligohydramnios. CONCLUSION: Serial antenatal ultrasound helped diagnose changing features of a cloacal anomaly in our case, thus allowing directed antenatal counseling and appropriate perinatal treatment. PMID- 10808009 TI - Ultrafast magnetic resonance imaging of cephalopagus conjoined twins. AB - BACKGROUND: Ultrafast magnetic resonance imaging (MRI) has been used successfully in antenatal diagnosis. CASE: We report a case of cephalopagus conjoined twins in which ultrafast MRI clarified how the twins were united, especially the neuroanatomy, and helped with antenatal counseling. CONCLUSION: Ultrafast MRI can provide superior image quality to two-dimensional ultrasonography and should be considered an adjunct to ultrasound for antenatal characterization of some anomalies. PMID- 10808010 TI - Cortical blindness in severe preeclampsia: computed tomography, magnetic resonance imaging, and single-photon-emission computed tomography findings. AB - BACKGROUND: Cortical blindness is a complication of severe preeclampsia, but it is unclear whether it results from cerebral vasospasm and ischemic injury or vasogenic (hydrostatic) edema due to increased capillary permeability. CASE: Reversible cortical blindness in a 33-year-old gravida 2, para 1, with severe postpartum preeclampsia after evacuation of a partial molar pregnancy at 19 weeks' gestation is presented. Initial neuroimaging studies showed hyperperfusion on head single-photon-emission computed tomography scan, which corresponded with lesions found on head computed tomography and magnetic resonance imaging scans. Follow-up neuroimaging studies 2 weeks later, by which time the patient's visual acuity had returned to normal, showed complete resolution of radiologic abnormalities. CONCLUSION: Neuroimaging studies in a woman with severe postpartum preeclampsia complicated by reversible cortical blindness showed that blindness resulted from vasogenic (hydrostatic) cerebral edema and not cerebral vasospasm. PMID- 10808011 TI - B-Lynch suture for postpartum hemorrhage. AB - BACKGROUND: Postpartum hemorrhage is a major contributor to maternal morbidity and mortality. Numerous medical and surgical therapies have been used, but none has been uniformly successful. CASE: Two women with postpartum hemorrhage due to uterine atony after cesarean for twins are presented. Neither responded to medical management. In the first subject, O'Leary uterine artery ligation and utero-ovarian branch ligations were done without benefit. The B-Lynch suture immediately sustained correction of hemorrhage in both subjects. Magnetic resonance imaging and hysterosalpingogram after the first case showed no uterine defects. CONCLUSION: The B-Lynch suture might be a valuable addition to the surgical treatment of postpartum hemorrhage due to uterine atony. PMID- 10808012 TI - Hemophagocytic syndrome in pregnancy. AB - BACKGROUND: Hemophagocytic syndrome is characterized by nonmalignant histiocytes that undergo uncontrolled phagocytosis of normal hematopoietic cells. Clinical severity ranges from complete recovery to rapid deterioration and death. CASE: Thrombocytopenia was discovered upon routine initial prenatal evaluation of a 24 year-old, gravida 2, para 1, at 29 weeks' gestation with a history of necrotizing lymphadenitis. Cytopenia and elevated transaminases developed, followed by hyperpyrexia. The patient delivered and her postpartum course was complicated by coagulopathy, multiorgan failure, and death. Bone marrow biopsy confirmed hemophagocytic syndrome. CONCLUSION: Early diagnosis of hemophagocytic syndrome during pregnancy might be helped by recognizing symptoms and signs, including a history of necrotizing lymphadenitis, and obtaining a bone marrow biopsy. PMID- 10808013 TI - Anaphylaxis after laminaria insertion. PMID- 10808014 TI - Parvovirus infection after laparoscopic hysterectomy using fibrin glue hemostasis. PMID- 10808015 TI - Cat scratch disease presenting as a breast mass. PMID- 10808016 TI - Incisional hernia involving a fallopian tube and mimicking scar endometriosis. PMID- 10808017 TI - Mesh erosion into the bladder after abdominal sacral colpopexy. PMID- 10808018 TI - Acute vulvar vestibulitis occurring during chemotherapy with cryptophycin analogue LY355703. PMID- 10808019 TI - Leydig cell tumor, mature teratoma, and nongestational choriocarcinoma in a single ovary. PMID- 10808020 TI - Autoimmune hepatitis in pregnancy. PMID- 10808021 TI - Successful pregnancy and spinal muscular atrophy. PMID- 10808022 TI - Bartter syndrome in pregnancy. PMID- 10808023 TI - Maternal thyrotoxicosis and fetal nonimmune hydrops. PMID- 10808024 TI - Hypokalemic periodic paralysis in pregnancy after 1-hour glucose screen. PMID- 10808025 TI - Human caudal appendage diagnosed prenatally with ultrasound. PMID- 10808026 TI - Antenatal doppler diagnosis of fetal ovarian torsion. PMID- 10808027 TI - Fetal bladder outlet obstruction caused by ureterocele. PMID- 10808028 TI - External cephalic version after rupture of membranes. PMID- 10808029 TI - Auxiliary liver transplant in acute fatty liver of pregnancy. PMID- 10808030 TI - Dysfibrinogenemia and placental abruption. PMID- 10808031 TI - Late septic necrosis and dehiscence of a cesarean incision. PMID- 10808032 TI - Sexuality after hysterectomy. AB - OBJECTIVE: The availability of alternative therapies for abnormal gynecologic bleeding has prompted a reexamination of the impact of total hysterectomy on a woman's quality of life. This paper examines the evidence about the impact of hysterectomy on one aspect of quality of life, sexuality. DATA SOURCES: A systematic review of the English and German language literature was conducted using the search terms "sexuality," "hysterectomy," "libido," "orgasm," and "outcomes." Additional papers found in the bibliographies of theses papers were reviewed. METHODS OF STUDY SELECTION: Eighteen studies were found, eight prospective and ten retrospective. With a few exceptions, the methodologic quality of the studies was poor, but we opted to review all of the studies given the paucity of data on this important subject. TABULATION, INTEGRATION, AND RESULTS: The studies were evaluated for methodologic quality using a scoring system described in the paper and for their findings concerning the impact of hysterectomy on sexuality. chi(2) and Fischer exact test were used to analyze nominal variables. Outcome measures were usually not validated and most studies did not consider important confounding factors. Most studies in this review showed either no change or an enhancement of sexuality in women who had a hysterectomy. CONCLUSION: The majority of research evaluating the effect of hysterectomy on sexuality was poorly designed. The available evidence shows that quality of life is improved for most women who had hysterectomy and that hysterectomy did not adversely affect sexuality. A number of confounding factors with the potential to have either a positive or negative impact on sexuality, independent of hysterectomy, should be taken into account in future studies. PMID- 10808033 TI - Intrapartum amnioinfusion for meconium-stained fluid: meta-analysis of prospective clinical trials. AB - OBJECTIVE: To evaluate the effectiveness of intrapartum prophylactic amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid. DATA SOURCES: We identified prospective clinical trials of amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid (AF) published in English by using computerized databases, references in published studies, and index reviews. METHODS OF STUDY SELECTION: We analyzed prospective studies of intrapartum amnioinfusion for meconium-stained AF. In every case, group allocation was based exclusively on meconium in AF. Only published studies with clearly documented outcome data were included. The trials were evaluated for meconium below the vocal cords, meconium aspiration syndrome, fetal acidemia, cesarean delivery, and postpartum endometritis. Each trial was evaluated for the quality of its methodology, inclusion and exclusion criteria, adequacy of randomization, amnioinfusion protocols, definition of outcomes, and statistical analyses. TABULATION, INTEGRATION, AND RESULTS: Thirteen studies met inclusion criteria for our systematic review. Odds ratios (ORs) with their 95% confidence intervals (CIs) were calculated for each outcome. Estimates of ORs and risk differences for dichotomous outcomes were calculated using random and fixed effects models. We tested for homogeneity across the studies. We found that intrapartum amnioinfusion significantly reduced the frequency of meconium aspiration syndrome (OR 0.30; 95% CI 0.19, 0. 46), of meconium below the vocal cords, and neonatal acidemia. Subjects allocated to receive amnioinfusion also had a significantly lower overall cesarean rate (OR 0.74, 95% CI 0.59, 0.93) without increased postpartum endometritis. CONCLUSION: Amnioinfusion in cases of meconium-stained fluid significantly improves neonatal outcome, lowers the cesarean delivery rate, and does not increase the postpartum endometritis rate. PMID- 10808034 TI - Amniotic fluid interleukin-6 and preterm delivery: a review. AB - OBJECTIVE: To evaluate the potential role of amniotic fluid (AF) interleukin (IL) 6 as a predictor of preterm delivery and to consider possible explanations for the proportion of women with elevated AF IL-6 who deliver preterm yet lack microbiologically detectable intra-amniotic infection. DATA SOURCES: We searched the English language human literature in MEDLINE, 1966 through September 1999, using the keywords "labor/infant," "premature," "cytokines/interleukin-6," and "AF." We also examined abstracts from the 1999 meetings of the Society for Maternal-Fetal Medicine and the Society for Epidemiologic Research. We identified other studies by reviewing the reference lists of published articles. METHODS OF STUDY SELECTION: The MEDLINE search yielded 55 citations. We focused on studies that reported on the association between AF IL-6 and preterm delivery. TABULATION, INTEGRATION, AND RESULTS: There is consensus in the literature that elevated AF IL-6 is a stronger predictor of preterm delivery than intra-amniotic infection detected by either microbiologic culture or polymerase chain reaction (PCR). Among women with elevated AF IL-6, PCR could detect a higher proportion of intra-amniotic infection than culture. A number of women with elevated AF IL-6 (33-70%) deliver preterm and do not have evidence of intra-amniotic infection by either culture or PCR. Possible explanations for this observation are considered. CONCLUSION: Elevated AF IL-6 is strongly associated with preterm delivery and merits future consideration in clinical settings to predict preterm delivery and guide patient care. Development of improved polymerase chain reaction-based clinical methods to detect intra-amniotic infection is necessary to better understand the relationship between elevated AF IL-6, intra-amniotic infection, and preterm delivery. PMID- 10808035 TI - Trouble sleeping and anxiety/depression in childhood. AB - The purpose of this report was to estimate the association between children's trouble sleeping and anxiety/depression at ages 6 and 11, cross-sectionally and prospectively. Data come from a study of the psychiatric sequelae of low birth weight (LBW: <2500 g). LBW and normal birth weight children were randomly selected from the 1983-1985 newborn lists of an urban and a suburban hospital. Eight hundred and twenty-three children participated at age 6 and, of those, 717 (87.1%) participated at age 11. Achenbach's Child Behavior Checklist (CBCL) and the Teacher Report Form (TRF) were used to obtain ratings of psychiatric problems. The CBCL asked if the child had trouble sleeping during the past 6 months. Children with trouble sleeping had significantly increased odds of anxiety/depression based on mothers' reports (OR=6.9, 95% CI 4.1-11. 4) but not teachers' reports (OR=1.1, 95% CI 0.4-2.7). There was a greater association between sleep and depression at age 11 than at age 6, and among suburban than among urban children. These findings remained when adjusted for birthweight, sex, and mother's history of major depressive disorder. Profile analysis indicated a stronger association of trouble sleeping with anxiety/depression than other psychiatric problems. The association of trouble sleeping at age 6 with incidence of depression at age 11 was not statistically significant (suburban children RR=2.22, 95% CI 0.53-9.23; urban children RR=0.92, 95% CI 0.20-4.18). PMID- 10808036 TI - Seasonal variations in internalizing, externalizing, and substance use disorders in youth. AB - Seasonal variations were assessed in symptoms of internalizing (anxiety and mood), externalizing (attention-deficit/hyperactivity and oppositional defiant disorders), and substance use disorders in youth. This study is based on secondary data analysis of two NIMH-funded epidemiologic-services studies: (a) Alternative Service Use Patterns by Youth with Serious Emotional Disturbance (N=936, ages 9-17); and (b) Methods for the Epidemiology of Child and Adolescent Mental Disorders (N=1285, ages 9-17). Child psychiatric diagnoses were measured by the Diagnostic Interview Schedule for Children. Variables that indicate site of the interview and service system, as well as age, gender, and ethnicity, were used as covariates. Significant annual variations were found in symptom counts of overanxious disorder, obsessive-compulsive disorder, separation anxiety disorder, social phobia, and major depressive disorder, with the estimated nadir in August October. There may be weak seasonal variations in attention-deficit/hyperactivity disorder with estimated nadir in August, oppositional defiant disorder with estimated nadir in August-September, and marijuana use with estimated zenith in August-September. Significant seasonality in alcohol, other substance use, agoraphobia, and panic disorder was not found. There may be an instrument specific bias in estimated nadir. Real nadirs may be up to 3 months prior to the estimated nadirs specified above. Findings suggest that seasonality in symptoms should be considered when assessment instruments of childhood psychiatric disorders are developed, as well as when epidemiological and clinical data are collected and analyzed. PMID- 10808037 TI - Circulating leptin in patients with anorexia nervosa, bulimia nervosa or binge eating disorder: relationship to body weight, eating patterns, psychopathology and endocrine changes. AB - A decreased production of leptin has been reported in women with anorexia nervosa (AN) and has been attributed merely to the patients' reduced body fat mass. The extent to which eating patterns, purging behaviors, psychopathology and endocrine changes may contribute to the genesis of leptin alterations has not been deeply investigated. Therefore, we measured plasma levels of leptin, glucose and other hormones in three groups of eating disorder patients with different body weight (BW), eating patterns and purging behaviors. Sixty-seven women, 21 with AN, 32 with bulimia nervosa (BN), 14 with binge-eating disorder (BED) and 25 healthy females volunteered for the study. We found that circulating leptin was significantly reduced in AN and BN patients, but significantly enhanced in women with BED. In anorexics, plasma glucose was decreased, whereas plasma cortisol was enhanced; blood concentrations of 17beta-estradiol and prolactin (PRL) were reduced in both AN, BN and BED patients. In all subject groups, a strong positive correlation emerged between plasma levels of leptin and the subjects' BW or body mass index, but not between leptin and psychopathological measures, plasma glucose, cortisol, PRL and 17beta-estradiol. Since leptin was reduced in both underweight anorexics and normal weight bulimics, but increased in overweight BED women, who compulsively binge without engaging in compensatory behaviors, we suggest that factors other than BW may play a role in the determination of leptin changes in eating disorders. PMID- 10808038 TI - Binge eating disorder: prevalence and psychopathological features in a clinical sample of obese people in Italy. AB - Binge eating disorder (BED) is a recently conceptualized eating disturbance, and its clinical features and prevalence are still a matter of debate. This study uses interview methodology to estimate the prevalence of BED in Italy in a sample of 66 obese people presenting for treatment, and examines potential related features typical of patients with anorexia and bulimia nervosa. The lifetime and the 6-month prevalences of BED were 18.1 and 12.1%, respectively. Breaking the group down on the basis of the current as well as lifetime presence of BED, we found that the weight and shape primary to self-esteem, and the interpersonal distress related to body image, were associated with lifetime BED (P<0.05). All or-none thinking about food and dieting was typical of BED patients as a whole, either current (P<0.01) or remitted (P<0.05). We discuss two important findings from our study: (a) the key role of self-esteem depending upon weight and shape in discriminating the eating-disordered obese from non-eating-disordered individuals; and (b) the need to explore the whole lifespan when screening for BED, so pointing to the state-trait issue. PMID- 10808039 TI - Increase in first admissions for schizophrenia and other major psychoses in Italy. AB - Despite reports of falling first-admission rates for schizophrenia in some Western countries, methodological problems and bias preclude a definite conclusion about a genuine fall in the incidence of schizophrenia. This study set out to test the hypothesis that first admissions for schizophrenia in Italy have fallen in recent years. All admissions rated as 'first contact' in Italy from 1984 to 1994 for severe mental illnesses to general hospital psychiatric services, as reported in the Italian National Institute for Statistics Health Care Yearbooks, were considered. Data were analyzed as rates per 100000 in the general population, and changes over time in incidence of schizophrenia, paranoia, affective psychoses and drug-induced psychoses (diagnosed according to ICD 9) were recorded. Changes in rates over time, with rates as the dependent variable and years as the independent variable, were the main outcome measure. First-admission rates for schizophrenia and paranoia increased progressively from 1984 to 1994, as did those for affective psychoses, mania and, to a lesser extent, major depression. In the same period, all admissions (both total and rated as 'first-contact') for mental disorders increased. Although linear regression tests for admission rates in most, but not all, cases indicate a significant ascending linear trend, quadratic model results show a significantly better fit than does the simple linear regression model for the majority of data. The change described by the quadratic model suggests an increase in admission rates more marked in the second half than in the first half of the period of the study. First-admission rates for schizophrenia and, to a lesser extent, paranoia seemed to increase concurrent to a decrease in first-admission rates for 'other' non-organic psychoses. Contrary to reports from other Western countries, hospital incidence in Italy for schizophrenia is on the increase, as is that for other severe psychoses. This increase is likely to be a reflection of changes in mental health-care organisation, in treatment and diagnostic patterns, and in cultural attitudes towards mental illness. Radical changes in the true incidence of psychoses, in particular of mood disorders, as described elsewhere, cannot be ruled out as contributing factors. Data bias and limitations preclude a generalisation of results, however. PMID- 10808040 TI - Physiological characteristics of platelet/circulatory serotonin: study on a large human population. AB - The aim of this work was the study of platelet/circulatory serotonin (5 hydroxytryptamine, 5-HT), specifically alternative ways of its measurement and main physiological characteristics. The study was performed on a large human population (N=500) of blood donors of both sexes over the course of a longer time period (17 months). Owing to the heterogeneity in measurement of circulatory serotonin encountered in the literature, three ways of expression were comparatively studied: per unit number of platelets, per unit mass of platelet protein and per unit volume of whole blood. Results demonstrated unimodal distribution of individual frequencies of platelet/circulatory serotonin in the human population with the mean values of 579+/-169 ng 5-HT/10(9) platelets; 332+/ 89.9 ng 5-HT/mg protein and 130+/-42.3 ng 5-HT/ml blood (mean+/-S.D.). A progressive decrease of serotonin level with age (18-65 years) was demonstrated, reaching statistical significance between the extreme age groups. No significant differences in the serotonin level between the sexes were observed. No seasonal oscillations in platelet/circulatory serotonin were found. Platelet serotonin demonstrated intra-individual stability over time. Finally, regarding the methodology of measurement, our results demonstrated a good correlation among the above-mentioned ways of expression of platelet/circulatory serotonin. This indicates the possibility of intercomparison of the literature reports expressing this physiological parameter either as 5-HT concentration in platelets or as 5-HT level in the circulation. PMID- 10808041 TI - The Greek version of the calgary depression scale for schizophrenia. AB - The aim of this study was to evaluate the reliability and validity, as well as the specificity, of the Greek version of the Calgary Depression Scale for Schizophrenia (CDSS). Schizophrenic inpatients consecutively admitted at the Eginition Hospital, University of Athens, were included in the study. Patients were assessed on admission using the CDSS, the Hamilton Depression Rating Scale (HDRS), the Positive and Negative Syndrome Scale (PANSS), the Rating Scale for Extrapyramidal Side Effects (RSESE), the Rating Scale for Drug-Induced Akathisia (RSDIA) and the Abnormal Involuntary Movement Scale (AIMS). The CDSS was found to have a high inter-rater reliability, as well as test-retest reliability or split half reliability. The internal consistency of the CDSS was good (a=0.87). There were positive correlations between the CDSS and the HDRS, or the depression cluster of the PANSS. The mean score on the CDSS showed no significant correlations with that of the PANSS negative subscale (r=0.123); a negative but not significant correlation with that of the PANSS positive subscale (r=-0.036); a weak correlation with that of the PANSS general psychopathology subscale (r=0.218); and no significant correlations with that of the RSESE (r=0.197), the RSDIA (r=0.160) or the AIMS (r=0.031). Our results give further support to the reliability, the validity, and the specificity of the CDSS. PMID- 10808042 TI - Increased growth hormone response to sumatriptan challenge in adult autistic disorders. AB - Serotonergic (5-HT) abnormalities have been documented in autism. To assess sensitivity of the 5-HT1d receptor, growth hormone response to the 5-HT1d receptor agonist sumatriptan was studied in adult autistic patients and matched normal controls. In this study, 11 adult patients with autism or Asperger's disorder were compared with nine matched controls. All subjects were randomized to single dose sumatriptan (6 mg SQ) and placebo challenges, separated by a 1 week interval, and growth hormone was measured before and during the challenges. The results showed a highly significant diagnosisxdrugxtime interaction on repeated measure analysis covaried for baseline. This suggests that autistic patients had significantly greater growth hormone response to sumatriptan than normal controls, independent of placebo effects. Therefore, abnormalities in 5-HT regulation in autism may be related to increased sensitivity of the 5-HT1d inhibitory receptor in autism. PMID- 10808043 TI - Double-blind acute clonazepam vs. placebo in carbon dioxide-induced panic attacks. AB - The inhalation of 35% carbon dioxide has consistently been shown to provoke panic attacks in panic disorder patients. We aim to determine if an acute dose of clonazepam (2 mg) attenuates the panic attacks induced by an inhalation of 35% carbon dioxide in panic disorder. Twenty-two panic disorder patients who had been drug-free for 1 week participated in a carbon dioxide challenge test 1 h after a dose of either 2 mg of clonazepam or placebo with a randomized double-blind method. Also in a double-blind design during the tests the patients inhaled either atmospheric compressed air ('placebo control') or the carbon dioxide mixture. All patients participated in both tests which were done with a 20-min interval. Immediately before and after the inhalation, the anxiety levels and the symptoms of panic were always assessed. In the clonazepam group (n=11) two patients (18.2%) had a mild panic attack and in the placebo group (n=11) nine patients (81.8%) had a moderate to severe panic attack in the CO(2) challenge test. No patient had panic attacks during inhalation of atmospheric compressed air although anticipatory anxiety levels tended to be higher than in the CO(2) tests. After the CO(2) test anxiety levels were significantly greater in the CO(2) group (three-way ANOVA with Geisser-Greenhouse adjustments, F(31.92,1.86)=17.15, d.f.=7, P=0.013). Although a small sample was studied, the findings suggest the efficacy of an acute dose of clonazepam in attenuating panic attacks induced by carbon dioxide inhalation. PMID- 10808044 TI - Patterns of symptom improvement during antidepressant treatment of delusional depression. AB - We assessed the pattern of changes in depressive symptoms in delusional depressed inpatients treated openly with 300 mg/day of fluvoxamine for 6 weeks. We studied 59 inpatients affected by bipolar (n=23) and major depressive (n=36) disorders with psychotic features (DSM-IV) who showed complete responses to fluvoxamine treatment. Responses were evaluated using the Hamilton Rating Scale for Depression (HAMD-21, divided into: Core, Activity, Psychic anxiety, Somatic anxiety and Delusion clusters) administered at baseline and weekly until the 6th week. Random Regression Model (RRM) analysis was used to investigate the longitudinal time course of HAMD clusters. HAMD depressive symptom clusters decreased in a parallel manner from baseline to the end of the 6-week trial. The RRM analysis revealed no significant difference between HAMD clusters and the time course of the total HAMD score during treatment. Our data indicate that there is a simultaneous decrease in depressive symptoms during antidepressant treatment of delusional depressives. PMID- 10808045 TI - Experimental allergic neuritis in the SJL/J mouse: induction of severe and reproducible disease with bovine peripheral nerve myelin and pertussis toxin with or without interleukin-12. AB - We report a reproducible model of experimental allergic neuritis (EAN) with severe clinical signs and consistent pathological features in mice. Pertussis toxin (PT) in the presence or absence of murine recombinant interleukin-12 (mrIL 12) was used as an adjuvant with bovine peripheral nerve myelin (BPNM) to induce clinical EAN in SJL/J mice. After immunization with a combination of BPNM in complete Freund's adjuvant (CFA) and PT, mice developed severe consistent signs of EAN. The additional treatment of immunized mice with mrIL-12 prolonged the course of EAN characterized by earlier clinical signs of the disease and delayed the recovery stage. Mice injected with BPNM and CFA without PT developed mild clinical signs. Histological examination of the caudae equinae and the sciatic nerves taken from mice with clinical signs of EAN during the recovery stage revealed severe demyelination, remyelination and remnants of mononuclear cell infiltration. Moderate to severe EAN can be induced in SJL/J mice by the injection of a combination of BPNM in CFA and PT. This model can provide a better understanding of mechanism of demyelination in infiltrating peripheral neuropathy. PMID- 10808046 TI - IL-1beta immunoreactive neurons in the human hypothalamus: reduced numbers in multiple sclerosis. AB - Corticotropin-releasing hormone (CRH)-containing neurons in the paraventricular nucleus (PVN) in the hypothalamus of multiple sclerosis (MS) patients are hyperactivated. Since interleukin-1 (IL-1)beta is a powerful activator of CRH neurons, its immunohistochemical expression was studied in the postmortem hypothalamus of MS patients (n=11) and matched controls (n=11). Hypothalamic tissue of 10/11 MS patients showed demyelinating lesions that in many cases contained IL-1beta-immunoreactive (ir) macrophages and glial cells. In control subjects IL-1beta-ir was only sporadically found in glial cells. Interestingly, abundant IL-1beta-ir was also present in hypothalamic neurons. Neuronal IL-1beta co-localised with oxytocin and not with vasopressin or CRH. IL-1beta clearly yielded a less intense staining in neurons and numbers of IL-1-ir neurons in the PVN were 4.5-fold reduced in MS. We suggest that IL-1beta produced by activated glial cells in the hypothalamus of MS patients may contribute to the activation of the hypothalamic CRH neurons, while reduced expression of neuronal IL-1beta in MS patients may have consequences for neuroendocrine, behavioural or autonomic functioning. PMID- 10808047 TI - A catalyst of peroxynitrite decomposition inhibits murine experimental autoimmune encephalomyelitis. AB - Peroxynitrite (PN), the product of nitric oxide (NO) reacted with superoxide, is generated at sites of inflammation. Nitrotyrosine (NT), a marker of PN formation, is abundant in lesions of acute experimental autoimmune encephalomyelitis (EAE), and in active multiple sclerosis (MS) plaques. To determine whether PN plays a role in EAE pathogenesis, mice induced to develop EAE were treated with a catalyst specific for the decomposition of PN. Because this catalyst has no effect upon NO, using it allowed differentiation of PN-mediated effects from NO mediated effects. Mice receiving the PN decomposition catalyst displayed less severe clinical disease, and less inflammation and demyelination than control mice. Encephalitogenic T cells could be recovered from catalyst-treated mice, indicating that the PN decomposition catalyst blocked the pathogenic action of PN at the effector stage of EAE, but was not directly toxic to encephalitogenic T cells. PN plays an important role distinct from that of NO in the pathogenesis of EAE, a major model for MS. PMID- 10808048 TI - Respective roles of inflammation and axonal breakdown in the regulation of peripheral nerve hemopexin: an analysis in rats and in C57BL/Wlds mice. AB - We have previously demonstrated that one of the peripheral nerve responses to injury is the overexpression of hemopexin (HPX). Here, we demonstrate that Wallerian degeneration is required for this response, since HPX does not increase in C57BL/Wlds mice, which display a severely impaired Wallerian degeneration. We also show that HPX synthesis is dramatically increased in macrophages during their activation or after IL-6 stimulation. However, IL-6-driven HPX overexpression occurs in vivo and in vitro in the absence of substantial macrophage invasion. We conclude that, after nerve injury, HPX overexpression occurs first in Schwann cells as a result of axotomy and is subsequently regulated by inflammation. Furthermore, our results and those already described suggest that IL-6, synthesized by the various cell types producing HPX, control nerve HPX expression via paracrine and autocrine mechanisms. PMID- 10808049 TI - Identification of novel trkA variants with deletions in leucine-rich motifs of the extracellular domain. AB - The peripheral expression of trkA encoding for NGF receptor was investigated by RNase protection assay. A thymus-specific protected fragment was identified. Using 5' rapid amplification of cDNA ends, three different trkA fragments were characterized. The longer fragment corresponded to the classical trkA L3 transcripts while the two shorter fragments lacked sequences encoding for leucine rich motifs of the extracellular domain of TrkA, similarly to the trkB L1 and L0 variants. RT-PCR analysis of adult rat tissues showed the expression of trkA L1 transcripts in the thymus, testis, lung and kidney but not in the central nervous system. Their combined expression with trkA L3 transcripts suggests that specific peripheral TrkA oligomers may modulate NGF binding and function in non-neuronal cells. PMID- 10808050 TI - Antidepressant drug administration modifies the interactive relationship between alpha(2)-adrenergic sensitivity and levels of TNF in the rat brain. AB - A reciprocally permissive interaction occurs between cellular responses elicited by the pleiotropic cytokine tumor necrosis factor-alpha (TNF) and alpha(2) adrenergic receptor activation, such that each may adapt in response to modifications in the other's effects. Changes in presynaptic adrenergic sensitivity as well as neuronal sensitivity to TNF have been implicated in the mechanism of action of antidepressant drugs. The present study examines the influence of alpha(2)-adrenergic receptor activation on levels of TNF in regions of the brain associated with adrenergic function and the expression of mood. Additionally, the role of TNF as a neuromodulator is demonstrated by in vivo microinfusion of rrTNF proximal to the hippocampus. Administration to rats of an alpha(2)-adrenergic receptor agonist (clonidine) decreases levels of TNF in homogenates of rat locus coeruleus and hippocampus within 7.5 min. Chronic (14 days) administration of the antidepressant drugs desipramine or zimelidine transforms alpha(2)-adrenergic receptor-dependent decreases in TNF levels to increases in levels of TNF in the locus coeruleus. This transformation to an increase in total levels of TNF also occurs, although transiently, in the hippocampus following acute (1 day) antidepressant drug administration. The effect of TNF on presynaptic alpha(2)-adrenergic sensitivity was also investigated. Field stimulation of hippocampal slices from rats microinfused with rrTNF proximal to the hippocampus for 14 days demonstrates a decrease in fractional release of [3H]NE and an increase in alpha(2)-adrenergic autoreceptor sensitivity. These data demonstrate a mutual dependence between alpha(2) adrenergic receptor activation and levels of TNF in the central nervous system that would culminate in an increase in neurotransmitter release following antidepressant drug administration. PMID- 10808051 TI - Th2-dependent airway eosinophilia is regulated by preproenkephalin. AB - We have used preproenkephalin (PPNK)-deficient mice to study the role of PPNK in the development of airway inflammation. Airway eosinophilia was established by either sensitization followed by airway challenge with OVA or by infection with Nippostrongylus brasiliensis. Both models induce a strong Th2 immune response, characterized by an IL-5-dependent airway eosinophilia. We observed that although the accumulation of lymphocytes in the airways of PPNK-deficient mice was similar to that induced in control mice, IL-5 production and eosinophil infiltration were reduced. We conclude from this work that PPNK has a role in enhancing Th2 cell function and that this molecule may be an important target in asthma immunotherapy. PMID- 10808052 TI - The fate of adoptively transferred quiescent encephalitogenic T cells in normal and antigen-tolerized mice. AB - Adoptive transfer of quiescent encephalitogenic T cells to normal syngeneic recipients was without clinical effect. RT-PCR was used to assess localization of an adoptively transferred quiescent encephalitogenic T cell clone in normal and antigen-unresponsive mice prior to or after challenge with neuroantigen/CFA. The T cell clone was not detectable in lymphoid tissues prior to challenge with neuroantigen; however, following challenge, the clone was found in the spleen, lymph nodes and spinal cord of both normal and antigen-tolerized mice. The latter animals remained clinically normal. Non-activated encephalitogenic T cells transferred to wild-type recipients pretreated i.p. with neuroantigen/IFA were rendered unresponsive. Transfer of the same T cells to alpha/beta T cell deficient mice pretreated with neuroantigen/IFA resulted in spontaneous disease indicating that an intact alpha/beta T cell system was required for development of the unresponsive state. PMID- 10808053 TI - Interleukin-12 enhances clinical experimental autoimmune myasthenia gravis in susceptible but not resistant mice. AB - Experimental autoimmune myasthenia gravis (EAMG) is induced by antibodies against the nicotinic acetylcholine receptor (AChR). Studies indicate a role for interferon-gamma (IFN-gamma) in EAMG. We examined the effect of IL-12, a major inducer of IFN-gamma production, on EAMG in C57BL/6 mice. Five doses of IL-12 accelerated and enhanced clinical disease in AChR-immunized mice. Control B6 mice, IFN-gamma gene-knockout mice, and EAMG-resistant bm12 mice showed no enhancement of disease. Shifting to a Th1-type antibody isotype distribution was insufficient to cause disease. Other factors, such as direct effects of Th1 cytokines on muscle tissue, may be involved in EAMG susceptibility. PMID- 10808054 TI - Experimental autoimmune myositis in the lewis rat: lack of spontaneous T-cell apoptosis and therapeutic response to glucocorticosteroid application. AB - Recently, it has been shown that inflammatory T-cells in human idiopathic myositis only very rarely undergo spontaneous apoptosis. The animal model of experimental autoimmune myositis (EAM) in the Lewis rat was chosen to investigate whether similar findings hold true in rat muscle and if glucocorticosteroids act by induction of T-cell apoptosis in inflammatory lesions. The rate of spontaneous T-cell apoptosis in rat EAM was low, even in muscle specimens with extensive inflammation. After intravenous glucocorticosteroid pulse therapy we found a dramatic increase in the rate of apoptotic T-cells in the inflamed muscles. Up to 50% of these apoptotic T-cells were CD8 positive apoptotic T-cells. T-cell apoptosis was significantly lower in similarly inflamed muscle specimens of the control group. We suggest that glucocorticosteroids induce apoptosis of endomysial T-cells in human idiopathic polymyositis. Glucocorticosteroid-induced apoptosis may be a candidate mechanism in the termination of inflammatory activity. PMID- 10808055 TI - Vasoactive intestinal peptide (VIP) inhibits TGF-beta1 production in murine macrophages. AB - Vasoactive intestinal peptide (VIP) and the structurally related neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP), produced and/or released in the lymphoid microenvironment act primarily as macrophage- and T cell deactivating agents. In the present study we investigate the effect of VIP and PACAP on the production of TGF-beta1 in the macrophage cell line Raw 264.7 and in peritoneal macrophages. The two neuropeptides do not affect the baseline TGF beta1 production by unstimulated macrophages, but reduce dramatically TGF-beta1 production by LPS-stimulated macrophages. The effects are mediated through the specific receptors VPAC1, VPAC2, and PAC1. The effect of VIP is mediated primarily through the cAMP pathway, whereas PACAP activates both the cAMP and the protein kinase C pathway. VIP reduces the TGF-beta1 steady-state mRNA levels in both peritoneal macrophages and Raw 264.7 cells treated with LPS. A similar effect is observed upon the in vivo administration of VIP. This report adds VIP and PACAP to the only other neuropeptide, substance P, known to regulate TGF beta1 production in immune cells. PMID- 10808056 TI - RANTES production and expression is reduced in relapsing-remitting multiple sclerosis patients treated with interferon-beta-1b. AB - RANTES (regulated upon activation, normal T-cell expressed and secreted), a CC chemokine, appears to play a role in the pathogenesis of relapsing-remitting multiple sclerosis (RR-MS), enhancing the inflammatory response within the nervous system. We have demonstrated that RANTES production is increased in RR-MS compared to controls. Interferon-beta-1b (IFN-beta-1b) treatment reduces RANTES production in sera and peripheral blood adherent mononuclear cell (PBAM) supernatants both in relapse and remission. IFN-beta-1b also reduces RANTES expression in PBAM. Our results suggest that RANTES modulation might represent one of the mechanisms of action of IFN-beta-1b in RR-MS. PMID- 10808057 TI - Diurnal blood pressure variations in incipient and end stage diabetic renal disease. AB - Our aim was to compare the diurnal blood pressure patterns of people with Type 1 diabetes on continuous ambulatory peritoneal dialysis (CAPD, n=9) or haemodialysis (n=10) to diabetic patients with normo-albuminuria (n=12) or micro albuminuria (n=15). Blood pressure was measured with an ABPM02 Meditech oscillometric blood pressure monitor. The micro-albuminuric group had significantly higher nocturnal diastolic and mean arterial pressures than the normo-albuminuric group. CAPD and haemodialysis patients had significantly higher day time, nocturnal mean systolic, diastolic and mean arterial blood pressures. Micro-albuminuric and end-stage renal failure patients displayed a loss of the physiological drop of systolic blood pressure, which was only significant in the normo-albuminuric group. Nocturnal drop of blood pressure characterised by diurnal indices were 7.4% in the CAPD, 8.8% in the haemodialysis, 10.0% in the micro-albuminuric and 16.5% in the normo-albuminuric group. These results suggest, that pathological circadian blood pressure variation is common in diabetic patients on dialysis, and ambulatory blood pressure monitoring can be a useful tool both in its the detection and its adequate treatment. PMID- 10808058 TI - Insulin antibody response to bovine insulin therapy: functional significance among insulin requiring young diabetics in India. AB - The majority of young diabetics in India prefer to use low-cost bovine insulin for economic reasons. Therefore, the question of insulin antibody response to bovine insulin and its functional significance is still relevant in the Indian context. We assessed insulin antibody response in 52 young diabetics (type 1, n=25, malnutrition modulated form of diabetes, n=19 and fibrocalculous pancreatopathy (FCP) n=8) on bovine insulin therapy (mean duration 3.0+/-2.1 years) using an internationally standardised in-house radioligand assay. The functional significance of insulin antibody was assessed by calculating their affinity constant, maximum binding capacity and total insulin binding power by Scatchard analysis (type 1, n=14, malnutrition modulated form of diabetes, n=11). All the patients treated with bovine insulin showed high titers of insulin antibodies with S.D. score ranging from 5.1 to 42.0. No significant difference was observed in the mean S.D. score of insulin antibodies in the three diabetic groups. The mean daily insulin dose, maximum insulin binding capacity and total insulin binding power were significantly higher in type 1 when compared to the malnutrition modulated form of diabetes (36+/-8 vs. 26+/-11 IU/day, P<0.05; 9. 7+/-7.8 vs. 4.0+/-3.9 nmol/l, P=0.03 and 59+/-29 vs. 29+/-43, P=0.01, respectively). Insulin antibodies S.D. score and its affinity did not show significant relationship with daily insulin dose and glycemic control (HbAl) at admission. Only 24+/-7% variations in daily insulin requirement were accounted for by total insulin binding power. There was a significant inverse relationship between insulin antibody S.D. score and duration of insulin therapy (r=-0. 4172, P<0.0004). To conclude, insulin antibody response following bovine insulin therapy is not different among type 1, malnutrition modulated form of diabetes and FCP diabetes. The insulin antibody response to bovine insulin therapy does not contribute significantly to increase in daily insulin requirement in bovine insulin treated insulin requiring young diabetics. PMID- 10808059 TI - A type 2 diabetic patient with liver dysfunction due to human insulin. AB - A 45-year-old man had been complaining of thirst and polydypsia for the last 3 months and was diagnosed as having type 2 diabetes mellitus because his fasting blood glucose showed 221 mg/dl with positive urinary ketone. He was hospitalized to a private hospital and Penfil 30R was started. However, serum gamma-GTP and aminotransferases began to elevate after insulin treatment and exceeded 1000 IU/l. Insulin was discontinued and serum gamma-GTP and aminotransferases returned close to the normal range. Since his glycemic control became poor again, Penfil 30R was restarted and serum gamma-GTP and aminotransferases elevated again. Therefore, insulin was discontinued and the patient was referred to the Third Department of Internal Medicine, Yamanashi Medical University Hospital because of liver dysfunction. His plasma glucose decreased by diet therapy, and improved further by the administration of glibenclamide. After obtaining informed consent, Humalin R was challenged. Seven days after insulin injection, serum aminotransferases began to elevate again. Lymphocyte stimulation test was negative against three preparations (Penfil R, Penfil N and Humalin R). The present case suggests that human insulin itself can cause liver dysfunction and we need to pay more attention to liver function tests when we start insulin treatment. PMID- 10808060 TI - Importance of hypercoagulability over hyperglycemia for vascular complication in type 2 diabetes. AB - To critically evaluate the relative importance of coagulation abnormalities over other clinical variables for micro- and macrovascular diabetic complications, prothrombin fragment (F1+2), thrombin-antithrombin III complex (TAT), fibrin degradation product d-dimer, and alpha2 plasmin inhibitor complex were determined in 101 stable, relatively well controlled patients with Type 2 diabetes (the mean HbA1c, age and duration of diabetes, 7.1%, 61 and 7.5 years, respectively). First, incidence and severity of diabetic micro- and macroangiopathies were progressively increased with the severity of coagulation abnormalities. Next, correlation of the four values with the presence of micro- and macrovascular complications, respectively, was analyzed by the multiple logistic regression analysis, with the inclusion of other variables such as age, duration of diabetes, HbA1c, blood pressure, and urinary albumin excretion. With the presence of microangiopathies, F1+2 and systolic blood pressure were significantly related, with the relationship being very strong for the former (P=0.003) and weak for the latter (P=0.035). On the other hand, with the presence of macroangiopathies, F1+2 (P=0.003), TAT (P=0.002), duration of diabetes (P=0.015), and age (P=0.013) were related. Other clinical variables were not significantly related with the presence of complications. Coagulation and fibrinolytic abnormalities are stronger determinants of the presence of diabetic vascular complications than other clinical variables including the degree of glycemia, in stable, relatively well controlled patients with Type 2 diabetes. PMID- 10808061 TI - GAD autoantibodies in a selection-free population of insulin-treated diabetic patients: indicator of a high prevalence of LADA? AB - Up to the present only few data have been available concerning the prevalence of diabetes-specific autoantibodies (anti-GAD, ICA, IAA, IA-2) in unselected populations, in particular in type 2 diabetic patients. Hence, the aim of the present study was to determine the prevalence of anti-GAD in a selection-free population of insulin-treated diabetic patients. Accordingly, 90% of all the insulin-treated diabetic patients (type 1, n=127, type 2, n=117) aged 16-60 years and living in the city of Jena (100242 inhabitants) were examined. In order to test sera for anti-GAD, serum samples were taken in 75% of type 1 (n=95) and in 80% of insulin-treated type 2 diabetic (n=94) patients. RESULTS: In the group of type 1 diabetic patients 55% of the patients tested were positive for anti-GAD. But, interestingly, in the type 2 group, a total of 21% patients were positive. With respect to this high percentage of anti-GAD positive type 2 diabetic patients it must be suggested that the frequency of patients with latent autoimmune diabetes mellitus in adults (LADA) was underestimated in the past. PMID- 10808062 TI - The role of obesity in the association of cardiovascular risk factors and glucose intolerance in small Japanese and North American communities. AB - To investigate whether the influence of obesity on cardiovascular risk factors and glucose intolerance differs between Japan and the US, we conducted cross sectional surveys in those with elevated plasma glucose in Tanushimaru, Japan, and a stratified random population sample, in Wadena, MN. Subjects numbered 204 in Tanushimaru and 334 in Wadena. Body mass index (BMI), blood pressure, blood lipids, fasting plasma levels of glucose, glycosylated hemoglobin A(1c,) insulin, and free fatty acids were assessed. Overweight was defined as BMI of 27.5-29.9 kg/m(2) and obesity as BMI> or =30 kg/m(2). Gradual increases in risk factors were seen as BMI increased in both studies. Most risk factors were associated with glucose intolerance, except for BMI in Tanushimaru. In Wadena, glucose intolerance increased sharply among the obese. Adjustment for BMI attenuated the associations of cardiovascular risk factors with glucose intolerance in Wadena, but not in Tanushimaru. Obesity has an exaggerated influence on risk factors, compared with being overweight. The associations of glucose intolerance with cardiovascular risk factors are more affected by adjustment for BMI in Wadena than in Tanushimaru, not because of a different influence of body weight on risk factors between the two cities, but because obesity is rare in Japan. PMID- 10808063 TI - Association between birth weight and insulin sensitivity in healthy young men in Korea: role of visceral adiposity. AB - Recent studies have demonstrated decreased insulin sensitivity in individuals with low birth weight. This study was performed to examine whether abdominal obesity is a link between insulin resistance and low birth weight. We studied the relationships between birth weight and insulin secretion, insulin sensitivity, and various anthropometric indices including visceral fat area in 22 healthy young Korean adults. Birth weight correlated significantly with diastolic blood pressure (r=-0.47, P<0.05) and insulin sensitivity index (S(I)) measured by a frequently sampled intravenous glucose tolerance test (FSIGT) (r=0.54, P<0.05), but not with insulin secretory indices such as acute insulin responses during FSIGT (r=-0.35, NS) or hyperglycemic clamp (r=0.17, NS) and submaximum insulin response during hyperglycemic clamp (r=0.10, NS). S(I) correlated significantly with abdominal obesity measurements such as waist circumference (r=-0.48, P<0.05), waist-to-hip ratio (r=-0.53, P<0.05) and visceral fat area (r=-0.58, P<0.01). However, we could not find significant correlation between birth weight and any of the abdominal obesity measurements (r=-0.35 for waist-to-hip ratio, r= 0.22 for visceral fat area, and r=-0.24 for visceral-to-subcutaneous fat ratio; NS for all). The present data confirm that low birth weight is associated with insulin resistance in adult life. However, our data suggest that the association between low birth weight and insulin resistance is not mediated by abdominal obesity. PMID- 10808064 TI - Lack of association of the Ala45Thr variant in the BETA2/NEUROD1 with type 1 diabetes in Japanese. AB - To evaluate the role of the Ala45Thr variant of BETA2/NEUROD1 in the development of type 1 or type 2 diabetes, we studied a Japanese population consisting of 383 control subjects, 234 type 1 diabetes patients and 160 type 2 diabetes patients. Both genotypewise and allelewise, there was no significant association of the variant with type 1 diabetes or type 2 diabetes in Japanese. Also, there were no significant differences in clinical characteristics with and without the variant. Our present results do not support a recent report which described an association of the Ala45Thr variant with type 1 diabetes in Japanese. PMID- 10808065 TI - Clinical efficacy of insulin-like growth factor-1 in a patient with autoantibodies to insulin receptors: a case report. AB - The type B insulin-resistance syndrome is characterized by the presence of anti insulin receptor antibodies that cause severe insulin resistance. Treatments including steroids, cyclophosphamide, plasmapheresis, or insulin-like growth factor-1 (IGF-1) are chosen according to severity of insulin resistance. We describe a patient with type B insulin resistance syndrome who was treated successfully with human recombinant (hr) IGF-1, although this treatment provoked a severe allergic reaction. An elderly man with impaired glucose tolerance and unpredictable hypoglycemic episodes which were gradually worsening increased in hemoglobin (Hb)A1c concentration from 6.5 to 13.4%. His fasting and postprandial hyperglycemia were associated with severe hyperinsulinemia. The patient was diagnosed with type B insulin-resistance syndrome by the presence of anti-insulin receptor antibodies. Double-filtration plasmapheresis, plasma exchange, and immunosuppressive therapy with cyclophosphamide and cyclosporin all failed to suppress anti-insulin receptor antibodies more than transiently. When we attempted the treatment by daily administration of hrIGF-1, fasting and postprandial plasma glucose concentrations became normal and HbA1c levels decreased to 7.1% over 2 months, until on one occasion administration resulted in anaphylaxis. After the patient became stable, desensitization therapy was performed successfully, and hrIGF-1 could be administered again with the plasma glucose returning. We concluded that IGF-1 therapy was an effective treatment choice for type B insulin-resistance syndrome in cases whose plasma exchange and immunosuppressive therapy have failed. PMID- 10808066 TI - Increasing anther culture efficiency in rice (Oryza sativa L.) using anthers from ratooned plants. AB - Anther culture response for the variety Taipei-309 was compared using anthers of ratooned and non-ratooned plants. Anthers were plated onto N6 liquid media with 0, 2, 5, and 10 mg l(-1) ABA, respectively. After 5 days in culture, the media were removed and N6 medium without ABA was added. A higher number of pre-mitotic P-pollen grains were observed in anthers from ratooned plants. A significant synergistic effect was found between anthers from ratooned plants/ABA concentrations and the increase in embryogenic-like structures (ELS) induction and green plant regeneration efficiency. The highest frequency of anthers producing ELS was found in N6 medium (6% Sucrose and 2 mg l(-1) 2,4-D) with 5 and 10 mg l(-1) ABA for anthers derived from ratooned plants. Mean green plant regeneration efficiencies were raised from 24.2 to 42% (non-ratooned), and from 30 to 70%, (ratooned donor plants) using a combination of 10 mg l(-1) ABA in induction media, and MS with 2 mg l(-1) BAP, 1 mg l(-1) NAA, 2 mg l(-1) kinetin and 0.45% agarose. The highest number of green shoots per ELS were obtained from ratooned-derived anthers (up to 82). The use of ratooned plants in anther culture, its importance and relative advantages are also discussed. PMID- 10808067 TI - Plant regeneration from callus and suspension cultures of Valeriana edulis ssp. procera via simultaneous organogenesis and somatic embryogenesis. AB - Plants of the genus Valeriana produce irioid esters of biotechnological interest, known as valepotriates. The regeneration of Valeriana edulis ssp. procera via indirect organogenesis and somatic embryogenesis from leaf-derived callus and suspension cultures, is presented. For induction of callus, leaves were cultured on semisolid MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin. Embryogenic and organogenic callus development was observed after 16 weeks. Calli were transferred to semisolid and liquid Murashige and Skoog (MS) medium supplemented with kinetin and 1-naphthaleneacetic acid (NAA). Appearance of shoot and somatic embryo occurred 4 weeks later. Shoots were rooted and somatic embryos were germinated and were transferred to the greenhouse. A better organogenic and embryogenic response was observed from suspension cultures. Histological observations of morphogenic callus revealed that both somatic embryos and shoots arose from the same type of callus. PMID- 10808068 TI - Rose MADS-box genes 'MASAKO C1 and D1' homologous to class C floral identity genes. AB - We isolated AGAMOUS-like cDNA clones called MASAKO C1-C6 (C4-C6 are 3' or 5' partial cDNA clones) and MASAKO D1 from wild rose (Rosa rugosa Thunb. ex Murray). We found that MASAKO C1 was a homologue of AGAMOUS on the basis of sequence similarity, conservation of intron positions, and stamen- and carpel-specific expression within reproductive organs. MASAKO C1-C6 seem to have arisen via alternative splicing, and a possible function of one of the different mRNAs 'MASAKO C6' is discussed in view of similarities between it and the ag-4 variant 2 within Arabidopsis thaliana. MASAKO D1 was also thought to be a homologue of AGAMOUS based on similarity of their sequences, and on the tissue specific expression pattern with GAG2, which is an AGAMOUS homologue in ginseng. Two types of AGAMOUS homologue genes are possibly present in wild rose, as has been observed in tobacco, petunia, cucumber, and maize. PMID- 10808069 TI - Photosynthetic CO(2) assimilation, chlorophyll fluorescence and photoinhibition as affected by nitrogen deficiency in maize plants. AB - Effects of nitrogen deficiency on photosynthetic CO(2) assimilation, PSII photochemistry and photoinhibition were investigated in maize plants grown under natural illumination. Nitrogen-deficient plants had a significantly smaller CO(2) assimilatory capacity, but they showed little changes in the maximal efficiency of PSII photochemistry, the rate of Q(A) reduction and the heterogeneity of PSII reaction centers, suggesting that nitrogen deficiency had little effects on PSII primary photochemistry and photoinhibition even under natural illumination. However, modifications in PSII photochemistry under the steady state of photosynthesis induced by nitrogen deficiency were observed. This is reflected in decreases in the quantum yield of PSII electron transport, the efficiency of excitation energy capture by open PSII reaction centers, and the photochemical quenching coefficient and an increase in the non-photochemical quenching coefficient. These results suggest that modifications of PSII photochemistry under the steady state of photosynthesis may be a mechanism to downregulate photosynthetic electron transport so that production of ATP and NADPH would be in equilibrium with the decreased demand in the Calvin cycle in nitrogen-deficient plants. On the other hand, the nitrogen-deficient plants still exhibited increased susceptibility to photoinhibition when exposed to very high irradiance, although nitrogen deficiency induced no photoinhibition under natural illumination. Our results suggest that such increased susceptibility to photoinhibition was associated with the increased accumulation of inactivated PSII reaction centers, the decreased capacity of non-photochemical quenching, and the increased fraction of the reduction state of Q(A). PMID- 10808070 TI - Identification of an isoform of ornithine carbamoyltransferase that can effectively utilize canaline as a substrate from the leaves of Canavalia lineata. AB - An isoform of ornithine carbamoyltransferase that can utilize effectively canaline as a substrate (C-OCT) was identified from the leaves of Canavalia lineata (Thunb.) DC. The molecular mass of native C-OCT was 109 kDa by Sephacryl S-200 gel filtration and that of the subunit was 36 kDa by SDS-PAGE and immunoblotting using the antibody against kidney bean ornithine carbamoyltransferase (OCT). C-OCT has pH optimum at 8.0 for canaline-dependent OCT activity and Michaelis constants of 9.6 mM for canaline and 0.24 mM for carbamoyl phosphate. To some extent, the C-OCT also showed ornithine-dependent OCT activity and a pH optimum of 8.5; Michaelis constants for ornithine and carbamoyl phosphate were 0.21 and 0.086 mM, respectively. The enzyme exhibit V(max) for canaline-dependent activity 14-fold higher than that for ornithine dependent activity and the ratio of canaline-dependent activity to ornithine dependent activity was 66-fold higher than that of OCT of the same plant. It is likely that this enzyme plays an important role in the canavanine synthesis from the canavanine-containing plants. PMID- 10808071 TI - Scopoletin uptake from culture medium and accumulation in the vacuoles after conversion to scopolin in 2,4-D-treated tobacco cells. AB - Tobacco (Nicotiana tabacum L. Bright Yellow) T-13 cell line has the ability to produce scopoletin endogenously and release some of it into the culture medium. We investigated the mechanism of scopoletin uptake following treatment of a tobacco culture with 2,4-dichlorophenoxyacetic acid (2,4-D). Addition of [14C] labeled scopoletin showed that scopoletin was taken up by 2,4-D-treated cells and converted to scopolin, a 7-O-glucoside of scopoletin. This uptake of scopoletin began 6 h after 2,4-D addition to the cells. Experiments using several inhibitors showed that this uptake was energy-dependent. The phenomenon of 2,4-D-stimulated uptake was observed only for 7-hydroxycoumarins, such as scopoletin, umbelliferone and esculetin. To further investigate the site for scopoletin accumulation, we separated the vacuoles from T-13 cells and quantified the coumarin contents in this fraction. Most of the scopoletin in the vacuoles was present as glucoconjugate, scopolin. Moreover, glucosylation activity was absent from isolated vacuoles and, therefore, is likely to be located in the cytosol. Therefore, we can state that 2,4-D treatment of tobacco cells stimulated scopoletin uptake. The scopoletin was converted into scopolin in the cytoplasm, and then transferred into the vacuoles. PMID- 10808072 TI - Boron and calcium distribution in nitrogen-fixing pea plants. AB - In a glasshouse experiment, plants of Pisum sativum L. cv. Argona were grown hydroponically with different B and Ca levels, in order to elucidate a specific role for B and Ca on the N(2) fixation in this temperate legume. The treatments were as follows: control (9.3 uM B and 2 mM Ca), -B (without B and 2 mM Ca), B+Ca (without B and 3.6 mM Ca), +Ca (9.3 uM B and 3.6 mM Ca), -Ca (9.3 uM B and 0.4 mM Ca) and -Ca+B (46.5 uM B and 0.4 mM Ca). The supply of -Ca and +Ca did not affect nitrogenase activity, but the weight of old shoots and total N content increased with the Ca treatment. No symptoms of B deficiency were detected in the plants of the -B and -B+Ca treatments, apart from weight reduction in young shoots and lower nitrogenase activity. The B concentration decreased in young shoots and roots of plants grown without B (-B), but there was a sharper decrease in the roots of -B+Ca plants and the levels of B in the young shoots were similar to the control levels. The B concentration in -Ca plants was reduced in the old shoot and in the root, while plant weight and N content increased in -Ca+B plants. The cell wall and total B concentrations in the nodules were 4-fold compared with those of the roots. By contrast, the Ca root wall was 2.5 times higher than the nodule levels although total pectin was higher in the nodule than in the root. Finally, the results obtained showed that a high supply of Ca could induce B mobilisation from root to shoot. On the other hand, the high B requirement found in pea plant nodules may contribute to explain the low nitrogenase activity detected under -B conditions. PMID- 10808073 TI - The seed-specific transactivator, ABI3, induces oleosin gene expression. AB - A microspore-derived cell suspension culture of Brassica napus was used as a host for expression studies involving seed oleosin genes. The suspension culture was previously shown to display biochemistry and gene expression typical of zygotic embryos. Using a biolistic, transient expression approach we demonstrate that the seed-specific activator ABI3 promotes oleosin gene expression in these cultures. Co-bombardment of an oleosin promoter-GUS fusion and a full-length ABI3 gene from Arabidopsis resulted in four to six-fold enhancement of GUS expression. Deletion analysis was performed to identify which oleosin upstream sequences were required for ABI3 regulation. These studies found that a truncated oleosin promoter containing 160 bp of 5' regulatory sequence was sufficient to confer ABI3 responsiveness. Mutation of a canonical abscisic acid response element within this 160 bp region had a dramatic effect on basal expression, reducing levels to 25% of control. However, this mutation had no significant effect on ABI3 transactivation, indicating that the reduction in basal oleosin expression was distinct from the ABI3 response. These results also suggest that ABI3-mediated transactivation occurs through either a less-conserved ABRE element or other abscisic acid-independent sequences within the minimal promoter. Together, these data provide the first direct evidence that ABI3 mediates oleosin transactivation. PMID- 10808074 TI - Characterization of the binding of alpha-L-Fuc (1-->2)-beta-D-Gal (1-->), a xyloglucan signal, in blackberry protoplasts. AB - Previous work showed that the fucose-->galactose moiety of the xyloglucan nonasaccharide XXFG is responsable for its biological activity. We used this side chain of XXFG (alpha-L-Fuc (1-->2)-beta-D-Gal (1-->)) in ligand-binding experiments to demonstrate its role as a signal molecule in plant cells. Proteins solubilized from plasma membrane enriched fractions isolated from Rubus fruticosus protoplasts were tested for their ability to bind the side chain of XXFG, using a digoxigenin- or biotin-conjugated neoglycoprotein specific for 2' fucosyl-lactose in blots and k-ELISA tests. A putative receptor for the signaling sugar was identified, and the ligand specificity is reported. The role of structural elements important for biological activities was investigated using compounds structurally related to xyloglucan, and a variety of phytohormones such as 2,4-D. PMID- 10808075 TI - Abscisic acid induced somatic embryogenesis in immature embryo explants of coconut (Cocos nucifera L.). AB - Somatic embryogenesis in coconut (Cocos nucifera L.) is generally induced by gradual reduction of auxin concentration in the culture medium and incorporation of cytokinins. Although plant regeneration through somatic embryogenesis is possible, the protocol is yet to be perfected. In this study, nodular callus was obtained from 7-9 months old immature zygotic embryos of coconut on a medium containing 24 uM 2,4-dichlorophenoxy acetic acid (2,4-D). As a novel approach, abscisic acid (ABA) at a concentration of 2.5-7.5 uM was incorporated into the culture medium for 3-7 weeks to induce somatic embryogenesis. Alternately, callus was subcultured at 5 weekly intervals on media containing gradually reducing concentrations of 2,4-D to induce somatic embryogenesis. Incorporation of ABA enhanced the production of somatic embryos. Application of 2.5-5 uM ABA for 5 weeks was found to be effective. A large number of somatic embryos developed on media containing ABA formed normal shoots and complete plants as compared to those produced in the media with low levels of 2,4-D. PMID- 10808076 TI - Adenosine, the imperfect endogenous anti-ischemic cardio-neuroprotector. PMID- 10808077 TI - Lesion of the bed nucleus of the stria terminalis enhances learned despair. AB - The bed nucleus of the stria terminalis (BNST) was lesioned in adult male Wistar rats (n = 9) and its involvement in coping behavior during forced swim stress examined. Rats remain immobile longer in the second of two swim tests, a phenomenon known as learned despair. Results revealed that, compared to sham operated controls (n = 8), BNST-lesioned animals displayed immobility significantly earlier and for longer durations in the second swim test. Rats with BNST lesions also showed significantly reduced escape behavior in the form of fewer numbers of jumps and dives compared to controls. Mobility deficits were not due to general motor impairment as revealed by an open field test. Results suggest that the BNST may modulate coping behavior especially during uncontrollable stress. PMID- 10808078 TI - Kainic acid induces distinct types of epileptiform discharge with differential involvement of hippocampus and neocortex. AB - Systemic administration of kainic acid (KA), an excitatory amino acid agonist, provides a model of epilepsy due to increased neural excitation. We examined discharges using multi-channel EEG recording and spectral analysis in rats implanted with neocortical and hippocampal electrodes after intravenous infusion of KA (10 mg/kg), until and including the first convulsive seizure. Gamma activity (30-80 Hz) increased in hippocampus from 3-9 min after KA administration. Two types of preconvulsive bilateral rhythmic discharges were observed, both consisting of generalised high voltage sharp waves at low frequencies (<10 Hz) mixed with fast oscillations (<20 Hz): (1) generalised non convulsive discharges (GNCD) occurred in all animals and (2) spike-wave discharges (SW), predominantly localised in neocortex, occurred in 45% of animals. Convulsive seizure evolved out of a GNCD. Spectral profiles of epileptiform discharges were characterised by an increase in power of low (<10 Hz) and high (beta and gamma range, 20-80 Hz) frequencies which were differently expressed in neocortex and hippocampus. Thus, in this model of convulsive epilepsy caused by increased excitation, there is an early increase in gamma activity, a process that might contribute to synchronisation, and two distinct types of bilateral discharges, hippocampal-neocortical (GNCD) and preferentially neocortical (SW). Neocortical, not hippocampal, changes in EEG power correlated with development of convulsive behaviours. PMID- 10808079 TI - Blocking the trigeminal EPSP in rat abducens motoneurons in vivo with the AMPA antagonists NBQX and GYKI 53655. AB - In pentobarbitone-anaesthetized rats, the effects of two AMPA receptor antagonists, the competitive antagonist 2, 3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F)-quinoxaline (NBQX) and the non-competitive 2,3-benzodiazepine GYKI 53655, were compared on excitatory synaptic transmission of trigeminal origin in intracellularly-recorded abducens motoneurons. The effects of both antagonists were also investigated on the alpha-amino-3-hydroxy-5-methyl isoxazole-4 propionic acid (AMPA)-, kainate-, and N-methyl-D-aspartate (NMDA)-induced depression of extracellular antidromic field potentials in the abducens motor nucleus. Microiontophoretic application (< or =100 nA) or intravenous injection of NBQX (< or =5 mg/kg) affected both AMPA- and kainate-induced depressions whereas GYKI 53655 (< or =100 nA; < or =4 mg/kg) blocked only the AMPA-induced depression. Neither NBQX or GYKI 53655 affected NMDA-induced depressions of antidromic field potentials. Using low intravenous (i.v.) doses of the antagonists NBQX or GYKI 53655 (2-2.5 mg/kg), a complete blockade of the composite disynaptic trigeminal excitatory post-synaptic potential (EPSP) was obtained without any changes in membrane potential, input resistance and antidromic action potentials in abducens motoneurons. GYKI 53655 was more potent at low i.v. doses (0.5-1.8 mg/kg) but NBQX had longer-lasting effects. The results show the existence of differences between the blocking action of NBQX and GYKI 53655 on AMPA-mediated receptor EPSP in abducens motoneurons. PMID- 10808080 TI - Glial cell line-derived neurotrophic factor in the rat pituitary gland. AB - The presence of glial cell line-derived neurotrophic factor (GDNF) is described within specific regions of the adult rat pituitary gland. Immune staining methods revealed a small number of GDNF-immunopositive cells in the anterior lobe, and in areas of the neural lobe, while no immunoreactive endocrine cells were observed in the intermediate lobe. In the neural lobe, immunofluorescence methods were also used to demonstrate that GDNF and glial fibrillary acidic protein (GFAP) are co-localized in the glial cells (pituicytes) of the neural lobe. GDNF was not co localized with neurofilament (NF) in nerve fibers of the neural lobe, suggesting that it is not present in axonal fibers. Measurements of GDNF content in separated anterior and neurointermediate lobes were also performed, using an enzyme-linked immunoassay (ELISA). Values for GDNF were slightly higher in the neurointermediate lobe than those obtained for the anterior lobe. The presence of GDNF in areas of the pituitary is discussed in the context of its possible function to support and maintain hypothalamic innervation, as well as a potential autocrine factor within endocrine cells. PMID- 10808081 TI - Local injection of pertussis toxin attenuates morphine withdrawal excitation of rat supraoptic nucleus neurones. AB - Morphine inhibits oxytocin neurones via G(i/o)-protein-linked mu-opioid receptors. Following chronic morphine administration oxytocin cells develop dependence, shown by withdrawal excitation after administration of the opioid antagonist, naloxone. Here, inactivation of G(i/o)-proteins by pre-treatment of morphine-dependent rats with pertussis toxin injected into the left supraoptic nucleus reduced withdrawal-induced Fos protein expression within the injected nucleus by 41+/-10% compared to the contralateral nucleus, indicating that functional G(i/o)-proteins are essential for the development and/or expression of morphine dependence by oxytocin cells in the supraoptic nucleus. In another group of rats, pertussis toxin did not alter the responses to either systemic cholecystokinin administration or systemic hypertonic saline administration, indicating that pertussis toxin does not prevent oxytocin cells from responding to stimuli that are not mediated by G(i/o)-proteins. Finally, pertussis toxin reduced acute morphine inhibition of systemic hypertonic saline-induced Fos protein expression in the supraoptic nucleus, confirming that pertussis toxin effectively inactivates G(i/o)-proteins in the supraoptic nucleus. Thus, the expression of morphine withdrawal excitation by supraoptic nucleus oxytocin cells requires the functional integrity of G(i/o)-proteins within the nucleus. PMID- 10808082 TI - Chronic intracerebroventricular insulin attenuates the leptin-mediated but not alpha melanocyte stimulating hormone increase in sympathetic and cardiovascular responses. AB - The co-existence of hyperinsulinemia and hyperleptinemia of obesity is well established. Additionally, both insulin resistance and leptin resistance are also characteristic of these states. Possible central nervous system (CNS) mechanisms could mediate these responses in that leptin receptors are located on hypothalamic neurons that coexpress neuropeptide-Y (NPY) or proopiomelanocortin (POMC) and both peptides that have been implicated as mediators of the CNS action of leptin. Leptin has been demonstrated to decrease or down regulate NPY expression and increase POMC expression. Insulin also has been demonstrated to decrease NPY and insulin insufficiency is associated with an increased POMC. Since both leptin and insulin share and modulate the same effector systems, we investigated the effect of CNS-induced hyperinsulinemia on the subsequent cardiovascular and sympathetic nervous response to leptin. Normal rats were implanted with intracerebroventricular (i.c.v.) cannula and allowed to recover. They were treated with insulin via i.c.v. cannula for 3 days. Following treatment, they were instrumented for the recording of cardiovascular and sympathetic nervous responses. Intracerebroventricular leptin administration in normal animals result in a progressive increase in both lumbar sympathetic nerve activity and mean arterial pressure. However, in animals pretreated with insulin for 3 days the leptin-induced response was completely attenuated. However, insulin treatment did not affect the POMC peptide product, alpha-melanocyte stimulating hormone (alphaMSH), mediated sympathetic nervous and cardiovascular responses. From these studies we conclude that CNS hyperinsulinemia can act to attenuate the leptin-induced increases in sympathetic nervous and cardiovascular system activity. The decreased responsiveness is not due to decreased sensitivity of the melanocortin, alphaMSH, mediated pathway.We suggest that the hyperinsulinemia of obesity may play a role in the obesity-induced leptin resistance. PMID- 10808083 TI - Kainate receptor-mediated activation of the AP-1 transcription factor complex in cultured rat cerebellar granule cells. AB - The sequence-specific DNA-binding activity of the AP-1 transcription factor complex was measured in cultured rat cerebellar granule cells by electrophoretic mobility shift assay. A low concentration of kainate (KA; 10 microM), but not alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA; 10 microM), enhanced DNA-binding of the AP-1 transcription factor in cultures pretreated with Concanavalin A (Con A), to prevent KA receptor desensitization. In the presence of cyclothiazide (an inhibitor of AMPA receptor desensitization), KA (10 microM) caused only a slight increase of AP-1 DNA-binding, in contrast to the threefold enhancement produced by AMPA (10 or 30 microM) or by a higher concentration of KA (30 microM), suggesting that the effect of KA, in the presence of Con A, is mediated by activation of putative KA receptors. To confirm this, the effects of the AMPA receptor-selective, non-competitive antagonist, 1-(4-aminophenyl)-3 methylcarbamoyl-4-methyl-3,4-dihydro-7, 8-methylenedioxy-5H-2,3-benzodiazepine (GYKI 53655; 50 microM), the mixed AMPA/KA receptor competitive antagonist, 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX; 50 microM), and the AMPA and GluR5 KA receptor competitive antagonist, (-)(3S,4aR,6R, 8aR)-6-[2-(1(2)H-tetrazole-5 yl)ethyl]-1,2,3,4,4a,5,6,7,8, 8a-decahydroisoquinoline-3-carboxylic acid monohydrate (LY 326325; 100 microM), were examined on AMPA- and KA-induced AP-1 activation, respectively. Our results suggest that stimulation of native KA receptors is responsible for the observed KA-specific activation of the AP-1 transcription factor complex. PMID- 10808084 TI - Opioid regulation of gonadotropin release: role of signal transduction cascade. AB - The present investigation elucidates the opioidergic modulation of gonadotropin releasing hormone release mechanism by signal transduction cascade in discrete brain regions from estrogen-progesterone primed ovariectomized rats. The effects of mu-opioid agonist morphine and its antagonist naloxone followed by morphine were studied (in two different groups of rats) on protein kinase A, adenosine 3',5' cyclic monophosphate, protein kinase C and calcium/calmodulin protein kinase-II as well as phospholipase C, phospholipase A(2), diacylglycerol and inositol 1,4, 5-triphosphate. Significant decline in phosphoinositide metabolism was observed after morphine treatment as depicted by decrease in phospholipase C and phospholipase A2 activities as well as inositol 1,4,5-triphosphate and diacylglycerol contents from discrete brain regions. Protein kinase A activity showed translocation from membrane bound to cytosolic form along with a decrease in its activator adenosine 3',5'-cyclic monophosphate levels in morphine-treated group. Calcium/calmodulin dependent protein kinase II activity also declined, whereas, protein kinase C activity increased in the cytosolic fraction after 45 min of morphine administration. Naloxone was seen to counteract the changes induced by morphine in most of the brain regions studied. Morphine also suppressed luteinizing hormone levels, whereas, follicle stimulating hormone level did not change. The present investigation provides evidence for opioidergic mediated suppression of gonadotropin release through the downregulation of signal transduction cascade. PMID- 10808085 TI - Differential potentiative effects of glutamate receptor antagonists in the production of antinociception induced by opioids administered intrathecally in the mouse. AB - The effect of (+/-)-5-methyl-10,11-dihydro-5H-dibenzo(a,d) cyclohepten-5, 10 imine maleate (MK-801) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) injected intrathecally (i.t.) on the inhibition of the tail-flick response induced by morphine, D-Ala(2)-NmePhe(4)-Gly-ol-enkephalin (DAMGO), beta-endorphin, D Pen(2,5)-enkephalin (DPDPE), or ?(trans-3, 4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl) cyclohexyl] benzeocetamide)? (U50, 488H) administered i.t. was studied in ICR mice. The i.t. injection of MK-801 (2 microg) or CNQX (1 microg) alone did not affect the basal tail-flick response. Morphine (0.2 microg), DAMGO (0.8 ng), beta-endorphin (0.1 microg), DPDPE (0.5 microg) or U50, 488H (6 microg) caused only slight inhibition of the tail-flick response. CNQX injected i.t., but not MK-801, enhanced the inhibition of the tail-flick response induced by i.t. administered morphine, DAMGO, DPDPE or U50, 488H. However, CNQX or MK-801 injected i.t. was not effective in enhancing the inhibition of the tail-flick response induced by beta-endorphin administered i.t. The potentiating effect of CNQX on tail-flick inhibition induced by morphine, DAMGO, DPDPE or U50, 488H was blocked by naloxone (from 1 to 20 microg), yohimbine (from 1 to 20 microg) or methysergide (from 1 to 20 microg) injected i.t. in a dose-dependent manner. Our results suggest that the blockade of AMPA/kainate receptors located in the spinal cord appears to be involved in enhancing the inhibition of the tail-flick response induced by stimulation of spinal mu-, delta-, and kappa-opioid receptors. Furthermore, this potentiating action may be mediated by spinal noradrenergic and serotonergic receptors. However, N-methyl-D-aspartate receptors may not be involved in modulating the inhibition of the tail-flick response induced by various opioids administered spinally. PMID- 10808086 TI - Antioxidants prevent elevation in [Ca(2+)](i) induced by low extracellular magnesium in cultured canine cerebral vascular smooth muscle cells: possible relationship to Mg(2+) deficiency-induced vasospasm and stroke. AB - Low serum concentrations of Mg(2+) ions have been reported, recently, in patients with coronary disease, atherosclerosis and stroke as well as in patients with cerebral hemorrhage. The aim of the present study was to determine whether potent antioxidants [alpha-tocopherol and pyrrolidine dithiocarbamate (PDTC)] can prevent or ameliorate intracellular Ca(2+) ([Ca(2+)](i)) overload associated with cerebral vascular injury induced by low extracellular free Mg(2+) ([Mg(2+)](o)). Exposure of cultured canine cerebral vascular smooth muscle cells to low [Mg(2+)](o) (0.15-0.6 mM) vs. normal [Mg(2+)](o) (1.2 mM) for either 10 min or 2 h induced concentration-dependent rises in [Ca(2+)](i). Treatment of the cultured cells with either PDTC (0.1 microM) or alpha-tocopherol (15 microM) for 24 h, alone, failed to interfere with basal [Ca(2+)](i) levels. However, preincubation of the cells with either alpha-tocopherol or PDTC for 24 h completely inhibited the elevation of [Ca(2+)](i) induced by exposure to low [Mg(2+)](o), not only for 10 min, but also for 2 h. These results indicate that alpha-tocopherol and PDTC prevent rises in [Ca(2+)](i) produced by low [Mg(2+)](o), which probably result from low [Mg(2+)](o)-induced lipid peroxidation of cerebral vascular smooth muscle cell membranes. Moreover, these new results suggest that such protective effects of alpha-tocopherol and PDTC on cerebral vascular cells might be useful therapeutic tools in cerebral vascular injury associated with low [Mg(2+)](o) and accumulation of [Ca(2+)](i). PMID- 10808087 TI - Bacteriological warfare amongst cats: what have we learned about cat bite infections? AB - Cat bite infections are one of the most common infectious diseases presenting to veterinary practices and to emergency rooms at human hospitals. This review describes the disease in humans and cats, the origin of organisms involved in cat bite abscesses and the importance of selected organisms such as members of the genus Porphyromonas in the disease. It also discusses future directions, the importance of identifying significant organisms and why an understanding of antimicrobial susceptibility patterns is of consequence to the outcome of the disease in humans and cats. PMID- 10808088 TI - Proteolytic inactivation of simian-11 rotavirus: a pilot study. AB - In an effort to develop alternate disinfectants for rotaviruses, pilot studies were conducted to determine if bacterial proteases could render simian-11 (SA-11) rotavirus non-infectious. SA-11 was found to be fairly temperature resistant, retaining a low-level of infectivity following 65 degrees C treatment for 2h at pH 8.5. It also resisted pH 8.5-5 at 45 degrees C for 2h. Alcalase, an alkaline protease, was the most effective of the various proteases (alcalase, durazym, neutrase, and savinase) tested. To analyze specific parameters for alcalase, SA 11 virus (10(5.5) median tissue culture infective dose/ml original titer) was treated at pH 6, 25 and 15 degrees C (simulated field conditions), with 0.1 and 1.0% alcalase. At pH 6.0 and 15 degrees C, 0.1% alcalase reduced SA-11 titer by 0. 75 log in 24h, and by 1.25 log in 120h. At 25 degrees C and pH 6.0, 0.1% alcalase reduced the titer by 2.25 log after 24h, and by 2.75 log in 120h. At pH 6.0 and 15 degrees C, 1% alcalase reduced SA-11 titer by 1.50 log in 24h and by 1.75 log in 120h. At the same enzyme concentration and pH, but at 25 degrees C the titer was reduced by 2. 75 log in the first 24h and by 3.25 log at 120h. These results show that the alkaline protease alcalase is capable of inactivating SA-11 virus to a certain degree depending on conditions. Further definition of operating parameters, demonstration of inactivation under field conditions and analysis whether the demonstrated degree of inactivation would decrease calf morbidity and mortality remain to be explored at this time. PMID- 10808090 TI - Subdivision of Salmonella enterica serovar enteritidis phage types PT14b and PT21 by plasmid profiling. AB - We have shown that plasmid profiling is a sensitive method for further identification of strains of Salmonella enterica serovar enteritidis (S. enteritidis) phage type PT21 and to a lesser extent the strains of phage type PT14b. Five and three plasmid types were identified within 33 strains of phage type PT21 and 19 strains of phage type PT14b, respectively. Plasmid types in strains of phage type PT21 showed significant correlation with geographical origin of the strain. In strains of phage type PT14b a single isolate predominated suggesting that the plasmid designated as 'C' can be directly linked with S. enteritidis PT14b strains. Application of IS200 fingerprinting did not reveal any other differences and showed just one copy of IS200 in all the 52 analysed strains. All the strains were tested for antibiotic resistance and only four strains were resistant to ampicillin, cefotaxime, cefuroxime and cotrimoxazole. This indicates that low molecular weight plasmids in Salmonella enteritidis are not responsible for the spread of antibiotic resistance. PMID- 10808089 TI - Molecular epidemiological studies on foot-and-mouth disease type O Taiwan viruses from the 1997 epidemic. AB - Sequence diversity was assessed of the complete VP1 gene directly amplified from 49 clinical specimens during an explosive foot-and-mouth disease (FMD) outbreak in Taiwan. Type O Taiwan FMD viruses are genetically highly homogenous, as seen by the minute divergence of 0.2-0.9% revealed in 20 variants. The O/HCP 0314/TW/97 and O/TCP-022/TW/97 viral variants dominated FMD outbreaks and were prevalent in most affected pig-raising areas. Comparison of deduced amino acid sequences around the main neutralizable antigenic sites on the VP1 polypeptide showed no significant antigenic variation. However, the O/CHP-158/TW/97 variant had an alternative critical residue at position 43 in antigenic site 3, which may be due to selective pressure in the field. Two vaccine production strains (O1/Manisa/Turkey/69 and O1/Campos/Brazil/71) probably provide partial heterologous protection of swine against O Taiwan viruses. The type O Taiwan variants clustered in sublineage A1 of four main lineages in the phylogenetic tree. The O/Hong Kong/9/94 and O/1685/Moscow/Russia/95 viruses in sublineage A2 are closely related to the O Taiwan variants. The causative agent for the 1997 epidemic presumably originated from a single common source of type O FMD viruses prevalent in neighboring areas. PMID- 10808091 TI - Simultaneous detection and strain differentiation of Mycobacterium bovis directly from bovine tissue specimens by spoligotyping. AB - Culture of Mycobacterium bovis is used routinely to support field diagnosis of bovine tuberculosis; however, this method is slow. Rapid detection and strain typing of M. bovis directly from 37 lesioned bovine lymph node specimens was performed by the polymerase chain reaction (PCR) based method, spoligotyping. Mycobacterial DNA was extracted from the specimens using a nucleic acid sequence capture technique. Two sets of specimens were tested, the first set comprising 16 decontaminated tissue homogenates from lesioned lymph node specimens which had been processed for BACTEC culture and a second set of 21 non-decontaminated lesioned lymph node specimens. Both sets of specimens had been frozen before analysis. Sequence capture PCR enabled detection and strain-typing of M. bovis directly from 15 of the 16 decontaminated homogenates and all 21 of the non decontaminated tissues. Four spoligotype (ST) patterns were obtained from each set; ST1, ST2, ST3 and ST16 were detected in the decontaminated specimens and ST1, ST2, ST11 and ST14 in the non-decontaminated specimens. For both sets of specimens, ST1 was the predominant strain type detected. ST patterns obtained from the BACTEC cultures of the decontaminated specimens were in agreement with those obtained directly from the tissue. The sensitivity of detection by sequence capture-PCR compared very favourably with that of BACTEC culture. ST patterns were obtained directly from tissues of 34 of the 35 culture positive specimens and the two culture negative specimens. DNA extraction from the 21 non decontaminated specimens involved an initial stomaching treatment. An assessment of sequence capture on both liquid alone and liquid and tissue homogenate combined, following stomaching, indicated that PCR was less successful on the liquid component alone. PMID- 10808092 TI - Distribution of capsular types and production of muramidase-released protein (MRP) and extracellular factor (EF) of Streptococcus suis strains isolated from diseased pigs in seven European countries. AB - Streptococcus suis strains (n=411), isolated from diseased pigs in seven European countries were serotyped using specific antisera against serotype 1 to 28, and were phenotyped on the basis of their muramidase-released-protein (MRP) and extracellular-factor protein (EF) production. Overall, S. suis serotype 2 appeared to be most prevalent (32%), followed by serotype 9 (20%) and serotype 1 (12%). Serotype 2 was most frequently isolated in France, Italy and Spain, whereas serotype 9 was most frequently isolated in Belgium, The Netherlands and Germany. In the United Kingdom serotypes 1 and 14 were most frequently isolated. High percentages of S. suis serotype 1, 2, 1/2 and 14 strains, isolated from tissues associated with S. suis infections such as brain, serosa, joint, heart and organs expressed the EF-protein, indicating that in these serotypes expression of EF is likely to be associated with virulence. In contrast, strains belonging to serotype 7 and 9, isolated from tissues associated with S. suis infections did not produce EF. These results strongly suggest that in the serotypes 7 and 9 EF expression is not related to virulence. More than 80% of the S. suis serotype 9 strains produced an MRP* protein, a high molecular variant of the 136kDa MRP. Expression of MRP* in serotype 9 strains is possibly associated with virulence. PMID- 10808093 TI - A spirochete isolated from a case of severe virulent ovine foot disease is closely related to a Treponeme isolated from human periodontitis and bovine digital dermatitis. AB - The isolation of spirochetes from severe ovine foot disease has been reported recently by our research group. In this study we describe the preliminary classification of this spirochete based on nucleotide sequence analysis of the PCR-amplified 16S rRNA gene. Phylogenetic analysis of this sequence in comparison with other previously reported 16S rRNA gene sequences showed that the spirochete belonged to the treponemal phylotype Treponema vincentii which has been associated with bovine digital dermatitis and human periodontal disease. Further work is required to define the common virulence determinants of these closely related treponemes in the aetiology of these tissue destructive diseases. PMID- 10808094 TI - Adsorption of Salmonella enteritidis by cetylpyridinium-exchanged montmorillonite clays. AB - Recent experiments in our laboratory have suggested that certain montmorillonite clays, when exchanged with the cationic surfactant cetylpyridinium (CP), may be useful in removing bacteria from aqueous solution. During an initial study, screening various CP-exchanged products for potential antibacterial activity, three CP-exchanged clays - CP*AAM (acid-activated montmorillonite), CP*STx-1 (Ca(++)-montmorillonite), and CP*SWy-2 (Na(+)-montmorillonite), proved to be the most effective. Binding studies were performed using 1mg each of CP-exchanged AAM, STx-1, and SWy-2 with a standardized Salmonella enteritidis solution containing approximately 40,000 colony forming units (CFU)/ml. The modified clays reduced bacterial numbers 98.1, 97.6, and 95.2%, respectively. In contrast, the parent clays only produced reductions of 39.8, 16.9, and 16.6%, respectively. Attempts were made to desorb CP from the modified clays by washing in sterile physiological saline for 24h. The resulting wash solutions failed to produce any significant reduction in bacterial colony counts; while, the washed clays retained their full antimicrobial activity. These findings suggested that the antibacterial effect of the clays is localized on the clay surface and is not due to CP dissociating from the clay. Electron microscopy revealed that the bacteria adhered to the surface of the CP-exchanged clays, but not the parent clays. Results from timed binding studies showed that the antibacterial effect was stable over the period observed. Rates of binding were positively influenced by increasing temperature, not affected by changes in pH, and negatively influenced by the presence of organic contaminants. The mechanism by which bacterial counts are reduced may involve the enhanced hydrophobicity and affinity of the CP exchanged clay for Salmonella and the antibacterial activity of CP. PMID- 10808095 TI - Adhesion of Streptococcus gallolyticus strains to extracellular matrix proteins. AB - Fourteen pigeon Streptococcus gallolyticus strains of differing virulence, were tested for their ability to adhere to immobilised fibronectin, collagen types I, III and IV. Eight, 2 and 13 strains were able to bind fibronectin, collagen types III and IV, respectively. None of the strains adhered to collagen type I. Heat treatment, proteolytic digestion or periodate treatment reduced the binding of S. gallolyticus to fibronectin and collagen type IV, suggesting that surface receptors contain proteins and carbohydrates. Although binding to these extracellular matrix proteins can play a role in the pathogenesis of streptococcosis in pigeons, binding properties could not be related to virulence, indicating that other factors determine differences in virulence among pigeon S. gallolyticus strains. Adhesion to collagen type IV may account in part for the distribution pattern of the lesions observed in naturally and experimentally infected pigeons. PMID- 10808096 TI - DNA commission of the international society for forensic genetics: guidelines for mitochondrial DNA typing. PMID- 10808097 TI - Pulmonary edema in fatal heroin overdose: immunohistological investigations with IgE, collagen IV and laminin - no increase of defects of alveolar-capillary membranes. AB - Pulmonary edema complicating heroin overdosage is a well recognized entity and regarded as the major mechanism contributing to death in heroin addicts. It's pathogenesis is unknown, several mechanisms are discussed: hypoxia-induced increase of pulmonary capillary permeability, depressed myocardial contractility, centrally induced respiratory depression, primary toxic effects on the alveolar capillaries and acute anaphylactic shock. The present study included opiate related deaths (n=23) and a control group of sudden cardiovascular deaths (n=12) to verify the hypothesis, that defects of the alveolar capillary membranes and/or an acute anaphylactic reaction leads to pulmonary congestion, edema and hemorrhages. Lung specimens were obtained from these 35 autopsies of persons autopsied in the Institute of Forensic Medicine, University of Bonn, in 1997 and 1998. All specimens were examined with hematoxylin-eosin, prussian blue and investigated with immunohistological methods using primary antibodies against collagen IV, laminin and IgE. Defects of the basal laminae of the alveoli were found, demonstrated by laminin and collagen IV, and the number of IgE-positive cells was counted in both groups. There was an increased but not significant number of IgE-positive cells in the heroin-group and defects of the epithelial and endothelial basal laminae were found in both groups without significant differences. PMID- 10808098 TI - Approaches to chronological age assessment based on dental calcification. AB - The aim of the present study was to find an accurate estimation of chronological age using a small number of selected teeth. For this purpose, the method devised by Nolla [C. Nolla, The development of the permanent teeth, J. Dent. Child. 27 (1960) 254-266.] was used: the development of each of the teeth was determined according to this method on 374 radiographs, 195 of boys (mean age 8.59) and 179 of girls (mean age 8.75). The 28 variables representing the calcification stages were analyzed using cluster analysis followed by multivariate analysis (multiple linear regression model). Patient age was considered to be a dependent variable. Our study showed that antimere teeth are the most homogeneous as regards stages of development. The prediction was more accurate for boys and girls below 10 years of age, using teeth 21, 43 and 46 from boys and teeth 21, 46 and 47 from girls. These teeth accounted for 80% total variance of chronological age for dental calcification. Standard error was +/-1.4 years for boys and +/-1.2 years for girls. When the age of the children remained completely unknown, the best estimates were provided by teeth 43, 47, 46 and 44 from boys and teeth 44, 47 and 43 from girls. PMID- 10808099 TI - A simple and cost-effective method for preparing DNA from the hard tooth tissue, and its use in polymerase chain reaction amplification of amelogenin gene segment for sex determination in an Indian population. AB - The use of teeth as an important resource in the analysis of forensic case history by polymerase chain reaction (PCR) or other related methods has been reported. However, a major drawback in using teeth has been that the DNA is present only in trace amounts, and the methods to recover DNA from the flinty material have not been efficient or cost effective. In this report, we describe a method to prepare DNA from the hard tooth tissues. Our studies show that ultrasonication of teeth samples yields sufficient amounts of good quality DNA useful for PCR-based diagnostic methods. The teeth could serve as a reliable source of DNA for amplification-based forensic methods in sex determination. DNA could be obtained from any tooth, regardless of the age of subject. Furthermore, by using the AMEL gene-based primers in PCR, we have shown that the AMEL gene serves as a good marker for sex determination in the Indian population. In our study, the PCR-based method was sensitive and proved to be successful for sex determination with a complete specificity. PMID- 10808100 TI - Distribution of Y-chromosome STR defined haplotypes in Iberia. AB - Seven Y-specific STR loci (DYS19, DYS389I, DY5389II, DYS390, DYS391, DYS392 and DYS393) were studied in five populations from the Iberian Peninsula: Andalusia, Valencia, Basque Country, Galicia and Northern Portugal. Haplotype and allele frequencies of these seven Y-chromosome STRs were estimated. Observed haplotype diversities are in a range between 0.96 (Basque Country) and 0.99 (Valencia and Andalusia). Significant population differentiation was registered between Basques and all the other Iberian populations and also between Valencia and Northern Portugal. PMID- 10808101 TI - Alcohol in a series of medico-legally autopsied deaths in northern Norway 1973 1992. AB - It is well established that use of alcohol increases the risk of fatal injuries. The presence of blood alcohol in autopsied deaths is regularly encountered in medico-legal practices. The aim of this study was to investigate the prevalence and concentration of alcohol in 1539 medico-legal autopsies in two counties in northern Norway in the period 1973-1992, and the reporting of acute alcohol influence among these deaths to the official cause-of-death statistics. Blood alcohol concentration (BAC) >/=0.5 per thousand (50 mg/100 ml) was found in 47.6% (n=456) of violent deaths tested, and in 93% (n=426) of these the BAC was >/=1.0 per thousand. In 17.4% (n=55) of tested natural deaths the BAC was >/=0.5 per thousand. Acute alcohol-influenced violent deaths were under-reported to the cause-of-death statistics. Deaths by motor vehicle traffic accidents did not differ from other violent deaths in this respect. The under-reporting among violent deaths was 41% in cases with BAC >/=0. 5 per thousand and 37% where the BAC was >/=1.0 per thousand during the whole period. It is concluded that post mortem BAC >/=0.5 per thousand, should be regarded as a possible contributory cause in all violent deaths, and reported accordingly. PMID- 10808102 TI - Significance of various analytical methods with reference to the causes and manners of death in alcoholics. AB - It was the aim of the present investigation to apply a broad spectrum of analyses to forensic autopsies of alcoholics in order to estimate the significance of the various analytical methods with reference to the cause and manner of death. The analyses were performed on a consecutive series of 73 medico-legal autopsies in alcoholics. Both extensive histology as well as toxicology and microbiology were used. The microbiology did not contribute substantially to the determination of the cause of death, while histology was decisive in six cases. Toxicology analyses were necessary for determining the cause of death in 37 cases. The results of the investigation may help in the selection of analytical priorities. PMID- 10808103 TI - Analytical method for the determination of strychnine in tissues by gas chromatography/mass spectrometry: two case reports. AB - This paper describes an analytical method for strychnine determination in biological samples by gas chromatography/mass spectrometry and their application in the investigation of two cases involving strychnine ingestion: A fatal case and a clinical one. The strychnine is isolated from biological samples using a liquid-liquid extraction procedure. The clean-up procedure is performed using an acid solution. Papaverine is used as internal standard in the quantification of strychnine. In the analysed specimens, the limits of quantification were 0.1 microg/ml or 0.1 microg/g. The recovery rate ranged from 75.0% to 98.7% and the coefficients of variation ranged from 4.8% to 10.5%. PMID- 10808104 TI - Aging effects upon smooth pursuit induced by step-ramp stimulation. AB - OBJECTIVE: in predictable target movements, pursuit gains are extremely close to 1.0. Under these conditions, aging effects upon pursuit have not been detected. Step-ramp stimuli produced by unpredictable combinations would be favorable procedures to assess the properties of smooth pursuit between young and aged people. METHODS: the target was a 0.5 degrees red laser spot. Eye movements of 49 normal subjects (26 younger subjects less than 49 years old and 23 older subjects more than 50 years old) were recorded with infrared reflection oculography and sampled at 250 Hz. Step direction and distance (2, 4, 6, 8 degrees) followed by 10 degrees /s ramp speed movement in the same (onward stimulus) or opposite to the direction (backward stimulus) were programmed in an unpredictable way before each set of ten tests. Both onward and backward stimulation were randomly given for each subject. RESULTS: in the onward study, there was a close correlation between retinal slip velocity (RSV) and eye acceleration (EA), which was expressed as a regression curve, being steeper in the younger group. This means that the younger subjects could produce faster velocity and greater acceleration than the aged group (P<0.05). CONCLUSION: subtle reduction in visual acuity, visual fields and contrast sensitivity will be one of possible causes, and the degradation of cortical areas crucial for visual processing might be one of possible causes of slowing of pursuit EA. PMID- 10808105 TI - Translabyrinthine removal of large acoustic neuromas in young adults. AB - OBJECTIVE: the authors reviewed the clinical manifestations and the surgical outcomes in a series of young patients who underwent removal of large acoustic neuromas via the translabyrinthine approach. METHODS: 40 young adults who underwent a translabyrinthine removal of acoustic neuromas 3 cm or greater in size were analyzed. The patient's age ranged from 17 to 30 years. The mean size of tumor was 4.25 cm. RESULTS: the primary symptoms are similar to those in adult but usually less intense. The average interval time between the primary symptom and the diagnosis was 17 months. A high percentage of preoperative normal hearing (35%) and good facial function (100%) were noted. Translabyrinthine approach was used in all cases. Total removal was realised in 39 patients (97.5%). The facial nerve was anatomically preserved in 37 patients (92.5%). Twenty-six patients (65%) had a good facial function (House-Brackmann grade I or II) immediately or at 1 month after surgery, 11 patients (28%) achieved grade III or IV. Three patients underwent an immediate nerve repair after tumor removal. All of them recovered to grade III or IV 1 year after surgery. Postoperative complications were minimal. CONCLUSIONS: young adults may have a rapid growth rate but usually have minimal symptoms even with a large acoustic neuroma. The translabyrinthine approach has been used successfully in removal of large acoustic tumors of young patients, with the good result of facial nerve preservation and long-term tumor control. PMID- 10808106 TI - A comparison and conversion table of 'the House-Brackmann facial nerve grading system' and 'the Yanagihara grading system'. AB - A comparison between the House-Brackmann facial nerve grading system (the HB system) and the Yanagihara grading system (the Y-system) was studied with 199 evaluations of 62 cases of postoperative unilateral acoustic neuroma. In the beginning, an original draft of the conversion table was formulated according to the 199 evaluations, in which, 0-6, 8-14, 16-20, 22-28, 30-38, and 40 points in the Y-system were matched with grade VI, V, IV, III, II, and I in the HB-system respectively. The result of the present study for prediction of sequelae showed that it was not necessary to consider the sequelae in a conversion table. And more, the study of an inter-observer variation showed that the lower and upper limits of the scores in the Y-system may shift within about a 2-point range in the draft table. From these aspects, a newly revised conversion table was proposed as a revised conversion table, in which, 0-6, 8-14, 16-22, 24-30, 32-38, and 40 points in the Y-system were matched with grade VI, V, IV, III, II, and I in the HB-system. This revised conversion table is much easier to remember for clinical use, because the score of the lower limit of each grade is simply in multiples of 8, that is, 0, 8, 16, 24, 32 and 40. PMID- 10808107 TI - Soluble L-selectin and interleukin-8 in otitis media with effusion. AB - OBJECTIVE: adhesion of leukocytes to vascular endothelium and their invasion into local tissues are important steps in inflammation. L-selectin plays a crucial role in leukocyte rolling and adhesion on endothelial cell surface. Interleukin-8 (IL-8) is a potent chemotactic substance toward neutrophils and lymphocytes-T and mediate their migration to local inflammation. The levels of soluble L-selectin (sL-selectin) and IL-8 were measured in middle ear effusions (MEE) from children with otitis media (OM) to estimate their role in pathogenesis of inflammation in middle ear. METHODS: MEE were collected from 113 ears of 62 children during routine myringotomy. The entire specimen was diluted with phosphate-buffered saline (PBS) to 2 ml and centrifugated at 1500 rpm for 15 min to separate cellular components. Supernatants of MEE were stored frozen at -20 degrees C for sL-selectin and at -80 degrees C for IL-8 assessment. The concentration of sL selectin and IL-8 were estimated with ELISA and compared with total protein concentration measured by Bradford assay. RESULTS: MEEs from children with chronic mucous otitis media contained significantly higher mean concentrations of IL-8 559.4 pg/mg total protein (TP) (+/-535.6) in comparison with normal serum 17.79 pg/mg TP (+/-10.9), serous OM 40.3 pg/mg TP (+/-28.1) and purulent OM 104.4 pg/mg TP (+/-128.6). High concentration of IL-8 was found in MEE from the children with early recurrence of OMS. The levels of sL-selectin were higher in purulent effusions 77.2 ng/mg TP (+/-67.4) than those of chronic mucoid 27.6 ng/mg TP (+/-36.7) and serous OM 26.3 ng/mg TP (+/-27.1). CONCLUSION: the results support a hypothesis that IL-8 can be responsible for prolongation of inflammatory process in middle ear, therefore long-term treatment and observation of children with the high levels of IL-8 in MEE may be necessary. Elevated level of sL-selectin in acute OM suggests the involvement of L-selectin in the acute phase of OM. PMID- 10808108 TI - The predictive value of auditory brainstem responses for diabetic retinopathy. AB - OBJECTIVE: The purpose was to find out whether there is a relationship between the central nervous system involvement and retinopathy in diabetes mellitus. STUDY DESIGN: In a multidisciplinary clinical study, diabetic patients with and without retinopathy were examined utilising auditory brainstem response (ABR) testing, and the results were interpreted from clinical and pathophysiological point of view. PATIENTS AND METHODS: Thirty-three diabetic patients with retinopathy (study group) and 20 diabetic patients without retinopathy (control group) were included. Audiometry and ABR testing were made. The results of both groups were compared for ABR parameters. RESULTS: Audimetric results of the study and control groups were similar. In the study group, ABR testing revealed prolonged absolute and interpeak latency of the waves. The difference was significant for I-III interval (P<0.01). The amplitudes of the waves were similar in both groups and wave V amplitude was significantly diminished in the study group. Abnormal waveforms were recorded by 40 and 12.5% in the study and control groups, respectively. CONCLUSION: Retinopathy is accompanied with lower brainstem disintegration. Microangiopathy is the possible mechanism that is involved in the occurrence of retinopathy and brainstem neuropathy. ABR can help diagnose retinopathy. Better understanding the role of ABR in diagnosis and early detection of retinopathy in diabetic patients needs further study. PMID- 10808109 TI - Postoperative results for cholesteatoma in children. AB - OBJECTIVES: to review the postoperative results for cholesteatoma in children. MATERIALS: 32 ears with acquired cholesteatoma in children operated on by a single surgeon between 1987 and 1995 and followed up more than 2 years. The mean follow-up period was 5.4 years. RESULTS: a one-stage operation was performed in seven ears (21.9%) and preplanned stage operation in 25 ears (78.1%). In the first operation, closed tympanoplasty was performed in 31 ears (96.9%) and open tymapanoplasty in one ear (3.1%). During the second stage operation, residual cholesteatoma was found in 16 ears (64.0%). Recurrent cholesteatoma was detected in 19.4% of ears treated with closed tympanoplasty. The mean postoperative air conduction hearing level was within 20 dB in 12.5%, 30 dB in 40.6% and 40 dB in 78.1%. The hearing results of type III tympanoplasty was better than those of type IV tympanoplasty. CONCLUSIONS: preplanned stage tympanoplasty is safer because of the high risk of recurrent and residual cholesteatoma. Surgical methods should be selected flexibly in individual cases depending upon cavity size, eustachian tube function and hearing level. Cholesteatoma in children should be operated on while stapes is present. PMID- 10808110 TI - Ten-year results of canal wall down mastoidectomy for acquired cholesteatoma. AB - OBJECTIVE: To examine 10-year results of canal wall down mastoidectomy (CWDM) for acquired cholesteatoma. METHODS: Medical records of 136 patients with cholesteatoma who has undergone CWDM in a university hospital and who had a follow-up of at least 10 years were checked. RESULTS: During follow-up, 21% of patients had undergone one revision operation and 3% two revisions. The recurrence rate of cholesteatoma was 17% and in three patients the cholesteatoma recurred twice. Ten years after CWDM, 98% of the operated ears were dry, 1% moist, and one ear (0.7%) was discharging. The tympanic membrane was intact in 92% and perforated in 8%. Only 14% of patients had hearing levels of 20 dB or better and 46% had 40 dB or better. CONCLUSION: It is concluded that the surgical technique of CWDM should be improved in order to lower the recurrence rate and to improve hearing results. PMID- 10808111 TI - Middle ear volume and pressure effects on tympanometric middle ear pressure determination: model experiments with special reference to secretory otitis media. AB - OBJECTIVE: Middle ear pressure (P(m)) measured by tympanometry has revealed high negative values in patients with secretory otitis media (SOM) in contrast to direct measurement. This may be explained by errors in tympanometry caused by volume displacement of the tympanic membrane (TM) affecting the volume of the middle ear (V(m)) and the P(m) according to Boyle's Law. Such errors are susceptible to the size of V(m). METHODS: A realistic middle ear model based on previous clinical studies of normal pressure-volume relations of the middle ear system (MES) was constructed. In this model non-linear behaviour and hysteresis of the MES was imitated and P(m) as well as V(m) could be controlled. RESULTS: Tympanometrically estimated P(m) decreased on average 38 daPa, when V(m) was changed from 21 to 1 cm(3). The decrease was most pronounced, when V(m) became smaller than 5 cm(3). Moreover, tympanometry showed a linear numerical overestimation of P(m) by a factor 2.31 compared with model P(m). CONCLUSION: A curve fit was derived describing the tympanometric P(m) as a function of V(m). This demonstrated that tympanometric P(m) approached -infinity daPa, when middle ear volume approached 0 cm(3), which indicates that negative tympanometric recordings and B curves can be found in ears with normal P(m) entirely due to very small V(m)'s. This explains the discrepancy between direct and tympanometric measurements of P(m) in SOM, since the effusion replaces the air filled expandable volume resulting in a very small 'functional' V(m). Numerical overestimation of P(m) by tympanometry was explained by hysteresis, which reflected the viscoelastic properties of the MES. These results question the significance of negative P(m)'s as a pathogenetic factor in SOM. PMID- 10808112 TI - Effect of photodynamic therapy on a heterotransplanted human parotid tumor. AB - To evaluate the effect of photodynamic therapy on human parotid tumors we used tumor specimens obtained from parotid surgery on a consecutive group of patients. The tumors were transplanted into a subcutaneous pocket of nude mice. The original human tumors were pleomorphic adenoma (four), adenolymphoma (one), acinic cell carcinoma (one), sarcoma (one) and low-grade adenocarcinoma (one). The most aggressive growth was seen in the low-grade adenocarcinoma. We re implanted this tumor on ten mice bilaterally, and treated the tumors with photodynamic therapy (PDT), resulting in a mean depth of tumor necrosis of 5.4 mm (1-10 mm). In three cases we found vital tumor cells in the periphery of the tumor after treatment, with several new blood vessels in the surrounding tissue, indicating a great potential for neo-angiogenesis in this tumor. In order to evaluate the possible nerve damage subsequent to the photodynamic therapy, the ischiadic nerve in 24 lower limbs of nude mice were investigated. In one case only the macroscopical and histological investigation revealed signs of nerve damage. The current study demonstrates that the nude mice implantation model is excellent to investigate growth in both malignant and benign parotid tumors, and to test new therapeutic modalities. Photodynamic therapy seems to have a possible role in the future management of the malignant lesions of the parotid gland, in cases where radical surgery for some reason is not achievable. PMID- 10808113 TI - Expression of RANTES by IL-1 beta and TNF-alpha stimulated nasal polyp fibroblasts. AB - OBJECTIVE: Accumulation of eosinophils (Eo) is one of the most characteristic feature of nasal polyps. However, the question remains why eosinophils accumulate into the nasal polyp tissue. RANTES (regulated upon activation, normal T cell expressed and presumably secreted) is a recently described chemokine that is said to play a role in the recruitment of eosinophils into inflammatory tissue sites. Fibroblasts are a rich source of cytokines and inflammatory mediators. The objective of this study was to demonstrated the expression of the chemokine RANTES in nasal polyp fibroblasts after stimulation with proinflammatory cytokines like TNF-alpha and IL-1 beta. METHODS: Fibroblast lines were established from human nasal polyp biopsy tissues taken from patients with chronic sinusitis who had no other associated diseases. Cultured nasal polyp fibroblasts were stimulated with TNF-alpha or IL-1 beta at various doses (0.1, 1.0, 1 ng/ml) or for various times (l, 6, 12, 24, 48, 72 h). To detect the RANTES gene expression, RT-PCR was performed. The resulting supernatants were assayed with ELISA for the level of RANTES. RESULTS: We demonstrated that TNF-alpha and IL-1 beta induced the gene expression and protein production of RANTES in nasal polyp fibroblasts. This responsiveness to TNF-alpha and IL-1 beta was time and dose-dependent. CONCLUSION: These findings suggest that nasal polypfibroblasts may also play an important role in the recruitment of Eo through the production of RANTES. PMID- 10808114 TI - Maxillary sinus development and sinusitis in patients with cleft lip and palate. AB - OBJECTIVE: Sinusitis is frequently associated with cleft lip and palate. The present study was performed in an attempt to investigate maxillary sinus development in relation to the occurrence of sinusitis in cleft patients. METHODS: Forty-seven consecutive patients with cleft lip and alveolus with or without cleft palate underwent computed tomographic scans of the maxilla. The cross-sectional area of the maxillary sinus at the level of the zygomatic arch was measured, and the soft tissue density shadow of the sinus was scored on a 4 point scale. RESULTS: The cross-sectional area of the sinus was significantly smaller in the child group (10 years or less) than in the adolescent group (11-20 years, P<0.001) and the adult group (over 20 years, P<0.02). The cross-sectional area significantly increased with age in the child group (r=0.552, P<0.001). The growth rate became slower in the adolescent group. The cross-sectional area then gradually decreased with age in the adult group (r=-0.64, P<0.05). Sinusitis was observed in 15 patients (32%) and in 20 sinuses (21%). Severe sinusitis was more common in the child group than in the patients over 10 years of age (P<0.05). CONCLUSION: These results indicate that maxillary sinusitis associated with cleft lip and palate occurs preferably in the developing sinuses of children. PMID- 10808115 TI - Zinc modulates the electro-olfactogram of the frog. AB - OBJECTIVE: Zinc is considered an important element in olfaction, however, its exact role is still unclear. A large amount of zinc is contained in the olfactory mucosa, so Zn(2+) may behave as a neuromodulator, as in the hippocampus. To reveal the acute effects of Zn(2+) on olfactory mucosa, electro-olfactograms (EOG) were recorded with or without Zn(2+). METHODS: The isolated olfactory mucosa of a bullfrog (Rana catesbiana) was set in an EOG recording chamber and bathed in Ringer's solution. Effects of ZnSO(4) (25 microM) on EOGs were examined. Four types of stimulants, n-amyl acetate (200 microM), menthone (10 microM), forskolin (2 microM), and 3-isobutil-methylxisanthine (IBMX, 100 microM) were tested. RESULTS: Zn(2+) (25 microM) charged with the ciliated surface of the olfactory mucosa attenuates EOGs which were produced not only by odorants but also by forskolin and IBMX. The attenuating ratios of EOGs for stimulants were as follows; n-amyl acetate (0.51+/-0.18, n=4), menthone (0.48+/-0.28, n=4), forskolin (0.61+/-0.16, n=4), and IBMX (0.68+/-0.19, n=4); (mean+/-S.D.). These attenuations were reversible. The results indicate that Zn(2+) may block the c AMP dependent transduction channels of the olfactory cells. CONCLUSION: Micro molar Zn(2+) attenuates EOG of the frog. Zn(2+) may be thought to behave as one of the modulators in olfactory reception. PMID- 10808116 TI - Conditions of the uvula: a 14 years experience. AB - OBJECTIVE: A retrospective study to find out the aetiology, symptomatology and management of elongated and enlarged uvulae. MATERIALS AND METHODS: Examination of files of 375 patients seen in the period 1983-1997. RESULTS: Trauma constituted a big proportion of patients seen. However, papillomas, solitary infections, allergy, simple symptomatic elongation, squamous cell carcinoma as well as asympomatic elongation were seen. The symptomatology varied between simple irritation in the throat to vomiting, pain and dysphagia. CONCLUSION: Enlarged or elongated uvulae are rare but solitary involvement of uvula because of variety of causes occur. PMID- 10808117 TI - Infiltrating angiolipoma of the M. temporalis. AB - Angiolipomas are rare benign mesenchymal tumours that are distinguished from common lipomas by a marked degree of vascularisation. They are differentiated into non-infiltrating and the even less frequent infiltrating angiolipomas. The present case is the 9th report of an infiltrating angiolipoma of the head and neck. The patient was a 63-year-old man with an infiltrating angiolipoma of the left M. temporalis. Microscopic examination showed univacuolated adipose cells mixed with capillaries invading skeletal muscle. The patient has been free of recurrence since excision of the tumor. PMID- 10808118 TI - A case of Meniere's disease with vertical nystagmus after administration of glycerol. AB - A 62-year-old woman who complained of repetitive vertigo with a left fluctuating hearing loss was admitted to our hospital three times. A glycerol test was done on the third admission. Two hours after the administration of glycerol, the patient complained of rotatory vertigo with a downbeat vertical nystagmus. This nystagmus then changed its direction upward. Her left hearing loss was improved during the glycerol test. On the day after the glycerol test, a caloric test was done. The caloric response of the right ear was remarkably improved. Although she had not felt a hearing loss in the right ear, an overwritten audiogram showed a fluctuation of hearing in the right ear. The bilateral caloric responses fluctuated. From these findings, it appears that the function of the inner ear on both sides fluctuating. We speculate that the administration of glycerol elicited an asymmetry in the function of the inner ear on both sides and ocuured vertigo with vertical nystagmus because of the irritation of both ears. PMID- 10808119 TI - Stapes surgery and psychiatric complications. AB - Changes in auditory perception can cause disturbances in development and personality. This phenomenon has been studied in particular in children hearing loss or in progressive or sudden hearing loss in the adult. We present the case of a patient with psychobehavioural alterations after restoration of hearing following a small fenestra stapedectomy for bilateral otosclerosis with moderate severe hearing impairment. The diagnosis, physiopathology and medicolegal implications are discussed. PMID- 10808120 TI - Squamous cell carcinoma presenting as a cutaneous horn. AB - A cutaneous horn is a variably sized, cohesive, keratotic, usually conical lesion associated with a broad spectrum of pathology at their base (benign, premalignant or malignant). Two cases of a squamous cell carcinoma in the face, with a horn formation are presented, highlighting the need for careful management of such lesions due to the high incidence of malignant or premalignant histology. The existing literature is also briefly reviewed. PMID- 10808121 TI - Creutzfelt-Jakob disease presenting hyperparathyroidism. AB - We reported a 68-year-old female with Creutzfelt-Jakob disease (CJD) presenting hyperparathyroidism. She was suspected as Creutzfelt-Jakob like syndrome at her initial visit to our hospital because of progressive dementia and high level of serum calcium. Finally she was diagnosed as having CJD by the clinical symptoms including progressive dementia and myoclonus and the characteristic patterns of brain CT and electroencephalogram (EEG). CJD presents a variety of symptoms including progressive dementia, apathy and myoclonus. Hyperparathyroidism and toxicity of lithium, delirium and bismuth have been reported to induce similar symptoms of CJD, which are called Creutzfelt-Jakob like syndrome. Therefore, the diagnosis of CJD would be difficult in cases with CJD accompanied with Creutzfelt Jakob like syndrome. It is rare for otolaryngologists to examine patients with dementia like CJD. However, we have to keep CJD in mind in the diagnosis of hyperparathyroidism. PMID- 10808122 TI - Huge osteoma of the frontoethmoidal sinus with secondary brain abscess. AB - Osteomas of paranasal sinuses are usually asymptomatic. When enlarged, they may cause serious complications. We report a case of huge frontoethmoidal osteoma associated with brain abscess. If an osteoma associated with an intracerebral pathology is detected, it should be remembered that this could have been a complication of the osteoma and a detailed radiological examination should be performed to show the extent of the osteoma and to confirm any defect at the posterior wall of the sinus which may lead intracerebral complications. Such an osteoma should be operated after the abscess was resolved and patient became stable neurologically. PMID- 10808124 TI - Beta 1-integrin-mediated cell signaling in T lymphocytes. AB - beta1-integrins play crucial roles in a variety of cell processes such as adhesion, migration, proliferation, and differentiation of lymphocytes. For understanding the molecular mechanisms of these various biological effects, it may be particularly important to analyze cell signaling through the beta1 integrins. Our previous study had shown that PLC-gamma, pp125FAK (focal adhesion kinase), pp105, paxillin, p59fyn, p56lck and ERK1/2 are phosphorylated in their tyrosine residues upon engagement of beta1-integrins. We identified pp105 as Cas (Crk-associated substrate)-related protein and successfully cloned its cDNA. pp105 is a Cas homologue predominantly expressed in the cells of lymphoid lineage, which led us to designate it as Cas-L. Like p130Cas, Cas-L contains a single SH3 domain and multiple SH2 binding sites (YXXP motif), which is suggested to bind SH2 domains of Crk, Nck, and SHPTP2. Subsequent studies revealed that pp125FAK binds Cas-L on its SH3 domain and phosphorylates its tyrosine residues upon beta1-integrin stimulation. Since Cas-L is preferentially expressed in lymphocytes, it is conceivable that Cas-L plays an important role in lymphocyte specific signals. We have shown that Cas-L is involved in the T-cell receptor (TCR)/CD3 signaling pathway as well as the beta1-integrin signaling pathway. Cas L is transiently phosphorylated following CD3 cross-linking, and tyrosine phosphorylated Cas-L binds to Crk and C3G. Furthermore, a Cas-L mutant (Cas LDeltaSH3), which lacks the binding site for FAK, is still tyrosine phosphorylated upon CD3 cross-linking, but not upon beta1-integrin cross-linking, suggesting that FAK is not involved in CD3-dependent Cas-L phosphorylation. Finally, we have identified a crucial role of Cas-L in beta1-integrin-mediated T cell co-stimulation. beta1-integrins have known to provide a co-stimulus for TCR/CD3-driven interleukin-2 production and proliferation of peripheral T-cells. We have found that this co-stimulatory pathway is impaired in the Jurkat T-cell line, and that the expression level of Cas-L is reduced in Jurkat cells compared with peripheral T-cells. The transfection of Cas-L cDNA into Jurkat cells restored the beta1-integrin-mediated co-stimulation, while the transfection of Cas-LDeltaSH3 mutant failed to do so, showing a contrast to the case with CD3 mediated signaling. These results indicate that Cas-L plays a key role through the association and phosphorylation by FAK in the beta1-integrin-mediated T-cell co-stimulation. Taken together, Cas-L might be the bi-modal docking protein that assembles the signals through beta1-integrins and TCR/CD3, and participates in a variety of T-cell functions. PMID- 10808125 TI - Lack of androgen receptor transcriptional activity in human keratinocytes. AB - Since detection of androgen receptor (AR) expression in keratinocytes by immunostaining is controversial, we investigated whether keratinocytes can act as androgen target cells using transient transfection assays. Chloramphenicol acetyltransferase (CAT) assays for the endogenous AR transcriptional activity in HaCaT keratinocytes indicated that DHT (10(-9)-10(-8) M) can induce less than 1.5 fold of mouse mammary tumor virus CAT, which is quite low, compared with 38-fold induction by 10(-7) M 1,25-dihydroxyvitamin D(3) of P450cc24-CAT. Furthermore, this low DHT-mediated induction could not be enhanced by the AR co-activators, ARA70 or ARA55. Western blotting analysis indicated that HaCaT and normal keratinocytes do not express AR protein. Transfection of exogenous AR into HaCaT keratinocytes, however, could install AR transcriptional activity, suggesting that HaCaT keratinocytes have all the necessary accessory factors for AR transcription activity. In conclusion, keratinocytes are unlikely to be target cells for androgen. PMID- 10808126 TI - Subepidermal blistering disease with autoantibodies against a novel dermal 200 kDa antigen. AB - A number of autoimmune subepidermal blistering diseases are characterized by the distinct autoantigens of the cutaneous basement membrane zone. Recently, a few cases with autoantibodies against a novel 200-kDa dermal protein have been reported. We collected nine cases of subepidermal blistering disease with IgG antibodies against this 200-kDa antigen. In this report, we describe the clinical and immunological appearances in these cases. Five cases showed bullous pemphigoid-like features, one case resembled dermatitis herpetiformis, and another case showed mixed features of bullous pemphigoid and linear IgA bullous dermatosis. It was interesting to note that psoriasis coexisted in four cases. By indirect immunofluorescence on 1 M NaCl split skin, IgG antibodies from all sera reacted with the dermal side of the split. By immunoblot analysis, IgG antibodies recognized a 200-kDa protein of dermal extract. IgG affinity-purified antibodies on the 200-kDa immunoblot membrane stained the dermal side of 1 M NaCl split skin. Various examinations suggested that the 200-kDa antigen is not identical to any chains of laminins-1, -5 or -6. This autoimmune subepidermal blistering disease against the dermal 200-kDa protein may form a new distinct entity, which often associates with psoriasis. PMID- 10808127 TI - Effects of dehydroepiandrosterone on collagen and collagenase gene expression by skin fibroblasts in culture. AB - Dehydroepiandrosterone (DHEA) and its sulfate (DHEA-S) are the most abundant steroids in humans whose low levels are related to aging, greater incidence of various cancers, immune dysfunction, atherosclerosis, and osteoporosis. It has been shown that collagen and collagenase gene expression decreases in fibroblasts taken from more aged donors. In this paper, to investigate the relationship between DHEA and skin aging, we examined the effects of DHEA on the regulation of collagen, collegians and stromelysin-1 genes in cultured human skin fibroblasts. In collagen assay, DHEA slightly increased collagen production in a dose-related fashion, its maximal effect occurred at 10(-5) M DHEA (P>0.05). In the presence of DHEA, steady-state levels of alpha1 (I) procollagen mRNA increased to 1. 6 fold of the non-treated group, while those of fibronectin were not. Interestingly, DHEA differently regulated collagenase and stromelysin-1 gene expression. The steady-state levels of collagenase mRNA decreased in response to DHEA by 40%, whereas those of stromelysin-1 mRNA increased up to 2.4-fold, compared to controls. Similar results were obtained for chloramphenicol acetyltransferase assay (CAT); maximal promoter activation of stromelysin-1 gene occurred at 10(-6) M DHEA, 4.5-fold higher than control. CAT assay revealed that treatment with 10(-5) M DHEA resulted in a strong ( approximately 70%) inhibition of the collagenase promoter activity. In our experiments, the effects of DHEA on these gene expressions were higher at pharmacologic concentration (>/=10(-5) M) than those at physiologic concentration (10(-8)-10(-6) M). This study suggests that the level of DHEA may be related to the process of skin aging through the regulation of production and degradation in extracellular matrix. PMID- 10808128 TI - Expression vector with DNA of bovine papilloma virus 1 for keratinocyte gene therapy. AB - Although there are several methods for introducing the genes to keratinocytes in vivo, expression of transgene does not last long enough for effective keratinocyte gene therapy. In this study, we added bovine papilloma virus 1 (BPV) DNA into expression vectors with the lacZ gene driven by metallothionein and keratin 10 promoters, and we transferred them into keratinocytes in vivo using the naked DNA method, and measured beta-gal activity in keratinocytes. The results showed that beta-galactosidase activity of vectors with the BPV DNA was clearly higher than that without the DNA. Moreover, time-course experiment disclosed that the activity of the BPV vector declined at a lower rate than that of the control vector, suggesting this fragment prolonged transgene expression. These results should prove useful for understanding gene regulation in keratinocytes in vivo and for developing potential expression vectors for keratinocyte gene therapy. PMID- 10808129 TI - Probable involvement of a germ-line mutation of an unknown mismatch repair gene in a Japanese Muir-Torre syndrome phenotype. AB - A combination of haplotype analysis and direct sequencing were conducted on Japanese Muir-Torre syndrome kindred. In the kindred, two females revealed a hereditary non-polyposis colon cancer (HNPCC) phenotype and one male had a sebaceous tumor in addition to a HNPCC phenotype. Haplotype analysis and direct sequencing failed to show involvement of the known mismatch repair genes, with the exception of MSH5, in this kindred. Analysis of large fragments (from 3.9 to 6. 2 kb) covering the entire 25 kb MSH5 gene in the proband revealed the absence of gross changes in the promoter region and exons. The direct sequencing of the promoter region and all 25 exons failed to demonstrate any mutations in the coding regions except for a CA repeat polymorphism in intron 3 and a C/A polymorphism in intron 15. Taken together present results indicate that a novel and yet unknown mismatch repair gene is likely involved in the HNPCC in this kindred. PMID- 10808131 TI - A new skin equivalent model: dermal substrate that combines de-epidermized dermis with fibroblast-populated collagen matrix. AB - Epidermis reconstructed on de-epidermized dermis (RE-DED) and on fibroblast populated collagen matrix (Living Skin Equivalent) showed a histologic resemblance to native epidermis. However, some abnormalities have been found including different expression pattern of differentiation markers from native epidermis. In this study, to reconstruct an epidermis model resembling native epidermis more closely than previous skin equivalents, de-epidermized dermis (DED) was raised on fibroblast-populated collagen matrix and keratinocytes were cultured on top of the DED at the air-liquid interface. The new skin equivalent like RE-DED showed a similar morphology to that of native epidermis. Immunohistochemical studies revealed that differentiation markers such as involucrin, loricrin and filaggrin but not keratin 1 expressed similar pattern characteristics to native epidermis compared with those of RE-DED. In addition, the new model showed some fibroblasts in the DED as a result of migration from the fibroblast-populated collagen matrix, mimicking a living dermis in vivo. These results indicate that the new model seems to be a better skin equivalent model than previous models. Also, they provide additional evidence that the presence of fibroblasts improves epidermal differentiation. PMID- 10808130 TI - gp100 mRNA is more sensitive than tyrosinase mRNA for RT-PCR amplification to detect circulating melanoma cells in peripheral blood of melanoma patients. AB - Two different melanocyte-specific mRNAs are studied as markers for circulating melanoma cells in vitro using the human melanoma cell line G361 and in vivo using blood samples from Japanese melanoma patients at different clinical stages. These mRNAs encode tyrosinase, the most essential enzyme for melanin synthesis, and gp100, a melanosomal matrix glycoprotein recognized by monoclonal antibody HMB 45. We used reverse-transcription polymerase chain reaction (RT-PCR) to detect tyrosinase mRNA and gp100 mRNA in peripheral blood. Since melanocytes would not normally be present in peripheral blood, the detection of those transcripts should indicate the presence of circulating melanoma cells. RT-PCR detection of these two mRNAs was highly sensitive and specific. Our in vitro study showed that as few as 10 melanoma cells in 0.125 ml normal blood could be detected. In in vivo study, 130 blood samples from 55 melanoma patients gave positive and variably sensitive results, whereas no samples from healthy controls or patients with other cancers gave positive results. Tyrosinase mRNA was not detected in any of the melanoma patients. gp100 mRNA was detected in 12 of 55 melanoma patients, in none of five stage I patients (0%), in four of 26 stage II patients (15.4%), in one of six stage III patients (16. 7%) and in seven of 18 stage IV patients (38.9%). Thus gp100 mRNA is a more sensitive marker for detecting circulating melanoma cells compared with tyrosinase mRNA. PMID- 10808132 TI - Expression of T-cell cytokines in challenged skin of murine allergic contact photosensitivity: low responsiveness is associated with induction of Th2 cytokines. AB - Murine contact photosensitivity (CPS) to 3,3',4', 5-tetrachlorosalicylanilide (TCSA) is genetically controlled mainly by the major histocompatibility complex. The H-2(b,d) haplotypes are closely associated with high responders, whereas mice with H-2(k) are low or non-responders. We found that BALB/c (H-2(d)) mice were high responders in CPS not only to TCSA but also to chlorpromazine (CPZ) and benzocaine, whereas AKR/n (H-2(k)) mice were hyporesponsive to these three photoallergic agents. To elucidate the relationship between CPS responsiveness and T helper cell induction, the expression of T-cell cytokines was examined by reverse transcriptase polymerase chain reaction in the elicited skin of CPS to the three chemicals. The expression levels of interleukin(IL)-2 and interferon gamma mRNAs were markedly higer in BALB/c mice than AKR/n mice, whereas mRNA for IL-4 was expressed strongly in AKR/n mice. These data suggest that the hyporesponsiveness of CPS to the photoallergens in AKR/n mice is closely associated with the activation/induction of Th2 cytokines at the challenged sites. PMID- 10808133 TI - Prism adaptation in normal aging: slower adaptation rate and larger aftereffect. AB - The effect of aging on prism adaptation, a motor learning paradigm, was evaluated. Different measures were obtained from a task consisting of throwing clay balls to a target in front of the subjects before, during, and after wearing prisms that deviate the visual field by several degrees. When performing the task without wearing the prisms, the aged subjects showed a larger hit variance, whereas the young subjects hit closer to the target. When donning the prisms, the aged group adapted more slowly than the controls, although after throwing all the balls both groups showed the same adaptation levels. After removing the prisms, the aged group showed a larger aftereffect. These findings suggest that the aftereffect requires the involvement of non-cognitive and cognitive processes and indicate that both adaptation and aftereffect are influenced by aging. PMID- 10808134 TI - Recognition of emotional prosody and verbal components of spoken language: an fMRI study. AB - This study examined the neural areas involved in the recognition of both emotional prosody and phonemic components of words expressed in spoken language using echo-planar, functional magnetic resonance imaging (fMRI). Ten right-handed males were asked to discriminate words based on either expressed emotional tone (angry, happy, sad, or neutral) or phonemic characteristics, specifically, initial consonant sound (bower, dower, power, or tower). Significant bilateral activity was observed in the detection of both emotional and verbal aspects of language when compared to baseline activity. We found that the detection of emotion compared with verbal detection resulted in significant activity in the right inferior frontal lobe. Conversely, the detection of verbal stimuli compared with the detection of emotion activated left inferior frontal lobe regions most significantly. Specific analysis of the anterior auditory cortex revealed increased right hemisphere activity during the detection of emotion compared to activity during verbal detection. These findings illustrate bilateral involvement in the detection of emotion in language while concomitantly showing significantly lateralized activity in both emotional and verbal detection, in both the temporal and frontal lobes. PMID- 10808135 TI - The effects of sequence structure and reward schedule on serial reaction time learning in the monkey. AB - This research tests the hypothesis that sequence learning performance in non human primates will be modulated both by the structure of the sequences to be learned and by the schedule of reward applied during learning. Sequence learning in humans has been extensively explored with serial reaction time (SRT) protocols where learning is revealed by reduced reaction times for stimuli presented in repeating sequences vs. stimuli presented in random series. The SRT protocol has been used to demonstrate that different types of sequential structure may be learned under different awareness conditions. Here, we consider surface and abstract structure of sensorimotor sequences such that sequences ABCBAC and DEFEDF (where A to F correspond to spatial locations on a touch sensitive screen) have different serial order or surface structure, but share the same abstract structure 123213, and are thus considered isomorphic. In four experiments, we manipulated the type of sequential structure to be learned, and the schedule of reward in spatial sequence learning tasks. Both of the two monkeys tested demonstrated significant SRT learning for serial order or surface structure, while they failed to learn and transfer abstract structure. Their learning performance was also modulated by the schedule of reward. These results are in support of our hypothesis and are discussed in the context of existing models of sensorimotor sequence learning. PMID- 10808136 TI - Different scalp topography of brain potentials related to expression and identity matching of faces. AB - Event-related potentials (ERPs) and behavioral data were recorded while subjects performed two tasks on the same set of faces (presented in pairs). One task was identity matching and the other expression matching. Two groups of subjects participated, one familiar and the other unfamiliar with the faces. Subjects were less accurate in matching expressions than identity. Familiarity facilitated identity but not expression matching. ERPs to mismatches in both tasks elicited a negativity around 400 ms, which was similar in latency and amplitude in the two tasks, but differed in scalp topography. Whereas the mismatch negativity had the same landscape over the left hemisphere for both tasks, the component related to expression had larger amplitudes over the right-temporal regions. Familiarity had no effect on these negativities, although it affected a late positivity (LP). These results support the idea of distinct neural systems subserving face processing, and agree with a role of the right hemisphere for the processing of emotional expressions. PMID- 10808137 TI - Internally driven vs. externally cued movement selection: a study on the timing of brain activity. AB - Brain imaging studies in man and single cell recordings in monkey have suggested that medial supplementary motor areas (SMA) and lateral pre-motor areas (PMA) are functionally dissociated concerning their involvement in internally driven and externally cued movements. This dichotomy, however, seems to be relative rather than absolute. Here, we searched for further evidence of relative differences and aimed to determine by what aspect of brain activity (duration, strength, or both) these might be accounted for. Event-related potentials (ERPs) were recorded while healthy, right-handed subjects selected one of three possible right hand digit movements based either on 'internal' choice or 'external' cues. The results obtained from ERP mapping suggest that movement selection evokes the same electrical brain activity patterns in terms of surface potential configurations in the same order and at the same strength independent of the selection mode. These identical configurations, however, differed in their duration. Combined with the results of a distributed source localization procedure, our data are suggestive of longer lasting activity in SMA during the 'internal' and longer lasting activity in PMA during the 'external' condition. Our results confirm previous findings in showing that SMA and PMA are distinctively involved in the two tasks and that this functional dichotomy is relative rather than absolute but indicate that such a dissociation can result from differences in duration rather than pure strength of activation. PMID- 10808138 TI - Neural correlates of semantic associative encoding in episodic memory. AB - Associations between individual items are the basic building blocks of learning and memory. Functional neuroimaging has now made it possible to study neural correlates of such associations. The present PET study examined three associative encoding conditions differing in the number of words (0, 1, or 2) semantically related to a third word representing the name of a semantic category. A recall task consisting in the presentation of the category names as cues for retrieving the other two members of the triads followed each encoding condition. As expected, retrieval performance increased as the number of semantic exemplars at encoding increased (10%, 43%, 70% items recalled, respectively). A first analysis (partial least squares, PLS) of the PET data identified task-related patterns of activity for associative encoding and cued-recall tasks. A second analysis identified brain regions whose activity was modulated by the number of semantic exemplars at encoding. Some of the task-related brain regions also showed modulated activity by semantic relatedness and consisted in the left inferior prefrontal cortex, right medial temporal lobe, fusiform gyrus and inferior temporal gyrus bilaterally. Some of these regions showed greater activity when words in a triad were unrelated, whereas others did so when the three words were semantically related. These regions have been consistently reported in previous functional neuroimaging studies of associative encoding and may constitute key structures in association formation. PMID- 10808139 TI - Dissociation of writing processes: functional magnetic resonance imaging during writing of Japanese ideographic characters. AB - Dissociation between copying letters and writing to dictation has been reported in the clinical neuropsychological literature. Functional magnetic resonance imaging (fMRI) was conducted in normal volunteers to detect the neurofunctional differences between 'copying Kanji', the Japanese ideographic characters, and 'writing Kanji corresponding to phonological information'. Four tasks were conducted: the copying-Kanji task, the writing-Kanji-corresponding-to-phonogram task, the Kanji-grapheme-puzzle task, and the control task. The right superior parietal lobule was extensively activated during the copying-Kanji task (a model of the copying letters process) and the Kanji-grapheme-puzzle task. These observations suggested that this area was involved in referring the visual stimuli closely related to the ongoing handwriting movements. On the other hand, Broca's area, which is crucial for language production, was extensively activated during the writing-Kanji-corresponding-to-phonogram task (a model of the writing to-dictation process). The Kanji-grapheme-puzzle task activated the bilateral border portions between the inferior parietal lobule and the occipital lobe, the left premotor area, and the bilateral supplementary motor area (SMA). Since the Kanji-grapheme-puzzle task involved manipulospatial characteristics, these results suggested cooperation between visuospatial and motor executive functions, which may be extensively utilized in demanding visual language processing. The neurofunctional difference between 'copying Kanji' and 'writing Kanji corresponding to phonogram' was efficiently demonstrated by this fMRI experiment. PMID- 10808140 TI - Neurophysiological and behavioral indices of time pressure effects on visuomotor task performance. AB - Using a video game format, this study examined the effects of time pressure (TP) on behavioral and electrocortical indices. The behavioral results were consistent with previous time pressure research in that TP reduced time to perform a task and increases behavioral errors. In addition, electroencephalogram (EEG) measures showed distinctive patterns associated with TP in the theta, mu, and gamma bands along the midline. Site specific changes in the success vs. failure trials were also seen in midline theta at Fz, gamma at Fz, and mu at Cz. Right parietal alpha also differentiated TP and success vs. failure trials. In specific TP (1) increased frontal midline theta activity and (2) increased gamma at midline (frontal, central, and partietal) and in right frontal areas. The results of these findings are discussed in terms of the formation of specific neurocognitive strategies as evidenced by the topographic distribution of task-related modulation of the EEG within certain frequency bands. It is suggested that the effect of TP on visuomotor performance is mediated by adopting either task relevant or task-irrelevant neurocognitive strategies as evidenced by successful or failed trials, respectively. Whether these strategies are formulated prior to performance or appear spontaneously during task performance remains unclear and is awaiting further experimentation. PMID- 10808141 TI - Long-range EEG synchronization during word encoding correlates with successful memory performance. AB - Distinct cortical activity during memory encoding of words, which were either recalled or not, was reported by a number of studies. This activity was mainly found at frontal and temporal/parietal brain regions. However, it was not clear if these regions interact with each other or work independently. In order to get a functional measure of the degree of neuronal large-scale cooperation, we calculated EEG coherence, which provides a statistical measure of synchronization between two EEG signals per frequency band. Therefore, coherence enables us to assess the functional interaction between cell assemblies of distant brain regions. The purpose of our study was to investigate if successfully recalled words show enhanced cortical synchronization compared with not recalled ones. Additionally, the influence of stimulus modality and the way different EEG frequencies participate in this process was examined. The EEG of 25 participants was recorded during memory encoding of concrete German nouns, either presented auditorily or visually and stimuli were separated according to the participant's memory performance. Recalled nouns exhibited overall enhanced synchronization but showed typical patterns, especially between anterior and posterior brain regions in all frequency bands except the alpha-1 band (8-10 Hz). Recalling nouns was accompanied by increased synchronization between more distant electrodes in relation to an increase of synchronization between adjacent electrodes. Moreover, the degree of intrahemispheric synchronization was higher for recalled nouns. The pattern of EEG coherence and amplitude changes during verbal memory encoding allowed us to assess the probability that nouns would be recalled or not. PMID- 10808142 TI - Increases in cortical acetylcholine release during sustained attention performance in rats. AB - Acetylcholine (ACh) efflux in the frontoparietal cortex was studied with in vivo microdialysis while rats performed in an operant task designed to assess sustained attention. Transferring animals from the baseline environment into the operant chambers elicited a robust increase in cortical ACh efflux that persisted throughout the 18-min pre-task period. Subsequent performance in the 36-min sustained attention task was associated with further significant increases in frontoparietal ACh efflux, while the termination of the task resulted in a delayed decline in ACh levels. Upon the 12-min presentation of a visual distracter (flashing houselight, 0.5 Hz) during task performance, animals initially developed a significant response bias to the left lever in the first 6 min distracter block, reflecting a reduction of attentional effort. Under continued conditions of increased attentional demand, performance recovered during the second 6-min distracter block. This return to attentional processing was accompanied by an increase in cortical ACh efflux, suggesting that the augmentation of attentional demand produced by the distracter elicited further increases in ACh release. The enhancement of cortical ACh efflux observed prior to task performance implies the presence of complex relationships between cortical ACh release and anticipatory and/or contextual factors related to operant performance and attentional processing. This finding, along with the further increases in cortical ACh efflux associated with task performance, extends hypotheses regarding the crucial role of cortical cholinergic transmission for attentional functions. Furthermore, the effects of the distracter stimulus provide evidence for a direct relationship between attentional effort and cortical ACh release. PMID- 10808143 TI - High-speed memory scanning: a behavioral argument for a serial oscillatory model. AB - In order to account for the memory span [G.A. Miller, The magical number seven, plus minus two: some limits on our capacity for processing information, Psychol. Rev. 63 (1956) 81-97.], the magical number seven, plus minus two, and high-speed scanning in human memory ?S. Sternberg, High speed scanning in human memory, Science 153 (1966) 652-654., Lisman and collaborators [O. Jensen, J.E. Lisman, An oscillatory short-term memory buffer model can account for data on the Sternberg task, J. Neurosci. 18 (1998) 10688-10699; J.E. Lisman, M.A.P. Idiart, Storage of 7+/-2 short-term memories in oscillatory subcycles, Science 267 (1995), 1512 1515.] proposed an oscillatory short-term memory buffer model. In this neurophysiological model: "a single brain network can separately maintain up to seven memories by a multiplexing mechanism that uses theta and gamma brain oscillations for clocking. A memory is represented by groups of neurons that fire in the same gamma cycle" ?O. Jensen, J.E. Lisman, An oscillatory short-term memory buffer model can account for data on the Sternberg task, J. Neurosci. 18 (1998) 10688-10699, p. 10688. To test this model, we tried to modify the memory scanning time by shifting the gamma oscillation frequency. To this aim, we replicated the visual short-term memory scanning task ?S. Sternberg, High speed scanning in human memory, Science 153 (1966) 652-654., and we simultaneously used the protocol that Treisman ?M. Treisman, A. Faulkner, P.L.N. Naish, D. Brogan, The internal clock: evidence for a temporal oscillator underlying time perception with some estimates of its characteristics frequency, Perception 19 (1990) 705 743. designed to drive, slowing down or speeding up, a temporal oscillator acting in the gamma range ?J.G.R. Jefferys, R.D. Traub, M.A. Whittington, Neuronal networks for induced "40 Hz rhythms, Trends Neurosci. 19 (1996) 202-208; W. MacKay, Synchronized neuronal oscillations and their role in motor processes, Trends Cog. Sci. 1 (1997) 176-183; M. Treisman, N. Cook, P.L.N. Naish, J.K. MacCrone, The internal clock: electroencephalographic evidence for oscillatory processes underlying time perception, Q. J. Exp. Psychol. 47A (1994) 241-289.. In this protocol, an auditory periodic stimulus (click train) was delivered at various frequencies during the task. The reaction time (RT), the slope, and the intercept of the linear function associating RT to memorized list length showed systematic modulations according to the stimulation frequency. The predicted driving effects due to the click trains were obtained, consisting of localised modulations of performance on the stimulation frequency band. We argue that memory scanning is indeed paced by a temporal oscillator, thus providing behavioral arguments for the serial oscillatory model of Lisman. PMID- 10808144 TI - Visuospatial imagery is a fruitful strategy for the digit span backward task: a study with near-infrared optical tomography. AB - Our newly developed 64-channel time-resolved optical tomographic imaging system using near-infrared light enables us to obtain a quantitative image of hemoglobin concentration changes associated with neuronal activation in the human brain ?H. Eda, I. Oda, Y. Ito, Y. Wada, Y. Oikawa, Y. Tsunazawa, M. Takada, Y. Tsuchiya, Y. Yamashita, M. Oda, A. Sassaroll, Y. Yamada, M. Tamura, Multi-channel time resolved optical tomographic imaging system, Rev. Sci. Instrum., 70 (1999) 3595 3602. Here, we used this optical imaging system to demonstrate that the backward digit span (DB) task activated the dorsolateral prefrontal cortex (DLPFC) of each hemisphere more than the forward digit span (DF) task in healthy adult volunteers, and higher performance of the DB task was closely related to the activation of the right DLPFC. These results suggest that visuospatial imagery is a useful strategy for the DB task. Optical tomography described here is a new modality of neuropsychological studies. PMID- 10808145 TI - Functional optimization of arithmetic processing in perfect performers. AB - Lesion and imaging studies to date have not clarified which sub-regions of the parietal lobe are specialized for arithmetic processing, and which perform supporting functions. We used functional magnetic resonance imaging to investigate parietal lobe function during arithmetic processing. Functional optimization was examined by analyzing regional differences in brain activation between perfect (100% accuracy) and imperfect performers. Perfect performers had significantly less activation only in the left angular gyrus, a finding that may be associated with skill mastery and long-term practice effects. The present results provide the first direct evidence of localized functional optimization for arithmetic processing in the human brain. PMID- 10808146 TI - Calcium channel blockers and angiotensin-converting enzyme inhibitors in the prevention of graft vasculopathy. PMID- 10808147 TI - Brain death and its impact on the donor heart-lessons from animal models. PMID- 10808148 TI - Prognostic value of serum cytokines in patients with congestive heart failure. AB - BACKGROUND: Increased levels of circulating cytokines have been previously reported in patients with congestive heart failure; however, whether they have prognostic implications is still unknown. The aim of this study was to assess the prognostic implications of elevated serum cytokines in patients with heart failure and to identify the predictors of cytokine activation. METHODS AND RESULTS: We assessed neurohormonal determinations, circulating cytokines, ejection fraction (EF) and end-diastolic and end-systolic left ventricular lengths in 87 patients (aged 57 +/- 9 years) with left ventricular dysfunction (EF 24% +/- 6%). In 48 patients, we also assessed cytokine receptors. During follow-up (mean, 14 +/- 9 months), 8 patients died and 12 had new heart failure episodes that required hospital admission, 5 of whom underwent heart transplantation. The univariate predictors of these events were serum interleukin 6 (IL-6) (p = 0.00001), New York Heart Association (NYHA) functional class (p = 0.0004), tumor necrosis factor-soluble receptor I (p = 0. 001), atrial natriuretic peptide (p = 0.002), tumor necrosis factor-soluble receptor II (p = 0.004), angiotensin II (p = 0.006), serum interleukin-1 beta (p = 0.01), and plasma renin activity (p = 0.02). Increased serum interleukin-6 (>10 pg/ml) was a significant predictor of death or new heart failure episodes according to the Kaplan-Meier survival method by log-rank test (p = 0.004). By Cox regression analysis, serum IL-6 (p = 0.0005) and the NYHA functional class (p = 0.005) were identified as independent predictors of prognosis. CONCLUSIONS: In patients with congestive heart failure, increased serum IL-6 was identified as a powerful independent predictor of the combined end point: death, new heart failure episodes, and need for heart transplantation. PMID- 10808149 TI - Invasive and non-invasive determinants of pulmonary hypertension in patients with chronic heart failure. AB - BACKGROUND: In patients with chronic heart failure, pulmonary hypertension is an important predictive marker of adverse outcome. Its invasive and non-invasive determinants have not been evaluated. OBJECTIVE: This study was performed to evaluate hemodynamic determinants of pulmonary hypertension in chronic heart failure and to compare the predictive value of Doppler indices with that of invasively measured hemodynamic indices. METHODS: Right heart catheterization and transthoracic echo-Doppler were simultaneously performed in 259 consecutive patients with chronic heart failure (ejection fraction 24% +/- 7%) who were in sinus rhythm and receiving optimized medical therapy. Systolic pulmonary artery pressure (sPAP), cardiac index, transpulmonary gradient pressure, and pulmonary wedge pressure (PWP) were measured invasively. Left atrial and ventricular systolic and diastolic volumes, the ratio of maximal early to late diastolic filling velocities (E/A ratio), deceleration time (DT) and atrial filling fraction (AFF) of transmitral flow, systolic fraction of forward pulmonary venous flow (SFpvf), and mitral regurgitation were quantified by echo-Doppler. RESULTS: Patients with pulmonary hypertension had greater left atrial systolic and diastolic dysfunction, more left ventricular diastolic abnormalities, and greater hemodynamic impairment. The correlations between systolic left ventricular indices, mitral regurgitation, and sPAP were generally poor. Among invasive and non-invasive measurements, PWP (r = 0.89, p < 0.0001) and SFpvf (r = -0.68, p < 0.0001) showed the strongest correlation with sPAP. When we compared all patients with those without mitral regurgitation, the correlations between E/A ratio (r = 0.56 vs r = 0. 74, p < 0.002), SFpvf (r = -0.68 vs r = -0.84, p < 0.03), and systolic pulmonary artery pressure were significantly stronger. Multivariate analysis revealed that PWP was the strongest invasive independent predictor of systolic pulmonary artery pressure in patients with (R(2) = 0.87, p < 0.0001) and without (R(2) = 0.90, p < 0.0001) mitral regurgitation. A PWP > or= 18 mm Hg (odds ratio [95% CL], 142 (41-570) was strongly associated with systolic pulmonary hypertension. Among non-invasive variables DT, SFpvf, and AFF were identified as independent predictors of sPAP in patients with (R(2) = 0.56, p < 0.0001) and without (R(2) = 0.78, p < 0.0001) mitral regurgitation. A DT < 130 (odds ratio [95% CL], 3.5 (1.3-8.5), SFfvp < 40% (odds ratio [95% CL], 333 (41 1,007), and AFF < 30% (odds ratio [95% CL], 2 (1.3-7) most strongly predicted systolic pulmonary hypertension. CONCLUSIONS: The results of this study indicate that in patients with chronic heart failure, venous pulmonary congestion is an important determinant of systolic pulmonary artery hypertension. Hemodynamic and Doppler determinants showed similar predictive power in identifying systolic pulmonary artery hypertension. PMID- 10808150 TI - Ten-year follow-up of critically ill patients undergoing heart transplantation. AB - BACKGROUND: The long-term result following heart transplantation appears very good despite complications of coronary atherosclerosis and cancer. Critically ill patients supported with mechanical devices remain a growing and difficult group in which long-term results need to be defined. The objective of this study was to review the 10-year follow-up of critically ill patients who underwent heart transplantation after support with mechanical devices. METHODS: We retrospectively analyzed all patients who underwent heart transplantation from 1986 to 1999 at the Montreal Heart Institute. RESULTS: Twenty-two patients (22/199, 11%) underwent heart transplantation after support with intra-aortic balloon pumps (n = 17) and total artificial hearts (n = 5). One hundred seventy seven patients (177/199, 89%) underwent heart transplantation without pre operative mechanical assistance. Patients with pre-operative mechanical assistance were younger (41 +/- 12 vs 48 +/- 10 years old, p = 0. 002), underwent a shorter waiting time to transplantation (2 +/- 2 vs 19 +/- 27 weeks, p = 0.004), and donor hearts had longer ischemic time (166 +/- 63 vs 137 +/- 49 minutes, p = 0.002) compared with patients without pre-operative mechanical assistance. One-month, 1-, 5-, and 10-year survival averaged 86% +/- 7%, 67% +/- 10%, 67% +/- 10%, and 59% +/- 12%, respectively, in patients with pre-operative mechanical assistance compared with 95% +/- 2%, 88% +/- 2%, 81% +/- 3%, and 74% +/- 4%, respectively, in patients without assistance, a significant difference (p = 0.04) that is mainly related to higher operative mortality in the former group. Although, we found no difference between the 2 groups in the 10-year freedom rate from acute rejection, infection, cancer, and coronary atherosclerosis, sepsis was the cause of 4 early deaths among patients with pre-operative mechanical assistance. CONCLUSION: Early and long-term survival was significantly decreased in critically ill patients with pre-operative mechanical assistance compared with other patients without pre-operative assistance. Sepsis is a dominant threat among patients who underwent heart transplantation with pre-operative mechanical assistance, and the lower survival is due mainly to the increased early mortality. PMID- 10808151 TI - Left ventricular function and mass after orthotopic heart transplantation: a comparison of cardiovascular magnetic resonance with echocardiography. AB - OBJECTIVE: We compared the assessment of left ventricular function and mass by M mode echocardiography (echo) with fast breath-hold cardiovascular magnetic resonance (CMR) in patients who received orthotopic heart transplantation. We also sought to establish the reproducibility of breath-hold CMR in this patient population. METHODS: We prospectively acquired 51 sets of echo and CMR data in 21 patients who had undergone orthotopic heart transplantation. We examined the intraobserver and interobserver reproducibility of breath-hold CMR in this group and compared it with published data. We compared the left ventricular ejection fraction (EF) and mass determined by echo with the CMR data. RESULTS: The average time between CMR and echo was 0 +/- 7 days (mean +/- SD), the time between each set of CMR-echo data acquisition was 5.1 +/- 4.1 months. Cardiovascular magnetic resonance showed good reproducibility in this population, with intraobserver percentage variability of 2.2% +/- 2.4% for EF and 3. 2% +/- 2.7% for mass, and interobserver percentage variability of 2. 4% +/- 1.9% for EF and 2.2% +/- 1.9% for mass. The Bland-Altman limits of agreement between echo and CMR were wide for both EF (-9. 6% to 15%) and mass, irrespective of the formula used (-61.3 to 198 g for the Bennett and Evans formula, -65.4 to 196.8 g for the American Society of Echocardiography (ASE) formula, -65.3 to 181 g for the Devereux formula, and 95.2 to 64.6 g for the Teichholz formula). CONCLUSION: Fast-acquisition CMR is reproducible in recipients of transplanted hearts. We found poor agreement with the results of echo. The choice of technique will depend on local resources as well as the clinical importance of the result. Echo remains readily available and gives rapid assessment of volumes, EF, and mass. However, the good reproducibility of CMR may make it a more suitable technique for long-term follow up of an individual or of a study population. PMID- 10808152 TI - Predictive factors and long-term evolution of early endothelial dysfunction after cardiac transplantation. AB - BACKGROUND: Abnormal coronary vasomotion appears to be a common finding after heart transplantation (HTx). However, the pathophysiology and outcome of this functional disturbance remains poorly understood. Aims of the study were to determine the prevalence, predictive factors and long-term evolution of endothelial dysfunction after cardiac transplantation. METHODS: The endothelium dependent coronary vasomotion of 50 patients, who showed angiographically normal coronary arteries, were studied early (at 3 +/- 1 months) and at follow-up (16 +/ 5 months) after HTx. Endothelial function was studied by selective infusion of serial doses of acetylcholine (ACh) (10(-8), 10(-7)and 10(-6) mol/l) in the left anterior descending coronary artery. Changes in mean luminal diameter after the infusion of each dose were evaluated by quantitative coronary angiography (QCA). RESULTS: At early study, 17 patients (34%) showed a vasoconstriction after maximal dose of ACh (-13.3 +/- 13%) indicative of endothelial dysfunction. Logistic regression analysis identified the following variables as independent predictors of early endothelial dysfunction: donor inotropic support (p = 0.004), female donor (p = 0.04) and rejection at the time of the study (p = 0.01). Forty one patients were re-studied at follow-up. Nine of them (22%) presented endothelial dysfunction. Early endothelial dysfunction was restored in 6 patients (43%) at follow-up. The number of episodes of rejection was the only variable associated to late endothelial dysfunction. CONCLUSIONS: Endothelial dysfunction is a common finding after cardiac transplantation. The pathogenesis of this functional disturbance appears to be donor-related and immune-mediated. The reversibility of this phenomenon observed at follow-up suggests the episodic nature of the immunologic injury. PMID- 10808153 TI - Changes in platelet activation associated with left ventricular assist system placement. AB - BACKGROUND: Thromboembolic and hemorrhagic complications are common in patients after left ventricular assist system (LVAS) placement. Platelet physiology may be involved in these complications. METHODS: Using flow cytometry, expression of CD62P and CD63 were analyzed as markers of platelet activation. Binding of annexin V was analyzed to determine platelet membrane asymmetry. Results from two patients who received a Novacor LVAS as a bridge to transplantation are reported. RESULTS: Patients' platelets showed increased CD62P and CD63 expression, yet annexin V binding was not increased. They also revealed suppression of thrombin activation following LVAS placement, which approached normal after transplantation. Heparin suppressed thrombin activation, whereas aspirin or dipyridamole did not. Suppression was attenuated by protamine sulfate and heparinase. CONCLUSIONS: Following LVAS placement, resting platelets demonstrate increased expression of activation markers. PMID- 10808154 TI - Long-term follow-up of pediatric cardiac transplant recipients on a steroid-free regime: the role of endomyocardial biopsy. AB - BACKGROUND: Endomyocardial biopsy is used in long-term follow-up of cardiac transplant recipients to detect sub-clinical rejection. The value of this for adults and children on triple-drug immunosuppression has been disputed. We investigated its value in children maintained on a steroid-free regime. METHODS: We used a retrospective review of annual surveillance biopsy results from children younger than 13 years at the time of cardiac transplant. RESULTS: In a series of 40 children older than 10 years, we found no evidence of rejection in 88/130 (67.7%) biopsies; 41/130 (31.5%) showed grade 1A rejection, and 1/130 (0.8%) showed grade 1B rejection. No grade 2, 3, or 4 biopsies were found. Nine patients with 1A rejection had subsequent grade 0 biopsies, without any adjustment in treatment. Seven children had treatment changes and repeat biopsies because of grade 1A biopsies. CONCLUSION: Significant late rejection is rare even in children on steroid-free maintenance. It is unlikely to be detected unexpectedly, and the practice of indefinite routine biopsy in children who are well is not justified. Future use should focus on individuals at higher risk of rejection. PMID- 10808155 TI - Does human leukocyte antigen matching influence the outcome of lung transplantation? An analysis of 3,549 lung transplantations. AB - BACKGROUND AND OBJECTIVE: Human leukocyte antigen (HLA) compatibility has been shown to improve the outcome of renal and cardiac transplantation. However, its impact on outcome following lung transplantation is not clear, with several single-center studies reporting inconsistent results. We studied the influence of HLA matching on survival and the development of rejection and obliterative bronchiolitis after lung transplantation, using data from the United Network for Organ Sharing/International Society for Heart and Lung Transplantation registry. METHODS: The study population included adult patients who received cadaveric lung transplants between October 1987 and June 1997 for whom HLA data were available. Two cohorts were examined, depending on the era of transplantation: (1) October 1987 to June 1997 (n = 3,549): Differences in actuarial survival as stratified by either the total number of HLA mismatches or the number of mismatches at each HLA locus were determined using a log-rank test. Multivariate logistic regression models were developed to determine independent predictors of survival at 1, 3, and 5 years following lung transplantation. (2) April 1994 to June 1997 (n = 1,796): The association of HLA mismatching with acute rejection and obliterative bronchiolitis was determined using a chi-squared analysis. RESULTS: Only 164 patients (4.6%) received lung grafts with 2 or fewer HLA mismatches. Univariate analyses demonstrated a significant difference in post-transplant survival by mismatch level, with the total number of HLA mismatches (p = 0.0008) and mismatching at the HLA-A locus (p = 0.002) associated with worse survival. Multivariate logistic regression demonstrated that the number of mismatches at the HLA-A and HLA-DR loci predicted 1-year mortality (incremental odds ratios 1.18, p = 0.01, and 1.15, p = 0. 03, respectively). The total number of HLA mismatches predicted 3- and 5-year mortality (incremental odds ratios 1.13 at 3 years, p = 0. 0004, and 1.14 at 5 years, p = 0.0002). However, other covariates such as repeat transplantation, transplantation for congenital heart disease, advanced recipient age, and an early era of transplantation were stronger predictors of mortality. We found no significant association between HLA mismatching and the development of obliterative bronchiolitis, although there was an association between mismatching at the HLA-A locus and acute rejection episodes requiring hospital admission (p = 0.008). We also found no association between mismatching at the HLA-B locus and rejection episodes requiring either hospitalization or the alteration of anti-rejection medications (p = 0.034). CONCLUSION: Although the number of HLA mismatches at the HLA-A and HLA-DR loci predicted 1-year mortality and the total number of mismatches predicted 3- and 5 year mortality following lung transplantation, the effect of each covariate was small in this multicenter study of 3,549 patients. Further close follow-up of registry patients is necessary to determine the effect of HLA matching on long term survival and freedom from obliterative bronchiolitis and rejection following lung transplantation. A prospective study of HLA matching for lung transplantation should not yet be considered in view of the small number of grafts with 2 or fewer mismatches and the modest effect of HLA matching on outcome. PMID- 10808156 TI - Lung transplantation following lung volume reduction surgery. AB - OBJECTIVE: Lung volume reduction surgery (LVRS) has been proposed as a possible alternative treatment to lung transplantation (LTX) for selected patients with end-stage emphysema. But whether LVRS is a temporary or permanent alternative to LTX is still under investigation. The aim of this study was to analyze the course of patients undergoing LVRS followed by subsequent LTX. METHODS: Fifteen patients (10 male, 5 female, mean age 53.3 +/- 1.7 years) out of 102 patients, who underwent LVRS between September 1994 and August 1998, underwent LTX 19.6 +/- 3.1 months after LVRS (range 1.7 to 37.6 months) between June 1996 and October 1998. In 9 patients bilateral LVRS was performed, in 6 patients unilateral LVRS. Subsequent LTX was performed bilaterally in 10 patients and unilaterally in 5 patients (1 of these on the contralateral side) to the previous LVRS. The course of lung function and clinical outcome were analyzed in these 15 patients. RESULTS: Mean forced expiratory volume in 1 second (FEV(1)) in the 15 patients prior to LVRS was 18.3 +/- 1.2% of predicted (%p) and increased to 27.0 +/- 2.9 %p (best value within the first 6 months postLVRS) (p = 0.043). In 8 of these patients (non-responders) (53%) LVRS failed to improve FEV(1), whereas in the other 7 patients (responders) (47%) a significant improvement was detected (FEV(1) 18.1 +/- 1.8 %p and 31.9 +/- 3.7 %p, pre- and post-LVRS, respectively, p = 0.003), but declined after 6 to 36 months. At the time of listing for LTX the mean FEV(1) was 18.0 +/- 1.9 %p (no difference between the 2 groups). LTX was performed 15.5 +/- 3.6 months (non-responders) and 25.7 +/- 4.6 months (responders) after LVRS. FEV(1) improved to 81.0 +/- 5.6 %p after LTX (p < 0.001 compared to pre-LTX). The mortality after LVRS was 0%. The 3-month mortality after LTX was 20% (1 patient with primary organ failure, 1 patient with ongoing rejection, 1 patient with sepsis). All 3 patients belonged to the group of nonresponders. Two patients died 5. 5 and 8.5 months after LTX (13.3%) due to fungal infection (Aspergillus spp.) and MRSA sepsis, respectively (1 non responder, 1 responder). CONCLUSIONS: Successful LVRS delays the need for LTX and offers better conditions for LTX. However, patients without functional improvement after LVRS have a high perioperative risk at subsequent LTX. PMID- 10808157 TI - Obstruction of the inferior vena cava following total heart lung transplantation: successful treatment by balloon angioplasty. AB - We report a case of inferior vena caval obstruction early after total heart-lung transplantation. The patient presented with edema of both legs and a venous hum at the base of the right lung. This complication was successfully treated by balloon angioplasty, avoiding the need for re-operation. PMID- 10808158 TI - Anti-CD20 monoclonal antibody therapy in Epstein-Barr Virus-associated B cell lymphoma following lung transplantation. AB - Post-transplant lymphoproliferative disease is a complication of bone marrow and solid organ transplantation, mostly associated with Epstein-Barr virus infection and chronic immunosuppression. Even if spontaneous resolution after cessation of immunosuppressive therapy can be observed, the prognosis of this disorder is usually poor with a low response to specific treatment. We describe a case of B cell lymphoma of the nasopharynx occurring 6 months after double-lung transplantation. In spite of its monoclonal nature, anti-CD 20 monoclonal antibody given in the presence of reduced immunosuppression resulted in a complete response. The patient also received "consolidation" radiation therapy to prevent the recurrence. The treatment was well tolerated with minimal side effects. The patient was asymptomatic and had a well functioning graft more than 1 year after therapy. PMID- 10808160 TI - Blood loss and transfusion requirements in patients implanted with a mechanical circulatory support device undergoing cardiac transplantation. AB - Patients implanted with mechanical circulatory support devices (MCSD's) are at high risk for post-operative bleeding at cardiac transplantation. However, the magnitude of the risk and transfusion requirements for MCSD patients at the time of transplantation have not been previously reported. The purpose of this study was to characterize and compare the bleeding characteristics and transfusion requirements of 3 sub-groups of cardiac transplant patients: primary (n = 45), redo (n = 26), and MCSD (n = 23) patients. PMID- 10808159 TI - The use of donor hearts with left ventricular hypertrophy. AB - BACKGROUND: We reviewed 37 patients who received donor hearts with left ventricular hypertrophy (LVH) to determine which factors affected outcomes. METHODS: Thirty-seven patients underwent orthotopic heart transplantation (1994 through 1998) with donor hearts qualified as having LVH by echocardiography (EC) and/or electrocardiogram (ECG). We performed univariate analysis on 18 donor and recipient risk factors for mortality. We calculated 12-month survival curves using Kaplan-Meier estimates and compared them using the log-rank test. A contemporaneous cohort of 221 patients who received optimal hearts within the same institution served as a control for survival. RESULTS: Median follow-up was 18 months (1 to 53). Median recipient age was 58 ye ars (25 to 75), and median donor age was 47 years (12 to 63). Median donor/recipient height and weight ratios were 1.01 (0.9 to 1.19) and 1.16 (0.77 to 2.02), respectively. Two-month survival was 86.4%, and 12-month survival was 73.0%. Survival for the control group was 91. 6% at 2 months and 86.9% at 12 months. Clinically inferior survival curves were observed when donors had known hypertension (n = 17, 95% vs 71% at 2 months, 76% vs 65% at 12 months), ischemia > 180 minutes (n = 18, 95% vs 72% at 2 months, 78% vs 65% at 12 months), LVH by ECG (n = 10, 85% vs 80% at 2 months, 77% vs 56% at 12 months), and greater than mild or unknown ECHO grade (n = 18, 89% vs 72% at 2 months, 84% vs 59% at 12 months, p = 0.11). CONCLUSIONS: Donor hearts with mild LVH may be used selectively, particularly if there are no ECG criteria and if ischemia time is short. Caution is indicated for donors with documented history of hypertension. Precise measurement of LV wall thickness by EC is needed in all donors to estimate severity and to complement ECG interpretation. PMID- 10808161 TI - Lack of acute cyclosporine nephrotoxicity in late heart-transplant recipients. AB - BACKGROUND: Cyclosporine induces daily renal hypoperfusion in subjects with normal atrial natriuretic peptide (ANP) levels, but its acute effects in heart transplant patients with increased ANP remain to be determined. METHODS: Cyclosporinemia and creatinine clearance were monitored during 7 hours following cyclosporine administration in 6 heart transplant patients. RESULTS: CONCLUSIONS: No acute cyclosporine-induced decrease in creatinine clearance was observed after heart transplantation. These data suggest that maintenance cyclosporine dose may be less nephrotoxic than suspected and that increased ANP might protect the renal function late after heart transplantation. PMID- 10808163 TI - The diverse potential effector and immunoregulatory roles of mast cells in allergic disease. AB - Mast cells are of hematopoietic origin but typically complete their maturation in peripheral connective tissues, especially those near epithelial surfaces. Mast cells express receptors that bind IgE antibodies with high affinity (FcepsilonRI), and aggregation of these FcepsilonRI by the reaction of cell-bound IgE with specific antigens induces mast cells to secrete a broad spectrum of biologically active preformed or lipid mediators, as well as many cytokines. Mast cells are widely thought to be essential for the expression of acute allergic reactions, but the importance of mast cells in late-phase reactions and chronic allergic inflammation has remained controversial. Although it is clear that many cell types may be involved in the expression of late-phase reactions and chronic allergic inflammation, studies in genetically mast cell-deficient and congenic normal mice indicate that mast cells may be critical for the full expression of certain features of late-phase reactions and may also contribute importantly to clinically relevant aspects of chronic allergic inflammation. Moreover, the pattern of cytokines that can be produced by mast cell populations, and the enhancement of such cytokine production in mast cells that have undergone IgE dependent up-regulation of their surface expression of FcepsilonRI, suggests that mast cells may contribute to allergic diseases (and host defense) by acting as immunoregulatory cells, as well as by providing effector cell function. PMID- 10808164 TI - Atopic dermatitis: new insights and opportunities for therapeutic intervention. AB - Atopic dermatitis (AD) is a chronic inflammatory skin disease that frequently predates the development of allergic rhinitis or asthma. It is an important skin condition with significant costs and morbidity to patients and their families; the disease affects more than 10% of children. Recent studies have demonstrated the complex interrelationship of genetic, environmental, skin barrier, pharmacologic, psychologic, and immunologic factors that contribute to the development and severity of AD. The current review will examine the cellular and molecular mechanisms that contribute to AD as well as the immunologic triggers involved in its pathogenesis. These insights provide new opportunities for therapeutic intervention in this common skin condition. PMID- 10808165 TI - Cytokine receptor signaling pathways. AB - Cytokines represent a diverse group of molecules that transmit intercellular signals. These signals may either be autocrine (where the same cell both produces the cytokine and responds to it) or paracrine (where the cytokine is made by one cell and acts on another). Both these situations can occur simultaneously. Cytokines use multiple signaling pathways. This review will focus on signaling by type I cytokines and in particular on signaling by the IL-2 family of cytokines, as an illustrative example. The major signaling pathway that will be discussed is the Jak-STAT pathway, although other pathways will also be reviewed. The Jak-STAT pathway is a very rapid cytosol-to-nuclear signaling pathway that underscores how quickly extracellular signals can be transmitted to the nucleus. Aspects related to cytokine redundancy, pleiotropy, and specificity will be discussed. PMID- 10808166 TI - Do allergies protect against the effects of a rhinovirus cold? PMID- 10808167 TI - Chemokines and atopic dermatitis. PMID- 10808168 TI - Increased nitric oxide production in the respiratory tract in asymptomatic pacific islanders: an association with skin prick reactivity to house dust mite. AB - BACKGROUND: Exhaled nitric oxide (NO) is increased in asthma and may also be increased in subclinical airway inflammation. The relationship between atopy and subclinical airway inflammation in the pathogenesis of asthma remains unclear. We have evaluated the relationship between exhaled NO levels and skin prick test reactivity to 8 common allergens in 64 asymptomatic adult Pacific Islanders. Pacific Islanders were studied as a racial group with major morbidity from asthma. OBJECTIVE: Our purpose was to determine whether asymptomatic subjects with skin prick test reactivity to common allergens have elevated NO levels. METHODS: All subjects underwent full lung function testing and skin prick testing. Exhaled and nasal NO levels were measured by chemiluminescence (Logan LR2000 analyzer) with use of the single-breath and breath-holding techniques, respectively. RESULTS: House dust mite (HDM) reactivity was seen in 38 of 64 (56%). Exhaled NO levels (median 8.9 ppb, range 2.9-47.3 ppb) and nasal NO levels (527.5 +/- 181.5 ppb) lay above the normal European range in 30% and 25% of subjects, respectively. HDM reactivity was associated with higher exhaled NO levels (P <. 0005) and higher nasal NO levels (P =.01). In HDM-sensitive subjects the wheal size for HDM correlated with exhaled NO levels (r = 0.35, P =.04) and nasal NO levels (r = 0.40, P =.01). On multivariate analysis, exhaled NO levels were independently and positively related to the severity of HDM reactivity (P =.01) and nasal NO levels (P <.02), equation R(2) = 0.27. CONCLUSION: NO levels are elevated in a significant proportion of asymptomatic Pacific Islanders and are associated with HDM sensitivity. This may denote subclinical airway inflammation in this population and suggests that exposure to HDM in atopic individuals might play an important role in the early pathogenesis of asthma. PMID- 10808169 TI - Sputum matrix metalloproteinase-9: tissue inhibitor of metalloproteinase-1 ratio in acute asthma. AB - BACKGROUND: The ratio of matrix metalloproteinase-9 (MMP-9) and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1) may be a marker of the balance between airway tissue destruction and repair. TIMP-1 may potentially contribute to the pathogenesis of increased submucosal extracellular matrix deposition in asthma. OBJECTIVE: Our purpose was to assess the variation in sputum MMP-9 and TIMP-1 during acute asthma. METHODS: We evaluated the MMP-9 and TIMP-1 balance in sputa of 16 asthmatic patients admitted with spontaneous exacerbation, conducting measurement before (day 1) and after methylprednisolone infusion therapy (days 2, 3, 5, and 7), and on remission days. RESULTS: Peak expiratory flow and eosinophilic cationic protein levels were significantly (P <.05) improved within 7 days in all patients. Sputum MMP-9 levels on day 2 tended to be lower than on day 1, but not significantly. Zymography revealed that the main enzyme was identified immunologically as MMP-9, and gelatinase activity on day 1 had a tendency to decrease for the following 7 days. The TIMP-1 levels gradually increased until day 5, were significantly (P <.05) high on day 5, and decreased on day 7. The MMP-9/TIMP-1 molar ratios were significantly (P <.05) decreased on days 2, 3, 5, and 7 compared with day 1. Sputum levels of MMP-9 and TIMP-1 and the MMP-9/TIMP-1 molar ratios on day 1 were significantly higher (P <.02) than those on remission days. CONCLUSIONS: An imbalance between MMP-9 and TIMP-1 was present in acute asthma, with an excess of MMP-9 resulting in a high ratio of MMP 9/TIMP-1 before treatment, and over time with glucocorticosteroid the TIMP-1 levels rose, dropping the ratio of MMP-9/TIMP-1. It was suggested that overproduction of MMP-9 and TIMP-1 after asthma exacerbation might contribute significantly to airway tissue remodeling and that TIMP-1 production in acute asthma might not be suppressed by glucocorticosteroid. PMID- 10808170 TI - Mometasone furoate antagonizes AMP-induced bronchoconstriction in patients with mild asthma. AB - BACKGROUND: Mometasone furoate (MF) is a new potent corticosteroid for use in treating asthma. OBJECTIVE: To test the lower range of the dose-response curve, effects of MF delivered by dry powder inhaler (DPI) on AMP-induced bronchoconstriction were compared with those of placebo. METHODS: In a placebo controlled, 3-phase cross-over, single-center, double-blind study, 15 patients with mild asthma were randomized to three 2-week treatment phases (separated by 4 week washout phases) with MF DPI 50 microg twice daily, MF DPI 100 microg twice daily, or placebo. AMP challenge was performed before and at the end of each treatment phase. RESULTS: Thirteen patients completed all 3 phases and were included in the primary efficacy analysis. Treatment with MF DPI 50 microg twice daily or with MF DPI 100 microg twice daily significantly reduced the bronchoconstrictor response to AMP, displacing the dose-response curve to the right by 2.81 and 3.11 doubling dilutions, respectively, compared with placebo (P <.001). The improvement in FEV(1) over the 2-week treatment phase was significantly (P < or =.033) greater during treatment with MF DPI 50 microg or 100 microg twice daily than with placebo. Peak expiratory flow rate, wheezing scores, difficulty breathing scores, nocturnal awakenings requiring salbutamol, and puffs of salbutamol per day also indicated a greater improvement in respiratory function and symptoms of asthma with MF DPI 50 or 100 microg twice daily than with placebo. Both doses of MF DPI were well tolerated. CONCLUSIONS: Treatment with low doses of MF DPI decreased airway responsiveness to AMP challenge and improved secondary measures of pulmonary function and asthma symptoms. PMID- 10808171 TI - Risk of cataract among users of intranasal corticosteroids. AB - BACKGROUND: Oral corticosteroid users are at increased risk of cataract, but the risk among intranasal corticosteroids users is unknown. OBJECTIVE: Our purpose was to describe the risk of cataract among users of intranasal steroids. METHODS: A retrospective observational cohort study of cataract incidence was conducted among users of oral and intranasal steroids identified from the United Kingdom based General Practice Research Database with a nested case-control analysis to control for confounding factors. The study population included 286,078 subjects aged less than 70 years old drawn from 350 general practices in England and Wales. Patients were classified as users of only intranasal corticosteroids, users of only oral corticosteroids, and nonusers of either medication. Computerized medical records were used to identify cases of cataract. Two hundred twenty-five cases were randomly selected for validation against general practitioners' held referral and hospitalization letters. RESULTS: The incidence rate of cataract (1.0/1000 person-years) among users of intranasal corticosteroids was similar to the incidence rate among nonusers. However, oral corticosteroid users were at higher risk of cataract (2.2/1000 person-years). Approximately 70% of intranasal corticosteroid exposure was to beclomethasone dipropionate only; the event rate in this group was similar to that in the unexposed group. Cataract risk did not increase with the number of prior prescriptions for intranasal corticosteroids. CONCLUSION: The use of intranasal corticosteroids was not associated with an increased risk of cataracts in this study population. PMID- 10808172 TI - Concomitant montelukast and loratadine as treatment for seasonal allergic rhinitis: a randomized, placebo-controlled clinical trial. AB - BACKGROUND: Nasal challenge studies have suggested histamine and cysteinyl leukotrienes are important proinflammatory mediators in allergic rhinitis. This study was designed to determine the efficacy of montelukast, a cysteinyl leukotriene receptor antagonist, administered alone or concomitantly with loratadine, an H(1)-receptor antagonist, in seasonal allergic rhinitis. OBJECTIVE: The purpose of this study was to determine the effect of concomitant use of montelukast and loratadine in the treatment of seasonal allergic rhinitis. METHODS: In this multicenter (N = 12) double-blind, randomized, parallel-group, placebo-controlled 2-week trial, 460 men and women, aged 15 to 75 years, with spring seasonal allergic rhinitis were randomly allocated to receive 1 of the following 5 treatments: montelukast 10 or 20 mg, loratadine 10 mg, montelukast 10 mg with loratadine 10 mg, or placebo, once daily in the evening. The primary end point was daytime nasal symptoms score (average of congestion, rhinorrhea, itching, and sneezing). Other end points were eye symptoms, nighttime symptoms, individual daytime nasal symptoms, global evaluations (patient's and physician's), and rhinoconjunctivitis quality-of-life scores. RESULTS: Concomitant montelukast with loratadine improved the primary end point significantly (P <.001) compared with placebo and each agent alone. Compared with placebo, montelukast with loratadine also significantly improved eye symptoms, nighttime symptoms, individual daytime nasal symptoms, global evaluations, and quality of life. Montelukast alone and loratadine alone caused modest improvements in rhinitis end points. All treatments were similarly well tolerated. CONCLUSIONS: Concomitant montelukast with loratadine provided effective treatment for seasonal allergic rhinitis and associated eye symptoms with a safety profile comparable with placebo. PMID- 10808173 TI - Effects of allergic inflammation of the nasal mucosa on the severity of rhinovirus 16 cold. AB - BACKGROUND: Despite the strong association of asthma exacerbations with rhinovirus (RV) infection, inoculation of asthmatic subjects with RV only causes small changes in lower airway function, suggesting that RV infection is not itself sufficient to provoke asthma exacerbations. OBJECTIVE: Our purpose was to test whether allergic inflammation increases the airway response to RV infection. METHODS: We compared the severity of RV type 16-induced colds in 2 groups of 10 subjects with allergic rhinitis. One group received 3 nasal challenges with allergen and the other received challenges with placebo over the week before nasal inoculation with RV type 16 (4000 tissue culture infective dose 50% per subject). Subjects kept symptom diaries and were assessed with spirometry, methacholine challenge, nasal lavage, and sputum induction on days 2, 4, 7, 10, 15, and 30 after inoculation. RESULTS: The 2 groups developed equal rates of infection (90%), similar cold symptoms (Jackson score median [interquartile range], 11 [6-33] vs 20.5 [6-42] for allergen and placebo groups respectively, P =.54), and similar changes in cellular profile and in IL-6 and IL-8 concentrations in nasal lavage fluid and induced sputum after RV inoculation. The incubation period was significantly longer in the allergen group (2.5 [1-5.5] vs 1 [1-1] day, P =.03) and the duration of cold symptoms was shorter (5 [4-7] vs 8.5 [6-10] days, P =.008). We also found an inverse correlation between the percent of eosinophils in nasal lavage fluid before inoculation and the severity of cold symptoms (r = -0.58, P =. 008). CONCLUSION: In subjects with allergic rhinitis, augmented nasal allergic inflammation before inoculation with RV type 16 does not worsen the severity of cold symptoms but delays their onset and shortens their duration. PMID- 10808174 TI - Children at risk for asthma: home allergen levels, lymphocyte proliferation, and wheeze. AB - BACKGROUND: Allergic asthma is a common childhood disease. Although T-lymphocyte activation plays a critical role in allergic asthma, the environmental factors promoting lymphocyte activation in children are not well defined. OBJECTIVE: In a cohort of children at risk for asthma (n = 114), we determined whether the levels of cockroach (Bla g 1 or 2), house dust mite (Der f 1), and cat allergen (Fel d 1) in the home during infancy was associated with subsequent allergen-specific lymphocyte proliferation in later life. METHODS: Dust samples from multiple sites in the home were collected at 3 months of age and were measured for allergen levels. Serial questionnaires were applied. At a median age of 2 years, PBMCs were isolated and lymphocyte proliferation to the home allergens and PHA was determined. RESULTS: Increased lymphocyte proliferative responses to Bla g 2 were associated with higher home levels of Bla g 1 or 2 (P for trend with kitchen Bla g levels =.011), in analyses adjusting for cold in the past week. Proliferative responses to Der f 1 were higher in homes with family room levels of Der f 1 > or =10 microg/g dust than in homes with Der f 1 <2 microg/g, but differences were not significant in analyses adjusting for cold (P =. 15). Repeated wheeze in the first 2 years of life was associated with increased allergen-specific and PHA proliferative responses. CONCLUSION: Early-life cockroach allergen exposure at 3 months of age predicts allergen-specific lymphocyte proliferative responses at a median of 2 years of age. PMID- 10808175 TI - Glucocorticoid resistance in asthma is associated with elevated in vivo expression of the glucocorticoid receptor beta-isoform. AB - BACKGROUND: Glucocorticoid-resistant bronchial asthma is characterized by failure of corticosteroids to suppress key asthma-relevant, cell-mediated inflammatory responses in the airways. OBJECTIVE: The mechanism of this phenomenon is not clear but may involve aberrant expression of the beta-isoform of the glucocorticoid receptor. METHODS: We have measured expression of the alpha- and beta-glucocorticoid receptor isoforms in tuberculin-driven cutaneous cell mediated inflammatory lesions in people with asthma who are glucocorticoid sensitive and resistant after 9 days of therapy with oral prednisolone (40 mg/day) or matching placebo in a random order, crossover design. RESULTS: After placebo therapy, the mean numbers of cells expressing glucocorticoid receptor alpha immunoreactivity in the lesions evoked in glucocorticoid-sensitive and resistant patients with asthma were statistically equivalent. The numbers of cells expressing glucocorticoid receptor beta were significantly elevated in the patients who were glucocorticoid resistant, resulting in an 8-fold higher ratio of expression of glucocorticoid receptor alpha/glucocorticoid receptor beta in the patients who were glucocorticoid sensitive. Glucocorticoid receptor alpha/glucocorticoid receptors beta were colocalized to the same cells. Oral prednisolone therapy was associated with a significant decrease in the numbers of cells expressing glucocorticoid receptor alpha but not glucocorticoid receptor beta in the subjects who were glucocorticoid sensitive. No significant change was found in the numbers of cells expressing glucocorticoid receptor alpha and glucocorticoid receptor beta in the patients who were glucocorticoid resistant. Prednisolone therapy reduced the ratio of glucocorticoid receptor alpha/glucocorticoid receptor beta expression for the patients who were glucocorticoid sensitive to a level seen in the patients who were glucocorticoid resistant before therapy. CONCLUSION: Because glucocorticoid receptor beta inhibits alpha-glucocorticoid receptor-mediated transactivation of target genes, the increased expression of glucocorticoid receptor beta in inflammatory cells might be a critical mechanism for conferring glucocorticoid resistance. PMID- 10808176 TI - Fetal and neonatal IL-13 production during pregnancy and at birth and subsequent development of atopic symptoms. AB - BACKGROUND: Cytokine production at the materno-fetal interface may influence the development of atopy-predisposing immune responses. Because IL-13 possesses IL-4 like activity and may regulate the immune responses observed in atopy, it may contribute to the expression of the atopic phenotype initiated during intrauterine life. OBJECTIVE: We sought to examine IL-13 expression by fetal and neonatal cells and the placenta. METHODS: The production of IL-13 by neonatal and fetal T cells was examined by culturing the cells in the presence or absence of PHA. Production of IL-13 at term was considered in the context of the later development of atopic disease in the child. IL-13 expression in the placenta was assessed by using immunohistochemistry. RESULTS: IL-13 immunoreactivity within the placenta was restricted to 16 to 27 weeks' gestation (6/6 positive vs 0/10 at >27 weeks' gestation). In contrast, spontaneous release of IL-13 by fetal mononuclear cells was first observed from 27 weeks' gestation but was undetectable after 37 weeks' gestation. PHA-stimulated mononuclear cells showed increased IL-13 levels in 80% of samples. Term babies (>37 weeks' gestation) with a parental history of atopy with atopic symptoms by 3 years of age produced significantly lower concentrations of PHA-induced IL-13 when compared with babies with no parental history of atopy (P =.034). CONCLUSION: Thus babies at risk of atopic disease in infancy display defective IL-13 production at birth. This may represent an inherent immaturity in the development of T cell-cytokine responses in babies at genetic risk for atopy or could be a consequence of downregulation of responses by other factors. Normal pregnancy, irrespective of atopic status, is associated with the production of appreciable quantities of IL-13 initially by the placenta and subsequently by the fetus. The regulation of this production and its consequences for the mother and fetus remains to be elaborated. PMID- 10808177 TI - HLA-DRB1*01 alleles are associated with sensitization to cockroach allergens. AB - BACKGROUND: Sensitization to cockroach allergens is an important epidemiologic risk factor for asthma, particularly among African Americans living in urban environments. A recent genome screen in the Hutterites, a white founder population, identified a linkage between an HLA-linked marker and sensitization to cockroach allergens. OBJECTIVE: Our purpose was to determine whether alleles at one or more HLA loci are associated with sensitization to cockroach allergens in ethnically diverse populations. METHODS: Alleles at 14 HLA region loci were studied in the Hutterites. On the basis of these results, selected loci were examined in 54 African Americans with cockroach sensitization (cases) and 65 African Americans without cockroach sensitization (controls). Sensitivity to cockroach allergens was assessed in both samples by skin prick test to purified cockroach allergens (Periplaneta americana and Blatella germanica). RESULTS: Significant associations between cockroach allergies and DRB1*0101 (P(corrected) =.0066), DQA1*0101 (P(corrected) =.0012), and DQB1*0501 (P(corrected) =.00096) were detected in the Hutterites. In the African American sample, the most significant association was with the DRB1*0102 allele (P(corrected) =.0088, odds ratio 16.4, 95% confidence interval 2.0, 131). The DRB1*0101 allele was infrequent in the African American sample (frequency 0.06) and the DRB1*0102 allele was absent in the Hutterites. DRB1*0101 and DRB1*0102 are closely related alleles that differ from nearly all other DRB1 alleles at 3 amino acids in the 1 peptide binding domain of the HLA-DR molecule. CONCLUSIONS: The DRB1*0101 allele in the Hutterites and the DRB1*0102 allele in African Americans confer risk for cockroach sensitization. Elucidating this interaction at the molecular level may allow for more targeted treatment and prevention of atopic asthma in inner-city populations. PMID- 10808178 TI - Deficient cytokine response of human allergen-specific T lymphocytes from humanized SCID mice and reconstitution by professional antigen-presenting cells. AB - BACKGROUND: Hu-PBL-SCID mice generated by the transfer of PBMCs from atopic individuals may provide a physiologic in vivo model for investigating human responses to allergens and potential approaches toward immunotherapy. OBJECTIVE: This study was undertaken to investigate the functional activity and cytokine profile of human allergen-reactive T lymphocytes isolated from hu-PBL-SCID mice. METHODS: PBMCs from allergic individuals were coinjected with allergen into SCID mice. Human lymphocyte migration and phenotype were established by reverse transcription-PCR and immunohistochemistry, IgE levels in sera were determined, and the frequency of allergen-reactive cytokine-producing T lymphocytes was established. RESULTS: After immunization with allergen, specific IgE levels in hu PBL-SCID sera were comparable with levels in donor sera. Although the majority of lymphocytes remained in the peritoneum, significant numbers of T lymphocytes were located in the spleen, where human IL-4, IL-5, and IFN-gamma messenger RNA expression was detected after stimulation with PHA and phorbol myristate acetate. Failure to induce cytokine production by human T lymphocytes isolated from the peritoneum and spleen of hu-PBL-SCID mice by allergen was reversed by stimulating with allergen in the presence of exogenously added IL-2 and antigen-presenting cells (APC), particularly CD14(+) monocytes. Under these conditions, allergen reactive T cells expressed a T(H)2-like phenotype. CONCLUSIONS: These data suggest that, after initial activation and induction of antibody production, human T lymphocytes enter a state of unresponsiveness, arising from a loss of human professional APC, in hu-PBL-SCID mice. The use of hu-PBL-SCID mouse models in studies on therapeutic approaches for allergy may benefit from the additional transfer of human professional APC. PMID- 10808179 TI - gp120- and TNF-alpha-induced modulation of human B cell function: proliferation, cyclic AMP generation, Ig production, and B-cell receptor expression. AB - BACKGROUND: It is well known that HIV-1 infection induces profound alterations in the immune system, including hyperactivation of B cells. TNF-alpha induces HIV-1 replication and immunodysregulation, including polyclonal B-cell activation. OBJECTIVE: We sought to determine the effects of surface-binding HIV-1 envelope glycoprotein (gp120) and TNF-alpha on human B-cell function. METHODS: HIV-1 seronegative peripheral blood human B cells were purified and activated by CD40 mAb and IL-4. In vitro studies of B-cell proliferation, cyclic AMP (cAMP) generation, receptor expression, and Ig production were performed. RESULTS: gp120, an Ig superantigen, stimulated HIV-1 seronegative and HIV-1 seropositive human B-cell cAMP generation, proliferation, and Ig production. These gp120 induced B-cell responses were demonstrated to be specific as evidenced by the abrogation of the stimulatory response in the presence of anti-gp120 mAb, blocking of CD4 resulting in no change on gp120-induced B-cell responses, and the binding of gp120 in these B cells. TNF-alpha also stimulated cAMP generation, proliferation, and Ig production in B cells, and the binding of gp120 to these B cells stimulated by TNF-alpha further enhanced cell proliferation, cAMP generation, and Ig production. Antigenic expression of the B-cell receptor CD79b was down-regulated by gp120 but was not altered by the addition of TNF-alpha. CONCLUSION: gp120 modulation of TNF-alpha-induced B-cell receptor- and cAMP mediated signal transduction events may be involved in the B-cell abnormalities observed in HIV-1 infection. PMID- 10808180 TI - DiGeorge anomaly: a comparative study of the clinical and immunologic characteristics of patients positive and negative by fluorescence in situ hybridization. AB - BACKGROUND: DiGeorge anomaly (DGA) is defined as a field defect characterized by dysmorphic facies, hypoparathyroidism, congenital heart defects, and a deficiency in cell-mediated immunity, usually associated with a microdeletion in chromosome 22q11.2. Data correlating clinical and genetic information, especially in terms of the extent of the immunodeficiency and infectious complications, are scant. OBJECTIVE: The goal of this study was to define the severity of the immunodeficiency and infectious illnesses in DGA patients with characteristic clinical and genetic findings and compare them with a similar group of patients without a microdeletion in chromosome 22q11.2. METHODS: A retrospective chart review of patients referred for evaluation of DGA to our immunology service from 1989 to 1995 was conducted. Clinical and immunologic data were collected from their initial evaluation. Patients meeting at least 3 of 4 of these criteria were considered to meet strict clinical diagnostic criteria for DGA, and the results of analysis for a microdeletion in chromosome 22q11.2 for each patient was noted. RESULTS: Sixteen of the 22 patients meeting strict clinical criteria for DGA were available for analysis for the microdeletion at chromosome 22q.11.2. Of these, 13 (81%) were positive by fluorescence in situ hybridization (FISH); 9 of 13 (69%) had low CD3 numbers, 6 of 10 assayed (60%) had low thymulin levels; 10 of 13 (77%) had low CD4 numbers, and 10 of 12 (83%) had absent or small thymus glands. B cells were increased in 9 of 13 (69%) patients. Mitogen and antigen responses were normal in 6 of 7 (86%) patients tested. Eight of 13 (62%) had a history of increased frequency of infectious illnesses. All had recurrent respiratory infections, including sinusitis, otitis media, and pneumonia. Three of the 16 patients tested (19%) were FISH negative. Two of 3 (67%) had low CD3 and CD4 numbers. B cells were elevated in all patients. All had recurrent respiratory infections, low thymulin levels, and absent thymus glands. CONCLUSIONS: Contrary to traditional descriptions, this group of clinically and genetically defined patients with DGA had a predominantly mild cell-mediated immunodeficiency syndrome usually associated with infections characteristic of humoral immunodeficiencies. The patients who were FISH positive did not differ significantly from those that were FISH negative in terms of clinical and immunologic findings or infectious complications. PMID- 10808181 TI - Comparison of allergen-stimulated dendritic cells from atopic and nonatopic donors dissecting their effect on autologous naive and memory T helper cells of such donors. AB - BACKGROUND: Because of their production of IL-12, mature dendritic cells (DC) are potent inducers of T(H)1 responses. However, recent reports have demonstrated that DCs can also induce T(H)2 differentiation. OBJECTIVE: In the current study we investigated which immune response is induced by DCs in naive CD45RA(+) or memory CD45R0(+) CD4(+) T cells from atopic individuals (patients with grass pollen, birch pollen, or house dust mite allergy) compared with nonatopic control subjects. METHODS: Immature DCs, generated from peripheral blood monocytes from atopic and nonatopic donors, were pulsed with the respective allergen and fully matured. Then the mature DCs were cocultured in vitro with autologous naive (CD45RA(+)) and memory (CD45R0(+)) CD4(+) T cells and cytokine and IgE production were measured by ELISA. RESULTS: After the second restimulation with allergen pulsed DCs, naive as well as memory autologous CD4(+) T cells from atopic but not from nonatopic donors showed an enhanced production of the T(H)2-type cytokines IL-4, IL-5, and IL-10, resulting in an increased IgE production, whereas IFN gamma production and proliferation were not different. IL-12 production and surface marker expression of DCs derived from atopic and nonatopic donors did not differ and addition of neutralizing anti-IL-12 mAbs did not increase IL-4 but diminished IFN-gamma production. CONCLUSION: These data indicate that mature DCs are able to induce naive and activate allergen-specific T helper cells to produce T(H)2 cytokines if the T cells are derived from atopic donors. This phenomenon is not due to diminished IL-12 production by DCs of atopic donors. PMID- 10808182 TI - Rapid oral challenge-desensitization for patients with aspirin-related urticaria angioedema. AB - BACKGROUND: Acetylsalicylic acid (ASA), commonly known as aspirin, is indicated in the treatment of coronary artery disease (CAD). Many patients are denied treatment with ASA because of a history of ASA or nonsteroidal anti-inflammatory drug (NSAID)-induced urticaria or angioedema. OBJECTIVE: We sought to develop a safe and practical protocol to allow the administration of ASA to patients with a history of ASA- or NSAID-induced urticaria-angioedema. METHODS: Eleven subjects with a history of ASA- or NSAID-induced urticaria-angioedema were challenged desensitized by oral protocols based on rapidly escalating doses of ASA. Most had CAD, one had a history of pulmonary embolism, and one had refractory chronic sinusitis and asthma. Starting doses ranged from 0.1 to 10 mg and were administered at intervals of 10 to 30 minutes. Dosing was individualized for each patient but followed this general sequence (in milligrams): 0.1, 0.3, 1, 3, 10, 20, 40, 81, 162, 325. RESULTS: Nine patients tolerated the procedure without adverse effects and continued taking ASA for periods ranging from 1 to 24 months, without development of urticaria or angioedema. A patient who had a history of chronic idiopathic urticaria in addition to aspirin-induced urticaria had chest tightness during the protocol. Another patient who had continuing urticaria and angioedema associated with antithyroid antibodies developed angioedema several hours after completing the protocol. CONCLUSION: In patients with historical ASA- or NSAID-induced urticaria-angioedema reactions but who did not have urticaria and angioedema independent of ASA/NSAID, rapid oral challenge-desensitization to ASA was performed safely and permitted patients with CAD and other diseases to receive treatment with ASA. PMID- 10808183 TI - Evidence for increased expression of eotaxin and monocyte chemotactic protein-4 in atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease associated with tissue eosinophilia and the activation of T lymphocytes. The novel eosinophil chemoattractants, eotaxin and monocyte chemotactic protein (MCP) 4, are up-regulated at sites of allergic inflammation, yet their contribution to the pathophysiologic mechanisms of AD remains to be determined. OBJECTIVE: We sought to investigate the expression of eotaxin and MCP-4 in acute and chronic lesions from patients with AD and to determine their relationship to the numbers of resident inflammatory cells. METHODS: With use of in situ hybridization, the expression of eotaxin and MCP-4 messenger RNA (mRNA) in skin biopsy specimens from patients with acute and chronic AD skin lesions was compared with that of uninvolved skin from these patients and skin from healthy volunteers. RESULTS: There was a constitutive expression of eotaxin and MCP-4 mRNA in skin biopsy specimens from healthy subjects. Positive signal for chemokine mRNA was observed both within the epidermis and inflammatory cells (macrophages, eosinophils, and T cells) of the subepidermis in AD skin lesions. Within the subepithelium acute and chronic skin lesions exhibited a significant increase in the numbers of eotaxin and MCP-4 mRNA-positive cells compared with uninvolved skin (P <.01), whereas the numbers of eotaxin and MCP-4 mRNA-positive cells were significantly higher in chronic AD compared with acute AD skin lesions (P <.005, P <.001, respectively). Correlations were observed between the expression of eotaxin and MCP-4 mRNA and the presence of eosinophils and macrophages, respectively, in AD lesions (r(2) = 0.84, r(2) = 0.94). CONCLUSION: There is an increased expression of eotaxin and MCP-4 in acute and chronic lesions, suggesting that these chemotactic factors play a major role in the pathophysiologic mechanisms of AD. PMID- 10808184 TI - Clinical and immunologic variables in skin of patients with atopic eczema and either positive or negative atopy patch test reactions. AB - BACKGROUND: Epicutaneous application of aeroallergens induces a positive atopy patch test (APT) response in about 50% of patients with atopic eczema (AE) and sensitization for these allergens. OBJECTIVE: To elucidate the mechanisms determining the outcome of the APT, the following questions were addressed. Are there differences in clinical features between patients with AE who have positive versus negative APT responses? Is a macroscopically negative APT response also histologically negative, and if so, are there differences in clinically noninvolved skin between the two groups regarding (1) the sensitivity toward an irritant, (2) the composition of cellular infiltrate, (3) the presence of aeroallergen-specific T cells, and (4) the number of IgE(+) cells? METHODS: Punch biopsy specimens from both house dust mite patch tested and the clinically noninvolved skin of patients with AE who have positive APT responses (n = 10) and negative APT responses (n = 10) and those from the normal skin of atopic individuals without AE (n = 10) and nonatopic volunteers (n = 10) were analyzed by using immunohistochemistry with mAbs against eosinophil cationic protein, IgE, the high-affinity receptor for IgE, and CD3 and CD25 mAbs. Furthermore, T-cell lines were propagated from noninvolved skin of all patient and control groups. The T-cell lines were tested for house dust mite specificity. RESULTS: Negative APT sites were immunohistochemically similar to clinically noninvolved AE skin. There were no significant differences between patients with AE who had positive and negative APT results regarding either clinical features, the composition of cellular infiltrate, or the presence of allergen-specific T cells in clinically noninvolved skin. However, differences were observed regarding the presence of IgE on epidermal CD1a(+) cells. CONCLUSION: Our results indicate that a positive APT reaction requires the presence of epidermal IgE(+) CD1a(+) cells in clinically noninvolved skin, but that also other, as yet unknown, discriminatory factors are involved. PMID- 10808185 TI - Human T-cell epitopes of the latex allergen Hev b 5 in health care workers. AB - BACKGROUND: Latex allergy affects health care workers as a high-risk cohort. Hev b 5 is a major latex allergen reacting with serum IgE from 92% of latex-allergic health care workers. Because CD4(+) T-cell recognition is central to the specific immune response to allergens, identification of dominant T-cell epitopes is important for the development of specific immunotherapy for latex allergy. OBJECTIVE: Our purpose was to map T-cell epitopes of Hev b 5 in health care workers. METHODS: Six latex-allergic health care workers (grade 3 to 4 enzyme allergosorbent test score) were studied. Peripheral blood latex specific 3-week T cell lines were generated and screened for proliferative response to overlapping 20-mer peptides of Hev b 5. Supernatants collected at 48 hours were analyzed by ELISA for IL-5 and IFN-gamma. RESULTS: Dot immunoblotting with use of recombinant Hev b 5/maltose-binding protein indicated serum-specific IgE in 5 of 6 patients. T-cell reactivity to one or more Hev b 5 peptides was identified in these 5 donors, but not in the sixth. Hev b 5 (46-65) induced T-cell proliferation in all 5 donors. Hev b 5 (109-128) stimulated T cells from 3 of these patients. Proliferative responses were accompanied by substantial IL-5 secretion and minimal IFN-gamma, indicating a T(H)2-predominant cytokine profile. CONCLUSIONS: Five of 6 latex-allergic patients demonstrated T-cell responsiveness to Hev b 5 consistent with a major T-cell reactive latex allergen. Two T-cell immunodominant regions of Hev b 5 were identified, and reactivity to these sites was associated with strong IL-5 but minimal IFN-gamma production. PMID- 10808186 TI - Outdated EpiPen and EpiPen Jr autoinjectors: past their prime? AB - BACKGROUND: EpiPen and EpiPen Jr autoinjectors are often recommended for prehospital treatment of anaphylaxis. When these units become outdated, there may be a delay in replacing them. OBJECTIVES: Our purpose was to evaluate unused, outdated EpiPen and EpiPen Jr autoinjectors, obtained from patients at risk for anaphylaxis, for epinephrine bioavailability and epinephrine content. METHODS: We conducted a prospective, randomized, cross-over study of epinephrine bioavailability after injection from outdated autoinjectors in rabbits; controls included EpiPen and EpiPen Jr autoinjectors that had not expired ("in-date" autoinjectors) and intramuscular injection of 0.9% saline solution. In addition, the epinephrine content of the outdated EpiPen and EpiPen Jr autoinjectors was measured by a spectrophotometric method and an HPLC-UV method. RESULTS: Twenty eight EpiPen and 6EpiPen Jr autoinjectors were studied 1 to 90 months after the stated expiration date. Most were not discolored and did not contain precipitates. Epinephrine bioavailability from the outdated EpiPen autoinjectors was significantly reduced (P <.05) compared with epinephrine bioavailability from the in-date autoinjectors. The inverse correlation between the decreased epinephrine content of the outdated autoinjectors, assessed with an HPLC-UV method, and the number of months past the expiration date was 0.63. CONCLUSIONS: For prehospital treatment of anaphylaxis, we recommend the use of EpiPen and EpiPen Jr autoinjectors that are not outdated. If, however, the only autoinjector available is an outdated one, it could be used as long as no discoloration or precipitates are apparent because the potential benefit of using it is greater than the potential risk of a suboptimal epinephrine dose or of no epinephrine treatment at all. PMID- 10808187 TI - Allergenicity of mare's milk in children with cow's milk allergy. AB - BACKGROUND: Cow's milk allergy is a common disease of infancy and early childhood. If the baby is not breast-fed, a substitute for cow's milk formula is necessary. OBJECTIVE: The aim of this study was to investigate, in vitro and in vivo, the allergenicity of mare's milk in a population of selected children with severe IgE-mediated cow's milk allergy. METHODS: Twenty-five children (17 male and 8 female) aged 19 to 72 months (median age 34 months) with IgE-mediated cow's milk allergy were selected for this study. All the children underwent skin prick tests with cow's milk and mare's milk and double-blind placebo-controlled oral food challenge (DBPCOFC) with fresh cow's milk, fresh mare's milk, and, as placebo, a soy formula (Isomil, Abbott, Campoverde, Italy). We performed immunoblotting of cow's and mare's milk developed with IgE from allergic children. RESULTS: All the children showed strong positive skin test responses to cow's milk (4+); 2 children had positive skin test responses to mare's milk (2+). All children had positive DBPCOFCs to cow's milk; one child had a positive DBPCOFC to mare's milk. No children reacted to the placebo (Isomil). In the cow's milk, some proteins are able to strongly react with human IgE; when the sera are tested with mare's milk, the bands corresponding to the same proteins are recognized by a lower percentage of sera. CONCLUSION: These data suggest that mare's milk can be regarded as a good substitute of cow's milk in most children with severe IgE-mediated cow's milk allergy. It would be prudent, however, to confirm its tolerability by a supervised titrated oral challenge test. PMID- 10808188 TI - Treatment with IFN-alpha in corticosteroid-unresponsive asthma. PMID- 10808191 TI - Editorial. AB - As we begin the new millennium, it is a great privilege for me to assume the responsibility of Editor-in-Chief of Endocrine-Related Cancer. As members of the Society for Endocrinology will know, Endocrine-Related Cancer< represents the further development and expansion of Reviews on Endocrine-Related Cancer which was acquired by the Society in 1993. Under the brilliant stewardship of its founding editor, Vivian James, Endocrine-Related Cancer has grown to be an outstanding journal publishing significant reviews in the area of intersection between endocrinology and cancer as well as proceedings of important scientific meetings and symposia around the world. PMID- 10808192 TI - New pituitary oncogenes. AB - Pituitary tumors are common monoclonal neoplasms which cause considerable morbidity and mortality. Several molecular events underlying pituitary tumorigenesis have been elucidated in recent years, but no tumor marker has clearly emerged which assists clinical and therapeutic decisions. Activating mutations and loss of inactivating mutations, together with hypothalamic hormones, circulating hormones, growth factors and cytokines cooperatively ensure the inexorable expansion of the initial mutated pituitary cell clone. This review describes new developments in our understanding of the molecular mechanisms involved in the pathogenesis of pituitary tumors. The availability of molecular probes will allow the early prediction of tumor behavior, identify targets for designing subcellular pituitary tumor therapy and provide novel approaches to pituitary tumor management. PMID- 10808193 TI - Similarities and distinctions in the mode of action of different classes of antioestrogens. PMID- 10808194 TI - The epidemiology of endocrine tumours. PMID- 10808195 TI - The role of prostate specific antigen measurement in the detection and management of prostate cancer. AB - The introduction of prostate specific antigen (PSA) testing has revolutionised the early detection, management and follow-up of patients with prostate cancer and it is considered to be one of the best biomedical markers currently available in the field of oncology. Its use with annual digital rectal examination in prostate cancer screening programmes has led to a marked change in the distribution of stage at presentation towards earlier disease and led to a significant increase in the detection of potentially curable disease. In order to improve the specificity of PSA testing and thereby reduce the number of unnecessary prostatic biopsies, a number of refinements of PSA evaluation have been proposed. These include free to total PSA ratio, PSA density, PSA density, PSA density of the transition zone, PSA velocity and age-specific PSA reference ranges. The utility of these approaches is considered in this review. The role of PSA monitoring in the detection of recurrence following radical prostatectomy and radiotherapy is discussed, as well as its role in monitoring patients treated with endocrine therapy is discussed, as well as its role in monitoring patients treated with endocrine therapy in terms of correlating PSA response with outcome, in detecting disease progression and in guiding the use of subsequent therapies. Large continuing multicentre screening and outcome studies will provide important information enabling greater refinement of the use of this important diagnostic and monitoring tool in the future detection and management of prostate cancer. PMID- 10808196 TI - Hormone replacement therapy and breast cancer. AB - The concern that postmenopausal hormone replacement therapy (HRT) may cause cancer of the breast has lead to an enormous volume of research in epidemiology, endocrinology and tumour cell biology. The epidemiology has become extremely sophisticated because the anticipated effect is small and there are several confounding factors. The consensus today is that long-term HRT (>10 years) is associated with an increase in the risk of breast cancer which, on average, is equivalent to delaying menopause for the same period of time that the patient is on treatment. The risk is related to endogenous and exogenous oestrogen levels. Studies that have investigated individual susceptibility are reviewed, as are environmental factors such as the interaction of HRT with alcohol intake. The clinical implication of these data is that the dosage of HRT should be the smallest that is efficacious. Subcutaneous implants of oestrogen typically cause very high oestrogen levels and, in the opinion of this reviewer, should be restricted to women unable to take or absorb oestrogen by mouth or percutaneously. Finally, the issue of HRT for women with a history of breast cancer is considered. The potential is discussed for treatment of women with severe symptoms of oestrogen deficiency with a low dose of oestrogen, together with a selective oestrogen receptor modulator to protect the breast. PMID- 10808197 TI - Factors released by rat type 1 astrocytes exert different effects on the proliferation of human neuroblastoma cells (SH-SY5Y) in vitro. AB - Brain metastases derived from abdominal neuroblastoma are an uncommon complication of this tumour; however, an increase in their occurrence has recently been reported. In the present study, we have investigated the influence of factors derived from central nervous system glial cells on the proliferation of human neuroblastoma cells (SH-SY5Y) in vitro. Co-culture experiments show that a 24-h exposure to factors released by type 1 astrocytes (A1) may induce a significant decrease in [(3)H]thymidine ([(3)H]TdR) incorporation by SH-SY5Y cells. This effect was not duplicated by fresh A1-conditioned medium (A1-CM); A1 CM became active only when it was heated or frozen. In contrast to this short lived inhibitory effect, long-term treatment (3, 6 and 9 days) with A1-CM produced a significant and dose-dependent increase in SH-SY5Y cell number. Immunoneutralisation of A1-CM with an anti-transforming growth factor-beta antibody eliminated the inhibitory effect on [(3)H]TdR uptake in SH-SY5Y cells, but did not affect the increased number of viable cells observed after long-term treatments. In conclusion, these results showed that factor(s) released by A1 may affect the proliferation/survival of a human neuroblastoma cell line in vitro inducing: (a) a short transient negative effect on DNA synthesis and (b) an overall sustained trophic action. These results are suggestive of a possible role of glial cells in the establishment of brain metastases of neuroblastomas. PMID- 10808198 TI - Can science and politics really be divorced? PMID- 10808199 TI - Hematopoietic stem cell transplants after reduced intensity conditioning regimen (RI-HSCT): report of a workshop of the European group for Blood and Marrow Transplantation (EBMT). AB - Allogeneic minitransplants are being increasingly used for the treatment of hematologic malignancies. Eligibility criteria usually include age over 60 years and/or major clinical problems concomitant with the hematologic disorder. This report summarizes a workshop which was held in 1999 under the auspices of EBMT: the issues discussed included stem cell sources and mobilization techniques, stem cell engraftment, accessory cells, mixed chimerism, graft-versus-tumor effect and ongoing clinical trials. There are many questions which still need to be answered, but this area of transplantation seems very promising and will certainly develop in the coming years. PMID- 10808200 TI - A dose escalation study of total body irradiation followed by high-dose etoposide and allogeneic blood stem cell transplantation for the treatment of advanced hematologic malignancies. AB - Since approximately 30% of leukemia patients relapse after allogeneic BMT using total body irradiation (TBI)-based preparative regimens, treatment intensity may be suboptimal. The killing of leukemia cells is proportional to the radiation absorbed dose. We studied the feasibility and toxicity of escalating the doses of fractionated TBI above our previous prescription of 13.5 Gy. Sixteen evaluable patients with advanced hematologic malignancies were treated with twice daily TBI using a high-energy source (18-24 MV). The first patient cohort (n = 11) received a total dose of 14.4 Gy in nine fractions, and the second cohort (n = 5) received doses escalated to 15.3 Gy. All patients received high-dose etoposide (60 mg/kg) and allogeneic stem cell transplantation following the TBI. All patients had HLA identical sibling donors. The median times for neutrophil and platelet engraftment were 13.5 and 12 days, respectively, and did not differ between the two cohorts. All but one patient developed treatment-related grade 3 or 4 mucositis. There were three cases of grade 4 pulmonary toxicity and three cases of grade 4 hepatic toxicity among the 14.4 Gy cohort, and one case each of grade 4 pulmonary and hepatic toxicities among the 15.3 Gy cohort. In most cases comorbid conditions contributed to these toxicities. Two patients had significant GVHD of the GI tract. Six relapses occurred, five (45%) in the 14.4 Gy cohort and one (20%) in the 15.3 Gy cohort. The 100-day treatment-related mortality rates were 9% and 20% for the 14.4 Gy and 15.3 Gy cohorts, respectively, and the median survivals were 226 and 201 days, respectively. We conclude that TBI dose escalation above the previously used 13.5 Gy dose is feasible using a high-energy source and high-dose etoposide. Acute and chronic toxicities were primarily related to GVHD, infection and relapse rather than to TBI. PMID- 10808201 TI - Bone marrow transplantation from alternative donors for thalassemia: HLA phenotypically identical relative and HLA-nonidentical sibling or parent transplants. AB - Twenty-nine patients with thalassemia and a median age of 6 years (range 1.1-33 years) were given a BMT from an alternative donor. Six of the 29 donors were HLA phenotypically identical and two were mismatched relatives, 13 were mismatched siblings and eight were mismatched parents. Six patients received no antigen (relatives), 15 patients one antigen, five patients two antigen and three patients three antigen disparate grafts. Twenty-three patients were in class 2 or class 3, whereas six patients were in class 1. Thirteen patients were given BUCY, nine patients BUCY plus ALG, six patients BUCY plus TBI or TLI and one patient BUCY with prior cytoreductive-immunosuppressive treatment as conditioning. As GVHD prophylaxis four patients received MTX, 22 CsA + MTX + methylprednisolone (MP) and three patients CsA + MP. Thirteen of 29 patients (44.8%) had sustained engraftment. The probability of graft failure or rejection was 55%. There were no significant differences between antigen disparities and graft failure. The incidence of grade II-IV acute GVHD was 47.3% and chronic GVHD was 37.5%. The incidence of acute GVHD was higher in patients receiving one or two antigen disparate in the GVHD direction grafts (vs no antigen) (P EQ 0.04; odds ratio 10.8; 95% CI 1.5-115). The probability of overall and event-free survival was 65% and 21%, respectively, with median follow-up of 7.5 years (range 0.6-17 years) for surviving patients. The degree of HLA disparity between patient and donor did not have a significant effect on survival. The incidence of nonhematologic toxicity was low. Transplant-related mortality was 34%. GVHD (acute or chronic) was a major contributing cause of death (50%) followed by infections (30%). We conclude that at present, due to high graft failure and GVHD rates, BMT from alternative donors should be restricted to patients who have poor life expectancies because they cannot receive adequate conventional treatment or because of alloimmunization to minor blood antigens. PMID- 10808203 TI - Purging of G-CSF-mobilized peripheral autografts in acute leukemia with mafosfamide and amifostine to protect normal progenitor cells. AB - In the present study the in vitro growth of CFU-GM from PBPC of patients with AML (n = 11), purged with mafosfamide alone or a combination of mafosfamide and amifostine, was compared to historical controls of mafosfamide-purged bone marrow (AML CR1, n = 16). Two patients were transplanted with mafosfamide and mafosfamide/amifostine pretreated PBPC autografts. The in vitro experiments demonstrated a significantly higher resistance of peripheral blood derived CFU-GM to mafosfamide (median ID95 190 microg mafosfamide/ml) compared with bone marrow derived CFU-GM (median ID95130 microg/ml). Preincubation with amifostine significantly further increased the median ID95 to 245 microg/ml. The clinical results showed short recovery times for neutrophils >500/microl (9 and 13 days) and platelets >20 000/microl (12 and 21 days) and stable long-term engraftment with one relapse at day +118 and one patient in CR at day 760 after transplantation. The in vitro results show a significant advantage of PBPC over bone marrow-derived progenitors for purging with mafosfamide. Furthermore, a protective effect from mafosfamide of amifostine on normal progenitors could be demonstrated. The clinical results demonstrate the clinical feasibility of using mafosfamide-purged autologous PBPCT without impairing the short-term and long term repopulating capacities of the autografts. PMID- 10808202 TI - Autologous bone marrow transplantation for acute myeloid leukemia in remission or first relapse using monoclonal antibody-purged marrow: results of phase II studies with long-term follow-up. AB - One hundred and thirty-eight patients with AML underwent ABMT with monoclonal antibody plus complement-purged marrow between August 1984 and March 1997. One hundred and ten patients were in CR (CR1: 23; CR2/3: 87) and 28 were in first relapse (R1) at ABMT. Preparative regimens included busulfan (16 mg/kg) and CY (120 mg/kg) (n = 93), CY (120 mg/kg over 2 days) with TBI (1200 cGy) (n = 35), and busulfan (16 mg/kg) plus etoposide (60 mg/kg) (n = 10). CR1 patients treated with CY/TBI (n = 7) had 3- and 5-year disease-free survival (DFS) rates of 71% and 57%. CR1 patients treated with BU/CY (n = 12), had 3- and 5-year DFS rates of 45%. Three and 5-year DFS for CR2/3 patients treated with CY/TBI (n = 26) was 23%. Three- and 5-year DFS for patients in CR2/3 treated with BU/CY (n = 55) was 31 and 28%. Three- and 5-year DFS for patients in R1 treated with BU/CY (n = 26) was 37%. In multivariate analysis, increased age was associated with greater risk of death and relapse. For CR2/3 patients, the length of CR1 was a significant predictor of DFS. ABMT performed in CR or R1 results in excellent 5-year DFS and OS. The contribution of purging may require a randomized trial comparing purged vs unpurged stem cell infusions. PMID- 10808204 TI - Histological alterations in bone marrow in patients with late engraftment after autologous bone marrow transplantation. AB - Bone marrow histology after bone marrow transplantation has rarely been studied. Here, we reviewed the pre- and post-transplant bone marrow biopsies (BMB) of 40 acute myelogenous leukemia (AML) patients autografted in our center, 28 with normal and 12 with delayed peripheral recovery. The two groups were comparable in terms of previous therapy, disease phase and the number of infused cells, and received the same conditioning regimen. In the former group, reduced bone marrow cellularity and mild reticulin abnormalities were usual histological findings; in the latter, five patients had the same pattern, but the other seven had an almost undetectable hematopoietic parenchyma and severe reticulin derangement. One of these seven patients died of reactivated hepatitis B virus infection; the others eventually achieved peripheral recovery, with none of them experiencing a relapse. Autografted AML patients are excellent subjects for histological investigations. They account for the majority of delayed engraftments, the contribution of extramedullary components to the timing of engraftment is minimal, and leukemia relapse cannot be ruled out. These results suggest that BMB is a useful investigation in the work-up of late engraftment. A high degree of reticulin derangement with an almost undetectable hematopoietic parenchyma appear to be the morphological hallmarks of late engraftment. PMID- 10808205 TI - Mixed chimerism in the B cell lineage is a rapid and sensitive indicator of minimal residual disease in bone marrow transplant recipients with pre-B cell acute lymphoblastic leukemia. AB - One of the major problems after allogeneic bone marrow transplantation (BMT) is a high frequency of leukemia relapse. We have prospectively studied the presence of donor- and recipient-derived chimeric cells in bone marrow recipients with pre-B cell acute lymphoblastic leukemia (pre-B-ALL). The chimeric status of BMT recipients was compared to minimal residual disease (MRD) detection by analysis of immunoglobulin heavy chain (IgH) and T cell receptor (TcR) genes. Post transplant blood and bone marrow samples from 12 patients with pre-B-ALL were studied. Five patients showed mixed chimerism (MC) in the CD19-positive cell fraction. Four of them have relapsed to date. The remaining patient with MC in the B cell lineage was also MRD positive in the same samples. All seven patients with donor chimerism in the B cell fraction remain in clinical remission (P = 0.01). In samples from all five patients having MC in the B cell lineage, the patient-specific IgH or TcR rearrangement was also detected. In three of four patients who relapsed, MC in the B cell lineage was seen more than 2.5 months prior to morphologically verified relapse. The results of this comparison suggest that routinely performed MC analysis of the affected cell lineage may facilitate post-BMT monitoring and rapid therapeutic decisions in transplanted patients with pre-B-ALL. PMID- 10808206 TI - Fluconazole vs low-dose amphotericin B for the prevention of fungal infections in patients undergoing bone marrow transplantation: a study of the North American Marrow Transplant Group. AB - Systemic fungal infections are a major problem in bone marrow transplant recipients who have prolonged neutropenia or who receive high-dose corticosteroids. Prophylaxis with Fluconazole or low-dose amphotericin B reduces, but does not eliminate these infections. To determine which prophylactic agent is better, we performed a prospective randomized study. Patients undergoing allogeneic (related or unrelated) or autologous marrow or peripheral stem cell transplantation were randomized to receive Fluconazole (400 mg/day p. o. or i.v.) or amphotericin B (0.2 mg/kg/day i.v.) beginning 1 day prior to stem cell transplantation and continuing until recovery of neutrophils to >500/microl. Patients were removed from their study drug for drug-associated toxicity, invasive fungal infection or suspected fungal infection (defined as the presence of fever >38 degrees C without positive culture while on broad-spectrum anti bacterial antibiotics). Proven or suspected fungal infections were treated with high-dose amphotericin B (0.5-0.7 mg/kg/day). Patients were randomized at each institution and stratified for the type of transplant. The primary end-point of the study was prevention of documented fungal infection; secondary endpoints included fungal colonization, drug toxicity, duration of hospitalization, duration of fever, duration of neutropenia, duration and total dose of high-dose amphotericin B and overall survival to hospital discharge. From July 1992 to October 1994, a total of 355 patients entered into the trial with 159 patients randomized to amphotericin B and 196 to Fluconazole. Patient groups were comparable for diagnosis, age, sex, prior antibiotic or antifungal therapy, use of corticosteroids prior to transplantation and total duration of neutropenia. Amphotericin B was significantly more toxic than Fluconazole especially in related allogeneic transplantation where 19% of patients developed toxicity vs 0% of Fluconazole recipients (p < 0.05). Approximately 44% of all patients were removed from prophylaxis for presumed fungal infection. Proven fungal infections occurred in 4.1% and 7.5% of Fluconazole and amphotericin-treated patients, respectively. Proven fungal infections occurred in 9.1% and 14.3% of related allogeneic marrow recipients receiving Fluconazole or amphotericin B, respectively, and 2.1% and 5.6% of autologous marrow recipients receiving Fluconazole or amphotericin B, respectively (P > 0.05). In this prospective trial, low-dose amphotericin B prophylaxis was as effective as Fluconazole prophylaxis, but Fluconazole was significantly better tolerated. PMID- 10808207 TI - Association of cytomegalovirus interstitial pneumonitis with HLA-type following allogeneic bone marrow transplantation. AB - Certain human leukocyte antigens may increase the risk of cytomegalovirus interstitial pneumonitis, an important complication of bone marrow transplantation. The prevalence of this pneumonitis was compared between patients possessing either HLA-B51 or HLA-B52 and patients without either antigen. The role of tumor necrosis factor-alpha in cytomegalovirus interstitial pneumonitis was also studied. Among 72 patients undergoing allogeneic bone marrow transplantation at our institution during the past 5 years, HLA-B51 or -B52 was detected in 29. Among these 29 patients, 13 (45%) developed cytomegalovirus interstitial pneumonitis, a significantly higher rate (P < 0.001) than among patients without these HLA types (4/43, 9%). In the pre-conditioning and stable phases, tumor necrosis factor-alpha levels were higher in patients with HLA-B51 or HLA-B52 than in patients without (P < 0.05; t-test). Throughout the period from pre-conditioning to around day 40, except on day 0, tumor necrosis factor alpha levels were also significantly higher (P < 0.05 to P < 0.001) in patients developing cytomegalovirus infection than in those without it. These results suggest that HLA-B51 and HLA-B52 may be risk factors for cytomegalovirus interstitial pneumonitis after bone marrow transplantation, with an increase of tumor necrosis factor-alpha also being involved. PMID- 10808208 TI - Diagnostic aspects of invasive Aspergillus infections in allogeneic BMT recipients. AB - To investigate diagnostic aspects of invasive aspergillosis (IA) in allogeneic BMT recipients, the charts of 22 consecutive patients with IA transplanted in 1989-1995 were reviewed. IA was diagnosed 69-466 days (median 131 days) post BMT. In 16 patients (73%), a definite or probable diagnosis of IA was made during life. Respiratory symptoms were the presenting feature in half of the patients followed by neurological symptoms (27%). Chest X-ray revealed single or multiple nodular lesions in 10 patients; cavitation was observed in five patients. Tissue biopsy was the most common method of diagnosis (nine patients: lungs 6, liver 1, subcutaneous tissue 1, brain 1). Five IA cases were detected by nine guided fine needle lung biopsies in eight patients and without complications. Bronchoalveolar lavage was performed in 14 patients with findings suggestive of invasive pulmonary aspergillosis in eight cases. Lungs were the most common organ affected (90%) followed by central nervous system (41%). The diagnosis of IA is still difficult, and a large number of patients have advanced infection at diagnosis. Methods for early diagnosis are needed. Patients with a clinical suspicion of IA should be treated vigorously with antifungal agents during the diagnostic work up. PMID- 10808209 TI - An outbreak of multiply resistant Serratia marcescens: the importance of persistent carriage. AB - An outbreak of multi-resistant Serratia marcescens involving 24 patients occurred in a bone marrow transplant and oncology unit, from September 1998 to June 1999, of whom 14 developed serious infection. This is the first such outbreak described in a BMT unit. All isolates demonstrated the same antimicrobial susceptibility pattern and were the same unusual serotype O21:K14. The antimicrobial susceptibility profile showed reduced susceptibility to ciprofloxacin, gentamicin and piperacillin-tazobactam. As the latter two antimicrobials are part of our empiric therapy for febrile neutropenia, they were substituted with meropenem and amikacin during the outbreak. Investigation revealed breaches in infection control practices. Subsequently, the outbreak was contained following implementation of strict infection control measures. A prominent feature of the outbreak was prolonged carriage in some patients. These patients may have acted as reservoirs for cross-infection. This report also indicates that patients who become colonised with Serratia marcescens may subsequently develop invasive infection during neutropenic periods. PMID- 10808210 TI - Amphotericin B colloidal dispersion (Amphocil) vs fluconazole for the prevention of fungal infections in neutropenic patients: data of a prematurely stopped clinical trial. AB - We conducted an open label, randomised clinical trial to compare amphotericin B colloidal dispersion (ABCD, Amphocil) 2 mg/kg/day intravenously with fluconazole 200 mg/day orally, for the prevention of fungal disease in neutropenic patients with haematological malignancies. In the event of unresolved fever after 4 days of empirical antibacterial therapy, patients in both treatment groups were to receive ABCD, 4 mg/kg/day. However, the study had to be stopped in an early phase, due to severe side-effects of ABCD. A total of 24 patients were enrolled, 12 patients were randomly assigned to receive prophylactic ABCD, which was administered for a mean of 13.9 days. Fluconazole prophylaxis was given to 12 patients for a mean of 21.2 days. Therapeutic ABCD, 4 mg/kg, was initiated in four patients because of suspected fungal infection, all of whom had initially received fluconazole. A high rate of infusion-related toxicity of ABCD was observed. Chills occurred in 15/16 ABCD recipients (94%), accompanied by a temperature rise of >/=2 degrees C in 4/16 patients and of >/=1 degrees C but <2 degrees C in 10/16 patients. Other ABCD-related adverse events were hypotension (4/16), nausea with vomiting (5/16), tachycardia (7/16), headache (3/16) and dyspnoea (3/16). For premedication patients received: antihistamines (12/16), hydrocortisone (9/16) and/or morphine (6/16). ABCD was discontinued in 8/16 patients (50%) due to side-effects, which ultimately dictated early termination of the study. We conclude that ABCD is not suitable for antifungal prophylaxis in neutropenic patients due to severe infusion-related side-effects. Subject numbers were too low for conclusions on variables of antifungal efficacy. PMID- 10808211 TI - Cardiac and pulmonary toxicity in patients undergoing high-dose chemotherapy for lymphoma and breast cancer: prognostic factors. AB - We sought to define risk factors predisposing breast cancer and lymphoma patients to cardiac and pulmonary toxicity when undergoing high-dose chemotherapy (HDC) and autologous stem cell rescue (ASCR). Additionally, we evaluated in depth the predictive value of the ejection fraction measured prior to HDC in determining cardiac toxicity. In this retrospective analysis, 24 variables were examined in 138 patients undergoing HDC and ASCR from 1990 until 1995. Logistic regression models were used to model the probability of experiencing cardiac and pulmonary toxicity as a function of the 24 prognostic covariates. Cardiac toxicity occurred in 12% of patients and pulmonary toxicity in 24% of patients. Bivariate analyses showed that patients with lymphoma (as opposed to breast cancer) and those with a higher cardiac risk factor score were more likely to experience cardiac toxicity. Multivariate logistic regression models predicted lymphoma and older age to be risk factors for cardiac toxicity. History of an abnormal ejection fraction and higher doses of anthracyclines prior to HDC may also contribute to cardiac toxicity. Pulmonary toxicity occurred more commonly in lymphoma than breast cancer patients, likely due to the busulfan used in the HDC regimen. No other risk factors for pulmonary toxicity were identified. We conclude that older patients with lymphoma should be carefully evaluated prior to being accepted for HDC programs. Older patients with breast cancer may tolerate this procedure well. There is a trend towards cardiac toxicity in patients with a past history of low ejection fraction, although seemingly poor cardiac risk patients may fare well with HDC if carefully selected with the aid of a thorough cardiac evaluation. PMID- 10808212 TI - Treatment of solid tumors following allogeneic bone marrow transplantation. AB - Second solid tumors are well known late complications after bone marrow transplantation. Treatment strategies are ill defined. We retrospectively evaluated treatment and outcome in a single institution. From August 1974 to July 1996, six solid tumors were observed in five of 387 patients 2 to 13 years after BMT, corresponding to a probability of developing a second solid tumor of 9% (1 17%, 95 CI) at 15 years: these comprised endometrial carcinoma, carcinoma of the thyroid gland, cervical carcinoma, sarcoma of the small intestine, osteosarcoma of the tibia and ovarian carcinoma. All five patients were treated as intensively as they would be without a history of BMT. At last follow-up four of the five patients were alive and without signs of tumor. We postulate that second solid tumors after BMT should be treated as de novo tumors. Early detection based on consequent clinical follow-up of the transplant patients might explain the relatively good outcome. PMID- 10808213 TI - Development of a district Cord Blood Bank: a model for cord blood banking in the National Health Service. AB - The Bristol Cord Blood Bank was established as a pilot project within existing health services to establish cost-effective recruitment, collection and processing suitable for use in the NHS should cord blood become a routine source of haemopoietic stem cells for transplantation in the UK. An important aim of the project was to evaluate the feasibility of establishing a midwifery-based collection network, thus utilising expertise already in place. Collection was performed on the delivery suite immediately after the placenta was delivered. The clinical experience of the midwife collector/counsellors allowed rapid pre collection assessment of the condition of the cord and placenta. This prevented collection attempts from diseased or otherwise damaged placentas, leading to conservation of resources by preventing collection of most small volume donations. The bank was established within the National Blood Service, Bristol Centre to achieve Good Manufacturing Practice standards and ensure that processing was subject to the same stringency required for other sources of haemopoietic stem cells. Cord blood is an expensive resource. By utilising existing expertise in district Obstetric and National Blood Services, the Bristol Cord Blood Bank may serve as a model for health economic evaluation of cord blood banking of volunteer donations within the NHS. PMID- 10808214 TI - Cross-sensitivity reaction between tacrolimus and macrolide antibiotics. AB - A patient with an allergy to a macrolide antibiotic was given tacrolimus and developed a sudden cutaneous reaction. PMID- 10808216 TI - Reimmunization after blood or marrow stem cell transplantation: reply to S Singhal and J Mehta. PMID- 10808215 TI - Donor bone marrow from a sibling with inborn error of metabolism for treatment of acute leukaemia - clinical and biochemical consequences in the non-affected recipient. AB - Bone marrow transplantation (BMT) is increasingly used in an attempt to correct inborn errors of metabolism (IEM). However, little is known about effects of BMT from patients with IEM donating for non-affected recipients. We present data from a 8.5-year-old girl who underwent BMT in second remission for relapsed acute lymphoblastic leukaemia (ALL) at the age of 7 years from her HLA-identical brother who was severely affected by Hunter syndrome (Mucopolysaccharidosis type II, iduronate-2-sulphatase (IDS) deficiency). After BMT not only leukocyte but also plasma activity of IDS was absent. Mixing experiments and immunoadsorption suggest antibody-mediated enzyme inhibition. However, her urinary glycosaminoglycan excretion has not increased post BMT and clinical signs of mucopolysaccharidosis are absent 20 months after BMT. We conclude that patients with white cell enzyme deficiencies and other IEMs do not have to be excluded from bone marrow donation. Antibody production by the graft may occur and be reflected by a marked reduction in plasma enzyme levels but not tissue activity. Similar antibody responses resulting in enzyme inactivation might also affect other enzyme replacement strategies for individuals with IEM. PMID- 10808217 TI - Interrelationship between cross-sectional and sagittal imaging in computer assisted dental implant tomography. AB - OBJECTIVE: To determine what changes may occur in computer-assisted dental tomography to cross-sectional and sagittal slices when the positions of one or both slices are adjusted. METHODS: A human skull with a ball bearing attached to the mandibular first molar was positioned off-centre in a CommCAT (Imaging Sciences International, Hatfield, PA, USA) tomographic machine. An occlusal projection of the mandibular dental arch was scanned into the computer. Correction measurements obtained from scout cross-sectional and sagittal slices were used to 'customize' cross-sectional and sagittal slices simultaneously and also individually. RESULTS: The resulting changes in position and blur of the ball bearing were analysed. When simultaneously and correctly customized, the resultant slices were at the correct locations with a sharp image of the marker. When the sagittal slice only was adjusted, the blur of the marker changed, but its position remained unchanged; at the same time, the blur of the marker in the cross-sectional slice remained unchanged, but its position changed. When the cross-sectional slice only was adjusted, the blur changed, but its position remained unchanged; at the same time, the blur of the marker in the sagittal slice remained unchanged, but its position changed. CONCLUSIONS: It is important for dentists evaluating tomograms for implant placement to be aware that with the CommCAT tomographic machine an interrelationship exists between cross-sectional and sagittal slices and that adjustment in one type of slice produces changes in either the blur or position of the marker in the adjusted and unadjusted slices. PMID- 10808218 TI - Subjective image quality of solid-state and photostimulable phosphor systems for digital intra-oral radiography. AB - OBJECTIVES: To compare subjectively the image quality of intra-oral radiographs from six digital systems. METHODS: Two generations of two different solid-state detectors; Visualix-1 and -2 (Gendex Dental Systems, Milan, Italy), Computed Dental Radiography (CDR) and CDR Active Pixel Sensor (APS) (Schick Technologies Inc., Long Island City, NY, USA), and two photostimulable phosphor (PSP) systems; Digora (Soredex, Orion Corporation, Helsinki, Finland) and DenOptix (Gendex Dental Systems, Milan, Italy) were compared. Tooth-containing specimens from different areas of dried mandibles were radiographed at exposures between 91-9400 microGy. Images were transferred to a personal computer, displayed in random order and evaluated in their original form and after applying a histogram equalisation algorithm. Eight observers graded subjective image quality using a 5 point scale. RESULTS: Both CDR systems scored highest for image quality but within the narrowest exposure range. The Visualix images received the lowest scores. The PSP systems produced acceptable image quality at both lower and higher exposures than the solid-state systems. Enhanced images were generally considered to be inferior to the original images, except for those produced by the four solid-state systems at very low exposures. CONCLUSIONS: (i) the PSP systems provided a clinically acceptable image quality over a wide exposure range; (ii) the CDR systems had the best image quality but over the narrowest exposure ranges; (iii) the Visualix systems had the lowest image quality; and (iv) histogram equalisation did not generally improve image quality. PMID- 10808219 TI - An objective comparison of four digital intra-oral radiographic systems: sensitometric properties and resolution. AB - OBJECTIVES: To compare the sensitometric properties and resolution of four digital intra-oral radiographic systems. METHODS: Three CCD-based systems were evaluated, RVG-4 (Trophy Radiologic, Paris, France) CDR (Schick Technologies Inc, Long Island, NY, USA) and Dixel (Morita, Kyoto, Japan) and a photostimulable phosphor system (Digora, Soredex-Orion, Helsinki, Finland). The sensitometric properties were determined by measuring the gray levels at increasing exposures without making any gray-scale adjustments. MTF was measured using images of a 10 microns width slit inclined to the long and short axes of the detectors. RESULTS: The CCD-based systems showed a linear relationship between exposure and gray level although there were some differences in sensitivity. In RVG-4, the gray level saturated at a lower exposure. Digora showed a logarithmic relationship at uniform exposure. However, there was a linear relationship when it was exposed below the calibration dose. The MTF was highest with RVG-4 and the lowest with the Digora. CONCLUSIONS: Each system has its own specific properties; RVG-4 has the highest resolution with the narrowest latitude, whereas Digora has the wider latitude but the lowest resolution. PMID- 10808220 TI - A clinical evaluation of some factors affecting image quality in panoramic radiography. AB - AIM: To evaluate the suitability of panoramic radiographs obtained with two different machines for clinical use. MATERIAL AND METHODS: Three observers assessed 111 pairs of panoramic radiographs taken with the Scanora (Soredex, Orion Corp., Helsinki, Finland) programme 001, and the Orthophos Plus (Sirona, Bensheim, Germany), programme P1 respectively, for visualisation of seven anatomical structures on each side, on a 5-point scale where 1 = very good. The median values were computed and a descriptive analysis performed. Interrater and intrarater agreement were expressed as weighted kappa. RESULTS: The median values of both Scanora and Orthophos radiographs were the same, scoring 2. The intra- and inter-observer agreement was generally poor and similar for both machines. CONCLUSIONS: The two panoramic machines were both given good ratings for evaluation of anatomical structures. The differences were very small and suggest that both machines are suitable for clinical use. PMID- 10808221 TI - Dental optical coherence tomography: a comparison of two in vitro systems. AB - OBJECTIVE: To compare the imaging results obtained with two different in vitro prototype dental optical coherence tomography (OCT) systems. METHODS: Two prototypes were evaluated: an 850 nm wavelength, 700 microW OCT system with a relatively low numerical aperture (0.03) and a 1310 nm wavelength, 140 microW system with a higher numerical aperture (0.20). RESULTS: Using the 850 nm system a characteristic scattering signal was observed that correlated with the depth of a periodontal probe. There was, however, insufficient light penetration to create images with adequate resolution. Improved image quality was achieved with the 1310 nm OCT system; these images had sufficient resolution to allow identification of anatomical structures important for the diagnostic assessment of oral structures. CONCLUSIONS: These results illustrate the improvement in imaging dental structures that can be obtained with a prototype 1310 nm OCT system. The feasibility of OCT as a dental imaging technique is verified. PMID- 10808223 TI - Digitisation and display of intra-oral films. AB - AIM: To determine the optimal parameters for the digitisation and display of intra-oral radiographs. MATERIALS AND METHODS: Six intra-oral radiographs (EP size 2 dental film, Eastman Kodak Co., Rochester, NY, USA) were digitised using an Arcus II image scanner (Agfa-Gevaert N. V., Mortsel, Belgium). Each film was digitised at 8 bits, with and without a film mask, using three scanner resolutions (200, 400 and 600 p.p.i.), resulting in six versions of each radiograph. Seven experienced observers were asked to evaluate the quality of the resulting images as they were displayed on a Nokia 445Xi monitor (Nokia Display Products, Helsinki, Finland). Monitor settings were varied by changing the colour palette and screen resolution. RESULTS: Digital images of masked films had a statistically significant (t-test with alpha 2 tail < or = 0.05) higher quality than those scanned without a mask. The best image quality was obtained at a resolution of 400 p.p.i. Display options were optimum with 256 colours and a screen size of 1152 x 864 pixels. CONCLUSIONS: Better results were obtained when intra-oral radiographs were digitised with a mask at a resolution of 200 or 400 p.p.i. and displayed using 256 colours and the largest screen size possible. Further studies are needed to extend these results to other makes and models of scanner and monitor. PMID- 10808222 TI - Sonographic analysis of rat submandibular glands in experimentally-induced sialadenitis. AB - OBJECTIVES: To correlate the sonographic features of experimentally-induced obstructive sialadenitis in rat submandibular glands with the histopathological changes. MATERIALS AND METHODS: Changes in sonograms of ligated and non-ligated rat submandibular glands were compared with the wet weight and histopathology. Sonograms were obtained at 1, 2, 3, 4, 5, 7 and 21 days after duct ligation at 13 and 14 MHz for B-mode and for power Doppler mode respectively. Changes in gland size, echo intensity, distribution of internal echoes and vascularity were evaluated both qualitatively and quantitatively. RESULTS: Changes in size of the ligated glands were very similar to changes in wet weight. The ligated glands showed lower echo intensity than the non-ligated glands. The ligated glands showed higher echo intensity in the central portion at 4-7 days after duct ligation which resulted in heterogeneity of the glands. Vascularity of the ligated glands increased in the acute phase, and then decreased to the level of the non-ligated glands. CONCLUSIONS: The three phases in the changes in ligated glands identifiable histopathologically can be differentiated by B-mode sonography. Power Doppler can differentiate the acute phase. Sonography therefore appears useful for the diagnosis of the degree of gland damage. PMID- 10808224 TI - The adherence to guidelines for facial radiography in an Accident & Emergency Department. AB - OBJECTIVES: To evaluate the degree of adherence to guidelines for facial radiography. METHODS: A retrospective review of 1538 facial radiographs from a consecutive series of 414 patients attending the Accident & Emergency Department of the North Staffordshire Hospital with suspected facial injuries over a 6 month period was undertaken. Information on age, sex and aetiology of injury was recorded along with the type of view and total number of films taken. The results were compared with guidelines from the Department of Oral & Maxillofacial Surgery. RESULTS: Almost one-half of all injuries were due to assault. There was general adherence to the guidelines in terms of the number of films taken for mid face and mandibular fractures. CONCLUSIONS: There was a good adherence to the guidelines in terms of the number of films taken for midface and mandibular fractures. However, a small number of patients were still being exposed to a high radiation dose where combinations of skull and facial views were taken. Guidelines need to be subjected to regular audit. PMID- 10808225 TI - Optimisation of patient doses in programmable dental panoramic radiography. AB - OBJECTIVES: To estimate the radiation-related risk associated with twelve imaging programs available on the Orthophos (Siemens, Erlangen, Germany) dental panoramic radiography unit. METHODS: Organ absorbed doses for each program were measured using a Rando anthropomorphic phantom loaded with thermoluminescent dosemeters. Effective dose (E) was calculated in two ways; first, using the method recommended by the International Commission on Radiological Protection, which excludes the salivary glands (designated Eexc), and second, with its inclusion (designated Einc). Organ and effective doses were both used to compare the various imaging programs. RESULTS: In 11 of the 12 programs studied the salivary glands received the highest individual organ dose, and Einc was found to be up to double Eexc. When the image was restricted to the dentition (program 2) organ doses were lower than for the complete jaws (program 1) by up to 85%, and Eexc and Einc reduced by about one half. When programs 2 and 6 (to image the temporomandibular joints) are used in place of program 1, the former combination provides more image information at an equivalent risk. CONCLUSIONS: The value of E in panoramic radiography depends on the inclusion of the salivary glands in the calculation and the magnitude of the dose. PMID- 10808226 TI - Three-dimensional morphology of the masseter muscle in patients with mandibular prognathism. AB - OBJECTIVE: To compare the morphology of the masseter muscle in patients with mandibular prognathism with that of normal subjects. METHODS: Three-dimensional X ray computed tomography (CT) was performed on 69 patients with mandibular prognathism and compared with 91 normal subjects. The angle of the muscle direction in relation to the Frankfurt horizontal plane and the area and the ratio of length of the short to long axes (s/l ratio) on the section perpendicular to the muscle direction were measured. RESULTS: The mean angle, area and s/l ratio in patients with mandibular prognathism was 76.6 degrees (s.d. 4.4 degrees), 318.3 mm2 (s.d. 77.2 mm2) and 0.312 (s.d. 0.049), respectively. Those of the normal subjects were 65.1 degrees (s.d. 4.4 degrees), 368.3 mm2 (s.d. 97.2 mm2) and 0.393 (s.d. 0.054), respectively. The angle was significantly larger, and the area and s/l ratio were significantly smaller than those of normal subjects (P < 0.001). CONCLUSION: The morphology of the masseter muscle in mandibular prognathism is significantly different from that of normal subjects. Our results may be helpful in evaluating the results of orthognathic surgery. PMID- 10808227 TI - Digital analysis of trabecular pattern in jaws of patients with sickle cell anemia. AB - OBJECTIVES: To characterize how the morphological features of the cancellous bone of the maxilla and mandible differ in patients with sickle cell anemia compared with normal subjects. METHODS: Periapical radiographs of 18 young African American patients with sickle cell anemia and 18 control African Americans were digitized at 600 d.p.i. A customized computer program measured 24 morphological features of the trabecular and marrow architecture in the anterior maxilla and mandible. The mean values for each of the features were determined for the sickle cell and control groups and compared by anatomical site. RESULTS: Patients with sickle cell anemia have a significant increase in area of the marrow (53% compared with 47% in controls) and a less complex trabecular structure as evidenced by fewer skeletal branch points per square cm (451 compared with 584 in controls in the anterior maxilla and, 553 compared with 678 in controls in the anterior mandible). All values are significantly different (P < 0.001). CONCLUSIONS: The data support the hypothesis that an automated image interpretation program can distinguish patients with sickle cell anemia from normal individuals. PMID- 10808229 TI - Tumoral calcinosis of the temporomandibular joint region. AB - A rare case of tumoral calcinosis, discovered in the medial pterygoid muscle and around the temporomandibular joint on a routine panoramic radiograph is presented. CT was found to be ideal for the determination of the exact location of the calcifications. The differential diagnosis of tumoral calcinosis is discussed. PMID- 10808228 TI - The value of magnetic resonance imaging in a fistula of Wharton's duct. AB - A 30-year-old woman developed a fistula of the Wharton's duct following excision of the submandibular gland. The contribution of conventional fistulography, MRI and MRI fistulography in detecting the exact extent of the fistula is discussed. PMID- 10808231 TI - 2nd European Meeting On Pulse Wave AnalysisThe Royal Society of Medicine London, UK. Monday 4th October 1999. PMID- 10808230 TI - Publisher's announcement. PMID- 10808232 TI - Viagra and its use in cardiovascular disease. PMID- 10808233 TI - Risk of premature birth in multifetal pregnancy. AB - The risk of preterm delivery (< 37 weeks of gestation) is approximately nine times higher in women with multifetal pregnancies than in women with singleton pregnancies. However, it is possible that the risk will vary according to gestational week. To assess the risk of premature birth within 1 week by gestational age among multifetal pregnancies and compare the estimated risk with that of singleton pregnancies, we analyzed 6,036,475 infants born in singleton pregnancies and 90,887 infants born in multifetal pregnancies in Japan (> or =22 weeks) over the 5-year period 1989-1993. An estimate of the risk of birth within 1 week at gestational week n was obtained by dividing the number of infants delivered at gestational week n by the number of infants delivered at or beyond gestational week n. The risk at 22 weeks was 0.9 per 1000 fetuses for singleton pregnancies and 5.0 per 1000 for multifetal pregnancies. The risk remained relatively stable until 27 weeks of gestation, then sharply increased toward 36 weeks of gestation in both singleton and multifetal pregnancies. The odds ratio for birth within 1 week for fetuses of multifetal pregnancies compared with fetuses of singleton pregnancies was 5.9 (95% CI, 5.4-6.5) at 22 weeks of gestation, increasing gradually with increasing gestational age until 33 weeks of gestation (13.7; 95% CI, 13.1-14.2) but declining thereafter to 8.8 (95% CI, 8.6 8.9) at 36 weeks of gestation. Results of data analysis for each year of the 5 year period did not differ substantially. PMID- 10808234 TI - Ultrasound examination of neonatal hip: correlation of twin pregnancy and congenital dysplasia. AB - Twin pregnancy is considered to be a risk factor for congenital dysplasia of the hip. From 1987 until 1996, the hips of 4476 (2260 male, 2216 female) newborn babies were examined by ultrasound according to Graf's technique and classification in our hospital. In this study, we compare the results of twins and singletons for this risk factor. Of the newborns, 97 (2.2%) were twins (40 male, 57 female); 39 pairs of twins (10 male/male, 19 female/female, 10 male/female) and 19 individual twins (6 male, 13 female) were investigated. Hips of type Ia, Ib and IIa (alpha > or = 55 degrees) are not pathologic; hips of type IIa (alpha <55 degrees ) need an early control examination; and hips of type IIc, D, IIIa, IIIb and IV require therapy. Types Ia, Ib, and IIa (alpha > or =55 degrees ) were found in 4207 (94.0%) of all newborns, in 4112 (93.9%) of the singletons, and in 95 (97.9%) of the twins. Early control examination and/or therapy (indicated for types IIa (alpha <55 degrees ), IIc, D, IIIa, IIIb, and IV) were necessary in 269 (6.0%) of all cases, in 267 (6.1%) of singletons and two (2.1%) of twins. Twins with additional factors such as breech position birth, hip dysplasia in the family or premature birth did not show the types of hip IIa (alpha <55 degrees ), IIc, D, IIIa, IIIb, IV. We did find these hips in two (3.5%) of the female twins, but not at all in the male twins. Statistically, twins with or without other risk factors that are known before birth did not show significantly more of type hip IIa (alpha <55 degrees ), IIc, D, IIIa, IIIb, IV (P>0.05). PMID- 10808235 TI - Dizygotic to monozygotic twinning ratio at The Royal Women's Hospital, Melbourne 1947-1997, compared with Australian national twinning incidence. AB - The incidence of dizygotic (Dz) twinning can be used as an index of natural human fertility. A retrospective study was done at The Royal Women's Hospital, Australia, to see whether the dizygotic to monozygotic (Mz) twinning ratio from one hospital can accurately reflect the national incidence of Dz twinning. The yearly twinning incidence from 1947-1997 was expressed as a Dz:Mz ratio, standardised for maternal age and plotted against previously published national statistics. The proportion of mothers born in Asia (of both singleton and multiples) between 1983-1997 was analysed to see whether different racial mixes might influence twinning trends. There were 5275 twins born of known sex and maternal age between 1947-1997. The age-standardised Dz:Mz ratio increased non significantly from 1.39 in 1947 to 2.29 in 1953 (P = 0.08), underwent a significant decline to 0.73 in 1977, then remained stable until 1997 (P>0.05). The same trends were also apparent when the data was pooled into 2-year groups with the increase from 1947/48-1953/54 becoming highly significant (P<0.009). These trends observed in the hospital-based data were in close agreement with those found in the national statistics, with the exception of a rise in 1977-1982 only reflected in the Australia-wide data. In 1993, 2.6% of mothers were born in Asian countries; by 1997, this had risen to 10.6%. We found that the Dz:Mz ratio from one hospital closely reflects national twinning trends. Prospective studies must account for race, and would need around 200-300 twin pairs per year to minimise fluctuations of the ratio. PMID- 10808236 TI - Happy families: a twin study of humour. AB - The objective of this study was to estimate how much of an individual's appreciation of humour is influenced by genetic factors, the shared environment or the individual's unique environment. A population-based classical twin study of 127 pairs of female twins (71 monozygous (MZ) and 56 dizygous (DZ) pairs) aged 20-75 from the St Thomas' UK Adult Twin Registry elicited responses to five 'Far Side' Larson cartoons on a scale of 0-10. Within both MZ and DZ twin pairs, the tetrachoric correlations of responses to all five cartoons were significantly greater than zero. Furthermore, the correlations for MZ and DZ twins were of similar magnitude and in some cases the DZ correlation was greater than that of the MZ twins. This pattern of correlations suggests that shared environment rather then genetic effects contributes to cartoon appreciation. Multivariate model-fitting confirmed that these data were best explained by a model that allowed for the contribution of the shared environment and random environmental factors, but not genetic effects. However, there did not appear to be a general humour factor underlying responses to all five cartoons and no effect of age was seen. The shared environment, rather than genetic factors, explains the familial aggregation of humour appreciation as assessed by the specific 'off the wall' cognitive type of cartoons used in this study. PMID- 10808237 TI - Aetiology of teenage childbearing: reasons for familial effects. AB - The aims of the present study were to evaluate the contribution of the genetic and environmental factors to the risk of teenage childbearing, and to study whether life style, socio-economic conditions, and personality traits could explain possible familial effects. We linked two population-based registers: the Swedish Twin Register and the Swedish Medical Birth Register. The study covers female twin pairs born between 1953 and 1958, having their first infant before the age of 30 years (n = 1885). In order to separate familial effects from other environmental influences, and genetic effects from shared environmental effects, only complete twin pairs with known zygosity were included, in all 260 monozygotic and 370 dizygotic twin pairs. We used quantitative genetic analyses to evaluate the importance of genetic and environmental effects for liability to teenage childbearing. Logistic regression analyses were used to estimate the effects of life style, socio-economic situation, and personality on the probability of teenage childbearing, and to study whether psychosocial factors could explain possible familial effects. Fifty-nine percent (0-76%) of the variance in being a teenage mother was attributable to heritable factors; 0% (0 49%) was due to shared environmental factors; and 41% (23-67%) was explained by non-shared environmental factors. Thus, the data were consistent with the hypothesis that the familial aggregation of teenage childbearing is completely explained by genetic factors, although the alternative hypothesis that familial aggregation is entirely explained by shared environmental factors cannot be ruled out. Significant effects of smoking habits, housing conditions, and educational level were found in relation to liability to teenage childbearing. However, the familial effects on risk of teenage childbearing were not mediated through similarities in life style and socio-economic factors. When studying risk factors for teenage childbearing, it is recommended to include life style and socio economic variables as well as information about family history of teenage childbearing. Twin Research (2000) 3, 23-27. PMID- 10808238 TI - Lifetime prevalence of mood and anxiety disorders in twin pairs discordant for schizophrenia. AB - There have been long questions about the relationship of schizophrenia to other mental disorders. Lifetime DSM-III-R diagnoses of mood and anxiety disorders in twins with clinically diagnosed schizophrenia (n = 24) and their non-affected co twins (n = 24) were compared with twins from pairs without schizophrenia (n = 3327) using a sample from the Vietnam Era Twin Registry. Schizophrenic probands had significantly elevated rates of all included disorders (bipolar disorder, major depression, dysthymia, generalized anxiety disorder, panic disorder, and PTSD) compared with controls (P<0.01). The odd ratios comparing co-twins of schizophrenic probands with controls was greater than three for every disorder, but did not attain statistical significance. A similar pattern was observed when analyses were restricted to only monozygotic twins (n = 12). Consistent with other studies, schizophrenics appeared to have higher rates of a range of mental disorders. Our results suggest that schizophrenia per se represents a risk factor for other psychiatric disorders, but the absence of significantly elevated risk among non-schizophrenic co-twins suggested that family environmental and/or genetic factors that contribute to risk of schizophrenia do not increase the risk of mood and anxiety disorders to the same extent that the risk of these other disorders is increased by the presence of schizophrenia. PMID- 10808239 TI - Representativeness of a roster of volunteer North American twins with chronic disease. AB - To identify large numbers of twins affected by chronic disease as potential subjects for studies of environmental and genetic chronic disease determinants, we advertised for affected twins over the period 1980-91 in newspapers across North America. Responses were received from 17 245 twin pairs in which cases of cancer or other chronic disease had occurred. To assess the representativeness of affected twins identified by advertising, we evaluated the pattern of reporting, compared the cases identified to the number of cases estimated to be prevalent among all North American twins, compared the cases to population-based singleton case series, compared the healthy co-twins to population-based samples of healthy persons, assessed the impact on ascertainment of opinions about disease causation, compared the pattern of prospective to retrospective ascertainment of disease in the originally unaffected co-twins of cases, and compared the results of the prospective ascertainment of disease in co-twins to comparable published estimates. Youth, gender, zygosity, education, and disease concordance were found to be overall determinants of ascertainment. Disease-discordant DZ twins appeared to be modestly underascertained. While somewhat better educated, both concordant and discordant pairs were judged to be reasonably representative of affected non Hispanic white North American twin pairs of comparable status, ie of comparable age, sex, race, and zygosity. If interpreted with caution, the concordance patterns of such twins can be used to generate genetic hypotheses, but should not be the basis of definitive heritability analyses. We conclude that advertising offers a method of identifying pairs of twins that can serve as subjects for studies designed to identify disease determinants. PMID- 10808240 TI - Genetic and environmental influences on body fat distribution, fasting insulin levels and CVD: are the influences shared? AB - Central body fat distribution has been shown to be related to hyperinsulinemia, insulin resistance, hypertriglyceridemia, and atherosclerosis to a greater degree than general obesity. There are known to be both genetic and environmental effects on all components of this clustering. Whether these genetic effects are due to one set of genes in common to the components or whether genetic influences on insulin resistance and/or general/abdominal fatness 'turn on' other genes that affect other components of the syndrome is not clear. We analyzed data from the Swedish Adoption/Twin Study of Aging (60% female; monozygotic = 116, dizygotic = 202; average age 65 years) to determine whether there were genetic and/or environmental factors shared among general body fat distribution, abdominal body fat distribution, fasting insulin levels and cardiovascular disease. We found additive genetic effects in males to be significantly different from those in females with genetic effects accounting for variance in waist-hip ratio (males = 28%; females = 49%), body mass index (males = 58%; females = 73%), fasting insulin levels (FI) (males = 27%; females = 49%), and cardiovascular disease (CVD) (males = 18%; females = 37%). There were also shared genetic and environmental effects among all the variables except CVD, but a majority of the genetic variance for these measures was trait specific. PMID- 10808241 TI - Genetic nature of individual frailty: comparison of two approaches. AB - The traditional frailty models used in genetic analysis of bivariate survival data assume that individual frailty (and longevity) is influenced by thousands of genes, and that the contribution of each separate gene is small. This assumption, however, does not have a solid biological basis. It may just happen that one or a small number of genes makes a major contribution to determining the human life span. To answer the questions about the nature of the genetic influence on life span using survival data, models are needed that specify the influence of major genes on individual frailty and longevity. The goal of this paper is to test the nature of genetic influences on individual frailty and longevity using survival data on Danish twins. We use a new bivariate survival model with one major gene influencing life span to analyse survival data on MZ (monozygotic) and DZ (dizygotic) twins. The analysis shows that two radically different classes of model provide an equally good fit to the data. However, the asymptotic behaviour of some conditional statistics is different in models from different classes. Because of the limited sample size of bivariate survival data we cannot draw reliable conclusions about the nature of genetic effects on life span. Additional information about tails of bivariate distribution or risk factors may help to solve this problem. PMID- 10808243 TI - Twin and Triplet Psychology: A Professional Guide to Working with Multiples. PMID- 10808242 TI - Robert Derom MD PHD FRCOG. PMID- 10808244 TI - Entwined Lives: Twins and what they tell us about Human Behavior. PMID- 10808246 TI - beta-glucuronidase activity is elevated in the rat harderian gland after bromocriptine but decreased after cyproterone acetate treatment or castration. Postcastrational effects on the ultrastructure of the gland. AB - OBJECTIVE: To investigate the effect of castration, bromocriptine and cyproterone acetate treatment on lysosomal hydrolytic enzymes and ultrastructure of the rat Harderian gland. METHODS: Groups of rats were subjected to the treatment by bromocriptine, cyproterone acetate and castration. Harderian glands were dissected from each experimental animal cut into small pieces and immediately stored in liquid nitrogen. Only those from castrated animals were fixed in Karnovsky fluid and processed for electron microscopy. The activity of acid phosphatase and ss-glucuronidase were estimated by kits from Sigma (St. Louis, Mo, USA). Semi-thin sections were stained with 1-% toluidine blue and ultra-thin sections with uranyl acetate and counter-stained with lead citrate. The examination was performed by Joel JEM-1200 EX II electron microscope. RESULTS: The treatment of male rats with the prolactin release inhibitor bromocriptine induced a significant increase in ss-glucuronidase activity. On the contrary, such activity was significantly decreased after treatment with the testosterone receptor antagonist cyproterone acetate as well as after castration. However, these treatments did not alter the activity of lysosomal acid phosphatase. Castration induced dramatic changes mostly in type A cells such as the appearance of lipid vacuoles with irregular forms and the predomination of smooth endoplasmic reticulum (SER) throughout the cytoplasm. The most dramatic postcastrational change was the degeneration of the mitochondria. These changes in type A cells might be due to the uneven distribution of the testosterone receptors in the rat Harderian gland, which are more numerous in type A cells. CONCLUSIONS: the gland physiology is responsive to alteration in circulating prolactin and testosterone levels. PMID- 10808247 TI - Comparison of antagonistic properties of substance P analogs, spantide I, II and III, on evoked tongue jerks in rats. AB - OBJECTIVE: To study and evaluate the effects of perfusion through cerebral ventricles with substance P (SP) and its analogs: spantide I, II and III on evoked tongue jerks (ETJ) in male rats. METHODS: During the perfusion, stimulation of the tooth pulp caused retractive movements of the stretched tongue, the amplitudes of which were recorded. The mean amplitudes of evoked tongue jerks (ETJ) recorded during each 10 min. period of perfusion with McIlwain Rodnight's solution and solutions containing peptides were compared. RESULTS: Perfusion of cerebral ventricles with SP caused a significant increase in the mean amplitude of evoked tongue jerks. Spantide I caused a complete respiratory arrest in all of the examined animals, so its effect on the trigemino-hypoglossal reflex could not have been tested. Spantide II, in the first two minutes, induced a transient significant decrease in ETJ amplitude, followed by an increase in ETJ in the next 8 min. SP perfused after spantide II caused a further significant increase in ETJ, as compared with control. Perfusion of cerebral ventricles with spantide III caused a significant, dose-dependent decrease in ETJ. SP perfused after spantide III caused a smaller increase in ETJ than it was observed without spantide III. CONCLUSION: Spantide III was found to be a strong antagonist of SP in trigemino-hypoglossal reflex. PMID- 10808248 TI - Huerthle cell neoplasms of the thyroid: predicting malignant potential. AB - OBJECTIVE: Assessment of malignancy criteria in Huerthle cell neoplasm. METHODS: This study intends to review retrospectively the patients who were operated for Huerthle cell neoplasia at Gazi University, Department of General Surgery between January 1986 and October 1999. Pathological specimens from 63 patients (20 males and 43 females) were investigated in this study, 48 of which revealed Huerthle cell adenoma and 15 revealed Huerthle cell carcinoma. The mean age of the patients with Huerthle cell adenoma was 40.7+/-1.59 yr while it was 51.3+/-1.83 yr in patients with Huerthle cell carcinoma. Mann-Whitney U and Chi-square tests were used for statistical analysis. RESULTS: . Fifty-two of the 63 patients had fine needle aspiration (FNA) biopsy prior to operation, 49 of those were reported to have suspected Huerthle cell neoplasia (HCN) and three had suspected Huerthle cell carcinoma (HCC). The sensitivity of FNA for HCN was 20 %, specificity was 100 %, positive predictive value was 100 % and negative predictive value was 76 %. For all patients, peroperative frozen section (FS) biopsy was examined. Fifty nine of the FS specimens revealed HCN and four revealed HCC. The sensitivity, specificity, positive predictive value and negative predictive value of FS biopsy were 27 %, 79 %, 28.5 % and 77.5 %, respectively. In this retrospective study, there was a statistically significant correlation between malignancy and the size of the tumor (P<0.05) according to Chi-square test, and also a statistically significant correlation between malignancy and the age of the patient (P<0.05) according to Mann-Whitney U test. CONCLUSIONS: In cases where FS and FNA biopsies cannot adequately define the benign or malignant behaviour of the tumor, the age of the patient and the diameter of the tumor must be taken into consideration for accurate surgical strategy. Particularly for 50 year-old and elderly, incidence of malignancy is statistically significant without considering sex of the patient. PMID- 10808249 TI - Radioimmunoassay for hypocretin-2. AB - OBJECTIVE: To develop radioimmunoassay for hypocretin-2 (Hcrt-2). And search for its presence in certain rat tissues. METHODS: Anti-Hcrt-2 serum has been raised in New Zealand white rabbits immunized with a conjugate of synthetic Hcrt-2 with bovine serum albumin. Radioiodination of Hcrt-2 was performed by chloramine T method, followed by purification of radoiodinated material on Sephadex G-25 column. RESULTS: The obtained antibody did not cross react with hypocretin-2, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. The assay was performed with a double antibody system. Hcrt-2 was extracted from the tissues with acid acetone. The dilution curve of acid acetone extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue Hcrt-2 was about 85 % and the intra-assay and inter-assay variation were 5.6 % and 8.0 %, respectively. Hcrt-2 was found in the hypothalamus, cerebrum, brain stem and testes. CONCLUSIONS: The obtained data suggest that the assay system developed is suitable to measure Hcrt-2 in tissues and that Hcrt-2 is mainly found in the hypothalamus. PMID- 10808250 TI - The use of GH as pharmacological agent (minireview). AB - Growth hormone (GH) has been traditionally used a drug for substitution therapy of GH deficiency, but since the wide availability of biosynthetic GH it has also been used as a pharmacological agent. In this report the presently available data are reviewed. PMID- 10808251 TI - Children born with intra-uterine growth retardation (IUGR) or small for gestational age (SGA): long term growth and metabolic consequences. AB - Intra Uterine Growth Retardation (IUGR) refers to insufficient fetal growth diagnosed either by two direct intrauterine growth assessment (ultra-sonography) or when the fetus or newborn length (height) is less than two standard deviations (or third percentile) below the mean for gestational age (NIKLASSON et. al. 1991; DE ZEGLER 1997). When the fetus or newborn body size (wight or length) is insufficient for gestational age, that is less than 2 standard deviations below the mean (or third percentile) for gestation during the situation is referred to as Small for Gestational Age (SGA). Since both fetal weight and length (height) gains are closely related, there is much overlap between SGA and IUGR. The proportion of newborn with normal birth weight and overlap between SGA and IUGR, isolated low birth weight, isolate low birth length and combined low birth weight and length is presented in Tab. 1, according to the most recent series reported nb NIKLASSON (1991). SGA/IUGR is a public health problem, since 2.5-3.0 % of newborns are affected by definition, and 8-10 % of them do not catch up postnatally, presenting with a persistent severe height deficiency, developmental difficulties and poor outcome (UNDERWOOD 1991; SIEGEL et al. 1991; Albertson WICKLAND et. al. 1993; LAKEMAN et al. 1994). PMID- 10808252 TI - How BCG led to the discovery of immunostimulatory DNA. AB - The concept of immunostimulatory DNA was borne in a long series of studies on BCG mediated tumor resistance. DNA purified from BCG inhibited the growth of various syngeneic animal tumors, augmented NK cell activity and induced IFN-alpha/beta and -gamma from mouse spleen cells and human PBL. Extending the lines of study, we found two biologically remarkable facts that (i) DNAs from invertebrates, but not from vertebrates and plants, showed the above-mentioned biologic activities, and (ii) the activities were completely dependent on particular base sequences having CpG motifs but in a senseless manner. Details of those early studies carried out mainly in the 1980's have been reviewed in the first part of this paper. In the middle part of this review, the results of toxicity and pharmacology studies and clinical trials of BCG- DNA, performed by other groups in Japan in the late 1980's, were introduced. Since a large amount of DNA had never been administered repeatedly into experimental animals or human, those experiences obtained seem to be worthwhile to introduce. Research interests of immunostimulatory DNA were galvanized in 1995 by the report of Krieg et al. showing murine B cell activation with bacterial DNA containing CpG motifs. Within a short period of time, a huge number of papers have been published in this field, and the study has expanded rapidly and largely. Now, it includes a number of research fields, for example, host-defense mechanisms against infection, allergy, autoimmune diseases, cytokine networks, plasmid vaccination, and therapeutic application of certain diseases. This paper reviewed briefly recent advances of immunostimulatory DNA research. The response of higher animals against immunostimulatory DNA must be the most primitive but important mechanism for self-nonself discrimination against foreign DNA. By utilizing immunostimulatory DNA or controlling this primitive response, it seems possible to offer many beneficial means to human health. For instance, more potent peptide or plasmid- vaccines could be developed by the use of immunostimulatory DNA. On the other hand, many study results suggest that immunostimulatory DNA works either beneficially or harmfully for the hosts. We assume that further extensive and careful studies are required. PMID- 10808253 TI - Enterovirus 71 from fatal and nonfatal cases of hand, foot and mouth disease epidemics in Malaysia, Japan and Taiwan in 1997-1998. AB - Enterovirus 71 (EV71), one of the major causative agents for hand, foot and mouth disease (HFMD), is sometimes associated with severe central nervous system diseases. In 1997, in Malaysia and Japan, and in 1998 in Taiwan, there were HFMD epidemics involving sudden deaths among young children, and EV71 was isolated from the HFMD patients, including the fatal cases. The nucleotide sequences of each EV71 isolate were determined and compared by phylogenetical analysis. EV71 strains from previously reported epidemics belonged to genotype A-1, while those from recent epidemics could be divided into two genotypes, A-2 and B. In Malaysia, genotype A-2 was more prevalent, while in Japan and Taiwan, B genotype was more prevalent. Two isolates from fatal cases in Malaysia and one isolate from a fatal case in Japan were genotype A-2. However, all isolates from three fatal cases in Taiwan belonged to genotype B. The severity of the HFMD did not link directly to certain genotypes of EV71. PMID- 10808254 TI - Enterotoxigenicity of Klebsiella pneumoniae associated with childhood gastroenteritis in Madras, India. AB - Klebsiella pneumoniae was isolated as the predominant growth from 12 of 100 children under 3 years of age suffering from acute diarrhea. Of the 12 isolates, SA1, SA2, SA4, SA5, SA6 produced a secretogenic response in the ligated rabbit ileal loop, and one isolate, SA3, induced a diarrheagenic response in suckling mice. Two isolates, SA7 and SA8, were diarrheagenic in both assays. Strains SA9, SA10, SA11, and SA12 were found to be non-enterotoxigenic. These isolates belonged to serotypes K6, 16, 25, 30, 39, 46, 49, 53, 66, and 81. All eight enterotoxigenic strains were resistant to ampicillin, streptomycin, ceftazidime, cefuroxime, and cotrimoxazole. Only quinolones such as ciprofloxacin and norfloxacin appear to be effective against enterotoxigenic K. pneumoniae. PMID- 10808255 TI - Detection and identification of adenovirus from ophthalmological specimens by virus isolation and PCR. PMID- 10808256 TI - Surveillance of poliovirus-isolates in Japan, 1998. PMID- 10808257 TI - Malaria infections in Japan. PMID- 10808258 TI - Prevalence of drug resistance-related mutations among HIV-1s in Japan. PMID- 10808259 TI - The impact of ventilation systems on worker exposures during advanced composite material and fiberglass repair operations on military aircraft. PMID- 10808260 TI - Control of scrap paper baler crushing hazards. National Institute for Occupational Safety and Health. PMID- 10808261 TI - Carbon monoxide exposure from a propane-fired concrete cutter. PMID- 10808262 TI - Exposures to asbestos arising from bandsawing gasket material. AB - A simulation of bandsawing sheet asbestos gasket material was performed as part of a retrospective exposure evaluation undertaken to assist in determining causation of a case of mesothelioma. The work was performed by bandsawing a chrysotile asbestos (80%)/neoprene gasket sheet with a conventional 16-inch woodworking bandsaw inside a chamber. Measurements of airborne asbestos were made using conventional area and personal sampling methods, with analysis of collected samples by transmission electron microscopy (TEM) and phase contrast microscopy (PCM). These were supplemented by qualitative scanning electron microscopy (SEM) examinations of some of the airborne particles collected on the filters. In contrast with findings from studies examining manual handling (installation and removal) of gaskets, airborne asbestos concentrations from this operation were found to be well above current Occupational Safety and Health Administration (OSHA) permissible exposure limit (PEL) (eight-hour time-weighted average [TWA]) and excursion limit (30-minute) standards. Although some "encapsulation" effect of the neoprene matrix was seen on the particles in the airborne dust, unencapsulated individual fiber bundles were also seen. Suggestions for the implications of the work are given. In summary, the airborne asbestos concentrations arising from this work were quite high, and point to the need for careful observation of common sense precautions when manipulation of asbestos containing materials (even those believed to have limited emissions potential) may involved machining operations. PMID- 10808263 TI - The verification of hazardous ingredients disclosures in selected material safety data sheets. AB - Under the provisions of the Workplace Hazardous Materials Information System, workers in Canada must be provided with accurate and comprehensive Material Safety Data Sheets (MSDSs) describing controlled products used in the workplace. As part of an ongoing auditing project, the MSDSs of some controlled products in use under federal jurisdiction were assessed for accuracy and completeness of their ingredient disclosures. Chemical analyses of samples using gas chromatography-mass spectrometry, infrared spectrophotometry, X-ray fluorescence, and wet methods, were performed to verify the ingredient disclosures in accompanying MSDSs. In this article, analytical processes and results are presented for three cases in which MSDS ingredient disclosures were incomplete. The products included a synthetic lubricant used in a mining operation, a detergent concentrate used for aircraft cleaning, and an epoxy reducer used in aircraft maintenance. In each case, undisclosed hazardous ingredients were detected at concentrations which required their disclosure. In at least one of these cases, the information provided in other sections of the MSDS failed to adequately describe the hazards and required protective measures for the composition discovered. Because the results suggest circumstances in which the inaccurate MSDS could act as a mechanism for workplace injury, compliance measures including employer, inspector, and user education, improved MSDS writer qualifications, and the incorporation of chemical analysis in active auditing programs are recommended. PMID- 10808264 TI - Effect of varying the combustion parameters on the emissions of carbon monoxide and nitrogen oxides in the exhaust gases from propane-fueled vehicles. AB - Propane-fueled forklifts are one source of carbon monoxide (CO) contamination of workplace air. The previous study carried out by the Quebec Occupational Health and Safety Research Institute dealt with worker exposure to CO during forklift use in buildings. It recommends that exhaust gas emissions be kept below a 1 percent concentration. However, this control has not produced a significant reduction in worker exposure to CO, when factors (ventilation, type of work tasks, and management of vehicle fleet) specific to companies are taken into account. Consequently, a reduction in CO emissions below the threshold of 0.3 percent should be considered. The experience acquired with propane-fueled ice resurfacers can be used to determine the effect of combustion parameters on exhaust gas emissions. It is known that a reduction in CO emissions from ice resurfacers resulted in the appearance of nitrogen oxides (NOx) and eventually in nitrogen dioxide (NO2) poisoning. Few publications present NOx results in relation to the CO measured in the exhaust gases of propane-fueled vehicles. The objective of this study is to define the level to which CO emissions can be reduced without increasing NOx concentrations. This real-situation study quantified the CO, NO, and NOx in the exhaust gases of a fleet of propane-fueled forklifts in relation to the mixture ratio. The results show the impact of the motor speed and mixture ratio on the CO, NO, and NO2 concentrations. They confirm an increase in NOx concentrations when CO concentrations are reduced. They also show that proper maintenance of forklifts combined with optimal adjustments can reduce CO and NOx emissions. The study proposes a compromise between CO and NOx emissions by taking into account worker health and safety as well as vehicle performance. Monitoring must be done to control air quality in work areas and worker exposure to CO and NO2. A forklift preventive maintenance program and general building ventilation are the favored and recommended means of control. PMID- 10808265 TI - Work-related cumulative trauma disorders and interpreters for the deaf. AB - With the passage of the Americans with Disabilities Act (ADA), increasing numbers of interpreters for the deaf are being employed in education, industry, or other settings. Professional interpreters are at risk for developing cumulative trauma disorders due to the nature of their work which involves rapid repetitive movements of the arms and hands. One hundred forty-five interpreters for the deaf responded to a survey questionnaire designed to identify their work experiences and development of physical pain/discomfort related to work. Incidence and duration of pain/discomfort is positively correlated with hours worked and time of onset. Regardless of age, training, or work experience, 119 (82%) of these respondents experienced disabling pain/discomfort during and following work. Thirty-nine (33%) of the 119 respondents indicated onset of pain or discomfort in the wrist and hand. These data suggest that interpreting for the deaf may result in debilitating pain/discomfort in hands, arms, shoulders, and back if the interpreter is not appropriately prepared to compensate for physical and attentional stresses. Implications include the need to provide periods of rest while working and to provide training for students learning to interpret for the deaf to minimize physical stress while working. PMID- 10808266 TI - Exposure assessment in the occupational setting. AB - Exposure assessment, the first step in risk assessment, has traditionally been performed for a variety of purposes. These include compliance determinations; management of specific programs that are implemented by comparison with an occupational exposure limit (such as medical surveillance, training, and respiratory protection programs); task/source investigations for determination of exposure control strategies; epidemiologic studies; worker compensation/toxic tort cases; health complaint or problem investigations; risk assessment and management; and evaluation of future changes in the workplace (e.g., introduction of a new chemical). Each purpose requires slightly different approaches, but there are also many similarities. The goal of this paper is to identify a general approach to assessing exposures that can be used for all purposes with only slight modifications. Five components of exposure assessments are identified: collection of data, identification of the hazard, selection of exposure metrics, definition of exposure groups and estimation of the exposures. The characteristics of these components for each type of assessment are discussed. From this review, it is clear that there is substantial overlap across the types of assessment. A single exposure assessment program is suggested that encompasses all the needs of these assessments and incorporates assessment of exposures for an entire workforce at a site at minimal cost by using prediction models and validation with measurements. PMID- 10808267 TI - An assessment of lead controls for torch cutting and rivet removal on steel structures. AB - The use of engineering and work practice controls to protect workers from lead containing dusts and fumes generated during rehabilitation of steel structures is mandated by the Occupational Safety and Health Administration (OSHA) Lead in Construction Standard (1993). Because the implementation and assessment of controls can be problematic in the rugged and dynamic construction environment, industrial hygienists should understand the effectiveness and limitations of controls adopted. The present investigation assesses the efficacy of two controls to reduce lead exposure: paint removal prior to oxy-acetylene torch cutting of steel, and encapsulation of rivets prior to their removal. A task-based exposure assessment approach was used to evaluate these tasks at three sites. Exposures at one site without controls were compared to exposures at sites with controls. Comparison of the results via an analysis of variance (0.05 significance level) indicates that, for torch cutting, exposures at the control site were not significantly different from those at an uncontrolled site (p = 0.14). The results for rivet busting show no significant differences in exposures at the control site compared to the uncontrolled site (p = 0.08). Results are also presented from two control sites where work was done in enclosed spaces. Two main difficulties in applying the controls are explored: technical and managerial. Technical problems during torch cutting included the penetration of paint into the steel profile and the configuration of the structures. For rivet busting, working within an enclosure was an important factor. Management problems arose both from a lack of coordination among different contractors, and from a failure to provide day-to-day guidance and assessment of the control. Important components of a program to implement controls are preplanning and coordination of control implementation, frequent testing of control efficacy, and a method for timely intervention to correct deficiencies. PMID- 10808268 TI - Cell proliferation rates and fibronectin arrangement as parameters for biocompatibility evaluation of dental metal alloys in vitro. AB - A short-term (72-96 hours) biocompatibility evaluation in vitro of four single phase dental metal alloys was conducted by determining cell proliferation rates correlated to the organization of the extracellular matrix protein fibronectin in human fibroblast cultures. Immunocytochemical methods were performed to detect both cell proliferation rates by 5-bromodeoxyuridine (BrdU) incorporation, and fibronectin arrangement, i.e., diffuse in the extracellular matrix, organized in fibrils or in focal adhesions. We showed that cell proliferation rates were related to fibronectin expression. In particular, a higher percentage of cells in the S-phase were related to a predominance of fibronectin organized both in fibrils and in focal adhesions. The alloy with the highest Au content seemed the most biocompatible among those tested, since it behaved in a very similar manner to the controls. On the contrary, fibroblasts exposed to the alloy with the highest percentage of Ag had the most different behavior as compared to the controls. We can assume that a correlation exists between fibronectin organization and the percentage of BrdU-positive cells and that these parameters are varying with the different metal composition of the alloys. The observation of fibronectin arrangement together with cell proliferation rates could be considered a useful tool to determine the biocompatibility of these biomaterials. PMID- 10808269 TI - Clinicopathologic study of odontogenic keratocysts in Singapore and Malaysia. AB - This was a retrospective study of odontogenic keratocysts in people from the Singapore-Malaysian region. The purpose of this study was to present the clinicopathologic features of odontogenic keratocysts in the Oriental population and to compare these data with those from other reported studies. Biopsy records from 1981 to 1992 of 61 cases of odontogenic keratocysts from patients in Malaysia and Singapore showed that 42.6% of patients were female and 57.4% of patients were male. Among patients with cysts, 75.4% were Chinese, 6.6% were Malays, 9.8% were Indians and 8.2% were other ethnic groups. The mean age of these patients was 26.98 +/- 15.38 years with a peak incidence occurring in the second to fourth decades. The location of the lesions was more often in the mandible (65.5%) than the maxilla (31.0%). There was a marked predilection for lesions to occur in the posterior mandible. Histologically, 90.2% of the cysts were lined with a para-keratinized stratified squamous epithelium while only 3.3% of the cysts were lined with orthokeratinized stratified squamous epithelium. Mixed para-keratinized and orthokeratinized epithelial linings were observed in 4 cases (6.5%). The cyst linings were mainly uninflamed (95.1%). Inflammation of the cyst wall was found in 42 cases (68.8%). Twelve (19.7%) cases contained keratin in the lumen. A satellite cyst was observed in only 6 cases (9.8%). In conclusion, most clinical and histological features seen in this study were similar to those found for Caucasians. The only clinical feature that was different was the peak age incidence, that ranged from the second to fourth decades, with an absence of a second peak. Odontogenic keratocysts presenting at the site of the dentigerous cyst were observed in 7 cases (11.5%). PMID- 10808270 TI - Oral recurrent aphthous ulcers/stomatitis: prevalence in Malaysia and an epidemiological update. AB - One of the objectives of this short communication was to add to the literature on the prevalence of oral recurrent aphthous stomatitis (ORAS). This research is based on a total of 11,697 randomly selected Malaysian subjects with an age range of 25-115 years and a mean age of 44.5 +/- 13.9 years who were examined for oral mucosal lesions (ORAS). The prevalence of ORAS detected during the oral examination (average point prevalence) was found to be 0.5% (64 subjects). ORAS formed 5.7% of all lesions detected during the survey. The average point prevalence of ORAS was highest in the indigenous people of Sabah and Sarawak (1.2%), followed by the Chinese (0.7%), the Malays (0.5%) and the Indians (0.1%). This difference was statistically significant (p < 0.001). A review of the English literature on the prevalence of ORAS revealed different prevalence types used by different researchers, namely average point prevalence (APP), self reported life-time prevalence (SLP) and self reported two-year prevalence (STP). The other objective of this paper was to present the different types of prevalence that have been reported in the literature and to discuss the usefulness of such prevalence types in relation to using epidemiology in deriving certain possible etiological associations. PMID- 10808271 TI - Molecular genetic study of the gutter shaped root (GSR) on mouse chromosome 17. AB - In this study, a genetic factor affecting the appearance of the gutter-shaped root (GSR) in mice was examined using two inbred strains of mice, C57L/J and C57BL/6J, and the genetic crosses of these strains. 120 Mit markers, which exist on mouse chromosome 17, have been investigated to select polymorphic markers between the two strains. Nine Mit markers with polymorphism between the two strains were obtained using polymerase chain reaction (PCR). To detect the individual genotype of 66 backcross mice, simple sequence length A polymorphism (SSLP) analysis was performed. From the results of individual genotyping using 9 informative markers, the highest linkage was determined at D17Mit260. Based on these findings, it was suggested that the gene causing the mouse GSR might be located on mouse chromosome 17 and nearby D17Mit260. PMID- 10808272 TI - A new assay system for detection of polymeric immunoglobulin A-polymeric immunoglobulin receptor binding. AB - A new system for detection of polymeric immunoglobulin A (pIgA)-polymeric immunoglobulin receptor (pIgR) binding was established. Cell lysates of a mouse pIgR cDNA transfectant, 2S9.1, were incubated with mouse polymeric immunoglobulin A (pIgA) or immunoglobulin G (IgG). The resulting immunocomplexes were precipitated with protein G-Sepharose and blotted with polyclonal anti-mouse pIgR antibody. The mouse pIgR molecule was specifically precipitated with pIgA, indicating successful detection of the pIgA-pIgR complex. Using this system, the role of N-glycosylation in pIgA-pIgR binding was examined. The pIgR molecule (molecular mass 100 kDa) after complete deglycosylation by tunicamycin treatment was still able to bind to pIgA, indicating that N-glycosylation of pIgR is not necessary for pIgA-pIgR binding. This novel system will be useful for detecting pIgA-pIgR complexes containing intact pIgR molecules. PMID- 10808273 TI - The effects of attachment factors on initial attachment of human periodontal ligament fibroblasts on different root surfaces: a light and scanning electron microscopic study. AB - This study was aimed to test the effect of fibronectin (FN), vitronectin (VN) and a fibronectin analog (fibronectin-like engineered protein) on the attachment of periodontal ligament cells to mechanically-treated and mechanically non-treated periodontally involved and non-diseased root surfaces in vitro. Periodontal ligament fibroblasts were incubated with a total of 44 periodontally diseased and non-diseased root slices which had been treated in the following manner: 1) FN applied to mechanically-treated and non-treated root slices, 2) VN applied to mechanically-treated and non-treated root slices, 3) FN-like engineered protein applied to mechanically-treated and non-treated root slices, and 4) mechanically treated and non-treated root slices. After the 1 hour incubation period in a humidified atmosphere of 95% air and 5% CO2 at 37 degrees C, the adherence of the fibroblasts was determined using light microscopy with an ocular grid system and orientation was evaluated by scanning electron microscopy. The results indicated that the number of attached cells to non-diseased cementum sites was significantly greater than the number of attached cells to diseased cementum sites for all of the groups (p < 0.05). Likewise, the number of attached cells to mechanically-treated diseased cementum sites was significantly greater than the number of attached cells to mechanically-non-treated diseased cementum sites (p < 0.05). Our findings suggest that these attachment factors cannot promote cell attachment to different cementum sites. PMID- 10808274 TI - Myoepithelioma arising from the buccal gland: histopathological and immunohistochemical studies. AB - A rare case of myoepithelioma of the buccal gland in a 54-year-old Japanese woman is reported. As the swelling exhibited a normal mucosal color and was relatively well defined, showing no ulcers, a benign salivary gland tumor was suspected upon clinical inspection. Microscopically, the parenchyma of the present case mainly consisted of plasmacytoid cells with round nuclei and eosinophilic cytoplasm, and partial spindle cells with eccentric nuclei. The stroma was composed of fibro hyalinized or myxoid connective tissue that separated from the parenchyma. Immunohistochemically, the cytoplasm of the plasmacytoid and spindle cells was moderately positive for vimentin and GFAP, whereas the buccal gland adjacent to the tumor was negative for these antibodies. S-100 protein reactivity is strong for both types tumor cells. Actin reactivity was negative for both types of tumor cells, notwithstanding the fact that myoepithelial cells of the buccal gland were positively stained. Anti-cytokeratin reactivity was weak for both types of tumor cells in portions of the plexiform and solid areas; nevertheless, the buccal glands were moderately positive. These results suggest that neoplasmic myoepithelial cells exhibit abnormal differentiation and modification. There have been only two published reports of myoepithelioma arising from the buccal gland in the literature to date. PMID- 10808275 TI - Case report of a pigmented dentigerous cyst and a review of the literature on pigmented odontogenic cysts. AB - This paper reports the first case of a dentigerous cyst containing melanin pigment and melanocytes in the lining epithelium, and the first case of a odontogenic cyst with macroscopically visible pigmentation in the cyst wall. The patient was a 29-year-old Japanese male with a cystic lesion in the left retromolar area of the mandible. Pathologic examination revealed the lesion to be a dentigerous cyst with or without mild surface keratinization, and numerous granules of melanin-pigment were distributed in the basal cells of the epithelial lining. Furthermore, dendritic melanocytes were scattered in the basal layer. Review of the literature revealed that pigmented odontogenic cysts are uncommon, and only 11 cases have been documented; eight were odontogenic keratocyst, one was a gingival cyst, one was a botryoid odontogenic cyst, and one was a lateral periodontal cyst. The possible origin of melanocytes in odontogenic lesions is discussed. PMID- 10808276 TI - [Evaluation of CT appearance of Mycobacterium avium complex infection--comparison with bronchiectasia]. AB - We compared computed tomographic pictures (CT) of primary pulmonary mycobacterium avium complex (MAC) infection with bronchiectasia (BE). These patients were examined during the period from 1988 to 1996. There were 51 patients of pulmonary MAC infection group (13 males and 38 females) with a mean age of 67.5 years and 27 patients of BE group (8 males and 19 females) with a mean age of 53.9 years. The results were as follows: 1. In both groups, bronchiectasis was most commonly found in the middle lobe and in the lingula. In pulmonary MAC infection group, 38 patients (74.5%) had bronchiectasis in the middle lobe, and 40 patients (78.4%) had them in the lingula. In BE group, 18 patients (66.7%) had them in the middle lobe and in the lingula respectively. In the lower lobe, bronchiectasis of BE group was observed more frequently. And in pulmonary MAC infection group, 15 patients (29.4%) had bronchiectasis in the central zone, 42 patients (82.4%) had them in the intermediate zone, and all 51 patients had them in the peripheral zone. On the other hand, in BE group, 18 patients (66.7%), 27 patients (100%) and 21 patients (77.8%) had them in the central zone, in the intermediate zone and in the peripheral zone respectively. Bronchiectasis of pulmonary MAC infection was observed in peripheral zone more frequently than that of BE group. 2. By the type of bronchiectasis, saccular bronchiectasis was observed more frequently in BE group than in pulmonary MAC infection group. 3. Nodules were seen in pulmonary MAC infection group significantly more than in BE group. They were seen in 96% of lobes with bronchiectasis, and 50% of lobes without bronchiectasis in pulmonary MAC infection group. It was thought that the bronchiectasis with nodules predominantly in the peripheral zone was a characteristic CT finding of primary pulmonary MAC infection. As this finding was different from that of BE, it is suggested that primary pulmonary MAC infection caused this type of bronchiectasis. PMID- 10808277 TI - [A clinical study on pulmonary tuberculosis in younger age groups]. AB - In 1997, the number of newly registered patients with pulmonary tuberculosis increased, compared with that in 1996, in Japan. The majority of the increase were occupied by elder patients 70 years of age or higher. But in younger group less than 30 years old, a reduction in the incidence of tuberculosis had been slowed down, until 1996. The purpose of this report is to elucidate the characteristics of these younger patients. 139 cases younger than 30 years of age, who were hospitalized in the tuberculous ward of IMCJ from April 1995 to March 1998, were investigated, and were compared with the control group (557 cases), 30-79 years old who were hospitalized during the same period. In the younger group, the proportion of women cases, discovered by health examination, foreigners, and contact with TB patients in the past was significantly higher than in the control group. But there were no difference between the both groups, concerning the proportion of those spending irregular life or living alone. The proportion of sputa smear negative cases was significantly higher in the younger group than in the control. For early diagnosis of TB among younger group, the application of bronchofiberscopy and nucleic acid diagnostic method, are encouraged. PMID- 10808278 TI - [The frequency profile of tuberculin skin testing among employees of a hospital]. AB - The frequency profile of tuberculin skin testing among employees of a hospital was studied. In the analysis, tuberculin skin testing was defined as "negative" if skin rash is measured < or = 9 mm, and as "positive" if skin rash is measured > or = 10 mm, and as "strongly positive" if skin rash is measured > or = 10 mm, with induration, and double skin rash or bulla. Boosting was defined as initial skin rash was measured < or = 9 mm, with second skin rash measured > or = 10 mm and at least 6 mm increase of rash than the initial one. 544 employees were tested initially, and 81 were retested. Initial tuberculin positivity was associated with older age (p < 0.001); but boosting was not associated with age and occupation. Comparing pharmacists with other groups, the adjusted odds ratio of an initial strongly positive tuberculin reaction as significantly higher, namely 2.8 (95% confidence interval 1.2-6.7), but that of a booster reaction was 4.8 (0.7-32.1) which was insignificant. PMID- 10808279 TI - [Tuberculin survey of university students and postgraduates in 1998]. AB - As no recent report has been made concerning the tuberculin survey of healthy young adults in Japan, we checked all the Tohoku University undergraduate and postgraduate students by tuberculin test for the assessment of TB-situation in 1998. The sample included 5,517 students and postgraduates (3,888 males and 1,629 females; 31% of all subjects to be tested) who were evaluated on the basis of the redness of the skin test. The average age of the subjects was 22.3 +/- 3.1 years old (ranging from 18 to 51). As a result of this survey, 5,032 (91.2%) were positive and 485 (8.8%) were negative, and the average diameter of redness was 28.5 +/- 19.2 mm. Non-reactors received an additional BCG vaccination. Subjects showing strongly-positive (20.1%) results were rechecked by physical examination and chest X-ray, and none was found to require treatment by anti-TB drugs. Our findings also demonstrated that the diameter of redness was larger in the group previously BCG-vaccinated than in the group who had not received BCG vaccination previously (p < 0.01). PMID- 10808280 TI - [Pulmonary infection caused by Mycobacterium gordonae in immunocompetent patient]. AB - We report a case of 67-year-old female immunocompetent patient admitted to our hospital because of hemoptysis. Computed tomography (CT) of the lung showed bronchiectasis in the right S5 and small nodules in the right S6 and left S5. The cultures of sputum and bronchial washing specimen repeatedly revealed acid-fast bacilli identified as Mycobacterium gordonae (M. gordonae) by DNA-DNA hybridization (DDH) method. Thus, she was diagnosed to be infected with M. gordonae. She was treated with isoniazid, rifampicin, ethambutol and streptomycin. After treatment, the cultures of sputum and bronchial washing specimen converted to negative, and the chest CT showed improvement of small nodules. M. gordonae is a nontuberculous mycobacterium of very low pathogenic potency. Recently there have been a few reports of infection by M. gordonae not only in immunocompromised patients but also in immunocompetent patients. These cases were considered to be sensitive to initial standard antimycobacterial therapy, therefore, it is important to examine for M. gordonae in cases suspected of nontuberculous mycobacterial infection. PMID- 10808281 TI - [Simultaneous operation for lung tumor and cardiovascular disease]. AB - Successful simultaneous operation for lung tumor and cardiovascular disease was performed in three cases. A 76-year-old man with stage I lung cancer and ischemic heart disease underwent a partial lobectomy following single coronary artery bypass grafting through a median sternotomy. A 62-year-old man with stage I suspected lung cancer and thoracic aortic aneurysm underwent a partial segmentectomy before aneurysmectomy and patch closure using vascular prosthesis through a left posterolateral thoracotomy. These two cases were performed under extracorporeal circulation. A 69-year-old man with bronchogenic carcinoma and abdominal aortic aneurysm underwent a left upper lobectomy with standard lympho node dissection following aneurysmectomy and grafting using vascular prosthesis. As a simultaneous procedure, limited operation for lung tumor, especially for stage I non-small cell lung cancer, is acceptable for cases in using extracorporeal circulation. On the other hand, except emergency ruptured cases of abdominal aortic aneurysm, standard radical operation for lung cancer as a simultaneous procedure is preferred for cases such as lung cancer accompanied with abdominal aortic aneurysm without extracorporeal circulation. PMID- 10808282 TI - [A case of heart surgery without homologous transfusion for complete atrioventricular septal defect associated with tetralogy of Fallot]. AB - We performed intracardiac repair in an 11-month-old infant, weighing 5.1 kg, with complete atrioventricular septal defect and tetralogy of Fallot, who had been on a respirator, preoperatively, because of congestive heart failure due to severe common atrioventricular valve regurtitation. The preoperative angiogram revealed a very narrow and hypoplastic left pulmonary artery. Autologous blood donation after induction of anesthesia and minimalization of the cardiopulmonary bypass circuit (priming volume: 230 ml) made it possible to perform open-heart surgery without homologous blood transfusion in this case. The perioperative hemodynamics and respiratory status were satisfactory. An angiogram taken 1 year after surgery showed a well-developed left pulmonary artery. PMID- 10808283 TI - [A surgical case of papillary muscle reconstruction and double-CABGs for mitral regurgitation due to ruptured papillary muscle after myocardial infarction]. AB - A 76-year-old female patient underwent papillary muscle reconstruction, and d CABGs a month after the onset of myocardial infarction. Posterior papillary muscle (PPM) was recognized to be ruptured partially, its stump was sewn to original PPM. An ePTFE suture (CV 5) was placed from anterior mitral leaflet to anterior papillary muscle in attempt to reinforce PPM-repair. Carpentier-Edwards ring was inserted and d-CABGs (LITA to LAD and GEA to RCA) were performed, simultaneously. Postoperative examination revealed no regurgitation and no stenosis of bypass grafts during follow-up period of 5 months after the procedure. Papillary muscle reconstruction was effective procedure for MR due to the ruptured papillary muscle. PMID- 10808284 TI - [A case of primary intrapulmonary thymoma: its entity and the problem of lymph node dissection]. AB - Primary intrapulmonary thymomas are defined as primary thymomas arising in an intrapulmonary location without an associated mediastinal component, and they are very rare. A total of 20 cases have been reported only sporadically in the English literature since 1951. We reported the case of 41-year-old woman who had a 3.5 x 3.0 x 3.0 cm lower right lobe mass with nodal metastasis that extended over the left atrium. We also summarized the clinicopathological features of a total of 21 cases and discussed the problems involved with diagnosis, pathogenesis and treatment. Knowledge of the biological behavior of primary intrapulmonary thymomas is limited because of their rarity. In particular, the issue of the need for lymph node dissection has not been adequately discussed. In this case, pathohistological examination revealed that the routes of lymphatic spread and the sites of noda metastases from primary intrapulmonary thymoma resemble those of primary lung cancer. Therefore, systematic mediastinal lymph node dissection according to the lymph node map for primary lung cancer should be recommended for malignant cases. PMID- 10808285 TI - [The relationship between the tumor uptake of gallium-67 and the effect of cisplatin-based preoperative chemotherapy or chemoradiotherapy for lung cancer]. AB - Cisplatin has been usually used as the chemotherapy for lung cancer presurgically, though it is still difficult to predict downstaging of tumor from this drug. Four patients who underwent surgery achieved pathologic complete remissions in response to preoperative cisplatin-based chemotherapy with or without radiation. All of the Ga-67 citrate images in the four patients showed markedly increased uptake in tumors. According to the literatures, both Ga-67 and cisplatin binds to transferrin in the blood and transfers into the cells through transferrin-receptors expressed on the cell surface. The mechanism for Ga-67 and cisplatin uptake into tumor cells were alike. Marked tumor uptake on Ga-67 scintigraphy suggested that cisplatin-based chemotherapy or chemoradiotherapy was efficacious. PMID- 10808286 TI - [Vein holder--a new device for coronary artery bypass grafting]. AB - A new saphenous vein graft holder for coronary artery bypass grafting was described. This graft holder is a modification of a tissue forceps, the tips of which were changed to hollow-beveled, spoon shape. A saphenous vein graft can be pinched between the tips of this holder securely and gently during anastomosis. It enables us to place precise stitches and to minimize handling, anastomosis time, and leakage. PMID- 10808287 TI - [Mitral valve regurgitation associated with severe pulmonary hypertension successfully treated with nitric oxide gas: report of a case]. AB - A 61-year-old woman with mitral valve regurgitation and pulmonary hypertension, was referred to us for surgical repair. Preoperative left ventriculography showed Sellers IV mitral valve regurgitation and high pulmonary arterial pressure: 103/31 (57) mmHg. The mitral valve was replaced with a phi 27 mm SJM mechanical valve. The postoperative residual pulmonary hypertension was successfully treated with nitric oxide gas, decreased significantly pulmonary arterial pressure, and the postoperative course was uneventful. Based on our experience, we think that nitric oxide gas may prove effective for residual pulmonary hypertension after a cardiac operation. PMID- 10808288 TI - [Coronary artery bypass grafting by warm blood cardioplegia and normothermic cardiopulmonary bypass]. AB - Warm blood cardioplegia and normothermic cardiopulmonary bypass (CPB) have been used in coronary artery bypass grafting (CABG). The method of myocardial protection was intermittent combined antegrade and retrograde warm blood cardioplegia with terminal warm blood cardioplegia. We performed elective CABG in 30 patients above the age of 70 years (elderly group). These patients were compared with 30 patients below 70 years who underwent elective CABG (young group). No significant differences were observed about the preoperative data between two groups. No significant differences were obtained in the postoperative cardiac function, cerebral or renal complication between two groups. Warm blood cardioplegia and normothermic CPB were not associated with adverse effects on postoperative recovery in elderly as well as young patients. We may conclude that warm blood cardioplegia with normothermic CPB is a safe procedure for CABG in elderly as well as young patients. PMID- 10808289 TI - [Catastrophic pulmonary vasoconstriction associated with protamine reversal of heparin]. AB - Administration of protamine intravenously to neutralize the anticoagulant effects of heparin may be associated with hypotension. These adverse cardiovascular responses to protamine fall into three distinct types: transient hypotension related to rapid drug administration; anaphylactic responses: and catastrophic pulmonary vasoconstriction. We recently observed three patients who had catastrophic pulmonary vasoconstriction, which were treated with the assist circulation and intraaortic balloon pumping. The two patients were survived except one who was dead due to the cerebral ischemic damage which was introduced by delayed establishment of circulatory assist device. We conclude that meticulous longstanding observation of the hemodynamic condition is important after administration of protamine under the unremoved cannulations. PMID- 10808290 TI - [A new method of reconstruction for chest wall resection]. AB - Sixteen patients who involved in chest wall disease underwent major chest wall resection between April 1995 and January 1999. The underlying diseases were 6 recurrence of breast cancers, 4 direct invasion by primary lung cancer, 2 metastatic chest wall tumor, one direct invasion by metastatic lung tumor, one direct invasion by metastatic mediastinal tumor, one radio-induced-necrosis of the chest wall, and one chest wall infection. In 9 patients, the thoracic cage reconstruction was performed using double sheets of absorbable mesh (Dexon mesh), cross string sutures and autologous ribs grafts. None of the patients had major respiratory failure and chest wall unstability. No late complications including infections, pains, recurrence and others related to reconstruction materials have been observed. PMID- 10808291 TI - [Traumatic cardiac tamponade due to the nail stabbing]. AB - A 45-year-old man was admitted to our hospital complaining of mild chest pain. The stab wound on the chest was very small and the nail had been completely buried under the skin. Emergency operation was mandatory for cardiac tamponade by the nail stabbing. Contused right ventricular wall was repaired using Teflon felt under cardiopulmonary bypass. It was important to have a suspicion as cardiac tamponade, when the patient showed uncertain hypotension. PMID- 10808292 TI - [Mistaken grafting to diagonal branch in minimally invasive direct coronary artery bypass: a case required reoperation]. AB - The patient was a 75 year-old male who was admitted with recurrent chest pain during hemodialysis one year after PTCA to the right coronary artery and left circumflex branch (LCX). He had the history of cerebral infarction and chronic renal insufficiency. Coronary angiography showed severe stenosis from the left main trunk to left anterior descending artery (LAD) and restenosis at the PTCA site of LCX. The LCX lesion was dilated with PTCA. Minimally invasive coronary artery bypass (MIDCAB) with left internal thoracic artery (LITA) to LAD was carried out uneventfully. However, chest pain appeared on 1 POD. LITA angiography revealed that LITA was anastomosed to the diagonal branch that had occluded completely in the preoperative angiography. Off-pump CAB to LAD using inferior epigastric artery was carried out through median sternotmy on the same day. He recovered smoothly, and LITA angiography before discharge demonstrated that both grafts to LAD and diagonal branch are patent. PMID- 10808293 TI - [A case report of Binswanger type encephalopathy manifested after coronary artery bypass grafting]. AB - A 68-year-old man showed signs of dementia after undergoing triple CABG with cardiopulmonary bypass. Brain CT revealed widespread atrophy and lucency in white matter without any sign of stroke. No critical stenosis in cervical vessels was found in duplex scanning. He kept politeness well, but had severe memory impairment. These findings were characteristic of Binswanger type encephalopathy which must have been latent preoperatively. This disease is caused by ischemic damage in brain stem and white matter due mainly to atherosclerotic micro cerebrovasculopathy, and highly related to hypertension. We have to be aware of intracranial cerebrovascular disease when assessing the risk of brain damage in candidates for CABG. PMID- 10808294 TI - [Bronchopleural fistula after pneumonectomy following induction therapy for cT4N0M0 squamous cell lung cancer]. AB - A 69-year-old man developed after thoracotomy for lung cancer. He had centrally located cT4N0M0 squamous cell lung cancer that was downstaged to cTxN0M0 by induction chemoradiotherapy with grade 3 hematologic toxicity. Two months after the finish of chemotherapy, right pneumonectomy with concomitant resection of the superior vena cava was performed. Three weeks later, he developed with a bronchopleural fistula and died of ARDS. This demonstrates increased risk of surgery following induction chemoradiotherapy. PMID- 10808295 TI - [Two cases of thymic carcinoma effectively treated by preoperative chemotherapy]. AB - A 67-year-old man was admitted with left pleural effusion on chest X-ray film. Chest CT scans showed an anterior mediastinal tumor, left pleural dissemination and pleural effusion. Percutaneous needle biopsy yielded a diagnosis of epitherial carcinoma. The patient underwent 3 courses of systemic chemotherapy (CDDP + ADM + VCR + CPA). At achievement of partial response, the tumor was completely resected. Examination of the resected tumor revealed partial response to chemotherapy. A 49-year-old woman was admitted our hospital with supraclavicular lymph nodes swelling. Chest CT scans showed an large mediastinal tumor. Percutaneous needle biopsy yielded a diagnosis of squamous cell carcinoma of thymus. The patient underwent 3 courses of systemic chemotherapy. At achievement of partial response, the tumor was completely resected. Examination of the resected tumor revealed partial response to chemotherapy. These cases indicate the usefulness of preoperative chemotherapy for advanced thymic carcinoma to reduce tumor size and to control local invasion, distant metastasis before operation. PMID- 10808296 TI - [Resected case of thymic cyst with primary lung cancer]. AB - Thymic cyst is a rare and benign tumor of mediastinal tumor. A 77-year-old male was pointed out the tumor shadow on Chest X-ray film and diagnosed lung cancer. On admission some examination was performed and revealed mediastinal tumor accidentally. Primary lung cancer was completely resected and mediastinal tumor was extirpated. Pathological examination revealed lung cancer (poorly differentiated squamous cell carcinoma) and thymic cyst. Many methods of examination were tried for diagnosis of thymic cyst, but couldn't diagnose exactly before its resection. Some of the patients with thymic cyst was reported to develop SVC syndrome or change to malignant tumor. Therefore we think complete resection is the best treatment. PMID- 10808297 TI - [Thoracoscopic resection of a pulmonary lymphangioma: a case report]. AB - We report a extremely rare case of a 57-year-old man who was found to have a solitary tumor in the right middle lobe of his lung during an annual check-up. Chest X-ray and CT scan showed a well-circumscribed spherical mass adjacent to the chest wall. The tumor, which was removed by thoracoscopic surgery, was a cystic mass, 17 x 15 mm. Histological examination confirmed the diagnosis of a pulmonary lymphangioma. A brief review of the available literature on this extremely rare type of lung tumor follows the case report. PMID- 10808298 TI - [A case of congenital cystic adenomatoid malformation of the lung in a 4-year-old child]. AB - A case of congenital cystic adenomatoid malformation (CCAM) of the lung is reported. A 3-year-old boy was admitted for elevation of infiltrative shadow in the right lower lung field on a chest X-ray. The clinical diagnosis was emphysematous bullae with inflammation. After 6 months, he had recurrent infected bullae, and was admitted. He received chemotherapy with antibiotics prior to the operation. Right lower lobectomy was performed on February 2, 1998. The resected lung was composed of multiple cysts with thin wall measuring 10-60 mm in diameter. The histological examination of specimen revealed CCAM (Stocker type I), which is rare in this age. The patient has been well for 16 months postoperatively. PMID- 10808299 TI - [Is homocysteine a risk factor for coronary heart disease in patients with terminal renal failure?]. AB - BACKGROUND: Cardiovascular disease is a major cause of mortality in chronic uremic patients. We studied whether homocysteine is an independent cardiovascular risk factor for patients with end-stage renal disease (ESRD). PATIENTS AND METHODS: The study included 163 patients and controls (Group 1: healthy controls, n = 20; Group 2: patients with chronic renal failure, serum creatinine < or = 4 mg/dl, n = 23; Group 3: patients with ESRD, n = 91; Group 4: renal transplant recipients, serum creatinine < or = 2.5 mg/dl, n = 29). We registered patients for the following factors: age, diabetes, smoking, lipids, vitamin B12, folic acid and homocysteine. The coronary heart disease was diagnosed by coronary angiography. RESULTS: The cardiovascular risk profile (hypertension, diabetes, smoking, hyperlipidemia) among uremic patients was significantly increased compared to the healthy controls. There was a significant correlation between the impairment of renal function and the increase of the homocysteine concentration (Group 1: 12 +/- 4.3 mumol/l vs Group 3: 27.8 +/- 15.8 mumol/l; p < 0.001). There was no significant difference of homocysteine between the patients with coronary heart disease and those without (29.9 +/- 18.1 mumol/l vs 26.6 +/- 14.4 mumol/l, not significant). CONCLUSION: In this study a significant correlation between the number of cardiovascular risk factors and the incidence of cardiovascular disease was proven. Although homocysteine was increased among patients with impaired renal function, hyperhomocysteinemia could not be identified as a significant risk factor for coronary heart disease in patients with ESRD. It is assumable that the pathogenesis of coronary heart disease in patients with ESRD is of multifactorial origin. PMID- 10808300 TI - [Asymptomatic bacteriuria]. AB - With the availability of methods to quantitate microbes in urine, one has come to recognize that a certain proportion of apparently healthy individuals have bacteria in their urine in the absence of any clinical symptoms. By definition, asymptomatic bacteriuria is present, if on 1, 2 or more consecutive occasions > 100,000 colony forming units/ml are found in aseptically collected midstream urine, granted that the same microorganism is isolated. Such asymptomatic bacteriuria is found in 2.7% of women aged between 15 and 24 years and increases to 20 to 50% in women above an age of 80 years. In men the prevalence of asymptomatic bacteriuria is considerably lower, but increases to 6 to 20% above the age of 80 years. The microbe most frequently involved in asymptomatic bacteriuria is Escherichia coli, which is characterized by the expression of multiple virulence factors. In this respect they differ from bacteria found in patients with lower or upper urinary tract infections. There is a consensus that in the absence of anatomical or functional abnormalities of the urinary tract asymptomatic bacteriuria per se does not lead to renal scarring, renal dysfunction, or hypertension. The overall prevalence of asymptomatic bacteriuria in pregnancy varies between 2 and 10% with the very small risk of an acute symptomatic episode of urinary tract infection in early pregnancy, but with a substantial risk (30 to 60%) during the last trimester. There is an association between asymptomatic bacteriuria and low birth weight. Asymptomatic bacteriurias are seen in 4 to 60% of immunosuppressed renal graft recipients. The incidence of asymptomatic bacteriuria is also high among patients with diabetes mellitus; the prevalence is approximately 3 fold higher in adult women (7.9 to 11.1%). In these patients asymptomatic bacteriuria does not carry a renal risk. Renal functional prognosis in uncomplicated asymptomatic bacteriuria is excellent. Therefore asymptomatic bacteriuria should be treated during pregnancy, after renal transplantation, prior to urological interventions and in patients with frequent episodes of symptomatic urinary tract infection. PMID- 10808301 TI - [Azathioprine in chronic inflammatory bowel diseases. Evidence base]. AB - AIM: An overview on the evidence-based indications for an immunosuppressive treatment with azathioprine in chronic inflammatory bowel diseases is given. CROHN'S DISEASE: In Crohn's disease, these are the induction of remission in chronic active Crohn's disease, steroid-dependent Crohn's disease, fistulizing Crohn's disease and the maintenance of remission in Crohn's disease. The optimal dose is 2.5 mg/kg body weight, treatment should be maintained for at least 4 years. ULCERATIVE COLITIS: In ulcerative colitis, these are steroid dependency, the maintenance of remission in chronic active ulcerative colitis and the maintenance of remission after induction of remission with cyclosporin or tacrolimus in acute attacks of disease. PMID- 10808302 TI - [Arteriosclerosis and media sclerosis. A comparison of 2 calcifying vascular diseases]. AB - PATHOGENESIS: Arteriosclerosis and Monckeberg's mediasclerosis are vascular diseases associated with calcification of the artery wall. While mediasclerosis in most cases develops in type 2 diabetic patients, arteriosclerosis is the result of a combination of different vascular risk factors. Monckeberg's mediasclerosis typically involves the tunica media, whereas arteriosclerosis associated calcifications primarily involve the intima. CLINICS: Isolated mediasclerosis does not cause narrowing of the blood vessel. The disease is usually asymptomatic, specific therapy has not yet been established. The involvement of the intima in arteriosclerosis finally leads to a decreased circulation. PMID- 10808303 TI - [Diagnosis and therapy of sarcoidosis]. PMID- 10808304 TI - [Acute renal failure in hantavirus infections]. AB - BACKGROUND: The acute renal failure remains a diagnostic challenge for the clinician. CASE REPORTS: Between 1991 and 1996, acute renal failure caused by hantavirus infection was diagnosed in 4 previously healthy male patients. Main symptoms consisted of fever, headache, arthralgia, lumbar and abdominal pain as well as a decline in diuresis. The ultrasonography showed a slight splenomegaly in 2 patients. The clinical chemistry showed elevated serum creatinine from 2.2 mg/dl to 6.7 mg/dl and thrombocytopenia from 4000 to 150,000/microliter. The examination of the urine showed slight proteinuria and microhematuria. The kidney biopsy of 1 patient showed a reversible damage of the tubuli. The pathologic findings normalized within 3 weeks in 3 patients without need for dialysis. One patient developed a severe clinical course with acute renal failure and pulmonary edema requiring dialysis. In all patients, the renal function improved. PMID- 10808305 TI - [Recurrent pneumothorax in interstitial lung disease, anogenital and abdominal wall exanthema and secondary amenorrhea in a 20-year-old patient. Langerhans-cell histiocytosis]. AB - CASE REPORT: A 20-year-old female patient was admitted to the hospital because of a pneumothorax. A secondary amenorrhea was coexisting. On the abdominal wall and in the anogenital area a maculopapular exanthema was visible. Chest X-ray and HR CT showed an interstitial lung disease with high probability of histiocytosis X which was confirmed by biopsies of lung and skin tissue. The cause of amenorrhea was a histiocytic infiltration of the pituitary stalk. Conventional "consensus treatment" for adults with vinblastine, prednisone and etoposide did not prevent relapsing bilateral pneumothoraces. Treatment was successful with the protocol as proposed by the international study for children. This treatment is characterized by higher dosage and combination with 6-mercaptopurine. PMID- 10808306 TI - [Primary pigmented nodular adrenocortical dysplasia (PPNAD) within the scope of Carney complex as the etiology of Cushing syndrome]. AB - BACKGROUND: Primary pigmented adrenocortical dysplasia (PPNAD) represents a rare disorder of the adrenal glands and frequently occurs in patients with the so called Carney complex. Carney complex is an autosomal dominant neoplasia syndrome including skin and mucosal lentigines, myxomas, and PPNAD. CASE REPORT: A 37-year old woman suffered from several episodes of weight gain/depression and weight loss/mania indicating cyclic hypercortisolism. Finally, she developed a full blown Cushing's syndrome (CS) treated by bilateral adrenalectomy. She had PPNAD in the setting of Carney complex. CONCLUSION: PPNAD may lead to different clinical manifestations: 1. subclinical hypercortisolism, 2. intermittent hypercortisolemia, and 3. full-blown CS. It can be diagnosed with the 6-day Liddle test that typically shows a paradoxical stimulation of cortisol secretion after dexamethasone administration. The treatment of choice for PPNAD is bilateral adrenalectomy in order to prevent devastating long-term effects of hypercortisolism. PMID- 10808307 TI - [Recurrent "cold abscesses" with spontaneous perforation as a rare course in abdominal tuberculosis]. PMID- 10808308 TI - [Habilitation in medicine--only pro forma?]. PMID- 10808309 TI - [Medical habilitation. Accepted academic qualifications or outdated formalism?]. AB - BACKGROUND: With the changes in the structure of education and society, in recent years also the medical habilitation (German postdoctoral lecturing qualification) has been called into question as an academic qualification. With this in mind, the aim of our study was to discover the current opinion of those who had successfully completed a habilitation on the prerequisites for a habilitation, the habilitation procedure and the status of a habilitation, and to document potential wishes for reform. COLLECTIVE AND METHODS: The target group of our survey were the 616 persons (female: 77, male 539) who successfully completed their habilitation in 1997 at one of the 36 German medical faculties. The database was formed by an anonymous questionnaire (23 items), which included questions on sociodemographic factors and occupational history (general part), and subjective opinions (specific part). Recruitment of the participants in the survey and passing on of the questionnaire were carried out by the office of the medical dean of the various universities, as the names of those who had completed a habilitation were not available to the investigators for reasons of protecting the individual's rights and identity. Evaluation of the returned questionnaires was carried out using descriptive statistics. Subgroups were formed according to sex, age and subject groups. RESULTS: The questionnaire was answered in a useable form by 389 persons (female: 46, male 343) from 35 medical faculties (return quota 63%). 95% of those who took part in the survey were registered doctors; 79% of these came from clinics, 16% from the field of theoretical medicine. 81% were specialist physicians. 5% had studied the natural sciences or humanities. The median age at the time of completing the habilitation was 38 years (minimum 30, maximum 54 years old). At present 93% were assistant professors, 5% were professors. The median interval between the doctoral thesis and habilitation was 10 years. 58% had carried out a period of research abroad. In 90% of cases the persons had written a postdoctoral thesis for their habilitation, 10% qualified cumulatively. 47% had improved their occupational rank within a period of 2 years after completing the habilitation, about 2/3 of these reached senior positions. Among the prerequisites for habilitation, "Humboldt's trias" (research, teaching and caring for patients) was accepted by the great majority. Other prerequisites regarded as important for habilitation were publications, holding talks, specialist status and experience abroad. Impact factors, however, should be regarded as important conditions for habilitation only in combination with other criteria. The value of a habilitation was not called into question; 89% would recommend completing one. 80% of those questioned, however, thought the procedure for completing a habilitation should be optimized. The general abolition of the medical habilitation was, however, not desired by the vast majority. CONCLUSION: The value of completing a medical habilitation is not a point for debate for most of those who successfully completed one. It remains the springboard for occupational advancement. The vast majority do not wish to see it abolished. Also the usual prerequisites for habilitation are accepted by the majority of persons. The procedure for completing a habilitation is, however, regarded as in need of improvement. There is a wide consensus of opinion regarding potential aims for reform. PMID- 10808310 TI - [CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy): a new genetic disease of the cerebral arteries associated with vascular leukoencephalopathies]. PMID- 10808311 TI - [Vaccinal prophylaxis in the 19th century]. PMID- 10808312 TI - [Hypereosinophilia]. PMID- 10808313 TI - [Structure and function of polynuclear eosinophils]. AB - Eosinophils are mainly tissue cells, attracted at mucosal sites by several chemotactic factors. They express numerous surface receptors, which make them sensitive to several differentiation, activation and inflammatory signals. Eosinophilic specific granules contain highly cationic proteins, which are released upon activation by immune effectors, like immune complexes or by inflammatory mediators. These basic granule proteins are cytotoxic for non-self targets such as parasite larvae but also for normal cells or tissues, giving to eosinophils a dual function in immune defense and in immunopathology. Eosinophils are also a source of various mediators including growth factors, proinflammatory cytokines, chemokines, and immunoregulatory molecules, conferring to eosinophils multiple functions, not only as effector and pro-inflammatory cells but also as immunoregulatory cells. PMID- 10808314 TI - [Reactive hypereosinophilia in parasitic diseases]. AB - Hypereosinophilia is often linked to the presence of pluricellular parasites in host tissues. Polynuclear eosinophils are sprung from the bone marrow. After multiplying and maturing, they are thrown into the blood flow and from the blood to the tissues where they are found in immediate contact with the parasite. Eosinophils are major components of the parasitic granuloma. Parasitic diseases are a major cause of hypereosinophilia and eosinophilia is mainly due to helminths. Protozoa do not produce hypereosinophilia, except for toxoplasmosis in which a low and discontinuous eosinophilia may be seen. Subsequently, maggots producing myiasis yield to hypereosinophilia too. In helminthiasis, the action of the eosinophil granulocyte is double. In tissues, it destroys the parasite and plays a regulatory role in mastocytes degranulation. Eosinophils which participate in the inflammatory reaction secrete factors which neutralize mediators liberated by mastocytes, histamine mainly, destroyed by histaminase. In a practical point of view, blood hypereosinophilia is a very useful tool for diagnosis. Eosinophilia reach early a high value, this before the parasitic infection becomes detectable by means of resources other than immunological. The eosinophils rates decreases rapidly as an effect of the anthelmintic drug, this confirming the efficacy and specificity of the prescribed treatment. PMID- 10808315 TI - [Hypereosinophilic reactions in hypersensitivity states]. AB - Among eosinophil-related diseases, many of them are linked to hypersensitivity mechanisms which are dependent or independent on the development of an IgE response. In certain patients a clear relationship can be identified with an environmental airborne allergen: allergic disorders associated with the presence of eosinophils present in blood or in the airway lumen include allergic asthma, perennial or seasonal allergic rhinitis, less often allergic sinusitis or some cases of chronic cough. In other clinical situations, proofs for a direct involvement of IgE are difficult to establish, for example in drug-induced pulmonary eosinophilias. In allergic bronchopulmonary aspergillosis, two distinct mechanisms are implicated: an IgE dependent process but also the proliferation of the fungal microorganism into the bronchial tree. Similarly in Churg++ and Strauss syndrome, it does not exist any demonstration for an IgE mechanism, although a large majority of patients are atopic, with high serum total IgE levels. So it appears that the border between IgE and non IgE related disorders remains at the present time poorly defined. PMID- 10808316 TI - [Hypereosinophilic reactions in cancer]. AB - Hypereosinophilia may be seen in the blood or tissues of cancer patients. A part of lymphomas, the tumors more frequently involved in such eosinophilia, are digestive and respiratory tract carcinomas. The prevalence of cancer-associated eosinophilia is very variable among the different kind of tumors. These cancer associated eosinophilia are the results of the secretion by tumor or inflammatory cells of interleukin-3 and -5 and GM-CSF. The anti-cancer properties of eosinophilic polymorphonuclear are poorly understood and the prognostic significance of cancer-associated blood or tissue eosinophilia has not been assessed by controlled studies. PMID- 10808317 TI - [Idiopathic hypereosinophilic syndrome]. AB - The idiopathic hypereosinophilic syndrome is defined by the combination of prolonged eosinophilia and evidence of tissue damage including heart and nervous system. Cardiac disease is the major cause of mortality. The idiopathic hypereosinophilic syndrome is a heterogeneous collection of disorders. Products of activated eosinophils could exert toxicities on specific organs; it is a diagnosis of exclusion. There is no specific tests diagnostics. The development of a comprehensive diagnosis and therapeutic approach ensure from a better understanding of the pathogenesis. PMID- 10808318 TI - [Chronic hypereosinophilia: a model of Th2 disorders]. AB - Interleukin-5 produced by Th2-type lymphocytes is involved in the pathogenesis of a number of hypereosinophilic disorders. We and others have identified clonal Th2 cells with abnormal surface phenotypes in peripheral blood of certain patients presenting the idiopathic hypereosinophilic syndrome. We took advantage of the CD3- CD4+ phenotype of our patients' T cells to determine the activation signals involved in their production of Th2 cytokines and expansion, independently of T cell receptor engagement. In vitro cocultures performed with dendritic cells demonstrated the critical role of co-stimulatory signalling through B.7/CD28 and LFA-3/CD2 pathways and the involvement of an autocrine IL2/IL2R loop in the activation of these Th2-type cells. The high-level spontaneous apoptosis displayed by these cells in vitro was drastically inhibited by IL2 and IFN-alpha. New therapeutic strategies could result from our observations. Indeed, the hypereosinophilic syndrome may represent an unexpected application of new immunomodulatory molecules such as CTLA4-Ig and anti-IL2R-alpha. PMID- 10808319 TI - [Chronic hypereosinophilia and hematologic malignancies]. AB - Peripheral blood and/(or) marrow hypereosinophilia may be associated to various subtypes of myeloid or lymphoid hematological malignancies. Eosinophilia is frequently paraclonal, due to the recruitment of eosinophilic polymorphonuclear cells by cytokines secreted by lymphomatous or leukemic cells. In some myeloid disorders, eosinophilia may also be clonal, due to the involvement of the eosinophilic lineage within the leukemic clone. Generally, hypereosinophilia is only a side-symptom and eosinophilic visceral damages are rare. PMID- 10808320 TI - [Tissue hypereosinophilia]. AB - Eosinophils are produced in the bone marrow and circulate in the peripheral blood in a quiescent state. After margination and emigration into tissues through postcapillary venules, eosinophils primarily reside in mucosal sites including the gastrointestinal and respiratory tracts. Eosinophilic gastroenteritis and the idiopathic chronic eosinophilic pneumonia, first described by Carrington, are the most characterized idiopathic diseases defined by an eosinophil infiltration restricted respectively to the gastrointestinal and the respiratory tracts. Differential diagnosis can be difficult since several disorders and particularly the idiopathic hypereosinophilic syndrome have many clinical, morphological and histological similarities. PMID- 10808321 TI - [The United Kingdom, New Zealand ... and us]. PMID- 10808322 TI - [Nosocomial infections. Epidemiology, diagnostic criteria, prevention, treatment principles]. PMID- 10808323 TI - [Tumors of the ovarian epithelial lining. Pathologic anatomy, diagnosis, development, principles of treatment]. PMID- 10808324 TI - [Infections with dermatophytes of the hairless skin and folds. Diagnosis and treatment]. PMID- 10808325 TI - [Carpal tunnel syndrome. Etiology, diagnosis]. PMID- 10808326 TI - [Hallucinations. Diagnostic orientation]. PMID- 10808327 TI - [Palliative and supportive care. Work setting and ethical aspects]. PMID- 10808328 TI - [Kidney cancer in the adult. Pathologic anatomy, diagnosis, development, principles of treatment]. PMID- 10808329 TI - [Disseminated intravascular coagulation. Etiology and diagnosis]. PMID- 10808330 TI - The secretory pathway from history to the state of the art. AB - Maintenance of the structural and functional organization of a eucaryotic cell requires the correct targeting of proteins and lipids to their destinations. This is achieved by the delivery of newly synthesized material along the secretory pathway on one hand and by the retrieval of membranes on the other hand. Various models have been suggested over the years to explain traffic flow within the secretory pathway. The only two models that are under discussion to date are the "vesicular model" and the "cisternal maturation model". A wealth of information from various experimental approaches, strongly supports the vesicular model as the general mode of intracellular transport. Three major types of protein-coated transport vesicles are characterized in molecular detail, and have been attributed to various steps of the secretory pathway: COPII-coated vesicles allow exit from the endoplasmic reticulum (ER), COPI-coated vesicles carry proteins within the early secretory pathway, i.e. between ER and Golgi apparatus, and clathrin-coated vesicles mediate transport from the trans-Golgi network (TGN). In this review we will give an overview of the route of a protein along the secretory pathway and summarize the progress that was made within the last decades in the characterization of distinct intracellular transport steps. We will discuss the current models for the formation and fusion of vesicular carriers with a major focus on the mechanism underlying budding of a COPI-coated vesicle. PMID- 10808331 TI - Neurotoxins as tools in dissecting the exocytic machinery. PMID- 10808332 TI - Annexins and membrane fusion. PMID- 10808333 TI - The full complement of yeast Ypt/Rab-GTPases and their involvement in exo- and endocytic trafficking. PMID- 10808334 TI - Possible roles of long-chain fatty Acyl-CoA esters in the fusion of biomembranes. PMID- 10808335 TI - Brefeldin A revealing the fundamental principles governing membrane dynamics and protein transport. PMID- 10808336 TI - Membrane fusion events during nuclear envelope assembly. PMID- 10808337 TI - Transactions at the peroxisomal membrane. PMID- 10808338 TI - Neurons, chromaffin cells and membrane fusion. PMID- 10808339 TI - Reversibility in fusion protein conformational changes. The intriguing case of rhabdovirus-induced membrane fusion. PMID- 10808340 TI - Specific roles for lipids in virus fusion and exit. Examples from the alphaviruses. PMID- 10808341 TI - Fusion mediated by the HIV-1 envelope protein. PMID- 10808342 TI - Sulfhydryl involvement in fusion mechanisms. PMID- 10808343 TI - Using mind-body techniques in dentistry. AB - Mind-body medicine is a win-win proposition for dentists and their patients. Patients experience less anxiety and less discomfort during procedures. However, it's important to know the advantages and disadvantages if you are considering offering mind-body services in your practice. PMID- 10808344 TI - Crown lengthening to facilitate restorative treatment in the presence of incomplete passive eruption. AB - The following clinical case involves subgingival caries noted in the presence of incomplete passive eruption in the maxillary anterior sextant. Understand the clinical entity known as incomplete passive eruption and discover utilization of crown lengthening surgery to treat this condition. PMID- 10808345 TI - Microdentistry: a new standard of care. AB - Micro-dentistry offers a new standard of care to address and solve the current challenges facing the dental profession. Patients will be grateful for accurate diagnosis of the pit and fissure system with caries detection dye and magnification, followed with comfortable air abrasion debridement and preparation for restoration with an adhesive composite material. PMID- 10808346 TI - Latex allergy: a potential liability issue. PMID- 10808347 TI - Periodontal treatment for elderly patients. AB - The largest growing segment of our society is that of older Americans. The population explosion of older adults challenges both general practitioners and periodontists to provide the highest level of care available. More of our patients will have concurrent medical conditions that alter or modify the delivery and provision of periodontal care. This paper reviews some of the common conditions occurring in the older patient and suggests some modifications in periodontal therapy which may be beneficial. The therapist must be knowledgeable about the medications commonly used for treating the chronic diseases of the older population. Finally, the therapist must become adept in performing functional assessments of patients so that the types of therapy administered contribute to the quality of life desired by patients. PMID- 10808348 TI - Practical considerations in intraoral digital radiography. PMID- 10808349 TI - The cox-2 inhibitors. PMID- 10808350 TI - Management of occupational exposures to bloodborne viruses: HIV post-exposure prophylaxis. AB - Unlike exposure to hepatitis B-infected blood, where risk of transmission is low and post-exposure prophylaxis is safe and effective, exposure to HIV is much different. This risk of HIV transmission is extremely low and the safety and efficacy of chemoprophylaxis is uncertain. Dental health care workers should be made aware that the potential side effects and toxicity of the standard post exposure drug regimen outweigh the negligible risk of transmission posed by the exposure. PMID- 10808351 TI - Infected cyst secondary to mandibular bone plate and screw. PMID- 10808352 TI - Endodontics emergency treatment. PMID- 10808353 TI - Extra-oral approach for the removal of an impacted mandibular canine. PMID- 10808354 TI - Spontaneous disappearance of an eruption cyst. PMID- 10808355 TI - A study on how military dependents at Philippine Navy Officers Wives Association (PNOWA) Child Learning Center brush their teeth. 2. PMID- 10808356 TI - Sugar content of commonly eaten snack foods of school children in relation to their dental health status. AB - Sugar is the principal substrate for micro-organisms causing dental caries. Yet it is impossible to eliminate sugar in the diet as almost all available foods particularly snack foods contain sugar. WHO recommended 10 kg. sugar or 27.40 gms. per person per year in non-fluoridated area and 15 kg. sugar/person year in non-flouridated areas for low caries prevalence. The FFQ and 24-Hour Food recall of 824 rural and urban schoolchildren showed that all their snacks foods are high in sugar except gelatine. Their sugar intake was three times more than the WHO recommendation with a mean daily intake of 90 gms. for urban and 88 grams of sugar for the rural. White sugar, hard candies, raisins, coco jam, and milk choco were the five most commonly eaten snack foods with high sugar content. Oral examination of the schoolchildren showed that dental caries was higher in the urban than in the rural. As a whole, the urban schoolchildren consumed more sugar and had higher dental caries compared to the rural schoolchildren. No significant correlation was found between sugar intake and dental caries. PMID- 10808357 TI - Impactions in orthodontics. PMID- 10808358 TI - Dental caries and periodontal disease (prevention and control methods). AB - There is a compelling need to apply preventive programs in both private and community practice of dentistry. This is to maintain improvements in oral health in developed and industrialized countries, and to stem increases in oral diseases in underserved and developing ones. At the outset, the terms prevention and control must be understood. The former is considered to mean a procedure or course of action that prevents the onset of disease, whereas the latter, implies reversing or stabilizing disease conditions. To be more precise, prevention will refer to the pre-pathologic or pre-clinical stage encompassing the promotive and specific protection levels--primary prevention stage. On the other hand, control will encompass early diagnosis and prompt treatment, disability limitation and rehabilitation levels-termed also collectively, as pathologic, clinical and final stages, or secondary and tertiary prevention. Community-based programs are usually structured to compliment therapeutic interventions of oral diseases, as well as prevention. In this era, and towards the next millennium, preventive and control programs are given high priorities in order to minimize the need for curative, restorative and therapeutic management of oral diseases. This review of the literature will give emphasis on established methods and programs for the prevention and control of the two most common oral diseases, dental caries and periodontal disease. The problems, background, and oral health objectives for the year 2000 as proposed by the World Health Organization (WHO) and the Federation Dentaire Internationale (FDI), as well as the recent advances in oral health relative to these diseases will be discussed. Finally, to better improve the efficacy of existing prevention and control methods, research needs and areas of concern relative to these diseases will be given consideration. PMID- 10808359 TI - Pleomorphic adenoma of the submandibular gland. PMID- 10808360 TI - Management of cutaneous fistula secondary to third molar infection. PMID- 10808362 TI - Results of the recent TDA member survey. AB - This article fully outlines active TDA members' responses to the various questions posed by the recent membership survey. Every effort was made to ensure the accuracy of the results. To this end, a professional research firm was employed to cross tabulate answers and analyze the data. A thorough reading will provide you with the TDA members' perspective on a wide range of important dental issues. PMID- 10808361 TI - The supply of dentists in the Philippines, a situationer. AB - This study is a review of secondary data on the supply of dentists in the Philippines from information gathered from various governmental and non governmental agencies. It was done to determine general trends in the supply and stock of dentists as well as have an overview of the oral health human resources in the country. Initial findings indicate an unequal distribution of dentists as well as a large amount of wastage in the production of dentists. PMID- 10808363 TI - Clinicopathologic correlation quiz: solitary elevated lesions of the lower lip. PMID- 10808364 TI - Interleukin-1 family expression in human breast cancer: interleukin-1 receptor antagonist. AB - We hypothesize that interleukin-1 alpha, beta, and receptor antagonist (IL-1 alpha, IL-1 beta, and IL-1 ra, respectively) are present and tumor cell associated in human breast cancer (HBC). We believe the levels of these cytokines in breast tumor homogenates relate to other known prognosticators of patient survival (i.e., estrogen receptor [ER] status). Our results demonstrated that, immunohistochemically, tumor cells express IL-1 alpha, IL-1 beta, and IL-1 ra in most specimens tested. In breast tissue homogenates, IL-1 alpha levels correlated inversely with ER levels (p < 0.06), whereas IL-1 ra levels correlated directly with both ER levels (p < 0.009) and IL-1 beta levels (p < 0.06). When analyzing cytokine levels for the ER (-) versus ER (+) patient groups, we found that in many instances these groups showed a different cytokine profile. These studies suggest that the IL-1 family of cytokines may be important in regulating protumorigenic activities within the HBC tumor microenvironment. PMID- 10808365 TI - Non-Hodgkin's lymphoma: an analysis of the Metropolitan Detroit SEER database. AB - We evaluated incidence and survival trends of non-Hodgkin's lymphoma (NHL) in a large population-based cancer registry. Data regarding demographics, histology, incidence, and survival were obtained on all patients with NHL registered in the Metropolitan Detroit Cancer Surveillance System, a participant in the Surveillance Epidemiology and End Results (SEER) Program of the National Cancer Institute. Incidence and survival trends from 1973 through 1995 were evaluated and stratified based on age at diagnosis, sex, race, and tumor grade. There were 11,978 patients diagnosed with NHL and recorded in the Metropolitan Detroit SEER registry from 1973 to 1995. The age-adjusted incidence rate increased from 8.6 to 15.8 per 100,000, leading to an overall increase in incidence of 83% and an average annual increase of 3.2% per year. Incidence increased significantly (p < 0.05) over time in all age groups except the youngest (ages 0-19) and in all demographic groups studied. Incidence was highest in white men and lowest in black women. The incidence of both low-grade and intermediate/high-grade NHL increased significantly for each age group (p < 0.05) except the youngest (ages 0 19). In the oldest patients (70+ years), the incidence of intermediate/high-grade NHL was almost double that of low-grade NHL. Five-year relative survival increased from 64% (1973-1983) to 68% (1984-1991) for patients with low-grade NHL and from 40% to 44% for those with intermediate/high-grade NHL. The increase in relative survival was only seen in whites, however, with 5-year relative survival in blacks decreased from 53% (1973-1983) to 45% (1984-1991). In metropolitan Detroit, the current NHL epidemic affects all age groups except the very young (ages 0-19), both sexes, and both whites and blacks and is due to increases in the incidence of both low-grade and intermediate/high-grade NHL. Five-year survival rates have increased for whites but not for blacks. PMID- 10808366 TI - Unresectable adenocarcinoma of the pancreas: patterns of failure and treatment results. AB - We retrospectively reviewed the outcome of 36 consecutive patients seen in our department with nonmetastatic unresectable adenocarcinoma of the pancreas. There were 28 men and 8 women with a mean age of 63.7 years (range, 43-78 years). Radiation therapy (RT) was given to all patients. 5-Fluorouracil (5-FU) was given as a bolus (500-1000 mg/m2/day) during the first and last weeks of RT in 22 patients, whereas continuous 5-FU (225 mg/m2/day) was given to 3 patients. The median survival for all patients was 10 months. Overall survival for all patients at 1 and 3 years was 47.2% and 19.1%, respectively. The pancreas, peritoneum, liver, and lungs were components of failure in 66.7%, 33.3%, 14.8%, and 14.8%, respectively. Local control was a major problem in patients treated for unresectable pancreatic cancer. The results of this study confirm the dismal prognosis of these patients. PMID- 10808367 TI - Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography: a time and concentration study. AB - Platinum-based chemotherapies cause the formation of DNA adducts and have profound effects on DNA. This study measured cis-diamminedichloroplatinum II (cisplatin) DNA adducts by 32P-radiolabeling DNA, enzymatically digesting radiolabeled DNA, separating the formed adducts on two-dimensional thin-layer chromatography, and quantitating the adducts with autoradiography and densitometry. HeLa DNA was incubated with cisplatin at varying concentrations (6.25-325 nM) and times (0 min to 72 hr). Cisplatin rapidly depurinated dGMP and dAMP (90%, 15-min incubation with 325 nM cisplatin). Partial depurination of dGMP (15%) and dAMP (25%) occurred with lower cisplatin concentrations at equal incubation times. A minimum of four new adducts, with relatively rapid migratory patterns, were detected at high cisplatin concentrations with short incubation times. These results indicate that the depurination of DNA correlates with DNA adduct formation and that the quantification of these adducts may be applicable to monitoring tumor and host cell response to cisplatin chemotherapy. PMID- 10808368 TI - The antineoplastic agent paclitaxel (Taxol) increases contractile activity in human saphenous veins and human mammary arteries. AB - Effects of the antineoplastic agent paclitaxel (Taxol) were studied on contractions of isolated human saphenous vein (HSV) and mammary artery (HMA). Peak force developed by vascular segments with cumulative concentrations of physiologic agonists was enhanced by paclitaxel, producing a shift to the left of dose-response curves. Paclitaxel 1 microM decreased ED50 (in microM) for norepinephrine from 1.01 +/- 0.24 to 0.20 +/- 0.06 (n = 16, p < 0.05) in HSV and from 1.30 +/- 0.30 to 0.51 +/- 0.21 (n = 15, p < 0.05) in HMA and for 5 hydroxytriptamine from 0.64 +/- 0.19 to 0.21 +/- 0.07 (n = 20, p < 0.05) in HSV. Paclitaxel 1 microM also significantly increased the peak force of contractions elicited by endothelin-1 0.01 microM in HSV. In contrast, it did not affect contractions evoked by KCl 80 mM. These results show that paclitaxel produces a hyperreactivity in human vessels challenged by physiologic agonists, which suggests that administration of paclitaxel to patients could augment peripheral resistance and increase blood pressure. PMID- 10808369 TI - p185HER2 overexpression in human breast cancer using molecular and immunohistochemical methods. AB - With the successful clinical trials of the engineered antibody Herceptin (in advanced-stage breast cancer) and adriamycin-based chemotherapy regimens (in the adjuvant setting), the need to detect p185HER2 overexpression or associated amplification of the coding gene HER2 in breast cancer patients is escalating. Twenty to 30% of breast carcinomas have overexpression of p185HER2. This condition correlates with poor patient prognosis and predicts response to chemotherapy in lymph node-positive patients. In this study we compare quantitation of p185HER2 in breast cancer at the gene and protein levels using differential polymerase chain reaction (PCR) and immunohistochemistry, respectively. To assign HER2 gene copy numbers, a calibration curve was constructed using normal breast epithelia and breast carcinoma cell lines having known dosages of amplified HER2. We found corresponding molecular and immunohistochemical results in 85% of the 13 paraffin-embedded breast carcinoma cases examined. Two cases were found to have minimum gene amplification but marked p185HER2 overexpression, suggesting an alternative mechanism to overexpression such as transcriptional activation. Although the differential PCR assay exhibits saturation approaching 20 HER2 gene copies, this may not be clinically significant because the immunohistochemical assay also appears to saturate in this gene copy number range. PMID- 10808370 TI - Postradiation squamous cell cancer of the breast. AB - Primary squamous cell carcinoma of the breast is a rare clinical entity. We diagnosed a patient who presented with a pure squamous cell cancer of the breast 12 years after local radiation for a primary adenocarcinoma of breast. Previously reported cancers related to radiation of the breast have been squamous cell of skin and esophagus and sarcomas, especially angiosarcomas, and on literature review this is the first reported case of primary squamous carcinoma of the breast related to previous radiation. Although our observation suggests a possible link, no established cause and effect relationship is known at this point. PMID- 10808371 TI - Histamine: a novel approach to cancer immunotherapy. AB - The functions of intratumoral lymphocytes in many human malignant tumors are inhibited by reactive oxygen species (ROS), generated by adjacent monocytes/macrophages (MO). In vitro data suggest that immunotherapeutic cytokines such as interleukin-2 (IL-2) or interferon-alpha (IFN-alpha) only weakly activate T cells or natural killer (NK) cells in a reconstituted environment of oxidative stress and that inhibitors of the formation of ROS or scavengers of ROS synergize with IL-2 and IFN-alpha to activate T cells and NK cells. In this review, we focus on the immunoenhancing properties of histamine, a biogenic amine. Histamine inhibits ROS formation in MO via H2-receptors; thereby, histamine protects NK cells from MO-mediated inhibition and synergizes with IL-2 and IFN-alpha to induce killing of NK cell-sensitive human tumor cells in vitro. Histamine also optimizes cytokine-induced activation of several subsets of T cells by affording protection against MO-inflicted oxidative inhibition. The putative clinical benefit of histamine as an adjunct to immunotherapy with IL-2 and/or IFN-alpha is currently evaluated in clinical trials in metastatic malignant melanoma and acute myelogenous leukemia. PMID- 10808372 TI - BCL-6 in the pathogenesis of non-Hodgkin's lymphoma. PMID- 10808373 TI - Use of hair dyes, hematopoietic neoplasms, and lymphomas: a literature review. I. Leukemias and myelodysplastic syndromes. AB - We review published epidemiologic studies on personal use of hair dyes and leukemias and myelodysplastic syndromes (MDS). A subsequent article will review studies on lymphomas and multiple myeloma. A computerized literature search for the years 1966 through 1996 was completed using MEDLINE. Data were extracted using a standardized form that recorded study design, study population, type of cases, comparison group, sources of data on personal exposure to hair dyes, method of data collection, type of exposure data collected, covariates, and results. The above search identified 13 epidemiologic studies on the possible association between personal use of hair dyes and leukemias and MDS. Although there are some reports of positive associations, overall the evidence linking personal use of hair dyes to various leukemia and MDS subgroups is weak. One cannot definitively rule out an association, however, because of the methodologic limitations, such as small numbers of exposed cases and lack of detailed exposure information. Any further research would need much better assessment of hair dye use, including product type, color frequency, duration, and changes in use over time, and adequate statistical power. PMID- 10808374 TI - Diagnostic evaluation of neurologic metastases. PMID- 10808375 TI - Introduction: management of cancer in the Canadian Healthcare System. PMID- 10808376 TI - Cancer care in Ontario: accessible oncology on sale, or cheque's in the mail? PMID- 10808377 TI - Canadian healthcare system's management of patients with cancer. PMID- 10808378 TI - Unresectable adenocarcinoma of the pancreas. PMID- 10808380 TI - Society of General Internal Medicine 23rd annual meeting. Boston, Massachusetts, USA. May 4-6, 2000. Abstracts. PMID- 10808381 TI - BOSTON'99, the first joint meeting of the Behavioral Pharmacology Society (43rd annual meeting) and the European Behavioral Pharmacology Society (8th biennial meeting). Boston, Massachusetts, USA. 1-4 September 1999. Abstracts. PMID- 10808379 TI - Hair dyes, cancer, and epidemiology. PMID- 10808382 TI - [Gene therapy of neoplasms]. PMID- 10808383 TI - [Drug compliance in patients treated with antibiotics in an ambulatory setting for respiratory tract infections]. AB - Patient non-compliance is a phenomenon which leads to less effective and more expensive therapy, particularly in general practice conditions. One of the most common problem of out-patients care are respiratory infections. The aim of this study was to evaluate the extent of patient non-compliance among out-patients treated with short-time antibiotic for respiratory tract infections and to find factors associated with patient adherence. Two methods of compliance measurement were used: the questionnaire method and pill count. The study group consisted of 154 out-patients with respiratory infections. The pill count method revealed that 46.8% of patients did not take their antibiotics as prescribed, and only 9.7% stated that in inquiry. Patient compliance was associated with such factors as being married, subjective estimation of health state as lower then peers, present chronic diseases, present ischemic heart disease, subjective estimation of disease as "serious", localisation of infection in lower respiratory tract, dosage schedule, the number of drugs taken at examination > 2, the number of drugs doses taken at examination > 6 and the number of drugs taken longer then a week > 2. Age, gender, education and professional activity did not influence the level of compliance. The study points out usefulness of both inquiry and pill count method in revealing patient non-compliance in conditions of general practice. PMID- 10808384 TI - [The effect of low molecular weight heparin on peripheral leukocyte chemotaxis in patients with asthma]. AB - Evidence has now accumulated that heparin can significantly affect immune response including allergic inflammation. Cell motility is supposed to be very crucial in this process. Thus the aim of that study was to investigate whether heparin is chemoattractant for some inflammatory cells in asthmatics. Peripheral blood mononuclear cells and neutrophils from 6 healthy subjects and 16 asthmatics were obtained by gradient centrifugation. Chemotaxis assays towards low molecular weight heparin-Clexane were performed in Boyden chambers. We found that heparin is chemoattractant for both PBMCs and neutrophils in the wide concentration range (1-2000 micrograms/ml)--maximal chemotaxis was observed at concentrations 50-500 micrograms/ml. Chemotactic motility of cells from asthmatics was weaker than from healthy subjects; the difference wasn't significant. PMID- 10808385 TI - [Early, targeted population based screening for COPD. Preliminary study]. AB - COPD is the fourth leading cause of death in Poland, unfortunately diagnosed not early enough. The aim of the study was to establish prevalence of COPD in chronic smokers. Therefore, using daily press and TV, smokers with at least 10 year history of smoking, over 40 years of age, were invited for a free spirometry. 263 subjects (177 M and 86 F) mean age 54 +/- 0.6 years were examined. Most of them (97.7%) were smokers with a history of 32.2 +/- 0.9 pack-years, 6 persons (2.3%) were passive smokers. 110 persons (41.8%) presented bronchial obstruction, the remaining (58.2%) had normal spirometric values. Following recommendations of the Polish Society of Physio-pneumonology bronchial obstruction was classified as mild in 25.1%,- moderate in 12.1% and severe in 4.6% subjects. Majority of examined subjects presented with COPD symptoms, cough (62.7%), expectoration (68.8%) and dyspnoea (50.2%). The presence of those symptoms did not differ among groups with different severity of bronchial obstruction. However, there were significant differences in age (p < 0.05) and years of smoking habit (p < 0.01). The great efficacy of targeted screening for COPD (40%) should be an incentive to perform routine spirometric examination in smokers with more than 20 years of smoking history. PMID- 10808386 TI - [BAL from two different lung segments indicated by high resolution computed tomography (HRCT) in patients with sarcoidosis. I. Evaluation of alveolitis homogeneity and estimation of HRCT usefulness in selection of lung region for BAL]. AB - The aim of the study was: 1) to evaluate the homogeneity of alveolitis by estimation of lymphocytes subsets in double BAL (2 x 120 ml) from two different lung segments: with the most (s.A) and with the least (s.B.) extensive involvement estimated by high resolution computed tomography (HRCT) 2) to examine the usefulness of HRCT as a guide method in selection the most reliable lung region for BAL. Examined group consisted of 28 sarcoid patients with homogeneous, regular distribution of nodular opacities in conventional chest X-ray (14 F, 14 M aged 19-54). Twelve patients showed homogeneous distribution of HRCT changes (RD) in lung parenchyma and 16 showed nonhomogeneous distribution of HRCT changes (ND) with domination of pathological changes in upper lobes. Eleven healthy volunteers served as controls. BAL lymphocytes subpopulations (CD3, CD19, NK, CD4, CD8, HLA DR, CD25,) were estimated by flow-cytometry. Among patients from ND group in BAL from s.A we found the significantly higher (p < 0.01) mean total cell yield, the significantly higher (p < 0.05) mean values of % of lymphocytes (45.2 vs 36.8%) and CD4/CD8 ratio (5.3 vs 4.4) than in BAL from s.B. Also the mean values of absolute number of lymphocytes and lymphocytes CD4, HLA-DR, CD25 in ND group were significantly higher in BAL from s.A than in BAL from s.B. In RD group and in controls no significant differences between BAL findings from s.A and s.B were noticed. Our results suggest that: 1) alveolitis process is not fully homogeneous, it's intensity is greater in upper lobes with most extensive involvement of HRCT changes 2) HRCT can serve as a useful method for selection the most reliable lung region for BAL. PMID- 10808387 TI - [BAL from two different lung segments indicated by high resolution computed tomography (HRCT) in patients with sarcoidosis. II. The role of T gamma delta lymphocytes (T gamma delta)]. AB - The aim of the study was to evaluate the role of lymphocytes T gd in sarcoidosis by estimation of T gd cells in double BAL (2 x 120 ml) from two different lung segments: with the most (s.A) and with the least (s.B.) extensive involvement evaluated by high resolution computed tomography (HRCT) and in peripheral blood. Examined group consisted of 28 sarcoid patients with homogeneous, regular distribution of nodular opacities in conventional chest X-ray (14 F, 14 M aged 19 54). Twelve patients showed homogeneous distribution of HRCT changes (RD) in lung parenchyma and 16 showed nonhomogeneous distribution of HRCT changes (ND) with domination of pathological changes in upper lobes. Eleven healthy volunteers served as controls. Lymphocytes T gd were estimated by flow-cytometry. In peripheral blood of patients with sarcoidosis the mean value of T gd lymphocytes (4.75%) did not differ from control group (5.3%). In all patients the mean values of T gd percentage in BAL from s.A (1.7 + 1.0%) and in BAL from s.B (2.1 + 1.5%) were significantly lower (p < 0.01) than the mean value in peripheral blood (4.75 + 2.4%) and were significantly lower than mean value of T gd cells in BAL from s.B bis (4.2 + 2.7%). Among subgroups ND and RD we did find any significant differences between values of T gd in BAL form s.A and s.B. Our results suggest minimal role of T gd lymphocytes in sarcoid pathogenesis. PMID- 10808388 TI - [BALF from two different lung segments indicated by high resolution computer tomography (HRCT) in patients with sarcoidosis. III. Correlation between pulmonary function tests and HRCT changes and BAL cell count]. AB - In 28 patients with sarcoidosis patients (14 F, 14 M aged 19-54) lymphocytes subpopulations were estimated in double BAL from two lung segments: with the most (s.A) and with the least (s.B.) extensive involvement estimated by high resolution computed tomography (HRCT). HRCT score for whole lung correlated negatively with DCO (r = 0.46, p < 0.05), D/VA (r = -0.46 p < 0.05), Cstat (r = 0.57, p < 0.05) and Cdyn (r = 0.-057, p < 0.01). HRCT-score for lung segments A and B did not correlate with BAL-cell count and lymphocytes subsets from these segments. The relationship between percentage of lymphocytes HLA-DR in BAL from s.A and d(A-a)O2 (r = 0.38, p < 0.05) and the relationship between absolute number of CD25 in BAL from s.A and DCO (r = -0.38, p < 0.05) were observed. The percentage of lymphocytes in BAL from s.B correlated negatively with D/VA (r = 0.40, p < 0.05) and the percentage of HLA-DR lymphocytes in BAL from s.B. correlated negatively with Cdyn (r = -0.45, p < 0.05). Our results suggests usefulness of HRCT in estimation of sarcoidosis advancement but not in it's activity and indicate the careful interpretation the relationships between BAL results from only one lung segment and pulmonary function parameters. PMID- 10808389 TI - [Comparison of the influence of celiprolol, metoprolol and atenolol on pulmonary ventilation in patients with asthma]. AB - Usage of beta-blocking agents in patients with bronchial asthma is restricted due to their ability to precipitate bronchospasm. Celiprolol beta1-selective beta blocker with associated beta2-agonist activity gives brand new possibilities of treatment with beta-blockers in asthmatics. The aim of the study was to compare the pulmonary effects of single dose of celiprolol (200 mg), atenololol (25 mg), metoprolol (50 mg) and placebo. Ten stable asthmatic patients, aged 21-60 years (mean age 44.1 yrs.) were studied. During four separate visits with 3 days wash out period physical examination, ECG recordings were done and lung function tests (FEV1, FVC, FEF25-75), blood pressure, heart rate and saturation were recorded. All parameters were measured again after 45, 90, 150 and 210 minutes from the beginning of the visit. On the contrary to metoprolol and atenolol single dose of celiprolol and placebo did not significantly affect respiratory functions (FVC- area under curve). There was significant decrease of FEV1 calculated as area under curve (AUC) after application of metoprolol in comparison to celiprolol. There were no significant changes in FEV1 after use of atenolol and celiprolol. Celiprolol ian dose 200 mg can be safely used in asthmatic patients. PMID- 10808390 TI - [Myocardial infarction as a cause of death in patients treated for small cell lung cancer]. AB - The aim of our study was to assess frequency of death from myocardial infarction in patients (pts) treated for small cell lung cancer (SCLC). 33 out of 845 patients treated for SCLC died from myocardial infarction. All patients were smokers. In 6 patients coexisted hypertension, in 2--diabetes and in 5--obesity. Eight patients have had cardiac disease in anamnesis. All patients were treated with one or more number of cardiotoxic drugs as DDP, VCR or VBL, E, MTX and ADR which are able to cause ischemic heart disease or myocardial infarction. Sixteen out of 33 patients have had radiotherapy of lung tumour. Death from myocardial infarction occurred from 0.5 up till 98.5 months from the beginning of start treatment. Eighteen men died from myocardial infarction in the first year of treatment. Risk of death from myocardial infarction was 15 times greater in men with SCLC than in men of the polish population at the same age and at the same time. PMID- 10808391 TI - [Pulmonary thromboembolism as the main or secondary cause of death in patients treated for small cell lung cancer]. AB - The aim of this study was to assess the frequency of pulmonary thromboembolism in small cell lung cancer (SCLC) patients (pts) treated in the Institute of Tuberculosis and Chest Diseases in the years 1980-1994. The second aim was to assess whether the occurrence of thromboembolism is connected with the progression of cancer. Pulmonary thromboembolism was recognised in 17 out of 845 pts (2.01%) treated for small cell lung cancer, that is in 2.01% of pts. The frequency of thromboembolism was 8% among patients in which autopsy was done. Pulmonary thromboembolism occurred in 7 pts during progressive disease but also in 9 with partial or complete regression of SCLC. The potential role of chemo- and radiotherapy in the development of thromboembolism in the last group was discussed. PMID- 10808392 TI - [Tuberculosis of the tongue]. AB - A case of coexisting tongue and pulmonary tuberculosis is presented. A 55 year old man, farmer, an alcohol abuser, suffered from respiratory symptoms infection was admitted to Dental Surgery Department because of carcinoma of tongue suspicion. His tongue was oedematous, stiff with numerous painful ulcerations. Thin-needle biopsy of the tongue revealed granuloma typical for tuberculosis. Chest x-ray examination revealed disseminated lesions and cavernous infiltration in the lungs. Tuberculosis of lungs and tongue was recognised. Antimycobacterial treatment was introduced. 2 weeks later, tuberculous bacilli were cultured. Excellent clinical and mainly-local (tongue) improvement has been achieved. PMID- 10808393 TI - [[A case of primary tracheo-bronchial amyloidosis]. AB - A case of tracheobronchial amyloidosis is described in a 39 year-old man. The diagnosis was established histologically by a bronchoscopic biopsy which revealed a diffuse infiltrative process in the tracheobronchial tree. Due to the symptoms of obstruction of the lower respiratory tract the patient was treated with Prednisone and Melphalan and with partial excision of amyloid deposits by bronchoscopy. Bronchoscopy although carrying a risk of bleeding is the procedure of choice in diagnosis and therapy of such cases. PMID- 10808394 TI - 21st Annual meeting of the Society for Clinical Trials. Toronto, Canada, April 16 19, 2000. Abstracts. PMID- 10808395 TI - 1st European Congress of Andrology. L'Aquila, Italy, 24-27 March 2000. Abstracts. PMID- 10808396 TI - XXXIV Angiologic Meeting of The French Language. Paris, France. 15-17 March 2000. Abstracts. PMID- 10808397 TI - American Society for Laser Medicine and Surgery 20th annual meeting. Reno, Nevada, USA. April 7-9, 2000. Abstracts. PMID- 10808398 TI - German Society for Experimental and Clinical Pharmacology and Toxicology 41st Spring Meeting. Mainz, Germany, 21-23 March 2000. Abstracts. PMID- 10808399 TI - Symposium on RNA Biology. III. RNA, Tool and Target. Research Triangle Park, North Carolina, USA. October 15-17, 1999. Proceedings. PMID- 10808400 TI - Expanding role of monoclonal antibodies in the treatment of hematologic malignancies. Highlights from the 41st annual meeting of the American Society of Hematology. New Orleans, Louisiana, USA. December 3-7, 1999. Abstracts. PMID- 10808401 TI - German Society of Physiology 79th annual meeting. Ulm, 19-22 March 2000. Abstracts. PMID- 10808402 TI - The 4th Italian Society of Toxicology (SITOX) Symposium on Molecular and Cellular Toxicology and 3rd National Congress of Celltox-Italian Association of In Vitro Toxicology. Salerno, Italy, 5-7 November 1997. Abstracts. PMID- 10808403 TI - 10th Biennial Winter Workshop on Schizophrenia. Davos, Switzerland, February 5 11, 2000. Abstracts. PMID- 10808404 TI - [Intracellular proteolysis: signals of selective protein degradation]. AB - Selective proteolysis is one of the mechanisms for the maintenance of cell homeostasis via rapid degradation of defective polypeptides and certain short lived regulatory proteins. In prokaryotic cells, high-molecular-mass oligomeric ATP-dependent proteases are responsible for selective protein degradation. In eukaryotes, most polypeptides are attacked by the multicatalytic 26S proteasome, and the degradation of the majority of substrates involves their preliminary modification with the protein ubiquitin. The proteins undergoing the selective proteolysis often contain specific degradation signals necessary for their recognition by the corresponding proteases. PMID- 10808405 TI - [Indole derivatives in vegetables of the family Cruciferae]. AB - The chemical background of the biological activities of vegetables of the Cruciferae family is considered. These vegetables contain alkaloids of the glucobrassicin group that are decomposed by the enzyme myrosinase (thioglucosidase, EC 3.2.3.1) released upon damage to the plant cells. This results in several indole derivatives, with ascorbigen and indole-3-carbinol predominating. In the gastrointestinal tract, these compounds form 5H,11H indolo[3,2-b]carbazole, natural ligand of the aromatic hydrocarbon receptor (Ah receptor) and a functional analogue of 2,3,7,8-tetrachlorodibenzo-p-dioxin, a dangerous xenobiotic. The indolocarbazole-Ah receptor complex activates the gene of CYP1A1, an isoenzyme of cytochrome P450-dependent monoamine oxidase, which enhances the 2-hydroxylation (inactivation) of estrogens. In its turn, the resulting lowered level of estrogens inhibits the growth of hormone-dependent tumors or prevents their appearance. The mechanism of xenobiotic inactivation, underlying the anticarcinogenic action of food products including vegetables of Cruciferae family and some homogeneous indole compounds, is similar. Some other effects of nutrient indole compounds, e.g., the inhibition of expression of the cyclin-dependent kinase 6 (CDK6) by indole-3-carbinol that leads to the cell cycle arrest in G1 phase, are also considered. Analysis of the biological effects of the Cruciferae diet has helped start clinical studies of indole-3-carbinol as an antitumor and anticarcinogenic remedy for patients with a high risk of tumor diseases. PMID- 10808406 TI - [the synthesis of diporphyrin systems on the basis of tetraphenylporphyrin derivatives bridged with peptide spacers]. AB - Diporphyrin systems based on tetraphenylporphyrin derivatives bridged with dipeptide or tripeptide spacers containing Gly and Phe residues were synthesized, and their physicochemical properties were studied. PMID- 10808407 TI - [The synthesis of a cobalt(II) tetracarboxyphthalocyanine- deoxyribooligonucleotide conjugate as a reagent for the directed DNA modification]. AB - The cobalt(II) tetracarboxyphthalocyanine-deoxyribonucleotide pd(TCTTCCCA) conjugate was synthesized. The phthalocyanine N-succinimide ester prepared from phthalocyanine using DCC was mixed in DMF with an aqueous solution of the oligonucleotide bearing a 1,3-diaminopropane linker at the 5'-phosphate. The resulting conjugate was tested in the intraduplex reaction with target 14-mer and 22-mer oligonucleotides containing conjugate-complementary sequences. In the presence of O2 and a thiol (2-mercaptoethanol or DTT) as a coupled reducer or H2O2, sequence-specific DNA modification was observed that caused the cleavage of the target upon treatment with piperidine. PMID- 10808408 TI - [The synthesis of galactopyranosyl-substituted derivatives of pheophorbide]. AB - New pheophorbide and pyropheophorbide derivatives containing carbohydrate fragments, derivatives of galactopyranose, were synthesized. Galactopyranosylpheophorbide and galactopyranosylpyropheophorbide with free hydroxyl groups were found to be water-soluble and useful as sensitizers in photodynamic therapy of cancer. PMID- 10808409 TI - [Modeling of peptides and proteins in membrane environment. I. A solvation model mimicking a lipid bilayer]. AB - A theoretical solvation model of peptides and proteins that mimics the heterogeneous membrane-water system was proposed. Our approach is based on the combined use of atomic parameters of solvation for water and hydrocarbons, which approximates the hydrated polar groups and acyl chains of lipids, respectively. This model was tested in simulations of several peptides: a nonpolar 20-mer polyleucine, a hydrophobic peptide with terminal polar groups, and a strongly amphiphilic peptide. The conformational space of the peptides in the presence of the membrane was studied by the Monte Carlo method. Unlike a polar solvent and vacuum, the membrane-like environment was shown to stabilize the alpha-helical conformation: low-energy structures have a helicity index of 100% in all cases. At the same time, the energetically most favorable orientations of the peptides relative to the membrane depend on their hydrophobic properties: nonpolar polyleucine is entirely immersed in the bilayer and the hydrophobic peptide with polar groups at the termini adopts a transbilayer orientation, whereas the amphiphilic peptide lies at the interface parallel to the membrane plane. The results of the simulations agree well with the available experimental data for these systems. In the following communications of this series, we plan to describe applications of the solvation model to membrane-bound proteins and peptides with biologically important functional activities. PMID- 10808410 TI - [Reactive derivatives of oligonucleotide phosphorothioate analogues. IV.Site directed modification of nucleic acids in intramolecular alkylation of phosphorothioate groups]. AB - Alkylation of the 22-mer DNA target pTGCCTGGAGCTGCTTGATGCCC (I) by oligodeoxynucleotide phosphorothioate derivatives (PTAO) GpsCpsApsTpsCpsApsApsGpsCpsApsGpsCpN(CH3)CH2(RCl) (II-PS) and (RCl)CH2N(CH3)pGpsCpsApsTpsCpsApsApsGpsCpsApsGpsC (III-PS) bearing a residue of an aromatic analogue of nitrogen lost (RCl = C6H4N(CH3)(CH2CH2Cl) at the 3'- or 5'-end was studied. It was shown that the internucleotide phosphorothioate bonds do not affect the regiospecificity of the target modification. The maximum degree of the target modification (at t-->infinity) at 20 degrees C was about 25% for both (II-PS) and (III-PS). The use of GCATCAAGCAGCpN(CH3)CH2(RCl) (II-PO), containing internucleotide phosphodiester bonds, under the same conditions gave about 65% of the modified DNA. Kinetics of the PTAO-induced complementarily addressed nucleic acid (NA) modification was analyzed. The rate constants of the reaction of the intermediate reactive ethylenimmonium ion with phosphorothioate groups of the reagents were evaluated both in solution and in duplex. The intramolecular alkylation of phosphorothioate groups considerably affected the DNA target modification by decreasing the effectiveness of the modification in a wide range of temperatures and changing the temperature dependence of the modification from a bell-like to an S-like profile. It was concluded that, in the course of the modification, the PTAO phosphorothioate groups are intramolecularly alkylated both in solution and in the complementary NA target-oligonucleotide duplex. PMID- 10808411 TI - [A photoreactive analogue of 2'3'-dideoxyuridine 5'-triphosphate: preparation and use for photoaffinity modification of human replication protein A]. AB - A new reagent for photoaffinity modification of biopolymers, 5-[E-N-(2-nitro-5 azidobenzoyl)-3-amino-1-propen-1-yl]-2',3'-dideoxyuridine 5'-triphosphate (NAB ddUTP), was synthesized. Like a similar derivative of 2'-deoxyuridine 5' triphosphate (NAB-dUTP), it was shown to be able to effectively substitute for dTTP in the synthesis of DNA catalyzed by eukaryotic DNA polymerase beta and to terminate DNA synthesis. A 5'-32P-labeled primer with a photoreactive group at the 3'-terminus was derived from NAB-ddUTP and used for photoaffinity labeling of the human replication protein A (RPA). The covalent attachment of RPA p32 and p70 subunits to the labeled primers was demonstrated. NAB-ddUTP is a promising tool for studying the interaction of proteins of the replicative complex with NA in cellular extracts and living cells during the termination of DNA synthesis. PMID- 10808412 TI - [Inhibition of horseradish peroxidase by N-ethylamide of o-sulfobenzoylacetic acid]. AB - The carboxylic groups of horseradish peroxidase were modified by 1-cyclohexyl-3 (2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate by the Koshland method. The catalytic properties of the native and modified peroxidase were studied in the presence of N-ethylamide of o-sulfobenzoylacetic acid (EASBA) at pH 5.0-7.5. In the oxidation of o-dianisidine, EASBA is a competitive inhibitor of the carbidiimide-modified peroxidase, and it increases both K(m) and Vm in the case of the native enzyme. These data show that at least one of the carboxylic groups modified with carbodiimide is located at the area of the peroxidase active site. PMID- 10808413 TI - 3D imaging and its derivatives in clinical research and practice. PMID- 10808414 TI - Perceptual gaps in understanding patient expectations for health care service quality. AB - This article assesses how well physicians, health administrators, patient-contact employees, and especially medical and nursing students understand patient expectations for service quality as measured by the SERVQUAL scale. Using a cross sectional research design and discriminant analysis, it was found that health administrators were most likely to accurately estimate the service expectations of patients, while medical and nursing students were most likely to underestimate them. PMID- 10808415 TI - The diffusion of information technology among health maintenance organizations. AB - This article examines the information technology functions, staffing and cost, services provided, and advanced technologies among health maintenance organizations (HMOs) using a national sample of HMOs from mid-1995. HMOs have a well-developed capability to use data from administrative functions, such as claims processing. Nationally affiliated HMOs and HMOs in markets with greater HMO penetration support more IT functions. Relatively little work has been completed integrating clinical with administrative systems. PMID- 10808416 TI - Improving the public sector: can reengineering identify how to boost efficiency and effectiveness at a VA medical center? AB - Reengineering is a widespread management technique, but few evaluations of its application in health care, especially in public sector organizations, have been reported. We conducted a reengineering analysis to determine if the method could identify how to improve the efficiency and effectiveness of a VA medical center's primary care delivery system. We found that the reengineering method appears applicable in the institutional context of a public sector teaching hospital. PMID- 10808417 TI - Mission statements in Canadian not-for-profit hospitals: does process matter? AB - Mission statements abound in health care organizations. And much is written on what they should contain. But, the process of creating and implementing mission statements in health care organizations has received virtually no attention in the literature. This article presents findings from a research study that sought to determine whether or not a relationship existed between selected mission process characteristics and various measures of a hospital's performance. PMID- 10808418 TI - A multi-level performance appraisal tool: transition from the traditional to a CQI approach. AB - The commitment of many health care organizations to CQI calls for the introduction of tools, principles, and procedures to ensure that the performance improvement in all parts of the organization is initiated. In order to determine whether the desired performance standards are indeed being achieved on all organizational levels, it is necessary to apply a holistic performance appraisal tool that would allow for an assessment of the company's progress from multiple dimensions. This article details the design of the multilevel appraisal model, explicates where the evaluation errors may occur, and presents managerial implications for successful implementation of this evaluation tool. PMID- 10808419 TI - Disaster recovery planning by HMOs: theoretical insights. AB - HMOs are becoming increasingly reliant on health management information systems (HMISs) for their effective functioning, competitive viability, and survival. Because of this critical dependence, HMOs must use disaster recovery planning (DRP) to safeguard their HMIS assets from natural as well as man-made disasters. This article purposes a theoretical framework, based on the theory of organizational innovation, to explain the factors involved in the adoption of DRP by HMOs. PMID- 10808420 TI - The corporate social responsiveness orientation of hospital directors: does occupational background make a difference? AB - The results of a survey of 184 directors from 15 hospitals are analyzed to determine their corporate social responsiveness orientation. They indicate that board members whose occupational background is not in health care exhibit greater concern for economic performance and the legal component of corporate responsibility than their counterparts whose occupational background is in health care. No significant differences between the two groups were observed with respect to the ethical and discretionary dimensions of corporate social responsibility. Some explanations as well as limited generalizations and implications are developed. PMID- 10808421 TI - Hospital selection by managed care insurers. AB - This article focuses on which factors influence the ability of hospitals to attract managed care patients. Data for the analysis were obtained from a national sample of 235 acute care hospitals in 1996. The results indicate that the costs of inpatient care, participation in an integrated delivery system network, and hospital size were the critical factors in determining the ability of hospitals to obtain managed care business. PMID- 10808422 TI - [Usefulness of rapid bedside cardiac troponin T assay for the diagnosis of acute myocardial infarction]. AB - Cardiac troponin T(cTnT) is one of the most myocardial-specific markers for the diagnosis of acute myocardial infarction(AMI). Recently, the rapid bedside cTnT assay(Trop T rapid assay sensitive version), which can provide qualitative determinations within 15 min, has been developed for the emergency clinical setting. To evaluate the usefulness of rapid bedside cTnT assay, we performed the Trop T test and measured serum levels of myoglobin(Mb), creatine kinase MB isoenzyme(CK-MB) and cTnT in 256 consecutive emergency patients with suspected AMI(65 found to have AMI and 191 without AMI). The diagnostic sensitivities for AMI of Trop T, Mb and CK-MB measurements were 66%, 92% and 52%, respectively, whereas the specificities were 80%, 18% and 74%, respectively. The diagnostic accuracy for AMI of Trop T(77%) was significantly higher than that of Mb(37%, p < 0.001) and CK-MB(69%, p < 0.05). The sensitivity for AMI of Mb(86%) was significantly(p < 0.001) higher than that of Trop T (31%) and CK-MB(31%) in patients admitted < or = 3 hr after the onset of AMI. In contrast, the sensitivities of Trop T(80% and 100%) in patients admitted at 3-6 hr and > 6 hr showed no significant differences from those of Mb(100% and 96%). Furthermore, Trop T in patients admitted > 6 hr had significantly(p < 0.01) higher sensitivity compared with CK-MB(69%). The mortality rate in the non-AMI group during hospitalization in patients with positive Trop T test(39%) was significantly(p < 0.001) higher than that in patients with negative test(9%). When the positive Trop T test was regarded as > or = 0.10 ng/ml of serum cTnT, Trop T test had the best concordance of 92% with a quantitative of cTnT assay. PMID- 10808423 TI - [Prediction of restenosis after percutaneous transluminal coronary angioplasty using coronary flow reserve. Kansai Doppler Guide Wire Study Group]. AB - This multi-center prospective study attempted to predict restenosis after percutaneous transluminal coronary angioplasty(PTCA) using coronary flow reserve. Intracoronary blood flow velocity was measured in 47 patients(37 males, 10 females, mean age 66 +/- 9 years) with a Doppler guide wire, following successful PTCA. Twenty-four patients had prior myocardial infarction. After successful PTCA, a Doppler guide wire was placed at the distal portion of the target lesion, and coronary blood flow velocity was measured before and during intravenous administration of adenosine triphosphate. Follow-up coronary angiography was performed 154 +/- 69 days after PTCA, and the diameter stenosis of the target lesion was measured using quantitative coronary angiography. Follow-up angiography showed restenosis in 13 patients(28%). Sensitivity and specificity for predicting restenosis were low(50%, 45%, respectively) with a post-PTCA% diameter stenosis cut-off point of 27%. Sensitivity and specificity were 67% and 61% with a minimal lumen diameter cut-off of 1.8 mm. The reference coronary artery diameter(cut-off point 2.5 mm) was better for predicting restenosis(sensitivity 78% and specificity 76%). Sensitivity and specificity were 62% and 67%, respectively, using coronary flow reserve(cut-off point 2.0). The restenosis rate of patients with a reference diameter of more than 2.5 mm was 10%, but 54% for those with less than 2.5 mm(p < 0.05). In patients with a reference diameter of less than 2.5 mm, coronary flow reserve was useful for predicting restenosis(cut-off point 1.9, sensitivity 71% and specificity 83%). Coronary flow reserve is useful for predicting restenosis after PTCA, when combined with reference coronary artery diameter. PMID- 10808424 TI - [Coronary flow after successful angioplasty in patients with acute myocardial infarction: comparison between coronary flow reserve by nicorandil and papaverine]. AB - In order to evaluate coronary flow response to 2 different vasodilators, nicorandil and papaverine, in patients with myocardial infarction, we measured coronary flow reserve using a Doppler guide wire in infarct-related and non infarct-related arteries. The study group consisted of 28 patients with first acute myocardial infarction 3 weeks after successful coronary angioplasty within 6 hr after symptom onset. Twelve patients with atypical chest pain served as the control group. Coronary flow reserve induced by intracoronary papaverine(12 mg) was lower in infarct-related arteries than in non infarct-related arteries, but there were no differences in coronary flow reserve induced by intracoronary nicorandil(1 mg) between infarct-related and non infarct-related arteries. Coronary flow reserve induced by nicorandil was lower than that by papaverine in non infarct-related arteries and the control group. However, there were no differences between coronary flow reserve induced by nicorandil and papaverine in infarct-related arteries. Vasodilatory response induced by nicorandil was relatively preserved in infarct-related arteries compared with papaverine. These results suggest that impairment of coronary microvascular response in infarct myocardium varies in the different sites acted on by different vasodilator agents. PMID- 10808425 TI - Correlation between interventricular septal motion and left ventricular systolic diastolic function in patients with left bundle branch block. AB - The echocardiographic correlation between abnormal interventricular septal motion including systolic paradoxical, flat motion and early diastolic notch and ejection fraction, mean ventricular circumferential fiber shortening and early diastolic mitral filling velocity was studied in 46 patients with complete left bundle branch block. Systolic normal interventricular septal motion was used as the control. Ejection fraction was significantly smaller in the paradoxical(0.44 +/- 0.13, p < 0.02) and flat motion groups(0.38 +/- 0.09, p < 0.001) than in the normal group(0.54 +/- 0.1). Mean ventricular circumferential fiber shortening was significantly smaller in the flat motion group than in the normal group(0.72 +/- 0.19 vs 0.99 +/- 0.23 circ/sec, p < 0.002). The size of the notch was significantly smaller in the flat motion group than in the normal group (2.3 +/- 0.2 vs 4.8 +/- 0.8 mm, p < 0.0001). The deceleration rate of the notch was significantly slower in the paradoxical and flat motion groups than in the normal group(37.3 +/- 12.2, 31.3 +/- 8.1 vs 69.1 +/- 2.5 cm/sec, p < 0.0001). Early diastolic mitral filling velocity was slower in the flat motion group than in the other 2 groups. In conclusion, systolic flat interventricular septal motion showed more severe disturbances of left ventricular systolic and diastolic function than paradoxical interventricular septal motion in patients with complete left bundle branch block. PMID- 10808426 TI - [Multi-vasospastic angina refractory to medical therapy caused by hyperthyroid stage of chronic thyroiditis and hypereosinophilia: a case report]. AB - A 49-year-old woman with bronchial asthma was followed up at our hospital. After 3 years, she experienced an attack of chest pain with ST elevation in the precordal leads of electrocardiography. After admission, the chest pain and ST elevation disappeared, but the chest pain recurred after 6 days. Coronary angiography revealed no significant stenosis in the coronary arteries. After discharge, she had the chest pain repeatedly. ST elevation in the II, III, aVF leads was recorded. The diagnosis was coronary multispasm. The chest pain was refractory to medical therapy. Hypereosinophilia developed and bronchial asthma worsened. After steroid administration, the angina and bronchial asthma ceased. She has lost about 15 kg during 1 year. Laboratory data revealed low thyroid stimulating hormone, high thyroid hormone, positive thyroglobulin antibody, and negative thyroid-stimulating hormone receptor antibody. The diagnosis was chronic thyroiditis. The multi-vasospastic angina refractory to medical therapy was caused by the hyperthyroid stage of chronic thyroiditis and hypereosinophilia. PMID- 10808427 TI - [Efficacy of continuous intravenous drip infusion of disopyramide in hypertrophic obstructive cardiomyopathy during cardiogenic shock: a case report]. AB - A 54-year-old woman was admitted to our hospital complaining of dyspnea due to hypertrophic obstructive cardiomyopathy. On admission, she was treated with 4 antiarrhythmic drugs and 2 beta-blockers. After 4 of these 6 drugs were withdrawn, the left ventricular outflow pressure gradient markedly increased and then she fell into cardiogenic shock. Therefore, disopyramide(600 mg/day) was administered by continuous intravenous drip infusion to reduce the left ventricular outflow pressure gradient. After intravenous administration of disopyramide, the left ventricular outflow pressure gradient markedly decreased from 100 to 16 mmHg and the cardiogenic shock could be improved. Continuous intravenous drip infusion of disopyramide is effective for the treatment of cardiogenic shock due to severe left ventricular outflow obstruction in patients with hypertrophic obstructive cardiomyopathy. PMID- 10808429 TI - [Cardiovascular imaging in-a-month. An 11-year-old boy with refractory congestive heart failure]. PMID- 10808428 TI - [Transmural heterogeneity of the left ventricular wall: subendocardial layer and subepicardial layer]. AB - The myocardium of the left ventricular wall is not homogeneous, but demonstrates transmural heterogeneity in myocardial blood flow, myocardial metabolism, and contraction and relaxation dynamics. Reimer and colleagues recognized that irreversible injury of the ischemic myocardium develops as a transmural wavefront, occurring first in the subendocardial myocardium, and with longer periods of ischemia, the wavefront of necrosis moves from the subendocardial zone across the wall to progressively involve more of the transmural thickness of the ventricular wall, ultimately becoming nearly transmural. This phenomenon was named the "wavefront phenomenon", and is the morphological counterpart of the transmural heterogeneity of the metabolism and blood flow. Autoregulation of myocardial blood flow is accomplished by changes in intramyocardial vascular resistance and intramyocardial pressure. It is more difficult to maintain the autoregulation in the subendocardial myocardium because contraction is greater, oxygen demand is greater, and myocardial pressure is higher in the subendocardium than in the subepicardial layer. In the normal myocardium, contraction is greater in the subendocardial layer, as is wall stress, accounting for the higher subendocardial energy requirements. Consistent with these findings, higher rates of metabolic activity and greater oxygen extraction are found in this region. As a consequence, ischemia becomes more severe and myocardial cells undergo necrosis first in the subendocardium. Under normal conditions, production and utilization of high-energy phosphates [adenosine triphosphate(ATP) and creatine phosphate] in the subendocardial myocardium are more active than in the subepicardial myocardium, but decline more easily in the subendocardium during ischemia, which induces the subendocardial ischemic injury. Lower production of Ca(2+)-ATPase in the subendocardium might also contribute to the subendocardial injury. Wavefront necrosis starts from the subendocardium, but the production of high-energy phosphates in the subepicardium is known to increase and compensate for the reduction in high-energy phosphate production in the subendocardium. Animal experiments have shown that systolic thickening of the endocardial half of the ventricular wall is double that in the epicardial half. Today, this can be confirmed in humans with the tissue Doppler tracking method which is completely noninvasive. Furthermore, the subepicardial half of the ventricular wall is known to compensate for the decreased systolic thickening of the subendocardial half in the case of subendocardial injury, which is called vertical compensation and is the mechanical counterpart of the concept of metabolic compensation. Many new technologies, including the tissue Doppler tracking method, magnetic resonance imaging tagging, and myocardial contrast echocardiography, will give more accurate information about the myocardial heterogeneity of layer-by-layer motion and blood flow, and will contribute to early detection and quantitative estimation of ischemia and other diseases of which the main lesion is in the subendocardium. PMID- 10808430 TI - [Cardiovascular imaging in-a-month. Large blinded cavity in the right ventricle detected by electrocardiogram-gated cardiac electron-beam computed tomography]. PMID- 10808431 TI - Synthesis of fluorous fullerene adducts: reversible solubilization of fullerenes in perfluorinated solvents AB - Fullerene (C60) Diels-Alder adducts with perfluoroalkylated 1,3-cyclopentadiene (1a,b) were synthesized and studied. The perfluoroalkylated cyclopentadiene was found to be less reactive toward C60 than cyclopentadiene itself, possibly because of the electron-withdrawing effect of the side chain. Because of the same effect, the temperature of the retro-Diels-Alder reaction for the fluorinated adducts was lower (70 degrees C) than the reported value (95 degrees C) for the cyclopentadiene adducts of C60. Higher adducts of the fluorous diene and C60 were found to be soluble in perfluorohexane and to show visible partitioning between organic (toluene) and fluorous phases. Also, the Diels-Alder addition of the fluorous diene was accompanied by extensive oxidation of the fullerene core, as revealed by MALDI-TOF data. PMID- 10808432 TI - Selectively monomodified cyclodextrins. Synthetic strategies AB - Monomodifications of cyclodextrins to give selectively 2-, 3-, or 6-substituted product is a challenging task because of the number of hydroxyl groups that can potentially react with the incoming reagent. The principles and the methods involved in manipulations of the differences in the chemistry of these hydroxyl groups to control the outcome of an electrophilic reaction with them to produce monoalkylated (ether-linkaged) cyclodextrin derivatives are discussed and illustrated. PMID- 10808433 TI - Theoretical study of the structure-property relationship in phosphole monomers AB - Use of the DFT method with the B3LYP functional offers an efficient way for determining geometries of phosphole structures. It also provides inversion barriers in good agreement with experiment. The molecular orbital MP2 method is better suited to calculate the relative stability of the considered isomers. Combination of geometric (Julg index) and magnetic (NICS) criteria leads to an interesting approach to analyze the pi-electron delocalization and/or aromaticity in conjugated heterocyclic systems and can be used for investigation of much larger oligomers. It thus allows us to confirm that planarizing the phosphorus atom in phosphole monomers leads to a greater conjugation over the butadienic pi system and that substituents, characterized by a strongly pronounced pi-system, exhibit a large extent of conjugation with the pi-diene moiety of the heterocyclic system. In push-pull systems such as 2-BH2-5-NH2-1H-phosphole, the electron delocalization along the pi-diene system is even more pronounced than in phospholes with strongly pronounced pi-systems. The employed approach is also used to analyze the relationship between electron conjugation and the phosphorus inversion barrier. More relevant for chemical applications is the result that the computational study provides a method of fine-tuning of phosphole building blocks, which may enable us to obtain higher pi-conjugation along the polymer backbone. PMID- 10808434 TI - Diastereoselective addition of amines to vinyl sulfone modified carbohydrates: a highly flexible methodology for the synthesis of new classes of deoxyaminosugars AB - Methyl 2,3-dideoxy-4,6-O-(phenylmethylene)-3-C-phenylsulfonyl-alpha- D-erythro hex-2-enopyranoside (5 alpha) and methyl 2,3-dideoxy-4,6-O-(phenylmethylene)-3-C phenylsulfonyl- beta-D-erythro-hex-2-enopyranoside (5 beta) have been subjected to Michael addition reaction with various amines to develop a new methodology for the synthesis of new classes of aminosugars. Compound 5 alpha reacted with primary amines to generate gluco- derivatives, but secondary amines produced both gluco- (major) and manno- (minor) isomers. Compound 5 beta, on the other hand, produced only gluco- isomers with both primary and secondary amines. The stereochemical course of addition of some of the amines to 5 alpha and 5 beta are significantly different from that of the addition of amines to 3 nitroenopyranoses. The present route to the syntheses of various aminosugars with gluco- configurations from 5 alpha and 5 beta constitutes a novel method for the introduction of N-monoalkylated and N,N-dialkylated amines to the C-2 carbon of pyranoses in equatorial configurations. PMID- 10808435 TI - Total synthesis of (+/-)-aspidospermidine. AB - (+/-)-Aspidospermidine (1) has been synthesized from readily available methyl 3 ethyl-2-oxocylo-pentanecarboxylate (17) in 5.9% yield over 13 steps. The key step of the synthesis is an intramolecular cascade reaction that simultaneously forms the B, C, and D rings of 1. A high-yielding method of closing the remaining E ring is also described. PMID- 10808436 TI - Synthesis of ring D modified morphinan systems via ring expansion of a key codeine intermediate AB - Ring expansion and ring fragmentation products were synthesized from pentacyclic ketones 3 and 4 via Baeyer-Villiger oxidation. Oxidation of 3 with m chloroperbenzoic acid afforded delta-lactone 9 which was transformed to enone 12 en route to the C6 inverted ether 14. Analogous oxidation of 4 gave delta-lactone 18. Opening of the lactone ring of 14 and 18 led to seco structures which, in the case of 19, underwent reannulation with incorporation of nitrogen to yield the hydroxylactam 21. PMID- 10808437 TI - Hydrolytic reactions of the phosphorodithioate analogue of uridylyl(3',5')uridine: kinetics and mechanisms for the cleavage, desulfurization, and isomerization of the internucleosidic linkage AB - The hydrolytic reactions of the phosphorodithioate analogue of uridylyl(3',5')uridine [3',5'-Up(s)2U] were followed by HPLC over a wide pH range at 363.2 K. Under acidic and neutral conditions, three reactions compete: (i) desulfurization to a mixture of the (Rp)- and (Sp)-diastereomers of the corresponding 3',5'- and 2',5'-phosphoromonothioates [3',5'- and 2',5'-Up(s)U], which are subsequently desulfurized to a mixture of uridylyl(3',5')- and (2',5')uridine [3',5'- and 2',5'-UpU], (ii) isomerization to 2',5'-Up(s)2U, and (iii) cleavage to uridine, in all likelihood via a 2',3'-cyclic phosphorodithioate (2',3'-cUMPS2). Under alkaline conditions (pH > 8), only a hydroxide ion catalyzed hydrolysis to uridine via 2',3'-cUMPS2 takes place. At pH 3-7, all three reactions are pH-independent, the desulfurization being approximately 1 order of magnitude faster than the cleavage and isomerization. At pH < 3, all the reactions are hydronium ion catalyzed. On going to very acidic solutions, the cleavage gradually takes over the desulfurization and isomerization. Accordingly, the cleavage overwhelmingly predominates at pH < 0. The overall hydrolytic stability of 3',5'-Up(s)2U is comparable to that of (Sp)- and (Rp)-3',5'-Up(s)U (and to that of 3',5'-UpU, except at pH < 2). The rate of the hydroxide ion catalyzed hydrolysis of 3',5'-Up(s)2U is 37% and 53% of that of (Sp)- and (Rp)-3',5'-Up(s)U, respectively. The reactions, however, differ with the respect of the product accumulation. While the phosphoromonothioates produce a mixture of 2'- and 3'-thiophosphates as stable products, 3',5'-Up(s)2U is hydrolyzed to uridine without accumulation of the corresponding dithiophosphates. At pH < 3, where the hydrolysis is hydronium ion catalyzed, the kinetic thio effect of the second thio substitution is small: under very acidic conditions (Ho -0.69), (Sp)-3',5'-Up(s)U reacts 1.6 times as fast as 3',5'-Up(s)2U, but the reactivity difference decreases on going to less acidic solutions. In summary, the hydrolytic stability of 3',5'-Up(s)2U closely resembles that of the corresponding phosphoromonothioate. While replacing one of the nonbridging phosphate oxygens of 3',5'-UpU with sulfur stabilizes the phosphodiester bond under acidic conditions by more than 1 order of magnitude, the replacement of the remaining nonbridging oxygen has only a minor influence on the overall hydrolytic stability. PMID- 10808438 TI - Assignment of the absolute configuration of alpha-chiral carboxylic acids by 1H NMR spectroscopy AB - The prediction of the absolute configuration of alpha-chiral carboxylic acids from the 1H NMR spectra of their esters with (R)- and (S)-ethyl 2-hydroxy-2-(9 anthryl) acetate [(R)- and (S)-9-AHA, 5] is discussed. Low-temperature NMR experiments, MM, semiempirical, and aromatic shielding effect calculations allowed the identification of the main conformers and showed that, in all esters studied, conformer ap is the most stable. A simple model for the assignment of the absolute configuration from NMR data is presented, and its reliability is corroborated with acids 6-31 of known absolute configuration. In addition to 5, other auxiliary reagents with open (32-38) and cyclic (39-42) structures have also been studied. trans-(+)- and (-)-2-phenyl-1-cyclohexanol (41) was found to be particularly efficient and produced delta delta RS values similar to those of 5. PMID- 10808439 TI - Synthesis of glycophostones: cyclic phosphonate analogues of biologically relevant sugars AB - Analogues of L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-mannosamine, and N acetyl neuraminic acid in which the anomeric carbon atom was replaced by a phosphonyl group (phostones or cyclic phosphonates) were synthesized by stereocontrolled methods relying on the Abramov reaction. PMID- 10808440 TI - Kinetics of two pathways in peroxyoxalate chemiluminescence AB - It has been shown that 1,1'-oxalyldiimidazole (ODI) is formed as an intermediate in the imidazole-catalyzed reaction of oxalate esters with hydrogen peroxide. Therefore, the kinetics of the chemiluminescence reaction of 1,1' oxalyldiimidazole (ODI) with hydrogen peroxide in the presence of a fluorophore was investigated in order to further elucidate the mechanism of the peroxyoxalate chemiluminescence reaction. The effects of concentrations of ODI, hydrogen peroxide, imidazole (ImH), the general-base catalysts lutidine and collidine, and temperature on the chemiluminescence profile and relative quantum efficiency in the solvent acetonitrile were determined using the stopped-flow technique. Pseudo first-order rate constant measurements were made for concentrations of either H2O2 or ODI in large excess. All of the reaction kinetics are consistent with a mechanism in which the reaction is initiated by a base-catalyzed substitution of hydrogen peroxide for imidazole in ODI to form an imidazoyl peracid (Im(CO)2OOH). In the presence of a large excess of H2O2, this intermediate rapidly decays with both a zero- and first-order dependence on the H2O2 concentration. It is proposed that the zero-order process reflects a cyclization of this intermediate to form a species capable of exciting a fluorophore via the "chemically initiated electron exchange mechanism" (CIEEL), while the first-order process results from the substitution of an additional molecule of hydrogen peroxide to the imidazoyl peracid to form dihydroperoxyoxalate, reducing the observed quantum yield. Under conditions of a large excess of ODI, the reaction is more than 1 order of magnitude more efficient at producing light, and the quantum yield increases linearly with increasing ODI concentration. Again, it is proposed that the slow initiating step of the reaction involves the substitution of H2O2 for imidazole to form the imidazoyl peracid. This intermediate may decay by either cyclization or by reaction with another ODI molecule to form a cyclic peroxide that is much more efficient at energy transfer with the fluorophore. The reaction kinetics clearly distinguishes two separate pathways for the chemiluminescent reaction. PMID- 10808441 TI - A one-pot bicycloannulation method for the synthesis of tetrahydroisoquinoline systems. AB - A highly effective method for the synthesis of the core indolo[2,3 alpha]quinolizidine skeleton found in yohimbine is described. The reaction of N monosubstituted thioamides with bromoalkenoyl chlorides furnishes thioisomunchnones as transient 1,3-dipoles that undergo ready intramolecular cycloaddition across the tethered pi-bond to give thio-bicycloannulated products in a one-pot operation. The stereochemical outcome of the intramolecular reaction is the consequence of an endo cycloaddition of the neighboring pi-bond across the transient thioisomunchnone dipole. A major limitation of the method is that when a hydrogen is present in the alpha-position of the thioamide the initially formed thio-N-acyliminium ion undergoes proton loss to produce a S,N-ketene acetal at a faster rate than dipole formation. Treatment of tetrahydro-beta-carboline-1 thione with 2-bromooct-7-enoyl chloride followed by reductive removal of sulfur from the cycloadduct resulted in the formation of (+/-)-alloyohimbanone. Attempts to cycloadd the thioisomunchnone dipole across several nucleophilic pi-bonds failed, and instead, products derived from cyclization of the pi-bond onto the initially formed thio-N-acyliminium ion were formed. The resulting N,S-ketals were further converted into several tetrahydroisoquinoline alkaloids in good yield. PMID- 10808442 TI - A pericyclic cascade to the stereocontrolled synthesis of 9-cis-retinoids AB - A domino reaction that is pericyclic in nature is thought to be triggered upon treatment of alkenynol 10 with arylsulfenyl chlorides. The process comprises an ordered sequence of sigmatropic rearrangements: a reversible [2,3]-allyl sulfenate to allyl sulfoxide shift, followed by a [2,3]-propargyl sulfenate to allenyl sulfoxide rearrangement, and last a stereodifferentiating [1,5] sigmatropic hydrogen migration leading to polyene 13. The occurrence of the C7 to C11 hydrogen migration has been demonstrated by labeling experiments. The double diastereoselection of the [1,5]-sigmatropic hydrogen shift to afford a single isomer of the final polyene 13 is thought to arise from a combination of the electronic effect of the sulfoxide at one terminus, and the steric effect imparted by the bulky trimethylcyclohexenyl substituent at the other terminus. The overall process thus constitutes a stereoselective synthesis of an E,Z,Z triene fragment from an alkenynol and, in particular, a retinoid with the 7E,9Z,11Z,13E configuration on the conjugated polyenic side chain. Application of this method to the synthesis of retinoids, including labeled analogues, is straightforward. PMID- 10808443 TI - Acylthioketene-thioacylketene-thiet-2-one rearrangements AB - Flash vacuum thermolysis (FVT) of 6-aryl-1,3-dioxine-4-thiones 9 leads to the formation of acylthioketenes 10, which are characterized by Ar matrix IR spectroscopy as well as on-line tandem mass spectrometry. The thioketenes 10 undergo a 1,3-shift of the aryl group to generate thioacylketenes 11. Ketenes 11 cyclize to 3-aryl-thiet-2-ones 12, which are also characterized by matrix IR spectroscopy and tandem mass spectrometry. The thiet-2-ones 12 undergo two kinds of reaction under the FVT conditions: (i) cheletropic CO extrusion with formation of arylthioketenes 13, and (ii) cycloreversion to COS and arylacetylene. PMID- 10808444 TI - A highly enantioselective benzothiepine synthesis AB - A highly enantioselective synthesis of benzothiepine 1a has been accomplished via an enantioenriched sulfoxide intermediate obtained by asymmetric oxidation with a chiral oxaziridine in 89:11 er. The key step is a thermodynamically controlled asymmetric cyclization reaction that produces two new stereogenic centers. The (4R,5R) isomer 1a was obtained in 98:2 er. PMID- 10808445 TI - Tautomeric equilibrium of pyridoxine in water. Thermodynamic characterization by 13C and 15N nuclear magnetic resonance AB - A remarkable temperature dependence on the 13C NMR and 15N NMR chemical shifts of pyridoxine in water (pH = 7.0) has been observed. C-3, C-6, and N-1 were the most sensitive nuclei to the temperature effect. This dependence has been explained on the basis of an equilibrium shift thermally induced between the neutral and the dipolar form of this molecule. The thermodynamic characterization of tautomeric equilibria that interconvert quickly on the NMR time scale can be carried out from the observed average 13C NMR and 15N NMR chemical shifts at different temperatures (5-90 degrees C). We have developed a new method for the estimation of the thermodynamic parameters of a given equilibrium by fitting the experimental data to a theoretical curve. This new method allows us to improve the fitting results on our previously proposed methodology. We show that there are linear correlations between the average chemical shifts obtained from different nuclei at the same temperature. This indicates that the parameters of the pure forms are related among them. We have carried out a simultaneous multiple function curve fitting of all data obtained from the most sensitive signals together using these linear correlations as restricted conditions in order to diminish the number of independent parameters to fit. To test the new methodology, we have studied the thermodynamics of the tautomeric equilibrium of pyridoxine in water. We have obtained delta H degree values ranging from -23.6 +/ 1.3 to -25.8 +/- 1.7 kJ/mol for this equilibrium depending on the used data set. This kind of methodology has, among others, the following advantages: It allows the use of a great number of experimental points from different signals in the fitting process, it yields very precise and accurate values of the tautomeric process, and it allows the resolution of the problem with only 13C NMR data in some cases saving NMR time. PMID- 10808446 TI - EPR properties of two new cyclic phosphinylhydrazyl radicals and of their inclusion complexes with cyclodextrins AB - Two phosphorus-containing hydrazines, namely morpholin-4-ylphosphoramidic acid diethyl ester (1a) and (2,2,6,6-tetramethylpiperidin-1-yl)phosphoramidic acid diethyl ester (2a), have been synthesized. The corresponding hydrazyl radicals (1b and 2b) have been obtained, by in situ oxidation, and their properties have been investigated by EPR spectroscopy. The 1b radical shows spectra strongly dependent on temperature due to the inversion of the morpholin ring and to rotation about the N-N bond. Since, in the investigated temperature range, both motions take place in the EPR time scale, a kinetic study of these process could be made by analyzing the spectral line-shape variations. The 2b radical is highly persistent and shows a strong temperature and solvent dependence of the phosphorus splitting. The latter property was usefully exploited to study the guest-host interaction of this radical with cyclodextrins. A method is also proposed for the determination of affinity constants for cyclodextrins of nonparamagnetic compounds. PMID- 10808447 TI - Enantioselective photocyclization of amides to beta-lactam derivatives in inclusion crystals with an optically active host AB - Irradiation of inclusion crystals of 2-(N-acyl-N-alkylamino)cyclohex-2-enones and N,N-dimethylphenylglyoxylamide with chiral host molecules gave the optically active N-alkyl-1-azaspiro[3.5]-nonane-2,5-diones and 3-hydroxy-1-methyl-3 phenylazetidin-2-one, respectively. The crystal structure of the 1:1 inclusion complex of N,N-dimethylphenylglyoxylamide with (-)-trans-1,4-bis[3-(o chlorophenyl)-3-hydroxy-3-phenylprop-1-ynyl]-2,3,5,6- tetrachloro-2,5 cyclohexadiene-1,4-diol was analyzed by X-ray diffraction. PMID- 10808448 TI - Synthesis of novel push-pull unsymmetrically substituted alkynyl phthalocyanines AB - Two families of "push-pull" phthalocyanines 1-3 having an unusually strong dipole moment have been prepared. The syntheses of unsymmetrically substituted phthalocyanines 1a,b and 2 bearing one or two electron-withdrawing 4 nitrophenylethynyl moieties, respectively, and six alkoxy substituents were performed by combination of a zinc or nickel templated cyclotetramerization and cross-coupling palladium mediated methodologies. In a similar way, the "push pull" compounds 3a,b having a reversal substitution pattern, characterized by the presence of one electron-donor 4-(dimethylamino)phenylethynyl unit and six strong acceptor alkylsulfonyl substituents were prepared. The compounds show very large second-order nonlinear optical responses. PMID- 10808449 TI - Lipase-catalyzed transesterification of 2-hydroxy-2 (pentafluorophenyl)acetonitrile leading to (1R,2R)- and (1S,2S) bis(pentafluorophenyl)ethane-1,2-diol AB - Optically pure (1R,2R)- and (1S,2S)-1,2-bis(pentafluorophenyl)ethane-1,2-diol (1) were synthesized from key intermediates (R)- and (S)-2-hydroxy-2 (pentafluorophenyl)acetonitrile (2), both of which were prepared by the lipase LIP-catalyzed transesterification (E = 465). The absolute configuration of (S)-2 was determined by X-ray structural analysis after transformation into (S)-alpha cyano-2,3,4,5,6-pentafluorobenzyl (S)-6-methoxy-alpha-methyl-2-naphthaleneacetate (S,S)-9. In addition, the crystal structure of (S,S)-9 has an interesting well ordered packing pattern which shows face-to-face stacking interactions and end-to end parallel contacts between the pentafluorophenyl and 6-methoxynaphthyl groups of the adjacent molecules. PMID- 10808450 TI - Characterization, via ESR spectroscopy, of radical intermediates in the photooxidation of arylcarbinols by ceric ammonium nitrate AB - The photooxidation by ceric ammonium nitrate (CAN) of several aryl and naphthylcarbinols has been studied by means of ESR spectroscopy. For all the investigated arylcarbinols, but not for the naphthyl derivatives, it has been possible to detect radical intermediates deriving from the parent alkoxyl radicals. In particular, in the photooxidation of 1,1-diphenylethanol, a bridged radical intermediate has been detected. The assignment has been validated through experiments with two different labeled compounds: the 1,1-[2', 3', 4', 5', 6', 2", 3", 4", 5", 6"-2H10]diphenylethanol and the 1,1-diphenyl[2, 2, 2-2H3]ethanol. A similar bridged radical has been found to be formed in the photooxidation of triphenylmethanol, while, for the 1,1-diphenylpropanol, the only detectable species has been the ethyl radical deriving from a competitive beta-scission process. Finally, for the 2-phenylpropan-2-ol (cumyl alcohol), two radical species have been identified: the methyl, deriving from the beta-scission process, and the cyanomethylene, deriving from H-abstraction of the cumyloxyl radical from the solvent. A kinetic study on the competition of the two processes has also been conducted and the parameters of the Arrhenius equation for the latter process have been estimated. PMID- 10808451 TI - Preparation and NMR characterization of C70H10: cutting a fullerene pi-system in half AB - Three isomers of C70H10 were prepared by Zn(Cu) reduction of C70. Three chromatographic bands were identified as C70H10 species by MALDI-FT mass spectrometry, and these compounds were isolated by repeated HPLC treatments. The major isomer (2) was characterized by 1H and 13C NMR, while the minor isomers 3-4 were isolated in such small quantities that only 1H NMR analysis was possible. 1H coupled and 1H-decoupled 13C NMR of 2 established a 7,8,19,26,33,37,45,49,53,63 substitution pattern. This assignment was confirmed by HMBC and DFQ-COSY experiments. This structure is completely reasonable, as we found that 2 results exclusively from reduction of the 7,19,23,27,33,37,44,53-C70H8 that is formed in the course of the Zn(Cu) reduction of C70. PMID- 10808452 TI - ArF and KrF laser-induced gas-phase photolysis of selenophene and tellurophene: extrusion of Te and Se and intramolecular 1,3-H shift competing with beta-C-C cleavage in C4H4 residue AB - ArF (193 nm) and KrF (248 nm) laser-induced photolysis of gaseous selenophene and tellurophene (C4H4M, M=Se and Te) has been examined. It is shown that, unlike thiophene and furan, selenophene and tellurophene cleave both M-C bonds and yield the elemental heteroatom (Se, Te), 1-buten-3-yne, and ethyne. The proposed mechanism involves an intermediate .HC=CH-CH=CH. diradical that decomposes via two competitive pathways, namely, 1,3-H shift to 1-buten-3-yne and beta-cleavage to two molecules of ethyne. It is shown that the relative importance of the channels depends both on the energy of the photon and on the heteroatom. Specifically, the 1,3-H shift/beta-cleavage ratios are 2.3 (193 nm, M=Se), 3.6 (248 nm, M=Se), 1.4 (193 nm, M=Te), and 10.5 (248 nm, M=Te). The inertness of the Te residuum and the high preference for the 1,3-H shift in KrF laser photolysis of tellurophene suggest that this photolysis can serve as a source of the C4H4 diradical for mechanistic studies. PMID- 10808453 TI - Radical addition to isonitriles: a route to polyfunctionalized alkenes through a novel three-component radical cascade reaction AB - The reaction of aromatic disulfides, alkynes, and isonitriles under photolytic conditions affords polyfunctionalized alkenes--beta-arylthio-substituted acrylamides or acrylonitriles--in fair yields through a novel three-component radical cascade reaction. The procedure entails addition of a sulfanyl radical to the alkyne followed by attack of the resulting vinyl radical to the isonitrile. A fast reaction, e.g., scavenging by a nitro derivative or beta-fragmentation, is necessary in order to trap the final imidoyl radical, since addition of vinyl radicals to isonitriles seems to be a reversible process. The stereochemistry of the reaction is discussed, particularly with respect to the stereochemical outcome of related hydrogen abstraction reactions by the same vinyl radicals. The lower or even inverted preference for either geometrical isomer observed in our cases with respect to that encountered in hydrogen abstraction reactions is explained in terms of transition-state interactions and/or isomerization of the final imidoyl radical. The latter possibility is supported by semiempirical calculations, which show that the spin distribution in the imidoyl radical can allow rotation of the adjacent carbon-carbon double bond prior to beta fragmentation. PMID- 10808454 TI - Palladium-catalyzed cyclocarbonylation of o-iodoanilines with heterocumulenes: regioselective preparation of 4(3H)-quinazolinone derivatives AB - A catalyst system comprising palladium acetate-bidentate phosphine is effective for the cyclocarbonylation of o-iodoanilines with heterocumulenes at 70-100 degrees C for 12-24 h to give the corresponding 4(3H)-quinazolinone derivatives in good yields. Utilizing o-iodoaniline with isocyanates, carbodiimides, and ketenimines for the reaction, 2,4-(1H,3H)-quinazolinediones, 2-amino-4(3H) quinazolinones and 2-alkyl-4(3H)-quinazolinones were obtained, respectively. The nature of the substrates including the electrophilicity of the carbon center of the carbodiimide, and the stability of the ketenimine, influence the product yields of this reaction. Urea-type intermediates are believed to be generated first in situ from the reaction of o-iodoanilines with heterocumulenes, followed by palladium-catalyzed carbonylation and cyclization to yield the products. PMID- 10808455 TI - SET-induced photorearrangement of 2-phenylallyl phosphites. Stereochemistry at phosphorus. Application to cyclic nucleotide derivatives. AB - The stereochemistry at phosphorus of the SET-induced photorearrangement of diastereomeric 4-tert-butyl-2-phenylallyl-1,3,2-dioxaphosphorinanes (8) to the corresponding 2-phenylallylphosphonates (9), which involves exicted singlet 1,4 dicyanonaphthalene (1DCN*) as one-electron oxidant, was investigated. The rearrangement occurs with close to complete retention of configuration at phosphorus. The previously postulated mechanism for this photorearrangement is shown to be consistent with the stereochemical finding. Thus, one-electron reduction by DCN.- of the presumably stereospecifically formed distonic cyclic 1,3-cation radical intermediate 15, generated from cis-8 (Scheme 2), yields the thermodynamically stable diradical 16. beta scission of 16 forms phosphonate cis 9. An alternative mechanism involving beta scission of 15 to a styryl cation radical, prior to one-electron reduction to 15, is discounted on the basis of unpublished trapping studies using MeOH. The direct, kinetically controlled formation of diradical 16 rather than the thermodynamically less stable 21 with CH2 bonded apically to phosphorus is argued to be consistent with the essentially equal values of the quantum yield for phosphonate formation (phi P) on SET induced rearrangement of the acyclic 2-phenylallyl phosphite 1 and phosphite 7 with phosphorus incorporated in a six-membered (1,3,2-dioxaphosphorinane) ring. This mechanism is contrasted to that for the previously reported triplet sensitized photorearrangements of phosphites 1 and 7, which have greatly different phi P values. For these reactions, kinetic formation of the triplet analogue of 21, but without the tert-butyl substituent, requires a permutation of substituents for conversion to diradical 16 prior to intersystem crossing and beta scission to form the phosphonate corresponding to 7. The preparative-scale SET-induced photorearrangement of the thymidine-based 2-phenylallyl 3',5' phosphite 10 gave both diastereomers of phosphonate 11 that were separated by HPLC. The 2-phenylallyl functionality provides an opportunity for further functionalization. As reported elsewhere, 11 was not formed in useful amounts via triplet-sensitized reaction of 10. PMID- 10808456 TI - Triplet-sensitized photorearrangements of six-membered-ring 2-phenylallyl phosphites. Reaction efficiency and stereochemistry at phosphorus. PMID- 10808458 TI - A method for syn-dihydroxylation of double bonds cis to a hydroxymethyl substituent PMID- 10808457 TI - Novel ether-linked secondary face-to-face 2-2' and 3-3' beta-cyclodextrin dimers PMID- 10808460 TI - Synthesis of the marine natural product oroidin and its Z-isomer PMID- 10808459 TI - Nitroarylstannanes as synthons for the preparation of phenanthridine and benzo[i]phenanthridine derivatives. PMID- 10808461 TI - Diels-Alder reaction of heterocyclic imine dienophiles PMID- 10808462 TI - Claisen rearrangement of allyloxyanthraquinones with silver/potassium iodide in acetic acid as a new and efficient reagent PMID- 10808463 TI - Deoxygenative functionalization of hydroxy groups via xanthates with tetraphenyldisilane PMID- 10808464 TI - 2-oxohydrazones, 4-hydrazono-1H-pyrazol-5-ones, and derived products by air oxidation of 1,2-hydrazino-hydrazones PMID- 10808465 TI - First synthesis of nonracemic (R)-(+)-myrmicarin 217 PMID- 10808466 TI - High-yielding preparation of PMID- 10808467 TI - A simple, efficient, and general method for the conversion of alcohols into alkyl iodides by a CeCl3.7H2O/NaI system in acetonitrile PMID- 10808468 TI - Samarium(II) iodide promoted fragmentation and sequential reactions of aromatic 1,4-diketones PMID- 10808469 TI - Synthesis of 3,5-disubstituted 4-hydroxybenzoates by aryl-aryl and alkynyl-aryl coupling PMID- 10808470 TI - Substituted benzene anions as leaving groups in the reaction of sulfinyl derivatives with grignard reagents: a new and convenient route to dialkyl sulfoxides in high enantiomeric purity PMID- 10808472 TI - Analytic technique today. PMID- 10808471 TI - Aryl-fused nitrogen heterocycles by a tandem reduction--Michael addition reaction. PMID- 10808473 TI - The analyst's witnessing and otherness. AB - The analyst's active though silent witnessing of the patient's self-inquiry is presented as an essential aspect of the analytic process. Witnessing, though rooted in the analyst's empathy and holding, represents a more advanced development of those functions based on relational muturation from union to self other differentiation. Self-definition and regard for otherness are seen as intrinsically unitary. Psychoanalytic witnessing is first illustrated and defined, then located as a derivative of negation in the unfolding of the analytic process, next considered in relation to current concerns for intersubjectivity, and finally linked to current shifts in philosophical thought. PMID- 10808474 TI - Countertransference, conflictual listening, and the analytic object relationship. AB - Analytic listening is an ongoing conflictual process, containing all the components of conflict and shaped in every moment by both the patient's and the analyst's conflicts. The mutual responsiveness that develops between analyst and patient stems from a complex conflictual object relationship, fundamentally no different from any other object relationship, in which countertransference at all times simultaneously facilitates and interferes with the analytic work. Detailed clinical process is used to illustrate these and related phenomena, including the use of signal conflict, the benign negative countertransference, the function of countertransference structures, and the analyst's use of projection. The analyst's affects, thoughts, and actions trace the shifting nature of the patient's transference and resistance, and the level of the object relationship continuously being created between patient and analyst. PMID- 10808475 TI - Finding the neutral position: patient and analyst perspectives. AB - In light of current debates between classical and intersubjective schools of psychoanalysis, the challenge posed by the latter to such basic concepts as the analyst's neutrality, anonymity, and abstinence is taken up. It is maintained that the term neutral position is today more germane and meaningful than the term neutrality, which frequently has been taken to prescribe the analyst's posture. It is proposed that for each patient the neutral position is uniquely sited and that it is incumbent on the analyst to find its location. The neutral position is defined within the context of the interaction between analyst and patient. The concept is therefore compatible with--indeed it is essential to--an intersubjective or relational orientation. The manifold reasons, conscious and unconscious, why the analyst is vulnerable to leaving the neutral position are considered. The patient's reaction to the analyst who has left the neutral position and the analyst's clinical use of this reaction are discussed. PMID- 10808476 TI - The therapeutic and working alliances. AB - Since their introduction, the concepts of the therapeutic alliance and the working alliance have provoked debate regarding the nature and function of these alliances and the applicability and validity of the concepts. Features of these concepts as originally put forth by Zetzel and Greenson, respectively, are delineated, with emphasis on the significant distinctions between them. Their relation to degree of psychopathology is examined, especially with respect to what may be understood as the more "silent" aspects of the therapeutic alliance. Mutual identification, empathy, and role-responsiveness are stressed as constituent features of the therapeutic alliance, with the working alliance seen as possible (theoretically and clinically) only after a therapeutic alliance has to some degree been established. Both alliances are understood as intrinsic structures within the analytic process, and illustrative case material is presented. PMID- 10808477 TI - Love in the therapeutic alliance. AB - Volumes have been written about the patient's love for the therapist, but there has been relatively little discussion of the therapist's love for the patient. In an attempt to create a theoretical and technical space for discussing the appropriateness and role of love in the therapeutic relationship, a revised concept of the therapeutic alliance is applied to provide technical guidelines and understanding of two kinds of love between patient and therapist, corresponding to two systems of self-esteem regulation: an open, reality-oriented system and a closed, sadomasochistic system organized according to omnipotent beliefs. Examples of the role of love through the phases of treatment illustrate the interrelationship of love and the accomplishment of therapeutic alliance tasks. PMID- 10808478 TI - On the receiving end: facilitating the analysis of conflicted drive derivatives of aggression. AB - Freud's delay in giving aggression an instinctual role comparable to that of libido had clinical effects. Historically, this delay rescued psychoanalysis and its practitioners from destruction by Austria's anti-Semitic leadership. Since today the cultural threat to analysis is of quite a different nature, methods are proposed for making drive derivatives of aggression (and, in turn, libidinal drive derivatives) more accessible. Certain traditional methods have in significant degree distracted analysts from techniques that more effectively allow patients to bring conflicted derivatives of aggression into nonneurotic management through analytically facilitated ego growth. Approaches that move away from the concept of conflicted instinctual drives compound the basic resistance to analyzing defended drive derivatives of aggression. A recommendation is made for a two-tier choice of method: (1) a traditional technique for patients needing relatively less access to their aggression to achieve a satisfying outcome, and (2) close process attention for the analysis of analytic candidates and others for whom it is professionally advantageous to achieve greater insight into their aggressive potential. Recommendations are made regarding how the analyst can attain the neutrality necessary to conduct analyses of the latter sort without having to suffer narcissistic trauma while on the receiving end of verbally explicit, undisplaced drive derivatives of aggression. PMID- 10808479 TI - What is a deep interpretation? AB - Important differences are emerging regarding the place where analysts believe the most meaningful analytic work takes place. One area that highlights these distinct ways of working is the analyst's view of deep interpretations. Models underlying the differing perspectives on this issue are presented, along with an extended clinical example that illustrates the importance of considering, in formulating analytic interventions, the concept of a structured mind. A view of the analytic process that accords the patient's perspective greater privilege is introduced. PMID- 10808480 TI - Self-criticism and the psychic surface. AB - It is postulated that the surface of the mind reflects a variety of shifting self syntonic and self-dystonic contents striving to achieve a stable homeostatic balance. Shifts between self-syntonic and self-dystonic contents are seen to reflect an underlying intrasystemic conflict within the superego between conflicting superego injunctions. These shifts possess a cyclical quality, as self-syntonic contents become self-dystonic and as self-dystonic contents become self-systonic. This conceptualization of the psychic surface has implications for a comparative study of psychoanalytic technique and for implementing the technical recommendation to work from surface to depth. It is suggested that the analyst's interpretations become assimilated within cyclical processes of superego reaction and repair. PMID- 10808481 TI - [General and molecular ecology of Legionella]. AB - The review is devoted to the general and molecular ecology of bacteria of the genus Legionella in natural and anthropogenic environments. Invasion of amoebae and infusoria by legionellae and their replication in these protozoa can be considered to be a pre-adaptation for invasion of the human immune system. Symbiosis of bacteria and protozoa as a promising model of cellular microbiology and the conception of bacterial ecological niches are discussed in relation to the low fidelity of most bacterial species to their habitats (biotopes). The necessity of elaboration of a similar conception for microbial consortia and associations is emphasized. PMID- 10808482 TI - [Dark metabolism of acetate in Rhodospirillum rubrum cells, grown under photoheterotropic conditions]. AB - The mechanism of the dark assimilation of acetate in the photoheterotrophically grown nonsulfur bacterium Rhodospirillum rubrum was studied. Both in the light and in the dark, acetate assimilation in Rsp. rubrum cells, which lack the glyoxylate pathway, was accompanied by the excretion of glyoxylate into the growth medium. The assimilation of propionate was accompanied by the excretion of pyruvate. Acetate assimilation was found to be stimulated by bicarbonate, pyruvate, the C4-dicarboxylic acids of the Krebs cycle, and glyoxylate, but not by propionate. These data implied that the citramalate (CM) cycle in Rsp. rubrum cells grown aerobically in the dark can function as an anaplerotic pathway. This supposition was confirmed by respiration measurements. The respiration of cells oxidizing acetate depended on the presence of CO2 in the medium. The fact that the intermediates of the CM cycle (citramalate and mesaconate) markedly inhibited acetate assimilation but had almost no effect on cell respiration indicative that citramalate and mesaconate are intermediates of the acetate assimilation pathway. The inhibition of acetate assimilation and cell respiration by itaconate was due to its inhibitory effect on propionyl-CoA carboxylase, an enzyme of the CM cycle. The addition of 5 mM itaconate to extracts of Rsp. rubrum cells inhibited the activity of this enzyme by 85%. The data obtained suggest that the CM cycle continues to function in Rsp. rubrum cells that have been grown anaerobically in the light and then transferred to the dark and incubated aerobically. PMID- 10808483 TI - [Streptomyces sp. Z-11-6--a new producer of extracellular L-glutamate oxidase]. AB - Glutamate oxidase activity was studied in 1254 Streptomyces strains isolated from the zonal soils of various regions of Russia and other countries. Seven strains proved to be producers of extracellular L-glutamate oxidase. The most active producer strain was identified, and the conditions of enzyme biosynthesis were optimized. A multistep-mutagenesis and selection procedure allowed a genetically stable strain, Streptomyces sp. Z-11-6, to be obtained, whose glutamate oxidase activity was 40 times higher than that of the original natural isolate. PMID- 10808484 TI - [Extracellular L-glutamate oxidase from Streptomyces sp. Z-11-6: preparation of it and its properties]. AB - Mutagenesis induced with nitrous acid and subsequent selection allowed a genetically stable mutant strain, Streptomyces sp. Z-11-6, to be obtained, whose L-glutamate oxidase activity was 40-fold higher than that of the original natural isolate and was as great as 1.6-1.8 units/ml of culture liquid. A procedure for the isolation and purification of the enzyme was developed; the biochemical properties of the enzyme were studied. Out of 20 amino acids tested (including D glutamate), the glutamate oxidase from Streptomyces sp. Z-11-6 was active only with L-glutamate. This allows the concentration of L-glutamate to be determined in the presence of other amino acids. Calcium chloride at a concentration of 0.1 0.5% promoted the secretion of the extracellular glutamate oxidase. PMID- 10808485 TI - [Effect of culture media components on accumulation of glutaminyl endopeptidase in culture liquid of Bacillus intermedius 3-19]. AB - The effect of nutrients and growth conditions on the accumulation of glutamyl endopeptidase in the culture liquid of Bacillus intermedius 3-19 was studied. Glucose and other readily metabolizable carbon sources were found to suppress the production of the enzyme, while inorganic phosphate and ammonium cations enhanced it. Protein substrates, such as casein, gelatin, and hemoglobin, did not affect enzyme production. Some bivalent cations (Ca2+, Mg2+, Co2+) increased the production of glutamyl endopeptidase, but others (Zn2+, Fe2+, Cu2+) acted in the opposite way. The rate of enzyme accumulation in the culture liquid increased as the growth rate of the bacterium decreased, so that the maximum enzyme activity was observed in the stationary growth phase. Based on the results of this investigation, an optimal medium for the maximum production of glutamyl endopeptidase by B. intermedius 3-19 was elaborated. PMID- 10808486 TI - [Mutual effect of invertase and acid phosphatase from the yeast Saccharomyces cerevisiae on their secretion into culture media]. AB - The hypothesis that various extracellular enzymes produced by the yeast Saccharomyces cerevisiae exert a mutual influence on their secretion into the culture medium was tested experimentally. The statistically processed results indicate that extracellular invertase affects the secretion of acid phosphatase, and acid phosphatase affects the secretion of invertase. In addition, the secretion of each of these enzymes was shown to be subject to autoregulation. PMID- 10808487 TI - [Hemagglutinating activity of Lentinus edodes (Berk.) Sing [Lentinula edodes (Berk.) Pegler]]. AB - The hemagglutinating (HA) activity of the submerged mycelium and the culture liquid (CL) of four strains of Lentinus edodes was studied. The HA activity of CLs proved to be much higher than that of mycelia. The carbohydrate specificity of fungal agglutinating factors was determined. HA activity was investigated as a function of the inoculum size, cultivation temperature, and culture age. The agglutinating activity of different morphogenetic structures of L. edodes F-249, including mycelium, brown mycelial mat (MM), primordia, and fruiting bodies, was studied, MM was found to possess the maximum HA activity, which can be explained by the possible involvement of agglutinins in the formation of MM, which is composed of glued hyphae. PMID- 10808488 TI - [Mycotoxins from the fungi Penicillium vulpinum (Cooke & Massee) Seifert & Samson]. AB - Mycotoxins produced by seven strains of Penicillium vulpinum (formerly Penicillium claviforme) isolated from different sources were studied. The strains were characterized by specific profiles of secondary metabolites and produced mycotoxins of different structural types. In addition to toxins already known for this fungal species (patulin, roquefortine, 3,12-dihydroroquefortine, oxalin, viridicatin, cyclopenin, and alpha-cyclopiazonic acid), the strains studied also produced indolyl-3-acetic acid, griseofulvin, meleagrin, and cyclopeptin. PMID- 10808489 TI - [Dependence of transformation of chlorophenols by Rhodococci on position and number of chlorine atoms in the aromatic ring]. AB - Study of the conversion of chlorophenols by Rhodococcus opacus 1G, R. rhodnii 135, R. rhodochrous 89, and R. opacus 1cp disclosed the dependence of the conversion rate and pathway on the number and position of chlorine atoms in the aromatic ring. The most active chlorophenol converter, strain R. opacus 1cp, grew on each of the three isomeric monochlorophenols and on 2,4-dichlorophenol; the rate of growth decreased from 4-chlorophenol to 3-chlorophenol and then to 2 chlorophenol. The parameters of growth on 2,4-dichlorophenol were the same as on 3-chlorophenol. None of the strains studied utilized trichlorophenols. A detailed study of the pathway of chlorophenol transformation showed that 3-chloro-, 4 chloro-, and 2,4-dichlorophenol were utilized by the strains via a modified ortho pathway. 2-Chlorophenol and 2,3-dichlorophenol were transformed by strains R. opacus 1cp and R. rhodochrous 89 via corresponding 3-chloro- and 3,4 dichloropyrocatechols, which were then hydroxylated with the formation of 4 chloropyrogallol and 4,5-dichloropyrogallol; this route had not previously been described in bacteria. Phenol hydroxylase of R. opacus 1G exhibited a previously undescribed catalytic pattern, catalyzing oxidative dehalogenation of 2,3,5 trichlorophenol with the formation of 3,5-dichloropyrocatechol but not hydroxylation of the nonsubstituted position 6. PMID- 10808490 TI - [Teichoic acids from the cell wall of Nocardiopsis prasina BKM Ac-1880T]. AB - The cell wall of Nocardiopsis prasina VKM Ac-1880T was found to contain two structurally different teichoic acids: unsubstituted 3,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate), substituted at position 2 by 10% with alpha-N acetylglucosamine and by 5% with O-acetyl groups. The structure of the polymers was studied by chemical analysis and NMR spectroscopy. The results obtained correlate well with 16S rRNA sequence data and confirm the species-specificity of teichoic acids in the genus Nocardiopsis. PMID- 10808491 TI - [Effect of composition of cellular lipids of formation of nonspecific antibiotic resistance of alkanotrophic rhodocoocci]. AB - The antibiotic resistance and lipid composition of rhodococci grown in rich organic media with gaseous or liquid n-alkanes were studied. Hydrocarbon-grown rhodococci exhibited an increased resistance to a wide range of antibiotics (aminoglycosides, linkosamides, macrolides, beta-lactams, and aromatic compounds). The enhanced antibiotic resistance of rhodococci grown on n-alkanes correlated with an increased content of total cell lipids (up to 14-28%) and saturated straight-chain fatty acids (C16:0, C18:0, C21:0) and was accompanied by the appearance of cardiolipin and phosphatidylglycerol in cells. These lipid compounds were supposed to promote the formation of nonspecific antibiotic resistance in rhodococci by decreasing the permeability of their cell envelope to antibiotics. PMID- 10808492 TI - [Topological changes in DNA as a reflection of adaptation of Escherichia coli to oxidative stress under glucose starvation and transition to growth]. AB - Changes in the topological state of DNA occur in a starving Escherichia coli culture under oxidative stress caused by the addition of hydrogen peroxide. The addition of a carbon and energy source to this culture results in a second stress reaction. This supports previous data indicating that different mechanisms are responsible for the cell defense against oxidative stress in exponential and starving E. coli cultures. Polyamine synthesis is involved in the cell adaptation to the stress. Putrescine binding to DNA and its dissociation seem to modulate the DNA topological state, which regulates the expression of the adaptive genes. An increase in the activity of the polyamine-synthesizing system in response to oxidative stress leads to a putrescine flux across the cytoplasmic membrane, due to which the antioxidant activity of putrescine protects the membrane phospholipids and contributes to the restoration of the cell energy-generating function. PMID- 10808493 TI - [A family of shuttle vectors for lactobacilli and other gram-positive bacteria based on the plasmid pLF1311 replicon]. AB - A set of broad-host-range single-replicon shuttle vectors for cloning nucleotide sequences in gram-positive bacteria (lactobacilli, enterococci, lactococci, bacilli, etc.) was created. The vectors are based on the cryptic plasmid pLF1311 from Lactobacillus fermentum VKM 1311 belonging to a family of the sigma-type pE194-like plasmids. The vectors can replicate in gram-positive bacteria and Escherichia coli. They are stable in many gram-positive bacteria, have small sizes, and allow the selection of recombinants on media with X-Gal. The vectors that contain the region of initiation of the conjugal transfer of plasmid RP4 belonging to the incompatibility group IncP alpha can be mobilized in a great number of bacteria using a helper plasmid from E. coli but not from gram-positive bacteria. PMID- 10808494 TI - [Conjugative transfer of pTd33-plasmid between strains of Agrobacterium]. AB - Supramembrane structures that connect conjugating agrobacterial cells were visualized for the first time by transmission electron microscopy. The primary contact of cells during conjugation was shown to occur through the formation of long pili containing no VirB1 protein. Pretreatment of agrobacterial cells with acetosyringone resulted in a six- to tenfold increase in the transfer frequency of the plasmid pTd33 at 19-25 degrees C and had almost no effect at 30 degrees C. The transfer of the plasmid pTd33 from A. tumefaciens strain GV3101 to plasmid free A. tumefaciens strain UBAPF-2 was 16 times decreased after the centrifugation of cells. The transfer efficiency of the plasmid pTd33 from A. tumefaciens strain LBA2525 (virB2::lacZ) to plasmid-free A. tumefaciens strain UBAPF-2 was one order of magnitude lower than the transfer from the wild-type A. tumefaciens strain GV3101. Treatment of donor cells with 0.01% SDS before mating decreased the transfer efficiency by a factor of 26. The role of pili in the establishment of contact between conjugating cells of agrobacteria is discussed. PMID- 10808495 TI - [Roseinatronobacter thiooxidans Gen. Nov., sp. Nov., a new alkaliphilic aerobic bacteriochlorophyll-alpha-containing bacteria from a soda lake]. AB - Several samples of microbial mat obtained from soda lakes of the Kunkurskaya steppe (Chita oblast) abundantly populated by purple bacteria were screened for the presence of heterotrophic alkaliphiles capable of oxidizing sulfur compounds to sulfate. This capacity was found in only one pigmented strain, ALG 1, isolated on medium with acetate and thiosulfate at pH 10. The strain was found to be a strictly aerobic and obligately heterotrophic alkaliphile. Growth on medium with acetate was possible within a narrow pH range from 8.5 to 10.4. The strain formed a reddish orange carotenoid and bacteriochlorophyll a. Pigments were synthesized only at high concentrations of nitrogen-containing organic compounds (peptone or yeast extract). The production of bacteriochlorophyll a was maximal under microaerobic conditions in darkness. Strain ALG 1 could oxidize sulfide, thiosulfate, sulfite, and elemental sulfur to sulfate. In heterotrophically growing culture (pH 10), thiosulfate was not oxidized until the late logarithmic phase. The sulfur-oxidizing activity was maximal at the most alkaline pH values. The notable increase in the efficiency of organic carbon utilization observed in the presence of thiosulfate suggested that the bacterium was a sulfur-oxidizing lithoheterotroph. The phylogenetic analysis of the 16S rRNA gene showed strain ALG 1 to be a member of the alpha-3 subgroup of proteobacteria and to constitute a distinct branch located between nonsulfur purple bacteria Rhodobacter and Rhodovulum. Based on the unique phenotypic properties and the results of phylogenetic analysis, the alkaliphilic isolate ALG 1 was assigned to a new genus and species Roseinatronobacter thiooxidans with the type strain DSZM-13087. PMID- 10808496 TI - [Comparison of methods of storing lactic acid bacteria]. AB - The study was undertaken to elucidate how different methods of storage (immersing in mineral oil, lyophilization, and subculturing) of lactic acid bacteria belonging to the genera Lactobacillus and Lactococcus affect their viability, antibiotic activity, and ability to accumulate organic acids. Storage of the lactic acid bacterium L. lactis subsp. lactis by immersion in mineral oil proved to be ineffective. Lyophilization allowed the survival of a sufficiently large number of cells, although their antibiotic activity somewhat decreased. The resuscitation of lyophilized bacteria by subculturing them in rich nutrient media, such as skim milk, led to the restoration of their physiological activity, including the effective antimicrobial spectrum. PMID- 10808497 TI - [Physico-chemical and microbiological characteristics of groundwater from observation boreholes of a deep radioactive liquid waste repository]. AB - A radioactive liquid waste repository was found to be a habitat of the rich microbial community with a high catabolic potential. Groundwater from a depth of 162-189 m contained aerobic saprotrophic and anaerobic fermentative, sulfate reducing, and denitrifying bacteria. Nitrate-reducing bacteria residing in this groundwater were isolated in pure cultures. Based on the results of their physiological studies, 16S rRNA sequencing, and phylogenetic analysis, the microorganisms isolated were ascribed to one phylogenetic branch, the gamma subclass of gram-negative bacteria. Among six isolates, four belonged to the genus Acinetobacter, whereas two others belonged to the genera Comamonas and Aeromonas. The data obtained indicate that the microflora of the repository can exert a certain effect on the chemical composition of the formation fluids and bearing rocks, as well as on the migration of radionuclides. PMID- 10808498 TI - [Physiological and phylogenetic diversity of thermophilic spore-forming hydrocarbon-oxidizing bacteria from oil fields]. AB - The distribution and population density of aerobic hydrocarbon-oxidizing bacteria in the high-temperature oil fields of Western Siberia, Kazakhstan, and China were studied. Seven strains of aerobic thermophilic spore-forming bacteria were isolated from the oil fields and studied by microbiological and molecular biological methods. Based on the 16S rRNA gene sequences, phenotypic characteristics, and the results of DNA-DNA hybridization, the taxonomic affiliation of the isolates was tentatively established. The strains were assigned to the first and fifth subgroups of the genus Bacillus on the phylogenetic branch of the gram-positive bacteria. Strains B and 421 were classified as B. licheniformis. Strains X and U, located between B. stearothermophilus and B. thermocatenulatus on the phylogenetic tree, and strains K, Sam, and 34, related but not identical to B. thermodenitrificans and B. thermoleovorans, undoubtedly represent two new species. Phylogenetically and metabolically related representatives of thermophilic bacilli were found to occur in geographically distant oil fields. PMID- 10808499 TI - [Initial stages of interaction of Azospirillum brasilense bacteria with wheat germ roots: adsorption, deformation of root hairs]. AB - The initial stages of colonization of wheat roots by cells of Azospirillum brasilense strains 75 and 80 isolated from soils of the Saratov oblast were studied. The adsorption of azospirilla on root hairs of soft spring wheats rapidly increased in the first hours of incubation, going then to a plateau phase. Within the first 15 h of incubation, exponential-phase cells were adsorbed more intensively than stationary-phase cells. Conversely, stationary-phase cells were adsorbed more intensively than exponential-phase cells, if the period of azospirilla incubation with the wheat roots was extended. As the time of incubation increased, the attachment of azospirilla to the wheat roots became stronger. The effect of cell attachment to root hairs was strain-dependent; the number of adsorbed cells of a given strain of azospirilla was greater in the case of host wheat cultivars. The deformation of wheat root hairs was affected by the polysaccharide-containing complexes isolated from the capsular material of azospirilla. The suggestion is made that common receptor systems are involved in the adsorption of azospirilla on roots and in root hair deformation. PMID- 10808501 TI - [Phasic changes in reproductive potential, intracellular polyamines and antilysozyme activity in Escherichia coli in periodic culture]. AB - Growth-phase associated changes in and relationships between the specific growth rate (mu) characterizing the reproductive capacity of the cells, the contents of intracellular biogenic polyamines (BPA), such as putrescine (P), cadaverine (C), and spermidine (S), and antilysozyme activity (ALA) were studied in 37 strains of Escherichia coli grown in batch culture on solid medium. A decrease in mu upon the transition of the culture to the stationary growth phase was accompanied by a decrease in the pool of free BPA, mainly P and C, and by the appearance of ALA. The interrelations between the parameters studied were described as a complex of direct and negative correlations; the combination of low initial P and C contents, reduced P/S and C/S ratios, and a high level of ALA was designated as a factor of slight inhibition of E. coli reproduction. It is argued that BPA and ALA are integrated in a system controlling both the metabolism and stability of peptidoglycan in E. coli. PMID- 10808502 TI - [Intracellular phosphorus pool in a culture of the cyanobacterium Spirulina platensis]. AB - An intracellular phosphorus pool in a monoculture of the cyanobacterium Spirulina platensis was assessed using radioactive and nonradioactive phosphorus. The derived dependence of specific growth rate on the intracellular content of mineral phosphorus can be presented in the form of the Droop equation. It was found that the stage of replenishment of the intracellular phosphorus pool may affect the phosphorus turnover estimation in aquatic environments from the results of short-term measurements of phosphorus uptake. PMID- 10808500 TI - [Population dynamics of oligosporous actinomycetes in Chernozem soil]. AB - Investigation of the dynamics of an oligosporous actinomycete population in chernozem soil in the course of succession induced by soil wetting allowed us to reveal the time intervals and conditions optimal for the isolation of particular oligosporous actinomycetes. Saccharopolysporas and microbisporas proved to be best isolated in the early and late stages of succession, whereas actinomycetes of the subgroup Actinomadura and saccharomonosporas could be best isolated in the early and intermediate stages of succession. PMID- 10808503 TI - [Dopamine-dependent inhibition of glycine release in the rat nucleus accumbens during feeding]. AB - Food intake decreased the glycine extracellular level in the rat n.accumbens. Tetrodotoxin prevented the decrease, whereas D,L-threo-beta-hydroxyaspartic acid exerted no effect. Raclopride (D2 dopamine receptor antagonist) increased the glycine extracellular level in food intake. The data obtained suggest that during feeding the glycine release in the n.accumbens is controlled by the D2 dopamine receptors. PMID- 10808504 TI - [Participation of the rat caudate nucleus in the immunostimulating effect of DAGO]. AB - The data obtained suggest that administration of the DAGO increased the number of plaque- and rosette-forming cells after immunisation with the sheep red blood cells in the sham-operated Wistar rats. Following destruction of the caudate nucleus, the DAGO administration prevented the immune activation. Bilateral destruction of the caudate nucleus resulted in a considerable inhibition of the immune response as compared with the control rats. Thereupon the caudate nucleus seems to be involved in realisation of the DAGO-induced immune activation. PMID- 10808505 TI - [Serotonin metabolism in the rat brain during water deprivation and hydration]. AB - Water deprivation (WD) decreased the serotonin (5-HT) level and significantly increased the 5-hydroxyindoleacetic acid (5-HIAA) in the rat midbrain and hypothalamus, the catabolic 5-HIAA/5-HT ratio increasing three-fold. Hydration (H) produced a moderate increase in the 5-HT and 5-HIAA levels in the hypothalamus with no changes in the 5-HIAA/5-HT ratio. Hydration exerted no significant effect upon the 5-HT level and metabolism in the midbrain. A two-fold increase of corticosterone concentration in water deprivation and its decrease in hydration were shown to occur in peripheral blood plasma. PMID- 10808506 TI - [Endothelium function in ischemia-reperfusion (a literature review)]. AB - Endothelium was shown to be one of the most hypoxia-tolerant mammalian cell types and to be injured in ischemia to a lesser extent than other cells. Alteration in the endothelial function during hypoxia and ischemia protects the organs against ischemic injury. Postischemic reperfusion contributes to endothelial dysfunction and initiates the reperfusion injury. Some data show ischemia/reperfusion to cause a systemic alteration of endothelial function in remote organs. PMID- 10808507 TI - [Opioid system and cardiac resistance to ischemic and reperfusion injuries]. AB - In vivo pre-treatment with the opioid receptor antagonist D,L-naloxone completely eliminated the reperfusion-induced creatine kinase (CK) leakage from the rat isolated perfused haert. The inactive isomer L-naloxone decreased the CK release by half. The (-antagonist ICI 174,864 and k-antagonist nor-binanltorphimine exerted a weaker protective effect. The (-antagonist DAMGO, the (2-agonist DSLET, the k1-agonist spiradolin, or the sigma-agonist (+)-SKF 10047, improved myocardial cell viability after ischemia/reperfusion. PMID- 10808508 TI - [Nitric oxide storage in rats of various genetic strains and its role in the antistressor effect of adaptation to hypoxia]. AB - Adaptation to hypobaric hypoxia induced a gradual increase in the NO production along with a progressive NO storage in vascular wall. Unadapted August rats were more resistant against stress-induced stomach ulceration than the Wistar rats. Following a 6-day adaptation rats of both strains revealed a protective antiulcerogenic effect. A long-term adaptation potentiated the stress damage of the stomach rather than protected against it. A higher basal NO production seems to provide a more efficient antistress defence in the August rats. An intense NO storage may create a relative NO shortage and thus predispose to stress-induced vasoconstriction and ulceration. PMID- 10808509 TI - [Effects of parathyroid hormone and parathyroid hypertensive factor on myocardial mechanical properties in normotensive rats]. AB - After a 10-minute incubation, both the parathyroid hypertensive factor (PHF) and parathyroid hormone (PTH) reduced myocardial contractility and accelerated myocardial relaxation. Effects of the PTH and PHF on myocardial inotropic characteristics were of opposite directions. PMID- 10808510 TI - [Quantitative characteristics of the systemic anaphylaxis model in Sprague-Dawley rats]. AB - Anaphylactic response intensity was quantitatively estimated by means of measuring mean arterial pressure (MAP) and heart rate (HR). Damage to intestinal mucosa was studied by means of morphometry. These indices grew in a dose dependent way along with the amount of administered egg ovalbumin (OVA). The MAP and HR measurements seem to be useful in a quantitative elucidation of allergic sensitivity in laboratory animals. PMID- 10808511 TI - [Secret excretion from the mouse mammary gland]. AB - Histological studies revealed that the mammary gland nipple have smooth muscle fibres along the nipple channel. These fibres infiltrate the connective tissue parallel to the skin. The ring muscles are not obvious. Delays in the milk excretion in mice may be due to specifics of allocation and functioning of the nipple smooth muscles. To obtain milk, a mechanical action upon the nipple and a synchronised release of oxitocin into the blood are necessary. PMID- 10808512 TI - [Dependence of constrictor and dilator humoral responses of arteries and veins in the small intestine on calcium ions entering vascular myocytes]. AB - Verapamyl administered to the intestine circulation in the dose evoking no shifts in the arterial and venous resistance or in the vascular capacity, decreased 2- or 3-fold both the constrictor and dilatory responses of the arterial vessels. Against the background of Verapamyl, noradrenaline and isoproterenol did not practically change either capacity or resistance in the veins. Responses of veins to angiotensin II increased 10-fold and more, whereas they were absent after administration of atropine into the intestine circulation. Constrictor and dilatory responses of veins need to a greater extent than the arteries penetration of external calcium ions to their myocytes. PMID- 10808513 TI - [Effects of the bee and various snake venoms on the rat thermoregulation]. AB - The most sensitive organ to any kind of venoms is the skin, whereas muscles response of milder, and the intestine response reveals no difference between effects of different venoms. The isobarin desympathetising effect combined with V.lebetina venom shows that the venom affects thermoregulation interacting with the sympathetic nervous system, and exerts a sympathomimetic effect. This action seems to be not adrenergic as there is no compensation of the catecholamine decrease due to isobarin administration. PMID- 10808515 TI - [What does 'The Higher Nervous Activity Physiology' study?]. PMID- 10808514 TI - [Neuropharmacological analysis of participation of the serotoninergic system in hyperalgesia in neurotized rats]. PMID- 10808516 TI - [Psychophysiology as a behavioral science]. PMID- 10808517 TI - [Physiology of the higher nervous activity and psychophysiology]. PMID- 10808518 TI - [Physiology of the higher nervous activity and psychophysiology]. PMID- 10808519 TI - [Differences and similarities between physiology of the higher nervous activity and psychophysiology]. PMID- 10808520 TI - [Relations between psychophysiology and the higher nervous activity physiology]. PMID- 10808521 TI - [Does a concept of the higher nervous activity correspond to the modern state of the behavioral science?]. PMID- 10808522 TI - [Immune system physiology]. AB - Processes of interaction of immunocytes of different origin and with different cytokins, underlie the immune system mechanisms. The immune system structure, lymphocytes variety and properties, molecular basis of antigen recognition, were described. Dynamic structure of peripheral part of the immune system, immunocytes migration and re-circulation role in integration of lymphoid organs, were demonstrated. The molecular and cellular basis of the immune reaction and immune memory, were discussed. PMID- 10808523 TI - [Immunoglobulin E (IgE) physiology]. PMID- 10808524 TI - [Interethnic differences in the genetic control of the human immune status]. AB - Most parameters of the immune status were found to depend on the HLA-antigens determined in the phenotypes of the human subjects under study. The findings suggest a HLA-transmitted genetic regulation of immune response. Various HLA markers related to certain parameters of the immune status, were determined. These markers seem to be unique and different for each examined population. PMID- 10808525 TI - [Consequences of interactions between thymic lymphoid and epithelial cells in vitro]. AB - A 24-hour co-cultivation of thymocytes and epithelial cells taken from human thymus results in mutual activation of epitheliocytes and thymocytes, as well as in apoptosis of thymocytes. The apoptosis can also be induced by a cultural supernatant of the thymic-epithelial cells, its level being lower, however, than in the co-culture. Thymocyte death and elimination develop faster in a co-culture with allogeneic thymic epithelial cells. PMID- 10808526 TI - [Interleukin-1 in realisation of stress-induced changes in functions of the immune system]. AB - Stress influences of different duration and intensity induce production of a lymphocyte-activating factor (LAF) by murine peritoneal macrophages, and enhancement of Interleukin 1 (IL-1) level in the murine blood, inducing no alterations in the thymocyte reaction to concomitant action of the IL-1 beta which correlates with changes in the value of humoral immune response. The data obtained are in agreement with differently aimed stress-induced alterations in the activity of the membrane neutral sphingomyelinase: the key enzyme of the sphingomyelin cascade, in the membrane P2 fraction of the brain cortex. The IL-1 seems to participate in physiological mechanisms of realisation of stress reactions on the levels of its production and biological action on target cells as well as of the sphingomyelin pathway of its signal transduction in nerve tissue. PMID- 10808527 TI - [The role of neutral sphingomyelinase in the interleukin-1beta signal transduction in cells of the mouse cerebral cortex]. AB - Involvement of the sphingomyelin cascade in Interleukin 1 beta (IL-1) signal transduction pathway in membrane fraction P2 of the murine brain cortex, was found. A key role of the membrane enzyme neutral sphingomyelinase (nSMase) in triggering the sphingomyelin pathway for IL-1 beta, was confirmed. The IL-1 beta was shown to activate in a dose-dependent manner nSMase in the P2 fraction of the brain cortex. Employment of both brain cortex membranes from the mice deficient in the type I IL-1 receptor and of IL-1 receptor antagonist made it possible to obtain evidence on the necessity of the IL-1 beta binding to the type I IL-1 receptor for the nSMase activation. It appears that the IL-1 beta effects on the CNS are realized via IL-1 receptor type I and activation of the nSMase as an initiating enzyme of the sphingomyelin cascade. PMID- 10808529 TI - [Study of tonic and evoked electrical activity in efferent fibers of the sympathetic nerve in white rats]. AB - In anaesthetised Wistar rats, electrical sympathetic activity and a somatosympathetic reflex in the cervical sympathetic trunk elicited by a single electrical shock to forelimb or hindlimb afferent nerves, were recorded. The spontaneous activity was shown to conform with the pulse and respiratory waves of arterial pressure. Somatosympathetic reflex consists of early and late discharges evoked by somatic myelinated afferent fibres stimulation, and C-response elicited by stimulation of unmyelinated afferent fibres in spinal nerves. PMID- 10808528 TI - [Effect of hypothyroidism on 5-HT1A-, 5-HT2A-receptors and serotonin transporter in the rat brain]. AB - Effects of thyroid hormone deficiency on 5-HT1A receptors, 5-HT2A receptors and serotonin transporter in the brain were studied in thyroidectomised Wistar rats receiving an iodine-free diet and receiving 15 micrograms/kg of thyroxine for 21 days. Binding of 3H-8-OH-DPAT to 5-HT1A receptors and 3H-cytalopram to serotonin transporter were unchanged in hypothyroid rats as compared to the control. 3H ketanserin binding to 5-HT2A receptors was significantly decreased in the frontal cortex in hypothyroid rats. The cortical 3H-ketanserin binding in thyroidectomised rats was normalised after thyroxine replacement. The data suggest that the decrease in the cortical 5-HT2A receptors is the main consequence of impairing effect of hypothyroidism on serotonin neurotransmission. PMID- 10808530 TI - [Effect of the medium composition on the resting membrane potential in the earthworm longitudinal somatic muscle fibers]. AB - Norepinephrine or increased extracellular K+ hyperpolarize the membrane of the earthworm somatic muscle fibre, whereas removal of Cl- from external solution or a hypotonic solution depolarize the membrane. The dependence of the membrane resting potential on the extracellular K+ is quite characteristic against the background of ouabain action. A preliminary membrane depolarisation by ouabain eliminates the above effects on the membrane resting potential. The data obtained suggest that the ouabain-sensitive active ion pump directly contributes to the membrane resting potential value. This hypothesis is discussed with respect to existence of active Cl- transport combined with Na+, K(+)-pump which presumably takes part in the intracellular osmotic pressure regulation in the earthworm somatic muscle. PMID- 10808531 TI - [Modulation of the intensity of the non-quantal transmitter release by nitric oxide (NO) at the neuromuscular junction]. AB - In the rat diaphragm muscle, nitric oxide (NO)--sodium nitroprusside (SNP) and S nitroso-N-acetylpenicillamine (SNAP), as well as substrate for the NO synthesis L arginine, decrease the level of hyperpolarization of the muscle fibre membrane after acetylcholine receptor blockade by the d-TC and irreversible acetylcholinesterase inhibition by armin (H-effect). Contrary to that, disruption of the NO synthesis in the muscle fibres by the NO-synthase inhibitor NG-nitrol-L arginine methyl ester (L-NAME) results in enhancement of the H-effect both in vitro and in vivo. Inactivated SNP and inactive forms of arginine and NAME did not affect the H-effect magnitude. Haemoglobin, effectively binding the NO molecules, abolishes the suppressing effects of the SNP, SNAP and L-arginine upon the H-effect. The findings suggest that the NO could be acting as a modulator of nonquantal transmitter release at the mammalian neuromuscular junction. PMID- 10808532 TI - [Changes in tissue monoamine oxidase activity and resistance to acute hypoxia in rats during stress]. AB - Male Wistar rats exposed to different stresses developed shifts in the brain and liver monoaminoxidase activity. In the so called "cognitive" stimulation, the activity was enhanced in the brain and reduced in liver. Mild stresses also enhanced the activity in the brain. Extreme stimulation (starch peritonitis) caused a significant diminishing of the activity in the brain. All the stress schedules accompanied by enhancement of the brain monoaminoxidase activity increased the rats' tolerance of acute hypoxic hypoxia. Negative correlations between the blood lactic acid contents and the brain monoaminoxidase activity were revealed in rats of both the control and the "cognitive" groups. The findings suggest a direct interrelationship between post-stress shifts of the brain monoaminoxidase activity and the hypoxia tolerance. PMID- 10808533 TI - [The role of protein synthesis in realisation of protective cardiac effects of thyroid hormones during immobilization stress in rats]. AB - A decrease in the protein synthesis by the transcription inhibitor rifampycin prevents myocardial contractile function, protective effect of thyroid hormones and their antioxidant effect under the restraint stress in rats. The findings suggest that the main component of thyroid hormones' protective effect on the heart is related to their specific action realised via the cell genetic apparatus. PMID- 10808534 TI - [Study of functional properties of microvessels of the rat mesentery]. PMID- 10808535 TI - [The determination of chromosome fragile sites in the peripheral blood lymphocytes of patients with colorectal cancer taking into account the cancer pathology predisposition in the familial anamnesis]. AB - Results of comparative study of spontaneous and 5-bromdeoxyuridine-induced fragility of peripheral blood lymphocytes chromosomes in 9 patients with colorectal adenocarcinoma were presented. It was shown the increase of average spontaneous level of chromosomal fragility in patients with tumor aggregation in family as well as without it to 4.5 +/- 1.0 and 5.3 +/- 1.1 per 100 tested cells, accordingly. The increase of average level of damaged chromosomes in spectrum of rare sites to 12.5 +/- 2.6 in the patients with tumor aggregation in pedigree comparing to the patients without oncopathology in family 8.0 +/- 1.7 was observed. The most number of rare fragile sites was observed in 1q21 site of the chromosome 1. Possible connection between fragile sites of chromosomes in normal cells and malignant processes in the patients with colorectal cancer is discussed. PMID- 10808536 TI - [A cytofluorimetric study of the stability of the DNA secondary structure in the epithelial cells of adenocarcinomas of the large intestine]. AB - Investigation was carried out on the epithelial cells obtained from 32 patients with benign and malignant tumors of large intestine aged from 42 to 80 years. Ratio of single-stranded and double-stranded DNAs in the epithelial cells of unchanged colorectal mucosa (12 patterns), adenomatous polyps (4) and adenocarcinomas (29) was studied using fluorimetric analysis. Increased instability of DNA secondary structure was revealed in the tumor cells comparing to the cells of unchanged colorectal epithelium. Relative accumulation of single stranded DNA reflects structural and functional changes in gene apparatus of cells under malignization. PMID- 10808537 TI - [The characteristics of protein p53 expression in the mucosal epithelium of the large intestine affected by polyps and cancer]. AB - The direct dependence between p53 expression in epithelial cells and the level of morphological malignant neoplasm was established as the result of investigation of expression of this biomolecular marker in 12 polyps and 35 colorectal cancer. It was shown that family cancer syndrome 1 type was observed in pedigrees of majority patients with p53 positive cancers. PMID- 10808538 TI - [Ultrastructural disorders in the muscle fiber of the masticatory muscles in mandibular fractures]. AB - Basic impairments of the muscular fibre in fractures of the lower jaw with different localization within terms from several hours to several weeks in persons with various clinical course were studied with help of electronic microscopy. Following treatment of the injury with a "quiet" course, in the tissues studied there was activation of reparative processes up to normalization of the submicroscopic structures. Despite the given treatment, the injury accompanied with predominance of destructive changes in the muscular tissue. In the muscles, a disturbed permeability of depolarization waves was revealed, they causing contraction and relaxation of the muscles. PMID- 10808539 TI - [The duration of the meiosis stages in fertilized ovocytes of the mussel Mytilus galloprovincialis Lam. at different temperatures]. AB - Duration of meiosis steps in fertilized oocytes of mussel Mytilus galloprovincialis at three levels of development temperature optimum: 8.5, 12 and 17 degrees C. Absolute duration of meiosis stages increases with decreasing of water temperature. Relative duration of analogous stages doesn't depend on temperature conditions. Duration of first meiotic division determined by the time between maximal occurrences of consequent stages, at different temperatures is in two times greater than duration of second division. PMID- 10808540 TI - [The hybridization of tetraploid wheat with the octoploid wheat-wild rye amphiploid Elytricum fertile and the chromosome behavior in meiotic MI]. AB - The hybridization and chromosome behaviour regularities at meiotic MI of reciprocal F1 hybrids of tetraploid wheats T. durum and T. turgidum with 56 chromosome incomplete wheat-Elymus amphiploid (IWEA) Elytricum fertile, were investigated. Variation of main indices of the hybridization has been revealed to be determined by variance of such factors as cross direction, genotype peculiarities of wheat strains and their interaction. Elymus sibiricus genome, contained in IWEA karyotype, carries several genes, that can suppress wheat diploidization system. Reduction of homologous chromosome synapsis is not determined by these genes, but is connected with influence of other Elymus sibiricus chromosome factors. PMID- 10808541 TI - [The HLA system antigens in delayed sexual development]. AB - Frequency distribution of HLA antigens of A and B loci was examined in 138 adolescent boys aged 14-18 with delayed sexual development (DSD) of stages I-III, residing in the north-eastern region of Ukraine. Increase of A28- and B40 antigens and haplotypes A1-B40, A28-B40, A10-B40 determination frequencies was established. There were revealed positive and negative correlation between some of HLA antigens and DSD. Relative and attributive risks of DSD formation were calculated. PMID- 10808542 TI - [The immunophenotypic and cytogenetic characteristics of the blast cells in subvariants of acute myeloid leukemia]. AB - The results of study of morpho-cytochemical peculiarities, antigenic profile and peripheric blood and/or bone marrow blast cell karyotype of 21 patients with acute myeloid leukemia are presented. Hemopoietic cell immunophenotyping was carried out with the use of cytofluorometer FACScan and chromosome cytogenetic analysis with the use of analyzer "Metascan". It has been shown that in the AML M1 blast cell plasmatic membrane carries pan-myeloid CD33 and CD13 antigens, the last having high density of expression, and the CD38 antigen, which is a myeloid cell-precursor marker. In these patients tetraploidy, being the testimony of karyotype change, has been ascertained. It has been found out that the AML M2 blasts, except pan-myeloid antigens, express the cell proliferation CD71 marker. Blast cell karyotype peculiarities typical for this leukemia sub-variant have been revealed. In patients with the AML M4 in 3 of 6 cases an anomalous karyotype has been found. It has been also shown that the CD14 antigen, and rather its percentage in blast total population, is the differential-diagnostic criterion for the AML M4 and M5. PMID- 10808543 TI - [The activation of bovine oocytes to parthenogenetic development by ethanol]. AB - Dependence of parthenogenetic development ability of in vitro matured cow oocytes from their "aging" had been studied. It had been showed the possibility of parthenogenetic development of physiologically matured cow oocytes after inhibition of the first meiotic division and treatment with ethanol at least to 8 16-cell stage embryos. PMID- 10808544 TI - [The structure and possible nature of nuclear and cytoplasmic inclusions in plant cells]. AB - Literature data on structure, nature and possible functions of virus-induced inclusions as well as inclusions in normal plant cells were analyzed. Inclusions of various origins were classified basing on the literature data. Data on different types of inclusions in plant cells were summarized. PMID- 10808545 TI - [Ultrastructural organization of hypodermis and formation of cuticle in pharate larva of Leptotrombidium orientale (Acariformes: Trombiculidae)]. AB - The ultrastructural organization of hypodermis and the process of cuticle deposition is described for the pharate larvae of a trombiculid mite, Leptotrombidium orientale, being under the egg-shell and prelarval covering. The thin single-layered hypodermis consists of flattened epithelial cells containing oval or stretched nuclei and smooth basal plasma membrane. The apical membrane forms short scarce microvilli participating in the cuticle deposition. First of all, upper layers of the epicuticle, such as cuticulin lamella, wax and cement layers, are formed above the microvilli with plasma membrane plaques. Cuticulin layer is seen smooth at the early steps of this process. Very soon, however, epicuticle starts to be curved and forms particular high and tightly packed ridges, whereas the surface of hypodermal cells remains flat. Then a thick layer of the protein epicuticle is deposited due to secretory activity of hypodermal cells. Nearly simultaneously the thick lamellar procuticle starts to form through the deposition of their microfibrils at the tips of microvilli of the apical plasma membrane. Procuticle, as such, remains flat, is situated beneath the epicuticular ridges and contains curved pore canals. Cup-like pores in the epicuticle provide augmentation of the protein epicuticle mass due to secretion of particular substances by cells and to their transportation through the pore canals towards these epicuticular pores. The very beginning of the larval cuticle formation apparently indicates the starting point of the larval stage in ontogenesis, even though it remains for some time enveloped by the prelarval covering or sometimes by the egg-shell. When all the processes of formation are over, hungry larvae with a fully formed cuticle are actively hatched from two splitted halves of prelarval covering. PMID- 10808546 TI - [Cytotoxic and mitogenic effect of antimicrobial peptides from neutrophils on cultured cells]. AB - Cytotoxic and mitogenic activities of human and rabbit defensines (HNP and NP-2, resp.) and pig antimicrobial peptides from leukocytes (PR-39, prophenin PF-2 and protegrin PG-2) were studied. The above peptides were added to serum-free cell culture medium of the target cell lines K562, L929 and Hep22a. Cytotoxicity was estimated within 1, 3, 6, 24 and 48 h of cell incubation with the tested peptides in concentrations 1, 10, 25 or 100 micrograms/ml. All the examined peptides exhibited a distinct time- and concentration-dependent cytotoxicity. Moreover, by contrast to pig peptides, defensines could induce proliferation in cell subpopulations from cell lines L929 amd Hep22a, or L929 (defensines HNP and NP-2, resp.), keeping resistance to their cytotoxic action. PMID- 10808547 TI - [Thermo- and radioinduced enzymatic relaxation of superhelical DNA from mud loach Misgurnus fossilis L. spermatozoa]. AB - Actions of environmental impacts on mud loach spermatozoa were studied using various model systems: a) temperature stress, b) X-ray irradiation in vivo only of the animal head (a condition to trigger stress reaction), c) X-ray irradiation in vivo only of the animal body (a condition to exclude a direct activation of principal stress-realizing organism systems), d) gamma-irradiation in vitro of the cell suspension. It has been demonstrated that the temperature stress or X ray irradiation of the mud loach head induced three lines of effects: 1) significant decrease in DNA superhelical density, 2) activity redistribution (functional activation) of DNase II between chromatin subfractions (with the increase of its association to chromatin), and 3) intracellular acidification up to pH value to satisfy the DNase II initiation. The obtained facts allow to suggest that, first, DNase II participates in the presented temperature- and radio-induced supercoiled DNA relaxation in spermatozoa, and, second, DNase II is involved in physiological (season elimination of spermatozoa that remained within male gonads after fertilization) or environmentally-induced DNA degradation. PMID- 10808548 TI - [Osmotic modification of thermal damage in Escherichia coli bacteria at various pH values of the media]. AB - A study was made of the influence of media with different osmotic pressure on cell survival and on optic density of supernatants from Escherichia coli B/r and E. coli Bs-1 cell suspensions heated under different pH values of media. Hyperthermia induced cell death accompanied with the loss of optically active (lambda = 260 nm) material. Both cell damage effects were increased in acid and alkaline conditions, compared to neutral condition of heating. Hypertonic media results in a decrease in thermic cell death and loss of cell substances. Under this condition, the protection influence of high osmotic pressure was seen to increase significantly in acid and alkaline conditions of heating, compared to neutral condition. It has been proposed that a higher thermal damage of microorganisms in acid and alkaline beating conditions and protection influence of hypertonic media, especially expressed in acid and alkaline medium, is caused to a great extent by the status of osmotic cell homeostasis. PMID- 10808549 TI - [Functional properties and intracellular localization of high molecular weight isoforms of ligh chain myosin kinase]. AB - The vertebrate genetic locus, coding for a Ca2+/calmodulin-dependent enzyme myosin light chain kinase (MLCK), the key regulator of smooth muscle contraction and cell motility, reveals a complex organization. Two MLCK isoforms are encoded by the MLCK genetic locus. Recently identified M(r) 210 kDa MLCK contains a sequence of smooth muscle/non-muscle M(r) 108 kDa MLCK and has an additional N terminal sequence (Watterson et al., 1995. FEBS Lett. 373 : 217). A gene for an independently expressed non-kinase product KRP (telokin) is located within the MLCK gene (Collinge et al., 1992. Mol. Cell. Biol. 12 : 2359). KRP binds to and regulates the structure of myosin filaments (Shirinsky et al., 1993. J. Biol. Chem. 268 : 16578). Here we compared biochemical properties of MLCK-210 and MLCK 108 and studied intracellular localization of MLCK-210. MLCK-210 was isolated from extract of chicken aorta by immunoprecipitation using specific antibody and biochemically analysed in vitro. MLCK-210 phosphorylated myosin regulatory light chain and heavy meromyosin. The Ca(2+)-dependence and specific activity of MLCK 210 were similar to that of MLCK-108 from turkey gizzard. Using sedimentation assay we demonstrated that MLCK-210 as well as MLCK-108 binds to both actin and myosin filaments. MLCK-210 was localized in smooth muscle cell layers of aortic wall and was found to co-localize with microfilaments in cultured aortic smooth muscle cells. PMID- 10808550 TI - Evolutionary origin of eukaryotic cells. AB - This article reviews literature on the transition from rudimentary prokaryotic life to eukaryotes. An overview of the differences between these organisms and theories of eukaryogenesis are reviewed. Various methods of investigating the transformation from prokaryotes to eukaryotes are elaborated, including the fossil, the molecular and living records, and examples are given. Lastly, the recent molecular studies and the impact on phylogenetic classification for the tree of life, based on molecular evolution, are discussed. PMID- 10808551 TI - Isn't a picture still worth a thousand words? AB - The transmission electron microscope is a valuable diagnostic and research tool that is presently under-appreciated. In the area of human immunodeficiency virus research alone, it has provided critical information about viral pathogenesis and opportunistic infections and malignancies. However, because it has not always been used with care, the literature contains misinterpretations, especially as to what is a virus and what is actually a cell organelle, e.g., lysosome and Golgi vesicles. It is important to review the subject periodically to maintain its quality. PMID- 10808552 TI - Ultrastructural pathology of the heart in patients with beta-thalassaemia major. AB - Patients with beta-thalassaemia major frequently suffer from hypersiderosis which leads to hemochromatosis of major organs such as the heart and liver. Little information exists about the ultrastructural pathology of the human heart in beta thalassaemia patients. Five Cypriot patients with elevated blood ferritin and intractable heart failure were investigated. Cardiac biopsies from these patients were studied by light and electron microscopy, as well as by X-ray microanalysis. Ultrastructural examination revealed the presence of disrupted myocytes showing loss of myofibers, dense nuclei, and a variable number of pleomorphic electron dense granules. These cytoplasmic granules or siderosomes consisted of iron containing particles as confirmed by X-ray microanalysis. It is likely that the ultrastructural changes observed in myocytes of patients with beta-thalassaemia are largely due to iron deposition. PMID- 10808553 TI - Primary fibrosarcoma and malignant fibrous histiocytoma of bone--a comparative ultrastructural study: evidence of a spectrum of fibroblastic differentiation. AB - As primary bone fibrosarcoma (FS) and malignant fibrous histiocytoma (MFH) have similar clinical, radiographic, or survival manifestations, ultrastructural and immunohistochemical studies were undertaken to determine the differentiation pathways of constituent malignant cells. Twelve cases of primary intraosseous FS and MFH were selected for this ultrastructural comparative study and were analyzed for fibroblastic or modified fibroblastic differentiation. There were 4 FS cases and 8 MFH cases, of which 5 were storiform-pleomorphic, 2 were giant cell, and 1 was myxoid type. All FS consisted of spindle fibroblasts with a prominent rough endoplasmic reticulum and Golgi apparatus, variable amounts of vimentin intermediate filaments, and extracellular collagen fibrils. The MFH were composed of a mixture of spindle and pleomorphic fibroblasts (8/8), histiofibroblasts (4/8), and myofibroblasts (3/8). Variable numbers of undifferentiated cells were found in both tumors. In conclusion, fibroblastic differentiation and collagen production was noted in all cases. The often pleomorphic histiofibroblasts present in some MFH cases most likely represent "modified fibroblasts," similar to myofibroblasts. These findings support the hypothesis that the fibroblast and its variants are the predominant cell types found in these tumors, suggesting that the diagnostic entity MFH should be classified as a pleomorphic fibrosarcoma. PMID- 10808554 TI - Tuberin expression in ganglioglioma. AB - The expression of tuberin in neurosurgically resected gangliogliomas was investigated. Neither neoplastic astrocytes nor abnormal, multipolar large neurons showed immunoreactivity for tuberin. The reduced immuno-histochemical staining for tuberin was consistent with the reduction observed in Western blot analysis. Eosinophilic granular bodies and Rosenthal fibers were strongly immunoreactive for tuberin. The accumulation of tuberin in astrocytes with intracellular degenerative changes suggests increased expression in reactive cells, and perhaps a broader role for tuberin in central nervous system disease. PMID- 10808555 TI - Detection of VP-16-treated HL-60 cell apoptosis by TUNEL electron microscopy. AB - Apoptosis is essential to many physiological processes, including maturation of cells and the immune system. Deficient regulation of apoptosis may play an important role in many pathological conditions, such as autoimmunity, AIDS, and myelodysplastic syndrome. Several methods have been described to identify apoptotic cells. DNA strand breaks can be identified by labeling free 3'-OH termini with modified nucleotides by enzymatic reaction (TUNEL method). In this study, apoptosis was introduced in cultured HL-60 cells treated with VP-16, and the TUNEL method was adapted for electromicroscopy, called the EM TUNEL method. The results of the EM TUNEL method were compared with those of light microscopy and flow cytometry. Apoptotic cells following VP-16 (10 micrograms/mL) treatment were detected after 3 h by all methods (light microscopy, Erythrocin B, electron microscopy, flow cytometry, TUNEL, and EM TUNEL). EM TUNEL was the most sensitive method of detection, with a detection rate of 32%. Furthermore, EM TUNEL was more suitable for distinguishing cell lineage than light microscopy TUNEL. The results indicate that EM TUNEL can be used to detect apoptosis in bone marrow species in vivo. PMID- 10808556 TI - Electron microscopy in pathology articles: a retrospective appraisal. AB - Electron microscopy (EM) is a valuable standard tool in basic research and teaching. However, its use in diagnosis is limited, either for strategic reasons or budgetary constraints. This means that its many potential applications are more often neglected, either as an ancillary tool, quality control method, or gold standard, to complement, support, or confirm results of pathological studies. To evaluate the use of EM in this setting, the authors analyzed all articles (n = 2,531) in the three top indexed diagnostic pathology journals for a period of 60 months from July 1993 to June 1998. A total of 448 articles in which the use of EM was indicated, according to standard surgical pathology textbooks, were selected. Both the actual and the potential EM content of each article were scored, as follows: zero, illustrative, supportive, gold standard (for confirmation of research results), extensive, and predominant. Of the total number of articles in which EM was indicated, 77% made use of the technique. EM support was lacking most frequently in articles on serosal neoplasms and on new diagnostic strategies (p < .00005). There was no definite trend toward an increase or decrease in the use of EM during the period analyzed. The authors conclude that EM is used in most reports on diagnostic pathology, when it is indicated. However, a small but non-negligible percentage of articles (23%) could benefit from including EM as an ancillary, control, or gold standard method to complement, support, or confirm their results. PMID- 10808557 TI - Pigmented carcinoma of the breast: an ultrastructural study. AB - A pigmented skin lesion on a breast removed for carcinoma resembled melanoma by routine light microscopy, but correlation with immunohistochemistry and electron microscopy established that carcinoma cells within the upper dermis were intermingled with a proliferation of non-neoplastic melanocytic cells. Ultrastructurally, the tumor cells possessed desmosomes and intracytoplasmic lumina and mature melanosomes were present in their cytoplasm. The melanocytic cells were identified as melanocytes or melanophages, and it was concluded that the tumor in the skin was a passively pigmented carcinoma and not a melanoma or metaplastic breast carcinoma. PMID- 10808558 TI - Sinonasal teratocarcinosarcoma: ultrastructural and immunohistochemical evidence of neuroectodermal origin. AB - The authors report a case of sinonasal teratocarcinosarcoma in a 37-year-old man, which was located in the anterior skull base and extended to the right nasal cavity and paranasal sinuses. The tumor was surgically resected twice, but it could not be removed completely. Microscopically, it was mainly composed of primitive cell nests within a moderately cellular stroma. The components of squamous cell epithelia with focal teratoid appearance and adenocarcinomatous differentiation were observed. There were many rhabdomyoblasts scattered in the nests and stroma. Ultrastructurally, the primitive cells had many neural processes with parallel microtubules, resembling olfactory neuroblastoma. Rhabdomyoblasts showed various degrees of skeletal muscle differentiation. Some of the stromal spindle cells had actin filaments with dense patches and dense core granules. Immunohistochemically, the primitive cells were positive for epithelial markers, neuron-specific enolase, synaptophysin, and myogenic regulatory proteins. The rhabdomyoblasts showed immunoreactivity for myoid markers, cytokeratin, epithelial membrane antigen, and synaptophysin. Most of the stromal spindle cells were positive for smooth muscle actin, neuron-specific enolase and synaptophysin. The immunohistochemical and ultrastructural findings suggest that primitive cells had the most primitive phenotype of placodes, and support the possibility that sinonasal teratocarcinosarcoma is essentially a neuroectodermal tumor with divergent differentiation. PMID- 10808559 TI - Human endogenous retroviral expression by leukocytes. AB - Budding endogenous retroviral particles were identified on the plasma membrane of a high percentage of neurotrophils in four widely varied pathological specimens. PMID- 10808560 TI - Comment to: gastric carcinomas with unusual cytoplasmic ultrastructural features. PMID- 10808561 TI - Annulate lamellae in gastric carcinoma. Re: Gastric carcinomas with unusual cytoplasmic ultrastructural features. Caruso RA. Ultrastruct Pathol. 1999;23:275 276. PMID- 10808562 TI - [An improvement in the demonstration of the causative agents of dangerous infections under the conditions of local wars and armed conflicts]. AB - The system of the specific indication of the causative agents of infectious diseases, accepted in the Armed Forces of the Russian Federation (RF) in piece and wartime and based on the study of specimens by stages in laboratories of the sanitary epidemiological institutions of the Ministry of Defense of the RF, is presented. This system is insufficiently adapted to the conditions of local wars (LW) and armed conflicts (AC). Under these conditions, it is expedient to carry out the specific indication of infective agents not by stages, but completely (according to an enlarged scheme), replenishing the medical service of the field forces with specialists, transferred from higher sanitary epidemiological institutions. Specialized formations of the sanitary epidemiological institutions of the Center should be deployed neat the region of LW or AC, or the specimens should be promptly brought to the Center for indication by air. The improvement of the system of indication includes the introduction of new indication methods, such as polymerase chain reaction, into practical work and cooperation with the institutions and formations of the Ministry of Health of the RF. PMID- 10808563 TI - [The use of molecular biological methods for the individual characterization of Mycobacterium tuberculosis strains]. AB - The expediency of using molecular biological methods for the evaluation of M. tuberculosis clinical strains by individual genetic certification of circulating M. tuberculosis strains has been substantiated. Considerable genetic heterogeneity of M. tuberculosis strains isolated from patients in different regions of the Russian Federation has been established; this heterogeneity is due to the presence of differences in the number of copies (5-26) of element IS6110 in M. tuberculosis cells. PMID- 10808564 TI - [Noncultivatable forms of Francisella tularensis]. AB - Conditions for the appearance of F. tularensis uncultivated forms and for their reversion into the initial state have been studied. As revealed in this study, the combined influence of stress factors (starvation and low temperature) may result in the transition of F. tularensis into the uncultivated state in which it persists in the environment during the period between epidemics. The reversion of F. tularensis uncultivated forms into the initial state has been carried out with the use of sensitive animals. The uncultivated state of F. tularensis should be regarded as the actual form of the existence of the causative agent of tularemia in soil and water ecosystems. PMID- 10808565 TI - [The multiplication dynamics of pathogenic bacteria in relation to the trophic and temperature cultivation conditions]. AB - The comparative study of the dynamics of multiplication of Listeria monocytogenes and Yersinia pseudotuberculosis in organic and synthetic media and in distilled water at temperatures of 37 degrees C and 6 degrees C was carried out. This study revealed that in organic media the multiplication of bacteria was good at 37 degrees C and 6 degrees C. In mineral media and distilled water their multiplication was observed only at 6 degrees C. Moreover, conditions necessary for the multiplication of pathogenic bacteria in distilled water were shown; these conditions depended on the inoculated dose, the number of autolyzed microbial cells and the state of the culture. Proofs of the multiplication of the bacteria under the conditions of minimum nutrition and low temperature were obtained with the use of labeled 3H-thymidine. PMID- 10808566 TI - [The cultivation of Vibrio cholerae with green algae in an experiment]. AB - Relationships between Vibrio cholerae of different origin and some serogroups with green algae Scenedesmus quadricauda in mineral medium at two temperatures have been experimentally studied. Differences in the relationships of various strains with green algae under the above-mentioned experimental conditions have been established. The study has shown that a decrease in the concentration and the death of vct+ and vct- vibrios of all strains under study occur in the linear phase of the development of algae. 3 V. cholerae strains, serogroups O139 (vct+) and O50 (vct-), have been shown to be capable of survival under the conditions mix cultivation with algae for 50-100 days. The perish of green algae is supposed to increase the survival time and multiplication of V. cholerae under experimental conditions. PMID- 10808567 TI - [A comparison of the yield of Vi and O antigens during the adaptation of Salmonella typhi strains to cultivation under starvation conditions]. AB - S. typhi strains Ty(2)4446 and Vi-1S underwent multiple passages in f synthetic liquid starvation culture medium consisting of water with salts and glucose added. In the process of the adaptation of the cultures to these stress conditions (starvation stress) the increasing yield of biomass from passage to passage was observed. Differences in the accumulation of Vi- and O-antigens were noted in two strains under study. In the cultures of strain Ty(2)4446 an insignificant increase in the antigen content from passage to passage was observed, while in the cultures of strain Vi-1S an increase in the content of Vi- and O-antigens was 4- to 5-fold. With the adaptation of the culture the Vi antigen to O-antigen ratio changed from 1:57 to 1:20 for strain Ty(2)4446 and from 1:2.7 tp 1:2.2 for strain Vi-1S. Strain Ty(2)4446 had an advantage over strain Vi-1S with respect to the synthesis of Vi-antigen. These data are indicative of the expediency of using not only strain Ty(2)4446, but also strain Vi-1S for the preparation of typhoid vaccine, especially the one based on Vi antigen. PMID- 10808568 TI - [An evaluation of a nutrient medium for isolating and cultivating Helicobacter pylori]. AB - Selective Helicobacter agar containing the selective supplement and blood, adding ex tempore, for the isolation and cultivation of H. pylori was developed. The Helicobacter agar was studied with the use of 5 newly isolated H. pylori strains, 13 bacterial associated cultures, as well as 21 inoculated biopsy specimens of the gastric and duodenal mucosa of patients with peptic ulcer. The study revealed that Helicobacter agar ensured the growth of H. pylori and their isolation from clinical material. The positive results after the inoculation of the specimens of biopsy material on Helicobacter agar and control media was 85%. In addition, the study of Helicobacter agar showed that it also exhibited pronounced selective properties with respect to bacterial associations, not inhibiting the growth of Helicobacter organisms and retaining their main biological properties. It is possible to recommend Helicobacter agar for use in laboratory practice in diagnosing Helicobacter-associated diseases. PMID- 10808569 TI - [A dried nutrient medium for the isolation of Vibrio cholerae]. AB - A dried differential nutrient medium for the isolation of V. cholerae has been developed. The medium is sufficiently sensitive, has pronounced differentiating properties and greatly inhibits the appearance of microbial associations. During the cultivation of V. cholerae with the use of this medium the cultural, morphological and agglutination properties of the initial strains are retained. PMID- 10808570 TI - [The laboratory diagnosis of an outbreak of hemorrhagic fever at Oblivskaya village, Rostov Province: proof of the etiological role of the Crimean-Congo hemorrhagic fever virus]. AB - The results of the molecular biological detection of the etiologic agent of hemorrhagic fever in Rostov Province are presented. The role of the causative agents of Astrakhan rickettsial fever, hemorrhagic fever with the renal syndrome, Q fever, leptospirosis and listeriosis has been excluded by means of such immunochemical reactions as the direct and indirect immunofluorescent tests, the solid-phase immunoenzyme assay, the complement fixation test and the agglutination test. The relationship between the cases of hemorrhagic fever in the focus of the outbreak and Crimean-Congo hemorrhagic fever virus has been demonstrated due to the use of the polymerase chain reaction with preliminary reverse transcription. PMID- 10808571 TI - [Crimean-Congo hemorrhagic fever in Rostov Province: the epidemiological characteristics of an outbreak]. AB - The results of the epidemiological analysis of the outbreak of hemorrhagic fever which was caused by Crimean-Congo hemorrhagic fever virus and occurred during the period of July 3-19, 1999, in the Oblivskaya district of Rostov Province are presented. The specific epidemiological features of the outbreak have been determined. The possible versions of the appearance of the focus of infection and the role of Ixodes ticks in the circulation of the infective agent are discussed. PMID- 10808572 TI - [The reactogenicity and safety of a multicomponent vaccine (VP-4) in the prevention of acute respiratory diseases in preschoolers]. AB - The reactogenicity and safety of poly-component vaccine (VP-4), prepared from the antigens of opportunistic bacteria, in the prophylaxis of acute respiratory diseases (ARD) in children aged 2.6-6 years. The vaccine was administered intranasally in 3 administrations and orally in 6-8 administrations at intervals of 3-4 days for a period of 24 +/- 4 days. The prophylaxis of ARD with the use of VP-4 was carried out in 168 children in 4 children's preschool institutions. The control group was made up of 120 children, attending the same institutions. The study revealed that VP-4 had low reactogenicity and induced short-time systemic and local reactions (common cold, cough). The administration of VP-4 at a period of the epidemic rise on influenza and ARD morbidity did not lead to an increase in the frequency and duration of ARD in the vaccinees, as well as to the exacerbation of chronic infection and the allergization of the body. PMID- 10808573 TI - [The vaccinal prophylaxis of hepatitis B in Russia--the achievements, problems and outlook]. AB - Certain specific features of the present epidemic situation with hepatitis B (HB) in Russia were established: significant growth of HB morbidity, starting from 1995; the prevalence of persons aged 15-29 years among HB patients, which was linked with the sharp activation of the sexual route of the transmission of HB virus in recent years; an essential increase in the number of patients having contacted this virus in the process of the intravenous use of drugs. The results of the use of vaccine "Engerix B" among persons belonging to different risk groups were considered (a decrease in HB morbidity among them by 8-19 times was noted), the study demonstrated high immunogenicity anti-HBs antibodies on protective titers were determined in 92.3-95.7% of the vaccinees) and low reactogenicity of the vaccine, as well as stable postvaccinal immunity (5 years after the course of vaccination was completed anti-HBs antibodies were retained in 70.6-74% of the vaccinees). The study showed that only the vaccination of adolescents in combination, in the presence of opportunity, with the immunization of newborn infants and young children in the first year of their life made it possible to produce an essential effect on the activity of the epidemic process. Already in 2 years such organization of work on the prophylaxis of HB in one of the cities of the Sverdlovsk region led to a decrease in HB morbidity by 2.9 times, and among adolescents 9 times. PMID- 10808574 TI - [The evaluation of the reactogenicity, harmlessness and prophylactic efficacy of Grippol trivalent polymer-subunit influenza vaccine administered to schoolchildren]. AB - Vaccine "Grippol"--has been developed at the State Research Center--Institute of Immunology. The preparation belongs to new generation vaccines and is a trivalent polymer-subunit vaccine containing the sterile conjugate of influenza virus surface proteins, types A and B, bound with copolymer polyoxidonium. The administration of "Grippol" to children of school age (6-18 years) demonstrated low reactogenicity of the vaccine, its safety and sufficient prophylactic effectiveness. During observations on total morbidity (with the exception of influenza and acute respiratory diseases) no side effects produced by "Grippol" were registered. At the same time the fact that the morbidity rate of upper respiratory tract disease in the group of children immunized with the vaccine decreased in comparison with the control group (by 2.4 times) cannot be disregarded. PMID- 10808575 TI - [A comparison of the immune response induced by DNA or by an inactivated vaccine against tick-borne encephalitis]. AB - BALB/c mice were immunized with recombinant plasmid DNA pSVK3-ENS1 and pcDNAI-NS3 containing, respectively, genes E-NS1 and NS3 of tick-borne encephalitis (TBE) virus. Antibodies to TBE virus proteins were detected in the blood sera of the immunized animals by the method of the enzyme immunoassay. Though the titers of virus-specific antibodies in the sera of mice immunized with protein vaccines exceeded those registered after immunization with DNA vaccines, essential protective immunity was observed after the use of both vaccines. PMID- 10808576 TI - [The immunity characteristics of men with gastric and duodenal peptic ulcer associated with Helicobacter pylori]. AB - In males with peptic ulcer of the stomach and the duodenum, associated with H. pylori, the population and subpopulation spectrum of peripheral blood lymphocytes was studied by the method of CD typing with the use of monoclonal antibodies manufactured by the Research Institute "Preparat" (Nizhny Novgorod, Russia). The study revealed the development of disturbances in the amount of populations and subpopulations of peripheral blood lymphocytes, accompanied by a decrease in the absolute number of T lymphocytes (CD3, CD7), T helpers (CD4), T cytotoxic lymphocytes (CD8), B lymphocytes (CD22) and the ratio of CD4/CD8 cells. A decrease in the amount of IgA, IgG, the key complement components C1, C3, C5 was established. In peptic ulcer the immunological characteristics of gastric juice were found to essentially differ from those of saliva by a lower content of mucin, IgA and a higher content of SIgA and IgM in gastric secretions, which may form prerequisites for the colonization of the gastric mucosa by H. pylori. PMID- 10808577 TI - [An evaluation of the effect of 2,4-dichlorophenoxyacetic acid derivatives on humoral and cellular immunities]. AB - A single oral administration of phenoxy herbicides, viz. 2,4 dichlorophenoxyacetic acid (2,4-D), amino salt, sodium salt, diethylene glycol, iso-octyl and octyl ethers in a dose of LD16, was found to produce a pronounced suppressive effect on humoral immunity on day 5. The 2,4-D derivatives under study, administered in the same dose, stimulated the manifestations of delayed type hypersensitivity in mice immunized with sheep red blood cells. The effect of the inhibition of antibody formation to thymus-dependent antigen ranged in the order of the increase in the toxicity of substances: amino salt, diethylene glycol ether, 2,4-D, sodium salt, iso-octyl and octyl ethers. On the contrary, the intensity of delayed-type hypersensitivity was more pronounced with regard to substances having greater toxicity. PMID- 10808578 TI - [The antibacterial resistance of the strains making up the components of probiotic preparations]. AB - The spectrum of the antibiotic resistance of 21 strains of normal microflora, mainly forming constituents of widely used probiotics to 25 antibacterial preparations. Lactobacillus spp. were prevalent in the strains under test. The spectrum of the antibiotic resistance of lactobacilli varied, the gradation of resistance being more pronounced with respect to strains and not species. Both highly sensitive and highly resistant Lactobacillus strains were found: L. acidophilus (a component of biopreparation "Linex"), L. plantarum 8RA3, L. fermentum 90T4C (probiotic "Lactobacterin"), L. fermentum BL96. Bacteria used as the components of combined preparation "Linex" exhibited the highest resistance to a number of modern antibiotics. Strains of bifidobacteria were found to be highly sensitive to antibiotics. PMID- 10808579 TI - [Cloning and gene expression in lactic acid bacteria]. AB - The possibility of using the genera Lactobacillus and Lactococcus as vector representatives is widely discussed at present. The prospects of the construction of recombinant bacteria are closely connected with the solution of a number of problems: the level of the transcription of cloned genes, the effectiveness of the translation of heterologous mRNA, the stability of protein with respect to bacterial intracellular proteases, the method by protein molecules leave the cell (by secretion or as the result of lysis). To prevent segregation instability, the construction of vector molecules on the basis of stable cryptic plasmids found in wild strains of lactic acid bacteria was proposed. High copying plasmids with low molecular weight were detected in L. plantarum and L. pentosus strains. Several plasmids with molecular weights of 1.7, 1.8 and 2.3 kb were isolated from bacterial cells to be used as the basis for the construction of vector molecules. Genes of chloramphenicol- and erythromycin-resistance from Staphylococcus aureus plasmids were used as marker genes ensuring cell transformation. The vector plasmids thus constructed exhibited high transformation activity in the electroporation of different strains, including L. casei, L. plantarum, L. acidophilus, L. fermentum and L. brevis which could be classified with the replicons of a wide circle of hosts. But the use of these plasmids was limited due to the risk of the uncontrolled dissemination of recombinant plasmids. L. acidophilus were also found to have strictly specific plasmids with good prospects of being used as the basis for the creation of vectors, incapable of dissemination. In addition to the search of strain-specific plasmids, incapable of uncontrolled gene transmission, the use of chromosome-integrated heterologous genes is recommended in cloning to ensure the maximum safety. PMID- 10808580 TI - [The classification of streptococci and human streptococcal diseases]. AB - The modern nomenclature, phenotypic, medical, ecological and phylogenetic classification of streptococci and different classification of streptococcal human diseases are presented. All phylogenetic groups of streptococci have been shown to contain species causing diseases in man. The most medically significant groups are the phylogenetic groups Pyogenes and Mitis. Directions of the improvement of the classification of streptococci and streptococcal human diseases on the basis of modern concepts on the taxonomy of streptococci, the biological properties and ecology of the infective agents, as well as the genesis and clinical picture of diseases induced by them, have been determined. PMID- 10808581 TI - [The prevention of infectious diseases in light of the law on The Sanitary and Epidemiological Well-Being of the Population]. PMID- 10808582 TI - [The moving forces of the epidemic process]. PMID- 10808583 TI - [Local anti-infective protection of the reproductive tract in women of different ages]. AB - Healthy girls and women of the reproductive age, as well as women immediately before and after menopause, were examined. Neutrophils and immunoglobulins of cervical and vaginal secretions were studied and, as a result, age-dependent differences in the activity of the anti-infectious protection of the reproductive tract of women were found. PMID- 10808584 TI - [The characteristics of local immunity in Chlamydia-associated chronic inflammatory diseases of the organs of the lesser pelvis in women]. AB - Local immune reactions in 107 women with Chlamydia-associated chronic endometritis and salpingo-oophoritis were studied on endometrial biopsy specimens. Of these women, group 1 consisted 45 patients with Chlamydia associated chronic endometritis and salpingo-oophoritis and group 2 consisted of 62 such patients having, in addition, bacterial vaginosis and candidiasis. Among associated bacteria, epidermal staphylococci, Escherichia coli and their combinations prevailed. Pathomorphologically, in the biopsy specimens taken from the patients of group 1 a decrease in the activity of mononuclear phagocytes, secondary SIgA deficiency with the level of IgA-producing plasmocytes relatively unchanged, the moderate intensity of the synthesis of IgG and a shift in the ratio of T-helpers/T-suppressors towards the latter were observed. In the biopsy specimens of the uterine mucosa obtained from group 2 a decrease in the phagocytic activity of neutrophil granulocytes and macrophages, the pronounced suppression of IgA production by IgA plasmocytes with a sharp decrease in the amount of SIgA and an essential increase in the number of IgG-producing cells were noted. Disturbances of local immunity, observed in the presence of associated Chlamydia infection, gives grounds for the inclusion of immunomodulating preparations into the complex of therapeutic measures. PMID- 10808585 TI - [The local immunity of the genital system in pregnant women with a genital infection]. AB - In 101 healthy pregnant women and 132 pregnant women with genital infection (colpitis, endocervicitis, cervicitis) the characteristics of the anti-infectious protection of cervical mucus were studied. In pregnant women with inflammatory diseases of the vagina and cervix uteri disturbances in the local immunity of the sex system were detected. The study showed that in genital infections local immunodeficiency depended to a greater extent on the localization and spread of the process rather than on the etiological factor of the disease. PMID- 10808586 TI - [The role of different microorganisms in the genesis of tubal-peritoneal infertility and in immunity]. AB - The character of microflora, directly causing inflammatory process in tuboperitoneal infertility, was studied. The study revealed the prevalence of microbial associations, Chlamydia infection was detected in half of the women, and in 17% of the patients this infection was for the first time detected in the upper section of their reproductive apparatus, which was indicative of the role of Chlamydia infection in the occlusion of the uterine tubes. Opportunistic microflora persisting in persons with defective antimicrobial protection was found. The study of local protective factors in peritoneal fluid revealed the absence of T-cell activity, which was manifested by a low level of sIL-2R and the absence of a significant rise in the level of IL-2 in peritoneal fluid in comparison with that in healthy women. In addition, a decrease in the total complement activity and in the activity of component C3 of the complement was established. PMID- 10808587 TI - [The microflora of the reproductive tract in women during intrauterine interventions]. AB - The species compositions and persistence factors of the vaginal and cervical microflora of the reproductive tract of women in cases of intrauterine interventions (medical abortion, intrauterine contraception) were studied. Women with inflammatory complications following intrauterine interventions were found to have the same species of bacteria in their vaginal and cervical microflora. In addition, an increase in the values of the persistence factors of vaginal microflora was registered in women practicing intrauterine contraception and a decrease in the persistence potential of vaginal microflora was registered after abortion. PMID- 10808588 TI - [The correction of dysbiosis of the female reproductive tract by using an estradiol-containing preparation]. AB - The influence of Ovestin, an estradiol-containing preparation, on the vaginal microflora in women of the reproductive age in dysbiosis was studied. As the result of the local application of this preparation, the normalization of microflora was noted in 92% of women, which was manifested by an increase in the number of lactobacteria and indigenous corynebacteria, a decrease in the number, or elimination, of opportunistic microorganisms and the suppression of their capacity for persistence. PMID- 10808589 TI - [Mixed urogenital infection in men]. AB - High occurrence of sexually transmitted diseases (STD) caused by a combination of different infective agents was established. In 62.5% of cases such diseases took a course typical for mixed infections. Combinations of infective agents of STD were found to have several typical associations, e.g. Ureaplasma urealyticum and Mycoplasma hominis in combination with opportunistic microflora. The microbiocenosis of the reproductive tract was shown to form under the influence of a complex of endogenous (natural biological) and exogenous (sexual behavioral) factors leading, in each particular case, to the formation of stable associations. PMID- 10808590 TI - [The role of the microbial factor in the pathogenesis of male infertility]. AB - Microorganisms can be isolated from most seminal fluid samples, but the significance of bacteriospermia is uncertain because many males lack symptoms associated with the bacterial infection of the reproductive tract. The data on the influence of urogenital tract infections of fertility are contradictory. In many cases opportunistic microorganisms cause such classical infections of the urogenital tract as epididymitis and prostatitis, as well as subclinical reproductive tract infections. Some possible pathophysiological mechanisms of the development of infertility linked with infection of the ejaculate are considered: its direct effect on the fertile properties of the seminal fluid due to a decrease in the number of spermatozoa, the suppression of their motility, changes in their morphology and fertilizing capacity, its indirect influence due to the inhibition of spermatogenesis resulting from testicular damage, autoimmune processes induced by inflammation, secretory dysfunction of the male accessory sex glands as a consequence of the infection of the reproductive tract organs, leukocytospermia with its secondary influence of the ejaculate parameters, etc. The importance of the pathogenic properties of microorganisms for the localization of the inflammatory process in the urogenital tract of males and their role in the pathogenesis of male infertility are discussed. PMID- 10808591 TI - [Nonenteric escherichiosis and the problem of human reproductive health]. AB - The contribution of nonenteric Escherichia infections (NEI) to the formation of male and female infertility is analyzed. The negative influence of NEI, and particularly pyelonephritis, on the course of pregnancy, parturition and the postnatal period is shown. The role of E. coli in the development of life threatening meningoencephalitides, pneumonia, sepsis in newborn infants is emphasized, NEI risk factors are systematized with due regard to concrete stages in the pathogenesis of this pathology. Phenotypic differences between the causative agents of NEI and nonpathogenic and diarrhea-inducing E. coli are characterized. PMID- 10808592 TI - [The laboratory diagnosis of cytomegalovirus infection in infants in the first months of life]. AB - The detection rate of IgM, specific to cytomegalovirus (CMV), in the umbilical blood was 2.0 +/- 1.8% in infants with antenatal complications in their medical history, 8.9 +/- 1.8% in hospitalized infants aged 7 days to 6 months; simultaneously, such IgM proved to be absent in healthy infants of the same age. The maximum primary detection rate of CMV-specific IgM in hospitalized infants was registered at the age of 1.5-4 months and the minimum detection rate, at the age of 6 months, as well as at the age of 1 month and younger. In sick infants aged 0-6 months CMV-specific IgM were detected, as a rule, for 2-4 weeks. 86% of infants with CMV-specific IgM detected in their blood were found to have cytomegaloviremia; in 80% of patients the virus was excreted with saliva and in 50% the virus was detected in blood plasma. PMID- 10808593 TI - [The role of staphylococci in the occurrence, development and chronicity of lactation mastitis]. AB - The investigation results obtained by the authors and the data of literature on the role of the biological properties of staphylococci in the pathogenesis, development and chronization of acute suppurative lactation mastitis are generalized. The concept of lactation mastitis as a monoetiological disease caused solely by bacteria of the genus Staphylococcus is substantiated. The importance of the individual properties of staphylococci at different stages of the pathogenesis of lactation mastitis, as well as their role in ensuring the resistance of the infective agent to the protective mechanisms of the body and in the chronization of the suppurative process, is considered. The data on the use of a number of biological properties of staphylococci as markers for the prognostication of the course of lactation mastitis are presented and their prognosticating effect at different tactics of surgical treatment is evaluated. PMID- 10808594 TI - Accelerated bone mineral loss in HIV-positive patients. PMID- 10808595 TI - Psychiatry and HIV: Part 2. PMID- 10808596 TI - Perinatal guidelines for use of antiretroviral drugs updated. PMID- 10808597 TI - Minorities with AIDS getting less appropriate care than whites. PMID- 10808598 TI - Ethical guidelines on HIV vaccine research. PMID- 10808599 TI - $16 million in substance abuse treatment grants available. PMID- 10808600 TI - Glaser Pediatric Research Network Launched. PMID- 10808601 TI - Spanish HIV/AIDS Q&A forum. PMID- 10808602 TI - Yale and University of Connecticut receive $6 million to fight the spread of HIV. PMID- 10808603 TI - Bristol-Myers Squibb launches $2 million AIDS grant program. PMID- 10808604 TI - Is there a role for immunotherapy in controlling HIV infection? AB - Although HAART restores immune function in patients with HIV infection, restoration is incomplete. Functional restoration is seen primarily in responses to antigens that are prevalent in HIV-infected persons. Immunization is required to restore responses to antigens that are not predictably present. As an exception, HIV-specific responses are also generally not restored despite the prevalence of these antigens. This may be because HIV replication specifically targets and either destroys or renders nonfunctional HIV-reactive CD4+ T cells. Perhaps because HIV selectively targets HIV-reactive immune cells, therapeutic immunization strategies are particularly important areas of investigation in the treatment of HIV disease. Strategies designed to restore HIV-specific CD4+ T-cell function must also enhance the activity of HIV-specific cytolytic T cells, since these are the likely key mediators of defense against HIV replication. Both active immunization strategies and treatment interruption strategies may enhance HIV-specific immune responses. Treatment interruption, by increasing exposure to HIV antigens through heightened HIV replication, also runs the risk of permitting sufficient HIV replication to damage HIV-responsive CD4+ cells as well as enhancing the losses of other CD4+ cell populations that may protect against opportunistic complications of HIV disease. Thus, treatment interruption strategies require careful and sophisticated monitoring and should not be tried at home. PMID- 10808605 TI - Can antiretroviral therapy ever be stopped? AB - A pool of latently infected CD4+ T cells is established within the first few weeks of HIV infection. Because these memory T cells are inactive, the viral DNA integrated into their chromosomes remains invisible to immune surveillance and to antiretrovirals. Research shows that this reservoir of infected memory T cells does not decay in a clinically meaningful time frame--that is, within 60 years- in patients being treated with potent combination antiretrovirals. Even a hypothetical regimen that prevents any new infection of cells would not hasten the decay of this latent reservoir. Treatments that activate this reservoir have been studied in patients with suppressed viremia, but such interventions are highly toxic and have not succeeded so far. Studying rare individuals who manage to control activation of these latent cells may provide important clues to long term control of HIV infection. PMID- 10808606 TI - Clinicians' discussion: trends in HIV care. PMID- 10808607 TI - Preventing mother-to-child transmission of HIV: practical strategies for developing countries. AB - The HIV pandemic has greatly affected women of childbearing age in developing countries and, thus, their offspring, through mother-to-child transmission (MTCT) of the virus. Scientific advances, most of them established by randomized clinical trials, have recently led to the development of practical strategies aiming to reduce the public health burden of MTCT of HIV. These advances came first in non-breast-feeding populations, for example, in Thailand and, more recently, in African populations, where breast-feeding remains the predominant mode of infant feeding. This article reviews major accomplishments in this area, outlines practical issues for program implementation, and suggests future research needs. Short-course zidovudine and short-course nevirapine in the peripartum period currently represent 2 valid options to reduce MTCT of HIV in developing countries if appropriate prenatal, obstetrical, and postnatal care is provided and if alternatives to breast-feeding are considered according to the local situation and the mother's individual decision. PMID- 10808609 TI - 2nd European Graduate Student Meeting of the German Pharmaceutical Society. Frankfurt, Germany, 3-5 March 2000. Abstracts. PMID- 10808608 TI - HIV prevention in the era of HAART: implications for providers. AB - This article reviews the medical literature regarding the association between reduced HIV viremia caused by use of HAART and the risk of HIV transmission, finding that there is a correlation but that it is not absolute. It then examines the behavioral literature, which indicates that there is evidence, though far from conclusive, that, for some, increased sexual risk taking may be related to attitudes and beliefs about combination therapy. The article also considers the message about HIV prevention being presented by public health programs and ways in which health care providers can outline changing circumstances and enable the patient to make informed choices in "gray areas." PMID- 10808610 TI - [Evaluation of the survival time of complete dentures with a mathematical method]. AB - Based on the number of previous dentures and on the information given by the patients the authors describes a mathematical model for the estimation of the denture survival time. The distribution of edentulousness according to age (based on 9991 patients) shows a regular pattern and can be described by a second degree function. The survival time shows a similar regularity. The application of the Kaplan-Meier method can be used to estimate the survival time calculated only from a single cross-sectional study. This approach uses the survival time of the first set of complete dentures and the average edentulous time of the patients with their second complete dentures as a base for the calculation. PMID- 10808611 TI - [Prevalence of the various types of periapical lesions and the significance of histologic evaluation]. AB - To determine the prevalence of radicular cyst and chronic apical periodontitis among human periapical lesions 299 lesions were analysed during the past 6 years. The specimen were obtained during extraction or endodontic surgery. Of the lesions assessed, 195 (65.2%) were found histopathologically to be cystic, 96 (32.1%) were diagnosed as chronic apical periodontitis and 8 (2.7%) were observed as "other lesion". Studies to determine the diagnostic features and prevalence of these lesions have failed to reach a consensus view, though the treatment and the prognosis may differ to the lesion present. PMID- 10808612 TI - [Blood flow measurements in human oral tissues with laser Doppler flowmetry]. AB - In our examination the non-invasive laser Doppler flowmetry (LDF) was used to determine the blood flow in the tooth pulp (PBF) and in the gingiva (GBF) in humans. Perfusion values were collected by the means of LDF probe adjusted manually or by manipulator assisted fixation. Significantly lower GBF rate was detected by hand-held probe as compared to that obtained by manipulator assisted fixation (p < 0.001). No significant difference was observed for the PBF rates (p > 0.9). The coefficient of variation determined for GBF was higher (0.32 +/- 0.09) in case of manual procedure than at manipulator assisted (0.13 +/- 0.02) fixation mode. In contrary, the value of variation coefficient of PBF data obtained by hand-held probe was lower as compared to that calculated from data of manipulator fixation (0.16 +/- 0.08 v. 0.22 +/- 0.1; p < 0.05). These data indicate that while the manual fixation for measuring PBF seems to be satisfactory, for the accurate determination of GBF rate application of manipulator is essential. PMID- 10808613 TI - [Expression of oncogenes (c-myc-neu) and prolactin receptor (PRLr) in tissues of women with endometriosis]. AB - Endometriosis is a disease of unknown ethology; there are several theories that are unproven. It is known that women with such disease present with infertility, and that recurrence is very high, and apparently not depending of estrogenic stimulus. This phenomenon has been contributed to other growth factors and some oncogens. There are few studies about oncogens or other hormonal receptors; so, this study tries to evaluate them in eutopic tissue and in ectopic one of women with previous diagnosis of endometriosis. Our results show that myc as PRLr are expressed differently in the different tissues; and neu is expressed in the same way in both tissues. So, it was concluded that the cells of the endometriosis focus, have a differential status relating to expression of some of their genes, which target on their development and maintenance in a hormonal environment which differs from the uterine cavity. PMID- 10808614 TI - [DDT and related pesticides in maternal milk and other tissues of healthy women at term pregnancy]. AB - Ten healthy patients with term pregnancy resolved by abdominal via. During the surgical procedure samples of umbilical chord serum and maternal adipose tissue, were taken. In all the samples, together with the maternal milk collected the day 10 of puerperium, the concentrations of the following organochlorine pesticides, were measured up: (PCC); beta-BHC; gamma-BHC; heptachloride; aldrin; dieldrin, DDE, DDD, DDT and methoxychloride. The identification and quantification of pesticides was done by the comparison with standards certified by NIST (National Institute of Standard Technology). The general characteristics of the participants were: primigestas of 24.1 years aged, married and of a low socioeconomical level. As to the neonates, of 39 weeks of gestational age, female sex; 3,311 g of corporal weight and size of 51.1 cm. In all the analyzed samples at least one of the organochlorine pesticides was present. The results of correlation analysis between DDT concentration, present in the maternal serum with those identified in the adipose tissue and serum from the umbilical chord were highly significant: a = 0.97 and 0.87, respectively. In the maternal milk the highest concentrations of total DDT, were found, average of 2053 ng/g lipidic base, which is 2.8 times more of daily accepted intake. Likewise, DDT concentration in maternal serum kept exponential relation, growing with age (a = 0.99). PMID- 10808615 TI - [Tumor necrosis factor-alpha and interleukin-1 beta in maternal, fetal and retroplacental intravascular compartments at term and preterm labor]. AB - Neonato preterm birth (before 37 pregnancy weeks) account more than 80% perinatal deaths not attributable to congenital malformations. Preterm and term labor full mechanisms are unknown at present. Proinflammatory cytokinesis direct participation have been involved in the phenomena by several experimental evidence. The study's aim was to determine TNF-alpha and IL-1 beta concentration at maternal, fetal and fetal-maternal vascular compartments in women with term and preterm delivery and in women at term childbirth without labor. TNF-alpha and IL-1 beta concentration were determinated by commercial immunoassay. TNF-alpha concentration showed a tendency to be in more proportion at fetal and fetal maternal compartments in preterm and term childbirth groups versus TNF-alpha concentration in term group without labor at same places. IL-1 beta concentration showed same tendency of increase than TNF-alpha in preterm and term childbirth groups, but alone at fetal-maternal compartment. Statistical difference were not documented at any compartment or group compared. Data allow to identify fetal maternal compartments as target places where TNF-alpha and IL-1 beta were synthesized. Gradient concentration synthesis of cytokinesis allows to intend fetus as TNF-alpha initial producer. PMID- 10808616 TI - [Comparative evaluation of bone mineral density in Mexican women using x-ray bone densitometry and ultrasound]. AB - With lifestyle changes in women, smoking and use of beverages with caffeine, and sedentarism increasing, so the risk factors for decalcification, increase; which is a public health problem by the higher incidence of osteoporotic fractures, as the age advances, specially in the postmenopause woman, which means a greater secondary morbidity-mortality; an important cause of physical disability, which directly affects psychoemotional wellbeing in women. In this study two methods of bone densitometry, were used; one of x ray, and other with ultrasound in 138 women during postmenopause with an average index of corporal mass of 29. Both results were compared of bone density, T measurement with osteopenia and osteoporosis. Double densitometry, was done in the 138 patients of lumbar spine with DEXA equipment, and of calcaneum with DTU-one equipment, by the same technician, finding the difference of T punctuation in this double study. PMID- 10808617 TI - [Apoptosis in trophoblast of patients with recurrent spontaneous abortion of unidentified cause]. AB - Recurrent spontaneous abortion of unidentified cause or idiopatic is an important problem of reproduction health. In this study, the study has been started of one of the mechanisms that could act in the pathology of human pregnancy. The general hypothesis proposed, is that in the case of patients with recurrent spontaneous abortion of non identified cause (RSA) there are phenomenons in the materno/fetal/placentary means that manifest themselves in harm to the functionality and/or vitality of placentary tissue. The hypothesis was consistent with the finding of massive activation of cellular death in all the cases with RSA studied in this work. PMID- 10808618 TI - [Culture of blastocysts using sequential media]. AB - The advantages of cultivating human pre-embryos to the stage of blastocites, are well known. The use of media of culture sequential, without the backing of somatic cells, is new. The objective was to cultivate human pre-embryos to the stage of blastocyte to the determine the recuperation indexes, implantation index and pregnancy index in patients subjected to FIVT/TE or ICSI. Once obtained the ovules of patients were injected/inseminated to latter be cultivated for 72 hours using P1 medium at 10% of SSS under mineral oil for, latter, be transferred to a complex medium for blastocites culture for 48 hours, and finally be transferred. Thirteen were included (9 of FIV and 4 of ICSI) obtaining 205 ovules; fertilized 143 and 131 had cellular division. One hundred and twenty one pre-embryos were cultivated to blastocyst stage, from which, 53 reached that stage (43.8%); transferring 28 and freezing 25. In average, 2.1 blastocysts were transferred by patient. There were four pregnancies and one alive newborn, at term for an index of implantation of 14.2% and a pregnancy index of 30.7%. The study shows our initial experience, which demonstrated and acceptable idea of recuperation of blastocysts and pregnancies. PMID- 10808619 TI - 11th World Congress of the International Society for the Study of Hypertension in Pregnancy. Kobe, Japan, October 26-30, 1998. Abstracts. PMID- 10808620 TI - The continual challenge of empowering children and families. PMID- 10808621 TI - Medicaid managed care and children with special health care needs: a case study analysis of demonstration waivers in three states. AB - Although the number of states moving toward managed care for Medicaid beneficiaries is increasing, little is known about the extent to which managed care delivery systems are prepared to address the special needs of children with chronic illness and disabilities. This study explores the change in the process of care provided by the State Title V Programs for Children With Special Health Care Needs (CSHCN) and in parent and provider satisfaction when Medicaid changes the method of payment from fee-for-service to capitation. The findings suggest that CSHCN are vulnerable in a managed care environment and that states should proceed cautiously to integrate this population into their managed care arrangements. PMID- 10808622 TI - Explaining infant feeding style of low-income black women. AB - Unstructured interviews were conducted with 10 low-income black women to explore infant feeding style. Formula-feeding with early introduction of cereal in the bottle was the most common pattern used by mothers in the first 3 months. By 6 months, formula-fed infants had a complex diet of a variety of foods. Half the women intended to breast-feed, but only one exclusively breast-fed. Beliefs about healthy infants and crying influenced feeding. There was a lack of knowledge about and support for breast-feeding in these women's environment. Support and advice about infant feeding from the health care system were uneven. PMID- 10808623 TI - Parenting an infant during alcohol recovery. AB - This qualitative study explored women's concurrent experiences of alcohol/drug recovery while parenting an infant. The study explored women's experiences of recovery from alcohol/drug dependence during the first year after their infant's birth. Balancing emerged as the process that explained how women successfully integrated the recovery and motherhood processes into their identity. Understanding the balancing process will increase sensitivity of care. PMID- 10808624 TI - Parental encouragement, protection, and advocacy for Mexican-American children with chronic conditions. AB - Families face great challenges in caring for a child with a chronic condition, yet little is known about intrafamilial relationships, especially among Mexican Americans. An interpretive study, using symbolic interaction, with 25 adult informants, representing 19 mostly Mexican-American families, is reported. Findings include description of the process, "Keeping My Child Close to Me," by which parents, particularly mothers, provided protection, encouragement, and advocacy for their chronically ill child. Families were close and willing to jointly sacrifice to meet their special needs; however, some strains resulted from differences in perceptions between spouses and lack of attention to siblings during times of focus on the child with a chronic condition. PMID- 10808625 TI - Practical issues in obtaining child assent for research. AB - This article describes ways to engage children in clinical research through a carefully planned assent process. When assent is handled well, children may gain a sense of control over their choice to participate in research, which may in turn enhance their cooperation in the process. Ways to engage children in the assent process, the historical development of consent and assent, and discussion of the issues surrounding parental consent and child assent are presented. Practical suggestions are provided to make research participation a positive and appealing process for children. PMID- 10808626 TI - Systemic necrotizing vasculitis: a review, with the personal perspective of a nurse educator. AB - Although its definition is simple, systemic necrotizing vasculitis (SNV) is a complex, potentially life-threatening disease. By reviewing current published literature, this article provides an overview of vasculitis and focuses on what is common to the many individual systemic vascular diseases (vasculitides). A case history of a 10-year-old girl with SNV is included. Parse's Human Becoming theory is reviewed briefly as it applies to this case. PMID- 10808627 TI - Child mental health policy. PMID- 10808628 TI - The child in the community: nursing makes a difference. Counting children. PMID- 10808629 TI - Pediatric sedation: contributing to evidence-based practice. AB - Sedation of pediatric patients is a current practice issue; however, little has been published on performance measurement or evidence-based practice. The development of educational initiatives, the design of performance-improvement activities, and the validation of practice standards are described. Performance measures specific to the process of sedating pediatric patients are examined to show how to turn data into useful information to guide decision-making and nursing-practice changes. PMID- 10808630 TI - Multidisciplinary teams in the United Kingdom: problems and solutions. AB - Creating a team requires a clear strategy with objectives and boundaries. Teams succeed or fail depending on the enthusiasm and dedication of the members. Important points to consider in maintaining effectiveness include team philosophy, leadership, dynamics, communication, and workload. Members provide a unique contribution that must be valued and respected. Maintaining a productive and friendly team can become difficult. Issues such as uncertainty of the team's position in the overall service, caseload, poor coordination between team members, and uneven work distribution may cause disruption. To ensure a functioning and useful team, members must be constantly aware of their working relationships with each other and the impact they have on patients, families, and colleagues. PMID- 10808632 TI - Economic impact of hospital closure on small rural counties, 1984 to 1988: demonstration of a comparative analysis approach. AB - Hospital closure in a rural community may affect the locale's economic prospects as well as the health of its residents. Studies of economic effects have primarily relied on modeling techniques rather than observation of actual change. This study demonstrates the use of a comparative analysis approach for estimating the economic effects of hospital closure on small rural counties. The experiences of 103 small rural counties at which a hospital closed between 1984 and 1988 was compared with a matched group of counties at which no closure took place. "Comparable" counties were selected based on seven scales measuring the similarity between a closure county and potential comparisons. Three scales examined population and economic characteristics in the year before closure; two scales measured change throughout a three-year period preceding closure; and two scales measured change throughout a five-year period preceding closure. Closure effects were measured through a multivariate analysis of the post-closure economic history of closure and comparison counties. The key assumption is that similar counties should have similar experiences over time. If an event occurs within some of these counties but not others, this event should have visible effects. Comparative analysis suggested that earned income in closure counties (excluding farming and mining income) was lower than in comparison counties subsequent to closure and that labor force growth was similarly affected. A comparative analysis approach produces results that parallel those obtained from economic modeling and should be considered for further research. PMID- 10808631 TI - Cardiovascular disease risk factors and obesity of rural and urban elementary school children. AB - Previous studies on the influence of a rural/urban setting on the prevalence of cardiovascular disease risk factors in children have not sufficiently controlled for socioeconomic status, race, gender, and perhaps, may not have included a representative sample of rural and urban children. This study compared the cardiovascular disease risk factors and rate of obesity of children living in rural and urban settings. It also determined the magnitude of the effect of the rural/urban setting on cardiovascular disease risk factors and obesity when controlling for race, socioeconomic status, and gender. The subjects were 2,113 third- and fourth-grade children; 962 from an urban setting and 1,151 from a rural setting. Height, weight, skinfolds, resting blood pressure, and total cholesterol levels were measured. Aerobic power (pVO2max) was estimated from cycle ergometry. Physical activity and smoking history were obtained from a questionnaire. Clustering analyses using adjustment for sample error indicated that total cholesterol, blood pressure, smoking, and physical activity levels of rural and urban children were not different (P > 0.10); however, body mass index and sum of skinfolds was greater for rural youth (P < 0.004). Logistic regression indicated that rural children had a 54.7 percent increased risk of obesity (P = 0.0001). This study's results indicate that, in children, a rural setting is associated with obesity, but not with the major risk factors associated with cardiovascular disease. PMID- 10808633 TI - Dimensions of retention: a national study of the locational histories of physician assistants. AB - This study describes the locational histories of a representative national sample of physician assistants and considers the implications of observed locational behavior for recruitment and retention of physician assistants in rural practice. Through a survey, physician assistants listed all the places they had practiced since completing their physician assistant training, making it possible to classify the career histories of physician assistants as "all rural," "all urban," "urban to rural" or "rural to urban." The study examined the retention of physician assistants in rural practice at several levels: in the first practice, in rural practice overall and in states. Physician assistants who started their careers in rural locations were more likely to leave them during the first four years of practice than urban physician assistants, and female rural physician assistants were slightly more likely to leave than men. Those starting in rural practice had high attrition to urban areas (41 percent); however, a significant proportion of the physician assistants who started in urban practice settings left for rural settings (10 percent). This kept the total proportion of physician assistants in rural practice at a steady 20 percent. While 21 percent of the earliest graduates of physician assistant training programs have had exclusively rural careers, only 9 percent of physician assistants with four to seven years of experience have worked exclusively in rural settings. At the state level, generalist physician assistants were significantly more likely to leave states with practice environments unfavorable to physician assistant practice in terms of prescriptive authority, reimbursement and insurance. PMID- 10808634 TI - Consumer and provider evaluation of rural pharmacy services. AB - This paper compares consumer and provider perceptions regarding access to pharmacy services in rural Illinois, given a decrease in the number of pharmacies. Consumer data are from the Illinois Rural Life Panel in which more than 1,800 respondents answered questions about availability and use of pharmacy services and about insurance coverage and cost. A survey of all licensed retail pharmacies in 74 rural Illinois counties and in seven non-rural counties provided pharmacy background information and was the source of data on changes in profitability and payment sources. The data provided insight on factors that affect access. Descriptive statistics were used to analyze data from both groups to compare perceptions about access. The objective was to evaluate current access to pharmacy services and implications for future access from the perspective of consumers and pharmacists. Results from rural consumers show access is currently good; 77 percent have a local pharmacy, and 64 percent prefer this source. Future access is of more concern. Pharmacy survey results show 81.5 percent of rural pharmacies are experiencing declining profits from drug sales. Restricted reimbursements from third-party payers, demands of managed care and expanded competition are seen as threats to retention of local pharmacies and continued good access. An important finding, especially given survey evidence of increased managed care penetration, is the difference in views of pharmacists and consumers regarding the effects of managed care on access. Pharmacy survey data also revealed differences between rural and non-rural pharmacies. PMID- 10808635 TI - Can community organization strategies be used to implement smoking and dietary changes in a rural manufacturing work site? AB - A one-year intervention project was developed and implemented to demonstrate the utility of using community organization methods to mobilize a rural, predominantly minority work site community toward smoking and dietary change. This intervention for smoking and dietary change was conducted in a rural work site (n = 235 at baseline) and guided by employees. It involved activities to change the work site environment and the behaviors of individuals. A community advisory board (n = 15) made up of members of the work site was established, and it met monthly with members from the research team to design and implement nine cancer prevention activities that were targeted to the needs of this community. Activities and information were disseminated to the employees during a nine-month period. Surveys were administered prior to and following the delivery of the intervention. This project was successful in engaging a rural manufacturing work site community in thinking about cancer prevention strategies. Results of this intervention demonstrated significant increases in numbers of smoking cessation attempts, reported fruit and vegetable consumption, self-efficacy for dietary change and perceived risk for cancer. Work site social norms changed as evidenced by employee perceptions of co-worker support of dietary and smoking change (all ts > 1.95, all Ps < 0.05). Other results with marginal statistical significance (P < .015) but potentially useful for future studies include increased intentions to reduce the fat in the diet. In light of the low-intensity and time-limited nature of this community organization intervention, the observed changes in dietary and smoking behaviors are encouraging and support the use of these strategies in rural, culturally diverse work sites. PMID- 10808636 TI - What we say and what we do: county-level public spending for health care. AB - The purpose of this study is to examine county-level public spending for health care services in Kansas and to explain variation in spending levels with a model composed of population density, population age and per capita income. Data are abstracted from budget documents for all 105 counties in Kansas for the years 1994, 1995 and 1996. Health care expenditures are defined as county tax revenues spent for ambulance, hospitals, ambulatory care, home health services, nursing homes, and mental health and substance abuse services. Results show that Kansas counties spent between 12.1 percent and 13.6 percent of their budgets to fund local health care services between 1994 and 1996, spending more than $133 million in 1996 alone. In 10 counties, one-quarter to one-third of the budget went for health services. Low population density and relatively high per capita income explained nearly one-third of the variation in how much counties spent and an even greater proportion when analysis was limited to the most rural counties. Findings from this study suggest there may be a significant local commitment in the United States to publicly supported health care services, more support than typically recognized and perhaps more than is estimated in national health care spending data. Future research on the economic effects of the health sector on local communities should take account of local spending for health care, especially at the county level. PMID- 10808637 TI - Moderating effects of perceived control and need for clarity on the relationship between role stressors and employee affective reactions. AB - The authors examined the salience of perceived control and need for clarity as "buffers" of the adverse consequences of role stressors by using hierarchical regressions on role ambiguity and role conflict, with job satisfaction and psychological strain as the criterion variables. In a sample of U.S. and New Zealand employees, perceived control was directly associated with higher satisfaction and reduced strain but displayed no moderating effect on stressor outcome relationships. Need for clarity, on the other hand, was a significant moderator of the relationship of role ambiguity and conflict to both satisfaction and strain; that finding suggests that researchers could give more attention to dispositional variables in examining the correlates of role stressors. PMID- 10808638 TI - Controllability and stability in the self-serving attributions of sport spectators. AB - Spectators often attribute their athletic team's victories to internal causes and its losses to external causes (e.g., A. H. Hastorf & H. Cantril, 1954; R. R. Lau, 1984; L. Mann, 1974). This self-serving attributional pattern is most common among fans with a strong psychological attachment to their team (D. L. Wann & T. J. Dolan, 1994). The authors examined the relationships among identification, game outcome, and controllable and stable attributions. Their 1st hypothesis was that high-identification fans after a victory, compared with high-identification fans after a loss and low-identification fans after either outcome, would be more likely to exhibit self-serving attributional patterns by attributing their team's successes to controllable and stable causes. Their 2nd hypothesis was that high identification fans would be more likely than low-identification fans to attribute their team's successes to internal causes and its failures to external causes. U.S. college students high and low in identification first watched their university's men's basketball team win or lose a contest and then completed measures of identification and attribution. The results confirmed the hypotheses. PMID- 10808639 TI - Directing wayfinders with maps: the effects of gender, age, route complexity, and familiarity with the environment. AB - The participants were 360 Canadian undergraduates. After receiving written scenarios involving a campus visitor seeking direction to a nearby destination (simple route) or a distant destination (complex route), they drew maps to that destination. The authors varied the visitor's gender, age, and familiarity with campus. They analyzed the content of the students' maps in terms of cardial indicators (compass grid, correct north-south orientation), landmarks, labeled buildings, directional arrows, and supplemental written directions. The men tended to provide more cartographically complete maps than the women, though there were no gender differences in use of landmarks or labeled buildings. The men were significantly more inclined than the women to take visitor characteristics into account, providing more complete maps to visitors navigating complex routes, to newcomers, and to older visitors who were unfamiliar with the campus. The men had more confidence that their maps would successfully lead visitors to their destinations. Route complexity led to greater use of landmarks, labeled buildings, and cardinal indicators. The participants' self-reported familiarity with campus had little effect on their direction giving, except for a greater use of labeled buildings in maps. PMID- 10808640 TI - Cognitive organization of favorable and unfavorable beliefs about performing a behavior. AB - Intentions to perform behaviors are influenced by beliefs about their consequences, but researchers have not addressed how such beliefs are cognitively organized. In 3 experiments, the authors tested the hypothesis that beliefs are organized according to whether they are favorable or unfavorable in regard to performing a behavior. In Experiment 1, a group of U.S. students first read a list of beliefs favorable and unfavorable to going to a vacation resort and then listed their beliefs about condom use. In Experiment 2, another group of U.S. students listed their beliefs about condom use. In Experiment 3, a 3rd group of U.S. students listed their beliefs about a novel behavior (asking the experimenter for candy). The results of all 3 experiments were consistent with the hypothesis that people cognitively organize their beliefs according to whether they are favorable or unfavorable to the behavior in question. PMID- 10808641 TI - Psychological involvement in parenthood among Ukrainians. AB - In a sample of 100 parents in Ukraine, the authors examined variables that affect the extent of psychological engagement in the role of parenthood. The participants completed questionnaires about their beliefs and expectations as parents. As predicted by the triangle model of responsibility (B. R. Schlenker, 1997; B. R. Schlenker, T. W. Britt, J. W. Pennington, R. Murphy, & K. J. Doherty, 1994), 3 variables significantly predicted psychological engagement: task clarity (i.e., a clear understanding of the duties of a parent), personal obligation (i.e., the duty to follow prescriptions for being a good parent), and personal control (i.e., personal control over aspects of child rearing). In addition, task clarity and personal obligation predicted the participants' expectations of the quality of their children's futures; however, personal control did not, perhaps because of contemporaneous economic and political instability in Ukraine. PMID- 10808642 TI - The effect on expected job success of the connotative meanings of names and nicknames. AB - The authors conducted this study among U.S. students to determine whether expectations of job success are related to the degree of match between the masculinity or femininity of men's or women's names and the stereotypic masculinity or femininity of planned occupations. Results confirmed the predictions: The greater the match among (a) the gender of names, (b) the participants' ratings of the masculinity or femininity of those names, and (c) the masculinity or femininity of planned occupations, the greater the expected likelihood of job success. PMID- 10808643 TI - Self-concepts of hospitalized and day-care psychiatric patients. AB - The authors administered questionnaires to 44 hospitalized and 55 day-care psychiatric patients in Hong Kong. The groups were similar in sex, age, and education. The hospitalized participants, compared with the day-care participants, showed significantly higher self-concepts in general as well as in the particular aspects of social and personal aspirations. The findings indicate that support at the community level is important to help the psychiatric patients' transition from hospital care to day care. PMID- 10808644 TI - Culture and self in South Africa: individualism-collectivism predictions. AB - People from collectivist cultures may have more concrete and interdependent self concepts than do people from individualist cultures (G. Hofstede, 1980). African cultures are considered collectivist (H. C. Triandis, 1989), but research on self concept and culture has neglected this continent. The authors attempted a partial replication in an African context of cross-cultural findings on the abstract concrete and independent-interdependent dimensions of self-construal (referred to as the abstract-specific and the autonomous-social dimensions, respectively, by E. Rhee, J. S. Uleman, H. K. Lee, & R. J. Roman, 1995). University students in South Africa took the 20 Statements Test (M. Kuhn & T. S. McPartland, 1954; Rhee et al.); home languages were rough indicators of cultural identity. The authors used 3 coding schemes to analyze the content of 78 protocols from African language speakers and 77 protocols from English speakers. In accord with predictions from individualism-collectivism theory, the African-language speakers produced more interdependent and concrete self-descriptions than did the English speakers. Additional findings concerned the orthogonality of the 2 dimensions and the nature and assessment of the social self-concept. PMID- 10808645 TI - Academic performance and perceived validity of grades: an additional case for self-enhancement. AB - The authors investigated the role of academic self-esteem and academic performance in U.S. college students' perceptions of the validity of their grades (overall grade point average [GPA]). A sample of 208 (80 male, 128 female) college students completed a survey that included an academic self-esteem scale and a measure of the perceived validity of grades. The authors assessed academic performance level by the participants' actual overall GPAs. The results of a hierarchical multiple regression analysis supported the weak form of self enhancement theory (J. S. Shrauger, 1975). Thus, regardless of their self-esteem levels, the students with higher GPAs, compared with those with lower GPAs, tended to see the overall GPA as a more valid indicator of academic ability. PMID- 10808646 TI - Children's conceptions of characteristic features of category members. AB - The authors investigated children's conceptions of the characteristic features of category members and found that their conceptions underwent qualitative developmental changes. They hypothesized that (a) children initially tend to perceive category members in terms of individual characteristics (i.e., internal dispositions and behavioral patterns) and (b) only later do they conceive of category members in terms of their shared beliefs and values. They explored these hypotheses in 2 studies: In Study 1, they investigated how Protestant and Catholic children in Northern Ireland understood the religious intergroup context in their own country; in Study 2, children in Italy were presented with a fictional scenario of intergroup conflict and asked to explain the causes of the conflict. The results of both studies confirmed the 2 hypotheses. PMID- 10808647 TI - Gender and context differences in alcohol expectancies. AB - In 2 studies, the authors investigated gender and context effects of alcohol expectancies among U.S. college students. In Study 1, they examined the responses of 771 women and 493 men to the Sexual Enhancement (SE) subscale and the Relaxation and Tension Reduction (TR) subscale of the Alcohol Expectancy Questionnaire (S. A. Brown, B. A. Christiansen, & M. S. Goldman, 1987). They found no gender differences when they controlled for drinking quantity. When they controlled for drinking frequency, the men scored significantly higher on the TR subscale than the women did. In Study 2, also among U.S. college students, 41 women and 21 men first imagined that they were on blind dates or in long-term relationships and then completed the SE and TR subscales. On the SE subscale, the women scored higher in the relationship context than in the blind-date context, whereas the men did not vary between contexts. All the participants also reported more tension-reduction expectancies in the relationship context than in the blind date context. PMID- 10808648 TI - Mail surveys of faculty and acquaintances-of-the-researcher bias. PMID- 10808649 TI - Gender role typing and attachment to parents and peers. PMID- 10808650 TI - The effect of student involvement on the development of academic self-concept. PMID- 10808651 TI - "Foot-in-the-mouth" versus "door-in-the-face" requests. PMID- 10808652 TI - Effect of group identification on the use of attributions. PMID- 10808653 TI - Caution: women at work. PMID- 10808654 TI - Perspectives on women's occupational health. AB - This commentary provides a brief history of the development of the field of women's occupational health and safety. It discusses the broadening of the field from one with a reproductive health focus to one that considers women in their many roles and varied stages of life. Much of this widening of focus was contemporaneous with the growth of the women's movement. The paper emphasizes that the work burden borne by employed women in general includes responsibilities for homemaking and caregiving in addition to paid employment. Possible implications of these multiple roles are discussed, as are the major social, physical, biological, and chemical hazards encountered in traditional female employment. A research agenda for overcoming some of the research and policy barriers to a complete understand of women's occupational health burden and to development of preventive policies is proposed. PMID- 10808655 TI - Ergonomic studies provide information about occupational exposure differences between women and men. AB - This article reviews published information and ergonomic analyses of work activity in several jobs usually done by women in order to help researchers and clinicians determine the extent to which women's health problems can be attributed to the workplace and where to intervene. Women in the workplace are often exposed to different musculoskeletal and psychological constraints, even when they share job titles with men. Job and task assignments may differ by sex, and gender-associated biological and social characteristics may interact with workplace constraints to modify the exposures or the effects of working conditions. Jobs preferentially assigned to women may have specific exposures that escape the attention of researchers. PMID- 10808656 TI - Working women and stress. AB - Occupational stress is a growing problem in US workplaces and may be a problem of particular magnitude for working women, in part because of sex-specific job stressors (sex discrimination and difficulties combining work and family). Although such stressors have received little research attention until recent years, new research indicates that these stressors may have a negative impact on health and well-being above and beyond the effects of general job stressors (work overload, skill underutilization, etc). A number of stress-reduction strategies have been shown to be useful for working women, ranging from the more common individual stress management techniques to higher-level interventions focused on removing the sources of occupational stress. This article provides a brief overview of occupational stress as it affects working women and presents research on approaches for reducing the negative effects of job stress. PMID- 10808657 TI - Managing patients with concerns about workplace reproductive hazards. AB - OBJECTIVES: To find out who uses an occupational reproductive consultation service, what proportion of patients have different types of workplace exposures, and what hypotheses can be generated about barriers to implementing medically necessary job modifications to promote reproductive health. METHODS: A case series study was conducted by reviewing medical records at two occupational health clinics. RESULTS: 51 patients (1 man and 50 women) were seen, 10 of whom wished to discuss a future pregnancy and 41 of whom were pregnant. Pregnant women worked with a mean of 15.5 different chemicals, and patients were also concerned about ionizing radiation, biological hazards, electromagnetic fields, and ultraviolet light. Pregnant women made clinic visits at a mean gestational age of 10.9 weeks. CONCLUSIONS: Only one man used the service, suggesting a lack of knowledge about possible paternal contributions to adverse reproductive outcomes. Many pregnant women visited the clinic too late to prevent harm from exposure to some teratogens, so preconception counseling may be of benefit. Cases are presented that illustrate ways in which the primary care provider can assist the patient who may be exposed to reproductive hazards. PMID- 10808658 TI - Video display terminals and miscarriage. AB - Concern about possible reproductive effects of using video display terminals (VDTs) was stimulated by reports of clusters of adverse pregnancy outcomes among groups of women who used VDTs. Exposure to electromagnetic fields emitted from VDTs was suspected of increasing the risk of miscarriage. Ten epidemiological studies have examined associations between VDT use and miscarriage, most of which were not originally designed to examine this hypothesis and suffer from methodological limitations. We review these studies and present previously unpublished data in support of our conclusion that for most women in modern offices, work with VDTs does not increase their exposure to electromagnetic fields or increase their risk of miscarriage. The miscarriage risk for women who work at high-stress jobs or with older, high-emission VDTs (ELF > 3 mG), however, is still uncertain. PMID- 10808659 TI - Women in construction: occupational health and working conditions. AB - Construction is one of the largest industries in the United States, employing 7.6 million workers, or about 5% of the US work force. More women have taken jobs in the construction industry over the last two decades, as they have in other nontraditional industries. In 1997, there were 8.1 million construction workers, of whom 781,000 (9%) were women. Approximately 2% of those were employed as skilled tradeswomen. There is no disputing that construction work is dangerous. Seventeen percent of all fatal on-the-job injuries occur in construction, which is about three times its 6% share of total employment. In this paper, we review the medical literature on the safety and health hazards for women working in the construction industry. Women have a different pattern of fatal injuries and some differences in patterns of nonfatal injuries than men and report unique problems and concerns related to working in this industry. PMID- 10808660 TI - Sex differences in principal farm operators' tractor driving safety beliefs and behaviors. AB - OBJECTIVE: To examine the widely accepted hypothesis that farm women are more concerned with safety issues and behaviors than their male counterparts are. METHODS: A telephone survey was administered to a random sample of Kentucky principal farm operators, 90 of whom were women. Participants were questioned about their tractor safety beliefs and practices. RESULTS: No significant sex differences in tractor safety perceptions and behavior were observed. CONCLUSIONS: Socialization of women to the role of principal farm operator may override their typically greater sensitivity to safety issues, an important consideration when designing safety campaigns for this population. PMID- 10808661 TI - From maternity to parental leave policies: women's health, employment, and child and family well-being. AB - Pregnancy and maternity are increasingly viewed as social as well as individual risks that require health protection, employment protection and security, and protection against temporary loss of income. Begun more than a century ago in Germany, paid and job-protected maternity leaves from work were established in most countries initially out of concern for maternal and child physical health. Beginning in the 1960s, these policies have expanded to cover paternity and parental leaves following childbirth and adoption as well. Moreover, they have increasingly emerged as central to the emotional and psychological well-being of children as well as to the employment and economic security of their mothers and fathers. They are modest social policies, but are clearly an essential part of any country's child and family policy. No industrialized country today can be without such provision, and the United States is a distinct laggard in these developments. PMID- 10808662 TI - Sharing the mantle of primary female care: physicians, nurse practitioners, and physician assistants. AB - OBJECTIVE: To examine the role that nurse practitioners (NPs) and physician assistants (PAs) play in women's health care as part of a larger study assessing the use of NPs and PAs as primary care practitioners. METHODS: We conducted qualitative key informant interviews with providers and administrators at nine managed care organizations and multispecialty clinics. RESULTS: Respondents indicated that although there were a number of reasons these institutions began to hire NPs and PAs, the shortage of women health care providers was an important contributing factor. Many women patients prefer to see same-sex providers, but there are not enough female physicians to meet this demand. NPs and PAs were more interested in preventive care than physicians were. For these reasons, NPs and PAs came to play a central role in the delivery of women's primary care within these institutions. CONCLUSION: Although the number of female physicians is increasing, there is no indication that the importance of NPs and PAs is waning. Rather, they have become valued members of the health care team. Women physicians will most likely be expected to provide primary care in teams with NPs and PAs. PMID- 10808663 TI - JAMWA and Latina Magazine collaborate on complementary and alternative medicine survey. PMID- 10808665 TI - Are there any more Louisa Burnses out there? PMID- 10808664 TI - The role of government in postgraduate training programs in women's health. AB - This paper examines the role of the federal government in postgraduate training programs in women's health. Health care professional education has long been an interest of many agencies within the Department of Health and Human Services. Although federal agencies have not been directly involved in the creation of curriculum or funding of training programs in women's health, they have helped to catalyze wide public and private support of these programs. The Office on Women's Health within the Department of Health and Human Services, the National Institutes of Health Office of Research on Women's Health, and the Health Resources and Services Administration have each provided recommendations for the inclusion of women's health in the medical school curriculum. Additionally, the Council on Graduate Medical Education, an advisory body to the Secretary of Health and Human Services and Congress, has recommended that physicians develop a broad understanding of health conditions specific to women. Although federal funding is not available for the development of training programs in women's health, continued public and private interest will foster the development of post graduate training programs that will improve the delivery of health care for women in the United States. PMID- 10808666 TI - National study of the impact of managed care on osteopathic physicians. AB - The study reported here was designed to provide insight into the impact managed care has had on osteopathic physicians' ability to practice medicine, as well as data to substantiate the prevalence of the specific problems encountered by the 40,000 osteopathic physicians in the United States. New data on the extent to which osteopathic physicians use osteopathic manipulative treatment was also obtained, as a review of the literature revealed only two previous surveys on the use of osteopathic manipulative treatment. The American Osteopathic Association hired an independent research company to conduct the survey. PMID- 10808667 TI - Use of magnetic resonance imaging to diagnose common wrist disorders. AB - Magnetic resonance imaging is being used more frequently to diagnose and plan treatment of wrist disorders. This article reviews the common pathologic lesions of the wrist: avascular necrosis, triangular fibrocartilage complex tears, ligamentous tears, ganglion cysts, carpal tunnel syndrome, and osteoarthritis. The typical magnetic resonance imaging characteristics of these lesions is discussed. PMID- 10808668 TI - Hurthle cell adenoma diagnosed by core needle biopsy in a male patient. AB - Hurthle cell adenomas (HCAs) are a rare and potentially lethal variant of follicular tumors of the thyroid. Considerable controversy exists regarding potential risk factors, diagnosis, and treatment of HCAs. The authors report the case of a 38-year-old male patient with an 8.3 cm x 3.5 cm HCA. Diagnosis was made preoperatively from a core needle biopsy and confirmed postoperatively on frozen section. Treatment consisted of a right lobectomy. PMID- 10808669 TI - Relationship between academic achievement and COMLEX-USA Level 1 performance: a multisite study. AB - The purpose of this study was to examine the relationship of performance on the Comprehensive Osteopathic Medical Licensing Examination (COMLEX-USA) Level 1 with academic performance at colleges accredited by the American Osteopathic Association (AOA). Eighteen (95%) of 19 AOA-accredited colleges and 2146 students (91% of those taking the June 1999 examination) met criteria and participated. Students were classified by school representatives on the basis of academic performance in the first 2 years of the curriculum. The relationships of Level 1 performance with assigned classifications and grade point averages (GPAs) were studied. Of students classified in the highest 20% academically, the Level 1 pass rate was 100%, with a mean score of 599. Of students classified in the lowest 5%, the pass rate was 63.5%, with a mean of 416.3. For 16 schools that provided GPAs, the within-school correlations between Level 1 scores and GPAs ranged from r = 0.76 to r = 0.85, with a mean correlation of r = 0.79. School representatives were also asked to indicate, for each student, whether they expected the student to pass the examination. Pass rate for students in the "sure pass" category was 98.9%; "borderline," 82.5%; and "concerns," 61.5%. Academic performance in the first 2 years of osteopathic medical school was strongly associated with performance on COMPLEX-USA Level 1. The national pass rate for this examination was similar to those in previous years, and it remains unclear why school representatives overpredicted the number of failures. Further research is needed. PMID- 10808670 TI - Early research: the A.T. Still Research Institute and Louisa Burns. PMID- 10808671 TI - His eighty-five years. 1913. PMID- 10808672 TI - A home for research. 1913. PMID- 10808673 TI - Arkansas Health Care Access Foundation Inc. PMID- 10808674 TI - Initial disease modifying antirheumatic drugs and prednisolone prescriptions for patients with rheumatoid arthritis: a 15-year study. AB - OBJECTIVE: To compare patterns and time trends of initial disease-modifying antirheumatic drugs (DMARDs) and prednisolone prescriptions for patients with rheumatoid arthritis (RA) by the rheumatologists at King Chulalongkorn Memorial Hospital, Bangkok, Thailand over a 15-year period, as well as their side effects. METHOD: Medical records of all patients with RA seen at the Rheumatology Clinic from January 1983 to June 1997 with a duration of follow-up of 6 months or more were reviewed. Information on the disease, initial DMARDs prescriptions and their side effects, prednisolone use, dosage and side effect(s) were focused and compared among three 5-year periods (1983-1987, 1988-1992 and 1993-1997). RESULTS: 236 patients were included in this study. There were 44, 82 and 110 patients in the first, second and third period, respectively. Methotrexate (MTX) was the most frequently prescribed DMARD in all time periods. Dapsone and intramuscular (i.m.) gold were prescribed in the first period while antimalarial drugs and sulfasalazine (SSZ) were increasingly used in the second and third periods. Combination treatment of DMARDs was first used in the third period. Side effects from MTX were observed in patients with a longer duration of treatment (p < 0.05). Patients prescribed combined DMARDs did not develop more side effects compared with those who had monotherapy. Prednisolone was prescribed in 57.2 per cent of the patients, most being newly prescribed at the clinic. Mean starting dose of prednisolone was 8.9 mg per day. 64 patients took prednisolone together with non-steroidal antiinflammatory drugs (NSAIDs). Gastrointestinal side effects did not increase in these patients. CONCLUSION: MTX was the most frequently prescribed DMARDs regardless of the time period. Antimalarial drugs, SSZ and combination of DMARDs (most were MTX + chloroquine) have been prescribed more in the last 5 years, while dapsone, auranofin and i.m. gold were rarely used as initial DMARDs. Low dose prednisolone was prescribed in more than half of the patients with RA. Side effects from DMARDs and prednisolone found in this study were comparable to previous reports. PMID- 10808675 TI - Captopril suppression tests in normotensive individuals. AB - OBJECTIVE: We aimed to measure the serial (time related) concentrations of aldosterone and renin after a single dose of 25 mg of captopril in normotensive individuals. With these results, we aim to ascertain the best sampling times for future tests. DESIGN: Six normotensive subjects were studied under two different conditions--once in a supine position and once under normal (erect-position) activity. Each subject was given 25 mg of captopril and his serum aldosterone and renin concentrations measured at half hourly intervals for four hours. Simultaneous half hourly blood pressure measurements were also noted. RESULTS: All subjects showed suppression of aldosterone levels with ingestion of captopril, and in the supine position maximal suppression was consistently observed from the 3rd to the 4th hour after ingestion. When studied in the erect posture, although there was significant suppression, this was not sustained, and a paradoxical rise of aldosterone levels was seen after 2 hours. Renin activity measured during this study showed no consistent patterns. The renin levels were unchanged in almost 40 per cent of cases, and raised in about 60 per cent of cases. None of the subjects had symptomatic postural hypotension after 25 mg of captopril. CONCLUSIONS: The captopril suppression test is safe and effective in assessing the suppressibility of aldosterone and the period of maximal suppression is from the 3rd to the 4th hour after oral ingestion of captopril. PMID- 10808676 TI - Eradication of Helicobacter pylori with lansoprazole based triple therapy in peptic ulcer disease. AB - Lansoprazole 30 mg, amoxicillin 1000 mg, and tinidazole 500 mg were given twice daily to 39 peptic ulcer patients (26 duodenal and 13 gastric ulcer, mean age 52.4 +/- 15.01) who had H. pylori infection for two weeks. Additional lansoprazole 30 mg daily was given to duodenal and gastric ulcer patients for another two and six weeks respectively. Follow-up gastroduodenoscope was performed at fourth and eighth week and eighth and twelfth week for all duodenal and gastric ulcer patients, respectively. H. pylori status was evaluated by rapid urease test (CLO test) and histology at first and last endoscope. The ulcers were healed at the last endoscopy in 11 (85%) gastric ulcer patients and 24 (92%) duodenal ulcers patients. H. pylori infection was eradicated in 31 patients (79%). Mild side effects were observed in 15 per cent. In conclusion, 2 week regimen of lansoprazole, amoxicillin, and tinidazole triple therapy resulted in a relatively high healing rate of peptic ulcer (90%) and an acceptable eradication rate of H. pylori infection (79%). PMID- 10808677 TI - Five-year follow-up of prostate cancer in Siriraj Hospital. AB - OBJECTIVE: To determine the clinical features, tumor characteristics, and treatment outcomes of Thai men with prostate cancer. MATERIAL AND METHOD: We retrospectively evaluated the clinical features, tumor characteristics, and treatment outcomes of 95 patients who were registered in Siriraj Hospital from 1993 to 1995. A survival end point in each stage was determined. RESULTS: The mean age was 72.37. The distributions of stage were 7.5 per cent for stage A, 1.1 per cent for stage B, 67.7 per cent for stage C, and 23.7 per cent for stage D. The prognosis of a clinical localized disease appeared good. Most patients with a urinary symptom were highly associated with stage C or stage D disease and were treated by hormonal therapy. With a maximal follow-up of 60 months, the median survival of stage C and D patients was 45 and 12 months, respectively. CONCLUSION: Most Thai patients with prostate cancer were older than the life expectancy of Thai men. They presented with urinary symptoms and had locally advanced or advanced disease. With hormonal treatment, their prognoses were not impressive. PMID- 10808678 TI - Bone mineral density in primary and secondary amenorrhea. AB - Amenorrhea in young women is one of the best clinical indicators for estrogen deficiency, except in the presence of gynecological structural pathology. This study aimed at investigating bone mineral density (BMD) in patients with primary and secondary amenorrhea. Thirty-six patients were enrolled in the study, seven with primary amenorrhea (mean age 24.3 +/- 4.5 yrs.) and twenty-nine with secondary amenorrhea (mean age 31.1 +/- 6.9 yrs.). Eighteen regularly menstruating women (mean age 31.8 +/- 3.7 yrs.) served as controls. BMD was measured at lumbar spine, femoral neck, Ward's triangle and trochanter. RESULTS: BMD was significantly decreased in both primary and secondary hypoestrogen amenorrheic patients. Primary amenorrheic patients were more severely affected with a BMD mean Z score below 80 per cent (osteopenia) at all sites measured. The age of primary amenorrheic women also strongly correlated with degree of demineralization. This should emphasize the importance of early diagnosis and treatment of young amenorrheic patients. PMID- 10808679 TI - Photodynamic therapy in management of head and neck cancers and precancerous lesions. AB - Photodynamic therapy (PDT) is a new form of cancer treatment with low morbidity. In this study, PDT was evaluated for its effectiveness in management of recurrent or widespread precancerous lesions, primary cancers in inoperable sites, recurrent or residual cancers which were refractory to radiotherapy and chemotherapy, and advanced tumors in the head and neck. Fifty-one patients were treated over a period of 5 years. A 91.67 per cent complete response rate was observed for T1 tumors (primary and recurrence) with a recurrent rate of 27.27 per cent. Nasopharyngeal carcinoma was highly responsive to PDT since all T1 and T2 tumors responded completely. This was in contrast to cancers in the soft palate which failed in most cases possibly due to inadequate light dose distribution. PDT was remarkably effective in curing premalignant diseases (100% complete response rate). Postoperative PDT was equally effective in treating the microscopic residual malignancy. For advanced tumors, PDT in adjunct to conventional modalities could induce complete response in 5 out of the 10 patients and resolve symptoms in 4 cases. The mean follow-up time for this series was 28.3 months (range 3-66 months). In conclusion, PDT is a useful modality for the treatment of head and neck tumors and precancerous lesions presenting in forms or under conditions that posed considerable difficulties in management by conventional approaches. PMID- 10808680 TI - Correlation between some discrimination functions and hemoglobin E. AB - The most widely used discriminant functions and red cell indices for differential diagnosis of thalassemia traits from iron deficiency anemia were evaluated for their abilities to identify HbE-containing blood samples. The functions were as follows: F1 = 0.01 x MCH x (MCV)2; F2 = RDW x MCH x (MCV)2/Hb x 100; F3 = MCV/RBC; and F4 = MCH/RBC. Other red cell parameters including RDW, hemoglobin content, mean cell volume, mean cell hemoglobin as well as red cell counts, were also evaluated to distinguish HbE from the normal population. Hemoglobin electrophoresis was used as a confirmatory test. The results showed that F1, F2 and F3 as well as other red cell parameters of HbE-containing samples were different from those of HbA2A-containing red cells although there was no statistical significance. However, F4 and MCHC showed no difference between the two groups. It can be concluded from the present study that identification of hemoglobin E especially the heterozygous form by using parameters from an electronic cell counter is not easy. Discriminant functions and red cell indicies might be used as an initial diagnosis. But confirmation is needed in all cases. Applying the MCV of 80 fl will miss 5 per cent of hemoglobin E carrier but will not miss the homozygous form. PMID- 10808681 TI - Study of plasma hormones and lipids in healthy elderly Thais compared to patients with chronic diseases: diabetes mellitus, essential hypertension and coronary heart disease. AB - Simultaneous measurements of serum estradiol, testosterone, cortisol, prolactin, total cholesterol (TC), high density lipoprotein cholesterol (HDLC), low density lipoprotein cholesterol (LDLC) and triglycerides in Thai men and postmenopausal women aged over 50 years were carried out in four groups of subjects: healthy controls, and patients with essential hypertension, non-insulin dependent diabetes mellitus (NIDDM), and coronary heart disease. Hypertriglyceridemia and hypercholesterolemia were found more often in patients with essential hypertension than in patients with other diseases. Low levels of HDLC with high TC/HDLC and LDLC/HDLC ratios occurred more frequently in coronary heart disease patients. Hypertensive men had the highest plasma estradiol levels while men with coronary heart disease had the least testosterone levels compared with men with the other two diseases. Decreased testosterone and/or increased estradiol may have an adverse effect on lipid profiles in elderly men. However, neither the sex hormones, cortisol, nor prolactin, appeared to have any influence on serum lipids and lipoproteins in elderly women. These findings in the Thai population are consistent with those previously reported in other populations. PMID- 10808682 TI - Seizure threshold in ECT: II. Dose titration vs age and half age methods. AB - Seizure threshold determination is of crucial importance in optimizing electrical stimulus dosage at electroconvulsive therapy (ECT). We measured initial seizure threshold by means of Srinakharinwirot University titration schedule in 106 patients with schizophrenia or schizoaffective disorder, receiving bilateral ECT. Seizure threshold was approximately 106 millicoulombs on average, and varied 5 fold across patients. Seizure threshold was directly related to age, but inversely related to motor seizure duration. Comparisons of stimulus charge were done with the Age and Half age methods. By using the Half age method, 68 per cent of patients would have seized at the first stimulation and resulted in a closer mean charge to dose-titration method than the Age method. The results may have important clinical implications for stimulus dosing strategy in ECT. PMID- 10808683 TI - Charts of Thai fetal biometries: 1. Methodology. AB - A cross-sectional study was conducted in order to construct new reference charts for Thai fetal biometries that are commonly used in obstetric ultrasound practice. We discussed and illustrated a sound appropriate study design and statistical analysis which lead to more valid results. A total of 621 normal pregnant women between 12-41 weeks of gestation and their fetuses were recruited. Each fetus was measured once at a randomly assigned gestational age specifically for the purpose of this study only. Stepwise linear regression technique was used to model the mean and its standard deviation as functions of gestational age. Goodness of fit and normality of the data were checked before the final models were chosen. Reference centiles were derived, taking into account the increasing variation as pregnancy proceeds. We demonstrated the stated technique with humerus data from the same study. Reference charts for other fetal biometries have been derived and are presented in subsequent papers. PMID- 10808684 TI - Charts of Thai fetal biometries: 2. Biparietal diameter. AB - A cross-sectional study was conducted in order to construct a new reference chart for Thai fetal biparietal diameter (BPD). A total of 621 normal pregnant women between 12-41 weeks of gestation and their fetuses were recruited. Measurements were made once at a randomly assigned gestational age specifically for the purpose of this study only. Due to unfavorable fetal position in some cases, BPD data were available in 613 measurements. Linear regression models were fitted separately to estimate the mean and standard deviation as functions of gestational age. Reference centiles were constructed from both equations, assuming the data were normally distributed. A new reference centiles chart for BPD is presented and compared with previously published data. Our derived centiles were clearly lower than those from Western studies showing the importance of racial differences between populations. This elucidates the need to develop fetal biometries charts specifically for each region. PMID- 10808685 TI - Charts of Thai fetal biometries: 3. Femur length. AB - A cross-sectional study was conducted in order to construct a new reference chart for Thai fetal femur length (FL). A total of 621 normal pregnant women between 12 41 weeks of gestation and their fetuses were enrolled. Measurements were made once at a randomly assigned gestational age specifically for the purpose of this study only. Femur length data were available in 608 measurements due to unfavorable fetal position in some cases. Linear regression technique was used to model separately the mean and standard deviation as functions of gestational age. Reference centiles were constructed from a combination of both models, assuming the data were normally distributed. A new reference centiles chart for FL is presented and compared with previously published data. While our derived centiles were clearly lower than those from Western studies, they were found comparable with those from a Thai study. This demonstrated the important effect of racial differences between populations on fetal biometries and elucidates the need to develop fetal biometries charts specifically for each region. PMID- 10808686 TI - Charts of Thai fetal biometries: 4. Abdominal circumference. AB - A cross-sectional study was conducted in order to construct a new reference chart for Thai fetal abdominal circumference (AC). A total of 621 normal pregnant women between 12-41 weeks of gestation and their fetuses were recruited. Measurements were made once at a randomly assigned gestational age specifically for the purpose of this study only. Due to unfavorable fetal position in some cases, AC data were available in 615 measurements. Linear regression models were fitted separately to estimate the mean and standard deviation as functions of gestational age. Reference centiles were constructed from both equations, assuming the data were normally distributed. A new reference centiles for AC is presented and compared with previously published data. Our derived centiles were lower than those from Western studies which may partly be due to racial differences. This emphasizes the need to develop fetal biometries charts specifically for each region. PMID- 10808687 TI - Long-term follow-up study of IgM associated nephrotic syndrome patients: clinical outcome and prognostic indicators. AB - RATIONALE: Even though it is the most common primary glomerular disease, the natural history and prognosis of IgM associated nephrotic syndrome have not been well established. OBJECTIVES: To determine the (1) responsiveness to prednisolone therapy, (2) long-term clinical and laboratory outcomes, and (3) prognostic indicators to prednisolone therapy in patients with IgM associated nephrotic syndrome. STUDY DESIGN: Clinical descriptive, longitudinal study. SUBJECTS: Seventy two biopsy-proved IgM associated nephrotic syndrome patients, diagnosed between 1978-1996 at Vajira Hospital, Bangkok, were included in the study. METHOD: Clinical parameters with age, sex, duration of edema, blood pressure and laboratory findings such as hematuria, BUN, creatinine, albumin, and cholesterol, 24-hour urine protein, and stool examination, were collected pre-renal biopsy. Each patient was treated with 45-60 mg of prednisolone according to body weight, for up to 8 weeks. Each patient was followed-up every 4-weeks for clinical and laboratory evaluations, and for adjusting the steroid dosage. Clinical responses were stratified into 3 groups as steroid responsive (SRP), steroid dependent (SD), and steroid resistant (SRS). MAIN OUTCOME MEASURES: (1) Frequency and types of steroid responsiveness. (2) Incidence of hypertension, hematuria, renal insufficiency, end-stage renal disease, and survival during the follow-up. (3) Prognostic indicators for initial clinical response to prednisolone, and for long term morbidity and mortality. RESULTS: Forty eight of the 72 patients (66.67%) were responsive to prednisolone at 8-weeks, the 24 remaining patients (33.33%) were nonresponsive. High proteinuria of 7.66 +/- 4.14 g/D was the only good prognostic indicator to initial prednisolone therapy (p < 0.03). During the follow-up, 42(58.33%), 26(36.11%), and 4(5.56%) patients were SRP, SD, and SRS, respectively. There were no prognostic indicators associated with long-term steroid responsiveness. Of the 60 patients followed-up for more than one year; 34, 15 and 11 patients were followed-up for 1-5, > 5-10 and > 10 years, respectively. Hematuria and proteinuria were more frequent among the SRS group (p < 0.01 and 0.02, respectively) during the follow-up. Only one patient, initially in the SD group, and later on became SRS, died. CONCLUSION: Patients with IgM associated nephrotic syndrome had very good response to prednisolone therapy. It had a very slow progressive course, with low morbidity and mortality. PMID- 10808688 TI - Dengue infection with unusual manifestations: a case report. AB - Since 1978, there has been an increasing number of reported cases of dengue infection with unusual manifestations and most of them had dengue shock syndrome. We report here one patient who had dengue hemorrhagic fever grade II with liver failure and hepatic encephalopathy and very high elevation of liver enzymes. She made a complete recovery after conservative therapy. She is the fourth case of reported dengue hemorrhagic fever grade II who had unusual manifestation. PMID- 10808689 TI - The Helsinki Declaration regarding ethics in clinical research. PMID- 10808690 TI - Compliance with the law on car seat-belt use in four cities of Thailand. AB - Surveys to determine the scope of compliance with the law requiring seat-belt use in Thailand were conducted by observation and interviews with drivers in four cities: namely, Bangkok Metropolis, Chiang Mai, Phuket and Nakhon Ratchasima. The work was carried out in two separate sessions: during the first month following enactment of the law, and six months after its enactment. The sample comprised 46,949 vehicles in the first session (January 1996) and 76,188 vehicles in the second session (July 1996). The results showed that 42.7 per cent of drivers used seat-belts in January and 30.7 per cent in July. When the data were disaggregated according to cities, it was found that more Bangkok drivers complied with the seat-belt law than in Phuket, 24.6 per cent; Chiang Mai, 22.1 per cent; and Nakhon Ratchasima, 18.3 per cent relatively low compliance rate was encountered among drivers of pick-up trucks (including those with modified roofs) and vans, particularly farmers and the self-employed. Women drivers tended to abide by the law more often than men (OR = 1.7, 95% CI = 1.12, 1.23). Inter-city travelers wore seat-belts more than those traveling in the city (OR = 1.74, 95% CI = 1.68, 1.80). About one-fifth of non-users or those who rarely used seat-belts were unsure of the effectiveness of seat-belts in preventing serious injury or death in the case of an accident. Discomfort associated with using seat-belts and the perception that they were rendered unnecessary because of slow traffic in cities were other reasons for non-compliance in 50.6 per cent and 43.9 per cent of cases, respectively. The decline in compliance with the law six months after its enactment indicates that there may be a problem with uniform and consistent enforcement of the law. PMID- 10808691 TI - Validity of clinical use of the clock-drawing test in Thai elderly patients with memory problems. AB - Forty eight elderly Thai patients, 12 demented, 36 non-demented who consecutively attended a geriatric clinic, at Chulalongkorn University Hospital, with a memory problem were recruited in the study which aimed to determine the validity of the clock-drawing test (CDT) and Chula mental test (CMT). Subjects took the Chula Mental Test (CMT) and were asked to draw a clock on a preprinted 12 centimeters circle showing the time of 11:10. Clocks were scored using the Chula Clock drawing Scoring System (CCSS). Ten subjects voluntarily participated and completed the WAIS test. The best cutoff score of the CMT and the CDT using diagnosis of dementia as a gold standard were 15 and 7 respectively. Sensitivity and specificity for detecting dementia by the CDT were 100 per cent and 94.1 per cent respectively. Sensitivity and specificity for detecting dementia by the CMT were 83.3 per cent 91.7 per cent. The likelihood ratio (95% confidence interval) of the CMT and the CDT were 10 (3.3-30.4) and 17 (4.4-65.2) respectively. There was a good correlation of the CDT score and the CMT score with the WAIS score. Although the CDT was better than the CMT in literate patients, the CDT had limitation of use among illiterate patients but not the CMT. The benefits of simultaneously application on dementia screening was shown. This study showed that the CDT was a valuable, brief method of dementia screening in elderly Thai patients with memory problems. PMID- 10808692 TI - Imprint cytology from a childhood case of metastatic papillary thyroid carcinoma. AB - A 9-year-old girl with a history of a palpable multinodular hard mass in the right lobe of the thyroid gland was biopsied. On diagnosis of a papillary carcinoma, total thyroidectomy and right radical neck dissection were performed. Examination of frozen sections demonstrated metastasis in the right but not the left cervical lymph nodes. Imprint cytology revealed small papillary sheets of neoplastic cells with a high proportion of cytoplasmic inclusions and a few nuclear grooves. These nuclear details allowed a specific diagnosis of metastatic papillary thyroid carcinoma. Papillary thyroid carcinoma can be easily diagnosed by imprint cytology. In places such as small and country hospitals that do not have pathology laboratories, it can also be used successfully as an alternative to frozen section histology. The efficiency, simplicity and rapidity of this method make it a very useful procedure. PMID- 10808693 TI - Early complications of gastric transposition operation. AB - Gastric transposition was performed in 100 children as a definitive procedure for oesophageal replacement between 1982 and 1997 for 69 oesophageal atresia (41 with distal tracheooesophageal fistula, 20 isolated oesophageal atresia and 8 with proximal tracheooesophageal fistula), 16 severe caustic stricture, 7 intractable peptic reflux stricture and 8 miscellaneous causes. Six mortalities were recorded. Sixty-five patients had complications postoperatively and respiratory complication was the most common complication especially in oesophageal atresia patients. Swallowing difficulty, particularly in oesophageal atresia, occurred in 21 per cent of the patients. Ten patients developed cervical leakage with spontaneous closure and 8 patients suffered from anastomosis stricture. Six jejunostomy revisions were required. Three of five pyloromyotomy obtained inadequate gastric drainage post gastric transposition and required the conversion to pyloroplasty. Because of the distinctive low major life-threatening morbidity and low mortality, we concluded that gastric transposition was a safe, easy and preferable procedure for oesophageal replacement in children. PMID- 10808694 TI - Epidural analgesia for pain relief in thoracic surgery. AB - The effectiveness and adverse effects of continuous epidural analgesia was studied in 104 patients undergoing thoracic operations at Siriraj Hospital. Patients were divided into 3 groups according to the type of surgical approach and the technique of epidural analgesia. Group 1 patients (n = 72) received thoracic epidural block using bupivacaine and morphine combined with light general anesthesia for exploratory thoracotomy; group 2 patients (n = 21) received the identical anesthetic technique, the operation was achieved through median sternotomy; group 3 patients (n = 11) had a similar type of operation to group 1, the anesthetic technique was lumbar epidural block using morphine and combined with light general anesthesia. Continuous epidural morphine infusion was given 0.1-0.4 mg/h during postoperation in all patients for providing adequate pain relief. The results revealed that a 10 cm visual analogue scale (VAS) pain scores were satisfactory and comparable in all groups. Lumbar epidural patients consumed a significantly larger dose of morphine than thoracic epidural groups (P < 0.01). Intraoperative hypotension occurred 43.05 per cent and 19.05 per cent in group 1 and 2, but none was found in group 3 (P < 0.05). Postoperative respiratory depression was found 54.16 per cent in group 1, 33.33 per cent in group 2 and 9.09 per cent in group 3 (P < 0.05), and was mostly mild to moderate, except three patients in group 1 and one in group 2 who needed mechanical ventilatory support. There were no differences among the groups in the incidence of nausea/vomiting and pruritus. It is concluded that both thoracic and lumbar epidural morphine provide excellent postthoracotomy pain relief, whereas, respiratory depression is more common with thoracic than lumbar epidural morphine. PMID- 10808695 TI - Free disc fragment: a retrospective case analysis. AB - Preoperative diagnosis of free disc fragment is important in order to avoid treatment by chymopapain and percutaneous method of disc removal or anterior discectomy. This retrospective case analysis was to study the cause, physical findings and appropriate investigation to identify free disc fragment before surgery. One hundred and thirteen operative patients of herniated lumbar disc were studied. Of these, eleven patients (9.7%) were diagnosed with free disc fragment. The results revealed that one patient was diagnosed preoperatively with free disc fragment by myelography and MRI and three other patients using MRI alone. We concluded that preoperative diagnosis of free disc fragment can be made through identification of a large myelographic defect and/or sagittal scan MRI showing migration of disc material from native disc space. PMID- 10808696 TI - Uterine blood flow response to hormonal replacement therapy in asymptomatic postmenopausal women: a transvaginal Doppler study. AB - To evaluate the effect of continuous hormonal replacement therapy (HRT) on Doppler parameters of uterine blood flow in asymptomatic postmenopausal women. Thirty-eight asymptomatic postmenopausal women were recruited into the study from the outpatient menopause clinic, Department of Obstetrics and Gynaecology, Faculty of Medicine, Chulalongkorn University. The study population was divided into 20 cases without any HRT (group 1) and 18 cases using continuous conjugated equine estrogen 0.625 mg/day combined with medroxyprogesterone acetate 2.5 mg/day (group 2). The duration of HRT was 21.3 +/- 9.5 (13-56) months. A transvaginal colour flow imaging system (ALOKA SSD-2000 MultiView, Tokyo, Japan) was used to assess uterine blood flow. Quantitative data from areas of colour were evaluated by pulsed Doppler spectrum analysis. Resistance indices (RI) were measured as indicators of uterine perfusion. Both groups were statistically similar with respect to age, parity, age at menopause, height and weight. The endometrial thickness in group 1 and 2 were 3.8 +/- 0.8 and 4.1 +/- 0.6 millimetres, respectively. The left uterine artery RIs of group 1 and 2 were 0.86 +/- 0.08 and 0.84 +/- 0.07, respectively (p = 0.33). The right uterine artery RIs of group 1 and 2 were 0.87 +/- 0.07 and 0.83 +/- 0.06, respectively (p = 0.06). In conclusion, continuous HRT had a non-significant influence on uterine blood flow in the postmenopausal women. PMID- 10808697 TI - Amifostine and hematologic effects. AB - Amifostine is a protective agent of normal tissue from adverse effects of radiochemotherapy. It is the prodrug that is dephosphorylated by alkaline phosphatase on plasma membrane into the active form named WR-1065. More than 90 per cent of the drug is cleared from plasma in 6 minutes and the peak tissue concentration is 10-30 minutes after intravenous administration. Amifostine has the selective property to protect normal tissue but not cancer cells by mainly scavenging free radicals induced by radiation and chemocytotoxic agents. Both preclinical and clinical studies of this drug provide the significant protection of hematopoietic progentitors from a broad range of cytotoxic agents such as cyclophosphamide, cisplatin, vinblastine, carboplatin, mitomycin-C, fotemustine, doxorubicin, daunorubicin and radiation as well. Moreover, this drug can protect other normal organs or tissues including kidney, salivary gland, liver, heart, lung and small intestine. Amifostine is quite safe, the two major side effects are vomiting and hypotension, and the minor effects are flushing, sneezing, dizziness, chills, metallic taste etc. The drug was approved by the FDA of U.S.A. for use as a cytoprotectant in cyclophosphamide and cisplatin treatment for advanced ovarian cancer and non small cell lung cancer. PMID- 10808698 TI - Utility of serum cytokeratin 19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) as tumour markers for non-small cell lung cancer. AB - Serum cytokeratin 19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) levels were determined with an enzyme immunoassay in 51 patients with non-small cell lung cancer (NSCLC), 26 patients with benign lung diseases and 26 normal individuals in order to evaluate their clinical utility in the diagnosis of NSCLC. Patients with NSCLC demonstrated higher serum CYFRA 21-1 and CEA levels than both patients with benign lung diseases and normal group. We used the cut off value which was derived from the 95th percentile value of CYFRA 21-1 and CEA levels in the group of patients with benign lung diseases; CYFRA 21-1 at 3.13 ng/ml and CEA at 7.7 ng/ml. The sensitivity and diagnostic accuracy of CYFRA 21-1 and CEA for the group of NSCLC patients were 66.7 per cent, 76.6 per cent and 35.3 per cent, 55.8 per cent, respectively. When combining CYFRA 21-1 with CEA, the sensitivity and diagnostic accuracy were 68.6 per cent and 66 per cent. These results suggest that CYFRA 21-1 and CEA are useful serum markers for the diagnosis of NSCLC; especially subtype squamous cell and adenocarcinoma, respectively. The usefulness is not enhanced by combining the assay of CYFRA 21-1 and CEA. PMID- 10808699 TI - Nosocomial pneumonia in a newborn intensive care unit. AB - Nosocomial pneumonia is a major cause of morbidity and mortality in hospitalized patients. The risk is especially high in the neonatal intensive care unit (NICU) particularly in infants with mechanically assisted ventilation. During the 5-year period of the study, 160 infants with problems including prematurity (60.6%), respiratory distress (55.6%) and birth asphyxia (45.0%) were admitted to the NICU. One hundred and thirty-three infants (83.1%) received mechanical ventilation. Nosocomial pneumonia was found in 65 infants (40.6%) or 88.3 cases per 1,000 ventilator-days. Low birth weight, prematurity, respiratory distress and hyperbilirubinemia were found more significantly in the pneumonia group. They underwent more manipulations such as the placement of an umbilical catheter and orogastric tube. Infants with pneumonia received mechanical ventilation at a higher percentage and for a longer period than those without pneumonia (96.9% vs 73.7%, odds ratio = 11.2, p = 0.000) with a mean duration of 11.7 and 3.5 days respectively (p = 0.000). The etiologic organisms recovered from hemoculture were Acinetobacter calcoaceticus var. anitratus 44.0 per cent, Enterobacter spp. 16.0 per cent, Klebsiella pneumoniae 16.0 per cent, coagulase-negative staphylococci 12.0 per cent. There was no concordance of the bacteriologic results in endotracheal aspirate culture and hemoculture in each infant. Leukocytosis and granulocytosis as well as blood gas values could not differentiate the presence of pneumonia. The mean hospital stay for the infants with pneumonia was longer (23.0 days vs 6.4 days, p = 0.000). Nosocomial pneumonia did not only prolong hospital stay but also contributed to mortality. Twenty-seven (41.5%) of the infants with pneumonia died, compared with 46 (48.4%) of the other group without pneumonia (p = 0.422). The risk of nosocomial pneumonia can be reduced by using infection control measures, including meticulous hand washing and gloving during respiratory manipulation, heat-treated water supply in a nebulizing unit of the ventilator and proper care of umbilical catheterization. PMID- 10808700 TI - Lipiodol enhanced CT scanning of malignant hepatic tumors. AB - From August 1984 to March 1991, 41 patients with malignant liver tumors, 30 males and 11 females, aged 30-75 years were treated at Ramathibodi Hospital with injection of mitomycin-C lipiodol emulsion into the tumor via the feeding artery followed by embolization of the feeding artery with gelfoam particles. The patients comprised 30 cases of hepatocellular carcinoma, 4 cases of cholangiocarcinoma and 7 cases of metastatic tumors of which one was from CA stomach, three were from CA breast, and three from CA colon. The vascularity of the tumor was assessed in angiogram obtained prior to treatment and retention pattern of lipiodol in the tumor was evaluated in lipiodol-enhanced CT scan images taken 2-4 weeks following therapy. The results showed that lipiodol CT scan images exhibited four patterns of lipiodol retention in the tumor appearing as opacity as follows (1) homogenous (2) heterogeneous (3) ring-like and (4) none. Lipiodol retention pattern appeared to be somewhat related to vascularity of the tumor. Most of the hypervascular tumors such as hepatocellular carcinoma had homogeneous lipiodol accumulation pattern if the tumor size was less than 5 cm. Metastatic tumors and cholangiocarcinoma showed heterogeneous or ring-like pattern of lipiodol accumulation because they were relatively hypovascular. Hypervascular hepatocellular carcinoma may exhibit heterogeneous or ring-like pattern if they are larger than 5 cms, and have multiple feeding arteries, necrosis or AV shunting. Hepatocellular carcinoma with AV shunting may not show any lipiodol accumulation at all. PMID- 10808701 TI - Clinical effects of 17 beta-estradiol and norethisterone acetate in postmenopausal Thai women. AB - OBJECTIVE: To compare the climacteric symptoms, the bleeding patterns, side effects, endometrial thickness, serum follicular stimulating hormone (FSH) and estradiol level in postmenopause Thai women who received this drug and placebo. STUDY DESIGN: Double blind, randomized placebo controlled trial. MATERIAL AND METHOD: Sixty postmenopausal women attending the menopause clinic at Chulalongkorn Hospital from July, 1996 to December, 1996, were enrolled in the study. The patients were randomized to receive the placebo or drug (17 beta estradiol 2 mg and norethisterone acetate 1 mg) continuously. After 12 months, 13 patients in the drug group were switched to have placebo and 13 patients in the placebo were switched to the drug group. Recording of patient characteristics, physical and gynecologic examination, pap smear, breast examination and mammogram, climacteric symptom scores, transvaginal ultrasonography, serum FSH and Estradiol level were performed prior to the study. Physical examinations, breast palpitations, measurement of body weight and blood pressure, climacteric symptom scores and side effects were repeated at 3, 6, 12, 18 months. Gynecologic examination, pap smear, serum FSH and estradiol, transvaginal ultrasonography, were repeated at 12 months. RESULTS: The women in the drug group had fewer climacteric symptoms than baseline after 6 months. The incidence of amenorrhea was 74.0 per cent at 12 months. The incidence of abnormal uterine bleeding (spotting and breakthrough bleeding) was 37.0 per cent at 3 months, 29.6 per cent at 6 months, 25.9 per cent at 12 months and 7.1 per cent at 18 months. The women in the drug group had fewer climacteric symptoms than baseline after 6 months. Breast tenderness was the most common side effect. The endometrial thickness decreased significantly in the drug group. Serum FSH decreased and serum estradiol increased significantly in the drug group. CONCLUSION: This combined regimen was more effective in relieving the climacteric symptoms in women who used the drug than those who used the placebo. There was a high incidence of amennorhoea after 12 months. But there was also a high frequency of abnormal bleeding in the first 3 months of treatment. Counseling on the bleeding pattern and common side effects should be conducted before starting the treatment to increase the compliance. This combined regimen proved to be a useful alternative for postmenopausal Thai women who prefer to avoid cyclic bleeding from sequential therapy. PMID- 10808702 TI - Therapeutic effects of Aloe vera on cutaneous microcirculation and wound healing in second degree burn model in rats. AB - OBJECTIVE: To demonstrate the microcirculatory and wound healing effects of Aloe vera on induced second degree burn wounds in rats. METHOD: A total of 48 male Wistar rats were equally divided into 4 groups as follows: sham controls, untreated burn-wound rats, those treated with once-daily application of normal saline (NSS) and those treated with once-daily application of lyophilized Aloe vera gel. The animals in each group were equally subdivided into 2 subgroups for the study of cutaneous microcirculation and wound healing on day 7 and 14 after burn. Dorsal skinfold chamber preparation and intravital fluorescence microscopic technique were performed to examine dermal microvascular changes, including arteriolar diameter, postcapillary venular permeability and leukocyte adhesion on postcapillary venules. RESULTS: On day 7, the vasodilation and increased postcapillary venular permeability as encountered in the untreated burn were found to be reduced significantly (p < 0.05) in both the NSS- and Aloe vera treated groups, but to a greater extent in the latter. Leukocyte adhesion was not different among the untreated, NSS- and Aloe vera-treated groups. On day 14, vasoconstriction occurred after the wound had been left untreated. Only in the Aloe vera-treated groups, was arteriolar diameter increased up to normal condition and postcapillary venular permeability was not different from the sham controls. The amount of leukocyte adhesion was also less observed compared to the untreated and NSS- treated groups. Besides, the healing area of the Aloe vera treated wound was better than that of the untreated and NSS- treated groups during 7 and 14 days after burn. CONCLUSION: Aloe vera could exhibit the actions of both anti-inflammation and wound healing promotion when applied on a second degree burn wound. PMID- 10808703 TI - Scanning electron microscopic observations on advanced third-stage larva of Gnathostoma spinigerum after in vitro exposure to albendazole sulphoxide. AB - Gnathostomiasis is the parasitic disease caused by the migration of an advanced third-stage larva of Gnathostoma spinigerum. To date, albendazole is claimed to be the effective drug in preventing the reoccurrence of migratory swelling in patients. After being exposed to 1 and 2 micrograms/ml albendazole sulphoxide (AlbSO) in vitro, the parasites moved deteriorately, however, no dead larva was found even exposed to these concentrations for 21 consecutive days. The topographical alterations after 21 days of albendazole sulphoxide exposure are described using a scanning electron microscope. The marked changes in surface morphology were observed in both neck and body regions. The tegumental surface on the neck region was swollen and covered with fuzzy materials, whereas, the spines on the posterior region of the body were dislodged. These changes would probably lead to reduction of intermittent cutaneous migratory swelling in human gnathostomiasis patients. PMID- 10808704 TI - Pulse dye laser lithotripsy for large biliary tract stones. AB - From 1997 to 1998 the first series of 21 patients with large biliary tract stones (1.5-4.0 cm with average 2.5 cm in diameter) who underwent endoscopic stone extraction by using combination of ERCP and EST with automatic stone-tissue detection pulse dye laser was carried out. Male = 8, Female = 13, average age = 57.5 years (32-83) most of the stones were primary stones (71.4%). Laser fibers were controlled by balloon catheter with fluoroscope (blind technique) in 16 cases and by mother-babyscope system in 5 cases. The result in successful fragmentation of stones was 87.5 per cent with the blind technique and 100 per cent with the mother-babyscope technique. Temporary stents were used in the patients who had severe cholangitis and those who required more than one session of lithotripsy (19%). Complications consisted of controllable cholangitis in 2 cases (9.5%) and there was no mortality. We conclude that stone-tissue detection lithotripsy is very safe and effective in patients with large biliary tract stones and high surgical risk especially via the direct mother-babyscope system. PMID- 10808705 TI - Self-adhesive silicone gel sheet: a treatment for hypertrophic scars and keloids. AB - An open clinical trial was conducted to assess the effect of self-adhesive silicone gel sheet (SASGS) for the treatment of hypertrophic scars and keloids in Thai people. Patients were instructed to apply the SASGS to the scars as long as possible, but not less than 12 hours per day for at least 8 weeks. The subjective results of the treatment were evaluated by the patients. The scars were evaluated for color, height, weight before and after treatment at 4 and 8 weeks. Eighteen patients with 18 hypertrophic scars or keloids were recruited into the study. Their ages ranged from 6 to 33 years (mean 21 years). The average duration of the scars was 5.7 years. Twelve patients (66.67%) stated good results. All of the 18 patients wanted to continue the treatment with SASGS. Heights of the scars were reduced in 12 lesions (66.67%) after treatment for 8 weeks (P = 0.058). Weights of the lesions were decreased in 10 lesions (55.55%) but were not statistically different (P = 0.090). Seven lesions (36.84%) were improved in color. Two patients (11.11%) developed erythematous rash around the lesions which subsided after withdrawal of the treatment. The long term follow-up for the recurrence and the mechanism of action of this treatment should be studied further. PMID- 10808706 TI - Accuracy and precision of the i-STAT portable clinical analyzer: an analytical point of view. AB - The introduction of a new point-of-care testing (POCT) instrument into the market affects medical practice and laboratory services. The i-STAT is designed to improve the speed in the decision making of the medical profession. However, reliability of results would ensure the quality of laboratory data. We, therefore, made an evaluation of the performance of i-STAT using a disposable cartridge EG7 + which is capable of measuring pH, pO2, pCO2 (blood gas), Sodium, Potassium (Electrolytes), Ionized calcium and Hematocrit with only 10 microliters of lithium heparinized blood in 2 minutes. The results were compared with those obtained from routine methods. The results were found to be accurate, precise and correlated with acceptable methods used routinely in the laboratory. PMID- 10808707 TI - A case report: Alagille syndrome. AB - Alagille syndrome consists of 5 major features comprising paucity of interlobular bile ducts, characteristic facies, posterior embryotoxon, vertebral defects and peripheral pulmonic stenosis. The female patient in this report met 4 of the 5 major features except ocular abnormality. The first clinical presentations were prolonged jaundice and generalized ecchymoses. She was treated by plasma replacement and vitamin supplement, particularly vitamin K1, which produced clinical improvement. This report also reviews the literature of Alagille syndrome. PMID- 10808708 TI - [Morbidity of parasitic diseases in 1998 and measures for its stabilization in the Russian Federation]. PMID- 10808709 TI - [Regularity of formation of environmental parasitic contamination in urbanized ecosystems]. AB - The materials are summarized on the basis of which the authors were given Diploma No. 118 (with priority from November 23, 1994, registered under No. 137) for discovery of the same name by the resolution of the International Association of Authors of Scientific Discoveries on May 12, 1999. FORMULA OF THE DISCOVERY: "The unknown hitherto regularity of formation of environmental parasitic contamination in urbanized ecosystems has been ascertained to embrace the following. In emergency of biosocial conditions leading to parameters: (structures, functions, regulatory mechanisms) transformations of evolutionary established parasitic systems owing to the development of expression, expansion and succession in protozoa and multicellular parasitic organisms' populations, the conditions arise that favour the violation of biocenoses homeostasis and changes in tension of epidemic, epizootic and epiphytotic processes during parasitoses". The discovery pertains to the field of biology in particular, to general, ecological, medical, veterinary and agricultural aspects of parasitology. PMID- 10808710 TI - [Assessment of the association of morbidity due to parasitic diseases with the inoculation of the environment with their pathogens]. AB - The paper gives a conceptual design of field parasitological studies of the association of the inoculation of the environment with the pathogens of parasitic diseases with their spread among humans. The design includes 6 stages that reflect their object, the problems to be solved, guidelines, and expected results. It is of great importance in defining a factorial (environmental) risk for infection with the causative agents of parasitoses and in substantiating and implementing measures to protect the environment from pollution and to prevent parasitic diseases in man and animals. PMID- 10808711 TI - [Calculating the magnitude of the economic damage caused by diseases]. PMID- 10808713 TI - [Time course of serum nuclease activity in mice infected with Plasmodium berghei]. AB - The present paper shows that murine serum nuclease activity increases following P.berghei infection. DNA activity begins increasing just 48 hours after infection. Following 78 hours, it achieves the maximum, by exceeding the baseline level by 6 times. Then DNA activity starts decreasing and following 94 hours after infection it is just thrice higher than the baseline. Serum RNA activity shows only 30% increases 72 hours after infection and returns to the baseline following 94 hours. Microscopic monitoring indicates that only single malaria causative agents appear in the red blood cells in this period. The peak increases in nuclease activities after P.berghei infection are assumed to be associated with the induction of serum infection. PMID- 10808712 TI - [Investigation of the protein composition and immunochemical properties of Toxoplasma gondii excretory and secretory antigen]. AB - The major proteins of T. gondii excretory and secretory antigens (ESA) obtained during cultivation of tachyzoites by using cultured Vero cells were shown to have molecular weights of 79, 70, 57, 48, 36, and 29 kD. ESA and somatic antigen immunoblotting demonstrated that there were noticeable differences in the immunoactive proteins of these antigens. The antibodies of the sera of patients with toxoplasmosis mainly interacted with ESA proteins having molecular weights of 79, 70, 57, 48, 36, and 29 kD, 57-kD ESA protein antibodies being present on the immunoblots of all the tested sera. When ESA was used as an antigen during enzyme immunoassay, it showed a high sensitivity in the detection of IgM antigens in congenital toxoplasmosis. At the same time, ESA identified the antibodies of this class in the sera of healthy donors much infrequently than somatic antigen (p < 0.05). PMID- 10808714 TI - [Tanfelam: synthesis, study of ovicidal activity, and acute toxicity]. AB - The papers describes the synthesis of N-(2-piperidinoethyl)-N-tosyl-n-anisidine (Tanfelam) which showed a 100% ovicidal activity when tested by the Harada and Mori methods (in vitro inhibited N.brasiliensis hatching and development test). Tanfelam has been transferred for in-depth tests. PMID- 10808715 TI - [Comparative biopharmaceutical study of tegalide]. AB - The investigations have indicated a great influence of pharmaceutical factors on the pharmacokinetics of tegalide. The agent as tablets has a higher bioavailability than its substance. The high bioavailability of tegalide tablets appears to be associated with the presence of auxiliaries in them, which enhance the magnitude of the agent's absorption. Differences in the shelf lives of tablets affect their bioavailability. In this connection, the shelf life must be limited in terms of the pharmacological data presented. PMID- 10808716 TI - [Comparative efficiency of different formulations of insecticidal baits]. PMID- 10808717 TI - [Dynamics of the number of house dust mites and the accumulation of groups 1 and 2 allergens in simple periodic cultures]. AB - Dynamics of the number and biomass of D. pteronyssinus and D. farinae and the concentration of major significant mite allergens of groups 1 and 2 were studied in simple periodic cultures. The experiments provided evidence for that there is a positive relationship between the concentration of mite allergens Der p I, Der p II, Der f I, Der f II and the number of mites D. pteronyssinus and D. farinae, as evidenced by both correlation coefficients and curves. The same relationship was seen while examining the number of mites and the exposure of mite allergens in house dust. The findings suggest that the concentration of major mite allergens of Groups 1 and 2 in addition to the number/biomass of mites are one of the most important criteria for the maturity of the culture. The cultures of D. pteronyssinus and D. farinae mature no later than on day 29 of the onset of cultivation, as shown by the maximum concentrations of Der p I, Der p II, Der f I, Der f II and mite biomass, achieved in this period. PMID- 10808718 TI - [The disinfectant "Vibacide" for decontamination purposes in work with blockers]. AB - The authors propose the novel disinfectant Vibacide for decontamination of objects during experimental and experimental work against alive fleas. The disinfectant as 1-2% solutions had no toxic effect on Nosopsyllus consimilis fleas. PMID- 10808719 TI - [Effective scabies control system in Moscow]. AB - The analysis using the systems approach, analogue simulation, practical conclusions of the probability theory, mathematical statistical methods, programme-oriented work organization revealed and eliminated a number of neglects that had put bounds to scabies morbidity reduction among Moscovites. In 1998, the number of the foci that dermatovenereological facilities were unaware of substantially showed a 6.9-fold decrease. Therefore, the incidence of scabies reduced by 14.6% in the city in 1998. The paper presents the guidelines developed by the authors, which will ensure that the incidence rates of scabies be 25.07 per 100,000 Moscovites by 2005. It analyzes the scabies epidemiological situation among the homeless and possible measures to promote health in this group of persons. PMID- 10808720 TI - [Current epidemiology of parasitic diseases and tick-borne encephalitis in the Lvov region]. PMID- 10808721 TI - Variations in teenage birth rates, 1991-98: national and state trends. AB - OBJECTIVES: This report presents national birth rates for teenagers for 1991-98 and the percent change, 1991-98. State-specific teenage birth rates by age, race, and Hispanic origin for 1991 and 1998 and the percent change, 1991 to 1998, are also presented. METHODS: Tabular and graphical descriptions of the trends in teenage birth rates for the Nation and each State, by age group, race, and Hispanic origin of the mother, are discussed. RESULTS: Birth rates for teenagers 15-19 years declined nationally between 1991 and 1998 for all age and race and Hispanic origin populations, with the steepest declines recorded for black teenagers. State-specific rates fell significantly in all States for ages 15-19 years; declines ranged from 10 to 38 percent. In general, rates by State fell more for younger than for older teenagers, ranging by State from 10 to 46 percent for ages 15-17 years. Statistically significant reductions for older teenagers ranged from 3 to 39 percent. Reductions by State were largest for black teenagers 15-19 years, with rates falling 30 percent or more in 15 States. Among the factors accounting for these declines are decreased sexual activity, increases in condom use, and the adoption of the implant and injectable contraceptives. PMID- 10808722 TI - And now, a few words for the other side. PMID- 10808723 TI - Keep your divorce out of my dental office. PMID- 10808724 TI - Drug alert. PMID- 10808725 TI - Dental management of the patient with Alzheimer's disease. AB - This paper summarizes factors to be considered in the dental management of the patient who presents with Alzheimer's disease. The information contained in this article will expand the understanding of the etiology, suspected pathophysiology and the clinical features of patients with Alzheimer's who may present for oral rehabilitation. The objective is to better prepare the dental professional in order that he/she may provide effective treatment planning for these patients. PMID- 10808726 TI - Toxic epidermal necrolysis associated with quinidine administration. AB - Toxic epidermal necrolysis (TEN) is a rare, life threatening, drug induced cutaneous reaction first reported by Lyell in 1956. He named the condition TEN to distinguish it from staphylococcal scalded skin syndrome. It is characterized by a separation of the epidermis and dermis with subsequent desquamation of skin. The denuded areas of skin have the appearance of second-degree burn. Drug induced TEN is a disease of severe morbidity and high mortality. The drugs most frequently associated with onset of the condition are sulfonamides, anticonvulsants, non-steroidal anti-inflammatory drugs, and allopurinol. This study reports the occurrence of TEN in a patient receiving Quinidine. PMID- 10808727 TI - [Principles of evidence-based medicine; merits and pitfalls]. AB - Historical background, main principles, methodology of evidence based medicine (EBM) and prospective randomized trials (incl. megatrials) are reviewed. EBM has a significant beneficial influence on medical activities as follows: improving efficacy of medical therapy, generation of new research trends, optimizing decision making in clinical settings of incomplete pathophysiological background, exploring new associations can be found beyond the individual clinician's scope, regular financial support of gradual, postgradual medical training, medical research and international scientific programs. Potential adverse influences related to EBM are: weakening position of individual (versus modus) patient oriented approach in medical care, diminution of pathophysiology- (versus product ) oriented medical research, reevaluation of medical, scientific activity, interrelations between medical doctors and patients. Potential misuse of statistical methods in evaluation of megatrials is briefly discussed. A combination of benefits related to EBM and traditional elements of classical medical care, clinical research is needed to establish a more improved medical care for the individual patient. PMID- 10808728 TI - [Helicobacter infection in children]. AB - Consensus was achieved on the following issues: in children H. pylori infection causes chronic gastritis, but rarely gastric and duodenal ulcer disease. Eradication of H. pylori leads to healing of these conditions. To-date, there is no evidence demonstrating a link between H. pylori gastritis and abdominal pain except in those children where a gastric or duodenal ulcer is present. Children should be investigated for H. pylori infection only if their symptoms are suggestive of organic disease rather than functional abdominal pain. Endoscopy with biopsies is the optimal method to investigate a child with upper gastrointestinal symptoms suggestive of organic disease but this should only be carried out when a diagnostic work up using non-invasive methods has excluded other causes such as lactose maldigestion, constipation, coeliac disease, liver or biliary tact disease. If H. pylori is identified through investigations carried out during endoscopy the infection should be treated. Treatment should be monitored with a reliable non-invasive test and the 13C-urea breath test is the preferred method. H. pylori eradication in such children will cure gastritis but there is no data to support a relationship between a cure of H. pylori gastritis and symptom relief except in patients with ulcer disease. Further studies are needed to establish whether there are any specific symptoms associated with H. pylori gastritis alone and whether infected children without ulcer disease benefit from anti-H. pylori therapy regarding their symptoms. This consensus meeting did not deal with the optimal therapy for H. pylori infection as there are insufficient studies concerned the best treatment in children. PMID- 10808729 TI - [Tizanidine in the treatment of acute withdrawal symptoms in heroin dependent patients]. AB - One of the treatment alternative of withdrawal symptoms of patients suffering from opiate dependence is to apply the clonidine in combination or itself. This remedy is not in commercial trade in our country. It is expectable according to the recent data analysing the effects of the alfa2 adreneregic agonist tizanidine that tizanidine has the similar protective effect as clonidine with the resembling target point. Based on this theory a research was done, in the course of which the i.v. heroin users who presented themselves at the Drug Outpatient Department of Buda between 1.10.1998-8.01.1999. were divided into two groups. The groups had got the usual detoxification treatment, but in the experimental group tizanidine were given in 3 x 8 mg/day dose too. Sixteen patients were in the tizanidine group, 10 patients were in the control group. The patients estimated the intensity of 7 symptoms of withdrawal (sweating, nervousness, insomnia, tremor, diarrhoea, muscle pain, drug craving) on a subjective scale day by day. The analysis showed that the tizanidine treatment decreased the intensity of the withdrawal symptoms in every symptom type examined. The ten days long acute withdrawal period were accomplished by all of the patients, but in the short course of the following (mean 9 and 11 weeks in the treated and the control groups respectively) there were three relapses in each group (3/16 in the treated and 3/10 in the control). PMID- 10808731 TI - [A recovered case of actinomycosis in the small intestines diagnosed with ultrasonography]. AB - The authors report the history of a patient suffering from abdominal actinomycosis. The correct diagnosis could be established by an ultrasound-guided aspiration sampling from the small-intestinal abscess, following the sonographic localisation of the lesion. The patient recovered by a long-term antibiotic treatment. Related to the case the authors review the relevant literary data of the past few years and point to the value of ultrasonography in the successful management of the disease. PMID- 10808730 TI - [Detection of aneuploidy from gastrointestinal biopsy samples]. AB - Aim of the present work was the development of a mechanic cell separation protocol for gastrointestinal biopsy analysis. Evaluation of the technique was performed on selected group of patients who underwent routine endoscopy. Routine gastrointestinal biopsies were obtained after informed consent. 23 gastric (6 healthy, 14 gastritis, 3 adenocarcinoma) and 15 colon samples (5 healthy, 7 colitis ulcerosa, 3 adenocarcinoma) were evaluated. The mechanic disruption of the biopsies was performed by Medimachine (DAKO, Denmark), a commercially available system using a 30 microns miner and a 30 microns mesh. The cell solution was centrifuged for 5 minutes by 250 g. The cells were fixed in paraformaldehide and stained by propidium iodide. The flow cytometry analysis was performed on a BD FacStar Plus flow cytometer. The DNA data were evaluated using the Winlist software. All of the preparations were appropriate for flow cytometric analysis. The coefficient of variation of the DNA histograms (n = 7) (CV mean +/- SD. 6.45% +/- 1.21) were acceptable for analysis. In the gastric biopsy samples aneuploidy was determined only in malignant cases. In four of the seven colitis ulcerosa samples and in one of the three adenocarcinoma aneploidy was found. The histologically healthy specimen were all diploid. Mechanic cell separation and disaggregation is a useful method for preparing fresh biopsy specimen for flow cytometry. PMID- 10808732 TI - [Exemplary conduct of the teaching physicians is a big part of problem-oriented learning]. PMID- 10808733 TI - [Coronary artery disease in women]. AB - Data concerning coronary artery disease (CAD) refer to the recent clinical trials. They indicate that CAD is the most frequent cause of deaths of women over 50 years of age, and the dynamics of its development differs from that in male patients. The author discusses specificity of risk factors, effectiveness of various diagnostic methods, and different results of treatment of women with CAD (comparing to men). PMID- 10808734 TI - [Effects of nitrates and beta-blockers on platelet aggregation in patients with coronary heart disease]. AB - Our study aimed at evaluating an effect of the selected nitrates and beta adrenoceptor blockers on platelet aggregation in patients (pts) with coronary artery disease (CAD). The study included 168 male patients (M), aged between 33 and 72 years (mean age 51 +/- 7). 61 M given isosorbide dinitrate were divided into three groups: in the group I and II an effect of the drug on the platelet aggregation following a single 10 mg (I) and 20 mg (II) dose has been evaluated; in group III an effect of the drug after a two-week treatment has been evaluated. 14 male patients (group IV) were given 5-mononitrate 40 mg daily for two weeks. 85 male patients, treated with propranolol have also been divided into three groups. In group V and group VII an effect of propranolol on platelet aggregation following a single dose of 40 mg and 80 mg has been evaluated respectively. In the group VII an outcome of a two-week propranolol therapy has been assessed. Eleven patients (group VIII) received nadolol in a dose of 40-80 mg daily for two weeks. Platelet aggregation induced by adenosine diphosphate (ADP) in concentration of 1 mM/ml and 5 mM/ml (groups I, II, III, V, VI, VII) and only of 1 microM/ml (groups IV, VIII) was evaluated with Born's method. In patients with CAD only isosorbide dinitrate inhibits platelet aggregation. This effect has been noted following a single dose as well as a two-week treatment. Other drugs (5 mononitrate, propranolol, nadolol) transiently increase platelet aggregation, but became either ineffective after two-week therapy (5-mononitrate, nadolol) or increase (propranolol) platelet aggregation. Tendency to inhibit (sorbonit) or to increase (propranolol) platelet aggregation has been more pronounced with higher concentration of aggregating factor. PMID- 10808735 TI - [Idioventricular rhythm in patients with acute myocardial infarction: an indicator of perfusion or ischemic symptom]. AB - Episodes of idioventricular rhythm occur frequently during the first hours of myocardial infarction. In patients receiving thrombolytic therapy, idioventricular rhythms may be an indicator of successful reperfusion--called "reperfusion arrhythmia". The aim of our study was to assess an incidence of idioventricular rhythm and other ventricular arrhythmias and their relations to thrombolytic therapy and reperfusion in a relatively large group of patients with acute myocardial infarction (AMI). The study included 100 patients with AMI, divided into three groups: with anterior, inferior and inferior with right ventricle infarction. Sixty six patients were treated with thrombolysis. 24-hour Holter recordings were obtained in every patient and reperfusion was assessed clinically. Episodes of idioventricular rhythm and other ventricular arrhythmias were more frequent in the thrombolytic group but not in patients with clinically established reperfusion. The main conclusion from our study is, that accelerated idioventricular rhythm is not specific for reperfusion but rather for myocardial ischaemia. The higher incidence of the ventricular arrhythmias in the thrombolytic group partially depends of their earlier hospitalization, and time dependent inclusion into treated group. PMID- 10808736 TI - [The QT interval dispersion and ventricular late potential in obese women]. AB - Obesity is known to be a risk factor of cardiac death, that is associated first of all with cardiac arrhythmias. Increased QT dispersion (QTd) and ventricular late potentials (LP) are measurable indices of ventricular arrhythmias risk. The aim of this study is to asses QTd and LP in women with obesity. 62 obese women (mean BMI 36.7 kg/m2) and 15 apparently healthy subjects (mean BMI 24.5 kg/m2) were included in our study. QTd and correlated QT interval dispersion (QTdc) were calculated from 12-lead ECG. LVM were assessed from echocardiograms. LP were obtained by signal averaging of surface electrocardiograms. RESULTS: QTd, QTdc, LVM were significantly higher in patients. We found LP in six cases and higher QTdc in this subgroup. According to our research, increased QTd in obese women seems to be associated with left ventricular hypertrophy and increased QTd is significantly higher in subjects with LP. PMID- 10808737 TI - [ECHO image quality of the left ventricle during dobutamine stress echocardiography]. AB - This study aimed at assessing ECHO image quality of the left ventricle during dobutamine stress echocardiography (ED). The study involved 582 patients without previous myocardial infarction. Their age ranged from 27 to 74 years, mean age 52 +/- 9. Dobutamine was given in stepwise increasing doses from 5 to 40 mcg/kg per minute. Atropine was added in 253 (43%) cases. The wall motion asynergy was evaluated by left ventricular echo image divided into 16 segments. At least one segment has not been visualised in 5.5% of patients. At the peak dose of administrated dobutamine (> 20 mcg/kg per minute) it increased to 6.1%. Unvisualized segments did not impair the test and did not affect negative or non diagnostic results, as was verified statistically. Patients' age, body weight and BMI did not exert a significant effect on the quality of echo image. Segments 5 and 11 were seen constantly during the whole test. Segment 13 was poorly visualised during the tests. Only in 1.5% of patients peak dose of dobutamine deteriorated the image, especially in segments 1, 3, 6, and 12. During the dobutamine echocardiography test the quality of ECHO image of left ventricular wall was worsening only in a few patients. It was not considered the reason of test interruption and did not impared the results of ED. PMID- 10808738 TI - [Quality of life two years after myocardial infarction. 1. Examination questionnaires]. AB - The purpose of the present study was to assess the quality of life (QOL) and factors influencing QOL in patients 2 years after myocardial infarction (MI). The study was performed in 100 pts (mean age 45.5) mean 2 years after MI. 50 pts of studied group (mean age 42.5) were resuscitated from ventricular fibrillation (VF) complicating acute phase of MI. The control group consisted of 100 healthy subjects persons (mean age 53.5). The QOL index was assessed with the use of 2 types questionnaires: "Quality of Well-Being Scale" (QWB) and questionnaires of QOL including 26 factors in terms of: pain, dyspnoea, anxiety, depression, energy, sleep, emotions, mobility, social isolation, sexual activity, hobby, subjective emotional, distress, functional and social state and return to work. The QOL index was assessed using mathematical Krefft method. The average value of QOL index in patients 2 years following MI was significantly lower when compared with control group. The mathematical method of Krefft showed, that out of 26 features the most important diagnostic power factor for the QOL index were: self estimation of physical performance, return to work, the lifestyle and presence of stenocardial pain and dyspnoea. The pts resuscitated from VF revealed the higher level of anxiety. PMID- 10808739 TI - [Quality of life two years after myocardial infarction. 2. Examination of affecting factors]. AB - This study aimed at determining these factors--ventricular fibrillation (VF) in acute phase of myocardial infarction MI), thrombolytic therapy, beta-blockers after MI, cardiac rehabilitation--affect quality of life (QOL) during the follow up. The study involved 100 patients (mean age 45.5 years), on the average 2 years after MI, MI has complicated VF in 50 patients. Questionnaire of QOL after MI developed internally, taking into account 26 features (clinical, emotional and social status), and estimating anxiety level (STAI) were used. Computer medical diagnostics "SyntMED" (Krefft's method) was used. In both groups after MI (complicated and not complicated VF) beneficial effect of: thrombolytic treatments in acute phase MI (0.65 vs conventional 0.45, p > 0.05), use of beta blockers after MI (0.56 vs 0.41 p < 0.05), and cardiac rehabilitation (0.64 vs 0.44 p < 0.05) on value QOL were noted. Patients after MI complicated VF had higher level of anxiety. Patients who returned to work had higher index QOL (0.62 vs 0.39). Ventricular fibrillation complicated the acute phase of MI had no significant effect on follow-up QL. PMID- 10808740 TI - [An influence of cardiovascular risk factors and pharmacotherapy on the long-term results in women undergoing coronary artery bypass]. AB - This study aimed at analysing an effect of the coronary risk factors and pharmacotherapy on the long-term outcome in women following the coronary artery by-pass. In 1004-1997, 253 female patients, aged between 33 and 82 years (mean [+/- SD] 57.0 +/- 8 years) were treated surgically. The follow-up period lasted for 7 to 60 months (mean 32.0 +/- 14 months). Ten patients (3.9%) died. Answers to the questionnaire and personal interviews assessed physical fitness based on CCS classification, pharmacotherapy, and presence of risk factors. According to CCS scale, significant improvement has been seen in 195 (82.6%; p < .0001) patients. Health state did not change in 34 (14.4%) patients, and deteriorated in 7 (3.0%). Analysisn coronary risk factors, hypertension proved prevailing (60.3%), followed by diabetes mellitus (25.5%) and obesity (22.9%). Eleven percent of patients returned to cigarettes smoking after surgery. Postoperatively, 74.1% of patients received nitrates as a constant, medication, 58.2%--beta-blockers, 53.4%--ACE inhibitors, and 19.8% of patients received calcium antagonists. Lipid abnormalities have been treated in 49.1% of patients whereas antiplatelet therapy has been carried out in 74.1%. Only 9.9% of patients received hormones. The lower CCS class before surgery, the more significant improvement after it. As pharmacotherapy was used according to the European guidelines, an improvement in the long-term outcome required some modifications in patients' life style. PMID- 10808742 TI - [Cardiovascular disease prevention in the elderly]. AB - In Poland and all over the world, there is a significant increase in the number of elderly persons (over 65 years). Evidently, cardiovascular disorders occur more frequently in the middle-aged and older people. Approximately 60 to 75% of persons over 65 years and older suffer from the myocardial ischaemic disorder or atherosclerosis. In the elderly, the same risk factors as in the other age groups are present, i.e. lipids metabolism failure, arterial hypertension, tobacco smoking, carbohydrates metabolism dysfunction, obesity, the lack of physical activity. The right arterial hypertension and lipids and carbohydrates metabolism dysfunction treatment connected with the cut out of the tobacco smoking are highly beneficial in the elderly suffering from the myocardial ischaemic disorder. PMID- 10808741 TI - [Pertrochanteric fractures of the femur in patient with thrombophlebic crural ulcerations of the legs in the course of 50 years]. AB - The authors present a case of 75-year old female, who has suffered from venous insufficiency crural ulcerations and lost hope for any relief for 50 years. Hospitalisation for a femur fracture has been a turning point. The patient learnt that modern medicine can treat disease that she suffered from for most of her life. PMID- 10808743 TI - [Leptin and obesity]. AB - Obesity is associated with increased incidence of cardiovascular diseases, insulin resistance, dyslipoproteinemia and cancer. The discovery of leptin in 1994 has provided a lot of new information about obesity. Leptin is a 167-amino acid peptide synthetized almost exclusively in adipose tissue. This hormone circulates in blood serum in both free and bound forms. The long isoform of leptin receptor is widely distributed in brain, whereas numerous short forms are also being present in peripheral tissues. Leptin acts by binding to the receptors in hypothalamus and altering a release of several neuropeptides, especially neuropeptide Y, regulating energy intake and expenditure. Apart from signaling energy reserves to the brain, leptin promotes hematopoiesis, influences pubertal development and contributes to the increase in arterial blood pressure. Leptin production regulation in humans is poorly understood, but appears to depend on the total body fat, changes in energy intake and serum level of several hormones. Despite the recent advances in the knowledge of both physiology and pathophysiology of leptin, several many important questions require further studies. PMID- 10808744 TI - [Recurrent aphthous ulcers: the etiology with special reference to immunological theories]. AB - The etiopathology of recurrent aphthous ulcers (RAU) is still unknown. Many local and systemic factors can participate in pathogenesis of recurrent aphthous ulcers. The authors discuss major theories of RAU etiology and especially concentrate on the role of immune system disorders. PMID- 10808745 TI - [Dressler's syndrome]. PMID- 10808746 TI - [Environmental health: concepts and activities]. AB - The objective of this study is to provide information and a conceptual framework that will facilitate the work of persons in charge of systematizing institutions devoted to environmental health. The notion of "environment" is examined and a definition is proposed, while a look is also taken at the place held by environmental health within the context of environmental problems and their "green" and "blue" components. A number of definitions are put forth before presenting the official definition of environmental health issued by WHO in Sofia (1993). Subsequently a list is presented of the basic areas that have been assigned to the field of environmental health by different organizations or at various meetings, with PAHO, WHO, and Program 21 among them. From this a rather exhaustive list of areas and subareas is constructed, with the finding that all lists are, in reality, an asystematic grouping of three different components: determining factors (from the physical world), processes (types of interventions), and roles (series of administrative tasks) which can be conceived as a matrix and which allow for the description of individual activities by the environmental health services. Certain rules of operation are proposed which make it possible, through a form of algebra, to construct expressions for describing such activities and their aggregates with some precision. Thus, it becomes possible to employ a common symbolic language which can facilitate intercommunication, teaching, and research in the area of environmental health. PMID- 10808747 TI - Attitudes towards mental illness in the Commonwealth of Dominica. AB - Little is known about the perception of mental illness in the English-speaking Caribbean. This study was conducted in 1995 to determine the attitudes, knowledge, and help-seeking practices for emotional disorders in the Commonwealth of Dominica. Two groups in Dominica were surveyed: 67 community leaders, consisting of nurses, teachers, and police officers; and 135 community members grouped into five socioeconomic strata that were collapsed to three for the analysis. All the respondents were asked to identify and suggest management of individuals with psychosis, alcoholism, depression, and childhood hyperactivity, as depicted in case vignettes. The person in the psychosis vignette was diagnosed as suffering from mental illness by 84.0% of the leaders and by 71.2% of the community members. However, in each of the three other vignettes, fewer than 30% of the respondents thought that mental illness was present. The person with alcoholism was viewed as having a serious problem by only slightly more than half of the respondents. Fewer than half of the respondents thought that the individuals with depression or hyperactivity had serious problems. The community leaders did somewhat worse in recognizing mental illness than did the community members. Respondents were most likely to refer a family member with emotional problems to a medical practitioner. In conclusion, education about mental health problems is needed in Dominica. Especially disconcerting was the lack of knowledge on mental illness among nurses, teachers, and police officers, that is, professionals directly involved in the pathway to care. PMID- 10808748 TI - [Mass production of Romanomermis iyengari (Nematoda: Mermithidae) applied to anopheline breeding sites in Boa Vista (Roraima), Brazil]. AB - In order to begin mass producing the nematode Romanomermis iyengari Welch, a bioprocessing plant was designed and set up on the grounds of Universidad Federal in the State of Roraima, Brazil, after reaching an agreement with the State's Health Department. The objective of this paper was to establish the basic process for mass breeding the parasite in order to apply it to anopheline breeding sites. Cultures were obtained at a rate of 68 during every seven-day cycle, making for a total of 272 cultures a month. Before treatments were applied in the field, laboratory tests were conducted that showed the great susceptibility of anopheline larvae to infestation by R. iyengari, with parasitism rates ranging from 71 to 98%. In order to assess the parasitizing capacity of R. iyengari in actual field conditions, 12 natural anopheline breeding sites were chosen, each ranging in size between 50 and 450 m2. The species Anopheles albitarsis Lynch Arribalzaga and Anopheles rondoni Neiva-Pinto were detected there, at densities that ranged from 34 to 66 larvae/m2. The biolarvicide was sprayed in all 12 breeding sites with a domestically manufactured pump set at a pressure of two atmospheres and a dose of 2,000 preparasites/m2. Seven days after treatments were performed, anopheline populations were markedly reduced (85 to 97%). Results obtained point to the feasibility of using R. iyengari to control larval populations of both species of anophelines. PMID- 10808749 TI - Malaria in the Limbe River valley of northern Haiti: a hospital-based retrospective study, 1975-1997. AB - In the Limbe River valley of northern Haiti a retrospective study at the Bon Samaritain Hospital (BSH) determined the total number of cases and the cyclical nature of malaria from 1975 through 1997, examined the relationship between rainfall and malaria from 1975 through 1985, and compared the incidence of malaria at that hospital with general trends for Haiti for 1975 through 1996 as reported by the World Health Organization (WHO). During 1975-1997, 27,078 positive cases of malaria were diagnosed at BSH; 50% of these cases occurred during 16 weeks out of the year, during a summer peak in June and July and a winter peak in December and January. For 1975-1985, there was no significant correlation between the incidence of malaria and annual rainfall. The strongest correlation was observed between weekly rainfall and weekly incidence of malaria when the data was staggered to allow a lag of 9-11 weeks between rainfall and new malaria cases. The lag period is explained by the time required for the creation of breeding sites after rain, the life cycles of the Anopheles albimanus mosquito and the Plasmodium falciparum parasite, and the incubation period for falciparum malaria. The incidence of malaria in the Limbe River valley loosely followed the trends in all of Haiti and also supported WHO reports indicating that malaria in Haiti has been in a general decline since the mid-1980s. By showing the seasonal trends for malaria in the Limbe valley and the relationship between rainfall and malaria over an extended time period, this study provides a means to measure the effectiveness of malaria control efforts in the region. PMID- 10808750 TI - [Abortion-related mortality in Brazil: decrease in spatial inequality]. AB - Abortion is not only a major cause of obstetric hospitalization in poor countries, but it also represents the failure of the public health system to provide enough information about contraceptive methods and thus prevent pregnancies. In Brazil, the high utilization rates of health facilities due to abortions reflect the ongoing difficulties with family planning and contraception. In addition, mortality resulting from abortions serves as an indicator of the quality of abortion procedures, an important point in a country where the practice is illegal and therefore done clandestinely. In this study, we analyzed the rates of mortality resulting from abortions among women 10 to 54 years old, including women who died from spontaneous and induced abortion, from 1980 to 1995, for the various regions of the country. The information we used came from the mortality data bank of the public health system of the Ministry of Health. Population data were obtained from the Brazilian Institute for Geography and Statistics. We studied 2,602 deaths, 15% of which were due to missed abortion, spontaneous abortion, or legally permitted induced abortion. The other 85% of the deaths were due to illegal induced abortions or to nonspecified abortions. The mortality rates from abortion-related causes have steadily decreased in all the regions of Brazil, but this improvement has been unevenly distributed in the country. The region with the smallest decrease in this rate (38% over 15 years) was the Northeast. The age of women dying from abortions progressively declined over the period studied. PMID- 10808751 TI - [Knowledge about Chagas' disease and risk factors in Argentina communities with different epidemiological trends]. AB - Currently, Chagas' disease control consists mainly of chemical warfare against the insect vector. However, a number of risk factors, such as poor hygiene and clutter, can facilitate the persistence of triatomine breeding sites. Relying on the premise that communities at risk of Chagas have little knowledge about the disease, the authors defined the extent of knowledge that is considered optimal and determined how much is known, on average, by the inhabitants of two areas in Argentina showing different epidemiologic trends. Risk factors in both areas were identified. The optimal extent of knowledge was defined in accordance with 25 "basic concepts" surrounding the disease, and from these questionnaires were constructed for evaluating average knowledge about the disease. Results obtained showed that risk factors were linked primarily with the type of dwelling construction and with clutter, as well as with limited knowledge about the disease. There was little basic knowledge about Chagas, with an inability, for example, to identify the vector and describe the mode of transmission. Increasing people's knowledge about the disease would be an important step in the fight against Chagas and would give the inhabitants of endemic areas a better understanding of their situation as well as the chance to develop behaviors that would allow them to assume responsibility for their own welfare. PMID- 10808752 TI - A bacteriological study of hospital beds before and after disinfection with phenolic disinfectant. AB - In hospitals, one of the ways to control microbial contamination is by disinfecting the furniture used by patients. This study's main objective was to evaluate the microbiological condition of hospital mattresses before and after such disinfection, in order to identify bacteria that are epidemiologically important in nosocomial infection, such as Staphylococcus aureus and Pseudomonas aeruginosa. RODAC plates with two different culture media were used to collect specimens. Patient beds were selected according to previously established criteria, and surface areas on the mattresses were chosen at random. From the total of 1,040 plate cultures from 52 mattresses, positive results were obtained from 500 of them (48.1%), 263 before disinfection and 237 after disinfection. Considering the selectivity of the culture media, the positivity rate was high. There were high prevalences of S. aureus both before and after mattress disinfection. The study results suggest that the usual disinfection procedures, instead of diminishing the number of microbes, merely displace them from one part of the mattress to another, and the number of microorganisms remains the same. PMID- 10808753 TI - Technology evaluation of a USA-Mexico health information system for epidemiological surveillance of Mexican migrant workers. AB - From 1994 through 1996, federal, state, and nongovernmental organizations in Mexico and in the United States of America developed and piloted a Binational Health Information System for Epidemiological Surveillance of Mexican migrant workers. The system allowed data exchange for epidemiological surveillance between the state of Guanajuato in Mexico and the Commonwealth (state) of Pennsylvania in the United States, for case detection, prevention, and treatment, through shared contact investigation and case management of communicable diseases. The target population consisted of migrant workers traveling between Guanajuato and Pennsylvania to work mainly in the mushroom industry, and their sexual partners in their Mexican communities of origin. Computerized migrant health information modules were set up in Guanajuato and in Pennsylvania. Patient information and epidemiological surveillance data were encrypted and communicated electronically between the modules, using the WONDER communications system of the U.S. Centers for Disease Control and Prevention. Evaluation of the Guanajuato Pennsylvania Binational Health Information System showed that major barriers to binational epidemiological surveillance and control are: a) lack of communication binationally; b) interrupted medical care due to migration; c) inconsistent diagnosis and treatment criteria between the two countries; d) lack of referral clinical records from one country to the other; and e) deficient legal regulations concerning binational clinical data transfer. To our knowledge, this is the first project that has successfully demonstrated the technological feasibility of a binational disease control system linking a state in the interior of one country with a state in the interior of another country, rather than just states in the border region. The project also advanced the understanding of health service organizational issues that facilitate or hinder communication, outreach, disease prevention, and organization of health care services for migrant workers in both Mexico and the United States. Despite the unprecedented success and potential bilateral benefits demonstrated by this project, serious structural and organizational deficits in the public health systems of both countries must be addressed before epidemiological surveillance can be achieved binationally. PMID- 10808754 TI - [Some observations on taxonomic nomenclature]. PMID- 10808755 TI - The National Nursing Home Survey: 1995 summary. AB - OBJECTIVE: The 1995 National Nursing Home Survey (NNHS) was conducted to collect data on nursing homes and their current residents. This report presents detailed data on the characteristics of the nursing homes including ownership, certification, bed size, location, affiliation, and services provided. Data on current residents are presented by basic demographics, living arrangement prior to admission, functional status, and other health and personal characteristics of the residents. METHODS: The 1995 NNHS is a sample survey consisting of a two stage design with a probability sample of 1,500 nursing facilities in the first stage and up to six current residents from each facility in the second stage. RESULTS: About 1.5 million residents were receiving care in an estimated 16,700 nursing homes in 1995. Nearly 1.8 million beds were available and facilities operated at about 87 percent of their capacity. Nearly 90 percent of the residents were 65 years and over. They were predominantly female and white with a large portion needing assistance in the activities of daily living (ADL's) and instrumental activities of daily living (IADL's). PMID- 10808756 TI - A good night out on the town. PMID- 10808757 TI - Diabetic emergencies: Part 1. Hypoglycaemia. AB - Diabetes mellitus is a chronic, lifelong condition which can affect people of all ages, and is increasing in prevalence. Hypoglycaemia is probably the most common acute problem suffered by patients with diabetes. It is also a serious medical emergency with potentially fatal outcomes, and is the most common reason for patients with diabetes attending an accident and emergency (A&E) department. It is also a major source of anxiety for diabetics, particularly those controlled on insulin, and unfortunately, in the move towards ever tighter glycaemic control, it is inevitable that diabetics will continue to suffer from hypoglycaemia. This article examines the pathophysiology of hypoglycaemia and some of its main causes, and will look at the clinical management of the patient with hypoglycaemia, both in the community and in the A&E department. The importance of the recognition and prompt treatment of hypoglycaemia, and of the investigation of hypoglycaemia with no obvious cause are also discussed. Part 2 of the series will explore the pathophysiology and clinical management of diabetic emergencies involving hyperglycaemia, including both diabetic ketoacidosis and the rarer hypernatremic, non-ketotic coma. PMID- 10808758 TI - Taking care of your feet. AB - For anyone who finds walking and standing painful, the working day can seem interminable and miserable. This is compounded for nurses who lead busy, active lives, and foot problems can become a major barrier to an effective working day. Chronic foot problems have led nurses to seek alternative employment, often outside the profession, thus leaving health services even less well staffed with experienced personnel. Foot health, then, plays an important role in maintaining the mobility of nurses and foot care is vital for continued comfort during gait. It is equally important to understand which foot problems may be safely dealth with by the sufferer and to identify those that require specialist help and advice from a State Registered Chiropodist/Podiatrist. PMID- 10808760 TI - NHS Direct. Interview by Lynn Sbaih. PMID- 10808759 TI - Comparison of current local practice and the Ottawa Ankle Rules to determine the need for radiography in acute ankle injury. AB - Ankle injuries are a frequent cause of attendance to accident and emergency (A&E) departments. Although often classed as 'minor injuries', the consequences of mismanagement can be debilitating for patients. In addition, it is not always easy to differentiate those injuries with bony fractures from solely ligamentous injuries, and most patients are radiographed unnecessarily. Clinical prediction rules such as the Ottawa Ankle Rules have been devised to reduce the degree of radiographic requests, thus limiting harmful radiation exposure, and reducing departmental costs. The A&E environment of our centre often includes a variety of clinicians with little pure orthopaedic training or knowledge of these clinical prediction rules, hence, providing a prime atmosphere for their assessing their effectiveness. A prospective study was performed in our centre to investigate the validity of the clinician's decision in diagnosis of a bony injury in the absence of training with the Ottawa Ankle Rules. The effectiveness of applying the rules was then determined on the same set of patients. PMID- 10808761 TI - Living forensics: a natural evolution in emergency care. AB - 'Throughout history, health care professionals have been called upon to assist the legal system in the prosecution of cases where patient care overlaps with the law and physiological realities collide' (Lynch 1995). Working for the last 21 years in the Accident & Emergency setting, the author is dedicated to increasing the awareness and assisting in the establishment of basic forensic evidence collection guidelines for the emergency care provider. Due to the nature of the clientele and setting, emergency personnel will inevitably care for 'victims of violence'. Domestic violence injuries, abuse and neglect in the elderly and young, the addictive client seeking emergency care, the sexual assault victim, and sufferers of occupational injuries, are but a few of the cases that would be classified in the forensic arena. Holistic care dictates looking after the patient as a whole. The nurse must meet the patient's physical and psychosocial needs. The forensic nurse ensures that the patient's civil and constitutional rights are also met. This forensic health care role can be achieved and strengthened by recognizing potential evidence and maintaining a 'chain of custody' of this evidence. PMID- 10808762 TI - Reasons why patients bypass their GP to visit a hospital emergency department. AB - Knowledge about what motivates patients to visit the emergency department (ED) of a hospital for minor complaints, instead of visiting their general practitioner (GP), can help to reduce unnecessary utilization of expensive services. This paper reports on a study designed to investigate the reasons why patients visit the ED and to determine the influence of patient characteristics on specific motives. A multidimensional measurement instrument was designed to identify the motives of patients who bypass their GP and visit the ED. The instrument assessed 21 motives, all measured by means of three questions in Likert format. During a period of 1 week, all patients who visited the ED of two hospitals in Amsterdam were asked to complete a questionnaire when they were 'self-referred' with minor complaints. A total of 403 questionnaires were analysed, and the results show that motives relating to the GP play a minor role in the decision of patients to visit the ED. Profiles of two major patient groups could be identified. One group comprised patients with a high socio-economic status living in suburbs, whose motives for visiting the ED are mainly of a financial nature. Patients in the second group mainly lived in the inner-city, and preferred the expertise and facilities provided by the ED. PMID- 10808763 TI - Techniques to comfort children during stressful procedures. AB - Medical procedures can be unpleasant experiences for children, their parents, and health care providers. We present this model of working with children having invasive procedures with the aim of helping to increase the comfort of infants and children and also parents and medical staff. The model has five parts: (1) Preparing the child and parent for the procedure and for their role during the procedure; (2) inviting the parent/caregiver to be present; (3) utilizing the treatment room for stressful procedures; (4) positioning the child in a comforting manner; and (5) maintaining a calm, positive atmosphere. PMID- 10808764 TI - Dilemma. Prisoners and hospitals. PMID- 10808765 TI - The power of the pen. PMID- 10808766 TI - Telephone triage: good business. PMID- 10808767 TI - Helping a dad quit smoking. PMID- 10808768 TI - Exams reflect NP role. PMID- 10808769 TI - Prescribing trends in the treatment of acute otitis media. Reining in resistant bacteria. AB - Several factors influence antimicrobial selection when treating otitis media. In addition to good in vitro activity against common pathogens, the drug should be well-absorbed with high serum concentrations and good penetration to the site of infection. Oral amoxicillin dosed at 40 mg/kg/day to 50 mg/kg/day every 8 hours for 10 days remains first-line therapy for uncomplicated AOM. Second-line agents to treat AOM can include second- and third-generation cephalosporins. The rapid emergence of antibiotic-resistant bacteria represents a particular problem because these pathogens cause not only otitis media but more serious and invasive bacterial infections such as pneumonia, bacteremia and meningitis. PMID- 10808770 TI - Appetite suppressants and cardiac valvulopathy. Current clinical perspectives. AB - Primary pulmonary hypertension (PPH) in association with weight loss treatment was initially observed in the late 1960s and early 1970s with the diet pill aminorex fumarate. Recent data from large-scale studies and single sites have found prevalence rates of anorexic-associated valvulopathy ranging from 15% to 30%. But overall, no increased prevalence of VHD has been observed among case control groups. In March 1998, the American Heart Association and American College of Cardiology released guidelines for the management of valvular heart disease. A section of the report was dedicated to patients exposed to anorectic drugs, and those statements are outlined in this article. PMID- 10808771 TI - An emerging epidemic. Chronic hepatitis C in children. PMID- 10808772 TI - Kid stuff. Teaching children about diabetes management. PMID- 10808773 TI - Into the woods. New weapons against Lyme disease. PMID- 10808774 TI - Advances in the diagnosis and treatment of fecal incontinence. Biofeedback in the forefront. PMID- 10808775 TI - Establishing an electronic presence. How to build your own web page. PMID- 10808776 TI - Continuing education on the Internet. Online options for NPs. PMID- 10808778 TI - Urgent care. Treating sexual assault victims. PMID- 10808777 TI - Dermatologic issues in the elderly. Examining common problems. PMID- 10808779 TI - Synagis protects high-risk infants against RSV and LRTI. PMID- 10808780 TI - The eye of the hurricane. PMID- 10808781 TI - Reporting child maltreatment: ethical issues for the nursing profession. PMID- 10808782 TI - The specialist breast care nurse: an evolving role. AB - The role of the specialist breast care nurse has not previously been described in the Australian context. A study was undertaken, utilising the Delphi technique and focus group interviews to determine the key elements of the role and to describe how these role elements are enacted by the nurses incumbent in these positions in six Australian states. Three rounds of questionnaires to 16 expert nurses established a consensus view as to the essential elements of the specialist breast care nurse's role. The 11 role elements that became significant during analysis of these data were: specialist nurse, supporter, educator, counsellor, adviser, team member, resource person, caregiver, public advocate, manager and researcher. Focus groups were held with nurses from each state and the findings allowed enhancement of the data from the questionnaires. Analysis of the interview data allowed a fuller description of how the nurses actualised their role. This data showed that the specialist breast care nurse co-ordinated the care of the woman and her family during the breast cancer journey. Integral to the role was the support for the woman and her family. It is concluded that in order to perform successfully in the role nurses require specialist education in supportive care, counselling, pathology and treatment of breast cancer, a broad knowledge of oncology nursing, management, research and teaching techniques. PMID- 10808783 TI - Refractive surgery. AB - This article presents an overview of the terms used in laser refractive surgery and identifies the benefits, outcomes, complications and shortcomings of these procedures. Even though it is 'fashionable' to have these procedures done, they are still largely experimental. Although there are benefits from these procedures, thorough screening and selection of all patients should be performed. PMID- 10808784 TI - Effects of size of health service on scope of rural nursing practice. AB - This paper presents the findings of an analysis of the activities of rural nurses from a national audit of the role and function of the rural nurse (Hegney, Pearson and McCarthy 1997). The results suggest that the size of the health service (defined by the number of acute beds) influences the activities of rural nurses. Further, the study reports on the differences of the context of practice between different size rural health services and the impact this has on the scope of rural nursing practice. The paper will conclude that the size of the health service is an outcome of rurality (small population densities, distance from larger health facilities, lack of on-site medical and allied health staff). It also notes that the size of the health service is a major contextual determinant of patient acuity and staff skill-mix in small rural hospitals, and therefore the scope of rural nursing practice. PMID- 10808785 TI - Evidence-based educational commissioning and policy making: a case for the prospective use of a psychometric approach in defining the role and preparation of the nurse practitioner in Australia. AB - The concept of evidence-based care and the profile of the nurse practitioner (NP) have together assumed mounting importance as possible solutions to the problem of meeting escalating demands for high quality health care with decreasing resources. With regard to the former, concerted efforts to ensure that clinical practice has its base in scientific evidence, have resulted in health care delivery that is increasingly informed by research rather than ritual and tradition. This success notwithstanding, health management and policy-making have remained grounded in assumption, principally because of the paucity of available research in these areas. With regard to the NP, there is little agreement regarding the boundaries of professional practice and preparatory education, both of which need to be defined to some degree at the local level, if the health needs of the indigenous population are to be met. If these two issues are integrated, the policies regarding the preparation and occupational scope of the NP could incorporate research-based decision-making, and thus could simultaneously develop the NP according to locally specified needs, as well as meeting the requirements of the evidence-based care culture. This paper discusses a psychometrically valid and reliable instrument that has the capacity to provide research data which can inform strategic thinking regarding the definition, education and operation of the NP at a variety of levels. The technique has been used extensively in the UK, repeatedly demonstrating its value in NP educational commissioning. While the instrument has not yet been used to inform NP education in Australia, it has been administered to a cohort of acute sector nurses in Victoria. It is argued that the relevance of the instrument to the Australian health system, coupled with its proven importance in the development of the NP in comparable health care cultures, makes this approach worthy of consideration for the preparation of the NP for a variety of Australian health contexts. PMID- 10808786 TI - Nursing's contribution to general practice: general practitioners' and practice nurses' views. AB - This study was instigated by the paucity of knowledge relating to the work of nurses employed by general practitioners in Australia and the phenomenal development of the practice nurse role in the United Kingdom. A mailed survey of general practitioners and practice nurses within one Division of General Practice in South-East Queensland, sought, among other things, their views about the current and potential contribution of nursing to general practice. Eighty-four out of 164 (51%) general practitioners and 37 out of 67 (55%) practice nurses responded to the survey. Results indicated that both general practitioners and practice nurses appreciated the value of nursing services in general practice and would sanction the employment of more nurses especially for the purpose of preventive care. The majority of nurses were agreeable to the notion that the nurse's role could, and should, be expanded to include autonomous functioning while most of the doctors were amenable to some extension of nursing practice but reticent or opposed to any independent interventions. PMID- 10808787 TI - World Wide Web: health informatics. PMID- 10808788 TI - Milestones in Australian nursing. PMID- 10808789 TI - Organisation of patient care in the Roman military hospital. PMID- 10808790 TI - Stewardship. PMID- 10808791 TI - Stewardship includes 'health care without harm'. PMID- 10808792 TI - Stewardship as a quality of nursing practice. Interview by Marie Manthey. PMID- 10808793 TI - Who owns a staff nurse's time? PMID- 10808794 TI - An ecospiritual consciousness. PMID- 10808795 TI - Carrying hope. PMID- 10808796 TI - Calling patients after emergency visits. PMID- 10808797 TI - Creating healing environments. PMID- 10808798 TI - The art of nursing. PMID- 10808799 TI - Nursing's 21st century values. PMID- 10808800 TI - A Nightingale legacy: the art of nursing. Interview by Marie Manthey. PMID- 10808801 TI - The art of nursing: a clinicians' view. Interview by Lisa Legge. PMID- 10808802 TI - Nursing, complementary therapies and the emerging healthcare system. PMID- 10808803 TI - A therapeutic conversation for the caregiver. PMID- 10808804 TI - Holistic nursing--a return to healing. PMID- 10808805 TI - Treatment protocols and pathways: improving the process of care. PMID- 10808806 TI - Have we reached the therapeutic ceiling in acute myocardial infarction? AB - Despite the recent introduction of several new agents into the fibrinolytic market, none have proven more efficacious than earlier agents. However, new routes of administration may provide some benefit in terms of time to treatment and ease of administration. The best possibility for breaking the therapeutic ceiling is in finding new ways to use fibrinolytic agents, such as combination therapy with GP IIb/IIIa inhibitors and low-molecular-weight heparin. Use with PCI may decrease mortality rates in some patient groups. Prehospital administration of fibrinolytics is also an option for expediting care and improving outcomes. PMID- 10808807 TI - The role of the critical care nurse in preventing heart failure after acute myocardial infarction. AB - Heart failure is a disabling, progressive, and highly lethal condition. This profile makes it an ideal target for preventative strategies. Critical care nurses caring for cardiac patients must be aware of these strategies so that they can effectively assess patient risk and intervene as an educator and advocate for appropriate treatment. PMID- 10808808 TI - Preventing prehospital delays in seeking care for acute myocardial infarction: a patient education program. AB - Critical care nurses often lament about patient situations in which their ability to provide definitive nursing care is limited. In the care of the patient vulnerable to AMI, however, a number of concrete interventions are available to ameliorate the potentially devastating effects of this disorder. Implementation of the patient education program described here is one sure way that critical care nurses can assist in effectively reducing the mortality and morbidity associated with AMI. Adopt it and make a difference. PMID- 10808809 TI - Early extubation and early activity after open heart surgery. PMID- 10808810 TI - Percutaneous myocardial revascularization: new treatment option for patients with angina. PMID- 10808811 TI - Naloxone infusion and drainage of cerebrospinal fluid as adjuncts to postoperative care after repair of thoracoabdominal aneurysms. AB - The mechanisms that produce paraplegia in patients after TAA repair are complex and involve alterations in regional blood flow to the spinal cord, CSF dynamics, and reperfusion. Although neither the minimal level of blood flow nor the maximal spinal cord pressure that can be tolerated by the spinal cord is known, adjuncts such as CSF drainage and naloxone infusions may allow longer durations of aortic cross-clamping before irreversible ischemia occurs. Because paraplegia is multifactorial and none of the recommended adjuncts alone provides complete protection of the spinal cord, a combination of treatments may be necessary to reduce the prevalence of neurological complications after thoracoabdominal aortic reconstruction. Critical care nurses thus must be acquainted with the advanced monitoring techniques and the pathophysiology behind these new treatment modalities. Advanced assessment skills are also essential to recognize the potential neurological complications that may occur in these patients. Care of patients with TAA is a challenge. Critical care nurses must use multidimensional skills in the areas of hemodynamic monitoring, physical assessment, and psychological counseling to effectively manage postoperative care of these patients. PMID- 10808812 TI - Frequency of ischemia during intracoronary ultrasound in women with and without coronary artery disease. AB - Myocardial ischemia is common during ICUS imaging in women with and without CAD. Although no long-term adverse effects occurred in our small sample, a larger sample of women is required to confirm our observations and to determine the precise mechanisms of ischemia. Such studies may determine whether the smaller diameter of coronary vessels in women makes the women more vulnerable than men to the occurrence of chest pain and ischemia during ICUS. Although ICUS is valuable in guiding coronary interventions, disposable catheters are costly. Studies are required to assess the cost-benefit ratio of incorporating ICUS with coronary interventional procedures. Until more is known, we recommend that nurses educate patients about ICUS, monitor them closely for ischemia and arrhythmias during the procedure, and consider obtaining 12-lead ECGs when patients undergo and ICUS procedure. PMID- 10808813 TI - Preoperative versus postoperative weights: which one should be used for cardiac surgery patients' drug and hemodynamic calculations? PMID- 10808814 TI - The expanding role of signal-averaged electrocardiography. AB - Signal-averaged electrocardiography is a valuable diagnostic tool for determining which patients recovering from myocardial infarction are at risk of sudden death due to ventricular arrhythmias. Additionally, the value of this technique in determining which patients with ischemic heart disease and unexplained syncope are likely to have inducible sustained ventricular tachycardia has been established. This noninvasive screening procedure has shown promise in other clinical situations, but more investigation is needed before definitive recommendation can be made. Critical care nurses can help promote the success of signal-averaged electrocardiography by educating patients, promoting acquisition of a quality recording, helping allay patients' concerns, and participating in research activities. PMID- 10808815 TI - Blunt chest trauma: case report. AB - Care of patients with blunt cardiac trauma is challenging for bedside nurses because of the potentially elusive and subtle nature of clinical findings associated with such trauma. An understanding of the forces exerted during the trauma episode will assist nurses in the assessment and search for hidden injuries. A high index of suspicion and continued cautious assessment and attention to subtle changes in the patient's physical condition are essential. Keen attention to changes in the patient's vital signs, cardiac output, ECG findings, pulses, and fluid volume status alert nurses to potential injuries associated with blunt cardiac trauma ranging from myocardial contusion to cardiac tamponade, aortic tears, and cardiac rupture. Survival of patients with blunt cardiac trauma depends on early intervention. PMID- 10808816 TI - Advanced practice nurses guiding families through systems. PMID- 10808817 TI - Weaning from short-term mechanical ventilation. PMID- 10808818 TI - Is there a difference in the reliability of results when evaluating the square wave of the pulmonary artery waveform as opposed to that of the arterial waveform? PMID- 10808819 TI - Pediatric and neonatal nurses get "one more hand". PMID- 10808820 TI - Humour--a critical faculty in critical care nursing? PMID- 10808821 TI - Strategies in postoperative pain assessment: validation study. AB - Pain assessment and management are major clinical problems that many categories of healthcare professionals have to deal with. Although there are many potentially successful approaches available for pain management, there is still a shortage of knowledge about the strategies used by staff members for the actual assessment of pain and how reliable these strategies are. The fact that patients often undergo a great deal of suffering from pain and lack of adequate pain relief may be considered an indicator of this shortage of knowledge. Clinical studies from different parts of the world reveal that the incidence of pain reported by patients is still high, with about 75% reporting moderate pain and an additional 15% severe pain. The aim of the present study was to validate different categories used in acute pain assessment and their accuracy in a new clinical sample and to explore further different dimensions of how staff members experience pain assessment. Intensive care nurses (n = 10) were carrying out pain assessment of postoperative patients (n = 30). Each pain assessment was followed by a detailed interview and indicating the estimated pain intensity on a visual analogue scale (VAS, 0-10 cm). The pain ratings by the nurses were compared to those of the patients to assess the accuracy of the pain assessments of the staff members. A previously developed category system for describing the initial empirical material regarding criteria the nurses relied on when assessing pain, combined with what experience has taught them in this respect, was used to assess the validity of previous observations. The results indicate that similar approaches were still used by the nurses but the accuracy of pain assessment had considerably improved. PMID- 10808822 TI - Management of diabetes and hyperglycaemia during myocardial infarction: review of the literature. AB - For many years now, research has firmly demonstrated the increased mortality in patients with diabetes following myocardial infarction (MI), a prognosis which has persisted despite major advances in acute coronary care. Research has also shown higher than usual mortality rates in patients without known diabetes presenting with hyperglycaemia during MI. Due to a lack of research evidence, little has been established about how best to manage glycaemic control in these patients during the acute phase of an MI. However, a recent clinical trial has had considerable impact on coronary care practice. It advocates intravenous insulin therapy for all diabetics and patients with hyperglycaemia during acute MI, followed by subcutaneous insulin for three months, regardless of previous treatment. The evidence for mortality benefit is substantial, but the trial has left some questions unanswered. The aim in this literature review is to examine critically the research basis for using insulin during and after MI, and to identify the potential impact of the research on patients and nurses. The author searched the CINAHL and MEDLINE indexes for relevant texts in English from 1975 to 1998, and has recently implemented relevant knowledge from this research into her own work area, a coronary care unit in the north of England. PMID- 10808823 TI - A Heideggerian investigation into the lived experience of humour by nurses in an intensive care unit. AB - A literature search revealed no systematic enquiry into the experience of humour by nurses in the intensive care setting, suggesting that there was a need for such studies (Astedt-Kurki & Liukkonen 1994). This phenomenological study was undertaken using an interpretive human science approach of phenomenology grounded in the hermeneutics of Heidegger. The task was to investigate and describe the concept of humour in an intensive care unit (ICU) from an inductive-descriptive perspective, seeking to identify the essence of the phenomenon through an accurate description of the lived experience of humour. A sample of eight nurses with at least one year's intensive care experience was randomly selected. Semi structured interviews were conducted, audiotaped, transcribed into text, checked for transcription errors, and returned to participants for checking to ensure reliability and validity. Main core themes were identified and categorized, then Colaizzi's seven steps were used for analysis. Themes and categories identified in the collected data were rich and varied, allowing for lengthy discussions on thoughts and experiences. The study revealed a rich experience of humour in the ICU, suggesting that humour is central to nursing, and therefore worthy of deeper analysis using the same methodology. PMID- 10808824 TI - Paediatric intensive care in a district general hospital. AB - The report of the 'National Coordinating Group on Paediatric Intensive Care' (NHS Executive 1997) recommended that general (adult) intensive care units in district general hospitals should no longer continue to care for critically ill children apart from providing resuscitation prior to transfer to a specialist centre. This recommendation was made despite a lack of outcome data in the UK to support this conclusion. We wish to report our outcome data from one such unit. Over a four year period, we have prospectively used Paediatric Risk of Mortality scoring to estimate the risk of death for 95 children cared for in our adult unit in a district general hospital (DGH) and have compared this with the observed outcome. The observed mortality of 2 deaths was less than the predicted mortality of 2.32 deaths. Our results suggest that an adult unit can provide a paediatric intensive care service that is associated with an acceptable outcome in terms of mortality. These data suggest that the role of such units with regard to the provision of paediatric intensive care should be considered further. PMID- 10808825 TI - Collecting data in the operating department: issues in observational methodology. AB - The UK literature contains few references to nursing in the operating department and even fewer to methods for researching nursing in what is often a complex and stressful environment. Nursing staff work under intense pressure and this presents a challenge, not only for nurses who work in the operating department, but also for researchers who choose to study nursing within its bounds. This paper focuses on some of the key issues which emerged during one study of nursing in the operating department where an observational methodology was used. The author suggests that observation, as a research method, is an important feature of contextual investigation. However, like other methods of investigation, it requires careful planning and preparation. Furthermore, the author believes that sensitivity and skill are required for the enactment of the observer role when researching nursing within this specialized environment. PMID- 10808826 TI - Orlistat (Xenical). AB - The introduction of orlistat (Roche, UK) at the end of last year received considerable media coverage and was heralded in some quarters as the 'quick fix' for those wishing to lose weight without the pain of dieting. This description is far from accurate but treatment with orlistat may have a role in patients for whom obesity is a serious and even life-threatening condition. PMID- 10808827 TI - Critical care nursing in Canada. PMID- 10808828 TI - Chest X-ray quiz. Mitral stenosis. PMID- 10808829 TI - Influence of southern spiritual beliefs on perceptions of mental illness. AB - Cultural religious beliefs influence perceptions of mental illness, and any clinician interested in treating mentally ill people and their families must consider these beliefs so that he or she can develop culturally specific interventions. This article reports on the results of interviews with African American experts, mentally ill persons, and nurses caring for the mentally ill. A case study is used to illustrate the influence of southern religious beliefs on perceptions of mental illness and the behaviors of people who are mentally ill. Although many issues are considered in this analysis (i.e., ethnicity, geographic location, and religion), it is the influence of three religious traditions in the South--voodoo, slave religion, and evangelical Protestantism--that takes precedence in the analysis. Mental health professionals, especially psychiatric nurses, will find this information helpful when assisting hospitalized patients. PMID- 10808830 TI - Appropriately timed interventions with the family and the adolescent: the role of the therapist. AB - Family therapy conducted with a troubled adolescent is a difficult process. In this article the case scenario of a troubled teenager is presented. The adolescent's behavior is scrutinized through an examination of her growth, dynamics, family/parent interactions, and social support. Therapeutic and timely interventions--a blueprint for family therapy integrating psychoanalytic, Bowen, Milan, and behavioral approaches--are explained. The blueprint can be used to assess and assist in the change process of the adolescent. It is extremely important that new therapists realize when they are getting drawn into the content without appreciating the process of therapy. This realization will foster second-order change in the growth of a family therapist. PMID- 10808831 TI - "Living in hell": the experience of being stalked. AB - Stalking is a form of interpersonal victimization that can have profound effects on targeted individuals by necessitating unwanted life changes and instilling fear that may persist long after the stalking has stopped. This article reviews current literature on stalking, presents a case study of a woman who was stalked for 3 years by a near-stranger, and offers recommendations for nurses and other mental health clinicians who work with victims of stalking. PMID- 10808832 TI - Relationship between coping strategies and depression among employed Korean immigrant wives. AB - Coping strategies, such as working harder and negotiation, may have an effect on depression for employed Korean immigrant wives. Additionally, income and education have been associated with depression in previous research. A cross sectional survey research design was used to explore which coping strategies and demographic variables were significantly related to depression for employed Korean immigrant wives. The results of multiple regression analysis revealed that coping strategies and demographic variables accounted for 24% of the variance in depression. Specifically, as wives worked harder cleaning the house, their depression increased, whereas when they negotiated with their husbands, they were less likely to be depressed. To enhance negotiation among Korean wives, mental health nurses need to work within the community to foster the development of cultural and traditional norms that sanction negotiation between husbands and wives. PMID- 10808833 TI - Hospital-based psychiatric experience before community-based practice for nurses: imperative or dispensable? AB - This article describes an Australian research project that explored the relevance of hospital-based experience in preparing psychiatric nurses for community-based practice. A qualitative design was selected to obtain in-depth information in an area in which no formal research has been undertaken. In-depth interviews were conducted with 6 psychiatric nurses currently engaged in community-based practice. The interviews were audiotaped, and the transcribed data were analyzed for major themes. The results indicated that the participants did not believe their hospital experience had prepared them to function effectively in the community. In some respects hospital experience was perceived as having hindered their transition into the community environment. This exploratory study indicates the need for further research and the exploration of alternative methods to prepare psychiatric nurses for community-based practice. PMID- 10808834 TI - Leading through competence. PMID- 10808835 TI - Oncology nurses' use of the Internet for continuing education: a survey of Oncology Nursing Society Congress attendees. AB - A publication and continuing education company conducted a survey at the 23rd Annual Oncology Nursing Society Congress to determine whether nurses are using the Internet and, if so, for what purposes. Survey results revealed that oncology nurses are using the Internet for a variety of educational purposes, which include drug information, literature searches, academic information, continuing education, and patient education. Continuing education providers should continue to pursue the Internet as a means of meeting the needs of oncology nurses for quick, up-to-date information in their field. There is a need for further documentation on the use of the Internet by nurses for continuing education, including topics and program formats that would be most beneficial. PMID- 10808836 TI - The Pyramid Model: an integrated approach for evaluating continuing education programs and outcomes. AB - Recent mergers and downsizing of health care agencies have made resources for continuing education (CE) increasingly scarce. Nurse educators must demonstrate the effectiveness and sustainability of CE programs and establish the link between nursing professionalism and positive patient outcomes. The Pyramid Evaluation Model expands and enhances previous evaluation frameworks. Simple steps are outlined to evaluate CE programs and outcomes systematically and comprehensively through an impact model that examines goals, reviews program design, monitors program implementation, assesses outcomes and impact, and analyzes efficiency. PMID- 10808837 TI - Developing collaborative partnerships in continuing nursing education: walking the talk. AB - The purpose of this article is to describe and analyze a reciprocal partnership developed among four university-based continuing education providers in Wisconsin. Members of this partnership collaborated in developing a statewide program to address the educational needs of nurses moving from acute or long-term care into community nursing. Components of the reciprocal partnership model were applied in developing this program. The partners identified that, despite the initial time investment, the collaborative approach was an advantage. The collaborative approach enhanced the quality of the program developed and reached a wider audience. Continuing nursing educators may want to consider partnership arrangements for program development and implementation. PMID- 10808838 TI - Continuing education needs of nurses employed in nursing facilities. AB - The purpose of this investigation was to determine the self-perceived learning needs of nurses, both RNs and licensed practical nurses (LPNs), employed in nursing facilities. Questionnaires were used to gather data from 319 nurses employed in 14 nursing facilities, representing 10% of all nursing facilities in North Carolina. A total of 164 nurses returned usable questionnaires. The nurses indicated a large number of continuing education needs, with Management Skills, Drug Therapy/Interactions, and Behavioral Problems being particularly important. Night shift nurses and nurse educators had different leaning needs when compared to other subsets of nurses. PMID- 10808839 TI - Substance abuse education liaisons: a collaborative continuing education program for nurses in acute care settings. AB - BACKGROUND: Alcohol, tobacco, and other drug abuse undermine physical and psychological well-being, contributing to the array of illnesses that necessitate admission to acute care settings. Addictive disorders often are undetected, underreported, or overshadowed by the primary illness. Nurses need continuing education to enhance competence in meeting this challenging problem. METHOD: Through a unique collaboration between university faculty and acute care nurses from seven hospitals, the nurses attended 18 monthly workshops designed to meet their learning needs related to substance abuse. RESULTS: The nurses acquired essential knowledge and skills regarding substance abusing clients. The publication of a substance abuse resource manual and the establishment of a continuing network of professionals committed to improving practice are additional positive outcomes of this endeavor. CONCLUSION: Project SAEL (Substance Abuse Education Liaisons) is a model that can be emulated by others. PMID- 10808840 TI - Access all areas: wound care resources on the Internet. AB - The World Wide Web provides access to a plethora of information on every conceivable topic. More and more, the Internet is been used to access health information, making service users better informed about their conditions and the choice of treatment. It is essential that healthcare professionals embrace this technology so that they are able to access information from around the world. Ideally, access to such technology should be as close as possible to the point of care delivery. The expansion of the NHSnet over the next 3-5 years should make this a reality for more and more practitioners. PMID- 10808841 TI - Using a case-mix-adjusted pressure sore incidence study in a surgical directorate to improve patient outcomes in pressure ulcer prevention. AB - The Glasgow Acute Clinical Audit Sub-Committee on Pressure Sores has previously carried out studies of incidence of pressure ulcers in the medical directorates and case-mix-adjusted the figures for length of hospital stay and risk assessment score. Case-mix classification is 'classification of people or treatment placed into groups using characteristics associated with condition, treatment or outcome that can be used to predict need, resource, use of outcomes'. In this instance, crude pressure ulcer incidence figures may be adjusted for length of hospital stay and pressure sore risk assessment score, and stratified into groups, which allows like to be compared with like. The value in case-mix-adjusted figures lies in repeating the exercise, thus determining the trend for individual areas and assessing whether improvement in the quality of care is being achieved. This is more positive than creation of 'league tables' comparing simultaneous studies in a number of areas. The figures showed that there was no statistically significant difference between surgical directorates in trusts with regard to risk assessment scores and length of hospital stay. Gathering data on the incidence of pressure ulcer development allows us to identify where new sores are occurring, but does not critically analyse the nursing intervention taken in individual cases, which identifies preventive strategies. The Glasgow group's primary aim was to gather data on case-mix-adjusted incidence of pressure damage; the secondary objectives were to scrutinize the data to gather more general information on intrinsic and extrinsic factors which may predispose to pressure ulcer development. The study was carried out in the surgical directorate. Findings showed that incidence was low (1.1%), with the majority of sores being superficial. There was a correlation between pressure ulcer development and incontinence, evidence of under utilization of moving and handling aids for prevention of pressure ulcers, and a need for greater collaboration between nursing staff and dietitians. All patients had access to pressure-reducing surfaces, with cut-foam pressure-reducing mattresses with vapour-permeable covers on all beds. Dynamic systems were available if the primary nurse identified a need for them. PMID- 10808842 TI - The anti-microbial activity of maggot secretions: results of a preliminary study. AB - The ability of larval secretions to kill or prevent the growth of a range of potentially pathogenic bacteria was investigated in a preliminary laboratory study. Marked anti-microbial activity was detected against Streptococcus A and B and Staphylococcus aureus. Some activity was also detected against Pseudomonas sp. and a clinical isolate of a resistant strain of S. aureus (MRSA). No evidence of inhibition was recorded against Enterococcus or the Gram-negative bacteria Escherichia coli and Proteus. These findings are consistent with clinical observations made by us and others that larvae of the common greenbottle are able to combat clinical infections in a variety of wound types including those caused by antibiotic-resistant strains of bacteria. PMID- 10808843 TI - The contribution made by an armchair with integral pressure-reducing cushion in the prevention of pressure sore incidence in the elderly, acutely ill patient. AB - The paper describes a clinical controlled trial of an armchair with integral pressure-reducing cushion, which took place on two elderly acute medical wards in a district general hospital. The aim of the study was to evaluate whether the armchair would have an impact on reducing the occurrence of pressure sores. All patients were provided with a mattress or overlay in accordance with the hospital policy on mattress provision following a Waterlow risk assessment. Patients on ward A were provided with an armchair with integral pressure-reducing cushion, whilst patients on ward B continued to use their existing bedside armchair. Nominal data were collected on all pressure sores in patients who were not bedridden. Ward A had a significantly lower incidence of hospital-acquired pressure sores compared to ward B, and non-hospital-acquired pressure sores on this ward showed more improvement than those on ward B. PMID- 10808844 TI - Beds, mattresses and cushions for pressure sore prevention and treatment. PMID- 10808845 TI - Local interventions for pain in venous leg ulcers. PMID- 10808846 TI - The choice of products in wound management. PMID- 10808847 TI - Diabetic foot ulcer associated with Wegener's granulomatosis. PMID- 10808848 TI - Management of chronic venous leg ulcers using a new autologous skin graft system. PMID- 10808849 TI - The assessment and treatment of wound pain. PMID- 10808850 TI - Prescribing for leg ulcers in general practice, Part 2. AB - This study, the second of a two-part article, reports the findings of a survey undertaken to establish who selects the product used in the management of leg ulcers, and the factors influencing their decision making. The use of interviews (n = 9) followed by postal questionnaires (n = 197) confirms that the majority of decisions are made by practice nurses and district nurses. The findings suggest that, of the factors considered, patient comfort and compliance are the most important factors influencing product selection. This study also suggests that nurses are not using the best available evidence because of inaccessibility of resources and lack of time to search the literature. Nurses also cited difficulty in discriminating between a biased presentation and reliable research. PMID- 10808851 TI - Adsorption of serum-derived proteins by primary dressings: implications for dressing adhesion to wounds. AB - Using an in vitro immunolocalization technique, an exploratory study was carried out into the serum-derived protein adsorption capacity and the cell adherence of a traditional gauze dressing versus a new gelling fibre gauze dressing. We found that the traditional gauze dressing adsorbed protein more readily than the new dressing. The findings indicate that reduced binding of serum proteins to the surface of the gelling fibre dressing may help reduce the adherence characteristics for this type of dressing, minimising trauma and possibly reducing the acute pain experienced during dressing changes. PMID- 10808852 TI - The management of chronic wounds: factors that affect nurses' decision-making. AB - This study sought to determine how registered nurses make decisions regarding the management of chronic wounds. A self-administered questionnaire consisting of both Likert-type and open-ended questions was used to survey registered nurses working in hospitals and community services in a large Australian city. A total of 140 questionnaires were returned from a variety of clinical settings. The study revealed that registered nurses had a significant role in chronic wound management. However, they relied primarily on their own experience and that of colleagues for decision making in which they had varying degrees of autonomy. The authors conclude that nurses and medical staff require more objective, research based education on wound assessment and management. PMID- 10808853 TI - The role of honey in the management of wounds. PMID- 10808854 TI - The management of diabetic foot disease in Finland. PMID- 10808855 TI - On fathers. PMID- 10808856 TI - Misoprostol. PMID- 10808857 TI - Misoprostol. PMID- 10808858 TI - Fathers can make a difference. PMID- 10808859 TI - Bravo, fathers! Even when we don't fully understand them.... PMID- 10808860 TI - Birth fathers or knaves-in-waiting? PMID- 10808861 TI - The role of the father in childbirth. PMID- 10808862 TI - Unassisted homebirth: one father's experience. PMID- 10808863 TI - Caught off guard: one father's memories. PMID- 10808864 TI - A father's point of view. PMID- 10808865 TI - Making decisions, providing comfort. PMID- 10808866 TI - A note to fathers: it's you she wants. PMID- 10808867 TI - Is the participation of the father at birth dangerous? PMID- 10808868 TI - Child is father to the man. A survivor of childhood sexual abuse speaks out. PMID- 10808869 TI - Painful memories: the midwife's role in helping fathers. PMID- 10808870 TI - Pregnancy from a new perspective. PMID- 10808871 TI - Ultrasound. Weighing the propaganda against the facts. PMID- 10808872 TI - My journey to natural family planning. PMID- 10808873 TI - Gladys Milton, Florida midwife. May 26, 1924-June 17, 1999. PMID- 10808874 TI - The vocation of Shiphrah. PMID- 10808875 TI - A wait and see approach with marginal placenta praevia. PMID- 10808876 TI - Evidence-informed midwifery. 1. What is evidence-informed midwifery? PMID- 10808877 TI - Breastfeeding discrimination and censorship. PMID- 10808879 TI - Maternal healthcare in Chile. PMID- 10808878 TI - Labor of love to improve birthing in the Caribbean. PMID- 10808881 TI - Home Birth Association of Ireland. PMID- 10808880 TI - A waterbirth that turned the tide. PMID- 10808882 TI - Meet Minna Jestila, Finnish nurse in Norway. PMID- 10808883 TI - Tasmanian angel: Jane Hall. PMID- 10808884 TI - Newborn genetic screening: blessing or curse? AB - Newly discovered genes and advances in genetic screening programs prompt many questions reflecting the kinds of ethical dilemmas that go hand in hand with life changing discoveries. Neonatal genetic screening has been a standard of care for some time, but as our knowledge in the field of genetics expands, should we continue with the same approach? What newborn genetic screening tests should be mandatory, and what are the long-range consequences associated with testing? This article reviews genetic modes of inheritance, outlines and explains the most common newborn screening tests, and enumerates the ethical issues associated with these screening procedures. The role of the neonatal nurse in the newborn genetic screening process is discussed. PMID- 10808885 TI - Cisapride: the problem of the heart. AB - Cisapride has been used in the neonatal population as a first-line gastrointestinal prokinetic agent for managing feeding intolerance secondary to decreased gastrointestinal motility. Cisapride acts specifically at many levels of the gastrointestinal tract and has no central nervous system side effects. Recently, prolonged QT interval has been reported secondary to high-dose cisapride, and concerns regarding its use in premature infants have been raised. In response, Janssen-Ortho Inc. and Janssen Pharmaceutica, the manufacturers of cisapride in Canada and the U.S., have released safety information prohibiting the use of cisapride in premature infants. In light of the present evidence, it is imperative that NICUs that continue to prescribe cisapride for the management of feeding intolerance exercise vigilance in cardiac monitoring and recognize the importance of drug interactions that lead to elevated cisapride levels. PMID- 10808886 TI - Development of father-infant attachment in fathers of preterm infants. AB - PURPOSE: To investigate the development of feelings of attachment between fathers and their preterm infants and to identify factors that help or hinder this process. DESIGN: A longitudinal descriptive design was used to obtain fathers' perceptions of their infants, feelings for their infants, and other related factors. SAMPLE: A convenience sample of 27 fathers of preterm infants was recruited. MAIN OUTCOME VARIABLE: The main outcome variable was the time at which fathers first held their infants. RESULTS: The earlier fathers held their babies, the sooner they reported feelings of warmth and love for them. PMID- 10808887 TI - Caring for the visually impaired infant. PMID- 10808888 TI - Use of surfactant for respiratory distress syndrome. PMID- 10808889 TI - Workforce changes: trends and implications for neonatal nurses. PMID- 10808890 TI - Problem-based learning in the NICU. AB - Problem-based learning has been used by individuals from various professions and in a wide array of academic and clinical settings. The primary benefits of PBL are that it allows learners to be actively engaged in the learning process and that it can help to strengthen problem-solving skills for learners on any level. PBL can be used effectively in the NICU for both novice and advanced practice nurses. PMID- 10808891 TI - Playing the team-building game. PMID- 10808892 TI - Primacy of the printed word: is it under threat? PMID- 10808893 TI - Computer assisted learning: the potential for teaching and assessing in nursing. AB - This article discusses computer assisted learning (CAL) and the importance of applying it in nurse education. The articles recognizes the general technological developments as exemplified by the Teaching and Learning Technology Programme (TLTP) from which ideas about application and benefits came. The ideas from TLTP are hereby used in CAL and applied to nursing and health-care undergraduate programmes in one university. In the light of this experience the main intention of this article is to consider the benefits and costs of introducing computer programmes as part of the teaching provision for nurses and other health-care professionals both at beginner and advanced level. The article further argues that CAL can also be used for patient teaching thus providing transferable skills and benefits for teachers as well as learners, be they students or patients. To support such multiple uses of CAL selected examples will be offered and appropriate conclusions will be drawn. PMID- 10808894 TI - Integrating the creative arts into a midwifery curriculum: a teaching innovation report. AB - The practice of midwifery has long been recognized as both art and science. However, educational programmes for midwifery are most often undertaken within an academic health sciences environment, and tend to be based on knowledge derived from the sciences (e.g. life sciences, biomedical sciences, behavioural sciences and social sciences). These scientific perspectives, while essential to the preparatory and on-going education of midwives, do not necessarily fully prepare midwives to fulfil their practice roles. This paper reports a teaching innovation aimed at facilitating student exploration of fundamental, complex and ethereal concepts which are essential to the effective and skillful practice of midwifery. Through the exploration of the arts and humanities, students were encouraged to engage with concepts such as 'caring', 'empathy', 'suffering', 'motherhood', 'pain', 'love', 'attachment', 'health' and 'illness'. Students were also encouraged to explore cultural and social symbols pertaining to parenthood and family life. Evaluation revealed that students valued the course, and that they gained insights which assisted them to develop understanding of key concepts. Implications for practice and education are drawn from this paper. PMID- 10808895 TI - Student nurses' conceptions of internationalization: a phenomenographic study. AB - A phenomenographic approach was used when analysing and interpreting interviews with 25 student nurses in order to describe their conceptions of internationalization. The results are presented in five categories and various subcategories, forming an outcome space. The categories describe forms of understanding, or ways of thinking about internationalization, which, in relation to internationalization as a whole, were either atomistic or holistic in character. The data revealed that nursing students shared some aspects of meaning when thinking of internationalization. These aspects seemed to be understood in a similar way by all students, whereas, when students thought about internationalization as a whole, they assigned it different meanings. In two of the categories aspects of meaning were fragmentarily related to each other, and represented only one perspective. These two categories are to be compared to the other three categories which were more complex and holistic in character, since they contained more than one perspective as well as more aspects of meaning, and related in several different ways. From a pedagogical point of view, the findings point to the importance of teachers trying, in an active way, to confront students with as many existing variations of conceptualized wholes related to internationalization as possible, by using forms of understanding originating from a holistic approach. PMID- 10808896 TI - Considering academic qualification in mathematics as an entry requirement for a diploma in nursing programme. AB - A study carried out with first year students on a Diploma in Nursing programme at Nottingham University School of Nursing revealed that some students were struggling with basic arithmetic. A group was subsequently set up to address a range of issues relating to mathematics in nursing and nurse education. One of the areas investigated by the group was minimum entry requirement to the Diploma programme. In particular we wanted to determine the reliability of Mathematics GCSE Grade C as an indicator of arithmetic competence. Academic grades in mathematics (achieved prior to attendance at the School of Nursing) were compared with scores in an arithmetic test administered to students upon commencement on the Diploma in Nursing programme. It was found that the performance of students with Mathematics GCSE Grade C varied widely and some students could not solve basic arithmetic problems without the aid of a calculator. Schools of Nursing which adopt a policy of minimum qualifications in mathematics should not be complacent in assuming that students will be competent at arithmetic. The implementation of a programme of early assessment and tutorial support throughout the Diploma in Nursing programme is also recommended. PMID- 10808897 TI - Beyond the rhetoric of problem-based learning: emancipatory limits and links with andragogy. AB - Problem-based learning (PBL), sometimes referred to as enquiry-based learning, is an approach to education that has gained increasing usage within health care in recent years. Its origins very much lie within medical education. The bulk of literature on PBL is optimistic about its potential, especially in relation to nurse education. It is argued here that the benefits of PBL are that it moves toward student-centred education and process-oriented methods that have been taking place for at least 16 years. There are clear links with andragogy although this is not always acknowledged, but the potential move away from emancipatory education inherent in PBL if used without reflection, is inconsistent with andragogy. This article takes a more critical view of the concept and argues that there are significant limits which need to be considered carefully. Apart from the possible move away from the emancipatory aims of education, there is commonly an implicit support of the medical model within PBL which is inappropriate at a time when the limits of medicine are becoming increasingly clear. It is concluded that further debate and research on the approach is necessary, but that as a facilitative strategy PBL does hold some promise. However, it would be inappropriate to use it as a curriculum model if only because it lacks the diversity required of a postmodern curriculum and would not respond effectively to differing student learning styles. PMID- 10808898 TI - Novice and expert perceptions of psychological care and the development of psychological caregiving abilities. AB - Despite the wealth of research pointing to the centrality of psychological care in illness, there appears to be a lack of consensus concerning its nature and the means by which psychological caregiving abilities are acquired. This has an impact on nurse educators, who must identify and harness the perceptions which students hold, and identify the processes which might contribute to their skill development, as they progress from 'novice' to 'expert'. The aims of this small qualitative study were to compare novice and expert perceptions of the nature of psychological care, and to identify those processes that may contribute to nurses' ability to apply psychological understanding in practice. Data collection methods comprised interactive interviews with qualified and novice student nurses, which were analysed thematically using an adaptation of Colaizzi's (1978) reductive procedure. Results suggest that, whilst there is considerable overlap in themes identified between the two groups, novices consider a broader range of themes to be central to psychological caregiving than do experts; similarly, novices identify more contributory factors than do experts in explaining the process of acquiring skill in psychological caregiving. Possible reasons for these differences are considered. Whilst this study is limited in size and scope, the ultimate goal is to utilize central themes within a model of psychological caregiving, which might inform nurse education. In this way, educational programmes might be appropriately focused upon students' needs within the context of real clinical experience. PMID- 10808899 TI - Role of the research ethics committee. AB - Research ethics committees have an important role to play in ensuring the ethical standards and scientific merit of research involving human subjects. There are three important obligations placed on the ethics committee. Firstly, and most importantly, the ethics committee must ensure that the rights of research participants are protected. This is achieved by ensuring that individuals receive sufficient information, which can be easily understood, and ensuring that appropriate strategies are in place to protect participants from potential adverse consequences of the research. Secondly, the research ethics committee has an obligation to society which provides the resources for research and will ultimately be affected by the results. Thirdly, the research ethics committee has an obligation to the researcher. The research proposal should be treated with respect and consideration. The research ethics committee should strive to meet each of these obligations. All researchers should welcome the contribution made by research ethics committees to the research process because they help to ensure that research meets the high ethical and scientific standards expected by society. PMID- 10808900 TI - Back to basics: exploring the conceptual basis of nursing. AB - It has long been recognized that nursing is essentially a practice-based discipline, underpinned by fundamental concepts which inform practice. The purpose of this paper is two-fold: first, to discuss the need for an exploration of these concepts of nursing as a foundation to pre-registration nursing programmes. Second, by utilizing a continuous quality improvement perspective, an evaluation of teachers' experiences with a cohort of adult and child health branch students is considered. It is concluded that a significant proportion of students found that a raised conceptual awareness served to promote theory practice links. PMID- 10808901 TI - Student discontinuations: is the system failing? AB - Are nurse educators guilty of hypocrisy? While espousing holism and evidence based practice, we may be neglecting to support the most distressed and vulnerable in our care--students. Student discontinuation arouses strong emotions, which need to be expressed and resolved. These are often unresolved leaving students and educators frustrated and alienated in 'failure'. Although it cannot be expected that all students will succeed as professionals and academic standards must be maintained, those who fail have a right to expect fairness. With the advent of Subject Quality Review (formally TQA) institutions of higher education may be asked to demonstrate their operational mechanisms for 'managing' the process of student failure. Now would be the best time for departments of nurse education to reflect on experience, review attrition rates and learn from each other in order to establish a model of best practice. Such a review may generate the formulation of national guidelines, safe guarding against the looming threat of litigation. PMID- 10808902 TI - Creating tension: undergraduate student nurses' responses to a problem-based learning curriculum. AB - Qualitative data were collected from undergraduate student nurses (n = 45) who were participating in a problem-based learning programme of education. Data analysis, using a modified grounded theory method, revealed that although it was perceived that there were considerable benefits to be gained from problem-based learning, there were also some disadvantages. A category that was labelled 'creating tension', which consisted of two sub-categories, namely 'making the transition' and 'remembering the aims' emerged from the data. Making the transition highlighted the difficulty in moving to problem-based learning from more traditional methods of education, whilst remembering the aims described an emphasis on the importance that students place on knowledge acquisition. A number of recommendations for educational practice and research are made as a result of these findings. PMID- 10808903 TI - Nursing process: paradigm, paradox or Pandora's box? PMID- 10808904 TI - A response to Rolfe's reply to Closs and Draper. PMID- 10808905 TI - Claiming our international place. PMID- 10808906 TI - Prescribing rights improve practice. PMID- 10808907 TI - Advancing neonatal nursing practice. PMID- 10808908 TI - Debating advanced nursing practice. PMID- 10808909 TI - Examining trends in workforce planning. PMID- 10808910 TI - Community-acquired pneumonia. PMID- 10808911 TI - Are there any new strategies to increase the success rate for smoking cessation? PMID- 10808912 TI - Self-efficacy in managing dyspnea in COPD. PMID- 10808913 TI - [Therapy of pain--a nursing duty?]. PMID- 10808914 TI - [Pain--how it affects quality of life]. PMID- 10808915 TI - [Pain management in children]. PMID- 10808916 TI - [Ethical responsibility in pain management]. PMID- 10808917 TI - [Basic stimulation in geriatric care]. PMID- 10808918 TI - [The nurse on the way to customer orientation--the patient to be considered as customer]. PMID- 10808920 TI - [When nursing care becomes restraint]. PMID- 10808919 TI - [Relaxing the patient by using autogenous training and progressive relaxation]. PMID- 10808921 TI - More guts ... on the floor! PMID- 10808922 TI - Low-cost ways to access the Net. PMID- 10808923 TI - When the Dx is myasthenia gravis. PMID- 10808924 TI - Nursing takes center stage. PMID- 10808925 TI - Killed in school! PMID- 10808926 TI - Breast cancer. PMID- 10808927 TI - Breast reconstruction. PMID- 10808928 TI - The disaster that took our ED by storm. PMID- 10808929 TI - DVT. What every nurse should know. PMID- 10808930 TI - Support surfaces and specialty beds. Part 2: Aggressive pressure relief. PMID- 10808931 TI - Can "healthy" bacteria ward off disease? PMID- 10808932 TI - Chemical hazards. How to protect yourself. PMID- 10808933 TI - In-office testing: a survey. PMID- 10808934 TI - Using eye drops. PMID- 10808936 TI - [The project for the life in the institution: setting standards]. PMID- 10808935 TI - [Quality of life and projects for life at an institution]. PMID- 10808937 TI - [Example of the "Chateau du Loup" (Wolf's Den") residence]. PMID- 10808938 TI - [Emergencies: care of thoracic pain in an elderly subject]. PMID- 10808939 TI - [Meat, is it necessary in the diet of the elderly?]. PMID- 10808940 TI - [Sense organs and the quality of life of the aged person]. PMID- 10808941 TI - [Depressive states in the elderly]. PMID- 10808942 TI - [Temporo-spatial disorientation in the aged]. PMID- 10808943 TI - [Medico-social decisions: aging and staying at home]. PMID- 10808944 TI - [The child and cancer--at the beginning of the year 2000]. PMID- 10808945 TI - [Prognosis and diagnosis of a case of acute leukemia]. PMID- 10808946 TI - [Body image of a child with leukemia]. PMID- 10808947 TI - [Constructing together the management of children]. PMID- 10808949 TI - [Being a nurse for children with cancer]. PMID- 10808948 TI - [An information guide adapted to the parents' needs]. PMID- 10808950 TI - [The proper role of the nurse in case of pain proved by treatment]. PMID- 10808951 TI - [Creativity as a response to illness]. PMID- 10808952 TI - [When a child suffers from the benefits of treatment]. PMID- 10808953 TI - ["I would like to eat like the adults"]. PMID- 10808954 TI - Putting first things first. PMID- 10808955 TI - Perianesthesia care of the transplant patient. PMID- 10808956 TI - Competency-based orientation (CBO). PMID- 10808957 TI - Medical tattooing. PMID- 10808958 TI - A glimpse into the new millennium: a new era for health care. PMID- 10808959 TI - Preadmission testing today. PMID- 10808960 TI - Deaf patients in the OR: a mile in someone else's shoes. PMID- 10808961 TI - The changing face of staffing--UAPs. PMID- 10808962 TI - Cutting costs without cutting corners: a road map to OR savings. PMID- 10808963 TI - Rx for overcoming adversity and focusing on the future. PMID- 10808964 TI - Patient positioning: snow fun in the OR. PMID- 10808965 TI - Art & science. PMID- 10808966 TI - Kinematics of a total arthroplasty of the ankle: comparison to normal ankle motion. AB - Although the concept of a total ankle arthroplasty has been advanced as a method for treating severe ankle arthritis, the clinical experience with all of the models developed has been discouraging. Both the constrained designs, which maximize joint contact area by restricting the available motion, and the unconstrained designs, which allow more normal motion at the expense of higher contact stresses, uniformly result in implant loosening, pain, and clinical failure in 2 to 7 years. This has led to the recommendation against the use of a total ankle arthroplasty except in very low-demand patients. Failure of ankle implants can be ascribed to either anatomic considerations (e.g.--the talus is too small to accommodate the stress transfers of a prosthesis), or mechanical etiologies. Abnormal 3-dimensional motion of the ankle following arthroplasty would fall into the latter category. This study examined the motion that occurs after implantation of an unconstrained-type total ankle arthroplasty. Using previously validated methodology, axially loaded ankle specimens were cycled through an arc of plantarflex/dorsiflexion while measuring the resulting coupled internal/external and varus/valgus rotations. The average coupled motions in prosthetic ankles were not significantly different than their intact controls. There was, however, a significantly increased amount of hysteresis (defined as the difference between the upper and lower pathways of coupled motion at any given sagittal position) that occurred as the ankle was dorsiflexed and plantar flexed. The increased hysteresis was seen in both the axial and coronal planes. This indicates that there was a greater permitted envelop of motion in the prosthetic ankles compared to normal ankles. It is hypothesized that this subtle change in ankle kinematics caused by the arthroplasty leads to abnormal stress transfer at the prosthesis-bone interface, thereby promoting early implant failure. PMID- 10808967 TI - Increased bone mineral density adjacent to hydroxy-apatite-coated ankle arthroplasty. AB - We measured bone mineral density (BMD) in patients treated with a unilateral hydroxy-apatite-coated ankle arthroplasty. The study comprised 11 consecutive patients measured preoperatively and postoperatively after 3, 6, and 12 months (group I) and 17 patients measured once at 12 to 82 months follow-up (group II). BMD was measured in the distal tibia adjacent to the prosthesis and in the calcaneus. BMD of the calcaneus was measured bilaterally as an indicator of changes in foot load. In group I BMD was significantly increased in the distal tibia after 6 and 12 months compared to preoperative values. In group II BMD of the distal tibia was significantly higher compared to the non-operated side. No radiolucencies were detected during follow-up in any case. The increase in BMD and the radiographic findings after uncemented ankle arthroplasty indicates that it is being loaded by the prosthesis. This may indicate a well fixed prosthesis. PMID- 10808968 TI - Salvage first MTP arthrodesis utilizing ICBG: clinical evaluation and outcome. AB - Twelve patients (12 feet) underwent salvage first metatarsalphalangeal (MTP) arthrodesis with structural, interposition autologous iliac crest bone graft (ICBG). Eight patients had a bony defect secondary to failed first MTP joint implant arthroplasties, two had avascular necrosis (AVN) after failed bunion surgery, one had a nonunion of an attempted arthrodesis for failed bunion surgery, and one had been treated for osteomyelitis after cheilectomy. Eleven of the cases had a single dorsal plate secured by screws and one case had two plates, one dorsal and one medial. A plate, crossed screw(s) and/or K-wire combination were chosen in four cases. Indications included first MTP joint pain, metatarsalgia, intractable plantar keratoses (IPK), as well as a functionally and cosmetically short first ray refractory to non surgical management. Average preoperative shortening was 8.5 mm (range 5-17). Clinical arthrodesis was achieved after an average of 12 weeks (range 4-20). Radiographic arthrodesis was achieved in eleven of twelve feet at an average of 15 weeks (range 8-28), with one pseudoarthrosis. AOFAS forefoot clinical rating score averaged 70 points (max 90 after first MTP arthrodesis) at an average follow-up of 22 months (range 5 70). Sesamoiditis, prominent hardware and scar sensitivity were prevalent complaints in four patients postoperatively. Two cases required flap coverage for skin necrosis. Relief of metatarsalgia, good hallux alignment as well as improved patient satisfaction and function were achieved in all cases. There was no symptomatic progression of interphalangeal degenerative change postoperatively. PMID- 10808969 TI - Result of arthrodesis of the hallux metatarsophalangeal joint using bone graft for restoration of length. AB - We treated 24 patients (18 women, six men; average age, 46.4 years; (range, 28 to 66 years) with fusion of the hallux metatarsophalangeal (MTP) joint using bone graft for the restoration of the length of the first ray. This procedure was performed after bone loss subsequent to previous surgeries for the correction of hallux valgus and hallux rigidus with: silastic arthroplasty (11), bunionectomy and distal metatarsal osteotomy (six), Keller resection arthroplasty (five), and total joint replacement (two). The indication for performing the arthrodesis with bone graft was a short first metatarsal, and associated metatarsalgia of the lesser metatarsals in addition to a painful MTP joint with or without deformity. This bone loss was associated with avascular necrosis of the first metatarsal (nine patients) and with osteomyelitis (seven patients). Of the 24 patients, 14 underwent additional concurrent surgery for correction of hammer toes (10), excision of a Morton's neuroma (two), and lesser metatarsal osteotomy (two). All patients were examined clinically and radiographically at a mean interval of 62.7 months after surgery (range, 26 to 108 months). The patients were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) hallux and MTP 100-point outcome scale. Arthrodesis occurred in 19/24 patients (79.1%) at a mean of 13.3 weeks (range, 11 to 16 weeks), and the first ray was lengthened by a mean of 13 mm (range, 0 to 29 mm). Of the five nonunions, two were asymptomatic, and three were subsequently revised successfully, with arthrodesis occurring at a mean of 10.7 weeks. Complications included one deep infection requiring intravenous antibiotics for treatment of osteomyelitis and two minor superficial wound infections. The mean AOFAS score improved from 39 points (range, 22 to 60 points) to 79 points (range, 64 to 90 points). All patients were satisfied with the final outcome of treatment and stated that they would undergo the surgical procedure again. We concluded that arthrodesis of the hallux MTP joint with bone graft to restore bone loss and length of the first ray may be a worthwhile procedure despite the technical difficulty and the high rate of nonunion. PMID- 10808970 TI - A technique for isolated arthrodesis for inflammatory arthritis of the talonavicular joint. AB - There are few reports in the literature documenting the efficacy of isolated arthrodesis for inflammatory arthritis of the talonavicular joint. Accordingly, we reviewed a single surgeon's experience with this procedure in twenty consecutive cases from this patient population. A technique using indirect joint distraction and the combined use of screw and staple fixation was employed. Solid arthrodesis was noted radiographically in 19 of 20 feet (95%) at an average of 11 weeks. Complications included one non-union, one deep venous thrombosis, and one superficial wound infection. Objective results were graded as excellent in 16 cases, good in 3 cases, and poor in one case. Subjectively, 18 patients were satisfied and one patient dissatisfied with the results of the procedure. It is concluded that isolated arthrodesis is an effective procedure for the treatment of inflammatory arthritis of the talonavicular joint, offering significant pain relief and improved function. Additionally, the use of indirect joint distraction and fixation with screws and staples is a reliable technique associated with an excellent fusion rate. PMID- 10808971 TI - Closed ankle fractures in the diabetic patient. AB - Systemic and local manifestations of diabetes mellitus may complicate the treatment of ankle fractures in the diabetic population. We studied 98 patients (73 non-diabetics and 25 diabetics) who were treated for closed ankle fractures by either surgical or non-surgical methods. We found that overall, the risk of infection in the diabetic population (32%) was 4 times higher than in the non diabetic population (8%). The infection rate in the diabetic group treated surgically more than doubled that in the non-diabetic group. Four out of six diabetic patients treated with cast became infected compared to no infections in the five non-diabetics treated with a cast. Even though the diabetic foot and ankle are well studied, the medical literature is not conclusive regarding the management of ankle fractures in the diabetic patient. Diabetic patients treated conservatively had a tendency to become infected over those treated surgically. Peripherovascular disease, peripheral neuropathy and swelling and/or ecchymosis increased the risk of infection in the diabetic population. Diabetic patients with poor compliance had a tendency to become infected more than those who were compliant. We concluded that the diabetic patient who is poorly compliant with evidence of neuropathic disease, peripherovascular disease and severe swelling and ecchymosis presents the most difficult group to manage. Although these patients are poor surgical candidates, they are also the most difficult to manage and also most prone to infection and complications if treated conservatively. When faced with this difficult scenario a multidisciplinary team approach would probably yield the best possible results by early identification and intervention in these patients. PMID- 10808972 TI - The MABAL shoe, an alternative method in contact casting for the treatment of neuropathic diabetic foot ulcers. AB - INTRODUCTION: Total Contact Casting (TCC) is considered the gold standard in the treatment of neuropathic diabetic foot ulcers. To overcome some disadvantages of TCC we developed a removable fiberglass combicast shoe--the MABAL shoe. PATIENTS AND METHODS: The MABAL shoe was used to treat 23 plantar ulcers. RESULTS: Before treatment the mean surface area was 2.0 cm2 (0.5-7.1 cm2). Twenty-one of 23 ulcers healed, with a mean healing time of 34 days (7-75 days). CONCLUSION: The MABAL shoe provides healing of neuropathic diabetic foot ulcers comparable to existing methods of treatment, while offering some potential advantages. The main advantages are mobilization of the ankle, removability of the cast and a less time consuming form of treatment. PMID- 10808973 TI - Plantarflexion torque following reconstruction of Achilles tendinosis or rupture with flexor hallucis longus augmentation. AB - Nine patients treated surgically for Achilles tendon rupture (7 patients) or tendinosis (2 patients) with primary repair or debridement and augmentation with the flexor hallucis longus muscle-tendon unit were evaluated at a mean of 19 months postoperative. Subjective evaluation revealed a high level of satisfaction. All patients returned to work and only two patients reported limitation in their recreational activities. The mean post-operative AOFAS Ankle Hindfoot Score was 90 points. Four patients reported mild occasional pain and one patient complained of moderate daily pain. Motion assessment showed a 20% increase in the hallux MTP dorsiflexion compared to the non-operative side (p = 0.045). No difference in ankle motion was noted. Cybex II+ dynamic evaluation of plantarflexion peak torque was complete on both extremities. The torque deficit on the reconstructed extremity was 20% (p = 0.01) at 120 degrees per second and 26% (p = 0.003) at 30 degrees per second. There is no significant difference between the torque deficit recorded for patients with Achilles rupture and those with Achilles tendinosis. A trend toward improved torque production with longer follow up was observed. PMID- 10808974 TI - Relationship between calf muscle size and strength after achilles rupture repair. AB - The object was to study the relationships between calf muscle size and strength in 85 patients an average of 3.1 years after repair of achilles tendon rupture. The isokinetic calf muscle strength results were excellent or good for 73% of the patients, whereas calf muscle size was normal in only 30%. The average plantar flexion peak torque per unit muscle cross-sectional area was higher on the injured side than on the uninjured side. The average calf muscle cross-sectional area deficit was 15+/-9% (p<0.001) of that on the unaffected side, while the average plantar flexion peak torque deficit was speed-dependent, being 9+/-18%, 10+/-18 and 2+/-13% of that on the unaffected side at 30, 90, and 240 degrees/sec (p<0.001). The correlation between cross-sectional area and peak torque varied in the range 0.52-0.61 at 30, 90 and 240 degrees/sec (p<0.001). PMID- 10808975 TI - The mobility of the proximal tibio-fibular joint. A Roentgen Stereophotogrammetric Analysis on six cadaver specimens. AB - In six cadaver specimens the mobility of the proximal fibula in relation to the tibia was investigated during plantar/dorsiflexion of the ankle, using Roentgen Stereophotogrammetric Analysis (RSA). The role of the ankle joint, and of the calcaneofibular and talofibular ligaments was also evaluated. The greatest movements were observed along the mediolateral and anterior-posterior axes, resulting in an anterolateral displacement of the fibula head during dorsiflexion and in a postero-medial displacement during plantarflexion. This study demonstrated a limited mobility of the proximal tibiofibular joint. Moreover, the ligament cutting and the presence of constraints in the ankle region did not show any effect on the fibular movement. PMID- 10808976 TI - A determination of ankle kinematics using fluoroscopy. AB - In vivo weight-bearing studies utilizing dynamic video fluoroscopy have been shown to offer an accurate and reproducible method for determining the kinematics of a joint. The purpose of this study was to evaluate translational and rotational motions of the distal tibia relative to the talus in the sagittal and frontal planes. Ten subjects, each having a normal ankle and a total ankle arthroplasty on the opposite side (Buechel-Pappas Total Ankle, Endotec, South Orange, NJ), were studied under in vivo, weight-bearing conditions using video fluoroscopy. All ten subjects were judged to have a successful arthroplasty without demonstrable pain or ligament instability. Under weight-bearing conditions, each subject performed successive motions moving from maximum dorsiflexion to plantarflexion. At maximum dorsiflexion, both the normal and implanted ankles had similar sagittal midline talar contact positions but with plantar flexion, implanted ankles had increased posterior talar contact. Contact points on the distal tibia revealed that the lateral surface contacted at the midline or posterior throughout range-of-motion with minimal translation. The medial distal tibia contacted the talus posterior on plantarflexion and often moved anteriorly with dorsiflexion. This translation described relative external rotation of the distal tibia on plantar flexion and internal rotation on dorsiflexion. The measured distances were larger for the implanted ankles with higher variability. The average range-of-motion was 37.4 degrees for normal ankles and 32.3 degrees for implanted ankles. This study defines the normal kinematic rotational and translational motions of the ankle joint by accurately describing the three dimensional joint orientations. The implanted ankles experienced rotational and translational motions but had contacts more posterior, possibly related to surgical technique or alterations of ligamentous tension. PMID- 10808977 TI - Can tailored interventions increase mammography use among HMO women? AB - BACKGROUND: Telephone counseling and tailored print communications have emerged as promising methods for promoting mammography screening. However, there has been little research testing, within the same randomized field trial, of the efficacy of these two methods compared to a high-quality usual care system for enhancing screening. This study addressed the question: Compared to usual care, is tailored telephone counseling more effective than tailored print materials for promoting mammography screening? DESIGN: Three-year randomized field trial. PARTICIPANTS: One thousand ninety-nine women aged 50 and older recruited from a health maintenance organization in North Carolina. INTERVENTION: Women were randomized to 1 of 3 groups: (1) usual care, (2) tailored print communications, and (3) tailored telephone counseling. MAIN OUTCOME: Adherence to mammography screening based on self-reports obtained during 1995, 1996, and 1997. RESULTS: Compared to usual care alone, telephone counseling promoted a significantly higher proportion of women having mammograms on schedule (71% vs 61%) than did tailored print (67% vs 61%) but only after the first year of intervention (during 1996). Furthermore, compared to usual care, telephone counseling was more effective than tailored print materials at promoting being on schedule with screening during 1996 and 1997 among women who were off-schedule during the previous year. CONCLUSIONS: The effects of the intervention were most pronounced after the first intervention. Compared to usual care, telephone counseling seemed particularly effective at promoting change among nonadherent women, the group for whom the intervention was developed. These results suggest that telephone counseling, rather than tailored print, might be the preferred first-line intervention for getting nonadherent women on schedule for mammography screening. Many questions would have to be answered about why the tailored print intervention was not more powerful. Nevertheless, it is clear that additional interventions will be needed to maintain women's adherence to mammography. Medical Subject Headings (MeSH): mammography screening, telephone counseling, tailored print communications, barriers. PMID- 10808979 TI - Physical activity, food choice, and weight management goals and practices among US college students. AB - INTRODUCTION: Physical activity and a healthy diet have been recommended to help reverse the increasing prevalence of overweight among adolescents and adults in the United States. METHODS: Data is from the 1995 National College Health Risk Behavior Survey. A representative sample of US undergraduate college students (n = 4609) were analyzed to examine associations of physical activity and food choice with weight management goals and practices. RESULTS: Based on self reported height and weight, 35% of students were overweight or obese (body mass index > or = 25.0). Nearly half (46%) of all students reported they were trying to lose weight. Female students were less likely than male students to be overweight, but more likely to be trying to lose weight. Among female and male students, using logistic regression to control for demographics, trying to lose weight was associated with participation in vigorous physical activity and strengthening exercises, and consumption of < or = 2 servings/ day of high-fat foods. Female and male students who reported using exercise to lose weight or to keep from gaining weight were more likely than those who did not to participate in vigorous, strengthening, and moderate physical activity, and were more likely to eat > or = 5 servings/day of fruits and vegetables and < or = 2 servings/day of high-fat foods. Among students who were trying to lose weight, only 54% of females and 41% of males used both exercise and diet for weight control. CONCLUSION: Colleges should implement programs to increase student awareness of healthy weight management methods and the importance of physical activity combined with a healthy diet. PMID- 10808978 TI - Health care reform in Oregon: the impact of the Oregon Health Plan on utilization of mammography. AB - BACKGROUND: In 1994, Oregon implemented the Oregon Health Plan (OHP), extending health care coverage under a system of capitated managed care to uninsured citizens living below the Federal Poverty Level (FPL). We conducted a study to measure receipt of clinical preventive services among women newly enrolled in the OHP. METHODS: Six hundred and sixty six women aged 52-64, and living below the FPL in Oregon were randomly selected from OHP enrollment rosters and interviewed by telephone. A follow-up survey was conducted 1 year later. The main outcome of interest was receipt of a screening mammogram during the first year in the OHP. RESULTS: At enrollment 17% (65/383) of participants had never had health care coverage. Sixty-six percent of the women (220/333) were overdue for a mammogram. Fifty-five percent (121/220) reported cost as the main reason they had not had this procedure. Mammography rates doubled under the OHP (21% to 52%, 95% CI = 0.25-0.38, p < 0.001). Among women who were overdue for a mammogram at the time they enrolled, an expressed plan to get a mammogram (OR3.0, 95% CI = 1.1-8.7, p = 0.04), citing cost as the main reason for being overdue (OR3.0, 95% CI = 1.3-7.2, p = 0.014), receipt of a routine checkup (OR9.5, 95%CI = 3.7-24.9, p < 0.001), and health care provider's (HCP's) recommendation for mammography (OR8.1,95% CI = 2.9-23.0, p < 0.001) were independently associated with receipt of a mammogram. CONCLUSION: The OHP enrolled and successfully delivered clinical preventive services to a medically under served population. Even after removing the financial barrier, obstacles to mammography remain. These may be overcome by health systems changes to insure receipt of routine checkups and appropriate provider recommendations. PMID- 10808980 TI - Project GRAD: two-year outcomes of a randomized controlled physical activity intervention among young adults. Graduate Ready for Activity Daily. AB - INTRODUCTION: Project Graduate Ready for Activity Daily evaluated a program to promote physical activity through the transition of university graduation in a randomized controlled trial. METHODS: Three hundred thirty-eight university seniors participated in either a cognitive-behavioral intervention course or a knowledge-oriented general health course during the semester before graduation. Behaviorally oriented phone and mail follow-up was delivered to the intervention group for 18 months. Physical activity outcomes and mediating variables were assessed at baseline, 1 and 2 years (93% retention rate). RESULTS: There were no significant intervention effects on physical activity outcomes at 2 years for either men or women. Experiential and behavioral processes of change were significantly improved for intervention women over 2 years. CONCLUSIONS: Despite excellent participation in a theoretically based, well-attended intervention, few long-term effects on physical activity or its mediators were found. Additional research is needed to determine optimal interventions for physical activity and to validate or alter current behavior change theory. PMID- 10808981 TI - Seven-year tracking of dietary factors in young adults: the CARDIA study. AB - OBJECTIVES: This report determines the extent to which young adults in the highest and lowest intake quintiles of 13 nutrients remain in the same or adjacent quintiles (i.e., "tracked") relative to each other, over 7 years. METHODS: Data from baseline and year 7 of the CARDIA study were divided into race/gender-specific quintiles for each nutrient and cross-tabulated. RESULTS: For most nutrients, over 60% of those in the lowest absolute intake quintile at year 0 remained in the lowest or second-lowest quintile at year 7. A similar pattern was seen with highest absolute intake quintiles at years 0 and 7. Tracking was attenuated when nutrient density, rather than absolute intake, was examined. CONCLUSIONS: Ingrained dietary habits may cause high- or low-intake groups to retain relative ranking, even in the face of secular, age-, or lifestyle-related trends in dietary intake. PMID- 10808982 TI - Race/ethnicity, social class and their relation to physical inactivity during leisure time: results from the Third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Physical inactivity is more prevalent among racial and ethnic minorities than among Caucasians. It is not known if differences in participation in leisure time physical activity are due to differences in social class. Thus, this paper provides estimates of the prevalence of physical inactivity during leisure time and its relationship to race/ethnicity and social class. METHODS: This was a national representative cross-sectional survey with an in-person interview and medical examination. Between 1988 and 1994, 18,885 adults aged 20 or older responded to the household adult and family questionnaires as part of the Third National Health and Nutrition Examination Survey . Mexican-Americans and African-Americans were over-sampled to produce reliable estimates for these groups. Multiple assessment of social class included education, family income, occupation, poverty status, employment status, and marital status. RESULTS: The age-adjusted prevalence (per 100) of adults reporting leisure time inactivity is lower among Caucasians (18%) than among African-Americans (35%) and Mexican Americans (40%). African-American and Mexican-American men and women reported higher prevalence of leisure time inactivity than their Caucasian counterparts across almost every variable, including education, family income, occupation, employment, poverty and marital status. CONCLUSIONS: Current indicators of social class do not seem to explain the higher prevalence of physical inactivity during leisure time among African-American and Mexican-American. More research is needed to examine the effect of other constructs of social class such as acculturation, safety, social support and environmental barriers in promoting successful interventions to increase physical activity in these populations. PMID- 10808983 TI - Tobacco, alcohol, and caffeine use and cessation in early pregnancy. AB - OBJECTIVES: Little is known about what happens when individuals attempt to make multiple behavior changes simultaneously. Pregnant women in particular are often in the position of needing to change several behaviors at once, including giving up more than one pleasurable substance. We investigated the success of pregnant women in spontaneously quitting tobacco, alcohol, or caffeine, alone or in combination. METHODS: Pregnant women (n = 7489) were identified in the practices of large health maintenance organizations in Seattle and Minneapolis and were interviewed by telephone. Analyses examined the patterns of using and quitting more than one substance, and the extent to which using more than one substance predicts ability to quit other substances. RESULTS: Use of the three substances tended to cluster within individuals. Users of multiple substances were less likely to quit each substance than users of single substances. However, in the subgroup of multiple substance users who had quit one substance, having quit a second substance was more, rather than less, common. In multivariate analyses predicting quitting, demographic variables, and not having been pregnant previously were significant predictors of quitting each substance; being a nonsmoker predicted quitting alcohol, and being a nonsmoker and nondrinker predicted quitting caffeine. CONCLUSIONS: The reasons for difficulty in quitting more than one substance are unknown but may include the difficulty of formulating appropriate behavioral strategies or less concern about healthy behavior in pregnancy. Many women in the study successfully quit using two substances, however, and counseling should focus on achieving that outcome. PMID- 10808984 TI - Demographic predictors of cancer screening among Filipino and Korean immigrants in the United States. AB - BACKGROUND: Little is known about cancer-screening practices of various Asian subgroups, and even less is known about factors that may predict screening in these populations. DESIGN: Two independent surveys were conducted with 218 Filipino and 229 Korean female immigrants, aged 50 years and older, residing in Los Angeles. RESULTS: In these convenience samples, 48% of Filipino and 41% of Korean women reported receipt of a Pap smear within the past 2 years; 41% of Filipino and 25% of Korean women reported receipt of a mammogram and a clinical breast exam within the past 2 years; and 25% of Filipino and 38% of Korean women reported colorectal cancer screening (blood stool test within the past 12 months or sigmoidoscopy/colonoscopy within the past 5 years). Only 14% of Filipino and 10% of Korean women were adherent to cancer-screening guidelines for all three sites. These differences in screening rates were statistically significant in multivariate analyses of the combined sample, controlling for all demographic characteristics, including age, percent of lifetime in the United States, education, marital status, health insurance, employment, and ethnicity. The two variables that were most consistently independently associated with adherence to cancer screening in both samples were higher percentage of lifetime spent in the United States and ever having had a checkup when no symptoms were present. CONCLUSIONS: These two variables-percent of lifetime in the United States and ever having had a checkup when no symptoms were present-can alert a physician that cancer-screening tests may be overdue among Korean and Filipino immigrants in the United States. Future research should identify predictors of cancer screening among other Asian immigrant groups and U.S.-born Asian women to assist in targeting intervention efforts. PMID- 10808985 TI - Interactive computer technology for skin cancer prevention targeting children. AB - BACKGROUND: Computer technology has become an integral part of health care, yet there have been few studies exploring the use of multimedia technology in the prevention of cancer, especially targeting children. OBJECTIVE: The aims of this study were to develop and evaluate a new multimedia computer program for the primary prevention of skin cancer among a childhood population. DESIGN AND PARTICIPANTS: An interactive CD-ROM program was developed, then pilot tested in a public elementary school in rural North Carolina. This intervention trial involved 8 third- and fourth-grade classes (N = 209 students), randomized into 3 groups: computer intervention, standard teacher-led intervention, and controls. MAIN OUTCOME MEASURES: Students were tested using pre- and postintervention surveys that measured knowledge, attitudes, and self-reported behaviors. A 7 month follow-up survey was performed. RESULTS: There was a significant increase in postintervention knowledge for the computer group when compared to either the teacher-led or control groups (mean scores out of 100: 75.2, 59.5, 55.0, respectively; p < 0.001). Attitudes about suntanning demonstrated a significant difference between the 3 groups (mean scores out of 100: 64.0, 53.0, 48.6, respectively; p = 0.002). There were slight improvements in the behavioral scores, especially among the computer group, but the overall differences were not significant. Similar overall results were found for the long-term follow-up survey, except that attitudes about suntanning no longer demonstrated a significant difference. CONCLUSION: These results indicate that this new educational tool is an effective way to introduce health education programs for young children in typical classroom settings. This prototype may serve as a model for the development of future preventive school-based programs, including applications to other conditions associated with high-risk behaviors among children. PMID- 10808986 TI - Gender differences in quality-adjusted survival using a Health-Utilities Index. AB - PURPOSE: Women live longer than men but experience high morbidity during later years. We attempt to represent life expectancy with adjustments for quality of life for men and women in the United States. DATA SOURCES: Survival estimates were obtained from Vital Statistics of the United States Life Tables. Quality-of life data were obtained for 12,220 participants, aged 32-85 years, in the 1982 1984 National Health and Nutrition Examination Survey I Epidemiologic Follow-Up Study (NHEFS). METHOD: Using public data tapes, scores for the Health-Utilities Index (HUI) were imputed for NHEFS. These scores were calculated separately for men and women in the United States population and broken down by age. Using mortality data, the quality-adjusted life expectancy was calculated separately for men and women. RESULTS: The current life expectancy among men aged 32 years was 39.45 years. For women aged 32 years it was 44.83 years, suggesting a 5.38 female life-expectancy advantage. The life expectancy, adjusted for quality of life, was 31.8 years for men versus 33.1 years for women. Adjustment for quality of life reduced the 5.38-year female advantage to 1.3 years. CONCLUSIONS: Although women enjoy longer life expectancies than do men, this advantage is reduced when quality adjustments are used. The finding reflects high levels of morbidity among older women. PMID- 10808987 TI - The quantity and quality of physical activity among those trying to lose weight. AB - BACKGROUND: Regular exercise to elicit caloric expenditure is an important component for achieving weight loss. The Healthy People 2000 objectives recommend regular sustained physical activity lasting 30 minutes, five days per week (Objective 1.3) particularly for weight loss. Moreover, this recommendation has been restated for weight loss and overall health benefits in the Centers for Disease Control and Prevention / American College of Sports Medicine (CDC/ACSM) statement and Surgeon General's Report (SGR) on Physical ActivitY and Health. Thus, we sought to identify the relative quality, and quantity of physical activity among people trying to lose weight. DESIGN: Cross-sectional self reported data from the West Virginia Behavioral Risk Factor Surveillance System (BRFSS) were used. The BRFSS is a state-based telephone survey of adults that uses a multistage cluster design based on the Waksberg method of random-digit dialing. Data froM 2769 men and 4490 women were obtained from the 1992, 1994, and 1996 surveys. RESULTS: Half (49.6%) of individuals trying to lose weight did not engage in any physical activity. Further, only 15% of respondents trying to lose weight reported exercising regularly. Nevertheless, those trying to lose weight were more likely (OR [odds ratio] = 1.3; 95% CI [confidence interval], 1.14, 1.51, p < 0.001) to exercise regularly than those not trying to lose weight. In particular, women trying to lose weight were significantly more likely (OR = 1.45; 95% CI, 1.22,1.74, p < 0.001) to exercise regularly than women not trying to lose weight. Conversely, men trying to lose weight were no more likely to exercise regularly (p = .23) than men not trying to lose weight. Among respondents who were using exercise for weight loss, only 14.7% were expending > or =1000 kcal/week and 18.2% were expending > or =500 kcal/week. Weekly expenditure rates of > or =1000 kcal/week were more likely to occur among men (17%) than women (13.8%), in younger age groups, and among those with higher educational attainment. CONCLUSION: These data suggest that while certain individuals trying to lose weight are more likely to engage in regular physical activity, most persons trying to lose weight have not adopted regular physical activity as part of their weight loss practice. These results suggest that public health efforts to effectively integrate physical activity into weight control practices of West Virginians have been minimally successful. PMID- 10808988 TI - A threat to the public health workforce: evidence from trends in preventive medicine certification and training. AB - Evidence of a growing need for preventive medicine specialists is the congruence between needed competencies for practice in the current health care environment, as identified by the Council on Graduate Medical Education (COGME) and in other national reports, and the core competencies of preventive medicine residents. The total number of certified specialists in preventive medicine is 6091. The proportion of self-designated preventive medicine specialists among all U.S. physicians is on the decline and the greatest decline has been among those in public health (PH) and general preventive medicine (GPM). In addition, the total number of preventive medicine residents is on the decline, and the decline has been greatest among those training in PH and combined PH/GPM. One of the reasons for this decline has been inadequate funding due to the absence of Medicare graduate medical education (GME) financing for population-based vs. individual patient care services and meager and diminishing Title VII support. A paucity of faculty is apparent in medical schools with residency training and board certification in preventive medicine. Several actions may help reverse this trend and assure adequate numbers of preventive medicine specialists: expansion of Title VII to increase the number of residents receiving stipends and tuition, adding infrastructure support for faculty development and funding of demonstration projects in distance learning and in joint generalist/ preventive medicine residency training. Medicare GME reform should include recognition of population-based services and inclusion of preventive medicine residencies in provisions for "nonhospital-based" training and in up-weighting methodologies for primary care training. Expansion of Veterans Affairs, National Institute for Occupational Safety and Health, and Department of Defense support is also needed as is attention to resident debt reduction. PMID- 10808989 TI - Preventive medicine in a new millennium. PMID- 10808990 TI - From public health and medicine to population health improvement: the 50-year search for training in preventive medicine. PMID- 10808991 TI - Response to the report of the AAMC population health perspective panel. PMID- 10808992 TI - Herbicide movement and dissipation at four Midwestern sites. AB - This study was conducted to evaluate atrazine (2-chloro-4-ethylamino-6-isopropyl 1,3,5-triazine) and alachlor (2-chloro-N-(methoxymethyl)acetamide) dissipation and movement to shallow aquifers across the Northern Sand Plains region of the United States. Sites were located at Minnesota on a Zimmerman fine sand, North Dakota on Hecla sandy loam, South Dakota on a Brandt silty clay loam, and Wisconsin on a Sparta sand. Herbicide concentrations were determined in soil samples taken to 90 cm four times during the growing season and water samples taken from the top one m of aquifer at least once every three months. Herbicides were detected to a depth of 30 cm in Sparta sand and 90 cm in all other soils. Some aquifer samples from each site contained atrazine with the highest concentration in the aquifer beneath the Sparta sand (1.28 microg L(-1)). Alachlor was detected only once in the aquifer at the SD site. The time to 50% atrazine dissipation (DT50) in the top 15 cm of soil averaged about 21 d in Sparta and Zimmerman sands and more than 45 d for Brandt and Hecla soils. Atrazine DT50 was correlated positively with % clay and organic carbon (OC), and negatively with % fine sand. Alachlor DT50 ranged from 12 to 32 d for Zimmerman and Brandt soils, respectively, and was correlated negatively with % clay and OC and positively with % sand. PMID- 10808993 TI - Fenuron sorption on homoionic natural and modified smectites. AB - The adsorption isotherms of fenuron (1,1-dimethyl-3-phenylurea) on three smectites (SWy and SAz montmorillonites and SH hectorite) differing in their layer charge (SH 400/microl and plasma HIV RNA copies > 5000/ml) were randomly assigned to receive highly active antiretroviral therapy (HAART) alone or HAART plus rIL-2. Plasma viremia (measured by a commercial RT-PCR assay) and the number of provirus-infected cells (measured by an endpoint cell dilution PCR assay) were monitored at the enrollment and after 12 weeks of treatment. The results indicate that while HAART and HAART plus rIL-2 are both able to significantly reduce plasma viremia after 12 weeks of treatment, a significant reduction of the number of provirus-infected cells can be achieved only by treatment with HAART plus rIL-2. PMID- 10809020 TI - Comparison of human cytomegalovirus DNA polymerase activity for ganciclovir resistant and -sensitive clinical strains. AB - Previously, we found that a ganciclovir (GCV)-resistant clinical human cytomegalovirus (HCMV) isolate had an amino acid substitution at codon 501 (Leu - > Phe) in the delta-region C of the DNA polymerase gene. DNA polymerases have now been (partially) purified from both the GCV-resistant and sensitive parental strains and the activity of DNA polymerase and 3'-5' exonuclease compared. With respect to DNA polymerase activity, the Michaelis constant (Km) and maximum velocity (Vmax) of the GCV-resistant strain for the DNA template were lower than those of the GCV-sensitive strain. With respect to 3'-5' exonuclease activity, the Km and Vmax of the GCV-resistant strain for the DNA substrate in the presence of ammonium sulfate were lower than those of the GCV-sensitive strain, while being similar in the absence of ammonium sulfate. Although the polymerase activity of the two strains showed almost the same sensitivity for the different polymerase inhibitors, the 3'-5' exonuclease activity of the GCV-resistant strain was more resistant to these inhibitors than that of the GCV-sensitive strain. PMID- 10809019 TI - Quantification of the impact of HIV-1 reverse transcriptase and protease mutations on the efficacy of rescue HAART. AB - The reduction in the efficacy of rescue treatment (administered on a clinical basis) due to drug resistance was retrospectively quantified in 55 human immunodeficiency virus type 1 (HIV-1)-infected patients failing highly active antiretroviral therapy (HAART) by using a novel score calculation system based upon HIV-1 reverse transcriptase (RT) and protease (PR) mutations. Patients were all naive for nelfinavir (NFV) and efavirenz (EFV) and were assigned to one of the following rescue therapy schedules: (i) 17 patients received NFV + EFV + stavudine (d4T) (group A); (ii) 14 patients received NFV + saquinavir (SQV) + lamivudine (3TC) + d4T/zidovudine (AZT) (group B); (iii) 19 patients received NFV + d4T + didanosine (ddI)/3TC/zalcitabine (ddC) (group C); (iv) five patients received miscellaneous treatments including NFV (group D). Responders were considered patients showing a drop in HIV-1 RNA level > 0.5 log10 after 3 months of therapy. Forty-eight (28 responders and 20 non-responders) out of 55 patients completed the first 3 months of rescue therapy and reduction in HIV-1 viral load was found to be significantly higher in group A compared to groups B and C (81.2% responders vs. 38.5 and 40.0%, respectively). At baseline, no patient carried EFV or d4T-resistant HIV-1 strains, despite prolonged administration of d4T, while 41/48 (87.2%) patients had mutations conferring resistance to NFV in the absence of previous treatment with this drug. A significant inverse correlation between reduction in viral load and reduction in therapy efficacy due to drug resistance, as determined by the score calculation system, was found (r = 0.62). A cut-off value of 36% reduction in therapy efficacy showed a positive predictive value (capacity to detect failure of rescue treatment) of 81.2% and a negative predictive value (ability to detect successful treatment) of 75.8%. In addition, 45 out of 48 patients completed also the 9-12 month period of rescue therapy and 10/28 responders had a rebound in HIV-1 viral load level detected after the first 3 months of rescue therapy. Of these, 5/7 (71.4%) showed a further reduction in rescue therapy efficacy due to the emergence of new mutations. PMID- 10809021 TI - 1,3-Dihydroxyacridone derivatives as inhibitors of herpes virus replication. AB - The nuclear enzyme DNA topoisomerase II is a candidate pharmacological target for treating herpes virus infections and the novel catalytic inhibitors, 7-chloro-1,3 dihydroxyacridone (compound 1), and 1,3,7-trihydroxyacridone (2) are potential lead compounds [Bastow, K.F., Itoigawa, M., Furukawa, H., Kashiwada, Y., Bori, I.D., Ballas, L.M., Lee, K.-H., 1994. Antiproliferative actions of 7-substituted 1,3-dihydroxyacridones; possible involvement of DNA topoisomerase II and protein kinase C as biochemical targets. Bioorg. Med. Chem. 2, 1403-1411; Vance, J.R., Bastow, K.F., 1999. Inhibition of DNA topoisomerase II catalytic activity by the antiviral agents 7-chloro,1,3-dihydroxyacridone and 1,3,7-trihydroxyacridone. Biochem. Pharmacol. 58, 703-708]. In this report, four new 1,3-dihydroxyacridone analogs with functional groups at either the 5-, 6- or 8-positions (compounds 3 6) were synthesized. Target compounds, three other analogs including compounds 1 and 2 and three anticancer drugs that inhibit DNA topoisomerase II (etoposide, amsacrine and aclarubicin) were then evaluated as selective inhibitors of herpes simplex virus (HSV) replication in cell culture and as enzyme inhibitors in vitro. Etoposide and amsacrine inhibited HSV but acted non-selectively. In general, the activities of 1,3-dihydroxyacridone derivatives as selective anti HSV agents and as enzyme inhibitors varied inversely suggesting that DNA topoisomerase II probably is not the critical antiviral target. The 5-Cl congener (compound 3) was the most selective agent (about 26-fold under a stringent assay condition) but was not an enzyme inhibitor. Results of exploratory mechanistic studies with compounds 1 and 3 show that HSV replication was blocked at a stage after DNA and late protein synthesis. The acridone derivatives were also tested against human cytomegalovirus (HCMV) replication but none of them were active. PMID- 10809022 TI - Treatment of lethal Ebola virus infection in mice with a single dose of an S adenosyl-L-homocysteine hydrolase inhibitor. AB - Ebola Zaire virus causes lethal hemorrhagic fever in humans, for which there is no effective treatment. A variety of adenosine analogues inhibit the replication of Ebola virus in vitro, probably by blocking the cellular enzyme, S-adenosyl-L homocysteine hydrolase, thereby indirectly limiting methylation of the 5' cap of viral messenger RNA. We previously observed that adult, immunocompetent mice treated thrice daily for 9 days with 2.2-20 mg/kg of an adenosine analogue, carbocyclic 3-deazaadenosine, were protected against lethal Ebola virus challenge. We now report that a single inoculation of 80 mg/kg or less of the same substance, or of 1 mg/kg or less of another analogue, 3-deazaneplanocin A, provides equal or better protection, without causing acute toxicity. One dose of drug given on the first or second day after virus infection reduced peak viremia more than 1000-fold, compared with mock-treated controls, and resulted in survival of most or all animals. Therapy was less effective when administered on the day of challenge, or on the third day postinfection. Single or multiple doses of the same medications suppressed Ebola replication in severe combined immunodeficient mice, but even daily treatment for 15 consecutive days did not eliminate the infection. PMID- 10809023 TI - A novel approach using an attenuated recombinant vaccinia virus to test the antipoxviral effects of handsoaps. AB - Evidence indicates an increase in nosocomial and household infections due to viruses (Jeffries, D.J., 1995. Viral hazards to and from health care workers. J. Hosp. Infect. 30, 140-155). An antiviral assay was developed for evaluating efficiency of handsoaps at inactivating cell-free and cell-associated virus. A recombinant vaccinia virus, lacking a virulence factor (Isaacs, S.N., Kotwal, G.J., Moss, B., 1992. Vaccinia virus complement-control protein prevents antibody dependent complement-enhanced neutralization of infectivity and contributes to virulence. Proc. Natl. Acad. Sci. USA 89, 628-632), whose construction was described earlier (Kotwal, G.J., Isaacs, S.N., McKenzie, R., Frank, M.M., Moss, B., 1990. Inhibition of the complement cascade by the major secretory protein of vaccinia virus. Science 250, 827-830), was used as the representative poxvirus. Two antibacterial handsoaps, one surgical handsoap, one moisturizing handsoap, and a sanitizing agent were tested. An aliquot of the virus was mixed and incubated with soap, then titrated onto BSC-1 cells for incubation at 37 degrees C for 48 h. The soaps' effect on cell-associated virus was tested similarly. The antibacterial soaps inactivated all cell-free virus in 1 min. The surgical soap was effective with a 5-min incubation. None of the soaps eliminated all of the cell-associated virus in 1 min. This safe and reproducible assay seems efficient to establish the comparative efficacy of household and surgical soaps. PMID- 10809024 TI - Links between Helicobacter pylori infection, cobalamin deficiency, and pernicious anemia. PMID- 10809025 TI - Productivity in primary care: work smarter, not harder. PMID- 10809026 TI - Beta-blockers: the new standard of therapy for mild heart failure. AB - Many physicians are reluctant to prescribe beta-blockers to patients with mild heart failure, especially when standard therapy (diuretics and an angiotensin converting enzyme inhibitor, with or without digitalis glycosides) seems to be effective at relieving symptoms. However, current first-line medications for heart failure either ignore or incompletely inhibit adrenergic activation, one of the primary contributors to progressive left ventricular systolic dysfunction. Thus, even effective standard "triple" therapy does not safeguard the patient against further catastrophic deterioration of cardiac performance. Clinical trials have shown that the use of beta-blockers in addition to standard therapy improves left ventricular function, reduces hospitalizations, and-in the cases of bisoprolol, long-acting metoprolol, and carvedilol-improves survival in patients with chronic heart failure. In addition, carvedilol has been found to significantly slow disease progression even in mildly symptomatic patients. Though achieving beta-blockade in patients with heart failure requires extra effort by the clinician (appropriate patient selection, optimization of background therapy, initiating drug treatment at low doses, and titrating slowly with careful vigilance for early signs of clinical instability), the cost is small compared with the consequence of postponing adrenergic intervention. The educational objective of this article is to provide the primary care physician with a review of the current understanding of the pathophysiological characteristics underlying chronic systolic heart failure, the clinical benefits of administering beta-blockers during the early stages of heart failure, and the practical considerations of initiating therapy. PMID- 10809027 TI - Home blood pressure monitoring. AB - Hypertension is estimated to affect 43 to 56 million adults or 24% to 31% of the US population and is emerging as a major health problem in some countries in the Third World. Hypertension contributes to all the major atherosclerotic cardiovascular disease outcomes. PMID- 10809028 TI - Depression as an antecedent to heart disease among women and men in the NHANES I study. National Health and Nutrition Examination Survey. AB - BACKGROUND: Depression predicts morbidity and mortality among individuals who have coronary heart disease (CHD), and there is increasing evidence that depression may also act as an antecedent to CHD. The studies that have reported a relationship between depression and CHD incidence or mortality either were restricted to men only or analyzed women and men together. The present investigation was conducted to evaluate the differential effect depression may have on CHD incidence and mortality in women and men. RESEARCH METHODS: We analyzed data from 5007 women and 2886 men enrolled in the first National Health and Nutrition Examination Survey (NHANES I) who were free of CHD at the 1982-1984 interview and who had completed the Center for Epidemiologic Studies Depression Scale (CES-D). Participants were evaluated from the 1982 interview date either until the end of the study (1992 interview date) or until the occurrence of a CHD event. Using CHD incidence and CHD mortality (International Classification of Disease, Ninth Revision, codes 410-414) as the outcome variables, Cox proportional hazards regression models were developed to evaluate the relative risk (RR) of CHD incidence and mortality in the depressed women and men separately, controlling for standard CHD risk factors. RESULTS: The women experienced 187 nonfatal and 137 fatal events, compared with 187 nonfatal and 129 fatal events among the men. The adjusted RR of CHD incidence among depressed women was 1.73 (95% confidence internal [CI], 1.11-2.68) compared with nondepressed women. Depression had no effect on CHD mortality in the women (RR, 0.74; 95% CI, 0.40-1.48). The adjusted RR of CHD incidence among depressed men was 1.71 (95% CI, 1.14-2.56) compared with nondepressed men. Depressed men also had an increased risk of CHD mortality compared with their nondepressed counterparts, with an adjusted RR of 2.34 (95% CI, 1.54-3.56). CONCLUSIONS: In this sample, while controlling for possible confounding factors, depression was associated with an increased risk of CHD incidence in both men and women, as well as CHD mortality in men. Depression had no effect on CHD mortality in women. PMID- 10809029 TI - Health insurance and access to care for symptomatic conditions. AB - BACKGROUND: The uninsured receive less medical care than the insured. However, it is not known whether the uninsured are less likely to seek medical care for highly "serious" or "morbid" symptoms. METHODS: Participants in the National Access to Care Survey were asked whether they had experienced any of 15 highly serious or morbid symptoms. Those who did were asked whether they received medical care and, if care was not received, whether care was thought to have been necessary. RESULTS: A total of 574 respondents (16.4%) reported 794 new serious or morbid symptoms. Of these, 499 people (86.9%) had health insurance, and they reported 691 new symptoms; 75 (13.1%) lacked health insurance, and they reported 103 symptoms. The uninsured were less likely to have received medical care and more likely to say they did not receive care even though they thought it was needed (P = .001). Medical care was received for 45.4% of symptoms for the insured and 24.3% for the uninsured; care was not thought to have been needed for 41.0% of symptoms for the insured and 45.6% for the uninsured; and care was thought necessary but was not received for 13.6% of symptoms for the insured and 30.1% for the uninsured. In multivariate analysis, the uninsured were much less likely to have received care than the insured (adjusted odds ratio, 0.43; 95% confidence interval, 0.23-0.81). CONCLUSION: Lack of health insurance is a major barrier to receiving medical care, even for highly serious and morbid symptoms. PMID- 10809030 TI - Cost-effectiveness of the lower treatment goal (of JNC VI) for diabetic hypertensive patients. Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. AB - BACKGROUND: The recommendation of the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) to lower blood pressure (BP) in diabetic patients to less than 130/85 mm Hg may have negative economic consequences. A formal cost-effectiveness analysis was therefore performed, comparing the costs and potential benefits of a BP goal of less than 140/90 mm Hg (as recommended by JNC V) vs less than 130/85 mm Hg (as inJNC VI). METHODS: A 24-cell computer model was populated with costs (1996 dollars), relative risks, and age-specific base-line rates for death and 4 nonfatal adverse events (stroke, myocardial infarction, heart failure, and end stage renal disease), derived from published data. Costs and benefits were discounted at 3%. RESULTS: For 60-year-old diabetic persons with hypertension, treating to the lower BP goal increases life expectancy by 0.48 (discounted) years and lowers (discounted) lifetime medical costs by $1450 compared with treating BP to less than 140/90 mm Hg. The lower treatment BP goal results in an overall cost savings over a wide range of initial conditions, and for nearly all analyses for patients older than 60 years. CONCLUSIONS: Any incremental treatment for 60-year-olds that costs less than $414 annually and successfully lowers BP from below 140/90 to below 130/85 mm Hg would be cost saving in the long term, due to the reduction in attendant costs of future morbidity. The lower treatment goal recommended for high-risk hypertensive patients compares favorably in cost effectiveness with many other frequently recommended treatment strategies, and saves money overall for patients aged 60 years and older. PMID- 10809031 TI - Helicobacter pylori-related disease: guidelines for testing and treatment. AB - OBJECTIVE: To develop practical guidelines for the treatment of patients with suspected and documented Helicobacter pylori-related gastroduodenal diseases. METHODS: A panel of physicians with expertise in H. pylori reviewed, critically appraised, and synthesized the literature on assigned topics and presented their overviews to the panel. Consensus was obtained in controversial areas through discussion. RESULTS AND CONCLUSIONS: The panel recommended testing for H. pylori in patients with active ulcers, a history of ulcers, or gastric mucosa-associated lymphoid tissue lymphomas. Young, otherwise healthy patients with ulcerlike dyspepsia and those with a family history or fear of gastric cancer may also undergo H pylori testing. Non-endoscopic methods are preferred for H. pylori diagnosis. Dual medication regimens should not be used for therapy; twice-daily triple therapy with a proton pump inhibitor or ranitidine bismuth citrate, clarithromycin, and amoxicillin for 10 to 14 days is an appropriate therapy. Posttreatment assessment of H. pylori status using urea breath testing should be considered in patients with a documented history of ulcer disease or with persistent symptoms. PMID- 10809033 TI - The treatment of elderly diabetic patients with acute myocardial infarction: insight from Michigan's Cooperative Cardiovascular Project. AB - BACKGROUND: Diabetic patients with acute myocardial infarction (AMI) have higher morbidity and mortality rates than nondiabetic patients with AMI. Thus, reliable adherence to quality care is necessary in these patients to improve outcomes. We analyzed data from the Health Care Financing Administration's Cooperative Cardiovascular Project (CCP) in Michigan, addressing quality of care in diabetic patients with AMI. METHOD: All acute-care hospitals in Michigan had 8 consecutive months of baseline CCP data abstracted from medical records of all Medicare patients who were discharged with a principal diagnosis of AMI. Owing to the staggered 8-month periods, abstraction occurred for patients who were discharged between April 1, 1994, and July 31, 1995. RESULTS: Diabetic patients accounted for 33% of 8455 patients with AMI. Diabetic patients were primarily younger, female, and nonwhite. They had a greater frequency of non-Q-wave AMI and presented less often within 6 hours of their infarction. Comorbid conditions, such as hypertension, prior AMI, prior stroke, and/or prior revascularization, were more frequent in diabetic than in nondiabetic patients. Congestive heart failure occurred more frequently in diabetic patients. Length of stay (7.9 vs 7.0 days; P<.001), in-hospital mortality rates (16% vs 13%; P<.001), and rates for mortality within 30 days (21% vs 17%; P<.001) were higher in diabetic patients. CONCLUSIONS: Despite greater frequencies of comorbid conditions, poorer outcomes, and greater resource use, there is poor overall adherence to most quality indicators in diabetic patients with AMI. Better methods for systematizing proven prevention and treatment strategies in the care of patients with AMI are needed in this unique high-risk cohort. PMID- 10809032 TI - Azithromycin vs cefuroxime plus erythromycin for empirical treatment of community acquired pneumonia in hospitalized patients: a prospective, randomized, multicenter trial. AB - OBJECTIVE: To compare the efficacy and safety of azithromycin dihydrate monotherapy with those of a combination of cefuroxime axetil plus erythromycin as empirical therapy for community-acquired pneumonia in hospitalized patients. METHODS: Patients were enrolled in a prospective, randomized, multicenter study. The standard therapy of cefuroxime plus erythromycin was consistent with the American Thoracic Society, Canadian Community-Acquired Pneumonia Consensus Group, and Infectious Disease Society of America consensus guidelines. The doses were intravenous azithromycin (500 mg once daily) followed by oral azithromycin (500 mg once daily), intravenous cefuroxime (750 mg every 8 hours), followed by oral cefuroxime axetil (500 mg twice daily), and erythromycin (500-1000 mg) intravenously or orally every 6 hours. Randomization was stratified by severity of illness and age. Patients who were immunosuppressed or residing in nursing homes were excluded. RESULTS: Data from 145 patients (67 received azithromycin and 78 received cefuroxime plus erythromycin) were evaluable. Streptococcus pneumoniae and Haemophilus influenzae were isolated in 19% (28/145) and 13% (19/145), respectively. The atypical pathogens accounted for 33% (48/145) of the etiologic diagnoses; Legionella pneumophila, Chlamydia pneumoniae, and Mycoplasma pneumoniae were identified in 14% (20/ 145), 10% (15/145), and 9% (13/145), respectively. Clinical cure was achieved in 91% (61/67) of the patients in the azithromycin group and 91% (71/78) in the cefuroxime plus erythromycin group. Adverse events (intravenous catheter site reactions, gastrointestinal tract disturbances) were significantly more common in patients who received cefuroxime plus erythromycin (49% [30/78]) than in patients who received azithromycin (12% [8/67]) (P<.001). CONCLUSIONS: Treatment with azithromycin was as effective as cefuroxime plus erythromycin in the empirical management of community-acquired pneumonia in immunocompetent patients who were hospitalized. Azithromycin was well tolerated. PMID- 10809034 TI - Epidemiology of de novo acute renal failure in hospitalized African Americans: comparing community-acquired vs hospital-acquired disease. AB - BACKGROUND: The high incidence and prevalence of end-stage kidney disease among African Americans is well known, but the epidemiology of acute renal failure (ARF) among African Americans is unknown. This study was designed to determine the incidence, associated risk factors, and prognosis of ARF in hospitalized African Americans and to compare these variables in hospital-acquired ARF (HA ARF) against community-acquired ARF (CA-ARF). METHODS: A 3-year (1994-1996), computer-assisted retrospective analysis of hospital discharges with confirmed diagnoses of ARF. One hundred of 240 cases met the inclusion criteria for de novo ARF. Demographic, laboratory, and clinical profiles of all patients were retrieved and subdivided into CA-ARF and HA-ARF. Both analysis of variance and chi2 tests were used for analysis. Survival regression used both the Cox proportional hazards and Kaplan-Meier models. RESULTS: The incidence of CA-ARF was 3.5 times greater than that of HA-ARF (0.55% vs 0.15%). The mean age of all patients was 54 years with a 67% male predominance. There were no significant differences in age, sex, peak serum creatinine levels, or underlying medical history. Prerenal causes of ARF were more common among CA-ARF than HA-ARF (35% vs 19%; P = .07), but intra-renal causes were more common among HA-ARF (81% vs 55%; P = .07). All cases of obstruction occurred in CA-ARF. Mortality was higher in HA ARF (59% vs 33%; P = .03), and the incidence of recognized predictors of mortality was higher in patients with HA-ARF than in those with CA-ARF: oliguria (59% vs 35%; P = .04); sepsis (73% vs 35%; P = .004); stay in the intensive care unit (ICU) or mechanical ventilation (55% vs 6%; P<.001); and multiorgan failure (59% vs 24%; P = .002). Those with HA-ARF were twice as likely to require dialysis as those with CA-ARF. The mortality was high in younger patients with CA ARF and in older patients with HA-ARF, but the dialysis-related mortality rate was 3-fold higher among patients with HA-ARF. While mean +/- SD length of hospital stay was more prolonged in HA-ARF than CA-ARF (26 +/- 28 days vs 12 +/- 11 days; P<.001), the 120-day survival rate was lower in HA-ARF than CA-ARF (43% vs 66%; P = .05). The HA-ARF status was associated with a relative risk of 2.5 (confidence interval, 1.1-5.5; P = .03) for shortened survival. CONCLUSIONS: The overall epidemiologic characteristics of ARF among hospitalized African Americans seem to be comparable to those in whites, but the difference in incidence between CA-ARF and HA-ARF was much higher in African Americans. In view of the high mortality and morbidity rates associated with ARF and the fact that younger African American patients with CA-ARF were more likely to die than their older counterparts, we recommend that renal failure awareness be incorporated into community-based health educational programs in African American populations. PMID- 10809035 TI - Earlier initiation of antibiotic treatment for severe infections after interventions to improve the organization and specific guidelines in the emergency department. AB - OBJECTIVE: To examine whether combined interventions improve the timely administration of antibiotic therapy and acquisition of material for culture from patients admitted to the emergency department with a serious infectious disease. METHODS: Guidelines and educational programs were developed to facilitate timely antibiotic administration: guidelines on handling patients with serious infections and on ordering immediate treatment, guidelines on obtaining culture samples, lectures to medical and nursing staff, improvement of availability of antibiotics in the emergency department, and removal of financial restraints on stocking and ordering of antibiotics. Fifty consecutive patients were evaluated after this series of interventions and compared with the results in 50 patients evaluated before the interventions. The interval from presentation to the emergency department until the administration of antibiotics, number of samples taken for microbiological investigations, and number of patients receiving a first dose of antibiotic at routinely scheduled drug distribution rounds were evaluated. RESULTS: The median time to the initial dose of antibiotics administered decreased from 5.0 hours to 3.2 hours (P = .04). The number of blood cultures obtained did not change. The percentage of sputum cultures obtained increased from 28% to 50%, and the percentage of urine cultures obtained increased from 50% to 100%. The percentage of patients whose first dose of antibiotic was delayed until a routinely scheduled drug distribution round decreased from 54% to 32% (P = .03). CONCLUSIONS: Combined interventions to expedite diagnostic and therapeutic actions through directed clinical practice guidelines and organizational measures are successful. This may lead to a substantial quality improvement in the process of care. PMID- 10809036 TI - Effects of weight loss with orlistat on glucose tolerance and progression to type 2 diabetes in obese adults. AB - BACKGROUND: Orlistat is a gastrointestinal lipase inhibitor that reduces dietary fat absorption by approximately 30%, promotes weight loss, and may reduce the risk of developing impaired glucose tolerance and type 2 diabetes in obese subjects. OBJECTIVE: To test the hypothesis that orlistat combined with dietary intervention improves glucose tolerance status and prevents worsening of diabetes status more effectively than placebo. METHODS: We pooled data from 675 obese (body mass index, 30-43 kg/m2) adults at 39 US and European research centers in 3 randomized, double-blind, placebo-controlled multicenter clinical trials. Subjects received placebo plus a low-energy diet during a 4-week lead-in period. On study day 1, the diet was continued, and subjects were randomized to receive placebo 3 times a day (n=316) or treatment with orlistat, 120 mg 3 times a day (n=359), for 104 weeks. A standard 3-hour oral glucose tolerance test was performed on day 1 and at the end of treatment. MAIN OUTCOME MEASURES: The categorical assessment of glucose tolerance status (normal, impaired, diabetic) and changes in status from randomization to end of treatment were the primary efficacy measures. The secondary measures were fasting and postchallenge glucose and insulin levels. RESULTS: The mean length of follow-up was 582 days. Subjects who were treated with orlistat lost more weight (mean +/- SEM, 6.72 +/- 0.41 kg from initial weight) than subjects who received placebo (3.79+/-0.38 kg; P<.001). A smaller percentage of subjects with impaired glucose tolerance at baseline progressed to diabetic status in the orlistat (3.0%) vs placebo (7.6%) group. Conversely, among subjects with impaired glucose tolerance at baseline, glucose levels normalized in more subjects after orlistat treatment (71.6%) vs placebo (49.1%; P=.04). CONCLUSIONS: The addition of orlistat to a conventional weight loss regimen significantly improved oral glucose tolerance and diminished the rate of progression to the development of impaired glucose tolerance and type 2 diabetes. PMID- 10809037 TI - Understanding racial variation in the use of coronary revascularization procedures: the role of clinical factors. AB - BACKGROUND: Black patients undergo coronary artery bypass grafting and percutaneous transluminal coronary angioplasty less often than white patients. It is unclear how racial differences in clinical factors contribute to this variation. METHODS: A retrospective cohort study was performed of 666 male patients (326 blacks and 340 whites), admitted to 1 of 6 Veterans Affairs hospitals from October 1, 1989, to September 30, 1995, with acute myocardial infarction or unstable angina who underwent cardiac catheterization. The primary comparison was whether racial differences in percutaneous transluminal coronary angioplasty and coronary artery bypass grafting rates persisted after stratifying by clinical appropriateness of the procedure, measured by the appropriateness scale developed by the RAND Corporation, Santa Monica, Calif. RESULTS: Whites more often than blacks underwent a revascularization procedure (47% vs 28%). There was substantial variation in black-white odds ratios within different appropriateness categories. Blacks were significantly less likely to undergo percutaneous transluminal coronary angioplasty (odds ratio, 0.30; 95% confidence interval, 0.14-0.63 [P<.01]) when the indication was rated "equivocal." Similarly, blacks were less likely to undergo coronary artery bypass grafting (odds ratio, 0.44; 95% confidence interval, 0.23-0.86 [P<.01]) when only coronary artery bypass grafting was indicated as "appropriate and necessary." Differences in comorbidity or use of cigarettes or alcohol did not explain these variations. Using administrative data from the Veterans Health Administration, we found no differences in 1-year (5.2% vs 7.4%) and 5-year (23.3% vs 26.2%) mortality for blacks vs whites. CONCLUSION: Among patients with acute myocardial infarction or unstable angina, variation in clinical factors using RAND appropriateness criteria for procedures explained some, but not all, racial differences in coronary revascularization use. PMID- 10809038 TI - Aortic valve replacement in patients with aortic stenosis and severe left ventricular dysfunction. AB - BACKGROUND: The outcome of aortic valve replacement for severe aortic stenosis is worse in patients with impaired left ventricular function. Such dysfunction in aortic stenosis may be reversible if caused by afterload mismatch, but not if it is caused by superimposed myocardial infarction. METHODS: From our echocardiography database, 55 patients with severe aortic stenosis (valve area < or =0.75 cm2) and ejection fractions of 30% or lower who subsequently underwent aortic valve replacement were included. The operative mortality and clinical follow-up were detailed. RESULTS: There were 10 perioperative deaths (operative mortality, 18%). Twenty (36%) of the 55 patients had a prior myocardial infarction. In the 35 patients without prior myocardial infarction, there was only 1 death (3%). In contrast, 9 of 20 patients with prior myocardial infarction died (mortality rate, 45%; P< or =.001). The factors significantly associated with perioperative death on univariate analysis (functional class, mean aortic gradient, and prior myocardial infarction) were entered into a model for stepwise logistic regression. This multivariate analysis showed that only prior myocardial infarction was independently associated with perioperative death (odds ratio, 14.9; 95% confidence interval, 2.4-92.1; P = .004). CONCLUSIONS: The risk of aortic valve replacement in patients with severe aortic stenosis and severely reduced left ventricular systolic function is extremely high if the patients have had a prior myocardial infarction. This information should be factored into the risk-benefit analysis that is done preoperatively for these patients, and it may preclude operation for some. PMID- 10809039 TI - Premature cardiovascular disease is common in relatives of patients with premature peripheral atherosclerosis. AB - BACKGROUND: Numerous clinical conditions have been proposed to explain the premature onset of symptomatic peripheral vascular disease (PVD) in young adults, but the role of genetic factors has not been defined. This study was performed to determine the prevalence of cardiovascular disease among family members of patients with premature PVD. METHODS: The prevalence of early cardiovascular events occurring in first-degree relatives of 90 subjects with premature PVD (onset < or =49 years) was determined. The prevalence of occult atherosclerosis was determined by duplex ultrasonography in a cohort of 20 asymptomatic siblings. Reference groups included first-degree relatives of 80 subjects with premature coronary artery disease (CAD) and first-degree relatives of 48 healthy subjects. RESULTS: Cardiovascular events occurred at age 55 years or younger in 28% of the parents of PVD subjects, in 23% of parents of CAD subjects, and in 7% of the parents of healthy controls (P<.001). Cardiovascular events occurred in 24% of siblings of PVD subjects, in 14% of siblings of CAD subjects, and in 7% of siblings of healthy controls (P<.001). Duplex ultrasonography detected early plaques in the lower extremity circulation of 10 (50%) of the asymptomatic siblings of PVD subjects. CONCLUSIONS: Early, symptomatic cardiovascular disease is more common in first-degree relatives of individuals with premature PVD than in relatives of healthy individuals or of probands with premature CAD. Occult vascular disease in the lower extremity is prevalent among asymptomatic siblings of probands with premature PVD. These observations indicate that susceptibility to premature PVD has a familial basis. PMID- 10809040 TI - Helicobacter pylori--is it a novel causative agent in Vitamin B12 deficiency? AB - BACKGROUND: Evidence for vitamin B12 deficiency usually involves combinations of low serum vitamin B12 levels, clinical and metabolic abnormalities, and therapeutic response. Identification of the underlying cause is important in the diagnosis of vitamin B12 deficiency that is usually attributed to malabsorption. Helicobacter pylori is one of the most common causes of peptic ulcer disease worldwide and a major cause of chronic superficial gastritis leading to atrophy of gastric glands. It is suggested that there may be a casual relationship between H. pylori and food-cobalamin malabsorption. OBJECTIVES: To evaluate the H. pylori incidence in patients with vitamin B12 deficiency prospectively and to assess whether treatment for H pylori infection could correct this deficiency over time. PATIENTS AND METHODS: We performed a prospective cohort study involving 138 patients who had anemia and vitamin B12 deficiency. An upper gastrointestinal endoscopy was performed to assess the severity of atrophic gastritis and biopsy specimens for Campylobacter-like organisms tests and histological examination for H pylori were obtained at the time of diagnosis. The diagnosis of H. pylori prompted a combination treatment. RESULTS: Helicobacter pylori was detected in 77 (56%) of 138 patients with vitamin B12 deficiency and eradication of H pylori infection successfully improved anemia and serum vitamin B12 levels in 31 (40 %) of 77 infected patients. CONCLUSIONS: Helicobacter pylori seems to be a causative agent in the development of adult vitamin B12 deficiency. Eradication of H. pylori infection alone may correct vitamin B12 levels and improve anemia in this subgroup of patients. PMID- 10809042 TI - Impact of evidence-based "clinical judgment" on the number of American adults requiring lipid-lowering therapy based on updated NHANES III data. National Health and Nutrition Examination Survey. AB - BACKGROUND: When the National Cholesterol Education Program Adult Treatment Panel II (ATP II) guidelines were published, National Health and Nutrition Examination Survey III data for 1988 to 1991 were used to estimate the number of Americans requiring lipid-lowering therapy based on ATP II cut points. However, the guidelines recommend using clinical judgment to determine whether to initiate drug therapy in individuals whose low-density lipoprotein cholesterol levels remain above treatment goals with diet therapy but below the initiation level for drug therapy. METHODS: We analyzed updated (1988-1994) National Health and Nutrition Examination Survey III data, based on a sample of 6796 adults aged 20 years and older, to estimate the numbers of American adults with an elevated low density lipoprotein cholesterol level and requiring drug therapy using cut points vs clinical judgment as specified in ATP II guidelines. RESULTS: Assuming a 10% low-density lipoprotein cholesterol reduction with diet, an estimated 10.4 million American adults require drug therapy based on ATP II cut points. If we include individuals for whom the guidelines recommend clinical judgment, the estimate increases to 28.4 million. The largest increase occurs in individuals without known coronary heart disease but with 2 or more risk factors: from 5.5 to 17.5 million. These high-risk individuals have low-density lipoprotein cholesterol concentrations similar to those in patients with coronary heart disease. CONCLUSIONS: Since the ATP II guidelines were published, clinical judgment has been informed by abundant clinical trial evidence establishing the safety and benefit of lipid-lowering therapy. The large number of individuals at high risk for coronary heart disease emphasizes the need for cost-effective therapy to extend treatment to the greatest number of individuals who may benefit. PMID- 10809041 TI - Depression and 1-year prognosis in unstable angina. AB - BACKGROUND: Depression is common after acute myocardial infarction and is associated with an increased risk of mortality for at least 18 months. The prevalence and prognostic impact of depression in patients with unstable angina, who account for a substantial portion of acute coronary syndrome admissions, have not been examined. METHODS: Interviews were carried out in hospital with 430 patients with unstable angina who did not require coronary artery bypass surgery before hospital discharge. Depression was assessed using the 21-item self-report Beck Depression Inventory and was defined as a score of 10 or higher. The primary outcome was 1-year cardiac death or nonfatal myocardial infarction. RESULTS: The Beck Depression Inventory identified depression in 41.4% of patients. Depressed patients were more likely to experience cardiac death or nonfatal myocardial infarction than other patients (odds ratio, 4.68; 95% confidence interval, 1.94 11.27; P<.001). The impact of depression remained after controlling for other significant prognostic factors, including baseline electrocardiographic evidence of ischemia, left ventricular ejection fraction, and the number of diseased coronary vessels (adjusted odds ratio, 6.73; 95% confidence interval, 2.43-18.64; P<.001). CONCLUSIONS: Depression is common following an episode of unstable angina and is associated with an increased risk of major cardiac events during the following year. PMID- 10809043 TI - Hemolytic uremic syndrome associated with clopidogrel: a case report. AB - Clopidogrel and ticlopidine are antiplatelet agents used in the treatment of patients with cerebrovascular and peripheral vascular disease and to reduce the risk for thrombosis in patients undergoing coronary artery stenting. Ticlopidine has been reported to have major hematologic adverse effects, including neutropenia and thrombotic thrombocytopenic purpura or hemolytic uremic syndrome (HUS). Clopidogrel, an analogue of ticlopidine, was developed because it did not show bone marrow toxic effects in either tissue culture or animal models. In human studies, to date, clopidogrel has been associated with a low incidence of severe neutropenia and no reported cases of thrombotic thrombocytopenic purpura or HUS. For these reasons, clopidogrel has been increasingly used in place of ticlopidine after coronary artery stenting. We report a case of clopidogrel associated HUS. This observation implicates clopidogrel as a possible causative agent in HUS. PMID- 10809044 TI - Hallucinations in an elderly patient taking recommended doses of cyclobenzaprine. PMID- 10809045 TI - Somnum fuge meridianum. PMID- 10809046 TI - Homeopathy: alternative, but not scientific. PMID- 10809047 TI - Is homeopathy incompatible with evidence-based medicine? PMID- 10809048 TI - Homeopathy wrongly revisited? PMID- 10809049 TI - Homeopathy: one more revision. PMID- 10809050 TI - When should lipids be measured in myocardial infarction? PMID- 10809051 TI - Does extreme dipping of nocturnal blood pressure in elderly hypertensive patients confer high risk of developing ischemic target organ damage from antihypertensive therapy? PMID- 10809052 TI - The target antigens of naturally occurring human anti-beta-galactose IgG are cryptic on porcine aortic endothelial cells. AB - The identification of the xeno-antigens/xeno-antibodies combinations involved in pig-to-human xenograft rejection is an essential step for understanding this process and for the development of procedures to prevent it. Although it is widely accepted that the terminal disaccharide Galalpha1,3Gal-R is by far the major epitope recognized by human natural antibodies reactive with pig tissues, there is also evidence that other carbohydrate epitopes might be important in xenograft rejection. In an attempt to further improve our knowledge of the repertoire of human natural antibodies with anti-pig specificity we sought to determine whether naturally occurring human anti-beta-galactose IgG could interact with porcine aortic endothelial cells (PAEC). Histochemical analysis of porcine aorta sections revealed that the carbohydrate structures recognized by the anti-beta-galactose IgG are present on endothelial cells but in a cryptic form that can be unmasked by sialidase treatment. These structures were also found to be cryptic in cultured PAEC. In addition we demonstrated that PAEC may adsorb fetal calf serum (FCS) glycoproteins when cultured in FCS-supplemented medium, a process susceptible to generating artifactual observations in carbohydrate antigens analysis. In conclusion, despite their abundance, human anti-beta-galactose IgG do not represent a primary concern in pig-to-human xenotransplantation as the carbohydrate structures to which they bind are normally masked by sialic acid residues on porcine endothelial cells. However, whether these cryptic epitopes might be exposed on endothelial cells from genetically engineered animals should be further investigated because, if so, additional approaches will be needed to suppress their interaction with human anti-beta-galactose IgG. PMID- 10809053 TI - Removal of xenoreactive antibodies by protein-A immunoadsorption: experience in 22 patients. AB - The presence of naturally occurring anti-Galalpha1-3Gal (antialphaGal) Ab in human serum is believed to be a major factor in the pathogenesis of hyperacute rejection of discordant organ xenografts such as the pig-to-human combination. Galalpha1-3Gal epitopes are expressed on pig tissues and the binding of anti Galalpha1-3Gal leads to endothelial cell activation and complement-mediated hyperacute graft rejection. Several strategies have been suggested in donor animals or in the xenograft recipient to overcome the anti-alphaGal barrier. Protein-A immunoadsorption (PAIA) was developed for the in vivo removal of circulating Ab and it has been shown to be effective in cases where pathogenic auto or alloAb are present. The aim of our study was to analyze the effect of PAIA on total and xenoreactive serum anti-alphaGal immunoglobulin levels in a group of patients treated with this technique for different diseases. After three consecutive sessions of PAIA, total and xenoreactive IgG and IgM immunoglobulin levels were decreased by more than 50% of pre-treatment levels. So we conclude that PAIA is an effective method to significantly reduce circulating Ab, including xenogeneic IgM and IgG Ab. This mode of therapy might be considered as a tool to overcome hyperacute xenograft rejection. PAIA combined with other therapeutic approaches may well protect the xenograft. PMID- 10809054 TI - Human serum induces apoptosis of xenogenic cardiomyocytes in vivo and in vitro. AB - Discordant xenotransplantation is complicated by delayed xenograft rejection (DXR). Previous studies have demonstrated that anti-apoptotic genes are protective against DXR. This study examines the hypothesis that apoptosis plays a role in human anti-xenograft responses. C57BL/6 mice and NOD SCID mice were given a single intravenous injection of either a lethal dose (LD, survival < 30 min) or a sublethal dose (SLD) of human serum, and isolated pig and mouse rod-shaped cardiomyocytes were exposed to human serum in vitro. In situ detection of apoptotic cells in mouse hearts was assessed using a terminal deoxynucleotidyl transferase-mediated dUTP nicked-end labeling assay. Mice transfused with human serum had approximately a 10-fold increased percentage of apoptotic cells after SLD 18 h post-injection compared with animals given saline, and a fourfold increase over LD. Administration of cobra venom factor (CVF) decomplemented SLD 18 h did not significantly (P > 0.05) alter the percentage apoptosis. The addition of 20 mM Gal-alpha-1,3-Gal to SLD 18 h significantly (P < 0.05) reduced percentage apoptosis to levels comparable to saline treated control animals. In vitro using mouse and pig cardiomyocytes demonstrated parallel results as in vivo experiments. Human serum induces apoptosis of cardiomyocytes in immunocompetent and immunoincompetent mice in vivo, as well as mouse and pig cardiomyocytes in vitro. Further, this apoptotic response can be inhibited by the addition of Gal alpha-1,3-Gal without affecting the capacity of the serum to cause HAR. These results demonstrate that a putative human serum factor induces a delayed apoptotic injury of xenograft tissues, and supports the hypothesis that apoptosis may be an important mediator of DXR. PMID- 10809055 TI - Venular thrombosis is the key event in the pathogenesis of antibody-mediated cardiac rejection. AB - A review of the histopathologic features of serial biopsies and excised grafts of 117 experimental and clinical cardiac allografts and xenografts revealed a common sequence in the development of histopathologic changes in grafts showing antibody mediated (hyperacute and acute vascular) rejection. Based on these observations, we propose the new concept that thrombosis of cardiac veins and venules is the initial key event in antibody-mediated rejection. This is followed by the development of congestion in the subtended venules and capillaries accompanied by interfascicular and, later, intermyocyte edema. Subsequently, focal or diffuse interstitial hemorrhage affecting the subendocardium, extending sometimes to involve the inner half of the ventricular myocardium, is observed. Antibody mediated rejection therefore appears to be analogous to incomplete venous infarction of the heart. The observed histopathology (in which venular thrombosis plays a key role) favors a thrombogenic basis for the classical features of antibody-mediated rejection, namely edema, vascular thrombi and interstitial hemorrhage. A key role for venular thrombosis would explain the non-uniform distribution of the changes and may suggest new ways of preventing antibody mediated xenograft rejection. PMID- 10809056 TI - Naturally acquired anti-alpha Gal antibodies in a murine allograft model similar to delayed xenograft rejection. AB - Antibodies directed against galactose-alpha1,3-galactose (alphaGal) are believed to play an important role in the pathogenesis of delayed xenograft rejection (DXR). This study was designed to determine whether alpha1,3 galactosyltransferase-deficient (Gal KO) mice can naturally acquire a sufficient anti-alphaGal titre to cause the delayed type rejection of alphaGal-expressing hearts. Gal KO mice of various ages were assessed for anti-alphaGal antibody levels. alphaGal-expressing hearts were transplanted heterotopically into these mice and monitored daily. Rejecting and surviving hearts were evaluated histologically. In Gal KO mice greater than 6-month-old, 64% had an anti-alphaGal antibody titre above the background level. When wild-type alphaGal-expressing hearts were transplanted into this group, 45% of grafts rejected within 5 to 13 days. Histological examination of the rejected hearts displayed marked tissue damage and an inflammatory infiltrate of predominantly macrophage/monocytes. Surviving grafts showed preserved morphology. Like humans, Gal KO mice naturally develop anti-alphaGal antibodies with age. The titre in these mice was sufficient to cause a "delayed-type" rejection of a significant proportion of alphaGal expressing cardiac grafts. This model thus provides an opportunity to investigate the role of naturally acquired anti-alphaGal antibodies in the pathogenesis of DXR. PMID- 10809057 TI - Tolerance of T-independent xeno-antibody responses in the hamster-to-rat xenotransplantation model is species-restricted but not tissue-specific. AB - Control of early acute xenograft rejection xenoreactions in the hamster-to-Lewis rat xenotransplantation model with cyclosporine (CsA) and leflunomide subdues early T-independent xenoreactivity and uncovers a late immune response that can be controlled by CsA alone. We had attributed this acquired responsiveness to CsA to the induction of tolerance of T-independent xeno-antibody responses in the recipient and recently reported that this tolerance is species-specific. Here we have further characterized the specificity and nature of this tolerant state. Lewis rats transplanted with either hearts, skin, kidney or spleen/pancreas from Golden Syrian hamsters were treated with leflunomide (5 mg/kg/day by gavage) for 14-21 days and CsA (20 mg/kg/day by gavage) continuously from the day of transplant. Some Lewis rats received a second graft of hearts or skin from Golden Syrian hamsters (day 21-30 after first transplant), and a third heart graft from Balb/c mice (day 60 after the first transplant). Serum was harvested and the titers of xenoreactive antibodies were quantified by flow cytometry. All grafts were harvested at the end of each experiment and examined by histological and immunohistochemical methods. The combination of CsA and leflunomide was able to completely inhibit the rejection of kidney, spleen and pancreas xenografts in this hamster-to-rat xenotransplantation model. In addition, only a transient treatment with leflunomide was necessary, and long-term graft survival could subsequently be maintained by CsA alone. Histological examination of these grafts at > 80 days post-transplantation indicated minimal signs of rejection. These immediately vascularized organs induced T-independent B-cell tolerance, so that second grafts of hamster hearts and skin could be maintained with CsA alone. Under the same immunosuppressive regimen, only four out of nine Lewis rats exhibited long-term hamster skin survival, probably reflecting the increased immunogenicity of the skin compared with other vascularized grafts. Nonetheless, all rats that did not reject the hamster skin graft also did not reject the hamster heart while on CsA alone. Finally, we demonstrate that the tolerant state could be maintained for up to 30 days in the absence of xenograft. The vigorous T independent antibody response that mediates acute xenograft rejection in the hamster-to-rat model can be tolerized by the immunosuppressive regimen of CsA and leflunomide. The lack of organ specificity for the induction of this tolerance suggests that the xenoantigens inducing tolerance may be common endothelial cell antigens. Finally, the presence of the xenograft has been previously shown to be critical for the induction of T-independent B-cell tolerance, however, the tolerant state is relatively stable and persists after the removal of the xenograft. PMID- 10809058 TI - Development and analysis of transgenic mice expressing porcine hematopoietic cytokines: a model for achieving durable porcine hematopoietic chimerism across an extensive xenogeneic barrier. AB - The capacity of mixed hematopoietic chimerism to induce tolerance has not been demonstrated in discordant xenogeneic species combinations because of the difficulty in achieving lasting hematopoietic engraftment. In an effort to create a model of long-lasting disparate xenogeneic hematopoietic chimerism, we have developed transgenic (Tg) mice carrying porcine cytokines. Three lines of Tg mice were generated: one carrying porcine IL-3 and GM-CSF genes only (termed IL/GM) and the remaining two lines carrying in addition, the soluble SCF gene (termed IL/GM/sS) or membrane-bound SCF gene (termed IL/GM/mS). Sera from mice with IL/GM and IL/GM/sS transgenes markedly stimulated the proliferation of swine marrow cells in vitro. However, proliferation of swine marrow cells was not induced in cultures containing IL/GM/mS sera. Consistent with these observations, ELISA assays revealed detectable levels of porcine cytokines in the sera of IL/ GM and IL/GM/sS, but not in sera of IL/GM/mS Tg mice. Marrow stromal cells prepared from all three kinds of Tg mice, but not those from non-Tg littermates, were capable of supporting the growth of porcine hematopoietic cells in vitro. Immunodeficient Tg mice were generated by crossing Tg founders with C.B-17 SCID mice for five generations. All Tg immunodeficient mice showed improved porcine hematopoietic engraftment compared with non-Tg controls. These Tg mice provide a useful model system for studying porcine hematopoietic stem cells, and for evaluating the feasibility of donor-specific tolerance induction by mixed chimerism across highly disparate xenogeneic barriers. PMID- 10809059 TI - Synthesis and ESR studies of redox reactivity of bis (3,5-di-tert-butyl-1,2 benzoquinone-2-monooximato)Cu(II). AB - Complexing of 3,5-di-tert-butyl-1,2-benzoquinone-2-monooxime with Cu(II) in air and under N2 gave Cu(qo)2 and Cu(qo)2 x H2O (where qo is 3,5-di-tert-butyl-1,2 benzoquinone-2-monooximato-anion) complexes, respectively. The ESR spectroscopy showed that the reduction of these complexes with P(PhX)3 (X = H, m-Cl, m-CH3, p Et2N-) and 1,4-bis(diphenyldiphosphino) butane (dppb) proceeds via the radical formation (phenoxazine, amino phenoxy and nitrene type radical intermediates) and pathways of reduction depend on the structure of these complexes. The reaction of Cu(qo), with dppb and P(PhX)3 phosphines gave essentially identical ESR spectra. At the same time, reduction of Cu(qo)2 x H2O with PPh3 result in entirely different unstable radical spectrum (g = 2.0046) which is further converted to another relatively stable Cu-containing radical signal (g = 2.0052). The unstable radical species attributed to nitrene type radicals. The initial complexes and all radical products were characterized by their ESR and optical spectra. PMID- 10809060 TI - Reassignment of fundamental vibrational modes of cyclic S4N3 cation. AB - MP2/6 31G* calculations were carried out to investigate the vibrational spectrum of cyclic S4N3+. The results indicate that previous assignments of several fundamental vibrational modes are in error. On the basis of the calculated results, reassignments of these modes are proposed. PMID- 10809061 TI - Raman excitation profiles and excited state molecular configurations of three isomeric phenyl pyridines. AB - Contributions of different electronic states to Raman scattering have been studied by critical analyses of Raman excitation profiles (REPs) of several normal modes of vibration of three isomeric phenyl pyridines. In this context, possible structures and other interesting properties of the three molecules in the excited electronic states have been discussed. Normal mode characteristics are also described. Most likely a singlet state, lying in the vacuum ultraviolet region with respect to the ground state, is found to be playing a very significant role in the scattering phenomena. PMID- 10809062 TI - Vibrational spectra of four alkaline earth cyclo-hexaphosphates. AB - FT-IR and FT-Raman spectra of four alkaline earth (Mg, Ca, Sr and Ba) cyclo hexaphosphates have been recorded and analysed. FT-Raman spectra of the deuterated analogues of these compounds are used to clear the ambiguity regarding the OH vibrations in the low frequency region. The spectra reveal that the P6O18(6) anion ring in all compounds have independent PO4 tetrahedra present in it. The P-O(P)and P-O(M')(M'-Mg, Ca, Sr and Ba) bonds become stronger as the cation size decreases. The P6O18(6-) anion ring is distorted in the Mg and Ca compounds. In the Sr and Ba compounds, free and hydrogen bonded water molecules of varying strength are present, whereas in Mg and Ca compounds no free water molecules exist. The POP bridge angle of the Mg, Ca and Sr compounds are estimated using the correlations between the POP bridge stretching frequencies and the bridge angle value. PMID- 10809063 TI - Infrared and polarized Raman spectra of (CH3)2NH2Al(SO4)2 x 6H2O. AB - FTIR and single crystal Raman spectra of (CH3)2NH2Al(SO4)2 x 6H2O have been recorded at 300 and 90 K and analysed. The shifting of nu1 mode to higher wavenumber and its appearance in Bg species contributing to the alpha(xz) and alpha(yz) polarizability tensor components indicate the distortion of SO4 tetrahedra. The presence of nu1 and nu2 modes in the IR spectrum and the lifting of degeneracies of nu2, nu3, and nu4 modes are attributed to the lowering of the symmetry of the SO4(2-) ion. Coincidence of the IR and Raman bands for different modes suggest that DMA+ ion is orientationally disordered. One of the H atoms of the NH2 group of the DMA+ ion forms moderate hydrogen bonds with the SO4(2-) anion. Al(H2O)6(3+) ion is also distorted in the crystal. The shifting of the stretching modes to lower wavenumbers and the bending mode to higher wavenumber suggest that H2O molecules form strong hydrogen bonds with SO4(2-) anion. The intensity enhancement and the narrowing of nu1SO4, deltaC2N and Al(H2O)6(3+) modes at 90 K confirm the settling down of the protons in the hydrogen bonds formed with H2O molecules and NH2 groups. This may be one of the reasons for the phase transition observed in the crystal. PMID- 10809064 TI - Extraction of diffuse reflection spectrum on reflectance spectroscopy for solid or powder surfaces. AB - Reflectance spectroscopy usually produces distorted spectra because it is arbitrarily composed of two different types of reflectance, specular reflectance and diffuse reflectance. That is why this has been an annoying problem for reflectance spectroscopists. In this report, we studied the extraction of only the diffuse reflectance spectra from several reflectance spectra using redundant multiple spectral information, non-negative transformation of the recorded spectra and information processing as the multi-curve resolution. As a result, we found that we could estimate just the diffuse reflectance spectral profile. PMID- 10809065 TI - Laser flash photolysis study of triphenylimidazole. AB - Laser flash photolysis of the photocyclization of triphenylimidazole (TPI) in ethyl alcohol at 308 nm. indicates that the dihydrophenanthroimidazole (DHPI) intermediate is produced rapidly, has a lifetime of 0.25 ms, and returns predominantly back to triphenylimidazole. Analysis of the decay channels for this intermediate indicates two rate constants: (1) k1 = 3.3 x 10(3) s(-1), associated with reversion back to triphenylimidazole and (2) k2 = 0.67 x 10(2) s(-1), which is associated with the conversion of the dihydrophenanthroimidazole to the photoproduct, 2-phenyl-9,10-phenanthroimidazole. The photoproduct is readily observed as an increasing component in the biexponential fluorescence decay data. Fluorescence lifetimes for triphenylimidazole and 2-phenyl-9,10 phenanthroimidazole (PPI) in ethyl alcohol were determined to be 1.76 and 8.21 ns, respectively, with no additional components in the fluorescence decay as the photochemistry proceeds. An additional transient absorption observed in the 450 nm. region, with a lifetime of 0.7 micros, decaying faster than the dihydrophenanthroimidazole intermediate, is assigned to the triplet state of triphenylimidazole. PMID- 10809066 TI - Hyperfine interaction in K2Ba[Fe(NO2)6]. AB - Magnetic hyperfine splitting observed in the low temperature Mossbauer spectrum of potassium barium hexanitro ferrate(II), in the absence of any external field, is attributed to the 5T2g state of the central metal atom further split into a ground 5Eg state and a first excited 5B2g state under a distorted octahedral symmetry in contrast to the earlier prediction of 1A1g ground state on the basis of room temperature Mossbauer spectral and other properties. The central iron atom is coordinated to six nitrito groups (NO2-), having an oxidation state of +2. The temperature dependence of Mossbauer spectra is explained on the basis of electronic relaxation among the spin-orbit coupled levels of the 5Eg ground state. Various kinds of electronic relaxation mechanisms have been compared to explain the proposed mechanism. The observed temperature dependent spectra with varying internal magnetic field and line width can be explained by simple spin lattice relaxation. PMID- 10809067 TI - Effect of micelles on the spectral characteristics of the prototropic species of 2-(2'-aminophenyl)benzimidazole. AB - The absorption and fluorescence spectra of 2-(2'-aminophenyl)benzimidazole (2 APBI) have been studied in anionic (sodium dodecylsulphate, SDS), cationic (cetyltrimethylammonium bromide, CTAB) and non-ionic micelles (Tween-80, TritonX 100) at different acid-base concentrations. Spectral characteristics of 2-APBI at various acid concentrations have established only one kind of monocation (MC) in Tween-80 and TritonX-100 (TX-100), whereas two kinds of MCs are present in SDS. Above study has further shown that there is a strong hydrogen bonding interaction between the polar polyoxyethylene groups of non-ionic micelles and the MCs of 2 APBI. This interaction is responsible for the stabilization of the MC III to MC III' which is more planar than either MC II or MC III, which is otherwise less stable in water. This is substantiated by the lifetime data, fluorescence excitation spectra and the pKa values of the monocation-neutral (MC-N) equilibrium. PMID- 10809068 TI - Transverse and longitudinal crystal modes associated with OH stretching vibrations in single crystals of kaolinite and dickite. AB - Raman spectra of randomly-oriented kaolinite, dickite and nacrite show, for coarsely crystalline material, an extra band in the OH stretching region which is absent from the IR spectra of clay-size samples. Oriented single-crystal Raman spectra of these minerals provide confirmation for the assignment of the extra bands to transverse optical modes involving in-phase coupled vibrations of the layer-surface hydroxyl groups. The corresponding IR bands have transition moments nearly perpendicular to the layer surface, and appear at the higher frequencies of the longitudinal optical modes of macroscopic crystals. PMID- 10809069 TI - Raman spectroscopy of nacrite single crystals at 298 and 77 K. AB - A Raman microscope in conjunction with a thermal stage has been used to determine the Raman spectra of single crystals of nacrite at 298 and 77 K. The spectra obtained are a function of the physics of the spectrometer and were orientation dependent. Bands are observed at 3710, 3646, 3630 and 3623 cm(-1). Upon obtaining the Raman spectra at liquid nitrogen temperature, the band at 3648 cm(-1) was not observed but an additional band at 3603 cm(-1) appeared. This latter band may be attributed to the hydroxyl stretching of non-hydrogen bonded interlayer hydroxyls in the nacrite. The bands attributed to both the inner and inner surface hydroxyls moved to lower frequencies upon cooling to liquid nitrogen temperatures. Low frequency bands also showed orientation dependence. PMID- 10809070 TI - [UV absorption spectra of 1,1'-binaphtyle and 2,2'-binaphtyle molecules and their self-associations from the atom monopole-dipole interaction (AMDI) model]. AB - The Atom Monopole-Dipole interaction (AMDI) model is used for the first time in calculation of UV absorption spectra of 1,1'-binaphtyl and 2,2'-binaphtyl and their self-associations for the case of frequency-dependant atom polarisability with Lorentzian band shape. UV absorption spectra are obtained from a set of electronic normal modes. The variation of spectra with the conformation of these molecules concord with the results of the medium effect on the corresponding electronic transitions. The calculated auto-association spectra are qualitatively in good agreement with polycyclic aromatic hydrocarbon aggregations behaviour. PMID- 10809071 TI - Vibrational anharmonicity and vibrational Stark effect of sulfate ions trapped in potassium, rubidium and cesium chromate. AB - Room and low temperature (approximately 100 K) FT-IR and Raman spectra of the sulfate doped K2CrO4, Rb2CrO4 and Cs2CrO4 were recorded. The positions of the nu1, nu3, and nu4 fundamental mode components of the dopant anions were measured. Nine (out of possible ten) second-order stretch-stretch vibrational transitions of the dopant anions were detected. On the basis of these data, the anharmonicity constants and the corresponding harmonic eigenvalues were calculated for several vibrational transitions using second-order perturbation theory expressions. The anharmonicity of the studied second-order transitions of the type nu1 + nu3i increases in the order (SO4/K2CrO4) < (SO4/Rb2CrO4) < (SO4/Cs2CrO4), while for those of the type nu3j + nu3i it basically follows the trend: (SO4/K2CrO4) > (SO4/Rb2CrO4) < (SO4/Cs2CrO4). The measured relative Stark splittings of the nu3 and nu4 mode components of the dopant SO4(2-) anions, as well as the average X13i/3i3j values decrease in the order (SO4/K2CrO4) > (SO4/Rb2CrO4) > (SO4/Cs2CrO4). In all cases, the splitting is larger for nu3 than for nu4 modes, indicating a smaller angular than the bond length distortion. The theory of vibrational Stark effect suggests that the observed frequency shifts of the nu1, nu3 and nu4 mode components may be attributed to the increase of the field strength at the doped anion site going from K2CrO4 to Cs2CrO4. The Stark splitting of these modes, on the other hand, implies that the internal crystalline field vector is almost parallel to the (hypothetical) C2 axis of the slightly distorted dopant tetrahedral anions. PMID- 10809072 TI - Surface-enhanced Raman scattering (SERS) studies on 1,1'-bi-2-naphthol. AB - The surface-enhanced Raman (SER) spectrum of 1,1'-bi-2-naphthol was obtained using silver colloids. Comparison of the SER spectrum with the spectrum in solution and that of the solid are made. 1,1'-Bi-2-naphthol is thought to adsorb in a tilted position relative to the silver surface. PMID- 10809073 TI - Electronic absorption spectra of benzoquinone and its hydroxy substituents and effect of solvents on their spectra. AB - The electronic absorption spectra of 1,4-benzoquinone (BQ) and its 2,5-dihydroxy and tetrahydroxy derivatives have been studied in detail. The interpretation of the electronic bands is made on the basis of PPP and CNDO calculations. It is found that the pi --> pi* transitions are well predicted by the PPP method. The predictions of the CNDO method are however superior both in their accuracy as well as ability to predict the n --> pi* transitions. The effect of solvents on the electronic absorption bands have also been investigated in detail. Linear correlations are found between the solvent's dielectric constant and wavelength of the absorption bands. The solvent shifts are explained on the basis of the polarities of the solute and solvent molecules as well as due to hydrogen bonding. PMID- 10809074 TI - Effect of solvents on the photophysical properties of substituted imidazonaphthyridine derivatives. AB - The absorption and fluorescence characteristics of three substituted imidazonaphthyridine derivatives are studied in a series of organic solvents. The variation of Stokes shift with the polarity parameter of the solvent is studied and the excited state dipole moment of these derivatives is higher than the ground state dipole moment. The fluorescence lifetime profile shows single exponential decay in all the solvents. The fluorescence quantum yield, radiative and non-radiative rate constants are also calculated and these parameters show much variation in the change in substitution of these derivatives. PMID- 10809075 TI - High resolution FTIR spectrum of the nu1 band of DCOOD. AB - Accurate spectral information on formic acid has wide application to radioastronomy since it was the first organic acid found in interstellar space. In this work, the infrared absorption spectrum of the nu1 band of deuterated formic acid (DCOOD) has been measured on a Bomem DA3.002 Fourier transform spectrometer in the wavenumber region 2560-2690 cm(-1) with a resolution of 0.004 cm(-1). A total of 292 infrared transitions have been assigned in this hybrid type A and B band centred at 2631.8736 +/- 0.0004 cm(-1). The assigned transitions have been fitted to give a set of eight rovibrational constants for the nu1 = 1 state with a standard deviation of 0.00078 cm(-1). PMID- 10809076 TI - Photophysics of Nile red in solution: steady state spectroscopy. AB - Spectroscopic properties of Nile red (NR) in organic solvents, binary solvent mixtures have been studied. Remarkable shifts in the emission band positions have been observed as a function of the polarity of the medium. In solvent mixtures, these shifts can be explained by the process of specific solvation known as dielectric enrichment. The displacement of the fluorescence band was also measured as a function of temperature to obtain the thermochromic shifts (15 cm( 1) K(-1) in methyltetrahydrofuran and 13.8 cm(-1) K(-1) in butanol). Excited state dipole moments were calculated from these shifts. PMID- 10809077 TI - Application of aluminium(III) complex with salicylidene-o-aminophenol to the fluorometric determination of nucleic acids. AB - In buffer medium of hexamethylene tetraamine-HCl at pH 5.9 the aluminium(III) complex with salicylidene-o-aminophenol (SAP) has a fluorescence peak at 508 nm with excitation at 410 nm. When nucleic acid coexists, it reacts with the complex within 8 min at room temperature to produce a non-fluorescent product, resulting in the decrease of fluorescence intensity of the aluminium complex. On basis of this, a new fluorometric method for nucleic acids determination is proposed. The calibration graphs for calf thymus DNA, fish sperm DNA and yeast RNA are linear up to 5.0, 4.0 and 3.0 microg ml(-1), respectively, and corresponding detection limits are 49, 52 and 62 ng ml(-1). The synthetic samples are analyzed with relative standard deviation of five measurements of 3.9-6.0%. DNA in an extraction product from human blood is determined using the calibration graph for calf thymus DNA, and the result is very close to that by the ethidium bromide assay. Compared with some established fluorometric methods, this procedure is sensitive, selective, reliable, reproducible and practical. The association constant of calf thymus DNA with the complex is estimated by two graphic methods. It is suggested that the binding reaction between nucleic acids with the complex proceeds in an intercalation way. PMID- 10809078 TI - Determination of proteins by fluorescence quenching of Magdala Red. AB - Magdala Red (MR) binding to protein causes a decrease in the fluorescence intensity of MR at 556 nm. Based on this, a new quantitative determination method for proteins is developed. The linear range of this assay is 0.1-4.0 microg ml( 1) of Bovine Serum albumin (BSA). The measurements can be made easily on a common fluorimeter. The reaction between MR and proteins is completed in 1 min, and the fluorescence intensity is stable for at least 2 h. There is little or no interference from amino acids and most metal ions. The proposed method has been applied to the determination of protein in milk powder and soybean milk powder and the results are in agreement with the results by the other methods. PMID- 10809079 TI - Carbazole as an excited state proton transfer fluorescent probe for lipid bilayers in alkaline medium. AB - In our effort to look for novel excited state proton transfer (ESPT) fluorescent probes in alkaline pH range, we have examined carbazole as a possible candidate because of its high extinction coefficient, high quantum yield and a larger difference in ionization constant between the ground and excited state (pKa - pKa*). The photodissociation of carbazole was studied in liposome membrane by steady state fluorescence measurements at alkaline pH ranges. The neutral form and the anionic form of carbazole emit at 362 and 417 nm, respectively. This large shift in emission makes it convenient to monitor the physical properties of liposomes. The neutral form fluorescence intensity of carbazole is sensitive to phase changes in the membrane and also shows a maximum at phase transition temperature. This variation of intensity can be explained in terms of redistribution of probe between the surface and interior of the liposomes. Cholesterol induced phase changes of liposomes were also sensed by the ESPT of carbazole. PMID- 10809080 TI - Structure and function of the Groucho gene family and encoded transcriptional corepressor proteins from human, mouse, rat, Xenopus, Drosophila and nematode. AB - A gene family of the Groucho, TLE, ESG and AES proteins has been characterized from Drosophila, nematode, Xenopus, mouse, rat and human, and their structural relationships have been analyzed. The genomic organization of nematode ESG, human and mouse AES genes has been determined, and the expression of ESG and AES genes from Xenopus and human has been analyzed. The Groucho, TLE and ESG proteins all share a similar structure, consisting of a conserved amino-terminal domain, a variable middle region, and highly conserved carboxyl-terminal WD-40 repeats. The Drosophila Groucho transcriptional corepressor protein has been shown to interact with the DNA-binding bHLH domain of Enhancer of split, Hairy and Deadpan proteins, which proteins are involved in neurogenesis, segmentation and sex determination, respectively. Human TLE1 protein has been demonstrated to interact with mammalian AML1 protein, which regulates hematopoiesis and osteoblast differentiation. The AES proteins from human, mouse, rat and Xenopus exhibit strong similarity to the amino-terminal domain of Groucho proteins; however, the biological function remains to be elucidated. PMID- 10809081 TI - Postoperative administration of interleukin-12 significantly enhances the anti tumor immune response of MBT-2 bladder cancer bearing mice. AB - This study, using the MBT-2 murine bladder tumor model, mainly investigated the role of interleukin-12 (IL-12) in the specific antitumor immune response of a tumor-bearing host when systemically administrated after surgery. Day 17 tumor bearing mice (D17TBM) along with non-tumor bearing naive mice were treated with daily intraperitoneal (i.p.) injection of IL-12 (0.25 microg/mouse) from day 18 to day 24 for a total of 7 doses. Their splenocytes were obtained on Day 31 for natural killer cells (NK), lymphokine activated killer cells (LAK) and cytotoxic T lymphocyte (CTL) activity assay and lymphocyte subsets phenotypic analysis. The tumor suppression effect of systemic IL-12 administration was evaluated based on the tumor outgrowth of the higher number of tumor cells rechallenged 24 hours after resectioning of the primary tumor. After evaluation on Day 31, the result of in vitro cytotoxicity assay revealed that systemic administration of IL-12 mainly enhanced the splenic LAK and CTL activities in non-tumor-primed naive mice, and the NK activity in tumor-primed D17TBM, respectively. However, in vivo administration of IL-12 with or without IL-2 failed to upgrade the proportions of either CD4+ CD44+ or CD8+ CD44+ T cells subsets in the spleens and regional inguinal lymph nodes (LNs) of both the D17TBM and naive mice. However, the splenic CD8+ CD44+ T-cell subset in the IL-12-treated D17TBM increased prominently after further culturing in the presence of IL-2 400 units/ml plus IL 12 25 ng/ml for 4 days. The fact that systemic administration of IL-12 significantly suppressed the outgrowth of Day-18 challenged tumor cells, especially in D17TBM, clearly indicates that the established specific antitumor immunity in tumor-primed D17TBM was efficiently augmented. From the results of this study, we conclude that, after surgical resection of a primary tumor, systemic administration of IL-12 can be an effective adjuvant therapy because it demonstrates a significant augmentation effect on the tumor-specific immune response in the tumor-primed host. PMID- 10809082 TI - Association of HLA-te22 antigen with anti-nuclear antibodies in Chinese patients with erosive oral lichen planus. AB - The purpose of this study was to determine the frequencies of the presence of serum anti-nuclear antibodies (ANA) and smooth muscle antibodies (SMA) in 76 patients with oral lichen planus (OLP), in 77 patients with other oral mucosal diseases, and in 41 healthy control subjects. HLA phenotypes in some of the patients with OLP and recurrent aphthous ulcers (RAU) were determined to show whether there was an association of HLA antigens with the presence of autoantibodies. Indirect immunofluorescence techniques with mouse liver or stomach as the substrate were used to detect the serum ANA or SMA, respectively. The B lymphocytes isolated from peripheral blood were used for HLA typing by means of a standard microcytotoxicity assay. We found that the positive rate of serum ANA in patients with OLP (29%, p < 0.01), especially in patients with erosive OLP (34%, p < 0.001), was significantly higher than that in the normal control subjects (5%). The frequency of serum SMA in patients with OLP (20%, p < 0.01), in patients with RAU (17%, p < 0.01), or in patients with oral squamous cell carcinomas (41%, p < 0.001) was also significantly higher than that in normal control individuals (0%). In the erosive OLP group, the HLA-Te22 antigen occurred more frequently in patients with positive ANA (75%, p < 0.05) than in those with negative ANA (25%). We conclude that there is an association of HLA Te22 antigen with ANA in Chinese patients with erosive OLP. PMID- 10809083 TI - Milk progesterone profiles in various reproductive states in dairy buffaloes under field conditions. AB - Fifty-one dairy buffaloes in the last two months of gestation were selected at seven private peri-urban farms in the Peshawar district. Observations were recorded in buffaloes during normal (NBS, August to January) and low breeding seasons (LBS, February to July). After parturition, rectal examination of reproductive organs was carried out. Estrus detection was made through visual observation and the use of intact bull. Postpartum ovulation was confirmed by ovarian palpation per rectum and milk progesterone levels (MPL), determined through radio-immunoassay. MPL was higher (p < 0.01) at various intervals in NBS calves (1.97 +/- 0.30 ng/ml) as compared to LBS calves (0.68 +/- 0.08 ng/ml). During LBS, MPL remained < 0.30 ng/ml up to the third fortnight and started rising later, reaching a peak of 1.27 ng/ml during the sixth fortnight. During NBS, there was a sharp rise in MPL during the second fortnight, reaching 3.64 ng/ml during the sixth fortnight. MPL was significantly different on different experimental farms (p < 0.01). MPL reached the lowest levels on the day of estrus (0.10 ng/ml), reached it's peak on day 7 and started declining on day 17 of estrus. MPL showed two postpartum elevations. In true anestrus buffaloes, MPL remained consistently low. However, in the anestrus period, silent ovulations were also noted, as reflected by increasing MPL without estrus signs. In pregnant buffaloes, MPL remained > 1 ng/ml. Results of the study showed that the low postpartum reproductive performance in dairy buffaloes during LBS was primarily due to inadequate functioning of the corpus luteum in secreting optimum concentrations of progesterone. The higher incidence of silent estrus during LBS indicated improved management for the detection of estrus. PMID- 10809084 TI - Distribution of CAG repeat size in the dentatorubral and pallidoluysian atrophy (DRPLA) gene in a normal population in Taiwan. AB - Dentatorubral and pallidoluysian atrophy (DRPLA) is an autosomal dominant neurodegenerative disorder with expansion of trinucleotide CAG repeats in the coding region of the gene. Expansion of the repeat tract beyond the normal range produces gene products with extended polyglutamine tracts. In this study, we analyzed the distribution of the CAG repeats in the DRPLA alleles in a normal Taiwanese population. We observed 15 different alleles and found that the range of the CAG repeat number was from 7-21. The most frequent allele contained 15 CAG repeats that represented 20% of the total analyzed alleles, followed by the 17 repeats (15.8%). The heterozygosity rate of this locus was 88%. Twelve parents-to children transmissions of the DRPLA alleles in a Machado-Joseph disease family appeared to be normal without any alteration of the CAG repeat numbers. Phenotypes of DRPLA overlapped those of autosomal dominant cerebellar ataxia (ADCA). In order to identify DRPLA patients in Taiwan, we screened six autosomal dominant cerebellar ataxia patients without expansion in known spinocerebellar ataxia genes. All six patients had the repeat numbers within the normal range; thus, the possibility of DRPLA could be excluded. PMID- 10809085 TI - Astrocytes modulate thapsigargin-induced changes in calcium concentration and neuronal survival. AB - When mature cerebellar granule neurons (CGN) grown in high K+ (25 mM K+, HK) serum containing medium are subjected to the HK/serum deprivation, they are destined for neuronal death. In this study, we attempted to elucidate the roles of endoplasmic reticular (ER) Ca2+-store and co-cultured astrocytes in HK/serum deprivation induced neuronal death. Thapsigargin (TG), an inhibitor of ER Ca2+ ATPase was simultaneously applied with normal K+ (5 mM K+, NK) serum free medium, and its effects on neuronal death in either astrocyte-poor or astrocyterich culture were examined. By means of the fura-2 microfluorimetric technique, we monitored the changes of the intracellular Ca2+ concentration, [Ca2+]i, associated with neuronal death under various treatments. The results obtained showed that in astrocyte-poor cultures of mature CGN (10 days in vitro, DIV), the basal level of [Ca2+]i markedly decreased from 184 +/- 5 to 89.7 +/- 5 nM 24 h after HK/serum deprivation. Although treatment with TG slightly increased the [Ca2+]i to 117.6 +/- 4 nM, the survival rate of the neurons was even worse; it was reduced from 49 +/- 4% to 28 +/- 2%. In the astrocyte-rich cultures, HK/serum deprivation also caused a profound reduction of neuronal [Ca2+]i, from 166 +/- 3 to 90.2 +/- 6 nM, accompanied by even more serious neuronal death (95.5 +/- 1%). On the other hand, treatment with TG in astrocyterich cultures further lowered the [Ca2+]i to 65 +/- 2 nM but markedly improved the neuronal survival rate from 4.5 +/- 1% to 60 +/- 2% in a concentration-dependent manner. The strong implication of these findings is that ER Ca2+-store and astrocytes participate in modulating the responses of neurons to stress stimulation. PMID- 10809086 TI - Cellular mechanisms for diminished scarring with aging. AB - The study of an age-dependent spectrum of scar formation is driven by the desire to understand and recapitulate scarless healing. Although focus in the past has been directed toward scarring in the fetus, less exuberant scarring is a common clinical observation in the elderly. Cell turnover is a major contributor to the development of scar tissue and is governed by the proliferative and apoptotic cellular fractions within a healing wound. We hypothesize that the balance between cell proliferation and apoptosis during late stages of excisional wound healing is, at least in part, responsible for age-related variations in scarring potential. Full-thickness 7-mm ulcers (four per ear), exposing bare cartilage, were made on the inner surface of the ear on 12 young and 12 aged New Zealand White rabbits. Analyses were performed at days 15, 21, and 28 postwounding. A previously described Scar Elevation Index was derived from histomorphometric analysis, along with the quantification of epithelial ingrowth and total cellularity. Apoptotic cellular fractions were derived from TdT-mediated dUTP nick end-labeling assay-stained histologic sections; proliferative fractions were derived from proliferating cell nuclear antigen-labeled serial sections. Young rabbits demonstrated significantly greater scar elevation/area. Apoptosis was strongly associated with progress of epithelialization in both groups. Significantly higher proliferative indices were seen in the young and were sustained through day 28, by which time levels had substantially declined in the aged. No differences in apoptotic indices were demonstrated between groups at any time point. The clinical observation of less exuberant scarring with aging is supported by this animal model. Apoptosis follows the progression of epithelialization but does not appear to independently influence scar morphology. A diminished proliferative response during later stages of healing is an important contributing mechanism for the decrease in scar formation seen in the elderly. PMID- 10809087 TI - Long-term outcome of cleft lip nasal reconstruction in childhood. AB - This study documents the pattern of unilateral cleft lip nasal reconstruction in the practice of one surgeon at a tertiary cleft center, evaluating the long-term appearance outcome of single-operation unilateral cleft lip nasal reconstruction in childhood. A retrospective medical record review was performed for all patients with a diagnosis of unilateral cleft lip and age greater than 15 years. Operative notes were reviewed, recording 15 variables identifying specific rhinoplastic maneuvers. Nasal appearance outcome analysis was performed for all patients who underwent only one nasal surgery before 12 years of age (n = 19). Standard frontal whole face photographic images were presented as opposing pairs in a looseleaf binder to two panels, 1 of 10 lay persons and 1 of 10 plastic surgeons. Each pair consisted of photographs of the same patient at different ages in one of three combinations: preoperative-perioperative, perioperative longest postoperative, or preoperative-longest postoperative. Participants were asked to compare the appearance of the noses in the two photographs and assign a rating based on a 5-point Likert scale. Statistical analyses were performed on the data collected in the aesthetic analysis. The effect of surgery upon nasal appearance was assessed by comparing the preoperative and perioperative photographs. The effect of growth was assessed by comparing the perioperative and long-term postoperative photographs. The combined effect of surgery and growth was assessed by comparing the preoperative and long-term postoperative photographs. The data were assessed by lay and professional evaluators, together and separately, to determine whether differences existed. The majority of patients did not undergo revisional nasal surgery, whereas those who did usually had one nasal operation. Most revisional nasal surgery was performed in conjunction with other cleft-related secondary surgery. A majority of lay and professional evaluators perceived revisional nasal surgery as improving nasal appearance in the short-term and to a lesser degree in the long-term, as compared with the preoperative state. Evaluations of revisional nasal surgery are generally constant between the short-term and long-term postoperative images. Lay evaluations may be contaminated by a general decline in attractiveness with aging. Patient preference should be a major factor in the decision for nasal revision surgery. Multiple means of assessing nasal appearance outcome need to be used to validate results. Nasal appearance outcomes need to be correlated with outcomes with respect to nasal morphology and function as well as patient and parent satisfaction. PMID- 10809088 TI - Designs for correction and camouflage of bilateral clefts of the lip and palate. AB - The bilateral cleft anomaly is difficult to correct and camouflage because of the double lack of many important landmarks and the shortness of skin in the midvertical plane. A possible solution in patients who have some columella or in those of races not needing a long columella is the strap flap advancement of the nostril sills and alar bases. In all other cases, the forked flap is the method of choice for adequate correction and camouflage of the bilateral cleft lip-nose deformity. PMID- 10809089 TI - Nasal reconstruction using both cranial bone and ear cartilage. AB - In this article, an anatomic reconstruction is described using cranial bone graft for the bony part of the nose while reconstructing the cartilaginous part with ear cartilage. This modification provides protection of the nose from the traumatic forces creating a flexible nasal tip. A modification is described and compared with the nasal reconstruction by calvarial bone itself. PMID- 10809090 TI - Quality of life and disease-specific functional status following microvascular reconstruction for advanced (T3 and T4) oropharyngeal cancers. AB - In an effort to evaluate quality-of-life benefits of ablative head and neck cancer surgery and microvascular reconstruction, a longitudinal study was undertaken in which patients with T3 or T4 oropharyngeal cancers without systemic metastases at presentation were administered both general and disease-specific quality-of-life instruments preoperatively and postoperatively. In an initial prospective pilot study, 17 cancer patients were evaluated both preoperatively and postoperatively using the Medical Outcomes Short-Form Health Survey questionnaire (SF-36) and the Performance Status Scale for Head and Neck Cancer Patients. In the second part of the study, the need was recognized for a different disease-specific measure, for more frequent intervals of longitudinal follow-up (rather than be limited by a single data collection point), and for a noncancer control group. Since then, 17 more cancer patients were evaluated in the second part of the study and were compared with patients who had similar reconstructions after suffering head and neck trauma and also with age-matched controls. Instead of the performance status scale, the University of Washington Head and Neck Quality of Life questionnaire was substituted. Interval assessments were done at 1, 3, 6, and 12 months and preoperatively. Whereas many of the general and disease-specific quality of life subclasses initially worsened following extensive surgery and radiation therapy, most returned to the preoperative baseline by 6 months following conclusion of treatment and surpassed pretreatment values at 1 year. It can be concluded, based on this study, that large resections and reconstructions for head and neck cancer patients are justified in terms of outcome; the resection controls the local disease, and the microvascular reconstruction restores quality of life and functional status. PMID- 10809091 TI - Healing of burns after treatment with 670-nanometer low-power laser light. AB - Recent investigations have reported contradictory results on the influence of low power laser light on wound healing. Low-power laser with a power output of 250 mW and an emitted laser light of 670 nm have been insufficiently investigated to date. The effect of a 250-mW/670-nm laser light on the healing of burning wounds in rats was investigated. Thirty rats were burned on both flanks. One wound was irradiated with 670-nm laser light (2 J/cm2), whereas the other side remained untreated. Macroscopic evaluation of the wounds was performed daily; 10, 20, and 30 days after burning, 10 rats were killed and the wounds histologically evaluated. Neither macroscopic nor histologic examination of the irradiated wound showed accelerated wound healing when compared with control wounds. In the present study, irradiation of burns with a 250-mW/670-nm laser light produced no beneficial effects on wound-healing processes. PMID- 10809092 TI - Effect of obesity on flap and donor-site complications in free transverse rectus abdominis myocutaneous flap breast reconstruction. AB - The purpose of this study was to assess the effect of obesity on flap and donor site complications in patients undergoing free transverse rectus abdominis myocutaneous (TRAM) flap breast reconstruction. All patients undergoing breast reconstruction with free TRAM flaps at our institution from February 1, 1989, through May 31, 1998, were reviewed. Patients were divided into three groups based on their body mass index: normal (body mass index <25), overweight (body mass index 25 to 29), obese (body mass index > or =30). Flap and donor-site complications in the three groups were compared. A total of 936 breast reconstructions with free TRAM flaps were performed in 718 patients. There were 442 (61.6 percent) normal-weight, 212 (29.5 percent) overweight, and 64 (8.9 percent) obese patients. Flap complications occurred in 222 of 936 flaps (23.7 percent). Compared with normal-weight patients, obese patients had a significantly higher rate of overall flap complications (39.1 versus 20.4 percent; p = 0.001), total flap loss (3.2 versus 0 percent; p = 0.001), flap seroma (10.9 versus 3.2 percent; p = 0.004), and mastectomy flap necrosis (21.9 versus 6.6 percent; p = 0.001). Similarly, overweight patients had a significantly higher rate of overall flap complications (27.8 versus 20.4 percent; p = 0.033), total flap loss (1.9 versus 0 percent p = 0.004), flap hematoma (0 versus 3.2 percent; p = 0.007), and mastectomy flap necrosis (15.1 versus 6.6 percent; p = 0.001) compared with normal-weight patients. Donor-site complications occurred in 106 of 718 patients (14.8 percent). Compared with normal-weight patients, obese patients had a significantly higher rate of overall donor-site complications (23.4 versus 11.1 percent; p = 0.005), infection (4.7 versus 0.5 percent; p = 0.016), seroma (9.4 versus 0.9 percent; p <0.001), and hernia (6.3 versus 1.6 percent; p = 0.039). Similarly, overweight patients had a significantly higher rate of overall donor-site complications (19.8 versus 11.1 percent; p = 0.003), infection (2.4 versus 0.5 percent; p = 0.039), bulge (5.2 versus 1.8 percent; p = 0.016), and hernia (4.3 versus 1.6 percent; p = 0.039) compared with normal-weight patients. There were no significant differences in age distribution, smoking history, or comorbid conditions among the three groups of patients. Obese patients, however, had a significantly higher incidence of preoperative radiotherapy and preoperative chemotherapy than did patients in the other two groups. A total of 23.4 percent of obese patients had preoperative radiation therapy compared with 12.3 percent of overweight patients and 12.4 percent of normal-weight patients; 34.4 percent of obese patients had preoperative chemotherapy compared with 24.5 percent of overweight patients and 17.7 percent of normal-weight patients. Multiple logistic regression analysis was used to determine the risk factors for flap and donor-site complications while simultaneously controlling for potential confounding factors, including the incidence of preoperative chemotherapy and radiotherapy. In summary, obese and overweight patients undergoing breast reconstruction with free TRAM flaps had significantly higher total flap loss, flap hematoma, flap seroma, mastectomy skin flap necrosis, donor-site infection, donor-site seroma, and hernia compared with normal-weight patients. There were no significant differences in the rate of partial flap loss, vessel thrombosis, fat necrosis, abdominal flap necrosis, or umbilical necrosis between any of the groups. The majority of overweight and even obese patients who undertake breast reconstruction with free TRAM flaps complete the reconstruction successfully. Both such patients and surgeons, however, must clearly understand that the risk of failure and complications is higher than in normal-weight patients. Patients who are morbidly obese are at very high risk of failure and complications and should avoid any type of TRAM flap breast reconstruction. PMID- 10809093 TI - Innervated island flaps in morphofunctional vulvar reconstruction. AB - In this article, the authors present their own experience in vulvar reconstruction following vulvectomy using two different innervated island flaps according to the size and site of the defect. Island-flap mobilization is possible thanks to the rich blood supply of the perineal region. The methods described are a "V-Y amplified sliding flap from the pubis" and a "fasciocutaneous island flap" raised from one or both gluteal folds. The V-Y amplified sliding flap from the pubis is indicated when the defect is symmetric and located anteriorly. This flap is harvested from the pubis and vascularized by the deep arterial network of the pubis. Sensory innervation is provided by branches of the ileo-inguinal nerve. The fasciocutaneous island flap, raised from one or both gluteal folds, can be used following hemivulvectomy or radical vulvectomy, respectively, to cover posteriorly located defects. Vascularization is provided by the musculocutaneous perforating branches of the pudendal artery, whereas sensory innervation is maintained through the perineal branches of the pudendal nerve. Twenty-two patients have undergone reconstructive surgery of the vulvar region from 1989 to date. On 14 patients, a V-Y amplified sliding flap was used; on 7 patients, reconstruction was carried out by island flaps raised from the gluteal fold. Both techniques are compatible with inguino-femoral lymphadenectomy, and they allow for a correct morphofunctional reconstruction and provide good local sensibility. The final result is aesthetically satisfactory, as all final scars are hidden in natural folds. PMID- 10809094 TI - Correction of pincer-nail deformity using dermal grafting. AB - Pincer-nail syndrome has been described as distortion in the shape of the nails with excessive transverse curvature of the plate that increases from proximal to distal, leading to pinching and loss of soft tissue in the affected digit, resulting in severe pain. Many treatments have been recommended, but an effective long-term method that preserves the nail matrix has not been described. A method of dermal grafting under the nail matrix is described, and the results of treatment of six digits are reported. Five women and one man with an average age of 52 were treated. The affected digit was the thumb in four patients and the great toe in two patients. Follow-up averaged 25 1/2 months. The results were good in all cases with only one side of one nail remaining slightly curved. Pain was relieved in all cases, and complete adherence of the new nail plate occurred. Dermal grafting seems to provide excellent long-term treatment of the pincer-nail deformity with preservation of the nail matrix. PMID- 10809095 TI - The incidence of recurrence after endoscopic carpal tunnel release. AB - Endoscopic carpal tunnel release has been used to decompress the median nerve in carpal tunnel syndrome for over the past decade, with an advantage (over the traditional "open" release) being decreased pain in the postoperative period. The goals of this study were to attempt to define the recurrence rate after endoscopic carpal tunnel release and to determine if it differs from that of open technique. The charts of 191 consecutive carpal tunnel syndrome patients treated operatively at the University of Missouri were reviewed. For this study, recurrent carpal tunnel syndrome was defined as documented cases in which the symptoms had resolved following surgical release but subsequently recurred, requiring surgical rerelease of the carpal tunnel. All endoscopic releases were performed using the Chow two-portal technique. Statistical analysis was performed using Fisher's exact test. A total of 103 patient hands had open carpal tunnel releases; 88 were endoscopically released. Total follow-up time (from the initial release) averaged 29 months for the open group and 22 months for the endoscopic group. There were no recurrences in the open group and six recurrences in the endoscopic group (7 percent, p = 0.008). All six recurrences were in worker's compensation patients. The median time between endoscopic release and rerelease was 8.5 months. There seems to be a statistically higher incidence of recurrence of carpal tunnel syndrome after endoscopic release compared with the traditional "open" release in our cases. Although the pathogenesis of this increased rate of recurrence is not clear, this should be considered when planning surgical release of the volar carpal ligament for carpal tunnel syndrome. PMID- 10809096 TI - Fingertip reconstructions using partial-toe transfers. AB - Fifty-six partial toes were transferred to reconstruct fingertip deficits. The transfers from the big toe mainly consisted of 3 trimmed big toetips, 3 vascularized nail grafts, 3 onychocutaneous flaps, 19 thin osteo-onychocutaneous flaps, and 2 hemipulp flaps. The transfers from the second toe mainly consisted of 8 trimmed second toetips, 5 reduced second toes, and 9 whole distal phalanges. The average values of postoperative sensory recovery of the osteo-onychocutaneous flaps including the vascularized nail grafts were 3.1 (Semmes-Weinstein test) and 6.3 mm (moving two-point discrimination) at 2.6 years after the transfer; those of the thin osteo-onychocutaneous flaps were 3.1 and 7.2 mm at 2.0 years after surgery; those of the trimmed big toe tip transfers were 3.61 and 6.5 mm at 1.8 years after surgery; and those of the trimmed second toetip transfers were 3.37 and 6.3 mm at 2.6 years after transfer. Those of the distal phalanx of the second toe were 3.41 and 7.9 mm at 1.2 years after surgery, and those of the reduced second toe were 3.2 and 6.7 mm at 10.6 months after surgery. PMID- 10809097 TI - Delayed toe transplantation: experimental study and clinical application. AB - Delay of the operation in toe transplantation was performed with circulatory crisis, and 100 percent survival of the transplants was achieved. In 10 cases of toe transplantation in which circulatory crisis occurred, the operation was delayed. The vascular pedicle was divided, and the toe was transferred 17 to 21 hours after the donor toe was dissected. To investigate the mechanism of delayed toe transplantation, experiments in rabbits were carried out. In 36 rabbits, the hind limbs were severed except for the vascular pedicle. The endothelin content and nitric oxide content of the tissues in the hind limb and the arterial wall were tested in postoperative intervals of 4, 8, 16, 24, and 48 hours, respectively. The results showed that in 10 cases of delayed toe transplantation, all toes survived. As for the results of the experiment, there was significant increase of endothelin content and decrease of nitric oxide content in the local tissues and the arterial wall 4 to 8 hours after the operation. In postoperative 16 to 24 hours, endothelin content returned to normal level, and nitric oxide content increased remarkably. It is concluded that delayed pedicle division and toe transfer is an effective method in toe transplantation with vascular variations and circulatory crisis. The mechanism is related to the endothelin and nitric oxide content in the local tissue and the arterial wall. The indications for delayed toe transfer are also discussed. PMID- 10809098 TI - Strategic considerations on the configuration of free flaps and their vascular pedicles combined with Ilizarov distraction in the lower extremity. AB - For the injury of the lower leg associated with both bone and soft-tissue defect, the combined free flap and the Ilizarov distraction method were described as a useful treatment modality. During the procedure of distraction, however, revisions were frequently needed to change the pin position or to change the flap configuration. In case of flap ischemia, distraction should be delayed or abandoned. Then, a vascularized bone transfer might be necessary. To avoid these complications and achieve safe distraction, the configuration of the flap with its vascular pedicle should be carefully planned in terms of the future bony lengthening procedures and the concomitant soft-tissue changes of the lower leg. According to the response of local tissue to the distraction process, the lower limb can be divided into four compartments (active mobile, passive mobile, receptive, and restrictive). The configuration of the transferred free flap with its vascular pedicle can be classified into five types. To minimize the undue forces to the vascular pedicle and reduce the possibility of vascular compromise, the transferred free flap should have the configuration that its vascular pedicle lies in the territory of the mobile compartment. In performing free-tissue transfer combined with the Ilizarov method in the lower extremity, the configuration of the flap with its vascular pedicle should be carefully planned, and the characteristics of lower leg tissue should be kept in mind during the distraction. PMID- 10809099 TI - Palatal distraction in a canine cleft palate model. AB - The purpose of this study was to determine whether the canine hard palate can be lengthened by distraction osteogenesis in a cleft palate model using a mostly submucosal distractor. Five mongrel dogs were used. After raising mucoperiosteal flaps, a midline strip of bone was removed from the hard palate of each dog to simulate the bony defect seen in a cleft palate. A transverse osteotomy was then made to separate the posterior segment of the hard palate from the anterior segment. Posterior osteotomies were also made laterally parallel to the teeth so that the 2 posterior segments (one on either side of the bony cleft) were mobile. An intraoral distractor that was mostly submucosal was attached to the anterior hard palate and both segments of the mobilized posterior hard palate. Radiopaque bone markers were placed, and x-rays were obtained. After a 10-day latency period, the distractor was expanded 0.675 mm per day until it had been lengthened 10.125 mm. Distractors were left in place for an additional 8 weeks. After distractor removal, animals were observed for an additional 8 weeks before euthanization. Follow-up x-rays and histologic examinations were performed. New bone formation was found at the site of distraction in all dogs at the time of death. This new bone was seen on the follow-up x-rays and on histologic examination of the hard palates using both hematoxylin and eosin staining and Masson's trichrome stain. Distraction osteogenesis using a mostly submucosal device is an effective technique for lengthening the hard palate in a canine cleft palate model. The technique may eventually provide an alternative treatment for velopharyngeal incompetence in humans that is more precise and involves less morbidity than existing treatments. PMID- 10809100 TI - Prolonged survival of musculoskeletal xenografts with combined cyclosporine and 15-deoxyspergualin. AB - This study was undertaken to evaluate the feasibility of performing vascularized musculoskeletal xenografts between mice and rats using immunosuppression. Vascularized musculoskeletal grafts were harvested from the hind limb of C57BL/6J (B6) mice, transplanted heterotopically into Lewis rats, and revascularized by microanastomoses of the graft artery and the recipient femoral artery and the graft vein to the recipient femoral vein. Recipient rats were divided into four groups. Group 1 received no immunosuppression (n = 10), group 2 was treated with cyclosporine (10 mg/kg/day; n = 10), group 3 was treated with 15-deoxyspergualin (5 mg/kg/day; n = 10), and group 4 received both cyclosporine and 15 deoxyspergualin (n = 10). Graft survival was directly examined on postoperative days 4, 7, and 14. In vitro assays were performed using mixed lymphocyte reactions and anti-donor cytotoxic antibody assays to assess the recipient's immune response. Grafts were examined by histology and immunohistochemistry. All grafts in group 1 were rejected by day 4. In groups 2 and 3, all grafts were rejected by day 7. In group 4, however, 8 of 10 recipients had viable grafts on day 14. Data from mixed lymphocyte reactions showed that cell-mediated immune responses were uniformly suppressed in groups 2, 3, and 4 compared with group 1. However, anti-donor antibody production was only partly suppressed in groups 2 and 3, suggesting that graft rejection was primarily caused by circulating cytotoxic anti-donor antibodies in groups 1, 2, and 3. Histologic observations in groups 1, 2, and 3 confirmed the important role of the humoral mechanism in xenograft rejection. Furthermore, immunohistochemical results demonstrated that the small vessels in the rejected grafts showed anti-rat immunoglobulin and complement depositions. Only a combination therapy of cyclosporine and 15 deoxyspergualin attenuated the rejection of xenografts. PMID- 10809101 TI - A functional model for whole limb transplantation in the rat. AB - To develop a functional model for the study of whole limb transplantation, inbred Lewis rats were used as both donors and recipients. In this model, the recipient biceps femoris muscle was elevated from its distal attachment to preserve part of the adductor function of the limb after surgery. The tibial, peroneal, and sural branches of the sciatic nerve were anastomosed separately to provide faster and more precise functional recovery. For control sensory evaluation, the saphenous branches of the femoral nerve were not reattached. A flat intramedullary pin stabilized with methyl methacrylate was used to rigidly immobilize the femur. The transplanted limbs started bearing weight at 17 to 22 days. Walking on the plantar surface of the hock and adduction of the toes gradually decreased, and the rats developed a normal walking pattern. Sciatic and tibial function indexes, based on walking track analysis, correlated well with clinical observations. In this study, a new model for limb transplantation was developed that provided good and reliable sensory and ambulatory recovery. PMID- 10809102 TI - Increased free fat-graft survival with the long-term, local delivery of insulin, insulin-like growth factor-I, and basic fibroblast growth factor by PLGA/PEG microspheres. AB - The present investigation evaluates the effects of long-term, local delivery of insulin, insulin-like growth factor-1 (IGF-1), and basic fibroblast growth factor (bFGF) on fat-graft survival using a poly (lactic-co-glycolic-acid)-polyethylene glycol (PLGA/PEG) microsphere delivery system. Twelve-micrometer PLGA/PEG microspheres incorporated separately with insulin, IGF-1, and bFGF were manufactured using a double-emulsion solvent-extraction technique. Inguinal fat from Sprague Dawley rats was harvested, diced, washed, and mixed with (1) insulin microspheres, (2) insulin-like growth factor-1 microspheres, (3) basic fibroblast growth factor microspheres, (4) a combination of the insulin and IGF-1 microspheres, and (5) a combination of insulin, IGF-1, and bFGF microspheres. The treated fat grafts were implanted autologously into subdermal pockets in six animals for each group. Animals receiving untreated fat grafts and fat grafts treated with blank microspheres constituted two external control groups (six animals per external control group). At 12 weeks, all fat-graft groups were compared on the basis of weight maintenance and a histomorphometric analysis of adipocyte area percentage, indices of volume retention and cell composition, respectively. Weight maintenance was defined as the final graft weight as a percent of the implanted graft weight. All growth factor treatments significantly increased fat-graft weight maintenance objectively, and volume maintenance grossly, in comparison with the untreated and blank microsphere-treated controls. Treatment with insulin and IGF-1, alone or in combination, was found to increase the adipocyte area percentage in comparison with fat grafts treated with bFGF alone or in combination with other growth factors. In conclusion, the findings of this study indicate that long-term, local delivery of growth factors with PLGA/PEG microspheres has the potential to increase fat-graft survival rates. Further, the type of growth factor delivered may influence the cellular/stromal composition of the grafted tissue. PMID- 10809103 TI - De novo adipose tissue generation through long-term, local delivery of insulin and insulin-like growth factor-1 by PLGA/PEG microspheres in an in vivo rat model: a novel concept and capability. AB - This study was undertaken to characterize the duration of long-term growth factor delivery by poly(lactic-co-glycolic-acid)-polyethylene glycol (PLGA/PEG) microspheres and to evaluate the potential of long-term delivery of insulin and insulin-like growth factor-1 (IGF-1) for the de novo generation of adipose tissue in vivo. PLGA/PEG microspheres containing insulin and IGF-1, separately, were produced by a double-emulsion solvent-extraction technique. In the first phase of the experiment, the in vitro release kinetics of the microspheres were evaluated for the optical density and polyacrylamide gel electrophoresis of solutions incubated with insulin-containing microspheres for four different periods of time (n = 1). The finding of increased concentrations of soluble insulin with increased incubation time confirmed continual protein release. In the second stage of the experiment, 16 rats were divided equally into four study groups (insulin, IGF-1, insulin + IGF-1, and blank microspheres) (n = 4). Insulin and IGF-1 containing microspheres were administered directly to the deep muscular fascia of the rat abdominal wall to evaluate the potential for de novo adipose tissue generation via adipogenic differentiation from native nonadipocyte cell pools in vivo. Animals treated with blank microspheres served as an external control group. At the 4-week harvest period, multiple ectopic islands of adipose tissue were observed on the abdominal wall of the animals treated with insulin, IGF-1, and insulin + IGF-1 microspheres. Such islands were not seen in the blank microsphere group. Hematoxylin and eosin-stained sections of the growth factor groups demonstrated mature adipocytes interspersed with fibrous tissue superficial to the abdominal wall musculature and continuous with the fascia. Oil Red-O stained sections demonstrated that these cells contained lipid. Computer aided image analysis of histologic sections confirmed that there were statistically significant increases in the amount of "ectopic" adipose neotissue developed on the abdominal wall of animals treated with growth factor microspheres. In conclusion, this study confirms the long-term release of proteins from PLGA/PEG microspheres up to 4 weeks and demonstrates the potential of long-term local insulin and IGF-1 to induce adipogenic differentiation to mature lipid-containing adipocytes from nonadipocyte cell pools in vivo at 4 weeks. PMID- 10809104 TI - Facial contour restoration in Barraquer-Simons syndrome using two free anterolateral thigh flaps. AB - Barraquer-Simons syndrome, or cephalothoracic lipodystrophy, is characterized by fat atrophy of an obscure pathogenesis involving the face and, eventually, the thoracic region. Simultaneously, fat hypertrophy of the lower extremities, a nephropathy, and complement anomalies may be observed. We presented two patients with the typical features of this disease, as well as a previously undescribed vascular and perivascular inflammation of the facial arteries and veins that caused problems with microvascular anastomosis. Both patients were treated with a bilateral transfer of the anterolateral thigh flap, which has not been reported previously. In contrast to other transfers previously reported, the fat tissue of this flap is never affected by the disease and is redundantly present. Placing the fascia of the flaps toward the skin allows for strong fixation to the temporal region and guarantees a stable result with a smooth facial contour. PMID- 10809105 TI - Cranial reconstruction for metastatic breast cancer. AB - All women with advanced breast cancer who are medically stable despite their disease are candidates for tumor extirpation and reconstruction. Advanced breast cancer today is incurable, and many prognostic factors can be used to try to predict a clinical course and response to therapy; however, no guidelines are available. Our case report most likely represents a metastasis to the calvarium with intracranial extension, reported to occur in about 3 percent of primary breast cancer patients. As demonstrated here, tumor ablation with immediate, one stage reconstruction of large scalp defects is possible without the need for free tissue transfer or a delay in adjuvant therapy. Local tissue rearrangement has been employed for coverage of defects up to 50 percent of the cranium. The resulting donor defects can be closed with split-thickness skin grafts over pericranium. Serial tissue expansion and rearrangement can be used secondarily to replace skin grafts with hair-bearing scalp. Bony defects can be managed with either autogenous or alloplastic materials. Split-calvarial bone grafts can be harvested from the same operative field and cover small to medium-sized defects. Other sources of autogenous grafts include split ribs and iliac bone. Metals, calcium ceramics, and polymers such as methylmethacrylate can be used to cover intracranial contents and restore calvarial contour when defects are large or when autogenous material is not available. Palliation from tumor burden, prevention of pathologic fracture and oncologic emergencies, controlling pain, and enhancing quality of life are the goals of the oncologic and reconstructive surgeons in cases of advanced breast cancer. These goals are becoming even more important as new forms and combinations of chemotherapy, radiation, and gene therapy are extending the life expectancy of women with breast carcinoma. PMID- 10809106 TI - Closure of hemicorporectomy with bilateral subtotal thigh flaps. AB - Hemicorporectomy is typically performed with a circumferential truncal incision, and the wound is closed primarily. Wound disruption is a common complication, especially at the base of the wound closure and posteriorly at the lumbar vertebral level. We report a case of the use of bilateral subtotal thigh flaps for the closure of a hemicorporectomy wound in a patient with a defect extending up to the high lumbar region. The subtotal thigh flap is a well-vascularized thick flap that provides a firm support for the abdominal viscera and is a large flap that can be used to close even a high lumbar defect. PMID- 10809107 TI - Congenital synostosis of the fourth and fifth metacarpals treated by free bone grafting from the fusion site. AB - Simultaneous correction of the alignment and the shortening of the deformed little finger in congenital synostosis of the fourth and fifth metacarpals was accomplished by insertion of a wedge-shaped bone block harvested from the fusion site following transverse osteotomy of the fifth metacarpal. Correction can be performed simply and successfully without morbidity of the donor site when this technique is applied to suitable patients. PMID- 10809108 TI - A practical method of macrophotography in microsurgery. AB - A simple method to take macrophotographs that show details even in the microsurgical field was discussed. The method uses any single lens up to 80 mm or any zoom lens in the reverse position with a single-lens reflex camera. In this manner, high reproduction ratios larger than the actual size of the object can be achieved. PMID- 10809109 TI - Reconstruction of the upper portion of the ear by using an ascending helix free flap from the opposite ear. AB - Reconstruction of partial, marginal defects of the ear has been a challenge. The ascending helix free flap based on superficial temporal vessels has been described and used solely to repair nose defects. We used reversed pedicle helical free flap for the repair of a major loss of the upper one-third of the opposite auricle. The method permits the transfer of tissue of the same quality with satisfactory cosmetic result. The equalization of the ears in dimension was accomplished with minimal donor-site deformity. PMID- 10809110 TI - Extended lower trapezius island myocutaneous flap: a fasciomyocutaneous flap based on the dorsal scapular artery. AB - The lower trapezius island myocutaneous flap is a useful flap in head and neck reconstruction. It is thin and pliable and can reach defects in most areas of the head and neck. Its usefulness in head and neck reconstruction has often been limited or discouraged by reports of significant failure rates. In this study, the vascular anatomy and clinical use of the extended lower trapezius myocutaneous flap based solely on the dorsal scapular artery system are elucidated, and experience gained performing 20 flaps over the past 5 years by using the extended lower trapezius myocutaneous flap is reported. The vascular anatomy of the dorsal scapular artery system is reviewed in 13 fresh cadaveric dissections by using methylene blue, latex injection studies, and radiologic examination. PMID- 10809111 TI - Endoscopic tenotomy of the sternocleidomastoid muscle: new method for surgical correction of muscular torticollis. PMID- 10809112 TI - Wall Street's growing influence on plastic surgery. AB - The advent of managed care has unleashed market forces on the health care system. One result of these new pressures is a shift from nonprofit to Wall Street-based financing. This report quantifies these trends by comparing health organizations' financial structures in the 1980s and now. The reasons behind this shift and the function of the stock market are examined. A review of Wall Street's key financial measures confirms that health care has shifted to the stock market as its principal means of financing. The stock market works by assigning a current price to a company's stock based on estimates for future earnings. Thus, companies desire predictability in their costs, revenues, and profits. Plastic surgeons can master this system by meeting the challenges imposed by Wall Street financing. Important steps include continuously measuring costs and outcomes of procedures, demanding cost data from hospitals and payers, using these data to improve costs and outcomes, and taking advantage of the system's openness to innovation and easier access to capital. As they seek to protect their role as medical decision makers under the new free-market system, plastic surgeons can benefit from understanding the mechanisms of the stock market. PMID- 10809113 TI - Current therapy of cutaneous melanoma. AB - Melanoma is a growing public health problem. Optimal care of the melanoma patient is multidisciplinary, but plastic surgeons and other surgical specialties play a central role in the management of these patients. Although surgery remains the mainstay of therapy for melanoma, several recent clinical studies have helped to clarify the biology of the disease and have changed the patterns of care for patients with melanoma. The advent of lymphatic mapping for interrogation of regional lymph nodes and interferon as the first effective postsurgical adjuvant therapy have had a major impact on the care of melanoma in the United States and elsewhere. This article will review the current clinical approach and therapy for cutaneous melanoma. The diagnosis, prognostic variables, staging evaluation, current surgical and medical treatment, and follow-up guidelines for patients with all stages of melanoma are reviewed. Recent studies, controversies, and directions of future investigational therapies will be discussed. PMID- 10809114 TI - The Socratic method in plastic surgery education: a lost art revisited. PMID- 10809115 TI - The lateral osteotomy in rhinoplasty: clinical and radiographic rationale for osteotome selection. AB - A preferred osteotome for endonasal osteotomy would facilitate reliable, complete osteotomies with minimal soft-tissue trauma. In this report, a radiographic evaluation of the bony lateral nasal wall thickness along the track of a high-low high osteotomy was undertaken to guide the determination of appropriate osteotome size. Bone window axial computed tomographic scans were evaluated in 56 patients with a mean age of 48 years (range, 19 to 86 years). The average thickness along the site of lateral osteotomy was determined to be 2.47 mm (standard deviation, 0.47) in male patients and 2.29 mm (standard deviation, 0.40) in female patients. On the basis of these data, clinical evaluation of prototype 3- and 2.5-mm low profile guarded osteotomes was undertaken in comparison with a "standard" 4-mm low-profile guarded osteotome to assess both their reliability and the degree of intranasal trauma, as reflected by intranasal mucosal tears. Although 2- and 3-mm unguarded osteotomes are time-tested, they may be reliable only in the hands of the most experienced surgeons. Therefore, a low-profile guard was included in the osteotome design to allow the surgeon to engage the bone securely and minimize the risk of slippage. Forty patients underwent rhinoplasty, for a total of 80 lateral osteotomies; the mean age of the patients was 38 (range, 16 to 75). In all cases, lateral osteotomies were accomplished with one pass. The 4-mm osteotome causes intranasal mucosal tears in 95 percent of osteotomies, the 3-mm osteotome in 34 percent, and the 2.5-mm in 4 percent. Early postoperative edema and ecchymosis were comparable among the groups. One patient, who underwent osteotomies with a 4-mm osteotome, had excessive postoperative narrowing, possibly due to his wearing of eyeglasses earlier than directed. This report suggests that proper selection of osteotome and attention to proper surgical technique results in a reliable, minimally traumatic lateral osteotomy through the endonasal approach. The 2.5-mm osteotome was reliable and the least traumatic to soft tissue of the osteotomes evaluated. PMID- 10809116 TI - Rhinoplasty: personal evolution and milestones. AB - Over the past 35 years, aesthetic rhinoplasty has evolved from a generic, reductive operation to a highly individualized, problem-specific operation that often combines augmentation with reduction. The author's experience has been marked by the following conceptual and technical milestones that have contributed to an ongoing exploration and advancement of nasal surgery: (1) vestibular stenosis: diagnosis of a surgical consequence; (2) etiology and treatment of supratip deformity: the dynamic relationship of soft-tissue contour to skeleton; (3) etiology and treatment of the tip with inadequate projection: tip graft design; (4) practical aesthetics of balance: the augmentation-reduction approach to rhinoplasty; (5) support of the middle vault: functional and aesthetic effects; (6) malposition of the lateral crura: recognition and management; and (7) the significance of the middle crura: clinical and aesthetic considerations. PMID- 10809117 TI - Brachioplasty technique with the use of molds. AB - Over previous years many surgical techniques have been developed to correct lipodystrophy and brachial skin laxity while providing a natural, discreet contour and symmetrical scars, a goal that has not always been achieved. In this study, the authors classify the alterations of the arm into three degrees and propose a different surgical treatment, to be used alone or associated with another procedure, for each classification. They developed a new technique that uses a mold to mark the incision in an italic double S-shape. From 1996 to 1998, 20 patients who underwent this surgery showed symmetrical and smaller scars with better results and minimal complications. PMID- 10809118 TI - Immediate hair transplantation into a newly closed wound to conceal the final scar on the hair-bearing skin. AB - A surgical incision after suturing usually leaves a visible scar on the hair bearing skin, even after optimal wound conditions. The conspicuousness of such a scar results from its linear continuity and hairlessness. To prevent this effect, a row of micrografts or minigrafts was inserted between the wound edges immediately after wound closure. The hair grafts that were transplanted were dissected from the discharged skin in the same surgical procedure, if feasible. Otherwise, a mini donor strip was harvested from the mastoid scalp to dissect the hair grafts. The final linear scar was interrupted and concealed sufficiently with the growth of the transplanted hairs. Tension-free closure is required to obtain a satisfactory result with this technique. PMID- 10809119 TI - Anchor epicanthoplasty combined with out-fold type double eyelidplasty for Asians: do we have to make an additional scar to correct the Asian epicanthal fold? AB - The epicanthal fold along with a lack of a superior palpebral fold, excessive fat, and laxity of pretarsal skin represent the ethnic characteristics and a traditional sense of beauty in the Asian upper eyelid. But, too prominent an epicanthal fold may ruin an otherwise beautiful eye; furthermore, it becomes a restriction that makes the out-fold type double eyelidplasty, one of the two main types of double eyelidplasty, impossible. If a double eyelid as an out-fold type is desired, a concomitant epicanthoplasty should be performed with the possibility of hypertrophic scarring of the medial canthal area in Asians. To address the Asian epicanthal fold without danger of hypertrophic scarring, the authors developed an anchor epicanthoplasty technique that leaves no additional scar when combined with a double eyelidplasty. This technique is based on the concept of trimming of muscle and soft tissue under the Asian epicanthal fold and downward medial advancement and anchoring of the medial canthal skin to the deep tissue. The technique consists of five procedures based on the assumed causes of the Asian epicanthal fold: (1) augmentation rhinoplasty, (2) downward medial advancement of the medial upper lid skin, (3) removal of the superficial insertion of the medial canthal ligament and selective removal of the orbicularis oculi muscle, (4) subcutaneous contouring of the thick nasal skin, and (5) anchoring of the medial end of the incision to the deep tissue. During the past 12 years (1988 to 1999), 67 anchor epicanthoplasty procedures have been performed. Twenty-eight cases were followed up for more than 3 months, and all of the patients were satisfied with the results. There were only a few minor complications, which could be corrected with minimal revision. As an ancillary procedure to a double eyelidplasty, this anchor epicanthoplasty can reduce the Asian epicanthal fold and make a double fold as an out-fold type without an additional scar. In terms of hypertrophic scarring and compatibility with out fold type double eyelidplasty, this anchor epicanthoplasty is the best method for correcting Asian epicanthal fold compared with other preexisting procedures. Other advantages of this technique are a wide range of applications and no compromise of medial, canthal skin to interfere with other epicanthoplasty techniques. Some disadvantages of this technique are technical difficulty and the possibility of active bleeding. PMID- 10809120 TI - A national survey of complications associated with suction lipectomy: what we did then and what we do now. PMID- 10809121 TI - Collegiality: another casualty of the marketplace. PMID- 10809122 TI - Floating proboscis. PMID- 10809123 TI - The use of transfixion mattress sutures to stabilize spreader grafts. PMID- 10809124 TI - An unusual complication of microtia repair. PMID- 10809125 TI - The plastic syringe as soft-tissue protector during drainage tube insertion. PMID- 10809126 TI - Effective delayed brush treatment of an extensive traumatic tattoo. PMID- 10809127 TI - The use of an island scalp flap based on the superficial temporal artery for the reconstruction of the sideburn. PMID- 10809128 TI - The findings of magnetic resonance imaging in Pierre Robin syndrome. PMID- 10809129 TI - Supraomohyoid cellulitis from placement of a dental implant. PMID- 10809130 TI - Bone planning for the radial osteocutaneous flap. PMID- 10809131 TI - The partial double-barrel free vascularized fibular graft: a solution for long mandibular defects. PMID- 10809132 TI - Subpectoral implants in weight lifters. PMID- 10809133 TI - Operating on patients with Ehlers-Danlos syndrome. PMID- 10809134 TI - The replacement of a degloved digit using a plastic bag. PMID- 10809135 TI - An unusual foot lipoma. PMID- 10809136 TI - A simple technique for fibrin glue application in skin grafting. PMID- 10809137 TI - Scientific approach to presenting and summarizing data. PMID- 10809138 TI - Varaztad H. Kazanjian, D.M.D., M.D. PMID- 10809139 TI - Beware of the low-pressure bed substitute. PMID- 10809140 TI - Failure to remove soft tissue injected with liquid silicone with the use of suction and honesty in scientific medical reports. PMID- 10809141 TI - Hypothesis: naked plasmid DNA is taken up by cells in vivo by a receptor-mediated process. AB - Following the initial demonstration that intramuscularly-injected naked plasmid DNA (pDNA) is expressed in myofibers, it was shown that pDNA can be used for vaccination purposes. More recent studies have indicated that naked pDNA can also achieve high levels of transgene expression in vivo. This efficiency of naked pDNA expression, especially via intravascular route, is truly astounding. In this prospective review, we examine the possible mechanisms of naked pDNA uptake. The possible mechanisms; (a) large membrane disruption, (b) small membrane pores, and (c) receptor-mediated endocytosis, are considered in turn. Some recent original laboratory data relevant to these hypotheses are also presented. PMID- 10809142 TI - Control of transgene expression using local hyperthermia in combination with a heat-sensitive promoter. AB - BACKGROUND: Local production of therapeutic proteins, e.g. for cancer treatments, is based on gene therapy approaches and requires tight spatial and temporal control of gene expression. Here we demonstrate the use of local hyperthermia of varying intensity and duration to control the expression of a transgene under control of the thermoinducible hsp70 (heat shock protein) promoter. METHODS: Heat induced expression of the EGFP (green fluorescent protein) reporter gene was characterized using a stably transfected glioma C6 cell line expressing the EGFP gene under control of the heat inducible minimal hsp70 promoter both in vitro and in vivo for subcutaneous tumors in immunodeficient mice. RESULTS: A heat shock of 20-30 min at 43 degrees C in cell culture led to a maximum EGFP concentration at about 24 h. Heat treatments at higher temperature (up to 48 degrees C) but with shorter durations (down to 30 s) also induced strong EGFP expression. Local heating in situ led to gradients in EGFP expression which decreased with increasing distance from the heat source. CONCLUSION: Local hyperthermia, in combination with a heat sensitive promoter, represents a method for the spatial and temporal control of transgene expression. PMID- 10809143 TI - Pro-apoptotic treatment with an adenovirus encoding Bax enhances the effect of chemotherapy in ovarian cancer. AB - BACKGROUND: Tumor cell heterogeneity and resistance to chemotherapy-mediated cell death are major obstacles in cancer therapy. It has been reported that expression of the pro-apoptotic molecule Bax can induce cell death or sensitize tumor cells to chemotherapy in stable cell clones derived from tumor cells. However, these studies are limited in that they cannot represent the heterogeneity of cancer cells observed in vivo. In this study, we have further explored the therapeutic potential of Bax. METHODS: Using an inducible recombinant Bax adenovirus, we screened a panel of ovarian cancer cell lines and primary patient-derived ovarian tumor cells for their sensitivity to Bax-mediated cytotoxicity. Apoptotic cell death was evaluated qualitatively with Hoechst staining and quantitatively with MTS and Annexin V-based assays. Endogenous levels of both Bcl-2 and Bax protein and p53 status were evaluated. The potential of bax to sensitize ovarian cancer lines to chemotherapy was also tested. Dose-response curves were generated to evaluate cell death. RESULTS: Overexpression of Bax directly induced apoptosis in both ovarian cancer cell lines and the patient-derived primary cancer cells. However, the sensitivity of these cells to Bax varied and appeared to be independent of both the status of p53 and the endogenous levels of bcl-2 or Bax, critical molecules in the apoptotic pathway. Importantly, overexpression of Bax significantly enhanced chemotherapy-induced cytotoxicity in both established cell lines and primary ovarian carcinoma cells. CONCLUSIONS: These studies suggest that overexpression of Bax alone or in combination with chemotherapy may provide a means to overcome the problems imposed by the heterogeneous nature of tumors, ultimately augmenting the efficacy of chemotherapy in patients suffering from ovarian cancer. PMID- 10809144 TI - A regulatory network for the efficient control of transgene expression. AB - BACKGROUND: Expression of heterologous genes in mammalian cells or organisms for therapeutic or experimental purposes often requires tight control of transgene expression. Specifically, the following criteria should be met: no background gene activity in the off-state, high gene expression in the on-state, regulated expression over an extended period, and multiple switching between on- and off states. METHODS: Here, we describe a genetic switch system for controlled transgene transcription using chimeric repressor and activator proteins functioning in a novel regulatory network. In the off-state, the target transgene is actively silenced by a chimeric protein consisting of multimerized eukaryotic transcriptional repression domains fused to the DNA-binding tetracycline repressor. In the on-state, the inducer drug doxycycline affects both the derepression of the target gene promoter and activation by the GAL4-VP16 transactivator, which in turn is under the control of an autoregulatory feedback loop. RESULTS: The hallmark of this new system is the efficient transgene silencing in the off-state, as demonstrated by the tightly controlled expression of the highly cytotoxic diphtheria toxin A gene. Addition of the inducer drug allows robust activation of transgene expression. In stably transfected cells, this control is still observed after months of repeated cycling between the repressed and activated states of the target genes. CONCLUSIONS: This system permits tight long-term regulation when stably introduced into cell lines. The underlying principles of this network system should have general applications in biotechnology and gene therapy. PMID- 10809145 TI - Overexpression of Mad transcription factor inhibits proliferation of cultured human hepatocellular carcinoma cells along with tumor formation in immunodeficient animals. AB - BACKGROUND: Dominant negative regulation of critical cell cycle molecules could perturb survival of cancer cells and help develop novel therapies. METHODS: To perturb the activity of c-Myc, which regulates G0/G1 transitions, we overexpressed Mad1 protein with an adenoviral vector, AdMad. Studies were conducted with established cell lines, including HepG2, HuH-7 and PLC/PRF/5 liver cancer cells, RAT-1A embryonic fibroblasts and U373MG astrocytoma cells. RESULTS: After AdMad-treatment, transduced cells exhibited decreased proliferation rates in culture conditions. RAT-1A embryonic fibroblasts and U373MG astrocytoma cells showed accumulations in G0/G1, whereas HepG2 and HuH-7 cells accumulated in G0/G1, and additionally in G2/M, albeit to a lesser extent. An in vitro assay using hepatocyte growth factor to stimulate proliferation in HuH-7 cells showed blunting of growth factor responsiveness, along with inhibition of cell cycle progression in AdMad-treated cells. No cytotoxicity was observed in AdMad-treated cells in culture, although cells lost clonogenic capacity in soft agar. In vivo assays using HepG2 cell tumors in immunodeficient mice showed that overexpression of AdMad prevented tumorigenesis. CONCLUSIONS: These studies indicate roles of Mad in G2/M, as well as the potential of manipulating cell cycle controls for treating liver cancer. PMID- 10809146 TI - Systemic linear polyethylenimine (L-PEI)-mediated gene delivery in the mouse. AB - BACKGROUND: Several nonviral vectors including linear polyethylenimine (L-PEI) confer a pronounced lung tropism to plasmid DNA when injected into the mouse tail vein in a nonionic solution. METHODS: and results We have optimized this route by injecting 50 microg DNA with excess L-PEI (PEI nitrogen/DNA phosphate = 10) in a large volume of 5% glucose (0.4 ml). In these conditions, 1-5% of lung cells were transfected (corresponding to 2 ng luciferase/mg protein), the other organs remaining essentially refractory to transfection (1-10 pg luciferase/mg protein). beta-Galactosidase histochemistry confirmed alveolar cells, including pneumocytes, to be the main target, thus leading to the puzzling observation that the lung microvasculature must be permeable to cationic L-PEI/DNA particles of ca 60 nm. A smaller injected volume, premixing of the complexes with autologous mouse serum, as well as removal of excess free L-PEI, all severely decreased transgene expression in the lung. Arterial or portal vein delivery did not increase transgene expression in other organs. CONCLUSIONS: These observations suggest that effective lung transfection primarily depends on the injection conditions: the large nonionic glucose bolus prevents aggregation as well as mixing of the cationic complexes and excess free L-PEI with blood. This may favour vascular leakage in the region where the vasculature is dense and fragile, i.e. around the lung alveoli. Cationic particles can thus reach the epithelium from the basolateral side where their receptors (heparan sulphate proteoglycans) are abundant. PMID- 10809147 TI - Induction of anti-tumor immunity by intrasplenic administration of a carcinoembryonic antigen DNA vaccine. AB - BACKGROUND: We have previously reported that intramuscular, intradermal or epidermal gene gun administration of a plasmid encoding carcinoembryonic antigen (CEA) under transcriptional regulatory control of the cytomegalovirus (CMV) early promoter/enhancer elicits CEA-specific humoral and cellular immune responses in mice with resultant immunoprotection against challenge with syngeneic, CEA expressing colon adenocarcinoma cells. METHODS: In the present work, we examine the ability of this DNA vaccine construct (pCEA) to elicit CEA-specific immunity following intrasplenic administration. Groups of mice were immunized with pCEA by intrasplenic or intramuscular injection. Six weeks later, mice were evaluated for the presence of anti-CEA humoral responses and were challenged with syngeneic, CEA-expressing colon carcinoma cells. RESULTS: Intrasplenic administration of pCEA produced a frequency of CEA-specific antibody responses comparable to that elicited by intramuscular pCEA inoculation. Both intrasplenic and intramuscular administration of pCEA generated IgG2a antibody responses to CEA, consistent with the induction of T helper-1-biased immune responses. In addition, partial immunoprotection against tumor challenge was observed after a single plasmid DNA dose by either route of administration. Subsequent studies revealed that antibody responses to intrasplenic DNA vaccination are dose and schedule dependent. CONCLUSION: These findings support future investigations of DNA vaccination strategies that specifically promote the uptake of plasmid by splenocytes. PMID- 10809148 TI - Cancer gene therapy--new approaches to tumour cell killing. PMID- 10809162 TI - Future trends in colorectal cancer research. Conclusions from the workshop. PMID- 10809163 TI - Dexamethasone modulates the activity of the eel branchial Na+/K+ATPase in both chloride and pavement cells. AB - In fish, gills actively accumulate ions in freshwater (FW) with Na+ absorption taking place at the level of pavement cells, and excrete monovalent ions, mainly Na+ and Cl-, through the chloride cells in sea water (SW). The Na+/K+ATPase plays a crucial role in the functionality of osmoregulatory cells and we showed previously that angiotensin II modulates its activity in the eel gill (1). We here show the effects of synthetic steroid dexamethasone (DEX) on the activity of Na+/K+ATPase in both gill pavement and chloride cells from FW- and SW-adapted animals. Results showed that in the chloride cells 100 nM DEX provoked a significant increment in the activity of Na+/K+ATPase in both SW- and FW-adapted animals. This effect was greatest at 2 hours in SW, and at 6 hours in FW. The increment in the activity of the Na+/K+ATPase was dose-dependent in both environmental adaptations. Conversely, in pavement cells from FW-adapted eels 100 nM DEX decremented the activity of Na+/K+ATPase (4-fold reduction after 6 hour incubations), while in SW, DEX increased the enzyme activity of about 25% at 2 hours, and of about 55% at 6 hours. These results are consistent with the different physiological roles that pavement and chloride cells have in the two different adaptive conditions. PMID- 10809164 TI - Probing the size of a hydrophobic binding pocket within the allosteric site of muscarinic acetylcholine M2-receptors. AB - Hexane-bisammonium-type compounds containing lateral phthalimide moieties are known to have a rather high affinity for the allosteric site of muscarinic M2 receptors. In order to get more insight into the contribution of the lateral substituents for alloster binding affinity, a series of compounds with unilaterally varying imide substituents were synthesized and tested for their ability to retard allosterically the dissociation of [3H]N-methylscopolamine from the receptor protein (control t1/2 = 2 min; 3 mM MgHCO4, 50 mM Tris, pH 7.3, 37 degrees C). Among the test compounds, the naphthalimide containing agent (half maximum effect at ECs5,diss = 60 nM) revealed the highest potency. Apparently, its affinity for the allosteric site in NMS-occupied receptors is 20fold higher compared with the phthalimide containing parent compound W 84. Analysis of quantitative structure-activity relationships yielded a parabolic correlation between the volume of the lateral substituents and the allosteric potency. The maximal volume was determined to be approximately 600 A3 suggesting that the allosteric binding site contains a binding pocket of a defined size for the imide moiety. PMID- 10809165 TI - Elevated hepatic apolipoprotein A-I transcription is associated with diet-induced hyperalphalipoproteinemia in rabbits. AB - Past studies have shown that a high saturated fatty acid diet containing coconut oil elevates plasma HDL cholesterol and apolipoprotein A-I (apoA-1) in rabbits through a mechanism involving increased synthesis. We have extended those studies by investigating expression of the hepatic apolipoprotein A-I gene and other lipid related genes in that model. Rabbits fed a diet containing 14% coconut oil for 4 weeks showed HDL-C elevations of 170% to 250% over chow-fed controls with peak differences occurring at 1 week. Plasma apoA-I levels were also increased over this time frame (160% to 180%) reflecting the HDL-C changes. After 4 weeks, there were no differences in plasma VLDL-C or LDL-C levels in chow versus coconut oil-fed rabbits. Hepatic levels of apoA-I mRNA in coconut oil-fed animals were elevated 150% after 4 weeks compared to chow-fed controls; hepatic mRNA levels for ten other genes either decreased slightly (apoB, LCAT, hepatic lipase, albumin, ACAT, and HMG CoA reductase) or were unchanged (CETP, apoE, LDL receptor, and acyl CoA oxidase). Nuclear run-on transcription assays revealed that coconut oil feeding for 4 weeks caused a 220% increase in hepatic apoA-I transcription rate compared to controls; no change was observed for CETP and apoE. Treatment of cultured rabbit liver cells with various saturated fatty acids and sera from chow-fed and coconut oil-fed rabbits did not alter apoA-I mRNA levels as observed in vivo. These data demonstrate that coconut oil elevates plasma HDL-C and apoA-I by increasing hepatic apoA-I transcription while expression of other genes involved in lipid metabolism are reduced or unchanged in response to coconut oil feeding. PMID- 10809166 TI - Laryngeal C-fiber afferents are not involved in the apneic response to laryngeal wood smoke in anesthetized rats. AB - Laryngeal exposure to wood smoke in rats evokes a reflex apnea which is mediated through superior laryngeal afferents (J. Appl. Physiol. 83: 723-730, 1997). To study the role of laryngeal C-fiber afferents in eliciting this response, capsaicin aerosol (0.05 - 0.2 microg/ml) and 5 ml of wood smoke were delivered separately into a functionally isolated larynx of anesthetized Sprague-Dawley rats at a constant flow rate of 1.4 ml/s, while animals breathed spontaneously. Studies were repeated after either an intravenous injection of ruthenium red (2 mg/kg; n = 8), a perineural capsaicin treatment (200 microg/ml for 5 min; n = 8) of the superior laryngeal nerves, or a perineural sham treatment (n = 8); Ruthenium red inhibits the stimulation of afferent C-fiber nerve endings by capsaicin, whereas perineural capsaicin treatment selective blocks the conduction of C-fiber afferents. Either ruthenium red or perineural capsaicin treatment abolished the apneic response to laryngeal capsaicin, but did not significantly affect the apneic response to laryngeal wood smoke. Furthermore, the apneic responses to both types of irritants were not significantly altered by perineural sham treatment, yet were completely eliminated by a subsequent denervation of superior laryngeal nerves. Our results suggest that superior laryngeal C-fiber afferents are not involved in eliciting the reflex apneic response to laryngeal wood smoke in anesthetized rats. It is speculated that this response may result mainly from the stimulation of myelinated afferents, possibly laryngeal irritant receptors. PMID- 10809167 TI - Investigation on the mechanism involved in the effects of agmatine on ethanol induced gastric mucosal injury in rats. AB - Effects of agmatine, an endogenous metabolite formed by decarboxylation of L arginine, on ethanol-induced gastric mucosal injury were investigated in rats. Agmatine at 1 and 10 mg/kg i.p doses significantly increased ethanol-induced gastric mucosal injury. This effect of agmatine was abolished completely by pretreatment with idazoxan, an imidazoline receptor-antagonist and alpha2 receptor- antagonist, (0.5 mg/kg i.p), partly by yohimbine, an alpha2 receptor- antagonist, (1 mg/kg i.p) but not by L-arginine, a precursor of nitric oxide, (500 mg/kg i.p). Our results suggest that agmatine had a potent ulcerogenic effect mediated, at least in part, by both alpha2-adrenoceptors and imidazoline receptors. PMID- 10809168 TI - Telomerase activity is not related to apoptosis in leukemic cell lines. AB - Any deregulation of apoptosis or an escape from cellular senescence will drive the cells to neoplasia. It remains unclear whether there is a direct linkage between apoptosis and telomerase activity particularly in transformed cell lines. In the present study, we investigated the telomerase activities in three leukemic cell lines (HL-60, U937 and K562) after treating these cells with various doses of antitumor drugs, puromycin or actinomycin D (ActD). Our results showed that HL 60 cells underwent apoptosis rapidly when treated with either 20 microg/ml of puromycin or 5 microg/ml of Act D with more than 60% of the cells becoming apoptotic at 6 hrs and almost 100% at 12 hrs. But telomerase activity analyzed by TRAP assay in these apoptotic cells remained unchanged as compared with the untreated control cells suggesting that whether the cells were apoptotic or not, it had no effect on telomerase activity. However, if lower dosages of the drugs were used, that is, 0.5-1.5 microg/ml of puromycin or 0.01-0.5 microg/ml of Act D, a decrease in telomerase activity was observed at 24-48 hrs, and was completely undetectable at 72 hrs. This decrease in telomerase activity was dose- and time-dependent. The suppression of telomerase activity by low doses of these two drugs is probably due to the inhibitory effect of the drugs on protein translation or RNA transcription rather than direct inhibition of the telomerase activity. Flow cytometry analysis of the cell cycle of the drug-treated cells showed that these drugs unselectively induced apoptosis at all phases of the cell cycle and was unrelated to the changes in telomerase activity. Similar results were observed in U937 and K562 cells except that K562 cells underwent apoptosis more slowly than the former two cell lines. PMID- 10809169 TI - Morphine treatment affects the regulation of high mobility group I-type chromosomal phosphoproteins in C6 glioma cells. AB - High mobility group (HMG) I-type chromosomal phosphoproteins HMG I/Y and HMG I-C were investigated following morphine treatment of C6BU-1 glioma. Cells were labelled with [32P]-orthophosphoric acid. Electrophoretic profiles and autoradiograms of the control cells revealed the presence of HMG I and HMG I-C proteins. HMG Y was not detected. Northern blot analysis showed a single HMG I/Y transcript. Treatment with morphine lowered the [32P]-incorporation in HMG I and HMGI-C proteins and the level of the HMG I/Y transcript. However, it did not change the protein ratios on the Coomassie stained gels. These results suggest that morphine may trigger independent reaction pathways affecting either transcription regulation and/or postsynthetic phosphorylation of the preexisting HMG I-type proteins. In addition, opposing changes in the postsynthetic phosphorylation of HMG 14 and histones H1AB were also noticed. PMID- 10809170 TI - Nuclear all-trans retinoic acid receptor status in rat liver: a comparison of effects of three different stressors--immobilization, laparotomy, and 2-deoxy-D glucose. AB - Retinoic acids and their cognate nuclear receptors exert a substantial regulatory role in cell growth and development as well as in the neuroendocrine system. These effects are primarily mediated by all-trans retinoic acid receptors (RARs), members of the steroid/thyroid hormone receptor superfamily of ligand inducible transcription factors. The present study was undertaken in order to compare the effects of immobilization stress (IMO), laparotomy, and 2-deoxy-D-glucose (2DG) induced intracellular glucopenia on both nuclear RAR affinity and concentration in the rat liver. IMO when compared to non-stressed group of rats, significantly reduced the RAR maximal binding capacity (Bmax) in liver, with the equilibrium association constant (Ka) remaining unchanged. No significant changes of the RAR Bmax and the Ka, were observed in liver of rats that underwent laparotomy. In contrast, a single dose of 2DG (500 mg/kg) resulted in a significant increase of the RAR Bmax 10 h after 2DG application, with the Ka remaining unchanged. Shorter intervals, 1 or 5 h after 2DG application were ineffective on both the RAR Bmax and Ka. In the 2DG-adapted rats (6 doses of 2 DG, 500 mg/kg; 1 dose/day), decapitated 24 h after the last 2DG dose, the RAR Bmax was found significantly higher when compared to control group of animals. No further effect of the next dose of 2DG to repeatedly injected rats on the RAR Bmax and Ka was observed in animals killed 5 h after the seventh dose of 2DG. 2DG-induced intracellular glucopenia markedly up-regulates RARs in liver, but does not change the affinity of the receptor. Thus, the effect of 2DG on RAR concentration in liver specifically differs from that of IMO or laparotomy. PMID- 10809171 TI - Progesterone receptor isoforms expression pattern in the rat brain during the estrous cycle. AB - Progesterone receptor (PR) isoforms expression was determined in the hypothalamus, the preoptic area, the hippocampus and the frontal cerebral cortex of the rat at 12:00 h on each day of the estrous cycle by using reverse transcription coupled to polymerase chain reaction. Rats under a 14:10 h light dark cycle, with lights on at 06:00 h were used. We found that PR-B isoform was predominant in the hypothalamus, the preoptic area and the frontal cerebral cortex. Both PR isoforms were similarly expressed in the hippocampus. The highest PR-B expression was found on proestrus day in the hypothalamus; on metestrus in the preoptic area; and on diestrus in the frontal cortex. We observed no changes in PR isoforms expression in the hippocampus during the estrous cycle. These results indicate that PR isoforms expression is differentially regulated during the estrous cycle in distinct brain regions and that PR-B may be involved in progesterone actions upon the hypothalamus, the preoptic area and the frontal cortex of the rat. PMID- 10809172 TI - Mechanisms of nordihydroguaiaretic acid-induced [Ca2+]i increases in MDCK cells. AB - The effect of nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor, on Ca2+ signaling in Madin Darby canine kidney (MDCK) cells has been investigated. NDGA (10-100 microM) increased [Ca2+]i concentration-dependently. The [Ca2+]i increase comprised an initial slow rise and a plateau over a time period of 5 min. Ca2+ removal partly inhibited the Ca2+ signals induced by 25-100 microM NDGA and abolished that induced by 10 microM NDGA. In Ca(2+)-free medium, pretreatment with 0.1 mM NDGA for 12 min abolished the [Ca2+]i increase induced by the mitochondrial uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP; 2 microM) and the endoplasmic reticulum (ER) Ca2+ pump inhibitor thapsigargin (1 microM). However, 0.1 mM NDGA still increased [Ca2+]i after Ca2+ stores had been depleted by pretreating with 2 microM CCCP, 1 microM thapsigargin and 0.1 mM cyclopiazonic acid. NDGA (50 microM) activated Mn2+ quench of fura-2 fluorescence at 360 nm excitation wavelength, which was almost abolished by 50 microM La3+. This implies NDGA induced Ca2+ influx mainly via a La(3+)-sensitive pathway. Consistently, 50 microM La3+ pretreatment inhibited 0.1 mM NDGA-induced [Ca2+]i increase. Adding 3 mM Ca2+ increased [Ca2+]i in cells pretreated with 0.1 mM NDGA in Ca(2+)-free medium, suggesting NDGA activated capacitative Ca2+ entry. Pretreatment with 0.1 mM NDGA for 200 s prior to Ca2+ did not alter 1 microM thapsigargin-induced capacitative Ca2+ entry. Pretreatment with 40 microM aristolochic acid to inhibit phospholipase A2 reduced 0.1 mM NDGA-induced Ca2+ release by 65%, but inhibiting phospholipase C with 2 microM U73122 had little effect. This suggests NDGA induced Ca2+ release was independent of inositol 1,4,5-trisphosphate (IP3), but was modulated by phospholipase A2. PMID- 10809173 TI - Changes in urine 8-hydroxydeoxyguanosine levels of super-marathon runners during a four-day race period. AB - We have determined the urinary 8-hydroxydeoxyguanosine (8-OHdG) levels of five well trained supra-marathon runners during a four-day race. The daily running distances of the four-day race were the following; 93 km, 120 km, 56 km and 59 km, respectively. Pre-race and post-race urine samples were collected on each day and analyzed by a monoclonal antibody technique. The urinary 8-OHdG content increased significantly on the first day and tended to decrease from the third day. By the fourth day 8-OHdG content was significantly less than measured on the first three days. The serum creatine kinase activity changed in a similar fashion, showing a large increase (P<0.001) up to the third day when it decreased significantly from the peak value (P<0.05). We conclude that extreme physical exercise causes oxidative DNA damage to well trained athletes. However, repeated extreme exercise-induced oxidative stress does not propagate on increase of urinary 8-OHdG, but rather causes an adaptation leading to normalization of oxidative DNA damage. PMID- 10809174 TI - Increased apoptosis in U937 cells over-expressing lipocortin 1 (annexin I). AB - The potential involvement of endogenous lipocortin 1 in the process of cellular apoptosis, particularly in cells of the myelo-monocytic lineage, has been investigated. U937 cells were transfected either with an antisense or a sense DNA for lipocortin 1 and the stable clones 36.4AS clone (20-40% lower lipocortin 1 levels) and 15S (30% higher lipocortin 1 levels) were obtained. Cell apoptosis was induced by incubation with tumor necrosis factor-alpha: optimal responses were observed within a 24 h incubation period at a 5 ng/ml concentration. Apoptosis was assessed both morphologically, by annexin V binding and cell cycle analysis with propidium iodide. Whilst no consistent difference was seen between wild type cells and clone 36.4AS, a higher incidence of apoptosis (ranging from +30% to + 60%) was observed in the 15S clone. Release of arachidonic acid from loaded cells was promoted by 24 h incubation with the cytokine, and a higher degree of release was measured in the 15S clone. These data indicate that endogenous intracellular lipocortin 1 is involved in the promotion of apoptosis in cells of the myelo-monocytic derivation. PMID- 10809175 TI - Role of regucalcin in calcium signaling. AB - Regucalcin was discovered in 1978 as a calcium-binding protein that does not contain EF-hand motif of Ca(2+)-binding domain [M. Yamaguchi and T. Yamamoto, Chem. Pharm. Bull. 26 1915-1918 (1978)]. In recent years, regucalcin has been demonstrated to play an important role as a regulatory protein in Ca2+ signaling in rat liver and kidney cells. The organization of the rat regucalcin gene consists of seven exons and six introns. The mRNA is mainly present in liver and kidney with a size of 1.8 kb. Hepatic regucalcin mRNA expression has been shown to be stimulated by various factors including calcium, calcitonin, insulin, and estrogen in rats. The mRNA is also expressed in hepatoma cells (Morris hepatoma, HepG2, and rat hepatoma H4-II-E cells). Regucalcin plays a role in the maintenance of intracellular Ca2+ homeostasis due to activating Ca2+ pump enzymes in the plasma membrane (basolateral membrane) and microsomes of liver and renal cortex cells. Moreover, regucalcin has an inhibitory effect on the activation of Ca2+/calmodulin-dependent enzymes and protein kinase C. Also, regucalcin has been demonstrated to regulate nuclear function in liver cells; it can inhibit Ca(2+) activated DNA fragmentation, DNA and RNA synthesis, protein kinase and protein phosphatase activities in the nuclei. Such an effect is also seen in the nuclei of regenerating rat liver. Regucalcin may play a physiological role in the control for overexpression of proliferative cells. Regucalcin has been proposed to be an important regulatory protein in Ca2+ signaling system, and it plays a multifunctional role in liver and kidney cells. PMID- 10809176 TI - Adenosine and its receptor agonists regulate nitric oxide production and RAW 264.7 macrophages via both receptor binding and its downstream metabolites inosine. AB - Adenosine and its receptor agonists enhanced the production of nitric oxide (NO) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The enhancement of LPS induced NO production by adenosine, as represented by the amount of its oxidation products, nitrite and nitrate, was inhibited by adenosine uptake inhibitors, such as dipyridamole, S(4-nitrobenzyl)-6-thioinosine (NBTI) and S(4-nitrobenzyl)-6 thioguanosine (NBTG). These indicate that the uptake of adenosine by macrophages is a prerequisite for the enhancement effects observed. A downstream metabolite of adenosine, inosine, also potentiated the LPS-induced NO production in a dose dependent manner while its enhancement effect was also inhibited by dipyridamole. However, the degree of enhancement by inosine on NO production and nitric oxide synthase (NOS) activity in LPS-treated RAW 264.7 was weaker than the effect of adenosine. Furthermore, adenosine agonists also enhanced the NO production in a dose-dependent manner, but were not specific for A1, A2 nor A3 adenosine receptor. Adenosine uptake inhibitors had no effects on the enhancement activity of the adenosine receptor agonists. Thus, extracellular receptor/s may also play an important role in the observed enhancement responses. The results of this study indicate that the enhancement effects of adenosine on NO production in macrophages could be mediated by the extracellular adenosine receptors as well as the downstream metabolites of adenosine. PMID- 10809177 TI - A simplified isolated perfused rat liver apparatus: characterization and measurement of extraction ratios of selected compounds. AB - A simplified isolated perfused rat liver (IPRL) preparation has been developed and evaluated. The liver is briefly perfused in situ prior to being placed into a 37 degrees C oven and suspended from a stand. This set-up takes about 5 min. A non-recirculatory or one-pass perfusion approach has been used. The performance of the apparatus was evaluated with use of three model compounds: antipyrine, lidocaine and ethanol. In addition, oxygen extraction was determined. The steady state extraction ratio (ER) was determined for each compound (and oxygen) as a function of perfusate flow rate (15-35 ml/min) during sequential 45 min perfusion periods. Perfusion experiments lasted for up to 3 hr. The ERs (at 15 ml/min) of ethanol (0.65 +/- 0.15), lidocaine (0.91 +/- 0.01) and oxygen (0.65 +/- 0.10) were dependent upon perfusate flow; whereas, antipyrine ER (0.07 +/- 0.01) was independent of flow. The corresponding values for unbound intrinsic clearances (CLu,int) for antipyrine, ethanol, lidocaine and oxygen were: 1.6, 31.0, 158.0 and 27.5 ml/min, respectively. These findings are consistent with the known hepatic ER values for those compounds reported in the literature. PMID- 10809178 TI - Transport of alovudine (3'-fluorothymidine) into the brain and the cerebrospinal fluid of the rat, studied by microdialysis. AB - Extracellular unbound concentrations of alovudine were sampled by microdialysis in order to study the transport of alovudine between the blood and the brain and the cerebrospinal fluid (CSF) in the rat. The AUC (area under the curve) ratio CSF/blood was higher than the brain/blood ratio after i.v. infusion of alovudine 25mg/kg/hr after a loading dose of 25 mg/kg in 5 minutes (n=4). Neither i.v. infusion of thymidine (25 mg/kg/hr, n=5; 100 mg/kg/hr, n=2) nor acetazolamide (50 mg/kg i.p. bolus followed by 25 mg/kg i.p. every second hour, n=3) influenced the brain/blood AUC ratio after alovudine 25 mg/kg s.c. injection compared to controls (n=5). Finally, perfusion through the microdialysis probe with thymidine (1000 microM, n=3) had also no effect on the brain/blood AUC ratio after alovudine 25 mg/kg s.c. Because neither thymidine nor acetazolamide has significant influence on the ability of alovudine to penetrate the blood-brain barrier in the rat, neither thymidine transport nor carboanhydrase dependent CSF production appear to be major determinants of the blood-brain concentration gradient. Thus, it is concluded that alovudine reaches the extracellular fluid of the brain not by cerebrospinal fluid, but via the cerebral capillaries and that the existence of a concentration gradient over both blood-brain and CSF-brain barrier can probably be explained by the presence of an active process pumping alovudine out from the brain. PMID- 10809179 TI - Platelets induce human umbilical vein endothelial cell proliferation through P selectin. AB - We studied whether platelets could participate in the endothelial cell monolayer regeneration in the case of a vessel damage. Incorporation of [3H]-thymidine into the DNA of human umbilical vein endothelial cells (HUVECs) was measured after 48 h of co-incubation with platelets. The effect of platelets was compared to that of platelet-free supernatants from thrombin-activated platelets that had secreted their active granule constituents. Platelets dose-dependently induced HUVEC proliferation. Platelets preactivated by thrombin induced similar proliferation as did unactivated platelets (proliferation factor = 7 - 8), indicating that preactivation of platelets was not required. Platelets fixed with paraformaldehyde had no effect, suggesting that the platelet mitogenic effect required a mobile, alive membrane. Ketanserine and suramin reduced by at most 30 % the platelet-induced proliferation; supernatants of thrombin-activated platelets caused only minor proliferation (proliferation factor = 2), suggesting that secreted 5-hydroxytryptamine and growth factors poorly contributed to the proliferative effect. When the co-incubation was performed in the presence of an anti P-selectin antibody, the platelet-induced HUVEC proliferation was inhibited. The results suggest that platelet adhesion participate in the control of the endothelial regeneration and that platelet P-selectin is a molecular determinant of the proliferative signal. PMID- 10809180 TI - Social isolation stress enhanced liver metastasis of murine colon 26-L5 carcinoma cells by suppressing immune responses in mice. AB - We investigated the effect of social isolation stress on the formation of experimental liver metastasis resulted from intraportal vein (i.p.v.) injection of colon 26-L5 carcinoma cells in male Balb/c mice, and elucidated some of the underlying mechanism involving the effects of this stress on cellular immunity. Increases in the colony number and tumor burden were observed in the mice socially isolated before and/or after tumor cell challenge, as compared with the group-housed mice. In addition, exposure of mice to 2 weeks of preisolation resulted in decreases in the thymus weight and number of thymocytes by 35.8% and 40.2%, respectively, in comparison with the controls. Reduced proliferative response of splenocytes to various stimuli and suppressed splenic NK activity, as well as decreased macrophage-mediated cytotoxicity, were also found in the mice exposed to social isolation. Thus, these results suggest that social isolation stress enhances tumor metastasis in part via its suppressive effect on the immune system of the host. PMID- 10809181 TI - Actions of angiotensin peptides on the rabbit pulmonary artery. AB - The presence of the angiotensin AT1A-like receptor subtype in the pulmonary artery and AT1B-like receptor subtype in the pulmonary trunk of the rabbit has been reported in two earlier studies. The present study further investigated these receptor subtypes using five other angiotensins (namely angiotensin II, angiotensin III, angiotensin IV, angiotensin-(1-7) and angiotensin-(4-8)). The direct action of the angiotensins on the rabbit pulmonary arterial and trunk sections and the ability of each angiotensin to further contract or relax preconstricted sections of the pulmonary artery and trunk were studied using the organ bath set-up. The effects of angiotensin III on the 3H overflow from re uptaken [3H]noradrenaline in the electrically-contracted rabbit pulmonary arterial and trunk sections were also studied. The contractile response of the arterial and trunk section had the following rank order potency: angiotensin II > angiotensin III > angiotensin IV. The contractile response to these angiotensins was greatly reduced or absent in the pulmonary trunk. Angiotensin II further contracted the preconstricted arterial and trunk sections. In contrast, angiotensin III further contracted the preconstricted arterial section but relaxed the preconstricted trunk section. Angiotensin IV similarly relaxed the preconstricted trunk section but had minimum effect on the preconstricted arterial section. Angiotensin-(1-7) and angiotensin-(4-8) had no effect on both sections. The actions of the three angiotensins were inhibited by losartan, an AT1-selective antagonist. Indomethacin, a cyclo-oxygenase inhibitor, inhibited the relaxation caused by angiotensin III and angiotensin IV in the trunk section. The effects of angiotensin III on the electrically preconstricted sections of the pulmonary trunk and artery were not accompanied by any significant changes in 3H overflow. The differential responses produced by angiotensin II and its immediate metabolites via two positionally located and functionally opposing receptor subtypes suggest that the pulmonary trunk and artery is not a passive conduit but an important regulator of blood flow from the heart to the lung. PMID- 10809182 TI - Extracellular ATP inhibits starvation-induced apoptosis via P2X2 receptors in differentiated rat pheochromocytoma PC12 cells. AB - Apoptosis in neuronal tissue is an efficient mechanism which contributes to both normal cell development and pathological cell death. The present study explored the effects of extracellular ATP on starvation-induced apoptosis in rat pheochromocytoma PC12 cells. Incubation of differentiated PC12 cells with ATP for 6h suppressed apoptosis. 2-Methylthio-ATP, a P2 purinoceptor agonist, was as potent as ATP in suppressing apoptosis, whereas adenosine, ADP, alpha,betamethylene-ATP or UTP was totally ineffective. The suppressive action of ATP was dependent upon the presence of extracellular Ca2+ and blocked by co incubation with the P2 antagonist, suramin. DNA ladder formation, a typical symptom of apoptosis in starved cells, was inhibited by ATP, 2-methylthio-ATP but not by UTP. These results suggest that the inhibitory action of extracellular ATP on apoptotic cell death is mediated via the activation of P2X2 receptors in differentiated PC12 cells. PMID- 10809183 TI - The functions of circulatory polymorphonuclear leukocytes in diabetic patients with and without diabetic triopathy. AB - We determined circulatory polymorphonuclear leukocytes (PMN) functions of superoxide anion production, adhesion and aggregation in 38 type 2 diabetic patients with and without diabetic triopathy. Tumor necrosis factor (TNF)-alpha stimulated superoxide production and N-formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated aggregation in diabetic patients with triopathy were significantly greater than those in diabetics without triopathy. The more diabetic complications existed, the more TNF-alpha-stimulated superoxide was produced by PMN. These results suggest that the activated PMN contributes to a progression of diabetic triopathy in type 2 diabetic patients. PMID- 10809184 TI - Signal transduction efficacy of the highly potent mu opioid agonist 14 methoxymetopon. AB - In search of a truly high-efficacy (i.e., tau > 100) mu opioid analgesic, we determined the efficacy (tau) and apparent in vivo affinity (KA) of the high potency alkoxymorphinan 14-methoxymetopon. However, in the present study, 14 methoxymetopon's efficacy proved to be only 1.5-fold higher than that of morphine (tau, 19 vs. 12). KA values were 2,900 nmol/kg for 14-methoxymetopon and 46,000 nmol/kg for morphine (Ki for [3H]DAMGO binding, 0.33 vs 3.4 nmol/l). Thus, the 24 fold higher potency of methoxymetopon could be fully accounted for by its 16-fold higher apparent in vivo affinity and its only 1.5-fold higher efficacy. Furthermore, the 10-fold higher affinity of 14-methoxymetopon for the mu opioid receptor - as previously determined in radioligand binding assays - was confirmed in the present behavioral tests of thermal antinociception. PMID- 10809185 TI - Antioxidants prevent amyloid peptide-induced apoptosis and alteration of calcium homeostasis in cultured cortical neurons. AB - Beta-amyloid ((A)beta) is a peptide of 39-42 amino acids that is the primary component of plaques in Alzheimer's disease (AD). The mechanism by which (A)beta expresses its neurotoxic effects may involve induction of reactive oxygen species (ROS) and elevation of intracellular free calcium levels. Cultured cortical cells were utilized to study the alterations in calcium homeostasis underlying the neurotoxic effect of (A)beta. Serum supplement B27 and vitamin E were effective in preventing neuronal death as assessed by lactate dehydrogenase (LDH) release, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and number of apoptotic nuclei. In addition, (A)beta-induced cytosolic free calcium ([Ca2+]i) was blocked by antioxidants vitamin E and U83836E, but not by N-methyl D-aspartic acid (NMDA) receptor antagonist MK-801, or by voltage-gated calcium channel blocker nimodipine. Taken together, the results suggest that NMDA receptor and voltage-gated calcium channels are not involved in (A)beta-induced [Ca2+]i increase. This increase appeared to be the result of extracellular calcium influx by some unknown mechanisms. In addition, antioxidants such as B27 were effective in protecting cultured cortical neurons against (A)beta, and correlated with (A)beta attenuation of early calcium response. PMID- 10809186 TI - Sleep and health: research and clinical perspectives. AB - Overall, the Workshop covered most of the principal areas which will be the focus of the Worldwide Project on Sleep and Health. Presentations ranged from the basic science of melatonin receptors to the epidemiology of untreated insomnia, and finally, to the education of primary care physicians. It was emphasized that there is a need for more data, and new experimental paradigms are necessary for successful public health initiatives dealing with sleep disorders. PMID- 10809187 TI - Insomnia in primary care. PMID- 10809188 TI - Sleep-wake state and memory function. AB - Facility of memory formation and retrieval is affected by sleep-wake state. This paper discusses memory function while individuals are awake, but sleepy, at the transitions in state (i.e., wake to sleep and sleep to wake), and in REM and NREM sleep. The basal state of arousal at the time of memory function seems to be critical and the sleepy state, state transitions, and NREM sleep seem to define a continuum of arousal with respect to memory function. While different memory systems have been described, the effect of sleep-wake state on memory systems appears to be non-specific. Finally there is a shift in neurobiology from the NREM to REM state and memory function also appears to change in REM sleep. PMID- 10809189 TI - Public health and insomnia: economic impact. AB - There seems to be a lack of understanding between practitioners and patients on the topic of insomnia. One adult out of four complains of insomnia; however only one insomniac out of four has ever complained about it to their practitioner during a visit made for another problem and only one out of twenty has come to discuss specifically the problem of his or her insomnia. Only a few patients with insomnia take a treatment for it. This gap between the patient's feeling and the practitioner's answer has to be better understood if we want to know why insomnia seems to be so prevalent and what impact it has on society. One aspect may be a question of definition. Insomnia may be considered an ordinary complaint (after one poor night) or as a chronic disease. Practitioners have poor knowledge about sleep disorders and may be embarrassed about coping with an impairment they never specifically learned to manage. The second aspect concerns the impact of insomnia on daytime alertness and performance. While patients usually complain of an impaired daytime functioning with a feeling of fatigue, sleepiness, and risk of mistakes, many studies of insomniacs do not reveal any increased sleepiness or decrease of performance (measured by objective tests) the day following a poor night. Practitioners may therefore find it difficult to understand the real impact of insomnia on daytime functioning. The third aspect is related to the large co-morbidity between insomnia and psychiatric diseases, especially depression and anxiety. It does make it harder for practitioners to define whether the sleep impairment suffered by their patient is the cause for other symptoms or the consequence of an underlying disease. Thus, it makes it all the more difficult for the practitioner to determine which treatment is the most appropriate. These aspects have to be clarified if one wants to estimate the real impact of insomnia on society. It could be useful to both practitioners and patients to have a better understanding on the relationship between poor sleep and daily lives. PMID- 10809190 TI - Hormonal and pharmacological manipulation of the circadian clock: recent developments and future strategies. AB - The mammalian circadian oscillator, located in the suprachiasmatic nuclei of the anterior hypothalamus, serves as the principal source of rhythmic temporal information for virtually all physiologic processes in the organism, including the alternating expression of sleep and wakefulness. Recent studies, in both animal models and human subjects, have demonstrated the important modulation of sleep and wakefulness mediated by the circadian clock. Independent of other factors, notably prior sleep-wake history, the circadian clock potentiates wakefulness (and alertness) at one phase of the diurnal cycle, while facilitating sleep and its attendant processes at the opposite phase. The adaptive advantage of synchronizing sleep-wake behaviors with the daily changes in the external environment is clear. But in a modern world where the constraints of environmental time are less and less important, the circadian clock still imposes rigid boundaries on the timing of sleep and alert wakefulness that are increasingly perceived as limitations on human performance. This conflict underlies the sleep "disorders" of jet lag and shiftwork sleep disruption, problems that are not really diseases at all, but instead reflect normal function of circadian timing in the context of extraordinary demands on sleep-wake scheduling. Whatever their proper classification, both jet lag and shiftwork insomnia represent important societal problems deserving of public health and medical attention. Barring a worldwide rejection of air-travel, jet lag will continue to afflict tens of thousands of people annually. The effects of jet lag on human performance, while typically transient, can nonetheless be significant, affecting commerce, government, and even the outcome of professional sports contests. More important, only a global regression to an agrarian economy will eliminate the problem of tens of millions of workers in this country who regularly attempt to work at night and sleep during the day. In contrast to jet lag, shiftwork produces chronic sleep disruption lasting for the duration of shiftwork exposure. For while individual differences in the ability to adjust to a nocturnal work schedule clearly exist, recent studies suggest that few if any night workers regularly experience restful and restorative day sleep equivalent to that considered normal at night. This chronic sleep limitation is associated with significant increases in a number of consequent problems including sleepiness-related accidents, social disruption, and psychiatric disturbances. In addition, chronic exposure to shiftwork has now been shown to be an independent risk factor for the development of both cardiovascular and gastrointestinal diseases. While these epidemiological studies have not identified the specific aspect of shiftwork that is associated with increased risk of these disorders, the chronic limitation and disruption is foremost among plausible factors. The most important aspect of human circadian physiology that limits adaptation to the extreme schedules inherent in shiftwork and jet travel is the primacy of light among entraining signals, or zeitgebers. Exposure to sunlight for night shiftworkers, or for jet travelers at their destination, results in maintenance (or resetting) of the clock to environmental time. This response can be prevented or overridden with extraordinary avoidance of sunlight or with provision of artificial light of sufficient duration and intensity to negate the sunlight signal, an approach shown to be effective in the treatment of shiftwork sleep disruption. Practical issues sharply limit the application of artificial lighting to all shiftwork settings, however, and the role for a pharmacological chronobiotic agent capable of accomplishing the same end is potentially very large (Copinschi et al., 1995; Jamieson et al., 1998). For example, the effects of zolpidem vs. placebo on sleep, daytime alertness, and fatigue in travelers who complain of jet lag was co PMID- 10809191 TI - Altered patterns of regional cerebral blood flow in patients with Huntington's disease: a SPECT study during rest and cognitive or motor activation. AB - Previous research using functional transcranial Doppler sonography showed that blood flow velocity in the anterior cerebral artery is significantly less in patients with Huntington's disease (HD) than in healthy volunteers while they are completing mazes. The current research used SPECT to study regional cerebral blood flow (rCBF) in patients with HD during rest and maze testing. METHODS: Seven patients with HD and 9 healthy volunteers were injected twice with 0.96 1.15 GBq 99mTc-labeled hexamethylpropylene amine oxime. During the 10 min after injection, subjects either solved mazes or rested with their eyes open while looking at a modified maze. After SPECT, count density was obtained from 11 brain regions and corrected for decay and injected dose. Two types of data generated from this experiment, including absolute regional counts per pixel in the regions of interest and count density computed as a percentage of activity in the lateral cerebellum, were compared between groups. RESULTS: During rest, the absolute regional count density was greater in the HD brains than in the healthy brains (P < 0.001). Count density was typically between 8% and 13% higher in the HD group than in the healthy group. The single exception was the caudate density, for which the 2 groups had similar values. No significant differences in absolute regional count density were observed between groups during maze testing. When rCBF was calculated as a percentage of cerebellar rCBF, analysis of covariance found decreases in HD caudate density (P < 0.001) and orbital frontal cortex density (P < 0.005) during maze testing. Changes in rCBF in the caudate nucleus predicted gene status (P = 0.0007) and correlated with time to complete the mazes (P < 0.05). CONCLUSION: Patients with HD showed an increase in resting rCBF for all brain regions measured except the caudate nucleus. When rCBF was calculated as a percentage of cerebellar blood flow, rCBF in the striatum and orbital cortex in patients with HD was less during maze testing than during rest. Although the cause of these rCBF changes in HD patients is unclear, nitric oxide synthase, a regulator of vasomotor activity, may be involved. PMID- 10809192 TI - Increased cerebral iron uptake in Wilson's disease: a 52Fe-citrate PET study. AB - Toxicity of abundant copper is the main cause of brain and liver tissue damage in patients with Wilson's disease (WD). However, there is also evidence of a disturbed iron metabolism in this genetically determined disorder. This PET study was undertaken to assess cerebral iron metabolism in WD patients. METHODS: We used 52Fe-citrate, which converts to 52Fe-transferrin in blood plasma, to study basic pharmacokinetic features of the cerebral iron transport in 6 WD patients and in 16 healthy volunteers (control subjects). A 2-tissue-compartment model and multiple time graphic plotting were used to calculate 52Fe-transferrin distribution volumes and transport rates. RESULTS: Net iron uptake (Ki) from plasma into brain tissue was significantly (P < 0.001) higher in WD patients (Ki [mean +/- SEM] = 15.1E-05 +/- 7.13E-05 [1/min]) than in healthy volunteers (Ki = 2.66E-05 +/- 0.351E-05 [1/min]). There was no difference of tracer iron distribution in the cerebral plasma volume between patients and healthy volunteers. Iron uptake values resulting from 2 methods to model PET data of patients and healthy volunteers were highly correlated (P < 0.001). CONCLUSION: An abnormally increased cerebral 52Fe-transferrin uptake was found in WD patients. PMID- 10809193 TI - Combined SPECT and diffusion-weighted MRI as a predictor of infarct growth in acute ischemic stroke. AB - In acute ischemic stroke, the infarcted core is surrounded by a zone of tissue that has decreased perfusion. Some of this tissue may be salvaged by prompt, effective treatment. Diffusion-weighted MRI is sensitive in detecting the infarcted tissue, whereas SPECT also detects the hypoperfused tissue around the infarcted core. We studied the potential of combined diffusion-weighted MRI and SPECT to predict infarct growth and clinical outcome in patients not receiving thrombolytic treatment. METHODS: Sixteen patients with acute stroke were examined consecutively with diffusion-weighted MRI and 99mTc-ethyl cysteinate dimer (99mTc ECD) SPECT within 24 h of the onset of symptoms. Follow-up diffusion-weighted MRI was performed on the second day and after 1 wk. The volumes of infarcted and hypoperfused brain tissue were measured from diffusion-weighted MRI and SPECT, respectively. The volume difference between the hypoperfused and infarcted tissue on the first day was compared with the possible increase in infarct volume during the follow-up. Each patient's neurologic status was assessed with the National Institutes of Health Stroke Scale (NIHSS). RESULTS: The volume of infarcted tissue increased from 48 +/- 54 cm3 (mean +/- SD) on the first day to 88 +/- 93 cm3 on the second day (P = 0.001) and to 110 +/- 121 cm3 at 1 wk (P = 0.001). The volume of hypoperfused tissue on the first day was significantly greater than the infarct volume (102 +/- 135 cm3; P = 0.001). The volume difference between the hypoperfused and infarcted tissue on the first day correlated significantly with the infarct growth between the first day and 1 wk (r = 0.71; P < 0.01). Between the first day and 1 wk, the increase of the infarct volume correlated significantly with the change in the NIHSS (r = 0.54; P < 0.05). CONCLUSION: A large hypoperfusion zone around the infarct core in the acute phase of ischemic stroke predicts the infarct growth during the first week, and this correlates significantly with the change in the neurologic status of the patient. Combined diffusion-weighted MRI and SPECT performed within 24 h after the onset of symptoms can be useful in the evaluation of acute stroke to predict infarct growth. PMID- 10809194 TI - Hypofixation and hyperfixation of 99mTc-hexamethyl propyleneamine oxime in subacute cerebral infarction. AB - The relationship between hypofixation and hyperfixation of 99mTc-hexamethyl propyleneamine oxime (99mTc-HMPAO) remains unclear. The purpose of this study was to compare 99mTc-HMPAO SPECT with regional cerebral blood flow (CBF) imaging using 133Xe inhalation in patients with subacute cerebral infarction and to investigate the behavior of 99mTc-HMPAO in the infarct area using dynamic SPECT. METHODS: 133Xe and consecutive 99mTC-HMPAO SPECT studies, the latter of which consisted of dynamic and static scanning, were performed on 51 patients (22 women, 29 men; age range, 40-83 y; mean age, 61 y) with cortical infarction in the middle cerebral artery territory 13-15 d after stroke onset. One region of interest (ROI) was drawn in the infarct area. The control ROI was mirrored to the contralateral side, and the same set of ROIs was applied to all SPECT studies. Fractional fixation of 99mTc-HMPAO in the infarct area was evaluated relatively as the ratio of the infarct-to-control region in 99mTc-HMPAO static tomograms/the ratio of the infarct-to-control region in CBF images using 133Xe inhalation and was classified as hyperfixation when this value was >1.1 and hypofixation when this value was <0.9. To investigate the behavior of 99mTc-HMPAO in the infarct area, the second (36-72 s after tracer injection) and eighth (252-288 s after tracer injection) of 8 dynamic scans were selected, and the washout rate was calculated using the formula: 1 - (mean count in the eighth scan/mean count in the second scan). RESULTS: The infarct area showed hyperfixation of 99mTc-HMPAO when CBF in the area was 35 mL/100 g/min or less and showed hypofixation when CBF was >45 mL/U100 g/min. The washout rate was usually negative when CBF imaging using 133Xe inhalation was <20 mL/100 g/min but was positive when it was >45 mUL/100 g/min. The washout rate was negative when the infarct area showed hyperfixation of 99mTc-HMPAO but was positive when it showed hypofixation. CONCLUSION: 99mTc-HMPAO SPECT underestimates CBF in high-flow regions and overestimates CBF in low-flow regions of subacute cerebral infarction. 99mTc HMPAO hypofixation and hyperfixation are associated with backdiffusion from the brain to blood and gradual accumulation of hydrophilic metabolites, respectively. Dynamic images should be useful for discriminating between 99mTc-HMPAO hypofixation and hyperfixation. PMID- 10809195 TI - Cost-effectiveness analysis of therapy for symptomatic carotid occlusion: PET screening before selective extracranial-to-intracranial bypass versus medical treatment. AB - The St. Louis Carotid Occlusion Study (STLCOS) demonstrated that increased cerebral oxygen extraction fraction (OEF) detected by PET scanning predicted stroke in patients with symptomatic carotid occlusion. Consequently, a trial of extracranial-to-intracranial (EC/IC) arterial bypass for these patients was proposed. The purpose of this study was to examine the cost-effectiveness of using PET in identifying candidates for EC/IC bypass. METHODS: A Markov model was created to estimate the cost-effectiveness of PET screening and treating a cohort of 45 symptomatic patients with carotid occlusion. The primary outcome was incremental cost for PET screening and EC/IC bypass (if OEF was elevated) per incremental quality-adjusted life year (QALY) saved. Rates of stroke and death with surgical and medical treatment were obtained from EC/IC Bypass Trial and STLCOS data. Costs were estimated from the literature. Sensitivity analyses were performed for all assumed variables, including the PET OEF threshold used to select patients for surgery. RESULTS: In the base case, PET screening of the cohort followed by EC/IC bypass on 36 of the 45 patients yielded 23.2 additional QALYs at a cost of $20,000 per QALY, compared with medical therapy alone. A more specific PET threshold, which identified 18 surgical candidates, gained 22.6 QALYs at less cost than medical therapy alone. The results were sensitive to the perioperative stroke rate and the stroke risk reduction conferred by EC/IC bypass surgery. CONCLUSION: If postoperative stroke rates are similar to stroke rates observed in the EC/IC Bypass Trial, EC/IC bypass will be cost-effective in patients with symptomatic carotid occlusion who have increased OEF. A clinical trial of medical therapy versus PET followed by EC/IC bypass (if OEF is elevated) is warranted. PMID- 10809196 TI - Improved detection of individual nodal involvement in squamous cell carcinoma of the esophagus by FDG PET. AB - Because both the number and location of metastatic lymph nodes and the N stage influence survival in esophageal cancer, accurate noninvasive evaluation of individual lymph node groups for the presence of metastasis is essential for therapeutic planning. Therefore, we investigated the accuracy of FDG PET for evaluating individual lymph groups in esophageal cancer patients and compared the results with those of CT and endoscopic sonography (ES). METHODS: Sixty-one consecutive patients with histologically proven primary esophageal carcinoma were studied prospectively with FDG PET. Thirteen patients who were treated nonsurgically were excluded from data analysis. The remaining 48 patients underwent esophagectomy and lymph node dissection. All 48 patients underwent CT scanning, including the lower neck, thorax, and upper abdomen, with intravenous administration of contrast medium. ES was performed in 45 of the patients but was incomplete in 12 patients because of esophageal stenosis. The accuracies of FDG PET, CT, and ES were compared with histologic findings. RESULTS: During surgery, a total of 382 lymph node groups were dissected in 48 patients, of which 100 node groups in 32 patients were malignant on histologic examination. For assessing metastasis to individual groups, FDG PET showed 57% sensitivity, 97% specificity, and 86% accuracy, whereas CT showed 18% sensitivity (P < 0.0001), 99% specificity (P = 0.033), and 78% accuracy (P = 0.003). For N staging, FDG PET was correct in 83% (40/48) of the patients, whereas CT and ES were correct in 60% (29/48; P = 0.006) and 58% (26/45; P = 0.003), respectively. CONCLUSION: FDG PET is more accurate than is CT or ES for evaluating metastasis to individual lymph node groups and for N staging in esophageal cancer and thus may be helpful in determining the therapeutic plan. PMID- 10809197 TI - FDG PET detection of unknown primary tumors. AB - The management of patients presenting with metastases of unknown primary origin remains a clinical challenge despite a large variety of imaging modalities. The aim of this study was to evaluate FDG PET in detecting the sites of primary cancer in these patients. METHODS: Fifty-three patients with metastatic cervical adenopathy (n = 44) or extracervical metastases (n = 9) of unknown primary origin were included after extensive but inconclusive conventional diagnostic work-up. Patients received 370 MBq FDG (10 mCi) intravenously, and whole-body images were acquired at 60 min after injection. Clinical, surgical, and histopathologic findings and complete correlative imaging were used to assess the results. RESULTS: In 27 of 53 patients FDG PET showed focal tracer accumulations corresponding to potential primary tumor sites located in the lungs (n = 12), the palatine tonsil (n = 5), the salivary glands (n = 2), the nasopharynx (n = 1), the oropharynx (n = 3), the maxillary sinus (n = 1), and the larynx (n = 1). Moreover, in 2 patients FDG PET revealed lesions suspected to be tumors in the breast and the ileocolonic area. In 20 (37.8%) of these 53 patients FDG PET was true-positive, identifying the primary tumor in the lungs (n = 10), the head and neck region (n = 8), the breast (n = 1), and the ileocolonic area (n = 1). In 6 of 27 patients FDG PET was false-positive, predominantly identifying suspicious areas in the palatine tonsil (n = 3). One patient denied further diagnostic work up after PET; thus, positive PET could not be evaluated. In 26 of 53 patients PET did not reveal lesions suspected to be the primary. However, primary tumors were not found in these patients at clinical follow-up. CONCLUSION: FDG PET is a valuable diagnostic tool in patients with cancer of unknown primary because it imaged unknown primary tumors in about one third of all patients investigated. In addition, FDG PET assists in both guiding biopsies for histologic evaluation and selecting the appropriate treatment protocols for these patients. PMID- 10809198 TI - Attenuation correction in evaluating renal function in children and adults by a camera-based method. AB - Correction for soft-tissue attenuation is required to evaluate absolute renal function by a camera-based method, and an estimate of renal depth and an attenuation coefficient are commonly used for attenuation correction. The first goal of this study was to develop formulas for the calculation of renal depth in both children and adults. The second goal was to optimize the attenuation coefficient for the estimation of renal accumulation of a 99mTc-labeled agent. METHODS: Renal depth was measured by CT in 74 children and 232 adults and compared with the depth calculated using previously published equations. Multiple stepwise linear regression analysis was conducted using data from children and adults together, and new formulas to calculate renal depth were derived. Using the resulting equations, percentage renal uptake at 2-2.5 min was computed from 99mTc-diethylenetriamine pentaacetic acid (DTPA) renography in 40 children and 92 adults. Percentage renal uptake was assessed using various values of an attenuation coefficient, and an optimized attenuation coefficient was determined to maximize the correlation coefficient between percentage renal uptake and glomerular filtration rate (GFR) measured from 2 blood samples. RESULTS: Although the previously published equations appeared to be acceptable in predicting adult renal depth, they substantially underestimated pediatric renal depth. Renal depth (D, cm) was shown by stepwise regression analysis to depend on the ratio of body weight (W, kg) to body height (H, cm) and was successfully calculated in both children and adults using the derived equations (right: D = 16.778 x W/H + 0.752; left: D = 16.825 x W/H + 0.397). The correlation coefficient between percentage renal uptake of 99mTc-DTPA and measured GFR varied substantially according to the attenuation coefficient used and was the highest (0.947) with an attenuation coefficient of 0.087/cm. CONCLUSION: The equations presented here enabled estimation of renal depth irrespective of the patient's age. Attenuation correction using these equations and the optimized attenuation coefficient appears to aid in evaluating renal accumulation and, consequently, renal function in both children and adults. PMID- 10809199 TI - Effects of preinfarction angina on myocardial injury in patients with acute myocardial infarction: a study with resting 123I-BMIPP and 201T1 myocardial SPECT. AB - METHODS: Recent studies have suggested that patients with preinfarction angina have smaller infarcts and a better in-hospital outcome than those without angina. The mechanisms responsible for limitation of infarct size in the presence of preinfarction angina are unclear. We examined the effects of preinfarction angina on myocardial injury in patients with the first acute myocardial infarction with resting 123I-15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) 201TI myocardial scanning performed within 1 mo of infarction. RESULTS: Of 136 patients tested, 48 (35%) had preinfarction angina within 72 h before infarction, whereas 88 (65%) did not. BMIPP and 201TI defects were scored in 9 segments of the left ventricle (0 = normal, 1 = mild defect, 2 = moderate defect, 3 = severe defect, and 4 = no uptake). The total defect score was defined as the sum of the defect scores. There was no significant difference in percentage diameters of stenoses of infarct-related arteries, collateral circulation, total defect scores for BMIPP, or 201TI between the groups with and without preinfarction angina. However, the ratio of total defect score for 201TI to that for BMIPP was significantly smaller for patients with than for those without preinfarction angina (0.64 +/- 0.21 versus 0.74 +/- 0.25, respectively; P = 0.007). CONCLUSION: Preinfarction angina did not affect the areas at risk in acute myocardial infarction, as shown by BMIPP defect, but decreased necrotic myocardium in the areas at risk, as shown by 201TI defect, and increased metabolically damaged but viable myocardium, as shown by BMIPP and 201TI mismatch through unidentified mechanisms other than collateral circulation (e.g., ischemic preconditioning). PMID- 10809200 TI - Coronary microvascular reactivity to sympathetic stimulation in patients with idiopathic dilated cardiomyopathy. AB - The objective of this study was to assess noninvasively the microvascular reactivity to sympathetic stimulation in patients with idiopathic dilated cardiomyopathy (IDC) and in healthy volunteers, who underwent cardiac catheterization for exclusion of coronary artery disease. METHODS: Myocardial flow was quantified with 13N-ammonia PET and tracer kinetic modeling at rest and in response to cold pressor testing (CPT). Ten healthy volunteers (8 men, 2 women; mean age +/- SD, 50.7 +/- 15 y) and 10 matched patients (8 men, 2 women; mean age, 52.5 +/- 14 y) with IDC (mean left ventricular ejection fraction, 0.30 +/- 0.12) were included in the study. RESULTS: Myocardial perfusion at rest was not significantly different between the groups. However, myocardial vascular resistance (MVR) was significantly lower in IDC patients at rest than in healthy volunteers. In response to CPT a significant decrease in MVR was found in healthy volunteers (1.9 +/- 0.4 to 1.5 +/- 0.4 mm Hg x 100 g/mL; 22% decrease) but not in IDC patients (1.5 +/- 0.4 to 1.4 +/- 0.3 mm Hg x 100 g/mL; 9% decrease). Consequently, the increase of the myocardial blood flow in response to CPT was significantly lower (P < 0.008) in IDC patients (56 +/- 17 to 66 +/- 18 mL/100g/min; 20% increase) compared with healthy volunteers (52 +/- 12 to 80 +/- 30 mL/100 g/min; 52% increase), whereas both showed comparable hemodynamic reactions. CONCLUSION: The data indicate that CPT in combination with 13N PET imaging is a valuable noninvasive tool for assessment of coronary microvascular reaction to sympathetic stimulation in IDC patients. Lower coronary vascular resistance was found in IDC patients at rest compared with healthy volunteers, suggesting possible exhaustion of sympathetically induced dilation of the coronary microvasculature in IDC patients at rest. This mechanism may explain the impaired flow response to cold in IDC patients in the present study. PMID- 10809201 TI - Improvement of cardiac neuronal function after carvedilol treatment in dilated cardiomyopathy: a 123I-MIBG scintigraphic study. AB - Carvedilol can induce important clinical and hemodynamic improvements in patients with chronic heart failure resulting from severe left ventricular (LV) dysfunction. This study examines the impact of carvedilol on cardiac neuronal function using 123I-metaiodobenzylguanidine (MIBG) scintigraphy in dilated cardiomyopathy. METHODS: Twenty-two patients with chronic heart failure (19 men, 3 women; mean age, 54 y; age range, 34-64 y) assessed as New York Hospital Association (NYHA) class II or III and with initial resting radionuclide LV ejection fractions (LVEF) < 0.40 were enrolled in the study. Patients had long histories of symptomatic LV dysfunction despite optimal diuretics and angiotensin converting enzyme inhibitor treatment. Over a 6-mo period, 50 mg/day carvedilol was administered to these patients. Planar 123I-MIBG scintigraphy provided measurements of cardiac neuronal uptake (as heart-to-mediastinum count activity ratio [HMR]), 4h after intravenous injection of 185 MBq MIBG. Hemodynamic, clinical, radionuclide LVEF and HMR data measured at the outset and after 6 mo of carvedilol were compared. RESULTS: Resting heart rate decreased from 81 +/- 13 to 71 +/- 9 bpm (P = 0.003). After carvedilol therapy NYHA functional classification for these patients improved from 2.6 +/- 0.5 to 2.3 +/-0.5 (P = 0.04), LVEF improved from 22% +/- 9% to 30% +/- 13% (P = 0.005), and HMR improved from 145% +/- 23% to 170% +/- 25% (P = 0.0001). CONCLUSION: Carvedilol induces improvements of clinical symptoms and cardiac neuronal and systolic functions in patients with dilated cardiomyopathy and chronic optimal treatment. PMID- 10809202 TI - Limited incremental diagnostic values of attenuation-noncorrected gating and ungated attenuation correction to rest/stress myocardial perfusion SPECT in patients with an intermediate likelihood of coronary artery disease. AB - Either gated myocardial perfusion SPECT or attenuation-corrected SPECT can be used to improve specificity in the diagnosis of coronary artery disease (CAD). We investigated whether attenuation-noncorrected gating and ungated attenuation correction could improve the diagnostic performance of rest/stress perfusion SPECT in patients having an intermediate pretest likelihood of CAD. METHODS: Sixty-eight patients (29 men, 39 women; mean age, 59 +/- 12 y) with coronary artery stenosis > or =70% (1 vessel, n = 13; 2 vessels, n = 18; 3 vessels, n = 8; normal, n = 29) underwent rest attenuation-corrected 201TI SPECT and dipyridamole stress gated attenuation-corrected 99mTc-methoxyisobutyl isonitrile SPECT with an ADAC vertex camera. Three physicians graded the post-test likelihood of CAD for each arterial territory using a 5-point scale (1, normal; 2, possibly normal; 3, equivocal; 4, possibly abnormal; 5, abnormal). The sensitivity, specificity, and areas under receiver-operating-characteristic curves were compared for each operator by 3 methods: attenuation-noncorrected rest/stress SPECT, gated poststress SPECT plus attenuation-noncorrected rest/stress SPECT, and attenuation corrected rest/stress SPECT plus gated poststress SPECT plus attenuation noncorrected rest/stress SPECT. RESULTS: When higher than grade 3 was used as the criterion for CAD, no differences in sensitivity and specificity were found among the 3 methods for each operator. Areas under receiver-operating-characteristic curves for the diagnosis of CAD and stenosis revealed no differences for each modality (P > 0.05 for each comparison). CONCLUSION: In patients with an intermediate risk of CAD, viewing attenuation-noncorrected gated poststress SPECT and ungated attenuation-corrected rest/stress SPECT images did not improve the diagnostic performance for CAD and stenosis. PMID- 10809203 TI - Breast milk excretion of radiopharmaceuticals: mechanisms, findings, and radiation dosimetry. AB - The excretion of radiopharmaceuticals in breast milk is studied to understand excretion mechanisms and to determine recommended breast feeding interruption times for many compounds based on the radiation absorbed dose estimated. A literature review is summarized, providing information on breast milk excretion of many radiopharmaceuticals, including the observed fractions of administered activity excreted and the disappearance half-times. Radiation doses to the infant and to the mother's breasts have been calculated using mathematical models of the activity clearance into milk, with interruption schedules for the nursing infant derived using a dose criteria of 1 mSv effective dose to the infant. In only 9 of the 25 radiopharmaceuticals considered here is interruption in breast feeding thought necessary. However, in the literature, breast milk concentrations of radiopharmaceuticals and half-times varied considerably between subjects, and individual measurements are encouraged to raise confidence in specific cases. The absorbed dose to the mother's breast approaches 10-20 mGy (1-2 rad) for a few nuclides, but most doses are quite low. Therapeutic administration of 131I-NaI is a special case, for which the breast dose for a 5550 MBq (150 mCi) administration could approach 2 Gy (200 rad). In this article, these data are discussed, with the aim of assisting others in evaluating the significance of administration of radiopharmaceuticals to lactating women. An example of a sampling scheme and calculation to determine dose for a specific patient is also developed. PMID- 10809204 TI - Reproducibility and accuracy of gated SPECT for determination of left ventricular volumes and ejection fraction: experimental validation using MRI. AB - Quantitative gated SPECT (QGS) has been used for computation of left ventricular volumes and ejection fraction. This study evaluated, first, the effect of injected dose, time of imaging, and background activity on the reproducibility of QGS and, second, the accuracy of QGS, compared with cine MRI, for determining left ventricular volumes and ejection fractions in dogs with and without perfusion defects. METHODS: Sixteen dogs were subjected to either chronic occlusion of the circumflex artery (group I, no perfusion defect) or acute occlusion of the anterior descending coronary artery (group II, perfusion defect). Both groups underwent serial MRI and SPECT. RESULTS: ( QGS was very reproducible using the automated program (r = 0.99997). Correlation between left ventricular ejection fraction (LVEF) at 15 and 45 min was poor after the low-dose injection (r = 0.54; SE = 9%) and only fair after the high-dose injection (r = 0.77; SE = 5%). Correlation was poor in the presence of significant background activity (r = 0.36; SE = 12%). Correlation between QGS left ventricular volumes and MRI was good for group I (end-diastolic volume, r = 0.86; end-systolic volume, r = 0.81) and only fair for group II (end-diastolic volume, r = 0.66; end systolic volume, r = 0.69). The overall LVEF correlation between QGS and MRI was poor (r = 0.51). QGS LVEF (mean +/- SD, 42% +/- 3%) overestimated MRI LVEF (29% +/- 2%). CONCLUSION: QGS provides a highly reproducible estimate of LVEF. However, QGS is affected by changes in background activity, time of imaging, and injected dose. In the presence of perfusion defects, QGS overestimated volume relative to MRI. The correlation between QGS- and MRI-derived LVEF was poor in this canine model. PMID- 10809205 TI - Noninvasive monitoring of gene transfer using a reporter receptor imaged with a high-affinity peptide radiolabeled with 99mTc or 188Re. AB - Gene therapy protocols require better modalities to monitor the location and level of transferred gene expression. One potential in vivo mechanism to assess gene expression would be to image the binding of a radiolabeled peptide to a reporter receptor that is expressed in targeted tissues. This concept was tested in a tumor model using a replication-incompetent adenoviral vector encoding the human type 2 somatostatin receptor (Ad5-CMVhSSTr2). Expression of the hSSTr2 reporter was imaged using a radiolabeled, somatostatin-avid peptide (P829). METHODS: Bilateral subcutaneous A427 tumor xenografts were established on the flanks of athymic nude mice. These human-origin, non-small cell lung tumors are normally negative for hSSTr2 expression. One tumor was injected directly with Ad5 CMVhSSTr2, whereas the second tumor was injected directly with a control Ad5 vector. The mice were injected intravenously 48 h later with P829 peptide that was radiolabeled to high specific activity with 99mTc (half-life, 6 h) or 188Re (half-life, 17 h). Tumors were frozen and evaluated for somatostatin receptor expression using fluorescein-labeled somatostatin. RESULTS: The accumulation of radiolabeled P829 in hSSTr2-expressing tumors was easily visualized by gamma camera imaging 3 h after injection. Imaging region of interest analyses and biodistribution studies confirmed a 5- to 10-fold greater accumulation of both radiolabeled P829 peptides in the Ad5-CMVhSSTr2-injected tumors versus control tumors injected with control Ad5 vectors. Ad5-CMVhSSTr2-injected tumors accumulated 2.5-3.8 percentage injected dose per gram 3 h after injection. Only Ad5-CMVhSSTr2-injected tumors expressed somatostatin receptors, as determined by immunohistochemistry. CONCLUSION: These studies show the feasibility of imaging a 99mTc-labeled peptide's binding to a reporter receptor after in vivo gene transfer to tumor cells. The 188Re-labeled peptide worked equally well for this imaging approach and offers the additional advantage of energetic beta decay with potential therapeutic efficacy. 99mTc and 188Re are generator produced, an advantage for widespread availability and low cost, and both radioisotopes can be imaged with existing, high-resolution modalities. There is great potential for using 99mTc-labeled peptides for imaging gene transfer with the hSSTr2 reporter receptor, especially when the reporter correlates with the expression of therapeutic genes that can be included simultaneously in the gene therapy vector. PMID- 10809206 TI - Scintigraphic evaluation of experimental chronic osteomyelitis. AB - Assessment of disease activity and disease extent in chronic osteomyelitis remains a difficult diagnostic problem. Radiography is not particularly sensitive. Scintigraphic techniques can be more helpful, but the routinely available agents lack specificity (99mTc-methylene diphosphonate [MDP], 67Ga citrate) or are laborious to prepare (111In-leukocytes). We evaluated the performance of 2 new radiopharmaceuticals, 99mTc-polyethyleneglycol (PEG) liposomes and 99mTc-hydrazinonicotinamide (HYNIC)-immunoglobulin G (IgG), in an experimental model of chronic osteomyelitis. METHODS: Chronic osteomyelitis was induced in rabbits by inserting S. aureus into the right reamed and washed femoral canal. The canal was closed with cement. A sham operation was performed on the left femur. Routine radiographs were obtained immediately after surgery and before scintigraphy. Four weeks after surgery, each rabbit was injected with 37 MBq 99mTc-PEG liposomes, 99mTc-HYNIC-IgG, and 99mTc-MDP on 3 consecutive days and imaged up to 4 (MDP) or 22 (liposomes and IgG) h after injection. On day 4, rabbits received either 18 MBq 111In-granulocytes or 67Ga-citrate and were imaged up to 44 h after injection. Uptake in the infected femur was determined by drawing regions of interest. Ratios of infected-to-sham-operated femur were calculated. After the last image, the rabbits were killed, and the left and right femur were scored for microbiologic and histopathologic evidence of osteomyelitis. RESULTS: 99mTc-PEG liposomes and 99mTc-HYNIC-IgG correctly identified all 6 rabbits with osteomyelitis. 11In-granulocytes and 67Ga-citrate gave equivocal results in 1 infected rabbit. 99mTc-MDP missed 1 case of osteomyelitis. The uptake in the affected region did not differ significantly between the agents, although 99mTc-MDP tended to have higher values (MDP, 4.75 +/ 1.23 percentage injected dose per gram [%ID/g]; 67Ga, 2.05 +/- 0.54 %ID/g; granulocytes, 1.56 +/- 0.83 %ID/g; liposomes, 1.75 +/- 0.76 %ID/g, and IgG, 1.96 +/- 0.27 %ID/g). The ratios of infected-to-normal femur were also not significantly different for the respective radiopharmaceuticals. Radiography visualized only severe osteomyelitis. CONCLUSION: In this rabbit model, 99mTc-PEG liposomes and 99mTc-HYNIC-IgG performed at least as well as 111In-granulocytes and 67Ga-citrate in the localization of chronic osteomyelitis. The ease of preparation, the better image quality, and the lower radiation dose suggest that 99mTc-PEG liposomes and 99mTc-HYNIC-IgG might be suitable alternatives for 67Ga citrate and 111In-granulocytes in the scintigraphic evaluation of osteomyelitis. PMID- 10809207 TI - A comparison of EGF and MAb 528 labeled with 111In for imaging human breast cancer. AB - Our objective was to compare 111In-labeled human epidermal growth factor (hEGF), a 53-amino acid peptide with anti-epidermal growth factor receptor (EGFR) monoclonal antibody (MAb) 528 (IgG2a) for imaging EGFR-positive breast cancer. METHODS: hEGF and MAb 528 were derivatized with diethylenetriamine pentaacetic acid (DTPA) and labeled with 111In acetate. Receptor binding assays were conducted in vitro against MDA-MB-468 human breast cancer cells. Biodistribution and tumor imaging studies were conducted after intravenous injection of the radiopharmaceuticals in athymic mice bearing subcutaneous MCF-7, MDA-MB-231, or MDA-MB-468 human breast cancer xenografts or in severe combined immunodeficiency mice implanted with a breast cancer metastasis (JW-97 cells). MCF-7, MDA-MB-231, JW-97, and MDA-MB-468 cells expressed 1.5 x 10(4), 1.3 x 10(5), 2.7 x 10(5), and 1.3 x 106 EGFR/cell, respectively in vitro. RESULTS: 111In-DTPA-hEGF and 111In DTPA-MAb 528 bound with high affinity to MDA-MB-468 cells (Ka of 7.5 x 10(8) and 1.2 x 10(8) L/mol, respectively). 111In-DTPA-hEGF was eliminated rapidly from the blood with < 0.2% injected dose/g (%ID/g) circulating at 72 h after injection, whereas 111In-DTPA-MAb 528 was cleared more slowly (3%ID/g in the blood at 72 h). Maximum localization of 111In-DTPA-hEGF in MDA-MB-468 tumors (2.2 %ID/g) was 10 fold lower than with 111In-DTPA-MAb 528 (21.6 %ID/g). There was high uptake in the liver and kidneys for both radiopharmaceuticals. Tumor-to-blood ratios were greater for 111In-labeled hEGF than for MAb 528 (12:1 versus 6:1), but all other tumor-to-normal tissue ratios were higher for MAb 528. MDA-MB-468 and JW-97 tumors were imaged successfully with both radiopharmaceuticals, but tumors were more easily visualized using 111In-labeled MAb 528. There was no direct quantitative relationship between EGFR expression on breast cancer cell lines in vitro, and tumor uptake of the radiopharmaceuticals in vivo, but control studies showed that tumor uptake was receptor mediated. CONCLUSION: Our results suggest that the tumor uptake in vivo of receptor-binding radiopharmaceuticals is controlled to a greater extent by their elimination rate from the blood than by the level of receptor expression on the cancer cells. Radiolabeled anti-EGFR MAbs would be more effective for tumor imaging in cancer patients than peptide-based radiopharmaceuticals such as hEGF, because they exhibit higher tumor uptake at only moderately lower tumor-to-blood ratios. PMID- 10809208 TI - Improved imaging of infections by avidin-induced clearance of 99mTc-biotin-PEG liposomes. AB - This article describes the preparation and optimization of biotin polyethyleneglycol (PEG) liposomes and their application in experimental infection models to improve the scintigraphic imaging of infection and inflammation. METHODS: Biotin was coupled to PEG distearoylphosphatidylethanolamine (DSPE) and subsequently incorporated in the PEG liposomes. Biotinylated liposomes were radiolabeled with 99mTc hydrazinonicotinamide. In vitro binding studies were performed to find the optimal biotin concentration in the liposomes. In rats the biodistribution of the 99mTc-biotin-PEG liposomes was compared with the biodistribution of normal (nonbiotinylated) 99mTc-PEG liposomes. Furthermore, in vivo studies in rats were performed to study both the effect of the biotin content and the optimal avidin dose for efficient clearance of the liposomes. Liposomes containing 0.5 or 1.0 mol% biotin-PEG-DSPE were compared in rats with a Staphylococcus aureus infection in the left calf muscle. Avidin was injected 4 h after injection of the liposomes. RESULTS: Biotinylation of the liposomes did not affect their in vivo behavior. All biotin-PEG liposome formulations tested showed good in vitro avidin binding with 50% inhibitory concentrations ranging from 36 to 8 micromol/L. With avidin doses higher than 100 microg, both preparations rapidly cleared from the circulation. As a result, abscess-to-blood ratios increased 5-fold. To illustrate the potential of the avidin-induced clearance of radiolabeled PEG liposomes, we also studied the 99mTc-biotin-PEG liposomes in rabbits with a subcutaneous S. aureus abscess. The infection was visualized only after injection of 100 microg avidin. CONCLUSION: This study shows that biotin-coated 99mTc-PEG liposomes in combination with the injection of avidin can lead to improved imaging of infection or inflammation localized especially in regions with high blood-pool activity. PMID- 10809209 TI - Attenuation correction for cardiac dual-head gamma camera coincidence imaging using segmented myocardial perfusion SPECT. AB - The diagnostic accuracy of cardiac FDG imaging obtained with the dual-head coincidence gamma camera (DHC) is impaired by artifacts induced by nonuniform attenuation. This study proposed a new method (registration and segmentation method for attenuation correction [AC-RS]) to correct these attenuations in the chest region without the need for additional hardware or expensive transmission scanning equipment. METHODS: Before DHC imaging, 99mTc-tetrofosmin SPECT was performed using dual-energy acquisition from both the photopeak and Compton scatter windows. The scatter window images of the 99mTc-tetrofosmin were then registered 3-dimensionally with the cardiac DHC images and segmented into anatomic regions to obtain body and lung contours by applying the optimal threshold method on localized histograms. Theoretic attenuation coefficient values were assigned to the corresponding anatomic regions, and the DHC emission images were reconstructed using these attenuation correction factors. The results were quantitatively evaluated by imaging a cardiac phantom filled with a uniform solution and placed in a chest phantom. Eight nondiabetic subjects were also examined using this technique, and the results were compared with those of measured attenuation-corrected PET images. RESULTS: Use of this technique in phantom and clinical studies decreased the degree of artifacts seen in the inferior wall activity and corrected the emission images. When the results were compared with those of PET scans, the regional relative counts of the uncorrected DHC scan did not correlate with the results of the PET scan. However, the regional relative counts of the AC-RS-corrected DHC scan exhibited a linear correlation with the results of the PET scan (r = 0.73; P < 0.001). CONCLUSION: Reasonably accurate attenuation-corrected cardiac DHC images can be obtained using AC-RS without the need for transmission scanning. PMID- 10809210 TI - Enhancement of the signal-to-noise ratio in H2(15)O bolus PET activation images: a combined cold-bolus, switched protocol. AB - To increase the signal-to-noise ratio (S/N) of H2(15)O bolus PET activation images, we designed and tested a data acquisition protocol that alters the relative distribution of tracer in the uptake and washout phases of the input function. This protocol enhances the S/N gains obtained with conventional switched protocols by combining task switching and the use of a large bolus of blood free of tracer (cold bolus). The cold bolus is formed by sequestering blood in the lower limbs with a double cuff before tracer injection. METHODS: The effect of a combined cold-bolus, switched protocol on the signal from activation images was first simulated using a compartmental model of the uptake of H2(15)O into the brain. Then, the effectiveness of the protocol was investigated in 4 healthy volunteers performing a language task. Each volunteer underwent scanning 12 times: 3 activation/ baseline and 3 baseline/activation scans using the conventional switched protocol and 3 activation/baseline and 3 baseline/activation scans using the combined cold-bolus, switched protocol. The S/N changes introduced when using the cold bolus were analyzed by comparing, across protocols, the magnitude and statistical significance of the activation foci associated with the execution of the language task identified in the averaged subtracted images, and by comparing image noise levels. RESULTS: In the simulated datasets, the combined protocol yielded a substantial increase in the activation signals for scan durations greater than 60 s, in comparison with equivalent signals yielded by the switched protocol alone. In the PET experiments, activation foci obtained using the combined protocol had significantly higher t statistic values than did equivalent foci detected using the conventional switched protocol (mean improvement, 36%). Analysis of the S/N in the averaged subtracted images revealed that the improvements in statistical significance of the activation foci were caused by increases in the signal magnitudes and not by decreases in overall image noise. CONCLUSION: We designed a data acquisition protocol for H2(15)O bolus PET activation studies that combines the use of a tracer-free bolus with a switched protocol. Simulated and experimental data suggest that this combined protocol enhances the S/N gains obtained with a conventional switched protocol. Implementation of the combined protocol in H2(15)O bolus activation studies was easy. PMID- 10809211 TI - Biodistribution and radiation dosimetry of stabilized 99mTc-exametazine-labeled leukocytes in normal subjects. AB - Labeling leukocytes with 99mTc-exametazime is a validated technique for imaging infection and inflammation. A new radiolabeling technique has recently been described that enables leukocyte labeling with a more stable form of 99mTc exametazime. A normal value study of stabilized 99mTc-exametazime-labeled leukocytes has been performed, including biodistribution and dosimetry estimates in normal subjects. METHODS: Ten volunteers were injected with stabilized 99mTc exametazime-labeled autologous leukocytes to study labeled leukocyte kinetics and dosimetry in normal subjects. Serial whole-body imaging and blood sampling were performed up to 24 h after injection. Cell-labeling efficiency and in vivo viability, organ dosimetry, and clearance calculations were obtained from the blood samples and imaging data as well as urine and stool collection up to 36 h after injection. RESULTS: Cell-labeling efficiency of 87.5% +/- 5.1% was achieved, which is similar to or better than that reported with the standard preparation of 99mTc-exametazime. In vivo stability of the radiolabeled leukocytes was also similar to in vitro results with stabilized 99mTc-exametazime and better than previously reported in vivo stability for nonstabilized 99mTc exametazime-labeled leukocytes. Organ dosimetry and radiation absorbed doses were similar with a whole-body absorbed dose of 1.3 x 10(-3) mGy/ MBq. Urinary and fecal excretion of activity was minimal, and visual assessment of the images showed little renal parenchymal activity and no bowel activity up to 2 h after injection. CONCLUSION: Cell labeling and in vivo stability appear improved compared with the leukocytes labeled with the nonstabilized preparation of 99mTc exametazime. There are advantages in more cost-effective preparation of the stabilized 99mTc-exametazime and an extended window for clinical usage, with good visualization of abdominal structures on early images. No significant increase in specific organ and whole-body dosimetry estimates was noted compared with previous estimates using nonstabilized 99mTc-exametazime-labeled leukocytes. PMID- 10809212 TI - Marrow toxicity of 33P-versus 32P-orthophosphate: implications for therapy of bone pain and bone metastases. AB - Several bone-seeking radiopharmaceuticals, such as 32P-orthophosphate, 89Sr chloride, 186Re-1,1 hydroxyethylidene diphosphonate (HEDP), and 153Sm-ethylene diamine tetramethylene phosphonic acid (EDTMP), have been used to treat bone pain. The major limiting factor with this modality is bone marrow toxicity, which arises from the penetrating nature of the high-energy beta particles emitted by the radionuclides. It has been hypothesized that marrow toxicity can be reduced while maintaining therapeutic efficacy by using radionuclides that emit short range beta particles or conversion electrons. In view of the significant clinical experience with 32P-orthophosphate, and the similarity in pain relief afforded by 32P-orthophosphate and 89Sr-chloride, this hypothesis is examined in this study using 32P- and 33P-orthophosphate in a mouse femur model. METHODS: Survival of granulocyte macrophage colony-forming cells (GM-CFCs) in femoral marrow was used as a biologic dosimeter for bone marrow. 32P- and 33P-orthophosphate were administered intravenously, and GM-CFC survival was determined as a function of time after injection and, at the nadir, as a function of injected activity. The kinetics of radioactivity in the marrow, muscle, and femoral bone were also determined. The biologic dosimeter was calibrated by assessing GM-CFC survival at its nadir after chronic irradiation of Swiss Webster mice with exponentially decreasing dose rates of gamma rays (relative biologic effectiveness equivalent to that of beta particles) from a low-dose rate 137Cs irradiator. Dose-rate decrease half-times (Td) (time required for 137Cs gamma ray dose rate to decrease by one half) of 62, 255, and 425 h and infinity were used to simulate the dose rate patterns delivered by the radiopharmaceuticals as dictated by their effective clearance half-times from the mouse femurs. These data were used to experimentally determine the mean absorbed dose to the femoral marrow per unit injected activity. Finally, a theoretical dosimetry model of the mouse femur was developed, and the absorbed doses to the femoral marrow, bone, and endosteum were calculated using the EGS4 Monte Carlo code. RESULTS: When the animals were irradiated with exponentially decreasing dose rates of 137Cs gamma rays, initial dose rates required to achieve 37% survival were 1.9, 0.98, 0.88, and 0.79 cGy/h for dose rate decrease half-times of 62, 255, and 425 h and infinity, respectively. The D37 values were 144 +/- 15, 132 +/- 12, 129 +/- 3, and 133 +/- 10 cGy, respectively, compared with a value of 103 cGy for acute irradiation. When 32P and 33P were administered, the injected activities required to achieve 37% survival were 313 and 2,820 kBq, respectively. Theoretical dosimetry calculations show that 33P offers a 3- to 6-fold therapeutic advantage over 32P, depending on the source and target regions assumed. CONCLUSION: The low-energy beta-particle emitter 33P appears to offer a substantial dosimetric advantage over energetic beta-particle emitters (e.g., 32p, 89Sr, 186Re) for irradiating bone and minimizing marrow toxicity. This suggests that low-energy beta or conversion electron emitters may offer a substantial advantage for alleviation of bone pain as well as for specifically irradiating metastatic disease in bone. PMID- 10809213 TI - Radiation dosimetry for 90Y-2IT-BAD-Lym-1 extrapolated from pharmacokinetics using 111In-2IT-BAD-Lym-1 in patients with non-Hodgkin's lymphoma. AB - Several monoclonal antibodies, including Lym-1, have proven effective for treatment of hematologic malignancies. Lym-1, which preferentially targets malignant lymphocytes, has induced therapeutic responses and prolonged survival in patients with non-Hodgkin's lymphoma (NHL) when labeled with 131. Because radiometal-labeled monoclonal antibodies provide higher tumor radiation doses than corresponding 131I-labeled monoclonal antibodies, the radiation dosimetry of 90Y-2-iminothiolane-2-[p-(bromoacetamido)benzyl]-1,4,7,10-tetraazacyc lododecane N,N',N",N"'-tetraacetic acid-Lym-1 (90Y-21T-BAD-Lym-1) is of importance because of its potential for radioimmunotherapy. Although 90Y has attractive properties for therapy, its secondary bremsstrahlung is less suitable for imaging and pharmacokinetic studies in patients. Thus, the pharmacokinetic data obtained for 111In-21T-BAD-Lym-1 in patients with NHL were used to calculate dosimetry for 90SY-21T-BAD-Lym-1. METHODS: Thirteen patients with advanced-stage NHLwere given a preload dose of unmodified Lym-1 followed by an imaging dose of 111In-21T-BAD Lym-1. Sequential imaging and blood and urine samples obtained for up to 10 d after infusion were used to assess pharmacokinetics. Using 111In pharmacokinetic data and 90Y physical constants, radiation dosimetry for 90Y-21T-BAD-Lym-1 was determined. RESULTS: The uptake of 111In-21T-BAD-Lym-1 in tumors was greater than uptakes in the lung and kidney but similar to uptakes in the liver and spleen. The biologic half-time in tumors was greater than in lungs. The mean radiation dose to tumors was 6.57 +/- 3.18 Gy/GBq. The mean tumor-to-marrow (from blood) radiation ratio was 66:1, tumor-to-total body was 13:1, and tumor-to-liver was 1:1. Images of 111In were of excellent quality; tumors and normal organs were readily identified. Mild and transient Lym-1 toxicity occurred in 3 patients. CONCLUSION: Because of the long residence time of 111In-2IT-BAD-Lym-1 in tumors, high 90Y therapeutic ratios (tumor-to-tissue radiation dose) were achieved for some tissues, but the liver also showed high uptake and retention of the radiometal. PMID- 10809214 TI - Comparing diagnostic accuracy of gamma camera coincidence systems and PET for detection of lung lesions. PMID- 10809215 TI - Does attenuation correction work? PMID- 10809216 TI - Hybrid PET/CT machines: optimized PET machines for the new millennium? PMID- 10809217 TI - Indeterminate adrenal masses. PMID- 10809218 TI - Pediatric renal cell carcinomas: where do they fit in the new histologic classification of renal cell carcinoma? AB - Genetic, immunohistochemical, and histologic data has led to the reclassification of renal cell carcinoma in the last decade. Recent studies suggest that renal cell carcinomas in children and young adults may represent a distinct group of tumors. These tumors have unique genetic findings (most commonly t(x;1)(p11:q21)), a predominantly papillary architecture, numerous calcifications, granular cytoplasm, and a possible relationship with neuroblastoma. PMID- 10809219 TI - Parachordoma exists--but what is it? AB - Parachordoma is a very rare peripheral soft tissue tumor of unknown lineage, which has been described under other names, all of which imply a similarity to chordoma. It forms a circumscribed firm tumor, usually in deep soft tissue, with a variety of histologic patterns and cytologic features, including cords and nests of cells, some of which are vacuolated. The ultrastructure and immunophenotype indicate epithelial differentiation and parachordomas are additionally S-100 protein positive. This tumor is distinct from extraskeletal myxoid chondrosarcoma and probably from soft tissue myoepithelioma. While histologically it somewhat resembles chordoma, parachordoma has a wider range of appearances, and the two neoplasms differ in their detailed cytokeratin immunophenotype and their clinical behavior. Parachordoma is a slowly growing tumor with occasional late recurrence; cases with reported metastasis have not been histologically convincing. This commentary discusses the terminology, origin, and possible nature of this enigmatic neoplasm. PMID- 10809220 TI - Detection of "occult" lymph node metastasis in breast cancer: should pathologists go the extra mile? AB - Approximately 25% of patients with stage I (node negative) breast cancer relapse at a rate similar to those with stage II disease. Inadvertent pathologic "understaging" of lymph node status is one plausible explanation for this phenomenon. While many studies have shown that additional sectioning +/- immunohistochemical staining for epithelial markers increases the rate of detection of small deposits of metastatic carcinoma, few have had sufficient statistical power to examine the impact of "occult" metastasis on clinical outcome. This provocative update by the International Ludwig Breast Cancer Study Group provides support for considering going "the extra mile" to detect lymph node metastasis in postmenopausal patients and those with the lobular subtype of invasive breast cancer. PMID- 10809221 TI - A conceptual shift in the grading of meningiomas. AB - Grading schemes for meningiomas have traditionally designated tumors as "meningioma," "atypical meningioma," or "anaplastic (malignant) meningioma," depending upon the presence of histopathologic features thought to indicate aggressive behavior. In the past, most systems have considered brain invasion by tumor as the best evidence of malignancy. Perry et al. have recently investigated the significance brain invasion as a prognostic feature in meningiomas. The authors studied a series of 116 patients who had been diagnosed previously with "malignant meningioma" due to the presence of brain invasion, histologic anaplasia, or metastasis. On the basis of a multivariate analysis of histopathologic features and their relationship to tumor recurrence and patient survival, the authors concluded that brain invasion should be considered one of the diagnostic features of atypical meningioma. Accordingly, the diagnosis of malignant meningioma should be reserved for those tumors that are frankly anaplastic and/or contain (> = 20 mitoses per 10 high-power fields (HPF). Due in large part to the strength of evidence in this study, the World Health Organization (WHO) has adopted a grading scheme for meningiomas that incorporates many of the authors' proposals. New diagnostic criteria will result in improved reproducibility with fewer diagnoses of malignant meningioma (WHO grade III). PMID- 10809222 TI - Biologic and clinical significance of HER-2/neu (cerbB-2) in breast cancer. AB - HER-2/neu (also known as c-erbB-2) oncogene is a member of the epidermal growth factor receptor family and its amplification is one of the most common genetic alterations associated with human breast cancer. Preclinical studies have suggested that HER-2/neu overexpression enhances metastatic potential of breast cancer cells. Although some discrepancies exist in clinical studies, in general, HER-2/neu amplification is found to be associated with more aggressive clinicopathologic features. HER-2/neu amplification is also associated with drug resistance or sensitivity to specific chemotherapy and hormonal therapy regimens. Advances in breast cancer therapies in recent years have moved towards the development of tumor-specific targeted therapies. Monoclonal antibodies directed against HER-2/neu have been developed and used in clinical practice. These developments necessitate a reliable assay for assessment of HER-2/neu. This article is a review of biologic and clinical significance of HER-2/neu and summarizes HER-2/neu detection methods. PMID- 10809223 TI - Mucosal melanomas: a review. AB - Mucosal melanomas, far fewer in number than melanomas of the skin, manifest a far more aggressive and more rapid life-consuming biologic course. This behavior attends melanomas at any mucosal site, upper aerodigestive tracts, anorectum, and male and female genital tracts. Prognostic factors for both groups of melanoma are similar, but most mucosal melanomas have reached the dangerous limits, e.g., depth of invasion or thickness of melanoma at the time of diagnosis. In general, the mucosal melanomas are also more refractory to therapeutic modalities. In part, this may be due to anatomic restrictions of site and the large size of tumor when first discovered. This review presents a contemporary assessment of melanomas at all of the major mucosal sites. PMID- 10809224 TI - Malignant rhabdoid tumor: A phenotype? An entity?--A controversy revisited. AB - The term malignant rhabdoid tumor (MRT) has been used to describe a heterogeneous group of neoplasms, having in common distinct so-called "rhabdoid" cytologic features. The recent discovery of a candidate tumor suppressor gene for MRT, INI1 on chromosome (Ch)22q11.2, has re-established this neoplasm as a distinct entity. Malignant rhabdoid tumor may arise either de novo from nonneoplastic cells or through tumor progression from other types of neoplasms. These latter tumors, in which other nonrhabdoid tumor components are identified, may be termed composite MRT. In order to avoid misdiagnosing MRT as other types of neoplasia, one must keep in mind three distinct clinicopathologic features--young age of onset, variable histologic and immunohistochemical patterns, and an aggressive infiltrative character. In difficult cases, cytogenetics, fluorescence in situ hybridization (FISH), and molecular genetic analysis may assist in diagnosing MRT. PMID- 10809225 TI - Discovering new hormones, receptors, and signaling mediators in the genomic era. PMID- 10809226 TI - Selection of estrogen receptor beta- and thyroid hormone receptor beta-specific coactivator-mimetic peptides using recombinant peptide libraries. AB - Steroid and thyroid hormone receptors are members of the superfamily of nuclear receptors (NR) that participate in developmental and homeostatic mechanisms by changes in the transcription of specific genes. These activities are governed by the receptors' cognate ligands and through interaction with the components of the transcriptional machinery. A number of coactivator molecules of the steroid receptor coactivator (SRC)/nuclear receptor coactivator (NCoA) family interact with activation functions within NRs through a conserved region containing helical domains of a core LXXLL sequence and, thereby, participate in transcriptional regulation. Using a mammalian-two-hybrid assay, we show that the thyroid hormone receptor beta (TRbeta) and estrogen receptor beta (ERbeta) have different LXXLL motif preferences for interactions with SRC-1. Using large random and focused (centered on the LXXLL motif) recombinant peptide diversity libraries, we have obtained novel peptide sequences that interact specifically with ERbeta or with TRbeta in a ligand-dependent manner. Random sequence libraries yielded LXXLL-containing peptides, and sequence analysis of selected clones revealed that the preferred residues within and around the LXXLL motif vary significantly between these two receptors. We compared the receptor binding of library-selected peptides to that of peptides derived from natural coactivators. The affinities of selected peptides for the ligand binding domains of ERbeta and TRbeta were similar to the best natural LXXLL motifs tested, but showed a higher degree of receptor selectivity. These selected peptides also display receptor-selective dominant inhibitory activities when introduced into mammalian cells. Finally, by directed mutations in specific residues, we were able to alter the receptor binding preference of these peptides. PMID- 10809227 TI - Antiestrogens specifically up-regulate bone morphogenetic protein-4 promoter activity in human osteoblastic cells. AB - Bone morphogenetic protein-4 (BMP-4) plays an important role in the onset of endochondral bone formation in humans, and a reduction in BMP-4 expression has been associated with a variety of bone diseases. Here we describe, by transient transfection assays in bone cells, that the human BMP-4 promoter recently characterized in our laboratory can be stimulated specifically by antiestrogens but not by estrogens or other steroid hormones. This activity is dependent on the presence of the estrogen receptor (ER)-alpha, although the promoter lacks a consensus estrogen-responsive element. No activity was observed in the presence of ERbeta, but synergy was observed when both ER subtypes were cotransfected. The observed stimulation of BMP-4 promoter activity by antiestrogens appeared bone cell specific and was reversed upon addition of estrogens. Since antiestrogens are known to be effective in hormone replacement therapies for postmenopausal women, this observation may help to develop new strategies for treatment and prevention of osteoporosis. PMID- 10809228 TI - An estrogen receptor-alpha splicing variant mediates both positive and negative effects on gene transcription. AB - Analysis of mRNA prepared from a variety of estrogen-responsive cell lines, breast tumor specimens, and normal breast tissue have established that estrogen receptor-alpha (ER alpha) mRNA is typically expressed as a mixture of transcripts. Using PCR amplification, this heterogeneity has been shown to result largely from an imprecise pattern of mRNA splicing that gives rise to a family of correctly processed and exon-skipped ER alpha transcripts. We have reconstructed ER alpha cDNAs representing the single exon-skipped variants ERdeltaE2 through ERdeltaE7 to enable their functional characterization in a well defined cell transfection system. All six of the ER alpha splicing variants support the efficient expression of stable proteins in Cos7 cells, and each shows a characteristic pattern of subcellular distribution. Each of the variants displays a dramatic reduction in DNA-binding activity with a consensus estrogen response element (ERE) in an in vitro gel mobility shift assay. While this DNA-binding defect appears to be complete for ERdeltaE2, ERdeltaE3, ERdeltaE4, and ERdeltaE6, weak DNA binding is observed for ERdeltaE5 and ERdeltaE7. Scatchard analysis of hormone binding demonstrates that among the variants, only ERdeltaE3 binds 17beta estradiol (E2) and does so with an affinity similar to wild-type ER alpha (wt ER alpha). Individual variants cotransfected with the pERE-TK-CAT reporter plasmid [a consensus ERE-driven chloramphenicol acetyltransferase (CAT) reporter gene that is highly responsive to E2-liganded wt ER alpha] were ineffective at inducing CAT expression in ER-negative HeLa cells. Only ERdeltaE5 showed indications of positive transcriptional activity on the pERE-TK-CAT reporter, but this activity was limited to approximately 5% of the activity of wt ER alpha. When variants were expressed simultaneously with wt ER alpha, ERdeltaE3 and ERdeltaE5 were observed to have a dominant negative effect on wt ER alpha transcriptional activity. Like the wild-type receptor, both ERdeltaE3 and ERdeltaE5 interact with steroid receptor coactivator-1e (SRC-1e) in vitro; however, only ERdeltaE3 retained the ability to dimerize with wt ER alpha. Transcription from a region of the ovalbumin promoter, which contains an ERE half site and an AP-1 motif, is positively regulated by liganded wt ER alpha and ERdeltaE3 in phorbol ester-treated, transiently transfected HeLa cells. In both cases, this activity was enhanced by cotransfected cJun. These observations suggest that selected ER alpha splicing variants are likely to exert important transcriptional effects, especially on genes that are regulated by nonconsensus EREs and subject to complex hormonal control. PMID- 10809229 TI - Growth hormone prevents apoptosis through activation of nuclear factor-kappaB in interleukin-3-dependent Ba/F3 cell line. AB - The pro-B Ba/F3 cell line requires interleukin-3 and serum for growth, and their removal results in cell apoptosis. Ba/F3 cells transfected with the GH receptor (GHR) cDNA become able to proliferate in response to GH. To investigate the role of GH in the control of apoptosis, Ba/F3 cells expressing either the wild-type rat GHR (Ba/F3 GHR) or a mutated rat GHR (Ba/F3 ILV/T) were used. We show that Ba/F3 GHR cells, but not parental Ba/F3 or Ba/F3 ILV/T cells, were able to survive in the absence of growth factor. Furthermore, an autocrine/paracrine mode of GH action was suggested by the demonstration that Ba/F3 cells produce GH, and that addition of GH antagonists (B2036 and G120K) promotes apoptosis of Ba/F3 GHR cells. Consistent with survival, the levels of both antiapoptotic proteins Bcl-2 and Bag-1 were maintained in Ba/F3 GHR cells, but not in parental Ba/F3 cells upon growth factor deprivation. Constitutive activation of the transcription factor nuclear factor-kappaB (NF-kappaB), which has been shown to promote cell survival, was sustained in Ba/F3 GHR cells, whereas no NF-kappaB activation was detected in parental Ba/F3 cells in the absence of growth factor. Furthermore, addition of GH induced NF-kappaB DNA binding activity in Ba/F3 GHR cells. Overexpression of the mutated IkappaB alpha (A32/36) protein, known to inhibit NF kappaB activity, resulted in death of growth factor-deprived Ba/F3 GHR cells, and addition of GH was no longer able to rescue these cells from apoptosis. Together, our results provide evidence for a new GH-mediated pathway that initiates a survival signal through activation of the transcription factor NF-kappaB and sustained levels of the antiapoptotic proteins Bcl-2 and Bag-1. PMID- 10809230 TI - Involvement of JAK/STAT (Janus kinase/signal transducer and activator of transcription) in the thyrotropin signaling pathway. AB - TSH is an important physiological regulator of growth and function in thyroid gland. The mechanism of action of TSH depends on interaction with its receptor coupled to heterotrimeric G proteins. We show here that TSH induces the phosphorylation of tyrosine in the intracellular kinases Janus kinase 1 (JAK1) and -2 (JAK2) in rat thyroid cells and in Chinese hamster ovary (CHO) cells transfected with human TSH receptor (TSHR). The JAK family substrates STAT3 (signal transducers and activators of transcription) are rapidly tyrosine phosphorylated in response to TSH. We also find that JAK1, JAK2, and STAT3 coprecipitate with the TSHR, indicating that the TSHR may be able to signal through the intracellular phosphorylation pathway used by the JAK-STAT cascade. TSH increases STAT3-mediated promoter activity and also induces endogenous SOCS-1 (suppressor of cytokine signaling-1) gene expression, a known target gene of STAT3. The expression of a dominant negative form of STAT3 completely inhibited TSH-mediated SOCS-1 expression. These findings suggest that the TSHR is able to signal through JAK/STAT3 pathways. PMID- 10809231 TI - Identification of Phe313 of the gonadotropin-releasing hormone (GnRH) receptor as a site critical for the binding of nonpeptide GnRH antagonists. AB - The dog GnRH receptor was cloned to facilitate the identification and characterization of selective nonpeptide GnRH antagonists. The dog receptor is 92% identical to the human GnRH receptor. Despite such high conservation, the quinolone-based nonpeptide GnRH antagonists were clearly differentiated by each receptor species. By contrast, peptide antagonist binding and functional activity were not differentiated by the two receptors. The basis of the differences was investigated by preparing chimeric receptors followed by site-directed mutagenesis. Remarkably, a single substitution of Phe313 to Leu313 in the dog receptor explained the major differences in binding affinities and functional activities. The single amino acid replacement of Phe313 of the human receptor with Leu313 resulted in a 160-fold decrease of binding affinity of the nonpeptide antagonist compound 1. Conversely, the replacement of Leu313 of the dog receptor with Phe313 resulted in a 360-fold increase of affinity for this compound. These results show that Phe313 of the GnRH receptor is critical for the binding of this structural class of GnRH antagonists and that the dog receptor can be "humanized" by substituting Leu for Phe. This study provides the first identification of a critical residue in the binding pocket occupied by nonpeptide GnRH antagonists and reinforces cautious extrapolation of ligand activity across highly conserved receptors. PMID- 10809233 TI - Structure-function analysis of the Rev-erbA and RVR ligand-binding domains reveals a large hydrophobic surface that mediates corepressor binding and a ligand cavity occupied by side chains. AB - Rev-erbA/RVR are closely related orphan nuclear receptors (NRs) functioning as dominant transcriptional silencers through an association with the nuclear receptor corepressor N-CoR. In contrast with ligand-regulated NRs, Rev-erbA/RVR lack the ligand-binding domain (LBD) C-terminal activation helix, H12. In the case of retinoid acid receptor and thyroid hormone receptor, ligand binding is thought to reposition H12, causing corepressor dissociation and coactivator recruitment, thus leading to transcriptional activation. Here we present homology models of the Rev-erbA/RVR LBDs, which show that the putative ligand cavity is occupied by side chains, suggesting the absence of endogenous ligands. Modeling also revealed a very hydrophobic surface due to the absence of H12, exposing residues from H3, loop 3-4, H4, and H11. Mutation of specific residues from this surface severely impaired the in vitro and in vivo interaction of the Rev erbA/RVR LBD with the receptor-interacting domain of the corepressors N-CoR or its splice variant RIP13delta1, reinforcing the view of the physical association of N-CoR with a LBD surface encompassing H3-H4 and H11. Furthermore, mutations in the LBD surface significantly reduced the ability of Rev-erbA and RVR to function as repressors of transcription. Interestingly, a hydrophobic surface comprised of H3-H4 and H12 in liganded NRs mediates the interaction with coactivators. Hence, it appears that corepressors and coactivators bind to overlapping surfaces of NR LBDs, the conformational change associated with H12 upon ligand binding resulting in a switch from a corepressor- to a coactivator-binding surface. PMID- 10809232 TI - Bcl-w forms complexes with Bax and Bak, and elevated ratios of Bax/Bcl-w and Bak/Bcl-w correspond to spermatogonial and spermatocyte apoptosis in the testis. AB - Bcl-w, a prosurvival member of the Bcl-2 family, is essential for spermatogenesis. However, the mechanisms by which Bcl-w participates in the regulation of apoptosis in the testis are largely unknown. To explore the potential role of Bcl-w in the regulation of apoptosis in the testis, the expression of Bcl-w mRNA and protein during testicular development and spermatogenesis, the dimerization with the proapoptosis members of the Bcl-2 family, and the responses to hormonal stimulation in vitro and apoptosis-inducing signals in vivo were investigated. Both Bcl-w mRNA and protein were detected in Sertoli cells, spermatogonia, and spermatocytes, as well as in Leydig cells. The steady-state levels of Bcl-w mRNA and protein were much higher in Sertoli cells than in spermatogonia and spermatocytes. In the adult rat testis, both Bcl-w mRNA and protein in Sertoli cells displayed a stage-specific expression pattern. Bcl-w could form complexes with Bax and Bak but not with Bad. Bax and Bak were immunohistochemically localized to the same cell types as Bcl-w, but with higher expression levels in spermatocytes and spermatogonia than in Sertoli cells. FSH could up-regulate Bcl-w mRNA levels in the seminiferous tubules cultured in vitro, whereas no effect was observed when testosterone was applied. Three animal models that display spermatogonial apoptosis induced by blockade of stem cell factor/c-kit interaction by a function-blocking anti-c-kit antibody, spermatocyte apoptosis induced by methoxyacetic acid, and apoptosis of spermatogonia, spermatocytes, and spermatids induced by testosterone withdrawal after ethylene dimethane sulfonate treatment were employed to check the changes of Bcl-w, Bax, and Bak protein levels during apoptosis of specific germ cells. In all three models, the ratios of Bax/Bcl-w and Bak/Bcl-w were significantly elevated. The present study suggests that Bcl-w is an important prosurvival factor of Sertoli cells, spermatogonia, and spermatocytes and participates in the regulation of apoptosis by binding proapoptotic factors Bax and Bak. The ratios of Bax/Bcl-w and Bak/Bcl-w may be decisive for the survival of Sertoli cells, spermatogonia, and spermatocytes. PMID- 10809234 TI - GCN5 and ADA adaptor proteins regulate triiodothyronine/GRIP1 and SRC-1 coactivator-dependent gene activation by the human thyroid hormone receptor. AB - We have used yeast genetics and in vitro protein-protein interaction experiments to explore the possibility that GCN5 (general control nonrepressed protein 5) and several other ADA (alteration/deficiency in activation) adaptor proteins of the multimeric SAGA complex can regulate T3/GRIP1 (glucocorticoid receptor interacting protein 1) and SRC-1 (steroid receptor coactivator-1) coactivator dependent activation of transcription by the human T3 receptor beta1 (hTRbeta1). Here, we show that in vivo activation of a T3/GRIP1 or SRC-1 coactivator dependent T3 hormone response element by hTRbeta1 is dependent upon the presence of yeast GCN5, ADA2, ADA1, or ADA3 adaptor proteins and that the histone acetyltransferase (HAT) domains and bromodomain (BrD) of yGCN5 must be intact for maximal activation of transcription. We also observed that hTRbeta1 can bind directly to yeast or human GCN5 as well as hADA2, and that the hGCN5(387-837) sequence could bind directly to either GRIP1 or SRC-1 coactivator. Importantly, the T3-dependent binding of hTRbeta1 to hGCN5(387-837) could be markedly increased by the presence of GRIP1 or SRC1. Mutagenesis of GRIP1 nuclear receptor (NR) Box II and III LXXLL motifs also substantially decreased both in vivo activation of transcription and in vitro T3-dependent binding of hTRbeta1 to hGCN5. Taken together, these experiments support a multistep model of transcriptional initiation wherein the binding of T3 to hTRbeta1 initiates the recruitment of p160 coactivators and GCN5 to form a trimeric transcriptional complex that activates target genes through interactions with ADA/SAGA adaptor proteins and nucleosomal histones. PMID- 10809235 TI - Alteration of a single amino acid in peroxisome proliferator-activated receptor alpha (PPAR alpha) generates a PPAR delta phenotype. AB - Three pharmacologically important nuclear receptors, the peroxisome proliferator activated receptors (PPARs alpha, gamma, and delta), mediate key transcriptional responses involved in lipid homeostasis. The PPAR alpha and gamma subtypes are well conserved from Xenopus to man, but the beta/delta subtypes display substantial species variations in both structure and ligand activation profiles. Characterization of the avian cognates revealed a close relationship between chick (c) alpha and gamma subtypes to their mammalian counterparts, whereas the third chicken subtype was intermediate to Xenopus (x) beta and mammalian delta, establishing that beta and delta are orthologs. Like xPPAR beta, cPPAR beta responded efficiently to hypolipidemic compounds that fail to activate the human counterpart. This provided the opportunity to address the pharmacological problem as to how drug selectivity is achieved and the more global evolutionary question as to the minimal changes needed to generate a new class of receptor. X-ray crystallography and chimeric analyses combined with site-directed mutagenesis of avian and mammalian cognates revealed that a Met to Val change at residue 417 was sufficient to switch the human and chick phenotype. These results establish that the genetic drive to evolve a novel and functionally selectable receptor can be modulated by a single amino acid change and suggest how nuclear receptors can accommodate natural variation in species physiology. PMID- 10809236 TI - Cross-talk between fatty acid and cholesterol metabolism mediated by liver X receptor-alpha. AB - LXR alpha (liver X receptor, also called RLD-1) is a nuclear receptor, highly expressed in tissues that play a role in lipid homeostasis. In this report we show that fatty acids are positive regulators of LXR alpha gene expression and we investigate the molecular mechanisms underlying this regulation. In cultured rat hepatoma and primary hepatocyte cells, fatty acids and the sulfur-substituted fatty acid analog, tetradecylthioacetic acid, robustly induce LXR alpha (up to 3.5- and 7-fold, respectively) but not LXR beta (also called OR-1) mRNA steady state levels, with unsaturated fatty acids being more effective than saturated fatty acids. RNA stability and nuclear run-on studies demonstrate that changes in the transcription rate of the LXR alpha gene account for the major part of the induction of LXR alpha mRNA levels. A similar induction of protein level was also seen after treatment of primary hepatocytes with the same fatty acids. Consistent with such a transcriptional effect, transient transfection studies with a luciferase reporter gene, driven by 1.5 kb of the 5'-flanking region of the mouse (m)LXR alpha gene, show a peroxisome proliferator-activated receptor-alpha dependent increase in luciferase activity upon treatment with tetradecylthioacetic acid and the synthetic peroxisome proliferator-activated receptor-alpha activator, Wy 14.643, suggesting that the mLXR alpha 5'-flanking region contains the necessary sequence elements for fatty acid responsiveness. In addition, in vivo LXR alpha expression was induced by fatty acids, consistent with the in vitro cell culture data. These observations demonstrate that LXR alpha expression is controlled by fatty acid signaling pathways and suggest an important cross-talk between fatty acid and cholesterol regulation of lipid metabolism. PMID- 10809237 TI - Androgen induction of cyclin-dependent kinase inhibitor p21 gene: role of androgen receptor and transcription factor Sp1 complex. AB - Previous studies have shown that androgen up-regulates expression of the p21 (WAF1, CIP1, SDI1, CAP20) gene, which contains a canonical androgen response element (ARE) in its proximal promoter region. We undertook the current studies to determine whether elements in the p21 promoter other than the ARE mediate androgen action. We found that deletion of the ARE did not completely abolish the promoter responsiveness to androgen, suggesting that additional cis-regulatory elements within the p21 core promoter may also be involved in androgen responsiveness. The p21 core promoter is GC-rich and contains six binding sites for transcription factor Sp1. We determined whether one or more of these Sp1 sites mediate androgen responsiveness of the p21 promoter. To do so, we used a transient transfection assay with p21 promoter-luciferase reporter constructs. The reporter activity of a construct lacking the ARE but containing all six Sp1 sites was induced approximately 3-fold by androgen. Mutation of Sp1-3 nearly eliminated basal promoter activity as well as androgen responsiveness, whereas deletion of Sp1-1 and Sp1-2 sites and mutation of Sp1-4, Sp1-5, and Sp1-6 sites had relatively little effect. We also used the mammalian one-hybrid assay and coimmunoprecipitation assay to show that androgen receptor (AR) and transcription factor Sp1 interact with one another. The current studies suggest a model in which AR and transcription factor Sp1 not only bind to their respective consensus sites within the p21 promoter, but also complex with one another, thereby recruiting coactivators and general transcription factors and inducing p21 transcription. PMID- 10809238 TI - Homocysteine and cardiovascular risk in dialysis patients. PMID- 10809239 TI - Homocysteine, vitamin B12, and serum and erythrocyte folate in peritoneal dialysis and hemodialysis patients. AB - BACKGROUND: Plasma homocysteine (Hcy) is an independent risk factor for cardiovascular disease. High levels of plasma Hcy have been observed in end-stage renal disease patients. Few studies have compared peritoneal dialysis (PD) and hemodialysis (HD) patients and few data are available on erythrocyte folate (ery F) levels in dialysis patients. OBJECTIVES: To evaluate plasma Hcy concentrations, vitamin B12 (B12), and folate status in dialysis patients; to analyze the possible causes of high Hcy levels; to follow up changes in folate and B12 concentrations after 6 months. DESIGN: A cross-sectional observational study. SETTING: Nephrology division and laboratory of hematology in a university and clinical research hospital. PATIENTS: The study included 82 patients treated with PD for 37 + 37 months and 70 patients treated with HD for 136 + 95 months. LABORATORY METHODS: Plasma Hcy was measured by the immunoenzymatic IMx Hcy FPIA method (Abbott Laboratories, Diagnostic Division, Abbott Park, IL, U.S.A.), serum folate (s-F) and ery-F by the Stratus folate fluorometric enzyme-linked assay, and B12 by the Stratus vitamin B12 fluorometric enzyme-linked assay (DADE Behring, Newark, DE, U.S.A.). RESULTS: Ninety-six percent of PD and 97% of HD patients had Hcy levels above the cutoff (13.5 micromol/L). Homocysteine level was higher in HD than in PD patients, while the prevalence of hyperhomocysteinemia was similar with the two techniques. Erythrocyte folate was significantly higher in PD (1333 +/- 519 pmol/L) than in HD (1049 +/-511 pmol/L, p < 0.01). Statistically significant correlations were observed between Hcy and B12, s-F, ery-F, and dialysis duration. Multivariate analysis showed a strong correlation between s-F and Hcy. After 6 months there were no differences in Hcy, B12, s-F, and ery-F levels. CONCLUSIONS: Plasma Hcy levels were high in more than 95% of our dialysis patients, with no relation to the type of dialysis. Vitamin B12 and folate were normal in the majority of our patients. However, serum folate was the major determinant of Hcy levels. Such a relation between Hcy and folate suggests that levels of folate within the reference interval are inadequate for dialysis patients. PMID- 10809240 TI - Peritoneal kinetics and mesothelial markers in CCPD using icodextrin for daytime dwell for two years. AB - OBJECTIVE: To evaluate the safety, efficacy, and biocompatibility of icodextrin (Ico), continuous cycling peritoneal dialysis (CCPD) patients were treated for 2 years with either Ico- or glucose (Glu)-containing dialysis fluid for their daytime dwell (14 - 15 hours). Prior to entry into the study, all patients used standard Glu solutions (Dianeal, Baxter BV, Utrecht,The Netherlands). DESIGN: Open, randomized, prospective two-center study. SETTING: University hospital and teaching hospital. PATIENTS: Both established patients and patients new to CCPD were included. A life expectancy of more than 2 years, a stable clinical condition, and written informed consent were necessary before entry. Patients aged under 18 years or with peritonitis in the previous month, and women of childbearing potential unless taking adequate contraceptive precautions, were excluded. Thirty-eight patients entered the study (19 Glu, 19 Ico). MAIN OUTCOME MEASURES: Daytime dwell peritoneal effluents were collected every 3 months in combination with other study variables (clinical data, laboratory measurements, dialysis-related data, and urine collection). Peritoneal transport studies were carried out every 6 months. RESULTS: In Glu- and Ico-treated patients, peritoneal transport of low molecular weight solutes and protein clearances neither changed during follow-up nor differed between the two groups. Peritoneal membrane markers (CA125, interleukin-8, carboxyterminal propeptide of type I procollagen, and aminoterminal propeptide of type III procollagen) measured in effluents did not differ between the groups and did not change over time. All these markers showed a dialysate/plasma ratio of more than 1, suggesting local production. Residual renal function remained stable during follow-up and adverse clinical effects were not observed. CONCLUSIONS: Peritoneal membrane transport kinetics and markers remained stable in both groups over a 2-year follow-up period. Membrane markers were higher in effluents than in serum, suggesting local production. No clinical side effects were demonstrated. Icodextrin was a well-tolerated effective treatment. PMID- 10809241 TI - The effect of small solute clearances on survival of anuric peritoneal dialysis patients. AB - OBJECTIVE: Primarily, to determine whether peritoneal small solute clearance is related to patient and technique survival among anuric peritoneal dialysis [continuous ambulatory (CAPD) and automated peritoneal dialysis (APD)] patients. A secondary goal was to describe the ability to attain Dialysis Outcomes Quality Initiative (DOQI) targets among anuric patients on peritoneal dialysis. DESIGN: Retrospective cohort study via chart reviews. SETTING: Peritoneal Dialysis Unit of Toronto Hospital (Western Division). PATIENTS: The study included 122 CAPD and APD patients between January 1992 and September 1997, with 24-hour urine volume less than 100 mL, or renal creatinine clearance (CCr) less than 1 mL/minute. Adequacy data were available for 115 patients. OUTCOME MEASURES: Mortality and technique failure (TF). Regression analysis was used to estimate the mortality and TF rate ratios (RR) for peritoneal Kt/V urea (pKt/V) and pCCr, adjusting for age, gender, diabetes, months of follow-up prior to anuria, albumin, transport status, coronary artery disease, cardiovascular disease, and peripheral vascular disease. RESULTS: Fifty seven per cent (51/89) of patients on CAPD and 81% (21/26) on APD had a weekly pKt/V > or = 2 and > or = 2.2, respectively (DOQI targets); whereas only 35% on CAPD (31/89) and 35% (9/26) on APD had a weekly pCCr > or = 60 U1.73 m2 and 66 L/1.73 m2, respectively. Median follow-up times among patients were 16.5 and 19.5 months pre- and postanuria, respectively. Patients with pKt/V > or = 1.85 experienced a strong decrease in patient mortality (RR = 0.54, p= 0.10); the effect was less pronounced for pCCr > or = 50 L/1.73 m2 (RR = 0.63, p = 0.25). No relationship was observed between pKt/V or pCCr and TF. CONCLUSION: Mortality was noticeably less frequent among patients with a pKt/V > or = 1.85 compared with those with a Kt/W < 1.85 (p = 0.10). Given the magnitude of the association, the failure to observe statistical significance relates to the size of the patient cohort. Our results imply that it is, in fact, possible to achieve DOQI targets among anuric patients on peritoneal dialysis. PMID- 10809242 TI - Relationship between fill volume, intraperitoneal pressure, body size, and subjective discomfort perception in CAPD patients. Mexican Nephrology Collaborative Study Group. AB - OBJECTIVE: To determine changes in intraperitoneal pressure (IPP) when dialysate fill volume is increased from 2.0 L to 2.5 L to 3.0 L per exchange, and to evaluate the relationship with subjective discomfort perception. DESIGN: Cross sectional survey. SETTING: Seven Mexican hospital-based dialysis centers. PATIENTS: Eighty-one adult patients on continuous ambulatory peritoneal dialysis (CAPD) without restriction criteria for age, gender, or time on dialysis, were studied. Patients seropositive for HIV or hepatitis B, and those with cancer or receiving immunosuppressive drugs were excluded. Participants were studied as outpatients. MAIN MEASURES: Blindly and in random order, 2.0-, 2.5-, and 3.0-L volumes of dialysate were infused consecutively. Body surface area (BSA) was calculated from patient height and weight. IPP was assessed with the patient lying supine, measuring the height of the dialysate column inside the peritoneal dialysis bag tubing. Blood pressure and subjective discomfort perception (using a visual analog scale of 0-100 mm) were also evaluated and registered after each of the three exchanges. RESULTS: The IPP rose with each increase of dialysate volume and was higher in males than in females for each fill volume level. For males IPP was 18.9 +/- 6.9, 20.8 +/- 7.1, and 22.9 +/- 7.5 cm H2O; and for females it was 16.5 +/- 5.7, 18.4 +/- 5.5, and 19.7 +/- 6.2 cm H2O for 2.0-, 2.5-, and 3.0-L fill volumes respectively (p < 0.01 among fill volumes and between genders). Intraperitoneal pressure showed significant negative correlation with the fill volume corrected for patient body size as reflected by the dialysate volume/ BSA ratio (r= -0.393, p < 0.01; r= 0.319, p < 0.01; and r= -0.274, p < 0.02 for 2.0-, 2.5-, and 3.0-L fill volumes respectively). Discomfort score rose as fill volume rose, with a median of 0, 2.5, and 13.0 for 2.0-, 2.5-, and 3.0-L fill volumes respectively (p< 0.001). It is interesting, however, that with 2.5-L and 3.0-L dialysate infusion volumes, 64% and 44% of the patients, respectively, had no discomfort at all. CONCLUSION: Dialysate volume increase is associated with higher IPP, which is modulated by the gender and body size of the patients. Although the mean discomfort score was higher with larger dialysate volumes, there was no significant correlation between discomfort and IPP or the dialysate volume/BSA ratio. Many patients had no discomfort with 2.5-L or even with 3.0-L dialysate infusions; theoretically, they can be treated with larger volumes. PMID- 10809243 TI - Clinical outcomes of immediate full-volume exchange one year after peritoneal catheter implantation for CAPD. AB - OBJECTIVE: To evaluate actual risk of complications and 1-year catheter survival of immediate full-volume 2000-mL dialysate exchange after peritoneal catheter implantation. DESIGN AND SETTING: A prospective randomized comparative study in a university hospital kidney center. PATIENTS: The study included 59 end-stage renal disease patients entering into a continuous ambulatory peritoneal dialysis program between January 1996 and December 1997. INTERVENTIONS: In group 1 (n = 21), exchange volume was gradually increased from 500 mL per 3 hours to full volume exchange over 13 days. In group 2 (n = 38), full-volume exchange per 6 hours was performed from the day of catheter implantation. Bed rest with minimal activity was recommended for the first 3 days in both groups. MAIN OUTCOME MEASURES: Occurrence of catheter-related complications such as dialysate leakage, drainage failure, malposition, infectious complications, and, ultimately, catheter loss were observed for 1 year. RESULTS: After straight Tenckhoff catheter implantation, pericatheter dialysate leaks occurred in 9.5% of group 1 and in 10.5% of group 2 patients (no significance, NS). Within 1 month of implantation, the incidences of tunnel or exit-site infection, catheter malposition, and outflow failure were not significantly different between the two groups. Abdominal discomfort was noted in 2 patients from group 2. Peritonitis occurred in 9.5% and 5.3% (NS) within 2 weeks of catheter implantation, and in 14.3% and 10.5% (NS) after more than 2 weeks in group 1 and group 2, respectively. Early catheter loss within 1 month occurred in 4.8% of patients in group 1 and 5.3% in group 2 (NS). The causes of loss were persistent leakage in group 1, and persistent leakage combined with tunnel infection and outflow failure in group 2. The duration of hospitalization from the day of catheter insertion to discharge was definitely shorter in group 2 compared to group 1 (9.9 +/-0.6 days vs 15.0 +/- 0.8 days, p < 0.001). For 1-year follow-up, the frequencies of peritonitis were 0.062 +/- 0.015/patient-month in group 1 and 0.076 +/- 0.018/patient-month in group 2 (NS). The actual 1-year catheter survival was 85.7% in group 1 and 84.2% in group 2 (NS). CONCLUSIONS: One-year catheter survival after immediate full-volume peritoneal dialysis exchange was 84.2%. This study did not show any evidence that immediate full-volume exchange causes more short- or long-term complications compared to the stepwise volume increment method. PMID- 10809244 TI - Recurrence of hyperparathyroidism after total parathyroidectomy and autotransplantation in peritoneal dialysis patients. AB - OBJECTIVE: To evaluate the effectiveness of total parathyroidectomy (PTX) with autotransplantation in the treatment of secondary hyperparathyroidism (HPT), and to assess recurrence rate of HPT in this peritoneal dialysis (PD) population. DESIGN: A retrospective study in a single home PD unit. PATIENTS: Between 1994 and 1998, 19 of 574 patients on PD underwent PTX for treatment of secondary HPT. MAIN OUTCOME MEASURES: Clinical and biochemical improvement, recurrence of HPT, improvement in anemia post-PTX. RESULTS: Nineteen (3.3%) patients required PTX between 1994 and 1998. These 5 men and 14 women ranged in age from 22 to 66 years; they had been on maintenance PD pre-PTX for 47.5 +/- 38.1 months, and were followed for 26.1 +/- 15.5 months post-PTX. Sixteen patients had temporary hypocalcemia that was managed by oral (n = 10) or intravenous (n = 6) calcium supplements and calcitriol, while 3 patients had severe "hungry bone" syndrome postoperatively. One patient had recurrent laryngeal nerve palsy post-PTX. Bone pain disappeared in all 12 patients. Pruritus improved in 12/13 patients; fatigue improved in 15/16 patients. Comparison showed significant differences between hemoglobin and hematocrit values 1 month pre-PTX and 12 months post-PTX (p < 0.05). Parathyroid hormone (PTH) level in 15 (79%) patients returned to normal (< or = 7.6 pmol/L) during the first month post-PTX. In 5/12 (42%) patients, PTH level was < or = 7.6 pmol/L 2 years post-PTX, while in 2/12 (17%), PTH was > 22.8 pmol/L (three times normal) 2 years post-PTX, and 3/5 (60%) patients had a PTH > 22.8 pmol/L 3 years post-PTX. CONCLUSIONS: Total PTX with autotransplantation is associated with a tendency for recurrence of HPT. Our findings suggest that total PTX with autotransplantation may be an ineffective procedure in controlling HPT over the long term. PMID- 10809245 TI - Exit-site care with ciprofloxacin otologic solution prevents polyurethane catheter infection in peritoneal dialysis patients. AB - OBJECTIVE: Mupirocin ointment and antiseptics are standard cleansing agents in routine exit-site care of peritoneal dialysis (PD) catheters, but these agents have a deleterious effect on polyurethane devices. We assessed the effectiveness of topical use of ciprofloxacin otologic solution for preventing exit-site infection (ESI) in PD patients with polyurethane catheters. DESIGN: Prospective study. SETTING: Service of Nephrology of an acute-care teaching hospital in Galdacano, Bizkaia, Spain. PATIENTS: A total of 164 patients with polyurethane catheters inserted was studied from start of continuous ambulatory PD to the end of a 24-month period. Patients were divided into two groups according to exit site treatment protocols. INTERVENTION: Patients in group 1 (n = 86) were instructed on daily exit-site care with soap and water only; whereas patients in group 2 (n = 78) cleansed with soap and water, followed by application of a single-dose vial of 0.5 mL ciprofloxacin (1 mg) for application around the insertion site. MAIN OUTCOME MEASURES: Episodes of ESI and peritonitis. RESULTS: There were 67 episodes of ESI among patients in group 1 versus 9 episodes among patients in group 2 (p < 0.05), resulting in a rate of 0.41 and 0.06 episodes per patient-year of exposure, respectively (p < 0.001). Staphylococcus aureus ESI rate was 0.34 in group 1 versus 0.06 in group 2 (p = 0.001). Infections caused by Pseudomonas aeruginosa and other pathogens occurred in 11 patients in group 1 and in no patients in group 2 (p = 0.05). Peritonitis due to S. aureus ESI was significantly less frequent among patients treated with ciprofloxacin (1 vs 9 cases, p = 0.001). Removal of the catheter was necessary in 5 patients in group 1 and in no patients in group 2 (p < 0.05). CONCLUSION: Daily application of ciprofloxacin otologic solution at the exit site of PD patients with polyurethane catheters inserted significantly reduces the rate of ESI caused by S. aureus and other organisms, particularly P. aeruginosa. PMID- 10809246 TI - Outcome following staphylococcal peritonitis. AB - OBJECTIVE: Staphylococcus spp predominate as the causative pathogen of continuous ambulatory peritoneal dialysis (CAPD)-related peritonitis.This study evaluated the difference in morbidity and mortality between peritonitis caused by S. aureus and coagulase-negative staphylococci (CoNS). DESIGN: Prospective observational study. SETTING: A single regional dialysis unit in a teaching hospital. PATIENTS: Thirty-seven patients had S. aureus peritonitis and 65 patients had CoNS peritonitis between July 1990 and November 1995. MAIN OUTCOME MEASURES: Using the first recorded episode of peritonitis, survival analysis was performed for time to (1) death, (2) removal of peritoneal dialysis catheter, and (3) change to hemodialysis. Abdominal complications were recorded for the first and subsequent episodes. RESULTS: No difference in time to death was demonstrated for the two groups (p = 0.79), although two deaths that occurred during therapy for peritonitis were attributable to S. aureus infection. In addition, 5 patients developed serious abdominal complications related to an episode of S. aureus peritonitis. Patients with S. aureus peritonitis had a shorter time to both peritoneal dialysis catheter removal (p = 0.004) and change to hemodialysis (p = 0.014). The change in mode of dialysis was independent of catheter loss. CONCLUSION: This study highlights the serious nature of S. aureus peritonitis and confirms the need for effective preventive measures against infection by this pathogen. PMID- 10809247 TI - The longitudinal effect of a single peritonitis episode on peritoneal membrane transport in CAPD patients. AB - OBJECTIVE: To evaluate the longitudinal effect of a single peritonitis episode on peritoneal membrane transport. DESIGN: A prospective longitudinal study. SETTING: Department of nephrology in a university hospital. PATIENTS: Eighteen continuous ambulatory peritoneal dialysis patients with peritonitis. METHODS: Peritoneal transport for low, middle, and high molecular weight (MW) solutes was evaluated by peritoneal equilibration test (PET). The first PET was performed on the day following the diagnosis of peritonitis. The test was repeated at weeks 1, 2, 4, 12, and 24 and the results were compared to baseline PET data obtained before peritonitis. In addition, dialysate CA125 concentration and leukocyte count were measured. RESULTS: During peritonitis there were significant increases in dialysate-to-plasma (D/P) ratios for all low, middle, and high MW solutes except potassium, and decreases in D4/D0 glucose ratio and ultrafiltration (UF) volume. Over the subsequent 2 weeks, solute transport gradually decreased to the baseline values then remained unchanged during follow-up. Although net UF volume demonstrated a similar course during the study, it did not completely return to the baseline value. No decrease in D/P sodium ratio was found at 60 minutes during the PET performed 24 weeks after peritonitis. The percent change in solute transport during peritonitis compared to baseline value was significantly correlated with a solute's MW (r = 0.776, p = 0.014). The slope of the regression line for D/P ratios versus MW, in double logarithmic scale, before peritonitis ( 0.73 +/- 0.09) was steeper than the slope during peritonitis (-0.59 +/- 0.08). CONCLUSIONS: These findings indicate that a single peritonitis episode does not permanently affect peritoneal solute transport. However, the loss of net UF that accompanies peritonitis is not completely recovered, probably due to impairment of transcellular water transport. The transport changes associated with peritonitis may be due to the combined effect of increased effective peritoneal surface area and intrinsic permeability. Our findings suggest that the latter mechanism seems to be more important. PMID- 10809248 TI - Pharmacokinetics of intraperitoneal piperacillin/tazobactam in patients on peritoneal dialysis with and without pseudomonas peritonitis. AB - OBJECTIVE: The objective of this study was to assess the pharmacokinetics of intraperitoneal (IP) administration of the antibiotic combination piperacillin/tazobactam (PIP/TAZ) to patients on chronic ambulatory peritoneal dialysis (CAPD) with and without pseudomonas peritonitis. DESIGN: Open-labeled study. SETTING: The study was carried out in the CAPD unit of Assaf Harofeh Medical Center, Zerifin, Israel. PATIENTS AND METHODS: Six patients participated in the study, 4 had pseudomonas peritonitis, all were given an IP loading dose of 4 g/0.5 g PIP/TAZ. Twenty-four hours after the initial dose, a maintenance dose of 0.5 g/0.0625 g PIP/TAZ was administered with each dialysate exchange for a period of 1 week. The patients without peritonitis received only the loading dose. High performance liquid chromatography was used to determine the concentrations of PIPITAZ in plasma obtained at 0, 30, 60, 90, 120, 360, 480, 600, 720, and 1440 minutes after administration. Samples of the dialysate fluid for determination of PIP/TAZ concentration were collected at 6,10,14, 24, and 72, 120, and 168 hours. RESULTS: After the loading dose, the highest plasma PIP concentration (Cmax) was 51.6 t 21.25 Lig/mL and appeared at 1.5 = 0.45 hours (t,,a). During the maintenance period plasma PIP concentration was 5.2 t 4.75 Lg/mL. Tazobactam was detected in the plasma of 1 patient only. The concentration of TAZ in the dialysate fluid during the maintenance period was 2.3 t 0.5 ig/mL. CONCLUSIONS: Piperacillin administered IP at 4 g reached plasma concentrations comparable to intravenous administration and considered therapeutic (above the MIC90 for Pseudomonas aeruginosa) in CAPD patients with or without peritonitis. The maintenance dose, however, should be augmented. Tazobactam could not be detected in the plasma of most patients and the therapeutic implications of IP administration of TAZ cannot be directly correlated to intravenous administration. PMID- 10809250 TI - Epidermal down-growth in exit site of peritoneal dialysis catheter and a new peritoneal access technique. PMID- 10809249 TI - Use of intraperitoneal cefepime as monotherapy in treatment of CAPD peritonitis. PMID- 10809251 TI - Leukocyte esterase reagent strips for early detection of peritonitis in patients on peritoneal dialysis. PMID- 10809252 TI - Effects of automated peritoneal dialysis on residual urinary volume. PMID- 10809253 TI - A case of peritonitis due to Rothia dentocariosa in a CAPD patient. PMID- 10809254 TI - "Sick-bag" syndrome. PMID- 10809255 TI - Total lymphocyte count as a prognostic index in CAPD patients. PMID- 10809256 TI - Improvement of diabetes and healing of vascular ulcers after change from peritoneal dialysis to hemodialysis. PMID- 10809258 TI - Literature. PMID- 10809257 TI - The trials and tribulations of an adolescent patient with end-stage renal disease. PMID- 10809259 TI - Reduced platelet activation and thrombosis in extracorporeal circuits coated with nitric oxide release polymers. AB - OBJECTIVE: To determine whether the use of nitric oxide (NO)-releasing polymers coated onto the inner surface of extracorporeal circuits can reduce platelet consumption and activation in the absence of systemic heparinization using a rabbit model of venovenous extracorporeal circulation. DESIGN: Prospective, controlled trial. SETTING: Research laboratory at an academic medical institution. SUBJECTS: New Zealand White Rabbits. INTERVENTIONS: Anesthetized, tracheotomized, and ventilated New Zealand White rabbits were injected with freshly prepared, 111In(oxine)3 labeled single donor platelets through the external jugular vein. After baseline measurements, these animals were placed on venovenous extracorporeal circulation through a 1-m control circuit or NO test circuit for 4 hrs at a blood flow rate of 109-118 mL/min via roller pump. Four groups were studied: systemically heparinized control circuits, systemically heparinized NO test circuits, nonheparinized control circuits, and nonheparinized NO test circuits. Platelet counts, fibrinogen levels, and plasma free indium levels were measured hourly. Circuits were rinsed and retained for gamma counting after the 4-hr run or when the circuit clotted. Four animals, one from each group, did not receive radiolabeled platelets so that the circuits could be preserved for scanning electron microscopic examination after the 4-hr study. MEASUREMENTS AND MAIN RESULTS: Platelet consumption was significantly reduced in both the heparinized and nonheparinized NO test groups when compared with the controls (p < .0001 and p < .0004, respectively). Platelet adhesion to the extracorporeal circuits was significantly reduced in the nonheparinized test circuits when compared with the controls (p < .05). Scanning electron microscopic examination of the circuits revealed that in the absence of heparin and in the presence of a NO-releasing surface, platelets retained their spherical nonactivated shape. CONCLUSIONS: The incorporation of NO into the surface of extracorporeal circuits reduces platelet consumption and eliminates the need for systemic heparinization in a rabbit model of extracorporeal circulation. PMID- 10809260 TI - Comparison of the renal effects of low to high doses of dopamine and dobutamine in critically ill patients: a single-blind randomized study. AB - OBJECTIVE: The renal effects of dopamine in critically ill patients remain controversial. Low-dose dobutamine has been reported to improve renal function. We compared the effects of various doses of dopamine and dobutamine on renal function in critically ill patients. DESIGN: Prospective, single-blind, randomized study. SETTING: University hospital, 19-bed multidisciplinary intensive care unit. PATIENTS: Twelve hemodynamically stable patients with mild nonoliguric renal impairment. INTERVENTIONS: Each patient randomly received four different doses of dopamine and dobutamine (placebo, 3, 7, and 12 microg/kg/min). Each infusion lasted for 4 hrs. Cardiac output and systemic hemodynamic variables were measured using a pulmonary arterial catheter at the beginning (HO) and the end (H4) of each infusion. The bladder was emptied at HO and H4 to determine urine volume and to collect samples. MEASUREMENTS AND MAIN RESULTS: The cardiac index increased significantly with both dopamine and dobutamine (p < .001). Mean arterial pressure (MAP) increased, with the maximum effect of 20% seen with 12 microg/kg/min dopamine infusion (p < .01). No change in MAP was seen with dobutamine. Dobutamine infusions did not change any renal variables. Conversely, all dopamine infusions significantly increased diuresis, creatinine clearance, and the fractional excretion of sodium (p < .01). Creatinine clearance increased from 61+/-16.9 (SD) mL/min to a maximum of 85.7+/-30 mL/min at the 7 microg/kg/min dose; fractional excretion of sodium increased from 0.26%+/-0.28% to a maximum of 0.62%+/-0.51% at the 12-microg/kg/min dose (p < .01). During dopamine infusions, there was a significant relationship between MAP and creatinine clearance (p = .018). CONCLUSIONS: At all doses studied, 4-hr infusions of dopamine significantly increased creatinine clearance, diuresis, and the fractional excretion of sodium in stable critically ill patients. Conversely, dobutamine did not modify these variables. Although the level of MAP might partially contribute to the improvement in renal variables, it is more likely that the activation of renal dopamine receptors played a prominent role. PMID- 10809261 TI - Interleukin-8 is increased in cerebrospinal fluid of children with severe head injury. AB - OBJECTIVE: To determine interleukin (IL)-8 concentrations in ventricular cerebrospinal fluid from children with severe traumatic brain injury (TBI). DESIGN: Prospective study. SETTING: University children's hospital. PATIENTS: Twenty-seven children hospitalized with severe TBI (Glasgow Coma Scale score < or =8), seven children with cerebrospinal fluid culture-positive bacterial meningitis, and twenty-four age-equivalent controls. INTERVENTIONS: Placement of an intraventricular catheter and continuous drainage of cerebrospinal fluid. MEASUREMENTS AND MAIN RESULTS: Median [range] cerebrospinal fluid IL-8 concentration in children with TBI (0-12 hrs) (4,452.5 [0-20,000] pg/mL) was markedly greater than that in controls (14.5 [0-250]) (p < .0001) and equivalent to concentrations in children with meningitis (5,300 [1,510-22,000] pg/mL) (p = .33). Cerebrospinal fluid IL-8 remained increased in children with severe TBI for up to 108 hrs after injury. Univariate logistic regression analysis demonstrated an association between cerebrospinal fluid IL-8 and child abuse (p = .07) and mortality (p = .01). Multivariate analysis demonstrated a strong, independent association between cerebrospinal fluid IL-8 and mortality (p = .01). CONCLUSIONS: The data are consistent with an acute inflammatory component of TBI in children and suggest an association between cerebrospinal fluid IL-8 and outcome after TBI. IL-8 may represent a potential target for anti-inflammatory therapy. PMID- 10809262 TI - Nosocomial pneumonia in patients undergoing heart surgery. AB - OBJECTIVE: To determine the risk factors related to the presence of postsurgical nosocomial pneumonia (NP) in patients who had undergone cardiac surgery. DESIGN: A case-control study. SETTING: Postcardiac surgical intensive care unit at a university center. PATIENTS: A total of 45 patients with NP and 90 control patients collected during a 4-yr period. INTERVENTIONS: Pre-, intra-, and postoperative factors were collected and compared between two groups of patients (cases vs. controls) to determine their influence on the development of NP. The diagnosis of NP was always microbiologically confirmed as pulmonary specimen brush culture of > or =10(3) colony-forming units/mL or positive blood culture/pleural fluid culture by the growth of identical microorganisms isolated at the lung. For each patient diagnosed with NP, we selected control cases at a ratio of 1:2. MEASUREMENTS AND MAIN RESULTS: The incidence of NP was 6.5%. Multivariate analysis found a probable association of the following variables with a greater risk for the development of NP: reintubation (adjusted odds ratio [AOR], 62.5; 95% confidence interval [CI], 8.1-480; p = .01); nasogastric tube (AOR, 19.7; 95% CI, 3.5-109; p = .01), transfusion of > or =4 units of blood derivatives (AOR, 12.8; 95% CI, 2-82; p = .01) and empirical treatment with broad spectrum antibiotics (AOR, 6.6; 95% CI, 1.2-36.8; p = .02). Culture results showed 13.3% of the NP to be of polymicrobial origin, whereas 77.3% of the microorganisms isolated were Gram-negative bacteria. The mortality (51 vs. 6.7%, p < .01) and the length of stay in the intensive care unit (25+/-14.8 days vs. 5+/-5 days, p < .01) were both greater in patients with NP. CONCLUSIONS: We conclude that the surgical risk factors, except the transfusion of blood derivatives, have little effect on the development of NP. Reintubation, nasogastric tubing, previous therapy with broad-spectrum antibiotics, and blood transfusion are factors most likely associated with NP acquisition. PMID- 10809263 TI - Spontaneous variability of cardiac output in ventilated critically ill patients. AB - OBJECTIVE: To define the magnitude of spontaneous cardiac output variability over time in sedated medical intensive care unit patients attached to a continuous cardiac output monitor, and to determine whether high level positive end expiratory pressure or inverse inspiratory-to-expiratory (I:E) ratio ventilation resulted in greater variability over time than low positive end-expiratory pressure with conventional I:E ratio ventilation. DESIGN: Prospective study. SETTING: Medical intensive care unit in a tertiary medical center. PATIENTS: A total of 22 hemodynamically stable acute respiratory failure patients with a pulmonary artery catheter inserted for hemodynamic monitoring INTERVENTIONS: After being sedated, patients were randomized ultimately to receive pressure control ventilation first at setting A (high positive end-expiratory pressure [15 cm H2O] with conventional I:E ratio [1:2]) and then at setting B (low positive end-expiratory pressure [5 cm H2O] with inverse I:E ratio [2:1]), or vice versa, and then at setting C (low positive end-expiratory pressure [5 cm H2O] with conventional I:E ratio [1:2]). Each ventilation setting period lasted 1 hr. MEASUREMENTS AND MAIN RESULTS: Cardiac output (CO) was measured continuously. The continuous CO value displayed was updated every 30-60 secs. The updated value reflected an average of the previous 3-6 mins. The coefficient of variation (CV) of CO for each setting in each patient was calculated to represent the spontaneous variability. The mean CO+/-SD and CV of each setting was 5.7+/-1.8 L/min and 4.4% for setting A, 5.6+/-1.5 L/min and 4.6% for setting B, and 5.9+/ 1.7 L/min and 4.8% for setting C. Analysis of variance revealed no significant differences between the CVs of the three settings. The 95% confidence interval for the COs for each setting was approximately the mean CO+/-0.1 x mean CO measured. CONCLUSIONS: In critically ill sedated medical intensive care unit patients with stable hemodynamics, the spontaneous variability of cardiac output over time was not significant. High positive end-expiratory pressure (15 cm H2O) and inverse ratio ventilation (2:1) did not contribute to increased spontaneous variability of cardiac output. PMID- 10809264 TI - Strong vasopressor support may be futile in the intensive care unit patient with multiple organ failure. AB - OBJECTIVE: The aim of the study was to determine the prognosis in patients who needed norepinephrine treatment in our institution in relation to the degree of organ failure and the evolution of the disease process. DESIGN: Retrospective case note analysis of outcome of those patients who needed norepinephrine according to our institutional regimen. PATIENTS: A total of 100 consecutive patients admitted to our 31-bed medical-surgical intensive care unit (ICU) who were treated with norepinephrine for severe hypotension and evidence of end-organ hypoperfusion unresponsive to both fluid resuscitation and dopamine treatment at 20 microg/kg/min. MEASUREMENTS: The degree of organ dysfunction at the time of starting norepinephrine treatment was assessed by the sequential organ failure assessment (SOFA) score. The time before starting norepinephrine treatment was defined as the time elapsed between ICU admission and that of starting norepinephrine administration. The patients were defined as survivors or nonsurvivors according to their ICU outcome. RESULTS: There were relationships between mortality and the degree of organ dysfunction and mortality and the duration of ICU stay before starting norepinephrine treatment. The mortality rate was 100% in the 30 patients with a total SOFA score of >12 and a delay before starting norepinephrine treatment of >1 day. The mortality rate of the other patients was 63%. The lowest mortality was seen in patients with lower SOFA scores and early norepinephrine administration after admission. CONCLUSIONS: Both the time of starting norepinephrine treatment after admission to the ICU and the degree of organ dysfunction have an important bearing on subsequent outcome. Although norepinephrine may be a lifesaving catecholamine in some cases, its administration to patients who have already developed multiple organ failure during their stay in the ICU is associated with a poor outcome. PMID- 10809265 TI - Relationship between procalcitonin plasma levels and severity of injury, sepsis, organ failure, and mortality in injured patients. AB - OBJECTIVE: To compare procalcitonin (PCT) plasma levels of injured patients with the incidence and severity of systemic inflammatory response syndrome (SIRS), infection, and multiple organ dysfunction syndrome (MODS) and to assess the predictive value of PCT for these posttraumatic complications. DESIGN: Retrospective study comparing patients with mechanical trauma in terms of severity of injury, development of infectious complications, and organ dysfunctions. SETTING: Level I trauma center with emergency room, intensive care unit, and research laboratory. PATIENTS: Four hundred five injured patients with an Injury Severity Score of > or =9 points were enrolled in this study from January 1994 to February 1996. INTERVENTIONS: Blood samples were collected on the day of admission and on days 1, 3, 5, 7, 10, 14, and 21 thereafter. MEASUREMENTS AND MAIN RESULTS: We determined PCT serum levels using a specific immunoluminometric assay. We retrospectively evaluated the occurrence of SIRS, sepsis, and MODS using patients' charts. Mechanical trauma led to increased PCT plasma levels dependent on the severity of injury, with peak values on days 1 and 3 (p < .05) and a continuous decrease within 21 days after trauma. Patients who developed SIRS demonstrated a significant (p < .05) increase of peak PCT plasma levels compared with patients without SIRS. The highest PCT plasma concentrations early after injury were observed in patients with sepsis (6.9+/-2.5 ng/mL; day 1) or severe MODS (5.7+/-2.2 ng/mL; day 1) with a sustained increase (p < .05) for 14 days compared with patients with an uneventful posttraumatic course (1.1+/-0.2 ng/mL). Moreover, these increased PCT plasma levels during the first 3 days after trauma predicted (p < .0001; logistic regression analysis) severe SIRS, sepsis, and MODS. CONCLUSIONS: These data indicate that PCT represents a sensitive and predictive indicator of sepsis and severe MODS in injured patients. Routine analysis of PCT levels seems to aid early recognition of these posttraumatic complications. Thus, PCT may represent a useful marker to monitor the inflammatory status of injured patients at risk. PMID- 10809266 TI - Comparison of methylene blue, riboflavin, and N-acetylcysteine for the reduction of nitric oxide-induced methemoglobinemia. AB - OBJECTIVE: To investigate the treatment of nitric oxide (NO)-induced methemoglobinemia by methylene blue (MB), riboflavin, and N-acetylcysteine (NAC) in vitro. DESIGN: Prospective, controlled in vitro study. SETTING: Research laboratory in a university hospital. PARTICIPANTS: Five healthy volunteers. INTERVENTIONS: Generation of 16% to 18% of methemoglobin in red blood cells by NO and subsequent addition of MB, riboflavin, or NAC. Simultaneous NO (32 ppm) and MB or riboflavin exposure of red blood cells. Induction of 14% to 18% of methemoglobin in red blood cells by NO, subsequent addition of MB or riboflavin, and further incubation with NO (80 ppm). MEASUREMENTS AND MAIN RESULTS: After discontinuation of NO, mean half-life for methemoglobin was significantly reduced by MB from 356 mins (controls) to 5 mins (10 microM) in a dose-dependent manner (p < .001). NAC did not alter the half-life for methemoglobin, and a reduction from 356 to 168 mins was seen for 120 microM riboflavin (p < .001). Methemoglobin formation after 3 hrs of NO exposure was 4.3%+/-0.7% in controls and 0.3%+/-0.1% with 10 microM MB (p < .001); 1 microM MB attenuated methemoglobin formation to 1.9%+/-0.1% (p < .01). With riboflavin (120 microM), methemoglobin was 2.2%+/ 0.5% vs. 3.2%+/-0.6% in controls (p < .001). In the presence of high methemoglobin concentrations, further methemoglobin formation was inhibited by 1 and 10 microM MB (p < .001) and attenuated by 0.1 microM MB (p < .001) but not by riboflavin. CONCLUSIONS: In vitro, NO-induced methemoglobin formation is significantly decreased by medium (1 microM) and high (10 microM) concentrations of MB and partially by high riboflavin concentrations (120 microM). NAC and low concentrations of riboflavin do not alter methemoglobin formation. PMID- 10809267 TI - Comparison of direct examination of three types of bronchoscopy specimens used to diagnose nosocomial pneumonia. AB - OBJECTIVE: To compare direct examination of bronchial aspirate and plugged telescopic catheter specimens (PTC) with infected cell counts in bronchoalveolar lavage (BAL) specimens for the diagnosis of nosocomial pneumonia. DESIGN: Prospective study of critically ill patients. SETTING: Intensive care unit in a university hospital. PATIENTS: A total of 64 patients hospitalized for >48 hrs with suspected nosocomial pneumonia. INTERVENTIONS: Fiberoptic bronchoscopy with bronchial aspirate and quantitative protected specimen brush, PTC, and BAL cultures. PTC and bronchial aspirate specimens were Gram-stained. BAL specimens for infected cell counts were examined as described previously in the literature. MEASUREMENTS AND MAIN RESULTS: Nosocomial pneumonia was diagnosed by the medical staff based on all available clinical, radiologic, laboratory test, and microbiological data and on the course before and after appropriate therapy. A total of 71% of patients were ventilated, and 70.1% were receiving antibiotics. Nosocomial pneumonia was diagnosed in 54% of the cases. On direct examination, sensitivity (Se) and specificity (Sp) of bronchial aspirate specimens were Se, 82% and Sp, 60%; of BAL with 5% infected cells, Se, 56% and Sp, 100%; of BAL with 3% infected cells, Se, 74% and Sp, 96%; of PTC specimens, Se, 65% and Sp, 76%; and of PTC specimens plus BAL with 3% infected cells, Se, 83% and Sp, 78%. BAL with 3% infected cells was significantly better for predicting nosocomial pneumonia than direct examination of bronchial aspirate or PTC specimens (p = .0012). When the BAL showed 3% infected cells, neither direct examination of bronchial aspirate nor direct examination of PTC specimens was useful (p = .24 and p = .38, respectively). Combined use of direct examination of PTC specimens plus BAL with 3% infected cells markedly improved sensitivity. The total cost of each procedure was taken into account for the final evaluation. CONCLUSIONS: Our data suggest that BAL with 3% infected cells is currently the only test whose predictive value for nosocomial pneumonia is sufficiently high to be of use for guiding the initial choice of antimicrobial class while waiting for quantitative culture results. PMID- 10809268 TI - Acute detoxification of opioid-addicted patients with naloxone during propofol or methohexital anesthesia: a comparison of withdrawal symptoms, neuroendocrine, metabolic, and cardiovascular patterns. AB - OBJECTIVE: Mu-Opioid receptor blockade during general anesthesia is a new treatment for detoxification of opioid addicted patients. We assessed catecholamine plasma concentrations, oxygen consumption, cardiovascular variables, and withdrawal symptoms after naloxone and tested the hypothesis that variables are influenced by the anesthetic administered during detoxification. DESIGN: Prospective randomized clinical study. SETTING: Intensive care unit of a university hospital and psychiatric ward. PATIENTS: Twenty-five mono-opioid addicted patients with mild to moderate systemic disease (ASA II classification) in a methadone substitution program. INTERVENTION: General anesthesia with either propofol (129+/-7 microg x kg(-1) x min(-1), mean +/- SEM) or methohexital (74+/ 14 microg x kg(-1). min(-1)), mu-opioid receptor blockade by naloxone in a stepwise fashion (increasing doses of 0.4 mg, 0.8 mg, 1.6 mg, 3.2 mg, and 6.4 mg at 15 min intervals followed by 0.8 mg x hr(-1) for 24 hrs) and naltrexone 50 mg x day(-1) orally for > or =4 wks. Clonidine was started 180 mins after the first naloxone dose and its infusion rate was individually adjusted to mitigate withdrawal symptoms during weaning and after extubation. MEASUREMENTS AND MAIN RESULTS: During propofol and methohexital anesthesia, naloxone induced a 30-fold increase in epinephrine and a significant three-fold increase in norepinephrine plasma concentrations without a significant difference between groups. This increase in catecholamine plasma concentrations was associated with increased oxygen consumption and marked cardiovascular stimulation with both anesthetics, as shown by increased cardiac index, heart rate, and systolic atrial pressure whereas diastolic pressure remained unchanged. Patients receiving propofol could be extubated significantly earlier after discontinuation of the anesthetics. Although the maximum degree of withdrawal symptoms (Short Opioid Withdrawal Scale) on the day after detoxification was similar with both anesthetics, subsequent withdrawal symptoms decreased significantly more rapidly after propofol anesthesia. CONCLUSIONS: Naloxone treatment, in opioid-addicted patients, induced a marked increase in catecholamine plasma concentrations, metabolism, and cardiovascular stimulation during anesthesia with both propofol and methohexital. Although both anesthetics appear suitable for detoxification treatment, the use of propofol is associated with earlier extubation and, surprisingly, a shortened period of long-term withdrawal symptoms during detoxification. PMID- 10809269 TI - Calcitonin precursors are reliable markers of sepsis in a medical intensive care unit. AB - OBJECTIVE: The diagnosis of infection in critically ill patients is challenging because traditional markers of infection are often misleading. For example, serum concentrations of calcitonin precursors are increased in patients with infections. However, their predictive accuracy for the diagnosis of sepsis in unselected patients in a medical intensive care unit (ICU) is unknown. Therefore, we compared the usefulness of serum concentrations of calcitonin precursors, C reactive protein, interleukin-6, and lactate for the diagnosis of sepsis in consecutive patients suffering from a broad range of diseases with an anticipated stay of > or =24 hrs in a medical ICU. DESIGN: Prospective cohort study. SETTING: Medical intensive care unit in a university medical center. PATIENTS: 101 consecutive critically ill patients. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: Blood samples were collected at various time points during the course of the disease. Systemic inflammatory response syndrome, sepsis, severe sepsis, and septic shock were diagnosed according to standardized criteria, and patients were reclassified daily without prior knowledge of the serum concentrations of calcitonin precursors or interleukin-6. At admission, 99% of the patients had systemic inflammatory response syndrome, 53% had sepsis, and 5% developed sepsis during their stay in the ICU. Calcitonin precursors, C-reactive protein, interleukin-6, and lactate levels increased with the severity of infection (p < .01, one-way analysis of variance). In a receiver operating characteristic curve analysis, calcitonin precursors were found to be the most reliable laboratory variable for the diagnosis of sepsis as compared with C-reactive protein, interleukin-6, and lactate (p < .01, for each comparison). Calcitonin precursor concentrations of >1 ng/mL had sensitivity of 89% and specificity of 94% for the diagnosis of sepsis. High serum concentrations of calcitonin precursors were associated with poor prognosis (p = .01). CONCLUSIONS: In a medical ICU, serum calcitonin precursor concentrations are more sensitive and are specific markers of sepsis as compared with serum C-reactive protein, interleukin-6, and lactate levels. PMID- 10809270 TI - Early identification of patients at risk for symptomatic vasospasm after aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: To develop a scheme for early identification of individuals at risk for symptomatic vasospasm after subarachnoid hemorrhage (SAH). DESIGN: Analysis of prospectively collected data from the placebo-treated group in a multicenter clinical trial. SETTINGS: Fifty-four neurosurgical centers in North America. MEASUREMENTS AND MAIN RESULTS: We identified independent predictors of symptomatic vasospasm using stepwise logistic regression analysis from demographic, clinical, laboratory, and neuroimaging characteristics of the participants. We developed a scoring system (symptomatic vasospasm risk index) based on a combination of these predictors. Out of 283 patients in the analysis (all treated with oral nimodipine), 93 (33%) developed symptomatic vasospasm within 14 days after SAH. There were four independent predictors of symptomatic vasospasm: thickness of subarachnoid clot on computed tomographic scan (odds ratio [OR], 4.1; 95% confidence interval [CI], 1.8-10.0); early rise in middle cerebral artery mean flow velocity (MCA-MFV), defined as a value > or =110 cm/sec recorded on or before post-SAH day 5 (OR, 1.9; 95% CI, 1.1-3.3), Glasgow Coma Scale score <14 (OR, 1.8; 95% CI, 1.1-3.1); and rupture of anterior cerebral or internal carotid artery aneurysm (OR, 1.9; 95% CI, 1.0-3.4). The probability of identifying patients who would develop symptomatic vasospasm (percentage of area under receiver operating characteristics curve +/- SEM) was higher with symptomatic vasospasm risk index (68%+/-8%) compared with thickness of clot (62%+/-8%; p = .08) or MCA-MFV (45%+/-7%, p < .05) criteria alone. CONCLUSIONS: Patients at high risk for symptomatic vasospasm can be identified early in the course of SAH using a risk index. A risk index based on a combination of variables may represent a predictive paradigm superior to conventionally used criteria based on clot thickness or MCA-MFV criteria. PMID- 10809271 TI - High prevalence of hyperhomocysteinemia in critically ill patients. AB - OBJECTIVE: To test the hypothesis that the prevalence of hyperhomocysteinemia is increased in critically ill patients and correlates with disease severity and mortality in these patients. DESIGN: A prospective study. SETTING: Three medical intensive care units at the University of vienna Medical School serving both medical and surgical patients. PATIENTS: All consecutive admissions (n = 56) during a period of 4 wks. A total of 112 age- and gender-matched healthy individuals constituted the control group. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood samples were drawn within 24 hrs after admission for analysis of total homocysteine (tHcy), folate, vitamin B6 levels, and vitamin B12 levels as well as to identify the 677C-->T polymorphism in the gene coding for the enzyme 5,10-methylenetetrahydrofolate reductase. Acute Physiology and Chronic Health Evaluation III scores at admission and 24 hrs after admission as well as 30-day survival were documented in all patients. Hyperhomocysteinemia was more prevalent in critically ill patients (16.1%; 95% confidence interval, 7.6% to 28.3%) compared with age- and gender-matched healthy individuals (5.4%; 95% confidence interval, 2.0% to 11.3%; chi-square test; p = .022). There was no difference in tHcy plasma concentrations in the first 24 hrs after admission to an intensive care unit between survivors and nonsurvivors. The 5,10 methylenetetrahydrofolate reductase 677C-->T polymorphism had no influence on tHcy levels and survival of intensive care unit patients. CONCLUSIONS: The prevalence of hyperhomocysteinemia is increased in critically ill patients compared to age- and gender-matched healthy individuals. The clinical significance of this finding remains to be determined. PMID- 10809272 TI - Cerebral perfusion, cardiac output, and arterial pressure in patients with fulminant hepatic failure. AB - OBJECTIVE: To evaluate whether changes in cardiac output influence cerebral perfusion directly. In fulminant hepatic failure, the circulation is characterized by wide variations in cardiac output and cerebral blood flow (CBF). DESIGN: A retrospective, interindividual analysis of CBF and cardiac output (part 1) and a prospective evaluation of cerebral perfusion, cardiac output, and arterial pressure during norepinephrine infusion (part 2). SETTING: A four-bed specialist liver failure unit. PATIENTS AND INTERVENTIONS: Twenty patients with fulminant hepatic failure (median age, 43 yrs; range, 17-54; 13 women) maintained on mechanical ventilation (Paco2, 33 torr [4.40 kPa]; range, 26-36 torr [3.47 4.80 kPa) after development of hepatic encephalopathy, stages 3 to 4, had mean arterial pressure (MAP) and cardiac output determined by radial and pulmonary artery catheters. Cerebral perfusion was measured by the 133Xenon clearance technique (n = 8) and by transcranial Doppler sonography, which was used to measure mean flow velocity (Vmean). CBF and Vmean in patients with high cardiac output (>9 L/min) were compared with those with normal or low cardiac output. In the second part of the study, cerebral autoregulation was evaluated by concomitant measurement of Vmean, cardiac output, and MAP during norepinephrine infusion in nine patients. MEASUREMENTS AND MAIN RESULTS: Median cardiac output was 8.5 L/min (range, 3.2-17.3), CBF was 33 mL/100 g/min (12-77 g/min), and Vmean was 45 cm/sec (22-65 cm/sec). In patients with elevated cardiac output, MAP, Vmean, and CBF were similar compared with patients with normal cardiac output. Neither CBF nor Vmean correlated to cardiac output. During norepinephrine infusion, Vmean increased from 49 cm/sec (34-69 cm/sec) to 63 cm/sec (58-90 cm/sec; p < .05), as MAP increased from 75 mm Hg (54-105 mm Hg) to 97 mm Hg (90 128 mm Hg). On average, cardiac output remained unchanged at 5.7 L/min (range, 3.2-17.3), as it increased in five patients and decreased in four patients. The change in Vmean was related to MAP (r2 = .76; p < .01) but not to cardiac output (r2 = .01). CONCLUSION: This study shows that CBF correlates to arterial pressure rather than to cardiac output in patients with fulminant hepatic failure. The presence of pressure-passive cerebral circulation stresses the importance of strict cardiovascular control in securing continuous and sufficient cerebral oxygenation and in avoiding the development of cerebral hyperemia and cerebral edema. PMID- 10809273 TI - The effect of phosphodiesterase III inhibitors on human neutrophil function. AB - OBJECTIVES: Neutrophils play an important role in ridding the body of bacteria and cellular debris. Several neutrophil functions are thought to be regulated by inotropes that increase cellular levels of cyclic adenosine monophosphate, including phosphodiesterase (PDE) inhibitors. We have investigated the effect of amrinone, milrinone, and olprinone, type III PDE (PDE-III) inhibitors, on several human neutrophil functions. DESIGN: Prospective in vitro study. SETTING: Academic research laboratory. SUBJECTS: Neutrophils isolated from 12 healthy adult volunteers. INTERVENTIONS: We measured chemotaxis, phagocytosis, reactive oxygen species production, intracellular calcium ion concentration, and cyclic adenosine monophosphate levels in neutrophils in the absence and the presence (at clinically relevant concentrations, 10 times, and 100 times those concentrations) of amrinone, milrinone, or olprinone. We also measured reactive oxygen species production under the same condition in a xanthine-xanthine oxidase system MEASUREMENTS AND MAIN RESULTS: None of the PDE-III inhibitors impaired neutrophil chemotaxis or phagocytosis. Amrinone at clinically relevant or higher concentrations and milrinone at high concentrations reduced superoxide, hydrogen peroxide, and hydroxyl radical levels in neutrophils and in the xanthine-xanthine oxidase system. Olprinone did not have those effects, and none of the PDE-III inhibitors had an effect on intracellular calcium ion concentration or cyclic adenosine monophosphate production in neutrophils stimulated by a chemotactic factor. CONCLUSIONS: The ability of amrinone to scavenge reactive oxygen species at clinically relevant concentrations while not affecting neutrophil function suggests that the PDE inhibitor can be used without detriment in severely ill patients. PMID- 10809274 TI - A picture archival and communication system shortens delays in obtaining radiographic information in a medical intensive care unit. AB - OBJECTIVE: To assess whether variables such as unit occupancy and aggregate severity of illness that reflect increased work demands on physicians in medical intensive care units (MICU) are associated with increased delays in their obtaining information about nonroutine chest radiographic examinations. To determine whether the presence of a picture archiving and communication system (PACS) workstation in the MICU shortens those delays. DESIGN: A prospective cohort study stratified for presence or absence of PACS. SETTING: MICU of a university hospital. PATIENTS: A total of 118 patients admitted to the MICU who had nonroutine bedside chest radiographs. MEASUREMENTS AND MAIN RESULTS: Multivariate analyses were conducted to determine how unit occupancy, patient acuity, the time of day the examination was taken, and the presence of a PACS workstation influenced the time from radiographic examination completion to the time when MICU physicians first obtained image information. In a multivariate analysis, patient acuity, unit occupancy, the aggregate level of severity of illness in the study cohort, whether the examination was taken at night or day, and the presence of a PACS workstation were significant predictors of the elapsed time from examination completion until review by MICU physicians. Without the PACS workstation, higher occupancy, higher aggregate severity of illness, and examinations taken during the day were associated with longer delays. Overall, the multivariate analysis showed a 24-min decrease in the elapsed time to obtain information during periods with the PACS workstation compared with periods without the workstation (p = .03). CONCLUSIONS: A PACS workstation significantly decreased the delays in obtaining image information that occurred with high unit occupancy and high aggregate severity of illness and may improve unit efficiency under conditions of high physician workload. PMID- 10809275 TI - Outcome of bone marrow transplantation patients requiring mechanical ventilation. AB - OBJECTIVE: To identify outcome predictors in bone marrow transplantation (BMT) patients admitted to the intensive care unit (ICU) of The University of Texas M. D. Anderson Cancer Center who required endotracheal intubation and mechanical ventilation. DESIGN: Retrospective, comparative study. SETTING: A 16-bed medical intensive care unit in a university teaching cancer center. PATIENTS: The records of 60 consecutive BMT patients who developed respiratory failure requiring mechanical ventilation were reviewed. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The most frequent complication leading to respiratory failure was pneumonia (41%) followed by diffuse alveolar hemorrhage (37%). Eighteen percent of the patients were extubated and discharged from the ICU, but only 5% were alive at 6 months. Graft vs. host disease was a predictor of a poor outcome (p < .05). Breast cancer as an underlying disease and pulmonary edema as a complication were favorable predictive factors (p < .05). Five of 26 patients with diffuse alveolar hemorrhage and four of 33 patients with pneumonia survived. We found no relationship between survival and age, gender, BMT type, or Acute Physiology and Chronic Health Evaluation II score. Prolonged mechanical ventilation (> or =15 days) and late development of respiratory failure (>30 days after BMT) were associated with poor prognosis. CONCLUSIONS: The ICU survival rate of BMT patients who developed pulmonary complications and required mechanical ventilation was 18%. Prognostic factors were described identifying patients with a substantial survival rate as well as those in whom mechanical ventilation was futile. PMID- 10809276 TI - Continuous calculation of intratracheal pressure in the presence of pediatric endotracheal tubes. AB - OBJECTIVE: To measure the pressure-flow relationship of pediatric endotracheal tubes (ETTs) in trachea models, to mathematically describe this relationship, and to evaluate in trachea/lung models a method for calculation of pressure at the distal end of the ETT (Ptrach) by subtracting the flow-dependent pressure drop across the ETT from the airway pressure measured at the proximal end of the ETT. DESIGN: Trachea models and trachea/lung models. SETTING: Research laboratory in a university medical center. INTERVENTIONS: The pressure-flow relationship of pediatric ETTs (inner diameter, 2.5-6.5 mm) was determined using a physical model consisting of a tube connector, an anatomically curved ETT, and an artificial trachea. The model was ventilated with sinusoidal gas flow (12-60 cycles/min). The coefficients of an approximation equation considering ETT resistance and inertance were fitted separately to the measured pressure-flow curves for inspiration and expiration. Calculated Ptrach was compared with directly measured Ptrach in mechanically ventilated physical trachea/lung models. MEASUREMENTS AND MAIN RESULTS: The pressure-flow relationship was considerably nonlinear and showed hysteresis around the origin caused by the inertia of accelerated gas. ETT inertance ranged from 0.1 to 0.4 cm H2O/L x sec2 (inner diameter, 6-2.5 mm). The abrupt change in cross-sectional area at the tube connector caused an inspiration to-expiration asymmetry. Calculated and measured Ptrach were within +/- 1 cm H2O. Correspondence between measured and calculated Ptrach is improved even further when the ETT inertance is taken into account. CONCLUSIONS: Ptrach can continuously be monitored in the presence of pediatric ETT by combining ETT coefficients and the flow and airway pressure continuously measured at the proximal end of the ETT. PMID- 10809277 TI - Dependency of cerebral blood flow on mean arterial pressure in patients with acute bacterial meningitis. AB - OBJECTIVE: Patients with acute bacterial meningitis are often treated with sympathomimetics to maintain an adequate mean arterial pressure (MAP). We studied the influence of such therapy on cerebral blood flow (CBF). DESIGN: Prospective physiologic trial. SETTING: The Department of Infectious Diseases, Copenhagen University Hospital, Denmark. PATIENTS: Sixteen adult patients with acute bacterial meningitis. INTERVENTION: Infusion of norepinephrine to increase MAP. MEASUREMENTS: During a rise in MAP induced by norepinephrine infusion, we measured relative changes in CBF by transcranial Doppler ultrasonography of the middle cerebral artery, recording mean flow velocity (Vmean), and by the arterial to jugular oxygen saturation difference. In 10 out of 16 patients, serial measurements were performed until recovery or death. Individual autoregulation curves were analyzed by a computer program. Autoregulation was classified as impaired if Vmean increased by >10% per 30 mm Hg increase in MAP and if no lower limit of autoregulation was identified by the computer program; otherwise, autoregulation was classified as preserved. MAIN RESULTS: Initially, Vmean increased from a median value of 46 cm/sec (range, 30-87 cm/sec) to 63 cm/sec (33 105 cm/sec) (p < .0001), and arterial to jugular oxygen saturation difference decreased from 0.28 (0.16-0.51) to 0.21 (0.08-0.39) (p < .001) when MAP was raised from 69 mm Hg (55-102 mm Hg) to 110 mm Hg (93-129 mm Hg). CBF autoregulation was restored in eight of ten patients undergoing serial examination after 7 (range, 2-10) days. Six of these patients had an uncomplicated course, one had a protracted recovery, and one died. Autoregulation was not restored in two patients; one died and one had a protracted recovery. CONCLUSION: In patients in the early phase of acute bacterial meningitis, CBF autoregulation is impaired. With recovery from meningitis, the cerebral vasculature regains the ability to maintain cerebral perfusion at a constant level despite variations in MAP. PMID- 10809278 TI - Efficiency of chest computed tomography in critically ill patients with multiple traumas. AB - OBJECTIVE: The efficiency of secondary thoracic computed tomography (TCT) in critically ill patients with multiple traumas was assessed by comparison of TCT with chest radiograph findings. The subsequent therapeutic consequences based on the additional information of TCT were evaluated. SETTING: A six-bed trauma intensive care unit in a university hospital. DESIGN: Prospective, descriptive study. PATIENTS AND INTERVENTIONS: One hundred one computed tomographic (CT) examinations (mean, 2.6 per patient; range, 1-10) were performed in 39 patients, fulfilling the following indications for TCT: a) sepsis with suspected pulmonary focus (n = 41); b) deterioration of pulmonary gas exchange (n = 35); c) guiding the duration of intermittent prone positioning (n = 25). The information provided by TCT was compared with corresponding chest radiographs (CXR). Therapeutic consequences drawn after TCT were compared with the additional diagnostic information of TCT. The change of therapy was documented that would not have been undertaken or may have been delayed had TCT evaluation not been used. RESULTS: TCT was significantly superior to CXR in detecting pneumothoraces, pleural effusions, and pulmonary abscesses. Furthermore, a significantly higher accuracy regarding pulmonary densities was found. Subsequent therapeutic interventions ensued from 85 (84.2%) CT scans. After TCT, intermittent prone positioning was initiated in 31 patients, chest tubes were inserted in 16 patients, and intermittent prone positioning was terminated in 13 patients and was continued in 12 patients. Eleven thoracotomies were performed because of the TCT findings. The described therapeutic interventions were based on abnormalities seen on CT scans but were not evident in CXR in 58 patients (57.4%). Significant information that influenced therapeutic concepts was obtained in 66% (n = 23) of patients with pulmonary deterioration of gas exchange, in 61% (n = 25) of patients with sepsis, and in 40% (n = 10) of patients to guide the duration of intermittent prone positioning. Thoracotomy and specific drainage by tube thoracostomy was always dependent on the findings of TCT. CONCLUSION: Performed under the above displayed defined indications, TCT had an overall efficiency of 57%. It provided an increased sensitivity for intrathoracic lesions and a more comprehensive diagnosis of chest abnormalities. PMID- 10809279 TI - Induction of systemic serum procalcitonin and cardiocirculatory reactions after isolated limb perfusion with recombinant human tumor necrosis factor-alpha and melphalan. AB - OBJECTIVES: Isolated, hyperthermic limb perfusion (ILP) with recombinant human tumor necrosis factor-alpha (rhTNF-alpha) and melphalan is a highly effective treatment for locoregional metastases of malignant melanoma and for advanced soft tissue sarcoma of the limb. The major systemic side effects are characterized by the induction of a systemic inflammatory response syndrome (SIRS). Procalcitonin (PCT), a serum marker of bacterial sepsis, was investigated with respect to its role in SIRS after ILP. SETTING: University surgical oncology division with an integrated eight-bed intensive care unit. PATIENTS: Thirty-seven patients were treated by ILP with rhTNF-alpha and melphalan (n = 26) or with cytostatics alone (n = 11) for soft tissue sarcoma or malignant melanoma. INTERVENTIONS: The course of serum PCT, interleukin (IL)-6, and IL-8 was analyzed intra- and postoperatively. Hemodynamic variables including heart rate, mean arterial pressure, cardiac index, pulmonary arterial pressure, pulmonary capillary occlusion pressure, and pulmonary and systemic vascular resistance were recorded in parallel. MEASUREMENTS AND MAIN RESULTS: PCT was significantly elevated over baseline after ILP with a maximum between 8 hrs (peak level 16.0+/-18.8 (SD) ng/mL) and 36 hrs (13.8+/-15.7 ng/mL) (p < .001). The increase in serum PCT was significantly more pronounced after ILP with rhTNF-alpha/melphalan than after ILP with cytostatics alone (p < .001). IL-6 and IL-8 were also significantly increased after ILP (p = .001), reaching peak concentrations at 1 hr and 4 hrs postoperatively. Significant changes in heart rate, mean arterial pressure, cardiac index, and systemic vascular resistance were observed during and after ILP; however, PCT levels could not be correlated to these variables. Pulmonary arterial pressure, pulmonary capillary occlusion pressure, and pulmonary vascular resistance showed no significant changes. CONCLUSIONS: Serum procalcitonin is induced as part of the SIRS after ILP with rhTNF-alpha/melphalan. It may be induced directly by rhTNF-alpha or other cytokines, because serum peaks of IL-6 and IL-8 precede the peak of PCT. Because there is no correlation between serum levels of PCT and hemodynamic variables, this marker cannot be applied to assess the severity of SIRS reaction after ILP. PMID- 10809280 TI - Increased nuclear factor kappa B activation in critically ill patients who die. AB - OBJECTIVES: To determine nuclear factor kappa B (NF-kappa B) activation in mononuclear and neutrophils from critically ill patients and to compare NF-kappa B activation with circulating concentrations of interleukin (IL)-6, IL-8, and soluble intercellular adhesion molecule (sICAM)-1. DESIGN: Observational study. SETTING: University Teaching Hospital, eight-bed intensive care unit in northeast Scotland. PATIENTS: Ten patients admitted to the intensive care unit who fulfilled the criteria for systemic inflammatory response syndrome were studied at 0, 24, 48, and 72 hrs. Six healthy volunteers were also studied. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: NF-kappa B activation was significantly higher in patients compared to healthy volunteers in both neutrophils (p = .001) and mononuclear leukocytes (p = .013). In the six patients who survived to 96 hrs, the level of NF-kappa B activation in mononuclear cells remained constant (p = .9). However, in the four patients who died before 96 hrs, mononuclear cell NF-kappa B activation increased markedly and was significantly higher before death than in those who survived to 96 hrs (p = .0105). NF-kappa B activation in neutrophils similarly remained constant in patients who survived to 96 hrs (p = .4) but did not show the same increase before death. Circulating concentrations of IL-6, IL-8, and sICAM-1 were elevated but were unrelated to leukocyte NF-kappa B activation. CONCLUSIONS: We found NF-kappa B activation in mononuclear and neutrophils in patients with systemic inflammatory response syndrome, which increased markedly before death in mononuclear leukocytes and was not related to plasma IL-6, IL-8, and sICAM-1 concentrations. These data support the need for further study of the role of NF-kappa B activation in mortality from systemic inflammatory response syndrome and sepsis. PMID- 10809281 TI - Clinical experience with cerebral oximetry in stroke and cardiac arrest. AB - OBJECTIVE: To address the ability and reliability of the INVOS 3100A (Somanetics, Troy, MI) cerebral oximeter to detect cerebral desaturation in patients and the interpretation of cerebral oximetry measurements using the INVOS 3100A in stroke and cardiac arrest. DESIGN: Case reports of two patients. SETTING: Neurologic Intensive Care Unit of a University Hospital. PATIENTS: Two patients suffering occlusive strokes of the middle cerebral artery. One later suffered a cardiac arrest. RESULTS: The first case, a patient who suffered cardiac arrest while undergoing continuous cerebral oximetry, clearly demonstrated the ability of the INVOS 3100A to detect rapid tissue vascular oxyhemoglobin desaturation in the brain during circulatory arrest. In the second case, oximetry readings were obtained in a patient with a right internal carotid artery occlusion and an infarct in the middle cerebral artery territory. The circulation of the anterior cerebral artery (ACA) territory was intact. Stable xenon-computed tomography of local cerebral blood flow showed no perfusion in the infarct, and oximetry readings were between 60 and 65. In the border zone between the middle cerebral artery and the ACA, readings of 35 to 40 were obtained, and over the ACA territory, the readings were in the 60s. CONCLUSIONS: Oximetry by near infrared spectroscopy reflects the balance between regional oxygen supply and demand. In dead or infarcted nonmetabolizing brain, saturation may be near normal because of sequestered cerebral venous blood in capillaries and venous capacitance vessels and contribution from overlying tissue. In regionally or globally ischemic, but metabolizing brain, saturation decreases because oxygen supply is insufficient to meet metabolic demand. These observations are supported by previously reported "normal" readings in unperfused or dead brains. PMID- 10809282 TI - Safety and efficiency of metered dose inhaler delivery of salbutamol in the intubated rabbit. AB - OBJECTIVE: To determine the safety and efficiency of metered dose inhaler salbutamol delivered to the intubated rabbit. DESIGN: Prospective, comparative, five-group laboratory investigation. SETTING: Animal laboratory, Department of Nuclear Medicine. SUBJECTS: A total of 30 adult, anesthetized New Zealand White rabbits. INTERVENTIONS: Three groups of rabbits underwent tracheal intubation through a tracheostomy and received 5 puffs of 99mTcO4 salbutamol delivered at the elbow connector (group 1) or via a catheter with its distal tip positioned at the midpoint (group 2) or bevel of the endotracheal tube (group 3). No intervention was provided for the rabbits in the fourth group. A fifth group underwent tracheal intubation through the mouth and received salbutamol (5 puffs) delivered at the bevel of the endotracheal tube. MEASUREMENTS: Delivery efficiency was expressed as the ratio of radioactivity emitted from lungs and trachea to the total radioactivity of the administered dose. Histopathologic injury scores were assigned to each trachea or lung specimen. MAIN RESULTS: Delivery efficiency was 30 times greater in groups 3 and 5 (full catheter) than in group 1 (elbow). The injury scores were similar in all groups. CONCLUSION: We conclude that the increased efficiency obtained by administration of metered dose inhaler salbutamol at the distal tip of endotracheal tube is not necessarily associated with increased epithelial injury. PMID- 10809283 TI - Noninvasive monitoring of nonshunted pulmonary capillary blood flow in the acute respiratory distress syndrome. AB - OBJECTIVE: Noninvasive monitoring of nonshunted pulmonary capillary blood flow, using the alveolar amplitude response technique (AART) in a porcine model of the acute respiratory distress syndrome. DESIGN: Experimental animal study. SETTING: University center for animal experiments. INTERVENTIONS: In 12 mechanically ventilated pigs, the nonshunted pulmonary capillary blood flow was varied by means of lung lavages and the application of positive end-expiratory pressure. MEASUREMENTS AND MAIN RESULTS: Nonshunted pulmonary capillary blood flow was determined by AART. Cardiac output (determined by the thermodilution method) corrected for venous admixture was used for comparison (r2 varied between .58 and .94; p < .01). The trend in the development of nonshunted pulmonary capillary blood flow as measured with AART was in agreement with the trend detected by cardiac output corrected for venous admixture in 92% of all events. CONCLUSIONS: We conclude that AART can be used to monitor changes in nonshunted pulmonary capillary blood flow in cases of acute respiratory distress syndrome noninvasively and continuously. PMID- 10809284 TI - Combined effects of inhaled nitric oxide (iNO) and oxidant agents on the production of methemoglobinemia in newborn piglets. AB - OBJECTIVE: To investigate the effects of the association of inhaled nitric oxide (iNO) and oxidant drugs (acetaminophen, phytomenadione, and EMLA cream) on methemoglobinemia during the neonatal period. DESIGN: Prospective, randomized, experimental study. SETTING: University Experimental Pharmacology laboratory. SUBJECTS: Sixty newborn piglets weighing 1.5-2.0 Kg. INTERVENTIONS: Twelve groups of five piglets were anaesthetized, mechanically ventilated, and studied for 3 hrs. Eight groups received iNO (40 ppm or 80 ppm) alone or in association with a single intravenous dose of acetaminophen (120 mg/kg propacetamol), phytomenadione (5 mg vitamin K1) or EMLA cream (2.5 g) applied to the ventral lower abdomen for 3 hrs. Three other groups received, respectively, acetaminophen, phytomenadione, or EMLA cream without iNO. The last group (control group) received neither drugs nor iNO. MEASUREMENTS AND MAIN RESULTS: Methemoglobinemia was measured before the beginning of each experiment, 30 mins later, and every hour for 3 hrs. There was no significant difference in methemoglobinemia at any time between groups receiving acetaminophen (0.90%+/-0.12%), phytomenadione (0.88%+/-0.11%), or EMLA cream alone (0.97%+/-0.11%) and the control group (0.92%+/-0.12%). At 3 hrs, methemoglobinemia was slightly but significantly increased in group receiving iNO alone (1.04%+/-0.17% at 40 ppm iNO and 1.14%+/-0.16% at 80 ppm iNO; p < .05). Conversely, methemoglobinemia increased as a function of time in groups in which iNO was associated to drug administration and was significantly greater than the control group at 3 hrs (80 ppm iNO + acetaminophen, 2.80%+/-0.47%; 80 ppm iNO + phytomenadione, 2.38%+/-0.45%; 80 ppm iNO + EMLA cream, 2.33%+/-046%; p < .001). CONCLUSIONS: These results demonstrate that if oxidant drugs (acetaminophen, phytomenadione, or EMLA cream) did not increase blood methemoglobinemia in neonatal piglets, their association with iNO caused an increase in methemoglobin. Special care should be taken to monitor methemoglobinemia when iNO is combined to such drugs in newborn infants. PMID- 10809286 TI - Analysis of ketamine responses in the pulmonary vascular bed of the cat. AB - OBJECTIVE: To test the hypothesis that pulmonary vasodilator responses of ketamine are dependent on activation of L-type calcium channels, independent of synthesis of nitric oxide from L-arginine, activation of adenosine triphosphate sensitive potassium channels, and the release of cyclooxygenase products. DESIGN: Prospective study. SETTING: Research laboratory. SUBJECTS: Isolated lobar lung preparation, mongrel cats. INTERVENTIONS: In separate experiments, the effects of nicardipine; N omega-I-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase; glibenclamide, an adenosine triphosphate-sensitive potassium channel antagonist; and meclofenamate, a cyclooxygenase blocker, were investigated in the pulmonary vascular bed of the cat. The effects of these agents were evaluated on the pulmonary arterial responses of ketamine, acetylcholine, and isoproterenol during elevated tone conditions induced by the thromboxane A2 mimic, U46619 (Upjohn, Kalamazoo, MI). MEASUREMENTS: Lobar arterial perfusion pressure, systemic pressure, and left atrial pressure were continuously monitored, electronically averaged, and permanently recorded. MAIN RESULTS: Under elevated tone conditions in the isolated left lower lobe vascular bed of the cat, N omega-I-nitro-L-arginine methyl ester, glibenclamide, and meclofenamate had no significant effect on the vasodilator responses to ketamine. Nicardipine, in a dose that reduced significantly vasopressor effects to BAY K 8644, a calcium-channel opener, attenuated significantly vasodilator responses to ketamine, whereas the L-type calcium-channel blocker had no significant effects on responses to acetylcholine and to isoproterenol. CONCLUSIONS: These data show that ketamine has significant vasodilator activity in the pulmonary vascular bed of the cat. The present data also suggest that responses to ketamine during elevated tone conditions may in part be mediated by the activation of L-type calcium channels. PMID- 10809285 TI - Inhaled nitric oxide modulates leukocyte kinetics in the mesenteric venules of endotoxemic rats. AB - OBJECTIVE: to determine whether inhaled nitric oxide (NO) would alter leukocyte kinetics in the septic microvasculature. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Sprague Dawley rats. INTERVENTIONS: Rats were treated with either saline or endotoxin (10 mg/kg, iv) and then allowed to breathe either air or air plus NO (10 ppm). MEASUREMENTS AND MAIN RESULTS: After a 4-hr period, rolling, firm adhesion, and emigration of leukocytes and endothelial dysfunction were monitored in mesenteric venules by using intravital videomicroscopy. Compared with controls, endotoxemic rats exhibited a profound influx in mesenteric venule rolling leukocytes (55+/-17 vs. 70+/-19 leukocytes/min; p < .05), associated with a reduction of leukocyte rolling velocity (83+/-14 vs. 34+/-3 microm/sec; p < .05). In endotoxemic rats, venular endothelium leukocyte firm adhesion (1.15+/-0.32 vs. 4.08+/-0.96 leukocytes/ 100 microm; p < .05) and emigration (0.84+/-0.47 vs. 4.23+/-1.2 leukocytes/100 microm; p < .05) increased compared with controls. Inhaled NO had no effect on leukocyte kinetics in control rats. Inhaled NO significantly attenuated endotoxin-induced venular endothelium leukocyte adhesion (4.08+/-0.96 vs. 1.86+/-0.76 leukocytes/100 microm; p < .05) and emigration (4.23+/-1.2 vs. 1.68+/-0.72 leukocytes/100 microm; p < .05). Compared with control rats, macromolecular (FITC-dextran) vascular leakage, expressed as the perivenular/intravenular fluorescence intensity ratio, increased in endotoxemic rats (0.56+/-0.02 vs. 0.81+/-0.05; p < .01). Endotoxin-induced macromolecular vascular leakage increases were partially prevented by inhaled NO (0.66+/-0.01 vs. 0.56+/-0.02; p < .05). CONCLUSION: These observations suggest that inhaled NO reduces leukocyte adhesion and the degree of vascular permeability dysfunction in mesenteric venule of endotoxemic rats. PMID- 10809287 TI - Effects of vasopressin and epinephrine on splanchnic blood flow and renal function during and after cardiopulmonary resuscitation in pigs. AB - OBJECTIVE: To compare the effects of vasopressin versus epinephrine on splanchnic blood flow during and after cardiopulmonary resuscitation (CPR), and to evaluate the effects of these vasopressors on renal function in the postresuscitation phase. DESIGN: Prospective, randomized laboratory investigation using an established porcine CPR model with instrumentation for continuous measurement of splanchnic and renal blood flow. SETTING: University hospital experimental laboratory. SUBJECTS: A total of 12 anesthetized, 12- to 16-wk-old domestic pigs weighing 30-35 kg. INTERVENTIONS: After 4 mins of cardiac arrest, and 3 mins of CPR, 12 pigs were randomly assigned to receive either 0.4 units/kg vasopressin (n = 6) or 45 microg/kg epinephrine (n = 6). Defibrillation was performed 5 mins after drug administration; all animals were observed for 6 hrs after return of spontaneous circulation (ROSC). MEASUREMENTS AND MAIN RESULTS: Mean +/- SEM superior mesenteric artery blood flow was significantly (p < .05) lower after vasopressin compared with epinephrine at 90 secs after drug administration (13+/ 3 vs. 129+/-33 mL/min); at 5 mins after drug administration (31+/-18 vs. 155+/-39 mL/min); at 5 mins after ROSC (332+/-47 vs. 1087+/-166 mL/min); and at 15 mins after ROSC (450+/-106 vs. 1130+/-222 mL/min); respectively. Mean +/- SEM left renal and hepatic artery blood flow after ROSC was comparable in both groups ranging between 120-290 mL/min (renal blood flow), and 150-360 mL/min (hepatic blood flow), respectively. Median urine output after ROSC showed no difference between groups, and highest values (180-220 mL/hr) were observed in the first 60 mins after ROSC. Median calculated glomerular filtration rate showed no difference between groups with values ranging between 30 and 80 mL/min in the postresuscitation phase. Calculated fractional sodium excretion and osmolar relationship between urea and plasma indicated no evidence for renal tubular dysfunction. CONCLUSIONS: In the early postresuscitation phase, superior mesenteric blood flow was temporarily impaired by vasopressin in comparison with epinephrine. With respect to renal blood flow and renal function after ROSC, there was no difference between either vasopressor given during CPR. Vasopressin given during CPR did not result in an antidiuretic state in the postresuscitation phase. PMID- 10809288 TI - Effects on capillary fluid permeability and fluid exchange of albumin, dextran, gelatin, and hydroxyethyl starch in cat skeletal muscle. AB - OBJECTIVE: To evaluate the peripheral hemodynamic effects in a skeletal muscle in vivo model of the four commercially available colloid solutions, 20% human albumin, 6% dextran-70, 6% hydroxyethyl starch 200/0.5 (HES), and 3.5% urea linked gelatin. DESIGN: Controlled laboratory study. SETTING: University research laboratory. SUBJECTS: Fifteen adult cats. INTERVENTIONS: The isolated, autoperfused, and denervated calf muscles of the cat hindlimb were placed in a plethysmograph. The colloids tested were given intra-arterially to the muscles in increasing rates of clinically relevant doses. MEASUREMENTS AND MAIN RESULTS: Arterial blood flow, arterial and venous blood pressures, total vascular resistance, tissue volume changes, and capillary filtration coefficient were measured before, during, and after the colloid infusions. The altered capillary filtration coefficient reflects a change in capillary fluid permeability. The capillary filtration coefficient was decreased by albumin and dextran, not affected by HES, and increased by urea-linked gelatin. Albumin induced transcapillary fluid absorption, gelatin induced transcapillary filtration, and no transcapillary fluid exchange was observed with dextran and HES. After discontinuation of the infusions, HES and gelatin induced a rebound transcapillary filtration. No such effect was seen after dextran and albumin. All colloids increased muscle blood flow. CONCLUSION: We conclude that capillary fluid permeability is decreased by albumin and dextran, unchanged by HES, and increased by gelatin. This and the differences in the rebound effect may contribute to the differences in the plasma volume expanding properties of the respective colloid. The increased blood flow induced by the colloids was more an effect of reduced vascular tone than of lowered blood viscosity. PMID- 10809289 TI - Vasopressin decreases endogenous catecholamine plasma concentrations during cardiopulmonary resuscitation in pigs. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of vasopressin vs. saline placebo on catecholamine plasma concentrations during cardiopulmonary resuscitation (CPR). DESIGN: Prospective, randomized laboratory investigation by using an established porcine CPR model with instrumentation for measurement of hemodynamic variables, vital organ blood flow, and return of spontaneous circulation. SETTING: University hospital laboratory. SUBJECTS: Sixteen domestic pigs. INTERVENTIONS: After 15 mins of untreated cardiac arrest and 3 mins of CPR, 16 pigs were randomized to be treated with either 0.8 U/kg vasopressin (n = 8) or placebo (normal saline; n = 8). Arterial epinephrine and norepinephrine plasma concentrations were sampled at prearrest, after 1.5 mins of chest compressions, and at 1.5 mins and 5 mins after drug administration during CPR. MEASUREMENTS AND MAIN RESULTS: In comparison with placebo pigs at 1.5 and 5 mins after drug administration, animals resuscitated with vasopressin had significantly (p < .01) higher mean +/- SEM left ventricular myocardial (131+/-27 vs. 10+/-1 mL x mins( 1) x 100 g(-1) and 62+/-13 vs. 9+/-2 mL x mins(-1) x 100 g(-1)); total cerebral (90+/-8 vs. 14+/-3 mL x mins(-1) x 100 g(-1) and 51+/-4 vs. 12+/-2 mL x mins(-1) x 100 g(-1)); and adrenal gland perfusion (299+/-36 vs. 38+/-7 mL x mins(-1) x 100 g(-1) and 194+/-23 vs. 29+/-5 mL x mins(-1) x 100 g(-1)). Significantly lower mean +/- SEM epinephrine concentrations in the vasopressin pigs compared with the placebo group were measured 1.5 mins and 5 mins after drug administration, (24167+/-7919 vs. 80223+/-19391 pg/mL [p < .01] and 8346+/-1454 vs. 71345+/-10758 pg/mL [p < .01]). Mean +/- SEM norepinephrine plasma concentrations in the vasopressin animals in comparison with placebo were at 1.5 and 5 mins after drug administration significantly lower (41729+/-13918 vs. 82756+/-9904 pg/mL [p = .01] and 10642+/-3193 vs. 62170+/-8797 pg/mL [p < .01]). CONCLUSIONS: Administration of vasopressin during CPR resulted in significantly superior vital organ blood flow, but significantly decreased endogenous catecholamine plasma concentrations when compared with placebo. PMID- 10809290 TI - Effects of propofol on hemodynamic and inflammatory responses to endotoxemia in rats. AB - OBJECTIVE: To document the effects of propofol on the hemodynamic and inflammatory responses to endotoxemia in an animal model. DESIGN: Randomized, prospective laboratory study. SETTING: University experimental laboratory. SUBJECTS: Thirty-two male rats. INTERVENTIONS: The animals were randomly assigned to one of four groups: a) endotoxemia group (n = 8), which received intravenous Escherichia coli endotoxin (15 mg/kg over 2 mins); b) control group (n = 8), which was treated identically to the endotoxemia group except for the substitution of 0.9% saline for endotoxin; c) propofol group (n = 8), which was treated identically to the control group but also received propofol (10 mg/kg bolus, followed by infusion at 10 mg/kg/hr) immediately after the injection of 0.9% saline; and d) propofol-endotoxemia group (n = 8), which was treated identically to the endotoxemia group with the additional administration of propofol (10 mg/kg bolus, followed by infusion at 10 mg/kg/hr) immediately after endotoxin injection. MEASUREMENTS AND MAIN RESULTS: Hemodynamics, arterial blood gases, and acid-base status were recorded and the blood propofol concentrations and plasma cytokine concentrations were measured during the 5-hr observation. Microscopic findings of lung tissue for each group were obtained at necropsy. The systolic arterial pressure and heart rate of the propofol-endotoxemia group were similar to those of the endotoxemia group. The increases in the plasma cytokine (tumor necrosis factor, interleukin-6, and interleukin-10) concentrations, in the base deficit, and in the infiltration of neutrophils in the air space or vessel walls of the lungs were attenuated in the propofol-endotoxemia group compared with the endotoxemia group. CONCLUSIONS: Propofol attenuated cytokine responses, base deficit, and activation of neutrophils to endotoxemia. These findings suggest that propofol may inhibit inflammatory response and prevent the development of metabolic acidosis during endotoxemia. PMID- 10809291 TI - Effect of phased chest and abdominal compression-decompression cardiopulmonary resuscitation on myocardial and cerebral blood flow in pigs. AB - OBJECTIVE: This study was designed to assess the effects of a phased chest and abdominal compression-decompression cardiopulmonary resuscitation (CPR) device, Lifestick, vs. standard CPR on vital organ blood flow in a porcine CPR model. DESIGN: Prospective, randomized laboratory investigation using an established porcine model with instrumentation for measurement of hemodynamic variables, vital organ blood flow, blood gases, and return of spontaneous circulation. SETTING: University hospital research laboratory. SUBJECTS: Twelve domestic pigs. INTERVENTIONS: After 4 mins of untreated ventricular fibrillation, either the Lifestick CPR device (n = 6) or standard CPR (n = 6) was started and maintained for an additional interval of 6 mins before attempting defibrillation. MEASUREMENTS AND MAIN RESULTS: During CPR, but before epinephrine, use of the Lifestick CPR device resulted in significantly higher (p < .05) mean (+/- SD) coronary perfusion pressure (23+/-9 vs. 10+/-7 mm Hg), cerebral perfusion pressure (29+/-11 vs. 18+/-10 mm Hg), mean arterial pressure (49+/-10 vs. 36+/-13 mm Hg), end-tidal carbon dioxide (32+/-11 vs. 20+/-7 mm Hg), left ventricular myocardial blood flow (44+/-19 vs. 19+/-12 mL x min(-1) x 100 g(-1)), and total cerebral blood flow (29+/-10 vs. 14+/-12 mL x min(-1) x 100 g(-1)). After 45 microg/kg epinephrine, hemodynamic and vital organ blood flow variables increased to comparable levels in both groups. CONCLUSIONS: Compared with standard CPR, the Lifestick CPR device increased significantly hemodynamic variables and vital organ blood flow during CPR before epinephrine administration. PMID- 10809292 TI - Perfluorocarbon blocks tumor necrosis factor-alpha-induced interleukin-8 release from alveolar epithelial cells in vitro. AB - OBJECTIVE: To determine whether tumor necrosis factor (TNF)-alpha-induced interleukin (IL)-8 production by pulmonary alveolar epithelial cells is blocked by perfluorocarbon (PFC). DESIGN: Controlled, laboratory investigation of IL-8 production by pulmonary alveolar epithelial cells after exposure to PFC in vitro. SETTING: University research laboratory. SUBJECT: The human alveolar epithelial cell line with pulmonary type II (A549) cell properties. INTERVENTIONS: The A549 cells on a polycarbonate porous filter were stimulated either on the apical or the basolateral side with TNF-alpha. To determine TNF-alpha-induced IL-8 production, IL-8 was measured by using a human IL-8 kit in both control and experimental groups. MEASUREMENTS AND MAIN RESULTS: TNF-alpha stimulation induced a large increase in IL-8. When PFC was added to the medium immediately after TNF alpha stimulation, PFC separated the medium from the cells and IL-8 production was markedly reduced (TNF-alpha alone, 8342+/-470 pg vs. TNF-alpha followed by PFC, 417+/-88 pg, p < .05). Preincubation of A549 cells with PFC for 24 hrs before stimulation with TNF-alpha followed by removal of PFC did not affect IL-8 production (8834+/-204 vs. 8342+/-470 pg; p = NS). When added to the lower chamber, TNF-alpha also induced IL-8 production unaffected by the addition of PFC to the upper chamber. The decrease in TNF-alpha-induced IL-8 production depended on the time of PFC administration after the initiation of TNF-alpha stimulation. The earlier PFC was added, the more pronounced the diminution was in IL-8. CONCLUSIONS: PFC appears to function as a physical barrier, thus reducing cytokines produced by alveolar epithelial cells in vitro. This mechanism may partially explain the decreased inflammatory response observed during liquid ventilation in models of acute lung injury. PMID- 10809294 TI - Effect of mild hypothermia on cerebral oxygen uptake during gradual cerebral perfusion pressure decrease in piglets. AB - OBJECTIVE: To study the effect of mild hypothermia on cerebral oxygen metabolism and brain function in piglets during reduced cerebral blood flow because of gradual reduction of the effective cerebral perfusion pressure (CPP). DESIGN: Comparison of two randomized treatment groups: normothermic group (NT; n = 7) and hypothermic group (HT; n = 7). SETTING: Work was conducted in the research laboratory of the Institute for Pathophysiology, Friedrich Schiller University, Jena, Germany. SUBJECTS: Fourteen piglets (14 days old) of mixed German domestic breed. INTERVENTION: Animals were anesthetized and mechanically ventilated. An epidural balloon was gradually inflated to increase intracranial pressure to 25 mm Hg, 35 mm Hg, and 45 mm Hg every 30 mins at adjusted mean arterial blood pressures. After determination of baseline CPP (NT, 79+/-14 mm Hg; HT, 84+/-9 mm Hg), CPP was reduced to approximately 70%, 50%, and 30% of baseline (NT, 38.1+/ 0.5 degrees C; HT, 31.7+/-0.5 degrees C). MEASUREMENTS AND MAIN RESULTS: Every 25 mins after the gradual CPP reductions. Mild hypothermia induced a reduction of the cerebral metabolic rate of oxygen (CMRO2) to 50%+/-15% of baseline values (baseline values, 352+/-99 micromol x 100 g(-1) x min(-1)) (p < .05). Moreover, the electrocorticogram was altered to a pattern of reduced delta activity (p < .05) but unchanged higher frequency activity. The cerebral oxygen balance in HT animals remained improved until CPP reduction to 50%, indicated by a reduced cerebral arteriovenous difference of oxygen but elevated brain tissue Po2 (p < .05). Further CPP reduction gave rise to a strong CMRO2 reduction (NT, 19+/-21%; HT, 15+/-15%; p < .05). However, the high-frequency band of electrocorticogram was less reduced in hypothermic animals (p < .05). CONCLUSIONS: Mild whole body hypothermia improves cerebral oxygen balance by reduction of brain energy demand in juvenile piglets. The improvement of brain oxygen availability continues during a mild to moderate CPP decrease. A loss of the difference in CMRO2 between the hypothermic and normothermic piglets together with the fact that brain electrical activity was less suppressed under hypothermia during severe cerebral blood flow reduction indicates that hypothermic protection may involve some other mechanisms than reduction of brain oxidative metabolism. PMID- 10809293 TI - Bradykinin B2-receptor antagonism attenuates fatal cardiocirculatory breakdown induced by severe experimental pancreatitis. AB - OBJECTIVES: To investigate the impact of the long-acting bradykinin B2 receptor antagonist HOE 140 (Icatibant) on survival time in a model of severe porcine pancreatitis. DESIGN: Randomized, controlled intervention trial. SUBJECTS: Thirty domestic pigs of either gender anesthetized by intravenous application of piritramide, midazolam, and pancuronium and mechanically ventilated. INTERVENTIONS: Pancreatitis was induced by an injection of sodium taurocholate (5%, 1 mL/kg body weight [BW]) and enterokinase (10 U/kg BW). Control animals (group 1, n = 10) underwent the spontaneous course of the disease. In two treatment groups, Icatibant was administered either in a low (100 nmol/kg BW; group 2, n = 10) or in a high dosage (5000 nmol/kg BW; group 3, n = 10). MEASUREMENTS AND MAIN RESULTS: Mean survival time was significantly prolonged by Icatibant (controls, 6.6 hrs; group 2, 9.8 hrs; p = .022; group 3, 10.9 hrs; p = .007). Six hours postinduction, the decline of total peripheral resistance (52% of baseline) and cardiac index (92% of baseline) in controls was significantly improved by Icatibant, both in the low (16% and 44%; p < .05) and high (6% and 45%; p < .05) dosage. The concentrations of free, nonreceptor-bound kinin in plasma 6 hrs postinduction were significantly lower in controls than in groups 2 and 3 animals (111+/-50 vs. 208+/-40 and 237+/-52 fmol/mL, respectively). Six hours postinduction, the pretreatment with Icatibant was associated with significantly higher plasma concentrations of phospholipase A2 (controls, +1194%; group 2, +2000%; group 3, +2285% of baseline values) and interleukin-1 receptor antagonist (controls, 1900+/-800; group 2, 3100+/-800; group 3, 3600+/-800 pg/mL). In contrast, the increase of urinary trypsinogen activation peptides indicating local pancreatic damage (589+/-114 nmol/L in controls) was substantially attenuated by pretreatment with Icatibant (group 2, 467+/-102, NS; 352+/-91 nmol/L in group 3; p = .022 vs. controls). Systemic inflammatory reactions, however, as quantified by C-reactive protein and the extracellularly discharged neutrophil cytosolic inhibitor leukocyte neutral proteinase inhibitor were not influenced by the bradykinin B2-receptor antagonist. CONCLUSIONS: Pretreatment with the bradykinin B2 receptor antagonist Icatibant resulted in prolonged survival time and in delayed impairment of major macrocirculatory and pulmonary variables. Icatibant resulted in elevated concentrations of free, circulating kinin. This was associated with increased concentrations of phospholipase A2 and interleukin-1 receptor antagonist, suggesting that circulating kinins strengthen the activation of some mediator cascades, the association of which with the kinin metabolism requires further experimental clarification. Other variables indicating a systemic inflammatory response (C reactive protein, leukocyte neutral proteinase inhibitor) remained unaffected by Icatibant. Bradykinin antagonism distinctly ameliorated the local pancreatic damage, indicated by increased urinary concentrations of trypsinogen activation peptides. It is concluded that the kinin metabolism plays an important role in the pathophysiology of systemic complications after severe experimental pancreatitis. PMID- 10809295 TI - Prolonged hypernatremia controls elevated intracranial pressure in head-injured pediatric patients. AB - OBJECTIVE: To determine the effects continuous infusions of hypertonic saline (3% NaCl) on intracranial pressure (ICP) control and describe the physiologic effects of hypertonic saline administered to closed head injury children. DESIGN: Retrospective chart review. SETTINGS: Pediatric intensive care unit of a children's hospital. PATIENTS: Sixty-eight children with closed head injury. INTERVENTIONS: Intravenous infusion of 3% hypertonic saline to increase serum sodium to levels necessary to reduce ICP < or =20 mm Hg. MEASUREMENTS AND MAIN RESULTS: The patients enrolled had similar Injury Severity Scores. Treatment effectively lowered ICP in these patients and ICP was under good control a majority of the time. Only three patients (4%) died of uncontrolled elevation of ICP. No adverse effects of supraphysiologic hyperosmolarity such as renal failure, pulmonary edema, or central pontine demyelination, were noted. CONCLUSIONS: Hypertonic saline administration to children with closed head injury appears to be a promising therapy for control of cerebral edema. Further controlled trials are required to determine the optimal duration of treatment before widespread use is advocated. PMID- 10809296 TI - Use of hypertonic saline in the treatment of severe refractory posttraumatic intracranial hypertension in pediatric traumatic brain injury. AB - OBJECTIVES: To evaluate the effect of prolonged infusion of 3% hypertonic saline (514 mEq/L) and sustained hypernatremia on refractory intracranial hypertension in pediatric traumatic brain injury patients. DESIGN: A prospective study. SETTING: A 24-bed Pediatric Intensive Care Unit (Level III) at Children's Hospital. PATIENTS: We present ten children with increased intracranial pressure (ICP) resistant to conventional therapy (head elevation at 30 degrees, normothermia, sedation, paralysis and analgesia, osmolar therapy with mannitol, loop diuretic, external ventricular drainage in five patients), controlled hyperventilation (Pco2, 28-35 mm Hg), and barbiturate coma. We continuously monitored ICP, cerebral perfusion pressure (CPP), mean arterial pressure, central venous pressure, serum sodium concentrations, serum osmolarity, and serum creatinine. INTERVENTIONS: A continuous infusion of 3% saline on a sliding scale was used to achieve a target serum sodium level that would maintain ICP <20 mm Hg once the conventional therapy and barbiturate coma as outlined above failed to control intracranial hypertension. MEASUREMENTS AND MAIN RESULTS: The mean duration of treatment with 3% saline was 7.6 days (range, 4-18 days). The mean highest serum sodium was 170.7 mEq/L (range, 157-187 mEq/L). The mean highest serum osmolarity was 364.8 mosm/L (range, 330-431 mosm/L). The mean highest serum creatinine was 1.31 mg/dL (range, 0.4-5.0 mg/dL). There was a steady increase in serum sodium versus time zero that reached statistical significance at 24, 48, and 72 hrs (p < .01). There was a statistically significant decrease in ICP spike frequency at 6, 12, 24, 48, and 72 hrs (p < .01). There was a statistically significant increase in CPP versus time zero at 6, 12, 24, 48, and 72 hrs (p < .01). There was a statistically significant increase in serum osmolarity versus time zero at 12 hrs (p < .05) and at 24, 48, and 72 hrs (p < .01). Two patients developed acute renal failure and required continuous veno-venous hemodialysis; these were concurrent with an episode of sepsis and multisystem organ dysfunction. Both recovered full renal function with no electrolyte abnormalities at the time of discharge. CONCLUSION: An increase in serum sodium concentration significantly decreases ICP and increases CPP. Hypertonic saline is an effective agent to increase serum sodium concentrations. Sustained hypernatremia and hyperosmolarity are safely tolerated in pediatric patients with traumatic brain injury. Controlled trials are needed before recommendation of widespread use. PMID- 10809297 TI - Evaluation of ethical conflicts associated with randomized, controlled trials in critically ill children. AB - OBJECTIVE: To determine whether randomized, controlled trials (RCTS) of potentially life-sustaining therapies in critically ill infants and children cause an ethical conflict for physician investigators and if ethical conflicts affect protocol implementation. DESIGN: Descriptive survey. SUBJECTS: A convenience sample of 1,050 physicians from a national pediatric critical care meeting mailing list. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The survey return rate was 41% (n = 415). Of the returned surveys, 81% (n = 331) were answered by pediatric intensivists and fellows. The remaining 19% (n = 84) were completed by other physician subspecialists. Overall, 74% had experience with RCTs involving a potentially life-saving therapy (25% had experience with three or more trials, and 26% had never participated in this type of study). The vast majority of the respondents (96%) indicated that they believe RCTs of potentially life-sustaining therapies are ethical; however, only 10% stated that they never experienced an ethical conflict with these types of studies. Most respondents (84%) indicated that published data from uncontrolled trials may bias them toward an investigational therapy. Furthermore, only 35% of the respondents indicated that they always maintain strict protocol adherence when the condition of a control patient deteriorates and parents request the experimental treatment. There was a significant association between physicians who experienced an ethical conflict and the likelihood that they would do the following if the condition of a control patient deteriorated: fail to maintain strict protocol adherence (p = .05); alter the protocol in response to parental requests for the experimental treatment (p < .01); or seek compassionate use of the experimental treatment (p < .01). CONCLUSIONS: Although physicians consider RCTs of potentially life sustaining therapies ethical, they acknowledge that this type of study sometimes creates an ethical conflict. Published results of uncontrolled trials lead to investigator bias in randomized trials and preclude equipoise. Our results indicate that RCTs involving life-sustaining therapies may be biased, lack consistent protocol implementation, and raise concern that data from these studies are potentially flawed. PMID- 10809298 TI - Use of active noise cancellation devices in caregivers in the intensive care unit. AB - OBJECTIVE: Recent development of noise cancellation devices may offer relief from noise in the intensive care unit environment. This study was conducted to evaluate the effect of noise cancellation devices on subjective hearing assessment by caregivers in the intensive care units. DESIGN: Randomized, double blind. SETTING: Adult medical intensive care unit and pediatric intensive care unit of a teaching hospital. SUBJECTS: Caregivers of patients, including nurses, parents, respiratory therapists, and nursing assistants from a medical intensive care unit and pediatric intensive care, were enrolled in the study. INTERVENTION: Each participant was asked to wear the headphones, functional or nonfunctional noise cancellation devices, for a minimum of 30 mins. MEASUREMENTS: Subjective ambient noise level was assessed on a 10-point visual analog scale (VAS) before and during headphone use by each participant. Headphone comfort and the preference of the caregiver to wear the headphone were also evaluated on a 10 point VAS. Simultaneously, objective measurement of noise was done with a sound level meter using the decibel-A scale and at each of nine octave bands at each bedspace. RESULTS: The functional headphones significantly reduced the subjective assessment of noise by 2 (out of 10) VAS points (p < 0.05) in environments of equal objective noise profiles, based on decibel-A and octave band assessments. CONCLUSION: Noise cancellation devices improve subjective assessment of noise in caretakers. The benefit of these devices on hearing loss needs further evaluation in caregivers and critically ill patients. PMID- 10809299 TI - Amino acid loss and nitrogen balance in critically ill children with acute renal failure: a prospective comparison between classic hemofiltration and hemofiltration with dialysis. AB - HYPOTHESIS: Amino acid (AA) loss is not equivalent on continuous venovenous hemofiltration (CVVH) compared with continuous venovenous hemodiafiltration (CVVHD). Amino acid supplementation may be necessary to adjust for a greater clearance on CVVH to maintain nitrogen balance similar to that of CVVHD. OBJECTIVE: To compare AA losses and nitrogen balance between CVVH and CVVHD in children with acute renal failure. SETTING: Pediatric patients in the pediatric intensive care unit of a tertiary referral center. DESIGN: Prospective randomized crossover study in consecutive children who required hemofiltration. PATIENTS: A total of 12 plasma clearance studies for AA and urea, consisting of 24-hr collections of ultrafiltrate and urine for nitrogen balance, was performed on six patients during CVVH and CVVHD. Patients received total parenteral nutrition (TPN) with caloric intake 20% to 30% above their resting energy expenditure measured by indirect calorimetry and 1.5 g/kg/day protein of TPN. Study conditions were comprised of 2 L/hr/1.73 m2 of dialysate or prefiltered replacement fluid and hemofilter flow rates of 4 mL/kg/min were maintained for all patients. METHODS AND MAIN RESULTS: Amino acid clearances were greater on CVVH than CWHD, except for glutamic acid, where clearance was 6.73+/-2.31 (SEM) mL/min/1.73 m2 on CVVH and 7.59+/-2.79 mL/min/1.73 m2 for CVVHD (NS). The clearance difference between the two modalities was 30%. Urea clearance was equivalent (30.1+/-1.74 mL/min/1.73 m2 and 29.0+/-.97 mL/min/1.73 m2) for CVVH and CVVHD, respectively. Amino acid loss on CVVH and CVVHD was similar (12.50+/ 1.29 g/day/1.73 m2 vs. 11.61+/-1.86 g/day/1.73 m2, respectively), representing 12% and 11%, respectively, of the daily protein intake. The catabolic state, as measured by urea nitrogen appearance, was high for all patients during the 48-hr study period with a mean of 291 mg/kg/day during CVVH, and 245 mg/kg/day for CVVHD. Nitrogen balance varied from a negative 12.95 g/day/1.73 m2 to a positive 4.93 g/day/1.73 m2 on CVVH and a negative 7.69 g/day/1.73 m2 to a positive 5.50 g/day/1.73 m2 on CVVHD. CONCLUSIONS: Clearance of AA is greater on CVVH than on CVVHD, but no significant difference in AA loss was present between the two therapies. Nitrogen balance often is not met on either therapy when a standard 1.5 g/kg/day protein and a resting energy expenditure of 120% to 130% of calories is delivered by TPN. PMID- 10809300 TI - Energy expenditure in critically ill children. AB - OBJECTIVES: To measure energy expenditure in critically ill children and compare it with the energy expenditure predicted by recommended formulas, and relate the measured energy expenditure to nutritional and clinical indices. DESIGN: A prospective, clinical study. SETTING: Tertiary care pediatric intensive care unit in a university children's hospital. PATIENTS: A total of 37 patients with critical illness who were mechanically ventilated for > or =24 hrs were studied. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Chronic protein-energy malnutrition (CPEM) and acute protein-energy malnutrition were defined by the Waterlow's stages and fat and protein stores were classified as defined by Frisancho, Ryan, and Martinez. Severity of illness was assessed by the Pediatric Risk of Mortality Score, the Therapeutic Intervention Scoring System, and indices of organ failure. Oxygen consumption, carbon dioxide production, and the respiratory quotient were measured by indirect calorimetry, and energy expenditure (MEE) was calculated using the modified Weir formula. Resting energy expenditure (PBMR), predicted energy expenditure, and caloric intake were calculated using recommended formulas. A total of 77 measurements were made in 37 children. MEE was significantly lower than PBMR as estimated by all equations except the Talbot equations. MEE was significantly lower than predicted energy expenditure and the recommended daily allowances. On the first day, the MEE/PBMR ratio was <0.9 in 56.8%, 0.9-1.1 in 21.6%, and >1.1 in 21.6% of patients. MEE did not differ significantly among disease groups or between medical and surgical patients. There was no difference in MEE with or without neuromuscular blockade. MEE was lower in the presence of multiple organ system failure (MOSF) (1019 + 166 kcal/m2 without MOSF vs. 862 + 241 with MOSF; p = .025). A total of 21% had CPEM and 8.1% had acute protein-energy malnutrition. Multivariate stepwise regression analysis showed that the protein intake, midarm muscle area, midarm fat area, the use of vasoactive agents, and sedation correlated with MEE (p < .05). With CPEM, MEE was correlated to the severity of illness (p < .05). Patients at risk for protein stores depletion (midarm muscle areas 1 and 2) had a higher incidence of MOSF compared with nutritionally normal children (p < .05), whereas patients with fat stores depletion (midarm fat area 2) had a higher probability of death (50% vs. 6%, respectively). CONCLUSIONS: Recommended daily allowances and energy expenditure predicted by using a stress-related correction to the resting energy expenditure grossly overestimate MEE. MEE is close to PBMR and in many patients, it is lower than PBMR. MEE that is lower than PBMR is associated with a higher morbidity. Nutritional repletion should thus be based on MEE to avoid the problems of over- or underfeeding. PMID- 10809301 TI - Relationship of illness severity and length of stay to functional outcomes in the pediatric intensive care unit: a multi-institutional study. AB - OBJECTIVE: The purpose of this study was to establish relationships between illness severity, length of stay, and functional outcomes in the pediatric intensive care unit (PICU) by using multi-institutional data. We hypothesized that a positive relationship exists between functional outcome scores, severity of illness, and length of stay. DESIGN: The study used a prospective multicentered inception cohort design. SETTING: The study was conducted in 16 PICUs across the United States that were member institutions of the Pediatric Critical Care Study Group of the Society of Critical Care Medicine. PATIENTS: In total, 11,106 patients were assessed, representing all admissions to these intensive care units for 12 consecutive months. MEASUREMENTS: Functional outcomes were measured by the Pediatric Overall Performance Category (POPC) and Pediatric Cerebral Performance Category (PCPC) scales. Both scales were assessed at baseline and discharge from the PICU. Delta scores were formed by subtracting baseline scores from discharge scores. Other measurements included admission Pediatric Risk of Mortality scores, age, operative status, length of stay in the PICU, and diagnoses. Interrater reliability was assessed by using a set of ten standardized cases on two occasions 6 months apart. MAIN RESULTS: Baseline, discharge, and delta POPC and PCPC outcome scores were associated with length of stay in the PICU and with predicted risk of mortality (p < .01). Incorporation of baseline functional status in multivariate length of stay analyses improved measured fit. Mild baseline cerebral deficits in children were associated with 18% longer PICU stays after controlling for other patient and institutional characteristics. Moderate and severe baseline deficits for both the POPC and PCPC score predict increased length of stay of between 30% and 40%. On the standardized cases, interrater consensus was achieved on 82% of scores with agreement to within one neighboring class for 99.7% of scores. CONCLUSIONS: These data establish current relationships for the POPC and PCPC outcome scales based on multi-institutional data. The reported relationships can be used as reference values for evaluating clinical programs or for clinical outcomes research. PMID- 10809302 TI - Management and outcomes of delayed sternal closure after cardiac surgery in neonates and infants. AB - OBJECTIVES: To investigate the efficacy, safety, and patterns of management of open sternotomy and delayed sternal closure in infants who were left with an open sternum after cardiac surgery and to assess these patterns for possible correlation with outcome. DESIGN: Retrospective chart review with statistical analysis. SETTING: Pediatric cardiac surgery service at a regional referral center based in an urban university teaching hospital. PATIENTS: All 128 patients <1 yr of age who were left with an open sternum after cardiac surgery with cardiopulmonary bypass during the 4-yr period from July, 1992 to June, 1996. INTERVENTIONS: Procedures for managing open sternotomy and delayed sternal closure were analyzed retrospectively. No interventions were undertaken for the study. MEASUREMENTS AND RESULTS: Of the 128 patients, 14 (11%) died before sternal closure; delayed sternal closure was performed in the remaining 114. Of these 114, 13 died in the early postoperative period. During sternal closure, significant increases were noted in pulmonary arterial (from 21.1+/-7.6 mm Hg to 26.1+/-6.5 mm Hg; p = .006), left atrial (from 8.4+/-3.4 mm Hg to 11.5+/-3.7 mm Hg; p < .001), and right atrial pressures (from 7.3+/-2.5 mm Hg to 9.8+/-2.5 mm Hg; p < .001). In addition, mean airway pressure (from 7.4+/-2.0 mm Hg to 8.6+/ 2.4 mm Hg; p < .001) and peak inspiratory pressure (from 29.3+/-5.4 mm Hg to 31.3+/-5.6 mm Hg; p = .004) increased. Sternal wound infection occurred in one patient. CONCLUSIONS: Delayed sternal closure is an effective approach to the management of neonates and infants at risk for hemodynamic, respiratory, or hemostatic instability early after cardiac surgery. Significant changes in hemodynamics and respiratory variables occur during sternal closure, often requiring adjustment of inotropic and ventilatory management. (Crit Care Med 2000; 28: 1180-1184) PMID- 10809303 TI - Clinical evaluation of a new, point-of-care hemocytometer. AB - OBJECTIVE: This study was designed to compare results obtained with a new point of-care hemocytometer with those of two established (point-of-care and laboratory based) instruments. DESIGN: To compare CBC values between established laboratory based and point-of-care instruments, measurements were performed on routinely obtained blood specimens for CBC analysis in our institutional laboratory (phase I) and on specimens from cardiac surgical patients before initiation of cardiopulmonary bypass and after discontinuation of cardiopulmonary bypass in phase II. SETTING: Surgical and hospitalized patients at a tertiary care center. PATIENTS: Measurements were obtained by using blood specimens obtained from 141 hospitalized patients from different services (phase I) or from a consecutive series of 204 patients undergoing cardiac operations (phase II). MEASUREMENTS AND MAIN RESULTS: Hemoglobin (HGB), platelet count (PLT), red blood cell count, and white blood cell count (WBC) were measured with two on-site and one laboratory based instruments. Hematocrit (HCT) was calculated by using measured variables. Linear regression demonstrated good correlations between Ichor and T540 HGB (r2 = .95), HCT (r2 = .95), PLT (r2 = .94), and WBC (r2 = .95) results (n = 408); similarly, good correlations were observed with Coulter STKS HGB (r2 = .92), HCT (r2 = .91), and PLT (r2 = .94) results (n = 141). The relationship between Ichor and Coulter STKS WBC (r2 = .27) was poor; however, when two Ichor-derived outlier values (>50) were excluded, the relationship was very good (r2 = .99). Bias analysis (mean +/- SD) demonstrated similar results between Ichor and T540 HGB (0.003+/-0.5), HCT (-0.21+/-1.5), WBC (0.79+/-1.3), and PLT values (-9.2+/-16.6) as well as STKS HGB (-0.08+/-0.7), HCT (-0.69+/-2.3), WBC (-0.62+/-5.8), and PLT values (-10.2+/-21.4). CONCLUSIONS: The Ichor hemocytometer provides accurate hematologic results that can facilitate rapid quantitative assessment of CBC variables and thus may be clinically useful, especially in critically ill patients. PMID- 10809304 TI - The first decade of the American Board of Internal Medicine certification in critical care medicine: an overview of examinees and certificate holders from 1987 through 1996. AB - OVERVIEW: This study reviews the first decade of critical care medicine (CCM) certification by the American Board of Internal Medicine (1987-1996). Included are the characteristics of examinee and certificate-holder groups; examination performances from different underlying disciplines of internal medicine, with or without formal CCM training; and the influence of background and a training program as correlates of examination performance. DATA SOURCES: The CCM certification examination has been offered biennially since November 1987. Performance data on the American Board of Internal Medicine examinations in internal medicine and its subspecialties and added qualifications were available for candidates taking the CCM examinations. For examinees with formal CCM training, residency program director ratings, and information regarding the program characteristics of size and percentage of United States and Canadian medical graduates were also available. STUDY SELECTION: All examinees who ever attempted certification were included in this study. The study cohort for each of the five examination administrations consists of all first-time takers. CONCLUSIONS: Cohort sizes have decreased since formal training became an admission requirement in 1993. Percentages of International Medical Graduates and women attempting and achieving certification have increased steadily. Examination performance was positively associated with formal training, internal medicine examination performance, recent medical training, and pulmonary disease certification. For those with formal training, performance was also positively associated with training program director ratings of overall clinical competence and completion of a training program with a higher proportion of United States and Canadian medical graduates. PMID- 10809305 TI - Bereavement follow-up after critical illness. AB - OBJECTIVE: To describe the establishment and initial activity of a Bereavement Follow-up Service for next-of-kin of patients who died in an intensive care unit (ICU) and to quantify aspects of their experience including quality of intensive care service and the early impact on next-of-kin of their bereavement. DESIGN: Cross-sectional prospective study, which was conducted by a structured telephone interview. SETTING: A 14-bed adult general ICU in a tertiary university hospital. SUBJECTS: A total of 99 next-of-kin of patients who died in ICU. INTERVENTIONS: Referral to other agencies if requested. MEASUREMENTS AND MAIN RESULTS: Attempts were made to contact the next-of-kin of all 151 patients who died in 1995, and 104 were contacted. Five declined to be interviewed. The results refer to 99 who consented to telephone interview a median of 33 days after the death. A total of 84 considered themselves well informed during the intensive care period, 76 understood the fatal sequence of events but 19 of them would have liked more information. A total of 77 had positive comments about the quality of care, most commonly about compassionate behavior (58), but 30 had negative comments, most commonly about poor communication (13). Only 7 were living alone, 85 had resumed normal activities, 40 of 47 workers had returned to work, 58 had sleep disturbance at some stage (still present in 44), but only 12 were taking sedatives or antidepressants. A total of 32 had financial difficulties and 21 were referred to other agencies, most commonly grief counselors. CONCLUSIONS: We were disappointed to contact only two thirds of next-of-kin, but results from these subjects demonstrated a high level of satisfaction with the care given. Nevertheless, some were dissatisfied with the quality of service they experienced. Most had resumed their normal activities, including work, and few were living alone. However, sleep disturbance and financial difficulty were common, and some requested help from other support agencies. PMID- 10809306 TI - Use of intensive care-specific interventions in major teaching and other hospitals: a regional comparison. AB - OBJECTIVE: To compare the use of 40 specific medical interventions in intensive care units (ICUs) of major teaching and other hospitals DESIGN: Retrospective cohort study. SETTING: Thirty-eight ICUs in 28 hospitals in a large metropolitan region. PATIENTS: A total of 12,929 consecutive eligible admissions to medical, surgical, neurologic, or mixed medical/surgical ICUs between January 1, and June 30, 1994. MEASUREMENTS: The use of 40 diagnostic and therapeutic interventions during the first 24 hrs of ICU admission were obtained from patient medical records and a weighted intervention score was determined for each patient. Admission severity of illness was measured by using the Acute Physiology and Chronic Health Evaluation III methodology. MAIN RESULTS: Patients at the five teaching hospitals had a greater severity of illness (mean predicted risk of in hospital death, 15.1%+/-21.9% vs. 11.2%+/-19.0%; p < .01) than patients at the 23 other hospitals. Patients at major teaching hospitals also had higher mean intervention scores (3.5+/-4.9 vs. 2.3+/-3.7; p < .01). Differences in intervention scores persisted after controlling for severity of illness, admission diagnosis, and admission source. However, scores varied among the major teaching hospitals. When examined individually, only three of the five major teaching hospitals had higher (p < .05) interventions scores, compared with other hospitals, whereas one had a lower (p < .05) intervention score. CONCLUSIONS: Patients in ICUs at major teaching hospitals were, in aggregate, more likely to receive diagnostic and therapeutic interventions than patients at other hospitals. Variation among major teaching hospitals suggests that factors other than teaching status also affect the use of these interventions. PMID- 10809307 TI - Chylothorax: a novel therapy. AB - OBJECTIVE: To report a novel therapy for chylothorax. DESIGN: Case report. PATIENT: Severely ill neonate with congestive heart failure and high surgical risk. INTERVENTIONS: Standard therapy for chylous effusion with the addition of pressure control ventilation and positive end-expiratory pressure (high mean airway pressure). RESULTS: Rapid resolution of effusion with elimination of excessive fluid needs and correction of coagulation profile. CONCLUSION: Increasing mean airway pressure by pressure control ventilation and positive end expiratory pressure should be considered as a therapeutic intervention in patients with persistent chylous effusions. PMID- 10809308 TI - Use of recruitment maneuvers and high-positive end-expiratory pressure in a patient with acute respiratory distress syndrome. AB - OBJECTIVE: To present the use of a novel high-pressure recruitment maneuver followed by high levels of positive end-expiratory pressure in a patient with the acute respiratory distress syndrome (ARDS). DESIGN: Observations in one patient. SETTING: The medical intensive care unit at a tertiary care university teaching hospital. PATIENT: A 32-yr-old woman with severe ARDS secondary to streptococcal sepsis. INTERVENTIONS: The patient had severe gas exchange abnormalities because of acute lung injury and marked lung collapse. Attempts to optimize recruitment based on the inflation pressure-volume (PV) curve were not sufficient to avoid dependent lung collapse. We used a recruitment maneuver using 40 cm H2O of positive end-expiratory pressure (PEEP) and 20 cm H2O of pressure controlled ventilation above PEEP for 2 mins to successfully recruit the lung. The recruitment was maintained with 25 cm H2O of PEEP, which was much higher than the PEEP predicted by the lower inflection point (P(Flex)) of the PV curve. MEASUREMENTS AND MAIN RESULTS: Recruitment was assessed by improvements in oxygenation and by computed tomography of the chest. With the recruitment maneuvers, the patient had a dramatic improvement in gas exchange and we were able to demonstrate nearly complete recruitment of the lung by computed tomography. A PV curve was measured that demonstrated a P(Flex) of 16-18 cm H2O. CONCLUSION: Accumulating data suggest that the maximization and maintenance of lung recruitment may reduce lung parenchymal injury from positive pressure ventilation in ARDS. We demonstrate that in this case PEEP alone was not adequate to recruit the injured lung and that a high-pressure recruitment maneuver was required. After recruitment, high-level PEEP was needed to prevent derecruitment and this level of PEEP was not adequately predicted by the P(Flex) of the PV curve. PMID- 10809309 TI - Favorable outcome in a large left heart air embolism: lessons from an unusual complication of a noninvasive chest scan. AB - OBJECTIVE: To report an unusual life-threatening complication of the performance of a computed tomographic (CT) scan of the chest. DESIGN: Case report. SETTING: University hospital. PATIENT: An intubated patient with blunt thoracic trauma. INTERVENTION: Performance of a CT scan of the chest at full inspiration. MAIN RESULT: With air insufflation, a large left ventricular air embolism occurred as a consequence of an airway breach, revealed by the simultaneous existence of a mild bilateral anterior pneumothorax. CONCLUSION: CT scan of the chest in patients at risk of airway breach (patients with acute respiratory distress syndrome, trauma patients) should first be performed at full expiration, not full inspiration. PMID- 10809310 TI - Controversy: is there a "renal dose" dopamine? PMID- 10809311 TI - Interleukin-8, neuroinflammation, and secondary brain injury. PMID- 10809312 TI - Blood pressure support in patients with multiple organ failure: is more better. PMID- 10809313 TI - Procalcitonin: clinical tool or laboratory curiosity? PMID- 10809314 TI - Procalcitonin: THE marker of sepsis? PMID- 10809315 TI - Toward a better outcome from symptomatic vasospasm. PMID- 10809316 TI - Homocysteine and critical illness. PMID- 10809317 TI - Evaluation of picture archiving technology. PMID- 10809318 TI - Respiratory failure in bone marrow transplant patients. PMID- 10809319 TI - Chest radiograph or computed tomography in the intensive care unit? PMID- 10809320 TI - Inhaled nitric oxide: more than a selective pulmonary vasodilator. PMID- 10809321 TI - Pressors for cardiopulmonary resuscitation: is there a new kid on the block? PMID- 10809322 TI - Goldilocks and the three cytokines. PMID- 10809323 TI - A new phased chest and abdominal compression-decompression cardiopulmonary device holds promise for use in humans. PMID- 10809324 TI - The anti-inflammatory effects of perfluorocarbons: let's get physical. PMID- 10809325 TI - Is reduced cerebral perfusion pressure better tolerated during hypothermia? PMID- 10809326 TI - Should we be using hypertonic saline to treat intracranial hypertension? PMID- 10809327 TI - Are randomized controlled trials ethical in critically ill children? PMID- 10809328 TI - The use and interpretation of indirect calorimetry in critically ill patients. PMID- 10809329 TI - Review of the rationale for delayed sternal closure. PMID- 10809330 TI - Internal medicine based critical care training: straight internal medicine versus pulmonary. PMID- 10809331 TI - Bereavement follow-up to families after death in the intensive care unit. PMID- 10809332 TI - Inducible nitric oxide synthase inhibition in sepsis: what is gained? PMID- 10809333 TI - Probiotic containing fermented milk supplement may improve the institution of early enteral nutrition. PMID- 10809334 TI - Better equations and formulas. PMID- 10809335 TI - Apnea threshold and pediatric brain death. PMID- 10809336 TI - The supranormal oxygen delivery trials controversy. Dobutamine in Sepsis Study Group. PMID- 10809337 TI - Understanding the management of heart failure. PMID- 10809338 TI - HFSA guidelines for management of patients with heart failure caused by left ventricular systolic dysfunction--pharmacological approaches. Heart Failure Society of America. PMID- 10809339 TI - Gender-based differences and menstrual cycle-related changes in specific diseases: implications for pharmacotherapy. AB - Pharmacists should be aware of gender-based differences and menstrual cycle related changes in six diseases: asthma, arthritis, migraine, diabetes, depression, and epilepsy. In general, women report symptoms of physical illness at higher rates, visit physicians more frequently, and make greater use of other health care services than men. Whereas reasons for these gender differences are not fully clear, a combination of biologic, physiologic, social, behavioral, psychologic, and cultural factors most likely contributes. A significant percentage of women with asthma, arthritis, migraine, diabetes, depression, or epilepsy experience worsening of their disease premenstrually. The mechanism is unknown, but is speculated to be multifactorial because of many endogenous and exogenous modulators and mediators of each disease. As part of general therapy for cycle-related exacerbations of any one of these disorders, patients should be encouraged to use a menstrual calendar to track signs and symptoms for two to three cycles; if cyclic trends are identified, the women should anticipate exacerbations and avoid triggering factors. Cyclic modulation with pharmacotherapy may be attempted. If unsuccessful, a trial of medical ovulation suppression with a gonadotropin-releasing hormone (GnRH) analog may be warranted. If that is successful, continuous therapy with a GnRH analog and steroid add-back therapy or less expensive alternatives may be effective. If pharmacotherapy is impractical, hysterectomy and bilateral oophorectomy with estrogen replacement therapy is a last resort. Gender differences and menstrual cycle-related changes are important areas for clinical and mechanistic research. PMID- 10809340 TI - Contrast medium-induced nephrotoxicity: pathophysiology and prevention. AB - Contrast medium-induced nephrotoxicity (CMN) is a common form of iatrogenic acute renal failure. Typically, patients experience changes in serum creatinine or creatinine clearance between 1 and 5 days after exposure to a contrast medium, but they rarely require dialysis. The mechanism for CMN is not understood, but renal insufficiency, dehydration, and congestive heart failure are risk factors. The frequency of CMN with high-osmolality versus low-osmolality media is controversial. Prophylaxis can reduce CMN. Of many different strategies, hydration with normal saline before and after exposure offers the best protection with the fewest adverse effects. PMID- 10809341 TI - The effect of ritonavir on the pharmacokinetics of meperidine and normeperidine. AB - STUDY OBJECTIVE: To determine the effects of ritonavir on the pharmacokinetics of meperidine and normeperidine. DESIGN: Open-label, crossover, pharmacokinetic study. SETTING: United States government research hospital. SUBJECTS: Eight healthy volunteers who tested negative for the human immunodeficiency virus. INTERVENTION: Subjects received oral meperidine 50 mg and had serial blood samples collected for 48 hours. They then received ritonavir 500 mg twice/day for 10 days, followed by administration of a second 50-mg meperidine dose and collection of serial samples. MEASUREMENTS AND MAIN RESULTS: Plasma samples were assayed for meperidine, normeperidine, and ritonavir. Meperidine's area under the curve (AUC) decreased in all subjects by a mean of 67+/-4% in the presence of ritonavir (p<0.005). Mean +/- SD maximum concentration was decreased from 126+/ 47 to 51+/-21 ng/ml. Normeperidine's mean AUC was increased 47%, suggesting induction of hepatic metabolism. CONCLUSION: Meperidine's AUC is significantly reduced, not increased, by concomitant ritonavir. Based on these findings, the risk of narcotic-related adverse effects from this combination appears to be minimal. However, increased concentrations of normeperidine suggest a potential for toxicity with increased dosages or long-term therapy. PMID- 10809342 TI - Pharmacokinetics and pharmacodynamics of ceftizoxime in patients with dosages adjusted for renal function. AB - STUDY OBJECTIVE: Our institution developed dosing guidelines for patients with renal impairment based on pharmacokinetic data and class-specific pharmacodynamics. Ceftizoxime was chosen as a model agent to evaluate if the modified guidelines achieved similar minimal plasma concentration (Cp(min)) and time above the minimum inhibitory concentration of the infecting organism (T>MIC) in patients with renal impairment versus those with normal renal function. DESIGN: Prospective pharmacokinetic and pharmacodynamic evaluation of ceftizoxime dosages. SETTING: University-affiliated hospital. PATIENTS: Forty-three patients with suspected or documented infection were enrolled and classified into four groups based on creatinine clearance (Cl(cr); ml/min): group 1, above 100; group 2, 61-99; group 3, 31-60; and group 4, 15-30. INTERVENTIONS: Ceftizoxime serum concentrations were obtained at steady state. MEASUREMENTS AND MAIN RESULTS: Pharmacokinetic and pharmacodynamic parameters were calculated. As expected, clearance and elimination rate constant were reduced, and half-life tended to be greater in patients with renal impairment. The Cp(min) and area under the concentration-time curve over 24 hours were similar between groups (p=0.39, p=0.42). The T>MIC was 100% for all patient isolates, and 90% or more versus our clinical strain for all groups. Clinical outcomes were similar among all groups. CONCLUSION: Our dosing guidelines achieved similar Cp(min) among all groups of patients. Our results support that recommendations for dosing adjustments should be based on pharmacokinetic data and must also consider pharmacodynamic parameters. PMID- 10809343 TI - Absorption of phenytoin from rectal suppositories formulated with a polyethylene glycol base. AB - STUDY OBJECTIVE: To compare phenytoin pharmacokinetics following administration of an oral suspension and a rectal suppository formulated with a polyethylene glycol base. DESIGN: Unblinded, single-dose, randomized, crossover trial. SETTING: University-affiliated pharmacokinetics and biopharmaceutics laboratory. SUBJECTS: Six healthy subjects. INTERVENTION: Subjects were given a single 200-mg dose of phenytoin as an oral suspension and a rectal suppository separated by a 1 week washout. MEASUREMENTS AND MAIN RESULTS: Blood for plasma phenytoin concentrations was obtained at baseline and 0.5, 1, 2, 4, 6, 8, 10, 12, and 24 hours after administration. Plasma was analyzed by high-performance liquid chromatography (coefficient of variation < 6%) for total phenytoin concentration. Phenytoin maximum concentration (Cmax), time to Cmax (Tmax), time to first measurable concentration (Tlag), and area under the curve from time zero to time of last measurable concentration (AUClast) were estimated for oral and rectal administration by WinNonlin (v 1.1) and compared using Wilcoxon's signed rank test (p<0.05 for statistical significance). Two subjects did not have detectable plasma phenytoin concentrations after rectal administration. For the other four subjects, median rectal Cmax was significantly lower than oral Cmax (0.4 vs 1.9 microg/ml, p=0.028), median rectal Tmax did not differ from oral Tmax (11.9 vs 8.0 hrs, p=0.465), and median rectal AUClast, although highly variable, was significantly lower than oral AUClast (5.4 vs 36.2 microg x hr/ml, p=0.046). No Tlag was seen after oral administration, but with rectal administration the median Tlag was 2 hours. The estimated relative bioavailability of rectal phenytoin suppositories based on AUC0-24 was 4.7%, with individual values ranging from 0-58.3%. CONCLUSION: It appears that absorption of phenytoin from polyethylene glycol rectal suppositories in healthy subjects is highly variable and unpredictable. Thus this formulation is not recommended. PMID- 10809344 TI - Consumer use of St. John's wort: a survey on effectiveness, safety, and tolerability. AB - Despite its poorly described pharmacology, effectiveness, and safety, use of St. John's wort (SJW) is largely unsupervised and unexplored, and can potentially lead to adverse outcomes. We conducted a telephone survey of 43 subjects who had taken SJW to assess demographics, psychiatric and medical conditions, dosage, duration of use, reason for use, side effects, concomitant drugs, professional consultation, effectiveness, relapse, and withdrawal effects. Most subjects reported taking SJW for depression, and 74% did not seek medical advice. Mean dosage was 475.6+/-360 mg/day (range 300-1200 mg/day) and mean duration of therapy was 7.3+/-10.1 weeks (range 1 day-5 yrs). Among 36 (84%) reporting improvement, 18 (50%) had a psychiatric diagnosis. Twenty (47%) reported side effects, resulting in discontinuation in five (12%) and one emergency room visit. Two consumers experienced symptoms of serotonin syndrome and three reported food drug interactions. Thirteen consumers experienced withdrawal symptoms and two had a depressive relapse. These data suggest the need for greater consumer and provider awareness of the potential risks of SJW in self-care of depression and related syndromes. PMID- 10809345 TI - Inappropriate drug use among community-dwelling elderly. AB - This study examined inappropriate drug use defined by updated criteria among respondents in the second and third in-person waves of the Duke Established Populations for Epidemiologic Studies of the Elderly. Information about sociodemographics, health status, access to health care, and drug use was determined by in-home interviews. Drug use was coded for therapeutic class and appropriateness by applying explicit criteria. Among participants, 27% of the second and 22.5% of the third in-person wave took one or more inappropriate agents. Of these drugs, the most common therapeutic classes were central nervous system and cardiovascular. Longitudinal multivariate analyses found that persons taking several prescription drugs, those having continuity of care, those who previously took inappropriate drugs, and those with many health visits were most likely (p<0.05) to use inappropriate drugs. We conclude that inappropriate drug use is common among community-dwelling elderly. PMID- 10809346 TI - Pharmacist-designed and -implemented pharmaceutical care plan for antipsychotic induced movement disorders. AB - We developed a systematic approach to assess the presence, severity, and management of extrapyramidal symptoms (EPS) in patients treated with antipsychotics. Patients were evaluated by the Modified Simpson-Angus scale, Abnormal Involuntary Movement Scale, and Dyskinesia Identification System: Condensed User Scale. We completed 235 sets of evaluations in 83 patients. A pharmaceutical intervention was proposed in 54% (130) of evaluations, of which 82% (107) were accepted and followed. In 93% (99) evaluations in which a recommendation was followed, clinical outcome was positive. The most common intervention was reducing the dosage or discontinuing the antidyskinetic agent, most often an anticholinergic (55% of cases). Our results show that detailed monitoring of EPS in a clinical pharmacist-operated clinic promotes rational drug therapy, limits unnecessary drugs, and improves clinical outcome of patients with EPS. PMID- 10809347 TI - Response of complicated methicillin-resistant Staphylococcus aureus endocarditis to the addition of trovafloxacin. AB - The newer fluoroquinolones have many properties such as safety, bioavailability, and tissue penetration that make them attractive in the therapy of complicated infections. Unfortunately, the rapid development of resistance by Staphylococcus aureus to ciprofloxacin has dampened interest in these agents for serious staphylococcal infections. A patient with right-sided methicillin-resistant Staphylococcus aureus (MRSA) endocarditis with a complicated clinical course received trovafloxacin in addition to vancomycin and rifampin. He was initially treated with vancomycin, gentamicin, and rifampin for serious MRSA infection, but because of complications, including septic central nervous system emboli, persistent fever, and leukocytosis, gentamicin was stopped and trovafloxacin begun. After this addition the patient improved and completely recovered. In vitro and animal model data show that many newer fluoroquinolones have excellent activity against S. aureus, including MRSA, and are also less likely to induce resistance. Animal models of endocarditis support their efficacy in serious staphylococcal infections. PMID- 10809348 TI - Waste generation of drug product samples versus prescriptions obtained through pharmacy dispensing. AB - Drug samples are often packaged differently from bulk packaging and thus they may contain a disproportionate amount of waste material. Fifteen drug samples were obtained from seven pharmaceutical companies and the packaging materials were weighed after the samples were removed. The waste produced by the samples was determined for a standard amount of drug and compared with the weight of the waste produced when the same quantity of drug was dispensed through a pharmacy. The average weight of the sample package for a standard course of therapy was significantly greater (p< or =0.05) than that of the pharmacy-dispensed prescription waste weight. The former was 5+/-4.5-fold heavier than the latter. The waste generated by drug samples in the United States was determined to be 5740 metric tons/year. PMID- 10809349 TI - Comparison of five beta-lactam antibiotics against common nosocomial pathogens using the time above MIC at different creatinine clearances--a comment. PMID- 10809350 TI - The clinical pharmacist as principal investigator: a commentary from the American College of Clinical Pharmacy. PMID- 10809351 TI - Cellular compartmentation of lysozyme and alpha-amylase in the mouse salivary glands. Immunogold approaches at light and electron microscopy level. AB - The research was planned to study the subcellular distribution of enzymatic secretory products within the secretory structures of the mouse major salivary glands at light and electron microscopy level by immunogold silver stain (IGSS) technique and double-sided post-embedding immunogold binding and silver amplification in order to speculate about their compartmentation. In particular, we experimented the above immunogold labeling approaches to localize the lysozyme and to verify its distribution patterns in relation to another secretion enzyme, alpha-amylase. Co-presence of lysozyme and alpha-amylase was observed in the convoluted granular tubule cells of the submandibular gland and in the demilunar cells of the sublingual gland as well as in the electron-dense regions of the mottled secretory granules in the parotid gland. Exclusive binding patterns of lysozyme were observed in the acinar cells of the submandibular and sublingual glands where alpha-amylase did not occur. PMID- 10809352 TI - Duodenal endocrine cells in mice with particular regard to age-induced changes. AB - Duodenal endocrine cell types in four age groups of NMRI mice (1, 3, 12 and 24 months old) were identified by immunocytochemistry and quantified by computerized image analysis. Whereas the number of secretin-immunoreactive cells was significantly increased in the 24-month-old group, the number of GIP immunoreactive cells was reduced in 12-month-old compared with 3-month-old mice. The number of somatostatin-immunoreactive cells was fewer in both the 12- and 24 month-olds vis-a-vis the 3-month-old mice. Whereas serotonin-immunoreactive cells were fewer in both 1-month-old and 12-month-old mice, they were more numerous in 24-month-old mice then in the 3-month-old ones. The number of gastrin/CCK immunoreactive cells was unaffected by age. The cell secretory index (CSI) of secretin- and serotonin-immunoreactive cells was increased in the 24-month-old mice vis-a-vis the 3-month-old ones and the CSI of GIP- and somatostatin immunoreactive cells was increased in 12-month-old mice vis-a-vis 3-month-old mice. In contrast, the CSI of somatostatin- and serotonin-immunoreactive cells in 1-month-old mice was lower than that of 3-month-old-mice. The nuclear volume of secretin-, GIP-, gastrin/CCK- and serotonin-immunoreactive cells was less in 1 month-olds than in 3-month-old mice. Whereas the nuclear volume of somatostatin immunoreactive cells was decreased in 12-month-old animals, that of gastrin/CCK- and serotonin-immunoreactive cells was greater in 24-month-old mice than in 3 month-old ones. It is concluded that these changes may be secondary to structural and functional changes in the gastrointestinal tract caused by ageing. It is possible that these changes are involved in the development of dysfunction of the gut observed at advanced age. PMID- 10809353 TI - A morphological study of the brain of Solea senegalensis. I. The telencephalon. AB - In this paper we present an anatomical description of the telencephalon of Solea senegalensis based on cresyl violet and haematoxilin-eosin-stained serial transverse sections. This work was conducted as a basis for the precise localization of neuroendocrine territories in the brain of a species with growing interest in marine aquaculture. The external asymmetric morphology of Senegalese sole is correlated with the asymmetry of the forebrain. The right olfactory nerve and bulb are larger than the contralateral ones and this asymmetry is also extended to the cerebral hemispheres. The olfactory bulb comprises an outer olfactory nerve fiber layer, a glomerular layer, an external cellular layer, a secondary olfactory fiber layer and an internal cellular layer. The telencephalic hemispheres can be divided in area dorsalis and area ventralis, consisting of eleven and eight cell masses, respectively. The area dorsalis comprises five subareas: a pars medialis (Dm), subdivided into four nuclei termed Dml to Dm4; a pars dorsalis (Dd); a pars lateralis (D1), which consists of dorsal (Dld), ventral (Dlv) and posterior (Dlp) subdivisions; a pars centralis (Dc); and more caudally, a pars posterioris (Dp), which is very prominent in this species. A nucleus taenia (NT) was observed in the transitional region between area dorsalis and area ventralis. The area ventralis consists of pars dorsalis (Vd), pars ventralis (Vv), pars supracommissuralis (Vs), pars postcommissuralis (Vp), pars lateralis (V1), pars centralis (Vc), pars intermedia (Vi) and nucleus entopeduncularis (E). A periventricular organ, that we have termed lateral septal organ (LSO), was observed in the ventral telencephalon, medial to Vv. PMID- 10809355 TI - Morphological changes to somatotroph cells and in vitro individual GH release, in male rats treated with recombinant human GH. AB - The effect of in vivo chronic administration of recombinant human growth hormone (rhGH) on morphology and individual GH release in somatotroph cells was evaluated in young male Wistar rats. Over an 18-day period, 30-day-old male rats were injected daily with 1.5 1U rhGH/kg (GPG group) or saline (VPG group) by subcutaneous injection. Electron-immunocytochemical, ultrastructural and morphometric studies of somatotroph cells were carried out. Additionally, rat pituitary cells were dispersed and overall and individual GH release was studied by radioimmunoassay and cell immunoblot assay (quantified by image analysis), respectively. The ultrastructure and size of somatotroph cells did not change, but volume density of secretion granules was reduced (p<0.01) by previous in vivo GH treatment. At four days, basal GH release of rat pituitary cell monolayer cultures was lower in the GPG group than in the VPG group (p<0.05); after 12 hours of culture, GHRH stimulation of GH release was lower in the GPG group than in the VPG group (p<0.05), and GHRH+SRIH inhibited GH release in the GPG group (p<0.05), but not in the VPG group. The percentage of somatotroph cells was not modified, but the ratio of strongly/weakly GH-immunostained cells had changed; weakly GH-immunostained cells increased from 34% to 55%. Moreover, in vitro treatment with GHRH, SRIH, and both, easily changed the strongly/weakly GH immunostained cell ratio. Individual GH release, however, was not changed by previous in vivo GH treatment, although GHRH preferably stimulated a subpopulation of GH cells and SRIH did not inhibit individual GH release. These data suggest that exogenous chronic rhGH treatment down-regulates somatotroph function by modifying the proportion of GH cell subpopulation. PMID- 10809354 TI - Expression of c-kit and kit-ligand in benign and malignant prostatic tissues. AB - The tyrosine kinase receptor c-kit and its ligand [kit ligand (KL) or stem cell factor (SCF)] exert a broad range of biological activities during organogenesis and normal cell development. Recent studies have revealed that altered c-kit levels occur in a variety of malignancies and cancer cell lines. KL has also been shown to stimulate the growth of malignant cells, as well as to promote chemotaxis. We had previously reported expression of KL in stroma cells of normal human prostate. The present study was undertaken in order to analyze the patterns of expression of c-kit and KL in a well characterized set of prostatic tissues, including normal prostate (n=4), benign prostatic hyperplasia (BPH) (n=53) and adenocarcinoma (n=46) samples. The distribution of c-kit and KL proteins was studied by immunohistochemical analyses, while transcript levels were determined by in situ hybridization with specific RNA probes on a subset of the benign and malignant tissues referred above. In addition, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed to determine levels of c-kit and KL expression in cultures of epithelial and stroma cells, as well as in the prostate cancer cell lines LNCaP, DU145 and PC3. c-kit protein in normal prostate was exclusively detected in mast cells by immunohistochemistry and in situ hybridization. However, c-kit transcripts, but not c-kit protein, were detected in low levels and with an heterogeneous pattern in basal epithelial cells of ducts and acini. c-kit in BPH was detected in epithelial cells in 9 of 53 (17%) specimens. c-kit protein expression in malignant epithelial cells was identified in 1 of 46 (2%) tumors. However, c-kit transcripts were detected in low levels by in situ hybridization in most of the tumors analyzed. KL protein and transcripts in normal prostate were detected in high levels in stroma cells. However, epithelial cells were unreactive for anti-KL antibody, but showed low levels of KL transcripts mainly in cells of the basal layer. Basal epithelial cells in hyperplastic glands showed KL expression in 13 of 53 (24%) specimens. KL protein in tumor cells was noted in 18 of 46 (39%) cases. c-kit transcripts were not found in normal prostate and in the 3 cancer cell lines analyzed by RT-PCR, however, it was present in cultured epithelial cells of BPH, and in cultures of stroma cells from both normal and BPH. The majority of cultured cell lines of epithelial and stromal origin displayed considerable levels of KL. In addition all prostate cell lines studied showed significant levels of KL transcripts. In summary, co-expression of c-kit and KL in a subset of BPH cases may suggest an autocrine mode of signaling. Data from this study reveals that altered patterns of c-kit and KL expression are associated with BPH and adenocarcinoma of prostate. It appears that KL induces mast cells proliferation and maturation and enhances their release of protease. This could explain the accumulation of mast cells at tumor sites, a phenomenon that was not observed in normal prostate or BPH samples. PMID- 10809356 TI - Histochemical study of apoptotic epithelial cells depending on testosterone in primary cultured rat prostatic tissues. AB - To clarify whether apoptosis can be induced in cultured rat prostatic epithelial cells, they were investigated at various time points, depending on different concentrations of testosterone. Ventral lobes of rat prostates were cultured as small pieces of tissues up to 14 days. They were examined by anti-Fas antibody immunostaining and also compared to findings revealed by in situ end-labelling (ISEL) technique. To clarify apoptotic nuclei at high resolution, the quick freezing and deep-etching (QF-DE) method was also used, as reported before. The localization and appearance of Fas-positive cells were detected more widely and earlier than those of ISEL-positive cells, but both label-positive localizations were closely related to each other. In addition, they were detected more often in epithelial cells cultured with low testosterone concentrations. By the QF-DE method, chromatin fibers were found to be broken in spotty parts of apoptotic nuclei. We could control the concentration of testosterone in culture medium and detect the appearance of Fas antigen in cultured prostatic epithelial cells, followed by apoptotic changes. So, Fas and Fas-ligand system is one candidate for apoptosis in the prostate glands, depending on removal of hormonal testosterone. PMID- 10809357 TI - Characterization of inflammatory reaction in upper airways of cystic fibrosis patients. AB - Inflammatory cell populations have not been yet precisely evaluated in cystic fibrosis (CF) airways. We intended to characterize morphological modifications, inflammatory cell infiltration and cell proliferation in nasal tissues obtained from 15 CF patients and from 6 non-CF patients with nasal polyposis. Morphological analysis showed an intense inflammatory infiltration in CF and non CF tissues with only few modifications in the epithelium from CF tissues. Inflammatory cell populations characterized by specific immunolabeling were quantified, showing a predominance of macrophages and T- and B-lymphocytes and only moderate numbers of neutrophils in CF tissues; in non-CF polyps, lymphocytes and eosinophils were abundant. Proliferating cell percentages quantified after proliferating cell nuclear antigen immunolabeling were 5.3+/-4.1% (mean +/- SD) in CF polyps and 3.1+/-1.2% in non-CF polyps in epithelium but were very low in lamina propria. Intense inflammation in nasal tissues from CF patients is therefore dominated by macrophages and lymphocytes rather than by neutrophils. While morphology is preserved, proliferation is high in epithelium from CF polyps. These findings should be regarded in the future for a better understanding of inflammation in CF airway disease. PMID- 10809358 TI - The morphology of mesangial cells cultured at high density and in collagen gels. AB - Primary mesangial cells (rat) from monolayer cultures of the 6th to 12th passage and permanent SV40 Mes13 cells were grown at high density in organoid culture at the medium/air interphase. After adaptation to the in vitro conditions, both mesangial cell types developed after 7 days a synthesis apparatus (endoplasmic reticulum, Golgi apparatus) and produced matrix which consisted of Lamina densa like material, collagenous fibrils and filaments. Unspecific contacts, gap junctions and adhesion belts could be demonstrated in the contact areas. Additionally, some cells exhibited thick bundles of actin filaments. A close resemblance of the mesangial cells in high density culture to those in vivo can, therefore, be stated. Hence, they differentiated with regard to their matrix formation, contraction and contact behaviour and can therefore be used for experimental studies within a short culture period of 7 days. Cell aggregates in monolayer culture and in cultures in collagen gels had not differentiated at this stage. PMID- 10809359 TI - Neonatal treatment with monosodium glutamate (MSG): structure of the TSH immunoreactive pituitary cells. AB - Glutamic acid represents the most abundant stimulatory neurotransmitter in the central nervous system. Monosodium glutamate (MSG), subcutaneously administered to newborn rats in the perinatal period, induces lesions in 80 to 90% of the neurocytes of arcuate nuclei in the hypothalamus. These nuclei are the site of production of numerous stimulatory and inhibitory hormones including growth hormone releasing hormone (GHRH). The present studies were performed on male Wistar strain rats, subcutaneously injected on days 2, 4, 6, 8, and 10 of postnatal life with MSG at a dose of 4 mg/g body weight. Eighteen-month-old rats were additionally treated with Ambinon. When the animals reached the ages of 6 or 12 months, their body weight, body length and weight of pituitary were determined. On paraffin sections, using immunohistochemical techniques, TSH immunoreactive cells were detected and characterised by computerised image analysis. The results were subjected to statistical analysis using Student's t test. The rats which were perinatally treated with MSG and examined after 6 or 12 months of life were obese and shorter than control rats by 7% and 10% respectively. They also exhibited a reduction in the weight of the pituitary of 30% and 40% respectively in the two age groups. The proportion of TSH immunoreactive cells in the pituitary remained unchanged and amounted to 4.5% in the 6-month-old and 5.4% in the 12-month-old rats, respectively. The number of TSH-positive cells per mm2 area remained unchanged. The area and circumference of the cells in the 12-month-old rats were reduced by 22% and 18%, respectively. Perinatal injury to hypophyseal arcuate nuclei induced by monosodium glutamate injection, was not associated with any significant alterations in pituitary structure, as defined by the proportion of pituitary volume occupied by TSH immunoreactive cells. PMID- 10809360 TI - Transforming growth factor-beta1 gene and protein expression associated with atherogenesis of cholesterol-fed rabbits. AB - Transforming growth factor-beta1 (TGF-1beta) has been shown to modulate both cell proliferation and the synthesis of extracellular matrix by vascular cells. This study was aimed to establish the temporal correlation between TGF-beta1 expression, the expression of the extracellular matrix protein fibronectin, and plaque development during atherogenesis of hypercholesterolemic rabbits. New Zealand White rabbits were fed with 2% cholesterol-supplemented chow for 1 week, 2 weeks, 3 weeks or 6 weeks. TGF-beta1 mRNA and protein expression was examined in serial sections of aorta by in situ hybridization and immunohistochemistry. Fibronectin expression was examined by immunohistochemistry. In the control and 1 week feeding group, the expression of TGF-beta1 mRNA and protein was not apparent. In 2-week feeding group, intimal thickening was detected in which TGF beta1 mRNA and protein were not clearly observed, either. The 3-week and 6-week feeding groups exhibited fatty streaks in which TGF-beta1 mRNA and protein expression markedly increased as feeding proceeded. Cell type-specific staining indicated that TGF-beta1 was expressed by macrophages as well as smooth muscle cells of the fatty streaks. Immunostaining of fibronectin detected low expression levels in control, 1-week and 2-week feeding groups with pronounced upregulation in the thickened intima and the proximal media in 3-week and 6-week feeding groups. These results implicate a role for TGF-beta1 in modulating fatty streak formation and the synthesis of extracellular protein fibronectin during plaque development. PMID- 10809361 TI - Ultrastructural features of the gut in the white sturgeon, Acipenser transmontanus. AB - Electron-microscopic examinations of the sturgeon gut were performed. Oesophageal goblet cells were abundant in the stratified epithelium. The ultrastructural features of the secretory granules of the oesophageal and intestinal goblet cells were quite similar to those of other vertebrates. Lobules of multilocular adipose tissue were observed in the deep tunica propriasubmucosa of the oesophagus, in close association with vasculature and large fibre bundles of myelinated and unmyelinated axons. Similarly composed nerve fibre bundles were observed in the cardiac stomach, too. The presence of myelinated axons is an unusual feature in the vertebrate enteric nervous system. Cardiac and fundic zones of the stomach showed an epithelium with columnar ciliated and non-ciliated cells, the latter equipped with fuzzy microvilli. Cells lining the tubular gastric proper glands were markedly granulated. Intestinal superficial epithelium was columnar and contained ciliated, as well as non-ciliated and goblet cells. In the tunica propria all over the intestine, the presence and ultrastructure of granulated cells was in addition described. Intraepithelial granulated leukocytes were seen throughout the alimentary canal. Various types of endocrine cells were seen both in the stomach and in the intestine, the size of their granules was measured and their ultrastructure described and compared to that of mammalian cell types. PMID- 10809362 TI - Prolonged kallikrein inhibition does not affect the basal growth and secretory capacity of rat adrenal cortex, but enhances mineralo- and glucocorticoid response to ACTH and handling stress. AB - The effects on the pituitary-adrenocortical functions of the prolonged (7-day) blockade of endogenous bradykinin (BK) synthesis, obtained by the administration of the kallikrein inhibitor (K-I) cyclohexylacetyl-Phe-Arg-Ser-Val-Gln amide, were investigated in the rat. K-I treatment did not cause significant changes in the (i) body and adrenal weights; (ii) basal plasma levels of ACTH, aldosterone and corticosterone; and (iii) average volume of adrenocortical cells and their basal secretory capacity. Conversely, K-I administration induced a significant magnification of the in vivo mineralo- and glucocorticoid responses to the intraperitoneal (i.p.) bolus injection of ACTH. Moreover, K-I-treated rats, but not control ones, displayed a moderate and short-term adrenal secretory response to the mild stress evoked by the placebo i.p. injection. Collectively, these findings rule out the possibility that endogenous BK plays a relevant role in the control of adrenocortical function under basal conditions. However, they suggest that endogenous BK may be involved in quenching exceedingly high adrenocortical responses to ACTH and stresses. PMID- 10809363 TI - Expression of p53, p21/waf1, bcl-2, bax, Rb and Ki67 proteins in Hodgkin's lymphomas. AB - The aim was to investigate the combined immunoexpression of p53, p21, bcl-2, bax, Rb and Ki67 proteins in Hodgkin's lymphomas (HL) and correlate expression patterns with the histotype and the Epstein-Barr Virus (EBV) status. Paraffin sections from 56 cases of HL (18 nodular sclerosis and 38 mixed cellularity) and from ten "reactive" lymph nodes were investigated. P53, p21, bcl-2, bax, Rb and Ki67 proteins were detected in Hodgkin and Reed-Sternberg (HRS) cells in 35/56, 56/56, 24/56, 23/56, 56/56 and 56/56 cases of HL, respectively. No correlation was found between the expression of each protein and the EBV status or the histotype of HL. Comparison between p53 and p21 staining revealed two patterns: a) p53+/p21+ (35 cases); and b) p53-/p21+ (21 cases). The pattern p53+/p21+ suggests wild type p53 protein able to induce the expression of p21 while the p53 /p21+ pattern suggests p53-independent p21 expression. These results are consistent with the interpretation that inactivating p53 gene mutations may be rare in HL. Comparison between bcl-2 and bax staining showed a statistically significant relationship (p<0.001) for coexpression (19 cases) or absence of expression of both proteins (28 cases) in HRS cells. In contrast, bax expression was observed in most lymphoid cells in all "reactive" lymph nodes. Since the proapoptotic bax protein may act as tumour suppressor it is possible that the absence of this protein in HRS cells in a substantial proportion of HL may confer growth advantage and play a role in their pathogenesis. This could suggest bax gene alterations in some HL since in other studies acute lymphoblastic leukaemia cell lines demonstrate bax gene mutations with loss of bax immunoexpression. Another possibility is that reduced bax expression may be due to post transcriptional regulation, as was described in lymphoma cell lines. Comparison between Rb and Ki67 staining disclosed two main deviations from the normal parallel relationship in reactive lymph nodes: a) 2 cases with low Rb and high Ki67 expression possibly reflecting loss of Rb expression due to chromosome loss or to other abnormalities in the structure or the expression of Rb gene; and b) 9 cases with high RB and low Ki67 possible reflecting an attempt of Rb protein in excess to induce cell cycle arrest. Taken together, our findings provide combined immunohistological evidence for deregulated expression of cell-cycle and apoptosis-related proteins, that may play a role in the pathogenesis of HL. PMID- 10809364 TI - Comparative study of tumor angiogenesis and immunohistochemistry for p53, c ErbB2, c-myc and EGFr as prognostic factors in gastric cancer. AB - Gastric cancer (GC) continues to be a highly aggressive malignancy with poor prognosis and low survival rates. The survival of patients with GC depends mainly on the stage of the disease, with early GC having a 5 year survival of 90-100% and advanced tumors a 5 year survival of 15-25%. The role of other prognostic factors in these tumors is still under investigation. 28 gastric dysplasia, 45 Early GC and 98 Advanced Gastric Cancers were evaluated for expression of the oncogenes p53, c-ErbB2, c-myc and the EGFr in paraffin-embedded material utilizing Avidin-Biotin immunohistochemistry techniques. In 34 cases of GC microvessel density (MVD) was determined in CD34 stained sections. Statistical correlations with stage, histologic type, differentiation degree, location, size, ploidy patterns and overall survival were done. The Mantel-Cox test was performed to evaluate which factors had an independent prognostic value. Both, tumor angiogenesis and p53 protein expression were statistically associated (95% confidence intervals) with overall survival in patients with GC. p53 protein expression was also correlated with cardial location, nodal involvement and tumor stage. c-ErbB2 may recognize a group of highly aggressive well differentiated adenocarcinomas with worse prognosis. c-myc was also significantly enhanced in well differentiated tumors. EGFr showed no significant associations. Mantel-Cox was performed to compare the prognostic value of tumor stage, p53 protein expression and tumor angiogenesis. Tumor angiogenesis was the most important prognostic indicator to predict overall survival in our series. p53 expression was not independent and did not provide additional prognostic information to tumor stage. Our study suggests that angiogenesis as demonstrated by microvessel counts in CD34 stained sections is a significantly important prognostic factor for predicting survival in gastric cancer. PMID- 10809365 TI - Foreign serum-induced bile duct lesion (BDL) in athymic BALB/c nude mice. AB - To investigate a role of cellular immunity in foreign serum-induced bile duct lesion (BDL) in mice, athymic BALB/c nude (nu/nu) mice were intraperitoneally injected with swine serum (SS) twice a week up to 8 weeks and were compared with euthymic BALB/c heterozygote (nu/+) and wild-type (+/+) mice treated with SS in the same way for 4 weeks. All immunized nu/+ and +/+ mice developed marked BDL, and their sera showed high anti-SS IgE and IgG1 antibody titers, whereas no immunized nu/nu mice developed lesions, and their sera showed no elevation of antibody titers. Next, nu/nu mice were reconstituted with splenocytes derived from nu/+ mice, and then were intraperitoneally injected with SS twice a week for 3 weeks. Most of the reconstituted nu/nu mice developed BDL, and their sera showed the elevation of anti-SS IgE and IgG antibody titers. These results suggest that cellular immunity may play a pivotal role in the pathogenesis of swine serum-induced BDL. PMID- 10809366 TI - Effects of ethanol on the ultrastructure of the hamster thyroid C-cell. AB - The morphology of the thyroid C-cells in golden hamsters after short- and long term treatment with ethanol was studied. Immunohistochemistry was applied to examine the distribution of the C-cells in the thyroid gland. In the short-term experimental animals, the Golgi complexes and the granular endoplasmic reticulum were well developed and the number of the secretory granules was decreased as compared with those of the control animals. These findings suggest that the cellular activity of the thyroid C-cell is stimulated after short-term treatment with ethanol. The morphology of the thyroid C-cells of the long-term experimental animals was similar to that of the controls. It is conceivable that long-term treatment with ethanol does not affect the function of the C-cell. PMID- 10809367 TI - Cellular localization of fibroblast growth factor 2 (FGF-2) in benign prostatic hyperplasia. AB - Fibroblast growth factor 2 (FGF-2, basic fibroblast growth factor) has been reported to be elevated in tissues from benign prostatic hyperplasia (BPH), the most frequent neoplastic disease in aging men. This suggests that FGF-2 may play a significant role in the development of BPH. In this study the cellular distribution pattern of FGF-2 in tissues from BPH has been investigated by immunohistochemical and molecular biological methods. Radioimmunoassay revealed high concentrations of FGF-2, ranging between 450 and 950 ng per g tissue. Immunoblots confirmed the presence of a 18 kDa FGF-2 in tissue extracts. By immunohistochemistry done with a polyclonal antibody to recombinant FGF-2 on paraffin sections, FGF-2 was localized in fibroblasts, endothelial cells and smooth muscle cells of tissue samples of BPH. Nuclei of these cells were labelled distinctly. Moreover the cytoplasm of smooth muscle cells was labelled moderately. No immunostaining was seen in prostatic epithelium. Non-radioactive in situ hybridization with digoxygenin-labelled oligonucleotides revealed the presence of mRNA for FGF-2 in smooth muscle cells of the prostatic stroma. These results provide evidence that FGF-2 may be produced locally in the human prostate as a stroma-specific mitogen and may play a causal role in the development of BPH. PMID- 10809368 TI - A case study of ligation induced calcification in middle cerebral artery in rat. AB - A 90 min ligation of the middle cerebral artery (MCA) followed by 72-hour reperfusion appeared to cause calcium deposition in vascular myocytes of the tunica media and the perivascular tissue of the Sprague Dawley rat. The presence of small ovoid to large irregularly shaped intracellular opaque deposits were demonstrated by light and electron microscopy. Using X-ray elemental analysis the chemical nature of the deposits was found to be calcium phosphate. The functional significance of this first demonstration of acute calcification following transient ligation of the rodent MCA invites further studies. PMID- 10809369 TI - Brush-like cells within bronchial epithelia of chicken lung (Gallus gallus). AB - The secondary and primary (mesobronchus) bronchi of chicken lung are lined by a typical respiratory epithelium: pseudostratified columnar ciliated with goblet cells. Up to date, four constituting epithelial cell types have been identified: ciliated, mucosecretory, basal and endocrine cells. In this study a putative new epithelial cell type, the brush-like cell, is described. The avian brush-like cells have only been found in the bronchial epithelia but never in the gas exchange areas. They are scattered among the other epithelial cells, mainly ciliated cells, and their number is extremely low. The characteristic morphological feature of these cells is an apical protruding cytoplasm with microvilli. This cell type is similar to that found in the lung of some mammalian and non-mammalian species. The functional role of these cells is not yet clear; they could carry out absorptive processes. PMID- 10809370 TI - Intraepidermal free nerve fiber endings in the hairless skin of the rat as revealed by the zinc iodide-osmium tetroxide technique. AB - The nerve fiber distribution in the epidermis of the hairless rat skin was studied light microscopically by means of zinc iodide-osmium tetroxide staining. Two different morphological types of free nerve fiber endings could be detected: clusters of relatively thick nerve fibers stretched up through the spinous layer up to the granular layer sending off terminal branches. In addition, many solitary thin varicose nerve fibers were seen within the epidermis. The observed discrepancies in nerve fiber diameters appeared to be larger than those reported for human intraepidermal nerve fibers in recent immunohistochemical studies. Moreover, dendritic cells, most probably representing Langerhans cells, could be selectively stained. These cells appeared to be in a close location to thin varicose nerve fibers. Both types of demonstrated free nerve endings have to be functionally connected with different sensoric functions. Possibly, a subpopulation of the thin nerve fibers might possess primarily a nociceptive task, whereas the thick ones have most probably to be regarded as mechanoreceptive. The nerve fibers innervating dendritic cells appear to be identical to the peptidergic ones which may regulate the antigen-presenting capacity of these cells. Due to its selectivity for intraepidermal nerve fibers, the used method might supplement immunohistochemical procedures in a helpful manner. PMID- 10809371 TI - Mineralization of human premolar occlusal fissures. A quantitative histochemical microanalysis. AB - The mechanisms of cariogenesis in occlusal fissures remain elusive because of limited information about fissure structure and wall mineralization. The purpose of the present study was to determine the correlation between morphological patterns in occlusal fissures in human premolars and quantitative histochemical patterns of mineralization in the walls of these formations. We used scanning electron microscopy and quantitative X-ray microanalysis with the peak-to-local background ratio method and microcrystalline calcium salts as standards. We distinguished three morphological patterns of fissures in scanning electron microscopic images. The wall of the fissures was less mineralized than the control enamel in all three types of fissures. Because the fissure walls are hypomineralized, we suggest that practicing dentists should take into account the degree of mineralization when they are preparing the fissures for the application of sealant. PMID- 10809372 TI - Characterization of ligands for galectins, natural galactoside-binding immunoglobulin G subfractions and sarcolectin and also of the expression of calcyclin in thyroid lesions. AB - The purpose of this study was to characterize ligands for galectins, natural galactoside-binding immunoglobulin G subfractions and sarcolectin and also the expression of calcyclin in various benign and malignant thyroid lesions. The extent of the binding of eight glycochemical probes was quantitatively assessed using computer-assisted microscopy on 76 thyroid lesions including 10 not otherwise-specified multinodular goiters (S_MNG), 11 multinodular goiters with adenomatous hyperplasia (AH_MNG), 8 normomacrovesicular (NM_ADE) and 12 microvesicular (MIC_ADE) adenomas, and 9 papillary (P_CAR), 10 follicular variants of papillary (FvarP_CAR), 7 follicular (F_CAR) and 9 anaplastic (A_CAR) carcinomas. The 8 histochemical probes included 5 animal lectins (including galectins and sarcolectin), 1 polyclonal antibody (raised against calcyclin) and 2 immunoglobulin G subfractions from human serum with selectivity to alpha- and beta-galactosyl residues. The results show that multinodular goiters with adenomatous hyperplasia exhibited histochemical characteristics intermediate to those of normal multinodular goiters and microvesicular adenomas. Normomacrovesicular adenomas behaved very distinctly from microvesicular ones. Microvesicular adenomas were more closely related to differentiated thyroid carcinomas than any other type of benign thyroid lesions of epithelial origin. Papillary and follicular carcinomas seemed to represent the two extremes of the same biological entity with the follicular variant of the papillary carcinoma serving as a biological link between these two extremes. Anaplastic carcinomas behaved in a significantly different manner when compared to the differentiated forms of thyroid carcinomas. The results suggest that the patterns of expression of the glycoconjugates investigated in the present study may constitute useful tools for characterizing lesions in the human thyroid. PMID- 10809373 TI - Freeze-fracture cytochemistry: a new fracture-labeling method for topological analysis of biomembrane molecules. AB - Freeze-fracture cytochemistry allows visualization of cellular and molecular characteristics of biomembranes in situ. In this review, we discuss freeze fracture cytochemistry with special reference to a new cytochemical labeling of replicas, the detergent-digestion fracture-labeling technique. In this procedure, unfixed cells are rapidly-frozen, freeze-fractured, and physically stabilized by evaporated platinum/carbon. The frozen cells are then removed from the freeze fracture apparatus to thaw and are subsequently treated with detergents. After detergent-digestion, replicas are labeled with cytochemical markers. We demonstrate that the technique is a versatile tool for direct analysis of the macromolecular architecture of biomembranes and allows identification of particular intracellular membrane organelles. In addition, we demonstrate the application of ultrasmall gold to freeze-fracture immunocytochemistry. Freeze fracture cytochemistry is a valuable technique for investigating topology and dynamics of membrane molecules. PMID- 10809374 TI - Interferons and cell growth control. AB - Cytokines modulate cell growth, differentiation, and immune defenses in the vertebrates. Interferons (IFNs) are a unique class of cytokines that stimulate antiviral, antitumor and antigen presentation by inducing the expression of several cellular genes. Recent studies have identified a novel gene regulatory pathway activated by IFNs, which serves as a paradigm for most cytokine signal transduction pathways. A number of genes induced by IFNs participate in cell growth regulation and apoptosis. These include novel tumor suppressor genes. Although discovered as IFN-regulated factors, deletions of these genes cause leukemias in experimental models and in human patients. Genetic approaches have identified several novel regulators of apoptosis. Studies on the mechanism of action of these growth regulatory molecules are not only useful in identifying novel targets for the development of therapeutics but also help understand the molecular basis for loss of cell growth control and resistance to IFNs. This review focuses on the functions and roles of IFN regulated factors in cell growth control and mechanisms of disruption of IFN action in cancer cells. PMID- 10809375 TI - Linkage between cell membrane proteins and actin-based cytoskeleton: the cytoskeletal-driven cellular functions. AB - Asymmetric organization of the plasma membrane and cytosolic organelles is fundamental for a variety of cells, including bacteria, yeast and eukaryotic cells (Nelson, 1992). The degree into which cells polarize is characterized by their ability to create and maintain morphologically and biochemically distinct plasma membrane domains. The generation and maintenance of polarized distribution of membrane components (proteins and lipids) is thus critical to the ability of cells to perform complex activities such as cell-to-cell interactions, vectorial transport and secretion, cellular immunity, development and morphogenesis. Modification of cellular polarity may potentially lead to abnormal cellular activities and various pathological disorders (Molitoris, 1991; Carone et al., 1994; Chen et al., 1995). Our review shows the complex interplay between membrane proteins and the cytoskeletal network in determining the "polarized phenotype" in the cell. We provide evidence that membrane/cytoskeleton interaction is the key to regulation of the vast majority of cellular functions. PMID- 10809376 TI - Gain of function properties of mutant p53 proteins at the mitotic spindle cell cycle checkpoint. AB - Mutations in the p53 tumor suppressor gene locus predispose human cells to chromosomal instability. This is due in part to interference of mutant p53 proteins with the activity of the mitotic spindle and postmitotic cell cycle checkpoints. Recent data demonstrates that wild type p53 is required for postmitotic checkpoint activity, but plays no role at the mitotic spindle checkpoint. Likewise, structural dominant p53 mutants demonstrate gain-of function properties at the mitotic spindle checkpoint and dominant negative properties at the postmitotic checkpoint. At mitosis, mutant p53 proteins interfere with the control of the metaphase-to-anaphase progression by up regulating the expression of CKs1, a protein that mediates activatory phosphorylation of the anaphase promoting complex (APC) by Cdc2. Cells that carry mutant p53 proteins overexpress CKs1 and are unable to sustain APC inactivation and mitotic arrest. Thus, mutant p53 gain-of-function at mitosis constitutes a key component to the origin of chromosomal instability in mutant p53 cells. PMID- 10809377 TI - Vascular smooth muscle cell proliferation in the pathogenesis of atherosclerotic cardiovascular diseases. AB - Atherosclerosis is the principal cause of myocardial infarction, stroke, and peripheral vascular disease, accounting for nearly half of all mortality in developed countries. For example, it has been estimated that atherosclerosis leads to approximately 500,000 deaths from coronary artery disease and 150,000 deaths from stroke every year in the United States (American Heart Association, 1996). Percutaneous transluminal angioplasty has become a well-established technique for revascularization of occluded arteries. However, the long-term efficacy of the procedure remains limited by progressive vessel renarrowing (restenosis) within the following few months after angioplasty. Abnormal vascular smooth muscle cell (VSMC) proliferation is thought to play an important role in the pathogenesis of both atherosclerosis and restenosis. Accordingly, considerable effort has been devoted to elucidate the mechanisms that regulate cell cycle progression in VSMCs. In the present article, we will review the different factors that are involved in the control of VSMC proliferation, especially in the context of cardiovascular disease. Ultimately, a thorough understanding of these regulatory networks may lead to the development of novel drug and gene therapies for the treatment of cardiovascular diseases. Therapeutic approaches that targeted specific cell-cycle control genes or growth regulatory molecules which effectively inhibited neointimal lesion formation will be also discussed. PMID- 10809378 TI - Memory B cells and CD27. AB - Following antigen activation in germinal centers, B cells develop into memory B cells or plasma cells. Triggering via B-cell immunoglobulin receptors by antigens, cytokines and direct cell-to-cell contact by B and T cells plays an important role in the B cell differentiation into memory or plasma cells. Adult human peripheral blood B cells are separated into three subtypes by the expression of IgD and CD27, which belong to the tumor necrosis factor receptor (TNFR) family: IgD+ CD27- naive B cells, IgD+ CD27+ and IgD- CD27+ B cells. CD27+ B cells are larger cells with abundant cytoplasm carrying somatic hypermutation, and have an ability to produce immunoglobulin, indicating that CD27 is a memory marker of B cells. The ligation of CD27 yields crucial signals that positively control the entry of B cells into the pathway to plasma cells. We review observations on subpopulations and differentiation of mature B-cells by T/B cell interaction via CD27/CD70 as compared with CD40/CD154 interaction, and discuss about memory B cells. PMID- 10809379 TI - Tenascin in the developing and adult human intestine. AB - The tenascins are a family of multifunctional extracellular matrix glycoproteins subject to complex spatial and temporal patterns of expression in the course of various organogenetic processes, namely those involving epithelial-mesenchymal interactions. In the intestine, the tenascins, in particular tenascin-C, have been found to be differentially expressed in the developing and adult small intestinal and colonic mucosa as well as in neoplasm. While tenascin-C emerges as a key player likely to be involved in intestinal mucosa development, maintenance and disease, its exact role in the regulation of fundamental intestinal cell function(s) such as proliferation, migration and tissue-specific gene expression remains however to be established. PMID- 10809380 TI - Abnormal distribution of CD45 isoforms expressed by CD4+ and CD8+ T cells in rheumatoid arthritis. AB - CD45RO+ T cells are referred to as memory or helper-inducer while CD45RA+ T cells are regarded as naive or suppressor-inducer T cells. The former population predominates in the peripheral blood and even more in the synovial fluid of patients with rheumatoid arthritis, to the expense of the latter population. Within the CD45RB+ compartment, there appears to be more of the fully differentiated than of the early-differentiated CD4+ T cells. In spite of the fact that these lymphocytes are close to undergoing apoptosis, this programmed cell death is inhibited in the rheumatoid synovium. PMID- 10809381 TI - Schwann cell extracellular matrix molecules and their receptors. AB - The major cellular constituents of the mammalian peripheral nervous system are neurons (axons) and Schwann cells. During peripheral nerve development Schwann cells actively deposit extracellular matrix (ECM), comprised of basal lamina sheets that surround individual axon-Schwann cell units and collagen fibrils. These ECM structures are formed from a diverse set of macromolecules, consisting of glyco-proteins, collagens and proteoglycans. To interact with ECM, Schwann cells express a number of integrin and non-integrin cell surface receptors. The expression of many Schwann cell ECM proteins and their receptors is developmentally regulated and, in some cases, dependent on axonal contact. Schwann cell ECM acts as an organizer of peripheral nerve tissue and strongly influences Schwann cell adhesion, growth and differentiation and regulates axonal growth during development and regeneration. PMID- 10809382 TI - Id genes in nervous system development. AB - Id genes encode helix-loop-helix proteins that function to mediate processes important for normal development including cellular differentiation, proliferation and apoptosis. Id proteins act as negative regulators of other transcription factors, which are essential for cell determination and differentiation in diverse cell types, and interact with proteins important for cell cycle regulation. Studies of Id gene expression in the nervous system and in neural cells in culture indicate that Id proteins contribute to the regulation of mammalian nervous system development. Also, recognition of a wide variety of proteins with which Id transcription factors are capable of interacting suggests that it will be possible to understand more precisely their specific functions and importantly how these are integrated. PMID- 10809383 TI - Pathophysiology of primary hyperparathyroidism. AB - Parathyroid gland is the overall regulatory organ within the systemic calcium homeostasis. Through cell surface bound calcium-sensing receptors external calcium inversely regulates release of parathyroid hormone (PTH). This mechanism, which is voltage independent and most sensitive around physiologic calcium concentrations, is regulated through a 120 kDa calcium sensing receptor, CaR. Inherited inactivation of this receptor is the cause for familial hypocalciuric hypercalcemia (FHH). Parallel research identified the 550 kDa glycoprotein megalin, which also is expressed on the parathyroid cell surface, as another potential calcium sensing protein. Although this protein expresses numerous calcium binding sites on its external domain, its main function may be calcium sensitive binding and uptake of steroid hormones, such as 25-OH-vitamin D3 (bound to vitamin D binding protein) and retinol. In hyperparathyroidism (HPT), excessive PTH is secreted and the calcium sensitivity of the cells reduced, i.e. the set-point, defined as the external calcium concentration at which half maximal inhibition of PTH release occurs, shifted to the right. Pathological cells have reduced expression of both CaR and megalin, and reduced amount of intracellular lipids, possibly including stored steroid hormones. A number of possible genetic disturbances have been identified, indicating multifactorial reasons for the disease. In postmenopausal women, however, the individual group with highest incidence of disease, a causal relation to reduced effect of vitamin D is possible. An incipient renal insufficiency with age, lack of sunshine in the Northern Hemisphere, and an association to the baT haplotype of the vitamin D receptor supports this theory. This review summarizes data on regulation of PTH release, dysregulation in HPT, as well as proliferation of parathyroid cells. PMID- 10809384 TI - Are hyaluronan receptors involved in three-dimensional cell migration? AB - Hyaluronan (HA), an unbranched polysaccharide consisting of repeated glucuronic acid/N-acetylglucosamine disaccharide units, is ubiquitously present in the extracellular matrix of many tissues (for a more comprehensive review see: Fraser et al., 1997). Increased amounts of hyaluronan are produced by solid tumors and tumor-associated fibroblasts, and tumor-induced HA is correlated with poor prognosis. HA is well known to stimulate the migration of a large variety of cell types. Stimulation of cell migration by HA has been explained by different mechanisms. HA was shown to specifically bind to cell surface receptors, and inhibition of HA-receptor function was demonstrated to decrease cell migration and tumor growth. On the other hand, HA as a large hydrophilic molecule is also known to modulate the extracellular packing of collagen and fibrin, leading to increased fiber size and porosity of extracellular substrates. Hence a modified matrix architecture might similarly account for increased locomotion of cells. In this review, we attempted to summarize the available data on HA-induced cell migration, with particular emphasis on the role of HA receptors in three dimensional cell migration. Although the HA receptor CD44 has been shown to mediate migration of cells over two-dimensional hyaluronan-coated surfaces in vitro, there is only little evidence that HA-binding to CD44 or other HA receptors has major impact on the locomotion of cells through three-dimensional matrices in vivo. We showed recently that the promigratory effect of HA in fibrin gels is largely due to HA-mediated modulation of fibrin polymerization. By increasing the porosity of fibrin gels, HA strongly accelerates cell migration. The porosity of matrices therefore appears as an important and probably underestimated determinant of cell migration and tumor spread. PMID- 10809385 TI - Melatonin, experimental basis for a possible application in breast cancer prevention and treatment. AB - The role of the pineal as an oncostatic gland has been studied in animal models of tumorigenesis, especially on those concerning the mammary gland. The general conclusion is that experimental manipulations activating pineal gland, or the administration of melatonin, reduce the incidence and growth rate of chemically induced murine mammary tumors, while pinealectomy or situations which implicate a reduction of melatonin production usually stimulate mammary carcinogenesis. The direct actions of melatonin on mammary tumors have been suggested because of its ability to inhibit, at physiological doses (1nM), the in vitro proliferation of MCF-7 human breast cancer cells. In this article we review the outstanding findings related to melatonin actions on mammary which, taken together, support a possible usefulness of this indoleamine in the prevention and treatment of mammary gland malignancy. PMID- 10809386 TI - Skeletal muscle development in the mouse embryo. AB - In this review we discuss the recent findings concerning the mechanisms that restrict somitic cells to the skeletal muscle fate, the myogenic regulatory factors controlling skeletal muscle differentiation and specification of myogenic cell lineages, the nature of inductive signals and the role of secreted proteins in embryonic patterning of the myotome. More specifically, we review data which strongly support the hypothesis that Myf-5 plays a unique role in development of epaxial muscle, that MyoD plays a unique role in development of hypaxial muscles derived from migratory myogenic precursor cells, and that both genes are responsible for development of intercostal and abdominal muscles (hypaxial muscles that develop from the dermatomal epithelia). In addition, while discussing upstream and post-translational regulation of myogenic regulatory factors (MRFs), we suggest that correct formation of the myotome requires a complex cooperation of DNA binding proteins and cofactors, as well as inhibitory function of non-muscle cells of the forming somite, whose proteins would sequester and suppress the transcription of MRFs. Moreover, in the third part of our review, we discuss embryonic structures, secreted proteins and myogenic induction. However, although different signaling molecules with activity in the process of somite patterning have been identified, not many of them are found to be necessary during in vivo embryonic development. To understand their functions, generation of multiple mutants or conditional/tissue-specific mutants will be necessary. PMID- 10809387 TI - Role of diffusible and transcription factors in inner ear development: implications in regeneration. AB - Organogenesis involves a dynamic balance of the mechanisms regulating cell division, differentiation and death. The development of the chicken embryo inner ear offers a well-characterised model at the morphological level to study which signals are implicated in the modulation of cellular activation and commitment. The early developmental decisions that control the origin of the inner ear elements are just beginning to be identified by complementary in vivo and in vitro studies. Insulin-like growth factor-I (IGF-I) and nerve growth factor (NGF) are among the best characterised diffusible factors acting during inner ear development. Although the cellular actions of these factors are beginning to be understood, the signalling pathways triggered by them still remain largely unknown. In this context, viral vehicles can be used to deliver genes and then analyse their functional roles during inner ear development. A model is proposed where the actions of IGF-I and NGF contribute to the combinatorial expression of Jun and Fos family members in particular domains of the otic vesicle. Some of these mechanisms may be also implicated in otic regeneration. PMID- 10809388 TI - New sequences and new fungal producers of peptaibol antibiotics antiamoebins. AB - Mixtures of the microheterogeneous 16-mer peptaibol antibiotics called antiamoebins (AAM) have been isolated from the culture broths of strains of the filamentous fungi Stilbella erythrocephala ATCC 28144, Stilbella fimetaria CBS 548.84 and Gliocladium catenulatum CBS 511.66. Sequences were determined using on line HPLC together with positive- and negative-ion electrospray ionization mass spectrometry. Some characteristic features are recognized in the mass spectrometric fragmentation pattern of AAM. From a sample originally used for sequencing AAM (from Hindustan Antibiotics, Ltd., Pimpri, Poona-411018, India), and a sample of AAM commercially available (from Sigma Chemicals, St. Louis, MO, USA) HPLC elution profiles and sequences were assigned. Further, sequences of AAM previously isolated from Emericellopsis synnematicola CBS 176.60 and Emericellopsis salmosynnemata CBS 382.62 were determined. The peptide designated AAM I was the most abundant in all isolates and its structure could be confirmed. AAM II was detectable as a minor component (1.9%) only in the original sample of AAM, but not in the other isolates. The structures of AAM III, IV and V, which had previously been partly assigned, were definitely established, and the new sequences AAM VI-XVI were elucidated. AAM showing Phe1/Leu1 or Phe1/Val1 exchange, respectively, are produced in amounts only by S. erythrocephala. Sequences, HPLC elution profiles ('fingerprints') and relative amounts of peptides of all isolates were correlated. PMID- 10809389 TI - GABA-induced changes of the tissue-specific peptide pool of white rat brain. AB - Internasal administration of gamma-aminobutyric acid (GABA) induced prolonged behaviour changes and the appearance of three new compounds absent in the brain extracts of control rats. Two peptides associated with GABA administration were isolated and sequenced: Thr-Tyr-Thr-Phe, which corresponds to a gamma immunoglobulin segment, and Val-Leu, which is present in a great number of proteins, hence its precursor could not be established. The third compound was not amenable to the Edman degradation technique. The data obtained show that the introduction of a neurotransmitter could cause specific changes in the levels of tissue-specific peptide components. PMID- 10809390 TI - Synthesis and antibody recognition of mucin 1 (MUC1)-alpha-conotoxin chimera. AB - We synthesized and characterized new chimera peptides by inserting an epitope of the mucin 1 glycoprotein (MUC1) as a 'guest' sequence in the 'host' structure of alpha-conotoxin GI, a 13-residue peptide (ECCNPACGRHYSC) isolated from the venom of Conus geographus. The Pro-Asp-Thr-Arg (PDTR) sequence of MUC1 selected for these studies is highly hydrophilic and adopts a beta-turn conformation. The alpha-conotoxin GI also contains a beta-turn in the 8-12 region, which is stabilized by two disulphide bridges in positions 2-7 and 3-13. Thus, the tetramer sequence of alpha-conotoxin, Arg9-His-Tyr-Ser12, has been replaced by PDTR, comprising the minimal epitope for MUC1 specific monoclonal antibodies (MAbs) HMFG1 (PDTR) and HMFG2 (DTR). Synthesis of the chimera peptide was carried out by Fmoc strategy on (4-(2',4'-dimethoxyphenyl-aminomethyl)phenoxy) (Rink) resin and either 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB) or air oxidation was applied for the formation of the first Cys3-Cys13 or Cys2-Cys7 disulphide bridge, respectively. For the second disulphide bridge, three different oxidation procedures (iodine in acetic acid, 10% DMSO/1 M HCl or tallium trifluoroacetate (Tl(tfa)3) in TFA) were utilized. The HPLC purified peptides were characterized by electrospray mass spectrometry (ES-MS) and amino acid analysis. The CD spectra of the bicyclic MUC1-alpha-[Tyr1]-conotoxin chimera peptide showed partially ordered conformation with turn character. In antibody binding studies, the RIA data showed that both the linear and the bicyclic forms of MUC1-alpha-[Tyr1] conotoxin chimera were recognized by MAb HMFG1 specific for PDTR sequence, while no binding was observed between MAb HMFG2 and various forms of the chimera. MAb HMFG1, using synthetic epitope conjugates or native MUC1 as target antigens, recognizes the PDTR motif more efficiently in the linear than in the bicyclic compound, but no reactivity was found with the monocyclic forms of MUC1-alpha [Tyr1]-conotoxin chimera, underlining the importance of certain conformers stabilized by double cyclization. PMID- 10809391 TI - Predominant torsional forms adopted by dipeptide conformers in solution: parameters for molecular recognition. AB - The present paper describes the predominant conformational forms adopted by dipeptides in aqueous solution. More than 50 dipeptides were subjected to conformational analysis using SYBYL Random Search. The resultant collections of conformers for individual dipeptides, for small groups with related side chain residues and for large groups of about 50 dipeptides were visualized graphically and analysed using a novel three-dimensional pseudo-Ramachandran plot. The distribution of conformers, weighted according to the percentage of each in the total conformer pool, was found to be restricted to nine main combinations of backbone psi (psi) and phi (phi) torsion angles. The preferred psi values were in sectors A7 (+150 degrees to +/-180 degrees), A10 (+60 degrees to +90 degrees) and A4 (-60 degrees to -90 degrees), and these were combined with preferred phi values in sectors B12 (-150 degrees to +/-180 degrees), B9 (-60 degrees to -90 degrees) and B2 (+30 degrees to +60 degrees). These combinations of psi and phi values are distinct from those found in common secondary structures of proteins. These results show that although dipeptides can each adopt many conformations in solution, each possesses a profile of common conformers that is quantifiable. A similarly weighted distribution of dipeptide conformers according to distance between amino-terminal nitrogen and carboxyl-terminal carbon shows how the preferred combinations of backbone torsional angles result in particular N-C geometries for the conformers. This approach gives insight into the important conformational parameters of dipeptides that provide the basis for their molecular recognition as substrates by widely distributed peptide transporters. It offers a basis for the rational design of peptide-based bioactive compounds able to exploit these transporters for targeting and delivery. PMID- 10809392 TI - Weaning food and magadi associated with dental fluorosis in Northern Tanzania. AB - A cross-sectional study of 143 children aged 10-14 years was carried out to identify factors associated with the severity of dental fluorosis in 2 areas (Moshi/Kibosho: n = 63/80) with fluoride (F) concentrations < 0.4 mg F/L in the drinking water. Dental fluorosis was recorded under field conditions using the Thylstrup and Fejerskov index (TFI). The score on tooth 21 was used to indicate the severity. The prevalence of dental fluorosis in Moshi at TFI score > or =1 was 60% and at TFI score > or =5 it was 10%. The corresponding values in Kibosho were 100% and 34%, respectively. Background variables pertained primarily to the child's first 6 years of life. In Moshi and Kibosho, 65% and 97% of children, respectively, used magadi, a fluoride-containing food tenderizer. In Moshi, the risk of having TFI score > or =2 was significantly higher among users of magadi (odds ratio (OR) = 5). Kiborou, a traditional homemade weaning food (porridge) cooked with magadi, was used only in Kibosho. Users of kiborou (36%) were at significantly higher risk (OR = 3) of developing fluorosis at severity TFI > or =4 than the users of lishe, another type of weaning food. All children in Moshi and 64% in Kibosho used lishe, which is cooked without magadi. Magadi and kiborou were significant risk indicators. Thus, it seems that the unexpectedly high prevalence of dental fluorosis and the observed differences in fluorosis prevalence and severity may be partly explained by F exposure from magadi. PMID- 10809393 TI - Salivary buffer effect in relation to late pregnancy and postpartum. AB - We studied the salivary pH, buffer effect (BE), and flow rates of unstimulated and paraffin-stimulated saliva of 8 women in their late pregnancy and postpartum. Salivary samples were collected about 1 month prior to and about 2 months after delivery. In non-pregnant control women, two paraffin-stimulated salivary samples were collected 1 month apart. The salivary BE increased significantly from late pregnancy to postpartum without exception. The increase was 2.04 +/- 1.17 pH units (P < 0.001) on average. The BE increased from 4.79 +/- 1.64 (final pH) to 6.82 +/- 1.01 (final pH). This change was not due to variation in salivary flow rates, since both unstimulated and paraffin-stimulated flow rates remained unchanged. In control women the difference between the 2 BE measurements was only 0.13 +/- 0.47 pH units on average. We concluded that women with high postpartum BE values may have moderate or even low BE values in late pregnancy. In control women, individual variation was found to be low in all variables studied. PMID- 10809394 TI - Knowledge, beliefs and behavior related to oral health among Tanzanian and Ugandan teacher trainees. AB - This study was designed to assess levels of oral health knowledge, beliefs and self-reported behavior among teacher trainees in Uganda and Tanzania. Anonymous questionnaire data were collected from trainees attending the final year at teacher training colleges in Rungwe district, Tanzania, in 1997, and in Mbale and Kampala districts of Uganda, in 1998. A total of 195 trainees from Tanzania and 225 trainees from Uganda participated. Results from Pearson's chi2 test and independent sample t tests indicated that Tanzanian students had less experience with oral impairments, were more resolutely prepared to teach about the importance of personal habits for oral health maintenance, and had higher levels of oral health knowledge than Ugandan students. Ugandan students endorsed frequent consumption of sugar products more often than their Tanzanian counterparts, but they were more diligent in visiting dentists. Most of the Tanzanian (97.4%) and Ugandan (95.6%) students reported daily toothbrushing. In both countries, beliefs about the importance of preventive behaviors for oral health were closely related to the frequency with which such habits occurred. This cross-cultural consistency highlights the importance of cognitive factors. The implications for oral health educational programs among teacher trainees across East African countries are discussed. PMID- 10809395 TI - A Swedish version of the Dental Visit Satisfaction Scale. AB - The aims of this study were to translate and analyze the Dental Visit Satisfaction Scale (DVSS), which has been developed to measure different aspects of the dentist-patient relationship in the view of the patient. Subjects were ordinary dental patients attending dental care in various public dental service clinics (PDS) and patients attending an emergency PDS clinic and an oral medicine clinic (n = 204). Consecutively, patients were asked to answer a questionnaire that consisted of the DVSS and information about dental anxiety, age, and gender. The results revealed similar DVSS item/sum of scores levels as those in previous studies. There was no significant difference with respect to gender. High dental anxiety was associated with low DVSS score. Three dimensions were found in the exploratory factor analysis: Information/communication, Understanding/acceptance, and Technical competence. Item 8 was found to have skewed properties according to correlation, reliability, and factor analysis. A confirmatory factor analysis with the 9-item DVSS (item 8 removed) revealed a model with 4 dimensions. A general dental satisfaction factor was found, loading on all 9 items, together with the above-mentioned, more narrow factors. PMID- 10809396 TI - Periodontal status of adult Sudanese habitual users of miswak chewing sticks or toothbrushes. AB - Miswak chewing sticks are prepared from the roots or twigs of Salvadora persica plants. They are widely used as a traditional oral hygiene tool in several African and Middle Eastern countries. The aim of this study was to assess and compare the periodontal status of adult Sudanese habitual miswak and toothbrush users. The study population comprised male miswak users (n = 109) and toothbrush users (n = 104) with age range 20-65 years (mean 36.6 years) having 18 or more teeth present. They were recruited among employees and students at the Medical Sciences Campus in Khartoum, Sudan. One examiner used the Community Periodontal Index (CPI) to score gingival bleeding, supragingival dental calculus, and probing pocket depth of the index teeth of each sextant. In addition, the attachment level was measured, which, along with the CPI, was used to assess the periodontal status of the two test groups. Gingival bleeding and dental calculus were highly prevalent in the study population. Approximately 10% of the subjects had > or =4 mm probing depth and 51% had > or =4 mm attachment loss in one or more sextants. Subjects in the age group 40-65 years had a significantly (p < 0.05) higher number of sextants with gingival bleeding and with > or =4 mm probing depth and attachment loss than the 30-39 years group. Miswak users had significantly (p < 0.05) lower dental calculus and > or =4 mm probing depth and higher > or =4 mm attachment loss as well as a tendency (p = 0.09) to lower gingival bleeding in the posterior sextants than did toothbrush users. These differences were not significant in the anterior sextants. It is concluded that the periodontal status of miswak users in this Sudanese population is better than that of toothbrush users, suggesting that the efficacy of miswak use for oral hygiene in this group is comparable or slightly better than a toothbrush. Given the availability and low cost of miswak, it should be recommended for use in motivated persons in developing countries. PMID- 10809397 TI - Effect of acid-etching on remineralization of enamel white spot lesions. AB - This in vitro study aimed at investigating whether full remineralization would occur in white spot lesions when the surface porosity was increased by acid etching. The effect of fluoride was also investigated. Enamel blocks with in vitro produced white spot lesions were used. Group A was exposed to a remineralizing solution only. In group B, the lesions were etched with 35% phosphoric acid for 30 s, then treated as in group A. Group C was treated as group A + daily treatment with a fluoride toothpaste slurry (1,000 ppm) for 5 min. Group D was treated as group B + the daily fluoride treatment of group C. The remineralization was measured weekly with Quantitative Light-induced Fluorescence during the experimental period. After 10 weeks of remineralization, mineral profiles were assessed with transverse microradiography. The enamel fluorescence was partly regained. There were significant differences in the lesion depth, mineral content at the surface layer, and integrated mineral loss between the groups. Addition of fluoride accelerated the remineralization only in the beginning; in later stages the process leveled out and even reached a plateau in all the groups. It was concluded that full remineralization was not achieved by etching, by the addition of fluoride, nor by the combination of both treatments in this in vitro study. PMID- 10809398 TI - Psychosocial aspects of dental and general fears in dental phobic patients. AB - Phobic reactions can often be a significant health problem for fearful dental patients. This is true in particular for individuals with long-time avoidance and elevated general psychological distress. The aim of this study was to investigate the expression of dental fear among 67 dental phobic individuals with a low versus high degree of general fear. Reported etiologic background factors, avoidance time, general psychological distress and psychosocial manifestations and consequences of dental phobia were studied. No significant difference in dental anxiety level between individuals with low versus high general fear was found, and both groups reported high frequencies of negative dental experiences. The low-fear group reported a longer (though not statistically significant) average avoidance time than the high-fear group. However, patients with a high level of general fear showed a significantly higher degree of psychological distress, and also reported stronger negative social consequences from their dental anxiety. These results indicate that the character of dental fear might be different between different groups of fearful patients, which makes the condition psychologically handicapping. Such aspects should be assessed in the diagnostic analysis of patients with severe dental anxiety. In particular, assessments of signs of general psychological distress are warranted. PMID- 10809399 TI - In vitro shear bond strength of orthodontic bondings without liquid resin. AB - This study aimed at evaluating the early shear bond strength of enamel-composite bracket adhesion accomplished without the use of liquid resin. Orthodontic brackets were bonded to the buccal surfaces of healthy extracted premolars in the test group by Transbond XT (n = 8) and Phase II (n = 8) composites but not the enclosed liquid resins in these products. Brackets bonded with the same materials (n = 8 for each) along with their corresponding liquid resin served as controls. The specimens were tested for shear bond strength after 24-h storage in water at 37 degrees C. The fractured surfaces were graded with Adhesive Remnant Index (ARI) under a 2x-dissection microscope. Enamel of the randomly selected test and control specimens was dissolved by 20% formic acid. Afterwards, the enamel side of the bonding materials in both groups (n = 4) was examined under the scanning electron microscope. ANOVA was used for statistical analyses. Our laboratory data suggest that the enamel adhesion produced by these two commercial materials without the use of liquid resin does not differ significantly in their early in vitro shear bond strength. PMID- 10809400 TI - Cell death mechanisms in Parkinson's disease. AB - OBJECTIVE: While the causes of neuronal death in Parkinson's disease (PD) and other neurodegenerative disorders are still unknown, several mechanisms are under discussion: programmed vs. passive cell death (apoptosis vs. necrosis), mainly based on conflicting results on the rare presence or absence of DNA fragmentation in substantia nigra neurons using the in situ DNA-labeling (TUNEL) method. DESIGN/METHODS: In 4 cases of Parkinson's disease (PD), 2 cases of Dementia with Lewy bodies (DLB) and 3 age-matched controls, the TUNEL/ISEL method was used to detect DNA fragmentation in substantia nigra locus coeruleus and cerebral cortex [method by Gold et al. (1994)]. In addition, immunohistochemistry was performed for an array of apoptosis-related proteins, i.e. the recently described apoptosis specific protein cJun/AP1 (ASP), the proto-oncogenes c-Jun, c-Jun AP1, Bcl2, Bax, Bcl-x, p53, CD 95 (Fas/Apo-1), activated caspase 3, several heat shock proteins (alpha-B crystallin, ubiquitin), and alpha-synuclein. RESULTS: None of the cases of PD, DLB, and controls showed convincing TUNEL-positivity nor morphologic signs of apoptosis in nigral, locus coeruleus or cortical neurons with or without Lewy bodies but variable numbers of TUNEL-positive astrocytes and microglial cells in substantia nigra of PD and DLB. There were no significant differences in the expression of c-Jun, ASP, Bcl-2, Bax, and Bcl-x in substantia nigra neurons between PD, DLB, and controls nor between cortical and subcortical neurons with and without Lewy bodies. No expression of p53, and activated caspase 3, or any of the examined stress proteins was seen in neurons, while reactive astroglia and microglia were decorated by antibodies to Bcl-2, Bax, alpha-B-crystallin and less, to Bcl-x and caspase 3. Lewy bodies, dystrophic neurites and axonal spheroids, all being negative for the applied apoptosis regulating proteins, showed strong expression of the examined stress proteins and of alpha-synuclein. CONCLUSIONS: These findings which are in line with previous results in Alzheimer's disease (Stadelmann et al., 1998) and Parkinson's disease (Banati et al., 1999) suggest that mechanisms distinct from classical apoptosis play a central role in the pathogenesis of PD and related neurodegenerative diseases. Further studies are warranted to elucidate the intracellular cascade of events leading to cell death in these disorders showing slow progression over many years. PMID- 10809401 TI - Involvement of alpha-synuclein in Parkinson's disease and other neurodegenerative disorders. AB - A major step in the elucidation of the pathogenesis of neurodegenerative disorders was the identification of a mutation in the alpha-synuclein gene in autosomal dominant Parkinson's disease (PD). Alpha-synuclein is the main component of Lewy bodies (LB), the neuropathological hallmark of PD. Moreover, a fragment of alpha-synuclein (NAC) is the second major component of amyloid plaques in Alzheimer's disease (AD). Recent studies of other neurodegenerative disorders such as dementia with LB (DLB), multiple system atrophy (MSA) and amyotrophic lateral sclerosis (ALS) also revealed intracellular accumulations of alpha-synuclein in affected brain regions. This may indicate that these disorders partially share common pathogenic mechanisms. Recent data provide first insights into the physiological function of alpha-synuclein and support the concept of an essential role of alpha-synuclein in neurodegeneration. Increasing knowledge on the pathogenic molecular mechanisms of neurodegeneration and of the pathophysiological function of alpha-synuclein in particular may influence future development of therapeutic strategies in neurodegenerative disorders. PMID- 10809402 TI - Altered redox state of platelet coenzyme Q10 in Parkinson's disease. AB - BACKGROUND: The reduced form of coenzyme Q10 (CoQ10) acts as a lipophilic antioxidant and participates in electron and proton transport of the respiratory chain in the inner mitochondrial membrane. An alteration in CoQ10 redox state may thus reflect a change in membrane electron transport and the effectiveness of defense against toxic reactive oxygen species such as hydrogen peroxide and superoxide. In Parkinson's disease alterations in the activities of complex I have been reported in substantia nigra and platelets. Deficiency of mitochondrial enzyme activities could affect electron transport which might be reflected by the platelet CoQ10 redox state. METHOD: We have determined concentrations of the reduced and oxidized forms of CoQ10 and the activity of monoamine oxidase B in platelets isolated from parkinsonian patients and age- and gender-matched controls. RESULTS: Platelet CoQ10 redox ratios (reduced CoQ10 to oxidized CoQ10) and the ratio of the reduced form, compared with total platelet CoQ10, were significantly decreased in de novo parkinsonian patients. Platelet CoQ10 redox ratios were further decreased by L-DOPA treatment (not significant), whilst selegiline treatment partially restored CoQ10 redox ratios. Monoamine oxidase activities in non-selegiline treated patients were similar to controls. INTERPRETATION: Our results either suggest an impairment of electron transport or a higher need for reduced forms of CoQ10 in the platelets of even de novo parkinsonian patients. However, the CoQ10 redox ratio was not correlated to disease severity, as determined by the Hoehn and Yahr PD disability classification, suggesting that this parameter may not be useful as a peripheral trait marker for the severity of PD but as an early state marker of PD. PMID- 10809403 TI - Evidence for impaired presynaptic dopamine function in parkinsonian patients with motor fluctuations. AB - We used [18F]6-fluorodopa (FD) positron emission tomography (PET) to examine the severity of nigrostriatal dopaminergic dysfunction in 67 patients with Idiopathic Parkinsonism (IP), 52 with fluctuations and 15 with a stable response to levodopa. FD uptake (Ki) was reduced by 12% in the caudate (p = 0.08) and by 28% in the putamen (p = 0.0004) of patients with fluctuations compared to those with a stable response. However, there was considerable overlap of FD Ki values between the two groups. The fluctuators had a longer symptom duration (11.6 +/- 5.7 years) than the patients with a stable response to levodopa (4.3 +/- 2.4 years; p < 0.0001) and the age of onset of symptoms was earlier in the fluctuators (43.9 +/- 8.9 versus 54.1 +/- 10.4; p = 0.0004). Similar reductions in FD Ki in the fluctuators persisted following adjustment for these variables (7.5% in the caudate and 26% in the putamen; p = n.s. and 0.007, respectively). When smaller groups (n = 15 each) were matched for duration of symptoms, the reduction in caudate Ki in the fluctuators was only 1.9% (p = n.s.), but there was still a 24% reduction in putamen Ki (p = 0.05). These findings suggest that fluctuators and non-fluctuators may differ in the severity of their nigrostriatal damage and provide modest support for the hypothesis that fluctuations may in part reflect altered "buffering" capacity of dopaminergic nerve terminals. However, the considerable overlap between groups suggests that other factors such as altered postsynaptic mechanisms and/or increased turnover of dopamine may make a substantial contribution to the development of motor fluctuations. PMID- 10809404 TI - Risk factors for dementia, depression and psychosis in long-standing Parkinson's disease. AB - OBJECTIVES: To study the relationships between clinical features of Parkinson's disease (PD) and the development of dementia, depression or psychosis in patients with long-standing disease. BACKGROUND: The natural history of dementia and depression in PD, and its relation to psychosis in long standing PD, are unclear. METHOD: 172 consecutive patients (99 men and 73 women, mean age at symptoms onset 58.3 +/- 13.2 years) with 5 years or more of PD (mean symptom duration of 11.8 +/ 5.6 years) were studied. Clinical data were collected during the last office visit through physical examination, detailed history, review of patient charts and outside documents. Dementia and depression were diagnosed according to DSM-IV criteria, while psychosis was diagnosed if hallucinations or delusions were present. Chi-square and t tests were used to compare the patient characteristics among those with vs. those without mental complications of the disease at different disease stages. Logistic regression was used for the comparison of associations between the presence of dementia or depression (dependent variable) and age at onset of PD, duration of PD and disease staging (explanatory variables). RESULTS: The study population consisted of 45 patients at Hoehn & Yahr (H&Y) stage < or = 2.5 (26%), 104 patients at stage 3 (60.5%) and 23 patients at H&Y stage 4-5 (13.5%). Sixty one patients (36%) had dementia, 55 patients had depression (33%) and 50 patients (27%) had psychosis. Dementia and depression were significantly associated with disease severity as reflected in the H&Y scale (P = 0.0003, Z = 3.59; P = 0.006, Z = 3.22, respectively). These associations were significant also for the older age of PD onset (> or = 59 years n = 89) subgroup (p = 0.001, Z = 3.2 for dementia and p = 0.02, Z = 2.9 for depression), but not for younger onset cases (< 59 years n = 83). Dementia was significantly associated with older age of PD onset (beta = 0.04, p = 0.009) while depression was inversely associated with age of PD onset (beta = -0.04, p = 0.02). The presence of dementia was also significantly associated with depression (beta = 1.49, p = 0.0006). Dementia and depression were found to be independent explanatory variables for the development of psychosis (logistic regression, odds ratio (OR) = 26.0, p < 0.0001; OR = 10.2, p < 0.0001, respectively). In patients with younger age of PD onset, depression more than dementia was strongly correlated with the appearance of psychosis. CONCLUSION: Dementia in PD was related to older age of symptoms onset and old age. Depression was associated with dementia or early age of PD onset. Depression seemed to contribute to the appearance of psychosis even more than dementia, especially in patients with younger age of symptoms onset. PMID- 10809405 TI - Motor performance: normative data, age dependence and handedness. AB - Quantitative, apparatus-based assessment of motor performance is a useful tool when addressing questions of therapy evaluation, quality control and early detection of movement disorders. Until now, few such methods have been sufficiently standardised to allow their routine employment in the clinic. Before assessing the impaired performance of patients, the first stage of standardisation is the determination of criteria of normal performance in healthy subjects. We have standardised a multidimensional test battery for motor performance of the upper extremities (Schoppe's "motorische Leistungs-Serie" = MLS). On the basis of the performance of 80 healthy controls, normative algorithms which allow correction of performance with reference to age were calculated. Further, our investigation provided information concerning the complexity of normal motor performance and the question of laterality (handedness). Standardised apparatus-based quantification of performance promises to be a valuable supplement to rating scales in the evaluation of patient motor skills, particularly in the early stages of their illness. PMID- 10809406 TI - Effects of apomorphine on visual functions in Parkinson's disease. AB - The effect of apomorphine on visual functions in Parkinson's disease (PD) was evaluated by use of a static contrast sensitivity test (VCTS charts), a colour discrimination test (Farnsworth-Munsell 100 Hue test) and the examination of achromatic and chromatic contour perception. 31 patients (14 male, 17 female; mean age 60.9 +/- 9.2 years) with idiopathic PD were tested before and after an individual dosage of subcutaneously applied apomorphine showing a significant effect on motor function during the whole experiment. The achromatic spatial contrast sensitivity improved significantly after apomorphine injection with respect to all spatial frequencies. The improvement of colour discrimination after apomorphine application was minimal and not statistically significant. The small advantage of apomorphine with respect to colour discrimination may be explained by negative cognitive side-effects of apomorphine interfering with the test performance. The achromatic contour perception before and after apomorphine injection was unaltered. The contour fusion latency for the green stimulus was shortened, the latency for the rest of the examined coloured stimuli was delayed (= normalized) after apomorphine application. We conclude that apomorphine may be used as a test-drug for the examination of the dopaminergic response of the visual system in patients with PD. The improvement of basal visual functions by dopaminergic stimulation with apomorphine underlines the role of dopamine deficiency for visual dysfunction in PD. PMID- 10809407 TI - The effects of the 21-aminosteroid U-74389G on spatial orientation in rats after a cerebral oligemic episode and iron-induced oxidative stress. AB - Oligemic episodes and increased iron concentration have both been proposed as being involved in neurodegenerative diseases. In animal models, a combination of both of these might therefore mimic the clinical pathology in humans. In rats, intrastriatal injections of ferric chloride, FeCl3, one week after a 60-minute oligemic episode, produced by bilateral clamping of the carotid arteries under pentobarbital anaesthesia (BCCA) impaired the animals' learning ability in a water maze task. Median adult rats, after intrastriatal 0.3 microg FeCl3, are impaired when challenged during the first three trial blocks, while after 0.06 microg FeCl3, an impairment is seen during the process of habituation to the challenge. Two-year-old animals do not show any learning effect at all after the combination of BCCA and intrastriatal FeCl3. Lazaroid U-74389G, a potent inhibitor of iron-induced lipid peroxidation, totally prevents the learning impairments in both median adult and aged animals, suggesting that iron-induced lipid peroxidation may be responsible for the late learning deficiencies. However, when U-74389G is applied one week after the oligemic episode but without the additional injection of iron, U-74389G on its own also impairs the animals' learning ability. The present animal model, when applied to clinical studies of lazaroids in humans, does seem able to give reliable information concerning the neuroprotective properties of such drugs. PMID- 10809408 TI - The activity of catechol-O-methyltransferase in parkinsonian patients with "on off fluctuations". AB - Motor fluctuations complicate therapy in Parkinson's disease (PD). The mechanisms responsible for motor fluctuations such as the on-off type, are poorly understood. Differences of activity of various enzymes, such as Catechol-O Methyltransferase (COMT, E.C.2.1.1.6.) may influence the appearance of motor fluctuations and the quantity of metabolic products of levodopa. Aim of this study was to compare erythrocyte COMT-activity in Parkinsonian patients without and with motor fluctuations from the on-off type. No significant differences in COMT-activity appeared in both groups. We conclude that activity differences in COMT-activity do not contribute to the appearance of motor fluctuations in Parkinson's disease. PMID- 10809409 TI - Tolcapone increases maximum concentration of levodopa. AB - No significant increase of the maximum concentration (Cmax) of levodopa after addition of catechol-O-methyltransferase inhibitor tolcapone occurred in previous pharmacokinetic studies predominantly on healthy volunteers. We compared pharmacokinetics of levodopa in plasma before and after addition of tolcapone in 13 treated parkinsonian subjects under standardized conditions. We found a significant increase of Cmax of levodopa after the addition of tolcapone. This may represent one cause for the occurrence of dyskinesia previously early in the course of treatment with tolcapone. PMID- 10809410 TI - Epitope analysis of myeloperoxidase (MPO) specific anti-neutrophil cytoplasmic autoantibodies (ANCA) in MPO-ANCA-associated glomerulonephritis. AB - AIM AND METHODS: Epitope analysis of sera from 20 patients with myeloperoxidase anti-neutrophil cytoplasmic antibody- (MPO-ANCA) associated glomerulonephritis was examined by Western blotting using a panel set of recombinant deletion mutants of MPO. Sera from 19 patients reacted with recombinants of MPO heavy chain, whereas no serum reacted with the light chain regions. The high frequency sites were regions on the upstream of Met341 (Ha region), on the upstream of Met409 (Hb region) near the N-terminus of the MPO heavy chain and a region on the downstream of Gly598 (Hg region) near the C-terminus. The epitope recognition profiles were classified into 2 groups. Group A, which had 1 or 2 regions in Ha, Hb and Hg, and group B, which had all 3 regions. RESULTS: Incidence of alveolar hemorrhage (AH) and pulmonary fibrosis (PF) in group A was significantly higher than that in group B (AH: group A 9 of 13 (69.2%), group B 1 of 6 (16.7%) p < 0.05, PF: group A 10 of 13 (76.9%), group B 1 of 6 (16.7%) p < 0.05, AH and/or PH: group A 12 of 13 (92.3%) and group B 1 of 6 (16.7%) p < 0.01). Relapse rate for patients in the inactive stage in group A was significantly higher than that in group B (p < 0.05). T-cell reacted regions were Ha, Hb, Hg and the light chain of MPO recombinant fragments. Higher frequency of HLA typing with MHC class II DR9 was observed. CONCLUSION: These results indicate that MPO-ANCA recognizes the linear site of the heavy chain of the MPO molecule. The epitope recognition profiles are related to the clinical features, suggesting the pathogenesis of MPO ANCA-associated glomerulonephritis. PMID- 10809412 TI - A population-based case-control teratologic study of furazidine, a nitrofuran derivative treatment during pregnancy. AB - OBJECTIVE: To study human teratogenic potential of furazidine treatment during pregnancy. DESIGN: Pair analysis of cases with congenital abnormalities and matched population controls. SETTING: The Hungarian Case-Control Surveillance of Congenital Abnormalities. PARTICIPANTS: 38,151 pregnant women who had newborn infants without any defects (population control group) and 22,865 pregnant women who had newborns or fetuses with congenital abnormalities between 1980 and 1996. RESULTS: In the case group, 157 (0.7%) and in the control group, 254 (0.7%) pregnant women were treated with furazidine. The case-control pair analysis did not indicate a teratogenic potential of furazidine use during the second to third months of gestation, i.e. in the critical period for major congenital abnormalities. CONCLUSION: Treatment with furazidine during pregnancy did not show teratogenic risk to the fetus. PMID- 10809411 TI - Platelet glycoprotein IIIa PlA1/A2 polymorphism and its relationship with diabetic nephropathy in type 2 diabetic patients. AB - OBJECTIVE: The increased prevalence of cardiovascular events in type 2-diabetic patients with micro- or macroalbuminuria is not completely explained by an excess of conventional risk factors for atherosclerosis. Genetic polymorphism within the platelet glycoprotein IIIa has been implicated in the etiology of acute coronary syndromes. We tested the hypothesis that the PI(A1/A2) polymorphism could in part account for the increased cardiovascular risk of type 2-diabetic patients with micro- or macroalbuminuria compared to normoalbuminuric diabetic patients. RESEARCH DESIGN AND METHODS: We have examined the PI(A1/A2) polymorphism of the platelet glycoprotein IIIa in type 2-diabetic patients: 94 with micro-, macroalbuminuria, and 94 with normoalbuminuria, matched for age, sex and body mass index. PI(A) genotypes were performed by polymerase chain reaction and restriction enzyme digestion. RESULTS: There was no significant difference in the prevalence of PI(A2)-positive genotypes (either PI(A1/A2) or PI(A2/A2)) in the two groups of patients (chi2 = 0.19, df = 1 , p = 0.66). CONCLUSIONS: These results suggest that carriage of the platelet glycoprotein IIIa PI(A2) allele does not contribute to explain the increased cardiovascular risk associated with micro- or macroalbuminuria in type 2 diabetes. PMID- 10809413 TI - The anion gap associated with pregnancy-induced hypertension. AB - OBJECTIVES: To determine if pregnancy-induced hypertension (PIH) is associated with a narrow anion gap. METHODS: Chart review of women with uncomplicated pregnancies and those with PIH and varying degrees of proteinuria. Electrolytes, total protein, albumin, creatinine, uric acid, a qualitative or quantitative measurement of urine protein were required for inclusion into the study. RESULTS: A low anion gap relative to women with normal pregnancies was found only in the group of PIH patients who had the greatest degree of proteinuria. The anion gap correlated directly with the concentration of plasma albumin. Women with PIH, proteinuria, and elevated liver transaminases had the lowest plasma albumin values and the smallest anion gap. CONCLUSION: The degree of hypoalbuminemia determines the magnitude of the anion gap in women with PIH and proteinuria. PMID- 10809414 TI - Prediction of colloid osmotic pressure in renal patients. AB - BACKGROUND: Colloid osmotic pressure (COP) plays a major role in transcapillary fluid shift, including in the glomerular capillary. However, COP is generally estimated by quadratic equations derived from total plasma protein and/or albumin concentrations. The aim of this study was to assess the accuracy of such equations, and to determine the potential role of liver-derived non-albumin proteins in the maintenance of COP, especially in patients presenting a nephrotic syndrome. METHODS: COP was directly assessed with an osmometer in 170 patients (347 samples), and the results compared with calculated COP, using 4 previously published formulas [Brenner 1972, Canaan-Kuhl 1993, Landis-Pappenheimer 1963, Navar 1977]. RESULTS: The 4 calculated COP values were strongly correlated with measured COP (range r = 0.88 - 0.96). However, in absolute terms, measured COP differed significantly from each of the 4 calculated mean values of COP (p < 0.001). Fibrinogen exerted per se a weak oncotic effect as measured in vitro. However, fibrinogen was highly related to albumin and presumably reflected the oncotic effect of other liver-derived non-albumin proteins. Inclusion of albumin and fibrinogen in a linear model provided an excellent fit for predicted COP with a highly significant correlation (r = 0.96, p < 0.001) over a wide range of COP values. The predicted equation was: COP(mmHg) = 6.89 x (albumin + fibrinogen) (g/dl) - 5.68. CONCLUSION: None of the 4 most commonly used formulas correctly predicted COP, and direct measurement of COP is still preferable for research studies. The introduction of fibrinogen into the formula estimating COP leads to higher accuracy, and therefore represents a more convenient model for routine evaluation. PMID- 10809415 TI - Effectiveness of erythropoiesis on supervised intradialytic oral iron and vitamin C therapy is correlated with Kt/V and patient weight. AB - BACKGROUND: Poor compliance to oral medication and diet is common in hemodialysis (HD) patients and limits the ability of oral iron therapy to support erythropoiesis. Intravenous (i.v.) iron may be associated with undesirable and sometimes life-threatening complications. PATIENTS AND METHODS: We hypothesized that intradialytic oral iron therapy can overcome compliance problems and support effective maintenance erythropoiesis, which will keep Hct in the range of 33% to 36% and EPO requirements up to 50 units/week/kg. In a prospective observational study, SC EPO-treated hospital-based HD patients without conditions known to cause EPO resistance, were managed on intradialytic oral administration of iron and vitamin C. The primary endpoints were EPO requirements and resistance to EPO which standardized EPO requirements by the Hct level. Secondary endpoints included parameters that might affect the primary endpoints. Exclusion criteria were refusal to take oral medication, prestudy Hct < 27%, recent i.v. iron therapy or transfusions, bleeding, clinical conditions obligating Hct > 30% and known causes of EPO resistance. Twelve patients completed minimal follow-up period of 9 months. RESULTS: Mean Hct was 34.4% (range: 31.8% - 40.2%). EPO requirements were 61.7 +/- 28.2 units/kg and below 52.5 units/kg in 50% of patients. Patients were classified into equal groups according to resistance to EPO, which was positively correlated (r = 0.71 p < 0.01) with body weight and Kt/V (r = -0.38, p < 0.05). CONCLUSION: In conclusion, intradialytic oral iron therapy can support effective maintenance erythropoiesis in 50% of patients without known causes for EPO resistance. High response to EPO and low EPO requirement are correlated with lower body weight and possibly improved dialysis. PMID- 10809416 TI - alpha1-microglobulin as a marker of proximal tubular damage in urinary tract infection in children. AB - The urinary alpha1-microglobulin (alpha1-M) as a marker of proximal tubular damage was measured in 86 children, aged 3/12 to 12 years, with a diagnosis of urinary tract infection (UTI) and fever. All patients had normal glomerular filtration rates (GFR). They were divided into 2 groups: A: with UTI and etiological factor E. coli, B: with UTI and etiological factor Proteus sp. Similar measurements of alpha1-M were obtained for a control group of healthy children. An increased mean level serum alpha1-M was observed in patients with UTI and fever compared to control group (p < 0.001). Urinary alpha1-M as the alpha1-microglobulin/creatinine ratio was higher in both tested group of patients with UTI and fever. Those found in group A1 and B1 before treatment were the highest and statistically significantly elevated after treatment (group A2 and B2: p < 0.001). Our results indicate the usefulness of the urinary alpha microglobulin/creatinine ratio as a marker of proximal kidney tubule damage in children with E. coli and Proteus sp. infections. Additionally, it seems to be associated with the humoral and cellular immune response. PMID- 10809417 TI - Urinary beta-glucuronidase in renal cell carcinoma. AB - The aim of the paper was to study the urinary beta-glucuronidase activity in 30 patients with renal cell carcinoma (RCC), histopathologically proven after surgery, and 32 healthy subjects taken as controls. Classification of patients was done using the TNM system. A statistically significant increase (p < 0.01) in this enzyme was found in RCC patients, without relation to the changes in the urinary sediment (hematuria, pyuria, bacteriuria) or tumor dissemination. Urinary beta-glucuronidase is a useful marker in the diagnosis of malignant renal tumors. PMID- 10809418 TI - Body composition in hemodialysis patients--is it different from that of normal subjects? AB - OBJECTIVE: Body composition assessment is an important method of evaluating nutritional and metabolic status in hemodialysis patients. PATIENTS AND METHODS: To assess the body composition of hemodialysis patients, we used dual-energy X ray absorptiometry to test 40 stable chronic hemodialysis patients and 40 normal subjects. The patients were from 38 to 70 years old, and all without diabetic mellitus. Comparing with the hemodialysis patients, the normal subjects were selected on a one-to-one base with the same sex and about the same age, body weight and height. RESULTS: The lean body mass/body weight (LBM/BW) ratio had no significant statistical difference between hemodialysis patients and normal subjects in this study (63.02 +/- 8.42% vs 64.80 +/- 7.92%, p =0.3308). The male LBM/BW ratio was higher than that of the female (71.07 +/- 4.63% vs 59.30 +/- 6.35%, p < 0.0001). According to the multiple linear regression analyses, the LBM of hemodialysis patients had positive correlation with gender (p < 0.0001), height (p = 0.0360) and weight (p < 0.0001). The total bone mineral density (BMD) of hemodialysis patients was found to be lower than that of the normal subjects (0.90 +/- 0.10 g/cm vs 0.97 +/- 0.08 g/cm, p = 0.0092). The BMD had been found to be low in the hemodialysis patients with serum intact parathyroid hormone (iPTH) > 1,700 pg/ml. The BMD had negative correlation with age in the female hemodialysis patients (r = 0.63, p = 0.0009), but no correlation in the male hemodialysis patients and in the female or male normal subjects. With the multiple linear regression analyses, the BMD of hemodialysis patients had positive correlation with weight (p = 0.0329) and negative correlation with age (p = 0.0183) and serum iPTH (p = 0.0231). CONCLUSION: We concluded that: the LBM/BW ratio of hemodialysis patients was not different from that of normal subjects. Severe secondary hyperparathyroidism hemodialysis patients had low BMD. The BMD had negative correlation with age in the female hemodialysis patients. PMID- 10809419 TI - A case of CREST syndrome and myeloperoxidase-specific anti-neutrophil cytoplasmic autoantibody-associated glomerulonephritis. AB - We report the first case of myeloperoxidase-specific anti-neutrophil cytoplasmic autoantibody (MPO-ANCA)-associated glomerulonephritis in a patient with CREST syndrome. A 74-year-old Japanese man with CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysfunction, sclerodactyly, and telangiectasia) developed rapidly progressive renal failure without elevation of blood pressure. Renal biopsy revealed glomerular sclerosis and fibrous crescents. The MPO-ANCA titer was elevated to 145 EU/ml. When patients with collagen diseases develop rapidly progressive glomerulonephritis, the possibility of MPO-ANCA-associated glomerulonephritis should be kept in mind. PMID- 10809420 TI - Chinese herb nephropathy in Japan presents adult-onset Fanconi syndrome: could different components of aristolochic acids cause a different type of Chinese herb nephropathy? AB - BACKGROUND: We encountered two cases of Chinese herb-induced Fanconi syndrome in Japan. One component of the chinese medicine was "Kan-mokutsu" (Aristolochia manshuriensis) in which aristolochic acids (AAs) were detected. METHODS: Renal biopsy showed flattening of proximal tubular epithelial cells and paucicellular interstitial fibrosis without glomerular lesions, all of which were in accordance with Chinese herb nephropathy (CHN). To date, many cases of CHN have been reported mainly as progressive renal failure in western countries. RESULTS: However, our cases were different from those in that they presented Fanconi syndrome. The detected AAs in our cases consisted of aristolochic acid (AA)-I, II and D. In contrast, in Belgium, the incriminated agent was Aristolochia fangchi which consisted of AA-I, B, C, and aristolactum. CONCLUSION: These findings could indicate that different components of AAs could cause different clinical lesions, or that the amount of ingested AAs might reflect clinical pictures, that is to say, our patients took lower volume of Chinese herbs and might be in an early stage of CHN. Furthermore, it is likely that susceptibility to this substance may be different among races. CHN would include two clinical aspects: subacute renal failure and adult-onset Fanconi syndrome. It is important to bear in mind that CHN could present Fanconi syndrome. PMID- 10809421 TI - Acquired cystic kidney disease: rapid progression from small to enlarged kidneys simulating adult polycystic kidney disease. AB - A 57-year-old man on chronic hemodialysis presented marked bilateral renal enlargement due to acquired cystic kidney disease (ACKD). He had been on hemodialysis for less than 3 years only (14 months prior to receiving a functional renal transplant which lasted 8 years, followed by 18 additional months of dialysis), before the diagnosis of ACKD was made following an episode of flank pain with gross hematuria. The marked changes in kidney appearance during this 11-year period were documented by serial ultrasound examination showing the kidneys to be of near-normal size before the start of dialysis (> or =10 cm in 1986), then shrunken and contracted 5 years later while having a functioning renal transplant (<5 cm in 1991), and markedly enlarged reaching the size of adult polycystic kidney disease after returning to dialysis (>13 cm in 1997). Since the risk of ACKD increases with duration of dialysis, we sought additional predisposing factors in this unusual case and found that 2 years after renal transplantation, the patient was diagnosed with breast cancer for which he was treated with surgical excision and tamoxifen. Based on ultrasound evidence that the tamoxifen treatment preceeded the appearance of the renal cystic changes, we wonder whether this drug may have played a role in the rapid development of ACKD. PMID- 10809422 TI - Simultaneous onset of nephrotic syndrome and insulin-dependent diabetes mellitus in a case with hypereosinophilia syndrome. PMID- 10809423 TI - Red blood cell membranes, lipid peroxidation and magnesium in renal insufficiency. PMID- 10809424 TI - Reversible proximal tubular dysfunction in a patient with acute febrile illness and normal renal function: an evidence towards leptospirosis. PMID- 10809425 TI - Dietary protein intake and nutritional status in patients with renal transplant. AB - BACKGROUND: Uremic patients on regular dialysis treatment (RDT) obtain and maintain with difficulty an adequate nutritional status. Successful kidney transplantation allows remarkable rehabilitation of patients with end-stage renal disease previously on RDT. However, information concerning the role of dietary protein restriction in the treatment of patients with chronic transplant rejection is scarce. PATIENTS AND METHODS: The role of dietary protein restriction in the treatment of patients with chronic transplant rejection was studied over 10 years in 42 patients with a kidney transplant to examine longterm renal and nutritional responses to dietary protein on graft renal function. In these patients, renal function was checked monthly, clinical evaluation and anthropometric measurements studied, nutritional status and all patients' diets were recorded. RESULTS: In 18 of these patients, biochemical signs of renal failure were found. A diet with 35 Kcal/kg and 0.7 - 0.8 grams of protein/Kg was instituted. Renal function studied every six months for 10 years showed improvement or stabilization. The low protein diet was associated with a significant reduction in 24-hour urinary protein excretion, without any change in blood pressure. Protein restriction was not associated with changes in serum protein. CONCLUSIONS: Our long-term study suggests that moderate protein intake may improve the course of chronic rejection and that restriction in protein intake may be a useful strategy in slowing the progression of renal disease in chronic rejection. PMID- 10809426 TI - Oxidative stress and nitric oxide system in post-transplant hypertension. AB - BACKGROUND: CsA-induced endothelial dysfunction and CsA-induced hypertension have been attributed to CsA effects on the endothelial-derived factors controlling vasomotor tone, but the mechanisms responsible are unclear. Endothelial nitric oxide (NO) is known to maintain a state of basal vasodilation and recently a NO mediated counterregulatory mechanism protective from CsA-induced vasoconstriction has been suggested. PATIENTS AND METHODS: Our study evaluates ecNOS gene status and NO metabolites in kidney transplanted patients under chronic CsA treatment with CsA-induced hypertension. Since CsA increases superoxide production, which metabolizes NO, plasma hydroperoxides and peroxynitrite were also evaluated as index of the presence of "oxidative stress". RESULTS: Quantification of monocyte ecNOS mRNA and NO metabolites plasma level from patients and control subjects (C) demonstrated NO system up regulation in patients notwithstanding hypertension. The mean ecNOS to beta-actin ratio was 2.00 +/- 0.87 vs 0.29 +/- 0.08 in C, p < 0.04. NO metabolite plasma level was 30.03 +/- 9.62 mM vs 9.37 +/- 3.86, p < 0.001. Hydroperoxides were also increased in patients: 3.6 +/- 1.6 i.a.u. vs 1.4 +/- 0.8, p < 0.007 (from cholesterol esters) and 10.8 +/- 6.6 vs 1.5 +/- 0.9, p < 0.008 (from triglycerides) as well as peroxynitrite plasma level: 0.36+/- 0.14 mM/L vs undetectable in C. CONCLUSIONS: This study confirms a NO system up regulation in transplanted patients. However, the counterregolatory system to CsA induced vasoconstriction, could be cancelled by CsA induced superoxide and free radicals production which, increasing NO metabolism could contribute to CsA induced vasoconstriction and hypertension. PMID- 10809427 TI - Direct effect of chronic cyclosporine treatment on collagen III mRNA expression and deposition in rat kidneys. AB - BACKGROUND: It is hypothesized that in acute and chronic CsA nephrotoxicity, in vivo models CsA side-effects are mediated by Renin-Angiotensin II (RAS)-TGF-beta 1 pathway. However, to induce chronic nephrotoxicity, CsA administration has to be combined with a low salt diet, which causes hemodynamic changes and RAS up regulation. MATERIALS AND METHODS: In order to define any direct correlation between CsA and nephrotoxicity, we studied in normal sodium fed rats, the chronic effects of CsA administration (group-1 treated with 12.5 mg/Kg/day of CsA subcutaneously; group 2 received daily placebo; group 3 interrupted CsA injection after 60 days), on renal TGF-beta-1 and collagen III expression, and on TGF-beta 1, collagen III and IV deposition. Sacrifices were performed after 2, 4, 8 and 12 weeks (wks) and kidneys were harvested for immunohistological studies and RT/PCR analysis. RESULTS: No difference of TGF-beta-1 expression and deposition was found among groups. Starting from the 2nd week of treatment, an increased collagen III deposition was evident in vessels and in outer medulla with subsequent extension at the 4th week to medullary rays and to cortex interstitium. The deposition paralleled the renal collagen III mRNA up regulation: it was significantly higher in group 1 than in group 2 (p < 0.009 at 2nd wk; p < 0.016 at 4th wk). Collagen IV deposition did not differ between groups at any point. CONCLUSIONS: Our results suggest that chronic CsA administration can induce, in normal fed rats, the process of interstitial fibrogenesis through TGF-beta non-related mechanisms. PMID- 10809428 TI - Role of antigenemia assay in the early diagnosis and treatment of CMV infection in renal transplant patients. AB - AIM: CMV antigenemia by direct pp65 antigen detection and quantification was monitored on a weekly basis during the first 3 months after kidney transplantation. SUBJECTS AND METHODS: Preemptive therapy with ganciclovir was started according to the following criteria: any positive antigemia in CMV-NEG subjects, a single determination > or = 30 cell or a two fold increase of positive cells in two consecutive specimens in CMV-POS and continued until pp65 was cleared. Overall, 109 patients were monitored. RESULTS: Among the 24 CMV-NEG patients, 13 (54%) developed a pp65 positive assay without symptoms and were treated. Ten patients remained CMV-infection free and one patient developed late onset (7 months) CMV disease (hepatitis). Among the 85 POS patients 15 (17%) developed a pp65 positive assay and were treated. Two of them developed CMV disease within 7 days of the onset of positive antigenemia and 13 were asymptomatic. The other 70 patients remained CMV-infection free. The interval between transplant and the onset of CMV infection was 39 +/- 13 days in the CMV NEG group and 64 +/- 20 days in the CMV-POS group (p < 0.001). The peak antigenemia level was 193 +/- 175 cells in the CMV-NEG group and 55+/- 78 cells in the CMV-POS group (p < 0.001). The duration of treatment did not differ in the two groups (22 +/- 7days). A second course of therapy, due to a relapse of asymptomatic infection was performed in 11/13 (85%) treated CMV-NEG patients and in 2/15 (13%) treated CMV-POS patients. CONCLUSIONS: Among the total 28 treated patients, we observed only 6 episodes of mild creatinine increase and 9 episodes of mild neutropenia. In the overall population, we observed 8 systemic infections not related to CMV. PMID- 10809429 TI - Progress in xenotransplantation. AB - BACKGROUND: XT using the pig as a donor species may provide a potential solution to the lack of human organs available for transplantation. However, two major immunological obstacles have impeded the survival of porcine organs transplanted into primates. The first is hyperacute rejection (HR), which is a consequence of the recipient's preformed antibodies binding specifically to the carbohydrate structure Gal-alpha1-3-Gal on porcine endothelial cells (EC), leading to complement activation and graft failure. Conventional strategies to overcome HR have focused on the inactivation or removal of essential components of the complement system, or removal of anti-xenograft antibodies from the blood of the recipient. Alternatively, genetically modified donor pigs, whose organs express human complement inhibitors such as human decay-accelerating factor (hDAF), have been produced. AIM: Several groups have shown that organs from these transgenic pigs do not undergo HR when transplanted into primates. Acute vascular rejection (AVR) is the second major immunological obstacle to successful XT. Several strategies are presently being investigated to overcome this form of rejection. The first approach is aimed at controlling the induced anti-xenograft humoral response with immunosuppressive agents primarily directed against lymphocytes. A second approach is aimed at preventing EC activation in the graft by inhibition of NF-kappaB, a transcription factor which is known to play a central role in EC activation. A third approach is aimed at preserving the antithrombotic properties of EC which usually disappear when these cells are activated. Finally, other groups are attempting to tackle AVR by exploring the role of platelet inhibitors or by using antibodies directed against adhesion molecules. To date, the speed and strength of the humoral xenograft rejection process have impeded in vivo studies of the cell-mediated immune response in the pig-to-primate model. However, there is now evidence that human T cells respond vigorously to pig MHC antigens and some authors have proposed that the induction of tolerance is essential if XT is to succeed clinically. It is clear that additional in vivo data still need to be generated in order to fully comprehend the involvement of the cellular immune response in this model. CONCLUSIONS: It has now been demonstrated that organs from hDAF transgenic pigs sustain the life of primates for up to 12 weeks before failing due to the onset of AVR. It is anticipated that once AVR has been overcome, long-term survival of porcine organs transplanted into primates should be reproducibly achievable. PMID- 10809430 TI - Parathyroid hormone and bone metabolism in kidney-transplanted patients. AB - BACKGROUND: Decreases in bone mass and increased susceptibility to fractures are well-recognized complications in organ transplants. SUBJECTS AND METHODS: We performed a cross-sectional study on 60 patients (40 males, 20 females, mean age 43.2 +/- 1.06, SE range 22 - 70) who underwent kidney transplantation (KTX) 55.6 +/- 4.5 months before. Blood and 24-hour urine samples were analyzed for the main parameters of mineral metabolism, and also for osteocalcin (BGP), bone alkaline phosphatase (b-ALP, urine N-telopeptid (u-NTx) and urine galactosyl-hydroxylysine (u-Ghyl). DEXA scan of the lumbar spine (LS) and proximal femur (PF) and ultrasound determination of the heel (stiffness) was also performed. RESULTS: T score values for bone density (BD) were 2.14 +/- 0.11 SD's for LS, -2.56 +/- 0.09 for PF and 2.49 +/- 0.15 for stiffness. There were 29 peripheral fractures in 16 patients. The rate of fractures before KTX were 0.0011 per patient/year and 0.0005 after transplantation (p < 0.02). When expressed as number of SD's with respect to normal controls, BGP (1.48 +/- 0.23), b-ALP (0.95 +/- 0.19), u-NTx excretion correlated negatively with BD at the femoral neck (p < 0.02) and trochanter (p < 0.03). Cumulative steroids intake were negatively correlated with b-ALP positively (p < 0.05). Current CsA was positively correlated with b-ALP (p < 0.001). Both cumulative steroid (p < 0.02) and CSA (p < 0.01) intakes were negatively correlated with BD at Wards triangle. CONCLUSIONS: Our data demonstrate an important bone depletion at each stage KTX. PTH plays a major role in the observed increase in bone turnover, exacerbating the negative effects on the bone on immunosuppressive treatment. Glucocorticosteroid therapy is an important risk factor for osteoporosis in this setting also. PMID- 10809431 TI - New immunosuppressive treatment in kidney transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) has been successfully introduced into clinical practice with evident benefits for renal transplant recipients. SUBJECTS AND METHODS: To evaluate some clinical results of MMF introduction, two groups of subjects underwent cadaveric renal transplants over the last 3 years and were retrospectively investigated. The first group (AZA group) contained 40 subjects (26 males and 14 females) on triple-drug therapy with steroids, cyclosporine and azathioprine (AZA). The second group (MMF group) contained 25 patients ( 19 males and 6 females) on the same regime with steroids and cyclosporine but MMF was administered as a third drug instead of AZA. The AZA group received renal transplant after a mean dialytic time of 32 +/- 19 months and the AZA group's dialytic time was 39.9 +/- 17 months. Clinical data, collected after a minimum 12 months observational period included a crude mortality rate and survival analysis recognized by Kaplan-Meyer curve, creatinine, creatinine clearance, rejection episodes and major clinical events such as infections and acute tubular necrosis. RESULTS: One subject died in each group. For kidney graft survival, Kaplan Meyer survival analysis showed a mean survival time of 1170.04 days in the AZA group vs 845 in the MMF group without statistical significance. Graft survival demonstrated 5:40 (12.5%) graft losses in the AZA group vs no kidney transplant loss in the MMF group (the only deceased patient had a well functioning kidney). The curve of graft cumulative proportion survival analysis demonstrated a more improved survival in the MMF group, but this difference did not reach a statistical significance (p = 0.07). Acute rejection episodes in the AZA group were 37.5% vs. 20% in the MMF group. In both groups, CMV infection was successfully treated with specific antiviral agents. CONCLUSIONS: MMF represents an important step towards induction and maintenance of immunosuppression. Our experience in a relatively small cohort investigated in a single center, demonstrates encouraging results regarding graft survival in comparison to those detected in conventional triple drug therapy. Surprisingly, in spite of stronger immunosuppressive treatment, the prevalence of CMV infections was not statistically different in the MMF versus the AZA group. PMID- 10809432 TI - Mycophenolate mofetil (MMF) in the treatment of chronic renal rejection. AB - BACKGROUND: Experimental studies suggest the efficacy of MMF in the treatment of chronic renal rejection in rats. Studies on the efficacy of MMF in chronic renal rejection in man are scarce and controversial. AIM: The aim of this study was to verify in a prospective non-randomized study the efficacy of MMF given at the dose of 2 g/day in substitution of azathioprine (AZA) in the chronic rejection of cadaveric kidney transplantation. PATIENTS AND METHODS: Twelve patients with histologically proven chronic renal rejection were enrolled. The patients were 5 males and 7 females. Mean age 38.3 +/- 13.8 years, with a mean duration of transplant of 39 +/- 19 months. Mean serum creatinine values at -6, -3, 0, +3, +6, +12 months were respectively 1.72 +/- 0.33, 1.84 +/- 0.36, 2.15 +/- 0.50, 1.88 +/- 0.54, 1.81 +/- 0.71, 1.73 +/- 0.58 mg/dl. Mean creatinine clearance values were 58.85 +/- 10.06,48.8 +/- 13.3,45.8 +/- 10.2, 54.7 +/- 13.3, 51 +/- 12.7, 57.7 +/- 18.5 ml/min. Mean deltaGFR before MMF was -2.15 ml/month. RESULTS: After MMF introduction, the overall GFR decrease attenuated. In particular in seven patients after MMF administration, we obtained a significant reduction of mean serum creatinine value (1.84 +/- 0.55 vs. 1.38 +/- 0.41mg/dl; p = 0.004). In three patients, we obtained a stabilization in GFR. Two patients were slowly progressing even after MMF introduction. After a switch to MMF in almost all patients, we obtained an improvement of renal function. In three patients, we obtained a stabilization of renal function without regression. In particular, seven patients showed a remarkable improvement of renal function. CONCLUSIONS: In conclusion our data even if concerning a small number of patients, confirm the efficacy of MMF in the treatment of renal allograft chronic dysfunction. PMID- 10809433 TI - Liver-kidney-transplantation in type 1 primary hyperoxaluria: description and comments on a case. AB - BACKGROUND: Primary hyperoxaluria leads to oxalosis, a systemic illness with fatal prognosis in uremic youngsters because of systemic complications. CASE REPORT: A 14-year old boy with primary type 1 hyperoxaluria who had a long lasting history of nephrolithiasis and passed from normal renal function to end stage renal disease within 7 months. MEASUREMENT of alanine: glyoxylate aminotransferase (AGT) catalytic activity in the liver biopsy disclosed very low activity which was not. responsive to pyridoxin., thus the patient entered onto a priority national waiting list for liver-kidney transplantation and a week later received a combined transplant. In order to increase body clearance of oxalate, the patient underwent medical treatment to increase urine oxalate solubility (sodium and potassium citrate oral therapy, magnesium supplementation and increase of diuresis) and intensive dialysis both before and after transplantation. COMMENT: The medical approach to the treatment of this rare illness is discussed. Since the major risk for the grafted kidney is related to the oxalate burden, i.e. oxalate deposition from the body deposits to the kidney that becomes irreversibly damaged, treatment consists of increasing the body clearance of oxalate both by increasing oxalate solubility in the urine and with intensive dialysis performed both before and after combined transplantation. To the same extent (by limiting body oxalate deposits), a relatively early (native GFR 20-25 ml/minute) transplantation is advisable. PMID- 10809434 TI - Kidney transplantation together with another solid organ from the same donor--a single-center progress report. AB - AIM: The aim of this study was to update our center's experience with combined renal transplants. SUBJECTS AND METHODS: Between January 1987 and March 1997, 93 segmental pancreas and kidney transplants with bladder drainage and cyclosporine A-based immunosuppression were performed followed by another 40 combined pancreatoduodenal-renal transplants with enteric drainage and FK-based immunosuppression until December 1998. After a mean observation time of 87 and 13.6 months respectively, 1-year survival for patient, kidney and pancreas was 95%, 90% and 77% and 100%, 97.5% and 87.5%, respectively. Rejection and graft thrombosis were major causes of graft loss in the bladder drainage group and peripancreatitis in the enteric drainage group. From 23 December 1983, 20 patients received a combined liver-kidney transplant. RESULTS: Main indications were glomerulonephritis and viral-induced cirrhosis. 5-year survival for patients and kidneys was 70% and for liver 62%. No rejections were identified in renal transplants, suggesting a potential immunoprotective effect of the liver. Septic complications were responsible for early death and disease recurrence for late patient loss. CONCLUSIONS: From our experience with combined kidney transplants, we conclude that simultaneous transplantation of the pancreas does not adversely affect the outcome of renal transplantation and that after combined liver-kidney transplantation, the liver appears to immunologically protect the kidney. PMID- 10809435 TI - Lamivudine for treating active hepatitis B in renal transplant recipients: a case report. AB - AIM: Chronic hepatitis B is still a matter of concern among renal transplantation patients and patients waiting for a renal transplant since it influences negatively morbidity and mortality. Morbidity and mortality are associated with HBV replication. Lamivudine is a new antiviral agent whose use has been advocated to treat HBV-infected liver transplanted patients. SUBJECT AND METHODS: Here we present our experience with an HBV-positive kidney-liver transplanted patient treated with lamivudine after transplantation. RESULTS: After lamivudine was started HBV-DNA became negative (chemiluminescence, Digene Hybrid Capture System, USA 1997) and ALT levels returned to normal. After eighteen months and after steroid pulses treatment for acute rejection, HBV-DNA became positive again, probably due to virus mutation. Lamivudine treatment was not withdrawn since it has been suggested that the mutant form might be less pathogenic than the wild one. To this extent, more than 10 months after, our patient is still in a good clinical general condition and still takes lamivudine 75 mg/day. No lamivudine related side effects were recorded. CONCLUSIONS: Our case confirms that lamivudine is a safe and useful tool in treating renal transplant recipients with chronic hepatitis B. PMID- 10809436 TI - Long-term therapy for postrenal transplant erythrocytosis with ACE inhibitors: efficacy, safety and action mechanisms. AB - SUBJECTS AND METHODS: Eleven patients with postrenal transplant erythrocytosis were treated with angiotensin-converting enzyme inhibitors for 18 months. RESULTS: Indices of red blood cell production and destruction, renal function and kalemia were followed-up during the treatment. Angiotensin-converting enzyme inhibitors (ACEi) led to a progressive and significant fall in hemoglobin and hematocrit compared to the basal values at every time interval considered with a maximum reduction at 12th month for hemoglobin ( 17.04 +/- 0.21 vs. 15.10 +/- 0.42 g/dl, p < 0.0001) and at the 6th month for hematocrit (53.74 +/- 0.56 vs. 45.7 +/- 1.19 %, p < 0.0001). Serum erythropoietin levels were reduced significantly after the first month of therapy (13.56 +/- 2.7 vs. 7.48 +/- 1.6 mUI/dl, p < 0.05) after which no further variations were recorded. In the course of ACEi therapy, there was an inverse correlation between percentage reduction in erythropoietin values and those of hemoglobin emerged (r = 0.24, p < 0.05) and hematocrit (r = 0.39, p < 0.01). Indices of red blood destruction including haptoglobin, bilirubin and lactic dehydrogenase, were unaffected by ACE inhibitors treatment, as was creatininemia and kalemia. Treatment was well tolerated by all patients who completed the study without major adverse side effects. No patient required phlebotomy and no thromboembolic event occurred during angiotensin-converting enzyme therapy. CONCLUSIONS: Our results suggest that angiotensin-converting enzyme inhibitors are a safe and effective long-term therapy of postrenal transplant erythrocytosis. These agents decrease the erythropoietin synthesis but this effect is not the sole mechanism by which angiotensin-converting enzyme inhibitors act in postrenal renal transplant erythrocytosis. PMID- 10809437 TI - Mycophenolate mofetil (MMF) versus azathioprine (AZA) in pancreas transplantation: a single-center experience. AB - AIM: Advances in immunosuppression and careful monitoring for rejection are largely responsible for improved results in pancreas transplantation. We conducted a retrospective study to establish the effectiveness of immunosuppressive therapy with mycophenolate mofetil (MMF) instead of azatioprine (AZA) in pancreas transplantation and to assess adverse effects in the two different immunosuppressive regimes. SUBJECTS AND METHODS: Since 1991, 27 pancreas transplantations were performed in 25 patients at our Institute. For induction therapy, immunosuppressant protocol consisted of quadruple immunosuppressive therapy with cyclosporine, steroids, antilymphocyte globulin and AZA in 13 patients or MMF in 12 patients respectively. RESULTS: Acute rejection occurred in 76% of patients in the AZA group compared with 53% in the MMF group. Steroid-resistant rejection was observed in 7% in the MMF group compared to 38% of patients on AZA (p < 0.01). Two kidney grafts were lost due to acute rejection in the AZA group, one pancreas was lost due to acute rejection and one to chronic rejection in the MMF group. There were no significant differences in CMV infection. Severe fungal infections were noted in 2 patients treated with MMF. Malignancy occurred in 1 patient (pancreas graft lymphoma) in MMF. CONCLUSIONS: In conclusion, patients treated with MMF required less frequent and less intensive treatment for acute rejection. However, its short- and long term side effects should be further investigated. PMID- 10809438 TI - Why should we implement living donation in renal transplantation? AB - BACKGROUND: Renal transplantation started with living donor transplants. However, after the introduction of cyclosporine, the improved results of kidney transplants from cadaveric donors have raised controversy regarding the use of living donors. There are various reasons as to why some transplant centers tend to refuse living donation: first of all, the possibility that unilateral nephrectomy can be harmful to a healthy individual. SUBJECTS AND METHODS: By reviewing the medical literature on the various aspects of living donation, postoperative mortality in connection with living donation has been calculated to be 1:3,000. RESULTS: Long-term follow-up investigations of donors demonstrated that the risk of progressive renal failure, hypertension, and proteinuria was not increased by nephrectomy per se, but other causes were responsible for that in occasional patients. From these studies, one can conclude that unilateral nephrectomy is not harmful to a healthy individual. In addition, there are other valid reasons to expand living donation: 1) the need for cadaveric donor kidneys for transplantation far exceeding the supply; 2) the better kidney quality from living donors due to the shorter ischemia time, the lack ofagonal phase and cytokines release that follows brain death; 3) the continuing improved results of kidney transplants from living donors in comparison with those from cadaveric donors in the cyclosporine era also. This appears to be true also for kidney transplants from unrelated living donors in spite of complete incompatibility with recipients. 4) Pre-emptive transplantation, based on living donors, not only avoids the risks, cost, and inconvenience of dialysis, but is also associated with better graft survival than transplantation after a period of dialysis, particularly within the live donor cohort. CONCLUSIONS: In conclusion, living donor transplants should be part of any transplant center's activity. To encourage living donation, every center should have a formal recipient family education program in conjunction with national organ donation campaigns. PMID- 10809439 TI - Attitudes and knowledge about transplantation in dialyzed patients requesting a cadaveric kidney graft. AB - AIM: Eighty-two patients answered a multiple choice questionnaire aimed at identifying their presumed and actual knowledge regarding transplantation, given immediately before evaluation by our transplant team for inclusion on our kidney transplant waiting list. SUBJECTS, METHODS AND RESULTS: A total of 78% stated that they had no or incomplete knowledge of transplantation and 22% were very well informed. The mean score for technical knowledge of transplantation (duration, requirement for removal of native kidneys, possibility of obtaining a second transplant, duration of immunosuppressive therapy and duration of the risk of rejection) was 3.1 +/- 0.15 SEM (maximal possible score 5), that for risk knowledge (risks of infections, unpleasant side effects, hypertension, diabetes mellitus, viral infections and cancer) was 1.4 +/- 0.15 (maximal possible score 6). A total of 23% knew that the spouse could donate a kidney, 74% stated that only a blood relative could and 3% that living donation was impossible. CONCLUSIONS: There is scarce knowledge about transplantation, especially with regard to the risks and living donation. PMID- 10809440 TI - The rice R gene family: two distinct subfamilies containing several miniature inverted-repeat transposable elements. AB - The R and B genes of maize regulate the anthocyanin biosynthetic pathway and constitute a small gene family whose evolution has been shaped by polyploidization and transposable element activity. To compare the evolution of regulatory genes in the distinct but related genomes of rice and maize, we previously isolated two R homologues from rice (Oryza sativa). The Ra1 gene on chromosome 4 can activate the anthocyanin pathway, whereas the Rb gene, of undetermined function, maps to chromosome 1. In this study, rice R genes have been further characterized. First, we found that an Rb cDNA can induce pigmentation in maize suspension cells. Second, another rice R homologue (Ra2) was identified that is more closely related to Ra1 than to Rb. Domesticated rice and its wild relatives harbor multiple Ra-like and Rb-like genes despite the fact that rice is a true diploid with the smallest genome of all the grass species analyzed to date. Finally, several miniature inverted-repeat transposable elements (MITEs) were found in R family members. Their possible role in hastening the divergence of R genes is discussed. PMID- 10809441 TI - A xenobiotic-stress-activated transcription factor and its cognate target genes are preferentially expressed in root tip meristems. AB - In plants, as-1-type cis elements and their trans-acting factors confer tissue specific and signal-responsive activities to the promoters of several glutathione S-transferase (GST) genes. Regulation of as-1 is widely thought to involve trans acting factors that belong to a family of basic/leucine-zipper 'TGA factors' that selectively bind this element. We have previously shown that TGA 1a, a highly conserved TGA factor of tobacco, enhances transcription through as-1 in response to xenobiotic-stress cues. To better understand the functional contribution of this transcription factor to the expression of as-1-regulated genes, we have studied its tissue- and cell-specific localization in tobacco seedlings. We show here that the relative amount of TGA1a transcripts expressed in roots and shoots correlate with the as-1-regulated, basal-level expression of a GUS transgene and two putative target GST genes. In situ hybridization of intact seedlings demonstrated that TGA1a and these GST genes are preferentially expressed in root tip meristems. Similar findings were made with a gene-specific probe for PG13, a homologue of TGA1a, demonstrating that both factors are likely to be present in the same root meristem cells. Furthermore, TGA1a protein was immunologically detected exclusively in the primary root and its meristem. Collectively, these studies suggest that TGA1a, and perhaps PG13, may contribute to the expression of GST isoenzymes, especially in root tip meristems. The biological significance of these observations is discussed. PMID- 10809442 TI - A peanut seed lipoxygenase responsive to Aspergillus colonization. AB - Several lines of evidence have indicated that lipoxygenase enzymes (LOX) and their products, especially 9S- and 13S-hydroperoxy fatty acids, could play a role in the Aspergillus/seed interaction. Both hydroperoxides exhibit sporogenic effects on Aspergillus spp. (Calvo, A., Hinze, L., Gardner, H.W. and Keller, N.P. 1999. Appl. Environ. Microbiol. 65: 3668-3673) and differentially modulate aflatoxin pathway gene transcription (Burow, G.B., Nesbitt, T.C., Dunlap, J. and Keller, N.P. 1997. Mol. Plant-Microbe Interact. 10: 380-387). To examine the role of seed LOXs at the molecular level, a peanut (Arachis hypogaea L.) seed gene, PnLOX1, was cloned and characterized. Analysis of nucleotide sequence suggests that PnLOX1 encodes a predicted 98 kDa protein highly similar in sequence and biochemical properties to soybean LOX2. The full-length PnLOX1 cDNA was subcloned into an expression vector to determine the type(s) of hydroperoxide products the enzyme produces. Analysis of the oxidation products of PnLOX1 revealed that it produced a mixture of 30% 9S-HPODE (9S-hydroperoxy-10E, 12Z-octadecadienoic acid) and 70% 13S-HPODE (13S-hydroperoxy-9Z, 11E-octadecadienoic acid) at pH 7. PnLOX1 is an organ-specific gene which is constitutively expressed in immature cotyledons but is highly induced by methyl jasmonate, wounding and Aspergillus infections in mature cotyledons. Examination of HPODE production in infected cotyledons suggests PnLOX1 expression may lead to an increase in 9S-HPODE in the seed. PMID- 10809443 TI - Organization and structural evolution of four multigene families in Arabidopsis thaliana: AtLCAD, AtLGT, AtMYST and AtHD-GL2. AB - The Arabidopsis Genome Initiative has released up to now more than 80% of the genome sequence of Arabidopsis thaliana. About 70% of the identified genes have at least one paralogue. In order to understand the biological function of individual genes, it is essential to study the structure, expression and organization of the entire multigene family. A systematic analysis of multigene families, made possible by the amount of genomic sequence data available, provides important clues for the understanding of genome evolution and plasticity. In this paper, four multigene families of A. thaliana are characterized, namely LCAD, HD-GL2, LGT and MYST. Members of HD-GL2 and LCAD have already been reported in plants. The LGT genes specify proteins containing motifs of glycosyl transferase. No plant genes similar to the LGT genes have been reported to date. The novel MYST family, most likely plant-specific, encodes proteins with no identified function. Sequencing and in silico analysis led to the characterization of 29 novel genes belonging to these four gene families. The organization, structure and evolution of all the members of the four families are discussed, as well as their chromosome location. Expression data of some of the paralogues of each family are also presented. PMID- 10809444 TI - Characterization of Ricinus communis phloem profilin, RcPRO1. AB - The mature, functional sieve tube, which forms the conduit for assimilate distribution in higher plants, is dependent upon protein import from the companion cells for maintenance of the phloem long-distance translocation system. Using antibodies raised against proteins present in the sieve-tube exudate of Ricinus communis (castor bean) seedlings, a cDNA was cloned which encoded a putative profilin, termed RcPRO1. Expression and localization studies indicated that RcPRO1 mRNA encodes a phloem profilin, with some expression occurring in epidermal, cortex, pith and xylem tissue. Purified, recombinant RcPRO1 was functionally equivalent to recombinant maize profilin ZmPRO4 in a live cell nuclear displacement assay. The apparent equilibrium dissociation constant for RcPRO1 binding to plant monomeric (G-)actin was lower than the previously characterized maize profilins. Moreover, the affinity of RcPRO1 for poly-L proline (PLP) was significantly higher than that for recombinant maize profilins. Within the sieve-tube exudate, profilin was present in 15-fold molar excess to actin. The data suggest that actin filament formation is prevented within the assimilate stream. These results are discussed in terms of the unique physiology of the phloem. PMID- 10809445 TI - Differential accumulation of Aux/IAA mRNA during seedling development and gravity response in cucumber (Cucumis sativus L.). AB - The plant hormone auxin transcriptionally activates Aux/IAA genes. We have isolated three Aux/IAA cDNA from cucumber, two cDNAs (CS-IAA1 and CS-IAA2) containing the complete open reading frame (ORF), and one partial cDNA (CS-IAA3). Northern blotting analysis showed that Aux/IAA mRNAs were induced during the emergence of radicles from seed coats. After radicle emergence, their mRNAs accumulated in the basal part of the hypocotyl much more than in the apical part, and later in elongating region of hypocotyls. CS-IAAI and CS-IAA3 mRNA significantly accumulated in response to auxin, although the increment of the former mRNA accumulation by auxin application was much greater than that of the latter. CS-IAA2 did not show an apparent change by auxin treatment in our experiment. In horizontally germinating seedlings, the transition zone between hypocotyl and root curves was due to downward gravitropic growth. On the other hand, vertically germinating seedlings of cucumber do not curve in the early stage of seedling development. The CS-IAA1 mRNA accumulation in horizontally germinating seedlings was more than that in vertically germinating ones during radicle emergence. Furthermore, asymmetric distribution of CS-IAA1 mRNA was detected in the transition zone in in situ hybridization analysis. These results suggest that the CS-IAA1 gene product may be involved in the gravity response during early development of seedlings. PMID- 10809446 TI - Cellular expression and regulation of the Medicago truncatula cytosolic glutamine synthetase genes in root nodules. AB - In this paper we have studied the localisation of expression of the two functional cytosolic glutamine synthetase (GS) genes, MtGSa and MtGSb, in root nodules of the model legume Medicago truncatula. We have used a combination of different techniques, including immunocytochemistry, in situ hybridisation and promoter beta-glucuronidase (GUS) fusions in transgenic plants, to provide the means of correlating gene expression with protein localisation. These studies revealed that transcriptional regulation (mRNA synthesis) plays an important part in controlling GS protein levels in nodules of M. truncatula. The major locations of cytosolic GS mRNA and protein are the central tissue, the parenchyma and the pericycle of the vascular bundles. These findings indicate that in nodules, GS might be involved in other physiological processes in addition to the primary assimilation of ammonia released by the bacterial nitrogenase. The two genes show different but overlapping patterns of expression with MtGSa being the major gene expressed in the infected cells of the nodule. Promoter fragments of 2.6 kb and 3.1 kb of MtGSa and MtGSb, respectively, have been sequenced and primer extension revealed that the MtGSb promoter is expressed in nodules from an additional start site that is not used in roots. Generally these fragments in the homologous transgenic system were sufficient to drive GUS expression in almost all the tissues and cell types where GS proteins and transcripts are located except that the MtGSa promoter fragment did not express GUS highly in the nodule infected cells. These results indicate that the cis-acting regulatory elements responsible for infected-cell expression are missing from the MtGSa promoter fragment. PMID- 10809447 TI - Cloning of Arabidopsis thaliana phosphatidylinositol synthase and functional expression in the yeast pis mutant. AB - It is believed that phosphatidylinositol (PI) metabolism plays a central role in signalling pathways in both animals and higher plants. PI is synthesized from CDP diacylglycerol (CDP-DG) and myo-inositol by phosphatidylinositol synthase (PI synthase, EC 2.7.8.11). Here we report the identification of a plant cDNA (AtPIS1) encoding a 26 kDa PI synthase from Arabidopsis thaliana. The plant enzyme as deduced from its cDNA sequence shares 35-41% identical amino acids with PI synthases from Saccharomyces cerevisiae and mammals. AtPIS1 functionally complements a mutant of S. cerevisiae with a lesion in PI synthase, and recombinant AtPIS1 protein present in yeast membranes strongly depends on the two principal substrates, myo-inositol and CDP-DG, and requires Mg2+ ions for full activity. PMID- 10809448 TI - Tomato contains homologues of Arabidopsis cryptochromes 1 and 2. AB - Cryptochromes are blue light photoreceptors found in both plants and animals. They probably evolved from photolyases, which are blue/UV-light-absorbing photoreceptors involved in DNA repair. In seed plants, two different cryptochrome (CRY) genes have been found in Arabidopsis and one in Sinapis, while three genes have been found in the fern Adiantum. We report the characterisation of tomato CRY genes CRY1 and CRY2. They map to chromosomes 4 and 9, respectively, show relatively constitutive expression and encode proteins of 679 and 635 amino acids, respectively. These proteins show higher similarity to their Arabidopsis counterparts than to each other, suggesting that duplication between CRY1 and CRY2 is an ancient event in the evolution of seed plants. The seed plant cryptochromes form a group distinct from the fern cryptochromes, implying that only one gene was present in the common ancestor between these two groups of plants. Most intron positions in CRY genes from plants and ferns are highly conserved. Tomato cryl and cry2 proteins carry C-terminal domains 210 and 160 amino acids long, respectively. Several conserved motifs are found in these domains, some of which are common to both types of cryptochromes, while others are cryptochrome-type-specific. PMID- 10809450 TI - Findings from SUPPORT and HELP: an introduction. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment. Hospitalized Elderly Longitudinal Project. PMID- 10809449 TI - Tobacco TGA factors differ with respect to interaction with NPR1, activation potential and DNA-binding properties. AB - In higher plants, as-1-like cis elements mediate auxin- and salicylic acid inducible transcription. Originally found in viral and T-DNA promoters, they are also functional elements of plant promoters activated during the defence response against pathogens. Tobacco bZIP transcription factor TGA1a was the first recombinant protein shown to bind to as-1. cDNAs for two novel tobacco as-1 binding bZIP proteins (TGA2.1 and TGA2.2) were isolated. Revealing a high degree of amino acid identity in the bZIP domain (89%) and the C-terminus (79%), the two TGA2 factors differ remarkably with respect to the length of the N-terminus (170 amino acids in TGA2.1 versus 43 amino acids in TGA2.2). TGA2.1 and TGA2.2, but not TGA1a, interacted with ankyrin repeat protein NPR1, a central activator of the plant defence response. In contrast, TGA2.1 and TGA1a, but not TGA2.2, functioned as transcriptional activators in yeast. Apart from conferring transcriptional activation, the N-terminal domain of TGA2.1 led to reduced in vitro as-1-binding activity and almost completely abolished binding to one half site of this bifunctional element. When being part of a heterodimer with TGA2.2, TGA2.1 was efficiently recruited to a single half site, though double occupancy of the element was still preferred. In contrast, TGA1a preferred to bind to only one palindrome, a feature that was also maintained in heterodimers between TGA1a and TGA2.1 or TGA2.2. PMID- 10809451 TI - Predicting functional status outcomes in hospitalized patients aged 80 years and older. AB - OBJECTIVE: To develop a model estimating the probability of a patient aged 80 years or older having functional limitations 2 months and 12 months after being hospitalized. DESIGN: A prospective cohort study. SETTING: Four teaching hospitals in the US. PARTICIPANTS: Enrolled patients were nonelective hospital admissions aged 80 years or older who stayed in hospital at least 48 hours. The 804 patients who survived and completed an interview at 2 months and the 450 who completed an interview at 12 months were from the 1266 patients in the Hospitalized Elderly Longitudinal Project (HELP) (76% and 47% of survivors, respectively). Median age of the 2-month survivors was 84.7 years. MEASUREMENTS AND MAIN OUTCOMES: Patient function 2 and 12 months after enrollment was defined by the number of dependencies in Activities of Daily Living (ADLs). Ordinal logistic regression models were constructed to predict functional status. Predictors included demographic characteristics, disease category, geriatric conditions, severity of physiologic imbalance, current quality of life, and exercise capacity and ADLs 2 weeks before study admission. RESULTS: Before admission, 39% of patients were functionally independent in ADLs. Of patients who survived and were interviewed at 2 months, 32% were functionally independent, and at 12 months, 36% were independent. Among patients with no baseline dependencies, 42% had developed one or more limitations 2 months later, and 41 % had limitations 12 months later. The patient's ability to perform activities of daily living at baseline was the most important predictor of functional status at both 2 and 12 months. In a multivariable predictive model, independent predictors of poorer functional status at 2 months included: worse baseline functional status and quality of life; depth of coma, if any; lower serum albumin level; presence of dementia, depression, or incontinence; being bedridden; medical record documentation of need for nursing home; and older age. Model performance, assessed using Somers' D, was 0.61 for 2 months and 0.57 for 12 months (Receiver Operating Characteristic (ROC) area = 0.81 and .79, respectively.) Bootstrap validation of the month 2 model also yielded a Somers' D = 0.60. The models were well calibrated over the entire risk range. The ROC area for prediction of the loss of independence was 0.76 for 2 months and 0.68 for 12 months. CONCLUSIONS: Many older patients are functionally impaired at the time of hospitalization, and many develop new functional limitations. A limited amount of readily available clinical information can yield satisfactory predictions of functional status 2 months after hospitalization. Models like this may prove to be useful in clinical care. This work illuminates a potential method for risk adjustment in research studies and for monitoring quality of care. PMID- 10809452 TI - Prediction of survival for older hospitalized patients: the HELP survival model. Hospitalized Elderly Longitudinal Project. AB - OBJECTIVE: To develop and validate a model estimating the survival time of hospitalized persons aged 80 years and older. DESIGN: A prospective cohort study with mortality follow-up using the National Death Index. SETTING: Four teaching hospitals in the US. PARTICIPANTS: Hospitalized patients enrolled between January 1993 and November 1994 in the Hospitalized Elderly Longitudinal Project (HELP). Patients were excluded if their length of hospital stay was 48 hours or less or if admitted electively for planned surgery. MEASUREMENTS: A log-normal model of survival time up to 711 days was developed with the following variables: patient demographics, disease category, nursing home residence, severity of physiologic imbalance, chart documentation of weight loss, current quality of life, exercise capacity, and functional status. We assessed whether model accuracy could be improved by including symptoms of depression or history of recent fall, serum albumin, physician's subjective estimate of prognosis, and physician and patient preferences for general approach to care. RESULTS: A total of 1266 patients were enrolled over a 10-month period, (median age 84.9, 61% female, 68% with one or more dependency), and 505 (40%) died during an average follow-up of more than 2 years. Important prognostic factors included the Acute Physiology Score of APACHE III collected on the third hospital day, modified Glasgow coma score, major diagnosis (ICU categories together, congestive heart failure, cancer, orthopedic, and all other), age, activities of daily living, exercise capacity, chart documentation of weight loss, and global quality of life. The Somers' Dxy for a model including these factors was 0.48 (equivalent to a receiver-operator curve (ROC) area of 0.74, suggesting good discrimination). Bootstrap estimation indicated good model validation (corrected Dxy of 0.46, ROC of 0.73). A nomogram based on this log-normal model is presented to facilitate calculation of median survival time and 10th and 90th percentile of survival time. A count of geriatric syndromes or comorbidities did not add explanatory power to the model, nor did the hospital of patient recruitment, depression, or the patient preferences for general approach to care. The physician's perception of the patient's preferences and the physician's subjective estimate of the patient's prognosis improved the estimate of survival time significantly. CONCLUSIONS: Accurate estimation of length of life for older hospitalized persons may be calculated using a limited amount of clinical information available from the medical chart plus a brief interview with the patient or surrogate. The accuracy of this model can be improved by including measures of the physician's perception of the patient's preferences for care and the physician's subjective estimate of prognosis. PMID- 10809453 TI - Age differences in care practices and outcomes for hospitalized patients with cancer. AB - OBJECTIVE: To identify age group differences in care practices and outcomes for seriously ill hospitalized patients with malignancy. DESIGN: Prospective cohort study (SUPPORT project). SETTING: Five United States teaching hospitals; data was gathered between 1989 and 1994. SUBJECTS: Nine hundred twenty five older (age > or = 65 years), 983 middle aged (age = 45-64 years), and 274 younger (age = 18-44 years) hospitalized patients receiving care for non-small cell lung cancer, colon cancer metastasized to the liver, or multi-organ system failure associated with malignancy. MEASUREMENTS: Care practices and patient outcomes were determined from hospital records. Length of survival was identified using the National Death Index. After adjusting for important variables, including severity of illness (i.e., SUPPORT model estimate for 2-month survival, cancer condition), hospital site, selection to intervention and sociodemographic variables, age group differences in care practices and outcomes were identified using general linear models. RESULTS: Older patients with cancer had lower resource utilization during hospitalization (P < .04) and were less likely to receive cancer-related treatments (i.e., chemotherapy, platelet infusions, scheduled intravenous medications) than middle-aged and young-adult patients in the first week of hospitalization (P < or = .01). More care topics were discussed with older patients and their families then with younger patients and their families (P < .001). Length of stay and total hospital costs were lower for older and middle aged patients than for younger patients. Although more older patients had discussions about transfer to hospice (P < .001), older patients were no more likely to be discharged with supportive care (inpatient hospice or home with home/ hospice care). Older patients died sooner than middle-aged patients (P < .01). CONCLUSIONS: Patient age influenced care decisions and outcomes. Older patients (age > or = 65 years) received less aggressive care, had more discussions about care decisions, and died sooner than younger patients with cancer. Younger patients had longer stays, higher hospital costs, and greater probability of rehospitalization. Although well over half of patients died within 6 months of hospitalization, few patients in any age group were discharged with supportive care. Future studies should examine age differences in palliation, as well as acute care of cancer patients across inpatient and ambulatory care settings and should assess quality of care at the end of life. PMID- 10809454 TI - The effect of nutritional supplementation on survival in seriously ill hospitalized adults: an evaluation of the SUPPORT data. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - BACKGROUND: Enteral tube and parenteral hyperalimentation are widely used nutritional support systems. Few studies examine the relation between nutritional support and patient outcomes in seriously ill hospitalized adults. OBJECTIVE: To explore the association between nutritional support and survival in seriously ill patients enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). DESIGN: A prospective study of preferences, decision-making, and outcomes. SETTING: Five teaching hospitals PARTICIPANTS: 6298 patients aged 18 or older meeting diagnostic and illness severity criteria. MEASUREMENT: Demographic characteristics, diagnoses, comorbid conditions, acute physiology score, nutritional support, and functional status before hospitalization. RESULTS: A total of 2149 patients received nutritional support. In patients who received artificial nutrition on hospital days 1 or 3 (Cohort 1), enteral feeding was associated with improved survival in coma (hazard: 0.53; 95%CI, 0.42-0.66), and reduced survival in COPD (hazard: 1.57; 95%CI, 1.18-2.08). In patients who were hospitalized on Day 7 and received artificial nutrition on days 1, 3, or 7 (Cohort 2), enteral tube feeding (hazard: 0.35; 95%CI, 0.27-0.46) or hyperalimentation (hazard: 0.58; 95%CI, 0.38-0.90) was associated with improved survival in coma. Tube feeding was associated with decreased survival in acute respiratory failure (ARF) or multiorgan system failure (MOSF) with sepsis (hazard: 1.21; 95%CI, 10.4-1.41), cirrhosis (hazard: 2.15; 95%CI, 1.35-3.42), and COPD (hazard: 1.37; 95%CI, 1.04-1.80). Hyperalimentation was associated with decreased survival in ARF or MOSF with sepsis (hazard: 1.34; 95%CI, 1.12-1.59). CONCLUSIONS: Nutritional support was associated with improved survival in coma. Enteral feeding and hyperalimentation was associated with decreased survival in ARF or MOSF with sepsis. Tube feeding was associated with decreased survival in cirrhosis and COPD. Except for patients in coma, artificial nutrition was not associated with a survival advantage. PMID- 10809455 TI - Blood transfusion administration in seriously ill patients: an evaluation of SUPPORT data. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - BACKGROUND: Administration of blood transfusion in seriously ill patients is highly variable. Limited data are available to guide transfusion decisions. OBJECTIVE: To explore characteristics of patients who received blood transfusions and decisions to forego transfusions in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). DESIGN: Prospective study of preferences, decision-making, and outcomes. SETTING: Five teaching hospitals. PARTICIPANTS: 9105 patients aged 18 years and older meeting defined diagnostic and illness severity criteria. MEASUREMENT: Data included blood transfusions, demographic characteristics, diagnoses, comorbid conditions, acute physiology score (APS), nutritional support, and functional status before hospitalization. RESULTS: A total of 2863 patients (31.4%) received blood transfusions, usually early in their hospitalization. Transfused patients were more likely male (57.3 %; P = .008), with a younger mean age (56 vs 64 years), significantly higher APS (P < .001), and significantly lower 2- and 6-month survival predictions (P < .001). The patients with acute respiratory failure or multiorgan system failure with sepsis (1714; 59.9% of all patients receiving transfusions), multiorgan system failure with malignancy (480, 16.8%), and cirrhosis (248, 8.7%) were more likely to receive blood than those with other diseases. Few patients made a decision not to receive blood before (5; 0.05%) or after (126; 1.4%) study entry. Most patients with decisions to forego transfusions also had decisions against trying resuscitation. CONCLUSIONS: Most transfusions occurred in patients with acute respiratory or multiorgan system failure. Few patients decide to forego transfusions. Additional investigation is necessary to evaluate blood transfusion practices in seriously ill patients. PMID- 10809456 TI - Physician understanding of patient resuscitation preferences: insights and clinical implications. AB - OBJECTIVE: To describe physician understanding of patient preferences concerning cardiopulmonary resuscitation (CPR) and to assess the relationship of physician understanding of patient preferences with do not resuscitate (DNR) orders and in hospital CPR. DESIGN: We evaluated physician understanding of patient CPR preference and the association of patient characteristics and physician-patient communication with physician understanding of patient CPR preferences. Among patients preferring to forego CPR, we compared attempted resuscitations and time to receive a DNR order between patients whose preference was understood or misunderstood by their physician. PATIENTS/SETTING: Seriously ill hospitalized adult patients were enrolled in the Study to Understand Prognoses and Preferences for the Outcomes of Treatments. GENERAL RESULTS: Physicians understood 86% of patient preferences for CPR, but only 46% of patient preferences to forego CPR. Younger patient age, higher physician-estimated quality of life, and higher physician prediction of 6-month survival were independently associated with both physician understanding when a patient preferred to receive CPR and physician misunderstanding when a patient preferred to forego CPR. Physicians who spoke with patients about resuscitation and had longer physician-patient relationships understood patients' preferences to forego CPR more often. Patients whose physicians understood their preference to forego CPR more often received DNR orders, received them earlier, and were significantly less likely to undergo resuscitation. CONCLUSIONS: Physicians often misunderstand seriously ill, hospitalized patients' resuscitation preferences, especially preferences to forego CPR. Factors associated with misunderstanding suggest that physicians infer patients' preferences without asking the patient. Patients who prefer to forego CPR but whose wishes are not understood by their physician may receive unwanted treatment. PMID- 10809457 TI - A prospective study of patient-physician communication about resuscitation. AB - OBJECTIVE: To evaluate prospectively seriously ill patients' characteristics, perceptions, and preferences associated with discussing resuscitation (CPR) with their physicians. DESIGN: Prospective cohort. SETTING: Five academic medical centers. PARTICIPANTS: Patients enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments who had not communicated with their physicians about CPR at admission to a hospital for life-threatening illness (n = 1288). MEASUREMENTS: Baseline surveys of patients' characteristics, health status, desires for participation in medical decision making, and cardiopulmonary resuscitation. Two month follow-up surveys of patients' communication of resuscitation preference. Chart reviews for clinical indicators. RESULTS: Thirty percent of patients communicated their resuscitation preference to their physician during a 2 month-period following hospital admission. Patients whose preference was to forego CPR (odds ratio (OR) 2.9;(95% CI, 1.9-4.2)) and whose preference had changed from desiring to foregoing CPR (OR 1.6; (95% CI, 1.1 2.4)) were more likely to communicate their preference than patients who continued to prefer to receive CPR. However, only 50% of patients who maintained a preference to forego CPR communicated this over a 2-month period. Having an advance directive and remaining in the hospital at 2-month follow-up were also independently associated with communication, whereas patients' preference for participation in decision-making, health status, and prognostic estimate were not. CONCLUSIONS: Communication about resuscitation preferences occurred infrequently after hospital admission for a serious illness, even among patients wishing to forego resuscitation. Factors such as declining quality of life, which were expected to be associated with communication, were not. An invitation to communicate about CPR preference is important after hospital admission for a serious illness. Novel approaches are needed to promote physician-patient discussions about resuscitation. PMID- 10809458 TI - Family satisfaction with end-of-life care in seriously ill hospitalized adults. AB - OBJECTIVE: To examine factors associated with family satisfaction with end-of life care in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). DESIGN: A prospective cohort study with patients randomized to either usual care or an intervention that included clinical nurse specialists to assist in symptom control and facilitation of communication and decision-making. SETTING: Five teaching hospitals in the United States. PARTICIPANTS: Family members and other surrogate respondents for 767 seriously ill hospitalized adults who died. MEASUREMENTS: Eight questionnaire items regarding satisfaction with the patient's medical care expressed as two scores, one measuring satisfaction with patient comfort and the other measuring satisfaction with communication and decision-making. RESULTS: Sixteen percent of respondents reported dissatisfaction with patient comfort and 30% reported dissatisfaction with communication and decision-making. Factors found to be significantly associated with satisfaction with communication and decision-making were hospital site, whether death occurred during the index hospitalization (adjusted odds ratio (AOR) 2.2, 95% CI, 1.3-3.9), and for patients who died following discharge, whether the patient received the SUPPORT intervention (AOR 2.0, 1.2-3.2). For satisfaction with comfort, male surrogates reported less satisfaction (0.6, 0.4-1.0), surrogates who reported patients' preferences were followed moderately to not at all had less satisfaction (0.2, 0.1-0.4), and surrogates who reported the patient's illness had greater effect on family finances had less satisfaction (0.4, 0.2-0.8). CONCLUSIONS: Satisfaction scores suggest the need for improvement in end-of-life care, especially in communication and decision making. Further research is needed to understand how factors affect satisfaction with end-of-life care. An intervention like that used in SUPPORT may help family members. PMID- 10809459 TI - Decision-making and outcomes of prolonged ICU stays in seriously ill patients. AB - BACKGROUND: Despite concern about the high costs and the uncertain benefit of prolonged treatment in the intensive care unit (ICU), there has been little research examining decision-making and outcomes for patients with prolonged ICU stays. OBJECTIVES: To evaluate decision-making and outcomes for seriously ill patients with an ICU stay of at least 14 days. DESIGN: A prospective cohort study. SETTING: Five teaching hospitals. PARTICIPANTS: Seriously ill patients enrolled in the Study To Understand Prognoses and Preferences for Risks and Outcomes of Treatments (SUPPORT). MEASUREMENT: Patients, their surrogate decision makers, and their physicians were interviewed about prognosis, communication, and goals of medical care. Based on age, diagnoses, comorbid illnesses, and acute physiology data, the SUPPORT Prognostic Model provided estimates of 6-month survival on study days 1, 3, 7, and 14. Hospital costs were estimated from hospital billing data. RESULTS: Of the 9105 patients enrolled in SUPPORT, 1494 (16%) had ICU stays of 14 days or longer. The median length of stay in an ICU was 4 days for the entire SUPPORT cohort and 35 days for patients who were treated in an ICU for 14 days or longer. Median hospital costs were $76,501 for patients who had ICU stays 14 days or longer and $10,916 for patients who did not have long ICU stays. Fifty-five percent of patients with long ICU stays had died by 6 months, and an additional 19% had substantial functional impairment. Among patients with ICU stays of at least 14 days, only 20% had estimates of 6-month survival that fell below 10% at any time during their hospitalization. For patients with long ICU stays, the mean predicted probability of 6-month survival was 0.46 on study Day 3 and 0.47 on study Day 14. Fewer than 40% of patients (or their surrogates) reported that their physicians had talked with them about their prognoses or preferences for life-sustaining treatment. Among the patients who preferred a palliative approach to care, only 29% thought that their care was consistent with that aim. Those who discussed their preferences for care with a physician were 1.9 times more likely to believe that treatment was in accord with their preferences for palliation (95% CI, 1.4-2.5) CONCLUSIONS: Prolonged ICU stays were expensive and were often followed by death or disability. Patients reported low rates of discussions with their physicians about their prognoses and preferences for life-sustaining treatments. Many preferred that care focus on palliation and believed that care was inconsistent with their preferences. Patients were more likely to receive care consistent with their preferences if they had discussed their care preferences with their physicians. PMID- 10809460 TI - Withholding versus withdrawing life-sustaining treatment: patient factors and documentation associated with dialysis decisions. AB - OBJECTIVE: We evaluated prospectively the use of acute hemodialysis among hospitalized patients to identify demographic and clinical predictors of and chart documentation concerning dialysis withheld and withdrawn. DESIGN: Prospective cohort study. SETTING: Five teaching hospitals. PATIENTS: Five hundred sixty-five seriously ill hospitalized patients who had not previously undergone dialysis who developed renal failure. MAIN OUTCOME MEASURES: Patient demographics, clinical characteristics, preferences, and prognostic estimates associated with having dialysis withheld rather than initiated and withdrawn rather than continued. Differences in chart documentation concerning decision making for dialysis withheld, withdrawn, and continued. RESULTS: Older patient age, cancer diagnosis, and male gender were associated with dialysis withheld rather than withdrawn. Age and gender differences persisted after adjustment for patients' aggressiveness of care preference. Worse 2-month prognosis was associated with both withholding and withdrawing dialysis. Chart documentation of decision-making was lacking more often for patients with dialysis withheld than for dialysis withdrawn. CONCLUSIONS: Measuring the equity of life-sustaining treatment use will require evaluation of care withheld, not just care withdrawn. Older patients and men, after accounting for prognosis and function, are more likely to have dialysis withheld than withdrawn after a trial. Further exploration is needed into this disparity and the inadequate chart documentation for patients with dialysis withheld. PMID- 10809461 TI - Patients who want their family and physician to make resuscitation decisions for them: observations from SUPPORT and HELP. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment. Hospitalized Elderly Longitudinal Project. AB - OBJECTIVE: To determine the extent to which older or seriously ill inpatients would prefer to have their family and physician make resuscitation decisions for them rather than having their own stated preferences followed if they were unable to decide themselves. DESIGN: Analysis of existing data from the Hospitalized Elderly Longitudinal Project (HELP) and the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment (SUPPORT). SETTING: Five teaching hospitals in the United States. PARTICIPANTS: 2203 seriously ill adult inpatients (SUPPORT) and 1226 older inpatients (HELP) who expressed preferences about resuscitation and about advance decision-making. MEASURES: We used a logistic regression model to determine which factors predicted preferences for family and physician decision-making. RESULTS: Of the 513 HELP patients in this analysis, 363 (70.8%) would prefer to have their family and physician make resuscitation decisions for them whereas 29.2% would prefer to have their own stated preferences followed if they were to lose decision-making capacity. Of the 646 SUPPORT patients, 504 (78.0%) would prefer to have their family and physician decide and 22.0% would prefer to have their advance preferences followed. Independent predictors of preference for family and physician decision-making included not wanting to be resuscitated and having a surrogate decision-maker. CONCLUSIONS: Most inpatients who are older or have serious illnesses would not want their stated resuscitation preferences followed if they were to lose decision-making capacity. Most patients in both groups would prefer that their family and physician make resuscitation decisions for them. These results underscore the need to understand resuscitation preferences within a broader context of patient values. PMID- 10809462 TI - Living and dying with chronic obstructive pulmonary disease. AB - OBJECTIVE: To characterize chronic obstructive pulmonary disease (COPD) over patients' last 6 months of life. STUDY DESIGN: A retrospective analysis of a prospective cohort from the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). SETTING: Hospitalization for exacerbation of COPD at five US teaching hospitals. PARTICIPANTS: COPD patients who died within 1 year (n = 416) among 1016 enrolled. METHODS: Interview and medical record data were organized into time windows beginning with death and ending 6 months earlier. OUTCOME MEASURES: Days in hospital, prognosis, illness severity, function, symptoms, patients' preferences, and impacts on families. RESULTS: One-year survival was 59%, 39% had > or = 3 comorbidities, and 15 to 25% of the patients' last 6 months were in hospitals. Exacerbation etiologies included respiratory infection (47%) and cardiac problems (30%). Better quality of life predicted longer survival (ARR: 0.36; 95% CI, 0.19-0.87) as did heart failure etiology of exacerbation (ARR: 0.57; CI, 0.40, 0.82). Estimates of survival by physicians and by prognostic model were well calibrated, although patients with the worst prognoses survived longer than predicted. Patients' estimates of prognosis were poorly calibrated. One-quarter of patients had serious pain throughout, and two-thirds had serious dyspnea. Patients' illnesses had a major impact on more than 25% of families. Patients' preferences for Do-Not Resuscitate orders increased from 40% at 3 to 6 months before death to 77% within 1 month of death; their decisions not to use mechanical ventilation increased from 12 to 31%, and their preferences for resuscitation decreased from 52 to 23%. CONCLUSIONS: Patients with advanced COPD often die within 1 year and have substantial comorbidities and symptoms. Adequate description anchors improved care. PMID- 10809463 TI - The last six months of life for patients with congestive heart failure. AB - OBJECTIVE: To characterize the experiences of patients with congestive heart failure (CHF) during their last 6 months of life. DESIGN: A retrospective analysis of data from a prospective cohort study. SETTING: Five geographically diverse tertiary care academic medical centers. PARTICIPANTS: A total of 1404 patients enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) with a diagnosis of an acute exacerbation of CHF, of whom 539 patients died within 1 year of their index hospitalization. METHODS: Data from interviews with patients or their surrogates were collected and chart abstractions performed at several time points in SUPPORT. To describe progression to death, we constructed four observational windows backward in time, beginning with patients' dates of death and ending with their date of entry into the SUPPORT project or 6 months before death, whichever came first. For each outcome, patients contributed information to all windows for which they had data collected. We describe frequency distributions for each outcome over time and report tests for trend. OUTCOME MEASURES: Outcomes examined over time included: percentage of days spent in a hospital; model-based prognostic estimates of 6-month survival; functional status; occurrence of severe physical and emotional symptoms, including pain, depression and anxiety; patients' preferences for care; and the financial impact of patients' illnesses on their families. RESULTS: As death approached, patients' prognoses became poorer and illnesses more severe. Median Acute Physiology Scores for hospitalized patients rose from 33 in the interval 6 months to 3 months before death, to 44 within 3 days of death. However, the median model-based estimate of 6-month survival was 54% even within 3 days of death. Number of functional impairments, median depression scores and percent of patients reporting severe pain or dyspnea increased as death approached, with 41% of patient surrogates reporting that the patient was in severe pain and 63% reporting that the patient was severely short of breath during the 3 days before death. Perceived quality of life did not change appreciably, with 29 to 58 % of patients reporting good to excellent quality of life in all intervals before death. As death approached, patients were more likely to prefer Do Not Resuscitate (DNR) status, with the percent of patients preferring DNR rising from 33% at 6 months to 3 months before death to 47% at 1 month to 3 days before death (P < .05). The frequency with which DNR orders were written for hospitalized patients also increased as death approached. The patients' illnesses had marked financial impact on their families, with 23 % of patients' families reporting the loss of most or all of family savings at the time of the patient's death. CONCLUSIONS: As death approaches during the last 6 months of life in CHF, illness becomes more severe, disability and the experience of certain symptoms more frequent, and patient preference not to be resuscitated more common. However, there is no significant decrement in quality of life as death approaches. Reflecting the unpredictable course of CHF during the last month of life, many patients have good median model-based 6-month prognoses and enjoy good to excellent quality of life. PMID- 10809464 TI - Dying with cancer: patients' function, symptoms, and care preferences as death approaches. AB - OBJECTIVE: To characterize the dying experience of patients with cancer over the last 6 months of life. STUDY DESIGN: A retrospective analysis of the last 6 months of life among patients with colon cancer and non-small cell lung cancer enrolled in a prospective cohort study from June 1989 to June 1991 and from January 1992 to January 1994. SETTING: Five geographically diverse tertiary care academic medical centers participating in the Study to Understand Patient Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) Project. PARTICIPANTS: All patients enrolled in SUPPORT who had either colon cancer, metastatic to the liver or stage III or stage IV non-small cell lung cancer and died within 1 year of their index hospitalization. This report examines 316 of 520 patients with metastatic colon cancer and 747 of 939 patients with lung cancer enrolled in SUPPORT. METHODS: Data were collected by interview and chart abstraction at several time points in the SUPPORT Project. To describe progression to death, we constructed four observational windows backward in time beginning with patients' date of death and ending with their date of entry into the SUPPORT Project or 6 months before their death, whichever came first: (1) 3 days before death, (2) 3 days to 1 month before death, (3) 1 month to 3 months before death, and (4) 3 months to 6 months before death. For each outcome, patients contributed information to all windows during which they had data collected. We describe the frequency distributions of each outcome over time and report tests for trend. OUTCOME MEASURES: We examined several outcomes over time, including: percentage of days spent in a hospital; prognosis as measured by model based prognostic estimates of 6-month survival; severity of illness as measured by the acute physiology score; functional status as measured by dependencies in activities of daily living (ADLs); severe physical and emotional symptoms, including pain, depression, and anxiety; patients' preferences for care; and the financial impact on patients' families. RESULTS: The death rate within 1 year of study entry was high among patients with metastatic colon cancer and advanced non small cell lung cancer enrolled in SUPPORT (61% and 80%, respectively). As patients with cancer progress toward death, their estimated 6-month prognosis decreases significantly and the severity of their disease worsens. Patients' functional status also declines significantly as they approach death, such that most patients have four or more impairments within the 3 days before death. Patients with cancer experience significantly more pain and confusion as death approaches. Severe pain is common; more than one-quarter of patients with cancer experience serious pain 3 to 6 months before death and more than 40% were in serious pain during their last 3 days of life. However, dying patients are only modestly depressed and anxious during their last 3 days of life. As death approaches, patients favor comfort measures over life-extension, and about two thirds want to forego resuscitation within 3 days of death. Families of patients dying with cancer incurred significant financial burdens during the last 6 months of life, and much of this burden was already experienced by 3 to 6 months before death. CONCLUSIONS: The last 6 months of life for patients with cancer is characterized by functional decline and poorly controlled severe pain and confusion. Although patients increasingly prefer comfort care as they near death, many die in severe pain. These findings highlight important opportunities to improve the quality of care at the end of life for patients dying with cancer. PMID- 10809465 TI - Dying with end stage liver disease with cirrhosis: insights from SUPPORT. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment. AB - OBJECTIVES: To understand patterns of care and end-of-life preferences for patients dying with end stage liver disease with cirrhosis (ESLDC). METHODS: Data were collected during the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment (SUPPORT), a prospective cohort study of seriously ill hospitalized adults at five teaching hospitals in the United States, and included all patients enrolled in SUPPORT with ESLDC. RESULTS: Of 575 patients with ESLDC, 166 died during index hospitalization, and 168 died in the following year. The majority were male (65%) and white (80%); the median age was 52 years. Most rated their quality of life as poor or fair, and multiple comorbidities were common. Most spent their last few days completely disabled. Families often reported loss of most income and the need to leave work or other activities in order to care for patients. Pain was at least moderately severe most of the time in one-third of patients. End-of-life preferences were not associated with survival. Most patients (66.8%) preferred CPR, but DNR orders and orders against ventilator use increased near death. CONCLUSIONS: Patients with liver disease were young, likely to be male, and often had low incomes. The high burden of pain was comparable to that reported for patients with lung and colon cancer. Persons dying with liver disease may benefit from increased attention to relief of symptoms, improved home care, and advanced care planning. PMID- 10809466 TI - Elderly persons' last six months of life: findings from the Hospitalized Elderly Longitudinal Project. AB - BACKGROUND: Few studies describe the end of life in very old people. OBJECTIVES: To characterize the last 6 months of life and dying in patients 80 years and older by describing demographic characteristics, functional state and quality of life, symptoms, preferences, use of life-sustaining treatments, satisfaction with care, and family burden. DESIGN: A retrospective analysis for patients enrolled in a prospective cohort study. SETTING: Four teaching hospitals who participated in the Hospitalized Elderly Longitudinal Project (HELP). SUBJECTS: 417 patients who died within 1 year of their enrollment hospitalization. MEASUREMENTS: Chart reviews and interview data with patients and surrogates at several points in time. We constructed four observational time windows backward in time beginning with the patients' death. RESULTS: Before hospitalization, two out of three patients reported fair quality of life, and patients averaged 2.4 impairments in activities of daily living. Seventy percent preferred comfort care on the third day of hospitalization. During the last month of life, three of five patients interviewed in the hospital and four of five interviewed out of the hospital preferred not to be resuscitated. At the time of death, four of five patients had a do not resuscitate (DNR) order and two of five had an order to withhold a ventilator. During the last month of life, one out of four patients reported severe pain. CONCLUSIONS: Patients reported increasing functional impairments and limited quality of life. The majority preferred comfort care. The number of patients in severe pain was substantial. Before death, the majority had measures in place to limit aggressive care. PMID- 10809467 TI - Dying with acute respiratory failure or multiple organ system failure with sepsis. AB - BACKGROUND: The dying experience of patients with acute respiratory failure (ARF) or multiple organ system failure with sepsis (MOSF) has not been described. OBJECTIVES: To describe patients dying from ARF or MOSF, including demographic characteristics, baseline function and quality of life, symptoms, preferences, use of life-sustaining treatments, satisfaction with care, and family burden. DESIGN: A multicenter prospective study. SETTING: Five US teaching hospitals, in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). PARTICIPANTS: A total of 1295 adults who died during hospitalization for ARF or MOSF. MEASUREMENTS: Chart reviews and interviews with patients and surrogates. RESULTS: SUPPORT enrolled 2956 patients with ARF or MOSF, and 44% died during enrollment hospitalization. Quality of life before hospitalization was reported as fair by 87% of patients. The mean number of impairments in their baseline activities of daily living was 1.6. At the time of death, 79% had a DNR order and 31% had an order to withhold ventilator support. The average time from the DNR order to death was 2 days. Dying patients spent an average of 9 days on a ventilator. Surrogates indicated that one out of four patients died with severe pain and one out of three with severe confusion. Families of 42% of the patients who died reported one or more substantial burden. CONCLUSIONS: Patients in this study reported substantial functional impairments and reduced quality of life. Limitations to aggressive treatments were usually implemented only when death was imminent. Family impact and physical and emotional suffering were substantial. PMID- 10809468 TI - Dying with lung cancer or chronic obstructive pulmonary disease: insights from SUPPORT. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - CONTEXT: Many are calling for patients with advanced chronic obstructive pulmonary disease (COPD) to receive hospice care, but the traditional hospice model may be insufficient. OBJECTIVE: To compare the course of illness and patterns of care for patients with non-small cell lung cancer and severe COPD. DESIGN: Prospective cohort study of seriously ill, hospitalized adults. SETTING: Five teaching hospitals in the United States. PATIENTS: Patients with Stage III or IV non-small cell lung cancer (n = 939) or acute exacerbation of severe COPD (n = 1008). MAIN OUTCOME MEASURES: Patients' preferences for pattern of care and for ventilator use; symptoms; life-sustaining interventions; and survival prognoses. RESULTS: Sixty percent in each group wanted comfort-focused care; 81% with lung cancer and 78% with COPD were extremely unwilling to have mechanical ventilation indefinitely. Severe dyspnea occurred in 32% of patients with lung cancer and 56% of patients with COPD and severe pain in 28 % of patients with lung cancer and 21% of patients with COPD. Patients with COPD who died during index hospitalization were more likely than patients with lung cancer to receive mechanical ventilation (70.4% vs 19.8%), tube feeding (38.7% vs 18.5%), and cardiopulmonary resuscitation (25.2% vs 7.8%). Mechanical ventilation had greater short term effectiveness in patients with COPD, based on survival to hospital discharge (76% vs 38%). Patients with COPD maintained higher median 2-month and 6 month survival prognoses, even days before death. CONCLUSIONS: Hospitalized patients with lung cancer or COPD preferred comfort-focused care, yet dyspnea and pain were problematic in both groups. Patients with COPD were more often treated with life-sustaining interventions, and short-term effectiveness was comparatively better than in patients with lung cancer. In caring for patients with severe COPD, consideration should be given to implementing palliative treatments more aggressively, even while remaining open to provision of life sustaining interventions. PMID- 10809469 TI - Description of the SUPPORT intervention. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - BACKGROUND: The purpose of Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) was to improve outcomes for seriously ill hospitalized adults by improving information and decision-making. The SUPPORT intervention has been characterized only briefly in previous publications. OBJECTIVE: To characterize the intervention in SUPPORT and its implementation. DESIGN: Reports derived from training and administrative materials, quantitative descriptions of implementation activities, and qualitative analysis of narrative reports and focus group participation by the intervention nurses. SETTING AND PATIENTS: SUPPORT enrolled 2652 patients in the intervention arm and 2152 in the control arm of a block-randomized trial of enhanced information, counseling, and support. The patients were hospitalized with one of nine serious illnesses in one of five US teaching hospitals between 1992 and 1994. MEASUREMENTS: (1) Reports on training and supervisory materials; (2) Rates of intervention component completion from contact logs and reports completed by the intervention nurses; and (3) grounded theory analysis of patient narratives, overview questionnaires, and focus group transcripts from the intervention nurses. RESULTS: Prognostic reports were delivered on time to the caregiving team in 83% of cases on Day 3. Reports of surrogate interviews of patient preferences and understanding were delivered on time to the caregiving team in 72% of first week cases. Patients' own reports of preferences were unavailable for 56% of cases in the first week. Overall, 39% of the rest of the patients had their interview information delivered on time to the caregiving team. The SUPPORT intervention nurses averaged 8.5 contacts with patients, 7.6 with surrogates, 3.5 with physicians, and 11.7 with other staff. The intervention nurses felt that they were fully involved in 81% of cases and had a limited role in another 14%. The major issues for patients were: understanding their situation, communication and decision making, advance planning, do not resuscitate orders, and general support, including support for loss and grieving. The narrative sources showed that the nurses were enthusiastic, dedicated, and strong in their support of the study objectives. They identified various barriers to effectiveness and voiced doubt that the analytic targets would show an effect from the intervention. CONCLUSIONS: The SUPPORT intervention was implemented vigorously and completely. PMID- 10809470 TI - Length of stay and therapeutic intervention allow estimation of in-hospital resource use independent of site and inflation. AB - OBJECTIVES: To develop and evaluate a model to estimate inpatient resource utilization for seriously ill patients using periodic medical record abstraction and hospital length of stay. DESIGN: A prospective cohort study of prognoses and preferences and outcomes of seriously ill hospitalized adults. The predictive model was based on the Therapeutic Intervention Scoring System (TISS), length of stay (LOS), and hospital site, and was developed on Phase I data and validated on Phase II data. SETTING: Five teaching hospitals in the United States PARTICIPANTS: The derivation sample included 4301 patients enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment (SUPPORT). The validation sample included 4804 patients. MEASUREMENTS: Hospital charges were correlated with the product of hospital length of stay and average score from the TISS. Model fit assessment used R2 for total hospital charges. RESULTS: The correlation between hospital charges and the final model, after adjusting for site differences, was 0.86. The correlation was similar when validating the model in an independent data set. CONCLUSIONS: We present a method to estimate hospital resource utilization that is independent of site and inflation and can estimate actual costs in multiyear, multisite research endeavors. PMID- 10809471 TI - Survey methods for seriously ill hospitalized adults: practical lessons from SUPPORT. The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - OBJECTIVE: To describe issues confronted in collecting questionnaire-based data in a large study of seriously ill hospitalized adults, discuss methods used to achieve acceptable results, summarize the reliability of selected measures, and explore challenges for patient-based research in this population. DESIGN: A prospective cohort study. SETTING: Five teaching hospitals in the US. PARTICIPANTS: 9105 patients, with a subset of 80 patients and 10 proxy respondents for reliability testing. MEASUREMENTS AND MAIN OUTCOMES: Response rates; percent agreement and Kappa statistics to assess test-retest reliability. RESULTS: A pretest and pilot test were used to help develop the assessment strategy, establish study procedures, and identify staff characteristics likely to facilitate the project. Achieving adequate response rates required great perseverance and flexibility by interview staff as well as close monitoring and central supervision. Standardized quality assurance with timely feedback, and social support provided by peer and supervising staff were both important. Older women proved to be the best interviewers. Reliability was acceptable for existing and newly developed scales to measure subjective health status, preferences, and satisfaction. CONCLUSIONS: It was possible to collect reliable and useful data on preferences and subjective health outcomes from a cohort of seriously ill hospitalized patients and their proxies. Achieving acceptable data quality required a large interview team, mature staff, flexibility about modes of administration, and centralized, structured quality assurance. Successful outcomes research in this population is likely to require similar planning and efforts. PMID- 10809472 TI - Age-related differences in care preferences, treatment decisions, and clinical outcomes of seriously ill hospitalized adults: lessons from SUPPORT. AB - OBJECTIVES: To review previously published findings about how patient age influenced patterns of care for seriously ill patients enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). DESIGN: An observational prospective study. SETTING: Five acute care hospitals. PARTICIPANTS: A total of 9105 seriously ill patients enrolled in SUPPORT. MEASUREMENTS: The outcomes examined included patients' preferences for aggressive care, decision making regarding cardiopulmonary resuscitation and use of other life-sustaining treatments, hospital costs, intensity of resource use, and survival. RESULTS: Although older patients preferred less aggressive care than younger patients, many older patients wanted cardiopulmonary resuscitation and care focused on life extension. Patients' families and healthcare providers underestimated older patients' desire for aggressive care. After adjustment for illness severity, comorbidity, baseline function, and patients' preferences for aggressive care, older age was associated with lower hospital costs and resource intensity and higher rates of decisions to withhold life-sustaining treatments. In adjusted analyses, older age was associated with a slight survival disadvantage. This survival disadvantage persisted, even after adjustment for aggressiveness of care, suggesting that the relation between age and survival is not accounted for by less aggressive treatment of older patients. CONCLUSIONS: Even after adjustment for patients' prognoses and care preferences, seriously ill hospitalized older patients were treated less aggressively than younger patients. SUPPORT cannot fully identify whether the relationship between older age and less aggressive treatment is better explained by the withholding of potentially beneficial treatments from older patients, or by the excessive provision of ineffective treatment to younger patients. However, the latter explanation is favored by the SUPPORT finding that less aggressive treatment for older patients does not contribute to the modest survival disadvantage associated with older age. PMID- 10809473 TI - Pain and suffering in seriously ill hospitalized patients. AB - BACKGROUND: Previous studies had suggested a high prevalence of pain in hospitalized patients but had not specifically evaluated pain and other symptoms in seriously ill and older hospitalized patients. OBJECTIVE: The SUPPORT and HELP studies were designed to (1) assess the frequency and severity of pain and other symptoms during hospitalization 2 and 6 months later, and before death; (2) identify factors associated with pain and other symptoms; and (3) test an intervention to improve pain. DESIGN: An observational cohort and randomized controlled trial. SETTING: Five major teaching hospitals in the US. PATIENTS: Hospitalized patients aged 80 years and older or with one of nine serious illnesses. INTERVENTION: Education of patients and family members about pain control, monitoring of patients' pain, and feedback about pain with treatment suggestions to nurses and physicians. MEASUREMENTS: Data from the medical record and interview-based information about pain and other symptoms and preferences for care and symptom control from patients and family members. RESULTS: Pain and other symptoms were frequent and often severe in seriously ill and older patients during hospitalization, at follow-up, and before death, even in those with diseases not traditionally associated with pain. There was wide variation in symptom experience across hospitals. Patients' preference for pain control was not associated with symptom experience. The intervention did not improve pain control. CONCLUSIONS: Control of pain and other symptoms remains an important medical and ethical issue. Routine monitoring of pain and other symptoms should be linked to treatment strategies aimed at combinations of symptoms and tested to assuage concerns about side effects. PMID- 10809474 TI - Communication and decision-making in seriously ill patients: findings of the SUPPORT project. The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - OBJECTIVES: The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) represents one of the largest and most comprehensive efforts to describe patient preferences in seriously ill patients, and to evaluate how effectively patient preferences are communicated. Our objective was to review findings from SUPPORT describing the communication of seriously ill patients' preferences for end-of-life care. METHODS: We identified published reports from SUPPORT describing patient preferences and the communication of those preferences. We abstracted findings that addressed each of the following questions: What patient characteristics predict patient preferences for end of life care? How well do physicians, nurses, and surrogates understand their patients' preferences, and what variables are correlated with this understanding? Does increasing the documentation of existing advance directives result in care more consistent with patients' preferences? RESULTS: Patients who are older, have cancer, are women, believe their prognoses are poor, and are more dependent in ADL function are less likely to want CPR. However, there is considerable variability and geographic variation in these preferences. Physician, nurse, and surrogate understanding of their patient's preferences is only moderately better than chance. Most patients do not discuss their preferences with their physicians, and only about half of patients who do not wish to receive CPR receive DNR orders. Factors other than the patients' preferences and prognoses, including the patient's age, the physician's specialty, and the geographic site of care were strong determinants of whether DNR orders were written. In SUPPORT patients, there was no evidence that increasing the rates of documentation of advance directives results in care that is more consistent with patients' preferences. CONCLUSIONS: SUPPORT documents that physicians and surrogates are often unaware of seriously ill patients' preferences. The care provided to patients is often not consistent with their preferences and is often associated with factors other than preferences or prognoses. Improving these deficiencies in end-of-life care may require systematic change rather than simple interventions. PMID- 10809475 TI - The effects of patient race on outcomes in seriously ill patients in SUPPORT: an overview of economic impact, medical intervention, and end-of-life decisions. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - BACKGROUND: Black Americans have significantly lower life expectancy than white Americans. Racial differences in medical access, management, and DNR orders have been documented. OBJECTIVE: To review the effects of patient race on intervention and end-of-life decisions in seriously ill patients in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). DESIGN: Review of published analyses from SUPPORT. SETTING: Five teaching hospitals PARTICIPANTS: A total of 9105 patients aged 18 years or older (15% black race) meeting diagnostic and illness severity criteria. MEASUREMENT: Analysis of data collected by chart abstraction and interviews. RESULTS: Blacks reported significant loss in savings, although adjusting for diagnosis and disease severity did not demonstrate significant racial differences. Economic hardship was associated with a preference for comfort care, except in black patients (OR 0.71; CI 95%, 0.57-0.88). Blacks received less intervention with no significant difference in survival. Pain level and control were not affected by race. Blacks were more likely to want CPR, although adjustment for self-pay or Medicaid eliminated racial differences. Blacks were more likely to continue to prefer CPR 2 months after hospitalization (28% vs 17%) and were more likely to change a DNR order to preferring CPR (40 vs 27%). Blacks also more frequently wished to discuss CPR preferences with their physicians but were less likely to have this type of communication (OR 1.53; CI 95%, 1.11-2.11). CONCLUSIONS: Patient race may impact on medical intervention and preferences in seriously ill patients. However, in this population, the differences are of modest clinical importance. PMID- 10809476 TI - Insights about dying from the SUPPORT project. The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - OBJECTIVES: The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) project was a study of 9105 seriously ill patients, 4274 of whom died within 6 months. HELP, the Hospitalized Elderly Longitudinal Project, was an ancillary study, in four of the five SUPPORT hospitals, of 1286 persons aged 80 years and older, 321 of whom died within 6 months. This paper reviews the SUPPORT and HELP literature to bring together insights concerning the time near death of seriously ill patients. METHODS: We reviewed published reports from SUPPORT and HELP, specifically, demographics of dying; characteristics of a prognostic model to estimate survival; patient symptoms near death; patient preferences and decision-making near death; ineffectiveness of the SUPPORT intervention; costs of dying while seriously ill; and the impact of serious illness on the family. We also compared and contrasted the experience of patients with different conditions. RESULTS: Patients in SUPPORT who died were typically younger than age 75. Most SUPPORT patients who died were male and most had an income of less than $11,000, although the older patients in HELP were even more likely to have had such low incomes. Patients with cirrhosis were much younger than most decedents, and patients with cancer were less often poor. Most had serious symptoms close to death. The place of death was more closely related to hospital bed supply than to decisions made by healthcare providers or individual patient preferences or characteristics. Prognosis near death was quite uncertain, especially in patients with heart and lung failure. CONCLUSIONS: Although SUPPORT aimed to describe and compare decision-making affecting seriously ill patients, it also illuminated many other aspects of their course near death. PMID- 10809477 TI - Ineffectiveness of the SUPPORT intervention: review of explanations. AB - BACKGROUND: The aim of the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments -- SUPPORT -- was to improve the care of seriously ill patients by improving decision-making for patients with life threatening illnesses. Several theories have been proposed to explain why the SUPPORT intervention was unsuccessful at improving outcomes. OBJECTIVE: To review and discuss explanations offered by others regarding why the SUPPORT intervention failed to have a discernible impact on its prespecified outcome measures. DESIGN: A descriptive review of published articles and book chapters, with synthesis of data-based and conceptual insights. METHODS: The Medline, Bioethicsline, and Ethx databases were searched for citations to SUPPORT articles between 1994 and the end of 1998. This search was supplemented by other published materials that had come to the authors' attention. RESULTS: The critiques and explanations regarding the reasons the SUPPORT intervention did not improve outcomes were catalogued and organized into 11 major categories, the first 10 of which are explored in the present study: (1) the inception cohort was biased against an effect of the intervention, (2) the intervention was not implemented as designed, (3) the intervention failed because nurses were too readily ignored, (4) the intervention was too polite, (5) the intervention presented information ineffectively, (6) the intervention did not focus on primary care physicians, (7) the intervention falsely dichotomized do not resuscitate (DNR) decisions, (8) the intervention needed more years on site or an earlier start with each patient, (9) the intervention required more appropriate outcome measures, (10) the intervention was irrelevant because usual care is not seriously flawed, (11) the conceptual model behind SUPPORT was fundamentally flawed in aiming to improve individual, patient-level decision-making as the way to improve seriously ill, hospitalized patients' experiences. CONCLUSIONS: Although some of the critiques were found to raise important concerns, we conclude in each case that the explanation offered is inadequate to explain the failure of the intervention. We urge further reflection on the fundamental assumptions that informed the design of that intervention and refer the reader to a more comprehensive treatment of that issue in the companion paper in this volume. PMID- 10809478 TI - Rethinking fundamental assumptions: SUPPORT's implications for future reform. Study to Understand Prognoses and Preferences and Risks of Treatment. AB - BACKGROUND: The intervention in SUPPORT, the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments, was ineffective in changing communication, decision-making, and treatment patterns despite evidence that counseling and information were delivered as planned. The previous paper in this volume shows that modest alterations in the intervention design probably did not explain the lack of substantial effects. OBJECTIVE: To explore the possibility that improved individual, patient-level decision-making is not the most effective strategy for improving end-of-life care and that improving routine practices may be more effective. DESIGN: This paper reflects our efforts to synthesize findings from SUPPORT and other sources in order to explore our conceptual models, their consistency with the data, and their leverage for change. RESULTS: Many of the assumptions underlying the model of improved decision-making are problematic. Furthermore, the results of SUPPORT suggest that implementing an effective intervention based on a normative model of shared decision-making can be quite difficult. Practice patterns and social expectations may be strong influences in shaping patients' courses of care. Innovations in system function, such as quality improvement or changing the financing incentives, may offer more powerful avenues for reform. CONCLUSIONS: SUPPORT's intervention may have failed to have an impact because strong psychological and social forces underlie present practices. System-level innovation and quality improvement in routine care may offer more powerful opportunities for improvement. PMID- 10809479 TI - The collected reports from SUPPORT and HELP: an annotated bibliography of manuscripts in print as of December 31, 1999, and SUPPORT prognostic formulas. Study to Understand Prognoses and Preferences and Risks of Treatment. Hospitalized Elderly Longitudinal Project. PMID- 10809480 TI - Central nervous system hemangioblastomas, endolymphatic sac tumors, and von Hippel-Lindau disease. AB - Von Hippel-Lindau disease (VHL) is a hereditary cancer syndrome caused by germline mutations of the VHL tumor suppressor gene. Major progress has been made in the last decade in both clinical and fundamental aspects of VHL. The VHL gene product, pVHL, has major and multiple functions: pVHL regulates not only first angiogenesis but also extracellular matrix formation and the cell cycle. A molecular diagnosis of VHL is now available, leading to a transformation in clinical management of patients and their families. Diagnosis of VHL has to be suspected in patients with a VHL-related tumor without familial history and especially in case of hemangioblastoma or endolymphatic sac tumors. Such patients should be systematically investigated for clinical and molecular evidence of VHL disease. Treatment of symptomatic hemangioblastomas remains mainly neurosurgical, often in emergency, but stereotactic radiosurgery is emerging as an alternative therapeutic procedure. In the future, antiangiogenic drugs could represent a potential medical treatment of CNS hemangioblastomas in view of their highly vascular structure. Lastly, visceral manifestations of VHL disease are also of critical importance and require early detection for effective treatment. PMID- 10809481 TI - von Hippel-Lindau disease PMID- 10809482 TI - Molecular, oncologic, and therapeutic spectrum of von Hippel-Lindau disease PMID- 10809483 TI - Results in the operative treatment of elderly patients with intracranial meningioma. AB - With life expectancy in the industrial nations increasing during recent years, the number of patients older than 70 years with intracranial tumours and, especially, meningiomas is rising. To evaluate the indications for operative treatment, we reviewed 66 patients older than 70 years who were operated upon for intracranial meningioma in our department between 1991 and 1997. The mean age was 75 years. The oldest patients were 86 years old. Thirteen patients with recurrent meningiomas were operated upon. The mortality rate was 7.6%. Neurological symptoms improved in 38 patients (57.6%), were unchanged in 11 (16.6%), and deteriorated in 12 (18.2%). Patients with recurrent meningiomas seem to have a higher operative risk and their outcome is worse than after a primary operation. In general, there were good postoperative results in patients with few concomitant diseases, small meningiomas, small edema, short time of operation, and accessible location (convexity rather than skull base). Age in general is not a contraindication for operation. In cases of incidental findings of small meningiomas, we recommend observation and MRI follow-up. Symptomatic meningiomas should be removed whenever there is an acceptable risk from an internal or anaesthesiological point of view. PMID- 10809484 TI - Bilateral epidural hematoma. AB - Bilateral epidural hematomas are very rare and are associated with high mortality. The purpose of this study is to identify the clinical features, mechanisms, and outcomes of bilateral epidural hematomas. This report considers 19 cases of bilateral epidural hematoma hospitalized between 1987 and 1997. All of the cases, with the exception of three, were diagnosed within the first 6 h. The neurologic evaluations on admission and during hospital stay were based on the Glasgow Coma Scale. Hematomas were determined by CT scans in all cases. The patients were evaluated using the Glasgow Outcome Scale after 6 months. In 13 patients, the bilateral epidural hematoma was in the midline. In six patients, hematomas were at different locations on either side. Surgical approach was chosen as the primary treatment modality in 18 patients. One was treated conservatively. The mortality rate was 15.7% in this series. With the widespread use of CT scan, diagnosis before deterioration of the neurological status affects the results of surgery and prognosis or even presents the possibility of a conservative treatment. PMID- 10809485 TI - Transcranial Doppler sonography in adult hydrocephalic patients. AB - Transcranial Doppler sonography (TCD) is a noninvasive technique for measurement of cerebral blood flow velocity (CBFV) in the major arteries of the circle of Willis. Dynamic changes in the pulsatility index (PI) and the resistance index (RI), as calculated from TCD data, allow for an assessment of the forces acting on the terminal vasculature of the brain. The present study was designed to investigate a possible relationship between TCD parameters and intracranial pressure (ICP) changes in adult patients with hypertensive hydrocephalus. Blood flow velocity in the middle cerebral artery (MCA) was studied by TCD in 29 hydrocephalus patients and in 20 healthy controls. ICP was measured in the patient group before ventricular shunting and was correlated with TCD data. The mean CBFV in hydrocephalic patients prior to ventriculoperitoneal shunting was significantly lower than in the control group. Compared to normal persons, systolic and end-diastolic CBFV values in patients were significantly decreased, suggesting an increased distal cerebrovascular resistance. PI and RI values in patients with elevated ICP prior to shunting were significantly increased in comparison to those of normal persons. There was a statistically significant positive correlation of preshunting ICP and mean preshunting values of RI (r=0.50, P<0.01) in hydrocephalic patients, but no significant correlation between PI and ICP, and between CBFV and ICP. Immediately after shunting, ICP returned to normal, and PI and RI values decreased significantly, while the mean CBFV increased. In a subgroup of hydrocephalic patients with a preshunting ICP value >35 mm Hg (n=6), the changes described above were more pronounced than in the subgroup with preshunting ICP values <35 mm Hg, which suggests an exponential degree of influence of ICP on TCD parameters. In conclusion, TCD may provide a tool for assessment of ICP in adult patients with occlusive hydrocephalus, although an exact noninvasive measurement of ICP by TCD seems impossible. Changes in the RI and PI indices appear to be useful indicators of elevated ICP. PMID- 10809486 TI - Expression of cadherin and CSF dissemination in malignant astrocytic tumors. AB - Cadherins are Ca2+-dependent cell adhesion molecules that play an important role in tissue construction and morphogenesis in multicellular organisms. Because in recent years there have been reports of cadherin involvement in tumor metastasis, we conducted an immunostain for E-cadherin and N-cadherin monoclonal antibodies in paraffin-embedded surgical specimens of primary and recurrent lesions in 13 cases of glioblastoma and nine cases of anaplastic astrocytoma. No expression of E-cadherin was detected in the tumor cells. On the other hand, expression of N cadherin was observed in malignant astrocytic tumor cells, but the N-cadherin positive rate tended to be less at the time of recurrence. Decreased expression of N-cadherin was detected at the time of recurrence in 11 of the 13 cases in the glioblastoma group. Cerebrospinal fluid (CSF ) dissemination and extracranial metastasis were observed in nine (81.8%) of these 11 patients. Therefore, we tried to analyze the clinical backgrounds and the N-cadherin positive rates by statistics. We concluded that decreased expression of N-cadherin at the time of recurrence correlates with dissemination in malignant astrocytic tumors. PMID- 10809487 TI - Anatomical study of the lateral wall of the cavernous sinus, emphasizing dural construction and neural relations. AB - This study aims to determine the microscopic anatomy of the layers of the lateral wall of the cavernous sinus (CS) and, in particular, intends to examine the location and relations of the dural openings on the deep layer. Forty sides of 20 formalin-fixed and fresh cadavers were dissected and their CS examined. In 12 cases we found an opening on the deep dural layer; however, in four of them the inferolateral trunk of the internal carotid artery (ICA) was identified through these dural openings. We noticed the trochlear nerve making a curve (5% of cases) or lying close to the ophthalmic nerve (12.5%) on the lateral wall. In one case, the triangular area described by Parkinson could not be exposed surgically. Our findings indicate the importance of the heterogeneous courses of the cranial nerves lying on the lateral wall and point to the significance of the dural openings, which can influence the etiology of neoplastic invasions originating from the CS. PMID- 10809488 TI - Detection of epileptogenic focal cortical dysplasia by depth, not subdural electrodes. AB - Centers that perform presurgical epilepsy evaluations disagree on whether depth or subdural electrodes represent the optimal technique for invasive recording, especially in seizures originating outside the temporal lobe. A 13-year-old girl with a normal magnetic resonance imaging scan had unlocalized partial onset seizures, despite scalp and subdural grid ictal video/EEG recordings. Repeat video/EEG with depth electrodes showed a discrete site of continuous interictal spiking and seizure onset that was located 2-2.5 cm beneath the surface of the sensory cortex. The resected region showed focal cortical dysplasia and the patient had greater than 95% seizure frequency reduction at 3-year follow-up. We conclude that although subdural electrodes have many advantages when recording seizures outside the temporal lobes, depth electrodes may provide superior recordings when the epileptogenic region is beneath the cortical surface. PMID- 10809489 TI - Bypass surgery using a radial artery graft for bilateral extracranial carotid arteries occlusion. AB - A patient presenting with recurrent ischemic attacks was demonstrated to have complete occlusion of the right common and left internal carotid arteries. An external carotid angiogram showed a large left superficial temporal artery (STA) supplying both sides of the scalp. 123I-IMP single photon emission computed tomography (SPECT) revealed hypoperfusion of the both hemispheres, especially the left cerebral hemisphere. An extracranial-intracranial (EC-IC) bypass was performed using a radial artery graft interpositioned between the proximal part of the STA and the M2 segment, thus preserving blood flow to the scalp through the STA. Postoperative angiography after 1 year showed good circulation through the anastomosis, and 123I-IMP SPECT studies demonstrated increased cerebral perfusion. The patient improved clinically. The surgical technique is described below. PMID- 10809490 TI - Prevalence and significance of exit block during arrhythmias arising in pulmonary veins. AB - INTRODUCTION: Recent studies described the occurrence of conduction block within pulmonary veins. The purpose of this study was to evaluate the prevalence of exit block during arrhythmias that arise in pulmonary veins. METHODS AND RESULTS: Twenty-five patients with atrial tachycardia/fibrillation underwent successful ablation of 28 arrhythmogenic foci within a pulmonary vein. The prevalence of exit block in the pulmonary veins was determined in 28 arrhythmogenic pulmonary veins and 40 nonarrhythmogenic pulmonary veins. During isolated premature depolarizations, exit block in a pulmonary vein was observed at 50% of arrhythmogenic pulmonary vein sites and was never observed within pulmonary veins that did not generate a tachycardia (P < 0.01). During tachycardia, exit block from a pulmonary vein was observed in 61% of the arrhythmogenic pulmonary veins. The mean cycle length of the pulmonary vein tachycardias associated with exit block was significantly shorter than the cycle length of tachycardias that were not associated with exit block (163 +/- 32 vs 251 +/- 45 msec, P < 0.001). Exit block in two pulmonary veins during the same episode of tachycardia was observed in 3 of the 28 arrhythmogenic pulmonary veins (11%) in three different patients. Simultaneous recordings in the two pulmonary veins demonstrated bursts of tachycardia in both veins that were not synchronized. Radiofrequency catheter ablation of the arrhythmogenic site in one of the pulmonary veins eliminated spontaneous recurrences of tachycardia from the other pulmonary vein. CONCLUSION: Exit block from pulmonary veins is a common observation during tachycardias generated within pulmonary veins and indicates that an arrhythmogenic pulmonary vein has been identified. The occurrence of exit block in more than one pulmonary vein most likely is attributable to simultaneous tachycardias, one or both of which may be tachycardia induced and perpetuated by the other. PMID- 10809491 TI - Typical atrial flutter ablation: conduction across the posterior region of the inferior vena cava orifice may mimic unidirectional isthmus block. AB - INTRODUCTION: The aim of this study was to map the low right atrium before and after radiofrequency ablation of the inferior vena cava-tricuspid annulus (IVC TA) isthmus in patients with typical atrial flutter (AFI) to better understand the electrophysiologic meaning of incomplete or unidirectional block following the ablation procedure and its relationship with AFI recurrence. METHODS AND RESULTS: We performed atrial mapping in 12 patients using a "basket" catheter in the IVC orifice, Halo catheter in the right atrium, and multipolar catheters in the coronary sinus (CS) and His region. In patients in sinus rhythm, atrial activation was analyzed during pacing from the CS and low lateral right atrium (LLRA) before and after ablation. Atrial activation propagated across the isthmus and posterior region of the IVC orifice simultaneously before ablation. Mapping during AF1 in four patients showed that the crista terminalis was a site of functional block. After ablation, evaluation of Halo catheter recordings in three patients showed apparent unidirectional counterclockwise block, whereas analysis of basket catheter recordings demonstrated complete bidirectional block. The apparent conduction over the isthmus during pacing from proximal CS was due to conduction along the posterior part of the IVC orifice, which activated the LLRA despite complete isthmus block. CONCLUSION: Our results demonstrate that limited endocardial mapping may yield a pattern compatible with unidirectional block in the IVC-TA isthmus, although bidirectional block is present at this anatomic level. PMID- 10809492 TI - Body surface distribution and response to drugs of ST segment elevation in Brugada syndrome: clinical implication of eighty-seven-lead body surface potential mapping and its application to twelve-lead electrocardiograms. AB - INTRODUCTION: Body surface distribution and magnitude of ST segment elevation and their reflection in 12-lead ECGs have not been clarified in Brugada syndrome. METHODS AND RESULTS: Eighty-seven-lead body surface potential mapping and 12-lead ECGs were recorded simultaneously in 25 patients with Brugada syndrome and 40 control patients. The amplitude of the ST segment 20 msec after the end of QRS (ST20) was measured from all 87 leads, and an ST isopotential map was constructed. The maximum ST elevation (maxST20) was distributed in an area of the right ventricular outflow tract in all Brugada patients, and it was larger than that in control patients (0.37 +/- 0.13 vs 0.12 +/- 0.04 mV; P < 0.0005). The maximum was observed on the level of the parasternal fourth intercostal space, on which the V1 and V2 leads of the standard 12-lead ECG were located, in 18 of the 25 Brugada patients in whom typical coved- or saddleback-type ST elevation was seen in leads V1 and V2. The maximum was located on the second intercostal space in the remaining seven Brugada patients in whom only a mild saddleback-type ST elevation was seen in leads V1 and V2 of the 12-lead ECG. Typical ST segment elevation was recognized in leads V1 and V2, which were recorded on the second or third intercostal space. ST elevation in Brugada patients was dramatically normalized by isoproterenol, a beta-adrenergic agonist (maxST20 = 0.17 +/- 0.08 mV; P < 0.0005 vs control conditions), and accentuated by disopyramide, an Na+ channel blocker (maxST20 = 0.50 +/- 0.15 mV; P < 0.0005 vs control conditions), without any change in the location of the maxST20. CONCLUSION: Our data indicate that recordings of leads V1-V3 of the 12-lead ECG on the parasternal second or third intercostal space would be helpful in diagnosing suspected patients with Brugada syndrome. The data suggest that Na+ channel blockers are capable of accentuating ST elevation in leads V1-V3. PMID- 10809493 TI - Junctional rhythm during slow pathway radiofrequency ablation in patients with atrioventricular nodal reentrant tachycardia: beat-to-beat analysis and its prognostic value in relation to electrophysiologic and anatomic parameters. AB - INTRODUCTION: Junctional rhythm usually is considered a sensitive but nonspecific marker of successful ablation of the slow pathway in AV nodal reentrant tachycardia. Nevertheless, this junctional rhythm has been little studied, and its relations to recognized predictors of successful radiofrequency (RF) application were never established in any study. METHODS AND RESULTS: Thirty-nine patients underwent RF ablation of the slow pathway for AV nodal reentrant tachycardia. Ninety RF applications were delivered, and each ablation site was determined using three different fluoroscopic projections. Six anatomic zones were defined from low posterior septum to the site of distal His-bundle recording (P1, P2, M1, M2, A1, and A2). Characteristics of junctional rhythm during RF applications were analyzed. Atrial electrogram characteristics at the ablation sites also were studied. All patients had successful slow pathway ablation, without any complication. The ablation sites were located as follows: 41 at P1, 26 at P2, 20 at M1, and 3 in M2. Forty RF applications were successful: 14 of 41 attempts at P1, 7 of 26 at P2, 16 of 20 at M1, and 3 of 3 at M2. Mid-septal ablation site (M1 and M2) was associated with higher occurrence of junctional rhythm (P < 0.0001), earlier first junctional beat (P = 0.008), and earlier occurrence of the longest junctional burst (P = 0.03) compared with posterior ablation site (P1 and P2). The combination of a mid-septal ablation site and a first junctional beat occurring < or = 3 seconds after onset of RF application identified successful RF application with 100% accuracy. Using multivariate analysis, the ablation site, duration of atrial electrogram (including slow pathway potential when present), and occurrence of junctional rhythm were independent predictors of success. CONCLUSION: Successful slow pathway ablation depends on many factors. Junctional rhythm characteristics are related to the site of RF delivery and can be helpful in assessing successful slow pathway ablation. PMID- 10809494 TI - Late fields of the magnetocardiographic QRS complex as indicators of propensity to sustained ventricular tachycardia after myocardial infarction. AB - INTRODUCTION: Magnetocardiographic (MCG) mapping is a new method to record cardiac signals. This study examined the association of MCG late fields with the propensity to sustained ventricular tachycardia (VT) after myocardial infarction (MI). METHODS AND RESULTS: One hundred patients with remote MI were studied, 38 with and 62 without history of VT. High-resolution MCG and signal-averaged ECG (SAECG) as a comparative method were recorded. Time-domain parameters describing the abnormal low-amplitude end QRS activity, MCG late fields, and SAECG late potentials were analyzed. Late field parameters differed significantly between the patient groups: filtered QRS duration was 137 +/- 26 msec in the VT group and 110 +/- 18 msec in the control group (P < 0.001), and root mean square amplitude of the last 40 msec was 260 +/- 170 and 510 +/- 360 fT (P < 0.001), respectively. The optimal MCG parameter combination yielded a sensitivity of 92% and a specificity of 61% in classification to the VT group, whereas those for SAECG were 63% and 66%. In a subgroup of 63 patients with marked left ventricular dysfunction and comparable stage of coronary heart disease, only MCG (sensitivity 73%, specificity 67%) but not SAECG could assign a patient to the VT group. CONCLUSION: Late fields of the MCG QRS complex indicate propensity to life threatening arrhythmias in post-MI patients. This discriminative ability persists in the presence of severe left ventricular dysfunction where ECG late potentials lose their informative value. MCG late field analysis is a potential new method for noninvasive risk assessment in post-MI patients. PMID- 10809495 TI - High uniformity of left and right ventricular repolarization dynamics induced by an abrupt decrease in pacing cycle length in a dog is not affected by left ventricular ischemia. AB - INTRODUCTION: After an abrupt increase in heart rate, action potential duration (APD) will shorten. To assess the effect of ischemia on APD shortening dynamics, we compared right ventricular (RV) and left ventricular (LV) APD shortening induced by an abrupt decrease in pacing cycle length (PCL) during control and LV ischemia. METHODS AND RESULTS: In eight anesthetized AV block dogs, endocardial LV and RV APD were determined simultaneously after an abrupt PCL decrease from 800 to 350 msec. Measurements were repeated during left anterior descending coronary artery (LAD) occlusion. During control, LV and RV APD shortened 97 +/- 27 and 71 +/- 14 msec, respectively (P < 0.05). Shortening was pronounced in a short initial phase and gradual in the longer secondary phase. Linear regression analysis revealed very high uniformity of LV and RV APD shortening dynamics (r2 = 0.96 +/- 0.01). During repeated LAD occlusion, ischemia induced a gradual LV APD shortening from 314 +/- 25 msec to a new steady-state value of 251 +/- 23 msec, whereas RV APD remained stable at 289 +/- 28 msec. The additional PCL decrease resulted in LV and RV APD shortening of 72 +/- 8 and 68 +/- 15 msec, respectively, with the same high uniformity of shortening dynamics as seen during control (r2 = 0.94 +/- 0.03). CONCLUSION: There is a pronounced difference in APD shortening dynamics induced by an abrupt decrease in PCL compared with ischemia. LV shortening dynamics induced by a decrease in PCL are not affected by LV ischemia, preserving a high interventricular uniformity of repolarization dynamics. PMID- 10809496 TI - Ischemia, heart rate, and ventricular repolarization. PMID- 10809497 TI - Specific up-regulation of mitochondrial F0F1-ATPase activity after short episodes of atrial fibrillation in sheep. AB - INTRODUCTION: Ventricular fibrillation induced by either digitalis intoxication or electrical stimulation is reported to alter myocardial energy by impairing the sarcolemmal Na,K-ATPase or the receptor for digitalis and the mitochondrial ATPase synthase or F0F1-ATPase. However, little is known about these membrane functions during atrial fibrillation (AF). METHODS AND RESULTS: We analyzed the effects of electrically induced AF on biochemical activities of atrial F0F1 ATPase and Na,K-ATPase in sheep. A group of six sheep was subjected to direct short electrical stimulation of the right atrium to induce AF. Sham-operated sheep served as a control group. Microsomal and mitochondrial membranes of atrial muscle were isolated and tested for enzymatic activity. All paced sheep developed multiple episodes of sustained AF, with a mean total duration of 110 minutes over a 2-hour period. Data showed that short-term pacing-induced AF significantly activated membrane F0F1-ATPase activity (P < 0.05) without changes in cytochrome c oxidase activity, Na,K-ATPase activity, ouabain sensitivity, and alpha1-subunit expression. CONCLUSION: Specific activation of F0F1-ATPase activity is an early molecular consequence of sustained AF in sheep. PMID- 10809498 TI - Inhibition of Na,K-ATPase by external electrical cardioversion in a sheep model of atrial fibrillation. AB - INTRODUCTION: Electrical external cardioversion commonly used to treat atrial fibrillation (AF) is associated with myocardial membrane damage and disturbances in ionic homeostasis (hemodynamically unstable). The present study was designed to investigate whether alterations in ionic homeostasis observed were due in part to changes in the myocardial activity of Na,K-ATPase. METHODS AND RESULTS: AF was induced by pacing in ten anesthetized sheep divided into two groups. Group I (n = 4) received a single external countershock of 360 J after three episodes of AF lasting 10 minutes. Group II (n = 6) served as controls. Activity, responsiveness to ouabain, and membrane expression of catalytic alpha and beta subunits of Na,K ATPase in sarcolemmal myocardial membrane fractions were investigated. Membrane fluidity and fatty acid composition, and plasma levels of atrial natriuretic factor (ANF) also were measured. One shock after episodes of AF significantly decreased ventricular Na,K-ATPase activity up to 50% (P < 0.001) without modification of atrial activity at the membrane level. Sites with low affinity to ouabain showed a fivefold lower affinity for ouabain in the cardioversion group than in the control group (IC50 = 7.9 micromol/L vs 40 micromol/L ouabain, P < 0.05). Plasma levels of ANF were significantly increased in the cardioversion group compared with the control group. These changes were independent of membrane modulation in terms of expression of Na,K-ATPase, membrane fluidity, and fatty acid composition. CONCLUSION: This study suggests that left ventricular perturbation of ionic homeostasis subsequent to transthoracic cardioversion could result from inactivation of Na,K-ATPase activity. PMID- 10809499 TI - Relationship of specific electrogram characteristics during sinus rhythm and ventricular pacing determined by adaptive template matching to the location of functional reentrant circuits that cause ventricular tachycardia in the infarcted canine heart. AB - INTRODUCTION: It would be advantageous, for ablation therapy, to localize reentrant circuits causing ventricular tachycardia by quantifying electrograms obtained during sinus rhythm (SR) or ventricular pacing (VP). In this study, adaptive template matching (ATM) was used to localize reentrant circuits by measuring dynamic electrogram shape using SR and VP data. METHODS AND RESULTS: Four days after coronary occlusion, reentrant ventricular tachycardia was induced in the epicardial border zone of canine hearts by programmed electrical stimulation. Activation maps of circuits were constructed using electrograms recorded from a multichannel array to ascertain block line location. Electrogram recordings obtained during SR/VP then were used for ATM analysis. A template electrogram was matched with electrograms on subsequent cycles by weighting amplitude, vertical shift, duration, and phase lag for optimal overlap. Sites of largest cycle-to-cycle variance in the optimal ATM weights were found to be adjacent to block lines bounding the central isthmus during reentry (mean 61.1% during SR; 63.9% during VP). The distance between the mean center of mass of the ten highest ATM variance peaks and the narrowest isthmus width was determined. For all VP data, the center of mass resided in the isthmus region occurring during reentry. CONCLUSION: ATM high variance measured from SR/VP data localizes functional block lines forming during reentry. The center of mass of the high variance peaks localizes the narrowest width of the isthmus. Therefore, ATM methodology may guide ablation catheter position without resorting to reentry induction. PMID- 10809500 TI - Block of I(Ks) does not induce early afterdepolarization activity but promotes beta-adrenergic agonist-induced delayed afterdepolarization activity. AB - INTRODUCTION: An early afterdepolarization (EAD)-induced triggered beat is thought to precipitate torsade de pointes (TdP) in the long QT syndrome (LQTS). Previous studies demonstrated the development of EAD activity and dispersion of repolarization under LQT2 (reduced I(Kr)) and LQT3 (augmented late I(Na)), but not LQT1 (reduced I(Ks)), conditions. The present study examines these electrophysiologic characteristics during I(Ks) block. METHODS AND RESULTS: Canine epicardial (Epi), M, and endocardial (Endo) tissues and Purkinje fibers isolated from the canine left ventricle were studied using standard microelectrode recording techniques. The I(Ks) blocker chromanol 293B (293B, 30 microM), produced a homogeneous rate-independent prolongation of action potential duration (APD) in Epi, M, and Endo, but little to no APD prolongation in Purkinje. Chromanol 293B 1 to 30 microM failed to induce EADs or delayed afterdepolarizations (DADs) in any of the four tissue types. Isoproterenol (ISO, 0.1 to 1.0 microM) in the presence of 293B 30 microM significantly prolonged the APD of the M cell (basic cycle length > or = 1 sec), abbreviated that of Purkinje, and caused little change in that of Epi and Endo. The combination of 293B 30 microM and ISO 0.2 microM did not induce EADs in any of the four tissue types, but produced DAD activity in 4 of 8 Epi, 7 of 10 M cells, and 3 of 8 Endo. CONCLUSION: Our results indicate that I(Ks) block alone or in combination with beta-adrenergic stimulation does not induce EADs in any of the four canine ventricular tissue types, but that the combination of the two induces DADs as well as accentuated dispersion of repolarization. PMID- 10809501 TI - Catheter ablation of mitral isthmus ventricular tachycardia using electroanatomically guided linear lesions. AB - Mitral isthmus ventricular tachycardia uses a reentrant circuit with a critical isthmus of conduction bounded by the mitral valve proximally and a remote inferior infarction scar distally. Successful catheter ablation requires placement of a lesion to transect the isthmus so as to prevent wavefront propagation. We report a case with previously unsuccessful ablation in which focal isthmus ablation failed to eliminate arrhythmia. Electroanatomic mapping demonstrated a wide tachycardia isthmus, and a linear lesion placed from the edge of the inferior infarct (as demonstrated on the three-dimensional voltage electroanatomic map) to the base of the mitral valve successfully eliminated tachycardia. In some patients with mitral isthmus VT, a wide isthmus requires linear lesion placement to fully transect the isthmus and eliminate tachycardia. Electroanatomic mapping can be used to define isthmus boundaries and thus guide successful ablation. PMID- 10809502 TI - Ventricular tachycardia as a complication of atrial flutter ablation. AB - A 61-year-old woman with dilated cardiomyopathy, who previously underwent successful radiofrequency catheter ablation for atrial flutter, developed monomorphic ventricular tachycardia (VT). The site of VT origin was the inferobasal right ventricle adjacent to the previous atrial isthmus ablation area. The most likely mechanism for the VT was scar-related reentry, the scar being the result of previous radiofrequency lesions in the atrial isthmus. The VT was successfully ablated. PMID- 10809503 TI - Noncontact mapping-guided catheter ablation of atrial fibrillation associated with left atrial ectopy. AB - We report the use of a novel noncontact mapping system used to perform left atrial mapping and to guide radiofrequency ablation in two patients, each with atrial fibrillation (AF) triggered by left atrial ectopy. A noncontact multielectrode probe and ablation catheter were advanced into the left atrium through a transseptal puncture or a patent foramen ovale. Isopotential mapping delineated the focal origin at the ostium of the right lower pulmonary vein in one patient and close to the ostium of the left upper pulmonary vein in the other patient. The ablation catheter was guided to the target sites using a locator signal. The foci were ablated successfully in both patients. No recurrences of AF were observed during follow-up at 4 and 6 months, respectively. PMID- 10809504 TI - Exit block of focal repetitive activity in the superior vena cava masquerading as a high right atrial tachycardia. AB - An unusual case of atrial tachycardia (AT) originating from the superior vena cava (SVC) is reported. A 34-year-old man without structural heart disease underwent catheter ablation for drug-resistant AT. During the tachycardia, low amplitude spiky electrograms with a cycle length of 120 to 175 msec were recorded in the SVC and exhibited 2:1 exit block to the atria, masquerading as the atrial activation observed with high right AT. These spiky electrograms also were observed during sinus rhythm, but they appeared immediately after the local atrial electrograms. The spikes were traced to a point 3 cm above the junction of the right atrium. Radiofrequency ablation at the site of the earliest appearance of the spike in the SVC successfully eliminated the tachycardia. During the following 15 months, no clinically significant atrial arrhythmias, including atrial fibrillation, occurred. This report indicates that careful mapping, including inside the SVC, will be a requisite in patients with high right atrial tachyarrhythmias. PMID- 10809505 TI - Helical representation of atrial reentry: a teaching aid for electrophysiology. AB - Cardiac clinical electrophysiology is difficult to teach because it requires mental integration of complex information on cardiac activation, which includes tridimensional spatial orientation of the cardiac structures involved and variables such as refractory period, conduction velocity, and cycle length. Commonly used representations of cardiac arrhythmias include snapshot sequences or ladder diagrams. The former lack time dimension, and the latter lack spatial dimension. We propose a schematic tridimensional representation of reentry as a helicoidal shape that is the result of adding a time dimension to a circular representation. This presentation, which also can be called a "loop diagram," allows better integration of spatial phenomena with recorded electrogram sequences and can help the teaching of basic clinical electrophysiology. PMID- 10809506 TI - Gap junction remodeling in infarction: does it play a role in arrhythmogenesis? PMID- 10809507 TI - Adenosine-induced tachycardia: what is the mechanism? PMID- 10809508 TI - Noncompaction of ventricular myocardium and arrhythmias. PMID- 10809509 TI - Epicardial mapping and radiofrequency catheter ablation of ischemic ventricular tachycardia using the CARTO system. PMID- 10809510 TI - Rapacuronium. Introduction. PMID- 10809511 TI - Neuromuscular blocking drugs for the new millennium: current practice, future trends--comparative pharmacology of neuromuscular blocking drugs. PMID- 10809512 TI - Recovery from neuromuscular block and its assessment. PMID- 10809513 TI - Neuromuscular blocking drugs in pediatric patients. AB - The most useful qualities of a NMBD for pediatric anesthesia are: rapid, reliable onset of laryngeal muscle block after IV or IM administration, duration of < or =20 min, and lack of side effects. Until recently, no nondepolarizer met all these criteria. However, 2 mg/kg rapacuronium produces rapid laryngeal block that can be easily reversed to restore neuromuscular function within 20 min in most pediatric patients. PMID- 10809514 TI - The pharmacoeconomics of neuromuscular blocking drugs. PMID- 10809515 TI - Can succinylcholine be abandoned? AB - The elective use of succinylcholine in anesthesia has largely been abandoned because of unwanted side effects. Alternatives now exist for short, intermediate, or long elective surgical procedures. NMBDs are frequently used only to facilitate tracheal intubation; rapacuronium fills an important niche particularly for a short elective case (e.g., same-day surgery). However, an equally critical issue is whether there is a reliable replacement for succinylcholine for the treatment of laryngospasm or for rapid sequence induction in patients with "full stomachs." Succinylcholine produces more intense block in a shorter time at the laryngeal muscles, compared with the adductor pollicis, compared with vecuronium, rocuronium, mivacurium, and rapacuronium (30). Although most intubations can be facilitated with 80%-90% neuromuscular block, the ideal relaxant for a rapid sequence induction should produce uniformly complete neuromuscular blockade in 1 min. Variability in the degree of neuromuscular blockade and onset time can be compared for various relaxants by using the standard deviation (Table 1), the coefficient of variation (Table 2), or a plot of the degree of maximum neuromuscular block and the time to maximum block. Figure 1 shows such a plot for mivacurium (13). There is less variability in the maximum block at the larger dose of rapacuronium but still variability in onset time. Further studies will be important in defining the role of rapacuronium for rapid sequence induction in various clinical settings. PMID- 10809516 TI - The role of recovering physicians in 19th century addiction medicine: an organizational case study. AB - An elaborate network of inebriate homes, inebriate asylums, nationally franchised private addiction treatment institutes, and proprietary home cures for addiction arose on the American landscape between 1850 and 1900. The pinnacle of the movement to professionalize America's first addiction treatment field was the founding of the American Association for the Cure of Inebriety in 1870 and its publication of the first issue of the Journal of Inebriety in 1876. One of the most contentious issues among the various branches of this new professional field was the question of the use of "reformed men" as physicians, managers and attendants within treatment institutions. This article describes the employment of recovering physicians within one 19th century addiction treatment franchise- the Keeley Institutes--and documents the nature of the professional debate stirred by what was then a controversial practice. PMID- 10809517 TI - Outcome and six month follow up of patients after Ultra Rapid Opiate Detoxification (UROD). AB - OBJECTIVES: (1) Evaluate the safety and efficacy of Ultra Rapid Opiate Detoxification (UROD); and (2) evaluate six month outcome data of patients choosing this method. DESIGN: Two center parallel group clinical trial. No grant funding. SETTING: Two academic medical centers. PARTICIPANTS: Ninety-three men and 27 women, aged 18 to 55 years, with opiate dependency self selected to undergo detoxification. INTERVENTIONS: UROD followed by naltrexone maintenance and an aftercare program. UROD and aftercare costs were the responsibility of the patients and/or their significant others. MAIN OUTCOMES MEASURE: (1) Completion of UROD as determined by a non reactive response to a naloxone challenge test under anesthesia and non reactive response to naltrexone administration before discharge. (2) Patient outcome as determined at six month follow up of UROD patients' self reported relapse free status confirmed by urine drug screen, significant other reports, and/or therapist reports. RESULTS: One hundred percent successful detoxification with UROD with low morbidity and no mortality. Relapse data were available for 111/123 procedures performed (90%), with 61/111 patients (55%) with reported relapse free status at the six month follow up interval. CONCLUSIONS: For individuals who are addicted to opioids, the Ultra Rapid Opiate Detoxification method appears to be a viable treatment option. PMID- 10809518 TI - Methadone dose and retention during treatment of heroin addicts with Axis I psychiatric comorbidity. AB - We studied 90 opioid-dependent subjects, 38 with one or more additional Axis I diagnosis and 52 with no psychiatric comorbidity. There were significant differences between these two groups regarding the methadone dose required for clinical stabilization, but not in the rate of retention in treatment. Dual Diagnosis patients, those with psychiatric comorbidity, required an average stabilization dose of 154 +/- 84 of methadone compared to 99 +/- 49 mg/day for patients whose only Axis I diagnosis was Opioid Dependence. In the 990-day period considered there were no differences between the two groups of patients in terms of retention in treatment. PMID- 10809519 TI - Identifying office resource needs of Canadian physicians to help prevent, assess and treat patients with substance use and pathological gambling disorders. AB - A study was conducted to determine whether there is a need for office-based resources to assist physicians in the prevention, assessment and/or management of patients, self and peers with substance use (excluding alcohol and tobacco) or pathological gambling disorders. The needs assessment was undertaken in three parts. Survey information was collected from 54 respondents including Executive Directors of the Canadian Medical Association's national affiliates and provincial/territorial Divisions, Deputy Ministers of Health, Registrars of provincial/territorial licensing bodies and Deans or Associate Deans of Continuing Medical Education programs in universities. Key informant interviews were conducted with 22 "experts" in the field of substance use and/or pathological gambling disorders. Focus groups were held in Vancouver, Toronto, Ottawa, Montreal and Halifax with 34 physicians who are interested in and whose caseload included patients with substance use and/or gambling disorders. Results suggest that current resources for both substance use and pathological gambling disorders are inadequate for physicians because there are gaps in the types activities and resources available, existing resources have not been effectively diffused or disseminated to physicians and training is needed to complement these resources to encourage proper implementation. PMID- 10809520 TI - Characteristics of physicians presenting for assessment at a behavioral health center. AB - Physician health and impairment have been of considerable interest in recent years. This study contributes detailed clinical data to the existing body of knowledge, by drawing from a sample of physicians assessed at a behavioral health center over a two year time frame. Demographic, referral, and clinical data were gathered using a systematic medical record review procedure, based on 108 physicians who were evaluated within an intensive multidisciplinary assessment program. The majority suffered from active substance use disorders (52.8%), with other psychiatric disorders (29.6%), and substance use disorders in remission (17.6%) the other largest categories. Of those with an active substance use disorder, primary drugs of choice were alcohol and prescription opiates. Over half had comorbid psychiatric disorders (Axis I, II, or both). Significant relationship, employment, and emotional problems were found in all three groups. The significant increase in presentation and/or detection of psychiatric and behavioral problems, both comorbid with and not substance use related, confirms the need for a revision and expansion of views about physicians' behavioral health concerns. PMID- 10809521 TI - The relationship between adolescent smoking and drinking and likelihood estimates of illicit drug use. AB - Using data from a recent national survey of adolescent substance use, the present work examined whether adolescents with different patterns of alcohol and cigarette use differed in their estimates of the likelihood they would use an illegal drug in the future. While nonusers of either substance were the most likely to indicate that they would never use drugs in the future, users of both substances were the most likely to indicate that they would use drugs. In addition, while users of both were most likely to indicate that they were likely to use illegal substances, only-smokers were more likely than only-drinkers to indicate that they were likely to use such substances in the future. Results are discussed in terms of the gateway theory of drug sequencing and cognitive precursors of experimentation with illegal substances. PMID- 10809522 TI - Utilization of services by persons discharged from involuntary chemical dependency treatment. AB - This report compares services utilization pre-admission and post-discharge in 735 consecutive persons involuntarily committed to a chemical dependency treatment program in Washington State. Patients entering treatment were in their late 30s, had multiple health problems, previous arrests for misdemeanors or felonies, and minimal structured daily activities. Post discharge, there were decreases in the use of costly acute care services including detox, psychiatric hospitalization, and mental health crisis services. Patients who completed the program were less likely to use acute care services and were more likely to participate in outpatient treatment after discharge. The overall death rate of 29.4 per 1000 persons per year was 4 times greater than the age adjusted death rate for the US adult population. Further studies of other involuntary chemical dependency treatment programs are needed to evaluate the results of this report. PMID- 10809523 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 10809524 TI - Implementing the ASAM Criteria in community treatment centers in Illinois: opportunities and challenges. American Society of Addiction Medicine. AB - The incorporation of the ASAM Criteria into addiction treatment centers procedure has afforded us an enormous opportunity to add credibility to our treatment. Herein lies perhaps the strongest argument for these criteria. The ASAM Criteria encourage addiction treatment centers to establish themselves as healthcare providers in their own right. The task is now for Illinois addiction treatment providers; particularly community based providers, to prove our credibility through the legitimate use of the criteria. Since most, if not all, of our patients are without means and without knowledge on quality treatment, they become easy targets for substandard or at least unimaginative and uninspired treatment. It is not that community based treatment centers are, by nature, unprincipled treatment warehouses. But it certainly can be said that our centers are frequently the last to change our treatment practices. Certainly we have become "the last resort" for the courts, overburdened mental health centers and overpopulated homeless shelters with little emphasis on clinical quality of care. Overcoming such programmed ways of viewing treatment was difficult specifically for Triangle Center and generally for community treatment providers throughout Illinois. In fact, I dare say that this transition has not occurred in total as many still passively resist this opportunity. Yet others have viewed this change as the breath of fresh air that our profession has long sought. Such variance in opinion ultimately proves that implementation of the ASAM Criteria, in a community based system, is dependent on the clinician and administrators willingness to understand, recognize and apply the Criteria. PMID- 10809525 TI - Practice guidelines: an ASAM-AMBHA joint statement. American Society of Addiction Medicine and the American Managed Behavioral Healthcare Association. PMID- 10809526 TI - Public policy statement on centralized credentialing systems and the physician in recovery. PMID- 10809529 TI - Molecular library technologies at the millennium. PMID- 10809528 TI - Public policy statement on reporting of patient information related to fitness for driving or other potentially dangerous activities. PMID- 10809530 TI - Bioinformatics in the pre- and post-genomic eras. PMID- 10809527 TI - Public policy statement on confidentiality in physician illness. PMID- 10809531 TI - Wilms' tumour and Fanconi anaemia: an unusual association. PMID- 10809532 TI - Ascites in a premature baby due to parenteral nutrition from an umbilical venous catheter. PMID- 10809533 TI - Prenatal care applications. PMID- 10809534 TI - Reduced interneuronal space in schizophernia. PMID- 10809535 TI - [Cerebrotendinous xanthomatosis in a 12-year-old patient with growth disorder]. PMID- 10809536 TI - [Leiomyoma of the inferior vena cava: diagnostic imaging and transfemoral bioptic confirmation]. PMID- 10809537 TI - [Intraosseus meningioma: a case report and differential diagnosis]. PMID- 10809538 TI - [Intragastric penetration as a local complication after performance of gastric banding]. PMID- 10809539 TI - Nucleotide insertions and deletions within the homopolymeric runs of adenines and thymidines of BCL10 cDNAs in normal peripheral blood leukocytes. PMID- 10809541 TI - Paper alert. Biotechnology. PMID- 10809540 TI - Treatment outcome in infant acute lymphoblastic leukemia. Children Leukemia Cooperative Group--EORTC. European Organization for Research and Treatment of Cancer. PMID- 10809542 TI - Web alert. Plant biotechnology. Biochemical engineering. PMID- 10809543 TI - Plant biotechnology. The ins and outs of a new green revolution. PMID- 10809544 TI - Biochemical engineering. Bridging the gap between gene and product. PMID- 10809545 TI - Analgesic effects of sweet solutions and pacifiers in term neonates. Surely evidence is not needed to justify cuddling babies in pain. PMID- 10809546 TI - Analgesic effects of sweet solutions and pacifiers in term neonates. Use of pacifier may modify responses measured on rating scale. PMID- 10809547 TI - Analgesic effects of sweet solutions and pacifiers in term neonates. Measures of chest pain must be validated in young children. PMID- 10809548 TI - Analgesic effects of sweet solutions and pacifiers in term neonates. Trial of drug treatment to alleviate pain in neonatal intubation is needed. PMID- 10809549 TI - Diagnose and be damned. Doctors must remember their rights and obligations to infants and children. PMID- 10809550 TI - Diagnose and be damned. Inquiry should be held into doctors involved in child abuse investigations. PMID- 10809551 TI - Diagnose and be damned. Management of CFS in children is not contentious. PMID- 10809552 TI - How much to do at the accident scene? Anaesthetists are best people to provide prehospital airway management. PMID- 10809553 TI - How much to do at the accident scene? Prehospital intervention prolong prehospital time. PMID- 10809554 TI - Predicted impact of intravenous thrombolysis. Another trial is needed. PMID- 10809555 TI - Using anticoagulation or aspirin to prevent stroke. Anticoagulation has a major role in primary prevention of stroke in general practice. PMID- 10809556 TI - Using anticoagulation or aspirin to prevent stroke. Study does not have the power to show that aspirin is as good as anticoagulation. PMID- 10809557 TI - Using anticoagulation or aspirin to prevent stroke. Results of the study cannot be generalised to the general practice population. PMID- 10809558 TI - Using anticoagulation or aspirin to prevent stroke. Aspirin is the logical choice for non-rheumatic atrial fibrillation. PMID- 10809559 TI - Using anticoagulation or aspirin to prevent stroke. Patients should be assessed for risk factors before deciding on prophylaxis. PMID- 10809560 TI - Pituitary surgery for acromegaly. PMID- 10809561 TI - New approaches to bacterial vaccine development. Proceedings of the 3rd International Meeting. Munich, Germany, 19-22 May 1999. PMID- 10809562 TI - 1st International Conference in Israel on Respiratory Rehabilitation in Patients with Chronic Respiratory Failure: from the ICU to Home. Safad, Israel, March 14 16, 2000. Abstracts. PMID- 10809563 TI - Proceedings of the European Seminars in Respiratory Medicine--Update 1999. St Moritz, Switzerland, July 2-6, 1999. Part 2. PMID- 10809564 TI - Noninvasive ventilation in COPD patients with chronic respiratory failure--con. PMID- 10809565 TI - Experience with the preperitoneal 'plug and patch' inguinal hernia repair. PMID- 10809566 TI - Consultant surgeons' opinion of the skills required of basic surgical trainees. PMID- 10809567 TI - Postoperative peritonitis originating from the duodenum: operative management by intubation and continuous intraluminal irrigation. PMID- 10809568 TI - A 14-year experience with 6 cm as a criterion for surgical treatment of abdominal aortic aneurysm. PMID- 10809569 TI - Retrosternal goitre. PMID- 10809570 TI - Management of liver trauma. PMID- 10809571 TI - Simultaneous aortic aneurysm repair and colonic surgery. PMID- 10809572 TI - A hernia sac cannot be found at operation. PMID- 10809573 TI - A hernia sac cannot be found at operation. PMID- 10809574 TI - A hernia sac cannot be found at operation. PMID- 10809575 TI - Expression and clinical significance of apolipoprotein D in male breast cancer and gynaecomastia. PMID- 10809576 TI - Randomized clinical trial of early laparoscopy in the management of acute non specific abdominal pain. PMID- 10809577 TI - Randomized clinical trial of early laparoscopy in the management of acute non specific abdominal pain. PMID- 10809578 TI - Randomized clinical trial of early laparoscopy in the management of acute non specific abdominal pain. PMID- 10809579 TI - ISMB '99. Proceedings of the 7th International Conference on Intelligent Systems for Molecular Biology. Heidelberg, Germany, August 6-10, 1999. PMID- 10809580 TI - Proceedings of the 3rd International Conference on TGF-beta: Biological Mechanisms and Clinical Applications. Bethesda, Maryland, USA. 20-22 September 1999. PMID- 10809581 TI - Does prophylactic octreotide benefit patients undergoing elective pancreatic resection? PMID- 10809582 TI - Simplified and rapid assay for detecting the 3'A mutation of the SDF-1 gene. PMID- 10809584 TI - Retroperitoneal laparoscopy. PMID- 10809583 TI - Triglyceride and Lp(a) concentrations in hyperapobetalipoproteinemia. PMID- 10809585 TI - Testicular prosthesis placement: new technique. PMID- 10809586 TI - National bias in citations in urology journals: parochialism or availability? PMID- 10809587 TI - Hazards of aspirin withdrawal before transurethral prostatectomy. PMID- 10809588 TI - Current literature in diabetes. PMID- 10809589 TI - Acute hydrocarbon pneumonia. PMID- 10809590 TI - Polymer supported reagents in synthesis: preparation of bicyclo[2.2.2] octane derivates via Tandem Michael addition reactions and subsequent combinatorial decoration. PMID- 10809591 TI - Perspective Articles on the utility and application of solution-phase combinatorial chemistry. PMID- 10809592 TI - An international conference with a high ambition. Cellular Responses to Oxidative and Osmotic Stress: Sensing, Signalling and Gene Expression, Egmond aan Zee, The Netherlands, 28 August-1 September 1999. PMID- 10809593 TI - GM crops: environmental risks and non-target effects. PMID- 10809594 TI - [100 years of organized cancer research--successes, hopes, disappointments]. PMID- 10809595 TI - Anaesthesiology directors: more support on the way as Association of Anesthesia Clinical Directors goes international. PMID- 10809596 TI - Histological assessment of vertebral bone in a Down's syndrome adult with osteoporosis. PMID- 10809597 TI - Kikuchi's disease in axillary lymph nodes draining breast carcinoma. PMID- 10809598 TI - Adenomatoid hyperplasia of the Bowman's capsule epithelium in association with primary pulmonary primitive neuroectodermal tumour (PNET) PMID- 10809599 TI - Malignant carcinoid tumour arising in a mature teratoma of the fallopian tube. PMID- 10809600 TI - Melanotic progonoma as a component of ovarian teratoma. PMID- 10809601 TI - Oncocytic lipoadenoma of the parotid gland. PMID- 10809602 TI - Primary acinic cell-like carcinoma of the breast--a variant with good prognosis? PMID- 10809603 TI - Peri-prosthetic femoral shaft fractures treated with plate and cable fixation. PMID- 10809604 TI - Further decline of hepatitis B surface antigen (HBsAg) prevalence in Japan. PMID- 10809605 TI - Advances in our understanding of the treatment of painful bone metastasis. PMID- 10809606 TI - Evidence of increased expression of synaptic basal lamina protein agrin in glomerulosclerosis. PMID- 10809607 TI - Transient decrease of serum bicarbonate levels with Sevelamer hydrochloride as the phosphate binder. PMID- 10809608 TI - [Detection of metastatic microfoci on the omental milk spots as the initial implantation sites for malignant gastric cancer cells in peritoneal dissemination: a method by using RT-PCR]. PMID- 10809609 TI - [Evaluation of p21 and lung resistance protein (LRP) as a factor suggesting breast cancer recurrence]. PMID- 10809610 TI - Coronary-artery stenting in acute myocardial infarction. PMID- 10809611 TI - Coronary-artery stenting in acute myocardial infarction. PMID- 10809612 TI - Coronary-artery stenting in acute myocardial infarction. PMID- 10809613 TI - Improved clinical outcome after widespread use of coronary-artery stenting in Canada. PMID- 10809614 TI - Idiopathic chronic pericardial effusion. PMID- 10809615 TI - Idiopathic chronic pericardial effusion. PMID- 10809616 TI - Transient enhanced uptake of 123I-metaiodobenzylguanidine in the contralateral adrenal region after resection of an adrenal pheochromocytoma. PMID- 10809617 TI - Domestic violence. PMID- 10809618 TI - Domestic violence. PMID- 10809619 TI - The scope of practice in primary care. PMID- 10809620 TI - The scope of practice in primary care. PMID- 10809621 TI - The scope of practice in primary care. PMID- 10809622 TI - Making the diagnosis of subarachnoid hemorrhage. PMID- 10809623 TI - Making the diagnosis of subarachnoid hemorrhage. PMID- 10809624 TI - German parliament agrees on limits to bioethics inquiry. PMID- 10809625 TI - Ontario joins the genomics goldrush. PMID- 10809626 TI - Director of Wellcome centre resigns over damning report. PMID- 10809627 TI - UK ethicists back use of stem cells. PMID- 10809628 TI - Human parvovirus B19 and renal disease? PMID- 10809629 TI - Glucose, free fatty acids, and insulin secretion. PMID- 10809630 TI - Sweetening agents. PMID- 10809631 TI - A physiological function for thyrocalcitonin. PMID- 10809632 TI - [The organization of the female reproductive system in the parasitic mite Myobia murismusculi (Trombidiformes: Myobiidae)]. AB - The general morphology of the internal genital organs in Myobia murismusculi females have been studied by means of light and electron microscopy. It is shown, that their reproductive system is composed of the single ovary, long complex oviduct, cuticular vagina and receptaculum seminis. The apical region of the ovary is formed by the numerous processes of gonadian somatic cells. These cells generate the ovarian sheath and probably take place in oocyte's metabolism. Any nutritive cells in the Myobia ovary are absent. The vitellogenetic oocytes develop in the ovarian pouches like in other trombidiform mites. The oviduct consists of three parts, each of them has a specific ultrastructure and type of secretory product. Two proximal parts of the oviduct produce the egg-shell components, while the third distal part never contains oocytes. It could be suggested that this part produces a special secret onto the oocyte surface to glue the oocytes to the hairs of host. PMID- 10809634 TI - The use of chest drains. PMID- 10809633 TI - [Brachylepis gen. n.--a new genus of cestodes (Cyclophyllidea: Hymenolepididae) from shrews in Siberia and the Far East]. AB - A new genus Brachylepis gen. n. hymenolepididean cestodes associated with common shrews (Sorex) in Siberia and Russian Far East is created. These genus includes three species removed from the genus Mathevolepis: Brachylepis morosovi (Karpenko, 1994) comb. n. (type species), B. sorextscherskii (Morosov, 1957) comb. n. and B. triovaria Karpenko, 1990) comb. n. The new genus is characterized by the gradual strobila with a little number of segments, unarmed scolex with apical glandular organ, by lack of internal seminal vesicle and differential seminal receptacle, by form of uterus as horse shoe in young worms and buble-like in mature worms. A tendency of inverse proportional dependence between the number of uterus segments in the strobila and the number of hexacanths in the uterus has been noticed. The table of differential characters for the species of the genus Brachylepis is proposed. PMID- 10809635 TI - Environmental health: responding to the call... PMID- 10809636 TI - [Tuberculosis in HIV infection]. PMID- 10809637 TI - Comment on: single fraction radiotherapy for bone metastases. Are all questions answered? PMID- 10809638 TI - [The Gr. T. Popa University of Medicine and Pharmacy in Iasi on the brink of the year 2000. On the 120th anniversary of its existence (I)]. PMID- 10809639 TI - New neurons: extraordinary evidence or extraordinary conclusion? PMID- 10809640 TI - Ecology. Does biodiversity help fend off invaders? PMID- 10809641 TI - Cancer research. Caution raised about possible new drug. PMID- 10809642 TI - Advanced technology program. Grants evoke squeals of delight and anger. PMID- 10809643 TI - Evolution. Stretching the reign of early animals. PMID- 10809644 TI - Biomedical policy. New minister hopes to create an Italian NIH. PMID- 10809645 TI - Antibiotic resistance. Superbugs on the hoof? PMID- 10809646 TI - Genomics. Money and machines fuel China's push in sequencing. PMID- 10809647 TI - American Association of Physical Anthropologists. From field to lab, new insights on being human. PMID- 10809648 TI - Genomes 2000. Intimate portraits of bacterial nemeses. PMID- 10809649 TI - Education for a biocomplex future. PMID- 10809650 TI - Essays on science and society. Not (just) in Kansas anymore. PMID- 10809651 TI - Social science. Shrewd investments. PMID- 10809652 TI - Plant biology. Lighting up the nucleus. PMID- 10809653 TI - Premotor commands encode monocular eye movements. Has Hering been hooked? PMID- 10809654 TI - Discontinuation of anticytomegalovirus therapy in patients with HIV infection and cytomegalovirus retinitis. PMID- 10809655 TI - "Filling-in" and afterimages. PMID- 10809656 TI - [Polish Symposium on Paediatric Oncology. Proceedings, June 17-19, 1998]. PMID- 10809657 TI - Referring to the posterior fossa craniectomy and tonsillar resection in order to treat Chiari I malformation with syringomyelia. PMID- 10809658 TI - Transient mutism after brain stem infarction. PMID- 10809659 TI - Antihypertensive therapy for nondiabetic nephropathy. PMID- 10809660 TI - Homage to Takis S. Papas, 1935-1999. PMID- 10809661 TI - More on management of choroid plexus cysts in the mid-trimester fetus--Reply. PMID- 10809662 TI - Sir2 links chromatin silencing, metabolism, and aging. PMID- 10809663 TI - Protein-protein interactions define specificity in signal transduction. PMID- 10809664 TI - A core SMRT corepressor complex containing HDAC3 and TBL1, a WD40-repeat protein linked to deafness. AB - The corepressor SMRT mediates repression by thyroid hormone receptor (TR) as well as other nuclear hormone receptors and transcription factors. Here we report the isolation of a novel SMRT-containing complex from HeLa cells. This complex contains transducin beta-like protein 1 (TBL1), whose gene is mutated in human sensorineural deafness. It also contains HDAC3, a histone deacetylase not previously thought to interact with SMRT. TBL1 displays structural and functional similarities to Tup1 and Groucho corepressors, sharing their ability to interact with histone H3. In vivo, TBL1 is bridged to HDAC3 through SMRT and can potentiate repression by TR. Intriguingly, loss-of-function TRbeta mutations cause deafness in mice and humans. These results define a new TR corepressor complex with a physical link to histone structure and a potential biological link to deafness. PMID- 10809665 TI - The Drosophila brahma complex is an essential coactivator for the trithorax group protein zeste. AB - The trithorax group (trxG) of activators and Polycomb group (PcG) of repressors are believed to control the expression of several key developmental regulators by changing the structure of chromatin. Here, we have sought to dissect the requirements for transcriptional activation by the Drosophila trxG protein Zeste, a DNA-binding activator of homeotic genes. Reconstituted transcription reactions established that the Brahma (BRM) chromatin-remodeling complex is essential for Zeste-directed activation on nucleosomal templates. Because it is not required for Zeste to bind to chromatin, the BRM complex appears to act after promoter binding by the activator. Purification of the Drosophila BRM complex revealed a number of novel subunits. We found that Zeste tethers the BRM complex via direct binding to specific subunits, including trxG proteins Moira (MOR) and OSA. The leucine zipper of Zeste mediates binding to MOR. Interestingly, although the Imitation Switch (ISWI) remodelers are potent nucleosome spacing factors, they are dispensable for transcriptional activation by Zeste. Thus, there is a distinction between general chromatin restructuring and transcriptional coactivation by remodelers. These results establish that different chromatin remodeling factors display distinct functional properties and provide novel insights into the mechanism of their targeting. PMID- 10809666 TI - MEI-1/MEI-2 katanin-like microtubule severing activity is required for Caenorhabditis elegans meiosis. AB - The Caenorhabditis elegans meiotic spindle is morphologically distinct from the first mitotic spindle, yet both structures form in the same cytoplasm approximately 20 minutes apart. The mei-1 and mei-2 genes of C. elegans are required for the establishment of the oocyte meiotic spindle but are not required for mitotic spindle function. mei-1 encodes an AAA ATPase family member with similarity to the p60 catalytic subunit of the heterodimeric sea urchin microtubule-severing protein, katanin. We report that mei-2 encodes a 280-amino acid protein containing a region similar to the p80-targeting subunit of katanin. MEI-1 and MEI-2 antibodies decorate the polar ends of meiotic spindle microtubules and meiotic chromatin. We find that the subcellular location of MEI 2 depends on wild-type mei-1 activity and vice versa. These experiments, combined with MEI-1 and MEI-2's similarity to p60 and p80 katanin, suggest that the C. elegans proteins function as a complex. In support of this idea, MEI-1 and MEI-2 physically associate in HeLa cells. Furthermore, co-expression of MEI-1 and MEI-2 in HeLa cells results in the disassembly of microtubules. These data lead us to conclude that MEI-1/MEI-2 microtubule-severing activity is required for meiotic spindle organization in C. elegans. PMID- 10809667 TI - MutS homolog 4 localization to meiotic chromosomes is required for chromosome pairing during meiosis in male and female mice. AB - Msh4 (MutS homolog 4) is a member of the mammalian mismatch repair gene family whose members are involved in postreplicative DNA mismatch repair as well as in the control of meiotic recombination. In this report we show that MSH4 has an essential role in the control of male and female meiosis. We demonstrate that MSH4 is present in the nuclei of spermatocytes early in prophase I and that it forms discrete foci along meiotic chromosomes during the zygotene and pachytene stages of meiosis. Disruption of the Msh4 gene in mice results in male and female sterility due to meiotic failure. Although meiosis is initiated in Msh4 mutant male and female mice, as indicated by the chromosomal localization of RAD51 and COR1 during leptonema/zygonema, the chromosomes fail to undergo normal pairing. Our results show that MSH4 localization on chromosomes during the early stages of meiosis is essential for normal chromosome synapsis in prophase I and that it acts in the same pathway as MSH5. PMID- 10809668 TI - Pre-mRNA splicing alters mRNP composition: evidence for stable association of proteins at exon-exon junctions. AB - We provide direct evidence that pre-mRNA splicing alters mRNP protein composition. Using a novel in vitro cross-linking approach, we detected several proteins that associate with mRNA exon-exon junctions only as a consequence of splicing. Immunoprecipitation experiments suggested that these proteins are part of a tight complex around the junction. Two were identified as SRm160, a nuclear matrix-associated splicing coactivator, and hPrp8p, a core component of U5 snRNP and spliceosomes. Glycerol gradient fractionation showed that a subset of these proteins remain associated with mRNA after its release from the spliceosome. These results demonstrate that the spliceosome can leave behind signature proteins at exon-exon junctions. Such proteins could influence downstream metabolic events in vivo such as mRNA transport, translation, and nonsense mediated decay. PMID- 10809669 TI - Hfq (HF1) stimulates ompA mRNA decay by interfering with ribosome binding. AB - The adaptation of mRNA stability to environmental changes is a means of cells to adjust the level of gene expression. The Escherichia coli ompA mRNA has served as one of the paradigms for regulated mRNA decay in prokaryotes. The stability of the transcript is known to be correlated inversely with the bacterial growth rate. Thus, the regulation of ompA mRNA stability meets the physiological needs to adjust the level of ompA expression to the rate of cell division. Recently, host factor I (Hfq/HF1) was shown to be involved in the regulation of ompA mRNA stability under slow growth conditions. Here, we present the first direct demonstration that 30S ribosomes bound to the ompA 5'-UTR protect the transcript from RNase E cleavage in vitro. However, the 30S protection was found to be abrogated in the presence of Hfq. Toeprinting and in vitro translation assays revealed that translation of ompA is repressed in the presence of Hfq. These in vitro studies are corroborated by in vivo expression studies demonstrating that the reduced synthesis rate of OmpA effected by Hfq results in functional inactivation of the ompA mRNA. The data are discussed in terms of a model wherein Hfq regulates the stability of ompA mRNA by competing with 30S ribosomes for binding to the ompA 5'-UTR. PMID- 10809670 TI - A subtilisin-like serine protease involved in the regulation of stomatal density and distribution in Arabidopsis thaliana. AB - Stomata are specialized cellular structures in the epidermis of aerial plant organs that control gas exchange (H(2)O release and CO(2) uptake) between leaves and the atmosphere by modulating the aperture of a pore flanked by two guard cells. Stomata are nonrandomly distributed, and their density is controlled by endogenous and environmental factors. To gain insight into the molecular mechanisms regulating stomatal distribution, Arabidopsis thaliana mutants with altered stomatal characteristics were isolated and examined. The sdd1-1 mutant exhibits a two- to fourfold increase of stomatal density and formation of clustered stomata (i.e., stomata that are not separated by intervening pavement cells), whereas the internal leaf architecture is not altered. The SDD1 gene was identified by map-based cloning. It encodes a subtilisin-like serine protease related to prokaryotic and eukaryotic proteins. We propose that SDD1 acts as a processing protease involved in the mediation of a signal that controls the development of cell lineages that lead to guard cell formation. PMID- 10809671 TI - The ShcA phosphotyrosine docking protein sensitizes cardiovascular signaling in the mouse embryo. AB - The ShcA gene products have served as a model for the analysis of phosphotyrosine recognition domains, and for the functions of docking proteins during tyrosine kinase signaling. Here we show that ShcA is primarily expressed in the cardiovascular system during early mouse embryogenesis and regulates both heart development and establishment of mature blood vessels. Targeted mutation suggests that the ShcA adaptor is a pivotal target of tyrosine kinases that selectively potentiates activation of the MAP kinase pathway in the remodeling vasculature. Biochemical analysis of mutant cells shows that ShcA sensitizes cells to growth factor-induced MAP kinase activation, and also organizes cytoskeletal rearrangement in response to the extracellular matrix. ShcA may therefore orchestrate complex interactions within the vascular compartment by rendering cells permissive to respond to soluble and adhesive external cues. PMID- 10809672 TI - SPC4/PACE4 regulates a TGFbeta signaling network during axis formation. AB - In vertebrates, specification of anteroposterior (A/P) and left-right (L/R) axes depends on TGFbeta-related signals, including Nodal, Lefty, and BMPs. Endoproteolytic maturation of these proteins is probably mediated by the proprotein convertase SPC1/Furin. In addition, precursor processing may be regulated by related activities such as SPC4 (also known as PACE4). Here, we show that a proportion of embryos lacking SPC4 develop situs ambiguus combined with left pulmonary isomerism or complex craniofacial malformations including cyclopia, or both. Gene expression analysis during early somite stages indicates that spc4 is genetically upstream of nodal, pitx2, lefty1, and lefty2 and perhaps maintains the balance between Nodal and BMP signaling in the lateral plate that is critical for L/R axis formation. Furthermore, genetic interactions between nodal and spc4, together with our analysis of chimeric embryos, strongly suggest that during A/P axis formation, SPC4 acts primarily in the foregut. These findings establish an important role for SPC4 in patterning the early mouse embryo. PMID- 10809673 TI - CIITA is a transcriptional coactivator that is recruited to MHC class II promoters by multiple synergistic interactions with an enhanceosome complex. AB - By virtue of its control over major histocompatibility complex class II (MHC-II) gene expression, CIITA represents a key molecule in the regulation of adaptive immune responses. It was first identified as a factor that is defective in MHC-II deficiency, a hereditary disease characterized by the absence of MHC-II expression. CIITA is a highly regulated transactivator that governs all spatial, temporal, and quantitative aspects of MHC-II expression. It has been proposed to act as a non-DNA-binding transcriptional coactivator, but evidence that it actually functions at the level of MHC-II promoters was lacking. By means of chromatin immunoprecipitation assays, we show here for the first time that CIITA is physically associated with MHC-II, as well as HLA-DM, Ii, MHC-I, and beta(2)m promoters in vivo. To dissect the mechanism by which CIITA is recruited to the promoter, we have developed a DNA-dependent coimmunoprecipitation assay and a pull-down assay using immobilized promoter templates. We demonstrate that CIITA recruitment depends on multiple, synergistic protein-protein interactions with DNA-bound factors constituting the MHC-II enhanceosome. CIITA therefore represents a paradigm for a novel type of regulatory and gene-specific transcriptional cofactor. PMID- 10809675 TI - Cloning of a mucin-desulfating sulfatase gene from Prevotella strain RS2 and its expression using a Bacteroides recombinant system. AB - A gene encoding the mucin-desulfating sulfatase in Prevotella strain RS2 has been cloned, sequenced, and expressed in an active form. A 600-bp PCR product generated using primers designed from amino acid sequence data was used to isolate a 5,058-bp genomic DNA fragment containing the mucin-desulfating sulfatase gene. A 1,551-bp open reading frame encoding the sulfatase proprotein was identified, and the deduced 517-amino-acid protein minus its signal sequence corresponded well with the published mass of 58 kDa estimated by denaturing gel electrophoresis. The sulfatase sequence showed homology to aryl- and nonarylsulfatases with different substrate specificities from the sulfatases of other organisms. No sulfatase activity could be detected when the sulfatase gene was cloned into Escherichia coli expression vectors. However, cloning the gene into a Bacteroides expression vector did produce active sulfatase. This is the first mucin-desulfating sulfatase to be sequenced and expressed. A second open reading frame (1,257 bp) was identified immediately upstream from the sulfatase gene, coding in the opposite direction. Its sequence has close homology to iron sulfur proteins that posttranslationally modify other sulfatases. By analogy, this protein is predicted to catalyze the modification of a serine group to a formylglycine group at the active center of the mucin-desulfating sulfatase, which is necessary for enzymatic activity. PMID- 10809674 TI - A biochemical mechanism for nonrandom mutations and evolution. PMID- 10809676 TI - Identification of an effector specificity subregion within the aromatic responsive regulators DmpR and XylR by DNA shuffling. AB - The Pseudomonas derived sigma(54)-dependent regulators DmpR and XylR control the expression of genes involved in catabolism of aromatic compounds. Binding to distinct, nonoverlapping groups of aromatic effectors controls the activities of these transcriptional activators. Previous work has derived a common mechanistic model for these two regulators in which effector binding by the N-terminal 210 residues (the A-domain) of the protein relieves repression of an intrinsic ATPase activity essential for its transcription-promoting property and allows productive interaction with the transcriptional apparatus. Here we dissect the A-domains of DmpR and XylR by DNA shuffling to identify the region(s) that mediates the differences in the effector specificity profiles. Analysis of in vivo transcription in response to multiple aromatic effectors and the in vitro phenol binding abilities of regulator derivatives with hybrid DmpR/XylR A-domains reveals that residues 110 to 186 are key determinants that distinguish the effector profiles of DmpR and XylR. Moreover, the properties of some mosaic DmpR/XylR derivatives reveal that high-affinity aromatic effector binding can be completely uncoupled from the ability to promote transcription. Hence, novel aromatic binding properties will only be translated into functional transcriptional activation if effector binding also triggers release of interdomain repression. PMID- 10809677 TI - Color-sensitive motility and methanol release responses in Rhodobacter sphaeroides. AB - Blue-light-induced repellent and demethylation responses, characteristic of behavioral adaptation, were observed in Rhodobacter sphaeroides. They were analyzed by computer-assisted motion analysis and through the release of volatile tritiated compounds from [methyl-(3)H]methionine-labeled cells, respectively. Increases in the stop frequency and the rate of methanol release were induced by exposure of cells to repellent light signals, such as an increase in blue- and a decrease in infrared-light intensity. At a lambda of >500 nm the amplitude of the methanol release response followed the absorbance spectrum of the photosynthetic pigments, suggesting that they function as photosensors for this response. In contrast to the previously reported motility response to a decrease in infrared light, the blue-light response reported here does not depend on the number of photosynthetic pigments per cell, suggesting that it is mediated by a separate sensor. Therefore, color discrimination in taxis responses in R. sphaeroides involves two photosensing systems: the photosynthetic pigments and an additional photosensor, responding to blue light. The signal generated by the former system could result in the migration of cells to a light climate beneficial for photosynthesis, while the blue-light system could allow cells to avoid too-high intensities of (harmful) blue light. PMID- 10809678 TI - Deletion analysis of the flagellar switch protein FliG of Salmonella. AB - The flagellar motor/switch complex, consisting of the three proteins FliG, FliM, and FliN, plays a central role in bacterial motility and chemotaxis. We have analyzed FliG, using 10-amino-acid deletions throughout the protein and testing the deletion clones for their motility and dominance properties and for interaction of the deletion proteins with the MS ring protein FliF. Only the N terminal 46 amino acids of FliG (segments 1 to 4) were important for binding to FliF; consistent with this, an N-terminal fragment consisting of residues 1 to 108 bound FliF strongly, whereas a C-terminal fragment consisting of residues 109 to 331 did not bind FliF at all. Deletions in the region from residues 37 to 96 (segments 4 to 9), 297 to 306 (segment 30), and 317 to 326 (segment 32) permitted swarming, though not at wild-type levels; all other deletions caused paralyzed or, more commonly, nonflagellate phenotype. Except for those near the N terminus, deletions had a dominant negative effect on wild-type cells. PMID- 10809679 TI - Interaction between FliE and FlgB, a proximal rod component of the flagellar basal body of Salmonella. AB - FliE is a flagellar basal body protein of Salmonella whose detailed location and function have not been established. A mutant allele of fliE, which caused extremely poor flagellation and swarming, generated extragenic suppressors, all of which mapped to flgB, one of four genes encoding the basal body rod; the fliE flgB pseudorevertants were better flagellated and swarmed better than the fliE parent, especially when the temperature was reduced from 37 to 30 degrees C. Motility of the pseudorevertants in liquid culture was markedly better than motility on swarm plates; we interpret this to mean that reduced flagellation is less deleterious at low viscous loads. Overproduction of the mutant FliE protein improved the motility of the parental fliE mutant and its pseudorevertants, though not to wild-type levels. Overproduction of suppressor FlgB (but not wild type FlgB) in the fliE mutant also resulted in improved motility. The second-site FlgB mutation by itself had no phenotype; cells swarmed as well as wild-type cells. When overproduced, wild-type FliE was dominant over FliE-V99G, but the reverse was not true; that is, overproduced FliE-V99G was not negatively dominant over wild-type FliE. We conclude that the mutant protein has reduced probability of assembly but, if assembled, functions relatively well. Export of the flagellar protein FlgD, which is known to be FliE dependent, was severely impaired by the FliE-V99G mutation but was significantly improved in the suppressor strains. The FliE mutation, V99G, was close to the C terminus of the 104-amino-acid sequence; the suppressing mutations in FlgB were all either G119E or G129D, close to the C terminus of its 138-amino-acid sequence. Affinity blotting experiments between FliE as probe and various basal body proteins as targets and vice versa revealed strong interactions between FliE and FlgB; much weaker interactions between FliE and other rod proteins were observed and probably derive from the known similarities among these proteins. We suggest that FliE subunits constitute a junction zone between the MS ring and the rod and also that the proximal rod structure consists of FlgB subunits. PMID- 10809680 TI - mRNA composition and control of bacterial gene expression. AB - The expression of any given bacterial protein is predicted to depend on (i) the transcriptional regulation of the promoter and the translational regulation of its mRNA and (ii) the synthesis and translation of total (bulk) mRNA. This is because total mRNA acts as a competitor to the specific mRNA for the binding of initiation-ready free ribosomes. To characterize the effects of mRNA competition on gene expression, the specific activity of beta-galactosidase expressed from three different promoter-lacZ fusions (P(spc)-lacZ, P(RNAI)-lacZ, and P(RNAII) lacZ) was measured (i) in a relA(+) background during exponential growth at different rates and (ii) in relA(+) and DeltarelA derivatives of Escherichia coli B/r after induction of a mild stringent or a relaxed response to raise or lower, respectively, the level of ppGpp. Expression from all three promoters was stimulated during slow exponential growth or at elevated levels of ppGpp and was reduced during fast exponential growth or at lower levels of ppGpp. From these observations and from other considerations, we propose (i) that the concentration of free, initiation-ready ribosomes is approximately constant and independent of the growth rate and (ii) that bulk mRNA made during slow growth and at elevated levels of ppGpp is less efficiently translated than bulk mRNA made during fast growth and at reduced levels of ppGpp. These features lead to an indirect enhancement in the expression of LacZ (or of any other protein) during growth in media of poor nutritional quality and at increased levels of ppGpp. PMID- 10809681 TI - Expression and characterization of the chitin-binding domain of chitinase A1 from Bacillus circulans WL-12. AB - Chitinase A1 from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III-like domains, and a C-terminal chitin-binding domain (ChBD). In order to study the biochemical properties and structure of the ChBD, ChBD(ChiA1) was produced in Escherichia coli using a pET expression system and purified by chitin affinity column chromatography. Purified ChBD(ChiA1) specifically bound to various forms of insoluble chitin but not to other polysaccharides, including chitosan, cellulose, and starch. Interaction of soluble chitinous substrates with ChBD(ChiA1) was not detected by means of nuclear magnetic resonance and isothermal titration calorimetry. In addition, the presence of soluble substrates did not interfere with the binding of ChBD(ChiA1) to regenerated chitin. These observations suggest that ChBD(ChiA1) recognizes a structure which is present in insoluble or crystalline chitin but not in chito oligosaccharides or in soluble derivatives of chitin. ChBD(ChiA1) exhibited binding activity over a wide range of pHs, and the binding activity was enhanced at pHs near its pI and by the presence of NaCl, suggesting that the binding of ChBD(ChiA1) is mediated mainly by hydrophobic interactions. Hydrolysis of beta chitin microcrystals by intact chitinase A1 and by a deletion derivative lacking the ChBD suggested that the ChBD is not absolutely required for hydrolysis of beta-chitin microcrystals but greatly enhances the efficiency of degradation. PMID- 10809682 TI - Environmental regulation of Bacillus subtilis sigma(D)-dependent gene expression. AB - The sigma(D) regulon of Bacillus subtilis is composed of genes encoding proteins for flagellar synthesis, motility, and chemotaxis. Concurrent analyses of sigma(D) protein levels and flagellin mRNA demonstrate that sigD expression and sigma(D) activity are tightly coupled during growth in both complex and minimal media, although they exhibit different patterns of expression. We therefore used the sigma(D)-dependent flagellin gene (hag) as a model gene to study the effects of different nutritional environments on sigma(D)-dependent gene expression. In complex medium, the level of expression of a hag-lacZ fusion increased exponentially during the exponential growth phase and peaked early in the transition state. In contrast, the level of expression of this reporter remained constant and high throughout growth in minimal medium. These results suggest the existence of a nutritional signal(s) that affects sigD expression and/or sigma(D) activity. This signal(s) allows for nutritional repression early in growth and, based on reconstitution studies, resides in the complex components of sporulation medium, as well as in a mixture of mono-amino acids. However, the addition of Casamino Acids to minimal medium results in a dose-dependent decrease in hag-lacZ expression throughout growth and the postexponential growth phase. In work by others, CodY has been implicated in the nutritional repression of several genes. Analysis of a codY mutant bearing a hag-lacZ reporter revealed that flagellin expression is released from nutritional repression in this strain, whereas mutations in the transition state preventor genes abrB, hpr, and sinR failed to elicit a similar effect during growth in complex medium. Therefore, the CodY protein appears to be the physiologically relevant regulator of hag nutritional repression in B. subtilis. PMID- 10809683 TI - Morphogenesis, adhesive properties, and antifungal resistance depend on the Pmt6 protein mannosyltransferase in the fungal pathogen candida albicans. AB - Protein mannosyltransferases (Pmt proteins) initiate O glycosylation of secreted proteins in fungi. We have characterized PMT6, which encodes the second Pmt protein of the fungal pathogen Candida albicans. The residues of Pmt6p are 21 and 42% identical to those of C. albicans Pmt1p and S. cerevisiae Pmt6p, respectively. Mutants lacking one or two PMT6 alleles grow normally and contain normal Pmt enzymatic activities in cell extracts but show phenotypes including a partial block of hyphal formation (dimorphism) and a supersensitivity to hygromycin B. The morphogenetic defect can be suppressed by overproduction of known components of signaling pathways, including Cek1p, Cph1p, Tpk2p, and Efg1p, suggesting a specific Pmt6p target protein upstream of these components. Mutants lacking both PMT1 and PMT6 are viable and show pmt1 mutant phenotypes and an additional sensitivity to the iron chelator ethylenediamine-di(o hydroxyphenylacetic acid). The lack of Pmt6p significantly reduces adherence to endothelial cells and overall virulence in a mouse model of systemic infection. The results suggest that Pmt6p regulates a more narrow subclass of proteins in C. albicans than Pmt1p, including secreted proteins responsible for morphogenesis and antifungal sensitivities. PMID- 10809684 TI - Fermentative metabolism of Bacillus subtilis: physiology and regulation of gene expression. AB - Bacillus subtilis grows in the absence of oxygen using nitrate ammonification and various fermentation processes. Lactate, acetate, and 2,3-butanediol were identified in the growth medium as the major anaerobic fermentation products by using high-performance liquid chromatography. Lactate formation was found to be dependent on the lctEP locus, encoding lactate dehydrogenase and a putative lactate permease. Mutation of lctE results in drastically reduced anaerobic growth independent of the presence of alternative electron acceptors, indicating the importance of NADH reoxidation by lactate dehydrogenase for the overall anaerobic energy metabolism. Anaerobic formation of 2,3-butanediol via acetoin involves acetolactate synthase and decarboxylase encoded by the alsSD operon. Mutation of alsSD has no significant effect on anaerobic growth. Anaerobic acetate synthesis from acetyl coenzyme A requires phosphotransacetylase encoded by pta. Similar to the case for lctEP, mutation of pta significantly reduces anaerobic fermentative and respiratory growth. The expression of both lctEP and alsSD is strongly induced under anaerobic conditions. Anaerobic lctEP and alsSD induction was found to be partially dependent on the gene encoding the redox regulator Fnr. The observed fnr dependence might be the result of Fnr-induced arfM (ywiD) transcription and subsequent lctEP and alsSD activation by the regulator ArfM (YwiD). The two-component regulatory system encoded by resDE is also involved in anaerobic lctEP induction. No direct resDE influence on the redox regulation of alsSD was observed. The alternative electron acceptor nitrate represses anaerobic lctEP and alsSD transcription. Nitrate repression requires resDE- and fnr-dependent expression of narGHJI, encoding respiratory nitrate reductase. The gene alsR, encoding a regulator potentially responding to changes of the intracellular pH and to acetate, is essential for anaerobic lctEP and alsSD expression. In agreement with its known aerobic function, no obvious oxygen or nitrate-dependent pta regulation was observed. A model for the regulation of the anaerobic fermentation genes in B. subtilis is proposed. PMID- 10809686 TI - Quantitative determination of metabolic fluxes during coutilization of two carbon sources: comparative analyses with Corynebacterium glutamicum during growth on acetate and/or glucose. AB - Growth of Corynebacterium glutamicum on mixtures of the carbon sources glucose and acetate is shown to be distinct from growth on either substrate alone. The organism showed nondiauxic growth on media containing acetate-glucose mixtures and simultaneously metabolized these substrates. Compared to those for growth on acetate or glucose alone, the consumption rates of the individual substrates were reduced during acetate-glucose cometabolism, resulting in similar total carbon consumption rates for the three conditions. By (13)C-labeling experiments with subsequent nuclear magnetic resonance analyses in combination with metabolite balancing, the in vivo activities for pathways or single enzymes in the central metabolism of C. glutamicum were quantified for growth on acetate, on glucose, and on both carbon sources. The activity of the citric acid cycle was high on acetate, intermediate on acetate plus glucose, and low on glucose, corresponding to in vivo activities of citrate synthase of 413, 219, and 111 nmol. (mg of protein)(-1). min(-1), respectively. The citric acid cycle was replenished by carboxylation of phosphoenolpyruvate (PEP) and/or pyruvate (30 nmol. [mg of protein](-1). min(-1)) during growth on glucose. Although levels of PEP carboxylase and pyruvate carboxylase during growth on acetate were similar to those for growth on glucose, anaplerosis occurred solely by the glyoxylate cycle (99 nmol. [mg of protein](-1). min(-1)). Surprisingly, the anaplerotic function was fulfilled completely by the glyoxylate cycle (50 nmol. [mg of protein](-1). min(-1)) on glucose plus acetate also. Consistent with the predictions deduced from the metabolic flux analyses, a glyoxylate cycle-deficient mutant of C. glutamicum, constructed by targeted deletion of the isocitrate lyase and malate synthase genes, exhibited impaired growth on acetate-glucose mixtures. PMID- 10809685 TI - Interacting regulatory circuits involved in orderly control of photosynthesis gene expression in Rhodobacter sphaeroides 2.4.1. AB - FnrL, the homolog of the global anaerobic regulator Fnr, is required for the induction of the photosynthetic apparatus in Rhodobacter sphaeroides 2.4.1. Thus, the precise role of FnrL in photosynthesis (PS) gene expression and its interaction(s) with other regulators of PS gene expression are of considerable importance to our understanding of the regulatory circuitry governing spectral complex formation. Using a CcoP and FnrL double mutant strain, we obtained results which suggested that FnrL is not involved in the transduction of the inhibitory signal, by which PS gene expression is "silenced," emanating from the cbb(3) oxidase encoded by the ccoNOQP operon under aerobic conditions. The dominant effect of the ccoP mutation in the FnrL mutant strain with respect to spectral complex formation under aerobic conditions and restoration of a PS positive phenotype suggested that inactivation of the cbb(3) oxidase to some extent bypasses the requirement for FnrL in the formation of spectral complexes. Additional analyses revealed that anaerobic induction of the bchE, hemN, and hemZ genes, which are involved in the tetrapyrrole biosynthetic pathways, requires FnrL. Thus, FnrL appears to be involved at multiple loci involved in the regulation of PS gene expression. Additionally, bchE was also shown to be regulated by the PrrBA two-component system, in conjunction with hemN and hemZ. These and other results to be discussed permit us to more accurately describe the role of FnrL as well as the interactions between the FnrL, PrrBA, and other regulatory circuits in the regulation of PS gene expression. PMID- 10809687 TI - A membrane-bound flavocytochrome c-sulfide dehydrogenase from the purple phototrophic sulfur bacterium Ectothiorhodospira vacuolata. AB - The amino acid sequence of Ectothiorhodospira vacuolata cytochrome c-552, isolated from membranes with n-butanol, shows that it is a protein of 77 amino acid residues with a molecular mass of 9,041 Da. It is closely related to the cytochrome subunit of Chlorobium limicola f. sp. thiosulfatophilum flavocytochrome c-sulfide dehydrogenase (FCSD), having 49% identity. These data allowed isolation of a 5.5-kb subgenomic clone which contains the cytochrome gene and an adjacent flavoprotein gene as in other species which have an FCSD. The cytochrome subunit has a signal peptide with a normal cleavage site, but the flavoprotein subunit has a signal sequence which suggests that the mature protein has an N-terminal cysteine, characteristic of a diacyl glycerol-modified lipoprotein. The membrane localization of FCSD was confirmed by Western blotting with antibodies raised against Chromatium vinosum FCSD. When aligned according to the three-dimensional structure of Chromatium FCSD, all but one of the side chains near the flavin are conserved. These include the Cys 42 flavin adenine dinucleotide binding site; the Cys 161-Cys 337 disulfide; Glu 167, which modulates the reactivity with sulfite; and aromatic residues which may function as charge transfer acceptors from the flavin-sulfite adduct (C. vinosum numbering). The genetic context of FCSD is different from that in other species in that flanking genes are not conserved. The transcript is only large enough to encode the two FCSD subunits. Furthermore, Northern hybridization showed that the production of E. vacuolata FCSD mRNA is regulated by sulfide. All cultures that contained sulfide in the medium had elevated levels of FCSD RNA compared with cells grown on organics (acetate, malate, or succinate) or thiosulfate alone, consistent with the role of FCSD in sulfide oxidation. PMID- 10809688 TI - A complex insertion sequence cluster at a point of interaction between the linear plasmid SCP1 and the linear chromosome of Streptomyces coelicolor A3(2). AB - The giant linear plasmid SCP1 can integrate into the central region of the linear chromosome of Streptomyces coelicolor A3(2). Nucleotide sequence analysis around the target site for SCP1 integration in strain M145 identified a total of five copies of four insertion sequences (ISs) in a 6.5-kb DNA stretch. Three of the four (IS468, IS469, and IS470) are new IS elements, and the other is IS466. All of these elements contain one open reading frame which encodes a transposase-like protein. Two copies of IS468 (IS468A and -B) are tandemly aligned at the left end of the cluster. Following these, IS469 and IS466 are located in a tail-to-tail orientation with 69.3% identity to each other. IS470 is located at the right end of the cluster. The activities of IS466 and IS468 were demonstrated by transposition experiments and sequence comparison of several copies, respectively. PMID- 10809689 TI - Proteolysis of bacteriophage lambda CII by Escherichia coli FtsH (HflB). AB - FtsH (HflB) is a conserved, highly specific, ATP-dependent protease for which a number of substrates are known. The enzyme participates in the phage lambda lysis lysogeny decision by degrading the lambda CII transcriptional activator and by its response to inhibition by the lambda CIII gene product. In order to gain further insight into the mechanism of the enzymatic activity of FtsH (HflB), we identified the peptides generated following proteolysis of the phage lambda CII protein. It was found that FtsH (HflB) acts as an endopeptidase degrading CII into small peptides with limited amino acid specificity at the cleavage site. beta-Casein, an unstructured substrate, is also degraded by FtsH (HflB), suggesting that protein structure may play a minor role in determining the products of proteolysis. The majority of the peptides produced were 13 to 20 residues long. PMID- 10809690 TI - Structural elements required for replication and incompatibility of the Rhizobium etli symbiotic plasmid. AB - The symbiotic plasmid of Rhizobium etli CE3 belongs to the RepABC family of plasmid replicons. This family is characterized by the presence of three conserved genes, repA, repB, and repC, encoded by the same DNA strand. A long intergenic sequence (igs) between repB and repC is also conserved in all members of the plasmid family. In this paper we demonstrate that (i) the repABC genes are organized in an operon; (ii) the RepC product is essential for replication; (iii) RepA and RepB products participate in plasmid segregation and in the regulation of plasmid copy number; (iv) there are two cis-acting incompatibility regions, one located in the igs (incalpha) and the other downstream of repC (incbeta) (the former is essential for replication); and (v) RepA is a trans-acting incompatibility factor. We suggest that incalpha is a cis-acting site required for plasmid partitioning and that the origin of replication lies within incbeta. PMID- 10809691 TI - Sequence diversity of Pseudomonas aeruginosa: impact on population structure and genome evolution. AB - Comparative sequencing of Pseudomonas aeruginosa genes oriC, citS, ampC, oprI, fliC, and pilA in 19 environmental and clinical isolates revealed the sequence diversity to be about 1 order of magnitude lower than in comparable housekeeping genes of Salmonella. In contrast to the low nucleotide substitution rate, the frequency of recombination among different P. aeruginosa genotypes was high, leading to the random association of alleles. The P. aeruginosa population consists of equivalent genotypes that form a net-like population structure. However, each genotype represents a cluster of closely related strains which retain their sequence signature in the conserved gene pool and carry a set of genotype-specific DNA blocks. The codon adaptation index, a quantitative measure of synonymous codon bias of genes, was found to be consistently high in the P. aeruginosa genome irrespective of the metabolic category and the abundance of the encoded gene product. Such uniformly high codon adaptation indices of 0.55 to 0.85 fit the ubiquitous lifestyle of P. aeruginosa. PMID- 10809692 TI - Cloning and expression of ntnD, encoding a novel NAD(P)(+)-independent 4 nitrobenzyl alcohol dehydrogenase from Pseudomonas sp. Strain TW3. AB - Pseudomonas sp. strain TW3 is able to metabolize 4-nitrotoluene to 4 nitrobenzoate and toluene to benzoate aerobically via a route analogous to the upper pathway of the TOL plasmids. We report the cloning and characterization of a benzyl alcohol dehydrogenase gene (ntnD) which encodes the enzyme for the catabolism of 4-nitrobenzyl alcohol and benzyl alcohol to 4-nitrobenzaldehyde and benzaldehyde, respectively. The gene is located downstream of the previously reported ntn gene cluster. NtnD bears no similarity to the analogous TOL plasmid XylB (benzyl alcohol dehydrogenase) protein either in its biochemistry, being NAD(P)(+) independent and requiring assay via dye-linked electron transfer, or in its deduced amino acid sequence. It does, however, have significant similarity in its amino acid sequence to other NAD(P)(+)-independent alcohol dehydrogenases and contains signature patterns characteristic of type III flavin adenine dinucleotide-dependent alcohol oxidases. Reverse transcription-PCR demonstrated that ntnD is transcribed during growth on 4-nitrotoluene, although apparently not as part of the same transcript as the other ntn genes. The substrate specificity of the enzyme expressed from the cloned and overexpressed gene was similar to the activity expressed from strain TW3 grown on 4-nitrotoluene, providing evidence that ntnD is the previously unidentified gene in the pathway of 4-nitrotoluene catabolism. Examination of the 14.8-kb region around the ntn genes suggests that one or more recombination events have been involved in the formation of their current organization. PMID- 10809693 TI - Interplay between efflux pumps may provide either additive or multiplicative effects on drug resistance. AB - The effects of simultaneous expression of several efflux pumps on antibiotic resistance were investigated in Escherichia coli and Pseudomonas aeruginosa. Several combinations of efflux pumps have been studied: (i) simultaneous expression of a single-component efflux pump, which exports antibiotics into the periplasm, in combination with a multicomponent efflux pump that accomplishes efflux directly into the external medium; (ii) simultaneous expression of two single-component pumps; and (iii) simultaneous expression of two multicomponent pumps. It was found that when efflux pumps of different structural types were combined in the same cell (the first case), the observed antibiotic resistance was much higher than that conferred by each of the pumps expressed singly. Simultaneous expression of pairs of single-component or multicomponent efflux pumps (the second and third cases) did not produce strong increases in antibiotic resistance. PMID- 10809694 TI - lon incompatibility associated with mutations causing SOS induction: null uvrD alleles induce an SOS response in Escherichia coli. AB - The uvrD gene in Escherichia coli encodes a 720-amino-acid 3'-5' DNA helicase which, although nonessential for viability, is required for methyl-directed mismatch repair and nucleotide excision repair and furthermore is believed to participate in recombination and DNA replication. We have shown in this study that null mutations in uvrD are incompatible with lon, the incompatibility being a consequence of the chronic induction of SOS in uvrD strains and the resultant accumulation of the cell septation inhibitor SulA (which is a normal target for degradation by Lon protease). uvrD-lon incompatibility was suppressed by sulA, lexA3(Ind(-)), or recA (Def) mutations. Other mutations, such as priA, dam, polA, and dnaQ (mutD) mutations, which lead to persistent SOS induction, were also lon incompatible. SOS induction was not observed in uvrC and mutH (or mutS) mutants defective, respectively, in excision repair and mismatch repair. Nor was uvrD mediated SOS induction abolished by mutations in genes that affect mismatch repair (mutH), excision repair (uvrC), or recombination (recB and recF). These data suggest that SOS induction in uvrD mutants is not a consequence of defects in these three pathways. We propose that the UvrD helicase participates in DNA replication to unwind secondary structures on the lagging strand immediately behind the progressing replication fork, and that it is the absence of this function which contributes to SOS induction in uvrD strains. PMID- 10809695 TI - In Saccharomyces cerevisiae, expression of arginine catabolic genes CAR1 and CAR2 in response to exogenous nitrogen availability is mediated by the Ume6 (CargRI) Sin3 (CargRII)-Rpd3 (CargRIII) complex. AB - The products of three genes named CARGRI, CARGRII, and CARGRIII were shown to repress the expression of CAR1 and CAR2 genes, involved in arginine catabolism. CARGRI is identical to UME6 and encodes a regulator of early meiotic genes. In this work we identify CARGRII as SIN3 and CARGRIII as RPD3. The associated gene products are components of a high-molecular-weight complex with histone deacetylase activity and are recruited by Ume6 to promoters containing a URS1 sequence. Sap30, another component of this complex, is also required to repress CAR1 expression. This histone deacetylase complex prevents the synthesis of the two arginine catabolic enzymes, arginase (CAR1) and ornithine transaminase (CAR2), as long as exogenous nitrogen is available. Upon nitrogen depletion, repression at URS1 is released and Ume6 interacts with ArgRI and ArgRII, two proteins involved in arginine-dependent activation of CAR1 and CAR2, leading to high levels of the two catabolic enzymes despite a low cytosolic arginine pool. Our data also show that the deletion of the UME6 gene impairs cell growth more strongly than the deletion of the SIN3 or RPD3 gene, especially in the Sigma1278b background. PMID- 10809696 TI - Mutually exclusive utilization of P(R) and P(RM) promoters in bacteriophage 434 O(R). AB - Establishment and maintenance of a lysogen of the lambdoid bacteriophage 434 require that the 434 repressor both activate transcription from the P(RM) promoter and repress transcription from the divergent P(R) promoter. Several lines of evidence indicate that the 434 repressor activates initiation of P(RM) transcription by occupying a binding site adjacent to the P(RM) promoter and directly contacting RNA polymerase. The overlapping architecture of the P(RM) and P(R) promoters suggests that an RNA polymerase bound at P(R) may repress P(RM) transcription initiation. Hence, part of the stimulatory effect of the 434 repressor may be relief of interference between RNA polymerase binding to the P(RM) promoter and to the P(R) promoter. Consistent with this proposal, we show that the repressor cannot activate P(RM) transcription if RNA polymerase binds at P(R) prior to addition of the 434 repressor. However, unlike the findings with the related lambda phage, formation of RNA polymerase promoter complexes at P(RM) and at P(R) apparently are mutually exclusive. We find that the RNA polymerase mediated inhibition of repressor-stimulated P(RM) transcription requires the presence of an open complex at P(R). Taken together, these results indicate that establishment of an open complex at P(R) directly prevents formation of an RNA polymerase-P(RM) complex. PMID- 10809697 TI - Physical mapping of bchG, orf427, and orf177 in the photosynthesis gene cluster of Rhodobacter sphaeroides: functional assignment of the bacteriochlorophyll synthetase gene. AB - The purple photosynthetic bacterium Rhodobacter sphaeroides has within its genome a cluster of photosynthesis-related genes approximately 41 kb in length. In an attempt to identify genes involved in the terminal esterification stage of bacteriochlorophyll biosynthesis, a previously uncharacterized 5-kb region of this cluster was sequenced. Four open reading frames (ORFs) were identified, and each was analyzed by transposon mutagenesis. The product of one of these ORFs, bchG, shows close homologies with (bacterio)chlorophyll synthetases, and mutants in this gene were found to accumulate bacteriopheophorbide, the metal-free derivative of the bacteriochlorophyll precursor bacteriochlorophyllide, suggesting that bchG is responsible for the esterification of bacteriochlorophyllide with an alcohol moiety. This assignment of function to bchG was verified by the performance of assays demonstrating the ability of BchG protein, heterologously synthesized in Escherichia coli, to esterify bacteriochlorophyllide with geranylgeranyl pyrophosphate in vitro, thereby generating bacteriochlorophyll. This step is pivotal to the assembly of a functional photosystem in R. sphaeroides, a model organism for the study of structure-function relationships in photosynthesis. A second gene, orf177, is a member of a large family of isopentenyl diphosphate isomerases, while sequence homologies suggest that a third gene, orf427, may encode an assembly factor for photosynthetic complexes. The function of the remaining ORF, bchP, is the subject of a separate paper (H. Addlesee and C. N. Hunter, J. Bacteriol. 181:7248-7255, 1999). An operonal arrangement of the genes is proposed. PMID- 10809698 TI - Yersinia enterocolitica TyeA, an intracellular regulator of the type III machinery, is required for specific targeting of YopE, YopH, YopM, and YopN into the cytosol of eukaryotic cells. AB - Pathogenic Yersinia species employ type III machines to target effector Yops into the cytosol of eukaryotic cells. Yersinia tyeA mutants are thought to be defective in the targeting of YopE and YopH without affecting the injection of YopM, YopN, YopO, YopP, and YopT into the cytosol of eukaryotic cells. One model suggests that TyeA may form a tether between YopN (LcrE) and YopD on the bacterial surface, a structure that may translocate YopE and YopH across the plasma membrane of eukaryotic cells (M. Iriarte, M. P. Sory, A. Boland, A. P. Boyd, S. D. Mills, I. Lambermont, and G. R. Cornelis, EMBO J. 17:1907-1918, 1998). We have examined the injection of Yop proteins by tyeA mutant yersiniae with the digitonin fractionation technique. We find that tyeA mutant yersiniae not only secreted YopE, YopH, YopM, and YopN into the extracellular medium but also targeted these polypeptides into the cytosol of HeLa cells. Protease protection, cell fractionation, and affinity purification experiments suggest that TyeA is located intracellularly and binds to YopN or YopD. We propose a model whereby TyeA functions as a negative regulator of the type III targeting pathway in the cytoplasm of yersiniae, presumably by preventing the export of YopN. PMID- 10809699 TI - Initiation and termination of DNA transfer during conjugation of IncI1 plasmid R64: roles of two sets of inverted repeat sequences within oriT in termination of R64 transfer. AB - Intercellular transfer of plasmid DNA during bacterial conjugation initiates and terminates at a specific origin of transfer, oriT. We have investigated the oriT structure of conjugative plasmid R64 with regard to the initiation and termination of DNA transfer. Using recombinant plasmids containing two tandemly repeated R64 oriT sequences with or without mutations, the subregions required for initiation and termination were determined by examining conjugation-mediated deletion between the repeated oriTs. The oriT subregion required for initiation was found to be identical to the 44-bp oriT core sequence consisting of two units, the conserved nick region sequence and the 17-bp repeat A sequence, that are recognized by R64 relaxosome proteins NikB and NikA, respectively. In contrast, the nick region sequence and two sets of inverted repeat sequences within the 92-bp minimal oriT sequence were required for efficient termination. Mutant repeat A sequences lacking NikA-binding ability were found to be sufficient for termination, suggesting that the inverted repeat structures are involved in the termination process. A duplication of the DNA segment between the repeated oriTs was also found after mobilization of the plasmid carrying initiation-deficient but termination-proficient oriT and initiation-proficient but termination-deficient oriT, suggesting that the 3' terminus of the transferred strand is elongated by rolling-circle-DNA synthesis. PMID- 10809700 TI - Identification, cloning, and initial characterization of rot, a locus encoding a regulator of virulence factor expression in Staphylococcus aureus. AB - A chromosomal insertion of transposon Tn917 partially restores the expression of protease and alpha-toxin activities to PM466, a genetically defined agr-null derivative of the wild-type Staphylococcus aureus strain RN6390. In co transduction experiments, transposon-encoded erythromycin resistance and a protease- and alpha-toxin-positive phenotype are transferred at high frequency from mutant strains to agr-null strains of S. aureus. Southern analysis of chromosomal DNA and sequence analysis of DNA flanking the Tn917 insertion site in mutant strains revealed that the transposon interrupted a 498-bp open reading frame (ORF). Similarity searches using a conceptual translation of the ORF identified a region of homology to the known staphylococcal global regulators AgrA and SarA. To verify that the mutant allele conferred the observed phenotype, a wild-type allele of the mutant gene was introduced into the genome of a mutant strain by homologous recombination. The resulting isolates had a restored agr null phenotype. Virulence factor gene expression in mutant, restored mutant, and wild-type strains was quantified by measuring alpha-toxin activity in culture supernatant fluids and by Northern analysis of the alpha-toxin transcript. We named this ORF rot (for repressor of toxins) (GenBank accession no. AF189239) because of the activity associated with rot::Tn917 mutant strains. PMID- 10809701 TI - Another unusual type of citric acid cycle enzyme in Helicobacter pylori: the malate:quinone oxidoreductase. AB - The only enzyme of the citric acid cycle for which no open reading frame (ORF) was found in the Helicobacter pylori genome is the NAD-dependent malate dehydrogenase. Here, it is shown that in this organism the oxidation of malate to oxaloacetate is catalyzed by a malate:quinone oxidoreductase (MQO). This flavin adenine dinucleotide-dependent membrane-associated enzyme donates electrons to quinones of the electron transfer chain. Similar to succinate dehydrogenase, it is part of both the electron transfer chain and the citric acid cycle. MQO activity was demonstrated in isolated membranes of H. pylori. The enzyme is encoded by the ORF HP0086, which is shown by the fact that expression of the HP0086 sequence from a plasmid induces high MQO activity in mqo deletion mutants of Escherichia coli or Corynebacterium glutamicum. Furthermore, this plasmid was able to complement the phenotype of the C. glutamicum mqo deletion mutant. Interestingly, the protein predicted to be encoded by this ORF is only distantly related to known or postulated MQO sequences from other bacteria. The presence of an MQO shown here and the previously demonstrated presence of a 2 ketoglutarate:ferredoxin oxidoreductase and a succinyl-coenzyme A (CoA):acetoacetyl-CoA transferase indicate that H. pylori possesses a complete citric acid cycle, but one which deviates from the standard textbook example in three steps. PMID- 10809702 TI - Differences in genotypes of Helicobacter pylori from different human populations. AB - DNA motifs at several informative loci in more than 500 strains of Helicobacter pylori from five continents were studied by PCR and sequencing to gain insights into the evolution of this gastric pathogen. Five types of deletion, insertion, and substitution motifs were found at the right end of the H. pylori cag pathogenicity island. Of the three most common motifs, type I predominated in Spaniards, native Peruvians, and Guatemalan Ladinos (mixed Amerindian-European ancestry) and also in native Africans and U.S. residents; type II predominated among Japanese and Chinese; and type III predominated in Indians from Calcutta. Sequences in the cagA gene and in vacAm1 type alleles of the vacuolating cytotoxin gene (vacA) of strains from native Peruvians were also more like those from Spaniards than those from Asians. These indications of relatedness of Latin American and Spanish strains, despite the closer genetic relatedness of Amerindian and Asian people themselves, lead us to suggest that H. pylori may have been brought to the New World by European conquerors and colonists about 500 years ago. This thinking, in turn, suggests that H. pylori infection might have become widespread in people quite recently in human evolution. PMID- 10809703 TI - Distinctiveness of genotypes of Helicobacter pylori in Calcutta, India. AB - The genotypes of 78 strains of Helicobacter pylori from Calcutta, India (55 from ulcer patients and 23 from more-benign infections), were studied, with a focus on putative virulence genes and neutral DNA markers that were likely to be phylogenetically informative. PCR tests indicated that 80 to 90% of Calcutta strains carried the cag pathogenicity island (PAI) and potentially toxigenic vacAs1 alleles of the vacuolating cytotoxin gene (vacA), independent of disease status. This was higher than in the West (where cag PAI(+) vacAs1 genotypes are disease associated) but lower than in east Asia. The iceA2 gene was weakly disease associated in Calcutta, whereas in the West the alternative but unrelated iceA1 gene at the same locus is weakly disease associated. DNA sequence motifs of vacAm1 (middle region) alleles formed a cluster that was distinct from those of east Asia and the West, whereas the cagA sequences of Calcutta and Western strains were closely related. An internal deletion found in 20% of Calcutta iceA1 genes was not seen in any of approximately 200 strains studied from other geographic regions and thus seemed to be unique to this H. pylori population. Two mobile DNAs that were rare in east Asian strains were also common in Calcutta. About 90% of Calcutta strains were metronidazole resistant. These findings support the idea that H. pylori gene pools differ regionally and emphasize the potential importance of studies of Indian and other non-Western H. pylori populations in developing a global understanding of this gastric pathogen and associated disease. PMID- 10809704 TI - Differential activation of the tcpPH promoter by AphB determines biotype specificity of virulence gene expression in Vibrio cholerae. AB - Vibrio cholerae strains of the classical biotype express the genes encoding cholera toxin (CT) and toxin-coregulated pilus (TCP) under a variety of environmental conditions in vitro, whereas El Tor biotype strains express these genes only under specialized culture conditions. We show here that a single base pair difference at positions -65 and -66 of the classical and El Tor tcpPH promoters, respectively, is responsible for the differential regulation of virulence gene expression in these two disease-causing biotypes. Analysis of tcpP lacZ fusions in both V. cholerae and Escherichia coli indicated that transcriptional activation of the El Tor tcpPH promoter by the LysR regulator AphB was significantly reduced relative to that of the classical promoter. Reciprocal exchange of the tcpPH promoter between the two biotypes in V. cholerae showed that the ability to activate the transcription of tcpPH is not dependent on the biotype of the strain per se but on the tcpPH promoter itself. Classical and El Tor tcpP-lacZ promoter chimeras in E. coli localized the region responsible for the differential activation of tcpPH by AphB to within 75 bp of the transcriptional start site. Individual base-pair changes within this region showed that the presence of either an A or a G at position -65 or -66 conferred the classical or El Tor, respectively, pattern of tcpPH activation by AphB. Reciprocal exchange of these base pairs between biotypes in V. cholerae switched the biotype-specific pattern of expression of tcpPH as well as the production of CT and TCP in response to environmental stimuli. PMID- 10809705 TI - Branched-chain alpha-keto acid catabolism via the gene products of the bkd operon in Enterococcus faecalis: a new, secreted metabolite serving as a temporary redox sink. AB - Recently the bkd gene cluster from Enterococcus faecalis was sequenced, and it was shown that the gene products constitute a pathway for the catabolism of branched-chain alpha-keto acids. We have now investigated the regulation and physiological role of this pathway. Primer extension analysis identified the presence of a single promoter upstream of the bkd gene cluster. Furthermore, a putative catabolite-responsive element was identified in the promoter region, indicative of catabolite repression. Consistent with this was the observation that expression of the bkd gene cluster is repressed in the presence of glucose, fructose, and lactose. It is proposed that the conversion of the branched-chain alpha-keto acids to the corresponding free acids results in the formation of ATP via substrate level phosphorylation. The utilization of the alpha-keto acids resulted in a marked increase of biomass, equivalent to a net production of 0.5 mol of ATP per mol of alpha-keto acid metabolized. The pathway was active under aerobic as well as anaerobic conditions. However, under anaerobic conditions the presence of a suitable electron acceptor to regenerate NAD(+) from the NADH produced by the branched-chain alpha-keto acid dehydrogenase complex was required for complete conversion of alpha-ketoisocaproate. Interestingly, during the conversion of the branched-chain alpha-keto acids an intermediate was always detected extracellularly. With alpha-ketoisocaproic acid as the substrate this intermediate was tentatively identified as 1, 1-dihydroxy-4-methyl-2-pentanone. This reduced form of alpha-ketoisocaproic acid was found to serve as a temporary redox sink. PMID- 10809706 TI - Analysis of genes encoding an alternative nitrogenase in the archaeon Methanosarcina barkeri 227. AB - Methanosarcina barkeri 227 possesses two clusters of genes potentially encoding nitrogenases. We have previously demonstrated that one cluster, called nif2, is expressed under molybdenum (Mo)-sufficient conditions, and the deduced amino acid sequences for nitrogenase structural genes in that cluster most closely resemble those for the Mo nitrogenase of the gram-positive eubacterium Clostridium pasteurianum. The previously cloned nifH1 from M. barkeri shows phylogenetic relationships with genes encoding components of eubacterial Mo-independent eubacterial alternative nitrogenases and other methanogen nitrogenases. In this study, we cloned and sequenced nifD1 and part of nifK1 from M. barkeri 227. The deduced amino acid sequence encoded by nifD1 from M. barkeri showed great similarity with vnfD gene products from vanadium (V) nitrogenases, with an 80% identity at the amino acid level with the vnfD gene product from Anabaena variabilis. Moreover, there was a small open reading frame located between nifD1 and nifK1 with clear homology to vnfG, a hallmark of eubacterial alternative nitrogenases. Stimulation of diazotrophic growth of M. barkeri 227 by V in the absence of Mo was demonstrated. The unusual complement of nif genes in M. barkeri 227, with one cluster resembling that from a gram-positive eubacterium and the other resembling a eubacterial V nitrogenase gene cluster, suggests horizontal genetic transfer of those genes. PMID- 10809707 TI - Characterization of the streptococcal C5a peptidase using a C5a-green fluorescent protein fusion protein substrate. AB - A glutathione-S-transferase (GST)-C5a-green fluorescent protein (GFP) fusion protein was designed for use as a substrate for the streptococcal C5a peptidase (SCPA). The substrate was immobilized on a glutathione-Sepharose affinity matrix and used to measure wild-type SCPA activity in the range of 0.8 to 800 nM. The results of the assay demonstrated that SCPA is highly heat stable and has optimal activity on the synthetic substrate at or above pH 8.0. SCPA activity was unaffected by 0.1 to 10 mM Ca(2+), Mg(2+), and Mn(2+) but was inhibited by the same concentrations of Zn(2+). The assay shows high sensitivity to ionic strength; NaCl inhibits SCPA cleavage of GST-C5a-GFP in a dose-dependent manner. Based on previously published computer homology modeling, four substitutions were introduced into the putative active site of SCPA: Asp(130)-Ala, His(193)-Ala, Asn(295)-Ala, and Ser(512)-Ala. All four mutant proteins had over 1,000-fold less proteolytic activity on C5a in vitro, as determined both by the GFP assay described here and by a polymorphonuclear cell adherence assay. In addition, recombinant SCPA1 and SCPA49, from two distinct lineages of Streptococcus pyogenes (group A streptococci), and recombinant SCPB, from Streptococcus agalactiae (group B streptococci), were compared in the GFP assay. The three enzymes had similar activities, all cleaving approximately 6 mol of C5a mmol of SCP(-1) liter(-1) min(-1). PMID- 10809708 TI - The clp proteases of Bacillus subtilis are directly involved in degradation of misfolded proteins. AB - The presence of the heat stress response-related ATPases ClpC and ClpX or the peptidase ClpP in the cell is crucial for tolerance of many forms of stress in Bacillus subtilis. Assays for detection of defects in protein degradation suggest that ClpC, ClpP, and ClpX participate directly in overall proteolysis of misfolded proteins. Turnover rates for abnormal puromycyl peptides are significantly decreased in clpC, clpP, and clpX mutant cells. Electron-dense aggregates, most likely due to the accumulation of misfolded proteins, were noticed in studies of ultrathin cryosections in clpC and clpP mutant cells even under nonstress conditions. In contrast, in the wild type or clpX mutants such aggregates could only be observed after heat shock. This phenomenon supports the assumption that clpC and clpP mutants are deficient in the ability to solubilize or degrade damaged and aggregated proteins, the accumulation of which is toxic for the cell. By using immunogold labeling with antibodies raised against ClpC, ClpP, and ClpX, the Clp proteins were localized in these aggregates, showing that the Clp proteins act at this level in vivo. PMID- 10809709 TI - Mutational analysis of the sbo-alb locus of Bacillus subtilis: identification of genes required for subtilosin production and immunity. AB - The Bacillus subtilis 168 derivative JH642 produces a bacteriocin, subtilosin, which possesses activity against Listeria monocytogenes. Inspection of the amino acid sequence of the presubtilosin polypeptide encoded by the gene sboA and sequence data from analysis of mature subtilosin indicate that the precursor subtilosin peptide undergoes several unique and unusual chemical modifications during its maturation process. The genes of the sbo-alb operon are believed to function in the synthesis and maturation of subtilosin. Nonpolar mutations introduced into each of the alb genes resulted in loss or reduction of subtilosin production. sboA, albA, and albF mutants showed no antilisterial activity, indicating that the products of these genes are critical for the production of active subtilosin. Mutations in albB, -C, and -D resulted in reduction of antilisterial activity and decreased immunity to subtilosin, particularly under anaerobic conditions. A new gene, sboX, encoding another bacteriocin-like product was discovered residing in a sequence overlapping the coding region of sboA. Construction of an sboX-lacZ translational fusion and analysis of its expression indicate that sboX is induced in stationary phase of anaerobic cultures of JH642. An in-frame deletion of the sboX coding sequence did not affect the antilisterial activity or production of or immunity to subtilosin. The results of this investigation show that the sbo-alb genes are required for the mechanisms of subtilosin synthesis and immunity. PMID- 10809710 TI - Dual control of sbo-alb operon expression by the Spo0 and ResDE systems of signal transduction under anaerobic conditions in Bacillus subtilis. AB - The Bacillus subtilis sbo-alb operon contains sboA, the structural gene for the bacteriocin subtilosin, and the alb genes required for subtilosin production. Transcription from the sbo-alb promoter is highly induced by oxygen limitation. The transcriptional regulation of the sbo-alb operon is under dual control involving the transition state regulator AbrB and the two-component regulatory proteins ResD and ResE. PMID- 10809711 TI - Natural diversity in the N terminus of the mature vacuolating cytotoxin of Helicobacter pylori determines cytotoxin activity. AB - Naturally occurring noncytotoxic vacA type s2 strains of Helicobacter pylori have a 12-residue extension to the vacuolating cytotoxin (VacA) compared with cytotoxic type s1 strains. We show that adding the region encoding this extension to type s1 vacA completely abolishes vacuolating cytotoxin activity but has no effect on VacA production. PMID- 10809712 TI - Identification of a small RNA within the pdh gene cluster of Mycoplasma pneumoniae and Mycoplasma genitalium. AB - A highly abundant and heterogeneous small RNA about 205 to 210 bases long named MP200 RNA has been identified in Mycoplasma pneumoniae. It was localized on the genome within a 319-bp-long intergenic space of the pyruvate dehydrogenase (pdh) gene cluster. A database search at the DNA level revealed the highest similarity to a sequence located within the pdh gene cluster of Mycoplasma genitalium that was also shown to be transcribed into two abundant, but smaller RNAs than the ones in Mycoplasma pneumoniae. The RNAs from both M. pneumoniae and M. genitalium have the potential to code for cysteine-rich 29- and 23-amino-acid-long peptides, but so far, these peptides have not been identified experimentally in bacterial protein extracts. PMID- 10809713 TI - Restart of exponential growth of cold-shocked Yersinia enterocolitica occurs after down-regulation of cspA1/A2 mRNA. AB - The cellular content of major cold shock protein (MCSP) mRNA transcribed from the tandem gene duplication cspA1/A2 and growth of Yersinia enterocolitica were compared when exponentially growing cultures of this bacterium were cold shocked from 30 to 20, 15, 10, 5, or 0 degrees C, respectively. A clear correlation between the time point when exponential growth resumes after cold shock and the degradation of cspA1/A2 mRNA was found. A polynucleotide phosphorylase-deficient mutant was unable to degrade cspA1/A2 mRNA properly and showed a delay, as well as a lower rate, of growth after cold shock. For this mutant, a correlation between decreasing cspA1/A2 mRNA and restart of growth after cold shock was also observed. For both wild-type and mutant cells, no correlation of restart of growth with the cellular content of MCSPs was found. We suggest that, after synthesis of cold shock proteins and cold adaptation of the cells, MCSP mRNAs must be degraded; otherwise, they trap ribosomes, prevent translation of bulk mRNA, and thus inhibit growth of this bacterium at low temperatures. PMID- 10809714 TI - Transformation of Rickettsia prowazekii to erythromycin resistance encoded by the Escherichia coli ereB gene. AB - Rickettsia prowazekii, the etiologic agent of epidemic typhus, is an obligate, intracytoplasmic, parasitic bacterium. Recently, the transformation of this bacterium via electroporation has been reported. However, in these studies identification of transformants was dependent upon either selection of an R. prowazekii rpoB chromosomal mutation imparting rifampin resistance or expression of the green fluorescent protein and flow cytometric analysis. In this paper we describe the expression in R. prowazekii of the Escherichia coli ereB gene. This gene codes for an erythromycin esterase that cleaves erythromycin. To the best of our knowledge, this is the first report of the expression of a nonrickettsial, antibiotic-selectable gene in R. prowazekii. The availability of a positive selection for rickettsial transformants is an important step in the characterization of genetic analysis systems in the rickettsiae. PMID- 10809715 TI - Organization, structure, and variability of the rRNA operon of the Whipple's disease bacterium (Tropheryma whippelii). AB - Whipple's disease is a systemic disorder associated with a cultivation-resistant, poorly characterized actinomycete, Tropheryma whippelii. We determined a nearly complete rRNA operon sequence of T. whippelii from specimens from 3 patients with Whipple's disease, as well as partial operon sequences from 43 patients. Variability was observed in the 16S-23S rRNA spacer sequences, leading to the description of five distinct sequence types. One specimen contained two spacer sequence types, raising the possibility of a double infection. Secondary structure models for the primary rRNA transcript and mature rRNAs revealed rare or unique features. PMID- 10809716 TI - Secretion of the Caulobacter crescentus S-layer protein: further localization of the C-terminal secretion signal and its use for secretion of recombinant proteins. AB - The secretion signal of the Caulobacter crescentus S-layer protein (RsaA) was localized to the C-terminal 82 amino acids of the molecule. Protein yield studies showed that 336 or 242 C-terminal residues of RsaA mediated secretion of >50 mg of a cellulase passenger protein per liter to the culture fluids. PMID- 10809717 TI - Characterization of the relA gene of Porphyromonas gingivalis. AB - Based upon the nucleotide sequence of the relA gene from Escherichia coli, a gene fragment corresponding to the homologous gene from the pathogenic oral bacterium Porphyromonas gingivalis 381 was isolated by PCR and utilized to construct a relA mutant. The mutant, KS7, was defective in ribosome-mediated ppGpp formation and also in the stringent response. PMID- 10809718 TI - Evidence that SpoIVFB is a novel type of membrane metalloprotease governing intercompartmental communication during Bacillus subtilis sporulation. AB - Processing of pro-sigma(K) in the mother cell compartment of sporulating Bacillus subtilis involves SpoIVFB and is governed by a signal from the forespore. SpoIVFB has an HEXXH motif characteristic of metalloproteases embedded in one of its transmembrane segments. Several conservative single amino acid changes in the HEXXH motif abolished function. However, changing the glutamic acid residue to aspartic acid, or changing the isoleucine residue that precedes the motif to proline, permitted SpoIVFB function. Only one other putative metalloprotease, site 2 protease has been shown to tolerate aspartic acid rather than glutamic acid in its HEXXH sequence. Site 2 protease and SpoIVFB share a second region of similarity with a family of putative membrane metalloproteases. A conservative change in this region of SpoIVFB abolished function. Interestingly, SpoIVFA increased the accumulation of certain mutant SpoIVFB proteins but was unnecessary for accumulation of wild-type SpoIVFB. PMID- 10809719 TI - Target site selection by Tn7: attTn7 transcription and target activity. AB - The bacterial transposon Tn7 inserts at high frequency into a specific site called attTn7, which is present in the chromosomes of many bacteria. We show here that transcription of a nearby gene, glmS, decreases the frequency of Tn7 insertion into attTn7, thus providing a link between Tn7 transposition and host cell metabolism. PMID- 10809720 TI - A slow-motility phenotype caused by substitutions at residue Asp31 in the PomA channel component of a sodium-driven flagellar motor. AB - PomA is thought to be a component of the ion channel in the sodium-driven polar flagellar motor of Vibrio alginolyticus. We have found that some cysteine substitutions in the periplasmic region of PomA result in a slow-motility phenotype, in which swarming and swimming speeds are reduced even in the presence of high concentrations of NaCl. Most of the mutants showed a sodium ion dependence similar to that of the wild type but with significantly reduced motility at all sodium ion concentrations. By contrast, motility of the D31C mutant showed a sharp dependence on NaCl concentration, with a threshold at 38 mM. The motor of the D31C mutant rotates stably, as monitored by laser dark-field microscopy, suggesting that the mutant PomA protein is assembled normally into the motor complex. Mutational studies of Asp31 suggest that, although this residue is not essential for motor rotation, a negative charge at this position contributes to optimal speed and/or efficiency of the motor. PMID- 10809721 TI - Asp-170 is crucial for the redox properties of vanillyl-alcohol oxidase. AB - Vanillyl-alcohol oxidase is a flavoprotein containing a covalent flavin that catalyzes the oxidation of 4-(methoxymethyl)phenol to 4-hydroxybenzaldehyde. The reaction proceeds through the formation of a p-quinone methide intermediate, after which, water addition takes place. Asp-170, located near the N5-atom of the flavin, has been proposed to act as an active site base. To test this hypothesis, we have addressed the properties of D170E, D170S, D170A, and D170N variants. Spectral and fluorescence analysis, together with the crystal structure of D170S, suggests that the Asp-170 replacements do not induce major structural changes. However, in D170A and D170N, 50 and 100%, respectively, of the flavin is non covalently bound. Kinetic characterization of the vanillyl-alcohol oxidase variants revealed that Asp-170 is required for catalysis. D170E is 50-fold less active, and the other Asp-170 variants are about 10(3)-fold less active than wild type enzyme. Impaired catalysis of the Asp-170 variants is caused by slow flavin reduction. Furthermore, the mutant proteins have lost the capability of forming a stable complex between reduced enzyme and the p-quinone methide intermediate. The redox midpoint potentials in D170E (+6 mV) and D170S (-91 mV) are considerably decreased compared with wild type vanillyl-alcohol oxidase (+55 mV). This supports the idea that Asp-170 interacts with the protonated N5-atom of the reduced cofactor, thus increasing the FAD redox potential. Taken together, we conclude that Asp-170 is involved in the process of autocatalytic flavinylation and is crucial for efficient redox catalysis. PMID- 10809722 TI - Biochemical characterization of the catalytic domain of human matrix metalloproteinase 19. Evidence for a role as a potent basement membrane degrading enzyme. AB - We have recently cloned MMP-19, a novel matrix metalloproteinase, which, due to unique structural features, was proposed to represent the first member of a new MMP subfamily (Pendas, A. M., Knauper, V. , Puente, X. S., Llano, E., Mattei, M. G., Apte, S., Murphy, G., and Lopez-Otin, C. (1997) J. Biol. Chem. 272, 4281 4286). A recombinant COOH-terminal deletion mutant of MMP-19 (proDelta(260 508)MMP-19), comprising the propeptide and the catalytic domain, was expressed in Escherichia coli, refolded, and purified. Interestingly, we found that proDelta(260-508)MMP-19 has the tendency to autoactivate, whereby the Lys(97) Tyr(98) peptide bond is hydrolyzed, resulting in free catalytic domain. Mutation of two residues (Glu(88) --> Pro and Pro(90) --> Val) within the propeptide latency motif did not prevent autoactivation but the autolysis rate was somewhat reduced. Analysis of the substrate specificity revealed that the catalytic domain of MMP-19 was able to hydrolyze the general MMP substrate Mca-Pro-Leu-Gly-Dpa-Ala Arg-NH(2) and, with higher efficiency, the stromelysin substrate Mca-Pro-Leu-Ala Nva-Dpa-Ala-Arg-NH(2). Kinetic analysis of the interactions of the catalytic domain of MMP-19 with the natural MMP inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), showed strong inhibition using TIMP-2, TIMP-3, and TIMP-4, while TIMP-1 was less efficient. We also demonstrated that synthetic hydroxamic acid-based compounds efficiently inhibited the enzyme. The catalytic domain of MMP-19 was able to hydrolyze the basement membrane components type IV collagen, laminin, and nidogen, as well as the large tenascin-C isoform, fibronectin, and type I gelatin in vitro, suggesting that MMP-19 is a potent proteinase capable of hydrolyzing a broad range of extracellular matrix components. Neither the catalytic domain nor the full-length MMP-19 was able to degrade triple-helical collagen. Finally, and in contrast to studies with other MMPs, MMP-19 catalytic domain was not able to activate any of the latent MMPs tested in vitro. PMID- 10809723 TI - Distinct isoforms of the cofactor BAG-1 differentially affect Hsc70 chaperone function. AB - In the mammalian cytosol and nucleus the activity of the molecular chaperone Hsc70 is regulated by chaperone cofactors that modulate ATP binding and hydrolysis by Hsc70. Among such cofactors is the anti-apoptotic protein BAG-1. Remarkably, BAG-1 is expressed as multiple isoforms, which are distinguished by their amino termini. We investigated whether distinct isoforms differ with respect to their Hsc70-regulating activity. By comparing the mainly cytosolic isoforms BAG-1M and BAG-1S, opposite effects of the two isoforms were observed in chaperone-assisted folding reactions. Whereas BAG-1M was found to inhibit the Hsc70-mediated refolding of nonnative polypeptide substrates, the BAG-1S isoform stimulated Hsc70 chaperone activity. The opposite effects are not due to differences in the regulation of the ATPase activity of Hsc70 by the two isoforms. Both isoforms stimulated ATP hydrolysis by Hsc70 in an Hsp40-dependent manner through an acceleration of ADP-ATP exchange. Our results reveal that the different amino termini of the distinct BAG-1 isoforms determine the outcome of an Hsc70-mediated folding event, most likely by transiently interacting with the polypeptide substrate. Employing isoforms of a cofactor with different substrate binding properties appears to provide the means to influence the chaperone function of Hsc70 in addition to modulating its ATPase cycle. PMID- 10809724 TI - PKN binds and phosphorylates human papillomavirus E6 oncoprotein. AB - The high risk human papillomaviruses (HPVs) are associated with carcinomas of cervix and other genital tumors. Previous studies have identified two viral oncoproteins E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high risk HPV E6 protein to immortalize human mammary epithelial cells has provided a single gene model to study the mechanisms of E6 induced oncogenic transformation. In recent years, it has become clear that in addition to E6-induced degradation of p53 tumor suppressor protein, other targets of E6 are required for mammary epithelial cells immortalization. Using the yeast two-hybrid system, we have identified a novel interaction of HPV16 E6 with protein kinase PKN, a fatty acid- and Rho small G protein-activated serine/threonine kinase with a catalytic domain highly homologous to protein kinase C. We demonstrate direct binding of high risk HPV E6 proteins to PKN in wheat-germ lysate in vitro and in 293T cells in vivo. Importantly, E6 proteins of high risk HPVs but not low risk HPVs were able to bind PKN. Furthermore, all the immortalization-competent and many immortalization-non-competent E6 mutants bind PKN. These data suggest that binding to PKN may be required but not sufficient for immortalizing normal mammary epithelial cells. Finally, we show that PKN phosphorylates E6, demonstrating for the first time that HPV E6 is a phosphoprotein. Our finding suggests a novel link between HPV E6 mediated oncogenesis and regulation of a well known phosphorylation cascade. PMID- 10809725 TI - Why does threonine, and not serine, function as the active site nucleophile in proteasomes? AB - Proteasomes belong to the N-terminal nucleophile group of amidases and function through a novel proteolytic mechanism, in which the hydroxyl group of the N terminal threonines is the catalytic nucleophile. However, it is unclear why threonine has been conserved in all proteasomal active sites, because its replacement by a serine in proteasomes from the archaeon Thermoplasma acidophilum (T1S mutant) does not alter the rates of hydrolysis of Suc-LLVY-amc (Seemuller, E., Lupas, A., Stock, D., Lowe, J., Huber, R., and Baumeister, W. (1995) Science 268, 579-582) and other standard peptide amide substrates. However, we found that true peptide bonds in decapeptide libraries were cleaved by the T1S mutant 10 fold slower than by wild type (wt) proteasomes. In degrading proteins, the T1S proteasome was 3.5- to 6-fold slower than the wt, and this difference increased when proteolysis was stimulated using the proteasome-activating nucleotidase (PAN) ATPase complex. With mutant proteasomes, peptide bond cleavage appeared to be rate-limiting in protein breakdown, unlike with wt. Surprisingly, a peptide ester was hydrolyzed by both particles much faster than the corresponding amide, and the T1S mutant cleaved it faster than the wt. Moreover, the T1S mutant was inactivated by the ester inhibitor clasto-lactacystin-beta-lactone severalfold faster than the wt, but reacted with nonester irreversible inhibitors at similar rates. T1A and T1C mutants were completely inactive in all these assays. Thus, proteasomes lack additional active sites, and the N-terminal threonine evolved because it allows more efficient protein breakdown than serine. PMID- 10809726 TI - Microtubule-interfering agents stimulate the transcription of cyclooxygenase-2. Evidence for involvement of ERK1/2 AND p38 mitogen-activated protein kinase pathways. AB - We investigated whether microtubule-interfering agents (MIAs: taxol, colchicine, nocodazole, vinblastine, vincristine, 17-beta-estradiol, 2-methoxyestradiol) altered cyclooxygenase-2 (COX-2) expression in human mammary epithelial cells. MIAs enhanced prostaglandin E(2) synthesis and increased levels of COX-2 protein and mRNA. Nuclear run-off assays revealed increased rates of COX-2 transcription after treatment with MIAs. Calphostin C, an inhibitor of protein kinase C, blocked the induction of COX-2 by MIAs. The stimulation of COX-2 promoter activity by MIAs was inhibited by overexpressing dominant negative forms of Rho and Raf-1. MIAs stimulated ERK, JNK, and p38 mitogen-activated protein kinases (MAPK); pharmacological inhibitors of MAPK kinase and p38 MAPK blocked the induction of COX-2 by MIAs. Overexpressing dominant negative forms of ERK1 or p38 MAPK inhibited MIA-mediated activation of the COX-2 promoter. MIAs stimulated the binding of the activator protein-1 transcription factor complex to the cyclic AMP response element in the COX-2 promoter. A dominant negative form of c-Jun inhibited the activation of the COX-2 promoter by MIAs. Additionally, cytochalasin D, an agent that inhibits actin polymerization, stimulated COX-2 transcription by the same signaling pathway as MIAs. Thus, microtubule- or actin interfering agents stimulated MAPK signaling and activator protein-1 activity. This led, in turn, to induction of COX-2 gene expression via the cyclic AMP response element site in the COX-2 promoter. PMID- 10809727 TI - Eukaryotic selenocysteine incorporation follows a nonprocessive mechanism that competes with translational termination. AB - The synthesis of eukaryotic selenoproteins involves the recoding of an internal UGA codon as a site for selenocysteine incorporation. This recoding event is directed by a selenocysteine insertion sequence in the 3'-untranslated region. Because UGA also functions as a signal for peptidyl-tRNA hydrolysis, we have investigated how the rates of translational termination and selenocysteine incorporation relate to cis-acting elements in the mRNA as well as to trans acting factors in the cytoplasm. We used cis-elements from the phospholipid glutathione peroxidase gene as the basis for this work because of its relatively high efficiency of selenocysteine incorporation. The last two codons preceding the UGA were found to exert a far greater influence on selenocysteine incorporation than nucleotides downstream of it. The efficiency of selenocysteine incorporation was generally much less than 100% but could be partially enhanced by concomitant overexpression of the tRNA(Sec) gene. The combination of two or three UGA codons in one reading frame led to a dramatic reduction in the yield of full-length protein. It is therefore unlikely that multiple incorporations of selenocysteine are processive with respect to the mode of action of the ribosomal complex binding to the UGA site. These observations are discussed in terms of the mechanism of selenoprotein synthesis and its ability to compete with termination at UGA codons. PMID- 10809728 TI - Recombinant laminin-8 (alpha(4)beta(1)gamma(1)). Production, purification,and interactions with integrins. AB - Laminins are a large family of heterotrimeric extracellular matrix glycoproteins that, in addition to having structural roles, take part in the regulation of processes such as cell migration, differentiation, and proliferation. The laminin alpha(4) chain is widely distributed both in adults and during development in tissues such as cardiac, skeletal and smooth muscle fibers, vascular endothelia, lungs, and in peripheral nerves. It can associate with laminin beta(1)/gamma(1) chains to form laminin-8 and with the beta(2)/gamma(1) chains to form laminin-9. Functional studies on these laminins have been hampered by poor availability of the protein in pure and soluble forms. To facilitate studies on laminin-8, recombinant laminin-8 was produced in a mammalian expression system, purified and shown to form native Y-shaped molecules in rotary shadowing electron microscopy. Integrins mediating cell adhesion to laminin-8 were identified using function blocking mAbs. The integrin specificities were found to differ somewhat from that of laminin-1. Integrin alpha(6)beta(1) was found to be a major mediator of adhesion of HT-1080 and cultured capillary endothelial cells to laminin-8. Considering the expression patterns of laminin-8 and integrin alpha(6)beta(1) it is likely that the former is a ligand for the latter in vivo as well. PMID- 10809729 TI - Alteration around the active site of rhodanese during urea-induced denaturation and its implications for folding. AB - The enzyme rhodanese contains two globular domains connected by a tether region and associated by strong hydrophobic interactions. The protein has proven to be very difficult to refold without assistance to prevent oxidation and aggregation. For this study, the active site cysteine 247, near the interdomain region, was modified with the environmentally sensitive fluorescent probe, 2-(4' (iodoacetamido)anilino)naphthalene-6-sulfonic acid (IAANS), to yield a derivative that reversibly unfolds. Structural transitions during urea unfolding/refolding were complex and multiphasic. Increasing urea concentrations increased the IAANS fluorescence intensity and polarization. Both values reached maxima at approximately 4 m urea, where there is a concomitant large exposure of hydrophobic sites as reported by both IAANS and the noncovalent fluorescent probe, bis-ANS. The exposure of the hydrophobic sites arises from the decrease in strong interaction between the domain interfaces, which lead to their partial separation. This correlates with the loss of activity of the unlabeled enzyme. Above 4.5 m urea, there is progressive loss of rigid, hydrophobic surfaces, and both fluorescence and polarization of IAANS decrease, with accompanying loss of secondary structure. These results are consistent with a folding model in which there is an initial, rapid hydrophobic collapse of the denatured form to an intermediate with native like secondary structure, with exposed interdomain, hydrophobic surfaces. This step is followed by adjustment of the domain-domain interactions and the proper positioning of reduced cysteine 247 at the active site. PMID- 10809730 TI - Positive and negative control of multidrug resistance by the Sit4 protein phosphatase in Kluyveromyces lactis. AB - The nuclear gene encoding the Sit4 protein phosphatase was identified in the budding yeast Kluyveromyces lactis. K. lactis cells carrying a disrupted sit4 allele are resistant to oligomycin, antimycin, ketoconazole, and econazole but hypersensitive to paromomycin, sorbic acid, and 4-nitroquinoline-N-oxide (4-NQO). Overexpression of SIT4 leads to an elevation in resistance to paromomycin and to lesser extent tolerance to sorbic acid, but it has no detectable effect on resistance to 4-NQO. These observations suggest that the Sit4 protein phosphatase has a broad role in modulating multidrug resistance in K. lactis. Expression or activity of a membrane transporter specific for paromomycin and the ABC pumps responsible for 4-NQO and sorbic acid would be positively regulated by Sit4p. In contrast, the function of a Pdr5-type transporter responsible for ketoconazole and econazole extrusion, and probably also for efflux of oligomycin and antimycin, is likely to be negatively regulated by the phosphatase. Drug resistance of sit4 mutants was shown to be mediated by ABC transporters as efflux of the anionic fluorescent dye rhodamine 6G, a substrate for the Pdr5-type pump, is markedly increased in sit4 mutants in an energy-dependent and FK506-sensitive manner. PMID- 10809731 TI - The role of the pleckstrin homology domain in membrane targeting and activation of phospholipase Cbeta(1). AB - Current studies involve an investigation of the role of the pleckstrin homology (PH) domain in membrane targeting and activation of phospholipase Cbeta(1) (PLCbeta(1)). Here we report studies on the membrane localization of the isolated PH domain from the amino terminus of PLCbeta(1) (PLCbeta(1)-PH) using fluorescence microscopy of a green fluorescent protein fusion protein. Whereas PLCbeta(1)-PH does not localize to the plasma membrane in serum-starved cells, it undergoes a rapid but transient migration to the plasma membrane upon stimulation of cells with serum or lysophosphatidic acid (LPA). Regulation of the plasma membrane localization of PLCbeta(1)-PH by phosphoinositides was also investigated. PLCbeta(1)-PH was found to bind phosphatidylinositol 3-phosphate most strongly, whereas other phosphoinositides were bound with lower affinity. The plasma membrane localization of PLCbeta(1)-PH induced by serum and LPA was blocked by wortmannin pretreatment and by LY294002. In parallel, activation of PLCbeta by LPA was inhibited by wortmannin, by LY294002, or by the overexpression of PLCbeta(1)-PH. Microinjection of betagamma subunits of G proteins in serum starved cells induced the translocation of PLCbeta(1)-PH to the plasma membrane. These results demonstrate that a cooperative mechanism involving phosphatidylinositol 3-phosphate and the Gbetagamma subunit regulates the plasma membrane localization and activation of PLCbeta(1)-PH. PMID- 10809732 TI - Glycosylphosphatidylinositol-anchored glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall. AB - A novel 1,3-beta-glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus was recently characterized. This enzyme splits internally a 1,3-beta glucan molecule and transfers the newly generated reducing end to the non reducing end of another 1, 3-beta-glucan molecule forming a 1,3-beta linkage, resulting in the elongation of 1,3-beta-glucan chains. The GEL1 gene encoding this enzyme was cloned and sequenced. The predicted amino acid sequence of Gel1p was homologous to several yeast protein families encoded by GAS of Saccharomyces cerevisiae, PHR of Candida albicans, and EPD of Candida maltosa. Although the expression of these genes is required for correct morphogenesis in yeast, the biochemical function of the encoded proteins was unknown. The biochemical assays performed on purified recombinant Gas1p, Phr1p, and Phr2p showed that these proteins have a 1,3-beta-glucanosyltransferase activity similar to that of Gel1p. Biochemical data and sequence analysis have shown that Gel1p is attached to the membrane through a glycosylphosphatidylinositol in a similar manner as the yeast homologous proteins. The activity has been also detected in membrane preparations, showing that this 1,3-beta-glucanosyltransferase is indeed active in vivo. Our results show that transglycosidases anchored to the plasma membrane via glycosylphosphatidylinositols can play an active role in fungal cell wall synthesis. PMID- 10809733 TI - Inactivation of HIV-1 nucleocapsid protein P7 by pyridinioalkanoyl thioesters. Characterization of reaction products and proposed mechanism of action. AB - The synthesis and antiviral properties of pyridinioalkanoyl thioester (PATE) compounds that target nucleocapsid p7 protein (NCp7) of the human immunodeficiency virus type 1 (HIV-1) have been described previously (Turpin, J. A., Song, Y., Inman, J. K., Huang, M., Wallqvist, A., Maynard, A., Covell, D. G., Rice, W. G., and Appella, E. (1999) J. Med. Chem. 42, 67-86). In the present study, fluorescence and electrospray ionization-mass spectrometry were employed to determine the mechanism of modification of NCp7 by two lead compounds, N-[2-(5 pyridiniovaleroylthio)benzoyl]sulfacetamide bromide and N-[2-(5 pyridiniovaleroylthio)benzoyl]-4-(4-nitrophenylsulfonyl )anili ne bromide (compounds 45 and 47, respectively). Although both compounds exhibit antiviral activity in cell-based assays, we failed to detect appreciable ejection of zinc from NCp7 under conditions in which previously described NCp7-active disulfides readily eject zinc. However, upon "activation" by Ag(+), compound 45 reacted with NCp7 resulting in the zinc ejection from both zinc fingers. The reaction followed a two-step mechanism in which zinc was ejected from the carboxyl-terminal zinc finger faster than from the amino-terminal zinc finger. Both compounds covalently modified the protein with pyridinioalkanoyl groups. Compound 45 modified cysteines 36 and 49 of the carboxyl-terminal zinc finger. The results obtained herein demonstrate that PATE compounds can be constructed that selectively target only one of the two zinc fingers of NCp7, thus providing an impetus to pursue development of highly selective zinc finger inhibitors. PMID- 10809734 TI - Cloning and characterization of COX18, a Saccharomyces cerevisiae PET gene required for the assembly of cytochrome oxidase. AB - Nuclear mutants of Saccharomyces cerevisiae assigned to complementation group G34 are respiratory-deficient and lack cytochrome oxidase activity and the characteristic spectral peaks of cytochromes a and a(3). The corresponding gene was cloned by complementation, sequenced, and identified as reading frame YGR062C on chromosome VII. This gene was named COX18. The COX18 gene product is a polypeptide of 316 amino acids with a putative amino-terminal mitochondrial targeting sequence and predicted transmembrane domains. Respiratory chain carriers other than cytochromes a and a(3) and the ATPase complex are present at near wild-type levels in cox18 mutants, indicating that the mutations specifically affect cytochrome oxidase. The synthesis of Cox1p and Cox3p in mutant mitochondria is normal whereas Cox2p is barely detected among labeled mitochondrial polypeptides. Transcription of COX2 does not require COX18 function, and a chimeric COX3-COX2 mRNA did not suppress the respiratory defect in the null mutant, indicating that the mutation does not impair transcription or translation of the mRNA. Western analysis of cytochrome oxidase subunits shows that inactivation of the COX18 gene greatly reduces the steady state amounts of subunit 2 and results in variable decreases in other subunits of cytochrome oxidase. A gene fusion expressing a biotinylated form of Cox18p complements cox18 mutants. Biotinylated Cox18p is a mitochondrial integral membrane protein. These results indicate Cox18p to be a new member of a group of mitochondrial proteins that function at a late stage of the cytochrome oxidase assembly pathway. PMID- 10809735 TI - Involvement of the acute phase protein alpha 1-acid glycoprotein in nonspecific resistance to a lethal gram-negative infection. AB - Resistance to gram-negative infection can be induced by pretreating animals with several agents such as turpentine and interleukin (IL)-1. Because these agents are powerful inducers of acute phase proteins, we wondered whether these proteins, more particularly alpha(1)-acid glycoprotein (alpha(1)-AGP), are involved in nonspecific resistance to infection. Turpentine and IL-1 protect completely against a lethal challenge of Klebsiella pneumoniae when given 48 and 12-48 h before the challenge, respectively. alpha(1)-AGP induction in the serum reached peak values 48 h after turpentine and 12-48 h after IL-1 injection. Administration of alpha(1)-AGP, 2 h before a challenge of K. pneumoniae, significantly increased the survival. Numbers of bacteria cultured from blood and organs were significantly lower in mice pretreated with a protective dose of turpentine, IL-1, or alpha(1)-AGP. These data suggest that alpha(1)-AGP is a possible mediator in turpentine- or IL-1-induced protection because time points of maximal induction of alpha(1)-AGP by turpentine or IL-1 and of optimal protection by alpha(1)-AGP coincide. Transgenic overexpression of rat alpha(1) AGP protected mice from a K. pneumoniae infection. Bacterial counts in blood and organs were significantly lower in transgenic mice, and only in control mice were large necrotic areas, apoptosis, and blood clots observed in the spleen. Our data suggest that alpha(1)-AGP prevents gram-negative infections and may be an essential component in nonspecific resistance to infection. PMID- 10809736 TI - Dissection of protein linkage between keratins and pinin, a protein with dual location at desmosome-intermediate filament complex and in the nucleus. AB - Pinin is a cell adhesion-associated and nuclear protein that has been shown to localize in the vicinity of intermediate filament (IF) convergence upon the cytoplasmic face of the desmosomal plaque as well as in the nucleus. The localization of pinin to the desmosomes has been correlated with the reinforcement of intercellular adhesion and increased IF organization. In this study, keratins 18, 8, and 19 were identified to interact with the amino end domain of pinin in a two-hybrid screening. Further truncation analyses indicated that the 2B domain of keratin contains the sequence responsible for interacting with pinin. The amino end of pinin (residues 1-98) is sufficient to bind to keratin. Point mutation analyses revealed two essential residues within the pinin fragment 1-98, leucine 8 and leucine 19, for the interaction with keratin. Finally, in vitro protein overlay binding assays confirmed the direct interaction of the amino end domain of pinin with keratins, while pinin mutant L8P GST fusion protein failed to bind to keratins in the overlay assay. Coupled with our previous morphological observations and transfection studies, these data suggest that pinin may play a role in epithelial cell adhesion and the IF complex through a direct interaction with the keratin filaments. PMID- 10809737 TI - The sensitivity of RNA polymerase II in elongation complexes to C-terminal domain phosphatase. AB - The phosphorylation state of the carboxyl-terminal domain (CTD) of the largest RNA polymerase (RNAP) II subunit plays an important role in the regulation of transcript elongation. This report examines the sensitivity of RNAP II to dephosphorylation by CTD phosphatase (CTDP) and addresses factors that regulate its sensitivity. The CTDP sensitivity of RNAP IIO in paused elongation complexes on a dC-tailed template does not significantly differ from that of free RNAP IIO. RNAP IIO contained in elongation complexes that initiate transcription from the adenovirus-2 major late promoter in the presence of a nuclear extract is relatively resistant to dephosphorylation. Complexes treated with 1% Sarkosyl remain elongation-competent but demonstrate a 5-fold increase in CTDP sensitivity. Furthermore, the sensitivity of RNAP IIO in both control and Sarkosyl-treated elongation complexes is dependent on their position relative to the start site of transcription. Elongation complexes 11-24 nucleotides downstream are more sensitive to dephosphorylation than complexes 50-150 nucleotides downstream. The incubation of Sarkosyl-treated elongation complexes with nuclear extract restores the original resistance to dephosphorylation. These results suggest that a conformational change occurs in RNAP II as it clears the promoter, which results in an increased resistance to dephosphorylation. Furthermore, the sensitivity to dephosphorylation can be modulated by a factor(s) present in the nuclear extract. PMID- 10809738 TI - The role of ERp57 in disulfide bond formation during the assembly of major histocompatibility complex class I in a synchronized semipermeabilized cell translation system. AB - We have established a semipermeabilized cell system that reproduces the folding and assembly of a major histocompatibility complex (MHC) class I complex as it would occur in the intact cell. The translation of the MHC class I heavy chain (HLA-B27) in this system was synchronized allowing the folding and assembly of polypeptide chains synthesized within a short time frame to be analyzed. This has enabled us to dissect the time course of interaction of both disulfide and nondisulfide-bonded heavy chain with various molecular chaperones during its assembly in a functionally intact endoplasmic reticulum. The results demonstrate that unassembled, nondisulfide-bonded forms of heavy chain interact initially with calnexin. A later and more prolonged interaction of calreticulin, specifically with assembled, disulfide-bonded heavy chain, highlights distinct differences in the roles of these two proteins in the assembly of MHC class I molecules. We also demonstrate that the thiol-dependent reductase ERp57 initially interacts with nondisulfide-bonded heavy chain, but this rapidly becomes disulfide-bonded and indicates that heavy chain folding occurs during its interaction with ERp57. In addition, we also confirm a direct interaction between MHC class I heavy chain and tapasin, emphasizing the role that this protein plays in the later stages of MHC class I assembly. PMID- 10809739 TI - Hyaluronan anchoring and regulation on the surface of vascular endothelial cells is mediated through the functionally active form of CD44. AB - CD44 on lymphocytes binding to its carbohydrate ligand hyaluronan can mediate primary adhesion (rolling interactions) of lymphocytes on vascular endothelial cells. This adhesion pathway is utilized in the extravasation of activated T cells from the blood into sites of inflammation and therefore influences patterns of lymphocyte homing and inflammation. Hyaluronan is a glycosaminoglycan found in the extracellular matrix and is involved in a number of biological processes. We have shown that the expression of hyaluronan on the surface of endothelial cells is inducible by proinflammatory cytokines. However, the manner through which hyaluronan is anchored to the endothelial cell surface so that it can resist shear forces and the mechanism of the regulation of the level of hyaluronan on the cell surface has not been investigated. In order to characterize potential hyaluronan receptors on endothelial cells, we performed analyses of cell surface staining by flow cytometry on intact endothelial cells and ligand blotting assays using membrane fractions. Hyaluronan binding activity was detected as a major species corresponding to the size of CD44, and this was confirmed to be the same by Western blotting and immunoprecipitation. Moreover, alterations in the surface level of hyaluronan after tumor necrosis factor-alpha stimulation is regulated primarily by changes in the cell surface levels of the hyaluronan-binding form of CD44. In laminar flow assays, lymphoid cells specifically roll on hyaluronan anchored by purified CD44 coated on glass tubes, indicating that the avidity of the endothelial CD44/hyaluronan interaction is sufficient to support rolling adhesions under conditions mimicking physiologic shear forces. Together these studies show that CD44 serves to anchor hyaluronan on endothelial cell surfaces, that activation of CD44 is a major regulator of endothelial surface hyaluronan expression, and that the non-covalent interaction between CD44 and hyaluronan is sufficient to provide resistance to shear under physiologic conditions and thereby support the initial steps of lymphocyte extravasation. PMID- 10809740 TI - Modulation of Rho and cytoskeletal protein attachment to membranes by a prenylcysteine analog. AB - The GTPases Rho regulate the assembly of polymerized actin structures. Their C terminal sequences end with the CAAX motif that undergo a lipidation of the cysteine residue. Analogs to the C-terminal ends of Rho proteins, N-acetyl-S-all trans, trans-farnesyl-L-cysteine and N-acetyl-S-all-trans-geranylgeranyl-L cysteine, wereused to analyze the role of prenylation in their membrane association. Silver-stained gels indicated that N-acetyl-S-all-trans geranylgeranyl-L-cysteine treatment released only a few proteins of 20, 46, and 60 kDa. Western blot analysis showed that N-acetyl-S-all-trans-geranylgeranyl-L cysteine released RhoB (10%), RhoA (28%), and Cdc42 (95%) from membranes, whereas N-acetyl-S-all-trans and trans-farnesyl-L-cysteine did not. Rab1, which possesses two geranylgeranyl groups, was also strongly extracted by N-acetyl-S-all-trans geranylgeranyl-L-cysteine, whereas Ras, which is farnesylated, was not. Furthermore, N-acetyl-S-all-trans-geranylgeranyl-L-cysteine was very efficient (95%) in dissociating actin and tubulin from membranes but not integral membrane protein P-glycoprotein and sodium/phosphate cotransporter NaP(i)-2. The extraction of Rho and cytoskeletal proteins occurred below the critical micellar concentration of N-acetyl-S-all-trans-geranylgeranyl-L-cysteine. Membrane treatments with 0.7 m KI totally extracted actin, whereas 70% of Cdc42 was released. Actin was, however, insoluble in Triton X-100-treated membranes, whereas this detergent extracted (80%) Cdc42. These data show that Rho proteins and actin are not physically bound together and suggest that their extraction from membranes by N-acetyl-S-all-trans-geranylgeranyl-L-cysteine likely occurs via different mechanisms. PMID- 10809741 TI - S-adenosylmethionine and Pneumocystis carinii. AB - We previously reported that S-adenosylmethionine (AdoMet), a key molecule in methylation reactions and polyamine biosynthesis, enhances axenic culture of the AIDS-associated opportunistic fungal pathogen Pneumocystis carinii. Here we report that AdoMet is absolutely required for continuous growth. Two transporters are present, one high affinity, K(m) = 4.5 microm, and one low affinity, K(m) = 333 microm. The physiologically relevant high affinity transporter has a pH optimum of 7.5 and no related natural compounds compete for uptake. Transport is 98% inhibited at 4 degrees C, 24% inhibited by 20 mm sodium azide, and 95% inhibited by the combination of 20 mm sodium azide and 1 mm salicylhydroxamic acid; thus transport is active and dependent on both a cytochrome chain and an alternative oxidase. In vitro, AdoMet is used at a rate of 1. 40 x 10(7) molecules cell(-1) min(-1). AdoMet synthetase activity was not detected by a sensitive radiolabel incorporation assay capable of detecting 0.1% of the activity in rat liver. In addition, the AdoMet plasma concentration of rats is inversely correlated with the number of P. carinii in the lungs. These findings demonstrate that P. carinii is an AdoMet auxotroph. The uptake and metabolism of this compound are rational chemotherapeutic targets. PMID- 10809742 TI - Peptides selected to bind the Gal80 repressor are potent transcriptional activation domains in yeast. AB - The activation domain of the yeast Gal4 protein binds specifically to the Gal80 repressor and is also thought to associate with one or more coactivators in the RNA polymerase II holoenzyme and chromatin remodeling machines. This is a specific example of a common situation in biochemistry where a single protein domain can interact with multiple partners. Are these different interactions related chemically? To probe this point, phage display was employed to isolate peptides from a library based solely on their ability to bind Gal80 protein in vitro. Peptide-Gal80 protein association is shown to be highly specific and of moderate affinity. The Gal80 protein-binding peptides compete with the native activation domain for the repressor, suggesting that they bind to the same site. It was then asked if these peptides could function as activation domains in yeast when tethered to a DNA binding domain. Indeed, this is the case. Furthermore, one of the Gal80-binding peptides binds directly to a domain of the Gal11 protein, a known coactivator. The fact that Gal80-binding peptides are functional activation domains argues that repressor binding and activation/coactivator binding are intimately related properties. This peptide library-based approach should be generally useful for probing the chemical relationship of different binding interactions or functions of a given native domain. PMID- 10809743 TI - The transgenic expression of highly inhibitory monomeric forms of phospholamban in mouse heart impairs cardiac contractility. AB - Transgenic mice were generated with cardiac-specific overexpression of the monomeric, dominant-acting, superinhibitory L37A and I40A mutant forms of phospholamban (PLN), and their phenotypes were compared with wild-type (wt) mice or 2-fold overexpressors of wt PLN (wtOE). The level of PLN monomer in cardiac microsomes was increased 11-13-fold, and the apparent affinity of the sarco(endo)plasmic reticulum Ca(2+)-ATPase for Ca(2+) was decreased from pCa 6.22 in wt or 6.12 in wtOE to 5.81 in L37A and 5.72 in I40A. Basal physiological parameters, measured in isolated myocytes, indicated a significant reduction in the rates of shortening (+dL/dt) and relengthening (-dL/dt). Hemodynamic measurements indicated that peak systolic pressure was unaffected but that pressure changes (+dP/dt and -dP/dt) were lowered significantly in both mutant lines, and relaxation time (tau) was also lengthened significantly. Echocardiography for both mutants showed depressed systolic function and an increase in left ventricular mass of over 1.4-fold. Significant decreases in left ventricular shortening fraction and velocity of circumferential shortening and increases in ejection time were corrected by isoproterenol. The use of antibodies specific against Ser(16)- and Thr(17)-PLN peptides showed that phosphorylation of both pentameric and monomeric PLN were increased between 1.2- and 2.4-fold in both the L37A and I40A lines but not in the wtOE line. These observations show that overexpression of superinhibitory mutant forms of PLN causes depression of contractile parameters with induction of cardiac hypertrophy, as assessed with echocardiography. PMID- 10809744 TI - Reduced ethanol inhibition of N-methyl-D-aspartate receptors by deletion of the NR1 C0 domain or overexpression of alpha-actinin-2 proteins. AB - The depressant actions of ethanol on central nervous system activity appear to be mediated by its actions on a number of important membrane associated ion channels including the N-methyl-d-aspartate (NMDA) subtype of ionotropic glutamate receptor. Although no specific site of action for ethanol on the NMDA receptor has been found, previous studies suggest that the ethanol sensitivity of the receptor may be affected by intracellular C-terminal domains of the receptor that regulate the calcium-dependent inactivation of the receptor. In the present study, co-expression of the NR2A subunit and an NR1 subunit that lacks the alternatively spliced intracellular C1 cassette did not reduce the effects of ethanol on channel function as measured by patch-clamp electrophysiology. Full inhibition was also observed in cells expressing an NR1 subunit truncated at the end of the C0 domain (NR1(863stop)). However, the inhibitory effects of ethanol were reduced by expression of an NR1 C0 domain deletion mutant (NR1(Delta839 863)), truncation mutant (NR1(858stop)), or a triple-point mutant (Arg to Ala, Lys to Ala, and Asn to Ala at 859-861) previously shown to significantly reduce calcium-dependent inactivation. A similar reduction in the effects of ethanol on wild-type NR1/2A but not NR1/2B or NR1/2C receptors was observed after co expression of full-length or truncated human skeletal muscle alpha-actinin-2 proteins that produce a functional knockout of the C0 domain. The effects of ethanol on hippocampal and cortical NMDA-induced currents were similarly attenuated in low calcium recording conditions, suggesting that a C0 domain dependent process may confer additional ethanol sensitivity to NMDA receptors. PMID- 10809745 TI - Physical interactions between phospholamban and sarco(endo)plasmic reticulum Ca2+ ATPases are dissociated by elevated Ca2+, but not by phospholamban phosphorylation, vanadate, or thapsigargin, and are enhanced by ATP. AB - Previous co-immunoprecipitation studies (Asahi, M., Kimura, Y., Kurzydlowski, K., Tada, M., and MacLennan, D. H. (1999) J. Biol. Chem. 274, 32855-32862) revealed that physical interactions between phospholamban (PLN) and the fast-twitch skeletal muscle sarco(endo)plasmic reticulum Ca(2+) ATPase (SERCA1a) were retained, even with PLN monoclonal antibody 1D11 bound to an epitope lying between PLN residues 7 and 17. Because the 1D11 antibody relieves inhibitory interaction between the two proteins, it was of interest to determine whether PLN phosphorylation or elevation of Ca(2+), which also relieves inhibitory interactions between PLN and SERCA, would disrupt physical interactions. Co immunoprecipitation was measured in the presence of increasing concentrations of Ca(2+) or after phosphorylation of PLN by protein kinase A. Physical interactions were dissociated by elevated Ca(2+) but not by PLN phosphorylation. The addition of ATP enhanced interactions between PLN and SERCA. The further addition of vanadate and thapsigargin, both of which stabilize the E(2) conformation, did not diminish binding of PLN to SERCA. These data suggest that physical interactions between PLN and SERCA are stable when SERCA is in the Ca(2+)-free E(2) conformation but not when it is in the E(1) conformation and that phosphorylation of PLN does not dissociate physical interactions between PLN and SERCA. PMID- 10809746 TI - Coactivator-vitamin D receptor interactions mediate inhibition of the atrial natriuretic peptide promoter. AB - We have discovered a role for coactivators binding to the AF-2 surface of the vitamin D receptor (VDR) in its negative effects on gene transcription. We tested nine amino acid residues (Ser(235), Ile(242), Lys(246), Asp(253), Ile(260), Leu(263), Leu(417), Leu(419), and Glu(420)) in human VDR which, based on homology to the human thyroid hormone receptor, would be predicted to lie in or near the coactivator-binding site. Mutation of six of these residues in VDR resulted in loss of both the activation (assessed with a transfected DR3 TK luciferase reporter) and inhibition (assessed with an hANPCAT reporter) functions of the receptor when tested in cultured neonatal rat atrial myocytes and HeLa cells. Collectively, these mutations also suppressed association of VDR with the coactivators GRIP1 and steroid receptor coactivator 1 in vitro but had little or no effect on ligand binding, heterodimerization with the retinoid X receptor, or association with a VDR-specific DNA recognition element. Co-transfection with GRIP1 or steroid receptor coactivator 1 amplified both the positive and negative responses to wild type VDR but had little or no effect on the functionally impaired mutants described above. The interaction between VDR and GRIP1 proved to be heavily dependent upon the integrity of nuclear box III in the latter protein. Mutations in this region of GRIP1 impaired its ability to associate with VDR in vitro and to amplify VDR activity in intact cells. These studies establish a role for coactivators recruited to the same receptor surface in both the activating and inhibitory activity of the liganded receptor. PMID- 10809747 TI - Probing the active site of the hepatitis C virus serine protease by fluorescence resonance energy transfer. AB - A serine protease domain contained within the viral NS3 protein is a key player in the maturational processing of the hepatitis C virus polyprotein and a prime target for the development of antiviral drugs. In the present work, we describe a dansylated hexapeptide inhibitor of this enzyme. Active site occupancy by this compound could be monitored following fluorescence resonance energy transfer between the dansyl fluorophore and protein tryptophan residues and could be used to 1) unambiguously assess active site binding of NS3 protease inhibitors, 2) directly determine equilibrium and pre-steady-state parameters of enzyme inhibitor complex formation, and 3) dissect, using site-directed mutagenesis, the contribution of single residues of NS3 to inhibitor binding in direct binding assays. The assay was also used to characterize the inhibition of the NS3 protease by its cleavage products. We show that enzyme-product inhibitor complex formation depends on the presence of an NS4A cofactor peptide. Equilibrium and pre-steady-state data support an ordered mechanism of ternary (enzyme-inhibitor cofactor) complex formation, requiring cofactor complexation prior to inhibitor binding. PMID- 10809748 TI - Inhibition of NF-kappaB activity and enhancement of apoptosis by the neuropeptide calcitonin gene-related peptide. AB - Calcitonin gene-related peptide (CGRP) is a neuropeptide produced by the central and peripheral nervous systems and by endocrine cells. CGRP exerts diverse biological effects on the cardiovascular, gastrointestinal, respiratory, central nervous and immune systems. Little is known, however, about the molecular mechanisms that mediate CGRP effects. Using the NFkappaB-luciferase reporter transgenic mice, here we show that CGRP selectively inhibits NF-kappaB-mediated transcription in thymocytes in vitro and in vivo. In contrast, CGRP does not affect transcription mediated by the AP-1 and NFAT transcription factors. CGRP inhibits the accumulation of NF-kappaB complexes in the nucleus by preventing phosphorylation and degradation of the NF-kappaB inhibitor IkappaB. Inhibition of NF-kappaB activity is associated with the induction of apoptosis by CGRP in thymocytes. Together these results demonstrate for the first time the selective implication of the NF-kappaB signaling pathway in the regulatory function of the neuropeptide CGRP. Our study suggests a potential molecular mechanism by which CGRP can induce cell death in thymocytes. PMID- 10809749 TI - A single intracellular cysteine residue is responsible for the activation of the olfactory cyclic nucleotide-gated channel by NO. AB - The activation of cyclic nucleotide-gated (CNG) channels is the final step in olfactory and visual transduction. Previously we have shown that, in addition to their activation by cyclic nucleotides, nitric oxide (NO)-generating compounds can directly open olfactory CNG channels through a redox reaction that results in the S-nitrosylation of a free SH group on a cysteine residue. To identify the target site(s) of NO, we have now mutated the four candidate intracellular cysteine residues Cys-460, Cys-484, Cys-520, and Cys-552 of the rat olfactory rCNG2 (alpha) channel into serine residues. All mutant channels continue to be activated by cyclic nucleotides, but only one of them, the C460S mutant channel, exhibited a total loss of NO sensitivity. This result was further supported by a similar lack of NO sensitivity that we found for a natural mutant of this precise cysteine residue, the Drosophila melanogaster CNG channel. Cys-460 is located in the C-linker region of the channel known to be important in channel gating. Kinetic analyses suggested that at least two of these Cys-460 residues on different channel subunits were involved in the activation by NO. Our results show that one single cysteine residue is responsible for NO sensitivity but that several channel subunits need to be activated for channel opening by NO. PMID- 10809750 TI - Kinetics of smooth muscle heavy meromyosin with one thiophosphorylated head. AB - Actin-activated MgATPase of smooth muscle heavy meromyosin is activated by thiophosphorylation of two regulatory light chains, one on each head domain. To understand cooperativity between heads, we examined the kinetics of heavy meromyosin (HMM) with one thiophosphorylated head. Proteolytic gizzard heavy meromyosin regulatory light chains were partially exchanged with recombinant thiophosphorylated His-tagged light chains, and HMM with one thiophosphorylated head was isolated by nickel-affinity chromatography. In vitro motility was observed. By steady-state kinetic analysis, one-head thiophosphorylated heavy meromyosin had a similar K(m) value for actin but a V(max) value of approximately 50% of the fully thiophosphorylated molecule. However, single turnover analysis, which is not sensitive to small amounts of active heads, showed that one-head thiophosphorylated heavy meromyosin was 46-120 times more active than unphosphorylated HMM but only 7-19% as active as the fully thiophosphorylated molecule. Discrepancy between the single turnover and steady-state values could be explained by a small fraction of rigor heads. These rigor heads would have a large effect on the steady-state kinetics of one-head thiophosphorylated HMM. In summary, thiophosphorylation of one head leads to a molecule with unique intermediate kinetics suggesting that thiophosphorylation of one head cooperatively alters the kinetics of the partner head and vice versa. PMID- 10809751 TI - A mammalian iron ATPase induced by iron. AB - While molecular mechanisms for iron entry and storage within cells have been elucidated, no system to mediate iron efflux has been heretofore identified. We now describe an ATP requiring iron transporter in mammalian cells. (55)Fe is transported into microsomal vesicles in a Mg-ATP-dependent fashion. The transporter is specific for ferrous iron, is temperature- and time-dependent, and detected only with hydrolyzable nucleotides. It differs from all known ATPases and appears to be a P-type ATPase. The Fe-ATPase is localized together with heme oxygenase-1 to microsomal membranes with both proteins greatly enriched in the spleen. Iron treatment markedly induces ATP-dependent iron transport in RAW 264.7 macrophage cells with an initial phase that is resistant to cycloheximide and actinomycin D and a later phase that is inhibited by these agents. Iron release, elicited in intact rats by glycerol-induced rhabdomyolysis, induces ATP-dependent iron transport in the kidney. Mice with genomic deletion of heme oxygenase-1 have selective tissue iron accumulation and display augmented ATP-dependent iron transport in those tissues that accumulate iron. PMID- 10809752 TI - Reconstitution of membranes simulating "glycosignaling domain" and their susceptibility to lyso-GM3. AB - GM3 ganglioside at the surface of mouse melanoma B16 cells is clustered and organized with signal transducer molecules c-Src, Rho A, and focal adhesion kinase (FAK) to form a membrane unit separable from caveolae, which are enriched in cholesterol and caveolin but do not contain GM3 or the above three signal transducers. The GM3-enriched membrane units are involved in GM3-dependent cell adhesion coupled with activation of c-Src, Rho A, and FAK and are termed the "glycosphingolipid signaling domain" or the "glycosignaling domain" (GSD). In order to assess the essential components that display GSD function, membranes with properties similar to those of GSD were reconstituted using GM3, sphingomyelin, and c-Src, with or without other lipid components. The reconstituted membrane thus prepared displayed GM3-dependent adhesion to plates coated with Gg3 or anti-GM3 antibody, resulting in enhanced c-Src phosphorylation (c-Src phosphorylation response). This response in reconstituted membrane depends on GM3 concentration and was not observed when GM3 was absent or replaced with other gangliosides GM1 or GD1a, or with LacCer. The GM3-dependent c-Src phosphorylation response was enhanced when cholesterol and phosphatidylcholine were added. Although GM3, sphingomyelin, and c-Src are essential for GSD function, a small quantity of cholesterol and phosphatidylcholine may act as an auxiliary factor to stabilize membrane. GSD function in terms of GM3-dependent adhesion and signaling was blocked in the presence of lyso-GM3 or its analogue but not psychosine, lactosyl-sphingosine, or lyso-phosphatidylcholine. Such susceptibility of reconstituted GSD to lyso-GM3 and other lyso compounds is the same as GSD of original B16 cells. Thus, functional organization of the reconstituted membrane closely simulates that of GSD in B16 cells, which is based on clustered GM3 organized with c-Src as the essential components. PMID- 10809753 TI - Analysis of a gene encoding Rpn10 of the fission yeast proteasome reveals that the polyubiquitin-binding site of this subunit is essential when Rpn12/Mts3 activity is compromised. AB - Substrates are targeted for proteolysis by the ubiquitin pathway by the addition of a polyubiquitin chain before being degraded by the 26 S proteasome. Previously, a subunit of the proteasome, S5a, was identified that was able to bind to polyubiquitin in vitro and thus proposed to act as a substrate recognition component. Deletion of the corresponding Saccharomyces cerevisiae gene, MCB1/RPN10, rendered cells viable indicating that other proteasomal polyubiquitin receptors must exist. In this study, we describe pus1(+), the fission yeast homologue of RPN10. This gene is also not required for cell viability; however, the Deltapus1 mutant is synthetically lethal with mutations in other proteasomal component-encoding genes, namely mts3, pad1, and mts4 (RPN12, RPN11, and RPN1). Overexpression of pus1(+) is able to rescue mts3-1 at 32 degrees C but overexpression of a cDNA encoding a version of Pus1 that does not bind to polyubiquitin cannot and leads to greatly reduced viability when used to rescue the mts3-1Deltapus1 double mutant. The Mts3 protein was unable to bind to polyubiquitin in vitro, but the Pus1 and Mts3 proteins were found to bind to one another in vitro, which taken together with the genetic data suggests that they are also closely associated in vivo. PMID- 10809754 TI - Inducible NF-kappaB activation is permitted by simultaneous degradation of nuclear IkappaBalpha. AB - Signal-induced phosphorylation and ubiquitination of IkappaBalpha targets this inhibitor of NF-kappaB for proteasome-mediated degradation, thus permitting the release of active NF-kappaB. Upon cell stimulation, NF-kappaB activation results in neotranscription and neosynthesis of its own inhibitor, IkappaBalpha. As reported earlier, the neosynthesized inhibitor is then accumulated in the nucleus, where it rapidly binds to and terminates the function of nuclear NF kappaB upon withdrawal of the stimulus. The present work was aimed at understanding how NF-kappaB activity is preserved while stimuli persist, despite intense, simultaneous IkappaBalpha neosynthesis, which would be expected to end NF-kappaB activity. We here show that incoming IkappaBalpha in the nucleus represents a target for resident nuclear proteasome complexes. Signal-induced, proteasome-dependent degradation of phosphorylated and ubiquitinated IkappaBalpha occurs in the nucleus, thus permitting the onset and persistence of NF-kappaB activity as long as stimulation is maintained. Our results suggest that intranuclear proteolysis of IkappaBalpha is necessarily required to avoid self termination of NF-kappaB activity during cell activation. PMID- 10809755 TI - Morphologic differentiation of HL-60 cells is associated with appearance of RPTPbeta and induction of Helicobacter pylori VacA sensitivity. AB - Phorbol 12-myristate 13-acetate (PMA) induces differentiation of human leukemic HL-60 cells into cells with macrophage-like characteristics and enhances the susceptibility of HL-60 cells to the Helicobacter pylori VacA toxin (de Bernard, M., Moschioni., M., Papini, E., Telford, J. L., Rappuoli, R., and Montecucco, C. (1998) FEBS Lett. 436, 218-222). We examined the mechanism by which HL-60 cells acquire sensitivity to VacA, in particular, looking for expression of RPTPbeta, a VacA-binding protein postulated to be the VacA receptor (Yahiro, K., Niidome, T., Kimura, M., Hatakeyama, T., Aoyagi, H., Kurazono, H., Imagawa, K., Wada, A., Moss, J., and Hirayama, T. (1999) J. Biol. Chem. 274, 36693-36699). PMA induced expression of RPTPbeta mRNA and protein as determined by RNase protection assay and indirect immunofluorescence studies, respectively. Vitamin D(3) and interferon-gamma, which stimulate differentiation of HL-60 cells into monocyte like cells, also induced VacA sensitivity and expression of RPTPbeta mRNA, whereas 1. 2% Me(2)SO and retinoic acid, which stimulated the maturation of HL-60 into granulocyte-like cells, did not. RPTPbeta antisense oligonucleotide inhibited induction of VacA sensitivity and expression of RPTPbeta. Double immunostaining studies also indicated that newly expressed RPTPbeta colocalized with VacA in PMA-treated HL-60 cells. In agreement with these data, BHK-21 cells, which are insensitive to VacA, when transfected with the RPTPbeta cDNA, acquired VacA sensitivity. All data are consistent with the conclusion that acquisition of VacA sensitivity by PMA-treated HL-60 cells results from induction of RPTPbeta, a protein that functions as the VacA receptor. PMID- 10809756 TI - Interactions between the exocytic and endocytic pathways in polarized Madin-Darby canine kidney cells. AB - The compartments involved in polarized exocytosis of membrane proteins are not well defined. In this study we hypothesized that newly synthesized polymeric immunoglobulin receptors are targeted from the trans-Golgi network to endosomes prior to their appearance on the basolateral cell surface of polarized Madin Darby canine kidney cells. To examine this hypothesis, we have used an assay designed to measure the meeting of newly synthesized receptors with a selective population of apical or basolateral endosomes loaded with horseradish peroxidase. We found that in the course of basolateral exocytosis, the wild-type polymeric immunoglobulin receptor is targeted from the trans-Golgi network to apical and basolateral endosomes. Phosphorylation of a Ser residue in the cytoplasmic tail of the receptor is implicated in this process. The biosynthetic pathway of apically sorted polymeric immunoglobulin receptor mutants similarly traversed apical endosomes, raising the possibility that apical receptors are segregated from basolateral receptors in apical endosomes. The post-endocytic pathway of transcytosing and recycling receptors also passed through apical endosomes. Together, these observations are consistent with the possibility that the biosynthetic and endocytic routes merge into endosomes and justify a model suggesting that endosomal recycling processes govern polarized trafficking of proteins traveling in both pathways. PMID- 10809757 TI - Tumor necrosis factor alpha suppresses the induction of connective tissue growth factor by transforming growth factor-beta in normal and scleroderma fibroblasts. AB - Connective tissue growth factor (CTGF) is overexpressed in a variety of fibrotic disorders, presumably secondary to the activation and production of transforming growth factor-beta (TGF-beta), a key inducer of fibroblast proliferation and matrix synthesis. The CTGF gene promoter has a TGF-beta response element that regulates its expression in fibroblasts but not epithelial cells or lymphocytes. Recent studies have shown that the macrophage-produced cytokine tumor necrosis factor alpha (TNFalpha) is necessary to promote inflammation and to induce genes, such as matrix metalloproteinases, involved with the early stages of wound healing. In this study, we examined the ability of TNFalpha to modulate CTGF gene expression. TNFalpha was found to suppress the TGF-beta-induced expression of CTGF protein in cultured normal fibroblasts. The activity of TNFalpha was blocked by NF-kappaB inhibitors. We showed that sequences between -244 and -166 of the CTGF promoter were necessary for both TGF-beta and TNFalpha to modulate CTGF expression. There was a constitutive expression of CTGF by scleroderma fibroblasts that was increased by TGF-beta treatment. Although TNFalpha was able to repress TGF-beta-induced CTGF and collagen synthesis both in normal and scleroderma skin fibroblasts, fibroblasts cultured from scleroderma patients were more resistant to TNFalpha as TNFalpha was unable to suppress the basal level of CTGF expression in scleroderma fibroblasts. Thus, we suspect that the high level of constitutive CTGF expression in scleroderma fibroblasts and its inability to respond to negative regulatory cytokines may contribute to the excessive scarring of skin and internal organs in patients with scleroderma. PMID- 10809758 TI - Induction of neuronal differentiation by p73 in a neuroblastoma cell line. AB - The p53-related p73 and p63 genes encode proteins that share considerable structural and functional homology with p53. Despite similarities, their deletion in mice has different outcomes, implying that the three genes may play distinct roles in vivo. Here we show that endogenous p73 levels increase in neuroblastoma cells induced to differentiate by retinoic acid and that exogenously expressed p73, but not p53, is sufficient to induce both morphological (neurite outgrowth) and biochemical (expression of neurofilaments and neural cell adhesion molecule (N-CAM); down-regulation of N-MYC and up-regulation of pRB) markers of neuronal differentiation. This activity is shared, to different extents, by all p73 isoforms, whereas the transcriptionally inactive mutants of p73 isoforms are ineffective. Conversely, blockage of endogenous p73 isoforms with a dominant negative p73 results in the abrogation of retinoid-induced N-CAM promoter-driven transcription. Our results indicate that the p73 isoforms activate a pathway that is not shared by p53 and that is required for neuroblastoma cell differentiation in vitro. PMID- 10809759 TI - Classical and nonclassical class I major histocompatibility complex molecules exhibit subtle conformational differences that affect binding to CD8alphaalpha. AB - The cell surface molecules CD4 and CD8 greatly enhance the sensitivity of T-cell antigen recognition, acting as "co-receptors" by binding to the same major histocompatibility complex (MHC) molecules as the T-cell receptor (TCR). Here we use surface plasmon resonance to study the binding of CD8alphaalpha to class I MHC molecules. CD8alphaalpha bound the classical MHC molecules HLA-A*0201, A*1101, -B*3501, and -C*0702 with dissociation constants (K(d)) of 90-220 microm, a range of affinities distinctly lower than that of TCR/peptide-MHC interaction. We suggest such affinities apply to most CD8alphaalpha/classical class I MHC interactions and may be optimal for T-cell recognition. In contrast, CD8alphaalpha bound both HLA-A*6801 and B*4801 with a significantly lower affinity (>/=1 mm), consistent with the finding that interactions with these alleles are unable to mediate cell-cell adhesion. Interestingly, CD8alphaalpha bound normally to the nonclassical MHC molecule HLA-G (K(d) approximately 150 microm), but only weakly to the natural killer cell receptor ligand HLA-E (K(d) >/= 1 mm). Site-directed mutagenesis experiments revealed that variation in CD8alphaalpha binding affinity can be explained by amino acid differences within the alpha3 domain. Taken together with crystallographic studies, these results indicate that subtle conformational changes in the solvent exposed alpha3 domain loop (residues 223-229) can account for the differential ability of both classical and nonclassical class I MHC molecules to bind CD8. PMID- 10809760 TI - Ca2+/calmodulin-dependent kinase II and calcineurin play critical roles in endothelin-1-induced cardiomyocyte hypertrophy. AB - Endothelin-1 (ET-1) induces cardiac hypertrophy. Because Ca(2+) is a major second messenger of ET-1, the role of Ca(2+) in ET-1-induced hypertrophic responses in cultured cardiac myocytes of neonatal rats was examined. ET-1 activated the promoter of the beta-type myosin heavy chain gene (beta-MHC) (-354 to +34 base pairs) by about 4-fold. This activation was inhibited by chelation of Ca(2+) and the blocking of protein kinase C activity. Similarly, the beta-MHC promoter was activated by Ca(2+) ionophores and a protein kinase C activator. beta-MHC promoter activation induced by ET-1 was suppressed by pretreatment with the calmodulin inhibitor, W7, the Ca(2+)/calmodulin-dependent kinase II (CaMKII) inhibitor, KN62, and the calcineurin inhibitor, cyclosporin A. beta-MHC promoter activation by ET-1 was also attenuated by overexpression of dominant-negative mutants of CaMKII and calcineurin. ET-1 increased the activity of CaMKII and calcineurin in cardiac myocytes. Pretreatment with KN62 and cyclosporin A strongly suppressed ET-1-induced increases in [(3)H]phenylalanine uptake and in cell size. These results suggest that Ca(2+) plays a critical role in ET-1 induced cardiomyocyte hypertrophy by activating CaMKII- and calcineurin-dependent pathways. PMID- 10809761 TI - Substitutions of Asn-726 in the active site of yeast DNA topoisomerase I define novel mechanisms of stabilizing the covalent enzyme-DNA intermediate. AB - Eukaryotic DNA topoisomerase I (Top1p) catalyzes changes in DNA topology and is the cellular target of camptothecin. Recent reports of enzyme structure highlight the importance of conserved amino acids N-terminal to the active site tyrosine and the involvement of Asn-726 in mediating Top1p sensitivity to camptothecin. To investigate the contribution of this residue to enzyme catalysis, we evaluated the effect of substituting His, Asp, or Ser for Asn-726 on yeast Top1p. Top1N726S and Top1N726D mutant proteins were resistant to camptothecin, although the Ser mutant was distinguished by a lack of detectable changes in activity. Thus, a basic residue immediately N-terminal to the active site tyrosine is required for camptothecin cytotoxicity. However, replacing Asn-726 with Asp or His interfered with distinct aspects of the catalytic cycle, resulting in cell lethality. In contrast to camptothecin, which inhibits enzyme-catalyzed religation of DNA, the His substituent enhanced the rate of DNA scission, whereas the Asp mutation diminished the enzyme binding of DNA. Yet, these effects on enzyme catalysis were not mutually exclusive as the His mutant was hypersensitive to camptothecin. These results suggest distinct mechanisms of poisoning DNA topoisomerase I may be explored in the development of antitumor agents capable of targeting different aspects of the Top1p catalytic cycle. PMID- 10809762 TI - Hrs-2 regulates receptor-mediated endocytosis via interactions with Eps15. AB - Hrs-2, via interactions with SNAP-25, plays a regulatory role on the exocytic machinery. We now show that Hrs-2 physically interacts with Eps15, a protein required for receptor-mediated endocytosis. The Hrs-2/Eps15 interaction is calcium dependent, inhibited by SNAP-25 and alpha-adaptin, and results in the inhibition of receptor-mediated endocytosis. Immunoelectron microscopy reveals Hrs-2 localization on the limiting membrane of multivesicular bodies, organelles in the endosomal pathway. These data show that Hrs-2 regulates endocytosis, delineate a biochemical pathway (Hrs-2-Eps15-AP2) in which Hrs-2 functions, and suggest that Hrs-2 acts to provide communication between endo- and exocytic processes. PMID- 10809763 TI - Characterization of rapid membrane internalization and recycling. AB - Lipids and other membrane constituents recycle between the plasma membrane and intracellular endocytic compartments. In CHO cells, approximately half of the internalized C(6)-NBD-SM, a fluorescent lipid analogue widely used as a membrane maker, recycles via the endocytic recycling compartment with a t(12) of approximately 12 min (Mayor, S., Presley, J. F., and Maxfield, F. R. (1993) J. Cell Biol. 121, 1257-1269). Surprisingly, the rest returns to the plasma membrane very quickly. A detailed kinetic study presented in this paper indicates that after a brief internalization pulse, 42-62% of the internalized C(6)-NBD-SM returns to the plasma membrane with a t(12) of 1-2 min. Similar results are obtained using HEp2 and nonpolarized Madin-Darby canine kidney cells. Using FM dyes of different hydrophobicity, we show that rapid recycling involves passage through an endocytic organelle that was subsequently identified as the sorting endosome by co-localization with internalized transferrin and low density lipoprotein. These results imply that the membrane internalization rate is much higher than previously estimated, with a t(12) as short as 5-10 min. Rapid internalization and recycling would facilitate processes such as nutrient uptake and cholesterol efflux. PMID- 10809764 TI - Complex interactions between epidermal POU domain and activator protein 1 transcription factors regulate the expression of the profilaggrin gene in normal human epidermal keratinocytes. AB - The human profilaggrin gene is expressed in the granular layer during the late stages of the epidermal differentiation. The proximal promoter region of the gene confers high levels of keratinocyte-specific transcription via interactions with c-Jun/c-Fos heterodimers. Here we provide evidence for another level of complexity in the regulation of the profilaggrin promoter activity. The POU domain proteins Oct1, Skn1a/i, and Oct6, which are abundantly expressed in the epidermal cells, act to both stimulate and repress transcription in a general and a cell type-specific mode. While binding to specific recognition elements within the promoter region, they exert their effects by either stimulating or antagonizing the c-Jun-dependent activity of the promoter. The response of the promoter to forced expression of the POU domain proteins reflects the effect of these transcription factors on the endogenous profilaggrin mRNA synthesis and suggests that the latter requires a fine balance in the amounts and the activities of the individual activator protein 1 and POU domain proteins. PMID- 10809765 TI - Calcium signal transmission between ryanodine receptors and mitochondria. AB - Control of energy metabolism by increases of mitochondrial matrix [Ca(2+)] ([Ca(2+)](m)) may represent a fundamental mechanism to meet the ATP demand imposed by heart contractions, but the machinery underlying propagation of [Ca(2+)] signals from ryanodine receptor Ca(2+) release channels (RyR) to the mitochondria remains elusive. Using permeabilized cardiac (H9c2) cells we investigated the cytosolic [Ca(2+)] ([Ca(2+)](c)) and [Ca(2+)](m) signals elicited by activation of RyR. Caffeine, Ca(2+), and ryanodine evoked [Ca(2+)](c) spikes that often appeared as frequency-modulated [Ca(2+)](c) oscillations in these permeabilized cells. Rapid increases in [Ca(2+)](m) and activation of the Ca(2+)-sensitive mitochondrial dehydrogenases were synchronized to the rising phase of the [Ca(2+)](c) spikes. The RyR-mediated elevations of global [Ca(2+)](c) were in the submicromolar range, but the rate of [Ca(2+)](m) increases was as large as it was in the presence of 30 microm global [Ca(2+)](c). Furthermore, RyR-dependent increases of [Ca(2+)](m) were relatively insensitive to buffering of [Ca(2+)](c) by EGTA. Therefore, RyR-driven rises of [Ca(2+)](m) appear to result from large and rapid increases of perimitochondrial [Ca(2+)]. The falling phase of [Ca(2+)](c) spikes was followed by a rapid decay of [Ca(2+)](m). CGP37157 slowed down relaxation of [Ca(2+)](m) spikes, whereas cyclosporin A had no effect, suggesting that activation of the mitochondrial Ca(2+) exchangers accounts for rapid reversal of the [Ca(2+)](m) response with little contribution from the permeability transition pore. Thus, rapid activation of Ca(2+) uptake sites and Ca(2+) exchangers evoked by RyR-mediated local [Ca(2+)](c) signals allow mitochondria to respond rapidly to single [Ca(2+)](c) spikes in cardiac cells. PMID- 10809766 TI - Colocalization of prostacyclin synthase with prostaglandin H synthase-1 (PGHS-1) but not phorbol ester-induced PGHS-2 in cultured endothelial cells. AB - The subcellular colocalization of prostacyclin synthase (PGIS) with prostaglandin H synthase (PGHS) has not been delineated. To test the hypothesis that its colocalization with PGHS is crucial for prostacyclin synthesis, we determined subcellular locations of PGIS, PGHS-1, and PGHS-2 in bovine aortic endothelial cells by immunofluorescent confocal microscopy. PGIS and PGHS-1 were colocalized to nuclear envelope (NE) and endoplasmic reticulum (ER) in resting and adenovirus infected bovine aortic endothelial cells. PGIS and PGHS-2 were also colocalized to ER in serum-treated or adenovirus-cyclooxygenase-2-infected cells. By contrast, PGIS was not colocalized with PGHS-2 in cells induced with phorbol 12 myristate 13-acetate where PGHS-2 was visualized primarily in vesicle-like structures. The lack of colocalization was accompanied by failed prostacyclin production. Resting ECV304 cells did not produce prostacyclin and had no detectable PGHS-1 and PGIS proteins. Confocal analysis showed abnormal colocalization of PGIS and PGHS-1 to a filamentous structure. Interestingly, the abundant PGIS and PGHS-1 expressed in adenovirus-infected ECV304 cells were colocalized to NE and ER, which synthesized a large quantity of prostacyclin. These findings underscore the importance of colocalization of PGHS and PGIS to ER and NE in prostacyclin synthesis. PMID- 10809767 TI - Structural requirements of heparan sulfate for the binding to the tumor-derived adhesion factor/angiomodulin that induces cord-like structures to ECV-304 human carcinoma cells. AB - Tumor-derived adhesion factor/angiomodulin (AGM) is accumulated in tumor blood vessels and on the endothelial cell surface (Akaogi, K., Okabe, Y., Sato, J., Nagashima, Y., Yasumitsu, H., Sugahara, K., and Miyazaki, K. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 8384-8389). In cell culture, it promotes cell adhesion and morphological changes to form cord-like structures of the human bladder carcinoma cell line ECV-304. The cord formation is prevented by heparin, which inhibits the binding of AGM to ECV-304 cells. This observation suggests that AGM interacts with cell surface heparan sulfate (HS) proteoglycans. In this study, HS glycosaminoglycans and core proteins of integral transmembrane proteoglycans, syndecan-1 and -4, were identified by immunocytochemistry on ECV-304 cells, and the structural requirements for the interaction of HS with AGM were characterized. Inhibition experiments with sulfated polysaccharides and chemically modified heparin derivatives indicated that sulfate groups were essential for both AGM-HS binding and cord-like structure formation and that the rank order of the different sulfate groups in terms of their contribution was N sulfate > 6-O-sulfate > 2-O-sulfate. The minimum size of heparin, a chemical analog of HS, required for the binding to AGM was a dodecasaccharide as determined by competition experiments using size-defined heparin oligosaccharides. Thus, a specific sulfation pattern in the HS of cell surface syndecans of ECV-304 cells is required for AGM binding and the morphological changes. PMID- 10809768 TI - TAJ, a novel member of the tumor necrosis factor receptor family, activates the c Jun N-terminal kinase pathway and mediates caspase-independent cell death. AB - We have isolated a novel member of the TNFR family, designated TAJ, that is highly expressed during embryonic development. TAJ possesses a unique cytoplasmic domain with no sequence homology to the previously characterized members of the TNFR family. TAJ interacts with the TRAF family members and activates the JNK pathway when overexpressed in mammalian cells. Although it lacks a death domain, TAJ is capable of inducing apoptosis by a caspase-independent mechanism. Based on its unique expression profile and signaling properties, TAJ may play an essential role in embryonic development. PMID- 10809769 TI - Gelsolin inhibits apoptosis by blocking mitochondrial membrane potential loss and cytochrome c release. AB - Apoptotic cell death, characterized by chromatin condensation, nuclear fragmentation, cell membrane blebbing, and apoptotic body formation, is also accompanied by typical mitochondrial changes. The latter includes enhanced membrane permeability, fall in mitochondrial membrane potential (Deltapsi(m)) and release of cytochrome c into the cytosol. Gelsolin, an actin regulatory protein, has been shown to inhibit apoptosis, but when cleaved by caspase-3, a fragment that is implicated as an effector of apoptosis is generated. The mechanism by which the full-length form of gelsolin inhibits apoptosis is unclear. Here we show that the overexpression of gelsolin inhibits the loss of Deltapsi(m) and cytochrome c release from mitochondria resulting in the lack of activation of caspase-3, -8, and -9 in Jurkat cells treated with staurosporine, thapsigargin, and protoporphyrin IX. These effects were corroborated in vitro using recombinant gelsolin protein on isolated rat mitochondria stimulated with Ca(2+), atractyloside, or Bax. This protective function of gelsolin, which was not due to simple Ca(2+) sequestration, was inhibited by polyphosphoinositide binding. In addition we confirmed that gelsolin, besides its localization in the cytosol, is also present in the mitochondrial fraction of cells. Gelsolin thus acts on an early step in the apoptotic signaling at the level of mitochondria. PMID- 10809770 TI - Intramolecular interactions between the juxtamembrane domain and phosphatase domains of receptor protein-tyrosine phosphatase RPTPmu. Regulation of catalytic activity. AB - RPTPmu is a receptor-like protein-tyrosine phosphatase (RPTP) whose ectodomain mediates homotypic cell-cell interactions. The intracellular part of RPTPmu contains a relatively long juxtamembrane domain (158 amino acids; aa) and two conserved phosphatase domains (C1 and C2). The membrane-proximal C1 domain is responsible for the catalytic activity of RPTPmu, whereas the membrane-distal C2 domain serves an unknown function. The regulation of RPTP activity remains poorly understood, although dimerization has been proposed as a general mechanism of inactivation. Using the yeast two-hybrid system, we find that the C1 domain binds to an N-terminal noncatalytic region in RPTPmu, termed JM (aa 803-955), consisting of a large part of the juxtamembrane domain (120 aa) and a small part of the C1 domain (33 aa). When co-expressed in COS cells, the JM polypeptide binds to both the C1 and the C2 domain. Strikingly, the isolated JM polypeptide fails to interact with either full-length RPTPmu or with truncated versions of RPTPmu that contain the JM region, consistent with the JM-C1 and JM-C2 interactions being intramolecular rather than intermolecular. Furthermore, we find that large part of the juxtamembrane domain (aa 814-922) is essential for C1 to be catalytically active. Our findings suggest a model in which RPTPmu activity is regulated by the juxtamembrane domain undergoing intramolecular interactions with both the C1 and C2 domain. PMID- 10809771 TI - Normal blood pressure and plasma renin activity in mice lacking the renin-binding protein, a cellular renin inhibitor. AB - In renal extracts, some renin is present as "high molecular weight renin," a heterodimeric complex of renin with the 46-kDa renin-binding protein (RnBP), also known as N-acyl-D-glucosamine 2-epimerase. Because RnBP specifically inhibits renin activity, the protein was proposed to play an important role in the regulation of the renin-angiotensin system (RAS). Using gene targeting, we have generated mice lacking RnBP and tested this hypothesis in vivo. In particular, we analyzed biosynthesis, secretion, and activity of renin and other components of the RAS in mice lacking RnBP. Despite extensive investigations, we were unable to detect any major effects of RnBP deficiency on the plasma and renal RAS or on blood pressure regulation. Contrary to previous hypotheses, we conclude that RnBP does not play a significant role in the regulation of renin activity in plasma or kidney. However, RnBP knockout mice excrete an abnormal pattern of carbohydrates in the urine, indicating a role of the protein in renal carbohydrate metabolism. PMID- 10809772 TI - Distinct Chk2 activation pathways are triggered by genistein and DNA-damaging agents in human melanoma cells. AB - Genistein, a natural isoflavone found in soybeans, exerts a number of biological actions suggesting that it may have a role in cancer prevention. We have previously shown that it potently inhibits OCM-1 melanoma cell proliferation by inducing a G(2) cell cycle arrest. Here we show that genistein exerts this effect by impairing the Cdc25C-dependent Tyr-15 dephosphorylation of Cdk1, as the overexpression of this phosphatase allows the cells to escape G(2) arrest and enter an abnormal chromatin condensation stage. Caffeine totally overrides the genistein-induced G(2) arrest, whereas the block caused by etoposide is not bypassed and that caused by adriamycin is only partially abolished. We also report that genistein activates the checkpoint kinase Chk2 as efficiently as the two genotoxic agents and that caffeine may counteract the activation of Chk2 by genistein but not by etoposide. In contrast, caffeine abolishes the accumulation of p53 caused by all the compounds. Wortmannin does not suppress the Chk2 activation in any situation, suggesting that the ataxia telangiectasia-mutated kinase is not involved in this regulation. Finally, unlike etoposide and adriamycin, genistein induces only a weak response in terms of DNA damage in OCM 1 cells. Taken together, these results suggest that the G(2) checkpoints activated by genistein and the two genotoxic agents involve different pathways. PMID- 10809773 TI - Oxidative stress abolishes leptomycin B-sensitive nuclear export of transcription repressor Bach2 that counteracts activation of Maf recognition element. AB - The mammalian transcription activator Nrf2 plays critical roles in executing oxidative stress response by binding to the regulatory DNA sequence Maf recognition element. Bach2 is an Nrf2-related transcription repressor and a tissue-specific partner of the Maf oncoprotein family. We show here how Bach2 is regulated by an oxidative stress-sensitive conditional nuclear export. In cultured cells, Bach2 was localized in cytoplasm through its C-terminal evolutionarily conserved cytoplasmic localization signal (CLS). The CLS directed leptomycin B-sensitive nuclear export of reporter proteins, suggesting its dependence on the nuclear exporter Crm1/exportin 1. However, the CLS sequence does not bear a resemblance to the leucine-rich class of nuclear export signal, and mutagenesis analysis indicated that a stretch of nonhydrophobic amino acids is essential for its activity. Oxidative stressors aborted the CLS activity and induced nuclear accumulation of Bach2. Whereas oxidative stress is known to activate MARE-dependent transcription, overexpression of Bach2 in cultured cells silenced the inducibility of MARE. The results suggest that Bach2 mediates nucleocytoplasmic communication to couple oxidative stress and transcription repression in mammalian cells. PMID- 10809774 TI - The conformational activation of antithrombin. A 2.85-A structure of a fluorescein derivative reveals an electrostatic link between the hinge and heparin binding regions. AB - Antithrombin is unique among the serpins in that it circulates in a native conformation that is kinetically inactive toward its target proteinase, factor Xa. Activation occurs upon binding of a specific pentasaccharide sequence found in heparin that results in a rearrangement of the reactive center loop removing constraints on the active center P1 residue. We determined the crystal structure of an activated antithrombin variant, N135Q S380C-fluorescein (P14-fluorescein), in order to see how full activation is achieved in the absence of heparin and how the structural effects of the substitution in the hinge region are translated to the heparin binding region. The crystal structure resembles native antithrombin except in the hinge and heparin binding regions. The absence of global conformational change allows for identification of specific interactions, centered on Glu(381) (P13), that are responsible for maintenance of the solution equilibrium between the native and activated forms and establishes the existence of an electrostatic link between the hinge region and the heparin binding region. A revised model for the mechanism of the allosteric activation of antithrombin is proposed. PMID- 10809775 TI - Targeted expression of placental lactogen in the beta cells of transgenic mice results in beta cell proliferation, islet mass augmentation, and hypoglycemia. AB - The factors that regulate pancreatic beta cell proliferation are not well defined. In order to explore the role of murine placental lactogen (PL)-I (mPL-I) in islet mass regulation in vivo, we developed transgenic mice in which mPL-I is targeted to the beta cell using the rat insulin II promoter. Rat insulin II-mPL-I mice displayed both fasting and postprandial hypoglycemia (71 and 105 mg/dl, respectively) as compared with normal mice (92 and 129 mg/dl; p < 0.00005 for both). Plasma insulin concentrations were inappropriately elevated, and insulin content in the pancreas was increased 2-fold. Glucose-stimulated insulin secretion by perifused islets was indistinguishable from controls at 7.5, 15, and 20 mm glucose. Beta cell proliferation rates were twice normal (p = 0. 0005). This hyperplasia, together with a 20% increase in beta cell size, resulted in a 2 fold increase in islet mass (p = 0.0005) and a 1.45-fold increase in islet number (p = 0.0012). In mice, murine PL-I is a potent islet mitogen, is capable of increasing islet mass, and is associated with hypoglycemia over the long term. It can be targeted to the beta cell using standard gene targeting techniques. Potential exists for beta cell engineering using this strategy. PMID- 10809776 TI - Estrogen-modulated estrogen receptor x Pit-1 protein complex formation and prolactin gene activation require novel protein synthesis. AB - Both estrogen receptor (ER) and Pit-1 proteins are essential for the estrogen activated expression of the rat prolactin gene. Our results show that ER.Pit-1 protein complex formation is reduced by estrogen in GH3 and PR1 rat pituitary tumor cells. In the latter, this decrease was blocked by cycloheximide, a protein synthesis inhibitor. On the other hand, the direct addition of estrogen to PR1 cell lysates had no effect on the formation of ER.Pit-1 complexes. Estrogen activated prolactin gene expression was also inhibited by cycloheximide, suggesting that some form of protein synthesis is involved in ER.Pit-1 complex formation and subsequent prolactin gene activation. In support of this notion, we showed that estrogen-induced regulation of ER.Pit-1 complex formation could be transferred from cell lysates prepared from estrogen-treated PR1 cells to control cell lysates. This is not true for GH3 cells; instead, direct administration of estrogen to GH3 cell lysates readily abolished ER.Pit-1 protein complex formation in a dose-dependent manner, and such estrogen-induced regulation was blocked by the antiestrogen ICI 182,780. These findings thus indicate that 1) interaction between ER and Pit-1 proteins is estrogen-regulated in ways specific to different cell types, and 2) auxiliary protein factor synthesis may be involved in this process. PMID- 10809777 TI - Cystine knot mutations affect the folding of the glycoprotein hormone alpha subunit. Differential secretion and assembly of partially folded intermediates. AB - The common glycoprotein hormone alpha-subunit (GPH-alpha) contains five intramolecular disulfide bonds, three of which form a cystine knot motif (10-60, 28-82, and 32-84). By converting each pair of cysteine residues of a given disulfide bond to alanine, we have studied the role of individual disulfide bonds in GPH-alpha folding and have related folding ability to secretion and assembly with the human chorionic gonadotropin beta-subunit (hCG-beta). Mutation of non cystine knot disulfide bond 7-31, bond 59-87, or both (leaving only the cystine knot) resulted in an efficiently secreted folding form that was indistinguishable from wild type. Conversely, the cystine knot mutants were inefficiently secreted (<25%). Furthermore, mutation of the cystine knot disulfide bonds resulted in multiple folding intermediates containing 1, 2, or 4 disulfide bonds. High performance liquid chromatographic separation of intracellular and secreted forms of the folding intermediates demonstrated that the most folded forms were preferentially secreted and combined with hCG-beta. From these studies we conclude that: (i) the cystine knot of GPH-alpha is necessary and sufficient for folding and (ii) there is a direct correlation between the extent of GPH-alpha folding, its ability to be secreted, and its ability to heterodimerize with hCG beta. PMID- 10809778 TI - Frequent occurrence of pre-existing alpha 2-->8-linked disialic and oligosialic acids with chain lengths up to 7 Sia residues in mammalian brain glycoproteins. Prevalence revealed by highly sensitive chemical methods and anti-di-, oligo-, and poly-Sia antibodies specific for defined chain lengths. AB - The pre-existence of alpha2-->8-linked disialic acid (di-Sia) and oligosialic acid (oligo-Sia) structures with up to 7 Sia residues was shown to occur on a large number of brain glycoproteins, including neural cell adhesion molecules (N CAMs), by two highly sensitive chemical methods (Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1998) Anal. Biochem. 261, 191-197; Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1999) Anal. Biochem. 266, 102-109). This unexpected finding was also confirmed using a newly developed antibody prepared using a copolymer of alpha2-->8-linked N-acetylneuraminyl p-vinylbenzylamide and acrylamide as an immunogen and known antibodies whose immunospecificities were determined to be di- and oligo-Sia residues with defined chain lengths. The major significance of the new finding that di- and oligo-Sia chains exist on a large number of brain glycoproteins is 2-fold. First, it reveals a surprising diversity in the number and M(r) of proteins distinct from N-CAM that are covalently modified by these short sialyl glycotopes. Second, it suggests that synthesis of di- and/or oligo-Sia units may be catalyzed by alpha2-->8-sialyltransferase(s) that are distinct from the known polysialyltransferases, STX and PST, which are partially responsible for polysialylation of N-CAM. PMID- 10809779 TI - In vitro studies on tRNA annealing and reverse transcription with mutant HIV-1 RNA templates. AB - The human immunodeficiency virus type 1 (HIV-1) RNA genome encodes a semistable stem-loop structure, the U5-PBS hairpin, which occludes part of the tRNA primer binding site (PBS). In previous studies, we demonstrated that mutations that alter the stability of the U5-PBS hairpin inhibit virus replication. A reverse transcription defect was measured in assays with the virion-extracted RNA-tRNA complexes. We now extend these studies with in vitro synthesized wild-type and mutant RNA templates that were tested in primer annealing and reverse transcription assays. The effect of annealing temperature and the presence of the viral nucleocapsid protein on reverse transcription was analyzed for the templates with a stabilized or destabilized U5-PBS hairpin, and in reactions initiated by tRNA or DNA primers. The results of this in vitro assay are consistent with the in vivo findings, in that both tRNA annealing and initiation of reverse transcription are sensitive to stable template RNA structure. Reverse transcription initiated by a DNA primer is less hindered by secondary structure in the RNA template than tRNA primed reactions. The inhibitory effect of template structure on tRNA-primed reverse transcription is more pronounced in this in vitro assay compared with the in vivo material, indicating that the heat-annealed RNA-tRNA complex differs from the virion-extracted viral RNA-tRNA complex. PMID- 10809780 TI - Elevated cholesterol metabolism and bile acid synthesis in mice lacking membrane tyrosine kinase receptor FGFR4. AB - Heparan sulfate-regulated transmembrane tyrosine kinase receptor FGFR4 is the major FGFR isotype in mature hepatocytes. Fibroblast growth factor has been implicated in the definition of liver from foregut endoderm where FGFR4 is expressed and stimulation of hepatocyte DNA synthesis in vitro. Here we show that livers of mice lacking FGFR4 exhibited normal morphology and regenerated normally in response to partial hepatectomy. However, the FGFR4 (-/-) mice exhibited depleted gallbladders, an elevated bile acid pool and elevated excretion of bile acids. Cholesterol- and bile acid-controlled liver cholesterol 7alpha hydroxylase, the limiting enzyme for bile acid synthesis, was elevated, unresponsive to dietary cholesterol, but repressed normally by dietary cholate. Expression pattern and cholate-dependent, cholesterol-induced hepatomegaly in the FGFR4 (-/-) mice suggested that activation of receptor interacting protein 140, a co-repressor of feed-forward activator liver X receptor alpha, may mediate the negative regulation of cholesterol- and bile acid-controlled liver cholesterol 7alpha-hydroxylase transcription by FGFR4 and cholate. The results demonstrate that transmembrane sensors interface with metabolite-controlled transcription networks and suggest that pericellular matrix-controlled liver FGFR4 in particular may ensure adequate cholesterol for cell structures and signal transduction. PMID- 10809781 TI - Role of Src kinases in the ADAM-mediated release of L1 adhesion molecule from human tumor cells. AB - The ectodomain of certain transmembrane molecules can be released by proteolysis, and the solubilized antigens often exert important biological functions. We demonstrated before that the L1 adhesion molecule is shed from the cell surface. Here we show that L1 release in AR breast carcinoma cells is mediated by a member of the disintegrin metalloproteinase (ADAM) family of proteinases. Up-regulation of L1 shedding by phorbol ester or pervanadate involved distinct mechanisms. Pervanadate induced shedding and rounding-up of cells from the substrate, which was blocked by the Src kinase inhibitor PP2. Tyr phosphorylation of the L1 cytoplasmic tail and the Src kinase Fyn was observed following pervanadate treatment. Up-regulation of L1 release and activation of Fyn occurred also when cells were detached by EDTA suggesting that the regulation of L1 shedding by this pathway was linked to cell morphology and adhesion. The phorbol 12-myristate 13 acetate-induced shedding was inhibited by the protein kinase C inhibitor bisindolylmaleimide I and by PD98059, a specific inhibitor of the mitogen activated protein kinase pathway. Soluble L1 binds to the proteoglycan neurocan and in bound form could support integrin-mediated cell adhesion and migration. We propose that the release of cell-associated adhesion molecules such as L1 may be relevant to promote cell migration. PMID- 10809782 TI - Interaction of two multifunctional proteins. Heterogeneous nuclear ribonucleoprotein K and Y-box-binding protein. AB - The heterogeneous nuclear ribonucleoprotein (hnRNP) K, a component of the hnRNP particles, appears to be involved in several steps of regulation of gene expression. To gain insight into mechanisms of K protein action, we performed two hybrid screens using full-length hnRNP K as a bait. Several novel protein partners were identified, including Y-box-binding protein (YB-1), splicing factors 9G8 and SRp20, DNA-methyltransferase, hnRNP L, and hnRNP U. In vitro binding studies and co-immunoprecipitation from cellular extracts provided evidence for direct interaction between hnRNP K and YB-1. Two distinct domains in YB-1 were responsible for binding to K protein. Each protein was able to transactivate transcription from a polypyrimidine-rich promoter; however, this effect was reduced when K and YB-1 proteins were coexpressed suggesting a functional interaction between these two proteins. PMID- 10809783 TI - Characterization of sPARP-1. An alternative product of PARP-1 gene with poly(ADP ribose) polymerase activity independent of DNA strand breaks. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is an abundant nuclear enzyme that catalyzes the synthesis of poly(ADP-ribose) (pADPr) from its substrate NAD(+) upon binding to DNA strand breaks. Poly(ADP-ribosyl)ation has been implicated in many cellular processes including replication, transcription, and the maintenance of genomic stability. However, studies with mice and cells lacking PARP-1 reveal a critical role for the enzyme in the maintenance of genomic integrity only. Recently, a significant level of poly(ADP-ribose) polymerase activity has been detected in fibroblasts derived from mice lacking PARP-1 following treatment with genotoxic agents (Shieh, W. M., Ame, J-C., Wilson, M. V., Wang, Z-Q., Koh, D. W., Jacobson, M. K., and Jacobson, E. L. (1998) J. Biol. Chem. 273, 30069-30072). We have isolated a cDNA that originates from PARP-1 (-/-) fibroblasts and encodes a polypeptide of 493 amino acid residues bearing poly(ADP-ribose) polymerase activity. This protein, that we named sPARP-1 for short poly(ADP-ribose) polymerase-1, has a calculated mass of 55.3 kDa and is identical in deduced amino acid sequence to the catalytic domain of PARP-1. Radiation hybrid analysis assigned the sPARP-1 gene to the chromosome 1H5-H6 in an immediate proximity to the known location of PARP-1 gene, indicating that sPARP-1 and PARP-1 are most probably products of the same gene. Active sPARP-1 is present in both PARP-1 (+/+) and PARP-1 (-/-) cells as demonstrated by activity-Western blotting and immunostaining using a specific antibody developed against sPARP-1. Like PARP-1, sPARP-1 is localized in the cell nucleus, uses NAD(+) as a substrate and is inhibited by nicotinamide analogues. sPARP-1 produces pADPr of similar length and structure to that of PARP-1. However, contrary to PARP-1, sPARP-1 does not require DNA strand breaks for its activation, although it is stimulated following genotoxic treatments. PMID- 10809784 TI - 120- and 160-kDa receptors for endogenous mitogenic peptide, phytosulfokine alpha, in rice plasma membranes. AB - Plant cells in culture secrete a sulfated peptide named phytosulfokine-alpha (PSK alpha), and this peptide induces the cell division and/or cell differentiation by means of specific high and low affinity receptors. Putative receptor proteins for this autocrine type growth factor were identified by photoaffinity labeling of plasma membrane fractions derived from rice suspension cells. Incubation of membranes with a photoactivable (125)I-labeled PSK-alpha analog, [N(epsilon)-(4 azidosalicyl)Lys(5)]PSK-alpha (AS-PSK-alpha), followed by UV irradiation resulted in specific labeling of 120- and 160-kDa bands in SDS-polyacrylamide gel electrophoresis. The labeling of both bands was completely inhibited by unlabeled PSK-alpha and partially decreased by PSK-alpha analogs possessing moderate binding activities. In contrast, PSK-alpha analogs that have no biological activity showed no competition for (125)I-AS-PSK-alpha binding, confirming the specificity of binding proteins. Analysis of the affinity of (125)I incorporation into the protein by ligand saturation experiments gave apparent K(d) values of 5.0 nm for the 120-kDa band and 5.4 nm for the 160-kDa band, suggesting that both proteins correspond to the high affinity binding site. Treatment of (125)I-AS-PSK alpha cross-linked proteins with peptide N-glycosidase F demonstrated that both proteins contained approximately 10 kDa of N-linked oligosaccharides. Specific cross-linking of (125)I-AS-PSK-alpha was also observed by using plasma membranes derived from carrot and tobacco cells, indicating the widespread occurrence of the binding proteins. Together, these data suggest that the 120- and 160-kDa proteins are PSK-alpha receptors that mediate the biological activities of PSK alpha. PMID- 10809785 TI - ATP modulates subunit-subunit interactions in an ATP-binding cassette transporter (MalFGK2) determined by site-directed chemical cross-linking. AB - The binding protein-dependent maltose transport system of enterobacteria (MalFGK(2)), a member of the ATP-binding cassette (ABC) transporter superfamily, is composed of two integral membrane proteins, MalF and MalG, and of two copies of an ATPase subunit, MalK, which hydrolyze ATP, thus energizing the translocation process. In addition, an extracellular (periplasmic) substrate binding protein (MalE) is required for activity. Ligand translocation and ATP hydrolysis are dependent on a signaling mechanism originating from the binding protein and traveling through MalF/MalG. Thus, subunit-subunit interactions in the complex are crucial to the transport process but the chemical nature of residues involved is poorly understood. We have investigated the proximity of residues in a conserved sequence ("EAA" loop) of MalF and MalG to residues in a helical segment of the MalK subunits by means of site-directed chemical cross linking. To this end, single cysteine residues were introduced into each subunit at several positions and the respective malF and malG alleles were individually co-expressed with each of the malK alleles. Membrane vesicles were prepared from those double mutants that contained a functional transporter in vivo and treated with Cu(1,10-phenanthroline)(2)SO(4) or bifunctional cross-linkers. The results suggest that residues Ala-85, Lys-106, Val-114, and Val-117 in the helical segment of MalK, to different extents, participate in constitution of asymmetric interaction sites with the EAA loops of MalF and MalG. Furthermore, both MalK monomers in the complex are in close contact to each other through Ala-85 and Lys 106. These interactions are strongly modulated by MgATP, indicating a structural rearrangement of the subunits during the transport cycle. These data are discussed with respect to current transport models. PMID- 10809786 TI - The Yap1p-dependent induction of glutathione synthesis in heat shock response of Saccharomyces cerevisiae. AB - Glutathione is synthesized in two sequential reactions catalyzed by gamma glutamylcysteine synthetase (GSH1 gene product) and glutathione synthetase (GSH2 gene product). The expression of GSH1 in Saccharomyces cerevisiae has been known to be up-regulated by Yap1p, a critical transcription factor for the oxidative stress response in yeast. The present study demonstrates that GSH2 expression is also regulated by Yap1p under oxidative stress-induced conditions. In addition to oxidative stress, expression of GSH1 and GSH2 was induced by heat shock stress in a Yap1p-dependent manner with subsequent increases in intracellular glutathione content. Oxygen respiration rate increased when cells were exposed to higher temperatures, and as a result, intracellular oxidation levels were increased. The heat shock-induced expression of GSH1 and GSH2 did not occur under anaerobic conditions. Furthermore, even under aerobic conditions, the heat shock response of these genes was not observed when cells were pretreated with KCN to block oxygen respiration. We speculate that heat shock stress enhances oxygen respiration, which in turn results in an increase in the generation of reactive oxygen species in mitochondria. This signal may be mediated by Yap1p, resulting in the elevation of intracellular glutathione levels. PMID- 10809787 TI - High affinity binding of beta 2-glycoprotein I to human endothelial cells is mediated by annexin II. AB - Beta(2)-glycoprotein I (beta(2)GPI) is an abundant plasma phospholipid-binding protein and an autoantigen in the antiphospholipid antibody syndrome. Binding of beta(2)GPI to endothelial cells targets them for activation by anti-beta(2)GPI antibodies, which circulate and are associated with thrombosis in patients with the antiphospholipid antibody syndrome. However, the binding of beta(2)GPI to endothelial cells has not been characterized and is assumed to result from association of beta(2)GPI with membrane phospholipid. Here, we characterize the binding of beta(2)GPI to endothelial cells and identify the beta(2)GPI binding site. (125)I-beta(2)GPI bound with high affinity (K(d) approximately 18 nm) to human umbilical vein endothelial cells (HUVECs). Using affinity purification, we isolated beta(2)GPI-binding proteins of approximately 78 and approximately 36 kDa from HUVECs and EAHY.926 cells. Amino acid sequences of tryptic peptides from each of these were identical to sequences within annexin II. A role for annexin II in binding of beta(2)GPI to cells was confirmed by the observations that annexin II-transfected HEK 293 cells bound approximately 10-fold more (125)I beta(2)GPI than control cells and that anti-annexin II antibodies inhibited the binding of (125)I-beta(2)GPI to HUVECs by approximately 90%. Finally, surface plasmon resonance studies revealed high affinity binding between annexin II and beta(2)GPI. These results demonstrate that annexin II mediates the binding of beta(2)GPI to endothelial cells. PMID- 10809788 TI - Somatostatin interferes with thyrotropin-induced G1-S transition mediated by cAMP dependent protein kinase and phosphatidylinositol 3-kinase. Involvement of RhoA and cyclin E x cyclin-dependent kinase 2 complexes. AB - cAMP-mediated cell proliferation is a complex process that involves multiple pathways. Using a cAMP-dependent cell system, FRTL-5 thyroid cells, we have previously demonstrated the existence of a precise autocrine loop in the control of cell proliferation that involves the positive effector thyrotropin (TSH) and the general inhibitor somatostatin. In search of the regulatory mechanisms responsible for the TSH and somatostatin control of cell proliferation, we analyzed the cell cycle regulatory proteins and the cellular pathways involved in the action of both signals. The results show that specific inhibition of cAMP dependent protein kinase (PKA) and phosphatidylinositol (PI) 3-kinase blocks independently TSH-induced FRTL-5 cell proliferation and that somatostatin interferes with both signals. Each pathway activates different proteins required for G(1)/S progression. Thus, PKA is responsible for the TSH-induction of 3 hydroxy-3-methylglutaryl-CoA reductase mRNA levels, RhoA activation, and down regulation of p27(kip1). These correlated events are necessary for FRTL-5 cell proliferation after TSH stimulation. Moreover, TSH through PKA pathway increases cyclin-dependent kinase 2 levels, whereas PI 3-kinase signaling increases cyclin E levels. Together, both pathways finally converge, increasing the formation and activation of cyclin E x cyclin-dependent kinase 2 complexes and the phosphorylation of the retinoblastoma protein, two important steps in the transition from G(1) to S phase in growth-stimulated cells. Somatostatin exerts its antiproliferative effect inhibiting more upstream the TSH stimulation of PKA and PI 3-kinase, interfering with the TSH-mediated increases of intracellular cAMP levels by inactivation of adenylyl cyclase activity. Together, these results suggest the existence of a PKA-dependent pathway and a new PKA-independent PI 3 kinase pathway in the TSH/cAMP-mediated proliferation of FRTL-5 thyroid cells. PMID- 10809789 TI - ACE inhibitors and AT(1) receptor antagonists-beyond the haemodynamic effect. PMID- 10809790 TI - Use of corticosteroids in nephrology - risk and prevention of osteoporosis induction. PMID- 10809791 TI - Great strides in the understanding of renal magnesium and calcium reabsorption. PMID- 10809792 TI - Clostridium difficile in patients with renal failure - management of an outbreak using biotherapy. PMID- 10809793 TI - Blood monocyte phenotypes and soluble endotoxin receptor CD14 in systemic inflammatory diseases and patients with chronic renal failure. PMID- 10809794 TI - Water quality and complications of haemodialysis. PMID- 10809795 TI - Calcium channel blockers - the effect on renal changes in clinical and experimental diabetes: an overview. PMID- 10809796 TI - G(-699)/C polymorphism in the bradykinin-1 receptor gene in patients with renal failure. AB - BACKGROUND: Bradykinin is thought to have protective effects on the progression of renal failure. Of particular interest, it has been reported that one polymorphism in the promoter region of the human kinin B1-receptor gene which is associated with higher activity, is less frequently found in patients with end stage renal failure. The present study was performed to independently confirm these results. DESIGN: Cross-sectional study on 376 healthy controls, 262 non diabetic dialysis patients and 175 patients with type 1 diabetes >/=10 years and microalbuminuria (of whom 21 were dialysis-dependent) and 334 patients with type 2 diabetes >/=10 years and nephropathy (of whom 61 were dialysis-dependent). METHODS: Genotyping was performed by polymerase chain reaction, followed by restriction enzyme analysis. RESULTS: All groups were in Hardy Weinberg equilibrium. The study showed no significant difference in the frequency of the C allele between controls (0.093) and non-diabetic dialysis patients (0.095). No significant difference in C-allele frequency was observed between controls and patients with type 1 diabetes and microalbuminuria (0.092) or patients with type 2 diabetes and nephropathy (0.099). CONCLUSION: In large cohorts of patients with non-diabetic end-stage renal disease and diabetic renal disease with and without end-stage renal failure, no change in the frequency of the C-699 allele of the B 1-receptor gene was found. PMID- 10809797 TI - Glial cell line-derived neurotrophic factor (GDNF) is expressed in the human kidney and is a growth factor for human mesangial cells. AB - BACKGROUND: Glial cell line-derived neurotrophic factor (GDNF), a recently cloned member of the transforming growth factor-beta (TGF-beta) superfamily, is a potent neurotrophic factor in vitro and in vivo. GDNF is essential for nephrogenesis and the highest expression of GDNF is found in the developing kidney. Increased plasma GDNF levels have recently been documented in patients with chronic renal failure; the source and role of this increase, however, remain unclear. No data are available about the expression of GDNF in human adult kidney or human adult mesangial cell (HMC) cultures. We hypothesized that GDNF, similar to other members of the TGF-beta superfamily, might play a role as a growth factor in the pathogenesis of glomerulosclerosis. METHODS: To address this hypothesis, we first investigated (by RT-PCR) the expression of GDNF mRNA and the mRNAs of the GDNF receptors Ret and GFRalpha-1 in (i) adult human renal cortex and medulla and (ii) in HMC in culture. The results were compared to the expression of these molecules in different developmental stages of the rat kidney. We found that both GDNF and its receptors were expressed in human adult kidney and HMC. Since this finding implicates a role for GDNF beyond nephrogenesis, i.e. in renal physiology/pathophysiology, we investigated the effect of GDNF on HMC growth, i.e. (i) cellular protein synthesis as an index of hypertrophy ([(3)H]methionine incorporation), (ii) DNA synthesis ([(3)H]thymidine incorporation) and cell proliferation (cell numbers) as indices of hyperplasia, and (iii) extracellular matrix synthesis, i.e. collagenous and non-collagenous extracellular proteins ([(3)H]proline incorporation into the collagenase-sensitive and -insensitive fraction). HMC cultures were used as a surrogate model for the development of glomerulosclerosis. RESULTS: GDNF induced a biphasic growth stimulatory effect in HMC with stimulation at the lowest concentration used (2 ng/ml) but had no effect at higher concentrations (20 and 50 ng/ml). In contrast, cellular protein synthesis and extracellular matrix synthesis were significantly and dose dependently increased by GDNF. CONCLUSIONS: These results suggest that GDNF, similar to other members of the TGF-beta superfamily, might play a role as a growth factor for mesangial cells and might thus be a player in the pathogenesis of glomerulosclerosis. PMID- 10809798 TI - TGF-beta(1) down-regulates inflammatory cytokine-induced VCAM-1 expression in cultured human glomerular endothelial cells. AB - BACKGROUND: Endothelial cells are active participants in the processes controlling coagulation, inflammation and the immune response. Variations are recognized between endothelia isolated from different vascular beds as well as from different species. Though transforming growth factor-beta(1) (TGF-beta(1)) has been known to have an anti-inflammatory action, little is known about its effect on expression of cellular adhesion molecules during the inflammatory process in human glomerular endothelial cells. The aim of this study was to determine the effect of TGF-beta(1) on the inflammatory cytokine-induced expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured human glomerular endothelial cells. METHODS: The culture of human glomerular endothelial cells was established using the normal portion of nephrectomized renal tissues and identified by factor VIII staining and cellular uptake of fluorescent-labelled acetylated low-density lipoprotein (LDL). The endothelial cells were stimulated by interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha) and interferon-gamma (IFN-gamma) with or without TGF-beta(1). Cellular expression of VCAM-1 was measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry, and VCAM-1 mRNA was measured by Northern blot analysis. RESULTS: TGF-beta(1) (1, 10 and 25 ng/ml) blunted IL-1beta- (5 ng/ml) induced VCAM-1 expression significantly (OD=1.08+/-0.14, 1. 10+/-1.16 and 1.05+/ 0.14 vs IL-1beta=1.97+/-0.29, n=6, P<0.05) in ELISA. The addition of TGF-beta(1) (1, 10 and 25 ng/ml) also suppressed TNF-alpha- (10 ng/ml) induced VCAM-1 expression (OD=1. 14+/-0.15, 1.17+/-0.17 and 1.18+/-0.16 vs TNF-alpha=1.96+/ 0.26, n=6, P<0.05). The same results were obtained by flow cytometry. TGF-beta(1) (10 ng/ml) inhibited both IL-1beta- (5 ng/ml) and TNF-alpha-(10 ng/ml) induced expression of VCAM-1 (MFI: IL-1beta=90. 8+/- 17.6, IL-1beta+TGF-beta(1)=37.8+/ 14.9, TNF-alpha=113.6+/- 12.4, TNF-alpha+TGF-beta(1)=64.3+/-13.8, mean+/-SD, n=3, P<0.05). By Northern blot analysis, TGF-beta(1) (10 ng/ml) significantly suppressed the stimulatory effect of IL-1beta and TNF-alpha. CONCLUSIONS: These results show that TGF-beta(1) down-regulates the inflammatory cytokine-induced expression of VCAM-1 in human glomerular endothelial cells, which could be a novel mechanism for the anti-inflammatory action of TGF-beta(1) during the inflammatory processes in human glomerular diseases. PMID- 10809799 TI - Hypomagnesaemia-hypercalciuria-nephrocalcinosis: a report of nine cases and a review. AB - BACKGROUND: The cardinal characteristics of primary hypomagnesaemia hypercalciuria-nephrocalcinosis include renal magnesium wasting, marked hypercalciuria, renal stones, nephrocalcinosis, a tendency towards chronic renal insufficiency and sometimes even ocular abnormalities or hearing impairment. METHODS: As very few patients with this syndrome have been described, we provide information on nine patients on follow-up at our institutions and review the 42 cases reported in the literature (33 females and 18 males). RESULTS: Urinary tract infections, polyuria-polydipsia, renal stones and tetanic convulsions were the main clinical findings at diagnosis. The clinical course was highly variable; renal failure was often reported. The concomitant occurrence of ocular involvement or hearing impairment was reported in a large subset of patients. Parental consanguinity was noted in some families. CONCLUSIONS: The results indicate an autosomal recessive inheritance. The diagnosis of primary hypomagnesaemia-hypercalciuria-nephrocalcinosis deserves consideration in any patient with nephrocalcinosis and hypercalciuria. PMID- 10809800 TI - Wegener's granulomatosis: clinical course in 108 patients with renal involvement. AB - BACKGROUND: The aim of this study was to evaluate the clinical course of patients with Wegener's granulomatosis and renal involvement, with special reference to relapse rate, renal and patient survival and morbidity from serious infections. METHODS: A retrospective analysis was carried out of 108 patients presenting with Wegener's granulomatosis and active renal disease in eight hospitals in Norway between 1988 and 1998. Multivariate analysis was used to investigate whether selected variables predicted relapse, renal and patient survival and serious infections. RESULTS: Median follow-up was 41.5 months. Twenty-two patients (20.4%) were admitted with a need for dialysis. Complete remission was obtained in 81.5% after a median of 4 months, and 54.7% relapsed after a median of 22. 5 months. Two- and five-year renal survival was 86 and 75%, respectively, and 22.8% developed end-stage renal disease (ESRD). Two- and five-year patient survival was 88 and 74%, respectively, and the cumulative mortality was 3.8 times higher than expected. The relative risk of relapse increased with the use of intravenous pulse cyclophosphamide compared with daily oral cyclophosphamide. Initial renal function predicted renal survival, and low serum albumin and high age at treatment start increased the mortality risk. Thirty one per cent of the patients were hospitalized for serious infections during follow-up. Old age increased the risk of having an infection. CONCLUSIONS: The current treatment of Wegener's granulomatosis does not prevent relapse, development of ESRD and serious treatment-induced infections in a considerable fraction of the patients. Alternative strategies for the management of this disease will be an important objective for further studies. PMID- 10809801 TI - Synthesis of immunoglobulins against Haemophilus parainfluenzae by tonsillar lymphocytes from patients with IgA nephropathy. AB - BACKGROUND: We previously demonstrated glomerular deposition of Haemophilus parainfluenzae (HP) antigens and the presence of IgA antibody against HP antigens in patients with IgA nephropathy (IgAN). In this report we examine the synthesis of immunoglobulins against HP antigens in tonsillar lymphocytes from patients with IgAN. METHODS: We used tonsillar lymphocytes isolated from the palatine tonsils of 15 patients with IgAN and 16 patients with chronic tonsillitis but without renal disease. We examined lymphocyte proliferation and production of IgA, IgG, and IgM antibodies against HP antigens by measuring thymidine uptake and concentrations of these antibodies in culture supernatants after lymphocyte incubation with HP antigens by ELISA. RESULTS: Lymphocytes from patients with IgAN showed a significantly higher stimulation index (SI) on exposure to HP antigens (thymidine incorporation in tonsillar lymphocytes exposed to HP (c.p.m.)/ thymidine incorporation in unstimulated tonsillar lymphocytes (c.p.m.)) than did controls (P=0. 0015). Lymphocytes from patients with IgAN also showed a significantly higher IgA SI (concentrations of IgA against HP antigens in supernatants from HP-stimulated lymphocytes/IgA against HP antigens in supernatants from unstimulated tonsillar lymphocytes) than did controls (P=0.0004). We found positive correlations between concentrations of IgA and IgG antibodies, between IgA and IgM antibodies, and between IgG and IgM antibodies against HP antigens after HP stimulation. CONCLUSIONS: Our results suggest that HP antigens stimulate tonsillar T and B lymphocytes in patients with IgAN and that these patients have polyclonal activation of lymphocytes against HP antigens, with isotype switching of antibody production from IgM to IgA. PMID- 10809802 TI - Plasminogen activator inhibitor-1 and apolipoprotein E gene polymorphisms and diabetic angiopathy. AB - BACKGROUND: A point mutation in the plasminogen activator inhibitor-1 (PAI-1) gene and a three-allelic variation in the apolipoprotein-E (ApoE) gene have been suggested as risk factors for the development of diabetic micro- and macrovascular complications. METHODS: We studied 198 type 1 diabetic patients with diabetic nephropathy [121 men, age (mean+/-SD) 41+/-10 years, diabetes duration 28+/-8 years] and 192 patients with persistent normoalbuminuria (118 men, age 43+/-10 years, diabetes duration 27+/-9 years). RESULTS: Male patients with nephropathy had elevated plasma PAI-1 levels [geometric mean (95% CI)], 70 (62-79) ng/ml, compared with normoalbuminuric men, 43 (38-47) ng/ml, P<0.001. Even though nephropathic patients with the 4G4G genotype tended to have higher plasma PAI-1 levels, P=0.06, no difference in allele frequency (4G/5G) was seen between patients with and without nephropathy: 0.538/0.462 vs 0.539/0.461, respectively. Nor did ApoE allele frequencies (epsilon2/epsilon3/epsilon4) differ between nephropathic and normoalbuminuric patients: 0.099/0.749/0. 152 vs 0.081/0.745/0.174, respectively. Genotype distributions were also similar, n.s. Coronary heart disease was more prevalent (36%) among nephropathic patients carrying the atherogenic epsilon4-allele compared with 12% in patients with the epsilon3,epsilon3 genotype, P<0.001. No associations between diabetic retinopathy and PAI-1 or ApoE polymorphisms were observed, n.s. CONCLUSIONS: The ApoE polymorphism may accelerate the development of coronary heart disease often seen in Caucasian patients with type 1 diabetes and diabetic nephropathy, a condition characterized by elevated plasma PAI-1 in men. Neither the PAI-1 nor the ApoE gene polymorphism contributes to the genetic susceptibility to diabetic nephropathy or retinopathy. PMID- 10809803 TI - Relationship of renal dysfunction to proximal arterial disease severity in atherosclerotic renovascular disease. AB - BACKGROUND: Renal impairment is common in patients with atherosclerotic renovascular disease (ARVD), but its pathogenesis is uncertain. This study investigated whether any relationship existed between renal function and the severity of proximal renal arterial lesions in patients with ARVD. METHODS: A cohort of 71 patients had creatinine clearance measured at the time of digital subtraction angiography; eight patients were diabetics and were excluded from further analysis. The severity of proximal renovascular lesions was estimated by standard methodology, and patients were sub-grouped according to residual patency of the proximal renal arteries (e.g. normal=2.0; unilateral occlusion )RAO(=1.0). Renal bipolar lengths at ultrasound were also assessed. RESULTS: Sixty-three non diabetic patients (mean+/-SD age 67.7+/-5.8 years; 34 males) were suitable for study. No differences in renal function (mean+/-SD creatinine clearance (ml/min)) were seen between patients with unilateral (32. 1+/-18.9, n=36) or bilateral (31.7+/-20.9, n=27) disease, or between sub-groups with RAS <60% (28.3+/-13.9, n=15), unilateral RAS >60% (38.9+/-24.6, n=12), bilateral RAS >60% (36.3+/-20.4, n=6) or unilateral RAO (30.3+/-17.7, n=28), and mean average renal size similarly did not differ between the sub-groups. No correlation existed between residual patency and creatinine clearance (r=0.015); mean+/-SD renal function was almost identical in the four patency sub-groups, and average renal size mirrored this pattern. Mean 24-h urinary protein excretion was similar for the four groups, but patients with minimal ARVD had significantly less comorbid vascular disease. CONCLUSIONS: These findings suggest that the severity of proximal renal artery lesions is often unrelated to the severity of renal dysfunction in patients with ARVD. Associated renal parenchymal damage is the more probable arbiter of renal dysfunction, and this should be considered when revascularization procedures are contemplated. PMID- 10809804 TI - Renal and systemic effects of continued treatment with renin inhibitor remikiren in hypertensive patients with normal and impaired renal function. AB - BACKGROUND: Remikiren is an orally active renin inhibitor with established antihypertensive efficacy. As a single dose it induces renal vasodilatation, suggesting specific renal actions. Data on the renal effects of continued treatment by renin inhibition are not available, either in subjects with normal, or in subjects with impaired renal function. METHODS: The effect of 8 days of treatment with remikiren 600 mg o.i.d. on blood pressure, renal haemodynamics, and proteinuria was studied in 14 hypertensive patients with normal or impaired renal function.The study was conducted on an ambulatory in-hospital basis and was designed in a single-blind, longitudinal order. RESULTS: Remikiren induced a significant peak fall in mean arterial pressure of 11.2+/-0.8%, with corresponding trough values of -6+/-0.8%. This fall was somewhat more pronounced in the patients with renal function impairment (-13.3 vs -9.6%; P<0.01). Glomerular filtration rate remained stable, whereas effective renal plasma flow increased from 301+/-35 to 330+/-36 ml/min/1.73 m(2) (P<0.05). Filtration fraction and renal vascular resistance fell by 10+/-2% and 15+/-2% respectively (both P<0.01). Remikiren induced a cumulated sodium loss of -82+/-22 mmol and a positive potassium balance of 49+/-9 mmol (both P<0.01). During remikiren, proteinuria fell by 27% (range -18 to -38%; P<0.01) in the patients with overt proteinuria at onset (n=6). In the remainder of the patients albuminuria fell by 20% (range -1 to -61%, P<0.05). No side-effects were observed. CONCLUSIONS: Continued treatment with remikiren induced a sustained fall in blood pressure, renal vasodilatation, negative sodium balance, and a reduction in glomerular protein leakage. These data are consistent with a renoprotective potential of renin inhibition. PMID- 10809806 TI - Predictors of hospitalization and death among pre-dialysis patients: a retrospective cohort study. AB - BACKGROUND: Although there is abundant research describing predictors of patient morbidity and mortality among dialysis patients, predictors of adverse clinical outcomes among pre-dialysis patients are less well defined. The purpose of this study was to identify baseline predictors of first non-elective hospitalization among a retrospective cohort of 362 pre-dialysis patients. METHODS: Univariate and multivariate Cox proportional hazard models were used to identify predictors of hospitalization prior to dialysis initiation, adjusted for baseline creatinine level. Dialysis initiation, loss to follow-up, and study conclusion were censored events. Secondary outcomes included cause-specific hospitalization and death. RESULTS: Univariate analysis indicated that advanced age (RR 1.026, CI 1.016 1.037), number of prescribed anti-hypertensive medications (RR 1.149, CI 1.019 1.296), history of myocardial infarction (RR 1.979, CI 1.339-2.926), congestive heart failure (RR 2.299, CI 1.616-3.270), angina (RR 2.289, CI 1.695-3.091), peripheral vascular disease (RR 1.841, CI 1.282-2.644), renal failure secondary to nephrosclerosis (RR 1.413, CI 1.033-1.933) or renal artery stenosis (RR 1.587, CI 1.036-2.430), lower baseline haemoglobin level (RR 0.986, CI 0.979-0.992), and baseline creatinine greater than 300 micromol/l (RR 1.636, CI 1.233-2.171) were predictors of hospitalization. Gender, diabetes, diastolic blood pressure, mean arterial pressure, history of stroke, and hypoalbuminaemia did not predict outcome. Multivariate analysis, adjusted for baseline creatinine level, selected advanced age (RR 1. 017, CI 1.006-1.027), angina (RR 1.893, CI 1.371-2.613), peripheral vascular disease (RR 1.545, CI 1.054-2.266), and haemoglobin level (RR 0.987, CI 0.944-0.979) as independent predictors of hospitalization. CONCLUSION: Advanced age, co-morbid cardiovascular illness and anaemia are independent predictors of non-elective hospitalization prior to dialysis initiation. Further study is needed to determine the extent to which aggressive pre-dialysis management of anaemia and cardiovascular disease can improve patient outcomes. PMID- 10809805 TI - Reduced secretion of proinflammatory cytokines of monosodium urate crystal stimulated monocytes in chronic renal failure: an explanation for infrequent gout episodes in chronic renal failure patients? AB - BACKGROUND: In gouty arthritis, monosodium urate (MSU) crystals interact with monocytes and neutrophils to produce inflammatory reactions associated with acute synovitis. In patients with end-stage renal disease (ESRD), gouty arthritis is a rare condition despite often severe hyperuricaemia. We wondered whether differences in the secretion of proinflammatory cytokines by MSU crystal stimulated monocytes might be one explanation for the low incidence of gouty arthritis in patients with ESRD compared with healthy controls. METHODS: Thirteen patients with ESRD on intermittent haemodialysis treatment, six patients with chronic renal failure not yet on dialysis, and 15 age- and sex-matched healthy controls were examined. Monocytes, purified from peripheral blood mononuclear cells (PBMC) by immunomagnetic bead separation, were incubated for 18 h in the presence of MSU crystals, Escherichia coli lipopolysaccharide (LPS) or medium alone. The supernatants were studied for the presence of interleukin (IL)-1beta, IL-6 and tumour necrosis factor-alpha (TNF-alpha) using cytokine-specific enzyme linked immunosorbent assays. RESULTS: Monocytes from patients with ESRD produced significantly lower amounts of IL-1beta, IL-6 and TNF-alpha after stimulation with MSU crystals or LPS than did monocytes from healthy subjects. Cytokine production was not significantly different between ESRD patients on haemodialysis and chronic renal failure patients not yet on dialysis. Artificial MSU crystals were stronger stimuli than tophus-derived 'natural' MSU crystals. CONCLUSION: We demonstrate that monocyte-associated immunosuppression in ESRD leads to reduced secretion of proinflammatory cytokines in response to stimuli such as MSU crystals. This may be one of the factors preventing many ESRD patients from the manifestation of acute gout despite often severe hyperuricaemia. PMID- 10809807 TI - A new assay method that detects only intact osteocalcin. Two-step non-invasive diagnosis to predict adynamic bone disease in haemodialysed patients. AB - BACKGROUND: We studied the usefulness of a new assay method that detects only the intact human osteocalcin molecule in haemodialysed patients. METHODS: Iliac bone biopsy specimens obtained from 62 haemodialysed patients were analysed. RESULTS: Bone formation rates (BFR/BS) correlated positively with serum intact osteocalcin concentrations (n=62), osteocalcin concentrations assayed by a conventional method (n=31), parathyroid hormone (PTH) concentrations (n=62), and total alkaline phosphatase concentrations (r=0.602, 0. 588, 0.650, and 0.401 respectively). Based on ROC curve and Youden index analysis, the optimal cut-off value to distinguish adynamic bone disease from a mild lesion was 195 pg/ml of serum PTH concentration (Youden index=0.233) or 30 ng/ml of serum intact osteocalcin concentration (Youden index=0.628). The optimal cut-off value to distinguish between hyperparathyroid bone and a mild lesion was 455 pg/ml of serum PTH level (Youden index=0.63) or 50 ng=ml of serum intact osteocalcin concentration (Youden index=0.634). Since both ROC curve and Youden index suggested that the serum PTH concentration was not a good marker to distinguish adynamic bone from a mild lesion or hyperparathyroid bone, we designed a two-step procedure. The first step was the diagnosis of adynamic bone (cut-off: 65 pg/ml) or hyperparathyroid bone (cut-off: 455 pg/ml) according to serum PTH concentration. In a second step, we assessed serum intact osteocalcin concentration in patients with serum PTH concentrations between 65 and 455 pg/ml. The cut-off values for adynamic and hyperparathyroid bone in this diagnostic approach were 30 and 70 ng/ml respectively. As a result, 49 out of 62 patients were diagnosed properly. The Youden index of this two-step diagnosis was 0.527 and 0.661 for adynamic bone and hyperparathyroid bone respectively. Sensitivity markedly improved to 94.4% and 96.2% for adynamic bone and hyperparathyroid bone respectively, without sacrificing specificity (84.0 and 88.8% respectively). CONCLUSION: Measurement of serum intact osteocalcin concentration is useful for the diagnosis of adynamic bone in haemodialysed patients. A two-step procedure involving also simultaneous measurement of serum PTH concentration further improved the sensitivity of each individual marker while maintaining specificity. PMID- 10809808 TI - Low free thyroxine concentrations and deficient nocturnal surge of thyroid stimulating hormone in haemodialysed patients compared with undialysed patients. AB - BACKGROUND: There is little information on the differences in pituitary-thyroid function between undialysed and haemodialysed patients. METHODS: Serum concentrations of free thyroxine (T(4)) and free triiodothyronine (T(3)), measured by enhanced chemiluminescence immunoassay, and thyroid-stimulating hormone (TSH) were compared in undialysed (n=22) and haemodialysed patients (n=85). The response of the serum TSH concentration to exogenously administered thyrotropin-releasing hormone (TRH) and circadian variation in serum TSH were also studied in the two groups. RESULTS: Serum free T(4) concentration was significantly lower in haemodialysed than in undialysed patients (1.02+/-0.02 vs 1.33+/-0.06 ng/dl, P<0.0001). Serum concentrations of free T(3) and TSH were essentially the same for the two groups. The response of serum TSH concentration to TRH was basically the same. Serum TSH concentration in undialysed patients during the night and in the morning were 142.4+/-15.4% and 121.7+/-4.1% of that during the day, the differences being significantly different. A nocturnal surge of TSH was not observed in haemodialysed patients. CONCLUSIONS: Low serum free T(4) concentration and a deficient nocturnal surge of TSH were found in haemodialysed patients compared with undialysed patients. The deficient nocturnal surge of TSH may contribute to the lower serum free T(4) concentration in haemodialysed patients. PMID- 10809809 TI - Variability of relative blood volume during haemodialysis. AB - BACKGROUND: A decrease in blood volume is thought to play a role in dialysis related hypotension. Changes in relative blood volume (RBV) can be assessed by means of continuous haematocrit measurement. We studied the variability of RBV changes, and the relation between RBV and ultrafiltration volume (UV), blood pressure, heart rate, and inferior caval vein (ICV) diameter. METHODS: In 10 patients on chronic haemodialysis, RBV measurement was performed during a total of one hundred 4-h haemodialysis sessions. Blood pressure and heart rate were measured at 5-min intervals. ICV diameter was assessed at the start and at the end of dialysis using ultrasonography. RESULTS: The changes in RBV showed considerable inter-individual variability. The average change in RBV ranged from 0.5 to -8.2% at 60 min and from -3.7 to -14.5% at 240 min (coefficient of variation (CV) 0.66 and 0.35 respectively). Intra-individual variability was also high (CV at 60 min 0.93; CV at 240 min 0.33). Inter-individual as well as intra individual variability showed only minor improvement when RBV was corrected for UV. We found a significant correlation between RBV and UV at 60 (r= -0.69; P<0.001) and at 240 min (r= -0.63; P<0.001). There was a significant correlation between RBV and heart rate (r= -0.39; P<0.001), but not between RBV or UV and blood pressure. The level of RBV reduction at which hypotension occurred was also highly variable. ICV diameter decreased from 10.3+/-1.7 mm/m(2) to 7.3+/-1. 5 mm/m(2). There was only a slight, although significant, correlation between ICV diameter and RBV (r= -0.23; P<0.05). The change in ICV-diameter showed a wide variation. CONCLUSIONS: RBV changes during haemodialysis showed a considerable intra- and inter-individual variability that could not be explained by differences in UV. No correlation was observed between UV or changes in RBV and either blood pressure or the incidence of hypotension. Heart rate, however, was significantly correlated with RBV. Moreover, IVC diameter was only poorly correlated with RBV, suggesting a redistribution of blood towards the central venous compartment. These data indicate that RBV monitoring is of limited use in the prevention of dialysis-related hypotension, and that the critical level of reduction in RBV at which hypotension occurs depends on cardiovascular defence mechanisms such as sympathetic drive. PMID- 10809810 TI - Gel clot LAL assay in the initial management of peritoneal dialysis patients with peritonitis: a retrospective study. AB - BACKGROUND: Indiscriminate use of broad-spectrum antibiotic treatment of peritonitis in peritoneal dialysis patients may have either unwanted side-effects or contribute to the development of antibiotic resistance. This may be avoided by improved diagnosis at presentation. The Limulus amoebocyte lysate assay is a convenient test detecting bacterial endotoxins or fungal beta glucans. This study evaluates a qualitative Limulus amoebocyte lysate test as a diagnostic tool used at presentation of a peritoneal dialysis patient with peritonitis. METHODS: One hundred and eleven episodes of peritonitis in peritoneal dialysis patients have been analysed retrospectively. Limulus amoebocyte lysate results at presentation were compared with culture results. A Limulus amoebocyte lysate assay was performed using a commercial kit by incubating a mixture of dialysate effluent and Limulus amoebocyte lysate reagent at 37 degrees C. The development of a stable solid clot was considered positive. The specificity and sensitivity of the test were calculated. RESULTS: The specificity of the Limulus amoebocyte lysate assay was found to be 98% and the sensitivity 74%. Limulus amoebocyte lysate assay was false-negative in 13 cases of Gram-negative peritonitis (22%). Limulus amoebocyte lysate was positive in three of seven cases of fungal peritonitis. The study included one case each with false-positive Limulus amoebocyte lysate and with culture-negative peritonitis. CONCLUSIONS: The Limulus amoebocyte lysate assay is a convenient and valuable diagnostic tool for excluding Gram-positive peritonitis in peritoneal dialysis patients. This allows more specific antibiotic treatment at presentation and may avoid the development of bacterial resistance. A negative Limulus amoebocyte lysate test is not reliable for the exclusion of Gram-negative peritonitis. In the absence of a positive culture result 48 h after presentation, accompanied by a delayed response to treatment, a positive Limulus amoebocyte lysate assay may indicate the presence of fungus. This justifies early empiric antifungal treatment before definitive culture results are made available. Routine Limulus amoebocyte lysate assay of dialysate effluent from continuous ambulatory peritoneal dialysis patients presenting with peritonitis is recommended. PMID- 10809811 TI - Effects of atorvastatin on dyslipidaemia in uraemic patients on peritoneal dialysis. AB - BACKGROUND: Our purpose was to evaluate the efficacy and safety of atorvastatin, a potent cholesterol- and triglyceride-lowering agent, in peritoneal dialysis patients with dyslipidaemia. METHODS: Peritoneal dialysis patients with hypercholesterolaemia were treated for 4 months with atorvastatin at a starting dose of 10 mg. The dose could be increased to 20 or 40 mg in order to achieve the following targets: plasma LDL-cholesterol of 130 mg/dl for primary prevention of coronary heart disease, plasma LDL cholesterol of 100 mg/dl for secondary prevention, and plasma triglycerides of 200 mg/dl. Plasma lipid profile and liver and muscle enzyme levels were assessed at baseline and then monthly during treatment. RESULTS: Thirty-one patients with hypercholesterolaemia were included (16 males and 15 females; mean age 57+/-16 years; mean duration of peritoneal dialysis 27+/-17 months). Nineteen of the patients also had hypertriglyceridaemia and seven had diabetes. Twenty patients had no coronary history (primary prevention), whereas nine had experienced a coronary event (secondary prevention). In the primary and the secondary prevention patients, mean LDL cholesterol levels (mg/dl) decreased significantly by 42 and 46% from 204+/-23 to 119+/-27 (P<0. 001) and 198+/-37 to 104+/-21 (P<0.001), and mean triglyceride levels (mg/dl) decreased by 37 and 26% from 289+/-132 to 186+/-92 (P<0.001) and 201+/-62 to 150+/-54 (P<0.001 respectively). Nineteen primary prevention and seven secondary prevention patients achieved the LDL-cholesterol target. The triglyceride target was achieved by 15 of the 19 hypertriglyceridaemic patients. Two patients stopped treatment (one because of gastrointestinal disturbances, the other because of an allergic skin reaction). After 4 months, there were no changes in enzyme levels. CONCLUSION: Atorvastatin is an effective and safe lipid lowering agent for peritoneal dialysis patients with mixed dyslipidaemia. PMID- 10809812 TI - A clinicopathological study of IgA nephropathy in renal transplant recipients: beneficial effect of angiotensin-converting enzyme inhibitor. AB - BACKGROUND: Prolonging the survival of transplant kidneys is a major task of modern nephrology. It has recently been shown that deteriorating renal function and substantial graft loss were observed in 55% of renal allograft recipients with recurrent IgA nephropathy (IgAN) at long-term follow-up. To gain a useful insight into the therapeutic approach towards protecting allograft kidneys from deteriorating graft function, we compared the histological characteristics of post-transplant IgAN to primary IgAN and investigated the effects of an ACE inhibitor. METHODS: Twenty-one patients with post-transplant IgAN and 63 patients with primary IgAN were included in the histopathological study. The effectiveness of angiotensin-converting enzyme (ACE) inhibitor treatment in post-transplant IgAN was also studied in 10 patients. RESULTS: The prevalence of glomeruli with adhesions and/or cellular crescents in primary IgAN was significantly greater than in post-transplant IgAN (P<0.05), but the proportion of glomeruli with segmental sclerosis was similar in both groups. The rate of global obsolescence, and the degree of interstitial fibrosis in post-transplant IgAN were significantly greater than in primary IgAN (P<0.05). The degree of glomerular obsolescence and the severity of interstitial fibrosis correlated with the severity of glomerular lesion in primary IgAN, but not in post-transplant IgAN. In primary IgAN, glomerular diameter significantly correlated with the proportions of glomerular obsolescence, but not in post-transplant IgAN, suggesting that allograft kidneys may be in a hyperfiltration state. Both the blood pressure and the urinary protein excretion significantly improved after ACE inhibitor treatment (P<0.001). CONCLUSION: In post-transplant IgAN, histopathological lesions indicative of acute inflammatory insults were suppressed, and glomerular hypertrophy, which may relate to haemodynamic burden such as hyperfiltration, was prominent. Preliminary study of ACE-inhibitor treatment in 10 patients showed favourable effects. A future long-term follow-up study is required to establish the effectiveness of ACE inhibitors in treatment of post-transplant IgAN. PMID- 10809813 TI - Age-matching in renal transplantation. AB - BACKGROUND: So far, the combined influence of donor age and recipient age on renal allograft survival has not been investigated sufficiently. In this retrospective single-centre study we analysed whether the influence of donor age and recipient age on renal allograft survival are dependent on each other. METHODS: Data from 1269 cadaveric renal allograft transplantations were evaluated. Paediatric donors (<15 years) and paediatric recipients (<15 years) were excluded. Donors and recipients were divided by age: young donors (yd, 55 years, n=176), young recipients (yr, 55 years, n=211). Functional and actual long-term graft survival (8 years) within the four resulting groups was determined: yd/yr (n=926), yd/or (n=167), od/yr (n=132), and od/or (n=44). RESULTS: Univariate analysis showed that long-term graft survival of both, kidneys from young donors (functional, 66.1 vs 52.2%, P=0.004; actual, 53.3 vs 46.2%, P=0.065) and kidneys from old donors (functional, 68.7 vs 22.5%, P=0.07; actual, 57.1 vs 20.8%, P=0.15) was better in old recipients as compared to young recipients. Multivariate regression analysis revealed that actual graft survival of kidneys from old donors was significantly reduced in young recipients (od/yr) as compared to all other groups (P=0.001; RR, 1. 97; 95% CI, 1.32-2.94). In this group of patients, graft loss was mainly due to acute (33.7%) and chronic (24.0%) rejection. CONCLUSION: Transplantation of kidneys from 'old' donors into 'young' recipients should be avoided, and these kidneys should be given to age-matched recipients. PMID- 10809814 TI - Survival of patients who have been on a waiting list for renal transplantation. AB - INTRODUCTION: Survival of transplanted patients is generally much better than for those on dialysis. This comparison is, however, incorrect, as in order to be accepted for renal transplantation the patient has to be in a relatively good condition and in addition transplanted patients are usually younger. We compared survival of all renal replacement therapy (RRT) patients who had undergone an identical medical check-up, been accepted, and put on the waiting list for cadaveric-kidney transplantation at Huddinge University hospital. A comparison with patients who were transplanted with a kidney from a living related donor (LD) is also included. METHODS: All patients (n=608) accepted and on the waiting list for renal transplantation between January 1987 and April 1996 formed the basis of the study. Follow-up was terminated on 31 December 1997. Survival was recorded from the date that the patients were accepted and put on the waiting list. As long as the patient was not transplanted and remained on dialysis treatment, survival was considered as 'survival on dialysis', and if transplanted, subsequent survival was defined as 'survival after cadaveric-kidney transplantation'. A patient who had been transplanted remained in that group for the rest of the observation period even if the transplantation had failed and the patient had to go back to dialysis after the surgery. RESULTS: Five-year survival was considerably better after LD-kidney transplantation (94%), than after cadaveric-kidney transplantation (76%) or on chronic dialysis (60%). Cox hazard regression analysis gave an age-adjusted relative risk for death of 0.46 for LD transplanted and 1.49 for remaining on dialysis compared with cadaveric transplanted patients. Transplanted patients, however, experienced a higher mortality during the first year after the transplantation than patients still on dialysis. CONCLUSIONS: LD-kidney transplantation is clearly associated with a superior survival. Mortality is relatively high after cadaveric-kidney transplantation, especially during the first months after surgery. Nevertheless, in the long term cadaveric kidney transplanted patients have a better survival than those remaining on dialysis. PMID- 10809815 TI - Adverse effect of donor arteriolosclerosis on graft outcome after renal transplantation. AB - INTRODUCTION: In recent years less strict criteria for renal graft donors have been applied. Our study was designed to investigate whether the histological picture, with special reference to vascular changes of the donor kidney, has an effect on the development and level of graft function, and on 48-month graft survival. METHODS: Three morphologically distinct groups were formed from 150 consecutive cadaveric kidneys donors transplanted into 290 recipients. A control group (C) consisted of kidneys with a completely normal histological picture. Group M1 included kidneys with mild arteriolosclerosis and group M2 (n=122) was comprised of kidneys showing significant arteriolosclerosis. The onset of graft function was assessed by the need for dialysis treatment post-transplantation and the levels of serum creatinine and creatinine clearance at 6, 12, 24 and 36 months post transplant. RESULTS: The proportion of sclerotic glomeruli (P<0.001) and the incidence and severity of interstitial fibrosis was greater in groups M1 and M2 than in the control group (M1, P<0.01; M2, P<0.001). The incidence of vascular fibrinoid necrosis in M2 was greater than in controls (P<0.001). The onset of graft function did not differ significantly between the groups. Group M2 showed a significantly lower level of graft function (P<0.001). The 4-year graft survival rate of group M2 was 74.2%, significantly lower than in the combined group C+M1 (P=0.03). CONCLUSION: Significant vascular lesions in the donor kidney should be taken into account when predicting graft function and survival. PMID- 10809816 TI - The influence of mycophenolate mofetil on the incidence and severity of primary cytomegalovirus infections and disease after renal transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) is increasingly used for immunosuppression after renal transplantation (RTx). The aim of our study was to investigate if the use of MMF has resulted in an increase in the frequency and severity of primary cytomegalovirus (CMV) infections. METHODS: Retrospective study of adult RTx patients who were CMV seronegative and who received a kidney of a CMV seropositive donor in the period 1992-1997 (n=84). Twenty-four of these patients were treated with MMF (in combination with cyclosporin and prednisone; MMF+) and the other 60 were the control group (cyclosporin and prednisone; MMF-). No CMV prophylaxis was given. CMV infection was defined as CMV seroconversion of IgG antibodies. CMV disease was defined as CMV infection and fever in combination with one or more of the following: leukocytopenia, thrombocytopenia, elevated alanine aminotransferase, or histological evidence of tissue invasive disease. RESULTS: The incidence of primary CMV infections was similar in both groups (MMF+, 75%; MMF-, 63%). CMV disease was more frequent in the MMF+ group than in the MMF- group (67 vs 30%, P<0.05). In the patients with CMV disease, the use of MMF did not affect severity of symptoms, frequency of tissue invasive disease, or frequency or duration of treatment with ganciclovir. CONCLUSIONS: Addition of MMF to the immunosuppressive therapy after RTx did not result in an increase of primary CMV infections.However, these CMV infections led more often to CMV disease in patients treated with MMF than in those without MMF. PMID- 10809817 TI - Baclofen neurotoxicity [correction of unerotoxicity] in a chronic haemodialysis patient. PMID- 10809818 TI - Concurrent pheochromocytoma, diabetes insipidus and cerebral venous thrombosis--a possible unique pathophysiological mechanism. PMID- 10809819 TI - Unsuspected cardiac tamponade following insertion of a haemodialysis catheter: a normal chest radiograph does not exclude a complication. PMID- 10809820 TI - Implantation of a new device for haemodialysis. PMID- 10809821 TI - Green urine after motorcycle accident. PMID- 10809822 TI - The patient who had a picnic at a waterfall and presented with haemoptysis and renal failure. PMID- 10809823 TI - Beware backache in chronic dialysis patients. PMID- 10809825 TI - Berndt tersteegen award 2000 PMID- 10809824 TI - Pulmonary, cardiac and renal syndrome. PMID- 10809826 TI - Unexplained haematuria. PMID- 10809827 TI - Clearance of an oral delayed-release mesalazine preparation (Salofalk) by haemodialysis--pharmacokinetic profile of mesalazine related to the effects of dialysis. PMID- 10809829 TI - Clearance of an oral delayed-release mesalazine preparation (Salofalk((R))) by haemodialysis-pharmacokinetic profile of mesalazine related to the effects of dialysis PMID- 10809828 TI - Can antihypertensive medications control BP in haemodialysis patients. PMID- 10809831 TI - Renal terminology. PMID- 10809830 TI - Long-term efficacy of lipoprotein apheresis in homozygous familial hypercholesterolaemia. PMID- 10809832 TI - Long-term efficacy of lipoprotein apheresis in homozygous familial hypercholesterolaemia PMID- 10809833 TI - Influenza vaccine--a possible trigger of rhabdomyolysis induced acute renal failure due to the combined use of cerivastatin and bezafibrate. PMID- 10809834 TI - Acute renal failure after transarterial chemoembolization progressing to chronic renal failure in hepatocellular carcinoma. PMID- 10809835 TI - The beneficial effect of a journalist's death on organ transplantation and hepatitis B vaccination. PMID- 10809836 TI - Isolated tuberculous pyomyositis in a renal transplant patient. PMID- 10809837 TI - Announcements PMID- 10809838 TI - Cutaneous CD56 natural killer and natural killer-like T-cell lymphoma. PMID- 10809839 TI - Phototherapy and acne vulgaris. PMID- 10809840 TI - The usage of complementary therapies by dermatological patients: a systematic review. AB - Complementary medicine (CM) is more popular than ever before. Dermatology has not remained unaffected by this trend. The aim of this systematic review was to summarize all surveys of dermatological patients regarding the usage of CM. Three independent literature searches were carried out. Data were extracted in a predefined, standardized way and evaluated descriptively. Seven surveys met the inclusion/exclusion criteria. Collectively they show a high but variable prevalence of CM. Lifetime prevalence ranged from 35 to 69%. The most frequently used treatment modalities comprise homoeopathy, herbalism and food supplements. With this high level of prevalence, research into the potential risks and benefits of CM is urgently needed. Dermatologists should consider discussing CM openly with their patients. PMID- 10809841 TI - The human hair follicle immune system: cellular composition and immune privilege. AB - The immunology of the hair follicle, its relationship with the 'skin immune system' and its role in hair diseases remain biologically intriguing and clinically important. In this study, we analysed the immunoreactivity patterns of 15 immunodermatological markers to determine the cellular composition and immune privilege of the human hair follicle immune system in anagen VI (growth phase). The most prominent cells located in or around the hair follicle were Langerhans cells, CD4+ or CD8+ T cells, macrophages and mast cells, whereas B cells, natural killer cells and gammadelta T cells were found very rarely. Langerhans cells (CD1a+, major histocompatibility complex, MHC class II+), and T cells (CD4+ or CD8+) were predominantly distributed in the distal hair follicle epithelium, whereas macrophages (CD68+, MHC class II+) and mast cells (Giemsa+) were located in the perifollicular connective tissue sheath. Transmission electron microscopy confirmed low numbers of immune cells in the proximal hair follicle epithelium, and very few macrophages and Langerhans cells were seen in the dermal papilla. Melanophages were observed in the connective tissue sheath and dermal papilla. MHC class I (HLA-A, -B, -C) and beta2-microglobulin immunoreactivity was found on most skin cells, but was substantially reduced on isthmus keratinocytes and virtually absent in the proximal hair follicle epithelium. Apart from the absence of Fas ligand immunoreactivity, the sharply reduced numbers of T cells and Langerhans cells, and the virtual absence of MHC class I expression all suggest that the anagen proximal hair follicle constitutes an area of immune privilege within the hair follicle immune system, whose collapse may be crucial for the pathogenesis of alopecia areata. PMID- 10809842 TI - Modulation of skin collagen metabolism by irradiation: collagen synthesis is increased in irradiated human skin. AB - Radiation-induced fibrosis is a common side-effect of cancer treatment. The pathophysiological events leading to fibrosis are not known in detail. We analysed the effect of therapeutic irradiation on human skin collagen synthesis, skin thickness, gelatinases and their inhibitors. Twenty randomly chosen women who had been treated for breast cancer with surgery and radiation therapy participated in the study. In each patient, the irradiated skin area was compared with a corresponding non-treated skin area. Suction blister fluid (SBF) and serum samples were analysed for the aminoterminal propeptides of type I and type III procollagens (PINP and PIIINP), tissue inhibitors of matrix metalloproteinases (MMPs) 1 and 2 (TIMP-1 and TIMP-2) and MMP-9 and MMP-2/TIMP-2 complex. Skin biopsies were analysed for PINP and immunohistochemical staining was used for PIIINP. In irradiated skin, PINP, PIIINP, TIMP-1 and MMP-2/TIMP-2 complex levels in SBF and the number of PINP-positive fibroblasts in tissue sections were significantly higher in comparison with non-treated skin. The levels of TIMP-2 in irradiated and non-irradiated skin were similar. MMP-9 could not be detected in SBF with the assay used. The serum levels of MMP-9 were higher in the treated subjects than the reference values. The serum values of PINP, PIIINP, TIMP-1, TIMP-2 and MMP-2/TIMP-2 complex were not significantly affected. These results indicate increased local collagen synthesis and accumulation of connective tissue in irradiated skin. The marked upregulation of collagen synthesis as a result of irradiation offers a possibility to treat this complication with compounds such as topical steroids which downregulate collagen synthesis. PMID- 10809843 TI - Barrier recovery rate varies time-dependently in human skin. AB - The recovery in cutaneous barrier functions, assessed in terms of transepidermal water loss, 1 h after tape stripping of volar forearm skin in human volunteers, was investigated at different times over the 24 h day. The barrier recovery rate was significantly lower between 20:00 h and 23:00 h than that at other time points. The skin surface temperature and the basal transepidermal water loss reached their highest values at about 03:00 h (33.6 degrees C and 0.30 mg cm-2 h 1), while the cortisol level in the saliva was highest at 09:00 h (7.8 pmol mL 1). These results suggest significant time-dependent variation in cutaneous barrier repair independent of changes in skin temperature and cortisol level. PMID- 10809844 TI - The cutaneous microflora of adolescent, persistent and late-onset acne patients does not differ. AB - The cutaneous microbiology and antibody status to Propionibacterium acnes of patients with persistent (males, n = 32; females, n = 33) and late-onset (females, n = 25) acne were compared with individuals with adolescent acne (males, n = 22; females, n = 18) and normal control volunteers (persistent acne: males, n = 26; females, n = 30; late-onset: females, n = 20). Males had significantly higher grades of acne compared with females (P < 0.05). The microflora consisted in the main of propionibacteria, staphylococci and Malassezia; other bacteria represented less than 0.01% of the total microflora. At all sites for all samples there were significantly more propionibacteria than staphylococci or Malassezia (P < 0.05). There were significantly higher (P < 0.05) numbers of microorganisms in follicular casts from patients compared with their control volunteers for female facial skin and male back skin. Twenty-six papules and 48 normal follicles were analysed. A bimodal distribution of microbial colonization was noted, with about 90% of normal follicles and about 10% of acne follicles having no detectable viable microorganisms. Anti-P. acnes IgG antibody titres were measured using a secondary fluorescein isothiocyanate antibody technique, and no significant differences in titre were found between any groups of patients (P > 0.05). Correlation analysis showed no association between the population densities of P. acnes and anti-P. acnes IgG titres. There were no differences in the microbiology of skin of adolescent acne patients, persistent acne patients or late-onset acne patients which could account for these various forms of acne. PMID- 10809845 TI - Sensitivity and specificity of epiluminescence microscopy: evaluation on a sample of 2731 excised cutaneous pigmented lesions. The Melanoma Cooperative Study. AB - To evaluate the role of epiluminescence microscopy (ELM) in the differential diagnosis of cutaneous pigmented lesions, and to improve the early diagnosis of cutaneous malignant melanoma (CMM), 15,719 pigmented lesions from 8782 consecutive patients were evaluated using ELM with a hand-held video microscope imaging system (MS 500B Micro-Scopeman, Moritex). Comparison between risk levels as inferred from ELM screening and histology was performed on 2731 surgically excised lesions. ELM sensitivity, specificity, positive and negative predictive values, as well as agreement with histological results for the different subgroups of lesions, were determined. Overall agreement was 87.3% (ranging from 85.1% to 92.2% for melanocytic and non-melanocytic lesions, respectively); sensitivity and specificity were high (values ranging from 87.3% to 96.3% among different subsets of ELM-analysed lesions) and statistically significant (P < 0.0001). ELM screening identified 165 new cases of CMM with a high proportion of lesions (115; 70%) in an early phase of tumour growth (Breslow thickness 50%) than in MALT- or FCC-type lymphomas (< 10%). Features associated with an unfavourable prognosis were the presence of multiple skin lesions at diagnosis, transformation from MALT-type lymphoma to DLBCL, and possibly p16INK4a aberrations. It is concluded that most CBCLs are dissimilar from FCC lymphomas and seem to be more closely related to marginal zone/MALT-type lymphomas. It is also suggested that there are fundamental differences between DLBCL and other histological categories of CBCL, indicating that cutaneous DLBCL is a separate entity with an increased growth potential and genetic features similar to DLBCL originating in other anatomical sites. PMID- 10809849 TI - Clinicopathological features of CD56+ nasal-type T/natural killer cell lymphomas with lobular panniculitis. AB - Nasal-type T/natural killer cell lymphoma (TNKCL) shows frequent extranodal involvement including the skin, and is associated with a poor prognosis. We have studied six patients with nasal-type TNKCL presenting with inflammatory subcutaneous nodular lesions with a subcutaneous lymphoid infiltrate. Clinical information was obtained from the medical records of the patients and at follow up. All cases showed features of angiocentric lymphoma on histology, although there was diffuse cellular infiltration rather than an angiocentric pattern in the subcutis. All six patients were similar in immunophenotype: positive for CD56 and either cytoplasmic CD3 or CD45RO, but negative for B-cell markers and CD30. In situ hybridization using an anti-sense Epstein-Barr virus early regions probe showed a positive reaction in all cases. All patients either died with progressive disease or showed no response to combined chemotherapy. The diagnosis of nasal-type TNKCL, which has a fatal outcome, is facilitated by detection of CD56-positive tumour cells. In evaluating lobular panniculitis including apparently benign inflammatory subcutaneous nodules, nasal-type TNKCL should be considered in the differential diagnosis, especially in Asian countries. PMID- 10809850 TI - Topical corticosteroid phobia in patients with atopic eczema. AB - Topical corticosteroids are widely prescribed by dermatologists caring for patients with atopic eczema. Patients' fears about using topical corticosteroids may have important implications for compliance with treatment. We carried out a questionnaire-based study of 200 dermatology outpatients with atopic eczema (age range 4 months-67.8 years) to assess the prevalence and source of topical corticosteroid phobia. We also questioned patients on their knowledge of the potencies of different topical corticosteroids. Overall, 72.5% of people worried about using topical corticosteroids on their own or their child's skin. Twenty four per cent of people admitted to having been non-compliant with topical corticosteroid treatment because of these worries. The most frequent cause for concern was the perceived risk of skin thinning (34.5%). In addition, 9.5% of patients worried about systemic absorption leading to effects on growth and development. The most commonly used topical corticosteroid was hydrocortisone, yet 31% of patients who used this preparation classified it as either strong, very strong or did not know the potency. Only 62.5% of the 48 patients who had used both Dermovate (Glaxo) and hydrocortisone in the past were able to correctly grade Dermovate as being more potent than hydrocortisone. The most common source of patient information regarding topical corticosteroid safety was the general practitioner. Although skin thinning and systemic effects can develop very occasionally in people using topical corticosteroids, the concern expressed by people using them seems out of proportion in relation to the evidence of harm. This study highlights the need for provision of better information and education to patients and possibly general practitioners regarding the safety, potency and appropriate use of topical corticosteroids. PMID- 10809851 TI - Clinical features of 31 patients with systemic contact dermatitis due to the ingestion of Rhus (lacquer). AB - In Korea, Rhus has been used as a folk medicine to cure gastrointestinal diseases and as a health food. We review the clinicopathological and laboratory findings in patients with systemic contact dermatitis caused by intake of Rhus. We reviewed medical records and histopathological sections from 31 patients during a 10-year period. The male/female ratio was 1.4: 1 and the average age was 43.8 years (range 22-70). Ten patients (32%) had a known history of allergy to lacquer. Rhus was ingested to treat gastrointestinal problems including indigestion and gastritis (45%), and as a health food (39%), in cooked meat, in herbal medicine, or taken by inhalation. The patients developed skin lesions such as a maculopapular eruption (65%), erythema multiforme (EM, 32%), erythroderma (19%), pustules, purpura, weals and blisters. Erythroderma was very frequent in patients with a known history of allergy to lacquer, but maculopapular and EM type eruptions were more frequently observed in those without a history of allergy. All patients experienced generalized or localized pruritus. Other symptoms included gastrointestinal problems (32%), fever (26%), chills and headache; many developed leucocytosis (70%) with neutrophilia (88%), while some showed toxic effects on liver and kidney. Fifty-nine per cent of patients observed cutaneous or general symptoms within a day after ingestion of Rhus. There was no difference in the time lag for symptoms to develop between patients allergic and not allergic to Rhus. All patients responded well to treatment with systemic steroids and antihistamines. Common histopathological findings were vascular dilatation, perivascular lymphohistiocytic infiltration, and extravasation of red blood cells in the upper dermis. Rhus lacquer should not be ingested in view of its highly allergic and toxic effects. PMID- 10809852 TI - Microbiology of infected poison ivy dermatitis. AB - We report the aerobic and anaerobic microbiology of secondarily infected poison ivy dermatitis. The study involved retrospective review of clinical and microbiology laboratory records of patients with secondarily infected poison ivy lesions. Bacterial growth was noted in 33 specimens. Aerobic or facultative anaerobic bacteria only were present in 18 (55%) patients, anaerobic bacteria only in seven (21%), and mixed anaerobic-aerobic bacteria in eight (24%). Forty five isolates were recovered (1.4 per specimen): 27 aerobic or facultative anaerobic bacteria, and 18 strict anaerobes. The predominant aerobic and facultative anaerobic bacteria were Staphylococcus aureus (13 isolates) and group A beta-haemolytic streptococci (six). The predominant anaerobes were Peptostreptococcus spp. (seven isolates), pigmented Prevotella and Porphyromonas spp. (four) and Fusobacterium spp. (two). Single bacterial isolates were recovered in 18 (55%) patients, eight of which were S. aureus. Nineteen of the organisms isolated from 16 (48%) patients produced the enzyme beta-lactamase. Organisms that resided in the mucous membranes close to the lesions predominated in those infections. Enteric gram-negative rods and Bacteroides fragilis group predominated in leg and buttock lesions. Group A beta-haemolytic streptococci, pigmented Prevotella and Porphyromonas and Fusobacterium spp. were most frequently recovered from lesions of the finger, face and neck. The polymicrobial aetiology of secondarily infected poison ivy lesions, and the association of bacterial flora with the anatomical site of the lesions, are demonstrated. PMID- 10809853 TI - The clinical genetics of hidradenitis suppurativa revisited. AB - A familial form of hidradenitis suppurativa (HS) with autosomal dominant inheritance was described in a study conducted 15 years ago in Nottingham but has not been systematically confirmed elsewhere. Prior to commencing molecular genetic studies, we wanted to test the validity of the previous study by assessing its reproducibility on the basis of a strict, newly devised disease definition for HS. We were also interested whether new cases of the disease had arisen meantime in the study group as should be expected for an autosomal dominant disease. We reviewed 14 surviving probands and their families. Seven of these probands had previously been noted to have a positive family history whereas the others had been classified as having a negative or possible family history. One hundred and thirty-two family members were assessed for their respective disease status. Participants were initially contacted by telephone or letter, and those who acknowledged a history of at least one previous boil were invited for a personal examination and interview. Only personally examined individuals were classified as a case. Twenty-eight relatives with HS were detected in total, and 27 of these were in the group previously labelled family history positive. Nine of these cases had not been detected in the previous study and in at least seven of these the disease had developed after the previous study had been conducted. Only twice did our criteria fail to confirm cases that had been labelled as HS in the previous study. Both times we classified the patients as 'possibly affected'. A further 16 relatives were judged to be possibly affected. In the group with positive family history we found 10 affected and nine possibly affected individuals among 37 surviving first-degree relatives of HS sufferers. Our findings support the concept of a familial form of HS with autosomal dominant inheritance. An insufficiently sensitive disease definition, a variable degree of gene penetrance and possibly a hormonal influence on gene expression may explain the reduced risk to first-degree relatives, which falls short of the expected 50% mark. Molecular genetic studies to clarify whether one or more gene(s) are involved in HS are now necessary and have been commenced. PMID- 10809854 TI - Schnitzler syndrome: heterogeneous immunopathological findings involving IgM-skin interactions. AB - The Schnitzler syndrome is the association of chronic urticaria, intermittent fever, osteosclerotic bone lesions and a monoclonal IgM gammopathy. It is not yet firmly established whether the monoclonal immunoglobulin component plays a part in the pathophysiology of the urticarial lesions. Immunoblotting on epidermal and dermal human skin extracts as well as immunoelectron microscopic (IEM) studies on Lowicryl K4M-embedded skin sections were performed in three patients with the Schnitzler syndrome. Western blotting on epidermal extracts showed the presence of IgM-kappa antiskin autoantibodies in two patients. These antibodies displayed the same isotype as the monoclonal components and recognized a 280-290-kDa antigen in one patient and a 200-kDa antigen in the other patient. IEM studies showed sparse IgM deposits in the epidermis, around the keratinocytes, near the desmosomes in one patient and dense deposits below the lamina densa, in the region of the anchoring fibrils, in another patient. Antiskin IgM autoantibodies of the same isotype as their monoclonal gammopathies can be present in the serum of some patients with the Schnitzler syndrome. These IgM antibodies seem to deposit in vivo in the epidermis and at the dermal-epidermal junction, in the region of the anchoring fibrils. These findings suggest that the monoclonal gammopathy plays a part in the pathophysiology of the skin rash. They also suggest patient heterogeneity both in the skin antigens that are recognized as well as in their localization. PMID- 10809855 TI - A multicentre study of percentage change in venous leg ulcer area as a prognostic index of healing at 24 weeks. AB - Venous leg ulcers, which may take months to heal, account for 40-70% of all lower extremity chronic wounds. New treatment options for venous leg ulcers have recently been proposed, and therefore deciding which patients are candidates for these novel-and often expensive-treatments is an important task. Moreover, researchers conducting clinical trials often wish to enroll patients who are unlikely to heal in order to minimize sample sizes needed and research costs. Our purpose was to assess the use of percentage change in venous leg ulcer area over the first few weeks of treatment as a prognostic indicator of healing or non healing at 24 weeks. We conducted a cohort study based on an existing data set from a multicentre randomized clinical trial that enrolled 104 patients. Wounds were measured using digital planimetry for 4 consecutive weeks following the inception of good wound care. Utilizing the Wilcoxon rank sum (Mann-Whitney) test, we found that percentage change in area over time distinguished between those who healed and those who failed to heal after 24 weeks of good wound care (P < 0.05). The rate of healing, or area healed per week, did not differentiate between those who healed at 24 weeks and those who did not, as all patients had similar rates of healing over the first 4 weeks of treatment. Percentage change in area from baseline to week 4 provided the best combination of positive and negative predictive values (68.2%, 74.7%) and the largest area under the receiver operating characteristic curve (0.75). Thus, percentage change in area over the first 4 weeks of treatment represents a practical and predictive measure of complete wound healing by 24 weeks. PMID- 10809856 TI - Once weekly fluconazole is effective in children in the treatment of tinea capitis: a prospective, multicentre study. AB - In an open, multicentre evaluation carried out in Brazil, Canada and South Africa we have demonstrated that fluconazole 8 mg kg-1 once weekly is effective in tinea capitis caused by Trichophyton and Microsporum species. There were 61 children, aged (mean +/- SE) 5.0 +/- 0.3 years; weight (mean +/- 5.6) 20.0 +/- 0.9 kg; 41 males, 20 females; one Asian, 57 Black, one Caucasian and two Hispanic. The organisms were Trichophyton violaceum (33 patients), T. tonsurans (11) and Microsporum canis (17). The extent of tinea capitis at pretherapy was: mild (18 patients), moderate (30) and severe (13). Patients with tinea capitis due to Trichophyton species were initially treated for 8 weeks with an extra 4 weeks of fluconazole if clinically indicated. All 44 patients with tinea capitis due to Trichophyton species were completely cured (clinically and mycologically) when evaluated 8 weeks after completion of active treatment, following 8 weeks of once weekly dosing in 35 patients and 12 weeks of once weekly dosing in nine patients. In Microsporum canis tinea capitis, an extra 4 weeks was administered at week 12 in patients where it was clinically indicated at the time. Sixteen of 17 patients with M. canis tinea capitis were completely cured (clinically and mycologically) when evaluated 8 weeks following the end of treatment when given for 8, 12 and 16 weeks in 12, one and three patients, respectively. Overall, complete cure (clinical and mycological) occurred in 60 of 61 patients at follow-up 8 weeks from the end of therapy. The duration of once weekly fluconazole in the 60 patients was 8 weeks (47 patients), 12 weeks (10 patients) and 16 weeks (three patients), respectively. Clinical adverse effects consisted of a mild, reversible gastrointestinal complaint in three (4.9%) of 61 children. A laboratory abnormality with elevated liver function tests was observed in one (5.9%) of 17 patients; this was asymptomatic, and reversible. No patient discontinued therapy. The data suggest that once weekly fluconazole dosing is effective, safe and associated with high compliance when used to treat tinea capitis. PMID- 10809857 TI - Ivermectin alone or in combination with benzyl benzoate in the treatment of human immunodeficiency virus-associated scabies. AB - In order to establish a safe and reliable treatment for human immunodeficiency virus (HIV)-associated scabies, we have treated 60 episodes of scabies in this setting, occurring in 39 patients, with one of the following regimens: (i) topical treatment with benzyl benzoate solution; (ii) single-dose oral treatment with ivermectin alone; and (iii) combination therapy with benzyl benzoate solution and oral ivermectin, employing the same regimens as single-agent therapy. Patients were stratified according to the severity score of the disease and the outcome (eradication, relapse, failure). We found that both benzyl benzoate and ivermectin alone were quite effective in mild to moderate scabies, but they were both associated with an unacceptable rate of relapse and failure in severe or crusted scabies. In contrast, combined treatment produced an optimal rate of success, without significant treatment-related side-effects. Therefore, we consider that combination treatment with benzyl benzoate solution and oral ivermectin is preferable to single-agent therapy in crusted scabies occurring in HIV/acquired immune deficiency syndrome patients. PMID- 10809858 TI - Phototherapy with blue (415 nm) and red (660 nm) light in the treatment of acne vulgaris. AB - In this study we have evaluated the use of blue light (peak at 415 nm) and a mixed blue and red light (peaks at 415 and 660 nm) in the treatment of acne vulgaris. One hundred and seven patients with mild to moderate acne vulgaris were randomized into four treatment groups: blue light, mixed blue and red light, cool white light and 5% benzoyl peroxide cream. Subjects in the phototherapy groups used portable light sources and irradiation was carried out daily for 15 min. Comparative assessment between the three light sources was made in an observer blinded fashion, but this could not be achieved for the use of benzoyl peroxide. Assessments were performed every 4 weeks. After 12 weeks of active treatment a mean improvement of 76% (95% confidence interval 66-87) in inflammatory lesions was achieved by the combined blue-red light phototherapy; this was significantly superior to that achieved by blue light (at weeks 4 and 8 but not week 12), benzoyl peroxide (at weeks 8 and 12) or white light (at each assessment). The final mean improvement in comedones by using blue-red light was 58% (95% confidence interval 45-71), again better than that achieved by the other active treatments used, although the differences did not reach significant levels. We have found that phototherapy with mixed blue-red light, probably by combining antibacterial and anti-inflammatory action, is an effective means of treating acne vulgaris of mild to moderate severity, with no significant short-term adverse effects. PMID- 10809859 TI - Topical application of St John's wort (Hypericum perforatum L.) and of its metabolite hyperforin inhibits the allostimulatory capacity of epidermal cells. AB - St John's wort (Hypericum perforatum) is a traditional herbal medicine that is used for the topical treatment of superficial wounds, burns and dermatitis. The characteristic metabolites of St John's wort are the photodynamic active plant pigment hypericin and the phloroglucin-derivative hyperforin. To date, no studies on immunomodulatory properties of topical preparations of St John's wort have been performed. Here, we investigated the alloantigen presenting function of human epidermal cells (EC) exposed to Hypericum ointment in vivo in a mixed EC lymphocyte reaction (MECLR). The effect of Hypericum ointment was compared with the immunosuppressive effect of solar-simulated radiation (SSR). Subsequently, we tested purified hyperforin in vivo and in vitro in a MECLR to evaluate its possible contribution to the effect of the Hypericum ointment. Furthermore, we assessed the effect of hyperforin on the proliferation of peripheral blood mononuclear cells (PBMC) in vitro. Compared with untreated skin, treatment with Hypericum ointment resulted in a significant suppression of the MECLR (P 500 mg l 1, arterial pressure: systolic > 140 mmHg, diastolic > 100 mmHg). Radioimmunological techniques were employed to determine IGF-I and IGF-BPs (BP-1 and BP-3). RESULTS: It was found that pre-eclampsia is associated with an increase of IGF-I concentration in the UC serum and with simultaneous decrease of its content in the umbilical cord artery (UCA). The decrease of IGF-I content in the UCA wall was accompanied by an increase of BP-3 and BP-1 in this tissue. The increase in BPs content in the UCA wall was not associated with an enhancement of IGF binding by extracts from the homogenates of arterial wall. Heparin drastically decreased the binding of IGF-I by BP-3. CONCLUSIONS: Pre-eclampsia is associated with an increase of IGF-I-concentration in the umbilical cord blood and an elevation of BPs contents in the UCA wall. Despite a high concentration of binding proteins, IGF-I is not accumulated in this tissue. High amounts of sulphated GAGs in the UCA wall may be a factor that prevents the binding of IGF-I by BPs. Free IGF-I can easily bind to cell receptors and stimulate the cells to produce collagen and sulphated GAGs in the arterial wall. PMID- 10809900 TI - Effect of hemipancreatectomy and of pancreatic diversion on the tolerance to a glucose load in humans. AB - BACKGROUND: The role of the pattern, quantity and site of insulin secretion in the tolerance to a glucose challenge is not fully evaluated in humans because it is difficult to obtain appropriate clinical models. DESIGN: To address this issue, we studied subjects with reduced pancreatic mass (hemipancreatectomized, HEMI), systemic insulin delivery (pancreas transplant recipients, PTX), and two control groups (healthy, CON; and with uveitis on the same immunosuppression as PTX, UVE), with an hyperglycaemic clamp (study 1, + 4.2 mmol L-1), using a repeat experiment (study 2) with a fixed glucose infusion, calculated to increase by 35% that in study 1. RESULTS: In study 1, CON increased glucose uptake to 20 +/- 3 micromol kg-1 min-1 after a biphasic insulin response. In study 2, CON further increased the glucose uptake via an increment in prehepatic insulin secretion that stimulated insulin sensitivity without changes in peripheral insulin and glucose concentrations. HEMI and PTX had 35% less glucose uptake in study 1, compared to CON, and increased glucose concentrations (+ 1.6 mmol L-1) in study 2. UVE had an intermediate defect. The causes of intolerance were different: HEMI had a defective first-phase insulin secretion (50% peripheral insulin concentrations) but maintained insulin sensitivity; PTX had normal peripheral insulin but only one-third of the insulin sensitivity of CON. CONCLUSIONS: Hemipancreatectomy and systemic insulin delivery impair first-phase insulin secretion; second-phase peripheral insulinization (HEMI); insulin sensitivity (PTX); and a mechanism evidentiated in study 2 of CON that increases insulin sensitivity in response to prehepatic insulin secretion (both groups). Failure of these mechanisms is largely compensated by hyperglycaemia. PMID- 10809901 TI - A novel hyperglycaemic clamp for characterization of islet function in humans: assessment of three different secretagogues, maximal insulin response and reproducibility. AB - BACKGROUND: Characterization of beta-cell function in humans is essential for identifying genetic defects involved in abnormal insulin secretion and the pathogenesis of type 2 diabetes. MATERIALS AND METHODS: We designed a novel test assessing plasma insulin and C-peptide in response to 3 different secretagogues. Seven lean, healthy volunteers twice underwent a 200 min hyperglycaemic clamp (10 mmol L-1) with administration of GLP-1 (1.5 pmol. kg-1. min-1) starting at 120 min and an arginine bolus at 180 min. We determined glucose-induced first and second-phase insulin secretion, GLP-1-stimulated insulin secretion, arginine stimulated insulin response (increase above prestimulus, DeltaIarg) and the maximal, i. e. highest absolute, insulin concentration (Imax). Insulin sensitivity was assessed during second-phase hyperglycaemia. On a third occasion 6 subjects additionally received an arginine bolus at > 25 mM blood glucose, a test hitherto claimed to provoke maximal insulin secretion. RESULTS: Insulin levels increased from 46 +/- 11 pM to 566 +/- 202 pM at 120 min, to 5104 +/- 1179 pM at 180 min and to maximally 8361 +/- 1368 pM after arginine (all P < 0.001). The within subject coefficients of variation of the different secretion parameters ranged from 10 +/- 3% to 16 +/- 6%. Except for second-phase which failed to correlate significantly with DeltaIarg (r = 0.52, P = 0.23) and Imax (r = 0.75, P = 0.053) all phases of insulin secretion correlated with one another. The insulin concentration after the arginine bolus at > 25 mM glucose (n = 6) was 2773 +/- 855 pM vs. 7562 +/- 1168 pM for Imax (P = 0.003). CONCLUSION: This novel insulin secretion test elicits a distinct pattern of plasma insulin concentrations in response to the secretagogues glucose, GLP-1 and arginine and is highly reproducible and can be used for differential characterization of islet function. PMID- 10809902 TI - Effects of exogenous zinc supplementation on intestinal epithelial repair in vitro. AB - BACKGROUND: Substitution of zinc modulates antioxidant capabilities within the intestinal mucosa and improves intestinal wound healing in zinc-deficient patients with inflammatory bowel diseases. The aim of this study was to characterize the modulating effects of zinc on intestinal epithelial cell function in vitro. MATERIALS AND METHODS: The effects of zinc on intestinal epithelial cell morphology were assessed by phase contrast and transmission electron microscopy using the non-transformed small intestinal epithelial cell line IEC-6. Zinc-induced apoptosis was assessed by DNA fragmentation analysis, lactate dehydrogluase (LDH) release and flow cytometry with propidium iodine staining. Furthermore, the effects of zinc on IEC-6 cell proliferation were assessed using a colorimetric thiazolyl blue (MTT) assay and on IEC-6 cell restitution using an in vitro wounding model. RESULTS: Physiological concentrations of zinc (25 microM) did not significantly alter the morphological appearance of IEC-6 cells. However, a 10-fold higher dose of zinc (250 microM) induced epithelial cell rounding, loss of adherence and apoptotic characteristics. While physiological zinc concentrations (< 100 microM) did not induce apoptosis, supraphysiological zinc concentrations (> 100 microM) caused apoptosis. Physiological concentrations of zinc (6.25-50 microM) had no significant effect on intestinal epithelial cell proliferation. In contrast, physiological concentrations of zinc (12.5-50 microM) significantly enhanced epithelial cell restitution through a transforming growth factor-beta (TGFbeta) independent mechanism. Simultaneous addition of TGFbeta and zinc resulted in an additive stimulation of IEC-6 cell restitution. CONCLUSION: Zinc may promote intestinal epithelial wound healing by enhancement of epithelial cell restitution, the initial step of epithelial wound healing. Zinc supplementation may improve epithelial repair; however, excessive amounts of zinc may cause tissue injury and impair epithelial wound healing. PMID- 10809903 TI - Production and action of transforming growth factor-beta in human osteoblast cultures: dependence on cell differentiation and modulation by calcitriol. AB - BACKGROUND: Transforming growth factor beta (TGF-beta) plays an important role in skeletal remodelling. However, few studies have examined its effects on cultured human osteoblasts. Our aim is to characterise the biological effects of TGF-beta1 on human osteoblasts and to examine the interaction between TGF-beta1 and calcitriol. DESIGN: In vitro study employing two models of normal human osteoblasts: human bone marrow stromal cells [hMS/(OB)] containing osteoprogenitor cells and trabecular bone osteoblasts (hOB), which are mature osteoblasts. A reverse-transcriptase-polymerase-chain-reaction assay was employed to measure steady state mRNA levels of TGF-beta(s) isoforms and receptors. Effects of short-term treatment of TGF-beta1 on osteoblast proliferation and differentiation markers were assessed. The effect of cotreatment of calcitriol (10-8 M) and TGF-beta1 on osteoblast differentiation was also determined. RESULTS: Both hMS(OB) and hOB cells expressed mRNA transcripts of TGF-beta1, TGF beta2, TGF-beta 3, TGF-beta type I and type II receptors. TGF-beta 1 stimulated osteoblast proliferation in hMS(OB) and in hOB cultures. In hOB cultures, TGF beta1 stimulated AP production and cotreatment with calcitriol induced a synergistic increase in AP levels to 250 +/- 61% of calcitriol-treated controls. Effects of TGF-beta1 and calcitriol were less pronounced in hMS(OB) cultures. TGF beta1 inhibited collagen type I production in hMS(OB) cells and these effects were abolished in presence of calcitriol. In presence of calcitriol, TGF-beta1 increased collagen type I production in hOB cells. In both hOB and hMS(OB) cultures, TGF-beta1 inhibited osteocalcin production. CONCLUSIONS: TGF-beta increases osteoblastic cell proliferation irrespective of the differentiation state. In presence of calcitriol, it initiates osteoblast cell differentiation and matrix formation. As TGF-beta inhibits osteocalcin production, other factors are necessary for inducing terminal differentiation of osteoblasts. The observed effects of TGF-beta on human osteoblasts in vitro may represent important regulatory steps in controlling osteoblast cell proliferation and differentiation in vivo. PMID- 10809904 TI - rhIGF-I/IGFBP-3 complex, but not free rhIGF-I, supports muscle protein biosynthesis in rats during semistarvation. AB - BACKGROUND: The aim of this study was to evaluate the effect of insulin like growth factor-I (rhIGF-I) in complex with binding protein 3 (IGFBP 3) compared to the effect of free IGF-I on muscle protein biosynthesis in undernourished animals. METHODS: Three groups of female Sprague-Dawley rats (200 g) were initially semi-starved for 3 days and then treated with saline (controls), rhIGF I (1 microg g-1) or equimolar amounts of rhIGF-I/rhIGFBP-3 complex (5 microg g-1) i.v. twice daily for 3 days during continuous semistarvation. Protein metabolism in hind limb skeletal muscle was studied by incorporation of L-[14C U]phenylalanine into proteins, western blot determination of translation initiation factors involved in the binding of the 40S ribosomal subunit to mRNA, and quantification of mRNA content for IGF-I, IGF-IR and GH-R. Plasma measurements of insulin, IGF-I and amino acids were also performed. RESULTS: rhIGF-I/rhIGFBP-3, but not rhIGF-I alone, stimulated protein synthesis by 177 +/- 26% (P 0: presence of VRF; MHIS = 0: absence of VRF). As early as 12 weeks, the mean change from the baseline ADAS-Cog score was significantly different for those patients treated with high-dose rivastigmine compared with placebo controls in both MHIS categories. However, the treatment difference between high-dose rivastigmine and placebo at each time-point was larger for patients with MHIS > 0. The proportion of responders was significantly greater in the high-dose rivastigmine group for each level of improvement. No differences were noted between treatment groups regarding safety evaluations. Rivastigmine is effective in both categories of patients, and those with VRF experience greater clinical benefit (cognition, activities of daily living, and disease severity). PMID- 10809937 TI - A population-based study of the associations of stroke occurrence with weather parameters in Siberia, Russia (1982-92). AB - Previous studies have established a seasonal variation in stroke occurrence, but none have assessed the influence of inclement weather conditions on stroke incidence in a general population of Russia. We performed a stroke population based study in the Oktiabrsky District of Novosibirsk, Siberia, Russia. Included in the analysis were 1929 patients with their first occurrence of ischemic stroke (IS), 215 patients with their first occurrence of intracerebral hemorrhage (ICH) and 64 patients with their first occurrence of subarachnoid hemorrhage (SAH): all patients were aged between 25 and 74 years. The cumulative daily occurrence of total strokes and stroke subtypes was evaluated in relation to aggregated daily mean values of ambient temperature, relative humidity and air pressure by means of Poisson regression analysis to estimate the rate ratio (RR) with corresponding confidence interval (CI) and to identify the weather parameters of most importance. In a multivariate analysis, with adjustment for the effects of season, solar and geomagnetic activity, and age of the patients, low ambient temperature (RR 1.32; 95% CI 1.05-1.66) and mean value of air pressure (RR 0.986; 95% CI 0.972-0.999) were important predictors of IS occurrence, while mild ambient temperature (RR 1.52; 95% CI 1. 04-2.22) was an important predictor of ICH occurrence. No relationship between SAH occurrence and any one of the weather parameters studied was revealed. There was no interaction between any meteorological variables that was statistically significant. Inclement weather conditions are associated with the occurrence of IS and ICH in Siberia, Russia. Among the meteorological parameters studied, low ambient temperature and mean air pressure are the most important predictors of IS occurrence, whereas the occurrence of ICH is associated with mild ambient temperature. There is no association between any one of the weather parameters studied and the occurrence of SAH. PMID- 10809938 TI - Are there any unique epidemiological and vascular risk factors for ischaemic strokes that occur in the morning hours? AB - It is now accepted that the incidence of ischaemic stroke is significantly increased in the morning. Any attempts to prevent its occurrence must be based on determining the mechanisms, special risk factors and appropriate protective measures needed during this vulnerable period. We studied the epidemiological features of morning stroke and reviewed the records of 2312 consecutive patients recorded prospectively in the Tel Aviv Stroke Register. Information about time of stroke onset was obtained from the patient, family members or other observers. The study parameters of age, sex, vascular distribution (carotid versus vertebrobasilar), ischaemic heart disease, myocardial infarction, diabetes mellitus, arterial hypertension, smoking, hyperlipidaemia, stroke severity and recurrence were compared between patients with morning stroke and those with stroke occurring at other times. In 599 patients (34%) stroke occurred between 06:00 and 10:00 h. No evaluated parameter was found to be statistically different among the morning stroke patients compared with stroke occurring at other times (P < 0.2). Patients with arterial hypertension and ischaemic heart disease and male patients had a greater likelihood of stroke occurrence between 22:00 and 02:00 h (P < 0.05). Our data suggest that none of the common vascular risk factors could explain the morning peak of stroke occurrence. The next step in the quest for understanding the phenomenon of circadian variation is to identify other physiological factors and the effects of pharmacological agents in morning stroke protection. PMID- 10809939 TI - Prediction of angiographic carotid artery stenosis indexes by colour Doppler assisted duplex imaging. A critical appraisal of the parameters used. AB - The aim of our study was to establish colour Doppler-assisted duplex imaging (CDDI)-criteria to predict an angiographic internal carotid artery (ICA) stenosis of at least 70%, according to the North American Symptomatic Carotid Endarterectomy Trial (NASCET) and European Carotid Surgery Trialists (ECST) methods of measurement. In the following, we describe the findings in 79 patients who were screened for carotid endarterectomy by CDDI and further evaluated by digital subtraction angiography (DSA). In 158 carotid arteries, 107 stenoses of > 30% and nine occlusions were found. Receiver operator characteristic graphs were constructed in order to calculate sensitivities and specificities of the assessment by CDDI in the prediction of high-grade stenoses determined by angiography. Optimal cut-off points were defined by highest accuracy which reflects the combination of high sensitivity and specificity. The critical index of a high-grade ICA stenosis according to the ECST method could be predicted with an accuracy greater than 90% by a systolic peak velocitiy of 1.25 m/s or an area reduction by CDDI of 70%. Corresponding values, 1.6 m/s and 80% area reduction, predicted the stenosis indexes according to the NASCET method less reliably, with accuracies of between 80% and 90%. Flow velocity criterion was slightly less accurate than the area reduction criterion by CDDI. Finally, double-blind evaluation performed by two readers per examination modality showed that the measurement of area reduction in CDDI is at least as reliable as stenosis indexes according to ECST and NASCET methods. PMID- 10809940 TI - Progressive response to botulinum A toxin in cerebral palsy. AB - Botulinum A toxin (BT) has been successfully used for the management of spasticity in cerebral palsy (CP). However, the long-term results of BT have not yet been determined. We have studied the evolution of a homogeneous group of patients with CP treated with BT. All these patients had an equinus gait resulting from calf muscle spasticity without other muscle group involvement. All of these patients were treated with the same total dose (4 microg/kg) at the same time interval (three months). The mean follow-up time was 33 months. Gait evaluation was made blind on videotape recordings by two independent physicians according to five point scale. All our patients exhibited a progressive improvement in their gait pattern. None of our patients developed fixed contractures nor did any of them need surgical correction. No significant side effects were seen. The response observed in our study could be due to a progressive symptomatic effect of BT, but it might be also explained by a change in the natural history of the spasticity related to CP, at least in this selected group of patients. PMID- 10809941 TI - Decreased mRNA expression of TNF-alpha and IL-10 in non-stimulated peripheral blood mononuclear cells in myasthenia gravis. AB - Myasthenia gravis (MG) is characterized by T cell-dependent autoantibodies to the acetylcholine receptor on the post-synaptic membrane of the neuromuscular junction. The production of autoantibodies is regulated by T cells via cytokines. To investigate the involvement of interferon gamma (IFN-gamma), tumour necrosis factor alpha (TNF-alpha), interleukin-4 (IL-4) and interleukin-10 (IL-10) in MG, the mRNA levels of these cytokines in peripheral blood mononuclear cells without stimulation in vitro were quantified by competitive reverse transcription polymerase chain reaction technique. The mRNA level of TNF-alpha was significantly lower (P = 0.0004) in the overall group of MG patients compared with controls. IL-10 was also lower in MG patients (P = 0.008), most markedly in non-thymectomized patients (P = 0.016). There were no significant differences in IFN-gamma and IL-4 between patients and healthy controls, but the mRNA levels of IL-4 in non-thymectomized patients was significantly lower than in controls (P = 0.02) and in thymectomized patients (P = 0.03). These results, reflecting the in vivo expression pattern of these cytokines in the periphery, suggest an altered cytokine expression at the systemic level in MG. PMID- 10809942 TI - Effects of sex hormones on experimental autoimmune myasthenia gravis. AB - To examine whether exacerbation of myasthenia gravis (MG) can be induced by changes in sex hormone levels we immunized 20 female Lewis rats with torpedo antigen to induce experimental autoimmune MG (EAMG). Ten of the animals underwent surgical ovariectomy prior to the induction of EAMG and 10 served as controls. Anti-acetylcholine receptor antibody (AChR-ab) titres and the degree of decrement on repetitive stimulation electromyography (REMG) at 3 Hz were obtained at base line and compared between rats with and without ovariectomy and a second control group of naive rats. Three rats in each group were then injected with excess oestrogen and progesterone for one week, and three of the remaining rats in each group were given sham injections, and the degree of decrement on REMG and AchR-ab titres were re-evaluated. Immune reactivity of peripheral lymphocytes and splenic lymphocytes from all groups and controls was also determined. A comparable number of animals with and without ovariectomy developed clinical and electromyographic EAMG. The extent of decrement on REMG and AChR-ab titres did not change following hormonal replacement. Lymphocyte reactivity was similar for rats with and without ovariectomy. In conclusion, sex hormones do not appear to have an influence on the susceptibility to and the severity of MG. PMID- 10809943 TI - Coexistence of dominant and recessive familial amyotrophic lateral sclerosis with the D90A Cu,Zn superoxide dismutase mutation within the same country. AB - The Cu,Zn superoxide dismutase (Cu,Zn SOD) mutations described in amyotrophic lateral sclerosis (ALS) have, for the most part, a dominant influence. However, while a few cases with a heterozygous D90A mutation have been described in different countries, D90A has been recently proven to be recessively inherited with a common founder effect in Scandinavia. We screened French ALS families for Cu,Zn SOD mutations. The presence of the D90A allele was found in two index cases, and their families were subsequently studied. In the first family the ALS patients were homozygotes for D90A, while in the second, all ALS patients were heterozygotes. In both families the disease was found to initially involve the lower limbs with slower progression than in sporadic cases, and frequent atypical signs such as paresthesia and urgency of micturition. We determined the D90A allele frequency in controls (n = 200) and sporadic ALS patients (n = 408). No D90A allele was found. This is the first report of coexistence of dominant and recessive families with the D90A Cu,Zn SOD mutation within the same country. PMID- 10809944 TI - Alcohol with xylocaine for treatment of eyelid dystonia. AB - Eyelid dystonia represents a form of idiopathic torsion dystonia, and it is considered as a rare disorder. Although botulinum toxin injections into the eyelids and eyebrows are considered by many neurologists as the treatment of choice for this, its cost limit its use in our community. A study was conducted at the King Hussein Medical Center between January 1995 and January 1998 using 1 2 ml of 0.5% xylocaine added to 99.5% ethanol in one-tenth of the volume of xylocaine, made injectable into the eyelids and eyebrows muscles in patients with blepharospasm. The treatment was aimed at reducing muscle spindle afferent activity. Twenty-one patients were given this modality of treatment in at least 8 12 separate sessions, with two weeks elapsing between any two sessions. Mild to moderate improvement was observed in 47.6% of our patients. The average latency from the time of the injection to the onset of improvement was 30-60 min and the average duration of improvement was 5-7 days. The commonest encountered side effect was bruising at the site of injection that resolved spontaneously in less than 5 days. It is obvious that the usefulness of this treatment is limited by the substantially shorter duration of benefit, thus it cannot be considered as an alternative to Botox treatment. It may be useful to investigate whether a higher alcohol concentration is more effective. PMID- 10809945 TI - Effects of oestrogen replacement therapy on pattern reversal visual evoked potentials. AB - As a result of a regression in the ovarian functions, oestrogen level in circulation during the menopause drops to 1/50 of its value in the normal reproductive cycle. Excitatory oestrogen increases the sensitivity of the central nervous system to catecholamines by changing the opening frequency of voltage related L-type calcium channels and augmenting the effect of glutamate; in addition it inhibits the formation of gamma-amino butyric acid (GABA) by the inhibition of glutamate decarboxylase enzyme. It is argued that oestrogen increases transmission in the optic pathways and that oestrogen is responsible for the shorter latency values and higher amplitudes of visual evoked potentials in women. We recorded the monocular pattern reversal visual evoked potentials (PRVEP) of both eyes of 54 post-menopausal women before treatment and of 30 of them after replacement therapy with Tibolon, and of 24 women receiving placebo treatment. The explicit values of P100 latency of right and left eyes before treatment were 98.8 +/- 3.5 and 99.0 +/- 3.3 ms, respectively. The explicit values of P100 latency of right and left eyes after placebo treatment were 98.6 +/- 3.7 and 98.8 +/- 4.0, respectively. The explicit values of P100 latency of right and left eyes after replacement treatment were 94.6 +/- 3.7 and 94.8 +/- 4.0, respectively. We found a statistically significant decrease in the mean PRVEP latencies and a statistically significant increase in mean amplitudes after replacement treatment (P < 0.001) compared with those before treatment and those after placebo treatment. We attributed the changes in PRVEP values after replacement treatment to the action of Tibolon, which acted as a natural sex steroid and speeded the visual transmission time via the widespread receptors in the central nervous system. It is concluded that PRVEP is an objective electrophysiological assessment method in evaluating the efficiency of hormone replacement therapy in post-menopausal women. PMID- 10809946 TI - Acute axonal form of Guillain-Barre syndrome in a multiple sclerosis patient: chance association or linked disorders? AB - Multiple sclerosis (MS) is characterized by inflammation, demyelination and gliosis, involving the central nervous system (CNS) and commonly sparing the peripheral nervous system (PNS). Coexistence of CNS and PNS chronic demyelination has been rarely demonstrated in chronic inflammatory demyelinating polyradiculoneuropathies (CIDP) and in MS, but the occurrence of acute polyradiculoneuropathy in a patient with MS is even more unusual. We describe the case of a woman with relapsing-remitting MS who presented with an acute severe tetraparesis. Cerebrospinal fluid (CSF) examination together with neurophysiological data and sural nerve biopsy study demonstrated an axonal form of Guillain-Barre Syndrome (GBS). It remains unresolved if the association of an axonal form of GBS and MS is fortuitous or, on the contrary, is indicative of the coexistence in some individuals of common pathogenetic mechanisms. PMID- 10809947 TI - Ataxic form of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). AB - We reported a 64-year-old male with an eight-month history of gait disturbance and sensory impairment. The patient initially noticed unsteadiness of gait and numbness in his feet, and these symptoms progressed until he was unable to walk without assistance five months later. Vibratory sensation and position sense were markedly diminished, and deep tendon reflexes were absent in all extremities. Motor conduction velocities were slow with prolonged distal latencies, and sensory nerve action potentials (SNAP) were not elicited. Sural nerve biopsy revealed a mild loss of myelinated fibres and segmental demyelination. Cerebrospinal fluid showed normal cell count with protein 526 mg/dL. Anti-GM1, anti-GM2 and anti-GA1 antibodies in serum were positive. We diagnosed chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) presenting ataxia. Steroid therapy provided immediate improvement of symptoms and signs. This case suggests that CIDP should be considered as one of the potential causes of ataxic neuropathy. PMID- 10809948 TI - Syringomyelia complicating syphilitic spinal meningitis: a case report. AB - An association between syringomyelia and spinal syphilis was described in the early literature but has not been the subject of reports subsequently. We give details of a contemporary case, affirmed by magnetic resonance imaging. The patient showed significant clinical recovery following penicillin treatment, while the imaging appearances became less pronounced. PMID- 10809949 TI - Paroxysmal dystonia induced by exercise and acetazolamide. AB - We report a case of a 40-year-old woman with dystonic attacks precipitated by slight exercise. Episodes lasted 2-5 min and were not precipitated by sudden movements or by being startled, drinking alcohol, coffee or tea, or by stress. Secondary dystonia was ruled out and brain magnetic resonance imaging (MRI) was unremarkable. Routine and video electroencephalogram (EEG) during and between attacks were normal. Acetazolamide greatly worsened her condition, whereas gabapentin [1-(aminomethyl) cyclohexaneacetic acid] treatment markedly reduced the frequency and severity of the episodes. PMID- 10809950 TI - Frosted branch angitis associated with aseptic meningitis. PMID- 10809951 TI - Progressive muscle pain, weight gain, fatique and unusual body shape change. AB - Multiple symmetrical lipomatosis (MSL) is a rare syndrome characterized by symmetric unencapsulated lipomas around the axial region and frequently associated with neurological involvement, particularly myopathy and neuropathy (Klopstock et al., 1994). Here we present a typical image of a patient with MSL, explaining this disease for neurologists who might care for these patients in an average day at the neuromuscular consultation unit. PMID- 10809952 TI - Dendritic cell-B-cell interaction: dendritic cells provide B cells with CD40 independent proliferation signals and CD40-dependent survival signals. AB - Dendritic cells (DC) have recently been shown to play an important role in B-cell function. We have previously shown that DC can capture and retain unprocessed antigen in vitro and in vivo, and can transfer this antigen to naive B cells to initiate antigen-specific antibody responses. We also demonstrated that DC were providing B cells with isotype-switch signals independent of T cells but that T cell help was essential for antibody production. In this study, using B cells and DC from wild type (WT) and CD40 knockout (CD40KO) mice we show that DC initiate proliferation of B cells independently of CD40, because WT or CD40KO DC could induce proliferation of WT or CD40KO B cells, but proliferation was greater in the absence of CD40. DC also provide B cells with survival signals as WT DC improved viability of B cells after a 5-day culture but survival was reduced in the absence of CD40 expression. PMID- 10809953 TI - The role of complement in the acquired immune response. AB - Studies over the past three decades have clearly established a central role for complement in the promotion of a humoral immune response. The primary function of complement, in this regard, is to opsonize antigen or immune complexes for uptake by complement receptor type 2 (CR2, CD21) expressed on B cells, follicular dendritic cells (FDC) and some T cells. A variety of mechanisms appear to be involved in complement-mediated promotion of the humoral response. These include: enhancement of antigen (Ag) uptake and processing by both Ag-specific and non specific B cells for presentation to specific T cells; the activation of a CD21/CD19 complex-mediated signalling pathway in B cells, which provides a stimulus synergistic to that induced by antigen interaction with the B-cell receptor (BCR); and promotion of the interaction between B cells and FDC, where C3d-bearing immune complexes participate in intercellular bridging. Finally, current studies suggest that CR2 may also play a role in the determination of B cell tolerance towards self-antigens and thereby hold the key to the previously observed correlation between deficiencies of the early complement components and autoimmune disease. PMID- 10809954 TI - Lysosomal cathepsin B plays an important role in antigen processing, while cathepsin D is involved in degradation of the invariant chain inovalbumin immunized mice. AB - We previously reported that CA074, a specific inhibitor of cathepsin B, modulates specific immune responses from the T helper 2 (Th2) type to Th1 type in BALB/c mice infected with Leishmania major. In the present study, we found that a similar type of immune deviation was also induced in mice immunized with ovalbumin (OVA). However, treatment of mice with pepstatin A, a specific cathepsin D inhibitor, suppressed the OVA-specific proliferation of lymphocytes and blocked the development of both Th1 and Th2 cellular responses. These inhibitors did not appear to have any direct influence in vitro on functions of naive lymphocytes. OVA antigen (47 000 MW) was digested mainly into 40 000 MW protein in vitro by lysosomal proteases from naive BALB/c mice, and its digestion was markedly inhibited by the addition of CA074, but not by addition of pepstatin A, during incubation. However, pepstatin A strongly suppressed the degradation of the major histocompatibility complex class II-associated invariant chain (Ii) molecule in vivo and in vitro. Thus, cathepsin B appears to process antigens directed to preferential activation of Th2 cells, while cathepsin D may be responsible for the degradation of Ii, the processing of which is essential in initiating the antigen-specific activation of Th1 and Th2 CD4+ T cells. These lysosomal proteases may have different functions in regulating immune responses. PMID- 10809955 TI - Regulation of epidermal Langerhans cell migration by lactoferrin. AB - Lactoferrin (LF) is a member of the transferrin family of iron-binding glycoproteins to which several anti-inflammatory functions have been ascribed. LF has been shown to down-regulate expression of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha), although the possibility has been raised that the activity of LF in this regard was indirect and secondary to its ability to bind to and inactivate the bacterial lipopolysaccharide (LPS) used to induce cytokine production. However, the identification of putative membrane receptors for LF raises the possibility that the interaction of LF with its receptor may be one important route through which this protein exerts anti inflammatory activity. In the present investigations the biological properties of LF have been examined in a model of cutaneous immune function where the allergen induced migration of epidermal Langerhans cells (LC) from the skin and their subsequent accumulation as dendritic cells (DC) in skin-draining lymph nodes are known to be dependent upon the de novo synthesis of TNF-alpha, but independent of exogenous LPS. Consistent with the protein having direct anti-inflammatory properties, it was found that the intradermal injection of recombinant murine LF (either iron-saturated or iron-depleted LF) inhibited significantly allergen (oxazolone) -induced LC migration and DC accumulation. That these inhibitory effects were secondary to the inhibition of local TNF-alpha synthesis was suggested by the findings that first, LF was unable to inhibit LC migration induced by intradermal injection of TNF-alpha itself, and second, that migration stimulated by local administration of another epidermal cytokine, interleukin 1beta, which is also dependent upon TNF-alpha production, was impaired significantly by prior treatment with LF. Finally, immunohistochemical analyses demonstrated the presence of LF in skin, associated primarily with keratinocytes. Collectively these data support the possession by LF of direct immunomodulatory and/or anti-inflammatory activity, probably associated in this case with inhibition of cytokine production. Furthermore, the results suggest that as a constituent of normal skin, LF may play a role in homeostatic regulation of cutaneous immune function. PMID- 10809956 TI - Individual and combined effect of granulocyte-macrophage colony-stimulating factor and prolactin on maturation of dendritic cells from blood monocytes under serum-free conditions. AB - Prolactin (PRL) shares structural and functional features with haemopoietic factors and cytokine peptides. Dendritic cells (DC) are involved in both initiating the primary and boosting the secondary host immune response and can be differentiated in vitro from precursors under the effect of granulocyte macrophage colony-stimulating factor (GM-CSF) plus other factors. Because PRL has been shown to functionally interact with GM-CSF, we have addressed its role on GM CSF-driven differentiation of DC. Monocytic DC precursors from peripheral blood mononuclear cells (PBMC) were enriched either by adhesion to a plastic surface or CD14-positive selection and cultured for 7 days in serum-free medium containing GM-CSF, interleukin (IL)-4 and PRL, alone or in combination. Cells with large, veiled cytoplasm, expressing major histocompatibility complex (MHC) class II and the costimulatory molecules CD80, CD86 and CD40 and lacking the monocyte marker CD14, were considered as having the phenotype of cytokine-generated DC. Functional maturation was assessed by proliferation and interferon-gamma (IFN gamma) release of allogeneic T lymphocytes. Physiological (10-20 ng/ml) concentrations of PRL interacted synergistically with GM-CSF and the effect was similar to that induced by IL-4 on GM-CSF-driven DC maturation. When used alone, the physiological concentrations of PRL were inhibitory, whereas higher concentrations (80 ng/ml) were stimulatory. The synergistic effect of PRL may in part be caused by its ability to counteract the down-modulation of the GM-CSF receptor observed in serum-free conditions. These data provide further evidence of the significance of PRL in the process of T lymphocyte activation. PMID- 10809957 TI - CD1d structure and regulation on human thymocytes, peripheral blood T cells, B cells and monocytes. AB - Human T cells expressing CD161 and an invariant T-cell receptor (TCR) alpha-chain (Valpha24invt T cells) specifically recognize CD1d and appear to have immunoregulatory functions. However, the physiological target cells for this T cell population, and whether alterations in CD1d expression contribute to the regulation of Valpha24invt T-cell responses, remain to be determined. A series of antibodies were generated to assess CD1d expression, structure and regulation on human lymphoid and myeloid cells. CD1d was expressed at high levels by human cortical thymocytes and immunoprecipitation analyses showed it to be a 48 000-MW glycosylated protein. However, after solubilization, the majority of the thymocyte CD1d protein, but not CD1d expressed by transfected cells, lost reactivity with monoclonal antibodies (mAbs) against native CD1d, indicating that it was alternatively processed. Moreover, thymocytes were not recognized by CD1d reactive Valpha24invt T-cell clones. Medullary thymocytes and resting peripheral blood T cells were CD1d-, but low-level CD1d expression was induced on activated T cells. CD1d was expressed by B cells in peripheral blood and lymph node mantle zones, but germinal centres were CD1d-. Resting monocytes were CD1d+ but, in contrast to CD1a, b and c, their surface expression of CD1d was not up-regulated by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) activation. These results demonstrate constitutive CD1d expression by human professional antigen-presenting cells and that post-translational processing of CD1d may contribute to regulation of the activity of CD1d-specific T cells. PMID- 10809958 TI - Exploring the feasibility of an anti-idiotypic cocaine vaccine: analysis of the specificity of anticocaine antibodies (Ab1) capable of inducing Ab2beta anti idiotypic antibodies. AB - Conventional vaccination with the cocaine molecule conjugated to a protein carrier is a new approach in the treatment of addiction. Experimentally, this strategy has been shown to alter the pharmacokinetics as well as the psychostimulant effect of a cocaine challenge. The purpose of this study was to investigate whether a more stable and less controversial molecule, an anti idiotypic antibody, which mimics the configuration of the cocaine molecule (Ab2beta), could be successfully used instead of cocaine. Two cocaine conjugates that presented different areas of the cocaine molecule to the immune system were used to produce monoclonal antibodies specific for cocaine (Ab1). Several anti idiotypic antibodies were then produced. Four were identified as Ab2beta, or internal images of the antigen; when injected into BALB/c mice, they elicited an anticocaine response. The anticocaine response elicited by one of the four Ab2beta (K1-4c) was sufficient to significantly reduce the level of cocaine that targeted the brain following cocaine challenge, compared with the level of cocaine found in the brain of control animals immunized with irrelevant antibody. In conclusion, the possibility of an anti-idiotypic vaccine seems to be worth pursuing. PMID- 10809959 TI - Morphine-induced macrophage apoptosis: the role of transforming growth factor beta. AB - Laboratory and clinical reports indicate that opiate addicts are prone to infections. This effect of opiates is partly attributed to opiate-induced macrophage (Mphi) apoptosis. In the present study, we evaluated the role of transforming growth factor-beta (TGF-beta) in morphine-induced apoptosis of murine J774 cells and peritoneal Mphi. Mphi harvested from morphine-treated mice showed greater (P < 0. 0001) apoptosis when compared with control Mphi. Morphine also enhanced apoptosis of J774 cells and peritoneal Mphi. Anti-TGF-beta antibody inhibited (P < 0.001) the morphine-induced apoptosis in J774 cells (control 0.7 +/- 0.4%; 10-6 M morphine 23.5 +/- 0.7%; anti-TGF-beta antibody (Ab) + 10-6 M morphine 8.1 +/- 0.7%; apoptotic cells/field) and peritoneal Mphi (control 1.5 +/ 0.9%; 10-6 M morphine 29.1 +/- 1.4%; 10-6 M morphine + anti-TGF-beta Ab 19. 1 +/ 1.8%; apoptotic cells/field). TGF-beta enhanced (P < 0.001) apoptosis of J774 cells and peritoneal Mphi. TGF-beta also promoted Mphi DNA fragmentation into integer multiples of 180 bp (ladder pattern). Immunocytochemical studies revealed that morphine enhanced the Mphi cytoplasmic content of TGF-beta. In addition, Western blotting showed increased production of TGF-beta by morphine-treated J774 cells when compared with control cells. Morphine increased J774 cell expression of bax. Interestingly, morphine-induced bax expression was inhibited by anti-TGF beta Ab. As both morphine-induced J774 cell apoptosis and bax expression were inhibited by anti-TGF-beta Ab, it appears that morphine-induced J774 cell apoptosis may be mediated through the generation of TGF-beta. PMID- 10809960 TI - Interleukin-16 stimulates the expression and production of pro-inflammatory cytokines by human monocytes. AB - Interleukin-16 (IL-16) acts as a chemoattractant for CD4+ cells, as a modulator of T-cell activation, and plays a key role in asthma. This report describes the cytokine-inducing effects of IL-16 on total peripheral blood mononuclear cells (PBMC) and PBMC subpopulations. While CD4+ T lymphocytes did not secrete cytokines in response to rhIL-16, CD14+ CD4+ monocytes and maturing macrophages secrete IL-1beta, IL-6, IL-15 and tumour necrosis factor-alpha (TNF-alpha) upon rhIL-16 stimulation. The mRNA species for these four cytokines were detected as early as 4 hr post-stimulation, with protein being secreted by 24 hr. Secretion of IL-1beta and IL-6 by total PBMC was dose dependent, with maximal secretion being observed using 50 ng/ml rhIL-16. However, for IL-15 or TNF-alpha maximal secretion by total PBMC occurred with all concentrations between 5 ng/ml to 500 ng/ml rhIL-16. Purified monocytes/macrophages secreted maximal concentrations of all four cytokines in the presence of 500 ng/ml rhIL-16, except for monocytes where maximal secretion of IL-15 was, interestingly, observed with only 50 ng/ml rhIL-16. The use of higher concentrations of rhIL-16 (1000 ng/ml) inhibited secretion of all four cytokines. While these IL-16-induced cytokines are likely to be involved in the immune system's response to antigen, the data suggest that IL-16 may play a key role in initiating and/or sustaining an inflammatory response. PMID- 10809961 TI - Expression of cytokine genes in Marek's disease virus-infected chickens and chicken embryo fibroblast cultures. AB - The role of cytokines in the pathogenesis and immunity of Marek's disease (MD), a herpesvirus-induced T-cell lymphoma in chickens, is poorly understood. Two different experiments were used to examine the potential role of particular cytokines in the pathogenesis and immune responses of MD. First, chicken embryo fibroblasts (CEF) were stimulated with lipopolysaccharide (LPS) and/or recombinant chicken interferon-gamma (rChIFN-gamma) and used to develop techniques for examining transcription of IFN-alpha, IFN-gamma, inducible nitric oxide synthase (iNOS), interleukin (IL)-1beta, IL-2, IL-6 and IL-8 by reverse transcription-polymerase chain reaction (RT-PCR). Addition of LPS and/or rChIFN gamma resulted in the up-regulation of mRNA for iNOS, IL-1beta and IL-6, while IFN-gamma was up-regulated by LPS alone. IL-2 was down-regulated by the treatments. Second, to determine the effects of Marek's disease herpesvirus (MDV) infection on cytokine transcription in vivo, chickens were infected with MDV at 21 days of age and examined at 7 days post-infection (p.i.) (exp. 1) or were infected with MDV at 1 day of age and examined from 3 to 15 days p.i. (exp. 2). In MDV-infected chickens, IFN-gamma transcription was up-regulated as early as 3 days p.i. until the termination of the experiment at 15 days p.i., while iNOS and IL-1beta were up-regulated between 6 and 15 days p.i. Infection of 1-day-old chicks increased levels of mRNA for IFN-gamma and iNOS between 16- and 64-fold at 9 days p.i. These results suggest that IFN-gamma and iNOS may play an important role in the pathogenesis of MD. PMID- 10809962 TI - Expression and functional activity of the very late activation antigen-4 molecule on human natural killer cells in different states of activation. AB - In the present study we describe the expression and functional activity of the alpha4beta1 heterodimer molecule on human natural killer (NK) cells. Flow cytometric analyses showed that fresh and activated NK cells expressed high levels of very late activation antigen-4 (VLA-4) molecules. These cells bound to fibronectin (FN) and to its 38 000-MW proteolytic fragment through the VLA-4 integrin that was blocked with HP2/1 anti-alpha4 monoclonal antibodies (mAbs) and with the FN peptide fragment CS1. No inhibitory effects were observed in the presence of anti-alpha5 mAb, FN peptide fragment CS2 or other irrelevant mAb. Fresh NK cells were unable to aggregate, despite their expression of VLA-4, and only activated (cultured and lymphocyte-activated killer cells) NK cells showed homotypic aggregation with HP1/7 and HP2/4 anti-alpha4 mAb related to cellular activation. These results underline new evidence of how NK cells in different states of activation maintain different constitutive levels of alpha4beta1 integrin activity, and highlight the possibility of a different functional regulation by the cells bearing VLA-4, in the expression of these epitopes and their ability to interact with their ligands. PMID- 10809963 TI - Deficiency of the macrophage migration inhibitory factor gene has no significant effect on endotoxaemia. AB - By targeted disruption of the MIF gene, we have established a mouse strain deficient in macrophage (Mphi) migration inhibitory factor (MIF). Despite previous reports indicating an essential role of MIF in endotoxaemia, an injection of lipopolysaccharide (LPS) into the MIF-deficient mice (maintained under specific pathogen-free conditions) caused shock. No significant difference was detected between the MIF-deficient mutant and normal mice in susceptibility to LPS for endotoxaemia or tumour necrosis factor-alpha (TNF-alpha) formation upon LPS injection. Peritoneal Mphi from the two strains produced TNF-alpha in response to LPS with similar dose responses. Dexamethasone suppressed the LPS induced TNF-alpha response of Mphi, but no difference was detected between the Mphi from the two strains. These results suggest that endogenous MIF has no significant effect on the LPS-induced TNF-alpha production and no effect on suppression of the response by glucocorticoids. Thus, MIF is not crucial for LPS induced immune responses leading to shock. PMID- 10809964 TI - Decay accelerating factor in guinea-pig reproductive organs. AB - Decay accelerating factor (DAF, CD55) expressed in human reproductive organs and gametes is thought to play a pivotal role in protection against autologous complement activation in the genital tract. To further investigate the role of DAF in reproduction, we analysed DAF distribution in reproductive organs using guinea-pigs that express multiple DAF isoforms. In males, significant staining was observed in testis on the elongated spermatids and spermatozoa. Levels of DAF mRNA with a shorter 3' untranslated region were significantly enhanced in testis from 9 weeks of age, indicating the presence of DAF mRNA and protein synthesis of spermatozoa DAF in late haploid germ cells. Epididymal spermatozoa appeared to express DAF on the inner acrosomal membrane as well as over their entire surface. Significant DAF expression was also observed on the epithelium of seminal vesicles from 4 weeks of age, with no increase thereafter in the mRNA. C3 mRNA was not detected in this tissue. In females, DAF was detected on the plasma membranes of oocytes through follicle development and on the apical region of uterine epithelium, although the levels of DAF mRNA in these tissues were low. In addition, DAF was selectively expressed on the apical region of ciliated oviductal epithelial cells. The apical region of the ciliated cells comprising the efferent ductule epithelium was also stained significantly, even at 12 days of age, while other epididymal epithelial cells were hardly stained at any age, suggesting that DAF is constitutively expressed on cilia. PMID- 10809965 TI - A novel function of B lymphocytes from normal mice to suppress autoimmunity in (NZB x NZW)F1 mice. AB - In systemic autoimmune-prone (NZB x NZW)F1 (NZB/W F1) mice, B-cell abnormalities characterized by hypergammaglobulinaemia accompanying autoantibodies have been thought to be a main cause of the disease. To examine a possible regulatory role of B cells in the disease manifestations, we injected, intravenously (i.v.), normal or autoimmune B cells into non-irradiated NZB/W F1 mice. The injection of splenic B cells from major histocompatibility (MHC)-matched or allogeneic normal mice caused a marked decrease in serum immunoglobulin G (IgG) levels of autoantibodies, delayed the appearance of proteinuria and prolonged life span, whereas treatment with splenic B cells from NZB/W F1 or X-linked immunodeficient (Xid) mice failed to suppress the autoimmunity. Moreover, in vitro polyclonal antibody responses to lipopolysaccharide (LPS) of NZB/W F1-derived B cells from the treated mice were markedly reduced. Interestingly, the treatment of NZB/W F1 mice at 16, 18 and 20 or at 20, 22 and 24 weeks of age was more effective than that at 6, 8 and 10 weeks. The treatment also inhibited the development of surface IgG+ (sIgG+) B cells and splenomegaly, prominent in aged NZB/W F1 mice. In addition, when untreated NZB/W F1 responding B cells were precultured with normal B cells in vitro for 3 days, they also diminished the autoantibody production to subsequent LPS stimulation. Hence, the present results imply a novel function of normal B cells to ameliorate autoimmune disease in NZB/W F1 mice by correcting their B-cell abnormalities, and indicate that NZB/W F1 and Xid mice possess defects in this regulatory B-cell function. PMID- 10809966 TI - Tamoxifen alleviates disease severity and decreases double negative T cells in autoimmune MRL-lpr/lpr mice. AB - Previous study suggested that MRL-lpr/lpr mice treated with tamoxifen (TAM) had less severe proteinuria, reduced serum titre of anti-dsDNA autoantibodies and an increased survival rate. To investigate further the regulatory mechanisms of TAM on MRL-lpr/lpr female mice, a total dose of 200 microg per mice (5.5 mg/kg) was given every 2 weeks subcutaneously, while the control mice were injected with oil only. After being treated with TAM four times, the mice were killed and cellular functions were evaluated. The TAM-treated groups had smaller sized spleen and lymph nodes. Flow cytometric analysis of splenocytes had a significantly lower percentage of cell number of T cells and double negative T cells (CD4- CD8- T cells). There was no difference in cytokine production (interleukin (IL)-2, IL-4, IL-5, IL-10 and interferon-gamma (IFN-gamma)) from splenocytes stimulated with concanavalin A (Con A) or cytokines (IL-6) secreted by peritoneal exudate cells when stimulated with lipopolysaccharide (LPS). However, IL-2 from lymph node cells was significantly higher on TAM-treated mice. Finally, splenocytes or purified T cells stimulated with anti-CD3 antibody plus cross-linking immunoglobulin G (IgG) of the TAM-treated group had higher 3H-incorporation of proliferation assay compared with that of control groups. In vitro study further demonstrated that IL-2-activated proliferation of lymph node double negative (DN) T cells can be inhibited by TAM treatment in a dose-dependent manner. Our finding demonstrated that TAM may potentially influence T cells and modulate the immune function, which offers a novel approach to explore the feasibility of hormone therapy for autoimmune diseases. PMID- 10809967 TI - Antibody repertoire development in fetal and newborn piglets, III. Colonization of the gastrointestinal tract selectively diversifies the preimmune repertoire in mucosal lymphoid tissues. AB - Changes in the VH-region repertoire of isolator piglets reared for 6 weeks under germ-free (GF) conditions and those colonized (COL) with a defined exclusion flora on the 1st day of life were compared. Although serum immunoglobulin levels were 20-100-fold higher in COL piglets than GF piglets, an analysis of peripheral blood B cells (PBBs) indicated that: GF and COL piglets used the same four VH genes and two DH segments during the 6-week period; proportional usage of VH genes and DH segments was the same as in fetal animals; and VH and DH usage did not differ between COL and GF animals. This pattern differed from the PBBs from 6 week-old conventional (CONV) piglets. When the sequences of 73 splenic CDR3 segments were analysed, DH usage and mutation frequency were the same in sequences from both 6-week-old GF and COL piglets; mutations were infrequent and occurred with the same frequency as in 110-day fetal spleen. However, the median CDR3 length in COL piglets was shifted upward due to 3' DH N-nucleotide additions. Neither COL nor GF animals made specific serum antibodies to phosphoryl choline given parenterally on a T-cell dependent carrier. In contrast to the near absence of a colonization effect in PBBs and splenic DNA, rearranged variable heavy-chain gene segments (VDJs) recovered from the DNA of mucosal lymphoid tissues of COL piglets showed pronounced differences from those recovered from GF animals in usage of DHA-, DHB-and VHB- and in the frequency of point mutation. The mucosal VDJ transcripts and those from the spleen were similarly affected by colonization. This effect on mucosal lymphoid tissue was consistent with the five-fold selective increase in serum immunoglobulin A (IgA) levels relative to IgM and IgG. Comparison of IgM and IgA transcripts from mucosal tissues suggested that IgA and IgM clones diversify in parallel. Our findings are the first to show that colonization of the gastrointestinal tract of offspring separated from their mothers, differs from 'conventionalized' GF animals in that colonization preferentially influences diversification and expansion of the preimmune IgM and IgA repertoire in mucosal lymphoid tissues but not in PBBs and seldom/modestly in VDJs from splenic DNA. PMID- 10809969 TI - Onychomycosis: clinical perspective. PMID- 10809970 TI - The effects of drugs on wound healing: part 1. PMID- 10809968 TI - Molecular remodelling of human CD46 for xenotransplantation: designing a potent complement regulator without measles virus receptor activity. AB - In pig-to-human discordant xenotransplantation, human complement (C) is a major barrier to long survival of xenografts. The current idea on how to cope with this barrier is that human complement regulatory proteins are forcibly expressed on xenografts to serve as safeguards against host C-induced hyperacute rejection of xenografts. Co-expression of decay-accelerating factor (DAF) (CD55) and membrane cofactor protein (MCP) (CD46) would be the first choice for this trial, because most of the human cells are protected from C-mediated damage by two different modes with these two kinds of C-regulators. Many problems have arisen, however, for MCP expression on grafts. (i) MCP acts as a measles virus receptor, which may function to render donor pigs measles virus (MV) sensitive. (ii) MCP signals immune suppression which causes devastation of the recipient's immune responses. (iii) MCP exerts relatively low self-protective activity against C compared with other cofactors; development of more efficient forms is desirable. (iv) Grafts with a high expression level of MCP are difficult to produce. In this study, we made a number of cDNA constructs of MCP, expressed them on swine endothelial cell lines, and tested cell-protective potency and MV susceptibility. The short consensus repeat 1 (SCR1)-deleted MCP with glycosyl phosphatidylinositol (GPI) anchored form (Delta1MCP-PI) of MCP was found to be most suitable for the purpose of overcoming these problems. However, it was also found that MV induces two modes of cytopathic effect (CPE) on swine endothelial cells, either MCP-dependent or -independent. Here, we discuss these two points which will be raised through study of MCP-transgenic animals. PMID- 10809971 TI - Yaws-a review of the last 50 years. PMID- 10809972 TI - Treatment of tinea capitis. PMID- 10809973 TI - Tinea capitis in Benghazi, Libya. AB - BACKGROUND: Tinea capitis is a worldwide problem. It affects mainly school age children. Late detection and improper treatment of the inflammatory type of this disease may result in disfigurement and permanent alopecia. The objectives of this study were to determine the prevalence, clinical types, and causative species of tinea capitis in Benghazi, Libya. METHODS: One hundred and ninety six patients with tinea capitis were enrolled in this study. Hair stumps and scales were collected from every case and exposed to direct microscopic examination using 10% potassium hydroxide solution, and cultivation on modified Sabouraud's dextrose agar with cyclohexamide and chloramphenicol. RESULTS: Tinea capitis accounts for 45% of all superficial fungal infection and 92% occurred in children below the age of 10 years. The gray patch type was the most common clinical variety (53.6%), followed by black dots, seborrhoid type, and kerion (25.5%, 10.2%, and 8.2%), respectively. Four patients with a clinical picture of alopecia areata-like lesion and one patient with a favus-type lesion were seen. Species identification revealed that Trichophyton violaceum was the most common causative agent, responsible for 49.4% of infection, followed by Microsporum canis (38.6%) and T. verrucosum (7.8%). From seven patients the isolate was a mixture of both T. violaceum and M. canis. CONCLUSIONS: There has been a dramatic decrease in the incidence of favus with complete disappearance of T. schoenleinii. T. verrucosum as a causative agent of tinea capitis in this area has been reported for the first time in this study. PMID- 10809974 TI - Onychomycosis and Trichosporon beigelii in Korea. AB - BACKGROUND: Onychomycosis is a common superficial fungal infection. Causative organisms in onychomycosis have been extensively studied, but the role of nondermatophytes is controversial. Trichosporon beigelii is a soil and water inhabiting yeast and is occasionally found in the flora normally associated with human skin, mouth, and nails. Several reports in the literature have suggested that T. beigelii is one of the pathogens in onychomycosis. METHODS: We performed a survey of the mycologic laboratory records of patients clinically suspected of having onychomycosis from July 1996 to December 1998. RESULTS: Out of a total of 2591 nail samples examined, 1222 (47.2%) were culture positive, including 262 cases (10.1%) with T. beigelii. The overall positive rate for the KOH mount examination was 58.8%, and in the cases with T. beigelii was 89.1%. Among the age groups, the incidence rate was highest in the fifth decade (26.6%). The most common causative organism of microscopy-positive onychomycosis was Trichophyton rubrum (61.4%); the others in decreasing frequency were T. beigelii (20.4%), Candida spp. (7.3%), Trichophyton mentagrophytes (4.1%), and mixed infection (2.9%). T. beigelii was repeatedly isolated in successive nail cultures from 10 of 20 patients selected from those with T. beigelii nail infection. CONCLUSIONS: T. beigelii was the second most commonly isolated fungus in onychomycosis and had a high positive rate on KOH mount examination of the nails and successive repeated cultures. We suggest that T. beigelii might be a common pathogen of onychomycosis in Korea. PMID- 10809975 TI - Pseudomonas aeruginosa folliculitis after shower/bath exposure. AB - BACKGROUND: Pseudomonas aeruginosa folliculitis (PF) can develop after exposure to contaminated water in heated swimming pools, whirlpools, and hot-tubes, or after diving suit dressing. METHODS: We observed and studied 14 cases of PF after shower/bath exposure, an underestimated pathogenic event. Cutaneous and environmental microbiological evaluations were performed. RESULTS: In our cases, the clinical expression of dermatitis was constant, PF being a clinically well recognizable skin infection, presenting with follicular, macular, and papulopustular lesions located on the lateral aspect of the trunk, axillary folds, hips, buttocks, and suprapubic area. In all cases, Pseudomonas aeruginosa was isolated from lesional skin; seven cases were serotyped revealing, in three cases, serotype 0 : 1, in two cases 0 : 8, in one case 0 : 10, and in one case 0 : 11. In three families, Pseudomonas aeruginosa was isolated in the well water. In a further three families, Pseudomonas aeruginosa was isolated from bathroom and kitchen components. CONCLUSIONS: Based on our experience, we suggest that shower/bath exposure should be definitively included amongst the possible pathogenic events causing PF. Pseudomonas aeruginosa is responsible for a number of clinical pictures, e.g. otitis externa, conjunctivitis, toe web intertrigo, green nail syndrome, infection of burns and wounds, and folliculitis. Pseudomonas aeruginosa folliculitis (PF) has been reported to develop as a consequence of exposure to contaminated water in heated swimming pools, whirlpools, and hot tubes, or related to diving suits and leg waxing.1-4 We observed 14 cases of PF after shower/bath exposure. This is probably an underestimated pathogenic event; to our knowledge, only one case has been reported to date.5 In our patients, the clinical expression of dermatitis was constant, PF being a clinically well recognizable skin condition. PMID- 10809976 TI - Mast cells in leprosy and leprosy reaction. AB - BACKGROUND: Mast cells can be visualized in routine, acid-fast-staining, paraffin tissue section as metachromatic staining cells, and can be activated to release inflammatory mediators which play a role in the cell-mediated immune response. METHODS: Skin biopsy tissues were taken from the most active skin lesion of each leprosy patient at the time of diagnosis (nonreactional group) and at the time of reaction (reactional group) during the years 1994-1997 in the leprosy clinic at the Department of Dermatology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Thailand. Mast cells were identified by metachromatic staining (purple) in Fite's stain sections and reported as the average number of cells per high power field in three compartments: at the center and periphery of the granuloma and in the interstitium. The data were analyzed in three groups: nonreactional group, type I, and type II leprosy reactions. The mast cell count of each group and each compartment of the section, expressed as the mean +/- standard error, was compared. RESULTS: A total of 95 persons were included in the study, but 108 tissue sections were obtained due to nine cases having more than one section. Of these patients, 63 cases (66.32%) had no reaction, 19 cases (20%) had type I reaction, and 13 cases had type II reaction. There was no difference in age and sex among these groups. The mast cell count in the interstitium was higher than that within the granuloma, both at the center and at the periphery, in every type, and the count in this area reduced significantly in leprosy reactions, both type I and type II, compared with the nonreactional group. CONCLUSIONS: The change in the average mast cell number in nonreactional leprosy and leprosy reactions may indicate the important role of mast cells in dynamic changes in the cell-mediated immune response in leprosy and leprosy reactions. PMID- 10809977 TI - Evidence for Borrelia burgdorferi in morphea and lichen sclerosus. AB - BACKGROUND: Borrelia burgdorferi (Bb) infection has been implicated in the development of morphea and lichen sclerosus; however, conflicting results have been reported with different investigational methods from different regions. We looked for evidence of Bb in patients with morphea and lichen sclerosus by polymerase chain reaction (PCR) analysis of of skin biopsy samples. METHODS: Formalin-fixed, paraffin-embedded skin biopsy samples from 10 patients with morphea and 12 patients with lichen sclerosus were investigated by PCR analysis for the presence of Bb. RESULTS: The presence of Bb DNA was demonstrated in three of 10 patients with morphea and six of 12 patients with lichen sclerosus by nested PCR. CONCLUSIONS: The data obtained in this study suggest that Bb may play a role in the etiopathogenesis of both morphea and lichen sclerosus at least in the western parts of Turkey. PMID- 10809978 TI - A Japanese case of Kindler syndrome. AB - A 25-year-old Japanese woman presented with contracture of the fingers and toes, and difficulty in opening her mouth. Her grandparents are first cousins, but none of the other members of the family are affected. Bulla formation started at birth on areas of the skin that received pressure, and in infancy and early childhood the lesions were limited only to the acral areas. She also had bilateral, incomplete syndactylies involving all web spaces (Fig. 1a). The formation of blisters ceased after the age of 15 years, but a generalized progressive poikiloderma then appeared with accompanying cutaneous atrophy of the skin of the neck, trunk, and extremities (Fig. 1b). The patient experienced mild photosensitivity of the face and neck. At age 18 years, surgical removal of the webbing of all her fingers was performed. Oral examination showed atrophy of the buccal mucosa, and an inability to fully open the mouth. The patient also suffered from poor dentition and easily bleeding gums, but had no symptoms of esophageal dysfunction. Histology of separate biopsy specimens, taken from the poikilodermatous pretibial and trunk skin, showed classical features of poikiloderma, namely epidermal atrophy with flattening of the rete ridges, vacuolization of basal keratinocytes, pigmentary incontinence, and mild dermal perivascularization (Fig. 2a). Interestingly, dyskeratotic cells (Fig. 2b) and eosinophilic rounded bodies (colloid bodies) (Fig. 2c) were frequently found at the basal keratinocyte layer and in the upper dermis, respectively. Pigment was also present in the upper epidermis. To rule out the possibility of a congenital epidermolysis bullosa, ultrastructural and immunofluorescence studies were performed. Ultrastructural studies demonstrated the reduplication of the basal lamina with branching structures within the upper dermis and cleavage between the lamina densa and the cell membrane of the keratinocytes (Fig. 3a). The numbers of associated anchoring fibrils did not seem to be reduced, and colloid bodies and dyskeratotic cells were detected. Immunofluorescence studies with the antibody against type VII collagen (LH 7 : 2) were subsequently carried out. The results showed extensive broad bands with intermittently discontinuous and reticular staining at the dermo-epidermal junction (DEJ) (Fig. 3b), whereas a linear distribution is typically seen in healthy tissue (data not shown). Interestingly, direct immunofluorescence studies revealed intracellular accumulation of immunoglobulin G (IgG), IgM, IgA, and C3 in colloid bodies under the basement membrane (Fig. 3c). PMID- 10809979 TI - A child with spider bite and glomerulonephritis: a diagnostic challenge. AB - A previously healthy 7-year-old white boy presented to St. Louis Children's Hospital with a 1-day history of headache, malaise, temperature of 38.7 degrees C, and a progressively erythematous, tender calf with central dusky purpura. On the morning of admission, his mother noticed a 2-mm crust on the patient's right calf with a 3-cm x 3-cm area of surrounding erythema. No history of recent trauma or bite was obtained. He had suffered two episodes of nonbloody, nonbilious emesis during the last day. In addition, over the previous 12 h, he presented brown urine without dysuria. His mother and brother had suffered from gastroenteritis over the previous week without bloody diarrhea. On initial physical examination, there was a 6-cm x 11-cm macular tender purpuric plaque with a central punctum on the right inner calf, which was warm and tender to the touch, with erythematous streaking towards the popliteal fossa (Fig. 1). The inguinal area was also erythematous with tender lymphadenopathy and induration, but without fluctuance. Laboratory studies included an elevated white blood cell count of 20, 800/microL with 6% bands, 86% segs, and 7% lymphocytes, hemoglobin of 12.5 g/dL, hematocrit of 35.1%, and platelets of 282,000/microL. The prothrombin time/activated partial tissue thromboplastin was 10. 4/28.0 s (normal PT, 9.3-12.3 s; normal PTT, 21.3-33.7 s) and fibrinogen was 558 mg/dL (normal, 192-379 mg/dL). Urinalysis showed 1+ protein, 8-10 white blood cells, too numerous to count red blood cells, and no hemoglobinuria. His electrolytes, blood urea nitrogen (BUN), and creatine were normal. The urine culture was negative. Blood culture after 24 h showed one out of two bottles of coagulase negative Staphylococcus epidermidis. The patient's physical examination was highly suggestive of a brown recluse spider bite with surrounding purpura. Over the next 2 days, the surrounding rim of erythema expanded. The skin within the plaque cleared and peeled at the periphery. The coagulase negative staphylococci in the blood culture were considered to be a contaminant. Cefotaxime and oxacillin were given intravenously. His leg was elevated and cooled with ice packs. The patient's fever resolved within 24 h. The lesion became less erythematous and nontender with decreased warmth and lymphadenopathy. The child was discharged on Duricef for 10 days. Because the patient experienced hematuria rather than hemoglobinuria, nephritis was suggested. In this case, poststreptococcal glomerulonephritis was the most likely cause. His anti-streptolysin-O titer was elevated at 400 U (normal, <200 U) and C3 was 21.4 mg/dL (normal, 83-177 mg/dL). His urine lightened to yellow-brown in color. His blood pressure was normal. Renal ultrasound showed severe left hydronephrosis with cortical atrophy, probably secondary to chronic/congenital ureteropelvic junction obstruction. His right kidney was normal. PMID- 10809980 TI - Cutaneous manifestations of marantic endocarditis. AB - A 70-year-old patient with a history of hypertension and hypercholesterolemia was referred for evaluation of necrotic toes. The patient had a history of several cerebrovascular accidents during the previous month. Initially, she developed sudden-onset left upper extremity weakness which, over the ensuing 4 days, progressed to complete left-sided weakness. This was followed by the development of acute dysarthria. A transesophageal echocardiogram revealed moderate left ventricular hypertrophy, several vegetations on her tri-leaflet aortic valve associated with moderate aortic regurgitation, and a large right atrial thrombus with a mobile component. Bubble studies failed to reveal any septal defects. The patient's electrocardiogram was nonspecific. As serial blood cultures were negative despite fevers of up to 39.8 degrees C, the patient was treated with a 6 week course of intravenous ceftriaxone, ampicillin, gentamicin, and ciprofloxacin for a presumed diagnosis of culture-negative endocarditis. Fungal cultures of the blood were negative. The patient, however, progressed and developed several necrotic toes. Physical examination was significant for ischemic changes of the left first, second, third, and fifth toes, as well as the right first and second toes. Diffuse subungual splinter hemorrhages in the toenails, numerous 2-4-mm palpable purpuric papules on the lower extremities, and nontender hemorrhagic lesions of the soles were also noted. Peripheral and carotid pulses were intact and no carotid bruits were heard. Cardiopulmonary and abdominal examinations were unremarkable. Neurologic examination revealed a disoriented, dysarthric patient with left central facial nerve paralysis, as well as spasticity, hyperactive reflexes, and diminished strength and sensation in the left upper and lower extremities. A left visual field defect and left hemineglect were also present. The patient's last brain computerized tomogram revealed areas of low attenuation consistent with cerebral infarctions in three distinct areas of the brain. These included the left occipitotemporal area, the right parieto-occipital area, and the right posterior frontal region. The regions affected were in the distribution of both the anterior and posterior circulation. No evidence of hemorrhage was noted. The patient subsequently complained of abdominal discomfort. A computerized tomogram of the abdomen with oral and intravenous contrast revealed a 4-cm x 3-cm irregular mass in the tail of the pancreas with several low attenuation lesions throughout the liver which were consistent with infarctions or metastases. Several splenic infarctions were also present. A biopsy of the tumor revealed pancreatic adenocarcinoma. The patient's carcinoembryonic antigen level was 18. 4 ng/mL (0-3) and the CA 19-9 antigen level was 207,000 U/mL (0 36). The alpha-fetoprotein level was normal. Other significant laboratory findings included a prothrombin time of 16.7 (international normalized ratio, 1.4), an activated partial thromboplastin time of 32 (ratio, 1.3), and a platelet count of 85,000/mm3. The Russell viper venom time, sedimentation rate, and C3 levels were normal, and the patient was negative for antinuclear antibodies, anticardiolipin antibodies, and antibodies to extractable nuclear antigens. Of note, the patient was not receiving any anticoagulation. Blood cultures for mycobacteria and fungi, human immunodeficiency virus serology, and urinalysis and culture were negative. The patient subsequently developed an inferior wall myocardial infarction and was transferred to the coronary care unit. In line with the family's request, aggressive care was ceased and the patient expired. The patient's family refused an autopsy. PMID- 10809981 TI - Cutaneous alternariosis in a patient with idiopathic pulmonary fibrosis. AB - A 78-year-old farmer presented with symptomless skin lesions for evaluation. Two years prior, he had developed idiopathic pulmonary fibrosis (IPF) and had been treated thereafter with oral prednisolone 20 mg/day and occasionally with colchicine 1 mg/day. On examination, erythematoviolaceous, slightly infiltrated plaques, measuring approximately 5 x 9 cm, rubbery in consistency, intermingled with pustules, sometimes eroded, with distinctive borders, were noted on the dorsum of both hands and on the extensor surface of both forearms. The lesions had developed over a 20-day period. The skin of these areas was atrophic or eroded with multiple ecchymoses (Fig. 1). The abnormal laboratory findings included an elevated white blood cell count of 17,100/mm3, with 79% neutrophils, 16% lymphocytes, and 5% monocytes, C-reactive protein of 33.15 mg/dL (normal, <0.8 mg/dL), and immunoglobulin G of 598 mg/dL (normal, 701-1545 mg/dL). Other blood and urine tests performed were within normal limits. The diagnosis of IPF was reconfirmed through radiology, high-resolution computed tomography, and spirometry, as well as bronchoscopy and bronchoalveolar lavage fluid analysis. Coexistence of presumptive pulmonary alternariosis was excluded. Hematoxylin and eosin stained sections of the excised cutaneous specimen showed focal ulceration of the epidermis adjacent to a mainly intradermal abscess cavity. Within the latter, remnants of a partly destroyed hair follicle were seen amongst degenerating polymorphonuclear leukocytes, as well as many histiocytes and a few Langhans-type multinucleated giant cells. Minute collections of polymorphonuclear leukocytes were seen in the adjacent epidermis. Periodic acid-Schiff (PAS) and Gomori's silver methenamine stains showed a multitude of broad branching fungal hyphae and large spores within the aforementioned cavity, both free and within the cytoplasm of giant cells (Fig. 2). Immunohistochemistry was performed by means of the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. Sections showed that the infiltrate consisted of an almost equal number of B and T lymphocytes, whereas histiocytes and the few giant cells were labeled with anti CD68 antibodies. Skin smears and biopsy specimens taken twice from all lesions were used for mycologic examination. Wet mounts revealed numerous, brownish, septate hyphae and ovoid Skin smears and biopsy specimens taken twice from all lesions were used for mycologic examination. Wet mounts revealed numerous, brownish, septate hyphae and ovoid structures. Biopsy material was plated on Sabourand's dextrose agar with cloramphenicol (0.05 mg/mL). After 7 days at 27 degrees C, dark, gray-white colonies with a dark brown underside appeared. Microscopic examination of the colonies revealed hyphae with typical conidia having transverse and longitudinal septa. Based on macroscopic and microscopic examination, the isolates were identified as Alternaria alternata (Fig. 3). Treatment with prednisolone was reduced to 10 mg/day and the patient received oral itraconazole (200 mg/day). This resulted in progressive improvement of alternariosis, and the lesions healed completely within 3 months, when treatment was interrupted. Two years later, there is no evidence of recurrence. PMID- 10809982 TI - Cutaneous infestation by Tunga penetrans. PMID- 10809983 TI - Topical liquiritin improves melasma. AB - Twenty women with a clinical diagnosis of melasma were treated with liquiritin cream on one side of the face and with a vehicle cream on the other side twice daily for 4 weeks. Patients were advised to avoid sun exposure and/or used topical sunscreen during the entire period of treatment. Inclusion criteria included an age range from 18 to 40 years and bilateral and symmetrical idiopathic epidermal melasma. Exclusion criteria included patients with dermal melasma (differentiated by Wood's light), melasma with pregnancy, and patients currently receiving hormone replacement therapy. Melasma pigmentary intensity was rated on a five-point scale in relation to the patient's normal facial skin (1, no difference; 2, slightly more pigmented; 3, moderately more pigmented; 4, markedly more pigmented; and 5, intensely more pigmented). The size of the lesions was measured directly using a millimeter grid scale. Ratings and measurements were made prior to treatment and at each of the follow-up visits (after 2, 4, 6, 8, and 10 weeks). Clinical evaluation was performed at week 4; the overall response was rated as excellent, good, fair, or poor. Color photographs were taken at the start and at week 4 of the study. Side-effects were observed and treated. PMID- 10809985 TI - History of dermatology in Guatemala. PMID- 10809984 TI - Successful treatment of tinea capitis with 2% ketoconazole shampoo. AB - BACKGROUND: Oral antifungal drugs are required for effective treatment of tinea capitis. Topical antifungal shampoo's, namely ketoconazole 2% shampoo or products with selenium sulfide or salicylic acid are recommended as adjunctive therapy. Topical antifungal monotherapy has not been successful in the treated of tinea capitis. The purpose of this open study was to evaluate ketoconazole 2% shampoo as a monotherapy for the treatment of tinea capitis. METHOD: A total of 16 black children, aged 3-6, all with proven tinea capitis caused by Trichophyton tonsurans, were treated daily for 8 weeks with 2% ketoconazole shampoo for a total of 56 treatments. Clinical and mycologic examinations were performed every 2 weeks and again at 4 weeks following treatment. The number of colonies were counted on each plate after each visit. Patients with positive cultures after 8 weeks were placed on oral griseofulvin; those with negative cultures were followed monthly by culture for an additional 12 months. RESULTS: Marked clinical improvement occurred in all patients within 2 weeks and absence of pruritus was noted by the patients as early as 2-6 days. After 8 weeks of shampoo, 14 of the 15 (93%) children were clinically healed. Mycologically, the cultures dropped from a confluent growth of T. tonsurans to less than 100 colonies within 2 weeks; fewer than 50 at week 4 and 20 colonies or fewer after week 6. At 8 weeks of treatment the number of colonies remained at 20 or fewer. Six of the 15 children (40%) had negative cultures after 2, 4, and 6 weeks. One child relapsed at the first 4-week follow-up visit. Five of 15 (33%) of the children remained culturally negative for 12 months post-treatment. CONCLUSIONS: Ketoconazole 2% shampoo alone reduces the number of viable arthroconidia in children with tinea capitis thus reducing the transmissibility and contagious nature of the disease. Unexpectedly, complete cure was obtained in 5/15 (33%) of the children. The children remained clinically and mycologically clear as long as one year after treatment. PMID- 10809986 TI - Panniculitis of the breast preceding presentation of Wegener's granulomatosis by 7 years. PMID- 10809987 TI - The utility of submitting epidermoid cysts for histologic examination. PMID- 10809988 TI - Use and misuse of sunscreens. PMID- 10809989 TI - Severe mucosal involvement in a patient with cutaneous leishmaniasis from Nepal. PMID- 10809990 TI - Cockade nevi and spinal dysraphism. PMID- 10809991 TI - Multidrug resistance in haematological malignancies. AB - The development of refractory disease in acute myeloid or lymphoblastic leukaemias (AML, ALL) and multiple myeloma (MM) is frequently associated with the expression of one or several multidrug resistance (MDR) genes. MDR1, MRP1 and LRP have been identified as important adverse prognostic factors in AML, T-ALL and MM. Recently, it has become possible to reverse clinical multidrug resistance by blocking P-glycoprotein-mediated drug efflux. The potential relevance of these reversal agents of MDR and potential new approaches to treat refractory disease are discussed. PMID- 10809992 TI - Hepatitis C virus risk: a hepatitis C virus related syndrome. AB - BACKGROUND: The association between mixed cryoglobulinemia (MC) and hepatitis C virus (HCV) infection has been recently described in many reports. OBJECTIVE: The aim of this study was to evaluate the long-term prognosis of hepatitis C virus positive patients affected by mixed cryoglobulinemia with or without kidney involvement. PATIENTS: At total of 119 hepatitis C virus-positive patients affected by mixed cryoglobulinemia were divided in two groups. Group A: mixed cryoglobulinemia without kidney involvement (103 cases); group B: mixed cryoglobulinemia with glomerulonephritis (GN) (16 cases). A further 37 patients affected by mesangio-proliferative glomerulonephritis (MPGN) were evaluated as controls (group C). METHODS: Anti-hepatitis C virus antibodies were determined by commercial kits and hepatitis C virus-RNA was detected by polymerase chain reaction (PCR) amplification of the 5' untranslated region (5'UTR) of the virus. The hepatitis C virus genotype was determined according to Okamoto. Liver biopsy was performed in 62 patients, bone marrow biopsy in 65 patients, and kidney biopsy in all patients with proteinuria. RESULTS: In group A, 46 patients (45%) were affected by chronic liver disease (CLD), 21 (20%) by low-grade non-Hodgkin's lymphoma (NHL) and 16 (15%) by both diseases. All patients of group B were affected by type I membrano-proliferative glomerulonephritis, 3 (19%) by chronic liver disease, 6 (37%) by low-grade non-Hodgkin's lymphoma, and 7 (44%) by both diseases. Several genotypes of hepatitis C virus were found, but Type 1b was prevalent. In group C, no patient showed chronic liver disease or non-Hodgkin's lymphoma. Younger age, higher mean blood pressure, lower C4 serum level, and poorer survival significantly distinguished group B from group A. Survival rates at 5 years were: 87.4% for group A, 89.5% for group C, and 50.0% for group B. None of the patients of group B developed kidney failure requiring dialysis, whilst infections were the leading cause of death. CONCLUSIONS: In hepatitis C virus-positive patients, the presence of mixed cryoglobulinemia associated with kidney involvement seems to indicate a new syndrome characterized by immune system impairment, lack of progression to kidney failure, and poor survival (hepatitis C virus-Risk syndrome). PMID- 10809993 TI - Myocardial hypertrophy in transgenic mice overexpressing the bovine growth hormone (bGH) gene. AB - OBJECTIVES: The main purpose of the present study was to characterize cardiac muscle hypertrophy using both qualitative and quantitative microscopy in mice overexpressing the bovine growth hormone. RESULTS: Measurements of 30 fibres from each group revealed that fibre diameter in transgenic hearts was significantly larger than in control hearts. There was a significant decrease in interfibrillar space in transgenic hearts as compared with control hearts. The enlarged transgenic hearts displayed unchanged organelles such as normal myofibrils and mitochondria in a normal pattern, suggesting balanced growth. Myelin structures were occasionally observed between normal myofibrils. Moreover, myocardial beta adrenergic receptors and muscarinic receptors in the hearts of transgenic mice overproducing GH were studied to see whether they are involved in the hypertrophic process. It was shown that the density of muscarinic receptors had decreased and the super-high affinity of muscarinic receptors was lost, without any significant changes in either the density or the affinity of beta-adrenergic receptors, as compared with controls. CONCLUSIONS: These results demonstrate that a GH excess was able to induce significant myocardial hypertrophy and that there was a downregulation of muscarinic receptors. PMID- 10809994 TI - Comparison of long versus short duration of anticoagulant therapy after a first episode of venous thromboembolism: a meta-analysis of randomized, controlled trials. AB - OBJECTIVE: To assess the length of oral anticoagulant therapy (short versus long duration) after a first episode of venous thromboembolism (VTE). DESIGN: Meta analysis of randomized controlled trials, comparing two durations of anticoagulation, identified in 1999 by a computerized search of the Cochrane Controlled Trial Register, Medline and Embase, completed by an extensive review of the references of pertinent articles. SETTING AND SUBJECTS: The meta-analysis was performed on literature data. Seven published controlled trials were included. Relative risks with 95% confidence intervals were computed using the relative risk logarithm method. Statistical significance was set up at 0.01 for the test of association. MAIN OUTCOME MEASURES: Outcomes are major haemorrhage and recurrence after a 12-month follow-up. RESULTS: For the recurrence end-point (sample size of 2304 patients), a duration treatment of 12-24 weeks seems preferable to a 3-6 week regimen, with a relative risk (RR) of 0.60 (95% CI: 0.45 0.79, P < 0.001). For the major haemorrhage end-point (1823 patients), the RR is not significantly different from 1 (RR = 1.43, 95% CI: 0.51-4.01, P = 0. 5). The results were similar for the subgroup 'permanent risk factors' or 'idiopathic VTE' (RR for recurrence = 0.48, 95% CI: 0. 34-0.68, P < 0.001). The tendency was similar, although not reaching statistical significance, for the 'temporary risk factors' subgroup (RR for recurrence = 0.34, 95% CI: 0.13-0.93, P = 0.035). CONCLUSIONS: After a first episode of VTE, a long-term treatment regimen allows a significant reduction in the incidence of recurrences without increasing the incidence of bleeding events. PMID- 10809995 TI - Bezafibrate and simvastatin combination therapy for diabetic dyslipidaemia: efficacy and safety. AB - OBJECTIVE: To determine the efficacy and safety of a statin-fibrate combination in diabetes patients. DESIGN.: An open 21-month trial in which each patient first received the single drug for 6 months and then a combination of the two for 1 year. SETTING: Three lipid clinics in university-based tertiary care hospitals. PATIENTS: One hundred and forty-eight patients with type 2 (non-insulin dependent, NIDDM) diabetes mellitus under stable control for 3 months by means of diet and oral hypoglycaemic medication. INTERVENTION: Patients from one clinic (n = 48) received bezafibrate slow release (400 mg day-1), and patients from the other two clinics (n = 100) received simvastatin 20 mg day-1. Six months later, all patients were switched to a daily combination of 400 mg bezafibrate slow release and 20 mg simvastatin for 1 year. RESULTS: The combination of statin and fibrate led to a 23% reduction in total cholesterol, 42% reduction in triglycerides, 29% reduction in LDL-c, 25% increase in HDL-c, 10% decrease in fibrinogen and 19% reduction of Lp(a) levels, and a decrease in the cholesterol/HDL-c ratio (from 8.9 to 5.4) in all 148 patients. Cardiovascular (CV) event rate was significantly reduced from 9.5% during the first 6 months of the study to less than 2% during the last year of the study (whilst on combination Rx). Side-effects with all treatments included only two patients who developed myopathy when on the combined regimen and one on the single statin regimen. However, plasma creatinine phosphokinase (CPK) levels doubled (but remained within the normal range) in most of the patients on combination therapy, compared with only a mild increase in patients receiving a single medication. CONCLUSIONS: The statin and fibrate combination was found to be more efficacious than a single medication for treatment of diabetic dyslipidaemia, as evidenced by improvement in the lipoprotein profile, reductions in Lp(a), fibrinogen and CV event rate, and almost no clinically significant side-effects. PMID- 10809996 TI - Effects of short-term administration of growth hormone in healthy young men, women, and women taking oral contraceptives. AB - OBJECTIVE: Effects of short-term administration of growth hormone (GH) with respect to gender and oral contraceptives (OCs) were investigated in healthy young adults. DESIGN: Open, prospective 2-week study. SETTING: Clinical research centre, university hospital. SUBJECTS: Three groups of healthy young adults were included: six men, six women with normal menstrual cycles, and six women taking OCs. INTERVENTIONS: The subjects were given recombinant human GH subcutaneously for 2 weeks: 1 U m-2 body surface daily during the first week, and 3 U m-2 daily during the second week. MAIN OUTCOME MEASURES: Serum samples were drawn in the morning after overnight fasting on days 0, 3, 7, 10 and 14, and were analysed for GH, insulin-like growth factor 1 (IGF-1), insulin, testosterone, sex hormone binding globulin (SHBG), lipids and markers of bone metabolism. Second-void morning urine was analysed for deoxypyridinoline (Dpyr). RESULTS: During administration of GH, serum IGF-1 increased in the men and in the women without OCs (86 and 52%, respectively). In the OC women, IGF-1 did not change significantly. Serum insulin increased in all three groups, with the largest change (122%) in the men and the smallest (47%) in the OC women. Blood glucose was unchanged. Total cholesterol, low-density lipoprotein (LDL) cholesterol and the LDL/high-density lipoprotein cholesterol ratio were reduced in the men only. Biochemical markers of bone resorption (serum procollagen type I, urinary Dpyr) increased in the men, and markers of bone formation (serum osteocalcin and telopeptide of collagen type I) increased in the men as well as in the women not taking OCs. The testosterone/SHBG ratio increased in the men on account of a reduction in SHBG. CONCLUSION: The response to short-term administration of GH differed in the three groups, with the largest effect in the men and the smallest in the OC women. The inhibitory influence of contraceptives underlines the role of sex steroids in modulating the susceptibility to GH. PMID- 10809997 TI - Marked improvement in survival after acute myocardial infarction in middle-aged men but not in women. The Northern Sweden MONICA study 1985-94. AB - OBJECTIVES: The aim of this study was to analyse time trends in survival after acute myocardial infarction with special emphasis on sex differences. DESIGN: Within the framework of the population-based WHO MONICA Project, all acute myocardial infarction events were recorded in the age group 25-64 years in northern Sweden during the period 1985-94. All first-ever myocardial infarction patients were followed for information on vital status. SUBJECTS: A total of 3397 men and 860 women with acute myocardial infarction, during the period between 1985 and 1994. MAIN OUTCOME MEASURES: Case fatality rates after first-ever acute myocardial infarction. RESULTS: When compared with the 1985-86 cohort, the age adjusted odds ratio for death within 1 year after acute myocardial infarction was 0.59 (95% CI 0.46-0.76) in the 1993-94 male cohort but 0.99 (95% CI 0.61-1.60) in the female 1993-94 cohort. Corresponding age-adjusted proportions of death within 1 year were 33.3% and 22.9% in men and 27.5% and 27.3% in women in 1985-86 and 1993-94, respectively. The odds ratio for 3-year case fatality amongst those who survived the first 28 days was 0.34 (95% CI 0.21-0.55) in 1991-92 compared with 1985-86 in men and 0.91 (0.43-1.94) in women. CONCLUSION: Both short- and long term survival after AMI have improved markedly in men over the last decade. There is a disturbing sex difference in that, during the same period, survival in women with AMI has not improved at all. This sex difference was not explained by differences in conventional prognostic factors. PMID- 10809998 TI - Distribution and determinants of ischaemic heart disease in an urban population. A study from the myocardial infarction register in Malmo, Sweden. AB - OBJECTIVE: Age adjusted incidence of myocardial infarction has been found to vary substantially between the residential areas of the city of Malmo. The objective of this study was to assess the extent to which major biological risk factors and socio-economic circumstances account for the differences in incidence of and mortality from myocardial infarction. DESIGN: Ecological study of risk factor prevalence and incidence and mortality from myocardial infarction. SETTING: Seventeen administrative areas in Malmo, Sweden. SUBJECTS: Assessment of risk factor prevalence was based on 28 466 men and women, ranging from 45 to 73 years old, who were recruited as participants in the Malmo Diet and Cancer study. Information on serum lipids was available in a random subsample of 5362 subjects. Information about socio-economic level of the residential area was based on statistics from the Malmo City Council and Statistics Sweden. MAIN OUTCOME MEASURES: Weighted least square regressions between prevalence of risk factors (i.e. smoking, hypertension, obesity, diabetes, hypercholesterolemia and hypertriglyceridemia), a myocardial infarction risk score, a socio-economic score and incidence and mortality from myocardial infarction. RESULTS: The risk factor prevalence and myocardial infarction incidence was highest in areas with low socio-economic level. Prevalence of smoking, obesity and hypertension was significantly associated with myocardial infarction incidence and mortality rates amongst men (all r > 0.60). Prevalence of smoking was significantly associated with incidence and mortality from myocardial infarction amongst women (r = 0.66 and r = 0.61, respectively). A myocardial infarction risk score based on four biological risk factors explained 40-60% of the intra-urban geographical variation in myocardial infarction incidence and mortality. The socio-economic score added a further 2-16% to the explained variance. CONCLUSION: In an urban population with similar access to medical care, well-known biological cardiovascular risk factors account for a substantial proportion of the intra urban geographical variation of incidence of and mortality from myocardial infarction. The socio-economic circumstances further contribute to the intra urban variation in disease. PMID- 10809999 TI - Reduced thiamine phosphate, but not thiamine diphosphate, in erythrocytes in elderly patients with congestive heart failure treated with furosemide. AB - OBJECTIVES: To measure the concentrations of thiamine and thiamine esters by high pressure liquid chromatography (HPLC) in elderly patients treated with furosemide for heart failure and in a control group. DESIGN: A cross-sectional study of blood thiamine and thiamine ester concentrations. SUBJECTS: Forty-one patients admitted to hospital for heart failure and 34 elderly living at home. No vitamin supplementation was allowed. RESULTS: Compared with the healthy controls, furosemide-treated patients had significantly reduced whole blood thiamine phosphate (TP; 4.4 +/- 2.2 vs. 7.6 +/- 2.0 nmol L-1) and thiamine diphosphate (TPP; 76 +/- 21.5 vs. 91 +/- 19.8 nmol L-1) (mean +/- SD). When the thiamine concentrations were related to the haemoglobin concentrations, which were reduced in the heart failure patients, the levels of TP (nmol g-1 Hb) were 0.38 +/- 0.26 vs. 0.54 +/- 0.17 (P < 0.0001), and of TPP were 6.35 +/- 1.76 vs. 6.37 +/- 1.29 (P = 0.95). There were no differences in T and TP concentrations in plasma between the two groups. CONCLUSIONS: The elderly patients with heart failure treated with furosemide have not reduced the storage form of thiamine, TPP, but only TP. This change is most likely not an expression of a thiamine deficiency, but rather of an altered metabolism of thiamine, which is not understood at present. PMID- 10810000 TI - The risk of ipsilateral versus contralateral recurrent deep vein thrombosis in the leg. The DURAC Trial Study Group. AB - OBJECTIVES: To investigate the risk of ipsilateral versus contralateral recurrent deep vein thrombosis in the leg. DESIGN: An open prospective long term follow-up multicentre trial. Patients were followed by frequent outpatient visits at each centre during the first 12 months after inclusion and thereafter annually. SETTING: Sixteen hospitals in central Sweden. SUBJECTS: A total of 790 consecutive patients with objectively verified first episode of acute deep vein thrombosis and without diagnosed malignant disease were recruited from a randomized study comparing 6 weeks with 6 months of oral antivitamin K therapy as secondary thromboprophylaxis. MAIN OUTCOME MEASURES: Deep vein thrombosis in the contralateral leg was confirmed by venography or ultrasound. With regard to the ipsilateral leg, venography was required. RESULTS: A recurrent episode of venous thromboembolism was documented in 192 patients after a mean (+/-SD) period of 31(+/-29) months. In 26 additional patients with ipsilateral symptoms the diagnostic critera were not fulfilled. One hundred and eleven patients have deceased and 69 patients withdrew from the study. The 392 patients without recurrent episodes were followed for a median of 96 months with 90% for at least 48 months. An objectively verified recurrent contralateral and ipsilateral deep vein thrombosis occurred in 95 and 54 cases, respectively, and in 41 patients pulmonary embolism was documented. In two patients thromboses with unusual locations were registered. The risk of contralateral versus ipsilateral recurrence was significantly increased with a risk ratio of 1.6 (95% confidence interval 1.4-1.9) in a time to event model. In a multivariate analysis none of the investigated variables were significantly associated with the side of recurrent thrombosis. CONCLUSIONS: The risk of a recurrent deep vein thrombosis is increased in the contralateral leg. This brings into question the importance of an impaired venous flow for recurrent episodes of thrombosis. PMID- 10810001 TI - Fat intake and food choices during weight reduction with diet, behavioural modification and a lipase inhibitor. AB - OBJECTIVE: To study the composition of fat intake and fat-rich meals consumed during a trial in which obese subjects were treated with a lipase-inhibitor or placebo, with emphasis on food choices and eating hours. DESIGN: Patients were instructed to record all food and drink taken for four days prior to each dietician visit. The food diaries from all scheduled 15 treatment visits were analysed for nutritional content and composition and for temporal distribution. All meals containing 25 g of fat were defined as fat-rich. SUBJECTS: Twenty-eight women and six men, mean age 45.2 +/- 10.9 (SD) years with a mean body mass index of 37.3 +/- 3.3 (SD) kg m-2 at the beginning of the study. RESULTS: Fat intake, both as absolute weight and as energy % was generally higher in the placebo group but no significant trend over time could be seen. Fat rich meals were increased by 59% towards the end of the study. Most fat rich meals were eaten at lunch and dinner. Cooking fat, fatty sauces, meat dishes and cheese contributed to the major proportion of fat, both for placebo and drug treated subjects. No major changes were seen in food choice over time. CONCLUSION: A lipase inhibitor may affect the amount of fat ingested but does not seem to change major sources of fat. The typical fat-rich meal consumed by these subjects was a meat dish, consumed in the evening. PMID- 10810002 TI - Discrimination of polymorphic forms of a drug product by localized thermal analysis. AB - In chemical processing, it is important to distinguish between and identify polymorphic forms. We demonstrate the novel use of scanning thermal microscopy (SThM) and localized thermal analysis to distinguish and identify polymorphic forms of the drug cimetidine. These forms cannot be resolved by classical bulk thermal analysis. SThM reveals a sample consisting of a 50 : 50 mixture of the polymorphs contains regions of different thermal conductivity, corresponding to the different polymorphs. Localized thermal analysis of small volumes of pure polymorphic samples (approximately 50 microm3) shows that the origin of the thermal conductivity contrast lies, at least in part, with the presence of a surface water layer on the more hydrophilic polymorph. PMID- 10810003 TI - Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. AB - Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide-field fluorescence microscope. The sample is illuminated with a series of excitation light patterns, which cause normally inaccessible high-resolution information to be encoded into the observed image. The recorded images are linearly processed to extract the new information and produce a reconstruction with twice the normal resolution. Unlike confocal microscopy, the resolution improvement is achieved with no need to discard any of the emission light. The method produces images of strikingly increased clarity compared to both conventional and confocal microscopes. PMID- 10810004 TI - Fluorescence photobleaching-based image standardization for fluorescence microscopy AB - A method is presented for the standardization of images acquired with fluorescence microscopy, based on the knowledge of spatial distributions proportional to the microscope's absolute excitation intensity and fluorescence detection efficiency distributions over the image field. These distributions are determined using a thin fluorescent test layer, employed under practically mono exponential photobleaching conditions. It is demonstrated that these distributions can be used for (i) the quantitative evaluation of differences between both the excitation intensity and the fluorescence detection efficiency of different fluorescence microscopes and (ii) the standardization of images acquired with different microscopes, permitting the deduction of quantitative relationships between images obtained under different imaging conditions. PMID- 10810005 TI - Polarization confocal microscopy and congo red fluorescence: a simple and rapid method to determine the mean cellulose fibril orientation in plants. AB - The mean or net preferential orientation of cellulose fibrils in plant cell walls is detected with polarization confocal laser scanning microscopy using the fluorescence dichroism of Congo Red. Single cells, arrays of cells in a tissue, or the epidermis of whole organs can be assayed in vivo. Aerial parts require an extra pectinase treatment because of the cuticle, which is impermeable to aqueous solutions. Peeling off the epidermis can be an elegant alternative, especially for leaves. With this method the net preferential fibril orientation can be related to the symmetry axis of the cell in quantitative terms. Data issuing from this approach are useful in current research on plant biomechanics. PMID- 10810006 TI - Secondary ion mass spectrometry imaging of the fixation of 15N-labelled NO in pollen grains. AB - We used secondary ion mass spectrometry to image cellular targets of nitrogen oxides (widespread air pollutants) in pollen grains of birch (Betula verrucosa Ehrh.) and cockfoot (Dactylis glomerata L.). The pollen samples were exposed to air supplemented with high doses of 15NO. The pollen grains were then fixed, dehydrated using a newly developed 'vapour phase' preparation method and embedded in LRW resin. Semithin sections were then analysed. Imaging was performed in scanning mode. As usual, the two isotopes 14N and 15N were imaged as 12C14N- and 12C15N-, respectively. The isotopic percentages of 15N were quantitatively determined either by image processing or by direct analysis. We show that the preferential areas of NO fixation in the pollen cell are the sporoderm and discrete intracytoplasmic structures that we tentatively describe as globoid-like structures similar to those encountered in seeds. PMID- 10810007 TI - Long freeze-drying times are not necessary during the preparation of thin sections for X-ray microanalysis. AB - The temperature profile that occurs when a brass block warms up in a vacuum evaporation unit was determined. Freshly drawn human blood was concentrated by centrifugation, and the pellet was cryofixed and cryosectioned. The cryosections were subject to different freeze-drying protocols, using a freeze-drier with a temperature-controlled stage, to determine the effect of freeze-drying time on element distribution. Spectra were collected by spot analyses at various distances across the interface between red cells and plasma. Concentrations of sodium were high and variable outside the cell and low in the cell interior, with potassium showing the reverse distribution. The number of counts under the iron peak closely followed the potassium distribution. The concentration of sodium was higher than expected at 40 nm inside the cell membrane. This was attributed to the formation of ice crystals at the interface between the cells and plasma during cryofixation and the use of a wide probe size. PMID- 10810008 TI - Focused ion beam sample preparation of continuous fibre-reinforced ceramic composite specimens for transmission electron microscopy AB - The microanalysis of interfaces in fibre-reinforced composite materials is dependent on the successful preparation of specimens suitable for transmission electron microscope (TEM) inspection. Ideal samples should possess large amounts of structurally intact and uniform thin area in the fibre/matrix interface regions of the samples. Because fibre/matrix interfaces in this class of materials are often designed to fail under mechanical stress, conventionally prepared samples are prone to interfacial failure and differential thinning, both of which preclude detailed TEM microanalysis. These effects were seen in a conventionally dimpled and ion-beam-thinned specimen prepared from a continuous fibre reinforced ceramic composite composed of CaWO4-coated Nextel 610TM fibres in an alumina matrix. The dimpled specimen showed large amounts of interfacial failure, with only thick regions of the specimen left intact. To overcome these limitations, a focused ion beam (FIB) technique was applied to this same material. The superiority of the FIB-produced sample is evident in both the morphology and scanning transmission electron microscopy analyses of the sample. PMID- 10810009 TI - Methodological principles for fractal analysis of trabecular bone. AB - A standardized methodology for the fractal analysis of histological sections of trabecular bone has been established. A modified box counting method has been developed for use on a PC-based image analyser. The effect of image analyser settings, magnification, image orientation and threshold levels was determined. Also, the range of scale over which trabecular bone is effectively fractal was determined and a method formulated to calculate objectively more than one fractal dimension from the modified Richardson plot. The results show that magnification, image orientation and threshold settings have little effect on the estimate of fractal dimension. Trabecular bone has a lower limit below which it is not fractal (lambda < 25 microm) and the upper limit is 4250 microm. There are three distinct fractal dimensions for trabecular bone (sectional fractals), with magnitudes greater than 1.0 and less than 2.0. It has been shown that trabecular bone is effectively fractal over a defined range of scale. Also, within this range, there is more than one fractal dimension, describing spatial structural entities. Fractal analysis is a model-independent method for describing a complex multifaceted structure, which can be adapted for the study of other biological systems. This may be at the cell, tissue or organ level and complements conventional histomorphometric and stereological techniques. PMID- 10810010 TI - Classification of spatial textures in benign and cancerous glandular tissues by stereology and stochastic geometry using artificial neural networks. AB - Stereology and stochastic geometry can be used as auxiliary tools for diagnostic purposes in tumour pathology. Whether first-order parameters or stochastic geometric functions are more important for the classification of the texture of biological tissues is not known. In the present study, volume and surface area per unit reference volume, the pair correlation function and the centred quadratic contact density function of epithelium were estimated in three case series of benign and malignant lesions of glandular tissues. The information provided by the latter functions was summarized by the total absolute areas between the estimated curves and their horizontal reference lines. These areas are considered as indicators of deviation of the tissue texture from a completely uncorrelated volume process and from the Boolean model with convex grains, respectively. We used both areas and the first-order parameters for the classification of cases using artificial neural networks (ANNs). Learning vector quantization and multilayer feedforward networks with backpropagation were applied as neural paradigms. Applications included distinction between mastopathy and mammary cancer (40 cases), between benign prostatic hyperplasia and prostatic cancer (70 cases) and between chronic pancreatitis and pancreatic cancer (60 cases). The same data sets were also classified with linear discriminant analysis. The stereological estimates in combination with ANNs or discriminant analysis provided high accuracy in the classification of individual cases. The question of which category of estimator is the most informative cannot be answered globally, but must be explored empirically for each specific data set. Using learning vector quantization, better results could often be obtained than by multilayer feedforward networks with backpropagation. PMID- 10810011 TI - Adverse reactions to vaccines in the tropics. PMID- 10810013 TI - A prospective evaluation of a clinical algorithm for the diagnosis of malaria in Gambian children. AB - Diagnosis of clinical malaria remains difficult, especially in areas where a high proportion of the asymptomatic population have parasitaemia, for the symptoms and signs of malaria overlap with those of other common childhood diseases, such as acute lower respiratory tract infections. However, a study of symptoms and signs in a group of children who presented to Farafenni Health Centre, The Gambia with a history of recent fever identified a group of signs and symptoms which were strong predictors of malaria as opposed to other febrile illnesses. Using these predictors, an algorithm was developed which could be used by fieldworkers and which had a similar sensitivity and specificity for the diagnosis of malaria as that of an experienced paediatrician working without laboratory support. This algorithm has been validated prospectively on 518 children who presented to the Medical Research Council clinic at Basse, The Gambia with fever or a history of recent fever during a 10-month period. A fieldworker obtained a detailed history from the parent or guardian of each child and performed a clinical examination which included measurement of axillary temperature and respiratory rate. Packed cell volume was measured and a thick smear was examined for malaria parasites. A malaria score, based on the presence or absence of malaria-related signs and symptoms, was determined for 382 children who were seen at the clinic during the high transmission season. Using the cut-off score which was optimal during the previous retrospective study, a sensitivity of 70% and a specificity of 77% for a diagnosis of malaria was obtained. The optimal cut-off score for the Basse population was a score of 7; this gave a sensitivity of 88% and a specificity of 62%, figures comparable to those obtained by an experienced paediatrician without laboratory support. PMID- 10810015 TI - Economic evaluation of parasitic diseases: a critique of the internal and external validity of published studies. AB - It was estimated that in 1990, major parasitic diseases accounted for 11.7% of the disease burden from communicable disease. As advances in the prevention, diagnosis and treatment of parasitic diseases are made and implemented, there is a growing economic literature to help decision-makers choose the most efficient control method. The aim of this paper is to identify, describe and analyse the available published data on the efficiency of control strategies against parasitic diseases. Internal validity is assessed through the quality of economic evaluations over time using a series of standard questions, and external validity is assessed in terms of the potential to extrapolate results to other settings. This leads to a discussion of the legitimacy and feasibility of pooling data or results from studies for priority setting in the health sector, resulting in three recommendations: to increase the coverage of economic evaluations for parasitic diseases and types of interventions; to improve the internal validity of studies through guidelines and review procedures; and to explore the external validity of research results by examining their predictive validity across settings. PMID- 10810018 TI - Single small enhancing CT lesion in thai patients with acute symptomatic seizures: a clinico-radiological study AB - objective To identify the frequency of single small enhancing CT lesion cases (SSECTL) in a provincial hospital in Thailand and verify a set of clinico radiological criteria for the diagnosis of benign SSECTL. methods All patients who fulfilled the following criteria were recruited: acute symptomatic focal seizures with or without secondary generalization; minimal or no neurological deficit; no evidence of raised intracranial pressure; no evidence of systemic disease; CT scan showing SSECTL of < 20 mm diameter. After recruitment, patients received antiepileptic drugs and other symptomatic treatment. A CT scan of the brain was repeated every 2 weeks until the lesion had significantly resolved, which was defined as a 50% decrease in size. A CT scan was then taken every 4 weeks until the lesion had completely disappeared or turned into a calcified spot. The data from all other patients presenting with seizures and solitary enhancing lesion on the CT scan who did not fulfil the inclusion criteria were also reviewed. results 972 patients with seizure disorder were recruited. 110 patients (11.3%) presented with seizure and solitary enhancing lesion on the CT scan, 22 of whom (20%) fulfilled the inclusion criteria. 20 of the 22 patients had SSECTL with a spontaneous resolution: 14 (70%) within 4 weeks and 18 (90%) within 8 weeks. Two patients had a progressive course. One patient with a solitary enhancing CT lesion > 20 mm had spontaneous resolution. 87 patients had another diagnoses. The diagnostic criteria for benign SSECTL were 95.23% sensitive, 97.75% specific, had a positive predictive value of 90.91% and a negative predictive value of 98.86%. conclusion SSECTL is not uncommon in our provincial hospital. The clinico-radiological criteria proposed by Rajshekhar (1991) are valid and reliable in predicting a benign outcome. CT follow-up within the first 4 weeks is critical. PMID- 10810017 TI - Single small enhancing CT lesion in Thai patients with acute symptomatic seizures: a clinico-radiological study. AB - OBJECTIVE: To identify the frequency of single small enhancing CT lesion cases (SSECTL) in a provincial hospital in Thailand and verify a set of clinico radiological criteria for the diagnosis of benign SSECTL. METHODS: All patients who fulfilled the following criteria were recruited: acute symptomatic focal seizures with or without secondary generalization; minimal or no neurological deficit; no evidence of raised intracranial pressure; no evidence of systemic disease; CT scan showing SSECTL of < 20 mm diameter. After recruitment, patients received antiepileptic drugs and other symptomatic treatment. A CT scan of the brain was repeated every 2 weeks until the lesion had significantly resolved, which was defined as a 50% decrease in size. A CT scan was then taken every 4 weeks until the lesion had completely disappeared or turned into a calcified spot. The data from all other patients presenting with seizures and solitary enhancing lesion on the CT scan who did not fulfil the inclusion criteria were also reviewed. RESULTS: 972 patients with seizure disorder were recruited. 110 patients (11.3%) presented with seizure and solitary enhancing lesion on the CT scan, 22 of whom (20%) fulfilled the inclusion criteria. 20 of the 22 patients had SSECTL with a spontaneous resolution: 14 (70%) within 4 weeks and 18 (90%) within 8 weeks. Two patients had a progressive course. One patient with a solitary enhancing CT lesion > 20 mm had spontaneous resolution. 87 patients had another diagnoses. The diagnostic criteria for benign SSECTL were 95.23% sensitive, 97.75% specific, had a positive predictive value of 90.91% and a negative predictive value of 98.86%. CONCLUSION: SSECTL is not uncommon in our provincial hospital. The clinico-radiological criteria proposed by Rajshekhar (1991) are valid and reliable in predicting a benign outcome. CT follow-up within the first 4 weeks is critical. PMID- 10810019 TI - Markers of inflammation in children with severe malarial anaemia. AB - OBJECTIVE: To investigate if severe malarial anaemia is associated with a specific immune response pattern, we determined serum levels of neopterin (a marker of activation of macrophages by interferon-gamma) and of the anti inflammatory cytokines, interleukins 4 and 10. METHODS: Zambian children < 6 years of age presenting to a rural hospital with cerebral malaria were studied. Twenty-one children with admission haemoglobin concentrations /= 7 g/dl served as a control group. RESULTS: Logistic regression modelling indicated that a 10-fold rise in serum neopterin concentrations was associated with a 50 fold increase in the estimated odds of having severe anaemia (P = 0.015), while a 10-fold rise in serum interleukin 4 concentrations was associated with a 10-fold decrease in the estimated odds of having severe anaemia (P = 0.023). Increasing serum interleukin 10 concentrations, measured in less than half of the subjects, were associated with a nonsignificant reduction in the odds of having severe anaemia (P = 0.095). CONCLUSION: Development of severe malarial anaemia may be directly associated with serum neopterin concentrations and inversely correlated with serum interleukin 4 levels. PMID- 10810020 TI - Markers of inflammation in children with severe malarial anaemia AB - objective To investigate if severe malarial anaemia is associated with a specific immune response pattern, we determined serum levels of neopterin (a marker of activation of macrophages by interferon-gamma) and of the anti-inflammatory cytokines, interleukins 4 and 10. methods Zambian children < 6 years of age presenting to a rural hospital with cerebral malaria were studied. Twenty-one children with admission haemoglobin concentrations /= 7 g/dl served as a control group. results Logistic regression modelling indicated that a 10-fold rise in serum neopterin concentrations was associated with a 50-fold increase in the estimated odds of having severe anaemia (P = 0.015), while a 10-fold rise in serum interleukin 4 concentrations was associated with a 10-fold decrease in the estimated odds of having severe anaemia (P = 0.023). Increasing serum interleukin 10 concentrations, measured in less than half of the subjects, were associated with a nonsignificant reduction in the odds of having severe anaemia (P = 0.095). conclusion Development of severe malarial anaemia may be directly associated with serum neopterin concentrations and inversely correlated with serum interleukin 4 levels. PMID- 10810021 TI - Land use change alters malaria transmission parameters by modifying temperature in a highland area of Uganda. AB - As highland regions of Africa historically have been considered free of malaria, recent epidemics in these areas have raised concerns that high elevation malaria transmission may be increasing. Hypotheses about the reasons for this include changes in climate, land use and demographic patterns. We investigated the effect of land use change on malaria transmission in the south-western highlands of Uganda. From December 1997 to July 1998, we compared mosquito density, biting rates, sporozoite rates and entomological inoculation rates between 8 villages located along natural papyrus swamps and 8 villages located along swamps that have been drained and cultivated. Since vegetation changes affect evapotranspiration patterns and, thus, local climate, we also investigated differences in temperature, humidity and saturation deficit between natural and cultivated swamps. We found that on average all malaria indices were higher near cultivated swamps, although differences between cultivated and natural swamps were not statistically significant. However, maximum and minimum temperature were significantly higher in communities bordering cultivated swamps. In multivariate analysis using a generalized estimating equation approach to Poisson regression, the average minimum temperature of a village was significantly associated with the number of Anopheles gambiae s.l. per house after adjustment for potential confounding variables. It appears that replacement of natural swamp vegetation with agricultural crops has led to increased temperatures, which may be responsible for elevated malaria transmission risk in cultivated areas. PMID- 10810023 TI - Targeting of Wolbachia endobacteria in Litomosoides sigmodontis: comparison of tetracyclines with chloramphenicol, macrolides and ciprofloxacin. AB - Endobacteria of the genus Wolbachia in filarial nematodes are related to Rickettsiaceae and can be depleted by tetracycline antibiotics. This depletion blocks female worm development as well as early embryogenesis, in contrast to the currently used microfilaricidal ivermectin which blocks only the last stage of embryogenesis. Since targeting Wolbachia is becoming an area of research for the treatment of human filariases, it was investigated if antibiotics other than tetracyclines are able to deplete Wolbachia from filariae. BALB/c mice infected with the rodent filaria Litomosoides sigmodontis were treated with erythromycin, chloramphenicol or ciprofloxacin. All drugs were well resorbed and resulted in serum levels clearly above breakpoint levels for bacteria susceptible to the respective antibiotic. However, contrary to tetracycline, none of these antibiotics depleted Wolbachia or altered worm development and fertility, as evidenced by immunohistology, immunoelectron microscopy and semiquantitative PCR. PMID- 10810024 TI - Targeting of wolbachia endobacteria in litomosoides sigmodontis: comparison of tetracyclines with chloramphenicol, macrolides and ciprofloxacin AB - Endobacteria of the genus Wolbachia in filarial nematodes are related to Rickettsiaceae and can be depleted by tetracycline antibiotics. This depletion blocks female worm development as well as early embryogenesis, in contrast to the currently used microfilaricidal ivermectin which blocks only the last stage of embryogenesis. Since targeting Wolbachia is becoming an area of research for the treatment of human filariases, it was investigated if antibiotics other than tetracyclines are able to deplete Wolbachia from filariae. BALB/c mice infected with the rodent filaria Litomosoides sigmodontis were treated with erythromycin, chloramphenicol or ciprofloxacin. All drugs were well resorbed and resulted in serum levels clearly above breakpoint levels for bacteria susceptible to the respective antibiotic. However, contrary to tetracycline, none of these antibiotics depleted Wolbachia or altered worm development and fertility, as evidenced by immunohistology, immunoelectron microscopy and semiquantitative PCR. PMID- 10810025 TI - Microheterogeneity of serogroup A (subgroup III) Neisseria meningitidis during an outbreak in northern Ghana. AB - During a meningitis outbreak in the eastern subdistrict of the Kassena-Nankana District of the Upper East Region of Ghana, we analysed cerebrospinal fluid from suspected meningitis cases for the most common causative organisms. In 50 of 92 samples analysed, serogroup A Neisseria meningitidis were detected. The ages of serogroup A N. meningitidis patients ranged from 4 months to 64 years. The case fatality ratio was 20%. Coma or stupor on presentation worsened the prognosis. All serogroup A N. meningitidis isolates recovered revealed the A: 4: P1.9, 20 phenotype characteristic for the subgroup III clonal grouping. No evidence for resistance to penicillin G, chloramphenicol, cefotaxime, ciprofloxacin, rifampicin or tetracycline was found. All strains were resistant to sulphadiazine. Restriction analysis patterns of opa, iga and ingA genes were characteristic for the majority of N. meningitidis serogroup A subgroup III bacteria isolated in Africa after the 1987 epidemic in Mecca. Differences in pulsed-field gel electrophoresis patterns of NheI and SpeI digested DNA revealed microheterogeneity among the Ghanaian isolates. PMID- 10810026 TI - Microheterogeneity of serogroup A (subgroup III) neisseria meningitidis during an outbreak in northern ghana AB - During a meningitis outbreak in the eastern subdistrict of the Kassena-Nankana District of the Upper East Region of Ghana, we analysed cerebrospinal fluid from suspected meningitis cases for the most common causative organisms. In 50 of 92 samples analysed, serogroup A Neisseria meningitidis were detected. The ages of serogroup A N. meningitidis patients ranged from 4 months to 64 years. The case fatality ratio was 20%. Coma or stupor on presentation worsened the prognosis. All serogroup A N. meningitidis isolates recovered revealed the A: 4: P1.9, 20 phenotype characteristic for the subgroup III clonal grouping. No evidence for resistance to penicillin G, chloramphenicol, cefotaxime, ciprofloxacin, rifampicin or tetracycline was found. All strains were resistant to sulphadiazine. Restriction analysis patterns of opa, iga and ingA genes were characteristic for the majority of N. meningitidis serogroup A subgroup III bacteria isolated in Africa after the 1987 epidemic in Mecca. Differences in pulsed-field gel electrophoresis patterns of NheI and SpeI digested DNA revealed microheterogeneity among the Ghanaian isolates. PMID- 10810027 TI - Coping strategies of health personnel during economic crisis: A case study from Cameroon. AB - OBJECTIVES: Severe economic crisis compelled many governments in Sub-Saharan Africa to adopt structural adjustment programmes. This was accompanied by price increases and cuts in the salaries of civil servants. We explored how health personnel in one province of Cameroon coped with this situation, and what the perceived effects on service quality were. METHODS: Key informant and focus group interviews with government and mission (church) health personnel; interviews with service users to validate the findings. RESULTS: Government health personnel had experienced larger cuts in salaries than their mission counterparts; they no longer received allowances and incentives still available to mission personnel and appeared more demotivated. Most government and mission personnel reported legal after-hours income raising activities. Government personnel frequently reported additional 'survival strategies' such as parallel selling of drugs, requesting extra charges for services, and running private practices during work hours. There was a high level of self criticism among government personnel indicating a dissonance between their attitude and practices. They considered these practices negative and harmful for service users. CONCLUSION: Remedial action is urgent. Options include reinstating allowances for good performance and ensuring regular supervision without blaming individual health workers for problems caused by the state of the health system. PMID- 10810028 TI - Coping strategies of health personnel during economic crisis: A case study from cameroon AB - objectives Severe economic crisis compelled many governments in Sub-Saharan Africa to adopt structural adjustment programmes. This was accompanied by price increases and cuts in the salaries of civil servants. We explored how health personnel in one province of Cameroon coped with this situation, and what the perceived effects on service quality were. methods Key informant and focus group interviews with government and mission (church) health personnel; interviews with service users to validate the findings. results Government health personnel had experienced larger cuts in salaries than their mission counterparts; they no longer received allowances and incentives still available to mission personnel and appeared more demotivated. Most government and mission personnel reported legal after-hours income raising activities. Government personnel frequently reported additional 'survival strategies' such as parallel selling of drugs, requesting extra charges for services, and running private practices during work hours. There was a high level of self criticism among government personnel indicating a dissonance between their attitude and practices. They considered these practices negative and harmful for service users. conclusion Remedial action is urgent. Options include reinstating allowances for good performance and ensuring regular supervision without blaming individual health workers for problems caused by the state of the health system. PMID- 10810029 TI - Congenital transmission of Trypanosoma cruzi: an operational outline for detecting and treating infected infants in north-western Argentina. AB - We designed a set of procedures for first-line local health services to detect and treat the congenital transmission of Trypanosoma cruzi at a province-wide scale, and field-tested the programme in the province of Tucuman, northwestern Argentina, from 1992 to 1994. The programme consists of routine screening of pregnant women for seroreactivity to T. cruzi, serological and parasitological follow-up of the newborn at least twice during the first year of age, treatment of the infected infants, and evaluation of the outcome. 927 (5.5%) of 16 842 pregnant women were seroreactive to T. cruzi by indirect haemagglutination assay and ELISA. Twenty-one (6.7%) of 315 newborns to seroreactive mothers were diagnosed as infected with T. cruzi parasites microhaematocrit concentration before 30 days of age. Five newborns who initially tested negative had a T. cruzi infection detected by microhaematocrit and/or serological techniques at 3 or 6 months of age. Thus, congenital infection was diagnosed in 26 (7.1%) infants born to seroreactive women and residing in houses free of triatomine bugs. Four of 6 infants born to seroreactive mothers died during the first year of age and had some evidence of T. cruzi infection; one of the deaths was attributed to T. cruzi based on clinical evidence. After specific treatment with nifurtimox or benznidazole, 30 of 32 infants remained parasitologically and serologically negative. This study shows the feasibility of controlling the incidence of congenitally acquired T. cruzi infections at a province-wide scale by means of a specific screening programme at first-line health services level. PMID- 10810031 TI - Prevalence and distribution of soil-transmitted helminth (STH) infections in urban and indigenous schoolchildren in Ortigueira, State of Parana, Brasil: implications for control. AB - Soil-transmitted helminth (STH) infections represent a major public health problem in poor and developing countries. During the period September-October 1998 we conducted an epidemiological survey of STH infections in schoolchildren of an urban area (group A) and an indigenous reserve (group B), in the Municipality of Ortigueira, State of Parana, Brazil, to assess potential benefits of mass treatment. Stool samples were examined for helminth eggs by quantitative (Kato-Katz) technique to determine the prevalence and intensity of intestinal parasitic infection. Moreover, we examined the relationship between prevalence and intensity of STH infections and housing/hygienic factors (by means of a 7 item questionnaire). 236 schoolchildren aged 5-15 years were enrolled, 136 in group A and 100 in group B. The prevalence of STH infections was significantly higher in group B (93%) than in group A (22%) (P < 0.001). Detected parasites were: A. lumbricoides (16.1% prevalence in group A, 88% in group B, P < 0.001), hookworms (5.8% in group A, 52% in group B, P < 0.001) and T. trichiura (5.1% in group A, 2% in group B, P = 0.2). Heavy infections were detected in 2.9% and 23% of the children in group A and B, respectively (P < 0.001). Housing/hygienic indicators were significantly poorer in group B. A statistically significant correlation was observed between total prevalence of STH infections and prevalence of high-intensity infections with most housing/hygienic variables. On the basis of these results, mass treatment and educational interventions were suggested for the indigenous community, whereas target treatment and educational interventions were suggested for the urban community. Even in a geographically homogeneous area different epidemiological realities can be found, which in turn can influence infection levels and control programmes. PMID- 10810033 TI - Animal models in wound healing research: do we need more? PMID- 10810034 TI - Models for use in wound healing research: a survey focusing on in vitro and in vivo adult soft tissue. AB - Many different factors must be considered before selecting a wound healing model to use for a specific study. A wide variety of models have been developed that examine different aspects of the repair response, both in vitro and in vivo. In this review article, we focus on those systems that are most widely used for studies on adult soft tissue healing. Advantages and disadvantages of each are discussed, along with relevant background information to help guide decision making. PMID- 10810035 TI - Oxandrolone, an anabolic steroid, enhances the healing of a cutaneous wound in the rat. AB - The effect of the anabolic steroid oxandrolone on the healing rate of a standardized full thickness linear wound on the back of the rat was studied. Oxandrolone was given orally by gavage in peanut oil at a dose of 0.1 mg/kg/day. A placebo powder in peanut oil was given at the same dose to a control group. Parameters monitored were time to complete wound closure, wound hydroxyproline content and tensile strength, as well as histology. We found that wounds closed completely in 12+/-3 days with oxandrolone, compared to 18+/-3 days for a placebo, a statistically significant difference. The rate of body weight gain was identical in both groups. Hydroxyproline content of the healed incision site was 23+/-4 mg/g tissue vs. 17+/-3 mg/g tissue, while the tensile strength increased to 185+/-13 g/mm2 vs. 102+/-18 g/mm2 in the oxandrolone and placebo groups, respectively. Both parameters were significantly increased with the anabolic steroid. Histologic examination showed a wound that contained more mature and densely packed collagen and was also hypercellular with oxandrolone treatment. We conclude that the anabolic steroid oxandrolone significantly enhanced wound healing unrelated to any generalized increase in protein mass as would be reflected in body weight. PMID- 10810036 TI - Growth hormone does not attenuate the inhibitory effects of sepsis on wound healing. AB - Chronic abdominal sepsis is associated with impaired tissue repair. Treatment of burn patients with growth hormone results in improved healing of skin graft donor sites. The goal of this study was to determine whether administration of growth hormone could attenuate the inhibitory effects of sepsis on cutaneous wound healing. Four groups of male Sprague Dawley rats were studied: control, control + growth hormone, sepsis, and sepsis + growth hormone. Sepsis was caused by implantation of a bacterial focus in the peritoneal cavity. Control animals underwent sham laparotomy, and polyvinyl alcohol sponge implants were placed in subdermal pockets in all animals. Saline or growth hormone (400 microg) was injected subcutaneously every 12 hours. On day 5, the incisional wounds and polyvinyl alcohol sponge implants were harvested. The breaking strength of abdominal incisions was measured. Granulation tissue penetration and quality were determined by scoring polyvinyl alcohol sponge implant histology from 1 to 4 in a blinded fashion. Collagen deposition in polyvinyl alcohol sponge implants was quantitated by hydroxyproline assay. Septic mortality was not altered by growth hormone administration. Septic animals showed a reduction in food consumption for 2 days after surgery (p < 0.05 vs. controls), which was not affected by growth hormone administration. The breaking strength of incisional wounds and hydroxyproline content of polyvinyl alcohol sponge implants was reduced in septic rats (p < 0.001 vs. controls) but administration of growth hormone for 5 days did not improve breaking strength or collagen deposition in either group. We conclude that the administration of growth hormone for 5 days did not improve collagen deposition or breaking strength in cutaneous wounds from control or septic animals. The results suggest that growth hormone treatment is unlikely to improve tissue repair in sepsis-induced catabolic illness. PMID- 10810037 TI - Apoptosis in regenerating and denervated, nonregenerating urodele forelimbs. AB - Denervated limbs of larval salamanders fail to regenerate if amputated and, unlike adult limbs, undergo regression. The cellular basis of the tissue loss is poorly understood. We used TUNEL staining of larval axolotl limbs fixed and sectioned at intervals after bilateral amputation and unilateral denervation to investigate the role of apoptosis during normal limb regeneration and denervated limb regression. In the first week after amputation a small percentage of apoptotic cells was found in both innervated and denervated limbs. During the second week the apoptotic index remained low in the mitotically active mesenchymal cells of the regenerating limbs, but increased twofold in the nondividing, dedifferentiated cells of the regressing limbs. TUNEL-positive cells resembling apoptotic bodies were restricted primarily to the dedifferentiated area beneath the wound epithelium, but were also present within the wound epithelium itself. Macrophages were identified immunohistochemically and were also found in increased numbers in distal areas of the denervated regressing limbs. The results suggest a role for apoptosis in the early phase of normal regeneration and indicate that denervated limb regression involves an increased rate of apoptosis and removal of apoptotic bodies by macrophages and the wound epithelium. PMID- 10810038 TI - Boosting epidermal growth factor receptor expression by gene gun transfection stimulates epidermal growth in vivo. AB - Expression constructs encoding a full-length cDNA encoding the human epidermal growth factor receptor, or reporter gene for green fluorescent protein or luciferase were coated onto gold particles and driven into porcine skin using a gene gun delivery system. Strategies for epidermal growth factor receptor boosting were tested in two types of wounds. For grafted wounds, intact porcine skin was pretreated by the introduction of the epidermal growth factor receptor expression construct 24 hours before its harvesting as a split-thickness skin graft. Partial-thickness excisional wound beds (donor sites) were transfected at the time of their creation. Wound healing parameters were subsequently tested in the presence or absence of excess epidermal growth factor ligand. Initial distributions of gene gun delivered gold particles as well as luciferase expression levels suggested that optimal skin penetrations and expression levels were achieved at 500 psi for intact epidermis and 300 psi for exposed wound beds. At 2 days after gene delivery, visualization of green fluorescent protein by fluorescence microscopy showed focal expression of green fluorescent protein at the advancing epithelial outgrowths found at wound edges or surviving epithelial remnants. Green fluorescent protein expression appeared transient since no green fluorescent protein was noted in specimens removed at 4 days after injury. Northern blot analysis on mRNA isolated from wounds 2 days after introduction of epidermal growth factor receptor coated gold particles by gene gun confirmed the expression of the human epidermal growth factor receptor transgene in both skin grafts and excisional wounds. Skin grafts showed subsequent biological responses to the introduction of excessive epidermal growth factor receptor as well as expression of the human epidermal growth factor receptor construct within healing epidermis. While control autografts (reporter gene treated, epidermal growth factor alone, placebo formula, no treatment) showed few 5'-bromodeoxyuridine labeled cells, epidermal growth factor receptor autografts showed 5' bromodeoxyuridine labeling of nearly every basal cell. Favorable wound healing outcomes were also shown within excisional wounds following in vivo boosting of epidermal growth factor receptor. Four days after receiving epidermal growth factor receptor particle growth factor receptor transgene. Application of topical epidermal growth factor ligand resulted in the highest percentage of resurfacing. Maximal re-epithelialization was noted in wound beds receiving both receptor boosting and excessive daily epidermal growth factor ligand. A modest increase in the thickness of the granulation tissue followed gene therapy with epidermal growth factor receptor. In summary these in vivo data suggest that it is possible to boost in vivo expression of a tyrosine kinase receptor during wound repair. Increased epidermal growth factor receptor expression has an integral impact on cell proliferation, rates of resurfacing and dermal components and merits consideration as a possible therapeutic agent. PMID- 10810039 TI - Hypertrophic scar tissues and fibroblasts produce more transforming growth factor beta1 mRNA and protein than normal skin and cells. AB - Transforming growth factor-beta1 is a well-known fibrogenic cytokine produced by many types of cells including dermal fibroblasts. To investigate whether this fibrogenic cytokine is involved in development of hypertrophic scar, transforming growth factor-beta1 gene expression was evaluated in small skin samples. Because a sufficient quantity of normal skin from patients with hypertrophic scar is not readily available, a reverse transcription-polymerase chain reaction technique was used. Quantitation of gene expression by reverse transcription-polymerase chain reaction is difficult partly due to the lack of suitable complementary RNA standards. We have established a convenient, reliable procedure to construct an internal standard for transforming growth factor-beta1 starting with a gene specific polymerase chain reaction product. After digestion of the polymerase chain reaction product with endonuclease, a small piece of cDNA from human procollagen alpha1(I) cDNA with compatible ends was inserted into the polymerase chain reaction-DNA fragment. The recombinant cDNA was re-amplified by polymerase chain reaction and subcloned into a plasmid containing bacteriophage T7 and T3 promoters. Complementary RNA was prepared from the recombinant plasmid and amplified by reverse transcription-polymerase chain reaction together with the tissue or cellular RNA. After amplification, the products were electrophoresed in an agarose gel containing ethidium bromide. The bands for internal standard and transforming growth factor-beta1 mRNA were scanned, digitized, and plotted against the amount of internal standard complementary RNA added in the reverse transcription-polymerase chain reaction. The number of mRNA molecules/cell was calculated. We examined the transforming growth factor-beta1 mRNA in hypertrophic scar tissue and in normal skin and found that hypertrophic scar tissues expressed five-fold more transforming growth factor-beta1 mRNA than normal skin per unit of wet weight. We used this procedure to quantitate transforming growth factor-beta1 mRNA expression in 5 pairs of fibroblast cultures derived from hypertrophic scar and normal skin. The results showed that hypertrophic scar fibroblast cultures contain significantly more molecules of mRNA for transforming growth factor-beta1 than normal cells (116 +/- 6 vs. 97 +/- 7, p = 0.017, n = 5). These results were supported by Northern analysis for transforming growth factor-beta1 mRNA in the cells and enzyme-linked immunosorbent assay for TGF-beta1 protein in fibroblast conditioned medium. In conclusion, hypertrophic scar tissue and fibroblasts produce more mRNA and protein for transforming growth factor-beta1, which may be important in hypertrophic scar formation. The construction of the gene specific internal standard for reverse transcription-polymerase chain reaction is a simple and reliable procedure useful to quantitate gene expression in a small amount of tissue or number of cells. PMID- 10810040 TI - Stimulation of fibroblast growth in vitro by intermittent radiant warming. AB - A number of clinical studies have suggested that radiant heat improves the healing of selected acute and chronic wounds. The purpose of this study was to investigate in vitro the effect of intermittent radiant heating on the growth of human skin fibroblasts using a radiant heat-producing dressing with a designated temperature of 38 degrees C. In initial experiments cells were seeded in six well plates, maintained in culture at 33-34 degrees C, and warmed daily for three cycles of 1 hour with 1.5 hour intervals. Changes in cell growth and metabolism were determined in sets of triplicate wells by cell counts and a colorimetric assay before and after one week's treatment. After eight days the number of cells in the radiant heat-treated group was 30% higher and the metabolic activity 47%- 90% higher than in the control group. In quiescent fibroblasts which had been maintained for four weeks in low-serum medium, the warming regime completely prevented the decrease in cell number observed in control cells. Our findings suggest that the stimulation of cell proliferation induced by intermittent heating in vitro may indicate a possible mechanism contributing to in vivo effects. PMID- 10810041 TI - Healing of defects in canine articular cartilage: distribution of nonvascular alpha-smooth muscle actin-containing cells. AB - The objective of this study was to evaluate the types of tissue resulting from spontaneous healing of surgically created defects in adult canine articular cartilage up to 29 weeks postoperatively, with specific attention directed toward the presence and distribution of cells containing the contractile actin isoform, alpha-smooth muscle actin. Two 4-mm diameter defects were made in the trochlear groove to the depth of the tidemark in 20 adult mongrel dogs. The areal percentage of specific tissue types in the reparative material was determined histomorphometrically. Immunohistochemistry was employed to evaluate the percentage of alpha-smooth muscle actin-containing cells. The results showed that approximately 50% of the chondrocytes in the superficial zone of the uninvolved articular cartilage expressed alpha-smooth muscle actin. A significantly lower percentage of alpha-smooth muscle actin-positive chondrocytes appeared in the uninvolved deep zone. Notably, the deep zone adjacent to the defect contained a greater percentage of such cells than in the uninvolved deep zone. Also of interest was that a greater percentage of nonvascular cells in the hyaline cartilage and fibrocartilage of the reparative tissue contained alpha-smooth muscle actin-positive cells, compared to the fibrous tissue in the defects. The findings of this study revealed that canine articular cartilage has some potential for spontaneous regeneration, including integration with the calcified cartilage zone. By 29 weeks, up to 40% of an areal cross section of an untreated full-thickness chondral defect was found to fill with hyaline cartilage, with up to 19% judged histologically similar to articular cartilage. The results warrant further consideration of the role of alpha-smooth muscle actin in chondrocytes in normal articular cartilage and in reparative tissue. PMID- 10810042 TI - Atrial fibrillation: part II. PMID- 10810043 TI - Ginkgo biloba and peripheral artery disease. PMID- 10810045 TI - Cholesterol's effects evident in children. PMID- 10810044 TI - Homocysteine a strong risk factor in diabetics. PMID- 10810046 TI - Ask the doctor. I suffer from shortness of breath. My internist has sent me to a cardiologist, who said the cause is not my heart. Then I went to a lung specialist, who said it was not my lungs. I feel like no one wants to take responsibility for helping me! PMID- 10810047 TI - Consumer advice. Buying drugs online. PMID- 10810048 TI - Alternative medicine. Study finds glucosamine doesn't work. PMID- 10810049 TI - Hormone replacement therapy. More risk than benefit? PMID- 10810050 TI - When health care gets overwhelming. PMID- 10810051 TI - Irritable bowel syndrome. New treatment drug on the market. PMID- 10810052 TI - By the way, doctor...I've had small amounts of blood in my bowel movements from time to time for years. My doctor has done a pretty thorough evaluation and has ruled out colon cancer or anything serious: what I have is plain old hemorrhoids. I've heard of an operation using rubber bands that can get rid of them. Should I go ahead and have this procedure done? PMID- 10810053 TI - Treatment of alcoholism -- part II. PMID- 10810054 TI - Evolution and brain aging. PMID- 10810055 TI - Testing child psychotherapy. PMID- 10810056 TI - Forum. Should the expression of emotional memories be a goal of therapy? PMID- 10810058 TI - Priapism and untimely ejaculation: erections that linger and those that don't. PMID- 10810057 TI - Adverse drug reactions: when medicines miss their mark. PMID- 10810059 TI - On call. I am a 69-year-old man who had a radical prostatectomy three years ago. I feel fine and my PSA is undetectable. I've heard that a special scan can detect prostate cancer, but my hospital doesn't seem to know about it. Should I try to arrange one? PMID- 10810060 TI - Estrogen for heart disease: risk or benefit? PMID- 10810061 TI - Exercise: hitting the trail in good form. PMID- 10810062 TI - Breast imaging: mammograms still rule. PMID- 10810063 TI - Fear of falling. PMID- 10810064 TI - By the way, doctor. My surgeon advised me to stop taking gingko biloba before my hip surgery. Can you explain why? Are there any other herbs I should avoid? PMID- 10810066 TI - Ask the doctor. I am generally in good health, but recently had several days of intense, sharp chest pain that got worse when I took a deep breath. My physician told me that I had "pericarditis," and that I shouldn't worry. Should I get any treatment for this problem? PMID- 10810065 TI - More questions on estrogens. PMID- 10810067 TI - Diuretics vs. alpha blockers. PMID- 10810068 TI - By the way, doctor...I am 72 years old and generally healthy, but I have an annoying problem: at the end of the day, my voice gets weak and hoarse. I wonder if it's related to my heartburn, which wakes me up occasionally with an acid taste in my throat and mouth. I drink a lot of coffee, and I used to be a heavy smoker but gave that up 15 years ago. PMID- 10810069 TI - On call. I always undress before I weigh myself at home, but when my doctor weighs me, he doesn't even let me take off my shoes. Wouldn't it be better if I undress first? PMID- 10810071 TI - In brief: A diagnosis is just the beginning PMID- 10810070 TI - Cutting edge: a pill for cancer? PMID- 10810072 TI - By the way, doctor. My doctor recently prescribed Zocor to treat my high cholesterol. She plans to test my liver for possible side effects, which makes me nervous. Is this a dangerous drug? What are its side effects? PMID- 10810073 TI - Clarifying question meaning in a household telephone survey. AB - This study contrasts two interviewing techniques that reflect different tacit assumptions about communication. In one, strictly standardized interviewing, interviewers leave the interpretation of questions up to respondents. In the other, conversational interviewing, interviewers say whatever it takes to make sure that questions are interpreted uniformly and as intended. Respondents from a national sample were interviewed twice. Each time they were asked the same factual questions from ongoing government surveys, five about housing and five about recent purchases. The first interview was strictly standardized; the second was standardized for half the respondents and conversational for the others. Respondents in a second conversational interview answered differently than in the first interview more often, and for reasons that conformed more closely to official definitions, than respondents in a second standardized interview. This suggests that conversational interviewing improved comprehension, although it also lengthened interviews. We conclude that respondents in a national sample may misinterpret certain questions frequently enough to compromise data quality and that such misunderstandings cannot easily be eliminated by pretesting and rewording questions alone. More standardized comprehension may require less standardized interviewer behavior. PMID- 10810074 TI - Rocking the Vote: Using Personalized Messages to Motivate Voting among Young Adults. AB - We examined a nationwide effort to encourage young adults to vote in the 1996 U.S. presidential election. During the year before the election, individuals were given the chance to sign and self-address one of two kinds of postcards pledging to vote; these cards were mailed back to the individuals within 2 weeks prior to the election. It is important to note that some individuals completed pledge cards that prompted them to provide their own reason for voting by completing the sentence, "I will vote because ______," whereas other individuals completed pledge cards that did not contain this sentence prompt. We conducted a large scale survey of individuals who filled out pledge cards and determined that receiving a pledge card with the sentence prompt had a positive influence on voting. Moreover, this effect was found above and beyond demographic and psychological predictors of voting. Implications of these findings are discussed. PMID- 10810075 TI - Poverty as we know it: media portrayals of the poor. PMID- 10810076 TI - Context effects in national health surveys: effects of preceding questions on reporting serious difficulty seeing and legal blindness. PMID- 10810078 TI - The polls-trends: terrorism in the United States. PMID- 10810077 TI - Telescoping of landmark events: implications for survey research. PMID- 10810079 TI - Comment. PMID- 10810080 TI - Reply to singer. PMID- 10810081 TI - Drosophila melanogaster: a genome sequenced. PMID- 10810082 TI - Structure of chromosomal duplicons and their role in mediating human genomic disorders. AB - Chromosome-specific low-copy repeats, or duplicons, occur in multiple regions of the human genome. Homologous recombination between different duplicon copies leads to chromosomal rearrangements, such as deletions, duplications, inversions, and inverted duplications, depending on the orientation of the recombining duplicons. When such rearrangements cause dosage imbalance of a developmentally important gene(s), genetic diseases now termed genomic disorders result, at a frequency of 0.7-1/1000 births. Duplicons can have simple or very complex structures, with variation in copy number from 2 to >10 repeats, and each varying in size from a few kilobases in length to hundreds of kilobases. Analysis of the different duplicons involved in human genomic disorders identifies features that may predispose to recombination, including large size and high sequence identity between the recombining copies, putative recombination promoting features, and the presence of multiple genes/pseudogenes that may include genes expressed in germ cells. Most of the chromosome rearrangements involve duplicons near pericentromeric regions, which may relate to the propensity of such regions to accumulate duplicons. Detailed analyses of the structure, polymorphic variation, and mechanisms of recombination in genomic disorders, as well as the evolutionary origin of various duplicons will further our understanding of the structure, function, and fluidity of the human genome. PMID- 10810084 TI - Comparative genome mapping in the sequence-based era: early experience with human chromosome 7. AB - The success of the ongoing Human Genome Project has resulted in accelerated plans for completing the human genome sequence and the earlier-than-anticipated initiation of efforts to sequence the mouse genome. As a complement to these efforts, we are utilizing the available human sequence to refine human-mouse comparative maps and to assemble sequence-ready mouse physical maps. Here we describe how the first glimpses of genomic sequence from human chromosome 7 are directly facilitating these activities. Specifically, we are actively enhancing the available human-mouse comparative map by analyzing human chromosome 7 sequence for the presence of orthologs of mapped mouse genes. Such orthologs can then be precisely positioned relative to mapped human STSs and other genes. The chromosome 7 sequence generated to date has allowed us to more than double the number of genes that can be placed on the comparative map. The latter effort reveals that human chromosome 7 is represented by at least 20 orthologous segments of DNA in the mouse genome. A second component of our program involves systematically analyzing the evolving human chromosome 7 sequence for the presence of matching mouse genes and expressed-sequence tags (ESTs). Mouse specific hybridization probes are designed from such sequences and used to screen a mouse bacterial artificial chromosome (BAC) library, with the resulting data used to assemble BAC contigs based on probe-content data. Nascent contigs are then expanded using probes derived from newly generated BAC-end sequences. This approach produces BAC-based sequence-ready maps that are known to contain a gene(s) and are homologous to segments of the human genome for which sequence is already available. Our ongoing efforts have thus far resulted in the isolation and mapping of >3,800 mouse BACs, which have been assembled into >100 contigs. These contigs include >250 genes and represent approximately 40% of the mouse genome that is homologous to human chromosome 7. Together, these approaches illustrate how the availability of genomic sequence directly facilitates studies in comparative genomics and genome evolution. PMID- 10810085 TI - Alu elements support independent origin of prosimian, platyrrhine, and catarrhine Mhc-DRB genes. AB - The primate major histocompatibility complex (Mhc) genes fall into two classes and each of the classes into several families. Of the class II families, the DRB family has a long and complex evolutionary history marked by gene turnover, rearrangement, and molecular convergence. Because the history is not easily decipherable from sequences alone, Alu element insertions were used as cladistic markers to support the surmised phylogenetic relationships among the DRB genes. Intron 1 segments of 24 DRB genes from five platyrrhine species and five DRB genes from three prosimian species were amplified by PCR and cloned, and the amplification products were sequenced or PCR-typed for Alu repeats. Three Alu elements were identified in the platyrrhine and four in the prosimian DRB genes. One of the platyrrhine elements (Alu50J) is also found in the Catarrhini, whereas the other two (Alu62Sc, Alu63Sc) are restricted to the New World monkeys. Similarly, the four prosimian elements are found only in this taxon. This distribution of Alu elements is consistent with the phylogeny of the DRB genes as determined from their intron 1 sequences in an earlier and the present study. It contradicts the exon 2-based phylogeny and thus corroborates the conclusion that the evolution of DRB exon 2 sequences is, to some extent, shaped by molecular convergence. Taken together, the data indicate that each of the assemblages of DRB genes in prosimians, platyrrhines, and catarrhines is derived from a separate ancestral gene. PMID- 10810083 TI - MHC class II pseudogene and genomic signature of a 32-kb cosmid in the house finch (Carpodacus mexicanus). AB - Large-scale sequencing studies in vertebrates have thus far focused primarily on the genomes of a few model organisms. Birds are of interest to genomics because of their much smaller and highly streamlined genomes compared to mammals. However, large-scale genetic work has been confined almost exclusively to the chicken; we know little about general aspects of genomes in nongame birds. This study examines the organization of a genomic region containing an Mhc class II B gene in a representative of another important lineage of the avian tree, the songbirds (Passeriformes). We used a shotgun sequencing approach to determine the sequence of a 32-kb cosmid insert containing a strongly hybridizing Mhc fragment from house finches (Carpodacus mexicanus). There were a total of three genes found on the cosmid clone, about the gene density expected for the mammalian Mhc: a class II Mhc beta-chain gene (Came-DAB1), a serine-threonine kinase, and a zinc finger motif. Frameshift mutations in both the second and third exons of Came DAB1 and the unalignability of the gene after the third exon suggest that it is a nonfunctional pseudogene. In addition, the identifiable introns of Came-DAB1 are more than twice as large as those of chickens. Nucleotide diversity in the peptide-binding region of Came-DAB1 (Pi = 0.03) was much lower than polymorphic chicken and other functional Mhc genes but higher than the expected diversity for a neutral locus in birds, perhaps because of hitchhiking on a selected Mhc locus close by. The serine-threonine kinase gene is likely functional, whereas the zinc finger motif is likely nonfunctional. A paucity of long simple-sequence repeats and retroelements is consistent with emerging rules of chicken genomics, and a pictorial analysis of the "genomic signature" of this sequence, the first of its kind for birds, bears strong similarity to mammalian signatures, suggesting common higher-order structures in these homeothermic genomes. The house finch sequence is among a very few of its kind from nonmodel vertebrates and provides insight into the evolution of the avian Mhc and of avian genomes generally. PMID- 10810086 TI - Phylogenetic origin of human chromosomes 7, 16, and 19 and their homologs in placental mammals. AB - The origin of human chromosomes (HSA) 7, 16, and 19 was studied by comparing data obtained from chromosome banding, chromosome painting, and gene mapping in species belonging to 11 orders of placental mammals (Eutherians). This allowed us to propose the reconstruction of their presumed ancestral forms. The HSA7 homologs were composed of two parts, the largest forming an acrocentric. The smallest formed one arm of a small submetacentric; the other arm was composed of sequences homologous to the short arm of HSA16 (HSA16p). The sequences homologous to the long arm of HSA16 (HSA16q) were associated with sequences homologous to the long arm of HSA19 (HSA19q) and formed another submetacentric. From their origin, these chromosomes underwent the following rearrangements to give rise to current human chromosomes: centromeric fission of the two submetacentrics in ancestors of all primates (approximately 80 million years ago); fusion of the HSA19p and HSA19q sequences, originating the current HSA19, in ancestors of all simians (approximately 55 million years ago); fusions of the HSA16p and HSA16q sequences, originating the current HSA16 and the two components of HSA7 before the separation of Cercopithecoids and Hominoids ( approximately 35 million years ago); and finally, pericentric and paracentric inversions of the homologs to HSA7 after the divergence of orangutan and gorilla, respectively. Thus, compared with HSA16 and HSA19, HSA7 is a fairly recent chromosome shared by man and chimpanzee only. PMID- 10810087 TI - Association between nuclear lamin A/C R482Q mutation and partial lipodystrophy with hyperinsulinemia, dyslipidemia, hypertension, and diabetes. AB - Nuclear lamins A and C are encoded by LMNA and are present in terminally differentiated cells. Lamins participate in DNA replication, chromatin organization, arrangement of nuclear pores, nuclear growth, and anchorage of nuclear membranes. In several Canadian probands with partial lipodystrophy, since found to have a common ancestor, we identified a rare novel LMNA mutation, R482Q, that completely cosegregated with the partial lipodystrophy phenotype. We evaluated the relationship between quantitative metabolic phenotypes in both diabetic and nondiabetic carriers of LMNA R482Q and family controls, who were LMNA R482/R482 homozygotes. We found that when compared with LMNA R482/R482 homozygotes: (1) diabetic LMNA Q482/R482 heterozygotes had significantly higher glucose, glycosylated hemoglobin, triglycerides, insulin and C-peptide, and significantly lower HDL cholesterol; and (2) nondiabetic LMNA Q482/R482 heterozygotes had significantly higher triglycerides, insulin and C-peptide, and significantly lower HDL cholesterol. We also found that diabetic LMNA Q482/R482 heterozygotes were older and more likely to take antihypertensive medications. Thus, LMNA R482Q was associated with lipodystrophy, hyperinsulinemia, dyslipidemia, diabetes, and hypertension. The results indicate that perturbations in plasma lipids precede the plasma glucose abnormalities in LMNA Q482-associated hyperinsulinemia. Thus, rare mutations in a nuclear structural protein can be associated with markedly abnormal qualitative and quantitative metabolic phenotypes PMID- 10810088 TI - Null mutation of PCLN-1/Claudin-16 results in bovine chronic interstitial nephritis. AB - Inherited chronic renal diseases are associated with failures in glomerular filtration and tubular resorption. Such failures invariably result from defects in selective filtration and absorption in surface renal epithelium. Recently, we described an autosomal recessive chronic interstitial nephritis with diffuse zonal fibrosis (CINF) in cattle. Bovine CINF, characterized by increased blood urea nitrogen, creatinine, and urinary proteins, leads to lethality before puberty, usually within the first 6 months or year of life. Here, we demonstrate that the first four exons of PCLN-1/Claudin-16 (CL-16), which encodes a member of the claudin family of tight junction proteins, were deleted in CINF-affected cattle. CL-16 was expressed preferentially in kidneys of normal cattle, but transcripts were totally absent in affected offspring. This observation suggests that the lack of CL-16 protein contributes to the dysfunction of paracellular renal transport systems. PMID- 10810089 TI - Comparative genomic sequencing identifies novel tissue-specific enhancers and sequence elements for methylation-sensitive factors implicated in Igf2/H19 imprinting. AB - A differentially methylated region (DMR) and endoderm-specific enhancers, located upstream and downstream of the mouse H19 gene, respectively, are known to be essential for the reciprocal imprinting of Igf2 and H19. To explain the same imprinting patterns in non-endodermal tissues, additional enhancers have been hypothesized. We determined and compared the sequences of human and mouse H19 over 40 kb and identified 10 evolutionarily conserved downstream segments, 2 of which were coincident with the known enhancers. Reporter assays in transgenic mice showed that 5 of the other 8 segments functioned as enhancers in specific mesodermal and/or ectodermal tissues. We also identified a conserved 39-bp element that appeared repeatedly within the DMR and formed complexes with specific nuclear factors. Binding of one of the factors was inhibited when the target sequence contained methylated CpGs. These complexes may contribute to the presumed boundary function of the unmethylated DMR, which is proposed to insulate maternal Igf2 from the enhancers. Our results demonstrate that comparative genomic sequencing is highly efficient in identifying regulatory elements. PMID- 10810090 TI - Nature and structure of human genes that generate retropseudogenes. AB - The human genome is estimated to contain 23,000 to 33,000 retropseudogenes. To study the properties of genes giving rise to these retroelements, we compared the structure and expression of genes with or without known retropseudogenes. Four main features have emerged from the analysis of 181 genes associated to retropseudogenes: Reverse-transcribed genes are (1) widely expressed, (2) highly conserved, (3) short, and (4) GC-poor. The first two properties probably reflect the fact that genes giving rise to retropseudogenes have to be expressed in the germ-line. The two latter points suggest that reverse-transcription and transposition is more efficient for short GC-poor mRNAs. In addition, this analysis allowed us to reject previous hypotheses that widely expressed genes are GC rich. Rather, globally, genes with a wide tissue distribution are GC poor. PMID- 10810091 TI - Analysis of 5S rDNA arrays in Arabidopsis thaliana: physical mapping and chromosome-specific polymorphisms. AB - A physical map of a pericentromeric region of chromosome 5 containing a 5S rDNA locus and spanning approximately 1000 kb was established using the CIC YAC clones. Three 5S rDNA arrays were resolved in this YAC contig by PFGE analysis and we have mapped different types of sequences between these three blocks. 5S rDNA units from each of these three arrays of chromosome 5, and from chromosomes 3 and 4, were isolated by PCR. A total of 38 new DNA sequences were obtained. Two types of 5S rDNA repeated units exist: the major variant with 0.5-kb repeats and one with short repeats (251 bp) only detected on YAC 11A3 from chromosome 3. Although the 38 sequences displayed noticeable heterogeneity, we were able to group them according to their 5S array origin. The presence of 5S array-specific variants was confirmed with the restriction polymorphism study of all the YACs carrying 5S units. PMID- 10810092 TI - A radiation hybrid map of the cat genome: implications for comparative mapping. AB - Ordered gene maps of mammalian species are becoming increasingly valued in assigning gene variants to function in human and animal models, as well as recapitulating the natural history of genome organization. To extend this power to the domestic cat, a radiation hybrid (RH) map of the cat was constructed integrating 424 Type I-coding genes with 176 microsatellite markers, providing coverage over all 20 feline chromosomes. Alignment of parallel RH maps of human and cat reveal 100 conserved segments ordered (CSOs) between the species, nearly three times the number observed with reciprocal chromosome painting analyses. The observed number is equivalent to theoretical predictions of the number of conserved segments to be found between cat and human, implying that 300-400 Type I gene markers is sufficient to reveal nearly all conserved segments for species that exhibit the most frequently observed "slow" rate of genome reorganization. The cat-human RH map comparisons provide a new genomic tool for comparative gene mapping in the cat and related Felidae, and provide confirmation that the cat genome organization is remarkably conserved compared with human. These data demonstrate that ordered RH-based gene maps provide the most precise assessment of comparing genomes, short of contig construction or full-sequence determination. PMID- 10810093 TI - Identification of novel human genes evolutionarily conserved in Caenorhabditis elegans by comparative proteomics. AB - Modern biomedical research greatly benefits from large-scale genome-sequencing projects ranging from studies of viruses, bacteria, and yeast to multicellular organisms, like Caenorhabditis elegans. Comparative genomic studies offer a vast array of prospects for identification and functional annotation of human ortholog genes. We presented a novel comparative proteomic approach for assembling human gene contigs and assisting gene discovery. The C. elegans proteome was used as an alignment template to assist in novel human gene identification from human EST nucleotide databases. Among the available 18,452 C. elegans protein sequences, our results indicate that at least 83% (15,344 sequences) of C. elegans proteome has human homologous genes, with 7,954 records of C. elegans proteins matching known human gene transcripts. Only 11% or less of C. elegans proteome contains nematode-specific genes. We found that the remaining 7,390 sequences might lead to discoveries of novel human genes, and over 150 putative full-length human gene transcripts were assembled upon further database analyses. [The sequence data described in this paper have been submitted to the PMID- 10810094 TI - Construction of a BAC contig map of chromosome 16q by two-dimensional overgo hybridization. AB - We have used sequence-based markers from an integrated YAC STS-content/somatic cell hybrid breakpoint physical map and radiation hybrid maps of human chromosome 16 to construct a new sequence-ready BAC map of the long arm of this chromosome. The integrated physical map was generated previously in our laboratory and contains 1150 STSs, providing a marker on average every 78 kb on the euchromatic arms of chromosome 16. The other two maps used for this effort were the radiation hybrid maps of chromosome 16 from Whitehead Institute and Stanford University. To create large sequenceable targets of this chromosome, we used a systematic approach to screen high-density BAC filters with probes generated from overlapping oligonucleotides (overgos). We first identified all available sequences in the three maps. These include sequences from genes, ESTs, STSs, and cosmid end sequences. We then used BLASTto identify 36-bp unique fragments of DNA for overgo probes. A total of 906 overgos were selected from the long arm of chromosome 16. Hybridizations occurred in three stages: (1) superpool hybridizations against the 12x coverage human BAC library (RPCI-11); (2) two dimensional hybridizations against rearrayed positive BACs identified in the superpool hybridizations; and (3) pooled tertiary hybridizations for those overgos that had ambiguous positives remaining after the two-dimensional hybridization. For the superpool hybridizations, up to 236 overgos have been pooled in a single hybridization against the 12x BAC library. A total of 5187 positive BACs from chromosome 16q were identified as a result of five superpool hybridizations. These positive clones were rearrayed on membranes and hybridized with 161 two-dimensional subpools of overgos to determine which BAC clones were positive for individual overgos. An additional 46 tertiary hybridizations were required to resolve ambiguous overgo-BAC relationships. Thus, after a total of 212 hybridizations, we have constructed an initial probe-content BAC map of chromosome 16q consisting of 828 overgo markers and 3363 BACs providing >85% coverage of the long arm of this chromosome. The map has been confirmed by the fingerprinting data and BAC end PCR screening. PMID- 10810095 TI - Living near opencast coal mining sites and children's respiratory health. AB - OBJECTIVES: To answer the question whether living near opencast coal mining sites affects acute and chronic respiratory health. METHODS: All 4860 children aged 1 11 from five socioeconomically matched pairs of communities close to active opencast sites and control sites away from them were selected. Exposure was assessed by concentrations of particulate matter with aerodynamic diameter < 10 microns (PM10), residential proximity to active opencast sites, and particle composition. PM10 was monitored and sampled for 6 weeks in four pairs, and for 24 weeks in one pair. A postal questionnaire collected data on health and lifestyle. Daily health information was collected by a symptom diary (concurrently with PM10 monitoring) and general practitioner (GP) records were abstracted (concurrently with PM10 monitoring and 52 weeks before the study). Outcomes were the cumulative and period prevalence (2 and 12 months) of wheeze, asthma, bronchitis, and other respiratory symptoms, and the prevalence and incidence of daily symptoms and GP consultations. RESULTS: Patterns of the daily variation of PM10 were similar in opencast and control communities, but PM10 was higher in opencast areas (mean ratio 1.14, 95% confidence interval (95% CI) 1.13 to 1.16, geometric mean 17.0 micrograms/m3 v 14.9 micrograms/m3). Opencast sites were a measurable contributor to PM10 in adjacent areas. Little evidence was found for associations between living near an opencast site and an increased prevalence of respiratory illnesses, asthma severity, or daily diary symptoms, but children in opencast communities 1-4 had significantly more respiratory consultations (1.5 v 1.1 per person-year) than children in control communities for the 6 week study periods. Associations between daily PM10 concentrations and acute health events were similar in opencast and control communities. CONCLUSIONS: Children in opencast communities were exposed to a small but significant amount of additional PM10 to which the opencast sites were a measurable contributor. Past and present respiratory health of children was similar, but GP consultations for respiratory conditions were higher in opencast communities during the core study period. PMID- 10810096 TI - Local road traffic activity and the prevalence, severity, and persistence of wheeze in school children: combined cross sectional and longitudinal study. AB - OBJECTIVES: To investigate the relation between local road traffic activity and the occurrence, severity, and persistence of wheeze in children. METHODS: Data on wheeze and asthma were collected in a cross sectional questionnaire survey of 22,968 primary school children (age 4-11) and 27,826 secondary school children (age 11-16) in the Nottingham area. Direct measures of road traffic flow were made in the locality of each school and combined with Local Authority traffic data for major roads to estimate local traffic activity in vehicle metres/day/km2. Assessment of the effects of potential confounders was performed in nested case-control groups of 6576 primary and 5936 secondary children. Data on frequency of wheeze were collected for the cases to study disease severity. Longitudinal data on a historical cohort of 883 children who reported wheeze when aged 4-11 in 1988 were used to study the persistence of wheeze into adolescence. RESULTS: Unadjusted prevalence of wheeze in the past year within schools varied widely but was not associated with traffic activity in the school locality (weighted regression coefficient beta = -0.01, p = 0.93 for primary schools, beta = -0.18, p = 0.26 for secondary schools). The risk of wheeze in individual primary school children was not associated with traffic activity analysed as a continuous variable, although there was some suggestion of a weak, non-linear plateau effect. Similar effects were found for diagnosed asthma and recent cough. There was no evidence of any relation between traffic activity and risk of wheeze in secondary school children. There were positive but non-significant dose related effects of traffic activity on wheeze severity in primary and secondary children and on persistence of wheeze in the longitudinal cohort. CONCLUSIONS: Traffic activity in the school locality is not a major determinant of wheeze in children. PMID- 10810097 TI - Detection of specific antibodies to pigeon serum and bloom antigens by enzyme linked immunosorbent assay in pigeon breeder's disease. AB - BACKGROUND: Pigeon breeder's disease is an extrinsic allergic alveolitis in the lungs of sensitised people, caused by hypersensitivity reactions to inhaled pigeon antigens. Antigens from different sources of the animal are used for diagnostic purposes, with serum being the most widely used. Bloom is rarely used; very little is known of its antigenicity and diagnostic performance, particularly when used with the enzyme linked immunosorbent assay (ELISA) method, which is the most popular test as it permits measurement of the antibody response. METHODS: To (a) standardise an ELISA for the measurement of specific IgG against pigeon serum and pigeon bloom extract; (b) to establish reference values for specific IgG in 73 non-exposed controls, (c) to show the presence of specific IgG against pigeon serum and bloom in serum samples of 17 patients with bird fancier's lung and 11 asymptomatic fanciers, and (d) to study the similarity of the two antigen sources by cross reactivity experiments. RESULTS: Reference values of specific IgG were defined with the 97.5 percentile (367.9 U/ml for pigeon serum and 953.7 U/ml for pigeon bloom extract). Of symptomatic patients 100% had values higher than the cut off for both antigens. In asymptomatic fanciers values were higher than the cut off for pigeon serum in 45% and bloom extract in 54%. Cross reactivity experiments showed that the two antigens differed in antigenic content although some components may be common to both. CONCLUSION: The ELISA methods used proved to be useful tools for evaluating specific IgG antibody responses against both antigens. The diagnostic performance of both ELISA methods performed with these antigen sources was similar, showing very high sensitivity but moderate specificity. Although some antigenic similarity was found between pigeon serum and bloom extract, cross reactivity studies showed that various antigens seemed to be specific to the bloom extract. However, the antigens responsible for pigeon breeder's disease seem to be present in both antigenic sources. PMID- 10810098 TI - Heat exhaustion in a deep underground metalliferous mine. AB - OBJECTIVES: To examine the incidence, clinical state, personal risk factors, haematology, and biochemistry of heat exhaustion occurring at a deep underground metalliferous mine. To describe the underground thermal conditions associated with the occurrence of heat exhaustion. METHODS: A 1 year prospective case series of acute heat exhaustion was undertaken. A history was obtained with a structured questionnaire. Pulse rate, blood pressure, tympanic temperature, and specific gravity of urine were measured before treatment. Venous blood was analysed for haematological and biochemical variables, during the acute presentation and after recovery. Body mass index (BMI) and maximum O2 consumption (VO2 max) were measured after recovery. Psychrometric wet bulb temperature, dry bulb temperature, and air velocity were measured at the underground sites where heat exhaustion had occurred. Air cooling power and psychrometric wet bulb globe temperature were derived from these data. RESULTS: 106 Cases were studied. The incidence of heat exhaustion during the year was 43.0 cases/million man-hours. In February it was 147 cases/million man-hours. The incidence rate ratio for mines operating below 1200 m compared with those operating above 1200 m was 3.17. Mean estimated fluid intake was 0.64 l/h (SD 0.29, range 0.08-1.50). The following data were increased in acute presentation compared with recovery (p value, % of acute cases above the normal clinical range): neutrophils (p < 0.001, 36%), anion gap (p < 0.001, 63%), urea (p < 0.001, 21%), creatinine (p < 0.001, 30%), glucose (p < 0.001, 15%), serum osmolality (p = 0.030, 71%), creatine kinase (p = 0.002, 45%), aspartate transaminase (p < 0.001, 14%), lactate dehydrogenase (p < 0.001, 9.5%), and ferritin (p < 0.001, 26%). The following data were depressed in acute presentation compared with recovery (p value, % of acute cases below the normal clinical range): eosinophils (p = 0.003, 38%) and bicarbonate (p = 0.011, 32%). Urea and creatinine were significantly increased in miners with heat cramps compared with miners without this symptom (p < 0.001), but there was no significant difference in sodium concentration (p = 0.384). Mean psychrometric wet bulb temperature was 29.0 degrees C (SD 2.2, range 21.0-34.0). Mean dry bulb temperature was 37.4 degrees C (SD 2.4, range 31.0-43.0). Mean air velocity was 0.54 m/s (SD 0.57, range 0.00-4.00). Mean air cooling power was 148 W/m2 (SD 49, range 33-290) Mean psychrometric wet bulb globe temperature was 31.5 degrees C (SD 2.0, range 25.2-35.3). Few cases (< 5%) occurred at psychrometric wet bulb temperature < 25.0 degrees C, dry bulb temperature < 33.8 degrees C, air velocity > 1.56 m/s, air cooling power > 248 W/m2, or psychrometric wet bulb globe temperature < 28.5 degrees C. CONCLUSION: Heat exhaustion in underground miners is associated with dehydration, neutrophil leukocytosis, eosinopenia, metabolic acidosis, increased glucose and ferritin, and a mild rise in creatine kinase, aspartate transaminase, and lactate dehydrogenase. Heat cramps are associated with dehydration but not hyponatraemia. The incidence of heat exhaustion increases during summer and at depth. An increased fluid intake is required. Heat exhaustion would be unlikely to occur if ventilation and refrigeration achieved air cooling power > 250 W/m2 at all underground work sites. PMID- 10810099 TI - Incidence of cancer among commercial airline pilots. AB - OBJECTIVES: To describe the cancer pattern in a cohort of commercial pilots by follow up through the Icelandic Cancer Registry. METHODS: This is a retrospective cohort study of 458 pilots with emphasis on subcohort working for an airline operating on international routes. A computerised file of the cohort was record linked to the Cancer Registry by making use of personal identification numbers. Expected numbers of cancer cases were calculated on the basis of number of person years and incidences of cancer at specific sites for men provided by the Cancer Registry. Numbers of separate analyses were made according to different exposure variables. RESULTS: The standardised incidence ratio (SIR) for all cancers was 0.97 (95% confidence interval (95% CI) 0.62 to 1.46) in the total cohort and 1.16 (95% CI 0.70 to 1.81) among those operating on international routes. The SIR for malignant melanoma of the skin was 10.20, 95% CI 3.29 to 23.81 in the total cohort and 15.63, 95% CI 5.04 to 36.46 in the restricted cohort. Analyses according to number of block-hours and radiation dose showed that malignant melanomas were found in the subgroups with highest exposure estimates, the SIRs were 13.04 and 28.57 respectively. The SIR was 25.00 for malignant melanoma among those who had been flying over five time zones. CONCLUSIONS: The study shows a high occurrence of malignant melanoma among pilots. It is open to discussion what role exposure of cosmic radiation, numbers of block-hours flown, or lifestyle factors--such as possible excessive sunbathing--play in the aetiology of cancer among pilots. This calls for further and more powerful studies. The excess of malignant melanoma among those flying over five time zones suggests that the importance of disturbance of the circadian rhythm should be taken into consideration in future studies. PMID- 10810100 TI - Exposure to high concentrations of nitrosamines and cancer mortality among a cohort of rubber workers. AB - OBJECTIVES: To examine if the occurrence of different cancers was increased among rubber workers, as the highest known exposures of humans to nitrosamines have occurred in the rubber industry. METHODS: A cohort of 8933 rubber workers (hired after 1 January 1950, still active or retired on 1 January 1981 and employed for at least 1 year in one of five study factories) was followed up for mortality from 1 January 1981 to 31 December 1991. Work histories were reconstructed with routinely documented cost centre codes, which allowed identification by employment in specific work areas. For each cost centre code time and factory specific, semi-quantitative exposures to nitrosamines (three levels: low, medium, high) and other compounds were estimated by industrial hygienists. Rate ratios for medium (RRm) and high (RRh) exposures and 95% confidence intervals (95% CIs) were calculated with Cox's proportional hazards models with the low exposure as reference. RESULTS: Exposure to nitrosamines was significantly associated with an increased mortality from cancers of the oesophagus (13 deaths: RRm 1.7, 95% CI 0.3 to 10.3; RRh 7.3, 95% CI 1.9 to 27.8) and of the oral cavity and pharynx (17 deaths: RRm 0.8, 95% CI 0.2 to 4.1; RRh 3.9, 95% CI 1.4 to 11.1). A non significant trend of increasing mortality with exposure to higher concentrations of nitrosamines was found for mortality from cancer of the prostate (26 deaths: RRm 1.4, 95% CI 0.5 to 3.8; RRh 2.2, 95% CI 0.9 to 5.6), and the brain (six deaths: RRm 3.9, 95% CI 0.3 to 42.6; RRh 6.0, 95% CI 0.6 to 57.6). No association was found between exposure to nitrosamines and cancer of the stomach (RRm 0.8, 95% CI 0.4 to 1.8; RRh 1.2, 95% CI 0.5 to 2.5) or lung (RRm 1.0, 95% CI 0.6 to 1.5; RRh 1.0, 95% CI 0.7 to 1.6). CONCLUSIONS: Exposure to high concentrations of nitrosamines is associated with increased mortality from cancers of the oesophagus, oral cavity, and pharynx, but not with increased mortality from cancers of the stomach or lung. PMID- 10810101 TI - Epidemiological response to a suspected excess of cancer among a group of workers exposed to multiple radiological and chemical hazards. AB - OBJECTIVE: An excess of cancer was suspected by workers of the metallurgy department at the French Atomic Energy Commissariat (CEA) after several deaths from cancer were reported in 1983 and 1984. After a descriptive study performed by the CEA in 1985 the results of which were not conclusive enough to put an end to the controversy, the present cohort study was undertaken in 1989. METHODS: As no specific exposure, or a precise cancer site was suspected, it was decided to include all subjects who had worked at the metallurgy department for at least 1 year between 1950 and 1968. The cohort was followed up to 31 December 1990. Individual occupational exposures were determined retrospectively for each year from 1950 to 1990, both qualitatively (annual job, and hazard records, and assistance from former workers) and quantitatively (for external radiation). On the basis of these exposures, three types of occupational tasks were identified: handling of chemicals, radionuclides, and external radiation. Standardised mortality ratios (SMRs) were calculated to estimate the risk of death, and the existence of an association between risk of cancer and each of the three tasks was tested. RESULTS: The cohort included 356 workers, followed up for an average of 30 years (total of 10,820 person-years). The number of deaths from all causes and from all cancer sites were respectively 44 and 21. No excess of cancer deaths was found for the study period (SMR 0.77), nor was there a peak in 1983-4. The risk of death from all cancer sites increased with the duration of exposure to chemicals. CONCLUSION: The results do not justify the workers' impression of an excess of cancer. They suggest, however, that the duration of work at some tasks that involved handling chemicals may be an indicator of risk of cancer. Communication to the workers during the study played an important part in reducing their concern, contributing to their better understanding of the results. PMID- 10810102 TI - Self reported symptoms and inhibition of acetylcholinesterase activity among Kenyan agricultural workers. AB - OBJECTIVES: This study was part of the East African pesticides project. The general objective was to assess health hazards posed by handling, storage, and use of pesticides, on agricultural estates and small farms with a view to developing strategies for prevention and control of pesticide poisoning. The aim of this paper is to describe the prevalence of symptoms in this population, to relate levels of inhibition to reported symptoms and evaluate at which levels of inhibition symptoms become increased. METHODS: Complete data were available for 256 exposed subjects and 152 controls from four regions in Kenya. A structured questionnaire on symptoms experienced at the time of interview was given to all subjects and controls. Information was also obtained on sex, age, main occupation, and level of education. Symptoms reported during the high exposure period, were initially clustered in broader symptom categories from reference literature on health effects of pesticides that inhibit cholinesterase (organophosphate and carbamate). Prevalence ratios were estimated for symptoms with changes in cholinesterase activity in serum. RESULTS: Symptom prevalence in exposed subjects was higher during the high exposure period than the low exposure period, although these differences were not significant. Interestingly, a clear and significant change in symptoms prevalence was found in the controls with a higher prevalence in the low exposure period. Analysis of the relation between cholinesterase inhibition and symptoms showed that prevalence ratios were significantly > 1 for respiratory, eye, and central nervous system symptoms for workers with > 30% inhibition. Similar results were found for analyses with the actual level of acetylcholinesterase activity. CONCLUSION: The results suggest the presence of a relation between exposure and acetylcholinesterase inhibition, acetylcholinesterase activity, and respiratory, eye, and central nervous system symptoms. Increased symptom prevalence was found at acetylcholinesterase activities generally considered to be non-adverse. PMID- 10810103 TI - Effect of exposure to lead on postural control in workers. AB - OBJECTIVES: To examine the effect of lead on postural control of workers who have been exposed to lead. METHODS: 63 Male, lead battery workers mean (SD) age 41.0 (7.4) were compared with 48 age matched male controls after excluding those with acute or chronic diseases. Exposed workers had mean (SD) past blood lead concentrations of 37.5 (9.2) micrograms/dl and 11.2 (5.7) years of employment. Postural control was measured with a computerised postural sway measurement system which measured both sway and total movements. RESULTS: Workers standing straight with eyes open on the bare plates had sway and total movements which were not notably different from controls. On the other hand increased movements were needed in the exposed workers to maintain stability (the general stability quotient 18.2 (5.4) v 15.4 (4.4) in controls, p < 0.01) when standing directly on the foot-plates with closed eyes,, and with the head tilted (15.0 (3.8) v 11.5 (3.0) in controls, p < 0.001). Exposed workers also had a trend for less ability to synchronize anterior posterior and lateral sway in the stress positions (0.0625) than had non-exposed workers. Significant but low correlations were found between the estimate of the chronic internal dose of lead and three of 10 of the postural control measurements, and present lead blood concentrations and only one of the 10 measurements and (r values ranged from 0.21 to 0.31, p < or = 0.03). CONCLUSIONS: These findings suggest that lead affects postural control in asymptomatic workers. Further studies are warranted to find whether workers with decreased postural control are at increased risk of accidents and the relation, if any, of these measurements with subsequent morbidity. PMID- 10810104 TI - Polychlorinated dibenzo-p-dioxin and dibenzofuran concentrations in the serum samples of workers at continuously burning municipal waste incinerators in Japan. AB - OBJECTIVES: To find whether concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in serum increased in workers at municipal incinerators that burn continuously. METHODS: 30 Workers employed at three municipal waste incineration plants (incinerator workers) and 30 control workers were studied. The incinerator workers had worn dust masks or airline masks during the periodic repair work inside the incinerators. Previous job, dietary habit, smoking habit, distance from residence to the incineration plant, and body weight and height were obtained from a questionnaire survey. Concentrations of PCDDs/PCDFs were measured in the serum of the workers and the dust deposited in the plants. The influence of various factors on serum concentrations of PCDDs/PCDFs was examined by multiple regression analysis. RESULTS: Dust analysis showed the greatest amount of octachlorodibenzo-p-dioxin (OCDD), followed by 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD), 1,2,3,4,6,7,8-heptachlorodibenzofuran (HpCDF), and octachlorodibenzofuran (OCDF). The toxicity equivalents (TEQs) of PCDDs and PCDFs in the deposited dust were 4.8, 1.0, and 6.4 ng TEQs/g, respectively, for plants A, B, and C. The mean serum TEQs of PCDDs and PCDFs in the incinerator workers and control workers were 19.2 and 22.9 pg TEQs/g lipid, respectively, for area A, 28.8 and 24.5 pg TEQs/g lipid for area B, and 23.4 and 23.6 pg TEQs/g lipid for area C. No significant differences were found between the incinerator workers and the controls for TEQs of PCDDs and PCDFs separately, and TEQs of PCDDs and PCDFs together. However, the serum 1,2,3,4,6,7,8-HpCDF concentration was significantly higher in the incinerator workers than in the controls for all the three areas. When the exposure index to 1,2,3,4,6,7,8-HpCDF is defined as the product of the concentration of 1,2,3,4,6,7,8-HpCDF in the deposited dust and duration of employment, the concentration of 1,2,3,4,6,7,8-HpCDF in serum increased as the exposure index increased. Multivariate analysis suggested that the serum concentration of HpCDF increased with duration of employment at the incineration plants and OCDF increased with employment of > or = 21 years. The other significant variables (p < 0.01 or p < 0.001) were area for hexachlorodibenzo-p dioxin (HxCDD) and tetrachlorodibenzofuran (TCDF), Brinkman index for HpCDD, and body mass index (BMI) for tetrachlorodibenzo-p-dioxin (TCDD), HpCDD, and TEQs of PCDDs. CONCLUSION: The serum TEQs of PCDDs and PCDFs was not significantly higher among the incinerator workers, but the serum concentration of 1,2,3,4,6,7,8-HpCDF was. This suggests that the incinerator workers had inhaled dust containing PCDDs and PCDFs while working in plants equipped with incinerators that burn continuously. PMID- 10810105 TI - Associations of self estimated workloads with musculoskeletal symptoms among hospital nurses. AB - OBJECTIVES: To investigate the prevalence of neck, shoulder, and arm pain (NSAP) as well as low back pain (LBP) among hospital nurses, and to examine the association of work tasks and self estimated risk factors with NSAP and LBP. METHODS: A cross sectional study was carried out in a national university hospital in Japan. Full time registered nurses in the wards (n = 314) were selected for analysis. The questionnaire was composed of items on demographic conditions, severity of workloads in actual tasks, self estimated risk factors for fatigue, and musculoskeletal pain in the previous month. Rate ratios (RRs) and 95% confidence intervals (95% CIs) were calculated by the Cox's proportional hazards model to study the association of pain with variables related to work and demographic conditions. RESULTS: The prevalences of low back, shoulder, neck, and arm pain in the previous month were 54.7%, 42.8%, 31.3%, and 18.6%, respectively. The prevalence of musculoskeletal symptoms among hospital nurses was higher than in previous studies. In the Cox's models for LBP and NSAP, there were no significant associations between musculoskeletal pain and the items related to work and demographic conditions. The RRs for LBP tended to be relatively higher for "accepting emergency patients" and some actual tasks. Some items of self estimated risk factors for fatigue tended to have relatively higher RRs for LBP and NSAP. CONCLUSIONS: It was suggested that musculoskeletal pain among hospital nurses may have associations with some actual tasks and items related to work postures, work control, and work organisation. Further studies, however, are necessary, as clear evidence of this potential association was not shown in the study. PMID- 10810106 TI - Occupational and environmental medicine: on line and further developments to come: www.oem.bmjjournals.com. PMID- 10810107 TI - Prevalence and pattern of occupational exposure to hand transmitted vibration in Great Britain: findings from a national survey. AB - OBJECTIVES: To estimate the number of workers in Great Britain with significant occupational exposure to hand transmitted vibration (HTV). Also, to identify the occupations and industries where such exposures arise, and the main sources of exposure. METHODS: A questionnaire was posted to 22,194 men and women aged 16-64, comprising 21,201 subjects selected at random from the age-sex registers of 34 general practices in England, Scotland, and Wales, and a further 993 subjects selected at random from the central pay registers of the three armed services. Among other things, the questionnaire asked about exposure to sources of HTV in current and earlier employment. Responses were assessed by occupation and industry, and prevalence estimates for the country as a whole were derived from census information on occupational and industrial populations nationally. Estimates were also made in exposed workers of the average daily dose of vibration (A(8) root mean squared (rms) for the past week, based on their reported sources and durations of exposure. RESULTS: Usable questionnaires were returned by 12,907 subjects (overall response rate 58%). From these it was estimated that some 4.2 million men and 667 000 women in Great Britain are exposed to HTV at work in a 1 week period, and that personal daily exposures to vibration exceed a suggested action level equivalent to 2.8 ms(-2) for 8 hours (A(8) >2.8 ms(-2) rms) in at least 1.2 million men and 44,000 women. High estimated doses (A(8) >5 ms(-2) rms) arose most often in bricklayers and masons, gardeners and groundsmen, carpenters and joiners, electricians and electrical maintenance fitters, and builders and building contractors. The industries where high A(8) values most often arose were construction, motor vehicle repair and maintenance, manufacture of basic metals, and agriculture. The most common sources of exposure were hammer drills, hand held portable grinders, and jigsaws. CONCLUSIONS: Exposure to HTV is surprisingly prevalent, and preventive measures and health surveillance may be warranted for many men in Britain. Control strategies should focus on prevention at source, with priority accorded to the common sources of exposure and the occupations in which significant exposures tend to arise. Many vibratory tools that are common in Britain have been overlooked in previous surveys, highlighting an important focus for future research. PMID- 10810108 TI - Prevalence and pattern of occupational exposure to whole body vibration in Great Britain: findings from a national survey. AB - OBJECTIVES: To estimate the number of workers in Great Britain with significant occupational exposure to whole body vibration (WBV) and to identify the common sources of exposure and the occupations and industries where such exposures arise. METHODS: A postal questionnaire was posted to a random community sample of 22,194 men and women of working age. Among other things, the questionnaire asked about exposure to WBV in the past week, including occupational and common non occupational sources. Responses were assessed by occupation and industry, and national prevalence estimates were derived from census information. Estimates were also made of the average estimated daily personal dose of vibration (eVDV). RESULTS: From the 12,907 responses it was estimated that 7.2 million men and 1.8 million women in Great Britain are exposed to WBV at work in a 1 week period if the occupational use of cars, vans, buses, trains, and motor cycles is included within the definition of exposure. The eVDV of >374,000 men and 9000 women was estimated to exceed a proposed British Standard action level of 15 ms(-1.75). Occupations in which the estimated exposures most often exceeded 15 ms(-1.75) included forklift truck and mechanical truck drivers, farm owners and managers, farm workers, and drivers of road goods vehicles. These occupations also contributed the largest estimated numbers of workers in Great Britain with such levels of exposure. The highest estimated median occupational eVDVs were found in forklift truck drivers, drivers of road goods vehicles, bus and coach drivers, and technical and wholesale sales representatives, among whom a greater contribution to total dose was received from occupational exposures than from non occupational ones; but in many other occupations the reverse applied. The most common sources of occupational exposure to WBV are cars, vans, forklift trucks, lorries, tractors, buses, and loaders. CONCLUSIONS: Exposure to whole body vibration is common, but only a small proportion of exposures exceed the action level proposed in British standards, and in many occupations, non-occupational sources are more important than those at work. The commonest occupational sources of WBV and occupations with particularly high exposures have been identified, providing a basis for targeting future control activities. PMID- 10810109 TI - Validity of self reported occupational exposures to hand transmitted and whole body vibration. AB - OBJECTIVES: To assess the accuracy with which workers report their exposure to occupational sources of hand transmitted (HTV) and whole body vibration (WBV). METHODS: 179 Workers from various jobs involving exposure to HTV or WBV completed a self administered questionnaire about sources of occupational exposure to vibration in the past week. They were then observed at work over 1 hour, after which they completed a second questionnaire concerning their exposures during this observation period. The feasibility of reported sources of exposure during the past week was examined by questioning managers and by inspection of tools and machines in the workplace. The accuracy of reported sources and durations of exposure in the 1 hour period were assessed relative to what had been observed. RESULTS: The feasibility of exposure in the previous week was confirmed for 97% of subjects who reported exposure to HTV, and for 93% of subjects who reported exposure to WBV. The individual sources of exposure reported were generally plausible, but occupational use of cars was substantially overreported, possibly because of confusion with their use in travel to and from work. The accuracy of exposures reported during the observation period was generally high, but some sources of HTV were confused-for example, nailing and stapling guns reported as riveting hammers, and hammer drills not distinguished from other sorts of drill. Workers overestimated their duration of exposure to HTV by a median factor of 2.5 (interquartile range (IQR) 1.6-5.9), but estimated durations of exposure were more accurate when the exposure was relatively continuous rather than for intermittent short periods. Reported durations of exposure to WBV were generally accurate (median ratio of reported to observed time 1.1, IQR 1.0-1.2). CONCLUSIONS: Sources of recent occupational exposure to vibration seem to be reported with reasonable accuracy, but durations of exposure to HTV are systematically overestimated, particularly when the exposure is intermittent and for short periods. This raises the possibility that dose-response relations may have been biased in some of the studies on which exposure standards might be based, and that the levels in currently proposed standards may be too high. Future studies should pay attention to this source of error during data collection. PMID- 10810110 TI - Epidemiological survey of workers exposed to inorganic germanium compounds. AB - OBJECTIVES: To assess occupational exposure to inorganic germanium (Ge) in workers from a producing plant, and to assess the health of these workers, with a special focus on respiratory, kidney, and liver functions. METHODS: Cross sectional study of 75 workers exposed to Ge and 79 matched referents. Exposure was characterised by measuring air and urine concentrations of the element during a typical working week, and health was assessed by a questionnaire, clinical examination, lung function testing, chest radiography, and clinical chemistry in serum and urine, including high and low molecular weight urinary proteins. RESULTS: Airborne concentrations of Ge (inhalable fraction) ranged from 0.03 to 300 micrograms/m, which was reflected by increased urinary excretion of Ge (0.12 200 micrograms/g creatinine, after the shift at the end of the working week). Lung, liver, and haematological variables were not significantly different between referents and workers exposed to Ge. A slightly higher urinary concentration of high molecular weight proteins (albumin and transferrin) was found in workers exposed to Ge, possibly reflecting subclinical glomerular changes. No relation was found between the intensity or duration of exposure and the urinary concentration of albumin. No difference between referents and workers exposed to Ge was found for other renal variables. CONCLUSIONS: Measurement of urinary Ge can detect occupational exposure to inorganic Ge and its compounds. It is prudent to recommend the monitoring of renal variables in workers exposed to Ge. PMID- 10810111 TI - Non-Hodgkin's lymphoma among electric utility workers in Ontario: the evaluation of alternate indices of exposure to 60 Hz electric and magnetic fields. AB - OBJECTIVES: To examine associations between non-Hodgkin's lymphoma (NHL) and exposures to 60 Hz magnetic and electric fields in electric utility workers with a series of indices that capture a variety of aspects of field strength. METHODS: The study population consisted of 51 cases of NHL and 203 individually matched controls identified from within a cohort of male electric utility workers in Ontario. Odds ratios were calculated for several exposure indices with conditional logistic regression models. Aspects of exposure to electric and magnetic fields that were modelled included: the percentage of time spent above selected threshold field intensities, mean transitions in field strength, SD, and the arithmetic and geometric mean field intensities. RESULTS: For the most part, there was a lack of an association between exposure indices of magnetic fields and the incidence of NHL. Subjects in the upper tertile of percentage of time spent above electric field intensities of 10 and 40 V/m had odds ratios of 3.05 (95% confidence interval (95% CI) 1. 07 to 8.80) and 3.57 (1.30 to 9.80), respectively, when compared with those in the lowest tertile. Moreover, the percentages of time spent above these electric field thresholds were significant predictors of case status over and above the association explained by duration of employment and the arithmetic or geometric mean exposure. CONCLUSIONS: These data suggest that exposures above electric field threshold intensities of 10 and 40 V/m are important predictors of NHL. Consequently, the findings support the hypothesis that electric fields may play a promoting part in the aetiology of this cancer. Further occupational studies that include assessment of exposure to electric fields and measures of field strength above similar threshold cut off points are needed to confirm these findings. PMID- 10810113 TI - Cancers in concrete workers: results of a cohort study of 33,668 workers. AB - OBJECTIVES: To study cancer morbidity patterns in concrete workers. METHODS: A cohort of 33,503 concrete workers was enrolled in the study from 1971-86. The average duration of follow up was 19.4 years (582,225 person-years). The workers' cancer morbidity was compared with the morbidity of the general population. RESULTS: A total of 3572 incident cancers were observed. Significantly increased standardised incidence ratios (SIRs) were found for all malignant neoplasms (SIR 107; 95% confidence interval (95%CI) 103 to 110), cancer of the lip (SIR 179; 95%CI 134 to 234), cancer of the stomach (SIR 139; 95%CI 122 to 158), cancer of the lung (SIR 125; 95%CI 114 to 137), and cancer of the prostate (SIR 108; 95%CI 101 to 116). Reduced risk was found for cancer of the large intestine (SIR 80; 95%CI 69 to 93) and cancer of the testis (SIR 50; 95%CI 26 to 87). Smoking was more prevalent among the concrete workers than in the general population (50% v 35%). CONCLUSION: The study has shown a slightly increased overall risk of cancer among concrete workers. The increased risk of lung cancer could entirely be due to differences in smoking habits between concrete workers and the general population. There is a possibility that the smoking also has contributed to the increased risks of stomach cancer and lip cancer, but occupational factors may have contributed to these cancer sites. PMID- 10810112 TI - Exposure to electromagnetic fields and suicide among electric utility workers: a nested case-control study. AB - OBJECTIVES: This nested case-control study examines mortality from suicide in relation to estimated exposure to extremely low frequency electromagnetic fields (EMFs) in a cohort of 138,905 male electric utility workers. METHODS: Case control sampling included 536 deaths from suicide and 5348 eligible controls. Exposure was classified based on work in the most common jobs with increased exposure to magnetic fields and indices of cumulative exposure to magnetic fields based on a measurement survey. RESULTS: Suicide mortality was increased relative to work in exposed jobs and with indices of exposure to magnetic fields. Increased odds ratios (ORs) were found for years of employment as an electrician (OR 2.18; 95% confidence interval (95% CI) 1.25 to 3.80) or lineman (OR 1.59; 95% CI 1.18 to 2.14), whereas a decreased OR was found for power plant operators (OR 0.67; 95% CI 0.33 to 1.40). A dose response gradient with exposure to magnetic fields was found for exposure in the previous year, with a mortality OR of 1.70 (95% CI 1.00 to 2.90) in the highest exposure category. Stronger associations, with ORs in the range of 2.12-3.62, were found for men <50 years of age. CONCLUSION: These data provide evidence for an association between occupational electromagnetic fields and suicide that warrants further evaluation. A plausible mechanism related to melatonin and depression provides a direction for additional laboratory research as well as epidemiological evaluation. PMID- 10810114 TI - Symptoms, airway responsiveness, and exposure to dust in beech and oak wood workers. AB - OBJECTIVES: To investigate the relation between levels of cumulative exposure to wood dust and respiratory symptoms and the occurrence of bronchial hyperresponsiveness among beech and oak workers. METHODS: 114 Male woodworkers from five furniture factories and 13 male unexposed controls were examined. The unexposed control group was supplemented by 200 male historical controls. Statistical analyses were performed excluding and including the historical controls. Dust concentration was measured by personal sampling methods. Cumulative exposure to dust was calculated for each woodworker by multiplying the duration of the work by the intensity of exposure (years. mg/m(3)). Bronchial hyperresponsiveness was assessed by the methacholine bronchial challenge test. Subjects were labelled methacholine bronchial challenge positive if forced expiratory volume in 1 second (FEV(1)) fell by >/=20%. The linear dose-response slope was calculated as the last dose divided by the total dose given. RESULTS: 443 Dust samples were collected. The median cumulative exposure to dust was 110 years.mg/m(3) with lower and upper quartiles at 70 and 160 years.mg/m(3) Overall, no declines in FEV(1) and forced vital capacity (FVC) were found with increasing exposures. A dose-response relation was found between intensity of exposure on the one hand, and sore throat, increased prevalence of positive methacholine bronchial challenge tests, and steeper dose-response slope, on the other. CONCLUSION: Exposure to oak and beech dust may lead to the development of sore throat and bronchial hyperresponsiveness. PMID- 10810115 TI - Increased nitric oxide in exhaled air: an early marker of asthma in non-smoking aluminium potroom workers? AB - OBJECTIVES: To study exhaled nitric oxide (NO) as a marker of airway inflammation caused by potroom exposure, hypothesising that (a) workers exposed to potroom pollutants would have higher concentrations of NO in expired air than control subjects employed at the same plant but working outside of the potroom atmosphere, and (b) that concentrations of exhaled NO in potroom employees might be positively associated with concentrations of fluoride and exposure to dust. METHODS: A study group comprising 186 male subjects (aged 24-63 years), employed in the potrooms of one Norwegian aluminium smelter, and 40 comparable control subjects (aged 25-60 years) recruited from the same plant, were examined by measurements of exhaled and nasal concentrations of NO, spirometry, and a questionnaire on respiratory symptoms as a part of an annual health surveillance programme. Estimates of exposure to fluorides and dust for selected job categories were obtained by means of personal samplers carried by the workers. RESULTS: In the non-smokers, the concentrations of exhaled NO were higher in the potroom workers than in the controls (median (interquartile range) 9.3 (6.2-15.6) v 5.7 (4.6-8.3) ppb, p=0.001). The two groups did not differ in spirometry and asthma-like symptoms. Non-smoking potroom workers with asthma-like symptoms had higher concentrations of exhaled NO than those with no symptoms (median (interquartile range) 21.0 (19.3-41. 4) v 8.5 (5.9-12.8) ppb, p=0.001), but had comparable spirometric values. In subjects who smoked, the concentrations of exhaled NO did not differ significantly between potroom workers and controls (median (interquartile range) 4.6 (3.3-8.0) v 4.0 (3.4-5.1) ppb. Exhaled NO was not significantly associated with either duration of employment or routine measurements of dust and fluorides. CONCLUSIONS: Exposure to potroom pollutants is associated with increased concentrations of exhaled NO in non-smoking subjects. Nitric oxide in exhaled air may be an early marker of airway inflammation in aluminium potroom workers. PMID- 10810116 TI - Respiratory symptoms and lung function in alumina refinery employees. AB - OBJECTIVES: Employees in alumina refineries are known to be exposed to a number of potential respiratory irritants, particularly caustic mist and bauxite and alumina dusts. To examine the prevalence of work related respiratory symptoms and lung function in alumina refinery employees and relate these to their jobs. METHODS: 2964 current employees of three alumina refineries in Western Australia were invited to participate in a cross sectional study, and 89% responded. Subjects were given a questionnaire on respiratory symptoms, smoking, and occupations with additional questions on temporal relations between respiratory symptoms and work. Forced expiratory volume in 1 second (FEV(1)) and forced vital capacity (FVC) were measured with a rolling seal spirometer. Atopy was assessed with prick skin tests for common allergens. Associations between work and symptoms were assessed with Cox's regression to estimate prevalence ratios, and between work and lung function with linear regression. RESULTS: Work related wheeze, chest tightness, shortness of breath, and rhinitis were reported by 5.0%, 3.5%, 2.5%, and 9.5% of participants respectively. After adjustment for age, smoking, and atopy, most groups of production employees reported a greater prevalence of work related symptoms than did office employees. After adjustment for age, smoking, height, and atopy, subjects reporting work related wheeze, chest tightness, and shortness of breath had significantly lower mean levels of FEV(1) (186, 162, and 272 ml respectively) than subjects without these symptoms. Prevalence of most work related symptoms was higher at refinery 2 than at the other two refineries, but subjects at this refinery had an adjusted mean FEV(1) >60 ml higher than the others. Significant differences in FVC and FEV(1)/FVC ratio, but not FEV(1), were found between different process groups. CONCLUSIONS: There were significant differences in work related symptoms and lung function between process groups and refineries, but these were mostly not consistent. Undefined selection factors and underlying population differences may account for some of these findings but workplace exposures may also contribute. The differences identified between groups were unlikely to be clinically of note. PMID- 10810117 TI - Key Topics in Respiratory Medicine. PMID- 10810118 TI - Acute respiratory distress syndrome: a comprehensive clinical approach. PMID- 10810120 TI - Air pollutants and the respiratory tract. PMID- 10810119 TI - Microbiology in clinical practice, 3rd edition. PMID- 10810122 TI - Indoor air pollution and health. PMID- 10810121 TI - Rheumatic Diseases and the Environment. PMID- 10810123 TI - Chemical incident management for public health physicians. PMID- 10810124 TI - Strategy for prevention and control of the risks due to noise. AB - OBJECTIVES: To propose a strategy for progressively controlling the exposure to noise in industry as much as possible. To propose a method that could, in the first stage, be used by the workers and management themselves to control exposures to noise as much as possible, and then, in later stages, when necessary, progressively call in the assistance of specialists and experts to identify more complex solutions and organise personal protection and medical surveillance. METHODS: The strategy includes three stages. Stage 1 is observation, simple and easy to use by the workers to recognise the problems, identify straightforward solutions, and call for assistance when needed. Stage 2 is analysis, more complex but more costly, performed with the assistance of occupational health specialists to identify more technical control measures and set up a programme to conserve hearing. Stage 3 is expertise, performed with the assistance of acoustic experts for special measurements and control measures. CONCLUSIONS: The proposed strategy enriches the assessment procedure that is usually recommended, by providing for one preliminary stage used by the people directly concerned. It explicitly recognises (a) the competence of the workers and management about their working conditions and (b) that knowledge and measurements of acoustics are not an absolute prerequisite for solving-at least partly-noise problems. It attempts to organise in sequence and optimise the cooperation between the workers, the occupational health specialists, and the experts in acoustics. PMID- 10810125 TI - Influence of genetic susceptibility on the urinary excretion of 8 hydroxydeoxyguanosine of firefighters. AB - OBJECTIVES: Oxidative DNA damage has been implicated in carcinogenesis. The DNA damage can be assessed from the urinary excretion of the DNA-repair product 8 hydroxydeoxyguanosine (8-OH-dG). The factors were investigated that influenced the excretion of urinary 8-OH-dG in 78 firefighters. METHODS: 53 Out of 78 firefighters were exposed to fire within 5 days of the study and 25 were not. 8 OH-dG was measured by ELISA and the distribution of the genotypes of CYP1A1, CYP2E1, GSTM1, and GSTT1 was measured by polymerase chain reaction. RESULTS: The homozygous wild type frequencies of CYP1A1 MspI, CYP1A1 ile-val, CYP2E1, GSTM1, and GSTT1 were 31.5%, 56.2%, 60.3%, 50.7%, and 53.4%, respectively. The geometric mean of urinary 8-OH-dG was 14.1 ng/mg creatinine in more active firefighters and 12.3 ng/mg creatinine in non-exposed and less active subjects. Significantly increased concentrations of urinary 8-OH-dG were found to be associated with cigarette smoking, and 14% of the variation of 8-OH-dG was explained by cigarettes smoked per day. The CYP1A1 MspI, CYP1A1 ile-val, GSTM1, and GSTT1 genetic polymorphisms were not found to be significantly associated with the urinary excretion of 8-OH-dG. However, the subjects carrying the CYP2E1 mutant type excreted higher concentrations of 8-OH-dG and there was a marginally significant interaction of GSTT1 with firefighting activity. Multiple regression analysis confirmed that smoking was the strongest predictor of excretion of 8-OH dG. Age, body mass index, and firefighting activity were not significant predictive factors for urinary 8-OH-dG. CONCLUSION: Smoking and CYP2E1 gene polymorphism may be important factors in carcinogenesis and the GSTT1 positive genotype may be a genetic susceptibility factor in firefighters who are exposed regularly to various chemical carcinogens. PMID- 10810126 TI - Occupational exposure to formaldehyde and wood dust and nasopharyngeal carcinoma. AB - OBJECTIVES: To investigate whether occupational exposures to formaldehyde and wood dust increase the risk of nasopharyngeal cancer (NPC). METHODS: A multicentered, population based case-control study was carried out at five cancer registries in the United States participating in the National Cancer Institute's SEER program. Cases (n=196) with a newly diagnosed NPC between 1987 and 1993, and controls (n=244) selected over the same period from the general population through random digit dialing participated in structured telephone interviews which inquired about suspected risk factors for the disease, including a lifetime history of occupational and chemical exposure. Histological type of cancer was abstracted from clinical records of the registries. Potential exposure to formaldehyde and wood dust was assessed on a job by job basis by experienced industrial hygienists who were blinded as to case or control status. RESULTS: For formaldehyde, after adjusting for cigarette use, race, and other risk factors, a trend of increasing risk of squamous and unspecified epithelial carcinomas was found for increasing duration (p=0.014) and cumulative exposure (p=0.033) but not for maximum exposure concentration. The odds ratio (OR) for people cumulatively exposed to >1.10 ppm-years was 3.0 (95% confidence interval (95% CI) 1.3 to 6.6) compared with those considered unexposed. In analyses limited to jobs considered definitely exposed, these trends became stronger. The associations were most evident among cigarette smokers. By contrast, there was no association between potential exposure to formaldehyde and undifferentiated and non-keratinising carcinomas. There was little evidence that exposure to wood dust increased risk of NPC, as modest crude associations essentially disappeared after control for potential exposure to formaldehyde. CONCLUSIONS: These results support the hypothesis that occupational exposure to formaldehyde, but not wood dust, increases risk of NPC. This association seems to be specific to squamous cell carcinomas. Established cohorts of workers exposed to formaldehyde and wood dust should continue to be monitored for NPC and other respiratory cancers. Future studies of NPC should take into account histological type in assessing risk from environmental and host factors. PMID- 10810127 TI - Lung cancer in Yorkshire chrome platers, 1972-97. AB - OBJECTIVES: To investigate mortality from lung cancer in chrome platers, a group exposed to chromic acid. METHODS: The mortality of a cohort of 1087 chrome platers (920 men, 167 women) from 54 plants situated in the West Riding of Yorkshire, United Kingdom, was investigated for the period 1972-97. All subjects were employed as chrome platers for >/=3 months and all were alive on 31 May 1972. Mortality data were also available for a cohort of 1163 comparison workers with no known occupational exposure to chrome compounds (989 men, 174 women). Information on duration of chrome work and smoking habits collected for a cross sectional survey carried out in 1969-72 were available for 916 (84.3%) of the chrome platers; smoking habits were available for 1004 (86.3%) comparison workers. Two analytical approaches were used, indirect standardisation and Poisson regression. RESULTS: Based on serial mortality rates for the general population of England and Wales, significantly increased mortality from lung cancer was observed (obs) in male chrome platers (obs 60, expected (exp) 32.5, standardised mortality ratio (SMR) 185, p<0. 001) but not in male comparison workers (obs 47, exp 36.9, SMR 127). Positive trends were not shown for duration of employment exposed to chrome, although data on working after 1972 were not available. CONCLUSIONS: Confident interpretation is not possible but occupational exposures to hexavalent chromium may well have been involved in the increased mortality from lung cancer found in this cohort of chrome platers. PMID- 10810128 TI - Lung function over the first 3 years of a professional diving career. AB - OBJECTIVES: To characterise diving exposure and pulmonary function in professional divers at the start of their formal education and during the first 3 years of their professional career. METHODS: The study included 87 men at the start of their education as professional divers. At follow up 1 and 3 years after the school 83 and 81 divers respectively were re-examined. Assessment of lung function included dynamic lung volumes and flows and transfer factor for carbon monoxide (Tl(CO)). RESULTS: 69 Divers had preschool SCUBA diving experience and had a median number of 70 dives (range 2-3000) to a median maximal depth of 40 (range 10-73) metres. During the 15 week introductory diving course, they had 44 dives (range 38-50) in the depth range 10-50 metres. The median number of dives over the follow up period was 95 (range 0-722) to a maximal median depth of 38 (range 0-98) metres. At the start of the diving course there were no differences in forced vital capacity (FVC), forced expired volume in 1 second (FEV(1)), and in Tl(CO) between the 69 pre-exposed divers and the 18 never exposed divers. The FVC was significantly larger than predicted in both groups. At follow up at 3 years there was a significant reduction in mean (SD) FEV(1) of 1.8% (6.5), in forced mid-expiratory flow rate (FEF(25-75%)) of 6.5% (11.7) and in forced expiratory flow at 75% of FVC expired (FEF(75%)) of 10.4% (16. 8). There was no change in FVC. The Tl(CO) was significantly decreased by 4.6% (8.8). No significant effects were found of cumulative diving exposure, including the number of dives, on the relative changes of any of the lung function variables. CONCLUSIONS: The results indicate that divers initially belong to a selected group with large FVC. Exposure to diving may contribute to changes in pulmonary function, mostly affecting small airways conductance. PMID- 10810129 TI - Exposure biomarkers and risk from gluing and heating of polyurethane: a cross sectional study of respiratory symptoms. AB - OBJECTIVES: To define the relation between exposure to polyurethane (PUR) glue, biomarkers of exposure and effect, and work related symptoms that occur at least once a week. METHODS: In a cross sectional study, 152 workers and 14 clerks in a factory with exposure to sprayed and heated PUR glue containing 4, 4' diphenylmethane (MDI) or 1,6-hexamethylene (HDI) di-isocyanate were examined with gas chromatography-mass spectrometry (GC-MS) for metabolites of MDI in plasma (P MDX) and urine (U-MDX), 2,4- and 2, 6-toluene di-isocyanate (TDI; P-TDX, U-TDX) and HDI in plasma and urine, specific serum IgG (S-IgG-MDI, S-IgG-HDI, and S-IgG TDI, respectively) and IgE (S-IgE-MDI). Work related symptoms of the eyes and airways (nose or lower airways, or both), and lung function were also evaluated. RESULTS: P-MDX was detected in 65% of the workers, U-TDX in 47%, HDX in none. Three per cent were positive for S-IgE-MDI, 33% for S-IgG-MDI, 32% for S-IgG-TDI, and 12% for S-IgG-HDI. A few clerks had metabolites, and some had antibodies. Most metabolites and immunoglobulins were slightly correlated-for example, P-MDX v S-IgG-MDI: r(s)=0.21. Workers who heated glue had increased P-MDX (odds ratio (OR)=12 for a value above the median) and S-IgG-MDI (OR=3.7), sprayers P-2,4-TDX (OR=6.2) and P-2,6-TDX (OR=16). Twenty six per cent of the workers had work related symptoms of the airways, 21% from the nose, 11% from the lower airways. Spraying of glue increased the risk of work related symptoms and slightly decreased lung function. U-MDX was associated with work related symptoms from the airways (OR=3.7) and P-2,6-TDX with work related symptoms from the lower airways (OR=6.6). S-IgG-MDI was related to work related symptoms from the airways (OR=2.6). CONCLUSIONS: There were relations between exposures to sprayed and heated PUR glue based on MDI and HDI, concentrations of metabolites of MDI and TDI in plasma and urine, specific IgG serum antibodies against MDI, TDI, and HDI, and work related symptoms. PMID- 10810130 TI - Exposure of Paris taxi drivers to automobile air pollutants within their vehicles. AB - OBJECTIVES: To study the exposure of Parisian taxi drivers to automobile air pollutants during their professional activity. METHODS: A cross sectional study was carried out from 27 January to 27 March 1997, with measurements performed in the vehicles of 29 randomly selected drivers. Carbon monoxide (CO) content was measured over an 8 hour period by a CO portable monitor. The fine suspended particles were measured according to the black smoke index (BS), with a flow controlled portable pump provided with a cellulose filter. The nitrogen oxides, NO and NO(2) were measured with a passive sampler. RESULTS: These drivers are exposed during their professional activity to relatively high concentrations of pollutants (mean, median (SD) 3.8, 2 (1.7) ppm for CO, 168, 164 (53) micrograms/m(3) for BS, 625, 598 (224) micrograms/m(3) for NO, and 139, 131 (43) micrograms/m(3) for NO(2).) For CO the concentrations were clearly lower than the threshold values recommended by the World Health Organisation. The situation is less satisfactory for the other pollutants, especially for the BS index. All concentrations of pollutants recorded were noticeably higher than concentrations in air recorded by the ambient Parisian air monitoring network and were close to, or slightly exceeded, the concentrations measured at the fixed stations close to automobile traffic. Pollutant concentrations were also influenced greatly by weather conditions. CONCLUSION: This first French study conducted in taxi drivers shows that they are highly exposed to automobile pollutants. The results would justify a medical follow up of this occupational group. PMID- 10810131 TI - Update of a study of crude oil production workers 1946-94. AB - OBJECTIVE: To update information on workers in the petroleum industry engaged in the production of crude oil to determine whether the patterns of mortality have changed with 14 additional years of follow up. METHODS: All workers were employed at company production and pipeline locations sometime during 1946-94. The cohort now consists of 24 124 employees with an average of 22 years of follow up. RESULTS: The overall mortality, and most cause specific mortalities were lower than or similar to those for the general United States population. For white men (81% of the cohort), there were 4361 observed deaths and 5945 expected, resulting in a significantly lower standardised mortality ratio (SMR) of 73. There were significant deficits for all the leading causes of death in the United States including all cancers, cancer of the lung, stroke, heart disease, respiratory disease, and accidents. Slightly increased mortality was found for cancer of the prostate, cancer of the brain and central nervous system, and cancer of other lymphatic tissue. For benign and unspecified neoplasms, the SMR was 152 (95% confidence interval (95% CI) 95 to 230). There was a significant increase for acute myelogenous leukaemia that was restricted to people who were first employed before 1940 and who were employed in production and pipeline jobs for >30 years. Overall mortality patterns for non-white men and women were similar to those for white men. Mortality patterns for white men were also examined by duration of employment, time first employed, and by job group. CONCLUSIONS: The results of the updated study showed a favourable mortality experience for crude oil production workers compared with the United States population. PMID- 10810132 TI - Mortality from multiple sclerosis and exposure to residential and occupational solar radiation: a case-control study based on death certificates. AB - OBJECTIVES: To explore whether mortality from multiple sclerosis is negatively associated with exposure to sunlight. METHODS: Two case-control studies based on death certificates were conducted for mortality from multiple sclerosis and non melanoma skin cancer (as a positive control) to examine associations with residential and occupational exposure to sunlight. Cases were all deaths from multiple sclerosis between 1984 and 1995 in 24 states of the United States. Controls, which were age frequency matched to a series of cases, excluded cancer and certain neurological deaths. The effects of occupational exposure to sunlight were assessed among subjects with usual occupations requiring substantial activity, so as to exclude those whose indoor jobs resulted from disabilities subsequent to the onset of the disease. Multiple logistic regression analyses were applied, with adjustment for age, sex, race, and socioeconomic status. RESULTS: Unlike mortality from skin cancer, mortality from multiple sclerosis was negatively associated with residential exposure to sunlight (odds ratio (OR)=0.53 (multiple sclerosis) and OR=1.24 (skin cancer)). Odds ratios for the highest occupational exposure to sunlight were 0.74 (95% confidence interval (95% CI) 0.61 to 0.89) for mortality from multiple sclerosis, compared with 1.21 (1.09 to 1.34) for mortality from non-melanoma skin cancer. The OR was 0.24 for the combined effect of the highest levels of residential and occupational exposure to sunlight on multiple sclerosis, compared with an OR of 1.38 for skin cancer. CONCLUSIONS: In this exploratory study, mortality from multiple sclerosis, unlike mortality from skin cancer, was negatively associated with both residential and occupational exposure to sunlight. PMID- 10810133 TI - Acute vascular responses to the frequency of vibration transmitted to the hand. AB - OBJECTIVES: To investigate the acute effects of the frequency of hand transmitted vibration on finger circulation. A further aim was to investigate whether the frequency weighting assumed in current standards for hand transmitted vibration reflects the haemodynamic changes which occur in the fingers exposed to vibration with different frequencies but with the same frequency weighted acceleration magnitude. METHODS: Finger skin temperature (FST) and finger blood flow (FBF) were measured in the middle fingers of both hands of 10 healthy men. With a static load of 10 N, the right hand was exposed for 15 minutes to the following root mean square (rms) acceleration magnitudes and frequencies of vertical vibration: 5.5 m/s(2) at 16 Hz; 11 m/s(2) at 31.5 Hz; 22 m/s(2) at 63 Hz; 44 m/s(2) at 125 Hz; and 88 m/s(2) at 250 Hz. These exposures to vibration produce the same frequency weighted acceleration magnitude (5.5 m/s(2) rms) according to the frequency weighting included in the international standard ISO 5349. A control condition consisted of exposure to the static load only. Finger circulation was measured before application of the vibration and static load and at fixed intervals during exposure to vibration and a 45 minute recovery period. RESULTS: No significant changes in finger circulation were found with only the static load. The FST did not change significantly during or after acute exposure to vibration. In the vibrated right finger, exposures to vibration with frequencies of 31. 5-250 Hz provoked a greater reduction in FBF than did vibration of 16 Hz or the static load only. In the non-vibrated left finger, the FBF measured with vibration at each frequency of 63-250 Hz was significantly lower than that measured with static load only. The reduction in FBF during exposure to vibration with any frequency was stronger in the vibrated finger than in the non-vibrated finger. In both fingers, there was a progressive decrease in FBF after the end of exposure to vibration with frequencies of 31.5-250 Hz. The higher the frequency of vibration, the stronger the decrease in FBF in both fingers during recovery. CONCLUSIONS: Acute exposures to vibration with equal frequency weighted magnitude reduce the FBF in both vibrated and non-vibrated fingers for frequencies between 31.5 and 250 Hz. The extent of digital vasoconstriction after exposure to vibration increases with increasing frequency. The frequency weighting given in current standards tends to overestimate the vasoconstriction associated with acute exposures to vibration frequencies around 16 Hz. PMID- 10810134 TI - Work-related musculoskeletal disorders: report, workshop summary, and workshop papers PMID- 10810135 TI - Occupational musculoskeletal disorders, second edition PMID- 10810136 TI - Air pollution and health PMID- 10810138 TI - Microarrays: determining the balance of cellular transcription. PMID- 10810140 TI - Genome relationships: the grass model in current research. PMID- 10810139 TI - Comparative genome organization in plants: from sequence and markers to chromatin and chromosomes. PMID- 10810141 TI - Identification of the SAAT gene involved in strawberry flavor biogenesis by use of DNA microarrays. AB - Fruit flavor is a result of a complex mixture of numerous compounds. The formation of these compounds is closely correlated with the metabolic changes occurring during fruit maturation. Here, we describe the use of DNA microarrays and appropriate statistical analyses to dissect a complex developmental process. In doing so, we have identified a novel strawberry alcohol acyltransferase (SAAT) gene that plays a crucial role in flavor biogenesis in ripening fruit. Volatile esters are quantitatively and qualitatively the most important compounds providing fruity odors. Biochemical evidence for involvement of the SAAT gene in formation of fruity esters is provided by characterizing the recombinant protein expressed in Escherichia coli. The SAAT enzyme showed maximum activity with aliphatic medium-chain alcohols, whose corresponding esters are major components of strawberry volatiles. The enzyme was capable of utilizing short- and medium chain, branched, and aromatic acyl-CoA molecules as cosubstrates. The results suggest that the formation of volatile esters in fruit is subject to the availability of acyl-CoA molecules and alcohol substrates and is dictated by the temporal expression pattern of the SAAT gene(s) and substrate specificity of the SAAT enzyme(s). PMID- 10810142 TI - Members of the Arabidopsis HRT/RPP8 family of resistance genes confer resistance to both viral and oomycete pathogens. AB - Turnip crinkle virus (TCV) inoculation onto TCV-resistant Arabidopsis leads to a hypersensitive response (HR) controlled by the dominant gene HRT. HRT is a member of the class of resistance (R) genes that contain a leucine zipper, a nucleotide binding site, and leucine-rich repeats. The chromosomal position of HRT and its homology to resistance gene RPP8 and two RPP8 homologs indicate that unequal crossing over and gene conversion may have contributed to HRT evolution. RPP8 confers resistance to an oomycete pathogen, Peronospora parasitica. Despite very strong similarities within the HRT/RPP8 family, HRT and RPP8 are specific for the respective pathogens they detect. Hence, the HRT/RPP8 family provides molecular evidence that sequence changes between closely related members of multigene families can generate novel specificities for radically different pathogens. Transgenic plants expressing HRT developed an HR but generally remained susceptible to TCV because of a second gene, RRT, that regulates resistance to TCV. However, several transgenic plants that overexpressed HRT produced micro-HRs or no HR when inoculated with TCV and were resistant to infection. Expression of the TCV coat protein gene in seedlings containing HRT resulted in massive necrosis and death, indicating that the avirulence factor detected by the HRT encoded protein is the TCV coat protein. PMID- 10810143 TI - Resistance to turnip crinkle virus in Arabidopsis is regulated by two host genes and is salicylic acid dependent but NPR1, ethylene, and jasmonate independent. AB - Inoculation of turnip crinkle virus (TCV) on the resistant Arabidopsis ecotype Dijon (Di-17) results in the development of a hypersensitive response (HR) on the inoculated leaves. To assess the role of the recently cloned HRT gene in conferring resistance, we monitored both HR and resistance (lack of viral spread to systemic tissues) in the progeny of a cross between resistant Di-17 and susceptible Columbia plants. As expected, HR development segregated as a dominant trait that corresponded with the presence of HRT. However, all of the F(1) plants and three-fourths of HR(+) F(2) plants were susceptible to the virus. These results suggest the presence of a second gene, termed RRT, that regulates resistance to TCV. The allele present in Di-17 appears to be recessive to the allele or alleles present in TCV-susceptible ecotypes. We also demonstrate that HR formation and TCV resistance are dependent on salicylic acid but not on ethylene or jasmonic acid. Furthermore, these phenomena are unaffected by mutations in NPR1. Thus, TCV resistance requires a yet undefined salicylic acid dependent, NPR1-independent signaling pathway. PMID- 10810144 TI - Virus-induced silencing of a plant cellulose synthase gene. AB - Specific cDNA fragments corresponding to putative cellulose synthase genes (CesA) were inserted into potato virus X vectors for functional analysis in Nicotiana benthamiana by using virus-induced gene silencing. Plants infected with one group of cDNAs had much shorter internode lengths, small leaves, and a "dwarf" phenotype. Consistent with a loss of cell wall cellulose, abnormally large and in many cases spherical cells ballooned from the undersurfaces of leaves, particularly in regions adjacent to vascular tissues. Linkage analyses of wall polysaccharides prepared from infected leaves revealed a 25% decrease in cellulose content. Transcript levels for at least one member of the CesA cellulose synthase gene family were lower in infected plants. The decrease in cellulose content in cell walls was offset by an increase in homogalacturonan, in which the degree of esterification of carboxyl groups decreased from approximately 50 to approximately 33%. The results suggest that feedback loops interconnect the cellular machinery controlling cellulose and pectin biosynthesis. On the basis of the phenotypic features of the infected plants, changes in wall composition, and the reduced abundance of CesA mRNA, we concluded that the cDNA fragments silenced one or more cellulose synthase genes. PMID- 10810145 TI - Differential gene expression in response to mechanical wounding and insect feeding in Arabidopsis. AB - Wounding in multicellular eukaryotes results in marked changes in gene expression that contribute to tissue defense and repair. Using a cDNA microarray technique, we analyzed the timing, dynamics, and regulation of the expression of 150 genes in mechanically wounded leaves of Arabidopsis. Temporal accumulation of a group of transcripts was correlated with the appearance of oxylipin signals of the jasmonate family. Analysis of the coronatine-insensitive coi1-1 Arabidopsis mutant that is also insensitive to jasmonate allowed us to identify a large number of COI1-dependent and COI1-independent wound-inducible genes. Water stress was found to contribute to the regulation of an unexpectedly large fraction of these genes. Comparing the results of mechanical wounding with damage by feeding larvae of the cabbage butterfly (Pieris rapae) resulted in very different transcript profiles. One gene was specifically induced by insect feeding but not by wounding; moreover, there was a relative lack of water stress-induced gene expression during insect feeding. These results help reveal a feeding strategy of P. rapae that may minimize the activation of a subset of water stress-inducible, defense-related genes. PMID- 10810146 TI - Transgenic Arabidopsis plants expressing a fungal cutinase show alterations in the structure and properties of the cuticle and postgenital organ fusions. AB - A major structural component of the cuticle of plants is cutin. Analysis of the function of cutin in vivo has been limited because no mutants with specific defects in cutin have been characterized. Therefore, transgenic Arabidopsis plants were generated that express and secrete a cutinase from Fusarium solani f sp pisi. Arabidopsis plants expressing the cutinase in the extracellular space showed an altered ultrastructure of the cuticle and an enhanced permeability of the cuticle to solutes. In addition, pollen could germinate on fully differentiated leaves of cutinase-expressing plants but not on control leaves. These differences coincided with strong postgenital organ fusions. The junctions of the fusions contained pectic polysaccharides. As fused organs grew apart from each other, organ deformations and protrusions of epidermal cells developed at positions with high mechanical stress. These results demonstrate that an intact cutin layer not only is important for plant-environment interactions but also prevents fusions between different plant organs and is therefore necessary for normal epidermal differentiation and organ formation. PMID- 10810147 TI - Protein recycling from the Golgi apparatus to the endoplasmic reticulum in plants and its minor contribution to calreticulin retention. AB - Using pulse-chase experiments combined with immunoprecipitation and N-glycan structural analysis, we showed that the retrieval mechanism of proteins from post endoplasmic reticulum (post-ER) compartments is active in plant cells at levels similar to those described previously for animal cells. For instance, recycling from the Golgi apparatus back to the ER is sufficient to block the secretion of as much as 90% of an extracellular protein such as the cell wall invertase fused with an HDEL C-terminal tetrapeptide. Likewise, recycling can sustain fast retrograde transport of Golgi enzymes into the ER in the presence of brefeldin A. However, on the basis of our data, we propose that this retrieval mechanism in plants has little impact on the ER retention of a soluble ER protein such as calreticulin. Indeed, the latter is retained in the ER without any N-glycan related evidence for a recycling through the Golgi apparatus. Taken together, these results indicate that calreticulin and perhaps other plant reticuloplasmins are possibly largely excluded from vesicles exported from the ER. Instead, they are probably retained in the ER by mechanisms that rely primarily on signals other than H/KDEL motifs. PMID- 10810148 TI - The NPH4 locus encodes the auxin response factor ARF7, a conditional regulator of differential growth in aerial Arabidopsis tissue. AB - Organ bending through differential growth represents a major mechanism by which plants are able to adaptively alter their morphology in response to local changes in the environment. Two plant hormones, auxin and ethylene, have been implicated as regulators of differential growth responses; however, the mechanisms by which they elicit their effects remain largely unknown. Here, we describe isolation of the NPH4 gene of Arabidopsis, which is conditionally required for differential growth responses of aerial tissues, and we report that NPH4 encodes the auxin regulated transcriptional activator ARF7. The phenotypes of nph4 mutants, which include multiple differential growth defects associated with reduced auxin responsiveness, including impaired auxin-induced gene expression, are consistent with the predicted loss of function of a transcriptional activator, and these phenotypes indicate that auxin-dependent changes in gene transcription are prerequisite for proper organ bending responses. Although NPH4/ARF7 appears to be a major regulator of differential growth, it is not the sole regulator because phenotypes of nph4 null mutants were suppressed by application of ethylene. This latter finding illustrates the intimate connection between auxin and ethylene in the control of growth in higher plants. PMID- 10810149 TI - Pti4 is induced by ethylene and salicylic acid, and its product is phosphorylated by the Pto kinase. AB - The tomato Pti4 gene encodes a transcription factor that was identified on the basis of its specific interaction with the product of the Pto disease resistance gene in a yeast two-hybrid system. We show here that the Pti4 protein specifically binds the GCC-box cis element, which is present in the promoter region of many pathogenesis-related (PR) genes. Expression of the Pti4 gene in tomato leaves was rapidly induced by ethylene and by infection with Pseudomonas syringae pv tomato, and this induction preceded expression of GCC-box-containing PR genes. Although salicylic acid also induced Pti4 gene expression, it did not induce GCC-box PR genes. Rather, salicylic acid antagonized ethylene-mediated expression of GCC-box PR genes. We demonstrate that the Pti4 protein is specifically phosphorylated by the Pto kinase and that this phosphorylation enhances binding of Pti4 to the GCC box. In addition, induced overexpression of Pto and Pti4 in tomato leaves resulted in a concomitant increase in GCC-box PR genes. Our results support a model in which phosphorylation of the Pti4 protein by the Pto kinase enhances the ability of Pti4 to activate expression of GCC-box PR genes in tomato. PMID- 10810150 TI - Identification, purification, and molecular cloning of a putative plastidic glucose translocator. AB - During photosynthesis, part of the fixed carbon is directed into the synthesis of transitory starch, which serves as an intermediate carbon storage facility in chloroplasts. This transitory starch is mobilized during the night. Increasing evidence indicates that the main route of starch breakdown proceeds by way of hydrolytic enzymes and results in glucose formation. This pathway requires a glucose translocator to mediate the export of glucose from the chloroplasts. We have reexamined the kinetic properties of the plastidic glucose translocator and, using a differential labeling procedure, have identified the glucose translocator as a component of the inner envelope membrane. Peptide sequence information derived from this protein was used to isolate cDNA clones encoding a putative plastidic glucose translocator from spinach, potato, tobacco, Arabidopsis, and maize. We also present the molecular characterization of a candidate for a hexose transporter of the plastid envelope membrane. This transporter, initially characterized more than 20 years ago, is closely related to the mammalian glucose transporter GLUT family and differs from all other plant hexose transporters that have been characterized to date. PMID- 10810151 TI - Resistance gene-dependent activation of a calcium-dependent protein kinase in the plant defense response. AB - In the Cf-9/Avr9 gene-for-gene interaction, the Cf-9 resistance gene from tomato confers resistance to the fungal pathogen Cladosporium fulvum, which expresses the corresponding pathogen-derived avirulence product Avr9. To understand R gene function and dissect the signaling mechanisms involved in the induction of plant defenses, we studied Cf-9/Avr9-dependent activation of protein kinases in transgenic Cf9 tobacco cell cultures. Using a modified in-gel kinase assay with histone as substrate, we identified a membrane-bound, calcium-dependent protein kinase (CDPK) that showed a shift in electrophoretic mobility from 68 to 70 kD within 5 min after Avr9 elicitor was added. This transition from the nonelicited to the elicited CDPK form was caused by a phosphorylation event and was verified when antibodies to CDPK were used for protein gel blot analysis. In addition, the interconversion of the corresponding CDPK forms could be induced in vitro in both directions by treatment with either phosphatase or ATP. In vitro protein kinase activity toward syntide-2 or histone with membrane extracts or gel-purified enzyme was dependent on Ca(2)+ content and was compromised by the calmodulin antagonist N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) but not by its inactive isoform N-(6-aminohexyl)-1-naphthalenesulfonamide. In these assays, the CDPK activity in elicited samples, reflecting predominantly the phosphorylated 70-kD CDPK form, was greater than in nonelicited samples. Thus, Avr9/Cf-9-dependent phosphorylation and subsequent transition from the nonelicited to the elicited form correlate with the activation of a CDPK isoform after in vivo stimulation. Because that transition was not inhibited by W-7, the in vivo CDPK activation probably is not the result of autophosphorylation. Studies with pharmacological inhibitors indicated that the identified CDPK is independent of or is located upstream from a signaling pathway that is required for the Avr9-induced active oxygen species. PMID- 10810152 TI - Differences in the recognition of glucan elicitor signals between rice and soybean: beta-glucan fragments from the rice blast disease fungus Pyricularia oryzae that elicit phytoalexin biosynthesis in suspension-cultured rice cells. AB - Partial acid/enzymatic hydrolysis of the beta-(1-->3, 1-->6)-glucan from the cell walls of the rice blast disease fungus Pyricularia oryzae (Magnaporthe grisea) released elicitor-active fragments that induced phytoalexin biosynthesis in suspension-cultured rice cells. From the digestion of the glucan by an endo-beta (1-->3)-glucanase, one highly elicitor-active glucopentaose was purified as a reduced compound, tetraglucosyl glucitol. The structure of this tetraglucosyl glucitol as well as two other related tetraglucosyl glucitols was elucidated as follows: (1) Glcbeta(1-->3)Glcbeta(1-->3)(Glcbeta(1-->6)) Glcbeta(1-->3)Glucitol (most active fragment); (2) Glcbeta(1-->3)(Glcbeta(1-->6))Glcbeta(1-->3)Glcbeta (1-->3)Glucitol; and (3) Glcbeta(1-->6) Glcbeta(1-->3)Glcbeta(1-->3)Glcbeta(1- >3)Glucitol. However, a synthetic hexa-beta-glucoside, known as a minimal structural element for the phytoalexin elicitor for soybean cotyledon cells, did not induce phytoalexin biosynthesis in the rice cells. Conversely, the beta glucan fragment from P. oryzae did not induce phytoalexin biosynthesis in the soybean cotyledon cells, indicating differences in the recognition of glucooligosaccharide elicitor signals in these two plants. Because rice cells have been shown to recognize chitin fragments larger than pentamers as potent elicitors, these results also indicate that the rice cells can recognize at least two types of oligosaccharides from fungal cell walls as signal molecules to initiate defense response. PMID- 10810153 TI - Deterministic features of side-chain main-chain hydrogen bonds in globular protein structures. AB - A total of 19 835 polar residues from a data set of 250 non-homologous and highly resolved protein crystal structures were used to identify side-chain main-chain (SC-MC) hydrogen bonds. The ratio of the number of SC-MC hydrogen bonds to the total number of polar residues is close to 1:2, indicating the ubiquitous nature of such hydrogen bonds. Close to 56% of the SC-MC hydrogen bonds are local involving side-chain acceptor/donor ('i') and a main-chain donor/acceptor within the window i-5 to i+5. These short-range hydrogen bonds form well defined conformational motifs characterized by specific combinations of backbone and side chain torsion angles. (a) The Ser/Thr residues show the greatest preference in forming intra-helical hydrogen bonds between the atoms O(gamma)(i) and O(i-4). More than half the examples of such hydrogen bonds are found at the middle of alpha-helices rather than at their ends. The most favoured motif of these examples is alpha(R)alpha(R)alpha(R)alpha(R)(g(-)). (b) These residues also show great preference to form hydrogen bonds between O(gamma)(i) and O(i-3), which are closely related to the previous type and though intra-helical, these hydrogen bonds are more often found at the C-termini of helices than at the middle. The motif represented by alpha(R)alpha(R)alpha(R)alpha(R)(g(+)) is most preferred in these cases. (c) The Ser, Thr and Glu are the most frequently found residues participating in intra-residue hydrogen bonds (between the side-chain and main chain of the same residue) which are characterized by specific motifs of the form beta(g(+)) for Ser/Thr residues and alpha(R)(g(-)g(+)t) for Glu/Gln. (d) The side chain acceptor atoms of Asn/Asp and Ser/Thr residues show high preference to form hydrogen bonds with acceptors two residues ahead in the chain, which are characterized by the motifs beta (tt')alphaR and beta(t)alpha(R), respectively. These hydrogen bonded segments, referred to as Asx turns, are known to provide stability to type I and type I' beta-turns. (e) Ser/Thr residues often form a combination of SC-MC hydrogen bonds, with the side-chain donor hydrogen bonded to the carbonyl oxygen of its own peptide backbone and the side-chain acceptor hydrogen bonded to an amide hydrogen three residues ahead in the sequence. Such motifs are quite often seen at the beginning of alpha-helices, which are characterized by the beta(g(+))alpha(R)alpha(R) motif. A remarkable majority of all these hydrogen bonds are buried from the protein surface, away from the surrounding solvent. This strongly indicates the possibility of side-chains playing the role of the backbone, in the protein interiors, to satisfy the potential hydrogen bonding sites and maintaining the network of hydrogen bonds which is crucial to the structure of the protein. PMID- 10810154 TI - Evaluation of protein-protein association energies by free energy perturbation calculations. AB - The association energy upon binding of different amino acids in the specificity pocket of trypsin was evaluated by free energy perturbation calculations on complexes between bovine trypsin (BT) and bovine pancreatic trypsin inhibitor (BPTI). Three simulations of mutations of the primary binding residue (P(1)) were performed (P(1)-Ala to Gly, P(1)-Met to Gly and P(1)-Met to Ala) and the resulting differences in association energy (DeltaDeltaG(a)) are 2. 28, 5.08 and 2.93 kcal/mol for P(1)-Ala to Gly, P(1)-Met to Gly and to Ala with experimental values of 1.71, 4.62 and 2.91 kcal/mol, respectively. The calculated binding free energy differences are hence in excellent agreement with the experimental binding free energies. The binding free energies, however, were shown to be highly dependent on water molecules at the protein-protein interface and could only be quantitatively estimated if the correct number of such water molecules was included. Furthermore, the cavities that were formed when a large amino acid side chain is perturbed to a smaller one seem to create instabilities in the systems and had to be refilled with water molecules in order to obtain reliable results. In addition, if the protein atoms that were perturbed away were not replaced by water molecules, the simulations dramatically overestimated the initial state of the free energy perturbations. PMID- 10810155 TI - Inversion of the roles of the nucleophile and acid/base catalysts in the covalent binding of epoxyalkyl xyloside inhibitor to the catalytic glutamates of endo-1,4 beta-xylanase (XYNII): a molecular dynamics study. AB - X-Ray crystal structures have revealed that 2, 3-epoxypropyl-beta-D-xyloside reacts with endo-1,4-beta-xylanase (XYNII) by forming a covalent bond with Glu86. In contrast, 3, 4-epoxybutyl-beta-D-xyloside forms a covalent bond with Glu177. In the normal enzyme reaction Glu86 acts as the catalytic nucleophile and Glu177 as the acid/base catalyst. To rationalize the observed reactivity of the two mechanism-based inhibitors, we carried out eight 300 ps molecular dynamics simulations for different enzyme-inhibitor complexes. Simulations were done for both stereo isomers (R and S) of the inhibitors and for enzyme in which the protonation state of the nucleophile and acid/base catalyst was normal (Glu86 charged, Glu177 neutral) and in which the roles of the catalytic residues were reversed (Glu86 neutral, Glu177 charged). The number of reactive conformations found in each simulation was used to predict the reactivity of epoxy inhibitors. The conformation was considered to be a reactive one when at the same time (i) the proton of the catalytic acid was close (<2.9/3.4/3.9 A) to the oxirane oxygen of the inhibitor, (ii) the nucleophile was close to the terminal carbon of the oxirane group (<3.4/3.9/4.4 A) and (iii) the nucleophile approached the terminal carbon from a reactive angle (<30/45/60 degrees from an ideal attack angle). On the basis of the number of reactive conformations, 2,3-epoxypropyl-beta-D xyloside was predicted to form a covalent bond with Glu86 and 3, 4-epoxybutyl beta-D-xyloside with Glu177, both in agreement with the experiment. Thus, the MD simulations and the X-ray structures indicate that in the covalent binding of 3, 4-epoxybutyl-beta-D-xyloside the roles of the catalytic glutamates of XYNII are reversed from that of the normal enzyme reaction. PMID- 10810156 TI - Crystal structures of 3-isopropylmalate dehydrogenases with mutations at the C terminus: crystallographic analyses of structure-stability relationships. AB - Thermal stability of the Thermus thermophilus isopropylmalate dehydrogenase enzyme was substantially lost upon the deletion of three residues from the C terminus. However, the stability was partly recovered by the addition of two, four and seven amino acid residues (called HD177, HD708 and HD711, respectively) to the C-terminal region of the truncated enzyme. Three structures of these mutant enzymes were determined by an X-ray diffraction method. All protein crystals belong to space group P2(1) and their structures were solved by a standard molecular replacement method where the original dimer structure of the A172L mutant was used as a search model. Thermal stability of these mutant enzymes is discussed based on the 3D structure with special attention to the width of the active-site groove and the minor groove, distortion of beta-sheet pillar structure and size of cavity in the domain-domain interface around the C terminus. Our previous studies revealed that the thermal stability of isopropylmalate dehydrogenase increases when the active-site cleft is closed (the closed form). In the present study it is shown that the active-site cleft can be regulated by open-close movement of the minor groove located at the opposite side to the active-site groove on the same subunit, through a paperclip-like motion. PMID- 10810157 TI - Molecular determinants of xylose isomerase thermal stability and activity: analysis of thermozymes by site-directed mutagenesis. AB - Xylose isomerases (XIs) from Thermoanaerobacterium thermosulfurigenes (TTXI) and Thermotoga neapolitana (TNXI) are 70.4% identical in their amino acid sequences and have a nearly superimposable crystal structure. Nonetheless, TNXI is much more thermostable than TTXI. Except for a few additional prolines and fewer Asn and Gln residues in TNXI, no other obvious differences in the enzyme structures can explain the differences in their stabilities. TNXI has two additional prolines in the Phe59 loop (Pro58 and Pro62). Mutations Gln58Pro, Ala62Pro and Gln58Pro/Ala62Pro in TTXI and their reverse counterpart mutations in TNXI were constructed by site-directed mutagenesis. Surprisingly, only the Gln58Pro mutation stabilized TTXI. The Ala62Pro and Gln58Pro/Ala62Pro mutations both dramatically destabilized TTXI. Analysis of the three-dimensional (3D) structures of TTXI and its Ala62Pro mutant derivative showed a close van der Waal's contact between Pro62-C(delta) and atom Lys61-C(beta) (2.92 A) thus destabilizing TTXI. All the reverse counterpart mutations destabilized TNXI thus confirming that these two prolines play important roles in TNXI's thermostability. TTXI's active site has been previously engineered to improve its catalytic efficiency toward glucose and increase its thermostability. The same mutations were introduced into TNXI, and similar trends were observed, but to different extents. Val185Thr mutation in TNXI is the most efficient mutant derivative with a 3.1-fold increase in its catalytic efficiency toward glucose. With a maximal activity at 97 degrees C of 45.4 U/mg on glucose, this TNXI mutant derivative is the most active type II XI ever reported. This 'true' glucose isomerase engineered from a native xylose isomerase has now comparable kinetic properties on glucose and xylose. PMID- 10810158 TI - Structure-function analysis of alpha-helix H4 using PSE-4 as a model enzyme representative of class A beta-lactamases. AB - We extracted maximum information for structure-function analysis of the PSE-4 class A beta-lactamase by random replacement mutagenesis of three contiguous codons in the H4 alpha-helix at amino acid positions Ala125, Thr126, Met127, Thr128 and Thr129. These positions were predicted to interact with suicide mechanism-based inhibitors when examining the PSE-4 three-dimensional model. Structure-function studies on positions 125-129 indicated that in PSE-4 these amino acids have a role distinct from those in TEM-1, in tolerating substitutions at Ala125 and being invariant at Met127. The importance of Met127 was suspected to be implicated in a structural role in maintaining the integrity of the H4 alpha-helix structure together, thus maintaining the important Ser130-Asp131 Asn132 motif positioned towards the active site. At the structural level, the H4 region was analyzed using energy minimization of the H4 regions of the PSE-4 YAM mutant and compared with wild-type PSE-4. The Tyr 125 of the mutant YAM formed an edge to face pi-pi interaction with Phe 124 which also interacts with the Trp 210 with the same interactions. Antibiotic susceptibilities showed that amino acid changes in the the H4 alpha-helix region of PSE-4 are particularly sensitive to mechanism based-inhibitors. However, kinetic analysis of PSE-4 showed that the two suicide inhibitors belonging to the penicillanic acid sulfone class, sulbactam and tazobactam, were less affected by changes in the H4 alpha-helix region than clavulanic acid, an inhibitor of the oxypenam class. The analysis of H4 alpha-helix in PSE-4 suggests its importance in interactions with the three clinically useful inhibitors and in general to all class A enzymes. PMID- 10810159 TI - On the possibilities and limitations of rational protein design to expand the specificity of restriction enzymes: a case study employing EcoRV as the target. AB - The restriction endonuclease EcoRV has been characterized in structural and functional terms in great detail. Based on this detailed information we employed a structure-guided approach to engineer variants of EcoRV that should be able to discriminate between differently flanked EcoRV recognition sites. In crystal structures of EcoRV complexed with d(CGGGATATCCC)(2) and d(AAAGATATCTT)(2), Lys104 and Ala181 closely approach the two base pairs flanking the GATATC recognition site and thus were proposed to be a reasonable starting point for the rational extension of site specificity in EcoRV [Horton,N.C. and Perona,J.J. (1998) J. Biol. Chem., 273, 21721-21729]. To test this proposal, several single (K104R, A181E, A181K) and double mutants of EcoRV (K104R/A181E, K104R/A181K) were generated. A detailed characterization of all variants examined shows that only the substitution of Ala181 by Glu leads to a considerably altered selectivity with both oligodeoxynucleotide and macromolecular DNA substrates, but not the predicted one, as these variants prefer cleavage of a TA flanked site over all other sites, under all conditions tested. The substitution of Lys104 by Arg, in contrast, which appeared to be very promising on the basis of the crystallographic analysis, does not lead to variants which differ very much from the EcoRV wild-type enzyme with respect to the flanking sequence preferences. The K104R/A181E and K104R/A181K double mutants show nearly the same preferences as the A181E and A181K single mutants. We conclude that even for the very well characterized restriction enzyme EcoRV, properties that determine specificity and selectivity are difficult to model on the basis of the available structural information. PMID- 10810160 TI - Mutational analyses of restriction endonuclease-HindIII mutant E86K with higher activity and altered specificity. AB - We have performed mutational analyses of restriction endonuclease HindIII in order to identify the amino acid residues responsible for enzyme activity. Four of the seven HindIII mutants, which had His-tag sequences at the N-termini, were expressed in Escherichia coli, and purified to homogeneity. The His-tag sequence did not affect enzyme activity, whereas it hindered binding of the DNA probe in gel retardation assays. A mutant E86K in which Lys was substituted for Glu at residue 86 exhibited high endonuclease activity. Gel retardation assays showed high affinity of this mutant to the DNA probe. Surprisingly, in the presence of a transition metal, Mo(2+) or Mn(2+), the E86K mutant cleaved substrate DNA at a site other than HindIII. Substitution of Glu for Val at residue 106 (V106E), and Asn for Lys at residue 125 (K125N) resulted in a decrease in both endonucleolytic and DNA binding activities of the enzyme. Furthermore, substitution of Leu for Asp at residue 108 (D108L) abolished both HindIII endonuclease and DNA binding activities. CD spectra of the wild type and the two mutants, E86K and D108L, were similar to each other, suggesting that there was little change in conformation as a result of the mutations. These results account for the notion that Asp108 could be directly involved in HindIII catalytic function, and that the substitution at residue 86 may bring about new interactions between DNA and cations. PMID- 10810161 TI - Mutation of recombinant catalytic subunit alpha of the protein kinase CK2 that affects catalytic efficiency and specificity. AB - In order to understand better the structural and functional relations between protein kinase CK2 catalytic subunit, the triphosphate moiety of ATP, the catalytic metal and the peptidic substrate, we built a structural model of Yarrowia lipolytica protein kinase CK2 catalytic subunit using the recently solved three-dimensional structure of the maize enzyme and the structure of cAMP dependent protein kinase peptidic inhibitor (1CDK) as templates. The overall structure of the catalytic subunit is close to the structure solved by Niefind et al. It comprises two lobes, which move relative to each other. The peptide used as substrate is tightly bound to the enzyme, at specific locations. Molecular dynamic calculations in combination with the study of the structural model led us to identify amino acid residues close to the triphosphate moiety of ATP and a residue sufficiently far from the peptide that could be mutated so as to modify the specificity of the enzyme. Site-directed mutagenesis was used to replace by charged residues both glycine-48, a residue located within the glycine-rich loop, involved in binding of ATP phosphate moiety, and glycine-177, a residue close to the active site. Kinetic properties of purified wild-type and mutated subunits were studied with respect to ATP, MgCl(2) and protein kinase CK2 specific peptide substrates. The catalytic efficiency of the G48D mutant increased by factors of 4 for ATP and 17.5 for the RRRADDSDDDDD peptide. The mutant G48K had a low activity with ATP and no detectable activity with peptide substrates and was also inhibited by magnesium. An increased velocity of ADP release by G48D and the building of an electrostatic barrier between ATP and the peptidic substrate in G48K could explain these results. The kinetic properties of the mutant G177K with ATP were not affected, but the catalytic efficiency for the RRRADDSDDDDD substrate increased sixfold. Lysine 177 could interact with the lysine-rich cluster involved in the specificity of protein kinase CK2 towards acidic substrate, thereby increasing its activity. PMID- 10810162 TI - Effect of extra N-terminal residues on the stability and folding of human lysozyme expressed in Pichia pastoris. AB - A human lysozyme expression system by Pichia pastoris was constructed with the expression vector of pPIC9, which contains the alpha-factor signal peptide known for high secretion efficiency. P. pastoris expressed the human lysozyme at about 300 mg/l broth, but four extra residues (Glu(-4)-Ala(-3)-Glu(-2)-Ala(-1)-) were added at the N-terminus of the expressed protein (EAEA-lysozyme). To determine the effect of the four extra residues on the stability, structures and folding of the protein, calorimetry, X-ray crystal analysis and GuHCl denaturation experiments were performed. The calorimetric studies showed that the EAEA lysozyme was destabilized by 9.6 kJ/mol at pH 2.7 compared with the wild-type protein, mainly caused by the substantial decrease in the enthalpy change (DeltaH). On the basis of structural information on the EAEA-lysozyme, thermodynamic analyses show that (1) the addition of the four residues slightly affected the conformation in other parts far from the N-terminus, (2) the large decrease in the enthalpy change due to the conformational changes would be almost compensated by the decrease in the entropy change and (3) the decrease in the Gibbs energy change between the EAEA and wild-type human lysozymes could be explained by the summation of each Gibbs energy change contributing to the stabilizing factors concerning the extra residues. PMID- 10810163 TI - Toast to the Turn of the Century. PMID- 10810164 TI - Croatian Medical Journal at the turn of the millennium. AB - The turn of the millennium coincided with the inclusion of the Croatian Medical Journal into the bibliographic databases MEDLINE (1998), and Current Contents/Clinical Medicine (1999), which greatly increased the number of submitted manuscripts. The increased pressure on the editorial office prompted us to modify the editorial procedure and sharpen our acceptance criteria. At the same time, we extended our author-friendly policy to all for global medicine and (2) medicine in translational and emerging countries. The Editorial Board and the Advisory Board were critical in developing and improving the Journal and setting the highest standards in all aspects of publication, especially in manuscript selection by high-quality peer review. In this editorial, we finally meet the members of the two Boards in person, or rather, in photographs and short biographies. PMID- 10810165 TI - Joining together to combat poverty. AB - The International Poverty and Health Network (IPHN) was created in December 1997 following a series of conferences organized by the World Health Organization, with the aim of integrating health into plans to eradicate poverty. Around 1.3 billion people live on less than US$1 per day. Of the 4.4 billion people in developing countries nearly 60% lack access to sanitation, 30% do not have clean water, 20% have no health care, and 20% do not have enough dietary energy and protein. Even among rich nations there are gross socioeconomic inequalities. Many children are robbed of their physical and mental potential through poverty. Expressed in constant 1963 US dollars, an average Croatian family needed the annual income of US$894 to meet the poverty line in 1960 and US$9,027 in 1995. Accordingly, 9-25% of Croatian households were below the poverty line between 1960 and 1995. The increase in the poverty rate after 1991 was compounded by the war that destroyed almost a third of industrial capacity and infrastructure. Dissipation of the communist economy and inadequate privatization have contributed to the increase in unemployment rate, corruption, and other social ills. IPHN invited Croatian Medical Journal to publish this editorial to help push the issue of poverty up political and medical agendas on a global level. We argue that a factor contributing to the failure of most large-scale programs against poverty to date is the excessive emphasis on material and infrastructure assistance at the expense of spiritual, moral, and intellectual development. PMID- 10810166 TI - Application of forensic DNA testing in the legal system. AB - DNA technology has taken an irreplaceable position in the field of the forensic sciences. Since 1985, when Peter Gill and Alex Jeffreys first applied DNA technology to forensic problems, to the present, more than 50,000 cases worldwide have been solved through the use of DNA based technology. Although the development of DNA typing in forensic science has been extremely rapid, today we are witnessing a new era of DNA technology including automation and miniaturization. In forensic science, DNA analysis has become "the new form of scientific evidence" and has come under public scrutiny and the demand to show competence. More and more courts admit the DNA based evidence. We believe that in the near future this technology will be generally accepted in the legal system. There are two main applications of DNA analysis in forensic medicine: criminal investigation and paternity testing. In this article we present background information on DNA, human genetics, and the application of DNA analysis to legal problems, as well as the commonly applied respective mathematics. PMID- 10810167 TI - Detection and genotyping of Borrelia burgdorferi sensu lato by polymerase chain reaction. AB - AIM: To isolate and genotype Borrelia burgdorferi genospecies in serum samples of Croatian patients with erythema migrans. METHODS: DNA isolates from sera of patients with erythema migrans were analyzed by nested polymerase chain reaction (PCR), amplifying a segment of flagellin gene with primers encompassing the conserved region of the gene. To screen PCR products for heterogeneity, we performed single-stranded conformation polymorphism (SSCP) analysis. The samples showing differences in SSCP patterns were sequenced, and the sequence compared in the GeneBank for sequence homology with known Borrelia burgdorferi genospecies. We also constructed phylogenetic tree of all known borrelial sequences. RESULTS: The nested PCR method using specially designed flagellin gene primers, achieved the sensitivity of 10 genome copies (0.01 pg of purified Borrelia burgdorferi DNA from culture) by dilution analysis. The assay specificity was confirmed by amplification of a part of the flagellin gene from different bacterial species. The primer pairs successfully amplified only Borrelia burgdorferi flagellin gene. The genome of Borrelia burgdorferi sensu lato was detected in the sera of all 10 tested patients with erythema migrans. Sequence data and phylogenetic analysis confirmed that all amplified samples belonged to Borrelia afzelii genospecies. CONCLUSION: Phylogenetic tree analysis placed the borrelial isolates together with Borrelia afzelii sequences into a single group. This finding was additionally supported by sequence homology analysis, which produced a homology score of 99%. In patients with erythema migrans who come from the northwest Croatia, an endemic area for Lyme borreliosis, Borrelia afzelii was the cause of skin manifestations of Lyme borreliosis. PMID- 10810168 TI - Sensitization to non-pyroglyphid mites in urban population of Croatia. AB - AIM: Determination of prevalence of sensitization to Dermatophagoides pteronyssinus, Dermatophagoides farinae (pyroglyphid mites), Lepidoglyphus destructor, and Tyrophagus putrescentiae (non-pyroglyphid mites) in urban continental Croatia. METHODS: Specific IgE (sIgE) and skin prick test (SPT) for D. pteronyssinus, D. farinae, L. destructor, and T. putrescentiae were performed in 67 men. SPT was performed with Epipharm-ALK allergens. A urtica (D+d)/2 of 3 mm or greater was considered a positive skin reaction. SIgE was measured by immunoCAP technology (UNI CAP). Values <0.35 kUA/L were considered nonspecific and >0.35 kUA/L increased. RESULTS: The prevalence of subjects with positive SPT was 35.8% for T. putrescentiae, 26.8% for L. destructor (26.8%), and 22. 4% for D. pteronyssinus and D. farinae. The prevalence of increased sIgE for D. pteronyssinus and D. farinae was 23.9%, followed by T. putrescentiae (22.4%), and L. destructor (14.9%). Among subjects with increased IgE to D. pteronyssinus there were also increased IgE for D. farinae in 94%, for T. putrescentiae in 75%, and for L. destructor in 50%. Mean urtica diameter differed only for L. destructor between nonspecific and increased sIgE values in subjects with positive SPT (3.7+/-0.7 mm vs. 6.2+/-2.8 mm; p = 0.004). Sensitization to storage mites was accompanied by respiratory symptoms in similar proportions as that to house dust mites (51.8% and 54.5%, respectively; p = 0.8527). CONCLUSION: Sensitization to non-pyroglyphid mites is present in the general urban population of Croatia in similar proportions as to pyroglyphid mites, with the same range of respiratory symptoms. Testing with storage mites should be considered routine allergological diagnostic procedure. PMID- 10810169 TI - Radiographic osteoarthritis in the elderly population of Zagreb: distribution, correlates, and the pattern of joint involvement. AB - AIM: To determine the prevalence of radiographic osteoarthritis on five joint groups in an urban population sample of 306 women and 304 men over the age of 45 and to assess the influence of some risk factors on osteoarthritis. The validity of concept of generalized osteoarthritis was also examined by analyzing the association of osteoarthritis on different joint sites. METHODS: Radiographs of both hands, both knees, and the right hip were taken. Osteoarthritic changes on distal interphalangeal, proximal interphalangeal, first carpometacarpal joints, knees, and hip were graded according to the Kellgren-Lawrence scale. The association among osteoarthritis on different joint sites was analyzed using logistic regression. Subjects were tested for age, duration of postmenopause, anthropometric measures, blood pressure, and smoking as risk factors for osteoarthritis. RESULTS: Hip was the most frequent site of osteoarthritis in men (27.3%), whereas distal interphalangeal joints predominated in women (43.5%). Polyarticular osteoarthritis (+/-3 joints) was present in 10.8% women and 5.9% men. There was a significant influence of age on single joint osteoarthritis, but not on multiple joint involvement. Obesity was significantly correlated with knee osteoarthritis in women and with osteoarthritis on distal interphalangeal joints in men. CONCLUSIONS. In our population sample, the prevalence of knee osteoarthritis was lower and the prevalence of hip osteoarthritis higher than reported for most of other populations. The tendency towards polyarticular osteoarthritis that is more common than would be expected by age, suggests a subset of generalized osteoarthritis. PMID- 10810170 TI - Oral cancer in Jordan: a retrospective study of 118 patients. AB - AIM: To evaluate clinical features, diagnosis, and treatment of oral cancer in Jordan. METHOD: We retrospectively analyzed the medical records 118 consecutive patients treated for oral cancer from 1989 until 1998. RESULTS: The age of the patients ranged from 35 to 90 years (median 62.5 years); three quarters were men. Ninety patients (76%) were smokers and 6 (5%) drank alcohol. The floor of the mouth was the most common site for oral cancer, followed by the tongue. The male/female ratio was 3.1:1. The majority of T1 tumors were treated by surgical excision, T2 tumors by surgery or radiotherapy, and T3 and T4 tumors without evidence of nodal disease by radiotherapy. Patients with nodal disease were treated primarily by surgery. In the absence of nodal disease, T1, T2, T3, and T4 tumors had 5-year survival rates of 95%, 95%, 81%, and 25%, respectively, whereas the patients with nodal disease had a poorer prognosis, with survival rates of 37%, 29%, 12% and 0% for T1, T2, T3, and T4, respectively. The 5-year survival rate decreased from 80% to 20% as the stage of disease progressed from I to IV. The overall 5-year survival for all stages of disease was 62/118 (53%). Patients treated with surgery alone (5-year survival rate 62%), and those treated with postoperative radiotherapy (5-year survival rate 81%) did better than patients treated with radiotherapy alone (31%). CONCLUSION: Complete surgical resection combined with radiotherapy results in the best therapeutic outcome. PMID- 10810171 TI - Retrospective analysis of pectoralis major myocutaneous flap surgeries performed under war conditions. AB - AIM: To present our experiences in using a pectoralis major myocutaneous flap in the reconstruction of surgically created defects of the neck and lower part of the head during the war in Bosnia and Herzegovina. METHODS: Retrospective analysis of medical records from 15 patients treated at the ENT Department, Tuzla University Hospital, between January 1992 and December 1996. RESULTS: Ten flaps were prepared during basic operation ("one step reconstruction of defect") and five flaps three weeks after the removal of tumor (postoperative pharyngocutaneous fistula was the reason for secondary flap preparation). The necessary time for flap preparation and its accommodation in the defect was 2 hours. The most frequent complications included seroma of donor site (6/13), fistula (3/13), partial necrosis of the flap (2/13), and total necrosis of the flap (2/13). Three patients died in the postoperative period because of a cardiac arrest but the flaps accepted correctly. CONCLUSION: The pectoralis major myocutaneous flap is a good solution for covering defects of the neck and lower region of the head. Military blockade with extremely difficult conditions and lack of experience were the reason of the higher complication rate than those from literature. PMID- 10810172 TI - Color Doppler flow imaging of ocular tumors. AB - AIM: To analyze the usefulness of color Doppler flow imaging in the differentiation of benign and malign ocular tumors. METHODS: Blood flow in tumor and ocular blood vessels was assessed by color Doppler flow imaging in 20 patients with malignant melanoma of the uvea and 19 patients with cavernous hemangioma. Blood velocity measurements in orbital vessels in these patients were compared with the same measurements in 20 healthy individuals. RESULTS: Blood flow inside the tumor tissue was observed in all patients, except in a single case of uveal melanoma. Internal blood flow of intraorbital hemangiomas was slower and their resistance index lower than that of melanomas. Patients with a tumor did not differ from the controls in blood flow velocities in the ophthalmic artery, central retinal artery, and posterior ciliary arteries. Only the patients with uveal melanoma had higher maximal blood flow in central retinal artery and posterior ciliary arteries compared to other patients. CONCLUSION: Color Doppler flow imaging of blood flow can be a useful method to assess the vascularization of ophthalmological tumors, and to differentiate uveal melanoma from orbital hemangioma. PMID- 10810173 TI - Platelet monoamine oxidase activity, ego strength, and neuroticism in soldiers with combat-related current posttraumatic stress disorder. AB - AIM: To assess possible differences in platelet monoamino oxidase-B (MAO-B) activity, ego strength, and neuroticism in combat-experienced soldiers with or without current posttraumatic stress disorder (PTSD). METHOD: The soldiers with current PTSD (N=36) and a control group of 34 healthy soldiers were matched in combat experience, time passed between combat experience and the study, demographic variables (age, marital status, education), and smoking status. Platelet MAO-B was assayed fluorometrically, ego strength was measured by the Croatian version of the Ego Identity Scale, and neuroticism by the N-scale from Eysenck's EPQ-R questionnaire. RESULTS: Soldiers with combat-related current PTSD had lower platelet MAO-B activity than the control group (9.1+/-3.9 vs. 11.9+/ 4.0; p<0.05), as well as lower ego-strength (86.3+/-8.3 vs. 108.6+/-13.4; p<0.05) and higher neuroticism (23.5+/-13.2 vs. 5. 9+/-4.7; p<0.05). There was no association of ego strength or neuroticism with platelet MAO-B activity. CONCLUSION: Ego identity strength and emotional stability are associated with successful coping with combat trauma. The involvement of platelet MAO-B activity in biological basis of ego strength and neuroticism could not be demonstrated. PMID- 10810174 TI - Public health reforms and development of health service in the Dakovo sub district and administrative county in the period 1850-1899. AB - In the long-lasting struggle for national identity and modernization of Croatia, the Parliament of the Kingdom of Croatia and Slavonia has passed many laws and regulations from 1874 on, affecting thus the health care and the development of the public health system. The aim of those laws was to establish and achieve the same level of public health care that had already been instituted in some other countries of the Austro-Hungarian Empire. In order to clarify the consequences of these reforms for the development of the health care system on the county and district levels of Slavonia, we collected data on the town of Dakovo as a market center, home of the diocese, and seat of the sub-district and administrative county. The data were divided into several categories in order to examine (1) the reorganization of health care in the Kingdom of Croatia and Slavonia from 1848 to 1894; (2) the development of health care in the Dakovo sub-district and Dakovo administrative county; (3) number, structure, and distribution of medical practitioners from 1807 to 1899; (4) hospitals from 1859 to 1900; and (5) selected indicators of health and living condition and health needs of the county inhabitants in the period 1850-1900. The analysis of historical material showed that new regulations of health care initiated the process of "medicalization" that was understood as a part of European modernization in the field of state medicine and health care administration. It brought more accurate knowledge of the main causes of illnesses, deaths and disabilities but did not significantly improve health and health conditions in the Dakovo County at the entrance of the 20th century. PMID- 10810175 TI - Pheochromocytoma with negative urinalysis in pregnancy. AB - We present a rare case of adrenal pheochromocytoma in pregnancy, with serial 24-h urine specimen collections showing normal concentrations of catecholamine metabolites. The diagnosis was based on clinical presentation, abdominal ultrasound, and magnetic resonance imaging, and was confirmed on post-operative pathohistological examination. Clinical suspicion of pheochromocytoma in pregnancy should be sufficient to implement adequate therapeutic measures, regardless of urine catecholamine concentrations. PMID- 10810176 TI - Ankyrin: structure, properties, and functions. AB - Recent data on characteristics of the structure, functions, and main properties of ankyrins (proteins that are linkers between the spectrin-based cytoskeleton and integral membrane proteins) are summarized. The interactions of ankyrins with band-3 protein, P-type ATPases, ion channels, receptors, and protein kinase C are considered. The structure of ankyrin repeats that are often contained in other proteins (which are not classified with the ankyrin family) and ensure protein protein interactions as well as interactions between proteins and nucleic acids is described in details. The mechanisms of regulation of the ability of ankyrins to interact with other proteins (alternative splicing and post-translational modification, including phosphorylation) are also considered. PMID- 10810177 TI - Nitric oxide and NO-synthases in mammals in different functional states. AB - This review deals with the physical and chemical properties of nitric oxide as well as with the mechanisms and enzymes synthesizing this compound in animals including humans. The cytotoxic, vasodilatory, neuromediatory, and other properties of NO are analyzed. Polyfunctionality of NO in the norm and in pathologies of different genesis is shown. It is suggested that the analysis of the mechanisms of cyclic conversion of nitric oxide and the elucidation of the role of all NO metabolism products in living organisms would allow us to approach a more profound understanding of the NO problem in biology and medicine. It is reasonable to think the knowledge obtained in the course of the studies will permit an elaboration of the strategy and tactics of medical treatment of many diseases occurring on the background of disturbance in the mechanisms of formation and utilization of this compound. PMID- 10810178 TI - Bioactive amide of prostaglandin E1 and ethanolamine plasmalogen analog of platelet-activating factor inhibits several pathways of human platelet aggregation. AB - The influence of an amide of prostaglandin E1 and ethanolamine plasmalogen platelet-activating factor analog 1-O-alk-1;-enyl-2-acetyl-sn-glycero-3-phospho (N-11alpha, 15alpha-dioxy-9-keto-13-prostenoyl)ethanolamine (PGE1-PPAF) on platelet-activating factor (PAF)-, ADP-, and thrombin-induced human platelet aggregation has been studied. It was found that PGE1-PPAF inhibits the PAF-, ADP , and thrombin-induced platelet aggregation in platelet-rich plasma. 1-O-alk-1; enyl-2-acetyl-sn-glycero-3-phosphoethanolamine inhibited PAF-induced aggregation up to 50% but had no influence on platelet aggregation induced by ADP or thrombin. The ethanolamine plasmalogen analog of PAF 1-O-alk-1;-enyl-2-acetyl-sn glycero-3-phospho-(N-palmitoyl)ethanolami ne, having a palmitoyl residue instead of PGE1, did not inhibit platelet aggregation induced by PAF, ADP, or thrombin. We propose that inhibition of human platelet aggregation by PGE1-PPAF is mediated by its action on platelet PAF-receptors and the adenylate cyclase system. PMID- 10810179 TI - Role of acetylcholine in regulation of interaction between axon and Schwann cell during rhythmic excitation of nerve fibers. AB - Axon excitation increases the number of acetylcholine receptors (ACR) of the Schwann cell (SC) depending on the frequency of rhythmic excitation (RE) and on intercellular concentrations of K+, Ca2+, and acetylcholine. During RE, activity of axonal acetylcholine esterase is decreased, thus providing for high intercellular acetylcholine concentration. Increased intercellular concentration of acetylcholine activates phosphoinositide-specific phospholipase C (PIPLC) of the myelin nerve fiber. During RE, K+ depolarization and acetylcholine exocytosis can activate Ca2+ entry via Ca2+ channels, thus inducing SC ACR phosphorylation mediated by PIPLC stimulation. PMID- 10810180 TI - Phenylalanyl-tRNA synthetase interacts with DNA: studies on activity using deoxyribooligonucleotides. AB - Phenylalanyl-tRNA synthetase from Thermus thermophilus complexes with short (up to 30 nucleotide length) single-stranded DNA fragments more efficiently than with double-stranded fragments. The complexing between DNA and the protein significantly increases with deoxyribooligonucleotide longer than 20 nucleotides. Using affinity labeling, the binding site of DNA was located near the interface of the alpha- and beta-subunits. The binding sites of DNA and tRNAPhe do not overlap. Phenylalanyl-tRNA synthetase from E. coli also binds DNA. PMID- 10810181 TI - Changes in superoxide production rate and in superoxide dismutase and glutathione peroxidase activities in subcellular organelles in mouse liver under exposure to low doses of low-intensity radiation. AB - The functioning of the antioxidant system in mouse liver at increased stationary concentration of active oxygen species induced by whole-body chronic exposure of mice to gamma-irradiation (137Cs, 0.6 cGy/day, 9 days) was studied. Synchronous changes (growth with an extreme) in activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) are found that may be considered as evidence in favor of maintenance of regulatory links in the antioxidant system of liver. The superoxide production rate in microsomes and nuclei also changed with an extreme with the rise in accumulated radiation dose. In microsomes the superoxide production rate reached a maximum at lower doses than the activity of Cu,Zn-SOD did. In nuclei the increase in superoxide production rate was not compensated by the rise in Cu,Zn-SOD activity within the studied dose range. The findings indicate some imbalance between production and consumption of superoxide radicals in microsomes and nuclei; in mitochondria these processes are balanced, leading to more resistance to low-dose irradiation. PMID- 10810182 TI - Uterine myometrial ferritin and blood plasma transferrin as natural modulators of Ca-calmodulin-independent cyclic nucleotide phosphodiesterase from cow uterine myometrium. AB - The iron storage protein ferritin was purified to electrophoretic homogeneity from cow uterine myometrium. Its Mr did not exceed 440, 000 and H-chains predominated in the subunit composition; the iron saturation was 43 iron ions per protein molecule. The uterine myometrial ferritin was a potent natural modulator of Ca-calmodulin-independent phosphodiesterase (Ca-CM-independent PDE, EC 3.1.4.17) isolated from the same tissue. Addition of iron-poor ferritin from uterine myometrium and iron-reach liver ferritin caused three- and two-fold inhibition of the enzyme activity, respectively. The iron transport protein transferrin in iron-saturated and iron-depleted forms can also inhibit Ca-CM independent PDE activity by two-fold. In both cases, the degree of saturation with iron was not crucial for the inhibitory effects of these proteins on the enzyme activity. These data suggest that iron homeostasis proteins can modulate the cyclic nucleotide level in non-nervous tissue via interaction with enzymes involved in cyclic nucleotide hydrolysis. PMID- 10810183 TI - Effect of methanol on the activity and conformation of acid phosphatase from the prawn Penaeus penicillatus. AB - Prawn (Penaeus penicillatus) acid phosphatase (EC 3.1.3.2) catalyzes the nonspecific hydrolysis of phosphate monoesters. The effects of some pollutants in sea water on the enzyme activity results in the loss of the biological function of the enzyme, which leads to disruption of phosphate metabolism in cells. This paper analyzes the effects of methanol on the activity and conformation of prawn acid phosphatase. The results show that low concentrations of methanol can lead to reversible inactivation. Inhibition of the enzyme by methanol is classified as non-competitive inhibition, and the inhibition constant (Ki) is 8.5%. Conformational changes of the enzyme molecule in methanol solutions of different concentrations were measured using fluorescence emission, differential UV absorption, and circular dichroism spectra. Increased methanol concentrations caused the fluorescence emission intensity of the enzyme to increase. The ultraviolet difference spectra of the enzyme denatured with methanol had two negative peaks, at 222 and 270 nm, and a positive peak at 236 nm. The changes in the fluorescence and ultraviolet difference spectra reflected the changes of the microenvironments of tryptophan and tyrosine residues of the enzyme. The CD spectrum changes of the enzyme show that the secondary structure of the enzyme also changed some. These results suggest that methanol is a non-competitive inhibitor and the conformational integrity of the enzyme is essential for its activity. PMID- 10810184 TI - Benzodifuroxan as an NO-dependent activator of soluble guanylate cyclase and a novel highly effective inhibitor of platelet aggregation. AB - The ability of benzodifuroxan (BDF) to activate human platelet guanylate cyclase was investigated. The maximal stimulatory effect (1160 +/- 86%) was observed at 0.01 mM concentration. Sodium nitroprusside produced the same stimulatory effect (1220 +/- 100%) but at a higher concentration (0.1 mM). 1-H-[1,2,4,] Oxadiazolo[4, 3-alpha]quinoxalin-1-one (ODQ), an inhibitor of NO-dependent guanylate cyclase activation, attenuated the stimulatory effect of BDF (0.01 mM) by 75% and that of sodium nitroprusside (0.1 mM) by 80%. Increasing dithiothreitol concentration in the sample from 2. 10-6 to 2.10-4 M increased the stimulatory effect of BDF 2.7-fold. The possible involvement of sulfhydryl groups of low-molecular-weight thiols and guanylate cyclase in thiol-dependent activation of the enzyme is discussed. We have also found that BDF is a highly effective inhibitor of ADP-induced human platelet aggregation with IC50 of 6.10-8 M. The effect of sodium nitroprusside was much weaker (IC50, 5.10-5 M). PMID- 10810185 TI - Participation of glyceraldehyde-3-phosphate dehydrogenase in the regulation of 2,3-diphosphoglycerate level in erythrocytes. AB - Data are presented concerning the possible participation of glyceraldehyde-3 phosphate dehydrogenase (GAPDH) in regulation of the glycolytic pathway and the level of 2,3-diphosphoglycerate in erythrocytes. Experimental support has been obtained for the hypothesis according to which a mild oxidation of GAPDH must result in acceleration of glycolysis and in decrease in the level of 2, 3 diphosphoglycerate due to the acyl phosphatase activity of the mildly oxidized enzyme. Incubation of erythrocytes in the presence of 1 mM hydrogen peroxide decreases 2,3-diphosphoglycerate concentration and causes accumulation of 3 phosphoglycerate. It is assumed that the acceleration of glycolysis in the presence of oxidative agents described previously by a number of authors could be attributed to the acyl phosphatase activity of GAPDH. A pH-dependent complexing of GAPDH and 3-phosphoglycerate kinase or 2, 3-diphosphoglycerate mutase is found to determine the fate of 1,3-diphosphoglycerate that serves as a substrate for the synthesis of 2,3-diphosphoglycerate as well as for the 3-phosphoglycerate kinase reaction in glycolysis. A withdrawal of the two-enzyme complexes from the erythrocyte lysates using Sepharose-bound anti-GAPDH antibodies prevents the pH dependent accumulation of the metabolites. The role of GAPDH in the regulation of glycolysis and the level of 2,3-diphosphoglycerate in erythrocytes is discussed. PMID- 10810186 TI - Two new site-specific endonucleases from Staphylococcus species strain D5. AB - Staphylococcus species strain D5 containing two site-specific endonucleases, SspD5 I and SspD5 II, was found during screening of a bacterial strain collection from soil. These endonucleases were purified to functional homogeneity by sequential chromatography. Site-specific endonuclease SspD5 I recognizes sequence 5;-GGTGA(8N/8N) downward arrow-3; on DNA. Unlike Hph I, it cleaves DNA at a distance of 8 nucleotides from the recognized sequence on both chains producing blunt-end DNA fragments, while endonuclease Hph I cleaves DNA forming mononucleotide 3;-OH protruding ends. Thus, endonuclease SspD5 I is a new type II site-specific endonuclease and a neoschizomer of endonuclease Hph I. The advantage of this new endonuclease is that the blunt-end DNA products of this enzyme can be inserted without additional treatment into vector DNAs cleaved with endonucleases yielding DNA blunt-ends. Endonuclease SspD5 II recognizes site 5' ATGCA T-3' and thus is an isoschizomer of endonuclease Nsi I. The molecular mass of SspD5 I is about 35 kD and that of SspD5 II is 40 kD. The enzymes exhibit maximal activity at 37 degrees C. The optimal buffer for the reaction is HRB (10 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 100 mM NaCl, and 1 mM dithiothreitol). PMID- 10810187 TI - Generation of transmembrane electrical potential during NADH oxidation via the external pathway and the fatty acid uncoupling effect after transient opening of the Ca2+-dependent cyclosporin A-sensitive pore in liver mitochondria. AB - The effects of transient pore opening on generation of the transmembrane gradient of electrical potential across the inner mitochondrial membrane (DeltaPsi) induced by NADH oxidation through the external pathway as well as on the uncoupling effect of fatty acids were studied. The pore opening was monitored by changes in the DeltaPsi value. The cycle of pore opening/closing was found to have only an insignificant effect on the sensitivity of DeltaPsi to fatty acid uncoupling. Once this cycle is over, NADH oxidation in the presence of exogenous cytochrome c results in generation of DeltaPsi. In the absence of cytochrome c, the generation of DeltaPsi induced by oxidation of exogenous NADH is observed if the incubation medium pH has been decreased from 7.4 to 7.0. The generation of DeltaPsi was inhibited by cyclosporin A. In isotonic salt medium containing 125 mM KCl, the maximum level of DeltaPsi generated by exogenous NADH after the cycle of pore opening/closing was significantly lower than the maximum level of DeltaPsi generated in hypotonic incubation medium. The data obtained in this work suggest that the cycle of pore opening/closing has little if any effect on the energy coupling in liver mitochondria, whereas the external pathway of NADH oxidation activated by this cycle may support the energy-dependent functions of liver mitochondria. PMID- 10810188 TI - Interaction of porin from Yersinia pseudotuberculosis with lipopolysaccharides. Effect of ionic strength, pH, and divalent cations on the binding parameters. AB - Interaction of the pore-forming protein (porin) from Yersinia pseudotuberculosis with S- and R-forms of the endogenous lipopolysaccharide (LPS) was studied at various ionic strengths (20-600 mM NaCl), concentrations of divalent cations (5 100 mM CaCl2, MgCl2), and pH values from 3.0 to 9.0. The interaction of the R-LPS with porin has been shown in all experimental conditions to be in consensus with the model suggesting binding at independent sites of two types. S-LPS binds to interacting sites of relatively high affinity and to independent sites of low affinity at all pH values examined and at low NaCl concentration. The cooperative interaction of the S-LPS and porin is not observed at high ionic strength and in divalent cation-free medium. The number of binding sites of porin and association constants (Ka) for both LPS forms decrease significantly on increasing the solution ionic strength. The Ka values for the R- and S-LPS change oppositely on changing the pH: the Ka value for the R-LPS is maximal (Ka = 6.7 x 10(5) M-1), but that for S-LPS is minimal (Ka = 0.4 x 10(5) M(-1) at pH 5.0-5.5. The number of high-affinity and low-affinity binding sites for both LPS forms is maximal at pH 5.0-5.5. In this case, the numbers of high- and low-affinity sites for R-LPS are 3 and 10, respectively, and those for the S-LPS are 7 and 20, respectively. These data suggest an important role of electrostatic interactions on binding of LPS to porin. The contribution of conformational changes of the ligand and protein and hydrophobic interactions are discussed. PMID- 10810189 TI - Carbohydrate-carbohydrate interaction: zymosan and beta-glucan from Saccharomyces cerevisiae bind mannosylated glycoconjugates. AB - Zymosan from cell wall of the yeast Saccharomyces cerevisiae was found to interact with synthetic and natural glycoconjugates; mannose-rich glycoprotein N chains demonstrated the maximal affinity. The carbohydrate-carbohydrate nature of the interaction has been confirmed by the following data: 1) periodate treatment of the zymosan impairs the binding; 2) neither protease nor alkali treatment of the zymosan weaken the binding; 3) beta-glucan from S. cerevisiae, which is the major zymosan component, interacts with glycoconjugates similarly to zymosan. The binding is reversible, Ca2+-dependent, and cooperative; it can be dose dependently inhibited by saccharides relative to one of the partners of the carbohydrate-carbohydrate interaction. PMID- 10810190 TI - Metabolic control of sodium transport in streptozotocin-induced diabetic rat hearts. AB - Diabetic and control cardiomyocytes encapsulated in agarose beads and superfused with modified medium 199 were studied with 23Na- and 31P-NMR. Baseline intracellular Na+ was higher in diabetic (0.076 +/- 0.01 micromoles/mg protein) than in control (0.04 +/- 0.01 micromoles/mg protein) (p < 0.05). Baseline betaATP and phosphocreatine (PCr) (peak area divided by the peak area of the standard, methylene diphosphonate) were lower in diabetic than in control, e.g., betaATP control, 0.70 +/- 0.07; betaATP diabetic, 0. 49 +/- 0.04 (p < 0.027); PCr control, 1.20 +/- 0.13; PCr diabetic, 0. 83 +/- 0.11 (p < 0.03). This suggests that diabetic cardiomyocytes have depressed bioenergetic function, which may contribute to abnormal Na,K-ATPase function, and thus, an increase in intracellular Na+. In the experiments presented herein, three interventions (2 deoxyglucose, dinitrophenol, or ouabain infusions) were used to determine whether, and the extent to which, energy deficits or abnormalities in Na,K-ATPase function contribute to the increase in intracellular Na+. In diabetic cardiomyocytes, 2-deoxyglucose and ouabain had minimal effect on intracellular Na+, suggesting baseline depression of, or resetting of both glycolytic and Na,K ATPase function, whereas in control both agents caused significant increases in intracellular Na+after 63 min exposure: 2-deoxyglucose control, 32.9 +/- 8.1%; 2 deoxyglucose diabetic, -4.6 +/- 6% (p < 0.05); ouabain control, 50.5 +/- 8.8%; ouabain diabetic, 21.2 +/- 9.2% (p < 0.05). In both animal models, dinitrophenol was associated with large increases in intracellular Na+: control, 119.0 +/- 26.9%; diabetic, 138.2 +/- 12.6%. Except for the dinitrophenol intervention, where betaATP and PCr decreased to levels below 31P-NMR detection, the energetic metabolites were not lowered to levels that would compromise sarcolemmal function (Na,K-ATPase) in either control or diabetic cardiomyocytes. In conclusion, in diabetic cardiomyocytes, even though abnormal glycolytic and Na, K-ATPase function was associated with increases in intracellular Na+, these increases were not directly related to global energy deficit. PMID- 10810191 TI - Structure of a teichoic acid from Nocardioides luteus VKM Ac-1246T cell wall. AB - A teichoic acid from the cell walls of Nocardioides luteus VKM Ac-1246T, a validly described species of the Nocardioides genus, is a 1,5-poly(ribitol phosphate) completely substituted at C-4 by alpha-D-galactopyranosyl residues carrying a 4,6-pyruvate ketal group in R-configuration. The structure of the repeating unit of the polymer is as follows: [figure]. The chain consists of approximately 18 repeating units and six beta-D-galactofuranosyl residues linked in the oligomer by 1,6-glycosidic bonds. The oligomer probably terminates the growing end of the teichoic acid. The structure of the polymer was determined by chemical methods and NMR spectroscopy. This teichoic acid has not been described so far. PMID- 10810192 TI - [Chest imaging in 2000: past, present, and future]. PMID- 10810193 TI - [Standard chest x-ray: from film to all digital!]. AB - Conventional imaging will be totally replaced by digital imaging because of the lower exposure dosage and the clear advantages in terms of image processing, transfer and storage. Reports in the literature have demonstrated that a 2.5 pl/mm spatial resolution is satisfactory to detect a pneumothorax, a parenchymal nodule or a mediastinal mass. For most authors, this resolution is also sufficient to detect an interstitial syndrome. Digitalization by amplification or by particle detectors using wire chambers are not, for the time being, acceptable for interpretation of a simple chest image because of the low spatial resolution. The selenium cylinder is an adapted technology dedicated to chest imaging. Memory display screens can be used to digitalize the entire radiology unit. They are perfectly adapted for bedside imaging. Flat detectors are based on direct or indirect conversion of the X photons into an electrical signal during the clinical evaluation. The early results appear most promising. PMID- 10810194 TI - [Computed tomography in 2000: technique, expected progress, limitations, indications]. AB - Computed tomography (CT) is today a fundamental technique for thoracic imaging, a logical direct extension of the standard chest x-ray. Since the early 90s, volume acquisition CT has greatly improved the performance of the conventional technique. After replacing lung and bronchial tomography, the technique has greatly contributed to the reduction in the number of diagnostic conventional angiographies. Certain decisional algorithms have been totally changed, particularly for the diagnosis of pulmonary embolism. In addition, technological advances offer new perspectives, particularly in the field of morphofunctional imaging. There are still many indications for CT and new applications can be expected. Possible drawbacks related to radiation dose and injection of contrast agents must be kept in mind. PMID- 10810195 TI - [Magnetic resonance angiography of the thorax in 2000]. AB - Over the last five years, magnetic resonance imaging (MRI) has been the scene of explosive developments in many fields. MRI exploration of thoracic vessels, perfusion imaging of the pulmonary parenchyma, cine-MR, and apnea sequences have all seen impressive advances. This improvement has been made possible by progress in data processing and surface coil technology for a better signal-noise ratio. In 2000, indications for thoracic MRI remain limited due to the small number of machines available and the fact that the CT scan can provide superior quality images for the pulmonary parenchyma and for visualizing calcifications. MRI can be a complementary exam for lung cancer work-up in case of suspected invasion of the thoracic cavity or the mediastinum. It is the first intention exam in cases involving the greater vessels, with the exception of emergency situations, the posterior mediastinum, and the heart. We review here the current contribution of MRI in diseases of the chest and discuss perspectives for the future. PMID- 10810196 TI - [Thoracic ultrasound]. AB - Ultrasound exploration of the thorax is a very useful imaging method in children. For adults, it is useful for pleural effusion, study of the diaphragm and to guide percutaneous puncture of targeted pleuro-parietal, mediastinal or peripheral parenchymal sites. Thoracic ultrasound remains underused, particularly as a bedside exploration technique or in emergency or intensive care situations. PMID- 10810197 TI - [Interventional radiology of the thorax]. AB - Interventional radiology of the thorax includes a wide variety of diagnostic and/or therapeutic procedures. Transthoracic needle biopsy is a widely used technique. Generall, computed tomography is used to guide the puncture, less commonly fluoroscopy or in exceptional cases ultrasound. Targets include parenchymatous, pleural and mediastinal lesions. The biopsy specimen is used for cytology, histology, bacteriology, mycology and parasitology studies. Needle biopsy is also the first step in more complex procedures: drainage of thoracic collections, inserting harpoons, thoracic sympatholysis, palliative treatment of aspergillomas. Bronchial arteriography with embolization is an effective emergency symptomatic treatment for severe hemoptysis. Endovascular vaso occlusion procedures can be used for rare arteriovenous pulmonary fistulas. Superior vena cava obstruction can be treated, usually in a palliative procedure, by installing an endovascular endoprosthesis. PMID- 10810198 TI - [Contribution of positron emission tomography for the management of lung cancer]. AB - 18 FDG- PET is an imagining technique based on metabolic criteria rather than morphological criteria. (18) FDG- PET can demonstrate accelerated glycosis in cancer tissue related to increased transporter and glycolytic enzyme activity. Whole body PET is currently under validation in a growing number of indications during diagnostic and therapeutic assessment phases of cancer treatment. In the field of pulmonary oncology, (18) FDG- PET has already demonstrated its performance capacity to: 1) discriminate the malignant nature of a solitary pulmonary nodule, 2) improve sensitivity over CT for mediastinal assessment in small-cell lung cancer, 3) acquire whole body imaging to search for distant metastasis in patients with small-cell lung cancer; PET is particularly useful for evaluation of an adrenal mass, 4) complement CT imaging to better dissociate tumor residue or recurrence from post-therapeutic sequelae in small-cell lung cancer. Information provided by (18) FDG- PET is thus clinically relevant as it allows better dissociation of a benign process from a malignant process and better precision of small-cell lung cancer extension without necessitating systematic invasive exploration. PMID- 10810199 TI - [3D virtual imaging of the upper airways]. AB - The different three dimensional reconstructions of the upper airways that can be obtained with spiral computed tomograpy (CT) are presented here. The parameters indispensable to achieve as real as possible spiral CT images are recalled together with the advantages and disadvantages of the different techniues. Multislice reconstruction (MSR) produces slices in different planes of space with the high contrast of CT slices. They provide information similar to that obtained for the rare indications for thoracic MRI. Thick slice reconstructions with maximum intensity projection (MIP) or minimum intensity projection (minIP) give projection views where the contrast can be modified by selecting the more dense (MIP) or less dense (minIP) voxels. They find their application in the exploration of the upper airways. Surface and volume external 3D reconstructions can be obtained. They give an overall view of the upper airways, similar to a bronchogram. Virtual endoscopy reproduces real endoscopic images but cannot provide information on the aspect of the mucosa or biopsy specimens. It offers possible applications for preparing, guiding and controlling interventional fibroscopy procedures. PMID- 10810200 TI - [Towards the integration of the digital medical image folder within the computerized patient folder: PACS and image networks]. AB - Medical Images are components of the so-called "Medical Imaging Folder". This folder is a subset of the so-called "Medical Folder", part of the "Patient Folder. The G8 promotes the concept of a "Global Information Society for Health. Within this society, the Patient Folder is seen either from a healthcare organization, from a country or from an international point of view. The Global Patient Folder (Healthcare Folder) is composed of the different Patient Folder instances. Presently, Pacs and Telemedicine are no longer concerned only by storage and transmission issues. The medical images have only meaning associated with their context, the patient healthcare status. This context is rich in information provided by various information systems or healthcare professionals. The different data are linked and time dependant. Therefore, the expert community in the field of patient records argues that the approach must be the integration of Medical Images within the patient folder. It appears clearly that the complete deployment of such an "International Healthcare Folder" needs time and will proceed in several steps. Due to the increase of the people's mobility this deployment is inescapable. Infrastructure must be sized up taking into account the Digital Medical Image spreading and its large data volume which necessitates a large bandwith. In this paper, we detail the Medical Image Folder concept and its position within the Patient Folder and the Healthcare Folder. Then we present PACS, networking and Telemedicine concepts as well as the needs in standards. PMID- 10810201 TI - [Medico-legal aspects of thoracic imaging in 2000]. AB - The radiologist's responsibility for the radiographic procedure concerns control of the indication and patient information and consent, as well as the examination procedure itself, including safety measures, interpretation, report delivery, and patient follow-up until referral to a colleague. Details on five specific medico legal points are examined here: 1) patient information and consent, a problem recently brought to the forefront by the recent French Supreme Court decision, 2) lung cancer unrecognized on a radiography, 3) implications of complementary examinations related to the discovery of a suspect lesion, 4) fortuitous discovery of a malignant lesion in a patient undergoing exploration for another reason, 5) questions of a legal and ethical nature raised by the development of telemedicine. PMID- 10810202 TI - The regulation of apoptosis in preantral ovarian follicles. AB - Less than 1% of ovarian follicles ever mature to ovulation. The remainder undergo atretic degeneration via apoptosis during development. Though the regulation of antral and preovulatory survival has been studied for many years, very little is known about the regulation of survival and development of preantral follicles. This review discusses recent findings regarding preantral follicle development with emphasis on the regulation of preantral follicle apoptosis. PMID- 10810204 TI - Granulosa cell apoptosis: conservation of cell signaling in an avian ovarian model system. AB - Ovarian follicle atresia in all vertebrates studied to date is mediated via apoptosis, a process that is often initiated within the granulosa cell layer. While follicle atresia is considered a normal physiological process to insure the greatest chance for ovulation of fertilizable oocytes, abnormally high rates of atresia result in chronic infertility and/or premature termination of fertility (e.g., menopause). Although the vast majority of research to elucidate the molecular ordering of cell signaling during the process of granulosa cell apoptosis has been conducted in mammalian model systems, there is ample evidence to demonstrate that many of the proteins, enzymes and cell-signaling pathways are common to ovarian follicles from avian species. The following review will discuss evidence for the conservation of cellular processes that regulate the fate of granulosa cells from the avian, versus mammalian, ovary during follicle development. PMID- 10810203 TI - Mammalian follicular development and atresia: role of apoptosis. AB - The regulation of follicular development and atresia is a complex process and involves interactions between endocrine factors (gonadotropins) and intraovarian regulators (sex steroids, growth factors and cytokines) in the control of follicular cell fate (i.e. proliferation, differentiation and programmed cell death). Granulosa and theca cells are key players in this fascinating process. As atresia is the fate of most follicles, understanding of how these physiological regulators participate in determining the destiny of the follicle (to degenerate or to ovulate) at cellular and subcellular levels is fundamental. This short review summarizes the role of intraovarian modulators of programmed cell death in the induction of atresia during follicular development. PMID- 10810205 TI - Proteolytic and cellular death mechanisms in ovulatory ovarian rupture. AB - Collagen breakdown and cellular death (apoptosis and inflammatory necrosis) within the apex of preovulatory ovine follicles are hallmarks of impending ovarian rupture. An integrative mechanism is presented whereby gonadotropic stimulation of urokinase-type plasminogen activator secretion by ovarian surface epithelial cells bordering the preovulatory follicle elicits a localized increase in tissue plasmin, which activates latent collagenases and secretion of tumor necrosis factor-alpha (TNF-alpha) from thecal endothelium. TNF-alpha potentiates collagenolysis (via matrix metalloproteinase gene expression) and (at elevated concentrations) mediates epithelial/vascular dissolution. Incidental damage to DNA of ovarian surface epithelial cells circumjacent to the ruptured follicle is a putative etiological factor in ovarian cancer. PMID- 10810206 TI - N-cadherin-mediated cell contact regulates ovarian surface epithelial cell survival. AB - The present study shows that in cells derived from the rat ovarian surface epithelium (i.e. ROSE-179 cells) the adhesion protein, N-cadherin, binds to the receptor for fibroblast growth factor (FGF). This interaction likely accounts for the ability of the N-cadherin antibody to decrease the activation (i.e. tyrosine phosphorylation) of the FGF receptor and induce apoptosis. The loss of N-cadherin mediated cell contact also results in the accumulation of beta-catenin within the nucleus. Since beta-catenin also functions as a transcription factor, nuclear beta-catenin may promote mRNA synthesis that is required for ROSE-179 cells to undergo apoptosis in response to serum/calcium withdrawal. This hypothesis requires further testing and validation. PMID- 10810207 TI - Apoptosis and chemoresistance in human ovarian cancer: is Xiap a determinant? AB - Cisplatin-induced apoptosis in epithelial ovarian cancer cells is in part a consequence of suppressed Xiap expression and upregulation of the Fas/FasL system. Changes in the expression of these 'cell death' and 'cell survival' genes lead to activation of caspase-3, and cleavage of MDM2 and FAK. Failure of cancer cells to maintain a balance in the expression of these genes in favor of apoptotic cell death may be an important factor of chemoresistance. Xiap may be a novel target for gene therapy of human ovarian epithelial cancer and, dependent on P53 status, expression of Xiap antisense alone or in combination with wild type P53 sense may offer a new approach for the treatment of the chemoresistant cancer. PMID- 10810208 TI - Pharmacodynamics and pharmacokinetics of levofloxacin. AB - Principles of antibiotic pharmacodynamics include factors that are important for effective eradication of bacteria as well as the suppression of resistance. For effective eradication of bacteria and a good clinical outcome, a ratio for the area under the plasma concentration-time curve relative to the minimum inhibitory concentration (AUC/MIC) of greater than 100 is needed for gram-negative organisms, and a level of greater than 30 is required for gram-positive organisms. Pharmacodynamic principles can also be used to devise the optimal administration regimen for specific antimicrobial agents. Pharmacodynamic analysis of the activity of levofloxacin against Streptococcus pneumoniae revealed that, 99% of the time, actual hospitalized patients achieve an AUC/MIC of greater than 30. This indicates that levofloxacin will be very effective in treating S. pneumoniae infections in the majority of patients. Cost of treatment is an increasing concern voiced by healthcare providers and users alike. This has led to a much greater emphasis placed upon the cost of individual drugs used in the management of infections. However, when evaluating the cost of an antibiotic, it is extremely important that not only are the direct acquisition costs assessed, but consideration also given to other aspects incurring a financial burden, such as drug preparation cost, supplies, costs of treating adverse events or any treatment failures. It is only by having such a full assessment of costs that realistic financial comparisons can be made between different antibiotics. PMID- 10810209 TI - Antimicrobial resistance in respiratory tract pathogens: results of an international surveillance study. AB - An international surveillance study was performed to assess the resistance patterns among respiratory tract pathogens during the winter of 1997-1998. The pathogens studied included Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. The antibiotics tested included five beta-lactams (penicillin, ampicillin, amoxicillin, amoxicillin-clavulanic acid, cefuroxime axetil and ceftriaxone), two macrolides (azithromycin and clarithromycin), one sulfonamide (trimethoprim-sulfamethoxazole), one glycopeptide (vancomycin) and one fluoroquinolone (levofloxacin). A total of 11,502 isolates were tested from nine countries, using microdilution susceptibility tests as recommended by National Committee for Clinical Laboratory Standards (NCCLS) guidelines. The susceptibility rates varied greatly around the world. Ninety percent of M. catarrhalis isolates produced beta-lactamase, making them resistant to ampicillin. beta-Lactamase production by H. influenzae ranged from 5% in Germany to 34% in the USA (mean 17.5%). Of the S. pneumoniae isolates, 32.8% had some resistance to penicillin, but this ranged greatly from 7.8% in Germany to 66.5% in France. Penicillin resistance in S. pneumoniae was associated with resistance to other beta-lactams, macrolides and sulfonamides, but not to levofloxacin or vancomycin. All isolates of H. influenzae and M. catarrhalis were susceptible to levofloxacin. Results of this study support the conclusion that these three respiratory tract pathogens are becoming more resistant to selected antimicrobials, and that the level of resistance in these isolates to the antimicrobials varies greatly from one country to another. PMID- 10810210 TI - Treatment of community-acquired pneumonia. AB - Lower respiratory tract infections are the major cause of death due to infectious disease in the developed and developing world. Despite substantial progress in defining pathogens and in therapeutic options, there continues to be major controversies in the clinical management of these infections. This report reviews the guidelines for community-acquired pneumonia from the Infectious Diseases Society of America (IDSA), updated from the initial publication. Diagnosis should include a chest X-ray to differentiate acute bronchitis from pneumonia. The decision for hospitalization should be based on social factors and evaluation of severity of illness. Identification of an etiological agent for inpatients should include two pretreatment cultures, one pretreatment sputum specimen, with seriously ill patients requiring studies for Legionella spp. Recommendations for empiric treatment of outpatients are doxycycline, a macrolide or a fluoroquinolone. Recommendations for empiric treatment of hospitalized patients are a cephalosporin plus a macrolide, or a fluoroquinolone alone. Recommendations for ICU patients are a beta-lactam combined with either a macrolide or a fluoroquinolone. While concern has arisen about increasing resistance to fluoroquinolones, arguments in favor of these agents include the fact that they have good in vitro activity against nearly all treatable pathogens except some anaerobes. Clinical trials have shown equivalence or superiority compared to other standard agents. They are well tolerated, and can be administered intravenously or orally, once daily. A recent retrospective review has shown superior outcome with fluoroquinolone treatment compared to cephalosporins, including a 36% reduction in mortality. PMID- 10810211 TI - Cytokine modulation in experimental endotoxemia: characterization of an ex vivo whole blood model. AB - A new model was developed to study cytokine regulation and modulation in whole blood ex vivo. The model is characterized by stable leukocyte counts and high leukocyte viability throughout the experimental period. Oxygen consumption per time decreased slowly, whereas carbon dioxide partial pressure increased accordingly throughout the experiment. In this model, the anti-inflammatory effects of recombinant human (rh) interleukin (IL)-4, rhIL-10 and rhIL-13 on lipopolysaccharide (LPS) stimulated (10 ng/ml) leukocytes were examined and compared by measuring their ability to inhibit the release and mRNA levels of tumor necrosis factor (TNF)alpha, IL-6 and IL-1beta. rhIL-10 potently inhibited the release of TNF-alpha, IL-6 and IL-1beta in a potent and dose-dependent manner, but did not influence the mRNA levels of these cytokines in CD14-positive cells. Also, rhIL-4 and rhIL-13 inhibited the release of IL-6 and IL-1beta in a potent and dose-dependent manner, however, stronger maximal inhibition of IL 1beta (85%) than of IL-6 (60%) was obtained. In contrast, rhIL-4 and rhIL-13 seemed to have both stimulatory and inhibitory effects on plasma values of TNF alpha. The effects of 10 ng/ml LPS showed to be signalling through the CD14 receptor, since blood treated with a monoclonal anti-CD14 antibody did not produce any TNF-alpha. The whole blood model described in this study is in our opinion a useful tool for investigating immunomodulating effects on a mixed white blood cell population. PMID- 10810212 TI - Effect of ibuprofen and diethylcarbamazine on the hemodynamic and inflammatory response to endotoxin in the dog. AB - We studied the effects of pretreatment with either ibuprofen (15 mg/kg), diethylcarbamazine (DEC, 15 mg/kg), or vehicle, on the hemodynamic, hematologic, and serum tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6 responses to a bolus Escherichia coli endotoxin infusion (2 mg/kg) in 21 pentobarbital-anesthetized dogs (n = 7 each group). Hematologic and cytokine data were collected before and 0.5, 1, and 3 h after endotoxin infusion. Endotoxin decreased arterial pressure (P(a)), cardiac output (CO), total leukocyte (WBC) and platelet counts, and increased lactate, TNF-alpha, and IL-6. TNF-alpha levels peaked at 1 h and decreased by 3 h, whereas IL-6 remained elevated. Ibuprofen abolished the endotoxin-induced changes in P(a) and lactate, but did not alter the initial decrease in CO, WBC, and platelets or the increase in TNF-alpha and IL-6. DEC did not alter the response to endotoxin, although the DEC group had higher TNF-alpha and IL-6 levels before endotoxin infusion compared to controls. We conclude that the cardiovascular effects of ibuprofen in the canine model of acute endotoxemia are not due to suppression of the systemic inflammatory response. PMID- 10810213 TI - Protective effect of human urinary thrombomodulin on ischemia- reperfusion injury in the canine liver. AB - This study was performed to determine whether human urinary soluble thrombomodulin plays a role in liver ischemia-reperfusion injury. Liver ischemia was induced in two groups of dogs. Group 1 was exposed to 60 min ischemia, and group 2 was exposed to 60 min ischemia after preischemic administration of human urinary soluble thrombomodulin. In group 1, the thrombin-antithrombin complex and hyaluronic acid were significantly elevated after ischemia, compared with the preischemic values. While liver issue blood flow and the plasmin-alpha(2)-plasmin inhibitor complex significantly decreased, AST, ALT and m-AST dramatically increased after reperfusion. In group 2, the increase in the thrombin antithrombin complex and hyaluronic acid was significantly suppressed, and AST, ALT and liver tissue blood flow significantly improved, compared with group 1. Histologically, in group 2, the hepatic tissue structure, including endothelial cells, was relatively intact. These findings suggest that administration of thrombomodulin inhibits endothelial cell injury and coagulopathy and offers protection from liver ischemia-reperfusion injury. PMID- 10810214 TI - Effect of preoperative portal vein embolization on liver volume and hepatic energy status of the nonembolized liver lobe in humans. AB - Clinically portal vein embolization (PVE) is presently preferred to extended hepatectomy. Nevertheless, its effect on hepatic adenosine triphosphate (ATP) and energy charge levels, which are essential for organ viability, has been little studied in humans. Fourteen patients with (n = 7) and without (n = 7) preoperative right PVE participated in this study. Changes in hepatic lobar volume and serum liver function tests were examined before and after percutaneous transhepatic right PVE. Liver volume (cm(3)) was calculated on computed tomograms before and 20+/-3 days after PVE. At the time of surgery (mean of 25 days after PVE), small liver specimens were obtained from portal vein (PV) nonembolized left lobes immediately after laparotomy without any ischemic procedures. Concentrations of adenine nucleotides were measured by high performance liquid chromatography, and hepatic energy charge levels were calculated. These values were compared with those in control patients who had not undergone preoperative PVE. Serum liver function tests including the indocyanine green retention rate did not differ significantly before and after PVE. The volume of the PV nonembolized left lobe significantly increased after right PVE (from 473+/-32 to 624+/-66 cm(3)), with a significant increase in the percentage of the left lobe to total liver volume. The concentrations of AMP, ADP, and ATP, and hepatic energy charge levels in the PV-nonembolized left lobe were similar to those of the control liver. These results suggest that preoperative right PVE increases the volume of the nonembolized left lobe, keeping the hepatic engery charge and ATP levels similar to the control liver, thereby increasing the total amount of ATP and hepatic energy reserve of the PV-nonembolized lobe in proportion to its volume increase at the time of surgery. PMID- 10810215 TI - Computer-assisted ex vivo, normothermic small bowel perfusion. AB - BACKGROUND: In the present study, a technique for computer-assisted, normothermic, oxygenated, ex vivo, recirculating small bowel perfusion was established as a tool to investigate organ pretreatment protocols and ischemia/reperfusion phenomena. A prerequisite for the desired setup was an organ chamber for ex vivo perfusion and the use of syngeneic whole blood as perfusate. METHODS: The entire small bowel was harvested from Lewis rats and perfused in an organ chamber ex vivo for at least 2 h. The temperature was kept at 37 degrees C in a water bath. Three experimental groups were explored, characterized by different perfusion solutions. The basic perfusate consisted of syngeneic whole blood diluted with either NaCl, Krebs' solution or Krebs' solution and norepinephrine to a hematocrit of 30%. In addition, in each group l-glutamine was administered intraluminally. The desired perfusion pressure was 100 mm Hg which was kept constant with a computer-assisted data acquisition software, which measured on-line pressure, oxygenation, flow, temperature and pH and adjusted the pressure by changing the flow via a peristaltic pump. The viability of the preparation was tested by measuring oxygen consumption and maltose absorption, which requires intact enzymes of the mucosal brush border to break down maltose into glucose. RESULTS: Organ perfusion in group 1 (dilution with NaCl) revealed problems such as hypersecretion into the bowel lumen, low vascular resistance and no maltose uptake. In contrast a viable organ could be demonstrated using Krebs' solution as dilution solution. The addition of norepinephrine led to an improved perfusion over the entire perfusion period. Maltose absorption was comparable to tests conducted with native small bowel. Oxygen consumption was stable during the 2-hour perfusion period. CONCLUSIONS: The ex vivo perfusion system established enables small bowel perfusion for at least 2 h. The viability of the graft could be demonstrated. The perfusion time achieved is sufficient to study leukocyte/lymphocyte interaction with the endothelium of the graft vessels. In addition, a viable small bowel, after 2 h of ex vivo perfusion, facilitates testing of pretreatment protocols for the reduction of the immunogenicity of small bowel allografts. PMID- 10810216 TI - Xenogeneic patch closure of the small intestine: a novel approach to fistula management. AB - Previous work from our laboratory demonstrated the feasibility of utilizing placental-derived collagen tissue matrix (CTM) as a bowel wall substitute. We reasoned that this technique would also be suitable in managing intestinal fistulae. To test this hypothesis, we created a chronic cecal fistula in rats and randomly managed some with primary repair and others with CTM replacement. Leak rates, mortality, bursting pressures and histologic scores were similar, suggesting that a chronic fistula can be successfully managed with either a CTM or primary repair. PMID- 10810217 TI - Effect of artificial valves on intestinal adaptation in the short-bowel syndrome: an integrated study of morphological and functional changes in rats. AB - Two-third-resections of the proximal or distal small bowel with and without artificial valves were performed in rats. Intestinal adaptation led to a significant increase in bowel diameter, villus height and villus diameter and consequently in absorptive mucosal surface area per unit of serosal area. Additional artificial valve construction did not affect the calculated mucosal surface area after proximal resection, while it significantly decreased the absorptive area by the occurrence of large, plump villi after distal resection. There was no change in small-intestinal absorption of water, glucose and electrolytes per unit mucosa with valve construction. DNA cytometry showed that artificial valves led to mucosal hyperplasia without hypertrophy. These morphological changes coincided with a significant increase in basal and stimulated gastrin release. The body weight was unchanged or even worse in the valve groups than after resection alone, despite a significantly prolonged transit time. Therefore, in our study, artificial valves did not result in functional improvements after small intestinal resections. PMID- 10810218 TI - Total pancreatico-duodenal transplantation with portal venous drainage: metabolic assessments in diabetic rats. AB - A perfect metabolic correction of diabetes is essential to completely eradicate long-term chronic complications. Only a total pancreatic graft with portal venous drainage enables such an achievement. Isogenic Lewis rats were used for donors, recipients and controls. Pancreatico-duodenal transplantation was either heterotopic with systemic venous drainage (n = 12) or paratopic with portal drainage (n = 11). All animals were regularly monitored for non-fasting plasma glucose and insulin. Both techniques promptly restored the non-fasting plasma glucose to normal values (p<0.003). Normo-insulinemia (47.4+/-6.4 microU/ml) was obtained in the paratopic group, while the heterotopic group showed hyperinsulinism (132.0+/-15.2 microU/ml). Perfect metabolic control justifies the additional technical difficulties of total paratopic pancreatic transplantation with portal venous drainage. PMID- 10810219 TI - Evaluation of seprafilm and amniotic membrane as adhesion prophylaxis in mesh repair of abdominal wall hernia in rats. AB - INTRODUCTION: Adhesion formation following abdominal wall hernia repair with prosthetic mesh may lead to intestinal obstruction and enterocutaneous fistula. Physical barriers, namely, human amniotic membrane (HAM) or Seprafilm (Genzyme, Cambridge, Mass., USA), a bio-absorbable, translucent membrane composed of carboxymethylcellulose and hyaluronic acid, have been reported to prevent postsurgical intra-abdominal adhesions. OBJECTIVE: Evaluating the effect of HAM and Seprafilm in preventing adhesion formation in the rat model of ventral hernia repair with polypropylene mesh (PPM). MATERIAL AND METHODS: Sixty female Sprague Dawley rats were divided into three groups. A full-thickness abdominal wall defect was created in each animal. Control animals had the PPM sutured into the defect, whereas in the other two groups, either HAM or Seprafilm were laid over the abdominal viscera before the repair with PPM. Half of the animals in each group were sacrificed on the 21st postoperative day. The remaining rats of the same group were re-operated on the 42nd day for investigation and measurement of the adhesion area in relation to the graft area. RESULTS: Direct mesh repair showed 52.8 and 56% area adhesion formation 3 and 6 weeks postoperatively, respectively. The HAM barrier covered with mesh repair demonstrated 0 and 0.96% area adhesion formation, and the Seprafilm-covered mesh repair showed 0 and 0% area adhesion formation 3 and 6 weeks postoperatively, respectively. Uncovered mesh showed a significantly larger adhesion area than both covered mesh (p = 0.001 and 0.001). Both HAM and Seprafilm were equally effective in preventing postoperative adhesions. CONCLUSIONS: HAM and Seprafilm proved to be an effective antiadhesive barrier in PPM repair of abdominal wall hernia. PMID- 10810220 TI - Preparation of a skeletal muscle ventricle in sheep: severe damage to the Latissimus dorsi muscle due to mobilization before preconditioning. AB - As part of a study examining the use of a skeletal muscle ventricle for cardiac assistance in sheep, a new concept of muscle preconditioning was put into practice. We aimed to produce a latissimus dorsi muscle (LDM) capable of performing chronic work immediately after the construction of a skeletal muscle ventricle. The left LDM was detached from the thoracic wall, divided longitudinally and reattached in situ to achieve vascular delay. The right LDM was left unaffected. Thereafter, preconditioning of both LDM was started according to the clinically approved stimulation protocol for cardiomyoplasty. Preconditioning of the unaffected right LDM in situ resulted in a complete muscle fiber transformation with no signs of degeneration or necrosis. Mobilization of the left LDM before preconditioning led to a distinct damage of the muscle. During conditioning, the increase in burst duration from 2 to 3 impulses in sheep A and from 3 to 5 impulses in sheep B resulted in a homogenous degeneration of the muscle fibers of the left LDM. Histomorphological analysis showed a dramatic increase in the percent perimysial and endomysial connective tissue. The applied concept of muscle prefabrication proved to be a failure. Muscle splitting and mobilization followed by vascular delay and in situ conditioning as a concept of muscle prefabrication should be strictly avoided. PMID- 10810221 TI - Conservation of plasma regulatory proteins of the complement system in evolution: humans and fish. AB - The complement system is an important defense system of innate immunity. The recent identification of structurally and functionally related complement regulatory proteins in the teleost, barred sand bass (Paralabrax nebulifer), and humans, two species which are separated in evolution by 100 million years, indicates a high level of conservation and the early presence of this defense system in evolution. The complement regulatory protein of barred sand bass, SBP1, is related to both the human alternative pathway regulator factor H, and to the classical pathway regulator C4bp, and displays regulatory activities in both human pathways. In addition, molecules with homology to the recently identified human factor-H-related proteins are also present in the sand bass genome. Here, we summarize the structural and functional aspects of these homologies and discuss the consequences for the evolution of the complement system. PMID- 10810222 TI - Childhood atopic asthma: positive association with a polymorphism of IL-4 receptor alpha gene but not with that of IL-4 promoter or Fc epsilon receptor I beta gene. AB - We examined the relative contributions of three representative candidate genes for atopy (Fc epsilon receptor I beta, IL-4, and IL-4 receptor alpha) to the development of atopic asthma. Four polymorphisms of the three candidate genes including Ile50Val and Gln551Arg of IL-4 receptor alpha, -590C/T of IL-4 promoter and Glu237Gly of Fc epsilon receptor I beta were studied in 100 patients with atopic asthma and 100 nonatopic controls in the northern Kyushu area in Japan. Among the four polymorphisms of the three candidate genes, the Ile50 allele of the IL-4 receptor alpha chain gene demonstrated an association with atopic asthma subjects (p = 0.044), especially in patients with onset at 2 years of age or earlier (p = 0.034) and in patients with moderate to severe atopic asthma (p = 0. 031). Gln551Arg of IL-4 receptor alpha, -590C/T of IL-4 promoter and Glu237Gly of Fc epsilon receptor I beta showed no association with atopic asthma. A slight linkage disequilibrium between Ile50Val and Gln551Arg polymorphisms of the IL-4 receptor alpha chain gene was observed in both patients and nonatopic controls. The identification of additional atopy genes in areas with a certain genetic background is essential for genetic diagnosis and to establish new therapeutic modalities for atopic asthma. PMID- 10810223 TI - Rapid purification of human DNase I using mouse monoclonal anti-DNase I antibodies and characterization of the antibodies. AB - Five anti-human deoxyribonuclease I (DNase I) monoclonal antibodies were obtained from BALB/c mice immunized with DNase I purified from human urine. Four of them inhibited DNase I enzyme activity, as did a rabbit polyclonal antibody; these 4 did not have immunostaining ability. The remaining one had immunostaining ability but no inhibitory activity. A Sepharose 4B column conjugated with 1 of the 4 antibodies that had inhibitory activity effectively adsorbed and eluted the DNase I enzyme; this did not occur with the rabbit polyclonal antibody. We showed that adding an immunoaffinity chromatography step made the purification of human DNase I easier and faster than the conventional procedure. PMID- 10810224 TI - Ontogeny recapitulates phylogeny: comparative immunology in Germany. AB - The innate immune system is in the spot light of modern immunology. Whenever protists, invertebrates and vertebrates are threatened by pathogens, they rapidly activate highly effective antimicrobial defense reactions. Because this young field develops very dynamically, it is important to ask what we really know about the mechanisms governing the innate immune defense system. This was the topic of a recent meeting entitled 'The Evolution of the Immune System', held at the Friedrich Schiller University in Jena, Germany. Leading scientists in the field of innate immunity presented their latest data in a historical and friendly setting. PMID- 10810225 TI - The human T cell receptor alpha variable (TRAV) genes. AB - 'Human T Cell Receptor Alpha Variable (TRAV) Genes', the eighth report of the 'IMGT Locus in Focus' section, comprises four tables: (1) 'Number of human germline TRAV genes at 14q11 and potential repertoire'; (2) 'Human germline TRAV genes at 14q11'; (3) 'Human TRAV allele table', and (4) 'Correspondence between the different human TRAV gene nomenclatures'. These tables are available at the IMGT Marie-Paule page of IMGT, the international ImMunoGeneTics database (http://imgt.cines.fr:8104) created by Marie-Paule Lefranc, Universite Montpellier II, CNRS, France. PMID- 10810226 TI - The human T cell receptor alpha joining (TRAJ) genes. AB - 'Human T cell Receptor Alpha Joining Genes', the 9th report of the 'IMGT Locus in Focus' section, comprises 3 tables: (1) 'Human germline TRAJ genes'; (2) 'Human TRAJ allele table'; and (3) 'Nucleotide and protein displays of the human TRAJ alleles (overview)'. These tables are available on the IMGT Marie-Paule page from IMGT, the international ImMunoGeneTics database (http://imgt. cines.fr:8104) created in 1989 by Marie-Paule Lefranc, Universite Montpellier II, CNRS, France. PMID- 10810227 TI - The human T cell receptor beta diversity (TRBD) and beta joining (TRBJ) genes. AB - 'Human T cell Receptor Beta Diversity (TRBD) and Beta Joining (TRBJ) Genes', the 10th report of the 'IMGT Locus in Focus' section, comprises 6 tables: (1) 'Human germline TRBD genes'; (2) 'Human TRBD allele table'; (3) 'Nucleotide and protein displays of the human TRBD alleles (overview)'; (4) 'Human germline TRBJ genes'; (5) 'Human TRBJ allele table', and (6) 'Nucleotide and protein displays of the human TRBJ alleles (overview)'. These tables are available on the IMGT Marie Paule page from IMGT, the international ImMunoGeneTics database (http://imgt.cines.fr:8104) created in 1989 by Marie-Paule Lefranc, Universite Montpellier II, CNRS, France. PMID- 10810228 TI - The molecular basis of cystinuria: an update. AB - Cystinuria is a hereditary disorder of cystine and dibasic amino acid transport across the luminal membrane of renal proximal tubule and small intestine. In 1992, a cDNA (rBAT) was isolated from kidney which induced high-affinity, sodium independent uptake of cystine and dibasic amino acids when expressed in Xenopus oocytes. The rBAT gene was mapped to a region of chromosome 2p known to contain a cystinuria locus, and rBAT expression was demonstrated in the straight (S3) portion of renal proximal tubule and small intestine. Over 30 distinct rBAT mutations have been described in patients who inherit two fully recessive (type I) cystinuria genes. Recently, the second cystinuria gene (SLC7A9) on chromosome 19q was identified; SLC7A9 mutations were shown to cause the incompletely recessive form of cystinuria (types II and III). Patients who inherit two mutant SLC7A9 genes have recurrent nephrolithiasis comparable to those with two rBAT mutations. In some cystinuria families, patients inherit a fully recessive allele from one parent and an incompletely recessive allele from the other parent; patients with this 'mixed type' of cystinuria have somewhat milder disease. It is not yet clear whether this form of cystinuria involves rBAT as well as SLC7A9 mutations. Current evidence suggests that the transmembrane channel mediating uptake of cystine and dibasic amino acids at the luminal surface is encoded by SLC7A9; the smaller rBAT protein forms a heterodimeric complex with this channel and is critical for its targetting to the luminal membrane. PMID- 10810229 TI - Arginase in glomerulonephritis. AB - Arginase metabolizes L-arginine to L-ornithine and urea. Two arginase isoforms, AI (liver arginase) and AII (ubiquitously expressed, functions unknown), have been identified. It is clear that arginases potentially have important roles in addition to urea generation for high concentrations are present at inflammatory sites. Regulation occurs through cytokines, substrate competition and products of nitric oxide (NO) metabolism. The functions of arginases at inflammatory sites are unknown, but may include regulation of apoptosis and NO production and generation of structural and cellular protein precursors. In glomerulonephritis there is increased arginase activity in nephritic glomeruli following a pattern similar to that in wound healing. The level can be further increased by NO inhibition suggesting substrate competition. The potential sources in the inflamed glomerulus include infiltrating leucocytes and mesangial cells, and the predominant isoform expressed is AI (AII predominates under physiological conditions). The recent identification of different isoforms of arginase has been an important step towards understanding the significance of arginase activity in glomerulonephritis. PMID- 10810231 TI - Inhibition with antisense oligonucleotide suggests that IkappaB-alpha does not form a negative autoregulatory loop for NF-kappaB in mesangial cells. AB - The IkappaB proteins are important in the regulation of the NF-kappaB/Rel group of transcription factors which are pivotal in the inflammatory response. IkappaB alpha is itself upregulated by activation of NF-kappaB and is postulated to be part of a negative feedback loop. This role of IkappaB-alpha has been challenged, however, by recent evidence that demonstrates (1) continued activation of NF kappaB in mesangial and endothelial cells despite the resynthesis of IkappaB alpha protein and (2) that inhibition of the transactivating activity of NF kappaB by corticosteroids can be dissociated from a rise in IkappaB-alpha protein. We investigated the role of IkappaB-alpha in mesangial cells using a phosphorothioate antisense oligonucleotide directed against the translational start point of IkappaB-alpha. If IkappaB-alpha does function as a negative feedback inhibitor in these cells, then reducing IkappaB-alpha levels should lead to an increase in NF-kappaB activity. We first demonstrated that IkappaB-alpha protein resynthesis following stimulation could be specifically reduced. We then showed that NF-kappaB DNA binding was not increased with antisense treatment following stimulation. Finally, NF- kappaB-dependent gene signalling after stimulation (determined through an NF-kappaB luciferase reporter and upregulation of the mRNA of known NF-kappaB-responsive genes MCP-1 and IkappaB-alpha) was reduced rather than increased. These data suggest that IkappaB-alpha does not form a negative autoregulatory loop for NF-kappaB in mesangial cells and may actually reduce NF-kappaB activity. This may have relevance to therapies directed at inhibition of NF-kappaB activity in mesangial cell diseases. PMID- 10810230 TI - Inhibiting albumin glycation ameliorates diabetic nephropathy in the db/db mouse. AB - Albumin modified by Amadori glucose adducts stimulates the expression of extracellular matrix proteins by glomerular mesangial and endothelial cells, and has been mechanistically linked to the pathogenesis of diabetic nephropathy. To test the hypothesis that inhibiting the formation of glycated albumin might beneficially influence the development of kidney disease in diabetes, we treated diabetic db/db mice for 12 weeks with a low-molecular-weight compound (EXO-226) that impedes the condensation of free glucose with lysine epsilon-amino groups in albumin. Administration of EXO-226 (3 mg/kg) twice daily by gavage normalized the plasma concentration of glycated albumin within days after initiation of treatment and maintained glycated albumin within the normal range throughout the study, despite persistent and severe hyperglycemia. Urine albumin excretion, which was markedly increased at the start of the study (age 12 weeks), was significantly reduced in treated diabetic animals compared with their untreated diabetic littermates. The fall in creatinine clearance that was observed in untreated diabetic animals was prevented in diabetic littermates that received treatment. Compared with the nondiabetic controls, the amount of glomerular mesangial matrix was threefold greater in untreated diabetic mice; in contrast, the mesangial matrix fraction was only 1. 5 times that of nondiabetic controls in the treated diabetic animals, representing a reduction in mesangial matrix accumulation of more than 50%. EXO-226 also reduced the overexpression of mRNA encoding for alpha1 (IV) collagen in renal cortex of db/db mice. We conclude that normalization of plasma glycated albumin concentrations with the glycation inhibitor EXO-226 ameliorates the glomerular structural and functional abnormalities associated with diabetic nephropathy in the db/db mouse. PMID- 10810232 TI - Cloning of rabbit Cct6 and the distribution of the Cct complex in mammalian tissues. AB - Cct6 protein is one of the subunits of the Cct complex involved in ATP-dependent folding of cellular proteins. We used the cDNA of the human CCT6 subunit to obtain a full-length cDNA from a rabbit kidney cortex library. Two transcripts of 2 and 2.5 kb were detected in rabbit kidney and liver by Northern analysis. The rabbit CCT6 was 93% identical to the human gene; the deduced amino acid sequence was 97% identical. A phylogenetic analysis of Cct6 proteins from mouse, rabbit, human, and yeast showed greater similarities of Cct6 protein among the species than among other Cct subunits. The ATP-binding sites were perfectly conserved among mammals and yeast, supporting the role of Cct complex in ATP-dependent protein folding. Using a polyclonal antibody to human Cct6 protein and Western analysis, we found expression of this subunit in a variety of rabbit organs and tissues, as well as in bovine testes, human lymphocytes, human and rabbit reticulocytes, and in two cultured kidney cell lines. We also found Cct1 protein by Western analysis in several rabbit organs as well as in bovine testes. These data characterize the rabbit Cct6 subunit and compare it to its homologues. The Western analyses support the concept that Cct complex is widely distributed among tissues and highly conserved among eukaryotes. PMID- 10810233 TI - Role of intrarenal endothelin 1, endothelin 3, and angiotensin II expression in chronic cyclosporin A nephrotoxicity in rats. AB - Endothelin 1 (Et1) is widely expressed in the kidney and is related to several functions and to pathological conditions with progression towards sclerosis. The function of endothelin 3 (Et3) at the renal level is debatable, but it could have an important regulatory function in the reabsorption of water through its action on tubular type B receptors. Angiotensin II has recently been implicated as the principal factor responsible for the progression of interstitial fibrosis induced by cyclosporin A (CsA). We investigated this relationship in vivo and analyzed the modifications induced by CsA toxicity in Sprague-Dawley rats treated with 25 mg/kg/day of CsA for 28 and 56 days. Immunohistochemical methods and molecular analysis were used to study the expression of Et1 and Et3 and immunohistochemistry alone to determine the intrarenal expression of angiotensin II. Rats treated with CsA developed chronic nephrotoxicity lesions; semiquantitative analyses of hyaline arteriolopathy revealed that the passage of time affected the extent of this lesion and led to the diminution of the total glomerular area. Immunohistochemical results showed that chronic CsA treatment induced moderate secretion of Et1 and Et3 at tubular and glomerular levels and that the local expression of angiotensin II in the treatment groups was more evident than in control animals. Besides, the mRNA levels of preproEt3 showed a dramatic increase from 28 days after CsA treatment (control group 0.07+/-0.11 vs. CsA group 0.48+/-0.11, p<0.01), while the mRNA levels of preproEt1 increased from 56 days (control group 0.15+/-0.05 vs. CsA group 0.34+/-0.09, p< 0.05). At 28 days, renal lesions correlated strongly with the mRNA levels of Et3 (r>0.50, p<0.01). However, at 56 days, the key finding was the strong correlation of the most important analytical, histological, and immunohistochemical parameters of CsA nephrotoxicity with Et1 mRNA levels (r>0.50, p<0.01). These results support the hypothesis that the clinical and morphological phenomena linked with CsA nephrotoxicity are related to hypersecretion of endothelins and local expression of angiotensin II in the outer medulla and medullary rays; Et3 and angiotensin II are the first to act, followed subsequently by Et1. PMID- 10810235 TI - Overview. PMID- 10810234 TI - Preferential COX-2 inhibitor, meloxicam, compromises renal perfusion in euvolemic and hypovolemic rats. AB - Nonsteroidal anti-inflammatory drugs can impair renal perfusion through inhibition of cyclooxygenase (COX)-mediated prostaglandin synthesis. We investigated the influence of the preferential COX-2 inhibitor, meloxicam (MELO), on renal hemodynamics in eu- and hypovolemic rats compared to the nonselective COX inhibitor indomethacin (INDO). The hypovolemic state was obtained in rats by three daily injections of furosemide (2 mg/kg i.p.) followed by a sodium deficient diet for 7 days. In euvolemic rats (n = 6) neither INDO (5 mg/kg i.v.) nor MELO (1 or 2 mg/kg i.v.) influenced mean arterial blood pressure (MAP) or impaired renal (RBF) and cortical blood flow (CBF). Medullary blood flow (MBF) decreased after INDO (18%; p<0.05), and dose-dependently after MELO (1 mg, 10%; 2 mg, 18%; p<0.05). In hypovolemic rats (n = 6) INDO and MELO had no effect on MAP. RBF and CBF were reduced after INDO (11 or 20%; p<0. 05), but showed no changes after MELO. INDO induced a decrease in MBF (22%; p<0.05) which was less pronounced after MELO (12%; p <0.05). In conclusion the preferential COX-2 inhibitor MELO compromized renal perfusion in the outer medulla both in eu- and hypovolemic animals. PMID- 10810236 TI - Supraspinal circuitry mediating opioid antinociception: antagonist and synergy studies in multiple sites. AB - Supraspinal opioid antinociception is mediated by sensitive brain sites capable of supporting this response following microinjection of opioid agonists. These sites include the ventrolateral periaqueductal gray (vIPAG), the rostral ventromedial medulla (RVM), the locus coeruleus and the amygdala. Each of these sites comprise an interconnected anatomical and physiologically relevant system mediating antinociceptive responses through regional interactions. Such interactions have been identified using two pharmacological approaches: (1) the ability of selective antagonists delivered to one site to block antinociception elicited by opioid agonists in a second site, and (2) the presence of synergistic antinociceptive interactions following simultaneous administration of subthreshold doses of opioid agonists into pairs of sites. Thus, the RVM has essential serotonergic, opioid, cholinergic and NMDA synapses that are necessary for the full expression of morphine antinociception elicited from the vIPAG, and the vIPAG has essential opioid synapses that are necessary for the full expression of opioid antinociception elicited from the amygdala. Further, the vIPAG, RVM, locus coeruleus and amygdala interact with each other in synergistically supporting opioid antinociception. PMID- 10810237 TI - Delta opioid peptide[D- Ala(2),D-Leu(5)]enkephalin promotes cell survival. AB - By studying the hibernation in ground squirrels, a protein factor termed hibernation induction trigger (HIT) was found to induce hibernation in summer active ground squirrels. Further purification of HIT yielded an 88-kD peptide that is enriched in winter hibernator. Partial sequence of the 88-kD protein indicates that it may be related to the inhibitor of metalloproteinase. Delta opioid [D-Ala(2),D-Leu(5)]enkephalin (DADLE) also induced hibernation. HIT and DADLE were found to prolong survival of peripheral organs preserved en bloc or as a single preparation. These organs include the lung, the heart, liver and kidney. DADLE also promotes survival of neurons in the central nervous system. Methamphetamine (METH) is known to cause destruction of dopaminergic (DA) terminals in the brain. DADLE blocked and reversed the DA terminal damage induced by METH. DADLE acted against this effect of METH at least in part by attenuating the mRNA expressions of a tumor necrosis factor p53 and an immediate early gene c fos. DADLE also blocked the neuronal damage induced by ischemia-reperfusion following a transient middle cerebral artery occlusion. In PC12 cells, DADLE blocked the cell death caused by serum deprivation in a naltrexone-sensitive manner. Thus, DADLE, and by extension the endogenous delta opioid peptides and delta opioid receptors, may play an important role in organ and neuronal survival. Here, critical developments concerning these fascinating cell protective properties of DADLE are reviewed. PMID- 10810238 TI - Spinal plasticity of acute opioid tolerance. AB - Spinal acute opioid tolerance remains mechanistically undercharacterized. Expanded clinical use of direct spinal administration of opioids and other analgesics indicates that studies to further understand spinal mechanisms of analgesic tolerance are warranted. Rodent models of spinal administration facilitate this objective. Specifically, acute spinal opioid tolerance in mice presents a plasticity-dependent, rapid, and efficient opportunity for evaluation of novel clinical agents. Similarities between the pharmacology of acute and chronic spinal opioid tolerance, neuropathic pain, and learning and memory suggest that this model may serve as a high through-put predictor of bioactivity of novel plasticity-modifying compounds. PMID- 10810239 TI - Endomorphins: localization, release and action on rat dorsal horn neurons. AB - Endomorphin (Endo) 1 and 2, two tetrapeptides isolated from the bovine and human brain, have been proposed to be the endogenous ligand for the mu-opiate receptor. A multi-disciplinary study was undertaken to address the issues of localization, release and biological action of Endo with respect to the rat dorsal horn. First, immunohistochemical studies showed that Endo-1- or Endo-2-like immunoreactivity (Endo-1- or Endo-2-LI) is selectively expressed in fiber-like elements occupying the superficial layers of the rat dorsal horn, which also exhibit a high level of mu-opiate receptor immunoreactivity. Second, release of immunoreactive Endo-2 like substances (irEndo) from the in vitro rat spinal cords upon electrical stimulation of dorsal root afferent fibers was detected by the immobilized antibody microprobe technique. The site of release corresponded to laminae I and II where the highest density of Endo-2-LI fibers was localized. Lastly, whole cell patch clamp recordings from substantia gelatinosa (SG) neurons of rat lumbar spinal cord slices revealed two distinct actions of exogenous Endo-1 and Endo-2: (1) depression of excitatory and/or inhibitory postsynaptic potentials evoked by stimulation of dorsal root entry zone, and (2) hyperpolarization of SG neurons. These two effects were prevented by the selective mu-opiate receptor antagonist beta-funaltrexamine. The localization of endomorphin-positive fibers in superficial layers of the dorsal horn and the release of irEndo upon stimulation of dorsal root afferents together with the observation that Endo inhibits the activity of SG neurons by interacting with mu-opiate receptors provide additional support of a role of Endo as the endogenous ligand for the mu-opiate receptor in the rat dorsal horn. PMID- 10810240 TI - G protein activation by endomorphins in the mouse periaqueductal gray matter. AB - The midbrain periaqueductal gray matter (PAG) is an important brain region for the coordination of mu-opioid-induced pharmacological actions. The present study was designed to determine whether newly isolated mu-opioid peptide endomorphins can activate G proteins through mu-opioid receptors in the PAG by monitoring the binding to membranes of the non-hydrolyzable analog of GTP, guanosine-5'-O-(3 [(35)S]thio)triphosphate ([(35)S]GTPgammaS). An autoradiographic [(35)S]GTPgammaS binding study showed that both endomorphin-1 and -2 produced similar anatomical distributions of activated G proteins in the mouse midbrain region. In the mouse PAG, endomorphin-1 and -2 at concentrations from 0.001 to 10 microM increased [(35)S]GTPgammaS binding in a concentration-dependent manner and reached a maximal stimulation of 74.6+/-3.8 and 72.3+/-4.0%, respectively, at 10 microM. In contrast, the synthetic selective mu-opioid receptor agonist [D-Ala(2),NHPhe(4), Gly-ol]enkephalin (DAMGO) had a much greater efficacy and produced a 112.6+/-5.1% increase of the maximal stimulation. The receptor specificity of endomorphin stimulated [(35)S]GTPgammaS binding was verified by coincubating membranes with endomorphins in the presence of specific mu-, delta- or kappa-opioid receptor antagonists. Coincubation with selective mu-opioid receptor antagonists beta funaltrexamine or D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Phe-Thr-NH(2) (CTOP) blocked both endomorphin-1 and-2-stimulated [(35)S]GTPgammaS binding. In contrast, neither delta- nor kappa-opioid receptor antagonist had any effect on the [(35)S]GTPgammaS binding stimulated by either endomorphin-1 or -2. These findings indicate that both endomorphin-1 and -2 increase [(35)S]GTPgammaS binding by selectively stimulating mu-opioid receptors with intrinsic activity less than that of DAMGO and suggest that these new endogenous ligands might be partial agonists for mu-opioid receptors in the mouse PAG. PMID- 10810241 TI - Actions of endomorphins on synaptic transmission of Adelta-fibers in spinal cord dorsal horn neurons. AB - The effects of endogenous mu-opioid ligands, endomorphins, on Adelta-afferent evoked excitatory postsynaptic currents (EPSCs) were studied in substantia gelatinosa neurons in spinal cord slices. Under voltage-clamp conditions, endomorphins blocked the evoked EPSCs in a dose-dependent manner. To determine if the block resulted from changes in transmitter release from glutamatergic synaptic terminals, the opioid actions on miniature excitatory postsynaptic currents (mEPSCs) were examined. Endomorphins (1 microM) reduced the frequency but not the amplitude of mEPSCs, suggesting that endomorphins directly act on presynaptic terminals. The effects of endomorphins on the unitary (quantal) properties of the evoked EPSCs were also studied. Endomorphins reduced unitary content without significantly changing unitary amplitude. These results suggest that in addition to presynaptic actions on interneurons, endomorphins also inhibit evoked EPSCs by reducing transmitter release from Adelta-afferent terminals. PMID- 10810242 TI - Pharmacological characterization of the nociceptin receptor, ORL1. Insight from the inward rectifier activation in the periaqueductal gray. AB - A novel opioid receptor-like orphan receptor (ORL1) was cloned and identified to be homologous to classical opioid receptors but insensitive to traditional opioids. A heptadecapeptide, termed orphanin FQ or nociceptin (OFQ/N), was identified as its endogenous ligand. OFQ/N shares overlapping distribution sites in pain-processing areas and common cellular mechanisms with opioids but exerts diverse effects on nociceptive responses. Of the two reported ORL1 antagonists, [Phe(1)psi(CH(2)-NH)- Gly(2)] nociceptin-(1-13)-NH(2) (Phepsi) and naloxone benzoylhydrazone (NBZ), antagonisms were validated in the activation of inward rectifying K channels induced by OFQ/N, using the patch clamp technique in ventrolateral periaqueductal gray slices. Results showed that Phepsi acted as a partial agonist and NBZ was a weak nonselective antagonist of ORL1. It is comparable with most but not all of the findings from other tissues. Comparing all the reports supports the above inference for these two antagonists. The possible causes for the discrepancy were discussed. A brief review on the putative ORL1 antagonists, acetyl-RYYRIK-NH2, some sigma-ligands and the functional antagonist, nocistatin, is also included. It indicates that a potent and selective ORL1 antagonist is expecting to elucidate the physiological role of OFQ/N. PMID- 10810243 TI - Reduction of lipopolysaccharide-induced neurotoxicity in mouse mixed cortical neuron/glia cultures by ultralow concentrations of dynorphins. AB - Previously we reported that ultralow concentrations of dynorphins (10(-16) to 10( 12) M) inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and proinflammatory cytokines in mouse glia without the participation of kappa opioid receptors. In the current study using mouse cortical neuron-glia cocultures, we examined the possibility that inhibition of glia inflammatory response by dynorphins might be neuroprotective for neurons. LPS, in a concentration-dependent manner, markedly increased the release of lactate dehydrogenase (LDH), an indicator of cellular injury. Ultralow concentrations (10(-14) to 10(-12) M) of dynorphin (dyn) A-(1-8) significantly prevented the LPS induced release of LDH, loss of neurons, and changes in cell morphology, in addition to inhibition of LPS-induced nitrite production. Meanwhile, ultralow concentrations (10(-15) to 10(-13) M) of des-[Tyr(1)]-dyn A-(2-17), a nonopioid peptide which does not bind to kappa-opioid receptors, exhibited the same inhibitory effect as dyn A-(1-17). These results suggest that dynorphins at ultralow concentrations are capable of reducing LPS-induced neuronal injury and these neuroprotective effects of dynorphins are not mediated by classical opioid receptors. PMID- 10810244 TI - Itch-scratch responses induced by opioids through central mu opioid receptors in mice. AB - We examined scratch-inducing effects of intracisternal, intrathecal and intradermal injections of morphine and some opioid agonists in mice. Intracisternal injection of morphine (3 nmol/animal) and the mu-receptor agonist [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]enkephalin (DAMGO; 0.2 nmol/animal) elicited scratching of the face, with little effect on scratching of the trunk. Intracisternal injection of the delta-receptor agonist [D-Pen(2,5)]enkephalin (DPDPE) and the kappa-receptor agonist U50488 were without effects. Intrathecal injection of morphine (0.1-3 nmol/animal) produced a dose-dependent increase in body scratching, with little effects on face scratching. Face scratching induced by intrathecal morphine (3 nmol/animal) was almost abolished by subcutaneous pretreatment with naloxone (1 mg/kg). Intradermal injections of morphine (3-100 nmol/site), DAMGO (1-100 nmol/site), DPDPE (10 and 100 nmol/site) and U50488 (10 100 nmol/site) did not elicit scratching of the site of injection. Intradermal injection of histamine (100 nmol/site) induced the scratching in ICR, but not ddY, mice and serotonin (30 and 50 nmol/site) elicited the scratching in either strain of mice. The results suggest that opioids induce scratching, and probably itching, through central mu-opioid receptors in the mouse. PMID- 10810245 TI - Effect of prenatal opioid exposure on cholinergic development. AB - Opioid drugs such as methadone or buprenorphine are often used in the management of pregnant addicts. These drugs are generally thought of as nonteratogenic and preferable to repeated cycles of withdrawal in utero. However, evidence exists that perinatal exposure to these opioids delays and disrupts cholinergic development, particularly in the striatum. Acetylcholine (ACh) content and the expression of choline acetyltransferase protein and mRNA are reduced in the early postnatal period by prenatal opioid exposure in the rat. Although these indicators of the cholinergic phenotype return to normal levels over time, the activity of the cholinergic neurons remains disrupted, with a large increase in ACh turnover rate. The mechanism of these effects is unknown, but may involve changes in the expression of nerve growth factor, which is reduced by opioid exposure. PMID- 10810246 TI - Acute cross-tolerance to opioids in heroin delta-opioid-responding Swiss Webster mice. AB - It is generally thought that the mu receptor actions of metabolites, 6 monoacetylmorphine (6MAM) and morphine, account for the pharmacological actions of heroin. However, upon intracerebroventricular (i.c.v.) administration in Swiss Webster mice, heroin and 6MAM act on delta receptors while morphine acts on mu receptors. Swiss Webster mice made tolerant to subcutaneous (s.c.) morphine by morphine pellet were not cross-tolerant to s.c. heroin (at 20 min in the tail flick test). Now, opioids were given in combination, s.c. (6.5 h) and i.c.v. (3 h) preceding testing the challenging agonist i.c.v. (at 10 min in the tail flick test). The combination (s.c. + i.c.v.) morphine pretreatment induced tolerance to the mu action of morphine but no cross-tolerance to the delta action of heroin, 6MAM and DPDPE and explained why morphine pelleting did not produce cross tolerance to s.c. heroin above. Heroin plus heroin produced tolerance to delta agonists but not to mu agonists. Surprisingly, all combinations of morphine with the delta agonists produced tolerance to morphine which now acted through delta receptors (inhibited by i.c.v. naltrindole), an unusual change in receptor selectivity for morphine. PMID- 10810248 TI - The role of local induction of tumor necrosis factor by LPS within a subcutaneous air pouch in the development of a febrile response in guinea pigs. AB - In rats, fever can be induced by injection of bacterial lipopolysaccharide (LPS) into a subcutaneous air pouch. This febrile response is in part dependent on the local action of interleukin-1beta within the pouch. In the present study, we tried to find out if this model of fever induction can be applied in guinea pigs and if the local LPS-induced formation of tumor necrosis factor-alpha (TNF-alpha) participates in the development of the febrile response. A dose of 100 microg/kg LPS was injected into a subcutaneous air pouch along with solvent (0.9% saline), or 100 mg/kg pentoxifylline, or 1 mg/animal soluble 55-kD TNF receptor (referred to as TNF-binding protein, TNF bp). The mean LPS-induced concentration of TNF in the lavage of the air pouch (measured 60 min after injection) was 17,765 pg/ml in animals injected with LPS and solvent. This value was reduced to 7,631 pg/ml if pentoxifylline was injected along with LPS into the pouch. If LPS was injected along with TNF bp, no bioactive TNF was detected in the lavage of the air pouch. Simultaneously, TNF was measured in blood plasma. Circulating concentrations of TNF were about 5-8% of the values detected in the lavage of the air pouch (mean values: 1,366 pg/ml in response to LPS plus solvent; 377 pg/ml in response to LPS plus pentoxifylline; no circulating TNF at all in response to LPS plus TNF bp). These data indicated a small spill-over of TNF from the air pouch into the circulation. Neither the reduction of local TNF formation in the air pouch by pentoxifylline nor the complete neutralization of TNF within the pouch by TNF bp resulted in a significant attenuation of the febrile response induced by injection of LPS into the subcutaneous air pouch. If there were an activation of cutaneous nerves to induce fever by a local formation of cytokines within the air pouch, TNF would not represent a likely candidate to be responsible for such a neural stimulation. PMID- 10810247 TI - The nucleus paragigantocellularis and opioid withdrawal-like behavior. AB - Participation of the nucleus paragigantocellularis (PGi) in mediation of opioid withdrawal was examined in conscious, unrestrained, non-opioid-dependent rats, using electrical stimulation of the PGi. A characteristic series of behaviors, which resembled those seen during naloxone-precipitated withdrawal from dependence on the opioid agonist, butorphanol, was elicited during 30 min of PGi stimulation. Thus, the behavioral syndrome has been termed opioid withdrawal like. Simultaneous microdialysis measurement of glutamate within the locus ceruleus indicated a positive correlation between extracellular glutamate concentrations and behavioral responses. Behavioral responses were inhibited by 50% during reverse dialysis perfusion of the locus ceruleus with the glutamate receptor antagonist, kynurenic acid, without any effect on glutamate concentrations. Thus, increases in locus ceruleus glutamate partially mediate opioid withdrawal-like behavior. Intracerebroventricular (i.c.v.) injections of the opioid antagonist, naloxone, or of the mu-selective (beta-funaltrexamine) or the delta-selective (naltrindole) opioid antagonists decreased, but did not abolish, stimulation-induced behavioral responses. Similar i.c.v. injections of the kappa-selective antagonist, nor-binaltorphimine, had no effect on behavioral responses to PGi stimulation. Activation of the PGi by electrical stimulation can elicit behaviors similar to those observed during opioid withdrawal. Moreover, additional levels of complexity are evident in the neuropharmacology of PGi stimulation-induced opioid withdrawal-like behavior. PMID- 10810249 TI - Circulating interleukin-1-beta levels after acute and prolonged exposure to low temperatures: human and rat studies. AB - In this study we have investigated whether IL-1 acts as a mediator of stress responses elicited by exposure to low temperatures. We also sought whether IL-1 is released from the adrenal gland under basal conditions or after exposure to low temperatures. Normal and adrenalectomized (ADX) rats were used for acute studies, whereas the effects of a prolonged exposure were investigated in a group of human subjects during a 45-day stay in Antarctica. Circulating levels of interleukin-1beta (IL-1beta) were taken as a marker of systemic IL-1 production both in humans and rats. In the latter, serum corticosterone (Cort) was also estimated. In intact rats, exposure to low temperatures (-25 or -35 degrees C) for 30 or 90 min did not modify circulating IL-1beta levels with respect to controls taken at +20 degrees C. Adrenalectomy was associated with an increase in cytokine levels only in the group exposed to -35 degrees C for 90 min; such increase is statistically significant compared to all groups of normal rats, whatever the experimental condition, as well as to ADX rats exposed to +20 degrees C and -25 degrees C for 30 and 90 min. In normal rats, the increase in circulating Cort levels was already maximal after exposure to -25 degrees C for 30 min. In humans, circulating IL-1beta levels after 45 days in Antarctica were significantly lower than those measured on arrival in the same subjects. Thus, no change in circulating IL-1beta was associated with acute low-temperature stress in rats, whereas a marked decrease in serum cytokine was observed in humans after prolonged exposure to a cold environment. Experiments with ADX rats indicated that the contribution of the adrenal glands to total-body IL-1beta production is negligible or absent. PMID- 10810250 TI - Activation of the adrenocortical axis by surgical stress: involvement of central norepinephrine and interleukin-1. AB - In the present study, we examined the mechanisms involved in the activation of the adrenocortical axis following surgical stress. Adult male rats underwent surgical laparotomy or craniotomy under ether anesthesia while control rats were only ether-anesthetized. Four hours following laparotomy or craniotomy, serum adrenocorticotropin (ACTH) and corticosterone (CS) were significantly increased and returned to almost basal levels after 24 h. Laparotomy also caused a significant depletion of corticotropin-releasing hormone (CRH-41) in the median eminence (ME). Pretreatment with dexamethasone 50 microg/kg completely abolished the pituitary-adrenal response while pretreatment with type II corticosteroid receptor antagonist caused a significant hypersecretion of both ACTH and CS and inhibited the effect of dexamethasone. The response to laparotomy was markedly attenuated in rats injected with 6-hydroxydopamine into the paraventricular nucleus (PVN) which significantly depletes norepinephrine (NE) PVN content. Intracerebroventricular injection of interleukin-1 receptor antagonist (IL-1ra) also inhibited the pituitary-adrenal response to laparotomy. The results suggest that (1) surgical stress activates the hypothalamo-pituitary-adrenal (HPA) axis via a mechanism which involves the release of CRH from the ME and NE input to the PVN; (2) the adrenocortical response is sensitive to the negative feedback of glucocorticoids via the mediation of type II glucocorticoid receptors, and (3) central IL-1 may be a mediator in the HPA axis response to surgical stress. PMID- 10810251 TI - Activation of the hypothalamic-pituitary-adrenal axis in IL-1 beta-converting enzyme-deficient mice. AB - Interleukin-1beta (IL-1beta) plays a key role in immune, behavioral and neuroendocrine responses to inflammation or infection. IL-1beta could also be involved in the response of the hypothalamic-pituitary-adrenal (HPA) axis during stress. Mature IL-1beta derives from a 31-kD precursor (pro-IL-1beta) that is processed by IL-1beta-converting enzyme (ICE). Mice in which the ICE gene has been nullated by homologous recombination were used to investigate the role of IL 1beta in the HPA axis response. Plasma levels of corticosterone and adrenocorticotropic hormone (ACTH) in response to an intraperitoneal injection of 5 microg lipopolysaccharide (LPS) were similar in ICE-deficient mice and wild type (WT) controls. In contrast, plasma ACTH response to restraint or to 200 ng of rat recombinant IL-1beta (rrIL-1beta) was higher in ICE-deficient mice as compared to WT animals. This hyperreactivity of the HPA axis in ICE knockout mice appears not to be related to the production of plasma IL-1beta or IL-6, which was similar to that of WT mice after rrIL-1beta injection. After lipopolysaccharide, ICE-deficient mice exhibited a smaller increase in plasma-immunoreactive IL-1beta and IL-6 as compared to WT controls. After restraint stress neither increase in plasma IL-1beta nor IL-6 was observed. The mechanisms responsible for the increased reactivity of the HPA axis in ICE-deficient mice may result from a higher sensitivity of the HPA axis to inflammatory cytokines or to cleavage products of pro-IL-1beta processed by non-ICE proteases. PMID- 10810253 TI - Fifth annual brain immune network group meeting. the jenner centre, bristol royal infirmary, bristol, uk, november 5, 1999 PMID- 10810252 TI - Effect of spleen-cell-conditioned medium on [3H]-choline uptake by retinal cells in vitro is mediated by IL-2. AB - Cytokines are essential molecules throughout the development of the nervous system and also play an important role during the adult life span. In the present work, we analyzed in vitro the effect of spleen-cell-conditioned medium (SCM) on the survival and [3H]-choline uptake of neonatal rat retinal cells. SCM induced an increase in neuronal survival, glial cell proliferation and neurite outgrowth, as evaluated by biochemical and morphological criteria. These effects were time dependent; after 120 h, SCM induced a 6-fold increase in the total protein level. The effect of SCM was blocked both by the inhibition of protein tyrosine kinase activity (10 microM genistein) and by the inhibition of cell division (20 microM fluorodeoxyuridine). SCM also increased the uptake of [3H]-choline by retinal cells. The effect was time dependent. The maximum effect was obtained after 48 h and was maintained at a high level until 120 h. Treatment by 10 microM genistein and 15 microM bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM) (an intracellular calcium chelator) completely blocked this effect. However, 20 microM fluorodeoxyuridine did not abolish it. Conditioned medium obtained from glial cells stimulated with SCM (S-GCM) induced an effect on [3H]-choline uptake earlier than that promoted by SCM. Anti-interleukin-2 (IL-2) antibodies blocked the effect of both SCM and S-GCM on [3H]-choline uptake after 48 and 72 h. IL-2 (50 U/ml) elicited the same effect as that observed when the cells were maintained in the presence of SCM. Taken together, our results suggest that IL-2 plays an important role in controlling the survival and differentiation of retinal cells in vitro. PMID- 10810254 TI - Intratympanic and systemic dexamethasone for Meniere's disease. AB - Intratympanic and systemic dexamethasone treatment of Meniere's disease (MD) was evaluated in a prospective study. Seventeen patients (6 men and 11 women) with MD (5 right-sided, 11 left-sided and 1 bilateral) were treated with three 0.2- to 0.4-ml injections of intratympanic dexamethasone hyaluronate (16 mg/ml) during a week and with an initial intramuscular dexamethasone injection of 15 mg. Most of patients were in stage 3, and the mean duration of MD was 5. 3 years. Pure-tone and speech audiometry and the symptom scale of the patients were followed up for 1 year after the treatment. Symptoms of aural fullness, hearing loss, tinnitus and vertigo did not improve significantly. However, sufficient control of vertigo was achieved in 76% of the patients. In conclusion, no definite treatment effect has yet been shown for intratympanic and systemic dexamethasone treatment. Therefore, the clinical use of dexamethasone in MD needs further investigation. PMID- 10810255 TI - Prognostic factors of chronic rhinosinusitis under long-term low-dose macrolide therapy. AB - We studied the immunological and histopathological factors that affect the prognosis of chronic rhinosinusitis under long-term low-dose macrolide therapy. Sixteen patients with chronic rhinosinusitis were given 200 mg clarithromycin or 150 mg roxithromycin orally once a day without other concurrent treatments for 2 3 months. Measurement of the serum IgE level, blood cell count and differential leukocyte count of the peripheral blood, cytological assessment of the nasal smear and computed tomographic (CT) scans of the paranasal sinuses were performed before treatment. The opacity of the sinuses was estimated and scored by the CT images. After treatment, anterior ethmoidal mucosa samples were collected, an infiltrated inflammatory cells, interferon (IFN)-gamma-positive cells and interleukin (IL)-4-positive cells were examined histologically and immunohistochemically. The severity of nasal symptoms was scored before and after treatment, and the improvement rate of the score (symptomatic improvement rate) was calculated. Patients with normal levels of serum IgE (7.10; II, pH range 6.60-6.20; III, pH 5. 47-5.10; IV, pH 5.03-4.60; V, pH 4.76-4.12; VI, pH 4.05 3.82 and VII, pH <3.80) were then tested for their capacity to stimulate cAMP release, androgen aromatization and tissue-type plasminogen activator (tPA) enzyme activity and cytochrome P450 aromatase, tPA and inhibin alpha-subunit mRNA production by rat granulosa cells in culture. cAMP and oestradiol production were determined by RIA, tPA enzyme activity by SDS-PAGE and zymography and all mRNAs by northern blot hybridization analysis and semiquantitative RT-PCR. All isoforms, with the exception of isoform I, stimulated synthesis and release of cAMP, oestrogen and tPA enzyme activity in a dose-dependent manner; the potency of the less acidic isoforms (pH 6. 60-4.60) was greater than that exhibited by the more acidic/sialylated analogs (pH 4.76 to <3.80; potencies II>III>IV>V>VII>VI). A similar trend was observed in terms of cytochrome P450 aromatase and tPA mRNA production. In contrast, when FSH-stimulated production of alpha-inhibin mRNA was analysed, isoforms V-VII were significantly more potent (two- to threefold) than the less acidic/sialylated counterparts (II-IV). In contrast to isoforms II-VII (which behaved as FSH agonists), isoform I (elution pH >7.10) completely blocked P450 aromatase and tPA mRNA expression, without altering that of a constitutively expressed gene (glyceraldehyde-3-phosphate dehydrogenase). These results show for the first time that the naturally occurring human FSH isoforms may exhibit differential or even unique effects at the target cell level. PMID- 10810284 TI - Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture. AB - We tested the hypotheses that the secretion of dimeric inhibin-A from cultured bovine granulosa cells is stimulated by FSH, and that antral cells secrete more inhibin-A than do mural cells. Cells from the antral or mural compartment of follicles were cultured in defined medium in two culture systems, and dimeric inhibin-A was measured by two-site ELISA or by Western immunoblotting. In the first culture system, dimeric inhibin-A secretion declined with time in culture, but was significantly (P<0.05) higher from antral than from mural cells (as was total inhibin-alpha measured by RIA). The secretion of dimeric inhibin-A and inhibin-alpha from antral but not mural cells was responsive to FSH. In the second culture system, dimeric inhibin-A secretion increased with time in culture, and was significantly stimulated by FSH, but FSH responsiveness was dependent on the concentrations of insulin in the culture medium. The major forms of inhibin-A secreted had molecular masses of approximately 58, 62, 103-116 and >116 kDa; the 32 kDa form was barely detectable. These different forms were all stimulated by FSH, but the >116 and 62 kDa forms were most responsive to FSH. We conclude that (i) FSH stimulates dimeric inhibin-A secretion from bovine granulosa cells, (ii) the 62 kDa form of inhibin-A may be more responsive to FSH than the 58 kDa form, and (iii) the spatial differentiation of granulosa cell function within the follicle previously observed for oestradiol secretion was also observed for inhibin-alpha and dimeric inhibin-A secretion. PMID- 10810285 TI - Localization of type 1 17beta-hydroxysteroid dehydrogenase mRNA and protein in syncytiotrophoblasts and invasive cytotrophoblasts in the human term villi. AB - The 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) play a key role in the synthesis of sex steroids. The hallmark of this family of enzymes is the interconversion, through their oxydoreductive reactivity at position C17, of 17 keto- and 17beta-hydroxy-steroids. Because this reaction essentially transforms steroids having low binding activity for the steroid receptor to their more potent 17beta-hydroxysteroids isoforms, it is crucial to the control of the physiological activities of both estrogens and androgens. The human placenta produces large amounts of progesterone and estrogens throughout pregnancy. The placental type 1 17beta-HSD enzyme (E17beta-HSD) catalyzes the reduction of the low activity estrogen, estrone, into the potent estrogen, estradiol. We studied the cell-specific expression of type 1 17beta-HSD in human term placental villous tissue by combining in situ hybridization to localize type 1 17beta-HSD mRNA with immunohistochemistry using an antibody against human placental lactogen, a trophoblast marker. Immunolocalization of E17beta-HSD was also performed. To ascertain whether other steroidogenic enzymes are present in the same cell type, cytochrome P450 cholesterol side-chain cleavage (P450scc), P450 aromatase, and type 1 3beta-hydroxysteroid dehydrogenase (3beta-HSD) were also localized by immunostaining. Our results showed that the syncytium is the major steroidogenic unit of the fetal term villi. In fact, type 1 17beta-HSD mRNA and protein, as well as P450scc, P450 aromatase, and 3beta-HSD immunoreactivities were found in these cells. In addition, our results revealed undoubtedly that extravillous cytotrophoblasts (CTBs), e.g. those from which cell columns of anchoring villous originate, also express the type 1 17beta-HSD gene. However, CTBs lying beneath the syncytial layer, e.g. those from which syncytiotrophoblasts develop, contained barely detectable amounts of type 1 17beta-HSD mRNA as determined by in situ hybridization. These findings, along with those from other laboratories confirm the primordial role of the syncytium in the synthesis of steroids during pregnancy. In addition, our results indicate for the first time that CTBs differentiating along the invasive pathway contain type 1 17beta-HSD mRNA. PMID- 10810286 TI - Expression of inhibin alpha in adrenocortical tumours reflects the hormonal status of the neoplasm. AB - Inhibins are gonadal glycoprotein hormones whose main endocrine function is to inhibit pituitary FSH secretion. In addition to testes and ovaries, other steroid producing organs are sites of inhibin alpha subunit expression. To study the role of inhibins in human adrenal gland, we screened a panel of 150 adrenals (10 normal adrenals, 25 adrenocortical hyperplasias, 65 adrenocortical adenomas, 30 adrenocortical carcinomas and 20 phaeochromocytomas) for inhibin alpha expression. mRNA levels of inhibin alpha subunit were studied in 57 samples and all tissues were stained immunohistochemically with an inhibin alpha subunit specific antibody. Inhibin alpha mRNA was detected in all adrenocortical tissues. Virilizing adenomas possessed a 10-fold higher median inhibin alpha mRNA expression than did normal adrenals. Bilaterally and nodularly hyperplastic adrenals and other than virilizing adrenocortical tumours had their median inhibin alpha mRNA levels close to those of normal adrenals. Immunohistochemically, inhibin alpha subunit was detectable in all normal and hyperplastic adrenals, as well as in 73% of the adrenocortical tumours. However, the percentage of inhibin alpha-positive cells varied greatly in different tumour types. The median percentage of positive cells was 10 in non-functional and Conn's adenomas, 30 in Cushing's adenomas and 75 in virilizing adenomas. In malignant adrenocortical tumours the median percentage of inhibin alpha immunopositive cells was 20 in non-functional carcinomas, 30 in Conn's carcinomas, 65 in Cushing's carcinomas and 75 in virilizing carcinomas. All phaeochromocytomas were negative for inhibin alpha subunit both at the mRNA level and immunohistochemically. Our data show that inhibin alpha subunit is highly expressed in both normal and neoplastic androgen-producing adrenocortical cells, with less expression in cortisol-producing and hardly any in aldosterone producing cells. This suggests a specific role for inhibins in the regulation of adrenal androgen production. We did not find any significant difference in inhibin alpha expression between benign and malignant adrenocortical tumours. Thus inhibin alpha gene does not seem to have a tumour suppressor role in human adrenal cortex. PMID- 10810287 TI - The expression of the IGF system in the bovine uterus throughout the oestrous cycle and early pregnancy. AB - The IGF system is expressed in the uterus during the oestrous cycle and early pregnancy and is likely to play an important role in regulating the development of the embryo and uterus. The IGF peptides (IGF-I and -II) mediate their effects through the type 1 IGF receptor (IGF-1R), while the IGF-binding proteins (IGFBP-1 to -6) modulate their interaction with the receptor. In this study, the expression of the IGF system in the bovine uterus was determined throughout the oestrous cycle and on day 16 of pregnancy. Endometrial biopsy samples were collected from four cows over three cycles such that there were samples for every 2 days from day 0 (oestrus) to day 14 and then every day until day 21. To assess the effect of pregnancy, uterine horn cross-sections were collected on day 16 from 15 pregnant (PREG), five inseminated non-pregnant (INP) and nine uninseminated cyclic controls (CONT). The expression of mRNA for the IGFs, IGF-1R and IGFBP-1 to -5 was determined by in situ hybridisation and the results were quantified by measuring the optical density units from autoradiographs. The main region of IGF-I mRNA expression was the sub-epithelial stroma underlying the luminal epithelium. The expression of IGF-I mRNA was highest at oestrus and lowest during the early and late luteal phases. On day 16, IGF-I mRNA levels were low in all groups, with pregnancy having no effect on the IGF-I mRNA concentrations. The strongest expression of IGF-II mRNA was in the caruncular stroma, with pregnancy having no significant effect in this region. IGF-1R mRNA was also present in the caruncles and was strongly expressed in all epithelial cells both throughout the oestrous cycle and during early pregnancy. The expression of IGFBP-1 mRNA was confined to the luminal epithelium, with the strongest expression seen on day 14 of the cycle. On day 16 the expression of IGFBP-1 mRNA was higher in the PREG group compared with the CONT group. The expression of IGFBP-2 mRNA was localised to the sub-epithelial stroma with more INP than PREG cows showing detectable levels of IGFBP-2. The strongest expression of IGFBP-3 mRNA was in the caruncular stroma; expression in the endometrial stroma was similarly decreased during early pregnancy. IGFBP-5 mRNA was mainly expressed in the inner ring of myometrium and was not affected by pregnancy on day 16. In conclusion, these results show that many components of the uterine IGF system are differentially regulated during the oestrous cycle and early pregnancy and suggest that modulation of the IGF system may influence uterine activity during this period. PMID- 10810288 TI - Insulin-like growth factor II in human fetal pancreas and its co-localization with the major islet hormones: comparison with adult pancreas. AB - Insulin-like growth factor II (IGF-II) appears to play an important role during fetal life in cell growth and differentiation in several organs, including the pancreas. In the present study we investigated the cellular localization of IGF II in human fetal pancreas at 16, 18 and 22 embryonic weeks and compared it with adult pancreas. Single and double immunofluorescence methods were used to study co-localization of IGF-II with the four major islet hormones - insulin, glucagon, somatostatin, pancreatic polypeptide - and with islet amyloid polypeptide (IAPP). Distinct IGF-II immunoreactive (IR) cells were found in the endocrine, but not in the exocrine, pancreas. The intensity of IGF-II immunoreactivity was more pronounced in the fetal than in the adult pancreas. In fetal pancreas IGF-II immunoreactivity was observed in virtually all insulin-IR cells and in subsets of the glucagon, somatostatin and IAPP cells. In the adult pancreas, IGF-II immunoreactivity was found in insulin/IAPP cells only. Our results suggest a broader effect of IGF-II in fetal endocrine pancreatic cells than in the adult. PMID- 10810289 TI - Binding characteristics of pro-insulin-like growth factor-II from cancer patients: binary and ternary complex formation with IGF binding proteins-1 to -6. AB - Many tumours secrete IGF-II in incompletely processed precursor forms. The ability of these pro-IGF-II forms to complex with the six IGF binding proteins (IGFBPs) is poorly understood. In this study, pro-IGF-II has been extracted from the serum and tumour tissue of two patients with non-islet cell tumour hypoglycaemia. These samples were used to study binary complex formation with IGFBPs-1 to -6 using competitive IGF-II binding assays and ternary complex formation with IGFBP-3 and IGFBP-5. In each case, IGFBPs-1 to -6 showed little difference in their ability to form binary complexes with recombinant IGF-II or tumour-derived pro-IGF-II forms, when the preparations were standardised according to IGF-II immunoreactivity. As previously described, ternary complex formation by acid-labile subunit (ALS) with IGFBP-3 and pro-IGF-II was greatly decreased compared with complex formation with mature IGF-II. In contrast, ALS bound similarly to IGFBP-5 in the presence of pro-IGF-II and mature IGF-II. These studies suggest that pro-IGF-II preferentially forms binary complexes with IGFBPs, and ternary complexes with IGFBP-5, rather than ternary complexes with IGFBP-3 as seen predominantly in normal serum. This may increase the tissue availability of serum pro-IGF-II, allowing its insulin-like potential to be realised. PMID- 10810290 TI - Effect of IGF-I on serine metabolism in fetal sheep. AB - Acute infusion of IGF-I to the fetus has been shown to inhibit amino acid oxidation and appears to increase fetoplacental amino acid uptake. This study was designed to investigate further the effects of IGF-I on fetal amino acid metabolism. Radiolabeled serine was used to test the hypothesis that fetal IGF-I infusion enhances serine uptake into the fetus and/or placenta and inhibits serine oxidation. Eight fetal sheep were studied at 127 days of gestation before and during a 4-h infusion of IGF-I (50 microg/h per kg). During the infusion there was no change in uptake of serine or its oxidation by fetus or placenta. However, both uptake and oxidation of serine and glycine decreased in the fetal carcass. There was also a decrease in fetal blood serine and glycine concentrations which could indicate a decrease in protein breakdown, although reduced amino acid synthesis cannot be excluded. Thus IGF-I appeared to influence the distribution of these amino acids as oxidative substrates between different fetal tissues. In addition, fetal IGF-I infusion increased the conversion of serine to glycine which is likely to have increased the availability of one carbon groups for biosynthesis. Our data provide further evidence that IGF-I plays a role in the regulation of fetoplacental amino acid metabolism. PMID- 10810291 TI - Measurement of the acid-labile subunit of the insulin-like growth factor binding protein complex in human serum: a comparison of four immunoassays. AB - The acid-labile subunit (ALS) of the high molecular weight insulin-like growth factor binding protein complex is a liver-derived glycoprotein which is regulated by growth hormone and serves as a serum marker of growth hormone action. We have compared the measurement of ALS by four immunoassay methods (two RIAs, two ELISAs) utilizing various polyclonal and monoclonal antibodies raised against natural or recombinant human ALS, or synthetic ALS peptides. Despite the variety of methodologies and reagents, results obtained by the four methods were highly correlated for 125 sera from various patient groups, and when compared for individual groups of sera from healthy children and adults, growth hormone deficient children and adults, and subjects with acromegaly. Some weaker correlations among methods were seen when measuring ALS levels in groups of sera from pregnant subjects and subjects with chronic renal failure. An assay using antibodies raised against recombinant ALS yielded lower apparent values than the other methods in patient sera, the discrepancy probably being attributable to a difference in standardization. We conclude that a variety of assay formats and reagents can yield serum ALS values of potential clinical utility. PMID- 10810292 TI - Role of glucose in chronic desensitization of isolated rat islets and mouse insulinoma (betaTC-3) cells to glucose-dependent insulinotropic polypeptide. AB - It is well documented that the release of insulin from isolated perifused islets attenuates over time, despite a continued glucose stimulation. In the current study we have shown that potentiation of insulin release by the intestinal hormone glucose-dependent insulinotropic polypeptide (GIP) is also attenuated after its continuous application. In less than 20 h of maintained stimulus with either hyperglycaemia (11.0 mM glucose) or GIP (10 nM) under hyperglycaemic conditions, insulin release returned to basal values. This was not due to loss of islet viability or reduction in the releasable pool of insulin granules, as 1 mM isobutylmethylxanthine was able to stimulate equivalent insulin release under both conditions. Further examination of chronic GIP desensitization was examined in cultured mouse insulinoma (betaTC-3) cells. GIP-stimulated cAMP production was not greatly affected by the prevailing glucose conditions, suggesting that the glucose dependence of GIP-stimulated insulin release occurs distally to the increase in intracellular cAMP in betaTC-3 cells. The GIP-stimulated cAMP response curve after desensitization was of similar magnitude at all glucose concentrations, but GIP pretreatment did not affect forskolin-stimulated cAMP production. Desensitization of the cAMP response in betaTC-3 cells was shown not to involve induction of dipeptidyl peptidase IV or pertussis toxin-sensitive G proteins, activation of protein kinase C or protein kinase A, or modulation of phosphodiesterase activity. Homologous desensitization of the insulin potentiating activity of GIP was found to affect both GIP-stimulated and forskolin-stimulated insulin release, indicating desensitization of distal steps in the stimulus-exocytosis cascade. PMID- 10810293 TI - Nitric oxide, islet acid glucan-1,4-alpha-glucosidase activity and nutrient stimulated insulin secretion. AB - The mechanism of nutrient-evoked insulin release is clearly complex. One part of that mechanism is postulated to be the activation of the glycogenolytic enzyme acid glucan-1,4-alpha-glucosidase. As nitric oxide (NO) has been found to be a potent inhibitor of glucose-stimulated insulin secretion, we have now investigated a possible influence of exogenous NO and inhibition of endogenous NO production on islet acid glucan-1,4-alpha-glucosidase activity in relation to insulin release stimulated by glucose and l-arginine. In isolated islets, NO derived from the intracellular NO donor hydroxylamine inhibited the activation of acid glucan-1, 4-alpha-glucosidase and its isoform acid alpha-glucosidase in parallel with inhibition of glucose-stimulated insulin release. In comparison, other lysosomal enzymes were largely unaffected. Similarly, the spontaneous NO donor sodium nitroprusside, as well as NO gas, when added to islet homogenates, suppressed the activities of these acid alpha-glucosidehydrolases and, to a lesser extent, the activities of other lysosomal enzymes. Finally, in the presence of the NO synthase inhibitor N(G)-nitro-l-arginine methyl ester, insulin release from isolated islets stimulated by glucose or l-arginine was markedly potentiated in parallel with an accompanying increase in the activities of acid glucan-1,4-alpha-glucosidase and acid alpha-glucosidase. Other lysosomal enzymes and neutral alpha-glucosidase were not influenced. We propose that an important inhibitory effect of NO on the insulin secretory processes stimulated by glucose and l-arginine is exerted via inactivation of islet acid glucan-1,4-alpha glucosidase, a putative key enzyme in nutrient-stimulated insulin release. PMID- 10810294 TI - Stimulation of endogenous GH and interleukin-6 receptors selectively activates different Jaks and Stats, with a Stat5 specific synergistic effect of dexamethasone. AB - The interaction of GH, interleukin (IL)-6 and glucocorticoids is likely to be important in regulating the GH-insulin-like growth factor (IGF)-I axis. The signalling cascades activated by GH and IL-6 appear to be very similar, as demonstrated by studies using overexpression of the receptor and other components of the Jak-Stat and mitogen-activated protein (MAP) kinase pathways. Here we show that the human embryonic kidney cell line 293 (HEK293) expresses GH and IL-6 receptors endogenously. To determine which specific pathways might be activated by the two cytokines, at physiological levels of all components, we studied GH and IL-6 mediated signal transduction both under basal conditions and in the presence of overexpressed receptors and Stat proteins. Our results suggest a receptor specificity of Jak2 for GH receptors, and Jak1 for IL-6 receptors. Stat activation in response to GH and IL-6 was determined by reporter gene induction. Both GH and IL-6 were able to induce the reporter gene containing the Stat5 responsive element (LHRE) but the IL-6 response appeared to be mediated mainly through Stat3 activation. In contrast, the reporter gene containing the Stat3 responsive element (SIE) was IL-6 specific. The levels of gene induction by GH and IL-6 were not altered by the co-stimulation with GH and IL-6, suggesting that there is little cross-talk at the Jak-Stat activation level between the two cytokines. Neither GH nor IL-6 activated the MAP-kinase responsive serum response element (SRE), unless GH receptors or gp130 were overexpressed. Transfection of Stat3 or Stat5 expression vectors enhanced the response to GH and IL-6. Stimulation with dexamethasone synergistically enhanced GH activation of the LHRE reporter gene but had no effect on the IL-6 activation of the same reporter or on the SIE reporter gene. Thus, our studies suggest that while each cytokine, GH and IL-6, may activate various members of the Jak-Stat pathway in overexpression studies, specific activation of Stat3 by IL-6 and of Jak2 and Stat5 by GH can be observed in HEK293 cells and that in this system the synergistic effect of dexamethasone appears specific for Stat5. PMID- 10810295 TI - Regulation of human GH receptor gene transcription by 20 and 22 kDa GH in a human hepatoma cell line. AB - The human GH gene is 1.7 kilobase pairs (kb) in length and is composed of five exons and four introns. This gene is expressed in the pituitary gland and encodes a 22 kDa protein. In addition to this predominant (75%) form, 5-10% of pituitary GH is present as a 20 kDa protein that has an amino acid (aa) sequence identical to the 22 kDa form except for a 15 aa internal deletion of residues 32-46 as a result of an alternative splicing event. Because it has been reported that non-22 kDa GH isoforms might be partly responsible for short stature and growth retardation in children, the aim of this study was to compare the impact of both 22 kDa and 20 kDa GH on GH receptor gene (GH receptor/GH binding protein (GHR/GHBP)) expression. Various concentrations of 20 kDa and 22 kDa GH (0, 2, 5, 12.5, 25, 50 and 150 ng/ml) were added to human hepatoma (HuH7) cells cultured in serum-free hormonally defined medium for 0, 1 and 2 h. Thereafter GHR/GHBP mRNA expression was measured by quantitative PCR. Addition of either 20 kDa or 22 kDa GH, at low or normal physiological concentrations (0, 2, 5, 12.5, 25 or 50 ng/ml) induced a dose-dependent increase in GHR/GHBP expression. However, a supraphysiological concentration of 20 kDa GH (150 ng/ml) resulted in a significantly lower (P<0.05) downregulation of GHR/GHBP gene transcription compared with the downregulation achieved by this concentration of 22 kDa GH. This difference might be explained by a decreased ability to form a 1 : 1 complex with GHR and/or GHBP, which normally occurs at high concentrations of GH. Nuclear run-on experiments and GHBP determinations confirmed the changes in GHR/GHBP mRNA levels. In conclusion, we report that both 20 kDa and 22 kDa GH, in low and normal physiological concentrations, have the same effect on regulation of GHR/GHBP gene transcription in a human hepatoma cell line. At a supraphysiological concentration of 150 ng/ml, however, 20 kDa GH has a less self inhibitory effect than the 22 kDa form. PMID- 10810296 TI - Transgenic mice expressing bovine GH develop arthritic disorder and self antibodies. AB - We observed disability of movement in 6-month-old transgenic mice expressing the fusion gene coding for the bovine GH (bGH) under the transcriptional control of phosphoenolpyruvate carboxykinase promoter (PEPCK-bGH). Histological study of the knee joint showed altered synovial and tibial articular cartilage tissues. In the cartilage the following observations were made: (i) generalized loss of the normal zonal structure and presence of clefts, and (ii) profound alterations in chondrocyte growth/differentiation processes consistent with hypertrophy. The synovial tissue showed a reduced number of adipocytes, and a significant thickening of synovial lining tissue and pannus. These findings indicate that transgenic mice suffer damage to diarthritic joints with osteoarthritic appearance. As changes in synovial membrane in osteoarthritis are almost indistinguishable from those seen in inflammatory arthritis, we determined the potential correlation with an immunological disorder. Serological determination of self-antibodies measured as a function of age and sex showed anti-nuclear, anti-single-stranded DNA, anti-double-stranded DNA and anti-70K antibodies, and an altered immunoglobulin typing. These results suggest that transgenic mice expressing bGH develop an arthritic process which is correlated with an immune disorder. The results also indicate that these mice are a suitable animal model to study the specific role of GH-driven processes in immune cells and arthritis. PMID- 10810297 TI - Optimisation of growth hormone production by muscle cells using plasmid DNA. AB - The production of peptide hormones by skeletal muscle tissue is a promising area of gene therapy. Skeletal muscle myogenesis can be induced in vitro, resulting in the fusion of mononucleate myoblasts to form multinucleate myotubes, and delivery vectors are first tested in vitro. C2C12 myoblasts transfected with pcDNA3-GH, which used the human cytomegalovirus (CMV) promoter, secreted immunoreactive GH with comparable biological activity to pituitary GH. Mouse myeloid leukaemia cells, which express the mouse GH receptor were used for the bioassay, and activation of these cells by GH was measured by a colorimetric microculture tetrazolium assay. Cells were incubated with a tetrazolium salt (MTS) and an intermediate electron acceptor (phenazine methosulphate, PMS), and formazan production was measured as optical density (O.D.) at 490 nm. The efficiencies of several plasmid expression vectors were compared in differentiated and non differentiated muscle cells, as a function of bioactive GH secreted by the transfected cells. Ten-day differentiated C2C12 myotubes transfected with pcDNA3E GH, which used the CMV promoter and a rat myosin light chain enhancer element, secreted significantly more biologically active GH than myotubes transfected with pcDNA3-GH (0.82 O.D. units+/-0.06 vs 0.57+/-0.05 respectively, P<0.001). This was consistent with reduced CMV promoter activity in myotubes. Myoblasts transfected with pcDNA3-GH secreted more bioactive GH than 10-day transfected myotubes (1.1+/ 0. 1 vs 0.77+/-0.07 respectively). However, the responses were indistinguishable (both 1.0+/-0.09) if both the myotubes and myoblasts had been transfected with pcDNA3E-GH. Substitution of the vector pMHLC-GH, which used a muscle-specific truncated rabbit myosin heavy chain promoter, and the myosin enhancer resulted in a marked decrease in the responses to the conditioned medium from fused myotubes compared with the vectors pcDNA3-GH and pcDNA3E-GH (0. 24+/-0.02 vs 0.57+/-0.05 vs 0.82+/-0.06 respectively). We concluded that the combination of CMV promoter and myosin light chain enhancer in pcDNA3E-GH had the greatest expression efficiency of the several plasmid vectors which we investigated. PMID- 10810298 TI - Testosterone modulates growth hormone secretion at the hypothalamic but not at the hypophyseal level in the adult male rhesus monkey. AB - We investigated a possible modulation of growth hormone (GH) secretion by testosterone by measuring the growth hormone releasing hormone (GHRH)-stimulated and N-methyl-d,l-aspartic acid (NMA)-induced GH secretion in adult rhesus monkeys. Intact, orchidectomized and testosterone-substituted (testosterone enanthate 125 mg/week, i.m. for 5 weeks) orchidectomized monkeys (n=5) were used in the study. GHRH (25 microg/kg body weight) or NMA (15 mg/kg body weight) was infused through a Teflon cannula implanted in the saphenous vein. Sequential blood samples were collected 30-60 min before and 60 min after the injection of the neurohormone or the drug at 10-20-min intervals. All bleedings were carried out under ketamine hydrochloride anaesthesia (initial dose 5 mg/kg body weight i.m., followed by 2.5 mg/kg at 30-min intervals). The plasma concentrations of GH, testosterone and oestradiol (E(2)) were determined by using specific assay systems. Administration of GHRH elicited a significant increase in GH secretion in all three groups of animals. There was no significant difference in the responsiveness of pituitary somatotrophs to exogenous GHRH challenges between intact and orchidectomized monkeys and testosterone replacement in orchidectomized animals did not significantly alter the GHRH-induced GH response. The responsiveness of hypothalamic GHRH neurones apparently did undergo a qualitative change after orchidectomy, as GH response to NMA was less in orchidectomized animals than in intact monkeys. The responsiveness of GHRH neurones to exogenous NMA was restored and even potentiated when orchidectomized monkeys were treated with testosterone. Taken together, these findings suggest that testosterone does not affect the sensitivity of the pituitary somatotrophs to GHRH but stimulates the secretion of GH by modulation of the NMDA drive to GHRH neurones. PMID- 10810299 TI - Neonatal exposure to estrogen differentially alters estrogen receptor alpha and beta mRNA expression in rat testis during postnatal development. AB - The biological actions of estrogens on target cells are mediated by two nuclear receptors: the estrogen receptor (ER) alpha and the recently characterized ER beta. In the male rat, the physiological role of estrogens involves multiple actions, from masculinization of brain areas related to reproductive function and sexual behavior to regulation of testicular development and function. Paradoxically, however, administration of high doses of estrogen during the critical period of neonatal differentiation results in an array of defects in the reproductive axis that permanently disrupt male fertility. The focus of this study was to characterize the effects and mechanism(s) of action of neonatal estrogenization on the pattern of testicular ER alpha and beta gene expression during postnatal development. To this end, groups of male rats were treated at day 1 of age with estradiol benzoate (500 microg/rat), and testicular ER alpha and ER beta mRNA levels were assayed by semi-quantitative RT-PCR from the neonatal period until puberty (days 1-45 of age). Furthermore, the expression of androgen receptor (AR) mRNA was evaluated, given the partially overlapping pattern of tissue distribution of ER alpha, ER beta and AR messages in the developing rat testis. In addition, potential mechanisms for neonatal estrogen action were explored. Thus, to discriminate between direct effects and indirect actions through estrogen-induced suppression of serum gonadotropins, the effects of neonatal estrogenization were compared with those induced by blockade of gonadotropin secretion with a potent LHRH antagonist in the neonatal period. Our results indicate that neonatal exposure to estrogen differentially alters testicular expression of alpha and beta ER messages: ER alpha mRNA levels, as well as those of AR, were significantly decreased, whereas relative and total expression levels of ER beta mRNA increased during postnatal/prepubertal development after neonatal estrogen exposure, a phenomenon that was not mimicked by LHRH antagonist treatment. It is concluded that the effect of estrogen on the expression levels of ER alpha and beta mRNAs probably involves a direct action on the developing testis, and cannot be attributed to estrogen-induced suppression of gonadotropin secretion during the neonatal period. PMID- 10810300 TI - Localization of oestrogen receptor alpha, oestrogen receptor beta and androgen receptors in the rat reproductive organs. AB - There is now evidence that oestrogens and androgens can influence male and female reproductive systems. In order to accurately identify the sites of action of oestrogens and androgens, we have proceeded to the histological localization of the two oestrogen receptor (ER) subtypes, ERalpha and ERbeta, and the androgen receptor (AR) in the reproductive tissues of adult rats of both sexes. AR was detected by immunocytochemistry, while ERalpha and ERbeta were localized by both immunocytochemistry and in situ hybridization. In the pituitary gland of animals of both sexes, ERalpha was found in the majority of nuclei of secretory cells in the anterior pituitary. The intermediate and posterior lobes did not show any staining. ERbeta was not found to be expressed in any of the pituitary lobes. Using AR antibodies, nuclear staining was detected in about 50% of secretory cells of the anterior lobe, the intermediate and posterior lobes being completely unstained. In the testis, ERalpha was localized in nuclei of Leydig cells as well as in round spermatocytes and spermatids, while ERbeta could only be detected in Sertoli cell nuclei. AR immunoreactivity was found in nuclei of Sertoli, peritubular myoid and Leydig cells. In the prostate, ERbeta was observed in epithelial cells of tubulo-alveoli, while the stroma was unlabelled. ERalpha was not found to be expressed in any prostate cells. In the prostate, AR was detected in nuclei of epithelial, stromal and endothelial cells. In seminal vesicles, staining of ERalpha was found in nuclei of epithelial and stromal cells. Similar findings were observed using AR antibodies. While ERbeta mRNA could not be detected by in situ hybridization, weak staining for ERbeta was localized in epithelial cells of seminal vesicles. In the ovary, both ERalpha and ERbeta were found to be expressed. ERbeta mRNA was found in granulosa cells of growing follicles, while ERalpha was present in theca cells, interstitial gland cells and germinal epithelium. AR immunoreactivity was detected in granulosa cell nuclei in growing follicles and also in scattered interstitial cells. In the oviduct and uterus, ERalpha was observed in nuclei of epithelial cells as well as of stromal and muscle cells. Similarly, AR immunoreactivity was present in nuclei of epithelial cells, stromal and muscle cells in both the oviduct and uterus. ERbeta was not detected in the oviduct and uterus. The present findings indicate a cell specific localization of ERalpha, ERbeta and AR in reproductive tissues in rats of both sexes. By establishing the precise sites of action of oestrogens and androgens they contribute to a better understanding of the respective role of these steroids in reproduction function. PMID- 10810301 TI - Regulation of estrogen receptor-alpha gene expression by epidermal growth factor. AB - The role of epidermal growth factor (EGF) in the regulation of estrogen receptor alpha (ER-alpha) gene expression in the human breast cancer cell line MCF-7 was investigated. Treatment of cells with 0.4 ng/ml EGF resulted in an approximately 60% decrease in ER-alpha protein concentration by 6 h and the amount of receptor remained suppressed for 24 h. Ligand binding assays demonstrated that the decrease in ER-alpha protein corresponded to a similar decrease (approximately 50%) in estradiol binding sites. Although EGF treatment resulted in a decrease in the number of binding sites, it had no effect on the binding affinity of ER alpha. The dissociation constant of the estradiol-ER-alpha complex in the presence or absence of EGF was the same (K(d)=2.3x10(-)(10) M in control cells versus K(d)=1.98x10(-)(10) M in EGF-treated cells). The decrease in ER-alpha protein concentration paralleled a decrease in the steady-state amount of ER alpha mRNA. By 9 h there was an approximately 60% decrease in ER-alpha mRNA. The amount of ER-alpha mRNA remained suppressed for 48 h. Transcription run-on experiments demonstrated that there was a decrease of approximately 70% in ER alpha gene transcription upon EGF treatment, suggesting that the mechanism by which EGF regulates ER-alpha gene expression is transcriptional. In addition to regulating the amount of ER-alpha, EGF affected the activity of the receptor. At high concentrations, EGF induced progesterone receptor. Estradiol and high concentrations of EGF had an additive effect on progesterone receptor. In contrast to high concentrations, low concentrations of EGF had no effect on progesterone receptor and blocked estradiol induction. The effects of EGF on ER alpha expression were inhibited by tyrophostins and wortmannin, suggesting that the effects of the growth factor are mediated by the EGF receptor and protein kinase B. When the cells were placed in serum-free medium and then treated with EGF, there was no effect on ER-alpha protein concentration or activity. However, increasing concentrations of serum restored the effects of EGF on ER-alpha, suggesting that an additional serum factor was required for the EGF-mediated effect on the decrease in ER-alpha protein concentration. PMID- 10810302 TI - Overexpression of thyroid hormone receptor beta1 is associated with thyrotropin receptor gene expression and proliferation in a human thyroid carcinoma cell line. AB - To correlate the differentiation phenotype of two human thyroid cancer cell lines with their expression of various molecular markers, we analyzed the mRNA levels of four thyroid-specific genes, including thyrotropin receptor (TSHR), thyroglobulin (Tg), thyroid transcription factor-1 (TTF-1), and paired-box containing transcription factor-8 (PAX-8) genes. The results showed a differentiation-status-related pattern in which a well-differentiated cell line (WRO) expressed all the four genes, in contrast to an anaplastic cell line (ARO) that expressed TTF-1 and reduced levels of TSHR, but no Tg or PAX-8 genes. Furthermore, to verify the finding of concomitant loss of beta subtype thyroid hormone receptor (TRbeta) and TSHR gene expression in neoplastic thyroid tumors (Bronnegard et al. 1994), we examined the expression levels of TRbeta1 gene in these cell lines. Whereas the WRO cells produced an abundant amount of TRbeta1 protein detectable by immunoprecipitation, the ARO cells produced none. This new observation prompted us to investigate whether overexpression of TRbeta1 protein in ARO cells might produce changes in the differentiation phenotypes. We found that the level of expression of the TSHR gene and the proliferative index of ARO cells were significantly upregulated in the cells stably transfected with wild type TRbeta1. These findings suggest that TRbeta1 protein overexpression can affect the differentiation phenotypes and induce more efficient cell proliferation of the anaplastic ARO cells. PMID- 10810303 TI - Mechanism of liver-selective thyromimetic activity of SK&F L-94901: evidence for the presence of a cell-type-specific nuclear iodothyronine transport process. AB - The thyromimetic compound SK&F L-94901 shows more potent thyromimetic activity in the liver than in the pituitary gland or heart when administered to rats. The mechanisms of liver-selectivity of SK&F L-94901 were examined using cultured rat hepatoma cells (dRLH-84) and rat pituitary tumor cells (GH3), both of which showed saturable cellular uptake of tri-iodothyronine (T(3)). When isolated nuclei with partial disruption of the outer nuclear membrane were used, SK&F L 94901 competed for [(125)I]T(3) binding to nuclear receptors almost equally in dRLH-84 and GH3 cells. SK&F L-94901 also did not discriminate thyroid hormone receptors (TR) alpha1 and beta1 in terms of binding affinity and activation of the thyroid hormone responsive element. In intact cells, however, SK&F L-94901 was a more potent inhibitor of nuclear [(125)I]T(3) binding in dRLH-84 cells than in GH3 cells at an early phase of the nuclear uptake process and after binding equilibrium. These data suggest that SK&F L-94901 is more effectively transported to nuclear TRs in hepatic cells than in pituitary cells and therefore shows liver selective thyromimetic activity. In conclusion, SK&F L-94901 discriminates hepatic cells and pituitary cells at the nuclear transport process. The cellular transporters responsible for this discrimination were not evident. PMID- 10810304 TI - Localisation and temporal changes in prostaglandin G/H synthase-1 and -2 content in ovine intrauterine tissues in relation to glucocorticoid-induced and spontaneous labour. AB - Parturition in the ewe is preceded by an increase in the synthesis of prostaglandins (PGs) by gestational tissues. To establish the uterine source of these PGs, placental cotyledons, fetal membranes and maternal uterine tissues were collected from ewes (n=6) at spontaneous parturition. Solubilised tissue extracts were prepared and analysed by Western blots using polyclonal antibodies to PG G/H synthase-1 and -2 (PGHS-1 and PGHS-2). PGHS-1 was expressed by all intrauterine tissues at term labour. Densitometric analysis of Western blot autoradiographs showed that the fetal membranes and maternal cervix contained the largest amounts of PGHS-1. PGHS-1 enzyme content of ovine amnion was significantly greater than that of either chorion or allantois (P<0.05). PGHS-1 protein content of myometrial, endometrial and cotyledonary tissue extracts was minimal. Formation of the PGHS-2 isozyme was confined to placental tissue at term labour. PGHS-2 protein levels in sheep placenta were significantly higher than those of PGHS-1 in all intrauterine tissues examined. This result supports the hypothesis that PGHS-2 is a major contributor to PG formation at term labour. To elucidate the developmental changes in PGHS-1 and PGHS-2 relative to labour onset, an experimental paradigm of glucocorticoid-induced delivery was used. Previous characterisation and validation of this labour model demonstrated that direct, transabdominal, intrafetal injection of the synthetic glucocorticoid betamethasone (5.7 mg in 1 ml aqueous vehicle) on day 131 of gestation induced labour onset in 56.6+/-0.8 h (mean+/-s.e.m.). As the latent period to induced labour was known, the time course of enzyme formation could be ascertained. Sheep (n=20) were killed by barbiturate injection at various time intervals post injection (0, 14, 28, 42 and 56 h). Tissue extracts collected at post-mortem examination were prepared and analysed by Western blots. PGHS-2 was induced in ovine cotyledon in a time-dependent fashion following glucocorticoid injection (P<0.05). There was a 12-fold increase in abundance between the time of betamethasone administration (0 h) and established labour (56 h). The PGHS-2 isozyme was not detected in any of the other tissues examined. In contrast, formation of the PGHS-1 isozyme did not change in relation to induced-labour in any of the intrauterine tissues. This finding is consistent with constitutive formation of PGHS-1. Previous studies have demonstrated a rise in PG production in association with glucocorticoid-induced labour and spontaneous delivery. The results of the present study indicate that this rise in PG production is due to increased formation of the PGHS-2 isozyme in ovine cotyledon. PGHS-2 appears to be induced by exogenous glucocorticoid administration and/or the mechanisms controlling ovine parturition. The role of PG formation by the fetal membranes is yet to be elucidated. PMID- 10810305 TI - Anastomosis of the ovarian vein to the hepatic portal vein in sheep induces ovarian hyperstimulation associated with increased LH pulsatility, but only in the absence of the contralateral ovary. AB - In this study, two experiments were performed, the first of which examined the ovarian response in ewes that were subject to unilateral ovariectomy (ULO) at different intervals (0-14 days) after surgical anastomosis (AN) of the ovarian vein to the mesenteric vein (n=7 ewes), or sham operation (SO; n=4 ewes). Hypertrophy and development of multiple follicular and luteal structures on AN ovaries were observed after ULO, while SO ovaries remained of normal size and appearance after ULO. The second experiment involving 11 ewes (five AN; six SO) aimed to clarify the mechanism by which AN following ULO-induced ovarian hypertrophy and increased follicle development. The results confirmed that there were more large (>5 mm) follicles on AN compared with SO ovaries; however, their rate of atresia was similar. Oestradiol and progesterone concentrations in follicular fluid of class 1 follicles (5-9 mm) were higher in AN ovaries than those in control follicles of the same size collected in the late follicular phase of an induced oestrous cycle. In AN ewes, intrafollicular progesterone concentrations increased while follicular aromatase activity and intrafollicular oestradiol, inhibin A, follistatin and activin A concentrations all decreased as follicle size increased. Oestradiol and progesterone concentrations were substantially higher in ovarian venous blood than in hepatic venous blood, both in AN and SO ewes, whereas inhibin A levels were not significantly modified by passage through the liver in either group. Mean plasma LH concentration, and LH pulse frequency and amplitude increased markedly after AN but were not affected by SO. Plasma FSH showed only a small transient increase after AN, presumably due to the maintenance of inhibin feedback. Injection of prostaglandin F(2)(alpha) 4 days later did not further modify LH or FSH secretion in either group. Full ovariectomy (FO) 9-14 days after AN or SO increased LH secretion markedly in SO ewes but to a lesser degree in AN ewes; FO induced a large and rapid increase in FSH levels in both groups. In conclusion, AN of the ovary to the liver via the mesenteric vein provides a useful model for studying the feedback between the ovary and the hypothalamo-pituitary system and the mechanisms controlling follicle development. The present results indicate that the pattern of LH secretion is an important factor controlling the terminal phase of follicle development in the ewe. PMID- 10810306 TI - Prostatic secretory protein (PSP94) expression in human female reproductive tissues, breast and in endometrial cancer cell lines. AB - PSP94 (beta microseminoprotein, beta MSP) is one of the three major proteins secreted by the normal human prostate gland. Using reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blotting, PSP94 transcripts were shown in human endometrium, myometrium, ovary, breast, placenta and in the human endometrial cancer cell lines KLE and AN3 CA. Primers used in these studies were specific for human prostate PSP94, and were derived from its flanking non-coding regions. The results were confirmed by sequence analysis of two independently derived clones from normal human breast tissues and the other two from KLE cells respectively. The sequences were identical with the coding sequence of human prostate PSP94 cDNA. Using RNA from the endometrial tissues, two different transcripts of approximately 487 bp, equivalent to prostate PSP94 and approximately 381 bp, corresponding to prostate PSP57, its alternately spliced form, were amplified by RT-PCR. Human ovary, breast, placenta and endometrial cancer cell lines (KLE, AN3 CA), however, showed only the full length, approximately 487 bp, PSP94 transcript. We further demonstrated by in situ hybridization that PSP94 mRNA is expressed specifically in the glandular epithelial cells, and not in the stroma of both the human endometrial and breast tissues. Further, using image analysis of in situ hybridization data, the levels of PSP94 mRNA in the cycling endometrial tissues and in breast confirmed the differential levels of expression in the cycling endometrium (P<0.005). This study distinctly demonstrated significant expression of PSP94 mRNA in human uterine, breast and other female reproductive tissues as well in the endometrial cancer cell lines, suggesting that it may have a role in these tissues as a local autocrine paracrine factor. PMID- 10810307 TI - A panel of monoclonal antibodies to ovine placental lactogen. AB - A panel of 11 rat monoclonal antibodies (mAbs) has been raised to ovine placental lactogen (PL). By competitive enzyme-linked immunoabsorbent assay (ELISA), confirmed by two-site ELISA, the antibodies were shown to recognize six antigenic determinants on the ovine PL molecule, two of which overlap. One antigenic determinant (designated 1) was shared by other members of the prolactin/growth hormone (GH)/PL family in ruminants, humans and rodents. The binding of (125)I labelled ovine PL to crude receptor preparations from sheep liver (somatotrophic) or rabbit mammary gland (lactogenic) was inhibited by mAbs recognizing antigenic determinants 2-6. Both types of receptor preparation were affected similarly. In the local in vivo pigeon crop sac assay, mAbs directed against determinants 3 and 6 enhanced the biological activity of ovine PL. PMID- 10810308 TI - Placental lactogen-I gene activation in differentiating trophoblast cells: extrinsic and intrinsic regulation involving mitogen-activated protein kinase signaling pathways. AB - Trophoblast giant cells are one of the primary endocrine cell types of the rodent placenta. Placental lactogen-I (PL-I) is the initial prolactin (PRL) family member expressed as trophoblast giant cells differentiate. In this report, we use the Rcho-1 trophoblast cell line as a model for studying the regulation of PL-I gene expression during trophoblast giant cell differentiation. Evidence is provided for trophoblast cell expression of epidermal growth factor receptor (EGFR), ErbB2, fibroblast growth factor receptor 1 (FGFR1), transforming growth factor-alpha, and heparin-binding EGF. EGF and FGF-2 stimulated PL-I mRNA and protein accumulation and PL-I promoter activity in a concentration-dependent manner. These latter growth factor actions on PL-I promoter activities were specifically inhibited by cotransfection with dominant negative constructs for EGFR and FGFRs respectively. Utilization of the mitogen-activated protein kinase (MAPK) pathway by EGF and FGF-2 in trophoblast cells was demonstrated by growth factor stimulation of a Gal4 DNA binding/Elk1 transactivational domain fusion construct, and more specifically by activation of extracellular signal regulated kinase and p38 MAPK. PL-I gene activation was also sensitive to disruption of MAPK and activation protein-1 (AP-1) signaling pathways. In conclusion, autocrine/paracrine pathways involving EGFR and FGFR1, MAPK and AP-1 are shown to participate in the regulation of the PL-I gene in differentiating trophoblast cells. PMID- 10810309 TI - The distribution of stanniocalcin 1 protein in fetal mouse tissues suggests a role in bone and muscle development. AB - We previously isolated a mammalian gene STC1 that encodes a glycoprotein related to stanniocalcin (STC), a fish hormone that plays a major role in calcium homeostasis. However, the mammalian STC1 gene is expressed in a variety of adult tissues in contrast to fish where STC is expressed only in one unique gland, the corpuscles of Stannius. This suggested that STC1 may have wider autocrine/paracrine functions in mammals. In the present study, using immunocytochemistry, we showed that STC1 protein is localized in the developing bone and muscle of the mouse fetus. During endochondral bone formation, STC1 is found principally in prechondrocytes and prehypertrophic chondrocytes. During intramembranous bone formation STC1 is present in the mesenchyme that is about to undergo ossification. STC1 is also found in the myocardiocytes of the developing heart and at all stages of differentiation from myoblasts to myotube formation in developing skeletal muscle. The specific localization of STC1 to chondrocytes and muscle cells suggests a role for this protein in chondrogenic and myogenic differentiation. PMID- 10810310 TI - Comparison of the effects of add-back therapy with various natural oestrogens on bone metabolism in rats administered a long-acting gonadotrophin-releasing hormone agonist. AB - The hypoestrogenic state induced by gonadotrophin-releasing hormone agonist (GnRHa) has been shown to be effective in the treatment of oestrogen-dependent disorders but to induce bone loss. Adding back low doses of oestrogen in GnRHa therapy has been proposed to prevent bone loss. The purpose of this study is to assess the efficacy of add-back therapy with different natural oestrogens such as oestrone (OE(1)), oestradiol (OE(2)) and oestriol (OE(3)). Three-month-old female rats (250 g) were subcutaneously administered microcapsules of leuprorelin acetate in doses of 1 mg/kg of body weight every 4 weeks. GnRHa therapy lasted 16 weeks, and pellets of OE(1), OE(2) or OE(3) (0.5 mg/pellet, 60 day release), as an add-back agent, were implanted at 8 weeks of treatment. At the end of treatment, GnRHa alone decreased bone mineral density of the femur and lumbar vertebrae, and increased serum levels of bone metabolic markers such as alkaline phosphatase and osteocalcin levels. As for cancellous bone histomorphometry, GnRHa decreased bone volume while it increased osteoid volume, osteoid surface, eroded surface, mineral apposition rate and bone formation rate. All the oestrogens tested prevented these changes caused by GnRHa therapy. GnRHa induced a significant increase in body weight and a marked reduction in uterine weight, which was not observed in OE(1) or OE(2) add-back group. Body weight and uterine weight of the OE(3) add-back group were the same as those of the GnRHa group. These findings indicate that GnRHa induces high turnover bone loss which can be prevented by concomitant administration of natural oestrogens such as OE(1), OE(2) and OE(3) to the same extent. In addition, OE(3) is unique in that it is much less effective than OE(1) and OE(2) in blocking body weight gain and in promoting growth of uterine tissues. Because of its tissue-selective actions, OE(3) could be considered as one of the most appropriate oestrogens used for GnRHa add-back therapy. PMID- 10810311 TI - Angiogenesis in pituitary adenomas - relationship to endocrine function, treatment and outcome. AB - Angiogenesis has been shown to be related to tumour behaviour, prognosis and response to treatment in many different tumour types. The aim of this study was to examine the relationship between angiogenesis and tumour behaviour and response to treatment in pituitary adenomas. The microvessel density (MVD) of pituitary tumours was assessed by counting blood vessels labelled with 3 different endothelial markers using antibodies to CD31, factor eight-related antigen and biotinylated Ulex europaeus (agglutinin I UEAI). One hundred and forty-two surgically removed pituitary adenomas (46 GH secreting, 6 microprolactinomas, 19 macroprolactinomas, 18 ACTH secreting and 53 functionless tumours) were carefully characterized and assessed. There was a significant negative correlation between age and MVD of GH secreting tumours (R(2)=33.8, P=0.005). Age was not related to MVD in other tumour types. Pre-treatment hormone production by the adenomas was related to MVD in prolactinomas (P<0.05), but not in GH secreting tumours. Invasive prolactinomas were significantly more vascular than non-invasive tumours (P<0.05). Drug treatment with metyrapone or bromocriptine did not appear to influence tumour angiogenesis. Surgical cure was more likely in macroprolactinomas and in ACTH secreting tumours with lower MVD. These results show that factors related to angiogenesis are very important in determining a number of clinical features of pituitary tumours, in particular the invasiveness of macroprolactinomas, the effect of age in tumours secreting GH and the outcome of surgical treatment in macroprolactinomas and ACTH secreting tumours. PMID- 10810312 TI - Vascular endothelial growth factor in the rat pituitary: differential distribution and regulation by estrogen. AB - Vascular endothelial growth factor (VEGF), an endothelial cell mitogen and permeability factor, participates in tumor angiogenesis, but less is known about its regulation or function in normal vascular homeostasis. In the uterus, which undergoes cyclic changes in its vasculature, VEGF is induced by estrogen. Since the pituitary gland contains highly permeable capillaries and is estrogen responsive, our objectives were to localize VEGF expression within the pituitary and to determine whether it is regulated by estrogen in both the pituitary and the somatolactotrope cell line, GH(3). Ovariectomized rats were injected with estradiol, and pituitaries and uteri were subjected to in situ hybridization or quantitative reverse transcription-polymerase chain reaction (RT-PCR). VEGF expression was strong and punctate in the neural lobe, weaker and diffuse in the anterior lobe and undetectable in the intermediate lobe. Two VEGF isoforms, 164 and 120, were detected in all tissues. In the posterior pituitary, VEGF expression was 3- to 6-fold higher than in the anterior pituitary or uterus and was unaltered by estrogen. In contrast, anterior pituitary VEGF was induced by estrogen within 1 h, peaked at 3 h, and returned to basal levels by 24 h. Similar dynamics, albeit 10-fold higher, were seen in the uterus. Translated VEGF proteins were detected by Western blot in both the anterior pituitary and uterus. GH(3) cells also showed a dose- and time-dependent induction of VEGF expression by estrogen. IN CONCLUSION: (1) VEGF expression is higher in the neural lobe than in the anterior lobe and is undetectable in the intermediate lobe, (2) the expression of VEGF164 and VEGF120 is rapidly upregulated by estrogen in the anterior pituitary but is unchanged in the posterior pituitary, and (3) the pituitary lactotrope cell line, GH(3), also increases VEGF expression in response to estradiol. PMID- 10810313 TI - Transforming growth factor-alpha stimulates proliferation of mammotrophs and corticotrophs in the mouse pituitary. AB - Oestrogen stimulates the proliferation of pituitary cells. The present study was designed to clarify the involvement of transforming growth factor-alpha (TGF alpha) in the oestrogen-induced growth of mouse pituitary cells in vitro. Anterior pituitary cells obtained from ICR male mice were cultured in a primary serum-free culture system. Proliferation of pituitary cells was detected by monitoring the cellular uptake of bromodeoxyuridine. Secretory cell types were immunocytochemically determined. Treatment with TGF-alpha (0.1 and 1 ng/ml) for 5 days stimulated cell proliferation. Since TGF-alpha binds to the epidermal growth factor (EGF) receptor, this action may be exerted through the EGF receptor. Oestradiol-17beta (OE(2), 10(-)(9) M) stimulated mammotrophic and corticotrophic cell proliferation. RG-13022, an EGF receptor inhibitor, inhibited the cell proliferation induced by EGF or OE(2), showing that the EGF receptor was involved in the growth response in mammotrophs and corticotrophs. Treatment with antisense TGF-alpha oligodeoxynucleotide (ODN) inhibited the cell proliferation induced by OE(2), but treatment with antisense EGF ODN did not. RT-PCR analysis revealed that OE(2) stimulated TGF-alpha mRNA and EGF receptor mRNA expression. These results indicate that TGF-alpha mediates the stimulatory effect of oestrogen on the pituitary cell proliferation in a paracrine or autocrine manner, and that EGF receptor expression is stimulated by oestrogen. PMID- 10810314 TI - Expression of activin A and its receptors in human pheochromocytomas. AB - Activin A (a homodimer of two activin betaA subunits) has been shown to induce the neuronal differentiation of rat pheochromocytoma PC12 cells. We studied activin A and its receptor gene expression in human pheochromocytomas in vivo and in vitro to clarify the potential involvement of activin A in the pathophysiology of these tumors. We first screened 20 pheochromocytomas and nine normal adrenal tissues for activin betaA mRNA expression. Northern blots hybridized with specific oligonucleotide probes detected weak signals for activin betaA transcripts in pheochromocytomas. Both type I and type II activin receptor (ActR I, ActR-IB and ActR-II) mRNA expression was also detectable in the pheochromocytoma tissues. In primary cultures of pheochromocytoma cells, expression of activin betaA mRNA was readily detectable by Northern blotting, and secretion of activin A into the conditioned medium was confirmed by an enzyme linked immunosorbent assay. The expression of activin betaA mRNA and secretion of activin A were induced by (Bu)(2)cAMP after 1 and 3 days of treatment (all P<0.05). A protein kinase inhibitor, staurosporine, inhibited the basal and (Bu)(2)cAMP-induced accumulation of activin betaA mRNA (P<0.05). In addition, induction of chromaffin phenotype by dexamethasone also inhibited the basal and (Bu)(2)cAMP-induced expression of activin A at both mRNA and protein levels (all P<0.05). In contrast, the expression of ActR-I and ActR-IB mRNAs was not affected by these agents in cultured pheochromocytoma cells. In summary, activin betaA subunit and activin receptors are expressed in human pheochromocytomas. Production of activin A in cultured pheochromocytoma cells is induced through the protein kinase A pathway, but reduced during chromaffin differentiation. Therefore, activin A may function as a local neurotrophic factor via an auto/paracrine manner in human pheochromocytomas. PMID- 10810315 TI - Glibenclamide but not other sulphonylureas stimulates release of neuropeptide Y from perifused rat islets and hamster insulinoma cells. AB - We have studied the effects of first and second generation sulphonylureas on the release of insulin and neuropeptide tyrosine (NPY) from hamster insulinoma tumour (HIT T15) cells and isolated rat islets. In the presence of 5.5 mmol/l glucose all sulphonylureas stimulated insulin release from the HIT cells (P<0.01 ANOVA, n> or =4) but only glibenclamide (GLIB, 10 micromol/l) stimulated the release of NPY (mean+/-s.e.m. control 11.1+/-1.3 vs GLIB 28.4+/-4.1 fmol/h per 10(6) cells, P<0001, n=16). In isolated perifused rat islets both glibenclamide (10 micromol/l) (control 3.5+/-0.3 vs GLIB 6. 3+/-0.2 fmol/min per islet, P<0.01, n=6) and tolbutamide (50 micromol/l) (control 4.7+/-0.1 vs TOLB 6.7+/-0.3 fmol/min per islet, P<0.01, n=6) enhanced glucose (8 mmol/l)-stimulated insulin release. However, only glibenclamide stimulated the release of NPY from the islets (control 3.4+/-0.8 vs GLIB 24.5+/-5 attomol/min per islet, P<0.01, n=6). Similar results were obtained in islets isolated from dexamethasonetreated rats. Glibenclamide treatment of HIT cells showed a prompt insulin release (10 min) while NPY secretion was slower (60 min), suggesting that internalization of the sulphonylurea is required to stimulate NPY release. Glibenclamide, the most common oral therapeutic agent in type 2 diabetes mellitus, is associated with release of the autocrine insulin secretion inhibitor, NPY. PMID- 10810316 TI - Plasma leptin determination in ruminants: effect of nutritional status and body fatness on plasma leptin concentration assessed by a specific RIA in sheep. AB - A specific leptin RIA was developed to assess concentrations of leptin in ovine plasma, and was shown to be efficient with bovine and caprine plasma. A specific, high-affinity antibody was generated against recombinant ovine leptin which, when used in a competitive leptin RIA, provided valid estimates of linearity (r=+0.989 0.998), recovery (102%), repeatability (13%) and limit of sensitivity (0.83 ng/ml for 100 microl sample size). Serial dilutions of five ovine, bovine or caprine plasma samples showed good linearity and parallelism with the recombinant ovine leptin standard curve. A comparison of this RIA was made with a commercial 'multi species' RIA kit using 56 ovine plasma samples. Major differences were found in assay sensitivity. Non-lactating, non-pregnant, ovariectomized ewes were fed a ration for 65 days which provided 90+/-9% (control; n=12) or 39+/-2% of maintenance energy requirements (underfed; n=16) in order to analyse the respective effects of body fatness (estimated by either an in vivo dilution technique or body condition scoring) and of nutritional status on plasma leptin concentration. There was a significant positive correlation between body fatness or body condition score and plasma leptin levels (r=+0.68, P<0.001 or r=+0.72, P<0.001 respectively). When concentrations of leptin were assessed over time, underfed ewes exhibited a dramatic reduction in plasma leptin values (-56%, P<0.001). These data provide strong evidence that, in sheep, the variations in plasma concentrations of leptin are related to variations in body fatness (35%) and, to a lesser extent, in nutritional status (17%). PMID- 10810317 TI - Naturally occurring scrapie is associated with a lower CBG binding capacity in ewes. AB - Naturally scrapie-affected ewes present a syndrome of hypercortisolism as evaluated by measuring total plasma cortisol concentrations. The objective of this study was to investigate the plasma protein binding of cortisol and to evaluate the concentration of the biologically active free fraction of cortisol in scrapie-affected ewes. Corticosteroid binding globulin (CBG) binding parameters were evaluated by equilibrium dialysis in 13 naturally scrapie affected ewes and nine healthy ewes, during two periods of the clinical evolution of the disease. The hypercortisolism of the scrapie-affected ewes was confirmed by a significant increase of the plasma 20 beta-dihydrocortisol and cortisone concentrations, while total cortisol concentrations, obtained from an isolated sample, did not differ between scrapie-affected and control ewes. The scrapie diagnosis was confirmed by histopathology. The CBG maximal capacity (B(max)) was two times lower in scrapie-affected ewes than in healthy ewes (37+/-32 nM and 73+/-28 nM respectively). The dissociation constant K(d) (8.8+/-3.7 nM and 9.8+/ 3.0 nM respectively) and the non-specific constant value of binding to albumin (1.13+/-0.18 and 1.14+/-0.23 respectively) did not differ significantly between diseased and control ewes. The significant increased concentrations of CBG-free cortisol (i.e. both albumin-bound and free cortisol fractions) in scrapie affected ewes indicates that total plasma cortisol concentration is not an appropriate index of pituitary-adrenocortical hyperactivity. In conclusion, ewes with naturally occurring scrapie display a syndrome of hypercortisolism associated with a lower CBG binding capacity which leads to an overexposure of glucocorticoid-sensitive targets to CBG-free cortisol. The physiopathological consequences of this overexposure on the development of the neurodegenerative process in prion disease are discussed. PMID- 10810318 TI - Aldosterone mediates angiotensin II-stimulated rat vascular smooth muscle cell proliferation. AB - Aldosterone, possibly locally generated, has been suggested to have a role in potentiating angiotensin II (AII)-stimulated hypertrophy of cultured vascular smooth muscle cells. To examine the possibility that aldosterone may mediate the proliferative actions of AII, rat aortic smooth muscle cells (RASMCs) in culture were treated with AII in the presence and absence of the specific AII type 1 receptor (AT1) antagonist, losartan, and aldosterone was assayed in culture medium extracts by radioimmunoassay. AII significantly enhanced aldosterone formation (at 10(-8) M: 123.8+/-14.85 vs control 71. 28+/- 8.71 fmol/10(5) cells, P<0.05; at 10(-7) M: 172.38+/-33.44, P<0.05), but not in the presence of losartan (at 10(-8) M: 53. 71+/-18.73, P>0.05; at 10(-7) M: 89.68+/-25.05, P>0.05). In other studies, the reverse transcriptase-polymerase chain reaction, performed on RNA extracted from RASMCs using aldosterone synthase (CYP11B2) specific primers, gave a single band of about 268 bp, consistent with that expected for the enzyme. Finally, using [(3)H]methylthymidine uptake as an index of cellular proliferation, tritium incorporation was increased in the AII-treated group at concentrations greater than 10(-10) M. The aldosterone antagonist, spironolactone (10(-5) M), inhibited the incorporation of [(3)H]thymidine into RASMCs stimulated by AII. These results suggest that locally generated aldosterone may mediate the effects of AII, acting via the AT1 receptor, in stimulating RASMC proliferation. PMID- 10810320 TI - Total IgE detection in paired cerebrospinal fluid and serum samples from patients with neurocysticercosis. AB - Neurocysticercosis (NC), the presence of Taenia solium metacestodes in tissues, is the most frequent and severe parasitic infection of the central nervous system. We investigated the presence of total IgE by an automated chemiluminescence assay in 53 paired cerebrospinal fluid (CSF) and serum samples from patients with NC (P) and in 40 CSF samples from individuals with other neurological disorders as the control group (C). Total IgE concentration ranged from 1.2 to 6.6 IU/ml (mean = 1.4 IU/ml, standard deviation-sd = 1.1 IU/ml) in 28.3% of CSF samples from the P group, a value significantly higher than for the C group ( pound1.0 IU/ml). The serum samples from the P group showed concentrations ranging from 1. 0 to 2330.0 IU/ml (mean = 224.1 IU/ml, sd = 452.1 IU/ml), which were higher than the normal value cited by the manufacturer (<100.0 IU/ml) in 32.1% of the samples. A significant difference was observed in CSF samples from the P and C groups (p = 0.005) and in serum samples from the P group compared to the normal value (p = 0. 005), with sera showing more frequent abnormal results. PMID- 10810319 TI - Fibrotic sequelae in pulmonary paracoccidioidomycosis: histopathological aspects in BALB/c mice infected with viable and non-viable paracoccidioides brasiliensis propagules. AB - Patients with paracoccidioidomycosis often present pulmonary fibrosis and exhibit important respiratory limitations. Based on an already established animal model, the contribution of viable and non-viable P. brasiliensis propagules to the development of fibrosis was investigated. BALB/c male mice, 4-6 weeks old were inoculated intranasally either with 4x10(6) viable conidia (Group I), or 6. 5x10(6) fragmented yeast cells (Group II). Control animals received PBS. Six mice per period were sacrificed at 24, 48, 72h (initial) and 1, 2, 4, 8, 12 and 16 weeks post-challenge (late). Paraffin embedded lungs were sectioned and stained with H&E, trichromic (Masson), reticulin and Grocot&tacute;s. During the initial period PMNs influx was important in both groups and acute inflammation involving 34% to 45% of the lungs was noticed. Later on, mononuclear cells predominated. In group I, the inflammation progressed and granulomas were formed and by the 12th week they fussed and became loose. Thick collagen I fibers were observed in 66.6% and 83.3% of the animals at 8 and 12 weeks, respectively. Collagen III, thick fibers became apparent in some animals at 4 weeks and by 12 weeks, 83% of them exhibited alterations in the organization and thickness of these elements. In group II mice, this pattern was different with stepwise decrease in the number of inflammatory foci and lack of granulomas. Although initially most animals in this group had minor alterations in thin collagen I fibers, they disappeared by the 4th week. Results indicate that tissue response to fragmented yeast cells was transitory while viable conidia evoked a progressive inflammatory reaction leading to granuloma formation and to excess production and/or disarrangement of collagens I and III; the latter led to fibrosis. PMID- 10810321 TI - ELISA test for the diagnosis of cysticercosis in pigs using antigens of Taenia solium and Taenia crassiceps cysticerci. AB - In the present study ELISA was standardized for the diagnosis of swine cysticercosis based on necropsy parameters and confirmed positive and negative control sera. Serum samples from pigs with other infections were also assayed to determine possible cross-reactions. Four antigens were assayed: from Taenia crassiceps vesicular fluid (VF-Tcra) and crude larvae extract (T-Tcra), and from Taenia solium extracts of scolex (S-Ts) and of larvae (T-Ts). A checkerboard evaluation of antigen, serum and conjugate dilutions, as well as the use of Tween 20 and skim cow milk in wash and blocking solution had a marked effect on improving ELISA performance. All the antigens showed a good performance, but VF Tcra was the best, with 96.0% and 80.0% sensitivities for cut-offs respectively at 2sd and 3sd, and corresponding specificities of 97.5% and 100.0%. Cross reactivity was observed only with hydatidosis and ascaridiosis. In view of the high performance observed, the ELISA test should be recommended for the diagnosis of cysticercosis in suspected swine in slaughterhouses and for the screening of cysticercosis in swine production. These results will support integrated measures of cysticercosis control throughout the chain of swine production, effectively contributing to public health. PMID- 10810322 TI - Prevalence of hepatitis B and C in the sera of patients with HIV infection in Sao Paulo, Brazil. AB - The objective of this study was to evaluate the prevalence of hepatitis B and C viruses in a group of HIV infected patients, followed at a single institution since 1996. 1,693 HIV positive patients (1,162 male, 531 female) were tested for HBV infection. Virological markers for HBV included HBsAg and total anti-HBc by ELISA. 1,457 patients (1,009 male, 448 female) were tested for HCV infection. Detection of HCV antibodies was carried out by ELISA. A sample of HCV antibody positive patients was tested for HCV by PCR to confirm infection. Of 1,693 patients tested for HBV, 654 (38.6%) and 96 (5.7%) were anti-HBc and HBsAg positive, respectively. Of 1, 457 patients tested for HCV, 258 (17.7%) were anti HCV positive. 82 of these patients were also tested by PCR and 81 were positive (98%). Of 1,411 patients tested for HBV and HCV 26 (1.8%) were positive for both viruses. PMID- 10810323 TI - Malaria vectors in the Brazilian amazon: Anopheles of the subgenus Nyssorhynchus. AB - Various species of Anopheles (Nyssorhynchus) were studied in the Amazon with the objective of determining their importance as malaria vectors. Of the 33 known Anopheles species occurring in the Amazon, only 9 were found to be infected with Plasmodium. The different species of this subgenus varied both in diversity and density in the collection areas. The populations showed a tendency towards lower density and diversity in virgin forest than in areas modified by human intervention. The principal vector, An. darlingi, is anthropophilic with a continuous activity cycle lasting the entire night but peaking at sunset and sunrise. These species (Nyssorhynchus) are peridomiciliary, entering houses to feed on blood and immediately leaving to settle on nearby vegetation. Anopheles nuneztovari proved to be zoophilic, crepuscular and peridomiciliary. These habits may change depending on a series of external factors, especially those related to human activity. There is a possibility that sibling species exist in the study area and they are being studied with reference to An. darlingi, An. albitarsis and An. nuneztovari. The present results do not suggest the existence of subpopulations of An. darlingi in the Brazilian Amazon. PMID- 10810324 TI - Antibody response in cattle after vaccination with inactivated and attenuated rabies vaccines. AB - Despite the absence of current official reports showing the number of cattle infected by rabies, it is estimated that nearly 30,000 bovines are lost each year in Brazil. In order to minimize the important economic losses, control of the disease is achieved by eliminating bat colonies and by herd vaccination. In this study, we compare the antibody response in cattle elicited by vaccination with an attenuated ERA vaccine (AEvac) and an inactivated-adjuvanted PV (IPVvac) vaccine. The antibody titers were appraised by cell-culture neutralization test and ELISA, and the percentage of seropositivity was ascertained for a period of 180 days. IPVvac elicited complete seropositivity rates from day 30 to day 150, and even on day 180, 87% of the sera showed virus-neutralizing antibody titers (VNA) higher than 0.5IU/ml. There were no significant differences between the VNA titers and seropositivity rates obtained with IPVvac in the two methods tested. AEvac, however, elicited significantly lower titers than those observed in the group receiving inactivated vaccine. In addition, the profiles of antirabies IgG antibodies, evaluated by ELISA, and VNA, appraised by cell-culture neutralization test, were slightly different, when both vaccines were compared. PMID- 10810325 TI - Evolutive behavior towards cardiomyopathy of treated (nifurtimox or benznidazole) and untreated chronic chagasic patients. AB - The aim of this work was to compare the evolution of chronic chagasic untreated patients (UTPs) with that of benznidazole or nifurtimox-treated patients (TPs). A longitudinal study from a low endemic area (Santa Fe city, Argentina) was performed during an average period of 14 years. Serological and parasitological analyses with clinical exams, ECG and X-chest ray were carried out. At the onset, 19/198 infected patients showed chagasic cardiomyopathy (CrChM) while 179 were asymptomatic. In this latter group the frequency of CrChM during the follow-up was lower in TPs compared with UTPs (3.2% vs 7%). Within the CrChM group, 2/5 TPs showed aggravated myopathy whereas this happened in 9/14 UTPs. Comparing the clinical evolution of all patients, 5.9% of TPs and 13% of UTPs had unfavourable evolution, but the difference is not statistically relevant. Serological titers were assessed by IIF. Titers equal to or lower than 1/64 were obtained in 86% of the TPs, but only in 38% of UTPs. The differences were statistically significant (geometric mean: 49.36 vs. 98.2). Antiparasitic assessment of the drugs (xenodiagnosis) proved to be effective. The low sensitivity in chronic chagasic patients must be born in mind. Despite treated patients showed a better clinical evolution and lower antibody levels than untreated ones, it is necessary to carry on doing research in order to improve therapeutic guidelines, according to the risk/benefit equation and based on scientific and ethical principles. PMID- 10810326 TI - Chromobacterium violaceum infection in Brazil. A case report. AB - We report the second case of infection with Chromobacterium violaceum that occurred in Brazil. A farm worker living in the State of Sao Paulo presented fever and severe abdominal pain for four days. At hospitalization the patient was in a toxemic state and had a distended and painful abdomen. Chest X-ray and abdominal ultrasound revealed bilateral pneumonia and hypoechoic areas in the liver. The patient developed failure of multiple organs and died a few hours later. Blood culture led to isolation of C. violaceum resistant to ampicillin and cephalosporins and sensitive to chloramphenicol, tetracyclin, aminoglicosydes, and ciprofloxacin. Autopsy revealed pulmonary microabscesses and multiple abscesses in the liver. The major features of this case are generally observed in infections by C. violaceum: rapid clinical course, multiple visceral abscesses, and high mortality. Because of the antimicrobial resistance profile of this Gram negative bacillus, for appropriate empirical antibiotic therapy it is important to consider chromobacteriosis in the differential diagnosis of severe community infections in Brazil. PMID- 10810327 TI - Cyclospora cayetanensis in sputum and stool samples. AB - We report the observation of acid-fast Cyclospora cayetanensis oocysts in a sputum sample. The patient, a 60 year-old, HIV negative man, was successfully treated for pulmonary tuberculosis during 1997. On February 1998, he was admitted to our center due to loss of weight, cough with purulent expectoration, dysphonia and a radiological picture of pulmonary fibrosis. Bacilloscopic study of sputum (negative for acid-fast bacilli) stained with Ziehl-Neelsen technique showed large (8-10 microm) spherical, acid-fast Cyclospora cayetanensis oocysts. No other pathogens were isolated on cultures from this sample or from laryngeal biopsy. Serial parasitologic studies showed C. cayetanensis and also eggs of Trichuris trichiura, Ascaris lumbricoides and Hymenolepis nana and of Entamoeba coli cysts. The patient lives in the outskirts of Buenos Aires in a brick-made house with potable water and works as builder of sewers. He travelled in several occasions to the rural area of province of Tucuman which has poor sanitary conditions. C. cayetanensis is an emergent agent of diarrhea and as far as we know this is the first time the parasite is observed in respiratory samples. PMID- 10810328 TI - Headache in an emergency room. PMID- 10810329 TI - Headache in an emergency room in Brazil. AB - CONTEXT: When experiencing a headache attack, Brazilian patients usually look for a primary care service, where they are seen by general clinicians. In the town of Ribeirao Preto, these clinicians routinely refer patients to the Emergency Room of the University Hospital. OBJECTIVE: The objective of this study was to evaluate the quality of primary care by analyzing retrospectively the medical records of patients with a complaint of headache seen in this emergency room during the year of 1996. DESIGN: retrospective study. SETTIING: Emergency Room of the Universital Hospital, Ribeirao Preto, Sao Paulo, reference unit. PARTICIPANTS: 1254 patients. The patients who sought the Emergency Room (ER) of the University Hospital of Ribeirao Preto, during the year of 1996 with a complaint of headache were studied retrospectively. MAIN MEASUREMENTS: Etiology, age, diagnosis, secondary cause, laboratory tests. RESULTS: Of the 1254 patients seen (61% women), 1190 (94.9%) were discharged after the administration of parenteral analgesics before they had spent 12 hours in the room. Only 64 (5.1%) patients remained for more than 12 hours. Of the patients who spent less than 12 hours in the room, 71.5% had migraine or tension type headache and did not require subsidiary exams for diagnosis. Of the patients who spent more than 12 hours in the room, 70.3% had secondary headaches. CONCLUSIONS: We conclude the primary care for headache is unsatisfactory in the Ribeirao Preto region. Many patients with primary headache are referred to tertiary care services, indicating the need for the dissemination of the diagnostic criteria of the International Headache Society to general practitioners. PMID- 10810330 TI - Comparison of vaginal wall sling and modified vaginal wall sling for stress urinary incontinence. AB - CONTEXT: There are several controversies about which is the best form of surgical treatment for stress urinary incontinence in women. The vaginal wall sling in its original and modified form were presented by Raz as new options for treatment of these conditions, but there is a lack of comparative clinical trials using both techniques. OBJECTIVE: To compare the effectiveness of the original and the modified vaginal wall sling. DESIGN: A comparative, prospective, non-randomized clinical trial. SETTING: Public and private health care units (Urology Division, Faculty of Medicine of the ABC Foundation, and Universidade Federal de Sao Paulo / Escola Paulista de Medicina). PARTICIPANTS: Twenty patients with anatomical and intrinsic sphincter deficiency stress urinary incontinence were surgically treated for evaluating the initial results of the vaginal wall sling, from February 5, 1994, to June 27, 1996. INTERVENTIONS: The patients were divided into two groups. Group A (n = 10) were treated with the original vaginal wall sling. Group B (n = 10) were treated with the modified vaginal wall sling. Both groups were statistically similar according to clinical and urodynamic parameters. MAIN MEASUREMENTS: Cure and complication rates. RESULTS: Follow-up ranged from 19 to 43 months (median = 28) for group A. The overall cure rate was 70%. Fifty per cent of the patients had urinary retention of 7 to 35 days. There were no major complications. Follow-up ranged from 14 to 26 months (median = 18) for Group B. The cure rate was 80%. Two patients had urinary retention of 7 and 55 days. There were no major complications. CONCLUSIONS: The vaginal wall sling is as effective as the modified vaginal wall sling but has a higher rate of urinary retention. PMID- 10810331 TI - Proliferating cell nuclear antigen expression in mucoepidermoid carcinoma of salivary glands. AB - CONTEXT: Among the cytological and morphological properties of mucoepidermoid carcinoma, one of the most important criteria for measuring its biological behavior and aggressiveness is cell proliferation. In this way, immunohistochemical markers of cell proliferation have been found to be useful in tumor classification and have formed part of the prognostic and therapeutic studies of these pathologies. OBJECTIVE: To analyze 11 cases of mucoepidermoid carcinoma (MEC) using the proliferation activity marker (PCNA) and to determine its relationship to the grade of malignancy of these tumors. DESIGN: Correlation study. SETTING: Head and Neck Surgery Service of Heliopolis Hospital, Sao Paulo, Brazil. SAMPLE: Slides of 11 cases of primary mucoepidermoid carcinomas of salivary glands were prepared according to routine techniques employed in the Oral Pathology Department of the Dentistry Faculty of Sao Paulo University, Brazil. They were fixed in a 10% formaldehyde solution and stained with hematoxylin and eosin. After this preparation the tumors were classified as low, intermediate and high grade of malignancy, according to the criteria established by Seifert & Sobin and Auclair, Goode & Ellis. The slides were sent for immunohistochemical processing to evaluate the positivity of proliferating cell nuclear antigen using the streptavidin biotin technique. MAIN MEASUREMENT: The correlation between proliferating cell nuclear antigen expression and the histological malignancy grade in mucoepidermoid carcinoma of salivary glands. RESULTS: there were 4 cases (36%) of low grade, 4 cases (36%) of intermediate grade and 3 cases (27%) of high grade of malignancy. After a comparative study between histological features and immunohistochemical analysis, significant differences were observed (P < 0.01) for low, intermediate and high grades: 16.04%, 26.98% and 56.98% of proliferating cell nuclear antigen expression in mucoepidermoid carcinoma, respectively. CONCLUSION: The proliferating cell nuclear antigen expression increases with the grade of malignancy in mucoepidermoid carcinoma of salivary glands. PMID- 10810332 TI - Post-mortem forensic identity testing: application of PCR to the identification of fire victim. AB - CONTEXT: DNA analysis has been used with success in the identification of carbonized corpses and victims of large accidents. The analysis requires relatives of crash victims to donate blood for analysis. The relatives are generally willing contribute to the identification by giving a blood sample. OBJECTIVE: To describe the use of the polymerase chain reaction (PCR) for genetic characterization of one victim extensively burned by fire. DESIGN: Case report. CASE REPORT: DNA was extracted from blood of the cardiac chamber, and 15 different loci (D1S80, ApoB, D17S30, D3S1744, D18S849, D12S1090, FGA, D7S820, D1S533, D9S304, HUMCSF1PO, HUMTPOX, HUMTHO1, amelogenin and HLA-DQA1) were analyzed using the PCR technique. Results from all loci typing of the corpse were then compared to that of his alleged biological parents, revealing a genetic compatibility. PMID- 10810333 TI - Lead poisoning after gunshot wound. AB - CONTEXT: Despite the absence of symptoms in the majority of patients carrying lead bullet fragments in their bodies, there needs to be an awareness of the possible signs and symptoms of lead intoxication when bullets are lodged in large joints like knees, hips and shoulders. Such patients merit closer follow-up, and even surgical procedure for removing the fragments. OBJECTIVE: To describe a patient who developed clinical lead intoxication several years after a gunshot wound. DESIGN: Case report. CASE REPORT: A single white 23-year-old male, regular job as a bricklayer, with a history of chronic alcohol abuse, showed up at the emergency department complaining of abdominal pain with colic, weakness, vomiting and diarrhea with black feces. All the symptoms had a duration of two to three weeks, and had been recurrent for the last two years, with calming during interval periods of two to three weeks. Abdominal radiograms showed a bullet lodged in the left hip, with a neat bursogram of the whole synovial capsule. A course of chelating treatment using calcium versenate (EDTACaNa2) intravenously was started. After the chelation therapy the patient had recurrence of his symptoms and a radical solution for the chronic mobilization of lead was considered. A hip arthroplasty procedure was performed, leading to complete substitution of the left hip. PMID- 10810334 TI - American Spinal Injury Association 26th annual meeting. Chicago, Illinois, USA. April 14-16, 2000. Abstracts. PMID- 10810335 TI - Association between the level of ERCC-1 expression and the repair of cisplatin induced DNA damage in human ovarian cancer cells. AB - Nucleotide excision repair (NER) is responsible for the repair of platinum-DNA lesions. ERCC-1 is a critical gene within the NER pathway, and cells without a functional ERCC-1 do not repair cisplatin-caused DNA damage. The present study was therefore designed to evaluate the relationship between the expression of ERCC-1 and the repair of cisplatin-induced DNA adducts in human ovarian cancer cells in vitro. One hour exposure of MCAS cells to cisplatin yielded an approximately two-fold increment in the levels of ERCC-1 mRNA and ERCC-1 protein, as determined, respectively, by Northern and Western blottings. In addition, nuclear run-on assay showed that ERCC-1 gene transcription rate was increased to about the same extent as steady-state ERCC-1 mRNA and protein, in response to cisplatin treatment. However, the levels of ERCC-1 mRNA, ERCC-1 protein, and ERCC 1 transcript in MCAS cells are two-fold lower than those in A2780/CP70 cells, as previously reported. Furthermore, the repair of cisplatin-DNA adducts in MCAS cells, as measured by atomic absorption spectrometry, is also nearly two-fold less than that in A2780/CP70 cells, indicating a strong association between the level of ERCC-1 expression and the activity of excision repair in these two human ovarian tumor cell lines. These results suggest that ERCC-1 may be a useful marker to monitor the repair of platinum-DNA damage in tumor cells, and further highlight that potential pharmacological approaches which specifically inhibit ERCC-1 expression may increase cellular sensitivity to cisplatin. PMID- 10810336 TI - Inhibition of MDA-MB-231 human breast tumor xenografts and HER2 expression by anti-tumor agents GAP31 and MAP30. AB - GAP31 (Gelonium protein of 31 kDa) and MAP30 (Momordica protein of 30 kDa) are agents isolated from the medicinal plants Gelonium multiflorum and Momordica charantia, respectively. The current study was conducted to investigate the efficacy of GAP31 and MAP30 on estrogen-independent and highly metastatic human breast tumor MDA-MB-231 both in vitro and in vivo. The effect of these agents on the expression of breast tumor antigen HER2 (also known as neu or as c-erbB 2) was also examined. Treatment of MDA-MB-231 breast cancer cells with GAP31 and MAP30 resulted in inhibition of cancer cell proliferation as well as inhibition of the expression of HER2 gene in vitro. When MDA-MB-231 human breast cancer cells were transferred into SCID mice, the mice developed extensive metastases and all mice succumbed to tumor by day 46. Treatment of the human breast cancer bearing SCID mice with GAP31 or MAP30 at 10 micrograms/injection EOD for 10 injections resulted in significant increases in survival, with 20-25% of the mice remaining tumor free for 96 days. Thus, anti-tumor agents GAP31 and MAP30 are effective against human breast cancer MDA-MB-231 in vitro and in vivo. These agents may therefore be a potential therapeutic use against breast carcinomas. PMID- 10810337 TI - Ectopic expression of truncated Sp1 transcription factor prolongs the S phase and reduces the growth rate. AB - The role of the transcription factor Sp1 in cell growth and survival was investigated by induced expression of its DNA-binding C-terminal fragment. Transfection of a constitutively expressed Sp1-170C plasmid construct into HeLa cells failed to produce viable clones, suggesting that this peptide interferes with cell growth. However, transfection with the muristerone A-inducible vector system produced four clones with low levels of expression of Sp1-170C. Muristerone A transiently induced higher levels of Sp1-170C, and this was accompanied by a reduced growth rate and prolongation of the S phase of the cell cycle. This is the first report that a dominant negative Sp1 can affect the cell cycle. PMID- 10810338 TI - Modulation of MCF-7 breast cancer cell signal transduction by linoleic acid and conjugated linoleic acid in culture. AB - The effects of modifying membrane fatty acid composition on cell growth, phospholipase C (PLC) and protein kinase C (PKC) activities, and prostaglandin E2 (PGE2) secretion were investigated. Hormone responsive MCF-7 human breast cancer cells were incubated in a serum-free medium containing epidermal growth factor and supplemented with physiologic concentrations (0.18-1.78 x 10(-5) M) of linoleic acid (LA) or conjugated linoleic acid (CLA). Linoleic acid stimulated cancer cell growth, while CLA was inhibitory. Supplementation with LA or CLA altered cell membrane composition. Linoleic acid stimulated PLC activity with or without GTP gamma (S), and tended to increase membrane PKC activity. However, CLA supplementation did not modify membrane PLC or PKC activity. Prostaglandin E2 secretion was not influenced by LA or CLA. These data show that growth inhibition by CLA was not mediated through PLC-, PKC- or PGE2-dependent signal transduction pathways, suggesting that another inhibitory mechanism may be involved. Although biological differences appeared to be modest (5-20% of control), the fact that LA and CLA treatment resulted in significant biological effects at physiologic concentrations is relevant, since most human cancers require years to develop. PMID- 10810339 TI - Sestamibi accumulation in human nasopharyngeal carcinoma cell lines in vitro. AB - Sestamibi imaging is used to assess the staging of nasopharyngeal carcinoma (NPC), and to monitor response to therapy. Sestamibi was added to single-cell suspensions of the NPC cell lines CNE-1 and CNE-2Z, and aliquots were removed over 60 min and centrifuged to determine cell-associated radioactivity. Sestamibi accumulation reached similar plateau values in both cell lines within 30 min of addition. Saturating concentrations of P-glycoprotein (Pgp) modulators increased accumulation by 1.4-1.7 fold over controls. Hyperpolarization of the mitochondrial membrane with nigericin increased accumulation by 2.6-3.4 fold. In contrast, depolarization of the plasma membrane with isotonic high potassium buffer reduced accumulation to 70-75% of control values, and additional depolarization of the mitochondrial membrane with valinomycin further reduced accumulation to 21-29% of control levels. These results indicate that both cell lines contain a modest level of Pgp activity and that both are capable of further polarization of the mitochondria. This suggests that mitochondrial hyperpolarization is not the complete explanation for high accumulation of sestamibi in NPC. PMID- 10810340 TI - EORTC receptor and biomarker study group report analytical and technical evaluation of a thymidine kinase radio-enzymatic assay in breast cancer cytosols. AB - BACKGROUND: High thymidine kinase (TK) activity in cancer cells could counteract adjuvant chemotherapy directed at the inhibition of de novo DNA synthesis. TK is an independent prognostic factor in breast cancer patients receiving adjuvant chemotherapy. MATERIAL AND METHODS: In this paper, we describe the effects of extraction and dilution buffer composition on TK enzymatic activity values obtained in breast cancer cytosols with the Prolifigen serum TK-REA kit (Sangtec Medical, Sweden). RESULTS: The addition of MgCl2 and ATP early in the assay, preferably during the extraction of tumor tissue, seems critical to stabilise the enzyme. Furthermore, the use of normal calf serum to dilute both standards and samples is necessary to obtain satisfactory parallelism between TK values in serial dilutions of breast cancer cytosols. CONCLUSION: Based on the data reported here, the manufacturer has changed the cytosol diluent composition and is adding a specific cytosol assay insert to the Prolifigen TK-REA kit. As evidenced by the laboratory reproducibility, these modifications to the serum assay led to an adequate, standardized protocol for analyzing TK activity in breast tumor cytosols. PMID- 10810341 TI - The effects of glycosylation inhibitors on the proliferation of a Wilms tumour cell line. AB - We have examined the effects of different glycosylation inhibitors on the proliferation of a human Wilms tumour derived cell line WCCS-1. It was found that two compounds that specifically inhibit distal steps in the glycosylation chain (swainsonone and castanospermine) only exerted marginal effects on cell multiplication and survival. In contrast, a proximal inhibitor (tunicamycin) efficiently increased necrosis in a dose dependent fashion. It is shown that this cell death was accompanied by a marked decrease in the incorporation of glucosamine, but rather unexpectedly, only caused a limited inhibition of de novo protein synthesis. Moreover, the entrance into S-phase was virtually unchanged in the cells surviving the exposure to tunicamycin. The effects of tunicamycin on cell multiplication and survival could not be reversed by concomitant addition of mevalonate as has been shown in other cell lines. Taken together this data suggests that tunicamycin does not operate in a cell cycle specific manner in Wilms tumour cells. PMID- 10810342 TI - Induction of senescence-like phenotype and loss of paclitaxel sensitivity after wild-type p53 gene transfection of p53-null human non-small cell lung cancer H358 cells. AB - The p53 gene plays an important role in the regulation of cell-cycle progression and apoptosis. Recent studies have implicated p53 in determining cell fate, and shown that p53 status is associated with cellular sensitivity to anticancer agents. However, the role of p53 in paclitaxel-induced cytotoxicity remains unclear. Here we show that the induction of exogenous wild-type (wt) p53 genes in p53-null human NSCLC H358 cells via transient gene transfection with cationic liposome-wt p53 complexes resulted in a typical senescence-like phenotype. In short, cell growth was reduced, homeostasis occurred, cell morphology became enlarged and flat, the cell cycle was arrested at G1 phase, cyclin B1 and cdc2 expression was down-regulated, and DNA synthesis was suppressed. The sensitivity of wt p53-transfected cells (H358/p53) to paclitaxel was approximately 3-fold lower than that of H358 cells. Paclitaxel treatment gradually and significantly blocked cell-cycle progression at G2/M phase and increased the accumulation of cyclin B1 and cdc2 in H358 cells. In contrast, the same treatment slightly arrested the cell cycle at G2/M phase and slightly elevated cyclin B1 expression in H358/p53 cells. The rate of uptake and efflux of paclitaxel was not significantly different between H358 and H358/p53 cells, indicating that the reduction in cellular sensitivity caused by p53 transfection was not due to alterasion in intracellular drug concentration. Together, our findings suggest that the induction of exogenous wt p53 gene expression in cells lacking p53 function can trigger the senescence program and that loss of sensitivity to paclitaxel by p53-transfected cells may be associated, at least in part, with the induction of a senescence-like phenotype. PMID- 10810343 TI - Development of an intracerebral glioma model in whole body irradiated hairless rats. AB - AIMS: To study the in vivo radiosensitivity of malignant gliomas, an animal glioma model was developed using the implantation of glioma cell lines into the brain of the Hairless rat (a mutant from the Sprague-Dawley strain, characterised by its complete absence of hair). METHODS: 10(6) malignant cells were suspended in 10 microliters phosphate buffered saline (PBS) and injected at a 4 microns depth into the left frontal lobe of an anaesthetised animal through a small craniotomy hole without opening the dura mater. The glioma cell line C6 (obtained from a chemically-induced rat glioblastoma) was introduced into 11 animals, and the human glioblastoma line G5 into 12 animals. RESULTS: The tumour take was checked using histological criteria. It was poor: 0% for the G5 line and only 27.3% for the C6 line. To improve the tumour growth rate, rats were subjected to a single dose (3.5 Gray) total body irradiation, 24 hours prior to injection, causing a marked immunosuppression. 84.6% of the rats grafted with the C6 line then produced tumours. Similar results (75% tumour take) were obtained using a stereotactic inoculation of the tumour cells. CONCLUSIONS: Thanks to the contribution of whole body irradiation, an animal intracerebral glioma model was establish, which can be used for clinical and biological studies. PMID- 10810344 TI - Correlation between iododeoxyuridine and MIB-1 labelling index in gastric carcinoma and adjacent normal gastric tissue. AB - BACKGROUND: In vivo labelling with synthetic thymidine analogues, such as Iododeoxyuridine (IdU) and Bromodeoxyuridine (BrdU), has frequently been used to estimate tumour proliferation. However, this method requires intravenous administration of IdU or BrdU, thymidine analogues that are potential mutagens. Recently, the monoclonal antibody MIB-1 has been developed, recognizing the Ki-67 nuclear antigen, which is associated with cell cycle proliferation and is found throughout the cell cycle (G1, S, G2 and M phases), but not in resting (G0) cells. We studied the correlation between the MIB-1 labelling index (LI) and the IdU labelling index in normal and malignant gastric tissue. PATIENTS AND METHODS: Twenty patients with gastric cancer received an intravenous injection of IdU (200 mg/m2) before surgery. Specimens were obtained from gastric carcinoma and adjacent normal gastric tissue. The samples were fixed in formalin and immunohistochemical analyses of IdU LI and MIB-1 LI were performed. The LI was defined as the percentage of labelled nuclei of 5000 nuclei counted. RESULTS: The IdU LI ranged from 3.3% to 18.2% in gastric carcinoma and from 0.5% to 5.6% in adjacent normal gastric mucosa, whereas the MIB-1 LI ranged from 4.2% to 46.0% in gastric cancer and from 1.3% to 25.1% in adjacent normal gastric mucosa. Comparison of IdU LI with MIB-1 LI, using the Spearman rank correlation coefficient test showed a significant correlation between IdU LI and MIB-1 LI in normal gastric tissue (r = 0.63, p < 0.05). However, in gastric carcinoma no significant correlation was found between either proliferation marker (r = 0.07, N.S.). CONCLUSION: MIB-1 accurately reflects the in vivo IdU LI in normal gastric tissue, whilst in gastric carcinoma the MIB-1 LI does not seem to be a substitute for the in vivo IdU LI. PMID- 10810345 TI - A nude mouse model of massive liver and lymph node metastasis of human colon cancer. AB - Liver and lymph nodes metastasis are the main causes of treatment failure for advanced colon cancer. However, currently-available animal models of human colon cancer do not demonstrate sufficient metastasis to represent highly malignant colon cancer that extensively metastasizes to these sites. A liver metastasis from a patient with highly malignant, poorly differentiated adenocarcinoma of the colon was established in nude mice by surgical orthotopic implantation to the mouse colon. The human origin of the tumor growing in nude mice was confirmed by in situ hybridization of human DNA. After 20 passages from the first implantation, massive liver and lymph nodes metastasis, occurred in 100% of the transplanted animals. Lymph nodes metastasis were found at the sites of lymph node drainage of the liver: celiac, portal and mediastinal lymph nodes. However no mesenteric and retroperitoneal nodes or lung tissue metastases were observed. Our data suggest that the mediastinal, celiac and hepatic lymph nodes metastases are derived form the liver metastasis, confirming the concept of metastasis of metastases or "remetastasis" of colon cancer. PMID- 10810346 TI - 1,25-dihydroxyvitamin D3 dissolved in lipiodol produces a sustained antiproliferative effect in the human hepatoblastoma cell line HepG2. AB - The steroid hormone 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] has potential to be used as an anti-tumor agent, but its clinical application has been restricted by the strong systemic calcemic activity. Regional administration of the drug dissolved in lipiodol, might be a way of selectively delivering high concentrations of the drug to lipiodol avid tumor cells without causing systemic side effects. In acute (1 day treatment) and chronic (5 days treatment) experiments, efficacy of the drug dissolved in ethanol (control) or lipiodol and subsequently diluted in the culture medium was tested in vitro against the hepatoblastoma cell line HepG2. Using [3H]thymidine incorporation and cell count, antiproliferative effects of 1,25-(OH)2D3 dissolved in the two different solvents was compared. Microscopic examination of cells exposed to the lipiodol containing media revealed intra-cellular presence of the oil in abundance. Chronic treatment of cells with either formulation of 1,25-(OH)2D3 resulted in profound inhibition of cell proliferation. However, exposure of cells to 1,25-(OH)2D3 in lipiodol was followed by significantly greater and lasting inhibition of cell proliferation in both acute and chronic studies. These results indicate that, 1,25-(OH)2D3 dissolved in lipiodol probably acts as a sustained release drug depot formulation, in which case it could have some potential for the regional treatment of liver tumors. PMID- 10810347 TI - Anomalies of the TGF-beta postreceptor signaling pathway in ovarian cancer cell lines. AB - Transforming growth factor-beta (TGF-beta) can cause cell cycle arrest, terminal differentiation, or apoptosis in most normal epithelial cells, whereas most malignant cell lines are resistant to TGF-beta. Mechanisms of resistance to TGF beta caused by modulation of cell cycle regulators and/or inactivation of components of the TGF-beta signaling transduction pathway such as C-myc and Smad4 have been demonstrated in human pancreatic cancer and squamous cell carcinoma cell lines. But, this has not been shown in ovarian cancer. To investigate the potential association between loss of sensitivity to TGF-beta and expression status of transforming growth factor receptor II (T beta RII), Smad4, CDC25A and C-myc in fourteen cell lines derived from ovarian cancer, the expression levels of these genes were examined by semi-quantitative RT-PCR. Normal ovarian surface tissues were used as controls. Expression of T beta RII was detectable in all of fourteen cell lines. Expression of Smad4 was decreased in ten cell lines and nine cell lines overexpressed CDC25A, compared to normal controls. CDC25A gene was overexpressed in 88% (8/9) of tumorigenic cell lines as determined by xenografts in nude mice, and only in 20% (1/5) of non-tumorigenic cell lines (P < 0.05). C myc was not overexpressed in any of these cell lines. The loss of sensitivity to TGF-beta of cell lines derived from ovarian cancers may be related to (1) a decreased expression of Smad4, which mediates TGF-beta induced growth inhibition; and/or (2) an overexpression of CDC25A. This overexpression correlates with increased tumorigenicity of ovarian cancer cell lines. The loss of sensitivity to TGF-beta is not associated with a lack of T beta RII. PMID- 10810348 TI - The study of gemcitabine in combination with other chemotherapeutic agents as an effective treatment for prostate cancer. AB - BACKGROUND: Gemcitabine has demonstrated clinical activity against several common cancers. Our studies examine the ability of gemcitabine, both alone and in combination with other chemotherapeutic agents, to inhibit the in vitro and in vivo growth of several prostate cancer cell lines. MATERIALS AND METHODS: Cultures of LNCaP, PC-3 or MLL cells were exposed to either gemcitabine or other appropriate agents for specified amounts of time. Cells were lysed and nuclei counted utilizing a Coulter Counter. For in vivo experiments, animals were injected with 1 x 10(5) MLL cells subcutaneously into the right flank. Animals were treated as indicated for 14 days. Tumors were then excised, weighed and measured. RESULTS: In both human (PC-3 and LNCaP) and rat prostate (MLL) cancer cell lines our studies demonstrated gemcitabine had a strong effect in vitro, with an IC50 of approximately 500 nM in the human lines and 10 nM in MLL cells. In vivo, studies using the Dunning prostate cancer model in Copenhagen rats resulted in a dose response inhibition of tumor growth, with an 80% decrease in tumor size in rats treated with gemcitabine at 10 mg/kg. CONCLUSIONS: Our results demonstrated the potent activity of gemcitabine against prostate cancer in the Dunning rat model and suggest the addition of paclitaxel may not aid in this activity. PMID- 10810349 TI - Immunocytochemical detection of hMSH2 and hMLH1 expression in oral melanoma. AB - BACKGROUND: The DNA mismatch repair system (MMR) plays an important role in the maintenance of genomic stability. To date few studies have been performed on hMSH2 and hMLH1 expression and melanoma of the head and neck region. A study of two cases revealed no mutations of the mismatch repair genes hMSH2 and hMLH1. MATERIALS AND METHODS: To verify the possibility of implication of hMSH2 and hMLH1 alterations in melanocytic cancerogenesis, the authors examined the protein expression pattern of hMSH2 and hMLH1 by immunohistochemistry in 9 paraffin embedded oral melanoma. RESULTS: One case (11%) showed nuclear positivity for hMSH2, 3 cases (33%) showed cytoplasmic positivity, and five cases (55%) showed no staining in the tumoral cells, even if normal squamous epithelium available in this section showed a nuclear positivity. Four cases (44%) showed no hMLH1 staining in the tumoral cells, even if normal squamous epithelium available in this section showed a nuclear positivity. Two cases (22%) showed nuclear positivity, and three cases (33%) showed cytoplasmic positivity. CONCLUSIONS: The analysis of mismatch repair genes can be a new molecular diagnostic tools for the detection of patients at high risk of developing melanoma and other neoplasia, or metastasis and recurrences. PMID- 10810350 TI - Human bladder carcinoma cells with an unusual pattern of in vitro growth: transition from nonproliferative spheroids to active monolayer growth upon interaction with tumor-derived fibroblasts. AB - A new transitional cell carcinoma cell line, BCCA-1, derived from a primary urinary bladder carcinoma, was characterized with respect to the growth patterns of in vitro culture, xenotransplantability in SCID mice and immunophenotypic profile. The most unusual finding was a strong tendency of forming many aggregates (multicell spheroids) in the first few days of flask cultures, followed by the attachment of spheroids to monolayer fibroblasts, which came along from stroma of the same tumor. Unlike those reported tumor spheroids whose peripheral layers contained proliferative cells, BCCA-1 spheroids rarely contained mitotic cells. The three-dimensional architecture of BCCA-1 spheroids drastically changed by the attachment of spheroids to fibroblasts, from which epithelial tumor cells spread; this was accompanied by pseudopodia formation and highly aggressive growth of tumor cells. As the fibroblasts degenerated due to overgrowth, tumor cells started to aggregate by retracting their pseudopods and forming many semi-attached spheroids, which eventually detached from the sheet of degenerated fibroblasts. BCCA-1 produced solid tumors as xenografts in SCID mice by subcutaneous injection with as low as 5 x 10(6) cells, suggesting malignant nature of these cells. Immunostaining revealed the expression of MHC-class I, S100 protein, cytokeratin CK7 and CK20, beta-HCG, CEA, epithelial membrane antigen, Le(y) and folate-binding protein by this tumor. While the biological significance of spheroid formation of this kind by BCCA-1 cells remains unclear, it may represent a protection mechanism, by which TCC cells could sustain their viability under unfavorable culture conditions, but proliferate when the conditions became improved, such as the presence of fibroblasts. Our results point to the importance of tumor-associated stromal fibroblasts in TCC tumor progression. Further mechanistic studies to elucidate the mechanism involved in the stromal cell contact mediated-activation of TCC cells in this model system are warranted. PMID- 10810351 TI - Papaverine combined with prostaglandin E2 synergistically induces neuron-like morphological changes and decrease of malignancy in human prostatic cancer LNCaP cells. AB - We are interested in the possibility of new prostate cancer therapy that would control tumor malignancy via the induction of terminal cell differentiation. Here, we investigated the combined effect of various cAMP reagents on LNCaP human prostate carcinoma cells. Papaverine and prostaglandin E2 (PGE2), combined synergistically induced morphological changes. Electron microscope study suggested that cells treated with both reagents become like neuroendocrine cells. We then investigated the effect of both reagents on proliferation and malignancy of LNCaP cells. The malignancy of cells was analyzed by soft agar colony-forming assay and an in vitro invasion assay. Proliferation and malignancy of LNCaP cells treated with both reagents were significantly decreased in comparison to the proliferation and malignancy of untreated cells. Furthermore, the expression of oncogenes such as c-myc and Bcl-2 was suppressed in differentiated LNCaP cells. These results suggest that papaverine combined with PGE2 can synergistically induce neuronal differentiation as well as decrease the malignancy of human prostatic cancer LNCaP cells. PMID- 10810352 TI - Influence of bovine calf serum on the intracellular accumulation and retention of adriamycin. AB - In this study, we observed the influence of bovine calf serum (BCS) on the intracellular accumulation of adriamycin (ADR) in Ehrlich ascites tumor cells (wild type EAT cells) and their ADR-resistant strain. The ADR accumulation treated in the medium not containing BCS was 3-fold of that containing 10% BCS. ADR incorporated into cells was not effluxed in the absence of the BCS, while it was rapidly effluxed for 30 minutes to about 20% of the amount incorporated into the cells in the presence of 10% BCS in both strains. This suggests that BCS remarkably decreases the ADR accumulation due to the promotion of ADR efflux. Further, the ADR accumulation decreased with increasing concentrations of BCS in both strains. This indicates that BCS promotes the ADR efflux, depending on its concentrations. BCS was heated at 100 degrees C for 7 minutes and its protein contents were degenerated. Its influence in the ADR accumulation of wild type and ADR-resistant strain was apparent. The ADR accumulation decreased with increasing concentrations of heated BCS in both strains, as well as in the untreated BCS. Thus, the protein content of the BCS did not affect ADR accumulation. Therefore, it is necessary to investigate which components of BCS affect the promotion of ADR efflux. PMID- 10810353 TI - Telomerase activity is down regulated via decreases in hTERT mRNA but not TEP1 mRNA or hTERC during the differentiation of leukemic cells. AB - BACKGROUND: Recent studies have determined several telomerase-associated molecules, but the precise mechanisms regulating telomerase activity by those molecules has not been fully understood. MATERIALS AND METHODS: The telomerase activity was determined by TRAP assay. Using TaqMan RT-PCR, the quantitative and kinetic values of mRNA expression of the three telomerase-associated molecules were examined in HL60 cells differentiated with tumor necrosis factor mutant and all-transretinoic acid. RESULTS: The levels of telomerase activity in leukemic cell lines, leukemic cells from patients, and normal peripheral blood cells were distributed over a very wide range. Human telomerase catalytic subunit (hTERT) mRNA expression declined to nearly undetectable levels more rapidly than the inhibition of telomerase activity after treatment with these reagents in HL60 cells. Telomerase-associated protein (TEP1) mRNA increased approximately 6-fold over its level in untreated cells. The levels of human telomerase RNA component (hTERC) also increased approximately 2.7-fold at 5 days after treatment. CONCLUSIONS: These results suggest that telomerase activity is down-regulated mainly via decreases in hTERT, but not TEP1 and hTERC expression during the differentiation of leukemic cells. PMID- 10810354 TI - Induction of apoptosis in human gastric cancer by sodium butyrate. AB - BACKGROUND: Novel therapeutic agents are needed in the adjuvant treatment of gastric cancer. The differentiating agent sodium butyrate (NaBT) inhibits the growth of colon cancer cells; its effects on gastric cancers are not known. The purpose of our study was to characterize the effects of NaBT on human gastric cancer. MATERIAL & METHODS: The human gastric cancer, SIIA, was treated with NaBT (5 mM) for 12-72 h. Cell number, viability and death were measured. Expression levels of the tumor-suppressor protein, p53, the cell-cycle inhibitors, p21Waf1/Cip1 and p27Kip1, and the pro-apoptotic proteins, Bax, Bak, and Bik, were determined. RESULTS: NaBT significantly inhibited SIIA gastric cancer cell proliferation in a time-dependent fashion by a process involving the induction of apoptosis. Treatment with NaBT was associated with increased expression levels of p21Waf1/Cip1 p27Kip1, Bax, Bak, and Bik. CONCLUSIONS: NaBT triggers growth arrest and apoptosis in the human gastric cancer SIIA potentially through the induction of the cell-cycle inhibitors, p21Waf1/Cip1 and p27Kip1, and the proapoptotic genes, Bax, Bak, and Bik. NaBT may be an effective adjuvant agent in the treatment of gastric cancer. PMID- 10810355 TI - Antiproliferative activity and mechanism of action of DZ-3358, a novel pyrimidinyl pyrazole derivative. AB - The novel pyrimidinyl pyrazole derivative, 1-[5-methyl-1-(2-pyrimidinyl) -4 pyrazolyl]-3-[4-(3-chlorophenyl)-1-piperazinyl]-1-trans-propene hydrochloride (DZ 3358), was found through random screening to exhibit anti-proliferative activity against human and murine cancer cell lines. DZ-3358 induced the mitotic arrest of P388 murine leukemia cells at 0.4 microgram/ml in a time-dependent manner, and inhibited porcine tubulin polymerization. Furthermore, using immunofluorescence techniques, we observed microtubule formation in NUGC-3 human gastric cancer cells treated with DZ-3358. Microtubule formation was disordered, similar to that which occurred when such cells were treated with colchicine. These findings suggest that the mechanism of the anti-proliferative effect of DZ-3358 is the inhibition of mitosis due to the blocking of tubulin polymerization. The differential pattern of growth inhibitory concentrations of DZ-3358 against a series of cancer cell lines was compared with that of colchicine and vincristine, and no correlation was found with the pattern of these two drugs. DZ-3358 may be useful as a new type of tubulin inhibitor and anti-cancer drug. PMID- 10810356 TI - Carcinoma-associated antigens MK-1 and CEA in urological cancers. AB - BACKGROUND: The MK-1 antigen, recognized by monoclonal antibody FU-MK-1, is widely associated with human carcinomas. However, the expression and distribution of MK-1 in urological cancers is not well known. MATERIALS AND METHODS: We examined the expression of MK-1 in 10 urological tumor cell lines using flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR) and in 15 cancer tissue specimens by immunohistochemical staining, and then compared it with that of carcinoembryonic antigen (CEA). RESULTS: When analyzed by flow cytometry, MK-1 was positive in 2 out of 3 bladder, 3 out of 3 prostate and one out of 4 renal tumor cell lines, whereas CEA was negative in all the 10 tumor cell lines. RT-PCR confirmed the presence of MK-1 mRNA in all the six MK-1 positive tumor cell lines. An immunohistochemical study demonstrated that MK-1 was positive in 2 out of 5 bladder, 2 out of 5 prostate and one out of 5 renal cancer tissues. Again, however, CEA was negative in all the 15 urological cancer tissues tested. CONCLUSION: These findings suggest that MK-1 seems to be a useful biological marker for malignant urological tumors, especially in cases of bladder and prostate cancer. PMID- 10810357 TI - Doxorubicin-gallium-transferrin conjugate overcomes multidrug resistance: evidence for drug accumulation in the nucleus of drug resistant MCF-7/ADR cells. AB - Development of multidrug resistance (MDR) in cancer cells decreases net doxorubicin (ADR) uptake as a result of increased efflux, increased intracellular sequestration, and decreased membrane permeability. In this study, we investigated whether conjugation of ADR to transferrin (Tf) could overcome MDR in breast cancer cells. The multidrug resistant MCF-7/ADR breast cancer cell line was over 1000-fold more resistant to ADR, than its parental MCF-7 cell line, as determined by 3[H]-thymidine assay. The MCF-7/ADR cell line also expressed both MDR1 and MRP genes, as detected by RT-PCR. The ADR was coupled using a glutaraldehyde technique to human transferrin saturated with either ferric chloride (Fe-Tf) or gallium nitrate (Ga-Tf). These conjugates were tested for cytotoxicity on both MCF-7 and MCF-7/ADR cells after 6 days of incubation. The doxorubicin-gallium-transferrin conjugate (ADR-Ga-Tf) exhibited approximately the same inhibitory effect as ADR on MCF-7 cells with IC50s of 2.34 x 10(-3) microM and 1.42 x 10(-3) microM, respectively. However in MCF-7/ADR cells ADR-Ga-Tf reversed resistance to free ADR and decreased 100-fold the IC50 from 8.98 microM with free ADR to 9.52 x 10(-2) microM. ADR-Fe-Tf was 10-fold more inhibitory to MCF-7/ADR cells than free ADR. Compared to Ga-Tf, ADR-Ga-Tf was 500- and 3000 fold more inhibitory to MCF-7 and MCF-7/ADR cells, respectively. These results demonstrated that ADR-Ga-Tf reverses resistance to free ADR and Ga-Tf in MCF 7/ADR cells. The distribution of ADR in both cell lines was examined by fluorescence microscopy. It was noted that ADR mainly accumulated in the cytoplasm around the nucleus in MCF-7/ADR cells, but in both the cytoplasm and nucleus of MCF-7 cells. However the conjugate of ADR-Ga-Tf allowed ADR to accumulate in the cytoplasm and nucleus of both the MCF-7/ADR and MCF-7 cells. Further investigation of MDR1 and MRP genes expression by RT-PCR demonstrated that Ga-Tf decreased expression of the MRP more than the MDR1 gene. Therefore the reversal of resistance to ADR by the ADR-Ga-Tf conjugate is mediated by the transferrin receptor transmembrane transport mechanism, redistribution of ADR into the nucleus of ADR resistant MCF-7/ADR cells and inhibition of MRP gene expression. PMID- 10810358 TI - Growth inhibition of Ph+ progenitor cells from CML patients using the tyrosine kinase inhibitor CGP57148B. AB - BACKGROUND: Different methods have been investigated for their purging capacity of contaminating CML cells in autologous stem cell products. CGP57148B, a tyrphostin, has been shown to be efficient in the reduction of cell proliferation of CML cell lines and primary CML cells, as well as in the inhibition of bcr/abl related tumor formation in animal models. MATERIALS AND METHODS: The effect of CGP57148B on purified CD34+ progenitor cells from BM, PB, or leukapheresis products of 8 CML patients was studied under serum-free cytokine-supplemented ex vivo culture conditions. RESULTS: FISH as well as RT-PCR analysis showed a significant reduction of Ph+ cells after 7 days ex vivo-culture in the presence of the tyrphostin. Growth of Ph- cells was almost unaffected by treatment with CGP57148B. CONCLUSION: Our results support the observation that CGP57148B can selectively inhibit proliferation of Ph+/bcr/abl+ primary CML cells under serum free cytokine-supplemented culture conditions. PMID- 10810359 TI - Androgen receptor gene mutations do not occur in ovarian cancer. AB - Genetic alterations have been frequently found in ovarian cancer. There is some indirect evidence indicating that mutation of the steroid receptor genes may play a role in the carcinogenesis of ovarian cancer. Human androgen receptor (hAR) gene mutations have been found in up to 50% of hormone-relapsed prostate cancer. The role of hAR mutation and its association with decreased expression in ovarian cancer has never been elucidated. In this study mutations of hAR gene in 38 human ovarian cancer cell lines with different AR expression pattern were studied using SSCP. No mutation of the hAR gene was found. Mutation of hAR gene is an infrequent event and therefore unlikely to be involved in the development of ovarian cancer. The decreased expression of hAR in advanced ovarian tumor is not due to genetic aberration of hAR. Mutation screening of hAR may not provide any information for risk assessment of developing ovarian cancer. PMID- 10810360 TI - Dietary phytosterol inhibits the growth and metastasis of MDA-MB-231 human breast cancer cells grown in SCID mice. AB - The objective of the present study was to investigate the effect of dietary phytosterols on the growth and metastasis of the human breast cancer MDA-MB-231 cell line xenografted in SCID mice. Two groups of animals were fed AIN-93G diet supplemented with 0.2% cholic acid and 2% sterol (cholesterol or phytosterol mixture) for 15 days before inoculation of the tumor into the right inguinal mammary fat pad. Tumor growth and food consumption were recorded weekly throughout the 8 weeks of the experiment. At the end of the experiment, the animals fed phytosterol had a 40% lower serum cholesterol and 20 and 30 fold higher serum beta-sitosterol and campesterol, respectively as compared to those fed cholesterol. There was no difference between the two groups in body weight and food consumption. However, the tumor size in animals fed phytosterols was 33% smaller (P < 0.03) and had 20% fewer metastases to lymph nodes and lungs than the cholesterol group. At termination, the tumor weight of the animals fed the phytosterol diet was also less (P < 0.07) than that of the cholesterol group. It is concluded that dietary phytosterols retard the growth and spread of breast cancer cells. PMID- 10810361 TI - Different expression of CD44, ICAM-1, and HSP60 on primary tumor and metastases of a human pancreatic carcinoma growing in scid mice. AB - Surface adhesion molecules play an important, but still not completely clarified, role in tumor metastasization. In this research, FACS analysis was employed to analyze surface expression of CD44H, CD44v5, CD44v6, ICAM-1 and HSP60 in human pancreatic adenocarcinoma cells growing in vitro or collected ex vivo from primary tumors and lung metastases of tumor-engrafted SCID mice. It was found that, in metastatic cells, the standard form of CD44 (CD44H) is down,-regulated, while a large fraction of cells express on membrane the splice variants v5/v6 and, in addition, ICAM-1 and HSP60. It was also apparent that two cell populations are present in lung metastases: a CD44neg population, including cells expressing CD44v5/v6, ICAM-1 and HSP60 and a population of CD44pos, CD44v5/v6neg, ICAM-1neg and HSP60neg cells. These results demonstrate that, in pancreatic adenocarcinomas, metastasization is correlated with expression of the CD44 variants v5 and v6. Moreover, this is the first report demonstrating HSP60 surface expression on metastatic cells. PMID- 10810362 TI - Lignin reduces ofloxacin-induced mutagenicity in Euglena assay. AB - The possible protective effect of sulphur-free beech lignin polymer on the mutagenicity of ofloxacin in Euglena gracilis was studied. The generation of oxygen species by ofloxacin and their possible interaction with lignin was verified by physico-chemical measurements. The UV absorbance spectra of ofloxacin with and without lignin showed no interaction between these two compounds. The production of superoxide anion radical (O2-) by ofloxacin was significantly reduced in the presence of lignin (AIR = 0.57 +/- 0.03, p < 0.01). Lignin, at concentrations of 125 and 250 micrograms/ml decreased the E. gracilis bleaching activity of ofloxacin to 39.9% and 2.8%, respectively. A lignin concentration of 500 micrograms/ml eliminated the bleaching activity of ofloxacin very efficiently. Our results are consistent with the concept that lignin biopolymer has the capability of reducing genotoxic activity by scavenging reactive oxygen species. PMID- 10810363 TI - Enhancement of drug-induced apoptosis by antisense oligodeoxynucleotides targeted against Mdm2 and p21WAF1/CIP1. AB - The p53 tumor suppressor gene plays an important role in DNA damage-induced apoptosis and, in general, inactivation of p53 contributes to poor response to chemotherapy. Apoptotic activity of p53 may be negatively modulated by expression of its downstream mediators, including Mdm2 and p21WAF1/CIP1. Consequently, these cellular pathways also represent potential targets for cancer therapy. This study investigated the effect of antisense oligodeoxynucleotides (ODNs), targeted against Mdm2 and p21WAF1/CIP1 on drug-mediated cell killing. Exposure of U2-OS osteosarcoma cells to DNA damaging agents, cisplatin or mitomycin C, caused upregulated expression of Mdm2 and p21WAF1/CIP1. Transient transfection of cells with antisense ODNs to Mdm2 mRNA inhibited Mdm2 protein expression and markedly enhanced apoptotic cell death induced by these drugs. Moreover, when p21WAF1/CIP1 expression was blocked by antisense transfection, drug-mediated cell killing was further accelerated. These results suggest that the enhanced expression of Mdm2 and p21WAF1/CIP1 may inhibit p53-mediated apoptosis and render cells resistant to the effects of DNA damaging agents. Consequently, antisense ODNs targeted against Mdm2 and p21WAF1/CIP1 could be employed in a potential therapeutic strategy sensitizing tumor cells to certain antineoplastic agents. PMID- 10810364 TI - Lipid peroxidation is associated with the inhibitory action of all-trans-retinoic acid on mammary cell transformation. AB - BACKGROUND: Retinoids are effective in reducing transformation of various cell types; however the role of lipid peroxidation has not been studied in this regard. MATERIALS AND METHODS: Retinoic acid (RA) and retinol were tested on several SV-40 large-T-antigen transformed bovine mammary fibroblast (MFB) lines that form foci on plastic with long-term culture. RESULTS: Dose response studies revealed that RA at 10(-6) M was the most potent dose in delaying the onset of foci formation and reducing total foci number. RA was always more effective than retinol. Addition of RA (10(-6) M) to MFB cells increased lipid peroxide (LPO) concentrations by approximately three-fold relative to untreated MFB cells or to RA or control treated normal mammary fibroblasts. The lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), acted synergistically with RA to increase LPO and cell death in MFB cells. The combination of the cyclooxygenase inhibitor, indomethacin, at 10(-4) M with 10(-6) M RA lowered MFB fibroblast cell numbers when compared to fibroblasts cultured singly with either RA or indomethacin. CONCLUSIONS: These data indicate that an increase in lipid peroxidation occurs specifically in tumor cells treated with RA and this may play a role in RA mediated suppression of cellular transformation in the mammary gland. Additionally, eicosanoid inhibitors may have an additive or synergistic effect with RA on the inhibition of mammary tumor cell transformation and proliferation. PMID- 10810365 TI - P27KiP1 overexpression inhibits the growth and doxorubicin sensitivity of HT29 human colon cancer cells in vivo. AB - We have previously shown that p27KiP1 plays a role in the tumor cell resistance of HT29 confluent monolayers to cytotoxic drugs in vitro. To determine whether p27KiP1 was a resistance factor to cytotoxic drugs in vivo we tested the effect of doxorubicin on p27KiP1-overexpressing HT29 tumors in nude mice. In this study we show that ectopic overexpression of p27KiP1 in HT29 human colon cancer cells decreases their tumorigenicity in vivo in nude mice. This decreased tumor growth was associated with increased p27KiP1 protein expression, studied by Western blotting in tumor extracts. Interestingly, the overexpressing-p27KiP1 tumors were significantly more resistant to intraveneous doxorubicin treatment than the control tumors. These results indicate that p27KiP1, which delays tumor growth could also increase tumor resistance to cytotoxic drugs in vivo. PMID- 10810366 TI - Bovine seminal ribonuclease exerts selective cytotoxicity toward neuroblastoma cells both sensitive and resistant to chemotherapeutic drugs. AB - BACKGROUND: Bovine seminal ribonuclease (BS-RNase) exerts selective cytotoxicity toward different types of tumor cells. In the present study, we tested the effects of BS-RNase on cultured neuroblastoma (NB) cells resistant to chemotherapeutic agents. The selectivity of the antitumoral activity of BS-RNase was evaluated using cultures of CD34+ hematopoietic stem cells. MATERIALS AND METHODS: Human NB cell lines including IMR-32, UKF-NB-2 and UKF-NB-3 were selected for resistance against vincristine, doxorubicin or cisplatin by exposure to increasing concentrations of the respective drug. The cytotoxicity of the drugs to NB cells was evaluated using a clonogenic assay in a methylcellulose medium. Peripheral blood progenitor cells were obtained from adult healthy donors by positive selection using specific anti-CD34+ antibodies. The toxicity of BS RNase to CD34+ cells was assessed in the direct clonogenic assay using methylcellulose medium or in ex vivo expansion culture supplemented with hematopoietic growth factors. RESULTS: In the clonogenic assay it was shown that BS-RNase completely inhibits growth of both parental NB cells and their sublines resistant to chemotherapeutic drugs at concentrations (up to 50 micrograms/ml) which have no significant influence on the growth of colony-forming units, granulocyte macrophage and erythroid burst-forming units. Moreover, BS-RNase had no effect on the ex vivo expansion of total hematopoietic cells or of colony forming cells from CD34+ progenitors. CONCLUSIONS: BS-RNase is a highly efficient agent against NB cells resistant to chemotherapeutic drugs. The lack of toxicity to hematopoietic progenitor cells suggests that BS-RNase is also likely to have tolerable hematopoietic toxicity. PMID- 10810367 TI - Membrane associated antitumor effects of crocine-, ginsenoside- and cannabinoid derivates. AB - In the present work a systematic study was initiated with crocine, ginsenoside and cannabinoid derivatives on multidrug resistant mouse lymphoma cells, viral tumor antigen expression and some human leukocyte functions. Among saffron derivatives, crocin and picrocrocin, triglucosyl and diglucosyl crocetin were ineffective on the reversal of multidrug resistance of lymphoma cells. Ginsenoside increased drug accumulation and tumor antigen expression at 2.0-20.0 micrograms/mL. Some cannabinoid derivatives such as cannabinol, cannabispirol and cannabidiol increased drug accumulation, while cannabidiolic acid, delta-9-THC and tetrahydro-cannabidiolic acid reduced drug accumulation of the human mdr1 gene transfected mouse lymphoma cells. The reversal of multidrug resistance is the result of the inhibition of the efflux pump function in the tumor cells. Crocetin esters were less potent than crocin itself in the inhibition of EBV early antigen expression. However crocin and diglucosylcrocetin inhibited early tumor antigen expression of adenovirus infected cells, but triglucosylcrocetin was less effective at 0.01-1.0 microgram/mL. The crocin had no antiviral effect [on HSV-2 infected vero cells] up to 25 micrograms/mL concentration. Ginsenosides had a moderate inhibitory effect except ginsenoside Rb1 (was the less effective) on the drug efflux pump. Among the cannabinoid derivatives the cannabinol and cannabispirol increased drug accumulation, while cannabidiolic acid and delta-8 THC, delta-9-THC and tetrahydro-cannabinol reduced drug accumulation in multidrug resistant mouse lymphoma cells. It is interesting that ginsenosides had a chemical structure-dependent immunomodulating effect by enhancing the activity of NK-cells and ADCC activities. PMID- 10810368 TI - Radiosensitivity of human breast cancer cells transduced with wild-type p53 gene is influenced by the p53 status of parental cells. AB - Induction of apoptosis with chemotherapeutic agents or radiation in tumours is frequently related to the status of those p53 gene of the tumours. To examine whether forced expression of the wild-type p53 gene in tumour cells can modulate their susceptibility to radiation and anti-cancer agents, we retrovirally transduced two types of human breast cancer cell lines, which respectively harboured a mutated p53 gene (OCUB-M) or wild-type p53 gene (YMB-1), with the wild-type p53 gene. Transduced cells which consistently expressed the wild-type p53 gene (OCUB-M/p53, YMB-1/p53) proliferated at the same rate as control cells which were transduced with the beta-galactosidase gene (OCUB-M/lacz, YMB-1/lacz). However, sensitivity to radiation was increased in OCUB-M/p53 cells but not in YMB-1/p53 cells. In vitro chemosensitivity to DNA-damaging anticancer agents such as cyclophosphamide and 5-fluorouracil was not influenced by the transduction of the wild-type p53 gene in either cells. Expression of the wild-type p53 gene in p53-mutated human breast cancer cells can therefore increase their sensitivity to radiation but not their chemosensitivity. Therapeutic effects following by the transduction of the wild-type p53 gene were not observed in breast cancer cells already bearing the wild-type p53 gene. PMID- 10810369 TI - Simian virus 40: the link with human malignant mesothelioma is well established. AB - Mesotheliomas are malignancies of the pleural, pericardial, and peritoneal surfaces with a mean survival of less than 1 year from the time of diagnosis (1). While mesotheliomas were extremely rare in the first half of this century, the incidence of these tumors has increased enormously in the last several decades. Presently, 2-3 thousand people in the US develop and die of mesothelioma each year (1). It is estimated that approximately 80% of mesotheliomas develop in people with a history of occupational asbestos exposure or in individuals with family member(s) professionally exposed to asbestos that brought home fibers on their clothing (1). Although conventional wisdom dictates that asbestos is the most commonly associated "environmental" factor with mesothelioma, asbestos does not transform human mesothelioma cells in tissue culture (2). This suggests that additional carcinogens act in concert with asbestos to cause mesothelioma. Recent evidence indicated that Simian Virus 40 (SV40) preferentially causes mesotheliomas in hamsters, and that SV40 is present in up to 80% of human mesotheliomas in the US and in Europe (reviewed in ref. 3 and 4). PMID- 10810370 TI - Molecular characterization of SV40 DNA in multiple samples from a human mesothelioma. AB - BACKGROUND: Prolonged exposure to asbestos, a potent carcinogen, has been the generally accepted factor responsible for the development of human mesotheliomas. Recent reports documenting the detection of SV40 DNA in human mesotheliomas suggest the possibility that this known tumor virus may be an additional factor involved in the development of some tumors. METHODS: A detailed analysis was performed by polymerase chain reaction and DNA sequencing of the genetic characteristics of SV40 viral DNA detected in samples taken from multiple sites of a human mesothelioma. RESULTS: A single virus variant was detected within the tumor that encoded a novel variable region at the C-terminus of the large T antigen oncoprotein. The viral regulatory region was predominantly archetypal in sequence (lacking duplications of the enhancer), typical of natural isolates. CONCLUSIONS: These data confirm previous reports from several laboratories showing an association of SV40 DNA with human mesotheliomas and provide the first evidence of a novel virus variant present in separated regions of a mesothelioma. PMID- 10810371 TI - SV40, JC and BK expression in tissue, urine and blood samples from patients with malignant and nonmalignant pleural disease. AB - BACKGROUND: Polyomaviruses are expressed in both human tumors and immunodepressed patients. Malignant and nonmalignant pleural effusions create an environment that could favor the expression of opportunistic viral infections. We studied if SV40, JC, and BK viral DNA can be amplified from biopsies obtained from different pleural diseases. MATERIALS AND METHODS: DNA was extracted from mesotheliomas (MM), nonspecific inflammatory and tubercular pleural biopsies, blood and urinary sediments from patients with MM, and pleural effusion cytological specimens. SV40, JC and BK viral early regions were amplified by PCR and analyzed by Southern Blot hybridization with specific probes. RESULTS: SV40 was positive in 9/23 MM, 5/18 tubercular and 1/7 nonspecific inflammatory biopsies, and 5/12 pleural effusion cytological specimens. JC was positive in 2/23 MM and in 7/15 urinary sediments. All blood samples were negative and BK was also negative in all samples. CONCLUSIONS: Tissue specific factors, characteristic of MM and TB, may contribute to expression of SV40 in these diseases. PMID- 10810372 TI - Simian virus 40 is not a cofactor in the pathogenesis of environmentally induced malignant pleural mesothelioma in Turkey. AB - Malignant pleural mesothelioma (MPM) continues to be a public health problem in Turkey, where exposure to environmental asbestos and fibrous zeolite (erionite) is the main cause of the disease. However, less than 5% of exposed individuals develop the disease, and numerous cases of MPM are documented each year in which the patient has no known exposure to either of these minerals. Thus, additional unknown factors act independently or as co-carcinogens in the development of MPM. Simian Virus 40 (SV40) may act as a co-carcinogen with asbestos in the pathogenesis of occupationally induced MPM. To determine if SV40 plays a role in the development of MPM in Turkey, we used PCR analysis to investigate if SV40 DNA sequences were present in 29 mesothelioma specimens from patients previously exposed to asbestos or erionite. PCR analysis revealed that all 29 tissue specimens from our patients did not contain SV40 DNA. 15 specimens from patients suffering from tuberculosis pleuresy were also SV40 negative. One mesothelioma and one osteosarcoma from Italy tested positive for SV40. Our results indicate that inorganic fibers, asbestos, and erionite remain the only known causal factors of mesothelioma in Turkey. The absence of SV40 in Turkish specimens and its presence in Italian specimens may be related to the fact that SV40 contaminated vaccines were not administered in Turkey. PMID- 10810373 TI - SV40 can be reproducibly detected in paraffin-embedded mesothelioma samples. AB - We studied tissue sections from 18 paraffin embedded mesothelioma specimens diagnosed by the Pathology Department of S. Chiara Hospital of Pisa. Using PCR analysis and Southern blot hybridization we examined the specimens for the DNA regulatory region of the virus. 10/18 (55.5%) of the samples tested contained SV40 DNA regulatory sequences, and of these positive samples, 80% were found to contain Tag sequences by PCR and Southern Blot hybridization. These results confirm that SV40 can be amplified and detected in paraffin embedded mesothelioma samples. PMID- 10810374 TI - Correlation of cisplatin sensitivity with differential alteration of EGFR expression in head and neck cancer cells. AB - Overexpression of human epidermal growth factor (EGFR) has been associated with a variety of human malignancies. The exact role of EGFR in human malignancies and its correlation with chemotherapeutiveness response has not been determined. Using a quantitative RT-PCR method, we previously studied the effects of cisplatin treatment on levels of EGFR mRNA in human papillomavirus (HPV)-positive head and neck cancer cell lines. In this report we extended these studies to HPV negative head and neck cancer cells. We also compared the growth inhibition and 50% inhibitory concentration (IC50) of cisplatin between these cells. We found that three of four HPV-negative cell lines had 3 to 5 times higher cisplatin IC50 values as compared to two HPV-positive cell lines. EGFR mRNA levels were increased after exposure to cisplatin in the cell lines with the higher IC50 values, while EGFR levels were reduced after cisplatin exposure in the cell lines with the lower IC50 values. These results suggest that the cisplatin sensitivity of head and neck cancer cells corresponds to subsequent alteration of EGFR levels following cisplatin treatment. PMID- 10810375 TI - The hollow fiber assay: continued characterization with novel approaches. AB - The hollow fiber assay, a unique in vivo model, permits the simultaneous evaluation of compound efficacy against multiple cell lines in two physiological compartments. This assay has been used to characterize in vivo activity of cytotoxic compounds. The purpose of the present study was to characterize and optimize this assay for compounds with a defined mechanism of action, specifically cell cycle inhibition. Two human tumor cell lines and one normal human cell line were loaded into polyvinylidene fluoride hollow fibers at two or more cell concentrations and grown in mice for 3-10 days. The data demonstrate the importance of characterizing the initial loading density of various cell lines in the evaluation of compounds. All studies were performed with cells in the linear part of the cell growth curves. Initial loading densities of 1-2 x 10(4) cells/fiber gave the greatest opportunity for growth in the three human cell lines tested (HCT116 colon carcinoma, NCI-H460 non-small cell carcinoma, and AG 1523 normal fibroblast). Utilizing the MTT assay, standard curves were constructed to correlate the final number of cells with optical density (OD) readings at 540 nm in order to calculate cell numbers in the fibers. Insights into the mechanism of action of cisplatin have been gained using Western blot analysis of the cell cycle markers PCNA (a protein present throughout the cell cycle) and Rb (a protein that acts as a tumor suppressor gene product) from the hollow fiber cells. In cisplatin-treated NCI-H460 cells both PCNA and Rb phosphorylation decreased, suggesting the arrest of the cells prior to the S phase. Standard therapeutic agents, cisplatin, racemic flavopiridol, cyclophosphamide and mitomycin C, were evaluated independently in the hollow fiber assay and the xenograft model. The data demonstrate that compounds active in the hollow fiber assay are also active in the xenograft. PMID- 10810376 TI - Metastatic potential of human uroepithelial cancer cells is not dependent on their adhesion to E-selectin. AB - In our previous studies we have found that human uroepithelial cell lines differ in their expression of sialosyl LewisA antigen. We have also shown that among the studied cell lines, only Hu 1703He cells with the highest expression of this tetrasaccharide bind to E-selectin-expressing cells. In the present study we put forward the question, of whether sialosyl LewisA-mediated adhesion of uroepithelial cells to E-selectin is important in the formation of metastases. The HCV 29T and Hu 1703He cells, representing two uroepithelial cell lines, were transplanted into NCr nu/nu mice. Hu 1703He cells express on their surface a high level of sialosyl LeA antigen, while HCV 29T cells are sialosyl LewisA-negative. We have shown that human uroepithelial cancer cells, in addition to their tumorigenic and invasive properties, are highly metastatic when inoculated into athymic nu/nu mice. The ability to form secondary tumour foci in lung and liver seems to be independent of the expression of sialosyl LewisA antigen. PMID- 10810377 TI - The expression of cadherins in human neuroblastoma cell lines and clinical tumors. AB - Cadherins are Ca(2+)-dependent cell-cell adhesion molecules which play crucial roles in the cell-cell interactions during development, tumorigenesis and metastasis. The absence of N (neural)-cadherin is correlated with the onset of neural crest migration and its reappearance is correlated with the cessation of migration and precedes gangliogenesis. We investigated the expression of cadherins including N-cadherin in five cell lines and eleven clinical specimens of human neuroblastomas, which originated from neural crest cells. We found that three of the neuroblastoma cell lines and all the clinical specimens were positive for the expression of the N-cadherin protein. The other two neuroblastoma cell lines were negative for the expression suggesting they originated from migrating neural crest cells. All these cell lines and clinical samples expressed either cadherin-6, cadherin-11 or both, i.e. cadherins expressed on neural crest cells, supporting their neural crest origin. PMID- 10810378 TI - Radioimmunoscintigraphy and radioimmunotherapy in cancer: principles and application. AB - Radioimmunoscintigraphy (RIS) and radioimmunotherapy (RIT) are recent approaches in the diagnosis and treatment of cancer. They take advantage of the antibody specificity of tumor surface antigens and of the emitted radiation from suitable radioisotopes, as a means of imaging (RIS) or therapy (RIT). Research into RIS and RIT radiolabelled agents remains an ongoing process. Principles governing the choice of radionuclides, labelling protocols, antibody suitability, and optimization of "tumor to normal tissue ratios" are the same for both RIS and RIT. The investigational stages of the labelled product, prior to clinical application, are also the same. These stages include radiochemical and radiobiological evaluation as well as determination of immunoreactivity. Furthermore, RIS may be considered as the first stage in development, before progressing on to RIT. Differences between RIS and RIT are associated with the application of each technique, that is, the type of radiation emitted by the isotope, dosage regimens, haematopoetic toxicity and the appearance of human antimurine antibody response (HAMA). RIS has found widespread clinical application, detecting a variety of tumors. However, its potential lies in patient management and in detecting metastases. On the other hand RIT is still in its infancy. It appears promising, and for the moment is used as a complementary technique to surgery and/or chemotherapy in clinical trials on cancer treatment. Finally, incorporation of these basic principles arising from past experiences, into the design of RIT trials improve responses. PMID- 10810379 TI - Reduced expression of MHC class I antigen in human cancer cell lines with defective LMP-7. AB - The function of major histocompatibility complex (MHC) class I molecule is to sample peptides derived from intracellular proteins and to present these peptides to CD8+ T cells. We analyzed twenty-nine human cancer cell lines for expression of the TAP-1/2 and LMP-2/7 genes and identified whether defects in these genes were associated with the level of expression of MHC molecules in cancer cells. Expression of TAP-1 was positive in 86% (25/29) of the cell lines, TAP-2 expression was positive in 82% (24/29), LPM2 was positive in 86% (25/29), and LMP 7 was positive in 52% (15/29) at the cell lines. In fifteen cell lines that expressed LMP-7, the mean fluorescence intensity (MFI) was 825.1 +/- 123.2; however, in the cell lines that lacked LMP-7, the MFI was 314.7 +/- 77.2. These data suggest that the expression level of MHC class I molecules is associated, in most cases, with the loss of the LMP-7 gene but without the loss of the TAP-1, TAP-2 or LMP-2 genes. PMID- 10810380 TI - Inhibitory effects of gallium chloride and tris (8-quinolinolato) gallium III on A549 human malignant cell line. AB - The effects of two gallium (Ga) compounds, Ga chloride (GaCl3) and tris(8 quinolinolato)Ga (III) on the viability of A549 human malignant lung adenocarcinoma cells were investigated. The results demonstrated that both drugs reduced the viability of A549 cells but to different extents. The inhibitory effects of tris(8-quinolinolato)Ga (III) were 10 times more profound than those produced by GaCl3. The IC50 was obtained with 2.5 microM of tris(8 quinolinolato)Ga (III) and 25 microM GaCl3 after an exposure time of 48 hours. Further, whereas the inhibitory effects of GaCl3 were both dose and time dependent those of tris(8-quinolinolato)Ga (III) appeared to be only dose dependent, indicating differences in their mechanism of action. Comparison with data drawn from the literature suggests that GaCl3 seems to be in the same range of activity as Ga nitrate or Ga-pyridoxal isocotinoyl hydrazone. Tris(8 quinolinolato)Ga (III) could be as effective as transferrin-Ga, but with the advantage of oral administration and a greater bioavailability of the tris(8 quinolinolato)Ga (III) compound. PMID- 10810381 TI - In vitro evaluation of mutant HSV-1 thymidine kinases for suicide gene therapy. AB - Herpes Simplex Virus type 1 (HSV-1) thymidine kinase (TK) is currently the most widely used suicide agent for gene therapy of cancer. Tumor cells that express HSV-1 thymidine kinase are rendered sensitive to prodrugs due to preferential phosphorylation by this enzyme. While ganciclovir (GCV) is the prodrug of choice for use with TK, this approach is limited in part by the toxicity of this prodrug. From a random mutagenesis library of over a million mutant thymidine kinases, ten thymidine kinase variants were identified on the basis of activity towards ganciclovir and acyclovir (Black ME, Newcomb TG, Wilson H-MP and Loeb LA: Proc. Natl. Acad. Sci. U.S.A. 93: 3525-3529, 1996). Six mutants described here contain three to six amino acid changes and render mammalian cells more sensitive to acyclovir (ACV) including one that demonstrates an 8.5-fold reduction in IC50 compared to wild-type TK. These novel enzymes could provide benefit to ablative gene therapy by now making it feasible to use the relatively non-toxic acyclovir at nanomolar concentrations. PMID- 10810382 TI - Polyploidization induced by acridine orange in mouse osteosarcoma cells. AB - This study was undertaken to clarify the in vitro effect of acridine orange (AO) on the cell kinetics of mouse osteosarcoma cells, as well as the mechanism of cell growth inhibition induced by AO. A mouse osteosarcoma cell line (MOS), established from a radiation-induced mouse osteosarcoma, was cultured under exposure to 0.05, 0.5, 5, and 50 micrograms/ml of AO, either continuously or for 10 minutes. The cell kinetic analysis was performed using the following parameters: tumor cell growth by trypan blue exclusion test, mitotic activity, DNA synthetic activity by BrdU labeling and DNA ploidy by cytofluorometry. The results showed that continuous exposure to 5 and 50 micrograms/ml of AO or 10 minute exposure to 50 micrograms/ml of AO quickly killed the tumor cells within 12 hours, whereas continuous exposure to 0.5 microgram/ml of AO or 10 minute exposure to 5 micrograms/ml of AO gradually inhibited tumor cell growth. Under the latter conditions, mitotic activity was rapidly and completely inhibited within 48 hours but DNA synthetic activity was not completely inhibited even after 96 hours. DNA ploidy analysis demonstrated that most of the tumor cells arrested at the S-G2 phase after 12 hours, followed by G2 phase arrest after 24 hours and progressive DNA synthesis to a higher DNA ploidy class after 48 to 96 hours. We therefore concluded that a high concentration of AO has a strong cytocidal effect due to cytotoxicity whilst a moderate concentration of AO induces progressive and synchronous polyploidization by mitotic inhibition without DNA damage in MOS cells. We presume that this in vitro effect on MOS cells may be caused by protein synthetic inhibition after transfer RNA inactivation caused by AO binding. PMID- 10810383 TI - Intracellular binding sites of acridine orange in living osteosarcoma cells. AB - There have been many reports concerning the intracellular binding sites of acridine orange (AO), although the actual localization of AO in living cells remains controversial. This study was undertaken to clarify the intracellular localization of AO in living mouse osteosarcoma cells by cytochemical staining. A mouse osteosarcoma cell line (MOS) was cultured and continuously exposed to 0.5 microgram/ml of AO. The intracellular localization and stainability of AO the living tumor cells was morphologically detected by a high resolution fluorescence microscope. To detect the intracellular microstructure, cytochemical staining with rhodamin 123 for mitochondria, acid phosphatase for lysosome, Sudan-black for fat vesicle and toluidine blue for glucosaminoglycan were performed using fixed cells. The results showed that both the nucleus and cytoplasm of tumor cells at 10 minutes after exposure to 0.5 microgram/ml of AO emitted green fluorescence, which was especially intense in the nucleolus, but not brilliant in the nucleus and was granular orange to red fluorescence in the perinuclear particles. This stainability of AO was different from that of rhodamin 123, Sudan black or toluidine blue, but similar to that of acid phosphatase. Based on these results, we conclude that the green fluorescence may have derived from AO binding to double stranded RNA, not to DNA, and that orange fluorescence may have derived from aggregated AO binding to lysosome. PMID- 10810384 TI - Cytotoxicity in vitro and preliminary antitumor activity in vivo of a novel organotin compound. AB - The cytotoxic effect and antitumor activity induced by the novel organotin compound triethyltin(IV)lupinyisulfide hydrochloride, have been investigated. Different patterns of antiproliferative effects have been observed in a panel of human tumor cell lines in vitro. Toxicity studies in mice reported acute toxicity at the doses of 21 and 17.5 mg/kg which progressively disappeared at lower concentrations. On this basis, the doses of 3.5, 7 and 14 mg/kg were selected to assess the antitumor activity in vivo against the P388 leukemic cells xenografted in mice. This compound was able to induce a dose-dependent significant reduction of tumor volume, up to 46%, at the highest concentration (p = 0.0062) without important toxicity, as also confirmed by histological analysis of the main organ tissues. This preliminary study seems to hold interest for further investigations in different tumor models as well as for the evaluation of optimal drug route and schedule. PMID- 10810385 TI - The normal epithelial cell-specific 1 (NES1) gene is up-regulated by steroid hormones in the breast carcinoma cell line BT-474. AB - The normal epithelial cell-specific 1 (NES1) gene encodes a serine protease which was found to be down-regulated in breast cancer. There is evidence that NES1 acts as a tumor suppressor gene in breast cancer cells. To further understand its role in breast tumorigenesis, we investigated the effect of estrogens, androgens, and progestins on NES1 gene expression, in the breast cancer cell line BT-474, at the transcription level. The reverse transcriptase polymerase chain reaction method was used to monitor changes in the NES1 mRNA. Our experiments showed that NES1 gene expression is up-regulated promptly in response to 17 beta-estradiol, 5 alpha-dihydrotestosterone (DHT) and norgestrel stimulation. NES1 gene mRNA started to increase 2 hours after estradiol stimulation and 8 hours after DHT stimulation. The stimulation of NES1 by estradiol can be dramatically blocked by the estrogen antagonists ICI 182,780 and 4-hydroxytamoxifen. Mifepristone (a synthetic antiprogestin) can partially block the up-regulation of the NES1 gene by norgestrel. Dose-response experiments indicated that the lowest stimulatory concentration of 17 beta-estradiol, DHT, and norgestrel is 10(-11) M, 10(-10) M, and 10(-10) M, respectively. The production of NES1 mRNA increased coordinately with increasing concentration of the stimulants. These results suggest that the NES1 gene is primarily regulated by estrogen, but also by androgen and progestin in the breast cancer cell line BT-474. It appears that NES1 may be involved in a pathway that counter balances the action of estrogens and androgens in steroid hormone responsive tissues. PMID- 10810386 TI - Multiwavelength videomicrofluorometric study of cytotoxic effects of a marine peptide, didemnin B, on normal and MDR resistant CCRF-CEM cell lines. AB - The development of multidrug resistance (MDR) in heterogeneous cell sensitive and resistant populations to a variety of clinically important cytotoxic drugs poses a major obstacle to cancer chemotherapy. Didemnin B, a marine cyclic depsipeptide, displays interesting biological properties: antiviral activity, inhibition of DNA, RNA and protein synthesis, initiation of apoptosis and ability to block the cell cycle. As very little is known about its mode of action, we studied the effect of increasing doses of Didemnin B on sensitive and resistant human leukemic lymphoblast cell lines. The fluorescence of living cells simultaneously stained with Hoechst 33,342, Rhodamine 123 and Nile Red, were analyzed in a multiparametric approach involving multiwavelength microfluorometry. High concentrations of Didemnin B induced, in the sensitive cell line, a very early decrease in the energetic state of the mitochondria that occurs before a significant decrease of nuclear DNA content, observed simultaneously on sensitive and resistant cells, that could be related to an apoptosis process. Furthermore low Didemnin doses (50 nM) affected CEM-WT and CEM VLB differently, while higher doses (200 nM-250 nM and over) affected the two cell lines in the same way. This indicated that, at these doses, the membranar Pgp has no effect on the mode of action of Didemnin, suggesting that Didemnin does not need to be internalized to be active. PMID- 10810387 TI - c-erbB-2 oncoprotein is overexpressed in poorly vascularised squamous cell carcinomas of the head and neck, but is not associated with response to cytotoxic therapy or survival. AB - The prognostic and predictive role of c-erbB-2 oncoprotein in human malignancies remains controversial. The pattern of expression and the biological behaviour of c-erbB-2 protein was investigated in a series of 89 locally advanced squamous carcinomas of the head and neck. Possible associations between c-erbB-2 and T,N stage, histological grade, microvessel density and the expression of bcl-2 and p53 proteins were sought. Biopsy material had been collected prior to the initiation of treatment which consisted of platinum based induction chemotherapy or concurrent chemotherapy and radiotherapy. In all cases, follow-up of at least 2 years duration was available. Positive staining was seen in 27 out of 89 (30.4%) cases and was invariably cytoplasmic, exhibiting strong reactivity in more than 50% of tumour cells. Such c-erbB-2 overexpressing tumours were not associated with a response to radiotherapy and/or induction chemotherapy. Further, survival analysis did not disclose any difference in the overall or relapse free survival between c-erbB-2 positive and negative cases. Similarly, no relationship was found with T,N-stage, histological grade and bcl-2 or p53 expression. There was, however, a significant inverse association between c-erbB 2 expression and microvessel density (p = 0.0001). Interestingly, tumours with low vascular grade (VG; MS < 26) and positive c-erbB-2 expression were less frequently associated with local aggressiveness compared to c-erbB-2 negative tumours and low VG (5/27 vs. 12/18; p = 0.001), or to c-erbB-2 negative tumours and high VG (MS > 27) (5/27 vs. 19/44; p = 0.03). These differences were statistically significant but were not related to treatment response or prognosis. Nonetheless, when patients with highly angiogenic tumours were analysed for survival, irrespective of the c-erbB-2 status, they showed a poorer prognosis than those having a low microvessel density (p = 0.06 and 0.05 for overall and relapse free survival, respectively). Six out of 89 patients presented with distant metastases, five of which had tumours negative for c-erbB 2 expression. Our results seem to indicate that the pattern of c-erbB-2 expression in locally advanced inoperable head and neck cancer is exclusively cytoplasmic. c-erbB-2 reactivity is of little value in predicting survival or chemo-radiotherapeutic response. Expression of genes related to angiogenesis or other invasion and migration pathways are, apparently, more important. PMID- 10810388 TI - In vitro effects of free 211At,211At-albumin and 211At-monoclonal antibody compared to external photon irradiation on two human cancer cell lines. AB - BACKGROUND: The aim of this study was to perform various 211At irradiations of importance for the evaluation of 211At-radioimmunotherapy, and compare the effect with that of low linear energy transfer (LET) radiation. MATERIALS AND METHODS: All irradiations were performed on low-concentration single-cell suspensions. Growth assays using 96-well plates were used to estimate apparent cell survival. Centrifuge tube filters were used to estimate the cell uptake and binding of 211At. RESULTS: A relative biological effect (RBE) of 12 +/- 2 (Colo-205) and 5.3 +/- 0.7 (OVCAR-3) was found from 211At-albumin irradiations. There was a 174 +/- 28 times higher free 211At concentration in the cell fraction than in the surrounding medium. For 211At-MAb, an 8,000-30,000 times higher concentration in the cell fraction was achieved, compared to the medium. Corrected for the uptake, an average of 31 +/- 2 ([211At]-astatine) or 26 +/- 5 ([211At]-MAb) decays per cell were required for 37% survival of Colo-205 cells. An average of 19 +/- 3 decays ([211At]-astatine) were required per OVCAR-3 cell. CONCLUSIONS: Cell uptake and binding of 211At was unexpectedly high, possibly favouring its therapeutic use. The binding is probably to the cell surface. The RBE is 5.3 +/- 0.7 for OVCAR-3 and 12 +/- 2 for Colo-205 cells. PMID- 10810389 TI - The intestinal absorption of camptothecin, a highly lipophilic drug, across Caco 2 cells is mediated by active transporter(s). AB - The intestinal transport properties and kinetics of camptothecin (CPT) through Caco-2 cell monolayers were characterized by determining concentration-, temperature-, and ATP-dependence as well as the effect of selected inhibitors. The absorptive permeability (Peff) of CPT was found to be concentration dependent and saturable (K(m) = 31.2 +/- 6.9 microM) indicating the involvement of a high affinity, low capacity transport system. CPT transport was also temperature dependent and inhibited by sodium azide plus 2-deoxyglucose, which deplete cellular ATP, further suggesting that an active, carrier-mediated transport system contributes to CPT absorption. Based on inhibition studies, the involvement of organic anion and cation transporters was implicated but not conclusively demonstrated. Total CPT transport decreased four fold with increasing pH from 5.5 to 8.5 indicating that CPT lactone contributed more significantly to overall CPT transport than CPT carboxylate. The results of these studies suggest that CPT absorption is mediated by multiple mechanisms including significant passive diffusion and active transport components. Since typical substrates for intestinal carriers are hydrophilic and charged, the involvement of putative absorptive carriers in the transport of CPT is a novel finding that may give insight into the erratic oral bioavailability of CPTs observed in the clinic. PMID- 10810390 TI - Promoter specific sensitivity to inhibition of histone deacetylases: implications for hormonal gene control, cellular differentiation and cancer. AB - Alterations in histone acetylation status appear to play a central role in the regulation of neoplasia, tumor suppression, cell cycle control, hormone responsiveness and senescence. These alterations of chromatin control gene transcription. The histone acetylation status is regulated by the equilibrium of histone acetyl-transferase activity (HAT) and the histone deacetylase activity (HDAC). Commonly, DNA-transfection assays are used to measure the effect of histone acetylation and deacetylation on gene transcription. Here we have analyzed the response of various viral long terminal repeats and vertebrate promoters to the specific histone deacetylase inhibitor trichostatin A (TSA). We show that the activity of many, but not all, promoters is increased upon TSA treatment. Interestingly, the lysozyme promoter exhibited TSA resistance, while the activity of metallothionine, the human growth hormone, and the thymidine kinase promoters was increased. Furthermore, we found that all tested viral promoters are induced by TSA. Analysis of the transcriptional behaviour of the thyroid hormone receptor (TR), the cellular homologue of the v-erbA oncogene, revealed that TSA reduced the gene silencing function but had no influence on the hormone-induced gene activation function of the receptor. These results on gene specific effects, together with the HDAC structural data (1), may be a basis for the development of HDAC inhibitors as antitumor agents. PMID- 10810391 TI - Influence of various administration routes on the antitumor efficacy ofTT-232, a novel somatostatin analog. AB - TT-232 a novel tumor-selective somatostatin analog with a five residue ring structure (D-Phe-Cys-Tyr-D-Trp-Lys-Cys-Thr-NH2) was developed by us and published in an earlier work. This synthetic heptapeptide had no effect on growth hormone release, but had a remarkable tyrosine-kinase inhibitory effect and inducted apoptosis. The aim of this study was to compare the therapeutic efficacy of TT 232 used in various long-term administration routes and treatment schedules. The effectiveness of TT-232 was studied on different rodent tumors transplanted to inbred mice from SPF breeding. Intermittent treatment by injections and continuous infusion of TT-232 using a s.c., i.p. or i.v. implanted Alzet type osmotic minipump were compared for therapeutic efficacy. The treatments were started either on the day subsequent to tumor transplantation or after the development of a tumor. On the basis of survival and tumor growth inhibition the infusion of TT-232 for 14 days using an implantable osmotic pump proved to be a much more effective route of treatment in both s.c. and i.v. administration than the intermittent injections applied twice a day for 2 weeks. In the case of S-180 sarcoma the continuous administration of TT-232 for 14 days using s.c. implanted osmotic pump resulted in 60% the i.v. infusion produced 40% long-term (over 80 days) and tumor free survivors. By the continuous administration of TT-232, an 80 100% tumor growth inhibitory effect and a considerable retardation of tumor development could be achieved. Continuous infusion from implanted pumps ensured a constant drug level and resulted in a well-defined, consistent pattern of drug exposure over the full duration of drug administration. In our study the route of infusion has been shown to increase drug efficacy relative to conventional delivery methods. PMID- 10810392 TI - Oncocalyxones A and C, 1,4-anthracenediones from Auxemma oncocalyx: comparison with anticancer 1,9-anthracenediones. AB - Oncocalyxones A and C are 1,4-anthracenediones isolated from Auxemma oncocalyx (Boraginaceae) that have been shown to be cytotoxic to tumor cells in vitro. The present study compared the cytotoxicity of these compounds with that of two conventional anticancer agents doxorubicin and mitoxantrone, both 1,9 anthracenediones, in a panel of human tumor cell lines. The effect on cell growth was examined using an MTT microtiter assay in two leukemia lines, five solid tumor lines of different histological origin, and two multidrug-resistant sublines of a lung tumor line. The oncocalyxones showed much lower potency than the 1,9-anthracenediones, but were similarly more cytotoxic to leukemia cells compared to solid tumor lines. However, in the multidrug-resistant cells with 10 to 500 times decreased sensitivity to doxorubicin, the cytotoxicity of oncocalyxones A and C was only modestly reduced by about twofold, 1,4 Anthracenediones may be a promising novel class of chemotherapeutic agents effective against multidrug resistant tumors. PMID- 10810393 TI - Serum evaluation of P53 protein in patients with gynaecological cancer. AB - Sera from patients with gynaecological cancer including the ovary, endometrium or cervix were examined for p53 protein, using the Pantropic p53 quantitative ELISA. Patients with benign gynaecological pathologies were included as a control group. p53 values ranged from undetectable to high levels of the protein (range: 0-531 pg/ml). Using the value of 200 pg/ml as the cut-off, p53 serum levels were found to be elevated in 23% of the patients with ovarian cancer, in 16% of the patients with endometrial cancer and in 14% of the patients with cervical cancer. In the control group, increased serum p53 levels were found in 3.3% of patients. No differences were observed among the groups with different types of cancer or at different stages, but the differences between the cancer groups and the control group were statistically significant. Our results suggest that serum p53 evaluation could be an adjunctive tool to the diagnostic laboratory tests for preoperatively gynaecological cancers and both a competitive and alternative useful procedure for the detection of p53 gene mutations. PMID- 10810394 TI - Enhanced cellular radiation sensitivity of androgen-independent human prostate tumor cells by onconase. AB - The RNase-like onconase, isolated from amphibian oocytes, showed increases in median tumor pO2 in solid tumors (1). This led us to consider if onconase could decrease cellular O2 consumption (QO2) on 9L rat glioma as well as DU145 human prostate adenocarcinoma cells. Using a Clark-type electrode chamber, we observed that onconase significantly inhibited QO2 in both tumors we tested. Since onconase-induced reduction in QO2 could lead to increases in radiation sensitivity, due to the diffusion of O2 to previously hypoxic tumor cells, we used androgen-insensitive DU145 cells to study onconase-induced changes in radiation sensitivity in vitro. Radiation sensitization was achieved with > 5 micrograms/ml of onconase, regardless of the p53 status of tumor cells. Data presented here suggested that onconase-induced enhancement in radiation sensitization in vitro of androgen-insensitive prostate cancer cells warranted further studies of radiation responses in vivo, prior to clinical settings for the advanced-stages of prostate cancer. PMID- 10810395 TI - Anticancer effect of PMEDAP--monitoring of apoptosis. AB - Antitumor effect of N-9-[2-(phosphonomethoxy) ethyl]-2,6-diaminopurine (PMEDAP) was studied in an in vivo model of s.c. transplanted Sprague-Dawley (SD/cub) rat T-cell lymphomas. Three individual SD/cub neoplasias (SD10/96, SD14/97, SD1/90) of different phenotypes were used. During the treatment, survival of the rats, increase of lymphoma mass, and DNA fragmentation detected by APO/BRDU kit, as well as Bcl2 and p53 protein expression, were followed. The study gives evidence of the positive therapeutic effect of PMEDAP in two of the three tested lymphomas, SD10/96 and SD14/97. Slowly growing SD1/90 lymphoma differs from the others in a uniform karyotype with trisomy of chromosome 11, CD4- immunophenotype, heterogeneous cellular morphology and constitutive expression of p53 protein found in some neoplastic cells. Thus, the diverse anticancer efficacy of PMEDAP treatment among SD/cub lymphomas could be associated with the different phenotypes of individual neoplasias. PMID- 10810396 TI - Analysis of the immunoreactivity of three anti-p53 antibodies and estimation of the relations between p53 status and MDM2 protein expression in ovarian carcinomas. AB - BACKGROUND: The p53 growth suppressor is inactivated in human tumors by several distinct mechanisms, such as point mutations, binding to viral proteins and association with the MDM2 protein. Little is known about the expression of different immunologically distinct forms of p53 and MDM2 protein in human tumors and especially in ovarian carcinomas. MATERIALS AND METHODS: The overexpression of p53 and MDM2 oncoproteins was examined in a series of 46 ovarian carcinomas, taking into account the conventional pathological variables. The comparison of p53 and MDM2 expression in tissue sections and respective cyst and-or ascitic fluid cells was also performed. For the determination of the p53 expression the reactivity of three commonly used anti-p53 antibodies (DO7, PAb240, PAb1620), which detect immunologically distinct subclasses of p53, were analyzed in relation to the MDM2 status in individual patients. RESULTS: The detection of the p53 expression was clearly related to the antibody applied. DO7 antibody appears to be superior to both PAb240 and PAb1620 in immunohistochemical tests. Nuclear MDM2 protein overexpression was found in 17.4% of cases and usually it was associated with p53 accumulation. There was no significant correlation between p53 as well as MDM2 expression and histological subtypes, staging and grading parameters of carcinomas however p53 accumulation was detected more often in III/IV than in I/II FIGO stages. CONCLUSION: In ovarian carcinomas significant inter- and intratumoral heterogeneity in p53 and MDM2 expression was identified and different MDM2/p53 phenotypes were revealed. The restriction of MDM2 overexpression to a small subset of neoplasms indicates that this oncoprotein plays a minor role in ovarian carcinogenesis. PMID- 10810397 TI - Fhit expression is absent or reduced in a subset of primary head and neck cancer. AB - BACKGROUND: The inactivation of the FHIT gene at 3p14.2 by various mechanisms might be of importance in head and neck squamous cell carcinoma (HNSCC). Most reports are based on DNA and RNA findings of intragenic deletions and abnormal transcripts. MATERIAL AND METHODS: To study the protein expression of this putative tumour suppressor gene, we analysed 48 HNSCCs by immunohistochemistry using a polyclonal antibody (ZR44). The results were compared with mutation analysis, clinical data and loss of heterozygosity (LOH) data at 3p14.2. RESULTS: Complete absence of Fhit expression was detected in 8 out of 48 of tumours (17%) and 3 tumours (6%) showed heterogenous staining. The overall frequency of LOH for microsatellite D3S1234 was 64% and 5/7 of Fhit negative tumours exhibited LOH. CONCLUSION: Our findings provide further evidence that FHIT is inactivated in a subtype of HNSCC; however, the incidence of lack of Fhit expression compared to the high frequency of LOH on chromosome 3p supports the notion of additional tumour suppressor genes at 3p14. PMID- 10810398 TI - P-glycoprotein expression is a marker for chemotherapy resistance and prognosis in advanced ovarian cancer. AB - BACKGROUND: It was our aim to study the prevalence and possible prognostic and predictive significance of the expression of P-glycoprotein, a transmembrane transport protein related to classical multidrug resistance, in patients with advanced ovarian cancer. STUDY DESIGN: Tumor tissue from 73 previously untreated patients with FIGO stage 3 ovarian cancer was examined with immunohistochemistry for the expression of P-glycoprotein before and after chemotherapy. Response to 4 cycles of combination chemotherapy with cisplatin and epirubicin was assessed with second look laparotomy. The log rank test was used for univariate survival and the Cox proportional hazards regression model for multivariate survival analysis. RESULTS: P-glycoprotein expression was detected in 47% of untreated cases, and correlated with unfavourable prognostic factors such as advanced age, presence of ascites and larger residual disease deposits after primary surgery. P glycoprotein negative cases responded significantly better to chemotherapy (P < .001). In the multivariate survival analysis P-glycoprotein expression was an independent predictor of both overall (P = .045) and progression free (P = .006) survival. When P-glycoprotein expression and residual disease status were considered together, the patients could be divided in three clearly distinct prognostic groups (P = .0009). CONCLUSION: P-glycoprotein expression is a predictor of response and survival in a uniformly treated and followed cohort of advanced ovarian cancer patients. PMID- 10810399 TI - Multicenter randomized prospective study of adjuvant chemotherapy with UFT and mitomycin C in advanced colorectal cancer. AB - BACKGROUND: A randomized prospective trial was performed to determine the efficacy of preoperative and postoperative adjuvant oral UFT, administered with mitomycin C (MMC) after resection for advanced colorectal cancer. MATERIALS AND METHODS: A total of 126 patients were entered in the study. The patients received UFT (400 mg daily) administered orally for seven days prior to surgery and were randomly assigned to two groups immediately following surgery. Group A received MMC postoperatively; Group B received the same regimen as Group A, plus administration of UFT orally at a dose of 400 mg daily for one year. RESULTS: The survival results revealed no significant difference between groups A and B. In patients with nuclear DNA aneuploid tumors, the hematogenous recurrence rate after curative surgery was lower in Group B than in Group A (P = 0.0656). CONCLUSIONS: Preoperative and postoperative adjuvant oral UFT, administered with MMC after curative resection, may be effective in preventing hematogenous recurrence in colorectal cancer patients with nuclear DNA aneuploidy tumors. PMID- 10810401 TI - Limited value of preoperative serum analyses of matrix metalloproteinases (MMP-2, MMP-9) and tissue inhibitors of matrix metalloproteinases (TIMP-1, TIMP-2) in colorectal cancer. AB - PURPOSE: We studied whether preoperative serum levels of free MMP-2, the MMP 2/TIMP-2 complex, and total amounts of MMP-9, TIMP-1 and TIMP-2 correlated to the tumor stage and prognosis in colorectal cancer. METHODS: Samples from 158 patients operated on for colorectal cancer (100 colon, 58 rectum) and samples from 80 healthy blood donors were analyzed using an ELISA technique. One hundred and thirty-three patients were resected for cure, (31, 61, and 41 in Dukes' stages A, B, and C, respectively). At follow-up in January 1998, 44 patients had died from their cancer after a median time 14 months (range 2-55). Fifteen patients died without tumor relapse. Ninety-nine patients were alive after, a median time of 46 months (range 17-68). RESULTS: Wide, overlapping ranges were observed for all factors both in the patients and in the control group. The patients as compared to the control group had significantly higher levels of free MMP-2 and total amounts of MMP-9, TIMP-1 and TIMP-2, whereas the level of the MMP 2/TIMP-2 complex was significantly lower. TIMP-1 was significantly higher in Dukes' D compared to Dukes' A-C cases; the other factors did not correlate to tumor stage. Elevated TIMP-2 levels (median cut-off limit), only, correlated to worse prognosis when analysed in all patients (p < 0.05). None of the factors (median cut-off limit) correlated to survival in Dukes' A-C patients; analyses based on the upper quartile cut-off limit demonstrated that elevated MMP-2 levels correlated to shorter survival time (p < 0.05). CONCLUSION: Serum analyses of free MMP-2 the MMP-2/TIMP-2 complex and total amounts of MMP-9, TIMP-1 and TIMP-2 are of limited value for tumor staging and prognosis in colorectal cancer. PMID- 10810400 TI - Single agent docetaxel plus granulocyte-colony stimulating factor (G-CSF) in previously treated patients with advanced non small cell lung cancer. A phase II study and review of the literature. AB - The use of salvage chemotherapy in advanced non small cell lung cancer (NSCLC) is controversial. However, many patients need to be treated in order to achieve relief of their symptoms. Docetaxel (taxotere) is one of the most active drugs for the treatment of advanced NSCLC and several studies have also shown its effectiveness in pretreated patients. In the present study, 23 patients were treated in order to evaluate both the effectiveness and toxicity of the single agent docetaxel. Furthermore, granulocyte-colony stimulating (G-CSF) factor was administered in order to reduce neutropenia related to docetaxel. Docetaxel was administered intravenously at a dose of 100 mg/m2, on day 1, and it was repeated every 3 weeks. G-CSF was administered for primary prophylaxis of neutropenia at the standard dose of 30 mg/day from day 4 to day 10 of each cycle. The treatment was tolerated well and 5 (21.7%) partial responses were obtained. The median time to progression and the median survival time were 3 and 5 months, respectively. The main side effect noted was fatigue, the intensity of which was grade 2 in 43.4% of cases and grade 3 in 8.7% of patients, respectively. One patient (4.3%) had grade 4 cutaneous toxicity. No cases of grade 4 neutropenia were reported. In conclusion, docetaxel is active when used for salvage chemotherapy in advanced NSCLC whilst concurrent primary prophylactic administration of granulocyte-colony stimulating factor seems to decrease severe neutropenia. PMID- 10810402 TI - Alteration of cellular mediated cytotoxicity, T cell receptor zeta (TcR zeta) and apoptosis related gene expression in nasopharyngeal carcinoma (NPC) patients: possible clinical relevance. AB - We have investigated apoptosis related gene expression in tumour cells, phenotype and function of blood mononuclear cells at diagnosis in relation to clinical response in three patients with nasopharyngeal carcinoma (NPC). We have focused our study on the Epstein Barr virus latent membrane protein-1 (LMP-1) and Bcl-2 expression in the tumour cells, the essential signal-transducing zeta molecule of T cell receptor (TcR zeta) and cellular mediated cytolysis of the blood mononuclear cells. The carcinoma cells of the patients were Bcl-2 negative. They were heterogeneous with regard to the expression of LMP-1 and the number of proliferating or apoptotic cells. Decrease in the expression of mature T cells (CD3, CD4, and CD8), TcR zeta and cellular mediated cytotoxicity was detected in blood mononuclear cells of the patients. IL-2 up-regulated these phenotypes and the cytolytic capacity of the blood mononuclear cells. The patient with LMP-1 negative carcinoma cells, down-regulated TcR zeta expression and impaired IL-2 mediated cytolysis, had the worst clinical outcome. Another patient with low apoptotic, highly proliferating and LMP-1 positive carcinoma cells had recurrent disease only in the irradiated area. Interestingly, NPC with high apoptotic and few LMP-1 expressing cells was detected in the patient with a normal level of TcR zeta expression and cytolytic functions in blood mononuclear cells at the time of diagnosis. After combination treatment with chemotherapy followed by radiotherapy, this patient is still alive with complete remission and disease free at 36 months. Suppression of the immunological functions may occur in NPC patients. Our study suggests that the immunological functions and apoptosis related gene expression in the carcinoma cells may be used as prognostic factors and help in the decision of therapy of patients with nasopharyngeal cancer. PMID- 10810403 TI - 17 beta-Hydroxysteroid dehydrogenase type 1 and type 2 in ductal carcinoma in situ and intraductal proliferative lesions of the human breast. AB - 17 beta-hydroxysteroid dehydrogenases (17 beta-HSDs) are involved in the interconversion of biologically active and inactive sex steroids and are considered to play important roles in the in situ metabolism of estrogen in various estrogen dependent tissues. 17 beta-HSD type 1 catalyzes primarily the reduction of estrone (E1) to estradiol (E2), whereas 17 beta-HSD type 2 catalyzes primarily the oxidation of E2 to E1. However, the possible biological roles of these estrogen metabolizing isozymes in human breast cancer, especially in carcinogenesis of the human breast, have not been examined in detail. Because of the potential roles of estrogens in the early stages of human breast carcinogenesis, we have examined the immunolocalization of 17 beta-HSD type 1 and type 2 isozymes and estrogen receptor alpha(ER alpha) in both normal human breast tissue and in breast cancers, including ductal carcinoma in situ (DCIS), proliferative disease without atypia (PDWA) or fibrocystic disease and atypical ductal hyperplasia (ADH). We also correlated these findings with clinicopathological findings, Ki67 antigen, progesterone receptor (PR), c-erbB-2, and p53. 17 beta-HSD type 2 immunoreactivity was sporadically detected in non proliferative or Ki67 negative ductal epithelia of normal breast, but rarely in breast carcinoma cells. 17 beta-HSD type 1 immunoreactivity was detected in 12/22 (54.5%) PDWA cases, 8/26 (30.8%) ADH cases, and 25/40 (62.5%) DCIS cases, respectively. 17 beta-HSD type 1 immunoreactivity was not statistically correlated with the age of the patients, Ki67 labeling index (LI), and PR LI, p 53 and c-erbB-2 immunoreactivity. There was no significant correlation between ER alpha LI and 17 beta-HSD type 1 immunoreactivity. There was a positive correlation between ER alpha and Ki67 LI in PDWA, whereas a negative correlation was detected between ER alpha and Ki67 LI in DCIS. There was no correlation between ER alpha and Ki67 LI in ADH. These results suggest that in human breast epithelial cells, development of ADH and DCIS may be associated with the loss and/or deviation of oestrogen dependent regulation of cell proliferation. PMID- 10810404 TI - 89Strontium in the management of painful sceletal metastases. AB - PURPOSE: To make a review of the literature of 89strontium-chloride and a retrospective study of time to palliative intended external irradiation, number of portals and overall-survival after 89strontium-chloride therapy. RESULTS: In total 93 patients were treated 116 times with 89strontium. The patients with prostatic carcinoma received 91% of all 89strontium therapies. Median over-all survival was 10 months after injection. In those cases when 89strontium was given before palliative radiotherapy, the average of total number of local fields was significantly lower (1.1 versus 4.1) compared to those cases where local fields preceded 89strontium therapy. However, time to 89new external irradiation after 89strontium injection was equal between these groups (3.8 versus 2.9 months). CONCLUSION: A review of literature conclude that 89strontium is effective for the reduction of pain originating from osteoblastic metastases. It also reduce the need for external radiotherapy and therefore is cost-effective. However, 89strontium is more effective in an early phase of the metastatic disease and preferably as an adjuvance to external radiotherapy. PMID- 10810405 TI - The role of bone SPET study in diagnosis of single vertebral metastases. AB - The spine is the preferential site of metastases from several neoplasms. In the past years whole body bone scan (BS) with 99mTc-diphosphonates has been considered the first choice in detecting the skeletal involvement. However the presence of vertebral non-neoplastic pathology in oncologic patients can cause several false positive results and this increases the difficulty in defining the etiology of a focal uptake. Nowadays, technological development has provided new gamma cameras, which are able to perform tomographic acquisition (single photon emission tomography, SPET). This technique allows one to better define the anatomical location of the areas of increased uptake. In our study, 81 cancer patients, with suspected single skeletal metastases not defined by BS, were studied by SPET. The skeletal involvement was confirmed during at least 12 months follow up by means of clinical, radiological and nuclear medicine examinations. The overall malignant bone alterations were 14 while the benign ones were 67. The performances of SPET were: diagnostic sensitivity 92.8% (13/14), specificity 92.5% (62/67) positive predictive value 72.2% (13/18), negative predictive value 98.4% (62/63), accuracy 92.6% (75/81). Our conclusion is that bone SPET proved to be a very reliable tool in differentiating benign disease from metastatic involvement. PMID- 10810406 TI - Radiation therapy for T1 glottic cancer: involvement of the anterior commissure. AB - The purpose of this study was to assess the prognostic significance of the anterior commissure (AC) involvement in radiation therapy for T1-stage glottic squamous cell carcinoma. PATIENTS AND METHODS: Sixty-four patients comprising 50 with AC negative tumor and 14 with AC positive tumor, were retrospectively analyzed. Irradiation was administered using parallel opposed lateral portals and wedge filters from 60 to 70 Gy with 2 Gy daily fraction. The tumor doses were estimated by a computer or by an ionization chamber and a thermoluminescence dosimeter. RESULTS: The 5-year local control rates of the AC negative and positive group were 89% and 58%, respectively. The actual dose on the AC was slightly lower than the expected dose which was calculated by a computer. CONCLUSION: The AC involvement was one of poor prognostic factors. The under dosage which was caused by using wedge filters, might have had an effect on the local control. PMID- 10810407 TI - Immunohistochemical staining of human alpha-defensin-1 (HNP-1), in the submandibular glands of patients with oral carcinomas. AB - The purpose of this study was the immunohistochemical localization and distribution of HNP-1 in the submandibular glands of patients with oral carcinomas. Tissue sections were embedded in paraffin, and HNP-1 was immunostained by the streptavidin-biotin coupled peroxidase method. Striated duct cells in the submandibular glands were stained with anti-defensin antibody. Neutrophils and capillary intimal cells were also stained. Defensins (HNPs) are peptides that occur in neutrophils and protect against bacteria and tumor cells. Human alpha-defensin-1 (HNP-1) is such a peptide, possessing both antimicrobial and cytotoxic activities. The presence of HNP-1 in striated duct cells in the submandibular glands of oral cancer patients, suggests a likely role in tumor immunity, for this peptide. PMID- 10810408 TI - Missense alterations of BRCA1 gene detected in diverse cancer patients. AB - The mutations in the breast cancer susceptible gene BRCA1 are responsible for about 50% of inherited breast cancers and confer increased risk of breast and ovarian cancer to its carriers. BRCA1 gene mutations may also be related with other types of cancers such as prostate cancer and colorectal cancer. The goal of this study was to investigate if BRCA1 mutation could be detected in diverse types of cancers. We used PCR-NIRCA and PCR-SSCP methods for screening the BRCA1 mutation hot regions, exons 2, 5, 11, 16 and 20. The positive samples were sequenced to confirm the nature of the mutations. We have identified a rare sequence variant, A3537G (Ser 1140Gly) in a B cell lymphoma patient and two polymorphisms, A1186G (Gln356Arg) in a brain cancer patient and A3667G (Lys1183Arg) in a germline tumor patient. In conclusion, 3 missense alterations of BRCA1 gene have been identified in cancers other than breast cancer. PMID- 10810409 TI - First report of lymphatic mapping with isosulfan blue dye and sentinel node biopsy in cervical cancer. AB - BACKGROUND: Sentinel lymph node status provides important information about the status of the regional nodes in various malignant tumors. Our report describes a method of identifying the sentinel lymph nodes in cervical cancer. PATIENTS AND METHODS: In three cases of early cervical cancer, isosulfan blue dye was injected paracervically into each lateral fornix immediately before surgery. RESULTS: In all cases we identified two to three blue stained (sentinel) lymph nodes located either at the iliac artery or in the obturatory space. The blue colored nodes were positive for disease, all other pelvic lymph nodes removed were negative. CONCLUSIONS: Our findings demonstrate that preoperative lymphatic mapping with vital blue dye is an easy to perform technique to visualize sentinel lymph nodes in cervical cancer. Sentinel lymph node status may be representative of the pelvic lymph node status in cervical cancer and thus could provide important information for further treatment. PMID- 10810410 TI - Changes in cell proliferation and apoptosis during local progression of prostate cancer. AB - To determine the changes in biological features of the prostate during the course of local recurrence of prostate cancer after endocrine therapy, histologic grade, proliferating activity and apoptotic indices were examined in prostate specimens obtained before treatment and at recurrence. A total of 16 patients, who had received endocrine therapy and eventually recurred in the prostate, were evaluated. Histologic grade was determined by the method of Gleason and the number of proliferating cells and apoptotic cells were counted. Tumors with a high grade Gleason score remained at a high grade. A statistically significant increase in the number of Ki-67 positive cells was observed from pretreatment biopsy to local recurrence. On the other hand, the apoptotic index decreased during progression. Patients with a higher number of Ki-67 positive cells before the initial treatment had a poorer prognosis than those with a lower number of Ki 67 positive cells. In conclusion, prostate cancer shows an increase of malignant potential as assessed by the number of Ki-67 positive cells, whilst the decrease in apoptosis might play some role in the course of progression. PMID- 10810411 TI - Serrated adenoma: a clinicopathological, DNA ploidy, and immunohistochemical study. AB - AIMS: Serrated adenoma (SA) is a relatively newly defined entity of colorectal neoplasm. In this study, we examined the cell proliferation, DNA ploidy, and clinicopathological features of SA in order to investigate its biological features. METHODS AND RESULTS: We reviewed 10,532 polypectomy specimens of the colorectum obtained from Japanese cases between 1974 and 1998 at Tohoku University Hospital. In total, 193 cases of SA were detected. We first examined clinical features of these cases by reviewing the charts, and then studied cell proliferation using immunohistochemistry of Ki-67 and topoisomeraseIIa, p53 immunoreactivity and DNA ploidy. Results were subsequently compared with those of tubular adenoma (TA) and hyperplastic polyp (HP). Mean size of SA (8.6 +/- 4.6 mm) was significantly larger than those of TA (7.3 +/- 4.6 mm) and HP (5.6 +/- 3.0 mm). More than 80% of SA were protuberant in macroscopic appearance. SA was located predominantly in the sigmoid colon and rectum. Incidences of concomitant carcinoma in HP, SA and TA were 0.4% (1 out of 263), 4.1% (8 out of 193) and 10.3% (809 out of 7838), respectively. Labeling indices for Ki-67 and topoisomeraseIIa in HP, SA and TA were as follows: Ki-67--24.2%, 30.8%, 39.5% and topoisomeraseIIa--15.3%, 16.1%, 23.9%, respectively. In SA, p53 immunoreactivity was detected in the intramucosal carcinoma co-existing with the serrated component. Two out of the ten SA cases examined demonstrated non-diploid patterns of DNA ploidy. CONCLUSION: SA is a distinct colorectal neoplastic lesion with the potential of malignant transformation similar to that of tubular adenoma. PMID- 10810412 TI - Detection of occult tumor cells in peripheral blood from patients with small cell lung cancer by reverse transcriptase-polymerase chain reaction. AB - The reverse transcriptase-polymerase chain reaction (RT-PCR) of tumor-specific or -associated genes is a sensitive assay for detecting a minimal number of tumor cells in peripheral blood (PB) or bone marrow (BM). In this study, we determined whether mRNA of bombesin receptors is detectable in PB or peripheral blood progenitor cell (PBPC) samples from patients with small cell lung cancer. Among three bombesin-like peptide receptors, we used the neuromedin B receptor (NMB-R) gene as a target, because of the most frequent expression on SCLC cell lines. The lower limit of detection was one tumor cell in one million normal PB cells and there was no detection in normal PB or BM cells unlike a cytokeratin 19 gene. The NMB-R gene was detected in 14 (31.8%) of 44 PB samples from patients with SCLC at diagnosis and 2 (15.4%) of 13 samples of PBPC collected during a recovery phase after chemotherapy followed by administration of G-CSF (filgrastim). At diagnosis, patients whose PB was positive for the NMB-R gene had a significantly shorter survival than those who were negative. Our observation suggests that this assay may be useful in diagnosing metastatic disease and monitoring minimal residual disease in patients with SCLC. PMID- 10810413 TI - Tumor size does not have prognostic significance in stage Ia NSCLC. AB - PURPOSE: We aimed to determine whether the prognosis of non-small cell lung cancer (NSCLC), with lesions 2 cm or less in diameter, was better than that of Stage Ia lung cancer having a diameter of more than 2 cm. METHODS: We reviewed 64 cases of NSCLC which were preoperatively diagnosed as TINOMO. The tumors were classified into 2 groups on the basis of a tumor diameter set at 2 cm. After comparison of background factors, we compared the prognoses of the groups and determined the prognostic factors in Stage IA NSCLC. RESULTS: No differences were observed in the survival rate between the 2 groups. Statistically significant factors for the survival period in Stage IA NSCLC were serum carcino-embryonic antigen (CEA), residual tumor condition and lymph node metastasis. Those cases where CEA was abnormal were all in Stage II or inllmore advanced stage. CONCLUSION: It is not justifiable to sub-classify Stage IA NSCLC on the basis of a tumor diameter of 2 cm. The present study revealed that the most important preoperative prognostic factor was serum CEA. PMID- 10810414 TI - Treatment of recurrent Kaposi's sarcoma of an AIDS patient with weekly paclitaxel. AB - Paclitaxel was recently recognized as an active chemotherapeutic agent for acquired immunodeficiency syndrome (AIDS)-associated Kaposi's sarcoma (KS). However, the best way to administer paclitaxel in AIDS-KS patients remains unknown. Herein, we reported an AIDS-associated KS patient whose disease progressed on the first-line chemotherapy with doxorubicin and bleomycin, but later responded well to weekly 1-hour infusion of 70 mg/m2 paclitaxel. It is particular noteworthy that this weekly dosing schedule resulted in almost negligible toxicities. The authors suggested a prospective study of weekly paclitaxel for AIDS-KS should be started as soon as possible. PMID- 10810415 TI - Carcinoma of the extrahepatic biliary system: correlation of clinical, pathological, histological and DNA-cytometric parameters with prognosis. AB - BACKGROUND: Studies concerning the prognosis of all carcinomas of the extrahepatic biliary system together (EBC) do not exist, so far. A detailed and valid histopathological grading-system is lacking and the prognostic relevance of DNA-cytometric parameters is not clear. METHODS: 81 patients with EBC underwent surgical treatment. Clinical, pathological, histomorphological and DNA-cytometric parameters were evaluated. The influence on prognosis was tested in univariate and multivariate analysis. RESULTS: Apart from significant perioperative mortality (17%) survival was markedly dependent on the presence of tumor remnants in resection margins. After incomplete and complete resection, the median survival was 5 and 19 months, respectively (p < 0.0001). Further parameters with significant prognostic relevance were tumor localization, pTNM-stage, period of complaints, presence of gallstones, histological type and DNA-index. Multivariate analysis however demonstrated only two independent prognostic factors: status of resection margins and tumor localization. CONCLUSIONS: Histomorphological- and DNA-cytometric parameters failed to demonstrate an additional significant influence on the prognosis in EBCs. Apart from perioperative mortality, the survival of patients with EBC predominantly depends on tumor localization and the status of resection margins. PMID- 10810416 TI - Usefulness of urinary nuclear matrix protein 22 (NMP22) as a marker for transitional cell carcinoma of the bladder. AB - OBJECTIVES: The purpose of this study was to evaluate an immunoassay for urinary nuclear matrix protein, NMP22, as a novel marker for urothelial cancer. PATIENTS AND METHODS: NMP22 values were determined for 71 patients and 21 healthy volunteers. Each subject provided a single (3 voids) urine sample for analysis at the time of entry into the study. Each sample was assayed for levels of NMP22. RESULTS: When the cut-off value was set at 10 U/ml, the positive rate for urinary NMP22 in bladder cancer was 37.8% (17 out of 45), whereas that in post-treatment cases and benign diseases was 30.8% (8 out of 26) compared to 14.3% (3 out of 21) for healthy volunteers. This cut-off value provided a sensitivity of 37.8% and a specificity of 80.9%. In the bladder cancer group, NMP22 levels were related to tumor size, shape, grade and stage. CONCLUSIONS: Despite the many reports that suggest NMP22 as a promising urinary marker for monitoring transitional cell carcinoma, this study does not support its usefulness as a substitute tool for urinary cytology in the control of bladder tumors. PMID- 10810417 TI - The clinical value of p21WAF1/CIP1 expression in superficial bladder cancer. AB - The clinical value of p21WAF1/CIP1 in superficial bladder cancer remains controversial. To address the question, we examined the expression patterns of p21 and p53 gene products and compared for their significance in a total of 89 cases of superficial (pTa/pT1) bladder cancer. Over-expression of p21 was detected in 32 of 89 (36%) tumors. But, the expression status did not correlate with biological indicators or clinical outcome (p > 0.1, respectively). Factors predicting clinical outcome were multiplicity for tumor recurrence (p = 0.0002) or patient survival (p = 0.03), and the histological grading for disease progression (p = 0.02) or patient survival (p = 0.05). Taking into account the p53 status, a trend approaching better prognosis for p53+p21+ tumors was observed compared with that of p53+p21- bladder cancer (p = 0.08). Our data indicate that evaluation of p21 status does not provide better prognostic information compared with conventional biological indicators of superficial bladder cancer. Maintenance of p21 appears to abrogate the deleterious effects of p53 alterations in the tumorigenesis of human bladder. PMID- 10810418 TI - Phase II study of continuous 120-hour-infusion of mitomycin C as salvage chemotherapy in patients with progressive or rapidly recurrent gastrointestinal adenocarcinoma. AB - The aim of the study was to evaluate the therapeutic activity and safety of continuously infused (c.i.) mitomycin C in patients with metastatic gastrointestinal adenocarcinomas recurring after or progressing during chemotherapy. The patients were treated with mitomycin C 20 mg/m2 intravenously over a time period of 120 hours, followed by a 3-week rest. Seventy-eight patients were entered into the study. All the patients were assessable for toxicity and 72 for response evaluation, having completed at least one full course of chemotherapy. Among 52 colorectal cancer patients one single objective response (2%) was observed. Disease stabilisation was achieved in 11 patients (23%). Median survival time was 4.7 months (CI95%, 1.9 to 8.6). In 20 evaluable patients with gastric cancer, one complete and 5 partial remissions were observed [ORR: 30% (CI95%, 9.1-50.9%)]. The median response duration was 2.1 months (range, 2 to 6), and the median survival time 3.6 months (CI95%, 2.1 to 6.0). WHO degree III/IV mucositis, diarrhea and fever/infection each occurred in 6% of the patients. Thrombo- and leukocytopenia (WHO degree III/IV) occurred in 19% and 6% of the patients, respectively. Mitomycin C has some single-agent activity in advanced gastric cancer but no substantial efficacy was seen as in heavily pretreated colorectal carcinoma. PMID- 10810419 TI - Human papilloma viruses in oral lesions. AB - Oral mucosa biopsies from 53 patients with different oral diseases, and from 12 healthy control patients, were examined by the PCR-technique using the L1 consensus primers and type-specific primers for HPV 6/11, 16 and 18. Three out of 24 (12.5%) oral cancer biopsies were HPV positive, 1 for HPV 18, 1 for HPV 16 and 6/11 and 1 for none of the specific primers. Six out of 22 (27.3%) lichen planus were HPV positive, 5 for HPV 18 and 1 for none of the specific primers. Seven leukoplakias were included in the study and 2 (29.6%) were positive for both HPV 16 and 6/11. None of the 12 control patients was HPV positive. No statistical difference between the use of tobacco and alcohol and HPV prevalence was found. An association between HPV infection and oral lesions was demonstrated in the studied population but the pathogenic influence of HPV infection remains unclear. PMID- 10810420 TI - Serum bFGF (basic fibroblast growth factor) and CA 15.3 in the monitoring of breast cancer patients. AB - BACKGROUND: Basic fibroblast growth factor (bFGF) is a potent angiogenetic factor which may influence breast cancer evolution. MATERIALS AND METHODS: Serum bFGF, (cut-off 10 pg/ml), was assayed in 166 breast cancer patients at all stages and compared with CA 15.3. RESULTS: In 99 pre-treatment (PT) sera, 39/99 (39.4%) were bFGF positive, 9/99 (9.1%) CA 15.3 positive (> 30 U/ml), and not correlated. No correlations were found between bFGF and age, menopausal status, TNM or pTNM, histology, SBR grading or steroid receptors. A postoperative decline in bFGF positivity, from 30.8 to 7.7% (n = 39), was observed. An abnormal CA 15.3 after primary treatment (n = 2/39) was of bad prognosis (P < 0.0001), whereas positive bFGF (n = 3/39) had no univariate prognostic value (median follow-up 5.5 years). During follow-up, positive bFGF was recorded in 6/92 (6.5%) disease-free patients (DFS), 13/15 (86.7%) regressions, 8/16 (50.0%) stable disease, and 46/67 (68.7%) progressive disease (significant differences between PT or DFS and post recurrence levels (P < 0.001), and between relapse before and after treatment (P = 0.002)). CONCLUSION: Serum bFGF is more often elevated before treatment or after relapse than in DFS, and rises under systemic treatments. Its pattern of variations does not add to CA 15.3 for breast cancer monitoring. PMID- 10810421 TI - Diagnosis and survey of abnormal/atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesions: a retrospective study. AB - The new Bethesda System (BS) terminology has opened a series of problems about the Abnormal/Atypical Squamous Cells of Undetermined Significance (ASCUS) and Low Grade Squamous Intraepithelial Lesion (L-SIL) categories, particularly on their treatment and follow-up. Moreover in these field a non negligible portion of lesions progress to High-Grade Lesion (H-SIL). With the aim of comparing the data, we examined samples observed in our Ambulatories with 6-12 months follow up. We observed retrospectively 11.197 Pap test from January 1995 to June 1997, mostly first visits. All received a Pap test, colposcopy and gynaecological examination. Biopsy and histological standard examinations were performed when necessary. Alterations classified as mild epithelial abnormalities, of both ASCUS and L-SIL categories, were observed in 146 cases: 78 ASCUS and 68 L-SIL. Of these 48 were CIN 1/mild dysplasia (25 HPV associated) and 20 were HPV lesions, according to BS. All the cases with persistent ASCUS and L-SIL underwent a second control. In these 45 cases we observed 2 new cases of H-SIL and one of L-SIL; so 82.3% and 30.7% of ASCUS and L-SIL regressed to negatives. In the stable group 11.8% and 60.4% were ASCUS and L-SIL. Moreover 5.9% of ASCUS and 4.8% of L-SIL progressed to H-SIL. No cases of invasive cancer were observed. Our data show that ASCUS and L-SIL diagnosis can identify 6% and 4.8% of H-SIL respectively. In addition 17-18% of ASCUS were stable or progressed. These two categories, as already demonstrated in other studies, are mostly at risk. PMID- 10810422 TI - Elevated serum CA-125 levels in patients with nephrotic syndrome-induced ascites. AB - CA-125 is a sensitive, but not a specific, tumour marker especially used in the diagnosis and follow-up of ovarian cancer. Because of the elevated levels of CA 125 encountered during the etiological investigation of nephrotic syndrome (NS) a clinical study was designed to investigate the probable relationship between elevated CA-125 levels in patients with NS, with and without ascites. Twenty-four patients with NS due to non-tumoural pathologies, aged 24 to 56 (15 females and 9 males), were investigated. We detected elevated levels of serum CA-125 (275.92 +/ 154.71 U/mL) in 14 (70%) NS patients with ascites (p < 0.05). In the other 10 NS patients without ascites (Group 2) and in the control group, consisting of 52 age and sex-matched healthy volunteers, the serum CA-125 levels were found to be 13.60 +/- 5.12 U/mL and 8.50 +/- 5.02 U/mL, respectively (p > 0.05). There was no statistically significant difference between the control group and the patients without serosal fluids (p > 0.05). We concluded that serum CA-125 levels were elevated in NS patients with ultrasonographically detected ascites in the absence of an ovarian tumour or other diseases known to increase the levels of CA-125. PMID- 10810423 TI - Immunohistochemical study of basaloid squamous cell carcinoma, adenoid cystic and mucoepidermoid carcinoma in the upper aerodigestive tract. AB - Non-squamous cell carcinoma is a rare but distinct neoplasm of the upper aerodigestive tract. Among these carcinomas, basaloid-squamous cell carcinoma (BSCC) has frequently been confused with adenoid cystic carcinoma (ACC) and mucoepidermoid carcinoma of the upper aerodigestive tract. In this study, we examined immunohistochemically the expression of differentiation-related substances, including cytokeratin (CK) subtypes, p53 and p27, and cell adhesion related molecules E-cadherin and alpha-catenin to clarify the biological features of these neoplasms. We studied seven cases of BSCC of the oesophagus, five cases of ACC and seven cases of mucoepidermoid carcinoma. Squamous cell carcinoma and adenocarcinoma of the oesophagus and trachea were also studied for comparison. Among the cytokeratin subtypes examined, CK14, CK17 and CK19 immunoreactivity was detected in BSCC. ACC and mucoepidermoid carcinoma were immunopositive for CK8, CK14 and CK17 and for CK8, CK14, CK17 and CK19, respectively. These findings suggest that CK subtypes, especially CK8, CK14 and CK17, are useful in differentiating these malignancies. BSCC was more frequently associated with decreased E-cadherin and alpha-catenin immunoreactivity than ACC and mucoepidermoid carcinoma. Nuclear p53 immunoreactivity was detected more frequently in BSCC (5 out of 7) than in ACC (2 out of 5) and mucoepidermoid carcinoma (4 out of 7). There were no significant differences in p27 immunoreactivity among these carcinomas. Carcinoembryonic antigen (CEA) immunoreactivity was detected in mucoepidermoid carcinoma (2 out of 7), SCC (8 out of 11) and adenocarcinoma (9 out of 9), but it was not detected in BSCC (7) or ACC (5). Carbohydrate antigen 19-9 (CA19-9) immunoreactivity was detected only in mucoepidermoid carcinoma (4 out of 7) and adenocarcinoma, but not in BSCC, ACC, or SCC. These findings indicate that BSCC, ACC and mucoepidermoid carcinoma are distinct neoplasms arising in the upper aerodigestive tract. In addition, decreased expression of E-cadherin and alpha-catenin proteins and increased p53 expression in BSCC may be correlated with aggressive behaviour. PMID- 10810424 TI - Bcl-2 immunostaining: a way to finding unresponsive postmenopausal N+ breast cancer patients. AB - BACKGROUND: Bcl-2 staining positivity has shown limited prognostic value. However, we have decided to study this issue in the era of mammography screening, and adjuvant treatment protocols. METHODS: Paraffin sections of 414 breast cancers were stained for bcl-2 and staining intensity graded. Association of bcl 2 with established prognosticators was analysed with chi 2 test and odds ratios in 2 x 2 tables. Kaplan-Mayer analysis and Cox's regression were used to evaluate the prognostic value of bcl-2 and other prognosticators. RESULTS: Bcl-2 immunostaining was associated with tumor size, lymph node status, histological type, multivariate prognostic index, standardized mitotic index, Ki-67 fraction, DNA-index, proportion of cells with DNA above 5c, estrogen receptor status, and histological grade. ER status showed the best association with bcl-2 positivity (odds ratio 11.3, 95% CI 5.6-22.7). In the whole group of patients bcl-2 positivity was not an independent prognosticator. However, among N+ patients bcl 2 staining was significant, and among postmenopausal N+ patients bcl-2 immunostaining was a stronger independent prognosticator than tumor size. CONCLUSIONS: The prognosis of N+ breast cancers can potentially be evaluated with bcl-2 positivity, in association with tumor size, and mitotic activity. Among postmenopausal N+ patients, most of whom have received anti-estrogen therapy, bcl 2 positivity is an independent prognosticator. Also, the close association of bcl 2 positivity with ER status supports the view that bcl-2 negativity reveals a patient group which might benefit from additional treatment in association with anti-estrogen therapy. PMID- 10810426 TI - Estimation of glutathione S-transferase and its Pi isoenzyme in tumor tissues and sera of patients with ovarian cancer. AB - OBJECTIVES: The Pi class GST is the most ubiquitous of the human GST family, being expressed in different tissues. However, its role in ovarian cancer is still poorly defined. To establish normal and tumor related changes, the levels of total GST as well as its isoenzyme GST Pi, were therefore measured in ovarian tissue samples and the corresponding serum specimens obtained from patients undergoing primary surgical treatment for their disease (n = 68). The total GST activity was spectrophotometrically determined utilizing 1-Chloro- 2,4 dinitrobenzene (CDNB) as the substrate. The GST Pi isoenzyme was measured by ELISA and immunohistochemical methods. RESULTS: The total GST activity, the level of GST Pi isoenzyme, the DNA content and S phase fraction were significantly higher in the malignant than in non-malignant ovarian tissues. In the malignant group the values of both cytosolic total GST and GST Pi were significantly correlated (r = 0.56, P = < 0.01). In spite of the absence of a significant correlation with a number of important prognostic features including the patient age, FlGO stage, DNA content and S-phase fraction, both total GST and GST Pi were significantly higher in the grade I than in grades II and III malignant tumors. Additionally, the total GST was significantly lower in the serous than non-serous malignant tumors. CONCLUSION: In the malignant group, although the elevation of blood content of GST and its Pi isoenzyme was not as significant as that of the tissue content, the fact that higher serum values of patients with some cancers often reverted to the normal range after treatment of the cancer suggested the direct derivation of these enzymes from tumor tissues. Thus, follow up of elevated serum GST and GST Pi levels may be useful for monitoring ovarian cancer patients during the course of treatment. PMID- 10810425 TI - Impact of bacterial eradication on the cell proliferation and p53 protein accumulation in Helicobacter pylori-associated gastritis. AB - BACKGROUND: This study investigates the cell proliferation and the expression of p53 protein in Helicobacter pylori (H. pylori)-associated gastritis and assesses the effect of bacterial eradication on these epithelial factors. MATERIAL AND METHODS: Seventy-nine patients with H. pylori-associated gastritis were randomized into the control group (n = 38) and anti-H. pylori group (n = 41). Each patient received endoscopic examinations with gastric biopsy before and 8 weeks after the treatment. The specimens from gastric antrum were immunostained for monoclonal antibodies against the proliferating cell nuclear antigen (PCNA) and p53 protein. RESULTS: In the control group, the total labeling index (L.I.) of PCNA and the positive index (P.I.) of p53 in the whole foveolar epithelium were unchanged after treatment. In the anti-H. pylori group, 35 of 41 cases (85.3%) achieved eradication of H. pylori. Amongst the H. pylori-eradicated cases, the total L.I. of PCNA in the whole foveolar epithelium did not meaningfully alter after H. pylori elimination (p > 0.05). However, a significant reduction of L.I. was observed in the middle compartments of the gastric pits (before vs. after treatment: 14.0 vs. 7.3, p < 0.05). With regard to the p53 expression, the P.I.s were significantly decreased in the whole foveolar epithelium (before vs. after treatment: 0.57 vs. 0.17, p < 0.05) and in each compartment of the gastric pits (before vs. after treatment: [upper compartment]: 0.34 vs. 0.15, p < 0.05; [middle compartment]: 0.67 vs. 0.23, p < 0.05; [lower compartment]: 0.71 vs. 0.20, p < 0.05) after eradication of H. pylori. CONCLUSIONS: Bacterial eradication reverses the hyperproliferating status of the foveolar epithelium in patients with H. pylori gastritis and leads to a decrease in p53 accumulation in the epithelial cells. PMID- 10810427 TI - Anticancer chemosensitivity profiles of human breast cancer cells assessed by in vitro DNA synthesis inhibition assay. AB - The present study was designed to assess the profile of the chemosensitivity of breast cancer cells and to screen effective agents for combination regimens. Chemosensitivity to anticancer agents was assessed by the 3H-thymidine incorporation assay, as the rate of inhibition of DNA synthesis in 145 samples (88 primary and 57 metastatic or recurrent lesions) from 136 patients with breast cancer. The correlations of the anticancer agents with various clinicopathological factors were analysed. The effectiveness of the agents was classified as a rate of inhibition on log scale as follows: highly sensitive (> or = 30%), moderately sensitive (25-30%), slightly sensitive (20-25%), resistant (< 20%). The chemosensitivity of breast cancer showed variations according to tumor location: primary lesions seemed to be slightly sensitive to carboquone (CQ), adriamycin (ADR), and cytosine arabinoside (Ara-C); nodal involvement was moderately sensitive to CQ and slightly sensitive to Ara-C, 5-FU, ADR, mitomycin C (MMC), and cisplatin (CDDP); malignant effusions were highly sensitive to ADR, moderately sensitive to CQ, and slightly sensitive to Ara-C and CDDP; local recurrences were slightly sensitive to Ara-C, CQ and 5-FU; vincristine (VCR) and nimustine chloride (ACNU), however, seemed to be ineffective against breast cancer. There were significant correlations in chemosensitivity between most agents, but no correlation was found between 5-FU and CDDP, 5-FU and ACNU, MMC and VCR, ADR and CDDP, ADR and VCR, and ADR and ACNU. There were no differences in chemosensitivity between stages of primary lesions or between estrogen receptor-positive and -negative tumors. In 10 patients, simultaneous nodal involvement was more sensitive to the agents than were primary lesions, and the correlation of chemosensitivity to ADR and CQ between such lesions was significant. On the other hand, there was no significant difference or correlation of chemosensitivity between the original lesions and recurrent ones after chemotherapy. The heterogeneity and homogeneity in the chemosensitivity of breast cancer suggested not only the necessity of patient-specific chemotherapy based on a sensitivity assay, but also the usefulness of choosing agents for widely-applicable combination regimens against breast cancer. PMID- 10810428 TI - Administration of tamoxifen in the perioperative period to patients with breast carcinoma prolongs axillary fluid drainage. AB - Compelling evidence points to an estrogen receptor independent mechanism of action of tamoxifen in the extracellular matrix, in addition to its action via the estrogen receptors. We retrospectively studied 380 patients who underwent curative resection for primary breast cancer in our institution from January 1994 to December 1998, of which 227 received tamoxifen in the perioperative period and the remaining 153 never received tamoxifen or delayed the initiation of treatment for at least two weeks following the operation. The administration of tamoxifen in the perioperative period resulted in the prolongation of axillary drainage (mean 10.07 SD 4.18 days vs mean 8.33 SD 2.85 days), which was statistically significant for patients younger than 70 years. There was no difference in the duration of fluid drainage in relation to the number of positive nodes, except in cases of more than 9 nodes involved by the tumor. We assumed that tamoxifen causes a delay in would healing through the secretion of active transforming growth factor beta(TGF-beta), which is the principle negative growth modulator and which can be secreted from epithelial and stromal elements, independently of hormonal receptor status. PMID- 10810429 TI - Phase I clinical trial of liposomal daunorubicin in hepatocellular carcinoma complicating liver cirrhosis. AB - Chemotherapy has been proposed for patients with hepatocellular carcinoma (HCC) associated with well-compensated cirrhosis who are unsuitable for locoregional treatments. Anthracyclines are the most active agents against HCC, although their toxicity is often unpredictable; thus, there is a need for new active drugs with a safe toxicity profile. The liposomal formulation of the anthracycline daunorubicin has low systemic toxicity and is taken up strongly by the liver. We started a phase I study with liposomal daunorubicin (starting dose 80 mg/m2 every 21 days) in patients with hepatocellular carcinoma and Child-Pugh stage A or B liver cirrhosis. At the starting dose, we recorded dose-limiting toxicities (1 hyperbilirubinemia, 1 prolonged prothrombin time, 1 persisting grade 3 neutropenia) in 3 out of 4 patients. Thus, according to protocol, we went down to the dose level of 60 mg/m2 but still 2 out of 3 patients had unacceptable toxicity (1 hypertransaminasemia, 1 hyperbilirubinemia with encephalopathia). Finally, we treated 4 more patients at the dose level of 40 mg/m2 and again we recorded liver toxicity in three of them. Overall haematological toxicity was mild and significant non-haematologic toxicities, other than hepatic, were not recorded. The toxicity profile observed warns against further assessment of liposomal daunorubicin in patients with hepatocellular carcinoma and liver cirrhosis. PMID- 10810430 TI - Chronomodulated infusion of cisplatin, 5-fluorouracil and folinic acid: lack of activity in advanced colorectal cancer. AB - BACKGROUND: The chronomodulated infusion of 5-FU, FA and oxaliplatin allows a significant increase in dose intensity and antitumor efficacy in patients with metastatic colorectal cancer. Here we investigated if substitution of oxaliplatin with cisplatin produced a similar antitumor activity in previously untreated patients with advanced colorectal cancer. METHODS: We enrolled 21 consecutively evaluated ambulatory patients with metastatic colorectal cancer. Each treatment cycle consisted of a 5-day course of continuous chronomodulated venous infusion of drugs. Daily doses were 600 mg/m2 5-FU, 150 mg/m2 FA (L-form), and 12 mg/m2 cisplatin. The cycles were repeated every 21 days. RESULTS: All patients completed at least 3 cycles. Overall a total number of 105 cycles were administered. One partial response (lasting 3 months) and 13 stable disease (lasting from 3 to 12 months) were observed. The remaining 7 patients had progression of the disease. Hematologic and gastrointestinal toxicity was always < or = G2 in all cycles. CONCLUSIONS: The results of this study discourage the substitution of cisplatin for the more active compound, oxaliplatin, in a chronomodulated schedule of infusion with 5-FU and FA in patients with metastatic colorectal cancer. PMID- 10810431 TI - Relationship between loss of heterozygosity at microsatellite loci and computerized nuclear morphometry in hepatocellular carcinoma. AB - Loss of heterozygosity (LOH) at 4 microsatellite loci (chromosomes 8p23, 8q24, 13q14 and 17p13) and the nuclear morphometric data were analyzed in 28 tumors from 28 patients with hepatocellular carcinoma (HCC). LOH was detected in 63.2% of the tumors with marker D8S264 (chromosome 8p23), in 52.6% with marker D8S1801 (chromosome 8q24), in 25.0% with marker D13S328 (chromosome 13q14), and in 52.4% with marker D17S796 (chromosome 17p13). LOH on 17p13 was most frequent in tumors that had reached more advanced stages (p < 0.05). The frequencies of LOH on 17p13 were significantly higher in tumors with portal invasion of cancer cells or intrahepatic metastasis than those without (p < 0.05). Moreover, capsule infiltration of cancer cells or hepatitis B viral infection were more frequently detected in patients with LOH on 17p13 in their tumors. The mean nuclear area (MNA) and the standard deviation (SDNA) of tumors with LOH on 17p13 were significantly larger than those of the other sites (p < 0.01). No significant relationships were found between the LOH on 8p23, 8q24 or 13q14 of HCC and the clinicopathologic data, MNA or SDNA. Therefore, LOH on 17p13 of HCC correlates with the stage, portal invasion of cancer cells, intrahepatic metastasis, and nuclear morphometry. These findings suggest that the LOH on 17p13 is closely connected to the progression of HCC and that nuclear morphometry is useful as an indicator of genetic alterations in HCC. PMID- 10810432 TI - Elevation of serum hepatocyte growth factor concentration in patients with gastric cancer is mediated by production from tumor tissue. AB - Hepatocyte growth factor (HGF) is a stromal cell-derived cytokine that can stimulate invasion and metastasis of carcinoma cells. Recent studies have shown that the serum HGF concentration is elevated in patients with gastric cancer and may be a useful disease marker. However, the origin of the elevated serum HGF remains undetermined. We investigated the site of HGF production by analyzing the relationships between the HGF expression in tumor tissues, the serum HGF concentrations and inflammation in patients with gastric cancer. The serum and tissue HGF concentrations were measured by an enzyme-linked immunosorbent assay. The serum HGF concentration was higher than the normal cut-off level (0.57 ng/ml) in 44% of the patients. Surgical removal of the tumor significantly reduced the serum HGF concentration, suggesting that the tumor tissue was responsible for the increase. Western blotting analysis showed that the HGF protein was expressed in 20 out of 22 tumor tissues. The concentration of HGF in the tumor tissue was significantly higher than that in normal gastric mucosa. Significant correlation was found between tissue HGF concentrations and serum HGF concentrations. No significant correlation was found between the serum HGF concentration and white blood cell count or C-reactive protein concentration, indicating that the increase in serum HGF is not due to inflammation related to the tumor. These results suggest that the elevated serum HGF concentration in patients with gastric cancer is mediated by production from the tumor tissue. PMID- 10810433 TI - Anti-apoptotic phenotype is associated with decreased locoregional recurrence rate in breast cancer. AB - PURPOSE: Tumor stage and nodal status are the most important factors predicting locoregional recurrence in breast cancer. We wanted to investigate the prognostic value of some newer molecular genetic markers for the occurrence of a locoregional recurrence, in order to improve the selection of patients for locoregional adjuvant therapy. METHODS: Bcl-2, p53, MIB-1, pS2 and CD44v6 were determined immunohistochemically on formalin-fixed and paraffin embedded tumour tissues of 163 patients treated by modified radical mastectomy between 1982 and 1987. Postoperative irradiation was given to 35 patients to the intermammary chain only and to only 13 (8%) patients to the chest wall with or without the regional lymph nodes. Node-positive patients were treated with CAF adjuvant chemotherapy and were randomized for whether or no additional Medroxyprogesteroneacetate (MPA). A multivariate analysis was performed on a number of potential prognostic factors. The risk for locoregional recurrence was estimated using the competing risk approach. RESULTS: After a median period of 7.5 years 28 patients developed a locoregional recurrence. The cumulative incidence of loco-regional recurrence at 10 years was 17%. Bcl-2 and p53 were found to be independent factors predicting locoregional recurrence, whereas a trend was found for MIB-1. Increased Bcl-2 as well as p53 expression were associated with a decreased risk, whereas the increased presence of MIB-1 was associated with an increased risk. CONCLUSION: Results indicate that molecular markers of apoptosis as well as proliferation provide additional information for the risk of locoregional recurrence after modified radical mastectomy. If confirmed, these markers may play a role in the selection of appropriate locoregional adjuvant treatment after primary surgery. PMID- 10810434 TI - Preoperative stereotactic hookwire localization of nonpalpable breast lesions with and without the use of a further stereotactic check film. AB - A series of 394 women underwent 399 stereotactically guided preoperative hook wire localization of non-palpable breast lesions. 250 lesions were localized without checking the position of the needle tip with a further stereo film. Later the position of the localization needle was checked by means of a further stereo film in 149 lesions. If the needle tip was beside the lesion (failed x- or y coordinates), we took a new stereo film. Errors in needle depth (z-coordinate) produced particularly subtle signs. Therefore a polystyrene phantom and a metallic object was used to ascertain how to recognize needle depth error on the stereotactic film. In 51 lesions readjustment of the needle depth was necessary. Measured by our criteria, successful and acceptable localization procedures with or without a check stereo-image film showed no significant differences. Unsuccessful localizations without the check film were significantly higher (16.0%) than with the check film, without readjusting the needle depth (7.1%) or after readjusting the needle depth (9.8%). These unsuccessful localizations were mostly architectural distortions or microcalcifications. Our results suggest that success of the localization procedure is better by using the stereo check film as without it, even in lesions which are difficult to resolve and target on films. However, readjustment of the needle depth did not always lead to successful localization. PMID- 10810435 TI - Knowledge of tumor markers and the psychological consequences of tumor marker sampling in patients with gynecologic cancers. AB - BACKGROUND: The aim of this study was to investigate the extent of knowledge about serum tumor markers in patients suffering from gynecologic cancer. MATERIALS AND METHODS: 360 women with a median age of 60 years (range: 26-88 years) visiting the oncological outpatient clinic of the Department of Gynecology and Obstetrics of the University of Vienna, between February and July 1998, were asked to complete a self-report questionnaire. RESULTS: The majority of patients (85.2%) believed it was important to know about tumor markers and felt safe when they knew the recent level of the tumor marker (71.6%). On the other hand, many patients felt they were insufficiently informed (43%). 88.9% of the patients did not know the recent serum level of the tumor marker. The patients who had been informed by a physician were significantly better informed about tumor markers than women relying on other sources such as nurses, relatives or other patients (p < 0.001). Patients with an age of more than 65 years significantly less frequently knew the meaning of tumor markers (p < 0.001). Fewer women suffering from ovarian cancer were uninformed about tumor markers as compared to women suffering from other malignancies (p < 0.001). CONCLUSION: We conclude that the majority of patients in oncological follow-up are interested in tumor markers and want to be informed about these substances. Periodical serum tumor marker sampling is regarded as a safety measure by patients, but information about tumor markers should be improved. PMID- 10810436 TI - Prognostic benefit of extended radical lymphadenectomy for patients with gastric cancer. AB - BACKGROUND: The therapeutic value of extensive gastric lymphadenectomy in gastric cancer is controversial. We retrospectively investigated the effect of extended lymphadenectomy on survival in 882 patients with gastric cancer. MATERIALS AND METHODS: From 882 patients who underwent gastrectomy, D0 or D1 lymphadenectomy was performed on 137 patients, D2 lymphadenectomy on 524 and D3 lymphadenectomy on 221. Curative gastrectomy was performed on 771 patients and the 5-year survival rate of patients had undergone D0 or D1 lymphadenectomy (D1 group) and that of patients who had undergone D2 lymphadenectomy (D2 group) was compared with that of patients had undergone D3 lymphadenectomy (D3 group). RESULTS: In each stage, the 5-year survival rates of patients who had undergone curative operations (n = 771) were compared among the D1, D2 and D3 groups. The 5-year survival rates were as follows: Stage I: n = 510, 89.3% (D1 group: n = 91, 85.8%; D2 group: n = 384, 90.3%; D3 group: n = 35, 88.1%; p = 0.539), Stage II: n = 83, 77.8% (D2 group: n = 35, 82.7%; D3 group: n = 48, 74.3%; p = 0.601), Stage III: n = 133, 50.2% (D2 group: n = 46, 39%; D3 group: n = 87, 56.1%; p = 0.027), Stage IV: n = 45, 8.9% (D2 group: n = 10, 0%; D3 group: n = 35, 11.4%; p = 0.588). Postoperative complications were detected in 10.4% of the cases and the in hospital mortality rate was 2.4%. The postoperative morbidity rate of the D3 group (15.8%) was significantly higher than that of the other groups (D1 group: 7.3% and D2 group: 9%; p = 0.008). However, the in-hospital mortality rate of the D3 group (1.4%) was not different from that of the other groups (D1 group: 3.7% and D2 group: 2.5%; p = 0.374). CONCLUSION: These results indicate that D3 lymphadenectomy might be performed as safely as D1 or D2 lymphadenectomy on patients with gastric cancer. In addition, D3 lymphadenectomy might provide a survival benefit for patients with Stage III or Stage IV gastric cancer. PMID- 10810437 TI - Accuracy of tissue polypeptide specific antigen (TPS) in the diagnosis of ovarian malignancy. AB - This paper evaluates the usefulness of determining the level of serum Tissue Polypeptide Specific Antigen (TPS) by TPS ELISA in the diagnosis of ovarian malignancies and compares it with the results of histological examination of the ovaries. The study covers 92 patients who had been clinically diagnosed with ovarian tumors by history, physical examination and pelvic examination with or without pelvic ultrasonography. All of them underwent surgical treatment by exploratory laparotomy in Siriraj Hospital between May 1, 1996 and 31 March, 1997. None of the patients had been treated with chemotherapy or hormonal therapy, and in no case were there any previously diagnosed malignancies. TPS was measured in the serum of 92 patients who were preoperatively diagnosed with ovarian tumor: 52 patients had benign pelvic masses and 40 patients had malignant ovarian tumors. Using the criterion for TPS positivity defined by the manufacturer (80 U/L), TPS levels were elevated in 28.8% of benign pelvic mass patients and in 90% of malignant ovarian tumor patients. Statistical analysis using a two-by-two table at every cut-off TPS level and a receiver operating characteristic (ROC) curve, the optimal accuracy, sensitivity, specificity, positive predictive value, negative predictive value, false positive rate and false negative rate were 79.4, 90.0, 71.2, 70.6, 82.9, 28.9 and 10.0%, respectively using the positivity criterion of 80 U/L. We conclude that determination of serum TPS level by TPS ELISA in the diagnosis of ovarian malignancy is good and clinically acceptable. A TPS level greater than 80 U/L is a useful positivity criterion for screening for malignant ovarian tumors, while a TPS level greater than 180 U/L is a positivity criterion for differentiating malignant ovarian tumor from benign pelvic mass. Because of its high false positive rate, any patient with TPS greater than 80 U/L should be further investigated for malignant ovarian tumor. PMID- 10810438 TI - Predictive value of the combined serum CA 125 and TPS during chemotherapy and before second-look laparotomy in epithelial ovarian cancer. AB - CA 125 and TPS levels were measured in the serum of 33 patients with advanced non mucinous epithelial ovarian cancer. These patients were in clinical complete response (CR) after postoperative chemotherapy. The median level of CA 125 decreased rapidly after the first cycle of treatment, and then decreased slowly throughout the rest of the treatment. Patients with a positive second-look had significantly higher CA 125 levels than those with pathological CR. The median level of TPS decreased linearly throughout the treatment. The negative predictive value of the level of CA 125 was 72.7% (95% CI 39.3-92.7) using 10 U/ml as cut off, while that of the level of TPS was 73.3% (95% CI 44.8-91.1), using a cut-off value of 50 U/L. The combined CA 125 and TPS criteria had a better negative predictive value of 88.9% (95% CI 63.9-89.1). PMID- 10810439 TI - Human papillomavirus in head and neck carcinomas: prevalence, physical status and relationship with clinical/pathological parameters. AB - The association of human papillomavirus (HPV) infection to head and neck squamous cell carcinomas was evaluated in 66 patients affected by tumours of the oral cavity (n = 38), the tonsil (n = 4), the pharynx (n = 2), and the larynx (n = 22). HPV DNA was detected by PCR-based assays, recognizing late and early genes. Twenty-four cases were HPV infected (36.4%), mostly by high and/or intermediate risk types. HPV 16 was integrated in 7/12 positive tumours without site specificity. HPV infection was not related to age, gender, tumour stage, differentiation grade, and use of alcohol and/or tobacco. The findings indicate that HPV infection may be related to a proportion of head and neck carcinomas but its association is not as clear as that found in cervical cancer. PMID- 10810440 TI - Serum matrix metalloproteinase-9 level associated with stromal reaction in patients with gastric cancer. AB - BACKGROUND: Matrix metalloproteinase 9 (MMP-9) plays an important role in cancer invasion and metastasis. The relationship between serum MMP-9 levels and clinicopathological factors in gastric cancer patients is not clear. MATERIALS AND METHODS: Preoperative serum was obtained from 170 patients who had undergone gastrectomy for gastric cancer at the Veterans General Hospital-Taipei. The serum MMP-9 level was measured using a sandwich enzyme-linked immunoassay by monoclonal antibodies. RESULTS: The median serum MMP-9 level was 368.6 ng/mL (range: 43.9 1871.3 ng/mL). Univariate analysis showed that stromal reaction and Lauren's histological classification were two factors related to serum MMP-9 levels (p = 0.014 and p = 0.030 respectively). Multivariate analysis revealed that stromal reaction was the only factor independently (odds ratio: 1.695) associated with MMP-9 levels. Patients with a serum MMP-9 level < 368.5 ng/mL had a tendency towards better survival rate (5-year survival rate: 64.4%) than those with > 368.5 ng/mL (58.5%), but this tendency did not reach statistical significance (p = 0.512). CONCLUSION: These data suggests that serum MMP-9 levels are associated with stromal reaction in gastric cancer. PMID- 10810441 TI - Serum matrix metalloproteinases -2, -9 and tissue inhibitors of metalloproteinases -1, -2 in lung cancer--TIMP-1 as a prognostic marker. AB - The immunoreactive protein for the tissue inhibitor of the metalloproteinase (TIMP)-1 and -2 as well as for the matrix metalloproteinase (MMP)-2 and -9 was quantified from the sera/plasma of 90 lung cancer patients and 20 control subjects with enzyme linked immunoassays (ELISA) using specific monoclonal antibodies. Free MMP-2 and that bound to the inhibitor, the MMP-2/TIMP-2 complex were measured separately using different ELISAs. For the detection of MMP-9, TIMP 1 and TIMP-2, the total protein was measured to quantify both free and complex forms. Serum protein levels for TIMP-1, TIMP-2 and the MMP-2/TIMP-2 complex differed significantly in patients with lung cancer when compared to controls. TIMP-1 levels were found to be higher in lung cancer than in controls, whereas TIMP-2 and MMP-2/TIMP-2 complex levels were lower in lung cancer than in the sera of the control subjects. High TIMP-1 (> 300 ng/ml) or MMP-9 (> 30 ng/ml) correlated to poor cumulative survival in lung cancer patients (log rank P < 0.05). High TIMP-1 indicated a poor prognosis, especially in squamous cell cancer and in NSCLC patients with stage III: 66% and 70%, respectively, of the patients with low TIMP-l serum levels survived for more than one year, when only 25% and 20%, respectively, of the patients with high serum levels for TIMP-1 protein survived at that time. 56% of lung cancer patients with a plasma MMP-9 level < 30 ng/ml survived for 12 months when only 31% of the lung cancer patients with high MMP-9 plasma levels survived for more than one year. Also this difference was significant (log rank analysis, P < 0.05). Our results suggest that the factors of the metalloproteinase system might be important in lung cancer progression. TIMP-1 as well as MMP-9 could serve as prognostic markers, and their values could be investigated in the follow-up of lung cancer patients when selecting patients for systemic chemotherapy or other treatment modalities. PMID- 10810442 TI - Vascular endothelial growth factor expression in progression of cervical cancer: correlation with thymidine phosphorylase expression, angiogenesis, tumor cell proliferation, and apoptosis. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) has been linked not only to angiogenic activity but also to thymidine phosphorylase (TP), rapid tumor growth, and inhibition of apoptotic cell death. Our purpose was to examine how VEGF expression affect these factors in cervical cancer at varying stages of progression. METHODS: VEGF expression, TP expression, the microvessel count (reflected by factor VIII-related antigen), and proliferating cell nuclear antigen (PCNA) were assessed immunohistochemically in 19 specimens of normal cervical epithelium, 35 of carcinoma in situ (CIS), 34 of microinvasive carcinoma (MIC), and 34 of invasive cervical squamous cell carcinoma (SCC). Apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. RESULTS: VEGF expression progressively increased along a continuum from normal epithelium to invasive SCC (P < 0.0001). VEGF expression significantly correlated with TP expression and PCNA index (P < 0.01 and P < 0.0001, respectively). In analyses within histological stages, VEGF expression significantly correlated with the PCNA index in CIS and MIC (P < 0.01 and P < 0.05, respectively), but not in invasive SCC. The PCNA index for combined analysis of VEGF and TP expression was similar to that for VEGF expression alone. VEGF expression tended to correlate with microvessel count, however, the difference was not significant (P = 0.09). No significant correlation was observed between VEGF expression and the apoptotic index. CONCLUSIONS: VEGF expression may stimulate tumor cell proliferation in the early stages of cervical cancer, and may be responsible for cervical tumorigenesis. PMID- 10810443 TI - Early effects of adjuvant tamoxifen therapy on serum hormones, proteins and lipids. AB - Tamoxifen is widely used as an adjuvant treatment for breast cancer. To correctly interpret laboratory test results during tamoxifen treatment, clinicians should be aware of the possible effects of the drug on laboratory tests. This study investigated the effects on serum hormones, proteins, lipids and common biochemistry in seven postmenopausal women with breast cancer during 3 months after initiating the therapy. Statistically significant decreases occurred in serum gonadotropins, alkaline phosphatase, calcium, total protein, prealbumin, orosomucoid, haptoglobin, immunoglobin M and total cholesterol whilst significant increases occurred in serum sex hormone-binding globulin (SHBG), cortisol, parathyroid hormone, aspartate aminotransferase, urate, alpha-1-antitrypsin and ceruloplasmin. The alterations could result from tamoxifen therapy, radiation or changes in lifestyle. All the changes, apart from serum urate, remained within the reference limits. In addition, only serum gonadotropins, SHBG, urate and cholesterol showed clinically significant changes. Alterations in the other laboratory tests are unlikely to disturb diagnoses based on laboratory test results during tamoxifen therapy. PMID- 10810444 TI - Biomimetic transport and rational drug delivery. AB - Medicine and pharmaceutics are encountering critical needs and opportunities for transvascular drug delivery that improves site targeting and tissue permeation by mimicking natural tissue addressing and transport mechanisms. This is driven by the accelerated development of genomic agents requiring targeted controlled release. Although rationally designed for in vitro activity, such agents are not highly effective in vivo, due to opsonization and degradation by plasma constituents, and failure to transport across the local vascular endothelium and tissue matrix. A growing knowledge of the addresses of the body can be applied to engineer "Bio-Logically" staged delivery systems with sequential bioaddressins complementary to the discontinuous compartments encountered--termed discontinuum pharmaceutics. Effective tissue targeting is accomplished by leukocytes, bacteria, and viruses. We are increasingly able to mimic their bioaddressins by genomic means. Approaches described in this commentary include: (a) endothelial directed adhesion mediated by oligosaccharides and carbohydrates (e.g. dermatan sulfate as a mimic of sulfated CD44) and peptidomimetics interacting with adhesins, selectins, integrins, hyaluronans, and locally induced growth factors (e.g. vascular endothelial growth factor, VEGF) and coagulation factors (e.g. factor VIII antigen); (b) improved tissue permeation conferred by hydrophilically "cloaked" carrier systems; (c) "uncloaking" by matrix dilution or selective triggering near the target cells; and (d) target binding-internalization by terminally exposed hydrophobic moieties, cationic polymers, and receptor-binding lectins, peptides, or carbohydrates. This commentary also describes intermediate technology solutions (e.g. "hybrid drugs"), and highlights the high-resolution, dynamic magnetic resonance imaging and radiopharmaceutical imaging technologies plus the groups and organizations capable of accelerating these important initiatives. PMID- 10810445 TI - Genetic variation of basal iron status, ferritin and iron regulatory protein in mice: potential for modulation of oxidative stress. AB - Toxic and carcinogenic free radical processes induced by drugs and other chemicals are probably modulated by the participation of available iron. To see whether endogenous iron was genetically variable in normal mice, the common strains C57BL/10ScSn, C57BL/6J, BALB/c, DBA/2, and SWR were examined for major differences in their hepatic non-heme iron contents. Levels in SWR mice were 3- to 5-fold higher than in the two C57BL strains, with intermediate levels in DBA/2 and BALB/c mice. Concentrations in kidney, lung, and especially spleen of SWR mice were also greater than those in C57BL mice. Non-denaturing PAGE of hepatic ferritin from all strains showed a major holoferritin band at approximately 600 kDa, with SWR mice having > 3-fold higher levels than C57BL strains. SDS PAGE showed a band of 22 kDa, mainly representing L-ferritin subunits. A trace of a subunit at 18 kDa was also detected in ferritin from SWR mice. The 18 kDa subunit and a 500 kDa holoferritin from which it originates were observed in all strains after parenteral iron overload, and there was no major variation in ferritin patterns. Although iron uptake studies showed no evidence for differential duodenal absorption between strains to explain the variation in basal iron levels, acquisition of absorbed iron by the liver was significantly higher in SWR mice than C57BL/6J. As with iron and ferritin contents, total iron regulatory protein (IRP-1) binding capacity for mRNA iron responsive element (IRE) and actual IRE/IRP binding in the liver were significantly greater in SWR than C57BL/6J mice. Cytosolic aconitase activity, representing unbound IRP-1, tended to be lower in the former strain. SWR mice were more susceptible than C57BL/10ScSn mice to the toxic action of diquat, which is thought to involve iron catalysis. If extrapolated to humans, the findings could suggest that some people might have the propensity for greater basal hepatic iron stores than others, which might make them more susceptible to iron-catalysed toxicity caused by oxidants. PMID- 10810446 TI - Energy-dependent accumulation of calcium antagonists in catecholamine storage vesicles. AB - The calcium antagonists verapamil, nitrendipine, mibefradil, and amlodipine accumulate in chromaffin granule ghosts with apparent equilibrium partition coefficients [(mol/mg membrane lipid)/(mol/mg solvent water)] of 246 +/- 105 (N = 8), 2700 +/- 600 (N = 4), 7400 +/- 2200 (N = 4), and 8100 +/- 1100 (N = 5), respectively. In the presence of 1.2 mM MgATP, the partition coefficients were 854 +/- 206 (N = 10), 2300 +/- 600 (N = 4), 32,700 +/- 8,900 (N = 7), and 20,300 +/- 5,000 (N = 11) for verapamil, nitrendipine, mibefradil, and amlodipine, respectively. Except for nitrendipine, the apparent partition coefficients in the presence of MgATP were significantly different from the control (P < 0.001). For amlodipine and verapamil, the vacuolar H(+)-ATPase inhibitors bafilomycin A1 (30 nM) and N-ethylmaleimide (2 mM) and the protonophore (uncoupler) carbonyl cyanide m-chlorophenylhydrazone (CCCP, 10 microM) completely blocked the increase in partition coefficients in response to MgATP. The extra amlodipine, mibefradil, and verapamil that accumulated in response to MgATP were released into the medium by CCCP (10 microM) by 18% (N = 5), 30% (N = 5), and 88% (N = 5) for amlodipine, mibefradil, and verapamil, respectively. Thus, amlodipine, mibefradil, and verapamil, but not nitrendipine, accumulate in catecholamine storage vesicles in response to delta mu H+ generated by the endogenous V-type H(+)-ATPase, and are partially released by de-energetisation. Hence, these calcium antagonists can reach unexpectedly high concentrations in certain target cells, and give pharmacodynamic properties not shared by nitrendipine. PMID- 10810447 TI - Ligand-binding affinity of the type 1 and 2 inositol 1,4,5-trisphosphate receptors: effect of the membrane environment. AB - The inositol 1,4,5-trisphosphate (InsP3) receptor is essential for Ca2+ release from intracellular stores. There are three InsP3 receptor types which are targets for several types of regulation. Ca2+, phosphorylation, and protein-protein interactions may contribute to the complex pattern of the Ca2+ signal in stimulated cells. Furthermore, the 3 receptor types could have different affinities for InsP3. We compared the affinities of the type 1 receptor from the cerebellum with the liver type 2 receptor both in their membrane environment and after isolation by immunoprecipitation. Measurements of [3H]InsP3 binding in a cytosol-like medium revealed that the Kd of the liver receptor (45 +/- 5 nM, N = 14) was higher than the Kd of the cerebellar receptor (28 +/- 3 nM, N = 9). Solubilization and immunopurification of the liver InsP3 receptor resulted in a 10-fold increase in its affinity for InsP3. The affinity of the cerebellar receptor did not change under these conditions. Therefore, the extraction of the liver and the cerebellar receptors from their membrane environments induced an inversion of their relative affinities. Treatment of liver membranes with low concentrations of detergents also increased the affinity for InsP3 binding. These data indicate that the type 1 and the type 2 InsP3 receptors have different affinities for InsP3 and that the properties of the type 2 receptor are strongly regulated by hydrophobic interactions within its membrane environment. PMID- 10810448 TI - Maintenance of differential methotrexate toxicity between cells expressing drug resistant and wild-type dihydrofolate reductase activities in the presence of nucleosides through nucleoside transport inhibition. AB - Methotrexate (MTX), a potent inhibitor of dihydrofolate reductase (DHFR), has been used widely as a chemotherapeutic agent and as a selective agent for cells expressing drug-resistant DHFR activity. MTX deprives rapidly dividing cells of reduced folates that are necessary for thymidylate synthesis and de novo purine nucleotide synthesis. However, MTX toxicity can be circumvented by salvaging thymidine (TdR) and purine nucleosides. Here we have investigated conditions under which nucleoside transport inhibition can be used to maintain differential MTX toxicity between unmodified cells and cells expressing drug-resistant DHFR activity in the presence of exogenous nucleosides. PA317 cells (a 3T3 derivative cell line) were rescued from the toxicity of 0.1 microM MTX by 1.0 microM TdR in the presence of 100 microM inosine. The nucleoside transport inhibitor dipyridamole (DP) resensitized these cells to MTX, even in the presence of exogenous nucleosides. Furthermore, PA317 cells transduced with any of three retroviruses encoding drug-resistant DHFRs remained resistant to MTX over all concentrations tested (up to 10.0 microM) in the presence of DP. Similar results were obtained in transduced HuH7 and K562 cell lines, a human hepatoma and a human leukemia cell line, respectively. We conclude that nucleoside transport inhibition increases the toxicity and selectivity of MTX in cultured cells, and therefore is an effective way to maintain differential MTX toxicity between unmodified and DHFR-modified cells. Our results support the use of nucleoside transport inhibition in in vivo selection protocols involving the liver and hematopoietic systems. PMID- 10810450 TI - Inter-species variation in the metabolism and inhibition of N-[(2' dimethylamino)ethyl]acridine-4-carboxamide (DACA) by aldehyde oxidase. AB - N-[(2'-Dimethylamino)ethyl]acridine-4-carboxamide (DACA) is a new anticancer agent currently undergoing clinical trials. The metabolism of DACA to acridone metabolites by aldehyde oxidase (AO) (EC 1.2.3.1) appears to play a major role in its elimination in human patients and rodents. The aim of this study was to compare the ability of human, guinea pig, and rat AO preparations to metabolise DACA, and to determine if either animal model was appropriate for predicting AO mediated DACA-drug interactions in humans. Both human and rodent liver samples were homogenised in buffer before sequential centrifugation to produce the cytosol fraction. Human supernatant underwent an additional ammonium sulphate precipitation procedure, which produced a 2-fold increase in enzyme activity per milligram of protein. After incubations with DACA (range, 0-200 microM), DACA 9(10H)-acridone formation was determined by HPLC analysis. Michaelis-Menten parameters, Km and Vmax, were determined from the best fit curves by nonlinear regression. Three of the four human liver preparations had similar DACA intrinsic clearance values (Vmax/Km) ranging from 0.27 to 0.35 mL/min/mg protein, whereas both the rat and guinea pig had approximately 7- and 160-fold greater intrinsic clearances, due to lower Km values in rats (4.5 +/- 0.7 microM) and guinea pigs (0.15 +/- 0.1 microM) compared with humans (28.3 +/- 8.3 microM, N = 4). Amsacrine, menadione, and 7-hydroxy-DACA were potent inhibitors of DACA metabolism in all three species, but 10-fold differences in IC50 values were apparent between species. In addition, SKF-525A was a potent inhibitor of the metabolism of DACA in rat cytosol but caused minimal inhibition in the guinea pig or human preparations. These results suggest that neither rat nor guinea pig AO preparations are suitable for predicting AO-mediated DACA-drug interactions in humans. PMID- 10810451 TI - Influence of an aziridine precursor on the in vitro binding parameters of rat and ovine corticosteroid-binding globulin (CBG). AB - Aziridines are highly reactive alkylating compounds used in cancer treatment. Salsola tuberculatiformis Botsch., which causes prolonged gestation in sheep and contraception in rats, contains a very labile hydroxy-phenylaziridine or its precursor. A less labile analogue, 2-(4-acetoxyphenyl)-2-chloro-N-methyl ethylammonium chloride (Compound I), was synthesized and has been shown to be contraceptive in rats and to be stabilized by corticosteroid-binding globulin (CBG). The current study compared the binding parameters of rat and ovine CBG and evaluated the effect of the aziridine precursor, Compound I, on these parameters. Kd and Bmax values of 0.646 and 578 nM for corticosterone binding to rat CBG and 0.577 and 19.8 nM for cortisol binding to sheep CBG, respectively, were measured. In competitive binding studies with rat plasma, Ki values of 3.48 nM, 0.856 nM, 22.2 nM, 722 microM, and > 1,000,000 microM for cortisol, corticosterone, progesterone, Compound I, and synephrine (Compound II), respectively, were found, while in sheep plasma the values were 0.409 nM, 1.78 nM, 5.28 nM, 594 microM, and > 1,000,000 microM, respectively. Concentrations of Compound I equivalent to an effective pharmacological dose resulted in a significant (P < 0.01) decrease in CBG bound corticosterone and a significant (P < 0.01) increase in free corticosterone in rat plasma. In sheep, a similar effect was observed with cortisol. Progesterone binding, however, did not appear to be affected significantly by Compound I in either rat or sheep plasma. Compound I was found to be a competitive inhibitor of glucocorticoid binding to CBG. These results suggest that binding of Compound I to CBG with concomitant displacement of endogenous glucocorticoids, but not progesterone, may be part of the mechanism of action of these phenylaziridine compounds. PMID- 10810449 TI - Ebselen as a peroxynitrite scavenger in vitro and ex vivo. AB - We have previously shown that peroxynitrite (PN) selectively impaired prostacyclin (PGI2)-dependent vasorelaxation by tyrosine nitration of PGI2 synthase in an in situ model (Zou MH, Jendral M and Ullrich V, Br J Pharmacol 126: 1283-1292, 1999). By using this established model, we tested whether or not ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one), which reacts rapidly with the anionic form of PN, affected PN inhibition of PGI2 synthase. Administration of ebselen (1 to 50 microM) to bovine coronary strips 5 min prior to PN (1 microM) treatment neither prevented PN-triggered vasoconstriction nor the inhibition of PGI2 release. In line with these results, ebselen affected neither PN inhibition of the conversion of [14C]-PGH2 into 6-keto-PGF1 alpha nor the nitration of PGI2 synthase in bovine aortic microsomes. Following the hypothesis that a reaction of ebselen with cellular thiols could have caused the inefficiency of ebselen, we observed that free ebselen quickly reacted with thiols in both coronary strips and in aortic microsomes to form two metabolites, one of which was identified as the ebselen-glutathione adduct, whereas the other had a similar retention time to that of the ebselen-cysteine adduct. The nitration of phenol by PN in a metal free solution could be blocked more efficiently in the presence of ebselen or glutathione alone than in the presence of both, indicating that like selenomethionine and other selenocompounds, ebselen-thiol adducts were less reactive towards PN than ebselen itself. Further evidence came from the results that ebselen became effective in preventing the inhibition and nitration of PGI2 synthase after thiol groups of microsomal proteins were previously oxidized with Ellman's reagent. We conclude that in cellular systems ebselen is present as thiol adducts and thus loses its high reactivity towards PN, which is required to compete with the nitration of PGI2 synthase. PMID- 10810452 TI - Agonistic and synergistic activity of tamoxifen in a yeast model system. AB - The background of agonist/antagonist behaviour of the non-steroidal antiestrogen tamoxifen is still not fully understood. Depending on cell type, its activities range from full agonistic to antagonistic in different tissues. We investigated the transactivational properties of tamoxifen in a basic yeast model system which reconstitutes ligand-dependent human estrogen receptor-alpha (hER alpha) gene activation. Tamoxifen exerted low agonist activity in this system compared to 17 beta-estradiol (E2). Efficiencies and potencies of several isomers were calculated by fitting experimental data with a logistic dose-response function. Cis-, trans- and cis-transtamoxifen and trans-4-hydroxytamoxifen (4-OHT) showed comparable efficiencies and potencies in yeast. When subeffective doses of trans , cis-/trans-, or trans-4-OH tamoxifen were combined with increasing 17 beta estradiol concentrations, even a synergistic increase in efficiencies could be observed. Interestingly, the cis-isomer did not show this synergistic effect. Mutation of the N-terminus of the estrogen receptor changed the transactivational behaviour of tamoxifen and abolished the synergistic action with 17 beta estradiol. Except for 4-OHT, the potencies of the investigated isomers, defined as ligand concentrations with half-maximal response, highly correlated with the binding affinities to hER alpha. Therefore, cis-, trans-, and cis-/trans tamoxifen could be regarded as full agonists in yeast, while 4-OHT was regarded as a partial antagonist in yeast. Furthermore, these results indicate that the functional difference between trans-tamoxifen and trans-4-OHT is not due to their different affinities for the receptor protein. PMID- 10810453 TI - Differential regulation of cyclooxygenase-2 (COX-2) mRNA stability by interleukin 1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) in human in vitro differentiated macrophages. AB - Cyclooxygenase-2 (COX-2) is a highly inducible gene in macrophages by pro inflammatory cytokines. A major mechanism for cytokine-induced COX-2 expression is stabilization of COX-2 mRNA. In this study, we examined the induction of COX-2 expression by interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF alpha) in human primary in vitro differentiated macrophages. IL-1 beta (5 ng/mL) or TNF-alpha (1 ng/mL) induced up to an approximately 40-fold increase of COX-2 mRNA in macrophages during a 2 to 2.5-hr incubation. Run-off experiments demonstrated that cytokine stimulation had only a mild effect on the COX-2 transcription rate (approximately 10-40% increase). The translation blocker cycloheximide (CHM) (10 mg/mL) superinduced COX-2 mRNA during 2 hr of incubation and further stabilized the COX-2 mRNA (T1/2 > 4 hr). The CHM-superinduced COX-2 mRNA was subject to a rapid degradation after removal of CHM (T1/2 < 1 hr). Both IL-1 beta and TNF-alpha stabilized cytokine-induced COX-2 mRNA (T1/2 > or = 2 hr). Maximal stabilization of COX-2 mRNA after a short-term stimulation required the continued presence of IL-1 beta in the medium. Long-term treatment of TNF alpha destabilized the induced COX-2 mRNA. Cells simultaneously treated with both IL-1 beta and TNF-alpha had a reduced induction of COX-2, IL-1 beta, and IL-6 mRNA. In transcription-arrested cells, the translation blocker puromycin affected the TNF-alpha-induced stabilization and destabilization of COX-2 mRNA, but not the IL-1 beta-induced stabilization. The studies suggest that positive and negative regulation of mRNA stability may play a major role in cytokine-mediated COX-2 induction in human macrophages. TNF-alpha may play both pro-inflammatory and protective roles during inflammation by regulation of pro-inflammatory gene transcripts. PMID- 10810454 TI - Role of apoptosis in cisplatin-induced toxicity in the renal epithelial cell line LLC-PK1. Implication of the functions of apical membranes. AB - The role of apoptosis and the implications of the functions of apical membranes in cisplatin-induced nephrotoxicity were investigated using the kidney epithelial cell line LLC-PK1. When LLC-PK1 cells were treated with 30 microM cisplatin, the number of floating cells was increased markedly. However, the number was not increased by treatment with 1 mM cisplatin, suggesting that different mechanisms were involved in the toxicities of these two treatments. DNA fragmentation, condensation of nuclear chromatin, and the absence of trypan blue staining suggested that cellular toxicity following treatment with 30 microM cisplatin for 24 hr was mediated predominantly by apoptosis. Specific activities of apical enzymes (gamma-glutamyltransferase, EC 2.3.2.2; and alkaline phosphatase, EC 3.1.3.1) in LLC-PK1 cells were decreased markedly by treatment with 30 microM cisplatin for 24 hr, whereas neither lactate dehydrogenase (LDH; EC 1.1.1.27) release nor a decrease in cellular protein content was observed following the same treatment. In addition, concomitant treatment with reduced glutathione completely attenuated both the apoptosis and the decrease of apical enzyme activities induced by 30 microM cisplatin. Neither DNA fragmentation nor condensation of chromatin was induced by treatment with 1 mM cisplatin for 12 hr. However, LDH release and a decrease in cellular protein level were induced by 1 mM cisplatin, suggesting that the toxic effect was due to necrosis. Under these conditions, specific activities of apical enzymes were not decreased. These results suggested that apoptosis was more responsible than necrosis for the loss of apical functions in cisplatin-induced toxicity in LLC-PK1 cells. PMID- 10810455 TI - Role of nitric oxide in lipopolysaccharide-induced oxidant stress in the rat kidney. AB - Lipopolysaccharide (LPS)-induced renal oxidant injury and the role of nitric oxide (NO) were evaluated using the inducible nitric oxide synthase (iNOS) inhibitor L-iminoethyl-lysine (L-NIL). One group of male rats received LPS (Salmonella minnesota; 2 mg/kg, i.v.). A second group received LPS plus L-NIL (3 mg/kg, i.p.). A third group received saline i.v. At 6 hr, iNOS protein was induced in the kidney cortex, and plasma nitrate/nitrite levels were increased from 4 +/- 2 nmol/mL in the Saline group to 431 +/- 23 nmol/mL in the LPS group. The value for the LPS + L-NIL group was reduced significantly to 42 +/- 9 nmol/mL. LPS increased blood urea nitrogen levels from 13 +/- 1 to 47 +/- 3 mg/dL. LPS + L-NIL reduced these levels significantly to 29 +/- 2 mg/dL. Plasma creatinine levels were unchanged in all groups. Tissue lipid peroxidation products in the kidney were increased from 0.16 +/- 0.01 nmol/mg in the Saline group to 0.30 +/- 0.03 nmol/mg in the LPS group. LPS + L-NIL reduced the values significantly to 0.22 +/- 0.02 nmol/mg. Intracellular glutathione levels were decreased in the kidneys from 1.32 +/- 0.1 nmol/mg in the Saline group to 0.66 +/ 0.08 nmol/mg in the LPS group. LPS + L-NIL increased the levels significantly to 0.99 +/- 0.13 nmol/mg. LPS increased the 3-nitrotyrosine-protein adducts in renal tubules as detected by immunohistochemistry, indicating the generation of peroxynitrite. L-NIL decreased adduct formation. These data indicated that LPS induced NO generation resulted in peroxynitrite formation and oxidant stress in the kidney and that inhibitors of iNOS may offer protection against LPS-induced renal toxicity. PMID- 10810456 TI - Treatment of bone tumors by total excision and replacement with massive autologous and homologous grafts. 1966. PMID- 10810457 TI - P-glycoprotein levels predict poor outcome in patients with osteosarcoma. AB - To evaluate the relationship between the expression of P-glycoprotein by osteosarcomas and the rate of metastasis and death, a retrospective review of 172 patients who were diagnosed with osteosarcoma between 1987 and 1992 was performed. Forty patients had P-glycoprotein levels available. The majority of the osteosarcomas were Stage II-B (33 patients), with the remaining seven being Stage III. Tumor sites included 25 femurs, seven humeri, five tibias, and one each of pelvis, radius, and fibula. The patients with Stage III disease at presentation were treated differently from the time of diagnosis and therefore, these seven patients with Stage III osteosarcoma were excluded from additional analyses. The expression of P-glycoprotein by cultured tumor cells from biopsy specimens was determined using immunofluorescent microscopy. In the 33 patients with Stage IIB osteosarcoma with detectable P-glycoprotein, 67% (10 of 15) had metastases develop as compared with 28% (five of 18) of patients with undetectable P-glycoprotein. Similarly, 53% (eight of 15) of patients with tumors expressing P-glycoprotein died of disease compared with 11% (two of 18) with no detectable P-glycoprotein. Expression of P-glycoprotein by tumor cells seems to be associated with an estimated ninefold increase in the odds of death and a fivefold increase in the odds of metastases in patients with Stage IIB osteosarcoma. Kaplan-Meier survivorship analysis revealed that patients with detectable P-glycoprotein fared worse in terms of survival time and metastasis free survival. Adjusting for covariates in the Cox proportional hazards model, expression of P-glycoprotein and its level were significantly predictive of time to death in patients with Stage IIB osteosarcoma. PMID- 10810458 TI - Synovial sarcoma. Large size predicts poor outcome. AB - A consecutive series of 38 patients with synovial sarcoma diagnosed and treated in a consistent fashion from 1976 to 1994 was reviewed for prognostic variables. The histologic specimens were reviewed and confirmed by one pathologist. There was a minimum 4-year followup for all surviving patients and no patients were lost to followup. The treatment protocol consisted of surgical excision with a wide or radical margin and limb preservation when possible. In those patients in whom the surgical margin was undefined or was less than a wide margin, perioperative radiation therapy was used. Four patients presented with metastatic disease and all died of their disease. Thirty-four patients had localized disease at presentation. Variables considered in stratifying outcomes included histologic grade, histologic subtype, surgical margin, presence or absence of local recurrence, age, and size of tumor. Of the 34 patients without metastasis there was a strong statistical association between size of tumor and survival: 17 patients with tumors less than 5 cm indiameter had a 100% survival, 12 patients with tumors 5 cm to 10 cm had a 75% survival, and five patients with tumors greater than 10 cm had a 20% survival. The authors urge that a multicenter trial for neoadjuvant chemotherapy be initiated for patients presenting with a synovial sarcoma greater than 10 cm in diameter. PMID- 10810459 TI - Targeting tumor specific translocations in sarcomas in pediatric patients for immunotherapy. AB - In an effort to develop more effective therapies for various sarcomas in pediatric patients, the authors have focused on using recurrent tumor-specific translocations as potential novel tumor antigens. In general, these translocations generate fusion transcription factors. Because cytotoxic T cell lymphocyte receptors recognize peptide fragments bound to major histocompatibility complex Class 1 molecules, it is possible that unique peptides spanning the translocation breakpoint region may be processed, bound to major histocompatibility complex Class I molecules and displayed on the tumor cell surface where they could be susceptible to cytotoxic T cell lymphocyte killing. The authors have investigated the PAX-3-FKHR fusion product seen in alveolar rhabdomyosarcoma, and the EWS-FLI-1 fusion product seen in Ewing's sarcoma. Peptides spanning these fusion regions contain potential major histocompatibility complex Class 1 and Class II binding motifs suggesting they may serve as novel T cell antigens. Preliminary mouse experiments suggest that cytotoxic T cell lymphocytes specific for the PAX-3-FKHR fusion peptide can be generated and can recognize and kill tumor cells bearing the PAX-3-FKHR fusion protein. Clinical trials are ongoing to determine whether this approach will be useful. PMID- 10810460 TI - Epiphyseal plate involvement in osteosarcoma. AB - Fourteen boys (56%) and 11 girls (44%) 4 to 17 years of age (mean, 12.2 years) who had osteosarcoma and open epiphyseal plates were studied. A possible correlation between transepiphyseal spread of osteosarcoma and radiologic and histopathologic findings was investigated. Epiphyseal plate invasion was detected radiologically in only 11 patients (44%), whereas histopathologic examination showed transepiphyseal extension in 21 patients (84%). The authors conclude that the epiphyseal plate is not a barrier against tumor growth and strongly recommend that limb salvage surgery preserving the epiphysis be planned carefully. PMID- 10810461 TI - Osteosarcoma in preadolescent patients. AB - The medical records of boys younger than 11 years and girls younger than 10 years of age with osteosarcoma of the pelvis or extremity were reviewed. Thirty patients were identified who were newly diagnosed but untreated for osteosarcoma. None of these patients had pulmonary metastases. The same four protocols were used to treat the patients in the current study as were used to treat adolescents. The event-free and overall survival was calculated and prognostic factors were assessed. The median followup time was 8 years (range, 6-14 years). The results were compared with the results of older patients treated with the same protocols and with published results. Fourteen patients had pulmonary metastases (47%); among these patients, four also had skeletal metastases (in two of the latter, skeletal metastases appeared before the pulmonary metastases). Event-free survival was 53% and overall survival was 57%. This result is comparable with current survival results in adolescent and older patients. Serum alkaline phosphatase and serum lactic dehydrogenase levels before treatment, height percentile greater than 50%, chemotherapy-induced tumor necrosis, surgical procedure, tumor site, tumor histologic features, and patient gender were not prognostic indicators. The prognosis for prepubertal patients with osteosarcoma is similar to the prognosis of their adolescent and older counterparts. There does not seem to be any indication to treat preadolescent patients with osteosarcoma using alternate therapies. PMID- 10810462 TI - Expandable endoprosthesis reconstruction in skeletally immature patients with tumors. AB - Between September 1984 and January 1996, 32 expandable endoprostheses were used for limb reconstruction after resection of malignant bone tumors in patients who were skeletally immature. The 20 boys and 12 girls ranged in age from 3 to 15 years (mean, 9.7 years). One patient had a Stage IIA tumor, 22 patients had Stage IIB tumors, and seven patients had Stage III tumors according to the classification of the Musculoskeletal Tumor Society. There also were two patients with parosteal osteosarcomas. The histologic diagnosis was osteosarcoma in 23 patients and Ewing's sarcoma in nine. All patients except the patients with parosteal osteosarcoma received standard neoadjuvant therapy. Twenty-two Lewis Expandable Adjustable Prostheses, four modular Wright Medical prostheses, four modular Howmedica prostheses, and two Techmedica expandable prostheses were used. Thirteen patients died, two have no evidence of disease, and 17 are continuously disease free. Sixteen of 32 patients (50%) have not had an expansion procedure because of early death in 10 and early amputation in three. Three patients are waiting to undergo an expansion procedure. Sixteen of the 32 patients (50%) have undergone 32 expansion procedures, to a maximum of 9 cm, without any infection. To maintain range of motion before the expansion procedure, a complete resection of the pseudocapsule was done routinely. Fourteen of the 32 patients did not have complications. Eighteen of the 32 patients had 27 complications. All Lewis Expandable Adjustable Prosthesis endoprostheses and the two nonmodular Techmedica prostheses were associated with a large amount of titanium debris. The children's functional results were similar to the results reported for adults with an average Musculoskeletal Tumor Society rating of good to excellent at the knee, fair to good at the hip, and fair about the shoulder. Rehabilitation of the knee in very young patients (5-8 years) remains problematic and careful selection of patient and family is necessary. The Lewis Expandable Adjustable Prosthesis probably should be reserved for very young patients (5-8 years) and modular systems should be used for large preadolescent and adolescent children. PMID- 10810463 TI - Total knee arthroplasty in patients who have massive osteoarticular allografts. AB - The authors treated 24 patients with total knee arthroplasty who had a massive allograft used to reconstruct the knee and who later had instability, degeneration, or a fracture near the articular surface of the graft develop. Patients then were followed up for a minimum of 2 years and a mean of 8.2 years. Overall, 96% of the patients retained a functional limb, although 46% underwent revision surgery, and an additional 12% had some other major complication. Statistical analysis showed a significant negative effect of chemotherapy on revision-free survival of the prosthesis. Patients with high-grade tumors were at significantly greater risk of fracture of the allograft-prosthesis composite. Certain technical factors were identified that seemed to predispose the allograft prosthesis reconstructions to early failure. Total knee arthroplasty can be used to treat patients with complications of massive osteoarticular allografts and may prolong the functional life of an otherwise successful limb salvage reconstruction. PMID- 10810464 TI - Survivorship and radiographic analysis of knee osteoarticular allografts. AB - An evaluation of long-term survival and radiographic deterioration of 118 knee osteoarticular allografts in 114 patients was performed. Radiographic analysis was done according to the Musculoskeletal Tumor Society grading system. A failure was defined as when the allograft was removed during a revision procedure or amputation. Ten patients were lost to followup during the first 2 years after surgery. Eighteen patients without allograft failures died of complications related to the tumor. Twenty-six allografts failed because of infection (13 allografts), local recurrence (eight allografts), massive resorptions (three allografts) and fractures (two allografts). Sixty-four allografts still were in place at a mean of 98 months (range, 36-360 months) after implantation. The Kaplan-Meier 5-year survival rate for the knee osteoarticular allografts was 73% and the limb preservation rate was 93%. The mean radiographic score was 83%. Sixty-four percent of the allografts showed no radiologic changes or minor articular deterioration. Fourteen percent had narrowing of the joint space of more than 2 mm, and 22% had some form of subchondral bone collapse. Five patients required joint resurfacing to preserve the original allograft. Most of the allograft failures occurred during the first 4 years, and the allograft survival rate for the current series remained unchanged after 5 years. PMID- 10810465 TI - Fine needle aspiration biopsy of primary bone tumors. AB - A review of 66 consecutive fine needle aspiration biopsies of primary bone tumors revealed that 48 (73%) were diagnostic. Twelve (18%) yielded inadequate specimens unsatisfactory for diagnosis, and five (8%) yielded specimens adequate for partial diagnosis. The only error, presumably attributable to sampling error, was an unappreciated dedifferentiated osteosarcoma arising in an otherwise typical giant cell tumor. Fine needle aspiration biopsy obviated the need for open biopsy in 24 patients and simplified surgery in an additional 24 patients by establishing the diagnosis before surgical intervention. A solitary soft tissue recurrence of a giant cell tumor has been the only local recurrence. A review of 26 consecutive patients with osteosarcoma revealed that seven tumors were diagnosed by primary open biopsy. Nineteen patients had fine needle aspiration biopsy, of which 15 were diagnostic and four required supplemental open biopsy. The elapsed time between the initial office visit and the diagnostic confirmation averaged 5 days for patients requiring open biopsy compared with 0 days for patients whose fine needle aspiration biopsy was diagnostic. The total estimated charge for fine needle aspiration biopsy of a distal femoral osteosarcoma was $1060.00 compared with $4312.25 for open biopsy. There have been no local recurrences in patients in either group. Fine needle aspiration biopsy provides an accurate, safe, efficient, well tolerated, and cost-effective method for diagnosing classic primary bone tumors, including osteosarcoma. PMID- 10810466 TI - Primary fibrosarcoma of bone. Outcome after primary surgical treatment. AB - To investigate outcome and evaluate prognostic factors in primary fibrosarcoma of bone, all patients at the authors' institution who had surgical treatment for primary fibrosarcoma of bone from 1910 to 1995 were studied. Medical records, surgical reports, radiographs, and histologic slides of 92 patients (51 males, 41 females; mean age, 38 years; range, 8-84 years) were reviewed. The most common tumor locations were the femur (28 patients), tibia (21 patients), and pelvis (14 patients). Sixty-one tumors (66%) were Enneking Stage IIB. Twenty-nine patients (31.5%) had adjuvant therapy: 16 had radiation, nine had chemotherapy alone, and four had radiation and chemotherapy. Amputation or disarticulation was performed in 61 patients (66%), wide excision was performed in 13 (14%), marginal excision was performed in 15 (16%), and intralesional excision was performed in three (3.3%). Local recurrence occurred in 14 patients (15%) at a median time of 7 months (range, 3-21 months). Metastases developed in 58 of the 85 patients (68%) with Stage I or II tumors at presentation, at a median of 9 months (range, 1-51 months). Survivorship analysis showed that the overall probability of survival was 33.4% at 5 years after surgery. Multivariate analysis showed that the main prognostic risk factors affecting overall survival included age older than 40 years, tumor location in the axial skeleton, and high-grade tumor (Grade 3 or 4). With the high incidence of systemic failure after surgical treatment, perioperative adjuvant treatment modalities should be considered. PMID- 10810467 TI - Review of cellular mechanisms of tumor osteolysis. AB - The cellular and biochemical mechanisms that direct the destruction of bone at sites of tumor osteolysis are unknown. To better understand the mechanisms through which tumors direct bone resorption, research has focused on developing in vivo and in vitro experimental models that are useful for studying this process. In vivo experimental systems have been developed that permit study of tumor osteolysis from human and murine tumors, and that permit the study of tumors that arise from (sarcoma) or can metastasize (breast cancer) to bone. Recent research has focused on three questions: (1) Are osteoclasts or tumor cells responsible for bone resorption during tumor osteolysis? (2) What are the cellular mechanisms that are responsible for bone resorption during tumor osteolysis, and (3) what are the tumor cell products that regulate the cellular mechanisms that are responsible for tumor osteolysis? It has been determined that osteoclasts are responsible for bone resorption at sites of tumor osteolysis by enhancing the binding of osteoclast to bone, by inducing osteoclastic bone resorption, and by stimulating osteoclast formation. Attempts to identify tumor cell products that regulate these cellular mechanisms are in progress, and findings suggest that production of macrophage colony stimulating factor may be required for tumor osteolysis to occur with some tumors. PMID- 10810468 TI - Percutaneous radiofrequency ablation of osteoid osteoma. AB - Osteoid osteoma is a benign bone tumor. Patients usually require surgical treatment for reliable pain relief. Difficulties with intraoperative localization of the tumor and anatomic locations that carry a high morbidity with en bloc resection complicate open surgery. Various methods have been developed to lessen the invasiveness of surgery including computed tomography-guided percutaneous radiofrequency thermal ablation. Eleven patients in three different centers were evaluated and diagnosed with osteoid osteoma based on typical histories, physical examinations, and imaging studies. All patients were treated with computed tomography-guided percutaneous radiofrequency thermal ablation after medical treatment failed. Excellent pain relief was reported in 10 patients. One patient suffered recurrence of a femoral neck lesion despite an initial 7-month period without pain. Patients were given a questionnaire to quantify the effectiveness of percutaneous radiofrequency ablation in terms of pain relief and return to function. The current study shows that percutaneous radiofrequency thermal ablation provides reliable, excellent pain relief and early return to function with minimal morbidity as compared with traditional open techniques. The authors suggest that this technique be used for all patients with extraspinal osteoid osteomas that are not immediately adjacent to neurovascular structures. PMID- 10810469 TI - Quantitative analysis of neural distribution in human coracoacromial ligaments. AB - This study investigated sensory nerve distribution in 27 human coracoacromial ligaments by immunohistochemical methods using antiprotein gene product 9.5 antibody and anticalcitonin gene related peptide antibody. Mean nerve densities were compared among three areas (acromion side, center, and coracoid side) in two groups (patients with rotator cuff tears and patients with shoulder dislocations). In all three areas of both groups, many nerve fibers immunoreactive to antiprotein gene product 9.5 antibody were observed in the periligamentous bursal tissue. However, in the ligament parenchyma, nerve fibers immunoreactive to antiprotein gene product 9.5 antibody were recognized only around blood vessels. Nerve fibers immunoreactive to anticalcitonin gene related peptide antibody were recognized in the periligamentous bursal tissue. However, in the ligament parenchyma, there were no nerve fibers immunoreactive to anticalcitonin gene related peptide antibody. Nerve density of the rotator cuff tear group, as revealed by both immunostainings, showed a significant increase compared with that of the shoulder dislocation group in all three areas. The results of this study show that it is possible the increase in sensory nociceptive nerve fibers in the coracoacromial ligaments may be one of the causes for pain in patients with rotator cuff tears. PMID- 10810471 TI - Atraumatic osteonecrosis of the adult elbow. AB - Osteonecrosis is a disease that leads to joint destruction and often involves large joints, such as the hips, knees, and shoulders. Nontraumatic osteonecrosis of the adult elbow, to the best of the authors' knowledge, has not been reported. Nine adult patients with atraumatic osteonecrosis of 11 elbows were identified. The mean age at presentation was 36 years (range, 26-63 years); five patients were women and four were men. Six elbows involved the capitellum, three involved the lateral epicondyles, one involved the trochlea and radial head, and one involved medial and lateral epicondylar disease. All patients were receiving corticosteroid therapy, and no relationship between the duration or the amount of corticosteroid use and the severity of the osteonecrosis was found. Seven patients with radiographic Stage I and Stage II disease responded well to nonoperative treatments consisting of activity modification, analgesics, and a brief period of immobilization. Nonoperative treatment failed in two patients with Stage III disease, and they had core decompressions for pain relief. One patient with late Stage III disease in both elbows underwent bilateral total elbow arthroplasties. In contrast to the pediatric population, osteonecrosis of the adult elbow potentially can lead to end stage arthritis. If the osteonecrosis is diagnosed early, nonoperative treatment may be effective in relieving pain, although the long-term results of these treatments remain unknown. PMID- 10810470 TI - Glenohumeral osteoarthrosis after the Eden-Hybbinette procedure. AB - Thirty-six patients (36 shoulders) who underwent the Eden-Hybbinette procedure for recurrent anterior dislocation of the shoulder with an average followup of 15 years were evaluated. Evaluation consisted of radiographic assessment in a true anteroposterior view and an axillary lateral view of both shoulders, physical examination, and a questionnaire. Mild glenohumeral osteoarthrosis was present in 1/3 of the patients, and moderate and severe osteoarthrosis was evident in 1/2. There were no signs of osteoarthrosis in four shoulders. Function, as assessed by the Rowe score as modified by Young and Rockwood, was excellent or good in 27 shoulders and fair or poor in nine shoulders. The extent of osteoarthrosis was related to restriction of external rotation, length of followup, and function. PMID- 10810472 TI - Transverse divergent dislocation of the elbow. AB - A girl 6 years 5 months old sustained a rare transverse (mediolateral) divergent dislocation of the elbow and was treated by closed reduction. The arm was immobilized in a cast. Followup was 7 years 5 months. Five years after injury, the patient had recurrent popping on the lateral aspect of the injured elbow during active flexion from complete extension and tenderness on the anterior aspect of the radiohumeral joint. To determine the exact anatomic relation of the proximal radius and ulna to each other and to the distal humerus after transverse dislocation, a skeleton of the elbow was assembled in the dislocated position, and multiple radiographs and photographs were taken and compared with the radiographs of the patient's injured elbow. This showed that the ulna was displaced posteromedially, with the coronoid process impinged against the distal and medial portions of the trochlea, and that the radial head was displaced lateral to the capitellum, anterior and distal to the lateral epicondyle. The distal end of the humerus was caught by the radius and the ulna and was rotated anteriorly and laterally, primarily by the displaced coronoid process. Because of this rotation of the humerus, the lateral radiograph of the elbow gave the impression that the head of the radius was displaced posteriorly. PMID- 10810473 TI - Total hip arthroplasty in young patients. 8- to 13-year results using an uncemented stem. AB - One hundred eight uncemented total hip arthroplasties were performed in 91 patients who were 50 years of age or younger using the Taperloc femoral component. The average age of the patients at time of surgery was 37 years (range, 20-50 years). The mean followup was 10.2 years (range, 8-13 years). No patient was lost to followup. Seven patients (eight hips) died before obtaining the minimum time of 8 years for inclusion in this study. All seven died with their femoral components in place. Of the remaining 100 total hip arthroplasties, no femoral component required revision for aseptic loosening. One femoral component was revised to correct a leg length discrepancy, and one well-fixed femoral component was revised for sepsis. In the 98 total hip arthroplasties that had not undergone femoral component revision, complete radiographic and clinical followup was obtained. Radiographically, 96 (98%) femoral components were determined to have fixation by bone ingrowth, two (2%) femoral components showed stable fibrous ingrowth, and no femoral component was unstable. Femoral cortical osteolysis occurred in seven (7%) hips; major lysis was present in only one (1%). Clinically, 91 (93%) total hip arthroplasties were rated good or excellent; six (6%) were rated fair, and one (1%) was rated poor. Thirty-nine patients with 47 total hip arthroplasties (48%) were engaged in moderate to strenuous manual labor. These results indicate that excellent fixation and minimal lysis can be achieved with an uncemented femoral component in young and active patients at 10 years. PMID- 10810474 TI - Stability and bone preservation in custom designed revision hip stems. AB - Three types of uncemented femoral stems were designed for patients having revision hip surgery, with the goals of promoting axial stability and preserving proximal bone stock. These stems were made individually using computer design and manufacturing technology. Various design features were examined using nonlinear finite element analysis. All stems had lateral, medial, and anterior flares in the proximal region, proximal hydroxyapatite coating, and a collar. Based on a published classification system, the three designs were found suitable for variously encountered cavitary defects. For cases involving small amounts of bone destruction, a primary type of stem was used. With severe cases, an extended polished stem was used. For the worst cases, an extended stem with longitudinal cutting flutes and complete hydroxyapatite coating was necessary. The axial migration was measured radiographically for a 2-year period. The migration rates were comparable with those seen in cemented primary and in custom primary hydroxyapatite coated stems. Dual energy x-ray absorptiometry data were obtained during a 4-year postoperative period. Average bone density in all regions was maintained within 12% of the immediate postoperative values. It was concluded that the proposed system for treating patients needing revision hip surgery showed desirable properties that were comparable to primary hip replacements. PMID- 10810475 TI - Strength and function before and after anterior cruciate ligament reconstruction. AB - Rehabilitation of patients with anterior cruciate ligament injury is a topic of interest among clinicians. Although numerous studies report the deficits after anterior cruciate ligament reconstruction, there are no data available on the changes in strength and functional performance before and 6 months after reconstruction surgery. This is when most patients are returning to sports activities. In the current study 31 patients with complete unilateral anterior cruciate ligament ruptures followed a controlled rehabilitation program emphasizing early range of motion after surgery and quadriceps strengthening before and after undergoing bone-patellar tendon-bone reconstruction. Strength of the hamstrings and quadriceps and five functional activities were assessed at 1 week before and 6 months after surgery. Preoperative strength assessments using a Cybex II dynamometer showed a 12% quadriceps deficit at 60 degrees per second and 9% deficit at 120 degrees per second with no hamstring deficit in the injured leg compared with the uninjured leg. Despite intensive quadriceps training, the postoperative quadriceps deficit increased to 28% at 60 degrees per second and 22% at 120 degrees per second in comparison with the uninjured leg. There was no change in hamstring strength in the injured leg. Most functional measures improved after surgery. The shuttle run improved by 10%, the side step test results improved by 17%, and the carioca test results improved by 23%. No significant difference was found in the results of the hop test. This study highlights the extent of the quadriceps deficits despite functional improvement and the difficulty in restoring quadriceps strength after bone-patellar tendon bone reconstruction. It also questions the relationship of current functional tests to quadriceps strength and the role of maximum strength testing as a predictor of function and of safe sporting performance. PMID- 10810476 TI - Comparison of the single assessment numeric evaluation method and the Lysholm score. AB - The purpose of this study was to determine the correlation between the Lysholm knee score and the Single Assessment Numeric Evaluation method. Between March 1995 and December 1996, 201 followup examinations were performed on 130 college age patients who had undergone anterior cruciate ligament reconstructions. These 201 examinations were divided into five routine followup categories: 3 months, 6 months, 1 year, 2 years, and greater than 2 years. The Lysholm score from each patient's followup questionnaire was correlated with his or her Single Assessment Numeric Evaluation rating, as determined by his or her written response to the following question: On a scale from zero to 100, how would you rate your knee today (100 being normal)? All data were gathered prospectively. The mean Lysholm scores and Single Assessment Numeric Evaluation ratings ranged from 84.0 to 93.4 points and 80.0 to 93.3 points, respectively. The correlation coefficients between the Lysholm scores and the Single Assessment Numeric Evaluation ratings ranged from 0.58 to 0.87 points. The results of this study indicate that Single Assessment Numeric Evaluation ratings correlate well with measured Lysholm scores in patients with anterior cruciate ligament reconstruction. The Single Assessment Numeric Evaluation provides clinicians with an alternative mechanism to gather outcomes data with little demand on time and resources. PMID- 10810477 TI - Ewing's sarcoma of the pelvis. Long-term survival and functional outcome. AB - Fifty patients with Ewing's sarcoma of the pelvis were treated using a multidisciplinary approach; followup of surviving patients averaged 137 months (range, 40-276 months). The addition of surgical resection to the multidisciplinary treatment for all patients was associated with improved survival compared with survival of patients treated with chemotherapy and radiation therapy alone; the addition of surgery to the treatment regimen of 37 patients without metastases also was associated with improved survival. There were no significant differences between the surgical and nonsurgical groups in terms of tumor size, stage of disease, patient age, duration of symptoms before diagnosis, or anatomic site. Surgery was used more often in recently treated patients, but the year of diagnosis and treatment did not significantly affect overall survival, secondary to large confidence intervals. The Short Form-36 and the Musculoskeletal Tumor Society functional evaluation instruments showed a superior level of function in the nonsurgical group, but this difference was not statistically significant. There have been many advances in the treatment of patients with Ewing's sarcoma during the past 3 decades, resulting in improved survival for patients with Ewing's sarcoma of the pelvis. The addition of surgery significantly improved survival and did not show a significant difference in functional outcome. PMID- 10810478 TI - Liposarcoma associated with multiple intramuscular lipomas. A case report. AB - A 71-year-old slender, previously healthy man was admitted to the authors' institution because of a huge painless mass in his left scapular area. Physical examination revealed a second soft tissue mass in his right scapular region and a third soft tissue mass in the anterior side of his right shoulder. Surgical treatment including marginal resection of the second and third small masses followed by wide local resection of the huge tumor was performed. Histologic examination showed that the first mass was a well differentiated lipomalike liposarcoma, whereas the second and third lesions were identified as intramuscular lipomas. A review of the literature showed two cases of retroperitoneal liposarcoma associated with multiple subcutaneous lipomas and two cases of liposarcoma involving an extremity associated with multiple subcutaneous lipomas. There is no previous report in which intramuscular liposarcoma was associated with multiple intramuscular lipomas. PMID- 10810479 TI - Clinical outcome of 38 patients with juxtacortical osteosarcoma. PMID- 10810480 TI - Retrograde nailing of femur fractures in patients with myelopathy and who are nonambulatory. AB - The authors studied 10 consecutive patients with closed femoral shaft or supracondylar fractures who were nonambulatory and who were treated by reamed retrograde intramedullary nailing via an intercondylar notch approach. The study consisted of five women and five men with an average age of 60.7 years (range, 40 89 years). Six patients had spinal cord lesions, one had a brain injury, one had cerebral palsy, one had multiple sclerosis, and one had progressive myelopathy. Three fractures were supracondylar, and seven fractures involved the mid-distal diaphysis. The average time of surgery was 110 minutes (range, 70-225 minutes) with an average estimated blood loss of 288 mL (range, 150-400 mL). There were two postoperative deaths (at 15 days and 2 months, respectively) after the procedure that were attributable to pneumonia. The remaining eight patients were observed for an average of 13 months (range, 6-20 months) after surgery. All fractures healed as evaluated radiographically. Retrograde intramedullary nailing is a simple, safe, and effective alternative to nonoperative treatment for femoral shaft or supracondylar fractures in patients who are nonambulatory. Stabilization by this method allows fracture healing and rapid return of patients to their previous level of function. There were no nonunions, malunions, significant shortening, implant failure, or wound infections. PMID- 10810481 TI - Stress fractures in the Israeli defense forces from 1995 to 1996. AB - This study, encompassing 2591 Israeli soldiers, characterized Israeli soldiers with stress fractures to profile individuals who are prone to experience stress fractures: 318 with clinically and scintigraphically proven high grade stress fractures; 237 soldiers with symptoms but with normal scintigraphy; and 2036 soldiers with no symptoms. Soldiers with high grade stress fractures weighed less (68.4 +/- 7.9 kg versus 70.5 +/- 12.4 kg), smoked less, and reported fewer previous stress fractures, had fewer reports of stress fractures in their family histories, and had fewer incidences of bone diseases than did control subjects. Serum levels of bone specific alkaline phosphatase and osteocalcin were elevated in patients with high grade stress fractures compared with control subjects with no symptoms: 37.6 versus 26.2 units/L, and 10.8 versus 8.8 ng/mL, respectively. Levels of 25-hydroxy vitamin D were lower in patients with high grade stress fractures (25.3 ng/mL) than in control subjects (29.8 ng/mL). This study revealed that several parameters can distinguish soldiers with high grade stress fractures, but their predictive value and precise pathogenetic role remain unclear. PMID- 10810482 TI - Interleukin-10 release and monocyte human leukocyte antigen-DR expression during femoral nailing. AB - This study determined the effect of femoral nailing on the expression of monocyte Class II antigens and interleukin-10 release and sought to differentiate any differences in the release of these elements of immune reactivity in patients undergoing reamed and unreamed nailing. Thirty-two patients presenting with an acute femoral fracture were studied. In 15 patients, the femoral fracture was stabilized with a reamed technique and in 17 patients with an unreamed technique. Venous blood samples were taken at presentation, at anesthetic induction, immediately after nail insertion, and subsequently at 1, 4, and 24 hours and at 3, 5, and 7 days after surgery. Serum interleukin-10 was measured by an enzyme linked immunosorbent assay, and monocyte human leukocyte antigen-DR expression was quantified by flow cytometry. Serum interleukin-10 release and human leukocyte antigen-DR expression on monocytes showed a clear response to the nailing procedure. The group of patients undergoing a reamed femoral nailing procedure showed significantly higher interleukin-10 release and a significant depression in the expression of human leukocyte antigen-DR on monocytes compared with those whose nail had been inserted unreamed. One patient in the reamed femoral nailing group died of adult respiratory distress syndrome 3 days after injury. Reamed intramedullary nailing appears to be associated with greater impairment of immune reactivity than is the unreamed nailing technique. PMID- 10810483 TI - Sequential indium-labeled leukocyte and bone scans to diagnose prosthetic joint infection. AB - Previous studies suggest that sequential technetium-99-hydroxymethyl diphosphonate bone scanning and indium-111 leukocyte scintigraphy may play a role during revision arthroplasty. Preoperative sequential imaging was compared with joint aspiration and clinical assessment during revision knee or hip arthroplasty. Scans were considered positive if indium-111 leukocyte uptake was incongruent or focally more intense than that of technetium-99-hydroxymethyl diphosphonate uptake. Of 166 cases, 22 were infected. Sequential technetium-99 hydroxymethyl diphosphonate and indium-111 leukocyte imaging was 64% sensitive and 78% specific. Fever, physical findings, or sedimentation rate did not identify infection reliably, and preoperative aspirate culture was only 28% sensitive. Positive scintigraphy increased the likelihood of finding infection intraoperatively from 14% to 30%, although negative scintigraphy decreased this likelihood to 7%. Based on the current study, the routine use of sequential technetium-99-hydroxymethyl diphosphonate and indium-111 leukocyte imaging cannot be advocated for differentiating occult infection from mechanical failure in painful, loose total joint arthroplasties. PMID- 10810484 TI - Delayed paraplegia caused by the gradual collapse of an infected vertebra. AB - A case of delayed paraplegia caused by a gradual and progressive collapse of a vertebra after healing of pyogenic spondylitis is reported. A 73-year-old man was treated for a hematogenously seeded pyogenic spondylitis of the first lumbar vertebra. Magnetic resonance imaging showed a high signal intensity of the involved vertebra and adjacent discs and a paravertebral abscess without disc space narrowing. Eight months after healing of the infection, the patient had muscle weakness and paresthesia of the lower extremities, which gradually increased. The plane radiographs revealed a kyphotic deformity of 36 degrees with a collapse of the first lumbar vertebra. PMID- 10810485 TI - Extensile posterior approach to the radius. AB - An extensile posterior approach to the radius was studied on 20 forearm anatomic specimen upper limbs. The skin incision followed a line from the lateral epicondyle of the humerus to a point corresponding to the middle of the posterior aspect of the wrist. Dissection was done between the extensor digitorum communis and the extensor carpi radialis brevis. The posterior interosseous nerve was identified, and the muscle fibers of the superficial head of the supinator were divided from distal to proximal to the posterior interosseous nerve, which was dissected and carefully retracted laterally. The muscle fibers of the deep head of the supinator were divided to the bone. An incision was made along the superior and inferior margins of the abductor pollicis longus and extensor pollicis brevis. A nerve tape was placed around the two muscles, and they were retracted proximally and medially or distally and laterally, as necessary. To expose the distal third of the radius, the obliquely placed muscles, abductor pollicis longus, and extensor pollicis brevis were retracted proximally and medially. Dissection was done between the extensor carpi radialis brevis and extensor pollicis longus. Anatomic study of the posterior interosseous nerve branches was done to understand the vulnerability of such branches seen in this approach. PMID- 10810486 TI - Radial and axillary nerves. Anatomic considerations for humeral fixation. AB - Because the axillary and radial nerves can be injured during operative exposure and fixation of the humerus, accurate delineation of their location is vital to avoid complications. The authors investigated the relationship of the radial and axillary nerves for radiographically and surgically identifiable bony landmarks. Fifty fresh human cadaveric upper extremities were dissected to identify the nerves as they crossed the lateral intermuscular septum and the humeral surgical neck, respectively. Longitudinal distances between the nerves and the superior aspect of the humeral head, the surgical neck, the superior extent of the olecranon fossa, and the distal aspect of the trochlea were measured with calipers. The average distance from the axillary nerve to the proximal humerus was 6.1 +/- 0.7 cm (range, 4.5-6.9 cm) and 1.7 +/- 0.8 cm (range, 0.7-4.0 cm) from the surgical neck. The radial nerve traversed the lateral intermuscular septum 17 +/- 2.3 cm (range, 13-22 cm) from the proximal humerus, 12 +/- 2.3 cm (range, 7.4-16.6 cm) from the olecranon fossa, and 16 +/- 0.4 cm (range, 9.0-20.5 cm) from the distal humerus, representing the approximate midpoint of the bone. Anteroposterior locking screws placed into the proximal humerus endanger the axillary nerve because it lies directly over the posterior cortex as little as 0.7 cm from the surgical neck. As the radial nerve crosses the lateral intermuscular septum more proximal than generally was thought, it is at risk during implant insertion in the distal half of the humerus. Using measurements calculated from preoperative and intraoperative imaging, the approximate position of the nerve could be determined to better plan fixation method and implant location. PMID- 10810487 TI - The biologic response to laser thermal modification in an in vivo sheep model. AB - The purpose of this study was to evaluate the effect of nonablative laser energy on mechanical, histologic, ultrastructural, and biochemical properties of joint capsular tissue in an in vivo sheep model. Femoropatellar joint capsule was treated with the holmium:yttrium-aluminum-garnet laser via an arthroscope, and tissues were harvested immediately after surgery, or at 3, 7, 14, 30, 60, 90, and 180 days after surgery (n = 8/group). Laser treatment caused significant decreases in tissue stiffness from 0 to 7 days after surgery, then stiffness gradually increased after 14 days. Tissue strength was lowest 3 days after laser treatment. Histologic examination revealed immediate collagen hyalinization and cell necrosis, followed by active cellular response characterized by extensive fibroblast migration and capillary sprouting. Tissue appeared to be normal histologically 60 days after surgery; however, collagen fibrils remained uniformly small. This study showed an active tissue response secondary to thermal modification with concomitant recovery of mechanical properties by 30 days after surgery. Whether the shrinkage or joint stability was maintained with time remains to be evaluated. To clarify the advantages and disadvantages of this technique, a carefully controlled clinical trial with long term followup should be performed. PMID- 10810488 TI - Mechanical properties of small fragment screws. AB - For many years, stainless steel small fragment screws have been produced by one manufacturer. Recently, other implant makers have begun offering similar stainless steel screws. In addition, screw geometry and material composition have been modified in an attempt to produce screws for a wide range of clinical situations. This study compared the mechanical properties of several commonly used small fragment screws. Seven sets of screws were tested mechanically, including three brands of geometrically identical standard stainless steel cortical screws and one brand each of cannulated stainless steel cortical screws, titanium cortical screws, stainless steel cancellous screws, and bioabsorbable polylactic acid screws. Screws from each group were tested for pullout strength, torque to failure, and three-point bending to failure. There were no differences in the mechanical properties of the identical 3.5-mm standard stainless steel cortical screws. No difference in pullout strength was found between the five sets of cortical screws. However, the cancellous screws had 4% to 24% less pullout strength. Torsion tests showed that cannulated stainless steel cortical, titanium cortical screws, stainless steel cancellous screws, and polylactic acid screws failed at significantly less torque than did standard stainless steel cortical screws. Standard stainless steel cortical screws had the highest mean yield point and maximal load at failure of all screws in three-point bending. Other metal screws had lower yield strength and maximal load at failure than did the standard stainless steel cortical screws, and polylactic acid screws had the least bending strength. PMID- 10810489 TI - Analysis of posterior spinal wiring in a validated rabbit model. AB - Mamillary and spinous process base wiring did not improve L5-L6 posterolateral fusion rate, stiffness, or strength in a previously established rabbit model. Specimen radiography significantly underestimated fusion rate when compared with manual palpation (37% versus 68%). Fused specimens were significantly stiffer (67.2 N/mm versus 41.1 N/mm) and stronger (177 N versus 121 N) in tension than were nonfused specimens. Deep wound infection, detected only at the time of sacrifice, apparently was more common with internal fixation (43% versus 21%). Noninfected specimens were significantly stiffer (63.3 N/mm versus 43.0 N/mm) and stronger (176 N versus 107 N) than were infected specimens. PMID- 10810490 TI - Pressure changes during reaming with different parameters and reamer designs. AB - Reaming increases intramedullary pressure and can cause fat intravasation. This study investigated the influence of reaming parameters on pressure changes using the AO, Gray, and Howmedica reamers. Silicone cylindrical shells with 10-, 11-, 16-, 19-, and 20-mm inner diameters were filled with a 1:2 ratio of paraffin and petroleum jelly. The 10- and 11-mm shells were reamed using a 9-mm reamer, the 16 mm shells with a 14-mm reamer, and the 19- and 20-mm shells with an 18-mm reamer. Each experiment was performed seven times while singularly varying driving speeds of 15, 30, and 50 mm per second, with a revolution rate of 150, 250, and 500 revolutions per minute. Using stepwise linear regression analysis, driving speed was the most significant parameter. The lowest pressure increases occurred at the lowest driving speed and the highest revolution rate in all cylinders. The model used for this study did not take into consideration the interaction between the cortical shell and reamer head. The results of this study suggest that reaming should be done at a low driving speed and a high revolution rate using a small cored reamer head, especially during the first reaming steps. PMID- 10810491 TI - The role of journal clubs in orthopaedic surgery residency programs. AB - Journal clubs have a long history in graduate medical education; however, their role in orthopaedic surgery residencies has not been analyzed. The 161 orthopaedic residency program directors in the United States and Canada were surveyed to determine the frequency, format, and goals of journal clubs. Journal clubs are a regular part of the education program in 147 of the 149 programs that responded. Most journal clubs meet once per month in the evening and review articles from more than one periodical. Program directors rate teaching residents how to evaluate a scientific article as the most important goal of a journal club. Although journal clubs are virtually universal in orthopaedic programs, additional study is needed to assess their efficacy and optimal format. PMID- 10810492 TI - Fibular mass in a 28-year-old man. PMID- 10810493 TI - Some unsubstantiated remarks about the United Kingdom training style were made. PMID- 10810494 TI - Melatonin--without the hype. PMID- 10810495 TI - Serotonin N-acetyltransferase. A personal historical perspective. PMID- 10810496 TI - Evolution of melatonin-producing pinealocytes. PMID- 10810497 TI - Melatonin biosynthesis in chicken retina. Regulation of tryptophan hydroxylase and arylalkylamine N-acetyltransferase. PMID- 10810498 TI - Ultraviolet light suppresses melatonin biosynthesis in chick pineal gland. PMID- 10810499 TI - Effects of vasoactive intestinal peptide and histamine on melatonin and cAMP production in chick embryo pineal cells. PMID- 10810500 TI - Ceramide inhibits L-type calcium channel currents in rat pinealocytes. PMID- 10810501 TI - Expression of melatonin receptors and 2-[125I]iodomelatonin binding sites in the pituitary of a teleost fish. AB - The mechanisms of the photoperiodic control of fish physiology (growth, reproduction) and behavior (locomotor activity) are far from being understood. We show here that 2-[125I]iodomelatonin binds specifically to membrane preparations from pike (Esox lucius, L.) pituitaries (KD: 556 pM; Bmax: 2.8 fmol/mg proteins). Radioautography indicated that the binding was restricted to a part of the pituitary only. Using polymerase chain reaction from pike genomic DNA we were able to isolate, subclone and sequence two fragments. The so-called P4 and P8 fragments displayed homology with, respectively, the Mel1a and Mel1b receptor subtypes. The P4 and P8, probes allowed detection of mRNAs corresponding to these receptors in different areas of the brain, including the pituitary. This is the first evidence that melatonin receptors and binding sites are expressed in the pituitary of a non-mammalian species. We suggest that in fish the melatonin mediated photoperiodic control of neuroendocrine functions might involve a direct effect on the pituitary. PMID- 10810502 TI - Melatonin release from the pineals of two sparids. Sparus aurata and Acanthopagrus bifasciatus. PMID- 10810503 TI - Photoendocrine signal transduction in pineal photoreceptors of the trout. Role of cGMP and nitric oxide. AB - This study describes the presence and distribution of cGMP-immunoreactivity and of the nitric oxide (NO) synthesizing enzyme, NO synthase (NOS), as demonstrated by use of the NADPH-diaphorase technique in directly light sensitive pineal organ of the trout. Cyclic GMP immunohistochemistry revealed immunoreactivity in pineal photoreceptor cells that were identified by double-labeling with S-antigen, whereas NADPH-positive structures were located adjacent to these photoreceptor cells. Since NO is known to stimulate synthesis of cGMP, these results indicate a role for NO in pineal function, e.g. in cGMP related events in the phototransduction process as well as in the light-dark control of melatonin synthesis. PMID- 10810504 TI - Intrinsic glutaminergic system negatively regulates melatonin synthesis in mammalian pineal gland. PMID- 10810505 TI - Synaptic vesicle protein SV2B, but not SV2A, is predominantly expressed and associated with microvesicles in rat pinealocytes. PMID- 10810506 TI - Neuropeptide Y (NPY) and NPY receptors in the rat pineal gland. AB - NPY is considered to play an important role in pineal function, because it is co stored with the dominant pineal transmitter noradrenaline. However, little evidence from the literature suggests that NPY alone is a strong regulator of melatonin synthesis or secretion and it is therefore more likely that NPY modulates noradrenergic neurotransmission in the rat pineal gland. The purpose of the present studies was to determine the nature and origin of NPYergic inputs to, and the type of specific NPY receptor subtypes in, the rat pineal gland. Gel filtration and immunocytochemistry using region-specific antisera revealed that all proNPY present in intrapineal nerve fibres is cleaved to amidated NPY and a C terminal flanking peptide of NPY (CPON). The vast majority of NPY content in the pineal gland was found to be of sympathetic origin. Receptor autoradiography showed that only a few NPY specific binding sites were present in the superficial pineal gland. A reverse transcriptase polymerase chain reaction detected sequences of only NPY receptor subtype Y1 and not other NPY receptor subtypes in pineal extracts. These results together with the available literature imply that NPY under certain conditions is co-released with noradrenaline and exerts its actions either presynaptically or on the pinealocyte through a Y1 receptor. The available data indicate that NPY has no effect alone, but acts in concert with noradrenaline. A presynaptic action regulating noradrenaline neurotransmission is also possible. NPY has been reported only to act on melatonin secretion in vitro, and it remains to be established what function NPY plays in the pineal gland in vivo. This paper discuss possible modulatory actions of NPY being a predominant sympathetic transmitter. PMID- 10810507 TI - Signal transduction in the rodent pineal organ. From the membrane to the nucleus. AB - The rodent pineal organ transduces a photoneural input into a hormonal output. This photoneuroendocrine transduction leads to highly elevated levels of the hormone melatonin at night-time which serves as a message for darkness. The melatonin rhythm depends on transcriptional, translational and posttranslational regulation of the arylalkylamine-N-acetyltransferase, the key enzyme of melatonin biosynthesis. These regulatory mechanisms are fundamentally linked to two second messenger systems, namely the cAMP- and the Ca(2+)-signal transduction pathways. Our data gained by molecular biology, immunohistochemistry and single-cell imaging demonstrate a time- and substance-specific activation of these signaling pathways and provide a framework for the understanding of the complex signal transduction cascades in the rodent pineal gland which in concert not only regulate the basic profile but also fine-tune the circadian rhythm in melatonin synthesis. PMID- 10810508 TI - Regulation of melatonin synthesis in the ovine pineal gland. An in vivo and in vitro study. PMID- 10810509 TI - Melatonin modulation of prolactin and gonadotrophin secretion. Systems ancient and modern. AB - Recent studies in sheep indicate that the pineal melatonin signal which transduces effects of photoperiod acts at separate sites in the pituitary gland and brain to regulate seasonality in prolactin (PRL) and gonadotrophin secretion. The pituitary gland is the proposed site for control of PRL based on the observation that hypothalamo-pituitary disconnected (HPD) rams continue to show normal patterns of PRL secretion in response to changes in photoperiod or treatment with melatonin. Lactotrophs do not express melatonin receptors, thus this pituitary effect is assumed to be mediated by cells in the pars tuberalis via "tuberalin". The mediobasal hypothalamus (MBH) is the putative target for gonadotrophin control since: i) gonadotrophin secretion is dependent on pulsatile GnRH secretion from the MBH, ii) local administration of melatonin in the MBH, but not in other areas of the brain and pituitary gland, readily reactivates GnRH induced LH and FSH secretion in photo-inhibited rams; and iii) treatment of HPD rams with a chronic pulsatile infusion of GnRH stimulates gonadotrophin secretion irrespective of photoperiod. Complementary studies conducted by others in the Syrian hamster, have shown that lesions in the MBH block the action of melatonin on gonadotrophin but not on prolactin secretion; this supports the "dual-site hypothesis". Since all photoperiodic mammals are essentially similar in hyper secreting PRL under long days, the pituitary control mechanism for PRL is regarded as conserved (ancient) with the pleiotrophic actions of PRL inducing a summer physiology (e.g. growth of summer pelage). In contrast, the variation between species in the timing of the gonadal cycle indicates that evolution has independently modified the melatonin-sensitive neural circuits in the MBH to permit the species-specific timing of the mating season. PMID- 10810510 TI - The Siberian hamster as a model for study of the mammalian photoperiodic mechanism. AB - The Siberian hamster has been a useful model for studies of mammalian photoperiodism for a number of reasons: 1) Siberian hamsters are hardy animals that are easily maintained and bred in the laboratory. 2) The species exhibits a large number of seasonal, photoperiod-driven, pineal-dependent responses. Thus, the Siberian hamster is an excellent species in which to examine whether several different types of photoperiod responses share similar mechanistic features with respect to their control by MEL. Are all the responses cued to the duration of the nocturnal MEL peak? Does MEL act at a single site to influence all the types of responses, or are there separate MEL target sites for different responses? 3) Juvenile Siberian hamsters exhibit an unusually rapid (for mammals) response to photoperiod change or to MEL treatments, making them ideal subjects for certain types of photoperiod-related studies. 4) Populations of Siberian hamsters show individual variations in photoperiod responsiveness, and the differences are at least partly heritable. These hamsters also exhibit strong influences of environmental history on short day responsiveness. Thus, the species may be a valuable model for the investigation of both genetic and environmental influences on the photoperiodic mechanism. 5) Siberian hamsters have proved to be useful animals in which to study maternal influences on the developing photoperiodic mechanism of the fetus. PMID- 10810511 TI - How does the melatonin receptor decode a photoperiodic signal in the pars tuberalis? PMID- 10810512 TI - Daily and circadian expression patterns of mt1 melatonin receptor mRNA in the rat pars tuberalis. PMID- 10810513 TI - Molecular pharmacology and function of melatonin receptor subtypes. PMID- 10810514 TI - Mechanisms of melatonin action in the pituitary and SCN. AB - We have compared melatonin effects in two different cell types in order to determine general intracellular mechanisms of its action. In neonatal rat pituitary, melatonin acts via the specific membrane receptors to inhibit GnRH induced LH release. The melatonin effect disappears in adulthood due to the disappearance of the receptors. The mechanism of the melatonin action involves inhibition of the GnRH induced increase of intracellular calcium ([Ca2+])i. Our observations indicate that melatonin has dual inhibitory effect on GnRH-induced [Ca2+]i: it inhibits mobilisation of Ca2+ from endoplasmic reticulum as well as Ca2+ influx through voltage sensitive channels. Besides, melatonin also decreases basal and GnRH- or forskolin-induced increase of cAMP concentration in the pituitary. Although cAMP is not of primary importance for regulation of LH release, the cAMP decrease may participate in the mechanism of inhibitory melatonin action on LH release. Rat suprachiasmatic nuclei (SCN) have a high density of the melatonin receptors throughout the postnatal life. Cultures of dispersed SCN cells show circadian rhythm of vasopressin (AVP) release, with several fold increase in the middle of the day and decrease during night. Melatonin inhibits the spontaneous AVP release. Melatonin also inhibits the AVP release induced by vasoactive intestinal peptide (VIP). Intracellular mechanisms of the melatonin effect may involve cAMP, because melatonin inhibits the VIP induced increase of cAMP and increase of cAMP formation by forskolin stimulates AVP release from the cultures. On the other hand, involvement of intracellular calcium in the regulation of AVP release may not be excluded. VIP induces [Ca2+]i increase in 14% of the SCN cells and AVP release is stimulated by Ca2+ ionophore ionomycin. Our observations indicate that some of the mechanisms of melatonin action are similar in the pituitary and SCN. PMID- 10810515 TI - The roles of melatonin in development. PMID- 10810516 TI - Investigation of the human Mel 1a melatonin receptor using anti-receptor antibodies. PMID- 10810517 TI - A pharmacological interaction between melatonin and the alpha 2-adrenoceptor in cuckoo wrasse melanophores. PMID- 10810518 TI - SCN cells expressing mt1 receptor mRNA coexpress AVP mRNA in Syrian and Siberian hamsters. PMID- 10810519 TI - Suprachiasmatic nuclei, intergeniculate leaflet, and photoperiod. PMID- 10810520 TI - Comparison of the pineal and SCN rhythmicity. Effect of photic and non-photic stimuli, photoperiod, and age. PMID- 10810521 TI - Melatonin normalizes the re-entrainment of senescence accelerated mice (SAM) to a new light-dark cycle. PMID- 10810522 TI - Developmental expression of both melatonin receptor mt1 mRNA and melatonin binding sites in Syrian hamster suprachiasmatic nuclei. PMID- 10810523 TI - Entrainment of rat circadian rhythms by daily administration of melatonin. Influence of the mode of administration. PMID- 10810524 TI - Melatonin as a chronobiotic for circadian insomnia. Clinical observations and animal models. PMID- 10810525 TI - Melatonin and cardiovascular function. PMID- 10810526 TI - The effect of melatonin on vasopressin release under stress conditions in pinealectomized male rats. AB - The findings here reported showed that the response of vasopressinergic neurons to immobilization stress is augmented by melatonin. The effectiveness of melatonin in functional modification of these neurons' activity under conditions of stress changes after pineal removal. PMID- 10810527 TI - Motility and passive avoidance modulation by septal vasopressin is dependent on the pineal gland. PMID- 10810528 TI - Effects of melatonin and its relation to the hypothalamic-hypophyseal-gonadal axis. PMID- 10810529 TI - Melatonin influence upon ovary during ageing. A morphometric study. PMID- 10810530 TI - Melatonin effect during aging on reproductive hormones of female rats through the estrous cycle. PMID- 10810531 TI - New insights into melatonin regulation of cancer growth. PMID- 10810532 TI - Melatonin synergizes with retinoic acid in the prevention and regression of breast cancer. PMID- 10810533 TI - Melatonin and 9-cis-retinoic acid in the chemoprevention of NMU-induced rat mammary carcinoma. PMID- 10810534 TI - The antiproliferative effects of melatonin on experimental pituitary and colonic tumors. Possible involvement of the putative nuclear binding site? PMID- 10810535 TI - Cytochalasin B influence on megakaryocyte patch-clamp. PMID- 10810536 TI - Relationships between melatonin, glutathione peroxidase, glutathione reductase, and catalase. Endogenous rhythms on cerebral cortex in Gallus domesticus. AB - In this report, we studied the endogenous rhythms of three antioxidant enzymes: glutathione peroxidase (E.C.1.11.1.9), glutathione reductase (E.C.1.6.4.2) and catalase (E.C.1.11.1.6) in cortex of chick brain and correlate them with physiological blood melatonin concentrations. PMID- 10810537 TI - Effect of pinealectomy on melatonin levels in the gastrointestinal tract of birds. PMID- 10810538 TI - Melatonin and 5-methoxytryptamine in the bioluminescent dinoflagellate Gonyaulax polyedra. Restoration of the circadian glow peak after suppression of indoleamine biosynthesis or oxidative stress. PMID- 10810539 TI - Presence and possible role of melatonin in a short-day flowering plant, Chenopodium rubrum. PMID- 10810540 TI - MLT and the immune-hematopoietic system. AB - It is now well recognized that a main actor in the continuous interaction between the nervous and immune systems is the pineal hormone MLT. T-helper cells bear G protein coupled MLT cell membrane receptors and, perhaps, MLT nuclear receptors. Activation of MLT receptors enhances the release of T-helper cell type 1 (Th1) cytokines, such as gamma-interferon and interleukin-2 (IL-2), as well as of novel opioid cytokines which crossreact immunologically with both interleukin-4 (IL-4) and dynorphin B. MLT has been reported also to enhance the production of interleukin-1 (IL-1), interleukin-6 (IL-6) and interleukin-12 (IL-12) in human monocytes. These mediators may counteract stress-induced immunodepression and other secondary immunodeficiencies, protect mice against lethal viral and bacterial diseases, synergize with IL-2 in cancer patients and influence hematopoiesis. In cancer patients, MLT seems to be required for the effectiveness of low dose IL-2 in those neoplasias that are generally resistant to IL-2 alone. Hematopoiesis is apparently influenced by the action of the MLT-induced-opioids (MIO) on kappa-opioid receptors present on stromal bone marrow macrophages. Most interestingly, gamma-interferon and colony stimulating factors (CSFs) may modulate the production of MLT in the pineal gland. A hypothetical pineal-immune hematopoietic network is, therefore, taking shape. From the immunopharmacological and ethical point of view, clinical studies on the effect of MLT in combination with IL-2 or other cytokines in viral disease including human immunodeficiency virus-infected patients and cancer patients are needed. In conclusion, MLT seems to play a crucial role in the homeostatic interactions between the brain and the immune-hematopoietic system and deserves to be further studied to identify its therapeutic indications and its adverse effects. PMID- 10810541 TI - The use of melatonin and co-treatment with autologous or allogeneic cells as a model for control of malignant beta-cell leukemia. PMID- 10810542 TI - Autoradiographic detection of 2-(125I)-iodomelatonin binding sites in immune tissue of rats. PMID- 10810543 TI - Is melatonin a photoperiodic signal in humans? PMID- 10810545 TI - Melatonin and aging. PMID- 10810544 TI - Melatonin as a marker and phase-resetter of circadian rhythms in humans. PMID- 10810546 TI - Phase of melatonin rhythm in winter depression. PMID- 10810547 TI - Influence of low-frequency magnetic field of different characteristics on serum melatonin concentrations in humans. PMID- 10810548 TI - Circadian serum melatonin profiles in patients with very large goitre before and after surgery. Preliminary report. PMID- 10810549 TI - Advanced immunoassays for the direct determination of melatonin in human serum and culture media. PMID- 10810550 TI - Review article limb salvage surgery in bone tumours. AB - Wide resection of bone tumour has become an accepted treatment in the limb salvage surgery. The reconstruction of the residual defect following wide resection is a major problem. The motions of involved joint after resection of the large tumour can be restored by using either the osteoarticular allograft or endoprosthesis. Osteoarticular graft is suitable for proximal tibial reconstruction and endoprosthesis for distal femoral reconstruction. Resection arthrodesis can be done with autograft, cement or autoclaved tumour bone. Autograft is rarely used to reconstruct the large residual defect. Cement can reconstruct the larger defect, but it is not a suitable procedure on long term basis. The reimplantation of resected autoclaved tumour bone graft is technically a simple and financially a cost saving solution for this difficult problem. It is the most suitable method of reconstruction for the developing and poor countries where the resources for other methods are not available due to financial technical or socio-cultural reasons. PMID- 10810551 TI - Borderline epithelial tumours of the ovary--a retrospective analysis of 31 cases. AB - Thirty one cases of epithelial borderline tumours of the ovary recorded over a period of six years were reviewed. The incidence of borderline tumours was 6% in relation to ovarian epithelial malignancies, with serous and mucinous types comprising three fourth of the lesions. The serous tumours were bilateral in 39%, revealed surface growth in 17% and had peritoneal implants in 11% of cases. The mucinous tumours were bilateral in 11% and had associated pseudomyxoma peritonei in 22% of cases. Nuclear grade appeared to correlate with extraovarian spread and surface growth in the serous borderline tumours, but not in the mucinous borderline tumours. The endometrioid borderline tumours and mixed epithelial borderline tumours were rare lesions. Twenty one patients (68%) presented in Stage-la. Surface growth correlated with recurrences. The prognosis remained good in serous borderline tumours even in the presence of implants as these were non invasive. The mean disease free survival was 43.03 months. There was no statistical difference in disease free survival of patients with and without implants. PMID- 10810552 TI - Granulocyte macrophage--colony stimulating factor (GM-CSF) as adjunct in induction therapy of acute myeloid leukemia. AB - Use of growth factors (G-CSF/GM-CSF) as adjunct in induction therapy of AML is controversial. Possible stimulation of leukemia cell clones has been the major cause of concern. We treated 50 cases of AML with GM-CSF as an adjunct during induction therapy. 35 patients (70%) achieved complete remission out of which 13 patients relapsed at a median relapse period of 15 months. Average duration of neutropenia was 10.5 days. (15 days in the control) Febrile episodes were fewer and antibiotic support was required for an average period of only 7.6 days (16.9 days in the control). The benefits including the economic analysis of the role of GM-CSF in this setting is discussed. PMID- 10810553 TI - Cryptosporidium and isospora belli diarrhoea in immunocompromised hosts. AB - Cryptosporidium and isospora, two of the intestinal coccidian parasites known to be the causative agents of acute diarrhoea in animals, have now emerged as one of the main causes of prolonged life threatening diarrhoea in immunocompromised patients particularly so in patients with AIDS. Between June 1996 and December 1997, a total of 75 immunocompromised patients presenting with diarrhoea were investigated both for Cryptosporidium and Isospora. The study group consisted of cancer and AIDS patients with history of diarrhoea. Cryptosporidium oocysts were detected in 35 patients (46.7%). One of the faecal samples from an AIDS patient with diarrhoea showed the presence of both Cryptosporidium and Isospora oocysts. To the best of our knowledge, this is the second documented report of Isospora associated diarrhoea in an AIDS patient from India. The various techniques used for demonstration of these parasites were modified acid fast staining, Safranine Methylene-blue staining and direct immunofluorescence test. PMID- 10810554 TI - Role of high speed biopsy gun in breast cancer diagnosis. AB - Preoperative diagnosis of histology and receptor status is important in management of breast cancer. Percutaneous automated core biopsy with biopsy gun (Gun biopsy) was done in fifty patients with palpable breast lump in whom fine needle aspiration cytology (FNAC) was either negative or not done In all patients adequate tissue for histology and receptor status studies was obtained forty-two patients had infiltrating duct carcinoma and eight patients had benign lesions on gun biopsy. There were no complications in this procedure. Twenty of the forty two patients underwent mastectomy either per primum or after chemotherapy, had the diagnosis substantiated on histopathological examination Thirty-four samples were examined for receptor status and the specimen was found to be adequate and of good quality. We conclude that gun biopsy is a simple and safe procedure which is more sensitive and specific than FNAC. PMID- 10810555 TI - Adjuvant therapy in invasive thymoma: an audit of cases treated over an 8 year period. AB - Invasive thymomas comprise 0.1%-0.2% of all malignancies in India. This report is an audit of 11 cases (10 males and 1 female) at a mean age of 36.6 years (range 25-52 years) of invasive thymoma accrued over an eight year period treated by combined modality treatment. Nine of these presented with myaesthenia gravis. All patients underwent initial surgery (3 partial and 8 total resections) and postoperative radiotherapy. Two of the three partially resected patients received one course of chemotherapy prior to radiotherapy consisting of cyclophosphamide, vincristine, procarbazine and prednisolone. At a median follow up of 28 months (range 2-87) there have been no local relapses, one distant metastasis and one death due to uncontrolled myaesthenia. The treatment strategies with invasive thymomas would depend upon the extent of resection. Postoperative radiotherapy appears to be indicated in all cases, however the role of chemotherapy may be limited to those with partial resection. PMID- 10810556 TI - Value of endometrial aspiration cytology in assessing endometrial status in symptomatic peri and postmenopausal women. AB - Endometrial aspiration cytology and curettage were done on fifty one patients, randomly selected from three hundred and nineteen peri and post menopausal patients, who presented with menstrual cycle related complaints. The material obtained on aspiration was made into smears and cell block. The diagnoses of the aspirate and the curettage were made independently and later correlated The sensitivity of the procedure was 81.63% and the specificity was 83.33% A significant correlation was observed between cytologic and histopathologic diagnosis. In hyperplastic states, while on cytology, the diagnosis was made, the degree of hyperplasia was possible only in cell block and in paraffin sections. Thus the low cost feasibility and the sensitivity makes endometrial aspiration a useful diagnostic procedure for screening the endometrial status. However, a negative result does not rule out a malignant process, especially in persistently symptomatic patients and further investigations are mandatory. PMID- 10810557 TI - Effects of substance P on nicotine-induced intracellular Ca2+ dynamics in bovine adrenal chromaffin cells. AB - Substance P (SP) is colocalized with ACh in splanchnic nerves that innervate into adrenal medulla and the peptide has been shown to inhibit nicotinic agonists induced catecholamine secretion. To elucidate the effects of SP on cytosolic Ca2+ dynamics, the present study was conducted using fura-2-loaded isolated bovine adrenal chromaffin cells. Stimulation of the cells with nicotine (10-100 microM) produced a rapid rise of cytosolic Ca2+ concentration ([Ca2+]i), the peak level of which increased in a dose-dependent manner, followed by a gradual decay. In the presence of 10 microM SP, the dose-response relationship of the peak levels shifted downward. Quantitative analyses implied that SP inhibits the nicotine induced Ca2+ influx in a noncompetitive manner. Nicotinic acetylcholine receptor is composed of two major functional domains: an agonist-binding site and an ionophore or channel domain. Agonist binding activates ionophore/channel domain and causes mainly Na+ influx. This Na+ influx depolarizes the cell and activates voltage-dependent Ca2+ channels. Based on this fact, the present results indicate that SP dose not block nicotine binding sites but interferes with other sites of nicotinic receptor/channel molecule, most probably a channel domain. It was suggested that SP colocalized with ACh in splanchnic nerves functions as a physiological modulator of catecholamine secretion by non-competitively suppressing ACh-induced cytosolic Ca2+ dynamics in bovine adrenal chromaffin cells. PMID- 10810558 TI - Effects of concurrent exposure to 3-methylcholanthrene and vitamin A on fetal development in rats. AB - To investigate the effect of the environmental pollutants, polycyclic aromatic hydrocarbons (PAHs), on retinoic acid-induced teratogenesis, all-trans-retinoic acid (RA) dissolved in corn oil (120 mg/kg) was administered orally to pregnant rats at the 11th day of gestation with and without the prior intraperitoneal treatment with 10 mg/kg 3-methylcholanthrene (3-MC) for 3 days. Dams were killed on the 20th day of pregnancy. The examinations of fetuses revealed that 3-MC barely enough to cause induction of P-450 in pregnant dams had profound embryo toxic effects: the fetal resorption amounted to approximately 60% of total number of implantations. The fetuses survived weighed less than the control fetuses. All of RA-treated mothers had fetuses with abnormalities, and the main malformations were absence of tail (100%), caudal and sacral malformations (100%), and cleft palate (42%). Pregnant dams received both 3-MC and RA had a reduced severity of tail anomaly (33%), while the rest, 67%, had short vestigial tail. Caudal and sacral malformations were detected but at a milder degree. We did not observe cleft palate in this group. The concurrent treatment of dams with 3-MC and RA led to an increased inducibility of cytochrome P-450 and subsequently, CYP1A1 dependent enzyme activity higher than those observed after the injection of 3-MC alone. UDP-glucuronyl-transferase activity was also markedly induced in concurrent 3-MC and RA group higher than that in 3-MC alone. We suggest that the induction of P-450 and alteration of metabolic enzyme activities may play an important role in reducing the teratogenic potency of RA. However, RA-treatment did not retard the embryo-toxic effect of 3-MC but rather potentiated. PMID- 10810559 TI - Protective effects of intranasal vaccination with plasmid encoding pseudorabies virus glycoprotein B in mice. AB - Intranasal administration of plasmid DNA encoding glycoprotein B of pseudorabies virus into mice induced both serum and secretory antibody responses. These mice resisted intranasal challenge with lethal dose of the virus, but did not intraperitoneal challenge. On the other hand, intramuscular injection of the plasmid induced less secretory and higher serum antibody responses than those of intranasally vaccinated mice. None of them was protected from virus challenge. The present results suggest that administration of plasmid DNA encoding glycoprotein B by respiratory mucosal route generates local secretory antibodies which serve to protect animals from pseudorabies virus infection. PMID- 10810560 TI - Epidemiological importance of younger age group during malaria epidemic in PHC Tamulpur, Assam. AB - An investigation of a malaria epidemic was carried out in Tamulpur Primary Health Centre, district Nalbari, Assam during April 1995. The analysis revealed that children between 3 and 12 years of age who were treated and who recovered clinically from fever during the epidemic were instrumental in the progression of the epidemic by acting as Plasmodium falciparum gametocyte reservoirs. Special efforts are required for treatment of children below 12 years during an epidemic. PMID- 10810561 TI - Bronchopulmonary infection due to Moraxella (Branhamella) catarrhalis at a specialist hospital in Saudi Arabia. AB - Moraxella (Branhamella) catarrhalis is now considered as one of the most important causative organisms responsible for respiratory tract infection. Specimens of tracheal aspirates from inpatients at King Fahd specialist hospital, Buraidah, Saudi Arabia were collected over a period of 18 months to determine prospectively the frequency of Moraxella catarrhalis and its antimicrobial susceptibility. Moraxella catarrhalis was isolated in pure culture from 3.8% of the tracheal aspirates collected from patients with symptoms of acute respiratory tract infections. It was the third most important pathogen after Haemophilus influenzae and Streptococcus pneumoniae; 76% of the patients had an underlying chronic obstructive pulmonary disease (COPD). The significance of the findings is discussed. PMID- 10810562 TI - The prevalence of Toxoplasma gondii specific IgG and IgM antibodies in blood donors in Al Ain, United Arab Emirates indicates a potential risk to recipients. AB - The results of a prospective cross-sectional study on the anti-toxoplasma IgG and IgM specific antibody profile among blood donors in Al Ain United Arab Emirates are presented. The overall infection rate was 34%. Based on IgM specific antibody positive rate, acute toxoplasmosis was evident among 3% of the blood donors studied. The theoretical implications of these findings are discussed. PMID- 10810563 TI - Acid fast staining versus ELISA for detection of Cryptosporidium in stool. AB - A prospective study was undertaken to compare the efficacy of ELISA in detection of Cryptosporidium Specific Antigens (CSA) in stool specimens of patients attending various OPDs in a Delhi Hospital. A total of 216 consecutive faecal specimens were examined microscopically after modified acid fast staining. An ELISA was also performed using ELI-WFLL CRYPTO detection kit following the manufacturer's instructions for detection of CSA in stool specimens. Taking microscopy as the gold standard ELISA was found to be 100% sensitive and 99.07% specific in detection of Cryptosporidium spp. The test is easy to perform and interpret. PMID- 10810564 TI - Re-infection estimation of soil-transmitted helminths among slum school children in Visakhapatnam, Andhra Pradesh. AB - Prevalence and intensity study of soil-transmitted helminths was carried out in a primary municipal corporation school in Relliveedhi, Visakhapatnam, Andhra Pradesh. All subjects (n = 217) belonged to low socio-economic status with an age distribution of 7-13 years. Reinfection study was estimated over a nine month period following treatment with albendazole administered at a single oral dose of 400 mg/child. Parasites identified were Ascaris lumbricoides, Trichuris trichiura and hookworm. Post-treatment prevalence reached pre-intervention level but the intensity of infection remained appreciably low (p < 0.05). PMID- 10810565 TI - Maternal knowledge on risk behavioural practices and it's association with diarrhoea in a rural community of West Bengal, India. AB - Knowledge of rural mothers related to five diarrhoeagenic risk behaviours, identified in an earlier study, was ascertained. A high proportion of mothers (67%-79%) had knowledge about risk of bottle feeding, non-use of soap for cleaning feeding containers, storage of drinking water in wide-mouthed vessels and indiscriminate disposal of children's faeces. However, only around 31% of mothers were aware about danger of using pond water for cleaning feeding containers. Risk behavioural practices were less amongst mothers who had knowledge about them. Risk of diarrhoea amongst children of mothers having risk practice without knowledge as compared to those who utilised their knowledge to avoid risk practice was found significantly higher (p < or = 0.005) except for bottle feeding (p = 0.330). The results of this study indicate that children can be protected significantly from diarrhoea if mothers' diarrhoeagenic behaviours can be altered through educational intervention. PMID- 10810566 TI - Merit of mobilization technique in transfer of non-autotransferable R-plasmids. AB - A total of 200 urinary isolates of Esch.coli received at National Salmonella and Escherichia Centre, Central Research Institute, Kasauli during the years 1995 to 1997 were studied for transferable drug resistance. Out of 188 strains, 134 strains showing resistance to either Nalidixic acid or Rifampicin were subjected to autotransferable resistance studies. Of these 134 strains 131 showed either partial or enbloc transfer of R-factor. Mobilization experiment successfully transferred resistance marker in 14 of the 68 isolates in which resistance to one or more drugs could not be transferred during conjugation experiment. PMID- 10810567 TI - Colicinogenicity and enteropathogenicity of Escherichia coli associated with gastroenteritis in man. AB - Sixty strains of Escherichia coli, were isolated and identified from 94 samples of infantile diarrhoea and traveller's diarrhoea. Eleven strains were found to be colicinogenic and colicin 'H' was found to be predominant in conjunction with other colicins. Five strains were enteropathogenic of which 055 and one untypable strain were highly enteropathogenic. A positive correlation could be established between haemolytic character, dulcitol fermentation and the virulence of the strain involved. PMID- 10810568 TI - Some aspects of changing behaviour of malaria vectors in tribal areas of India. AB - The field entomological studies and surveys carried out in 72 tribal districts out of 100 in seven penninsular States namely, Andhra Pradesh, Bihar, Gujarat, Madhya Pradesh, Maharashtra, Orissa and Rajasthan indicated that in most of the areas the vector mosquitoes encountered during the present study were almost the same as recorded by the earlier workers except that Anopheles fluviatilis James, 1902 was recorded in the areas of district Raipur, Durg, Bilaspur, Raigarh, Shahdol, Rajnandgaon, Barwani and Janjgir of Madhya Pradesh, and Sundergarh of Orissa during the present survey. This vector was not recorded in earlier studies by other workers. An. culicifacies Giles, 1901 was not found in present survey in East Godavari and Vishakhapatnam of Andhra Pradesh and Ganjam and Koraput of Orissa where this mosquito was found in collections earlier. Similarly, An. fluviatilis was not encountered during the present field study in Bhilwara of Rajasthan, Phulbani and Kalahandi of Orissa, Thane, Nanded and Nasik of Maharashtra and Bharuch of Gujarat state. During this study An. fluviatilis was noted to be mostly endophilic whereas earlier workers noted this mosquito to be exophilic in a large number of districts. The majority of the tribal districts seem to be under the influence of two malaria vectors, An. culicifacies and An. fluviatilis and these tribal districts are maintaining a high malaria endemicity with predominance of Plasmodium falciparum infection. In the present study, the transmission seasons were noted to be longer than recorded earlier in the districts of Andhra Pradesh, Gujarat, Rajasthan and Orissa. In the remaining states it is almost similar as recorded earlier. PMID- 10810569 TI - Prevalance and resistance pattern of Salmonella serotypes in India. AB - During the period 1990-91, 3222 Salmonella strains were identified at the National Salmonella and Escherichia Centre (NSEC) at Central Research Institute, Kasauli. Of these, 2894 were from humans, 226 from poultry, 84 from animals and remaining 18 from reptiles, birds and other sources. These strains belonged to 53 different serotypes. These include 4 serotypes reported for the first time in India, namely S. kedogou, S. VP. bornheim, S. kisarawe and S. madras. Drug resistance studies revealed that 573 strains were sensitive to all the antibiotics commonly used, 1351 single drug resistant, 594 resistant to two drugs and 704 were multidrug resistant. One strain from human stool was resistant to all the antibiotics used. Prevalence of various Salmonella serotypes and their response to various drugs is discussed. PMID- 10810570 TI - Blood meal identification by using Microdot ELISA in vector mosquitoes. AB - The blood meals of five mosquito species which are vectors for malaria, J.E. and Filaria were determined by using Microdot ELISA technique, which is a rapid and sensitive method. A total of 4354 freshly fed mosquitoes comprising of Anopheles culicifacies, An stephensi, Culex quinquefasciatus, Cx. vishnui gp. and Cx. tritaeniorhynchus were tested. Results of the study reveal that highest proportion of An. culicifacies between 76-100% fed on bovine host followed by 0.3 to 24.2% on human. Cx. vishnui showed 1.9% feeding on human, 49-87.7% on bovine and 7.4-12.3% on the pigs. It has been observed that malaria vectors namely An. culicifacies and An. stephensi are still zoophilic in nature. Similarly, the vector of J.E. namely Culex vishnui gp. and Cx. tritaeniorhynchus also fed on cattle but some samples have indicated feeding of blood meal from pigs. PMID- 10810571 TI - Bacterial agents of acute follicular tonsillitis in Gurayat, Saudi Arabia. PMID- 10810572 TI - Insecticide susceptibility of Anopheles dirus in Assam. PMID- 10810573 TI - Microbiological assay versus spectrophotometry for determination of rifampicin in serum and cerebrospinal fluid. PMID- 10810574 TI - Intestinal parasitic infections in Dharan (Nepal): a hospital based study. PMID- 10810575 TI - Serology for malaria diagnosis in children. PMID- 10810576 TI - Larval ecology of malaria vectors in paddy fields of district Nainital, U.P. PMID- 10810577 TI - Minutes of the Meeting of the General Body of the Indian Society for Malaria and other Communicable Diseases, 22-Shamnath Marg, Delhi 110 054. PMID- 10810578 TI - Central venous catheter related infections. AB - Infusion therapy carries a substantial risk of producing iatrogenic sepsis, bacteremia or fungemia originating from the device used for vascular access or from the contaminated infusate. This study, a semiquantitative assay of the central venous catheter tips (CVC) was carried out and correlated with the blood culture results to determine the probable cause of sepsis in our post operated patients. One thousand six hundred and thirty one surgeries were performed in the Institute of Cardio-Vascular Diseases from January to December 1997. We received 150 CVC tips which were in place for more than five days for culture. Fifty one (51) CVC tips showed growth in either contents or roll. There was associated blood stream infection in 23 patients. The age groups of patients ranged from two months to 67 years and male to female ratio was 109:41. Children below the age of 12 years were 62 in number. Candida spp. (32.4%) and Staphylococcus spp. (33.9%) formed the predominant isolates from the CVC tips. PMID- 10810579 TI - Screening for Chlamydial infection in women attending STD clinic by polymerase chain reaction, ELISA and cell cytology. AB - Thirty five female patients with endocervicitis attending STD clinic were studied for the presence of Chlamydial infection by Polymerase Chain Reaction (PCR), Enzyme Immunoassay (EIA) and Cell Cytology. PCR was found to be positive in 54.2% of patients, Enzyme-Linked Immunosorbent Assay (ELISA) in 25.7% of patients, but cell cytology revealed the presence of inclusion bodies only in 3% of the cases, thereby showing that polymerase chain reaction is a better method for detection of Chlamydia trachomatis than EIA and cell cytology. PMID- 10810580 TI - Immune responses to chloroquine--sensitive and resistant populations of Plasmodium berghei in mice. AB - In order to elucidate the role of the host as a factor in the spread of chloroquine resistance, a study of the host's immune responses in chloroquine resistant (cqr) and chloroquine sensitive (cqs) Plasmodial infections is essential. Course of the infection and the nature of immune responses in mice infected with chloroquine resistant (R) and chloroquine sensitive (S) strains of Plasmodium berghei were compared. Crude parasite antigen activated T cells from both the groups of mice (R and S) and the parasite specific antibodies were detected in the sera of both the groups. The differences in immune responses between the control (uninfected) and infected mice were found to be significant. However, humoral and in vitro cellular responses obtained with T cells from chloroquine resistant P. berghei primed mice was lower in comparison with the responses obtained with T cells from the sensitive infections. Our studies therefore suggest that immunosuppression to parasite antigen is seen in mice primed with chloroquine resistant P. berghei, which may play a role in the development of resistance. PMID- 10810581 TI - Quantitative buffy coat: a special adjunct for diagnosis of malaria. AB - Quantitative buffy coats (QBC) technique was compared with conventional blood film technique for the diagnosis of malaria in a tertiary care hospital. The QBC technique was found to be a rapid technique with a sensitivity of 83% and specificity of 94%. Malaria species identification was also possible. It was essentially very useful to detect parasites below < or = 100 parasites/ul of blood by QBC technique. However, quantification of parasitaemia could not be made using this technique. Many cases of carriers having very few gametocytes in their blood were also identified. It is therefore, concluded that the QBC technique, may be appropriate for screening populations for malaria and for detection of asymptomatic carriers to control further transmission of the disease in the community. PMID- 10810582 TI - The prevalence of HIV in female sex workers in Manipur, India. AB - Data on STDs and sexual practices in commercial sex workers (CSWs) is in general limited in India. Manipur in north-east Indian has a high prevalence of HIV in injecting drug users but the rate in CSWs is not known. The site selected for the study was Moreh, on the Myanmar border of Manipur. One hundred blood samples were collected, 7 from migrants from Myanmar, the remainder from Manipuri women. The HIV seropositivity rate was 12% (95% CI = 5.6-18.4). The age of the women ranged from 15 to 42 (mean = 24.5 years, median 23.7 years). The proportion of HIV positives increased significantly with number of customers per day and number of years in the profession. The HIV prevalence among Injecting drug using CSWs was 9.4 times higher that among non-IDU CSWs. Vaginal discharge was strongly associated with HIV positivity. Effective intervention programmes among CSWs in Manipur to prevent further spread of HIV are strongly indicated by the results of this study. PMID- 10810583 TI - Typhoid fever caused by a typical variants of Salmonella typhi--a preliminary report. AB - In the study carried out from, February, 1996 to January, 1999. 42 (33.9%) a typical variants of S typhi which fermented sucrose were encountered. This variant was identified as of antigenic structure 9, 12, v1, d1 and typed as phage type E1 and Bio type I. PMID- 10810584 TI - Surgical filariasis: immunoscreening for filarial IgG antibodies using Wuchereria bancrofti microfilarial excretory-secretory antigen. AB - A clinical study and immunoscreening was conducted on 363 suspected filarial patients attending the surgery out patient division of the MGIMS, Sevegram. The disease was significantly higher in males (86%) than in females (14%). Majority (52.9%) of the cases were in the age group of 11-30 years. The distribution of cases into three different grades of infection showed, 52.6%, 33.3% and 14.1% of the cases having acute (grade I), sub-acute (grade II) and chronic (grade III) stages of infection respectively. While 73% of the cases had genital manifestations, 23% were with lymphatic obstruction in limbs and the rest of the 4% suffered from manifestations like cellulitis, abscesses, haematuria and chyluria. Filarial IgG antibodies against microfilarial excretory-secretory (mf ES) antigen were detected in 89% of cases with genital manifestations, 87% of lymphoedema cases, 67% of lymphadenitis cases and 60% of cases with other clinical manifestations and 3% of endemic normals. PMID- 10810585 TI - Investigation of an epidemic of acute haemorrhagic conjunctivitis in Pune, India. AB - An epidemic of acute haemorrhagic conjunctivitis (AHC) was reported from Pune in August-September, 1996. Conjunctival swabs and scrapings were taken from acute patients attending the OPD of the Ophthalmology department of B. J. Medical College, Pune. Conjunctival swabs were processed in human embryonic lung fibroblasts (HEL) cell line and viral isolates were obtained which were identified as Enterovirus (EV) 70 prime strain using standard in-vitro neutralisation test. Indirect intracellular immunofluorescence test also confirmed the identity of the virus involved in the epidemic. PMID- 10810586 TI - A high incidence of ciprofloxacin resistance in urinary isolates in eastern Nepal. AB - A study was conducted to examine the incidence of ciprofloxacin resistance in the strains of bacteria isolated from patients with suspected urinary tract infection (UTI) and to analyse the level of resistance. A total of 63 bacterial isolates were grown from quantitative urine culture and were in significant count. Antibiotic susceptibility tests were done by using disc diffusion method of Kirby Bauer and the level of resistance was analyzed by break point minimum inhibitory concentration (MIC). Majority of bacteria isolated and tested were Gram negative with Escherichia coli as the commonest isolate. Antibiotic susceptibility testing revealed a high prevalence of resistance to nalidixic acid, ampicillin and norfloxacin. PMID- 10810588 TI - Tolerability of a recombinant DNA hepatitis B vaccine--results of post-marketing surveillance. PMID- 10810587 TI - An El Tor cholera outbreak in Maldah district, West Bengal. AB - An outbreak of cholera occurred in Maldah district, West Bengal during July August 1998. Attack rate was 34/1000. Cases were more (59.3%) amongst adults (> 15 years.). V. cholerae 01 biotype E1 Tor serotype ogawa was isolated as a single pathogen from 52.9% (9/17 samples examined). All V. cholerae strains belonged to phage type 2 (Basu and Mukherjee scheme) and type 27 (new phage type scheme). The strains were resistant to co-trimoxazole, furazolidone, ampicillin, streptomycin and nalidixic acid. PMID- 10810589 TI - Emergence of multiresistant bacterial pathogens--isolated from endotracheal aspirates. PMID- 10810590 TI - Bancroftian filariasis in a weaving community of lower Assam. PMID- 10810591 TI - An outbreak of malaria in district Maharajganj--an outcome of neglected surveillance. PMID- 10810592 TI - Some emerging food and water borne pathogens. AB - Emerging pathogens are those infective organisms whose incidence has recently increased or is likely to increase during the next two decades due to changes in demography, food habits, food technology, commerce, water sources and environmental factors. Some important emerging food and water borne bacterial pathogens include Listeria monocytogenes, Campylobacter jejuni, Yersinia enterocolitica, Salmonella enteritidis, Escherichia coli O157: H7, Vibrio cholerae biotype E1 Tor Serotype 0139, Vibrio parahaemolyticus and Aeromonas hydrophila, A. sobria, and A. caviae. The prevalence, ecological relationships of these organisms, their transmission through food, water and other environmental sources, and role of their virulent factors in the pathogenesis of infections and their public health significance is discussed in this paper with special reference to the situation in India. PMID- 10810593 TI - A new focus of visceral leishmaniasis in sub-Himalayan (Kumaon) region of northern India. AB - A localised focus of visceral leishmaniasis is reported for the first time from Kumaon region of North India. During a period of 18 months from July 1997 to December 1998 several cases of pyrexia of unknown origin with unresponsiveness to antibiotics and anti-tubercular treatments were referred to the All India Institute of Medical Sciences, New Delhi. Five of these were diagnosed to have visceral leishmaniasis. One of these was also co-infected with HIV-1. All the five cases came from an adjoining area falling under Almora (4) and Nainital(1) districts. Of these four were males and one female. One patient died within 48 hours of hospitalization at AIIMS while rest of four responded completely to sodium stibogluconate therapy. PMID- 10810594 TI - 'In vitro' antiviral activity of neem (Azadirachta indica. A. Juss) leaf extract against group B coxsackieviruses. AB - The antiviral and virucidal effect of methanolic extract fraction of leaves of neem (Azadirachta indica A. Juss) (NCL-11) was studied regarding its activity and possible mechanism of action against Coxsackie B group of viruses. NCL-11 inhibited plaque formation in 6 antigenic types of Coxsackie virus B at a concentration of 1000 micrograms/ml at 96 hrs. 'in vitro'. Additionally virus inactivation, yield reduction and effect of time of addition assays suggested that NCL-11 was most effective against coxsackie virus B-4 as a virucidal agent besides interfering at an early event of its replicative cycle. The evidence suggested that presence of a battery of compounds besides flavonoids, triterpenoids and their glycosides in NCL-11 have antiviral action for coxsackie B group of viruses 'in vitro.' The minimal inhibitory concentrations were not toxic to Vero (African green monkey kidney), cells; subtoxic concentration was 8,000 micrograms/ml and cytotoxic concentration 10,000 micrograms/ml, which was confirmed by trypan blue dye exclusion test. PMID- 10810595 TI - Evaluation of cyfluthrin and fenfluthrin for their insecticidal activity against three vector mosquitoes. AB - The EC50/EC90 concentrations of cyfluthrin and fenfluthrin were tested for their activity against different developmental stages of three important vector mosquitoes viz., Anopheles stephensi Liston, Aedes aegypti (Linn.) and Culex quinquefasciatus Say. The EC90 concentrations of both cyfluthrin and fenfluthrin showed ovicidal effect on An. Stephensi and Ae. aegypti whereas EC90 of cyfluthrin checked the hatching of eggs completely in Cx. quinquefasciatus. Fenfluthrin at EC50 concentration reduced the percentage of hatching significantly (p < 0.05) only in An. stephensi. Both the compounds were more active against the fourth larval instars of all mosquito species and cyfluthrin in culicines (17.3% Ae. aegypti = 9.1%) and fenfluthrin in anophenlines (An. stephensi = 36.8%) brought about maximum inhibition in adult emergence. Various types/degrees of morphogenetic aberrations were induced in all mosquito species on treatment with these compounds. Cyfluthrin treated female mosquitoes showed reduced fecundity rates in An. stephensi (p < 0.05), Cx. quinquefasciatus (p < 0.001) and fenfluthrin treated in An. stephensi (p < 0.5) and Ae. aegypti (p < 0.05). The fertility rates of all the mosquito species were significantly reduced (p < 0.001) by both the compounds. PMID- 10810596 TI - Lymphatic filariasis in two distinct communities of upper Assam. AB - Current status of bancroftian filariasis among two distinct communities of Upper Assam was evaluated. High prevalence (10.3% mf rate) in tea workers descendents of tribals migrated from endemic states such as Orissa, Bengal, Uttar Pradesh and southern states of India, were recorded. Among non tea workers (indigenous population) significantly low mf rate was found in spite of the fact that both the communities are living in close vicinity and in similar ecological and mosquitogenic conditions. Culex quinquefasciatus was incriminated as vector in the areas of both the communities. A group of recent migrants of Bihar and Bengal living in the locality of non tea worker population were also found microfilaraemic. Significantly lower infection rate in non tea worker population appears to be associated with host susceptibility to the infection. Familial clustering in microfilaraemic subjects noticed in tea workers group also suggests involvement of racial susceptibility to infection. PMID- 10810597 TI - Bioactivity of ethanol extract of Karanja (Pongamia glabra vent) seed coat against mosquitoes. AB - Laboratory evaluation revealed that the treatment of larvae of Aedes aegypti and Culex quinquefasciatus with ethanol extract of karanja seed coat (ALKSC) significantly increased the larval mortality and developmental period proportianately with increase in the extract concentrations. Aedes proved more sensitive to the effect of extract in terms of mortality than Culex. At a high concentration (8 ppm), 100% Aedes larvae died in the first instar within two days whereas, 16 pmm was required for achieving the same level of mortality in Culex. Culex was also found more sensitive than Aedes regarding IGR effect. In emerging Culex pupae several abnormalities were observed while Aedes pupae did not exhibit any abnormalities. However larvae which pupated successfully resulted in adults with several structural abnormalities in both the species. PMID- 10810598 TI - Prevalence of Japanese encephalitis and West Nile viral infections in pig population in and around Chandigarh. AB - A study on sero prevalence of Japanese Encephalitis (JE) and West Nile (WN) viruses in pigs was carried out in and around Chandigarh. A total of 158 blood samples collected (June, 1995-March, 1996) from batches of pigs were screened for antibodies against JE and WN viral antigens by haemagglutination inhibition and complement fixation tests. In Chandigarh out of 13 pigs, 5 had JE and one WN HAI antibodies. Similarly, the sera of 14 and 1 pigs in Punjab and 29 and 3 pigs in Haryana were positive for JE and WN HAI antibodies respectively. In total JE specific HAI antibodies were found in 30.3% and CF antibodies in 12.5% of pigs whereas WN specific HAI antibodies were obtained in 3.2% and only one pig had CF antibodies. The results show that JE and WN viral infections are prevalent in pig population in and around Chandigarh. PMID- 10810599 TI - HIV infection in Varanasi. AB - A total of 6038 serum samples collected from individuals staying/residing in the eastern part of northern India, were screened for antibodies against HIV. The screening was done by ELISA and followed by confirmation of reactive samples by Western Blot. The seropositivity was detected in 154 (2.6%) persons. The prevalence amongst high risk groups was 68/1000 while it was 2.8/1000 in low risk population. The male to female ratio was 1.8:1 in seropositives. Increase by more than three folds in the prevalence was observed from 1994 (1.5%) to 1997 (5.0%). About 50% of the seropositives were in their third decade of life. Seropositives were predominantly (> 90%) from eastern Uttar Pradesh. Almost all the cases could be linked to Mumbai, one of the largest commercial and industrial centres of India. PMID- 10810600 TI - Role of macrophages in experimental malaria: VI--Effect of Freund's complete adjuvant in Plasmodium berghei infected mice. AB - Freund's complete adjuvant (FCA) treated group of mice when challenged with lethal Plasmodium berghei showed increased survival value; survival period (SP) and median survival day (MSD) compared to their respective control groups. K values were affected and mean parasitaemia during infection period was lower than that of control. In general survival rate after 35 days of infection was 10.5% in FCA recipients. The survival rate in a particular group of animals which received 0.2 ml FCA 3 days before challenge was 22.7%. FCA was found to contribute to increased survival of the host against P. berghei infection. The study indicates that adjuvants, like FCA induce protective immunity and future studies should include non-specific immunization against human malaria. PMID- 10810601 TI - Knowledge, attitude and practice towards malaria in rural communities of the epidemic-prone Thar Desert, northwestern India. AB - In order to assess the knowledge, attitude and practice of the rural Thar Desert population about malaria and its vectors, a total of 345 heads of households, including housewives, suffering with malaria were surveyed randomly, along with the same number of healthy control subjects, in four villages of Jodhpur and Jaisalmer districts of Rajasthan, north-western India. Results showed that in the traditionally hypoendemic Thar Desert, with low levels of immunity against malaria particularly Plasmodium falciparum, a large segment of rural people were equally less aware about the dangers of the disease. About three-fourths of the studied population did not quite comprehend the purpose of the five decade-old National Anti Malaria Programme, while nearly 50% respondents were reluctant to accept the programme lacking the component of community participation but directly targetting on the control of the disease. Consequently, about 50% malaria patients and 60% healthy subjects lacked in taking initiative to resort to any anti-mosquito or anti-malaria measure to protect themselves, although 27% malaria patients and 2.9% healthy subjects acknowledge correct causes of malaria. This study, being the first of its kind in a desert community currently facing emergence of P. falciparum-dominated malaria outbreaks, is a testimony to the hitherto prevalent deep gaps in knowledge about the disease, and its results warrant sincere efforts to link up the malaria control programme with the community participation for protection against this scourage in future. PMID- 10810602 TI - Cyclospora cayetanensis, a new emerging coccidian parasite. PMID- 10810603 TI - Prevalence of co-morbidity due to malaria: response to antimalarials. PMID- 10810604 TI - Prevalence of bancroftian filariasis in a foot-hill tea garden of upper Assam. PMID- 10810605 TI - Household characteristics and peoples' perception about filariasis in Khurda district Orissa. PMID- 10810606 TI - [Maintenance and termination of drug therapy in psychiatry]. PMID- 10810608 TI - [The 100th annual meeting of the Japan Surgical Society. Tokyo, Japan. April 12 14, 2000. Programs] [In Process Citation] PMID- 10810607 TI - [29th annual meeting of the Japanese Society of Neuropsychopharmacology. Hiroshima, Japan. September 14-15, 2000. Abstracts]. PMID- 10810609 TI - [The 100th annual meeting of the Japan Surgical Society. Tokyo, Japan. April 12 14, 2000. Abstracts]. PMID- 10810610 TI - Management problems in oncology. AB - Man has evolved sophisticated defence mechanisms over millions of years to combat insertion of foreign DNA into his cells. However, gene therapy carries huge potential for the treatment of cancer. The challenge is therefore to translate our scientific knowledge into a clinical reality. PMID- 10810611 TI - Adenoviral vectors. PMID- 10810612 TI - Retrovirus vectors. PMID- 10810613 TI - Targetable gene delivery vectors. AB - Adenoviral vectors, which have targeting ligands for tumor cells on the capsid, no natural tropism, and carry a therapeutic payload should be constructed soon and tested in pre-clinical models. Nevertheless, there are still important considerations for the design and therapeutic use of targetable vectors. Perhaps the single greatest challenge in the future, as it was in the past, will be finding ligands that have a higher apparent affinity for tumor and/or tumor endothelial cells then normal cells. However, the advent of many rapidly advancing technologies and information including the sequencing of the human genome, in vivo and in vitro phage display, rapid analysis of gene and protein expression in any context, and new cellular targets such as angiogenic endothelial cells, may provide many opportunities for the discovery of novel and useful ligands. In addition, the interests in targeting vectors are rapidly growing with new journals and meetings solely devoted to this subject increasing annually. Within the next 5 years, we should have meaningful clinical data on targetable vectors to reassess our progress. PMID- 10810614 TI - Human alpha-fetoprotein transcriptional regulatory sequences. Application to gene therapy. AB - The AFP regulatory sequences are among the best known tumor-specific transcriptional regulators. A number of groups have demonstrated that a variety of genes can be expressed in an HCC-specific manner under the control of the AFP regulatory sequences in vitro and in vivo. It would appear that, with the development of a suitable delivery system, HCC-directed gene therapy using the AFP regulatory sequences holds a promising future. PMID- 10810615 TI - Tumor-targeted Salmonella. Highly selective delivery vectors. AB - Genetically engineered Salmonella offer an intriguing new approach to selectively target solid tumors, including melanoma, lung, colon, breast, kidney and liver. These bacteria target tumors after systemic administration and selectively replicate within them. Specificity for tumors is often more than 1,000 times greater than for any other tissue. Auxotrophic mutations make these bacteria highly safe and form the basis for maintaining tumor specificity. An altered lipid greatly reduces the potential for septic shock yet also retains the antitumor properties of these bacteria. These bacteria have innate antitumor activity towards both primary and metastatic tumors and the ability to deliver proteins capable of activating chemotherapeutic agents directly within tumors. The delay in tumor growth results in mice that survive up to twice as long. These bacteria are susceptible to a wide range of antibiotics, allowing external control of the vector after administration. The combination of these features within a single vector seems specially surprising considering their unlikely source. PMID- 10810616 TI - Mutant adenoviruses selectively replication-competent in tumor cells. PMID- 10810617 TI - Polyoma and papilloma virus vectors for cancer gene therapy. PMID- 10810618 TI - Cochleates. Lipid-based vehicles for gene delivery-concept, achievements and future development. PMID- 10810619 TI - The use of skeletal muscle to express genes for the treatment of cancer. PMID- 10810620 TI - Adhesion molecules in cancer biology. PMID- 10810621 TI - Cell cycle control. PMID- 10810622 TI - KILLER/DR5, a novel DNA-damage inducible death receptor gene, links the p53-tumor suppressor to caspase activation and apoptotic death. AB - TRAIL and its emerging receptors are the newest members of the TNF receptor super family. The activation of TRAIL receptors by ligand binding leads to apoptosis through caspase activation through an as yet unclear signaling pathway that does not require the FADD adaptor. The TRAIL receptor KILLER/DR5, is induced by DNA damage and appears to be regulated by the tumor suppressor gene p53. Both the Fas receptor and KILLER/DR5 provide potential links between DNA damage-mediated activation of the p53 tumor suppressor and caspase activation. While further evaluation of the role of TRAIL receptors in human cancer is ongoing, initial studies suggest that both KILLER/DR5 and DR4 may be targets for inactivation and that these pro-apooptotic receptors may be tumor suppressor genes. Understanding the regulation of TRAIL and its receptors may thus be beneficial for the development of novel approaches for cancer treatment. TRAIL appears to be a cancer-specific cytotoxic agent and thus offers promise as a novel therapy for cancer either through replacement of the cytokine or potentially via gene replacement. Preliminary studies suggest the potential to combine TRAIL with classical cytotoxic chemotherapeutic drugs to achieve synergistic cell killing. PMID- 10810623 TI - Apoptin. AB - Tumor cells are often characterized by the traithey are more resistant to apoptosis induced by e.g. cytotoxic agents than normal cells. Resistance to apoptosis induction can be a direct consequence of mutations in certain tumor suppressor genes (p53) or of certain proto-oncogenes (Bcl-2). Therefore, new cancer therapies are under development to bypass the resistance to chemo- and radio-therapy of tumors. Apoptin acts independently of p53, is stimulated by Bcl 2 and is insensitive to BCR-ABL, which means that Apoptin can induce apoptosis in cases where present (chemo)-therapeutic agents, unfortunately, will fail. The fact that Apoptin induces apoptosis in human tumorigenic cells but not in normal diploid cells, implies that side-effects of Apoptin treatment are expected to be minor. In-vivo results with a first prototype of anti-tumor therapy based on expression of Apoptin indicate that Apoptin has low acute toxicity and is effective as an anti-tumor agent. PMID- 10810624 TI - Adenovirus-mediated herpes simplex thymidine kinase gene therapy for brain tumors. PMID- 10810625 TI - Development of clinical trial of E1A gene therapy targeting HER-2/neu overexpressing breast and ovarian cancer. PMID- 10810626 TI - Pre-clinical studies with tumor suppressor genes. PMID- 10810627 TI - Gene therapy for liver tumours. PMID- 10810628 TI - Adenovirus-mediated transfer of a p53 gene in ovarian cancer. AB - Given the lack of effective conventional therapy, those patients with recurrent or refractory ovarian cancer should be considered for currently approved investigational gene therapy protocols. Several studies have shown a potential modality of p53 gene transfer in cancer gene therapy. We also developed a new recombinant adenovirus carrying a wild-type p53 gene (AxCAp53). Although the efficacy of AxCAp53 to suppression of cell growth was not sufficient, AxCAp53 increased sensitivity to CDDP in ovarian cancer cells with deletion of the p53 gene. The combination of CDDP and AxCAp53 may be a potential strategy for the therapy of CDDP resistant ovarian cancer. PMID- 10810629 TI - Eliciting hyperacute rejection as a tumor killing strategy. Herpes amplicon vector transfer of the alpha(1,3)galactosyltransferase gene. PMID- 10810630 TI - Innate immune therapy for cancer. Screen for molecules capable of activating the innate immune system. AB - Tumor cells usually express antigens which are distinguishable from normal "self" antigens and are thereby recognized by the host immune system. However, the host immune system barely responds to tumors in patients. Supplementation with adjuvant (such as BCG-CWS) in patients with cancer contributes to regression of intrinsically growing cancer. The adjuvant targets antigen-presenting cells, i.e. innate immunity, but not lymphocytes, and promotes up-regulation of MHC, co stimulators and initial cytokines in antigen-presenting cells. We hypothesized that the role of the adjuvant is to provide conditions suitable for antigen presentation where antigens are available and the lack of adjuvant-induced priming of antigen-presenting cells results in unresponsiveness to tumor antigens. Here, we report innate immune therapy applicable to cancer patients by supplementation with adjuvants for induction of potent immune responses against tumors. PMID- 10810631 TI - Mda-7, a novel melanoma differentiation associated gene with promise for cancer gene therapy. PMID- 10810632 TI - Antisense IGF and antisense IGF-IR therapy of malignancy. PMID- 10810633 TI - Sensitization of tumors to chemotherapy through gene therapy. PMID- 10810634 TI - Design and testing of ribozymes for cancer gene therapy. AB - This chapter describes procedural aspects for development of ribozymes in general, and specifically, that cleave mRNA to an essential cellular gene, the AC40 subunit of RNA pol I. Ribozyme design includes functional selection of binding sites followed by computer modeling. These ribozymes are being used in vectors that target expression to the prostate via tissue specific promoters (Voeks, Norris, and Clawson, 1998) and have demonstrated efficacy. PMID- 10810635 TI - Ribozymes. Their design and use in cancer. PMID- 10810637 TI - Dendritic cell-based immunization for cancer therapy. PMID- 10810636 TI - The approach of triple helix formation in control of gene expression and the treatment of tumors expressing IGF-I. PMID- 10810638 TI - Hybrid cell vaccination for cancer immunotherapy. PMID- 10810639 TI - Modulation of the immune response through 4-1BB. PMID- 10810640 TI - Heat shock proteins in cancer therapy. PMID- 10810641 TI - Bi-specific antibodies in cancer therapy. PMID- 10810642 TI - B7.1 and cytokines. Synergy in cancer gene therapy. PMID- 10810643 TI - Intralesional vaccinia/GM-CSF recombinant virus in the treatment of metastatic melanoma. PMID- 10810644 TI - Approaches to gene-directed enzyme prodrug therapy (GDEPT). PMID- 10810645 TI - Suicide gene therapy. PMID- 10810646 TI - Adenoviruses as gene delivery vectors. PMID- 10810647 TI - Evaluation of an inhibitor of DNA methylation, 5-aza-2'-deoxycytidine, for the treatment of lung cancer and the future role of gene therapy. PMID- 10810648 TI - Vascular endothelial growth factor as a target for cancer gene therapy. PMID- 10810649 TI - Adeno-associated virus and other potential vectors for angiostatin and endostatin gene therapy. PMID- 10810650 TI - Potential applications of tissue inhibitor of metalloproteinase (TIMP) overexpression for cancer gene therapy. PMID- 10810651 TI - Vascular assessment. AB - The assessment of the patient with peripheral vascular disease encompasses a thorough history and physical examination with the adjunctive use of the noninvasive vascular laboratory to confirm, localize, and grade lesions. The need for additional vascular testing depends upon the clinical scenario and urgency with which intervention must be undertaken with contrast angiography reserved for planning surgical intervention. While multiple noninvasive and invasive methods are available to assess the peripheral vasculature, it should be obvious that not every patient requires an exhaustive battery of tests in order to evaluate their vascular status. In general, only those tests that are likely to provide information which will alter the course of action should be performed. Differing clinical syndromes mandate the extent of peripheral vascular testing. Table 5 lists distinct clinical syndromes and presentations and the peripheral vascular testing that is likely to yield timely and cost efficient results. PMID- 10810652 TI - Local flaps. AB - Local flap reconstruction of pedal defects can be accomplished with a variety of techniques. Multiple geometric constructs have been described for closure of foot defects. Each has its own unique principles which can be adapted to certain locations and premorbid conditions. Local flaps considered for closure of foot defects should lie within the higher level of any algorithm, due to their versatility, reproducibility, and long term functional outcomes. In addition, hospitalization, length of anesthesia, and perioperative care can be much less with the use of local flaps. PMID- 10810653 TI - Malignant melanoma of the foot and ankle. AB - Malignant melanoma is a serious and devastating skin disease that podiatrists may be called upon to treat. It is pertinent that delays in diagnosis and treatment of malignant melanoma be avoided. Some of the topics discussed in this article are causes, clinical features, classification, and treatment of malignant melanoma, focusing on the foot and ankle. PMID- 10810654 TI - Nerve injury associated with plantarflexion-inversion ankle sprains. AB - This article presents some of the less frequently described nerve pathologies associated with severe plantarflexion-inversion ankle sprains. It outlines the likely mechanisms of nerve injury, the typical presentations, and treatment possibilities. An anatomic review of the lower extremity, with emphasis on the neurologic structures, is also included to assist in understanding the mechanism and location of the nerve injury. PMID- 10810655 TI - Cimetidine and a delayed hypersensitivity reaction. AB - Oral cimetidine for the treatment of verruca continues to be a topic of discussion and controversy. Although its usage has gained popularity because it offers an alternative to conventional topical and surgical verruca therapy, its reported efficacy in managing warts lacks consistency in outcomes. Using high doses of oral cimetidine, therefore, raises concern about possible untoward effects. Cimetidine relies on effecting a change in the immune system to eradicate the verruca, but such a change in the immune system may cause patients to develop responses detrimental to their well being. The authors present an unusual case of severe delayed hypersensitivity with the use of oral cimetidine. PMID- 10810656 TI - [Dermatologic aspects of male hypogonadism]. AB - Since the skin and its appendages are androgen-dependent, male hypogonadism can be associated with dermatologically relevant lesions. The skin surface of hypogonadal patients is thin and shows increased wrinkles. In cases of prepubertal hypogonadism, severe acne does not occur. In contrast, patients may suffer from sebostasis due to missing stimulation of the sebaceous glands. The genital skin does not show pigmentation, which normally develops during puberty; the integument is pale. Hair growth on breast, pubia, axilla, face and integument is reduced; patients do not present with typical androgenetic alopecia. In addition to these features, dermatological diseases caused by treatment of the hypogonadism or drug-induced skin reactions have to be considered. While drug reactions to oral or intramuscular androgens or gonadotropins are rare, irritative or allergic skin lesions are observed frequently, especially in cases of non-scrotal transdermal therapeutic systems with testosterone. A variety of syndromes include dermatological symptoms associated with male hypogonadism; however, the prevalence of these syndromes is low. PMID- 10810657 TI - [Regression phase as therapeutic goal of cryosurgical treatment of growing capillary infantile hemangiomas. Treatment decision, treatment strategy and results of an open clinical study]. AB - BACKGROUND AND OBJECTIVE: Cryosurgery is a widely accepted modality for the treatment of capillary hemangiomas of the newborn. However, no studies clearly supporting its effectivity are available. PATIENTS AND METHODS: Out of 116 consecutive outpatients with 147 hemangiomas, 105 growing tumors in 91 patients were treated by cryosurgery. 13 further growing tumors were not treated because of lack of parental consent. Hemangiomas without growth within the last 2 weeks and with clinical signs of regression were not treated. After a follow-up of 1 year, the outcome of the treatment was classified as either regression (> 10% size reduction), no change (size alteration < 10%) or progression (> 10% size growth). The effectiveness of the treatment was assessed as clinical improvement of the hemangioma on an arbitrary scale per treatment session. RESULTS: Cryosurgery of growing hemangiomas efficiently initiated the regression phase (p < 0.05), in the most cases (68%) after a single treatment. Cryotherapy was most effective when employed in older children (p < 0.05) and for small hemangiomas (p < 0.05). Probably, this reflects associated spontaneous regression. CONCLUSIONS: Thus cryosurgery is a verifiably effective treatment modality for infantile capillary hemangiomas, shortening significantly their growth phase. PMID- 10810658 TI - [Contact allergies in patients with eczema of the external ear canal. Results of the Information Network of Dermatological Clinics and the German Contact Allergy Group]. AB - BACKGROUND AND OBJECTIVE: The Information Network of Departments of Dermatology (IVDK) centrally collects patch tests results in cooperation with the German Contact Dermatitis Research Group (DKG). Data of patients, who underwent patch testing to evaluate otitis externa were analyzed. PATIENTS/METHODS: Data of 145 patients that had been recorded between 1992 and 1997 were evaluated. The evaluation included data yielded by standardized patch testing as well as by patch testing of products that the patients had used. RESULTS: Allergic contact dermatitis was diagnosed in one third of patients suffering from otitis externa. Topical drugs were shown to be the most important allergens. In comparison to the entire IVDK collective, the population-adjusted frequency of sensitization (PAFS) of neomycin sulfate was higher in the study population whereas that of nickel sulfate was lower. That shows that in a very specific topographic area, different allergens may be of clinical significance. CONCLUSIONS: Whereas nickel sulfate represents an important allergen in the contact dermatitis of the external ear, topical therapeutical agents, especially neomycin sulfate and, as evaluation of the patch tests of the patient's own products revealed, probably polymyxin B are the dominating allergens in otitis externa. PMID- 10810659 TI - [Transitory acantholytic dermatosis (Grover disease). An analysis of the clinical spectrum based on 21 histologically assessed cases]. AB - BACKGROUND AND OBJECTIVE: In 1970 Ralph W. Grover described a clinicopathologic entity characterized by pruritic keratotic papules and papulovesicles predominantly on the trunk, disappearing spontaneously after a few weeks or months and demonstrating the histological features of epidermal acantholysis. The etiology remains unknown; sweating, heat and sunlight are suspected trigger factors. In our survey the clinical spectrum of the disease will be examined. PATIENTS AND METHODS: We have analyzed 21 cases of Grover's disease histologically diagnosed in Berne in 1997 and 1998. RESULTS: In most cases we observed isolated papules disseminated on the trunk. The predominant histological type was the Darier-type; presumably there is no correlation between histological type and clinical features. At the time of diagnosis the skin disorder had been present on average for 83 months. There was a strong association with sweating and heat. Topical steroids were successful in 50% of patients. CONCLUSIONS: Our 21 cases reflect in general the literature. Nevertheless, the long persistence of the skin lesions was striking perhaps making the term transient somewhat inaccurate. PMID- 10810660 TI - [Sjogren-Larsson syndrome]. AB - This rare, ubiquitous neurocutaneous disorder is inherited in an autosomal recessive fashion. Its primary clinical manifestations are congenital ichthyosis, spastic diplegia or tetraplegia, and mental retardation. The causative biochemical defect has been identified as a deficiency of the enzyme fatty aldehyde dehydrogenase, a component of fatty alcohol:NAD+ oxidoreductase. We present a case report of an affected 3.5 year old white girl to give an overview of the pre- and postnatal diagnostic procedures as well as of therapeutic options. PMID- 10810661 TI - [Specific skin infiltration of extracutaneous B-cell lymphoma in healing herpes simplex skin]. AB - Both herpes simplex and herpes zoster are not uncommon in patients affected by malignant lymphoma or leukemia. Of particular interest, the herpetic lesions may be followed by specific cutaneous infiltrates. We report on a patient with a centrocytic-centroblastic non-Hodgkin lymphoma who showed primary skin infiltrates within the area affected by herpes simplex. After effective therapy with aciclovir, papules and plaques developed at the site of the herpetic scars. Histologic investigation confirmed dense B-lymphocytic infiltrates with immunoglobulin light chain restriction and a granulomatous reaction. Although the chemotherapeutical regiment was modified, the patient died one month later of a lethal blast crisis. The occurrence of so called isotopic skin reactions and their prognostic significance is discussed. PMID- 10810662 TI - [Squamous epithelial carcinoma in erythema ab igne]. AB - Erythema ab igne (EAI) is a red-brown net-like hyperpigmentation due to repetitive exposure to submaximal heat over a long period of time. This rash is typically seen on the anterior lower legs of people who sit in front of open fireplaces or coal stoves and on the trunk of patients with chronic disorders who seek pain relief by long-term use of hot-water bottles or heating pads. EAI can not only be a diagnostic sign for underlying internal diseases including malignant tumors but can also give rise to cutaneous malignancies. We report on a 75-year-old female patient who developed several thermal keratoses and squamous cell carcinomas at the site of erythema ab igne caused by chronic heat exposure over several years. PMID- 10810663 TI - [Erythema palmare hereditarium ("red palms")]. AB - We report a woman with erythema palmare hereditarium, an anomaly not documented so far in the German literature. It is characterized by a bright erythema of the palms, usually persistent since birth. It is transmitted in an autosomal recessive mode and has a benign course. In our patient it was possible to trace the palmar erythema over three generations. PMID- 10810664 TI - [Pili trianguli et canaliculi. A case report of uncombable hair in relation to atopic eczema and tooth anomalies]. AB - We report on a child with pili trianguli et canaliculi. This hair shaft abnormality belongs to a heterogeneous group of diseases which are included under the synonym uncombable hair. The diagnosis was confirmed by scanning electron microscopy, revealing hair shafts with a characteristic longitudinal groove. In addition the girl suffered from atopic eczema and tooth anomalies. Our findings suggest that this disorder could represent a tricho-odontal subtype of ectodermal dysplasia. PMID- 10810665 TI - [Annular erythema in Sjogren syndrome. A variant of cutaneous lupus erythematosus?]. AB - Annular erythema has been recognised as a cutaneous manifestation of Sjogren's syndrome in the Asian literature and has been assumed to represent a distinct clinical entity. Since there are common pathophysiologic mechanisms, mainly the presence of anti-Ro/SSA or anti-La/SSB antibodies, it is difficult to separate the annular erythema from subacute cutaneous lupus erythematosus. Histological examination may reveal dermal mucin deposition resembling lupus erythematosus tumidus. We present a Chinese patient with widespread annular erythema, keratoconjunctivitis sicca, and anti-Ro/SSA antibodies. Clinical, histopathological, and immunogenetic findings are discussed reviewing the current literature, and the differences between annular erythema associated with Sjogren's syndrome and cutaneous lupus erythematosus are emphasized. PMID- 10810666 TI - [Alternative prescriptions]. PMID- 10810667 TI - [Success of photodynamic therapy in circumscribed scleroderma. Interview with PD Dr. M. Szeimis, Regensburg University Dermatologic Clinic]. PMID- 10810668 TI - [Renal failure after eczema treatment with Chinese herbal medicine]. PMID- 10810669 TI - [In Process Citation] PMID- 10810670 TI - [Effectiveness of totally implantable auditory prostheses in inner ear deafness]. PMID- 10810671 TI - [Electrodiagnosis and decompression in idiopathic facial nerve paralysis. A view of the maze of conflicting opinions]. PMID- 10810672 TI - [Treatment of the hypertrophied inferior turbinate. 2]. AB - The surgical management of enlarged inferior turbinates has been actively debated for more than a century. The second part of this review describes the following techniques: laterofracture, submucous turbinectomy, concho-antropexy, partial turbinectomy, anterior turbinectomy, inferior turbinoplasty, total turbinectomy, and vidian neurectomy. Indications, advantages, disadvantages, complications, and controversies of each treatment modality are reviewed and discussed. Partial or submucosal resection should be preferred, because there are moderate side effects and the amount of turbinate excised can be altered according the degree of symptomatology. Total turbinectomy or vidian neurectomy are indicated if all other treatment attempts do not succeed. PMID- 10810673 TI - [Temporal inconsistency of neuromyography of the facial nerve in healthy probands]. AB - Facial neuromyography is an objective electrophysiologic measurement used to assess prognosis for recovery from facial paralysis. In the present study variability over time of the amplitude ratio in healthy subjects was investigated. Twenty healthy subjects underwent four neuromyographic tests at intervals of one week. The study group consisted of 12 males and 8 females. Ages ranged from 23 to 36 years. The stimulation intensity required for supramaximal stimulation was 14.6 mA on average. The mean of the intra-individual side-to-side difference of the evoked compound action potential was 32.5%. Results of the four repeated tests ranged from no to 80% side-to-side differences in the same individual. The hypothesis of symmetrical evoked facial compound potentials was not supported by our data. As with previous investigators we found significant side-to-side differences in healthy subjects. Furthermore, we could also show for the first time that the amplitude ratio was not constant in every individual at repeated measurements. We conclude that there is a physiological variability that must be taken into account in the interpretation of facial nerve neuromyography. PMID- 10810674 TI - [What is the value of the Denecke surgical therapy concept in deglutition disorders?]. AB - BACKGROUND: Denecke developed 1957 a surgical strategy consisting of three or four operative steps for rehabilitation of patients with unilateral paralysis of IXth and Xth cranial nerve. These four steps are the following: myotomy of the cricopharyngeal muscle, resection of the paralyzed pharyngeal wall, medialization of the paralyzed vocal cord and fixation of the soft palate to the posterior pharyngeal wall which can be facultative. PATIENTS AND METHODS: For the first time we have reviewed patients operated on with the Denecke operation in a retrospective study. The surgical results should be analyzed to deduce some parameters for surgical indication. RESULTS: 16 patients were reviewed in regard to nourishment status, situation of tracheostomy, former pneumonias, classification of Miller and Eliachar for aspiration and subjective evaluation. The follow-up was 21 +/- 16 months postoperatively. In 8 cases surgical intervention was successful. In 2 additional cases the results were partially satisfying. CONCLUSIONS: The surgical procedure of Denecke was successful for improving substantially nourishment and aspiration status of considerably reduced patients in about 50% of the reviewed cases. Preoperative cinematography of swallowing, manometry and an interdisciplinary approach are required to clearly define the indications for surgical intervention. PMID- 10810675 TI - [Clinical comparison of a digital with an analog hearing aid]. AB - The benefit of innovative hearing aid technology can be evaluated in clinical trials. The present study describes the comparison of a new digital hearing aid with an analogue device serving as a reference. The tests were carried out with 15 experienced hearing aid wearers. To prevent the influence of different fitting algorithms such as prescriptive or loudness-based the digital devices were not renewed fitted but their level- and frequency-depending gain was adjusted to that of the reference. Different tests concerning loudness perception (category loudness scaling) speech discrimination in noise (the Gottingen sentence test) and self-assessment of the benefit by questionnaires were performed. All tests yielded slightly better results for the digital hearing aid. The speech audiometric evaluation showed somewhat better discrimination for the test-device. The questionnaires yielded a marked preference for the digital device. Because the study was not blinded influences due to the knowledge of the subjects of testing a new technology generally can occur. On the other hand, the question arises whether present audiometric tests sufficiently consider signal processing of modern hearing systems. Furthermore, one has to take into account that only a few of the features of digital technology such as noise-reduction and feedback cancellation were considered in this study to allow for a sensible comparison. Because of the large number of possibilities offered by digital technology additional benefit by the hearing aid can be expected. PMID- 10810676 TI - [Vestibulospinal reactions in cervicogenic disequilibrium. Cervicogenic imbalance]. AB - A functional cervical spine disorder is often the cause for persistent vertigo, which can last months or several years. The existence of cervical vertigo is not generally recognized, mainly because an objectivation of the cervical nystagmus is not easily understood by many examiners. In this study we examine additional parameters, which underline the diagnosis of cervical imbalance. The anamnestic statement of staggering refers to a disturbance of the vestibulospinal reactions. In 67 patients in which cervical imbalance was suspected the vestibulospinal reactions were monitored directly before and after manual therapy of the cervical spine. The cranio-corpo-graphie (CCG) and the posturography were used to monitor the results. A highly significant improvement of pathological vestibulospinal reactions was seen after chiropractic manipulation of the spine. These results show that a functional disorder of the cervical vertebrae influences the vestibulospinal reactions. The pathological deficit of the vestibulospinal reactions is not solely a phenomenon of peripheric labyrinth malfunction, failure in the brainstem or in the area of the cerebellum ("brain stem staggering"), but can also be viewed nearly regularly by cervical disturbance of the equilibrium. The results of the treatment can be observed within a few hours. PMID- 10810677 TI - [Use of an intraoral snoring therapy device of thermolabile plastic]. AB - Oral appliances are used to treat snoring and sleep apnea. Yet, their success cannot be predicted without a therapeutical trial. This uncertainty and the high prices of the appliances are the reasons for their limited use. We tested a cheap, custom fit mandibular advancement device (SnorBan) for the treatment of sleep disordered breathing in order to assess its efficacy. 39 consecutive patients (51.1 +/- 9.2 years, BMI = 27.4 +/- 4.5 kg/m2) with different degrees of sleep disordered breathing (AHI = 16.6/h +/- 15.6/h) received the device after a thorough clinical examination. After getting used to the device a second polysomnography was performed. The AHI improved significantly from 16.6/h to 8.2/h (P < 0.01) in the whole group. The only patient who became worse could not get used to the device. Time with snoring dropped significantly from 16.3% to 6.6%, 59.1% of the sleep apnea patients were successfully treated as their RDI dropped below 10/h. The sleep efficiency remained unchanged. Slow wave sleep and REM-sleep increased significantly from 12% to 16% (P < 0.05). The overall compliance was 75%. The custom fit mandibular advancement device Snorban is a cheap and effective treatment for a number of patients with snoring and sleep apnea. The oral appliance is proven to be a useful and simple, non-surgical treatment option. Polysomnographic follow-up is mandatory as breathing may worsen with the device while asleep. PMID- 10810678 TI - [Acceptance of wearing a hearing aid by children]. AB - BACKGROUND: Having one or 2 hearing aids is not a guarantee that a child will wear the instruments constantly. The purpose of this study was to evaluate the hearing aid acceptance by children (assessed using the daily wearing time). PATIENTS AND METHODS: A total of 116 parents of a patient population of persistent hearing-impaired children with a need for a hearing aid (average age 51.1 +/- 30.1 months) rated the wearing acceptance of their children on a 5-point Likert-scale. This assessment was made after the child had received its individual hearing aid for its specific disorder (on average after 5 months). Hearing levels ranged from mild to profound. RESULTS: 58.6% of the children accepted their electroacoustic aids amplier "excellent" or "good", 18.1% did it "average", 23.3% "bad" to "miserable" or even "not at all". There was no significant difference between girls and boys. The wearing acceptance from children with unilateral hearing impairments was reduced, especially from those with chronic conductive disorders by malformations of the ears. Very young children (prelingual hearing impairments), mainly those with additional handicaps and/or not German speaking homes demonstrated a "bad" to missing wearing behaviour. Hearing acceptance did not correlate with the severity of the disorder. CONCLUSION: The wearing acceptance as one aspect of compliance should be rated before each regular hearing aid control. Finally it is an efficient and sensitive mean to detect faulty amplification and changed diagnosis. PMID- 10810680 TI - [Highly malignant T-cell non-Hodgkin lymphoma (nasal type) of the hard palate]. AB - The monomorphic clinical aspect of destructive mid-face lesions is characterised by inflammation, induration and granulomatous transformation. This feature can be caused by various infections, toxical noxa, Wegener's Granulomatosis and different neoplasms. The case of a 19 year old patient with EBV associated nasal type T-cell lymphoma located at the hard palate is presented. The diagnostic approach and difficulties in diagnosing this entity assessing by using multiple biopsies, serological and molecularbiological detection of EBV association and immunohistochemistry for atypic T-cells are elucidated. In the presented case the treatment with chemotherapy and irradiation following a well-defined therapy concept leaded to a three year recurrence-free survival so far. The comparison of the key-histological findings and the major differential diagnoses is mandatory to establish the final diagnosis of lymphoma. This is the basement for treating this disease with combined chemotherapy and irradiation for optimizing survival. PMID- 10810679 TI - [Identification of a nasopharyngeal carcinoma using Epstein-Barr virus diagnosis]. AB - We report the case of a 36-year-old women who was found to have a malignant tumor extending from the side of her right nasal cavity to the nasopharynx. Magnetic resonance imaging and computed tomography were not able to define the primary site of the tumor. Histologic evaluation demonstrated an undifferentiated squamous cell carcinoma. Because of the different treatment concepts for carcinomas of the nasal cavity and nasopharynx, we tried to identify the primary site by diagnosing Epstein-Barr virus (EBV) infection, which is associated with carcinoma of the nasopharynx. By using immunohistochemistry and polymerase chain reaction EBV could be identified in the cells of the carcinoma. This showed that the primary site of the tumor was located in the nasopharynx and resulted in the patient being treated with simultaneous radiochemotherapy. PMID- 10810681 TI - [Emergency management in ENT general practice. 1: Basic measures]. PMID- 10810682 TI - [Multiple foreign body aspiration]. PMID- 10810683 TI - [B-image sonography. 1. Physical principles]. PMID- 10810684 TI - Neer Award 1999. Overuse activity injures the supraspinatus tendon in an animal model: a histologic and biomechanical study. AB - Overuse activity has been implicated as an etiologic factor in injury to the rotator cuff and to the supraspinatus tendon in particular. Due in part to the lack of an appropriate animal model, expex85ental studies have not addressed this issue. With the use of a rat model, we measured the effects of an overuse running regimen on 36 Sprague-Dawley rats after 4 (n = 12), 8 (n = 12), or 16 (n = 12) weeks of exercise and compared them with a control group of rats (n = 10) who were allowed normal cage activity. The histologic characteristics, the gross morphologic characteristics, and the mechanical properties of the tendon tissue were evaluated. The supraspinatus tendons in the exercised animals demonstrated significant changes as a result of overuse at all time points compared with the normal group. There was an increase in cellularity and a loss of the normal collagen fiber organization consistent with what has been seen in human tendinopathy. The tendons from the exercise groups were larger than normal in cross-sectional analysis at 4 weeks (129% of control, P < .01) and continued to increase in size with time to 16 weeks (164% of control, P = .01). The mechanical properties of the tendons deteriorated in response to overuse exercise with a decreased modulus of elasticity ranging from 52% to 61% of control (P = .07 at 4 weeks, P < .05 at 8 and 16 weeks) and a decreased maximum stress of failure ranging from 51% to 63% of control (P < .007). These findings support overuse activity as an etiologic factor in the development of supraspinatus tendinopathy and begin to describe the changes in the tendons as a result of such activity. This model can now be used to study the effect of various treatment modalities on these injuries. PMID- 10810686 TI - Open surgical treatment for dialysis-related arthropathy in the shoulder. AB - The clinical results of open surgical treatment for hemodialysis-related arthropathy in the shoulder were evaluated, and the pathology was investigated. Fifteen shoulders of 11 patients (6 female and 5 male) were treated during the past 6 years. The average age was 56.7 years. The duration of hemodialysis averaged 17.5 years. Simple curettage was performed in 7 shoulders, and curettage and bone graft were performed in 5 shoulders. Repair of the rotator cuff was performed in 3 shoulders. Pathologic foci always existed in the intertubercular sulcus and the greater tuberosity of the humerus. Marked proliferations of the bursa and multiple cystic bone lesions were found, and amyloid depositions were observed on histologic evaluation in all cases. Open surgical treatment was quite effective, and relief from the clinical symptoms was maintained for a satisfactory period of time. Enthesopathy induced by the amyloid deposition is considered the main pathology. PMID- 10810685 TI - Patient-controlled lidocaine analgesia for acromioplasty surgery. AB - Twenty-four consecutive patients undergoing shoulder acromioplasty were given postoperative analgesia with a new method in which a patient-controlled continuous infusion of lidocaine infiltrated the subacromial space. Seventeen of the acromioplasties were done with arthroscopy, whereas 7 were performed with an open procedure. A 2% solution of lidocaine without epinephrine was used for both a continuous dose of 2 cc/h and patient-controlled interval doses of 1 cc administered at 15-minute intervals. The catheter was left in place for 72 hours. We prospectively studied complications, the patient's subjective pain level, the amount of supplementary pain medication used, and serum levels of lidocaine. In addition, we evaluated a control group of 24 patients undergoing acromioplasty by the same surgeon without the use of this method of pain control. No wound complications occurred. No adverse reactions to lidocaine or overdose of lidocaine occurred. Blood levels of lidocaine averaged 0.3 microgram/mL in the 12 patients studied. Subjective pain levels and the amount of supplementary pain medication used were both lower in the group receiving patient-controlled lidocaine analgesia at statistically significant levels (P = .168 measuring subjective pain level, and P = .0212 measuring supplementary pain medication use). Patient-controlled lidocaine analgesia in the subacromial space appears to be a safe method for achieving high levels of pain control in patients undergoing an acromioplasty. PMID- 10810687 TI - Absorbable implants for open shoulder stabilization: a clinical and serial radiographic evaluation. AB - PATIENTS AND METHODS: Eighteen consecutive patients who had recurrent, unidirectional, post-traumatic shoulder instability were included. All the patients underwent surgery with a standardized open Bankart technique involving absorbable suture anchors. CLINICAL RESULTS: No redislocations occurred during the study period of 31 months (range 25 to 38 months). The Rowe and Constant scores were 86 points (range 61 to 98 points) and 89 points (range 73 to 99 points), respectively. The strength measurements revealed 8.6 kg (range 3.8 to 15 kg) in 90 degrees abduction compared with 9.3 kg (range 2.2 to 16.5 kg) in the control shoulders (not significant). The external rotation in abduction was 65 degrees (range 20 degrees to 90 degrees) compared with 91 degrees (range 80 degrees to 105 degrees) in the control group (P < .001). RADIOGRAPHIC RESULTS: Signs of minor or moderate degeneration were found in 10 of 18 patients on the preoperative radiographs, in 15 of 18 at 7 months, and in 16 of 18 at 33 months (P < .05 before surgery vs 33 months). From the preoperative examination to the 7 month control, 7 of 18 patients had an increase in degenerative changes, and from the 7-month to the 33-month control, an increase was found in 8 of 18 (P = .008, before surgery vs 7 months; P = .005, 7 months vs 33 months). At the 7-month control, 8 of 18 patients had invisible or hardly visible drill holes, and 10 of 18 had visible or cystic drill holes at the site of implantation for the absorbable suture anchors. At the 33-month control, 10 of 18 patients had invisible or hardly visible drill holes, and 8 of 18 had visible or cystic drill holes (not significant, 7 months vs 33 months). CONCLUSION: The method resulted in stable shoulders in 17 of 18 patients. Degenerative changes were present on the radiographs in most of the patients and appeared to increase over time. Visible drill holes or drill holes with cystic changes were seen on the radiographs in a significant number of patients at the 7-month and the 33-month control and did not appear to heal during the follow-up period. PMID- 10810688 TI - Parsonage-Turner syndrome in the Native American Indian. AB - Neurologic causes are important to consider when patients with new-onset shoulder pain are evaluated. Parsonage-Turner syndrome refers to a specific clinical form of brachial plexus neuropathy, with most evidence indicating an infectious or immune cause. In the United States reports have been limited to reviews of sporadic cases despite epidemiologic clustering in other geographic locations. This report presents an outbreak in the Native American Indian population from the southwest United States, representing the first epidemiologic cluster in this country. This occurrence exhibits a distinctive expression of the essential clinical features, a clear association with antecedent febrile illness, and an occurrence in an ethnic population. As diagnosis continues to be based on careful clinical observation, surveillance by knowledge of the disease will shed further insight into pathogenesis. Accordingly, Parsonage-Turner syndrome should be included during the diagnostic evaluation of patients with new-onset shoulder pain. PMID- 10810689 TI - Shoulder prosthesis subluxation: theory and experiment. AB - The articular shapes of the humeral and glenoid components in total shoulder arthroplasty affect the loading, translation, and contact stresses in the joint, thereby affecting stability, glenoid loosening, and wear. Experiments were conducted to determine the subluxation load and corresponding translation for 6 types of glenoid components. The effects of shape, size, testing direction, compressive load, testing speed, testing medium, bone substitute properties, and repeated subluxations were investigated and compared with theoretical, rigid-body predictions. The subluxation load, varying from 45% to 98% of the axial load for the prostheses tested, is affected by glenoid constraint (i.e., the maximum slope at the glenoid articular rim), the compressive load, the coefficient of friction, and the deformability of the articular edge. Rigid-body theory overestimated the experimental load, which was not surprising given the visible deformations, but provides a framework to highlight the relevant design parameters. The subluxation translation, ranging from 1 to 13 mm for the prostheses tested, is determined by the glenoid length and by the conformity between the humeral and glenoid radii. Experimental translations were greater than rigid-body predictions for the most conforming prostheses and roughly equal for less conforming prostheses. The goals of this study were to characterize the subluxation load and translation of a variety of types of prostheses, to develop the rigid-body basis for these results, to compare the rigid-body and experimental results, and to locate experimentally the glenoid articular rim for further testing. PMID- 10810690 TI - Axillary view of the glenoid articular surface. AB - The glenoid articular surface is best studied radiographically on a tangential projection with both true anteroposterior and axillary views. Forty-one dry scapulas were studied under fluoroscopy to define the axillary projection that would provide a true scapular lateral view. The superior and inferior articular margins were marked with radiopaque solder wires. A true axillary view was obtained when the projection of the wires superimposed. The projection of the cortical bone of the posterolateral surface of the coracoid was noted to be continuous with the projection of the subchondral cortical bone of the glenoid articular surface when the latter was viewed tangentially. An illustrative case is shown in which a screw was mistakenly seen violating the glenohumeral joint; however, with the defined true axillary view, the actual position of the screw was demonstrated. X-ray films taken of another 8 cadaver shoulders were used to study the position of screws inserted about the glenoid articular surface. The soft tissue shadow superimposition on the inferior glenoid margin can lead to a misinterpretation of the superior margin as the whole glenoid articular surface. Because soft tissue can interfere with the appreciation of the glenohumeral joint line on an axillary view, a projection that will show a continuous line of the coracoid and glenoid articular surface should be obtained, and it will indicate a tangential view of the joint. PMID- 10810691 TI - Repair of distal biceps tendon rupture: a new technique using the Endobutton. AB - The authors describe a technique with a single anterior incision and fixation with an internal button, the Endobutton. The procedure is performed through a 5 cm transverse skin incision, and the tendon is sutured to the Endobutton with 2 number 5 Ethibond sutures. Surgical repair in the depths of the muscular forearm is not required, because the tendon is simply sutured external to the wound. The Endobutton delivers and locks the tendon into a hole in the radial tuberosity. The Endobutton technique was used in 12 patients who were allowed early active mobilization. All were satisfied, returned to activities, and regained grade 5 strength. Average flexion was from 5 degrees to 146 degrees with 81 degrees supination and 80 degrees pronation. No neurovascular complications or synostosis occurred. In cadaveric studies the average distance from the biceps tendon were ulnar artery 6 mm, median nerve 12 mm, and posterior interosseous nerve 18 mm. The average distance from the posterior interosseous nerve to a Steinman pin advanced through the proximal radius was 14 mm. This technique is a safe and effective method of repair of distal biceps tendon avulsion that allows active mobilization with minimal risk of complication. PMID- 10810692 TI - Kinematics and laxity of the Souter-Strathclyde total elbow prosthesis. AB - The motion pattern and laxity of 8 cadaveric elbows were recorded with a 3 dimensional electromagnetic tracking device before and after the Souter Strathclyde total elbow prosthesis was implanted. The Souter-Strathclyde prosthesis replicates the valgus-varus motion pattern of the intact elbow but causes a significant internal rotation of the ulnar shaft of 8.9 degrees +/- 4.1 degrees (P < .0005) at 110 degrees of elbow flexion. One of the reasons for this unphysiological motion pattern is positioning of the humeral component in a mean of 5.4 degrees +/- 6.4 degrees of external rotation compared with the intact elbow (P = .05). This positioning is related to the design of this device. The Souter-Strathclyde elbow prosthesis has a mean maximum valgus-varus laxity of 6.5 degrees +/- 1.5 degrees compared with 4.3 degrees +/- 2.3 degrees for the intact elbow (P = .004). This implant is more constrained than previously tested devices, which may explain its relatively higher loosening rate. PMID- 10810693 TI - Total elbow replacement with the GSB III prosthesis. AB - Fourteen consecutive elbows have been treated for rheumatoid arthritis (9 elbows) and for post-traumatic osteoarthrosis (5 elbows) by total elbow replacement with the GSB III implant. The elbows were reviewed retrospectively after a mean follow up of 6 years (2 to 9 years). Ten of 14 elbows had a functioning GSB III implant at follow-up; 7 of them were rated satisfactory and 3 unsatisfactory with the Mayo elbow performance score. In 5 of these 10 cases, the patients had significant pain relief with no or only mild pain at follow-up, whereas 5 had moderate to severe pain. With a functioning implant the range of motion averaged 140 degrees of flexion, 19 degrees of lack of extension, 65 degrees of pronation, and 84 degrees of supination. Six (43%) elbows had major complications requiring 1 to 8 additional operations. Aseptic loosening requiring revision occurred in 4 (29%) elbows. Two of them were treated by a resection arthroplasty, and 2 were revised with another hinged semiconstrained device. Three further elbows had radiolucent lines involving more than 50% of the cement-bone interface of either the humeral or the ulnar component. However, in 8 elbows the cementing technique was considered marginal or inadequate. Poor cementing (marginal or inadequate) was associated with loosening (P = .008). The GSB III total elbow prosthesis can restore function and reduce pain. The rate of aseptic loosening in this series was higher than previously reported. Based on this observation, we conclude that the GSB III implant seems to be sensitive to the insertion technique and does not tolerate suboptimal cementing. PMID- 10810694 TI - Intraosseous capture of the long bicipital tendon after fracture-dislocation of the shoulder: a spontaneous Nicola procedure. PMID- 10810695 TI - Winging of the scapula caused by disruption of "sternoclaviculoscapular linkage": report of 2 cases. PMID- 10810696 TI - Ilizarov reconstruction after a failed upper limb salvage procedure in oncology: a case report. PMID- 10810697 TI - Acute compartment syndrome of the supraspinatus: a case report. PMID- 10810698 TI - Juxta-articular myxoma of the shoulder presenting as a cyst of the acromioclavicular joint: a case report. PMID- 10810699 TI - Iatrogenic ulnar nerve injury after percutaneous cross-pinning of supracondylar fracture in a child. AB - Supracondylar fracture of the humerus is the most common fracture of the elbow in children and has been treated by a variety of methods. Recently, stabilization of reduced fractures with percutaneous pin fixation has become the accepted method of treatment. Ulnar nerve injury is a complication of percutaneous pinning of supracondylar fractures, although many authors have reported that it resolves spontaneously after removal of the pin. PMID- 10810701 TI - Synthesis of 6,8-diazabicyclo AB - [formula: see text] Starting with the proteinogenic amino acid (S)-glutamate, a general method for the synthesis of 3-(piperazin-2-yl)propionic acid esters 7 with various substituents at N-4 of the piperazine ring system is presented. An intramolecular ester condensation of 7 is the key step in the formation of the 6,8-diazabicyclo[3.2.2]nonane derivatives 8-10, which are of interest as conformationally restricted piperazines. PMID- 10810702 TI - A chiral 2-D donor-acceptor array of a bipyrazine N-oxide and tetracyanoethylene AB - [formula: see text] Hexamethylbipyrazine-N,N',N",N"'-tetraoxide (1) is synthesized in racemic form and cocrystallized with tetracyanoethylene to give a donor-acceptor (DA) networked lattice. The resulting DA2 cocrystal contains homochiral 2-D DA arrays (layers) cohered by 2.7 A NO...C=C DA bonds with periplanar O p-orbital/C=C orientations. Layer formation is stereoselective, while interlayer relations yield a racemic lattice. PMID- 10810700 TI - Partial-thickness rotator cuff tears: a modern view on Codman's classic. PMID- 10810703 TI - A synthetic hydroxy acid that shows tubular-shaped structure in solid-state and ionophoric activity in phospholipid bilayers AB - [formula: see text] In this contribution, we describe the ability of compound (+/ )-1b and six molecules of water to form in solid-state hexameric aggregates, which self-assemble to give hollow tubular structures. Single-crystal X-ray analysis shows that these tubes are open-ended, with irregular shape and internal van der Waals pore diameter between 6 and 9 A. In addition, transmembrane sodium transport activity was also assessed for (+/-)-1b using dynamic Na(+)-NMR technique. PMID- 10810704 TI - Linear free energy relationships implicate three modes of binding for fluoroaromatic inhibitors to a mutant of carbonic anhydrase II. AB - [figure: see text] Linear free energy relationships between binding affinity and hydrophobicity for a library of fluoroaromatic inhibitors of F131V carbonic anhydrase II (CA) implicate three modes of interaction. X-ray crystal structures suggest that F131 interacts with fluoroaromatic inhibitors, while P202, on the opposite side of the active site cleft, serves as the site of the hydrophobic contact in the case of the F131V mutant. 2-Fluorinated compounds bind more tightly, perhaps due to the field effect of the nearby fluorine on the acidity of the amide proton. PMID- 10810705 TI - A solution to the cyclic aldol problem AB - [formula: see text] A protocol for achieving stereoselective aldol reactions with cyclic ketones is presented. In terms of yield, the process is particularly effective when a quaternary center at the alpha-carbon of the beta-hydroxy ketone product is created. The stereochemical outcome, anti or syn, is achieved by the Lewis acid-mediated ring expansion of stereochemically homogeneous epoxides in a reaction related to the pinacol rearrangement. PMID- 10810706 TI - Preparation of benzocycloheptene derivatives from zirconacyclopentadienes AB - [formula: see text] Reaction of zirconacyclopentadienes with 2-iodobenzyl halide and 2-iodobenzoyl halide afforded benzocycloheptene derivatives and benzocycloheptenone derivatives in good to high yields. PMID- 10810707 TI - Formation of new 5,11b-bridged isoindolo AB - [formula: see text] The tandem Pummerer/pi-aromatic cyclization of alpha acyliminium ion, leading efficiently in "one pot" to thiopoxyareno-bridged isoindoloisoquinolinone incorporating the arylthio group, has been demonstrated for the first time. During this sequence the angular hydroxylated isoindoloisoquinolinone, resulting from the Pummerer-type cyclization, was also obtained. PMID- 10810708 TI - Unexpected products via singlet oxygen oxygenation of functionalized 5,6-dihydro 1,4-oxathiins AB - [formula: see text] Single oxygen oxygenation of 5,6-dihydro-1,4-oxathiins substituted at C-3 with an electron-withdrawing group leads stereoselectively to ketosulfoxides 5 and 6, instead of the expected dicarbonyl compounds 3. A mechanism involving an unprecedented intramolecular rearrangement of the corresponding dioxetanes 2 is proposed. PMID- 10810709 TI - Stereoselective synthesis of skipped polyols by substrate-directed asymmetric induction AB - [formula: see text] A series of C2- or sigma-symmetric oligo-1,3 dioxanylmethanes, 2-5, have been prepared using a bidirectional approach. In bidirectional syntheses of meso compounds, only substrate-based asymmetric induction could be applied. 1,3-Asymmetric induction in Mukaiyama-aldol additions, 1,5-asymmetric induction in enol-borinate aldol reactions, and 1,3 anti-selective reduction of aldols turned out to be reliable tools in the preparation of compounds 3-5. PMID- 10810710 TI - Suppressed beta-hydride elimination in palladium-catalyzed cascade cyclization coupling reactions: an efficient synthesis of 3-arylmethylpyrrolidines AB - [formula: see text] A novel type of palladium-catalyzed cascade cyclization coupling reaction that proceeds with suppressed beta-hydride elimination has been found. One of the N-sulfonyl oxygens is suggested to coordinatively stabilize an alkylpalladium intermediate, thus preventing the intermediate from the usual beta elimination. This is the first sequential palladium-catalyzed coupling reaction where the Suzuki and Heck reactions can compete. The reaction provides an efficient synthetic route to 4-methylene-3-arylmethylpyrrolidines, which are not readily available by other routes. PMID- 10810711 TI - Synthesis of the C22-C26 tetrahydropyran segment of phorboxazole by a stereoselective prins cyclization AB - [formula: see text] Tetrahydropyran rings are found in many complex natural products, and the segment-coupling Prins cyclization is an effective strategy for their synthesis. We report a four-step, stereoselective synthesis of the C20-C27 tetrahydropyran segment of phorboxazole. The key step is a Prins cyclization induced by catalytic BF3.OEt2. PMID- 10810712 TI - An extremely stable interwoven supramolecular bundle AB - [formula: see text] A tritopic receptor, in which three benzo[24]crown-8 macrorings are fused onto a triphenylene core, forms very strong supramolecular bundle-like complexes in CDCl3/CD3CN solution with trifurcated trications wherein three dibenzylammonium ions are linked to a central benzenoid core. The triply threaded superbundles, one of which has been characterized in the solid state by X-ray crystallography, dissociate completely via doubly and singly threaded intermediates when CD3SOCD3 is added progressively to the CDCl3/CD3CN solution. PMID- 10810713 TI - Chiral catalyst optimization using both solid-phase and liquid-phase methods in asymmetric aza Diels-Alder reactions AB - [formula: see text] In the presence of 1-5 mol % of a chiral zirconium catalyst, aza Diels-Alder reactions of aldimines with Danishefsky's dienes proceeded smoothly to afford the corresponding piperidine derivatives in high yields with high enantioselectivities. For the catalyst optimization, solid-phase and liquid phase methods were successfully used. In the solid-phase approach, polymer supported (R)-1,1'-binaphthols (BINOLs) have been synthesized and rapid optimization using the solid-phase reactions has been achieved. On the other hand, novel chiral zirconium cyanides were developed as excellent catalysts using the liquid-phase approach. PMID- 10810714 TI - Stereoselective synthesis of carbocyclic L-4'-fluoro-2',3'-dideoxyadenosine AB - [formula: see text] L-(1'S,3'S)-9-[3-Fluoro-3-(hydroxymethyl)cyclopentan-1 yl]adenine 15 has been synthesized from ester 2, which can be conveniently prepared from 2,3-isopropylidene-D-glyceraldehyde 1 in six steps. The key ring closure has been accomplished through an intramolecular nucleophilic substitution reaction. PMID- 10810715 TI - An efficient diamine.copper complex-catalyzed coupling of arylboronic acids with imidazoles AB - [formula: see text] A novel diamine.copper complex-catalyzed intermolecular coupling of arylboronic acids (1) with imidazoles (3) is described. In the presence of a catalytic amount of [Cu(OH).TMEDA]2Cl2 (2), arylboronic acids (1) react smoothly with imidazoles (3) in dichloromethane at room temperature to give a variety of N-arylimidazoles (4) in good to excellent yields. PMID- 10810716 TI - Silylcupration of allenes and subsequent electrophilic trapping by allylic phosphates. A novel approach to silylated 1,4-diene systems AB - [formula: see text] Silylcupration of allenes with a lower order silylcuprate reagent followed by an electrophilic trapping reaction with allylic diphenyl phosphates has been studied. This reaction provides a new route to 1,4-diene systems having an allylsilane moiety. PMID- 10810717 TI - Pd(0)-catalyzed cross-coupling reactions of boron derivatives with a lactam derived N-Boc enol triflate AB - [formula: see text] The cross-coupling reaction of 2-(1-alkenyl)-1,3,2 benzodioxaboroles, obtained from alkynes and catecholborane, and other boron derivatives with a lactam-derived N-Boc enol triflate occurred under very mild conditions in a THF-water medium employing (Ph3P)2PdCl2 as a catalyst, providing the corresponding 6-substituted N-Boc 3,4-dihydro-2H-pyridines in high yields. PMID- 10810718 TI - Efficient syntheses and ring-opening reactions of trans- and cis-oxazoline-5 carboxylates AB - [formula: see text] cis- and trans-Oxazoline-5-carboxylates were synthesized efficiently from isopropyl trans-cinnamate utilizing the Sharpless AA reaction. trans-Oxazoline was much more reactive than the cis-isomer toward ring opening reactions. From ab initio molecular calculations, the cis-isomer was predicted to be less reactive than the trans-isomer by 2.7 kcal/mol. Both syn and anti acetylthio esters and anti diamino esters were synthesized from these cis- and trans-oxazoline-5-carboxylates. PMID- 10810719 TI - A His-Pro-Aib peptide that exhibits an Asx-Pro-turn-like structure AB - [formula: see text] Small peptides 1 and 2, which differ only in that 1 possesses a BOC-Phe residue at the N-terminus, where 2 bears a BOC-(tau-benzyl)His, were found to exhibit very different structures. In both the solid state (X-ray) and in solution (NMR and IR), peptide 1 exists as a beta-turn, whereas peptide 2 exists in a conformation that resembles the Asx-Pro motif. PMID- 10810720 TI - Epoxide formation by ring closure of the cinnamyloxy radical AB - [formula: see text] The cinnamyloxy and oxiranyl benzyl radicals were generated by photolysis of alkyl 4-nitrobenzenesulfenates. The yet unprecedented epoxide ring formation from a primary alkoxy radical was observed. Experimental evidence supports the fact that the mode of ring opening of the oxiranyl carbinyl radical system is thermodynamically driven. B3LYP/6-31G* calculations indicate that the closed form of the radical is approximately 5 kcal/mol more stable than the open one. PMID- 10810721 TI - Rational design of chiral nanoscale adamantanoids AB - [formula: see text] Utilizing coordination as a motif, the self-organization of six ditopic and four tritopic building blocks leads to the formation of nanoscale adamantanoid frameworks. PMID- 10810722 TI - A convenient method for removing all highly-colored byproducts generated during olefin metathesis reactions AB - [formula: see text] Addition of a very modest amount of lead tetraacetate (1.50 equiv relative to the amount of Grubbs catalyst) to ring-closing metathesis reaction mixtures effectively removes all colored ruthenium and phosphine impurities to deliver colories reaction products. PMID- 10810723 TI - Tandem deployment of indium-, ruthenium-, and lead-promoted reactions. Four carbon intercalation between the carbonyl groups of open-chain and cyclic alpha diketones AB - [formula: see text] An efficient strategy for the conversion of 1,2-diketones into saturated 1,6-diketones and delta 2,3/delta 3,4-unsaturated congeners thereof is reported. PMID- 10810724 TI - Generation and trapping of isobenzofuran intermediates formed in the coupling of Fischer carbene complexes and o-alkynylbenzoyl derivatives AB - [formula: see text] The coupling of o-alkynylbenzoyl derivatives with carbene complexes has been investigated. The reaction initially affords isobenzofuran derivatives, which convert to alkylidenephthalan derivatives or can be trapped by various dienophiles to afford benzo-oxanorbornene derivatives. PMID- 10810725 TI - 7,11-cyclobotryococca-5,12,26-triene, a novel botryococcene-related hydrocarbon occurring in natural environments AB - [formula: see text] With the unambiguous structural characterization of 7,11 cyclobotryococca-5,12,26-triene 1 by NMR, a novel C30 botryococcene-related hydrocarbon skeleton bearing a midchain five-membered ring has been identified in organic-rich sediments from Lake Cadagno (Switzerland). This compound may originate from the green alga Botryococcus braunii (race B) widespread in recent and past environments. Alternatively, it may also be a yet unreported lipid of green sulfur photosynthetic bacteria (Chlorobiaceae) or a microbial transformation product of the farnesol produced by the latter. PMID- 10810726 TI - Radical cyclization of sugar-derived beta-(alkynyloxy)acrylates: synthesis of novel fused ethers AB - [formula: see text] Tributyltin radical mediated cyclization of carbohydrate derived beta-(alkynyloxy)acrylates leading to highly functionalized cis- and trans-fused bicyclic ethers of various ring sizes is described. The efficacy of the radical cyclization is nicely illustrated in the iterative construction of a trans-fused tricyclic tetrahydropyran. PMID- 10810727 TI - Synthesis of two hyaluronan trisaccharides AB - [formula: see text] The synthesis of two hyaluronan trisaccharides, methyl O (beta-D-glucopyranosyluronic acid)-(1,3)-O-(2-acetamido-2-deoxy-beta-D glucopyranosyl)-(1,4)-O-beta-D- glucopyranosiduronic acid and methyl O-(2 acetamido-2-deoxy-beta-D-glucopyranosyl)-(1,4)-O-beta- D-glycopyranosyluronic acid)-(1,3)-O-(2-acetamido-2-deoxy-beta-D-glucopyranoside, are described. Construction of the target molecules was achieved though a combination of the phenyl sulfoxide and trichloroacetimidate glycosylation methodologies. This is the first report on the synthesis of the beta-methyl derivatives, which represent the smallest fragments that incorporate all the structural features of polymeric hyaluronan. PMID- 10810728 TI - Temporary in situ aluminum and zinc tethering in Diels-Alder reactions AB - [formula: see text] Temporary tethering using aluminum or zinc in Diels-Alder reactions made possible the use of otherwise "noncompatible" combinations of dienes and dienophiles, resulting in the one-step formation of substituted cyclohexene 1,2-bis-methanols. Excellent regioselectivity and also significant stereoselectivity were obtained. PMID- 10810729 TI - Sulfur-containing palladacycles as catalyst precursors for the Heck reaction AB - [formula: see text] The air, water, and highly thermally stable sulfur-containing palladacycles, mainly derived from the ortho-palladation of benzylic thioethers, are exceptional catalyst precursors for the Heck reaction. The reaction can be performed with aryl lodides, bromides, and chlorides, with acrylic esters and styrene, leading to turnover numbers up to 1,850,000. PMID- 10810730 TI - Aldol reaction under solvent-free conditions: highly stereoselective synthesis of 1,3-amino alcohols AB - [formula: see text] A method for the highly stereoselective synthesis of 1,3 amino alcohols based on the indium trichloride-catalyzed Mukaiyama aldol reaction of keto ester (R,S)-1 under solvent-free conditions has been developed. The high selectivity observed can be explained on the basis of the shielding of one face by a remote substituent. PMID- 10810731 TI - Direct amination of olefins through sequential triazolinedione ene reaction and carbanion-assisted cleavage of the N-N urazole bond AB - [formula: see text] Allylic amines 5 are obtained in 30-55% overall yields by the base-catalyzed hydrolysis of trialkylated allylic urazoles 3; the latter are prepared by the TAD ene reaction of the appropriate olefin and further N alkylation with alpha-bromoacetophenone. The proposed mechanism for this novel urazole rupture is based on the generation of a carbanion adjacent to the hydrazide functionality, which induces urazole ring-opening by cleavage of the N N bond. PMID- 10810732 TI - [Bis] AB - [formula: see text] The generation of (2-PyMe2Si)2CHLi was easily accomplished by the deprotonation of (2-PyMe2Si)2CH2 using n-BuLi in Et2O. Thus generated (2 PyMe2Si)2CHLi was found to react with a variety of aldehydes and ketones to give the corresponding vinylsilanes in extremely high yields with complete stereoselectivities. PMID- 10810733 TI - Iron-catalyzed Doyle-Kirmse reaction of allyl sulfides with (trimethylsilyl)diazomethane AB - [formula: see text] Iron salts efficiently catalyze the Doyle-Kirmse reaction of allyl sulfides with (trimethylsilyl)diazomethane and ethyl diazoacetate in dichloroethane at 83 degrees C. Competitive dimerization is less of a problem with (trimethylsilyl)diazomethane than with ethyl diazoacetate. Good results are obtained using only 1.5 equiv of (trimethylsilyl)diazomethane, even without slow addition. Phosphine ligands affect the kinetics, but not the diastereoselectivity. Dppe and BINAP lead to higher yields than dppp, but no enantioselection was detected with R-(+)-BINAP. PMID- 10810734 TI - Mechanistic studies of the tRNA-modifying enzyme QueA: a chemical imperative for the use of AdoMet as a "ribosyl" donor. AB - [formula: see text] The enzyme S-adenosylmethionine:tRNA ribosyltransferase isomerase (QueA) catalyzes the penultimate step in the biosynthesis of the tRNA nucleoside queuosine, a unique ribosyl transfer from the cofactor S adenosylmethionine (AdoMet) to a modified-tRNA precursor. The use of AdoMet in this way is fundamentally new to the chemistry of this important biological cofactor. We report here the first mechanistic studies of this remarkable enzyme, and we propose a chemical mechanism for the reaction consistent with our experimental observations. PMID- 10810735 TI - Stealth star polymers: a new high-loading scaffold for liquid-phase organic synthesis AB - [formula: see text] Polyethylene glycol (PEG) has proven to be a versatile soluble-polymer support for organic synthesis, though the use of PEG has been limited by its relatively low loading (0.5 mmol/g or less). We have developed a new high-loading (1 mmol/g) soluble-star polymer based on a cyclotriphosphazene core with PEG arms that exhibit superior precipitation properties compared with those of linear PEG. Additionally, the heterocyclic core does not add interfering signals to the 1H or 13C NMR. PMID- 10810736 TI - Nuclear magnetic resonance studies of helical dipyrromethene-zinc complexes AB - [formula: see text] Analysis of helical chirality within dinuclear dipyrromethene Zn(II) complexes has been achieved with the use of 1H NMR spectroscopy. The use of AgFOD and chiral lanthanide shift reagents gives fully resolved resonances attributable to two diastereomeric helical complexes. PMID- 10810737 TI - Asymmetric desymmetrization of 2,2',6,6'-tetrahydroxybiphenyl through annulation with enantiomerically pure bis(mesylate) AB - [formula: see text] 2,2',6,6'-Tetrahydroxybiphenyl undergoes a facile annulation reaction with bis(mesylate) derived from (S)-1,2-propanediol in the presence of Cs2CO3 to give the corresponding asymmetric desymmetrization product of S axial chirality with exclusive diastereoselectivity. The desymmetrization product can be utilized as a versatile chiral building block in asymmetric synthesis of axially chiral 6,6'-disubstituted 2,2'-biphenyldiols. PMID- 10810738 TI - Chemistry of beta-functionalized alpha-nitroso ethylenes. Methyl beta-nitroso acrylate as heterodienophile in AB - [formula: see text] beta-Functionalized nitroso alkene 2, obtained from methyl beta-nitropropionate 1 and N,O-bis(trimethylsilyl)acetamide, can function as a good heterodienophile in Diels-Alder reactions. Therefore, 2 was trapped by cyclic dienes to give adducts 4 with the corresponding stereoselectivity. Cycloadduct 4a undergoes retro-[4 + 2]-cycloaddition at 33 degrees C in solution; thus 4a can be used to generate nitroso alkene 2 in neutral medium. Cyclopentadiene reacts with adduct 4a according to an endo-(4 + 2]-cycloaddition scheme to give cycloadduct 5 in low yield. PMID- 10810739 TI - Thiopyran route to polypropionates: aldol diastereoselectivity of linear and two directional iterative homologations AB - [formula: see text] Aldol reaction of tetrahydro-4H-thiopyran-4-one with racemic 1,4-dioxa-8-thiaspiro[4.5]decane-6-carboxaldehyde is easily controlled to give the 2,3-anti-3,4-syn or the 2,3-syn-3,4-syn adduct. Aldol homologations of these beta-hydroxy ketones with the same aldehyde occur with considerable mutual kinetic enantioselection (MKE) and, in each case, selectively give one of the eight possible diastereomers. Similar reactions of related beta-methoxy ketones are also very diastereoselective but proceed without significant MKE, resulting in two diastereomers. The adducts can be used for polypropionate synthesis. PMID- 10810740 TI - Selective reduction of 2- AB - [formula: see text] NaBH4 reduction of 2,5-bis[(4'-(n alkoxy)benzoyl)oxy]benzaldehydes produces primarily the rearranged phenol, which does not generate liquid crystalline phases when laterally attached to a polymer backbone. Rearrangement is prevented by quenching the intermediate benzyl alkoxide with a weak acid. For example, 2,5-bis[(4' (methoxy)benzoyl)oxy]benzaldehyde is selectively reduced to 2,5-bis[(4' (methoxy)benzoyl)oxy]-benzyl alcohol with less than 5% intramolecular transesterification using 2-3 equiv of NaBH4 in the presence of 20-30 equiv of acetic acid (1:10 NaBH4/AcOH). PMID- 10810741 TI - Synthesis of enamides related to the salicylate antitumor macrolides using copper mediated vinylic substitution AB - [formula: see text] A new approach to the assembly of enamides is described using copper(I) carboxylate-catalyzed substitution of vinyl iodides and amides. Modified reaction conditions have been developed to synthesize the O-methyloxime enamide side chains related to the natural products lobatamides A-F, oximidine I and II, and CJ-12,950. PMID- 10810742 TI - Enantioselective synthesis of 2-substituted cyclobutanones AB - [formula: see text] An enantioselective synthesis of 2-substituted cyclobutanones has been achieved by sequential application of the titanium-mediated cyclopropanation of alpha-hydroxy esters and the pinacol-type rearrangement of the resulting alpha-hydroxycyclopropylcarbinols. PMID- 10810743 TI - Recognition of the nonpolar base 4-methylindole in DNA by the DNA repair adenine glycosylase MutY. AB - [formula: see text] The DNA repair adenine glycosylase MutY efficiently recognizes 7-deaza-2'-deoxyadenosine (Z) and its nonpolar isostere 4-methylindole beta-deoxynucleoside (M) opposite 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG) and G in DNA. Both wild-type and truncated MutY exhibit a 10- to 20-fold higher affinity for a duplex containing OG:M than OG:Z. More efficient recognition of M over Z by MutY may be to due the lack of hydrogen bonding with the OG that facilitates nucleotide flipping during the substrate recognition process. PMID- 10810744 TI - Granulation of acetaminophen by a rotating fluidized-bed granulator. AB - The purpose of this research was to evaluate the use of a rotating fluidized-bed granulator to produce acetaminophen granules with sufficient binding force between particles and good plasticity in tablets. Ethenzamide and ascorbic acid were used to compare the relationship between granulation and the sample wetness. It was revealed that a blade rotation rate of 300 rpm, inlet air flow rate of 42 m3/hr, and spraying pressure of 1.5 kg/cm3 produced tablets with the best properties. The granule and tablet properties of ethenzamide and ascorbic acid were compared to those of acetaminophen. These compounds showed different wetting behaviors with water and different compression behaviors. With an increase in medicament content, tablet hardness increased except for the ascorbic acid formulation. Capping and sticking were observed in acetaminophen and in ascorbic acid, respectively, and acetaminophen and ethenzamide showed prolonged disintegration time. PMID- 10810745 TI - Moisture uptake and its influence on pressurized metered-dose inhalers. AB - The objective of this study was to investigate moisture ingress into pressurized metered dose inhalers (pMDIs) containing hydrofluoroalkane (HFA) propellants and the consequences of this ingress. Moisture ingress into the pMDIs containing tetrafluoroethane (HFA 134a) or heptafluoropropane (HFA 227) was evaluated and modeled. The influence of water level in pMDIs on the stability of pMDIs containing triamicinolone acetonide (TAA) and beclomethasone dipropionate (BDP) in terms of particle growth, fine particle fraction, and drug solubility in the propellant system was evaluated using scanning electron microscopy, particle size analysis, single-stage impaction, and HPLC. The water level in HFA-containing pMDIs increased during storage and the process obeyed a diffusion model. HFA 134a had a greater tendency to take up moisture from the environment than did HFA 227. Unlike TAA, the propensity for particle growth of the suspended BDP in HFA propellants was significantly depressed by the increase in water level in the pMDIs. As a result, the fine particle fraction of the emitted BDP aerosols significantly increased as the water level in the HFA propellant was increased. Moisture ingress into pMDIs containing HFAs occurred during storage. The influence of the increased water level in pMDIs on the physical stability of the pMDI formulation and the dose delivery performance was a function of the composition of the internal lining of the container, the type of drug and propellant, and storage temperature. PMID- 10810746 TI - Disintegration propensity of tablets evaluated by means of disintegrating force kinetics. AB - The aim of this work was to measure the disintegrating force concomitantly with tablet disintegration, in order to evaluate the disintegration propensity of tablets. Disintegration and dissolution were measured on tablets containing two poorly soluble drugs (diclofenac sodium or ketoprofen), including different percentages of two disintegrants (Explotab or Ac-Di-Sol). Because of the experimental setup, the disintegrating force measured was the result of the force generated by disintegrant swelling and dissipated by tablet disintegration. The disintegrating force versus time curves had shapes ranging from a skewed distribution curve to a bell-shaped curve, depending on slow or rapid disintegration of tablets, respectively. Interestingly, the shape of the resulting curves was very sensitive to the composition of the tablet. When the disintegrant in the formula was increased, the force-time curve approached the bell-like shape. The disintegration propensity of the tablet can be evaluated by the disintegrating force development during disintegration. The disintegration improvement of the formula can be predicted. The disintegrating force curve allows for the clear identification of the optimal percentage of disintegrant to be used. PMID- 10810747 TI - Content-based image retrieval: a new promising technique in powder technology. AB - The aim of the present study was to introduce a new technique for analyzing powders by examining the content information of images of pharmaceutical powder systems. Texture features of images of microcrystalline cellulose were compared by using a content-based image retrieval system (CBIR), QBIC (Query-by-Image Content). The rank order and image similarities were compared to particle sizes and appearances of different mixtures. The image order of the similarity values was in close agreement with the appearance and particle size of the mixtures. When the image of pure Avicel PH 101 was used as a query image, the most similar images were always from images of mixtures with a large number of particles with smaller particle mean sizes. When images of pure Avicel PH 200 were used as a query image, the closest matches of image similarity were from images of mixtures with a larger amount of larger particles. The results show that the CBIR system extracts applicable content information on images of powders, but the texture features used were not totally adequate for analysis of the powders used. In general, content-based image retrieval seems to be a promising approach to efficiently use the vast image information that is available from pharmaceutical powders. Nevertheless, to achieve an efficient CBIR tool for powder technology requires development of substantial algorithms for feature extraction. PMID- 10810748 TI - Mercury porosimetry of mannitol tablets: effect of scanning speed and moisture. AB - Purpose of the work was to study the effect of the scanning speed of mercury porosimetry and moisture content of the sample on the mercury porosimetry result for mannitol tablets. Tablets were compressed at three different compression pressures from nonhygroscopic mannitol powder and granules. Pore structure of tablets was determined with three different scanning speeds of a high-pressure mercury porosimeter after storage in three different moisture conditions. With low scanning speed, smallest pores of tablets were determined more accurately. Small amounts of moisture, even as low as 1%, before evacuation in nonhygroscopic mannitol tablets decrease the porosity. Decrease in porosity was observed at a pore diameter range of 50-1000 nm, not at the smallest determined pores. Thus, the role of water in pharmaceutical samples appears to be complicated. Reasonably slow scanning is recommended in high-pressure mercury porosimetry. If total pore volume is the only parameter of interest, fast scanning can be used. Pretreatment of the samples by proper drying before mercury porosimetry is important. PMID- 10810749 TI - The effects of pH and PEG 400-water cosolvents on oxytetracycline-magnesium complex formation and stability. AB - The effects of pH and PEG 400 on the stoichiometry, conformation, and stability of the magnesium-oxytetracycline (Mg+2-OTC) complex were evaluated. Circular dichroism (CD) and HPLC were used to investigate Mg+2-OTC complex formation and determine the stability of the complexes formed. The stoichiometry of the complex was determined to be a 1:1 molar ratio of Mg+2 to OTC regardless of changes in pH, in the range 7-10, and regardless of the percentage of polyethylene glycol (PEG) 400 in solution. CD showed that the conformation assumed by Mg+2-OTC complex is sensitive to changes in pH, however, little to no effect was found when the PEG 400 concentration was varied. PEG 400 was found to effect the magnitude of complexation as evident by the dependence of CD peak intensity on the cosolvent concentration in solution. The Job's method confirmed that the formation of this complex increased with increasing PEG 400 concentration and was most favored at pH 8. HPLC analyses of OTC solutions at pH 9 revealed the formation of multiple degradation products after storage at 50 degrees C. The incidence and magnitude of OTC degradation products were reduced in the presence of Mg+2 and PEG 400. Despite the HPLC results of maintained OTC stability in magnesium-complexed solutions over time, visual inspection showed these solutions to have darkened, indicating that an oxidative process is responsible for initial degradation of OTC. Therefore, the need for additional measures (i.e., antioxidants) was established to ensure the long-term stability of OTC in solution. PMID- 10810750 TI - Comparison of various injectable protein-loaded biodegradable poly(lactide-co glycolide) (PLGA) devices: in-situ-formed implant versus in-situ-formed microspheres versus isolated microspheres. AB - The purpose of this research was to prepare various injectable, protein (cytochrome c)-loaded biodegradable poly(lactide-co-glycolide) (PLGA) devices by a novel microencapsulation method and to compare their characteristics. Syringeable mixtures of polymer and protein solidified upon injection when coming in contact with water, and formed a solid matrix-type implant or microspheres (in situ-formed implant or in-situ-formed microspheres, respectively) with cytochrome c entrapped. These devices exhibited different characteristics in terms of in vitro cytochrome c release profile, percentage cytochrome c encapsulation efficiency, and particle size. The burst effect from these devices exhibited the following trend: in-situ-formed implant > in-situ-formed microspheres > isolated microspheres. The in-situ-formed microspheres were larger in size than the isolated microspheres. Also, the isolated microspheres exhibited the slowest release of cytochrome c, whereas the in-situ-formed implant exhibited the fastest release. The microencapsulation process can produce various drug-loaded injectable biodegradable PLGA devices having different characteristics. PMID- 10810751 TI - Use of the near-infrared reflectance method for measurement of moisture content during granulation. AB - The purpose of this study was to demonstrate the use of a near-infrared (NIR) method for in-process control of a placebo formulation. An NIR setup with a multichannel detector was applied in the measurement of water during fluidized bed granulation. The effects of two critical granulation parameters were studied using the central composite design. The present NIR setup with three wavelengths proved applicable for in-line moisture measurement. The 1990 nm signal was used for measurement of water and the 1745 and 2145 nm signals were used to correct the change in spectra baseline during granulation. Variations in inlet air conditions proved to be critical factors, explaining differences in the granule size distributions. Differences in granule moistening and drying rates resulting from varying inlet air conditions could be measured with the NIR setup. The moisture content of granules at the end of the spraying phase explained part of the differences in granule size distributions. The moisture content of granules at the end of the drying phase affected the tableting behavior of granules. The results suggested that direct measurement of granule moisture content facilitates the in-process control of the granulation. PMID- 10810752 TI - Lecithin inverse microemulsions for the pulmonary delivery of polar compounds utilizing dimethylether and propane as propellants. AB - Lecithin inverse microemulsions were investigated as a means of pulmonary drug delivery, utilizing dimethylethyleneglycol (DMEG) and hexane as models for dimethyl ether (DME) and propane, respectively. Addition of lecithin to the model propellant mixtures increased the solubility of water in a nonlinear, solvent dependent manner. The concentration of water necessary to fully hydrate cobalt(II) decreased as the solvent composition was varied from DMEG to hexane. Water proton chemical shift increased in the presence of lecithin, with the largest increases in high hexane content samples. Equilibrium dialysis and component diffusion rate determination (by pulsed-field gradient [PFG]-NMR) indicated the quantity of water associated with the dispersed phase. Collectively, these methods demonstrated that a greater fraction of water was associated with the microemulsion-dispersed phase as the solvent was varied from DMEG to hexane. Iodine solubilization indicated microemulsion formation (operational critical micelle concentration [cmc], 10 moles water per mole lecithin) at approximately 10(-4)-10(-5) molal lecithin. NMR data (trimethylammonium proton chemical shift, water, and lecithin T1) were consistent with microemulsion formation. Water-soluble compounds dissolved in lecithin inverse microemulsions in a lecithin- and water-dependent manner. Experiments with DME/lecithin demonstrated microemulsion characteristics similar to those in the model propellant. DME/lecithin metered-dose inhalers (MDIs) produced a particle size and a fine particle fraction (36% by twin impinger method) suitable for pulmonary drug delivery. PMID- 10810753 TI - Quantifying crystalline form composition in binary powder mixtures using near infrared reflectance spectroscopy. AB - The objectives of this study were to assess the utility of near-infrared reflectance spectroscopy (NIRS) in differentiating crystalline forms of pharmaceutical materials and determine the accuracy of this technique in quantifying crystalline forms of solids in binary mixtures. Various crystalline forms of sulfamethoxazole, sulfathiazole, lactose, and ampicillin, independently characterized with other methods, were analyzed qualitatively and quantitatively. The observed differences in near-infrared (NIR) spectra of crystalline form pairs were interpretable on the basis of the features of their crystalline and molecular structures and mid-infrared spectra. NIR spectra of binary physical mixtures of crystalline form pairs were obtained directly through glass vials over the wavelength range of 1100-2500 nm. The calibration lines were constructed using an inverted least-squares regression method. The ratio of the response of the second derivative of the reflectance spectra at two wavelengths was plotted versus crystal form composition. The correlation coefficients for plots of predicted versus theoretical composition were generally greater than 0.99 and standard errors were all low. Parallel studies comparing the NIRS method to a quantitative x-ray powder diffraction method using sulfamethoxazole and sulfathiazole confirmed the accuracy of the results. Additional NIRS studies were conducted in the 0-10% composition range with ampicillin and sulfamethoxazole. These results indicated that prediction down to the 1% level was possible. This study demonstrates that NIRS can be used as a quantitative physical characterization method, is comparable in accuracy to other techniques, and is capable of detecting low levels of one crystal form in the presence of another. PMID- 10810754 TI - Direct pelletization in a rotary processor controlled by torque measurements. I. Influence of process variables. AB - The aim of this study was to elucidate the feasibility of using torque measurement to control the end point of a wet pelletization process in a rotary processor at varying levels of friction plate rotation speed, air gap pressure difference, and batch size. A 1:1 mixture of lactose monohydrate (200 mesh) and microcrystalline cellulose (PH-101) was granulated into pellets in an instrumented laboratory scale rotary processor using water as aqueous binder liquid. A full factorial designed study was performed to investigate the influence of the friction plate rotation speed (600 and 1200 rpm), the air gap pressure difference (1 and 3 kPa), the torque increase (0.4 and 0.8 N.m) and the batch size (500 and 1000 g) on the pellet properties. All pellets produced were round and showed a narrow size distribution. The geometric mean diameter varied from 400 to 1900 microns with a good reproducibility. Increasing the batch size and the rotation speed led to smaller pellets, whereas a higher torque produced larger pellets. This study showed that the process can be controlled by means of the torque increase because it was possible to produce pellets of a reproducible size by stopping the liquid addition at a certain torque level. PMID- 10810755 TI - In situ dehydration of carbamazepine dihydrate: a novel technique to prepare amorphous anhydrous carbamazepine. AB - The purposes of this project were to prepare amorphous carbamazepine by dehydration of crystalline carbamazepine dihydrate, and to study the kinetics of crystallization of the prepared amorphous phase. Amorphous carbamazepine was formed and characterized in situ in the sample chamber of a differential scanning calorimeter (DSC), a thermogravimetric analyzer (TGA), and a variable temperature x-ray powder diffractometer (VTXRD). It has a glass transition temperature of 56 degrees C and it is a relatively strong glass with a strength parameter of 37. The kinetics of its crystallization were followed by isothermal XRD, under a controlled water vapor pressure of 23 Torr. The crystallization kinetics are best described by the three-dimensional nuclear growth model with rate constants of 0.014, 0.021, and 0.032 min-1 at 45, 50, and 55 degrees C, respectively. When the Arrhenius equation was used, the activation energy of crystallization was calculated to be 74 kJ/mol in the presence of water vapor (23 Torr). On the basis of the Kissinger plot, the activation energy of crystallization in the absence of water vapor (0 Torr water vapor pressure) was determined to be 157 kJ/mol. Dehydration of the dihydrate is a novel method to prepare amorphous carbamazepine; in comparison with other methods, it is a relatively gentle and effective technique. PMID- 10810756 TI - Production of Eudragit microparticles by spray-drying technique: influence of experimental parameters on morphological and dimensional characteristics. AB - The aim of this study was to evaluate the influence of operating parameters on the characteristics of methacrylate microparticles prepared by spray-drying technique. Eudragit microparticles were prepared by a spray-drying method. The influence of different experimental parameters (i.e., solvent, feed rate, air flow rate, air-drying temperature, and aspiration flow rate) on microparticle morphology, size distribution, and recovery was studied. In addition, different Eudragit types and Eudragit RS concentrations were employed. Optical and electron microscopy were employed to analyze microparticle morphology and dimensional distribution. Finally, prednisolone as model drug was encapsulated in Eudragit RS microparticles. Low feed rate values yielded the best results in terms of microparticle morphology. Changes in nebulizing air flow did not result in a corresponding effect on microparticle characteristics. An increase of air-drying temperature resulted in a reduction of microparticle dimension and recovery. A low flow of drying air was preferable because this resulted in microparticles with an optimal morphology. The polymer concentration affected both morphology and dimensions of microparticles. The encapsulation of prednisolone led to good incorporation efficiencies without altering percentage of recovery, morphology, and mean dimension of the microparticles. The selection of appropriate parameters yielded spray-dried Eudragit RS microparticles characterized by good morphology and narrow dimensional distribution. PMID- 10810757 TI - Kinetics of a model nucleoside (guanosine) release from biodegradable poly(DL lactide-co-glycolide) microspheres: a delivery system for long-term intraocular delivery. AB - The objective of this study was to prepare poly(DL-lactide-co-glycolide) (PLGA) microspheres containing guanosine as a model drug for intraocular administration. Microspheres were prepared by solvent evaporation technique using o/w emulsion system. The influence of composition and molecular weight of PLGA, drug loading efficiency, microsphere size, and in vitro and in vivo release rates were determined. Differential scanning calorimetry (DSC) and FTIR studies were conducted to examine the guanosine-polymer interaction. In vitro release studies indicated that the permeant release from microspheres exhibits an initial burst followed by slow first-order kinetics. Ascending molecular weights of the polymers generated progressively slower release rates. Three different sizes of microspheres were prepared. The release continued for 7 days with a maximum of 70% of the content released within that time period. DSC and FTIR studies showed no polymer-guanosine interaction. A novel microdialysis technique was used to examine the initial release kinetics from microspheres in isolated vitreous humor. This technique was also employed to observe in vivo intravitreal release in albino rabbits. A good correlation exists between in vitro and in vivo release rates from both 75 and 140 kDa PLGA microspheres. Guanosine-loaded microspheres could be prepared for once-a-week intravitreal injection with minimum required concentration maintained throughout the dosing interval. Because the structural and solubility characteristics of guanosine are similar to those of acyclovir and ganciclovir (two acycloguanosine analogues effective against herpes simplex virus [HSV-1] and cytomegalovirus [CMV], respectively), similar biodegradable polymer based microsphere technology can be employed for the long-term intraocular delivery of these two drugs. PMID- 10810758 TI - Enhancing initial release of peptide from poly(d,l-lactide-co-glycolide) (PLGA) microspheres by addition of a porosigen and increasing drug load. AB - The objective of this study was to evaluate formulation variables such as drug load and addition of a porosigen in achieving an increased initial release of peptide from poly(d,l-lactide-co-glycolide) (PLGA) microspheres by altering carrier characteristics. Leuprolide acetate-loaded PLGA microspheres were prepared by a solvent-extraction-evaporation process and were characterized for their drug load (HPLC assay), bulk density (tapping method), size distribution (dynamic light scattering), specific surface area (Brunauer-Emmett-Teller [BET] analysis), surface morphology (scanning electron microscopy), in vitro drug release (at 37 degrees C), and in vivo efficacy (suppression of rat serum testosterone). Increasing the drug load, and adding various amounts of calcium chloride to organic and aqueous phases of the emulsion during processing yielded particles with increased porosity, lower bulk density, higher specific surface area, and accordingly higher initial release. In an animal model, these formulations showed a faster onset of testosterone suppression compared to microspheres without higher drug load or calcium chloride. The approaches employed in this study were found to be effective in avoiding the therapeutic lag phase usually observed with microencapsulated macromolecular drugs. PMID- 10810759 TI - Formulation design studies of atenolol tablets. PMID- 10810760 TI - Muscle injury and repair. AB - Muscle injury triggers a sequence of events that begin with a host inflammatory response that is followed by muscle fiber regeneration and new collagen synthesis. The inflammatory response involves at least three types of cells, including neutrophils, ED1+ macrophages, and ED2+ macrophages. A host of growth factors and cytokines appear to play a role in the inflammatory process and repair of the damaged tissue. Satellite cells play an integral role in normal development of skeletal muscle by providing a source for postmitotic myonuclei. In addition, the satellite cell is essential to the repair of injured muscle by serving as a source of myoblasts for fiber regeneration. At the same time muscle fiber regeneration is occurring, there is expression of types I and III collagen that under certain circumstances can lead to scarring and fibrosis. Current studies of treatment of muscle injury often incorporate investigations of basic principles of injury and repair together with clinical experience and principles in an effort to coordinate basic science and outcome studies. PMID- 10810761 TI - Injury and repair of tendons and ligaments. AB - Tendons and ligaments are fibrous connective tissues that facilitate stability and motion of joints. Significant dysfunction and disability may result from suboptimal healing of tendon and ligament injuries. Extensive research continues to further understand the complex healing pathways that are involved when these structures are damaged. The combination of advances in tissue engineering, surgery, and rehabilitation will provide new pathways of improving tendon and ligament healing. PMID- 10810762 TI - Cartilage injury and repair. AB - Articular cartilage is a complex structure that, once damaged, has little capacity for permanent repair. The problem lies in the inability of the body to regenerate tissue with the appropriate macromolecular constituents and architecture of normal hyaline cartilage. Although full-thickness defects are capable of stimulating a repair response, the resulting fibrocartilage is inferior and cannot withstand long-term, repetitive use. Numerous surgical approaches that involve penetration of subchondral bone offer short-term to moderate-term relief of symptoms, whereas other approaches have seen significant improvement through transplantation of osteochondral and periosteal tissue and implantation of autologous chondrocytes. Despite these procedures, articular cartilage damage continues to be an unmet clinical problem. Improvements in biochemical and molecular biologic techniques may allow advances in the understanding of chondrocyte and cartilage biology and may provide innovative and novel approaches to stimulating the repair of articular cartilage through biologic means. PMID- 10810763 TI - Biomechanics and motion analysis applied to sports. AB - The development of motion analysis and the application of biomechanical analysis techniques to sports has paralleled the exponential growth of computational and videographic technology. Technological developments have provided for advances in the investigation of the human body and the action of the human body during sports believed to be unobtainable a few years ago. Technological advancements have brought biomechanical applications into a wide range of fields from orthopedics to entertainment. An area that has made tremendous gains using biomechanics is sports science. Coaches, therapists, and physicians are using biomechanics to improve performance, rehabilitation, and the prevention of sports related injuries. Functional analyses of athletic movements that were impossible a few years ago are available and used today. With new advancements, the possibilities for investigating the way a human interacts and reacts to environmental conditions are ever expanding. PMID- 10810764 TI - Proprioception. AB - Although definitions of proprioception may vary, its importance in preventing and rehabilitating athletic injuries remains constant. Proprioception plays a significant role in the afferent-efferent neuromuscular control arc. This control arc is disrupted with joint and soft tissue injury. Restoring proprioception after injury allows the body to maintain stability and orientation during static and dynamic activities. By focusing on aspects of neuromuscular function, such as dynamic joint stability, practitioners can design and study interventions to maximize sport and daily life neuromuscular function. Further research is necessary to elucidate how proprioceptive deficits can be remedied or compensated to improve function and prevent reinjury. PMID- 10810765 TI - Training for muscular power. AB - In sport and everyday activities, the most important attribute of skeletal muscle is the ability to generate power, a product of strength and speed of movement. Many factors influence the muscle's ability to generate power. Training for muscular power requires special care in developing the proper exercise prescription. The need for muscular power runs across a spectrum of people from elite athletes attempting to optimize sports performance to the frail elderly trying to perform simple tasks. Power development is paramount to optimal neuromuscular function. PMID- 10810766 TI - Closed kinetic chain rehabilitation for sports injuries. AB - Closed chain techniques can increase the effectiveness of rehabilitation protocols because they allow more normal physiologic activations and biomechanical motions, especially in the early rehabilitation phase. They have been shown to be effective in knee/leg rehabilitation, but are also useful in shoulder/scapula rehabilitation. Facilitation patterns allow muscles to be activated in normal sequences, but also allow them to be isolated to recover their normal strength. Closed chain exercises are an integral part of accelerated rehabilitation programs. PMID- 10810767 TI - Muscle fatigue and muscle injury. AB - The purpose of this review is to acquaint the reader with the neurobiology of muscle fatigue. Muscle fatigue is a complex, multifactorial process. The authors have covered the chain of events bringing about skeletal muscle contraction and the manner in which fatigue may affect each step. Advances in technology continue to increase understanding of central fatigue. Many excellent studies of peripheral fatigue have been designed to delineate the mechanisms that influence the excitation-contraction coupling, energy supply, and force generation processes. Although much of this work has considered mechanisms in isolation, different mechanisms may be responsible under different conditions. Fatigue is a common complaint among patients with a variety of neuromuscular and metabolic diseases. Armed with an enhanced knowledge of the mechanisms of muscle fatigue, one can more fully recognize the signs and symptoms of metabolic disorders and neuromuscular diseases and use diagnostic testing. The clinician should anticipate the role of muscle fatigue in injury and focus on injury prevention strategies, especially during the restorative phase of rehabilitation. As a clinician-scientist concerned with optimizing patients' and athletes' performance, one must design ways to identify, measure, and treat muscle fatigue. Beyond illustrating what is currently known about muscle fatigue, the authors hope this review inspires the reader to solve problems of great clinical importance to patients and athletes alike. PMID- 10810768 TI - Potential applications of gene therapy in sports medicine. AB - Gene therapy has the potential to improve greatly the management of sports medicine injuries. By the expeditious introduction of the appropriate genes to sites of injury, it should prove possible to initiate healing responses in those tissues that normally fail to heal, to accelerate healing in tissues which do naturally heal, and to improve the quality of the repair tissue. In addition, genes with anti-inflammatory properties offer novel possibilities for the control of chronic, sports related inflammation. None of these applications of gene therapy in sports medicine is yet in the clinic. It would seem, however, only a matter of time before the first clinical trials emerge. Their acceptance will be facilitated by the two human protocols that are presently underway for the treatment of rheumatoid arthritis. The present status of sports medicine gene therapy is summarized in Table 3. PMID- 10810769 TI - Computer modeling and simulation of human movement. Applications in sport and rehabilitation. AB - Computer modeling and simulation of human movement plays an increasingly important role in sport and rehabilitation, with applications ranging from sport equipment design to understanding pathologic gait. The complex dynamic interactions within the musculoskeletal and neuromuscular systems make analyzing human movement with existing experimental techniques difficult but computer modeling and simulation allows for the identification of these complex interactions and causal relationships between input and output variables. This article provides an overview of computer modeling and simulation and presents an example application in the field of rehabilitation. PMID- 10810770 TI - Imaging of sports injuries. AB - Imaging sports injuries is challenging, interesting, and evolving. The use of imaging and its various modalities depends on the clinicians' and radiologists' comfort and facility with those modalities, with the clinician playing the pivotal role. Imaging can be used to make diagnoses, define prognoses, influence treatment plans, or to preoperatively assess the severity of known pathology or look for additional injuries. But, because of the prevalence of abnormal findings in the asymptomatic population, it must be the clinician who applies the imaging findings to the specific situation. The clinician also must decide on the need for imaging; no imaging should be undertaken if it does not affect the clinical decision making. In sports injuries, just as in other parts of medicine, prevention is better than cure. When an injury does occur, however, imaging can help to define it and to guide appropriate therapy. PMID- 10810771 TI - Communicative ability in an audiological perspective. Theory and application to post-secondary school students. AB - The underlying assumption in the present study is that the individual's speech and hearing communicative ability is composed of three components, each corresponding to different functional systems of the brain: afferent functions (A) represent the auditory activity and sound perception largely corresponding to activity in the ascending auditory pathways. The central functions (C) include cortical auditory and language abilities controlled in parts of the left temporal lobe and subcortical centres. The efferent functions (E) consist of speech motor processes and articulation. A test battery of 20 tests measuring several aspects of afferent, central and efferent functions was applied to 11 hearing-impaired post-secondary school students and several control groups. All data were normalized with the normal materials as references. Individual communicative profiles were obtained from these primary data, which consisted of audiometric tests (tone and speech audiometry, impedance tests, brainstem response audiometry and phase audiometry), sound environmental tests with hearing aids (directional speech-in-noise, word localization, sound environment identification test), and language tests (reading tests, prosody, auditory memory and recall, phonology and articulation). Since the central functions cannot truly and directly be determined in hearing-imparied subjects, they were assessed under optimal listening conditions. Furthermore, central functions were estimated according to three different models: distributive, parallel model (model 1), multiplicative, serial model (model 2) and compensatory model (model 3). On the basis of these models, a three-component description of the communicative ability consisting of A,C and E functions was obtained. It was found that C and E functions were largely independent of the adult afferent functions, but C functions were negatively correctly to hearing in childhood. A preliminary comparison between the tests and a comparison between the models was performed by predicting benefit of hearing aid. Model 3 gave the best prediction. Beyond the three-component A,C, and E characterization of the students, a total communicative ability score could be calculated giving values from 37% to 79% of the normal mean. On the basis of the conceptual and statistical analyses, the test battery could be reduced to include tone 0-12 years, tone adults, word localization test (afferent); word chain, lecture test (central), articulatory test (efferent) and audiovisual test. The simple algorithm of adding the normalized loss of afferent (peripheral) function to the normalized results of the acoustic central tests seems to be promising for isolation for the central auditory capacity even in cases with peripheral impairment. It is concluded that a wider perspective is desirable in the diagnostic evaluation of the hearing-impaired individual in order to understand his communicative abilities and form a cornerstone in the planning of rehabilitation in conjunction with social and psychological factors. PMID- 10810772 TI - Endocarditis: epidemiology, diagnosis and treatment. AB - The current incidence of infective endocarditis (IE) is estimated as seven cases per 100,000 population per year and continues to increase. The prognosis is significantly influenced by proper diagnosis and adequate therapy. In cases with unconfirmed IE, besides careful clinical examination, transesophageal echocardiography is the imaging technique of choice. Culture-negative endocarditis requires either termination of antimicrobial treatment initiated without microbiological test results and reevaluation of blood samples or serological/molecular biological techniques to identify the causative organism. Antimicrobial therapy should established only after quantitative sensitivity tests of antibiotics, including evaluation of the minimal bactericidal concentrations (MBC) of established combinations of antibiotics. Concomitant kidney involvement, a significant impairment of the hepatic function or the combination therapy with oto- and/or nephrotoxic antibiotics, requires drug monitoring. Large (> 10 mm) vegetations attached to the mitral valve are linked with a high incidence of thromboembolic complications. In most of these cases, the indication for surgical intervention is given, especially if the MBC of the optimal combination of antibiotics is high (> 2 micrograms/ml). In the first three weeks after primary manifestation of signs and symptoms of IE, an index embolism is frequently followed by recurrences. If vegetations can still be demonstrated by echocardiography after an embolic event, surgical intervention should seriously be considered. Cerebral embolic events are no contraindication for surgery with use of the heart-lung machine, as long as cerebral bleeding has been excluded by cranial computed tomography immediately preoperatively and the operation is performed before a significant disturbance of the blood-brain barrier (< 72 hours) has manifested. A significant prognostic improvement has also been demonstrated for patients with early surgical intervention, after their clinical course was complicated by myocardial failure due to acute valve incompetence, acute renal failure, mitral kissing vegetations in primary aortic valve IE, and in patients with sepsis persisting for more than 48 hours despite adequate antimicrobial therapy. PMID- 10810773 TI - Molecular genetics of arrhythmias--a new paradigm. AB - The molecular genetic background of inherited cardiac arrhythmias has only recently been uncovered. This late development in comparison to other inherited cardiac disorders has partly been due to the high mortality and early disease onset of these arrhythmias resulting in mostly small nucleus families. Thus, traditional genetic linkage studies, which are based on the genetic information obtained from large multi-generation families, were made difficult. Inherited arrhythmogenic disorders can be divided into 'primary electrical disorders' (e.g., long-QT [LQT] syndrome) in which a detectable, organic heart disease is not evident, and into inherited diseases of the myocardial structure (e.g., hypertrophic cardiomyopathies) in which the arrhythmias occur combined with the structural alterations. To date, all inherited arrhythmogenic disorders in which the causative genes have been identified turned out to be channelopathies, since the genes encode channel subunits that regulate important ion currents that tune the cardiac action potential. The discovery of the genetic bases of the LQT syndrome became a new methodologic paradigm; because with the use of 'classical' genetic linkage strategies (named [positional] candidate strategies) not only the causative genes have been found, but moreover, functional components with a previously unknown but fundamental role for a normal repolarization process were discovered. Disease mutations turned out to be not only a family-specific event with a distinct phenotype and the potential of an additional diagnostic tool, but also, when expressed in heterologous expression systems, characterize the defective ion channel in a topological way and lead to a more specific understanding of ion channel function. Most, if not all, primary electrical cardiac disorders show a high genetic diversity. For the LQT syndromes, sixth disease loci and the responsible gene have been recently discovered (so-called locus or genetic heterogeneity). Within all disease genes, the mutations are spread over the entire gene (allelic heterogeneity); in addition, more than one disease mutation may be present. This complexity requires, at least, complete mutation analysis of all LQT genes before medical advice should be given. Meanwhile, genotype-phenotype correlations in large families are being used to evaluate intergene, interfamilial and intrafamilial differences in the clinical phenotype, reflecting gene specific, gene-site specific and individual consequences of a given mutation. A widespread phenotypic heterogeneity even within mutation carriers in the same family raises the importance of modifying factors and genes that are mostly unknown to date. The reduced penetrance and variable expressivity associated with the LQT mutations remain still to be explained. First insights into the complex actions of mutations are being extracted, from expression data; these preliminary results may lead to potential implications for a specific (gene-site directed) therapy. This paper discusses the current data on molecular genetics and genotype-phenotype correlations in LQT syndrome and related disorders and the potential implications for diagnosis and treatment. PMID- 10810774 TI - Molecular and electrophysiological evidence for "remodeling" of the L-type Ca2+ channel in persistent atrial fibrillation in humans. AB - Persistent atrial fibrillation (AF) is associated with shortened action potential duration (APD) and reduced atrial refractoriness. Remodeling of ion currents responsible for AP morphology has been proposed as a major mechanism in persistent AF. In the present study we investigated the activity of the cardiac L type Ca2+ channel and the mRNA transcription of the cardiac L-type Ca2+ channel subunits in patients with persistent AF compared to patients in sinus rhythm (SR). Right atrial appendages of 10 patients in SR and of 5 patients with AF were used for myocyte isolations to record L-type Ca2+ currents (ICa,L) by the patch clamp technique. Right atrial appendages of 16 patients in Sr and of 5 patients with AF served as sources for determining the mRNA expression of the L-type Ca2+ channel alpha 1c-, alpha 2/delta-, beta a-, and beta b/beta c-subunits by semiquantitative RT-PCR. ICa,L density was reduced by 70% (p < 0.001) in AF patients compared to the sinus rhythm group. Cell sizes, expressed as cell capacitance, were identical in both groups. mRNA expressions of the alpha 1c subunit and the beta b/beta c-subunits were reduced in AF patients by 18.9% (p < 0.05) and 77.7% (p < 0.005), respectively, while mRNA transcriptions of the alpha 2/delta- and the beta a-subunits were not significantly different between SR and AF patients. A decrease in the availability of functional L-type Ca2+ channels in AF patients, due to reduced alpha 1c-subunit and substantial lack of beta b/beta c-subunit transcription seems to contribute to the shortening of APD and refractory periods in AF, thereby favoring increased atrial excitation rate and perpetuation of AF. PMID- 10810775 TI - Thrombolysis and percutaneous transluminal coronary angioplasty in patients with acute myocardial infarction. AB - Acute myocardial infarction (AMI) remains a leading cause of morbidity and mortality in the developed countries. Thrombotic occlusion of a coronary artery has been shown to cause acute myocardial infarction in over 90% of the cases. Early and complete restoration of bloodflow in the infarct-related coronary artery is the principal mechanism by which reperfusion therapy improves outcomes in patients with acute myocardial infarction. Thrombolytic therapy has been shown to reduce mortality when given early after symptom onset. However, even the most effective, approved thrombolytic regimens achieve normal (so-called TIMI 3) flow in the infarct vessel at 60-90 minutes only in about half of the patients and reocclusion occurs in 5-10%. Bleeding events, especially intracranial bleedings, observed in up to 1% of the patients, are the most severe complication of thrombolysis. Primary percutaneous transluminal coronary angioplasty (PTCA) is associated with somewhat higher patency rates and significantly fewer strokes than thrombolysis, but confers a reocclusion rate of about 5-10% and it is not universally available. While smaller randomized studies suggested a significant advantage of PTCA over thrombolysis, these results could not be confirmed in the larger GUSTO IIb angioplasty study in over 1000 patients and in non-randomized comparisons in large registries. Therefore, a general mortality advantage of PTCA over thrombolysis could not be demonstrated. Primary PTCA should be preferred in patients with contraindications against thrombolysis, patients with a high risk for intracranial bleedings (age > 75 and high blood pressure on admission) and hemodynamically unstable patients. There are several approaches to improve outcome of patients with acute myocardial infarction: new fibrinolytic agents may improve early infarct related patency, single bolus administration of thrombolytics may reduce time-to-treatment, stent implantation may improve direct PTCA, enhanced thrombin and platelet inhibition may facilitate both, thrombolysis and primary PTCA, enhance reperfusion on the cellular level and reduce reocclusions and ultimately improve prognosis of patients with acute myocardial infarction. PMID- 10810776 TI - Transcoronary ablation of septal hypertrophy (TASH): a new treatment option for hypertrophic obstructive cardiomyopathy. AB - In 1991, our group started to develop a catheter interventional therapy for hypertrophic obstructive cardiomyopathy (HOCM). The new concept was proposed in 1994. It is based on the conventional PTCA technique with the aim of inducing an artificial myocardial infarction by instillation of 96% ethanol into the most proximally situated septal branch of the left anterior descending coronary artery. This leads to a subaortic contraction disorder with subsequent decrease of the intraventricular pressure gradient, shrinkage of the hypertrophied septal bulge and widening of the outflow tract ("therapeutic remodeling"). The subaortic defect is small and well demarcated as assessed by left ventricular angiography, transesophageal echocardiography and 18 F-glucose positron emission tomography. The term transcoronary ablation of septum hypertrophy (TASH) was suggested. Our patient cohort that now comprises 215 therapeutic procedures in 187 patients underwent a large variety of prospective studies (maximum follow-up 4.5 years) including invasive controls at regular intervals, investigation of hemodynamics at rest and at exercise, transesophageal and transthoracic echocardiography, Doppler echocardiography during bicycle exercise, electrophysiologic testing, Holter monitoring and measurement of myocardial metabolism and perfusion, assessment of microembolic events by transcranial Doppler sonography and histological examinations. This article gives an overview and reports our increasing experience in applying TASH. The following post-TASH findings were obtained: significant hemodynamic and clinical improvement at rest and at exercise, decrease of septum thickness, increase of outflow tract area and decrease of induced ventricular tachycardia. There were well-demarcated, histologically atypical subaortic myocardial defects, no microembolic events, abnormal early peak of infarct related enzymes, and no change of baroreflex sensitivity. Pre-/post-TASH evaluations of the patients should be based in particular on clinical symptoms correlated to the intraventricular gradient measured by bicycle exercise Doppler echocardiography and to outflow tract area as assessed by transesophageal echocardiography. Since 1994, as a roughly estimate, worldwide 1000 patients in 20 countries have been treated. According to published articles, abstract presentations and workshops, TASH consistently leads to a pronounced clinical and hemodynamic benefit for patients with HOCM. TASH has become an established technique. At least in centers with a high level of expertise, it is no longer experimental but a routinely performed alternative to surgical treatment for HOCM, i.e., the previous gold standard of therapy. Of course, patient outcome needs further careful clinical and prognostic evaluation. With respect to complications, TASH appears to be superior to surgery (transaortic septal myectomy) for HOCM. Like surgical treatment, TASH is currently indicated in critically ill patients with typical HOCM (subaortic form), who exhibit with drug refractory symptoms, including patients, who preferred DDD pacemaker therapy as a first therapeutic step but in whom this produced no subsequent clinical benefit. PMID- 10810777 TI - Positron emission tomography and the changing paradigm in coronary artery disease. AB - While lowering cholesterol with HMG-CoA reductase inhibitors appears to affect little if at all the severity of coronary artery stenoses, at least over the time periods studied, it markedly reduces cardiac morbidity and mortality. The beneficial effects of these agents have therefore been attributed to plaque stabilization and improved coronary vasomotor function. This then has shifted the emphasis of detection and treatment of coronary artery disease from the assessment of structural to functional alterations. With PET based measurements of regional myocardial blood flow, the function of the coronary circulation in humans can now be evaluated noninvasively. Multiple studies with this approach reported diminished hyperemic flow responses in patients without hemodynamically significant coronary stenoses but with coronary risk factors. Furthermore, lipid lowering with HMG-CoA reductase inhibitors was consistently found to enhance hyperemic flow responses to pharmacologic vasodilation as an index of the integrated function of the human coronary circulation. Recent studies with PET based measurements of blood flow target specifically the endothelial dependent coronary vasomotion and revealed abnormalities in chronic smokers and in postmenopausal women, that is, after estrogen withdrawal. Conversely, L-arginine supplementation in smokers and hormone replacement therapy in postmenopausal women without coronary risk factors normalized endothelial dependent flow responses. These observations suggest that endothelial dysfunction as a pivotal event early in the development of coronary atherosclerosis can be identified noninvasively with PET which will also be useful for measuring responses to lifestyle modification and pharmacologic treatment for improving coronary circulatory function. PMID- 10810778 TI - Medical progress and rationing of cardiological services. AB - Due to the financial restrictions imposed on all national health services, the problem of allocating medical resources has gained new economical and political dimensions. Although each patient expects optimal treatment, the rationing of medical services make it increasingly difficult to provide the most adequate therapy. Health politicians and medical doctors have all advocated that health expenditures can be reduced by effective prevention of coronary heart disease, one of the main cost factors for all health services. Due to the introduction of statins, primary and secondary prevention of coronary heart disease can now be accomplished effectively. However, such treatment does not induce cost reduction, but rather an increase in costs. This applies predominantly to primary prevention. Treatment of all who would benefit from lipid lowering by statins would be prohibitively expensive for all national health care systems. PMID- 10810779 TI - [Genetic aspects of the etiology of arrhythmia]. AB - Cardiac arrhythmias are common causes of morbidity and mortality in clinical medicine. Much has been learned about cellular mechanisms of arrhythmogenesis in the past but genetic components have only recently been recognized for some heritable forms of arrhythmias. The long QT syndrome and the Brugada syndrome are both caused by molecular defects in ion channel proteins. Cardiac arrhythmias can also be associated with structural heart diseases. For example, sinus node dysfunction or AV-block can precede some forms of inherited dilated cardiomyopathy. A distinct genetic form of hypertrophic cardiomyopathy is associated with the Wolff-Parkinson-White syndrome and maps to chromosome 7q35. Arrhythmogenic right ventricular cardiomyopathy has a strong genetic basis and often manifests with ventricular tachycardia. Atrial fibrillation can also occur as familial disease and may be allelic with dilated cardiomyopathy as both diseases can be closely linked to chromosome 10q2. PMID- 10810781 TI - [Acute myocardial ischemia and ventricular arrhythmias in the pathogenesis of sudden cardiac death in coronary disease]. AB - There is increasing evidence for a fatal interaction of myocardial ischemia, ventricular arrhythmias and sudden cardiac death in some patients with coronary artery disease. Evidence comes from autopsy studies, from the evaluation of patients who survived an episode of sudden cardiac death, from follow-up data of these patients either treated or not by revascularization therapy and/or an implantable cardioverter-defibrillator and indicate that reducing the individual ischemic burden will be beneficial to reduce the incidence of sudden cardiac death. Studies in patients with stable and especially with unstable angina using Holter monitoring could demonstrate that there is a close and causal relationship between myocardial ischemia inducing or aggravating life-threatening ventricular arrhythmias and sudden cardiac death particularly in patients with unstable and postinfarction status. This review summarizes some of our clinical knowledge on this topic and indicates that preventive strategies for myocardial ischemia are the antiarrhythmic treatment of choice in patients with severe coronary artery disease and patients with evidence or at risk for ischemic proarrhythmia. PMID- 10810780 TI - [Ion channels and arrhythmias]. AB - Changes in ionic currents through ion channels of the myocardial cell membrane have to be regarded as main cause of cardiac arrhythmias. Three basic arrhythmogenic mechanisms are responsible for the initiation of tachyarrhythmias: 1. The disturbance of normal automaticity in cardiac pacemaker cells dependent on the currents If, ICa-L, ICa-T or IK-ACh,Ado and the occurrence of abnormal automaticity in atrial and ventricular working myocardium based on the currents ICa-L, INa, IK, IK1 or IK-ACh,Ado. 2. Triggered activity which may be recognized by the appearance of early (EAD) or late afterdepolarizations (LAD). EAD are mainly due to inhibition of the outward currents IKr and IKs and are favoured by an increase in the inward currents INa and ICa-L, respectively. Typical arrhythmias are torsade de pointes occurring during treatment with K(+)-channel inhibitors (e.g. sotalol) or in patients with QT-syndrome. LAD may be observed during Ca(2+)-overload of the myocardial cell (digitalis intoxication, catecholamines) and are based on the transient inward current Iti, which is build up by the participation of the currents INa/Ca, INS and ICa-L. 3. Reentry mechanisms are the most frequent cause of tachyarrhythmias. They originate in an anatomically defined excitation circle with unidirectional block. Na(+)- and Ca(2+)-channel dependent disturbances of conduction with long excitable gap may be distinguished from Na(+)-channel dependent disturbances of conduction and refractory period with short excitable gap. Interruption of reentry is possible in the first case by depression of conduction and excitability (Na(+)- or Ca(2+) channel blockers), in the second case by increase in refractory period (K(+)- or Na(+)-channel blockers). PMID- 10810782 TI - [Inflammation of the myocardium as an arrhythmia trigger]. AB - In patients with acute or chronic myocarditis, arrhythmias are a common and often the only clinical symptom in the natural course of the disease. The potentially malignant tachy- and bradyarrhythmias are of particular significance in the differential diagnosis of sudden cardiac death in myocarditis. Factors responsible for the increased incidence of cardiac arrhythmias are structural changes, parameters of ventricular dynamics and vascular changes. On the one hand, inflammatory processes in the cardiac myocytes and interstitium can lead directly to fluctuations in membrane potential. Fibrosis and scarring of the myocardial tissue and secondary hypertrophy and atrophy of the myocytes favor the development of ectopic pacemakers, late potentials and reentry as a result of inhomogeneous stimulus conduction. Furthermore, parameters of ventricular dynamics such as increased wall tension, increased myocardial oxygen consumption and diminished coronary reserve in the case of disturbed systolic or diastolic left ventricular function also contribute to the increased incidence of arrhythmias. Lastly, vascular factors can further increase the arrhythmogenicity of the inflamed myocardium through the disturbance of micro- and macrovascular perfusion and the resulting myocardial ischemia. Non-invasive rhythmological evaluation by 24 h Holter ECG, measurement of ventricular late potentials and heart rate variability can be used for orienting risk stratification of the at risk patient with myocarditis. Programmed atrial and ventricular electrophysiological stimulation also has a relatively high predictive value for spontaneous ventricular tachyarrhythmias. It should be emphasized that, at the present time, optimal electrophysiological parameters with a high predictive value do not exist. In a selected patient population, immunosuppressive therapy in addition to conventional antiarrhythmic therapy can lead to the reduction or complete suppression of spontaneous and inducible arrhythmias. Nevertheless, in the interim, further precautionary antiarrhythmic measures such as serial antiarrhythmic treatment, VT ablation and ACID implantation are necessary in patients with malignant cardiac arrhythmias. Right ventricular myocardial biopsy for demonstration or exclusion of myocarditis is an important additional examination which can improve the differential diagnosis and treatment of patients with cardiac arrhythmias of unclear etiology. PMID- 10810783 TI - [Arrhythmia risk in left ventricular hypertrophy]. AB - The incidence of supraventricular and ventricular arrhythmias in patients with arterial hypertension is up to 96% and is about 10 times higher than in normotensives. Predictors for an increased ventricular arrhythmogenic risk are left ventricular hypertrophy (LVH), impaired left ventricular function with enlarged end diastolic and end diastolic volumes as well as late potentials which in case of LVH increase from a 7% to 18% incidence. Especially the Simson criteria fQRS and RMS seem to characterize patients at risk. In addition a longer duration of hypertension in conjunction with a higher muscle mass index and a larger amount of couplets and non-sustained ventricular tachycardias, documented by Holter recording, are determinants of life threatening arrhythmias. In addition, an increased ventricular vulnerability in electrophysiological study significantly depends on left ventricular hypertrophy. Regression of LVH goes along with a decreased rate of ventricular extrasystoles. We therefore hypothesize that by pharmacological regression of hypertrophy the prevalence of complex arrhythmias decreases. PMID- 10810784 TI - Baroreflex sensitivity as a new marker for risk stratification. AB - As the arterial baroreflex importantly contributes to modulation of the autonomic influences on the heart and thereby arrhythmogenesis, baroreflex sensitivity has been used as a measure of the interaction between sympathetic and parasympathetic activities at the cardiac level. The most widely applied technique both in the experimental and clinical setting is the measurement of the heart rate slowing in response to a blood pressure rise induced by small intravenous boluses of phenylephrine. Baroreflex sensitivity is expressed as ms/mmHg and prevailing vagal reflexes are reflected by the wider R-R interval lengthening. The experimental evidence that the occurrence of ventricular fibrillation was inversely related to baroreflex sensitivity, opened the way to clinical studies. The ATRAMI (Autonomic Tone and Reflexes After Myocardial Infarction) trial has definitely demonstrated not only that a depressed baroreflex sensitivity (< 3 ms/mmHg) is a strong risk factor for cardiac death, but also that the information gained by the analysis of autonomic markers adds to the information obtained by better recognized measures of cardiovascular outcome such as left ventricular function and ventricular arrhythmias. The value of a depressed baroreflex sensitivity as a risk stratifier is meaningful in patients below age 65 in combination of a simultaneously depressed left ventricular ejection fraction. In these patients, the analysis of autonomic activity might be of value in the identification of patients who may need an implantable automatic defibrillator for primary prevention of sudden cardiac death. PMID- 10810785 TI - [Heart rate variability and chemoreflex sensitivity. Proved and new methods in risk prediction of malignant arrhythmias]. AB - For the analysis of a disturbed autonomic function as a risk predictor for ventricular tachyarrhythmias, tonic and phasic procedures are available. The heart rate variability as a tonic procedure shows significant differences between patients with an increased risk of malignant arrhythmias and patients without increased risk. This can be demonstrated in patients with survived myocardial infarction, dilated cardiomyopathy and congestive heart failure. But the positive predictive value amounts only to about 50%. The chemoreflex sensitivity as a new phasic method represents a new possibility for the evaluation of a dysfunction of autonomic reflex arches. It is reduced due to a decreased left ventricular function and increasing age. Furthermore, it shows significant differences between patients with ventricular arrhythmias and patients without. The predictive accuracy concerning malignant ventricular arrhythmias in a population of 60 patients in the chronic postinfarction stadium amounts to 55%, the relative risk to 7.6. Thus, this method shows a high predictive power, but more investigations in larger patient cohorts are necessary to corroborate these results. PMID- 10810787 TI - [Current classification of anti-arrhythmia agents]. AB - Antiarrhythmic drugs can be divided into four Vaughan Williams classes (I-IV) according to defined electrophysiological effects on the myocardium. Thus, the Vaughan Williams classification also coincides with the main myocardial targets of the antiarrhythmics, i.e., myocardial sodium-, potassium-, and calcium channels or beta-adrenergic receptors. A more detailed characterization which is also based on the myocardial targets of a drug is given by the "Sicilian Gambit" approach of classification. Nevertheless, the appropriate drug for the management of a given clinical arrhythmia has to be chosen according to the electrophysiological effects of the respective drug. A main determinant of the antiarrhythmic or proarrhythmic properties of a drug is the frequency dependence of its electrophysiological effects. The sodium-channel blockade induced by class I substances is enhanced with increasing heart rates. Thus, class-I antiarrhythmics can be subclassified as substances showing a more exponential, an approximately linear, or rather saturated block-frequency relation. Class-III antiarrhythmics (potassium-channel blockade) can be further differentiated according to the component of the delayed rectifier potassium current (IK) which is inhibited by a drug. Class-III drugs inhibiting selectively the rapidly activating and deactivating IKr component exhibit a marked reverse rate dependence, i.e., the drug induced prolongation of the cardiac action potential is minimized at high rates. On the other hand, during bradycardia the pronounced action potential prolongation may cause early afterdepolarizations and triggered activity leading to torsades de pointes arrhythmias (acquired QT syndrome). Class III substances inhibiting the slowly activating IKs component are currently under investigation and are expected to show a direct rate dependence. Experimental data available so far point to an action potential prolonging effect at least independent of rate. However, it is uncertain whether proarrhythmic effects can be thus avoided, especially in light of the fact that one form of congenital QT syndrome (LQT1) seems to be linked to dysfunction of the IKs-channel. PMID- 10810786 TI - [Microvolt level T wave alternans: a new marker for noninvasive risk stratification]. AB - Prospective identification of patients with structural heart disease who could profit from prophylactic ICD therapy is hampered by the low predictive power of the currently available risk stratification parameters. Microvolt T wave alternans measured noninvasively is a new promising parameter to assess impaired ventricular repolarization which has been associated with an increased incidence of ventricular tachyarrhythmias. T wave alternans is rate-dependent; to induce alternans, heart rate may be increased by atrial stimulation during invasive EP testing or noninvasively by exercise stress testing. The first clinical validation with respect to prediction of inducibility of ventricular tachyarrhythmias and of arrhythmic events during follow-up in patients undergoing invasive EP testing was reported in 1994. Subsequently, a good concordance between the results of invasive and noninvasive assessment of T wave alternans was demonstrated by our group. The first prospective evaluation of the noninvasive alternans measurement using submaximal exercise testing was performed in patients surviving prehospital ventricular fibrillation or sustained ventricular tachycardia referred to our institution. The occurrence of T wave alternans in this patient population was predictive of future tachyarrhythmic events with subsequent appropriate ICD therapy. The results of the currently performed prospective trials in various patient populations will help to establish the utility of T wave alternans assessment as a risk stratifier in clinical practice. PMID- 10810788 TI - [Drug and electrical therapy of supraventricular tachyarrhythmias]. AB - This article reviews current pharmacological and electrical approaches to the restoration of sinus rhythm in patients who suffer from atrial fibrillation and atrial flutter. Spontaneous conversion to sinus rhythm occurs in a high proportion of atrial fibrillation of < 24 h duration. Among patients presenting with atrial fibrillation, which was clinically estimated to have lasted < 48 h, the likelihood of cardioversion-related clinical thromboembolism is low, which supports the current practice of early cardioversion without anticoagulation. Antiarrhythmic drugs effective in terminating atrial fibrillation of short duration are those which possess class IA, IC and III properties. Electrical transthoracic cardioversion by using different electrode positions and additional pressure over the electrodes during shock delivery is a highly effective and well tolerated method in restoration of sinus rhythm even in patients under conscious sedation. Immediate spontaneous reinitiation of atrial fibrillation can occur in a significant proportion of patients undergoing electrical cardioversion and can be reduced after a pretreatment with antiarrhythmic drugs. In patients with failed external cardioversion, internal low energy cardioversion offers an effective option for restoring sinus rhythm. After cardioversion in a high proportion of patients antiarrhythmic drugs are necessary to prevent atrial fibrillation from recurring. A serial cardioversion approach can prevent the evolution of permanent atrial fibrillation in a subgroup of patients. Overdrive atrial pacing is an effective and minimally invasive procedure for termination of atrial flutter. The acute administration of class IA, IC and III antiarrhythmic drugs increases the success rate of this method in restoring sinus rhythm. PMID- 10810789 TI - [Risk stratification after myocardial infarct]. AB - In industrialized countries the rate of sudden cardiac death remains unchanged. The most frequently encountered structural heart disease in these patients is coronary artery disease. Despite the era of thrombolytic therapy of acute myocardial infarction patients carry an increased risk of sudden cardiac arrhythmogenic death within a time period of one to two years following the acute event. Therefore, risk stratification post-MI before patient discharge is furthermore mandatory. The spectrum of non-invasive techniques for risk stratification includes the clinical risk profile, measurement of left ventricular global function (LV ejection fraction), the resting ECG (QT dispersion), an ECG stress test (detection and severity of myocardial ischemia), ambulatory ECG monitoring (number and type of ventricular arrhythmias), surface high resolution ECG (detection of ventricular late potentials), measurement of T wave alternans (TWA, alternans ratio), and measurements of the activity and balance of the autonomous nervous system (heart rate variability, baroreflex sensitivity = BRS). Programmed ventricular stimulation (PVS) serves as an invasive risk stratification technique (detection of an arrhythmogenic substrate). The prognostic power of the non-invasive techniques is limited; in general, the prognostic value of a negative test is reasonably high (90 to 100% depending on the test used), whereas the prognostic value of a positive test is rather low (4 to 42% depending on the test used). Combining several non-invasive tests may significantly improve the positive predictive value above 50%, but this goes along with a significant decreases of sensitivity below 50%. Therefore, a combination of several non-invasive tests (detection and exclusion of a large number of low-risk individuals) with the invasive method of PVS (detection of an arrhythmogenic substrate, i.e. a high-risk patient) seems reasonable, as has been convincingly shown by several smaller prognostic studies. PMID- 10810790 TI - [Catheter ablation of supraventricular tachyarrhythmias: experimental findings]. AB - In the majority of cases, atrial fibrillation seems to depend on multiple, simultaneously active reentrant circuits. Their pathlength is defined by local refractory periods and conduction velocities. Active circuits experience constant changes in number, location and dimension. Ablative interventions targeting individual circuits are, therefore, not likely to be successful. Current approaches aim at multiple, electrically isolated atrial segments providing insufficient space for the given minimal size of potential reentrant circuits. Pharmacologic interventions either result in conduction delay and subsequent conduction block, or in prolongation of local refractoriness increasing the size of existing circuits. Creation of isolated atrial segments by endocardial catheter ablation implies the feasibility of extended, contiguous transmural lesions. Mapping studies in healthy dogs demonstrated that contiguous anatomic lesions are very hard to achieve with conventional catheter techniques. Functional conduction blocks, however, may be obtained. Furthermore, application of radiofrequency energy induces prolongation of local refractoriness even at a distance from the actual anatomic lesion. Thus, in the interventional therapy of atrial fibrillation, functional effects of radiofrequency ablation might at least serve to facilitate the strenuous implementation of the isolation concept. PMID- 10810791 TI - [Catheter ablation of accessory pathways and atrioventricular nodal reentry tachycardia]. AB - Radiofrequency current catheter ablation has gained acceptance as primary long term therapy for patients with symptomatic accessory pathways and symptomatic atrioventricular nodal reentrant tachycardia (AVNRT) with frequent recurrences. In both arrhythmias, curative treatment is possible in more than 90% of cases at a low complication rate although an incidence of about 1% complete AV block after slow pathway ablation has to be taken into account when this therapy is considered. The recurrence rate is 3-10% for accessory pathways and 0-15% for AVNRT. The high success rate of catheter ablation has already led to a shift in the indications for the procedure where the percentage of patients with accessory pathways is decreasing and there is an increase of patients with AVNRT and newer indications (atrial flutter, focal atrial tachycardias). PMID- 10810792 TI - Radiofrequency ablation of atrioventricular nodal reentrant tachycardia: mechanisms and recurrence rate. AB - Radiofrequency catheter ablation is the treatment of choice in atrioventricular nodal reentrant tachycardia. Electrophysiologic investigations in 623 patients revealed eight mechanisms of tachycardia ablation: Ablation of fast (I) or slow (II), modification of fast (III) or slow AV nodal pathways (IV), modification of both pathways (V), ablation of fast and modification of slow (VI), ablation of slow and modification of fast pathways (VII) and ablation of both pathways (VIII). The criteria of diagnosis of these eight mechanisms of tachycardia ablation are described. Follow-up showed fewer relapses in patients with ablation (0-2%) in comparison to patients with modification of a single AV nodal pathway (8-12%). Alteration of both pathways includes an increasing risk of total AV nodal block, which occurred in 7 patients (1.1%). Detailed analysis of the mechanism of catheter ablation is recommended in all patients after radiofrequency current delivery for AV nodal reentrant tachycardia to estimate the risk of relapse during follow-up or development of total AV block in the particular patient in case of a further ablation procedure. PMID- 10810793 TI - [Non-drug treatment methods in ventricular rate control in atrial fibrillation: high-frequency catheter ablation and catheter modification of the AV node]. AB - AV nodal ablation with pacemaker implantation nor AV nodal modification is the therapy of the choice in patients with permanent or paroxysmal atrial tachyarrhythmias. Both methods of treatment are limited to patients with drug refractory atrial tachyarrhythmias. Therapy indication requires the complete knowledge of the current clinical status and medical history of the patient regarding the time course of the disease and the results of previous treatments. Both methods are effective and palliative treatments which may not modify the underlying arrhythmias. Neither treatment showed an improvement in patients prognosis; therefore a restrictive indication is mandatory. PMID- 10810794 TI - [How to ablate: atrial tachycardia]. AB - The ectopic atrial tachycardia is diagnosed in approx. 10 to 15% of supraventricular tachycardias. Since medical therapy is often shown to be ineffective, ablative procedures became especially important in this disease. Localization of the arrhythmia foci seems to be concentrated at anatomical preformations like the crista terminalis, right or left atrial appendage orifice or at the superior pulmonary veins. The P-wave vector in the surface ECG is predictive for focus localization: a positive or biphasic P-wave in a VL for right atrial foci, a positive P-wave in V1 for left atrial foci. Catheter ablation therapy is also shown to be very effective in this kind of supraventricular tachycardia. PMID- 10810795 TI - [High frequency catheter ablation of accessory atrioventricular pathways]. AB - Radiofrequency catheter ablation has established as the first line therapy for the curative treatment of patients with accessory pathways. For left-sided accessory pathways, the retrograde approach over the aortic valve is commonly used for ablation of the ventricular insertion. For right-sided and septal accessory pathways, the atrial insertion is usually approached from the right atrium. Atrioventricular accessory pathways irrespective of the exact localization can be successfully ablated in more than 90-95% of all cases. Severe complications associated with the ablation procedure are rare and occur in approximately 2-3% of patients treated. The recurrence rate after successful ablation is approximately 5-10%. Recurrences of accessory pathway conduction occur almost exclusively within the first 3 months following successful ablation whereas late recurrences are rare. Because of the favorable efficacy--risk profile, radiofrequency catheter ablation can be recommended as the first line therapy to all symptomatic patients with accessory atrioventricular pathways. PMID- 10810796 TI - Mahaim fibers: electrophysiologic characteristics and radiofrequency ablation. AB - Mahaim fibers that exhibit decremental atrioventricular (AV) node-like conduction properties are unusual, comprising < 3% of accessory pathways. They are involved in antidromic AV reciprocating (Mahaim) tachycardia with a left bundle branch block morphology. There is a frequent association of Mahaim pathways with Ebstein's anomaly, additional AV accessory pathways (Kent) and dual AV nodal pathway conduction. The majority of these pathways are long right atriofascicular pathways capable of only anterograde conduction. Radiofrequency (RF) ablation of right atriofascicular pathways guided by a distinct Mahaim potential recorded at the anterolateral-to-posterolateral tricuspid annulus or in the right ventricular free wall is safe and highly effective. True nodoventricular or nodofascicular Mahaim pathways usually also capable of only anterograde conduction appear to be rare and are associated with pre-excited tachycardia of left bundle branch block morphology not distinguishable from the preexcitation pattern of right atriofascicular pathways. In these pathways, a reliable method for mapping and safe RF ablation is lacking because no Mahaim potential could be recorded from the tricuspid annulus in the right midseptal region where ablation may result in elimination of both the slow AV nodal pathway and Mahaim conduction. Fasciculoventricular Mahaim pathways have never been reported to be involved in a tachycardia circuit. At our institution, successful RF ablation of unusual Mahaim pathways (concealed nodoventricular and nodofascicular and left anterograde atriofascicular) was performed without impairing the normal AV conduction system. PMID- 10810797 TI - Radiofrequency ablation of atrial tachycardia and atrial flutter. AB - Atrial endocardial mapping defines the activation pattern during regular atrial arrhythmias. The response to pacing (entrainment mapping) yields additional information about the mechanism and location of the circuit. Regarding radiofrequency ablation, the regular atrial tachycardias may be classified in two broad patterns: 1) macroreentrant tachycardias, characterized by circular activation, including typical atrial flutter and reentry around scars, and 2) focal tachycardias, characterized by radial activation from a small myocardial area. Catheter ablation of right atrial macroreentrant circuits and focal tachycardias are now standard procedures. The target for ablation of focal tachycardia is the point of earliest activation, and single application is effective in most cases. Ablation of macroreentrant tachycardias requires identifying a narrow isthmus inside the circuit, and linear ablation is needed in most cases. The goal of radiofrequency ablation for typical atrial flutter is to interrupt conduction across the inferior vena cavatricuspid valve isthmus. The procedure is highly effective and safe, although recurrences of the arrhythmia are not rare. Incisional macroreentrant tachycardias after surgical correction of congenital heart diseases and primary cardiac tumors may also be ablated identifying critical isthmuses in the circuit by a combination of mapping and entrainment techniques. Left atrial macroreentry is much less known and at present more information is needed before catheter ablation becomes a standard procedure. PMID- 10810798 TI - [Theory and practical approaches in catheter ablation of atrial flutter]. AB - Atrial flutter can be understood as atrial tachycardia due to a single intraatrial macroreentrant circuit that is determined by fixed or functional boundaries. In various types of atrial flutter, radiofrequency ablation became an established curative therapy. During the course of an ablation procedure, initially, the reentrant circuit has to be determined, e.g. by activation and entrainment mapping. Subsequently, the boundaries have to be identified. By connecting two appropriate boundaries with a linear lesion, the intraatrial reentrant circuit can be inhibited. Finally, it should be proven that the linear lesion results in a complete line of conduction block. Doing so, the acute and long-term results of atrial flutter ablation are comparable to those of other supraventricular tachycardias. PMID- 10810799 TI - [Endocardial mapping and high frequency catheter ablation of ventricular tachycardia after myocardial infarction]. AB - Recurrent ventricular tachycardia in the setting of remote myocardial infarction are frequently resistant to antiarrhythmic drug treatment. Endocardial mapping and ablation is feasible in case of hemodynamically tolerable and reproducibly inducible forms. Identification of critical components of the reentrant circuit is mainly guided by entrainment mapping and the analysis of the post-pacing interval. The emergence of multiple types of ventricular tachycardia is a common limitation of the procedure. Ventricular tachycardia can be acutely abolished by radiofrequency current ablation in 60-70% of cases when only single forms are present. This success rate is substantially lower in case of multiple tachycardia morphologies. The incidence of tachycardia recurrences varies from 20-30%. The overall mortality during follow-up is increased due to progressive heart failure and the occurrence of rapid ventricular tachyarrhythmias. Catheter ablation has been shown to be a useful tool for the treatment of clusters of ventricular tachycardia following implantation of a cardioverter-defibrillator. Furthermore, this method can be life-saving in the setting of incessant forms. Currently, catheter ablation represents an adjunctive treatment to antiarrhythmic drugs and the implantation of a cardioverter-defibrillator. Improvement of mapping and ablation technologies may help to further increase the efficacy of this treatment strategy in the near future. PMID- 10810800 TI - [New ablation techniques in ventricular tachycardia]. AB - Catheter ablation of ventricular tachycardia (VT) has proven to be effective in only a minority of VT patients. Therefore there is great interest in the development of new mapping and ablation techniques. New mapping procedures include multielectrode catheter mapping via the epicardial cardiac arteries and veins and the multielectrode basket catheter. Computerized non-contact mapping offers a three-dimensional approach in defining the activation sequence during VT. New ablative techniques are "cooled" radiofrequency energy application and microwave ablation which allow deeper and larger lesions. Already used in clinical practice are the application of laser energy and chemical ablation. New developments are ultrasound-, cryo- and thermal ablation. It can be suspected that some of these new achievements will lead to better results in VT ablation. PMID- 10810801 TI - [Non-contact mapping: a simultaneous spatial detection in the diagnosis of arrhythmias]. AB - The non-contact mapping device (Ensite, Endocardial Solution) enables a simultaneous, spatial detection of the myocardial depolarization. An isopotential map is visualized in the 3D reconstruction of the cardiac chamber. The analysis of focal premature beats, of unstable arrhythmias and of macro reentry mechanisms is possible. The current article demonstrates the use of the non-contact mapping device for analysis and ablation of atrial flutter, premature beat associated atrial fibrillation and of ventricular tachycardias. PMID- 10810802 TI - [Complications and risk of high frequency catheter ablation of tachycardiac arrhythmias]. AB - Radiofrequency catheter ablation has established itself as a first line therapy for the curative treatment of many patients with supraventricular or atrioventricular tachycardias and also for selected types of ventricular tachycardia. The success rates of catheter ablation of various types of cardiac arrhythmias are impressively high. Procedure related complications can be attributed to the invasive nature of the technique (e.g., bleeding or other vascular complications, radiation exposure) but may also occur as a specific complication related to the type of intervention performed (e.g., complete AV block following attempted modification of the AV-node). In patients undergoing radiofrequency ablation procedures, radiation exposure carries a small but measurable risk of malignancy and hereditary disorders. The risk of fatal malignancy has been calculated to be approximately 1/1000 per hour of fluoroscopy and the risk of significant hereditary disorders approximately 10 per 1 million live births per hour fluoroscopy time. However, it is important to realize that these risks are age and sex dependent being higher in young and/or female patients. For the physician performing catheter ablation procedures no significant risks related to fluoroscopy exposure may be expected as long as all established tools for protection are used. Based on the results of large single center studies and multicenter investigations, complications during or after radiofrequency catheter ablation of supraventricular or atrioventricular arrhythmias may occur in 4-5% of cases. Severe complications (life threatening or permanently disabling complications) may occur in approximately 1-2% of patients treated. In patients undergoing ablation of ventricular tachycardia, a higher incidence of total procedure related complications between 5-7% and severe complications (3-4%) may be expected. The higher incidence of complications in patients with ventricular tachycardia when compared to catheter ablation of supraventricular or atrioventricular tachycardia may be explained by the fact that many patients with ventricular tachycardia suffer from severe cardiovascular disease. PMID- 10810803 TI - [14-year experience with implantable cardioverter/defibrillators: determination of prognosis and discharge behavior]. AB - BACKGROUND: The treatment of life threatening ventricular arrhythmias with implantable cardioverter/defibrillators (ICD) has become the therapy of choice; the survival benefit of ICD treatment compared to drug therapy in patients with aborted sudden cardiac death (SCD) and hemodynamically unstable ventricular tachycardia has been proven. In addition for the primary prevention of SCD in high risk patients, ICD therapy is gaining growing acceptance. PATIENTS AND METHODS: We analyzed the long-term follow-up of 274 consecutive patients (211 male, 63 female, age 59 +/- 12 years, left ventricular ejection fraction 39 +/- 15%) provided with an ICD between 1984 and 1998. The aim of the study was to ascertain the survival rate in different subgroups and to discover determining factors of ICD discharge and prognosis. RESULTS: Long-term survival probability at 10 resp. 14 years was 84 resp. 65% for the total collective, and the freedom of event probability (neither shocks nor antitachycardiac pacing from the ICD) to 28% each. The risk to die from SCD was below 3% over time. The most pronounced differences regarding prognosis ensued from dividing the collective into heart insufficiency stages. Thus in NYHA class I and II versus III and IV, the cumulative event rate was 61% vs 82% at 5 years, and survival rate amounted to 94 vs 63% at 5 years and 87% vs 30% at 14 years (p < 0.001). Calculating the relative benefit of ICD therapy survival benefit provided by the ICD was shown to decrease significantly after 5 years for patients in NYHA class III/IV, while it increased progressively for patients in NYHA class I/II up to 10 years. Additional determinants of prognosis and ICD discharge rate were identified left ventricular ejection fraction, age and tendency for the basic cardiac disease, however neither the result of electrophysiological testing nor the results of non invasive risk stratification. In patients with ischemic heart disease, revascularization procedures improved prognosis only in tendency, while the effect of ICD therapy was significant. In patients with the non-obstructive form of hypertrophic cardiomyopathy ICD, discharges occurred in about 50% of patients; in contrast patients with surgical myectomy for obstructive cardiomyopathy showed no events during follow-up. In patients with chronic inflammatory heart disease and normal left ventricular function (LVF), a very low event rate was expected if patients were treated by immunosuppressive drugs. Patients with dilated cardiomyopathy did not differ from patients with ischemic heart disease with respect to prognosis and ICD discharge rate. CONCLUSION: Significant determinants of prognosis and ICD discharge rate are left ventricular function, age and with limitations the basic cardiac disease. In contrast to patients with better LVF relative survival benefit decreases significantly after 5 years in patients with a worse LVF. Patients with aborted SCD and preserved LVF experience half the ICD discharges compared to patients with poor LVF and gain at the same time a normalization of life expectancy. Causative treatment of the basic disease has an impact on the overall prognosis and event rate, but should in general not influence the decision for IDC implantation in high risk patients. PMID- 10810804 TI - [Atrial defibrillator]. AB - Atrial fibrillation (AF) is a frequent and costly health care problem representing the most common arrhythmia resulting in hospital admission. Total mortality and cardiovascular mortality are significantly increased in patients with AF compared to controls. In addition to symptoms of palpitations, patients with AF have an increased risk of stroke and may also develop decreased exercise tolerance and left ventricular dysfunction. All of these problems may be reversed with restoration and maintenance of sinus rhythm. External electrical cardioversion has been a remarkably effective and safe method for termination of this arrhythmia. Originally described by Lown et al. in 1963, it has been a well accepted mode of acute therapy. However, this technique requires general anesthesia or heavy sedation. Internal atrial defibrillation has been evaluated as an alternative approach to the external technique for over 2 decades. Recent studies have shown that low-energy internal atrial defibrillation using biphasic shocks is an effective and safe means in restoring sinus rhythm in patients with AF and should be considered especially in patients in whom external cardioversion attempts have failed. Implantable Atrial Defibrillator: Recently, a stand alone IAD, the Metrix System (models 3000 and 3020), has entered clinical investigation. Atrial defibrillation is accomplished by a shock delivered between electrodes in the right atrium and the coronary sinus. The right atrium lead has an active fixation in the right atrium. The coronary sinus lead has a natural spiral configuration for retention in the coronary sinus, and can be straightened with a stylet. Both leads are 7 French in diameter and the defibrillation coils are each 6 cm in length. The electrodes may be placed using separate leads, or very soon by using a single bipolar lead. A separate bipolar right ventricular lead is used for R wave synchronization and post shock pacing. The Metrix defibrillator can be used to induce AF by using R wave synchronous shocks and can store intracardiac electrograms (EGMs) for up to 2 minutes from the most recent 6 AF episodes. The device can be programmed into one of the following operating modes: fully automatic, patient activated, monitor mode, bradycardia pacing only, and off. As AF is not life-threatening, in the automatic mode the device is only intermittently active in detecting and treating AF, and this "sleep wake-up" cycle interval is programmable. The device employs extensive processing both for detection and R wave synchronization. In April 1996, the phase I Metrix multicenter clinical trial was started. As of May 1997, a total of 51 Metrix systems had been implanted as part of the phase I multicenter clinical trial. Preliminary data suggest that both defibrillation thresholds and electrograms are stable over time (implant to 3 months). Detection accuracy has been excellent (100% specificity, 92.3% sensitivity) and there have been no errors of R wave selection for synchronization. No proarrhythmias have resulted from over 3700 shocks delivered. The device is effective in electrically converting 96% of the spontaneous episodes of AF. In 27% of episodes several shocks were required because of early recurrence of AF. In 5 patients, the atrial defibrillator was removed: 2 infections, 1 cardiac tamponade, 1 permanent loss of telemetry, 1 patient required His-Bundle ablation because of frequent episodes of drug refractory AF with rapid ventricular response. Initial clinical experience under controlled conditions with the Metrix system suggests that the implantable atrial defibrillator may offer a therapeutic alternative for a subgroup of patients with drug refractory, symptomatic, long lasting, and infrequent episodes of AF. Further efforts must be undertaken to reduce the patient discomfort associated with internal atrial defibrillation in an attempt to make this new therapy acceptable to a larger patient population with AF. (ABSTRACT TRUNCATED) PMID- 10810805 TI - [12th spring meeting of the Japanese Society of Allergology. Fukuoka, Japan. April 20-22, 2000. Abstracts]. PMID- 10810806 TI - [Regional meeting of the Japanese Association for Thoracic Surgery. Japan. 1999. Abstracts]. PMID- 10810807 TI - [Abstracts of the 105th annual meeting of the Japanese Association of Anatomists. Yokohama, Japan. March 29-31, 2000]. PMID- 10810808 TI - [The 73rd general meeting of the Japanese Leprosy Association. Kagoshima, Japan. March 9-11, 2000. Abstracts]. PMID- 10810810 TI - [The 70th annual meeting of the Japanese Society for Hygiene. Osaka, Japan. March 28-30, 2000. Abstracts]. PMID- 10810809 TI - [The 40th annual meeting of the Japanese Respiratory Society. Hiroshima, Japan. March 22-24, 2000. Abstracts]. PMID- 10810811 TI - [The 104th annual meeting of the Ophthalmological Society. Kyoto, Japan. April 6 8, 2000. Abstracts]. PMID- 10810812 TI - [The 84th Congress of the Japanese Society of Legal medicine. Wakayama, Japan. April 19-22, 2000. Abstracts]. PMID- 10810813 TI - [Cancer surgery of the pelvic basin. The anatomical controversies]. PMID- 10810814 TI - [The diagnostic activity within the compass of a day hospital surgical unit: a proposed organizational model]. AB - The authors report their own experience about the activity in a surgical day hospital. Day surgery should play a main role; nevertheless, some resources could be used in a diagnostic activity, to improve the turn-over of patients and minimize the time of charge in the surgical units. PMID- 10810815 TI - [Trends in stomach carcinoma. A statistical analysis of 1204 cases]. AB - The authors report a retrospective analysis of their experience of gastric cancer. One thousand two hundred and four patients, observed between 1977 and 1994, of whom 1,094 underwent surgery, were studied. Data analysis was performed, dividing this period as follows: 1977-1982, 1983-1988, 1989-1994. The results reveal that, over time, substantial changes occurred both in tumor characteristics and surgical approach. With regards to tumor characteristics, the disease weighs heavily, even if it is less frequent than in the past (25% reduction), striking younger subjects and presenting in increasingly more aggressive forms [higher frequency of proximal (+11.4%) and diffuse forms (+18.1%), reduction of advanced forms (-11.6%) not proportional to the increase of early forms (+64.5%), as well as increase of cases with a short clinical history (+15.2%)]. In terms of surgical approach, while the use of conservative surgery was reduced during the study period, extensive procedures were more widely used, particularly in the curative treatment of advanced tumors (subtotal gastrectomies: reduction of 39.5%; total gastrectomies: increase of 106%). The increasing use of extended surgery is linked to the modifications occurred in tumor characteristics and in pre and post-operative management rather then to changes in surgical approach. It must be noted, however, that such technical advances have not yielded an improvement in survival (p = n.s.); as such, it is likely that gastric cancer has became a more aggressive disease and, therefore, has counterbalanced the benefits of early diagnosis and extended exeresis. PMID- 10810816 TI - [Richter's hernia: a clinical case and the clinico-therapeutic considerations]. AB - The authors report a case of Richter's hernia. They underline main clinical and therapeutic patterns, emphasizing the need of an early diagnosis and surgery. This is a hernia of abdominal wall with partial entrapment of bowel wall (antimesenteric site) through a small ring. The incidence increased in the last years because of diffusion of laparoscopic techniques. Richter's hernia could be asymptomatic for a long time or show vanish sign. Sometimes this hernia can be diagnosed during surgery. The clinical signs are conclamated if hernia is complicated by strangulation. High mortality is justified by performing too late diagnosis and operation. PMID- 10810817 TI - [A case of carcinoid with multiple sites in the jejunum and distal ileum]. AB - Multiple carcinoid tumors of the small bowel with more than 3 lesions are very unusual. The authors report a case with 4 lesions, 2 of those localized in the jejunum with more advanced infiltration of the wall and extension to regional mesenteric lymph nodes, revealed by ultrasonography. The relative low incidence and particularly the vague, nonspecific clinical presentation, the unusual site in the jejunum, and failure of the radiological examine of one year before lead to not suspect this condition prior to US examination. However, the feature of asymmetric, concentric thickening of the bowel wall requiring a more accurate exam by CT with oral contrast was able to confirm the suspect of the intestinal tumor. The patient, 80 year old, underwent radical surgery with a wide lymph nodes dissection as well as double resection of the jejunum and distal ileum. The post-surgical outcome was uneventful. A 12-month follow-up is free of the disease. PMID- 10810818 TI - [Stenotic endometriosis of the sigmoid. Apropos a case]. AB - The authors report a rare case of stenotic sigmoid endometriosis which presented with constipation and dysmenorrhea in a 48-year-old woman without past gynecological history. There were no typical radiological and endoscopic findings and only pathological examination revealed endometriosis. The patient did well after sigmoid resection and bilateral salpingo-oophorectomy and has had no further complaints. The authors stress the rare location of the disease and review the literature about etiopathogenetic hypotheses and pathological features. The diagnostic value of clinical examination, barium enema and colonoscopy is low as endometriosis rarely involves the mucosa. This case led the authors to discuss about diagnostic difficulties, differential diagnosis and therapeutic options. PMID- 10810819 TI - [Pilomatrixoma or Malherbe's calcifying epithelioma. A case report and review of the literature]. AB - The case of a upper extremity calcifying epithelioma of Malherbe in a 16-year-old boy is reported. Clinical and pathological features of this extremely rare tumour, arisen from hair cortex cells, are reviewed. Pilomatrixoma has been reported not only as a benign lesion, or as a low-grade malignant lesion with a tendency to recur locally, but also as a highly malignant tumour. Factors influencing clinical differential diagnosis and prognosis are discussed as well. PMID- 10810820 TI - [Spontaneous biliodigestive fistulae. The clinical considerations, surgical treatment and complications]. AB - To reevaluate the current feature of spontaneous bilioenteric fistula we reviewed 81 cases who had been treated for biliary fistula between 1948 and 1998. After a review of the literature on this subject, the multiple problems relate to pathological anatomy, pathogenesis and physiopathology are discussed. Of 81 patients, 55 were women and 26 were men with the average age of 54.5 years. The most common type of fistula was cholecysto-duodenal (55 cases--68%), followed by cholecysto-colonic (11 cases--13.6%), choledocho-duodenal (7 cases--8.6%), cholecysto-gastric (4 cases--4.9%) and duodeno-left hepatic duct fistula (4 cases). The authors have found in 41 cases the gallstone ileus complications, in 12 cases inflammatory disease of biliary three, in 8 cases hemobilia, gallstone ileus with perforation and digestive hemorrhage compliances respectively. All the patients were treated with surgery. A first procedure consists of enterolithotomy, in gallstone ileus cases, followed by biliary surgery. In 14 patient the general or local conditions argued against one-stage procedure and two-stage procedure had been considered. In 63 patients a cholecystectomy was done, 15 were treated with enterolithotomy and 8 with intestinal resection. Seven patients with gastroduodenal ulcer based fistula have required a gastroduodenal resection. The mortality was 13.6% (11 cases). PMID- 10810821 TI - [Biliary cystadenocarcinoma. A case report]. AB - The authors report a case of cystadenocarcinoma of the biliary tract, an uncommon tumor of the liver. A 68 aged white Caucasian jaundiced male subwent intervention for a tumor of the 4th hepatic segment. Forty-two months after discharge he is still alive without any secondary lesion. On the 16th follow-up month a prostatic malignancy was diagnosed. Differential diagnosis with other cystic lesion is an exacting and difficult target. Usual techniques such as US, CT, NMR infrequently get a correct diagnosis. Often only in surgical room a real diagnosis is achieved. In the aim to avoid tumor relapse the best surgical procedure is a large hepatic resection. PMID- 10810822 TI - [The role of endoscopic sphincterotomy in the therapy of choledocholithiasis: a review of our experience]. AB - The treatment of common bile duct stones is nowadays endoscopic sphincterotomy. The authors reviewed their series from 1992 to 1999 and confirm that this is a safe, definitive, with low complication rate treatment if is performed in specialized centers and reduces the duration of hospitalization. Operative ERCP is successful is about 96-97% of the cases. There was no mortality related to the technique. PMID- 10810823 TI - [Inhaled foreign bodies: first aid]. AB - In the last 30 years 1,484,576 patients with traumatic lesions of different kind of seriousness were observed and managed. By mean of retrospective investigation, the authors selected and statistically analysed a sample of 95 patients treated for accidental tracheobronchial foreign bodies inhalation: authors' aim was to punctually determinate--step by step--diagnosis and treatment procedure in a "time-saving" frame. Inhalation of foreign bodies is a very dangerous event, particularly in children. In authors' experience, in 60% of cases they were children under 3 years; in 25% of cases children under 10; in 10% patients from 10 to 60 and in 4.5% over 60. As regards the gender of the patients, the male/female ratio has been 2 to 1 in favour of male. Since 85% of the study population was represented by children under 10, its clear that male children in this range of age has a greater statistical risk for accidental tracheobronchial foreign bodies inhalation. PMID- 10810824 TI - [The usefulness of lateral internal sphincterotomy combined with hemorrhoidectomy in the treatment of hemorrhoids: a randomized prospective study]. AB - The aim of this prospective randomized study was to investigate anorectal manometric findings in hemorrhoid patients and to evaluate the clinical benefits and physiological consequences of additioning a lateral internal sphincterotomy (LIS) to haemorrhoidectomy. Anorectal manometry was preoperatively performed in forty-eight consecutive patients with prolapsed piles; resting and squeeze pressures, sphincter length and rectoanal inhibitory reflex were recorded. Ten healthy volunteers served as controls. Six patients were excluded because no raised and pressures were found. Forty-two patients were randomised: Group 1 (n = 22) patients underwent haemorrhoidectomy plus LIS; Group 2 (n = 20) patients underwent haemorrhoidectomy alone. Postoperative course was carefully evaluated; all patients were questioned about continence and anorectal manometry was repeated twice. Sphincter anomalies were found in 87.5% of patients. Haemorrhoidectomy alone did not affect anal pressures, which returned into the normal ranges after sphincterotomy. Postoperative course was better in LIS group. Anal stricture was seen in four patients without sphincterotomy; no patients with LIS experienced and incontinence. This study shows that high and pressures are very frequent in hemorrhoid patients; they are not due to hypertensive and cushions and might have a pathogenetic role. Anorectal manometry is very useful to identify patients with raised anal pressures; in these cases additioning a lateral internal sphincterotomy to haemorrhoidectomy seems justified; it significantly improves postoperative course and can be safely performed. PMID- 10810825 TI - [The endoscopic treatment of foreign bodies in the tracheobronchial tree]. AB - Details are presented on the endoscopic treatment of a series of 70 patients with tracheal on bronchial foreign bodies. Diagnosis was made on the basis of a careful examination of anamnestic, clinical, radiological and endoscopic data. PMID- 10810826 TI - [Axillary lymphadenectomy and its complications]. AB - Local and regional complications of axillary dissection are evaluated. According to other authors, the most frequent ones are infections, seroma and lymphoedema. Sentinel node biopsy can reduce the number of dissections and their complications. PMID- 10810827 TI - Reducing stress among pediatric oncology nurses. PMID- 10810828 TI - Testing the stress-response sequence in pediatric oncology nursing. AB - This article summarizes six studies completed at St. Jude Children's Research Hospital on role-related stress experienced by pediatric oncology nurses. Two models, the Environmental Care model and the Stress-Response Sequence (SRS) model, provided the theoretical guidance for these studies. The studies included initial efforts to identify and define role-related stressors, reactions, and consequences; to develop a psychometrically sound instrument to measure role related stressors; and to test portions of the SRS model. Subsequent studies included a complete testing of the SRS model, an exploratory effort to define role-related meaning, and a testing of the revised SRS model. Findings to date indicate that pediatric oncology nurses experience moderate to high role-related stressors but report notably positive consequences, including high job satisfaction and group cohesion and low intent to leave the setting or the specialty. High levels of role-related meaning seem to sustain the commitment of pediatric oncology nurses in their efforts to deal with role-related stressors. The current study will help to determine if the revised SRS model adequately explains the role-related stress response of pediatric oncology nurses. If it does, the revised model will be used to develop and test interventions designed to increase role-related meaning and effective coping reactions. PMID- 10810829 TI - Historical development and past use of the stress-response sequence model in healthcare. PMID- 10810830 TI - The stress-response sequence model of stress in pediatric oncology nurses. PMID- 10810831 TI - Problem solving skills training in pediatric oncology nurses: mediating the stress-response sequence. PMID- 10810832 TI - Using the stress-response sequence model in pediatric oncology nursing: implications for practice. PMID- 10810833 TI - Studying the stress-response sequence in pediatric oncology nursing. School of Nursing, Northeastern University. PMID- 10810834 TI - Identifying nursing research priorities in a pediatric clinical trials cooperative group: the Pediatric Oncology Group experience. AB - If nurses involved in pediatric oncology clinical trials are to use resources and manpower most effectively, nursing research priorities must be set that meet the nursing care issues of nurses at the bedside. Using a Delphi technique, nurses from the Pediatric Oncology Group (POG) were surveyed in Round 1 to discover patient care concerns or other issues in pediatric oncology nursing related to clinical trials. The 87 research ideas generated were reviewed by a panel of nurse experts, and a total of 57 unique research topics were delineated. These topics were then used for Round 2 of the Delphi study, which identified the top 10 nursing research priorities of the POG nurses participating in the study. PMID- 10810835 TI - Expectations. Pediatric oncology nursing. PMID- 10810836 TI - The "hired gun" effect: assessing the effect of pay, frequency of testifying, and credentials on the perception of expert testimony. AB - Three experiments addressed the proposition that jurors use short cuts in processing information when confronted with expert scientific testimony. The results of the first two studies demonstrated that experts who are highly paid for their testimony and who testify frequently are perceived as "hired guns." They are neither liked nor believed. The results of the third experiment replicated the hired gun effect and showed that it is most likely to occur when the testimony is complex and cannot be easily processed. The results were discussed in terms of the theoretical differences between central and peripheral processing of persuasive messages in a legal context. PMID- 10810837 TI - Studying perceptions of juror influence in vivo: a social relations analysis. AB - Because of legal constraints and statistical limitations there has been little research on social influence in actual juries. We used Kenny's (1994) social relations model to examine jurors' perceptions of social influence in the jury. After rendering a verdict in criminal or civil court cases, jurors rated how influential each member of the jury had been and provided self-reports of their personality traits. Perceptions of influence in the jury were mostly in the eye of the beholder, with jurors high in Conscientiousness and low in Openness being most likely to report that they were personally influenced by other jurors. There were small but statistically significant levels of consensus in the ratings of how influential the jurors were. To the extent that they did agree, jurors rated extraverted, tall men as most influential. PMID- 10810838 TI - Compensating plaintiffs and punishing defendants: is bifurcation necessary? AB - Critics of the civil jury have proposed several procedural reforms to address the concern that damage awards are capricious and unpredictable. One such reform is the bifurcation or separation of various phases of a trial that involves multiple claims for damages. The purpose of this study was to assess the effects of bifurcating the compensatory and punitive damages phases of a civil tort trial. We manipulated the wealth of the defendant and the reprehensibility of the defendant's conduct (both sets of evidence theoretically related to punitive but not to compensatory damages) across three cases in a jury analog study. We wondered whether jurors would misuse the punitive damages evidence in fixing compensatory damages and whether bifurcation would effectively undo this practice. Our findings indicated that mock jurors did not improperly consider punitive damages evidence in their decisions about compensation. Moreover, bifurcation had the unexpected effect of augmenting punitive damage awards. These findings raise questions about the merits of bifurcation in cases that involve multiple claims for damages. PMID- 10810839 TI - Effects of judicial instructions and case characteristics in a mock jury trial of battered women who kill. AB - This study examined the effects of judicial instructions on the outcome of a mock jury trial that involved a woman who pleaded self-defense after killing her abusive spouse. Jurors were instructed to adopt either an objective or a subjective standard of reasonableness when reaching a verdict. Within objective/subjective instruction conditions, half of the juries viewed a case in which the woman killed her abuser while he was attacking her (confrontational) and the remaining half viewed a case in which she killed him while he was asleep (no confrontation). Juries in the subjective conditions returned significantly more not guilty verdicts than jurors in the objective conditions. At the individual juror level, participants hearing subjective instructions were significantly more likely to rate the defendant as not guilty than jurors given objective instructions when the abuse was nonconfrontational. PMID- 10810840 TI - Selecting foils for identification lineups: matching suspects or descriptions? AB - Two experiments directly compare two methods of selecting foils for identification lineups. The suspect-matched method selects foils based on their match to the suspect, whereas the description-matched method selects foils based on their match to the witness's description of the perpetrator. Theoretical analyses and previous results predict an advantage for description-matched lineups both in terms of correctly identifying the perpetrator and minimizing false identification of innocent suspects. The advantage for description-matched lineups should be particularly pronounced if the foils selected in suspect matched lineups are too similar to the suspect. In Experiment 1, the lineups were created by trained police officers, and in Experiment 2, the lineups were constructed by undergraduate college students. The results of both experiments showed higher suspect-to-foil similarity for suspect-matched lineups than for description-matched lineups. However, neither experiment showed a difference in correct or false identification rates. Both experiments did, however, show that there may be an advantage for suspect-matched lineups in terms of no-pick and rejection responses. From these results, the endorsement of one method over the other seems premature. PMID- 10810841 TI - The bottom line: the effect of written expert witness statements on juror verdicts and information processing. AB - Mock jurors recruited from jury rolls were either not given written statements of expert witnesses' direct testimony or were provided with such statements before or after the presentation of that testimony. Presentation of the statements before the testimony and cross-examination provided jurors with a schema that allowed them to distinguish more effectively among the claims of four differentially worthy plaintiffs because they processed more probative evidence than other jurors. Jurors in receipt of written statements before the testimony found the evidence to be more comprehensible than other jurors. Jurors provided with written statements following testimony and cross-examination were able to differentiate between the most and least severely injured plaintiffs, whereas jurors not in receipt of any written statements were unable to differentiate among any of the differentially worthy claimants. The limitations of this case management technique and of the study are discussed. PMID- 10810842 TI - Is that your final answer? PMID- 10810843 TI - Should childhood immunizations be given when children have a pediatric visit to the emergency department? PMID- 10810844 TI - "It wasn't working. " Women's experiences with short-term breastfeeding. AB - PURPOSE: To investigate the lived experiences of women who initiate breastfeeding but stop within the first 2 weeks after birth. DESIGN: Phenomenology. METHODS: Interviews were recorded on audio tape, transcribed verbatim, and analyzed for themes by an interdisciplinary group of researchers. RESULTS: Women described a clash or incongruity between highly idealized expectations and early breastfeeding problems. This led to incremental disillusionment and cessation of breastfeeding attempts. Women spoke poignantly about a sense of failure, guilt, or shame and had lingering self-doubts about not continuing with breastfeeding. CLINICAL IMPLICATIONS: Women need more accurate information about the process of initiating breastfeeding. Nurses should avoid teaching breastfeeding practices that can confuse infants and sabotage mothers and should work toward providing sensitive care that respects personal boundaries of mothers. Additionally, provisions need to be made for continued support and consultation after the mother's discharge from the hospital or birthing center. Finally, nurses and other health professionals should understand that women who stop breastfeeding at an early date may feel guilty about doing so, and may need assistance in resolving feelings about having made that choice. PMID- 10810845 TI - Mothers' perceptions of parenting children with disabilities. AB - PURPOSE: To describe how mothers of children with physical disabilities identified their parenting strengths and weaknesses, encouraged their child's social skills and learning, nurtured and disciplined their child, and perceived their parenting successes and problems. DESIGN: Grounded theory. METHODS: Focused telephone interviews were conducted with mothers of preschool to elementary schoolage children with mild to moderate physical disabilities. The mothers were asked what they were doing well and "not well" in parenting their disabled child, their difficulties, their child's preparation for other children's questions/teasing, their encouragement of social skills and learning, and their discipline and nurturance. RESULTS: The theory that emerged from the data was parental straddling, which occurred on three levels. Parents were living in the past and the present, striving to view their child as "normal" when, in fact, the child was disabled, and were simultaneously dealing with their own and their child's issues and feelings. CLINICAL IMPLICATIONS: Nurses need to understand the complexity of the parents' straddling their roles and tasks. They can provide needed emotional support by allowing parents to express their fears and feelings in an atmosphere of nonjudgmental acceptance. Nurses can prepare parents for anticipated grief work, reassure them that their experiences are expected and normal, reinforce their use of normalization strategies, and help them separate their own from their child's issues and feelings. PMID- 10810846 TI - Expectant fathers and labor epidurals. AB - PURPOSE: A qualitative research study was conducted to describe and explain expectant father's experiences during labor when their partners receive labor epidurals. DESIGN: Grounded theory. METHODS: Seventeen couples were interviewed using a semi-structured interview format. RESULTS: Two major concepts were identified: "Losing Her" and "She's Back." The men reported that these were the two critical points at which the epidural affected their experience. The theory, "Cruising through Labor," explained the fathers' experiences. CLINICAL IMPLICATIONS: Childbirth preparation classes that describe and explain women's responses to the increasing pain of labor contractions, the exhaustion of labor, and the effect of epidural analgesia could possibly assist men in preparing for the changes they will witness in their partners during the labor experience. Knowing that these changes are normal responses to the effects of labor might reduce men's levels of anxiety, frustration, and sense of helplessness. The labor nurse is a major support to the expectant fathers. Labor nurses who remain in the room during this phase, explain what is happening, and include the expectant father in their nursing care provide the best support. PMID- 10810847 TI - Enhancing parent-child interaction with a prenatal couple intervention. AB - PURPOSE: To determine the effect of a prenatal couple group intervention on parent-child interaction postbirth. DESIGN: Quasiexperimental study. METHOD: A nonrandomized convenience sample of treatment group (TG) couples (n = 35) who attended an additional prenatal three-class series was compared to a control group (CG) from childbirth education classes on measures of videotaped parent child interaction using the NCATS tool. The intervention class series was based on individual and couple changes in meaning/identity, roles, and relationship/interaction during the transition to parenthood. It addressed mother/father roles, infant communication abilities, and patterns of the first 3 months of life in a mutually enjoyable, possibility-focused way. RESULTS: T-tests and ANCOVA on NCATS scores between groups showed higher TG scores for mothers in sensitivity to cues, for fathers in social-emotional growth fostering, and for couple mean scores in social-emotional growth fostering, couple mean response to child distress, caregiver total, and caregiver-child total. Higher contingency scores were also found in the TG group. Fewer TG mothers and fathers fell below NCATS lower cutoff scores. CLINICAL IMPLICATIONS: Interventions that enhance mutual parent-child interaction through increased sensitivity to cues and responsiveness to infant needs or signals are important avenues for facilitating secure attachment, father and mother involvement, optimal development, and prevention of child abuse and neglect. The positive approach to this intervention invites couples to see themselves as developing with their infants over time, and to view their infants in new ways that will help develop satisfying, self reinforcing patterns of interaction. PMID- 10810848 TI - A description of birth in Finland. AB - The health care of women and children living in Finland is described. In the Finnish health care delivery system, perinatal health care is family centered, maximizing professional support through use of nurses in the delivery of primary care across the childbearing year and technological interventions are judiciously used based on evidence of efficacy. PMID- 10810849 TI - Public health nursing data. Building the knowledge base for high-risk prenatal clients. AB - PURPOSE: To examine and compare referral reason, clinical pathway variance, and intensity of need for care for high-risk prenatal clients of public health nurses. DESIGN: A prospective, descriptive design was used for this pilot study. METHODS: Reasons for referral to the high-risk prenatal home visitation program, variances from a prenatal clinical pathway, and intensity of need for care scores obtained using the Community Health Intensity Rating Scale (CHIRS) were collected at 28 weeks and 38 weeks of gestation from the clinical records of 20 high-risk prenatal clients (age range 16-43 years) visited by five expert public health nurses in one midplains public health nursing department. RESULTS: Findings indicated that the three sources of clinical data provided congruent but not identical data, with each contributing elements needed to specify public health nurses interventions for high-risk prenatal clients. CLINICAL IMPLICATIONS: Collaborative clinical research studies such as this one are useful in advancing evidenced-based practice in clinical agencies. The congruence between the clinical path variance and the intensity of need for care scores reaffirms the importance of the domain of health behavior intervention as a major aspect of public health nurses practice. PMID- 10810850 TI - Alcohol use and adolescent pregnancy. AB - Reports indicate that 33.4% of adolescents engage in heavy episodic alcohol consumption, and that 34.8% of adolescents are sexually active by the age of 15 without using any form of contraception. Combined, these activities can lead to adolescent pregnancy complicated by fetal alcohol syndrome (FAS), a rare disorder that is totally preventable by abstinence from alcohol during pregnancy. This article offers some primary prevention strategies that nurses can adopt in working with adolescents and communities to help prevent FAS. PMID- 10810851 TI - Informatics in nursing practice. PMID- 10810852 TI - Informed consent for adult patients: Part two. PMID- 10810853 TI - Barriers to prenatal care. PMID- 10810854 TI - Laboring down. PMID- 10810855 TI - Infant sleep position PMID- 10810856 TI - Infant sleep position PMID- 10810857 TI - [When the brain is in acute danger. Are you fit for the neuro-emergency?]. PMID- 10810858 TI - [Tips for first aid for comatose patients. Brief assessment points the way. Interview by Dr. Beate Schumacher]. PMID- 10810859 TI - [In stroke, every minute counts. Proper positioning, intravenous access, measuring O2, heart rate--admission to the clinic!]. PMID- 10810860 TI - [Testosterone--sometimes too much, sometimes too little. The man--an eternal patient?]. PMID- 10810861 TI - [New flu therapy. Progress or burden for the budget?]. PMID- 10810862 TI - [New approach to chronic inflammatory bowel diseases. Worm ova by prescription?. Interview by Petra Eiden]. PMID- 10810863 TI - [Removing adenomas reduces colon carcinoma risk up to 90%. Effective cancer prevention with the endoscope]. AB - Endoscopic polypectomy is considered the method of choice for the diagnosis and treatment of colorectal polyps. 70-80% of all colorectal polyps are adenomas, which are precursors of colorectal carcinoma. The endoscopic detection and rigorous removal of colorectal polyps is an effective means of preventing colorectal carcinoma, since the incidence of the latter can be reduced by 90%. If technically possible, a snare should be used to remove all polyps larger than 5 mm in diameter. Tiny polyps may be removed with forceps. The removed polyp must be send for histological work-up and the results obtained used to plan the further steps or post-polypectomy surveillance. In defined cases (removal with adequate clearance, well-differentiated carcinoma, low risk situation), endoscopic polypectomy of adenomas with invasive carcinoma (pT1 carcinoma) is now the only treatment needed. Further surgical resection is not necessary. Surveillance is as for surgically removed colorectal carcinoma. PMID- 10810864 TI - [Can colorectal carcinomas be resected laparoscopically? Technically possible, long-term outcome is still uncertain]. AB - DESIGN: The efficacy of laparoscopic resection of colorectal tumours was evaluated in a prospective observational study. PATIENTS: All patients with elective laparoscopic resection for colorectal tumours between 1995 and 1997 were included. MAIN RESULTS: Laparoscopic resections were accomplished in 70 patients with a mean age of 61.9 +/- 15.6 years--most of them (n = 45) left-sided. The operative time was 232 +/- 75 minutes and the estimated blood loss 235 +/- 123 ml. In six patients (8.6%) laparoscopic-assisted procedures were performed, and no patient had to be converted to a conventional midline incision. Intraoperative complications occurred in one patient, postoperative general complications in 2.9% and surgical complications in 5.7%. Clinically relevant anastomotic leaks were not recorded. No patient died postoperatively. Reconvalescence was short, first bowel movement was documented after 3.3 +/- 1.3 days. Postoperative hospital stay was 10.1 +/- 3.1 days. CONCLUSION: Laparoscopic resection of colorectal tumours can be accomplished with a low complication rate with proper selection of patients and experience of the surgical team. However, the indication for laparoscopic resection must consider all aspects of an oncological treatment. PMID- 10810865 TI - [Noma in Africa. The face of poverty]. PMID- 10810866 TI - [Management of chronic tinnitus. Counseling--therapeutic apparatus- psychotherapy]. PMID- 10810867 TI - [Thrombocyte control after heparin administration is essential. Thrombosis ABC, 12: Heparin-induced thrombocytopenia]. PMID- 10810868 TI - [Ulcus cruris. 1: Etiology, prevalence and diagnosis]. PMID- 10810869 TI - [Painless ulcer of the penis. Granuloma inguinale tropicum]. PMID- 10810871 TI - [5. Fever--a symptom of many etiologies)]. PMID- 10810870 TI - [Montgomery confronts KVB. If money follows performance, there's less for established physicians. Interview by Werner Loosen]. PMID- 10810872 TI - [Congenital thrombophilia caused by protein C deficiency]. AB - To clarify the prevalence of protein C deficiency in patients with cardiovascular problems, we identified 54 patients with protein C deficiency by screening approximately 26,800 patients. The observed prevalence of protein C deficiency was 1 in 500. From the results of genetic analysis of 67 Japanese families with protein C deficiency, the recurrent defects of Phe139Val, Arg169Trp, Val297Met, Met364Ile substitutions and a G8857 deletion were accounting 49% of Japanese families with protein C deficiency. The defects including Phe139Val and Met364Ile substitutions and a G8857 deletion were only found in Japanese subjects and seemed to be a founder effect. Protein C deficiency is associated with a predisposition to venous thrombosis. It is not clear whether the deficiency is involved in arterial occlusion. We identified 34 patients with protein C deficiency manifested 45 episodes of arterial occlusive disease. We compared the onset age of arterial occlusive disease between patients with protein C deficiency and a group of patients with normal protein C levels. We found the early onset of acute myocardial infarction and atherothrombotic cerebral infarction in patients with protein C deficiency. Thus, our study suggests that congenital protein C deficiency contributes to earlier onset of arterial occlusive diseases in Japanese subjects. PMID- 10810873 TI - [Antiphospholipid antibodies and thrombosis: the putative mechanisms of hypercoagulable state in patients with anticardiolipin antibody]. AB - Antiphospholipid antibodies are well recognized as associated with serious clinical complications such as arterial and venous thrombosis and recurrent spontaneous abortion. These complications are collectively called antiphospholipid syndrome(APS). The mechanisms responsible for the thrombosis are unclear. We reported three mechanisms. beta 2-glycoprotein I(beta 2GPI) inhibited activated protein C(APC) activity and, furthermore, APC activity decreased by the addition of monoclonal aCL and beta 2GPI. Monoclonal anticardiolipin antibodies(aCL) seemed to enhance the inhibition of APC procoagulant activity caused by beta 2GPI. Monoclonal aCL in the presence of beta 2GPI also increased the activity of plasminogen activator inhibitor(PAI)-1 in the mixture of tissue plasminogen activator(t-PA) and PAI-1 by inhibiting the function of beta 2GPI, which increased the remaining t-PA activity in the mixture. The formation of thrombin-antithrombin complexes(TAT) in APS was impaired. The level of TAT in APS did not increase, however the level of prothrombin fragment 1 + 2 (F1 + 2) increased. Therefore, free thrombin present in patients' blood may contribute to thrombosis in APS. These reports indicate that thrombosis in APS may be caused by several thrombogenic factors that stimulate aCL. PMID- 10810874 TI - [Application of hemostatic molecular markers for diagnosis of thrombosis]. AB - We examined various hemostatic molecular markers in patients with disseminated intravascular coagulation(DIC), deep vein thrombosis(DVT), pulmonary embolism(PE), acute myocardial infarction(AMI), cerebral thrombosis(CT) and thrombotic thrombocytopenic purpura(TTP). Global tests were sensitive for DIC but not for pre-DIC. However, hemostatic molecular markers such as soluble fibrin were sensitive for both DIC and pre-DIC. Hemostatic molecular markers were also useful for analysis of DIC in a baboon DIC model. Activated protein C-protein C inhibitor complex and plasminogen activator inhibitor-I were useful for the diagnosis of DVT, PE, AMI or CT. These findings suggests that hemostatic molecular markers are useful for the diagnosis of various thrombotic disorders. PMID- 10810875 TI - [Endothelial cells and coagulation abnormalities]. AB - Endothelial cells have two important anticoagulant systems, heparan sulfate antithrombin system and thrombomodulin-protein C system. Under physiological conditions, these two systems inhibit activation of coagulation on endothelial cells. However, under inflammatory conditions, tumor necrosis factor(TNF)-alpha or other cytokines produced by monocytes reduce the anticoagulant properties of endothelial cell by downregulating expression of heparan sulfate and thrombomodulin on endothelial cells. Antithrombin stimulates prostacyclin generation from endothelial cells by interacting with heparan sulfate of endothelial cells and generated prostacyclin inhibits TNF-alpha production by monocytes. Activated protein C inhibits TNF-alpha production by monocyte dependent of its protease activity. Thus, antithrombin and activated protein C might inhibit the endothelial perturbation induced by cytokines. Antithrombin regulates TNF-alpha induced tissue factor expression on endothelial cells by an unknown mechanism. Thus, antithrombin and activated protein C might be useful agents for treating coagulation abnormalities associated with sepsis or other inflammation because these agents inhibit not only coagulation but also downregulation of anticoagulant activities of endothelial cells. PMID- 10810876 TI - [Assay principles of antiphospholipid antibodies and heterogeneity of the antibodies]. AB - The antiphospholipid syndrome(APS) is characterized by predominant clinical features of venous and arterial thrombosis and recurrent pregnancy loss accompanied by antiphospholipid antibodies(aPL) such as anticardiolipin antibodies(aCL) and lupus anticoagulant(LA). In 1990, three individual research groups, including us, first reported that a 50 kD plasma cofactor is required for the binding of aCL to cardiolipin(CL) and now, beta 2-glycoprotein I(beta 2-GPI), which binds to anionic phospholipids(PLs), is widely believed to be the major antigen for aCL. It was also reported that epitopes for such aCL are cryptic and that they appear only when beta 2-GPI interacts with lipid membranes containing anionic PLs, such as CL and phosphatidylserine, or with a polyoxygenated polystyrene surface. In contrast, prothrombin was recently identified as the "true" antigen for LA. In this review paper, we would like to describe on specificity of aPL and also on a possible mechanism on autoantibody-dependent development of atherosclerosis. PMID- 10810877 TI - [Antiphospholipid antibody syndrome in adverse pregnancy]. AB - As a single test beta 2-glycoprotein I-dependent anticardiolipin antibody(beta 2GPI-dependent aCL) appears to be superior to cofactor-independent anticardiolipin antibody or any other single conventional antiphospholipid antibody for the detection of autoantibody-associated conditions of reproductive failure. beta 2GPI-dependent aCL are significantly highly associated with adverse pregnancy outcomes in healthy pregnant women and can be used for prediction, whereas beta 2GPI-independent aCL can not. Anticardiolipin antibodies and Lupus anticoagulant(LAC) define two distinct but partly related populations. LAC and anti beta 2GPI antibodies appear to be associated with pregnancy loss, with LAC being linked not only to spontaneous abortions in the first trimester but also to miscarriages in the second trimester. The live birth rate in patients strongly positive for antiphospholipid antibody(aPL) is lower than that in patients with moderate aPL production even if treatment with prednisolone, heparin, high-dose immunoglobulin, and/or low-dose aspirin is performed during pregnancy. However, low-dose aspirin is useful to treat cases of moderate aPL so that distinction between the two groups is warranted. PMID- 10810878 TI - [Transplantation for amyloidosis]. PMID- 10810879 TI - [Interference by D-mannitol on serum D-arabinitol determination by enzymatic assay]. AB - We examined a disparity in D-arabinitol values between two commercial assay kits, LABOFIT and ARABINITEC-AUTO. The determined values by the former were increased by 26%(y = 0.2643x) because of concomitant D-mannitol, whereas those by our newly developed ARABINITEC-AUTO was increased only by 2%(y = 0.0242x). Of 109 samples, 5 samples were found to contain more than 100 mumol/l of D-mannitol. A clear relation(r = 0.89) was noted between the degree of disparity between measurements by the two methods and D-mannitol concentrations in samples. Thus, we have proved that the disparity is mainly caused by D-mannitol. PMID- 10810880 TI - [Expression of reactive oxygen-species related enzymes in endothelial cells stimulated with glycated lipoproteins]. AB - Glycated lipoproteins, which elevate the blood in diabetic patients, cause direct attenuation of endothelial function. Increased glycation of apolipoproteins may play a trigger role in the accelerated development of atherosclerosis in the patient with diabetes. Here we assessed whether glycated lipoproteins affect on the endothelial function with particular emphasis on the role of reactive oxygen species in vitro. Incubation of human aortic endothelial cells(HAEC) with glycated LDL had little influence on the expression of antioxidant enzymes such as nitric oxide synthase(NOS), Cu2+Zn(2+)-superoxide dismutase (Cu2+Zn(2+)-SOD), catalase, and p22 phox in the cells. In contrast, exposure of glycated HDL induced a marked decrease of Cu2+Zn(2+)-SOD, catalase, and endothelial NOS as well as a slight increase of p22 phox in HAEC in term of both protein and mRNA expression, suggesting that increased formation of reactive oxygen species such as O2- and OH radical participate in the deterioration for the function of vascular endothelial cells in diabetic patients. PMID- 10810881 TI - [Genome analysis of Helicobacter pylori by pulsed-field gel electrophoresis]. AB - The molecular epidemiology of total 121 isolates of Helicobacter pylori was analyzed by pulsed-field gel electrophoresis method with restriction enzyme Spe I. Seventy-seven isolates were separated from the clinical samples, 36 isolates from pyloric antrum and the body of stomach of 18 patients and 8 isolates from pyloric antrum of 4 patients that include one colony before and after sterilizing treatment to each patient. Seventy-five in 77 isolates showed different genomic types respectively, and the other 2 isolates had the same genomic type and were suspected to be caused by intersective infection of medical workers or the instruments that used in examination because they were from patients who were examined by gastric microscope in same time and same laboratory. In isolates from 4 patients who were treated by sterilizing method, 2 patients showed same genomic types with that observed before the treatment, and one patient showed an incomplete treatment because the genomic type of its colony was is similar, and another patient could be infected again because its isolates showed different genomic type. In 18 patients whose isolates were separated from pyloric antrum and body of the stomach respectively to each person, isolates of 3 patients showed different genomic types in the two different part of stomach indicating that they had two and more clones of H. pylori. PMID- 10810882 TI - [Dissociative disorders: from Janet to DSM-IV]. AB - I reviewed the literature on dissociation and dissociative disorders from Pierre Janet to DSM-IV, and examined the current trends in research. Janet's theory on hysteria is multifaceted, and is based on three psychological models. Based on a hierarchical model, Janet related hysteric symptoms to the activities within the lower strata of mental hierarchy (automatisms psychologiques), which were demonstrably shown in somnambulism. A second model was based on the concept of a psychological system, which was hypothetically composed of ideas, images, feelings, sensations, and movements. According to this model, dissociation of psychological functions was fundamental to the mechanism of hysteria: loss of integration was thought to engender fixed ideas (ideas fixes) and to lead to the development of a system totally isolated from the whole personality system. Janet also attempted to explain various mental disorders using an economic model. He referred to a loss of equilibration between psychological force and psychological tension. Thus, an unexpected emotional experience was conceived to cause a consumption of reserved psychological force, which was in turn followed by exhaustion associated with hysteric symptoms. Whereas most current researchers regard Janet as the first to study psychological trauma as a principal cause of dissociation, I feel it is important to note that he also emphasized the role of stigmata, i.e., permanent traits of hysteric patients, which were represented as a suggestibility and a tendency toward a narrowing of the consciousness field. Discussion about dissociation and its relation to trauma all but disappeared after Janet. However, during the Second World War and post-war period, some psychiatrists began to pay attention to two emerging phenomena: a high incidence of dissociative symptoms such as fugue and amnesia among combatants, and traumatic neurosis frequently observed among ex-inmates of concentration camps. In the 1970s, interest in dissociation and trauma was revived in different areas: the feminism movement was linked with concerns about child sexual abuse, public curiosity about multiple personalities was heightened by novels and movies, and recognition of posttraumatic stress disorder (PTSD) among Vietnam War veterans. In 1980, dissociative disorders were finally adopted as a diagnostic category in the official nomenclature of DSM-III. Although current research on dissociation is being carried out in various fields, two basic assumptions, reflected in the definition of DSM-IV, can be made. One is the "trauma-genic hypothesis," and the other is the great importance attached to multiple personality disorder (MPD). According to the predominantly held view, dissociation represents a reaction to early traumatic experience, especially sexual and physical abuse in childhood. In contrast, some authors argue that the causality of childhood traumatic experience has not been empirically confirmed, and other factors such as the influence of the environment and the predisposition of patients should be taken into consideration. MPD, which was originally described as an unusual phenomenon in classical literature, is currently thought to be a common type of dissociation. However, the reported rapid increase in the number of MPD patients in North America may be partially due to over-diagnosis and inclusion of iatrogenic cases. Significance is also given to MPD in respect to classification of dissociative phenomena. According to the widely held scheme of a "dissociative continuum," which ranges from normal experiences such as daydreams to pathological states, MPD is placed at the extreme end of the continuum. Furthermore, most researchers tend to classify MPD as the severest dissociative disorder due to chronic trauma. On this point, there seems to be confusion about "extremity" and "severity" of MPD. I conclude that the trauma-genic hypothesis of dissociation and the overemphasis placed on MPD should be reexamin PMID- 10810884 TI - [Management of the mentally disturbed with criminal records in England]. PMID- 10810883 TI - [Current status and problems in the management of mental patients criminal records in Japan]. PMID- 10810885 TI - [Evaluation and treatment of the mentally ill with criminal records: a Calgary model]. PMID- 10810886 TI - [Current status of treatment of psychiatric patients with criminal records at public hospitals]. PMID- 10810887 TI - [Problems related to management of mentally disturbed with criminal records: an attorney's viewpoint]. PMID- 10810889 TI - [On the policy to manage the mentally disturbed with criminal records in Japan: a viewpoint by the Ministry of Health and Welfare]. PMID- 10810888 TI - [On the ideal policy for managing the mentally disturbed with criminal records in Japan--a viewpoint of the Ministry of Justice]. PMID- 10810890 TI - [On the policy for managing the mentally ill with criminal records in Japan: a viewpoint by the Japanese Legal Association]. PMID- 10810891 TI - [Religion and psychiatry in the modern society]. PMID- 10810892 TI - [Modern art therapy]. PMID- 10810893 TI - [The law and research meetings held by clinical psychiatry groups: the current status and future prospect]. PMID- 10810894 TI - [Phenomenological study on the process to onset and the structure of symptoms in eating disorders]. AB - Psychogenic experiences occurring prior to the onset of eating disorders had previously been ignored as not corresponding to the symptoms. Recently, we have been considering these experiences as outbreak conditions, and have done phenomenological analysis of the experiences themselves with the following results. We found that patients have experienced 1.) "shame", and following that, and from the phenomenological consideration of the experience of "shame", we have observed 2.) "alienation" occurring in each patients, leading to "isolation." In "isolation", the patient is driven to despair due to existential anxiety, and as the ultimate in despair, the patient shows the attitude of 3.) "One wishing to be oneself in despair." (Kierkegaard, S.), or "haughtiness" which is the attitude of "a person who understands himself or herself to be a free design, and considers others as 'existence-others' reached through this design." (Sartre, J. P.) Here, the patient shows "consumption" as a symptom in either of the forms of surfeit (bulimia) or the consumption of one's own body (anorexia): wasting. 4.) These are defined as "conspicuous consumption" (Veblen, T. B.). 5.) Conspicuous consumption has been analyzed as a primitive form of exchange by Mauss, M., and has been mentioned repeatedly in the intellectual lineage of Bataille, G., Baudrillard, J., et al. This action is concerned with the acquisition of social status in order to save one's own face, and shows what forms the depth-psychological base stratum in the "class system" of human beings, as well as illuminating the "social functions of dignity and class distribution." We have discovered a continuous course from the psychogenic experiences occurring at the onset and previously considered trifling and therefore ignored, proceeding to the symptoms and to the structure of the symptoms. In particular, by using a phenomenological analysis of shame and symptom structure, we show a new hypothesis about the structure of symptoms which is completely different from those created according to various previous opinions. Such understanding throws new light on this disease. PMID- 10810895 TI - [Reconsideration on "clinico-pathological study on the autopsy cases showing glove-and-stocking type sensory disturbance in relation to Minamata disease"]. PMID- 10810896 TI - [Progress report on changing the term "seisin bunretsu byou". Committee on concepts and terminology of psychiatric disorders sub-committee on the term schizophrenia]. PMID- 10810897 TI - [Neuropsychology of frontal type dementia]. AB - The clinical conceptual change in frontal type dementia is reviewed in discussing its relationships to several related concepts such as Pick's disease, frontotemporal dementia (FTD), semantic dementia (SD) and frontotemporal lobar degeneration. We analyzed frontal type dementia selected from a consecutive series of our outpatients as to the details of neuropsychological symptoms, psychiatric symptoms, and abnormal behaviors. In our series of 143 patients with primary degenerative dementia, there were 16 cases of FTD and 6 cases of SD. Patients with two types of FTD and patients with SD were not distinguishable by neuropsychological examinations, behavioral abnormalities and psychiatric symptoms assessed with the Neuropsychiatric Inventory except for aphasia. The clinical picture of frontal type dementia involves frontal lobe symptoms such as disinhibition, apathy and stereotypy. Semantic memory loss for words, objects or faces suggestive of temporal lobe involvement developed only in patients with SD, and not in patients with FTD. Certain behavioral symptoms seen in frontal type dementia may respond to selective serotonin reuptake inhibitors. In care for patients with frontal type dementia, behavioral disturbances can be diminished and the quality of life can be improved by using their preserved procedural memory, pathological stereotypic behavior and stimulus-bound behavior such as utilization behavior and environmental dependency syndrome. PMID- 10810898 TI - [Community support of psychiatric patients for their normalization]. PMID- 10810899 TI - [Role of psychiatric clinics--through a stage production activity]. PMID- 10810900 TI - [Regional cooperation in psychiatric care in future--a viewpoint of a university psychiatric department]. PMID- 10810901 TI - [Structural incompleteness and psychiatric care at a general hospital in a community]. PMID- 10810902 TI - [Conditions for regional cooperation --information sharing, freedom of selection, and equal cooperation]. PMID- 10810903 TI - [Organization of the second tier health services in future psychiatric care--a viewpoint of a psychiatric clinic]. PMID- 10810904 TI - [On the question of service areas in psychiatric care and welfare]. PMID- 10810905 TI - [Functions of general hospital psychiatric departments in the creation of the second tier in mental health services]. PMID- 10810906 TI - [Enhancement of resources for regional mental health promotion--in relation to the second tier in psychiatry and regional characteristics]. PMID- 10810907 TI - [Maintenance of local life activities and second tier area-- a viewpoint of the psychiatric health and welfare center]. PMID- 10810908 TI - [Organization of the second tier structure in mental health services]. PMID- 10810909 TI - [The hardness of the clinical treatment methods]. PMID- 10810911 TI - Perspectives of psychiatry in the 21st century. PMID- 10810910 TI - [Committee on Psychiatric Services and Law. The Japan Society of Psychiatry and Neurology]. PMID- 10810912 TI - [Psychopathological concepts of endogenous causes for mental diseases]. PMID- 10810913 TI - [Psychogenetics views on endogenous causes of mental diseases]. PMID- 10810914 TI - [Psychophysiological views on endogenous causes of mental diseases]. PMID- 10810915 TI - [Neurobiological views on endogenous causes of mental diseases: Influence of environment around fetal and neonatal stage on brain morphogenesis- predisposition for depressive disorders]. PMID- 10810916 TI - [Concept on endogenous causes of mental diseases from the point of view of genetically regulated environment]. PMID- 10810917 TI - Fighting the stigma of schizophrenia. PMID- 10810918 TI - [Psychiatry in Japan and the world--Morita Therapy and the other psychotherapies in the world]. PMID- 10810919 TI - [International cooperation of psychiatry and health care]. PMID- 10810920 TI - Wake-up call; time to take off. PMID- 10810921 TI - Peripheral interactions on lipid membrane surfaces. PMID- 10810922 TI - The electrostatics of lipid surfaces. AB - Charged lipids constitute a substantial fraction of all membrane lipids. Their charges vary in quantity and distribution within their headgroup regions. In long range interactions, their charges' value and electrostatic potential in the vicinity of the membrane surface can be approximated by the Guy-Chapman theory. This theory treats the interface as a charged structureless plain surrounded by uniform environments. However, if one considers intermolecular interactions, such assumptions need to be revised. The interface is in reality a thick region containing the residual charges of lipid headgroups. Their arrangement depends on the type of lipid present in the membrane. The variety of lipids and their biological functions suggests that charge distribution determines the extent and type of interaction with surface associated molecules. Numerous examples show that protein behavior at the lipid bilayer surface is determined by the type of lipid present, indicating protein specificity towards certain surface locations and local properties (determined by lipid composition) of a particular type. Such specificity is achieved by a combination of electrostatic, hydrophobic and enthropic effects. Comparing lipid biological activity, it can be stated that residual charge distribution is one of the factors of intermolecular recognition leading to the specific interaction of lipid molecules and selected proteins in various processes, particularly those involved with signal transduction pathways. Such specificity enables a variety of processes occurring simultaneously on the same membrane surface to function without cross-reaction interference. PMID- 10810923 TI - Significance of reduced dimensionality in reaction kinetics: impact of multi-site particles. AB - This review examines novel kinetic properties of enzymes on membrane surfaces or states of restricted diffusion. A leading feature is the presence of multiple enzymes and/or substrates per particle. In these states, enzymes can be influenced by parameters such as the number of substrates or enzymes per particle, particle size, the rates of exchange of substrate or enzyme from the particle, or substrate diffusion to the particle. These steps are independent of the enzyme site parameters which are described by classical enzymology. The results make it clear that non-classical behaviors are important to biological systems, are the basis for some enzyme expression levels and are determinants of cellular design. To identify more unique functions of these states, descriptions of catalysis in the non-solution state should become a part of kinetic education in biology. PMID- 10810924 TI - The influence of membrane lateral pressures on simple geometric models of protein conformational equilibria. AB - The function of many intrinsic membrane proteins requires a conformational transition that is often strongly influenced by the molecular composition of the bilayer in which the protein is embedded. Recently, a mechanism for this shift in conformational equilibrium was suggested, in which it is argued that a shift in distribution of lateral pressures of the bilayer resulting from a change in lipid composition alters the amount of mechanical work of the protein conformational transition, if the change in the cross-sectional area profile of the protein varies with depth within the bilayer. As there is little information on the change in shape of the transmembrane region of any protein, various simple geometric models are considered. For both a generic model, and more specific models that approximate likely cooperative rearrangements of alpha-helices in bundles, it is found that the conformational equilibrium depends on the first and second integral moments of the lateral pressure distribution. In addition to revealing the possible physical underpinnings of the well-known correlation between protein activity and the 'nonlamellar' tendency of bilayer lipids, this dependence on moments of the pressure profile allows for prediction of the relative effects of different lipid compositional changes even in the absence of information on specific protein shape changes. Effects of variation in acyl chain length, degree and position of cis-unsaturation, and addition of cholesterol and small interfacially-active solutes (n-alkanols) are compared. PMID- 10810925 TI - Fluorescent probes used to monitor membrane interfacial polarity. AB - The polarity of the interface between a lipid bilayer membrane and bulk water is an important physical parameter of the membrane. It is likely that several membrane-dependent biological functions are modulated by this property. However, interfacial polarity can be difficult to define because of an imprecise knowledge of the molecular nature of the interface. Nevertheless, attempts have been made to measure this quantity with the use of fluorescent probes which are sensitive to the solvent polarity. Often, however, other factors, such as the rate of solvent relaxation must be known in order to interpret the fluorescent properties in terms of the dielectric constant. In addition, the spatial orientation and location of the fluorophore are often not known precisely. Nevertheless, there have been successful efforts to gain a more accurate knowledge of this aspect of membrane physical properties and its relationship to biological phenomena is discussed. PMID- 10810926 TI - Hydration of lipid membranes and the action mechanisms of anesthetics and alcohols. PMID- 10810927 TI - Membrane phospholipid dynamics during cytokinesis: regulation of actin filament assembly by redistribution of membrane surface phospholipid. AB - To study molecular motion and function of membrane phospholipids, we have developed various probes which bind specifically to certain phospholipids. Using a novel peptide probe, RoO9-0198, which binds specifically to phosphatidylethanolamine (PE) in biological membranes, we have analyzed the cell surface movement of PE in dividing CHO cells. We found that PE was exposed on the cell surface specifically at the cleavage furrow during the late telophase of cytokinesis. PE was exposed on the cell surface only during the late telophase and no alteration in the distribution of the plasma membranebound peptide was observed during the cytokinesis, suggesting that the surface exposure of PE reflects the enhanced transbilayer movement of PE at the cleavage furrow. Furthermore, cell surface immobilization of PE induced by adding of the cyclic peptide coupled with streptavidin to prometaphase cells effectively blocked the cytokinesis at late telophase. The peptide-streptavidin complex bound specifically to cleavage furrow and inhibited both actin filament disassembly at cleavage furrow and subsequent plasma membrane fusion. Binding of the peptide complex to interphase cells also induced immediate disassembly of stress fibers followed by assembly of cortical actin filaments to the local area of plasma membrane where the peptide complex bound. The cytoskeletal reorganizations caused by the peptide complex were fully reversible; removal of the surface-bound peptide complex by incubating with PE-containing liposome caused gradual disassembly of the cortical actin filaments and subsequent formation of stress fibers. These observations suggest that the redistribution of plasma membrane phospholipids act as a regulator of actin cytoskeleton organization and may play a crucial role in mediating a coordinate movement between plasma membrane and actin cytoskeleton to achieve successful cell division. PMID- 10810928 TI - Controlling cytoskeleton structure by phosphoinositide-protein interactions: phosphoinositide binding protein domains and effects of lipid packing. AB - Cell movement and resistance to mechanical forces are largely governed by the cytoskeleton, a three-dimensional network of protein filaments that form viscoelastic networks within the cytoplasm. The cytoskeleton underlying the plasma membrane of most cells is rich in actin filaments whose assembly and disassembly are regulated by actin binding proteins that are stimulated or inhibited by signals received and transmitted at the membrane/cytoplasm interface. Inositol phospholipids, or phosphoinositides, are potent regulators of many actin binding proteins, and changes in the phosphorylation of specific phosphoinositide species or in their spatial localization are associated with cytoskeletal remodeling in vitro. This review will focus on recent studies directed at defining the structural features of phosphoinositide binding sites in actin binding proteins and on the influence of the physical state of phosphoinositides on their ability to interact with their target proteins. PMID- 10810929 TI - Biophysics of ceramide signaling: interaction with proteins and phase transition of membranes. AB - Ceramides have been implied in intracellular signal transduction systems regulating cellular differentiation, activation, survival and apoptosis and thus appear capable of changing the life style of virtually any cell type. Ceramide belongs to the group of sphingosine-based lipid second messenger molecules that are critically involved in the regulation of diverse cellular responses to exogenous stimuli. The emerging picture suggests that coupling of ceramide to specific signaling cascades is both stimulus and cell-type specific and depends on the subcellular topology of its production. However, little is understood about the molecular mode of ceramide action. In particular, in lieu of a defined ceramide binding motif it is not clear how ceramide would directly interact with putative target signaling proteins. This article proposes two modes of ceramide action. First, a protruding alkyl chain of ceramide may interact with a hydrophobic cavity of a signaling protein providing a lipid anchor to attach proteins to membranes. Second, the generation of ceramide generally increases the volume of hydrocarbon chains within the lipid bilayer thereby enhancing its propensity of to form a hexagonal II phase (Hex II). Besides the generation of a hydrophobic interaction site for proteins local hexagonal phase II formation can also change the membrane fluidity and permeability, which may impinge on membrane fusion processes, solubilization of detergent-resistant signaling rafts, or membrane receptor internalization. Thus, ceramide production by sphingomyelinases (SMase) can play a pivotal signaling role through direct interaction with signaling proteins or through facilitating the formation and trafficking of signal transduction complexes. PMID- 10810930 TI - Interactions of DNA with giant liposomes. AB - DNA interactions with the bilayers of cationic liposomes were studied using a novel model experiment: DNAs were locally injected by a micropipette to a part of a giant unilamellar vesicle. The resulting phenomena were directly observed in optical microscope. Giant unilamellar vesicles (GUVs), about 100 microm in diameter, made of phosphatidylcholines and up to 33 mol% of the natural bioactive cationic amphiphile sphingosine, were obtained by electroformation. The effects of DNAs of different length were tested: (i) 'short' DNAs-oligonucleotide 21b, and calf thymus 250 bp; (ii) 'long' DNAs-plasmid DNAs in super coil or liner form (between 2.7 and 8.0 kbp). DNAs were injected native, as well as marked with the fluorescent dye Hoechst. The resulting membrane topology transformations were monitored in phase contrast, while the DNA distribution was followed in fluorescence. DNA-induced endocytosis was observed due to the DNA/lipid membrane local interactions for all DNAs tested. Some of the DNA in the formed complex was associated with the induced endosomes, and some of it remained spread over the 'mother' GUV membrane for all DNAs tested, except for the longest one--the linear plasmid of 8 kbp. The last remained at the 'mother' GUV membrane and was not transported with the induced endosomes to the internal GUV space. Possible mechanisms for DNA/lipid membrane interaction were suggested. One of them involves DNA encapsulation within an inverted micelle included in the lipid membrane. The model observations could help in understanding events associated with interaction of DNA with biological membranes, as well as cationic liposomes/DNA complexes formation in gene transfer processes. PMID- 10810931 TI - Docetaxel (Taxotere) in the treatment of cancer. PMID- 10810932 TI - Docetaxel (Taxotere) as a single agent and in combination chemotherapy for the treatment of patients with advanced non-small cell lung cancer. AB - Docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) has high and reproducible single-agent activity in non-small cell lung cancer, inducing responses in nearly one third of patients treated first-line. As a second-line treatment, docetaxel is the only agent so far shown in randomized trials to prolong survival when compared with either vinorelbine or ifosfamide or best supportive care. When docetaxel is given on a weekly rather than once every 3 week schedule, the risk of neutropenia is reduced while activity appears to be maintained. The weekly schedule may prove to be a more favorable one for combination chemotherapy regimens. To date, docetaxel has been studied with other agents frequently using a once every 3 weeks schedule. Combinations of docetaxel with cisplatin, carboplatin, gemcitabine, or vinorelbine are generally well tolerated. With all four combinations, the pooled response rates in phase II studies are approximately 40%. A phase II study of 51 patients treated with the combination of 100 mg/m2 docetaxel and 900 mg/m2 gemcitabine reported a 13-month median survival. In a randomized phase II trial comparing docetaxel/gemcitabine with docetaxel/cisplatin, the two regimens had comparable efficacy and toxicity. Response rates of up to 50% have been reported using docetaxel in conjunction with vinorelbine. PMID- 10810933 TI - Docetaxel (Taxotere) in combination with anthracyclines in the treatment of breast cancer. AB - Considering the single-agent activity of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) and doxorubicin in breast cancer and their potential non cross-resistance, several docetaxel/anthracycline-based combination chemotherapies were developed in phase I and II programs for metastatic breast cancer patients. The rationale for these combinations was also reinforced by the fact that docetaxel showed significant activity in phase III trials in patients previously exposed or having failed anthracycline chemotherapy. In a pivotal randomized phase III study of doxorubicin plus docetaxel versus doxorubicin plus cyclophosphamide as first-line chemotherapy for 429 patients with metastatic breast cancer, doxorubicin/docetaxel emerged as the more effective regimen. Despite a lower-dose intensity of doxorubicin, patients receiving doxorubicin/docetaxel experienced a higher response rate as well as a significantly longer time to progression and time to treatment failure. This difference was seen even in patients with poor-prognosis disease. Febrile neutropenia was more common in doxorubicin/docetaxel-treated patients. However, there were no septic deaths among 213 patients receiving doxorubicin/ docetaxel. Extrahematologic toxicity appeared mild for both regimens and the combination docetaxel/doxorubicin did not increase the cardiac toxicity expected for an anthracycline-containing regimen. Docetaxel plus doxorubicin is the first regimen, involving a newly developed agent, proven superior to a standard anthracycline-containing combination in metastatic breast cancer and its potential is now being investigated in the adjuvant setting. PMID- 10810934 TI - Docetaxel (Taxotere) in HER-2-positive patients and in combination with trastuzumab (Herceptin). AB - In addition to being an important indicator of poor prognosis, human epidermal growth factor receptor-2 (HER-2) status may help identify those patients in whom chemotherapy is the most appropriate choice of therapy. In several studies, including a trial of sequential neoadjuvant therapy in which certain patients received docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) following four courses of cyclophophamide 1000 mg/m2, doxorubicin 50 mg/m2, vincristine 1.5 mg/m2 on day one, and prednisone 40 mg by mouth for 5 days, HER-2 positivity predicted response (including pathologic response) to chemotherapy. In vitro and in vivo, docetaxel has demonstrated true synergy with the recombinant human anti HER-2 monoclonal antibody trastuzumab (Herceptin; Genentech, San Francisco, CA). In a phase III study comparing trastuzumab alone with trastuzumab plus chemotherapy (either paclitaxel or doxorubicin plus cyclophosphamide), combining the antibody with cytotoxic drugs increased response duration, time to progression, and survival in first-line metastatic breast cancer patients. Preliminary clinical data suggest that the combination of trastuzumab with docetaxel is active and well tolerated, and pilot studies of adjuvant therapy using trastuzumab and docetaxel combinations are under way in high-risk patients. PMID- 10810936 TI - Contact lens research in 1999. PMID- 10810935 TI - Docetaxel (Taxotere) in hormone-refractory prostate cancer. AB - Considerations of both molecular biology and data from in vitro studies suggest a potential for the combination of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) with estramustine in the treatment of patients with hormone refractory prostate cancer. Based on data from two phase I studies, the docetaxel dose recommended for phase II study in combination with estramustine in minimally and extensively pretreated patients is 70 mg/m2 and 60 mg/m2, respectively. The dose-limiting toxicity was neutropenia. In one phase I study, 63% of the 34 patients treated showed at least a 50% decline in prostate-specific antigen. An objective response was seen in 28% of patients with measurable disease, and overall median survival (22.8 months) is highly encouraging. In the second study, 82% of 17 patients showed a greater than 50% decline in prostate-specific antigen and, at the 70 mg/m2 dose level, two of six patients showed prostate-specific antigen normalization. Phase II studies have demonstrated more than 50% declines in 59% to 88% of patients treated at 70 mg/m2. Although reduction of the dose of estramustine appears to result in a somewhat lower response rate, the contribution made by estramustine to the efficacy of the estramustine/docetaxel combination must be established by randomized trials. Dexamethasone, however, does not appear to significantly contribute to the response rate of estramustine and docetaxel. Phase III studies comparing docetaxel plus estramustine with mitoxantrone plus corticosteroids are currently being planned. If the promise of docetaxel hormone-refractory prostate cancer is realized, it may be appropriate to design clinical trials that evaluate docetaxel-based regimens as adjuvant therapy in patients who are at a high risk for relapse after definitive local therapy. PMID- 10810937 TI - Methods used to evaluate the effectiveness of contact lens care solutions and other compounds against Acanthamoeba: a review of the literature. AB - PURPOSE: The purpose of this paper is to review the literature concerning the methods used to evaluate contact lens care solutions against Acanthamoeba. Acanthamoeba keratitis is a potential threat, with 85% of the cases being reported in contact lens wearers. METHODS: Several studies from the published literature that evaluated contact lens disinfectants were reviewed. The variables included test organism, strain and morphology, growth conditions, inoculum preparation, inoculation method, test solutions and concentration, contact time, neutralization, recovery, quantitation method, and viability determination of survivors. The methods used to test Acanthamoeba against the disinfectants were compared and contrasted. RESULTS: After a thorough review of methods used to test Acanthamoeba, it was found that there is great variability in the methods used to evaluate contact lens disinfectants. The majority of the studies used A.castellanii and A.polyphaga cysts grown axenically in PYG medium containing cations at about 30 degrees C and the inoculum contained about 1.0 x 10(5) cells/mL. Inactivation media or centrifugation of cells was used to neutralize test samples. Quantitation was performed in most studies and viability was checked in all studies. The disinfectants tested most often were PHMB, hydrogen peroxide, thimerosal, and chlorhexidine. CONCLUSIONS: After reviewing the studies presented here it can be concluded that an effective method for testing Acanthamoeba against contact lens disinfectants would include A.castellanii or A.polyphaga grown axenically in PYG containing cations and a concentration of organisms high enough to adequately measure kill, a neutralization step, recovery and quantitation of organisms followed by a viability check of survivors. PMID- 10810938 TI - Hydrogel contact lens ageing. AB - PURPOSE: To determine if hydrogel lens water content decreases over wearing periods of up to four weeks; that is, demonstrate an 'ageing' effect. METHODS: Six subjects (4 males, 2 females; age range: 34.8 +/- 11.0 years) wore each of the following lenses for one month: Acuvue, Frequency 38, Gentle Touch and Proclear. Pre-insertion and post-removal lens water content was measured daily by the subjects using an Atago CL-1 Soft Contact Lens Refractometer for a 28 day period. Post-insertion and pre-removal comfort was recorded over the same period. RESULTS: A significant lens ageing effect was noted whereby the pre-insertion lens water content decreased significantly over the 28 day cycle for all lenses (P < 0.05). The pattern for daily dehydration remained consistent for the 28 day period for the Acuvue lens and the Gentle Touch lens but decreased for the Frequency 38 lens and the Proclear lens. Comfort decreased significantly by the end of the day for three of the four lens types investigated. CONCLUSIONS: It is apparent that the extent of lens dehydration changes over time, and that this pattern of change is different for different lens types. The associated loss of oxygen performance with dehydration is demonstrated. These data will assist practitioners in understanding the clinical effects of soft lens dehydration over time. PMID- 10810939 TI - Efficiency and accuracy of two computerized topography software systems for fitting rigid gas permeable contact lenses. AB - PURPOSE: The world of computerized videokeratoscopy is continually evolving. Computerized videokeratoscope contact lens programs boast the capability to accurately design final rigid gas permeable (RGP) lens parameters and simulate contact lens fluorescein patterns. We evaluated the EyeSys System 2000 Pro-Fit software (v. 4.0) and Humphrey Atlas MasterVue Contact Lens Module (v. A6) for efficiency and accuracy in fitting RGP contact lenses on normal subjects. METHODS: Balanced manifest refractions, slit lamp examinations, and computerized topographical analysis with RGP module lens fitting were performed and compared for all eyes. Lens parameters were chosen after an optimal simulated fluorescein pattern was achieved by either accepting the initially recommended default lens or a modification. Final lens parameters were based on clinical performance. RESULTS: With the Humphrey Atlas software, which uses a default topographic tear clearance fitting protocol set by the manufacturer, 24/50 (48%) eyes required modifications to the default fitting parameters based on the simulated fluorescein patterns. Nine of 50 eyes (18%) required further modifications once the RGP lens was dispensed. The EyeSys Pro-Fit software, which allows a custom tear clearance fitting protocol set by the user as default, required modifications to 20/35 eyes (57.1%) from default fittings provided by the system. Six of 35 eyes (17.1%) required subsequent modifications once the RGP lens was dispensed. Comparison of the EyeSys to Humphrey modules revealed no significant difference in efficiency (P=0.51), defined as any changes required to the default parameters; however, the user made more modifications to the EyeSys module from the default setting. There was no significant difference in final success rates between EyeSys (82.9%) and Humphrey (82%) systems (P=1.00). CONCLUSION: Although both topography guided contact lens modules offered accuracy in RGP fitting, a user programmed default (EyeSys) protocol did not improve efficiency in RGP lens parameter selection compared to a manufacturer programmed default module (Humphrey). The need for a knowledgeable contact lens fitter remains to guide the systems in achieving the optimal fit. PMID- 10810940 TI - Is intraoperative topography predictive of postoperative topographical changes following refractive surgery? AB - PURPOSE: To evaluate the usefulness of measuring corneal topography intraoperatively by Intraoperative PAR Corneal Topography System (IOPAR CTS) to predict the postoperative topographical changes in patients undergoing refractive surgery. METHODS: Topographical measurements of 9 eyes of 9 patients were taken by IOPAR CTS: 6 immediately after Intrastromal Corneal Ring Segments (ICRS) placement surgery and 3 immediately after Photorefractive Keratectomy (PRK) surgery. The topographical images taken by IOPAR CTS were analyzed to determine the quality of the image produced and the correlation of the keratometric data from the central 3 mm of cornea with measurements taken from an autokeratometer (Auto-Km) and the EyeSys Corneal Analysis System (CAS) for the same eyes taken three months following the procedure. RESULTS: At the 3-month postoperative exam, the correlation coefficients for flat (K1), steep (K2), and average keratometry (AK) readings produced by the Auto-Km and the EyeSys CAS were quite high. However, the correlation coefficients for K1, K2, AK, As or steep meridian axis (Ax) readings taken intraoperatively by IOPAR CTS were quite low when compared to the readings taken by either the Auto-Km or EyeSys CAS three months postoperatively. CONCLUSION: We found IOPAR CTS, relatively difficult to perform in the operating room during refractive surgical procedures and not useful clinically in predicting the postoperative topographical changes induced by ICRS or PRK procedure. Further studies are required to establish the efficacy and reproducibility of the instrument. PMID- 10810942 TI - Contact lens fitting difficulties following refractive surgery for high myopia. AB - PURPOSE: To describe the clinical and optical problems encountered in contact lens fitting following refractive surgery for high myopia. METHODS: Following refractive surgery for high myopia (greater than -10.00 D) we corrected residual refractive errors with contact lenses in the four eyes of two patients. The first patient had undergone bilateral laser in situ keratomileusis (LASIK),with two subsequent LASIK retreatments in the left eye. Ten months later she was fit with rigid gas permeable (RGP) lenses in both eyes. The second patient had undergone a clear lens extraction in the right eye and radial keratotomy followed by photorefractive keratectomy(PRK) in the left eye. She was fit with toric soft lenses six years postoperatively. RESULTS: Final visual acuity obtained with contact lenses was 20/25-20/20 in all eyes. The first patient required significant minus lens power compensation. Furthermore, the RGP lens in the left eye was slightly decentered due to corneal irregularity induced by LASIK. The second patient had regular corneal surfaces and was successfully fit with daily wear toric soft lenses despite the 2.75 D of residual astigmatism in the left eye. CONCLUSIONS: Following refractive surgery for high myopia a proportion of patients will remain undercorrected. In these patients the alterations in corneal architecture that ensue make contact lens fitting more challenging. Patients with regular astigmatism may be fitted successfully with toric soft lenses. Patients with corneal irregularities should be fit with RGP lenses. PMID- 10810941 TI - Characteristics of the myopic patient population applying for refractive surgery. AB - PURPOSE: Assessment of the characteristics of the myopic patient population applying for refractive surgery in order to determine the potential market for myopic refractive surgery. METHODS: Records of consecutive patients who responded to an advertisement for refractive surgery to correct myopia were evaluated retrospectively with regards to patient demographics and the amount and distribution of the refractive error. Data were compared to that available from population-based statistics for distribution of myopia in the general population. For statistical analysis, one sample Student's t-test and two tailed Student's t test were utilized. RESULTS: Two hundred fifty seven patients (140 women and 117 men) responded to an advertisement for refractive surgery during the six month period between January and June 1998. Mean spherical equivalent (SEQ) of the patient population was -4.59+/-2.54 D (min;-0.25 D, max;-15.75 D) right eye (OD) and -4.62+/-2.82 D (min;-0.25 D, max;-15.25 D) left eye (OS). Among the patients who had myopia with an astigmatism of at most 1.00 D (n=165), the distribution of refractive error was statistically significantly different from that obtained from population-based statistics, such that, although most of the myopic population (40%) had an SEQ of -1.00 to -2.25 D, the majority of our patients (54.8%) who applied for myopic refractive surgery had an SEQ of -2.50 to -5.00 D. Another striking difference was that, although patients with an SEQ more than 6.00 D were a minority(2%) in the population study, in our study group, they comprised 16.7% of the patients seeking refractive correction. The difference between the SEQ of the right and left eyes ranged from 0.00 D to 13.0 D (mean, 0.89+/-1.5 D), 47.1 % having a difference of at most +/-0.5 D between the two eyes. The mean cylindrical error in the patient population was 0.69+/-0.93 D (min: 0, max: -4.5) OD and 0.69+/-0.96 D (min: 0, max: -4.5) OS. There were no statistically significant differences between the distribution of SEQ or cylindrical refractive error between males and females. CONCLUSION: Although a population-based study reported that most of the myopic population (40%) had an SEQ of -1.00 to -2.25 D, the majority of our patients (54.8%) who applied for myopic refractive surgery had an SEQ of -2.50 to -5.00 D. On the other hand, while patients with an SEQ of -6.00 D and more constituted only about 2% of the general population, they accounted for 16.7% of our study population. Therefore, the refractive characteristics of the patient population applying for myopic refractive surgery may not necessarily parallel that of general population-based statistics. In order to establish a more effective refractive surgery practice, it is feasible to perform local studies and reevaluate the requirements of your practice accordingly. PMID- 10810943 TI - Ophthalmia nodosa caused by casual handling of a tarantula. AB - PURPOSE: To present the diagnostic and therapeutic challenges of ophthalmia nodosa secondary to tarantula hairs. METHODS: We present a case of a 28-year-old male with a one-week history of skin rash followed by ocular irritation, and blurring of vision after handling a Chilean rose tarantula. A Medline-guided literature search was performed to review the current and historical knowledge of ophthalmia nodosa. The clinical presentation and therapeutic options in ophthalmia nodosa were reviewed. RESULTS: Ophthalmia nodosa is a granulomatous, nodular reaction to vegetable or insect hairs including tarantula hairs. Tarantula hairs can cause inflammation in all levels of the eye, from conjunctiva to retina. Therapy includes surgical removal of offending hairs and medical management of inflammation. CONCLUSIONS: Although tarantulas are considered harmless pets, ophthalmia nodosa is a potential danger when handling a tarantula. Furthermore the management of exposure to tarantula hairs can be difficult. PMID- 10810944 TI - Racial differences in breast cancer mortality. PMID- 10810945 TI - Despite financial penalties, French physicians' knowledge of regulatory practice guidelines is poor. AB - OBJECTIVE: To evaluate the level of awareness and knowledge of regulatory practice guidelines (references medicales opposables [RMOs] or regulatory medical references) implemented to control ambulatory care costs among French family physicians. DESIGN: Observational study. Participants were asked to identify RMO topics among a list of actual and fictitious RMO topics and the RMOs themselves among a list of actual and fictitious RMOs. SETTING: General practice in France. SUBJECTS: Three hundred twenty-one family physicians. MAIN OUTCOME MEASURE: Average score of 100 (95% confidence interval [CI]) on the awareness of RMO topics and knowledge of the RMOs. RESULTS: The average overall score was 55.8 of 100 (95% CI, 53.3-58.3) for the awareness of the RMO topics and 50.5 (95% CI, 48.3-52.7) for knowledge of the RMOs themselves-53.2 (95% CI, 51.1-55.3) for diagnostic RMOs and 47.8 (95% CI, 45.6-50.0) for therapeutic RMOs. Chance would have yielded an expected mean score of 50. A statistically significant difference was noted between the average score for actual (62.2) and fictitious (43.2) RMOs, P<.001. None of the respondents correctly identified all 24 correct answers. CONCLUSION: Despite implementation of RMO policy, the awareness and knowledge of RMOs among French family physicians seem weak. The number of RMOs and the difficulties in controlling physicians probably explain these results. Thus, it is doubtful that the RMO policy will have a long-term effect on physicians' behavior. PMID- 10810946 TI - Flexible sigmoidoscopy training and its impact on colorectal cancer screening by primary care physicians. AB - BACKGROUND: Colorectal cancer is an ideal disease for prevention with screening programs. Efforts to increase compliance with screening recommendations have included training primary care physicians to perform flexible sigmoidoscopy. OBJECTIVE: To assess the impact of flexible sigmoidoscopy training on compliance with current screening recommendations. METHODS: We performed a cross-sectional study of 232 patients cared for by physicians in a primary care network. MAIN OUTCOME MEASURES: Rates of screening for colorectal cancer and rates of undergoing flexible sigmoidoscopy were compared across patient groups according to the physician's training and whether the physician performs flexible sigmoidoscopy in his or her practice. RESULTS: Among 217 patients included in the analysis, 122 (56%) were cared for by physicians who were trained in flexible sigmoidoscopy, of whom 79 (36%) were cared for by physicians who perform flexible sigmoidoscopy in their practice. Patients cared for by physicians trained in flexible sigmoidoscopy were not significantly more likely to receive any colorectal cancer screening than were patients cared for by physicians not trained in flexible sigmoidoscopy (odds ratio, 1.16; 95% confidence interval, 0.67-2.01). However, patients cared for by physicians who perform flexible sigmoidoscopy in their practice were more likely to have undergone any colorectal cancer screening (odds ratio, 1.73; 95% confidence interval, 1.02-2.95) and flexible sigmoidoscopy (odds ratio, 2.69; 95% confidence interval, 1.14-6.36). CONCLUSION: Performance of flexible sigmoidoscopy by primary care physicians has the potential to increase the rate of colorectal cancer screening with flexible sigmoidoscopy. PMID- 10810947 TI - How does physician advice influence patient behavior? Evidence for a priming effect. AB - OBJECTIVE: To explore a potential "priming effect" of physician advice on patient responses to behavioral change interventions. DESIGN: Randomized controlled trial with a 3-month follow-up. SETTING: Four community-based group family medicine clinics in southeastern Missouri. PARTICIPANTS: Adult patients (N = 915). INTERVENTIONS: Printed educational materials designed to encourage patients to quit smoking, eat less fat, and increase physical activity. MAIN OUTCOME MEASURES: Recall, rating, and use of the educational materials; changes in smoking behavior, dietary fat consumption, and physical activity. RESULTS: Patients who received physician advice to quit smoking, eat less fat, or get more exercise prior to receiving intervention materials on the same topic were more likely to remember the materials, show them to others, and perceive the materials as applying to them specifically. They were also more likely to report trying to quit smoking (odds ratio [OR] = 1.54, 95% confidence interval [CI] = 0.95-2.40), quitting for at least 24 hours (OR = 1.85, 95% CI = 1.02-3.34), and making some changes in diet (OR = 1.35, 95% CI = 1.00-1.84) and physical activity (OR = 1.51, 95% CI = 0.95-2.40). CONCLUSIONS: Findings support an integrated model of disease prevention in which physician advice is a catalyst for change and is supported by a coordinated system of information and activities that can provide the depth of detail and individualization necessary for sustained behavioral change. PMID- 10810948 TI - Deliveries by family physicians in Connecticut: results of a practice-based research network. AB - OBJECTIVE: To obtain descriptive data on deliveries performed by family physicians in Connecticut during 1 year. DESIGN: All family physicians in Connecticut who provided obstetric care were personally contacted to enlist their participation in a research network. All such physicians agreed to participate and were instructed to mail a 15-item delivery data card to the author following each delivery that they participated in during calendar year 1996. MAIN OUTCOME MEASURES: Method of delivery, use of obstetric intervention (eg, oxytocin administration, episiotomy, and epidural anesthesia), and obstetric consultation rates. RESULTS: Thirty-two physicians, representing 9.0% of the family practice workforce in Connecticut, provided obstetric care. These 32 physicians, who practice in only 7 of the 31 acute care hospitals in the state, delivered 478 neonates during 1996. Most deliveries were by the spontaneous vaginal route, with forceps and vacuum used in 2.1% and 5.0% of vaginal deliveries, respectively. The primary cesarean section rate of these family physicians was low at 5.6% compared with a statewide rate for all providers (physicians and certified nurse midwives) of 12.4% (odds ratio, 0.42; 95% confidence interval, 0.28-0.63). Rates of certain obstetric interventions, such as oxytocin administration, epidural anesthesia, and episiotomy, varied greatly by hospital. Obstetric consultation rates also varied greatly among hospitals, ranging from 12.8% to 49.6%. CONCLUSIONS: Family physician involvement in maternity care in Connecticut is low. This study confirms a low rate of instrument-assisted and cesarean delivery in births attended by family physicians. Use of obstetric interventions and obstetric consultation varies greatly within the state. PMID- 10810949 TI - Clinical and demographic predictors of late-stage cervical cancer. AB - BACKGROUND: Despite increasingly widespread use of the Papanicolaou smear, almost half of all women with invasive cervical cancer are diagnosed at a late stage (regional or distant). Little is known about factors associated with late-stage diagnosis of cervical cancer. OBJECTIVE: To examine the relationship of age, race, education level, income level, smoking, marital status, health insurance type, comorbidity, and residence in an urban or rural setting to late stage at diagnosis of cervical cancer. METHODS: Incident cases of invasive cervical cancer occurring in 1994 in Florida were identified from the state tumor registry (N = 852). Cases were linked with state discharge abstracts and the 1990 US census. Multiple logistic regression was used to determine the relationship between predictor variables (age, race or ethnicity, marital status, smoking status, education level, income level, insurance type, comorbidity, and urban vs rural residence) and the odds of late-stage diagnosis. RESULTS: Age, marital status, and insurance type were associated with late-stage diagnosis. Each additional year of age was associated with a 3% increased odds of late-stage diagnosis (odds ratio [OR], 1.03; 95% confidence interval [CI], 1.02-1.05; P<.001). Being unmarried was associated with a 63% increased odds of late-stage diagnosis (OR, 1.63; 95% CI, 1.18-2.25; P=.003). Being uninsured was associated with a 60% increased odds of late-stage diagnosis (OR, 1.60; 95% CI, 1.07-2.38; P=.02). Having commercial health maintenance organization insurance was associated with a 46% decreased odds of late-stage disease (OR, 0.54; 95% CI, 0.30-0.96; P=.04). Race, education level, income level, smoking status, comorbidity, and urban residence were not associated with stage at diagnosis. CONCLUSIONS: Women with cervical cancer who are elderly, unmarried, and uninsured are more likely to be diagnosed at a late stage. These women should be targeted for cervical cancer education and screening programs. PMID- 10810950 TI - Fragmentation of patient care between chiropractors and family physicians. AB - BACKGROUND: Most patients using alternative practitioners also receive care from physicians. It is unclear, however, how well alternative practitioners and physicians communicate and coordinate the care of shared patients. OBJECTIVE: To describe the communication and coordination of care for shared patients between chiropractors and family physicians as well as potential barriers to effectively sharing care. DESIGN, SETTING, AND PARTICIPANTS: A cross-sectional national random sample survey of 400 chiropractors and 400 family physicians. MAIN OUTCOME MEASURES: Reports on shared patients including information on adverse events, treatment, and health status. Attitudes toward perceived expertise as well as perceived liability and economic competition involved in sharing care were also assessed. RESULTS: Surveys were completed by 360 (49%) of the 736 eligible practitioners, including 227 chiropractors and 133 family physicians. Although a high degree of interaction occurs between the practitioners, family physicians received information from chiropractors on 26.5% of referred patients while chiropractors received information from family physicians in 25.0% of cases (P = .73). Both groups believed that they did not receive enough information on adverse health outcomes or treatment plans for shared patients. Although neither group was particularly oriented toward wanting to share care, family physicians were much less likely than chiropractors to feel comfortable sharing care (P<.001). CONCLUSIONS: These findings indicate that care is fragmented between chiropractors and the general medical sector, with little information communicated between health care providers on issues with critical importance to quality of care. Further study is needed to identify ways to improve communication and coordination of care. PMID- 10810951 TI - Physical health consequences of physical and psychological intimate partner violence. AB - BACKGROUND: Past studies that have addressed the health effects of intimate partner violence (IPV) have defined IPV as violence based on physical blows that frequently cause injuries. To our knowledge, no epidemiologic research has assessed the physical health consequences of psychological forms of IPV. OBJECTIVE: To estimate IPV prevalence by type and associated physical health consequences among women seeking primary health care. DESIGN: Cross-sectional survey. SETTING AND PARTICIPANTS: A total of 1152 women, aged 18 to 65 years, recruited from family practice clinics from February 1997 through January 1999 and screened for IPV during a brief in-clinic interview; health history and current status were assessed in a follow-up interview. RESULTS: Of 1152 women surveyed, 53.6% ever experienced any type of partner violence; 13.6% experienced psychological IPV without physical IPV. Women experiencing psychological IPV were significantly more likely to report poor physical and mental health (adjusted relative risk [RR], 1.69 for physical health and 1.74 for mental health). Psychological IPV was associated with a number of adverse health outcomes, including a disability preventing work (adjusted RR, 1.49), arthritis (adjusted RR, 1.67), chronic pain (adjusted RR, 1.91), migraine (adjusted RR, 1.54) and other frequent headaches (adjusted RR, 1.41), stammering (adjusted RR, 2.31), sexually transmitted infections (adjusted RR, 1.82), chronic pelvic pain (adjusted RR, 1.62), stomach ulcers (adjusted RR, 1.72), spastic colon (adjusted RR, 3.62), and frequent indigestion, diarrhea, or constipation (adjusted RR, 1.30). Psychological IPV was as strongly associated with the majority of adverse health outcomes as was physical IPV. CONCLUSIONS: Psychological IPV has significant physical health consequences. To reduce the range of health consequences associated with IPV, clinicians should screen for psychological forms of IPV as well as physical and sexual IPV. PMID- 10810952 TI - Primary care physician incentives in medical group practices. AB - CONTEXT: Although medical groups are adapting to changes in financing health care, little is known about individual physician incentives in this environment. OBJECTIVES: To describe methods group practices use to compensate primary care physicians in a managed care environment and to examine the association of revenue sources for the group practice from all patients and primary care physician incentives. DESIGN: We surveyed by mail group practice administrators for practices that had at least 200 members continuously enrolled in 1995. SETTING: Group practices that had contractual arrangements with Blue Cross/Blue Shield of Minnesota. PARTICIPANTS: One hundred of 129 group practices returned usable surveys. RESULTS: Most groups had some portion of primary care physicians' compensation at risk, although 17 groups compensated them through fully guaranteed annual salary. Seventy-one groups used productivity, 4 groups used quality of care, 1 group used utilization, and 30 used group financial performance. Factors reported to significantly influence primary care physician compensation included billings or charges, overall group practice performance, and net revenue or profit. Groups that had a higher proportion of income from various types of fee-for-service arrangements used lower proportions of base salary for primary care physician compensation and were more likely to relate physician income to measures of productivity. CONCLUSIONS: Substantial variation exists in the types of primary care physician incentives implemented by medical groups. Base salary, individual productivity, and group financial performance were most frequently used to determine compensation. Physician personal financial risk was higher overall in group practices that derived more revenue from fee-for service contracts. PMID- 10810953 TI - The adult with cerebral palsy. AB - Important aspects of eliciting a proper history, obtaining a review of systems, and performing a physical examination in adults with cerebral palsy are presented. Information regarding diagnosis, etiology, and epidemiology of cerebral palsy as well as suggestions for performing examinations and procedures on uncooperative and extremely dysmorphic patients are included. A MEDLINE search of all English-language publications related to cerebral palsy from 1985 to 1999 was conducted. Other older references also were obtained from articles published during this period. Our personal experiences in caring for a group of approximately 300 adults with cerebral palsy and other developmental disabilities in specialized centers for nearly a decade are used frequently throughout this review. Emphasis is given to studies of adults. Studies of children are included because there is a lack of data on adults. These studies are identified as such in the text, with extrapolation to adults only where there is a sound clinical or scientific basis. The number of adults with cerebral palsy is increasing. This growth is due to increased survival of low-birth-weight infants and increased longevity of the adult population. Depending on clinical status and the age at which survival is calculated, 65% to 90% of children survive until adulthood. Despite these observations, there is a lack of information in the literature and a lack of relevant post-graduate training programs for physicians in the adult health care system. PMID- 10810954 TI - Treating asymptomatic bodily contacts of patients with scabies. PMID- 10810955 TI - Following depression in primary care: do family practice physicians ask about depression at different rates than internal medicine physicians? AB - OBJECTIVE: To determine whether the chronically or recurrently depressed patients of family practice and internal medicine physicians differed in the proportion reporting that their primary care physician asked them about depression symptoms. DESIGN: A cross-sectional observational study of chronically or recurrently depressed survey respondents who identified a family practice or internal medicine physician as their primary care provider. SETTING: A large not-for profit group-model health maintenance organization in the northwestern United States, with a population representative of its service area. PATIENTS: Health maintenance organization members (n= 1161) with ongoing or recurring depression or dysthymia who responded to a 1993 survey and who identified either a family practice or internal medicine physician as their primary care provider. MAIN OUTCOME MEASURE: Patients' self-report of their primary care physician asking them: (1) whether they had been feeling sad, blue, or depressed; (2) to fill out a questionnaire about their mood or feelings; and (3) whether they had been thinking about death or suicide. RESULTS: Chronically or recurrently depressed patients of family practice physicians were more likely to report that their physician asked them about depressive symptoms than were patients of internal medicine physicians (34.0% vs 27.3%) (P=.02). This finding persisted in a multivariate analysis. CONCLUSION: Family practice physicians may be more attentive to depressive disorders than internal medicine physicians. PMID- 10810956 TI - Neuropeptides, neurotransmitters, body weight, and menopause. PMID- 10810957 TI - Urogenital atrophy and low-dose vaginal estrogen therapy. PMID- 10810958 TI - The North American Menopause Society 1998 menopause survey: Part II. Counseling about hormone replacement therapy: association with socioeconomic status and access to medical care. AB - OBJECTIVE: The purpose of this study was to examine two predictors of women's obtaining hormone replacement therapy (HRT) counseling: socioeconomic status and access to health care. DESIGN: During May-July 1998, by means of random-digit telephone dialing, 749 postmenopausal women who were living in the United States and aged 50-65 years were interviewed. On average, they were 56.8 years and 11.8 years postmenopausal. Most (86.0%) were Caucasian, and their median annual income was approximately $40,000. Nearly all (90.8%) had medical insurance coverage; 47.6% of those insured received care from a managed care organization. Access to medical care was evidenced by 92.3% being under the care of a primary care physician, 92.3% ever having had a mammogram, 96.9% ever having had a pelvic examination, and 91.1% ever having had a serum cholesterol determination. RESULTS: Of these 749 women, 75.4% reported that they had received counseling about post-menopausal HRT from healthcare providers. Both level of education and level of income were associated with an increased likelihood that HRT counseling would be obtained. Having a personal physician, and particularly receiving care from a gynecologist, increased the likelihood that counseling would be available. There were no substantial differences in counseling frequency between women in managed care plans and those having other types of health insurance. In a multivariate model, adjusted odds ratios for receiving HRT counseling were 2.9 (95% confidence interval [CI] = 1.7-4.8) for having an annual income of $50,000 or more versus less than $30,000, 2.8 (95% CI = 1.7-4.5) for receiving care from a gynecologist versus other primary care physician, 1.9 (95% CI = 1.1-3.2) for being Caucasian versus not, and 1.5 (95% CI 1.0-2.2) for having a hysterectomy versus not. CONCLUSIONS: Three quarters of a sample of US postmenopausal women aged 50-65 years reported that they had been counseled about HRT. However, women of lowest socioeconomic status and those who did not have a primary care physician were least likely to have received counseling. No differences were observed in prevalence of counseling between women in managed care settings and those with other types of health insurance. The findings suggest that special efforts are necessary to provide menopause education and counseling to underserved women. PMID- 10810959 TI - Relationship among leptin, neuropeptide Y, and galanin in young women and in postmenopausal women. AB - OBJECTIVE: To determine whether hormonal status may affect neuropeptide Y (NPY), galanin, and leptin release in postmenopausal women and in young women. DESIGN: Forty-eight postmenopausal women aged 47-65 years and 35 young women aged 26-39 years were investigated. RESULTS: Plasma leptin concentrations increased with increasing body mass index in both young and postmenopausal women and were significantly higher in obese postmenopausal women than in obese young women (p < 0.01). Plasma NPY levels in obese young and postmenopausal women were significantly higher than in lean women (p < 0.01 and p < 0.01, respectively) and were significantly higher in obese and nonobese postmenopausal women than in young women (p < 0.05 and p < 0.001, respectively). Plasma galanin levels in postmenopausal women, both lean and overweight, were significantly lower than in young women (p < 0.01 andp < 0.01, respectively). In obese postmenopausal women, plasma galanin concentrations were lower without differing significantly from those in obese young women. However, they were significantly higher than that in lean postmenopausal women (p < 0.001). CONCLUSIONS: Our results suggest that the differences is plasma leptin, NPY, and galanin between postmenopausal women and young women may be related to body mass index rather than to differences in hormonal status and that the higher NPY levels in both lean and obese postmenopausal women than in young women indicate that factors other than body mass index may be involved. PMID- 10810960 TI - 17beta-estradiol vaginal tablet versus conjugated equine estrogen vaginal cream to relieve menopausal atrophic vaginitis. AB - OBJECTIVES: The efficacy and safety of 25-microg 17beta-estradiol vaginal tablets (Vagifem) were assessed and compared with 1.25-mg conjugated equine estrogen vaginal cream (Premarin Vaginal Cream) for the relief of menopausal-derived atrophic vaginitis, resulting from estrogen deficiency. DESIGN: In a multicenter, open-label, randomized, parallel-group study, 159 menopausal women were treated for 24 weeks with either vaginal tablets or vaginal cream. Efficacy was evaluated by relief of vaginal symptoms and concentrations of serum estradiol and follicle stimulating hormone. Safety was monitored by the incidence of adverse events, evaluation of endometrial biopsies, and clinical laboratory results. Patients also assessed the acceptability of the study medications. RESULTS: Composite scores of vaginal symptoms (dryness, soreness, and irritation) demonstrated that both treatments provided equivalent relief of the symptoms of atrophic vaginitis. At weeks 2, 12, and 24, increases in serum estradiol concentrations and suppression of follicle-stimulating hormone were observed in significantly more patients who were using the vaginal cream than in those who were using the vaginal tablets (p < 0.001). Fewer patients who were using the vaginal tablets experienced endometrial proliferation or hyperplasia compared with patients who were using the vaginal cream. Significantly more patients who were using the vaginal tablets rated their medication favorably than did patients who were using the vaginal cream (p < or = 0.001). Patients who were receiving the vaginal tablets also had a lower incidence of patient withdrawal (10% versus 32%). CONCLUSIONS: Treatment regimens with 25-microg 17beta-estradiol vaginal tablets and with 1.25-mg conjugated equine estrogen vaginal cream were equivalent in relieving symptoms of atrophic vaginitis. The vaginal tablets demonstrated a localized effect without appreciable systemic estradiol increases or estrogenic side effects. Vaginal tablet therapy resulted in greater patient acceptance and lower withdrawal rates compared with vaginal cream therapy. PMID- 10810961 TI - Estrogen replacement therapy and gallbladder disease in postmenopausal women. AB - OBJECTIVE: To examine the role of estrogen replacement therapy on the development of gallbladder disease in postmenopausal women. DESIGN: Systematic review of the English literature was conducted. All studies that addressed the association between hormone replacement therapy and gallbladder disease published from 1970 to the present were reviewed. RESULTS: Seven observational studies, two clinical trials, two case series, and one nonrandomized and three randomized investigations were reviewed. The results of each study were reported and analyzed. CONCLUSIONS: Estrogen replacement therapy in postmenopausal women increased the chances for gallstone formation. PMID- 10810962 TI - Higher levels of plasma estradiol and testosterone in healthy elderly men compared with age-matched women may protect aspects of explicit memory. AB - OBJECTIVE: To assess longitudinally the relationships between plasma levels of estradiol (E2) and free testosterone (T) and cognitive functioning in elderly men, women who use estrogen, and women who do not use estrogen. DESIGN: At two test times 18 months apart (time 1 and time 2), men (time 1, n = 31; time 2, n = 23), women who were using estrogen (time 1, n = 14; time 2, n = 10), and women who were not using estrogen (time 1, n = 41; time 2, n = 27), whose average age was 72.1 and 73.4 years at time 1 and time 2, respectively, were administered a battery of neuropsychological tests that measured verbal memory, visual memory, concentration and attention, language fluency, and semantic memory. Plasma levels of E2 and free T were assessed by radioimmunoassay. RESULTS: The men had higher free T levels than both groups of women at both test times. Although women who were using estrogen had higher E2 levels than those of the men and of the women who were not using estrogen, the men's E2 levels were also significantly higher than those of the women who were not using estrogen. Moreover, the women who were using estrogen and the men had higher Forward Digit Span scores compared with the women who were not using estrogen at both test times, and women who were using estrogen had higher Backward Digit Span scores than those who were not using estrogen. Both groups of women performed better than the men on the Category Retrieval Test (verbal fluency). The performance of women who were using estrogen on the Delayed Selective Reminding Test (long-term rote memory) improved over time compared with that of the men and of the women who were not using estrogen. CONCLUSIONS: These findings raise the possibility that higher E2 levels in elderly men and in women who use estrogen may protect against some declines in explicit memory with normal aging. PMID- 10810963 TI - Auditory brainstem response in postmenopausal women treated with hormone replacement therapy: a pilot study. AB - OBJECTIVE: To research the nongenital audiological target for gonadal steroids in postmenopausal women who are treated with hormone replacement therapy. DESIGN: Fifty postmenopausal volunteers were treated with hormone replacement therapy. Women with an intact uterus had sequential weekly transdermal estradiol plus nomegestrole acetate 5 mg orally for 12 days per month or a continuous daily oral dose of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg tablet. Eighteen surgically postmenopausal women received a weekly transdermal estradiol system. Twenty-five postmenopausal volunteers-5 with a natural menopause and 10 with a surgical menopause-and 20 premenopausal normally cycling women were used as a control group. Each woman performed auditory brainstem response by auditory evoked potentials for waves I, III, and V and for interpeak I-III, I-V, and III-V intervals. RESULTS: Women who were treated with hormone replacement therapy showed wave latencies and interpeak latencies shorter than those for postmenopausal women in the control group (p < or = 0.05), overlapping those of the premenopausal women (p > 0.05). Women who were treated with estrogen replacement therapy showed shorter time latencies than those treated with combined hormone replacement therapy (p < or = 0.05). CONCLUSIONS: Our data suggest that fluctuating hormone levels cause changes in auditory brain-stem response waves, even if the exact mechanism of activity of the gonadal steroids is not clear. However, we believe that estrogen may influence the neuronal plasticity, the metabolic levels of neurotransmitters, and thus the neuronal conduction time into the audiological system. PMID- 10810964 TI - Menopause across cultures: a review of the evidence. AB - OBJECTIVE: To review the cross-cultural evidence on menopausal symptoms to assess the extent of variability in symptomatology and the relative weight of hormonal and social factors. DESIGN: Literature review and critical summaries of available studies. RESULTS: Symptoms related to menopause are found in all regions of the world, although everywhere large proportions of women go through menopause uneventfully. The evidence does not support that women in developing countries report fewer symptoms than in industrialized countries. There is a great diversity in symptom frequencies across countries, and the association of symptoms with menopausal status is weak. A number of symptoms thought to be part of menopause are in fact not specific to it, although the evidence does support the narrow estrogen hypothesis of a core of symptoms associated with estrogen decline, namely vasomotor and vaginal symptoms. CONCLUSIONS: The association between hormonal changes and menopause symptomatology is complex and mediated by sociocultural factors. PMID- 10810965 TI - The medicalization of menopause: implications for recruitment of study participants. AB - OBJECTIVE: Early studies of menopause recruited sample populations from clinical settings; however, in the 1970s, populations drawn from health care settings were characterized as nonrepresentative because of symptom overreporting. This pilot study was carried out to test whether this characterization still holds true: that women who are drawn from clinical settings report more symptoms compared with women who are recruited from community and work sites. DESIGN: Open-ended interviews were carried out with patients aged 40-60 years in a physician's office (n = 50), in a chiropractor's office (n = 24), at two Breast Health Project sites (n = 50), and in several non-health care sites in the community (n = 81). Interviews were supplemented by anthropometrics and standardized return-by mail questionnaires. RESULTS: Women who experienced hot flashes and sweating were more likely to report having spoken with a physician about menopause. However, women who were drawn from the clinical setting were not significantly more likely to describe hot flashes, sweats, or mood changes and were significantly less likely to report headaches in relation to menopause compared with a community sample. Women who were drawn from the physician's office were more likely to use hormone replacement therapy and to have had a hysterectomy. CONCLUSIONS: This study suggests that because of the medicalization of menopause, we need to rethink our assumptions about the characteristics of populations drawn from health care settings. In western Massachusetts, place of recruitment did not predict symptom frequency. PMID- 10810966 TI - An electronic menstrual cycle calendar: comparison of data quality with a paper version. AB - OBJECTIVES: This pilot study compared a prototype electronic menstrual calendar on a handheld computer with a paper calendar for data quality and participants' perceptions. DESIGN: Twenty-three women completed identical information about menstrual bleeding and symptoms using paper and electronic calendars for 1 month each. RESULTS: Use of the paper calendar resulted in more missing data than the electronic calendar for bleeding characteristics (13% vs. 4%) and symptoms (35% vs. 4%). The electronic calendar's ability to log data entries revealed retrospective entry for 61% of the data. Total data entry and cleaning time was reduced by 81% with the electronic calendar. Overall, participants preferred the electronic (70%) to the paper (22%) calendar. CONCLUSIONS: Data quality with conventional paper calendars may be poorer than recognized. The data-logging feature, unique to the electronic calendar, is critical for assessing data quality. Electronic menstrual calendars can be useful data collection tools for research in women's health. PMID- 10810967 TI - Long-term results of Burch colposuspension. AB - BACKGROUND: We aimed to determine the long-term results of Burch colposuspension. METHODS: Patients who had undergone Burch colposuspension due to stress urinary incontinence (SUI) in our department between 1991 and 1995 were asked to participate in the study by telephone or mail. Fifty of 78 patients (64%) responded and these formed the study group. Patients were evaluated by a detailed questionnaire, pelvic examination, uroflowmetry and postvoid residual urine determination. Provocative stress test and urodynamic evaluations were performed in those who claimed leakage. Additionally, follow-up charts were retrospectively reviewed from the patients' files. RESULTS: Mean follow-up time was 50.6 months. The subjective cure rate was 52% and the surgical success rate was 84%. The patient satisfaction rate in terms of incontinence was 86%. No correlation was found between pre-operative patient characteristics (i.e. age, number of vaginal deliveries and pregnancies, menopause, previous anti-incontinence surgery and presence of detrusor instability) and outcome of surgery. Although no patient was performing clean intermittent catheterization in the long term, two patients had significant residual urine and obstructive flows. Three patients had severe pelvic prolapse that required surgical correction. CONCLUSIONS: Our results indicate that Burch colposuspension operation is an effective and durable choice of treatment with low complication rates for the treatment of SUI. PMID- 10810968 TI - Preclinical Cushing's syndrome: report of seven cases and a review of the literature. AB - BACKGROUND: Adrenal adenomas showing autonomous cortisol secretion without specific endocrine symptoms are sometimes discovered in patients with adrenal incidentalomas. This entity has been described as subclinical or preclinical Cushing's syndrome (PCS), but the endocrine data of reported cases have varied and the diagnostic criteria of PCS have been uncertain. METHODS: We report seven Japanese cases of PCS due to a unilateral, solitary adrenal adenoma with examination of the endocrine data of these patients. The diagnostic parameters of subtle hypercortisolism and the risk of postoperative adrenal insufficiency and surgical indications are discussed and reviewed. RESULTS: In the present cases, the most frequently found biochemical parameters of autonomous cortisol secretion were a low adrenocorticotropic hormone (ACTH) level (100%) and insufficient suppression of cortisol by low-dose dexamethasone (85.7%). Unilateral accumulation of radiopharmaceuticals in tumors was also frequently observed (100%). A postoperative hydrocortisone supplement was given to six of the seven patients for 5-122 days. It was not given to case 4, because a moderate response of 11-deoxycortisol to metyrapone was identified. Plasma ACTH levels and the diurnal rhythm of plasma cortisol rapidly recovered within 3 weeks postoperatively in six of the seven cases. CONCLUSION: This entity is heterogeneous and various degrees of cortisol excess have been observed. It should be diagnosed in the wide spectrum and the risk of adrenal insufficiency after surgery should be evaluated by dynamic tests such as the corticotropin releasing hormone (CRH) test. Based on the results of the present study and a review of the literature, PCS patients may not require hydrocortisone supplement therapy for a long period. PMID- 10810969 TI - Post-voiding repeated renal ultrasonography for slight hydronephrosis detected during screening for asymptomatic microhematuria. AB - BACKGROUND: There is not a well established method for further screening of asymptomatic microhematuria found through annual health examinations. However, it is apparent that a large number of examiners are using ultrasonography for screening. When slight hydronephrosis is found through screening, it is difficult to determine whether further examinations, such as intravenous pyelography (IVP), should be performed. In this paper, we discuss the usefulness of post-voiding repeated renal ultrasonography, which was applied to 57 patients with slight hydronephrosis found in a screening. METHODS: Ultrasonography in the kidney and the urinary bladder at the ultrasound laboratory of Toma Hospital was carried out on a total of 1906 patients with asymptomatic microhematuria, which was found by routine annual health examination, from January 1996 to December 1998. Slight hydronephrosis was found in 57 (14 males and 43 females) of 1906 patients and post-voiding repeated renal ultrasonography was carried out on all patients. RESULTS: Slight hydronephrosis had disappeared and IVP was avoided in 22 (38.6%) of 57 patients. Of 35 cases in which slight hydronephrosis persisted after urination, there were no abnormalities on IVP in 32 patients, while abnormalities were found in three patients. Two of these had ureteropelvic junction stenosis and the remaining one had a ureteral stone. CONCLUSION: The results of this study indicate that post-voiding repeated renal ultrasonography is useful for avoiding unnecessary further examination to determine whether upper urinary tract obstruction exists, when slight hydronephrosis is observed upon screening by simultaneous renal and urinary bladder ultrasonography for asymptomatic microhematuria. PMID- 10810970 TI - The correlation of stage and pathology of prostate cancer in Japan. AB - BACKGROUND: Few articles describe stage and pathology distribution of prostate cancer by age, yet understanding these topics may provide clues to the etiology of this disease. The present study describes the distribution of stage and pathology of prostate cancer in terms of age and considers effective detection of early stage cancer by mass screening (MS). METHODS: Data presented in this study were based on patient's files from MS and the Gunma Urological Oncology Study Group (GUOSG) registry from 1981 to 1996 in Gunma Prefecture, Japan. RESULTS: Age did not significantly correlate with stage distribution among the GUOSG patients, whereas stage tended to advance with age among the MS patients. Stage B was detected more frequently than any other stage in all age groups of the MS patients, whereas stage C and stage D were detected at similar rates in all age groups. After prostate specific antigen (PSA) had been introduced in MS, the percentage of early stage cancer increased at an earlier age. Pathology distribution did not significantly differ in any group. CONCLUSIONS: The data obtained from MS cancer patients showed that tumor stage tends to advance with age. We believe that the younger age group should be examined by MS using PSA to effectively detect early prostate cancer. PMID- 10810971 TI - Endoscopic neodymium:yttrium aluminium garnet (Nd:YAG) laser irradiation of a bladder hemangioma associated with Klippel-Weber syndrome. AB - A case of endoscopic neodymium:yttrium aluminium garnet (Nd:YAG) laser coagulation of a bladder hemangioma associated with Klippel-Weber syndrome is presented. The patient presented with extensive nevus and swelling of the left lower limb since birth. She was diagnosed with Klippel-Weber syndrome by angiography at the age of 1 year. Gross hematuria had been observed since she was 1 year old and, in addition, endoscopic examination revealed diffuse bladder hemangiomas. At 8 years of age, gross hematuria became worse and gait disturbance also appeared. She was referred to the Department of Urology at Tohoku University School of Medicine for endoscopic treatment in June 1998. Under general anesthesia, the bladder was inflated with CO2 gas and the hemangiomas were coagulated by Nd:YAG laser photonic irradiation endoscopically. Gross hematuria was markedly improved immediately following this treatment. Klippel-Weber syndrome is a rather uncommon disease which shows various types of vascular anomaly and hypertrophy of the lower extremities. Three to 6% of the patients have associated bladder hemangiomas. Although Nd:YAG laser irradiation provides results superior to alternative therapy and is the preferred treatment for most patients with bladder hemangioma, in this case, hemangiomas are multiple and present the possibility of re-bleeding, therefore long-term and close follow up is important. PMID- 10810972 TI - Optimization of orthotopic colonic bladder: implantation of tubularized mucosal flap from colonic segment into demucosalized urethra. AB - If the urethral mucosa of a female could be replaced by another mucosa, one could augment the indication for orthotopic reconstruction of the bladder. A mucosal replacement technique for the female urethra is reported. A colonic mucosal flap formed from a colonic bladder was used in a patient with bladder cancer for whom cystectomy was indicated. For 3 years after the surgery, the patient has maintained satisfactory voiding and urethral function. There has been no evidence of a recurrence. The orthotopic bladder with the mucosal replacement technique can be applied to female patients who have a bladder cancer with a high risk of urethral recurrence. PMID- 10810973 TI - Ruptured cerebral aneurysm not detected by magnetic resonance angiography in juvenile autosomal dominant polycystic kidney. AB - Recently, it has been reported that magnetic resonance angiography (MRA) is useful for screening and following up cerebral aneurysms in patients with autosomal dominant polycystic kidney disease (ADPKD). However, a patient was encountered with a ruptured cerebral aneurysm that was not detected by routine MRA. The patient, a 29-year-old man with ADPKD, was followed up at our hospital for more than 5 years. Ten months after an MRA examination, he suddenly developed severe headache. Brain computed tomography revealed subarachnoid hemorrhage. Digital subtraction angiography detected an aneurysm with a diameter of approximately 2 mm in the anterior communicating artery. Clipping of the aneurysm was immediately performed and he recovered without sequela after operation. Magnetic resonance angiography is useful to detect cerebral aneurysms, but it can not detect aneurysms measuring less than 4 mm. PMID- 10810974 TI - Pelvic congestion syndrome caused by inferior vena cava reflux. AB - We present a female patient who had pelvic congestion syndrome caused by inferior vena cava reflux associated with tricuspid regurgitation, but without other symptoms or signs related to her tricuspid regurgitation. Bladder pumping therapy was effective in improving her symptoms. PMID- 10810975 TI - Histology and tetracycline labeling of a single section of alveolar bone of first molars in the rat. AB - We observed the histology and tetracycline (TC) labeling in a single frontal section of alveolar bone of upper first molars of adolescent rats. A single injection of TC was administered intraperitonealy in adolescent rats. After three weeks, the upper jaws were immersed rapidly in liquid nitrogen and sectioned. Five micrometer unfixed, undecalcified frozen sections were cut and observed by light and fluorescence microscopy. Frontal sections of the upper first molar area revealed that the structural relationships among the roots, the periodontal ligament and the alveolar bone, and also between the cervical enamel and the attachment epithelium were well preserved. The TC labeling lines in the sections were very clear and distinguished new bone from old bone. The brightness of the lines differed among regions. An analysis of the brightness in the same section suggested a difference in the bone forming activity at the time of injection. PMID- 10810976 TI - Evaluation of LR white resin for histology of the undecalcified rat tibia. AB - Histology of plastic embedded undecalcified bone represents a challenging problem to the histotechnologist. We outline here an exploration of LR White resin as a suitable medium for histologic study of undecalcified rat tibia. A procedure was developed for light microscopy of rat tibia embedded in LR White and sectioned by sawing-grinding technics. The specimens were fixed in 10% neutral buffered formalin or alcohol-acetic acid-formol, dehydrated in ethanol, defatted in chloroform followed by resin infiltration and heat-curing of embedded blocks. The procedure of dehydration, defatting, infiltration, and polymerization can be completed within 10 days. Cold curing with accelerator provided by the manufacturer did not yield superior results compared to blocks cured with heat. Thick sections were obtained using a diamond wire saw, attached to plexiform slides, then ground and polished. Surface staining with Von Kossa silver reagent or toluidine blue revealed satisfactory morphological preservation of the mineralized bone sections. Artifacts like small bubbles appeared occasionally and could not be avoided despite prolonged infiltration or cold curing of blocks. Our method is relatively simple for base-line histologic study of rat tibia. The method offers advantages such as easy adaptability, reliable stainability, contrast, and resolution of bone architecture and marrow cells. Two other embedding media, Micro-Bed resin and Unicryl, were also tested, but produced inferior results. PMID- 10810977 TI - Comparative image analysis of EGF immunoreaction in rat submandibular gland using 3,3'-diaminobenzidine with metal enhancer substrate. AB - We have shown the efficacy of image analysis using 3,3'-diaminobenzidine (DAB) with a metal enhancer substrate for demonstrating quantitative differences in the amount of epidermal growth factor (EGF) in the submandibular gland from normal, castrated, and testosterone propionate (TP) treated castrated rats. Immunohistochemical determination of EGF visualized by DAB-nickel reagent was performed with image analysis using a computed image analyzer system (ACAS 570). Immunohistochemistry for EGF disclosed positive staining in granular convoluted tubule cells in the tissue sections from each experimental group. Using the tools of a competent data program installed in the ACAS 570 software, we measured quantitative differences among the experimental glands examined. Castration was shown to elicit a significant reduction in the EGF-positive area and staining intensity, and administration of TP to the castrated animals restored these parameters to levels greater than those of normal rats. Our study demonstrates that a simple, inexpensive, commercially available metal enhancer substrate can be applied accurately to the computer assisted quantification of histochemical hormone-induction studies. PMID- 10810978 TI - Formaldehyde prepared from paraformaldehyde is stable. AB - For critical histological investigations, tissue fixation is sometimes carried out in formaldehyde freshly prepared from paraformaldehyde by heating. The purity of formaldehyde produced in this way is superior to that of commercial stock solutions. We studied the stability of freshly prepared formaldehyde solutions by determination of pH and titration of acid, which reflect the formation of formic acid. It was found that very small amounts of acid are produced during the heating of paraformaldehyde. Prolonged heating or storage of freshly prepared formaldehyde for up to 8 days did not significantly increase the amount of acid. It was also found that heating of the paraformaldehyde is not necessary, since depolymerization may take place at room temperature. We conclude that formaldehyde prepared from paraformaldehyde remains stable for considerable periods of time, and it is therefore unnecessary to prepare it immediately prior to fixation. Also, in many cases, buffering of the fixative may be omitted, since only minor changes in the pH occur during fixation. PMID- 10810979 TI - Embedding thin plant specimens for oriented sectioning. AB - Small plant structures such as small primary roots, filamentous mosses and algae are difficult to orient for sectioning since they become wavy and curl during embedding. A method is described for embedding and orienting tiny plant specimens in a glycol methacrylate resin using self-constructed flat molds. Prior to sectioning, small samples can be oriented in both the longitudinal and the transverse plane. As several samples can be sectioned simultaneously, time consuming trimming of the blocks is reduced substantially. The efficiency of this technique has been demonstrated using the tiny roots of the model plant Arabidopsis thaliana (L.) Heynh. PMID- 10810980 TI - Use of a Technovit 7200 VLC to facilitate integrated determination of aluminum by light and electron microscopy. AB - Technovit 7200 VLC is an excellent embedding medium for both inorganic histochemistry by light microscopy and X-ray microanalysis by scanning and transmission electron microscopy. Liver samples from rats after intraperitoneal treatment with aluminum chloride were fixed in glutaraldehyde and embedded in the resin. Thick sections were easily cut on an ultramicrotome and stained with aluminon for aluminum (Al). An intense positive reaction with aluminon was observed in the Kupffer cells by light microscopy. The surface structures of the same resin block cut for light microscopy were observed under a scanning electron microscope fitted with an energy dispersive X-ray spectrometer. The Kupffer cells appeared white in the backscattered mode. Localization of Al in the Kupffer cells was confirmed by an X-ray distribution map in the scanning electron microscope. Subcellular localization of Al in the Kupffer cells was performed on the same semithin sections using a transmission electron microscope equipped with an energy dispersive X-ray spectrometer. Most Al was found in lysosomes of the Kupffer cells. The resin was stable in the electron beam and chlorine-free. PMID- 10810982 TI - Hexamethyldisilazane for scanning electron microscopy of Gastrotricha. AB - We evaluated treatment with hexamethyldisilazane (HMDS) as an alternative to critical-point drying (CPD) for preparing microscopic Gastrotricha for scanning electron microscopy (SEM). We prepared large marine (2 mm) and small freshwater (100 microm) gastrotrichs using HMDS as the primary dehydration solvent and compared the results to earlier investigations using CPD. The results of HMDS dehydration are similar to or better than CPD for resolution of two important taxonomic features: cuticular ornamentation and patterns of ciliation. The body wall of both sculpted (Lepidodermella) and smooth (Dolichodasys) gastrotrichs retained excellent morphology as did the delicate sensory and locomotory cilia. The only unfavorable result of HMDS dehydration was an occasional coagulation of gold residue when the solvent had not fully evaporated before sputter-coating. We consider HMDS an effective alternative for preparing of gastrotrichs for SEM because it saves time and expense compared to CPD. PMID- 10810981 TI - A modification of Mayer's tannic acid-ferric chloride staining method for demonstrating cellular membranous systems for light microscopy. AB - To observe cellular membranous systems under a light microscope, we modified Mayer's tannic acid-ferric chloride stain method by adding a treatment with hematoxylin after the original procedure. We used the modified tannic acid-ferric chloride (MTA-Fe) stain method to examine kidneys, liver, heart, trachea, epididymides and other organs of rats and dogs. The MTA-Fe stain clearly demonstrated the basement membrane, brush border, basolateral invaginations and cell processes in the kidneys which enabled easy differentiation of the S1 and S3 segments of proximal convoluted tubules. Our technique also demonstrated hepatic cell membranes and bile canaliculi in the liver, cross striations and longitudinal traveling of myofibrils in the heart, cilia of the epithelial cells in the trachea, and stereocilia and terminal bars in the epididymis. The MTA-Fe stain is a convenient method to visualize cellular membranous systems even for light microscopy. The stain has the advantages of using no toxic materials, simple and easy technique, little variation of staining results, and little fading for several months after staining. PMID- 10810983 TI - Diaminobenzidine as a myelin stain in semithin plastic sections. PMID- 10810984 TI - Virchow's contributions to veterinary medicine: celebrated then, forgotten now. AB - In 1858, Rudolf Virchow, the professor of pathology in Berlin University, published the book "Cellular Pathology". A compendium of his lectures to physicians and medical students, he introduced the use of microscopy for the study of human diseases. To an astonishing extent Rudolf Virchow was helpful to the disciplines of veterinary medicine (and veterinary pathology). Considered a scientific genius in several disciplines, this essay deals exclusively with the devotion of Virchow, a scholarly physician, to the profession of veterinary medicine. He respected veterinary research, supported governmental veterinary education, and provided a role model for the veterinarians who were drafting control legislation of contagious diseases in livestock. Repeatedly, he responded in help when seemingly irretrievable problems arose. Examples of Virchow's activities in the realms of veterinary medicine and pathology are marshalled here to shed light on this pioneer "veterinary pathologist". In celebration of 50 years of the American College of Veterinary Pathologists in 1999, it is timely to remember that Rudolf Virchow, the father of cellular pathology, also fathered veterinary pathology, whose offsprings in Canada and the U.S.A. (Osler, Clement, Williams, Olafson, Jones) had enabled them to form and foster the A.C.V.P. PMID- 10810985 TI - Pathology of fatal West Nile virus infections in native and exotic birds during the 1999 outbreak in New York City, New York. AB - West Nile fever caused fatal disease in humans, horses, and birds in the northeastern United States during 1999. We studied birds from two wildlife facilities in New York City, New York, that died or were euthanatized and were suspected to have West Nile virus infections. Using standard histologic and ultrastructural methods, virus isolation, immunohistochemistry, in situ hybridization and reverse-transcriptase polymerase chain reaction, we identified West Nile virus as the cause of clinical disease, severe pathologic changes, and death in 27 birds representing eight orders and 14 species. Virus was detected in 23/26 brains (88%), 24/ 25 hearts (96%), 15/18 spleens (83%), 14/20 livers (70%), 20/20 kidneys (100%), 10/13 adrenals (77%), 13/ 14 intestines (93%), 10/12 pancreata (83%), 5/12 lungs (42%), and 4/8 ovaries (50%) by one or more methods. Cellular targets included neurons and glial cells in the brain, spinal cord, and peripheral ganglia; myocardial fibers; macrophages and blood monocytes; renal tubular epithelium; adrenal cortical cells; pancreatic acinar cells and islet cells; intestinal crypt epithelium; oocytes; and fibroblasts and smooth muscle cells. Purkinje cells were especially targeted, except in crows and magpies. Gross hemorrhage of the brain, splenomegaly, meningoencephalitis, and myocarditis were the most prominent lesions. Immunohistochemistry was an efficient and reliable method for identifying infected cases, but the polyclonal antibody cross reacted with St. Louis encephalitis virus and other flaviviruses. In contrast, the in situ hybridization probe pWNV-E (WN-USAMRIID99) reacted only with West Nile virus. These methods should aid diagnosticians faced with the emergence of West Nile virus in the United States. PMID- 10810986 TI - Skeletal lesions of canine hepatozoonosis caused by Hepatozoon americanum. AB - Canine hepatozoonosis, caused by Hepatozoon americanum, is an emerging tick-borne disease of dogs in North America. In addition to the skeletal and cardiac myositis that are prominent features of the disease, there is disseminated periosteal bone proliferation in most dogs that manifest clinical disease. Each of six experimentally infected animals (four dogs and two coyotes) and seven of eight naturally infected dogs had gross or histopathologic osteoproliferative lesions. Experimental animals were 6-9 months of age when exposed. Naturally infected dogs were 8 months to 11 years old when subjected to necropsy. Lesions occurred primarily on the diaphysis of the more proximal long bones of the limbs; however, flat and irregular bones were frequently involved. Lesions involving metacarpals, metatarsals, and digits were infrequent. The earliest observed periosteal lesions were in an experimentally infected dog 32 days after exposure to sporulated oocysts of H. americanum. There were hypertrophy and hyperplasia of osteoprogenitor cells, and osteoblasts appeared in the cellular zone of the periosteum. Spicules of woven bone oriented perpendicularly to bone cortex followed. Later yet, periosteal new bone was remodeled and tended to become oriented parallel to the cortical bone. Horizontally oriented zones of remodeled, condensed bone sometimes occurred in multiple layers on the original cortex, forming "pseudocortices." The osseous lesions of American canine hepatozoonosis, with few variations, are remarkably similar to those of hypertrophic osteopathy in domestic dogs and other mammalian species, including humans. PMID- 10810987 TI - Reduction of carbon tetrachloride-induced hepatotoxic effects by oral administration of betaine in male Han-Wistar rats: a morphometric histological study. AB - Eighty-five male Han-Wistar rats were arranged into three groups: CCl4-exposed rats, CCl4 + betaine-exposed rats, and control rats. To see the effect of betaine alone, five rats of the control and of the CCl4 + betaine groups were sacrificed after 7 days, before exposure to CCl4. After that, two of the groups (the CCl4 and CCl4 + betaine groups) were exposed to CCl4 (1 ml/kg per day subcutaneously [SC] for 4 consecutive days), and one of the groups (control group) was given olive oil (1 ml/kg per day SC for 4 consecutive days). At the start of the study (day 0), day 1, day 2, day 3, day 4, and 3 days after the last CCl4 and olive oil injections (day 7), samples of five rats per group were sacrificed, and the livers were taken for chemical analyses and histological examination. Oral betaine, after the acclimation period of a week, increased the number of mitochondria but not mitochondria size (day 0), compared with the case in control rats. Exposure to CCl4 resulted in centrilobular hepatic steatosis, and the administration of betaine significantly reduced this. Morphometric analyses also revealed that the addition of betaine increased the volume density of rough endoplasmic reticulum (RER) in the perinuclear areas of liver cell cytoplasm (day 7). Additionally, the administration of betaine prevented the reduction of Golgi complexes and mitochondrial figures in the cytoplasm observed after the exposures to CCl4. Also, the volume density of mitochondria was smallest in the CCl4-group, but the difference was not statistically significant. The results indicate that oral betaine either improves recovery or reduces the toxic effects of CCl4 on cell organelles in liver cells of male Han-Wistar rats. PMID- 10810988 TI - Immunohistologic detection of estrogen receptor alpha in canine mammary tumors: clinical and pathologic associations and prognostic significance. AB - Eighty-nine canine mammary tumors and dysplasias of 66 bitches were investigated to determine the immunohistochemical expression of classical estrogen receptor (ER-alpha) and its clinical and pathologic associations and prognostic value. A complete clinical examination was performed and reproductive history was evaluated. After surgery, all animals were followed-up for 18 months, with clinical examinations every 3-4 months. ER-alpha expression was higher in tumors of genitally intact and young bitches (P < 0.01, P < 0.01) and in animals with regular estrous periods (P = 0.03). Malignant tumors of the bitches with a previous clinical history of pseudopregnancy expressed significantly more ER alpha (P = 0.04). Immunoexpression of ER-alpha decreased significantly with tumor size (P = 0.05) and skin ulceration (P = 0.01). Low levels of ER-alpha were significantly associated with lymph node involvement (P < 0.01). Malignant tumors had lower ER-alpha expression than did benign tumors (P < 0.01). Proliferation index measured by proliferating cell nuclear antigen immunostaining was inversely correlated with ER-alpha scores (P = 0.05) in all tumors. Low ER-alpha levels in primary malignant tumors were significantly associated with the occurrence of metastases in the follow-up (P = 0.03). Multivariate analyses were performed to determine the prognostic significance of some follow-up variables. ER-alpha value, Ki-67 index, and age were independent factors that could predict disease free survival. Lymph node status, age, and ER-alpha index were independent prognostic factors for the overall survival. The immunohistochemical detection of ER-alpha in canine mammary tumors is a simple technique with prognostic value that could be useful in selecting appropriate hormonal therapy. PMID- 10810989 TI - p53 gene mutations occurring in spontaneous benign and malignant mammary tumors of the dog. AB - Sixty-three cases of benign and malignant canine mammary tumors were analyzed to define the alteration of exons 5-8 for the p53 tumor suppressor gene using polymerase chain reaction direct sequence analysis with paraffin-embedded tissues. Four missense mutations were found in 38 benign mammary tumors (11%), and five missense (one tumor had two missense mutations) and one nonsense mutations were found in 25 mammary carcinomas (20%). These data suggest that the p53 gene alterations might be initiated at an early stage of canine mammary carcinogenesis and p53 mutations might be associated with malignancy. However, there was no evidence of any relationship between the p53 alterations and the histologic types of tumors or breeds of dogs. PMID- 10810990 TI - Viral wasting syndrome of swine: experimental reproduction of postweaning multisystemic wasting syndrome in gnotobiotic swine by coinfection with porcine circovirus 2 and porcine parvovirus. AB - One-day-old gnotobiotic piglets were inoculated intranasally with in vitro passaged porcine circovirus 1 (PCV-1), PCV-2, and porcine parvovirus (PPV) alone or in combination (PCV-1/PCV-2, PCV-1/PPV, and PCV-2/PPV). Piglets were evaluated for 1) the development of porcine postweaning multisystemic wasting syndrome (PMWS), 2) distribution of viral antigens by immunochemistry, and 3) viremia and the presence of viral DNA in nasal and ocular secretions and feces. All single agent-infected piglets and piglets infected with PCV-1/PCV-2 or PCV-1/PPV were clinically asymptomatic. They were transiently viremic and seroconverted to homologous virus(es). At termination of the study on postinfection day (PID) 35, microscopic lesions were restricted to focal inflammatory cell infiltrates in livers and myocardia. One piglet given PCV-1/PPV was PPV viremic for 2 weeks after infection and had lymphangiectasia of the spiral and descending colon associated with granulomatous inflammation. All four PCV-2/PPV-inoculated piglets developed PMWS, characterized by sudden onset of depression and anorexia, icterus, and submucosal edema. One piglet became moribund on PID 27, and the remaining three piglets were euthanatized between PID 27 and PID 30 because of severe disease. Lymph nodes were small and the livers were mottled. Disseminated angiocentric granulomatous inflammation was present in all tissues examined except the brain. Multiple lightly basophilic intracytoplasmic inclusion bodies were identified in macrophages and histiocytes. PCV-2 antigen was widely distributed within macrophages; PPV antigen was sparse. Hepatocellular necrosis and bile retention were prominent. PCV-2 DNA was identified in ocular, fecal, and nasal secretions. Terminal sera contained antibodies to PPV (4/4) and PCV-2 (3/ 4). Production of PMWS in gnotobiotic swine appears to require PCV-2 and additional infectious agents such as PPV for full disease expression in gnotobiotic piglets. PMID- 10810991 TI - Anophthalmia and retinal degeneration associated with stenosis of the optic foramen in Fischer 344 rats. AB - Bilateral anophthalmia was discovered in a male rat (No. 1) and unilateral anophthalmia was found in the left eye of two female rats (Nos. 2 and 3) derived from a Fischer 344 inbred colony. One male rat (No. 4), a littermate of No. 3, had externally normal eyes, but his left eye had severe retinal atrophy, which was detected by ophthalmoscopy. The eyelids in both eyes of No. 1 and in the left eyes of Nos. 2 and 3 were present. Radiography of the skull revealed small optic foramina on both sides of No. 1 and on the affected side of Nos. 2, 3, and 4. Histologically, both globes and optic nerves (ONs) of No. 1 and the left globe and ONs of Nos. 2 and 3 were completely missing. Diffuse retinal degeneration in the left globe and axonal degeneration in the left ON and the right optic tract were observed in No. 4. Dysplasia of the sphenoid bone associated with stenosis of the optic foramen was detected on the affected side of all rats. Thus, ON aplasia in anophthalmic rats and atrophic ON in a rat with retinal degeneration seem to be closely related to stenosis of the optic foramen. PMID- 10810992 TI - Immunohistopathologic characterization of a dermal melanocytoma-acanthoma in a German Shepherd Dog. AB - A cutaneous melanocytoma-acanthoma in a 2-year-old female German Shepherd Dog was characterized by the presence of two populations of neoplastic cells: epithelial and melanocytic. The epithelial component consisted of nests of well differentiated stratified squamous epithelium closely associated with neoplastic melanocytes. The epithelial cells immunoreacted with both monoclonal and polyclonal anti-cytokeratin antibodies, and immunoreaction to S-100 protein and vimentin was observed in the melanocytic cells. This rare pigmented skin neoplasm of the dog apparently has a benign behavior. PMID- 10810993 TI - Cerebello-olivary and lateral (accessory) cuneate degeneration in a juvenile American Miniature horse. AB - A 12-month-old American Miniature horse colt was presented to the Virginia Tech Veterinary Teaching Hospital with a 7-month history of progressive ataxia. Physical examination revealed a head intention tremor, base-wide stance, and ataxia. Necropsy findings were confined to the brain. There were bilateral areas of liquefactive necrosis and cavitation corresponding to the dorsal accessory olivary and lateral (accessory) cuneate nuclei. Cerebellar folia of the dorsal vermis were thin. Microscopically, the cerebellar cortex was characterized by patchy areas of Purkinje cell loss with associated variable thinning of the molecular and granule cell layers and astrogliosis. Dorsal accessory olivary and lateral cuneate nuclei were cavitated and had mild glial response around their periphery. Additionally, a focus of necrosis and neuropil vacuolization was found in the right putamen. These findings indicate the presence of a neurodegenerative disorder centered, but not confined to, the cerebellum and its connections in this American Miniature horse colt. PMID- 10810994 TI - A subependymal giant cell astrocytoma in a cat. AB - A 6-year-old spayed female Domestic Shorthair cat presented with a 1 to 2-month history of blindness and altered behavior. At necropsy, a 1-cm-diameter, firm white mass was found arising from the subependymal region of the right lateral ventricular wall that protruded into and partially filled the lumen. Histologically, there was a well-demarcated, expansile paraventricular neoplasm composed of moderately pleomorphic cells within a richly fibrillar matrix arranged in interlacing streams and perivascular pseudorosette-like patterns. Neoplastic cells varied in morphology from small spindloid cells to larger polygonal cells with eccentric vesicular nuclei to neuronlike cells with vesicular nuclei and prominent nucleoli. The mitotic index was low. Immunohistochemically, neoplastic cells were positive for S-100 protein, glial fibrillary acidic protein, and neuron-specific enolase and negative for neurofilament protein. Ultrastructurally, the cells contained few to abundant bundles of intermediate filaments with variable numbers of mitochondria, endoplasmic reticulum, and ribosomes. These features are characteristic of subependymal giant cell astrocytoma (SEGA) in humans. To our knowledge, this is the first reported case of SEGA in domestic animals. PMID- 10810995 TI - C-cell carcinoma (medullary thyroid carcinoma) associated with multiple endocrine neoplasms in a ferret (Mustela putorius). AB - A firm, infiltrative mass was found in the thyroid region of an adult castrated male ferret (Mustela putorius) presenting with vague signs of weight loss, minor inappetence, and decreased activity. Efforts to surgically excise the tissue were unsuccessful, and the animal was euthanatized. Gross and histopathologic evaluation revealed multiple endocrine neoplasms, including C-cell carcinoma, adrenocortical adenoma, pheochromocytoma, and endocrine tumor of the pancreas. This is the first descriptive account of a C-cell carcinoma, also known as medullary thyroid carcinoma, in a ferret, although other endocrine neoplasms in this species have been reported with some frequency. These findings mimic features observed in human multiple endocrine neoplasia syndromes. PMID- 10810996 TI - Enterococcus hirae enteropathy with ascending cholangitis and pancreatitis in a kitten. AB - A 2-month-old female Persian cat that had been showing episodes of anorexia and diarrhea for the previous 4 weeks was presented in shock and died 2 days later. Numerous Gram-positive cocci were located along the brush border of small intestinal villi, without significant inflammatory infiltration. Similar bacteria were present within hepatic bile ducts and pancreatic ducts and were associated with suppurative inflammation and exfoliation of epithelial cells. Culture of the liver and lung yielded bacteria identified as Enterococcus hirae. Fecal culture from an asymptomatic adult female from the same cattery also yielded large numbers of E. hirae. To our knowledge, this is the first report of E. hirae enteropathy in a cat and the first report of ascending cholangitis and ductal pancreatitis caused by an Enterococcus spp. PMID- 10810997 TI - The role of inflammation in vascular diseases. AB - When the body responds to an infectious insult, it initiates an immune response to eliminate the pathogen. The hallmark of the immune response is an inflammatory cascade that can also do extensive damage to host tissues. Inflammation is a major contributing factor to many vascular events, including atherosclerotic plaque development and rupture, aortic aneurysm formation, angiogenesis, and ischemia/reperfusion damage. The immune response is mediated by both circulating and resident leukocytes and the cells with which they interact (e.g., vascular endothelium and smooth muscle cells). The process is orchestrated by the activity of a changing series of released and displayed mediators. These include the expression of adhesion molecules on leukocytes and underlying vascular endothelium and the release of cytokines, chemokines, and tissue-destructive metalloproteases and reactive oxygen species. This review focuses on the causes, the inflammatory processes involved, and possible strategies for decreasing vascular disease through regulation of the inflammatory response. PMID- 10810998 TI - Definition of human blood monocytes. AB - At the Definition of Human Blood Monocytes conference held in Munich, Germany in October 1999, experts met to discuss data on the characterization of monocytes in health and disease. Emphasis was on how to best define these cells, on which subpopulations can be identified, and on how the monocytes can best be distinguished from other cells such as natural killer cells, granulocytes, and dendritic cell precursors. PMID- 10810999 TI - The critical role of CD40/CD40L in the CD4-dependent generation of CD8+ T cell immunity. AB - Control of virus infections and eradication of tumors usually involves the lytic activity of CD8+ cytotoxic T lymphocytes (CTLs). The induction of effective CTL immunity relies on several factors, one of the most important of which is CD4+ T cell help. Numerous studies have demonstrated the dependence of CTL priming on the presence of CD4+ T cells, but until recently little was known of the mechanisms regulating this process. Based on reports that CD4+ and CD8+ T cells must recognize antigen on the same antigen-presenting cell (APC), help was originally thought to be provided through the delivery of short-range, CD4+ T cell-secreted cytokines. However, the results of subsequent studies favor an alternative mechanism, whereby CD4+ T cells modify the APC, converting it into a stimulatory cell for CD8+ T cell priming. It is important that CD40 and its ligand, CD40L, have been implicated in the provision of this help and, in particular, the generation of long-lasting CTL memory. PMID- 10811000 TI - Induction of cyclooxygenase-2 by human monocytes exposed to group B streptococci. AB - Group B streptococcal (GBS) infections are associated with high morbidity and mortality. The molecular pathways mediating the pathophysiological events in GBS infection are not fully delineated. Cyclooxygenases (COX) are the enzymes that convert arachidonate to active eicosanoids. To identify the effects of GBS on eicosanoid metabolism and regulatory mechanisms, we exposed human monocytes to GBS and found that they secreted prostaglandin E2, prostacyclin, and thromboxane A2. Exposure to GBS caused monocytes to express COX-2 mRNA and protein in both a time- and concentration-dependent manner that correlated with eicosanoid production. COX-1 protein was unchanged. Addition of the anti-inflammatory cytokines interleukin (IL)-4 or IL-10 markedly attenuated GBS-induced COX-2 protein accumulation after GBS exposure, as did inhibition of p38 MAPK. Our experiments are the first to show that exposure of monocytes to a gram-positive bacterium (GBS) results in induction of functional COX-2, suggesting that eicosanoids may play important roles in the pathogenesis of GBS infections. PMID- 10811001 TI - Migration of dendritic cells within 3-D collagen lattices is dependent on tissue origin, state of maturation, and matrix structure and is maintained by proinflammatory cytokines. AB - The function of dendritic cells (DC) depends on active migration through three dimensional (3-D) extracellular matrices. We have analyzed the migration of murine DC from different tissue origins within 3-D collagen lattices through the use of time-lapse videomicroscopy and single-cell tracking. Directly after incorporation, 50-90% of DC from the spleen (spDC) and Langerhans cells freshly isolated from the epidermis (fLC) displayed active motility in these matrices. Whereas mature spDC showed multilateral pseudopod dynamics as well as fast and heterogeneous migration, immature fLC displayed a spherical shape with faint membrane processes and very homogenous, slow migration characteristics. In the absence of external stimuli, migration of both, spDC and fLC, vanished after >36 h due to cell death. Maintaining fLC viability by external granulocyte-macrophage colony-stimulating factor or tumor necrosis factor alpha prolonged migration up to 5 days. During this period fLC transformed into mature cells with large dendrites, thereby developing a heterogeneous migration pattern more similar to spDC. In randomly polymerized collagen matrices cell paths were without preferential orientation. In contrast, in artificially aligned lattices directional paths in accordance with the forced fiber orientation were observed. Thus, migration is an inherent property of DC, largely influenced by tissue origin, degree of maturity, and the 3-D structure of the environment. PMID- 10811002 TI - Hydrogen peroxide-induced phospholipase D2 activation in lymphocytic leukemic L1210 cells. AB - Extracellular hydrogen peroxide (H2O2) has been implicated in the activation of phospholipase D (PLD). However, it was still unclear how this activation occurs and what the molecular identity of the H2O2-stimulated PLD isozyme is. This study shows that H2O2 potently increases the PLD activity in mouse lymphocytic leukemic L1210 cells, which contain exclusively PLD2. In addition, H2O2 increased PLD activity only in PLD2-transfected COS-7 cells and not in PLD1-transfected cells. This suggests that PLD2 is selectively activated by H2O2. Depletion of extracellular Ca2+ with EGTA completely blocked the H2O2-induced PLD activation, indicating that Ca2+ influx is required. Moreover, pretreatment of the cells with the protein kinase C (PKC) inhibitors GF-109203X and RO-31-8220 and down regulation of PKCalpha by prolonged treatment with 4beta-phorbol 12-myristate 13 acetate inhibited the H2O2-stimulated PLD2 activity, which points to the involvement of PKCalpha. Based on these new findings we suggest that PLD2 activity is specifically up-regulated by H2O2 and that the H2O2-induced PLD2 activation is mediated by Ca2+ influx and PKCalpha activation. PMID- 10811003 TI - Diazepam-binding inhibitor-derived peptides induce intracellular calcium changes and modulate human neutrophil function. AB - We studied the effects of two diazepam-binding inhibitor (DBI)-derived peptides, triakontatetraneuropeptide (DBI 17-50, TTN) and eiksoneuropeptide (DBI 51-70, ENP), on cytosolic free Ca2+ concentrations ([Ca2+]i), chemotaxis, superoxide anion (O2-) generation, and phagocytosis in human neutrophils. Both TTN and ENP induced a rapid and transient rise of [Ca2+]i. The effect of TTN depended on the presence of extracellular Ca2+, whereas the effect of ENP also persisted after extracellular Ca2+ chelation. TTN induced neutrophil chemotaxis, stimulated O2- generation, and enhanced phagocytosis. ENP did not affect cell migration and oxidative metabolism but enhanced phagocytosis. Both peptides modulated N-formyl methionyl-leucyl-phenylalanine- and phorbol myristate acetate-induced O2- generation. Because neutrophils express benzodiazepine receptors of the peripheral type (pBRs) and DBI-derived peptides may interact with such receptors, we investigated the possible role of pBRs in TTN- or ENP-induced effects. The synthetic pBR ligand RO 5-4864 increased [Ca2+]i through extracellular Ca2+ influx and this effect was prevented by the pBR antagonist PK-11195. RO 5-4864, however, was ineffective on neutrophil migration and O2- generation and only slightly affected phagocytosis. Moreover, PK-11195 delayed the [Ca2+]i rise induced by TTN but did not significantly affect its extent, and had no effect on the [Ca2+]i rise induced by ENP. We conclude that DBI-derived peptides induce [Ca2+]i changes and modulate neutrophil function mainly through pBR-independent pathways. In view of the wide cell and tissue distribution of DBI in the brain and in peripheral organs, modulation of neutrophil function by DBI-derived peptides may be relevant for both the neuroimmune network and the development and regulation of the inflammatory processes. PMID- 10811004 TI - Insights into the role of gammadelta T lymphocytes in the immunopathogenic response to thermal injury. AB - Studies have shown that cell-mediated immunity is markedly suppressed after thermal injury. T lymphocyte dysfunction and macrophage hyperactivity have been implicated as causative factors. Previous studies have primarily examined the effects of thermal injury on alphabeta T lymphocytes; however, the role of gammadelta T lymphocytes in the immune response after thermal injury is unclear. Therefore, wild-type mice and mice lacking the TCR delta gene (TCR delta-/-) were subjected to a third-degree scald burn and cell-mediated immune responses assessed at 7 days post-injury. TCR delta-/- mice had 75% mortality after burn injury compared with 25% mortality in the wild-type group. Plasma interleukin-6 (IL-6) levels were significantly elevated at 2, 4, and 18 h post-injury, whereas no difference was observed in tumor necrosis factor alpha (TNF-alpha) and prostaglandin E2 (PGE2) plasma levels. Plasma levels of these inflammatory mediators were similar in wild-type and TCR delta-/- mice post-injury. Splenic macrophage PGE2, IL-6, TNF-alpha, and IL-10 production was significantly increased in wild-type mice at 7 days post-injury, whereas macrophages from injured TCR delta-/- mice had a significantly attenuated capacity to produce IL-6 and TNF-alpha. In contrast, the increased release of PGE2 and IL-10 by macrophages post-injury was not reduced in TCR delta-/- mice. These results implicate a dual role for gammadelta T lymphocytes in the immunopathogenic response to burn injury: (1) they contribute to survival from the insult; and (2) they mediate the induction of a pro-inflammatory macrophage phenotype at 7 days post-injury. Thus, gammadelta T lymphocytes, in part through the modulation of macrophage activity, appear to contribute to the immune dysfunction after thermal injury. PMID- 10811005 TI - Neutrophil chemotaxis in moving gradients of fMLP. AB - In this study the fluid gradient chamber, a modified version of the Boyden chamber that enables mobile gradients, was used to study the migration of human granulocytes in gradients of fMLP. Temporal chemotactic gradients were created by moving density-stabilized spatial gradients at different velocities in relation to migrating cells. Random and directed cell migration was quantified by applying a theoretical population distribution model to experimental cell distributions obtained from cell counts at different depths in the filters. Rates of random and directed migration generally increased with gradient velocity. At negative gradient velocities, i.e., when the gradients were moved in a direction opposite to that of cell migration to decrease fMLP concentration over time, random and directed migration was inhibited. At positive gradient velocities, migration rates were not significantly different from those seen in immobile gradients. The fact that the rate of directed migration was smaller at negative gradient velocities indicates that negative temporal gradients reduced the average speed and/or orientation of the chemotactically migrating cells. In immobile gradients, the cells generated a small concentration increase over time when they migrated in the up-gradient direction. Consequently, a positive temporal gradient as perceived by the cells may act as a positive feedback signal to maintain chemotactic migration. PMID- 10811006 TI - Inflammatory neutrophils retain susceptibility to apoptosis mediated via the Fas death receptor. AB - Apoptosis and clearance of neutrophils is essential for successful resolution of inflammation. Altered signaling via the Fas receptor could explain the observed prolongation of neutrophil lifespan and associated tissue injury at inflammatory sites. We therefore compared inflammatory neutrophils extracted from joints of rheumatoid arthritis patients, with peripheral blood neutrophils. Inflammatory neutrophils underwent constitutive apoptosis in culture more rapidly than peripheral blood neutrophils; this was not explained by changes in surface expression of Fas or by induction of Fas ligand. Inflammatory neutrophils remained sensitive to Fas-induced death, at levels comparable to those seen in peripheral blood neutrophils. Similarly, granulocyte-macrophage colony stimulating factor reduced apoptosis but did not abolish signaling via Fas. These data provide evidence for the rate of apoptosis in inflammatory neutrophils being continually modulated by death and survival signals in the inflammatory milieu. This allows for rapid resolution of inflammation as levels of survival factors fall, and suggests new strategies for inducing resolution of inflammation. PMID- 10811007 TI - Calcitonin-gene related peptide (CGRP) inhibits interleukin-7-induced pre-B cell colony formation. AB - Calcitonin gene-related peptide (CGRP) is a sensory neuropeptide with inflammatory and immunoregulatory activities. Its role in B lymphocyte development was investigated using a pre-B cell colony-forming assay. Physiological concentrations of CGRP inhibited pre-B cell responses to interleukin-7 (IL-7). Inhibition was specific in that it was blocked by the CGRP antagonist CGRP8-37. Adrenomedulin, substance P, and calcitonin had no effect on B cell precursor responses. Similar responses were observed with B220+/IgM- B cell precursors. Inhibition of IL-7 responses in B220+/IgM- cells suggests that CGRP has a direct effect on B cell precursors. Studies with cultured bone marrow adherent cells found that CGRP also has an indirect effect on IL-7 responses. Cultured bone marrow-adherent cells were treated with CGRP for 24 h, and anti CGRP was added to the supernatants to neutralize CGRP. Concentrations of CGRP as low as 0.01 nM induced a factor that inhibited colony formation. In contrast, CGRP did not induce an inhibitory factor in cultured bone marrow macrophages, suggesting that CGRP induces an inhibitory factor in some adherent cell other than macrophages. The results show that CGRP has both direct and indirect effects on developing B cells and support a role for CGRP as an inhibitor of early B cell development. PMID- 10811008 TI - Analysis of interferon-gamma-dependent and -independent pathways of macrophage activation. AB - Macrophages are a cellular cornerstone of the innate immune response. The outcome of macrophage activity during development of an immune response to microbes results from macrophage activation by both organism-derived and host-derived factors. In order to more fully understand the spectrum of responses expressed by macrophages when encountering these distinct stimuli, we investigated the similarities and differences between interferon-gamma receptor (IFN-gammaR) dependent macrophage activation and stimulation of macrophages through the Type A1 scavenger receptor (SR). We observed distinct patterns of macrophage activation depending on the nature of the ligand. IFN-gamma and the SR ligand lipotechoic acid (LTA) induced largely non-overlapping sets of genes. The use of two additional SR ligands, maleylated bovine serum albumin and the polydeoxynucleotide poly dI:dC, revealed differences within SR activation-induced gene expression. We also observed that priming with IFN-gamma resulted in an enhanced response to subsequent SR-mediated activation. These results suggest that full potentiation of macrophage activity during development of an antimicrobial immune response is achieved by activation of these cells through multiple receptors. PMID- 10811009 TI - Scavenger receptor-mediated delivery of muramyl dipeptide activates antitumor efficacy of macrophages by enhanced secretion of tumor-suppressive cytokines. AB - We showed that muramyl dipeptide (MDP) conjugated to maleylated bovine serum albumin (MBSA) was internalized by macrophages (Mphi) through scavenger receptor (SCR)-mediated endocytosis, which leads to 50-fold higher cytotoxic activity against non-Mphi tumor cells compared with that elicited by free MDP-treated Mphi. The enhanced cytotoxic effect of MBSA-MDP was found to be a result of higher secretion of interleukin (IL)-1, IL-6, tumor necrosis factor alpha (TNF alpha), and nitric oxide (NO) because the addition of antibodies directed against IL-1, IL-6, or TNF-alpha in combination with Mphi cultures totally abrogated the tumoricidal activity of MBSA-MDP. It is interesting to note that MBSA-MDP triggers the secretion of IL-12, whereas IL-10, a Mphi suppressor cytokine, could be detected only on free MDP treatment. The cytotoxic activity of MBSA-MDP was inhibited by indomethacin, indicating a regulatory role for prostaglandin E2 (PGE2). Efficient SCR-mediated intracellular delivery of MDP leading to elimination of cancer cells suggests the immunotherapeutic potential of this approach for treatment of neoplasia. PMID- 10811010 TI - Shared features of transcription: mutational analysis of the eosinophil/basophil Charcot-Leyden crystal protein gene promoter. AB - The lineage-specific Charcot-Leyden crystal (CLC) protein is found in human eosinophils and basophils where it comprises 7-10% of the cellular protein content. Previous work from our laboratory has identified the motif GGAGA[A/G] as a powerful enhancer of gene transcription ill two eosinophil ribonuclease genes. To evaluate a potentially larger role for this motif in the transcriptional regulation of eosinophil genes, we have isolated 1504 nucleotides 5' to the transcriptional start site of the gene encoding CLC protein and identified a functionally active promoter that includes three distinct copies of the GGAGAA motif. Destruction of only one of the three motifs by site-directed mutagenesis resulted in loss of promoter activity (73 +/- 6% reduction), suggesting that this core motif is necessary but not sufficient to support enhanced transcriptional activity. Sequence comparisons and site-specific mutagenesis has permitted further delineation of this enhancer element which, as a result of this work, is now defined as GGAGA[A/G]NNNA. Electromobility shift assays demonstrated specific binding of nuclear protein(s) from an eosinophilic clone-15 nuclear extract to this extended motif. Similar analysis of a GATA-1 binding site demonstrated enhancer activity, with mutagenesis resulting in a 94 +/- 1.4% reduction in activity, whereas the AML1 site functioned as a gene silencer. PMID- 10811011 TI - Differential chemokine receptor expression and function in human monocyte subpopulations. AB - The subset of human blood monocytes expressing low levels of CD14 and high levels of CD16 (CD14+CD16+) exhibits features resembling mature tissue macrophages and can be expanded in inflammatory conditions. We analyzed expression of CC chemokine receptors (CCR) in CD14+CD16+ versus CD14++ monocytes, which may be crucial for specific trafficking. Multicolor flow cytometric analysis of whole peripheral blood revealed that, as opposed to CD14++ monocytes, the CD14+CD16+ subset lacked surface expression of monocyte chemotactic protein-1 (MCP-1) receptor CCR2, however, it showed significantly higher surface expression of the macrophage inflammatory protein 1alpha (MIP-1alpha)/RANTES receptor CCR5. This was paralleled by differences in mRNA expression in the subsets, as shown by reverse transcriptase-polymerase chain reaction using sorted cells. In comparison to CD14++ monocytes, CD14+CD16+ cells expressed lower CCR2 but higher CCR5 transcript levels, whereas CCR1 levels were equivalent. Flow cytometric analysis of isolated human monocytes recovered after transendothelial chemotaxis assays revealed that the percentage of CD14+CD16+ cells was dramatically reduced in the fraction migrating toward MCP-1 compared with the fraction that did not migrate or the input, showing that polarized CCR2 expression was accompanied by a differential chemotactic responsiveness. Moreover, CD11b surface expression was preferentially up-regulated by MCP-1 in CD14++ cells but by MIP-1alpha in CD14+CD16+ monocytes, confirming the functional relevance of distinct CCR expression. The characteristics of CD14+CD16+ cells may reflect preactivation by cytokines and determine their predilective localization during specific inflammatory conditions or susceptibility to infection. PMID- 10811012 TI - Coordinate activation of endogenous p38alpha, beta, gamma, and delta by inflammatory stimuli. AB - The p38 family of mitogen-activated protein kinases is believed to mediate a variety of leukocyte responses to pro-inflammatory stimuli. There are four members of the p38 family, and although activation of the different members has been studied in transiently transfected cells much less is known about activation of the endogenous p38s, particularly in myeloid lineage cells. To investigate activation of endogenous p38s, we have made monoclonal antibodies specific for each p38 and have used these antibodies to study p38 activation by pro inflammatory stimuli in several human monocytic cell lines. Without stimulation endogenous p38alpha kinase activity was readily detectable, whereas that of p38beta, gamma, and delta was barely measurable. In response to inflammatory stimuli, we observed a time- and dose-dependent activation of all four p38s. The kinetics of activation of each of the p38s were similar for each stimulus used, suggesting a common upstream activation pathway. Simultaneous activation of the p38s suggests that all four may be important in inflammation. PMID- 10811013 TI - Expression of genes involved in initiation, regulation, and execution of apoptosis in human neutrophils and during neutrophil differentiation of HL-60 cells. AB - Neutrophils possess a very short lifespan, dying by apoptosis. HL-60 cells undergo apoptosis after neutrophil differentiation with dimethyl sulfoxide (DMSO). We have found that the onset of apoptosis in neutrophil-differentiating HL-60 cells correlates with the achievement of an apoptosis-related gene expression pattern similar to that of peripheral blood mature neutrophils. Using reverse transcriptase-polymerase chain reaction, cloning, and sequencing techniques, we have found that HL-60 cells express bak, bik, bax, bad, bcl-2, bcl xL, bcl-w, bfl-1, fas, and caspases 1-4 and 7-10. After DMSO treatment, bak, bcl w, bfl-1, fas, and caspases 1 and 9 were up-regulated, whereas bik, bcl-2, and caspases 2, 3, and 10 were down-regulated at different degrees, achieving mRNA expression levels that correlated with those detected in peripheral blood neutrophils. Caspase-2 mRNA and protein expression was drastically reduced after HL-60 cell differentiation, being absent in both HL-60-differentiated neutrophils and mature neutrophils, whereas caspase-3 and -10 mRNA and protein expression were diminished upon HL-60 cell differentiation until achieving the respective levels found in mature neutrophils. Bak and bfl-1 mRNA levels were largely increased during DMSO-induced differentiation of HL-60 cells, and these genes were the bcl-2 family members that were expressed most abundantly in mature neutrophils. Bcl-2 overexpression or caspase inhibition prevented differentiation induced apoptosis in HL-60 cells, but not their differentiation capability. Neutrophil spontaneous apoptosis was also blocked by the caspase inhibitor z-Asp 2,6-dichlorobenzoyloxymethylketone. Peripheral blood neutrophils expressed bak, bad, bcl-w, bfl-1, fas, and caspases 1, 3, 4, and 7-10, but hardly expressed bcl 2, bcl-xL, bik, bax, and caspase-2. These results suggest that the above gene expression changes in neutrophil-differentiating HL-60 cells may play a role in the acquisition of the neutrophil apoptotic features. PMID- 10811014 TI - A molecular analysis of NKT cells: identification of a class-I restricted T cell associated molecule (CRTAM). AB - cDNA library subtraction techniques were used to identify transcripts expressed by activated mouse alphabetaTCR+ CD4-CD8- (double-negative; DN) T cells, a subset of natural killer T (NKT) cells. The most frequent cDNAs identified included the chemokines TCA3, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and lymphotactin (LPTN), the cytokines interleukin-4 (IL-4) and interferon-gamma (IFN-gamma), and a granzyme. We also identified a new member of the immunoglobulin superfamily (Ig-SF). This molecule was designated class I restricted T cell-associated molecule (CRTAM) as a result of its restricted expression pattern in T cells. Human CRTAM was also identified, and shares the same expression pattern as the mouse molecule. LPTN and CRTAM exhibit the same expression pattern in T cells, suggesting the existence of a gene expression program common to class I-MHC-restricted T cells. PMID- 10811015 TI - Ceramide-mediated stimulation of inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF) accumulation in murine macrophages requires tyrosine kinase activity. AB - In macrophages, bacterial lipopolysaccharide (LPS) has been noted to mimic certain effects of the sphingolipid ceramide, suggesting that ceramide may be involved in macrophage activation by LPS and/or that LPS utilizes ceramide related signaling pathways. Putative downstream targets of ceramide include a ceramide-activated (serine/threonine) protein kinase (CAPK) and phosphatase (CAPP). However, the potential role of tyrosine phosphorylation pathways in macrophage response to ceramide has not been examined. Herein we report that cell permeable analogs of ceramide up-regulate both inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF) production in RAW 264.7 murine macrophages. Herbimycin A and genistein, potent natural inhibitors of protein tyrosine (but not serine/threonine) phosphorylation, block ceramide-induced iNOS and TNF production. Furthermore, the highly src-family selective pyrazolopyrimidine inhibitor PP1 also blocks ceramide-induced iNOS and TNF production in RAW 264.7 cells. We found that PP1 also inhibits ceramide-mediated tyrosine phosphorylation of the src-family kinase hck. These data indicate that src-related tyrosine kinases play a critical role in macrophage activation by ceramide. PMID- 10811016 TI - Positive regulation of human T cell activation by Gi2 proteins and interleukin-8. AB - We investigated whether pertussis toxin (PT)-sensitive heterotrimeric Gi proteins (Gi1, Gi2, Gi3) are involved in the regulation of TCR-induced activation of human T cells. First, Gi proteins were inactivated by PT: pretreatment with PT of purified blood T lymphocytes before CD3 cross-linking inhibited cell proliferation (-71.1 +/- 22.0%, P < 0.001), production of interleukin-2 (IL-2; 47.3 +/- 12.6%, P = 0.008), and expression of CD25 (-24.6 +/- 11.7%, P < 0.001) and CD69 (-25.7 +/- 9.0%, P < 0.001). Then, to identify which of the three Gi was involved, Gi1, Gi2, and Gi3 proteins were specifically inactivated by stably transfecting dominant-negative mutated forms of their alpha subunit in Jurkat cells. After activation, IL-2 production and CD69 expression were inhibited only in cells expressing inactive Gi2. We then studied the effects of interleukin-8 (IL-8), a CXC-chemokine with receptors coupled to Gi2 and produced in an autocrine fashion by activated T cells. Although its effects varied among donors, exogenous IL-8 stimulated proliferation and CD25 expression (up to, respectively, 200 and 77%) of PB T lymphocytes in response to CD3 activation, in a PT-sensitive manner. IL-8 also stimulated IL-2 production (by up to 42%) and CD69 expression, although weakly (+27%). Anti-human IL-8 antibody inhibited proliferation (-43%) and CD25 up-regulation (-45%) of activated T lymphocytes. In summary, several major responses of human T lymphocytes to TCR-mediated activation are regulated by Gi2 proteins, which for this function can be activated by IL-8 in an autocrine manner. PMID- 10811017 TI - Abnormal down-regulation of PKC is responsible for giant granule formation in fibroblasts from CHS (beige) mice--a thiol proteinase inhibitor, E-64-d, prevents giant granule formation in beige fibroblasts. AB - We have previously reported that the abnormally rapid down-regulation of protein kinase C (PKC) activity is responsible for the cellular dysfunction in natural killer (NK) cells and polymorphonuclear leukocytes (PMNs) from Chediak-Higashi syndrome (beige) mice. In this report, we examined whether the down-regulation of PKC is associated with giant granule formation in fibroblasts from beige mice. In cultured beige fibroblasts, the membrane-bound PKC activity declined significantly after phorbol ester stimulation. We found that E-64-d, which is a thiol proteinase inhibitor and protects PKC from calpain-mediated proteolysis, reversed the declined PKC activity and prevented giant granule formation in beige fibroblasts. Moreover, E-64-d corrected the reduced lysosomal elastase and cathepsin G activity in beige fibroblasts. In contrast, specific PKC inhibitors, chelerythrin and calphostin C, promoted giant granule formation in normal fibroblasts. We also demonstrate that ceramide production is enhanced in beige fibroblasts and is involved in the rapid down-regulation of PKC. These results suggest that the accelerated breakdown of PKC observed in beige fibroblasts is caused by enhanced ceramide production and is also responsible for giant granule formation. PMID- 10811018 TI - Phospholipase A2 and its potential regulation of islet function. PMID- 10811019 TI - Assessment of pancreatic enzyme secretory capacity by a modified Lundh test. AB - BACKGROUND: The Lundh test is a usual means of estimating the enzyme secretory capacity of the gland. During this procedure, however, a major proportion of the test meal is removed from the duodenum together with the gastric, duodenal, and pancreatic secretions and the bile. This study was undertaken to compare the pancreatic enzyme secretion induced by the Lundh procedure with that resulting from stimulation of the normal digestive process, by reinfusion of the aspirated duodenal juice. METHODS: Nine men (mean age: 46.7, range 42-55 yr) free from pancreatic disease were studied. Pancreatic secretion was measured via a multiple lumen tube by aspiration of the duodenal juice. After a basal period the Lundh test meal was placed in the stomach and the duodenal juice was completely aspirated. On a separate day, the procedure was repeated, but the aspirated duodenal juice was reinfused into the upper jejunum. RESULTS: In the first 30 min of the test period, the enzyme outputs were the same on both test days. In the 30 60-min period, the lipase output, and in the 75-90-min period, the amylase output was significantly lower during the Lundh test compared with the jejunal reinfusion test. The CCK levels were significantly above the basal level at 20 and 40 min, but the increase was significantly lower during the traditional Lundh test. No significant difference in gastrin release was observed during either the Lundh or the reinfusion test. CONCLUSIONS: In the traditional Lundh test, the trypsin secretory capacities of the gland are measured appropriately, but the lipase and amylase secretory capacity and the CCK release are not fully represented compared with the reinfusion test. An association between the lower CCK release and lipase amylase secretion is suggested. PMID- 10811020 TI - Bone mineral metabolism, bone mineral density, and body composition in patients with chronic pancreatitis and pancreatic exocrine insufficiency. AB - BACKGROUND: Calcium and vitamin D homeostasis seem to be abnormal in patients with exocrine pancreatic dysfunction resulting from cystic fibrosis. Only a few studies have evaluated and described bone mineral metabolism in patients with chronic pancreatitis and pancreatic insufficiency. METHODS: Thirty-two patients with chronic pancreatitis and residual exocrine pancreatic function (group 1) and 26 patients with pancreatic exocrine insufficiency (i.e., meal-stimulated intraduodenal lipase <10% of lowest normal range and steatorrhea) (group 2) were studied. Serum levels of total calcium, phosphate, 25 (OH)D, 1.25(OH)2D, alkaline phosphatase, and parathyroid hormone were measured. Bone mineral density (BMD), bone mineral content (BMC), lean body mass (LBM), and fat mass (FM) were measured using a dual-energy X-ray absorptiometry (DXA) scanner. RESULTS: Alcohol was a causative factor in 79% of the patients. Fifty-six percent in group 1 and 69% in group 2 had Z-scores of the BMD < -1. The mean Z-score was -1.16 +/- 1.29 in group 1 and -1.32 +/- 0.90 in group 2. The mean Z-score of the BMC was -1.02 +/- 1.17 vs -1.39 +/- 0.987. In both groups mean 25 (OH)D and mean 1.25(OH)2D were below reference range. Plasma concentrations of albumin-corrected calcium, alkaline phosphatase, and parathyroid hormone were in the upper range of the reference range. Mean Z-scores of LBM were -0.69 +/- 1.34 in group 1 vs -1.01 +/- 1.12 in group 2 and Z-scores of FM were -0.27 +/- 1.70 in group 1 vs -0.95 +/- 1.01 in group 2 (p <0.05). CONCLUSION: Patients with chronic pancreatitis, in particular patients with advanced disease and steatorrhea, are at risk of developing significant bone loss. Despite normal body mass index the patients are characterized by loss of lean body mass and fat mass. The present study shows that these patients have decreased serum levels of vitamin D metabolites and low bone mass. PMID- 10811022 TI - Inositol 1,4,5-trisphosphate formation, cytoplasmic calcium dynamics, and alpha amylase secretion of pancreatic acini isolated from aged and chronically alcohol fed rats. AB - METHODS: Three-month-old female Wistar rats were fed with 20% alcohol in their drinking fluid over 6-17 mo using an interrupted feeding regimen. At different times, pancreatic acini were isolated by mild collagenase digestion. The concentrations of inositol-1,4,5-trisphosphate (1,4,5-IP3) were determined by a specific radioreceptor assay, before and at different times after stimulation with varying concentrations of CCK-8. CCK-induced dynamics of cytoplasmic calcium ([Ca2+]c) was investigated in acinar cells by confocal laser raster microscopy. Acinar alpha-amylase (Aml) secretion was measured as enzyme activity in the medium compared to the total activity in the suspension. RESULTS: In 12-13-mo-old rats, the CCK-stimulated 1,4,5-IP3 formation in acini was found to be decreased compared to young rats (age 4 mo). In rats of the same age fed with ethanol from the age of 3 mo on, 1,4,5-IP3 concentrations in acini were higher and reached values comparable to those in young rats. Correspondingly, the CCK-induced [Ca2+]c dynamics in acini isolated from 9-mo-old rats was impaired compared to that of young rats but normal in aged, chronically alcohol-fed rats. Aml secretion under CCK stimulation, however, which was decreased in aged rats, was additionally impaired after alcohol feeding. CONCLUSION: Chronic alcohol feeding modifies 1,4,5-IP3 formation, the [Ca2+]c dynamics of, and the Aml secretion of rat pancreatic acini in response to CCK stimulation. Obviously, the age-related impairment of 1,4,5-IP3 formation and [Ca2+]c dynamics is improved. In contrast, the decrease in Aml secretion of acini isolated from aged rats is more pronounced after long-term alcohol-feeding. PMID- 10811021 TI - Role of various phospholipases A2 and inhibitors in the pathogenesis and prevention of pancreatic acinar cell necrosis: studies with isolated rat pancreatic acini. AB - BACKGROUND: Phospholipase A2 (PLA2) may play a central role in the pathogenesis of pancreatic acinar cell necrosis. Several questions, however, are unsolved: Is acinar cell necrosis caused by PLA2 derived from infiltrating leukocytes or from pancreatic PLA2 itself? Does PLA2 cause cellular lysis by the release of lysolecithin from lecithin or by generation of free radicals? The aims of this study were to determine which form of PLA2 is responsible for cellular damage and how to inhibit its action. METHODS: Isolated rat pancreatic acini were prepared by collagenase digestion. Newly synthesized proteins were labeled by 35S methionine. Acini were incubated in buffer to which various factors, such as porcine pancreatic PLA2 or bee venom PLA2, homogenates of either leukocytes or pancreatic homogenates, all with or without lecithin and with or without potential inhibitors (aprotinin, 4-bromophenacylbromide, BM 16.2115, quinacrine, various analogs of arachidonic acid), or free radicals (hydrogen peroxide, xanthine/ xanthine oxidase) with or without allo-purinol or dismutase/catalase were added. Cellular destruction was measured by the release of radiolabeled proteins. RESULTS: PLA2 alone, free radicals, and granulocytes were not harmful to acini within 30 min of incubation. Free radicals caused significant release of radiolabeled proteins only after 3 h of incubation; this release could be inhibited by scavengers. Incubation of pancreatic acini with PLA2 in combination with lecithin caused rapid release of radiolabeled proteins. Addition of high concentrations of enterokinase activated pancreatic homogenates both alone and with lecithin caused release of cellular proteins, suggesting that pancreatic PLA2 uses lecithin from pancreatic membranes as substrate. Almost all tested potential inhibitors of PLA2 were unable to prevent the destruction caused by either pancreatic or bee venom PLA2 and lecithin. However, HK 42, a polyunsaturated fatty acid analog, was able to reduce dose dependently the release of acinar proteins caused by pancreatic PLA2 and lecithin. CONCLUSION: Pancreatic PLA2 and not PLA2 from infiltrating leukocytes may play a role in pancreatic acinar cell necrosis. Cellular lysis is caused upon the action of lysolecithin and probably not via the action of free radicals. PMID- 10811023 TI - Biliary and gastric bypass or stenting in nonresectable periampullary cancer: analysis on the basis of controlled trials. AB - BACKGROUND: The median survival rate of patients with nonresectable periampullary cancer is not much longer than 6-12 mo. Nevertheless, in most incurable patients palliative treatment is necessary, which has to focus on jaundice, pain, and prevention of gastric outlet obstruction. Up to now, debate remains about how to best provide palliative treatment. METHOD: The results of controlled clinical trials and large multicenter studies comparing operative biliary bypass and biliary stent insertion in nonresectable pancreatic tumors are discussed in this review. RESULTS: The initial success rate in palliation of jaundice is similar after endoscopic stent insertion and biliary bypass operation (range: 90-95 %). Morbidity (range: 1 1-36% vs 26-40%) and 30-d mortality (range: 8-20% vs 15-31%) is higher after bypass operation, whereas stent insertion is accompanied by a higher rate of hospital readmission and reintervention because of recurrent jaundice (range: 28-43%) and a later gastric outlet obstruction (up to 17%). CONCLUSION: Endoscopic biliary stent insertion should be performed if there is evidence of hepatic, peritoneal, or pulmonary metastasis formation, in old patients with a high comorbidity, or if the patient has had several laparotomies. Combined biliary and gastric operative bypass procedures should be performed in nonresectable periampullary carcinomas with accompanying gastric outlet obstruction, in the absence of metastatic spread, if a locally advanced tumor is the only reason for incurability, if exploratory laparotomy demonstrates an unresectable tumor, or if endoscopic treatment fails. PMID- 10811024 TI - Inhibitory action of the anomers of 2-deoxy-D-glucose tetraacetate on the metabolism of D-glucose in rat pancreatic islets. AB - BACKGROUND: The tetra-acetate ester of 2-deoxy-D-glucose was recently found to either inhibit or augment insulin secretion, depending on the concentration of the ester. Both the positive and negative insulinotropic actions of the ester display anomeric specificity. METHODS: The effects of the alpha- and beta-anomer of 2-deoxy-D-glucose tetra-acetate (5.0 mM) on the metabolism of D-[5-3H]glucose and D-[U-14C]glucose (8.3 mM) were investigated in isolated rat pancreatic islets. RESULTS: Both the alpha- and beta-anomers of 2-deoxy-D-glucose tetra acetate inhibited the generation of 3HOH from D-[5-3H]glucose and that of 14CO2, as well as radioactive acidic metabolites and amino acids, from D-[U-14C]glucose. They also lowered the paired ratio between D-[U-14C]glucose oxidation and D-[5 3H]glucose utilization. No significant anomeric difference could be detected, however, in these experiments. CONCLUSIONS: The effects of the alpha- and beta anomer of 2-deoxy-D-glucose tetra-acetate on the metabolism of D-glucose in isolated rat pancreatic islets reinforce the view that the insulinotropic action of monosaccharide esters involves a dual mode of action, linked to both the metabolic effects of their glucidic moiety and a direct interaction of the esters themselves with a stereospecific receptor system. PMID- 10811025 TI - A case of pancreatic carcinoma with marked ductal dilatation: what contributed to the dilatation? AB - BACKGROUND: We report the case of an 82-yr-old man with invasive ductal carcinoma of the pancreatic head, in which the main pancreatic duct and duct of Santorini were markedly dilated, measuring 1.6 and 1.1 cm, respectively, in diameter on computed tomography. METHODS: A preoperative diagnosis of ductal carcinoma of the pancreatic head was made, and Whipple's procedure was carried out. RESULTS: Histopathologically, the tumor was diagnosed as moderately differentiated tubular adenocarcinoma, and the resected pancreatic parenchyma showed low papillary mucous cell hyperplasia and atypical hyperplasia in dilated ductular branches. Conclusion. Even among patients with tubular adenocarcinoma, the most common type of pancreatic ductal carcinoma, if the patient is aged and has chronic pancreatitis, the main pancreatic duct and duct of Santorini may dilate to the same degree as in mucin-hypersecreting neoplasm. PMID- 10811026 TI - Solid and cystic tumor of the pancreas: clinicopathologic and genetic studies of four cases. AB - BACKGROUND: Solid and cystic tumor (SCT) of the pancreas can be distinguished from other pancreatic neoplasms by its nearly exclusive occurrence in young women, and its favorable prognosis after complete resection. METHODS: We experienced four cases with SCT of the pancreas, and analyzed these tumors by immunohistochemical and electron microscopic studies, as well as genetic analysis of ras oncogene mutation. RESULTS: The presented cases expressed the neuron specific enolase in two cases, alpha1-antitrypsin and alpha1-antichymotrypsin in two cases, and vimentin in one case, which indicated that this tumor originates from pleuripotential embryonic stem cells. No patients had mutations of K-ras gene in codon 12, and further genetic analysis is required to predict the malignant potential. CONCLUSION: SCT of the pancreas appears to have limited malignant potential and the metastatic ratio is not high, although the tumor has local invasion. Therefore, an aggressive surgical approach seems fully justified. PMID- 10811027 TI - Solid pseudopapillary tumor of the pancreas. AB - BACKGROUND: Its considerable size at the time of diagnosis and low grade of malignancy are typical features of the solid-pseudopapillary tumor, which has a tendency to predominantly affect young females. A relationship to the long-term intake of oral contraceptives is discussed. Invasive tumor growth or metastases have been observed only rarely until now. METHODS: The 53-yr-old female patient we report on here was treated by radical partial pancreatoduodenectomy for a nonmetastasizing solid-pseudopapillary tumor of the pancreatic head (T1bN0M0) 19 mo ago. RESULTS: Histopathological studies made a definitive diagnosis of solid pseudopapillary tumor. The patient is recurrence-free, and there are no signs of metastases at present. Since a microscopically invasive tumor growth is assumed, oncologically curative resection should be preferred vs the less radical enucleation. CONCLUSION: In this report, a case of the rare solid-pseudopapillary tumor of the pancreas is described. In contrast to other pancreatic tumors, the semimalignant solid-pseudopapillary tumor has a favorable prognosis. PMID- 10811028 TI - Treatment of chyloperitoneum after extended lymphatic dissection during duodenopancreatectomy. AB - BACKGROUND: Chyloperitoneum is a rare postoperative complication that might be caused by an interruption of chylous ducts in the mesenteric root or the cysterna chyli. Two cases of chyloperitoneum after duodenopancreatectomy are reported in the literature. METHODS: We here report the third case that developed a chyloperitoneum 2 wk postoperatively when he resumed his normal diet. RESULTS: The patient was treated conservatively with paracenteses and chyloperitoneum subsided thereafter. CONCLUSIONS: Chyloperitoneum after extended duodenopancreatectomy might be treated conservatively. PMID- 10811029 TI - Alkaloidal sugar mimetics: biological activities and therapeutic applications. AB - Alkaloids mimicking the structures of sugars inhibit glycosidases because of a structural resemblance to the sugar moiety of the natural substrate. Glycosidases are involved in a wide range of important biological processes, such as intestinal digestion, post-translational processing of glycoproteins and the lysosomal catabolism of glycoconjugates. The realization that alkaloidal sugar mimics might have enormous therapeutic potential in many diseases such as viral infection, cancer and diabetes led to increasing interest and demand for these compounds. In this review, the structural basis of the specificity of alkaloidal sugar mimics and their current and potential applications to biomedical problems are reviewed. PMID- 10811030 TI - Protease inhibitors. Part 2. Weakly basic thrombin inhibitors incorporating sulfonyl-aminoguanidine moieties as S1 anchoring groups: synthesis and structure activity correlations. AB - Two series of derivatives have been prepared and assayed as inhibitors of two physiologically relevant serine proteases, human thrombin and human trypsin. The first series includes alkyl-/ aralkyl-/aryl- and hetarylsulfonyl-aminoguanidines. It was thus observed that sulfanilyl-aminoguanidine possesses moderate but intrinsically selective thrombin inhibitory properties, with KI values around 90 and 1400 nM against thrombin and trypsin respectively. Further elaboration of this molecule afforded compounds that inhibited thrombin with KI values in the range 10-50 nM, whereas affinity for trypsin remained relatively low. Such compounds were obtained either by attaching benzyloxycarbonyl- or 4 toluenesulfonylureido-protected amino acids (such as D-Phe, L-Pro) or dipeptides (such as Phe-Pro, Gly His, beta-Ala-His or Pro-Gly) to the N-4 atom of the lead molecule, sulfanilyl-aminoguanidine, or by attaching substituted-pyridinium propylcarboxamido moieties to this lead. Thus, this study brings novel insights regarding a novel non-basic S1 anchoring moiety (i.e., SO2NHNHC(=NH)NH2), and new types of peptidomimetic scaffolds obtained by incorporating tosylureido-amino acids/pyridinium-substituted-GABA moieties in the hydrophobic binding site(s). Structure-activity correlations of the new serine protease inhibitors are also discussed based on a QSAR model described previously for a large series of structurally-related derivatives (Supuran et al. (1999) J. Med. Chem., in press). PMID- 10811032 TI - Influence of stereoisomers of 4-fluoroglutamate on rat brain glutamate decarboxylase. AB - Inhibition of rat brain glutamate decarboxylase (GAD, EC 4.1.1.15) by individual stereoisomers of 4-fluoroglutamate (4-F-Glu) and 2-fluoro-4-aminobutyrate (2-F GABA) was studied. All stereoisomers of 4-F-Glu inhibited decarboxylation of L glutamate catalysed by the enzyme preparation. At 1 x 10(-2) M concentration, the most potent inhibitor of GAD was D-erythro-4-F-Glu with about 70% inhibition in the presence of 1.23 x 10(-2)M L-glutamate. The inhibition by all stereoisomers was of the competitive type. Ki values ranged from 2 x 10(-3)M for the D-erythro isomer to 1.1 x 10(-2)M for the D-threo and L-erythro isomers. The influence of all stereoisomers was reversible as shown by dialysis except for a small amount in the case of the D-erythro isomer. The inhibition was independent of external pyridoxal-5'-phosphate added. No inhibition of rat brain GAD was found with 2 fluoro-4-aminobutyrate stereoisomers. PMID- 10811031 TI - Use of a bisubstrate inhibitor to distinguish between isocitrate dehydrogenase isozymes. AB - Bisubstrate inhibitors, obtained by covalently linking 2-oxoglutarate with NAD+ and NADP+, were synthesized and tested for their ability to inhibit NAD+- and NADP+-dependent isocitrate dehydrogenases from pig heart mitochondria. The NADP+ dependent enzyme was specifically inhibited by the NADP oxoglutarate adduct and not by the NAD adduct. The NADP adduct was competitive with both coenzyme and substrate, isocitrate. In contrast, the NAD+-dependent enzyme was inhibited by both adducts. NAD oxoglutarate is competitive with both NAD+ and isocitrate while the NADP adduct is competitive with isocitrate but not with NAD+. Nevertheless conditions could be set up so that use of these inhibitors would be feasible for a metabolic study. PMID- 10811033 TI - Inhibition of rabbit muscle isozymes by vitamin C. AB - The ability of vitamins C, E and K to inhibit enzymes directly has been investigated. It was found that vitamin E and some analogs and menadione (vitamin K3) inhibited several enzymes irreversibility at concentrations below one millimolar. Ascorbate inhibits rabbit muscle 6-phosphofructokinase (MPFK-1; EC 2.7.1.11), muscle type LDH (EC 1.1.1.27), and muscle AK (EC 2.7.4.3) at low concentrations that do not inhibit equivalent liver isozymes. Ascorbate Ki values for muscle-type LDH and heart-type LDH isozymes are 0.007 and 3 mM, respectively. The ascorbate Ki value for rabbit skeletal muscle PFK-1 is 0.16 mM; liver PFK-I is not inhibited by ascorbate. Dehydroascorbate does not inhibit any enzyme at ascorbate concentrations normally found in cells. All ascorbate inhibitions are completely reactivated or nearly so by L-ascorbate oxidase, CYS, GSH, or DTT. We propose a hypothesis that ascorbate facilitates glycogen storage in muscle by inhibiting glycolysis. The relationship between ascorbate metabolism and diabetes is discussed. PMID- 10811034 TI - Carbonic anhydrase inhibitors; phosphoryl-sulfonamides--a new class of high affinity inhibitors of isozymes I and II. AB - A series of phosphorylated aromatic/heterocyclic sulfonamides with the general formula ArSO2NHPO3H2 have been prepared by condensing ArSO2NH2 with phosphorus pentachloride, followed by controlled hydrolysis in the presence of formic acid. The new derivatives generally act as stronger inhibitors of two carbonic anhydrase (CA) isozymes, CA I and CA II, as compared to the parent unsubstituted sulfonamides from which they were obtained. The inhibition mechanism by this new class of CA inhibitors, as well as structure activity correlations for the series of investigated derivatives, are also discussed. PMID- 10811035 TI - The doctor, the patient and the body. AB - Michael Balint has helped us realize that the clinical interaction is, above all, an inter-human interaction. However, the lived experience of the body -- the existential anatomy -- occupies but a very small space in the consciousness of medicine. Even the "psychosomatic body" is a theoretical construct favouring observation rather than understanding. The doctor shares the conditions of embodied life with his patients, and, in addition, has the opportunity to refine his sensitivity to the variations of bodily experience in his work with patients. Thus, a bodily empathic basis for clinical interaction is laid on which understanding and diagnosis may reinforce one another in a common endeavour. PMID- 10811036 TI - Primary care physicians and their information-seeking behaviour. AB - OBJECTIVE: To investigate primary care physicians' continuing medical education (CME) and their information-seeking behaviour and to compare it with that of hospital doctors DESIGN: Postal survey of Norwegian physicians. SUBJECTS: 1276 physicians (response rate 78%), 283 primary care physicians, 741 hospital doctors. MAIN OUTCOME MEASURES: Self-perceived ability to cope with medical knowledge and self-reported CME activities. RESULTS: Two out of three doctors thought they could cope with the increasing flow of medical information. Courses, meetings and congresses were considered the most important CME activities. Primary care physicians spent less than 3 hours per week on medical reading, compared with more than 4.5 hours among hospital doctors; 59% of primary care physicians had access to the Internet compared with 76% among hospital doctors. Time spent on medical reading and formalized courses decreased from 1993 to 1999 for all groups of physicians. CONCLUSION: Primary care physicians rely on several information sources in their professional updating. They pay less attention to informal ways of learning than their hospital colleagues do. PMID- 10811037 TI - Sense of coherence in different stages of health and disease in northern Sweden- gender and psychosocial differences. AB - OBJECTIVE: To investigate "Sense of Coherence" (SOC) and its relation to perceived health, different stages of disease, and different psychosocial factors in a population-based study. DESIGN: Postal survey of a population-based sample, the MONICA study (1994). SETTING: Norrbotten and Vasterbotten, the two northernmost counties in Sweden, with a total population of 510000 inhabitants. SUBJECTS: 837 men and 882 women in three mutually-exclusive groups: stomach trouble of many years' standing, identified disease (stroke, cardiac infarction, diabetes, anti-hypertension treatment) and no reported disease. MAIN OUTCOME MEASURES: SOC scores in relation to sociodemographic variables and perceived health. RESULTS: We found a relationship between low SOC scores and poor perceived health, low social support and low emotional support on a population level. When comparing persons with stomach trouble with those without disease, or with established diseases, we found similar relationships between low mean SOC scores in all strata for both women and men. "Perceived health", however, was only significantly correlated for women, and women had an overall stronger relationship. CONCLUSIONS: In a study in northern Sweden, female patients with stomach trouble comprise a vulnerable group. The concept of SOC introduces a new dimension for perceiving health and disease. In clinical practice, care providers can identify and elaborate on the relationship between SOC scores and sociodemographic data. PMID- 10811038 TI - List patient system: straitjacket or a tool for developing general practice? General practitioners' experiences from a pilot project in Norway. AB - OBJECTIVE: To investigate attitudes and experiences among general practitioners (GPs) taking part in an experiment with a list patient system in four municipalities in Norway. DESIGN: A questionnaire distributed to all 160 GPs in the four municipalities after 3 years' trial of the system. SETTING: Primary health care in Norway. MAIN OUTCOME MEASURES: The opinions of physicians on a list patient system after 3 years of trial. RESULTS: Most of the physicians held that the list patient system imposed more obligations than they were used to. However, the majority also held that the system enhanced preconditions for doing a good professional job; 68% wanted the system on a permanent basis, while 14% rejected it. CONCLUSION: Positive aspects outweigh negative aspects in the attitude of GPs to a list patient system. PMID- 10811039 TI - Predictive value of parent-reported symptoms in the assessment of otitis media with effusion during infancy. AB - OBJECTIVE: To determine the predictive value of parent-reported symptoms in the assessment of otitis media with effusion (OME) during infancy. DESIGN: Prospective longitudinal study that compared the results of questionnaire-based parental reports with the state of the middle ear assessed by otoscopy and tympanometry at 3-month intervals from birth to the age of 2 years. SETTING: Outpatient Department, University Hospital Maastricht, The Netherlands. SUBJECTS: One hundred and fifth healthy-born infants. MAIN OUTCOME MEASURES: Sensitivity, specificity, positive and negative predictive values of parent-reported ear infection, hearing loss, mouth breathing, snoring and common cold. RESULTS: Prevalence rates showed the following ranges: OME, 39%-53%, parent-reported ear infection, 2%-20%, hearing loss 2%-7%, mouth breathing, 30%-41%, snoring, 31%-41% and common cold, 65%-81%. Common cold demonstrated the highest sensitivity (83%) with a low specificity of 36%. Hearing loss gave the highest positive predictive value (PPV) (70%) with a moderate negative predictive value (NPV) (54%). PPV increased with the number of anamnese questions answered positively. CONCLUSIONS: The diagnostic value of parent-reported ear infection, hearing loss and common cold was found to be limited. Combinations of positively-reported symptoms gives increased diagnostic validity while the absence of parent-reported symptoms does not necessarily indicate the absence of OME. PMID- 10811040 TI - General practitioners' patterns of antimicrobial drugs prescription in the therapy of acute pharyngitis. AB - OBJECTIVE: To analyse the patterns of antibiotic prescription in the treatment of acute pharyngitis among a group of general practitioners (GPs). DESIGN: From the records of their patients, 11 specialists in general practice have collected post hoc data on their patients and on acute pharyngitis in the period from October 1, 1994 to September 30, 1995. SETTING: 11 teaching practices in four health centres in Zagreb, Croatia. SUBJECTS: GPs grouped according to whether they had pre school children in their care. MAIN OUTCOME MEASURES: The choice and cost of the prescribed antimicrobial drug, and the impact of the characteristics of the population in care to the prescription rate and structure. RESULTS: An antimicrobial drug was prescribed in 784 (72.1%) cases of pharyngitis. Two groups of GPs have shown significant differences in the prescription rate (p < 0.001), and in the choice of antimicrobial drugs (p < 0.001). CONCLUSION: GPs with a larger number of patients in their care suffering from chronic diseases tend to prescribe antimicrobial drugs more often in the treatment of acute pharyngitis. PMID- 10811041 TI - Urinary tract infections, antibiotic resistance and sales of antimicrobial drugs- an observational study of uncomplicated urinary tract infections in Icelandic women. AB - OBJECTIVES: To analyse the antimicrobial susceptibility pattern of bacteria causing symptomatic but otherwise uncomplicated lower urinary tract infections (UTI) in primary health care and the sales of antimicrobial drugs. SETTING: Primary health care in Akureyri District, Northern Iceland, with about 17400 inhabitants. PATIENTS: A total of 516 episodes of symptomatic but otherwise uncomplicated lower UTI in women 10 to 69 years of age. MAIN OUTCOME MEASURES: Number of verified UTI, bacterial species, antimicrobial susceptibility pattern, and total sales of antimicrobial drugs. RESULTS: Escherichia coli was by far the most common cause of UTI (83%), followed by Staphylococcus saprophyticus (7%). Infections caused by E. coli resistant to ampicillin accounted for 36% of cases, with the corresponding figures for sulfafurazol being 37%, cephalothin 45%, trimethoprim 13% and mecillinam 14%. Only 1% of the strains were resistant to nitrofurantoin. The total use of antimicrobial drugs was 17.4 DDD/1000 inhabitants/day. CONCLUSIONS: The resistance of bacteria causing uncomplicated UTI to common antimicrobials is high and must be taken into account when selecting treatment strategies. High consumption of antibiotics in the community indicates possible association. PMID- 10811042 TI - Randomised controlled trial of CRP rapid test as a guide to treatment of respiratory infections in general practice. AB - OBJECTIVE: To assess whether the frequency of antibiotic prescriptions to patients with respiratory infections is reduced when general practitioners (GPs) use a C-reactive protein (CRP) rapid test in support of their clinical assessment, and to study whether using the test will have any effect on the course of disease DESIGN: Randomised controlled trial. SETTING: 35 general practices, County of Funen, Denmark. PATIENTS: 812 patients with respiratory infection. MAIN OUTCOME MEASURES: Frequency of antibiotic prescriptions and morbidity 1 week after the consultation, as stated by the patients. RESULTS: In the CRP group the frequency of antibiotic prescriptions was 43% (179/414) compared with 46% (184/398) in the control group (odds ratio (OR) = 0.9, NS). After 1 week, increased or unchanged morbidity was stated more frequently in the CRP group (12%) than in the control group (8%) (OR = 1.6, p = 0.05). In the control group, the variable having the greatest influence on whether the GP prescribed antibiotics was the patients' general well-being (OR = 2.9, p < 0.0001), whereas in the CRP group the CRP value had the greatest influence (OR = 1.1 per unit increase (mg/l), p < 0.0001). CONCLUSION: Based on the present study, the use of the CRP rapid test in support of a possible antibiotic treatment for respiratory infections in general practice cannot be recommended. PMID- 10811043 TI - Randomised, double blind, placebo controlled trial of penicillin V in the treatment of acute maxillary sinusitis in adults in general practice. AB - OBJECTIVE: To compare the effectiveness of penicillin V with placebo in the treatment of adult patients with acute maxillary sinusitis in general practice. DESIGN: Randomised, double blind, placebo controlled trial. SETTING: 26 Danish general practices. PATIENTS: 133 adult patients with a clinical diagnosis of acute maxillary sinusitis based on maxillary pain and raised values of either C reactive protein (CRP) or erythrocyte sedimentation rate (ESR). MAIN OUTCOME MEASURES: Pain score and illness score as well as measurement of CRP and ESR values after initiation of treatment. RESULTS: Penicillin V led to a better recovery than did placebo. The difference was statistically significant 3 days after the initiation of treatment with regard to pain reduction, whereas no significant difference was found with regard to the reduction in the sense of illness. At the end of the study, significantly more patients in the penicillin group were completely free of pain compared to the placebo group. This difference was only found in patients with an initial pain score of more than three. The cure rate was 71% in the penicillin group and 37% in the placebo group. Significantly more patients achieved normal CRP values when treated with penicillin (88%) as opposed to placebo (75%). CONCLUSION: Penicillin V is more effective than placebo in the treatment of acute maxillary sinusitis in adults in general practice, but only in patients with pronounced pain. PMID- 10811044 TI - Variations in sick-listing practice among male and female physicians of different specialities based on case vignettes. AB - OBJECTIVE: To analyse the influence on sick-listing of speciality and sex of the prescribing doctor and of patients' way of presenting their problem. DESIGN: A sample of 360 general practitioners (GPs), 180 psychiatrists and 180 orthopaedic surgeons were presented case vignettes and asked to fill in a sick certificate for each case. The cases were chosen to reflect common causes of sick-listing in Sweden. The vignettes for each case were presented in three versions with uniform biomedical information plus a variation in the patient's attitude to sick listing. SETTING: Swedish general practice and hospital physicians. MAIN OUTCOME MEASURES: Proportion of cases not being sick-listed and costs for the period certified. RESULTS: The most important factor affecting sick-listing was the patient's attitude to sick-listing. Patients wishing sick-listing were sick listed to a greater extent than those who were reluctant. In addition, GPs sick listed more than orthopaedic surgeons and less than psychiatrists. Female doctors sick-listed more than male doctors, irrespective of speciality and patient attitude. CONCLUSION: Sick-listing is influenced by the physician's speciality and sex. Doctors are strongly influenced by how the patients present their problem. PMID- 10811045 TI - Cost-minimisation analysis of three conservative treatment programmes in 180 patients sick-listed for acute low-back pain. AB - OBJECTIVE: To perform a cost-minimization analysis of three conservative treatment regimes for acute low-back pain (LBP). DESIGN: A prospective randomized clinical trial. Patients were assigned at random to one of three treatment programmes: General Practitioner Programme (GPP-) controls, Manual Therapy Programme (MTP) or Intensive Training Programme (ITP). SETTING: Primary care and physiotherapists in Stockholm, Sweden. PATIENTS: 180 patients sick-listed for acute LBP. MAIN OUTCOME MEASURES: Direct, indirect and total costs for three treatment programmes. RESULTS: The direct costs for treatment were lowest in the GPP group -- 2744 Swedish crowns (SEK) per patient. More patients in the MTP and ITP underwent operations for disk hernia and radiological investigations than in the GPP. Indirect costs, defined as sick-leave for LBP represent about 90% of the total cost. CONCLUSIONS: With respect to total costs, the findings were similar between the three treatment programmes. The GPP had the lowest direct costs. It is not possible to conclude which treatment programme is to be recommended as a least cost alternative. The strong effect of indirect costs on the total cost stresses that further studies should focus on methods of shortening sick-leave. PMID- 10811046 TI - Prevalence of non-healed and healed chronic leg ulcers in an elderly rural population. AB - OBJECTIVE: To estimate the prevalence of non-healed and healed chronic leg ulcers in an elderly rural population. DESIGN: All persons 70 years or older were asked about present and previous leg ulcers. SETTING: A rural village in mid-west Sweden with 4000 inhabitants. PARTICIPANTS: 541 persons aged 70 years and above. OUTCOME MEASURES: The prevalence of healed or non-healed chronic leg ulcers in the population aged > or = 70. RESULTS: Healed or non-healed leg ulcers were reported by 53 persons (9.8%). Examination of these patients and also 100 individuals who had negated present or previous leg ulceration, giving the total prevalence of 12.6%. CONCLUSION: The result indicates a certain risk for overestimation of prevalence, as well as underestimation and decreased reliability when basing the figures only on telephone interviews or posted questionnaires. PMID- 10811047 TI - Poor regulation of diabetic patients: a common phenomenon? PMID- 10811048 TI - Implementing clinical guidelines in the treatment of hypertension. PMID- 10811049 TI - Hormonal fountains of youth. PMID- 10811050 TI - New insights into the the molecular mechanisms of alcoholic hepatitis: a potential role for NF-kappaB activation? PMID- 10811051 TI - Androgen deficiency in aging men: role of testosterone replacement therapy. PMID- 10811052 TI - Hypertension, hormones, and aging. AB - The most rapidly growing segment of the United States population is the geriatric group, especially those above 75 years of age. Hypertension, diabetes mellitus, and dyslipidemia increase with advancing years in Westernized, industrialized societies such as the United States. These disorders contribute significantly to strokes and myocardial infarctions and associated morbidity and mortality in our elderly population. The increase in these chronic disease processes with aging is related, in part, to increasing obesity, reductions in physical activity, and medications that predispose to these conditions (ie, nonsteroidal inflammatory agents and hypertension). Hypertension in the elderly is characterized by high peripheral vascular resistance/reduced cardiac output, impaired baroflex sensitivity, relatively greater systolic pressures, increased blood pressure variability, and a propensity to salt sensitivity. Type 2 diabetes in the elderly is related to alterations in body composition (ie, increased central adiposity and decreased lean body mass) and to reduced physical activity. There is an increasing body of evidence that aggressive treatment of hypertension and dyslipidemia in the elderly results in comparable, if not greater, reductions in cardiovascular morbidity and mortality in the elderly as in younger counterparts. PMID- 10811053 TI - Increased monocyte nuclear factor-kappaB activation and tumor necrosis factor production in alcoholic hepatitis. AB - Increased tumor necrosis factor-a activity has been reported in patients with alcoholic hepatitis and is implicated in its pathogenesis. The aim of this study was to investigate potential mechanisms of increased tumor necrosis factor-a activity in alcoholic hepatitis. Monocyte nuclear factor-kB activity was assessed by electrophoretic mobility shift assay, monocyte tumor necrosis factor-a mRNA was semi-quantitatively assessed by reverse transcriptase polymerase chain reaction, and tumor necrosis factor-a in monocyte culture supernatants was measured. There was significantly greater spontaneous nuclear factor-kB activity in the monocytes of 6 patients with alcoholic hepatitis as compared with that in the monocytes of control subjects. There was spontaneous tumor necrosis factor-a mRNA and tumor necrosis factor-a release from the monocytes of patients with alcoholic hepatitis but not from the monocytes of normal subjects. Endotoxin increased nuclear factor-kB activity and induced tumor necrosis factor-a mRNA and tumor necrosis factor-a release from normal subjects' monocytes. Endotoxin further increased nuclear factor-kB activity, tumor necrosis factor-a mRNA, and tumor necrosis factor-a release from the monocytes of patients with alcoholic hepatitis. Supershift assays indicate that the monocyte nuclear factor-kB activation involves the p50 and p65 subunits. Dysregulated tumor necrosis factor a metabolism in alcoholic hepatitis monocytes is associated with increased nuclear factor-kB activity and tumor necrosis factor-a mRNA expression. PMID- 10811054 TI - Blood glutathione and cysteine changes in cardiovascular disease. AB - The need to investigate aminothiols such as glutathione (GSH), cysteine (Cys), and homocysteine (Hcy) in blood is stimulated by the current interest in hyperhomocysteinemia as a risk factor for atherosclerosis. Our current goal was to determine whether various cardiovascular (CV) diseases altered levels of GSH and Cys in blood and the relationships between these two thiols. Blood samples from 96 patients with atherosclerosis and other CV diseases were analyzed and compared with those from 33 control subjects. In CV patients, GSH levels were normal, but free plasma Cys was significantly higher (P < .0001). In patients with atherosclerosis, bound plasma Cys was 21% higher than that in control subjects (P < .0001), and in patients with other CV diseases it was 14% higher (P = .023). Also, in patients with CV diseases, correlations of free GSH with free Cys (P < .007) and total GSH and Cys with age (P < .04) differed from that in control subjects. There were no differences related to functional disability or duration of disease. A key finding was that these abnormal levels of plasma Cys occurred in both atherosclerotic and non-atherosclerotic CV diseases. These results indicate that high levels of oxidized and bound Cys in CV patients create an oxidative environment that may increase susceptibility to vascular damage. PMID- 10811055 TI - Blood glutathione decreases in chronic diseases. AB - Previously a high blood glutathione level was correlated with long life span in the mouse and rat and in healthy elderly human beings. This raised the question of whether low glutathione levels occur in unhealthy subjects. To this end, 74 consecutive patients newly admitted to the hospital, with ages ranging from 21 to 89 years and diagnosed with chronic diseases, were studied along with 32 healthy control subjects. Blood samples were analyzed for reduced (GSH) and oxidized (GSSG) glutathione with a high-performance liquid chromatography-dual electrochemical method. The data were integrated with the clinical diagnoses and statistically analyzed. Marked total glutathione decreases from the control levels occurred in over 36% of the patients with chronic diseases including cancer and genitourinary, gastrointestinal, cardiovascular, and musculoskeletal diseases (P < .001). The deficit was due to low GSH concentrations and not to GSSG, which was the same as that in the control subjects. The conclusion is that a decrease in GSH is a risk factor for chronic diseases that may be used to monitor the severity and progress of the diseases. Future work is necessary to elucidate the mechanism of action. PMID- 10811056 TI - Platelet-derived growth factor receptor tyrosine kinase inhibitor AG1295 attenuates rat hepatic stellate cell growth. AB - Enhanced activity of receptor tyrosine kinases such as the platelet-derived growth factor-receptorbeta (PDGF-Rbeta) has been implicated as a contributing factor in the development of hepatic fibrosis. In this study we have used tyrosine kinase inhibitors of the tyrphostin class (AG1295) to specifically block autophosphorylation of PDGF-Rbeta and proliferation of rat hepatic stellate cells. We also examined the effect of AG1295 on the PDGF-BB-induced activation of the 44 kd and 42 kd mitogen-activated protein (MAP) kinase isoforms (p44mapk/p42mapk). Rat hepatic stellate cells were treated with AG1295 (10 micromol/L) for 24 hours and stimulated with PDGF-BB for 5 minutes. AG1295 specifically inhibited autophosphorylation of PDGF-Rbeta and caused a 20% decrease in PDGF-BB-stimulated bromodeoxyuridine incorporation by rat hepatic stellate cells. Treatment of rat hepatic stellate cells with AG1295 resulted in an inhibition of the PDGF-BB-induced activation of MAP kinase isoforms. Quantification of the immunoprecipitated tyrosine-phosphorylated phosphatidylinositol 3-kinase, phospholipase C-gamma, and p21ras guanosine triphosphatase-activating protein by Western blotting revealed that AG1295 treatment effectively inhibits tyrosine phosphorylation of these kinases in hepatic stellate cells. Our findings demonstrate that AG1295 is a selective inhibitor of the tyrosine phosphorylation of PDGF-Rbeta and its downstream signaling pathway, and this compound could offer a strategy for the treatment of fibrotic liver diseases. PMID- 10811057 TI - Serum ferritin iron in iron overload and liver damage: correlation to body iron stores and diagnostic relevance. AB - The iron content of serum ferritin has been determined in groups of patients with normal or increased iron stores by using a technique of ferritin immunoprecipitation followed by iron quantitation with atomic absorption spectroscopy. The results were correlated to individual liver iron concentrations, measured non-invasively by superconducting quantum interference device (SQUID) biomagnetometry. A close correlation between serum concentrations of ferritin protein and ferritin iron was found (r = 0.92) in all groups of patients. However, the correlation between ferritin iron concentration and individual liver iron concentration was poor in patients with hemochromatosis (r = 0.63) and patients with beta-thalassemia major (r = 0.57). The degree of ferritin iron saturation was about 5% in iron-loaded patients, which contrasts with results in two recent studies but confirms older observations. In patients with liver cell damage, the ferritin iron saturation in serum was significantly higher than that found in groups with iron overload disease, probably indicating the release of intracellular iron-rich ferritin into the blood. The monitoring of patients undergoing bone marrow transplantation indicated that the release of iron-rich and iron-poor ferritin occurred during phases of hepatocellular damage and inflammation, respectively. We find the benefits of serum ferritin iron measurement to be marginal in patients with iron overload disease. PMID- 10811058 TI - Decreased plasma concentrations of L-hydroxy-arginine as a marker of reduced NO formation in patients with combined cardiovascular risk factors. AB - Patients with metabolic syndrome represent a group with extensive cardiovascular risk factors for the development of atherosclerosis, which may be preceded by an impairment of endothelial function. Endothelial dysfunction is characterized by a reduced availability of bioactive nitric oxide, the principal mediator of endothelium-dependent vasodilation. In the present study we assessed NO synthesis in vivo by measuring the NO-related amino acids L-arginine and L-citrulline and in particular the stable intermediate compound N(omega)-hydroxy-L-arginine (L NHA) in patients with metabolic syndrome by using high-performance liquid chromatography (HPLC) analysis. As a prerequisite to our study, we measured the amino acid concentrations in 31 healthy volunteers to investigate gender and age differences. To prove whether blood drawn from peripheral veins reflects plasma concentrations of the whole vessel system, several blood samples from different regions were obtained from patients undergoing elective left and right heart catheterization. In the latter group, no significant differences were noted among the plasma concentrations between the different sample sites. In healthy volunteers, there were no significant differences in plasma concentrations of any one specific amino acid between males and females or age groups. The main finding of the study is that the intermediate product of NO synthesis, L-NHA, is significantly reduced in the plasma samples of patients with a metabolic syndrome as compared with samples from healthy control subjects. The plasma concentrations of the NO precursor L-arginine and the end product of NO synthesis, L-citrulline, were unchanged. In conclusion, our results suggest that plasma levels of L-NHA are independent of age and gender and are not different at various locations within the vascular system. In a group of patients at high risk for the development of atherosclerosis, we found reduced plasma concentrations of L-NHA, either caused by a decreased endothelial NO synthase activity or caused by an increased breakdown of L-NHA by pathways independent of NO synthase, resulting in a reduced availability of L-NHA for NO synthesis. PMID- 10811059 TI - Management of the retinal impact site after intraocular foreign body trauma. PMID- 10811060 TI - Systemic prednisolone prevents rebleeding in traumatic hyphema. PMID- 10811061 TI - Traumatic optic neuropathy. PMID- 10811062 TI - PRK for myopia and astigmatism. PMID- 10811063 TI - Glaucoma in patients with sleep apnea. PMID- 10811064 TI - National Survey of Local Anaesthesia for Ocular Surgery in the United Kingdom. PMID- 10811065 TI - Cat scratch disease: posterior segment manifestations. PMID- 10811066 TI - Simultaneous versus bilateral sequential LASIK. PMID- 10811067 TI - More on simultaneous versus sequential LASIK. PMID- 10811068 TI - Posterior segment intraocular foreign bodies: management in the vitrectomy era. PMID- 10811069 TI - Prognostic factors in ocular injuries caused by intraocular or retrobulbar foreign bodies. AB - OBJECTIVE: To evaluate prognostic factors associated with final visual outcome, development of posttraumatic infectious endophthalmitis, and occurrence of proliferative vitreoretinopathy in patients with penetrating ocular injuries caused by intraocular or retrobulbar foreign bodies (FBs). DESIGN: Clinic-based cross-sectional study. PARTICIPANTS: One hundred thirty patients presenting with penetrating ocular injuries caused by lacerations from FBs were operated on by one of two surgeons between 1989 and 1997. Follow-up time was an average of 20.84 +/- 20.76 months (median: 17.0 months). All FBs were located posterior to the lens. INTERVENTIONS: Pars plana vitrectomy; foreign body removal; additional surgical procedures according to the clinical situation. MAIN OUTCOME MEASURES: Postoperative visual acuity; posttraumatic infectious endophthalmitis; proliferative vitreoretinopathy. RESULTS: Occurrence of posttraumatic infectious endophthalmitis developing in seven patients (7/130 = 5.4%) was significantly (P = 0.026) associated with removal of the FB later than 24 hours after the accident and with the type of the FB (P < 0.01). Size (P = 0.37) of the FB, preoperative visual acuity (P = 0.62), presence of traumatic cataract (P = 0.75) or a retinal lesion by the FB (P = 0.16), age (P = 0.39), and gender (P = 0.46) did not show a statistically significant influence on the occurrence of endophthalmitis. Statistically significant risk factors for the development of proliferative vitreoretinopathy occurring in 27 patients (27 of 99 [27.6%] patients with a minimal follow-up of 3 months) were size of the FB (P < 0.001), preoperative visual acuity (P = 0.02), presence of a retinal lesion (P = 0.002), and traumatic cataract (P = 0.03). The time between FB removal and the accident was statistically marginally associated with the development of proliferative vitreoretinopathy (P = 0.07). Postoperative visual acuity depended significantly on size of the FB (P = 0.002), preoperative visual acuity (P < 0.001), presence of a retinal lesion (P = 0.049), and location of the retinal lesion (P < 0.001). Three eyes had to be enucleated because of endophthalmitis or phthisis bulbi. CONCLUSIONS: Prognosis in open-globe injuries with intraocular or retrobulbar foreign bodies depends on the size and type of the foreign body, presence and location of retinal lacerations, additional involvement of other intraocular structures, preoperative visual acuity, and timing of surgery. These factors may be important in preoperative counseling of the patient and for planning surgery. PMID- 10811070 TI - Internal orbital fractures in the pediatric age group: characterization and management. AB - OBJECTIVE: To evaluate the specific characteristics and management of internal orbital fractures in the pediatric population. DESIGN: Retrospective observational case series. PARTICIPANTS: Thirty-four pediatric patients between the ages of 1 and 18 years with internal orbital ("blowout") fractures. METHODS: Records of pediatric patients presenting with internal orbital fractures over a 5 year period were reviewed, including detailed preoperative and postoperative evaluations, surgical management, and medical management. MAIN OUTCOME MEASURES: Ocular motility restriction, enophthalmos, nausea and vomiting, and postoperative complications. RESULTS: Floor fractures were by far the most common fracture type (71%). Eleven of 34 patients required surgical intervention for ocular motility restriction. Eight were trapdoor-type fractures with soft-tissue incarceration; five had nausea and vomiting. Early surgical intervention (<2 weeks) resulted in a more complete return of ocular motility compared with the late intervention group. CONCLUSIONS: Trapdoor-type fractures, usually involving the orbital floor, are common in the pediatric age group. These fractures may be small with minimal soft-tissue incarceration, making the findings on computed tomography scans quite subtle at times. Marked motility restriction and nausea/vomiting should alert the physician to the possibility of a trapdoor-type fracture and the need for prompt surgical intervention. PMID- 10811071 TI - Ocular injuries sustained by survivors of the Oklahoma City bombing. AB - OBJECTIVE: The purpose of this study is to provide a review of the ocular injuries sustained by survivors of the April 19, 1995, bombing of the Alfred P. Murrah Federal Building in Oklahoma City. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: The authors retrospectively evaluated data collected on all surviving persons receiving ocular injuries during the bombing and on all at-risk occupants of the federal building and four adjacent buildings. METHODS: Injury data from survivors were collected from multiple sources to include hospital medical records, a physician survey, emergency medical services run reports, written survivor accounts, building occupant survey, telephone interviews, and mail surveys. MAIN OUTCOME MEASURES: The types of ocular injuries, the associated systemic injuries, and the location of the injured at the time of the blast were evaluated. RESULTS: Fifty-five (8%) of the 684 injured bombing survivors sustained an ocular injury. Persons injured in the Murrah building were more than three times more likely to sustain an ocular injury than other injured persons. Seventy-one percent of ocular injuries occurred within 300 feet of the point of detonation. The most common serious ocular injuries included lid/brow lacerations (20 patients, 23 eyes), open globe injuries (12 eyes), orbital fractures (6 eyes), and retinal detachment (5 eyes). A retained intraocular foreign body accounted for only two of the injuries (4%). Glass accounted for nearly two thirds of the ocular injuries. CONCLUSIONS: Blasts involving explosions inflict severe ocular injury, mostly as a result of secondary blast effects from glass, debris, etc. Eye injuries in bombings can probably be prevented by increasing the distance from and orientation away from windows (i.e., by facing desks away from windows). Use of such products as laminated glass, toughened window glazing, and Mylar curtains may reduce glass projectiles in the blast vicinity. PMID- 10811072 TI - Eye injuries in a terrorist bombing: Dhahran, Saudi Arabia, June 25, 1996. AB - OBJECTIVE: We report the experience of our institution in the evaluation and care of multiple simultaneous ocular trauma patients after a terrorist bomb attack on a United States military base in Saudi Arabia. DESIGN: Retrospective, noncomparative small case series. PARTICIPANTS: Three patients who received severe ocular injuries after a terrorist bombing. INTERVENTION: All patients underwent surgical repair of the injuries that were inflicted as a result of the terrorist bombing. MAIN OUTCOME MEASURES: Baseline ocular characteristics, intraoperative findings, surgical procedures, and final (3 years after injury) anatomic and visual outcomes were noted. RESULTS: Glass fragments caused by the blast were the mechanism of all the ocular injuries in these patients. All patients had primary repair of the injuries done in Saudi Arabia and were sent to our institution for tertiary care. Three of the four eyes injured had stable or improved visual acuity and one eye was enucleated. Two patients had no serious injury other than the globe trauma. One patient had extensive eyelid trauma and required serial procedures to allow fitting of a prosthesis. CONCLUSIONS: Blast injury patients are at risk for open globe injury as a result of glass fragments. The types of injury that can occur from terrorist blasts can be extensive and involve all the tissues of the eye, the ocular adnexa, and the orbit. PMID- 10811073 TI - Scleral rupture during retinal detachment surgery: risk factors, management options, and outcomes. AB - OBJECTIVE: To evaluate the potential risk factors, management strategies, and outcomes of scleral rupture during retinal detachment (RD) surgery. DESIGN: Case control study. PARTICIPANTS AND CONTROLS: Fourteen consecutive patients with scleral rupture during RD surgery (cases) and 65 consecutive patients who underwent RD surgery without scleral rupture (controls). INTERVENTION: Demographic and clinical data were abstracted from patients' medical records. OUTCOME MEASURES: Visual acuity and retinal attachment status at the last examination. RESULTS: Significant risk factors for scleral rupture during RD surgery were reoperation after failed RD surgery (71 % vs. 32%), and pre-existing scleral pathologic condition (29% vs. none). The site of rupture was in the bed of a previously placed scleral buckle in all patients with a previous buckling surgery. Repair of the rupture included scleral sutures in eight (57%), scleral patch graft in four (29%), and placement of a scleral buckle over the site in two (14%) eyes. Eleven (79%) underwent vitrectomy with retinal tamponade by gas (n = 4) or silicone oil (n = 7). Complications observed postoperatively included vitreoretinal incarceration (n = 3), vitreous hemorrhage (n = 2), suprachoroidal hemorrhage (n = 2) and subretinal hemorrhage (n = 3). In the 14 eyes with scleral rupture, the final visual acuity was > or =20/40 in 1 (7%), 20/50 to 20/200 in 5 (36%), and <20/200 in 8 (57%). Ten (71 %) had proliferative vitreoretinopathy develop. The retina was attached in 7 (50%), 6 (43%) had localized peripheral detachment, and 1 had a total retinal detachment. The vision improved in 4 (29%), was unchanged in 5 (36%), and was worse than before surgery in 5 (36%). In the 65 controls, the visual acuity at the time of the last examination was > or =20/40 in 26 (40%), 20/50 to 20/200 in 21 (32%), and <20/200 in 18 (28%). Sixty three (97%) patients had complete retinal reattachment, 1 (2%) had a localized peripheral RD, and 1 (2%) had an RD involving the posterior pole. After surgery, the vision improved in 45 (69%), was unchanged in 15 (23%), and was worse in 5 (8%) of the control eyes. The visual and anatomic outcomes of the eyes with scleral rupture were significantly worse than in the control group (P = 0.002 and P < 0.001, respectively). CONCLUSIONS: Risk factors associated with intraoperative scleral rupture include reoperation for failed RD surgery and pre existing scleral pathology. Although this complication may be compatible with a good visual outcome in some patients, a high incidence of persistent or recurrent RD with proliferative vitreoretinopathy worsens the visual outcome for most patients with this complication. PMID- 10811074 TI - Prevalence, correlates, and natural history of epiretinal membranes surrounding idiopathic macular holes. Virectomy for Macular Hole Study Group. AB - OBJECTIVE: To report on the prevalence, correlates, and natural history of epiretinal membranes (ERM) in eyes with stage II or III/IV macular holes. DESIGN: A subgroup analysis arising from a multicentered, controlled, randomized clinical trial. SETTING: Community and university-based ophthalmology clinics. PATIENTS: Two hundred twenty four eyes with stage II or III/IV macular holes. INTERVENTIONS: No intervention for 100 eyes randomly assigned to observation. OUTCOME MEASURES: Grade of ERM determined by stereoscopic examination of fundus photographs at baseline and at 3, 6, 12, and 24 months of follow-up. RESULTS: The prevalence of ERM was 65% (145 of 224 eyes), was greater in pseudophakic than in phakic eyes (80% vs. 63%, P = 0.10), and increased with increasing severity of the hole (P < 0.0001). Stage III/IV eyes with ERM had a significantly larger hole size than did eyes without ERM (P < 0.01); however, no association between presence of ERM and visual acuity was found (P > 0.5). In the 100 phakic eyes that were randomly assigned to observation, there was a significant increase in the severity of ERM over follow-up (P < 0.0001). CONCLUSIONS: ERM are common in eyes with full-thickness idiopathic macular holes. Although ERM prevalence increases with severity and size of the macular hole, the presence of ERM are not closely correlated with visual acuity. These factors may be important in considering the removal of ERM during vitrectomy for macular hole. PMID- 10811075 TI - Outcomes and complications associated with perfluoro-n-octane and perfluoroperhydrophenanthrene in complex retinal detachment repair. AB - OBJECTIVE: To compare rates of perfluorocarbon liquid (PFCL) intraocular retention, anatomic and visual acuity outcomes, and complications associated with intraoperative perfluoro-n-octane (Perfluoron) versus perfluoroperhydrophenanthrene (Vitreon) in retinal detachment repair. DESIGN: Retrospective noncomparative consecutive case series. PARTICIPANTS AND METHODS: Records of consecutive patients who underwent retinal detachment repair with intraoperative Perfluoron (n = 78) or Vitreon (n = 84) at the Bascom Palmer Eye Institute between November 1, 1991, and October 31, 1994, were retrospectively reviewed. MAIN OUTCOME MEASURES: Comparison of PFCL intraocular retention rates, retinal reattachment rates, visual acuity outcomes, and postoperative complication rates between the Perfluoron and Vitreon groups at postoperative day 1, week 1, month 1, month 3, and month 6. To assess the study's generalizability, the Perfluoron data obtained in the current study were compared with results of the Perfluoron Multicenter Clinical Study. RESULTS: Retained Perfluoron was noted less frequently (P < 0.03) than retained Vitreon at each postoperative visit assessed. At 6 months postoperatively, the cumulative rate of retained PFCL (noted at any of the study postoperative visits) was 7.8% of patients in the Perfluoron group and 38.3% in the Vitreon group (P < 0.001). No significant difference was found between the groups in retinal reattachment rates; at 6 months postoperatively, the retina was attached in 45 of 61 (74%) patients in the Perfluoron group and 36 of 57 (63%) patients in the Vitreon group (P = 0.2). There was a trend (P = 0.055) toward better 6-month visual acuity in the Perfluoron group compared with the Vitreon group. The incidence of corneal abnormality at any study visit was 46.7% in the Perfluoron group and 77.4% in the Vitreon group (P < 0.001). At 6 months postoperatively, the cumulative rate of elevated intraocular pressure (>25 mmHg) was 13 +/- 4% and 37 +/- 5%, respectively (P = 0.004). No significant difference was found between the groups in rates of postoperative hypotony. Results of this study are comparable with those observed in the Perfluoron Multicenter Clinical Study. CONCLUSIONS: In this study, Perfluoron was retained intraocularly less frequently than Vitreon. Although no significant difference was found between groups in retinal reattachment rates, Perfluoron is associated with slightly better 6-month visual acuity and lower rates of corneal abnormality and elevated intraocular pressure compared with Vitreon. PMID- 10811076 TI - A comparison of digital nonmydriatic fundus imaging with standard 35-millimeter slides for diabetic retinopathy. AB - PURPOSE: The purpose of this study was to compare nonmydriatic digital images with 35-mm slide images for the detection of diabetic retinal findings. STUDY DESIGN: Comparative, observational case series. PARTICIPANTS: Twenty-two patients with diabetes. METHODS: Diabetic patients underwent digital (nonmydriatic fundus camera attached to a digital back) and standard dilated 35-mm retinal photography of three areas: posterior pole, nasal retina, and temporal retina. The images were reviewed by a single masked grader for the presence or absence of specific retinal findings. PRIMARY OUTCOME MEASURES: Presence or absence of neovascularization of the disc (NVD), neovascularization elsewhere (NVE), venous beading (VB), nerve fiber layer (NFL) hemorrhage, dot-blot hemorrhage, microaneurysm (MA), clinically significant macular edema, cotton wool spot, intraretinal microvascular anomaly (IRMA), hard exudate (HE), and retinal pigment epithelial (RPE) pigmentary changes. RESULTS: Forty eyes of 22 patients underwent both imaging procedures. The agreement between image type was highest for IRMA (97.5%) and VB (95%) and lowest for RPE pigmentary changes (65%) and MA (62.5%). Sensitivity ranged from 25% (NVD) to 100% (VB). Specificity ranged from 90% (RPE pigmentary changes) to 100% (NVD, NVE, VB, NFL hemorrhage, HE). Positive predictive value ranged from 50% (IRMA) to 100% (NVD, NVE, HE, NFL, VB). Negative predictive value ranged from 48% (MA) to 100% (IRMA). CONCLUSIONS: Nonmydriatic digital fundus imaging for detection of diabetic retionopathy has a low sensitivity rate and a high specificity rate and is less clinically useful than standard dilated 35-mm fundus slide images. PMID- 10811077 TI - Prevalence of serologic evidence of cat scratch disease in patients with neuroretinitis. AB - OBJECTIVE: To determine the prevalence of Bartonella henselae seropositivity in patients with a clinical diagnosis of neuroretinitis. DESIGN: Retrospective, clinic-based, cross-sectional study. PARTICIPANTS: Eighteen consecutive patients seeking treatment at the Casey Eye Institute from November 1993 through November 1998 who had neuroretinitis. METHODS: The billing and photographic records of the Casey Eye Institute were searched for patients with a primary or secondary diagnosis of neuroretinitis or Leber's idiopathic stellate neuroretinitis. Charts were then reviewed to determine the results of B. henselae antibody titers and other pertinent clinical information. MAIN OUTCOME MEASURES: Results of B. henselae serologic testing. RESULTS: Fourteen of 18 patients with neuroretinitis had serologic studies. Nine of the 14 tested patients (64.3%) were found to have elevated IgM or IgG for B. henselae, suggesting current or past infection. Patients with positive serologic analysis results tended to have worse vision at presentation. There were no other obvious differences between seropositive and seronegative groups in this study, including duration or quality of recovery. CONCLUSIONS: At our tertiary care ophthalmology institution, most tested patients with neuroretinitis had evidence of past or present cat-scratch disease based on positive serologic analysis for B. henselae, a much greater prevalence than is expected to be found in the general population or in patients with idiopathic uveitis. Further study is indicated to clarify the prevalence of cat-scratch disease in neuroretinitis and the role and efficacy of antibiotics in treatment. PMID- 10811078 TI - Highly active antiretroviral therapy-related immune recovery in AIDS patients with cytomegalovirus retinitis. AB - OBJECTIVE: To characterize cytomegalovirus (CMV) retinitis in human immunodeficiency virus (HIV)-infected patients who demonstrate immune recovery while receiving highly active antiretroviral therapy (HAART). DESIGN: Consecutive, noncomparative case series. PARTICIPANTS: Twenty-two HIV-positive patients, from two institutions, with a history of CMV retinitis, and with elevated CD4 cell counts after HAART. MAIN OUTCOME MEASURES: Duration of healed CMV retinitis without anti-CMV therapy, CD4 cell count, and HIV viral load. INTERVENTION: Discontinuation of anti-CMV therapy after persistent elevation of CD4 cell count over 50 cell/mm3 (median, 161/mm3; range, 85-408/mm3). RESULTS: The median period of healed CMV retinitis without anti-CMV therapy was 72 weeks (range, 33-116 weeks). Nineteen of 22 patients were still healed without anti-CMV therapy at study end. The three patients with CMV retinitis progression simultaneously had HAART, fail with CD4 cell counts of 37, 35, and 47/mm3. CONCLUSIONS: HIV-positive patients with CMV retinitis, who demonstrate a sustained HAART-induced elevation of CD4 cell count on two consecutive counts 3 months apart and whose retinitis remains healed on anti-CMV therapy for greater than 4 months, are likely to remain healed if the anti-CMV therapy is withdrawn. It is important to monitor these patients with indirect ophthalmoscopy because HAART failure may occur and allow CMV retinitis reactivation. PMID- 10811079 TI - Color vision in epilepsy patients treated with vigabatrin or carbamazepine monotherapy. AB - PURPOSE: To investigate color vision in epilepsy patients treated with vigabatrin or carbamazepine monotherapy and to evaluate the association between vigabatrin induced visual field defects and dyschromatopsia. DESIGN: Nonrandomized comparative trial. PARTICIPANTS: Thirty-two epilepsy patients treated with vigabatrin monotherapy, 18 patients treated with carbamazepine monotherapy, and 47 age-matched healthy controls were examined. MAIN OUTCOME MEASURES: Color vision was examined with Standard Pseudoisochromatic Plates 2 (SPP2) screening test, Farnsworth-Munsell 100 (FM 100) hue test, and Color Vision Meter 712 anomaloscope. RESULTS: Abnormal color perception was found in 32% of the epilepsy patients treated with vigabatrin monotherapy and 28% of the epilepsy patients treated with carbamazepine monotherapy. The total error score in the Farnsworth Munsell 100 hue test was abnormally high in the vigabatrin monotherapy patients who had concentrically constricted visual fields and a statistically significant correlation was found between the temporal visual field extents and the age adjusted Farnsworth-Munsell 100 total error score in vigabatrin monotherapy patients (R = .533, P = 0.003 in the right eye, R = .563, P = 0.001 in the left eye). Four of 31 (12%) vigabatrin monotherapy patients, and 1 of 18 (6%) carbamazepine monotherapy patients had a blue axis in Farnsworth-Munsell 100 hue test. In the anomaloscope, there were a few pathologic findings in both groups. In the SPP2 screening test, a few plates were not seen in both groups. CONCLUSIONS: Both examined antiepileptic drugs, vigabatrin and carbamazepine, cause acquired color vision defects. The abnormal color perception seems to be associated with constricted visual fields in the vigabatrin monotherapy patients. The duration of carbamazepine therapy correlates with high FM100 total error score. The best method for detecting dyschromatopsia in patients treated with vigabatrin or carbamazepine was the Farnsworth-Munsell 100 hue test. The SPP2 screening test does not seem to be useful in clinical practice. PMID- 10811080 TI - Rand-Stein analgesia protocol for cataract surgery. AB - OBJECTIVE: To describe the safety and efficacy of an analgesia protocol that enables the surgeon to maintain control over an alert patient experiencing seemingly painless ambulatory cataract surgery, while eliminating the risks and side effects associated with general, local, topical, and intracameral anesthesia. DESIGN: Noncomparative, interventional case series. PARTICIPANTS: Five thousand one hundred sixty-eight consecutive cataract surgery cases operated on by the same surgeon from April 1, 1993 through June 1, 1998. METHODS: This technique produces profound ocular analgesia, avoiding any undesired sedative effects, using very low-dose, titrated, intravenous alfentanil. Complete control of the uncooperative patient, including lid squeezing and ocular and general body movements, is obtainable whenever necessary using very low-dose, titrated, intravenous methohexital. MAIN OUTCOME MEASURES: Success was defined as surgery completed in a controlled manner without the need to convert to general, local, topical, or intracameral anesthesia and the patient's experience being perceived as pain free. RESULTS: One hundred percent of the cases were successful without ever deviating from the protocol. CONCLUSIONS: This analgesia protocol offers advantages for cataract surgery. It virtually eliminates the morbidity of cataract surgery associated with other anesthesia techniques while providing excellent and reliable control. It allows for an immediate postoperative recovery with instantaneous vision restoration. These patients are generally awake, alert, and retain their protective reflexes. PMID- 10811081 TI - Intraocular lidocaine in phacoemulsification: an endothelium and blood-aqueous barrier permeability study. AB - OBJECTIVE: To evaluate the effects of intraocular lidocaine hydrochloride (HCI) on the corneal endothelium and of blood-aqueous barrier (BAB) permeability in ultrasound phacoemulsification. DESIGN: Nonrandomized, comparative trial. PARTICIPANTS: Sixty patients who underwent uneventful, single-surgeon, clear cornea phacoemulsification were studied prospectively. METHODS: Sub-Tenon's anesthesia was administered to 30 patients (group 1), and intraocular lidocaine hydrochloride was administered to an additional 30 patients (group 2). An endothelial study of at least 120 cells per patient, using a noncontact specular microscope and a digital image analysis system, was performed before surgery and 1 month after surgery. Blood-aqueous barrier permeability was evaluated preoperatively and postoperatively (48 hours, 1 week and 1 month) using a laser flare meter. MAIN OUTCOME MEASURES: Endothelial parameters (cell density, hexagonality, and coefficient of variation in cell area) and BAB permeability. RESULTS: No significant differences were found between groups when comparing the postoperative changes produced in endothelial cell density (95% confidence interval [CI], 0.46, 4.7; P = 0.10), hexagonality (95% CI, -2.5, 3.5; P = 0.72), and coefficient of variation in cell area (95% CI, -8.3, 4.6; P = 0.57). A multivariate study detected no significant differences in mean flare values between groups during follow-up (P = 0.40). No clinically significant differences were found between mean preoperative and 1 month postoperative flare values in either group. CONCLUSIONS: The recovery of BAB permeability 1 month after surgery seems to suggest that no additional inflammation was induced by intraocular lidocaine HCI. The fact that no differences in postoperative changes in endothelial parameters were found between groups indicates that no further corneal endothelial damage (other than that caused by surgical manipulation) was incurred. According to our results, intracameral lidocaine HCI appears to be safe for the average phacoemulsification patient in the absence of ocular pathologic conditions. PMID- 10811082 TI - Ultrasonographic measurement of induced myopia associated with capsular bag distention syndrome. AB - OBJECTIVE: To determine the causes of postoperative myopia associated with capsular bag distention syndrome (CBDS) and characterize the associated findings. DESIGN: Prospective, multicenter, nonrandomized, comparative (self-controlled) trial. PARTICIPANTS: Six eyes from six patients had CBDS develop after phacoemulsification and posterior chamber intraocular lens implantation. METHODS: Keratometric values, axial length, visual acuity, and manifest refraction were assessed in each eye. Using 20-MHz (I3SYSTEM-ABD, Innovative Imaging Inc, Sacramento, CA) and 50-MHz (Ultrasound Biomicroscope, Zeiss Humphrey Systems, Dublin, CA) ultrasonographic probes, images and measurements of the anterior segment were obtained. After neodymium:yttrium-aluminum-garnet (Nd:YAG) capsulotomy, these measures were repeated and correlated with predicted and actual refractive changes. MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), manifest refraction, and anterior chamber depth before and after treatment of CBDS. RESULTS: Although intended postoperative refraction averaged -0.58 diopters (D) (range, -0.12 to -1.63 D), eyes with CBDS had an average spherical equivalent refraction of -2.35 D (range, +0.13 to -4.50 D), P < 0.05 (one-tailed, paired t test). BCVA averaged 20/24 (range, 20/15-20/40-1), but UCVA averaged 20/133 (range, 20/60 to 20/400). Average distance from the corneal surface to the anterior intraocular lens (IOL) optic surface was 3.55 mm in eyes with CBDS, and 4.30 mm after Nd:YAG capsulotomy. Posterior movement of the IOL optic after capsulotomy accounted for 1.23 D of hyperopic shift or 82% of the CBDS-induced myopia. Treatment of CBDS resulted in both improved UCVA and BCVA. Nd:YAG capsulotomy also released the colloidal suspension within the capsular bag posterior to the IOL optic. The incidence of CBDS was 0.3% in one of the practices reviewed. CONCLUSIONS: Capsular bag distention syndrome includes unexpected myopia and poor UCVA after cataract surgery with lens implantation in cases involving a continuous capsulorhexis. A dramatic posterior distention of the posterior capsule is observed, as well as anterior chamber shallowing, tight apposition of the iris to the IOL, and anterior bowing of the iris. A slightly turbid colloidal suspension behind the IOL implant and late posterior capsular fibrosis are also observed. Timely treatment of CBDS can correct unwanted myopia, improve UCVA and BCVA, and restore normal anatomic relationships in the eye. PMID- 10811083 TI - Evaluation of foldable intraocular lenses in patients with uveitis. AB - OBJECTIVE: To evaluate various foldable posterior chamber intraocular lenses (IOLs) after phacoemulsification in patients with uveitis. DESIGN: A prospective, noncomparative, interventional case series. PARTICIPANTS: Forty-nine consecutive patients (60 eyes) with various types of uveitis (anterior, n = 20; posterior, n = 1; panuveitis, n = 37, intermediate, n = 2). INTERVENTION: All patients underwent phacoemulsification with foldable posterior chamber IOL implantation. All eyes were free of active inflammation at the time of surgery. A variety of IOL biomaterials were implanted: acrylic (n = 30), silicone (n = 17), and hydrogel (n = 13). MAIN OUTCOME MEASURES: Detailed examination was performed by one masked observer. Several parameters were compared for each implant biomaterial, including level of best corrected Snellen visual acuity at final follow-up, presence of posterior synechiae, anterior capsular phimosis, posterior capsule opacification, and the degree of cellular deposits on the IOL optic. RESULTS: There were 26 males and 23 females, aged 9 to 83 years (mean, 48 years). Follow-up ranged from 1 to 33 months (mean, 17.03 months). At final follow-up, 56 eyes (93.3%) had an improvement in visual acuity compared with preoperative levels as follows: 34 eyes (56.6%) achieved an improvement of four or more Snellen lines, and 44 eyes (73.3%) achieved 20/30 or better. Giant cells, observed on the IOL optic in 19 eyes (31.7%), were most often seen on the acrylic biomaterial at the 1-month follow-up, although this was not found to be statistically significant. Scratch marks produced by the lens-introducing forceps were seen in 24 eyes (40.0%), mainly on the acrylic and hydrogel optics. Posterior capsule opacification (PCO) occurred in 49 eyes (81.7%), with only 5 eyes requiring laser capsulotomy. There was no association between PCO and the various lens biomaterials. Other causes for reduced visual acuity included glaucomatous optic neuropathy (n = 5) and cystoid macular edema (n = 8). CONCLUSIONS: The use of foldable IOLs in eyes with uveitis is safe, but the optimal biomaterial has yet to be found. PMID- 10811084 TI - Microkeratome complications of laser in situ keratomileusis. AB - PURPOSE: To evaluate the incidence, types, and outcome of microkeratome complications that occur during laser in situ keratomileusis (LASIK). DESIGN: Retrospective, noncomparative, case series. PARTICIPANTS: Three thousand nine hundred ninety-eight eyes that underwent primary LASIK by four surgeons between November 1996 and August 1998 at a university-based refractive center. METHODS: All cases with significant microkeratome complications leading to abandonment of the LASIK procedure were identified and reviewed. MAIN OUTCOME MEASURES: Incidence of complications, change in best corrected visual acuity (BCVA), change in refractive error, and types of complication. RESULTS: There were 27 complications leading to abandonment of the LASIK procedure of 3998 eyes. The overall rate of microkeratome complication was 1 in 150 (0.68%), but it was 1 in 77 (1.3%) in the surgeons' first 1000 eyes, decreasing to 1 in 250 (0.4%) in the last 1000 eyes. Of the 24 planned bilateral cases, 15 complications (63%) happened on the first operated eye. Twenty-six of 27 eyes (96%) recovered to within one line of preoperative BCVA, and one eye lost two lines. At last examination before any repeat refractive procedures, spherical equivalent manifest refraction returned to within 1 diopter (D) of its preoperative value in 18 of 19 eyes (95%), and astigmatism in 16 of 19 eyes (84%) returned to within 1 D of its preoperative value. Sixteen of 27 eyes (59%) had repeat LASIK. Two eyes had complications at repeat LASIK, one of which led to abandonment of the LASIK procedure for a second time. CONCLUSIONS: There is a significant learning curve in the use of the microkeratome. If ablation is not performed, flap complications rarely lead to significant visual loss and generally do not result in a change in refractive error. PMID- 10811085 TI - Photorefractive keratectomy versus laser in situ keratomileusis: comparison of optical side effects. Summit PRK-LASIK Study Group. AB - OBJECTIVE: This report presents patient-reported optical symptoms after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). DESIGN: Preoperative and postoperative patient surveys in a prospective, multicenter, randomized clinical trial. PARTICIPANTS: Two hundred twenty eyes of 220 patients entered the study; 105 were randomized to PRK and 115 were randomized to LASIK. INTERVENTION: All patients received a one-pass, multizone excimer laser ablation as part of either a PRK or LASIK procedure. Attempted corrections ranged from 6.00 to 15.00 diopters (D). MAIN OUTCOME MEASURES: Glare, halo, and monocular diplopia symptoms as reported by patients on questionnaires before surgery and at the 6-month follow-up. Comparison was made between symptoms when using optical correction before surgery and symptoms without correction after surgery. RESULTS: For both the PRK and LASIK groups analyzed individually, the difference in average glare index before surgery and after surgery was not statistically significant (P = 0.54 for PRK; P = 0.15 for LASIK; t test). Twenty four PRK patients (41.4%) reported worsening of glare symptoms from baseline compared with 11 LASIK patients (21.6%); however, the difference between the two groups was not statistically significant (P = 0.086, chi-square test). Within the PRK group, the difference in average halo index before and after surgery was statistically significant (P = 0.0003, t test); in the LASIK group, it was not statistically significant (P = 0.1 1, t test). Thirty-four PRK patients (58.6%) reported worsening of halo symptoms from baseline compared with 26 LASIK patients (50.0%); this difference was not statistically significant (P = 0.086, chi-square test). For both the PRK and LASIK groups, the difference in average diplopia index before and after surgery was statistically significant (P < 0.0001 for PRK; 0.047 for LASIK; t test). Twenty-six PRK patients (44.8%) reported a worsening of monocular diplopia symptoms from baseline compared with 19 LASIK patients (35.8%); this difference was not statistically significant (P = 0.39, chi-square test). When changes in glare and halo from before surgery to after surgery were pooled as a glare-halo index, however, the PRK group did show a significantly greater likelihood of demonstrating an increase in symptoms compared with the LASIK group (P = 0.048, chi-square test). CONCLUSIONS: Optical sequelae of glare, halo, and monocular diplopia may occur in some patients after either both PRK or LASIK for moderate to high myopia; in contradistinction, many other patients' preoperative symptoms improve after surgery. On average, PRK patients show an increase in halo and diplopia symptoms, but not glare, after surgery, and LASIK patients show an increase in diplopia, but not glare and halo symptoms. There is a suggestion of a somewhat lesser tendency toward postoperative optical symptoms in LASIK compared with PRK treated eyes. PMID- 10811086 TI - Scanning electron microscopic characteristics of phakic intraocular lenses. AB - OBJECTIVE: To analyze the surface quality of new generation phakic intraocular lenses (IOLs). DESIGN: Experimental materials study. MATERIALS: Three different new generation phakic IOLs: angle-fixated anterior chamber lens Chiron Vision NuVita MA20 (polymethylmethacrylate [PMMAD, iris-fixated anterior chamber lens Ophtec Artisan Iris-Claw (PMMA), posterior chamber lens Staar ICM (polymer from porcine collagen and 2-hydroxyethyl methacrylate [HEMA]). METHODS: Representative samples of three different phakic IOLs underwent surface and edge-finish examination with light microscopy (LM). The phakic IOLs were then examined by use of scanning electron microscopy (SEM), and particular attention was given to optic surface quality, edge finish, haptic, and optic/haptic junction. RESULTS: In all IOLs the LM examination showed a smooth and homogeneous surface. No irregularities, particularly at the optic front and back surface, optic edge, haptic, and the optic/haptic junctions, were detected by SEM. One exception was a minor surface roughness at the claws of an Artisan iris-fixated anterior chamber IOL. CONCLUSIONS: Phakic IOLs are implanted either in the anterior or posterior chamber of healthy eyes, and high standards for their surface quality are required. The evaluation of surface properties with LM and SEM did not reveal any defects that contraindicate the implantation of phakic IOLs. PMID- 10811087 TI - Exposure of primary orbital implants in postenucleation retinoblastoma patients. AB - PURPOSE: To determine significant factors influencing the exposure of primary orbital implants in patients with retinoblastoma. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: One hundred nine consecutive patients (110 sockets) who had undergone enucleation for retinoblastoma from January 1993 to December 1997. METHODS: Two patients with recurrence of orbital retinoblastoma were excluded from further analysis, leaving 107 patients (108 sockets). The parameters analyzed included the patient's age; gender; ocular diagnosis; surgeon; type, covering, and size of the implant; the use of chemotherapy or radiotherapy; and the timing of these treatments in relation to enucleation. Study patients were divided into two main groups: the "treated group"-patients who had undergone adjuvant external beam radiotherapy or chemotherapy, and the "untreated group"-patients had undergone enucleation with or without cryotherapy, laser thermotherapy, or brachytherapy to the index or fellow eye. The following additional parameters were noted in the patients with exposed implants: time to exposure from date of enucleation and treatment of exposure. MAIN OUTCOME MEASURE: Exposure of orbital implants. RESULTS: There were two exposures caused by orbital recurrence of retinoblastoma. The rate of nontumor recurrence exposure was 28% (30 of 108). The median time to exposure was 136 days (range, 1-630 days). There were 18 exposures (35%,18 of 51) in the treated group, with a 34% exposure rate (13 of 38) in the chemotherapy group. The exposure rate was 21% (12 of 57) in the untreated group. The rates of exposure according to implant were: Vicryl mesh-wrapped hydroxyapatite (2 of 18, 11%), Medpor (8 of 13, 53%), plain polymethylmethacrylate (PMMA) (4 of 50, 8%), Mersilene-wrapped PMMA (9 of 17, 53%) and Castroviejo (7 of 10, 70%). Eight of the exposures (27%) were managed conservatively; the remainder required surgical repair. CONCLUSIONS: Results suggested that implant type and covering (P = 0.000) had a highly significant effect on the rate of exposure in postenucleation retinoblastoma patients. There was no statistical evidence that age, gender, ocular diagnosis, surgeon, size of the implant, or radiotherapy had an effect on implant exposure. There was an increased rate of exposure in the chemotherapy group, although this did not achieve statistical significance (P = 0.058), but a detrimental effect could not be excluded. PMID- 10811088 TI - Delayed orbital infection after endoscopic orbital decompression for dysthyroid orbitopathy. AB - OBJECTIVE: To present a delayed complication of endoscopic orbital decompression that has not been reported previously in the literature. DESIGN: Retrospective non-comparative small case series. PARTICIPANTS: Three patients with dysthyroid orbitopathy. INTERVENTION: The medical records of patients with dysthyroid orbitopathy who underwent endoscopic orbital decompression and subsequently developed orbital infection were reviewed. RESULTS: Three patients with dysthyroid orbitopathy developed orbital infection (cellulitis or abscess) originating from the frontal sinus more than 2 years after their endoscopic orbital decompression surgery. Management required drainage of the abscess, administration of antibiotics, and creation of adequate frontal sinus drainage. CONCLUSIONS: Delayed orbital infection can occur after endoscopic orbital decompression for dysthyroid orbitopathy when the frontal sinus ostium is obstructed by orbital fat or scar tissue. Infection within the frontal sinus can cause secondary orbital cellulitis or abscess. Early signs and symptoms of a frontal sinus infection can be easily misdiagnosed as progression of the patient's thyroid eye disease. Awareness of this possible complication followed by appropriate early intervention will prevent a potentially blinding condition. Furthermore, ever since this complication was observed, the authors' surgical technique of endoscopic decompression has been modified to leave the most anterosuperior portion of the lamina papyracea to prevent fat prolapse and scar formation into the region of the frontal recess. PMID- 10811089 TI - Malignant melanoma of the eyelid and palpebral conjunctiva treated with iodine 125 brachytherapy. AB - OBJECTIVE: To evaluate the results on malignant melanomas of the eyelid and palpebral conjunctiva treated with iodine-125 (125I) brachytherapy with shielding of the eye. DESIGN: Noncomparative case series. PARTICIPANTS: Fourteen patients treated with 125I brachytherapy at an ocular oncology referral center from 1974 through 1996, all of whom had had previous debulking, incomplete resection, or recurrence after surgery. INTERVENTION: A stainless steel pericorneal ring eyeshield was attached to the extraocular muscles, over which a lid was fitted to protect the cornea without touching it. Iodine-125 seeds in polythene tubes were inserted into the eyelid and attached to the lid margin for a single plane implant. In five cases additional seeds were glued on to the shield as well for a volume implant. A median dose of 37 Gy (range, 17.3-67.6 Gy) was given over the course of 113 hours (range, 47-190 hours) to the outer surface of target volume. MAIN OUTCOME MEASURES: Local control is regarded as freedom from recurrence in those without measurable disease and complete clinical regression in those with measurable disease. Morbidity is assessed in terms of function and cosmesis. Survival is given from the time of the implant. RESULTS: There was local control in 13 patients maintained from 11 to 227 months (median, 39 months). There were two recurrences at 8 and 13 months in the first patient who received 17.3 Gy. Late complications consisted of mild eyelid telangiectasia, mild eyelid atrophy, and loss of eyelashes in most patients. Five patients, four of whom had upper eyelid tumors, experienced a dry eye, which was managed with tear supplements, and one of these patients developed a cataract. A corneal ulcer developed in one eye, which later perforated after treatment of a subsequent bulbar melanoma and was exenterated. Another eye was enucleated after treatment for a subsequent melanoma in the fornix. Cosmesis was acceptable to patient and doctor in the other 12 patients. Vision was maintained in seven patients, reduced in two, and not recorded in the remaining three patients. Three patients died of hematogenous metastases at 44, 62, and 79 months after implant, one of bronchial carcinoma at 46 months and one of an astrocytoma at 39 months. All patients were clear of local disease. The remainder survived for a median of 45 months (range, 18-227 months). CONCLUSIONS: Iodine-125 brachytherapy can be used as an alternative to wide excision or exenteration of these tumors. There was good local control, reasonable maintenance of vision, and good cosmesis. PMID- 10811090 TI - A comparative study of two procedures for repair of involutional lower lid entropion. AB - OBJECTIVE: The study compares the use of Jones' retractor plication and the Wies procedure as a primary procedure for the repair of involutional lower lid entropion in the absence of horizontal lid shortening. DESIGN: Retrospective case series comparison. PARTICIPANTS: One hundred two cases with at least 6 months follow-up after primary surgery for involutional lower lid entropion performed between 1993 and 1996. INTERVENTIONS: Two groups were compared: Jones' retractor plication was performed in one group and the Wies procedure in the other; neither group had horizontal shortening of the lower eyelid. MAIN OUTCOME MEASURES: The rates of cure, recurrence, and overcorrection of the lower lid entropion. RESULTS: Jones' retractor plication was performed in 37 (36%) patients and the Wies procedure in 65 (64%). Two of the 37 (5%) Jones cases had recurrent entropion develop, both at 31 months after surgery, compared with 1 1 of 65 (17%) recurrences after the Wies procedure (P = 0.81); recurrence after Wies procedure was earlier, at a median of 6 months after surgery. There were 20 of 65 (31 %) overcorrections after the Wies procedure and 4 of 37 (11 %) after the Jones procedure (P < 0.02); one quarter of each group required secondary repair of the overcorrection (5 of 20 after failed Wies procedure; 25% after failed Jones procedure). Overall, there were 6 of 37 (16%) unsatisfactory results after the Jones procedure in contrast to 31 of 65 (48%) after the Wies procedure (P < 0.001). CONCLUSIONS: These data provide strong evidence (P < 0.001) that, in the absence of horizontal shortening of the lower eyelid, a successful outcome is more likely after Jones retractor plication than after the Wies procedure. PMID- 10811091 TI - Intraocular involvement in multifocal fibrosclerosis. AB - OBJECTIVE: To report the clinical and histopathologic findings of intraocular involvement in a patient with multifocal fibrosclerosis and the response of the patient's choroidal masses to external beam radiotherapy. DESIGN: Case report with clinicopathologic correlation of enucleated eyes. METHODS: The patient was studied by clinical observation, contact B- and A-scan ultrasonography, orbital magnetic resonance imaging, fluorescein angiography, indocyanine green angiography, choroidal biopsy, and gross and histopathologic examination of the enucleated eyes. Tissue obtained at an earlier laparotomy was also reviewed. External beam radiotherapy was used when high-dose corticosteroid and low-dose methotrexate therapy failed to decrease the size of the choroidal masses or improve the patient's vision. MAIN OUTCOME MEASURES: Changes in the clinical and ultrasonographic size of the choroidal masses, the clinical appearance of these masses, and the patient's visual acuity in response to external beam radiotherapy were monitored premortem. Histopathologic findings in the enucleated eyes were compared with the changes in previous abdominal and choroidal biopsy specimens and with tissue alterations reported in multifocal fibrosclerosis. RESULTS: Biopsy of the choroidal mass revealed a fibrosclerosing process similar to that found in the abdomen. The patient received external beam radiotherapy with disappearance of the masses. Fibrosclerosing changes similar to those seen in the abdomen were observed replacing the choroid in the enucleated eyes. CONCLUSIONS: Multifocal fibrosclerosis may involve the choroid with histopathologic changes similar to those that have been described in other locations in the body. External beam radiotherapy may be an effective treatment for intraocular involvement by multifocal fibrosclerosis. PMID- 10811092 TI - Efficacy and safety of cyclosporin A ophthalmic emulsion in the treatment of moderate-to-severe dry eye disease: a dose-ranging, randomized trial. The Cyclosporin A Phase 2 Study Group. AB - OBJECTIVE: To investigate the efficacy, safety, formulation tolerability, and optimal dosing of a novel cyclosporin A oil-in-water emulsion formulation for the treatment of moderate-to-severe dry eye disease. DESIGN: Randomized, multicenter, double-masked, parallel-group, dose-response controlled trial. PARTICIPANTS: Total enrollment: 162 patients; cyclosporin A groups: 129 patients; vehicle group: 33 patients. INTERVENTION: Patients instilled study medication (cyclosporin A ophthalmic emulsion 0.05%, 0.1%, 0.2%, or 0.4%, or vehicle) twice daily into both eyes for 12 weeks, followed by a 4-week posttreatment observation period. MAIN OUTCOME MEASURES: EFFICACY: rose bengal staining, superficial punctate keratitis, Schirmer tear test, symptoms of ocular discomfort, and the Ocular Surface Disease Index (OSDI; a measure of symptom frequency and impact on vision-related functioning). SAFETY: biomicroscopy, cyclosporin A blood levels, conjunctival microbiology, intraocular pressure, visual acuity, and monitoring of adverse events. RESULTS: In a subset of 90 patients with moderate-to-severe keratoconjunctivitis sicca, the most significant improvements with cyclosporin A treatment were in rose bengal staining, superficial punctate keratitis, sandy or gritty feeling, dryness, and itching, with improvements persisting into the posttreatment period in some treatment groups. There was also a decrease in OSDI scores, indicating a decrease in the effect of ocular symptoms on patients' daily lives. There was no clear dose-response relationship, but cyclosporin A 0.1% produced the most consistent improvement in objective and subjective end points and cyclosporin A 0.05% gave the most consistent improvement in patient symptoms. The vehicle also performed well, perhaps because of its long residence time on the ocular surface. There were no significant adverse effects, no microbial overgrowth, and no increased risk of ocular infection in any treatment group. The highest cyclosporin A blood concentration detected was 0.16 ng/ml. All treatments were well tolerated by patients. CONCLUSIONS: Cyclosporin A ophthalmic emulsions, 0.05%, 0.1%, 0.2%, and 0.4%, were safe and well tolerated, significantly improved the ocular signs and symptoms of moderate-to-severe dry eye disease, and decreased the effect of the disease on vision-related functioning. Cyclosporin A 0.05% and 0.1% were deemed the most appropriate formulations for future clinical studies because no additional benefits were observed with the higher concentrations. PMID- 10811093 TI - Amniotic membrane transplantation for ocular surface reconstruction in Stevens Johnson syndrome. AB - PURPOSE: To evaluate amniotic membrane transplantation (AMT) for ocular surface reconstruction in Stevens-Johnson syndrome (SJS). DESIGN: Prospective interventional case series. PARTICIPANTS: Ten consecutive patients (10 eyes) with SJS that underwent AMT as the first step in staged ocular surface reconstruction were included. METHODS: Amniotic membrane was processed under sterile conditions from a fresh placenta obtained from cesarean section in a seronegative pregnant woman and stored at -700 degrees C. Symblepharon release, excision of epibulbar fibrous tissue, and clearing of the fibrovascular membrane over the cornea was performed in all cases. Amniotic membrane covered the entire bulbar surface up to the fornices in five eyes; cornea and the perilimbal area in two eyes; cornea, the inferior bulbar surface, and the lower fornix in two eyes; and cornea and the superior bulbar surface in one eye. Obliterated fornices were deepened by use of fornix-formation sutures in all eyes. Symblepharon ring was placed postoperatively for 3 weeks to 2 months. Mean postoperative follow-up was 13.5 months (SD, +3.8 months; range, 9-30 months). MAIN OUTCOME MEASURES: Restoration of adequate bulbar surface free of symblepharon and good fornix depth were the main outcome measures. RESULTS: Complete corneal reepithelization occurred in all eyes between 1 and 6 weeks. Adequate bulbar surface and fornix depth were achieved in nine eyes, all of which were free of symblepharon at the final follow up visit. Cicatricial entropion resolved in four of five lower eyelids and one of two upper eyelids after AMT. One patient had a central corneal melt that required or necessitated a penetrating keratoplasty. CONCLUSIONS: AMT restores adequate bulbar surface and fornix depth and prevents recurrence of symblepharon in severe cases of SJS. PMID- 10811094 TI - Amniotic membrane transplantation for acute chemical or thermal burns. AB - PURPOSE: To determine whether preserved human amniotic membrane (AM) can be used to treat ocular burns in the acute stage. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Thirteen eyes from 11 patients with acute burns, 10 eyes with chemical burns and 3 with thermal burns of grades II III (7 eyes) and grade IV (6 eyes), treated at 7 different facilities. METHODS: Patients received amniotic membrane transplantation (AMT) within 2 weeks after the injury. MAIN OUTCOME MEASURES: Integrity of ocular surface epithelium and visual acuity during 9 months of follow-up. RESULTS: Ten patients were male and one patient was female; most were young (38.2 +/- 10.6 years). For a follow-up of 8.8 + 4.7 months, 11 of 13 eyes (84.63%) showed epithelialization within 2 to 5 weeks (23.7 +/- 9.8 days), and final visual acuity improved > or = 6 lines (6 eyes), 4 to 5 lines (2 eyes), and 1 to 3 lines (2 eyes); only one eye experienced a symblepharon. Eyes with burns of grade II to III showed more visual improvement (7.3 +/- 3 lines) than those with burns of grade IV (2.3 +/- 3.0 lines; P < 0.05, unpaired t test). In the group with grade II or III burns, none had limbal stem cell deficiency. All eyes in the group with grade IV burns did experience limbal stem cell deficiency. CONCLUSIONS: Amniotic membrane transplantation is effective in promoting re-epithelialization and reducing inflammation, thus preventing scarring sequelae in the late stage. In mild to moderate burns, AMT alone rapidly restores both corneal and conjunctival surfaces. In severe burns, however, it restores the conjunctival ocular surface without debilitating symblepharon and reduces limbal stromal inflammation, but does not prevent limbal stem cell deficiency, which requires further limbal stem cell transplantation. These results underscore the importance of immediate intervention in the acute stage of eyes with severely damaged ocular surface. Further prospective randomized studies including a control group are required to determine the effectiveness of AMT in acute chemical and thermal burns of the eye. PMID- 10811095 TI - Goldmann perimetry in acromegaly: a survey of 307 cases from 1951 through 1996. AB - OBJECTIVE: Because visual pathway lesions are a common complication of pituitary tumors, visual field examinations in patients with acromegaly were studied. Proportion and outcome of visual field defects in patients with acromegaly were evaluated. DESIGN: Large, retrospective case series. PARTICIPANTS: We reviewed 307 cases of acromegaly seen from 1951 through 1996 at a single referral center. METHODS: Kinetic visual field testing had been performed with the Goldmann perimeter, and the frequency of visual field defects and their correlation with other clinical manifestations and characteristics of the adenoma were examined. MAIN OUTCOME MEASURES: Repeat visual field examinations. RESULTS: Of the 307 patients included in the analysis, a visual field defect that could be attributed to the pituitary adenoma was observed in 62 (20.2%) during follow-up. Visual field defects were bilateral in 38 (61.3%) of these cases. Patients with visual field abnormalities were significantly younger (P = 0.04), had larger tumors (P < 0.001), had more suprasellar extensions (P < 0.001), and had higher levels of growth hormone in their serum (P = 0.04) than patients free of visual field defects. At the end of the follow-up period, visual field examination remained abnormal in 32 (10.4%). Return to a normal visual field examination after treatment was more frequently observed in patients who were less than 40 years of age at the time of diagnosis (P = 0.004). Secondary empty sella syndrome was the main cause of visual field defects after treatment. Abnormal visual field, either at the time of diagnosis or during follow-up, decreased from 27% of patients between 1951 and 1975 to 15.4% of patients between 1976 and 1996, when modern neuroimaging techniques became available. CONCLUSIONS: Endocrinologic and neuroimaging follow-up of patients with acromegaly should be accompanied by ophthalmic assessment. Factors predictive of visual field defects have been identified. PMID- 10811097 TI - Combined exfoliation and pigment dispersion: paradigm of an overlap syndrome. AB - OBJECTIVE: To describe a series of patients with combined pigment dispersion syndrome (PDS) and exfoliation syndrome (XFS) and to introduce a concept, the overlap syndrome, to aid in assessing multiple risk factors for glaucomatous damage. DESIGN: Clinic-based, cross-sectional study. SETTING: New York Eye and Ear Infirmary. PARTICIPANTS: Twenty-six patients identified from the glaucoma database as having combined pigment dispersion syndrome-glaucoma and exfoliation syndrome-glaucoma. MAIN OUTCOME MEASURES: Quantification of patients with both pigment dispersion syndrome-glaucoma (PDS/PG) and exfoliation syndrome-glaucoma (XFS/XFG) and its clinical implications. RESULTS: Among the 26 patients (all white) having both XFS/XFG and PDS/PG, the average age was 64.3 +/- 9.8 years and 19 of 26 were men. All patients had bilateral PDS/PG. Bilateral XFS/XFG was present in 9 of 26 patients and, of the 17 patients with unilateral involvement, the left eye was affected in 13. CONCLUSIONS: Both XFS and PDS are common. Middle aged patients with known PDS/PG should be suspected of having the onset of XFS if one eye escapes intraocular pressure control. Patients with unilateral XFG at presentation may also have signs of PDS/PG, often remitted. We define the term overlap syndrome to describe the sequential appearance over time of two or more risk factors for glaucomatous damage. The appearance of a new risk factor in a patient whose condition has been stable can alter the course and prognosis of the disease. This concept should prove useful in dealing with secondary and normal tension glaucomas. PMID- 10811096 TI - Epidemiology of angle-closure glaucoma: prevalence, clinical types, and association with peripheral anterior chamber depth in the Egna-Neumarket Glaucoma Study. AB - OBJECTIVE: To assess the prevalence of primary angle-closure glaucoma (PACG), the frequency of its different clinical presentations, and its association with peripheral anterior chamber depth in a defined population in Northern Italy. DESIGN: Cross-sectional epidemiologic study in a defined population. PARTICIPANTS: All subjects resident in the Egna-Neumarkt area of the South Tyrol Region (Northern Italy) and more than 40 years of age were invited to undergo an ophthalmologic examination. INTERVENTION: After the screening examination, subjects with suspected glaucoma were re-examined at the screening center to confirm the diagnosis. All cases that still proved suspect after the second examination underwent a third phase of investigations and were classified as healthy or as definitely glaucomatous. Each subject was examined according to a standard protocol, including medical history interview, refraction and visual acuity determination, ocular biomicroscopy, evaluation of peripheral anterior chamber depth by means of the Van Herick method, applanation tonometry, optic disc evaluation, and computerized perimetry. Gonioscopy was not performed during initial screening but only in all selected patients in the second and third phases of investigations. The diagnosis of PACG was made on the basis of the concomitant presence of at least two of the following criteria: intraocular pressure > or = 22 mmHg, glaucomatous optic disc abnormalities, glaucomatous visual field defects. In addition, biomicroscopic or gonioscopic evidence of angle closure was also necessary. MAIN OUTCOME MEASURES: Percentage distribution of peripheral anterior chamber depths, prevalence of angle-closure glaucoma, and frequency of the different PACG clinical presentations. RESULTS: Four thousand two hundred ninety-seven subjects were examined (73.9% overall participation rate). The peripheral depth of the anterior chamber according to the Van Herick method was grade 2 in 14.7%, grade 1 in 2.5%, and grade 0 in 0.3% of the population. The overall prevalence of angle-closure glaucoma was 0.6% (26 cases). Five of these were cases of previous acute attacks resolved by therapy, three were cases of chronic angle-closure after acute attacks, three were intermittent angle-closure glaucomas, and 15 were chronic angle-closure cases. CONCLUSIONS: Occludable angles were more frequent than in other white populations previously studied. The prevalence of PACG is not as low as is usually believed; this type of glaucoma accounts for more than a quarter of all glaucomas found in the Egna Neumarkt population. The most frequent clinical presentation is chronic angle closure glaucoma. PMID- 10811099 TI - Macular translocation. American Academy of Ophthalmology. AB - OBJECTIVE: This document describes macular translocation surgery and examines the evidence to answer four key questions about whether the surgery is effective in treating visual loss from age-related macular degeneration, the technique that is most effective, the complications that result from the procedure, and the relationship of the volume of surgery to the number of complications. METHODS: A literature search that was conducted in September 1999 retrieved 21 relevant citations, and the reference lists of these articles were consulted for additional citations. Panel members reviewed this information, and a methodologist reviewed and rated all articles as good, fair, or poor in quality. RESULTS: The published literature contains only information from case series, which lack a control group and randomization. They describe a number of approaches, all of which are evolving. Follow-up is limited and visual outcomes vary widely. Serious intraoperative and postoperative complications are frequent, with accompanying risk of visual loss. Surgical morbidity is also an important factor. CONCLUSIONS: To date no strong evidence exists with which to answer questions about the effectiveness of macular translocation surgery in treating visual loss from age-related macular degeneration or about the most effective technique. Published case series have reported a significant rate of complications. There is no evidence to date to indicate that the complication rate is related to the volume of surgery done. Randomized clinical trials are needed to determine whether macular translocation is a safe and effective treatment for visual loss from age-related macular degeneration. PMID- 10811098 TI - Ophthalmoscopic detectability of the parafoveal annular reflex in the evaluation of the optic nerve: an experimental study in rhesus monkeys. AB - OBJECTIVE: To evaluate whether assessment of the ophthalmoscopic visibility of the parafoveal annular reflex is helpful in the detection of optic nerve fiber and optic nerve damage. DESIGN: Experimental animal study. ANIMALS: The study was performed in rhesus monkeys, divided into four study groups, with no significant difference in ages of the animals in the various groups. Three of these groups had (1) experimental chronic high-pressure glaucoma (n = 37 monkeys), (2) experimental temporary occlusion of the central retinal artery (n = 19 monkeys), and (3) systemic atherosclerosis and arterial hypertension (n = 13 monkeys). The fourth group was the normal control group (n = 10 monkeys). METHODS: In 60-degree color fundus photographs, taken at baseline and at the end of the study, the ophthalmoscopic detectability of the parafoveal annular reflex was graded, the visibility of the retinal nerve fiber layer was assessed, and neuroretinal rim and parapapillary atrophy were measured. MAIN OUTCOME MEASURES: Ophthalmoscopic detectability of the parafoveal annular reflex; visibility of the retinal nerve fiber layer; neuroretinal rim area; size of parapapillary atrophy. RESULTS: The parafoveal annular reflex was significantly (P < 0.0001) better detectable at baseline of the study than at the end of the study. In all study groups, detectability of the parafoveal annular reflex decreased significantly (P < 0.0001) with decreasing visibility of the retinal nerve fiber layer. In the glaucoma group, parafoveal annular reflex detectability additionally decreased significantly (P < 0.001) with increasing area of beta zone of parapapillary atrophy and with decreasing neuroretinal rim area. In the normal group, the parafoveal annular reflex detectability decreased with increasing age, parallel to a loss in the visibility of the retinal nerve fiber layer. In 23 of 25 (92%) eyes with a complete loss of the visibility of the retinal nerve fiber layer at the end of the study, the parafoveal annular reflex was no longer detectable. CONCLUSIONS: The findings suggest that evaluation of the detectability of the parafoveal annular reflex in routine ophthalmoscopy is a useful additional tool in the qualitative assessment of the retinal nerve fiber layer and optic nerve. PMID- 10811100 TI - Ligand responsiveness in human prostate cancer: structural analysis of mutant androgen receptors from LNCaP and CWR22 tumors. AB - Androgen receptors (ARs) belong to the family of hormone receptors that are ligand-dependent transcription factors. Endocrine therapy provides effective treatment for prostate cancer until mutations arise that alter the ligand responsiveness of AR. In this study, structural models were developed for the functional domains of human AR by homology modeling from crystal structures of closely related nuclear receptors. These models were used to locate the sites of two frequently occurring mutations in prostate cancer. The substitutions that develop in LNCaP (threonine-->alanine at residue 877) and CWR22 (histidine- >tyrosine at residue 874) tumor cell lines are both located on helix 11 that forms part of the ligand-binding pocket. However, the results suggest that these mutations influence ligand responsiveness by completely different mechanisms. Residue 877 contacts the ligand directly, and substitution at this site alters the stereochemistry of the binding pocket. Thus, the LNCaP mutation apparently broadens the specificity of ligand recognition. In contrast, residue 874 is located down the helical axis, projects away from the ligand pocket, and does not contact ligand. The side chain of residue 874 lies in a cavity between helices 11 and 12. Substitution of tyrosine for histidine 874 in CWR22 tumors may affect a conformational change of helix 12 and, thus, influence binding of coactivator proteins and their regulatory effect on transcriptional activation. PMID- 10811101 TI - Inducible chemoresistance to 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothe cin (CPT-11) in colorectal cancer cells and a xenograft model is overcome by inhibition of nuclear factor-kappaB activation. AB - Limited studies have indicated that some chemotherapy agents activate the transcription factor nuclear factor-kappaB (NF-kappaB), and that this leads to suppression of the apoptotic potential of the chemotherapy. In contrast, it was reported recently that stable inhibition of NF-kappaB in four different cancer cell lines did not lead to augmentation of the chemotherapy-induced apoptosis. In this study, we have focused on colorectal cancer, which is known to be highly resistant to genotoxic chemotherapy and gamma irradiation. We show that the topoisomerase I inhibitor 7-ethyl-10-[4-(1-piperidino)-1 piperidino]carbonyloxycamptothecin (CPT-11) activates NF-kappaB in most colorectal cancer cell lines. We then examine a therapeutic strategy that uses adenovirus-mediated transfer of the super-repressor IkappaBalpha to inhibit NF kappaB activation as an adjuvant approach to promote chemosensitivity in colorectal tumor cells to treatment with CPT-11. These data demonstrate that the protection from apoptosis induced in response to CPT-11 treatment is effectively inhibited by the transient inhibition of NF-kappaB in a variety of human colon cancer cell lines and in a tumor xenograft model, resulting in a significantly enhanced tumoricidal response to CPT-11 via increased induction of apoptosis. These findings indicate that the activation of NF-kappaB by chemotherapy is an important underlying mechanism of inducible chemoresistance. PMID- 10811102 TI - Nijmegen breakage syndrome disease protein and MRE11 at PML nuclear bodies and meiotic telomeres. AB - Nijmegen breakage syndrome is a disease characterized by immunodeficiency, genomic instability, and cancer susceptibility. The gene product defective in Nijmegen breakage syndrome, p95, associates with two other proteins, MRE11 and RAD50. Here we demonstrate that in the absence of DNA damage, a portion of p95 and MRE11 is concentrated in PML nuclear bodies (NBs); MRE11 localization to the NBs is p95-dependent. In mammalian meiocytes, these proteins are specifically found at the telomeres. These results implicate the NBs in the maintenance of genomic stability and suggest that p95 and MRE11 may have roles in telomere maintenance in mammals, analogous to the role their homologues play in yeast. PMID- 10811103 TI - Molecular anatomy of BCL6 translocations revealed by long-distance polymerase chain reaction-based assays. AB - BCL6 translocations involve not only immunoglobulin (IG) genes but also a number of non-IG loci as partners. Junctional sequences of three IG/BCL6 translocations were readily obtained by long-distance PCR. In cases where partner loci were not determined, we developed a long-distance inverse PCR method, which amplifies unknown fragments flanked by known BCL6 sequences. Using these two long-distance PCR-based approaches, we cloned junctional areas of BCL6 translocations from a total of 58 cases of B-cell tumors. Nucleotide sequencing and database searches revealed that 30 cases involved IGs as partners: IG heavy chain gene in 22, IG kappa light chain gene in 1, and IG lambda light chain gene in 7. In contrast, 23 cases affected non-IG loci, including the H4 histone gene, heat shock protein genes HSP89alpha and HSP90beta, and PIM-1 proto-oncogene. On der(3) chromosomes, complete sets of the promoters of these partner genes replaced that of BCL6 in the same transcriptional orientation. These results suggest that BCL6 gene affected by the translocation is transcriptionally activated by a variety of stimuli, including cell cycle control, changes in the physical environment, and response to cytokines. Break points on BCL6 occurred within the major translocation cluster, and we identified a 120-bp hyper-cluster region a short distance from the 3' end of exon 1. Gel mobility-shift assay suggested the presence of a protein(s) that bound to this particular region. PMID- 10811104 TI - The Fhit tumor suppressor protein regulates the intracellular concentration of diadenosine triphosphate but not diadenosine tetraphosphate. AB - To determine the role of the FHIT tumor suppressor gene product, a diadenosine 5',5'''-P1,P3-triphosphate (Ap3A) hydrolase, in the regulation of the concentration of Ap3A and diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) in vivo, the levels of the adenosine(5')triphospho(5')nucleoside (Ap3N) and adenosine(5')tetraphospho(5')nucleoside (Ap4N) families were measured by luminometry in a number of human cell lines and correlated with the expression of Fhit determined by immunoblotting. Fhit-positive cells had no Ap3N or a very low level of Ap3N, whereas most Fhit-negative cells had Ap3N in the range 0.2-0.9 pmol/10(6) cells. Ap4N (mean value, 0.17 pmol/10(6) cells) did not correlate with Fhit expression. The results suggest that Fhit efficiently metabolizes Ap3A and Ap3N but not Ap4A or Ap4N in vivo. PMID- 10811105 TI - Specific c-kit mutations in sinonasal natural killer/T-cell lymphoma in China and Japan. AB - Sinonasal lymphoma is one of the constituents of lethal midline granuloma, which is a clinical term for progressive, destructive lesions affecting the midline of the face. The majority of sinonasal lymphomas, especially those showing polymorphous patterns of proliferation and thus termed polymorphic reticulosis, recently were categorized as sinonasal natural killer/T-cell lymphomas. They are more prevalent in Asia than Europe or North America and are associated with EBV infection. Twenty-three cases with sinonasal natural killer/T-cell lymphomas were collected from two high-incidence regions: Beijing, China (14 cases) and Osaka, Japan (9 cases). c-kit mutations were analyzed on paraffin-embedded specimens by PCR-single-strand conformation polymorphism followed by direct sequencing; the c kit proto-oncogene encodes a receptor of tyrosine kinase, which plays an important role in the regulation of normal and neoplastic hematopoiesis by the interaction with its specific ligand, termed stem cell factor. Twelve single nucleotide substitution mutations were seen in 23 cases. Ten of 14 Chinese cases (71.4%) had mutations at exon 11 or exon 17, whereas only two of nine Japanese cases (22.2%) had mutations, showing a significant difference in frequency between Chinese and Japanese cases. Furthermore, seven of eight mutations (92%) in exon 17 occurred at codon 825 and three of four mutations (75%) in exon 11 occurred at codon 561. Such a specificity has not been reported before, and these results, taken together, suggest that location-specific differences in etiological factors cause specific mutations in c-kit gene. PMID- 10811106 TI - Progesterone receptor gene polymorphism is associated with decreased risk for breast cancer by age 50. AB - In a population-based case-control study for breast cancer before the age of 51 years, 554 cases and 559 age-matched controls were genotyped for the polymorphic progesterone receptor allele PROGINS. Breast cancer risk was decreased in women carrying the PROGINS allele. The odds ratio adjusted for age and study region was 0.76 [95% confidence interval (CI), 0.58-1.00]. Compared with wild-type A1/A1 homozygotes, the odds ratio for A1/A2 heterozygotes and A2/A2 homozygotes was 0.82 (95% CI, 0.62-1.08) and 0.27 (95% CI, 0.10-0.74), respectively, suggesting a gene dosage effect of the A2 allele. There was suggestive evidence for a differential effect by menopausal status (P = 0.07) and by family history of breast cancer (P = 0.15). PMID- 10811107 TI - Kinetics of plasma Epstein-Barr virus DNA during radiation therapy for nasopharyngeal carcinoma. AB - We studied the kinetics of circulating EBV DNA in the plasma of nasopharyngeal carcinoma (NPC) patients. Serial weekly sampling of 10 NPC patients revealed a rapid decline in plasma EBV DNA concentration after treatment. In two subjects, an initial rise in the circulating EBV DNA level was observed immediately after treatment initiation. Plasma EBV DNA levels were monitored daily during the first treatment week in a second cohort of five patients, and the results indicated that an initial rise in plasma EBV DNA concentration could be observed in all subjects during the first treatment week. This observation is consistent with the liberation of EBV DNA after therapy-induced cancer cell death. After this initial rise, plasma EBV DNA concentration was found to decay with a median half-life of 3.8 days (interquartile range, 2.4-4.4 days). Kinetic analysis of circulating tumor-derived DNA during treatment may be a powerful tool for evaluating the in vivo response of NPC and other tumors to antineoplastic treatment and may improve our understanding of the biology of plasma nucleic acids. PMID- 10811108 TI - Identification of differential methylation of the WT1 antisense regulatory region and relaxation of imprinting in Wilms' tumor. AB - Wilms' tumor (WT) is associated with loss of heterozygosity at chromosome 11p13, the site of the Wilms' tumor suppressor gene, WT1. Although the preferential loss of maternal alleles suggested that differential allelic expression of WT1 might occur, this has not been evident in normal fetal tissues or WTs. In this study, we show that the WT1 antisense regulatory region is differentially methylated, with Southern blot analysis of four loss of heterozygosity-negative WTs and their corresponding normal kidneys indicating that allelic methylation is lost in WTs. Reverse transcription-PCR expression analysis correlates methylation with monoallelic expression of the antisense WT1 transcript (WT1-AS) in normal kidney. However, WTs display hypomethylation and biallelic expression of WT1-AS. Our findings are consistent with imprinting of WT1-AS in normal kidney and the relaxation of imprinting in Wilms' tumorigenesis. This identifies the WT1 antisense regulatory region in intron 1 as a primary site for epigenetic deregulation at chromosome 11p13 in WTs. PMID- 10811109 TI - The Ras-mitogen-activated protein kinase pathway is critical for the activation of matrix metalloproteinase secretion and the invasiveness in v-crk-transformed 3Y1. AB - To search for the intracellular signaling pathway critical for the secretion of matrix metalloproteinases (MMP), we studied the effects of dominant negative Ras (S17N Ras) and dominant negative MEK1 (MEK1AA) expression in v-crk-transformed 3Y1. Expression of either S17N Ras or MEK1AA dramatically suppressed the augmented secretion of MMP-2 and MMP-9 in v-crk-transfected 3Y1. Similarly, a Ras farnesyltransferase inhibitor, manumycin A, and a MEK1 inhibitor, U0126, suppressed MMP secretion in a dose-dependent manner, whereas a PI3 kinase inhibitor, wortmannin, could not. In addition, the suppression of MMP secretion by S17N Ras showed good correlation with the inhibition of in vitro invasiveness of the cells. In contrast, expression of dominant negative C3G did not suppress MMP secretion, although it substantially blocked the c-Jun N-terminal kinase activation. Taken together, the Ras-MEK1 pathway, but not the C3G-JNK pathway, seems to play a key role in the activation of MMP secretion and, hence, the invasiveness of v-crk-transformed cells. PMID- 10811110 TI - Levels of E2F-1 expression are higher in lung metastasis of colon cancer as compared with hepatic metastasis and correlate with levels of thymidylate synthase. AB - We recently reported that forced overexpression of the transcription factor E2F-1 in human HT-1080 fibrosarcoma cells resulted in corresponding high levels of thymidylate synthase (TS) and resistance to 5-fluoropyrimidines (D. Banerjee et al., Cancer Res., 58: 4292-4296, 1998). Because colorectal metastasis to the lung has higher TS levels than liver metastasis and is less responsive to treatment with 5-fluorouracil (R. Gorlick et al., J. Clin. Oncol., 16: 1465-1469, 1998), it was, therefore, of interest to measure E2F-1 expression in these tumors. In contrast to marginally increased levels of dihydrofolate reductase and topoisomerase I in lung metastasis as compared with liver metastasis, lung tumors had a 5-fold increase in E2F-1 expression as compared with liver tumors, corresponding to the relative levels of TS in these metastases. These data indicate that there exists a close correlation between E2F-1 and TS levels and provide a rationale for targeting this transcription factor, ie., E2F-1, for the treatment of certain cancers. PMID- 10811111 TI - Inactivation of the DNA repair gene O6-methylguanine-DNA methyltransferase by promoter hypermethylation is associated with G to A mutations in K-ras in colorectal tumorigenesis. AB - O6-methylguanine DNA methyltransferase (MGMT) is a DNA repair protein that removes mutagenic and cytotoxic adducts from the O6 position of guanine. O6 methylguanine mispairs with thymine during replication, and if the adduct is not removed, this results in conversion from a guanine-cytosine pair to an adenine thymine pair. In vitro assays show that MGMT expression avoids G to A mutations and MGMT transgenic mice are protected against G to A transitions at ras genes. We have recently demonstrated that the MGMT gene is silenced by promoter methylation in many human tumors, including colorectal carcinomas. To study the relevance of defective MGMT function by aberrant methylation in relation to the presence of K-ras mutations, we studied 244 colorectal tumor samples for MGMT promoter hypermethylation and K-ras mutational status. Our results show a clear association between the inactivation of MGMT by promoter hypermethylation and the appearance of G to A mutations at K-ras: 71% (36 of 51) of the tumors displaying this particular type of mutation had abnormal MGMT methylation, whereas only 32% (12 of 37) of those with other K-ras mutations not involving G to A transitions and 35% (55 of 156) of the tumors without K-ras mutations demonstrated MGMT methylation (P = 0.002). In addition, MGMT loss associated with hypermethylation was observed in the small adenomas, including those that do not yet contain K-ras mutations. Hypermethylation of other genes such as p16INK4a and p14ARF was not associated with either MGMT hypermethylation or K-ras mutation. Our data suggest that epigenetic silencing of MGMT by promoter hypermethylation may lead to a particular genetic change in human cancer, specifically G to A transitions in the K-ras oncogene. PMID- 10811112 TI - WRN or telomerase constructs reverse 4-nitroquinoline 1-oxide sensitivity in transformed Werner syndrome fibroblasts. AB - WRN encodes a RecQ helicase, which is mutated in Werner syndrome. Werner syndrome is a genetic condition of young adults characterized by premature aging, limited replicative capacity of cells in vitro, and increased cancer risk. Telomerase is a reverse transcriptase that extends the G-rich strand of telomeric DNA. Primary cells in vitro typically lack telomerase activity and undergo senescence, whereas telomerase is reactivated in many, but not all, tumors. The roles of the two genes are not known to be related. Here we report the development of an effective colony-forming assay in which a SV40-transformed Werner fibroblast cell line is 6 18-fold more sensitive to 4-nitroquinoline 1-oxide than SV40-transformed normal cell lines. The sensitivity can be partially reversed by transfecting a normal WRN gene but not a mutated WRN gene into the cells. Curiously, the sensitivity can be reversed equally well by transfecting a telomerase gene (TERT) into the cells. These data indicate the possibility of an interdependent function of these two genes. PMID- 10811113 TI - Nitric oxide inhibits apoptosis via AP-1-dependent CD95L transactivation. AB - Several inducers of cytotoxic stress promote apoptotic cell death, which, at least in some cases, involves the CD95/CD95 ligand (CD95L) pathway. The induction of the CD95/CD95L pathway can be activated by the activator protein-1 (AP-1) mediated up-regulation of the CD95L promoter, which is responsible for the induction of apoptosis elicited by stimuli such as etoposide. We show that nitric oxide (NO) represents a regulatory element able to block apoptosis by interfering with this loop. Etoposide- and C6-ceramide-induced apoptosis in Jurkat T cells with different kinetics. Cell death was accompanied by an increase in DNA-binding activity of the transcription factor AP-1, transactivation of the AP-1 site containing CD95L promoter, and caspase 3-like protease activation. Using different NO-releasing compounds, we found that apoptosis was prevented in a dose dependent manner. Furthermore, in both models of apoptosis, NO-releasing compounds dose-dependently reduced: (a) the number of the titratable thiol groups (cysteine residues) of c-Jun; (b) induction of AP-1 DNA-binding activity; (c) AP 1-driven transactivation of the CD95L promoter; and (d) caspase activation. In conclusion, our data demonstrate that NO can modulate cell death at an upstream level, by interfering with the ability of AP-1 to induce CD95L expression. PMID- 10811114 TI - Tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in androgen-independent prostate cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been reported to induce cell death in a variety of transformed cells but spared the normal cells. In this study, we examined its potential against advanced prostate cancer cells. Treatment of PC-3 and DU145 cells with TRAIL caused a rapid apoptotic cell death, whereas tumor necrosis factor-alpha (TNF-alpha) is ineffective unless in the presence of the protein synthesis inhibitor cycloheximide. The induction of apoptosis by TRAIL in PC-3 cells was mediated by a death receptor, DR 4, and the downstream caspases. Treatment of PC-3 cells with TRAIL also activated c-Jun NH2 terminal kinase 1 (JNK1); however, inhibition of JNK1 activation by its dominant negative mutant had little effect on TRAIL-induced apoptosis. Furthermore, TRAIL weakly stimulated nuclear factor kappaB activity in PC-3 cells. Interestingly, activation of nuclear factor kappaB pathway by pretreatment with TNF-alpha did not prevent the induction of apoptosis by TRAIL. These data indicate that TRAIL triggers apoptosis in advanced prostate cancer cells through the activation of caspase cascades, which appears to be independent of TNF-alpha- and JNK-mediated mechanisms. PMID- 10811115 TI - Identification of the messenger RNA for human cutaneous fatty acid-binding protein as a metastasis inducer. AB - Using our recently developed systematic differential display and complete comparison of gene expression approaches combined with other methods, we have identified a large number of mRNAs that are expressed differentially between benign and malignant human cells. One such mRNA that is common to prostate and breast carcinoma cell lines encodes the human cutaneous fatty acid-binding protein (C-FABP). Northern and slot blot analyses confirm that the expression levels of C-FABP mRNA in the malignant prostate and breast carcinoma cell lines are 4.9+/-0.9- to 16.9+/-2.1-fold higher than those expressed in the benign cell lines. A similar difference between the benign and malignant cell lines was also detected at the protein level. In situ hybridization experiments have detected overexpression of the mRNA for C-FABP in human prostate carcinoma tissues. Transfection of a C-FABP expression construct into the benign, nonmetastatic rat mammary epithelial cell line Rama 37 and inoculation of the C-FABP expression transfectants into syngeneic Wistar-Furth rats produce a significant number (P < 0.05) of animals with metastases (6 of 26 animals), whereas the control transfectants generated by the vector alone yield no such metastases. Measurements of mRNA and protein levels with Northern and Western blotting show that C-FABP is not expressed in the control transfectant cells produced by the vector alone but is highly expressed in the pool of C-FABP transfectants and-the sublines established from their metastases. Immunocytochemical staining with antibodies to C-FABP shows that C-FABP is not expressed in the primary tumors developed from the control transfectants that have failed to metastasize, but it is expressed in both the primary tumors developed from the C-FABP transfectants and their metastases. Reinoculation of the sublines established from metastases in syngeneic rats has produced a higher proportion (50%) of animals (7 of 14 animals) with metastases than that obtained in the first-round inoculations, indicating that the metastatic clones have been preferentially selected from the original pool of metastatic and nonmetastatic transfectant clones. These results have demonstrated that elevated expression of C-FABP can induce metastasis and that metastatic capability has been transferred in a genetically dominated manner in this Rama 37 model. Thus, we suggest that C-FABP is a metastasis-inducing gene, and under suitable conditions, it may induce metastasis of some human cancers. PMID- 10811116 TI - Ursolic acid inhibits cyclooxygenase-2 transcription in human mammary epithelial cells. AB - We investigated the effects of ursolic acid, a chemopreventive agent, on the expression of cyclooxygenase-2 (COX-2) in phorbol 12-myristate 13-acetate (PMA) treated human mammary and oral epithelial cells. Treatment with ursolic acid suppressed PMA-mediated induction of COX-2 protein and synthesis of prostaglandin E2. Ursolic acid also suppressed the induction of COX-2 mRNA by PMA. Nuclear run offs revealed increased rates of COX-2 transcription after treatment with PMA, an effect that was inhibited by ursolic acid. Transient transfections indicated that the effects of PMA were mediated by a cyclic AMP response element in the COX-2 promoter. Ursolic acid inhibited PMA-mediated activation of protein kinase C, extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinases. Treatment with PMA increased activator protein 1 activity and the binding of c-Jun to the cyclic AMP response element of the COX 2 promoter, effects that were blocked by ursolic acid. These data are important for understanding the anticancer and anti-inflammatory properties of ursolic acid. PMID- 10811117 TI - Increased gene expression of brown fat uncoupling protein (UCP)1 and skeletal muscle UCP2 and UCP3 in MAC16-induced cancer cachexia. AB - Weight loss in cancer cachexia is attributable to decreased food intake and/or enhanced energy expenditure. We investigated the roles of the uncoupling proteins (UCPs) UCPI, -2, and -3 in a murine model of cachexia, the MAC16 adenocarcinoma. Weight fell to 24% below that of non-tumor-bearing controls (P < 0.01) 18 days after MAC16 inoculation, with significant reductions in fat-pad mass (-67%; P < 0.01) and muscle mass (-20%; P < 0.01). Food intake was 26-60% lower (P < 0.01) than in controls on days 17-18. Non-tumor-bearing mice, pair-fed to match MAC16 induced hypophagia, showed less weight loss (10% below controls, P < 0.01; 16% above MAC-16, P < 0.01) and smaller decreases in fat-pad mass (21% below controls, P < 0.01). Core temperature in MAC16 mice was significantly lower (-2.4 degrees C, P < 0.01) than in controls, and pair-feeding had no effect. MAC16 mice showed significantly higher UCP1 mRNA levels in brown adipose tissue (BAT) than in controls (+63%, P < 0.01), and pair-feeding had no effect. UCP2 and -3 expression in BAT did not differ significantly between groups. By contrast, UCP2 mRNA levels in skeletal muscle were comparably increased in both MAC16 and pair fed groups (respectively, 183 and 163% above controls; both, P < 0.05), with no significant difference between these two groups. Similarly, UCP3 mRNA was significantly higher than controls in both MAC16 (+163%, P < 0.05) and pair-fed (+253%, P < 0.01) groups, with no significant difference between the two experimental groups. Overexpression of UCP1 in BAT in MAC16-bearing mice may be an adaptive response to hypothermia, which is apparently induced by tumor products; increased thermogenesis in BAT could increase total energy expenditure and, thus, contribute to tissue wasting. Increased UCP2 and -3 expression in muscle are both attributable to reduced food intake and may be involved in lipid utilization during lipolysis in MAC16-induced cachexia. PMID- 10811118 TI - Functional assay for BRCA1: mutagenesis of the COOH-terminal region reveals critical residues for transcription activation. AB - The breast and ovarian cancer susceptibility gene product BRCA1 is a tumor suppressor, but its precise biochemical function remains unknown. The BRCA1 COOH terminus acts as a transcription activation domain, and germ-line cancer- predisposing mutations in this region abolish transcription activation, whereas benign polymorphisms do not. These results raise the possibility that loss of transcription activation by BRCA1 is crucial for oncogenesis. Therefore, identification of residues involved in transcription activation by BRCA1 will help understand why particular germ-line missense mutations are deleterious and may provide more reliable presymptomatic risk assessment. The BRCA1 COOH terminus (amino acids 1560-1863) consists of two BRCTs preceded by a region likely to be nonglobular. We combined site-directed and random mutagenesis, followed by a functional transcription assay in yeast: (a) error-prone PCR-induced random mutagenesis generated eight unique missense mutations causing loss of function, six of which targeted hydrophobic residues conserved in canine, mouse, rat, and human BRCA1; (b) random insertion of a variable pentapeptide cassette generated 21 insertion mutants. All pentapeptide insertions NH2-terminal to the BRCTs retained wild-type activity, whereas insertions in the BRCTs were, with few exceptions, deleterious; and (c) site-directed mutagenesis was used to characterize five known germ-line mutations and to perform deletion analysis of the COOH terminus. Deletion analysis revealed that the integrity of the most COOH terminal hydrophobic cluster (I1855, L1854, and Y1853) is necessary for activity. We conclude that the integrity of the BRCT domains is crucial for transcription activation and that hydrophobic residues may be important for BRCT function. Therefore, the yeast-based assay for transcription activation can be used successfully to provide tools for structure-function analysis of BRCA1 and may form the basis of a BRCA1 functional assay. PMID- 10811119 TI - Flavopiridol binds to duplex DNA. AB - Flavopiridol, the first potent cyclin-dependent kinase inhibitor to enter clinical trials, was recently found to be cytotoxic to noncycling cells. The present studies were performed to examine the hypothesis that flavopiridol, like several other antineoplastic agents that kill noncycling cells, might also interact with DNA. Consistent with this possibility, treatment of A549 human lung cancer cells with clinically achievable concentrations of flavopiridol resulted in rapid elevations of the DNA damage-responsive protein p53. In further studies, the binding of flavopiridol to DNA was examined in vitro by four independent techniques. Absorption spectroscopy revealed that addition of DNA to aqueous flavopiridol solutions resulted in a red shift of the flavopiridol lambda(max) from 311 to 344 nm, demonstrating an isosbestic point typical of changes seen with DNA-binding compounds. Reverse-phase high-performance liquid chromatography demonstrated that flavopiridol binds to genomic DNA to a similar extent as ethidium bromide and Hoechst 33258. Nuclear magnetic resonance spectroscopy revealed that DNA caused extreme broadening of flavopiridol 1H nuclear magnetic resonance signals that could be reversed by addition of ethidium bromide or by DNA melting, suggesting that flavopiridol binds to (and likely intercalates into) duplex DNA. Equilibrium dialysis demonstrated that the equilibrium dissociation constant of the flavopiridol-DNA complex (5.4+/-3.4 x 10(-4) M) was in the same range observed for binding of the intercalators doxorubicin and pyrazoloacridine to DNA. Molecular modeling confirmed the feasibility of flavopiridol intercalation into DNA and analysis of the effects of flavopiridol in the National Cancer Institute tumor cell line panel using the COMPARE algorithm demonstrated that flavopiridol most closely resembles cytotoxic antineoplastic intercalators. Collectively, these data suggest that DNA might be a second target of flavopiridol, providing a potential explanation for the ability of this agent to kill noncycling cancer cells. PMID- 10811120 TI - Proteasome inhibition circumvents solid tumor resistance to topoisomerase II directed drugs. AB - Physiological cell conditions, such as glucose deprivation and hypoxia, play a role in developing drug resistance in solid tumors. These tumor-specific conditions cause decreased expression of DNA topoisomerase IIalpha (topo IIalpha), rendering cells resistant to topo II-targeted drugs, such as etoposide and doxorubicin. We show here that inhibition of proteasome attenuated drug resistance by inhibiting topo IIalpha depletion induced by glucose starvation and hypoxia. topo IIalpha restoration was seen only at the protein levels, indicating that the topo IIalpha protein depletion occurred through a proteasome-mediated degradation mechanism. The stress-induced etoposide resistance was effectively prevented in vitro by the proteasome inhibitor lactacystin in both intrinsically resistant and sensitive tumor cells (colon cancer HT-29 and ovarian cancer A2780 cells, respectively). Furthermore, lactacystin effectively enhanced the antitumor activity of etoposide in the refractory HT-29 xenograft. These results indicate that lactacystin could serve as a new therapeutic agent to circumvent resistance to topo II-targeted chemotherapy in solid tumors. PMID- 10811121 TI - Immunity to murine breast cancer cells modified to express MUC-1, a human breast cancer antigen, in transgenic mice tolerant to human MUC-1. AB - The high incidence of breast cancer in women and the severity of the disease have stimulated a need for improved and novel forms of therapy. The product of the MUC 1 gene has been identified as a breast cancer-associated antigen in breast cancer patients. The gene has been cloned and sequenced. Transgenic mice were prepared that express human mucin and are naturally tolerant to the molecule, providing a unique opportunity to investigate immunotherapeutic strategies in experimental animals that might eventually be applied to breast cancer patients. A cell line (410.4) derived from a mouse mammary adenocarcinoma that arose in a BALB/c mouse was transduced with a retroviral vector (R1-MUC1-pEMSVscribe) that encoded MUC-1. After confirmation of the expression of human mucin, the cells (E3) were further modified by transduction with retroviral vectors encoding interleukin (IL)-2, IL 4, IL-12, or IFN-gamma to evaluate the effect of cytokine-secretion on the immunogenic properties of the cells in the MUC-1 transgenic mice. The results indicated that modification of the breast cancer cells to secrete IL-12 reduced and at times eliminated the tumorigenic growth properties of the cells. Under similar circumstances, progressively growing tumors formed in MUC-1 transgenic mice that received injections of unmodified E3 cells or with E3 cells modified to secrete IL-2, IL-4, or IFN-gamma. Immunity to breast cancer developed in MUC-1 transgenic mice that had rejected IL-12-secreting E3 cells because the animals were resistant to challenge with (non-cytokine-secreting) E3 cells. In vitro analyses confirmed the presence of T cell-mediated cytotoxicity toward the breast cancer cells in MUC-1 transgenic mice immunized with the IL-12-secreting cells. Our data obtained in a unique animal model system point toward an analogous form of therapy for breast cancer patients. PMID- 10811122 TI - Combination immunotherapy of primary prostate cancer in a transgenic mouse model using CTLA-4 blockade. AB - We have previously shown that antibodies to CTLA-4, an inhibitory receptor on T cells, can be effective at inducing regression of transplantable murine tumors. In this study, we demonstrate that an effective immune response against primary prostate tumors in transgenic (TRAMP) mice can be elicited using a strategy that combines CTLA-4 blockade and an irradiated tumor cell vaccine. Treatment of TRAMP mice at 14 weeks of age resulted in a significant reduction in tumor incidence (15% versus control, 75%), as assessed 2 months after treatment. Histopathological analysis revealed that treated mice had a lower tumor grade with significant accumulation of inflammatory cells in interductal spaces when treated with anti-CTLA-4 and a granulocyte-macrophage colony-stimulating factor expressing vaccine. Vaccination of nontransgenic mice with this regimen resulted in marked prostatitis accompanied by destruction of epithelium, indicating that the immune response was, at least in part, directed against normal prostate antigens. These findings demonstrate that this combinatorial treatment can elicit a potent antiprostate response and suggest potential of this approach for treatment of prostate cancer. PMID- 10811123 TI - Cytokine gene therapy of gliomas: induction of reactive CD4+ T cells by interleukin-4-transfected 9L gliosarcoma is essential for protective immunity. AB - Tumor cells genetically modified to secrete cytokines stimulate potent immune responses against peripheral and central nervous system tumors; however, variable results on the efficacy of this strategy for therapeutic intervention against established intracranial neoplasia have been reported. We have found that vaccination with rat 9L gliosarcoma cells expressing interleukin 4 (9LmIL4) induced a specific, protective, immune response against rechallenge with parental 9L tumors. In naive rats, sham-transfected 9L (9Lneo) tumors and 9LmIL4 tumors grew at comparable rates for 12-14 days, and then 9LmIL4 tumors regressed. After regression of 9LmIL4 tumors, rats were resistant to rechallenge with parental 9L cells. To investigate the mechanism(s) responsible for 9LmIL4-induced immunity, the phenotype and function of tumor-infiltrating lymphocytes (TILs) in 9Lneo and 9LmIL4 tumors were compared. In flow cytometric analyses, it was determined that CD4+ T cells were the predominant cell type in both 9Lneo and 9LmIL4 tumors at day 10. However, at the onset of regression (day 14), 9LmIL4 tumors were infiltrated predominantly by CD8+ T cells. To investigate functional aspects of the anti-9L tumor responses, we assessed the capacity of 9LmIL4 TILs to mediate specific lytic function or production of cytokines. In response to parental 9L, TILs isolated from day 14 9LmIL4 tumors were demonstrated to produce substantially greater amounts of IFN-gamma than did TILs from 9Lneo tumors. Although freshly isolated TILs from 9LmIL4 or control tumors did not lyse 9L cells in 51Cr-release cytotoxicity assays, specific cytotoxicity was demonstrable using TILs from day 14 9LmIL4 or splenocytes from 9LmIL4-bearing rats after their restimulation for 5 days with parental 9L tumor cells in vitro. Antibody blocking studies demonstrated that cytokine production and lytic activity by TILs, or splenocytes from 9LmIL4-immunized rats, were mediated in a T-cell receptor dependent fashion. Because interleukin-4 also promotes humoral responses, quantity and isotype of immunoglobulins in sera from 9Lneo or 9LmIL4-immunized rats were compared. The amount of IgG1 antibodies was significantly increased in sera from 9LmIL4-immunized rats compared to sera from 9Lneo-bearing rats. Experiments using sublethally irradiated, naive rats adoptively transferred with splenocytes and/or sera from 9LmIL4-immunized or naive rats demonstrated that immune cells, with or without immune sera, protected recipients from challenge with parental 9L. Immune sera provided no protection when given with lymphocytes from naive rats, and it did not enhance protection against parental 9L when given in conjunction with lymphocytes for 9LmIL4-immunized rats. In additional adoptive transfer experiments, an essential role for CD4+ T cells in immunity was observed because their depletion from among splenocytes of 9LmIL4-immunized rats eliminated the protective effective against 9L, whereas depletion of CD8+ cells resulted in a more limited effect on protection against 9L. These data suggest that strategies for inducing systemic, long-term tumor-specific reactivity among CD4+ T cells will be critical for the development of immunotherapy of gliomas. PMID- 10811124 TI - Photorepair prevents ultraviolet-induced apoptosis in human cells expressing the marsupial photolyase gene. AB - Photolyase absorbs blue light and employs the energy to remove UV-induced DNA damage, cyclobutane pyrimidine dimers, or pyrimidine pyrimidone (6-4) lesions. These enzymes have been found in many living organisms ranging from bacteria to aplacental mammals, but their photoreactivation effect, such as survival increase of UV-irradiated cells by light-illumination, has not been identified in placental mammals, including humans. Therefore, we introduced a photolyase gene derived from the marsupial rat kangaroo, Potorous tridactylus, into HeLa cells and established the first human cell line capable of photorepairing UV-induced pyrimidine dimers. Several clones were found to increase cell survival after UV irradiation when illuminated by fluorescent light. The induction of apoptosis by UV irradiation was investigated in these photoreactivation-proficient cells. Several typical features of the programmed cell death, such as internucleosomal DNA degradation, presence of subdiploid cells, loss of membrane integrity, and chromosomal condensation, were found to be induced by UV in the HeLa cells, but they can be reduced by photorepair. This implicates that cyclobutane pyrimidine dimers cause UV-induced apoptosis in human cells. PMID- 10811125 TI - A role for the p38 mitogen-acitvated protein kinase pathway in the transcriptional activation of p53 on genotoxic stress by chemotherapeutic agents. AB - The tumor suppressor p53 plays a central role in sensing damaged DNA and orchestrating the consequent cellular responses. However, how DNA damage leads to the activation of p53 is still poorly understood. In this study, we have found that the p38 mitogen-activated protein kinase (MAPK) plays a key role in the activation of p53 by genotoxic stress when provoked by chemotherapeutic agents. Indeed, we found that blockade of p38 prevents stimulation of the transcriptional activity of p53 and that activation of the p38 pathway is sufficient to stimulate p53 function. Furthermore, we observed that p38 does not affect the accumulation of p53 in response to DNA damage or its nuclear localization. In contrast, we observed that p38 associates physically with p53, and we provide evidence that this MAPK phosphorylates the NH2-terminal transactivation domain of p53 in serine 33, thereby stimulating its functional activity. Moreover, inhibition of the p38 MAPK diminished the apoptotic fraction of cells exposed to chemotherapeutic agents and increased cell survival, thus suggesting a role for p38 activation in the apoptotic response to genotoxic stress when elicited by drugs used in cancer therapy. PMID- 10811126 TI - Progressive increases in de novo methylation of CpG islands in bladder cancer. AB - We conducted a quantitative analysis of the extent of de novo methylation of four CpG islands in human urinary transitional cell carcinomas of different stages and grades to determine how frequently these CpG islands became methylated in transition cell carcinomas during progression. The CpG islands included exon 5 of PAX6, exon 2 of p16, the 5' end of the deleted in bladder cancer gene, and the 5' end of transmembrane protein containing epidermal growth factor and follistatin domains. These sequences were not methylated in normal urothelial tissues; however, 48 of the 54 tumors examined (89%) showed methylation levels in excess of 20% for at least one of the markers. The number of markers concurrently methylated in individual tumors increased with the stage of the tumor, with several of the more aggressive invasive cancers showing hypermethylation of all four markers compared with the less aggressive invasive cancers. However, considerable methylation defects were present in superficial, preinvasive, papillary tumors. These data demonstrate that 89% of bladder cancers have increased methylation of CpG islands relative to their normal counterparts and suggest the occurrence of a hypermethylator phenotype in which multiple independent CpG islands become concurrently methylated in individual tumors in a process associated with tumor progression. PMID- 10811127 TI - Allelic loss at SMAD4 in polyps from juvenile polyposis patients and use of fluorescence in situ hybridization to demonstrate clonal origin of the epithelium. AB - Juvenile polyposis syndrome (JPS; Online Mendelian Inheritance in Man2 174900) is a rare Mendelian disorder in which individuals have typical hamartomatous polyps within the gastrointestinal tract. The stromal element of the polyps has classically been thought to be the proliferative component, although epithelial malignancies (largely gastrointestinal cancers) occur more frequently than expected in JPS patients. Germ-line mutations in SMAD4 (DPC4) account for about a third of JPS cases. It has been postulated that the apparent paradox of a stromal lesion predisposing to epithelial malignancy can be resolved by the "landscaper" effect: an abnormal stromal environment affects the development of adjacent epithelial cells, and the resulting regeneration of damaged epithelium leads to an increased risk of cancer. We have found allele loss at the SMAD4 locus on 18q in polyps from JPS individuals with a germ-line SMAD4 mutation, showing that SMAD4 is acting as a tumor suppressor gene in JPS polyps, as it does in sporadic cancers of the gastrointestinal tract. Interphase fluorescence in situ hybridization showed deletion of one copy of SMAD4 in the epithelial component of JPS polyps, but not in the inflammatory infiltrate. Fluorescence in situ hybridization also suggested that a single copy of SMAD4 was present in stromal fibroblasts of JPS polyps. Thus, biallelic inactivation of SMAD4 occurs in both the epithelium and some of the stromal cells in these lesions, suggesting a common clonal origin. Epithelial malignancies almost certainly develop in juvenile polyposis through direct malignant progression of the epithelial component of the hamartomas. SMAD4/DPC4 probably acts as a "gatekeeper" tumor suppressor in juvenile polyps, and there is no need to invoke a "landscaper hypothesis." PMID- 10811128 TI - Genome-wide screen for allelic imbalance in a mouse model for neuroblastoma. AB - We have used the rat tyrosine hydroxylase promotor to overexpress MYCN in the neural crest of transgenic mice, resulting in a mouse model for neuroblastoma. Using PCR analysis of microsatellite markers, we conducted a genome-wide analysis in tumors from these animals. Regions of chromosomes 1, 3, 10, 11, 14, and 18 were affected in 20-50% of tumors. Analysis of a subset of these tumors by comparative genomic hybridization was consistent with the microsatellite data. The changes on mouse chromosomes 1, 11, 14, and 18 were syntenic with corresponding regions of loss of heterozygosity in human neuroblastoma, suggesting that genes implicated in the mouse tumors may also play a role in the pathogenesis of the human disease. One-third of the mouse tumors shared abnormalities on chromosomes 1, 3, and 10, whereas the remainder of tumors did not show this combination. These data suggest that genetic mutations on chromosomes 1, 3, and 10 cooperate in the pathogenesis of neuroblastoma and that neuroblastoma in the mouse arises from at least two distinct genetic pathways, one of which is dependent on lesions in chromosomes 1, 3, and 10, the other of which is not. PMID- 10811129 TI - Microsatellite instability at selected tetranucleotide repeats is associated with p53 mutations in non-small cell lung cancer. AB - Microsatellite alterations are useful clonal markers for the early detection of cancer. An increase in microsatellite instability has been observed at certain tetranucleotide repeat markers (AAAGn) in lung, head and neck, and bladder cancer. However, the genetic mechanism underlying these elevated microsatellite alterations at selected tetranucleotide repeat (EMAST) tumors is still unknown. The p53 gene plays an important role in maintaining genome integrity by repairing damaged DNA. Therefore, we tested 88 non-small cell lung cancers with a panel of 13 microsatellite markers previously shown to exhibit frequent instability and also performed p53 sequence analysis in these tumors. Thirty-one of these 88 cancers (35%) demonstrated a novel allele [EMAST(+)] in > or =1 of these 13 microsatellite markers. p53 mutations were detected in 50 of 88 (57%) cancers and were significantly (P = 0.001) more common in EMAST(+) tumors (25 of 31; 81%) than in EMAST(-) tumors (25 of 57; 44%). Among squamous cell cancers, p53 mutations were detected significantly (P = 0.04) more frequently in EMAST(+) tumors (17 of 19; 89%) than in EMAST(-) tumors (10 of 18; 55%). Similarly, among primary adenocarcinomas, p53 mutations were present in 67% of the EMAST(+) tumors and in 35% of EMAST(-) adenocarcinomas. None of the 31 EMAST(+) tumors demonstrated high frequency microsatellite instability when examined with a reference panel of five mono- and dinucleotide markers. Primary lung cancers with microsatellite alterations at selected tetranucleotide repeats have a high frequency of p53 mutations and do not display a phenotype consistent with defects in mismatch repair. PMID- 10811130 TI - Genetic instability and hematologic disease risk in Werner syndrome patients and heterozygotes. AB - Werner syndrome (WRN) is an uncommon autosomal recessive disease in which progeroid features are associated with genetic instability and an elevated risk of neoplasia. We have used the glycophorin A (GPA) somatic cell mutation assay to analyze genetic instability in vivo in WRN patients and heterozygotes. GPA variant frequencies were determined for 11 WRN patients and for 10 heterozygous family members who collectively carry 10 different WRN mutations. Genetic instability as measured by GPA O/N allele loss variant frequency was significantly increased, and this increase was strongly age-dependent in WRN patients. GPA O/N allele loss variants were also significantly elevated in heterozygous family members, thus providing the first evidence for in vivo genetic instability in heterozygous carriers in an autosomal recessive genetic instability syndrome. Our results and comparable data on other human genetic instability syndromes allow an estimate of the level of genetic instability that increases the risk of human bone marrow dysfunction or neoplasia. PMID- 10811131 TI - Role of extracellular matrix assembly in interstitial transport in solid tumors. AB - The extracellular matrix (ECM) may contribute to the drug resistance of a solid tumor by preventing the penetration of therapeutic agents. We measured differences in interstitial resistance to macromolecule (IgG) motion in four tumor types and found an unexpected correspondence between transport resistance and the mechanical stiffness. The interstitial diffusion coefficient of IgG was measured in situ by fluorescence redistribution after photobleaching. Tissue elastic modulus and hydraulic conductivity were measured by confined compression of excised tissue. In apparent contradiction to an existing paradigm, these functional properties are correlated with total tissue content of collagen, not glycosaminoglycan. An extended collagen network was observed in the more penetration-resistant tumors. Collagenase treatment of the more penetration resistant tumors significantly increased the IgG interstitial diffusion rate. We conclude that collagen influences the tissue resistance to macromolecule transport, possibly by binding and stabilizing the glycosaminoglycan component of the ECM. These findings suggest a new method to screen tumors for potential resistance to macromolecule-based therapy. Moreover, collagen and collagen proteoglycan bonds are identified as potential targets of treatment to improve macromolecule delivery. PMID- 10811132 TI - A critical step in metastasis: in vivo analysis of intravasation at the primary tumor. AB - Detailed evaluation of all steps in tumor cell metastasis is critical for evaluating the cell mechanisms controlling metastasis. Using green fluorescent protein transfectants of metastatic (MTLn3) and nonmetastatic (MTC) cell lines derived from the rat mammary adenocarcinoma 13762 NF, we have measured tumor cell density in the blood, individual tumor cells in the lungs, and lung metastases. Correlation of blood burden with lung metastases indicates that entry into the circulation is a critical step for metastasis. To examine cell behavior during intravasation, we have used green fluorescent protein technology to view these cells in time lapse images within a single optical section using a confocal microscope. In vivo imaging of the primary tumors of MTLn3 and MTC cells indicates that both metastatic and nonmetastatic cells are motile and show protrusive activity. However, metastatic cells show greater orientation toward blood vessels and larger numbers of host cells within the primary tumor, whereas nonmetastatic cells fragment when interacting with vessels. These results demonstrate that a major difference in intravasation between metastatic and nonmetastatic cells is detected in the primary tumor and illustrate the value of a direct visualization of cell properties in vivo for dissection of the metastatic process. PMID- 10811133 TI - A spontaneous murine melanoma lung metastasis comprised of host x tumor hybrids. AB - Cells from a lung metastasis, arising from Cloudman S91 melanoma cells implanted s.c. in the tail of a BALB/c nu/nu mouse, were comprised chiefly of host x tumor hybrids. These lung metastasis cells showed: (a) 30-40% increased DNA content; (b) resistance to 10(-4) M hypoxanthine, 4 x 10(-7) M aminopterin, and 1.6 x 10( 5) M thymidine (HAT) + G418; and (c) the presence in genomic DNA of genes for both wt and albino tyrosinase, reflecting the DBA/2J (Cloudman S91) and BALB/c mouse genotypes, respectively. Individual clones of lung metastasis cells expressed enhanced pigmentation, motility, and responsiveness to MSH/IBMX, a behavior similar to that recently reported for artificially generated melanoma x macrophage fusion hybrids. These similarities suggested that the host fusion partner generating the lung metastasis hybrids might have been a macrophage, although formal proof for this was not possible. The results provide the first direct evidence that host x tumor hybridization could serve as an initiating mechanism for melanoma metastasis. PMID- 10811134 TI - Anti-angiogenic cues from vascular basement membrane collagen. AB - Vascular basement membrane is an important structural component of blood vessels and has been shown to interact with and modulate vascular endothelial behavior during angiogenesis. During the inductive phase of tumor angiogenesis, this membrane undergoes many degradative and structural changes and reorganizes to a native state around newly formed capillaries in the resolution phase. Such matrix changes are potentially associated with molecular modifications that include expression of matrix gene products coupled with conformational changes, which expose cryptic protein modules for interaction with the vascular endothelium. We speculate that these interactions provide important endogenous angiogenic and anti-angiogenic cues. In this report, we identify an important antiangiogenic vascular basement membrane-associated protein, the 26-kDa NC1 domain of the alpha1 chain of type IV collagen, termed arresten. Arresten was isolated from human placenta and produced as a recombinant molecule in Escherichia coli and 293 embryonic kidney cells. We demonstrate that arresten functions as an anti angiogenic molecule by inhibiting endothelial cell proliferation, migration, tube formation, and Matrigel neovascularization. Arresten inhibits the growth of two human xenograft tumors in nude mice and the development of tumor metastases. Additionally, we show that the anti-angiogenic activity of arresten is potentially mediated via mechanisms involving cell surface proteoglycans and the alpha1beta1 integrin on endothelial cells. Collectively, our results suggest that arresten is a potent inhibitor of angiogenesis with a potential for therapeutic use. PMID- 10811135 TI - Human myeloid and lymphoid malignancies in the non-obese diabetic/severe combined immunodeficiency mouse model: frequency of apoptotic cells in solid tumors and efficiency and speed of engraftment correlate with vascular endothelial growth factor production. AB - Recent studies have suggested that non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice transplanted with human hematological malignancies show higher levels of engraftment compared with other strains. We used this model to compare xenotransplantability of human leukemia and lymphoma cell lines and to investigate angiogenesis in hematopoietic malignancies. Ten of 12 evaluated cell lines were able to engraft NOD/SCID mice within 120 days. A strong correlation was observed between the amount of vascular endothelial growth factor (VEGF) produced in vitro by cultured cells and the efficiency of tumor engraftment (r = 0.808; P = 0.001), and an inverse correlation was found between VEGF production and the time of tumor engraftment (r = -0.792; P = 0.006) and between VEGF production and the frequency of apoptotic/dead cells in solid tumors (r = -0.892; P = 0.007). Moreover, VEGF production correlated with the frequency of endothelial (CD31+/CD34+) cells in solid tumors (r = 0.897; P = 0.001). Taken together with in vitro data presented here and indicating that the VEGF antagonist Flt-1/Fc chimera inhibits leukemia and lymphoma cell proliferation, our findings support a role for tumor-derived VEGF in leukemia and lymphoma progression. Furthermore, the present study confirms previous observations indicating that VEGF expression may play a crucial role in xenotransplantability of human solid malignancies in SCID mice. The NOD/SCID model is promising for future evaluations of antiangiogenic drugs, alone or in combination with established chemo- or immunotherapy regimens. PMID- 10811136 TI - Establishment and characterization of a human lung cancer cell line NCI-H460 LNM35 with consistent lymphogenous metastasis via both subcutaneous and orthotopic propagation. AB - Lymphogenous metastasis is a common feature of human lung cancers, but very little is known about the underlying mechanism. In the present study, in vivo selection was carried out to obtain a highly lymphogenous metastatic subline of a human large cell carcinoma of the lung, NCI-H460. The resulting subline, termed NCI-H460-LNM35 (LNM35), was shown to metastasize to regional lymph nodes with a 100% incidence not only as a result of orthotopic intrabronchial (i.b.) implantation, but also as a result of conventional s.c. implantation. LNM35 has a short latency period, allowing for the collection of experimental data within 28 days after i.b. inoculation and 45 days after s.c. inoculation. It was noted that orthotopically i.b.-propagated LNM35 closely mimicked the clinical manifestations of human lung cancer patients by infiltrating into lymphatic vessels and metastasizing to the mediastinal lymph nodes. The LNM35 cell line is, to the best of our knowledge, the first human lung cancer cell line to be reported as having lymphogenous metastatic properties, and the observed 100% incidence by s.c. inoculation gives LNM35 a significant advantage even over previously reported human cancer cell lines of other origins. Comparisons between LNM35 and its parental NCI-H460 cell lines were also made with regard to expression levels and/or activities of various molecules that are thought to play a part in the metastatic process. We show here that the expression of cyclooxygenase 2 is increased in LNM35 and that a specific cyclooxygenase 2 inhibitor, nimesulide, can inhibit the invasion of LNM35 in vitro through Matrigel containing basement membrane components. PMID- 10811137 TI - Temporal progression of metastasis in lung: cell survival, dormancy, and location dependence of metastatic inefficiency. AB - Cancer metastasis is an inefficient process. The steps in metastasis responsible for this inefficiency and how metastatic inefficiency can vary in different locations within an organ remain poorly understood. B16F10 cells were injected to target mouse lung, and at sequential times thereafter we quantified in lung the time course of: (a) overall cell survival and metastatic development; and (b) local cell survival and growth with respect to the lung surface and specific interior structures. We found high rates of initial survival of cells trapped in the lung circulation, extravasation into lung tissue, and subsequent survival of extravasated solitary cells (74% at day 3) before metastasis formation. However, at the time of initial replication of metastatic cells a major loss of cells occurred. Although only a small proportion of injected cells started to form metastases, most of these developed into macroscopic tumors. Solitary cells found at later times were dormant. Thus, overall metastatic inefficiency was largely due to postextravasation events affecting solitary cells. Regionally within the lung, cells and metastases were randomly distributed to day 4, but by day 10 preferential tumor growth was found along the lung surface and around arterial and venous vessels. Thus, trapping and early growth of injected cells was unaffected by location within the lung, whereas subsequent metastatic growth was enhanced in specific microenvironments. This study: (a) quantifies early temporal and spatial progression of metastasis in lung; (b) documents persistence of solitary dormant cells; and (c) shows that metastatic inefficiency depends on the initiation of growth in a subset of extravasated cells, whereas continued growth of metastases occurs preferentially in specific tissue environments. PMID- 10811138 TI - Acquisition of chemoresistant phenotype by overexpression of the antiapoptotic gene testosterone-repressed prostate message-2 in prostate cancer xenograft models. AB - Testosterone-repressed prostate message-2 (TRPM-2) expression is highly up regulated in normal and malignant prostate cells after androgen withdrawal. Although recent studies have suggested a protective role of TRPM-2 expression against apoptosis in several experimental models, the functional role of TRPM-2 in chemotherapy-induced apoptosis remains undefined. Here, we demonstrated that overexpression of TRPM-2 in human androgen-dependent LNCaP prostate cancer cells by stable transfection rendered them highly resistant to paclitaxel treatment than control LNCaP cells, with a 20-fold higher IC50 through the inhibition of apoptotic cell death. In mice bearing TRPM-2-overexpressing LNCaP tumors, tumor volume and serum prostate-specific antigen increased two to three times faster after castration and paclitaxel treatment compared with mice bearing control tumors. We then tested the efficacy of combined treatment with antisense TRPM-2 oligodeoxynucleotide (ODN) and paclitaxel in the mouse androgen-dependent Shionogi tumor model. Antisense TRPM-2 ODN treatment significantly enhanced paclitaxel chemosensitivity of Shionogi tumor cells in a dose-dependent manner, reducing the IC50 by 75%. Combined treatment of Shionogi cells with 500 nM antisense TRPM-2 ODN and 10 nM paclitaxel-induced apoptosis, either agent alone did not. Adjuvant administration of antisense TRPM-2 ODN and polymeric micellar paclitaxel after castration resulted in reduced TRPM-2 levels in vivo and a significant delay of emergence of androgen-independent recurrent Shionogi tumors compared with administration of either agent alone. Furthermore, combined treatment of mice bearing androgen-independent recurrent Shionogi tumors with antisense TRPM-2 ODN and micellar paclitaxel inhibited tumor growth compared with treatment with either agent alone. Collectively, these findings demonstrate that TRPM-2 overexpression helps confer a chemoresistant phenotype through inhibition of apoptosis, and that antisense TRPM-2 ODN may be useful in enhancing the effects of cytotoxic chemotherapy in hormone-refractory prostate cancer. PMID- 10811140 TI - Concurrent and independent genetic alterations in the stromal and epithelial cells of mammary carcinoma: implications for tumorigenesis. AB - The high frequency of loss of heterozygosity (LOH) in epithelial cells of mammary ductal carcinoma in situ (DCIS) and IDC is a well known phenomenon, whereas the genetic abnormalities in the mammary stroma and its influence on the epithelial component have not been sufficiently studied. Using the PCR, we examined DNA extracts from microdissected stromal and epithelial tissues of 11 breast samples containing DCIS, including five cases associated with IDC. In each case, the mesenchymal tissue consisting of normal-appearing stroma at a distance from DCIS and IDC or stroma close to either DCIS or IDC was manually microdissected. Epithelial cells from morphologically clear-cut normal ducts and lobules, DCIS, and IDC were also microdissected. Twelve polymorphic DNA markers were tested to identify possible genetic alterations in the mesenchymal and epithelial cells on chromosomes 2p, 3p, 11q, 16q, and 17q. Samples from bilateral reduction mammoplasty from 10 women without any clinical, radiological, or pathological abnormalities were also selected as a control (reduction mammoplasty group). Whereas most cases (8/11, 73%) displayed at least one identical LOH in both epithelial and mesenchymal components, LOH at several loci was noted exclusively in stromal cells. The most frequent genetic alterations in the mesenchymal cells were at chromosomes 17q24, 16q23.1-24.2, 3p14.2, and 11q21-23.2, in 87.5, 62, 60, and 45% of informative cases, respectively. The LOH frequency in the stroma close to cancer ranged from 10 to 66.5% for DCIS and from 20 to 75% of informative cases for IDC. Furthermore, 10 of the 12 polymorphic markers revealed LOH in the stroma at a distance, ranging from 11 to 57% of informative cases. None of the control cases (women without any breast disease) revealed LOH either in the epithelial or in the stromal components. Our findings strongly support the concept of stromal-epithelial interaction in the development and progression of mammary neoplasia. Furthermore, this study suggests that genetic alterations in the stromal cells may precede genotypic changes in the epithelial cells. At least in some cases, the mammary stroma in DCIS or IDC apparently represents a neoplastic interactive component rather than a reactive response to the carcinoma. The frequent allelic loss (LOH) in the mammary stroma, identified in our study, may explain some of the fibroblastic abnormalities previously observed in patients with breast carcinoma or a variety of cancer-associated hereditary diseases. We conclude that the mammary stroma may play a key role in inducing neoplastic transformation of epithelial cells, recapitulating its role in normal mammary duct development. PMID- 10811139 TI - Aspirin inhibits tumor cell invasiveness induced by Epstein-Barr virus latent membrane protein 1 through suppression of matrix metalloproteinase-9 expression. AB - Matrix metalloproteinases (MMPs) are thought to play crucial roles in tumor invasion and metastasis. Because we have shown that EBV latent membrane protein 1 (LMP1) enhances MMP-9 expression by activation of nuclear factor (NF)-kappaB and activator protein (AP)-1 (T. Yoshizaki, et al., Proc. Natl. Acad. Sci. USA, 95: 3621-3626, 1998), we therefore tested whether up-regulation of MMP-9 by LMP1 could be correlated with enhanced invasiveness of tumor cells in vitro. Whether aspirin and sodium salicylate could reduce invasiveness and whether LMP1 could enhance MMP-9 expression in tumors grown in nude mice were also tested. C33A cells stably expressing LMP1 had increased expression of MMP-9 and showed greater invasion through reconstituted basement membrane compared with vector-transfected C33A cells (P < 0.02). Treatment with aspirin or sodium salicylate inhibited invasiveness of the LMP1-expressing C33A cells (P < 0.03) and suppressed both the LMP1-induced MMP-9 expression in zymographic analyses and LMP1-induced MMP-9 promoter activity in CAT reporter assays (P < 0.01). Endogenous MMP-2 levels were unaffected by either drug. Both drugs repressed the CAT activity of the truncated MMP-9 promoter construct, which only contained a binding site for AP-1, to the basal level (P < 0.05). Moreover, EMSA indicated that the effects of the salicylates were through the inhibition of not only NF-kappaB but also AP-1 binding activity. Inhibitory effect of salicylates could be reversed by p50/p65 subunits of NF-kappaB or c-Jun overexpression. The inhibitory effect of aspirin on NF-kappaB activity was attributable to the inhibition of IkappaB kinase activity. Finally, tumors derived from C33A cells stably expressing LMP1 grown in nude mice showed enhanced MMP-9 levels compared with tumors derived from vector transfected C33A cells. This enhancement was inhibited by treatment of the mice with aspirin. These results suggest that aspirin may be able to suppress invasion and metastasis of EBV-associated tumors that express LMP1 by suppression of MMP 9. PMID- 10811141 TI - Reticulocyte cellular indices: a new approach in the diagnosis of anemias and monitoring of erythropoietic function. AB - Reticulocyte analysis has been extended from the simple enumeration of reticulocytes to precise measurements of mRNA content and of cellular indices such as volume, hemoglobin (Hb) concentration, and content. Assessment of reticulocyte maturity is based on the fluorescence intensity of reticulocytes, which depends on RNA content. The appearance of high fluorescence reticulocytes has been shown to be associated with engraftment in the setting of bone marrow or peripheral stem cells transplantation, although it is still not clear how this parameter can improve quality or cost of care compared with the traditional use of absolute neutrophil counts. Reticulocyte indices have been studied especially in the setting of iron deficiency and functional iron deficiency during recombinant human erythropoietin (r-HuEPO) therapy. Reticulocyte hemoglobin content (CHr) may allow prompt identification of an imbalance between r-HuEPO therapy and iron availability by detecting the presence in reticulocytes of iron restricted erythropoiesis. Diagnosis of simple iron deficiency can also be achieved in a more cost-effective fashion by using CHr in conjunction with the regular complete blood count (CBC), rather than relying on the traditional biochemical parameters of iron metabolism. Response to therapy of megaloblastic anemia can also be monitored with CHr. These new reticulocyte parameters provide a real-time assessment of the functional state of erythropoiesis. PMID- 10811142 TI - Measurement of cytokines in clinical samples using immunoassays: problems and pitfalls. AB - With the explosion in interest in cytokines and an increasing understanding of their possible involvement in the pathogenesis of many diseases, the number of studies measuring cytokines in body fluids has increased markedly over the last 5 to 10 years. However, it is readily apparent from the literature that the level of awareness of the many factors that can potentially influence the results obtained is very low. Such factors are likely to contribute considerably to the disparities seen among similar types of study. These include assay properties such as standardization and specificity, type of sample and sample handling, and many biological influences such as age, sex, ethnic background, and diurnal rhythmicity. Clearly, it is not possible to control for all these factors, but a minimum evaluation of assays should be carried out as described, including recovery and parallelity studies using relevant samples and the establishment of reproducibility and normal healthy population levels. An awareness of these factors, which is the purpose of this review, may hopefully lead to an improvement in the quality of studies incorporating the measurement of cytokines in clinical samples and should aid in the interpretation of the data. PMID- 10811143 TI - Studies on the gonadotropin-releasing activity of thymulin: changes with age. AB - We assessed the ability of thymulin, a zinc-dependent nonapeptide produced by the thymic epithelial cells, to influence the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from dispersed anterior pituitary (AP) cells from young, adult, and senescent female rats. Perifusion of young and senescent AP cells with thymulin doses of 10(-6) to 10(-5) M gave a significant stimulatory response for LH but not FSH. Gonadotropin release was always lower in the senescent cells. AP cells from both age groups incubated with 10(-8) to 10(-3) M thymulin showed a time- and dose-dependent response for both gonadotropins, with a maximal stimulation at 10(-7) M. Preincubation of thymulin with an antithymulin serum completely quenched the secretagogue activity of the hormone. Coincubation of thymulin with the secretagogue gonadotropin-releasing hormone (GnRH) revealed a synergistic effect on LH release and an additive effect on the release of FSH. The calcium chelator EGTA blocked the gonadotropin-releasing activity of thymulin in AP cells. The cAMP enhancers, caffeine, NaF, and forskolin significantly increased the thymulin-stimulated release of gonadotropins. The inositol phosphate enhancer LiCl potentiated the action of thymulin on gonadotropins. It is concluded that the gonadotropin-releasing activity documented here for thymulin is an age- and receptor-dependent effect mediated in part by calcium, cAMP, and inositol phosphates. PMID- 10811144 TI - Cardiovascular stress associated with concentric and eccentric isokinetic exercise in young and older adults. AB - Heart rate (HR), mean arterial blood pressure (MAP), and rate-pressure product (RPP) responses to submaximal isokinetic concentric (CON) and eccentric (ECC) knee extension exercise were compared at the same absolute torque output in 20 young (mean+/-SD=23.2+/-1.7 years) and 20 older (mean+/-SD=75.2+/-4.6 years) adults. After determination of peak CON and ECC torques, subjects performed separate, randomly ordered, 2-minute bouts of isokinetic CON and ECC exercise (90 degrees/s, exercise intensity: 50% of CON peak torque). CON exercise elicited greater changes in HR, MAP, and RPP than ECC exercise (p<.001) for both age groups. There were no age-related differences in HR, MAP, or RPP responses for either CON or ECC exercise. At the same absolute torque output, isokinetic CON knee extension exercise elicited significantly greater increases in cardiovascular stress than ECC exercise in both young and older adults. This result has implications for determining appropriate fitness and rehabilitation programs. PMID- 10811145 TI - Estriol (E3) replacement improves endothelial function and bone mineral density in very elderly women. AB - We investigated the effects of estriol (E3) on endothelial function and bone mineral density (BMD) in very elderly women. Twenty-four very elderly women (80 +/- 3.5 years old) were administered CaCl2 with or without estriol treatment (2 mg/day) for 30 weeks (hormone replacement treatment [HRT] group vs control group). Endothelium-dependent flow-mediated dilatation (FMD), endothelium independent dilatation by nitroglycerin of the brachial artery, and BMD were assessed. Levels of plasma lipids and apoproteins were not changed; however, both plasma E3 and E2 were substantially increased (E2, 4.6 to 31.3 +/- 8.1; E3, <5 to 45.3 +/- 7.9 pg/mL) by HRT. The FMD value was also increased by HRT, as were the plasma nitrite/nitrate and cGMP values. The response to nitroglycerin was not changed. The BMD was increased by HRT, but decreased in the control group. There were significant differences between the HRT group and control group after 30 weeks' treatment in the levels of osteocalcin, P1CP, and urinary deoxypiridinoridine. E3 significantly improved BMD by inhibiting bone resorption. Endothelial function was improved in line with the antiatherosclerotic effects. E3 might be effective for use in HRT in elderly patients. PMID- 10811146 TI - CBFA1 and topoisomerase I mRNA levels decline during cellular aging of human trabecular osteoblasts. AB - In order to understand the reasons for age-related impairment of the function of bone forming osteoblasts, we have examined the steady-state mRNA levels of the transcription factor CBFA1 and topoisomerase I during cellular aging of normal human trabecular osteoblasts, by the use of semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). There is a progressive and significant reduction of the CBFA1 steady-state mRNA level down to 50% during cellular aging of human osteoblasts. In comparison to the normal cells, human osteosarcoma cell lines SaOS-2 and KHOS/NP, and the SV40-transformed human lung fibroblast cell line MRC5V2 have 20 to 40% higher levels of CBFA1 mRNA. Similar levels of CBFA1 mRNA are detectable in normal human skin fibroblasts, and these cells also exhibit an age-related decline to the same extent. In addition, the expression of topoisomerase I is reduced by 40% in senescent osteoblasts, and the mRNA levels are significantly higher (40-70%) in transformed osteoblasts and fibroblasts. These changes in gene expression may be among the causes of impaired osteoblast functions, resulting in reduced bone formation during aging. PMID- 10811147 TI - Postreproductive life predicted by primate patterns. AB - Regression analyses of primate life spans on recently revised female body and brain masses of Old World primates predict a human life span of between 72 years and 91 years-estimates that exceed the age of human menopause (and prior estimates) by well over 20 years. The life spans predicted from body and brain sizes in the early Homo suggest that postreproductive life spans predate Homo sapiens Among anthropoid primates, residual longevity after body and brain effects are controlled is greatest for Homo and for the New World monkeys of the genus Cebus. Body and brain masses predict a 25-year life span for Cebus, although recorded life spans exceed 50 years. Cebus are geographically widespread, have a female-bonded social organization convergent with Old World monkeys, and are primarily frugivorous, though the diet is heavily supplemented with vertebrate prey. Regressions of phylogenetically independent contrasts indicate that body mass and brain mass relationships to longevity remain significant when phylogeny is controlled and that brain mass is a more robust predictor than body mass. These data are new in terms of the completeness of species representation, more reliable body masses, presentation of various comparison group regressions, and control for phylogenetic independence. PMID- 10811148 TI - Association of muscle power with functional status in community-dwelling elderly women. AB - BACKGROUND: Identification of the physiologic factors most relevant to functional independence in the elderly population is critical for the design of effective interventions. It has been suggested that muscle power may be more directly related to impaired physical performance than muscle strength in elderly persons. We tested the hypothesis that peak muscle power is closely associated with self reported functional status in sedentary elderly community-dwelling women. METHODS: We used baseline data that were collected as part of a 1-year randomized controlled clinical trial of a combined program of strength, power, and endurance training in 80 elderly women (mean age 74.8 +/- 5.0 years) with 3.2 +/- 1.9 chronic diseases, selected for baseline functional impairment and/or falls. RESULTS: Functional status at baseline was related in univariate analyses to physiologic capacity, habitual physical activity level, neuropsychological status, and medical diagnoses. Leg power had the strongest univariate correlation to self-reported functional status (r = -.47, p < .0001) of any of the physiologic factors we tested. In a forward stepwise regression model, leg press power and habitual physical activity level were the only two factors that contributed independently to functional status (r = .64, p < .0001), accounting for 40% of the variance in functional status. CONCLUSIONS: Leg power is a strong predictor of self-reported functional status in elderly women. PMID- 10811149 TI - Longitudinal course of behavioral problems during Alzheimer's disease: linear versus curvilinear patterns of decline. AB - BACKGROUND: Patients with Alzheimer's Disease (AD) are commonly assumed to experience a linear decline in behavioral functioning that parallels progressive cognitive decline. However, some researchers have suggested that specific behavioral problems either decline at different rates or improve in late dementia. METHODS: The present analyses examined 150 AD patients at an initial assessment, 61 of whom were also evaluated annually on two additional occasions. Measures of cognitive impairment and behavioral problems were obtained. RESULTS: Cross-sectional results indicated curvilinear associations between dementia severity and certain behavioral problems (forgetful behaviors, and emotional and impulsive behaviors). Longitudinal analyses further indicated trends for curvilinear rates of behavioral disturbance across time, with some problem areas showing improvement as AD progresses through the most severe stages. CONCLUSIONS: Even though Alzheimer's disease is a progressive dementia characterized by increasing cognitive deterioration, it appears to be inaccurate to expect behavioral functioning to show the same linear decline across time. PMID- 10811150 TI - Prevalence and psychosocial impact of urinary incontinence in older people of a Spanish rural population. AB - BACKGROUND: The prevalence and the psychosocial impact of urinary incontinence in older people of a rural population were investigated. METHODS: A cross-sectional study was made by means of a home health interview in a representative sample of people aged 65 years and older in the Basic Health Zone of Cabra (Cordoba). A questionnaire was used with two questions to detect urinary incontinence and further questions regarding its duration, frequency, volume, severity and psychosocial impact. Data were analyzed with basic statistics. A logistic regression model was created to identify which factors affect people's lifestyles. RESULTS: Among the 827 respondents the prevalence of incontinence reached 36% [95% confidence interval (CI): 34%-38%] and was higher (p < .001) in women (42%; 95% CI: 39%-45%) than in men (29%; 95% CI: 25%-38%). The most frequent duration of incontinence was from 1 to 5 years (16.8%). Fifteen percent had daily episodes and 10.6% estimated the volume of urine loss as high. Incontinence was severe in 9% of the affected subjects. Half the incontinent subjects expressed social limitations and negative feelings related to incontinence, and 21% admitted that incontinence negatively affected their lives. In the logistic regression model, both volume of urine loss [odds ratio (OR) = 5.61; 95% CI: 2.76-11.42] and being married (OR = 2.34; 95% CI: 1.03-5.33) were the factors statistically associated with a negative influence of incontinence on lifestyle. CONCLUSIONS: Urinary incontinence is a very frequent symptom in our population. Half the incontinent subjects suffer from some degree of psychosocial limitation. The volume of urine loss and being married are the most important factors with psychosocial impact. PMID- 10811151 TI - State variation in nursing home mortality outcomes according to do-not resuscitate status. AB - BACKGROUND: This study compares mortality outcomes of Medicaid-reimbursed nursing home residents with and without do-not-resuscitate (DNR) orders in two diverse states. METHODS: We used 1994 Minimum Data Set Plus (MDS+) information on 3215 nursing home residents from two states. We used Kaplan-Meier analyses to examine unadjusted mortality among those with and without DNR orders across states. We used a proportional hazard regression with main and interaction variables to model the likelihood of survival in the nursing home. RESULTS: Approximately 27% of nursing home residents with DNR orders in State A die within the year, and approximately 40% of nursing home residents with DNR orders in State B die within the year. Regression results indicate that neither having a DNR order nor state of residence were independently associated with mortality. However, residing in State B and having a DNR order was associated with an increased risk of mortality compared with all others in the sample (risk ratio = 1.73; 95% confidence interval = 1.09, 2.75). CONCLUSION: This study demonstrates that DNR orders are associated with varying mortality across states. Future research is needed to identify the reasons why state level differences exist. PMID- 10811152 TI - Lower extremity function and subsequent disability: consistency across studies, predictive models, and value of gait speed alone compared with the short physical performance battery. AB - BACKGROUND: Although it has been demonstrated that physical performance measures predict incident disability in previously nondisabled older persons, the available data have not been fully developed to create usable methods for determining risk profiles in community-dwelling populations. Using several populations and different follow-up periods, this study replicates previous findings by using the Established Populations for the Epidemiologic Study of the Elderly (EPESE) performance battery and provides equations for the prediction of disability risk according to age, sex, and level of performance. METHODS: Tests of balance, time to walk 8 ft, and time to rise from a chair 5 times were administered to 4,588 initially nondisabled persons in the four sites of the EPESE and to 1,946 initially nondisabled persons in the Hispanic EPESE. Follow-up assessment for activity of daily living (ADL) and mobility-related disability occurred from 1 to 6 years later. RESULTS: In the EPESE, compared with those with the best performance (EPESE summary performance score of 10-12), the relative risks of mobility-related disability for those with scores of 4-6 ranged from 2.9 to 4.9 and the relative risk of disability for those with scores of 7-9 ranged from 1.5 to 2.1, with similar consistent results for ADL disability. The observed rates of incident disability according to performance level in the Hispanic EPESE agreed closely with rates predicted from models developed from the EPESE sites. Receiver operating characteristic curves showed that gait speed alone performed almost as well as the full battery in predicting incident disability. CONCLUSIONS: Performance tests of lower extremity function accurately predict disability across diverse populations. Equations derived from models using both the summary score and the gait speed alone allow for the estimation of risk of disability in community-dwelling populations and provide valuable information for estimating sample size for clinical trials of disability prevention. PMID- 10811154 TI - The effect of body position on arterial oxygen saturation in acute stroke. AB - BACKGROUND: Evidence suggests that respiratory function is impaired poststroke. Body position is known to influence respiratory function in normal subjects and those with respiratory pathologies. Its effect on respiratory function after stroke has received little attention. However, one study suggests that some positions used in clinical practice may adversely influence respiratory function. This study therefore aimed to identify resting positions that maintain arterial oxygen saturation (SaO2) at optimal levels, changes in SaO2 during time spent in the test position, and differences in SaO2 among the positions investigated. METHOD: A within-subject, two-center clinical study was made. Patients in the first 72 hours following mild to moderately severe stroke were allocated a randomized sequence of four positions. One hour was spent in each position. SaO2 was recorded each minute by pulse oximetry with a finger probe. Mean values for the hour were calculated. RESULTS: Mean arterial oxygen saturation values for all patients were >90% for the hour spent in each test position for all patients. There were no changes in arterial oxygen saturation across the hour spent in the test positions (repeated-measures analysis of variance). No differences in arterial oxygen saturation were identified among positions (analysis of covariance). DISCUSSION: The saturation levels recorded corresponded to those observed in studies of normal elderly persons. The positions tested may be recommended for use in clinical practice to maintain arterial oxygen saturation in patients in the first 72 hours following mild to moderately severe stroke. PMID- 10811153 TI - Older age and in-hospital development of hypokalemia from loop diuretics: results from a multicenter survey. GIFA Investigators. Multicenter Italian Pharmacoepidemiologic Study Group. AB - BACKGROUND: Hypokalemia is a common finding among older patients taking diuretic medications. However, it is not known whether older age per se carries an increased risk of hypokalemia, particularly during a patient's treatment with loop diuretics. METHODS: The association between age and incident hypokalemia was examined in 18,872 patients with normal baseline serum potassium enrolled during three yearly multicenter surveys; 4,035 patients started receiving loop diuretics during their hospital stay. Demographic variables, comorbid conditions, medications, and objective tests that were associated with incident hypokalemia in separate age- and sex-adjusted logistic regression models were examined as potential confounders. RESULTS: Among patients with normal baseline serum potassium, the factors of age, presence of coronary disease or diabetes, comorbidity, the use of ACE inhibitors, loop diuretics, digitalis, corticosteroids, or insulin, and baseline serum potassium were associated with incident hypokalemia in initial models. After these variables were adjusted for, age (for each decade, odds ratio = 1.30; 95% confidence interval = 1.17-1.46; p < .0001) was associated with incident hypokalemia. The use of parenteral (2.30; 1.53-3.46; p < .0001) but not oral (1.16; 0.79-1.69; p = .44) loop diuretics was associated with hypokalemia. Eventually, age was associated with hypokalemia when the summary regression model was analyzed in patients taking loop diuretics (1.33; 1.03-1.71; p = .027), as well as in those taking intravenous loop diuretics only (1.84; 1.25-2.70; p = .002). CONCLUSIONS: Older age is independently associated with the in-hospital development of hypokalemia, particularly among patients taking loop diuretics. Monitoring of serum potassium levels is therefore advisable when older patients are treated with these agents. PMID- 10811155 TI - Effect of endurance exercise training on left ventricular size and remodeling in older adults with hypertension. AB - BACKGROUND: It is not known whether exercise training can induce a reduction of blood pressure (BP) and a regression of left ventricular hypertrophy (LVH) in older hypertensive subjects. This study was designed to determine whether endurance exercise training, by lowering BP, can induce regression of LVH and left ventricular (LV) concentric remodeling in older hypertensive adults. METHODS: We studied 11 older adults with mild to moderate hypertension (BP 152.0 +/- 2.5/91.3 +/- 1.5 mm Hg, mean +/- SE), 65.5 +/- 1.2 years old, who exercised for 6.8 +/- 3.8 months. Seven sedentary hypertensive (BP 153 +/- 3/89 +/- 2 mm Hg) subjects, 68.5 +/- 1 years old, served as controls. LV size and geometry and function were assessed with the use of two-dimensional echocardiography. RESULTS: Exercise training increased aerobic power by 16% (p < .001), and it decreased systolic (p < .05) and diastolic (p < .05) BP, LV wall thickness (from 12.8 +/- 0.4 mm to 11.3 +/- 0.3 mm; p < .05), and the wall thickness-to-radius (h/r) ratio (from 0.48 +/- 0.02 to 0.41 +/- 0.01; p < .05). There were no significant changes in the controls. The changes in LV mass index (deltaLVMI) were different between the two groups. LV mass index decreased in the exercise group (deltaLVMI - 14.3 +/- 3.3 g) but not in the controls (deltaLVMI 1.4 +/- 4.1 g; p = .009). A multiple stepwise regression analysis showed that among clinical and physiological variables including changes in resting systolic BP, aerobic power, body mass index, and systolic BP during submaximal and maximal exercise, only the reduction in resting systolic BP correlated significantly with a regression of concentric remodeling (delta h/r ratio r = .80; p = .003). The other variables did not add to the ability of the model to predict changes in the h/r ratio. CONCLUSIONS: The data suggest that exercise training can reduce BP and induce partial regression of LVH and LV concentric remodeling in older adults with mild or moderate hypertension. PMID- 10811156 TI - Minimal information to determine affine shape equivalence. AB - Participants judged the affine equivalence of 2 simultaneously presented 4-point patterns. Performance level (d') varied between 1.5 and 2.7, depending on the information available for solving the correspondence problem (insufficient in Experiment 1a, superfluous in Experiment 1b, and minimal in Experiments 1c, 2a, 2b) and on the exposure time (unlimited in Experiments 1 and 2a and 500 ms in Experiment 2b), but it did not vary much with the complexity of the affine transformation (rotation and slant in Experiment 1 and same plus tilt in Experiment 2). Performance in Experiment 3 was lower with 3-point patterns than with 4-point patterns, whereas blocking the trials according to the affine transformation parameters had little effect. Determining affine shape equivalence with minimal-information displays is based on a fast assessment of qualitatively or quasi-invariant properties such as convexity/ concavity, parallelism, and collinearity. PMID- 10811157 TI - Word-based grouping affects the prime-task effect on semantic priming. AB - Semantic priming between words is reduced or eliminated if a low-level task such as letter search is performed on the prime word (the prime task effect), a finding used to question the automaticity of semantic processing of words. This idea is critically examined in 3 experiments with a new design that allows the search target to occur both inside and outside the prime word. The new design produces the prime task effect (Experiment 1) but shows semantic negative priming when the target letter occurs outside the prime word (Experiments 2 and 3). It is proposed that semantic activation and priming are dissociable and that inhibition and word-based grouping are responsible for reduction of semantic priming in the prime task effect. PMID- 10811158 TI - Does unattended information facilitate change detection? AB - Changes between alternating visual displays are difficult to detect when the successive presentations of the displays are separated by a brief temporal interval. To assess whether unattended changes attract attention, observers searched for the location of a change involving either a large or a small number of features, in pairs of displays consisting of 4, 7, 10, 13, or 16 letters (Experiment 1) or digits (Experiments 2 and 3). Each display in a pair of displays was presented for 200 ms, and either a blank screen (Experiments 1 and 2) or a screen of equal luminance to the letters and digits (Experiment 3) was presented for 80 ms between the alternating displays. In all experiments, the search function for locating the larger change was shallower than the search function for locating the smaller change. These results indicate that unattended changes play a functional role in guiding focal attention. PMID- 10811159 TI - Attention to object files defined by transparent motion. AB - Two interspersed and differently colored sets of dots were rotated in opposite directions and were perceived as superimposed transparent surfaces. Probes consisting of brief changes in dot motion direction were reported. Two probes affecting the same surface were discriminated accurately. The 2nd probe was discriminated poorly if it affected a surface different from the 1st and if the time between probes was less than 600 ms. This reflects a difficulty in switching attention rapidly between surfaces. Spatial proximity increased the interference. Controls were incompatible with traditional spatial mechanisms (2- or 3 dimensional) or with simple sensory filters. Instead, probes were apparently selected by object files. The interference is not simply due to an inability to process 2 objects at once but requires close spatial proximity of incompatible motion signals. PMID- 10811160 TI - Levels of selective attention revealed through analyses of response time distributions. AB - The present research examines the nature of the interference effects in a number of selective attention tasks. All of these tasks result in interference in performance by presenting information that is irrelevant to task performance but competes for selection. The interference from this competing information slows the response time (RT) of participants relative to a condition where the competition is minimized. The authors use a convolution of an exponential and a Gaussian (ex-Gaussian) distribution to examine the influence of interference on the characteristics of RT distributions. Consistent with previous research, the authors show that interference in the Stroop task is reflected by both the Gaussian and exponential portions of the ex-Gaussian. In contrast, in 4 experiments they show that several other interference tasks evidence interference that is reflected only in the Gaussian portion of the ex-Gaussian distribution. The authors suggest that these differences reflect the operation of different selection mechanisms, and they examine how sequential sampling models accommodate these effects. PMID- 10811161 TI - Configural information in facial expression perception. AB - Composite facial expressions were prepared by aligning the top half of one expression (e.g., anger) with the bottom half of another (e.g., happiness). Experiment 1 shows that participants are slower to identify the expression in either half of these composite images relative to a "noncomposite" control condition in which the 2 halves are misaligned. This parallels the composite effect for facial identity (A. W. Young, D. Hellawell, & D. C. Hay, 1987), and like its identity counterpart, the effect is disrupted by inverting the stimuli (Experiment 2). Experiment 3 shows that no composite effect is found when the top and bottom sections contain different models' faces posing the same expression; this serves to exclude many nonconfigural interpretations of the composite effect (e.g., that composites are more "attention-grabbing" than noncomposites). Finally, Experiment 4 demonstrates that the composite effects for identity and expression operate independently of one another. PMID- 10811162 TI - Geometrical illusions can affect time-to-contact estimation and mimed prehension. AB - Results indicate that under some conditions the Sander parallelogram illusion can affect time-to-contact (TTC) estimation in a prediction-motion (PM) task and in an interceptive action (IA). The illusion also affected mimed manual prehension. The implication is that the timing of responses in the PM and IA tasks may involve an estimate of TTC that is based on the perceived dimensions of the environment. Further research is warranted in the development of models of perceived collision and of visually guided actions. PMID- 10811163 TI - The combined effects of plane disorientation and foreshortening on picture naming: one manipulation or two? AB - Objects disoriented in plane away from the upright and objects rotated in depth producing foreshortening are harder to identify than canonical views. In Experiments 1 and 2, participants named pictures of familiar objects. There was no interaction between plane and depth rotation effects on initial presentation or after practice. Experiment 3 was a dual-task psychological refractory period study. Participants classified a high-low tone with a speeded keypress and then named a canonical, plane-rotated, or foreshortened view of an object. Naming was slower when the picture was presented 50 ms after the tone compared with 800 ms after the tone. Plane rotation effects were reduced (but not eliminated) at the short tone-picture stimulus onset asynchrony, but foreshortening effects were not reduced. The results implicate an early, prebottleneck locus for some processes compensating for plane rotation and a subsequent bottleneck or postbottleneck locus for compensation for foreshortening. PMID- 10811164 TI - Eye height scaling of absolute size in immersive and nonimmersive displays. AB - Eye-height (EH) scaling of absolute height was investigated in three experiments. In Experiment 1, standing observers viewed cubes in an immersive virtual environment. Observers' center of projection was placed at actual EH and at 0.7 times actual EH. Observers' size judgments revealed that the EH manipulation was 76.8% effective. In Experiment 2, seated observers viewed the same cubes on an interactive desktop display; however, no effect of EH was found in response to the simulated EH manipulation. Experiment 3 tested standing observers in the immersive environment with the field of view reduced to match that of the desktop. Comparable to Experiment 1, the effect of EH was 77%. These results suggest that EH scaling is not generally used when people view an interactive desktop display because the altitude of the center of projection is indeterminate. EH scaling is spontaneously evoked, however, in immersive environments. PMID- 10811165 TI - Contingent capture for onsets and offsets: attentional set for perceptual transients. AB - Four experiments were conducted to examine whether attentional set affects the ability of visual transients (onsets and offsets) to capture attention. In the experiments, visual search for an identity-defined target was conducted. In the first 3 experiments, the target display either onset entirely or was revealed by offsetting camouflaging line segments to reveal letters. Prior to the target display, there was a noninformative cue, either an onset or an offset, at one of the potential target locations. Cues that shared the same transient feature as the target display captured attention. The lack of predictable target transients led to attentional capture by all forms of transients. The final experiments with luminance changes without offsets or onsets showed attentional capture when the luminance changes were large. The results suggest that attentional set can be broadly or narrowly tuned to detect changes in luminance. PMID- 10811166 TI - The role of phonological codes in integrating information across saccadic eye movements in Chinese character identification. AB - Prior research has generally assumed either that phonological codes do not contribute to Chinese character identification or that they do so only through a look-up process at the character level. In 3 experiments, a homophone seen parafoveally aided the identification of a target character that was fixated following an eye movement to the preview location. Moreover, high-frequency phonetically regular characters were named faster than high-frequency, phonetically irregular characters. Thus, both lexical and sublexical phonological codes of Chinese characters are involved early in the process of character identification. Orthographic information from the preview was also used in character identification, as orthographically similar previews facilitated target identification as well. The evidence for the extraction of semantic information from parafoveal previews was mixed, as synonym previews facilitated in Experiment 2 but not in Experiment 1. PMID- 10811167 TI - Motor activation from visible speech: evidence from stimulus response compatibility. AB - In speech perception, phonetic information can be acquired optically as well as acoustically. The motor theory of speech perception holds that motor control structures are involved in the processing of visible speech, whereas perceptual accounts do not make this assumption. Motor involvement in speech perception was examined by showing participants response-irrelevant movies of a mouth articulating /ba/ or /da/ and asking them to verbally respond with either the same or a different syllable. The letters "Ba" and "Da" appeared on the speaker's mouth to indicate which response was to be performed. A reliable interference effect was observed. In subsequent experiments, perceptual interference was ruled out by using response-unrelated imperative stimuli and by preexposing the relevant stimulus information. Further, it was demonstrated that simple directional features (opening and closing) do not account for the effect. Rather, the present study provides evidence for the view that visible speech is processed up to a late, response-related processing stage, as predicted by the motor theory of speech perception. PMID- 10811168 TI - The role of long-term-memory and short-term-memory links in the Simon effect. AB - In Experiment 1, children performed a Simon task after a spatially compatible or incompatible task. Results showed a Simon effect after the spatially compatible task and a reversed Simon effect after the spatially incompatible task. In Experiments 2-5, an identical procedure was adopted with adult participants, who performed the Simon task immediately after, a day after, or a week after the spatial compatibility task. Experiment 6 established a baseline for the Simon effect. Results showed a Simon effect after the spatially compatible task and no Simon effect or a reversed Simon effect after the spatially incompatible task. A modified version of the computational model of M. Zorzi and C. Umilta (1995) was used to compare possible accounts of the findings. The best account exploits 2 types of short-term-memory links between stimulus and response and their interaction with long-term-memory links. PMID- 10811169 TI - Recruitment of degrees of freedom stabilizes coordination. AB - By showing that transitions may be obviated by recruiting degrees of freedom in the coupled pendulum paradigm, the authors reveal a novel mechanism for coordinative flexibility. In Experiment 1, participants swung pairs of unconstrained pendulums in 2 planes of motion (sagittal and frontal) at 8 movement frequencies starting from either an in-phase or antiphase mode. Few transitions were observed. Measures of spatial trajectory showed recruitment effects tied to the stability of the initial coordinative pattern. When the motion of the pendulums was physically restricted to a single plane in Experiment 2, transitions were more common, indicating that recruitment delays--or even eliminates--transitions. Such recruitment complements transitions as a source of coordinative flexibility and is incorporated in a simple extension of the Haken Kelso-Bunz (1985) model. PMID- 10811170 TI - Postattentive vision. AB - Much research has examined preattentive vision: visual representation prior to the arrival of attention. Most vision research concerns attended visual stimuli; very little research has considered postattentive vision. What is the visual representation of a previously attended object once attention is deployed elsewhere? The authors argue that perceptual effects of attention vanish once attention is redeployed. Experiments 1-6 were visual search studies. In standard search, participants looked for a target item among distractor items. On each trial, a new search display was presented. These tasks were compared to repeated search tasks in which the search display was not changed. On successive trials, participants searched the same display for new targets. Results showed that if search was inefficient when participants searched a display the first time, it was inefficient when the same, unchanging display was searched the second, fifth, or 350th time. Experiments 7 and 8 made a similar point with a curve tracing paradigm. The results have implications for an understanding of scene perception, change detection, and the relationship of vision to memory. PMID- 10811171 TI - Auditory spatial attention using interaural time differences. AB - Previous probe-signal studies of auditory spatial attention have shown faster responses to sounds at an expected versus an unexpected location, making no distinction between the use of interaural time difference (ITD) cues and interaural-level difference cues. In 5 experiments, performance on a same different spatial discrimination task was used in place of the reaction time metric, and sounds, presented over headphones, were lateralized only by an ITD. In all experiments, performance was better for signals lateralized on the expected side of the head, supporting the conclusion that ITDs can be used as a basis for covert orienting. The performance advantage generalized to all sounds within the spatial focus and was not dissipated by a trial-by-trial rove in frequency or by a rove in spectral profile. Successful use by the listeners of a cross-modal, centrally positioned visual cue provided evidence for top-down attentional control. PMID- 10811173 TI - You must see the point: automatic processing of cues to the direction of social attention. AB - Four experiments explored the processing of pointing gestures comprising hand and combined head and gaze cues to direction. The cross-modal interference effect exerted by pointing hand gestures on the processing of spoken directional words, first noted by S. R. H. Langton, C. O'Malley, and V. Bruce (1996), was found to be moderated by the orientation of the gesturer's head-gaze (Experiment 1). Hand and head cues also produced bidirectional interference effects in a within modalities version of the task (Experiment 2). These findings suggest that both head-gaze and hand cues to direction are processed automatically and in parallel up to a stage in processing where a directional decision is computed. In support of this model, head-gaze cues produced no influence on nondirectional decisions to social emblematic gestures in Experiment 3 but exerted significant interference effects on directional responses to arrows in Experiment 4. It is suggested that the automatic analysis of head, gaze, and pointing gestures occurs because these directional signals are processed as cues to the direction of another individual's social attention. PMID- 10811172 TI - Understanding projectile acceleration. AB - Throwing and catching balls or other objects is a generally highly practiced skill; however, conceptual as well as perceptual understanding of the mechanics that underlie this skill is surprisingly poor. In 5 experiments, we investigated conceptual and perceptual understanding of simple ballistic motion. Paper-and pencil tests revealed that up to half of all participants mistakenly believed that a ball would continue to accelerate after it left the thrower's hand. Observers also showed a remarkable tolerance for anomalous trajectory shapes. Perceptual judgments based on graphics animations replicated these erroneous beliefs for shallow release angles. Observers' tolerance for anomalies tended to decrease with their distance from the actor. The findings are at odds with claims of the naive physics literature that liken intuitive understanding to Aristotelian or medieval physics theories. Instead, observers seem to project their intentions to the ball itself (externalization) or even feel that they have power over the ball when it is still close. PMID- 10811174 TI - Syllables in the processing of spoken Italian. AB - Five experiments explored the role of the syllable in the processing of spoken Italian. According to the syllabic hypothesis, the sublexical unit used by speakers of Romance languages to segment speech and access the lexicon is the syllable. However, languages with different degrees of acoustic-phonetic transparency give rise to syllabic effects that vary in robustness. It follows from this account that speakers of phonologically similar languages should behave in a similar way. By exploiting the similarities between Spanish and Italian, the authors tested this prediction in Experiments 1-4. Indeed, Italian listeners were found to produce syllabic effects similar to those observed in Spanish listeners. In Experiment 5, the predictions of the syllabic hypothesis with respect to lexical access were tested. The results corroborated these predictions. The findings are discussed in relation to current models of speech processing. PMID- 10811175 TI - Oculocentric coding of inhibited eye movements to recently attended locations. AB - Results are reported for experiments that examined eye movements directed toward recently cued objects. In 1 experiment participants were slower to initiate saccades toward the earlier location of an object that had been cued, even though the cued object had subsequently moved away from that location. Other experiments involved exploring the reference frame within which the inhibited eye movements are encoded. These experiments revealed that the eye movement that is inhibited is encoded in an oculocentric-rather than an environmental-reference frame. However, simple detection as indexed by manual keypress responses is encoded in an environmental reference frame. The results have implications for inhibition of return, for the link between eye movements and attention, and for the nature of the spatial reference frames in which both covert and overt movements of attention are encoded. PMID- 10811176 TI - Relative judgment seems to be the key: revisiting the Beck effect. AB - In multiple-stimulus presentation, orientation disparity has been known to be more discriminable than disparity in line arrangement (e.g., J. Beck, 1972). The source of the effect and its locus were studied in 7 experiments. In different experiments a discrimination between an upright T and either a tilted T or an L, or a discrimination between a tilted T and an L, was required, either in a single stimulus presentation or in the context of upright Ts. Number of stimuli, location uncertainty, and adjacency between stimuli were manipulated. The results indicated that the effect is insensitive to these factors, which is incommensurate with predictions from several accounts of the effect. All the effect requires is that disparate stimuli are simultaneously presented, suggesting that relative judgment is a necessary condition for its manifestation. The effect surfaces when the task calls for procedures based on perception of homogeneity or salience. PMID- 10811177 TI - Face and mouth inversion effects on visual and audiovisual speech perception. AB - Three experiments examined whether image manipulations known to disrupt face perception also disrupt visual speech perception. Research has shown that an upright face with an inverted mouth looks strikingly grotesque whereas an inverted face and an inverted face containing an upright mouth look relatively normal. The current study examined whether a similar sensitivity to upright facial context plays a role in visual speech perception. Visual and audiovisual syllable identification tasks were tested under 4 presentation conditions: upright face-upright mouth, inverted face-inverted mouth, inverted face-upright mouth, and upright face-inverted mouth. Results revealed that for some visual syllables only the upright face-inverted mouth image disrupted identification. These results suggest that upright facial context can play a role in visual speech perception. A follow-up experiment testing isolated mouths supported this conclusion. PMID- 10811178 TI - The role of morphological constituents in reading Finnish compound words. AB - The processing of transparent Finnish compound words was investigated in 2 experiments in which eye movements were recorded while sentences were read silently. The frequency of the second constituent had a large influence (95 ms) on gaze duration on the target words, but its influence was relatively late in processing: A clear effect only occurred on the probability of a third fixation. The frequency of the whole compound word had a similar influence on gaze duration (82 ms) and influenced eye movements at least as rapidly as did the frequency of the second constituent. These results, together with an earlier finding that the frequency of the first constituent affected the first fixation duration, indicate that the identification of these compound words involves parallel processing of both morphological constituents and whole-word representations. PMID- 10811179 TI - Evidence for split attentional foci. AB - A partial report procedure was used to test the ability of observers to split attention over noncontiguous locations. Observers reported the identity of 2 targets that appeared within a 5 x 5 stimulus array, and cues (validity = 80%) informed them of the 2 most likely target locations. On invalid trials, 1 of the targets appeared directly in between the cued locations. Experiments 1, 1a, and 2 showed a strong accuracy advantage at cued locations compared with intervening ones. This effect was larger when the cues were arranged horizontally rather than vertically. Experiment 3 suggests that this effect of cue orientation reflects an advantage for processing targets that appear in different hemifields. Experiments 4 and 4a suggest that the primary mechanism supporting the flexible deployment of spatial attention is the suppression of interference from stimuli at unattended locations. PMID- 10811181 TI - Stem-cell research and the US Congress. PMID- 10811180 TI - Serial processing in reading aloud: no challenge for a parallel model. AB - K. Rastle and M. Coltheart (1999) challenged parallel models of reading by showing that the cost of irregularity in low-frequency exception words was modulated by the position of the irregularity in the word. This position-of irregularity effect was taken as strong evidence of serial processing in reading. This article refutes Rastle and Coltheart's theoretical conclusions in 3 ways: First, a parallel model, the connectionist dual process model (M. Zorzi, G. Houghton, & B. Butterworth, 1998b), produces a position-of-irregularity effect. Second, the supposed serial effect can be reduced to a position-specific grapheme phoneme consistency effect. Third, the position-of-irregularity effect vanishes when the experimental data are reanalyzed using grapheme-phoneme consistency as the covariate. This demonstration has broader implications for studies aiming at adjudicating between models: Strong inferences should be avoided until the computational models are actually tested. PMID- 10811182 TI - Agricultural research whistling in the dark. PMID- 10811184 TI - Doubts and uncertainties slow NASA's schedule PMID- 10811183 TI - NIH opposes plans for patenting 'similar' gene sequences. PMID- 10811185 TI - New panel set to beat biotech's bad image... PMID- 10811187 TI - CJD survey offers Britain a glimmer of hope. PMID- 10811186 TI - Battle lines shift in stem-cell funding fight...as non-profit companies enter the fray. PMID- 10811189 TI - NSF celebrates half a century with big plans PMID- 10811188 TI - Space-station airlock to serve as temporary lab PMID- 10811190 TI - US treats AIDS as a threat to security. PMID- 10811192 TI - Sagan breached security by revealing US work on a lunar bomb project PMID- 10811193 TI - Austria takes lab animal welfare seriously. PMID- 10811191 TI - The missionary from Munich. PMID- 10811194 TI - How metrics could have saved UK car industry PMID- 10811195 TI - Instrument's ability to do the job is NASA's priority PMID- 10811197 TI - Persian role in flowering of Islamic science PMID- 10811196 TI - Putting marine mammals back in the mainstream. PMID- 10811198 TI - Oeconomia Naturae L PMID- 10811199 TI - Et in articulo mortis PMID- 10811200 TI - Evolution all at sea. PMID- 10811201 TI - Palaeoanthropology. Coasting out of Africa. PMID- 10811202 TI - Fluid dynamics. Turbulence without inertia PMID- 10811204 TI - Earth science. Strength of the San Andreas PMID- 10811203 TI - Apoptosis. Gone but not forgotten. PMID- 10811206 TI - Beetle larvae cooperate to mimic bees. PMID- 10811205 TI - A single birth of all plastids? PMID- 10811207 TI - Coral bleach-out in Belize. PMID- 10811208 TI - Transgenerational mutation by radiation. PMID- 10811209 TI - Spontaneous formation of inorganic helices PMID- 10811210 TI - A surprising simplicity to protein folding. AB - The polypeptide chains that make up proteins have thousands of atoms and hence millions of possible inter-atomic interactions. It might be supposed that the resulting complexity would make prediction of protein structure and protein folding mechanisms nearly impossible. But the fundamental physics underlying folding may be much simpler than this complexity would lead us to expect folding rates and mechanisms appear to be largely determined by the topology of the native (folded) state, and new methods have shown great promise in predicting protein-folding mechanisms and the three-dimensional structures of proteins. PMID- 10811211 TI - Molecular evidence for genetic mixing of Arctic and Antarctic subpolar populations of planktonic foraminifers. AB - Bipolarity, the presence of a species in the high latitudes separated by a gap in distribution across the tropics, is a well-known pattern of global species distribution. But the question of whether bipolar species have evolved independently at the poles since the establishment of the cold-water provinces 16 8 million years ago, or if genes have been transferred across the tropics since that time, has not been addressed. Here we examine genetic variation in the small subunit ribosomal RNA gene of three bipolar planktonic foraminiferal morphospecies. We identify at least one identical genotype in all three morphospecies in both the Arctic and Antarctic subpolar provinces, indicating that trans-tropical gene flow must have occurred. Our genetic analysis also reveals that foraminiferal morphospecies can consist of a complex of genetic types. Such occurrences of genetically distinct populations within one morphospecies may affect the use of planktonic foraminifers as a palaeoceanographic proxy for climate change and necessitate a reassessment of the species concept for the group. PMID- 10811212 TI - Io as a source of the jovian dust streams AB - Streams of dust emerging from the direction of Jupiter were discovered in 1992 during the flyby of the Ulysses spacecraft, but their precise origin within the jovian system remained unclear. Further data collected by the Galileo spacecraft, which has been orbiting Jupiter since December 1995, identified the possible sources of dust as Jupiter's main ring, its gossamer ring, comet Shoemaker-Levy 9 (ref. 8) and Io. All but Jupiter's gossamer ring and Io have since been ruled out. Here we find that the dominant source of the jovian dust streams is Io, on the basis of periodicities in the dust impact signal. Io's volcanoes, rather than impact ejecta, are the dust sources. PMID- 10811213 TI - Detection of doubled shot noise in short normal-metal/superconductor junctions AB - Shot noise refers to the fluctuations in electrical current through a device arising from the discrete nature of the charge-carrying particles. Recent experiments have exploited the fact that the shot noise is proportional to the charge of the carriers to establish fractional quantization of quasiparticles in the fractional quantum Hall effect. By a similar argument, it is expected that when a superconducting reservoir emits Cooper pairs, (which have a charge twice that of an electron) into a short normal-metal wire, the shot noise should be double that obtained for a normal-metal reservoir. Although the charge of Cooper pairs has been well established by flux quantization and tunnel experiments, doubling of their shot noise has not yet been observed. Here we report a shot noise experiment using a short diffusive normal-metal superconductor contact, in which we confirm the predicted noise behaviour for double charges. The measurements, taken over a large range of bias current, establish that phase coherence is not required to observe the effect. PMID- 10811214 TI - Elastic turbulence in a polymer solution flow AB - Turbulence is a ubiquitous phenomenon that is not fully understood. It is known that the flow of a simple, newtonian fluid is likely to be turbulent when the Reynolds number is large (typically when the velocity is high, the viscosity is low and the size of the tank is large). In contrast, viscoelastic fluids such as solutions of flexible long-chain polymers have nonlinear mechanical properties and therefore may be expected to behave differently. Here we observe experimentally that the flow of a sufficiently elastic polymer solution can become irregular even at low velocity, high viscosity and in a small tank. The fluid motion is excited in a broad range of spatial and temporal scales, and we observe an increase in the flow resistance by a factor of about twenty. Although the Reynolds number may be arbitrarily low, the observed flow has all the main features of developed turbulence. A comparable state of turbulent flow for a newtonian fluid in a pipe would have a Reynolds number as high as 10(5) (refs 1, 2). The low Reynolds number or 'elastic' turbulence that we observe is accompanied by significant stretching of the polymer molecules, resulting in an increase in the elastic stresses of up to two orders of magnitude. PMID- 10811215 TI - Rapid prototyping of patterned functional nanostructures AB - Living systems exhibit form and function on multiple length scales and at multiple locations. In order to mimic such natural structures, it is necessary to develop efficient strategies for assembling hierarchical materials. Conventional photolithography, although ubiquitous in the fabrication of microelectronics and microelectromechanical systems, is impractical for defining feature sizes below 0.1 micrometres and poorly suited to pattern chemical functionality. Recently, so called 'soft' lithographic approaches have been combined with surfactant and particulate templating procedures to create materials with multiple levels of structural order. But the materials thus formed have been limited primarily to oxides with no specific functionality, and the associated processing times have ranged from hours to days. Here, using a self-assembling 'ink' we combine silica surfactant self-assembly with three rapid printing procedures--pen lithography, ink-jet printing, and dip-coating of patterned self-assembled monolayers--to form functional, hierarchically organized structures in seconds. The rapid-prototyping procedures we describe are simple, employ readily available equipment, and provide a link between computer-aided design and self-assembled nanostructures. We expect that the ability to form arbitrary functional designs on arbitrary surfaces will be of practical importance for directly writing sensor arrays and fluidic or photonic systems. PMID- 10811216 TI - Large differences in tropical aerosol forcing at the top of the atmosphere and Earth's surface AB - The effect of radiative forcing by anthropogenic aerosols is one of the largest sources of uncertainty in climate predictions. Direct observations of the forcing are therefore needed, particularly for the poorly understood tropical aerosols. Here we present an observational method for quantifying aerosol forcing to within +/-5 per cent. We use calibrated satellite radiation measurements and five independent surface radiometers to quantify the aerosol forcing simultaneously at the Earth's surface and the top of the atmosphere over the tropical northern Indian Ocean. In winter, this region is covered by anthropogenic aerosols of sulphate, nitrate, organics, soot and fly ash from the south Asian continent. Accordingly, mean clear-sky solar radiative heating for the winters of 1998 and 1999 decreased at the ocean surface by 12 to 30 Wm(-2), but only by 4 to 10 Wm( 2) at the top of the atmosphere. This threefold difference (due largely to solar absorption by soot) and the large magnitude of the observed surface forcing both imply that tropical aerosols might slow down the hydrological cycle. PMID- 10811217 TI - Sensitivity of the geomagnetic axial dipole to thermal core-mantle interactions AB - Since the work of William Gilbert in 1600 (ref. 1), it has been widely believed that the Earth's magnetic field, when suitably time-averaged, is that of a magnetic dipole positioned at the Earth's centre and aligned with the rotational axis. This 'geocentric axial dipole' (GAD) hypothesis has been the central model for the study of the Earth's magnetic field--it underpins almost all interpretations of palaeomagnetic data, whether for studies of palaeomagnetic secular variation, for plate tectonic reconstructions, or for studies of palaeoclimate. Although the GAD hypothesis appears to provide a good description of the Earth's magnetic field over at least the past 100 Myr (ref. 2), it is difficult to test the hypothesis for earlier periods, and there is some evidence that a more complicated model is required for the period before 250 Myr ago. Kent and Smethurst suggested that this additional complexity might be because the inner core would have been smaller at that time. Here I use a numerical geodynamo model and find that reducing the size of the inner core does not significantly change the character of the magnetic field. I also consider an alternative process that could lead to the breakdown of the GAD hypothesis on this timescale, the evolution of heat-flux variations at the core-mantle boundary, induced by mantle convection. I find that a simple pattern of heat-flux variations at the core-mantle boundary, which is plausible for times before the Mesozoic era, results in a strong octupolar contribution to the field, consistent with previous findings. PMID- 10811218 TI - Early human occupation of the Red Sea coast of Eritrea during the last interglacial. AB - The geographical origin of modern humans is the subject of ongoing scientific debate. The 'multiregional evolution' hypothesis argues that modern humans evolved semi-independently in Europe, Asia and Africa between 100,000 and 40,000 years ago, whereas the 'out of Africa' hypothesis contends that modern humans evolved in Africa between 200 and 100 kyr ago, migrating to Eurasia at some later time. Direct palaeontological, archaeological and biological evidence is necessary to resolve this debate. Here we report the discovery of early Middle Stone Age artefacts in an emerged reef terrace on the Red Sea coast of Eritrea, which we date to the last interglacial (about 125 kyr ago) using U-Th mass spectrometry techniques on fossil corals. The geological setting of these artefacts shows that early humans occupied coastal areas and exploited near-shore marine food resources in East Africa by this time. Together with similar, tentatively dated discoveries from South Africa this is the earliest well-dated evidence for human adaptation to a coastal marine environment, heralding an expansion in the range and complexity of human behaviour from one end of Africa to the other. This new, wide-spread adaptive strategy may, in part, signal the onset of modern human behaviour, which supports an African origin for modern humans by 125 kyr ago. PMID- 10811219 TI - The origin of red algae and the evolution of chloroplasts. AB - Chloroplast structure and genome analyses support the hypothesis that three groups of organisms originated from the primary photosynthetic endosymbiosis between a cyanobacterium and a eukaryotic host: green plants (green algae + land plants), red algae and glaucophytes (for example, Cyanophora). Although phylogenies based on several mitochondrial genes support a specific green plants/red algae relationship, the phylogenetic analysis of nucleus-encoded genes yields inconclusive, sometimes contradictory results. To address this problem, we have analysed an alternative nuclear marker, elongation factor 2, and included new red algae and protist sequences. Here we provide significant support for a sisterhood of green plants and red algae. This sisterhood is also significantly supported by a multi-gene analysis of a fusion of 13 nuclear markers (5,171 amino acids). In addition, the analysis of an alternative fusion of 6 nuclear markers (1,938 amino acids) indicates that glaucophytes may be the closest relatives to the green plants/red algae group. Thus, our study provides evidence from nuclear markers for a single primary endosymbiosis at the origin of these groups, and supports a kingdom Plantae comprising green plants, red algae and glaucophytes. PMID- 10811220 TI - Encoding of movement time by populations of cerebellar Purkinje cells. AB - One of the earliest computational principles attributed to the cerebellum was the measurement of time. This idea was originally suggested on anatomical grounds, and was taken up again to explain some of the deficits in cerebellar patients. The contribution of the cerebellum to eye movements, in contrast, has traditionally been discussed in the context of motor learning. This view has received support from the loss of saccade adaptation, one of the key examples of motor learning, following lesions of the posterior cerebellar vermis. However, the relationship between the properties of saccade-related vermal Purkinje cells and the behavioural deficits that follow lesions is unclear. Here we report results from single-unit recording experiments on monkeys that reconcile the seemingly unrelated concepts of timing and motor learning. We report that, unlike individual Purkinje cells, the population response of larger groups of Purkinje cells gives a precise temporal signature of saccade onset and offset. Thus a vermal population response may help to determine saccade duration. Modifying the time course of the population response by changing the weights of the contributing individual Purkinje cells, discharging at different times relative to the saccade, would directly translate into changes in saccade amplitude. PMID- 10811221 TI - Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation. AB - Vertebrate gastrulation involves the specification and coordinated movement of large populations of cells that give rise to the ectodermal, mesodermal and endodermal germ layers. Although many of the genes involved in the specification of cell identity during this process have been identified, little is known of the genes that coordinate cell movement. Here we show that the zebrafish silberblick (slb) locus encodes Wnt11 and that Slb/Wnt11 activity is required for cells to undergo correct convergent extension movements during gastrulation. In the absence of Slb/Wnt11 function, abnormal extension of axial tissue results in cyclopia and other midline defects in the head. The requirement for Slb/Wnt11 is cell non-autonomous, and our results indicate that the correct extension of axial tissue is at least partly dependent on medio-lateral cell intercalation in paraxial tissue. We also show that the slb phenotype is rescued by a truncated form of Dishevelled that does not signal through the canonical Wnt pathway, suggesting that, as in flies, Wnt signalling might mediate morphogenetic events through a divergent signal transduction cascade. Our results provide genetic and experimental evidence that Wnt activity in lateral tissues has a crucial role in driving the convergent extension movements underlying vertebrate gastrulation. PMID- 10811222 TI - Dishevelled controls cell polarity during Xenopus gastrulation. AB - Although cell movements are vital for establishing the normal architecture of embryos, it is unclear how these movements are regulated during development in vertebrates. Inhibition of Xenopus Dishevelled (Xdsh) function disrupts convergent extension movements of cells during gastrulation, but the mechanism of this effect is unclear, as cell fates are not affected. In Drosophila, Dishevelled controls both cell fate and cell polarity, but whether Dishevelled is involved in controlling cell polarity in vertebrate embryos has not been investigated. Here we show, using time-lapse confocal microscopy, that the failure of cells lacking Xdsh function to undergo convergent extension results from defects in cell polarity. Furthermore, Xdsh mutations that inhibit convergent extension correspond to mutations in Drosophila Dishevelled that selectively perturb planar cell polarity. Finally, the localization of Xdsh at the membrane of normal dorsal mesodermal cells is consistent with Xdsh controlling cell polarity. Our results show that polarized cell behaviour is essential for convergent extension and is controlled by vertebrate Dishevelled. Thus, a vertebrate equivalent of the Drosophila planar cell polarity signalling cascade may be required for normal gastrulation. PMID- 10811223 TI - A receptor for phosphatidylserine-specific clearance of apoptotic cells. AB - cytosis of cellular corpses. During apoptosis, the asymmetry of plasma membrane phospholipids is lost, which exposes phosphatidylserine externally. The phagocytosis of apoptotic cells can be inhibited stereospecifically by phosphatidylserine and its structural analogues, but not by other anionic phospholipids, suggesting that phosphatidylserine is specifically recognized. Using phage display, we have cloned a gene that appears to recognize phosphatidylserine on apoptotic cells. Here we show that this gene, when transfected into B and T lymphocytes, enables them to recognize and engulf apoptotic cells in a phosphatidylserine-specific manner. Flow cytometric analysis using a monoclonal antibody suggested that the protein is expressed on the surface of macrophages, fibroblasts and epithelial cells; this antibody, like phosphatidylserine liposomes, inhibited the phagocytosis of apoptotic cells and, in macrophages, induced an anti-inflammatory state. This candidate phosphatidylserine receptor is highly homologous to genes of unknown function in Caenorhabditis elegans and Drosophila melanogaster, suggesting that phosphatidylserine recognition on apoptotic cells during their removal by phagocytes is highly conserved throughout phylogeny. PMID- 10811224 TI - JNK is required for effector T-cell function but not for T-cell activation. AB - The hallmark of T-cell activation is the production of interleukin 2 (IL-2). c Jun amino-terminal kinase (JNK), a MAP kinase that phosphorylates c-Jun and other components of the AP-1 group of transcription factors, has been implicated in the activation of IL-2 expression. Previously, we found that T cells from mice deficient in the Jnk1 or Jnk2 gene can be activated and produce IL-2 normally, but are deficient in functional differentiation into Th1 or Th2 subsets. However, studies of mice with compound mutations indicate that JNK1 and JNK2 are redundant during mouse development. Here we use three new mouse models in which peripheral T cells completely lack JNK proteins or signalling, to test whether the JNK signalling pathway is crucial for IL-2 expression and T-cell activation. Unexpectedly, these T cells made more IL-2 and proliferated better than wild-type cells. However, production of effector T-cell cytokines did require JNK. Thus, JNK is necessary for T-cell differentiation but not for naive T-cell activation. PMID- 10811225 TI - A role for excreted quinones in extracellular electron transfer. AB - Respiratory processes in bacteria are remarkable because of their ability to use a variety of compounds, including insoluble minerals, as terminal electron acceptors. Although much is known about microbial electron transport to soluble electron acceptors, little is understood about electron transport to insoluble compounds such as ferric oxides. In anaerobic environments, humic substances can serve as electron acceptors and also as electron shuttles to ferric oxides. To explore this process, we identified mutants in Shewanella putrefaciens that are unable to respire on humic substances. Here we show that these mutants contain disruptions in a gene that is involved in the biosynthesis of menaquinone. During growth, the wild type releases a menaquinone-related redox-active small molecule into the medium that complements the mutants. This finding raises the possibility that electron transfer to a variety of oxidants, including poorly soluble minerals, may be mediated by microbially excreted quinones that have yet to be identified. PMID- 10811226 TI - Structural insights into the stereochemistry of the cyclooxygenase reaction. AB - Cyclooxygenases are bifunctional enzymes that catalyse the first committed step in the synthesis of prostaglandins, thromboxanes and other eicosanoids. The two known cyclooxygenases isoforms share a high degree of amino-acid sequence similarity, structural topology and an identical catalytic mechanism. Cyclooxygenase enzymes catalyse two sequential reactions in spatially distinct, but mechanistically coupled active sites. The initial cyclooxygenase reaction converts arachidonic acid (which is achiral) to prostaglandin G2 (which has five chiral centres). The subsequent peroxidase reaction reduces prostaglandin G2 to prostaglandin H2. Here we report the co-crystal structures of murine apo cyclooxygenase-2 in complex with arachidonic acid and prostaglandin. These structures suggest the molecular basis for the stereospecificity of prostaglandin G2 synthesis. PMID- 10811227 TI - Molecular engineering of matrix-targeted retroviral vectors incorporating a surveillance function inherent in von Willebrand factor. AB - A major obstacle that limits the potential of human gene therapy is the inefficiency of gene delivery to appropriate sites in vivo. Previous studies demonstrated that the physiological surveillance function performed by von Willebrand factor (vWF) could be incorporated into retroviral vectors by molecular engineering of the MuLV ecotropic envelope (Env) protein. To advance the application of vWF targeting technology beyond laboratory animals, we prepared an extensive series of Env proteins bearing modified vWF-derived matrix binding sequences and assembled these chimeric proteins into targeted vectors that are capable of transducing human cells. Initially, a dual envelope configuration was utilized, which required coexpression of a wild-type amphotropic Env. Subsequently, streamlined "escort" Env proteins were constructed wherein the inoperative receptor-binding domain of the targeting partner was replaced by the vWF-derived collagen-binding motif. Ultimately, an optimal construct was developed that exhibited properties of both extracellular matrix (ECM)-targeting and near wild-type amphotropic infectivity, and could be arrayed as a single envelope on a retroviral particle. On intraarterial instillation, enhanced focal transduction of neointimal cells (approximately 20%) was demonstrated in a rat model of balloon angioplasty. Moreover, transduction of tumor foci (approximately 1-3%) was detected after portal vein infusion of a matrix-targeted vector in a nude mouse model of liver metastasis. We conclude that the unique properties of these targeted injectable retroviral vectors would be suitable for improving therapeutic gene delivery in numerous clinical applications, including vascular restenosis, laser and other surgical procedures, orthopedic injuries, wound healing, ischemia, arthritis, inflammatory disease, and metastatic cancer. PMID- 10811228 TI - Improved gene transfer to neuroblastoma cells by a monoclonal antibody targeting RET, a receptor tyrosine kinase. AB - Receptor-mediated gene transfer is an effective strategy among nonviral vector systems. It is, however, crucial to develop various types of monoclonal antibodies satisfying both the binding specificity for cell targeting and the capacity of endocytosis required for gene transfer. In the present study, we generated a novel monoclonal antibody (NBL-1) to RET, a receptor tyrosine kinase expressed in both neuroblastoma cells and cells present in substantia nigra, a responsive locus of Parkinson's disease. NBL-1, when added to the culture medium of the neuroblastoma cells, was incorporated by endocytosis in a wortmannin sensitive manner. Using a biotinylated NBL-1 complexed with plasmid DNAs based on electrostatic interaction through avidin-conjugated polylysines, exogenous luciferase genes were expressed in neuroblastoma cells at a more than 10-fold higher level. The expression level of the gene based on NBL-1 was comparable to that obtained by a geneporter system, an improved nonviral gene transduction method. Furthermore, the NBL-1-based gene transfer mediated the formation of more than 20-fold higher numbers of drug-resistant colonies. In contrast, RET-negative cells, which included HeLa, HT1080, Caco-2, and Colo205 cells, did not show any increased expression of an exogenous gene by NBL-1. These data suggest that the RET molecules enable selective gene transduction, and that NBL-1 may possibly be applied to gene therapy for neuroblastomas and Parkinson's disease. PMID- 10811229 TI - Efficient gene transfer into primary human CD8+ T lymphocytes by MuLV-10A1 retrovirus pseudotype. AB - Efficient and stable gene transfer into primary human T lymphocytes would greatly improve their use for adoptive transfer to treat acquired disorders, viral diseases, and cancer. We have constructed retroviral vector pseudotypes of amphotropic murine leukemia viruses (A-MuLV, MuLV-10A1), gibbon ape leukemia virus (GaLV), and feline endogenous virus (RD114) containing the enhanced green fluorescent protein (GFP) as a marker gene. Transduction of primary human CD8+ T lymphocytes by the different GFP-retrovirus pseudotypes revealed the superiority of MuLV-10A1 in comparison with A-MuLV, GaLV, and RD114, respectively. The superior transduction efficacy of CD8+ T cells by MuLV-10A1 correlates with a longer half-life of this pseudotype in comparison with A-MuLV and, as shown by interference analysis with the human T cell line HUT78, by the utilization of both the A-MuLV receptor (Pit2) and the GaLV receptor (Pit1) for cell entry. PMID- 10811230 TI - Highly efficient adenovirus-mediated gene transfer to cardiac myocytes after single-pass coronary delivery. AB - Efficient and homogeneous gene transfer to cardiac myocytes is a major target in myocardial gene therapy. The aim of this study was to determine the conditions permitting efficient, homogeneous, adenovirus-mediated gene transfer to cardiac myocytes, with a view to application during coronary artery catheterization. Gene transfer to adult rat ventricular myocytes was conducted using type 5 adenoviruses carrying the lacZ reporter gene. Adenovirus delivery via coronary arteries was performed on isolated perfused rat hearts, and gene transfer efficiency was analyzed on whole ventricles, freshly isolated myocytes, and cultured myocytes. Single-pass delivery of 1 X 10(9) PFU associated with 1 min of no-flow yielded only 1 +/- 0.5% of positive myocytes. Pretreatment by histamine perfusion (10(-5) M final concentration) increased this value to 30 +/- 9% (p < 0.001), and pretreatment by Ca2+-free buffer perfusion increased it to 67 +/- 8% (p < 0.001). Combination of the two pretreatments had no additional effect. Increasing the viral dose to 3 X 10(9) PFU increased transfection efficiency only in permeabilized vessels. The 1-min no-flow period after adenovirus delivery was crucial for efficient gene transfer: despite histamine pretreatment, only 2 +/- 1% positive myocytes were observed without flow interruption (p < 0.05 versus 1 min of no-flow). Gene transfer was shown to occur in situ during cardiac perfusion, rather than during heart digestion or myocyte isolation. This study shows that highly efficient adenovirus-mediated gene transfer to cardiac myocytes in situ can be achieved by single-pass intracoronary vector delivery, provided that vascular permeability is first increased and coronary flow is briefly interrupted. PMID- 10811231 TI - Transfection of nasal mucosa with a normal alpha1-antitrypsin gene in alpha1 antitrypsin-deficient subjects: comparison with protein therapy. AB - We sought to determine whether a normal alpha1-antitrypsin (AAT) gene could be expressed in respiratory epithelium and whether local expression would have antiinflammatory effects. In an unblinded study, we delivered a normal AAT gene in a plasmid-cationic liposome complex to one nostril of each of five subjects with AAT deficiency; the other, untreated nostril served as a control. AAT protein concentration in nasal lavage fluid (NALF) increased in the transfected nostril (TN), but not in the control nostril (CN), of every subject, peaking on day 5 at levels about one-third normal (baseline CN, 4.1 +/- 1.2 microg/mg of protein; baseline TN, 4.3 +/- 1.3; day 5 CN, 4.0 +/- 0.5 [p = NS versus baseline]; day 5 TN, 9.0 +/- 1.7 [p < 0.5 versus baseline]); isoelectric focusing identified the transgene-generated protein (M) in the only two patients in whom the measurement was possible. The reverse transcriptase-polymerase chain reaction (RT-PCR), performed on NALF from TN and CN of four of the five subjects, was positive for transgene message in TN in all cases and negative in NALF from CN except for one time point in one subject. Interleukin 8 (IL-8) concentrations in NALF were elevated at baseline (normal [N = 10] = 2.5 +/- 0.5 ng/mg of protein; baseline TN = 5.5 +/- 0.8, p < 0.05 versus normal) and decreased after AAT transfection (TN = 2.9 +/- 0.6, p < 0.05 versus baseline) but not in the control nostril (CN = 6.5 +/- 2.2, p = NS versus baseline). NALF samples taken from four of the patients while receiving intravenous AAT protein showed normal concentrations of AAT, but IL-8 concentrations (10.5 +/- 4.2 ng/mg of protein, p = NS versus baseline) were not decreased from baseline. We conclude that plasmid cationic liposome delivery of a normal AAT gene to the respiratory epithelium of deficient patients produces potentially therapeutic local AAT concentrations and that AAT gene therapy, unlike AAT protein therapy, is antiinflammatory. PMID- 10811232 TI - Inhibition of Fas-mediated apoptosis in mouse insulinoma betaTC-3 cells via an anti-Fas ribozyme. AB - In this study we have designed and constructed an anti-Fas ribozyme and show that it can specifically cleave the Fas mRNA in vitro. Moreover, to test its efficacy ex vivo, we transfected the anti-Fas ribozyme into betaTC-3 insulinoma cells, using a RNA polymerase III promoter to drive its expression. Like pancreatic beta cells, betaTC-3 cells do not constitutively express Fas, but Fas expression can be induced with IL-1 and IFN-gamma. Transfected cells expressed an average of 5000 copies of anti-Fas ribozyme transcript per cell as assessed by reverse transcriptase-real-time PCR. After IL-1/IFN-gamma treatment, betaTC-3 cells transfected with the anti-Fas ribozyme expressed 80% less Fas compared with mock transfected cells. In addition, the anti-Fas ribozyme also inhibited Fas expression in NIT-1 insulinoma cells and in primary cultures of dispersed pancreatic islet cells. Inhibition of de novo Fas expression in betaTC-3 cells expressing the anti-Fas ribozyme correlated with resistance to Fas-mediated apoptosis as determined by the number of cells exhibiting caspase 3 proteolytic activity. Hence, we have engineered a ribozyme capable of preventing Fas expression in the betaTC-3 pancreatic insulinoma cell line and conferring resistance to Fas-mediated apoptosis. We suggest that ribozymes may be potentially useful to engineer resistance to apoptosis in transplantable beta cells, a feature that may significantly improve the survival of islet cell grafts. PMID- 10811233 TI - Immunomodulation and adenoviral gene transfer to the lungs of nonhuman primates. AB - Previous data from our laboratory and others have demonstrated a critical role for the CD4+ T lymphocyte in in vivo immune responses to recombinant adenoviral vectors. In rodent models, this subset of T cells is required for T cell proliferation, subsequent cytotoxic T cell generation, and production of anti adenoviral antibodies by B cells. Both depleting and nondepleting anti-CD4 antibodies can attenuate these immune responses to recombinant adenovirus. On the basis of these data, we hypothesized that a nondepleting CDR-engrafted anti-human CD4 antibody (OKT4A) with cross-reactivity to rhesus macaques would attenuate both T and B cell responses to intrapulmonary administration of recombinant adenovirus and permit prolonged reporter gene expression and permit secondary gene transfer. Juvenile rhesus macaques were treated with PBS or OKT4A antibody (10 mg/kg) daily beginning 1 day prior to and up to 11 days after gene transfer. OKT4A resulted in significant attenuation of lymphocyte recruitment into the lung, lymphocyte-proliferative responses to both adenovirus capsid proteins and transgene protein, and adenovirus-induced interferon-gamma elaboration in whole blood and hilar lymph nodes. However, OKT4A was ineffective in attenuating adenovirus-induced IL-4 production in whole blood or hilar lymph nodes, generating neutralizing anti-adenoviral antibodies, or permitting secondary gene transfer. As all the monkeys in this protocol had baseline-detectable anti adenoviral antibodies by ELISA that were nonneutralizing, analogous to most patients with cystic fibrosis, we postulate that anti-CD4 did not block the proliferation of memory B cells. Moreover, these data suggest that for transient immunomodulation to be successful, strategies need to focus specifically on B cell activation independent of CD4+ T cell help. PMID- 10811234 TI - Limitations of informed consent for in utero gene transfer research: implications for investigators and institutional review boards. AB - Only 10 years after the first human gene transfer protocols were approved for adults and children, researchers have begun to consider gene transfer on the fetus. While preliminary animal research is ongoing, the enthusiasm and pace of research in this area suggest that human protocols for in utero gene transfer research may be seriously considered in the foreseeable future. Federal guidelines for fetal research rely on minimizing risk and informed consent to protect the "rights and welfare" of both the fetus and pregnant woman. However, in utero gene transfer research poses special challenges to informed consent. This research represents an innovative approach for very ill subjects and takes place in the prenatal setting. These features may converge to undermine the expectant parents' comprehension of, and voluntariness for participation in, research. In this case, informed consent may not be able to bear the weight of adequately protecting the fetus from undue research risks. To compensate for this limitation, and using the regulations for pediatric research as a guide, a greater emphasis should be placed on the benefit/harm assessment rather than informed consent. Selecting diseases/patients where good alternative treatments exist may maximize informed consent, yet this may be a trade-off that exposes the fetus to greater relative risks. On the other hand, selecting diseases/patients without good alternative treatments to prolong life may convey an overestimation of the potential benefits of these interventions, and although care should be taken to strive to improve understanding of these limitations, misunderstanding may persist. However, selecting diseases/patients with no good alternatives might make serious risks more tolerable, and this should take precedence over informed consent. The limitations of informed consent brought into focus by the special features of in utero gene transfer research may be relevant to a broader range of innovative investigations. PMID- 10811235 TI - A phase I trial of intra lesional RV-B7.1 vaccine in the treatment of malignant melanoma. PMID- 10811236 TI - Side Effects of Antipsychotic Medications: Physician's Choice of Medication and Patient Compliance. Dallas, Texas, California. January 22, 1999. Proceedings of a roundtable. PMID- 10811237 TI - An overview of side effects caused by typical antipsychotics. AB - Typical antipsychotics often combine efficacy in treating antipsychotic illnesses with a side effect profile that can affect every system of the body and range from the annoying-photosensitivity and jaundice, for example-to the disabling seizures and blindness, among others-to the potentially fatal-agranulocytosis and neuroleptic malignant syndrome. The side effects of conventional antipsychotics are associated with effects at CNS transmission and receptor sites and appear in relation to dose levels and potency of the drug. Characteristics of patients including gender, age, and comorbid medical illness-can make them more or less susceptible to particular antipsychotic side effects. Side effects influence patient quality of life and affect patient compliance with medications. This article will consider the physiologic systems affected by conventional neuroleptics, the sexual and reproductive side effects of typical antipsychotics, and the central nervous side effects of the conventional neuroleptics in the light of these concerns. PMID- 10811238 TI - Review and management of clozapine side effects. AB - Clozapine has demonstrated superior efficacy in relieving positive and negative symptoms in treatment-resistant schizophrenic patients; unlike other antipsychotics, it causes minimal extrapyramidal side effects (EPS) and has little effect on serum prolactin. Despite these benefits, the use of clozapine has been limited because of infrequent but serious side effects, the most notable being agranulocytosis. In recent years, however, mandatory blood monitoring has significantly reduced both the incidence of agranulocytosis and its associated mortality. The occurrence of seizures appears to be dose-related and can generally be managed by reduction in clozapine dosage. Less serious and more common side effects of clozapine including sedation, hypersalivation, tachycardia, hypotension, hypertension, weight gain, constipation, urinary incontinence, and fever can often be managed medically and are generally tolerated by the patient. Appropriate management of clozapine side effects facilitates a maximization of the benefits of clozapine treatment, and physicians and patients alike should be aware that there is a range of benefits to clozapine use that is wider than its risks. PMID- 10811239 TI - Risperidone side effects. AB - The overall effectiveness of traditional antipsychotics has been hindered by their extrapyramidal side effects, which contribute to noncompliance and relapse in patients with schizophrenia. The side effects associated with traditional antipsychotic treatment are generally minimal in patients who take risperidone, a combined 5-HT2/D2 antagonist, but the literature is sparse on adverse events among the newer atypical antipsychotics. Risperidone is associated with relatively few motor side effects compared with the traditional antipsychotics, and weight gain is less likely with risperidone than with either clozapine or olanzapine. While increased prolactin levels have been reported in patients taking risperidone, little correlation has been found between prolactin levels and adverse events. As antipsychotic treatment options expand to include the new agents, it is important for clinicians to anticipate side effects and to query patients about specific adverse events. PMID- 10811240 TI - Adverse events related to olanzapine. AB - Olanzapine, a serotonin-dopamine receptor antagonist, is one of the novel atypical antipsychotics that is effective against the positive and negative symptoms of schizophrenia with significantly fewer treatment-emergent extrapyramidal symptoms and less akathisia associated with traditional antipsychotics. Compared with traditional agents, olanzapine shows only a few adverse events such as dry mouth, sedation, and increase in appetite. Compared with risperidone, olanzapine causes greater increases in weight gain and body mass index but less hyperprolactinemia. Transient, non-dose-dependent, asymptomatic elevations in liver enzymes have also been noted in olanzapine treated patients. Because of the comparative efficacy and improved side effect profiles of the atypical antipsychotics, consideration should be given to using the newer agents as preferred treatment for schizophrenia and related psychoses. PMID- 10811241 TI - Review of quetiapine side effects. AB - The newest atypical antipsychotic medication to be approved by the U.S. Food and Drug Administration, quetiapine is a drug that awaits a wide range of clinical and head-to-head comparisons. Nevertheless, clinical trials currently available suggest that quetiapine has a beneficial side effect profile, particularly with regard,to extrapyramidal symptoms. To date, quetiapine has also proved effective in the treatment of schizophrenia, but its efficacy, while clearly superior to that of placebo, seems no greater than that of haloperidol or chlorpromazine. Clinical trials have supported the use of quetiapine in treating elderly patients. Further research is necessary to establish the clinical profile of quetiapine. PMID- 10811242 TI - Weight gain and antipsychotic medication. AB - Antipsychotic-treated patients, who tend to take combinations of psychotropic agents that may cause weight gain, are at special risk for the problems associated with being overweight or obese. Medical and psychiatric examinations should include periodic monitoring for weight change and an obesity assessment for weight-related medical illness. The assessment should involve evaluation of body mass index and waist circumference as well as medical history. Preventative strategies should be undertaken for patients who gain 5 lb (2.3 kg) or more within a 3-year period. A change in antipsychotics may be necessary for overweight patients who are unwilling or unable to lose weight. A treatment plan for overweight or obese patients should include periodic monitoring and recommendations for changes in diet and physical activity. Support groups and adjunct medication may also be helpful. The patient should be reminded of the benefits of even modest weight loss. PMID- 10811243 TI - Side effects of antipsychotics in the elderly. AB - Side effects of antipsychotic medications are particularly problematic in elderly patients, who experience many age-related changes that may exacerbate medication side effects. Side effects of particular concern in the elderly include anticholinergic reactions, parkinsonian events, tardive dyskinesia, orthostatic hypotension, cardiac conduction disturbances, reduced bone mineral density, sedation, and cognitive slowing. In addition, elderly patients with schizophrenia often have comorbid medical illnesses-such as cardiovascular disease and dementia of the Alzheimer's type-and are thus likely to be taking multiple medications. The effects of polypharmacy must be carefully considered. Patients, caregivers, and family often have different perspectives on side effects. This article addresses the side effects of the currently available antipsychotic medications in light of these concerns. PMID- 10811244 TI - Antipsychotic drug side effect issues in bipolar manic patients. AB - While the efficacy of antipsychotics as a maintenance treatment for bipolar patients has not been systematically studied, these drugs are commonly used in the long-term treatment of bipolar patients, and it is not unusual for a bipolar patient to be taking 3 to 4 medications, including antipsychotics. Conventional antipsychotics may be comparable to lithium for acute mania, but have limitations when used in the long-term treatment of bipolar disorder. Their adverse effects, which include extrapyramidal side effects, tardive dyskinesia, weight gain, sedation, and sexual dysfunction, often lead to non-compliance; their use may have a negative impact on the overall course of illness; and they may not be as effective as lithium in treating the core manic symptoms over the long term. Atypical antipsychotics may prove useful for bipolar patients who require antipsychotic treatment because of their favorable side effect profile, thymoleptic properties, and positive effect on overall functioning. PMID- 10811246 TI - Reproductive outcomes in a population exposed long-term to inorganic selenium via drinking water. AB - Despite being an essential element in mammals and lower animals at very low doses, selenium is recognized as a teratogen and a growth-inhibiting substance in several animal species. Some inorganic selenium compounds have also been shown to be mutagenic and pro-oxidant: however, very little is known about their effects on human reproduction. Between 1972 and 1988, a few thousand residents in the municipality of Reggio Emilia, northern Italy, were accidentally exposed to drinking water with unusually high levels of inorganic selenium (selenate), range 7-9 microg/l, through a local public water supply system. We found no deleterious effect on overall body weight and length of newborns and stillborns delivered by 18 women previously exposed to high-selenium tapwater. Rates of spontaneous abortions, however, were increased slightly (RR = 1.73; 95% CI = 0.62-4.80), compared with rates among unexposed women from the same municipality. From 1980 through 1988 the prevalence of congenital malformations at birth among the exposed population was similar to that expected, but estimates were imprecise owing to small numbers. Results do not suggest marked effects on human reproduction from chronic exposure to selenate in drinking water at < 10 microg/l. PMID- 10811247 TI - Total metal concentrations and partitioning of Cd, Cr, Cu, Fe, Ni and Zn in sewage sludge. AB - Application of the BCR three-step sequential extraction procedure to sewage sludge samples collected at an urban wastewater treatment plant (Domzale, Slovenia) is reported. The total concentrations of Cd, Cr, Cu, Fe, Ni and Zn and their concentrations in fractions after extraction were determined by flame or electrothermal atomic absorption spectrometry (FAAS, ETAAS) under optimised measurement conditions. Total acid digestion including hydrofluoric acid (HF) treatment and aqua regia extraction were compared in order to estimate the efficiency of aqua regia extraction for determination of total metal concentrations in sewage sludge. It was found experimentally that aqua regia quantitatively leached these heavy metals from the sewage sludge and could therefore be applied in analysis of total heavy metal concentrations. The total concentrations of 856 mg kg(-1) Cr, 621 mg kg(-1) Ni and 2032 mg kg(-1) Zn were higher than those set by Slovenian legislation for sludge to be used in agriculture. Total concentrations of 2.78 mg kg(-1) Cd, 433 mg kg(-1) Cu and 126 mg kg(-1) Pb were below those permitted in the relevant legislation. CRM 146R reference material was used to follow the quality of the analytical process. The results of the BCR three-step sequential extraction procedure indicate high Ni and Zn mobility in the sludge analysed. The other heavy metals were primarily in sparingly soluble fractions and hence poorly mobile. Due to the high total Ni concentration and its high mobility the investigated sewage sludge could not be used in agriculture. PMID- 10811248 TI - Biological reduction of uranium in groundwater and subsurface soil. AB - Biological reduction of uranium is one of the techniques currently studied for in situ remediation of groundwater and subsurface soil. We investigated U(VI) reduction in groundwaters and soils of different origin to verify the presence of bacteria capable of U(VI) reduction. The groundwaters originated from mill tailings sites with U concentrations as high as 50 mg/l, and from other sites where uranium is not a contaminant, but was added in the laboratory to reach concentrations up to 11 mg/l. All waters contained nitrate and sulfate. After oxygen and nitrate reduction, U(VI) was reduced by sulfate-reducing bacteria, whose growth was stimulated by ethanol and trimetaphosphate. Uranium precipitated as hydrated uraninite (UO2 x xH2O). In the course of reduction of U(VI), Mn(IV) and Fe(III) from the soil were reduced as well. During uraninite precipitation a comparatively large mass of iron sulfides formed and served as a redox buffer. If the excess of iron sulfide is large enough, uraninite will not be oxidized by oxygenated groundwater. We show that bacteria capable of reducing U(VI) to U(IV) are ubiquitous in nature. The uranium reducers are primarily sulfate reducers and are stimulated by adding nutrients to the groundwater. PMID- 10811249 TI - Emissions of lead and zinc from candles with metal-core wicks. AB - We measured the amount of lead released from 14 different brands of candles with metal-core wicks sold in Michigan. The emissions of lead were found to range from 0.5 to 66 microg/h, and the rates for zinc were from 1.2 to 124 microg/h. It is estimated that burning four of the candles bought in Michigan for 2 h can result in airborne lead concentrations that can pose a threat to human health. In addition to inhalation of lead in the air, children get exposed to lead in candle fumes deposited on the floor, furniture and walls through their hand-to-mouth activity. Burning candles with leaded-core wick may be an important exposure route for lead that has generally been ignored. PMID- 10811250 TI - Daily intake of manganese by the adult population of Mumbai. AB - The daily intake of manganese (Mn) estimated through air, water and duplicate dietary analysis is found to range from 0.67 to 4.99 mg with a mean value of 2.21 mg. Ingestion through food contributed to the predominant fraction of the intake. The turnover rate of Mn through blood is approximately 2 h, based on the mean concentration of Mn in blood of 1.54 microg l(-1). The average concentrations of Mn in water and air were approximately 1.42 microg l(-1) and 37 ng m(-3), respectively. The daily intake of Mn by the adult population of Mumbai is closer to the lower bound of the recommended limit of 2-5 mg. Electro Thermal Atomic Absorption Spectrophotometry (ET-AAS), has been used for the determination of Mn in a variety of environmental and human biological fluids. The detection limit of Mn for a volume injection of 20 microl is 2 pg absolute. The precision of the method is established by analyzing a synthetic mixture containing various elements in different quantities (0.5-10 ppm) and is found to be within +/- 8%. The reliability of estimation is further assessed through the analysis of Standard Reference Materials (SRMs) of soil, hay, milk powder and fish tissue obtained from IAEA. PMID- 10811251 TI - Sources and concentrations of indoor nitrogen dioxide in Hamburg (west Germany) and Erfurt (east Germany). AB - Here we report indoor and outdoor concentrations of NO2 for Erfurt and Hamburg and assess the contribution of the most important indoor sources (e.g. the presence of gas cooking ranges, smoking) and outdoor sources (traffic exhaust emissions). We examined the relative contribution of the different sources of NO2 to the total indoor NO2 levels in Erfurt and Hamburg. NO2 indoor concentrations in Hamburg were slightly higher than those in Erfurt (i.e. living room: 15 microg m(-3) for Erfurt and 17 microg m(-3) for Hamburg). A linear regression model including the variables, place of residence, season and outdoor NO2 levels, location of the home within the city, housing and occupant characteristics accounted for 38% of the NO2 variance. The most important predictors of indoor NO2 concentrations were gas in cooking followed by other characteristics, such as ventilation or outdoor NO2 level. Residences in which gas was used for cooking, or in which occupants smoked, had substantially higher indoor NO2 concentrations (41 or 18% increase, respectively). An increase in the outdoor NO2 concentration from the 25th to the 75th-percentile (17 microg m(-3)) was associated with a 33% increase in the living room NO2 concentration. Multiple regression analysis for both cities separately illustrated that use of gas for cooking was the major indoor source of NO2. This variable caused a similar increase in the indoor NO2 levels in each city (43% in Erfurt and 47% in Hamburg). However, outdoor sources of NO2 (motor vehicle traffic) contributed more to indoor NO2 levels in Hamburg than in Erfurt. PMID- 10811252 TI - The kinetics and reversibility of the partitioning of polychlorinated biphenyls between air and ryegrass. AB - Lolium multiflorum (ryegrass) was contaminated with technical mixtures of polychlorinated biphenyls (PCBs) via the gas phase and the subsequent elimination of the PCBs from the vegetation was studied under field conditions. There was a pronounced decrease in the concentrations of the di- through to the pentachlorinated congeners over the 240-h elimination period. For many of the congeners the elimination was nearly complete, suggesting that the partitioning of these compounds from the gas phase into ryegrass is largely reversible. The elimination followed first order kinetics. The elimination half-lives were linearly proportional to the plant/air equilibrium partition coefficients (K(PA)) of the PCB congeners and ranged from 22 h for PCBs 8 + 5 to 87 h for PCBs 84 + 101. For the hexa- through to the octachlorinated congeners no significant decrease in the concentration of the vegetation was observed during the elimination experiment. The elimination was described well using a two-resistance model of PCB desorption which indicated that elimination of the di- and trichlorinated PCBs was limited by transport within the plant itself while elimination of the higher chlorinated congeners was limited by transport from the plant surface into the atmosphere. Furthermore, the results demonstrate that if elimination of higher chlorinated PCBs from ryegrass via biological or photochemical degradation occurs, then it is very slow. PMID- 10811253 TI - Phosphine generation by mixed- and monoseptic-cultures of anaerobic bacteria. AB - A microbial basis for bioreductive generation of phosphine is proposed, which could account at least in part for the presence of this toxic gas in natural anaerobic environments and in sewage and landfill gases. Phosphine generation under anaerobic growth conditions was dependent upon both the culture inoculum source (animal faeces) and enrichment culture conditions. Phosphine was detected in headspace gases from mixed cultures under conditions promoting fermentative growth of mixed acid and butyric acid bacteria, either in the presence or absence of methane generation. Monoseptic cultures of certain mixed acid fermentors (Escherichia coli, Salmonella gallinarum, and Salmonella arizonae) and solvent fermentors (Clostridium sporogenes, Clostridium acetobutyricum and Clostridium cochliarium) also generated phosphine. Such fermentative bacteria participate in the multi-stage process of methanogenesis in nature. Generation of phosphine by these bacteria, rather than by methanoarchaea themselves, could explain the apparent correlation between methanogenesis and the formation of phosphine in nature. PMID- 10811254 TI - Application of double focusing sector field ICP-MS for multielemental characterization of human hair and nails. Part I. Analytical methodology. AB - The capabilities of double focusing sector field inductively coupled plasma mass spectrometry (ICP-SMS) for the determination of 71 elements in hair and nails were studied. A microwave-assisted digestion procedure with nitric acid and hydrogen peroxide as well as direct sampling of the nails by laser ablation (LA) have been tested. Examples of spectral interferences are given and different correction procedures are discussed. Method detection limits below ng g(-1) were obtained for 39 elements investigated by using high-purity reagents and by taking special care to prevent contamination during preparation. However, these detection limits were insufficient for detection of some platinum group elements in the majority of the samples. The accuracy of the analytical procedure was estimated by analysis of the GBW07601 certified reference material as well as by participation in an interlaboratory comparison program. The reproducibility was assessed from replicate analysis (including sample preparation) and was found to be, as average values for all elements, 9-10% R.S.D. and 18-19% R.S.D. for hair and nails, respectively. Contribution from exogenous deposition was evaluated by analyzing samples before and after washing, as well as by studying spatial element distribution along hair and nails. It was found that even after sample washing, many elements are enriched in the surface of the nail. PMID- 10811255 TI - Long-term processes in waste deposits. AB - A conceptual model, which is a unitary and continuous description of the overall processes in waste deposits, has been developed. In the model the most important processes governing the long-term fate of organic matter in landfills and the transport and retention of toxic metals are included. With the model as a base, a number of scenarios with different levels of complexity have been defined and studied in order to carry out long-term assessments of the chemical evolution in waste deposits for industrial and municipal solid waste containing much organic matter and the leaching of toxic metals. The focus of the modelling has been to quantify the important processes occurring after the methane production phase has ceased, i.e. during the humic phase. The scenarios include the main mechanisms based on various transport processes as well as different landfill constructions, e.g. binding capacities of sulfides and humic substances. They also include transport mechanisms by which the reactant oxygen can intrude into a deposit, sorption capacities of hydrous ferric oxides, and pH-buffering reactions, etc. Scoping calculations have shown that the binding capacity of humic substances is sufficient to bind all toxic metals (Cd, Cr, Pb, Zn and Hg). In addition, the humics could also bind a smaller part of Ca, Fe and Al, provided much of the organic waste remain as humic substances. Sulfides on the other hand can bind approximately twice the amount of all toxic metals. The binding capacity of hydrous ferric oxides, which can be formed by oxidation reactions during the humic phase, is estimated to be three times the total content of metals that can sorb on hydrous ferric oxides. In the studied landfill the pH-buffering capacity, primarily represented by calcite, is estimated to be 1 mol/kg dry waste. Quantifications indicate that the alkalinity of the wastes is high enough to buffer the acidity produced by the oxidation of sulfides and by the degradation of organic matter, as well as that added by acid precipitation. Therefore, the main conclusion is that higher remobilisation rates of heavy metals due to lowering of pH are not expected for many thousands of years. PMID- 10811256 TI - Trace element levels in harp seal (Pagophilus groenlandicus) and hooded seal (Cystophora cristata) from the Greenland Sea. A multivariate approach. AB - Concentrations of the essential trace elements, iron, copper, zinc and selenium and the non-essential elements arsenic, cadmium, total mercury and lead, were measured in the meat, liver and kidney of two species of seals, harp seal (Pagophilus groenlandicus) and hooded seal (Cystophora cristata) collected in the Greenland Sea. The spread among the individual seals was considerable. However, multivariate statistics simplified the evaluation of the data. The muscle tissue contained lower levels of the elements than kidney and liver. The kidney and liver tissue were also different, in particular with higher levels of iron in the liver and higher levels of cadmium in the kidney. Species differences were clear in both liver and kidney tissue, with higher levels of most of the elements in the hooded seals, while the harp seals had a higher burden of arsenic in the two tissues. Male hooded seals had higher levels of mercury and selenium than the females in all tissues. For harp seal there was a slight difference between the sexes in the muscle tissue, while no difference was observed in the liver and kidney tissues. The juvenile seals generally had lower levels of the elements in their tissues than the adults, although copper and zinc were higher in the muscles and livers of the juveniles as was iron in the muscles. No correlation between age and trace element levels in the tissues of the adult seals was observed. PMID- 10811257 TI - Heavy metals in maternal and cord blood. AB - Heavy metal concentrations have been determined in maternal and cord blood samples collected from mothers in the age group 20-25 years with full-term neonates (37-40 weeks). The concentrations of Pb, Cd, Cu and Zn were found to be low in cord blood as compared to mother's blood and the ratio between cord blood and maternal blood for the respective elements was found to be approximately 0.80, 0.86, 0.47 and 0.40. On the other hand, the concentrations of Fe and Mg in the mother's blood were found to be lower than those in the cord blood. Prenatal exposure to lead in Mumbai, though low (5.1 microg/dl), is approximately 2-3 times higher than that observed in Canada or Italy. A strong correlation (r = 0.79) between the maternal and cord blood lead levels has been observed during the present study. PMID- 10811258 TI - A review of water quality concerns in livestock farming areas. AB - Post-war changes in farming systems and especially the move from mixed arable livestock farming towards greater specialisation, together with the general intensification of food production have had adverse affects on the environment. Livestock systems have largely become separated into pasture-based (cattle and sheep) and indoor systems (pigs and poultry). This paper reviews water quality issues in livestock farming areas of the UK. The increased losses of nutrients, farm effluents (particularly livestock wastes), pesticides such as sheep-dipping chemicals, bacterial and protozoan contamination of soil and water are some of the main concerns regarding water quality degradation. There has been a general uncoupling of nutrient cycles, and problems relating to nutrient loss are either short-term direct losses or long-term, related to accumulated nutrient surpluses. Results from several field studies indicate that a rational use of manure and mineral fertilisers can help reduce the pollution problems arising from livestock farming practices. Several best management practices are suggested for the control of nutrient loss and minimising release of pathogen and sheep-dip chemicals into agricultural runoff. PMID- 10811259 TI - Government to review dispensing by veterinary surgeons, together with the use of paraprofessionals. PMID- 10811260 TI - Food Standards Agency gets under way. PMID- 10811261 TI - Of PETS, medicines and disease surveillance. PMID- 10811262 TI - Long-term impact on a closed household of pet cats of natural infection with feline coronavirus, feline leukaemia virus and feline immunodeficiency virus. AB - A closed household of 26 cats in which feline coronavirus (FCoV), feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) were endemic was observed for 10 years. Each cat was seropositive for FCoV on at least one occasion and the infection was maintained by reinfection. After 10 years, three of six surviving cats were still seropositive. Only one cat, which was also infected with FIV, developed feline infectious peritonitis (FIP). Rising anti FCoV antibody titres did not indicate that the cat would develop FIP. The FeLV infection was self-limiting because all seven of the initially viraemic cats died within five years and the remainder were immune. However, FeLV had the greatest impact on mortality. Nine cats were initially FIV-positive and six more cats became infected during the course of the study, without evidence of having been bitten. The FIV infection did not adversely affect the cats' life expectancy. PMID- 10811263 TI - Anatomy of the patent ductus venosus in the dog. AB - The biplanar umbilical vein portovenograms of 49 newborn puppies and the biplanar mesenteric vein portovenograms, obtained during surgery, of 42 adult dogs with left divisional intrahepatic portosystemic shunts consistent with a patent ductus venosus (PDV) were reviewed. On the basis of the combined surgical, postmortem and imaging data, the left divisional intrahepatic portosystemic shunts were consistent, each having a straight vessel which drained into a venous ampulla before draining into the caudal vena cava at the level of the diaphragm. The left phrenic vein and the left hepatic vein both entered the ampulla independently of the shunting vessel. The morphology of the ductus venosus in the pups was similar and consistent with the morphology of the left divisional intrahepatic PDV shunt of the adult dogs. It is concluded that this form of left divisional shunt is correctly named a PDV and is the result of the persistence of the fetal ductus venosus. From the surgical records it is concluded that all the shunts described as a PDV were attenuated by the direct manipulation of the ductus venosus before its entry into the ampulla. PMID- 10811264 TI - Use of a Rush pin to repair fractures of the distal femur in cats. AB - Nine fractures of the distal femur in eight cats were repaired by using a 2.4 mm Rush pin as an intramedullary nail, placed between the condyles via a lateral or medial parapatellar approach to the stifle joint. In all cases the fractures were reduced and stabilised accurately with good postoperative results. PMID- 10811265 TI - Efficacy of oral calcium pentosan polysulphate for the treatment of osteoarthritis of the canine stifle joint secondary to cranial cruciate ligament deficiency. AB - The efficacy of calcium pentosan polysulphate (CaPPS) as a slow-acting drug for the treatment of osteoarthritis of the canine stifle joint, secondary to cranial cruciate ligament deficiency, was tested in a double-blind, placebo-controlled clinical trial over a period of one year. Dogs with the deficiency were treated surgically, matched for bodyweight, and randomly assigned to treatment or placebo groups. Active treatment began six weeks postoperatively and consisted of 10 mg/kg CaPPS orally, once weekly for four weeks, repeated every 12 weeks. The outcome was assessed in terms of function by the dogs' owners, by the radiographical grading of the osteoarthritis, and by the measurement of total sulphated glycosaminoglycans and the 5D4 epitope of keratan sulphate in the synovial fluids of affected joints. There were no differences either in functional outcome or in the radiographical progression of osteoarthritis between the two groups. Fifty-four weeks after surgery, the concentration of 5D4 in synovial fluid (expressed as change from baseline values) had decreased significantly in the treatment group compared with the placebo group (P=0.03). PMID- 10811266 TI - In vitro study of the significance of bladder neck position in incontinent bitches. PMID- 10811267 TI - Temporary lateral tarsorrhaphy for the treatment of lower lateral eyelid entropion in juvenile dogs. PMID- 10811268 TI - Prevalence of dermatophytes in asymptomatic guinea pigs and rabbits. PMID- 10811269 TI - Tendering for LVI work. PMID- 10811270 TI - Tendering for LVI work. PMID- 10811271 TI - Exotic pets. PMID- 10811272 TI - Role of megabacteria in mammals. PMID- 10811273 TI - Plasmodium falciparum sporozoites increase feeding-associated mortality of their mosquito hosts Anopheles gambiae s.l. AB - There is some evidence that pathology induced by heavy malaria infections (many oocysts) increases mortality of infected mosquitoes. However, there is little or no published evidence that documented changes in feeding behaviour associated with malaria infection also contribute to higher mortality of infected mosquitoes relative to uninfected individuals. We show here for the first time that, in a natural situation, infection by the sporozoites of the malaria parasite Plasmodium falciparum significantly reduced survival of blood-feeding Anopheles gambiae, the major vector of malaria in sub-Saharan Africa. To estimate feeding associated mortality of infected mosquitoes, we compared the percentage of sporozoite infection in host-seeking mosquitoes caught before and after feeding. The infection rate was 12% for mosquitoes caught during the night as they were entering a tent to feed; however, only 7.5% of the surviving members of the same cohort caught after they had had the opportunity to feed were infected. Thus, Plasmodium falciparum sporozoites increased the probability of dying during the night-time feeding period by 37.5%. The increase in mortality was probably due to decreased efficiency in obtaining blood and by increased feeding activity of the sporozoite-infected mosquitoes that elicited a greater degree of defensive behaviour of hosts under attack. PMID- 10811274 TI - Genetic restriction of Plasmodium falciparum in an area of stable transmission: an example of island evolution? AB - To date, a high degree of polymorphism has been demonstrated at both the MSP1 and MSP2 loci in parasites from areas of stable malaria transmission. As a consequence, in such areas it is rare to find parasites of the same 2-locus genotype in more than 1 subject. We have studied MSP1 and MSP2 diversity in parasites collected from subjects with both symptomatic (n = 86) and asymptomatic (34) malaria living on the island of Santo, Vanuatu, an area of stable malaria transmission. Polymorphism at the MSP1 and MSP2 loci was considerably less than previously reported: only 5 MSP1 and 5 MSP2 alleles were detected and these showed no size variation within alleles. Santo is unique amongst the areas studied so far in that it is a small island at the limit of the malaria belt in the South Pacific. Thus, the evolution of the parasite population may have been affected by the small size and isolation of this island population. Moreover, limited parasite diversity may explain the unusually mild nature of Plasmodium falciparum disease on Santo. Islands have fascinated biologists for centuries and fuelled the advancement of evolutionary theory, since they are natural laboratories for the study of evolution. The simplicity of the Vanuatu P. falciparum population may facilitate the use and interpretation of sequence level analyses to address the mechanisms by which genetic diversity is generated and maintained in natural populations. PMID- 10811275 TI - Molecular characterization of the ribosomal RNA gene region of Perkinsus atlanticus: its use in phylogenetic analysis and as a target for a molecular diagnosis. AB - Due to their widespread distribution and virulence, protozoan species of the genus Perkinsus are especially worrisome parasites for shellfish farmers. In the present paper, we investigate the organization and the structural features of the nuclear ribosomal genes of Perkinsus atlanticus as well as the use of DNA sequence information from this region for phylogenetic analyses. This information has been useful, further, for the development of a diagnostic test based on the amplification by the polymerase chain reaction (PCR) technique. We have isolated a high-copy DNA sequence in this species, and, after its characterization, we have determined that it corresponds to the ribosomal RNA (rRNA) genes 28S-5S-18S and the intergenic spacers. By comparing the complete sequence of the 5S rRNA gene and a partial sequence of the 18S rRNA gene of P. atlanticus with the sequences of those genes in other Alveolates, we have found additional support for the hypothesis that Perkinsus is more closely related to species of Dinoflagellata than to species of Apicomplexa. The intergenic spacer sequence between the 5S and the 18S rRNA genes was used to design a pair of primers to be used as a PCR-based diagnostic test. PMID- 10811276 TI - Population biology of Eimeria (Protozoa: Apicomplexa) in Apodemus sylvaticus: a capture/recapture study. AB - The first long-term (27 month) survey of single species of Eimeria occurring in a population of Apodemus sylvaticus in the UK showed that Eimeria apionodes, E. hungaryensis and E. uptoni occurred throughout the period whilst E. sp. E. was only found during 4 months. In first-captured animals, overall prevalence of Eimeria spp. was 73%: a figure higher than previously published, but possibly an underestimate. Multiple infections (2 or 3 species) occurred in 34% of first capture infected animals, but without significant associations. There were no significant differences of prevalence rates between the sexes, nor between adults and juveniles, except for E. uptoni which was more common in juveniles than in adults. E. hungaryensis and E. uptoni probably occurred as near-continuous infections whilst E. apionodes occurred more intermittently but with no long-term total immunity. Monthly prevalence data indicated ongoing trends over the 27 months. Prevalences of E. apionodes indicated an annual cycle, lowest in January May and with a peak in October-November, correlating positively with both the number of animals and the percentage of juveniles in the host population and also with the environmental relative humidity. There was no consistent pattern for E. uptoni and an annual cycle for E. hungaryensis was not apparent. PMID- 10811277 TI - 'Babesia gibsoni' of dogs from North America and Asia belong to different species. AB - 18S rDNA sequences from 4 isolates of Babesia gibsoni originating from Japan, Malaysia and Sri Lanka were compared with a previously published, 0.5 kb portion of the 18S rDNA from a B. gibsoni isolate from California, USA, and with the corresponding 18S rDNA sequences of other Babesia spp. Distance, parsimony and maximum likelihood analyses showed almost identical genotypes among the small canine Babesia from Asia, but an unexpectedly distant genetic relationship to that from the USA. While the American isolate segregated together with B. equi, the Asian isolates showed a close relationship to B. divergens and B. odocoilei. These results indicate that small Babesia of dogs originating from North America and Asia belong to different, genetically distantly related species. PMID- 10811278 TI - A statistically derived index for classifying East Coast fever reactions in cattle challenged with Theileria parva under experimental conditions. AB - A statistically derived disease reaction index based on parasitological, clinical and haematological measurements observed in 309 5 to 8-month-old Boran cattle following laboratory challenge with Theileria parva is described. Principal component analysis was applied to 13 measures including first appearance of schizonts, first appearance of piroplasms and first occurrence of pyrexia, together with the duration and severity of these symptoms, and white blood cell count. The first principal component, which was based on approximately equal contributions of the 13 variables, provided the definition for the disease reaction index, defined on a scale of 0-10. As well as providing a more objective measure of the severity of the reaction, the continuous nature of the index score enables more powerful statistical analysis of the data compared with that which has been previously possible through clinically derived categories of non-, mild, moderate and severe reactions. PMID- 10811279 TI - Isolation, characterization and expression of a GRA2 homologue from Neospora caninum. AB - A cDNA library derived from mRNA of tachyzoites of Neospora caninum (NC-Liverpool strain) was screened with antisera from a cow naturally infected with N. caninum. The DNA sequence of 1 recombinant isolated predicted a significant protein sequence homology of the gene product to the 28 kDa (GRA2) antigen of Toxoplasma gondii. Studies on the N. caninum gene coding for this antigen demonstrated the presence of a single intron flanked by 2 exons; the gene was also highly expressed in culture-derived tachyzoites. The antigen was expressed in Escherichia coli; when injected into mice it stimulated the production of antibodies which detected a 29 kDa antigen of N. caninum. Secondary structure predictions made for the N. caninum protein showed support for several amphipathic helices separated by loops and turns. The available evidence indicates maintenance of protein secondary structure, and not DNA or amino acid sequence, has occurred during the evolution of GRA2 proteins in N. caninum and T. gondii. PMID- 10811280 TI - Nitric oxide interaction with IL-10, MIP-1alpha, MCP-1 and RANTES over the in vitro granuloma formation against different Schistosoma mansoni antigenic preparations on human schistosomiasis. AB - Nitric oxide (NO) produced by cytokine-activated macrophages is reported to be cytotoxic against the helminth Schistosoma mansoni, although this is a controversial issue. Previous work in our laboratory identified a fraction of S. mansoni soluble adult worm antigenic preparation (SWAP), named PIII, able to elicit significant in vitro cell proliferation and at the same time lower in vitro and in vivo granuloma formation when compared either to soluble egg antigen (SEA) or to SWAP. Here we report that, in comparison to other S. mansoni antigenic preparations (SEA and SWAP), supernatants of PBMC cultivated with PIII possess higher concentrations of interleukin-10 (IL-10) and macrophage inflammatory protein (MIP-1alpha), concomitantly with lower concentrations of monocyte chemoattractant protein (MCP-1) and regulated on activation, normal T expressed and secreted (RANTES). In the particular case of NO inhibition, supernatants of PBMC cultivated with PIII present decreased IL-10 levels. Altogether, these results indicate that IL-10, MIP-1alpha, MCP-1 and RANTES are distinctively important elements in the PIII modulating role, while NO seems to be pivotal in the regulation of granulomatous responses. PMID- 10811281 TI - Use of differential display to detect changes in gene expression in the intermediate snail host Biomphalaria glabrata upon infection with Schistosoma mansoni. AB - Changes in gene expression in Biomphalaria glabrata following infection with Schistosoma mansoni have been investigated using a modified differential display approach. RNA was extracted from ovotestis, mantle tissue and anterior nephridium of control and exposed snails at 2 time-points (4 h and 24 h) post-exposure and analysed by RNA fingerprinting. A number of transcripts were identified; some novel and some homologous to mRNAs in GenEMBL that were previously unknown in B. glabrata. Down regulation of one 241 bp mRNA expressed sequence fragment - with an open reading frame showing 48% identity to a cytochrome p450 over 80 residues has been confirmed using semi-quantitative RT-PCR. Preliminary classification of B. glabrata cyp450 sequence shows it to fall into CLAN 2 of the cytochrome p450 superfamily. Differential display has been successful in identifying changes in gene expression in Biomphalaria glabrata upon infection with Schistosoma mansoni and promises to be a useful technique for the investigation of the interaction between host and parasite. PMID- 10811282 TI - Light-mediated host searching strategies in a fish ectoparasite, Argulus foliaceus L. (crustacea: branchiura). AB - Argulus foliaceus, an obligate fish ectoparasite, can search for its hosts in both light and dark conditions and uses vision in the light. We have examined what searching mode is used at night, when the infection rate was at its highest, and which stimuli produced by the fish are most important. A change of illumination produced a clear difference in the searching behaviour of adult Argulus females. The mean swimming speed and the area explored were 3-4 times higher in the dark, when the parasite employed a cruising search strategy. This changed to an ambush (hover-and-wait) strategy in the light. The swimming activity is accompanied by changes in metabolic costs; the activity of the electron transport system being approximately 25 % lower in the light. The most pronounced light-induced differences in host-searching behaviour took place in moderately hungry parasites (starved for 24-96 h). Less motivated (just having left a fish) or exhausted animals did not exhibit any clear differences in swimming speed. Among the external signals tested, fish smell, from both perch (Perca fluviatilis) and roach (Rutilus rutilus), induced an elevated swimming speed of the parasite. Periodic water movements caused similar but weaker effects. The effects of these stimuli were observed under both light and dark conditions. We conclude that host-searching behaviour of A. foliaceus is under internal (state of hunger) and external (illumination and host-induced signals) control and involves all its sensory equipment (vision, olfaction and mechano reception). Perch (but not roach) reduced their swimming speed in the dark, which make them more susceptible to cruising Argulus. Thus the behavioural interplay between hosts and parasites can also influence the infection rate of A. foliaceus found on perch and roach in Finnish lakes. PMID- 10811283 TI - Functional ryanodine receptor channels in flatworm muscle fibres. AB - Caffeine, which stimulates intracellular Ca2+ release channels known as ryanodine receptor (RyR) channels, induces contraction of individual muscle fibres dissociated from the trematode Schistosoma mansoni, and the turbellarians Dugesia tigrina and Procerodes littoralis. Caffeine is much more potent on S. mansoni fibres (EC50 0.7 mM) than those from D. tigrina or P. littoralis (3.2 mM and 4.6 mM, respectively). These caffeine-induced contractions are blocked by ryanodine, confirming the presence of functional RyR channels in these flatworm muscles. However, the contractions are not blocked by typical RyR channel blockers ruthenium red or neomycin, indicating that there may be important pharmacological differences between the RyR channels in this early-diverging phylum and those of later animals. These studies demonstrate that RyR channels are present in the muscle of these flatworms, and that the sarcoplasmic reticulum stores sufficient Ca2+ to support contraction. PMID- 10811284 TI - Preliminary study of the role of red foxes in Echinococcus multilocularis transmission in the urban area of Sapporo, Japan. AB - In order to assess the infection risk of alveolar echinococcosis among urban residents of Sapporo, the capital of Hokkaido, Japan, a survey was conducted on fox distribution in the urban area and on the prevalence of Echinococcus multilocularis among the foxes. The fox distribution, evaluated from fox footprints left on the snow in parks and woodlands, and from locations of fox carcasses recorded by the Sapporo municipality, was concentrated along the border of the urban area and in the southwestern part of the city, facing the mountain. Fox faeces were collected around active fox dens, and analysed by a coproantigen detection assay and parasite egg examination for the Echinococcus infection. Thirty-three out of 155 faeces were coproantigen positive. Coproantigen-positive faeces were collected from 11 den sites (57.9% of total den sites), and all except 1 were located in the urban fringe. A high intensity of taeniid eggs (> 100 eggs per 0.5 g) containing faeces were also collected in the 3 sites of them. Although Echinococcus infection in rodents was not observed from the necropsy of 23 rodents captured around active fox dens, arvicolid rodents, a suitable intermediate host for E. multilocularis, were captured in the urban fringe. Therefore, the urban fringe offers suitable conditions in which the life-cycle of E. multilocularis could be maintained. Prompt measures to control echinococcus infection should be taken, even in urban areas. PMID- 10811285 TI - The evolutionary ecology of host-specificity: experimental studies with Strongyloides ratti. AB - Factors constraining the evolution of host-specificity were investigated using a gastrointestinal parasitic nematode, Strongyloides ratti. S. ratti is a natural parasite of rats which can also reproduce, with decreased success, in laboratory mice. Observed host-specificity arose from lower establishment, reduced per capita fecundity and more rapid expulsion of parasites from mice relative to rats. Variation in the efficacy of thymus-dependent immunity between host species (rats and mice) was insufficient to explain the majority of the observed differences in parasite establishment and reproductive success. The role of natural selection in determining host-specificity was addressed using experimental selection followed by reciprocal fitness assays in both host species. Experimental selection failed to modify the host-specificity of S. ratti to any measurable degree, suggesting either a lack of genetic variation for this trait or the involvement of as yet unidentified factors underlying the differences in S. ratti fitness in rats and mice respectively. These results are discussed in relation to competing theoretical models of ecological specialization, host immunology and previous attempts to experimentally alter the host-specificity of parasitic nematodes. PMID- 10811286 TI - Potential involvement of tumor suppressor gene expression in the formation of estrogen-inducible pituitary tumors in rats. AB - While pituitary tumors can be induced in rats by the administration of estrogen, susceptibility to such tumors is highly strain dependent. In this study, 21-day old male rats of two strains-Fischer 344 (F344) strain, which is particularly susceptible to pituitary tumors, and Sprague-Dawley (SD) strain, which is relatively resistant, were treated with diethylstilbestrol (DES) over a period of 10 days. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to analyze the expression levels of two tumor suppressor genes, p53 and rb, in the pituitaries. In SD rats, both p53 and rb mRNA appeared to increase in response to DES treatment, while in F344 rats they remained undetectable. Western blot analysis revealed that protein levels of cyclin D, which is a cell cycle regulating protein thought to be a potential oncogene, decreased in response to DES treatment in F344 rats but remained constant in SD rats. The observed differences in the expression levels of p53, rb and cyclin D suggest that they might be involved in the primary process of estrogen-induced pituitary tumor development prior to detectable tumor growth. PMID- 10811287 TI - Effects of third ventricle injection of norepinephrine analogue on LH secretion of the non-laying SIJI goose. AB - The effect of norepinephrine analogue (NA) and norepinephrine antagonists on luteinizing hormone releasing hormone (LHRH) secretion in geese was investigated by monitoring serum levels of LH. Twenty non-laying geese were selected and randomly assigned to 4 groups of 5 each. The geese received a single intraventricular injection of saline (control), NA (12 microg/6 microl), phentolamine (1 microg/6l) plus NA or yohimbine (10 microg/6 microl). Phentolamine, a non-selective adrenoreceptor blocker was given 5 min before NA, but yohimbine, an alpha2-adrenoreceptor blocker, was delivered without NA injection. The results showed that serum levels of LH began to increase 20 min after injection of NA, and that high serum levels of LH were maintained until 105 min, followed by a decline to the basal level 125 min later. Significantly high levels of serum LH were observed at 50 and 105 min after the injection as compared with the respective control. Treatment with phentolamine completely blocked the stimulatory effect of NA on serum levels of LH in NA treated geese. Serum levels of LH increased by yohimbine treatment 5 min after the injection and the high levels of serum LH were maintained until 125 min later. Five out of 8 points were significantly high as compared with their respective control values. The present study also suggested that there is presynaptic modulation in the a noradrenergic neural pathways which mediate the release of LHRH, and that this modulation may be carried out by alpha2-adrenoreceptor blocker. PMID- 10811288 TI - Immunoreactive pit-1 protein in hyperplastic pars intermedia induced by calcitonin of the rat pituitary gland. AB - To elucidate the effects of synthetic salmon calcitonin (sCT) on the cells in the rat pituitary gland, we histopathologically and immunohistochemically examined the early changes after 4 or 13 weeks treatment with sCT 120 IU/kg. Focal proliferative lesions of the anterior pituitary glands were consistently found after treatment with sCT for 13 weeks. Histologically, the cells with the focal proliferative lesions were classified into the following three groups: 1) enlarged basophilic cell focus, 2) vacuolated cell focus and 3) chromophobe cell focus. These focal proliferative lesions had positive staining only for the alpha subunit and failed to show Pit-1 protein immunoreactivity. The sCT treatment also increased the thickness of the pars intermedia. Hypertrophy of the pars intermediate cells was characteristically seen. Furthermore, Pit-1 protein immunoreactivity was clearly detected in the nuclei of the hyperplastic pars intermediate cells. All pars intermediate cells were equally stained by alpha- or beta-MSH and beta-endorphin in both vehicle- and sCT-treatment. No difference was seen. These findings strongly suggest a very close relationship between Pit-1 protein immunoreactivity and cellular proliferation induced by sCT. PMID- 10811289 TI - Thiazolidinediones exert different effects on insulin resistance between dexamethasone-treated rats and wistar fatty rats. AB - We determined the in vivo effects of thiazolidinediones on insulin resistance induced by dexamethasone (Dx), as well as that observed in Wistar fatty (WF) rats, using glucose clamp technique to measure glucose uptake (Gu) and percent suppression of hepatic glucose output (HGOsup) to evaluate insulin resistance. Male Wistar rats were treated with Dx (0.5 mg/kg/day) for 7 days. Pioglitazone (P) or troglitazone (T) was coadministered orally for the same period at 10 and 200 mg/kg/day, respectively. Two, 5 and 20 mU/kg/min. of insulin infusion rates (IIR) were used. The Gu levels at clamp steady-state at IIR20 in rats treated with Dx (16.4 +/- 4.7 mg/kg/min.) were significantly lower than those in control rats (36.3 +/- 2.4). The Gu levels at the same IIR in rats coadminstered with P (19.6 +/- 3.2) and T (21.3 +/- 6.3) were slightly but significantly higher than that in rat treated with Dx. HGOsup at IIR5 in control rats (97.5 +/- 6.2%) was decreased by Dx treatment (52.1+/- 31.3). This decrease was slightly but significantly ameliorated by addition of T (78.3 +/- 12.2). The Gu levels at IIR20 in WF rats (6.6 +/- 0.9) were decreased significantly from that in lean littermates of WF (WL) rats (25.8 +/- 2.1). This attenuation of Gu increase was completely ameliorated with administration of P (20.9+/-2.8) or T (22.2+/-3.9). The HGOsup at IIR20 in WF rats (17.4 +/- 11.2) was significantly decreased from that in WL rats. Administration of P or T ameliorated this decrease completely. These results indicate that Dx induces insulin resistance by mechanisms different from those in WF rat, hence thiazolidinedione administration can be only partially useful to treat insulin resistance induced by Dx. PMID- 10811290 TI - The effect of calcium-sensing receptor gene polymorphisms on serum calcium levels: a familial hypocalciuric hypercalcemia family without mutation in the calcium-sensing receptor gene. AB - Familial hypocalciuric hypercalcemia (FHH) is a benign syndrome with elevated levels of serum calcium, relative hypocalciuria, and non-suppressed serum parathyroid hormone (PTH) levels. FHH usually occurs by a heterozygous mutation of the calcium sensing receptor (Casr), but some FHH patients show no mutations of the Casr. We encountered a unique FHH family in which the proband and her mother had many calcium deposits on their skin. The proband was medicated with Levothyroxine for hypothyroidism due to an iodine transport defect (ITD). We searched for mutation of the Casr, but found none. The only change distinguishing the proband and her mother from her father was at codon 990, reported to be a polymorphic site. We investigated the frequency of polymorphism at codon 990, but a significant relationship between the three genotypes and the serum calcium concentration was excluded. At the other polymorphic sites at codon 536, 926, 986, and 1011, polymorphisms were rare in Japanese, and we could not confirm a significant relationship. In conclusion, mutation in the Casr gene alone does not explain all cases of FHH. The other mechanisms should be identified. PMID- 10811291 TI - Different phenotypes of multiple endocrine neoplasia type 1 (MEN1) in monozygotic twins found in a Japanese MEN1 family with MEN1 gene mutation. AB - We report monozygotic twins who showed different MEN1 phenotypes. The proband (28 y.o., female) had both primary hyperparathyroidism (PHP) and insulinoma, and genetic analysis revealed a point mutation (569del1, exon 3) of the MEN1 gene. This mutation causes a frameshift and produces a stop codon at codon 184. Restriction digestion (HinfI) analysis confirmed the same mutation of the MEN1 gene in six of the affected members including her two sisters, the monozygotic twins, and no such mutation in two unaffected members. In two generations of this family, eight of eleven family members had PHP and four of them were found to have other MEN1-related lesions. Both of the monozygotic twins had PHP. Interestingly, one had pancreatic tumor but the other had no evidence of it. Pituitary MRI showed no pituitary lesion in either of them. This is the first Japanese case of monozygotic twins with different MEN1 phenotypes. PMID- 10811292 TI - Pseudomalabsorption of levothyroxine: a case report. AB - A 51-year-old woman who had been treated with levothyroxine sodium because of hypothyroidism after total thyroidectomy for thyroidal cancer was admitted to our hospital for persistent hypothyroidism despite large dose administration of levothyroxine (600 microg/day). The patient complained of severe general fatigue and body weight gain. Free thyroxine, free triiodothyronine and thyrotropin levels were 0.97 ng/dl, 1.55 pg/ml and 24.51 microU/ml, respectively, under oral administration of levothyroxine. Levothyroxine loading test performed by liquid form, pulverized tablets via nasogastric tube and intravenous administration revealed no evidence of malabsorption or metabolic disorder of levothyroxine, although oral intake of tablets was ineffective due to her factitiousness. We report here a possible case of "pseudomalabsorption of levothyroxine" to emphasize the clinical recognition of this disorder in patients with resistant hypothyroidism. PMID- 10811293 TI - A case of hypothalamic adrenal insufficiency manifested normal ACTH response to insulin-induced hypoglycemia. AB - A low plasma ACTH response to insulin-induced hypoglycemia and an exaggerated and delayed plasma ACTH response to CRH stimulation have been considered as an indicator of hypothalamic hypopituitarism. We report a case of hypothalamic adrenal insufficiency which manifested normal ACTH response to insulin-induced hypoglycemia. This case provides important information to categorize hypothalamic adrenal insufficiency caused by abnormal regulation of CRH release. PMID- 10811294 TI - IgG1 is the dominant subclass of antibody against glutamic acid decarboxylase among type 1 diabetes in Japanese. AB - Autoantibody against glutamic acid decarboxylase (GADA) is a highly sensitive predictor of insulin-dependency in adult diabetic patients as well as young individuals. A considerable number of diabetics who do not reach the insulin dependent stage have this antibody. Recently, type 1 diabetes has been thought to be caused by T helper 1 (Thl)-type autoimmunity based on studies in non-obese diabetic mice, but it is still difficult to investigate antigen-specific T-cell function in human type 1 diabetes. We therefore assessed an IgG subclass assay for GADA, which should reflect T-helper function against GAD. Sera from 14 type 1 diabetic patients positive for GADA by radioligand binding assay were tested for the IgG subclass of GADA. The assay was based on an enzyme-linked immunosorbent assay, which showed a good correlation with radioligand binding assay. The sera of all but one of the 14 type 1 diabetic patients (93%) were positive for the IgG1 subclass of GADA. The IgG2 and IgG3 subclasses of GADA were also detected in one diabetic patient each who were also positive for IgG1. The IgG4 subclass was not detected in any of the sera we tested. We concluded that IgG1 is the dominant subclass of GADA in Japanese type 1 diabetic patients. PMID- 10811295 TI - Involvement of AP-1 and steroidogenic factor (SF)-1 in the cAMP-dependent induction of human adrenocorticotropic hormone receptor (ACTHR) promoter. AB - Adrenocorticotropic hormone receptor (ACTHR) is expressed predominantly in the adrenal glands, and its expression is upregulated by its own ligand, ACTH, via a cAMP-dependent pathway. In the present study, we characterized the 5'-regulatory region of human ACTHR gene to elucidate the molecular mechanisms underlying its adrenal-specific and ACTH/cAMP-dependent expression. The promoter region ( 1017/+47 when the transcription start site is regarded as + 1) and its serial 5' deletions (-764/+47, -503/+-47, -214/+47 and -56/+47) were ligated into the upstream of a luciferase (luc) reporter gene. These constructs were transfected into adrenocortical Y1 cells or non-adrenal JEG3 and Cos-1 cells. In all the cell lines, the luc activity gradually increased with serial 5'-deletions and the maximum activity was conferred by - 56/+ 47. However, the magnitude of luc activity of each deletion construct in non-adrenal cells was much less than that in Y1 cells, suggesting that the promoter functions in an adrenal-specific manner. We identified two Steroidogenic Factor (SF)-1-binding sites at -209 and 35. Electrophoretic mobility shift assay (EMSA) demonstrated that both sites bind to SF-1. Mutation of both sites significantly decreased the activity of -214/+47 promoter in Y1 cells. Transfection of SF-1-expressing plasmid into non-adrenal cells significantly increased the promoter activity, suggesting that SF-1 plays a role in the tissue-specific expression of human ACTHR gene. We identified the region, -764 to -503, that was required for the forskolin/cAMP responsiveness of the promoter. This region contains one AP-1 site. EMSA revealed that the binding of AP-1 to this site increased significantly upon treatment of Y1 cells with forskolin. Mutation of the site abolished the forskolin-responsiveness. In non adrenal cells, the forskolin-responsiveness was observed only when SF-1 expressing plasmid was cotransfected. This is the first demonstration that both AP-1 and SF-1 are required for the cAMP-dependent induction of human ACTHR gene. PMID- 10811296 TI - Reverse correlation between urine nitric oxide metabolites and insulin resistance in patients with type 2 diabetes mellitus. AB - We studied the possible relationship between nitric oxide (NO) production and insulin resistance in patients with type 2 diabetes mellitus. Urine NO metabolites (NOx) were measured as an index for NO production by HPLC combined with a Cd column, Griess reaction and a spectrophotometer in 403 healthy control subjects and 102 hospitalized patients with type 2 diabetes. Glucose infusion rate (GIR) was measured as a reverse index for insulin resistance by euglycemic glucose clamp study using an artificial pancreas in 20 of 102 diabetic patients. Urine NOx was lower in the patients with type 2 diabetes than in healthy control subjects (mean+/-SE: 3.18 +/-0.02 versus 3.25 +/-0.01 log[-micromol/gCr], p<0.01). Urine NOx was correlated with body mass index (BMI) in 102 diabetic patients (r= -0.372, p<0.001), but not related to either age, sex, fasting plasma glucose, HbA1c or blood pressure. Urine NOx was correlated with GIR independent of BMI in 20 diabetic patients (r=0.774, P<0.0001). These findings suggest that NO production is closely related with insulin resistance in patients with type 2 diabetes. PMID- 10811298 TI - DiGeorge syndrome with Graves' disease: A case report. AB - DiGeorge syndrome (DGS) is characterized by aplasia or hypoplasia of the thymus and parathyroid glands, cardiac defects and anomaly face. This syndrome is usually associated with hypocalcemia resulting from hypoparathyroidism. In most cases the initial symptom is tetany caused by hypocalcemia within 24-48 hours after birth, with symptoms by immune abnormality appearing later. We report a woman who passed with no symptoms before age 18 and was diagnosed DiGeorge syndrome by tetany with developing auto-immune thyroid disease (Graves' disease). She had surgery for intraventricular septal defect at age 3, hypoparathyroidism, decrease of T cells in peripheral blood and the deletion of the 22nd chromosome long arm (22q11.2). It is supposed that abnormalities of immune function of this case are not complete as indicated by complicating of Graves' disease, and contributing to her long-term survival. PMID- 10811297 TI - Long-term outcome of endocrine function in patients with neurohypophyseal germinomas. AB - Since neurohypophyseal germinomas occur at the pituitary and hypothalamic axis in children and adolescents, the endocrinopathy is one of the common and critical QOL determinants. We carried out a retrospective study on the outcome of endocrine function in patients with neurohypophyseal germinoma, in order to improve or preserve pituitary function after treatment. Sixteen patients (7 men and 9 women), aged 6 to 26 years were admitted and followed up for 95.3 (14-197) months. DI was noted in 12 patients in pretreatment and 16 in posttreatment regardless of tumor size. We carried out the replacement of GH in all 8 patients, presenting the symptoms under 15 years of age. Gonadal or gonadotropic, thyroid and adrenal hormones were replaced in 9, 12 and 15 patients, respectively. Patients with large tumor compressing chiasm or hypothalamus needed hormonal replacement such as gonadal or gonadotropic and thyroid hormones more frequently (<0.01) than those with small one. In addition, two patients with a small tumor at the pituitary stalk and the 3rd ventricle floor showed the improvement of secretion pattern in gonadotropins and ACTH after chemotherapy, although they later needed radiation therapy to control the tumor. Based on our study and review of literature, the endocrinological studies before and after treatment demonstrated that pituitary dysfunction present before treatment persisted or worsened even after tumor remission, except for patients with small and localized ones. The poor endocrine results is considered to be largely radiation-related. Chemotherapy alone seems to be insufficient to obtain complete response (CR). To avoid radiation related pituitary injury, combination of 24 Gy or less dosage of radiation and appropriate chemotherapy is essential. The earlier diagnosis by repeatedly using neuroimaging and serum and CSF tumor markers and earlier initiation of treatment, before irreversible pituitary-hypothalamic damage occurs, contributes to improvement of the outcome of pituitary functions in patients with neurohypophyseal germinomas. PMID- 10811299 TI - Detection of exogenous growth hormone (GH) administration by monitoring ratio of 20kDa- and 22kDa-GH in serum and urine. AB - We previously demonstrated that individual subjects have fairly constant ratios of serum concentrations of 20 kDa- (20K) and 22 kDa-GH (22K). The aim of this study is to demonstrate the possibility of utilizing the changes in the ratio of 20K/22K for detecting the exogenous administration of 22K. A male patient with idiopathic dilated cardiomyopathy (age 51) received 22K (4U, s.c.) every other day. The concentrations of 20K and 22K in serum and urine were measured using enzyme-linked immunosorbent assays before and after administration. The administration of 22K increased total GH concentration, and markedly decreased the ratio of 20K/22K in serum, especially 2-10 h after the administration. From calculations, it became clear that the concentration of exogenous 22K reached a peak between 2-4 h after the administration and decreased to a negligible level after 24 h. The ratio of 20K/22K in the 0-24 h urine was 5 times lower than that in the 24-48 h urine. These data suggest that, by monitoring the ratio of 20K/22K in serum or urine, it is possible to determine whether or not GH has been externally administered and to calculate the serum GH that has been administered. PMID- 10811300 TI - Transmucosal delivery systems for calcitonin: a review. AB - The commercial availability of peptides and proteins and their advantages as therapeutic agents have been the basis for tremendous efforts in designing delivery systems for such agents. The protection of these agents from biological fluids and physiological interactions is crucial for the treatment efficacy. One such agent is salmon calcitonin, a 32 amino-acid polypeptide hormone used in the treatment of bone diseases such as Paget's disease, hypercalcemia and osteoporosis. Researchers have studied different routes to deliver salmon calcitonin more effectively, including nasal, oral, vaginal and rectal delivery. These systems are designed to protect the polypeptide from the biological barriers that each delivery route imposes. Oil-based and polymer-based delivery systems are discussed. PMID- 10811301 TI - Artificial tear adsorption on soft contact lenses: methods to test surfactant efficacy. AB - Spoilage is a primary factor in the biocompatibility of soft contact lenses (SCL) within the lacrimal fluid. Tears are a complex mixture of proteins, lipids, natural surfactants and salts. The spoilation process is due to a contribution of all these components and of the nature of SCL materials themselves. The aim of this study was to set up methods to observe and quantify lacrimal deposits and to select efficient surfactants for preventing protein deposits. The present study was performed on PMMA-NVP SCL. The behaviour of SCL in presence of tears was studied by means of an in vitro artificial tear model consisting of the main tears components and quantified by a colorimetric technique (BCA) performed directly on the lenses. The nature of the deposit was observed directly by atomic force microscopy (AFM) in a liquid medium showing the same adsorption trend noticed in the quantitative results and identifying specific adsorption sites. The assessment of surfactant adsorption was performed using Maron's method, as a mean to evaluate the affinity of surfactant to the surface, while the action of selected surfactants on pre-treated SCL was assessed using the BCA method. Promising results were obtained with these two different methods which can be used easily for the pre-selection of surfactants for further cleaning solution formulation studies. PMID- 10811302 TI - Marrow stromal osteoblast function on a poly(propylene fumarate)/beta-tricalcium phosphate biodegradable orthopaedic composite. AB - The objective of this study was to assess the osteoconductivity of a poly(propylene fumarate)/beta-tricalcium phosphate (PPF/beta-TCP) composite in vitro. We examined whether primary rat marrow stromal cells would attach, proliferate, and express differentiated osteoblastic function when seeded on PPF/beta-TCP substrates. Attachment studies showed that a confluent monolayer of cells had adhered to the substrates within an 8 h time frame for marrow stromal cells seeded at confluent numbers. Proliferation and differentiated function of the cells were then investigated for a period of 4 weeks for an initial seeding density of 42,000 cells/cm2. Rapid proliferation during the first 24 h as determined by 3H-thymidine incorporation was mirrored by an initial rapid increase in total cell number by DNA assay. A lower proliferation rate and a gradual increase in cell number persisted for the remainder of the study, resulting in a final cell number of 128,000 cells/cm2. Differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity and osteocalcin (OC) production throughout the time course. Both markers of osteoblastic differentiation increased significantly over a 4-week period. By day 28, cells grown on PPF/beta-TCP reached a maximal ALP activity of 11 (+/- 1) x 10(-7) micromol/min/cell, while the OC production reached 40 (+/- 1) x 10(-6) ng/cell. These data show that a PPF/beta-TCP composite exhibits in vitro osteoconductivity similar to or better than that of control tissue culture polystyrene. PMID- 10811303 TI - The kinetics of pentoxifylline release from drug-loaded hydroxyapatite implants. AB - Hydroxyapatite (HAP) was synthesized by the aqueous precipitation method from CaO and H3 PO4 as the reagents. The HAP powders, either subjected or not subjected to preliminary calcination, were mixed with a pore-creating medium and isostatically shaped at a pressure of 350 MPa to form cylindrical samples. A natural product such as flour served as a pore-creating medium. Sintering was performed in the air, at 1200 or 1250 degrees C. The employed procedure allowed for achieving microporous materials of pore sizes ranging from 0.1 to 15 microm and with open porosity values of 23-44%. It was demonstrated that the porosity of the obtained materials depended mainly on the amount of the added pore-creating medium and the temperature of sintering. The implants, shaped as hollow cylinders, were filled with 50 mg of pentoxifylline (PTX) as a model drug. Internal wells for drug placement were drilled in the samples using a high precision drill. The drug release study was performed in pH = 7.35 phosphate buffer, at 37 degrees C. The results showed that the amount and time of PTX release, as well as the lag time were mainly controlled by the open porosity of the carriers. PMID- 10811304 TI - Plasma-sprayed hydroxyapatite (HA) coatings with flame-spheroidized feedstock: microstructure and mechanical properties. AB - Flame-spheroidized feedstock, with excellent known heat transfer and consistent melting capabilities, were used to produce hydroxyapatite (HA) coatings via plasma spraying. The characteristics and inherent mechanical properties of the coatings have been investigated and were found to have direct and impacting relationship with the feedstock characteristics, processing parameters as well as microstructural deformities. Processing parameters such as particle sizes (SHA: 20-45, 45-75 and 75-125 microm) and spray distances (10, 12 and 14 cm) have been systematically varied in the present study. It was found that the increase of particle sizes and spray distances weakened the mechanical properties (microhardness, modulus, fracture toughness and bond strength) and structural stability of the coatings. The presence of inter- and intralamellar thermal microcracks, voids and porosities with limited true contact between lamellae were also found to degrade the mechanical characteristics of the coatings, especially in coatings produced from large-sized HA particles. An effort was made to correlate the effects of microstructural defects with the resultant mechanical properties and structural integrity of the plasma-sprayed hydroxyapatite (HA) coatings. The effects of different heat treatment temperatures (600, 800 and 900 degrees C) on the mechanical properties of the coatings were also studied. It was found that a heat treatment temperature of 800 degrees C does enhance the microhardness and elastic modulus of the coatings significantly (P < 0.05) whereas a further increment in heat treatment temperature to 900 degrees C did not show any discernable improvements (P > 0.1). The elastic response behaviour and fracture toughness of both the as-sprayed and heat-treated HA coatings using Knoop and Vickers indentations at different loadings have been investigated. Results have shown that the mechanical properties of the coatings have improved significantly despite increasing crack density after heat treatment in air. Coatings produced from the spheroidized feedstock of 20-45 microm (SHA 20-45 microm) sprayed at a stand-off distance of 10 cm were found to possess the most favourable mechanical properties. PMID- 10811305 TI - Synthesis and characterization of a novel biodegradable antimicrobial polymer. AB - Bacterial infection is a frequent complication associated with the use of medical devices. In an effort to address this problem, antibacterial agents have been incorporated or applied directly onto the surfaces of numerous types of medical devices. This study assessed the feasibility of using a novel biodegradable polymer to release antibiotic drugs in response to inflammatory related enzymes. A model drug polymer was synthesized using 1,6-hexane diisocyanate (HDI), polycaprolactone diol (PCL), and a fluoroquinolone antibiotic, ciprofloxacin. Polymers were characterized by size-exclusion chromatography (SEC), and elemental analysis. Biodegradation studies were carried out by incubating the polymers with solutions of cholesterol esterase (CE) or phosphate buffer (pH 7.0) for 30 days at 37 degrees C. The degradation was assessed by high-performance liquid chromatography (HPLC), mass spectrometry (MS) and 14C radiolabel release. Subsequently, the activity of the released antibiotic was assessed against a clinical isolate of Pseudomonas aeruginosa. HPLC analysis showed the release of multiple degradation products which were identified, by tandem MS, to include ciprofloxacin and derivatives of ciprofloxacin. The microbiological assessment showed that the released ciprofloxacin possessed antimicrobial activity; 1 microg/ml was measured after 10 days. The results of this study suggest that these novel bioresponsive antimicrobial polymers or similar analogs show promise for use in the control of medical device associated infections. PMID- 10811306 TI - A new peptide-based urethane polymer: synthesis, biodegradation, and potential to support cell growth in vitro. AB - A novel non-toxic biodegradable lysine-di-isocyanate (LDI)-based urethane polymer was developed for use in tissue engineering applications. This matrix was synthesized with highly purified LDI made from the lysine diethylester. The ethyl ester of LDI was polymerized with glycerol to form a prepolymer. LDI-glycerol prepolymer when reacted with water foamed with the liberation of CO2 to provide a pliable spongy urethane polymer. The LDI-glycerol matrix degraded in aqueous solutions at 100, 37, 22, and 4 degrees C at a rate of 27.7, 1.8, 0.8, and 0.1 mM per 10 days, respectively. Its thermal stability in water allowed its sterilization by autoclaving. The degradation of the LDI-glycerol polymer yielded lysine, ethanol, and glycerol as breakdown products. The degradation products of LDI-glycerol polymer did not significantly affect the pH of the solution. The glass transition temperature (Tg) of this polymer was found to be 103.4 degrees C. The physical properties of the polymer network were found to be adequate to support the cell growth in vitro, as evidenced by the fact that rabbit bone marrow stromal cells (BMSC) attached to the polymer matrix and remained viable on its surface. Culture of BMSC on LDI-glycerol matrix for long durations resulted in the formation of multilayered confluent cultures, a characteristic typical of bone cells. Furthermore, cells grown on LDI-glycerol matrix did not differ phenotypically from the cells grown on the tissue culture polystyrene plates as assessed by the cell growth, and expression of mRNA for collagen type I, and transforming growth factor-beta1 (TGF-beta1). The observations suggest that biodegradable peptide-based urethane polymers can be synthesized which may pave their way for possible use in tissue engineering applications. PMID- 10811307 TI - Effect of heat treatment of poly(L-lactide) on the response of osteoblast-like MC3T3-E1 cells. AB - Poly(L-lactide) (PLLA) products are molded by heat extrusion. These treatments may change chemical properties and biological response of the PLLA to cells. In this study, the effect of heat treatment of PLLA on osteoblast proliferation and differentiation was examined in vitro. Osteoblast-like MC3T3-E1 cells were cultured for 2 weeks on the PLLA subjected to various heating temperature and time combinations. The protein, DNA, and hydroxyproline (HYP) contents and alkaline phosphatase (ALP) activity of cells cultured on the untreated (non heated) PLLA with a weight average molecular weight (Mw) of 1,000,000 (high Mw PLLA) were not significantly different from those of cells cultured on glass. The activation of osteoblast differentiation by the high Mw PLLA was weak. In contrast, increases in ALP activity and HYP content were found for cells cultured on the PLLA heated at a high temperature of 200 or 250 degrees C. Heat treatment of high Mw PLLA increased differentiation of MC3T3-E1 cells cultured upon it. Significant degradation of PLLA (decrease in molecular weight and increase in molecular weight distribution) were observed following heat treatment. The Mw of PLLA decreased from 1,000,000 to below 20,000, and 14.4 microg of L-lactic acid was released from 10 mg of PLLA by heating at 250 degrees C. Therefore, the effect of low Mw chemicals, which were expected to be the degradation products of high Mw PLLA after heat treatment, on MC3T3-E1 cell activities was examined. Increases in the protein, DNA and HYP amounts and ALP activity for cells cultured with L-lactide or L-lactic acid were observed at 100 microg/ml, but not at 10 microg/ml. When the cells were cultured on the low Mw PLLA (Mw 20,000), their biological parameters also increased. Twelve micrograms of L-lactic acid released from 10 mg of the low Mw PLLA during 2 weeks incubation. The concentration of L lactic acid in the incubation solution of low Mw PLLA or heat-treated PLLA was too small to cause cell activation. These results suggested that increases in osteoblast differentiation on the heat-treated PLLA was not to due to soluble degradation chemicals, such as L-lactic acid, rather than the remaining low Mw PLLA. PMID- 10811308 TI - Semiautomatic macroencapsulation of large numbers of porcine hepatocytes by coextrusion with a solution of AN69 polymer. AB - We have previously demonstrated that allogenic and xenogenic hepatocytes macroencapsulated manually in AN-69 polymer and transplanted intra-peritoneally in rats remained viable for several weeks. However, this manual technique is inadequate to encapsulate several billions of hepatocytes which would be required to correct hepatic failure in big animals or humans. In the present study, we developed an original semiautomatic device in which isolated pig hepatocytes and the polymer solution containing 6% poly(acrylonitrile-sodium methallylsulfonate), 91% dimethylsulfoxide and 3% 0.9% NaCl solution were coextruded through a double lumen spinneret. The extruded minitube (inner diameter: 1.8 mm, wall thickness: 0.07-0.1 mm) containing the encapsulated hepatocytes fell and coiled up in a 0.9% NaCl solution at 4 degrees C and was cut down in 4 m units containing about 120 million hepatocytes. This process allowed to encapsulate 50 million hepatocytes by minute with a preserved immediate cell viability (92 +/- 5%). To test prolonged cell viability after coextrusion, the minitubes were implanted intraperitoneally in rats. Three and seven days after implantation, they were explanted and analyzed. Cells were viable and well-preserved. Therefore, the semiautomatic device appears able to efficiently macroencapsulate in a limited time several billions of porcine hepatocytes which remain viable after transplantation in xenogenic conditions. PMID- 10811309 TI - Practice of a testing bench to study the effects of cyclic stretching on osteoblast-orthopaedic ceramic interactions. AB - A new experimental method has been used to study the behaviour of human osteoblasts cultured on bioceramics subjected to mechanical strains. The ceramics were alumina, hydroxyapatite (HA) and a duplex system composed of hydroxyapatite covered alumina. The system applied 400 microdeformations for a 6-h period with a cycle frequency of 0.5 Hz to osteoblasts growing on ceramic-covered disks. The effects of strains on short-term cell viability, cell growth, alkaline phosphatase (ALP) activity, and collagen biosynthesis were assessed. When possible, the parameters (lactate dehydrogenase) were studied along the experiment in samples of the culture medium, in the other cases by comparison of stretched and unstretched cultures on the same ceramics with the same cell line. In relationship with the coating, mechanical strains resulted in a decrease in DNA corresponding to cell number, an LDH release during straining, an unchanged (alumina) or decreased (HA and duplex) ALP activity, a decrease (HA and duplex) of collagen and total protein synthesis or an increase of it (alumina). The stress-producing device and its associated protocol are shown to be suitable for investigating the behaviour of cells, cultured on biomaterials subjected to mechanical strain. PMID- 10811310 TI - Tissue responses of calcium phosphate cement: a study in dogs. AB - The in vivo properties of a new kind of calcium phosphate cement were investigated in this study. Calcium phosphate cement was implanted as paste into femoral bone and dorsal muscle of dogs for 3 and 6 months, and as prehardened form into thigh muscles of dogs for 1, 2 and 6 months. Histology was performed on thin un-decalcified sections. No foreign body reaction, no inflammation and no necrosis were found both in bony site and in muscles. There was no connective tissue layer between the cement and bone when cement paste was implanted in the bone. A creeping substitution of cement by bone, in which osteoclast-like cells resorbed the cement as if the cement is a part of bone and new bone was formed directly on the resorption line of calcium phosphate cement, was found. Bone formation was found histomorphologically in pores and deep rugged surface of cement samples (both paste and prehardened form) implanted in muscles of dogs. The induced bone was also identified with backscattered scanning electron microscopy (BSE) and by energy-dispersive X-ray micro-analysis (EDX). The results suggest that the calcium phosphate cement used in this study is biocompatible, resorbable in a manner of creeping substitution, osteoconductive and osteoinductive. It seems that an ideal bone substitute can be developed by using this type of calcium phosphate cement. PMID- 10811311 TI - Osteoconduction at porous hydroxyapatite with various pore configurations. AB - To assess the histological response and the reinforcing effects of bone ingrowth within porous hydroxyapatite (HA) implants depending on pore geometry, four kinds of cylindrical-type with parallel linear pores phi50, 100, 300, 500 microm), one kind of sponge-type with irregular interconnecting pores (phi250 microm) and one cross-type with crossing linear pores (phi100 x 120 microm) of porous HA were prepared. Eighty-four rabbits were divided into six groups, and a 5 x 5 x 7 mm sized porous HA block was inserted through the medial cortical window of the proximal tibia. Histomorphological changes were examined using light and scanning electron microscopy. A biomechanical compression test was performed using material test machines. After implantation, the implants showed different histological changes depending on pore geometry. Active osteoconduction was also found in the phi50 microm sized cylindrical-type porous HA. Evidence of remodeling of new bone and bone marrow formation within porous HA was found in the larger cylindrical-types (phi300, 500 microm), and the sponge- and cross types. The biomechanical test showed that the ultimate compressive strength increased significantly in the phi300 microm sized cylindrical-type, and in the sponge- and cross-types eight weeks after implantation. Porous HA with cylindrical pores could be a useful graft material due to its strength, osteoconductivity and the ease with which its pore geometry can be controlled. PMID- 10811312 TI - Intrahepatic small duct disease of adulthood: a welcome review of a growing topic. PMID- 10811313 TI - Sigmoidoscopy or colonoscopy: which way are we headed? PMID- 10811314 TI - Sleep and functional bowel disorders: can bad bowels cause bad dreams? PMID- 10811315 TI - Retreatment of hepatitis C patients who do not respond to interferon: the search continues. PMID- 10811316 TI - Alcohol: "ice-breaker" yes, "gut barrier-breaker," maybe. PMID- 10811317 TI - Variant forms of cholestatic diseases involving small bile ducts in adults. AB - OBJECTIVE: Cholestasis may result from diverse etiologies. We review chronic cholestatic disorders involving small intrahepatic bile ducts in the adult ambulatory care setting. Specifically, we discuss variant forms of primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) as well as other conditions that may present diagnostic and therapeutic difficulties. METHODS: We conducted a MEDLINE search of the literature (1981-1997) and reviewed the experiences at the Mayo Clinic. All articles were selected that discussed antimitochondrial antibody (AMA)-negative PBC, small-duct PSC (formerly pericholangitis), and idiopathic adulthood ductopenia. RESULTS: The most common chronic cholestatic liver diseases affecting adults are PBC and PSC. Patients without the hallmarks of either syndrome are diagnosed according to their clinical and histological characteristics. Autoimmune cholangitis is diagnosed if clinical and histological features are compatible with PBC but autoantibodies other than AMA are present. Isolated small duct PSC is diagnosed if patients have inflammatory bowel disease, biopsy features compatible with PSC, but a normal cholangiogram. If ductopenia (absence of interlobular bile ducts in small portal tracts) is found histologically in the absence of PSC, inflammatory bowel disease, and other specific cholestatic syndromes such as drug reaction or sarcoidosis, the most likely diagnosis is idiopathic adulthood ductopenia. CONCLUSIONS: Based on these definitions, an algorithm for diagnosis and therapy in patients with laboratory evidence of chronic cholestasis may be constructed, pending results of further investigations into the etiopathogenesis of these syndromes. PMID- 10811318 TI - Medical therapy to reduce postoperative Crohn's disease recurrence. AB - Clinical recurrence of Crohn's disease after surgical resection is a significant problem, with reported rates as high as 55% at 5 yr and 76% at 15 yr. Specific factors that predispose to postoperative recurrence of Crohn's disease have not been well defined. In addition, the underlying pathophysiology of recurrent disease and the reason for its localization to the neoterminal ileum are not well understood. Various operative techniques have been evaluated but none, aside from formation of an ostomy, has been shown to reduce the risk of recurrence. In contrast, there is increasing evidence that postoperative medical therapy has the potential to decrease the risk of postoperative recurrence. Historically, sulfasalazine may have a modest effect on reducing postoperative recurrence of ileal or ileocolonic disease. However, 5-ASA preparations that can selectively deliver mesalamine to the small bowel or anastomotic margin should be more effective. Indeed, in several studies and as confirmed by a meta-analysis, mesalamine has been demonstrated to reduce significantly postoperative recurrence of Crohn's disease. Metronidazole and 6-mercaptopurine or azathioprine also seem to be of benefit in postoperative prophylaxis of disease recurrence, but additional controlled studies are required to define better the efficacy and dose response of these agents. Corticosteroids are ineffective at reducing postoperative recurrence. PMID- 10811319 TI - Approach to colon polyps in the elderly. AB - Colorectal cancer is a common but potentially preventable disease. Nearly all colorectal cancers are thought to arise from adenomatous polyps. The conclusions from the National Polyp Study strongly support the concept that the removal of polyps may prevent the future development of colorectal cancer. With the growing acceptance of screening for colorectal cancer, many elderly patients with adenomatous polyps will be discovered. Because increasing age is a powerful determinant of a higher prevalence of colonic neoplasia in asymptomatic individuals, physicians will need to be prepared to make informed decisions regarding the treatment of elderly patients with colonic polyps. The literature is reviewed and guidelines are formed regarding the optimal surveillance interval for patients with colonic polyps. The age at which surveillance and screening for colorectal neoplasia should stop is also reviewed. Conclusions are based on the currently available data. PMID- 10811320 TI - Safety of a systematic endoscopic biopsy protocol in patients with Barrett's esophagus. AB - OBJECTIVE: Widespread implementation of rigorous, systematic endoscopic biopsy protocols for patients with Barrett's esophagus may be hindered by concerns about their safety. This report describes the safety experience of a large series of patients with gastroesophageal reflux disease and Barrett's esophagus who underwent such procedures. METHODS: Patients in the Seattle Barrett's Esophagus Project undergo biopsy surveillance in a research-based clinical setting, using large channel endoscopes and "jumbo" biopsy forceps. After visual inspection, multiple biopsies are obtained from lesions and at 1- to 2-cm intervals throughout the Barrett's esophageal segment. RESULTS: From 1983 to 1997, 1,458 consecutive endoscopies were performed on 705 patients and 50,833 biopsies (average, 35; maximum, 120 per procedure) were taken. Procedures lasted from 15 to 90 min during which one to two biopsies were obtained per minute. Eleven patients experienced 18 significant adverse events, five of which led to overnight hospitalizations: two for bleeding attributed to concomitant esophageal stricture dilation; two for cardiac dysrhythmias; and one for respiratory arrest. Events managed in outpatient settings included chest pain during seven endoscopies (all accounted for by two patients), chest or epigastric pain developing after five endoscopies, and one tonsillar abrasion. All patients recovered completely, and no deaths, perforations, aspiration, or esophageal stricturing resulted from the procedures. CONCLUSIONS: A rigorous, systematic endoscopic biopsy protocol in patients with Barrett's esophagus does not produce esophageal perforation or bleeding when performed by an experienced team of physicians, nurses, and technicians. PMID- 10811321 TI - Salivary and gastric epidermal growth factor in patients with Zollinger-Ellison syndrome: its protective potential. AB - OBJECTIVE: Evidence is accumulating that epidermal growth factor (EGF) is a major molecule contributing to the maintenance of the integrity of the upper alimentary tract mucosa before and after injury by acid and pepsin. Patients with Zollinger Ellison Syndrome (ZES) typically have hypersecretion of acid and pepsin; however, the concentration and rate of secretion of salivary and gastric EGF that could counteract these potentially aggressive factors are unknown. Accordingly, this study was conducted to determine whether EGF affords mucosal protection in ZES patients. METHODS: The concentration and output of salivary (sEGF) and gastric epidermal growth factor (gEGF) were measured in eight patients with ZES and the results compared to those in 17 patients with nonulcer dyspepsia (NUD), serving as a control group. All patients had normal esophageal and gastric mucosa as determined by endoscopy. Total saliva was collected during 1-h parafilm- and 1-h pentagastrin/parafilm-stimulated conditions, as well as basal and pentagastrin stimulated gastric juice. The concentration and output of EGF were determined by radioimmunoassay. RESULTS: The concentration of EGF in saliva collected from ZES patients after parafilm chewing was significantly higher compared to that in NUD patients (4.61 +/- 0.59 vs 2.75 +/- 0.50 ng/ml, p < 0.05). The concentration of EGF in saliva collected after pentagastrin stimulation in ZES patients was also significantly higher than in NUD patients (4.37 +/- 0.73 vs 2.22 +/- 0.37 ng/ml, p < 0.05). Salivary EGF output during parafilm chewing in ZES and NUD were similar (68 +/- 6.4 vs 109 +/- 25.2 ng/h). Salivary EGF output after administration of pentagastrin in ZES and NUD was also similar (66 +/- 6.1 vs 132 +/- 45.4 ng/h). Basal EGF output in the gastric juice of patients with ZES was 3 fold higher than in patients with NUD (801 +/- 73 vs 271 +/- 32 ng/h, p < 0.01). Pentagastrin-stimulated EGF output was similar in both groups (705 +/- 92 vs 675 +/- 168 ng/h). CONCLUSIONS: Patients with ZES have a significantly higher EGF concentration in saliva and EGF output in basal gastric juice. This elevated content of salivary and gastric EGF in ZES patients may play a protective role in preventing the development of reflux esophagitis and gastric ulcer under the impact of gastric acid and pepsin hypersecretion. PMID- 10811322 TI - Rapid urease tests lack sensitivity in Helicobacter pylori diagnosis when peptic ulcer disease presents with bleeding. AB - OBJECTIVE: The eradication of Helicobacter pylori (H. pylori) in patients with bleeding peptic ulcer disease (PUD) decreases the rate of ulcer rebleeding. Although all methods for H. pylori diagnosis have been extensively evaluated in uncomplicated PUD the efficacy of the commonly used rapid urease test (RUT) has not been established in patients with bleeding peptic ulcer disease. The aim of this study was to evaluate the efficacy of the RUT (CLOtest) in patients with bleeding duodenal ulcers (DUs). METHODS: Consecutive patients with symptoms of upper GI tract hemorrhage and a DU at the time of endoscopy were evaluated. The presence of H. pylori infection was determined by RUT, microbiology, and histology. Consecutive patients with uncomplicated DUs were similarly evaluated. The prevalence of H. pylori as determined by the RUT alone was compared to that determined by a combination of all tests in both patient groups. RESULTS: Fifty five patients with bleeding DUs and 69 with nonbleeding DUs were evaluated. The prevalence of H. pylori in patients presenting with bleeding was 72.7% (95% confidence interval [CI] 61.0-84.5%) and lower than the prevalence rate of 92.8% (95% CI 86.6-98.8%) in patients with uncomplicated PUD (p < 0.05). The prevalence of H. pylori in the bleeding DU group as determined by RUT alone (54.5%) was less than that determined by a combination of all tests (73%) with a false-negative rate of 10 of 40 (25%; 95% CI 11.6-38.4%) (p < 0.05). This false-negative rate was significantly greater than that observed in the group presenting with dyspepsia (1 of 64 [1.6%; 95% CI 0-4.6%]) (p < 0.01). CONCLUSIONS: The prevalence of H. pylori is lower in patients with bleeding DUs when compared to patients with uncomplicated DUs. In addition, the sensitivity and negative predictive value of the RUT is lower in patients presenting with bleeding, and other methods of H. pylori diagnosis should be used in this patient group. PMID- 10811323 TI - Appropriate timing of the 14C-urea breath test to establish eradication of Helicobacter pylori infection. AB - OBJECTIVE: The aim of this study was to determine the performance characteristics of the 14C-urea breath test (UBT) performed 2 wk after the completion of therapy for Helicobacter pylori using a 4 to 6 wk study as the gold standard. METHODS: Patients with active Helicobacter pylori infection at four medical centers received proton pump inhibitor-based triple or quadruple therapy for 10-14 days. Patients underwent the 14C-UBT 2 and 4-6 wk after the completion of therapy. A positive test was defined as 14CO2 excretion of >200 dpm, a negative test as <50 dpm, and an equivocal test as >50 but <200 dpm. Performance characteristics of the 2-wk UBT were calculated using the 4 to 6-wk result as a gold standard. RESULTS: Eighty-five patients were enrolled and 82 patients (mean +/- SD age, 62 +/- 15 yr; 15 women) completed the protocol. Four patients had equivocal UBT results and were excluded from the analysis. Of the 78 patients, 68 (87%) had a negative 4 to 6-wk UBT. The 2-week UBT yielded a sensitivity of 90% (95% confidence interval 72-100%), specificity of 99% (97-100%), and accuracy of 97% (93-100%). In patients with a persistently positive UBT, 14CO2 excretion at 2 wk was significantly lower than at 4-6 wk after therapy (p = 0.03). CONCLUSIONS: A UBT performed 2 wk after therapy yielded results comparable to 4 to 6 wk testing. Further studies to evaluate the optimal time of confirmatory testing in the age of more effective proton pump inhibitor-based triple therapies are warranted. PMID- 10811324 TI - Proximal gastric compliance and perception of distension in type 1 diabetes mellitus: effects of hyperglycemia. AB - OBJECTIVES: Upper GI symptoms and disordered gastric motor function occur frequently in patients with type 1 diabetes mellitus and may be influenced by the blood glucose concentration. The aims of this study were to evaluate proximal gastric compliance and perception of gastric distension during euglycemia and hyperglycemia in unselected patients with type 1 diabetes. METHODS: Ten randomly selected patients with type 1 diabetes were studied. On a single day, isovolumetric and isobaric distensions of the proximal stomach were performed during both euglycemia (blood glucose, 6 mmol/L) and hyperglycemia (15 mmol/L), in randomized order. Sensations of fullness, nausea, and bloating were scored using visual analog scales during each step. Results were compared with those obtained in 10 healthy subjects studied during euglycemia. RESULTS: During euglycemia, perceptions of fullness (p < 0.01), nausea (p < 0.01), and bloating (p < 0.05) were greater during gastric distension in patients with diabetes when compared with healthy controls. In the patients, hyperglycemia increased gastric compliance (p < 0.05) when compared to euglycemia. CONCLUSIONS: In unselected patients with type 1 diabetes 1) the perception of gastric distension during euglycemia is increased compared with healthy controls, and 2) hyperglycemia increases proximal gastric compliance. PMID- 10811325 TI - Defining the role of fiberoptic sigmoidoscopy in the investigation of patients presenting with bright red rectal bleeding. AB - OBJECTIVE: This study was done to determine whether sigmoidoscopy could theoretically constitute sufficient investigation for some patients with bright red rectal bleeding. METHODS: One hundred and forty-three patients undergoing investigative colonoscopy for bright red rectal bleeding and whose source of bleeding was identified were studied. The investigation took place in a large urban hospital over an 11-month period. Data obtained included changes in stool pattern, characteristics of the bleeding, lesions identified, and the distance of the lesion from the anus. RESULTS: In patients younger than 55 yr, all serious lesions except for one malignancy in a patient with massive bleeding lay within 60 cm of the anus and theoretically within reach of the fiberoptic sigmoidoscope. The mixing of red blood with stool was commonly due to distal lesions, especially hemorrhoids. CONCLUSIONS: In young persons with bright red rectal bleeding, fiberoptic sigmoidoscopy may prove to constitute appropriate initial investigation. PMID- 10811326 TI - Clinical utility of endoscopic ultrasound and endscopic ultrasound-guided fine needle aspiration in retroperitoneal neoplasms. AB - OBJECTIVE: Nonpancreatic, retroperitoneal tumors are a relatively uncommon clinical problem. With the advent of endoscopic ultrasound and endoscopic ultrasound-guided fine needle aspiration, gastroenterologists may be called upon to assist in the diagnosis and management of these lesions. This paper reviews the spectrum of upper retroperitoneal lesions encountered at a single institution using endoscopic ultrasound. METHODS: We reviewed all cases of nonpancreaticobiliary or nonadrenal retroperitoneal lesions prospectively gathered from our endoscopic ultrasound database from April 1995 to September 1999. RESULTS: Of 1120 upper endoscopic ultrasound examinations, 18 (1.6%) involved a retroperitoneal lesion; 16/18 lesions were neoplasms, nine were primary retroperitoneal tumors (four lymphomas, two leiomyosarcomas, two extraadrenal paraganglionomas, one leiomyoma), and seven were metastatic cancers. There was one fibrous mass and one mass-like abscess. Endoscopic ultrasound guided fine needle aspiration was used in 15/18 cases. The management of 16 patients was significantly affected by the results of endoscopic ultrasound and biopsy. CONCLUSIONS: Although it is not a frequent indication, assessing upper retroperitoneal tumors with endoscopic ultrasound and endoscopic ultrasound guided fine needle aspiration can significantly affect the subsequent management of patients with these lesions. PMID- 10811327 TI - Sleep disturbances in clinic patients with functional bowel disorders. AB - OBJECTIVE: Sleep deprivation can lower visceral perception thresholds and nonregenerative sleep has been implicated as an etiological factor in chronic hyperalgesia syndromes. The aims of our study were to quantify the self-reported prevalence and type of sleep disturbances in patients with different functional bowel disorders (FBD) and to determine if this prevalence is related to involvement of the upper or lower gastrointestinal (GI) tract, perceived disease severity, or psychological comorbidity. METHODS: We enrolled 505 new FBD patients from an academic referral center specializing in functional GI disorders and 247 community based healthy controls. All patients and controls were prospectively evaluated by validated bowel symptom and sleep questionnaires. A psychological profile was obtained by SCL-90R. RESULTS: We found that 68% of functional dyspepsia (FD), 71.2% of irritable bowel syndrome (IBS)+FD, 50.2% of IBS, and 55.1% of the normal subjects reported having sleep disturbances. Waking up repeatedly during the night and waking up in the morning feeling tired or not rested were the most commonly reported sleep patterns; 57.2% of the patients reported that their abdominal ache awakened them from sleep during the night. Self-reported sleep disturbance was directly related to the perceived intensity of GI symptoms. Self-reported sleep disturbances were equally common in both male (57%) and female (58.4%) FBD patients. There was no significant difference between the mean anxiety and depression scores between patients with and without sleep dysfunction. CONCLUSIONS: FD patients, but not IBS patients, reported sleep disturbances more frequently than healthy control subjects. Abdominal pain or discomfort that awaken FBD patients from sleep during the night were common, and thus a poor discriminating factor between organic and functional disorders. PMID- 10811328 TI - Mycophenolate mofetil versus azathioprine in patients with chronic active ulcerative colitis: a 12-month pilot study. AB - OBJECTIVE: Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) of unknown etiology frequently requiring long-term therapy for control of symptoms and prevention of relapse. Azathioprine (AZA) has been shown to be effective and safe in the treatment of chronic active UC. However, the alternatives to treatment with AZA are limited. Our aim was to compare the efficacy and safety of treatment with mycophenolate mofetil (MMF)/prednisolone versus standard immunosuppressive treatment with azathioprine (AZA)/prednisolone in patients with chronic active UC. METHODS: The study was designed as an open comparison of MMF versus AZA. Twenty-four patients with active UC (Rachmilewitz score > or =6 points) were randomly assigned to the MMF (20 mg/kg)/prednisolone or AZA (2 mg/kg)/prednisolone group. The initial prednisolone dosage was 50 mg and was tapered according to a standard protocol. Treatment was scheduled for 1 yr. RESULTS: The rates of remission were higher in the AZA/prednisolone group (n = 12) than in the MMF/prednisolone group (n = 12) throughout the study. Remission rates were 92% versus 67% after 4 wk, 92% versus 67% after 3 months, 92% versus 67% after 6 months, 83% versus 78% after 9 months, and 100% versus 88% after 1 yr. The number of patients not requiring steroids was higher in the AZA/prednisolone group than in the MMF/prednisolone group. Moreover, in the AZA/prednisolone group no severe adverse events were recorded, whereas in the MMF/prednisolone group two severe adverse events were observed: one patient discontinued MMF after 6 months because of recurrent upper airway infections, and one patient exhibited a bacterial meningitis after 9 months. CONCLUSIONS: Treatment with AZA/prednisolone appears to be more effective and safe compared to MMF/prednisolone in patients with chronic active UC. MMF might be an alternative treatment for patients with contraindications to AZA. To further evaluate the effects of MMF in active UC, a placebo-controlled double-blinded study appears warranted. PMID- 10811329 TI - Are 99mTc leukocyte scintigraphy and SBFT studies useful in children suspected of having inflammatory bowel disease? AB - OBJECTIVE: The goal of this retrospective study was to assess whether 99mTc-white blood cell (WBC) scintigraphy and upper gastrointestinal small bowel follow through (UGI-SBFT) could exclude inflammation in children suspected of having inflammatory bowel disease (IBD). METHODS: Of a population of 313 children who had a 99mTc-WBC scan, 130 children were studied exclusively to rule out IBD. Sixty-nine colonoscopies with biopsies were done within a short time interval of the 99mTc-WBC scans. There were also 51 controls studied with 99mTc-WBC scintigraphy. RESULTS: Of the 130 children studied to exclude IBD, the final diagnosis was Crohn's disease in 27, ulcerative colitis in nine, miscellaneous colitis in 13, probably normal in 42, and normal in 39. The 99mTc-WBC scans were positive in all but three newly diagnosed Crohn's disease, ulcerative colitis, or miscellaneous colitis children. The false-negative 99mTc-WBC studies were seen in children with mild inflammation on biopsies and normal UGI-SBFT studies. In the 46 children with a true-positive 99mTc-WBC scan, 81% (17/21) of UGI-SBFT studies were normal. In five children with equivocal UGI-SBFT studies, the 99mTc-WBC scan correctly predicted if inflammation was present in the terminal ileum. CONCLUSIONS: Our results suggest that 99mTc-WBC is useful as an initial screening modality to exclude IBD, and is more sensitive than UGI-SBFT studies. PMID- 10811330 TI - Stress and exacerbation in ulcerative colitis: a prospective study of patients enrolled in remission. AB - OBJECTIVE: We sought to determine whether psychosocial factors influence the course of ulcerative colitis, hypothesizing that high perceived stress among patients with inactive disease will increase the risk of subsequent exacerbation. METHODS: Sixty-two patients with known ulcerative colitis were enrolled into a prospective cohort study while in clinical remission. Their perceived stress, depressive symptoms, and stressful life events were followed, along with potential confounders, for up to 45 months; exacerbation status was monitored for up to 68 months. RESULTS: The 27 patients who experienced an exacerbation were compared with those who remained in remission. Having a score in the upper tertile on the long-term (past 2 yr) baseline Perceived Stress Questionnaire significantly increased the actuarial risk of exacerbation (hazards ratio = 2.8, 95% confidence interval 1.1-7.2). At any given study visit, high long-term stress tripled the risk of exacerbation during the next 8 months (risk for the three tertiles, 8.3%, 16.7%, and 26.2%, p = 0.02). Shorter sleep time, briefer remission, histological activity, and use of nonsteroidal antiinflammatory drugs, antibiotics, or oral contraceptives also increased the medium- and/or long-term risk of exacerbation, but adjustment for these variables did not eliminate the associations with stress. Exacerbation was not associated with stressful life events, depressive symptoms, short-term (past month) perceived stress, smoking, disease extent or duration, or severity of recent course. CONCLUSIONS: Short-term stress does not trigger exacerbation in ulcerative colitis, but long-term perceived stress increases the risk of exacerbation over a period of months to years. PMID- 10811331 TI - Mucosal cytokine production in radiation-induced proctosigmoiditis compared with inflammatory bowel disease. AB - OBJECTIVE: We measured the mucosal levels of interleukin 1beta (IL-1beta), IL-2, IL-6, and IL-8 in affected segments of radiation-induced proctosigmoiditis and compared these with the levels in normal controls and inflammatory bowel disease (IBD) patients. METHODS: Thirteen patients with histologically proven radiation proctosigmoiditis, 32 patients with active ulcerative colitis (UC), 35 patients with Crohn's disease, and 15 normal subjects undergoing routine colonoscopy were included in the study. All patients underwent colonoscopy and mucosal biopsies were obtained from both diseased and normal-appearing areas. Mucosal levels of IL 1beta, IL-2, IL-6, and IL-8 were determined by solid-phase enzyme-linked immunosorbent assay (ELISA) using the Quantikine method (R&D Systems, Minneapolis, MN). All the data were statistically analyzed using Student's t test. RESULTS: The mucosal levels of IL-2, IL-6, and IL-8 were significantly higher in both diseased segments (5.62 +/- 0.13, 1.60 +/- 0.31, and 21.45 +/- 4.03 pg/ml, respectively) and normal-appearing segments (3.83 +/- 0.78, 1.36 +/- 0.34, and 13.45 +/- 3.18 pg/mg) in the radiation proctitis group compared to those of normal control subjects (1.74 +/- 0.23, 0.67 +/- 0.09, and 4.99 +/- 1.39 pg/mg). These differences were statistically significant (p < 0.05). In the UC group, IL-1beta, IL-6, and IL-8 levels in diseased segments were 4.98 +/- 0.53, 2.22 +/- 0.28, and 88.85 +/- 8.05 pg/mg, respectively. In Crohn's disease patients, IL-1beta, IL-6, and IL-8 levels were 5.45 +/- 0.93, 2.88 +/- 0.58, and 61.68 +/- 10.02 pg/mg, respectively. All these levels were significantly higher (p < 0.05) compared with IL-1beta, IL-6, and IL-8 levels from normal segments of IBD patients. Compared with the radiation proctitis patients, the levels of IL-6 and IL-8 were significantly higher in the IBD group. CONCLUSIONS: The mucosal levels of IL-2, IL-6, and IL-8 were significantly higher in both diseased and normal segments of colon in patients with radiation proctitis, compared with normal controls. Only IL-1beta levels were significantly higher in diseased segments, compared with endoscopically normal-appearing segments in radiation proctitis. These results indicate that there is a similarity in the activation of mucosal cytokines between IBD and radiation proctosigmoiditis. This may partly explain the beneficial effects of similar topical and systemic agents such as steroids and mesalamine compounds when used in radiation-induced proctosigmoiditis. PMID- 10811332 TI - Persistence of abnormal gastrointestinal motility after operation for Hirschsprung's disease. AB - OBJECTIVE: Recent studies in patients with Hirschsprung's disease (HD) suggest that morphological abnormalities of the intramural intestinal plexuses are not restricted to the colon. In this report, symptoms and objective tests of gastrointestinal (GI) motor dysfunction were determined long after operative treatment to see whether evidence of a more widespread and relevant motility disturbance could be detected. METHODS: Twenty-one children were available for study an average of 6.6 yr after surgery for HD. All of these patients underwent evaluation of bowel frequency per week, total GI transit time (TGTT), and a scintigraphic gastric emptying test using solid food; anorectal manometry and segmental colonic transit times were performed in a subset of patients. Results were compared with findings in appropriately matched controls. RESULTS: Frequency of defecation per week in patients with HD after surgery was not different from that in control children, but TGTT was significantly longer (p < 0.01). Percentage retention of gastric isotope at 60 min exceeded the normal range in 12 of 21 (57.1%) patients, and colonic transit was abnormal in all six children studied. Symptoms persisted in two-thirds of patients postoperatively, and transit abnormalities were more common in the symptomatic subset (p = 0.026). CONCLUSIONS: Our data show that, in a subset of patients with HD, GI motor dysfunction persists long after surgical correction. The heterogeny of basic defects responsible for HD could provide the substrate for these motor abnormalities that, in turn, seem at least partially responsible for continuation of the symptomatic state. PMID- 10811333 TI - Alteration of intestinal microflora is associated with reduction in abdominal bloating and pain in patients with irritable bowel syndrome. AB - OBJECTIVE: The influence of the gastrointestinal (GI) microflora in patients with irritable bowel syndrome (IBS) has not been clearly elucidated. This study was undertaken to see if patients with IBS have an imbalance in their normal colonic flora, as some bacterial taxa are more prone to gas production than others. We also wanted to study whether the flora could be altered by exogenous supplementation. In a previous study we have characterized the mucosa-associated lactobacilli in healthy individuals and found some strains with good colonizing ability. Upon colonization, they seemed to reduce gas formation. METHODS: The study comprised 60 patients with IBS and a normal colonoscopy or barium enema. Patients fulfilling the Rome criteria, without a history of malabsorption, and with normal blood tests underwent a sigmoidoscopy with biopsy. They were randomized into two groups, one receiving 400 ml per day of a rose-hip drink containing 5 x 10(7) cfu/ml of Lactobacillus plantarum (DSM 9843) and 0.009 g/ml oat flour, and the other group receiving a plain rose-hip drink, comparable in color, texture, and taste. The administration lasted for 4 wk. The patients recorded their own GI function, starting 2 wk before the study and continuing throughout the study period. Twelve months after the end of the study all patients were asked to complete the same questionnaire regarding their symptomatology as at the start of the study. RESULTS: All patients tolerated the products well. The patients receiving Lb. plantarum had these bacteria on rectal biopsies. There were no major changes of Enterobacteriaceae in either group, before or after the study, but the Enterococci increased in the placebo group and remained unchanged in the test group. Flatulence was rapidly and significantly reduced in the test group compared with the placebo group (number of days with abundant gas production, test group 6.5 before, 3.1 after vs 7.4 before and 5.6 after for the placebo group). Abdominal pain was reduced in both groups. At the 12-month follow-up, patients in the test group maintained a better overall GI function than control patients. There was no difference between the groups regarding bloating. Fifty-nine percent of the test group patients had a continuous intake of fermented products, whereas the corresponding figure for the control patients was 73%. CONCLUSIONS: The results of the study indicate that the administration of Lb. plantarum with known probiotic properties decreased pain and flatulence in patients with IBS. The fiber content of the test solution was minimal and it is unlikely that the fiber content could have had any effect. This type of probiotic therapy warrants further studies in IBS patients. PMID- 10811334 TI - Ileocolonic anastomotic ulcers: a case series and review of the literature. AB - OBJECTIVE: Although gastric anastomotic ulcers have been well described, there are only a few reports in the literature, mostly in the pediatric population, of ulcers occurring at colonic anastomotic sites. Our aim was to determine the clinical profile of postoperative colonic anastomotic ulcers in adult patients undergoing colonoscopy at our institution. METHODS: We performed a retrospective review of colonoscopies done at our institution between 1993 and 1997. RESULTS: Six patients with colonic anastomotic ulcers were identified; all had ileocolonic anastomoses. All patients presented with iron deficiency anemia with evidence of gastrointestinal (GI) blood loss and this was the indication for colonoscopy. The age at the time of ileocolonic anastomoses ranged from birth to 74 yr and age at the time of colonoscopy ranged from 24 to 76 yr. The interval between surgery and detection of anastomotic ulcer ranged from 15 months to 28 yr. Only two patients had a possible etiology: one with previous small bowel Crohn's disease, and the other with significant nonsteroidal antiinflammatory drugs (NSAID) use and evidence of small-bowel ulcers in the adjacent ileum as well. Three of the patients had previously undergone surgical resection and revision of the anastomosis without benefit; the ulcers recurred at the new anastomosis and continued to bleed. CONCLUSIONS: Ulcers can develop at sites of ileocolonic anastomoses. The commonest presentation is with iron deficiency anemia due to occult blood loss. The etiology of the ulcer, in most patients, remains speculative. The therapeutic approach to these ulcers is to discontinue all NSAIDs, treat underlying inflammatory bowel disease, if present, and supplement with oral iron. Surgery should be reserved for life-threatening bleeds or for anemia refractory to oral iron therapy. PMID- 10811335 TI - Effects of wheat bran and Olestra on objective measures of stool and subjective reports of GI symptoms. AB - OBJECTIVE: The aim of this study was to compare the effects of two nondigested, nonabsorbed dietary components on objective and subjective measures of gastrointestinal (GI) function. METHODS: A placebo-controlled parallel study compared the effects of wheat bran (20 g/day or 40 g/day in cereal), a well-known dietary fiber, with those of olestra (20 g or 40 g/day in potato chips), a nonabsorbed fat, on stool output, stool apparent viscosity (log peak force for extrusion [PF]), stool water content, and GI symptoms. Sixty subjects resided on a metabolic ward for 9 days: 3 days baseline and 6 days treatment. RESULTS: Compared with placebo, consumption of 20 g/day wheat bran for 6 days resulted in a rapid (within 38 h) increase in mean (+/-SE) stool output (placebo, 150 +/- 29 g/day; bran, 246 +/- 35 g/day, p < 0.05), a directional increase in mean stool water content (placebo, 81.2 +/- 0.8%; bran, 83.9 +/- 0.8%), stool water output (placebo, 159 +/- 54 g/day; bran, 238 +/- 30 g/day), and bowel movement frequency (BM/day) (placebo, 2.2 +/- 0.4; bran, 2.6 +/- 0.4), and no stool-softening effect (placebo log PF, 2.9 +/- 0.1 g; bran log PF, 2.9 +/- 0.1 g). Wheat bran 40 g/day results were not significantly different from wheat bran 20 g/day. Compared with placebo, consumption of olestra 20 g/day and 40 g/day for 6 days showed no significant difference in mean stool output (151 +/- 18 g/day and 204 +/- 28 g/day, respectively), mean BM frequency (1.8 +/- 0.2 BM/day and 2.1 +/- 0.3 BM/day, respectively), and stool water output (138 +/- 13 g/day and 184 +/- 31 g/day, respectively), a significant (p < 0.05) decrease in stool water content (75.5 +/- 1.7% and 72.6 +/- 2.2%, respectively), and either no effect on stool apparent viscosity (olestra 20 g/day, mean log PF, 3.0 +/- 0.1 g) or a gradual stool-softening effect beginning study day 6 (olestra 40 g/day, log PF, 2.7 +/- 0.1 g). None of the treatment groups showed a significant increase in GI symptoms compared with placebo. CONCLUSIONS: Consumption of wheat bran in excess of levels in a typical Western diet significantly increased stool output, but did not soften normal-viscosity stool nor result in an increase in common GI symptoms. The observed plateau effect for wheat bran at 40 g/day suggests a maximal mechanical stimulatory effect. Consumption of olestra in excess of usual snacking conditions did not result in a significant increase in stool output or common GI symptoms. At the highest level tested, olestra resulted in a gradual stool softening effect after several days of consumption. PMID- 10811336 TI - Human recombinant tissue transglutaminase ELISA: an innovative diagnostic assay for celiac disease. AB - OBJECTIVE: Tissue transglutaminase is the autoantigen recognized by the sera of celiac patients. An enzyme-linked immunosorbent assay (ELISA) based on guinea-pig tissue transglutaminase was recently used to measure serum tissue transglutaminase antibodies for the diagnosis of celiac disease. We determine the sensitivity and specificity of an ELISA test based on the use of human recombinant transglutaminase, compared with the guinea pig transglutaminase ELISA and IgA antiendomysium antibodies. METHODS: Serum samples were tested from 65 patients with intestinal biopsy proven celiac disease, from 10 patients with Crohn's disease, and from 150 healthy blood donors. RESULTS: Human transglutaminase ELISA identified 64 of 65 celiac patients, whereas the guinea pig transglutaminase ELISA and IgA antiendomysium antibodies identified 58 of 65 and 60 of 65 subjects, respectively. The three tests showed comparable specificity. CONCLUSIONS: These results proved that the human tissue transglutaminase-based ELISA represents a cost-effective strategy for identifying both symptomatic and atypical forms of celiac disease and could mean that intestinal biopsy need no longer be the gold standard for diagnosing this clinical condition. Furthermore, early identification and treatment of patients with celiac disease in an outpatient setting could have significant implications for reducing long-term morbidity and can produce major savings in future health care costs. PMID- 10811337 TI - Multiple organ dysfunction in ulcerative colitis. AB - OBJECTIVE: The mortality rate in severe ulcerative colitis (UC) is commonly attributed to major colonic complications or surgical procedures. Early recognition of the severity of the colitis, intensive medical treatment, and prompt surgery have all contributed to improving its outcome over the past 40 yr. Recently, we have observed some fatal cases of severe UC in which death was related to multiple organ dysfunction syndrome (MODS). This complication, associated with a very high mortality rate, may occur in several acute critical diseases, both infectious and noninfectious, but has so far not been reported in UC. The aim of this study was to evaluate the prevalence and outcome of MODS in severe UC. METHODS: The records of 180 consecutive patients admitted to the Gastrointestinal Unit, University of Rome for an acute severe attack of UC during the period 1976-1998 were retrospectively analyzed. Severity of UC was defined according to the criteria of Truelove and Witts. MODS was defined according to the original criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference 1992. All patients were on a standard intensive regimen consisting of total parenteral nutrition and hydrocortisone 100 mg q.i.d. Colectomy was performed according to the timing of the Oxford intensive regimen. RESULTS: Of these 180 severe UC patients, 11 (6.1%) experienced clinical and laboratory features of MODS. The lung was involved in five patients, the kidney in three, the liver in seven, the central nervous system in three, the hematological system in three, and the pancreas in one. MODS was preceded by toxic megacolon in five patients and by so-called "impending megacolon" in four, whereas in two patients no previous complications of UC were observed. MODS developed during the first attack of colitis in seven patients and during relapse in four. The overall mortality rate was 12/180 (6.6%). Of the 12 patients who died, eight (72.7%) had MODS. CONCLUSIONS: These data indicate that UC must be included among the causes of MODS. In our referral center for inflammatory bowel diseases, MODS was responsible for the majority of UC cases with a fatal outcome. The timely identification of signs of MODS should prompt admission to an intensive care unit and emergency surgery. PMID- 10811338 TI - A meta-analysis and overview of the literature on treatment options for left sided ulcerative colitis and ulcerative proctitis. AB - OBJECTIVES: Therapeutic trials in left-sided ulcerative colitis (L-UC) and ulcerative proctitis (UP) have lacked control for medication type, dose, delivery, and duration of therapy. METHODS: All published therapeutic articles and abstracts in L-UC or UP from 1958-1997 were reviewed. Improvement, remission rates, and adverse events were recorded for all (ALL), placebo-controlled (PC) studies, and for PC studies passing quality assessment (QA) scoring. Meta analysis was used where appropriate. RESULTS: Left-sided UC: For active disease, 67 studies (17 PC; 10 QA) were identified. Mesalamine enemas achieved remission in a duration but not a dose response (QA), with higher remission rates than steroid enemas (ALL) and clinical improvement rates superior to oral therapies (QA, ALL). Remission maintenance: 17 (six PC, six QA) studies were identified. Mesalamine therapies had comparable remission rates at 6 months, with a possible dose but not delivery effect. Mesalamine enema dosing intervals between QHS to Q3 days maintained efficacy. Reported adverse events were most common with oral sulfasalazine and dose-independent for mesalamine. Withdrawals from therapy were less than placebo, or < or =3%. Ulcerative proctitis: For active disease, 18 (nine PC, three QA) studies were identified. Mesalamine suppositories achieved clinical improvement and remission in a duration but not dose response, with higher rates of remission than topical steroids (ALL). Remission maintenance: three (three PC, two QA) studies were identified. Remission ranged from 75% to 90% (6 months) and 61-90% (12 months) for mesalamine agents. Reported adverse events were most common for mesalamine foam (8%). Withdrawals from therapy were <2%. CONCLUSIONS: In L-UC and UP, the efficacy and side-effect profile of topical mesalamine are dose independent and superior to oral therapies and topical steroids. Economic analysis suggests that use of these agents will also result in an overall decrease in patient costs. PMID- 10811339 TI - Gender distribution in asymptomatic and invasive amebiasis. AB - OBJECTIVE: The majority of individuals infected by the protozoan parasite Entamoeba histolytica experience subclinical infections. However, a small proportion of parasitized individuals develop severe invasive disease such as amebic dysentery or amebic liver abscess. Invasive amebiasis affects predominantly men; the usual explanation for this has been that men have a higher rate of asymptomatic infections and therefore experience a higher rate of invasive disease. To date, there is no convincing evidence of an increased rate of asymptomatic infection of men as compared with women. The purpose of this study was to evaluate the evidence supporting the hypothesis that men have higher rates of asymptomatic infection and thus an increased frequency of invasive amebiasis. METHODS: We reviewed published reports of invasive amebiasis and population-based parasitological studies from 1929-1997 to compare the gender ratio of asymptomatic and symptomatic E. histolytica infection. Infections with E. histolytica were differentiated from the nonpathogenic E. dispar whenever possible. RESULTS: The reports of invasive amebiasis (dysentery, liver abscess, colonic perforation, peritonitis, appendicitis, and ameboma) showed a higher proportion of men than women (ratio, male:female = 3.2:1, p < 0.05). This contrasts with the epidemiological surveys, where the rate of asymptomatic infection with E. histolytica was the same (1:1) for both genders (p > 0.05). CONCLUSIONS: Asymptomatic E. histolytica infection is equally distributed between the genders. The high proportion of men with invasive amebiasis may be due to a male-related susceptibility to invasive disease. PMID- 10811340 TI - Interferon and ribavirin versus interferon and amantadine in interferon nonresponders with chronic hepatitis C. AB - OBJECTIVE: The aim of this study was to compare the potential efficacy and safety of a combination of interferon and ribavirin with that of interferon and amantadine in patients who had previously failed to respond to interferon monotherapy. METHODS: A total of 29 patients were randomized to 3 million units of alpha-interferon three times weekly with 1000 mg of ribavirin daily (group A, n = 14) or an identical dose of alpha-interferon and amantadine hydrochloride given in a dose of 200 mg daily (group B, n = 15). Patients were treated for 24 wk and observed for 24 wk posttreatment. RESULTS: The treatment groups were evenly matched with respect to gender, frequency of genotype 1, presence of fibrosis, as well as baseline alanine aminotransferase (ALT) and HCV RNA levels. At the end of therapy, five of 14 or 36% of patients in group A versus 0 of 15 in group B had both normal serum ALT and nondetectable HCV RNA by polymerase chain reaction (p = 0.017). A complete response was sustained, however, in only two of 13 patients (15%) in group A who completed 24 wk of observation posttreatment. CONCLUSIONS: A substantial proportion of interferon nonresponders have an end-of treatment biochemical and virological response to a combination of interferon and ribavirin, and sustained responses are possible. The addition of amantadine to interferon, in contrast, does not seem to enhance the antiviral effectiveness of interferon in patients who have previously failed to respond. PMID- 10811341 TI - Bacterial infection in cirrhotic patients and its relationship with alcohol. AB - OBJECTIVE: Infections are regarded as a major complication and an important cause of death in cirrhotics. Alcohol is a predisposing factor to infections in such patients. This study was undertaken to compare the frequency and evolution of bacterial infection among alcoholic and nonalcoholic cirrhotics. METHODS: To observe this relationship, we retrospectively studied a cohort of 382 cirrhotic inpatients, 201 of whom were alcoholic (alcohol intake > or =80 g/day for > or =10 yr) and 181 of whom were nonalcoholic. RESULTS: A total of 128 (33.5%) patients presented with infection upon hospitalization, 78 of whom were alcoholic and 50 of whom were nonalcoholic (p = 0.02). A total of 157 cases of infection were diagnosed, with spontaneous bacterial peritonitis as the most prevalent one (54.1%), followed by pneumonia (18.5%), infection of the soft parts (10.8%), and urinary tract infection (7.0%). Infection and deaths were more frequent in patients with Child-Pugh C than in those with Child-Pugh A/B (p = 0.003, p = 0.0002 respectively). Alcoholic patients with Child-Pugh A/B were more susceptible to infection compared to nonalcoholic patients (p = 0.02), although no difference was noted as to the number of deaths (p = 0.1). With regard to patients with Child-Pugh C, no statistical difference was found in the infections or deaths among alcoholics and nonalcoholics (p = 0.8, p = 0.8). CONCLUSIONS: Our findings suggest that, despite the fact that bacterial infections are more common in cirrhotic alcoholics, its seems that the mortality rate is associated more with the severity than with the etiology of the hepatic disease. PMID- 10811342 TI - The influence of perioperative blood transfusion on intrahepatic recurrence after curative resection of hepatocellular carcinoma. AB - OBJECTIVE: This study retrospectively evaluated the association between perioperative blood transfusion and intrahepatic recurrence in patients with hepatocellular carcinoma (HCC) who had undergone curative hepatic resections. METHODS: Hepatic resection was performed with curative intent in 195 patients with primary HCC between 1985 and 1996. Patients who had received perioperative blood transfusion (transfused group: n = 117) and those who had no perioperative blood transfusion (nontransfused group: n = 78) were compared in terms of conventional prognostic variables and cancer-free survival by the univariate and multivariate analyses. RESULTS: The 1-, 3-, and 5-yr cancer-free survival rates in the nontransfused and transfused groups were 83.4% and 67.9%, 43.0% and 36.7%, and 23.1% and 24.6%, respectively (p = 0.175). Multivariate analysis of prognostic factors in all patients revealed that vascular invasion, tumor size (> or =5 cm), and Child's class were independent factors for intrahepatic recurrence. Further analyses in various stratified groups showed that perioperative blood transfusion was an independent predictor of prognosis in HCC patients with portal vein invasion (RR: 2.8, p = 0.0038). The 1-, 3-, and 5-yr survival rates in the nontransfused and transfused groups with portal vein invasion were 71.9% and 41.6%, 54.5% and 10.9%, and 26% and 0%, respectively (p = 0.0003). CONCLUSIONS: We conclude that perioperative blood transfusions enhance the risk of intrahepatic recurrence of HCC in patients with portal vein invasion. As well, the more difficult surgery and the increased manipulation of the liver that occur in these cases create a greater possibility of tumor dissemination. PMID- 10811343 TI - Evidence of impaired gallbladder function in patients with liver cirrhosis by quantitative radionuclide cholescintigraphy. AB - OBJECTIVE: The aim of this study was to understand gallbladder function in liver cirrhosis. METHODS: Gallbladder function was measured in 45 patients with liver cirrhosis by Tc-99m-labeled imino-diacetic acid analog cholescintigraphy. Gallbladder function was represented by the filling fraction and the ejection fraction (EF). The patients were divided into three groups, based on cirrhotic severity, using the modified Child's classification: A = good, B = fair, and C = poor. Fifteen normal control subjects were selected for comparison. RESULTS: There were no significant differences in filling fraction among all of the study groups. Normal controls had significantly higher 60-min EF values than did liver cirrhosis patients. Among the liver cirrhosis patients, class A patients had the highest EF values, and class C patients had the lowest EF values. CONCLUSIONS: We found that liver cirrhosis may significantly impair the gallbladder emptying, based on the evidence of quantitative Tc-99m-labeled imino-diacetic acid analog cholescintigraphy findings. PMID- 10811344 TI - Genetic polymorphisms of interleukin-1beta in association with the development of alcoholic liver disease in Japanese patients. AB - OBJECTIVE: Cytokine interleukin-1beta plays a central role in the inflammation process. Serum levels of IL-1beta are elevated in patients with alcoholic liver disease (ALD), especially in those with cirrhosis and alcoholic hepatitis. Recently, the presence of genetic polymorphisms of this cytokine was confirmed. The aim of this study was to determine whether IL-1beta polymorphisms are associated with the development of ALD. METHODS: We examined the frequency of two polymorphisms in the IL-1beta gene located in promoter -511 and exon 5 +3953 locus by restriction fragment length polymorphisms in 142 male patients with ALD, 30 heavy drinkers without ALD, and 218 healthy controls. RESULTS: The carriers of -511 IL-1beta allele 2 were present significantly more often in patients with alcoholic cirrhosis than in those with noncirrhotic ALD (p = 0.026), heavy drinkers without ALD (p = 0.001), and healthy controls (p = 0.032). The frequencies of allele 2 and heterozygotes of +3953 polymorphism were both significantly higher in heavy drinkers without ALD than in patients with ALD (allele, p = 0.030; genotype, p = 0.027) and healthy controls (allele, p = 0.047; genotype, p = 0.043). The haplotype, IL-1beta -511 allele 2/+3953 allele 1 was associated with the development of alcoholic cirrhosis (p < 0.05). CONCLUSIONS: These results suggest that IL-1beta polymorphisms may be related to the development of ALD in Japanese alcoholics. PMID- 10811345 TI - Tattoo application is not associated with an increased risk for chronic viral hepatitis. AB - OBJECTIVE: Many reports cite tattoo application as an independent risk factor for viral hepatitis. The purpose of this study was to determine whether patients with tattoos are at increased risk for chronic viral hepatitis. METHODS: A total of 212 patients, aged 18-55 yr, who presented to the emergency center and outpatient clinic at a suburban tertiary care hospital participated in the study. Of these, 106 had tattoos and 106 did not. No patient had known liver disease or viral hepatitis. Hepatitis B DNA, hepatitis C RNA, and hepatitis G RNA were measured in the serum using the polymerase chain reaction on stored serum samples. Each participant completed an anonymous questionnaire concerning risk factors for viral hepatitis. RESULTS: Patients with tattoos did not have a higher rate of chronic hepatitis B, C, or G than did a gender-matched group without tattoos. One (0.9%), seven (6.6%), and three patients (2.8%) in the tattoo group were positive for hepatitis B DNA, hepatitis C RNA, and hepatitis G RNA, respectively. Among controls, no patients (0%), three (2.8%), and six (5.6%) patients were positive for hepatitis B DNA, hepatitis C RNA and hepatitis G RNA, respectively (p = 0.815). All infected patients except one in each group reported a risk factor for viral hepatitis. Individuals with tattoos were more likely to have body piercing (p = 0.049; CI = 1.000-1.995), and more than five sexual partners (p = 0.013; CI = 1.073-1.846) than the control group. CONCLUSION: We find no evidence to support the observation that tattoos serve as a risk factor for chronic viral hepatitis. PMID- 10811346 TI - Prevalence, patterns, and course of past hepatitis B virus infection in intravenous drug users with HIV-1 infection. AB - OBJECTIVE: Hepatitis B virus (HBV) and human immunodeficiency virus (HIV) share common routes of transmission. Therefore, markers of either active or past HBV infection are present in many HIV-infected patients, particularly in intravenous drug users (IDUs). The aim of this study was to analyze the serological pattern of past HBV infection (presence or absence of anti-HBs) and the course of past HBV infection (changes in anti-HBs status, and HBV reactivation) in two cohorts of IDUs with and without HIV infection. METHODS: HBV serum markers were studied in 388 HIV-positive and 197 HIV-negative IDUs. Among them, 263 HIV-positive and 50 HIV-negative patients with past HBV infection (serum HBsAg negative and anti HBc positive, with or without anti-HBs) were followed-up for a median of 21 and 13 months, respectively, to detect changes in anti-HBs status and HBV reactivation. RESULTS: The prevalence of HBV infection (either active or past) was higher in HIV-positive than in HIV-negative cases (90% vs 62%, p < 0.001), even when stratified by years of drug use. Most cases (92% of HIV-positive and 89% of HIV-negative) had markers of past infection. Among those patients with past HBV infection, 60% of HIV-positive and 72% of HIV-negative presented serum anti-HBs (p = 0.03). The incidence of anti-HBs loss was 1.8 cases/100 person-year in HIV-positive, and 1.8 cases/100 person-year in HIV-negative patients (RR 1.0, 95% CI 0.1-94, p = NS). Incidence of anti-HBs development was 17.6 cases/100 person-year in HIV-positive and 25.6 cases/100 person-year in HIV-negative IDUs (RR, 1.5, 95% CI, 0.6-3.5, p = NS). Only one HIV-positive patient with markers of past HBV infection developed an active infection (0.2 events/100 person-year). CONCLUSIONS: HBV infection (either active or past) is particularly frequent in HIV-positive IDUs. Most cases have markers of past infection. Isolated detection of anti-HBc (absence of anti-HBs) is more common in HIV-positive than in HIV negative IDUs. Despite their progressive immunosuppression, both anti-HBs loss and HBV reactivation are rare in HIV-infected IDUs. PMID- 10811347 TI - Cost of treating an episode of variceal bleeding in a VA setting. AB - OBJECTIVES: The specific aims of this study were to develop a demographic description of a sample of patients presenting with bleeding esophageal varices and determine the direct health care costs of variceal bleeding. METHODS: This was a retrospective evaluation of patients who underwent esophagogastroduodenoscopy at the Portland VA Medical Center between January 1993 and May 1997. Data sources included both electronic databases and patient medical charts. The primary unit of analysis was an episode of care, defined as an index bleed plus 6 months of follow-up or death, whichever came first. RESULTS: The total inpatient direct cost was $1,566,904 and outpatient direct cost was $104,611, for a total of $1,671,515 for 100 bleeding episodes in 79 patients. Episodes of care for patients receiving < or =2 units of packed red blood cells were approximately a third as costly as those receiving >2 units of packed red blood cells (n = 17, $6,470 and n = 83, $17,553). The difference in costs was statistically significant (p < 0.05), and primarily attributable to hospital bed costs. CONCLUSIONS: There is a substantial financial burden associated with this illness, primarily attributable to inpatient costs. In addition to severity of bleeding, Child's class, endoscopic findings, and the timing of pharmacological therapy seem to influence the overall cost of managing esophageal varices. PMID- 10811348 TI - A prospective multicenter evaluation of new fecal occult blood tests in patients undergoing colonoscopy. AB - OBJECTIVE: Guaiac-based fecal occult blood (FOB) tests, in particular, Hemoccult II (HO), are commonly used to detect colorectal neoplasia. Because the sensitivity and specificity of these tests are critical to cost-effective screening programs, we aimed to investigate the improved performance characteristics of new FOB tests for known colonic lesions. METHODS: Nine centers worldwide performed FOB testing with guaiac-based tests (Hemoccult II [HO] and Hemoccult II SENSA [SENSA]) and immunochemical tests (HemeSelect [HS] and FlexSure OBT [FS]) on 554 patients referred for colonoscopy for predetermined indications. A combination testing strategy consisting of SENSA followed by HS or FS (which was considered positive only when both tests were positive) was also evaluated. Results of FOB tests were compared to findings on colonoscopy. RESULTS: Cancers were identified in 2.9% of subjects, whereas adenomas > or =10 mm were found in 39 patients. Small adenomas, colitis, and other lesions were identified in 141 patients. The positivity rate of HO for adenomas > or =10 mm was less than for SENSA (20.5% vs 35.9%, p < 0.05), whereas the positivity rate of HO, SENSA, FS, HS, or the combination tests for cancers was not statistically different. The overall positivity rates were significantly greater for FS (15.9%, p = 0.0002) and significantly lower using the combination tests (SENSA/FS 6.0%, p = 0.01; SENSA/HS 6.2%, p = 0.02) compared to HO (9.4%). In this study population, the relative specificity (i.e., true-negative tests/true-negatives + false positives in patients without adenomas > or =10 mm or cancers) of HO (93.9%; 95% CI, 91.7-96.1) was similar to that of SENSA (92.8%; 95% CI, 90.4-95.2) and HS (90.1%; 95% CI, 87.4-92.8), and greater than FS (88.0%; 95% CI, 85.1-90.9, p < 0.001). When considering adenomas > or =10 mm, cancers alone or cancers and adenomas combined, the combination test using SENSA/FS was associated with significantly fewer false-positive tests than any of the individual tests. CONCLUSIONS: Compared to single tests, the combination test with the highly sensitive SENSA and an immunochemical test had slightly reduced sensitivity but significantly fewer false-positive tests than any single test. These data raise the possibility that a combination test (i.e., highly sensitive guaiac plus immunochemical) could reduce the costs of screening for colon cancer, and suggest that further study of combination test strategies is warranted. PMID- 10811349 TI - A look back. Mouth gag. PMID- 10811350 TI - Prolonged cholestasis after acute paraquat poisoning through skin absorption. AB - Ingestion of paraquat is the most common cause of fatal pesticide poisoning. Liver involvement in acute paraquat poisoning is self-limited and usually consists of cholestasis. However, long-term hepatic effects after paraquat exposition have not been described up to now, probably because of the high mortality rate of this acute poisoning. We report the case of an agricultural worker who developed persistent cholestasis after an episode of acute paraquat poisoning through skin absorption. PMID- 10811351 TI - A carcinoembryonic antigen-secreting adenocarcinoma arising within a retrorectal tailgut cyst: clinicopathological considerations. AB - Retrorectal tailgut cysts (TGC) develop from postanal fetal gut remnants. They have specific radiological and histopathological features that distinguish them from dermoid cysts, enteric duplication cysts, and teratomas. We report a patient with a carcinoembryonic antigen-producing adenocarcinoma arising within a TGC who underwent resection through a combined anterior laparotomy/posterior pelvic approach. Despite complete resection and delayed but complete functional recovery, diffusely metastatic disease was encountered 6 months after resection. Diagnostic, therapeutic, histopathological, and oncological implications of this illustrative case are discussed. It seems possible to use carcinoembryonic antigen measurements for treatment planning and for assessing treatment response for this rare disease. The described outcome also suggests that TGC can develop malignant degeneration and should be resected at the time of diagnosis. PMID- 10811352 TI - Ovarian mucinous cystadenocarcinoma as a cause of Zollinger-Ellison syndrome: report of a case and review of the literature. AB - Gastrin producing ovarian tumor is a rare cause of the Zollinger-Ellison syndrome. We report the case of an ovarian carcinoma showing increased plasma gastrin concentration. A 60-yr-old woman presented with epigastric pain and diarrhea. Physical examination showed a large mass in the lower abdomen. Computed tomography revealed a large multilocular ovarian cyst. Upper gastrointestinal endoscopy examination showed multiple ulcerations of the stomach and duodenum. The plasma gastrin level was 1500 pg/ml. No tumors were found in the pancreas and duodenum, and salpingo-oophorectomy was performed. Histologic examination revealed a mucinous tumor of borderline malignant potential. Immunoperoxidase studies for gastrin showed many gastrin-producing cells within the epithelium of the tumor. There have been 11 cases (including our patient) of gastrin-producing ovarian tumor reported in the literature. We review here the relevant literature. Although ovarian gastrinoma is extremely rare, it should be considered as a possible cause of the Zollinger-Ellison syndrome in women. PMID- 10811353 TI - A case of cancerous familial adenomatous polyposis in urinary bladder due to migration of colonic mucosa through rectovesical fistula. AB - The patient was a 50-yr-old man who had undergone low anterior resection for rectal cancer at the age of 24 yr in 1966. At that time, gastric and colonic polyposis were indicated. Postoperative anastomotic dehiscence occurred and, by 1985, a rectovesical fistula had formed. In 1986, when the patient was 44 yr old, he was examined at our hospital for constriction of the rectum due to the rectovesical fistula. Abdominoperineal excision of rectum and surgical closure of the fistula were performed, and the patient was kept under observation because of a diagnosis of familial adenomatous polyposis. In 1988, when the patient was 46 yr old, early ascending colon cancer was discovered and total colectomy was performed. Then, in December, 1991, gross hematuria was found. Further examination revealed a tumor on the posterior wall of the urinary bladder lumen, and biopsy showed adenocarcinoma. Pelvic recurrence of the rectal cancer was diagnosed, and total pelvic exenteration was performed. There were no distant metastases; histologically, the tumor of the bladder was thought to be due to colonic mucosa of familial adenomatous polyposis that had migrated to the bladder lumen via the rectovesical fistula and had become cancerous. PMID- 10811354 TI - Diagnosis of acute right lower quadrant pain and appendicitis by CT scan: do we still need the clinician? PMID- 10811355 TI - Therapeutic face-off: band ligation versus beta blockage for variceal bleeding. PMID- 10811356 TI - Staging rectal carcinoma with MR imaging: improving accuracy with newer techniques. PMID- 10811357 TI - Topical pharyngeal anesthesia for upper gastrointestinal endoscopy. PMID- 10811358 TI - Re: Editorial on outcomes of treatment for GERD. PMID- 10811359 TI - Gastric mucosal lesions in celiac patients. PMID- 10811360 TI - Antiendomysium and antigliadin antibodies for the diagnosis of celiac disease. PMID- 10811362 TI - Screening for hereditary hemochromatosis. PMID- 10811361 TI - An unusual visualization of the gallbladder by somatostatin receptor (SSTR) scintigraphy: usefulness of hepatobiliary imaging for differential diagnosis. PMID- 10811363 TI - Transmission of HCV: the Portuguese case. PMID- 10811364 TI - Relationship between aminotransferase levels and histopathological findings in patients with nonalcoholic steatohepatitis. PMID- 10811365 TI - Inducible nitric oxide synthase activity is expressed not only in inflamed but also in normal colonic mucosa in patients with ulcerative colitis: a potential prognostic marker. PMID- 10811366 TI - Obesity associated with severe acute colonic diverticulitis in the young adult. PMID- 10811367 TI - Re: Pregnancy outcomes in celiac women. PMID- 10811368 TI - Endoscopic ultrasonography in the diagnosis of aortoesophageal fistula. PMID- 10811369 TI - Interobserver variation in describing endoscopic features in inflammatory bowel disease using videotapes of colonoscopy. PMID- 10811370 TI - Treatment of the wrong disease with the right medication: a case of generalized leishmaniasis involving the liver and the gastrointestinal tract. PMID- 10811371 TI - Role of magnetic resonance cholangiopancreatography in diagnosing pancreatic cancer originating from the peripheral duct. PMID- 10811372 TI - Bile peritonitis secondary to breast cancer metastatic to the gallbladder. PMID- 10811373 TI - Transvaginal versus transrectal ultrasonography in the diagnosis of peritoneal carcinomatosis. PMID- 10811374 TI - Can CEA predict malignancy of cystic tumor of the appendix? Report of a case. PMID- 10811375 TI - Severe upper GI bleeding secondary to a migrated J tube: a rare complication. PMID- 10811376 TI - Successful desensitization to 6-mercaptopurine in a patient with Crohn's disease. PMID- 10811377 TI - Unilateral thalamic stroke does not decrease ipsilateral sleep spindles. AB - STUDY OBJECTIVES: To measure the sleep spindle characteristics in patients with unilateral thalamic stroke. DESIGN: A prospective study of patients with thalamic stroke and age-matched healthy controls. SETTING: Department of Neurology of a University Hospital. PARTICIPANTS: Thirteen patients (mean age: 67 years, SD: 13,44) with an isolated, unilateral acute thalamic stroke and 18 healthy age matched volunteers. INTERVENTIONS: A polysomnogram recording from 14 scalp EEG electrodes performed during 2 consecutive nights, the second or third week after the stroke. Only the sleep of the second night was analyzed. MEASUREMENTS AND RESULTS: Sleep spindles were counted during two separate 10-minute epochs of stage II. Spindles appearing synchronously in both sides with similar amplitude were called "bilateral." Spindles with twice the amplitude in one side than the other were "right" or "left-side predominant". There were 8 patients with posterolateral, 3 with global and 2 with anterior lesions. Eight were right and 5 left-sided. The number of spindles was similar in patients (39.8 +/- 23.4 in 20 minutes) than controls (26.07 +/- 29.07; p=0.173). Spindles with a centroparietal (34%) and centroparieto-occipital localization (22%) were the most frequent. In controls approximately 66% of the spindles had a bilateral and symmetric distribution over the scalp, 23% of the spindles were predominantly left-sided and 5% were predominantly right-sided. In patients, bilateral spindles decreased (p<0.0001) but asymmetric spindles did not change. CONCLUSION: Unilateral acute thalamic stroke does not decrease sleep spindles ipsilaterally; rather, it seems to produce a bilateral diminution in their number. PMID- 10811378 TI - Vascular resistance in the rat during baseline, chronic total sleep deprivation, and recovery from total sleep deprivation. AB - Rats subjected to total sleep deprivation (TSD) by the disk-over-water method exhibit an elevated temperature set point, increased energy expenditure (EE), and increased circulating norepinephrine--all of which should militate for an increase in body temperature. Instead, after a small rise early in TSD, intraperitoneal temperature (T(ip)) fell progressively, indicating a reduced ability to retain body heat. To evaluate whether vasoconstrictor defenses against heat loss in the regions of major heat dissipation in the rat (hindpaws and tail) were impaired, peripheral vascular resistance (PVR) was calculated from aortic blood pressure (BP) and blood flow (BF) (BP and BF were continuously recorded at the aortic-iliac junction). TSD rats and their yoked control (TSC) rats were subjected to TSD for 10 to 22 days. As in earlier studies, TSD rats showed excessive heat loss indicated by a falling T(ip) (after an initial rise) while EE was elevated. Temperature set point was presumably raised throughout deprivation as shown previously. Although a small decline in PVR early in deprivation could have increased heat loss, there was no evidence of a massive vasodilation in the region examined which could, in itself, account for the progressive inability to retain heat over the course of TSD. In fact, PVR was near baseline levels during the latter half of TSD. Nevertheless, there was evidence of impaired vasoconstrictive defenses in TSD rats inasmuch as they showed significantly lower PVR than TSC rats during most of the deprivation period in spite of indications that they were farther below set point. It is not yet clear whether this impairment was a major determinant of the heat loss in TSD rats. A rapid PVR rebound during recovery suggested a release from a TSD-linked blockage of vasomotor compensation for excessive heat loss. PMID- 10811379 TI - Treatment of idiopathic restless legs syndrome (RLS) with the D2-agonist cabergoline--an open clinical trial. AB - STUDY OBJECTIVES: To define the effective dose of cabergoline and to evaluate the tolerability and efficacy of cabergoline in patients with restless legs syndrome (RLS). DESIGN: Treatment of idiopathic RLS patients with cabergoline in a 12-week open label trial. Patients on levodopa therapy were allowed to either stop levodopa prior to study entry or to continue, taper or discontinue levodopa during the study. Efficacy parameters were assessed by polysomnography and subjective ratings at baseline and at week 12. Primary efficacy parameters were the number of PLM and total sleep time. SETTING: Dept. of Neurology, Sleep Disorders Center PATIENTS: Nine patients with moderate to severe RLS (age 38.1 to 64.3 years, mean 54.1 years) who had experienced insufficient benefit under levodopa therapy and/or in part developed daytime augmentation participated. At study entry five patients were still under levodopa therapy (400-800 mg). INTERVENTIONS: Up-titration of cabergoline (single evening dose) until RLS symptoms clearly improved. Initial comedication with domperidone 20 mg t.i.d. MEASUREMENTS AND RESULTS: At the endpoint all patients were on cabergoline monotherapy (mean dosage 2.1 mg, range 1 to 4 mg). Domperidone was stopped in all patients due to good tolerability. Polysomnographic data showed a significant reduction of the number of periodic leg movements (PLM) (195.8+/-109.1 (baseline) vs. 26.4+/-40.2 (12 weeks cabergoline monotherapy; p=0.002), PLM arousals (51.7+/ 42.3 vs. 6.4+/-11.2; p=0.017) and PLM awakenings (10.4+/-7.8 vs. 1.0+/-1.7; p=0.001). Total sleep time was prolonged (302.7+/-50.7 vs. 379.4+/-59.8 min; p=0.018), sleep latency shortened (42.4+/-49.1 vs. 16.3+/-22.8 min; p=0.214) and sleep efficiency increased (63.1+/-10.5 vs. 79.1+/-12.5%; p=0.017). All patients reported a impressive relief or became free of RLS symptoms. CONCLUSION: Cabergoline is effective and well tolerated in restless legs syndrome especially in patients with severe RLS and those who developed augmentation under levodopa therapy. PMID- 10811380 TI - Sleep and periodic leg movement patterns in drug-free patients with Parkinson's disease and multiple system atrophy. AB - STUDY OBJECTIVE: To assess and compare polygraphic sleep measures and periodic leg movement (PLM) patterns in untreated patients with mild to moderate Parkinson's disease (PD), multiple system atrophy (MSA) and healthy age-matched controls. DESIGN: Polysomnographic recordings of 2 consecutive nights were performed in 10 patients with PD (mean age 65 years, mean Hoehn and Yahr stage 2.2), 10 patients with MSA (mean age 61 years) and in a control group of 10 healthy subjects (mean age 64 years). All patients and controls were free of antiparkinsonian medication and other centrally active drugs for 2 weeks prior to polysomnography. SETTING: NA. PATIENTS OR PARTICIPANTS: NA. INTERVENTIONS: NA. RESULTS: Sleep measures for the second night showed a significantly lower total sleep time, sleep efficiency and sleep period time in PD and MSA patients compared to healthy controls. PLM-indices during sleep and wakefulness were significantly higher in PD, but not in MSA patients compared to controls. Five patients with PD and 7 patients with MSA, but no control subject, showed abnormal rapid eye movement (REM) sleep features (i.e., REM sleep without atonia or behavioral manifestations typical for REM sleep behavior disorder). CONCLUSIONS: Sleep disruption and increased motor activity during REM and non REM sleep are a frequent finding in PD and MSA. An increased PLM index in untreated PD patients may be due to a dopaminergic deficit and is probably not associated with dopaminergic treatment. PMID- 10811381 TI - Cognitive deficits associated with sleep apnea syndrome: a proposed neuropsychological test battery. AB - Numerous studies have assessed a wide range of cognitive deficits associated with obstructive sleep apnea syndrome (OSAS). The comparison of these various results, however, is hampered by the fact that different studies employ different neuropsychological tests, even when assessing the same function. The aim of this paper is to present a standardized neuropsychological test battery for the evaluation of OSAS patients. Following a description of the general characteristics of OSAS, we review the main cognitive functions reported as being impaired in OSAS patients. These include general intellectual functioning, attentional functioning, memory and learning abilities, executive functions, and motor performance. Based on this review, we propose a test battery designed to cover these cognitive processes while taking into account the time constraints present in most research and clinical centers. In addition to providing a comprehensive neuropsychological evaluation of OSAS patients, the proposed test battery should facilitate the comparison of results from different laboratories. PMID- 10811382 TI - Sleepiness-related accidents in sleep apnea patients. AB - The frequency of motor vehicle and working accidents was analyzed by means of a strictly anonymous questionnaire in 156 patients with sleep apnea syndrome (SAS) and in 160 age-gender matched controls. In the SAS group 12.4% of all drivers had motor vehicle accidents as compared to 2.9% in the control group (p<0.005). The motor vehicle accident rate was 13.0 per million km in patients with more severe SAS (AHI > 34/h, n=78) as compared to 1.1 in patients with milder SAS (AHI 10 34/h, n=78) (p<0.05), and 0.78 in control group (p<0.005), respectively. The accident rates in both patients and the control group were also greater than the rate of 0.02 "accidents due to sleepiness" per one million km in the Swiss driving population as reported by official statistics. During treatment with nasal continuous airway pressure (nCPAP) in 85 SAS patients, the motor vehicle accident rate dropped from 10.6 to 2.7 per million km (p<0.05). We conclude that patients with moderate to severe SAS have an up to fifteen-fold risk increase of motor vehicle accidents that constitutes a serious and often underestimated hazard on the roads, which can be reduced by adequate treatment. PMID- 10811383 TI - Driving simulation with EEG monitoring in normal and obstructive sleep apnea patients. AB - STUDY OBJECTIVES: To measure simulated driving performance in obstructive sleep apnea patients and its relationship with EEG defined attention lapses. DESIGN: Prospective, mixed design comparing apnea patients and control subjects over a 60 minute driving simulation task while continuously recording both driving performance and EEG measures. SETTING: Sleep disorders center. PARTICIPANTS: 15 polysomnographically diagnosed obstructive sleep apnea patients (mean age 42 +/- 6 yrs.) and 15 healthy volunteers (mean age 38 +/- 6 yrs.). INTERVENTIONS: NA. MEASUREMENTS AND RESULTS: A computer based driving simulator recorded lane position variability, speed variability, steering rate variability, and crash frequency. The frequency and duration of EEG-defined attention lapses were also measured. The results demonstrated that the apnea group had significantly greater variability in lane position, steering rate, and speed than the control group. The apnea group also had more crashes. In addition, the apnea group had more EEG defined attention lapses of longer duration. Except for speed and steering rate variability, these differences increased over the 60-minute task. Measures of lane position variability and crash frequency had a significant positive correlation with attention lapse frequency and duration. CONCLUSIONS: The driving simulation task unmasked and quantified marked performance impairments in the sleep apnea group that increased over time. The poor performance appeared related to the EEG-defined attention lapses. Lane position variability appeared to be the most sensitive measure for assessing and quantifying impairment. This study suggests that poorer driving performance and crashes are not entirely due to overt sleep, but inattention due to sleepiness. PMID- 10811384 TI - Influence of ambient temperature on sleep characteristics and autonomic nervous control in healthy infants. AB - OBJECTIVE: To evaluate the influence of ambient temperature on infant's sleep and cardiorespiratory parameters during sleep. PATIENTS AND METHOD: 20 healthy infants with a median age of 11.5 weeks (range 7 to 18 weeks) were recorded polygraphically for one night. They were exposed to 3 different ambient temperatures (20 degrees C-25 degrees C-30 degrees C). Ambient and core temperatures were measured throughout the procedure. RESULTS: Influence of ambient temperature was seen in: RESPIRATORY PARAMETERS: The frequency of central apneas increased significantly with increasing temperatures in REM sleep, but not in NREM sleep. HEART RATE (HR) PARAMETERS AND HR SPECTRAL ANALYSIS: Elevation of temperature was characterized by significantly higher basal HR, shorter RR intervals, and lower parasympathetic activity in REM and NREM sleep. SATURATION IN OXYGEN: During total sleep time, rise in temperature induced a decrease in basal oxygen saturation. During REM sleep, a greater frequency of oxygen saturation drops was associated with central apneas. CORE TEMPERATURE: With increasing ambient temperature, the rise of rectal temperature was mild. Despite this lack of significant increase, similar results were found when sleep and cardiorespiratory parameters were evaluated according to rectal temperatures. CONCLUSION: Changes in ambient temperatures associated with mild increases in body temperature significantly modified cardiorespiratory parameters and autonomic controls in healthy infants. The changes associated with increases in temperature were mainly seen during REM sleep. PMID- 10811385 TI - Effects of a nocturnal environment perceived as warm on subsequent daytime sleep in humans. AB - We studied the influence of a nocturnal environment perceived as warm on the subsequent daytime sleep of healthy human subjects (20-25 years old). From 00:00 to 8:00, they were kept awake and exposed to either a thermoneutral and comfortable (CN) or a warm and uncomfortable (EW) environment, as assessed by the predicted mean vote/percentage of persons dissatisfied questionnaire (PMV/PPD). The subjects then slept from 8:00 to 14:00 in a thermoneutral environment. Sleep was scored according to the Rechtschaffen and Kales criteria. Rectal temperature was recorded from 22:00 to the end of the sleep period. Compared to CN, a significant but moderate hyperthermia (0.3-0.4 degrees C) occurred rapidly in EW, and was maintained throughout the night. This modest difference disappeared during subsequent sleep spent at thermal comfort. Exposure to a warm uncomfortable environment before bedtime significantly increased the duration (+37%) and percentage of rapid eye movement sleep (REMS). This hypnic response could be due to interactions occurring between thermoregulatory, circadian, and sleep mechanisms. It could also be ascribed to synergic actions of the neurophysiological (among others, involving the hypothalamo-pituitary-adrenal axis) and psychological (involving memory processing) processes developed when the organism faces a moderate stress. PMID- 10811386 TI - Factors predicting sleep disruption in Type II diabetes. AB - This study investigated four correlates of diabetes as potential predictors of sleep disruption in people with Type II diabetes. A tentative causal model proposed obesity would lead to more severe diabetes, which would result in increased physical complications, which might in turn affect emotional adjustment, and all of these might combine to cause sleep disruption. It was found that within a diabetic sample, severity of diabetes was related to sleep disruption. Path analysis indicated that the relationship between Type II diabetes and sleep disruption was mediated by physical complications of the disease (pain and nocturia), and not by either obesity or emotional adjustment. PMID- 10811387 TI - Sleep spindle incidence dynamics: a pilot study based on a Markovian analysis. AB - Results are reported, based on 5 healthy subjects, concerning patterns in the dynamics of the sequential arrangement of spindles in human stage 2 sleep. Specifically, the conditional probability of incidence of successive spindle lengths and successive inter-spindle intervals is investigated. The results show that successive spindle lengths may be statistically independent. However, their distribution (histogram) may be similar for two different stage 2 periods, one in the first third and another in the second third of the night sleep record. In contrast to the finding about spindle lengths, results show that successive inter spindle intervals may not be statistically independent. Furthermore, the overall dynamics of the sequential arrangement of inter-spindle intervals may be similar for the two sleep periods. These findings are discussed in the context of the "sleep maintenance" role of spindles. PMID- 10811388 TI - The Network of European CNS Transplantation and Restoration (NECTAR): an introduction on the occasion of its tenth meeting. PMID- 10811389 TI - Neural stem cells: from cell biology to cell replacement. AB - A large number of crippling neurological conditions result from the loss of certain cell populations from the nervous system through disease or injury, and these cells are not intrinsically replaced. Mounting evidence now suggests that replacement of depleted cell populations by transplantation may be of functional benefit in many such diseases. A diverse range of cell populations is vulnerable, and the loss of specific populations results in circumscribed deficits in different conditions. This diversity presents a considerable challenge if cell replacement therapy is to become widely applicable in the clinical domain, because each condition has specific requirements for the phenotype, developmental stage, and number of cells required. An ideal cell for universal application in cell replacement therapy would possess several key properties: it would be highly proliferative, allowing the ex vivo production of large numbers of cells from minimal donor material; it would also remain immature and phenotypically plastic such that it could differentiate into appropriate neural and glial cell types on, or prior to, transplantation. Critically, both proliferation and differentiation would be controllable. This review considers some of the evidence that stem cells exist in the central nervous system and that they may possess characteristics that make them ideal for broad application in cell replacement therapy. PMID- 10811390 TI - Conditionally immortalized, multipotential and multifunctional neural stem cell lines as an approach to clinical transplantation. AB - Experiments are described using rats with two kinds of brain damage and consequent cognitive deficit (in the Morris water maze, three-door runway, and radial maze): 1) ischemic damage to the CA1 hippocampal cell field after four vessel occlusion (4VO), and 2) damage to the forebrain cholinergic projection system by local injection of excitotoxins to the nuclei of origin or prolonged ethanol administration. Cell suspension grafts derived from primary fetal brain tissue display a stringent requirement for homotypical cell replacement in the 4VO model: cells from the embryonic day (E)18-19 CA1 hippocampal subfield, but not from CA3 or dentate gyrus or from E16 basal forebrain (cholinergic rich) led to recovery of cognitive function. After damage to the cholinergic system, conversely, recovery of function was seen with cell suspension grafts from E16 basal forebrain or cholinergic-rich E14 ventral mesencephalon, but not with implants of hippocampal tissue. These two models therefore provided a test of multifunctionality for a clonal line of conditionally immortalized neural stem cells, MHP36, derived from the E14 "immortomouse" hippocampal anlage. Implanted above the damaged CA1 cell field in 4VO-treated adult rats, these cells (multipotential in vitro) migrated to the damaged area, reconstituted the gross morphology of the CA1 pyramidal layer, took up both neuronal and glial phenotypes, and gave rise to cognitive recovery. Similar recovery of function and restoration of species-typical morphology was observed when MHP36 cells were implanted into marmosets with excitotoxic CAI damage. MHP36 implants led to recovery of cognitive function also in two experiments with rats with excitotoxic damage to the cholinergic system damage, either unilaterally in the nucleus basalis or bilaterally in both the nucleus basalis and the medial septal area. Thus, MHP36 cells are both multipotent (able to take up multiple cellular phenotypes) and multifunctional (able to repair diverse types of brain damage). PMID- 10811391 TI - The use of adenoviral vectors and ex vivo transduced neurotransplants: towards promotion of neuroregeneration. AB - Regeneration of injured axons following injury depends on a delicate balance between growth-promoting and growth-inhibiting factors. Overexpression of neurotrophin genes seems a promising strategy to promote regeneration. Trophic genes can be overexpressed at the site of injury at the axonal stumps, or at the perikaryal level of the injured neuron. Transduction of the neural cells can be achieved by applying adenoviral vectors, either directly in vivo or-in the case of neurotransplantation as an ex vivo approach. In both cases it would create a more permissive environment for axonal growth and therefore in functional regeneration. In this article, the feasibility of the use of adenoviral vectors in several neuroregeneration models--in particularly in spinal cord lesion models and the biological clock transplantation model--is illustrated. The results show that the adenoviral vectors can be a powerful tool to study the effects of overexpression of genes in an in vivo paradigm of nerve regeneration or nerve outgrowth. The potential use of adenoviral vectors and ex vivo transduced neurotransplants is discussed. PMID- 10811392 TI - Improving the survival of grafted dopaminergic neurons: a review over current approaches. AB - Neural transplantation is developing into a therapeutic alternative in Parkinson's disease. A major limiting factor is that only 3-20% of grafted dopamine neurons survive the procedure. Recent advances regarding how and when the neurons die indicate that events preceding actual tissue implantation and during the first week thereafter are crucial, and that apoptosis plays a pivotal role. Triggers that may initiate neuronal death in grafts include donor tissue hypoxia and hypoglycemia, mechanical trauma, free radicals, growth factor deprivation, and excessive extracellular concentrations of excitatory amino acids in the host brain. Four distinct phases during grafting that can involve cell death have been identified: retrieval of the embryo; dissection and preparation of the donor tissue; implantation procedure followed by the immediate period after graft injection; and later stages of graft maturation. During these phases, cell death processes involving free radicals and caspase activation (leading to apoptosis) may be triggered, possibly involving an increase in intracellular calcium. We review different approaches that reduce cell death and increase survival of grafted neurons, typically by a factor of 2-4. For example, changes in transplantation procedure such as improved media and implantation technique can be beneficial. Calcium channel antagonists such as nimodipine and flunarizine improve nigral graft survival. Agents that counteract oxidative stress and its consequences, such as superoxide dismutase overexpression, and lazaroids can significantly increase the survival of transplanted dopamine neurons. Also, the inhibition of apoptosis by a caspase inhibitor has marked positive effects. Finally, basic fibroblast growth factor and members of the transforming growth factor-beta superfamily, such as glial cell line-derived neurotrophic factor, significantly improve the outcome of nigral transplants. These recent advances provide hope for improved survival of transplanted neurons in patients with Parkinson's disease, reducing the need for human embryonic donor tissue and increasing the likelihood of a successful outcome. PMID- 10811393 TI - Complex sensorimotor behavioral changes after terminal striatal 6-OHDA lesion and transplantation of dopaminergic embryonic micrografts. AB - In this study sensorimotor behavioral changes were monitored in rats following bilateral 6-hydroxydopamine (6-OHDA) axon terminal lesion and uni- or bilateral implantation of embryonic dopaminergic (DA) micrografts. A total of 28 microg of 6-OHDA was distributed over four injection tracts in the dorsolateral part of the caudate-putamen (CPU) bilaterally followed 4 months later by the implantation of DA micrografts spread over seven implantation tracts placed within the denervated area. Bilaterally 6-OHDA-lesioned animals exhibited significantly reduced behavioral performance scores in tests of explorational and stepping behavior as well as in skilled forelimb use. However, in contrast to the established medial forebrain bundle (MFB) lesion model of PD, these animals showed a spontaneous recovery in the side falling and skilled forelimb behavior and no deficits in overnight locomotor activity at 6 months after the lesion. Unilateral DA micrografts elicited a substantial amphetamine-induced rotational bias contralateral to the graft, but led to a significant impairment of contralateral skilled forelimb use and reduced scores in overnight locomotor activity. Bilateral DA micrografts caused a significant, though partial, increase in explorational and backhand stepping behavior, but resulted also in a significant decrease in performance levels in overnight locomotor activity and skilled forelimb use on both paws. In conclusion, DA grafts placed ectopically in the CPU in the partial lesion model of PD result in a double innervation of the GABAergic striatal neurons, arising from the residual nigrostriatal DA projections of the host and from the graft-derived DA efferent fibers. These two DA fiber systems may indeed function in a cooperative and competitive manner depending on their respective and different afferent and efferent connections, which, in turn, may lead to positive or negative influences on basal ganglia function and behavioral performances. The different patterns of 6-OHDA lesion and transplant-induced behavioral changes demonstrated in the present study compared to the "classical" MFB lesion model of PD may thus provide further insights in the complex functional organization of the basal ganglia and, thereby, may help to further optimize restorative strategies for neurodegenerative diseases, such as Parkinson's disease. PMID- 10811394 TI - Implication of the subthalamic nucleus in the pathophysiology and pathogenesis of Parkinson's disease. AB - The subthalamic nucleus (STN) has been shown to play an important role in the control of movement and has been considered as a key structure in the functional organization of the basal ganglia. Several studies postulated that the STN plays a critical role in the pathophysiology of Parkinson's disease and that its inhibition or its lesioning can reverse the cardinal motor symptoms. Nevertheless, the beneficial effect was accompanied by dyskinetic abnormal movements. In order to avoid unpleasant and irreversible side effects we used high-frequency stimulation (HFS) of the STN instead of lesions. We have shown that parkinsonian motor symptoms, akinesia, rigidity, and tremor can be alleviated by HFS of the STN in the nonhuman primate model. Side effects were controllable and appeared only at intensities higher than that inducing the improvement of motor symptoms. In severe parkinsonian patients, bilateral STN-HFS greatly improved parkinsonian motor symptoms. Motor fluctuations were attenuated and patients became independent in most activities of daily living. It appears that STN-HFS mimics the effects of lesions by inhibiting its neuronal activity. In a rat model of parkinsonism, we studied the implication of the STN in the excitotoxicity of nigral dopamine cells. We showed that kainic acid lesioning of the STN can protect nigral dopaminergic cells against 6-hydroxydopamine-induced toxicity. The evidence reviewed in the present article clearly demonstrates that the STN is implicated in the pathophysiology and pathogenesis of Parkinson's disease. PMID- 10811395 TI - Towards a protocol for the preparation and delivery of striatal tissue for clinical trials of transplantation in Huntington's disease. AB - There is a growing body of scientific evidence contributing to the development of clinical transplantation programs in patients with Huntington's disease. Phase I clinical trials have already commenced in France and North America and are starting in the near future in Sweden and the UK. Protocols for patient selection, surgical implantation, and pre- and postoperative follow-up are well defined. However, considerable variability exists with respect to the harvesting, preparation, and timing of implantation of the donor material. In this article we review the scientific evidence on which a rational protocol for donor tissue preparation and delivery may be based. Strategies aimed at minimizing the variability of tissue preparation should reduce the variability of functional outcome of striatal transplantation observed in animal models of Huntington's disease. PMID- 10811396 TI - Neural tissue xenotransplantation: what is needed prior to clinical trials in Parkinson's disease? Neural Tissue Xenografting Project. AB - Embryonic allografted human tissue in patients with Parkinson's disease has been shown to survive and ameliorate many of the symptoms of this disease. Despite this success, the practical problems of using this tissue coupled to the ethical restrictions of using aborted human fetal tissue have lead to an exploration for alternative sources of suitable material for grafting, including xenogeneic embryonic dopaminergic-rich neural tissue. Nevertheless, xenografted neural tissue itself generates a number of practical, ethical, safety, and immunological issues that have to be addressed prior to any clinical xenotransplant program. In this article we review these critical issues and set out the criteria that we consider need to be met in the development of our clinical xenotransplantation research programs. We advocate that these, or similar, criteria should be adopted and made explicit by other centers contemplating similar clinical trials. PMID- 10811398 TI - Intrastriatal ventral mesencephalic xenografts of porcine tissue in rats: immune responses and functional effects. AB - Transplantation of neural tissue from other species has the potential to improve function in patients with neurodegenerative disorders. We investigated the functional effects of embryonic porcine dopaminergic neurons transplanted in a rat model of Parkinson's disease and the immune responses to the grafts in immunosuppressed and nonimmunosuppressed hosts. Twenty-three rats with unilateral 6-hydroxydopamine lesions received dissociated, 27-day-old embryonic porcine ventral mesencephalic tissue in the right striatum. Eighteen rats received cyclosporine (10 mg/kg, IP, daily) during the whole period of 14 weeks, in combination with prednisolone (20 mg/kg, IP, daily) the first 4 days. Five rats served as nonimmunosuppressed controls. All rats were tested for amphetamine induced rotational behavior at 3-week intervals. Two immunosuppressed rats were excluded due to severe side effects of the treatment. Functional recovery was seen in 9 of 16 immunosuppressed rats at 12 weeks. Six animals remained functionally recovered at 14 weeks and contained an average of 5750+/-1450 (SEM) dopaminergic neurons. Between 9 and 14 weeks, three immunosuppressed rats rejected their grafts, based on rotation scores and immunohistochemical demonstration of cell infiltrates. One additional immunosuppressed rat showed evidence of ongoing rejection at 14 weeks. The striata in animals with ongoing or recent rejection contained large numbers of CD4- and CD8-positive lymphocytes, NK cells, macrophages, and microglia cells, whereas scar tissue was found in rats with grafts rejected at earlier time points (n = 11). Embryonic porcine ventral mesencephalic tissue matures in the adult rat striatum, reinnervates the host brain, and restores behavioral defects. Immunosuppressive treatment was necessary for long-term graft survival and functional recovery, but did not sufficiently protect from rejection mechanisms. Porcine neural tissue is an interesting alternative to embryonic human tissue for intracerebral transplantation in neurodegenerative diseases. However, to achieve stable graft survival in discordant xenogeneic combinations, an appropriate immunosuppressive treatment or donor tissue modifications are needed. PMID- 10811397 TI - The DaNeX study of embryonic mesencephalic, dopaminergic tissue grafted to a minipig model of Parkinson's disease: preliminary findings of effect of MPTP poisoning on striatal dopaminergic markers. AB - A multicenter study is under way to investigate the efficacy of allografting of embryonic mesencephalic neurons in a pig model of Parkinson's disease. We have first established that a stable parkinsonian syndrome can be established by 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intoxication of adult male Gottingen minipigs. We are now using positron emission tomography (PET) methods for testing the physiological responses to MPTP intoxication and the time course of the response to several treatment strategies. We now report preliminary results obtained in 11 pigs employed in the initial phase of the study; the completed study shall ultimately include 30 pigs. Animals were randomly assigned to one of five groups: 1) Control, 2) MPTP intoxication, 3) MPTP intoxication followed by allograft, 4) MPTP intoxication followed by allograft with immunosuppression, and 5) MPTP intoxication followed by allograft with immunosuppression and co-grafting of immortalized HiB5 cells, which had been manipulated to secrete glia cell line-derived neurotrophic factor (GDNF) (approximately 2 ng GDNF/h/10(5) cells). MPTP was administered (1 mg/kg/day, SC) for 7-10 days until the pigs had developed mild parkinsonian symptoms of muscle rigidity, hypokinesia, and impaired coordination, especially of the hind limbs. Approximately 2 weeks after the last MPTP dose, animals received a T1-weighted magnetic resonance imaging (MRI) scan, and a series of dynamic PET recordings. After the first series of PET scans, four grafts of porcine embryonic mesencephalic tissue (E28 days) were placed in each striatum of some MPTP intoxicated pigs, using MRI-based stereotactic techniques. Immunosuppression of some animals with cyclosporin and prednisolone began just prior to surgery. Two more series of PET scans were performed at 4-month intervals after surgery. After the last scans, pigs were killed and the brains were perfused for unbiased stereological examination of cytological and histochemical markers in striatum and substantial nigra. The behavioral impairment of the animals (the "Parkinson's score") had been evaluated throughout the 8-month period. Kinetic analysis of the first set of PET scans has indicated that the rate constant for the decarboxylation of FDOPA in catecholamine fibers was reduced by 33% in striatum of the mildly parkinsonian pigs. The rate of association of [11C]NS-2214 to catecholamine uptake sites was reduced by 62% in the same groups of pigs. No significant difference was found in the binding potential of [11C]raclopride to the dopamine D2-like receptors in striatum of the MPTP-intoxicated versus control pigs. These preliminary results are suggestive that the activity of DOPA decarboxylase may be upregulated in the partially denervated pig striatum. PMID- 10811399 TI - Porcine xenografts in Parkinson's disease and Huntington's disease patients: preliminary results. AB - The observation that fetal neurons are able to survive and function when transplanted into the adult brain fostered the development of cellular therapy as a promising approach to achieve neuronal replacement for treatment of diseases of the adult central nervous system. This approach has been demonstrated to be efficacious in patients with Parkinson's disease after transplantation of human fetal neurons. The use of human fetal tissue is limited by ethical, infectious, regulatory, and practical concerns. Other mammalian fetal neural tissue could serve as an alternative cell source. Pigs are a reasonable source of fetal neuronal tissue because of their brain size, large litters, and the extensive experience in rearing them in captivity under controlled conditions. In Phase I studies porcine fetal neural cells grafted unilaterally into Parkinson's disease (PD) and Huntington's disease (HD) patients are being evaluated for safety and efficacy. Clinical improvement of 19% has been observed in the Unified Parkinson's Disease Rating Scale "off" state scores in 10 PD patients assessed 12 months after unilateral striatal transplantation of 12 million fetal porcine ventral mesencephalic (VM) cells. Several patients have improved more than 30%. In a single autopsied PD patient some porcine fetal VM cells were observed to survive 7 months after transplantation. Twelve HD patients have shown a favorable safety profile and no change in total functional capacity score 1 year after unilateral striatal placement of up to 24 million fetal porcine striatal cells. Xenotransplantation of fetal porcine neurons is a promising approach to delivery of healthy neurons to the CNS. The major challenges to the successful use of xenogeneic fetal neuronal cells in neurodegenerative diseases appear to be minimizing immune-mediated rejection, management of the risk of xenotic (cross species) infections, and the accurate assessment of clinical outcome of diseases that are slowly progressive. PMID- 10811400 TI - Towards neurotransplantation in multiple system atrophy: clinical rationale, pathophysiological basis, and preliminary experimental evidence. AB - Multiple system atrophy (MSA) is a neurodegenerative disorder that occurs sporadically and causes parkinsonism, cerebellar, autonomic, urinary, and pyramidal dysfunction in many combinations. Progressive L-dopa-unresponsive parkinsonism due to underlying striatonigral degeneration dominates the clinical syndrome in the majority of cases (MSA-P subtype). MSA-P is characterized pathologically by degenerative changes in somatotopically related areas of the substantia nigra pars compacta and of the putamen. Furthermore, oligodendroglial cytoplasmic inclusions (GCIs) are observed throughout the cortico-striato pallidocortical loops and may contribute to the basal ganglia dysfunction. Neurotransplantation strategies are of potential interest in this disease, which causes marked and early disability and dramatically reduces life expectancy. A number of experimental MSA-P models have been employed to evaluate neurotransplantation approaches. Sequential nigral and striatal lesions using 6 hydroxydopamine and quinolinic acid (double toxin-double lesion approach) indicate that apomorphine-induced contralateral rotation is abolished by a secondary striatal lesion. Intrastriatal injection of mitochondrial respiratory chain toxins produces secondary excitotoxic striatal lesions combined with retrograde nigral degeneration and therefore provides an alternative single toxin double lesion approach. Neurotransplantation in MSA-P animal models has been used to improve functional deficits by replacing lost nigral and/or striatal circuitry (neuroregenerative approach). The available data indicate that embryonic mesencephalic grafts alone or combined with striatal grafts partially reverse drug-induced rotation asymmetries without improving deficits of complex motor function. The potential neuroprotective efficacy of embryonic striatal grafts against striatal excitotoxicity is presently under investigation in the double toxin-double lesion MSA-P rat model. Anecdotal clinical evidence in one MSA-P patient misdiagnosed as Parkinson's disease indicates that embryonic mesencephalic grafts produce incomplete clinical benefit. Striatal co-grafts may increase functional improvement. Further experimental studies are required prior to the clinical application of embryonic neurotransplantation in MSA-P. Future research strategies should explore the effect of neurotransplantation in partial MSA-P rat models with less severe nigral and striatal degeneration, the feasibility of a primate model closely mimicking the human disease, and the replication of oligodendroglial dysfunction. PMID- 10811401 TI - Transplantation options for therapeutic central nervous system remyelination. AB - Persistent demyelination, in addition to being the major pathology of multiple sclerosis and the leucodystrophies, is also a feature of spinal cord trauma where there is evidence that it contributes to the functional deficit. In experimental animals it is possible to remyelinate demyelinated CNS axons by transplanting cultures containing central or peripheral myelinogenic cells. Using functional testing we have been able to show that transplant-mediated remyelination results in restoration of function lost as a consequence of demyelination. Glial cell transplantation may therefore provide a therapeutic strategy for remyelinating areas of chronic demyelination. This article reviews issues that have to be addressed before glial transplantation can be undertaken in humans. These include: what cells to use, where would the cells come from, and can we predict how much remyelination will be achieved? It concludes that the most promising approach will be to use neural multipotential stem cells isolated from embryonic CNS, expanded in vitro as neurospheres and then committed to oligodendrocyte lineage differentiation prior to implantation. However, even with such preparations, which have considerable myelinating potential, the extent of remyelination that will be achieved cannot currently be predicted with any degree of certainty. PMID- 10811402 TI - Ultrasonographic evaluation of focal hepatic lesions: comparison of pulse inversion harmonic, tissue harmonic, and conventional imaging techniques. AB - A prospective study was performed to evaluate whether pulse inversion harmonic imaging and tissue harmonic imaging improve the lesion conspicuity and change ultrasonographic characteristics of focal hepatic lesions. Three radiologists evaluated 97 focal hepatic lesions by consensus: cirrhosis-related nodules (n = 23), metastases (n = 23), hemangiomas (n = 27), and cysts (n = 24). In our study, pulse inversion harmonic imaging was judged superior to both tissue harmonic imaging and conventional imaging in conspicuity and overall quality for cirrhosis related nodules, metastases, and cysts (P < 0.05). Compared with conventional imaging, both pulse inversion harmonic imaging and tissue harmonic imaging provided better conspicuity, clearer internal echogenicity, and stronger through transmission of cysts (P < 0.05). Pulse inversion harmonic imaging was judged to be better in depicting internal morphology for cirrhosis-related nodules and metastases than conventional imaging (P < 0.05). For hemangiomas, no statistically significant difference was found in all parameters except better posterior enhancement by tissue harmonic imaging than other techniques. In conclusion, pulse inversion harmonic imaging showed the best conspicuity and also enhanced characteristics of both cystic and solid hepatic lesions. Tissue harmonic imaging was judged superior to conventional imaging in evaluating cysts but was not beneficial for solid lesions. The results of this trial may be specific to the machine used for this study. PMID- 10811403 TI - Understanding the Doppler RI: impact of renal arterial distensibility on the RI in a hydronephrotic ex vivo rabbit kidney model. AB - The aim of this study was to evaluate the influence of elevated ureteral pressure on renal arterial distention, and thereby on the Doppler resistive index. Seven isolated rabbit kidneys were subjected to a pulsatile perfusion while the renal pelvis was pressurized via the ureter. Renal vascular pressure, flow, resistance (pressure/flow) and conductance (flow/pressure) were compared to simultaneous resistive index measurements using linear regression analysis. Changes in the Conductance Index (systolic conductance - diastolic conductance/systolic conductance) were likewise compared to the resistive index. Elevations in ureteral pressure were significantly correlated with (1) increased resistive index values, (2) increased mean renal vascular resistance, (3) decreased mean conductance, and (4) increased conductance index values (for all correlations P < 0.05). The increases in the resistive index correlated significantly with increases in the conductance index. This study shows that elevated ureteral pressure, likely acting via interstitial pressure, diminishes the conductance of the renal vascular bed. Because this effect is more dramatic at diastole, the cyclic patterns of flow are altered, resulting in elevated resistive index values. The results indicate the importance of the interaction between vascular distensibility and pulsatile flow, rather than overall mean renal vascular resistance, in determining resistive index values. PMID- 10811404 TI - Intestinal ultrasonography in children and young adults: bowel wall thickness is age dependent. AB - The purpose of this study was to investigate bowel wall thickness in children, adolescents, and young adults as measured by ultrasonography. Ultrasonographic evaluation of the small bowel and the colon was performed in 128 subjects (age range, 3 days to 40 years). Bowel wall thickness increased significantly (P < 0.0001) increased with age, reaching a maximum colon wall thickness of 2.0 mm at the age range of 20 to 29 years. In the ileum, maximum wall thickness reached 1.5 mm at the age range of 10 to 14 years. The findings emphasize the need for age specific normal values for bowel wall thickness measured by ultrasonography. PMID- 10811405 TI - Shear rate estimation using a clinical ultrasound scanner. AB - Wall shear stress is a factor in the development of atherogenesis, thrombus formation, and embolization, but its existence is very difficult to determine with ultrasonography. Instead, we estimated shear rates using a clinical ultrasound scanner and compared results from vessels with and without stenoses. Velocity profiles were obtained from color M-mode images on a P700 scanner. Maximum shear rates were calculated off-line as the maximum velocity gradients. In vitro, studies were performed on a flow phantom with a vessel containing a 50% stenosis. Shear rates within the stenosis were significantly higher than those obtained outside the stenosis (P < 0.00001) and varied more than the peak velocities. In vivo, the internal carotid artery of 10 volunteers and 13 patients (with stenoses) was studied. The mean shear rate was 414 s(-1) +/- 154.5 s(-1) in normal vessels and 687 s(-1) +/- 263.5 s(-1) in stenotic vessels (P = 0.00017). In conclusion, shear rate estimates can be obtained with a clinical ultrasound scanner from color M-mode images. Shear rates estimated in vessels with stenoses are significantly higher than those obtained in normal vessels in vitro as well as in vivo. PMID- 10811406 TI - Successful ultrasonographically guided compression repair of a dialysis fistula pseudoaneurysm. PMID- 10811407 TI - Sonographic demonstration, including color Doppler imaging, of recurrent sperm granuloma. PMID- 10811408 TI - Prenatal ultrasonographic diagnosis of congenital oral granular cell myoblastoma. PMID- 10811409 TI - Prenatal diagnosis of bilateral fibular hemimelia. PMID- 10811410 TI - Abdominal ultrasonography of splenogonadal fusion. PMID- 10811411 TI - Prenatal diagnosis of congenital pancreatic cyst: case report and review of the literature. PMID- 10811412 TI - Case 1: abdominal. Page kidney. PMID- 10811413 TI - Case 2: abdominal. Testicular microlithiasis complicated by testicular carcinoma. PMID- 10811414 TI - Case 3: obstetrical. Heterotopic pregnancy: viable twin intrauterine pregnancy with a viable right tubal ectopic pregnancy. PMID- 10811415 TI - Case 4: obstetrical. Pulmonary sequestration. PMID- 10811416 TI - Case 5: gynecological. Granulosa cell tumor of the ovary with endometrial carcinoma. PMID- 10811417 TI - Case 6: vascular. Transplant renal vein stenosis. PMID- 10811418 TI - Case 7: obstetrical. Acardiac twinning. PMID- 10811419 TI - Case 8: obstetrical. Lymphangioma of the neck. PMID- 10811420 TI - Case 9: vascular. Tricuspid regurgitation. PMID- 10811421 TI - Case 10: pediatric. Mineralizing (lenticulostriate) vasculopathy. PMID- 10811422 TI - Case 11: obstetrical. Fetal trisomy 18. PMID- 10811423 TI - Case 12: obstetrical. Congenital cytomegalovirus infection. PMID- 10811424 TI - 13C-14N REAPDOR and 13C-2D theta-REDOR NMR on a blend of tri-p-tolylamine and bisphenol-A-polycarbonate. AB - Solid-state NMR experiments are performed on amorphous blends of tri-p-tolylamine (TTA) and polycarbonate. Amines that are similar to TTA are blended with polycarbonates and used as photoconducting components in xerography. Because of its simple molecular structure, TTA is a charge-transporting molecule that is used as a model to understand charge transport in photoconducting materials. Charge transfer between amine molecules must depend on the separation and relative orientation between amine molecules. However, little information is known about the structural relationship between the amine molecules and the polymer host, presumably because of the difficulties associated with characterizing amorphous materials. We report the results of 13C-14N rotational echo, adiabatic-passage, double-resonance (REAPDOR) and 13C-2D theta-rotational echo, double-resonance (theta-REDOR) magic-angle spinning NMR experiments on a TTA/polycarbonate blend. The experiments provide information about the organization of TTA relative to polycarbonate. PMID- 10811425 TI - Non-bridging oxygens in borate glasses: characterization by 11B and 17O MAS and 3QMAS NMR. AB - The concentrations of non-bridging oxygens (NBO) in oxide glasses has major effects on their properties and on those of their precursor glass melts. In borate and borosilicate glasses, the presence of NBO bonded to boron has generally been inferred from 11B NMR spectra and mass balance considerations. Here we report the direct observation of such NBO using 17O MAS and 3QMAS techniques, and compare estimates of their populations with those derived from high-resolution 11B MAS spectra. For the latter, two independent methods are used, based on the ratios of trigonal to tetrahedral boron and on the concentrations of trigonal boron sites with large quadrupolar asymmetry parameters. We include data on crystalline sodium pyroborate (Na4B2O5) and sodium metaborate (NaBO2), and several sodium and barium borate glasses. 17O chemical shifts and quadrupolar coupling constants for NBO bonded to boron vary considerably depending on their coordination environment. In borosilicates, peaks for this species may be hidden by overlap with B-O-Si or Si-O-Si resonances. PMID- 10811426 TI - 2H NMR and polyelectrolyte-induced domains in lipid bilayers. AB - 2H NMR studies of polyelectrolyte-induced domain formation in lipid bilayer membranes are reviewed. The 2H NMR spectrum of choline-deuterated phosphatidylcholine (PC) reports on any and all sources of lipid bilayer surface charge, since these produce a conformation change in the choline head group of PC, manifest as a change in the 2H NMR quadrupolar splitting. In addition, homogeneous and inhomogeneous surface charge distributions are differentiated. Adding polyelectrolytes to lipid bilayers consisting of mixtures of oppositely charged and zwitterionic lipids produces 2H NMR spectra which are superpositions of two Pake sub-spectra: one corresponding to a polyelectrolyte-bound lipid population and the other to a polyelectrolyte-free lipid population. Quantitative analysis of the quadrupolar splittings and spectral intensities of the two sub spectra indicate that the polyelectrolyte-bound populations is enriched with oppositely charged lipid, while the polyelectrolyte-free lipid population is correspondingly depleted. The same domain-segregation effect is produced whether cationic polyelectrolytes are added to anionic lipid bilayers or anionic polyelectrolytes are added to cationic lipid bilayers. The 2H NMR spectra permit a complete characterization of domain composition and size. The anion:cation ratio within the domains is always stoichiometric, as expected for a process driven by Coulombic interactions. The zwitterionic lipid content of the domains is always statistical, reflecting the systems tendency to minimize the entropic cost of demixing charged lipids into domains. Domain formation is observed even with rather short polyelectrolytes, suggesting that individual polyelectrolyte chains aggregate at the surface to form "superdomains". Overall, the polyelectrolyte bound at the lipid bilayer surface appears to lie flat along the surface and to be essentially immobilized through its multiple electrostatic contacts. PMID- 10811427 TI - NMR studies of structure and dynamics in fruit cuticle polyesters. AB - Cutin and suberin are support polymers involved in waterproofing the leaves and fruits of higher plants, regulating the flow of nutrients among various plant organs, and minimizing the deleterious impact of microbial pathogens. Despite the complexity and intractable nature of these plant biopolyesters, their molecular structure and development are amenable to study by suitable solid-state and solution-state NMR techniques. Interactions of tomato cutin with water were examined by solid-state 2H and 13C NMR, showing that water films enhance rapid segmental motions of the acyl chains and are associated with a fivefold increase in surface elasticity upon cutin hydration. The suberization of wounded potato tissues was studied by solid-state 13C NMR, revealing the likely phenylpropanoid structures that permit dense cross-linking of the suberin structure and their proximity to the cell-wall polysaccharides. Finally, two new approaches were developed to elucidate the molecular structures of these biopolymers: partial depolymerization followed by spectroscopic analysis of the soluble oligomers; and swelling of the intact materials followed by magic-angle spinning (MAS) NMR analysis. PMID- 10811428 TI - Fast magic-angle spinning proton NMR studies of polymers at surfaces and interfaces. AB - Solid-state proton NMR with fast magic-angle sample spinning has been used to study the structure and dynamics of polymers and the water interface in porous glass composites. The composites were prepared by photopolymerization of poly(ethyl acrylate) and other acrylate formulations in a high surface-area rigid glass matrix with 40-A interconnected pores. High resolution solid-state proton spectra were obtained for polymer films and composites with 15 kHz magic-angle sample spinning at temperatures above the polymer glass transition temperature. The solid-state proton spectra can be detected with high sensitivity and used to determine the composition of polymer and water filling the pores. These results and spin diffusion studies using 1H-29Si 2D heteronuclear correlation and wideline separation NMR show that the polymer fills the central 30 A of the pore, and that the remaining volume is filled with surface hydroxyl groups and water. PMID- 10811429 TI - Factors influencing the 17O quadrupole coupling constant in bridging oxygen environments. AB - Ab initio calculations were performed on the series of clusters [(OH)3M-O M(OH)3](-2) where M = B, Al, and Ga, (OH)3M-O-M(OH)3 where M = C, Si, and Ge, (OH)2OM-O-MO(OH)2 where M = P and As, and (OH)O2M-O-MO2(OH) where M = S and Se. These clusters were constructed to model the effect of changing the coordinating cation and coordinating cation-oxygen distances on the bridging oxygen 17O quadrupole coupling constant. For all clusters studied, we observed a general trend that the magnitude of Cq increases linearly with increasing cation-oxygen bond distance and cation group number, suggesting that group number and cation bridging oxygen distance can serve as a better predictor of the bridging oxygen quadrupole coupling constant than electronegativity differences. PMID- 10811430 TI - 27Al field-swept and frequency-stepped NMR for sites with large quadrupole coupling constants. AB - Spectra of nonspinning samples with large quadrupole coupling constants, 16-32 MHz, are acquired by frequency-stepping. A series of spin-echoes are acquired at arbitrary frequency increments, frequency-shifted in the time domain, and co added as magnitude spectra. This procedure is derived from a method in use for field-swept NMR. The two methods are compared. PMID- 10811431 TI - Search for retrogressive reactions accompanying demineralization in native and air-oxidized coals. AB - This solid state 13C NMR study confirms, for coals, and extends, for air-oxidized coals, the claim that the Bishop and Ward strong aqueous-acid coal demineralization procedure [M. Bishop, D.L. Ward, Fuel 37 (1958) 191.] does not lead to the formation of detectable levels of cross-links in the organic matrix of these materials. The study was prompted by the demonstration that polymerization accompanies strong acid demineralization in lignin, and the recognition that the chemical environment created in air-oxidized coals contains introduced reactive functionality similar to that in the lignin. In particular, both contain oxidized benzylic carbon functionality that can undergo acid catalyzed polymerization reactions. For two coals of differing rank, no acid induced cross-link formation has been observed in the pristine or air-oxidized coals, within the 5 parts per thousand (ppt) sensitivity of the NMR technique used in this study. PMID- 10811432 TI - Anisotropy-correlated spectroscopy of quadrupolar nuclei. AB - The two-dimensional anisotropy-correlated NMR (2DAC) spectra of half-integer quadrupolar nuclei may be recorded by using an exchange sequence in conjunction with magic angle spinning (MAS) during evolution and detection, and off-MAS during mixing. Application of this experiment to boron oxides is described, in addition to an analysis of the spin diffusion rates in such materials. PMID- 10811433 TI - Identification of fluorine sites at the surface of fluorinated gamma-alumina by two-dimensional MAS NMR. AB - By means of 27Al triple quantum Magic-Angle Spinning Nuclear Magnetic Resonance (3QMAS NMR) and 27Al[19F] WISE MAS NMR, we were able to detect three different Al F sites on the surface of fluorinated gamma-alumina. Three 19F resonances at 9, 20, and 33 ppm (from C6F6) correlated to 27Al resonances in the octahedral range. While the positions of the maxima in the 27Al dimension were ill-defined due to the inherently low efficiency of the 27Al[19F] CPMAS process, the center of gravity of the lines shifted significantly upfield in that dimension with increasing wt.% F. Tentatively, these three resonances were assigned to (VI)Al(O(6-n)Fn) (n = 1, 2, 3) environments on the F/gamma-Al2O3 surface. At F contents above levels corresponding to the full fluorination of the gamma-Al2O3 surface, neoformation of an AlF3 x 3H2O phase was also evidenced with an 19F resonance at -8 ppm and with an 27Al resonance at -17 ppm. PMID- 10811434 TI - Study of the conversion of methanol to dimethyl ether on zeolite HZSM-5 using in situ flow MAS NMR. AB - The conversion of methanol to gasoline (MTG) range hydrocarbons on zeolite catalyst HZSM-5 has been studied extensively using solid-state NMR. We have studied the reaction under batch and flow conditions using an isolated flow variable-temperature (VT) MAS NMR probe. This probe was developed to study heterogeneous catalysis reactions in situ at temperatures greater than 300 degrees C with reactant flow. In the batch studies, when 13C-labeled methanol was adsorbed on zeolite HZSM-5, sealed, and heated to 250 degrees C, dimethyl ether was formed. Two-dimensional exchange NMR shows that dimethyl ether was in equilibrium with methanol at 250 degrees C. When 13C-methanol was flowed over HZSM-5 at temperatures > or = 200 degrees C, only dimethyl ether was observed. Between 160 degrees C and 200 degrees C, both methanol and dimethyl ether were observed. The flow results are significant in that they suggest that there is no equilibrium between methanol and dimethyl ether in the catalyst at high temperatures, and that surface methoxy groups do not exist on the catalyst at high temperatures. PMID- 10811435 TI - Characterization of an Al-, Ga-based catalyst by Ga NMR and XAS. AB - Both Nuclear Magnetic Resonance (NMR) and X-Ray absorption spectroscopies characterize local order around the observed nuclei. With the recent progresses of 71Ga solid state NMR, it has become possible to take advantage of the complementary information that can be obtained using these two methods. This opens the possibility of a more thorough description of the first coordination shells of Ga in oxide. We present and discuss the example of an Al-, Ga-based catalyst. PMID- 10811436 TI - Advances in studies of sociality in nocturnal prosimians. PMID- 10811437 TI - Spatial patterning in nocturnal prosimians: a review of methods and relevance to studies of sociality. AB - Patterns of home range overlap between individuals are key parameters used in discussions of sociality in nocturnal prosimians. Despite the importance of space use variables in defining social structure in nocturnal prosimians, researchers have yet to reach a consensus concerning the most reliable techniques for measuring patterns of home range use. In this paper, we review the methods used in 27 studies of nocturnal prosimian ranging behavior published since 1977. We discuss the usefulness and limitations of the various methods of data collection (radio tracking, trap mark, and identification of sleeping site locations) and data analysis (minimum convex polygon method, minimum concave polygon method, and quadrat analysis) used in these studies. We conclude that the most effective method for gathering data on individual movements and social interactions is direct observation of individual radio tagged animals during all-night follows. In those cases where radio tracking and/or all-night follows are not possible, trap mark techniques can be used, although they tend to greatly underestimate home range size. We recommend that data collected on nocturnal prosimian ranging behavior be analyzed using the minimum convex polygon method, quadrat analysis, and, perhaps, one other of the more mathematically sophisticated techniques popular in studies of non-primate mammals. Finally, we urge researchers to employ standardized methods of data collection and data analysis in future studies of range use in nocturnal prosimians. Without standardization of methods, quantitative comparisons of the findings from different studies are biologically meaningless and prevent cross-species comparisons of space use and its relation to sociality. PMID- 10811438 TI - Sociality in the gray mouse lemur (Microcebus murinus) in northwestern Madagascar. AB - The mating system and social organization of the gray mouse lemur (Microcebus murinus) was investigated in two three-month field studies (covering a period before, during, and after the first mating season) in Ampijoroa, northwestern Madagascar. The spatial and temporal distribution of the sexes within a population was studied using mark/recapture techniques and radiotelemetry to assess possible contest or scramble competition between the males. Sociality was inferred from the occurrence and probability of nocturnal social encounters, the temporal stability of daily sleeping groups, and nocturnal ranging patterns of co sleepers. Males and females were evenly distributed in the study area within a network of highly overlapping home ranges. No indications were found for the spatial monopolization of the females by certain dominant males. Males and females had spatial access to several potential mates; the mating system is therefore characterized as a multi-male/multi-female system. Male home range sizes increased during the first mating season, which was interpreted as an indicator for scramble competition between the males. Competitive mate searching, sperm competition, and temporary mate guarding as well as female mate choice are suggested as the most probable reproductive strategies. Over the course of the study the animals lived continually within the study area, and most females formed stable individualized sleeping associations. Females that slept together shared a higher percentage of their home range than did females that slept at different sites. It is suggested that this network of social relationships should be described as a dispersed but individualized neighborhood. PMID- 10811439 TI - A preliminary study of spatial distribution and mating system of pygmy mouse lemurs (Microcebus cf. myoxinus). AB - According to current hypotheses on the evolution of life history traits and social systems of Malagasy lemurs, nocturnality is associated with a solitary lifestyle and a polygynous or promiscuous mating system. Recent studies, however, have indicated that this may not be true of all lemurs. The goal of this study was to investigate the sociality and the mating system of pygmy mouse lemurs (Microcebus cf myoxinus), which are the smallest known primates, and which retain characteristics of the most primitive primates. I compared my findings with data on the sympatric Microcebus murinus and Cheirogaleus medius. Observational, morphometric and spatial distribution data were obtained by a radiotracking study in 1994, and from a capture-recapture study conducted during 1995/96. Pygmy mouse lemurs usually slept alone in a tangle of vegetation. During the mating season, sleeping sites of males were distributed over a much broader area than were female sites, indicating that male home ranges are larger than those of females. The home ranges of males overlapped during the mating season, and males occasionally roamed over long distances during a single night. Pygmy mouse lemurs forage primarily alone. Analysis of estrus stages indicate that female cycles are unsynchronized during the mating season. There was a lack of sexual dimorphism in body size but not in body mass. Males were heavier than females during the reproductive season but lighter than females the rest of the year. Testes of males varied in size seasonally and enlarged significantly during the mating season. The presence of a vaginal sperm plug in a female indicated the importance of preventing additional matings in this species. Thus pygmy mouse lemurs follow the predictions derived from sexual selection theory for multi-male mating systems with promiscuous matings and male sperm competition. PMID- 10811440 TI - Spatial distribution and population composition of the brown mouse lemur (Microcebus rufus) in Ranomafana National Park, Madagascar, and its implications for social organization. AB - Through a 16-mo mark-recapture trap study, I examined aspects of spatial distribution and population composition in the brown mouse lemur, Microcebus rufus, a 42 g nocturnal strepsirhine. The study took place in the rainforest of Ranomafana National Park in southeastern Madagascar. Sherman live traps were set monthly for a variable number of nights in a quasi-grid 50 m apart. Captured individuals were marked for future identification and released at site of capture. More males than females were captured overall (102 versus 72) and at 83% of trap sites. Trap sex ratio fluctuated significantly over the course of the study. It was particularly male-biased between June and August (3.9:1), when more previously uncaptured males than females (14 versus 6) entered the trap population. Some of these males remained in the trap population. Although the average number of individuals captured was not significantly different between the first four and last four months of the study, the composition of the population changed. The female population, however, changed less: 28.9% of all females captured in the first four months of the study were recaptured in the last four months, compared to 9.7% of males. It is suggested that the pattern of appearance of new individuals and disappearance of others, both predominantly male, may indicate migratory activity. Furthermore, an average of eight individuals were captured at each trap site (approximately 70% of traps captured more than five), suggesting a high degree of spatial overlap. The average number of male and female individuals captured in each trap (5.5 males versus 2.5 females), the average number of trap sites at which males and females were captured (3.6 versus 2.4), and the average number of captures for males and females (9.8 versus 5.7) all differed significantly between the sexes. PMID- 10811441 TI - Preliminary observations on sexual behavior and the mating system in free-ranging lesser galagos. (Galago moholi). AB - Sexual and associated patterns of behavior of lesser galagos (Galago moholi) were recorded during an 18-month study conducted at the Nylsvley Nature Reserve in South Africa. Animals were trapped and fitted with radio transmitter belts in order to monitor nocturnal activities during twice-yearly mating seasons. Most copulations occurred during the last week in May, while a subsidiary (post partum) mating season occurred in late September-early October. Females came into estrus sequentially during the May season. Adult males exhibited increase in body weight and testes volume during the mating season, changes which were most pronounced among the larger males (> 226 g). Larger males also had the greatest mating success, initiating 88% of observed copulations. Sixty-seven percent of matings involved more than one male copulating with the same female during her estrus, which lasted 1-3 days. Mounts were prolonged (range 2-53 min, mean 9.0 min) and males copulated repeatedly (2-5 times) with the same partner during a single night. These observations of sexual behavior and of large relative testes size in free-ranging lesser galagos are consistent with the occurrence of a dispersed mating system involving sperm competition in this nocturnal prosimian species. PMID- 10811442 TI - Sociality in the spectral tarsier, Tarsius spectrum. AB - Recent studies indicate that many of the nocturnal prosimian primates are gregarious rather than solitary. This paper shows that the spectral tarsier is gregarious during its nightly activity period as well as in its sleeping tree. Using mist nets and radiotelemetry, focal follows were conducted on six groups at Tangkoko Nature Reserve in Sulawesi, Indonesia. During 442 focal follows, 1072 encounters between a focal adult group member and another adult were observed. The number of encounters ranged from as few as 0 to as many as 18 encounters per night. Intragroup encounters lasted from less than 1 min to as long as 3 hr 12 min. Nearly one-half of all social behavior occurred between adult females and males. There were also substantial rates of social interaction between the two adult females in one group, and between sub-adults of the opposite sex in neighboring groups. PMID- 10811443 TI - A role for P-glycoprotein in regulating cell death. AB - P-glycoprotein (P-gp) is an energy dependent drug pump responsible for multidrug resistance (MDR) in human cancers. While it is irrefutable that P-gp can efflux xenobiotics out of cells, the biological function of P-gp in multicellular organisms has yet to be firmly established. The question of what, if anything, P gp does when not effluxing drugs has been raised by recent reports indicating that P-gp may regulate apoptosis, chloride channel activity, cholesterol metabolism and immune cell function. There is now a lively debate regarding the possible role of P-gp in regulating cell differentiation, proliferation and survival. PMID- 10811444 TI - Defective megakaryocytic development in myelodysplastic syndromes. AB - Megakaryocytic proliferation and differentiation is typically abnormal in patients with myelodysplastic syndromes (MDS). The underlying mechanisms for this finding are not known, but may involve defects at the level of the thrombopoietin receptor (c-mpl) or post-receptor signaling pathways in megakaryocyte progenitor cells. Premature apoptosis of the bone marrow cells and inhibitory effects of cytokines such as tumor necrosis factor alpha have been implicated as contributing to altered megakaryopoiesis in MDS, but their significance remains unclear. The availability of thrombopoietin (TPO) has facilitated more detailed analysis of megakaryocytic biology using several experimental in-vitro systems. However numerous studies have shown that the developmental abnormalities of MDS megakaryocytes could not be corrected by TPO. Increasing investigations are being extended to the evaluation of signal transduction pathways of c-mpl both in cell lines and human hematopoietic cells in order to identify the molecular mechanisms responsible for the defective megakaryocytic development in MDS. PMID- 10811445 TI - Role of interferon-alpha and clonally expanded T cells in the immunotherapy of chronic myelogenous leukemia. AB - Twenty five percent of patients in the chronic phase of chronic myelogenous leukemia (CML) are treated with interferon-alpha (IFN-alpha) to induce a cytogenic remission. In addition to its direct effects on leukemic cells, IFN alpha has been shown to induce immunologic alterations, including upregulation of the expression of major histocompatibility (MHC) antigens in antigen-presenting cells (APCs), as well as augmentation of the activity of the lymphocytes against tumor cells. However, there has been little direct evidence supporting a causal interaction between cellular immunoreactivity and clinical responsiveness to IFN alpha. We have shown that one approach to elucidate the immunological mechanisms by which IFN-alpha exerts its anti-CML activity is by analyzing therapy-induced modulation in T-cell receptor (TCR) Vbeta chain usage, using the reverse transcription-polymerase chain reaction (RT-PCR) followed by single-strand conformation (SSCP) analysis. This method is particularly attractive, since it provides an index of antigen-specific T cell expansion, but does not require the extraction and purification of the antigens involved in the T-cell response. T cell clones that express the Vbeta 10, 12, and 14 families predominate in the peripheral blood (PB) of CML patients. The enhanced expression of the Vbeta 9 and 20 families has been detected in IFN-alpha responsive patients but not patients who are poorly responsive to this agent. This suggests that expansion of T cells expressing these TCR Vbeta gene families may serve as a prognostic factors of the clinical responsiveness of CML patients to IFN-alpha. In addition, since T cell clones that express certain Vbeta families may react with a discrete set of antigenic peptides presented on the surface of malignant cells, a better understanding of the immunobiology of T cells in CML may allow for the design of increasing efficacious immune therapy for this disease. PMID- 10811446 TI - Interleukin-8 and other agonists of Gi2 proteins: autocrine paracrine growth factors for human hematopoietic progenitors acting in synergy with colony stimulating factors. AB - We have reviewed the current knowledge on CXC chemokine interleukin-8 (IL-8) and human hematopoiesis, and more generally on agonists of heterotrimeric Gi2 proteins as regulators of human hematopoiesis. It appears that low doses of IL-8, a Gi2-agonist produced in an autocrine fashion by normal hematopoietic progenitors, mature blood cells and leukemic cells, promotes cell survival or/and proliferation in response to hematopoietic cytokines. More importantly, inactivation of the IL-8/Gi2 pathways inhibits CD34+ cell proliferation and colony formation. Similar positive effects on hematopoiesis of other, physiological or pathological, agonists of Gi2 proteins are discussed, as well as the molecular pathways involved and the consequences of activation of other G proteins (Gq, G16) by IL-8 and other Gi2-agonists. PMID- 10811447 TI - The janus kinase inhibitor, Jab/SOCS-1, is an interferon-gamma inducible gene and determines the sensitivity to interferons. AB - The Janus family of protein tyrosine kinases (JAKs) and STAT transcription factors regulate cellular processes involved in cell growth, differentiation, and transformation through their association with cytokine receptors. The CIS family of proteins (also referred as the SOCS or SSI family) has been implicated in the regulation of signal transduction by a variety of cytokines. Among them, we have shown that JAB/SOCS-1 is strongly induced by interferon-gamma and forced expression of JAB/SOCS-1I conferred cells interferon resistance. This resistance was caused by inhibition of JAK1 and JAK2 activation in response to IFNgamma. Moreover, recent detailed analysis of JAB/SOCS-1 knockout mice revealed that JAB/SOCS-1 is indeed a "negative feedback regulator" that determine the sensitivity of cells to IFNgamma. Using in vitro mutagensis, we defined a functional structure of JAB/SOCS-1 and proposed a mechanism for how JAB inhibits JAK kinase activity. PMID- 10811448 TI - Alternative pathways of cell death to circumvent pleiotropic resistance in myeloma cells: role of cytotoxic T-lymphocytes. AB - Pleiotropic resistance to treatment remains one of the major reasons for therapeutic failures in patients with multiple myeloma. Myeloma cells are frequently resistant to physiological inducers of cell death prior to chemotherapy. Moreover, in the course of treatment cells acquire a multidrug resistant (MDR) phenotype, making eradication of the tumor even more difficult. A necessary prerequisite for circumventing complex pleiotropic resistance is therefore defining the signaling pathways that execute death in myeloma cells. This review discusses evidence that cytokine-expressing autologous tumor cell vaccine may be an efficient tool for elimination of both intrinsically resistant myeloma cells as well as cells with acquired MDR in murine models. The vaccine was similarly potent against wild type cells that were resistant to several death receptor ligands, and their isogenic sublines selected for P-glycoprotein mediated MDR. The anti-myeloma effect of the vaccine was mediated by granzyme B/perforin-secreting cytotoxic T-lymphocytes. This is an example of therapeutic strategy directed at utilizing death pathways that are preserved in pleiotropically resistant tumor cells. PMID- 10811449 TI - Integrin-mediated drug resistance in multiple myeloma. AB - Drug resistance remains a major obstacle to the treatment of many hematopoietic malignancies such as multiple myeloma. Although much research has been focused on acquired resistance phenotypes, we believe that de novo drug resistance mechanisms may be an important component in protecting cells from initial drug exposure. It is now realized that many of the biological processes associated with this disease, including cell survival, may come as a result of the direct interactions of malignant plasma cells with the bone marrow microenvironment. This review examines the role of cell adhesion to one bone marrow component, fibronectin (FN), and the impact it may have on response to cytotoxic drugs. We discuss the influence of the integrin VLA-4 (alpha4beta1) on cell adhesion mediated drug resistance (CAM-DR) as well as the effects of chronic drug exposure on integrin function. Data presented here demonstrates that drug selection can make a non-adherent cell line adherent to FN through inside-out integrin activation and consequently cause a decrease in sensitivity to drug. We also speculate on the possible mediators of this intrinsic mechanism of drug resistance which may, along with the integrins themselves, become promising therapeutic targets in cancer treatment. PMID- 10811450 TI - Blast crisis of Ph-positive chronic myeloid leukemia with isochromosome 17q: report of 12 cases and review of the literature. AB - Isochromosome 17q [i(17q)] is frequently observed in the blast crisis (BC) of chronic myelogenous leukemia (CML). It has been suggested that this chromosome abnormality is associated with special hematological characteristics of the BC, but the information on this subject is scarce. The clinical, hematological and cytogenetic features of patients with i(17q) were analyzed in a series of 121 patients with BC of Ph-positive CML. Twelve patients (10%) displayed an i(17q), representing the third commonest cytogenetic abnormality, after trisomy 8 and Ph chromosome duplication. In seven of the 12 patients the BC was preceded by an accelerated phase, and 10 had more than 10% blood basophils at BC diagnosis. The blast cells had a myeloid phenotype in the 12 patients. Five patients exhibited cytogenetic abnormalities in addition to i(17q), with trisomy 8 and duplication of the Ph chromosome being the alterations most frequently observed. Median survival of patients with i(17q) was 22 weeks, which was not significantly different from the survival of patients with myeloid BC in the overall series. These results are similar to the findings in 181 patients with i(17q) from 12 series of the literature, and confirm the special hematologic profile of BC of CML with this cytogenetic abnormality. PMID- 10811451 TI - Phase I/II trial of multiple dose 131Iodine-MAb LL2 (CD22) in patients with recurrent non-Hodgkin's lymphoma. AB - The purpose of this to evaluate in a phase I/II study the efficacy and toxicity of a multi-dose administration of 131I labeled CD22 monoclonal antibody (131I-MAb LL2) in escalating dose cohorts administered to relapsed non-Hodgkin's lymphoma (NHL) patients. Twenty-one patients with relapsed NHL received one of four dose levels of 131-MAb-LL2 administered in a twice weekly pattern. Starting with dose level 2, the patients also received 20 mg of unlabeled LL2 prior to each radiolabeled dose administered. Previously stored autologous peripheral blood progenitors were reinfused in case of prolonged cytopenias. Patients could repeat the same treatment if they had stable disease or a response to the first therapy at 8 weeks, and had not received their peripheral blood progenitors with the first cycle. Combining all of the dose cohorts, there were 5 complete responses or complete responses (undetermined) and 2 partial responses for a total response rate of 7/21 (33%). There was no dose response effect with responses documented at all dose levels. Expected toxicities were hematopoietic, requiring stem cell re-infusion in 5 patients. Other toxicities included hypothyroidism in 3 patients, and human anti-mouse antibody formation (HAMA) in 4 patients. In conclusion, 131I-MAb-LL2, when administered in a multi-dose fashion with 20 mg unlabeled antibody pre-dosing, resulted in a response rate of 33% in heavily pre treated NHL patients. Non-hematologic toxicities were mild and few in number. Further evaluation of this treatment is warranted and further dose escalation will be possible. PMID- 10811452 TI - Autologous peripheral blood stem cell transplantation for adults with B-lineage acute lymphoblastic leukemia: a pilot study. AB - We conducted a pilot study on autologous peripheral blood stem cell transplantation (PBSCT) for 11 adults with B-lineage acute lymphoblastic leukemia (ALL) in first complete remission (CR) or even in those with more advanced stages. All patients achieved CR by induction therapy, of whom 10 were treated with anthracycline, vincristine and prednisolone-based regimens. After consolidation therapy, all patients except one received high-dose cytarabine followed by granulocyte colony-stimulating factor (G-CSF) administration to collect PBSCs. Ten patients received busulfan 4 mg/kg for 4 days, etoposide 20 mg/kg for 3 days and ranimustine 200 mg/m2 for 2 days as a conditioning regimen. One received a regimen consisting of etoposide, cyclophosphamide and total body irradiation. From day 1, G-CSF was given intravenously, and no additional chemotherapy was administered. At the median follow-up time of 30.8 months, four of six patients with standard-risk B-lineage ALL survived within the range of 19.7 to 85.4 months without relapse. In contrast, only one of five with high-risk B-lineage ALL survived for 36.3 months without relapse. Autologous PBSCT as post remission therapy may prolong CR in adults with standard-risk B-lineage ALL. PMID- 10811453 TI - Plasma erythropoietin in essential thrombocythaemia: at diagnosis and in response to myelosuppressive treatment. AB - Sixty-five patients with essential thrombocythaemia (ET) on different treatment regimens were studied with regard to EDTA-plasma erythropoietin (EPO) concentrations. In accordance with other studies we found that close to 50% of the untreated ET patients had subnormal (<3.7 IU/L) plasma EPO. The mean plasma EPO concentration for untreated ET patients was significantly lower compared to patients treated with hydroxyurea (HU), radiophosphorous, alpha-interferon or combinations of myelosuppressive agents. This was also true after correction for differences in haemoglobin concentrations had been introduced. An increase in plasma EPO was recorded in all 20 ET patients in whom plasma EPO was registered before and after initiation of myelosuppressive therapy. At the time of diagnosis plasma EPO concentration was available in 31 of the ET patients. In 13 of them the plasma EPO was subnormal whereas the EPO concentrations were > or =3.7 IU/L in the remaining 18 subjects. It was demonstrated that the time to initiation of myelosuppressive treatment was significantly shorter for the former group of patients; they also had more vascular events (11 out of 13) than the group of patients with plasma EPO concentrations > or =3.7 IU/L (9 out of 18). It therefore appears that a subnormal plasma EPO in newly diagnosed ET might be a risk factor for the development of vascular complications. PMID- 10811454 TI - B-cell lymphoma-associated hemophagocytic syndrome. AB - B-cell lymphoma-associated hemophagocytic syndrome (B-LAHS) is extremely rare in Western countries but has recently been increasingly reported in Asian countries, especially Japan. Here, we reviewed 25 previously reported Japanese cases of B LAHS and summarized its clinicopathologic features and therapeutic outcome. The median age of onset was 63 years old with initial presentation of fever, hepatomegaly, and splenomegaly without associated lymphadenopathy. Laboratory findings showed increased levels of lactate dehydrogenase, C-reactive protein, ferritin and soluble interleukin-2 receptor. Histopathologically, hemophagocytosis was often seen in the bone marrow and spleen. Various percentages of lymphoma cells were seen in the bone marrow, positive for CD19, CD20 and surface immunoglobulin. and some were also positive for CD5. Cytogenetic analysis showed a complex structural abnormality including chromosome 14q32, 19q13 and deletion of the terminal part of 8p21. Some patients had histological features of intravascular lymphomatosis (IVL). The prognosis was poor with a median survival period of 9 months. We treated five patients using autologous peripheral blood stem cell transplantation (PBSCT), and four are still in complete remission nine to 24 months after PBSCT, suggesting that high-dose chemotherapy followed by PBSCT might improve the survival rate. PMID- 10811455 TI - The association of thyrotoxicosis and chronic lymphoproliferative disorders. AB - We diagnosed thyrotoxicosis (Tx) and chronic lymphoproliferative disease (LPD) in nine cases; seven had chronic lymphocytic leukemia (CLL), one had hairy cell leukemia (HCL) and one had non-Hodgkin's lymphoma (NHL). In a literature review, we found only one reported case with Tx and CLL and documentation of an higher incidence of Tx in the relatives of CLL patients. We report these nine cases here and discuss this interesting association. PMID- 10811456 TI - Relapse of leukemia and lymphoma after marrow transplant: a review of cases with extramedullary relapse. AB - We review our cases of leukemia and lymphoma relapse after allogeneic marrow transplant and describe here a series of 10 patients with extramedullary (EM) relapse. In the 13 relapses in acute myeloid leukemia, 5 cases had EM involvement. There were 3 EM involvement out of 13 acute lymphoblastic leukemia relapses, one EM disease in 11 chronic myeloid leukemia relapses and one case of lymphoma with EM relapse. A common observation is that in some of these cases, EM relapse occurred in the presence of continuous marrow remission, In those cases with both marrow and EM involvement marrow remission could often be achieved and maintained temporarily while EM disease progressed despite chemotherapy or immunotherapeutic measures such as immunosuppressant withdrawal and donor lymphocyte infusion. Survival in partial remission after relapse could be prolonged in some cases but eventual death from progressive disease was often the case. PMID- 10811457 TI - Stromal cell CD9 and the differentiation of hematopoietic stem/progenitor cells. AB - CD9 belongs to the tetraspan family of proteins that facilitates the regulation of cell proliferation, motility, and adhesion. In mouse hematopoietic organs, CD9 is expressed by myeloid and stromal cells. Although the precise mechanisms are not clear, antibody ligation of CD9 on stromal cells regulates the adhesion between stromal cells and hematopoietic stem cells, the production of myeloid cells in long term bone marrow cultures and the differentiation of hematopoietic stem cells. A 100 kD protein co-precipitated with CD9 is distinct from several previously reported CD9-associated molecules with respect to size and distribution. Identification and analysis of this interesting protein may clarify the molecular mechanisms through which CD9 bearing stromal cells control the differentiation of hematopoietic stem cells and/or allow them to maintain their vital self-renewal capacity. PMID- 10811458 TI - The effects of arsenic trioxide (As2O3) on human megakaryocytic leukemia cell lines. With a comparison of its effects on other cell lineages. AB - Arsenic trioxide (As2O3) has been demonstrated to be effective for the treatment of acute promyelocytic leukemia (APL) and to inhibit proliferation and produce apoptosis in the APL cell line NB4. The effect of this newly utilized chemotherapeutic agent on other lineages is currently under study to evaluate its efficacy for the treatment of other human malignancies and myeloproliferative syndromes. A recent study described the effects of As2O3 upon viability, proliferation, and induction of apoptosis in four different megakaryocytic leukemia cell lines. At pharmacological concentrations (0.5-2 microM) As2O3 selectively inhibits growth and causes apoptosis in the megakaryocytic leukemia cell lines HEL, Meg-01, UT7 and M07e. Pertinently, these concentrations of As2O3 resulted in identical changes in the characteristics of the APL cell line NB4, suggesting that As2O3 could produce its effects in both cellular lineages via a common mechanism of action. Various mechanisms have been proposed for the As2O3 induced changes in NB4 (including modulation of promyelocytic leukemia proteins (PML) and Bcl-2, modification of the glutathione redox system, caspase activation, and cell cycle arrest) and are currently under investigation in the megakaryocytic leukemia cell lines. Recent preliminary results indicate that As2O3 downregulates Bcl-2 expression and induces cell cycle arrest in megakaryocytic cell lines. The use of As2O3 for the treatment of malignant megakaryocytic disorders also has been considered. The in vitro effects of As2O3 on a chronic megakaryocytic proliferative disorder. i.e., Essential Thrombocythemia (ET), have been analyzed and megakaryocyte progenitors have shown an unexpectedly higher resistance to As2O3, in comparison to normal megakaryocyte colony-forming cells. The effects of As2O3 on ET and other megakaryocytic disorders need to be fully examined, in order to determine the clinical efficacy of As2O3 in the treatment of syndromes affecting the megakaryocytic lineage. PMID- 10811459 TI - Fibre content and cellularity of the bone marrow of the iliac crest, vertebral column and sternum in chronic myeloproliferative disorders. AB - Heterogeneous content of fibres and haematopoesis within the bone marrow may affect diagnosis and staging in chronic myeloproliferative disorders (CMPDs). To evaluate their distribution, we conducted a post mortem histomorphometric study of 22 patients with CMPD in chronic phases. In bone marrow specimens from the anterior and posterior iliac crest (right and left of each), the sternum, the 7th thoracic and the 3rd lumbar vertebra, the argyrophil fibres were counted using the line intersection method and the cellular and fatty bone marrow using the point count method. Statistical analysis was performed by direct comparison of the sites. The distribution of fibres was almost homogeneous in the patients with low fibre content, revealing a random diversity in more advanced stages of marrow fibrosis. 1/22 patient had no fibre increase in one specimen of the iliac crest and overt myelofibrosis in the other sites. 1/22 patient had myelofibrosis in two sites of the iliac crest and no fibre increase in vertebral column and sternum. The bone marrow cellularity was almost homogeneously increased in all patients. Myelofibrosis proved to be a generalised process with heterogeneous grades of severity in different regions of the bone marrow in CMPDs. No topographical bias was found. In contrast to the homogeneous increase of the bone marrow cellularity the topographical heterogeneity of the fibre content may limit the representativity of single bone marrow biopsies in patients with CMPDs. PMID- 10811460 TI - BCR-ABL directed immunotherapy: a virtual reality? AB - Nearly ten years of research on the feasibility of specific immunotherapy targeting the junctional regions of BCR-ABL has considerably increased our knowledge of which MHC alleles might present BCR-ABL peptides, yet has failed to provide us with definite proof of appropriate processing of the hybrid oncoprotein into such antigenic peptides. This paper intends to provide an overview of the current state of affairs as well as to delineate limitations and future directions of this line of research. PMID- 10811461 TI - Progressive multifocal leukoencephalopathy with concurrent Richter's syndrome. AB - Progressive multifocal leukoencephalopathy (PML) is a demyelinating infectious disease caused by the JC virus. It was originally described in patients with chronic lymphocytic leukemia (CLL). Richter's syndrome, or transformation to large cell Lymphoma, occurs in approximately 3% of patients with CLL, and carries a poor prognosis. We report a patient with documented PML and concurrent Richter's transformation outside the central nervous system. Before establishing a definitive diagnosis of PML, radiation therapy to the presumed lymphomatous brain lesion had been considered, raising the issue of whether stereotactic brain biopsy should be considered in every patient in a similar situation. Although this is likely a rare occurrence, patients with Richter's transformation documented at an extra-neural site and a brain lesion may benefit from the establishment of an infectious diagnosis which would influence therapy. PMID- 10811462 TI - Successful treatment of all-trans retinoic acid resistant and chemotherapy naive acute promyelocytic patients with arsenic trioxide--two case reports. AB - Arsenic trioxide(As2O3) has proved highly effective in treating both refractory or primary cases of acute promyelocytic leukemia (APL). The role of arsenic trioxide in APL treatment has been confirmed by study groups in China and in the USA. However, what is the role of As2O3 in treating APL? Should it be used as first line therapy, or should it be used as a second line drug. This still remains to be defined. Here, we report two cases of APL, who were treated successfully with As2O3 when they relapsed. Initially, both received all-trans retinoic acid (ATRA) for primary remission induction therapy, and obtained a complete remission. For ethical or personal reasons, they did not receive chemotherapy as consolidation therapy and when they relapsed at 23 months and 12 months later respectively, they both received As2O3 therapy after being resistant to ATRA treatment. Two courses of As2O3 were given and both reached complete remission. There were very few adverse reactions to the drug, only mild abdominal cramps, mild fluid retention, and transient elevation of transaminases. They both had rather good quality of life throughout the treatment and both remain in remission for 32 months and 10 months since therapy, respectively. PMID- 10811463 TI - Retinoic acid syndrome induced by arsenic trioxide in treating recurrent all trans retinoic acid resistant acute promyelocytic leukemia. AB - Arsenic Trioxide (As2O3) is an effective agent for treating acute promyelocytic leukemia achieving a complete remission rate of about 60% to 90%. It is similar to all-trans retinoic acid (ATRA) when treating acute promyelocytic leukemia (APL), because both agents have limited side effects compared to conventional chemotherapy, although the treatment period is more prolonged. During treatment, both agents may induce leukocytosis, and in patients taking ATRA, leukocytosis appears to be related to the development of retinoic acid syndrome (RAS). We report here a case of APL treated with ATRA in combination with chemotherapy 3 years earlier. During treatment, an episode of RAS with fever, edema, pericardiac effusion etc. was encountered. Recently, she had a relapse of leukemia, and As2O3 therapy was used. Leukocytosis developed again, and symptoms of fever, skin rash, edema resembling a RAS also developed, which was quickly relieved by steroid administration in a manner resembling response to RAS. PMID- 10811464 TI - CD-30 positive peripheral T-cell lymphoma of the Waldeyer's ring. AB - We describe a patient with peripheral T cell lymphoma of the Waldeyer's ring that ran a highly aggressive course. This is followed by a discussion on the differential diagnoses of nasal T/NK cell lymphoma and Ki-1 ALCL, based on clinical and pathological features. PMID- 10811465 TI - Successful treatment of primary adrenal non-Hodgkin's lymphoma associated with adrenal insufficiency. AB - We report a case of primary adrenal NHL associated with adrenal insufficiency which was successfully treated with steroid replacement and chemotherapy. A 69 year-old woman hospitalized with fatigue and weight loss developed shock and recovered with steroid therapy. Adrenal insufficiency was confirmed by an elevated plasma adrenocorticotropic hormone level and low cortisol. Computed tomography revealed large bilateral adrenal masses. Needle biopsy showed a diffuse, mixed B cell lymphoma. CHOP therapy accompanied by steroid replacement was begun, and she achieved a complete remission after 4 cycles. She received additional 4 cycles of chemotherapy. Although adrenal insufficiency was irreversible, she has continued in complete remission for 50 months at this reporting. PMID- 10811466 TI - An unusual case of chylothorax complicating non-Hodgkin's lymphoma. AB - A 64 year old man with non-Hodgkin's lymphoma developed extremely troublesome chylous pleural effusions following effective chemotherapy. With the pre operative use of olive oil, a diffuse leakage of lymph was seen at thoracotomy. Oversewing was performed eventually resulting in an excellent outcome. PMID- 10811467 TI - A new cholesterol derivative suitable for transfecting certain type of cells in the presence of 10% serum. AB - We developed a new cationic lipid suitable for use as a DNA carrier in the presence of 10% sera. The novel compound (abbreviated as Arg-Chol) contains cholesterol and a dipeptide consisting of glycine and sterically protected arginine. The efficiency of reporter gene transfection using liposomes based on this new reagent was compared with that of liposomes made with other cationic derivatives of cholesterol. Lipoplexes formulated with the newly synthesized lipid mediate in vitro transfection of B16(F10) murine melanoma cells in the presence of 10% sera more efficiently than in other cell lines and compared with other cholesterol derivatives studied. PMID- 10811468 TI - The potential of acetaminophen as a prodrug in gene-directed enzyme prodrug therapy. AB - Acetaminophen is oxidized by human CYP1A2 to the cytotoxic metabolite N acetylbenzoquinoneimine (NABQI). Incubation of cells transfected with human CYP1A2 (H1A2 MZ cells) with 4-20 mM acetaminophen for 6 hours at 37 degrees C caused extensive cytotoxicity (cell viability <10%). In contrast, nontransfected V79 MZ cells were unaffected (viability >95%). By mixing H1A2 MZ cells with V79 MZ cells in various proportions and incubating with 4 mM acetaminophen, it was shown that the NABQI released from H1A2 MZ cells also caused cytotoxicity of bystander cells. Thus, in a mixture containing 5% H1A2 MZ cells, exposure to 4 mM acetaminophen for 6 hours resulted in complete cell killing by 24 hours. A similar bystander effect was found by incubating the same proportion of CYP1A2 containing cells with ovarian tumor-derived SK-OV-3 cells or colon tumor-derived HCT116 cells. However, breast tumor-derived MDA-MB-361 cells displayed resistance to the cytotoxic effect of NABQI, and it was necessary to increase the proportion of H1A2 MZ cells to 50% to achieve complete cell killing. In conclusion, the use of acetaminophen as prodrug and CYP1A2 as an activating enzyme is a promising combination for gene-directed enzyme prodrug therapy. PMID- 10811469 TI - Pharmacologic suppression of target cell recognition by engineered T cells expressing chimeric T-cell receptors. AB - Adoptive therapy with autologous T cells expressing chimeric T-cell receptors (chTCRs) is of potential interest for the treatment of malignancy. To limit possible T-cell-mediated damage to normal tissues that weakly express the targeted tumor antigen (Ag), we have tested a strategy for the suppression of target cell recognition by engineered T cells. Jurkat T cells were transduced with an anti-hapten chTCR tinder the control of a tetracycline-suppressible promoter and were shown to respond to Ag-positive (hapten-coated) but not to Ag negative target cells. The engineered T cells were then reacted with hapten coated target cells at different effector to target cell ratios before and after exposure to tetracycline. When the engineered T cells were treated with tetracycline, expression of the chTCR was greatly decreased and recognition of the hapten-coated target cells was completely suppressed. Tetracycline-mediated suppression of target cell recognition by engineered T cells may be a useful strategy to limit the toxicity of the approach to cancer gene therapy. PMID- 10811470 TI - Cellular and humoral immune responses to adenovirus and p53 protein antigens in patients following intratumoral injection of an adenovirus vector expressing wild type. P53 (Ad-p53). AB - The immune responses of 10 patients with advanced non-small cell lung cancer receiving monthly intratumoral injections of a recombinant adenovirus containing human wild-type p53 (Ad-p53) to adenovirus and transgene antigens were studied. The predominate cellular and humoral immune responses as measured by lymphocyte proliferation and neutralizing antibody (Ab) formation were to adenovirus serotype 5 vector antigens, with increased responses in posttreatment samples. Consistent alterations in posttreatment cellular and humoral immune responses to p53 epitopes were not observed, and cytotoxic Abs to human lung cancer cells were not generated. Patients in this study had evidence of an antitumoral effect of this treatment with prolonged tumor stability or regression; however, neither Abs to p53 protein nor increased lymphocyte proliferative responses to wild-type or mutant p53 peptides have been consistently detected. PMID- 10811471 TI - Synergistic effects of adenovirus expressing wild-type p53 on chemosensitivity of non-small cell lung cancer cells. AB - The infection of recombinant adenovirus expressing wild-type p53 (Ad-p53) to lung cancer cells that harbor mutant p53 genes improves their response to cis diamminedichloroplatinum(II). In this study, we tested whether this improvement in response is also seen in wild-type p53 (wt-p53)-containing cancer cells and whether this phenomenon is universal with other commonly used chemotherapeutic agents, including etoposide, 7-ethyl-10-hydrocycamptothecin, paclitaxel, and docetaxel. Using a panel of 7 non-small cell lung cancer cell lines with wild type (2) or abnormal (2 null, 3 point-mutated) p53, we examined in vitro cytotoxicity using a tetrazolium-based colorimetric assay (3-(4,5-diethylthiazoyl 2-yl)-2,5-diphenyltetrazolium bromide assay) and analyzed the combined effects of Ad-p53 and chemotherapeutic agents using the isobologram method. Ad-p53 and DNA damaging agents (cis-diamminedichloroplatinum(II), etoposide, and 7-ethyl-10 hydrocycamptothecin) showed synergistic effects in six of seven cell lines but additive effects against a p53-mutated cell line. In contrast, Ad-p53 showed additive effects with the antitubulin agents (paclitaxel and docetaxel) in all four of the cell lines tested. Furthermore, we examined this synergistic interaction between Ad-p53 and DNA-damaging agents by flow cytometric analysis and DNA fragmentation analysis. Both analyses revealed that a sublethal dose of Ad-p53 augmented the apoptotic response induced by DNA-damaging agents in six of seven cell lines. Our results suggest that Ad-p53 may synergistically enhance the chemosensitivity of the majority of non-small cell lung cancers to DNA-damaging agents due to augmentation of apoptosis. PMID- 10811472 TI - Modulation of drug cytotoxicity by reintroduction of wild-type p53 gene (Ad5CMV p53) in human pancreatic cancer. AB - Chemotherapy does not significantly improve prognosis in pancreatic cancer. New therapeutical approaches involving p53 gene replacement appear to be very encouraging due to the key role of p53 in the cell response to DNA damage. Here, we have evaluated the effectiveness of combining wild-type p53 (wt-p53) gene reintroduction (Ad5CMV-p53) and exposure to two genotoxic drugs, gemcitabine and cisplatin, in several human pancreatic cell lines. The efficiency of the combinations was clearly dependent upon timing, as assessed by cell survival determinations. Although wt-p53 transduction before drug treatment induced chemoresistance, p53 transduction in cells treated previously with gemcitabine increased cytotoxicity. Cell cycle profiles showed significant decreases in the percentage of cells in the S phase as a consequence of arrests provoked by the expression of exogenous p53, reducing the number of cells susceptible to the drug. The sensitivity of cells to cisplatin, which has a lower degree of S-phase specificity, was not modified as much by p53 gene replacement. In contrast, the recognition of the previous drug-induced DNA damage by the newly expressed wt-p53 elicited increases in sub-G1 populations, consistent with the annexin determinations and bax/bcl-2 ratios observed. Experiments on subcutaneous pancreatic xenografts corroborated the effectiveness of this approach in vivo. Thus, the combination of p53 transduction and chemotherapy, under a correct schedule of administration, appears to be a very promising therapy for human pancreatic cancer. PMID- 10811473 TI - Avoidance of bone marrow suppression using A-5021 as a nucleoside analog for retrovirus-mediated herpes simplex virus type I thymidine kinase gene therapy. AB - Gene therapy using the herpes simplex virus thymidine kinase (HSV-TK) gene combined with an anti-herpes drug, ganciclovir (GCV), has been applied for human diseases, especially for cancer treatment. However, bone marrow toxicity has been the most consistent adverse effect of GCV treatment in clinical settings. We evaluated the cytotoxic activity of a novel guanosine analog, (1'S,2'R)-9[[1',2' bis(hydroxymethyl)cycloprop-1'-yl]methyl]guanin e (A-5021), against retrovirus mediated HSV-TK gene-transduced human lung cancer cells. The bone marrow toxicity of A-5021 and GCV was studied by colony formation assay in both rodent and human bone marrow specimens. We demonstrated that A-5021 had potent cytotoxic activity equal to that of GCV against the retroviral vector-mediated HSV-TK gene transduced lung cancer cell lines. Further, phosphorylated A-5021 could be transferred to neighboring cells, and this analog killed HSV-TK- neighboring cells, as was the case for GCV. In contrast, A-5021 did not exhibit an inhibitory effect on bone marrow progenitor cells and colony formation (the 50% inhibitory concentration of the colony-forming units in culture = >100 microg/mL in human bone marrow specimens and >66 microg/mL in rodent bone marrow specimens). These results indicate that A-5021 has potent cytotoxic activity as a nucleoside analog for gene therapy using HSV-TK gene, and can be used much more safely than GCV. PMID- 10811475 TI - Use of a herpes thymidine kinase/neomycin phosphotransferase chimeric gene for metabolic suicide gene transfer. AB - Metabolic suicide gene transfer is widely applied for gene therapy of cancer, and retroviral vectors expressing the herpes simplex virus thymidine kinase (HSV-tk) gene are commonly used in clinical trials. Most of these vectors contain positive selectable markers that undoubtedly facilitate the determination of viral titer and the identification of high-titer producer clones. However, the presence of additional transcriptional units may result in reduced expression of the gene of interest. The use of fusion genes expressing bifunctional proteins may help to overcome this problem. We have constructed a retroviral vector carrying the TNFUS69 chimeric gene, which originates from the fusion of the HSV-tk and neomycin phosphotransferase II genes, and evaluated the functional expression of the encoded fusion protein. In vitro, expression of the fusion gene conferred to target cells both resistance to neomycin and selective sensitivity to the antiherpetic drugs ganciclovir and (E)-5-(2-bromovinyl)-2'-deoxyuridine. Cells transduced with the fusion gene, however, showed reduced ability to phosphorylate ganciclovir compared with cells expressing the native HSV-tk. Therefore, although the fusion gene may be used as a constituent of retroviral cassettes for positive and negative selection in vitro, its usefulness for suicide gene transfer applications in vivo may depend upon the possibility of using (E)-5-(2 bromovinyl)-2'-deoxyuridine in a clinical context. PMID- 10811474 TI - Two-drug combinations that increase apoptosis and modulate bak and bcl-X(L) expression in human colon tumor cell lines transduced with herpes simplex virus thymidine kinase. AB - Herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) gene therapy can induce apoptosis in tumor cells that are normally resistant to this type of cell death, although the cellular mechanisms by which this occurs remain to be elucidated. Human colon tumor cell lines expressing HSV-TK were treated with GCV or four other inducers of apoptosis: butyrate, camptothecin (CPT), Taxol (paclitaxel), or 7-hydroxystaurosporine (UCN-01). Over a 2-4 day treatment period with GCV or the other four drugs, protein levels of the apoptosis agonist Bak increased 1.5- to 3-fold, whereas a corresponding decrease in the levels of the apoptosis antagonist, Bcl-X(L), was observed in butyrate-, CPT-, and 7 hydroxystaurosporine (UCN-01)-treated cells. GCV and paclitaxel treatments resulted in increased levels of Bcl-X(L). In two-drug combinations with GCV plus one of the four other drugs, increased tumor cell killing was found with GCV plus UCN-01 or with some GCV/butyrate combinations; the other two tested combinations were largely antagonistic. The GCV/UCN-01 and GCV/butyrate combinations resulted in increased Bak and decreased Bcl-X(L) protein levels, while the GCV/CPT and GCV/paclitaxel combinations resulted in increased levels of both proteins. The results highlight the potential for new combination therapies of HSV-TK/GCV and chemotherapeutic drugs that result in increased tumor cell apoptosis for future treatments of colon cancer. PMID- 10811476 TI - Efficient cotransduction of tumors by multiple herpes simplex vectors: implications for tumor vaccine production. AB - Many gene therapy strategies would be enhanced by efficient transfer of multiple genes into the same cell. Herpes simplex viral amplicon (HSV) vectors are good vehicles for gene transfer because they accommodate large pieces of foreign DNA and transfer genes rapidly and efficiently. The current studies examine whether efficient cotransduction of tumor cells can be accomplished using multiple HSV vectors in a manner useful for clinical gene therapy. Interleukin-12 (IL-12) exists as a heterodimer, with components (m35 and m40) coded for by genes on two separate chromosomes. We constructed HSV vectors carrying either IL12m35 (HSVm35) or IL12m40 (HSVm40) or both genes (HSVm75) separated by an internal ribosome entry site to assess whether gene transfer using a single HSV vector constructed to carry multiple genes has any advantage over gene transfer using multiple vectors that are each carrying single genes. Because IL-12 and IL-2 have been found to have synergistic antitumoral activity, we further analyzed the biologic activity of tumor cells cotransduced by separate HSV vectors carrying genes coding for these two cytokines. The results demonstrate that multiple genes can be inserted into the same cell efficiently using multiple HSV vectors, and that these vectors allow rapid production of tumor vaccines expressing multiple cytokine genes. Thus, gene transfer using HSV may not be limited by the size of the DNA that each vector can accommodate. Immunizations with tumors cotransduced with HSVm35 and HSVm40 were equally effective in eliciting a cytolytic T lymphocyte response and in protecting against tumor growth in vivo as immunization with tumors treated with HSVm75. Immunization with tumors cotransduced with HSVm75 and HSVil2 was superior to immunization with tumors transduced with either alone. The combination of IL-2- and IL-12-secreting tumor cells may be used as an effective immunization strategy against solid tumors. PMID- 10811477 TI - A novel strategy for the tumor angiogenesis-targeted gene therapy: generation of angiostatin from endogenous plasminogen by protease gene transfer. AB - When NIH 3T3 fibroblasts were transduced with a retroviral vector containing a cDNA for porcine pancreatic elastase 1 and cultured in the presence of affinity purified human plasminogen, the exogenously added plasminogen was digested to generate the kringle 1-3 segment known as angiostatin, a potent angiogenesis inhibitor. This was evidenced by immunoblot analysis of the plasminogen digests using a monoclonal antibody specifically reacting with the kringle 1-3 segment, and by efficient inhibition of proliferation of human umbilical vein endothelial cells by the plasminogen digests isolated from the culture medium of 3T3 fibroblasts. However, when Lewis lung carcinoma cells were transduced with the same vector and injected subcutaneously into mice in their back or via the tail vein, their growth at the injection sites or in the lungs was markedly suppressed compared with the growth of similarly treated nontransduced Lewis lung carcinoma cells. Nevertheless, the transduced cells were able to grow as avidly as the control cells in vitro. Assuming that the elastase 1 secreted from the transduced cells is likely to be exempt from rapid inhibition by its physiological inhibitor, alpha1-protease inhibitor, as shown in the inflammatory tissues, the elastase 1 secreted from the tumor cells may effectively digest the plasminogen that is abundantly present in the extravascular spaces and generate the kringle 1 3 segment in the vicinity of implanted tumor cell clusters. Although the selection of more profitable virus vectors and cells to be transduced awaits further studies, such a protease gene transfer strategy may provide us with a new approach to anti-angiogenesis gene therapy for malignant tumors and their metastasis in vivo. PMID- 10811478 TI - Differential expression of a recombinant adeno-associated virus 2 vector in human CD34+ cells and breast cancer cells. AB - The use of autologous hematopoietic stem cell (HSC) grafts after high-dose chemotherapy protocols may be hampered by contamination of the grafts with tumor cells. Because epithelial cells seem to be the natural hosts of adeno-associated virus 2 (AAV-2), we speculated that epithelial tumor cells in HSC grafts might be selective targets for AAV-2-based vectors. To test this hypothesis, the breast cancer cell lines T47D and MCF-7 were infected with a recombinant AAV-2 vector expressing the green fluorescent protein (GFP) gene; in addition, human CD34+ mobilized peripheral progenitor cells were infected with the same vector. At a multiplicity of infection of 100, only 1.39% +/- 0.51% CD34+ cells expressed the GFP gene whereas, 36.06% +/- 6.53% of the infected T47D cells and 41.52% +/- 3.16% of the infected MCF-7 cells expressed the transduced GFP gene. After further optimizing the transduction procedure by using higher multiplicities of infection (100-500) and preincubation of samples with the tyrosine kinase inhibitor genistein, up to 82.52% and 85.35% GFP+ T47D and MCF-7 cells, respectively, were observed. The GFP fluorescence intensity in transduced mammary tumor cells was up to 3 logs higher than that of transduced CD34+ cells. The differential expression of recombinant AAV-2 vectors in hematopoietic and epithelial tumor cells warrants further research with this vector system, including the use of suicide genes for the purging of autologous HSC grafts. PMID- 10811479 TI - CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: A potential tool for gene immunotherapy. AB - Dendritic cells (DC) are the most potent immunostimulatory cells, with the capacity to induce primary T-cell responses. Functional autologous DC can be generated from fetal calf serum-free peripheral blood mononuclear cells in the presence of interleukin-4 and granulocyte-macrophage colony-stimulating factor and are stimulated with a defined cytokine cocktail for terminal maturation. We were able to establish a nonviral transfection protocol for these DC by electroporation. Using enhanced green fluorescent protein as a reporter gene, we achieved transfection efficiencies of up to 10%. FACScan analyses revealed a stable phenotype, and the expression of major histocompatibility complex class II and CD83 was not affected by the transfection conditions used. Like their untransfected counterparts, DC that were functionally transfected with green fluorescent protein were potent inducers of allogeneic T cells. To assess whether cDNAs transfected into DC are functionally expressed, human tyrosinase cDNA was transfected into DC. Tyrosinase-transfected DC, but not controls, resulted in antigen-specific tumor necrosis factor-alpha release of the tyrosinase-specific cytolytic T-cell clone IVSB. Taken together, the data show that genuine (CD83+) mature DC can be transfected using a nonviral method, and that the DC retain their functionality. These DC are ideal candidates for immunotherapy (e.g., cancer therapy). PMID- 10811480 TI - Redirected cellular cytotoxicity by infection of effector cells with a recombinant vaccinia virus encoding a tumor-specific monoclonal antibody. AB - Cytotoxicity is an important function of the immune system that results in the destruction of cellular targets by humoral and/or cellular mechanisms. We wanted to assess the possibility of targeting the lytic function of immune cells toward cancer cells, which express the gene coding for a known tumor antigen (Ag) (GA733 2/epithelial cell adhesion molecule), using a viral vector encoding a monoclonal antibody (mAb) specific for said tumor Ag (CO17-1A). To this end, we have constructed recombinant vaccinia viruses expressing the sequences corresponding to mAb CO17-1A, which recognizes a specific Ag (GA733-2) that is present on the surface of most gastrointestinal carcinomas. The recombinant vectors encoding either a secreted or membrane-anchored form of CO17-1A mAb were used to infect effector cells, which were subsequently assessed for their cytotoxic activity. The recombinant viruses were able to infect both granulocyte-macrophage colony stimulating factor-activated human macrophages and Ag-stimulated murine cytotoxic T lymphocytes. Infected granulocyte-macrophage colony-stimulating factor activated macrophages were found to be able to kill GA733-2-expressing tumor cells. Likewise, infected cytotoxic T lymphocytes, although conserving their original alloreactivity, gained the capability of killing GA733-2-expressing cancer cells. PMID- 10811481 TI - Frape-1 and Frape-3: two different recombinant retroviruses encoding the same human marker gene. AB - A major goal in retroviral-based gene therapy is to establish methods that allow for the selection and tracking of transduced cell populations. Ex vivo gene marking of normal and malignant hemopoietic cells allows the cells to be followed subsequently in vivo. For in vivo applications, a neutral marker gene that is nonimmunogenic is desirable. To track two distinctively treated cell populations in a single individual, we designed and constructed two retroviral vectors; both of these vectors encode a truncated form of the human low-affinity nerve growth factor receptor, a neutral gene that does not transduce signals and is expected to be nonimmunogenic in humans. The two vectors, named Frape-1 and Frape-3, are identical at the protein level but differ at the DNA level, containing restriction sites that allow easy detection by polymerase chain reaction analysis. We show that cell lines and primary CD34+ cells can be readily transduced with these vectors and that transduced cells can be distinguished by polymerase chain reaction- and vector-specific restriction sites. These vectors will be useful for toxicity studies on in vivo gene therapy and for determining the source of relapse in hematological malignancies. PMID- 10811482 TI - Combined chemotherapy of murine mammary tumors by local activation of the prodrugs ifosfamide and 5-fluorocytosine. AB - The success of chemotherapeutic intervention is limited because the necessary high local drug doses cannot be achieved without systemic toxicity. Application of suicide genes (SGs) and direct conversion of prodrugs (PDs) to toxic metabolites in situ by SGs may enhance the efficacy of chemotherapy. To evaluate this strategy in two murine breast cancer models, TS/A and GR, we injected cellulose sulfate capsules harboring cat kidney cells expressing the SGs cytosine deaminase and cytochrome P450 2B1 (CYP2B1) intratumorally. The PDs 5 fluorocytosine and ifosfamide were administered in 3-day intervals. The effect of in situ chemotherapy with each PD alone and the combination was analyzed over a period of 100 days. The results reveal that for TS/A tumors, the antitumoral effect mediated by CYP2B1 is more efficient than that of cytosine deaminase, whereas for GR tumors, both systems worked equally well. Furthermore, we find additive toxicity using both SG/PD systems for both TS/A and GR tumors. PMID- 10811483 TI - Expression of Escherichia coli uracil phosphoribosyltransferase gene in murine colon carcinoma cells augments the antitumoral effect of 5-fluorouracil and induces protective immunity. AB - Uracil phosphoribosyltransferase (UPRT) of Escherichia coli origin can convert 5 fluorouracil (5-FU), a chemotherapeutic agent widely used for solid tumors, to an active intermediate, 5-fluorouridine-5'-monophosphate, as mammalian orotate phosphoribosyltransferase does. To examine whether the E. coli UPRT gene expressed in tumor cells can confer increased sensitivity to 5-FU, we retrovirally transduced Colon 26 cells, a murine colon carcinoma cell line, with the UPRT gene (Colon 26/UPRT cells) and tested the in vivo antitumoral effect of 5-FU in syngeneic immunocompetent mice. After 5-FU administration, tumors of Colon 26/UPRT cells regressed, whereas those of wild-type cells were unaffected. The mice that once eliminated Colon 26/UPRT tumors after 5-FU treatment rejected wild-type cells that were subsequently inoculated but not irrelevant syngeneic tumor cells. This suicide gene/prodrug system was less efficient in nude mice, suggesting that mature alphabeta T cells play a role in the antitumoral effect. The cytotoxicity mediated by the bystander effect was marginal in this system, contrary to the herpes simplex virus-thymidine kinase gene/ganciclovir system. Therefore, expression of the UPRT gene in tumor cells followed by 5-FU administration is a possible strategy for cancer gene therapy, but potentiation of the bystander effect is required for its therapeutic application. PMID- 10811485 TI - NHS remains in the limelight. PMID- 10811484 TI - Ovarian carcinoma cells are effectively transfected by polyethylenimine (PEI) derivatives. AB - As a prerequisite to nonviral gene therapy approaches of ovarian carcinoma, we evaluated the possibility of transfecting established tumor cell lines (SKOV3, IGROV1) as well as primary mesothelial and tumor cells by various polyethylenimine (PEI) derivatives. Several PEI-based vectors were able to effectively transfect these cells, as shown by high luciferase expression levels (10(8) to 10(9) relative light units per milligram of cell protein) that corresponded with 25-50% of green fluorescent protein-positive cells after 24 hours. However, unpredictable differences were observed among the vectors and cell types that a posteriori justified the screening procedure. We also showed that cells that were not transfected after the first experiment remained transfectable in a subsequent transfection experiment to a level similar to that of the initial population. This experiment does not support the emergence of a transfection-resistant cell population and opens the door to multiple therapeutic gene deliveries. Although efficacy and cell targeting still remain to be improved, PEI derivatives appear to be promising molecules for the development of nonviral gene therapy of ovarian carcinoma. PMID- 10811486 TI - After 50 years, the Dutch Cancer Society enters a new era. PMID- 10811487 TI - Health care: a top issue for US Democrats in the election year. PMID- 10811488 TI - World summit against cancer for the new millennium: the Charter of Paris. PMID- 10811489 TI - Estimation of dihydropyrimidine dehydrogenase activity: does it have a role in cancer therapy? PMID- 10811490 TI - Relevance of DNA-ploidy as a prognostic instrument for solid tumors. PMID- 10811491 TI - Germ-cell tumors in childhood and adolescence. GPOH MAKEI and the MAHO study groups. AB - In mature and immature teratoma the treatment is surgical. The risk of recurrence can be estimated from the parameters primary site (with the coccygeal tumors being most at risk), histological grade of immaturity and completeness of the primary resection including the adjacent organ of origin (coccyx, ovary, testis etc.). In case of a microscopically complete tumor resection there is no role for adjuvant chemo- or radiotherapy irrespective of the histological grade of immaturity. Malignant germ-cell tumors (GCT) account for 2.9% of all malignant tumors of children younger than 15 years of age. More than half of the tumors occur at extragonadal sites such as the ovaries (26%), the coccygeal region (24%), the testes (18%) and the brain (18%) represent then primary sites. In patients with extensive tumor growth, metastatic disease or secreting intracranial tumors a delayed tumor resection after preoperative chemotherapy is preferable. In these patients malignant non-seminomatous GCT may be diagnosed clinically due to the increased serum or cerebrospinal fluid levels of the tumor markers AFP and/or beta-HCG. Current risk adapted treatment protocols containing cisplatinum allow long-term remissions in about 80% including patients with bulky or metastatic tumors. In the cisplatinum era the prognostic factors like histology, primary site of the tumor and initial tumor stage have partly lost their former impressive significance in infants and children. On the other hand the completeness of the primary tumor resection according to oncological standards has been established as the most powerful prognostic parameter superior to tumor marker levels or primary site of the tumor. PMID- 10811492 TI - Characterization of dihydropyrimidine dehydrogenase on immunohistochemistry in colon carcinoma, and correlation between immunohistochemical score and protein level or messenger RNA expression. AB - BACKGROUND: Dihydropyrimidine dehydrogenase (DPD) is the first enzyme that metabolizes 5-fluorouracil (5-FU). Until now, enzymatic activity or mRNA expression of DPD has been investigated. However, there are no papers on immunohistochemical evaluation of DPD. We investigated DPD staining on immunohistochemistry, and examined the relationship among immunohistochemical score, protein level and mRNA expression of DPD. MATERIALS AND METHODS: Forty seven resected colon cancer specimens, four colon cancer cell lines, two xenografts by colon cancer cell lines, and human mononuclear cells were used. Immunohistochemistry was performed using DPD monoclonal antibody. Protein levels were determined by Western blot analysis. And mRNA levels were calculated by semi quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: DPD was strongly expressed in the cytoplasm of cancer cells, and in the cytoplasm of macrophage and plasma cells. The immunohistochemical score was more correlated with protein levels (P = 0.0054) than mRNA expression (P = 0.9028). CONCLUSIONS: We investigated the characterization of DPD immunohistochemically, and showed that immunohistochemical expression of DPD can be used to predict the sensitivity of colorectal carcinomas to 5-FU. PMID- 10811493 TI - Randomized study on adjuvant chemotherapy in stage I high-risk ovarian cancer with evaluation of DNA-ploidy as prognostic instrument. AB - PURPOSE: Adjuvant chemotherapy versus observation and chemotherapy at progression was evaluated in 162 patients in a prospective randomized multicenter study. We also evaluated DNA-measurements as an additional prognostic factor. PATIENTS AND METHODS: Patients received adjuvant carboplatin AUC 7 every 28 days for six courses (n = 81) or no adjuvant treatment (n = 81). Eligibility included surgically staged and treated patients with FIGO stage I disease, grade 1 aneuploid or grade 2 or 3 non-clear cell carcinomas or clear cell carcinomas. Disease-free (DFS) and disease-specific (DSS) survival were end-points. RESULTS: Median follow-up time was 46 months and progression was observed in 20 patients in the treatment group and 19 in the control group. Estimated five-year DFS and DSS were 70% and 86% in the treatment group and 71% and 85% in the control group. The hazard ratio was 0.98 (95% confidence interval (95% CI): 0.52-1.83) regarding DFS and 0.94 (95% CI: 0.37-2.36) regarding DSS. No significant differences in DFS or DSS could be seen when the log-rank test was stratified for prognostic variables. Therefore, data from both groups were pooled for the analysis of prognostic factors. DNA-ploidy (P = 0.003), extracapsular growth (P = 0.005), tumor rupture (P = 0.04), and WHO histologic grade (P = 0.04) were significant independent prognostic factors for DFS with P < 0.0001 for the model in the multivariate Cox analysis. FIGO substage (P = 0.01), DNA ploidy (P < 0.05), and histologic grade (P = 0.05) were prognostic for DSS with a P-value for the model < 0.0001. CONCLUSIONS: Due to the small number of patients the study was inconclusive as regards the question of adjuvant chemotherapy. The survival curves were superimposable, but with wide confidence intervals. DNA-ploidy adds objective independent prognostic information regarding both DFS and DSS in early ovarian cancer. PMID- 10811494 TI - Proposing phase I studies: patients', relatives', nurses' and specialists' perceptions. AB - PURPOSE: As of now the primary objective of studies on informed consent in phase I trials has been to assess patients' expectations and reasons for participation. We have previously shown that the quantity of information provided through a procedure of subsequent oral interviews with patients was adequate while the attention paid by the physician to the emotional needs and concerns of patients was not. We wanted therefore to assess and compare the perceptions of the information provided about the investigational study of patients, relatives, the research nurse and the investigator responsible for the phase I trial and the impact this information had on the patients' level of anxiety and depression. PATIENTS AND METHODS: The participation to a phase I study was proposed to patients through two subsequent interviews, the latter attended also by patients' relatives, the research nurse and the investigator coordinating the phase I trial. After the second interview, attendees were requested to complete a questionnaire assessing the principal reason for participating in the study and the informative, emotional and interactive dimension of the information. Patients were also requested to complete the Hospital Anxiety and Depression (HAD) scale before and after the second interview. RESULTS: The completed questionnaires of 31 of 42 patients were retrieved and analysed. The possibility to benefit from the study was indicated as the main reason for participating by 59% of the patients while it was judged to be the case in 78% and 86% of the patients by the nurse and the investigator, respectively. The information was judged to be clear and sufficient in almost all cases by all attendees, while the investigator judged that a lower percentage of patients felt at ease and could express their main worries during the interview, had been helped and were less worried after it than it was judged by the nurse and the relatives. Patients' state of anxiety and depression was not adversely affected by the information provided. CONCLUSIONS: Informing patients on the option of receiving an investigational treatment within a phase I study is feasible and can be done in a way felt appropriate by patients and relatives, nursing and medical professionals. Providing information in an appropriate manner does not increase patients' anxiety and depression. Divergence between the aims and interests of the investigators and patients might explain the difference in the evaluation of physician, a problem which could perhaps be partially overcome by the application of innovative phase I designs. PMID- 10811495 TI - Expression of CD44 variant isoforms and association to the benign form of endocrine pancreatic tumours. AB - BACKGROUND: The expression of CD44 and its isoforms have been shown in many neoplastic tissues to serve as prognostic indicators, therefore, the feasibility of using these as prognostic markers in endocrine pancreatic tumour patients was examined. PATIENTS AND METHODS: Immunohistochemistry (IHC) was performed on 26 tumour samples (5 gastrinomas, 3 glucagonomas, 10 non-functioning tumours, 6 insulinomas, 2 mixed insulinoma and glucagonomas) with monoclonal antibodies against CD44s (standard form) and variant isoforms (v4, v5, v6, v7, v7-8, v9, v10). Staining was correlated to the tumour proliferation, malignancy, metastasis and patients survival. RESULTS: There was variable expression of CD44s. All tumours showed complex expression of many isoforms. CD44v6 and CD44v9 were down regulated in malignant tumours. There was statistical significance of CD44v6 expression in benign tumours (P < 0.05) compared to malignant tumours and near significance in CD44v9 expression (P = 0.0574). Survival of the patients with CD44v6 positive staining was higher than those who were negative (P = 0.0822). Moreover, the expression was well correlated to the patients without any distant metastases (CD44v6, p < 0.001; CD44v9, P < 0.01). Tumour proliferation (Ki67 index) correlated directly to the malignancy (P < 0.05) and there was inverse correlation between Ki67 index and CD44v6 (P < 0.05) as well as v9 (P < 0.05). CONCLUSIONS: Endocrine pancreatic tumours express CD44s and isoforms differentially. Expression of the two isoforms of CD44, namely v6 and v9 seem to be related more to benign form of the tumour and could serve as a predictor of good prognosis. PMID- 10811496 TI - Docetaxel (Taxotere)-cisplatin (TC): an effective drug combination in gastric carcinoma. Swiss Group for Clinical Cancer Research (SAKK), and the European Institute of Oncology (EIO). AB - PURPOSE: A multi-centric trial was performed to explore the clinical activity, in terms of response and toxicity (primary objectives), duration of response and survival (secondary objectives), of docetaxel with cisplatin in advanced gastric cancer (AGC). PATIENTS AND METHODS: Patients with measurable unresectable and/or metastatic gastric carcinoma, performance status < or = 1, normal hematological, hepatic and renal functions and not pretreated for advanced disease by chemotherapy received up to eight cycles of TC (docetaxel 85 mg/m2 dl, cisplatin 75 mg/m2 dl) q3w. Dose escalation to 100 mg/m2 was performed in five patients and was discontinued for excessive toxicity. RESULTS: Forty-eight patients were accrued. A median of 5 cycles/patient was given. We observed 2 complete and 25 partial responses for an overall intent to treat response rate of 56% (95% CI: 41%-71%). Twelve patients had stable disease for > or = 9 weeks (3 cycles). The median time to progression and overall survival were 6.6 and 9 months, respectively. Grade > or = 3 toxicities were neutropenia 81%, anemia 32%, thrombocytopenia 4%, alopecia 36%, fatigue 9%, mucositis 9%, diarrhea 6%, nausea/vomiting 4%, neurologic 2%, and one anaphylaxis precluding treatment administration. We recorded nine episodes of non-fatal febrile neutropenia in eight patients, two of them with docetaxel at 100 mg/m2. There were no direct treatment-related deaths. CONCLUSIONS: TC is active in AGC with a high response rate in a multicentric trial. Despite its hematotoxicity, this regimen is well tolerated and can be recycled as originally planned in 78% of the cases. These results may serve as basis for further developments of docetaxel containing regimens in this disease. PMID- 10811497 TI - Primary node negative breast cancer in BRCA1 mutation carriers has a poor outcome. AB - BACKGROUND: The association between BRCA1 germ-line mutations and breast cancer prognosis is controversial. A historical cohort study was designed to determine the prognosis for women with axillary lymph node negative hereditary breast cancer. PATIENTS AND METHODS: We tested pathology blocks from 118 Ashkenazi Jewish women with axillary lymph node negative breast cancer for the presence of the two common BRCA1 founder mutations, 185delAG and 5382insC. Patients were followed up for a median of 76 months. Somatic TP53 mutations were screened for by immunohistochemistry, and direct sequencing was performed in the BRCA1 positive tumours. RESULTS: Sixteen breast cancer blocks (13.6%) carried a BRCA1 mutation. Young age of onset, high nuclear grade, negative estrogen receptor status and over-expression of p53 were highly associated with BRCA1-positive status (P-values all <0.01). BRCA1 mutation carriers had a higher mortality than non-carriers (five-year overall survival, 50% and 89.6%, respectively, P = 0.0001). Young age of onset, estrogen receptor negative status, nuclear grade 3, and over-expression of p53 also predicted a poor outcome. Cox multivariate analyses showed that only germ-line BRCA1 mutation status was an independent prognostic factor for overall survival (P = 0.01). Among nuclear grade 3 tumours, the BRCA1 mutation carrier status was a significant prognostic factor of death (risk ratio 5.8, 95% confidence interval: 1.5-22, P = 0.009). Sequencing of BRCA1 related breast cancers revealed one TP53 missense mutation not previously reported in breast cancer. CONCLUSIONS: Using a historical cohort approach, we have identified BRCA1 mutation status as an independent prognostic factor for node negative breast cancer among the Ashkenazi Jewish women. Those managing women carrying a BRCA1 mutation may need take these findings into consideration. Additionally, our preliminary results, taken together with the work of others suggest a different carcinogenic pathway in BRCA1-related breast cancer, compared to non-hereditary cases. PMID- 10811498 TI - NCIC-CTG phase II study of gemcitabine in patients with malignant glioma (IND.94). AB - PURPOSE: We conducted a phase II multicentre study of gemcitabine in patients with anaplastic astrocytoma and glioblastoma multiforme at first relapse. PATIENTS AND METHODS: Patients with anaplastic astrocytoma or glioblastoma multiforme receiving a stable dose of steroids and ECOG performance status < or = 3 were eligible for this study at the time of first relapse. One adjuvant chemotherapy regimen was permissible. Patients received gemcitabine 1000 mg/m2 i.v. weekly x 3, repeated on a four-weekly cycle. RESULTS: Of 20 patients enrolled, 15 were evaluable for response, 19 for non-hematological toxicity and 18 for hematological toxicity. Seven patients had anaplastic astrocytoma (AA) and twelve glioblastoma multiforme (GBM). Age ranged from 28-71 years (median 50). Fifteen patients discontinued therapy due to disease progression. The median number of cycles administered was 1 (range 1-11); only two patients received more than three cycles. Hematologic toxicity was acceptable and no grade 4 toxicity was seen. One patient developed Pneumocystis pneumonia and eventual pulmonary embolism; one died of gastric hemorrhage related to steroid therapy. No objective responses were seen. Nine patients had stable disease (median duration 2.7 months, range 0.9-11.2). CONCLUSIONS: Gemcitabine given in this dose and schedule seems well tolerated but is not active in patients with recurrent high-grade gliomas. PMID- 10811499 TI - EGF-related antisense oligonucleotides inhibit the proliferation of human ovarian carcinoma cells. AB - BACKGROUND: The epidermal growth factor (EGF)-like peptides CRIPTO (CR), amphiregulin (AR) and transforming growth factor alpha (TGFalpha) are expressed in human ovarian carcinomas. MATERIALS AND METHODS: The expression of AR, CR and TGFalpha in ovarian carcinoma cell lines was assessed by immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). The antiproliferative effects of antisense phosphorothioate oligodeoxynucleotides (AS S-Oligos) directed against either AR, CR or TGFalpha was evaluated by using a clonogenic assay. RESULTS: A majority of the ovarian carcinoma cell lines was found to express TGFalpha, AR and CR mRNAs and proteins. AS S-Oligos directed against either AR, CR or TGFalpha were able to inhibit the anchorage-independent growth of NIH:OVCAR3 and NIH:OVCAR8 cells in a dose dependent manner. A 30%-50% growth inhibition was observed at a 2 microM concentration of the AS S-Oligos. Treatment of these cells with combinations of EGF-related AS S-Oligos resulted in a more significant growth inhibition when compared to treatment with a single AS S oligo. A 60%-75% growth inhibition was observed using combinations of AR, CR and TGFalpha AS S-oligos at a total concentration of 2 microM. An additive growth inhibitory effect occurred when ovarian carcinoma cells were exposed to the AS S Oligos after treatment with either paclitaxel or cis-platinum. CONCLUSIONS: These data suggest that EGF-related peptides function as autocrine growth factors in ovarian carcinoma cells, and that they might represent targets for experimental therapy of ovarian carcinoma. PMID- 10811500 TI - The urokinase-type plasminogen activator system in resected non-small-cell lung cancer. Rotterdam Oncology Thoracic Study Group. AB - BACKGROUND: Urokinase-type plasminogen activator (uPA), its receptor (uPAR) and plasminogen activator inhibitors (PAI-1 and PAI-2), all play important roles in tumour invasion and metastasis. The tumour levels of the components of the urokinase-type plasminogen activator system (uPA-system) may help to identify individuals with a poor prognosis in postoperative non-small-cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: The levels of uPA, uPAR PAI-1 and PAI-2 were measured by enzyme-linked immunosorbent assay (ELISA) in triton-extracts, prepared from 88 NSCLC tissues (stage I-IIIa) and 74 normal lung tissues from the same patients. RESULTS: The expression levels of uPA, uPAR, PAI-1 and PAI-2 were significantly higher in tumour tissues as compared to their normal equivalents (all, P < 0.0001). Significant relations were found between gender and uPA (P = 0.04) or uPAR (P < 0.001), and between PAI-2 and pathological stage (P = 0.03). For none of the studied factors of the uPA-system a significant relation with survival was found, neither in all patients, nor in the subgroups of patients with squamous-cell lung carcinoma or adenocarcinoma. CONCLUSIONS: The expression levels of the components of the uPA-system were higher in NSCLC tissue as compared to normal lung tissue, but there were no significant relationships between their levels and survival. PMID- 10811501 TI - A phase I study of rhizoxin (NSC 332598) by 72-hour continuous intravenous infusion in patients with advanced solid tumors. AB - BACKGROUND: Rhizoxin (NSC 332598) is a novel macrolide antitumor antibiotic that inhibits microtubule assembly and also depolymerizes preformed microtubules. In preclinical evaluations, rhizoxin demonstrated broad antitumor activity in vitro and in vivo including both vincristine- and vindesine-resistant human lung cancers. Prolonged exposure schedules in xenograft models demonstrated optimal efficacy indicating schedule-dependent antitumor activity. The early phase I and II evaluations a five-minute bolus infusion schedule was studied, however, only modest anti-tumor activity was noted, possibly due to rapid systemic clearance. To overcome these limitations and to exploit the potential for schedule-dependent behavior of rhizoxin, the feasibility of administering rhizoxin as a 72-hour continuous intravenous (i.v.) infusion was evaluated. PATIENTS AND METHODS: Patients with advanced solid malignancies were entered into this phase I study, in which both the infusion duration and dose of rhizoxin were increased. The starting dose was 0.2 mg/m2 over 12 hours administered every 3 weeks. In each successive dose level, the dose and infusion duration were incrementally increased in a stepwise fashion. Once a 72-hour i.v. infusion duration was reached, rhizoxin dose-escalations alone continued until a maximum tolerated dose (MTD) was determined. RESULTS: Nineteen patients were entered into the study. Rhizoxin was administered at doses ranging from 0.2 mg/m2 i.v. over 12 hours to 2.4 mg/m2 i.v. over 72 hours every 3 weeks. The principal dose-limiting toxicities (DLT) were severe neutropenia and mucositis, and the incidence of DLT was unacceptably high at rhizoxin doses above 1.2 mg/m2, which was determined to be the MTD and dose recommended for phase II studies. At these dose levels, rhizoxin could not be detected in the plasma by a previously validated and sensitive high-performance liquid chromatography assay with a lower limit of detection of 1 ng/ml. No antitumor responses were observed. CONCLUSIONS: Rhizoxin can be safely administered using a 72-hour i.v. infusion schedule. The toxicity profile is similar to that observed previously using brief infusion schedules. Using this protracted i.v. infusion schedule the maximum tolerated dose is 1.2 mg/m2/72 hours. PMID- 10811502 TI - Phase I study of Caelyx (doxorubicin HCL, pegylated liposomal) in recurrent or metastatic head and neck cancer. AB - BACKGROUND: Pegylated liposome technology represents a favourable drug-carrier system, since stealth liposomal drugs have a reduced clearance with prolonged circulation half-life and selective drug accumulation in tissues with increased vascular permeability, such as tumor tissues. Caelyx is a pegylated liposome containing doxorubicin, which has been developed to target drug delivery to cancer cells, thus reducing toxicities. Biodistribution studies have shown a selective tumor uptake in patients with advanced head and neck cancer (HNC), thus justifying the present phase I study. PATIENTS AND METHODS: Patients with recurrent or metastatic HNC were treated with Caelyx administered at the starting dose of 30 mg/m2 every three weeks and escalated by 5 mg/m2 per step. Dose escalation was stopped if more than a third of patients of a given cohort had dose-limiting toxicity (DLT), which was defined as grade 4 neutropenia or thrombocytopenia, grade 3 febrile neutropenia, grade 3 thrombocytopenia with bleeding, grade 3 non-hematologic toxicity (except for nausea and alopecia), or > 2 week delay in chemotherapy recycling. The above dose level was defined as maximum tolerated dose (MTD) and the dose level immediately below was recommended for phase II evaluation. Response was evaluated after three courses of chemotherapy. RESULTS: Twenty-four patients were treated at five dose levels. At 50 mg/m2, three out of six patients had grade 3 stomatitis; therefore, this level was defined as MTD and 45 mg/m2 was the selected dose for phase II. Stomatitis occurred in 11 patients across all dose levels, considering all delivered cycles. Neutropenia occurred in 10 of 24 patients, but reached grade 4 in only 2 patients at fourth dose level. Skin toxicity, mainly appearing in the form of palmar plantar erythrodysestesia, was the most frequent toxicity, occurring in 14 patients. Other side effects were mild. One complete response (4%) and seven partial responses (29%) were observed, for an overall response rate of 33% (95% confidence interval (95% CI): 16%-55%). CONCLUSIONS: Caelyx is a safe and promising new treatment in HNC, that deserves further evaluation both alone and integrated within chemo-radiotherapy strategies. PMID- 10811503 TI - Eight-hour infusion versus bolus injection of doxorubicin in the EAP regimen in patients with advanced gastric cancer: a prospective randomised trial. AB - BACKGROUND: Doxorubicin (40 mg/m2/cycle), etoposide (360 mg/m2/cycle) and cisplatin (80 mg/m2/cycle) comprise an efficient regimen in patients with advanced gastric cancer (AGC). However, its excessive hematological toxicity led doctors to avoid using the combination. Doxorubicin is the main cause of myelotoxicity in the EAP regimen. The aim of this study was to compare an eight hour infusion of doxorubicin (arm A) with intravenous injection of doxorubicin (arm B) in the EAP regimen with respect to toxicity, objective responses, time to progression (TTP) and survival in patients with AGC. PATIENTS AND METHODS: One hundred twenty chemotherapy-naive patients with measurable AGC were randomised between September 1994 and August 1998. Sixty patients in arm A and sixty patients in arm B were considered as fully evaluable. The arms were well balanced for age, sex distribution, previous therapy, histological grade and performance status. One-hundred eighty cycles were applied in arm A (median 2) and 201 in arm B (median 4). RESULTS: No difference was detected (P = 0.28) in the response rate of arm A 20% (CR 3; PR 9; 95% CI: 10-30) and B 28% (CR 3; PR 14; 95% CI: 17-40). But there was a significant difference in PD (P = 0.005) between arm A (51%) and arm B (36%). TTP (P = 0.01) and survival (P = 0.02) analyses detected an advantage for arm B vs. arm A. Grades 3-4 toxicity were as follows (arms A%/B%): anemia 8/10, leukopenia 24/26, thrombocytopenia 6/16 (significance, P = 0.05), nausea/vomiting 5/8, diarrhea 6/2, mucositis 8/5. Apart from the trombocytopenia, there was no significant difference in toxicity grades 3-4 between the two arms. Four treatment-related deaths occurred, two in each arm. CONCLUSIONS: Bolus injection of doxorubicin is superior to eight-hour doxorubicin infusion in the EAP regimen in terms of survival, TTP and PD without being significantly more toxic. PMID- 10811504 TI - Pegylated liposomal doxorubicin in treating a case of advanced hepatocellular carcinoma with severe hepatic dysfunction and pharmacokinetic study. AB - BACKGROUND: There is lack of effective and safe chemotherapy for advanced hepatocellular carcinoma. Polyethylene glycol-coated (pegylated) liposomal doxorubicin (PLD) has long circulation time and enhanced drug accumulation in the tumor tissues. It has significant activity in Kaposi's sarcoma, breast and ovarian cancers and the acute adverse effects of free drug are reduced. PATIENTS AND METHODS: A patient with advanced hepatocellular carcinoma was treated with PLD and a pharmacokinetic study was performed. Initial serum total and direct bilirubin were 3.6 and 6.8 folds of upper normal, respectively, and an indocyanine green clearance test at 15 minutes was 26.3% (normal < 15%). RESULTS: Compared to cases with normal liver function, increased volume of distribution of doxorubicin correlated with a large amount of ascites (P < 0.05). The clearance of drug was unexpectedly higher than in cases with normal liver function (P < 0.05). According to the pharmacokinetic studies, the disposition of PLD in this case has not been retarded even in the presence of severe liver dysfunction. Only minimal toxicities including grade 2 stomatitis and moderate leukopenia were observed. The tumor had a partial remission and the patient survived nine months after PLD treatment. CONCLUSION: PLD could serve as a safe and effective treatment for hepatocellular carcinoma even in the presence of impaired liver function. Its role in treating advanced hepatocellular carcinoma is worthy of further study. PMID- 10811505 TI - Endometrial mesodermal mixed tumor occurring after tamoxifen treatment: report on a new case and review of the literature. AB - BACKGROUND: Anti oestrogenic treatment is widely used for breast cancer treatment and prevention of recurrence. Because of concomitant estrogenic effects, tamoxifen exerts carcinogenic properties on the endometrium. Although secondary endometrial cancers usually present as pure adenocarcinomas, other types of rare tumors have also been reported. PATIENTS AND METHODS: Herein we describe the clinical, pathological as well as therapeutic aspects of a new case of endometrial mesodermal mixed tumor occurring after long-term tamoxifen therapy. RESULTS: The present case occured five years after cessation of a five years tamoxifen treatment. The patient failed to respond to doxorubicin and cyclophosphamide when combined to 5-fluorouracil (5-FU), but she reached complete response when the same two drugs were used with carboplatin, suggesting the potential usefullness of platinum derivatives. CONCLUSIONS: A longer latency period might be observed for endometrial mesodermal mixed tumors as compared to adenocarcinomas and could justify a prolonged clinical and ultrasonographic follow-up of patients during and after tamoxifen treatment. When indicated, chemotherapy might require the use of platinum derivatives in this particular type of secondary tumor. PMID- 10811506 TI - Activity of interferon-alpha in relapsed patients with diffuse large B-cell and peripheral T-cell non-Hodgkin's lymphoma. PMID- 10811507 TI - Efficacy of fludarabine and mitoxantrone (FN) combination regimen in untreated indolent non-Hodgkin's lymphomas. AB - PURPOSE: In the last years, fludarabine (FLU) alone or in combination with other drugs has been reported to be effective in the treatment of previously treated low-grade non-Hodgkin's lymphomas (LG-NHL). The aim of this study was to define the therapeutic efficacy and toxicity of a combination of FLU and mitoxantrone (FN regimen) in untreated LG-NHL. PATIENTS AND METHODS: We used a two-drug combination of FLU (25 mg/m2 i.v. on days 1 to 3) and mitoxantrone (10 mg/m2 i.v. on day 1) to treat 27 previously untreated patients with LG-NHL, Chemotherapy was repeated every four weeks for a total of six cycles. Among 27 patients, 17 (63%) were diagnosed with follicular, 6 (22%) with small lymphocytic, and 4 (15%) with immunocytoma subtypes. RESULTS: Of the 27 patients, 18 (67%) achieved complete response (CR) and 6 (22%) partial response, while the remaining 3 (11%) showed no benefit from the treatment. Regarding histology, in the follicular subtype we observed an overall response rate of 94%, with a 76.5% CR rate. The estimated two year relapse-free survival was 83%, and overall survival was 92%. Hematologic grade 3-4 toxicity was seen in only five (3.3%) patients; no opportunistic infections or deaths were associated with the administration of the FN regimen. CONCLUSIONS: These preliminary data show that the FN regimen is a very active, well-tolerated combination chemotherapy for untreated patients with advanced LG NHL. PMID- 10811508 TI - Weekly docetaxel plus gemcitabine or vinorelbine in refractory advanced breast cancer patients: a parallel dose-finding study. Southern Italy Cooperative Oncology Group (SICOG). AB - PURPOSE: The objective of this study was to determine the docetaxel MTD when combined with gemcitabine or vinorelbine in advanced breast cancer patients who had received previous anthracycline-based chemotherapy for advanced disease. PATIENTS AND METHODS: Advanced breast cancer patients aged between 18 and 70 with ECOG PS 0-2 who had not responded to, or had relapsed after, first-line anthracycline-based chemotherapy, were randomized to receive either gemcitabine 1000 mg/m2 or vinorelbine 25 mg/m2 in combination with escalating doses of docetaxel (starting from 30 mg/m2), all on days 1 and 8 every three weeks. Escalation was stopped if > 33% of patients treated at a given dose level showed DLT at the first cycle. RESULTS: A total of 34 patients with locally advanced (8) or metastatic disease (26) were treated, for a total of 94 cycles delivered. Nineteen patients received docetaxel in combination with gemcitabine and 15 with vinorelbine. All patients had been pretreated with anthracyclines, and 24 of 34 had also received weekly dose-dense paclitaxel. A docetaxel dose of 40/m2 proved to be safe when combined on days 1 and 8 with gemcitabine, while a dose of 35 mg/m2 was tolerated in combination with vinorelbine. Overall, nine episodes of DLT, all of them neutropenia, occurred at the first cycle. Considering all 94 cycles, grades 3 or 4 neutropenia and thrombocytopenia occurred in 15 (44%), and 7 (20%) patients. Non-hematologic toxicity was mild, except for three cases of grade 2 peripheral neuropathy. All patients were assessed for response on an 'intent-to-treat' basis. Overall, five partial responses were recorded (docetaxel + gemcitabine = 3 and docetaxel + vinorelbine = 2), for a 15% (95% CI: 5%-31%) overall response rate. Only 1 of 24 (4%) patients who had received weekly dose dense paclitaxel responded to treatment. CONCLUSIONS: The weekly docetaxel administration in combination with either gemcitabine or vinorelbine is a well tolerated treatment for heavily pretreated advanced breast cancer patients. This approach, although sometimes capable of achieving a major response, does not seem advisable in advanced breast cancer patients refractory to both anthracyclines and paclitaxel. PMID- 10811509 TI - MTHFR gene polymorphism and severe toxicity during adjuvant treatment of early breast cancer with cyclophosphamide, methotrexate, and fluorouracil (CMF) PMID- 10811510 TI - Rituximab: active treatment of central nervous system involvement by non Hodgkin's lymphoma? PMID- 10811511 TI - Percutaneous transluminal angioplasty with or without stenting for femoropopliteal occlusions? A randomized controlled study. AB - BACKGROUND: To investigate the-one year outcome of PTA and stenting and PTA alone for femoropopliteal occlusions. DESIGN: Randomized prospective study METHODS: 32 patients with femoropopliteal occlusions were randomized into two treatment groups: PTA and Strecker-stent (n=15) and PTA alone (n=17). The median age of the patients was 71 years. All patients had chronic limb ischaemia, 66% had tissue loss, 19% had rest pain and 15% had disabling claudication. The median ABPI was 0.45. The occlusion was confined to the superficial femoral artery in 30 cases and to the popliteal artery in 2 cases. The median length of the occlusions was 7.3 cm. Aspirin (ASA), 160 mg daily, was administrated postoperatively but no anticoagulation was used. The follow-up included: clinical examination, measurement of ABPI and control angiography at 12 months or earlier when necessary (20 patients). RESULTS: There was no mortality or limb loss as a consequence of the treatment. There were six (16%) immediate major complications in five patients. In the PTA group, one patient had a myocardial infarction and three patients needed arteriography due to bleeding. In the stent group, one patient required arteriography and embolectomy. The one-year mortality was 6% and there were no amputations. Four patients (two in each group) were operated on with a femorodistal bypass. The rate of clinical improvement was 71% after PTA and stent and 60% after PTA alone (p=0.17). An increased ABPI (>0.10) was shown in 50% of the stent group and 61% in the PTA group (p=0.17). Angiographic re occlusions were seen in 33% and 75% in the stent and PTA groups respectively (p=0.17), while the rate of restenosis was significantly higher in the stent group (50% vs 25%) (p=0.033). CONCLUSIONS: Stenting following PTA for femoropopliteal occlusions does not significantly improve neither the clinical state nor the clinical/angiographic patency. The results do not justify any routine placement of stent following PTA in the successfully recanalized femoropopliteal arteries. The low rate of acceptance of a follow-up angiography indicates that this kind of study should preferably use duplex scanning instead of angiography for follow-up. PMID- 10811512 TI - Haemostatic derangement in advanced peripheral occlusive arterial disease. AB - BACKGROUND: Dysbalance of the coagulation and fibrinolysis system was suspected to be a further risk factor for the progression of peripheral occlusive arterial disease (POAD). Reports on disturbed platelet function in advanced disease, however, were contradictory. Therefore, we studied haemostasis parameters and platelet function in symptomatic patients with peripheral arterial disease. METHODS: 60 peripheral arterial disease patients hospitalised for invasive diagnostic procedures were included into this comparative study. Patients were clinically stratified according to the criteria for chronic limb ischemia (grade I: n=36; grade II: n=11; grade III: n=13). Plasma fibrinogen, antithrombin III, von Willebrand factor, tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1) prothrombin time, and activated partial thromboplastin time were determined using standard methods. We measured flow cytometrically, the platelet activation marker P-selectin on nonstimulated, ADP- and TRAP-6 stimulated platelets. Angiographic data were assessed using the Bollinger score. RESULTS: Plasma levels of the procoagulant proteins fibrinogen (grade I: 3.7/grade II: 3.9/grade m: 4.0 g/l) and vWF (158/156/178%) increased and of antithrombin III (109/103/102%) and the PAI-1/tPA ratio (5.2/5.0/4.1) decreased with progressive disease. Highest platelet activation levels were observed in the CLI grade II subgroup. A significant correlation of disease severity was seen with the ankle-brachial pressure index (p=0.006; r=0.39) and with the Bollinger score (p=0.002; r=-0.41). CONCLUSIONS: Progressive peripheral obstructive arterial disease was associated with platelet hyper-reactivity, haemostatic dysbalance of pro- and anticoagulant proteins, and a counterregulatory increase of fibrinolytic activity. Therapeutic concepts should include these pathogenetic mechanisms. PMID- 10811513 TI - Intravascular ultrasound observations during iliac stent deployment. AB - BACKGROUND: To evaluate the role of intravascular ultrasound (IVUS) during iliac stent deployment, with comparison of four major types of iliac stents. METHODS: Thirty-eight iliac arteries of 37 patients were observed with intravascular ultrasound after implantation of various stents including Palmaz stents in 10, Memotherm stents in 11, Wallstent in 10, and Strecker stents in 7. Quantitative measurements on ultrasound included the ratio of the short-axial to the long axial diameters of the stent (symmetry index), the ratio of stent cross-sectional area to that of the reference lumen (expansion index), and stent-to-wall apposition. RESULTS: Intravascular ultrasound revealed significant differences among four major types of iliac stent, in spite of satisfactory angiographic appearances in all patients. It demonstrated significant deformity of the Strecker stent (symmetry index of 0.76-0.09) compared with other stents. The Memotherm stent and the Palmaz stent were superior to other stents in terms of degree of expansion (mean expansion index of 0.87 and 0.82 respectively). Stent cross-sectional area greater than 80% of the reference lumen could be sufficient for iliac stent deployment. The Palmaz stent was superior to other stents in terms of stent-to-wall apposition. CONCLUSIONS: Intravascular ultrasound can provide precise and useful cross-sectional morphological and quantitative information in terms of stent configuration, degree of stent expansion, and stent to-wall apposition. PMID- 10811514 TI - Evaluation of blood flow velocity changes in middle cerebral artery from contralateral transorbital approach as the alternative method of monitoring during carotid surgery in patients with no temporal acoustic "window". AB - BACKGROUND: We evaluated the possibility of utilising the transorbital approach for middle cerebral artery (MCA) blood flow velocity monitoring as the alternative method of intraoperative evaluation of cerebral ischaemia risk in patients with no temporal acoustic "window". METHODS: We insonated the MCA from contralateral, transorbital approach at a depth ranging from 68 to 80 mm. Ten patients with no temporal window (group A) undergoing carotid surgery were examined intraoperatively by means of this method. RESULTS: For comparison a reference group of 15 patients with temporal acoustic windows present (group B) was established. In this group, the simultaneous (double) records of MCA blood flow velocity changes from transtemporal and transorbital approaches during carotid endarterectomy (CEA) were conducted. Mean blood flow velocity (Vmean) was recorded after anaesthesia, during a 120-sec clamp test, after shunt insertion (if necessary), immediately after and 5 min after clamp release. CONCLUSIONS: We would recommend transorbital Doppler monitoring as a method of choice when the transtemporal approach is not possible. Although quite difficult, this method appears to be safe and reliable in the intraoperative evaluation of patients at risk of cerebral ischaemia. PMID- 10811515 TI - Postcarotid endarterectomy symptoms. Pre- and postoperative clinical and MRI findings. AB - BACKGROUND: The hyperperfusion syndrome represents a highly debated clinical entity, without having yet any clinically identifiable limits. The correlation of clinical with pre- and postoperative magnetic resonance imaging (MRI) findings following carotid endarterectomy in patients with a possible hyperperfusion syndrome was investigated. METHODS: DESIGN: Prospective clinical and laboratory study. SETTING: At the Aretaieio University Hospital. MEASURES: Comparison to the postoperative clinical symptomatic and MRI findings. Patients. We studied 30 patients (mean age 66.6) of whom 14 (46.6%) were asymptomatic, 16 (53.4%) were symptomatic, 17 (56.6%) were hypertensive, 18 (60%) hyperlipidaemic, 5 (16.6%) diabetic and 17 (56.6%) had coronary artery disease. RESULTS: Twenty-one patients (70%) exhibited disturbances such as ipsilateral headache, seizures, vomiting or facial pain; immediate postoperative MRI scans revealed new ischaemic foci in 4 (19%) without any objective neurological findings. Lesions displayed in postoperative MRI scans did not always cause symptoms and normal findings on MRI did not exclude the presence of symptoms. Hypertension favoured the presence of subjective disturbances, without necessarily any change in the MRI appearances. Preoperative MRI and clinical findings did not contribute to the identification of patients who might have exhibited postoperative symptoms. Thus, the symptomatology remains unclear and open to debate, as a variety of other conditions may produce the same clinical picture postoperatively. CONCLUSIONS: Subjective disturbances, objective neurological findings and MRI abnormalities with or without hypertension may appear independently after carotid endarterectomy, a fact which makes the pathogenesis of the hyperperfusion syndrome more difficult to elucidate. PMID- 10811516 TI - Comparison of near-infrared spectroscopy with air plethysmography in detection of deep vein thrombosis. AB - BACKGROUND: As a noninvasive screening test, air plethysmography (APG) is a reliable and frequently used modality in the detection of deep vein thrombosis (DVT). Although APG is highly sensitive for the diagnosis of proximal DVT, its sensitivity for identifying calf DVT is poor. Near-infrared spectroscopy (NIRS) is a new modality which can be used to evaluate venous retention during walking. The purpose of this study was to investigate the diagnostic value of NIRS for the detection of DVT, particularly isolated calf DVT, in comparison with APG. METHODS: Fifty limbs of 39 consecutive patients with clinically suspected DVT were studied. All patients were examined by venography and APG. Patients also underwent a treadmill-walking test with simultaneous NIRS. Deoxygenated hemoglobin was continuously measured by NIRS during exercise. The ambulatory venous retention index obtained from serial DeoHb changes was calculated in each patient. RESULTS: Venography demonstrated DVT in 35 limbs, confined to the calf in 9 limbs, and with the involvement of a proximal deep vein with thrombi in 26. The overall sensitivity of NIRS and APG was 97% (34/35) and 80% (28/35), respectively. NIRS was more sensitive than APG for detecting isolated calf DVT (89% [8/9] and 22% [2/9], respectively), while both tests identified proximal DVT in all limbs. CONCLUSIONS: NIRS is a highly sensitive method for the diagnosis of haemodynamically significant DVT. NIRS may become a useful screening test because of its reliability in detecting calf vein thrombi that cannot be identified by APG. PMID- 10811517 TI - Postmastectomy lymphoedema. Treatment and a five-year follow-up study. AB - BACKGROUND: Lymphoedema of the arm is a common problem after mastectomy and radiotherapy of mammary cancer. In a prospective 5-year follow-up study we investigated the development of arm oedema and the effect of conservative compression treatment. METHODS: Two hundred and twenty-six patients who had undergone mastectomy were investigated. Oedema formation was estimated by recording displaced water volume of both arms preoperatively and six times post operatively. One hundred fifty-seven patients (70%) could be followed for five years. An inter-arm difference of 100 ml or more was defined as oedema. Three types of treatment were given: (1) Compression with stockings in 28 patients, (2) intermittent compression (Flowtron) in 8 and (3) intermittent compression (Lympha Press) and compression sleeves in 19 patients. RESULTS: A total of 46 patients (20%) developed post-operative oedema, 17 within six months and 29 one year postoperatively. The average oedema volume was 418 ml. Most patients with a moderate or severe oedema had irradiation therapy. Severe lymphoedema (750 ml or more) occurred in patients with irradiation postoperatively and lymph node engagement. There was no correlation between patient age and oedema formation. Compression therapy with stockings reduced oedema in 15 out of 28 patients (54%) and prevented further swelling. Therapy with stockings and Lympha Press reduced the arm volume in 13 out of 19 patients (68%). Four out of eight patients treated with Flowtron showed oedema reduction. In most patients cessation of treatment resulted in relapse of swelling to the same degree as before. CONCLUSIONS: Compression therapy is beneficial to control postmastectomy arm swelling. This applies to different modalities and prolonged periods of treatment are required to check progression. Application of stockings is the simplest way to treat postoperative oedema. In most cases postoperative oedema appeared during the first year after surgery and the most severe cases occurred after irradiation. PMID- 10811518 TI - The Friday afternoon pulmonary angiogram: temporal relationship between onset of symptoms and requests for angiography. AB - BACKGROUND: To determine if pulmonary embolism symptoms and pulmonary angiography have a proclivity for a particular day of the week, month or season (Part I) and to assess the length of time elapsing between the appearance of symptoms and requests for pulmonary angiography (Part II). DESIGN: Prospective (in part) collection of data and retrospective review of clinical records. SETTING: Large metropolitan, university affiliated hospital. METHODS: Part I: 2,969 consecutive pulmonary angiograms performed over 17 years, were collected according to the day of the week, month and season of their performance. Relevant rates were calculated. Part II: A study of the time interval between the onset of pulmonary embolism symptoms and requests for and performance of angiography was conducted in 128 patients. Results. Part 1. In the course of 17 years, more pulmonary angiograms were performed on Fridays (mean 37.2, SE 1.8), than on any other day of the week (p <.001). No proclivity for a particular month or season was observed. Part II. The mean time elapsed between the onset of symptoms and requests for angiography was 24 hours (range 0.2-162, SE 4.3) for inpatients, and 45 hours (range 3-358, SE 7.8) for outpatients. The angiogram was completed urgently, within two hours of the onset of symptoms, only in 2.4%. CONCLUSIONS: Pulmonary angiography has a proclivity for Fridays. Pulmonary embolism symptoms and consequent angiograms do not favour a specific day, month or season. In most cases the time elapsing between the onset of symptoms and requests for angiography is too long to be consistent with urgency. The frequent Friday pulmonary angiogram reflects a tendency to "mop up" unfinished business prior to the week's end. PMID- 10811519 TI - Effect of Ruscus extract and hesperidin methylchalcone on hypoxia-induced activation of endothelial cells. AB - BACKGROUND: Ruscus aculeatus extract and the flavonoid hesperidin methylchalcone (HMC) are drugs used in the treatment of chronic venous insufficiency. METHODS: In the present study, we investigated their effects on the activation of endothelial cells by hypoxia, a condition which mimics venous blood stasis. RESULTS: We observed that Ruscus extract was able to inhibit the activation of endothelial cells by hypoxia: the decrease in ATP content, the activation of phospholipase A2 as well as the subsequent increase in neutrophil adherence with a maximal protection obtained at 50 microg/ml. HMC was also able to inhibit the hypoxia-induced decrease in ATP content. Furthermore, the effects of Ruscus extract and of HMC on this decrease seem to be additive. CONCLUSIONS: The biochemical mechanism evidenced in this work might explain some of the beneficial therapeutic effects of these products in the treatment of chronic venous insufficiency patients. PMID- 10811521 TI - Sodium/hydrogen ion exchange inhibitor ameliorates ischaemia-reperfusion injuries in the rat hind limb. AB - PURPOSE: This study was designed to investigate the effect of a newly synthesised sodium/hydrogen ion exchange inhibitor, SM-20550, on ischaemia-reperfusion induced injury in a rat hind limb model. METHODS: In order to induce ischaemia of the hind limbs, the abdominal aorta just distal to the renal arteries and the bilateral femoral arteries were clamped. Nineteen rats were divided into three groups. In the sham group (n=5), the vessels were only dissected and a vehicle solution was administered. In the control group (n=7), the vessels were clamped for five hours, and a vehicle solution was administered 10 minutes prior to clamping and continued for five hours after reperfusion. In the SM group (n=7), clamping was maintained for five hours with a bolus injection of SM-20550 and continuous infusion of the solution for five hours after reperfusion. Water content of the left gastrocnemius muscle was calculated. The right gastrocnemius was fixed in 10% formalin. A transverse thin section was stained with antimyoglobin antibody. Stained cells of the right gastrocnemius were counted and the myoglobin staining index was calculated. RESULTS: Water content was significantly (p<0.002) lower in the SM group than in the control group. The myoglobin staining index was significantly (p<0.01) higher in the SM group than in the control group. There was no significant difference between the control and the SM groups in creatinine phosphokinase (CPK) and lactate dehydrogenase (LDH). CONCLUSIONS: Our results indicate that the sodium/hydrogen ion exchange inhibitor, SM-20550, ameliorates oedema formation and ischaemia-reperfusion induced injury of the skeletal muscle. PMID- 10811520 TI - The effect of four-year hypolipidaemic treatment on the intimal thickness of the common carotid artery in patients with familiar hyperlipidaemia. AB - BACKGROUND: Cholesterol lowering in patients with above-average cholesterol levels has been shown to reduce the progression of atherosclerosis. We assess the effects of lipid lowering therapy on the progression of early, preintrusive carotid arterial atherosclerosis in high risk patients with familial hyperlipidaemia free of symptomatic cardiovascular disease. METHODS: Fifty-two patients with familial hyperlipidaemia by were treated by diet and various hypolipidaemic drugs. Eighteen individuals were not taking hypolipidaemic drugs. In a prospective study by B-mode ultrasound we assessed the intima-media thickness of the distal common carotid arterial (CCA) far wall at baseline and after 4 years. RESULTS: In a subgroup of 25 patients with familial hypercholesterolaemia there was a significant decrease in total and LDL cholesterol and reduction in the intima-media thickness (IMT) of the common carotid artery from 0.78+/-0.22 mm to 0.69+/-0.17 mm (p=0.004). In a subgroup of 27 patients with familial combined hyperlipidaemia significant decreases in total and LDL cholesterol and triglycerides were associated with a decrease in the IMT of common carotid. artery from 0.72+/-0.22 mm to 0.67+/-0.15 mm (p=0.044). In 18 individuals, who were not taking hypolipidaemic drugs, there were no significant changes in the levels of cholesterol and triglycerides and in the IMT of the common carotid artery (increase from 0.58+/-0.18 mm to 0.62+/-0.13 mm, p>0.05). CONCLUSIONS: Lipid-lowering therapy in patients with familial hyperlipidaemia free of symptomatic cardiovascular disease reverses the progression of early, preintrusive atherosclerosis of the carotid artery. It is a beneficial sign indicating the possibility for atherosclerosis regression. PMID- 10811522 TI - Therapeutic efficacy of flavonoids in oedema following reperfusion on acutely ischaemic legs. AB - BACKGROUND: The aim of this study was to investigate the effect of a purified, micronised, flavonoid fraction (Daflon) in lower limb oedema developed after delayed embolectomy and successful reperfusion of acutely ischaemic legs. METHODS: Our series consisted of 19 patients with prolonged, acute ischaemia of the lower extremity caused by arterial embolism, who had undergone successful embolectomy. Patients were randomised into two groups; ten patients (group I) treated with flavonoids, were compared to nine given a placebo. Ankle and calf circumferences, venous capacitance and venous emptying time were measured preoperatively [T1], on the 2nd [T2], 5th [T3], 8th [T4] and 30th [T5] postoperative days. RESULTS: An increase of ankle and calf circumference up to 9% and 13.5% respectively, as compared with preoperative values was demonstrated. There was a reduction in oedema formation in patients with the Daflon group, which reached statistical significance only in the ankles (p=0.0276). Calf differences were statistically significant on the second and fifth postoperative days (p<0.05). Venous haemodynamics were considerably improved by Daflon (p<0.001). CONCLUSIONS: Daflon 500 mg had an inhibitory effect on moderate oedema developed after revascularisation of an ischaemic limb. PMID- 10811523 TI - Effect of glycated low density lipoprotein on smooth muscle cell proliferation. AB - BACKGROUND: We investigated the effect of glycated low density lipoprotein (LDL) on smooth muscle cell proliferation. METHODS: Blood was drawn from 6 healthy subjects after overnight fasting. Native LDL was obtained by separating LDL from the samples with sequential ultracentrifugation. Glycated LDL was prepared by glycating the native LDL in vitro. Native and glycated LDL were added to a medium containing cultured porcine coronary artery smooth muscle cells, and the change in cell proliferation was examined after 24, 48, 72, and 96 hs. The cells were counted using a cell counting kit (Dojin Chemical Co., Ltd.). RESULTS: There was no significant difference in the cell count between the control group, in which only PBS was added, and the native LDL group. However, cell proliferation was appreciably higher in the glycated LDL group than in the native LDL group. The mean total cell count at 24, 48, 72 and 96 hs was significantly higher (p<0.01) in the glycated LDL group (median: 0.843; range: 0.576-1.060) than in the native LDL group (median: 0.541; range: 0.282-0.683). CONCLUSIONS: These findings suggest that glycated LDL induces significantly greater acceleration of smooth muscle cell proliferation than does native LDL. Therefore, the acceleration of smooth muscle cell proliferation requires modification of LDL. PMID- 10811524 TI - Iloprost in the management of leg ulcer in polyarteritis nodosa. A case report. AB - The authors report the case of a patient with a large leg ulcer secondary to polyarteritis nodosa unresponsive to corticosteroid, cytotoxic, and anticoagulant therapy who was successfully treated with iloprost. PMID- 10811525 TI - An open-label, randomised multicentre study comparing the efficacy and safety of CYCLO 3 FORT versus hydroxyethyl rutoside in chronic venous lymphatic insufficiency. AB - BACKGROUND: The present study was designed to compare the safety and efficacy of a combination of Ruscus aculeatus, hesperidin methyl chalcone and ascorbic acid (CYCLO 3 FORT) versus that of hydroxyethyl rutoside in the treatment of chronic venous lymphatic insufficiency. METHODS: This open-label, randomised multicentre study was conducted on outpatients treated for 90 days. The patients were from three different regions of Argentina. Eighty patients, men and women, 30 to 70 years of age, with symptoms of chronic venous lymphatic insufficiency, i.e. heavy, tired, swollen, or painful legs were enrolled and assigned to two groups: one group treated with CYCLO 3 FORT, the other with rutoside. The symptoms were assessed at baseline, and after 30, 60, and 90 days. The patient himself was asked to assess subjective symptoms, the physician measured the size of the affected limbs. The efficacy was rated on a 3-point scale and safety was evaluated by the number of side effects. RESULTS: The result data show that when all patients were comparable at baseline, after 90 days, those treated with CYCLO 3 FORT reported more rapid and more complete regression of symptoms than those in the rutoside group, a statistically significant difference (p<0.01). Likewise, a significant reduction in affected limb size was observed in both groups but persisted after 90 days in the CYCLO 3 FORT group only (p<0.01). CONCLUSIONS: As a conclusion : CYCLO 3 FORT was safe and more effective than rutoside in the treatment of venous lymphatic insufficiency. And the CYCLO 3 FORT efficacy is probably associated with its unique mechanism of action. PMID- 10811526 TI - Surgical management of ascending saphenous thrombophlebitis. AB - BACKGROUND: Acute saphenous vein ascending thrombophlebitis is recognised to be a dangerous condition due to the reported high incidence of deep vein thrombus involvement and possibly fatal pulmonary embolism. We assessed the accuracy of duplex scanning in determining the extent of thrombosis as well as the effectiveness of surgical treatment. METHODS: We retrospectively reviewed 146 patients referred to our Vascular Laboratory for acute superficial thrombophlebitis from 1987 to 1997. Duplex scanning identified 85 cases of superficial thrombophlebitis involving at least a segment of the saphenous vein localised below the knee (58.2%); 37 of thrombophlebitis extending into both the superficial and deep venous systems (25.3%), and 24 of saphenous thrombosis extending to within 5 cm of the saphenofemoral junction (16.4%). The latter group underwent saphenofemoral disconnection. We compared the preoperative duplex with the surgical reports and evaluated the surgical results. RESULTS: We did not observe any complication. Return to work and normal activity occurred within 3-5 days. When varicose vein thrombectomy was performed concurrently, the patients had better postoperative pain control. CONCLUSIONS: Duplex scanning showed 100% accuracy both in determining the presence of thrombosis and its extent. Saphenofemoral disconnection for thrombosis involving the saphenofemoral junction is a safe procedure and can be performed on an outpatient basis. PMID- 10811527 TI - Mechanism underlying increased platelet reactivity in patients with peripheral arterial disease. PMID- 10811528 TI - The proinflammatory transcription factor NFkappaB: a potential target for novel therapeutical strategies. AB - NFkappaB is a ubiquitous transcription factor involved in the proinflammatory response to cytokines (such as IL-1 or TNF-alpha) and some particular stresses. The unraveling of the molecular mechanisms involved in its activation is quite recent, particularly in the case of IL-1-stimulated cells, but the identification of the molecules involved in this pathway opens new prospects in both pharmacological and toxicological research. PMID- 10811529 TI - Proteins of the Ras pathway as novel potential anticancer therapeutic targets. AB - Ras proteins are molecular switches that constitute a pivotal element in the control of cellular responses to many incoming signals, and in particular mitogenic stimulations. They act through multiple effector pathways that carry out the biological functions of Ras in cells. Since mutations that constitutively activate Ras proteins have been found in a high proportion of human malignancies and participate in oncogenesis, a number of therapeutic anticancer strategies aimed against the activity or action of Ras proteins have been developed. This paper reviews the principal aspects of the Ras signaling pathway and describes some of the attempts to develop antitumor drugs based on this concept. PMID- 10811530 TI - New approaches to estimating the mutagenic potential of chemicals. AB - New developments in mutagenic risk assessment have appeared in the past few years. New methods have been developed such as in vitro micronucleus assay for chromosomal alterations, comet assay for primary DNA damage, use of transgenic animals to detect in vivo gene mutations, and fluorescent in situ hybridization method to detect aneuploidy. Other new methods will be developed in the few next years, including the use of DNA chips and the use of molecular biological methods. Several micromethods have been developed to test a great number of chemical compounds. New concepts have appeared concerning interpretation of data, and particularly of thresholds especially in the case of aneugens; in some cases metabolic or mechanistic thresholds were demonstrated. Genotoxic studies are best integrated into toxicological testing: for example, some genotoxicity tests can be integrated into subacute toxicology; interpretation of data includes metabolism; and toxicokinetic data relate to other toxicological studies. Conversely, genotoxicity data can be used to interpret toxicology studies. PMID- 10811531 TI - Hepatotoxicity due to mitochondrial dysfunction. AB - Mitochondria are involved in fatty acid beta-oxidation, the tricarboxylic acid cycle, and oxidative phosphorylation, which provide most of the cell energy. Mitochondria are also the main source of reactive oxygen species in the cell and are involved in cell demise through opening of the mitochondrial permeability transition pore. It was therefore to be expected that mitochondrial dysfunction could be a major mechanism of drug-induced liver disease. Microvesicular steatosis (which may cause liver failure, coma, and death) is the consequence of severe impairment of mitochondrial beta-oxidation. Endogenous compounds (such as cytokines or female sex hormones) or xenobiotics (including toxins such as ethanol and drugs such as aspirin, valproic acid, ibuprofen, or zidovudine) can inhibit beta-oxidation directly or through a primary effect on the mitochondrial genome or the respiratory chain itself. In some patients, infections and cytokines, or inborn errors of beta-oxidation enzymes or the mitochondrial genome, may favor the appearance of drug-induced microvesicular steatosis. Nonalcoholic steatohepatitis may develop under conditions causing prolonged, microvesicular, and/or macrovacuolar steatosis. In this condition, chronic impairment of mitochondrial beta-oxidation (causing steatosis) and the respiratory chain (increasing the production of ROS) lead to lipid peroxidation, which, in turn, may cause the diverse lesions of steatohepatitis, namely, necrosis, inflammation, Mallory's bodies, and fibrosis. Finally, mitochondria are involved in several forms of drug-induced cytolytic hepatitis, through inhibition or uncoupling of respiration or through a drug-induced or reactive metabolite induced mitochondrial permeability transition. The latter effect commits hepatocytes to either apoptosis or necrosis, depending on the number of organelles that have undergone the permeability transition. PMID- 10811532 TI - Airborne particles evoke an inflammatory response in human airway epithelium. Activation of transcription factors. AB - PM10, the commonly used indicator of respirable environmental suspended particulate matter with a mean aerodynamic diameter of less than 10 microm, is composed of organic or elemental carbon aggregates containing various metals, acid salts, organic pollutants (polyaromatic hydrocarbons, quinones, nitroaromatic hydrocarbons, etc.), and biological contaminants. In urban and industrial areas, fossil fuel combustion products (e.g., diesel exhaust particles and residual oil fly ash) are the main contributors to PM10. Epidemiological data show that air pollution particulates cause adverse pulmonary health effects, especially in individuals with preexisting lung diseases. A critical cell type that encounters particles after inhalation and that is affected in a number of respiratory diseases is the epithelial cell of the airway and alveoli. In vitro studies have shown that PM10 is responsible for the production and the release of inflammatory cytokines by the respiratory tract epithelium as well as for the activation of the transcription factor NFkappaB. As many of the adsorbed materials on the particle surface are direct oxidants (metals, quinones) and indirectly produce reactive oxygen species, it is hypothesized that oxidative stress may be a component of the mechanisms by which particles activate cytokine production and NFkappaB in epithelial cells. PMID- 10811533 TI - A reappraisal of the role of Zn2+ in TGF-beta1-induced apoptosis in primary hepatocytes. AB - There is now a wealth of information regarding the apoptotic mode of cell death and its importance in toxicological studies in many mammalian organs including the liver. In this study, we investigated the modulatory effects of the heavy metal Zn2+ on transforming growth factor-beta1 (TGF-beta1)-induced apoptosis in primary rat hepatocytes. Apoptosis induced by TGF-beta1 (1 ng/ml) in hepatocytes was accompanied by nuclear condensation as assessed morphologically by staining with Hoechst 33258 and DNA cleavage as detected biochemically by in situ end labeling, field inversion and conventional gel electrophoresis. Pretreatment with 100 micromol/L Zn2+ abrogated the nuclear condensation, in situ end-labeling, and DNA laddering in TGF-beta1-treated hepatocytes. Surprisingly, Zn2+ did not inhibit the formation of high-molecular-weight DNA fragments (30-50 kbp to 250 300 kbp). These data provide evidence that Zn2+ exerts its effects on the endonucleases that act downstream in the execution phase of TGF-beta1-induced apoptosis in hepatocytes. PMID- 10811534 TI - Increased expression of hepatic DNA methyltransferase in smokers. AB - The DNA methyltransferase enzyme (DNA MTase) catalyzes DNA methylation at cytosines in CpG dinucleotides. 5-Methylcytosine modification of DNA is important in gene regulation, DNA replication, chromatin organization and disease. Increased levels of DNA MTase have been associated with the initiation and promotion of cancer. This study was conducted to assess whether cigarette smoking and other factors, such as age and gender, influence DNA MTase expression in nontumorous tissue. DNA MTase was significantly (p<0.05) higher in samples from cigarette smokers; the mean level of DNA MTase mRNA was almost 2-fold higher in these samples than in those from nonsmokers. Levels of DNA MTase mRNA were higher in samples from females than in those from males, but the difference was not statistically significant. Age was not associated with DNA MTase levels. Increased levels of DNA MTase in individuals who smoke may indicate a greater susceptibility to the risk of cancer since increased levels of this enzyme are found in cancer cell lines and human tumors. The results of this study suggest that further investigations of increased expression of this enzyme as a predisposing factor for cancer susceptibility are needed. PMID- 10811535 TI - Differential toxicity of anticancer drugs on late (GM-CFC) and early (LTC-IC) hemopoietic progenitors in vitro. AB - The clinical hematological toxicity of cytotoxic drugs can be acute, with a nadir of neutrophil count after 2 weeks and recovery the following week, or subacute, with a nadir of neutrophil count after 3 weeks and recovery in the following 2-3 weeks. The explanation usually given for this difference is that drugs in the first group are more toxic to mature hemopoietic precursors, while drugs of the second type are more toxic to undifferentiated cells. In an attempt to verify this hypothesis, we compared in vitro the effect of toxic doses of etoposide and tallimustine as representatives of drugs with acute toxicity, and of BCNU, melphalan, and carzelesin as representatives of drugs with subacute toxicity. Their effects were studied separately on more differentiated and earlier progenitors represented by granulocyte-macrophage colony-forming cells (GM-CFC) and long-term culture-initiating cells (LTC-IC), respectively. Etoposide, melphalan, BCNU, and carzelesin showed higher toxicity in differentiated than in early precursors: the concentration of drug inhibiting 70% (ID70) of GM-CFC inhibited only by 10-40% the growth of LTC-IC. Tallimustine, in contrast, inhibited both GM-CFC and LTC-IC at comparable levels. These results do not correspond to the clinical pattern of myelotoxicity observed for those drugs. We conclude that the differential effects of antitumor drugs on later (GM-CFC) or earlier (LTC-IC) hemopoietic precursors may not represent a valid model for the pattern of myelotoxicity observed in humans. PMID- 10811536 TI - The nephrotoxin ochratoxin A induces apoptosis in cultured human proximal tubule cells. AB - To test the apoptotic potential of the nephrotoxic mycotoxin ochratoxin A (OTA), we exposed human proximal tubule-derived cells (IHKE cells) for various times to OTA concentrations close to those occurring during dietary exposure (from 2 to 100 nmol/L) and investigated caspase 3 activation, chromatin condensation, and DNA fragmentation. OTA induced a time- and concentration-dependent activation of caspase 3: concentrations as low as 5 nmol/L OTA caused a slight but significant increase in caspase 3 activity after 7 days of OTA exposure. Exposure to 10 nmol/L OTA for 72 or 24 h led to a significantly increased activity of caspase 3 in human proximal tubule-derived cells. Radical scavengers such as N acetylcysteine had no effect on OTA-induced caspase 3 activation. Chelation of intracellular calcium with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethylester) (BAPTA-AM) also showed no effect. Exposure to 30 nmol/L or more OTA led to DNA fragmentation and chromatin condensation in IHKE cells. Cultured renal epithelial MDCK-C7 and MDCK-C11 or OK cells also showed increased caspase 3 activity after OTA exposure. We conclude that exposure to low OTA concentrations can lead to direct or indirect caspase 3 activation and subsequently to apoptosis in cultured human proximal tubule cells and in other renal epithelial cell lines of different origins. PMID- 10811537 TI - Estrogen replacement therapy and MRI-demonstrated cerebral infarcts, white matter changes, and brain atrophy in older women: the Cardiovascular Health Study. AB - OBJECTIVE: We studied the relationship between the use of estrogen replacement therapy (ERT) and cerebral magnetic resonance imaging (MRI) abnormalities among older women. DESIGN: A population-based prospective study (Cardiovascular Health Study). SETTING: Four regions in the United States. PARTICIPANTS: A total of 2133 (62.9% of the eligible) women aged 65 to 95 years (mean age 74.8), on whom MRI was performed in 1992-1994. MEASUREMENTS: Presence of global brain atrophy, white matter changes, small infarct-like lesion (ILL) (<3 mm), MRI infarcts (> or =3 mm, mostly small and asymptomatic), and cognitive function as measured by Mini Mental State Exam (MMSE), and by ERT use (current/past/never), adjusted for a number of socioeconomic, lifestyle, and reproductive covariates. RESULTS: Current use of ERT was reported by 15% and past use by another 23% of participants; 35% of all women had MRI infarcts. The prevalence of MRI infarcts did not differ in current or past users from those who had never used ERT (nonusers). Bifrontal distance, the largest distance between frontal horns, and the size of ventricles were larger among current ERT users compared to past users or nonusers (P (trend) = .01), adjusted for all other covariates, but no dose-response relationship to current or past ERT use was found. Duration of estrogen use was not associated with any atrophy measure. Cortical atrophy measure, sulcal widening, or white matter disease did not differ significantly by ERT use or duration of use. Central measures of atrophy, bifrontal distance, and ventricular size were significantly associated with cognition as measured by MMSE. CONCLUSIONS: Current ERT users had much more clinically significant central atrophy than nonusers, but the implications remained unclear. PMID- 10811538 TI - Emotional well-being predicts subsequent functional independence and survival. AB - OBJECTIVE: To determine whether positive affect has an independent effect on functional status, mobility, and survival in an older Mexican American sample. DESIGN: A 2-year prospective cohort study. SETTING: Five Southwestern states: Texas, California, Arizona, New Mexico, and Colorado. PARTICIPANTS: A population based sample of 2282 Mexican Americans aged 65 to 99 who reported no functional limitations at baseline interview. MEASUREMENTS: In-home interviews in 1993-1994 and again in 1995-1996 assessed demographic variables, health conditions, activities of daily living, performance-based mobility, survival, and a rating of positive and negative affect. RESULTS: In multivariate analyses, there was a direct relationship between positive affect scores at baseline and mobility, functional status, and survival 2 years later, controlling for functional status, sociodemographic variables, major chronic conditions, body mass index (BMI), smoking status, drinking status, and negative affect at baseline. Subjects with high positive affect were half as likely (odds ratio (OR) = 0.48; 95% confidence interval (CI) 0.29, 0.93) to become disabled in activities of daily living (ADLs), two-thirds as likely (OR = 0.64; 95% CI 0.51, 0.79) to have a slow walking speed, and half as likely (OR 0.53; 95% CI 0.30, 0.93) to have died during the 2-year follow-up compared to those with lower positive affect scores. CONCLUSIONS: Our results support the concept that positive affect, or emotional well-being, is different from the absence of depression or negative affect. Positive affect seems to protect individuals against physical declines in old age. PMID- 10811539 TI - Distinction between preclinical Alzheimer's disease and depression. AB - OBJECTIVE: To assess the prevalence of depression in subjects with preclinical Alzheimer's disease (AD) and to investigate the possibility of differentiating subjects with preclinical AD and depression from subjects with depression-related cognitive impairment. DESIGN: A prospective, observational cohort study. SETTING: An outpatient memory clinic of a university-affiliated hospital. PARTICIPANTS: Nondemented subjects with cognitive impairment older than 55 years (n = 111) without neurological or somatic causes for the cognitive impairment. MEASUREMENTS: At baseline, data were collected on patient characteristics, the severity of depression, and cognitive functioning. The course of the cognitive impairment and the presence of dementia were assessed after 2 and 5 years. RESULTS: Twenty-five subjects had preclinical dementia with Alzheimer's type dementia at follow-up. Sixty percent of these subjects (n = 15) were depressed at baseline. Subjects with depression and preclinical AD had at baseline a poorer performance on the cognitive tasks and were older than the subjects with depression-related cognitive impairment. Logistic regression with backward step selection selected age and memory performance as the best predictors for Alzheimer's type dementia in the depressed subjects. The specificity of these predictors for the diagnosis of future Alzheimer's type dementia in depressed subjects was 94%, sensitivity was 90%, positive predictive value was 90%, and negative predictive value was 94%. CONCLUSIONS: Depression is common in preclinical AD. Depressed subjects with preclinical AD can be accurately differentiated from subjects with depression-related cognitive impairment by age and the severity of the memory impairment. Research that aims to investigate preclinical AD should not exclude a priori subjects with depression inasmuch as preclinical AD is often accompanied by depression. PMID- 10811540 TI - Improvement in quality-of-life measures and stimulation of weight gain after treatment with megestrol acetate oral suspension in geriatric cachexia: results of a double-blind, placebo-controlled study. AB - BACKGROUND: Weight loss among older patients is a severe problem, associated with an increased incidence of infections, decubiti, and death. Megestrol acetate (MA) causes weight gain in cachectic cancer and AIDS patients, but its effects in older cachectic patients are unknown. OBJECTIVE: To compare the effects of MA oral suspension (O.S.), 800 mg/day, versus placebo on weight in geriatric nursing home patients with weight loss or low body weight. DESIGN: Twelve-week, randomized, double-blind, placebo-controlled trial with a 13-week follow-up period. SETTING: Veterans Administration Medical Center (VMAC) nursing home. PATIENTS: Nursing home patients with weight loss of > or =5% of usual body weight over the past 3 months, or body weight 20% below their ideal body weight. INTERVENTIONS: Patients were randomly assigned to receive placebo or MA 800 mg/day for 12 weeks and were then followed for 13 weeks off treatment. MEASUREMENTS: Primary outcome was measured by weight and appetite change. Secondary outcome measures included sense of well-being, enjoyment of life, change in depression scale, laboratory nutrition parameters, energy intake counts, body composition, and adverse events. RESULTS: At 12 weeks there were no significant differences in weight gain between treatment groups, whereas MA treated patients reported significantly greater improvement in appetite, enjoyment of life, and well-being. Body composition was not statistically different between the two groups. At Week 25 (3 months after treatment), 61.9% of MA-treated patients had gained > or =1.82 kg (4 lbs) compared to 21.7% of placebo patients. CONCLUSIONS: In geriatric patients with weight loss or low body weight MA improves appetite and well-being after 12 weeks of treatment. During the 3 months of MA treatment, there was no statistically significant weight gain (> or =4 lbs). Three months after treatment, weight gain (> or =4 lbs) was significantly increased in MA-treated patients. PMID- 10811542 TI - The effects of physical activity on mortality in the Jerusalem 70-Year-Olds Longitudinal Study. AB - OBJECTIVE: To study the association between physical activity and mortality in older men and women. DESIGN: A community-based cohort study: the Jerusalem 70 Year-Olds Longitudinal Study. PARTICIPANTS: A systematically selected and representative sample of all residents of the western part of Jerusalem born in 1920-1921: 456 subjects, 25% of the total population. MEASUREMENTS: An extensive social and medical profile was developed by extensive interview and physical and ancillary examination. Medical diagnoses were established and subjects reported their level of regular physical activity. RESULTS: Unadjusted mortality at 6-year follow-up was significantly greater for subjects reporting no regular exercise than for those walking as little as 4 hours weekly (23.4% vs 9.9%, odds ratio (OR) = 2.77; 95% confidence interval (CI), 1.64-4.69). The significance of these benefits was demonstrated for males as well as for females (30.28% vs 12.14%, P < .001, 16.19% vs 6.86%, P = .036, respectively). Logistic regression analysis demonstrated the survival advantage to be independent of gender, smoking, subjective economic hardship, or preexisting medical conditions (hypertension; diabetes; coronary artery, cerebrovascular, renal, and respiratory diseases; anemia; and malignancy). Increased regularity of activity correlated with declining mortality. The odds ratios for mortality compared to the sedentary group were 0.73 (CI, 0.33-1.62) for those doing sports activity at least twice weekly, 0.41 (CI, 0.19-0.91) for those walking at least 4 hours weekly, 0.14 (CI, 0.04-0.50) for those exercising daily, and 0.40 (CI, 0.22-0.72) for all levels of physical activity combined. CONCLUSIONS: These results suggest that regular physical activity confers increased survival in the aged. It is proposed that older people be encouraged to engage in regular, moderate physical activity. PMID- 10811541 TI - Mobility difficulties and physical activity as predictors of mortality and loss of independence in the community-living older population. AB - OBJECTIVE: In older people, mobility impairments and physical inactivity are risk factors for further disability and death. We studied the interaction of physical activity and mobility impairment as a predictor of dependence and mortality. DESIGN: A population-based, prospective study. The data were collected in structured interviews in the year 1988 and 8 years later in the year 1996 as part of the Evergreen Project. PARTICIPANTS: Subjects were 1109 independently living, at baseline 65- to 84-year-old people in the city of Jyvaskyla, in central Finland. METHODS: Participants were ranked into four groups: (1) Intact mobility and physically active (Mobile-Active), (2) Intact mobility and sedentary (Mobile Sedentary), (3) Impaired mobility and physically active (Impaired-Active), and (4) Impaired mobility and sedentary (Impaired-Sedentary). The confounders adjusted for in the models included age, marital status, education, chronic conditions, smoking, and physical exercise earlier in life. RESULTS: In men and women, the relative risk of death was two times greater in Impaired-Active and three times greater in Impaired-Sedentary groups than the risk of death in Mobile Active groups. However, the risk of death did not differ between Mobile-Active and Mobile-Sedentary groups. The odds ratio for dependency (95% confidence interval) in Impaired-Sedentary men was 5.21 (1.44-18.70) and in Impaired Sedentary women was 2.92 (1.52-5.60) compared to Mobile-Active groups. The risk of dependence did not differ significantly between Mobile-Active, Mobile Sedentary, and Impaired-Active groups. CONCLUSIONS: Mobility impairments predicted mortality and dependence. However, among people with impaired mobility, physical activity was associated with lower risks, whereas the risk did not differ according to activity level among those with intact mobility. Despite of their overall greater risk, mobility-impaired people may be able to prevent further disability and mortality by physical exercise. PMID- 10811543 TI - Reasons given by older people for limitation or avoidance of leisure time physical activity. AB - OBJECTIVES: This study investigates reasons given by older people for limitation or avoidance of physical exercise. DESIGN: Community-based longitudinal study of free-living persons age 55 years or older in Sonoma, California. METHODS: Baseline standardized interviews were conducted (n = 2046) as part of a longitudinal study of aging and physical performance. Seventeen reasons for limitation or avoidance of physical activities were evaluated, with responses grouped as: no limitations, medical, non-medical, and combined medical and non medical reasons. Patterns of physical activity and medical conditions and direct measures of physical function were evaluated in relation to reasons for activity limitation by multinomial logistic regression. RESULTS: Women reported more limitations/avoidance of physical activity than men (81.0% vs 73.5%). Medical reasons increased with age. Reduced walking speed and depressive symptomatology were associated with a combination of medical and non-medical reasons in both sexes. Living arrangements were associated with non-medical reasons for women but were not for men. Non-medical reasons were associated with a 20 to 30% reduction in the number of subjects who reported physical activity at recommended levels. DISCUSSION: Non-medical factors are important determinants of activity limitation and should serve as targets for patient care providers and future interventions to improve participation in physical activity by older people. PMID- 10811544 TI - Community characteristics associated with elder abuse. AB - OBJECTIVES: To help define the relationship between elder abuse rates and counties' demographics, healthcare resources, and social service characteristics. DESIGN: County-level data from Iowa were analyzed to test the association between county characteristics and rates of elder abuse between 1984 and 1993 using univariate correlation analysis and stagewise linear regression. SETTING: Ninety nine counties in Iowa. PARTICIPANTS: Iowa residents aged 65 years and older. MEASUREMENTS: County-level population-adjusted numbers of abused elderly, abused children, children in poverty, high school dropouts, physicians and other healthcare providers, hospital beds, social workers and caseworkers in the Department of Human Services (DHS). RESULTS: Community characteristics that had a positive association with rates of reported or substantiated elder abuse at the P < .001 level were population density, children in poverty, and reported child abuse. Lower substantiated elder abuse rates were associated at P < .05 with higher community rates of high school dropouts, number of chiropractors, and number of nurse practitioners. After adjusting for number of DHS caseworkers and reported child abuse rates (a surrogate for workload) a district effect persists for substantiated elder abuse cases (P = .002). CONCLUSION: County demographics are risk factors for reported and substantiated elder abuse. The strongest risk factor for reported elder abuse was reported child abuse. The difference in districts may reflect differences in resources and/or differing characteristics of caseworkers who substantiate elder abuse. The risk factors may reflect conditions that influence the amount of elder abuse or the detection of existing elder abuse. PMID- 10811545 TI - Risk factors for nursing home placement in a population-based dementia cohort. AB - OBJECTIVE: To examine risk factors for nursing home placement in a population based dementia cohort. METHODS: The Mayo Clinic Medical Records linkage system was used to identify all patients with onset of dementia between 1980 and 1984. The patient group included 314 cases who met DSM-III-R criteria for dementia, including 220 cases who were community dwelling at onset. All dementia patients were followed until death. A control group included 323 patients who did not, at any point, meet DSM-III-R criterion for dementia. The groups were initially matched on age, gender, and year of initial registration. Demographic, medical, social, and functional predictors were examined as static and time-dependent risk factors for nursing home placement in the initial community-dwelling subgroups, using stepwise Cox regression modeling. RESULTS: Of the 314 dementia patients, 282 took residence in licensed skilled nursing homes for at least 6 weeks, suggestive of custodial care, at some point during the course of their illness. In the control group, 162 of the 323 people required nursing home placement. Within controls, the predictor variables of time to nursing home placement included initial age, being divorced, living in a townhome, apartment or assisted living apartment, change in Charlson comorbidity score, and change in amount of daily assistance required. Within the dementia sample, seven predictors were eventually determined to be associated with time to nursing home placement. These included total number of years of education, age at onset of dementia, being single, living in a retirement or supervised apartment at onset, change in Charlson comorbidity score, and a change in the amount of daily assistance required. CONCLUSIONS: Cumulative incidence of placement was 90% in the dementia cohort and 50% in the controls. Certain variables seem to impact time to nursing home placement in all older persons, whether they have dementia or not. Among these are age, living in assisted living settings, increasing comorbidity scores, and increasing need for functional assistance. Certain additional factors may have a specific impact in dementia. Among these is education, which seems to provide a protective effect. These predictors may be important covariates in clinical dementia studies that include time to nursing home placement as an outcome variable. PMID- 10811546 TI - Chair and bed rise performance in ADL-impaired congregate housing residents. AB - OBJECTIVES: To examine the ability of activity of daily living (ADL)-impaired older adults to successfully rise, and, when successful, the time taken to rise, from a bed and chair under varying rise task demands. SETTING: Seven congregate housing facilities SUBJECTS: Congregate housing residents (n = 116, mean age 82) who admitted to requiring assistance (such as from a person, equipment, or device) in performing at least one of the following mobility-related ADLs: transferring, walking, bathing, and toileting. METHODS: Subjects performed a series of bed and chair rise tasks where the rise task demand varied according to the head of bed (HOB) height, chair seat height, and use of hands. Bed rise tasks included supine to sit-to-edge, sit up in bed with hand use, and sit up in bed without hands, all performed from a bed where the HOB was adjusted to 0, 30, and 45 degrees elevations; roll to side-lying then rise (HOB 0 degrees); and supine to stand (HOB 0 degrees). Chair seat heights were adjusted according to the percent of the distance between the floor and the knee (% FK), and included rises (1) with hands and then without hands at 140, 120, 100, and 80% FK; (2) from a reclining (105 degrees at chair back) and tilting (seat tilted 10 degrees posteriorly) chair (100% FK); and (3) from a 80% FK seat height with a 4-inch cushion added, with and then without hands. Logistic regression for repeated measures was used to test for differences between tasks in the ability to rise. After log transformation of rise time, a linear effects model was used to compare rise time between tasks. RESULTS: The median total number of tasks successfully completed was 18 (range, 3-21). Nearly all subjects were able to rise from positions where the starting surface was elevated as long as hand use was unlimited. With the HOB at 30 or 45 degrees essentially all subjects could complete supine to sit-to-edge and sit up with hands. Essentially all subjects could rise from a seat height at 140, 120, and 100% FK as long as hand use was allowed. A small group (8-10%) of subjects was dependent upon hand use to perform the least challenging tasks, such as 140% FK without hands chair rise and 45 degrees sit up without hands. This dependency upon hand use increased significantly as the demand of the task increased, that is, as the HOB or seat height was lowered. Approximately three-quarters of the sample could not rise from a flat (0 degrees HOB elevation) bed or low (80% FK) chair when hand use was not allowed. Similar trends were seen in rise performance time, that is, performance times tended to increase as the HOB or chair seat elevation declined and as hand use was limited. Total self-reported ADL disability, compared to the single ADL transferring item, was a stronger predictor of rise ability and timed rise performance, particularly for chair rise tasks. CONCLUSIONS: Lowering HOB height and seat height increased bed and chair rise task difficulty, particularly when hand use was restricted. Restricting hand use in low HOB height or lowered seat height conditions may help to identify older adults with declining rise ability. Yet, many of those who could not rise under "without hands" conditions could rise under "with hands" conditions, suggesting that dependency on hand use may be a marker of progressive rise impairment but may not predict day-to-day natural milieu rise performance. Intertask differences in performance time may be statistically significant but are clinically small. Given the relationship between self-reported ADL disability and rise performance, impaired rise performance may be considered a marker for ADL disability. These bed and chair rise tasks can serve as outcomes for an intervention to improve bed and chair rise ability and might also be used in future studies to quantify improvements or declines in function over time, to refine physical therapy protocols, and to examine the effect of bed and chair design modifications on bed and chai PMID- 10811547 TI - Chiropractic care for patients aged 55 years and older: report from a practice based research program. AB - OBJECTIVE: To characterize patients aged 55 years and older and features of chiropractic care provided to them. DESIGN: Observational, practice-based research study. SETTING: Chiropractic offices in the United States and Canada, 1997-1998. PARTICIPANTS: Chiropractors in 96 practices in 32 states and two Canadian provinces collected data on 805 eligible patients aged 55 years and older during a 12-week study period. MEASUREMENTS: In addition to questionnaires on practice characteristics, patient demographics, chief complaints, and health habits, two standardized instruments were administered: for general health status, the Medical Outcomes Study 12-Item Short-Form Health Survey (SF-12); and for disability related to chronic pain, the Pain Disability Index (PDI). RESULTS: Of 805 study patients, 60.1% were women and' 94.7% were white. Overweight patients comprised 38.6% and obese 20.6% (n = 656) of the total; 9.7% of patients were hypertensive (n = 590). Smoking was reported by 12.7% and 50.2% reported regular exercise. The Physical Component Summary scores of the SF-12 seemed somewhat lower than population norms, whereas the Mental Component Summary scores differed very little from norms. Chief complaints were predominantly pain-related (72.3%), most commonly back pain (32.9%). The PDI mean baseline score for chronic patients was 16.3 (scale, 0-70), and 40.6% of study patients reported using at least one pain medication (prescription or nonprescription) more than three times per week. More than half of complaints (54.9%) had onsets more than 6 weeks before the baseline visit. For 66.6% of subjects, a chiropractor was the only provider for their current complaint. In addition to manipulation, most common features of care were recommendations on exercise (41.0%), heat or cold applications (40.8%), and food supplements (24.5%). At 4 weeks, 19.6% were discharged, 58.8% continued treatment, and 20.1% had discontinued care (self discharged). For these three groups, those with higher PDI mean baseline scores showed more change at 4 weeks. For patients who were discharged by the doctor, the proportion of reported pain medication use decreased 7.3% from baseline to 4 weeks, increased for patients who discontinued care, and remained about the same for those continuing care. CONCLUSIONS: Further investigation of the PDI and a decrease in pain medication use as outcome measures seems warranted. The descriptive information in this study may assist providers of care to older adults to better understand their patients' use of chiropractic care. PMID- 10811548 TI - A prospective study of the effect of fracture on measured physical performance: results from the MacArthur Study--MAC. AB - OBJECTIVE: To examine the impact of fractures on measured physical performance and to assess whether specific fractures have unique sequelae. SUBJECTS: 762 men and women, aged 70 to 79 at baseline, who were part of the MacArthur Study of Successful Aging. DESIGN: A longitudinal case-cohort: those with prevalent fractures at baseline were excluded; cases were persons with a medically diagnosed hip, arm, spine, or wrist fracture during the follow-up period (1988 1995). MEASURES: Eight physical performance tests: turning a circle, walking fast, chair stands, timed tap, tandem stand, grip strength, single leg stand, and balance (average of single leg and tandem stands) measured at baseline and follow up. ANALYSIS: Two fracture groups were defined: (1) those with incident wrist fractures (n = 7) and (2) those with a fracture of the hip, arm, or spine (combined fractures group, n = 16). Change in physical performance was analyzed using crude, age-adjusted, and multiply-adjusted ANCOVA models. RESULTS: The combined fracture group demonstrated statistically significant (P < .05) declines seven of eight of the performance tests compared with individuals without fractures. In contrast, individuals with wrist fractures did not experience a statistically significant decline in any performance measure compared with the no fracture group. CONCLUSIONS: Relative to those without fractures, individuals with a hip, arm, or clinical spinal fracture show similar global declines in physical performance, whereas those with wrist fracture demonstrate no physical performance decrements. PMID- 10811549 TI - Testosterone improves rehabilitation outcomes in ill older men. AB - OBJECTIVES: To determine whether testosterone supplementation improves rehabilitation outcomes in ill older men. DESIGN: A randomized, placebo controlled, double-blind study. SETTING: A Geriatric Evaluation and Management (GEM) unit based at a university- affiliated Veterans Affairs Medical Center. PARTICIPANTS: Fifteen men aged 65 to 90 years admitted to the GEM for rehabilitation. INTERVENTION: Subjects were randomized to receive weekly intramuscular injections with testosterone enanthate 100 mg or placebo. MEASUREMENTS: Task-specific performance using the Functional Independence Measure (FIM) and grip strength was measured at the onset of the study and at the time of discharge from the GEM. RESULTS: At baseline, FIM scores were similar between the placebo and the testosterone group (73.7 vs 70.7, P = .637), as was grip strength (49.7 vs 55.3 pounds, P = .555). At discharge from the GEM, testosterone-treated patients had improved FIM scores compared with baseline (93.6 vs 70.7; P = .012) and grip strength (68.7 vs 55.3 pounds; P = .033). In the placebo group there was no significant improvement of FIM scores compared with baseline (78.0 versus 73.7; P = .686) or of grip strength (48.9 vs 49.7 pounds; P = .686). CONCLUSIONS: Testosterone supplementation may improve rehabilitation outcomes in ill older men. PMID- 10811550 TI - Advance directive preferences among subpopulations of Asian nursing home residents in the Pacific Northwest. AB - OBJECTIVE: To study advance directives (code status) among subgroups of Asian nursing home residents. DESIGN: Cross-sectional design. PARTICIPANTS AND SETTING: A total of 423 residents of Asian descent (aged >55) from two ethnic nursing homes in Seattle, Washington. METHODS: Chart review was conducted on 423 residents (199 discharged between 1995 and 1998 and 244 current residents) to ascertain code status, age, gender, ethnicity, comorbidity (using the Charlson Index), and religion. RESULTS: Seventy percent of the residents were women, median age was 83 +/- 9, 43% were Chinese, 40% Japanese, and 17% other Asian (Korean, Filipino, Southeast Asian). The majority of the patients in any subgroup (72% overall) were 'no code'. In bivariate analysis, ethnicity, increased age, and comorbidity were correlated with no code status. In multivariable logistic regression, Japanese residents were more likely to be no code (OR 4.1 (95% CI, 3.1- 5.4)) controlling for age, comorbidity, gender, and religion. Chinese were more likely to be full code (OR 3.3 (95% CI, 2.6-4.2)). CONCLUSIONS: Code status differs significantly among Asian subgroups in these ethnic nursing homes. Whereas the majority of residents are no code, Japanese residents are more likely than Chinese or others to be no code. Higher age and comorbidity are also correlated with no code status. PMID- 10811551 TI - Interrater reliability of the Clinical Dementia Rating in a multicenter trial. AB - OBJECTIVE: To test the interrater reliability of the Clinical Dementia Rating (CDR) in a multicenter clinical trial. DESIGN: Observational study. SETTING: Training session for a multicenter trial of milameline, a direct muscarinic agonist, in the treatment of Alzheimer's disease. PARTICIPANTS: Twenty-four raters (physicians and nurses) familiar with drug trials and expert in the care of patients with Alzheimer's disease. METHODS: Independent scoring of the CDR using four videotaped CDR interviews. OUTCOME MEASURE: Interrater reliability, as tested by the Kappa statistic RESULTS: The overall interrater reliability was 0.62. Within the CDR domains, the global kappas ranged from 0.33 +/- 0.06 to 0.88 +/- 0.06. CONCLUSIONS: The data support moderate to high overall interrater reliability but show important difficulties in the reliable assessment of early dementia. PMID- 10811552 TI - Update on electroconvulsive therapy (ECT) in older adults. AB - Electroconvulsive therapy (ECT) is used increasingly in the older adult population for major depression, particularly when depression is not responsive to medications, when antidepressants are not tolerated due to side effects, or when depression is accompanied by life-threatening complications such as severe weight loss or catatonia where a rapid definitive response is required. ECT is considered a low-risk procedure that can be successfully done in medically ill older adults, but it is associated with a brief period of increased blood pressure and pulse leading to increased myocardial oxygen demand. ECT may cause delirium, particularly in the cognitively impaired older. As successful management of older patients undergoing a course of ECT often involves geriatricians and other medical practitioners, this review provides an update on the indications for ECT, how it is done, the common complications seen after the procedure, and its efficacy. Finally, specific recommendations for management are made. PMID- 10811553 TI - Stair negotiation in older people: a review. AB - Locomotion on stairs is among the most challenging and hazardous activities of daily living for older individuals. This is evidenced by the reports that stair falls account for more than 10% of fatal fall accidents. The demands that stairs place on the musculoskeletal and cardiovascular systems are compounded by the need for input from the somatosensory, visual, and vestibular systems at various stages in the task. Many of these collaborating systems deteriorate with aging, thus increasing the difficulty and risk of failure in a task that inherently involves exposure to significant danger. The task itself varies in its degree of challenge depending on many structural and environmental factors that are outside the control of the stair user. This review explores existing literature in the area of stair negotiation by older persons in an effort to define the key factors associated with difficulty and safety on stairs and to interpret these findings in the light of possible interventions that could increase stair safety. Further research is needed to ensure that current recommendations for stair design are suitable for older stair users. A Glossary of Terms is also provided. PMID- 10811554 TI - Implementation of an interdisciplinary Behavior Management Program. AB - OBJECTIVE: To describe the establishment and utilization of an interdisciplinary Behavior Management Program in a Transitional Care Unit as a new model of Geriatrics Practice. DESIGN: Descriptive Study. SETTING: A 50-bed Transitional Care Unit in a Veterans Affairs Medical Center in Syracuse, New York. PARTICIPANTS: All residents admitted to the Transitional Care Unit were observed for a 6-month period under the Behavior Management Program. PROGRAM STRUCTURE: The Behavior Management Program involved an interdisciplinary team approach to the management of behavioral disturbances. This included development of a Behavior Team and a comprehensive Behavior Observation Record, which readily assisted caregivers in determining and monitoring target behaviors that required modification. Both nonpharmacological and pharmacological interventions were employed to assist in the modification of behavioral disturbances. RESULTS: The Behavior Management Program enhanced the overall care of Transitional Care Unit residents with behavioral disturbances. The more appropriate use of medications for specific target behaviors led to a reduction in behavioral disturbance episodes and increased the earlier diagnosis and treatment of depression. The interdisciplinary nature of the Behavior Management Program promoted educational efforts and camaraderie among staff and created a stronger liaison between the Transitional Care Unit and the Psychiatry Service. In addition, several management improvements were identified as a result of the implementation of the Behavior Management Program. CONCLUSIONS: An interdisciplinary Behavior Management Program can prove to be a valuable asset in the management of behaviors for people with dementia, psychiatric illness, and other medical illnesses. More effective use of nonpharmacological and psychopharmacological interventions enhances the quality of life in a frail older population. This approach has strong potential for successful use in a variety of clinical settings by providers and caregivers alike. PMID- 10811555 TI - Estrogen replacement therapy, cognition, and cerebral changes by MRI in the cardiovascular health study. PMID- 10811556 TI - A happy person, a healthy person? PMID- 10811557 TI - A case of mistaken identity: pseudodementia is really predementia. PMID- 10811558 TI - CPR in nursing homes. PMID- 10811559 TI - Considering CPR policy. PMID- 10811560 TI - Nurse case management of frail older people. PMID- 10811561 TI - Drug interactions and donepezil. PMID- 10811562 TI - Calculating bone-lead measurement variance. AB - The technique of (109)Cd-based X-ray fluorescence (XRF) measurements of lead in bone is well established. A paper by some XRF researchers [Gordon CL, et al. The Reproducibility of (109)Cd-based X-ray Fluorescence Measurements of Bone Lead. Environ Health Perspect 102:690-694 (1994)] presented the currently practiced method for calculating the variance of an in vivo measurement once a calibration line has been established. This paper corrects typographical errors in the method published by those authors; presents a crude estimate of the measurement error that can be acquired without computational peak fitting programs; and draws attention to the measurement error attributable to covariance, an important feature in the construct of the currently accepted method that is flawed under certain circumstances. PMID- 10811564 TI - Groundwater arsenic contamination in Bangladesh and West Bengal, India. AB - Nine districts in West Bengal, India, and 42 districts in Bangladesh have arsenic levels in groundwater above the World Health Organization maximum permissible limit of 50 microg/L. The area and population of the 42 districts in Bangladesh and the 9 districts in West Bengal are 92,106 km(2) and 79.9 million and 38,865 km(2) and 42.7 million, respectively. In our preliminary study, we have identified 985 arsenic-affected villages in 69 police stations/blocks of nine arsenic-affected districts in West Bengal. In Bangladesh, we have identified 492 affected villages in 141 police stations/blocks of 42 affected districts. To date, we have collected 10,991 water samples from 42 arsenic-affected districts in Bangladesh for analysis, 58,166 water samples from nine arsenic-affected districts in West Bengal. Of the water samples that we analyzed, 59 and 34%, respectively, contained arsenic levels above 50 microg/L. Thousands of hair, nail, and urine samples from people living in arsenic-affected villages have been analyzed to date; Bangladesh and West Bengal, 93 and 77% samples, on an average, contained arsenic above the normal/toxic level. We surveyed 27 of 42 districts in Bangladesh for arsenic patients; we identified patients with arsenical skin lesions in 25 districts. In West Bengal, we identified patients with lesions in seven of nine districts. We examined people from the affected villages at random for arsenical dermatologic features (11,180 and 29,035 from Bangladesh and West Bengal, respectively); 24.47 and 15.02% of those examined, respectively, had skin lesions. After 10 years of study in West Bengal and 5 in Bangladesh, we feel that we have seen only the tip of iceberg. PMID- 10811563 TI - The PBDEs: an emerging environmental challenge and another reason for breast-milk monitoring programs. AB - Levels of the polybrominated diphenyl ethers (PBDEs), a class of widely used flame retardants, appear to be rising rapidly in human tissues, as evidenced by studies of human breast milk. The case of the PBDEs illustrates the value of breast-milk monitoring programs in identifying important emerging pollutants, and highlights why such monitoring programs are needed in the United States. A review of the use, occurrence, and toxicity of PBDEs indicates many parallels between some PBDEs, PCBs, and other polyhalogenated persistent organic pollutants, and suggests that the PBDEs may be a significant environmental challenge in the future. PMID- 10811565 TI - Chromium(III)-induced 8-hydroxydeoxyguanosine in DNA and its reduction by antioxidants: comparative effects of melatonin, ascorbate, and vitamin E. AB - Chromium compounds are well documented carcinogens. Cr(III) is more reactive than Cr(VI) toward DNA under in vitro conditions. In the present study, we investigated the ability of Cr(III) to induce oxidative DNA damage by examining the formation of 8-hydroxydeoxyguanosine (8-OH-dG) in calf thymus DNA incubated with CrCl(3) plus H(2)O(2). We measured 8-OH-dG using HPLC with electrochemical detection. In the presence of H(2)O(2), we observed that Cr(III)-induced formation of 8-OH-dG in isolated DNA was dose and time dependent. Melatonin, ascorbate, and vitamin E (Trolox), all of which are free radical scavengers, markedly inhibited the formation of 8-OH-dG in a concentration-dependent manner. The concentration that reduced DNA damage by 50% was 0.51, 30.4, and 36.2 microM for melatonin, ascorbate, and Trolox, respectively. The results show that melatonin is 60- and 70-fold more effective than ascorbate or vitamin E, respectively, in reducing oxidative DNA damage in this in vitro model. These findings also are consistent with the conclusion that the carcinogenic mechanism of Cr(III) is possibly due to Cr(III)-mediated Fenton-type reactions and that melatonin's highly protective effects against Cr(III) relate, at least in part, to its direct hydroxyl radical scavenging ability. PMID- 10811566 TI - Assaying estrogenicity by quantitating the expression levels of endogenous estrogen-regulated genes. AB - Scientific evidence suggests that humans and wildlife species may experience adverse health consequences from exposure to environmental chemicals that interact with the endocrine system. Reliable short-term assays are needed to identify hormone-disrupting chemicals. In this study we demonstrate that the estrogenic activity of a chemical can be evaluated by assaying induction or repression of endogenous estrogen-regulated "marker genes" in human breast cancer MCF-7 cells. We included four marker genes in the assay--pS2, transforming growth factor beta3 (TGFbeta3), monoamine oxidase A, and [alpha]1-antichymotrypsin--and we evaluated estrogenic activity for 17beta-estradiol (E(2)), diethylstilbestrol, [alpha]-zearalanol, nonylphenol, genistein, methoxychlor, endosulphan, o,p-DDE, bisphenol A, dibutylphthalate, 4-hydroxy tamoxifen, and ICI 182.780. All four marker genes responded strongly to the three high-potency estrogens (E(2), diethylstilbestrol, and [alpha]-zearalanol), whereas the potency of the other chemicals was 10(3)- to 10(6)-fold lower than that of E(2). There were some marker gene-dependent differences in the relative potencies of the tested chemicals. TGFbeta3 was equally sensitive to the three high-potency estrogens, whereas the sensitivity to [alpha]-zearalanol was approximately 10-fold lower than the sensitivity to E(2) and diethylstilbestrol when assayed with the other three marker genes. The potency of nonylphenol was equal to that of genistein when assayed with pS2 and TGFbeta3, but 10- to 100-fold higher/lower with monoamine oxidase A and [alpha]1-antichymotrypsin, respectively. The results are in agreement with results obtained by other methods and suggest that an assay based on endogenous gene expression may offer an attractive alternative to other E-SCREEN methods. PMID- 10811567 TI - Metallothionein isoform 3 as a potential biomarker for human bladder cancer. AB - The goal of the present study was to determine if the expression of metallothionein isoform 3 (MT-3) might serve as a biomarker for human bladder cancer. To accomplish this goal, we defined the localization and expression of MT 3 protein and mRNA using fresh and archival biopsy specimens obtained from patients undergoing differential diagnosis for a variety of bladder disorders. We used immunohistochemistry, immunoblot, and RT-PCR analysis to define the localization and expression of MT-3 protein and mRNA. Immunohistochemical analysis disclosed no immunoreactivity for MT-3 in normal bladder cells. The absence of MT-3 expression in the normal bladder was further confirmed by demonstrating that MT-3 mRNA could not be detected using reverse transcriptase polymerase chain reaction (RT-PCR) or MT-3 protein using immunoblot. Immunohistochemistry also disclosed no immunoreactivity for MT-3 in archival biopsy specimens from patients with interstitial cystitis and related disorders. Immunohistochemical analysis demonstrated that MT-3 was expressed in carcinoma in situ (CIS), high-grade bladder cancer, low-grade bladder cancer, and dysplastic lesions. MT-3 immunostaining was intense in both CIS and high-grade bladder cancer, and low to moderate in low-grade bladder cancer and dysplastic lesions. We determined MT-3 mRNA expression in a subset of these bladder cancer specimens; expression was elevated as compared to that of the housekeeping gene, ss-actin. The cDNA from the RT-PCR reaction primed for MT-3 contained a FokI restriction site, a site unique for MT-3 as compared to other MT family members. In conclusion, this study demonstrates that MT-3 is up-regulated in human bladder cancer and that this up-regulation increases with increasing tumor grade. The finding that MT-3 expression is minimal in normal bladder suggests that MT-3 might be developed into an effective biomarker for bladder cancer. PMID- 10811568 TI - Exposure measurement error in time-series studies of air pollution: concepts and consequences. AB - Misclassification of exposure is a well-recognized inherent limitation of epidemiologic studies of disease and the environment. For many agents of interest, exposures take place over time and in multiple locations; accurately estimating the relevant exposures for an individual participant in epidemiologic studies is often daunting, particularly within the limits set by feasibility, participant burden, and cost. Researchers have taken steps to deal with the consequences of measurement error by limiting the degree of error through a study's design, estimating the degree of error using a nested validation study, and by adjusting for measurement error in statistical analyses. In this paper, we address measurement error in observational studies of air pollution and health. Because measurement error may have substantial implications for interpreting epidemiologic studies on air pollution, particularly the time-series analyses, we developed a systematic conceptual formulation of the problem of measurement error in epidemiologic studies of air pollution and then considered the consequences within this formulation. When possible, we used available relevant data to make simple estimates of measurement error effects. This paper provides an overview of measurement errors in linear regression, distinguishing two extremes of a continuum-Berkson from classical type errors, and the univariate from the multivariate predictor case. We then propose one conceptual framework for the evaluation of measurement errors in the log-linear regression used for time series studies of particulate air pollution and mortality and identify three main components of error. We present new simple analyses of data on exposures of particulate matter < 10 microm in aerodynamic diameter from the Particle Total Exposure Assessment Methodology Study. Finally, we summarize open questions regarding measurement error and suggest the kind of additional data necessary to address them. PMID- 10811569 TI - Air pollution and daily hospital admissions in metropolitan Los Angeles. AB - We used daily time-series analysis to evaluate associations between ambient carbon monoxide, nitrogen dioxide, particulate matter [less than and equal to] 10 microm in aerodynamic diameter (PM(10)), or ozone concentrations, and hospital admissions for cardiopulmonary illnesses in metropolitan Los Angeles during 1992 1995. We performed Poisson regressions for the entire patient population and for subgroups defined by season, region, or personal characteristics, allowing for effects of temporal variation, weather, and autocorrelation. CO showed the most consistently significant (p<0.05) relationships to cardiovascular admissions. A wintertime 25th-75th percentile increase in CO (1.1-2.2 ppm) predicted an increase of 4% in cardiovascular admissions. NO(2), and, to a lesser extent, PM(10) tracked CO and showed similar associations with cardiovascular disease, but O(3) was negatively or nonsignificantly associated. No significant demographic differences were found, although increased cardiovascular effects were suggested in diabetics, in whites and blacks (relative to Hispanics and Asians), and in persons older than 65 years of age. Pulmonary disease admissions associated more with NO(2) and PM(10) than with CO. Pulmonary effects were generally smaller than cardiovascular effects and were more sensitive to the choice of model. We conclude that in Los Angeles, atmospheric stagnation with high primary (CO/NO(2)/PM(10)) pollution, most common in autumn/winter, increases the risk of hospitalization for cardiopulmonary illness. Summer photochemical pollution (high O(3)) apparently presents less risk. PMID- 10811570 TI - Assessing potential health risks from microcystin toxins in blue-green algae dietary supplements. AB - The presence of blue-green algae (BGA) toxins in surface waters used for drinking water sources and recreation is receiving increasing attention around the world as a public health concern. However, potential risks from exposure to these toxins in contaminated health food products that contain BGA have been largely ignored. BGA products are commonly consumed in the United States, Canada, and Europe for their putative beneficial effects, including increased energy and elevated mood. Many of these products contain Aphanizomenon flos-aquae, a BGA that is harvested from Upper Klamath Lake (UKL) in southern Oregon, where the growth of a toxic BGA, Microcystis aeruginosa, is a regular occurrence. M. aeruginosa produces compounds called microcystins, which are potent hepatotoxins and probable tumor promoters. Because M. aeruginosa coexists with A. flos-aquae, it can be collected inadvertently during the harvesting process, resulting in microcystin contamination of BGA products. In fall 1996, the Oregon Health Division learned that UKL was experiencing an extensive M. aeruginosa bloom, and an advisory was issued recommending against water contact. The advisory prompted calls from consumers of BGA products, who expressed concern about possible contamination of these products with microcystins. In response, the Oregon Health Division and the Oregon Department of Agriculture established a regulatory limit of 1 microg/g for microcystins in BGA-containing products and tested BGA products for the presence of microcystins. Microcystins were detected in 85 of 87 samples tested, with 63 samples (72%) containing concentrations > 1 microg/g. HPLC and ELISA tentatively identified microcystin-LR, the most toxic microcystin variant, as the predominant congener. PMID- 10811572 TI - Quantitative polymerase chain reaction for transforming growth factor-beta applied to a field study of fish health in Chesapeake Bay tributaries. AB - Fish morbidity and mortality events in Chesapeake Bay tributaries have aroused concern over the health of this important aquatic ecosystem. We applied a recently described method for quantifying mRNA of an immunosuppressive cytokine, transforming growth factor-beta (TGF-beta), by reverse transcription quantitative competitive polymerase chain reaction to a field study of fish health in the Chesapeake Basin, and compared the results to those of a traditional cellular immunoassay macrophage bactericidal activity. We selected the white perch (Morone americana) as the sentinel fish species because of its abundance at all of the collection sites. White perch were sampled from Chesapeake Bay tributaries in June, August, and October 1998. Splenic mononuclear cell TGF-beta mRNA levels increased and anterior kidney macrophage bactericidal activity decreased, particularly in eastern shore tributaries, from June to August and October. The results of the two assays correlated inversely (Kendall's [Tau] b = -0.600; p = 0.0102). The results indicated both temporal and spatial modulation of white perch immune systems in the Chesapeake Basin, and demonstrated the utility of quantitative PCR for TGF-beta as a molecular biomarker for field assessment of teleost fish immune status. PMID- 10811571 TI - Concentrations of organochlorines related to titers to Epstein-Barr virus early antigen IgG as risk factors for hairy cell leukemia. AB - Hairy cell leukemia (HCL) is a rare chronic B-cell malignancy that, according to modern classifications, is a subgroup of non-Hodgkin lymphomas (NHLs). A rapid increase in incidence of NHL has been reported in many countries. The reasons for this increase are largely unknown, but exposure to organochlorines has been suggested as a risk factor. Epstein-Barr virus is a human herpesvirus that has been associated with certain subgroups of NHL. In this study, we measured lipid adjusted blood concentrations (in nanogram per gram) of 36 congeners of polychlorinated biphenyls (PCBs), p, p'-dichlorodiphenyldichloroethylene (p,p' DDE), hexachlorobenzene (HCB), and four subgroups of chlordanes (trans-nonachlor, cis-nonachlor, MC6, and oxychlordane) in incident cases of HCL and controls from the general population. We obtained results on organochlorines and antibodies for 54 cases and 54 controls. Titers of antibodies to the Epstein-Barr early antigen and Epstein-Barr nuclear antigen, measured as P107, were correlated to concentrations of organochlorines to evaluate the possibility of an interaction between these factors in the pathogenesis of HCL. We found no significant difference in lipid-adjusted blood concentrations of total PCBs, p,p'-DDE, HCB, or the sum of the chlordanes between cases and controls. Titers of antibodies to Epstein-Barr early antigen IgG [Greater and equal to] 40 were correlated to an increased risk for HCL. This risk was further increased in those with a level above the median value of p,p'-DDE, HCB, or the sum of the chlordanes, suggesting an interaction between Epstein-Barr virus and a higher concentration of these chemicals. We also found increased risk for the sum of immunotoxic PCB group. PMID- 10811573 TI - A pilot study examining changes in dust lead loading on walls and ceilings after lead hazard control interventions. AB - The U.S. Department of Housing and Urban Development (HUD) guidelines on lead hazard control instruct contractors to clean floors, windows, walls, ceilings, and other horizontal surfaces to remove lead-contaminated dust and debris after lead interventions are conducted. This dust removal activity adds costs to each project. The need to clean floors and windows is well documented in the HUD guidelines. However, there is substantially less documentation to support the recommendation to clean walls and ceilings. We examined whether it is necessary to clean walls and ceilings after lead hazard control (LHC) interventions by comparing dust lead loadings measured on these surfaces before an LHC intervention to dust lead loadings after the intervention. Twenty-two dwelling units undergoing substantial LHC measures consistent with the HUD guidelines were enrolled in the study. There was a significant increase in dust lead loading on walls and ceilings between the pre- and postintervention. The change in wall dust lead loading was substantial and created potentially harmful lead exposures. Although statistically significant, the change in ceiling dust lead loading was minimal and the postintervention dust lead loadings were far below the existing federal floor dust lead clearance standard. These results strongly support the recommendations in the HUD guidelines to clean walls after LHC interventions and do not provide sufficient justification to alter the current recommendation to clean ceilings after lead work. PMID- 10811574 TI - Does the risk of childhood diabetes mellitus require revision of the guideline values for nitrate in drinking water? AB - In recent years, several studies have addressed a possible relationship between nitrate exposure and childhood type 1 insulin-dependent diabetes mellitus. The present ecologic study describes a possible relation between the incidence of type 1 diabetes and nitrate levels in drinking water in The Netherlands, and evaluates whether the World Health Organization and the European Commission standard for nitrate in drinking water (50 mg/L) is adequate to prevent risk of this disease. During 1993-1995 in The Netherlands, 1,104 cases of type 1 diabetes were diagnosed in children 0-14 years of age. We were able to use 1,064 of these cases in a total of 2,829,020 children in this analysis. We classified mean nitrate levels in drinking water in 3,932 postal code areas in The Netherlands in 1991-1995 into two exposure categories. One category was based on equal numbers of children exposed to different nitrate levels (0.25-2.08, 2.10-6.42, and 6.44 41.19 mg/L nitrate); the other was based on cut-off values of 10 and 25 mg/L nitrate. We determined standardized incidence ratios (SIRs) for type 1 diabetes in subgroups of the 2,829,020 children with respect to both nitrate exposure categories, sex, and age and as compared in univariate analysis using the chi square test for trend. We compared the incidence rate ratios (IRRs) by multivariate analysis in a Poisson regression model. We found an effect of increasing age of the children on incidence of type 1 diabetes, but we did not find an effect of sex or of nitrate concentration in drinking water using the two exposure categories. For nitrate levels > 25 mg/L, an increased SIR and an increased IRR of 1.46 were observed; however, this increase was not statistically significant, probably because of the small number of cases (15 of 1,064). We concluded that there is no convincing evidence that nitrate in drinking water at current exposure levels is a risk factor for childhood type 1 diabetes mellitus in The Netherlands, although a threshold value > 25 mg/L for the occurrence of this disease can not be excluded. PMID- 10811575 TI - Is the incidence of hypospadias increasing? Analysis of Finnish hospital discharge data 1970-1994. AB - Reports suggesting an increasing incidence of male genitourinary anomalies such as hypospadias, possibly related to environmental factors such as environmental estrogen-like compounds, have recently received considerable publicity. These reports are based on birth defects registry data, and there may be variation in the completeness of the registries used. We analyzed temporal trends in the prevalence of hypospadias in Finland to assess the previously reported low overall prevalence and to detect any possible increasing tendencies during the past decade. We identified all patients who were surgically treated for hypospadias before the age of 9 years among boys born 1970-1986 in the national hospital discharge registry. We calculated the cumulative prevalence by dividing the number of patients by the number of male births, and we used Poisson regression analysis. Out of 549,176 boys born in Finland in 1970-1986, 1,543 were treated for hypospadias by the age of 8 years (28.1 surgically treated patients per 10,000 male live births; 95% confidence interval, 26.7-29.5). The prevalence of hypospadias in Finland remained constant throughout the study period and appears to have been approximately three times higher than previously reported. Changes in completeness of registration may account for a substantial proportion of the reported increases in the prevalence of hypospadias in Finland and possibly also elsewhere. PMID- 10811576 TI - Scientific issues in relation to lakewide management plans: linking science and policy. PMID- 10811577 TI - The genomic era: a crucial role for the public health sciences. PMID- 10811578 TI - Endocrine disruption: lessons learned. PMID- 10811579 TI - Radio-free bacteria. PMID- 10811580 TI - Accounting for lost acreage. PMID- 10811581 TI - Plague linked to precipitation. PMID- 10811582 TI - Lights, chemicals, action at new lab for phototoxicology. PMID- 10811583 TI - Trampling paradise: dream vacation--environmental nightmare? PMID- 10811584 TI - Global business: operating in the green. PMID- 10811585 TI - May 16, 2000 PMID- 10811586 TI - Thrombolytic therapy for patients with myocardial infarction who are older than 75 years. Do the risks outweigh the benefits? PMID- 10811587 TI - Safety of intracoronary gamma-radiation on uninjured reference segments during the first 6 months after treatment of in-stent restenosis: a serial intravascular ultrasound study. AB - BACKGROUND: The effects of endovascular irradiation on uninjured reference segments during the treatment of in-stent restenosis are unknown. METHODS AND RESULTS: In the Washington Radiation for In-Stent restenosis Trial (WRIST), patients with in-stent restenosis were first treated with conventional catheter based techniques and then randomized (blinded) to receive either gamma irradiation ((192)Ir) or a placebo (dummy seeds). We identified all patients in whom the active (n=19) or dummy seeds (n=19) extended >10 mm proximal and distal to the in-stent restenosis lesion. Serial (postirradiation and follow-up) external elastic membrane (EEM), lumen, and plaque and media (EEM-lumen) areas were measured (using intravascular ultrasound) every 1 mm over 5-mm-long reference segments that were 6 to 10 mm proximal and distal to the in-stent restenosis lesion. During follow-up, a similar small increase occurred in the plaque and media area in the proximal and distal reference segments in both (192)Ir and placebo patients. However, in the (192)Ir patients, an increase in both proximal and distal EEM area occurred; as a result, no change in lumen area occurred. Conversely, in the placebo patients, the proximal reference EEM area decreased, and no change occurred in the distal reference EEM area; this contributed to a decrease in lumen area. CONCLUSIONS: There was no evidence of a deleterious effect of gamma-irradiation on angiographically normal uninjured reference segments in the first 6 months after the treatment of in-stent restenosis. PMID- 10811588 TI - Prediction of 1-year survival after thrombolysis for acute myocardial infarction in the global utilization of streptokinase and TPA for occluded coronary arteries trial. AB - BACKGROUND: When a patient survives thrombolysis for acute myocardial infarction, little information from large studies exists from which to estimate prognosis during follow-up visits. METHODS AND RESULTS: Baseline, in-hospital, and later survival data were collected from 41 021 patients enrolled in Global Utilization of Streptokinase and TPA for Occluded Coronary Arteries, a randomized trial of 4 thrombolytic-heparin regimens with standard aspirin and beta-blockade. Cox proportional hazards models were developed to predict 1-year survival in 30-day survivors (n=37 869) from baseline clinical and ECG factors and in-hospital factors; a combined model then was developed (C-index 0.800). The model was simplified into a nomogram to predict individual outcomes (C-index 0.754). Factors reflecting demographics (advanced age, lighter weight), larger infarctions (higher Killip class, lower blood pressure, faster heart rate, longer QRS duration), cardiac risk (smoking, hypertension, prior cerebrovascular disease), and arrhythmia were important predictors of death between 30 days and 1 year. Black race was associated with a substantial increase in risk after considering other factors. Revascularization was associated with reduced risk between 30 days and 1 year. CONCLUSIONS: When evaluating a patient who has survived acute infarction treated with thrombolysis, clinicians can estimate the likelihood of survival from factors easily measured during admission. Although many risk factors clearly relate to age, left ventricular dysfunction, or clinical instability, black race is an unexplained risk factor requiring further examination. PMID- 10811589 TI - Lack of benefit for intravenous thrombolysis in patients with myocardial infarction who are older than 75 years. AB - BACKGROUND: The benefit of intravenous thrombolytic therapy in elderly patients with myocardial infarction is uncertain. There are no randomized trials of thrombolytic efficacy or observational studies of clinical effectiveness that focus specifically on the elderly. METHODS AND RESULTS: To determine whether thrombolytic therapy for elderly patients is associated with a survival advantage in a large observational database, we conducted a retrospective cohort study of 7864 Medicare fee-for-service patients aged 65 to 86 years with the primary discharge diagnosis of acute myocardial infarction who were admitted with clinical and ECG indications for thrombolytic therapy and no absolute contraindications. The study included all US acute care nongovernment hospitals without on-site angioplasty capability. Using proportional-hazards methods, we found that in a comprehensive multivariate model, there was a significant interaction (P<0.001) between age and the effect of thrombolytic therapy on 30 day mortality rates. For patients 65 to 75 years old, thrombolytic therapy was associated with a survival benefit, consistent with randomized trials. Among patients aged 76 to 86 years, thrombolytic therapy was associated with a survival disadvantage, with a 30-day mortality hazard ratio of 1.38 (95% CI 1. 12 to 1.71, P=0.003). For these patients, there was no benefit from thrombolytic therapy in any clinical subgroup. CONCLUSIONS: In nationwide clinical practice, thrombolytic therapy for patients >75 years old is unlikely to confer survival benefit and may have a significant survival disadvantage. Reperfusion research that is focused on elderly patients is urgently needed. PMID- 10811590 TI - Circulating adhesion molecules in humans: role of hyperglycemia and hyperinsulinemia. AB - BACKGROUND: We assessed the role of glucose and insulin in the regulation of circulating levels of soluble intercellular adhesion molecule-1 (sICAM-1) and vascular adhesion molecule-1 (sVCAM-1) in normal subjects and in patients with type 2 diabetes. METHODS AND RESULTS: Plasma glucose concentrations were acutely raised in 10 normal subjects and 10 newly diagnosed, complication-free type 2 diabetic patients and maintained at 15 mmol/L for 2 hours. In normal subjects, plasma sICAM-1, but not sVCAM-1, levels rose significantly (P<0.01) at 1 hour and returned to basal values at 2 hours. In another study, octreotide was infused during the hyperglycemic clamp to block the release of endogenous insulin; this prevented the late fall of plasma sICAM-l levels observed in under control clamp conditions. The diabetic patients had plasma sICAM-1 levels significantly higher (P<0.01) than those of the control subjects; plasma sVCAM-1 levels were similar. Both sICAM-l and sVCAM-1 concentrations did not change significantly during the control hyperglycemic clamp; however, octreotide infusion increased plasma sICAM 1 levels, which remained significantly (P<0.05) above baseline during the whole clamp. In an additional 10 type 2 diabetic patients, overnight euglycemia (plasma glucose 5.5 mmol/L) obtained with the aid of an artificial pancreas or supplementation with l-arginine (10 g PO for 30 days), the natural precursor of NO, normalized the increased plasma sICAM-1 levels. CONCLUSIONS: Acute hyperglycemia increases circulating sICAM-1 levels in normal subjects, whereas the correction of hyperglycemia with insulin or l-arginine supplementation restored to normal levels the increased plasma sICAM-1 levels of type 2 diabetic patients. PMID- 10811591 TI - Smoking is associated with reduced serum paraoxonase activity and concentration in patients with coronary artery disease. AB - BACKGROUND: Paraoxonase is an HDL-associated enzyme that protects lipoproteins from oxidative modifications. Smoking is a major cardiovascular risk factor that promotes lipid peroxidation. Cigarette smoke has been shown in vitro to inhibit paraoxonase. The present study examined the hypothesis that smoking is associated with modulated serum activities and concentrations of paraoxonase. METHODS AND RESULTS: Coronary artery disease was assessed with the use of coronary arteriography in participants recruited from a hospital cardiology division. Medical and lifestyle data were obtained, and a fasting blood sample was provided. Three smoking categories were established (never, ex-smokers, and current smokers), and serum paraoxonase variables were compared among them. The activities and concentrations of paraoxonase were significantly lower in current than in never smokers. Ex-smokers had values comparable to those of never smokers. Ex-smokers who had recently stopped (<3 months) had activities and concentrations comparable to those of current smokers; values returned to the levels of never smokers within 2 years of cessation of smoking. Smoking status was an independent determinant of paraoxonase activity and concentration in multivariate analysis. Finally, lower paraoxonase was associated with more severe coronary disease and a reduced capacity to protect LDL from oxidation. CONCLUSIONS: Smoking is independently associated with significant decreases in serum paraoxonase activities and concentrations, which normalize within a relatively short time of cessation. Lower serum paraoxonase is linked to more severe coronary artery disease and a lower antioxidant capacity. The data are consistent with the hypothesis that smoking modifies serum paraoxonase such that there is an increased risk of coronary artery disease due to a diminished capacity to protect lipoproteins from oxidative stress. PMID- 10811592 TI - Mechanisms responsible for endothelial dysfunction associated with acute estrogen deprivation in normotensive women. AB - BACKGROUND: The goal of this study was to evaluate whether endothelial dysfunction associated with acute estrogen deprivation is caused by an alteration in the L-arginine-nitric oxide (NO) pathway and oxidative stress. Methods and Results-In 26 healthy women (age, 45.7+/-5.4 years) and 18 fertile women with leiomyoma (age, 44.5+/-5.1 years), we studied forearm blood flow (strain-gauge plethysmography) changes induced by intrabrachial acetylcholine (0. 15, 0.45, 1.5, 4.5, or 15 microgram. 100 mL(-1). min(-1)) or sodium nitroprusside (1, 2, or 4 microgram. 100 mL(-1). min(-1)), an endothelium-dependent or -independent vasodilator, respectively. The NO pathway was evaluated by repeating acetylcholine during L-arginine (200 microgram. 100 mL(-1). min(-1); 13 control subjects and 9 patients) or N(G)-monomethyl-L-arginine (L-NMMA; 100 microgram. 100 mL(-1). min(-1); 13 control subjects and 9 patients); production of cyclooxygenase-derived vasoconstrictors was assessed by repeating acetylcholine during indomethacin (50 microgram. 100 mL(-1). min(-1); 13 control subjects and 9 patients) or vitamin C (8 mg. 100 mL(-1). min(-1); 13 control subjects and 9 patients). Patients repeated the study within 1 month after ovariectomy and again after 3 months of estrogen replacement therapy (ERT; 17 beta-estradiol TTS, 50 microgram/d). Basally, vasodilation to acetylcholine was potentiated and inhibited by L-arginine and L-NMMA, respectively (P<0.05), but was unaffected by indomethacin or vitamin C. After ovariectomy, the modulating effect of L-arginine and L-NMMA disappeared, whereas indomethacin and vitamin C potentiated the response to acetylcholine (P<0.05). ERT restored L-arginine and L-NMMA effects on vasodilation to acetylcholine but prevented the potentiation caused by indomethacin or vitamin C. Response to sodium nitroprusside was unaffected by either ovariectomy or ERT. CONCLUSIONS: Endothelial dysfunction secondary to acute endogenous estrogen deprivation is caused by reduced NO availability. Cyclooxygenase-dependent production of oxidative stress could be responsible for this alteration. PMID- 10811593 TI - Vascular extracellular superoxide dismutase activity in patients with coronary artery disease: relation to endothelium-dependent vasodilation. AB - BACKGROUND: Increased inactivation of nitric oxide by oxygen free radicals contributes to endothelial dysfunction in patients with coronary artery disease (CAD). We therefore determined the activity of extracellular superoxide dismutase (EC-SOD), the major antioxidant enzyme system of the vessel wall, and its relation to flow-dependent, endothelium-mediated dilation (FDD) in patients with CAD. METHODS AND RESULTS: SOD isoenzyme activity was determined in coronary arteries from 10 patients with CAD and 10 control subjects. In addition, endothelium-bound EC-SOD activity (eEC-SOD), released by heparin bolus injection, and FDD of the radial artery were measured in 35 patients with CAD and 15 control subjects. FDD, determined by high-resolution ultrasound, was assessed at baseline, after intra-arterial infusion of vitamin C, N-monomethyl-L-arginine, and combination of both. EC-SOD activity in coronary arteries (control subjects: 126+/-14; CAD: 63+/-11 U/mg protein; P<0.01) and eEC-SOD activity in vivo (control subjects: 14.5+/-1.1; CAD: 3.8+/-1.1 U. mL(-1). min(-1); P<0.01) were reduced in patients with CAD. Activity of eEC-SOD was positively correlated with FDD (r=0.47; P<0. 01) and negatively with the effect of the antioxidant vitamin C on FDD (r=-0.59; P<0.01). In young individuals with hypercholesterolemia, however, eEC-SOD activity was increased (21. 0+/-1.2 U. mL(-1). min(-1); n=10; P<0.05). CONCLUSIONS: In patients with CAD, vascular EC-SOD activity is substantially reduced. The close relation between endothelium-bound EC-SOD activity and FDD suggests that reduced EC-SOD activity contributes to endothelial dysfunction in patients with CAD. In young hypercholesterolemic individuals, however, endothelium-bound EC-SOD activity is increased and may, in part, counteract impairment of endothelial function as the result of increased formation of oxygen free radicals. PMID- 10811594 TI - Impact of diabetes on cardiac structure and function: the strong heart study. AB - BACKGROUND: Whether diabetes mellitus (DM) adversely affects left ventricular (LV) structure and function independently of increases in body mass index (BMI) and blood pressure is controversial. METHODS AND RESULTS: Echocardiography was used in the Strong Heart Study, a study of cardiovascular disease in American Indians, to compare LV measurements between 1810 participants with DM and 944 with normal glucose tolerance. Participants with DM were older (mean age, 60 versus 59 years), had higher BMI (32.4 versus 28.9 kg/m(2)) and systolic blood pressure (133 versus 124 mm Hg), and were more likely to be female, to be on antihypertensive treatment, and to live in Arizona (all P<0.001). In analyses adjusted for covariates, women and men with DM had higher LV mass and wall thicknesses and lower LV fractional shortening, midwall shortening, and stress corrected midwall shortening (all P<0.002). Pulse pressure/stroke volume, a measure of arterial stiffness, was higher in participants with DM (P<0.001 independent of confounders). CONCLUSIONS: Non-insulin-dependent DM has independent adverse cardiac effects, including increased LV mass and wall thicknesses, reduced LV systolic chamber and myocardial function, and increased arterial stiffness. These findings identify adverse cardiovascular effects of DM, independent of associated increases in BMI and arterial pressure, that may contribute to cardiovascular events in diabetic individuals. PMID- 10811595 TI - Modulation of sympathetic coronary vasoconstriction by cardiac renin-angiotensin system in human coronary heart disease. AB - BACKGROUND: In humans, angiotensin II enhances the sympathetic coronary vasoconstriction elicited by the cold pressor test (CPT) and diving. Whether this enhancement depends on the circulating angiotensin II or on the locally produced angiotensin II is unknown, however. METHODS AND RESULTS: We addressed this issue in 14 patients with severe coronary artery disease by evaluating the effects of a 2-minute CPT (n=14) and a 30-second dive (n=8) on mean arterial pressure (MAP, arterial catheter), heart rate (ECG), coronary sinus blood flow (CBF, thermodilution technique), and coronary vascular resistance (MAP/CBF ratio). The 2 stimuli were applied at the end of left intracoronary infusion of either saline or benazeprilat diluted at the concentration of 25 microgram/mL. The rate of benazeprilat infusion had been preliminarily demonstrated to reduce angiotensin II concentration in the coronary sinus without affecting its arterial concentration. The changes in MAP and heart rate induced by CPT and diving were superimposable during saline and benazeprilat infusions. The decrease in CBF induced by CPT and diving during saline infusion was changed into an increase during benazeprilat infusion with a significant attenuation of the coronary vasoconstrictor response. CONCLUSIONS: In patients with coronary artery disease, an attenuation of sympathetic coronary vasoconstriction can be obtained by reducing cardiac angiotensin II formation without involving circulating angiotensin II. This suggests a role of the tissue renin-angiotensin system in modulating autonomic cardiac drive in humans. PMID- 10811596 TI - Racial differences in hemodynamic responses to environmental thermal stress among adolescents. AB - BACKGROUND: Previous studies by our laboratory and others have shown that blood pressure (BP) responses to many short-term laboratory stressors are greater in black than in white children. We sought to determine the cardiac and vascular contributions to these differences in BP reactivity and whether racial differences in vascular reactivity involve excessive vasoconstriction or deficient vasodilation. METHODS AND RESULTS: We evaluated BP, heart rate, and impedance cardiographic measures of preejection period (PEP) and total peripheral resistance (TPR) in healthy black (n=76) and white (n=60) adolescents (mean age, 14.8 years) during passive exposure to a vasoconstrictive cold chamber (8 degrees C to 10 degrees C) and a vasodilatory heat chamber (40 degrees C to 42 degrees C). Results indicated greater decreases in PEP and increases in TPR in blacks than whites during cold exposure (P<0.05) but no group differences during heat exposure. Covariance analyses indicated that the racial differences during cold exposure probably reflected greater beta-adrenergic cardiac reactivity and alpha adrenergic vasoconstrictive reactivity in blacks than whites. CONCLUSIONS: Blacks and whites exhibited comparable myocardial and vasodilatory responses to heat stress, but blacks exhibited heightened myocardial and vasoconstrictive reactivity to cold stress. These results suggest that the locus of racial differences in vascular reactivity involves vasoconstrictive rather than vasodilatory function. The pattern of racial differences during cold stress raises the possibility that both myocardial and vasoconstrictive mechanisms may contribute to the increased risk of hypertension in blacks. PMID- 10811597 TI - Prominent role of P-selectin in the development of advanced atherosclerosis in ApoE-deficient mice. AB - BACKGROUND: Adhesive interactions between leukocytes and endothelial cells are characteristic of the development of atherosclerotic lesions, but the receptors involved remain to be defined. P-selectin is an adhesion receptor expressed on activated endothelial cells or platelets and was shown to be involved in fatty streak formation in LDL receptor-deficient mice on an atherogenic diet. The main purpose of this study is to examine the role of P-selectin in the spontaneous development of advanced atherosclerosis in apoE-deficient mice. METHODDS AND RESULTS: We intercrossed P-selectin-deficient mice with mice lacking apoE and compared lesion development in apoE-deficient mice with P-selectin (apoE(-/-) P(+/+)) and without P-selectin (apoE(-/-) P(-/-)) that were fed normal mouse chow. At 4 months of age, apoE(-/-) P(-/-) mice had 3. 5-fold smaller aortic sinus lesions than apoE(-/-) P(+/+) mice. These were limited to fatty streaks in the apoE(-/-) P(-/-) mice, whereas 70% of apoE(-/-) P(+/+) lesions contained smooth muscle cells. Significantly more of the aortic sinus circumference was covered by lesions in the apoE(-/-) P(+/+) animals. The P-selectin genotype affected macrophage recruitment, because twice as many mononuclear cells were present in the P-selectin-positive lesions. At 15 months, the lesions progressed to the fibrous plaque stage in both genotypes and spread throughout the aorta, but this process was delayed in apoE(-/-) P(-/-) mice. In the aortic sinus, the lesions of the apoE(-/-) P(-/-) mice were 2.6-fold smaller and less calcified. CONCLUSIONS: P-selectin appears to be a key adhesion receptor mediating leukocyte recruitment into lesions and promoting advanced atherosclerosis in apoE-deficient mice. PMID- 10811598 TI - Effect of 40-kHz ultrasound on acute thrombotic ischemia in a rabbit femoral artery thrombosis model: enhancement of thrombolysis and improvement in capillary muscle perfusion. AB - BACKGROUND: We have shown previously that 40-kHz ultrasound (US) at low intensity accelerates fibrinolysis in vitro with little heating and good tissue penetration. These studies have now been extended to examine the effects of 40 kHz US on thrombolysis and tissue perfusion in a rabbit model. METHODS AND RESULTS: Treatment was administered with either US alone at 0.75 W/cm(2), streptokinase alone, or the combination of US and streptokinase. US or streptokinase resulted in minimal thrombolysis, but reperfusion was nearly complete with the combination after 120 minutes. US also reversed the ischemia in nonperfused muscle in the absence of arterial flow. Tissue perfusion decreased after thrombosis from 13. 7+/-0.2 to 6.6+/-0.8 U and then declined further to 4.5+/-0.4 U after 240 minutes. US improved perfusion to 10.6+/-0.5 and 12.1+/-0. 5 U after 30 and 60 minutes, respectively. This effect was reversible and declined to pretreatment values after US was discontinued. Similarly, tissue pH declined from normal to 7.05+/-0. 02 after thrombosis, but US improved pH to 7.34+/-0.03 after 60 minutes. US-induced improvement in tissue perfusion and pH also occurred after femoral artery ligation, indicating that thrombolysis did not cause these effects. CONCLUSIONS: 40-kHz US at low intensity markedly accelerates fibrinolysis and also improves tissue perfusion and reverses acidosis, effects that would be beneficial in treatment of acute thrombosis. PMID- 10811599 TI - Dopamine as a novel antioxidative agent for rat vascular smooth muscle cells through dopamine D(1)-like receptors. AB - BACKGROUND: To elucidate the roles of vascular D(1)-like receptors in atherosclerosis, the effects of the specific D(1)-like agonists on platelet derived growth factor (PDGF)-BB-mediated oxidative stress in vascular smooth muscle cells (VSMCs) were studied. METHODS AND RESULTS: Immunohistochemical studies demonstrated the coexistence of D(1A) and D(1B) dopamine receptors in VSMCs. Western blotting revealed a band of approximately 70 kDa for D(1A) and D(1B) dopamine receptors. VSMCs stimulated by PDGF-BB exhibited increased oxidative stress directly measured by flow cytometry. These effects were prevented by dopamine, SKF 38393, or YM 435, and this prevention was reversed by Sch 23390. These effects were blocked by a specific protein kinase A (PKA) inhibitor, N-(2-[p-bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide (H 89). The PDGF-BB-mediated increase in oxidative stress of VSMCs was significantly suppressed by the indirect phospholipase D (PLD) inhibitor suramin or the specific protein kinase C (PKC) inhibitor calphostin C. Both antisense but neither sense nor scrambled oligonucleotides to D(1A) and D(1B) receptors inhibited dopamine-induced suppression of increase in oxidative stress of VSMCs induced by PDGF-BB. CONCLUSIONS: These findings suggest that vascular D(1)-like receptors (D(1A) and D(1B) receptors) inhibit any increase in oxidative stress of VSMCs, possibly through activation of PKA and suppression of PLD and PKC. PMID- 10811600 TI - Hypotension and resistance to lipopolysaccharide-induced shock in transgenic mice overexpressing adrenomedullin in their vasculature. AB - BACKGROUND: Adrenomedullin (AM) is a vasodilating peptide involved in the regulation of circulatory homeostasis and in the pathophysiology of certain cardiovascular diseases. To determine the extent to which chronic AM overproduction affects circulatory physiology under normal and pathological conditions, we used a preproendothelin-1 promoter to establish transgenic mouse lines overexpressing AM in their vasculature. METHODS AND RESULTS: Transgenic mice overexpressing AM mainly in vascular endothelial and smooth muscle cells exhibited significantly lower blood pressure (BP) and higher plasma cGMP levels than their wild-type littermates. Blockade of NO synthase with N(G)-monomethyl-L arginine elevated BP to a greater degree in AM transgenic mice, offsetting the BP difference between the 2 groups. Despite their lower basal BP, administration of bacterial lipopolysaccharide elicited smaller declines in BP and less severe organ damage in AM transgenic mice than in wild-type mice. Furthermore, the 24 hour survival rate after induction of lipopolysaccharide shock was significantly higher in the transgenic mice. CONCLUSIONS: A chronic increase in vascular AM production reduces BP at least in part via an NO-dependent pathway. In addition, smaller responses to LPS in transgenic mice suggest that AM is protective against the circulatory collapse, organ damage, and mortality characteristic of endotoxic shock. PMID- 10811601 TI - Angiopoietin-1 inhibits irradiation- and mannitol-induced apoptosis in endothelial cells. AB - BACKGROUND AND PURPOSE: Angiopoietin-1 (Ang1) is a vasculogenic factor that signals through the endothelial cell-specific Tie2 receptor tyrosine kinase. We recently reported that Ang1 prevented apoptosis induced by serum deprivation in endothelial cells. In this study, we examined whether Ang1 prevents apoptosis in endothelial cells treated with irradiation or clinical concentrations of mannitol. METHODS AND RESULTS: ++Ang1 prevented irradiation- and mannitol-induced apoptosis in human umbilical vein endothelial cells in a dose-dependent manner. Pretreatment with soluble Tie2 receptor, but not Tie1 receptor, blocked the antiapoptotic effect of Ang1. Two phosphatidylinositol 3'-kinase (PI3-kinase) specific inhibitors, wortmannin and LY294002, blocked the Ang1-induced antiapoptotic effect. The antiapoptotic potency of Ang1 was similar to or greater than that of vascular endothelial growth factor, basic fibroblast growth factor, and endothelin-1. Ang1 also prevented apoptosis in cultured endothelial cells from porcine pulmonary and coronary arteries and in endothelial cells of explanted rat aorta. CONCLUSIONS: Ang1 promotes the survival of endothelial cells in irradiation- and mannitol-induced apoptosis through Tie2 receptor binding and PI3-kinase activation. Pretreatment with Ang1 could be beneficial in maintaining normal endothelial cell integrity during intracoronary irradiation or systemic mannitol therapy. PMID- 10811602 TI - Closing the gap in quality health care for Americans. PMID- 10811603 TI - Images in cardiovascular medicine. Three-dimensional observation of the intracellular membrane structure in human myocardium: high-resolution scanning electron microscopy by the osmium-DMSO-osmium method. PMID- 10811604 TI - Acute cardiac tamponade caused by massive hemorrhage from pericardial cyst. PMID- 10811605 TI - The Crd1 gene encodes a putative di-iron enzyme required for photosystem I accumulation in copper deficiency and hypoxia in Chlamydomonas reinhardtii. AB - Chlamydomonas reinhardtii adapts to copper deficiency by degrading apoplastocyanin and inducing Cyc6 and Cpx1 encoding cytochrome c(6) and coproporphyrinogen oxidase, respectively. To identify other components in this pathway, colonies resulting from insertional mutagenesis were screened for copper conditional phenotypes. Twelve crd (copper response defect) strains were identified. In copper-deficient conditions, the crd strains fail to accumulate photosystem I and light-harvesting complex I, and they contain reduced amounts of light-harvesting complex II. Cyc6, Cpx1 expression and plastocyanin accumulation remain copper responsive. The crd phenotype is rescued by a similar amount of copper as is required for repression of Cyc6 and Cpx1 and for maintenance of plastocyanin at its usual stoichiometry, suggesting that the affected gene is a target of the same signal transduction pathway. The crd strains represent alleles at a single locus, CRD1, which encodes a 47 kDa, hydrophilic protein with a consensus carboxylate-bridged di-iron binding site. Crd1 homologs are present in the genomes of photosynthetic organisms. In Chlamydomonas, Crd1 expression is activated in copper- or oxygen-deficient cells, and Crd1 function is required for adaptation to these conditions. PMID- 10811606 TI - Structure of the bacteriorhodopsin mutant F219L N intermediate revealed by electron crystallography. AB - Bacteriorhodopsin is a light-driven proton pump in halobacteria that forms crystalline patches in the cell membrane. Isomerization of the bound retinal initiates a photocycle resulting in the extrusion of a proton. An electron crystallographic analysis of the N intermediate from the mutant F219L gives a three-dimensional view of the large conformational change that occurs on the cytoplasmic side after deprotonation of the retinal Schiff base. Helix F, together with helix E, tilts away from the center of the molecule, causing a shift of approximately 3 A at the EF loop. The top of helix G moves slightly toward the ground state location of helix F. These movements open a water accessible channel in the protein, enabling the transfer of a proton from an aspartate residue to the Schiff base. The movement of helix F toward neighbors in the crystal lattice is so large that it would not allow all molecules to change conformation simultaneously, limiting the occupancy of this state in the membrane to 33%. This explains photocooperative phenomena in the purple membrane. PMID- 10811607 TI - Ubc8p functions in catabolite degradation of fructose-1, 6-bisphosphatase in yeast. AB - The key gluconeogenic enzyme fructose-1,6-bisphosphatase (FBPase) is synthesized when cells of the yeast Saccharomyces cerevisiae are grown on a non-fermentable carbon source. After shifting the cells to glucose-containing medium, in a process called catabolite degradation, FBPase is selectively and rapidly broken down. We have isolated gid mutants, which are defective in this glucose-induced degradation process. When complementing the defect in catabolite degradation of FBPase in gid3-1 mutant cells with a yeast genomic library, we identified the GID3 gene and found it to be identical to UBC8 encoding the ubiquitin-conjugating enzyme Ubc8p. The in vivo function of Ubc8p (Gid3p) has remained a mystery so far. Here we demonstrate the involvement of Ubc8p in the glucose-induced ubiquitylation of FBPase as a prerequisite for catabolite degradation of the enzyme via the proteasome. Like FBPase, Ubc8p is found in the cytoplasmic fraction of the cell. We demonstrate cytoplasmic degradation of FBPase. PMID- 10811608 TI - Cyclin E-mediated elimination of p27 requires its interaction with the nuclear pore-associated protein mNPAP60. AB - The Cdk2 inhibitor, p27(Kip1), is degraded in a phosphorylation- and ubiquitylation-dependent manner at the G(1)-S transition of the cell cycle. Degradation of p27(Kip1) requires import into the nucleus for phosphorylation by Cdk2. Phosphorylated p27(Kip1) is thought to be subsequently re-exported and degraded in the cytosol. Using two-hybrid screens, we now show that p27(Kip1) interacts with a nuclear pore-associated protein, mNPAP60, map the interaction to the 3(10) helix of p27 and identify a point mutant in p27(Kip1) that is deficient for interaction (R90G). In vivo and in vitro, the loss-of-interaction mutant is poorly transported into the nucleus, while ubiquitylation of p27R90G occurs normally. In vivo, co-expression of cyclin E and Cdk2 rescues the import defect. However, mutant p27(Kip1) accumulates in a phosphorylated form in the nucleus and is not efficiently degraded, arguing that at least one step in the degradation of phosphorylated p27(Kip1) requires an interaction with the nuclear pore. Our results identify a novel component involved in p27(Kip1) degradation and suggest that degradation of p27(Kip1) is tightly linked to its intracellular transport. PMID- 10811609 TI - A complex of mammalian ufd1 and npl4 links the AAA-ATPase, p97, to ubiquitin and nuclear transport pathways. AB - The AAA-ATPase, p97/Cdc48p, has been implicated in many different pathways ranging from membrane fusion to ubiquitin-dependent protein degradation. Binding of the p47 complex directs p97 to act in the post-mitotic fusion of Golgi membranes. We now describe another binding complex comprising mammalian Ufd1 and Npl4. Yeast Ufd1p is required for ubiquitin-dependent protein degradation whereas yeast Npl4p has been implicated in nuclear transport. In rat liver cytosol, Ufd1 and Npl4 form a binary complex, which exists either alone or bound to p97. Ufd1/Npl4 competes with p47 for binding to p97 and so inhibits Golgi membrane fusion. This suggests that it is involved in another cellular function catalysed by p97, the most likely being ubiquitin-dependent events during mitosis. The fact that the binding of p47 and Ufd1/Npl4 is mutually exclusive suggests that these protein complexes act as adapters, directing a basic p97 activity into different cellular pathways. PMID- 10811611 TI - Hsp47: a molecular chaperone that interacts with and stabilizes correctly-folded procollagen. AB - Hsp47 is a heat-shock protein that interacts transiently with procollagen during its folding, assembly and transport from the endoplasmic reticulum (ER) of mammalian cells. It has been suggested to carry out a diverse range of functions, such as acting as a molecular chaperone facilitating the folding and assembly of procollagen molecules, retaining unfolded molecules within the ER, and assisting the transport of correctly folded molecules from the ER to the Golgi apparatus. Here we define the substrate recognition of Hsp47, demonstrating that it interacts preferentially with triple-helical procollagen molecules. The association of Hsp47 with procollagen coincides with the formation of a collagen triple helix. This demonstrates that Hsp47's role in procollagen folding and assembly is distinct from that of prolyl 4-hydroxylase. These results indicate that Hsp47 acts as a novel molecular chaperone, potentially stabilizing the correctly folded collagen helix from heat denaturation before its transport from the ER. PMID- 10811610 TI - mu1A-adaptin-deficient mice: lethality, loss of AP-1 binding and rerouting of mannose 6-phosphate receptors. AB - The heterotetrameric AP-1 complex is involved in the formation of clathrin-coated vesicles at the trans-Golgi network (TGN) and interacts with sorting signals in the cytoplasmic tails of cargo molecules. Targeted disruption of the mouse mu1A adaptin gene causes embryonic lethality at day 13.5. In cells deficient in micro1A-adaptin the remaining AP-1 adaptins do not bind to the TGN. Polarized epithelial cells are the only cells of micro1A-adaptin-deficient embryos that show gamma-adaptin binding to membranes, indicating the formation of an epithelial specific AP-1B complex and demonstrating the absence of additional mu1A homologs. Mannose 6-phosphate receptors are cargo molecules that exit the TGN via AP-1-clathrin-coated vesicles. The steady-state distribution of the mannose 6-phosphate receptors MPR46 and MPR300 in mu1A-deficient cells is shifted to endosomes at the expense of the TGN. MPR46 fails to recycle back from the endosome to the TGN, indicating that AP-1 is required for retrograde endosome to TGN transport of the receptor. PMID- 10811612 TI - The propeptide of macrophage inhibitory cytokine (MIC-1), a TGF-beta superfamily member, acts as a quality control determinant for correctly folded MIC-1. AB - Macrophage inhibitory cytokine (MIC-1), a divergent member of the transforming growth factor-beta (TGF-beta) superfamily and activation associated cytokine, is secreted as a 28 kDa dimer. To understand its secretion, we examined its processing in MIC-1-transfected Chinese hamster ovary cells. Mature MIC-1 dimer arises post-endoplasmic reticulum (ER) by proteolytic cleavage of dimeric pro-MIC 1 precursor at a furin-like site. Unlike previously characterized TGF-beta superfamily members, MIC-1 dimers are also secreted in constructs lacking the propeptide. A clue to the function of the propeptide came from the observation that a range of proteasome inhibitors, including lactacystin and MG132, cause major increases in levels of undimerized pro-MIC-1 precursor. There was no effect of proteasome inhibitors on cells expressing mature MIC-1 without the propeptide, suggesting that the propeptide can signal misfolding of MIC-1, leading to proteasomal degradation. Deletion mutagenesis showed the N-terminal 28 amino acids of the propeptide are necessary for proteasomal degradation. This is the first demonstration, to our knowledge, of a quality control function in a propeptide domain of a secretory protein and represents an additional mechanism to ensure correct folding of proteins leaving the ER. PMID- 10811613 TI - Pilus formation and protein secretion by the same machinery in Escherichia coli. AB - The secreton (type II secretion) and type IV pilus biogenesis branches of the general secretory pathway in Gram-negative bacteria share many features that suggest a common evolutionary origin. Five components of the secreton, the pseudopilins, are similar to subunits of type IV pili. Here, we report that when the 15 genes encoding the pullulanase secreton of Klebsiella oxytoca were expressed on a high copy number plasmid in Escherichia coli, one pseudopilin, PulG, was assembled into pilus-like bundles. Assembly of the 'secreton pilus' required most but not all of the secreton components that are essential for pullulanase secretion, including some with no known homologues in type IV piliation machineries. Two other pseudopilins, pullulanase and two outer membrane associated secreton components were not associated with pili. Thus, PulG is probably the major component of the pilus. Expression of a type IV pilin gene, the E.coli K-12 gene ppdD, led to secreton-dependent incorporation of PpdD pilin into pili without diminishing pullulanase secretion. This is the first demonstration that pseudopilins can be assembled into pilus-like structures. PMID- 10811614 TI - Domain structure of secretin PulD revealed by limited proteolysis and electron microscopy. AB - Secretins, a superfamily of multimeric outer membrane proteins, mediate the transport of large macromolecules across the outer membrane of Gram-negative bacteria. Limited proteolysis of secretin PulD from the Klebsiella oxytoca pullulanase secretion pathway showed that it consists of an N-terminal domain and a protease-resistant C-terminal domain that remains multimeric after proteolysis. The stable C-terminal domain starts just before the region in PulD that is highly conserved in the secretin superfamily and apparently lacks the region at the C terminal end to which the secretin-specific pilot protein PulS binds. Electron microscopy showed that the stable fragment produced by proteolysis is composed of two stacked rings that encircle a central channel and that it lacks the peripheral radial spokes that are seen in the native complex. Moreover, the electron microscopic images suggest that the N-terminal domain folds back into the large cavity of the channel that is formed by the C-terminal domain of the native complex, thereby occluding the channel, consistent with previous electrophysiological studies showing that the channel is normally closed. PMID- 10811615 TI - Protein phosphatase 1alpha is a Ras-activated Bad phosphatase that regulates interleukin-2 deprivation-induced apoptosis. AB - Growth factor deprivation is a physiological mechanism to regulate cell death. We utilize an interleukin-2 (IL-2)-dependent murine T-cell line to identify proteins that interact with Bad upon IL-2 stimulation or deprivation. Using the yeast two hybrid system, glutathione S-transferase (GST) fusion proteins and co immunoprecipitation techniques, we found that Bad interacts with protein phosphatase 1alpha (PP1alpha). Serine phosphorylation of Bad is induced by IL-2 and its dephosphorylation correlates with appearance of apoptosis. IL-2 deprivation induces Bad dephosphorylation, suggesting the involvement of a serine phosphatase. A serine/threonine phosphatase activity, sensitive to the phosphatase inhibitor okadaic acid, was detected in Bad immunoprecipitates from IL-2-stimulated cells, increasing after IL-2 deprivation. This enzymatic activity also dephosphorylates in vivo (32)P-labeled Bad. Treatment of cells with okadaic acid blocks Bad dephosphorylation and prevents cell death. Finally, Ras activation controls the catalytic activity of PP1alpha. These results strongly suggest that Bad is an in vitro and in vivo substrate for PP1alpha phosphatase and that IL-2 deprivation-induced apoptosis may operate by regulating Bad phosphorylation through PP1alpha phosphatase, whose enzymatic activity is regulated by Ras. PMID- 10811616 TI - A temperature-sensitive adenylyl cyclase mutant of Dictyostelium. AB - Dictyostelium development starts with the chemotactic aggregation of up to 10(6) amoebae in response to propagating cAMP waves. cAMP is produced by the aggregation stage adenylyl cyclase (ACA) and cells lacking ACA (aca null) cannot aggregate. Temperature-sensitive mutants of ACA were selected from a population of aca null cells transformed with a library of ACA genes, a major segment of which had been amplified by error-prone PCR. One mutant (tsaca2) that can complement the aggregation null phenotype of aca null cells at 22 degrees C but not at 28 degrees C was characterized in detail. The basal catalytic activity of the enzyme in this mutant was rapidly and reversibly inactivated at 28 degrees C. Using this mutant strain we show that cell movement in aggregates and mounds is organized by propagating waves of cAMP. Synergy experiments between wild-type and tsaca2 cells, shifted to the restrictive temperature at various stages of development, showed that ACA plays an important role in the control of cell sorting and tip formation. PMID- 10811617 TI - Thr38 and Ser198 are Pto autophosphorylation sites required for the AvrPto-Pto mediated hypersensitive response. AB - The tomato Pto kinase confers resistance to Pseudomonas syringae pv. tomato expressing the AvrPto protein. To elucidate the role of Pto autophosphorylation in disease resistance, eight sites autophosphorylated by Pto in vitro were identified by a combination of HPLC purification of tryptic phosphopeptides, MALDI-TOF/MS analysis and Edman degradation. Mutational analysis of the autophosphorylation sites revealed that Pto residues Thr38 and Ser198 are required for AvrPto-Pto- mediated elicitation of a hypersensitive response in the plant. Thr38, which is the main Pto autophosporylation site and is located outside the kinase catalytic domain, was also required for Pto kinase activity and its physical interaction with AvrPto, the Pti1 kinase and the transcription factor Pti4. Ser198, located in the Pto activation domain, was dispensable for kinase activity and for interaction with AvrPto. However, a mutation at this site resulted in altered Pto interactions with the Pti1 kinase and the Pto interactors of unknown function Pti3 and Pti10. These results suggest that autophosphorylation events at Pto Thr38 and Ser198 are required for signal transduction by Pto and participate in distinct molecular mechanisms. PMID- 10811618 TI - Structural basis of the Axin-adenomatous polyposis coli interaction. AB - Axin and the adenomatous polyposis coli (APC) tumor suppressor protein are components of the Wnt/Wingless growth factor signaling pathway. In the absence of Wnt signal, Axin and APC regulate cytoplasmic levels of the proto-oncogene beta catenin through the formation of a large complex containing these three proteins, glycogen synthase kinase 3beta (GSK3beta) and several other proteins. Both Axin and APC are known to be critical for beta-catenin regulation, and truncations in APC that eliminate the Axin-binding site result in human cancers. A protease resistant domain of Axin that contains the APC-binding site is a member of the regulators of G-protein signaling (RGS) superfamily. The crystal structures of this domain alone and in complex with an Axin-binding sequence from APC reveal that the Axin-APC interaction occurs at a conserved groove on a face of the protein that is distinct from the G-protein interface of classical RGS proteins. The molecular interactions observed in the Axin-APC complex provide a rationale for the evolutionary conservation seen in both proteins. PMID- 10811619 TI - The sno gene, which encodes a component of the histone deacetylase complex, acts as a tumor suppressor in mice. AB - The Ski and Sno oncoproteins are components of a macromolecular complex containing the co-repressor N-CoR/SMRT, mSin3 and histone deacetylase. This complex has been implicated in the transcriptional repression exerted by a number of repressors including nuclear hormone receptors and Mad. Further more, Ski and Sno negatively regulate transforming growth factor-beta (TGF-beta) signaling by recruiting this complex to Smads. Here we show that loss of one copy of sno increases susceptibility to tumorigenesis in mice. Mice lacking sno died at an early stage of embryogenesis, and sno was required for blastocyst formation. Heterozygous (sno(+/-)) mice developed spontaneous lymphomas at a low frequency and showed an increased level of tumor formation relative to wild-type mice when challenged with a chemical carcinogen. sno(+/-) embryonic fibroblasts had an increased proliferative capacity and the introduction of activated Ki-ras into these cells resulted in neoplastic transformation. The B cells, T cells and embryonic fibroblasts of sno(+/-) mice had a decreased sensitivity to apoptosis or cell cycle arrest. These findings demonstrate that sno acts as a tumor suppressor at least in some types of cells. PMID- 10811620 TI - Transcriptional repression by Pax5 (BSAP) through interaction with corepressors of the Groucho family. AB - Pax5 (BSAP) functions as both a transcriptional activator and repressor during midbrain patterning, B-cell development and lymphomagenesis. Here we demonstrate that Pax5 exerts its repression function by recruiting members of the Groucho corepressor family. In a yeast two-hybrid screen, the groucho-related gene product Grg4 was identified as a Pax5 partner protein. Both proteins interact cooperatively via two separate domains: the N-terminal Q and central SP regions of Grg4, and the octapeptide motif and C-terminal transactivation domain of Pax5. The phosphorylation state of Grg4 is altered in vivo upon Pax5 binding. Moreover, Grg4 efficiently represses the transcriptional activity of Pax5 in an octapeptide dependent manner. Similar protein interactions resulting in transcriptional repression were also observed between distantly related members of both the Pax2/5/8 and Groucho protein families. In agreement with this evolutionary conservation, the octapeptide motif of Pax proteins functions as a Groucho dependent repression domain in Drosophila embryos. These data indicate that Pax proteins can be converted from transcriptional activators to repressors through interaction with corepressors of the Groucho protein family. PMID- 10811621 TI - A bacterial sensor of plant cell contact controls the transcriptional induction of Ralstonia solanacearum pathogenicity genes. AB - The hrp genes of the plant pathogen Ralstonia solanacearum are key pathogenicity determinants; they encode a type III protein secretion machinery involved in the secretion of mediators of the bacterium-plant interaction. These hrp genes are under the genetic control of the hrpB regulatory gene, expression of which is induced when bacteria are co-cultivated with plant cell suspensions. In this study, we used hrp-gfp transcriptional fusions to demonstrate that the expression of the hrpB and type III secretion genes is specifically induced in response to the bacterium-plant cell contact. This contact-dependent induction of hrpB gene expression requires the outer membrane protein PrhA, but not a functional type III secretion apparatus. Genetic evidence indicates that PrhA constitutes the first example of a bacterial receptor for a non-diffusible signal present in the plant cell wall and which triggers the transcriptional activation of bacterial virulence genes. PMID- 10811622 TI - Structural and functional conservation at the boundaries of the chicken beta globin domain. AB - We show that the 3' boundary of the chicken beta-globin locus bears striking structural similarities to the 5' boundary. In erythroid cells a clear transition in DNase I sensitivity of chromatin at the 3' end of the locus is observed, the location of this transition is marked by a constitutive DNase I hypersensitive site (HS), and DNA spanning this site has the enhancer-blocking capacity of an insulator. This HS contains a binding site for the transcription factor CTCF. As in the case of the 5' insulator, the CTCF site is both necessary and sufficient for the enhancer-blocking activity of the 3' boundary. The position of this insulator is consistent with our proposal that it may function to maintain the distinct regulatory programs of the globin genes and their closely appended 3' neighbor, an odorant receptor gene. We conclude that both boundaries of the chicken beta-globin domain are capable of playing functionally similar roles and that the same protein is a necessary component of the molecular mechanism through which these boundaries are defined. PMID- 10811623 TI - The chromo domain protein chd1p from budding yeast is an ATP-dependent chromatin modifying factor. AB - CHD proteins are members of the chromo domain family, a class of proteins involved in transcription, DNA degradation and chromatin structure. In higher eukaryotes, there are two distinct subfamilies of CHD proteins: CHD1 and CHD3/4. Analyses carried out in vitro indicate that the CHD3/4 proteins may regulate transcription via alteration of chromatin structure. However, little is known about the role of CHD proteins in vivo, particularly the CHD1 subfamily. To understand better the cellular function of CHD proteins, we initiated a study on the Chd1p protein from budding yeast. Using genomic DNA arrays, we identified genes whose expression is affected by the absence of Chd1p. A synthetic-lethal screen uncovered genetic interactions between SWI/SNF genes and CHD1. Biochemical experiments using Chd1p purified from yeast showed that it reconfigures the structure of nucleosome core particles in a manner distinct from the SWI-SNF complex. Taken together, these results suggest that Chd1p functions as a nucleosome remodeling factor, and that Chd1p may share overlapping roles with the SWI-SNF complex to regulate transcription. PMID- 10811624 TI - Telomere maintenance and length regulation in Trypanosoma brucei. AB - Transcription of telomere proximal variant surface glycoprotein genes is mono allelic in bloodstream-form Trypanosoma brucei. The terminal DNA sequence at these telomeres consists of tandem T(2)AG(3) repeats, which increase in length by approximately 8 bp per cell division balanced by occasional loss of large numbers of repeats. Here we have used targeted chromosome fragmentation to investigate the sequence requirements for telomere formation in T. brucei. Telomere formation is most efficient on tandem T(2)AG(3) repeats, but can also occur on specific templates found within 'random' sequence substrates and on G-rich motifs proximal to a double-strand break. Newly formed telomeres are extended faster than other native telomeres, but as the telomere becomes longer the rate of extension declines. Telomere length regulation in T.brucei is discussed in the context of recent results from other cell types. PMID- 10811625 TI - HuR regulates cyclin A and cyclin B1 mRNA stability during cell proliferation. AB - Colorectal carcinoma RKO cells expressing reduced levels of the RNA-binding protein HuR (ASHuR) displayed markedly reduced growth. In synchronous RKO populations, HuR was almost exclusively nuclear during early G(1), increasing in the cytoplasm during late G(1), S and G(2). The expression and half-life of mRNAs encoding cyclins A and B1 similarly increased during S and G(2), then declined, indicating that mRNA stabilization contributed to their cell cycle-regulated expression. In gel-shift assays using radiolabeled cyclin RNA transcripts and RKO protein extracts, only those transcripts corresponding to the 3'-untranslated regions of cyclins A and B1 formed RNA-protein complexes in a cell cycle dependent fashion. HuR directly bound mRNAs encoding cyclins A and B1, as anti HuR antibodies supershifted such RNA-protein complexes. Importantly, the expression and half-life of mRNAs encoding cyclins A and B1 were reduced in ASHuR RKO cells. Our results indicate that HuR may play a critical role in cell proliferation, at least in part by mediating cell cycle-dependent stabilization of mRNAs encoding cyclins A and B1. PMID- 10811626 TI - The 2 A crystal structure of leucyl-tRNA synthetase and its complex with a leucyl adenylate analogue. AB - Leucyl-, isoleucyl- and valyl-tRNA synthetases are closely related large monomeric class I synthetases. Each contains a homologous insertion domain of approximately 200 residues, which is thought to permit them to hydrolyse ('edit') cognate tRNA that has been mischarged with a chemically similar but non-cognate amino acid. We describe the first crystal structure of a leucyl-tRNA synthetase, from the hyperthermophile Thermus thermophilus, at 2.0 A resolution. The overall architecture is similar to that of isoleucyl-tRNA synthetase, except that the putative editing domain is inserted at a different position in the primary structure. This feature is unique to prokaryote-like leucyl-tRNA synthetases, as is the presence of a novel additional flexibly inserted domain. Comparison of native enzyme and complexes with leucine and a leucyl- adenylate analogue shows that binding of the adenosine moiety of leucyl-adenylate causes significant conformational changes in the active site required for amino acid activation and tight binding of the adenylate. These changes are propagated to more distant regions of the enzyme, leading to a significantly more ordered structure ready for the subsequent aminoacylation and/or editing steps. PMID- 10811627 TI - Crystal structure of the ribosome recycling factor from Escherichia coli. AB - We have determined the crystal structure of the Escherichia coli ribosome recycling factor (RRF), which catalyzes the disassembly of the termination complex in protein synthesis. The L-shaped molecule consists of two domains: a triple-stranded antiparallel coiled-coil and an alpha/beta domain. The coil domain has a cylindrical shape and negatively charged surface, which are reminiscent of the anticodon arm of tRNA and domain IV of elongation factor EF-G. We suggest that RRF binds to the ribosomal A-site through its coil domain, which is a tRNA mimic. The relative position of the two domains is changed about an axis along the hydrophobic cleft in the hinge where the alkyl chain of a detergent molecule is bound. The tRNA mimicry and the domain movement observed in RRF provide a structural basis for understanding the role of RRF in protein synthesis. PMID- 10811628 TI - A domain in the N-terminal extension of class IIb eukaryotic aminoacyl-tRNA synthetases is important for tRNA binding. AB - Cytoplasmic aspartyl-tRNA synthetase (AspRS) from Saccharomyces cerevisiae is a homodimer of 64 kDa subunits. Previous studies have emphasized the high sensitivity of the N-terminal region to proteolytic cleavage, leading to truncated species that have lost the first 20-70 residues but that retain enzymatic activity and dimeric structure. In this work, we demonstrate that the N terminal extension in yeast AspRS participates in tRNA binding and we generalize this finding to eukaryotic class IIb aminoacyl-tRNA synthetases. By gel retardation studies and footprinting experiments on yeast tRNA(Asp), we show that the extension, connected to the anticodon-binding module of the synthetase, contacts tRNA on the minor groove side of its anticodon stem. Sequence comparison of eukaryotic class IIb synthetases identifies a lysine-rich 11 residue sequence ((29)LSKKALKKLQK(39) in yeast AspRS with the consensus xSKxxLKKxxK in class IIb synthetases) that is important for this binding. Direct proof of the role of this sequence comes from a mutagenesis analysis and from binding studies using the isolated peptide. PMID- 10811629 TI - Recombination-induced CAG trinucleotide repeat expansions in yeast involve the MRE11-RAD50-XRS2 complex. AB - Recombination induced by double-strand breaks (DSBs) in yeast leads to a higher proportion of expansions to contractions than does replication-associated tract length changes. Expansions are apparently dependent on the property of the repeat array to form hairpins, since DSB repair of a CAA(87) repeat induces only contractions of the repeat sequence. DSB-repair efficiency is reduced by 40% when DNA synthesis must traverse a CAG(98) array, as compared with a CAA(87) array. These data indicate that repair- associated DNA synthesis is inhibited by secondary structures formed by CAG(98) and that these structures promote repeat expansions during DSB repair. Overexpression of Mre11p or Rad50p suppresses the inhibition of DSB repair by CAG(98) and significantly increases the average size of expansions found at the recipient locus. Both effects are dependent on the integrity of the Mre11p-Rad50p-Xrs2p complex. The Mre11 complex thus appears to be directly involved in removing CAG or CTG hairpins that arise frequently during DNA synthesis accompanying gene conversion of these trinucleotide repeats. PMID- 10811630 TI - Neuromedin U is a potent agonist at the orphan G protein-coupled receptor FM3. AB - Neuromedins are a family of peptides best known for their contractile activity on smooth muscle preparations. The biological mechanism of action of neuromedin U remains unknown, despite the fact that the peptide was first isolated in 1985. Here we show that neuromedin U potently activates the orphan G protein-coupled receptor FM3, with subnanomolar potency, when FM3 is transiently expressed in human HEK-293 cells. Neuromedins B, C, K, and N are all inactive at this receptor. Quantitative reverse transcriptase-polymerase chain reaction analysis of neuromedin U expression in a range of human tissues showed that the peptide is highly expressed in the intestine, pituitary, and bone marrow, with lower levels of expression seen in stomach, adipose tissue, lymphocytes, spleen, and the cortex. Similar analysis of FM3 expression showed that the receptor is widely expressed in human tissue with highest levels seen in adipose tissue, intestine, spleen, and lymphocytes, suggesting that neuromedin U may have a wide range of presently undetermined physiological effects. The discovery that neuromedin U is an endogenous agonist for FM3 will significantly aid the study of the full physiological role of this peptide. PMID- 10811631 TI - Osteoprotegerin is an alpha vbeta 3-induced, NF-kappa B-dependent survival factor for endothelial cells. AB - Osteopontin protects endothelial cells from apoptosis induced by growth factor withdrawal. This interaction is mediated by the alpha(v)beta(3) integrin and is NF-kappaB-dependent (Scatena, M., Almeida, M., Chaisson, M. L., Fausto, N., Nicosia, R. F., and Giachelli, C. M. (1998) J. Cell Biol. 141, 1083-1093). In the present study we used differential cloning to identify osteopontin-induced, NF kappaB-dependent genes in endothelial cells. One of the genes identified in this screen was osteoprotegerin, a member of the tumor necrosis factor receptor superfamily. By Northern and Western blot analysis, osteoprotegerin mRNA and protein levels were very low in endothelial cells plated on the non-integrin cell attachment factor, poly-d-lysine. In contrast, osteoprotegerin mRNA and protein levels were induced 5-7-fold following alpha(v)beta(3) ligation by osteopontin. Osteoprotegerin induction by osteopontin was time-dependent and observed as early as 3 h following treatment. NF-kappaB inactivation achieved by over expression of an IkappaB super repressor in endothelial cells completely inhibited osteoprotegerin induction by osteopontin. Finally, purified osteoprotegerin protected endothelial cells with inactive NF-kappaB from apoptosis induced by growth factor deprivation. These data suggest that alpha(v)beta(3)-mediated endothelial survival depends on osteoprotegerin induction by NF-kappaB and indicate a new function for osteoprotegerin in endothelial cells. PMID- 10811632 TI - Insulin-like growth factor I-mediated degradation of insulin receptor substrate-1 is inhibited by epidermal growth factor in prostate epithelial cells. AB - We have sought to determine whether insulin-like growth factor I (IGF-I) regulates the levels of insulin receptor substrate-1 (IRS-1) in prostate epithelial cells. Exposure of prostate epithelial cells to IGF-I in the absence of other growth factors leads to a reduction in IRS-1 levels. Ubiquitin content of IRS-1 is increased in the presence of IGF-I, and inhibitors of the proteasome prevented the reduction of IRS-1 levels seen following IGF-I exposure. These results imply that IRS-1 is targeted to the proteasome upon exposure to IGF-I. The addition of epidermal growth factor (EGF) maintained IRS-1 levels even in the presence of IGF-I and inhibits IGF-I-dependent ubiquitination of IRS-1. Thus, these two growth factors, IGF-I and EGF, had antagonistic effects on IRS-1 protein levels in prostate epithelial cells. This regulation of IRS-1 reveals a novel level of cross-talk between the IGF-I and EGF signal pathways, which may have implications in tumors that harbor activating mutations in the EGF receptor. PMID- 10811633 TI - Telomerase activity reconstituted in vitro with purified human telomerase reverse transcriptase and human telomerase RNA component. AB - Telomerase is a specialized reverse transcriptase that catalyzes elongation of the telomeric tandem repeat, TTAGGG, by addition of this sequence to the ends of existing telomeres. Human telomerase reverse transcriptase (hTERT) has been identified as a catalytic enzyme involved in telomere elongation that requires telomerase RNA, human telomerase RNA component (hTR), as an RNA template. We established a new method to express and purify soluble insect-expressed recombinant hTERT. The partially purified FLAG-hTERT retained the catalytic activity of telomerase in a complementation assay in vitro to exhibit telomerase activity in telomerase-negative TIG3 cell extract and in a reconstitution assay with FLAG-hTERT and purified hTR in vitro. FLAG-hTERT (D712A) with a mutation in the VDV motif exhibited no telomerase activity, confirming the authentic catalytic activity of FLAG-hTERT. The reconstituted complex of FLAG-hTERT and hTR in vitro was detected by electrophoretic mobility shift assay, and its activity was stimulated by more than 30-fold by TIG3 cell extract. This suggested that some cellular component(s) in the extract facilitated the reconstituted telomerase activity in vitro. Geldanamycin had no effect on the reconstituted activity but partially reduced the stimulated activity of the reconstituted telomerase by the TIG3 cell extract, suggesting that Hsp90 may contribute to the stimulatory effect of the cellular components. PMID- 10811634 TI - Protein substrate binding induces conformational changes in the chaperonin GroEL. A suggested mechanism for unfoldase activity. AB - Chaperonins are molecules that assist proteins during folding and protect them from irreversible aggregation. We studied the chaperonin GroEL and its interaction with the enzyme human carbonic anhydrase II (HCA II), which induces unfolding of the enzyme. We focused on conformational changes that occur in GroEL during formation of the GroEL-HCA II complex. We measured the rate of GroEL cysteine reactivity toward iodo[2-(14)C]acetic acid and found that the cysteines become more accessible during binding of a cysteine free mutant of HCA II. Spin labeling of GroEL with N-(1-oxyl-2,2,5, 5-tetramethyl-3 pyrrolidinyl)iodoacetamide revealed that this additional binding occurred because buried cysteine residues become accessible during HCA II binding. In addition, a GroEL variant labeled with 6-iodoacetamidofluorescein exhibited decreased fluorescence anisotropy upon HCA II binding, which resembles the effect of GroES/ATP binding. Furthermore, by producing cysteine-modified GroEL with the spin label N-(1-oxyl-2,2,5, 5-tetramethyl-3-pyrrolidinyl)iodoacetamide and the fluorescent label 5-((((2-iodoacetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid, we detected increases in spin-label mobility and fluorescence intensity in GroEL upon HCA II binding. Together, these results show that conformational changes occur in the chaperonin as a consequence of protein substrate binding. Together with previous results on the unfoldase activity of GroEL, we suggest that the chaperonin opens up as the substrate protein binds. This opening mechanism may induce stretching of the protein, which would account for reported unfoldase activity of GroEL and might explain how GroEL can actively chaperone proteins larger than HCA II. PMID- 10811635 TI - Transcriptional stimulation by hepatocyte nuclear factor-6. Target-specific recruitment of either CREB-binding protein (CBP) or p300/CBP-associated factor (p/CAF). AB - Transcription factors of the ONECUT class, whose prototype is HNF-6, contain a single cut domain and a divergent homeodomain characterized by a phenylalanine at position 48 and a methionine at position 50. The cut domain is required for DNA binding. The homeodomain is required either for DNA binding or for transcriptional stimulation, depending on the target gene. Transcriptional stimulation by the homeodomain involves the F48M50 dyad. We investigate here how HNF-6 stimulates transcription. We identify transcriptionally active domains of HNF-6 that are conserved among members of the ONECUT class and show that the cut domain of HNF-6 participates to DNA binding and, via a LXXLL motif, to transcriptional stimulation. We also demonstrate that, on a target gene to which HNF-6 binds without requirement for the homeodomain, transcriptional stimulation involves an interaction of HNF-6 with the coactivator CREB-binding protein (CBP). This interaction depends both on the LXXLL motif of the cut domain and on the F48M50 dyad of the homeodomain. On a target gene for which the homeodomain is required for DNA binding, but not for transcriptional stimulation, HNF-6 interacts with the coactivator p300/CBP-associated factor but not with CBP. These data show that a transcription factor can act via different, sequence-specific, mechanisms that combine distinct modes of DNA binding with the use of different coactivators. PMID- 10811636 TI - Phospholemman is a substrate for myotonic dystrophy protein kinase. AB - The genetic abnormality in myotonic muscular dystrophy, multiple CTG repeats lie upstream of a gene that encodes a novel protein kinase, myotonic dystrophy protein kinase (DMPK). Phospholemman (PLM), a major membrane substrate for phosphorylation by protein kinases A and C, induces Cl currents (I(Cl(PLM))) when expressed in Xenopus oocytes. To test the idea that PLM is a substrate for DMPK, we measured in vitro phosphorylation of purified PLM by DMPK. To assess the functional effects of PLM phosphorylation we compared I(Cl(PLM)) in Xenopus oocytes expressing PLM alone to currents in oocytes co-expressing DMPK, and examined the effect of DMPK on oocyte membrane PLM expression. We found that PLM is indeed a good substrate for DMPK in vitro. Co-expression of DMPK with PLM in oocytes resulted in a reduction in I(Cl(PLM)). This was most likely a specific effect of phosphorylation of PLM by DMPK, as the effect was not present in oocytes expressing a phos(-) PLM mutant in which all potential phosphorylation had been disabled by Ser --> Ala substitution. The biophysical characteristics of I(Cl(PLM)) were not changed by DMPK or by the phos(-) mutation. Co-expression of DMPK reduced the expression of PLM in oocyte membranes, suggesting a possible mechanism for the observed reduction in I(Cl(PLM)) amplitude. These data show that PLM is a substrate for phosphorylation by DMPK and provide functional evidence for modulation of PLM function by phosphorylation. PMID- 10811637 TI - Identification of an erythroid active element in the transferrin receptor gene. AB - Hemoglobin synthesis consumes most of the iron that is taken up by cells from plasma transferrin, and this process requires very high expression of transferrin receptors (TfR) at the membranes of erythroid cells. Studies in our and other laboratories indicate that a dramatic increase in TfR levels during erythroid differentiation occurs at the transcriptional level. In this study, we investigated the transcriptional regulation of the TfR in terms of its promoter activity and DNA-protein binding in murine erythroleukemia cells. Reporter gene assays revealed that the TfR promoter activity was stimulated 6-8-fold in murine erythroleukemia cells induced to differentiate into hemoglobin-synthesizing cells by either Me(2)SO or N,N'-hexamethylene-bis-acetamide. A minimal region (-118 to +14) was required for the differentiation-induced promoter activity. Mutation of either an Ets-binding site or an activator protein-1/cyclic AMP-response element like motif within this region, but not disruption of the adjacent GC rich/specificity protein-1 sequence, inhibited the inducible promoter activity. Electrophoresis mobility shift assays suggest that the cyclic AMP-response element-binding proteins/activating transcription factor-like factors and Ets like factors bind constitutively to this bipartite element. Upon induction of differentiation, a shift in the pattern of the cyclic AMP-response element binding protein/activating transcription factor-like binding factors was observed. Our data indicate that the TfR gene promoter contains an erythroid active element that stimulates the receptor gene transcription upon induction of hemoglobin synthesis. PMID- 10811638 TI - Functional consequence of substitutions at residue 171 in human galactose-1 phosphate uridylyltransferase. AB - Impairment of the human enzyme galactose-1-phosphate uridylyltransferase (hGALT) results in the potentially lethal disorder classic galactosemia. Although a variety of naturally occurring mutations have been identified in patient alleles, few have been well characterized. We have explored the functional significance of a common patient mutation, F171S, using a strategy of conservative substitution at the defined residue followed by expression of the wild-type and, alternatively, substituted proteins in a null-background strain of yeast. As expected from patient studies, the F171S-hGALT protein demonstrated <0.1% wild type levels of activity, although two of three conservatively substituted moieties, F171L- and F171Y-hGALT, demonstrated approximately 10% and approximately 4% activity, respectively. The third protein, F171W, demonstrated severely reduced abundance, precluding further study. Detailed kinetic analyses of purified wild-type, F171L- and F171Y-hGALT enzymes, coupled with homology modeling of these proteins, enabled us to suggest that the effects of these substitutions resulted largely from altering the position of a catalytically important residue, Gln-188, and secondarily, by altering the subunit interface and perturbing hexose binding to the uridylylated enzyme. These results not only provide insight into the functional impact of a single common patient allele and offer a paradigm for similar studies of other clinically or biochemically important residues, but they further help to elucidate activity of the wild-type human GALT enzyme. PMID- 10811639 TI - Molecular cloning and expression of mammalian peroxisomal trans-2-enoyl-coenzyme A reductase cDNAs. AB - Chain elongation of fatty acids is an important cellular process and is believed to occur in the endoplasmic reticulum of all eukaroytic cells. Herein we describe the cloning and characterization of a peroxisomal NADPH-specific trans-2-enoyl CoA reductase, the key enzyme for a proposed peroxisomal chain elongation pathway. The reductase was solubilized and partially purified from guinea pig liver peroxisomes by affinity chromatography. On SDS-polyacrylamide gel electrophoresis, a 40-kDa band was identified as the enzyme, and its partial amino acid sequence (27 amino acids) was determined. A full-length cDNA for the reductase was cloned from a guinea pig liver cDNA library. The open reading frame of this nucleotide sequence encodes a 302-amino acid polypeptide with a calculated molecular mass of 32.5 kDa. Full-length mouse and human cDNA clones encoding homologous proteins have also been isolated. All of these translated polypeptides have the type I peroxisomal targeting signal, AKL, at the carboxyl terminus. The identity of the cloned enoyl-CoA reductase cDNAs was confirmed by expressing the guinea pig and human cDNAs in Escherichia coli. The His-tagged recombinant enzymes were found to have very high NADPH-specific 2-enoyl-CoA reductase activity with similar properties and specificity as the liver peroxisomal reductase. Both the natural and the recombinant enzyme catalyze the reduction of trans-2-enoyl-CoAs of varying chain lengths from 6:1 to 16:1, having maximum activity with 10:1 CoA. Northern blot analysis demonstrated that a single transcript of 1.3 kilobases is present in most mouse tissues, with particularly high concentrations in liver and kidney. PMID- 10811640 TI - Human lactoferrin binds and removes the hemoglobin receptor protein of the periodontopathogen Porphyromonas gingivalis. AB - Porphyromonas gingivalis possesses a hemoglobin receptor (HbR) protein on the cell surface as one of the major components of the hemoglobin utilization system in this periodontopathogenic bacterium. HbR is intragenically encoded by the genes of an arginine-specific cysteine proteinase (rgpA), lysine-specific cysteine proteinase (kgp), and a hemagglutinin (hagA). Here, we have demonstrated that human lactoferrin as well as hemoglobin have the abilities to bind purified HbR and the cell surface of P. gingivalis through HbR. The interaction of lactoferrin with HbR led to the release of HbR from the cell surface of P. gingivalis. This lactoferrin-mediated HbR release was inhibited by the cysteine proteinase inhibitors effective to the cysteine proteinases of P. gingivalis. P. gingivalis could not utilize lactoferrin for its growth as an iron source and, in contrast, lactoferrin inhibited the growth of the bacterium in a rich medium containing hemoglobin as the sole iron source. Lactoferricin B, a 25-amino acid long peptide located at the N-lobe of bovine lactoferrin, caused the same effects on P. gingivalis cells as human lactoferrin, indicating that the effects of lactoferrin might be attributable to the lactoferricin region. These results suggest that lactoferrin has a bacteriostatic action on P. gingivalis by binding HbR, removing it from the cell surface, and consequently disrupting the iron uptake system from hemoglobin. PMID- 10811641 TI - Casein kinase I-dependent phosphorylation within a PEST sequence and ubiquitination at nearby lysines signal endocytosis of yeast uracil permease. AB - Uracil uptake by Saccharomyces cerevisiae is mediated by the FUR4-encoded uracil permease. The modification of uracil permease by phosphorylation at the plasma membrane is a key mechanism for regulating endocytosis of this protein. This modification in turn facilitates its ubiquitination and internalization. Following endocytosis, the permease is targeted to the lysosome/vacuole for proteolysis. We have previously shown that uracil permease is phosphorylated at several serine residues within a well characterized N-terminal PEST sequence. In this report, we provide evidence that lysine residues 38 and 41, adjacent to the PEST sequence, are the target sites for ubiquitination of the permease. Conservative substitutions at both Lys(38) and Lys(41) give variant permeases that are phosphorylated but fail to internalize. The PEST sequence contains potential phosphorylation sites conforming to the consensus sequences for casein kinase 1. Casein kinase 1 (CK1) protein kinases, encoded by the redundant YCKI and YCK2 genes, are located at the plasma membrane. Either alone supports growth, but loss of function of both is lethal. Here, we show that in CK1-deficient cells, the permease is poorly phosphorylated and poorly ubiquitinated. Moreover, CK1 overproduction rescued the defective endocytosis of a mutant permease in which the serine phosphoacceptors were replaced by threonine (a less effective phosphoacceptor), which suggests that Yck activity may play a direct role in phosphorylating the permease. Permease internalization was not greatly affected in CK1-deficient cells, despite the low level of ubiquitination of the protein. This may be due to CK1 having a second counteracting role in endocytosis as shown by the higher turnover of variant permeases with unphosphorylatable versions of the PEST sequence. PMID- 10811642 TI - Catalytic roles for two water bridged residues (Asp-98 and His-255) in the active site of copper-containing nitrite reductase. AB - Two active site residues, Asp-98 and His-255, of copper-containing nitrite reductase (NIR) from Alcaligenes faecalis have been mutated to probe the catalytic mechanism. Three mutations at these two sites (D98N, H255D, and H255N) result in large reductions in activity relative to native NIR, suggesting that both residues are involved intimately in the reaction mechanism. Crystal structures of these mutants have been determined using data collected to better than 1. 9-A resolution. In the native structure, His-255 Nepsilon2 forms a hydrogen bond through a bridging water molecule to the side chain of Asp-98, which also forms a hydrogen bond to a water or nitrite oxygen ligated to the active site copper. In the D98N mutant, reorientation of the Asn-98 side chain results in the loss of the hydrogen bond to the copper ligand water, consistent with a negatively charged Asp-98 directing the binding and protonation of nitrite in the native enzyme. An additional solvent molecule is situated between residues 255 and the bridging water in the H255N and H255D mutants and likely inhibits nitrite binding. The interaction of His-255 with the bridging water appears to be necessary for catalysis and may donate a proton to reaction intermediates in addition to Asp-98. PMID- 10811643 TI - Translational control of the antiapoptotic function of Ras. AB - Activated Ras has been shown to provide powerful antiapoptotic signals to cells through well defined transcriptional and post- translational pathways, whereas translational control as a mechanism of Ras survival signaling remains unexplored. Here we show a direct relationship between assembly of the cap dependent translation initiation apparatus and suppression of apoptosis by oncogenic Ras in vitro and in vivo. Decreasing protein synthesis with rapamycin, which is known to inhibit cap-dependent translation, increases the susceptibility of Ras-transformed fibroblasts to cytostatic drug-induced apoptosis. In contrast, suppressing global protein synthesis with equipotent concentrations of cycloheximide actually prevents apoptosis. Enforced expression of the cap dependent translational repressor, the eukaryotic translation initiation factor (eIF) 4E-binding protein (4E-BPI), sensitizes fibroblasts to apoptosis in a manner strictly dependent on its ability to sequester eIF4E from a translationally active complex with eIF4GI and the co-expression of oncogenic Ras. Ectopic expression of 4E-BP1 also promotes apoptosis of Ras-transformed cells injected into immunodeficient mice and markedly diminishes their tumorigenicity. These results establish that eIF4E-dependent protein synthesis is essential for survival of fibroblasts bearing oncogenic Ras and support the concept that activation of cap-dependent translation by extracellular ligands or intrinsic survival signaling molecules suppresses apoptosis, whereas synthesis of proteins mediating apoptosis can occur independently of the cap. PMID- 10811644 TI - Characterization of interleukin-1 receptor-associated kinase in normal and endotoxin-tolerant cells. AB - Interleukin-1 receptor-associated kinase (IRAK), a signal transducer for interleukin-1, has also been suggested to participate in the Toll-like receptor mediated innate immune response to bacterial endotoxin lipopolysaccharide (LPS). Using the human promonocytic THP-1 cell line, we demonstrated that the endogenous IRAK is quickly activated in response to bacterial LPS stimulation, as measured by its in vitro kinase activity toward myelin basic protein. LPS also triggers the association of IRAK with MyD88, the adaptor protein linking IRAK to the Toll like receptor/interleukin-1beta receptor intracellular domain. Macrophage cells with prolonged LPS treatment become tolerant to additional dose of LPS and no longer express inflammatory cytokines. Endotoxin tolerance is a common phenomenon observed in blood from sepsis patients. We observed for the first time that the quantity of IRAK is greatly reduced in LPS-tolerant THP-1 cells, and its activity no longer responds to further LPS challenge. In addition, IRAK does not associate with MyD88 in the tolerant cells. Furthermore, application of AG126, a putative tyrosine kinase inhibitor, can substantially alleviate the LPS-induced cytokine gene expression and can also decrease IRAK level and activity. Our study indicates that IRAK is essential for LPS-mediated signaling and that cells may develop endotoxin tolerance by down-regulating IRAK. PMID- 10811645 TI - Cloning and expression of secretagogin, a novel neuroendocrine- and pancreatic islet of Langerhans-specific Ca2+-binding protein. AB - We have cloned a novel pancreatic beta cell and neuroendocrine cell-specific calcium-binding protein termed secretagogin. The cDNA obtained by immunoscreening a human pancreatic cDNA library using the recently described murine monoclonal antibody D24 contains an open reading frame of 828 base pairs. This codes for a cytoplasmic protein with six putative EF finger hand calcium-binding motifs. The gene could be localized to chromosome 6 by alignment with GenBank genomic sequence data. Northern blot analysis demonstrated abundant expression of this protein in the pancreas and to a lesser extent in the thyroid, adrenal medulla, and cortex. In addition it was expressed in scant quantity in the gastrointestinal tract (stomach, small intestine, and colon). Thyroid tissue expression of secretagogin was restricted to C-cells. Using a sandwich capture enzyme-linked immunosorbent assay with a detection limit of 6.5 pg/ml, considerable amounts of constitutively secreted protein could be measured in tissue culture supernatants of stably transfected RIN-5F and dog insulinoma (INS H1) cell clones; however, in stably transfected Jurkat cells, the protein was only secreted upon CD3 stimulation. Functional analysis of transfected cell lines expressing secretagogin revealed an influence on calcium flux and cell proliferation. In RIN-5F cells, the antiproliferative effect is possibly due to secretagogin-triggered down-regulation of substance P transcription. PMID- 10811646 TI - Nuclear import of insulin-like growth factor-binding protein-3 and -5 is mediated by the importin beta subunit. AB - Although insulin-like growth factor-binding protein (IGFBP)-3 and IGFBP-5 are known to modulate cell growth by reversibly sequestering extracellular insulin like growth factors, several reports have suggested that IGFBP-3, and possibly also IGFBP-5, have important insulin-like growth factor-independent effects on cell growth. These effects may be related to the putative nuclear actions of IGFBP-3 and IGFBP-5, which we have recently shown are transported to the nuclei of T47D breast cancer cells. We now describe the mechanism for nuclear import of IGFBP-3 and IGFBP-5. In digitonin-permeabilized cells, where the nuclear envelope remained intact, nuclear translocation of wild-type IGFBP-3 appears to occur by a nuclear localization sequence (NLS)-dependent pathway mediated principally by the importin beta nuclear transport factor and requiring both ATP and GTP hydrolysis. Under identical conditions, an NLS mutant form of IGFBP-3, IGFBP-3[(228)KGRKR --> MDGEA], was unable to translocate to the nucleus. In cells where both the plasma membrane and nuclear envelope were permeabilized, wild-type IGFBP-3, but not the mutant form, accumulated in the nucleus, implying that the NLS was also involved in mediating binding to nuclear components. By fusing wild-type and mutant forms of NLS sequences (IGFBP-3 [215-232] and IGFBP-5 [201-218]) to the green fluorescent protein, we identified the critical residues of the NLS necessary and sufficient for nuclear accumulation. Using a Western ligand binding assay, wild type IGFBP-3 and IGFBP-5, but not an NLS mutant form of IGFBP-3, were shown to be recognized by importin beta and the alpha/beta heterodimer but only poorly by importin alpha. Together these results suggest that the NLSs within the C terminal domain of IGFBP-3 and IGFBP-5 are required for importin-beta-dependent nuclear uptake and probably also accumulation through mediating binding to nuclear components. PMID- 10811647 TI - Effect of distal cavity mutations on the formation of compound I in catalase peroxidases. AB - Catalase-peroxidases have a predominant catalase activity but differ from monofunctional catalases in exhibiting a substantial peroxidase activity and in having different residues in the heme cavity. We present a kinetic study of the formation of the key intermediate compound I by probing the role of the conserved distal amino acid triad Arg-Trp-His of a recombinant catalase-peroxidase in its reaction with hydrogen peroxide, peroxoacetic acid, and m-chloroperbenzoic acid. Both the wild-type enzyme and six mutants (R119A, R119N, W122F, W122A, H123Q, H123E) have been investigated by steady-state and stopped-flow spectroscopy. The turnover number of catalase activity of R119A is 14.6%, R119N 0.5%, H123E 0.03%, and H123Q 0.02% of wild-type activity. Interestingly, W122F and W122A completely lost their catalase activity but retained their peroxidase activity. Bimolecular rate constants of compound I formation of the wild-type enzyme and the mutants have been determined. The Trp-122 mutants for the first time made it possible to follow the transition of the ferric enzyme to compound I by hydrogen peroxide spectroscopically underlining the important role of Trp-122 in catalase activity. The results demonstrate that the role of the distal His-Arg pair in catalase peroxidases is important in the heterolytic cleavage of hydrogen peroxide (i.e. compound I formation), whereas the distal tryptophan is essential for compound I reduction by hydrogen peroxide. PMID- 10811648 TI - Molecular characterization of the S-adenosyl-L-methionine:3'-hydroxy-N methylcoclaurine 4'-O-methyltransferase involved in isoquinoline alkaloid biosynthesis in Coptis japonica. AB - S-adenosyl-L-methionine:3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4' OMT) catalyzes the conversion of 3'-hydroxy-N-methylcoclaurine to reticuline, an important intermediate in synthesizing isoquinoline alkaloids. In an earlier step in the biosynthetic pathway to reticuline, another O-methyltransferase, S adenosyl-L-methionine:norcoclaurine 6-O-methyltransferase (6-OMT), catalyzes methylation of the 6-hydroxyl group of norcoclaurine. We isolated two kinds of cDNA clones that correspond to the internal amino acid sequences of a 6-OMT/4' OMT preparation from cultured Coptis japonica cells. Heterologously expressed proteins had 6-OMT or 4'-OMT activities, indicative that each cDNA encodes a different enzyme. 4'-OMT was purified using recombinant protein, and its enzymological properties were characterized. It had enzymological characteristics similar to those of 6-OMT; the active enzyme was the dimer of the subunit, no divalent cations were required for activity, and there was inhibition by Fe(2+), Cu(2+), Co(2+), Zn(2+), or Ni(2+), but none by the SH reagent. 4'-OMT clearly had different substrate specificity. It methylated (R,S)-6-O-methylnorlaudanosoline, as well as (R, S)-laudanosoline and (R,S)-norlaudanosoline. Laudanosoline, an N methylated substrate, was a much better substrate for 4'-OMT than norlaudanosoline. 6-OMT methylated norlaudanosoline and laudanosoline equally. Further characterization of the substrate saturation and product inhibition kinetics indicated that 4'-OMT follows an ordered Bi Bi mechanism, whereas 6-OMT follows a Ping-Pong Bi Bi mechanism. The molecular evolution of these two related O-methyltransferases is discussed. PMID- 10811649 TI - Structure of a conserved domain common to the transcription factors TFIIS, elongin A, and CRSP70. AB - TFIIS is a transcription elongation factor that consists of three domains. We have previously solved the structures of domains II and III, which stimulate arrested polymerase II elongation complexes in order to resume transcription. Domain I is conserved in evolution from yeast to human species and is homologous to the transcription factors elongin A and CRSP70. Domain I also interacts with the transcriptionally active RNA polymerase II holoenzyme and therefore, may have a function unrelated to the previously described transcription elongation activity of TFIIS. We have solved the structure of domain I of yeast TFIIS using NMR spectroscopy. Domain I is a compact four-helix bundle that is structurally independent of domains II and III of the TFIIS. Using the yeast structure as a template, we have modeled the homologous domains from elongin A and CRSP70 and identified a conserved positively charged patch on the surface of all three proteins, which may be involved in conserved functional interactions with the transcriptional machinery. PMID- 10811650 TI - Rapid turnover of calcium in the endoplasmic reticulum during signaling. Studies with cameleon calcium indicators. AB - HEK293 cells expressing the thyrotropin-releasing hormone (TRH) receptor were transfected with cameleon Ca(2+) indicators designed to measure the free Ca(2+) concentration in the cytoplasm, [Ca(2+)](cyt), and the endoplasmic reticulum (ER), [Ca(2+)](er). Basal [Ca(2+)](cyt) was about 50 nm; thyrotropin-releasing hormone (TRH) or other agonists increased [Ca(2+)](cyt) to 1 micrometer or higher. Basal [Ca(2+)](er) averaged 500 micrometer and fell to 50-100 micrometer over 10 min in the presence of thapsigargin. TRH consistently decreased [Ca(2+)](er) to 100 micrometer, independent of extracellular Ca(2+), whereas agonists for endogenous receptors generally caused a smaller decline. When added with thapsigargin, all agonists rapidly decreased [Ca(2+)](er) to 5-10 micrometer, indicating that there is substantial store refilling during signaling. TRH increased [Ca(2+)](cyt) and decreased [Ca(2+)](er) if applied after other agonists, whereas other agonists did not alter [Ca(2+)](cyt) or [Ca(2+)](er) if added after TRH. When Ca(2+) was added back to cells that had been incubated with TRH in Ca(2+)-free medium, [Ca(2+)](cyt) and [Ca(2+)](er) increased rapidly. The increase in [Ca(2+)](er) was only partially blocked by thapsigargin but was completely blocked if cells were loaded with 1, 2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. In conclusion, these new Ca(2+) indicators showed that basal [Ca(2+)](er) is approximately 500 micrometer, that [Ca(2+)](er) has to be >100 micrometer to support an increase in [Ca(2+)](cyt) by agonists, and that during signaling, intracellular Ca(2+) stores are continuously refilled with cytoplasmic Ca(2+) by the sarcoendoplasmic reticulum Ca(2+)-ATPase pump. PMID- 10811651 TI - Hysteretic behavior of methionine adenosyltransferase III. Methionine switches between two conformations of the enzyme with different specific activity. AB - Methionine adenosyltransferase III (MATIII) catalyzes S-adenosylmethionine (AdoMet) synthesis and, as part of its reaction mechanism, it also hydrolyzes tripolyphosphate. Tripolyphosphatase activity was linear over time and had a slightly sigmoidal behavior with an affinity in the low micromolar range. On the contrary, AdoMet synthetase activity showed a lag phase that was independent of protein concentration but decreased at increasing substrate concentrations. Tripolyphosphatase activity, which appeared to be slower than AdoMet synthesis, was stimulated by preincubation with ATP and methionine so that it matched AdoMet synthetase activity. This stimulation process, which is probably the origin of the lag phase, represents the slow transition between two conformations of the enzyme that could be distinguished by their different tripolyphosphatase activity and sensitivity to S-nitrosylation. Tripolyphosphatase activity appeared to be the rate-determining reaction in AdoMet synthesis and the one inhibited by S nitrosylation. The methionine concentration necessary to obtain half-maximal stimulation was in the range of physiological methionine fluctuations. Moreover, stimulation of MAT activity by methionine was demonstrated in vivo. We propose that the hysteretic behavior of MATIII, in which methionine induces the transition to a higher specific activity conformation, can be considered as an adaptation to the specific functional requirements of the liver. PMID- 10811652 TI - Interferon-gamma induces secretory group IIA phospholipase A2 in human arterial smooth muscle cells. Involvement of cell differentiation, STAT-3 activation, and modulation by other cytokines. AB - Increased expression of secretory non-pancreatic phospholipase A(2) (sPLA(2)-IIA) could be part of the inflammatory reaction in atherosclerosis. However, the factors controlling sPLA(2)-IIA production in human vascular cells are unknown. We investigated regulation of sPLA(2)-IIA expression and secretion by human arterial smooth muscle cells in culture (HASMC). SPLA(2)-IIA was induced after 3 14 days of culture in non-proliferating conditions. SPLA(2)-IIA was co-expressed with heavy caldesmon, a cytoskeleton protein, and p27, a G(1) cyclin inhibitor, proteins characteristically expressed by differentiated cells. Further incubation with 50-500 units/ml of interferon (IFN)-gamma significantly increased sPLA(2) IIA mRNA and secretion. IFN-gamma-induced sPLA(2)-IIA was found to be active in cell media and associated with cell membrane proteoglycans. IFN-gamma induced sPLA(2)-IIA expression was antagonized by tumor necrosis factor (TNF)-alpha and interleukin (IL)-10. TNF-alpha added individually induced a significant but transient (4 h) increase in sPLA(2)-IIA secretion. IL-10 by itself did not affect sPLA(2)-IIA expression and secretion. IFN-gamma-stimulated sPLA(2)-IIA transcription involved STAT-3 protein. Interestingly, IL-6 but not IFN-gamma up regulated the sPLA(2)-IIA expression in HepG2 cells, thus sPLA(2)-IIA induction by IFN-gamma response appears to be cell specific. In summary, conditions leading to cell differentiation induced sPLA(2)-IIA expression in HASMC and further exposure to IFN-gamma can up-regulate sPLA(2)-IIA transcription and secretion. This IFN-gamma stimulatory effect can be modulated by other cytokines. PMID- 10811653 TI - Drosophila pro-apoptotic Bcl-2/Bax homologue reveals evolutionary conservation of cell death mechanisms. AB - Genetic analysis of programmed cell death in Drosophila reveals many similarities with mammals. Heretofore, a missing link in the fly has been the absence of any Bcl-2/Bax family members, proteins that function in mammals as regulators of mitochondrial cytochrome c release. A Drosophila homologue of the human killer protein Bok (DBok) was identified. The predicted structure of DBok is similar to pore-forming Bcl-2/Bax family members. DBok induces apoptosis in insect and human cells, which is suppressible by anti-apoptotic human Bcl-2 family proteins. A caspase inhibitor suppressed DBok-induced apoptosis but did not prevent DBok induced cell death. Moreover, DBok targets mitochondria and triggers cytochrome c release through a caspase-independent mechanism. These characteristics of DBok reveal evolutionary conservation of cell death mechanisms in flies and humans. PMID- 10811654 TI - Kinetic mechanism of the histone acetyltransferase GCN5 from yeast. AB - The transcriptional coactivator GCN5 from yeast (yGCN5) is a histone acetyltransferase that is essential for activation of target genes. GCN5 is a member of a large family of histone acetyltransferases that are conserved between yeast and humans. To understand the molecular mechanisms of histone/protein acetylation, a detailed kinetic analysis was performed. Bi-substrate kinetic analysis using acetyl-coenzyme A (AcCoA) and an H3 histone synthetic peptide indicated that both substrates must bind to form a ternary complex before catalysis. Product inhibition studies revealed that the product CoA was a competitive inhibitor versus AcCoA. Desulfo-CoA, a dead-end inhibitor, also demonstrated simple competitive inhibition versus AcCoA. Acetylated (Lys14Ac) H3 peptide displayed noncompetitive inhibition against both H3 peptide and AcCoA. These results support a sequential ternary complex (ordered Bi-Bi) kinetic mechanism, where AcCoA binds first, followed by H3 histone. Acetylated (Lys14Ac) H3 product is released first, and CoA is the last product to leave. Also, two methods were developed to measure the binding affinities of AcCoA/CoA for GCN5. Employing the fluorescent CoA analog etheno-CoA (epsilonCoA, 1-N(6)-etheno-CoA), a K(d) for epsilonCoA of 5.1 +/- 1.1 microm was determined by fluorescence anisotropy. This value was similar to the K(d) value of 8.5 +/- 2.6 microm for AcCoA obtained using equilibrium dialysis and to the K(i) (inhibition constant) of 6.7 microm for CoA obtained from steady-state kinetic assays. Together, these data suggest that the acetyl moiety of AcCoA contributes little to the binding energy. PMID- 10811655 TI - Reconstitution of light-independent protochlorophyllide reductase from purified bchl and BchN-BchB subunits. In vitro confirmation of nitrogenase-like features of a bacteriochlorophyll biosynthesis enzyme. AB - Protochlorophyllide reductase catalyzes the reductive formation of chlorophyllide from protochlorophyllide during biosynthesis of chlorophylls and bacteriochlorophylls. The light-independent (dark) form of protochlorophyllide reductase plays a key role in the ability of gymnosperms, algae, and photosynthetic bacteria to green (form chlorophyll) in the dark. Genetic and sequence analyses have indicated that dark protochlorophyllide reductase consists of three protein subunits that exhibit significant sequence similarity to the three subunits of nitrogenase, which catalyzes the reductive formation of ammonia from dinitrogen. However, unlike the well characterized features of nitrogenase, there has been no previous biochemical characterization of dark protochlorophyllide reductase. In this study, we report the first reproducible demonstration of dark protochlorophyllide reductase activity from purified protein subunits that were isolated from the purple nonsulfur photosynthetic bacterium Rhodobacter capsulatus. Two of the three subunits (Bchl and BchN) were expressed in R. capsulatus as S tag fusion proteins that facilitated affinity purification. The third subunit (BchB) was co-purified with the BchN protein indicating that BchN and BchB proteins form a tight complex. Dark protochlorophyllide reductase activity was shown to be dependent on the presence of all three subunits, ATP, and the reductant dithionite. The similarity of dark protochlorophyllide reductase to nitrogenase is discussed. PMID- 10811656 TI - Arginine kinase from Nautilus pompilius, a living fossil. Site-directed mutagenesis studies on the role of amino acid residues in the Guanidino specificity region. AB - Arginine kinases were isolated from the cephalopods Nautilus pompilius, Octopus vulgaris, and Sepioteuthis lessoniana, and the cDNA-derived amino acid sequences have been determined. Although the origin and evolution of cephalopods have long been obscure, this work provides the first molecular evidence for the phylogenetic position of Cephalopoda in molluscan evolution. A crystal structure for Limulus arginine kinase showed that four amino acid residues (Ser(63), Gly(64), Val(65), and Tyr(68)) are hydrogen-bonded with the substrate arginine. We introduced three independent mutations, Ser(63) --> Gly, Ser(63) --> Thr, and Tyr(68) --> Ser, in Nautilus arginine kinase. One of the mutants had a considerably reduced substrate affinity, accompanied by a decreased V(max). In other mutants, the activity was lost almost completely. It is known that substantial conformational changes take place upon substrate binding in arginine kinase. We hypothesize that the hydrogen bond between Asp(62) and Arg(193) stabilizes the closed, substrate-bound state. Site-directed mutagenesis studies strongly support this hypothesis. The mutant (Asp(62) --> Gly or Arg(193) --> Gly), which destabilizes the maintenance of the closed state and/or perhaps disrupts the unique topology of the catalytic pocket, showed only a very weak activity (0.6-1.5% to the wild-type). PMID- 10811657 TI - Intron 1 elements promote erythroid-specific GATA-1 gene expression. AB - The zinc finger protein GATA-1 functions in a concentration-dependent fashion to activate the transcription of erythroid and megakaryocytic genes. Less is understood, however, regarding factors that regulate the GATA-1 gene. Presently elements within intron 1 are shown to markedly affect its erythroid-restricted transcription. Within a full-length 6. 8-kilobase GATA-1 gene construct (G6.8 Luc) the deletion of a central subdomain of intron 1 inhibited transcription >/=10-fold in transiently transfected erythroid SKT6 cells, and likewise inhibited high-level transcription in erythroid FDCW2ER-GATA1 cells. In parental myeloid FDCER cells, however, low-level transcription was largely unaffected by intron 1 deletions. Within intron 1, repeated GATA and Ap1 consensus elements in a central region are described which when linked directly to reporter cassettes promote transcription in erythroid SKT6 and FDCER-GATA1 cells at high rates. Moreover, GATA-1 activated transcription from this subdomain in 293 cells, and in SKT6 cells this subdomain footprinted in vivo. For stably integrated GFP reporter constructs in erythroid SKT6 cells, corroborating results were obtained. Deletion of intronic GATA and Ap1 motifs abrogated the activity of G6.8-pEGFP; activity was decreased by 43 and 56%, respectively, by the deletion of either motif; and the above 1800-base pair region of intron 1 per se was transcribed at rates uniformly greater than G6.8-pEGFP. Also described is the differential utilization of exons 1a and 1b among primary erythromegakaryocytic and myeloid cells. PMID- 10811658 TI - Regulation of Na,K-ATPase beta 1 subunit gene transcription by low external potassium in cardiac myocytes. Role of Sp1 AND Sp3. AB - Expression of Na,K-ATPase activity is up-regulated in cells incubated for extended intervals in the presence of low external K(+). Our previous data showed that exposure of cardiac myocytes to low K(+) increased the steady-state abundance of Na,K-ATPase beta1 subunit mRNA. In the present study we determined that incubation of primary cultures of neonatal rat cardiac myocytes with low K(+) augmented Na,K-ATPase beta1 gene expression at a transcriptional level and that this effect required extracellular Ca(2+). The stimulatory effect of low K(+) on Na,K-ATPase beta1 gene transcription was not dependent on increased contractile activity of cardiac myocytes. Na,K-ATPase beta1 5'-flanking region deletion plasmids used in transient transfection analysis demonstrated that the region between nucleotides -62 to -42 of the beta1 promoter contained a low K(+) response element. Site-directed mutagenesis of a potential GC box core motif GCG in the -58/-56 region of the beta1 promoter decreased basal and low K(+)-mediated transcription. Mutation of the core sequence of a putative GC box element located between nucleotides -101 and -99 further decreased the low K(+) effect on beta1 gene transcription. Electrophoretic mobility shift assays using oligonucleotides spanning the proximal and distal GC box elements of the beta1 promoter showed enhanced binding of two complexes in response to low K(+). The inclusion of a consensus GC box sequence as a competitor in gel shift analysis reduced factor binding to the low K(+) response elements. Antibodies to transcription factors Sp1 and Sp3 interacted with components of both DNA-binding complexes and binding of nuclear factors was abolished in gel shift studies using GC box mutants. Together these data indicate that enhanced binding of Sp1 and Sp3 to two GC box elements in the rat Na,K-ATPase beta1 subunit gene promoter mediates beta1 gene transcription up-regulation in neonatal rat cardiac myocytes exposed to low external K(+). PMID- 10811659 TI - Importance of arginines 63 and 423 in modulating the bile salt-dependent and bile salt-independent hydrolytic activities of rat carboxyl ester lipase. AB - Previous studies using chemical modification approach have shown the importance of arginine residues in bile salt activation of carboxyl ester lipase (CEL) activity. However, the x-ray crystal structure of CEL failed to show the involvement of arginine residues in CEL-bile salt interaction. The current study used a site-specific mutagenesis approach to determine the role of arginine residues 63 and 423 in bile salt-dependent and bile salt-independent hydrolytic activities of rat CEL. Mutations of Arg(63) to Ala(63) (R63A) and Arg(423) to Gly(423) (R423G) resulted in enzymes with increased bile salt-independent hydrolytic activity against lysophosphatidylcholine, having 6.5- and 2-fold higher k(cat) values, respectively, in comparison to wild type CEL. In contrast, the R63A and R423A mutant enzymes displayed 5- and 11-fold decreases in k(cat), in comparison with wild type CEL, for bile salt-dependent cholesteryl ester hydrolysis. Although taurocholate induced similar changes in circular dichroism spectra for wild type, R63A, and R423G proteins, this bile salt was less efficient in protecting the mutant enzymes against thermal inactivation in comparison with control CEL. Lipid binding studies revealed less R63A and R423G mutant CEL were bound to 1,2-diolein monolayer at saturation compared with wild type CEL. These results, along with computer modeling of the CEL protein, indicated that Arg(63) and Arg(423) are not involved directly with monomeric bile salt binding. However, these residues participate in micellar bile salt modulation of CEL enzymatic activity through intramolecular hydrogen bonding with the C-terminal domain. These residues are also important, probably through similar intramolecular hydrogen bond formation, in stabilizing the enzyme in solution and at the lipid-water interface. PMID- 10811660 TI - Crystal structure and activity of human p23, a heat shock protein 90 co chaperone. AB - p23 is a co-chaperone for the heat shock protein, hsp90. This protein binds hsp90 and participates in the folding of a number of cell regulatory proteins, but its activities are still unclear. We have solved a crystal structure of human p23 lacking 35 residues at the COOH terminus. The structure reveals a disulfide linked dimer with each subunit containing eight beta-strands in a compact antiparallel beta-sandwich fold. In solution, however, p23 is primarily monomeric and the dimer appears to be a minor component. Conserved residues are clustered on one face of the monomer and define a putative surface region and binding pocket for interaction(s) with hsp90 or protein substrates. p23 contains a COOH terminal tail that is apparently less structured and is unresolved in the crystal structure. This tail is not needed for the binding of p23 to hsp90 or to complexes with the progesterone receptor. However, the tail is necessary for optimum active chaperoning of the progesterone receptor, as well as the passive chaperoning activity of p23 in assays measuring inhibition of heat-induced protein aggregation. PMID- 10811661 TI - Phosphorylation of human gp130 at Ser-782 adjacent to the Di-leucine internalization motif. Effects on expression and signaling. AB - The receptor for leukemia inhibitory factor (LIF) consists of two polypeptides, the LIF receptor and gp130. Agonist stimulation has been shown previously to cause phosphorylation of gp130 on serine, threonine, and tyrosine residues. We found that gp130 fusion proteins were phosphorylated exclusively on Ser-782 by LIF- and growth factor-stimulated 3T3-L1 cell extracts. Ser-780 was required for phosphorylation of Ser-782 but was not itself phosphorylated. Ser-782 is located immediately N-terminal to the di-leucine motif of gp130, which regulates internalization of the receptor. Transient expression of chimeric granulocyte colony-stimulating factor receptor (G-CSFR)-gp130(S782A) receptors resulted in increased cell surface expression in COS-7 cells and increased ability to induce vasoactive intestinal peptide gene expression in IMR-32 neuroblastoma cells when compared with expression of chimeric receptors containing wild-type gp130 cytoplasmic domains. These results identify Ser-782 as the major phosphorylated serine residue in human gp130 and indicate that this site regulates cell surface expression of the receptor polypeptide. PMID- 10811662 TI - The p53/retinoblastoma-mediated repression of testicular orphan receptor-2 in the rhesus monkey with cryptorchidism. AB - Whereas the linkage of infertility to cryptorchidism, the failure of the testis to descend into the scrotum at birth, has been well documented, the detailed molecular mechanism remains unclear. Here we report that the testicular orphan receptor-2 (TR2) expression, which modulates many signal pathways, was completely repressed in the surgery-induced cryptorchidism of the rhesus monkey. Further studies link TR2 repression to the induction of p53 and results suggest that induced p53 could repress TR2 expression via the p53-->p21-->CDK-->Rb-->E2F signal pathway. In return, TR2 could also control the expression of p53 and Rb through the regulation of human papillomavirus 16 E6/E7 genes. Together, our data suggest a feedback control mechanism between TR2 and p53/Rb tumor suppressors, which might play important roles in male infertility associated with cryptorchidism. PMID- 10811663 TI - The curious case of the baffling biological. PMID- 10811664 TI - Interferon alpha consolidation after intensive chemotherapy does not prolong the progression-free survival of patients with low-grade non-Hodgkin's lymphoma: results of the Southwest Oncology Group randomized phase III study 8809. AB - PURPOSE: S8809 is a randomized phase III trial determining whether intensive cytoreductive treatment, followed by interferon consolidation at the time of minimal residual disease, prolongs the progression-free survival (PFS) or overall survival (OS) of indolent lymphoma patients. PATIENTS AND METHODS: Five hundred seventy-one patients with previously untreated stage III or IV low-grade non Hodgkin's lymphoma were registered. Patients received six to eight cycles of prednisone, methotrexate, doxorubicin, cyclophosphamide, and etoposide/mechlorethamine, vincristine, procarbazine, and prednisone (ProMACE[day 1]-MOPP[day 8]) chemotherapy or chemotherapy plus radiotherapy. Responding patients were randomized to observation alone or to interferon consolidation. Interferon alpha-2b 2 mU/m(2) was given subcutaneously three times weekly for 2 years. RESULTS: Two hundred sixty-eight eligible patients were randomized to interferon alpha consolidation (n = 144) or observation alone (n = 124). With a median follow-up time from randomization among patients still alive of 6.2 years, the median PFS time was 4.1 years for patients who received interferon consolidation therapy and 3.2 years for patients who were observed after ProMACE MOPP induction (P =.25). The adjusted hazard ratio for relapse for observation to interferon was 0.83 (95% confidence interval [CI], 0.61 to 1.13). The median OS has not been reached in either group. At 5 years, OS is 78% for the interferon group and 77% for the observation group (P =.65). The adjusted hazard ratio for survival for observation to interferon is 1.11 (95% CI, 0.69 to 1. 79). CONCLUSION: Interferon alpha consolidation therapy after intensive treatment with anthracycline-containing combination chemotherapy and involved-field radiation therapy does not prolong the PFS or OS of patients with low-grade non-Hodgkin's lymphoma. PMID- 10811665 TI - Outcome of CNS disease at diagnosis in disseminated small noncleaved-cell lymphoma and B-cell leukemia: a Children's Cancer Group study. AB - PURPOSE: To examine the impact of initial CNS involvement on outcome and patterns of failure in patients with disseminated small noncleaved-cell lymphoma and B cell leukemia who were treated in four successive Children's Cancer Group trials. PATIENTS AND METHODS: Of 462 patients with disseminated disease, 49 (10.6%) had CNS disease at diagnosis (CNS+). CNS disease included meningeal disease or CNS parenchymal masses with or without cranial neuropathies (CSF+/Mass; CNPs) in 36 patients and isolated CNPs in 13. Of the CNS+ patients, 28 had M2 (5% to 25% blasts) or M3 (> 25% blasts) bone marrow involvement. All patients received protocol-based systemic and intrathecal chemotherapy. Thirty-six patients also received CNS irradiation. RESULTS: Relapses occurred in 21 (43%) of 49 patients, predominantly in the CNS (71%) and bone marrow (52%). The 3-year event-free survival +/- SE for all patients with CNS+ disease was 45% +/- 7%. Patients with CSF+/Mass had a nominally higher treatment failure rate compared with patients with CNS- after adjusting for marrow status and lactate dehydrogenase (LDH) diagnosis, with a relative failure rate (RFR) of 1.52 (95% confidence interval [CI], 0.88 to 2.6; P =.15). In comparison, the RFRs for patients with M2 or M3 marrow and for those with LDH levels greater than 500 IU/L after adjusting for CNS disease were 1.4 (95% CI, 0.96 to 2.0; P =.029) and 2.2 (95% CI, 1.5 to 3.0; P <.001), respectively. The RFR for patients with isolated CNPs was 0.87 (95% CI, 0.36 to 2.1; P =.76). CONCLUSION: We conclude that, with the treatments used during the period covered by these studies, the presence of CSF+/Mass CNS disease at diagnosis was associated with a nominally worse outcome independent of initial bone marrow status and LDH level, but the effect was not statistically significant. PMID- 10811666 TI - Characteristics and outcome of children with Beckwith-Wiedemann syndrome and Wilms' tumor: a report from the National Wilms Tumor Study Group. AB - PURPOSE: Children with Beckwith-Wiedemann syndrome (BWS) are at increased risk for developing Wilms' tumor (WT). We reviewed the National Wilms Tumor Study Group (NWTSG) records to assess clinical characteristics and outcome of patients with WT and BWS. METHODS: In the NWTSG, treating clinicians were asked to report, for each enrolled patient, whether the patient had BWS. Between 1980 and 1995, 4,669 patients were treated on two consecutive NWTSG protocols (NWTS 3 and NWTS 4). We retrospectively reviewed the clinical characteristics and treatment outcomes of BWS patients compared with patients with WT without BWS. RESULTS: Fifty-three children enrolled onto NWTS 3 and 4 were reported to have BWS. BWS patients were more likely to present with lower-stage tumors (P =.0001), with more than half (27 of 53) presenting with stage I disease. The overall treatment outcomes for the BWS patients were nearly identical to those without BWS, with overall survival at 4 years from diagnosis at 89% and 90%, respectively. Overall, 21% of the patients with BWS had bilateral disease, either at diagnosis (nine of 53) or as metachronous contralateral recurrence (two of 53). BWS patients enrolled onto NWTS 4 had smaller tumors than those enrolled onto NWTS 3 (P =.02), a trend not seen in the non-BWS patients. CONCLUSION: Like children without BWS, children with BWS and WT have an excellent prognosis with modern treatment regimens. There is a high risk of bilateral disease, and increasingly smaller tumors are being detected. This suggests that a national trial assessing the role of ultrasound screening followed by nephron-sparing surgery for some patients may be appropriate. PMID- 10811667 TI - Does paclitaxel improve the chemoradiotherapy of locoregionally advanced esophageal cancer? A nonrandomized comparison with fluorouracil-based therapy. AB - PURPOSE: A phase II trial of accelerated fractionation radiation with concurrent cisplatin and paclitaxel chemotherapy was performed to investigate the role of the paclitaxel, when substituted for fluorouracil (5-FU), in the chemoradiotherapy of esophageal cancer. PATIENTS AND METHODS: Patients with an esophageal ultrasound stage of T(3) or N(1) or M(1) (nodal) esophageal cancer were treated with two courses of a cisplatin infusion (20 mg/m(2)/d for 4 days) and paclitaxel (175 mg/m(2) over 24 hours) concurrent with a split course of accelerated fractionation radiation (1.5 Gy bid to a total dose of 45 Gy). Surgical resection was performed 4 to 6 weeks later followed by a single identical postoperative course of chemoradiotherapy (24 Gy) in patients with significant residual tumor at surgery. Toxicity and results of this treatment were retrospectively compared with our previous 5-FU and cisplatin chemoradiotherapy experience. RESULTS: Between September 1995 and July 1997, 40 patients were entered onto this study. Although dysphagia proved worse in our 5 FU-treated patients, profound leukopenia and a need for unplanned hospitalization were significantly more common in the paclitaxel group. Thirty-seven patients (93%) proved resectable for cure. The 3-year projected overall survival is 30%, locoregional control is 81%, and distant metastatic disease control is 44%. When compared with a similarly staged cohort of 5-FU-treated patients, there was no advantage for any survival function studied. CONCLUSION: This paclitaxel-based treatment regimen for locoregionally advanced esophageal cancer produced increased toxicity with no improvement in results when compared with our previous 5-FU experience. Paclitaxel-based treatments must be carefully and prospectively studied before their incorporation into the standard management of esophageal cancer. PMID- 10811668 TI - Concomitant radiotherapy and chemotherapy for early-stage nasopharyngeal carcinoma. AB - PURPOSE: Early-stage nasopharyngeal carcinoma (NPC) continues to carry a failure rate of 15% to 30% when treated with radiotherapy alone; the benefit of concomitant radiotherapy and chemotherapy (CCRT) in early-stage NPC is unclear. The purpose of this report is to describe our efforts to improve treatment outcome in early-stage NPC after CCRT. PATIENTS AND METHODS: Of 189 newly diagnosed NPC patients without evidence of distant metastases who were treated in our institution between 1990 and 1997, 44 presented with early-stage (stage I and II) disease according to the American Joint Committee on Cancer (AJCC) 1997 NPC staging system. Twelve of these patients were treated with radiotherapy alone and 32 with CCRT. Each patient's head and neck area was evaluated by magnetic resonance imaging or computed tomography. Radiotherapy was administered at 2 Gy per fraction per day, Monday through Friday, for 35 fractions for a total dose of 70 Gy. Chemotherapy consisting of cis-diamine-dichloroplatinum and fluorouracil was delivered simultaneously with radiotherapy in weeks 1 and 6 and sequentially for two monthly cycles after radiotherapy. RESULTS: Patients who were treated with radiotherapy alone primarily had stage I disease, whereas none of those who were treated with CCRT had stage I disease (11 of 12 patients v none of 32 patients; P =.001). The locoregional control rate at 3 years for the radiotherapy group was 91.7% (median follow-up period, 34 months) and was 100% for the CCRT group (median follow-up period, 44 months) (P =.10). The 3-year disease-free survival rate in the radiotherapy group was 91.7% and was 96.9% in the CCRT group (P =.66). CONCLUSION: Our results reveal excellent prognosis of AJCC 1997 stage II NPC treated with CCRT. Stage II patients with a greater tumor burden treated with CCRT showed an equal disease-free survival, compared with stage I patients treated with radiotherapy alone. A prospective randomized trial is underway to confirm the role of CCRT in stage II NPC. PMID- 10811669 TI - Prognostic significance of vascular endothelial growth factor protein levels in oral and oropharyngeal squamous cell carcinoma. AB - PURPOSE: Vascular Endothelial Growth Factor (VEGF) promotes angiogenesis in many different tumor types. VEGF levels may affect tumor growth, metastatic potential, and response to radiotherapy. This study assesses the prognostic value of VEGF protein levels in a cohort of patients with oral and oropharyngeal squamous cell carcinomas. The relationships between clinical outcome and the covariables of tumor-node-metastasis stage, disease stage (I to IV), grade, margin status, race, sex, and age were also determined. PATIENTS AND METHODS: Chart review identified 77 patients with oral or oropharyngeal squamous cell carcinoma treated with gross total surgical resection and postoperative radiation between 1981 and 1992. Sufficient follow-up data and tumor tissue were available in 56 patients (73%). VEGF protein levels were determined using immunohistochemistry. The association between VEGF status, covariables, and outcome was assessed in a bivariate and multivariate model using two-sided statistical tests. RESULTS: Twenty-three tumors (41%) were positive for VEGF expression. VEGF-positive tumors were more likely to recur locally (relative risk [RR] = 3.08; 95% confidence interval [CI], 1.03 to 9.24) and distantly (RR = 4.62; 95% CI, 1.41 to 15.10). In bivariate analysis, VEGF positivity was the most significant predictor of poor disease-free survival (RR = 2.66; 95% CI, 1.27 to 5.56) and overall survival (RR = 3.21; 95% CI, 1.63 to 6.32). In multivariate analysis, VEGF positivity was the most significant predictor of poor disease-free survival (RR = 2.75; 95% CI, 1.30 to 5.79) and overall survival (RR = 3.53; 95% CI, 1.75 to 7.13). CONCLUSION: In this cohort, VEGF positivity was the most significant predictor of poor prognosis. VEGF status may prove to be an important prognostic factor in head and neck cancer. PMID- 10811670 TI - Treatment of leptomeningeal metastases evaluated by interphase cytogenetics. AB - PURPOSE: Although cytologic examination of CSF is the primary method for the evaluation of response to therapy for leptomeningeal metastases (LMMs), the procedure's sensitivity decreases throughout the course of protracted therapy. We studied whether this response could be monitored more accurately through the detection of numerical chromosomal aberrations by interphase cytogenetics, using fluorescence in situ hybridization (FISH). PATIENTS AND METHODS: Seven patients treated for LMMs and with a known numerical aberration for chromosome 1 in their pretreatment CSF were included in this study. Up to 16 consecutive CSF samples were analyzed by means of the fluorescence in situ hybridization (FISH) technique for cells with aberrant chromosome 1 content. The results of routine cytology and FISH analyses were compared and were correlated with each patient's neurologic status. RESULTS: Routine cytology detected malignancies in only 24 of the 76 samples, all of which were classified as chromosomally abnormal by FISH (except for two samples that could not be evaluated). Moreover, FISH demonstrated aneusomic cells in 32 additional samples, which could therefore be classified as malignant. The FISH results correlated better with patient neurologic status in that more malignant cells were detected in the CSF of neurologically deteriorating patients. CONCLUSION: Using FISH in addition to performing routine cytologic examination of CSF led to a more accurate evaluation of response to treatment in patients treated for LMMs. PMID- 10811671 TI - Prognostic value of histologic grade and proliferative activity in axillary node positive breast cancer: results from the Eastern Cooperative Oncology Group Companion Study, EST 4189. AB - PURPOSE: The identification of a subset of patients with axillary lymph node positive breast cancer with an improved prognosis would be clinically useful. We report the prognostic importance of histologic grading and proliferative activity in a cohort of patients with axillary lymph node-positive breast cancer and compare these parameters with other established prognostic factors. PATIENTS AND METHODS: This Eastern Cooperative Oncology Group laboratory companion study (E4189) centered on 560 axillary lymph node-positive patients registered onto one of six eligible clinical protocols. Flow cytometric (ploidy and S-phase fraction [SPF]) and histopathologic analyses (Nottingham Combined Histologic Grade and mitotic index) were performed on paraffin-embedded tissue from 368 patients. RESULTS: Disease recurred in 208 patients; in 161 (77%), within the first 5 years. Mitotic index and grade were associated with both ploidy and SPF (P or = 10 involved axillary nodes, or inflammatory carcinoma. All patients received the same HDCT regimen, with cyclophosphamide, cisplatin, and carmustine (STAMP-I), followed by autologous stem-cell transplantation. Median follow-up was 42 months (range, 5 to 90 months). The same pathologist, blinded to clinical outcome, reviewed all immunostained slides. RESULTS: Positive results for Her-2/neu and p53 were found in 44.5% and 34% of the patients, respectively. Positivity for Her-2/neu was significantly associated with increased risk of relapse and death. No correlation was found between p53 mutations and relapse-free survival (RFS) or overall survival (OS). Multivariate analyses included Her-2/neu overexpression and the following variables previously identified as independent predictors of outcome in this population: tumor size, nodal ratio (number of involved nodes/number of dissected nodes), and hormone receptor status. All four variables had independent value. CONCLUSION: Her-2/neu overexpression is an independent negative predictor of RFS and OS in HRPBC treated with HDCT. Its inclusion in our previously described predictive model increases the predictive capacity of this model for the low-risk subgroup. In contrast, p53 mutations lack predictive value in this setting. PMID- 10811673 TI - Randomized phase II study of docetaxel versus doxorubicin in first- and second line chemotherapy for locally advanced or metastatic soft tissue sarcomas in adults: a study of the european organization for research and treatment of cancer soft tissue and bone sarcoma group. AB - PURPOSE: To assess antitumor response and time to progression (TTP) with docetaxel compared with doxorubicin in first-line treatment of advanced and/or metastatic soft tissue sarcoma. PATIENTS AND METHODS: Patients with measurable soft tissue sarcoma lesions and adequate bone marrow, liver, and renal function were entered onto the study. They were randomized to either docetaxel 100 mg/m(2) given as a 1-hour intravenous infusion every 3 weeks or doxorubicin 75 mg/m(2) given as a bolus injection every 3 weeks. A maximum of seven cycles of treatment were scheduled. The study was designed as a randomized phase III study evaluating TTP by log-rank model. There was a clause for premature closure of the trial if fewer than five responses were observed among the first 25 assessable patients in the docetaxel treatment arm. RESULTS: Eighty-six patients were entered onto the study; 85 were assessable for toxicity and 83 for response. The rate of severe granulocytopenia was not significantly different between the two arms. Nausea (P =.001), vomiting (P <.001), and stomatitis (P =.005) were more common with doxorubicin therapy, whereas neurotoxicity was more frequent with docetaxel treatment. The response rate to doxorubicin therapy was 30% (95% confidence interval, 17% to 46%), whereas no responses to docetaxel therapy were seen (P <.001). In view of this, the trial was closed prematurely and the phase III study part was not conducted. CONCLUSION: Docetaxel is inactive in soft tissue sarcomas and cannot be recommended for further use in treatment of this disease. PMID- 10811674 TI - Synovial sarcoma: a multivariate analysis of prognostic factors in 112 patients with primary localized tumors of the extremity. AB - PURPOSE: Synovial sarcoma is a high-grade tumor that is associated with poor prognosis. Previous studies analyzing prognostic factors are limited because of inclusion of heterogeneous cohorts of patients with nonextremity and recurrent tumors. The objective of this study was to determine independent prognostic factors of primary synovial sarcoma localized to the extremity. PATIENTS AND METHODS: Between July 1, 1982, and June 30, 1996, 112 patients underwent surgical resection for cure at our institution and then were followed-up prospectively. Clinical and pathologic factors examined for prognostic value included age, sex, tumor site and location, depth, size, microscopic status of surgical margins, invasion of bone or neurovascular structures, and monophasic or biphasic histology. The end points analyzed were the time to first local recurrence that was not preceded by a distant recurrence, time to any distant recurrence, and time to disease-related mortality. These end points were modeled using the method of Kaplan and Meier and analyzed by the log-rank test and Cox regression. RESULTS: The median duration of follow-up among survivors in this cohort of 112 patients was 72 months. The 5-year local-recurrence, distant-recurrence, and mortality rates were 12%, 39%, and 25%, respectively. Tumor size > or = 5 cm (P =.001; relative risk [RR] = 2. 7; 95% confidence interval [CI], 1.5 to 5.2) and the presence of bone or neurovascular invasion (P =.04; RR = 2.3; 95% CI, 1.0 to 5. 3) were independent adverse predictors of distant recurrence. Tumor size > or= 5 cm (P =.003; RR = 2.3; 95% CI, 1.4 to 6.3) and the presence of bone or neurovascular invasion (P =.03; RR = 2.7; 95% CI, 1.0 to 6.5) were also independent adverse predictors of mortality. CONCLUSION: The natural history of primary synovial sarcoma of the extremity is related to tumor size and invasion of bone and neurovascular structures. PMID- 10811675 TI - Prospective randomized trial of docetaxel versus best supportive care in patients with non-small-cell lung cancer previously treated with platinum-based chemotherapy. AB - PURPOSE: To evaluate whether treatment with single-agent docetaxel would result in longer survival than would best supportive care in patients with non-small cell lung cancer who had previously been treated with platinum-based chemotherapy. Secondary end points included assessment of response (docetaxel arm only), toxicity, and quality of life. PATIENTS AND METHODS: Patients with performance statuses of 0 to 2 and stage IIIB/IV non-small-cell lung cancer with either measurable or evaluable lesions were eligible for entry onto the study if they had undergone one or more platinum-based chemotherapy regimens and if they had adequate hematology and biochemistry parameters. They were excluded if they had symptomatic brain metastases or if they had previously been treated with paclitaxel. Patients were stratified by performance status and best response to cisplatin chemotherapy and were then randomized to treatment with docetaxel 100 mg/m(2) (49 patients) or 75 mg/m(2) (55 patients) or best supportive care. Patients in both arms were assessed every 3 weeks. RESULTS: One hundred four patients (103 of whom were eligible for entry onto the study) were well balanced for prognostic factors. Of 84 patients with measurable lesions, six (7. 1%) achieved partial responses (three patients at each dose level). Time to progression was longer for docetaxel patients than for best supportive care patients (10.6 v 6.7 weeks, respectively; P <.001), as was median survival (7.0 v 4.6 months; log-rank test, P =.047). The difference was more significant for docetaxel 75 mg/m(2) patients, compared with corresponding best supportive care patients (7.5 v 4.6 months; log-rank test, P =.010; 1-year survival, 37% v 11%; chi(2) test, P =.003). Febrile neutropenia occurred in 11 patients treated with docetaxel 100 mg/m(2), three of whom died, and in one patient treated with docetaxel 75 mg/m(2). Grade 3 or 4 nonhematologic toxicity, with the exception of diarrhea, occurred at a similar rate in both the docetaxel and best supportive care groups. CONCLUSION: Treatment with docetaxel is associated with significant prolongation of survival, and at a dose of 75 mg/m(2), the benefits of docetaxel therapy outweigh the risks. PMID- 10811676 TI - Phase I pharmacologic study of oral topotecan and intravenous cisplatin: sequence dependent hematologic side effects. AB - PURPOSE: In in vitro studies, synergism and sequence-dependent effects were reported for the combination of topotecan and cisplatin. Recently, an oral formulation of topotecan became available. This phase I study was performed to assess the feasibility of the combination of oral topotecan and cisplatin, the pharmacokinetic interaction, and sequence-dependent effects. PATIENTS AND METHODS: Topotecan was administered orally (PO) daily for 5 days in escalating doses and cisplatin was given intravenously (IV) at a fixed dose of 75 mg/m(2) either before topotecan administration on day 1 (sequence CT) or after topotecan administration on day 5 (sequence TC) once every 3 weeks. Patients were treated in a randomized cross-over design. RESULTS: Forty-nine patients were entered onto the study; one patient was not eligible. Sequence CT induced significantly more severe myelosuppression than did sequence TC, and the maximum-tolerated dosage of topotecan in sequence CT was 1.25 mg/m(2)/d x 5. In sequence TC, the maximum tolerated dosage of topotecan was 2.0 mg/m(2)/d x 5. Dose-limiting toxicity consisted of myelosuppression and diarrhea. Pharmacokinetics of topotecan and cisplatin were linear over the dose range studied; no sequence-dependent effects were observed. In addition, topotecan did not influence the protein binding of cisplatin or the platinum-DNA adduct formation in peripheral leukocytes in either sequence. CONCLUSION: The recommended dosages for phase II studies involving patients like the patients in our study are topotecan 1.25 mg/m(2)/d PO x 5 preceded by cisplatin 75 mg/m(2) IV day 1 once every 3 weeks, and topotecan 2.0 mg/m(2)/d PO followed by cisplatin 75 mg/m(2) IV day 5. No pharmacokinetic interaction could be discerned in our study. The antitumor efficacy of both schedules should be evaluated in a randomized phase II study. PMID- 10811677 TI - Sequence effect of epirubicin and paclitaxel treatment on pharmacokinetics and toxicity. AB - PURPOSE: Sequence-dependent clinical and pharmacokinetic interactions between paclitaxel and doxorubicin have been reported. Some data have shown an influence of paclitaxel on epirubicin metabolism, but no data are available about the effect of diverse sequences of these drugs. We investigated whether reversing the sequence of epirubicin and paclitaxel affects the pattern or degree of toxicity and pharmacokinetics. PATIENTS AND METHODS: Patients receiving epirubicin 90 mg/m(2) by intravenous bolus followed by paclitaxel 175 mg/m(2) over 3-hour infusion or the opposite sequence every 3 weeks for four cycles were eligible. Toxicity was recorded at nadir. Pharmacokinetic data were evaluated at the first and the second cycle and were correlated with toxicity parameters. RESULTS: Thirty-nine consecutive stage II breast cancer patients were treated. Twenty-one patients received epirubicin followed by paclitaxel (ET group), and 18 received the opposite sequence (TE group). No significant difference in nonhematologic toxicity was seen. A lower neutrophil and platelet nadir and a statistically significant slower neutrophil recovery was observed in the TE group. Area under the concentration-time curve (AUC) of epirubicin was higher in the TE group (2,346 ng/mL. h v 1,717 ng/mL. h; P =.002). An inverse linear correlation between epirubicin AUC and neutrophil recovery was also observed (P =.012). No difference was detected in paclitaxel pharmacokinetics. CONCLUSION: Our results support a sequence-dependent effect of paclitaxel over epirubicin pharmacokinetics that is associated with increased myelotoxicity. Because schedule modifications of anthracyclines and paclitaxel can have clinical consequences, the classical way of administration (ie, anthracyclines followed by paclitaxel) should be maintained in clinical practice. PMID- 10811678 TI - Rifampin does not improve the efficacy of quinolone antibacterial prophylaxis in neutropenic cancer patients: results of a randomized clinical trial. AB - PURPOSE: To determine whether the addition of rifampin to a quinolone-based antibacterial prophylactic regimen in patients undergoing high-dose chemotherapy (HDC) with peripheral-blood stem-cell transplantation (PBSCT) decreases the incidence of neutropenia and fever, Gram-positive bacteremia, and infection related morbidity. PATIENTS AND METHODS: Patients with solid tumors undergoing HDC with PBSCT were randomized to receive prophylactic antibiotics with either ciprofloxacin 500 mg orally every 8 hours or the same ciprofloxacin regimen with rifampin 300 mg orally every 12 hours. Prophylaxis was started 48 hours before stem-cell reinfusion. Patients were monitored to document the occurrence of neutropenia and fever, incidence and cause of bacterial infection, time to onset and duration of fever, requirement for intravenous antimicrobials, and length of hospital admission. RESULTS: Sixty-five patients were randomized to receive ciprofloxacin and 65 to receive ciprofloxacin plus rifampin, and from these groups, 62 and 61 were assessable, respectively. The proportion of patients who developed neutropenia and fever was 87% in the group treated with ciprofloxacin and 78% in the group treated with ciprofloxacin and rifampin (P =.25). Although there was a trend toward a reduction in the overall incidence of bacteremia (12 v 4 patients), and Gram-positive bacteremia (8 v 2 patients) with the addition of rifampin, none of these comparisons was statistically significant (P =.05 and P =.09, respectively). CONCLUSION: The results of this study, which demonstrate that rifampin does not improve ciprofloxacin antibacterial prophylaxis in cancer patients undergoing HDC with PBSCT support but that it does increase the occurrence of undesirable side effects, do not support the routine use of rifampin in this setting. PMID- 10811679 TI - Anticipated versus actual emotional reactions to disclosure of results of genetic tests for cancer susceptibility: findings from p53 and BRCA1 testing programs. AB - PURPOSE: We examined the ability of individuals undergoing genetic testing for cancer susceptibility in two structured research protocols to accurately anticipate emotional reactions to disclosure of their test result. We explored whether accuracy of emotional anticipation was associated with postdisclosure psychologic adjustment. METHODS: Data from 65 individuals were analyzed; 24 members of Li-Fraumeni cancer syndrome families were tested for p53 mutations (all 24 were unaffected), and 41 subjects with hereditary breast-ovarian cancer susceptibility were tested for BRCA1 mutations (34 were unaffected and seven were affected). Subjects were from families in which a germline mutation had been previously identified. At the pretest session, subjects rated the extent to which they anticipated feeling each of six emotional states (relief, happiness, sadness, guilt, anger, and worry) after disclosure that they did or did not carry the familial mutation. After receiving their test result, they rated their feelings on the same scale of emotions for the appropriate condition. Extent of accuracy and association with psychologic distress at 6 months, as assessed with standardized measures, were evaluated. RESULTS: Overall, mean levels of emotional reactions after receiving test results were not different from those anticipated before result disclosure. However, affected BRCA1 carriers experienced higher levels of anger and worry than they had anticipated. Underestimation of subsequent distress emotions related to test result was associated with a significant increase in general psychologic distress at 6 months. CONCLUSION: Unaffected individuals in cancer-predisposition testing programs are generally accurate in anticipating emotional reactions to test results. However, cancer patients may underestimate their distress after disclosure of positive results and could benefit from intervention strategies. PMID- 10811680 TI - Early depressive symptoms in cancer patients receiving interleukin 2 and/or interferon alfa-2b therapy. AB - PURPOSE: Depressive symptomatology is frequently associated with interleukin (IL) 2 and interferon alfa-2b (INFalpha-2b) therapy in cancer patients. The objective of the present study was to evaluate the depressive and anxiety symptoms induced by IL-2 and/or INFalpha-2b in cancer patients during the first days of cytokine immunotherapy. PATIENTS AND METHODS: The study included 48 patients with renal cell carcinoma or melanoma. Patients were treated either with subcutaneous IL-2, alone (n = 20) or in combination with INFalpha-2b (n = 6); or with INFalpha-2b alone, administered subcutaneously at a low dose (n = 8) or intravenously at a high dose (n = 14). Depressive symptoms were evaluated using the Montgomery and Asberg Depression Rating Scale (MADRS), and anxiety symptoms were evaluated using the Covi scale. Evaluations were performed just before initiation of treatment (day 1) and on days 3 and 5 of treatment. RESULTS: Patients treated with IL-2 alone or in association with INFalpha-2b had significantly higher MADRS scores after 5 days of cytokine therapy, and patients who received both cytokines had increased scores on day 3. In contrast, patients treated with INFalpha-2b alone did not have varying MADRS scores during the course of treatment. Cytokine therapy had no effect on anxiety, except in patients treated with IL-2 in combination with INFalpha-2b. In these patients, the enhancement in anxiety scores that was observed on day 5 was mainly attributable to increased somatic complaints. CONCLUSION: IL-2 and INFalpha-2b have differential effects on mood, and IL-2 therapy induces depressive symptoms early in treatment. PMID- 10811681 TI - Primary cutaneous B-cell lymphoma: review and current concepts. AB - PURPOSE: Primary cutaneous B-cell lymphoma (PCBCL) has only recently been recognized as a distinct clinical entity. With the advent of improved immunophenotyping and immunogenotyping, increasing numbers of PCBCL cases are being diagnosed. However, there is much confusion regarding the classification, treatment, and prognosis of these patients. The purpose of this article is to review and analyze the available data to provide the clinician with a concise summary of the diagnosis, prognosis, and treatment of PCBCL. DESIGN: We conducted a thorough review of the medical literature on PCBCL, with a focus on classification, prognosis, and treatment trials. RESULTS AND CONCLUSION: PCBCL is defined as a B-cell lymphoma originating in the skin. There is no evidence of extracutaneous disease at presentation and for 6 months after diagnosis, as assessed by adequate staging procedures. Currently, the European Organization for Research and Treatment of Cancer classification is the most concise disease classification scheme, dividing the subtypes of PCBCL by clinical behavior and histopathologic findings. Based on this classification, the most common subtype of PCBCL is follicular center cell lymphoma. PCBCL is generally an indolent form of lymphoma with a good prognosis. Although local cutaneous recurrences are observed in 25% to 68% of patients, dissemination to internal organs is rare. Five-year survival rates typically range from 89% to 96%. A specific subtype, large B-cell lymphoma of the leg, is noted to have a poorer prognosis, with a 5 year survival rate of 58%. Overly aggressive treatment of PCBCL has not been shown to improve survival or prevent relapse. The treatment of choice usually varies depending on the type of PCBCL, the body surface area, and the location of the involvement, as well as the age and general health condition of the patient. The majority of studies indicate that PCBCL is highly responsive to radiation therapy. Polychemotherapy should be reserved for involvement of noncontiguous anatomic sites or those with extracutaneous spread. PMID- 10811682 TI - Thromboembolic events during chemotherapy for germ cell cancer: a cohort study and review of the literature. AB - PURPOSE: To evaluate the risk of major thromboembolic complications in male germ cell cancer patients receiving cisplatin-based chemotherapy and to review the literature on this subject. PATIENTS AND METHODS: One hundred seventy-nine germ cell cancer patients treated between January 1979 and May 1997 in our hospital were analyzed with respect to risk factors for developing thromboembolic events, such as baseline tumor characteristics, prior tumor therapy, administration of cytostatic agents, and the use of antiemetic drugs. The patients were treated with a variety of combination chemotherapy regimens, primarily cisplatin containing combination regimens. RESULTS: Of the 179 patients, 15 patients (8.4%) were identified who developed a total of 18 major thromboembolic complications in the time period between the start of chemotherapy and 6 weeks after administration of the last cytostatic drug in first-line treatment. Of these 18 events, three (16.7%) were arterial events, including two cerebral ischemic strokes, and 15 (83. 3%) were venous thromboembolic events, including 11 pulmonary embolisms. One (5.6%) of the 18 events was fatal. Liver metastases (odds ratio, 4.9; 95% confidence interval, 1.1 to 20.8) and the administration of high doses of corticosteroids (>/= 80 mg dexamethasone per cycle; odds ratio, 3.5; 95% confidence interval, 1. 2 to 10.3) as antiemetic therapy were identified as risk factors for the development of major thromboembolic complications. CONCLUSION: Germ cell cancer patients who receive chemotherapy, in particular those who have liver metastases or receive high doses of corticosteroids, are at considerable risk of developing thromboembolic complications. PMID- 10811683 TI - Evaluation of tumor measurements in oncology: use of film-based and electronic techniques. AB - PURPOSE: To evaluate the variability in bidimensional computed tomography (CT) measurements obtained of actual tumors and of tumor phantoms by use of three measurement techniques: hand-held calipers on film, electronic calipers on a workstation, and an autocontour technique on a workstation. MATERIALS AND METHODS: Three radiologists measured 45 actual tumors (in the lung, liver, and lymph nodes) on CT images, using each of the three techniques. Bidimensional measurements were recorded, and their cross-products calculated. The coefficient of variation was calculated to assess interobserver variability. CT images of 48 phantoms were measured by three radiologists with each of the techniques. In addition to the coefficient of variation, the differences between the cross product measurements of tumor phantoms themselves and the measurements obtained with each of the techniques were calculated. RESULTS: The differences between the coefficients of variation were statistically significantly different for the autocontour technique, compared with the other techniques, both for actual tumors and for tumor phantoms. There was no statistically significant difference in the coefficient of variation between measurements obtained with hand-held calipers and electronic calipers. The cross-products for tumor phantoms were 12% less than the actual cross-product when calipers on film were used, 11% less using electronic calipers, and 1% greater using the autocontour technique. CONCLUSION: Tumor size is obtained more accurately and consistently between readers using an automated autocontour technique than between those using hand-held or electronic calipers. This finding has substantial implications for monitoring tumor therapy in an individual patient, as well as for evaluating the effectiveness of new therapies under development. PMID- 10811684 TI - Temozolomide for advanced, metastatic melanoma. PMID- 10811685 TI - Second malignancies in Hodgkin's disease. PMID- 10811686 TI - Neural differentiation and prognosis in peripheral primitive neuroectodermal tumor. PMID- 10811687 TI - Chemotherapy of gestational trophoblastic disease. PMID- 10811688 TI - Dopamine agonists: their role in the treatment of Parkinson's disease. PMID- 10811689 TI - Botulinum toxin in muscle spasticity. PMID- 10811690 TI - Impaired cognitive performance in drug free users of recreational ecstasy (MDMA) PMID- 10811691 TI - Neurovisual rehabilitation: recent developments and future directions. PMID- 10811692 TI - Botulinum toxin (Dysport) treatment of hip adductor spasticity in multiple sclerosis: a prospective, randomised, double blind, placebo controlled, dose ranging study. AB - OBJECTIVE: To define a safe and effective dose of Dysport for treating hip adductor spasticity. METHODS: Patients with definite or probable multiple sclerosis, and disabling spasticity affecting the hip adductor muscles of both legs, were randomised to one of four treatment groups. Dysport (500, 1000, or 1500 Units), or placebo was administered by intramuscular injection to these muscles. Patients were assessed at entry, and 2, 4 (primary analysis time-point), 8, and 12 weeks post-treatment. RESULTS: A total of 74 patients were recruited. Treatment groups were generally well matched at entry. The primary efficacy variables-passive hip abduction and distance between the knees-improved for all groups. The improvement in distance between the knees for the 1500 Unit group was significantly greater than placebo (p = 0.02). Spasm frequency was reduced in all groups, but muscle tone was reduced in the Dysport groups only. Pain was reduced in all groups, but improvements in hygiene scores were evident only in the 1000 Unit and 1500 Unit groups. Duration of benefit was significantly longer than placebo for all Dysport groups (p<0.05). Adverse events were reported by 32/58 (55%) Dysport patients, and by 10/16 (63%) placebo patients. Compared with the two lower dose groups, twice as many adverse events were reported by the 1500 Unit group (2.7/patient). The incidence of muscle weakness was higher for the 1500 Unit group (36%) than for placebo (6%). The response to treatment was considered positive by two thirds of the patients in the 500 Unit group, and by about half the patients in the other groups. CONCLUSION: Dysport reduced the degree of hip adductor spasticity associated with multiple sclerosis, and this benefit was evident despite the concomitant use of oral antispasticity medication and analgesics. Although evidence for a dose response effect was not statistically significant, there was a clear trend towards greater efficacy and duration of effect with higher doses of Dysport. Dysport treatment was well tolerated, with no major side effects seen at doses up to 1500 Units. The optimal dose for hip adductor spasticity seems to be 500-1000 Units, divided between both legs. PMID- 10811693 TI - One year follow up study of primary and transitional progressive multiple sclerosis. AB - OBJECTIVE: To document clinical and magnetic resonance imaging (MRI) characteristics of a large cohort of primary and transitional progressive multiple sclerosis (PP and TP MS) patients over one year. INTRODUCTION: Patients with PP or TP MS have been shown to have low brain T2 and T1 lesion loads and slow rates of new lesion formation with minimal gadolinium enhancement, despite their accumulating disability. Serial evaluation of these patients is needed to elucidate the pathological processes responsible for disease progression and to identify clinical and MRI measures which can monitor these processes in treatment trials. METHOD: Patients, recruited from six European centres, underwent two assessments on the expanded disability status scale (EDSS) and MRI of the brain and spinal cord, 1 year apart. RESULTS: Of the 167 patients studied (137 with PP MS and 30 with TP MS), 41 (25%; 35 PP and six TP) showed a one step increase in the EDSS. The mean number of new brain lesions seen was 0.88 in the PP group and 0.47 in the TP MS group. Both groups demonstrated change in T2 lesion load over the year (p< or =0.002), with median percentage changes of 7.3% in the PP group and 10. 8% in the TP MS group. The PP group also showed a significant change in T1 load (p< 0.001, median change 12.6%). The number of new cord lesions seen was small (mean of 0.14 in the PP group and no new cord lesions in the TP group). Both groups demonstrated a decrease in cord cross sectional area (p< 0.001, median changes; PP 3.8%, TP 4. 9%), but only the PP group showed evidence of significant brain atrophy (p<0.001, 0.95%). CONCLUSION: Although the monitoring of disease progression in this patient group is difficult, this study demonstrates changes in both lesion load and atrophy, which, if shown to correlate with clinical change over a longer time will facilitate therapeutic trial design. PMID- 10811694 TI - Impaired cognitive performance in drug free users of recreational ecstasy (MDMA) AB - OBJECTIVES: Ecstasy (3,4-methylenedioxymethamphetamine (MDMA) and related congerers: MDA, MDEA) is the name given to a group of popular recreational drugs. Animal data raise concern about neurotoxic effects of high doses of ecstasy on central serotonergic systems. The threshold dose for neurotoxicity in humans is not clear and serotonin is involved in several functions including cognition. The purpose of this study was to investigate cognitive performance in a group of typical recreational ecstasy users. METHODS: A comprehensive cognitive test battery was administered to 28 abstinent ecstasy users with concomitant use of cannabis only and to two equally sized matched groups of cannabis users and non users. The sample consisted of ecstasy users with a typical recreational use pattern and did not include very heavy users. RESULTS: Ecstasy users were unimpaired in simple tests of attention (alertness). However, they performed worse than one or both control groups in the more complex tests of attention, in memory and learning tasks, and in tasks reflecting aspects of general intelligence. Heavier ecstasy and heavier cannabis use were associated with poorer performance in the group of ecstasy users. By contrast, the cannabis users did not differ significantly in their performance from the non-users. CONCLUSIONS: The present data raise concern that use of ecstasy possibly in conjunction with cannabis may lead to cognitive decline in otherwise healthy young people. Although the nature of the emerging cognitive disturbance is not yet clear, an impairment of working memory might be the common denominator underlying or contributing to declines of performance in various tasks. The cognitive disturbance is likely to be related to the well recognised neurotoxic potential of ecstasy. The data suggest that even typical recreational doses of ecstasy are sufficient to cause neurotoxicity in humans. PMID- 10811695 TI - Frontotemporal dementia and neuropsychology: the value of missing values. AB - OBJECTIVE: To investigate the effect of missing values due to behavioural disturbances on the neuropsychological test profile in frontotemporal dementia (FTD). The neuropsychological examination of patients with FTD poses a methodological problem. In many patients it is impossible to administer a complete test battery, due to behavioural disturbances inherent to the disease. This study describes the test behaviour of patients with FTD, the number of missing values due to disturbed test behaviour, and its influence on neuropsychological test results. METHODS: Thirty one patients with probable FTD were administered a neuropsychological test battery including measures of memory, intelligence, and executive functioning. Moreover, patients were rated on a global deterioration scale and a test behaviour scale, constructed for this study. RESULTS: The more disturbing the test behaviour, the less tests were assessable, leading to many missing values. The most disturbing features were "positive symptoms" of FTD, such as perseveration and stimulus boundedness. The effect of test behaviour was largest for tests measuring executive functions and reasoning capabilities. The replacement of the missing values due to behavioural disturbances by the lowest score also showed the largest effect on tests of executive function and reasoning abilities. CONCLUSION: Data imputation of missing values due to test behaviour disturbances provides a more differentiated picture of cognitive deficits in FTD. PMID- 10811696 TI - Group and case study of the dysexecutive syndrome in alcoholism without amnesia. AB - OBJECTIVES: To test the dysexecutive syndrome (DES) hypothesis of chronic alcoholism by the neuropsychological group and case study approaches. METHODS: A comprehensive neuropsychological assessment, including the "behavioural assessment of dysexecutive syndrome", a battery of tests recently designed to be "ecologically valid", was administered to 17 patients with chronic alcoholism without amnesia to examine executive functions, intelligence, and memory. In terms of each neuropsychological measure, reciprocal analyses of group means and individual case profiles were conducted: for the first contrasting the alcoholic patients with 17 age matched healthy subjects; and for the second making intersubject and intrasubject comparison of the patients, according to percentile basis impairment indices obtained from the control subjects. RESULTS: Despite relatively unimpaired memory and intelligence, the patients as a whole had the impairment of a wide range of executive domains, extending to "everyday" problem solving as well as more elementary aspects of executive functions, such as visuospatial performance, mental set shifting, and the inhibition of habitual behaviour. The profile analysis divided individual patients into four groups: the representative DES characterised by a clear dissociation between impaired executive functions and preserved intelligence and memory; the group of a modified dysexecutive pattern in which memory as well as executive functions were impaired with intelligence preserved; the group of general cognitive deterioration; and the group of unimpaired cognitive functioning. About two thirds of the patients were categorised into either the first or the second type of DES. CONCLUSION: DES characterised by the even more pronounced impairment of executive functions than of intelligence and memory afflicts a considerable proportion of patients with chronic alcoholism. Due to its subtlety, this would be potentially left out, unless appropriate behavioural measures were administered. This condition may prevent patients with alcoholism from achieving full recovery and benefiting from rehabilitation. PMID- 10811697 TI - Adult onset tic disorders. AB - BACKGROUND: Tic disorders presenting during adulthood have infrequently been described in the medical literature. Most reports depict adult onset secondary tic disorders caused by trauma, encephalitis, and other acquired conditions. Only rare reports describe idiopathic adult onset tic disorders, and most of these cases represent recurrent childhood tic disorders. OBJECTIVE: To describe a large series of patients with tic disorders presenting during adulthood, to compare clinical characteristics between groups of patients, and to call attention to this potentially disabling and underrecognised neurological disorder. METHODS: Using a computerised database, all patients with tic disorders who presented between 1988 and 1998 to the movement disorders clinic at Columbia-Presbyterian Medical Center after the age of 21 were identified. Patients' charts were retrospectively reviewed for demographic information, age of onset of tics, tic phenomenology, distribution, the presence of premonitory sensory symptoms and tic suppressibility, family history, and associated psychiatric features. These patients' videotapes were reviewed for diagnostic confirmation and information was obtained about disability, course, and response to treatment in a structured follow up interview. RESULTS: Of 411 patients with tic disorders in the database, 22 patients presented for the first time with tic disorders after the age of 21. In nine patients, detailed questioning disclosed a history of previous childhood transient tic disorder, but in 13 patients, the adult onset tic disorder was new. Among the new onset cases, six patients developed tics in relation to an external trigger, and could be considered to have secondary tic disorders. The remaining patients had idiopathic tic disorders. Comparing adult patients with recurrent childhood tics and those with new onset adult tics, the appearance of the tic disorder, the course and prognosis, the family history of tic disorder, and the prevalence of obsessive-compulsive disorder were found to be similar. Adults with new onset tics were more likely to have a symptomatic or secondary tic disorder, which in this series was caused by infection, trauma, cocaine use, and neuroleptic exposure. CONCLUSIONS: Adult onset tic disorders represent an underrecognised condition that is more common than generally appreciated or reported. The clinical characteristics of adults newly presenting to a movement disorder clinic with tic disorders are reviewed, analysed, and discussed in detail. Clinical evidence supports the concept that tic disorders in adults are part of a range that includes childhood onset tic disorders and Tourette's syndrome. PMID- 10811698 TI - Age specific prevalence of impairment and disability relating to hemiplegic stroke in the Hai District of northern Tanzania. Adult Morbidity and Mortality Project. AB - OBJECTIVES: To determine the age specific prevalence of impairment and disability relating to hemiplegic stroke in one rural area of Tanzania. METHODS: During the yearly house to house census of the study population of 148 135 (85 152 aged 15 and over) in August 1994, specific questions were asked to identify those who might be disabled from stroke. People thus identified were subsequently interviewed and examined by one investigator. In those in whom the clinical diagnosis of stroke was confirmed a more detailed interview and examination relating to risk factors and recovery was carried out. RESULTS: One hundred and eight patients, 61 men and 47 women, were identified with a median age of 70 (range 18-100). Median age at first stroke was 65 years. The age specific rates in this study were lower than previous studies in developed countries. All were cared for at home although 23 (21%) were bedbound. CONCLUSIONS: Although prevalence of impairment and disability related to stroke in this population as a whole was low this is mainly explained by the age structure, with less than 6% being aged 65 and over. Age standardised rates for stroke with residual disability were about half those found in previous studies in developed countries. Death from stroke in Africa may be higher but data are limited. With the demographic transition stroke is likely to become a more important cause of disability in sub-Saharan Africa. PMID- 10811699 TI - Neuromuscular findings in thyroid dysfunction: a prospective clinical and electrodiagnostic study. AB - OBJECTIVES: To evaluate neuromuscular signs and symptoms in patients with newly diagnosed hypothyroidism and hyperthyroidism. METHODS: A prospective cohort study was performed in adult patients with newly diagnosed thyroid dysfunction. Patients were evaluated clinically with hand held dynamometry and with electrodiagnosis. The clinical features of weakness and sensory signs and the biochemical data were evaluated during treatment. RESULTS: In hypothyroid patients 79% had neuromuscular complaints, 38% had clinical weakness (manual muscle strength testing) in one or more muscle groups, 42% had signs of sensorimotor axonal neuropathy, and 29% had carpal tunnel syndrome. Serum creatine kinase did not correlate with weakness. After 1 year of treatment 13% of the patients still had weakness. In hyperthyroid patients 67% had neuromuscular symptoms, 62% had clinical weakness in at least one muscle group that correlated with FT4 concentrations, but not with serum CK. Nineteen per cent of the patients had sensory-motor axonal neuropathy and 0% had carpal tunnel syndrome. The neuromuscular signs developed rapidly, early in the course of the disorder and were severe, but resolved rapidly and completely during treatment (average time 3.6 months). CONCLUSIONS: Neuromuscular symptoms and signs were present in most patients. About 40% of the hypothyroid patients and 20% of the hyperthyroid patients had predominantly sensory signs of a sensorimotor axonal neuropathy early in the course of thyroid disease. Weakness in hyperthyroidism evolved rapidly at an early stage of the disorder and resolved completely during treatment, suggesting a functional muscle disorder. Hand held dynamometry is sensitive for the detection of weakness and for the clinical evaluation of treatment effects. Weakness in hypothyroidism is more difficult to treat, suggesting myopathy. PMID- 10811700 TI - Evaluation of three different ways of assessing tremor in multiple sclerosis. AB - OBJECTIVES: To examine the comparative reliability and validity of three simple ways of rating upper limb tremor in patients with multiple sclerosis. METHODS: Three examiners independently rated severity of upper limb tremor in patients with multiple sclerosis on a 0-10 scale by studying videotape recordings of patients' examinations, spiral drawings, and handwriting samples. The correlations of the tremor severity scores with scores from arm dexterity tests and a tremor related disability scale were also assessed. RESULTS: Rating tremor on posture had a good intrarater and interrater reliability. However, these reliabilities decreased when kinetic tremor was assessed, in part because dysmetria was a confounding factor. The intrarater reliabilities of rating tremor from spirals and handwriting were also good but the interrater reliabilities were only fair to moderate. Tremor severity scored by all three methods correlated highly with scores obtained from the nine hole peg test, finger tapping test, and a tremor related activities of daily living (ADL) questionnaire, indicating that all three methods were valid ways of assessing tremor in multiple sclerosis. CONCLUSION: Multiple sclerosis tremors in posture can be scored using a clinical rating scale in a valid and reliable way, and from spirals and handwriting samples if the ratings are carried out by the same examiner. However, scoring kinetic tremor was less reliable. In addition, the nine hole peg and finger tapping tests provide useful objective assessments of upper limb function in tremulous patients with multiple sclerosis. PMID- 10811702 TI - Invasive aspergillosis in a patient with MELAS syndrome. AB - Invasive infection with the opportunistic fungus Aspergillus fumigatus predominantly affects people with impaired cell mediated immunity. The case of a 31 year old woman with no identified cause for immunosuppression who presented with severe refractory aspergillosis of the paranasal sinuses is reported. She subsequently developed clinical and molecular evidence of mitochondrial encephalomyopathy with lactic acidosis and stroke-like events (MELAS) syndrome. It is proposed that MELAS syndrome may represent an unusual risk factor for the development of invasive aspergillosis and mechanisms are supported by which mitochondrial dysfunction may predispose to this. PMID- 10811701 TI - Reversible neuropsychological deficits after mild traumatic brain injury. AB - OBJECTIVES: To determine the influence of motivation on performance in a divided attention test of patients after mild traumatic brain injury (MBI). METHODS: Comparison of the performance of 12 patients with MBI with 10 patients with severe brain injury (SBI) and 11 healthy controls in a computer supported divided attention task before (T1) and after (T2) verbal motivation. RESULTS: At T1, the MBI group performed the same as the SBI group but significantly worse than the controls in all variables. At T2, the MBI group performed worse than the controls at T2 but the results were equal to the results of the controls at T1 and significantly better than the SBI group at T1 or T2. At T2 the MBI group performed at the level of published norms for the rest. CONCLUSION: Before verbal motivation the MBI group's results in the divided attention task were comparable with those from patients with severe brain injury. They failed to exploit their performance potential when it depended on self motivation but were able to perform at the level of the control group when external motivation was applied. PMID- 10811703 TI - Manometric investigation of anorectal function in early and late stage Parkinson's disease. AB - Abnormal gastrointestinal function is relatively frequent in Parkinson's disease, and constipation is a disturbing symptom in many patients. However, it remains to be established whether anorectal abnormalities are characteristic of the late stages of the disease. Clinical and anorectal manometric function were investigated in groups of early and late stage parkinsonian patients. Thirty one patients (19 men, 12 women, age range 22 to 89 years) entered the study. The disease severity was assessed by Hoehn and Yahr staging: there were four (12.9%) stage I, seven (22.6%) stage II, 10 (32.2%) stage III, and 10 (32.2%) stage IV patients. Anorectal variables were measured by standard manometric equipment and techniques. Values obtained in early stage patients (Hoehn and Yahr stage I and II) were compared with those obtained in late stage patients (Hoehn and Yahr stage III and IV). Overall, more than 70% of patients complained of chronic constipation, with chronic laxative use reported in more than 30%. Late stage patients were slightly older than their early stage counterparts. Pelvic floor dyssynergia was documented in more than 60% of patients. Manometric variables were not different in the two groups. In conclusion, defecatory dysfunction is frequent in Parkinson's disease, it is not confined to late stage patients, and it is found early in the course of the disease. This has potential implications for a targeted therapeutic approach. PMID- 10811704 TI - Postpartum lumbosacral plexopathy limited to autonomic and perineal manifestations: clinical and electrophysiological study of 19 patients. AB - The objective was to describe perineal electrophysiological findings and to determine their diagnostic value in a type of lumbosacral plexopathy after vaginal delivery, which only involves the lower part of the plexus (S2-S4). Consecutive female patients referred to an outpatients' urodynamic clinic were the source. Nineteen previously healthy women, 13 multiparae and six para 1, were investigated. Mean age was 33.7 (SD 5.4) (range 28-41) years. All of them presented with urinary (stress incontinence 14, dysuria five), anorectal (faecal incontinence eight, dyskesia one), or sexual dysfunctions (hypoorgasmia or anorgasmia six) after vaginal delivery. No associated lower limb sensory or motor deficits were noted. All the patients had electrophysiological recordings (bulbocavernosus muscle EMG, measurements of the bulbocavernosus reflex latencies (BCRLs), somatosensory evoked potentials of the pudendal nerve (SEPPNs), and pudendal nerve terminal motor latencies (PNTMLs)). Cystometry and urethral pressure profile (UPP) were performed in the 14 patients with stress urinary incontinence. Perineal electrophysiological examination disclosed signs of denervation in the perineal muscles in all the cases, prolonged BCRLs in 17/19, and abolished BCRLs in 2/19, abnormal SEPPN in 1/19, and normal PNTMLs in all the patients. Urodynamic investigations disclosed low urethral closure pressure for age (< 50 cm H(2)O) in half of the patients. In conclusion, Lower postpartum lumbosacral plexopathy is evoked when perineal sensory disturbances whether or not associated with urinary or faecal incontinence persist after a history of a difficult vaginal delivery. Electrophysiological investigations precisely identify the site of the lesion and demonstrate distal innervation integrity. PMID- 10811705 TI - Fatal familial insomnia: clinical, neuropathological, and genetic description of a Spanish family. AB - The clinical presentation and evolution, neuropathological findings, and genotyping of three members of a Spanish family affected with fatal familial insomnia are reported. The mother and two of her offspring developed a rapidly evolving disease with insomnia and behavioural disorders as the initial symptoms and died between 5 and 10 months after the onset of the illness. Frontal brain biopsy in the mother disclosed only non-significant spongiosis, and full neuropathological examination of her offspring showed thalamic and olivary degeneration with isolated focal cortical spongiosis. Genetic examination could only be performed in the contemporary patients and both harboured the prion protein (PrP) 178Asn mutation and homozygous 129 Met/Met genotype. PMID- 10811707 TI - August forel (1848-1931) PMID- 10811706 TI - Alzheimer's disease comorbidity in normal pressure hydrocephalus: prevalence and shunt response. AB - The clinical impact of Alzheimer's disease pathology at biopsy was investigated in 56 cognitively impaired patients undergoing shunt surgery for idiopathic normal pressure hydrocephalus (NPH). Cognition was measured by means of the global deterioration scale (GDS), the mini mental status examination (MMSE) and a battery of six psychometric tests. Gait was assessed using objective measurements of velocity and the ambulatory index (AI). The prevalence of cases exhibiting neuritic plaques (positive biopsies) increased in parallel with dementia severity from 18% for patients with GDS 3 to 75% for patients with GDS scores > or =6. Patients with positive biopsies were more cognitively impaired (higher GDS and lower MMSE scores) as well as more gait impaired (higher AI scores and slower velocities) than patients with negative biopsies. After surgery, gait velocity and AI scores improved significantly and to a comparable degree for patients with and without positive biopsies. Similar proportions of positive and negative biopsy patients also had improved gait as assessed by means of subjective video tape comparisons. There were no significant differences between the biopsy groups in the magnitude of postoperative psychometric change or in the proportion of cases exhibiting improved urinary control. Alzheimer's disease pathology is a common source of comorbidity in older patients with idiopathic NPH where it contributes to the clinical impairment associated with this disorder. For patients accurately diagnosed with NPH, concomitant Alzheimer's disease pathology does not strongly influence the clinical response to shunt surgery. PMID- 10811708 TI - Intact verbal description of letters with diminished awareness of their forms. AB - Visual processing and its conscious awareness can be dissociated. To examine the extent of dissociation between ability to read characters or words and to be consciously aware of their forms, reading ability and conscious awareness for characters were examined using a tachistoscope in an alexic patient. A right handed woman with 14 years of education presented with incomplete right hemianopia, alexia with kanji (ideogram) agraphia, anomia, and amnesia. Brain MRI disclosed cerebral infarction limited to the left lower bank of the calcarine fissure, lingual and parahippocampal gyri, and an old infarction in the right medial frontal lobe. Tachistoscopic examination disclosed that she could read characters aloud in the right lower hemifield when she was not clearly aware of their forms and only noted their presence vaguely. Although her performance in reading kanji was better in the left than the right field, she could read kana (phonogram) characters and Arabic numerals equally well in both fields. By contrast, she claimed that she saw only a flash of light in 61% of trials and noticed vague forms of stimuli in 36% of trials. She never recognised a form of a letter in the right lower field precisely. She performed judgment tasks better in the left than right lower hemifield where she had to judge whether two kana characters were the same or different. Although dissociation between performance of visual recognition tasks and conscious awareness of the visual experience was found in patients with blindsight or residual vision, reading (verbal identification) of characters without clear awareness of their forms has not been reported in clinical cases. Diminished awareness of forms in our patient may reflect incomplete input to the extrastriate cortex. PMID- 10811709 TI - Trends in mortality from sporadic Creutzfeldt-Jakob disease in France 1992-7. AB - This study examined trends in mortality from sporadic Creutzfeldt-Jakob disease in France for 1992-7 by age, genotype at the codon 129 of the prion protein gene, and geographical area. Case ascertainment was based on notifications by neurologists, neuropathologists, and laboratories; 324 deaths from definite or probable Creutzfeldt-Jakob disease were registered during the study period. The yearly number of deaths increased significantly between 1992 and 1997. The rise was higher for older age groups. It was also higher in those who were homozygous for valine compared with other genotypes. Eighteen departments (geographical administrative areas) out of 95 showed a significant increase in the number of deaths from sporadic Creutzfeldt-Jakob disease. Intensive epidemiological surveillance is a likely explanation for the apparent increase in the number of sporadic cases of Creutzfeldt-Jakob disease over the 1992-7 period, particularly in older age groups and in departments with low mortality rate at the beginning of the study period. Intensive surveillance may also have led to a better identification of atypical valine homozygous cases. PMID- 10811710 TI - Cerebral infarction complicating intravenous immunoglobulin therapy in a patient with Miller Fisher syndrome. AB - Intravenous immunoglobulin (IVIg) therapy is being increasingly used in a wide range of neurological conditions. However, treatment is expensive and side effects may be severe. A patient with Miller Fisher syndrome who developed cortical blindness as a consequence of occipital infarction precipitated by IVIg is reported on. PMID- 10811711 TI - Recanalisation of spinal dural arteriovenous fistula after successful embolisation. PMID- 10811713 TI - Botulinum toxin in the treatment of cerebral palsy PMID- 10811712 TI - Is transcranial colour Duplex flow imaging of use in selection of patients with acute stroke for thrombolysis? PMID- 10811714 TI - McAlpine's multiple sclerosis. Third edition PMID- 10811715 TI - MR imaging of the brachial plexus PMID- 10811716 TI - A textbook of SPECT in neurology and psychiatry PMID- 10811717 TI - Challenge epilepsy-New antiepileptic drugs PMID- 10811718 TI - Advances in multiple sclerosis-clinical research and therapy PMID- 10811719 TI - Receptors, gephyrin and gephyrin-associated proteins: novel insights into the assembly of inhibitory postsynaptic membrane specializations. AB - The synaptic localization of ion channel receptors is essential for efficient synaptic trans-mission and the precise regulation of diverse neuronal functions, such as signal integration and synaptic plasticity. Emerging evidence points to an important role of cytoskeleton-associated proteins that assemble receptors and components of the subsynaptic machinery at postsynaptic membrane specializations. This article reviews interactions of inhibitory postsynaptic neurotransmitter receptors with the receptor anchoring protein gephyrin and intracellular components involved in downstream signalling and/or control of signal transduction processes. The presently available data suggest a central synaptic organizer function for gephyrin in inhibitory postsynaptic membrane assembly and stabilization. PMID- 10811720 TI - Regulation of neuronal function by protein trafficking: a role for the endosomal pathway. AB - Protein trafficking plays a central role in many aspects of neuronal function, from the release of neurotransmitters by exocytosis and the recycling of synaptic vesicle proteins to the regulation of receptor signalling. Synaptic function can be significantly modified on a short time scale by alterations in the levels of receptors, ion channels and transporters both pre- and postsynaptically. In many cases, these alterations appear to be mediated by acute changes in the rates at which the proteins are endocytosed from and exocytosed to the cell surface from intracellular pools. While our current understanding of the signalling mechanisms and the intracellular pathways responsible for these acute changes is still in its infancy, intriguing details are beginning to emerge from a number of systems. PMID- 10811721 TI - Receptor targeting and heterogeneity at interneuronal nicotinic cholinergic synapses in vivo. AB - Within a single neuron the correct targeting of the diverse neurotransmitter receptor types to discrete synaptic regions is crucial for proper function. However, the molecular mechanisms that underlie neuronal receptor clustering and targeting are still largely undefined. Here we report advances in defining the mechanisms that mediate nicotinic acetylcholine receptor (nAChR) targeting to interneuronal synapses. Recent in vivo studies have demonstrated that one subunit plays a critical role in the differentiation of nicotinic cholinergic synapses on vertebrate autonomic neurons. The major cytoplasmic loop of the alpha3 subunit targets specific nAChR subtypes to the synapse. In contrast, nAChR complexes that lack the alpha3 targeting domain are excluded and are perisynaptic. Additional studies have demonstrated a greater complexity to alpha3-nAChR targeting due to a unique postsynaptic receptor microheterogeneity - under one presynaptic terminal, alpha3-nAChR clusters are separate, but proximal to, glycine receptor (GlyR) clusters in discrete postsynaptic membrane microregions. The surprising coexistence under one nerve ending of separate clusters of receptors that respond to different fast-acting transmitters with opposing functions may represent a novel mechanism for modulating synaptic activity. Overall, the receptor targeting and clustering studies reviewed in this issue suggest that a common mechanism underlies the formation of the diverse types of interneuronal synapses but differs from that responsible for neuromuscular junction assembly in vertebrates. PMID- 10811722 TI - Synaptic efficacy and reliability of excitatory connections between the principal neurones of the input (layer 4) and output layer (layer 5) of the neocortex. AB - A prerequisite for the understanding of how a cortical column functions is a description of small and defined neuronal circuits consisting of only a few identified neurones. Here we summarise, with particular reference to the barrel cortex, the morphological and physiological properties of two synaptic connections, namely those between pairs of spiny neurones in layer 4 and pairs of pyramidal cells in layer 5. While layer 4 spiny neurones are the cortical input neurones that amplify and relay incoming excitation from the periphery, layer 5 pyramidal cells integrate neuronal activity both within and across cortical columns and subsequently distribute it to both cortical and subcortical brain regions. PMID- 10811723 TI - Target-specific expression of pre- and postsynaptic mechanisms. AB - Target-specific expression of pre- and postsynaptic mechanisms of synaptic transmission has been shown in a variety of central neurons by a number of laboratories. These data have demonstrated that synaptic transmission between single axons diverging onto distinct target neurons can behave independently, differentially influencing activity in the target neuron. Similarly, single neurons are capable of manufacturing molecularly distinct ligand-gated receptors and targeting them to synapses innervated by distinct converging afferent projections. A picture is emerging consistent with a role for both pre- and postsynaptic mechanisms in influencing the target-specific nature of transmission at numerous diverse synapses throughout the mammalian CNS. This target specificity adds another level of complexity in unravelling the roles played by individual neurons within a computational network. To begin to understand the coordinated activity of large ensembles of neurons it is becoming clear that the nature of transmission between individual pre- and postsynaptic elements within a circuit must first be understood for each and every neural element involved. PMID- 10811724 TI - Microheterogeneity of calcium signalling in dendrites. AB - Transient changes in the intracellular concentration of free Ca2+ ([Ca2+]i) originating from voltage- or ligand-gated influx and by ligand- or Ca2+-gated release from intracellular stores, trigger or modulate many fundamental neuronal processes, including neurotransmitter release and synaptic plasticity. Of the intracellular compartments involved in Ca2+ clearance, the endoplasmic reticulum (ER) has received the most attention because it expresses Ca2+ pumps and Ca2+ channels, thus endowing it with the potential to act as both an intracellular calcium sink and store. We review here our ongoing work on the role of calcium sequestration into, and release from, ER cisterns and the role that this plays in the generation and termination of free [Ca2+]i transients in dendrites of pyramidal neurons in hippocampal slices during and after synaptic activity. These studies have been approached by combining parallel microfluorometric measurements of free cytosolic [Ca2+]i transients with energy-dispersive X-ray microanalytical measurements of total Ca content within specific dendritic compartments at the electron microscopy level. Our observations support the emerging realization that specific subsets of dendritic ER cisterns provide spatial and temporal microheterogeneity of Ca2+ signalling, acting not only as a major intracellular Ca sink involved in active clearance mechanisms after voltage- and ligand-gated Ca2+ influx, but also as an intracellular Ca2+ source that can be mobilized by a signal cascade originating at activated synapses. PMID- 10811726 TI - Dendritic potassium channels in hippocampal pyramidal neurons. AB - Potassium channels located in the dendrites of hippocampal CA1 pyramidal neurons control the shape and amplitude of back-propagating action potentials, the amplitude of excitatory postsynaptic potentials and dendritic excitability. Non uniform gradients in the distribution of potassium channels in the dendrites make the dendritic electrical properties markedly different from those found in the soma. For example, the influence of a fast, calcium-dependent potassium current on action potential repolarization is progressively reduced in the first 150 micrometer of the apical dendrites, so that action potentials recorded farther than 200 micrometer from the soma have no fast after-hyperpolarization and are wider than those in the soma. The peak amplitude of back-propagating action potentials is also progressively reduced in the dendrites because of the increasing density of a transient potassium channel with distance from the soma. The activation of this channel can be reduced by the activity of a number of protein kinases as well as by prior depolarization. The depolarization from excitatory postsynaptic potentials (EPSPs) can inactivate these A-type K+ channels and thus lead to an increase in the amplitude of dendritic action potentials, provided the EPSP and the action potentials occur within the appropriate time window. This time window could be in the order of 15 ms and may play a role in long-term potentiation induced by pairing EPSPs and back propagating action potentials. PMID- 10811725 TI - Ion channel sequestration in central nervous system axons. AB - Na+ and K+ channel localization and clustering are essential for proper electrical signal generation and transmission in CNS myelinated nerve fibres. In particular, Na+ channels are clustered at high density at nodes of Ranvier, and Shaker-type K+ channels are sequestered in juxtaparanodal zones, just beyond the paranodal axoglial junctions. The mechanisms of channel localization at nodes of Ranvier in the CNS during development in both normal and hypomyelinating mutant animals are discussed and reviewed. As myelination proceeds, Na+ channels are initially found in broad zones within gaps between neighbouring oligodendroglial processes, and then are condensed into focal clusters. This process appears to depend on the formation of axoglial junctions. K+ channels are first detected in juxtaparanodal zones, and in mutant mice lacking normal axoglial junctions, these channels fail to cluster. In these mice, despite the presence of numerous oligodendrocytes, Na+ channel clusters are rare, and when present, are highly irregular. A number of molecules have recently been described that are candidates for a role in the neuron-glial interactions driving ion channel clustering. This paper reviews the cellular and molecular events responsible for formation of the mature node of Ranvier in the CNS. PMID- 10811727 TI - Effects of pH on the uncoupled, coupled and pre-steady-state currents at the amino acid transporter KAAT1 expressed in Xenopus oocytes. AB - The effects of pH on the different kinds of currents occurring at the lepidopteran amino acid cotransporter KAAT1 have been examined using heterologous expression in Xenopus oocytes and voltage clamp. Acidic pH (5.5-4.5) caused a slight depression of the uncoupled current and a complete inhibition of the coupled and transient currents, in the presence of either Na+ or K+, the two ions physiologically relevant to the transport process. Conversely, at alkaline pH (9) no statistically significant effects could be observed on uncoupled, coupled and transient currents compared to the effects at pH 7.6. These effects of pH indicate that operation of the transporter is maximal in the physiologically alkaline native environment. The dose-response curves for the inhibition of coupled and transient currents were similar, with respective pKa values of 6. 29 +/- 0.05 and 6.40 +/- 0.03 and respective Hill coefficient values (nH) of 0.93 +/ 0.07 and 1.08 +/- 0.08, suggesting that the two effects can be explained by a single proton binding to the same site in the transporter. PMID- 10811728 TI - Expression of ryanodine receptor RyR3 produces Ca2+ sparks in dyspedic myotubes. AB - Discrete, localized elevations of myoplasmic [Ca2+], Ca2+ 'sparks', were readily detected using the fluorescent Ca2+ indicator fluo-3 and laser scanning confocal microscopy in 'dyspedic' 1B5 myotubes, i.e. myotubes which do not express ryanodine receptors (RyRs), transduced with virions containing cDNA for RyR type 3 that were saponin permeabilized to allow dye entry. Ca2+ sparks were never observed in non-transduced RyR null myotubes. The spatial locations of sparks observed in permeabilized myotubes roughly corresponded to regions of RyR protein expression in the same myotube as detected after subsequent fixation and antibody staining. Permeabilized RyR3-transduced myotubes exhibited similar punctate peripheral RyR3 protein immunohistochemical patterns as myotubes fixed before permeabilization indicating that permeabilization did not affect the structural organization of the triad. Ca2+ sparks, recorded in line scan mode, in permeabilized myotubes expressing RyR3 exhibited mean amplitudes (change in fluorescence/mean fluorescence, DeltaF/F: 1.20 +/- 0.04) and temporal rise times (10-90%; 6.31 +/- 0.12 ms) similar to those of sparks recorded in permeabilized frog skeletal muscle fibres (0.98 +/- 0.01; 6.11 +/- 0.07, respectively) using the same confocal system. Spatial extent and temporal duration of the Ca2+ sparks were approximately 40% larger in the RyR3-expressing myotube cultures than in frog fibres. Ca2+ sparks recorded in line scan mode often occurred repetitively at the same spatial location in RyR3-expressing myotubes. Such repetitive events were highly reproducible in amplitude and spatio-temporal properties, as previously observed for repetitive mode sparks in frog skeletal muscle. Ca2+ sparks recorded in xy mode were frequently compressed in the y (slower scan) direction compared to the x direction. This asymmetry was reproduced assuming spatially symmetric events having the time course of Ca2+ sparks recorded in line scan (xt) mode. These expression studies demonstrate that the presence of RyR3 is sufficient for the production of Ca2+ sparks in a skeletal muscle system lacking the expression of any other RyR isoform. PMID- 10811729 TI - Properties of gastric smooth muscles obtained from mice which lack inositol trisphosphate receptor. AB - Membrane potential recordings, made from the circular smooth muscle layer of the gastric antrum taken from mutant mice which lacked the inositol trisphosphate (InsP3) type 1 receptor, were compared with those obtained from the stomach of control (wild-type) mice. Immunostaining of gastric muscles indicated that the distribution and form of c-kit positive cells were similar in wild-type and mutant mice. Smooth muscles from wild-type mice generated slow waves that in turn initiated spike potentials, while those from mutant mice were either quiescent or generated irregular bursts of spike potentials. In the presence of nifedipine, slow waves with reduced amplitude were generated in wild-type mice, while all electrical activity was abolished in mutant mice. Acetylcholine depolarized and sodium nitroprusside hyperpolarized the membrane in muscles from both types of mice, being more effective in wild-type mice. Noradrenaline produced similar hyperpolarizations in both types of mice. Transmural nerve stimulation evoked inhibitory junction potentials (IJPs) in both wild-type and mutant mice. In wild type mice, the IJPs were reduced in amplitude by nitroarginine and converted to a cholinergic excitatory junction potential (EJP) by apamin. In mutant mice, the IJPs were unaffected by nitroarginine or atropine but were abolished by apamin. It is concluded that in antral smooth muscle, the expression of InsP3 type 1 receptors may be causally related to the generation of slow waves but not to the generation of action potentials. A lack of InsP3 receptors attenuates cholinergic excitatory and nitrergic inhibitory responses but does not alter the response to noradrenaline. PMID- 10811730 TI - Two Ca2+ entry pathways mediate InsP3-sensitive store refilling in guinea-pig colonic smooth muscle. AB - Sarcolemma Ca2+ influx, necessary for store refilling, was well maintained, over a wide range (-70 to + 40 mV) of membrane voltages, in guinea-pig single circular colonic smooth muscle cells, as indicated by the magnitude of InsP3-evoked Ca2+ transients. This apparent voltage independence of store refilling was achieved by the activity of sarcolemma Ca2+ channels some of which were voltage gated while others were not. At negative membrane potentials (e.g. -70 mV), Ca2+ influx through channels which lacked voltage gating provided for store refilling while at positive membrane potentials (e.g. +40 mV) voltage-gated Ca2+ channels were largely responsible. Sarcolemma voltage-gated Ca2+ currents were not activated following store depletion. Removal of external Ca2+ or the addition of the Ca2+ channel blocker nimodipine (1 microM) inhibited store refilling, as assessed by the magnitude of InsP3-evoked Ca2+ transients, with little or no change in bulk average cytoplasmic Ca2+ concentration. One hypothesis for these results is that the store may refill from a high subsarcolemma Ca2+ gradient. Influx via channels, some of which are voltage gated and others which lack voltage gating, may permit the establishment of a subsarcolemma Ca2+ gradient. Store access to the gradient allows InsP3-evoked Ca2+ signalling to be maintained over a wide voltage range in colonic smooth muscle. PMID- 10811731 TI - Functional state of the plasma membrane Ca2+ pump in Plasmodium falciparum infected human red blood cells. AB - The active Ca2+ transport properties of malaria-infected, intact red blood cells are unknown. We report here the first direct measurements of Ca2+ pump activity in human red cells infected with Plasmodium falciparum, at the mature, late trophozoite stage. Ca2+ pump activity was measured by the Co2+-exposure method adapted for use in low-K+ media, optimal for parasitised cells. This required a preliminary study in normal, uninfected red cells of the effects of cell volume, membrane potential and external Na+/K+ concentrations on Ca2+ pump performance. Pump-mediated Ca2+ extrusion in normal red cells was only slightly lower in low K+ media relative to high-K+ media despite the large differences in membrane potential predicted by the Lew-Bookchin red cell model. The effect was prevented by clotrimazole, an inhibitor of the Ca2+-sensitive K+ (KCa) channel, suggesting that it was due to minor cell dehydration. The Ca2+-saturated Ca2+ extrusion rate through the Ca2+ pump (Vmax) of parasitised red cells was marginally inhibited (2 27 %) relative to that of both uninfected red cells from the malaria-infected culture (cohorts), and uninfected red cells from the same donor kept under identical conditions (co-culture). Thus, Ca2+ pump function is largely conserved in parasitised cells up to the mature, late trophozoite stage. A high proportion of the ionophore-induced Ca2+ load in parasitised red cells is taken up by cytoplasmic Ca2+ buffers within the parasite. Following pump-mediated Ca2+ removal from the host, there remained a large residual Ca2+ pool within the parasite which slowly leaked to the host cell, from which it was pumped out. PMID- 10811732 TI - An oxygen-, acid- and anaesthetic-sensitive TASK-like background potassium channel in rat arterial chemoreceptor cells. AB - The biophysical and pharmacological properties of an oxygen-sensitive background K+ current in rat carotid body type-I cells were investigated and compared with those of recently cloned two pore domain K+ channels. Under symmetrical K+ conditions the oxygen-sensitive whole cell K+ current had a linear dependence on voltage indicating a lack of intrinsic voltage sensitivity. Single channel recordings identified a K+ channel, open at resting membrane potentials, that was inhibited by hypoxia. This channel had a single channel conductance of 14 pS, flickery kinetics and showed little voltage sensitivity except at extreme positive potentials. Oxygen-sensitive current was inhibited by 10 mM barium (57% inhibition), 200 microM zinc (53% inhibition), 200 microM bupivacaine (55% inhibition) and 1 mM quinidine (105 % inhibition). The general anaesthetic halothane (1.5%) increased the oxygen-sensitive K+ current (by 176%). Halothane (3 mM) also stimulated single channel activity in inside-out patches (by 240%). Chloroform had no effect on background K+ channel activity. Acidosis (pH 6.4) inhibited the oxygen-sensitive background K+ current (by 56%) and depolarised type-I cells. The pharmacological and biophysical properties of the background K+ channel are, therefore, analogous to those of the cloned channel TASK-1. Using in situ hybridisation TASK-1 mRNA was found to be expressed in type-I cells. We conclude that the oxygen- and acid-sensitive background K+ channel of carotid body type-I cells is likely to be an endogenous TASK-1-like channel. PMID- 10811733 TI - Co-release of ATP and ACh mediates hypoxic signalling at rat carotid body chemoreceptors. AB - Using functional co-cultures of rat carotid body (CB) O2 chemoreceptors and 'juxtaposed' petrosal neurones (JPNs), we tested whether ATP and ACh acted as co transmitters. Perforated-patch recordings from JPNs often revealed spontaneous and hypoxia-evoked (PO2 approximately 5 mmHg) excitatory postsynaptic responses. The P2X purinoceptor blocker, suramin (50 microM) or a nicotinic ACh receptor (nAChR) blocker (hexamethonium, 100 microM; mecamylamine, 1 microM) only partially inhibited these responses, but together, blocked almost all activity. Under voltage clamp (-60 mV), fast perfusion of 100 microM ATP over hypoxia responsive JPNs induced suramin-sensitive (IC50 = 73 microM), slowly desensitizing, inward currents (IATP) with time constant of activation tauon = 30.6 +/- 4. 8 ms (n = 7). IATP reversed at 0.33 +/- 3.7 mV (n = 4), and the dose response curve was fitted by the Hill equation (EC50 = 2.7 microM; Hill coefficient approximately 0.9). These purinoceptors contained immunoreactive P2X2 subunits, but their activation by alpha,beta-methylene ATP (alpha,beta-meATP; EC50 = 2.1 microM) suggests they are P2X2/P2X3 heteromultimers. Suramin and nAChR blockers inhibited the extracellular chemosensory discharge in the intact rat carotid body-sinus nerve preparation in vitro. Further, P2X2 immunoreactivity was widespread in rat petrosal ganglia in situ, and co-localized in neurones expressing the CB chemo-afferent marker, tyrosine hydroxylase (TH). P2X2 labelling in the CB co-localized with nerve-terminal markers, and was intimately associated with TH-positive type 1 cells. Thus ATP and ACh are co-transmitters during chemotransduction in the rat carotid body. PMID- 10811734 TI - Nutrient modulation of polarized and sustained submembrane Ca2+ microgradients in mouse pancreatic islet cells. AB - The intracellular calcium concentration ([Ca2+]i) near the plasma membrane was measured in mouse pancreatic islet cells using confocal spot detection methods. Whereas small cytosolic Ca2+ gradients were observed with 3 mM glucose, a steeper sustained gradient restricted to domains beneath the plasma membrane (space constant, 0.67 micrometer) appeared with 16.7 mM glucose. When the membrane potential was clamped with increasing K+ concentrations (5, 20 and 40 mM), no [Ca2+]i gradients were observed in any case. Increasing glucose concentration (0, 5 and 16.7 mM) in the presence of 100 microM diazoxide, a K+ channel opener, plus 40 mM K+ induced steeper [Ca2+]i gradients, confirming the role of membrane potential-independent effects of glucose. Prevention of Ca2+ store refilling with 30 microM cyclopiazonic acid (CPA) or blockade of uniporter-mediated Ca2+ influx into the mitochondria with 1 microM carbonyl cyanide m-chlorophenyl hydrazone (CCCP) or 1 microM Ru-360 significantly reduced the steepness of the 16.7 mM glucose-induced [Ca2+]i gradients. Measured values of [Ca2+]i reached 6.74 +/- 0.67 microM at a distance of 0.5 micrometer from the plasma membrane and decayed to 0.27 +/- 0.03 microM at a distance of 2 micrometer. Mathematically processed values at 0.25 and 0 micrometer gave a higher [Ca2+]i, reaching 8.18 +/- 0.86 and 10.05 +/- 0.98 microM, respectively. The results presented indicate that glucose metabolism generates [Ca2+]i microgradients, which reach values of around 10 microM, and whose regulation requires the involvement of both mitochondrial Ca2+ uptake and endoplasmic reticulum Ca2+ stores. PMID- 10811735 TI - Ca2+ dependence of loaded shortening in rat skinned cardiac myocytes and skeletal muscle fibres. AB - This study examined the effects of activator Ca2+ on loaded shortening and power output in skinned rat cardiac myocyte preparations, and fast- and slow-twitch skeletal muscle fibres at 12 degrees C. Shortening velocities were slowed at nearly all relative loads when Ca2+ activation levels were reduced to approximately 70% maximal isometric force (P4.5) in cardiac myocyte preparations, as well as in fast-twitch and slow-twitch skeletal muscle fibres. Peak absolute power outputs declined significantly as Ca2+ activation levels were progressively reduced from maximal to 30% P4.5 in all three striated muscle types, with the greatest change in fast-twitch fibres. In cardiac myocyte preparations, even peak relative power output progressively fell when Ca2+ activation levels were lowered to approximately 70, 50 and 30% P4.5. Peak relative power output also progressively fell in fast-twitch fibres as Ca2+ activation levels were lowered from maximal down to 50% P4.5. However, in slow-twitch fibres, peak relative power output decreased only at 70% P4.5 and then remained unchanged with further reductions in Ca2+ activation levels. The greater Ca2+ dependence of peak relative power output in cardiac myocytes and fast-twitch fibres may arise from a shared mechanism such as cooperative inactivation of the thin filament, which is likely to be slowest in less cooperative slow-twitch fibres. During submaximal Ca2+ activations, the time course of shortening became markedly curvilinear during isotonic shortening in all three muscle types. The progressive slowdown in shortening velocity during isotonic contractions was greatest in fast-twitch fibres, consistent with the higher degree of cooperativity of Ca2+ activation in fast-twitch fibres. Additionally, fast-twitch and slow-twitch fibre stiffness decreased in concert with the curvature of length traces during loaded shortening. These results are consistent with the idea that cooperative inactivation of the thin filament occurs during loaded shortening and such a mechanism may contribute to the progressive slowing and overall Ca2+ dependence of loaded shortening velocity. PMID- 10811736 TI - Tamoxifen and ATP synergistically activate Cl- release by cultured bovine pigmented ciliary epithelial cells. AB - Purines alter aqueous humour secretion by the bilayered ciliary epithelium. Adenosine but not ATP shrinks non-pigmented ciliary epithelial (NPE) cells by activating Cl- channels. We now report effects of ATP on pigmented ciliary epithelial (PE) cells. Cultured bovine PE cells were studied volumetrically by electronic cell sorting. ATP and tamoxifen acted synergistically to shrink PE cells. Neither ATP nor tamoxifen alone had a consistent effect on cell volume. The tamoxifen, ATP-activated shrinkage required Cl- release since the response was blocked by removing Cl- and was inhibited by the Cl- channel blockers 5-nitro 2-(3-phenylpropylamino)-benzoate and 4,4'-diisothiocyano-2,2'-disulfonic acid. The modulating effect of tamoxifen could have reflected many actions of tamoxifen. Our data do not support the suggestion that tamoxifen inhibits protein kinase C (PKC) or calcium-calmodulin, or that it acts on histamine or carbachol receptors. The shrinkage produced by ATP and tamoxifen was blocked by 17beta oestradiol, but not 17alpha-oestradiol. The cooperative interaction between tamoxifen and ATP was not mediated by an enhanced rise in [Ca2+]i. The results indicate that tamoxifen interacts synergistically with ATP to activate Cl- release by the PE cells. PMID- 10811737 TI - Effects of reduced vesicular filling on synaptic transmission in rat hippocampal neurones. AB - The consequence of reduced uptake of neurotransmitters into synaptic vesicles on synaptic transmission was examined in rat hippocampal slices and culture using bafilomycin A1 (Baf), a potent and specific blocker of the vacuolar-type (V-type) ATPase, which eliminates the driving force for the uptake of both glutamate and GABA into synaptic vesicles. After incubation with Baf, both the amplitude and frequency of GABAergic miniature inhibitory postsynaptic currents (mIPSCs) were reduced in the slice preparation. Similar effects were seen with glutamatergic miniature excitatory postsynaptic currents (mEPSCs) and GABAergic mIPSCs from cultured neurons. This result indicates that vesicular content is reduced by Baf. The dramatic reduction in the frequency of mPSCs could result either from the exocytosis of empty vesicles or from a mechanism which prevents the exocytosis of depleted vesicles. Vesicle cycling was directly examined using confocal imaging with FM 1-43. In the presence of Baf, vesicles could still be endocytosed and they were released at the same probability as from control untreated synapses. Prolonged high-frequency electrical stimulation of synapses in culture failed to alter the amplitude of mEPSCs, suggesting that the filling of vesicles is rapid compared to the rate of vesicle recycling during repetitive synaptic stimulation. Profound release of glutamate with alpha-latrotoxin did cause a small, but reproducible, reduction in quantal size. These results indicate that decreasing the amount of glutamate and GABA in synaptic vesicles reduces quantal size. Furthermore, the probability of vesicle exocytosis appears to be entirely independent of the state of filling of the vesicle. However, even during high frequency action potential-evoked release of glutamate, quantal size remained unchanged. PMID- 10811738 TI - Functional connectivity among ventrolateral medullary respiratory neurones and responses during fictive cough in the cat. AB - This study tested predictions from a network model of ventrolateral medullary respiratory neurone interactions for the generation of the cough motor pattern observed in inspiratory and expiratory pump muscles. Data were from 34 mid collicularly decerebrated, paralysed, artificially ventilated cats. Cough-like patterns (fictive cough) in efferent phrenic and lumbar nerve activities were elicited by mechanical stimulation of the intrathoracic trachea. Neurones in the ventral respiratory group, including the Botzinger and pre-Botzinger complexes, were monitored simultaneously with microelectrode arrays. Spike trains were analysed for evidence of functional connectivity and responses during fictive cough with cycle-triggered histograms, autocorrelograms, cross-correlograms, and spike-triggered averages of phrenic and recurrent laryngeal nerve activities. Significant cross-correlogram features were detected in 151 of 1988 pairs of respiratory modulated neurones. There were 59 central peaks, 5 central troughs, 11 offset peaks and 2 offset troughs among inspiratory neurone pairs. Among expiratory neurones there were 23 central peaks, 8 offset peaks and 4 offset troughs. Correlations between inspiratory and expiratory neurones included 20 central peaks, 10 central troughs and 9 offset troughs. Spike-triggered averages of phrenic motoneurone activity had 51 offset peaks and 5 offset troughs. The concurrent responses and multiple short time scale correlations support parallel and serial network interactions proposed in our model for the generation of the cough motor pattern in the respiratory pump muscles. Inferred associations included the following. (a) Excitation of augmenting inspiratory (I-Aug) neurones and phrenic motoneurones by I-Aug neurones. (b) Inhibition of augmenting expiratory (E-Aug) neurones by decrementing inspiratory (I-Dec) neurones. (c) Inhibition of I-Aug, I-Dec and E-Aug neurones by E-Dec neurones. (d) Inhibition of I-Aug and I-Dec neurones and phrenic motoneurones by E-Aug neurones. The data also confirm previous results and support hypotheses in current network models for the generation of the eupnoeic pattern. PMID- 10811739 TI - Bulbospinal control of spinal cord pathways generating locomotor extensor activities in the cat. AB - Intracellular recording of lumbosacral motoneurones in the decerebrate and partially spinalized cat injected with nialamide and L-dihydroxyphenylalanine (l DOPA) was used to investigate the interneuronal convergence of two bulbospinal pathways and of the segmental pathways involved with the generation of extensor activities during locomotion. Deiter's nucleus (DN) or the medial longitudinal fasciculus (MLF) was stimulated in alternation with, and in combination with, stimulation of group I afferents from extensor muscles or of contralateral flexor reflex afferents (coFRA). The evoked polysynaptic EPSPs were recorded in extensor motoneurones when long-latency, long-lasting discharges were evoked by the stimulation of coFRA and when the group I autogenetic inhibition in extensors was reversed to polysynaptic excitation. Spatial facilitation was inferred when the amplitude of the EPSPs evoked by the combined stimuli was notably larger than the algebraic sum of the EPSPs evoked by individual stimulation. Both DN (16 motoneurones) and MLF inputs (8 motoneurones) showed spatial facilitation when preceded by coFRA stimuli and both could reset the rhythm of fictive stepping by triggering a precocious extensor phase. MLF showed spatial facilitation with extensor group I inputs in 69% of trials but DN failed to show spatial facilitation in any cells. These results indicate that DN and MLF project to the coFRA pathways of the extensor half-centre for locomotion and MLF, but not DN, converge on segmental interneurones of the extensor group I pathways. The implications of such convergence patterns on the functional organization of the extensor half-centre are discussed. PMID- 10811740 TI - Monosynaptic Ia projections from intrinsic hand muscles to forearm motoneurones in humans. AB - Heteronymous Ia excitatory projections from intrinsic hand muscles to human forearm motoneurones (MNs) were investigated. Changes in firing probability of single motor units (MUs) in the flexor carpi radialis (FCR), flexor carpi ulnaris (FCU), flexor digitorum superficialis (FDS), extensor carpi radialis (ECR), extensor carpi ulnaris (ECU) and extensor digitorum communis (EDC) were studied after electrical stimuli were applied to the median and ulnar nerve at wrist level and to the corresponding homonymous nerve at elbow level. Homonymous facilitation, occurring at the same latency as the H reflex, and therefore attributed to monosynaptic Ia EPSPs, was found in all the sampled units. In many MUs an early facilitation was also evoked by heteronymous low-threshold afferents from intrinsic hand muscles. The low threshold (between 0.5 and 0.6 times motor threshold (MT)) and the inability of a pure cutaneous stimulation to reproduce this effect indicate that it is due to stimulation of group I muscle afferents. Evidence for a similar central delay (monosynaptic) in heteronymous as in homonymous pathways was accepted when the difference in latencies of the homonymous and heteronymous peaks did not differ from the estimated supplementary afferent conduction time from wrist to elbow level by more than 0.5 ms (conduction velocity in the fastest Ia afferents between wrist and elbow levels being equal to 69 m s-1). A statistically significant heteronymous monosynaptic Ia excitation from intrinsic hand muscles supplied by both median and ulnar nerves was found in MUs belonging to all forearm motor nuclei tested (although not in ECU MUs after ulnar stimulation). It was, however, more often found in flexors than in extensors, in wrist than in finger muscles and in muscles operating in the radial than in the ulnar side. It is argued that the connections of Ia afferents from intrinsic hand muscles to forearm MNs, which are stronger and more widely distributed than in the cat, might be used to provide a support to the hand during manipulatory movements. PMID- 10811741 TI - Skeletal muscle vasodilatation during sympathoexcitation is not neurally mediated in humans. AB - Evidence for the existence of sympathetic vasodilator nerves in human skeletal muscle is controversial. Manoeuvres such as contralateral ischaemic handgripping to fatigue that cause vasoconstriction in the resting forearm evoke vasodilatation after local alpha-adrenergic receptor blockade, raising the possibility that both constrictor and dilator fibres are present. The purpose of this study was to determine whether this dilatation is neurally mediated. Ten subjects (3 women, 7 men) performed ischaemic handgripping to fatigue before and after acute local anaesthetic block of the sympathetic nerves (stellate ganglion) innervating the contralateral (resting) upper extremity. Forearm blood flow was measured with venous occlusion plethysmography in the resting forearm. In control studies there was forearm vasoconstriction during contralateral handgripping to fatigue. During contralateral handgripping after stellate block, blood flow in the resting forearm increased from 6.1 +/- 0.7 to 18.7 +/- 2.2 ml dl-1 min-1 (P < 0.05). Mean arterial pressure measured concurrently increased from approximately 90 to 130 mmHg and estimated vascular conductance rose from 6.5 +/- 0.7 to 14.0 +/- 1.5 units, indicating that most of the rise in forearm blood flow was due to vasodilatation. Brachial artery administration of beta-blockers (propranolol) and the nitric oxide (NO) synthase inhibitor N G-monomethyl-L-arginine (L-NMMA) after stellate block virtually eliminated all of the vasodilatation to contralateral handgrip. Since vasodilatation was seen after stellate block, our data suggest that sympathetic dilator nerves are not responsible for limb vasodilatation seen during sympathoexcitation evoked by contralateral ischaemic handgripping to fatigue. The results obtained with propranolol and L-NMMA suggest that beta adrenergic mechanisms and local NO release contribute to the dilatation. PMID- 10811742 TI - The influence of heart rate on augmentation index and central arterial pressure in humans. AB - Arterial stiffness is an important determinant of cardiovascular risk. Augmentation index (AIx) is a measure of systemic arterial stiffness derived from the ascending aortic pressure waveform. The aim of the present study was to assess the effect of heart rate on AIx. We elected to use cardiac pacing rather than chronotropic drugs to minimize confounding effects on the systemic circulation and myocardial contractility. Twenty-two subjects (13 male) with a mean age of 63 years and permanent cardiac pacemakers in situ were studied. Pulse wave analysis was used to determine central arterial pressure waveforms, non invasively, during incremental pacing (from 60 to 110 beats min-1), from which AIx and central blood pressure were calculated. Peripheral blood pressure was recorded non-invasively from the brachial artery. There was a significant, inverse, linear relationship between AIx and heart rate (r = -0.76; P < 0.001). For a 10 beats min-1 increment, AIx fell by around 4 %. Ejection duration and heart rate were also inversely related (r = -0. 51; P < 0.001). Peripheral systolic, diastolic and mean arterial pressure increased significantly during incremental pacing. Although central diastolic pressure increased significantly with pacing, central systolic pressure did not. There was a significant increase in the ratio of peripheral to central pulse pressure (P < 0.001), which was accounted for by the observed change in central pressure augmentation. These results demonstrate an inverse, linear relationship between AIx and heart rate. This is likely to be due to alterations in the timing of the reflected pressure wave, produced by changes in the absolute duration of systole. Consideration of wave reflection and aortic pressure augmentation may explain the lack of rise in central systolic pressure during incremental pacing despite an increase in peripheral pressure. PMID- 10811743 TI - Effect of oral glucose on leucine turnover in human subjects at rest and during exercise at two levels of dietary protein. AB - The aim of this study was to determine the effect of glucose supplementation on leucine turnover during and after exercise and whether variation in the previous dietary protein content modulated this effect. Postabsorptive subjects received a primed constant [1-13C, 15N]leucine infusion for 6 h, after previous consumption of a high (1.8 g kg-1 day-1, HP, n = 16) or low (0.7 g kg-1 day-1, LP, n = 16) protein diet for 7 days. The subjects were studied at rest; during 2 h of exercise, during which half of the subjects from each dietary protocol received 0.75 g kg-1 h-1 glucose (HP + G, LP + G) and the other half received water (HP + W, LP + W); then again for 2 h of rest. Glucose supplementation suppressed leucine oxidation (P < 0.01) by 20% in subjects consuming the high protein diet (58.2 +/- 2.8 micromol kg-1 h-1, HP + G; 72.4 +/- 3.9 micromol kg-1 h-1, HP + W) but not the low protein diet (51.1 +/- 5.9 micromol kg-1 h-1, LP + G; 51.7 +/- 5.5 micromol kg-1 h-1, LP + W), with no difference in skeletal muscle branched chain 2-oxo acid dehydrogenase (BCOADH) activity between groups. Glucose supplementation did not alter the rate of whole-body protein synthesis or breakdown. The sparing effect of glucose on leucine oxidation appears only to occur if previous protein intake was high. It was not mediated by a suppression of BCOADH fractional activity but may be due to reduced substrate availability. PMID- 10811744 TI - A 25-year perspective of peripheral nerve surgery: evolving neuroscientific concepts and clinical significance. AB - In spite of an enormous amount of new experimental laboratory data based on evolving neuroscientific concepts during the last 25 years peripheral nerve injuries still belong to the most challenging and difficult surgical reconstructive problems. Our understanding of biological mechanisms regulating posttraumatic nerve regeneration has increased substantially with respect to the role of neurotrophic and neurite-outgrowth promoting substances, but new molecular biological knowledge has so far gained very limited clinical applications. Techniques for clinical approximation of severed nerve ends have reached an optimal technical refinement and new concepts are needed to further increase the results from nerve repair. For bridging gaps in nerve continuity little has changed during the last 25 years. However, evolving principles for immunosuppression may open new perspectives regarding the use of nerve allografts, and various types of tissue engineering combined by bioartificial conduits may also be important. Posttraumatic functional reorganizations occurring in brain cortex are key phenomena explaining much of the inferior functional outcome following nerve repair, and increased knowledge regarding factors involved in brain plasticity may help to further improve the results. Implantation of microchips in the nervous system may provide a new interface between biology and technology and developing gene technology may introduce new possibilities in the manipulation of nerve degeneration and regeneration. PMID- 10811745 TI - Microvascular hand surgery-transplants and replants-over the past 25 years. AB - Two hundred eighty articles related to microvascular transplants and replants published in The Journal of Hand Surgery over the past 25 years were reviewed. Every facet of microsurgery was covered. One hundred seventy-one articles are referenced and discussed under the following major categories: microvascular transplants, replants, nerve grafts, vein grafts, cold intolerance, vasospasm, monitoring, immediate reconstruction, and historical. Contributions were received from 37 countries and 175 authors or groups of authors. PMID- 10811746 TI - The long-term outcome of volar plate arthroplasty of the proximal interphalangeal joint. AB - The ultimate effectiveness of an arthroplasty is most accurately determined by a long-term review of the joints' function and freedom from pain. We examined 17 patients an average of 11.5 years (range, 4-26 years) following volar plate arthroplasty for a fracture-dislocation of the proximal interphalangeal joint. All surgeries were performed by the same surgeon. The average age was 36 years (range, 17-61 years). No patient complained of pain at rest or with activity. Patients having arthroplasty within 4 weeks of injury attained a total active range of motion of 85 degrees (50 degrees to 110 degrees ). Patients having arthroplasty more than 4 weeks after injury (average, 20 weeks) included several elderly women and averaged 61 degrees (30 degrees to 90 degrees ) total active range of motion. Four patients showed some degree of joint narrowing at the follow-up examination. This review suggests that volar plate arthroplasty continues to be of benefit on a long-term basis and provides satisfactory function and pain-free motion. PMID- 10811747 TI - Salvage of failed resection arthroplasties of the distal radioulnar joint using a new ulnar head prosthesis. AB - Twenty-three patients with painful instability following total or partial resection of the ulnar head were treated using a new ulnar head prosthesis combined with a simple soft tissue repair. Stability and marked symptomatic improvement were achieved in all patients. With a mean follow-up period of 27 months, this improvement has been maintained in all but 1 patient, whose prosthesis had to be removed because of a low-grade infection. These results suggest that the use of a prosthesis is a satisfactory way to restore stability and relieve secondary symptoms that may be associated with partial or total excision of the ulnar head. PMID- 10811748 TI - Basal joint arthroplasty using an allograft tendon interposition versus no interposition: a radiographic, vascular, and histologic study. AB - To assess the role of a tendon spacer that fills the trapezial void, the trapeziums were excised and anterior oblique ligaments were reconstructed in 25 monkeys. In addition to the ligament reconstruction, 20 of the monkeys had the trapezial void filled with a tendon allograft. The trapezial space was investigated at 0, 3, 6, 15, and 40 weeks using routine histologic staining, arterial perfusion (Spalteholz), and standardized radiographs. There was a statistically greater decline in trapezial height in the animals without tendon interposition allografts. The tendon grafts became progressively neovascularized and populated with fibroblasts. By 40 weeks, the allograft was no longer a folded tendon but a homogeneous mass of collagen, fibroblasts, and capillaries. The specimens without an interpositional tendon graft had loose fibroadipose tissue filling the carpal void. Polarized light microscopy showed fibers crossing the subchondral bone and moving into the adjacent fibrous spacer in the specimens implanted with a tendon graft. The results indicate that filling the trapezial void with an interposition tendon spacer may aid in maintaining normal wrist anatomy. PMID- 10811749 TI - Thenar insertion of abductor pollicis longus accessory tendons and thumb carpometacarpal osteoarthritis. AB - Although the etiology of osteoarthritis of the thumb carpometacarpal (CMC) joint remains unclear, some theories have focused on variations in the local anatomy of the abductor pollicis longus tendon insertion. This cadaver study of 68 specimens analyzed the relationship between a thenar insertion of an accessory abductor pollicis longus tendon and the presence and severity of thumb CMC osteoarthritis. The joint cartilage surfaces were visually graded for degenerative changes. Thirty-five of 68 specimens (51%) had a thenar insertion, most frequently inserting on either the abductor pollicis brevis or opponens pollicis fascia or muscle belly. No significant association between a thenar insertion and thumb CMC arthritis was observed. Conversely, increasing age was noted to have a significant association with degenerative joint disease. Thus, these findings indicate that a thenar slip of the abductor pollicis longus tendon does not correlate with the presence or severity of CMC osteoarthritis. PMID- 10811750 TI - Peripheral tears of the triangular fibrocartilage complex cause distal radioulnar joint instability after distal radial fractures. AB - The aim of this prospective study was to determine whether peripheral tears of the triangular fibrocartilage complex (TFCC) in patients younger than the osteoporotic age (males,<60 years; females, <50 years) were related to chronic distal radioulnar joint (DRUJ) instability. Fifty-one patients (27 women) with displaced distal radial fractures were included in the study. The median age was 41 years (range, 20-57 years). Arthroscopy at the time of fracture showed complete or partial TFCC tears in 43 patients (24 had only peripheral tears, 10 had only central perforations, and 9 had combined tears). The 1-year (range, 11 27 months) follow-up period included an interview, physical examination, and radiographic evaluation. Ten of the 11 patients with complete peripheral TFCC tears had DRUJ instability at the follow-up examination compared with 7 of the 32 patients with only partial or no peripheral tears. Patients with instability of the DRUJ had a worse Gartland and Werley wrist score. Instability was not associated with any radiographic finding either at the time of fracture or at the follow-up examination. Initial fracture or nonunion of the styloid was even slightly more common in stable patients. PMID- 10811751 TI - Distal radial metaphyseal forces in an extrinsic grip model: implications for postfracture rehabilitation. AB - The purpose of this study was to establish the relationship between force at the distal radius and power grip force of the hand, a common functional and rehabilitation maneuver. This information will provide limits of allowable grip forces during postfixation rehabilitation and guide design requirements for fixation systems. By designing a model of power grip using the extrinsic hand musculotendinous units, we were able to compare grip force with force at the distal radius. Our results show that to obtain 10 N of grip force, approximately 26.3 N of force is transmitted through the distal radius, 52.4 N is transmitted through the radius and ulna combined, and 30.0 N needs to be applied to the flexor tendons. Fifty-one percent of the total forearm force was transmitted through the distal radius in this model. If all forearm forces were transmitted through the radius, 52 N of force would be transmitted through the distal radius to obtain 10 N of grip force. The clinical application of this model suggests that since failure forces of tested distal radius fixation systems range from 55 to 825 N, rehabilitation grip force should not exceed 10 to 159 N, depending on the type of fixation. PMID- 10811752 TI - Self-reported disability following distal radius fractures: the influence of hand dominance. AB - The purpose of this study was to record the spectrum of self-reported disability following distal radius fractures and to gauge for differences in hand dominance in the use of subjective outcome data. Items were generated through patient interviews, literature review, and peer consultation. Fifty-three items were evaluated by a group of 55 patients recovering from a fracture of the distal radius, which established the prevalence, mean severity score, and overall severity score (or impact) of each item as it related to physical function and social/emotional impact. Hand dominance, age, and gender were also recorded. The results confirm that many patients who sustain distal radius fractures experience substantial impairment across a spectrum of quality of life domains. Because patients who sustain a dominant wrist injury are likely to report greater functional impairment across a wider range of activities, they also possess a greater potential for improvement. The practical implication is that outcome studies for the treatment of distal radius fractures should take hand dominance into account. PMID- 10811753 TI - Early excision of heterotopic bone in the forearm. AB - Five patients undergoing early excision of heterotopic bone in the forearm with radiation therapy and indomethacin were reviewed. Inciting causes of heterotopic ossification included distal biceps tendon repairs, ulna fractures, and a fracture of the radius and ulna. Preoperative forearm rotation arc averaged 17 degrees. Excision was performed at an average of 4 months after injury. Patients were administered 500 to 1,000 cGy of radiation after surgery and received indomethacin for 6 weeks. At an average of 37 months after surgery the forearm rotation arc averaged 136 degrees. All patients had resumed normal activities and had no radiographic recurrence of heterotopic bone or other treatment complications. In the past excision of heterotopic ossification after more than 12 months has been advocated to prevent recurrence. The results of this study suggest that early excision with radiation therapy and indomethacin is a viable option in treating and preventing recurrent heterotopic ossification in the forearm. PMID- 10811754 TI - The resistance of a four- and eight-strand suture technique to gap formation during tensile testing: an experimental study of repaired canine flexor tendons after 10 days of in vivo healing. AB - There is a high incidence of gap formation at the repair site following tendon repair. Our goal was to determine the resistance of a 4- and an 8-strand suture technique to gap formation during tensile testing. We hypothesized that the 8 strand repair would sustain higher force levels at the onset of 1- and 3-mm gaps than the 4- strand repair. Twenty-two canine flexor tendons were transected, repaired, and tested to failure after 10 days of in vivo healing. Tests were recorded using a 60-Hz video system that allowed frame-by-frame playback for assessment of gap formation. The 8-strand repairs sustained 80% higher force at a gap of 1 mm than the 4-strand repairs (average force, 70 vs 39 N), but the force sustained at a gap of 3 mm did not differ between groups (35 N for both groups). For both repair types, a 1-mm gap typically occurred near the point of ultimate (maximum) force while a 3-mm gap occurred after the ultimate force. We conclude that the 8-strand repair is significantly more resistant to initial gapping during ex vivo tensile testing than the 4-strand repair but that the two repairs are equally susceptible to rupture if a gap of 3 mm or greater forms. PMID- 10811755 TI - A biomechanical comparison of multistrand flexor tendon repairs using an in situ testing model. AB - An in situ testing model was used to evaluate the performance of zone II flexor tendon repairs and to compare the biomechanical properties of 4-strand repairs with 2- and 6-strand repairs. Fifty digits from human cadaveric hands were mounted in a custom apparatus for in situ tensile testing. Intratendinous metallic markers were placed so that gap formation could be determined by fluoroscopy during tensile testing. Three 4-strand repairs (the 4-strand Kessler, the cruciate, and a locked modification of the cruciate repair) were compared with the 2-strand Kessler and the 6-strand Savage repairs. Ultimate tensile strength, load at 2-, 3-, and 4-mm gap formation, and work of flexion were determined. Work of flexion, while increased for the multistrand repairs, did not show a statistically significant correlation with the number of strands crossing the repair site. The tensile strength of the 6-strand repair was significantly greater than each of the 2- or 4-strand repairs. The tensile strength of all 4 strand repairs was significantly greater than the 2-strand repair. The 6-strand repair and the 2 cruciate repairs demonstrated a statistically increased resistance to gap formation compared with the 2-strand Kessler repair, but notably there was no statistically significant difference in gap resistance between the 2and 4-strand Kessler repairs. This in situ tensiletesting protocol demonstrated that 4- and 6-strand repairs have adequate initial strength to withstand the projected forces of early active motion protocols. Three of the 4 multistrand repairs demonstrated improved gap resistance compared with the 2 strand repair. The presence of the second suture in the Kessler configuration significantly increases its strength but not its gap resistance. PMID- 10811756 TI - Free vascularized bone graft for nonunion of the scaphoid. AB - We used a free vascularized small periosteal bone graft to treat scaphoid nonunion. The graft consisted of periosteum, full-thickness cortex, and the underlying cancellous bone and was harvested from the supracondylar region of the femur. The graft was nourished by the articular branch of the descending geniculate artery and vein. Unlike the currently used vascularized bone grafts, this graft can be easily harvested and shaped to accommodate the bone defect of the scaphoid without disturbing its vascularity and can then be transferred with microvascular anastomosis of the nutrient vessels to the radial artery and its venae commitantes. Ten patients with longstanding nonunion of the scaphoid secondary to avascular necrosis, confirmed by radiologic and intraoperative findings, were treated with this vascularized bone graft. Union was achieved in all 10 patients at an average of 12 weeks after surgery. The average follow-up period for all fractures was 3.5 years. The scores for overall outcome, according to the Mayo wrist scoring system, were excellent in 4 patients, good in 4, and fair in 2. Two unsatisfied patients had preoperative signs of early periscaphoid osteoarthrosis. This free vascularized small bone graft from the supracondylar region of the femur is an attractive alternative to the conventional vascularized bone grafting procedures. PMID- 10811757 TI - Scaphoid nonunions: a 3-dimensional analysis of patterns of deformity. AB - We tested the hypothesis that the fracture location of scaphoid nonunions relates to the fracture displacement, development of dorsal intercalated segment instability (DISI) deformity, and changes in the contact area of the bones in the radiocarpal joint. Eleven patients with scaphoid nonunions were examined with 3 dimensional computed tomography and a new method of proximity mapping. Two different patterns of displacement of scaphoid nonunions were demonstrated, 1 volar and 1 dorsal. All patients with a volar pattern scaphoid nonunion had a DISI deformity. Only a few of the patients with a dorsal pattern scaphoid nonunion, mostly in longstanding nonunions, had a DISI deformity. The fracture line was generally distal to the dorsal apex of the ridge of the scaphoid in the volar-type fractures and proximal in the dorsal displaced fractures. The proximity map of the distal fragment of the scaphoid on the radius in the volar type shifts radial compared with normal; in the distal type it shifts dorsal. Neither of the patterns showed any significant changes of the proximity map in the radiocarpal joint at the proximal scaphoid fragment and the lunate. Whether the fracture line passes distal or proximal to the dorsal apex of the ridge of the scaphoid appears to determine the likelihood of subsequent fracture displacement, DISI deformity, and contact area of the bones in the radiocarpal joint. PMID- 10811758 TI - Reliability of the Lichtman classification of Kienbock's disease. AB - The purpose of this study was to establish the interobserver reliability and intraobserver reproducibility of the staging of Kienbock's disease according to Lichtman's classification. Posteroanterior and lateral wrist radiographs of 64 patients with a diagnosis of Kienbock's disease and 10 control subjects were reviewed independently by 4 observers on 2 separate occasions. The reviewers included 3 hand fellowship-trained surgeons and 1 orthopedist who was not fellowship-trained in hand surgery. A stage was assigned to each set of radiographs according to the Lichtman classification. Paired comparisons for reliability among the 4 observers showed an average absolute percentage agreement of 74% and an average paired weighted kappa coefficient of 0.71. Furthermore, all the controls were correctly classified as stage I, which is in accordance with the Lichtman system. With regard to reproducibility, observers duplicated their initial readings 79% of the time with an average weighted kappa coefficient of 0.77. These results indicate substantial reliability and reproducibility of the Lichtman classification for Kienbock's disease. PMID- 10811759 TI - A model instrument for the documentation of outcome after nerve repair. AB - We present a new model for documentation and quantification of the functional outcome after nerve repair at the wrist or distal forearm level and a protocol that includes a numerical scoring system. The model, presented here along with validation and reliability test results, supports our hypothesis that the summarized test results reflecting specific functional limitations correlates well with the patient's opinion of the impact of the nerve injury on activities of daily living. Seventy patients with nerve repair were examined using the protocol. Analysis included 3 factors: sensory domain (sensory innervation, tactile agnosis, and finger dexterity), motor domain (motor innervation and grip strength), and pain/discomfort domain (hyperaesthesia and cold intolerance). The analysis explained 73% of the variances of the variables and the variables had a logical distribution between the factors. Analysis of internal consistency demonstrated good homogeneity. A calculated total score correlated strongly with the patients' global estimation of the impact of the injury on activities of daily living and the summary of sensory and pain/discomfort domains correlated significantly with the Medical Research Council S0-S4 scale. The presented model represents a useful new tool for evaluation of the functional outcome after nerve injury and repair. PMID- 10811760 TI - Subcutaneous anterior transposition of the ulnar nerve for failed decompression of cubital tunnel syndrome. AB - The current literature universally suggests that submuscular anterior transposition is the standard operative treatment for recurrent cubital tunnel syndrome. Regardless of the type of initial failed procedure, including submuscular transposition, 20 patients underwent anterior subcutaneous transposition of the ulnar nerve. All patients were monitored for a minimum of 2 years after surgery. The most common sites of compression were the medial intermuscular septum and the flexor-pronator aponeurosis. Fifteen patients had a good or excellent outcome; 5 patients had a fair or poor outcome. Relief of pain and paresthesias were the most consistent favorable results. Fair and poor outcomes were significantly associated with increasing age and the number of previous surgeries. Subcutaneous anterior transposition of the ulnar nerve proved to be an effective treatment for recurrent cubital tunnel syndrome. PMID- 10811761 TI - Microvascular skin response to local cooling and body tilt early after digital replantation. AB - To elucidate the alteration in cutaneous microvascular reactivity early after replantation (14-21 days), laser Doppler (LD) flow changes evoked by direct and indirect local cooling and head-up body tilt were studied in the replanted digits (n = 10) and compared with those evoked in the healthy contralateral digits of the same patients (n = 10). During the first 3 minutes of direct cooling of the injured hand, LD flow increased significantly compared with the LD flow before cooling, which confirms that moderate vasodilation is the dominant component of the response to local cooling in skin microcirculation in the early period after replantation. During body tilt, LD flow in the healthy contralateral digit decreased significantly in only the first minute, while LD flow in the replanted digit started to decrease in the second minute after tilting; the decrease was significant from the third to the sixth minute. These results are consistent with the hypothesis that increased human skin alpha-adrenergic receptor sensitivity may be present as early as 2 to 3 weeks after replantation. PMID- 10811762 TI - The use of simultaneous free latissimus dorsi tissue transfers for reconstruction of bilateral upper extremities in a case of purpura fulminans. AB - We report a case of extensive purpura fulminans destroying the soft tissue over the posterior aspect of both elbows. Simultaneous, bilateral free latissimus dorsi muscle transfers were used to close both wounds in a single procedure. The wounds resulting from severe purpura fulminans can be extensive and limb threatening. The simultaneous transfer of 2 free flaps can provide expeditious soft tissue repair while minimizing the risk of repeat anesthesia in these critically ill patients. We found that certain details concerning planning and performing the procedure fostered its successful outcome. PMID- 10811763 TI - Bilateral radial artery pseudoaneurysms associated with bilateral ulnar artery atresia: a case report. AB - Pseudoaneurysms of the radial artery are uncommon and most often localized in an area of penetrating vascular trauma or iatrogenic injury. Hypoplasia of the ulnar artery is even more rare. We report a case of bilateral radial artery pseudoaneurysms associated with complete absence of any ulnar contribution to the vascularity of the hand. A patient presented with bilateral tender masses adjacent to the anatomic snuff boxes that interfered with hand function. After confirming that these masses were bilateral radial artery pseudoaneurysms, resection of the pseudoaneurysms and microscopic reconstruction of the arterial segments preserved vascular integrity of the hands and provided relief of the patient's pain. PMID- 10811764 TI - Subperiosteal ganglion of the distal radius: a case report. AB - We present an unusual case of a subperiosteal ganglion of the distal radius. PMID- 10811765 TI - Extensor pollicis longus tenosynovitis: a case report and review of the literature. AB - A case of tenosynovitis of the extensor pollicis longus, or third dorsal compartment, is presented. This is an unusual condition that may be difficult to diagnose initially. A clinical test for extensor pollicis longus tenosynovitis is described. Operative treatment is recommended. PMID- 10811766 TI - Lipoma arborescens of the elbow: a case report. AB - A 76-year-old woman with a longstanding history of right elbow swelling and recurrent joint effusion presented for consultation. There was no history of trauma and conventional radiographs were negative. Computed tomography (CT) and magnetic resonance imaging were performed and showed a frond-like fatty synovial mass and joint effusion. Arthrotomy disclosed a lipoma arborescens of the right elbow. PMID- 10811767 TI - A new variation of abductor digiti minimi accessorius. AB - An anomalous hypothenar muscle, similar to previous descriptions of an abductor digiti minimi accessorius, was discovered during routine cadaveric dissection. This muscle is unique in that it differs in its insertion from those described previously. In addition, the abductor digiti minimi presented with 2 heads that received innervation from the same ulnar nerve branch as the anomalous muscle rather than from the deep branch of the ulnar nerve. This knowledge is useful to the hand surgeon in diagnosing and surgically treating problems associated with Guyon's canal. PMID- 10811768 TI - 25th anniversary presentation: classic papers in hand surgery. PMID- 10811769 TI - 25th anniversary presentation: classic papers in hand surgery PMID- 10811770 TI - Practice standards in carpal tunnel syndrome electrodiagnosis. PMID- 10811771 TI - Practice standards in carpal tunnel syndrome electrodiagnosis PMID- 10811772 TI - Undereducated, aging, and ... a cycle of decline? PMID- 10811773 TI - Moving research into practice: a new partner. PMID- 10811774 TI - Knowledge-Work for 21st Century Nursing. PMID- 10811775 TI - Guidelines for evidence-based clinical practice. PMID- 10811776 TI - Knowledge development in nursing: our historical roots and future opportunities. PMID- 10811777 TI - The North Carolina Center for Nursing: one state's commitment to planning for nursing resources. PMID- 10811778 TI - Moral reasoning among graduate students in nursing. PMID- 10811779 TI - Pain management in the addicted patient: practical considerations. PMID- 10811781 TI - FAANs to be honored at American Nurses Association's 2000 convention. PMID- 10811780 TI - American Academy of Nursing panel on the future of Medicare. PMID- 10811782 TI - Three AAN Fellows will be inducted into the ANA Hall of Fame. PMID- 10811783 TI - Obituary. PMID- 10811784 TI - Apical control of branch growth and angle in woody plants. AB - Apical control is the inhibition of a lateral branch growth by shoots above it (distal shoots). If the distal shoots are cut off to remove apical control, the lateral branch can grow larger and may bend upwards. Apical control starts when new lateral buds grow after passing through a period of dormancy. Buds initially break and produce leaves, then apical control is exerted and the lower (proximal) laterals stop growing. Apical control also inhibits growth of large, old branches. Gravimorphism and restricted water and nutrient transport can inhibit branch growth, but they are not primary mechanisms of apical control. Apical control may reduce branch photosynthesis. Under apical control allocation of branch-produced assimilate to the stem is relatively high, so low assimilates in the branch may limit branch growth even though hormone levels are adequate for growth. Hormones appear to be involved in apical control, but it is not known how. One role of hormones may be to maintain the strength of the stem sink for branch-produced assimilate. Upward bending of a woody branch after release from apical control requires both new wood production and production of wood cells that can generate an upward bending moment. Apical control inhibits radial growth of branches and, in some species, may regulate the production of wood with an upward bending moment. PMID- 10811785 TI - Evolutionary loss of sepals and/or petals in detarioid legume taxa Aphanocalyx, Brachystegia, and Monopetalanthus (Leguminosae: Caesalpinioideae). AB - Floral development using scanning electron microscopy is compared in several taxa of the Brachystegia subtribal group of caesalpinioid tribe Detariae. This group is characterized by missing sepals and/or petals. In Aphanocalyx djumaensis, Monopetalanthus durandii, and two Brachystegia species, one sepal is initiated in median abaxial position. In the first two, one or two additional sepal rudiments may initiate late. Brachystegia species have all five sepals, which remain scalelike. In Aphanocalyx and Monopetalanthus, one petal initiates adaxially and medianly (a position atypical for the first initiated petal in the family); additional petal rudiments may form in lateral sites. In Brachystegia, five petals are initiated unidirectionally on a meristem ring, but all are suppressed after initiation. In all taxa, ten stamens are initiated on a ring meristem: unidirectionally in Monopetalanthus, bidirectionally in Brachystegia, vs. in erratic order in Aphanocalyx. Carpel and petal initiation are concurrent. Different organ whorls overlap in time in Monopetalanthus and Brachystegia. In all, the floral apex characteristically is elongate radially and narrow tangentially after bracteole initiation. Two ontogenetic features, the meristem ring and the radially elongate post-bracteole floral apex, appear to be possible synapomorphies for the Brachystegia group. PMID- 10811786 TI - Biomechanical analysis of the Rolled (RLD) leaf phenotype of maize. AB - The pleiotropic effects of the Rld1-O/+ mutation of Zea mays (Poaceae) on leaf phenotype include a suppression of normal transverse unrolling, a reversed top/bottom epidermal polarity, and an apparently straighter longitudinal shape. According to engineering shell theory, there might be mechanical coupling between transverse and longitudinal habit, i.e., the leaf rolling itself might produce the longitudinal straightening. We tested this possibility with quantitative curvature measurements and mechanical uncoupling experiments. The contributions of elastic bending under self weight, mechanical coupling, and rest state of leaf parts to the longitudinal and transverse habit were assessed in Rld1-O/+ mutants and a population of sibling +/+ segregants. Elastic bending and curvature coupling are shown to be relatively unimportant. The Rld1-O/+ mutation is shown to alter not only the unrolling process, but also the developmental longitudinal curving in the growing leaf, leading to a straighter midrib and a rolled lamina. The Rld1-O/+ mutant is thus a suitable model to study the relation between tissue polarity and differential curvature development in the maize leaf. Since on the abaxial side of the leaf, more abundant sclerenchyma is found in +/+ than in Rld1 O/+, a gradient in sclerification may contribute to the development of midrib curvature. PMID- 10811787 TI - LEAFY and the evolution of rosette flowering in violet cress (Jonopsidium acaule, Brassicaceae). AB - Arabidopsis and most other Brassicaceae produce an elongated inflorescence of mainly ebracteate flowers. However, the early-flowering species violet cress (Jonopsidium acaule) and a handful of other species produce flowers singly in the axils of rosette leaves. In Arabidopsis the gene LEAFY (LFY) is implicated in both the determination of flower meristem identity and in the suppression of leaves (bracts) that would otherwise subtend the flowers. In this study we examined the role of LFY homologs in the evolution of rosette flowering in violet cress. We cloned two LFY homologs, vcLFY1 and vcLFY2, from violet cress. Their exon sequences show ~90% nucleotide similarity with Arabidopsis LFY and 99% similarity to each other. We used in situ hybridization to study vcLFY expression in violet cress. The patterns were very similar to LFY in Arabidopsis except for stronger expression in the shoot apical meristem outside of the region of flower meristem initiation. It is possible that the relatively diffuse expression of vcLFY contributes to the lack of bract suppression in violet cress. Additionally, the earliest flowers produced by violet cress express vcLFY, suggesting that accelerated flowering in violet cress could also result from changes in the regulation of vcLFY. PMID- 10811788 TI - Megagametophyte development in Potentilla nivea (Rosaceae) from northern Swedish Lapland. AB - An investigation of the embryology of Potentilla nivea was carried out using a confocal laser scanning microscope. The crassinucellate nature of the ovule as well as the presence of a multicellular archesporium was confirmed. A "nucellar cap" builds up as the result of mitotic divisions in the parietal cell tissue and the nucellar epidermis. Earlier reports that several mature megagametophytes are present in each ovule was confirmed. The processes of megasporogenesis and megagametogenesis in Potentilla turned out to differ from descriptions in previous reports, and we propose a reinterpretation. Most importantly, the "archesporium" of previous authors is here considered to be parietal cells, whereas the "chalazal cells" are here interpreted as the archesporium. Meioses commonly occur, giving rise to tetrahedral tetrads. Cytologically unreduced megagametophytes arise from generative tissue (archesporial cells or megasporocytes) only and are diplosporous. Apospory does not occur. The megagametophyte is monosporic and five-nucleate; the three antipodal nuclei normally present in eight-nucleate megagametophytes are missing. PMID- 10811789 TI - Outcrossing rate and inbreeding depression in the perennial yellow bush lupine, Lupinus arboreus (Fabaceae). AB - Little is known about the breeding systems of perennial Lupinus species. We provide information about the breeding system of the perennial yellow bush lupine, Lupinus arboreus, specifically determining self-compatibility, outcrossing rate, and level of inbreeding depression. Flowers are self compatible, but autonomous self-fertilization rarely occurs; thus selfed seed are a product of facilitated selfing. Based on four isozyme loci from 34 maternal progeny arrays of seeds we estimated an outcrossing rate of 0.78. However, when we accounted for differential maturation of selfed seeds, the outcrossing rate at fertilization was lower, ~0.64. Fitness and inbreeding depression of 11 selfed and outcrossed families were measured at four stages: seed maturation, seedling emergence, seedling survivorship, and growth at 12 wk. Cumulative inbreeding depression across all four life stages averaged 0.59, although variation existed between families for the magnitude of inbreeding depression. Inbreeding depression was not manifest uniformly across all four life stages. Outcrossed flowers produced twice as many seeds as selfed flowers, but the mean performance of selfed and outcrossed progeny was not different for emergence, seedling survivorship, and size at 12 wk. Counter to assumptions about this species, L. arboreus is both self-compatible and outcrosses ~78% of the time. PMID- 10811790 TI - Floral dimorphism, pollination, and self-fertilization in gynodioecious GERANIUM RICHARDSONII (Geraniaceae). AB - The selective maintenance of gynodioecy depends on the relative fitness of the male-sterile (female) and hermaphroditic morphs. Females may compensate for their loss of male fitness by reallocating resources from male function (pollen production and pollinator attraction) to female function (seeds and fruits), thus increasing seed production. Females may also benefit from their inability to self fertilize if selfing and inbreeding depression reduce seed quality in hermaphrodites. We investigated how differences in floral resource allocation (flower size) between female and hermaphroditic plants affect two measures of female reproductive success, pollinator visitation and pollen receipt, in gynodioecious populations of Geranium richardsonii in Colorado. Using emasculation treatments in natural populations, we further examined whether selfing by autogamy and geitonogamy comprises a significant proportion of pollen receipt by hermaphrodites. Flowers of female plants are significantly smaller than those of hermaphrodites. The reduction in allocation to pollinator attracting structures (petals) is correlated with a significant reduction in pollinator visitation to female flowers in artificial arrays. The reduction in attractiveness is further manifested in significantly less pollen being deposited on the stigmas of female flowers in natural populations. Autogamy is rare in these protandrous flowers, and geitonogamy accounts for most of the difference in pollen receipt between hermaphrodites and females. Female success at receiving pollen was negatively frequency dependent on the relative frequency of females in populations. Thus, two of the prerequisites for the maintenance of females in gynodioecious populations, differences in resource allocation between floral morphs and high selfing rates in hermaphrodites, occur in G. richardsonii. PMID- 10811791 TI - Pollen dispersal in Yucca filamentosa (Agavaceae): the paradox of self pollination behavior by Tegeticula yuccasella (Prodoxidae). AB - We investigated pollen dispersal in an obligate pollination mutualism between Yucca filamentosa and Tegeticula yuccasella. Yucca moths are the only documented pollinator of yuccas, and moth larvae feed solely on developing yucca seeds. The quality of pollination by a female moth affects larval survival because flowers receiving small amounts of pollen or self-pollen have a high abscission probability, and larvae die in abscised flowers. We tested the prediction that yucca moths primarily perform outcross pollinations by using fluorescent dye to track pollen dispersal in five populations of Y. filamentosa. Dye transfers within plants were common in all populations (mean +/- 1 SE, 55 +/- 3.0%), indicating that moths frequently deposit self-pollen. Distance of dye transfers ranged from 0 to 50 m, and the mean number of flowering plants between the pollen donor and recipient was 5 (median = 0), suggesting that most pollen was transferred among near neighbors. A multilocus genetic estimate of outcrossing based on seedlings matured from open-pollinated fruits at one site was 94 +/- 6% (mean +/- 1 SD). We discuss why moths frequently deposit self-pollen to the detriment of their offspring and compare the yucca-yucca moth interaction with other obligate pollinator mutualisms in which neither pollinator nor plant benefit from self-pollination. PMID- 10811792 TI - Genetic structure and AFLP variation of remnant populations in the rare plant Pedicularis palustris (Scrophulariaceae) and its relation to population size and reproductive components. AB - We investigated plant reproduction in relation to genetic structure, population size, and habitat quality in 13 populations of the rare biennial plant Pedicularis palustris with 3-28500 flowering individuals. We used AFLP (amplified fragment length polymorphism) profiles to analyze genetic similarities among 129 individuals (3-15 per population). In a cluster analysis of genetic similarities most individuals (67%) were arranged in population-specific clusters. Analysis of molecular variance indicated significant genetic differentiation among populations and among and within subpopulations (P < 0.001). Gene flow (N(e) m) was low (0.298). On average, plants produced 55 capsules, 17 seeds per fruit, and 42 seedlings in the following growing season. The number of seeds per capsule was independent of population size and of genetic variability. In contrast, the number of capsules per plant (P < 0.05) and the number of seedlings per plant (P < 0.05) were positively correlated with population size. The relation between population size and the number of seeds per plant was not significant (P = 0.075). The number of capsules and of seeds and seedlings per plant (P < 0.01) were positively correlated with genetic variability. Genetic variability was independent of actual population size, suggesting that historical population processes have to be taken into account, too. Stepwise multiple regressions revealed additional significant relationships of habitat parameters (soil pH, C:N ratio), vegetation composition, and standing crop on reproductive components. We conclude that populations of P. palustris are genetically isolated and that reproductive success most likely is influenced by population size, genetic variability, and habitat quality. Management strategies such as moderate grazing, mowing, and artificial gene flow should endeavor to increase population size as well as genetic variation. PMID- 10811793 TI - Competitive abilities of three narrowly endemic plant species in experimental neighborhoods along a fire gradient. AB - We conducted field experiments manipulating lichens, shrubs, and herbs along a time-since-fire gradient and assessing effects on three endemic herbaceous species of Florida scrub: Eryngium cuneifolium, Hypericum cumulicola, and Polygonella basiramia. Responses included seed germination, survival, biomass, and fecundity. Transplants into recently burned patches generally had higher survival, larger biomass, and greater reproductive output than transplants into long-unburned patches. Open areas and sites near oaks frequently were more favorable than sites near Florida rosemary. Ground lichens did not affect germination but increased mortality rate of seedlings. Neighboring small shrubby and herbaceous species did not affect the performance of these species. Of the three species, naturally occurring E. cuneifolium were farthest from large shrubs, and their microhabitats had the least ground lichens and shrubs. Eryngium cuneifolium and H. cumulicola are capable of forming persistent seed banks and their recruitment after fire depends mostly on these dormant seeds. Polygonella basiramia relies on seed dispersal and immediate seed germination to colonize recently burned patches. Management for these species should involve variable fire regimes to allow all three species to persist along with many other scrub endemics. PMID- 10811794 TI - Photosynthetic and respiratory acclimation and growth response of Antarctic vascular plants to contrasting temperature regimes. AB - Air temperatures have risen over the past 50 yr along the Antarctic Peninsula, and it is unclear what impact this is having on Antarctic plants. We examined the growth response of the Antarctic vascular plants Colobanthus quitensis (Caryophyllaceae) and Deschampsia antarctica (Poaceae) to temperature and also assessed their ability for thermal acclimation, in terms of whole-canopy net photosynthesis (P(n)) and dark respiration (R(d)), by growing plants for 90 d under three contrasting temperature regimes: 7 degrees C day/7 degrees C night, 12 degrees C day/7 degrees C night, and 20 degrees C day/7 degrees C night (18 h/6 h). These daytime temperatures represent suboptimal (7 degrees C), near optimal (12 degrees C), and supraoptimal (20 degrees C) temperatures for P(n) based on field measurements at the collection site near Palmer Station along the west coast of the Antarctic Peninsula. Plants of both species grown at a daytime temperature of 20 degrees C had greater RGR (relative growth rate) and produced 2.2-3.3 times as much total biomass as plants grown at daytime temperatures of 12 degrees or 7 degrees C. Plants grown at 20 degrees C also produced 2.0-4.1 times as many leaves, 3.4-5.5 times as much total leaf area, and had 1.5-1.6 times the LAR (leaf area ratio; leaf area:total biomass) and 1.1-1.4 times the LMR (leaf mass ratio; leaf mass:total biomass) of plants grown at 12 degrees or 7 degrees C. Greater RGR and biomass production at 20 degrees C appeared primarily due to greater biomass allocation to leaf production in these plants. Rates of P(n) (leaf-area basis), when measured at their respective daytime growth temperatures, were highest in plants grown at 12 degrees C, and rates of plants grown at 20 degrees C were only 58 (C. quitensis) or 64% (D. antarctica) of the rates in plants grown at 12 degrees C. Thus, lower P(n) per leaf area in plants grown at 20 degrees C was more than offset by much greater leaf-area production. Rates of whole-canopy P(n) (per plant), when measured at their respective daytime growth temperatures, were highest in plants grown at 20 degrees C, and appeared well correlated with differences in RGR and total biomass among treatments. Colobanthus quitensis exhibited only a slight ability for relative acclimation of P(n) (leaf-area basis) as the optimal temperature for P(n) increased from 8.4 degrees to 10.3 degrees to 11.5 degrees C as daytime growth temperatures increased from 7 degrees to 12 degrees to 20 degrees C. There was no evidence for relative acclimation of P(n) in D. antarctica, as plants grown at all three temperature regimes had a similar optimal temperature (10 degrees C) for P(n). There was no evidence for absolute acclimation of P(n) in either species, as rates of P(n) in plants grown at a daytime temperature of 12 degrees C were higher than those of plants grown at daytime temperatures of 7 degrees or 20 degrees C, when measured at their respective growth temperatures. The poor ability for photosynthetic acclimation in these species may be associated with the relatively stable maritime temperature regime during the growing season along the Peninsula. In contrast to P(n), both species exhibited full acclimation of R(d), and rates of R(d) on a leaf-area basis were similar among treatments when measured at their respective daytime growth temperature. Our results suggest that in the absence of interspecific competition, continued warming along the Peninsula will lead to improved vegetative growth of these species due to (1) greater biomass allocation to leaf-area production (as opposed to improved rates of P(n) per leaf area) and (2) their ability to acclimate R(d), such that respiratory losses per leaf area do not increase under higher temperature regimes. PMID- 10811795 TI - Pollen viability reduction as a potential cost of ant association for Acacia constricta (Fabaceae). AB - Field studies investigating the impact of ants on the reproduction of plants bearing extrafloral nectaries have traditionally focused on seed production, a component of female fitness. The purpose of this study was to test whether ants can affect the pollen viability, a component of male fitness, when they visit flowers of the shrub Acacia constricta. Acacia constricta inflorescences hand pollinated with flowers over which Formica perpilosa ants had crawled set significantly fewer seed pods than inflorescences hand-pollinated by control flowers that had no contact with ants. Many ant species secrete antibiotic substances onto the integument that render pollen inviable, and these secretions are probably the mechanism for reduced pollen viability in this study. The ratio of seed pods produced by self-pollinated inflorescences to those produced by cross-pollinated inflorescences was 0.16, indicating that A. constricta is largely self-incompatible. Because F. perpilosa workers forage primarily on the acacia tree under which they nest, they are unlikely to serve as efficient vectors of outcrossing. Previous work showed that A. constricta shrubs with F. perpilosa ants produce approximately twice as many seeds as similarly sized plants not so associated. The results indicate that association with F. perpilosa could cause a reproductive trade-off for A. constricta: benefits to female function may be accompanied by costs to male function. Selection to discourage ant visitation to flowers may have affected the pollination biology of this and other ant-associated plant species. PMID- 10811796 TI - Phylogeny and biogeography of Eupatorium (Asteraceae: Eupatorieae) based on nuclear ITS sequence data. AB - The classification of the predominantly Neotropical Eupatorieae depends upon the circumscription of the core genus Eupatorium. The recently proposed narrowing of Eupatorium to ~42 species in eastern temperate North America, Europe, and eastern Asia was tested with phylogenetic analysis of nucleotide sequence variation in the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. A total of 40 samples (36 species) of Eupatorieae was analyzed. Several species from North America, South America, and Eurasia that were formerly recognized within a large Eupatorium s.l. (sensu lato) were included in the study. Other taxa included were representative of the majority of the subtribes native to eastern temperate North America. Parsimony analysis supported the contention that Eupatorium be defined narrowly and suggested that Eupatoriadelphus is distinct. The tree topology suggested that Eupatorium and Eupatoriadelphus share a common North American ancestor with Liatris relative to other Eupatorieae. It was apparent that the presumed sister taxa in Eupatoriinae from South America belong to a different clade. These results suggest that, following initial divergence in North America, Eupatorium reached Europe via dispersal during the late Pliocene with subsequent radiation in Asia. PMID- 10811797 TI - Mitochondrial and chloroplast DNA-based phylogeny of Pelargonium (Geraniaceae). AB - Overall phylogenetic relationships within the genus Pelargonium (Geraniaceae) were inferred based on DNA sequences from mitochondrial(mt)-encoded nad1 b/c exons and from chloroplast(cp)-encoded trnL (UAA) 5' exon-trnF (GAA) exon regions using two species of Geranium and Sarcocaulon vanderetiae as outgroups. The group II intron between nad1 exons b and c was found to be absent from the Pelargonium, Geranium, and Sarcocaulon sequences presented here as well as from Erodium, which is the first recorded loss of this intron in angiosperms. Separate phylogenetic analyses of the mtDNA and cpDNA data sets produced largely congruent topologies, indicating linkage between mitochondrial and chloroplast genome inheritance. Simultaneous analysis of the combined data sets yielded a well-resolved topology with high clade support exhibiting a basic split into small and large chromosome species, the first group containing two lineages and the latter three. One large chromosome lineage (x = 11) comprises species from sections Myrrhidium and Chorisma and is sister to a lineage comprising P. mutans (x = 11) and species from section Jenkinsonia (x = 9). Sister to these two lineages is a lineage comprising species from sections Ciconium (x = 9) and Subsucculentia (x = 10). Cladistic evaluation of this pattern suggests that x = 11 is the ancestral basic chromosome number for the genus. PMID- 10811798 TI - Heterochromatin banding patterns in Rutaceae-Aurantioideae--a case of parallel chromosomal evolution. AB - The heterochromatin banding patterns in the karyotypes of 17 species belonging to 15 genera of Rutaceae subfamily Aurantioideae (= Citroideae) were analyzed with the fluorochromes chromomycin (CMA) and 4'-6-diamidino-2-phenylindole-2HCl (DAPI). All species were diploids, except one tetraploid (Clausena excavata) and two hexaploids [Glycosmis parviflora agg. (aggregate) and G. pentaphylla agg.]. There are only CMA/DAPI bands, including those associated with the nucleolus. Using recent cpDNA (chloroplast DNA) sequence data as a phylogenetic background, it becomes evident that generally more basal genera with rather plesiomorphic traits in their morphology, anatomy, and phytochemistry exhibit very small amounts of heterochromatin (e.g., Glycosmis, Severinia, Swinglea), whereas relatively advanced genera from different clades with more apomorphic characters display numerous large CMA bands (e.g., Merrillia, Feroniella, Fortunella). Heterochromatin increase (from 0.7 to 13.7%) is interpreted as apomorphic. The bands are mostly located in the larger chromosomes and at telomeric regions of larger arms. However, one of the largest chromosome pair has been conserved throughout the subfamily with only very little heterochromatin. The heterochromatin-rich patterns observed in different clades of Aurantioideae appear quite similar, suggesting a kind of parallel chromosomal evolution. In respect to the current classification of the subfamily, it is proposed to divide Murraya s.l. (sensu lato) into Bergera and Murraya s.s. (sensu stricto) and to place the former near Clausena into Clauseneae s.s. and the latter together with Merrillia into Citreae s.l. The subtribes recognized within Clauseneae s.s. and Citreae s.l. appear heterogeneous and should be abandoned. On the other hand, the monophyletic nature of the core group of Citrinae, i.e., the Citrus clade with Eremocitrus, Microcitrus, Clymenia, Poncirus, Fortunella, and Citrus, is well supported. PMID- 10811799 TI - Discovery of distyly in Narcissus (Amaryllidaceae). AB - There has been disagreement concerning the nature of sexual polymorphisms in Narcissus, a genus of insect-pollinated geophytes native to the Mediterranean. The existence of trimorphic heterostyly in the genus has recently been confirmed, but the occurrence of distyly remains enigmatic. All sexually dimorphic species previously investigated possess two distinct style lengths but anthers of similar height. Stigma-height dimorphism does not qualify as true distyly because of the absence of a reciprocal correspondence in stigma and anther position in the floral morphs. Such reciprocal herkogamy is generally regarded as the defining feature of heterostyly. Here we report on distyly in N. albimarginatus (section Apodanthae), a rare species confined to a single mountain in northwestern Morocco. A population composed of equal numbers of long- and short-styled plants exhibited reciprocal herkogamy with lower anthers of the long-styled morph and upper anthers of the short-styled morph corresponding in height to stigmas of short- and long-styled plants, respectively. The presence of both stigma-height dimorphism and distyly in Narcissus is of general significance to theoretical models of the evolution of heterostyly. PMID- 10811800 TI - Petrofilaments in palynological preparations. AB - Microscopic structures (petrofilaments) may develop during palynological laboratory processing procedures that are superficially similar to secondary wall thickenings and elaters. These filaments form from angular bodies that resemble fungal spores and resin bodies. The angular bodies are a type of hydrocarbon called asphaltene and extrude the filaments when compounds in the bodies react with solvents in the mounting medium. They may be misinterpreted as fossils and so are illustrated here for the first time in the published literature. PMID- 10811801 TI - The mammalian Mog1 protein is a guanine nucleotide release factor for Ran. AB - Ran is a Ras-related GTPase that is essential for the transport of protein and RNA between the nucleus and the cytoplasm. Proteins that regulate the GTPase cycle and subcellular distribution of Ran include the cytoplasmic GTPase activating protein (RanGAP) and its co-factors (RanBP1, RanBP2), the nuclear guanine nucleotide exchange factor (RanGEF), and the Ran import receptor (NTF2). The recent identification of the Saccharomyces cerevisiae protein Mog1p as a suppressor of temperature-sensitive Ran mutations suggests that additional regulatory proteins remain to be characterized. Here, we describe the identification and biochemical characterization of murine Mog1, which, like its yeast orthologue, is a nuclear protein that binds specifically to RanGTP. We show that Mog1 stimulates the release of GTP from Ran, indicating that Mog1 functions as a guanine nucleotide release factor in vitro. Following GTP release, Mog1 remains bound to nucleotide-free Ran in a conformation that prevents rebinding of the guanine nucleotide. These properties distinguish Mog1 from the well characterized RanGEF and suggest an unanticipated mechanism for modulating nuclear levels of RanGTP. PMID- 10811802 TI - A metal bridge between two enzyme families. 3-deoxy-D-manno-octulosonate-8 phosphate synthase from Aquifex aeolicus requires a divalent metal for activity. AB - The enzymes 3-deoxy-d-manno-octulosonic acid-8-phosphate synthase (KDO8PS) and 3 deoxy-d-arabino-heptulosonic acid-7-phosphate synthase (DAHPS) catalyze analogous condensation reactions between phosphoenolpyruvate and d-arabinose 5-phosphate or d-erythrose 4-phosphate, respectively. While several similarities exist between the two enzymatic reactions, classic studies on the Escherichia coli enzymes have established that DAHPS is a metalloenzyme, whereas KDO8PS has no metal requirement. Here, we demonstrate that KDO8PS from Aquifex aeolicus, representing only the second member of the KDO8PS family to be characterized in detail, is a metalloenzyme. The recombinant KDO8PS, as isolated, displays an absorption band at 505 nm and contains approximately 0.4 and 0.2-0.3 eq of zinc and iron, respectively, per enzyme subunit. EDTA inactivates the enzyme in a time- and concentration-dependent manner and eliminates the absorption at 505 nm. The addition of Cu(2+) to KDO8PS produces an intense absorption at 375 nm, while neither Co(2+) nor Ni(2+) produce such an effect. The EDTA-treated enzyme is reactivated by a wide range of divalent metal ions including Ca(2+), Cd(2+), Co(2+), Cu(2+), Fe(2+), Mg(2+), Mn(2+), Ni(2+), and Zn(2+) and is reversibly inhibited by higher concentrations (>1 mm) of certain metals. Analysis of several metal forms of the enzyme by plasma mass spectrometry suggests that the enzyme preferentially binds one, two, or four metal ions per tetramer. These observations strongly suggest that A. aeolicus KDO8PS is a metalloenzyme in vivo and point to a previously unrecognized relationship between the KDO8PS and DAHPS families. PMID- 10811803 TI - Association of Grb2, Gads, and phospholipase C-gamma 1 with phosphorylated LAT tyrosine residues. Effect of LAT tyrosine mutations on T cell angigen receptor mediated signaling. AB - The linker for activation of T cells (LAT) is a critical adaptor molecule required for T cell antigen receptor (TCR)-mediated signaling and thymocyte development. Upon T cell activation, LAT becomes highly phosphorylated on tyrosine residues, and Grb2, Gads, and phospholipase C (PLC)-gamma1 bind LAT via Src homology-2 domains. In LAT-deficient mutant Jurkat cells, TCR engagement fails to induce ERK activation, Ca(2+) flux, and activation of AP-1 and NF-AT. We mapped the tyrosine residues in LAT responsible for interaction with these specific signaling molecules by expressing LAT mutants with tyrosine to phenylalanine mutations in LAT-deficient cells. Our results showed that three distal tyrosines, Tyr(171), Tyr(191), and Tyr(226), are responsible for Grb2 binding; Tyr(171), and Tyr(191), but not Tyr(226), are necessary for Gads binding. Mutation of Tyr(132) alone abolished PLC-gamma1 binding. Mutation of all three distal tyrosines also abolished PLC-gamma1 binding, suggesting there might be multiple binding sites for PLC-gamma1. Mutation of Tyr(132) affected calcium flux and blocked Erk and NF-AT activation. Since Grb2 binding is not affected by this mutation, these results strongly suggest that PLC-gamma activation regulates Ras activation in these cells. Mutation of individual Grb2 binding sites had no functional effect, but mutation of two or three of these sites, in combination, also affected Erk and NF-AT activation. PMID- 10811805 TI - Function of conserved acidic residues in the PSST homologue of complex I (NADH:ubiquinone oxidoreductase) from Yarrowia lipolytica. AB - Proton-translocating NADH:ubiquinone oxidoreductase (complex I) is the largest and least understood enzyme of the respiratory chain. Complex I from bovine mitochondria consists of more than forty different polypeptides. Subunit PSST has been suggested to carry iron-sulfur center N-2 and has more recently been shown to be involved in inhibitor binding. Due to its pH-dependent midpoint potential, N-2 has been proposed to play a central role both in ubiquinone reduction and proton pumping. To obtain more insight into the functional role of PSST, we have analyzed site-directed mutants of conserved acidic residues in the PSST homologous subunit of the obligate aerobic yeast Yarrowia lipolytica. Mutations D136N and E140Q provided functional evidence that conserved acidic residues in PSST play a central role in the proton translocating mechanism of complex I and also in the interaction with the substrate ubiquinone. When Glu(89), the residue that has been suggested to be the fourth ligand of iron-sulfur center N-2 was changed to glutamine, alanine, or cysteine, the EPR spectrum revealed an unchanged amount of this redox center but was shifted and broadened in the g(z) region. This indicates that Glu(89) is not a ligand of N-2. The results are discussedin the light of structural similarities to the homologous [NiFe] hydrogenases. PMID- 10811804 TI - Substrate recognition domains within extracellular signal-regulated kinase mediate binding and catalytic activation of mitogen-activated protein kinase phosphatase-3. AB - Mitogen-activated protein (MAP) kinase phosphatase-3 (MKP-3) is a dual specificity phosphatase that inactivates extracellular signal-regulated kinase (ERK) MAP kinases. This reflects tight and specific binding between ERK and the MKP-3 amino terminus with consequent phosphatase activation and dephosphorylation of the bound MAP kinase. We have used a series of p38/ERK chimeric molecules to identify domains within ERK necessary for binding and catalytic activation of MKP 3. These studies demonstrate that ERK kinase subdomains V-XI are necessary and sufficient for binding and catalytic activation of MKP-3. These domains constitute the major COOH-terminal structural lobe of ERK. p38/ERK chimeras possessing these regions display increased sensitivity to inactivation by MKP-3. These data also reveal an overlap between ERK domains interacting with MKP-3 and those known to confer substrate specificity on the ERK MAP kinase. Consistent with this, we show that peptides representing docking sites within the target substrates Elk-1 and p90(rsk) inhibit ERK-dependent activation of MKP-3. In addition, abolition of ERK-dependent phosphatase activation following mutation of a putative kinase interaction motif (KIM) within the MKP-3 NH(2) terminus suggests that key sites of contact for the ERK COOH-terminal structural lobe include residues localized between the Cdc25 homology domains (CH2) found conserved between members of the DSP gene family. PMID- 10811806 TI - Dissection of two hallmarks of the open promoter complex by mutation in an RNA polymerase core subunit. AB - Deletion of 10 evolutionarily conserved amino acids from the beta subunit of Escherichia coli RNA polymerase leads to a mutant enzyme that is unable to efficiently hold onto DNA. Open promoter complexes formed by the mutant enzyme are in rapid equilibrium with closed complexes and, unlike the wild-type complexes, are highly sensitive to the DNA competitor heparin (Martin, E., Sagitov, V., Burova, E., Nikiforov, V., and Goldfarb, A. (1992) J. Biol. Chem. 267, 20175-20180). Here we show that despite this instability, the mutant enzyme forms partially open complexes at temperatures as low as 0 degrees C when the wild-type complex is fully closed. Thus, the two hallmarks of the open promoter complex, the stability toward a challenge with DNA competitors and the sensitivity toward low temperature, can be uncoupled by mutation and may be independent in the wild-type complex. We use the high resolution structure of Thermus aquaticus RNA polymerase core to build a functional model of promoter complex formation that accounts for the observed defects of the E. coli RNA polymerase mutants. PMID- 10811807 TI - Insertion and organization within membranes of the delta-endotoxin pore-forming domain, helix 4-loop-helix 5, and inhibition of its activity by a mutant helix 4 peptide. AB - The pore-forming domain of Bacillus thuringiensis Cry1Ac insecticidal protein comprises of a seven alpha-helix bundle (alpha1-alpha7). According to the "umbrella model," alpha4 and alpha5 helices form a hairpin structure thought to be inserted into the membrane upon binding. Here, we have synthesized and characterized the hairpin domain, alpha4-loop-alpha5, its alpha4 and alpha5 helices, as well as mutant alpha4 peptides based on mutations that increased or decreased toxin toxicity. Membrane permeation studies revealed that the alpha4 loop-alpha5 hairpin is extremely active compared with the isolated helices or their mixtures, indicating the complementary role of the two helices and the need for the loop for efficient insertion into membranes. Together with spectrofluorometric studies, we provide direct evidence for the role of alpha4 loop-alpha5 as the membrane-inserted pore-forming hairpin in which alpha4 and alpha5 line the lumen of the channel and alpha5 also participates in the oligomerization of the toxin. Strikingly, the addition of the active alpha4 mutant peptide completely inhibits alpha4-loop-alpha5 pore formation, thus providing, to our knowledge, the first example that a mutated helix within a pore can function as an "immunity protein" by directly interacting with the segments that form the pore. This presents a potential means of interfering with the assembly and function of other membrane proteins as well. PMID- 10811808 TI - Role of asparagine-linked oligosaccharides in rhodopsin maturation and association with its molecular chaperone, NinaA. AB - Many proteins require N-linked glycosylation for conformational maturation and interaction with their molecular chaperones. In Drosophila, rhodopsin (Rh1), the most abundant rhodopsin, is glycosylated in the endoplasmic reticulum (ER) and requires its molecular chaperone, NinaA, for exit from the ER and transport through the secretory pathway. Studies of vertebrate rhodopsins have generated several conflicting proposals regarding the role of glycosylation in rhodopsin maturation. We investigated the role of Rh1 glycosylation and Rh1/NinaA interactions under in vivo conditions by analyzing transgenic flies expressing Rh1 with isoleucine substitutions at each of the two consensus sites for N-linked glycosylation (N20I and N196I). We show that Asn(20) is the sole site for glycosylation. The Rh1(N20I) protein is retained within the secretory pathway, causing an accumulation of ER cisternae and dilation of the Golgi complex. NinaA associates with nonglycosylated Rh1(N20I); therefore, retention of nonglycosylated rhodopsin within the ER is not due to the lack of Rh1(N20I)/NinaA interaction. We further show that Rh1(N20I) interferes with wild type Rh1 maturation and triggers a dominant form of retinal degeneration. We conclude that during maturation Rh1 is present in protein complexes containing NinaA and that Rh1 glycosylation is required for transport of the complexes through the secretory pathway. Failure of this transport process leads to retinal degeneration. PMID- 10811809 TI - A novel system A isoform mediating Na+/neutral amino acid cotransport. AB - A cDNA clone encoding a plasma membrane alanine-preferring transporter (SAT2) has been isolated from glutamatergic neurons in culture and represents the second member of the system A family of neutral amino acid transporters. SAT2 displays a widespread distribution and is expressed in most tissues, including heart, adrenal gland, skeletal muscle, stomach, fat, brain, spinal cord, colon, and lung, with lower levels detected in spleen. No signal is detected in liver or testis. In the central nervous system, SAT2 is expressed in neurons. SAT2 is significantly up-regulated during differentiation of cerebellar granule cells and is absent from astrocytes in primary culture. The functional properties of SAT2, examined using transfected fibroblasts and in cRNA-injected voltage-clamped Xenopus oocytes, show that small aliphatic neutral amino acids are preferred substrates and that transport is voltage- and Na(+)-dependent (1:1 stoichiometry), pH-sensitive, and inhibited by alpha-(methylamino)isobutyric acid (MeAIB), a specific inhibitor of system A. Kinetic analyses of alanine and MeAIB uptake by SAT2 are saturable, with Michaelis constants (K(m)) of 200-500 microm. In addition to its ubiquitous role as a substrate for oxidative metabolism and a major vehicle of nitrogen transport, SAT2 may provide alanine to function as the amino group donor to alpha-ketoglutarate to provide an alternative source for neurotransmitter synthesis in glutamatergic neurons. PMID- 10811810 TI - Role of coatomer and phospholipids in GTPase-activating protein-dependent hydrolysis of GTP by ADP-ribosylation factor-1. AB - The binding of the coat protein complex, coatomer, to the Golgi is mediated by the small GTPase ADP-ribosylation factor-1 (ARF1), whereas the dissociation of coatomer, requires GTP hydrolysis on ARF1, which depends on a GTPase-activating protein (GAP). Recent studies demonstrate that when GAP activity is assayed in a membrane-free environment by employing an amino-terminal truncation mutant of ARF1 (Delta17-ARF1) and a catalytic fragment of the ARF GTPase-activating protein GAP1, GTP hydrolysis is strongly stimulated by coatomer (Goldberg, J., (1999) Cell 96, 893-902). In this study, we investigated the role of coatomer in GTP hydrolysis on ARF1 both in solution and in a phospholipid environment. When GTP hydrolysis was assayed in solution using Delta17-ARF1, coatomer stimulated hydrolysis in the presence of the full-length GAP1 as well as with a Saccharomyces cerevisiae ARF GAP (Gcs1) but had no effect on hydrolysis in the presence of the phosphoinositide dependent GAP, ASAP1. Using wild-type myristoylated ARF1 loaded with GTP in the presence of phospholipid vesicles, GAP1 by itself stimulated GTP hydrolysis efficiently, and coatomer had no additional effect. Disruption of the phospholipid vesicles with detergent resulted in reduced GAP1 activity that was stimulated by coatomer, a pattern that resembled Delta17-ARF1 activity. Our findings suggest that in the biological membrane, the proximity between ARF1 and its GAP, which results from mutual binding to membrane phospholipids, may be sufficient for stimulation of ARF1 GTPase activity. PMID- 10811812 TI - Transcription-induced cleavage of immunoglobulin switch regions by nucleotide excision repair nucleases in vitro. AB - Immunoglobulin (Ig) heavy chain class switch recombination (CSR) mediates isotype switching during B cell development. CSR occurs between switch (S) regions that precede each Ig heavy chain constant region gene. Various studies have demonstrated that transcription plays an essential role in CSR in vivo. In this study, we show that in vitro transcription of S regions in their physiological orientation induces the formation of stable R loops. Furthermore, we show that the nucleotide excision repair nucleases XPF-ERCC1 and XPG can cleave the R loops formed in the S regions. Based on these findings, we propose that CSR is initiated via a mechanism that involves transcription-dependent S region cleavage by DNA structure-specific endonucleases that function in general DNA repair processes. Such a mechanism also may underlie transcription-dependent mutagenic processes such as somatic hypermutation, and contribute to genomic instability in general. PMID- 10811811 TI - Sequence-specific DNA recognition by the Myb-like domain of plant telomeric protein RTBP1. AB - We have identified a rice gene encoding a DNA-binding protein that specifically recognizes the telomeric repeat sequence TTTAGGG found in plants. This gene, which we refer to as RTBP1 (rice telomere-binding protein 1), encodes a polypeptide with a predicted molecular mass of 70 kDa. RTBP1 is ubiquitously expressed in various organs and binds DNA with two or more duplex TTTAGGG repeats. The predicted protein sequence includes a single domain at the C terminus with extensive homology to Myb-like DNA binding motif. The Myb-like domain of RTBP1 is very closely related to that of other telomere-binding proteins, including TRF1, TRF2, Taz1p, and Tbf1p, indicating that DNA-binding domains of telomere-binding proteins are well conserved among evolutionarily distant species. To obtain precise information on the sequence of the DNA binding site recognized by RTBP1, we analyzed the sequence-specific binding properties of the isolated Myb-like domain of RTBP1. The isolated Myb-like domain was capable of sequence-specific DNA binding as a homodimer. Gel retardation analysis with a series of mutated telomere probes revealed that the internal GGGTTT sequence in the two-telomere repeats is critical for binding of Myb-like domain of RTBP1, which is consistent with the model of the TRF1.DNA complex showing that base specific contacts are made within the sequence GGGTTA. To the best of our knowledge, RTBP1 is the first cloned gene in which the product is able to bind double-stranded telomeric DNA in plants. Because the Myb-like domain appears to be a significant motif for a large class of proteins that bind the duplex telomeric DNA, RTBP1 may play important roles in plant telomere function in vivo. PMID- 10811813 TI - Identification of weight-bearing-responsive elements in the skeletal muscle sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA1) gene. AB - The skeletal muscle sarco(endo)plasmic reticulum calcium ATPase (SERCA1) gene is transactivated as early as 2 days after the removal of weight-bearing (Peters, D. G., Mitchell-Felton, H., and Kandarian, S. C. (1999) Am. J. Physiol. 276, C1218 C1225), but the transcriptional mechanisms are elusive. Here, the rat SERCA1 5' flank and promoter region (-3636 to +172 base pairs) was comprehensively examined using in vivo somatic gene transfer into rat soleus muscles (n = 804) to identify region(s) that are both necessary and sufficient for sensitivity to weight bearing. In all, 40 different SERCA1 reporter plasmids were constructed and tested. Several different regions of the SERCA1 5' flank were sufficient to confer a transcriptional response to 7 days of muscle unloading when placed upstream of a heterologous promoter. Two of these regions were analyzed further because they were necessary for the unloading response of -3636 to +172, as demonstrated using internal deletion constructs. Deletion analysis of these regions (-1373 to -1158 and -330 to +172) suggested that unloading responsiveness corresponded to CACC sites and E-boxes. Mutagenesis of cis-elements in the first region showed that a specific CACC box (-1262) was involved in SERCA1 transactivation and a nearby E-box (-1248) was also implicated. Constructs containing trimerized CACC sites and E-boxes showed that the presence of both elements is required to activate transcription. This is the first identification of specific cis-elements required for the regulation of a Ca(2+) handling gene by changes in muscle loading condition. PMID- 10811814 TI - Signaling by covalent heterodimers of interferon-gamma. Evidence for one-sided signaling in the active tetrameric receptor complex. AB - Interferon-gamma (IFN-gamma) and its receptor complex are dimeric and bilaterally symmetric. We created mutants of IFN-gamma that bind only one IFN-gammaR1 chain per dimer molecule (called a monovalent IFN-gamma) to see if the interaction of IFN-gamma with one-half of the receptor complex is sufficient for bioactivity. Mutating a receptor-binding sequence in either AB loop of a covalent dimer of IFN gamma yielded two monovalent IFN-gammas, gamma(m)-gamma and gamma-gamma(m), which cross-link to only a single soluble IFN-gammaR1 molecule in solution and on the cell surface. Monovalent IFN-gamma competes fully with wild type IFN-gamma for binding to U937 cells but only at a greater than 100-fold higher concentration than wild type IFN-gamma. Monovalent IFN-gamma had anti-vesicular stomatitis virus activity and antiproliferative activity, and it induced major histocompatibility complex class I and class II (HLA-DR) expression. In contrast, the maximal levels of activated Stat1alpha produced by monovalent IFN-gammas after 15 min were never more than half of those produced by either wild type or covalent IFN-gammas in human cell lines. These data indicate that while monovalent IFN-gamma activates only one-half of a four-chain receptor complex, this is sufficient for Stat1alpha activation, major histocompatibility complex class I surface antigen induction, and antiviral and antiproliferative activities. Thus, while interaction with both halves of the receptor complex is required for high affinity binding of IFN-gamma and efficient signal transduction, interaction with only one-half of the receptor complex is sufficient to initiate signal transduction. PMID- 10811815 TI - Identification of a third pathway for arachidonic acid mobilization and prostaglandin production in activated P388D1 macrophage-like cells. AB - Previous studies have demonstrated that P388D(1) macrophages are able to mobilize arachidonic acid (AA) and synthesize prostaglandins in two temporally distinct phases. The first phase is triggered by platelet-activating factor within minutes, but needs the cells to be previously exposed to bacterial lipopolysaccharide (LPS) for periods up to 1 h. It is thus a primed immediate phase. The second, delayed phase occurs in response to LPS alone over long incubation periods spanning several hours. Strikingly, the effector enzymes involved in both of these phases are the same, namely the cytosolic group IV phospholipase A(2) (cPLA(2)), the secretory group V phospholipase A(2), and cyclooxygenase-2, although the regulatory mechanisms differ. Here we report that P388D(1) macrophages mobilize AA and produce prostaglandins in response to zymosan particles in a manner that is clearly different from the two described above. Zymosan triggers an immediate AA mobilization response from the macrophages that neither involves the group v phospholipase A(2) nor requires the cells to be primed by LPS. The group VI Ca(2+)-independent phospholipase A(2) is also not involved. Zymosan appears to signal exclusively through activation of the cPLA(2), which is coupled to the cyclooxygenase-2. These results define a secretory PLA(2)-independent pathway for AA mobilization in the P388D(1) macrophages, and demonstrate that, under certain experimental settings, stimulation of the cPLA(2) is sufficient to generate a prostaglandin biosynthetic response in the P388D(1) macrophages. PMID- 10811816 TI - Defining a role and mechanism for IgCAM function in vertebrate axon guidance. PMID- 10811817 TI - EB1 proteins regulate microtubule dynamics, cell polarity, and chromosome stability. PMID- 10811818 TI - Structural transitions at microtubule ends correlate with their dynamic properties in Xenopus egg extracts. AB - Microtubules are dynamically unstable polymers that interconvert stochastically between growing and shrinking states by the addition and loss of subunits from their ends. However, there is little experimental data on the relationship between microtubule end structure and the regulation of dynamic instability. To investigate this relationship, we have modulated dynamic instability in Xenopus egg extracts by adding a catastrophe-promoting factor, Op18/stathmin. Using electron cryomicroscopy, we find that microtubules in cytoplasmic extracts grow by the extension of a two- dimensional sheet of protofilaments, which later closes into a tube. Increasing the catastrophe frequency by the addition of Op18/stathmin decreases both the length and frequency of the occurrence of sheets and increases the number of frayed ends. Interestingly, we also find that more dynamic populations contain more blunt ends, suggesting that these are a metastable intermediate between shrinking and growing microtubules. Our results demonstrate for the first time that microtubule assembly in physiological conditions is a two-dimensional process, and they suggest that the two dimensional sheets stabilize microtubules against catastrophes. We present a model in which the frequency of catastrophes is directly correlated with the structural state of microtubule ends. PMID- 10811819 TI - Oncogenic Ras downregulates Rac activity, which leads to increased Rho activity and epithelial-mesenchymal transition. AB - Proteins of the Rho family regulate cytoskeletal rearrangements in response to receptor stimulation and are involved in the establishment and maintenance of epithelial cell morphology. We recently showed that Rac is able to downregulate Rho activity and that the reciprocal balance between Rac and Rho activity is a major determinant of cellular morphology and motility in NIH3T3 fibroblasts. Using biochemical pull-down assays, we analyzed the effect of transient and sustained oncogenic Ras signaling on the activation state of Rac and Rho in epithelial MDCK cells. In contrast to the activation of Rac by growth factor induced Ras signaling, we found that sustained signaling by oncogenic RasV12 permanently downregulates Rac activity, which leads to upregulation of Rho activity and epithelial-mesenchymal transition. Oncogenic Ras decreases Rac activity through sustained Raf/MAP kinase signaling, which causes transcriptional downregulation of Tiam1, an activator of Rac in epithelial cells. Reconstitution of Rac activity by expression of Tiam1 or RacV12 leads to downregulation of Rho activity and restores an epithelial phenotype in mesenchymal RasV12- or RafCAAX transformed cells. The present data reveal a novel mechanism by which oncogenic Ras is able to interfere with the balance between Rac and Rho activity to achieve morphological transformation of epithelial cells. PMID- 10811820 TI - Intracellular Ca(2+) and Ca(2+)/calmodulin-dependent kinase II mediate acute potentiation of neurotransmitter release by neurotrophin-3. AB - Neurotrophins have been shown to acutely modulate synaptic transmission in a variety of systems, but the underlying signaling mechanisms remain unclear. Here we provide evidence for an unusual mechanism that mediates synaptic potentiation at the neuromuscular junction (NMJ) induced by neurotrophin-3 (NT3), using Xenopus nerve-muscle co-culture. Unlike brain-derived neurotrophic factor (BDNF), which requires Ca(2+) influx for its acute effect, NT3 rapidly enhances spontaneous transmitter release at the developing NMJ even when Ca(2+) influx is completely blocked, suggesting that the NT3 effect is independent of extracellular Ca(2+). Depletion of intracellular Ca(2+) stores, or blockade of inositol 1, 4, 5-trisphosphate (IP3) or ryanodine receptors, prevents the NT3 induced synaptic potentiation. Blockade of IP3 receptors can not prevent BDNF induced potentiation, suggesting that BDNF and NT3 use different mechanisms to potentiate transmitter release. Inhibition of Ca(2+)/calmodulin-dependent kinase II (CaMKII) completely blocks the acute effect of NT3. Furthermore, the NT3 induced potentiation requires a continuous activation of CaMKII, because application of the CaMKII inhibitor KN62 reverses the previously established NT3 effect. Thus, NT3 potentiates neurotransmitter secretion by stimulating Ca(2+) release from intracellular stores through IP3 and/or ryanodine receptors, leading to an activation of CaMKII. PMID- 10811821 TI - Capacitative calcium entry deficits and elevated luminal calcium content in mutant presenilin-1 knockin mice. AB - Dysregulation of calcium signaling has been causally implicated in brain aging and Alzheimer's disease. Mutations in the presenilin genes (PS1, PS2), the leading cause of autosomal dominant familial Alzheimer's disease (FAD), cause highly specific alterations in intracellular calcium signaling pathways that may contribute to the neurodegenerative and pathological lesions of the disease. To elucidate the cellular mechanisms underlying these disturbances, we studied calcium signaling in fibroblasts isolated from mutant PS1 knockin mice. Mutant PS1 knockin cells exhibited a marked potentiation in the amplitude of calcium transients evoked by agonist stimulation. These cells also showed significant impairments in capacitative calcium entry (CCE, also known as store-operated calcium entry), an important cellular signaling pathway wherein depletion of intracellular calcium stores triggers influx of extracellular calcium into the cytosol. Notably, deficits in CCE were evident after agonist stimulation, but not if intracellular calcium stores were completely depleted with thapsigargin. Treatment with ionomycin and thapsigargin revealed that calcium levels within the ER were significantly increased in mutant PS1 knockin cells. Collectively, our findings suggest that the overfilling of calcium stores represents the fundamental cellular defect underlying the alterations in calcium signaling conferred by presenilin mutations. PMID- 10811822 TI - Formation of a complex between nucleolin and replication protein A after cell stress prevents initiation of DNA replication. AB - We used a biochemical screen to identify nucleolin, a key factor in ribosome biogenesis, as a high-affinity binding partner for the heterotrimeric human replication protein A (hRPA). Binding studies in vitro demonstrated that the two proteins physically interact, with nucleolin using an unusual contact with the small hRPA subunit. Nucleolin significantly inhibited both simian virus 40 (SV 40) origin unwinding and SV-40 DNA replication in vitro, likely by nucleolin preventing hRPA from productive interaction with the SV-40 initiation complex. In vivo, use of epifluorescence and confocal microscopy showed that heat shock caused a dramatic redistribution of nucleolin from the nucleolus to the nucleoplasm. Nucleolin relocalization was concomitant with a tenfold increase in nucleolin-hRPA complex formation. The relocalized nucleolin significantly overlapped with the position of hRPA, but only poorly with sites of ongoing DNA synthesis. We suggest that the induced nucleolin-hRPA interaction signifies a novel mechanism that represses chromosomal replication after cell stress. PMID- 10811823 TI - The condensin complex governs chromosome condensation and mitotic transmission of rDNA. AB - We have characterized five genes encoding condensin components in Saccharomyces cerevisiae. All genes are essential for cell viability and encode proteins that form a complex in vivo. We characterized new mutant alleles of the genes encoding the core subunits of this complex, smc2-8 and smc4-1. Both SMC2 and SMC4 are essential for chromosome transmission in anaphase. Mutations in these genes cause defects in establishing condensation of unique (chromosome VIII arm) and repetitive (rDNA) regions of the genome but do not impair sister chromatid cohesion. In vivo localization of Smc4p fused to green fluorescent protein showed that, unexpectedly, in S. cerevisiae the condensin complex concentrates in the rDNA region at the G2/M phase of the cell cycle. rDNA segregation in mitosis is delayed and/or stalled in smc2 and smc4 mutants, compared with separation of pericentromeric and distal arm regions. Mitotic transmission of chromosome III carrying the rDNA translocation is impaired in smc2 and smc4 mutants. Thus, the condensin complex in S. cerevisiae has a specialized function in mitotic segregation of the rDNA locus. Chromatin immunoprecipitation (ChIP) analysis revealed that condensin is physically associated with rDNA in vivo. Thus, the rDNA array is the first identified set of DNA sequences specifically bound by condensin in vivo. The biological role of higher-order chromosome structure in S. cerevisiae is discussed. PMID- 10811824 TI - DNA dendrimers localize MyoD mRNA in presomitic tissues of the chick embryo. AB - MyoD expression is thought to be induced in somites in response to factors released by surrounding tissues; however, reverse transcription-PCR and cell culture analyses indicate that myogenic cells are present in the embryo before somite formation. Fluorescently labeled DNA dendrimers were used to identify MyoD expressing cells in presomitic tissues in vivo. Subpopulations of MyoD positive cells were found in the segmental plate, epiblast, mesoderm, and hypoblast. Directly after laying, the epiblast of the two layered embryo contained approximately 20 MyoD positive cells. These results demonstrate that dendrimers are precise and sensitive reagents for localizing low levels of mRNA in tissue sections and whole embryos, and that cells with myogenic potential are present in the embryo before the initiation of gastrulation. PMID- 10811825 TI - Nuclear import of the ran exchange factor, RCC1, is mediated by at least two distinct mechanisms. AB - RCC1, the only known guanine-nucleotide exchange factor for the Ran GTPase, is an approximately 45-kD nuclear protein that can bind chromatin. An important question concerns how RCC1 traverses the nuclear envelope. We now show that nuclear RCC1 is not exported readily in interphase cells and that the import of RCC1 into the nucleoplasm is extremely rapid. Import can proceed by at least two distinct mechanisms. The first is a classic import pathway mediated by basic residues within the NH(2)-terminal domain (NTD) of RCC1. This pathway is dependent upon both a preexisting Ran gradient and energy, and preferentially uses the importin-alpha3 isoform of importin-alpha. The second pathway is not mediated by the NTD of RCC1. This novel pathway does not require importin-alpha or importin-beta or the addition of any other soluble factor in vitro; however, this pathway is saturable and sensitive only to a subset of inhibitors of classical import pathways. Furthermore, the nuclear import of RCC1 does not require a preexisting Ran gradient or energy. We speculate that this second import pathway evolved to ensure that RCC1 never accumulates in the cytoplasm. PMID- 10811826 TI - Formation of spindle poles by dynein/dynactin-dependent transport of NuMA. AB - NuMA is a large nuclear protein whose relocation to the spindle poles is required for bipolar mitotic spindle assembly. We show here that this process depends on directed NuMA transport toward microtubule minus ends powered by cytoplasmic dynein and its activator dynactin. Upon nuclear envelope breakdown, large cytoplasmic aggregates of green fluorescent protein (GFP)-tagged NuMA stream poleward along spindle fibers in association with the actin-related protein 1 (Arp1) protein of the dynactin complex and cytoplasmic dynein. Immunoprecipitations and gel filtration demonstrate the assembly of a reversible, mitosis-specific complex of NuMA with dynein and dynactin. NuMA transport is required for spindle pole assembly and maintenance, since disruption of the dynactin complex (by increasing the amount of the dynamitin subunit) or dynein function (with an antibody) strongly inhibits NuMA translocation and accumulation and disrupts spindle pole assembly. PMID- 10811827 TI - Microtubule interactions with the cell cortex causing nuclear movements in Saccharomyces cerevisiae. AB - During mitosis in budding yeast the nucleus first moves to the mother-bud neck and then into the neck. Both movements depend on interactions of cytoplasmic microtubules with the cortex. We investigated the mechanism of these movements in living cells using video analysis of GFP-labeled microtubules in wild-type cells and in EB1 and Arp1 mutants, which are defective in the first and second steps, respectively. We found that nuclear movement to the neck is largely mediated by the capture of microtubule ends at one cortical region at the incipient bud site or bud tip, followed by microtubule depolymerization. Efficient microtubule interactions with the capture site require that microtubules be sufficiently long and dynamic to probe the cortex. In contrast, spindle movement into the neck is mediated by microtubule sliding along the bud cortex, which requires dynein and dynactin. Free microtubules can also slide along the cortex of both bud and mother. Capture/shrinkage of microtubule ends also contributes to nuclear movement into the neck and can serve as a backup mechanism to move the nucleus into the neck when microtubule sliding is impaired. Conversely, microtubule sliding can move the nucleus into the neck even when capture/shrinkage is impaired. PMID- 10811828 TI - Zebrafish vasa RNA but not its protein is a component of the germ plasm and segregates asymmetrically before germline specification. AB - Work in different organisms revealed that the vasa gene product is essential for germline specification. Here, we describe the asymmetric segregation of zebrafish vasa RNA, which distinguishes germ cell precursors from somatic cells in cleavage stage embryos. At the late blastula (sphere) stage, vasa mRNA segregation changes from asymmetric to symmetric, a process that precedes primordial germ cell proliferation and perinuclear localization of Vasa protein. Analysis of hybrid fish between Danio rerio and Danio feegradei demonstrates that zygotic vasa transcription is initiated shortly after the loss of unequal vasa mRNA segregation. Blocking DNA replication indicates that the change in vasa RNA segregation is dependent on a maternal program. Asymmetric segregation is impaired in embryos mutant for the maternal effect gene nebel. Furthermore, ultrastructural analysis of vasa RNA particles reveals that vasa RNA, but not Vasa protein, localizes to a subcellular structure that resembles nuage, a germ plasm organelle. The structure is initially associated with the actin cortex, and subsequent aggregation is inhibited by actin depolymerization. Later, the structure is found in close proximity of microtubules. We previously showed that its translocation to the distal furrows is microtubule dependent. We propose that vasa RNA but not Vasa protein is a component of the zebrafish germ plasm. Triggered by maternal signals, the pattern of germ plasm segregation changes, which results in the expression of primordial germ cell-specific genes such as vasa and, consequently, in germline fate commitment. PMID- 10811829 TI - Role of tetanus neurotoxin insensitive vesicle-associated membrane protein (TI VAMP) in vesicular transport mediating neurite outgrowth. AB - How vesicular transport participates in neurite outgrowth is still poorly understood. Neurite outgrowth is not sensitive to tetanus neurotoxin thus does not involve synaptobrevin-mediated vesicular transport to the plasma membrane of neurons. Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI VAMP) is a vesicle-SNARE (soluble N-ethylmaleimide-sensitive fusion protein [NSF] attachment protein [SNAP] receptor), involved in transport to the apical plasma membrane in epithelial cells, a tetanus neurotoxin-resistant pathway. Here we show that TI-VAMP is essential for vesicular transport-mediating neurite outgrowth in staurosporine-differentiated PC12 cells. The NH(2)-terminal domain, which precedes the SNARE motif of TI-VAMP, inhibits the association of TI-VAMP with synaptosome-associated protein of 25 kD (SNAP25). Expression of this domain inhibits neurite outgrowth as potently as Botulinum neurotoxin E, which cleaves SNAP25. In contrast, expression of the NH(2)-terminal deletion mutant of TI-VAMP increases SNARE complex formation and strongly stimulates neurite outgrowth. These results provide the first functional evidence for the role of TI-VAMP in neurite outgrowth and point to its NH(2)-terminal domain as a key regulator in this process. PMID- 10811830 TI - Distinct membrane domains on endosomes in the recycling pathway visualized by multicolor imaging of Rab4, Rab5, and Rab11. AB - Two endosome populations involved in recycling of membranes and receptors to the plasma membrane have been described, the early and the recycling endosome. However, this distinction is mainly based on the flow of cargo molecules and the spatial distribution of these membranes within the cell. To get insights into the membrane organization of the recycling pathway, we have studied Rab4, Rab5, and Rab11, three regulatory components of the transport machinery. Following transferrin as cargo molecule and GFP-tagged Rab proteins we could show that cargo moves through distinct domains on endosomes. These domains are occupied by different Rab proteins, revealing compartmentalization within the same continuous membrane. Endosomes are comprised of multiple combinations of Rab4, Rab5, and Rab11 domains that are dynamic but do not significantly intermix over time. Three major populations were observed: one that contains only Rab5, a second with Rab4 and Rab5, and a third containing Rab4 and Rab11. These membrane domains display differential pharmacological sensitivity, reflecting their biochemical and functional diversity. We propose that endosomes are organized as a mosaic of different Rab domains created through the recruitment of specific effector proteins, which cooperatively act to generate a restricted environment on the membrane. PMID- 10811831 TI - Caenorhabditis elegans beta-G spectrin is dispensable for establishment of epithelial polarity, but essential for muscular and neuronal function. AB - The Caenorhabditis elegans genome encodes one alpha spectrin subunit, a beta spectrin subunit (beta-G), and a beta-H spectrin subunit. Our experiments show that the phenotype resulting from the loss of the C. elegans alpha spectrin is reproduced by tandem depletion of both beta-G and beta-H spectrins. We propose that alpha spectrin combines with the beta-G and beta-H subunits to form alpha/beta-G and alpha/beta-H heteromers that perform the entire repertoire of spectrin function in the nematode. The expression patterns of nematode beta-G spectrin and vertebrate beta spectrins exhibit three striking parallels including: (1) beta spectrins are associated with the sites of cell-cell contact in epithelial tissues; (2) the highest levels of beta-G spectrin occur in the nervous system; and (3) beta spectrin-G in striated muscle is associated with points of attachment of the myofilament apparatus to adjacent cells. Nematode beta-G spectrin associates with plasma membranes at sites of cell-cell contact, beginning at the two-cell stage, and with a dramatic increase in intensity after gastrulation when most cell proliferation has been completed. Strikingly, depletion of nematode beta-G spectrin by RNA-mediated interference to undetectable levels does not affect the establishment of structural and functional polarity in epidermis and intestine. Contrary to recent speculation, beta-G spectrin is not associated with internal membranes and depletion of beta-G spectrin was not associated with any detectable defects in secretion. Instead beta-G spectrin-deficient nematodes arrest as early larvae with progressive defects in the musculature and nervous system. Therefore, C. elegans beta-G spectrin is required for normal muscle and neuron function, but is dispensable for embryonic elongation and establishment of early epithelial polarity. We hypothesize that heteromeric spectrin evolved in metazoans in response to the needs of cells in the context of mechanically integrated tissues that can withstand the rigors imposed by an active organism. PMID- 10811832 TI - Mutations in beta-spectrin disrupt axon outgrowth and sarcomere structure. AB - beta-Spectrin is a major component of the membrane skeleton, a structure found at the plasma membrane of most animal cells. beta-Spectrin and the membrane skeleton have been proposed to stabilize cell membranes, generate cell polarity, or localize specific membrane proteins. We demonstrate that the Caenorhabditis elegans homologue of beta-spectrin is encoded by the unc-70 gene. unc-70 null mutants develop slowly, and the adults are paralyzed and dumpy. However, the membrane integrity is not impaired in unc-70 animals, nor is cell polarity affected. Thus, beta-spectrin is not essential for general membrane integrity or for cell polarity. However, beta-spectrin is required for a subset of processes at cell membranes. In neurons, the loss of beta-spectrin leads to abnormal axon outgrowth. In muscles, a loss of beta-spectrin leads to disorganization of the myofilament lattice, discontinuities in the dense bodies, and a reduction or loss of the sarcoplasmic reticulum. These defects are consistent with beta-spectrin function in anchoring proteins at cell membranes. PMID- 10811833 TI - Inhibition of cytokinesis by a lipid metabolite, psychosine. AB - Although a number of cellular components of cytokinesis have been identified, little is known about the detailed mechanisms underlying this process. Here, we report that the lipid metabolite psychosine (galactosylsphingosine), derived from galactosylceramide, induced formation of multinuclear cells from a variety of nonadherent and adherent cells due to inhibition of cytokinesis. When psychosine was added to the human myelomonocyte cell line U937, which was the most sensitive among the cell lines tested, cleavage furrow formed either incompletely or almost completely. However, abnormal contractile movement was detected in which the cellular contents of one of the hemispheres of the contracting cell were transferred into its counterpart. Finally, the cleavage furrow disappeared and cytokinesis was reversed. Psychosine treatment also induced giant clots of actin filaments in the cells that probably consisted of small vacuoles with filamentous structures, suggesting that psychosine affected actin reorganization. These observations could account for the formation of multinuclear globoid cells in the brains of patients with globoid cell leukodystrophy, a neurological disorder characterized by the accumulation of psychosine due to galactosylceramidase deficiency. PMID- 10811834 TI - A direct interaction of axonin-1 with NgCAM-related cell adhesion molecule (NrCAM) results in guidance, but not growth of commissural axons. AB - An interaction of growth cone axonin-1 with the floor-plate NgCAM-related cell adhesion molecule (NrCAM) was shown to play a crucial role in commissural axon guidance across the midline of the spinal cord. We now provide evidence that axonin-1 mediates a guidance signal without promoting axon elongation. In an in vitro assay, commissural axons grew preferentially on stripes coated with a mixture of NrCAM and NgCAM. This preference was abolished in the presence of anti axonin-1 antibodies without a decrease in neurite length. Consistent with these findings, commissural axons in vivo only fail to extend along the longitudinal axis when both NrCAM and NgCAM interactions, but not when axonin-1 and NrCAM or axonin-1 and NgCAM interactions, are perturbed. Thus, we conclude that axonin-1 is involved in guidance of commissural axons without promoting their growth. PMID- 10811835 TI - The tetraspan molecule CD151, a novel constituent of hemidesmosomes, associates with the integrin alpha6beta4 and may regulate the spatial organization of hemidesmosomes. AB - CD151 is a cell surface protein that belongs to the tetraspan superfamily. It associates with other tetraspan molecules and certain integrins to form large complexes at the cell surface. CD151 is expressed by a variety of epithelia and mesenchymal cells. We demonstrate here that in human skin CD151 is codistributed with alpha3beta1 and alpha6beta4 at the basolateral surface of basal keratinocytes. Immunoelectron microscopy showed that CD151 is concentrated in hemidesmosomes. By immunoprecipitation from transfected K562 cells, we established that CD151 associates with alpha3beta1 and alpha6beta4. In beta4 deficient pyloric atresia associated with junctional epidermolysis bullosa (PA JEB) keratinocytes, CD151 and alpha3beta1 are clustered together at the basal cell surface in association with patches of laminin-5. Focal adhesions are present at the periphery of these clusters, connected with actin filaments, and they contain both CD151 and alpha3beta1. Transient transfection studies of PA-JEB cells with beta4 revealed that the integrin alpha6beta4 becomes incorporated into the alpha3beta1-CD151 clusters where it induces the formation of hemidesmosomes. As a result, the amount of alpha3beta1 in the clusters diminishes and the protein becomes restricted to the peripheral focal adhesions. Furthermore, CD151 becomes predominantly associated with alpha6beta4 in hemidesmosomes, whereas its codistribution with alpha3beta1 in focal adhesions becomes partial. The localization of alpha6beta4 in the pre-hemidesmosomal clusters is accompanied by a strong upregulation of CD151, which is at least partly due to increased cell surface expression. Using beta4 chimeras containing the extracellular and transmembrane domain of the IL-2 receptor and the cytoplasmic domain of beta4, we found that for recruitment of CD151 into hemidesmosomes, the beta4 subunit must be associated with alpha6, confirming that integrins associate with tetraspans via their alpha subunits. CD151 is the only tetraspan identified in hemidesmosomal structures. Others, such as CD9 and CD81, remain diffusely distributed at the cell surface. In conclusion, we show that CD151 is a major component of (pre)-hemidesmosomal structures and that its recruitment into hemidesmosomes is regulated by the integrin alpha6beta4. We suggest that CD151 plays a role in the formation and stability of hemidesmosomes by providing a framework for the spatial organization of the different hemidesmosomal components. PMID- 10811836 TI - Growth plate compressions and altered hematopoiesis in collagen X null mice. AB - A variable skeleto-hematopoietic phenotype was observed in collagen X null mice which mirrored the defects in transgenic (Tg) mice with dominant interference collagen X mutations (Jacenko, O., P. LuValle, and B.R. Olsen. 1993. Nature. 365:56-61). Specifically, perinatal lethality was seen in approximately 10.8% of null mutants at week three after birth, and in another subset by 12 wk. In perinatal lethal mutants, growth plates were compressed, trabecular bone reduced, and hematopoietic aplasia and erythrocyte-filled vascular sinusoids were apparent in marrows. Lymphatic organs, reduced to approximately 80% that of controls, displayed altered architecture and lymphocyte content. In thymuses, a paucity of cortical CD3(+)/CD4(+)/CD8(+) lymphocytes was consistent with the marrow's inability to replenish maturing T cells. In spleens, an unaltered T cell distribution was coupled with diffuse staining for IgD(+)/B220(+) B cells, whose reduction was prominent in poorly organized lymphatic nodules. Disorderly arrays of splenic macrophages surrounding periarteriolar lymphatic sheaths and a red pulp depletion further complemented the Tg perinatal lethal phenotype. Moreover, subtle growth plate compressions and hematopoietic changes were seen in all null mice. Data from Tg and null mice implicate the disruption of collagen X function in the observed skeleto-hematopoietic defects, and suggest that hypertrophic cartilage and endochondral skeletogenesis may contribute to the marrow microenvironment prerequisite for blood cell differentiation. PMID- 10811837 TI - Eosinophil-dependent bromination in the pathogenesis of asthma. PMID- 10811838 TI - Proving HIV-1 immunity: new tools offer new opportunities. PMID- 10811839 TI - The new kids on the block: ADAMTSs, potentially multifunctional metalloproteinases of the ADAM family. PMID- 10811840 TI - Meeting Koch's postulates for calcium signaling in cardiac hypertrophy. PMID- 10811841 TI - Airway plumbing. PMID- 10811842 TI - ADAMTS-1: a metalloproteinase-disintegrin essential for normal growth, fertility, and organ morphology and function. AB - A disintegrin and metalloproteinase (ADAM) represents a protein family possessing both metalloproteinase and disintegrin domains. ADAMTS-1, an ADAM family member cloned from cachexigenic colon adenocarcinoma, is unusual in that it contains thrombospondin type I motifs and anchors to the extracellular matrix. To elucidate the biological role of ADAMTS-1, we developed ADAMTS-1-null mice by gene targeting. Targeted disruption of the mouse ADAMTS-1 gene resulted in growth retardation with adipose tissue malformation. Impaired female fertilization accompanied by histological changes in the uterus and ovaries also resulted. Furthermore, ADAMTS-1(-/-) mice demonstrated enlarged renal calices with fibrotic changes from the ureteropelvic junction through the ureter, and abnormal adrenal medullary architecture without capillary formation. ADAMTS-1 thus appears necessary for normal growth, fertility, and organ morphology and function. Moreover, the resemblance of the renal phenotype to human ureteropelvic junction obstruction may provide a clue to the pathogenesis of this common congenital disease. PMID- 10811843 TI - Cubilin is an albumin binding protein important for renal tubular albumin reabsorption. AB - Using affinity chromatography and surface plasmon resonance analysis, we have identified cubilin, a 460-kDa receptor heavily expressed in kidney proximal tubule epithelial cells, as an albumin binding protein. Dogs with a functional defect in cubilin excrete large amounts of albumin in combination with virtually abolished proximal tubule reabsorption, showing the critical role for cubilin in the uptake of albumin by the proximal tubule. Also, by immunoblotting and immunocytochemistry we show that previously identified low-molecular-weight renal albumin binding proteins are fragments of cubilin. In addition, we find that mice lacking the endocytic receptor megalin show altered urinary excretion, and reduced tubular reabsorption, of albumin. Because cubilin has been shown to colocalize and interact with megalin, we propose a mechanism of albumin reabsorption mediated by both of these proteins. This process may prove important for understanding interstitial renal inflammation and fibrosis caused by proximal tubule uptake of an increased load of filtered albumin. PMID- 10811844 TI - Predominant role for C5b-9 in renal ischemia/reperfusion injury. AB - Previous work has indicated that complement is a mediator of ischemia/reperfusion (I/R) injury. To investigate the components of complement responsible for this effect, we examined a model of renal I/R injury in C3-, C4-, C5-, and C6 deficient mice. We occluded the renal arteries and veins (40-58 minutes) and, after reperfusion (0-72 hours), assessed renal structural and functional injury. C3-, C5-, and C6-deficient mice were protected from renal I/R injury, whereas C4 deficient mice were not protected. C6-deficient mice treated with antibody to block C5a generation showed no additional protection from I/R injury. Reconstitution with C6 alone restored the I/R injury in C6-deficient mice. Tubular epithelial cells were the main structures damaged by complement-mediated attack, and, in contrast, the renal vessels were spared. Neutrophil infiltration and myeloperoxidase activity were reduced in C-deficient mouse kidney, but by a similar extent in C3-deficient and C6-deficient mice. We conclude that the membrane attack complex of complement (in which C5 and C6 participate) may account for the effect of complement on mouse renal I/R injury. Neither C5a mediated neutrophil infiltration nor the classic pathway, in which C4 participates, appears to contribute to I/R injury in this model. By contrast with other organs, such as the heart, the primary effect of complement in the ischemic area is on the parenchymal cell rather than the vascular endothelial cell. The membrane attack complex of complement is a potential target for prevention of I/R injury in this model. PMID- 10811845 TI - Disruption of ECE-1 and ECE-2 reveals a role for endothelin-converting enzyme-2 in murine cardiac development. AB - Endothelin-converting enzyme-1 and -2 (ECE-1 and -2) are membrane-bound metalloproteases that can cleave biologically the inactive endothelin-1 (ET-1) precursor to form active ET-1 in vitro. We previously reported developmental defects in specific subsets of neural crest-derived tissues, including branchial arch-derived craniofacial structures, aortic arch arteries, and the cardiac outflow tract in ECE-1 knockout mice. To examine the role of ECE-2 in cardiovascular development, we have now generated a null mutation in ECE-2 by homologous recombination. ECE-2 null mice develop normally, are healthy into adulthood, are fertile in both sexes, and live a normal life span. However, when they are bred into an ECE-1-null background, defects in cardiac outflow structures become more severe than those in ECE-1 single knockout embryos. In addition, ECE-1(-/-); ECE-2(-/-) double null embryos exhibited abnormal atrioventricular valve formation, a phenotype never seen in ECE-1 single knockout embryos. In the developing mouse heart, ECE-2 mRNA is expressed in the endocardial cushion mesenchyme from embyronic day (E) 12.5, in contrast to the endocardial expression of ECE-1. Levels of mature ET-1 and ET-2 in whole ECE-1(-/ ); ECE-2(-/-) embryos at E12.5 do not differ appreciably from those of ECE-1(-/-) embryos. The significant residual ET-1/ET-2 in the ECE-1(-/-); ECE-2(-/-) embryos indicates that proteases distinct from ECE-1 and ECE-2 can carry out ET-1 activation in vivo. PMID- 10811846 TI - Macrophage inflammatory protein 3alpha transgene attracts dendritic cells to established murine tumors and suppresses tumor growth. AB - Dendritic cells (DCs) are powerful antigen-presenting cells that function as the principal activators of T cells. Since the human CC chemokine, macrophage inflammatory protein 3alpha (MIP-3alpha), is chemotactic for DCs in vitro, we hypothesized that adenovirus-mediated gene transfer of MIP-3alpha (AdMIP-3alpha) to tumors might induce local accumulation of DCs and inhibit growth of preexisting tumors. AdMIP-3alpha directed expression of mRNA and protein in vitro, and the supernatant of A549 cells infected with AdMIP-3alpha was chemotactic for DCs. In vivo, injection of AdMIP-3alpha into subcutaneous tumors resulted in local expression of the MIP-3alpha cDNA and in the local accumulation of DCs. In four syngeneic tumor models, growth of established tumors was significantly inhibited compared with untreated tumors or tumors injected with control vector, and in all but the poorly immunogenic LLC carcinoma model, this treatment increased survival advantage of the preexisting tumors. In all four tumor models, intratumoral injection of AdMIP-3alpha induced the local accumulation of CD8b. 2(+) cells and elicited tumor-specific cytotoxic T lymphocyte activity, and adoptive transfer of splenocytes of animals receiving this treatment protected against a subsequent challenge with the identical tumor cells. In wild-type but not in CD8-deficient mice, AdMIP-3alpha inhibited the growth of tumors. Finally, AdMIP-3alpha also inhibited the growth of distant tumors. This strategy may be useful for enlisting the help of DCs to boost anti tumor immunity against local and metastatic tumors without the necessity of ex vivo isolation and manipulation of DCs. PMID- 10811847 TI - CaM kinase signaling induces cardiac hypertrophy and activates the MEF2 transcription factor in vivo. AB - Hypertrophic growth is an adaptive response of the heart to diverse pathological stimuli and is characterized by cardiomyocyte enlargement, sarcomere assembly, and activation of a fetal program of cardiac gene expression. A variety of Ca(2+) dependent signal transduction pathways have been implicated in cardiac hypertrophy, but whether these pathways are independent or interdependent and whether there is specificity among them are unclear. Previously, we showed that activation of the Ca(2+)/calmodulin-dependent protein phosphatase calcineurin or its target transcription factor NFAT3 was sufficient to evoke myocardial hypertrophy in vivo. Here, we show that activated Ca(2+)/calmodulin-dependent protein kinases-I and -IV (CaMKI and CaMKIV) also induce hypertrophic responses in cardiomyocytes in vitro and that CaMKIV overexpressing mice develop cardiac hypertrophy with increased left ventricular end-diastolic diameter and decreased fractional shortening. Crossing this transgenic line with mice expressing a constitutively activated form of NFAT3 revealed synergy between these signaling pathways. We further show that CaMKIV activates the transcription factor MEF2 through a posttranslational mechanism in the hypertrophic heart in vivo. Activated calcineurin is a less efficient activator of MEF2-dependent transcription, suggesting that the calcineurin/NFAT and CaMK/MEF2 pathways act in parallel. These findings identify MEF2 as a downstream target for CaMK signaling in the hypertrophic heart and suggest that the CaMK and calcineurin pathways preferentially target different transcription factors to induce cardiac hypertrophy. PMID- 10811848 TI - HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death. AB - We have tracked the in vivo migration and have identified in vivo correlates of cytotoxic T-lymphocyte (CTL) activity in HIV-seropositive subjects infused with autologous gene-marked CD8(+) HIV-specific CTL. The number of circulating gene marked CTL ranged from 1.6 to 3.5% shortly after infusion to less than 0.5% 2 weeks later. Gene-marked CTL were present in the lymph node at 4.5- to 11-fold excess and colocalized within parafollicular regions of the lymph node adjacent to cells expressing HIV tat fusion transcripts, a correlate of virus replication. The CTL clones expressed the CCR5 receptor and localized among HIV-infected cells expressing the ligands MIP-1alpha and MIP-1beta, CC-chemokines produced at sites of virus replication. Aggregates of apoptotic cells and cells expressing granzyme B localized within these same sites. In contrast, lymph node sections from untreated HIV-seropositive subjects, all with significant viral burden (> 50,000 HIV RNA copies/mL plasma), showed no CC-chemokine expression and exhibited only sporadic and randomly distributed cells expressing granzymes and/or apoptotic cells. These studies show that the infused CTL specifically migrate to sites of HIV replication and retain their antigen-specific cytolytic potential. Moreover, these studies provide a methodology that will facilitate studies of both the magnitude and functional phenotype of Ag-specific CD8(+) T cells in vivo. PMID- 10811849 TI - Osmotic water permeabilities of cultured, well-differentiated normal and cystic fibrosis airway epithelia. AB - Current hypotheses describing the function of normal airway surface liquid (ASL) in lung defense are divergent. One theory predicts that normal airways regulate ASL volume by modulating the flow of isosmotic fluid across the epithelium, whereas an alternative theory predicts that ASL is normally hyposmotic. These hypotheses predict different values for the osmotic water permeability (P(f)) of airway epithelia. We measured P(f) of cultures of normal and cystic fibrosis (CF) airway epithelia that, like the native tissue, contain columnar cells facing the lumen and basal cells that face a basement membrane. Xz laser scanning confocal microscopy recorded changes in epithelial height and transepithelial volume flow in response to anisosmotic challenges. With luminal hyperosmotic challenges, transepithelial and apical membrane P(f) are relatively high for both normal and CF airway epithelia, consistent with an isosmotic ASL. Simultaneous measurements of epithelial cell volume and transepithelial water flow revealed that airway columnar epithelial cells behave as osmometers whose volume is controlled by luminal osmolality. Basal cell volume did not change in these experiments. When the serosal side of the epithelium was challenged with hyperosmotic solutions, the basal cells shrank, whereas the lumen-facing columnar cells did not. We conclude that (a) normal and CF airway epithelia have relatively high water permeabilities, consistent with the isosmotic ASL theory, and the capacity to restore water on airway surfaces lost by evaporation, and (b) the columnar cell basolateral membrane and tight junctions limit transepithelial water flow in this tissue. PMID- 10811850 TI - In a novel form of IFN-gamma receptor 1 deficiency, cell surface receptors fail to bind IFN-gamma. AB - Complete IFN-gamma receptor ligand-binding chain (IFNgammaR1) deficiency is a life-threatening autosomal recessive immune disorder. Affected children invariably die of mycobacterial infection, unless bone marrow transplantation is undertaken. Pathogenic IFNGR1 mutations identified to date include nonsense and splice mutations and frameshift deletions and insertions. All result in a premature stop codon upstream from the segment encoding the transmembrane domain, precluding cell surface expression of the receptors. We report herein two sporadic and two familial cases of a novel form of complete IFNgammaR1 deficiency in which normal numbers of receptors are detected at the cell surface. Two in frame deletions and two missense IFNGR1 mutations were identified in the segment encoding the extracellular ligand-binding domain of the receptor. Eight independent IFNgammaR1-specific mAb's, including seven blocking antibodies, gave recognition patterns that differed between patients, suggesting that different epitopes were altered by the mutations. No specific binding of (125)I-IFN-gamma to cells was observed in any patient, however, and the cells failed to respond to IFN-gamma. The mutations therefore cause complete IFNgammaR1 deficiency by disrupting the IFN-gamma-binding site without affecting surface expression. The detection of surface IFNgammaR1 molecules by specific antibodies, including blocking antibodies, does not exclude a diagnosis of complete IFNgammaR1 deficiency. PMID- 10811851 TI - Restored insulin-sensitivity in IRS-1-deficient mice treated by adenovirus mediated gene therapy. AB - Insulin resistance is commonly observed both in overt diabetes and in individuals prone to, but not yet manifesting, diabetes. Hence the maintenance or restoration of insulin sensitivity may prevent the onset of this disease. We previously showed that homozygous disruption of insulin receptor substrate-1 (IRS-1) in mice resulted in insulin resistance but not diabetes. Here, we have explored the mechanism of systemic insulin resistance in these mice and used adenovirus mediated gene therapy to restore their insulin sensitivity. Mice expressing the IRS-1transgene showed almost normal insulin sensitivity. Expression of an IRS-1 mutant (IRS-1Deltap85) lacking the binding site for the p85 subunit of phosphatidylinositol 3-kinase (PI3K) also restored insulin sensitivity, although PI3K is known to play a crucial role in insulin's metabolic responses. Protein kinase B (PKB) activity in liver was decreased in null mice compared with the wild-type and the null mice expressing IRS-1 or IRS-1Deltap85. In primary hepatocytes isolated from null mice, expression of IRS-1 enhanced both PI3K and PKB activities, but expression of IRS-1Deltap85 enhanced only PKB. These data suggest that PKB in liver plays a pivotal role in systemic glucose homeostasis and that PKB activation might be sufficient for reducing insulin resistance even without full activation of PI3K. PMID- 10811852 TI - A human class II MHC-derived peptide antagonizes phosphatidylinositol 3-kinase to block IL-2 signaling. AB - MHC molecules bind antigenic peptides and present them to T cells. There is a growing body of evidence that MHC molecules also serve other functions. We and others have described synthetic peptides derived from regions of MHC molecules that inhibit T-cell proliferation or cytotoxicity in an allele-nonspecific manner that is independent of interaction with the T-cell receptor. In this report, we describe the mechanism of action of a synthetic MHC class II-derived peptide that blocks T-cell activation induced by IL-2. Both this peptide, corresponding to residues 65-79 of DQA*03011 (DQ 65-79), and rapamycin inhibit p70 S6 kinase activity, but only DQ 65-79 blocks Akt kinase activity, placing the effects of DQ 65-79 upstream of mTOR, a PI kinase family member. DQ 65-79, but not rapamycin, inhibits phosphatidylinositol 3-kinase (PI 3-kinase) activity in vitro. The peptide is taken up by cells, as demonstrated by confocal microscopy. These findings indicate that DQ 65-79 acts as an antagonist with PI 3-kinase, repressing downstream signaling events and inhibiting proliferation. Understanding the mechanism of action of immunomodulatory peptides may provide new insights into T-cell activation and allow the development of novel immunosuppressive agents. PMID- 10811853 TI - Eosinophils generate brominating oxidants in allergen-induced asthma. AB - Eosinophils promote tissue injury and contribute to the pathogenesis of allergen triggered diseases like asthma, but the chemical basis of damage to eosinophil targets is unknown. We now demonstrate that eosinophil activation in vivo results in oxidative damage of proteins through bromination of tyrosine residues, a heretofore unrecognized pathway for covalent modification of biologic targets in human tissues. Mass spectrometric studies demonstrated that 3-bromotyrosine serves as a specific "molecular fingerprint" for proteins modified through the eosinophil peroxidase-H(2)O(2) system in the presence of plasma levels of halides. We applied a localized allergen challenge to model the effects of eosinophils and brominating oxidants in human lung injury. Endobronchial biopsy specimens from allergen-challenged lung segments of asthmatic, but not healthy control, subjects demonstrated significant enrichments in eosinophils and eosinophil peroxidase. Baseline levels of 3-bromotyrosine in bronchoalveolar lavage (BAL) proteins from mildly allergic asthmatic individuals were modestly but not statistically significantly elevated over those in control subjects. After exposure to segmental allergen challenge, lung segments of asthmatics, but not healthy control subjects, exhibited a >10-fold increase in BAL 3 bromotyrosine content, but only two- to threefold increases in 3-chlorotyrosine, a specific oxidation product formed by neutrophil- and monocyte-derived myeloperoxidase. These results identify reactive brominating species produced by eosinophils as a distinct class of oxidants formed in vivo. They also reveal eosinophil peroxidase as a potential therapeutic target for allergen-triggered inflammatory tissue injury in humans. PMID- 10811854 TI - Estrogen potentiates treatment with T-cell receptor protein of female mice with experimental encephalomyelitis. AB - Transgenic mice expressing the BV8S2 chain, which is specific for the myelin basic protein determinant Ac1-11, possess a naturally induced set of regulatory T cells directed against BV8S2. Further activation of anti-BV8S2 T cells in male mice with recombinant BV8S2 protein can inhibit IFN-gamma release by Ac1-11 specific T cells through a cytokine-driven mechanism and prevent induction of experimental autoimmune encephalomyelitis (EAE). In contrast, naive female mice possess fewer anti-BV8S2-reactive T cells, and treatment with BV8S2 delayed but did not prevent EAE. We here demonstrate that combining T-cell receptor (TCR) vaccination with supplemental estrus doses of estrogen potentiated IL-10 production by anti-BV8S2-reactive T cells and induced Ac1-11-specific T cells to produce IL-10 and TGF-beta. This combined treatment resulted in full protection against EAE, which was not observed with either therapy alone. These findings imply that supplemental estrogen can enhance the efficacy of TCR-based immunotherapy for autoimmune diseases that predominate in females. PMID- 10811856 TI - Surprising pleiotropy of nerve growth factor in the treatment of experimental autoimmune encephalomyelitis. PMID- 10811855 TI - Effect of regulated expression of human cyclooxygenase isoforms on eicosanoid and isoeicosanoid production in inflammation. AB - To examine the role of cyclooxygenase (COX) isozymes in prostaglandin formation and oxidant stress in inflammation, we administered to volunteer subjects placebo or bolus injections of lipopolysaccharide (LPS), which caused a dose-dependent increase in temperature, heart rate, and plasma cortisol. LPS caused also dose dependent elevations in urinary excretion of 2,3-dinor 6-keto PGF(1alpha) (PGI-M) and 11-dehydro thromboxane B(2) (Tx-M). Platelet COX-1 inhibition by chronic administration of low-dose aspirin before LPS did not alter the symptomatic and febrile responses to LPS, but the increment in urinary PGI-M and Tx-M were both partially depressed. Pretreatment with ibuprofen, a nonspecific COX inhibitor, attenuated the febrile and systemic response to LPS and inhibited prostanoid biosynthesis. Both celecoxib, a selective COX-2 inhibitor, and ibuprofen attenuated the pyrexial, but not the chronotropic, response to LPS. Experimental endotoxemia caused differential expression of the COX isozymes in monocytes and polymorphonuclear leucocytes ex vivo. LPS also increased urinary iPF(2alpha)-III, iPF(2alpha)-VI, and 8,12-iso-iPF(2alpha)-VI, isoprostane (iP) indices of lipid peroxidation, and none of the drugs blunted this response. These studies indicate that (a) although COX-2 predominates, both COX isozymes are induced and contribute to the prostaglandin response to LPS in humans; (b) COX activation contributes undetectably to lipid peroxidation induced by LPS; and (c) COX-2, but not COX-1, contributes to the constitutional response to LPS in humans. PMID- 10811857 TI - The old and the restless. PMID- 10811858 TI - Rearrangement of immunoglobulin genes in shark germ cells. AB - The variable (V), (diversity [D]), and joining (J) region recombinases (recombination activating genes [RAGs]) can perform like transposases and are thought to have initiated development of the adaptive immune system in early vertebrates by splitting archaic V genes with transposable elements. In cartilaginous fishes, the immunoglobulin (Ig) light chain genes are organized as multiple VJ-constant (C) clusters; some loci are capable of rearrangement while others contain fused VJ. The latter may be key to understanding the evolutionary role of RAG. Are they relics of the archaic genes, or are they results of rearrangement in germ cells? Our data suggest that some fused VJ genes are not only recently rearranged, but also resulted from RAG-like activity involving hairpin intermediates. Expression studies show that these, like some other germline-joined Ig sequences, are expressed at significant levels only early in ontogeny. We suggest that a rejoined Ig gene may not merely be a sequence restricting antibody diversity, but is potentially a novel receptor no longer tied to somatic RAG expression and rearrangement. From the combined data, we arrived at the unexpected conclusion that, in some vertebrates, RAG is still an active force in changing the genome. PMID- 10811859 TI - Human CD4(+) T lymphocytes consistently respond to the latent Epstein-Barr virus nuclear antigen EBNA1. AB - The Epstein-Barr virus (EBV)-encoded nuclear antigen EBNA1 is critical for the persistence of the viral episome in replicating EBV-transformed human B cells. Therefore, all EBV-induced tumors express this foreign antigen. However, EBNA1 is invisible to CD8(+) cytotoxic T lymphocytes because its Gly/Ala repeat domain prevents proteasome-dependent processing for presentation on major histocompatibility complex (MHC) class I. We now describe that CD4(+) T cells from healthy adults are primed to EBNA1. In fact, among latent EBV antigens that stimulate CD4(+) T cells, EBNA1 is preferentially recognized. We present evidence that the CD4(+) response may provide a protective role, including interferon gamma secretion and direct cytolysis after encounter of transformed B lymphocyte cell lines (B-LCLs). Dendritic cells (DCs) process EBNA1 from purified protein and from MHC class II-mismatched, EBNA1-expressing cells including B-LCLs. In contrast, B-LCLs and Burkitt's lymphoma lines likely present EBNA1 after endogenous processing, as their capacity to cross-present from exogenous sources is weak or undetectable. By limiting dilution, there is a tight correlation between the capacity of CD4(+) T cell lines to recognize autologous B-LCL expressing EBNA1 and DCs that have captured EBNA1. Therefore, CD4(+) T cells can respond to the EBNA1 protein that is crucial for EBV persistence. We suggest that this immune response is initiated in vivo by DCs that present EBV-infected B cells, and that EBNA1-specific CD4(+) T cell immunity be enhanced to prevent and treat EBV-associated malignancies. PMID- 10811860 TI - Dendritic cells discriminate between yeasts and hyphae of the fungus Candida albicans. Implications for initiation of T helper cell immunity in vitro and in vivo. AB - The fungus Candida albicans behaves as a commensal as well as a true pathogen of areas highly enriched in dendritic cells, such as skin and mucosal surfaces. The ability of the fungus to reversibly switch between unicellular yeast to filamentous forms is thought to be important for virulence. However, whether it is the yeast or the hyphal form that is responsible for pathogenicity is still a matter of debate. Here we show the interaction, and consequences, of different forms of C. albicans with dendritic cells. Immature myeloid dendritic cells rapidly and efficiently phagocytosed both yeasts and hyphae of the fungus. Phagocytosis occurred through different phagocytic morphologies and receptors, resulting in phagosome formation. However, hyphae escaped the phagosome and were found lying free in the cytoplasm of the cells. In vitro, ingestion of yeasts activated dendritic cells for interleukin (IL)-12 production and priming of T helper type 1 (Th1) cells, whereas ingestion of hyphae inhibited IL-12 and Th1 priming, and induced IL-4 production. In vivo, generation of antifungal protective immunity was induced upon injection of dendritic cells ex vivo pulsed with Candida yeasts but not hyphae. The immunization capacity of yeast-pulsed dendritic cells was lost in the absence of IL-12, whereas that of hypha-pulsed dendritic cells was gained in the absence of IL-4. These results indicate that dendritic cells fulfill the requirement of a cell uniquely capable of sensing the two forms of C. albicans in terms of type of immune responses elicited. By the discriminative production of IL-12 and IL-4 in response to the nonvirulent and virulent forms of the fungus, dendritic cells appear to meet the challenge of Th priming and education in C. albicans saprophytism and infections. PMID- 10811861 TI - Suppression of tumorigenicity in breast cancer cells by the microfilament protein profilin 1. AB - Differential display screening was used to reveal differential gene expression between the tumorigenic breast cancer cell line CAL51 and nontumorigenic microcell hybrids obtained after transfer of human chromosome 17 into CAL51. The human profilin 1 (PFN1) gene was found overexpressed in the microcell hybrid clones compared with the parental line, which displayed a low profilin 1 level. A comparison between several different tumorigenic breast cancer cell lines with nontumorigenic lines showed consistently lower profilin 1 levels in the tumor cells. Transfection of PFN1 cDNA into CAL51 cells raised the profilin 1 level, had a prominent effect on cell growth, cytoskeletal organization and spreading, and suppressed tumorigenicity of the stable, PFN1-overexpressing cell clones in nude mice. Immunohistochemical analysis revealed intermediate and low levels of profilin 1 in different human breast cancers. These results suggest profilin 1 as a suppressor of the tumorigenic phenotype of breast cancer cells. PMID- 10811862 TI - Incomplete CD8(+) T lymphocyte differentiation as a mechanism for subdominant cytotoxic T lymphocyte responses to a viral antigen. AB - CD8(+) cytotoxic T lymphocytes (CTLs) recognize antigen in the context of major histocompatibility complex (MHC) class I molecules. Class I epitopes have been classified as dominant or subdominant depending on the magnitude of the CTL response to the epitope. In this report, we have examined the in vitro memory CTL response of H-2(d) haplotype murine CD8(+) T lymphocytes specific for a dominant and subdominant epitope of influenza hemagglutinin using activation marker expression and staining with soluble tetrameric MHC-peptide complexes. Immune CD8(+) T lymphocytes specific for the dominant HA204-210 epitope give rise to CTL effectors that display activation markers, stain with the HA204 tetramer, and exhibit effector functions (i.e., cytolytic activity and cytokine synthesis). In contrast, stimulation of memory CD8(+) T lymphocytes directed to the subdominant HA210-219 epitope results in the generation of a large population of activated CD8(+) T cells that exhibit weak cytolytic activity and fail to stain with the HA210 tetramer. After additional rounds of restimulation with antigen, the HA210 219-specific subdominant CD8(+) T lymphocytes give rise to daughter cells that acquire antigen-specific CTL effector activity and transition from a HA210 tetramer-negative to a tetramer-positive phenotype. These results suggest a novel mechanism to account for weak CD8(+) CTL responses to subdominant epitopes at the level of CD8(+) T lymphocyte differentiation into effector CTL. The implications of these findings for CD8(+) T lymphocyte activation are discussed. PMID- 10811863 TI - Comparative analysis of genetically modified dendritic cells and tumor cells as therapeutic cancer vaccines. AB - We have directly compared the efficacy of two immunotherapeutic strategies for the treatment of cancer: "vaccination" of tumor-bearing mice with genetically modified dendritic cells (DCs), and vaccination with genetically modified tumor cells. Using several different preexisting tumor models that make use of B16F10 melanoma cells expressing a target tumor antigen (human melanoma-associated gene [MAGE]-1), we found that vaccination with bone marrow-derived DCs engineered to express MAGE-1 via adenoviral-mediated gene transfer led to a dramatic decrease in the number of metastases in a lung metastasis model, and led to prolonged survival and some long-term cures in a subcutaneous preexisting tumor model. In contrast, vaccination with granulocyte/macrophage colony-stimulating factor (GM CSF)-transduced tumor cells, previously shown to induce potent antitumor immunity in standard tumor challenge assays, led to a decreased therapeutic effect in the metastasis model and no effect in the subcutaneous tumor model. Further engineering of DCs to express either GM-CSF, tumor necrosis factor alpha, or CD40 ligand via retroviral-mediated gene transfer, led to a significantly increased therapeutic effect in the subcutaneous tumor model. The immunological mechanism, as shown for GM-CSF-transduced DCs, involves MAGE-1-specific CD4(+) and CD8(+) T cells. Expression of GM-CSF by DCs led to enhanced cytotoxic T lymphocyte activity, potentially mediated by increased numbers of DCs in draining lymph nodes. Our results suggest that clinical studies involving the vaccination with genetically modified DCs may be warranted. PMID- 10811864 TI - Apoptotic protease activating factor 1 (Apaf-1)-independent cell death suppression by Bcl-2. AB - Reportedly, antiapoptotic Bcl-2 family proteins suppress apoptosis by binding to and inhibiting members of the CED-4 family of caspase activators. To explore this question, we used embryonic stem (ES) cells in which one (-/+) or both (-/-) copies of the gene encoding apoptotic protease activating factor 1 (Apaf-1), a CED-4 homologue, were disrupted by homologous recombination. Stable clones of heterozygous (-/+) and homozygous (-/-) Apaf-1 knockout ES cells that overexpressed Bcl-2 were generated. Withdrawal of serum growth factors or stimulation of heterozygous ES cells with staurosporine (STS), ultraviolet (UV)B irradiation, etoposide (VP16), or cisplatin induced apoptosis followed by cell death (determined by failure to exclude propidium iodide dye). These cell death stimuli also induced activation of several types of caspases and loss of mitochondrial membrane potential (DeltaPsi) in heterozygous (+/-) Apaf-1 knockout ES cells. In addition, overexpression of Bcl-2 protected against these events in Apaf-1-expressing ES cells. In contrast, STS, UVB, and VP16 induced little or no caspase activation and apoptosis in homozygous (-/-) Apaf-1 knockout ES cells. Nevertheless, Apaf-1-deficient ES cells subjected to these cell death stimuli or deprived of growth factors did eventually die through a nonapoptotic mechanism associated with loss of DeltaPsi. Moreover, Bcl-2 overprotection preserved DeltaPsi, reduced the percentage of Apaf-1(-/)- ES cells undergoing cell death, and increased clonigenic survival. The extent of Bcl-2-mediated cytoprotection was not significantly different for heterozygous (-/+) versus homozygous (-/-) Apaf-1 knockout cells. Furthermore, although Bcl-2 could be readily coimmunoprecipitated with Bax, associations with Apaf-1 were undetectable under conditions where Apaf-1 interactions with procaspase-9 were observed. We conclude that Bcl-2 has cytoprotective functions independent of Apaf-1, preserving mitochondrial function through a caspase-independent mechanism. PMID- 10811865 TI - Protein kinase B regulates T lymphocyte survival, nuclear factor kappaB activation, and Bcl-X(L) levels in vivo. AB - The serine/threonine kinase protein kinase B (PKB)/Akt mediates cell survival in a variety of systems. We have generated transgenic mice expressing a constitutively active form of PKB (gag-PKB) to examine the effects of PKB activity on T lymphocyte survival. Thymocytes and mature T cells overexpressing gag-PKB displayed increased active PKB, enhanced viability in culture, and resistance to a variety of apoptotic stimuli. PKB activity prolonged the survival of CD4(+)CD8(+) double positive (DP) thymocytes in fetal thymic organ culture, but was unable to prevent antigen-induced clonal deletion of thymocytes expressing the major histocompatibility complex class I-restricted P14 T cell receptor (TCR). In mature T lymphocytes, PKB can be activated in response to TCR stimulation, and peptide-antigen-specific proliferation is enhanced in T cells expressing the gag-PKB transgene. Both thymocytes and T cells overexpressing gag PKB displayed elevated levels of the antiapoptotic molecule Bcl-X(L). In addition, the activation of peripheral T cells led to enhanced nuclear factor (NF)-kappaB activation via accelerated degradation of the NF-kappaB inhibitory protein IkappaBalpha. Our data highlight a physiological role for PKB in promoting survival of DP thymocytes and mature T cells, and provide evidence for the direct association of three major survival molecules (PKB, Bcl-X(L), and NF kappaB) in vivo in T lymphocytes. PMID- 10811866 TI - Bruton's tyrosine kinase links the B cell receptor to nuclear factor kappaB activation. AB - The recognition of antigen by membrane immunoglobulin M (mIgM) results in a complex series of signaling events in the cytoplasm leading to gene activation. Bruton's tyrosine kinase (BTK), a member of the Tec family of tyrosine kinases, is essential for the full repertoire of IgM signals to be transduced. We examined the ability of BTK to regulate the nuclear factor (NF)-kappaB/Rel family of transcription factors, as the activation of these factors is required for a B cell response to mIgM. We found greatly diminished IgM- but not CD40-mediated NF kappaB/Rel nuclear translocation and DNA binding in B cells from X-linked immunodeficient (xid) mice that harbor an R28C mutation in btk, a mutation that produces a functionally inactive kinase. The defect was due, in part, to a failure to fully degrade the inhibitory protein of NF-kappaB, IkappaBalpha. Using a BTK-deficient variant of DT40 chicken B cells, we found that expression of wild type or gain-of-function mutant BTK, but not the R28C mutant, reconstituted NF kappaB activity. Thus, BTK is essential for activation of NF-kappaB via the B cell receptor. PMID- 10811867 TI - Bruton's tyrosine kinase is required for activation of IkappaB kinase and nuclear factor kappaB in response to B cell receptor engagement. AB - Mutations in the gene encoding Bruton's tyrosine kinase (btk) cause the B cell deficiency diseases X-linked agammaglobulinemia (XLA) in humans and X-linked immunodeficiency (xid) in mice. In vivo and in vitro studies indicate that the BTK protein is essential for B cell survival, cell cycle progression, and proliferation in response to B cell antigen receptor (BCR) stimulation. BCR stimulation leads to the activation of transcription factor nuclear factor (NF) kappaB, which in turn regulates genes controlling B cell growth. We now demonstrate that a null mutation in btk known to cause the xid phenotype prevents BCR-induced activation of NF-kappaB. This defect can be rescued by reconstitution with wild-type BTK. This mutation also interferes with BCR-directed activation of IkappaB kinase (IKK), which normally targets the NF-kappaB inhibitor IkappaBalpha for degradation. Taken together, these findings indicate that BTK couples IKK and NF-kappaB to the BCR. Interference with this coupling mechanism may contribute to the B cell deficiencies observed in XLA and xid. PMID- 10811868 TI - A key role for CC chemokine receptor 4 in lipopolysaccharide-induced endotoxic shock. AB - CC chemokine receptor (CCR)4, a high affinity receptor for the CC chemokines thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), is expressed in the thymus and spleen, and also by peripheral blood T cells, macrophages, platelets, and basophils. Recent studies have shown that CCR4 is the major chemokine receptor expressed by T helper type 2 (Th2) polarized cells. To study the in vivo role of CCR4, we have generated CCR4-deficient (CCR4( /-)) mice by gene targeting. CCR4(-/-) mice developed normally. Splenocytes and thymocytes isolated from the CCR4(-/-) mice failed to respond to the CCR4 ligands TARC and MDC, as expected, but also surprisingly did not undergo chemotaxis in vitro in response to macrophage inflammatory protein (MIP)-1alpha. The CCR4 deletion had no effect on Th2 differentiation in vitro or in a Th2-dependent model of allergic airway inflammation. However, CCR4(-/-) mice exhibited significantly decreased mortality on administration of high or low dose bacterial lipopolysaccharide (LPS) compared with CCR4(+/+) mice. After high dose LPS treatment, serum levels of tumor necrosis factor alpha, interleukin 1beta, and MIP-1alpha were reduced in CCR4(-/-) mice, and decreased expression of MDC and MIP-2 mRNA was detected in peritoneal exudate cells. Analysis of peritoneal lavage cells from CCR4(-/)- mice by flow cytometry also revealed a significant decrease in the F4/80(+) cell population. This may reflect a defect in the ability of the CCR4(-/-) macrophages to be retained in the peritoneal cavity. Taken together, our data reveal an unexpected role for CCR4 in the inflammatory response leading to LPS-induced lethality. PMID- 10811869 TI - The streptococcal superantigen SMEZ exhibits wide allelic variation, mosaic structure, and significant antigenic variation. AB - The frequencies of the newly identified streptococcal superantigen genes smez, spe-g, and spe-h were determined in a panel of 103 clinical isolates collected between 1976 and 1998 at various locations throughout New Zealand. smez and spe-g were found in every group A Streptococcus (GAS) isolate, suggesting a chromosomal location. The spe-h gene was found in only 24% of the GAS isolates and is probably located on a mobile DNA element. The smez gene displays extensive allelic variation and appears to be in linkage equilibrium with the M/emm type. 22 novel smez alleles were identified from 21 different M/emm types in addition to the already reported alleles smez and smez-2 with sequence identities between 94. 5 and 99.9%. Three alleles are nonfunctional due to a single base pair deletion. The remaining 21 alleles encode distinct SMEZ variants. The mosaic structure of the smez gene suggests that this polymorphism has arisen from homologous recombination events rather than random point mutation. The recently resolved SMEZ-2 crystal structure shows that the polymorphic residues are mainly surface exposed and scattered over the entire protein. The allelic variation did not affect either Vbeta specificity or potency, but did result in significant antigenic differences. Neutralizing antibody responses of individual human sera against different SMEZ variants varied significantly. 98% of sera completely neutralized SMEZ-1, but only 85% neutralized SMEZ-2, a very potent variant that has not yet been found in any New Zealand isolate. SMEZ-specific Vbeta8 activity was found in culture supernatants of 66% of the GAS isolates, indicating a potential base for the development of a SMEZ targeting vaccine. PMID- 10811870 TI - Type I interferon as a powerful adjuvant for monocyte-derived dendritic cell development and activity in vitro and in Hu-PBL-SCID mice. AB - Type I interferons (IFNs) are cytokines exhibiting antiviral and antitumor effects, including multiple activities on immune cells. However, the importance of these cytokines in the early events leading to the generation of an immune response is still unclear. Here, we have investigated the effects of type I IFNs on freshly isolated granulocyte/macrophage colony-stimulating factor (GM-CSF) treated human monocytes in terms of dendritic cell (DC) differentiation and activity in vitro and in severe combined immunodeficiency mice reconstituted with human peripheral blood leukocytes (hu-PBL-SCID) mice. Type I IFNs induced a surprisingly rapid maturation of monocytes into short-lived tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-expressing DCs endowed with potent functional activities, superior with respect to the interleukin (IL)-4/GM CSF treatment, as shown by FACS((R)) analyses, mixed leukocyte reaction assays with allogeneic PBLs, and lymphocyte proliferation responses to HIV-1-pulsed autologous DCs. Type I IFN induced IL-15 production and strongly promoted a T helper cell type 1 response. Notably, injection of IFN-treated HIV-1-pulsed DCs in SCID mice reconstituted with autologous PBLs resulted in the generation of a potent primary immune response, as evaluated by the detection of human antibodies to various HIV-1 antigens. These results provide a rationale for using type I IFNs as vaccine adjuvants and support the concept that a natural alliance between these cytokines and monocytes/DCs represents an important early mechanism for connecting innate and adaptive immunity. PMID- 10811871 TI - Thrombospondin-1 is downregulated by anoxia and suppresses tumorigenicity of human glioblastoma cells. AB - Angiogenesis, the sprouting of new capillaries from preexisting blood vessels, results from a disruption of the balance between stimulatory and inhibitory factors. Here, we show that anoxia reduces expression of thrombospondin-1 (TSP 1), a natural inhibitor of angiogenesis, in glioblastoma cells. This suggests that reduced oxygen tension can promote angiogenesis not only by stimulating the production of inducers, such as vascular endothelial growth factor, but also by reducing the production of inhibitors. This downregulation may significantly contribute to glioblastoma development, since we show that an increase in TSP-1 expression is sufficient to strongly suppress glioblastoma cell tumorigenicity in vivo. PMID- 10811872 TI - Human nerve growth factor protects common marmosets against autoimmune encephalomyelitis by switching the balance of T helper cell type 1 and 2 cytokines within the central nervous system. AB - Multiple sclerosis is a demyelinating disorder of the central nervous system (CNS), in which an immune attack directed against myelin constituents causes myelin destruction and death of oligodendrocytes, the myelin-producing cells. Here, the efficacy of nerve growth factor (NGF), a growth factor for neurons and oligodendrocytes, in promoting myelin repair was evaluated using the demyelinating model of experimental allergic encephalomyelitis (EAE) in the common marmoset. Surprisingly, we found that NGF delayed the onset of clinical EAE and, pathologically, prevented the full development of EAE lesions. We demonstrate by immunocytochemistry that NGF exerts its antiinflammatory effect by downregulating the production of interferon gamma by T cells infiltrating the CNS, and upregulating the production of interleukin 10 by glial cells in both inflammatory lesions of EAE and normal-appearing CNS white matter. Thus, NGF, currently under investigation in human clinical trials as a neuronal trophic factor, may be an attractive candidate for therapy of autoimmune demyelinating disorders. PMID- 10811874 TI - Fast methods for the Eikonal and related Hamilton- Jacobi equations on unstructured meshes. AB - The Fast Marching Method is a numerical algorithm for solving the Eikonal equation on a rectangular orthogonal mesh in O(M log M) steps, where M is the total number of grid points. The scheme relies on an upwind finite difference approximation to the gradient and a resulting causality relationship that lends itself to a Dijkstra-like programming approach. In this paper, we discuss several extensions to this technique, including higher order versions on unstructured meshes in Rn and on manifolds and connections to more general static Hamilton Jacobi equations. PMID- 10811873 TI - Innate recognition of bacteria in human milk is mediated by a milk-derived highly expressed pattern recognition receptor, soluble CD14. AB - Little is known about innate immunity to bacteria after birth in the hitherto sterile fetal intestine. Breast-feeding has long been associated with a lower incidence of gastrointestinal infections and inflammatory and allergic diseases. We found in human breast milk a 48-kD polypeptide, which we confirmed by mass spectrometry and sequencing to be a soluble form of the bacterial pattern recognition receptor CD14 (sCD14). Milk sCD14 (m-sCD14) concentrations were up to 20-fold higher than serum sCD14 from nonpregnant, pregnant, or lactating women. In contrast, lipopolysaccharide (LPS)-binding protein was at very low levels. Mammary epithelial cells produced 48-kD sCD14. m-sCD14 mediated activation by LPS and whole bacteria of CD14 negative cells, including intestinal epithelial cells, resulting in release of innate immune response molecules. m-sCD14 was undetectable in the infant formulas and commercial (cows') milk tested, although it was present in bovine colostrum. These findings indicate a sentinel role for sCD14 in human milk during bacterial colonization of the gut, and suggest that m sCD14 may be involved in modulating local innate and adaptive immune responses, thus controlling homeostasis in the neonatal intestine. PMID- 10811875 TI - Ski acts as a co-repressor with Smad2 and Smad3 to regulate the response to type beta transforming growth factor. AB - The c-ski protooncogene encodes a transcription factor that binds DNA only in association with other proteins. To identify co-binding proteins, we performed a yeast two-hybrid screen. The results of the screen and subsequent co immunoprecipitation studies identified Smad2 and Smad3, two transcriptional activators that mediate the type beta transforming growth factor (TGF-beta) response, as Ski-interacting proteins. In Ski-transformed cells, all of the Ski protein was found in Smad3-containing complexes that accumulated in the nucleus in the absence of added TGF-beta. DNA binding assays showed that Ski, Smad2, Smad3, and Smad4 form a complex with the Smad/Ski binding element GTCTAGAC (SBE). Ski repressed TGF-beta-induced expression of 3TP-Lux, the natural plasminogen activator inhibitor 1 promoter and of reporter genes driven by the SBE and the related CAGA element. In addition, Ski repressed a TGF-beta-inducible promoter containing AP-1 (TRE) elements activated by a combination of Smads, Fos, and/or Jun proteins. Ski also repressed synergistic activation of promoters by combinations of Smad proteins but failed to repress in the absence of Smad4. Thus, Ski acts in opposition to TGF-beta-induced transcriptional activation by functioning as a Smad-dependent co-repressor. The biological relevance of this transcriptional repression was established by showing that overexpression of Ski abolished TGF-beta-mediated growth inhibition in a prostate-derived epithelial cell line. PMID- 10811876 TI - High throughput protein fold identification by using experimental constraints derived from intramolecular cross-links and mass spectrometry. AB - We have used intramolecular cross-linking, MS, and sequence threading to rapidly identify the fold of a model protein, bovine basic fibroblast growth factor (FGF) 2. Its tertiary structure was probed with a lysine-specific cross-linking agent, bis(sulfosuccinimidyl) suberate (BS(3)). Sites of cross-linking were determined by tryptic peptide mapping by using time-of-flight MS. Eighteen unique intramolecular lysine (Lys-Lys) cross-links were identified. The assignments for eight cross-linked peptides were confirmed by using post source decay MS. The interatomic distance constraints were all consistent with the tertiary structure of FGF-2. These relatively few constraints, in conjunction with threading, correctly identified FGF-2 as a member of the beta-trefoil fold family. To further demonstrate utility, we used the top-scoring homolog, IL-1beta, to build an FGF-2 homology model with a backbone error of 4.8 A (rms deviation). This method is fast, is general, uses small amounts of material, and is amenable to automation. PMID- 10811877 TI - Neuropeptide Y: A hypothalamic brake restraining the onset of puberty in primates. AB - The adult reproductive axis is driven by an intermittent discharge of gonadotropin-releasing hormone (GnRH) generated by a network of hypothalamic neurons known as the GnRH pulse generator. Although this signal generator is operational in infant primates, puberty in these species is delayed by activation shortly after birth of a central neural mechanism that holds GnRH release in check during juvenile development. Here, we show that, in the male rhesus monkey, the postnatal pattern in GnRH pulse generator activity is inversely related to that in neuropeptide Y (NPY) gene and protein expression in the mediobasal hypothalamus and that central administration of an NPY Y(1) receptor antagonist to juvenile animals elicits precocious GnRH release. Cell imaging indicated that the developmentally regulated NPY neurons may be located in regions dorsal to the arcuate nucleus. These findings lead us to propose that NPY is a fundamental component of the neurobiological brake restraining the onset of puberty in primates. PMID- 10811878 TI - The chemokine growth-related gene product beta protects rat cerebellar granule cells from apoptotic cell death through alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate receptors. AB - Cultured cerebellar granule neurons are widely used as a cellular model to study mechanisms of neuronal cell death because they undergo programmed cell death when switched from a culture medium containing 25 mM to one containing 5 mM K(+). We have found that the growth-related gene product beta (GRObeta) partially prevents the K(+)-depletion-induced cell death, and that the neuroprotective action of GRObeta on granule cells is mediated through the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) type of ionotropic glutamate receptors. GRObeta induced survival was suppressed by 6-cyano-7-nitroquinoxaline-2,3-dione, which is a specific antagonist of AMPA/kainate receptors; it was not affected by the inhibitor of N-methyl-D-aspartate receptors, 2-amino-5-phosphonopentanoic acid, and was comparable to the survival of granule cells induced by AMPA (10 microM) treatment. Moreover, GRObeta-induced neuroprotection was abolished when granule cells were treated with antisense oligonucleotides specific for the AMPA receptor subunits, which significantly reduced receptor expression, as verified by Western blot analysis with subunit-specific antibodies and by granule cell electrophysiological sensitivity to AMPA. Our data demonstrate that GRObeta is neurotrophic for cerebellar granule cells, and that this activity depends on AMPA receptors. PMID- 10811879 TI - Cerebral metabolic and cognitive decline in persons at genetic risk for Alzheimer's disease. AB - The major known genetic risk for Alzheimer's disease (AD), apolipoprotein E-4 (APOE-4), is associated with lowered parietal, temporal, and posterior cingulate cerebral glucose metabolism in patients with a clinical diagnosis of AD. To determine cognitive and metabolic decline patterns according to genetic risk, we investigated cerebral metabolic rates by using positron emission tomography in middle-aged and older nondemented persons with normal memory performance. A single copy of the APOE-4 allele was associated with lowered inferior parietal, lateral temporal, and posterior cingulate metabolism, which predicted cognitive decline after 2 years of longitudinal follow-up. For the 20 nondemented subjects followed longitudinally, memory performance scores did not decline significantly, but cortical metabolic rates did. In APOE-4 carriers, a 4% left posterior cingulate metabolic decline was observed, and inferior parietal and lateral temporal regions demonstrated the greatest magnitude (5%) of metabolic decline after 2 years. These results indicate that the combination of cerebral metabolic rates and genetic risk factors provides a means for preclinical AD detection that will assist in response monitoring during experimental treatments. PMID- 10811880 TI - Preventing ventricular fibrillation by flattening cardiac restitution. AB - Ventricular fibrillation is the leading cause of sudden cardiac death. In fibrillation, fragmented electrical waves meander erratically through the heart muscle, creating disordered and ineffective contraction. Theoretical and computer studies, as well as recent experimental evidence, have suggested that fibrillation is created and sustained by the property of restitution of the cardiac action potential duration (that is, its dependence on the previous diastolic interval). The restitution hypothesis states that steeply sloped restitution curves create unstable wave propagation that results in wave break, the event that is necessary for fibrillation. Here we present experimental evidence supporting this idea. In particular, we identify the action of the drug bretylium as a prototype for the future development of effective restitution based antifibrillatory agents. We show that bretylium acts in accord with the restitution hypothesis: by flattening restitution curves, it prevents wave break and thus prevents fibrillation. It even converts existing fibrillation, either to a periodic state (ventricular tachycardia, which is much more easily controlled) or to quiescent healthy tissue. PMID- 10811881 TI - The tetranucleotide UCAY directs the specific recognition of RNA by the Nova K homology 3 domain. AB - The Nova family of proteins are target antigens in the autoimmune disorder paraneoplastic opsoclonus-myoclonus ataxia and contain K-homology (KH)-type RNA binding domains. The Nova-1 protein has recently been shown to regulate alternative splicing of the alpha2 glycine receptor subunit pre-mRNA by binding to an intronic element containing repeats of the tetranucleotide UCAU. Here, we have used selection-amplification to demonstrate that the KH3 domain of Nova recognizes a single UCAY element in the context of a 20-base hairpin RNA; the UCAY tetranucleotide is optimally presented as a loop element of the hairpin scaffold and requires protein residues C-terminal to the previously defined KH domain. These results suggest that KH domains in general recognize tetranucleotide motifs and that biological RNA targets of KH domains may use either RNA secondary structure or repeated sequence elements to achieve high affinity and specificity of protein binding. PMID- 10811882 TI - Pseudoxanthoma elasticum: mutations in the MRP6 gene encoding a transmembrane ATP binding cassette (ABC) transporter. AB - Pseudoxanthoma elasticum (PXE), the prototypic heritable connective tissue disorder affecting the elastic structures in the body, manifests with cutaneous, ophthalmologic, and cardiovascular findings, with considerable morbidity and mortality. The molecular basis of PXE has remained unknown, but the disease locus has recently been mapped to an approximately 500-kb interval on chromosome 16p13.1, without evidence for locus heterogeneity. In this study, we report pathogenetic mutations in MRP6, a member of the ABC transporter gene family, in eight kindreds with PXE. The mutation detection strategy consisted of heteroduplex scanning of coding sequences in the MRP6 gene, which were amplified by PCR by using genomic DNA as template, followed by direct nucleotide sequencing. A total of 13 mutant MRP6 alleles were disclosed in the eight probands with PXE. These genetic lesions consisted of either single base pair substitutions resulting in missense, nonsense, or splice site mutations, or large deletions resulting in allelic loss of the MRP6 locus. Examination of clinically unaffected family members in four multiplex families identified heterozygous carriers, consistent with an autosomal recessive inheritance pattern. Collectively, identification of mutations in the MRP6 gene provides the basis to examine the pathomechanisms of PXE and allows development of DNA-based carrier detection, prenatal testing, and preimplantation genetic diagnosis in families with a history of this disease. PMID- 10811883 TI - Separation of presenilin function in amyloid beta-peptide generation and endoproteolysis of Notch. AB - Most of the genetically inherited Alzheimer's disease cases are caused by mutations in the presenilin genes, PS1 and PS2. PS mutations result in the enhanced production of the highly amyloidogenic 42/43 amino acid variant of amyloid beta-peptide (Abeta). We have introduced arbitrary mutations at position 286 of PS1, where a naturally occurring PS1 mutation has been described (L286V). Introduction of charged amino acids (L286E or L286R) resulted in an increase of Abeta42/43 production, which reached almost twice the level of the naturally occurring PS1 mutation. Although pathological Abeta production was increased, endoproteolysis of Notch and nuclear transport of its cytoplasmic domain was significantly inhibited. These results demonstrate that the biological function of PS proteins in the endoproteolysis of beta-amyloid precursor protein and Notch can be separated. PMID- 10811885 TI - Construction of hybrid peptide synthetases by module and domain fusions. AB - Nonribosomal peptide synthetases are modular enzymes that assemble peptides of diverse structures and important biological activities. Their modular organization provides a great potential for the rational design of novel compounds by recombination of the biosynthetic genes. Here we describe the extension of a dimodular system to trimodular ones based on whole-module fusion. The recombinant hybrid enzymes were purified to monitor product assembly in vitro. We started from the first two modules of tyrocidine synthetase, which catalyze the formation of the dipeptide dPhe-Pro, to construct such hybrid systems. Fusion of the second, proline-specific module with the ninth and tenth modules of the tyrocidine synthetases, specific for ornithine and leucine, respectively, resulted in dimodular hybrid enzymes exhibiting the combined substrate specificities. The thioesterase domain was fused to the terminal module. Upon incubation of these dimodular enzymes with the first tyrocidine module, TycA, incorporating dPhe, the predicted tripeptides dPhe-Pro-Orn and dPhe Pro-Leu were obtained at rates of 0.15 min(-1) and 2.1 min(-1). The internal thioesterase domain was necessary and sufficient to release the products from the hybrid enzymes and thereby facilitate a catalytic turnover. Our approach of whole module fusion is based on an improved definition of the fusion sites and overcomes the recently discovered editing function of the intrinsic condensation domains. The stepwise construction of hybrid peptide synthetases from catalytic subunits reinforces the inherent potential for the synthesis of novel, designed peptides. PMID- 10811884 TI - A multipotential beta -1,6-N-acetylglucosaminyl-transferase is encoded by bovine herpesvirus type 4. AB - The beta-1,6-N-acetylglucosaminyltransferase (beta1,6GnT) gene family encodes enzymes playing crucial roles in glycan synthesis. Important changes in beta1,6GnT expression are observed during development, oncogenesis, and immunodeficiency. The most characterized beta1,6GnTs in this gene family are the human (h) C2GnT-L and h-IGnT, which have core 2 [Galbeta1-->3(GlcNAcbeta1- >6)GalNAc] and I branching [GlcNAcbeta1-->3(GlcNAcbeta1-->6)Gal] activities, respectively. Recently, h-C2GnT-M was shown to be unique in forming core 2, core 4 [GlcNAcbeta1-->3(GlcNAcbeta1-->6)GalNAc], and I structures. To date, the beta1,6GnT gene family has been characterized only in mammals. Here, we describe that bovine herpesvirus type 4 (BHV-4) encodes a beta1,6GnT expressed during viral replication and exhibiting all of the core 2, core 4, and I branching activities. Sequencing of the BHV-4 genome revealed an ORF, hereafter called BORFF3-4, encoding a protein (pBORFF3-4) exhibiting 81.1%, 50.7%, and 36.6% amino acid identity with h-C2GnT-M, h-C2GnT-L, and h-IGnT, respectively. Reverse transcriptase-PCR analysis revealed that BORFF3-4 is expressed during BHV-4 replication. Expression of BORFF3-4 in Chinese hamster ovary cells directed the expression of core 2 branched oligosaccharides and I antigenic structures on the cell surface. Moreover, a soluble form of pBORFF3-4 had core 4 branching activity in addition to core 2 and I branching activities. Finally, infection of a C2GnT negative cell line with BHV-4 induced expression of core 2 branched oligosaccharides. This study extends the beta1,6GnT gene family to a viral gene and provides a model to study the biological functions of a beta1,6GnT in the context of viral infection. PMID- 10811886 TI - The multiple-race population of the United States: issues and estimates. AB - This paper presents national estimates of the population likely to identify with more than one race in the 2000 census as a result of a new federal policy allowing multiple racial identification. A large number of race-based public policies-including affirmative action and the redistricting provisions of the Voting Rights Act-may be affected by the shift of some 8-18 million people out of traditional single-race statistical groups. The declines in single-race populations resulting from the new classification procedure are likely to be greater in magnitude than the net undercount in the U.S. census at the center of the controversy over using census sampling. Based on ancestry data in the 1990 census and experimental survey results from the 1995 Current Population Survey, we estimate that 3. 1-6.6% of the U.S. population is likely to mark multiple races. Our results are substantially higher than those suggested by previous research and have implications for the coding, reporting, and use of multiple response racial data by government and researchers. The change in racial classification may pose new conundrums for the implementation of race-based public policies, which have faced increasing criticism in recent years. PMID- 10811887 TI - A syntactic specialization for Broca's area. AB - Despite numerous aphasia and functional imaging studies, the exact correlation between cortical language areas and subcomponents of the linguistic system has not been established. Here, we used functional MRI to identify cortical areas specifically involved in syntactic processing. An experimental design contrasted sentences containing grammatical errors with sentences containing spelling errors. The ungrammatical sentences produced more activation in cortical language areas than did the sentences with spelling errors, and the difference in activation was significantly greater in Broca's area than in Wernicke's area or in the angular gyrus/supramarginal gyrus. The present findings provide direct evidence of a syntactic specialization for Broca's area and establish the existence of distinct modules for our knowledge of language. PMID- 10811888 TI - UvrA and UvrB suppress illegitimate recombination: synergistic action with RecQ helicase. AB - Illegitimate recombination is a major cause of genetic instability in prokaryotes as well as in eukaryotes. This recombination usually occurs at a low frequency, but it is greatly enhanced by UV irradiation or other environmental stresses. DNA damages produced by these environmental stresses are thought to induce DNA double strand breaks, leading to illegitimate recombination. In this paper we show that UV-induced illegitimate recombination is enhanced by mutations of nucleotide excision repair genes, uvrA or uvrB, and partially by uvrC mutation, but not by uvrD mutation. Unexpectedly, the recombination was enhanced by the uvrA uvrB double mutation even without UV irradiation, but the uvrB uvrC double mutation has not shown this effect, suggesting that illegitimate recombination is mostly suppressed by UvrA and UvrB. Moreover, illegitimate recombination was synergistically enhanced by the recQ uvrA double mutation. In addition, overproduction of the UvrA protein suppressed the hyperrecombination phenotype of the recQ or uvrB mutant, but it did not affect the UV-sensitive phenotype of the uvrB mutant. We concluded that the UvrAB complex suppresses illegitimate recombination in a pathway shared with RecQ helicase. In addition, UvrA protein alone can suppress illegitimate recombination in the pathway, in which RecQ helicase and UvrAB complex work. Possible functions of the proteins involved in these pathways are also discussed. PMID- 10811889 TI - Effective inhibition of human cytomegalovirus gene expression and replication by a ribozyme derived from the catalytic RNA subunit of RNase P from Escherichia coli. AB - A sequence-specific ribozyme (M1GS RNA) derived from the catalytic RNA subunit of RNase P from Escherichia coli was used to target the overlapping exon 3 region of the mRNAs encoding the major transcription regulatory proteins IE1 and IE2 of human cytomegalovirus. A reduction of more than 80% in the expression levels of IE1 and IE2 and a reduction of about 150-fold in viral growth were observed in human cells that stably expressed the ribozyme. In contrast, a reduction of less than 10% in the IE1/IE2 expression and viral growth was observed in cells that either did not express the ribozyme or produced a "disabled" ribozyme that carried mutations that abolished its catalytic activity. Examination of the expression of several other viral early and late genes in the cells that expressed the M1GS ribozyme further revealed an overall reduction of at least 80% in their expression. These results are consistent with the notion that the antiviral effects in these cells are due to the fact that the ribozyme specifically inhibits the expression of IE1 and IE2 and, consequently, abolishes the expression of viral early and late genes as well as viral growth. Our study is the first, to our knowledge, to use M1GS ribozyme for inhibiting human cytomegalovirus replication and demonstrates the utility of this ribozyme for antiviral applications. PMID- 10811890 TI - Polyglutamine aggregates alter protein folding homeostasis in Caenorhabditis elegans. AB - Expansion of polyglutamine repeats in several unrelated proteins causes neurodegenerative diseases with distinct but related pathologies. To provide a model system for investigating common pathogenic features, we have examined the behavior of polyglutamine expansions expressed in Caenorhabditis elegans. The expression of polyglutamine repeats as green fluorescent protein (GFP)-fusion proteins in body wall muscle cells causes discrete cytoplasmic aggregates that appear early in embryogenesis and correlates with a delay in larval to adult development. The heat shock response is activated idiosyncratically in individual cells in a polyglutamine length-dependent fashion. The toxic effect of polyglutamine expression and the formation of aggregates can be reversed by coexpression of the yeast chaperone Hsp104. The altered homeostasis associated with polyglutamine aggregates causes both the sequestration of an otherwise soluble protein with shorter arrays of glutamine repeats and the relocalization of a nuclear glutamine-rich protein. These observations of induced aggregation and relocalization have implications for disorders involving protein aggregation. PMID- 10811892 TI - Reduced genetic variation and the success of an invasive species. AB - Despite the severe ecological and economic damage caused by introduced species, factors that allow invaders to become successful often remain elusive. Of invasive taxa, ants are among the most widespread and harmful. Highly invasive ants are often unicolonial, forming supercolonies in which workers and queens mix freely among physically separate nests. By reducing costs associated with territoriality, unicolonial species can attain high worker densities, allowing them to achieve interspecific dominance. Here we examine the behavior and population genetics of the invasive Argentine ant (Linepithema humile) in its native and introduced ranges, and we provide a mechanism to explain its success as an invader. Using microsatellite markers, we show that a population bottleneck has reduced the genetic diversity of introduced populations. This loss is associated with reduced intraspecific aggression among spatially separate nests, and leads to the formation of interspecifically dominant supercolonies. In contrast, native populations are more genetically variable and exhibit pronounced intraspecific aggression. Although reductions in genetic diversity are generally considered detrimental, these findings provide an example of how a genetic bottleneck can lead to widespread ecological success. In addition, these results provide insights into the origin and evolution of unicoloniality, which is often considered a challenge to kin selection theory. PMID- 10811891 TI - Steroid hormones induce HMG1 overexpression and sensitize breast cancer cells to cisplatin and carboplatin. AB - Cisplatin is an anticancer drug that has enjoyed remarkable success against testicular tumors, but dose limiting side-effects have limited its application against a broader range of cancers. Previous studies have shown that high mobility group (HMG) domain proteins such as HMG1 sensitize cells to cisplatin by shielding its major DNA adducts from nucleotide excision repair. Estrogen treatment increases HMG1 mRNA levels in breast cancer MCF-7 cells. Herein, we describe that treatment of human cancer cells having steroid hormone receptors with the appropriate hormone, estrogen and/or progesterone, significantly increases the potency of cisplatin and its analogue carboplatin by causing the overexpression of HMG1. These findings suggest that the proper combination of these drugs, which are already approved by the Food and Drug Administration, could have potential benefit in treating tumors such as ovarian or breast that carry the hormone receptors. PMID- 10811893 TI - The yeast A kinases differentially regulate iron uptake and respiratory function. AB - Yeast has three A kinase catalytic subunits, which have greater than 75% identity and are encoded by the TPK genes (TPK1, TPK2, and TPK3) [Toda, T., Cameron, S., Sass, P., Zoller, M. & Wigler, M. (1987) Cell 50, 277-287]. Although they are redundant for viability, the three A kinases are not redundant for pseudohyphal growth [Robertson, L. S. & Fink, G. R. (1998) Proc. Natl. Acad. Sci. USA 95, 13783-13787; Pan, X. & Heitman, J. (1999) Mol. Cell. Biol. 19, 4874-4887]; Tpk2, but not Tpk1 or Tpk3, is required for pseudohyphal growth. Genome-wide transcriptional profiling has revealed unique signatures for each of the three A kinases leading to the identification of additional functional diversity among these proteins. Tpk2 negatively regulates genes involved in iron uptake and positively regulates genes involved in trehalose degradation and water homeostasis. Tpk1 is required for the derepression of branched chain amino acid biosynthesis genes that seem to have a second role in the maintenance of iron levels and DNA stability within mitochondria. The fact that TPK2 mutants grow better than wild types on nonfermentable carbon sources and on media deficient in iron supports the unique role of Tpk2 in respiratory growth and carbon source use. PMID- 10811895 TI - What angle-resolved photoemission experiments tell about the microscopic theory for high-temperature superconductors. AB - Recent angular-resolved photoemission experiments on high-temperature superconductors are consistent with a phenomenological description of the normal state of these materials as marginal Fermi liquids. The experiments also provide constraints on microscopic theories. PMID- 10811894 TI - Domain organization and flavin adenine dinucleotide-binding determinants in the aerotaxis signal transducer Aer of Escherichia coli. AB - Aerotactic responses in Escherichia coli are mediated by the membrane transducer Aer, a recently identified member of the superfamily of PAS domain proteins, which includes sensors of light, oxygen, and redox state. Initial studies of Aer suggested that it might use a flavin adenine dinucleotide (FAD) prosthetic group to monitor cellular redox changes. To test this idea, we purified lauryl maltoside-solubilized Aer protein by His-tag affinity chromatography and showed by high performance liquid chromatography, mass spectrometry, and absorbance spectroscopy that it bound FAD noncovalently. Polypeptide fragments spanning the N-terminal 290 residues of Aer, which contains the PAS motif, were able to bind FAD. Fusion of this portion of Aer to the flagellar signaling domain of Tsr, the serine chemoreceptor, yielded a functional aerotaxis transducer, demonstrating that the FAD-binding portion of Aer is sufficient for aerosensing. Aerotaxis defective missense mutants identified two regions, in addition to the PAS domain, that play roles in FAD binding. Those regions flank a central hydrophobic segment needed to anchor Aer to the cytoplasmic membrane. They might contact the FAD ligand directly or stabilize the FAD-binding pocket. However, their lack of sequence conservation in Aer homologs of other bacteria suggests that they play less direct roles in FAD binding. One or both regions probably also play important roles in transmitting stimulus-induced conformational changes to the C terminal flagellar signaling domain to trigger aerotactic behavioral responses. PMID- 10811896 TI - IL-10 is necessary for the expression of airway hyperresponsiveness but not pulmonary inflammation after allergic sensitization. AB - Cytokines play an important role in modulating inflammatory responses and, as a result, airway tone. IL-10 is a regulatory cytokine that has been suggested for treatment of asthma because of its immunosuppressive and anti-inflammatory properties. In contrast to these suggestions, we demonstrate in a model of allergic sensitization that mice deficient in IL-10 (IL-10-/-) develop a pulmonary inflammatory response but fail to exhibit airway hyperresponsiveness in both in vitro and in vivo assessments of lung function. Reconstitution of these deficient mice with the IL-10 gene fully restores development of airway hyperresponsiveness comparable to control mice. These results identify an important role of IL-10, downstream of the inflammatory cascade, in regulating the tone of the airways after allergic sensitization and challenge. PMID- 10811897 TI - NF-kappa B inhibition causes spontaneous apoptosis in Epstein-Barr virus transformed lymphoblastoid cells. AB - Epstein-Barr virus (EBV) transforms B lymphocytes into lymphoblastoid cell lines usurping the Notch and tumor necrosis factor receptor pathways to effect transcription including NF-kappaB activation. To determine whether NF-kappaB activity is essential in the growth and survival of EBV-transformed lymphoblastoid cell lines, a nondegradable IkappaBalpha mutant was expressed under tetracycline regulation. Despite continued Bcl-2 and Bcl-x/L expression, NF kappaB inhibition induced apoptosis as evidenced by poly(ADP-ribose) polymerase cleavage, nuclear condensation and fragmentation, and hypodiploid DNA content. Both caspase 3 and 8 activation and loss of mitochondrial membrane potential were observed in apoptotic cells. However, caspase inhibition failed to block apoptosis. These experiments indicate that NF-kappaB inhibitors may be useful in the therapy of EBV-induced cellular proliferation. PMID- 10811898 TI - Mitochondrial and extramitochondrial apoptotic signaling pathways in cerebrocortical neurons. AB - In cultured cerebrocortical neurons, mild excitotoxic insults or staurosporine result in apoptosis. We show here that N-methyl-d-aspartate (NMDA) receptor mediated, but not staurosporine-mediated, apoptosis is preceded by depolarization of the mitochondrial membrane potential (Deltapsi(m)) and ATP loss. Both insults, however, release cytochrome c (Cyt c) into the cytoplasm. What prompts mitochondria to release Cyt c and the mechanism of release are as yet unknown. We examined the effect of inhibition of the adenine nucleotide translocator (ANT), a putative component of the mitochondrial permeability transition pore. Inhibition of the mitochondrial ANT with bongkrekic acid (BA) prevented NMDA receptor mediated apoptosis of cerebrocortical neurons. Concomitantly, BA prevented Deltapsi(m) depolarization, promoted recovery of cellular ATP content, and blocked caspase-3 activation. However, in the presence of BA, Cyt c was still released. Because BA prevented NMDA-induced caspase-3 activation and apoptosis, the presence of Cyt c in the neuronal cytoplasm is not sufficient for the induction of caspase activity or apoptosis. In contrast to these findings, BA was ineffective in preventing staurosporine-induced activation of caspases or apoptosis. Additionally, staurosporine-induced, but not NMDA-induced, apoptosis was associated with activation of caspase-8. These results indicate that, in cerebrocortical cultures, excessive NMDA receptor activation precipitates neuronal apoptosis by means of mitochondrial dysfunction, whereas staurosporine utilizes a distinct pathway. PMID- 10811899 TI - Nonsaturation of AMPA and NMDA receptors at hippocampal synapses. AB - An important issue in synaptic physiology is the extent to which postsynaptic receptors are saturated by the neurotransmitter released from a single synaptic vesicle. Although the bulk of evidence supports receptor saturation, recent studies have started to reveal that alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) and N-methyl-D-aspartate (NMDA) receptors may not be saturated by a single vesicle of glutamate. Here, we address this question through a study of putative single synapses, made by hippocampal neurons in culture, that are identified by FM1-43 staining. An analysis of the sources of variability in the amplitudes of miniature excitatory postsynaptic currents at single synapses reveals that this variability must arise presynaptically, from variations in the quantity of agonist released. Thus, glutamate receptors at hippocampal synapses are not generally saturated by quantal release. PMID- 10811901 TI - Umbrellas and flagships: efficient conservation surrogates or expensive mistakes? AB - The use of umbrella and flagship species as surrogates for regional biota whose spatial distributions are poorly known is a popular conservation strategy. Yet many assumptions underlying the choice of surrogate species remain untested. By using biodiversity databases containing spatial incidence data for species of concern for (i) the southern California coastal sage scrub habitat, (ii) the Columbia Plateau ecoregion, and (iii) the continental United States, we evaluate the potential effectiveness of a range of conservation surrogate schemes (e.g., big carnivores, charismatic species, keystone species, wide-ranging species), asking how many species potentially are protected by each scheme and at what cost in each habitat area. For all three databases, we find that none of the surrogate schemes we evaluated performs significantly better than do a comparable number of species randomly selected from the database. Although some surrogate species may have considerable publicity value, based on the databases we analyzed, representing diverse taxa on three different geographic scales, we find that the utility of umbrella and flagship species as surrogates for regional biodiversity may be limited. PMID- 10811900 TI - Expression profiling of pancreatic beta cells: glucose regulation of secretory and metabolic pathway genes. AB - Pancreatic beta cells respond to changes in blood glucose by secreting insulin and increasing insulin synthesis. To identify genes used in these responses, we have carried out expression profiling of beta cells exposed to high (25 mM) or low (5.5 mM) glucose by using oligonucleotide microarrays. Functional clustering of genes that averaged a 2.2-fold or greater change revealed large groups of secretory pathway components, enzymes of intermediary metabolism, cell-signaling components, and transcription factors. Many secretory pathway genes were up regulated in high glucose, including seven members of the endoplasmic reticulum (ER) translocon. In agreement with array analysis, protein levels of translocon components were increased by high glucose. Most dramatically, the alpha subunit of the signal recognition particle receptor was increased over 20-fold. These data indicate that the translocon and ribosome docking are major regulatory targets of glucose in the beta cell. Analysis of genes encoding enzymes of intermediary metabolism indicated that low glucose brought about greater utilization of amino acids as an energy source. This conclusion was supported by observations of increased urea production under low-glucose conditions. The above results demonstrate genome-wide integration of beta-cell functions at the level of transcript abundance and validate the efficacy of expression profiling in identifying genes involved in the beta-cell glucose response. PMID- 10811903 TI - Calcium triggers an intramolecular association of the C2 domains in synaptotagmin. AB - Synaptotagmin I is a critical component of the synaptic machinery that senses calcium influx and triggers synaptic vesicle fusion and neurotransmitter release. Fluorescence resonance energy transfer studies conducted on synaptotagmin demonstrate that calcium concentrations required for fusion induce a conformational change (EC(50) approximately 3 mM) that brings the two calcium binding C2 domains in synaptotagmin closer together. Analytical ultracentrifugation studies reveal that synaptotagmin is monomeric under these conditions, indicating that this calcium-triggered association between the C2 domains is intramolecular, rather than intermolecular. These results suggest a mechanism for synaptotagmin function at the presynaptic plasma membrane that involves the self-association of C2 domains. PMID- 10811902 TI - The ADP ribosylation factor nucleotide exchange factor ARNO promotes beta arrestin release necessary for luteinizing hormone/choriogonadotropin receptor desensitization. AB - Desensitization of guanine nucleotide binding protein-coupled receptors is a ubiquitous phenomenon characterized by declining effector activity upon persistent agonist stimulation. The luteinizing hormone/choriogonadotropin receptor (LH/CGR) in ovarian follicles exhibits desensitization of effector adenylyl cyclase activity in response to the mid-cycle surge of LH. We have previously shown that uncoupling of the agonist-activated LH/CGR from the stimulatory G protein (G(s)) is dependent on GTP and attributable to binding of beta-arrestin present in adenylyl cyclase-rich follicular membrane fraction to the third intracellular (3i) loop of the receptor. Here, we report that LH/CGR dependent desensitization is mimicked by ADP ribosylation factor nucleotide binding site opener, a guanine nucleotide exchange factor of the small G proteins ADP ribosylation factors (Arfs) 1 and 6, and blocked by synthetic N-terminal Arf6 peptide, suggesting that the GTP-dependent step of LH/CGR desensitization is receptor-dependent Arf6 activation. Arf activation by GTP and ADP ribosylation factor nucelotide-binding site opener promotes the release of docked beta arrestin from the membrane, making beta-arrestin available for LH/CGR; Arf6 but not Arf1 peptides block beta-arrestin release from the membrane. Thus, LH/CGR appears to activate two membrane delimited signaling cascades via two types of G proteins: heterotrimeric G(s) and small G protein Arf6. Arf6 activation releases docked beta-arrestin necessary for receptor desensitization, providing a feedback mechanism for receptor self-regulation. PMID- 10811904 TI - Femtosecond linear dichroism of DNA-intercalating chromophores: solvation and charge separation dynamics of [Ru(phen)2dppz]2+ systems. AB - The DNA-intercalating chromophore [Ru(phen)(2)dppz](2+) has unique photophysical properties, the most striking of which is the "light-switch" characteristic when binding to DNA. As a dimer, it acts as a molecular staple for DNA, exhibiting a remarkable double-intercalating topology. Herein, we report femtosecond dynamics of the monomeric and the covalently linked dimeric chromophores, both free in aqueous solution and complexed with DNA. Transient absorption and linear dichroism show the electronic relaxation to the lowest metal-to-ligand charge transfer (CT) state, and subpicosecond kinetics have been observed for this chromophore for what is, to our knowledge, the first time. We observe two distinct relaxation processes in aqueous solution with time constants of 700 fs and 4 ps. Interestingly, these two time constants are very similar to those observed for the reorientational modes of bulk water. The 700-fs process involves a major dichroism change. We relate these observations to the change in charge distribution and to the time scales involved in solvation of the CT state. Slower processes, with lifetimes of approximately 7 and 37 ps, were observed for both monomer and dimer when bound to DNA. Such a difference can be ascribed to the change of the structural and electronic relaxation experienced in the DNA intercalation pocket. Finally, the recombination lifetime of the final metal-to ligand CT state to the ground state, which is a key in the light-switch process, is found in aqueous solution to be sensitive to structural modification, ranging from 260 ps for [Ru(phen)(2)dppz](2+) and 360 ps for the monomer chromophore derivative to 2.0 ns for the dimer. This large change reflects the direct role of solvation in the light-switch process. PMID- 10811905 TI - An efficient recombination system for chromosome engineering in Escherichia coli. AB - A recombination system has been developed for efficient chromosome engineering in Escherichia coli by using electroporated linear DNA. A defective lambda prophage supplies functions that protect and recombine an electroporated linear DNA substrate in the bacterial cell. The use of recombination eliminates the requirement for standard cloning as all novel joints are engineered by chemical synthesis in vitro and the linear DNA is efficiently recombined into place in vivo. The technology and manipulations required are simple and straightforward. A temperature-dependent repressor tightly controls prophage expression, and, thus, recombination functions can be transiently supplied by shifting cultures to 42 degrees C for 15 min. The efficient prophage recombination system does not require host RecA function and depends primarily on Exo, Beta, and Gam functions expressed from the defective lambda prophage. The defective prophage can be moved to other strains and can be easily removed from any strain. Gene disruptions and modifications of both the bacterial chromosome and bacterial plasmids are possible. This system will be especially useful for the engineering of large bacterial plasmids such as those from bacterial artificial chromosome libraries. PMID- 10811906 TI - Drosophila Thor participates in host immune defense and connects a translational regulator with innate immunity. AB - Thor has been identified as a new type of gene involved in Drosophila host immune defense. Thor is a member of the 4E-binding protein (4E-BP) family, which in mammals has been defined as critical regulators in a pathway that controls initiation of translation through binding eukaryotic initiation factor 4E (eIF4E). Without an infection, Thor is expressed during all developmental stages and transcripts localize to a wide variety of tissues, including the reproductive system. In response to bacterial infection and, to a lesser extent, by wounding, Thor is up-regulated. The Thor promoter has the canonical NFkappaB and associated GATA recognition sequences that have been shown to be essential for immune induction, as well as other sequences commonly found for Drosophila immune response genes, including interferon-related regulatory sequences. In survival tests, Thor mutants show symptoms of being immune compromised, indicating that Thor may be critical in host defense. In contrast to Thor, Drosophila eIF4E is not induced by bacterial infection. These findings for Thor provide the first evidence that a 4E-BP family member has a role in immune induction in any organism. Further, no gene in the translation initiation pathway that includes 4E BP has been previously found to be immune induced. Our results suggest either a role for translational regulation in humoral immunity or a new, nontranslational function for 4E-BP type genes. PMID- 10811907 TI - Protein oxidation in response to increased transcriptional or translational errors. AB - In this study, we show a correlation between synthesis of aberrant proteins and their oxidative modification. The level of aberrant proteins was elevated in Escherichia coli cultures by decreasing transcriptional or translational fidelity using specific mutations or drugs. Protein carbonylation, an oxidative modification, increased in parallel to the induction of the heat shock chaperone GroEL. As the protein turnover rates and level of intracellular oxidative stress remained unchanged, it appears that carbonylation results from the increased susceptibility of the misfolded proteins. These studies show that the cellular protein oxidation is not limited only by available reactive oxygen species, but by the levels of aberrant proteins. Thus, protein oxidation seen in aging cells may be the consequence also of reduced transcriptional/translational fidelity, and protein structures appear to have evolved to minimize oxidative damage. In addition, we discuss the possibility that carbonylation, being an unrepairable protein modification, may serve as a tagging system to shunt misfolded proteins between pathways of refolding by chaperones or the proteolytic apparatus. PMID- 10811908 TI - Ser67-phosphorylated inhibitor 1 is a potent protein phosphatase 1 inhibitor. AB - Inhibitor 1 (I-1) is a protein inhibitor of protein phosphatase 1 (PP1), a major eukaryotic Ser/Thr phosphatase. Nonphosphorylated I-1 is inactive, whereas phosphorylated I-1 is a potent PP1 inhibitor. I-1 is phosphorylated in vivo on Thr(35) and Ser(67). Thr(35) is phosphorylated by cAMP-dependent protein kinase (A kinase), and Thr(35)-phosphorylated I-1 inhibits PP1. Until now the kinase that phosphorylates Ser(67) had not been identified and the physiological role of Ser(67) phosphorylation was unknown. In this study we detected a high level of kinase activity in brain extract when a glutathione S-transferase (GST) fusion I 1 mutant containing an Ala substituted for Thr(35) [GST-I-1(T35A)] was used as the substrate. GST-I-1(T35A) kinase and neuronal cdc2-like protein kinase (NCLK) in the brain extract could not be separated from each other by a series of sequential chromatographies. GST-I-1(T35A) kinase immunoprecipitated with anti NCLK antibody from kinase-active column fractions. Purified NCLK-phosphorylated GST-I-1(T35A) and I-1 (0.7 mole of phosphate per mole of I-1). HPLC phosphopeptide mapping, amino acid sequencing, and site-directed mutagenesis determined that NCLK phosphorylates Ser(67) of I-1. NCLK-phosphorylated I-1 and I 1(T35A) inhibited PP1 with IC(50) values approximately 9.5 and 13. 8 nM, respectively. When compared, A kinase-phosphorylated I-1 was only approximately 1.2 times more inhibitory than NCLK-phosphorylated I-1. Our data indicate that NCLK is a potential in vivo I-1 kinase and that Thr(35) and Ser(67) phosphorylation independently activate I-1. PMID- 10811909 TI - An essential intermediate in the folding of dihydrofolate reductase. AB - The folding of Escherichia coli dihydrofolate reductase was examined at pH 7.8 and 15 degrees C by using stopped-flow fluorescence and absorbance spectroscopies. The formation of a highly fluorescent intermediate occurs with relaxation times ranging between 142 and 343 msec, whereas stopped-flow absorbance spectroscopy using methotrexate binding assays shows a distinct lag phase during these time frames for the native state. The lag in absorbance kinetics and the lack of fast-track folding events indicate that the formation of this ensemble of intermediates is an obligatory step in the folding reaction. PMID- 10811910 TI - How native-state topology affects the folding of dihydrofolate reductase and interleukin-1beta. AB - The overall structure of the transition-state and intermediate ensembles observed experimentally for dihydrofolate reductase and interleukin-1beta can be obtained by using simplified models that have almost no energetic frustration. The predictive power of these models suggests that, even for these very large proteins with completely different folding mechanisms and functions, real protein sequences are sufficiently well designed, and much of the structural heterogeneity observed in the intermediates and the transition-state ensembles is determined by topological effects. PMID- 10811911 TI - High frequency of hypermethylation at the 14-3-3 sigma locus leads to gene silencing in breast cancer. AB - Expression of 14-3-3 final sigma (final sigma) is induced in response to DNA damage, and causes cells to arrest in G(2). By SAGE (serial analysis of gene expression) analysis, we identified final sigma as a gene whose expression is 7 fold lower in breast carcinoma cells than in normal breast epithelium. We verified this finding by Northern blot analysis. Remarkably, final sigma mRNA was undetectable in 45 of 48 primary breast carcinomas. Genetic alterations at final sigma such as loss of heterozygosity were rare (1/20 informative cases), and no mutations were detected (0/34). On the other hand, hypermethylation of CpG islands in the final sigma gene was detected in 91% (75/82) of breast tumors and was associated with lack of gene expression. Hypermethylation of final sigma is functionally important, because treatment of final sigma-non-expressing breast cancer cell lines with the drug 5-aza-2'-deoxycytidine resulted in demethylation of the gene and synthesis of final sigma mRNA. Breast cancer cells lacking final sigma expression showed increased number of chromosomal breaks and gaps when exposed to gamma-irradiation. Therefore, it is possible that loss of final sigma expression contributes to malignant transformation by impairing the G(2) cell cycle checkpoint function, thus allowing an accumulation of genetic defects. Hypermethylation and loss of final sigma expression are the most consistent molecular alterations in breast cancer identified so far. PMID- 10811912 TI - Squamous epithelial proliferation induced by walleye dermal sarcoma retrovirus cyclin in transgenic mice. AB - Walleye dermal sarcoma (WDS) is a common disease of walleye fish in the United States and Canada. These proliferative lesions are present autumn through winter and regress in the spring. Walleye dermal sarcoma virus (WDSV), a retrovirus distantly related to other members of the family Retroviridae, has been etiologically linked to the development of WDS. We have reported that the D cyclin homologue [retroviral (rv) cyclin] encoded by WDSV rescues yeast conditionally deficient for cyclin synthesis from growth arrest and that WDSV cyclin mRNA is present in developing tumors. These data strongly suggest that the rv-cyclin plays a central role in the development of WDS. To test the ability of the WDSV rv-cyclin to induce cell proliferation, we have generated transgenic mice expressing the rv-cyclin in squamous epithelia from the bovine keratin-5 promoter. The transgenic animals were smaller than littermates, had reduced numbers of hair follicles, and transgenic females did not lactate properly. Following injury the transgenic animals developed severe squamous epithelial hyperplasia and dysplasia with ultrastructural characteristics of neoplastic squamous epithelium. Immunocytochemistry studies demonstrated that the hyperplastic epithelium stained positive for cytokeratin and were abnormally differentiated. Furthermore, the rv-cyclin protein was detected in the thickened basal cell layers of the proliferating lesions. These data are the first to indicate that the highly divergent WDSV rv-cyclin is a very potent stimulator of eukaryotic cell proliferation and to demonstrate the potential of a cyclin homologue encoded by a retrovirus to induce hyperplastic skin lesions. PMID- 10811913 TI - Differential activation of a Candida albicans virulence gene family during infection. AB - The yeast Candida albicans is a harmless commensal in most healthy people, but it causes superficial as well as life-threatening systemic infections in immunocompromised patients. C. albicans can colonize or infect virtually all body sites because of its high adaptability to different host niches, which involves the activation of appropriate sets of genes in response to complex environmental signals. We have used an in vivo expression technology that is based on genetic recombination as a reporter of gene expression to monitor the differential activation of individual members of a gene family encoding secreted aspartic proteinases (Saps), which have been implicated in C. albicans virulence, at various stages of the infection process. Our results demonstrate that SAP expression depends on the type of infection, with different SAP isogenes being activated during systemic disease as compared with mucosal infection. In addition, the activation of individual SAP genes depends on the progress of the infection, some members of the gene family being induced immediately after contact with the host, whereas others are expressed only after dissemination into deep organs. In the latter case, the number of invading organisms determines whether induction of a virulence gene is necessary for successful infection. The in vivo expression technology allows the elucidation of gene expression patterns at different stages of the fungus-host interaction, thereby revealing regulatory adaptation mechanisms that make C. albicans the most successful fungal pathogen of humans and, at the same time, identifying the stage of an infection at which certain virulence genes may play a role. PMID- 10811915 TI - Bruchins: insect-derived plant regulators that stimulate neoplasm formation. AB - Pea weevil (Bruchus pisorum L.) oviposition on pods of specific genetic lines of pea (Pisum sativum L.) stimulates cell division at the sites of egg attachment. As a result, tumor-like growths of undifferentiated cells (neoplasms) develop beneath the egg. These neoplasms impede larval entry into the pod. This unique form of induced resistance is conditioned by the Np allele and mediated by a recently discovered class of natural products that we have identified from both cowpea weevil (Callosobruchus maculatus F.) and pea weevil. These compounds, which we refer to as "bruchins," are long-chain alpha,omega-diols, esterified at one or both oxygens with 3-hydroxypropanoic acid. Bruchins are potent plant regulators, with application of as little as 1 fmol (0.5 pg) causing neoplastic growth on pods of all of the pea lines tested. The bruchins are, to our knowledge, the first natural products discovered with the ability to induce neoplasm formation when applied to intact plants. PMID- 10811914 TI - Highly reduced protection against Streptococcus pneumoniae after deletion of a single heavy chain gene in mouse. AB - Phosphocholine (PC) is the immunodominant epitope found on the surface of Streptococcus pneumoniae (SPn). T15-idiotype Abs, whose heavy (H) chain variable region is encoded by the V1 gene, are dominant in the anti-PC response in adult mice and protect mice from lethal pneumococcal infection. The ability of anti-PC Abs using H chains other than the V1 H chain to protect against pneumococcal infection remains controversial. We generated V1(-/-) knockout mice to determine whether protective anti-PC Abs could be produced in the absence of the V1 gene. No anti-PC Abs were produced in V1(-/-) mice immunized with avirulent SPn; however, PC-BSA binding Abs were induced after immunization with PC-keyhole limpet hemocyanin but at significantly lower levels than those in wild-type mice. These Abs provided poor protection against virulent SPn; thus, <25% of V1(-/-) mice survived challenge with 10(4) bacteria as compared with 100% survival of V1(+/+) mice. The anti-PC Abs in V1(-/-) mice were heteroclitic, binding to nitrophenyl-PC better than to PC. None of nine hybridomas produced from V1(-/-) mice provided passive protection. However, the V1(-/-) mice produced normal amounts of Ab to SPn proteins that can partially protect mice against SPn. These data indicate that the V1 gene is critical for the production of anti-PC Abs providing optimum protection against infection with SPn, and the V1(-/-) mice could be useful in unmasking epitopes other than the immunodominant PC epitope on SPn capable of providing cross protection. PMID- 10811916 TI - brakeless is required for photoreceptor growth-cone targeting in Drosophila. AB - The R1-R6 subclass of photoreceptor neurons (R cells) in the Drosophila compound eye form specific connections with targets in the optic ganglia. In this paper, we report the identification of a gene, brakeless (bks), that is essential for R1 R6 growth cone targeting. In brakeless mutants, R1-R6 growth cones frequently fail to terminate migration in their normal target, the lamina, and instead project through it and terminate in the second optic ganglion, the medulla. Genetic mosaic analysis and transgene rescue experiments indicate that bks functions in R cells and not within the lamina target region. bks encodes a nuclear protein. We propose that it participates in a gene expression pathway regulating one or more growth cone components controlling R1-R6 targeting. PMID- 10811917 TI - Oxytocin and its receptors are synthesized in the rat vasculature. AB - Produced and released by the heart, oxytocin (OT) acts on its cardiac receptors to decrease the cardiac rate and force of contraction. We hypothesized that it might also be produced in the vasculature and regulate vascular tone. Consequently, we prepared acid extracts of the pulmonary artery and vena cava of female rats. OT concentrations in dog and sheep aortae were equivalent to those of rat aorta (2745 +/- 180 pg/mg protein), indicating that it is present in the vasculature of several mammalian species. Reverse-phase HPLC of aorta and vena cava extracts revealed a single peak corresponding to the amidated OT nonapeptide. Reverse-transcribed PCR confirmed OT synthesis in these tissues. Using the selective OT receptor ligand compound VI, we detected a high number of OT-binding sites in the rat vena cava and aorta. Furthermore, OT receptor (OTR) mRNA was found in the vena cava, pulmonary vein, and pulmonary artery with lower levels in the aorta, suggesting vessel-specific OTR distribution. The abundance of OTR mRNA in the vena cava and pulmonary vein was associated with high atrial natriuretic peptide mRNA. In addition, we have demonstrated that diethylstilbestrol treatment of immature female rats increased OT significantly in the vena cava but not in the aorta and augmented OTR mRNA in both the aorta (4 fold) and vena cava (2-fold), implying regulation by estrogen. Altogether, these data suggest that the vasculature contains an intrinsic OT system, which may be involved in the regulation of vascular tone as well as vascular regrowth and remodeling. PMID- 10811918 TI - Secreted and membrane attractin result from alternative splicing of the human ATRN gene. AB - Attractin, initially identified as a soluble human plasma protein with dipeptidyl peptidase IV activity that is expressed and released by activated T lymphocytes, also has been identified as the product of the murine mahogany gene with connections to control of pigmentation and energy metabolism. The mahogany product, however, is a transmembrane protein, raising the possibility of a human membrane attractin in addition to the secreted form. The genomic structure of human attractin reveals that soluble attractin arises from transcription of 25 sequential exons on human chromosome 20p13, where the 3' terminal exon contains sequence from a long interspersed nuclear element-1 (LINE-1) retrotransposon element that includes a stop codon and a polyadenylation signal. The mRNA isoform for membrane attractin splices over the LINE-1 exon and includes five exons encoding transmembrane and cytoplasmic domains with organization and coding potential almost identical to that of the mouse gene. The relative abundance of soluble and transmembrane isoforms measured by reverse transcription-PCR is differentially regulated in lymphoid tissues. Because activation of peripheral blood leukocytes with phytohemagglutinin induces strong expression of cell surface attractin followed by release of soluble attractin, these results suggest that a genomic event unique to mammals, LINE-1 insertion, has provided an evolutionary mechanism for regulating cell interactions during an inflammatory reaction. PMID- 10811919 TI - The ryanodine receptor is essential for larval development in Drosophila melanogaster. AB - We have investigated the role of the ryanodine receptor in Drosophila development by using pharmacological and genetic approaches. We identified a P element insertion in the Drosophila ryanodine receptor gene, Ryanodine receptor 44F (Ryr), and used it to generate the hypomorphic allele Ryr(16). An examination of hypodermal, visceral, and circulatory muscle showed that, in each case, muscle contraction was impaired in Ryr(16) larvae. Treatment with the drug ryanodine, a highly specific modulator of ryanodine receptor channel activity, also inhibited muscle function, and, at high levels, completely blocked hypodermal muscle contraction. These results suggest that the ryanodine receptor is required for proper muscle function and may be essential for excitation-contraction coupling in larval body wall muscles. Nonmuscle roles of Ryr were also investigated. Ryanodine-sensitive Ca(2+) stores had previously been implicated in phototransduction; to address this, we generated Ryr(16) mutant clones in the adult eye and performed whole-cell, patch-clamp recordings on dissociated ommatidia. Our results do not support a role for Ryr in normal light responses. PMID- 10811920 TI - The silencing protein SIR2 and its homologs are NAD-dependent protein deacetylases. AB - Homologs of the chromatin-bound yeast silent information regulator 2 (SIR2) protein are found in organisms from all biological kingdoms. SIR2 itself was originally discovered to influence mating-type control in haploid cells by locus specific transcriptional silencing. Since then, SIR2 and its homologs have been suggested to play additional roles in suppression of recombination, chromosomal stability, metabolic regulation, meiosis, and aging. Considering the far-ranging nature of these functions, a major experimental goal has been to understand the molecular mechanism(s) by which this family of proteins acts. We report here that members of the SIR2 family catalyze an NAD-nicotinamide exchange reaction that requires the presence of acetylated lysines such as those found in the N termini of histones. Significantly, these enzymes also catalyze histone deacetylation in a reaction that absolutely requires NAD, thereby distinguishing them from previously characterized deacetylases. The enzymes are active on histone substrates that have been acetylated by both chromatin assembly-linked and transcription-related acetyltransferases. Contrary to a recent report, we find no evidence that these proteins ADP-ribosylate histones. Discovery of an intrinsic deacetylation activity for the conserved SIR2 family provides a mechanism for modifying histones and other proteins to regulate transcription and diverse biological processes. PMID- 10811921 TI - Interactions between heterologous forms of prion protein: binding, inhibition of conversion, and species barriers. AB - The self-induced formation of the disease-associated, protease-resistant prion protein (PrP-res) from the normal protease-sensitive isoform (PrP-sen) appears to be a key event in the pathogenesis of transmissible spongiform encephalopathies. The amino acid sequence specificity of PrP-res formation correlates with, and may account for, the species specificity in transmission of transmissible spongiform encephalopathy agents in vivo. To analyze the mechanism controlling the sequence specificity of PrP-res formation, we compared the binding of PrP-sen to PrP-res with its subsequent acquisition of protease resistance by using cell-free systems consisting of heterologous versus homologous mouse and hamster PrP isoforms. Our studies showed that heterologous PrP-sen can bind to PrP-res with little conversion to the protease-resistant state and, in doing so, can interfere with the conversion of homologous PrP-sen. The interference occurred with molar ratios of homologous to heterologous PrP-sen molecules as low as 1:1. The interference was due primarily to the inhibition of conversion, but not the binding, of the homologous PrP-sen to PrP-res. The results provide evidence that the sequence specificity of PrP-res formation in this model is determined more by the conversion to protease resistance than by the initial binding step. These findings also imply that after the initial binding, further intermolecular interactions between PrP-sen and PrP-res are required to complete the process of conversion to the protease-resistant state. PMID- 10811922 TI - Neuronal correlates of sensory discrimination in the somatosensory cortex. AB - Monkeys are able to discriminate the difference in frequency between two periodic mechanical vibrations applied sequentially to the fingertips. It has been proposed that this ability is mediated by the periodicity of the responses in the quickly adapting (QA) neurons of the primary somatosensory cortex (S1), instead of the average firing rates. We recorded from QA neurons of S1 while monkeys performed the vibrotactile discrimination task. We found that the periodic mechanical vibrations can be represented both in the periodicity and in the firing rate responses to varying degrees across the QA neuronal population. We then computed neurometric functions by using both the periodicity and the firing rate and sought to determine which of these two measures is associated with the psychophysical performance. We found that neurometric thresholds based on the firing rate are very similar to the animal's psychometric thresholds whereas neurometric thresholds based on periodicity are far lower than those thresholds. These results indicate that an observer could solve this task with a precision similar to that of the monkey, based only on the firing rate produced during the stimulus periods. PMID- 10811923 TI - The many faces of DNA polymerases: strategies for mutagenesis and for mutational avoidance. PMID- 10811924 TI - Functional brain imaging to identify affected subjects genetically at risk for Alzheimer's disease. PMID- 10811925 TI - Cellular receptors for viruses: links to tropism and pathogenesis. PMID- 10811926 TI - CD46 transgene expression in pig peripheral blood mononuclear cells does not alter their susceptibility to measles virus or their capacity to downregulate endogenous and transgenic CD46. AB - CD46 (or membrane cofactor protein) protects autologous cells from complement mediated lysis and has been expressed as a transgene in pigs to overcome complement-mediated hyperacute rejection of porcine organs upon transplantation into primates. Since CD46 has been identified as a receptor for measles virus (MV), the susceptibility of CD46-transgenic (tg) pig peripheral blood mononuclear cells (PBMC) to infection with MV strains which do and do not use CD46 as receptor was investigated. Surprisingly, it was found that MV vaccine strains (e.g. Edmonston) bound to tg as well as non-tg pig PBMC. Phytohaemagglutinin stimulated CD46-tg and non-tg pig PBMC were equally well infected with MV vaccine strains irrespective of CD46 expression. Upon infection, tg CD46 was downregulated from the cell surface. In contrast, the binding capacity for MV wild-type strains to pig and human PBMC was low, irrespective of CD46 expression. These MV strains did not infect tg or non-tg pig cells. Expression of endogenous pig CD46 was detected with polyclonal sera against human CD46. After infection of pig PBMC with MV strain Edmonston, endogenous pig CD46 was also downregulated. This suggests an interaction between MV Edmonston and pig CD46. However, polyclonal CD46 sera did not inhibit infection with MV Edmonston indicating that CD46 may not exclusively act as a receptor for MV on these cells. Interestingly, similar results were observed using human PBMC. Data suggest that CD46 downregulation after interaction with MV may also occur in porcine organs which express endogenous and/or human CD46 as a means of protection against complement mediated damage. PMID- 10811927 TI - Long-term protective immunity to rinderpest in cattle following a single vaccination with a recombinant vaccinia virus expressing the virus haemagglutinin protein. AB - A recombinant vaccine, produced by using a highly attenuated smallpox vaccine (LC16mO) as a vector and which expresses the rinderpest virus (RPV) haemagglutinin protein, has been developed. The properties of this vaccine, including its heat stability, efficacy in short-term trials, safety and genetic stability, have been confirmed in an earlier report. In the present study, the duration of the protective immunity generated by the vaccine in cattle was examined for up to 3 years following the administration of a single vaccination dose of 10(8) p.f.u. The vaccinated cattle were kept for 2 (group I) or 3 years (group II) and then challenged with a highly virulent strain of RPV. Four of five vaccinated cattle in group I and all six cattle in group II survived the challenge, some showing solid immunity without any clinical signs of rinderpest. Neutralizing antibodies were maintained at a significant level for up to 3 years and they increased rapidly following challenge. Lymphocyte proliferative responses to RPV were examined in group II cattle and were observed in four of the six vaccinated cattle in this group. The long-lasting protective immunity, in addition to the other properties confirmed previously, indicate the practical usefulness of this vaccine for field use. PMID- 10811928 TI - Characterization of antigenically unique influenza C virus strains isolated in Yamagata and Sendai cities, Japan, during 1992-1993. AB - Three influenza C virus strains (C/Yamagata/1/92, C/Yamagata/1/93 and C/Miyagi/5/93) isolated in Yamagata and Sendai Cities, Japan, between June 1992 and May 1993 were found to possess haemagglutinin-esterase glycoproteins that were antigenically indistinguishable from one another but were clearly different from any previous Japanese isolates. To investigate the origin of the 1992/1993 strains, their antigenic and genetic properties were compared with those of eight strains isolated outside Japan between 1967 and 1982. The results showed that the 1992/1993 isolates were closely related to a virus isolated in Brazil in 1982 (C/SaoPaulo/378/82) and that these viruses (including C/SaoPaulo/378/82) are reassortants that had obtained PB1 and NP genes from a C/Yamagata/26/81-like parent and the other genes from another as yet unidentified parent. PMID- 10811929 TI - Immunoglobulin A responses to Puumala hantavirus. AB - Puumala hantavirus (PUUV) causes nephropathia epidemica (NE), a form of haemorrhagic fever with renal syndrome that occurs in northern and central Europe. The immunoglobulin A (IgA) response in NE patients was studied. The levels of total serum IgA in acute-phase samples from NE patients were found to be significantly elevated when compared with the levels in healthy controls. ELISAs for detection of the IgA1 and IgA2 responses against each PUUV structural protein (N, G1 and G2) were developed and evaluated. Sequential sera from NE patients (acute, convalescent, 2-year) and 10-20 year NE-convalescent sera were examined. Most patients developed detectable levels of IgA1 against N and G2, while the G1 responses were low or undetectable. Seven of nine 10-20 year sera contained virus-specific IgA1, which may indicate the prolonged presence of viral antigens after the initial infection. PEPSCAN analysis revealed several IgA reactive antigenic regions in the N protein. Serum IgA and IgG was purified by affinity chromatography and examined by a virus-neutralization assay. Three of five sera from acute-phase NE patients contained neutralizing IgA1. The diagnostic potential of the PUUV-specific IgA1 response was evaluated. The N and G2 assays showed specificities of 100% with sensitivities of 91 and 84%, respectively, compared with an IgM mu-capture ELISA. Several NE patients, clinically diagnosed for acute PUUV infection, with borderline or undetectable levels of PUUV-specific IgM, were found to be highly positive for the presence of PUUV N-specific serum IgA1, proving the diagnostic value of IgA analysis as a complement to detection of IgM. PMID- 10811930 TI - Antigenic and genetic stability of bovine immunodeficiency virus during long-term persistence in cattle experimentally infected with the BIV(R29) isolate. AB - Experimental infection of cattle with bovine immunodeficiency virus (BIV) is characterized by persistent, low levels of virus replication in the absence of clinical disease. A virus neutralization (VN) assay was developed to examine the role of VN antibodies in controlling virus replication in cattle experimentally infected with the BIV(R29) isolate of BIV. All animals developed VN antibody, but there was no correlation between VN titres and restriction of virus replication in vivo. BIV infection did not induce high-titred, cross-neutralizing antibody and there was no evidence for antigenic variation through more than 4 years in vivo. Genetic comparisons among the BIV(R29) inoculum virus and viruses isolated from infected animals identified only limited genetic variation during 4 years in vivo. Moreover, there was no evidence that the observed variation was due to selection. Analyses of genetic diversity in the virus stock used for inoculation indicated a fairly homogeneous population. In the absence of high levels of virus replication and overt clinical disease, there appeared to be little selection of virus variants, resulting in antigenic and genetic stability of BIV(R29) during long-term, persistent infection. PMID- 10811931 TI - Gill-associated virus of Penaeus monodon prawns: an invertebrate virus with ORF1a and ORF1b genes related to arteri- and coronaviruses. AB - A 20089 nucleotide (nt) sequence was determined for the 5' end of the (+)-ssRNA genome of gill-associated virus (GAV), a yellow head-like virus infecting Penaeus monodon prawns. Clones were generated from a approximately 22 kb dsRNA purified from lymphoid organ total RNA of GAV-infected prawns. The region contains a single gene comprising two long overlapping open reading frames, ORF1a and ORF1b, of 4060 and 2646 amino acids, respectively. The ORFs are structurally related to the ORF1a and ORF1ab polyproteins of coronaviruses and arteriviruses. The 99 nt overlap between ORF1a and ORF1b contains a putative AAAUUUU 'slippery' sequence associated with -1 ribosomal frameshifting. A 131 nt stem-loop with the potential to form a complex pseudoknot resides 3 nt downstream of this sequence. Although different to the G/UUUAAAC frameshift sites and 'H-type' pseudoknots of nidoviruses, in vitro transcription/translation analysis demonstrated that the GAV element also facilitates read-through of the ORF1a/1b junction. As in coronaviruses, GAV ORF1a encodes a 3C-like cysteine protease domain located between two hydrophobic regions. However, its sequence suggests some structural relationship to the chymotrypsin-like serine proteases of arteriviruses. ORF1b encodes homologues of the 'SDD' polymerase, which among (+)-RNA viruses is unique to nidoviruses, as well as metal-ion-binding and helicase domains. The presence of a dsRNA replicative intermediate and ORF1a and ORF1ab polyproteins translated by a-1 frameshift suggests that GAV represents the first invertebrate member of the Order NIDOVIRALES: PMID- 10811932 TI - A novel hepatitis C virus (HCV) subtype from Somalia and its classification into HCV clade 3. AB - Hepatitis C virus (HCV) sequences from throughout the world have been grouped into six clades, based on recently proposed criteria. Here, the partial sequences and clade assignment are reported for three HCV isolates from chronic hepatitis C patients from Somalia, for whom conventional assays failed to identify the genotype. Phylogenetic analysis of the sequences of the core, envelope 1 and part of the non- structural 5b regions suggests that all three isolates belong to a distinct HCV genetic group, tentatively classified as subtype 3h. This novel HCV subtype shows the highest sequence similarity with HCV isolates from Indonesia. Despite the fact that these patients were infected with HCV clade 3, none of them responded to standard interferon treatment. PMID- 10811933 TI - A multiply substituted G-H loop from foot-and-mouth disease virus in complex with a neutralizing antibody: a role for water molecules. AB - The crystal structure of a 15 amino acid synthetic peptide, corresponding to the sequence of the major antigenic site A (G-H loop of VP1) from a multiple variant of foot-and-mouth disease virus (FMDV), has been determined at 2.3 A resolution. The variant peptide includes four amino acid substitutions in the loop relative to the previously studied peptide representing FMDV C-S8c1 and corresponds to the loop of a natural FMDV isolate of subtype C(1). The peptide was complexed with the Fab fragment of the neutralizing monoclonal antibody 4C4. The peptide adopts a compact fold with a nearly cyclic conformation and a disposition of the receptor-recognition motif Arg-Gly-Asp that is closely related to the previously determined structure for the viral loop, as part of the virion, and for unsubstituted synthetic peptide antigen bound to neutralizing antibodies. New structural findings include the observation that well-defined solvent molecules appear to play a major role in stabilizing the conformation of the peptide and its interactions with the antibody. Structural results are supported by molecular dynamic simulations. The multiply substituted peptide developed compensatory mechanisms to bind the antibody with a conformation very similar to that of its unsubstituted counterpart. One water molecule, which for steric reasons could not occupy the same position in the unsubstituted antigen, establishes hydrogen bonds with three peptide amino acids. The constancy of the structure of an antigenic domain despite multiple amino acid substitutions has implications for vaccine design. PMID- 10811934 TI - Complete sequence determination and genetic analysis of Banna virus and Kadipiro virus: proposal for assignment to a new genus (Seadornavirus) within the family Reoviridae. AB - Arboviruses with genomes composed of 12 segments of double-stranded (ds) RNA have previously been classified as members or probable members of the genus Coltivirus within the family REOVIRIDAE: A number of these viruses have been isolated in North America and Europe and are serologically and genetically related to Colorado tick fever virus, the Coltivirus type species. These isolates constitute subgroup A of the coltiviruses. The complete genome sequences are now presented of two Asian arboviruses, Kadipiro virus (KDV) and Banna virus (BAV), which are currently classified as subgroup B coltiviruses. Analysis of the viral protein sequences shows that all of the BAV genome segments have cognate genes in KDV. The functions of several of these proteins were also indicated by this analysis. Proteins with dsRNA-binding domains or with significant similarities to polymerases, methyltransferases, NTPases or protein kinases were identified. Comparisons of amino acid sequences of the conserved polymerase protein have shown that BAV and KDV are only very distantly related to the subgroup A coltiviruses. These data demonstrate a requirement for the subgroup B viruses to be reassigned to a separate new genus, for which the name Seadornavirus is proposed. PMID- 10811935 TI - Cervical lesions are associated with human papillomavirus type 16 intratypic variants that have high transcriptional activity and increased usage of common mammalian codons. AB - Human papillomavirus type 16 (HPV-16) is a major cause of cervical neoplasia, but only a minority of HPV-16 infections result in cancer. Whether particular HPV-16 variants are associated with cervical disease has not yet been clearly established. An investigation of whether cervical neoplasia is associated with infection with HPV-16 intratypic variants was undertaken by using RFLP analyses in a study of 100 HPV-16 DNA-positive women with or without neoplasia. RFLP variant 2 was positively associated [odds ratio (OR)=2.57] and variant 5 was negatively associated with disease (OR=0.2). Variant 1, which resembles the reference isolate of HPV-16, was found at a similar prevalence among those with and without neoplasia. Variants 1 and 2 were also more likely to be associated with detectable viral mRNA than variant 5 (respectively P=0.03 and P=0.00). When HPV-16 E5 ORFs in 50 clones from 36 clinical samples were sequenced, 19 variant HPV-16 E5 DNA sequences were identified. Twelve of these DNA sequences encoded variant E5 amino acid sequences, 10 of which were novel. Whilst the associations between HPV-16 E5 RFLP variants and neoplasia could not be attributed to differences in amino acid sequences, correlation was observed in codon usage. DNA sequences of RFLP variant 2 (associated with greatest OR for neoplasia) had a significantly greater usage of common mammalian codons compared with RFLP pattern 1 variants. PMID- 10811936 TI - Discrimination of different subsets of cytolytic cells in pseudorabies virus immune and naive pigs. AB - We previously observed that pseudorabies virus (PRV)-induced, cell-mediated cytolysis in pigs includes killing by natural killer (NK) cells. We also observed that IL-2 stimulation in vitro of naive PBMC expands porcine NK cells. The purpose of this study was to compare the phenotypes of the cytolytic subsets stimulated in vitro by PRV and by IL-2. PBMC were isolated from blood of PRV immune and naive pigs and stimulated in vitro with PRV or IL-2. After 6 days, the frequency of various lymphocyte subsets in these cultured PBMC was determined by flow cytometry: the cells were separated with a magnet-activated cell sorter and the cytolytic activity of the separated populations was determined. When lymphocytes were separated and analysed with FACScan, the following lymphocyte subsets were discriminated: CD6(+) CD8(bright+) CD4(-) (CTL phenotype), CD6(+) CD8(dull+) CD4(+) (the fraction containing memory T helper cells), CD6(+) CD8(-) CD4(+) (T helper cell phenotype), CD6(-) CD8(dull+) CD4(-) gammadelta-T(+) ( gammadelta-T cell phenotype), CD6(-) CD8(dull+) CD4(-) gammadelta-T(-) (NK phenotype) and CD6(-) CD8(-) CD4(-) gammadelta-T(-) or gammadelta-T(+). Flow cytometry analysis demonstrated that PRV stimulation of immune PBMC resulted in the occurrence of more CD6(+) CD8(+) and CD4(+) CD8(+) and fewer CD6(-) CD8(+) and gammadelta-T(+) CD8(+) lymphocytes than IL-2 stimulation of naive PBMC (P<0.05). It was demonstrated further that killing by PRV-stimulated PBMC was mediated mainly by CD6(+) CD8(+) T lymphocytes. Killing by IL-2-stimulated PBMC was mediated mainly by CD6(-) CD8(+) T lymphocytes. These results demonstrate that both natural killing and killing by classical PRV-specific CTL were detected in PRV-immune pigs, whereas IL-2 stimulation of PBMC isolated from naive pigs mainly induced natural killing. PMID- 10811937 TI - Characterization of the replication origin (Ori(S)) and adjoining parts of the inverted repeat sequences of the pseudorabies virus genome. AB - The DNA sequence of a 2.4 kbp fragment located in the internal and terminal inverted repeat sequences of the pseudorabies virus genome determined in this study closes a gap between the previously described genes for the ICP4 and ICP22 homologues. The novel sequence contains no conserved herpesvirus open reading frames. Northern blot and cDNA analyses revealed a viral immediate-early transcript of 1.8 kb, which is spliced by the removal of two small introns close to its 5' end and which presumably represents the mRNA of the downstream open reading frame encoding the ICP22 homologue. Upstream of the transcribed region, an imperfect set of three directly repeated sequences was identified. Each of them contains a complementary pair of the alphaherpesvirus origin-binding protein recognition motif GTTCGCAC, spaced by AT-rich sequences. In vitro studies confirmed that the DNA fragment analysed includes a functional origin of viral DNA replication. PMID- 10811938 TI - Characterization of interaction of gH and gL glycoproteins of varicella-zoster virus: their processing and trafficking. AB - Varicella-zoster virus (VZV) glycoproteins gH and gL were examined in a recombinant vaccinia virus system. Single expression of glycoprotein gL produced two molecular forms: an 18 kDa form and a 19 kDa form differing in size by one endoglycosidase H-sensitive N-linked oligosaccharide. Coexpression of gL and gH resulted in binding of the 18 kDa gL form with the mature form of gH, while the 19 kDa gL form remained uncomplexed. The glycosylation processing of gL was not dependent on gH; however, gL was required for the conversion of precursor gH (97 kDa) to mature gH (118 kDa). Subsequent analyses indicated that gL (18 kDa) was a more completely processed gL (19 kDa). Screening of the culture media revealed that gH and gL were secreted, but only if coexpressed and complexed together. The secreted form of gL was 18 kDa while that of gH was 114 kDa. The fact that secreted gH was smaller than intracytoplasmic gH suggested a proteolytic processing event prior to secretion. The 19 kDa form of gL was never secreted. These findings support a VZV gL recycling pathway between the endoplasmic reticulum and the cis-Golgi apparatus. PMID- 10811940 TI - Resistance to TGF-beta1 correlates with a reduction of TGF-beta type II receptor expression in Burkitt's lymphoma and Epstein-Barr virus-transformed B lymphoblastoid cell lines. AB - The pleiotropic cytokine TGF-beta1 is a member of a large family of related factors involved in controlling cell proliferation, differentiation and apoptosis. TGF-beta ligands interact with a complex of type I and type II transmembrane serine/threonine kinases and they transmit their signals to the nucleus via a family of Smad proteins. A panel of over 20 Burkitt's lymphoma (BL) cell lines has been compiled including those that are Epstein-Barr virus (EBV) negative, those that carry EBV with a restricted pattern of EBV latent gene expression (group I) and those that express the full range of latent EBV genes (group III), together with selected EBV-transformed lymphoblastoid cell lines (LCLs). Most of the EBV-negative and group I BL cell lines underwent apoptosis or a G(1) arrest in response to TGF-beta1 treatment. In contrast, group III cell lines and LCLs were completely refractory to these effects of TGF-beta1. All of the cell lines expressed the TGF-beta pathway Smads and the TGF-beta type I receptor. Lack of responsiveness to TGF-beta1 appears to correlate with a down regulation of TGF-beta type II receptor expression. Studies of EBV-converted and stably transfected BL cell lines demonstrated that the EBV gene LMP-1 is neither necessary nor sufficient to block the TGF-beta1 response. PMID- 10811939 TI - Human cytomegalovirus mediates cell cycle progression through G(1) into early S phase in terminally differentiated cells. AB - Terminal differentiation of embryonal carcinoma cells and monocytes has been shown to be important for their permissiveness for human cytomegalovirus (HCMV) infection, even though such terminally differentiated cells have withdrawn from the cell cycle and are, essentially, in G(0) arrest. Recently, data from a number of laboratories have shown that productive infection with HCMV of quiescent fibroblasts held reversibly in G(0) of the cell cycle can result in cell cycle progression, which results eventually in cycle arrest. In contrast to quiescent fibroblasts, the effect of HCMV on cells that have withdrawn irreversibly from the cell cycle due to terminal differentiation has not, so far, been addressed. Here, it is shown that, in cells that have arrested in G(0) as a result of terminal differentiation, HCMV is able to induce cell functions associated with progression of the cell cycle through G(1) into early S phase. This progression is correlated with a direct physical and functional interaction between the HCMV 86 kDa major immediate-early protein (IE86) and the cyclin-dependent kinase inhibitor p21(Cip1). PMID- 10811941 TI - Characterization and genomic analysis of tobacco vein clearing virus, a plant pararetrovirus that is transmitted vertically and related to sequences integrated in the host genome. AB - A previously undescribed caulimo-like virus was identified in the hybrid tobacco species Nicotiana edwardsonii, and was named tobacco vein clearing virus (TVCV) after the symptoms associated with its occurrence in this plant. The virions of TVCV are 50 nm in diameter and are composed of a 45 kDa capsid protein and a 7767 bp dsDNA genome. Each strand of the genome is interrupted by a site-specific discontinuity. In genome sequence and arrangement of ORFs TVCV was most similar to cassava vein mosaic virus, indicating that TVCV is a pararetrovirus. No serological relationship was detected between TVCV and any other caulimoviruses, including petunia vein clearing virus, which has similar biological properties. In N. edwardsonii TVCV was seed-transmitted to 100% of progeny plants, but was not transmitted by mechanical inoculation, grafting or Myzus persicae to any of seven other Nicotiana spp. Genomic DNA of TVCV hybridized to genomic DNA of N. edwardsonii and of N. glutinosa, its male parent, but not to genomic DNA of N. clevelandii, the female parent. TVCV has 78% sequence identity with pararetrovirus-like sequences that are present in high copy number in the N. tabacum genome, and TVCV genomic DNA hybridized to genomic DNA of N. tabacum and N. rustica. These observations suggest that the episomal form of TVCV may arise from integrated pararetroviral elements present in N. edwardsonii, that these integrants were inherited from the male parent N. glutinosa, and that these elements are related but not identical to pararetroviral elements occurring in other Nicotiana spp. PMID- 10811942 TI - The GCD10 subunit of yeast eIF-3 binds the methyltransferase-like domain of the 126 and 183 kDa replicase proteins of tobacco mosaic virus in the yeast two hybrid system. AB - The tobacco mosaic virus (TMV) replicase complex contains virus- and host-encoded proteins. In tomato, one of these host proteins was reported previously to be related serologically to the GCD10 subunit of yeast eIF-3. The yeast two-hybrid system has now been used to show that yeast GCD10 interacts selectively with the methyltransferase domain shared by the 126 and 183 kDa TMV replicase proteins. These findings are consistent with a role for a GCD10-like protein in the TMV replicase complex and suggest that, in TMV-infected cells, the machinery of virus replication and protein synthesis may be closely connected. PMID- 10811943 TI - The Trichoplusia ni granulovirus helicase is unable to support replication of Autographa californica multicapsid nucleopolyhedrovirus in cells and larvae of T. ni. AB - Baculovirus DNA helicases are essential for replication and are determinants of host range. Helicases of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Trichoplusia ni granulovirus (TnGV) differ markedly, although both viruses replicate efficiently in the cabbage looper, T. ni. To determine whether the TnGV helicase (P137) could support replication of AcMNPV in T. ni cells or larvae, the native AcMNPV helicase gene (p143) was disrupted and substituted with p137. P137 did not support replication when synthesized by the P143-deficient AcMNPV. Moreover, P137 did not inhibit AcMNPV replication when co-synthesized in the presence of the AcMNPV P143. These results suggest that although TnGV and AcMNPV replicate efficiently in T. ni, specific protein-protein or protein-DNA interactions between baculoviral helicases and viral-specific factors which form the replicase complex are required for virus replication. A novel and rapid method for disrupting AcMNPV genes in E. coli using the commercial Bac-to-Bac AcMNPV baculovirus expression vector is described. PMID- 10811944 TI - DNA-independent ATPase activity of the Trichoplusia ni granulovirus DNA helicase. AB - DNA helicases of baculoviruses are essential for virus replication and have been implicated as molecular determinants of host range. Although these proteins contain seven motifs (I, Ia, II-VI) characteristic of DNA helicases, the two most important characteristics of helicases - duplex-DNA unwinding and ATPase activity - have not been demonstrated. In the present study, a recombinant putative DNA helicase (rP137) of Trichoplusia ni granulovirus (TnGV) was purified from insect cells infected with a recombinant Autographa californica multicapsid nucleopolyhedrovirus that overproduced rP137. The rP137 protein exhibited an intrinsic DNA-independent ATPase activity that required Mg(2+) as a co-factor, an activity that was reduced in the presence of TnGV and phage lambda DNAs. These results provide further evidence that baculovirus helicase genes encode proteins with biochemical properties similar to those of classical DNA helicases. PMID- 10811945 TI - Protein requirements for assembly of virus-like particles of Junonia coenia densovirus in insect cells. AB - The coding sequences of four overlapping polypeptides starting at four different in-frame AUG codons and co-terminating at the stop codon of the cap gene of Junonia coenia densovirus (JcDNV) were inserted under the control of the p10 promoter of Autographa californica nucleopolyhedrovirus (AcMNPV) to generate AcMNPV-VP1 (four polypeptides), AcMNPV-VP2 (three polypeptides), AcMNPV-VP3 (two polypeptides), and AcMNPV-VP4 (one polypeptide) recombinant viruses. In all cases, infection of Spodoptera frugiperda cells (Sf9) by each of the four recombinant viruses resulted in the production of virus-like particles (VLPs) 22 25 nm in diameter. The VLPs produced by the three recombinants AcMNPV-VP2, AcMNPV VP3 and AcMNPV-VP4 were abundant and contained three, two and one polypeptides, respectively. VP4, the shortest polypeptide, thus appears to be sufficient for assembly of VLPs morphologically similar to those formed with two to four polypeptides. The ratio of VPs did not appear to be critical for assembly of the particles. The polypeptide starting at the first AUG immediately downstream from the p10 promoter was always the most abundantly expressed in infected cells, regardless of the construct. In contrast, plaque-purified AcMNPV-VP1 recombinants were unstable and produced less than one-twentieth of the VLPs produced by the others. All VP transcripts started at the TAAG late motif of the p10 promoter and had a poly(A) tail 14 nt downstream of a poly(A) addition signal located 98 nucleotides downstream of the common stop codon. No significant transcription initiation inside the cap sequence of AcMNPV-VP2, AcMNPV-VP3 and AcMNPV-PV4 was observed. PMID- 10811946 TI - Characterization of the murine BSE infectious agent. AB - Bovine spongiform encephalopathy (BSE) is a prion-associated disease where the infectious agent is thought to be a host-encoded protein with a protease resistant conformation (PrP(Sc)). Here, data are presented on the solubilization of purified murine BSE material, using guanidine-HCl as a denaturing agent. This treatment led to loss of infectivity, which was partially recovered on renaturation after dialysis to remove the chaotropic agent. The renatured product was then fractionated on an isopycnic sucrose-density gradient and the fractions were analysed for the presence of PrP(Sc), nucleic acids and infectivity. It was found that the major part of PrP(Sc) (>90%) and the endogenous nucleic acids did not contribute towards the formation of infectious particles on renaturation. Infectivity was distributed in the top three, low-density fractions. Among these, the presence of considerable infectivity in the fraction of lowest density, with barely detectable PrP(Sc), is of particular interest. PMID- 10811947 TI - Molecular analysis of Irish sheep scrapie cases. AB - Different strains of transmissible spongiform encephalopathies in humans and rodent models are associated with the accumulation of PrP(Sc) of distinct molecular characteristics. These characteristics include glycosylation profiles, fragment sizes and long-term resistance of PrP(Sc) to proteinase K. The first objective of this study was to determine the applicability of these criteria to characterize and differentiate sheep scrapie PrP(Sc) and bovine spongiform encephalopathy (BSE) PrP(Sc). PrP(Sc) in sheep scrapie samples from Ireland had clearly distinct molecular characteristics to PrP(Sc) in cattle BSE samples using a monoclonal antibody (MAb P4) directed to position 89-104 of ovine PrP using either brain homogenates or semi-purified scrapie-associated fibrils. Similar glycoprofiles were found when analysing scrapie PrP(Sc) in six different CNS regions (thoracic spinal cord, thalamus, basal ganglia, mediobasal hypothalamus, medulla oblongata and cortex). While the long-term resistance results using a different monoclonal antibody (raised to ruminant PrP positions 145-163; MAb L42) were similar to the results obtained with MAb P4, different glycotyping results were obtained. Given the variation in glycosylation patterns using different antibodies, we conclude that standardization of methodology and antibodies is crucial to the applicability of molecular analysis of ruminant BSE and scrapie samples. PMID- 10811948 TI - alpha6 integrin is not the obligatory cell receptor for bovine papillomavirus type 4 PMID- 10811950 TI - GFP as a Genetic Marker Scorable Throughout the Life Cycle of Transgenic Zebra Fish. AB - A fish expression vector, FRM, was constructed by fusing the carp beta-actin promoter and first intron to the ocean pout antifreeze protein terminator and putative boundary element. Mutant forms of the green fluorescent protein (GFP) were engineered into this vector, and the resultant series of vectors, FRMwg, FRM3wg (green GFP), and FRM2bl (blue GFP), were used to make transgenic zebra fish. After microinjection of either supercoiled or linearized DNA into one celled eggs, GFP-expressing cells could be monitored by fluorescence microscopy commencing with the midblastula transition and continuing through embryogenesis. From adult fish, which retained scorable GFP either as patches or as a uniform fluorescence, 11 green and 1 blue GFP-expressing lines of zebra fish have been established. Expression of GFP was nearly ubiquitous and similar among all of these lines. Embryonic expression could be scored at 15 to 30 hours postfertilization and was seen throughout the embryo with the exceptions of the yolk, red blood cells, and in some lines, portions of the head. Adult expression was in a majority of tissues (e.g., muscle, brain, intestine, and heart, but not red blood cells). The notable difference between lines was that fluorescent eggs were scorable in seven of the lines. Adult homozygotes from a different subset of eight lines could be selected by the relative intensity of the GFP marking when compared with that in sibling heterozygotes. All 12 lines contain apparent single locus, multicopy, tandem integrations (1.5-100 copies per cell) of the transgenic DNA. The fish expression vector FRM could be used to drive nearly ubiquitous and strong expression of gene products other than GFP. The GFP expression vectors, FRMwg, FRM2wg, FRM3wg, and FRM2bl, may be useful for optimization of transgenesis and as a comarker. GFP-expressing zebra fish lines could facilitate experimental analysis of chimerism and in vivo gene targeting. PMID- 10811949 TI - A possible role for alpha-crystallins in lens epithelial cell differentiation. AB - PURPOSE: To test the hypothesis that crystallin proteins may actively participate in the differentiation of lens epithelial cells into fiber cells. METHODS: Primary epithelial cells from adult bovine lenses were cultured at 37 degrees C until reaching 95-100% confluency in approximately 4-7 days. Using osmotic lysis of pinocytic vesicles, cells were then loaded with proteins labeled or unlabeled with the fluorescent marker Texas Red (TR). Fetal bovine proteins loaded into cells were lens water soluble fractions, HPLC-purified alpha-, beta-, and gamma crystallin fractions, or bovine serum albumin (BSA). Cultures were then monitored for morphological changes over a 7 day period. RESULTS: Both TR-labeled and unlabeled water-soluble and a-crystallin fractions from bovine lenses resulted in morphological changes to epithelial cells during the first two h postloading. These changes included aggregation of epithelial cells into raised multilayered cell masses, as well as several cells losing attachment to the dish. The initial changes were subsequently followed by elongation of cells within the mass and an increase in size of the mass, so that by 4 days postloading the multilayered, multicellular structures could be visualized with the unaided eye. Differentiation was confirmed within these structures by expression of MIP 26, beta- and gamma-crystallin. These changes did not occur in cultures containing cells originally loaded with beta- or gamma-crystallin fractions, or with cells loaded with BSA. CONCLUSIONS: The results support the hypothesis that alpha crystallins may actively participate in the differentiation of lens epithelial cells into fiber cells. PMID- 10811951 TI - The Penaeus monodon Chitinase 1 Gene Is Differentially Expressed in the Hepatopancreas During the Molt Cycle. AB - We have isolated a full-length chitinase complementary DNA from the tiger shrimp Penaeus monodon that encodes a 621 amino acid protein possessing the functional domains of the chitinase protein family. The Penaeus monodon chitinase 1 (PmChi 1) gene product is 81.8% identical to a chitinase 1 protein expressed in the hepatopancreas of Penaeus japonicus. Analysis by reverse transcription-polymerase chain reaction (RT-PCR) indicates that PmChi-1 messenger RNA is detectable in the hepatopancreas and the gut. PmChi-1 expression during the molt cycle fluctuates markedly, with lowest mRNA levels at stages A(1), C, and D(3); there is a dramatic increase in transcript abundance at the D(2) stage. Using the same tissues and molt stages, RT-PCR analyses of genes encoding other digestive enzymes (trypsin, chymotrypsin, and cathepsin L), a muscle structural protein (tropomyosin II), and housekeeping proteins (elongation factor II and GTP-binding protein) indicate that PmChi-1 is expressed in a distinct tissue-specific and stage-specific manner. The other digestive enzyme genes are expressed in a similar spatiotemporal pattern, but none exhibited a dramatic increase in transcript abundance at stage D(2). Increased expression of PmChi-1 at D(2) suggests that hepatopancreas-expressed chitinase is involved in the degradation of endogenous chitin in the gut peritrophic membrane prior to molting. PMID- 10811952 TI - Histological Characterization and Glucose Incorporation into Glycogen of the Pacific Oyster Crassostrea gigas Storage Cells. AB - In order to investigate glycogen metabolism in the oyster Crassostrea gigas, the distribution of storage cells in the whole animal was studied before histological and biochemical characterization. These cells were found mainly in the labial palps, the mantle, and gonadal area and also in gills and the digestive area. Storage cells from palps, mantle, and gonad presented the same morphological features and the same seasonal glycogen variations. Storage cells were isolated from the labial palps and the mantle plus gonadal area of the oyster by enzymatic dispersion and centrifugation through discontinuous Percoll gradient. These cells have a modal density of 1.043 g/ml. An ultrastructural study confirmed that glycogen is present in the cytoplasm either as fine particles or sequestered within vesicles. Glucose incorporation into glycogen was evaluated in vitro using [U-(14)C]glucose: the incorporation in isolated cells increased linearly for at least 8 hours, was proportional to the cell concentration, and showed saturation kinetics with respect to the exogenous glucose concentration. PMID- 10811953 TI - Universal Primers for Amplification of Mitochondrial Small Subunit Ribosomal RNA Encoding Gene in Scleractinian Corals. AB - We describe the construction of polymerase chain reaction primers designed to amplify a portion of the mitochondrial (mt) small subunit ribosomal (SSU) RNA encoding genes in scleractinian corals. Combinations of cloning and sequencing show that the amplified fragments are between 694 and 896 bp in length. Alignment of the amplified DNA sequences to the published mt SSU rRNA genes of Metridium senile and Sarcophyton glaucum indicates several conserved regions among actiniarian, corallimorpharian, octocorallian, and scleractinians, suggesting this primer set can successfully amplify over 80% of the mt SSU rDNA region of scleractinian corals. Surveys of sequence variation and estimation of the rate of evolution show an extremely slow divergence of the SSU rRNA gene in the family Acroporidae. PMID- 10811954 TI - Structural Organization of the Atlantic Croaker Connexin 32.2 Gene and Its 5' Flanking Region. AB - Gonadotropic hormone stimulates the accumulation of connexin (Cx) 32.2 messenger RNA in ovaries of the marine teleost Atlantic croaker. This effect can be mimicked by protein kinase A (PKA) activators and blocked by PKA inhibitors as well as protein kinase C (PKC) stimulators. However, the mechanisms of Cx32.2 gene regulation are unknown. In this study, we determined the structure of the Cx32.2 gene as a first step toward characterizing the regulatory mechanisms of Cx32.2 gene expression. A cosmid library of croaker genomic DNA was screened with a Cx32.2 complementary DNA probe. One positive clone was subcloned and sequenced. The Cx32.2 gene contained two exons and one intron. The first exon contained a portion of the 5' untranslated region (UTR), and the second exon contained the remaining 5' UTR, the amino acid coding region, and the 3' UTR. The distal 5' flanking region also contained a sequence homologous to a different croaker Cx gene, Cx32.7, but it is unclear if this sequence constitutes a pseudogene or an exon of the Cx32.7 gene. The 5' flanking region of the Cx32.2 gene contained two core cyclic AMP response elements (CRE, CGTCA) and one full-consensus activating protein- (AP)-1 binding site (AGTCAG). The distal core CRE was associated with a sequence that enhances CRE activity (GAGC). A third core CRE site was present in the intron. These findings are consistent with the following hypotheses: the induction of ovarian Cx32.2 mRNA levels by gonadotropic hormone is at least partly mediated by CRE-dependent activation of the Cx32.2 gene, and the inhibition of basal and gonadotropic-hormone-stimulated ovarian Cx32.2 mRNA by PKC is due to negative effects on transcription via the AP-1 transcription factor complex. This study is the first to characterize the structure and putative response elements of the 5' flanking region of fish Cx genes. PMID- 10811955 TI - The Internally Self-fertilizing Hermaphroditic Teleost Rivulus marmoratus (Cyprinodontiformes, Rivulidae) beta-Actin Gene: Amplification and Sequence Analysis with Conserved Primers. AB - To determine the ease and feasibility of amplifying the beta-actin gene in fish by the polymerase chain reaction (PCR), genomic DNAs of several fish (Rivulus, Southern top mouth minnow, common fat minnow, oily bitterling, carp, Far Eastern catfish, medaka, and European flounder) were extracted and used as a template with conserved primers, designed on the basis of high amino acid homology (approximately 98% or more). Among them, the self-fertilizing hermaphroditic fish Rivulus marmoratus was chosen for further characterization. After amplification of the Rivulus beta-actin PCR product with Taq polymerase, PCR product was subcloned to pCRII vector. After restriction enzyme mapping of Rivulus beta-actin gene, the amplified insert was sequenced using ALF Express automatic DNA sequencer with conserved internal primers. The R. marmoratus beta-actin gene consists of 1763 bp encoding 375 amino acids including 5 exons and 4 introns. The splicing and acceptance sites of the exon and intron boundaries of the Rivulus beta-actin gene were highly conserved with consensus sequences (GT/AG). The amino acid homology of R. marmoratus beta-actin to other species was high: 98.93% to human; 98.93%, Atlantic salmon; 98.93%, common carp; 98.93%, grass carp; 98.93%, zebrafish; 98.67%, medaka; and 98.40%, sea bream. To determine the expression of the R. marmoratus beta-actin gene in liver and ovary, reverse transcriptase polymerase chain reaction was carried out with internal primers. In conclusion, these universal primers are successful in the rapid cloning of the fish beta actin gene by PCR, based on a high homology of the beta-actin gene conserved through evolution. This approach will be applicable to the isolation of other beta-actin homologues in the investigation of phylogenetic comparisons of fish species, along with a possible application to cloning strategy in other conserved genes. PMID- 10811956 TI - An Eel Cytochrome P450 1A Gene Having No XRE Sequences in Its 5' Upstream Region of 1600 bp. AB - The gene family cytochrome P450 (CYP) 1A is ubiquitous among animals, and CYP1A1 genes have been identified in teleost and mammals. We isolated another CYP1A gene, which is inducibly expressed by exposure to 3-methylcholanthrene, from the genomic library of the eel (Anguilla japonica). The genomic clone obtained, approximately 17,500 bp in length, contained the structural gene of the CYP1A and a 5' upstream region of about 1600 bp. Sequence analysis of the 5' upstream region and the first and second exons revealed that the initiation codon was in the second exon, as in the CYP1A genes reported for mammals and teleosts. CAAT and TATA boxes were found 51 and 29 bp upstream from the transcriptional start site, respectively. Unlike the CYP1A1 genes of eel and other animals, we found no xenobiotic responsive element (XRE) core sequences in the 5' upstream region studied (1631 bp) or in the first intron, whereas three peculiar regions, each composed of multiple repeats of the trinucleotide TAA, were found between 824 and 1356 bp upstream from the transcriptional start site. Absence of XRE core sequences and presence of the multiple repeats in the 5' upstream region sequenced may suggest that the expression mechanism of the eel CYP1A gene is somewhat different from the mechanisms of other reported CYP1A genes. PMID- 10811957 TI - Cloning and Expression of an Elongation Factor-1alpha in Sea Bream (Sparus aurata) Larvae and Adult Tissue. AB - A clone encoding the polypeptide elongation factor EF-1alpha was isolated from a complementary DNA library prepared from sea bream (Spartus aurata) larvae 1 to 10 days after hatching. The deduced amino acid sequence is between 82% and 95% similar to EF-1alpha in other animal species. EF-1alpha messenger RNA is present at low abundance in sea bream embryos prior to gastrulation, but at around 15 hours postfertilization, there is a 10-fold increase in transcript levels. This increase presumably reflects midblastula transition in this species. In adult sea bream, EF-1alpha appeared to have a relatively uniform distribution across all the tissues analyzed. PMID- 10811958 TI - Genetic Diversity and Species-Diagnostic Markers of Mud Crabs (Genus Scylla) in Eastern Thailand Determined by RAPD Analysis. AB - Genetic diversity of three mud crab species, Scylla serrata (Forskal), S. oceanica (Dana), and S. tranquebarica (Fabricius), collected from two locations in eastern Thailand (Chanthaburi and Trat) was examined by randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Ninety-one reproducible RAPD fragments, generated by UBC456, UBC457, and YNZ22, were polymorphic. The percentage of polymorphic bands within populations ranged from 47.92% to 77.59%. Species-specific RAPD markers were also observed and used to construct a molecular diagnostic key in these taxa. Large genetic differences between species were found (D(ij) = 0.425 to 0.751), whereas those between populations within each species were much lower (D(ij) = 0.171 to 0.199). The neighbor-joining tree based on genetic distances among pairs of individuals indicated three distinct groups, corresponding to S. serrata, S. oceanica, and S. tranquebarica. No genotypes were shared among these three species. This suggests the absence of genetic exchanges between sympatric mud crab species in eastern Thailand. Therefore, mud crabs in this area should be recognized as three different species rather than a single panmictic species exhibiting different morphs. PMID- 10811959 TI - Occurrence of Marine Birnavirus Through the Year in Coastal Seawater in the Uwa Sea. AB - This study aims to determine the seasonal occurrence of marine birnavirus (MABV) at a coastal site in the Uwa Sea, Japan, in 1997 and 1998. To detect MABV from seawater, a simple method was developed for concentrating MABV by dialysis and ethanol precipitation. The concentrated virus was used for polymerase chain reaction (PCR), enzyme-linked immunosorbent assay, and virus isolation. Viral genome was detected through the experimental period. The amount of PCR product varied; it was small in summer, but increased from fall to winter. Viral protein was also detected, and the amount in the January sample was equivalent to approximately 10(2) TCID(50) (50% tissue culture infectious dose) of the virus. However, infectious viruses were not isolated. This suggested that MABV was released from hosts to environmental seawater in winter and possibly degraded after release. PMID- 10811960 TI - Metallothionein Isoforms in Mytilus edulis (Mollusca, Bivalvia): Complementary DNA Characterization and Quantification of Expression in Different Organs after Exposure to Cadmium, Zinc, and Copper. AB - Reverse transcriptase-polymerase chain reaction has been used to isolate one metallothionein isoform (MT-20) complementary DNA from RNA extracted from mussel gill. Another cDNA, isolated by screening a Mytilus edulis cDNA mantle library using the first cDNA as probe, codes for the MT-10 IV isoform. Northern blot analysis using these cDNAs revealed different expression of these isoforms. Induction with CdC1(2) caused high levels of both MT messenger RNAs, especially the MT-20, which was induced by cadmium salt but not by zinc and copper salts. An induction of MT-10 was detected with ZnCl(2). These results show that genes encoding distinct MT isoforms are differentially regulated by heavy metals. PMID- 10811961 TI - Gene Structure of the U2 snRNP-Specific A' Protein Gene from Salmo salar: Alternative Transcripts Observed. AB - The genomic sequence of the Atlantic salmon (Salmo salar) U2 snRNP-specific A' protein gene (U2A') was identified on a cosmid clone, and the exon/intron organization of a U2A' gene is described for the first time. The cosmid sequence includes 8 exons from which the 227 amino-terminal amino acids, of a total of 339 amino acids, are deduced. The cosmid cloning site was found in the 8th intron, which also contains a BglII minisatellite repeat region of unknown length. The 3' end of the gene was determined from a complementary DNA sequence. In the 234 amino-terminal amino acids the sequence identity is 84% to human and 49% to Arabidopsis thaliana. The phases of introns 1 to 8 are 1, 2, 0, 2, 0, 2, 0, and 1, respectively. The salmon U2A' gene was found by reverse transcription polymerase chain reaction to be expressed in all 10 tissues tested, and in addition to the expected fragment of 695 bp, a smaller fragment of 615 bp was amplified in all tissues. Nucleotide sequencing of the fragments obtained showed that the smaller transcript lacks exon 6 and this results in frameshift of exon 7 and a truncated deduced polypeptide. This result may be explained by alternative splicing of the salmon U2A' gene. Different U2A' transcripts were found to exist also in human tissue. PMID- 10811962 TI - Molecular properties of sodium/dicarboxylate cotransporters. PMID- 10811963 TI - Diffusion of small solutes in the lateral intercellular spaces of MDCK cell epithelium grown on permeable supports. AB - The diffusion coefficients of four solutes ranging in molecular weight from 238 to 10,000 in the lateral intercellular spaces (LIS) of cultured kidney cells (MDCK) grown on permeable supports were determined from the spread of fluorescence produced after the release of caged compounds by a pulse from a UV laser. Two types of experiments were performed: measurement of the rate of change of fluorescence after releasing a caged fluorophore, and measurement of the change in fluorescence of a relatively static fluorescent dye produced by the diffusion of an uncaged ligand for the dye. Fluorescence intensity was determined by photon-counting the outputs of a multichannel photomultiplier tube. Diffusion coefficients were determined in free solution as well as in the LIS of MDCK cells grown on permeable supports and the hindrance factor, theta, determined from the ratio of the free solution diffusivity to that in the LIS. The hindrance factors for 3000-MW dextran, 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS, MW 524) and N 2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES, MW 238) were not significantly different from 1. The diffusion of 10,000-MW dextran was substantially reduced in the LIS with a theta of 5.6 +/- 0.3. Enzymatic digestion by neuraminidase of the sialic acid residues of the glycosylation groups in the LIS increased the diffusivity of the 10,000-MW dextran 1.8-fold indicating hindrance by the glycocalyx. We conclude that small solutes, such as Na(+) and Cl(-), would not be significantly restricted in their diffusion in the LIS and that solute concentration gradients could not develop along the LIS under physiologic conditions. PMID- 10811964 TI - Voltage gating of gap junctions in cochlear supporting cells: evidence for nonhomotypic channels. AB - The organ of Corti has been found to have multiple gap junction subunits, connexins, which are localized solely in nonsensory supporting cells. Connexin mutations can induce sensorineural deafness. However, the characteristics and functions of inner ear gap junctions are not well known. In the present study, the voltage-dependence of gap junctional conductance (G(j)) in cochlear supporting cells was examined by the double voltage clamp technique. Multiple types of asymmetric voltage dependencies were found for both nonjunctional membrane voltage (V(m)) and transjunctional (V(j)) voltage. Responses for each type of voltage dependence were categorized into four groups. The first two groups showed rectification that was polarity dependent. The third group exhibited rectification with either voltage polarity, i.e., these cells possessed a bell-shaped G(j)-V(j) or G(j)-V(m) function. The rectification due to V(j) had fast and slow components. On the other hand, V(m)-dependent gating was fast (<5 msec), but stable. Finally, a group was found that evidenced no voltage dependence, although the absence of V(j) dependence did not preclude V(m) dependence and vice versa. In fact, for all groups V(j) sensitivity could be independent of V(m) sensitivity. The data show that most gap junctional channels in the inner ear have asymmetric voltage gating, which is indicative of heterogeneous coupling and may result from heterotypic channels or possibly heteromeric configurations. This heterogeneous coupling implies that single connexin gene mutations may affect the normal physiological function of gap junctions that are not limited to homotypic configurations. PMID- 10811965 TI - Large conductance Ca(2+)-activated K(+) channels in human meningioma cells. AB - Cells from ten human meningiomas were electrophysiologically characterized in both living tissue slices and primary cultures. In whole cells, depolarization to voltages higher than +80 mV evoked a large K(+) outward current, which could be blocked by iberiotoxin (100 nm) and TEA (half blocking concentration IC(50) = 5.3 mm). Raising the internal Ca(2+) from 10 nm to 2 mm shifted the voltage of half maximum activation (V(1/2)) of the K(+) current from +106 to +4 mV. Respective inside-out patch recordings showed a voltage- and Ca(2+)-activated (BK(Ca)) K(+) channel with a conductance of 296 pS (130 mm K(+) at both sides of the patch). V(1/2) of single-channel currents was +6, -12, -46, and -68 mV in the presence of 1, 10, 100, and 1000 microm Ca(2+), respectively, at the internal face of the patch. In cell-attached patches the open probability (P(o)) of BK(Ca) channels was nearly zero at potentials below +80 mV, matching the activation threshold for whole-cell K(+) currents with 10 nm Ca(2+) in the pipette. Application of 20 microm cytochalasin D increased P(o) of BK(Ca) channels in cell-attached patches within minutes. These data suggest that the activation of BK(Ca) channels in meningioma cells does not only depend on voltage and internal Ca(2+) but is also controlled by the cytoskeleton. PMID- 10811966 TI - Direct comparison of NPPB and DPC as probes of CFTR expressed in Xenopus oocytes. AB - Blockers of CFTR with well-characterized kinetics and mechanism of action will be useful as probes of pore structure. We have studied the mechanism of block of CFTR by the arylaminobenzoates NPPB and DPC. Block of macroscopic currents by NPPB and DPC exhibited similar voltage-dependence, suggestive of an overlapping binding region. Kinetic analysis of single-channel currents in the presence of NPPB indicate drug-induced closed time constants averaging 2.2 msec at -100 mV. The affinity for NPPB calculated from single-channel block, K(D) = 35 microm, exceeds that for other arylaminobenzoates studied thus far. These drugs do not affect the rate of activation of wild-type (WT) channels expressed in oocytes, consistent with a simple mechanism of block by pore occlusion, and appear to have a single binding site in the pore. Block by NPPB and DPC were affected by pore domain mutations in different ways. In contrast to its effects on block by DPC, mutation T1134F-CFTR decreased the affinity and reduced the voltage-dependence for block by NPPB. We also show that the alteration of macroscopic block by NPPB and DPC upon changes in bath pH is due to both direct effects (i.e., alteration of voltage-dependence) and indirect effects (alteration of cytoplasmic drug loading). These results indicate that both NPPB and DPC block CFTR by entering the pore from the cytoplasmic side and that the structural requirements for binding are not the same, although the binding regions within the pore are similar. The two drugs may be useful as probes for overlapping regions in the pore. PMID- 10811967 TI - Influence of membrane physical state on the lysosomal proton permeability. AB - Influence of membrane physical state on the proton permeability of isolated lysosomes was assessed by measuring the membrane potential with 3,3' dipropylthiadicarbocyanine iodide and monitoring their proton leakage with p nitrophenol. Changes in the membrane order were examined by the steady-state fluorescence anisotropy of 1, 6-diphenyl-1,3,5-hexatriene. Both the membrane potential and proton leakage increased with fluidizing the lysosomal membranes by benzyl alcohol and decreased with rigidifying the membranes by cholesteryl hemisuccinate. The proton permeability increased to the maximum of 42% by the benzyl alcohol treatment and decreased to the minimum of 38.1% by the cholesteryl hemisuccinate treatment. Treating the lysosomes with protonophore CCCP increased the proton permeability by 58%. The effects of the membrane fluidization and rigidification can be reversed by rigidifying the fluidized membranes and fluidizing the rigidified membranes, respectively. The results indicate that the proton permeability of lysosomes increased and decreased with increasing and decreasing their membrane fluidity, respectively. Moreover, the lysosomal proton permeability did not alter further if the changes, either an increase or a decrease, in the fluidity exceeded some amount. The results suggest that the proton permeability of lysosomes can be modulated finitely by the alterations in their membrane physical state. PMID- 10811968 TI - Involvement of protein tyrosine kinase in osmoregulation of Na(+) transport and membrane capacitance in renal A6 cells. AB - Renal A6 cells have been reported in which hyposmolality stimulates Na(+) transport by increasing the number of conducting amiloride-sensitive 4-pS Na(+) channels at the apical membrane. To study a possible role of protein tyrosine kinase (PTK) in the hyposmolality-induced signaling, we investigated effects of PTK inhibitors on the hyposmolality-induced Na(+) transport in A6 cells. Tyrphostin A23 (a PTK inhibitor) blocked the stimulatory action of hyposmolality on a number of the conducting Na(+) channels. Tyrphostin A23 also abolished macroscopic Na(+) currents (amiloride-sensitive short-circuit current, I(Na)) by decreasing the elevating rate of the hyposmolality-increased I(Na). Genistein (another type of PTK inhibitor) also showed an effect similar to tyrphostin A23. Brefeldin A (BFA), which is an inhibitor of intracellular translocation of protein, blocked the action of hyposmolality on I(Na) by diminishing the elevating rate of the hyposmolality-increased I(Na), mimicking the inhibitory action of PTK inhibitor. Further, hyposmolality increased the activity of PTK. These observations suggest that hyposmolality would stimulate Na(+) transport by translocating the Na(+) channel protein (or regulatory protein) to the apical membrane via a PTK-dependent pathway. Further, hyposmolality also caused an increase in the plasma (apical) membrane capacitance, which was remarkably blocked by treatment with tyrphostin A23 or BFA. These observations also suggest that a PTK-dependent pathway would be involved in the hyposmolality-stimulated membrane fusion in A6 cells. PMID- 10811969 TI - Lipid phase fatty acid flip-flop, is it fast enough for cellular transport? AB - The mechanism by which fatty acids are transported across cell membranes is controversial. The essence of the controversy is whether transport requires membrane protein mediation or whether the membrane's lipid phase provides a pathway so rapid that a protein is not needed. This review focuses on the mechanisms of fatty acid transport across lipid bilayer membranes. These results for lipid membranes are used to help evaluate transport across cell membranes. Within the context of this analysis, a lipid phase mediated process is consistent with results for the transport of fatty acids across erythrocytes but provides a less adequate explanation for fatty acid transport across more complex cells. PMID- 10811970 TI - Effects of Na(+) on the predominant K(+) channel in the tonoplast of Chara: decrease of conductance by blocks in 100 nanosecond range and induction of oligo- or poly-subconductance gating modes. AB - We present three mechanisms by which Na(+) inhibits the open channel currents of the predominant K(+) channel in the tonoplast of Chara corallina: (i) Fast block, i.e., short (100 ns range) interruptions of the open channel current which are determined by open channel noise analysis, (ii): Oligo-subconductance mode, i.e., a gating mode which occurs preferentially in the presence of Na(+); this mode comprises a discrete number (here 3) of open states with smaller conductances than normal, and (iii): Polysubconductance mode, i.e., a gating mode with a nondiscrete, large number (>30) of states with smaller conductances than the main open channel conductance. This novel mode has also been observed only in the presence of Na(+). PMID- 10811971 TI - Permeability of boric acid across lipid bilayers and factors affecting it. AB - Boron enters plant roots as undissociated boric acid (H(3)BO(3)). Significant differences in B uptake are frequently observed even when plants are grown under identical conditions. It has been theorized that these differences reflect species differences in permeability coefficient of H(3)BO(3) across plasma membrane. The permeability coefficient of boric acid however, has not been experimentally determined across any artificial or plant membrane. In the experiments described here the permeability coefficient of boric acid in liposomes made of phosphatidylcholine was 4.9x10(-6) cm sec(-1), which is in good agreement with the theoretical value. The permeability coefficient varied from 7x10(-6) to 9.5x10(-9) cm sec(-1) with changes in sterols (cholesterol), the type of phospholipid head group, the length of the fatty acyl chain, and the pH of the medium. In this study we also used Arabidopsis thaliana mutants which differ in lipid composition to study the effect of lipid composition on B uptake. The chs1 1 mutant which has lower proportion of sterols shows 30% higher B uptake compared with the wild type, while the act1-1 mutant which has an increased percentage of longer fatty acids, exhibited 35% lower uptake than the wild type. Lipid composition changes in each of the remaining mutants influenced B uptake to various extents. These data suggest that lipid composition of the plasma membrane can affect total B uptake. PMID- 10811972 TI - Effect of ferriprotoporphyrin IX and non-heme iron on the Ca(2+) pump of intact human red cells. AB - Previous studies have shown that ferriprotoporphyrin IX (FP) and non-heme iron have a marked inhibitory effect on the Ca(2+)-Mg(2+)-ATPase activity of isolated red cell membranes, the biochemical counterpart of the plasma membrane Ca(2+) pump (PMCA). High levels of membrane-bound FP and non-heme iron have been found in abnormal red cells such as sickle cells and malaria-infected red cells, associated with a reduced life span. It was important to establish whether sublytic concentrations of FP and non-heme iron would also inhibit the PMCA in normal red cells, to assess the possible role of these agents in the altered Ca(2+) homeostasis of abnormal cells. Active Ca(2+) extrusion by the plasma membrane Ca(2+) pump was measured in intact red cells that had been briefly preloaded with Ca(2+) by means of the ionophore A23187. The FP and nonheme iron concentrations used in this study were within the range of those applied to the isolated red cell membrane preparations. The results showed that FP caused a marginal inhibition ( approximately 20%) of pump-mediated Ca(2+) extrusion and that non-heme iron induced a slight stimulation of the Ca(2+) efflux (11-20%), in contrast to the marked inhibitory effects on the Ca(2+)-Mg(2+)-ATPase of isolated membranes. Thus, FP and non-heme iron are unlikely to play a significant role in the altered Ca(2+) homeostasis of abnormal red cells. PMID- 10811973 TI - Kinetic properties of the channels formed by the bacillus thuringiensis insecticidal crystal protein Cry1C in the plasma membrane of Sf9 cells. AB - Spectrofluorimetric measurements were conducted to quantify, in real-time, membrane permeability changes resulting from the treatment of Sf9 insect cells (Spodoptera frugiperda, Lepidoptera) with different Bacillus thuringiensis Cry insecticidal proteins. Coumarin-derived CD222 and Merocyanin-540 probes were respectively used to monitor extracellular K(+) and membrane potential variations upon Sf9 cells incubation with Cry toxins. Our results establish that Cry1C induces, after a delay, the depolarization of the cell membrane and the full depletion of intracellular K(+). These changes were not observed upon Sf9 cells treated with Cry1A family toxins. Both the rate of the K(+) efflux and the delay before its onset were dependent on toxin concentration. Both parameters were sensitive to temperature but only the delay was affected by pH. Cry1C-induced K(+) efflux was inhibited by lanthanum ions in a dose-dependent manner. This study provides the first kinetic and quantitative characterization of the ion fluxes through the channels formed by a Cry toxin in the plasma membrane of a susceptible insect cell line. PMID- 10811974 TI - Very negative potential for half-inactivation of, and effects of anions on, voltage-dependent sodium currents in acutely isolated rat olfactory receptor neurons. AB - Previous measurements with CsF pipette solutions using whole-cell patch-clamp techniques in dissociated rat olfactory receptor neurons (ORNs) indicated that the sodium currents had very negative inactivation characteristics with the implication that the cell resting potential must also normally have a very negative value. This study supports the conclusions that such an effect was real and not dependent on either the nature of the pipette anions or the recording situation previously used. For all pipette solutions, sodium currents showed a threshold activation approximately -80 mV and half-maximal activation voltages approximately -55 with half-inactivation potential < or =-100 mV, without being significantly affected by the replacement of F(-) by other pipette anions (H(2)PO(-)(4) and acetate(-)) or the addition of nucleotides and glutathione (which did cause a very slight positive shift). F(-), followed by H(2)PO(-)(4) and to a much lesser extent by acetate(-), was the most favorable pipette anion for obtaining good seals and whole-cell sodium currents in these extremely small ORNs. These results implied that resting potentials, for viable responsive cells, should be more negative than about -90 mV, as supported by the observation that action potentials could only be evoked from holding potentials more negative than -90 mV. PMID- 10811976 TI - Three types of membrane excitations in the marine diatom Coscinodiscus wailesii. AB - Three types of electrical excitation have been investigated in the marine diatom Coscinodiscus wailesii. I: Depolarization-triggered, transient Cl(-) conductance, G(Cl)(t), followed by a transient, voltage-gated K(+) conductance, G(K), with an active state a and two inactive states i(1) and i(2) in series (a-i(1)-i(2)). II: Similar G(Cl)(t) as in Type-I but triggered by hyperpolarization; a subsequent increase of G(K) in this type is indicated but not analyzed in detail. III: Hyperpolarization-induced transient of a voltage-gated activity of an electrogenic pump (i(2)-a-i(2)), followed by G(Cl)(t) as in Type-II excitations. Type-III with pump gating is novel as such. G(Cl)(t) in all types seems to reflect the mechanism of InsP(-)(3) and Ca(2+)-mediated G(Cl)(t) in the action potential in Chara (Biskup et al., 1999). The nonlinear current-voltage-time relationships of Type-I and Type-III excitations have been recorded under voltage clamp using single saw-tooth command voltages (voltage range: -200 to +50 mV, typical slope: +/-1 Vs(-1)). Fits of the corresponding models to the experimental data provided numerical values of the model parameters. The statistical significance of these solutions is investigated. We suggest that the original function of electrical excitability of biological membranes is related to osmoregulation which has persisted through evolution in plants, whereas the familiar and osmotically neutral action potentials in animals have evolved later towards the novel function of rapid transmission of information over long distances. PMID- 10811975 TI - Calcium mediates the NO-induced potassium current in toad and rat olfactory receptor neurons. AB - Nitric oxide (NO) activates a K(+) current in dissociated amphibian olfactory receptor neurons. Using the patch-clamp technique in its whole-cell mode and stimulation with puffs of the NO-donor sodium nitroprusside, we further studied this effect and show that it was sensitive to the K(+)-channel blockers tetraethylammonium and iberiotoxin, indicating the activation of a Ca(2+) dependent K(+) conductance. The Ca(2+)-channel blockers nifedipine and cadmium abolished the NO-induced current, and lowering external Ca(2+) reduced it significantly. Ca(2+) imaging showed a transient fluorescence increase upon stimulation with NO, and after blockade of K(+) currents, an NO-induced inward current could be measured, suggesting that the activation of the Ca(2+)-dependent K(+) conductance is mediated by Ca(2+) influx. LY83583, a blocker of the ciliary cAMP-gated channels, did not affect the current, and experiments with focal stimulation indicated that the effect is present in the soma, therefore Ca(2+) is unlikely to enter via the transduction channels. Finally, we show that NO exerts an effect with similar characteristics on olfactory receptor neurons from the rat. These data represent the first evidence that NO activates a Ca(2+)-dependent K(+) conductance by causing a Ca(2+) influx in a sensory system, and suggest that NO signaling plays a role in the physiology of vertebrate olfactory receptor neurons. PMID- 10811978 TI - Morphogenetic and proliferative responses to heregulin of mammary epithelial cells in vitro are dependent on HER2 and HER3 and differ from the responses to HER2 homodimerisation or hepatocyte growth factor. AB - The effects of heregulin on the cell line HB2, derived from immortalised human luminal mammary epithelial cells, have been examined. HB2 cells, which normally form smooth spherical colonies in collagen gels, exhibited a striking heregulin induced morphological change to colonies projecting a large number of spiky branches. A mitogenic effect of heregulin on HB2 cells was also seen, which was more pronounced on collagen than on plastic, whereas cell motility was unaffected. HB2 cells were found to express the heregulin receptor subunits HER2 and HER3, but not HER4. Treatment of HB2 cells with heregulin also induced tyrosine phosphorylation of a band shown by immunoprecipitation to contain HER3. Using specific receptor-blocking antibodies, it was found that both the morphogenetic and proliferative responses of heregulin in HB2 cells were mediated by HER2 and HER3. To compare the effects of HER2 in heregulin signaling to heregulin-independent HER2 homodimerisation (thought to be a carcinoma-associated event), HB2 cells were transfected with the trk-neu hybrid receptor which could be induced to form homodimers by NGF. Although activated HER2 homodimers induced proliferation in the HB2 transfectants in collagen, a morphological response in collagen was not seen, suggesting that HER3 signaling is important for morphogenesis in this cell type. PMID- 10811979 TI - Inhibition of prostate specific antigen expression by genistein in prostate cancer cells. AB - Recent studies have provided convincing evidence for the role of soy-isoflavones, particularly genistein, in the inhibition of prostate cancer cell growth. Prostate specific antigen (PSA) is a biological marker used to detect and monitor the treatment of prostate cancer patients. Previous studies have documented that isoflavones can inhibit the secretion of PSA in the androgen-dependent prostate cancer cell line, LNCaP, however, the effects of genistein on androgen independent PSA expression has not been explored. In this study, we have utilized a prostate cancer cell line, VeCaP, which expresses PSA in an androgen independent manner, to determine the effects of genistein on cell proliferation and PSA expression. Here we show that genistein inhibits cell growth similarly in both the LNCaP and VeCaP cell lines, but has differential effects on PSA expression. We demonstrate using concentrations of genistein that have been detected in the serum of humans consuming a soy-rich diet, that genistein decreases PSA mRNA, protein expression and secretion. Conversely, only high concentrations of genistein inhibited PSA expression in VeCaP cells. Additionally, we have demonstrated that genistein inhibits cell proliferation independent of PSA signaling pathways, providing further evidence to support the role of genistein as a chemopreventive/therapeutic agent for prostate cancer irrespective of androgen responsiveness. PMID- 10811980 TI - Delimiting the use of comparative genomic hybridization in human myeloid neoplastic disorders. AB - Hematopoietic disorders can be used as a suitable tool of additional information on the actual resolving power of comparative genomic hybridization (CGH). Therefore, CGH examination was performed of DNA extracted from 23 acute and 15 chronic myeloproliferative disorders which had just been analyzed using classical cytogenetic techniques. In nearly all cases CGH analysis was repeated with reversely labeled probes. A Zeiss axioplan microscope was equipped with the ISIS 3 system for photometric evaluation of the CGH data. A main group was selected of 34 cases showing karyotypic mosaics when routinely diagnosed by classical cytogenetics. The grade of mosaicism was basically determined from the classical cytogenetic analysis and was additionally defined examining target anomalies by I FISH analysis in 28 of the cases. The second group included 23 cases with deletions, and in 1 case another informative genomic imbalance could be analyzed. Every target anomaly irrespective of its type could be detected in all cases with an affected cell population equalling or exceeding about 25%, but in none was it below 23%. This value was the lowest and was found in a case, with CGH-detected 20q deletion. The smallest deletions of two bands on 20q which could visually be detected by CGH were estimated in the range of 5-7 Mb. CGH was also suitable to detect imbalances which were not clearly detected by routine cytogenetics. Reverse labelling, performed in nearly all cases, confirmed the result of the original CGH analysis. These data not only document the readiness and reliability of CGH studies on human leukemia, but also further contribute to a clearer definition of the limits of the resolving power of this technique. PMID- 10811981 TI - Evidence of involvement of the PLAG1 gene in lipoblastomas. AB - Previous cytogenetic studies have demonstrated that the majority of lipoblastomas show rearrangements, in particular translocations and insertions, with breakpoints in 8q11-13. Here we present evidence for involvement of the developmentally regulated zink finger gene PLAG1 in these rearrangements. Northern blot and RT-PCR analyses revealed overexpression of PLAG1 in two lipoblastomas. Using immunohistochemistry, expression of the PLAG1 protein was also demonstrated in tissue sections from two lipoblastomas, one of which had a t(3;8)(q13.1;q12) translocation and the other a t(1;6)(q42;p22) translocation. Since no aberrant PLAG1 transcripts could be detected, it is likely that the gene may be activated by promoter swapping/substitution or alternatively by an as yet unknown mechanism. Our findings indicate that PLAG1 activation is a recurrent event in lipoblastomas and that PLAG1 is likely to be the target gene on chromosome 8 in these tumors. PMID- 10811982 TI - N-ethylmaleimide-enhanced phosphatidylserine externalization of human pancreatic cancer cells and immediate phosphatidylserine-mediated phagocytosis by macrophages. AB - The purpose of this study was to examine whether N-ethylmaleimide (NEM) can enhance phosphatidylserine (PS) externalization on the outer surface of cancer cells, and PS-externalized cancer cells can be phagocytosed immediately by macrophages. NEM could externalize PS of cancer cells in concentration- and time dependent manners. When treated with 5 mM NEM for 1 h, 80% of the carcinoma cells externalized their PS. In phagocytosis assay, these cells were engulfed immediately by macrophages before undergoing apoptosis. These results suggest that NEM can immediately externalize PS of cancer cells, leading to their recognition and phagocytosis by macrophages before undergoing apoptosis morphologically. PMID- 10811983 TI - Nuclear accumulation of p53 protein and apoptosis induced by various anticancer agents, u.v.-irradiation and heat shock in primary normal human skin fibroblasts. AB - Relationship between nuclear accumulation of p53 protein and apoptosis induced by both DNA damaging agents (ultraviolet-irradiation and anticancer agents) and heat shock in normal human skin fibroblasts was investigated. Nuclear accumulation of p53 protein and apoptosis were induced dose-dependently by u.v. -irradiation and anticancer agents. Regarding heat shock treatment, intensive nuclear accumulation of p53 protein was induced by treatment at 43 degrees C for 45 min, but, apoptotic cells were never detected. These results indicated that accumulation of p53 protein and apoptosis induced by DNA damaging agents were considered to have a close relationship and the functions of induced p53 protein are different depending on the stimulating factor. PMID- 10811984 TI - Clinical significance of S100A4 and E-cadherin-related adhesion molecules in non small cell lung cancer. AB - S100A4 has been implicated in the malignant phenotype of tumor cells, including cell motility, but the biological function is hardly known. A recent study suggests that S100A4-induced invasiveness in malignant tumor cells is partially caused by down-regulation of E-cadherin. To clarify the clinical significance of S100A4 and its association with E-cadherin-mediated cell-to-cell adhesion system, we examined their protein expressions in non-small cell lung cancer (NSCLC) specimens using immunohistochemical techniques. Expression of S100A4 was observed in 81 (60%) of 135 NSCLCs and correlated with progression of the pathological T factor (p<0.001), lymph node metastasis (p<0.005), and poor survival (p<0.05). Reduced expression of E-cadherin, alpha-catenin, and beta-catenin was observed in 64% (87 of 135), 50% (43 of 86), and 58% (50 of 86) of the specimens tested, respectively. The expression of E-cadherin closely correlated with differentiation and inversely with that of S100A4. Among these adhesion associated molecules we found that alpha-catenin appeared to reflect most strikingly the presence of lymph node metastasis and the short survival periods of NSCLC patients. Furthermore, patients who showed S100A4-positive/alpha-catenin negative expression had a significantly shorter survival than the patients with S100A4-negative/alpha-catenin-positive expression. These results indicate that S100A4, as well as alpha-catenin, plays a role in the progression and metastasis of NSCLCs and that simultaneous immunohistochemical detection of their expression may be useful to define a subpopulation of lung cancer patients with a possible poor prognosis. PMID- 10811985 TI - beta-catenin mutations are absent in hepatocellular carcinomas of SV40 T-antigen transgenic mice. AB - beta-catenin mutations have been found not only in melanoma and prostatic carcinoma but also in hepatocellular carcinomas in human, c-myc, H-ras genes transgenic mice and chemically-induced models. We investigated beta-catenin mutations in human hepatocellular carcinomas (HCCs), Hep G2 cell line and HCCs in SV40 T-antigen transgenic mice, in order to examine whether beta-catenin mutations are frequently observed in HCC in general. We found a point mutation of beta-catenin in one of nine HCCs in human and a deletion of it in Hep G2 cell line. However, we found no mutation in HCC in SV40 TG mice liver. PMID- 10811986 TI - Expression of TRAIL (Apo2L), DR4 (TRAIL receptor 1), DR5 (TRAIL receptor 2) and TRID (TRAIL receptor 3) genes in multidrug resistant human acute myeloid leukemia cell lines that overexpress MDR 1 (HL60/Tax) or MRP (HL60/AR). AB - Previously we have reported a differential expression of CD95/CD95L and Bcl-2 family of genes in multidrug resistant tumor cells. TRAIL, a member of the TNF receptor family, induces apoptosis in many tumor cells by binding to DR4 (TRAIL receptor 1) and DR5 (TRAIL receptor 2). In contrast, TRAIL-induced apoptosis is prevented by a decoy receptor (DcR1, TRID or TRAIL receptor 3). In the present study, we compared the expression of TRAIL, DR4, DR5, and TRID between a drug sensitive HL60, a myeloid leukemia cell line, and its multidrug resistant (MDR) sublines that either overexpressed MDR 1 gene (HL60/Tax) or MRP gene (HL60/AR), using RT-PCR. TRAIL mRNA was expressed in HL60 cells but was present in low levels in HL60/AR cells and was completely lacking in HL60/Tax cells. Both DR4 and DR5 were undetectable in HL60/Tax but were present at comparable levels in HL60/AR and drug sensitive HL60 cells. TRID were absent in HL60 and HL60/Tax cells, but was present in low but comparable levels in peripheral blood mononuclear cells and HL60/AR cells. These data suggest that the multidrug resistance in MDR HL60 cell lines, regardless of overexpression of MDR 1 or MRP, may be due to different mechanisms. In HL60/AR cells it appears that MDR may be due to decreased expression of TRAIL and constitutive expression of TRID, whereas in HL60/Tax cells, MDR could be due to the absence of TRAIL and/or DR4 and DR5. PMID- 10811987 TI - Diverse regulations of albumin gene expression by hepatocyte growth factor in HepG2 human hepatoma cells and primary culture of rat hepatocytes. AB - The effects of HGF on albumin gene expression in HepG2 human hepatoma cells and rat hepatocytes were investigated. HGF reduced the levels of albumin mRNA in HepG2 cells but the level was augmented in rat hepatocytes. By the transfection assay, HGF stimulated albumin promoter activity but repressed alpha-fetoprotein (AFP) enhancer activity regulating both AFP and albumin promoters in HepG2 cells. In contrast, HGF stimulated albumin promoter and AFP enhancer activities in rat hepatocytes. These results suggest that HGF elicits diverse responses of albumin gene expression in HepG2 cells and rat hepatocytes through the different biological actions on AFP enhancer in these cells. PMID- 10811988 TI - RT-PCR detection of breast cancer cells in sentinel lymph modes. AB - Sentinel lymph node (SLN) biopsy is being evaluated in breast cancer patients to improve detection of metastases and to guide therapy with minimal morbidity. The aim of this study was to increase the sensitivity of tumor cell detection in SLNs using superior reverse transcription polymerase chain reaction (RT-PCR) for carcinoembryonic antigen (CEA) and mammaglobin (MMG) analysis rather than current methods which fail to identify clinically relevant disease in many patients. In seventy stage I and II breast cancer patients dye-guided lymph node mapping was performed and the SLNs were divided alternately for RT-PCR or hematoxylin and eosin staining (H&E). RT-PCR and H&E diagnosis of SLNs were compared. SLNs were identified in 66/70 (94.3%) patients. Seventeen patients (26. 2%) had histological metastasis in SLNs. CEA was expressed in 25.0% of 48 patients with H&E negative SLNs, and MMG was expressed in 20. 8%. SLNs could predict axillary lymph node status with 95.4% accuracy and 6.3% false negative rate by H&E. Moreover, RT-PCR improved these to 98.5% and 2.8%, respectively. SLN diagnosis using RT-PCR is a powerful and sensitive method, which increases the accuracy of clinical staging and may provide more informed choices for appropriate therapeutic management of breast cancer patients. PMID- 10811989 TI - Tumor regression by inductive hyperthermia combined with hepatic embolization using dextran magnetite-incorporated microspheres in rats. AB - We formulated a novel preparation of microspheres incorporating dextran magnetite complex (DM-MS), which enable magnetic field-induced heating for the induction of hyperthermia. Using a 500 kHz magnetic field combined with arteriolar embolization of DM-MS, inductive hyperthermia was performed for the treatment of liver tumors in rats. Three days after treatment, the percentage increase of tumor volume in the embolo-hyperthermia, embolization alone, and control groups were 28%, 124%, and 385%, respectively, with the embolo-hyperthermia group differing significantly from the remaining groups. This study demonstrated the feasibility of combining embolization of DM-MS with hyperthermia as an antitumor therapy. PMID- 10811990 TI - The expression of MHC class I, TAP1/2, and LMP2/7 gene in human gastric cancer cell lines. AB - Intracellular antigens are presented to CD8+ T cells through major histocompatibility complex (MHC) class I molecule. For stable MHC class I expression, several molecules as well as the MHC molecule itself have to express simultaneously and function well. To determine a gene associated with MHC class I surface expression, the expressions of LMP2/7 and TAP1/2 including MHC I gene were analyzed in ten human gastric cancer cell lines, using RT-PCR and Northern blot analysis. Although LMP2, TAP1/2, and MHC class I gene expression were reduced in some cells, this was not significantly associated with MHC class I surface expression. By comparison, the expression of LMP7 was significantly reduced in three of ten cell lines, which also showed low levels of MHC class I surface expression, and increased in four of ten cell lines, which also showed high levels of MHC class I surface expression. These results suggest that the level of MHC class I surface expression is associated, in most cases, with the expression of the LMP7 gene regardless of the LMP2, TAP1, TAP2, or MHC class I genes. PMID- 10811992 TI - Prognostic significance of hTERT expression in non-small cell lung cancer. AB - To investigate the prognostic role of hTERT expression in non-small cell lung cancer (NSCLC), we examined the expression of hTERT mRNA in tumor specimens from 68 patients with NSCLC using RT-PCR. The expression of hTERT was detected in 34 (50%) of 68 cancer tissues. There were no correlations between hTERT status and any common clinical features except age. Patients with hTERT expression had shorter survival than those without hTERT expression. Multivariate analysis showed that hTERT expression was an independent negative prognostic factor. These results suggest that expression of hTERT may be an independent prognostic factor for NSCLC patients. PMID- 10811991 TI - Outcome of pancreatic cancer patients based on genetic lymph node staging. AB - We prospectively assessed the clinical value of genetic staging of lymph node metastasis in patients with pancreatic adenocarcinoma who underwent curative surgery. K-ras gene mutations were detected in the primary tumors in 18 of 25 patients with pancreatic adenocarcinoma. Among these 18 patients, mutated K-ras gene was also found in at least one lymph node in 13 patients. Of these 13 patients, seven had no evidence of histological nodal involvement and six had histological lymph node metastasis. Although there was no significant difference in overall survival rates between the pathological node-negative and -positive patients, overall survival of the five patients with nodes-negative for the mutated K-ras gene were significantly better than that of the 13 patients with genetically metastasis-positive nodes (p<0.001). Furthermore, overall survival of the six patients with genetically metastasis-positive nodes limited to peripancreatic area was significantly better than that of seven patients with genetical metastasis in lymph nodes beyond the peripancreatic areas (p=0.018). These findings suggest that detection of K-ras gene mutations in lymph nodes may be clinically useful to assess the accurate tumor staging and to stratify the patient with pancreatic adenocarcinoma who are at high or low risk for recurrence after curative surgery. PMID- 10811993 TI - Differences in protein kinase C and estrogen receptor alpha, beta expression and signaling correlate with apoptotic sensitivity of MCF-7 breast cancer cell variants. AB - Widespread use of MCF-7 human breast cancer cells as a model system for breast cancer has lead to variations in these cells between different laboratories. Although several reports have addressed these differences in terms of proliferation and estrogenic response, differences in sensitivity to apoptosis have just begun to be described. Based on the possible differences in apoptotic sensitivity that may arise due to the existence of MCF-7 cell variants, we determined the relative sensitivity of MCF-7 cell variants from three established laboratories (designated M, L and N) to known inducers of apoptosis. Consistent with our previous studies we demonstrate that differences exist among these variants in regards to tumor necrosis factor alpha (TNF)-induced cell death and inhibition of proliferation in a dose-dependent manner. To establish if the difference in apoptotic susceptibility was specific to TNF, the three MCF-7 cell variants were tested for their response to other known inducers of apoptosis: okadaic acid, staurosporine and 4-hydroxy-tamoxifen. Viability and DNA fragmentation analysis revealed a similar pattern of resistance to apoptosis by all agents in the MCF-7 M variant. The MCF-7 L variant was resistant to okadaic acid and 4-hydroxy-tamoxifen but not staurosporine. In contrast, MCF-7 N cells were sensitive to induction of apoptosis by all agents. The role of both protein kinase C (PKC) and estrogen signaling in the regulation of cell survival prompted investigation of these pathways as a mechanism for differential sensitivity of MCF-7 cell variants to apoptosis. While both estrogen receptor alpha (ERalpha) and ERbeta were expressed in MCF-7 M and N cells, the absence of ERbeta in MCF-7 L cells correlated with decreased estrogen responsiveness of the L variant. Variations in estrogenic responsiveness and PKC isoform expression may account for the enhanced susceptibility of both the L and N variants to staurosporine. PMID- 10811994 TI - Effects of the novel alphav integrin antagonist SM256 and cis-platinum on growth of murine squamous cell carcinoma PAM LY8. AB - Increased density of proliferating and migrating tumor cells and neovascular endothelial cells has been associated with tumor progression and poor prognosis in patients with squamous cell carcinoma (SCC). Tumor and neovascular endothelial cells in squamous cell carcinoma have been reported to express integrin heterodimers containing the alphav subunit, which binds to vitronectin and other extracellular matrix proteins that contain the amino acid recognition sequence Arg-Gly-Asp (RGD). In the present study, we examined the effect of the novel non peptide alphav integrin antagonist SM256 on growth of SCC line PAM LY8 in BALB/c SCID mice, and determined whether SM256 has direct inhibitory effects on growth of murine endothelial and PAM LY8 SCC cells in vitro. SM256 inhibits cell adhesion of murine cells expressing alphavbeta3 and alphavbeta5 integrins in vitro with an IC50 of 35 nM and 30 nM, respectively. Growth of Pam LY8 tumors in vivo was inhibited with 14-day continuous administration of SM256 by subcutaneous osmotic diffusion pump, during which a mean serum concentration of 56 nM was detected. While both murine aortic endothelial cells and PAM LY8 were found to express alphav integrins by fluorescence cytofluorometry, SM256 at 50 nM in MTT assay completely inhibited growth of endothelial cells, but had no significant direct effect on growth of PAM LY8 cells. We compared the effect on growth of PAM LY8 of SM256 infusion versus single agent or combination chemotherapy with a maximally tolerated dose of cis-platinum, which is used as a standard chemotherapy for SCC. When treatment was initiated at either 7 or 21 days following establishment of tumor, 14-day infusion of SM256 had an inhibitory effect on growth that was similar to that obtained with single dose cis-platinum, but no additive effect of concurrent therapy with SM256 and cis-platinum was observed. These results demonstrate the activity and feasibility of use of alphav antagonists such as SM256 for therapy of SCC. PMID- 10811995 TI - Treatment of hepatocellular cancer with the long acting somatostatin analog lanreotide in vitro and in vivo. AB - Based on the fact that somatostatin (SST) analogs have given promising results for treatment of hepatocellular cancer, we performed both in vitro and in vivo investigations to define the role of a depot formulation of the long acting SST analog lanreotide (LAN). A decrease of cells in the S-phase as compared to controls (p<0.03) followed by a significant, dose-dependent induction of apoptosis could be demonstrated in Hep G2 cells along with a dose-dependent influence of the peptide on cellular proliferation. Northern blotting demonstrated the presence of mRNA for SSTR subtypes 2, 3 and 4 in Hep G2 cells, but only slight SSTR expression in normal liver tissue. In addition, 21 untreated patients with advanced HCC not amenable to surgery were administered 30 mg of LAN by deep intramuscular injection every 14 days until documented disease progression. Fifteen of these patients also underwent scanning with commercially available 111In-DTPA-D-Phe1-Octreotide (111In-OCT) to define the in vivo expression of SSTR. No positive 111In-OCT scans were obtained, indicating the absence of relevant amounts of functional SSTR2 in HCC. One patient (5%) showed a partial response to treatment, 8 patients had stable disease (38%), while the remaining patients progressed during treatment. The median survival was 4.2 months (range 1.2-13+), and the median time to progression was 2.5 months (range, 1.5-7+). However, 4 patients (19%) had an increase in WHO performance status lasting between 2.5 and 6 months, 5 patients (24%) had an increase in body weight, while pain markedly improved in 1 additional patient (5%). In total, 5 patients (24%) had a decrease in serum-AFP levels by at least 30%. Our results clearly indicate the ability of LAN to decrease the S-phase fraction along with induction of apoptosis in Hep G2 cells in a dose-dependent manner. Our data suggest clinical potential of SST-analogs in HCC and indicate that suboptimal doses of the peptide might have been administered in our series. PMID- 10811996 TI - Microsatellite instability of dinucleotide tandem repeat sequences is higher than trinucleotide, tetranucleotide and pentanucleotide repeat sequences in prostate cancer. AB - In order to investigate whether the change in length of simple repetitive genomic sequences (microsatellite instability) is associated with prostate cancer, we analyzed 40 prostate cancer samples with 44 microsatellite loci markers on chromosomes 1, 3, 5, 6, 8, 9, 11, 13, 16, 17 and X. DNA was extracted from normal and tumor cells of 40 microdissected cancer samples, amplified by PCR and analyzed for microsatellite instability using 44 primers for dinucleotide, trinucleotide, tetranucleotide and pentanucleotide repeat sequences. The results of this study demonstrate that 45% of the prostate cancer specimens (18 out of 40) showed microsatellite instability (MSI) at a minimum of one locus using dinucleotide repeat sequences. Two out of 40 samples (5%) showed MSI at a minimum of one locus using three different trinucleotide repeat primers (AR, SR and TBP). Ten out of 40 (25%) samples showed MSI at a minimum of one locus using five different tetranucleotide repeat primers (HPRT1, HPRTII, MYCL1, RB, REN). There were no MSI observed in samples using pentanucleotide repeat sequences. There were no MSI in benign prostatic hyperplasia samples (25 samples). These experiments suggest that the microsatellite instability of dinucleotide tandem repeat sequences is much higher than trinucleotide, tetranucleotide and pentanucleotide repeat sequences in prostate cancer. The MSI with different lengths of nucleotide repeat sequences did not correlate with the stage and grades of prostate cancer. PMID- 10811997 TI - Expression of MTA1, a metastasis-associated gene with histone deacetylase activity in pancreatic cancer. AB - MTA1 has been identified as a metastasis-promoting gene, and its gene expression is correlated with serosal invasion and lymph node metastasis in the gastrointestinal and esophageal cancers. We examined MTA1 mRNA expression levels in 13 cell lines and 23 tumor tissues of pancreatic cancer by quantitative reverse transcription polymerase chain reaction. MTA1 mRNA expression was found in all the cell lines, however its levels were not correlated with the metastatic pontential of each cell line representing hematogenous distant metastasis. In the pancreatic cancer tissues, the overexpression of MTA1 mRNA (tumor/normal ratio >/=1) was observed in 12 out of 23. Pancreatic cancer tissues overexpressing MTA1 mRNA (T/N ratio >/=1) tended to have higher incidence of lymph node metastasis. These results together with previous findings in the gastrointestinal and esophageal cancers suggest that MTA1 might be involved in the progression, particularly in lymph node metastasis of pancreatic cancer. PMID- 10811998 TI - Modulation of growth of human prostate cancer cells by the N-acetylcysteine conjugate of phenethyl isothiocyanate. AB - There is growing evidence that thiol conjugates of isothiocyanates present in cruciferous vegetables are effective cancer chemopreventive and potentially active therapeutic agents. The effects of the N-acetylcysteine conjugate of phenethyl isothiocyanate (PEITC-NAC) on tumor cell growth were analyzed in human prostate cancer cell lines LNCaP, androgen-dependent, and DU-145, androgen independent. Exposure of the cells to PEITC-NAC at high concentrations caused cytolysis, while at lower concentrations PEITC-NAC mediated a dose-dependent growth modulation, with reduction of DNA synthesis and growth rate, inhibition of clonogenicity and induction of apoptosis in both types of prostate cancer cells. PEITC-NAC decreased cells in S and G2M phases of cell cycle, blocking cells entering replicating phases. In parallel, a significant enhancement of cells expressing the cell cycle regulator p21 as well as its intensity was determined using a fluorescent antibody technique. The action of PEITC-NAC was time dependent, with the magnitude of inhibition increasing to 50-65% after PEITC-NAC exposure for several days. Interaction of tumor cells with dissociation products of PEITC-NAC, PEITC and NAC, are proposed as the mechanism of growth regulation. PMID- 10811999 TI - How tumor stage affects american urologists' surveillance strategies after prostate cancer surgery. AB - The factors which influence decision-making among urologists are not well understood. We evaluated how tumor stage in patients subjected to potentially curative surgery for carcinoma of the prostate affects the self-reported follow up strategies employed by practicing United States urologists. Standardized patient profiles and a detailed questionnaire based on these profiles were mailed to 4,467 randomly selected members of the American Urological Association (AUA), comprising 3,205 US and 1,262 non-US urologists. The effect of TNM stage on the surveillance strategies chosen by respondents was analyzed by repeated-measures ANOVA. There were 1, 050 respondents who provided evaluable data of whom 760 were from the US. The three most commonly used surveillance modalities by urologists were office visit, serum PSA level, and urinalysis. Nine of the 11 most commonly requested modalities were ordered significantly (p<0.001) more frequently with increasing TNM stage. This effect persisted through 10 years of follow-up, but the differences across stage were tiny. Fifty-five percent of US respondents do not modify their strategies at all according to the patient's TNM stage. Most American AUA members performing surveillance after potentially curative radical prostatectomy for otherwise healthy patients use the same follow-up strategies irrespective of TNM stage. These data permit the rational design of a randomized clinical trial of two alternate follow-up plans. The two trial arms would employ office visits, blood tests, and urinalyses at different frequencies based on current actual practice patterns; there would be no imaging tests in either arm. PMID- 10812000 TI - Phase II study of induction chemotherapy followed by concomitant chemoradiotherapy in advanced head and neck cancer: clinical outcome, toxicity and organ/function preservation. AB - The purpose of the study was to assess response rate, clinical outcome, organ/function preservation and toxicity in head and neck cancer patients treated with induction chemotherapy followed by concomitant chemoradiotherapy and, when necessary, limited surgery. The study design was a phase II non-randomized trial in hospitalized patients setting. The treatment plan consisted of 3 cycles of induction chemotherapy with cisplatin, fluorouracil (5-FU), leucovorin and interferon alpha2b (PFL-IFN) followed by 7 cycles of 5-FU, hydroxyurea and concomitant radiation for 5 days (FHX) for a total radiation dose of 70 Gy. Surgical resection was performed, when necessary, with the intent to spare organ/function. Seventeen patients were treated at one institution. Three patients had stage III and 14 patients stage IV disease. Twelve patients were analyzed for response to PFL-IFN: 2/12 (16.7%) patients achieved a CR and 10/12 (83.3%) achieved a PR for an ORR of 100%. FHX was administered on protocol to 10 patients: 4 patients (40%) had CR, 3 (30%) had PR >/=70% for an ORR of 70%, 1 patient (10%) had SD and 2 patients (20%) had PD. As for local therapy, of the 8 eligible patients who completed chemoradiotherapy, the 3 patients with CR were submitted to random biopsies, results of which were histologically negative, 3 patients with PR >/=70% underwent conservative organ-preserving surgery, and 1 patient with PR >70% refused surgery, whereas the patient with SD underwent salvage surgery, preserving voice. Thus, organ preservation was achieved in all 8 patients at the completion of all therapy: 4 patients had no surgical procedure and 4 patients only conservative surgery. Overall, after completion of all therapy, 5/8 (62.5%) patients were rendered disease-free. The median overall survival time was 23 months, the median duration of response was 6 months and the median time to progression was 9 months. Both induction chemotherapy and concomitant chemoradiotherapy resulted in significant toxicity, which consisted mainly of mucositis and thrombocytopenia. In conclusion, PFL-IFN was very active, producing high ORRs and, followed by FHX, resulted in high overall survival rates permitting an optimal organ preservation, at the cost of a severe toxicity. PMID- 10812001 TI - Low frequency of microsatellite instability in sporadic breast cancer. AB - In some tumors, defects in mismatch repair enzymes lead to errors in the replication of simple nucleotide repeat segments. This condition is commonly known as microsatellite instability (MSI) because of the frequent mutations of microsatellite sequences. Although the MSI phenotype is well recognized in some colon, gastric, pancreatic, and endometrial cancers, reports of MSI in breast cancer are inconsistent. The purpose of this study was to determine the presence of MSI in breast cancer and to correlate its occurrence with clinicopathological parameters, including expression of estrogen and progesterone receptors. We have analyzed the status of 10 different microsatellite loci (mono and dinucleotide repeats). Mmicrosatellite size patterns and LOH were compared in 88 paired breast cancer/peripheral-blood DNA samples. Fluorescent polymerase chain reaction (PCR) for typing microsatellites coupled with DNA fragment analysis in an automated DNA sequencer was applied. Microsatellite instability in at least two microsatellite markers was observed in 6 out of 88 (7%) of the cases, all beloging to stage II or III. LOH was found in 48/88 (55%) of the cases. Five of the six cases with MSI also had LOH in other markers different from those of MSI. These MI and LOH data were analysed using a range of clinicopathological parameters, no correlation between MSI and histopathological characteristics were found. A significant correlation was observed between MSI and negative expression of both estrogen and progesterone receptors (p<0.02), indicating a possible relatioship between specific genetic changes at these microsatellite regions and hormonal deregulation in the progresion of breast cancer. PMID- 10812002 TI - TNF-alpha induces apoptosis mediated by AEBSF-sensitive serine protease(s) that may involve upstream caspase-3/CPP32 protease activation in a human gastric cancer cell line. AB - In the present study, we investigated the role of caspase-3/CPP32 and serine protease(s) in cell death induced by TNF-alpha in SNU-16 human gastric adenocarcinoma cells. Apoptosis induced in SNU-16 cells by TNF-alpha was accompanied by the activation of caspase-3/CPP32. After treatment with TNF-alpha, PKCdelta cleaved to its characteristic 40 kDa fragment in a caspase-3/CPP32 dependent manner. Incubation with z-DEVD-fmk completely abrogated TNF-alpha induced DNA fragmentation, indicating that activation of caspase-3/CPP32 was crucially involved in TNF-alpha-induced apoptosis. In addition, serine protease inhibitor, 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), clearly inhibited all the features of apoptosis including DNA fragmentation and chromatin condensation. Furthermore, in the AEBSF treated SNU-16 cells, only intact PKCdelta was detected by immunoblot analysis, suggesting that activation of caspase-3/CPP32 was blocked. Thus, the AEBSF-sensitive step may involve an upstream caspase-3/CPP32 protease activation. Taken together, these results suggest that both caspase-3/CPP32 and serine protease(s) are activated and play an important role in TNF-alpha induced apoptosis in SNU-16 cells. PMID- 10812003 TI - 1,25-dihydroxyvitamin D3 synthetic analogs inhibit spontaneous metastases in a 1,2-dimethylhydrazine-induced colon carcinogenesis model. AB - In order to substantiate the role of vitamin D applicability for the prevention of colon cancer and its spontaneous metastases, the effect of 1,25 dihydroxyvitamin D3 and its synthetic analogs, 1, 25-dihydroxy-16,23Z-diene-26,27 hexafluoro-D3 (Ro 25-5317) and 1, 25-dihydroxy-16,23E-diene-26,27-hexafluoro-19 nor-D3 (Ro 25-9022), have been evaluated in a 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis model in Sprague-Dawley rats. In animals maintained on 2.75 nmol/kg 1,25-dihydroxyvitamin D3 diet no statistical difference was seen in tumor incidence when compared with control while in animals on 3.0 nmol/kg 1,25 dihydroxyvitamin D3 diet, the incidence of tumors was significantly lower. In animals maintained on 3.0 nmol/kg Ro 25-5317 diet also no statistical difference was seen in tumor incidence compared with control while in animals on 3. 5 nmol/kg Ro 25-5317 diet the incidence of tumors was significantly lower. The incidence of tumors in the group of animals maintained on 3.0 nmol/kg and 3.5 nmol/kg Ro 25-9022 was significantly lower, at 32.1% and 27.6% respectively, compared to control. In the two groups of animals maintained on the 1,25 dihydroxyvitamin D3 diet no significant difference in the incidence of metastasis was seen. In the group of animals maintained on 3.0 nmol/kg Ro 25-5317 diet only regional metastases were seen. However, no metastases developed in the rats on 3.5 nmol/kg Ro 25-5317 diet. After administration of 3.0 nmol/kg Ro 25-9022 diet, metastases developed in a significantly less number of animals while no metastases occurred in the rats maintained on the 3.5 nmol/kg Ro 25-9022 diet. The above studies will provide a scientific basis for the progression into further clinical trials in the treatment, and/or chemoprevention of human colorectal cancer. PMID- 10812004 TI - To move or not: how a cell responds (Review). AB - The ability of a cell to migrate is dependent on the cooperative effects of many intracellular signaling events triggered through cell surface receptors (integrins), internal cellular mechanics, and the extracellular matrix. Many proteins have been identified as being involved in cell migration but the regulatory mechanisms involved in cell migration still remain unclear. This review summarizes studies involving various signal transduction pathways and their involvement in the regulation of cell migration. The focus is on cell adhesion sites (focal adhesions) and the actin cytoskeleton. PMID- 10812005 TI - The role of oncogenic kinases in human cancer (Review). AB - Tumorigenesis in humans is a multistep process, which reflects genetic alterations that lead to cell transformation and malignancy. Cellular genes that are altered are normally involved in maintaining cell homeostasis by participating in signaling pathways tightly regulated to maintain the functional integrity of the cell. When these genes are altered they escape from the regulatory control and transmit signals that lead to the progressive conversion of normal cells into cancer cells. Oncogenic signals involve activation of kinases, which can be either a primary event when they are directly mutated in a tumor cell or a secondary event as recipients and mediators of oncogenic signals. Transmembrane (e.g. EGFR, PDGFR) or cytoplasmic (Src, Abl) tyrosine kinases are found mutated in a variety of human tumors. Cytoplasmic serine threonine kinases (Raf, Akt, Tpl-2) are also mutated or activated in several types of human malignancies. Kinases transduce signals that lead to cell proliferation or inhibition of programmed cell death by activating transcription factors (e.g. AP1, NFkappaB, Myc), inhibiting pro-apoptotic molecules (e.g. Bad, Bax), or they participate in deregulating the cell cycle control. Thus, kinases play a central role in oncogenesis rendering them putative targets for anti-cancer drug design. PMID- 10812006 TI - Adenovirus-mediated wild-type p53 overexpression reverts tumourigenicity of human mesothelioma cells. AB - Pleural malignant mesothelioma (MM) shows poor survival, regardless of tumour stage at diagnosis. MM is unresponsive to present treatment regimens and new protocols are desperately needed. The localised nature, the potential accessibility, and the relative lack of distant metastases make MM a particularly attractive candidate for somatic gene therapy. A common target for cancer gene therapy is the tumour suppressor protein p53. p53 does not seem to be mutated or deleted in MM, but it can be inactivated by binding to other proteins, like mdm2 and SV40 large T antigen. We tested the effects of a replication-deficient adenoviral vector carrying wild-type p53 cDNA in human MM cells. Our results show that >95% of MM cells were efficiently infected with 25 multiplicity of infection (MOI) of vector. Wild-type p53 was effectively expressed resulting in >80% inhibition of proliferation in MM cells. AdCMV.p53 infection induced apoptosis while controls did not show any evident morphological alterations. Ex vivo p53 gene transfer experiments inhibited tumourigenesis in nude mice. In vivo, direct intratumour injection of AdCMV.p53 arrested tumour growth and prolonged survival of treated mice. These results indicate that p53-gene therapy should be strongly exploited for clinical trials in MM patients. PMID- 10812007 TI - Role of p53 in cellular response to anticancer nucleoside analog-induced DNA damage. AB - Anticancer nucleoside analogs (e.g., ara-C, gemcitabine, fludarabine) induce apoptosis by incorporation into DNA. Removal of incorporated analogs from DNA by 3'-5' exonucleases is presumably a mechanism of drug resistance. Based on our previous observation that the 3'-5' exonuclease activity of wild-type (wt) p53 protein is able to preferentially remove mismatched nucleotides from DNA, in the present study we further investigated the ability of p53 to recognize and remove incorporated therapeutic analogs from DNA and its role in analog-induced apoptosis. We demonstrated that although the 3'-5' exonuclease of wt p53 protein was able to bind and excise the nucleoside analog residues from DNA in vitro, removal of the drug molecules from cellular DNA was slow in whole cells with wt p53 cells, and not detectable in mutant p53 cells. Furthermore, the wt p53 were more sensitive to the cytotoxic effect of the drugs compared to the p53-null or mutant cells. Incubation of ML-1 cells (wt p53) with gemcitabine caused an accumulation of p53 protein in their nuclei and preferentially induced apoptosis in the p53-positive cells, whereas the p53-negative cells remained intact. Transfection of p53-null cells with wt p53 expression vector enhanced the sensitivity of the cells to gemcitabine. Gel mobility shift assay using synthetic DNA containing gemcitabine as the probe suggests that p53 protein is likely to participate in the binding of the analog-containing DNA. Our study suggests that recognition of the incorporated nucleoside analogs in DNA by wt p53 did not confer resistance to the drugs, but it facilitated the apoptotic cell death process. PMID- 10812008 TI - H-ras-1 point mutation in malignant peripheral nerve sheath tumors: polymerase chain reaction restriction fragment length polymorphism analysis and direct sequencing from paraffin-embedded tissues. AB - It has been shown that the NF1 (neurofibromatosis type 1) gene encodes a tumor suppressor which inactivates ras proteins. Among malignant mesenchymal tumors, H ras-1 mutations have been found in malignant fibrous histiocytoma, leiomyosarcoma and embryonal rhabdomyosarcoma. However, studies on H-ras-1 mutation of many cases of malignant peripheral nerve sheath tumors (MPNST) have not been documented. Therefore, we investigated H-ras-1 mutations of MPNST. In 45 cases of MPNSTs of our files, DNA was extracted from the formalin-fixed paraffin-embedded tissue, and the mutations of the H-ras-1 gene were detected by using PCR-RFLP (polymerase chain reaction- restriction fragment length polymorphisms) method and direct sequencing. We found two cases with H-ras-1 point mutation in MPNST for the first time. Both cases showed the same mutation in codon 13.1 [GGT(Gly) to AGT(Ser) transition]. Interestingly, both cases were associated with NF1. It is possibile that the mutation of the H-ras-1 gene occurred after the mutation of the NF1 gene in the MPNST. PMID- 10812010 TI - Internally shortened menin protein as a consequence of alternative RNA splicing due to a germline deletion in the multiple endocrine neoplasia type 1 gene. AB - Multiple endocrine neoplasia type 1 (MEN 1) is an autosomal dominantly inherited cancer syndrome (OMIM 131100), with tumours in several endocrine glands. In 1997 the responsible tumour suppressor gene was identified and recently it was shown that menin, its encoded protein, represses JunD-activated gene expression. Although many MEN 1 patients have been investigated both clinically and genetically, no genotype-phenotype correlation has been found yet. The vast majority of MEN1 gene mutations involve point mutations. We describe a patient in whom a 26 base pair deletion in the MEN1 gene, comprising part of exon 3 and part of intron 3, causes activation of a cryptic donor splice site at the beginning of exon 3. This germline mutation results in an in frame deletion of 105 nucleotides in MEN1 gene mRNA, i.e. an internal deletion of 35 amino acids in the menin protein. Since the deleted region of menin has been implicated in binding to JunD, this may explain the tumourigenic effect of this mutation. The knowledge of this MEN1 gene germline defect, may be used for presymptomatic identification of MEN 1 disease gene-carriers among family-members of this proband. This enables early detection of tumour development, timely treatment and genetic counseling. PMID- 10812009 TI - Technetium tin colloid test detecting symptomless dysphagia and ACE inhibitor prevented occurrence of aspiration pneumonia. AB - Symptomless dysphagia and swallowing disorders play a very important role in the pathogenesis of aspiration pneumonia. A videofluoroscopic examination and a simple two-step swallowing provocation test (STS-SPT) could be useful for detection of swallowing disorders in elderly patients with stroke, however, there is no report on such a test for detection of symptomless dysphagia. We administered 1 ml Technetium Tin Colloid (99mTC) to the patient during sleep via a nasal catheter placed in the mouth. At 09:00 h the next day, symptomless dysphagia was checked for by imaging. Improvement of the symptomless dysphagia was observed, and thus we could prevent the occurrence of aspiration pneumonia. The 99mTC test was particularly useful in detecting symptomless dysphagia in elderly patients with stroke. PMID- 10812011 TI - Salmonella typhimurium-induced reactivation of latent HIV-1 in promonocytic U1 cells is inhibited by trovafloxacin. AB - We have previously reported that virulent Salmonella typhimurium induces replication of latent HIV-1 in U1 cells, via activation of tumor necrosis factor alpha (TNF-alpha) production. In the present study, we show that Trovafloxacin, a new quinolone antibiotic, inhibits S. typhimurium-induced TNF-alpha production and HIV-1 replication. In addition, Trovafloxacin inhibits TNF-alpha-induced reactivation of latent HIV-1 in U1 cells. The concentrations of Trovafloxacin that inhibited HIV-1 replication are comparable to the plasma and tissue levels achieved by therapeutic dosage used in the treatment of bacterial infections. Therefore, Trovafloxacin is a potential candidate for adjunct therapy in HIV-1 infection. PMID- 10812012 TI - Mitochondrial and microsomal peripheral benzodiazepine receptors in human ovarian cancer xenografts. AB - To investigate whether the density of peripheral benzodiazepine receptors (PBR) in human ovarian tumors is related to the degree of histological differentiation and possibly elucidate their pathophysiology, PBR were measured in mitochondrial (m) and microsomal (p) fractions isolated from six different human ovarian carcinomas heterotransplanted into nude mice. A specific ligand PK11195 for PBR was employed and the density of binding sites and binding affinity (KD) were computed from Scatchard analysis. The PBR density in m-fractions was 3- to 4-fold higher than in p-fractions from all tumors. PBR density in both m- and p fractions was highest in mucinous tumors with mid-high degree of differentiation. The density in serous tumor with mid-high differentiation was significantly lower than the mucinous tumor, but higher than the serous tumor with low degree of differentiation (OVCAR-3) in both m- and p-fractions. However, the PBR density in the undifferentiated tumor (IGROV1) was higher than in OVCAR-3. The KD values for PBR were very low ranging from 5.8 to 14.0 nM in all preparations. The KD values for p-fractions were generally lower than m-fractions and highly significant differences were observed in three of the six tumors. These data suggest two separate classes of PBR pertaining to m- and p-fractions and indicate that there is no clear relationship between PBR density and degree of differentiation. PMID- 10812013 TI - Altered metabolic priming by D-glucose in pancreatic islets from Goto-Kakizaki rats. AB - The possible priming by D-glucose of metabolic events in islets from control rats and Goto-Kakizaki rats (GK rats) was investigated by first incubating the islets for 120 min either in the absence of any exogenous nutrient or presence of 16.7 mM D-glucose. The islets were then incubated for a second period of 120 min either at 2.8 mM or 16. 7 mM D-glucose, the hexose being now mixed with tracer amounts of D-[U-14C]glucose and D-[5-3H]glucose. In islets from control rats first incubated in the absence of exogenous nutrient the hierarchy in the 16.7 mM/2.8 mM ratio for metabolic variables was as follows: D-[U-14C]glucose oxidation > D-[5-3H]glucose utilization and D-[U-14C]glucose conversion to amino acids > D-[U-14C]glucose conversion to acidic metabolites. When the islets from control rats were first incubated in the presence of 16.7 mM D-glucose, the preferential stimulation of mitochondrial oxidative events at high hexose concentration, as documented by the increase in the paired ratio between D-[U 14C]glucose oxidation and D-[5-3H]glucose utilization, was further enhanced. The 16.7 mM/2.8 mM ratio for the conversion of D-[U-14C]glucose to amino acids, relative to that for D-[U-14C]glucose conversion to acidic metabolites, was much lower, however, after a first incubation in the presence of D-glucose, rather than in its absence, probably as a result of the progressive exhaustion of endogenous amino acids considered as transamination partners. The major differences between these results and those obtained in islets from GK rats consisted, in the latter animals, in i) higher absolute values for all metabolic fluxes, ii) lower 16.7 mM/ 2.8 mM ratios, iii) lower paired ratio between D-[U 14C]glucose oxidation and D-[5-3H]glucose utilization, and iv) absence of a priming effect of D-glucose (16.7 mM) upon such a paired ratio in the islets incubated at 16.7 mM D-glucose during the second incubation. Taken as a whole, these observations confirm that the preferential stimulation of mitochondrial oxidative events, in response to a rise in D-glucose concentration, is impaired in islets from GK rats and extend this knowledge to the priming action of D glucose, in high concentration, on the catabolism of the hexose during a subsequent incubation. PMID- 10812014 TI - MDM2 expression is associated with progress of disease and WAF1 expression in resected lung cancer. AB - Although MDM2, p21/WAF1, and p53 are considered as regulating each other based on in vitro studies, the relation in human lung cancer is not fully understood. The expressions of these proteins were examined immunohistochemically in 112 resected non-small cell lung cancer specimens and the correlation between them were analyzed. MDM2 was expressed in 45% of all lung cancers. In advanced stage, MDM2 positive cases were observed more frequently than in early stage, showing significant difference. No significant difference was observed in the prognosis of the patients regardless of the expression of any protein. Although no correlation was observed between MDM2 expression and p53 expression, or between p21/WAF1 expression and p53 expression, MDM2 expression was strongly related with p21/WAF1 expression. Therefore, MDM2 expression may relate to the progress of the stage of lung cancer, and MDM2 expression and p21/WAF1 expression may be associated not through the p53-related pathway. PMID- 10812015 TI - Influence of different forms of fluid shear stress on vascular endothelial TGF beta1 mRNA expression. AB - This study investigated the effects of long-term exposure of steady (19 dyne/cm2), 1-Hz non-reversing pulsatile (19+/-6 dyne/cm2) and 1-Hz purely oscillatory (0+/-19 dyne/cm2) shear stress on endothelial transforming growth factor-beta1 (TGF-beta1) mRNA expression. Cultured bovine aortic endothelial cells (BAECs) were systematically exposed to the three flow conditions for periods of 2, 6, 12 and 24 h, and relative differences in TGF-beta1 mRNA levels were measured by semi-quantitative RT-PCR. In response to steady shear stress, TGF-beta1 mRNA levels normalized to no-flow controls were 1.24, 1.42, 1.30 and 1.47 at the 2-, 6-, 12- and 24-h time points, respectively. In response to non reversing pulsatile flow, these levels were 1.49, 1.64, 1.64 and 1.73, while the respective transcript levels for oscillatory flow were 1.33, 1.12, 1.12 and 1. 93. These results indicate that BAEC TGF-beta1 mRNA was up-regulated with the kinetics of the up-regulation faster for steady and non-reversing pulsatile flow than for oscillatory flow. Given the preferential localization of early atherosclerotic lesions in arterial regions exposed to low and/or oscillatory shear stress and the implication of TGF-beta1 as an athero-protective gene, these results are consistent with the notion that regions transiently exposed to oscillatory flow may be particularly prone to atherosclerosis. PMID- 10812016 TI - Effects of succinate dimethyl ester on the metabolic and hormonal response to exercise in fed and starved rats. AB - This study aims at investigating the possible beneficial effect of succinic acid dimethyl ester (SAD), injected intraperitoneally (5.0 micromol/g body wt.), upon the metabolic and hormonal response to a 60 min exercise in both fed and overnight starved rats. In fed rats, the injection of SAD minimized the fall in plasma D-glucose concentration, and the increase in plasma lactate, beta hydroxybutyrate, free fatty acid and glycerol concentrations, otherwise provoked by exercise. SAD, however, failed to prevent the decrease in plasma insulin concentration and liver glycogen content caused by exercise. Starved rats displayed lower plasma D-glucose and insulin concentrations and higher plasma beta-hydroxybutyrate and free fatty acid concentrations than fed rats. The body weight, liver weight and paraovarian fat weight, as well as the glycogen content of both liver and heart were also decreased in the starved rats. In the latter animals, the injection of SAD again opposed the exercise-induced increase in plasma beta-hydroxybutyrate, free fatty acid and glycerol concentrations, and again failed to prevent the more modest decreases in plasma insulin concentration and liver glycogen content caused by exercise in the starved, as distinct from fed rats. These findings suggest that, independently of any obvious change in plasma insulin concentration, SAD minimizes the exercise-induced mobilization and enhanced utilization of endogenous nutrients, especially fatty acids and glycerol produced by hydrolysis of triglycerides in adipose tissue, presumably through its capacity to act as an oxidizable nutrient in various cell types and as a gluconeogenic precursor in hepatocytes. PMID- 10812017 TI - Influence of serum from healthy or breast tumor-bearing women on the growth of MCF-7 human breast cancer cells. AB - Sera from women healthy (HW) or with breast (BCW), ovarian or endometrial cancer, were added (10%) to the culture media of MCF-7 cells and cell proliferation assessed 4 days later to verify: a) whether sera from BCW, obtained before or 8 days after tumor ablaction, influence the proliferation of these cells, b) whether the effects of serum from BCW are specific for mammary tumor cells. Sera from BCW, but not sera from women with ovarian or endometrial cancer, increased MCF-7 cell proliferation in comparison with sera from HW. After surgical ablation of the breast tumors, serum's ability to increase MCF-7 cell proliferation decreased significantly. These effects cannot be explained by differences on serum levels of estradiol or melatonin. These results suggest the presence of growth-promoting substances of possible tumoral origin in serum of BCW, a fact that should be considered as support for the surgical treatment of tumor masses. PMID- 10812018 TI - Ultraviolet absorbance at 260 and 280 nm in RNA measurement is dependent on measurement solution. AB - RNA measurement is conducted by measuring ultraviolet absorbance at 260 nm and 280 nm. Calculation of the RNA concentration is based on the absorbance at 260 nm. Furthermore, RNA purity is judged as the 260 nm/280 nm ratio and a low ratio indicates contamination by protein. Diethyl-pyrocarbonate (DEPC)-treated water is used to dissolve RNA and 2-amino-2-hydroxymethyl-1,3-propandiol (Tris) is frequently added to the RNA dissolving solution in order to stabilize the RNA. In the present study, RNA was isolated from mouse liver, and then the influence of DEPC-treated water and Tris-buffer on RNA measurement was studied. The 260 nm/280 ratio of RNA determined after diluting it with distilled water was 1.82+/-0.01 (n=5). DEPC-treated water did not affect the absorbance at 260 nm, but elevated that at 280 nm. Thus, the 260 nm/280 nm ratio was as low as 1.52+/-0.01 (n=5). Tris-HCl (1 M, pH 7.0 or 10.0) lowered the absorbance at 260 nm and even more at 280 nm. Thus, the 260 nm/280 nm ratio was elevated to more than 2.17 (n=5). The present results clearly showed the influence of the measurement solution on RNA measurement. PMID- 10812019 TI - Human papillomavirus type 16 E7 impairs the activation of the interferon regulatory factor-1. AB - We had previously observed that HPV-16 E7 disturbs the Guanylate Binding Protein (GBP)-ISRE reporter activation by IFN-gamma thus suggesting an alteration of the IRF-1 function. In this study we examined the mechanism by which E7 affects the IFN-gamma signals driving the activation of gene transcription. Using 14/2 BRK cells containing dexamethasone-inducible HPV-16 E7 gene, we observed a large inhibition of the IRF-1 DNA binding activity upon E7 induction. Concomitantly, there was no significant change in the levels of IRF-1, indicating that this was not due to reduced levels of IRF-1 expression. Likewise, in vitro translated E7 did not affect the IRF-1 DNA binding activity in nuclear extracts derived from IFN-induced cells, thus indicating that the effects of E7 are upstream of IRF-1's binding to its DNA recognition site. Finally, NFkappaB DNA binding activity was also inhibited under conditional expression of E7. These data indicate that HPV 16 E7 inhibits the IRF-1 and NFkappaB function and this could lead to the impairment of the IFN response in HPV-infected cells. Furthermore, the findings suggest that different events of the IFN inducible signal cascade seem to be target for HPV-16 E7. PMID- 10812020 TI - Chrysobalanus icaco L. extract for antiangiogenic potential observation. AB - Angiogenesis is an important process in several physiological situations and it is also implicated in the development of some diseases such as diabetes and cancer. This study investigated the antiangiogenic potential of Chrysobalanus icaco methanol extract in the chicken embrionary tissue. Clinical trials for cancer treatment using drugs based on this mechanism are already in progress. Chorioallantoic membrane model (CAM) of chicken embryos, with C. icaco methanol extract in plastic diskes was used. The results showed an average of 44% angiogenesis inhibition in CAM areas with the plant extract compared to the controls. The data indicate that C. icaco methanol extract reduce the formation of new blood vessels in chicken chorioallantoic membrane. PMID- 10812021 TI - Proton-transfer reactions in bioenergetics. PMID- 10812022 TI - The mechanism of the proton transfer: an outline. AB - A brief summary of the principal notions of the quantum-mechanical theory of the charge transfer reactions has been presented. In the framework of this theory, the mechanism of the proton transfer consists in the classical medium reorganization that equalizes the proton energy levels in the initial and final states, and a consequent proton transfer via a quantum-mechanical underbarrier transition. On the basis of this mechanism, factors influencing the proton transfer probability, and hence kinetic isotope effect, have been discussed; among them are the optimum tunneling distance, the involvement of the excited vibrational states, etc. Semi-classical and quantum-mechanical treatments of the Swain-Schaad relations have been compared. Some applications to enzymatic proton transfer reactions have been described. PMID- 10812023 TI - Calculation of isotope effects from first principles. AB - Various means of calculating the effect of changing the mass of a given atom upon a chemical process are reviewed. Of particular interest is the deuterium isotope effect comparing the normal protium nucleus with its heavier deuterium congener. The replacement of the bridging protium in a neutral hydrogen bond such as the water dimer by a deuterium strengthens the interaction by a small amount via effects upon the vibrational energy. In an ionic H-bond such as the protonated water dimer, on the other hand, the reverse trend is observed in that replacement of the bridging protium by dimer weakens the interaction. In addition to the stability of a given complex, the rate at which a proton transfers from one group to another is likewise affected by deuterium substitution, viz. kinetic isotope effects (KIEs). The KIE is enlarged as the temperature drops, particularly so if the calculation of KIE includes proton tunneling. The KIE is also sensitive to any angular distortions or stretches present in the H-bond of interest. KIEs can be computed either by the standard transition state theory which is derived via only two points on the potential energy surface, or by more complete formalisms which take account of larger swaths of the surface. While more time intensive, the latter can also be applied to provide insights important in interpretation of experimental data. PMID- 10812024 TI - Hydrogen bonds and proton transfer in general-catalytic transition-state stabilization in enzyme catalysis. AB - The question of the nature of the proton bridge involved in general acid-base catalysis in both enzymic and non-enzymic systems is considered in the light of long-known but insufficiently appreciated work of Jencks and his coworkers and of more recent results from neutron-diffraction crystallography and NMR spectroscopic studies, as well as results from isotope-effect investigations. These lines of inquiry lead toward the view that the bridging proton, when between electronegative atoms, is in a stable potential at the transition state, not participating strongly in the reaction-coordinate motion. Furthermore they suggest that bond order is well-conserved at unity for bridging protons, and give rough estimates of the degree to which the proton will respond to structural changes in its bonding partners. Thus if a center involved in general-catalytic bridging becomes more basic, the proton is expected to move toward it while maintaining a unit total bond order. For a unit increase in the pK of a bridging partner, the other partner is expected to acquire about 0.06 units of negative charge. The implications are considered for charge distribution in enzymic transition states as the basicity of catalytic residues changes in the course of molecular evolution or during progress along a catalytic pathway. PMID- 10812025 TI - A pragmatic approach to structure based calculation of coupled proton and electron transfer in proteins. AB - The coupled motion of electrons and protons occurs in many proteins. Using appropriate tools for calculation, the three-dimensional protein structure can show how each protein modulates the observed electron and proton transfer reactions. Some of the assumptions and limitations involved in calculations that rely on continuum electrostatics to calculate the energy of charges in proteins are outlined. Approaches that mix molecular mechanics and continuum electrostatics are described. Three examples of the analysis of reactions in photosynthetic reaction centers are given: comparison of the electrochemistry of hemes in different sites; analysis of the role of the protein in stabilizing the early charge separated state in photosynthesis; and calculation of the proton uptake and protein motion coupled to the electron transfer from the primary (Q(A)) to secondary (Q(B)) quinone. Different mechanisms for stabilizing intra protein charged cofactors are highlighted in each reaction. PMID- 10812026 TI - Marcus rate theory applied to enzymatic proton transfer. AB - The hydration of CO(2) and the dehydration of HCO(3)(-) catalyzed by the carbonic anhydrases is accompanied by the transfer of protons between solution and the zinc-bound water molecule in the active site. This transfer is facilitated by amino acid residues of the enzyme which act as intramolecular proton shuttles; variants of carbonic anhydrase lacking such shuttle residues are enhanced or rescued in catalysis by intermolecular proton transfer from donors such as imidazole in solution. The resulting rate constants for proton transfer when compared with the values of the pK(a) of the donor and acceptor give Bronsted plots of high curvature. These data are described by Marcus theory which shows an intrinsic barrier for proton transfer from 1 to 2 kcal/mol and work terms or thermodynamic contributions to the free energy of reaction from 4 to10 kcal/mol. The interpretation of these Marcus parameters is discussed in terms of the well studied pathway of the catalysis and structure of the enzymes. PMID- 10812027 TI - Common themes and problems of bioenergetics and voltage-gated proton channels. AB - The existence of a proton-selective pathway through a protein is a common feature of voltage-gated proton channels and a number of molecules that play pivotal roles in bioenergetics. Although the functions and structures of these molecules are quite diverse, the proton conducting pathways share a number of fundamental properties. Conceptual parallels include the translocation by hydrogen-bonded chain mechanisms, problems of supply and demand, equivalence of chemical and electrical proton gradients, proton wells, alternating access sites, pK(a) changes induced by protein conformational change, and heavy metal participation in proton transfer processes. An archetypal mechanism involves input and output proton pathways (hydrogen-bonded chains) joined by a regulatory site that switches the accessibility of the bound proton from one 'channel' to the other, by means of a pK(a) change, molecular movement, or both. Although little is known about the structure of voltage-gated proton channels, they appear to share many of these features. Evidently, nature has devised a limited number of mechanisms to accomplish various design strategies, and these fundamental mechanisms are repeated with minor variation in many superficially disparate molecules. PMID- 10812028 TI - Biophysical aspects of intra-protein proton transfer. AB - The passage of proton trough proteins is common to all membranal energy conserving enzymes. While the routes differ among the various proteins, the mechanism of proton propagation is based on the same chemical-physical principles. The proton progresses through a sequence of dissociation association steps where the protein and water molecules function as a solvent that lowers the energy penalty associated with the generation of ions in the protein. The propagation of the proton in the protein is a random walk, between the temporary proton binding sites that make the conducting path, that is biased by the intra protein electrostatic potential. Kinetic measurements of proton transfer reactions, in the sub-ns up to micros time frame, allow to monitor the dynamics of the partial reactions of an overall proton transfer through a protein. PMID- 10812029 TI - Proton transfer reactions across bacteriorhodopsin and along the membrane. AB - Bacteriorhodopsin is probably the best understood proton pump so far and is considered to be a model system for proton translocating membrane proteins. The basis of a molecular description of proton translocation is set by having the luxury of six highly resolved structural models at hand. Details of the mechanism and reaction dynamics were elucidated by a whole variety of biophysical techniques. The current molecular picture of catalysis by BR will be presented with examples from time-resolved spectroscopy. FT-IR spectroscopy monitors single proton transfer events within bacteriorhodopsin and judiciously positioned pH indicators detect proton migration at the membrane surface. Emerging properties are briefly outlined that underlie the efficient proton transfer across and along biological membranes. PMID- 10812030 TI - Proton and electron transfer in bacterial reaction centers. AB - The bacterial reaction center couples light-induced electron transfer to proton pumping across the membrane by reactions of a quinone molecule Q(B) that binds two electrons and two protons at the active site. This article reviews recent experimental work on the mechanism of the proton-coupled electron transfer and the pathways for proton transfer to the Q(B) site. The mechanism of the first electron transfer, k((1))(AB), Q(-)(A)Q(B)-->Q(A)Q(-)(B), was shown to be rate limited by conformational gating. The mechanism of the second electron transfer, k((2))(AB), was shown to involve rapid reversible proton transfer to the semiquinone followed by rate-limiting electron transfer, H(+)+Q(-)(A)Q(-)(B) ifQ( )(A)Q(B)H-->Q(A)(Q(B)H)(-). The pathways for transfer of the first and second protons were elucidated by high-resolution X-ray crystallography as well as kinetic studies showing changes in the rate of proton transfer due to site directed mutations and metal ion binding. PMID- 10812031 TI - Proton pumping by cytochrome oxidase: progress, problems and postulates. AB - The current status of our knowledge about the mechanism of proton pumping by cytochrome oxidase is discussed. Significant progress has resulted from the study of site-directed mutants within the proton-conducting pathways of the bacterial oxidases. There appear to be two channels to facilitate proton translocation within the enzyme and they are important at different parts of the catalytic cycle. The use of hydrogen peroxide as an alternative substrate provides a very useful experimental tool to explore the enzymology of this system, and insights gained from this approach are described. Proton transfer is coupled to and appears to regulate the rate of electron transfer steps during turnover. It is proposed that the initial step in the reaction involves a proton transfer to the active site that is important to convert metal-ligated hydroxide to water, which can more rapidly dissociate from the metals and allow the reaction with dioxygen which, we propose, can bind the one-electron reduced heme-copper center. Coordinated movement of protons and electrons over both short and long distances within the enzyme appear to be important at different parts of the catalytic cycle. During the initial reduction of dioxygen, direct hydrogen transfer to form a tyrosyl radical at the active site seems likely. Subsequent steps can be effectively blocked by mutation of a residue at the surface of the protein, apparently preventing the entry of protons. PMID- 10812032 TI - Where is 'outside' in cytochrome c oxidase and how and when do protons get there? AB - Cytochrome c oxidase moves both electrons and protons in its dual role as a terminal electron acceptor and a contributor to the proton motive force which drives the formation of ATP. Although the sequence of electron transfer events is well-defined, the correlated mechanism and routes by which protons are translocated across the membrane are not. A recent model [Michel, Proc. Natl. Acad. Sci. USA 95 (1998) 12819] offers a detailed molecular description of when and how protons are translocated through the protein to the outside, which contrasts with previous models in several respects. This article reviews the behavior of site-directed mutants of Rhodobacter sphaeroides cytochrome c oxidase in the context of these different models. Studies of the internally located lysine 362 on the K channel and aspartate 132 on the D channel, indicate that D132, but not K362, is connected to the exterior region. Analysis of the externally located arginine pair, 481 and 482, and the Mg/Mn ligands, histidine 411 and aspartate 412, which are part of the hydrogen-bonded network that includes the heme propionates, indicates that alterations in this region do not strongly compromise proton pumping, but do influence the pH dependence of overall activity and the control of activity by the pH gradient. The results are suggestive of a region of 'sequestered' protons: beyond a major energetic gate, but selectively responsive to the external environment. PMID- 10812033 TI - Mechanism of proton translocation by cytochrome c oxidase: a new four-stroke histidine cycle. PMID- 10812034 TI - Proton and hydrogen currents in photosynthetic water oxidation. AB - The photosynthetic processes that lead to water oxidation involve an evolution in time from photon dynamics to photochemically-driven electron transfer to coupled electron/proton chemistry. The redox-active tyrosine, Y(Z), is the component at which the proton currents necessary for water oxidation are switched on. The thermodynamic and kinetic implications of this function for Y(Z) are discussed. These considerations also provide insight into the related roles of Y(Z) in preserving the high photochemical quantum efficiency in Photosystem II (PSII) and of conserving the highly oxidizing conditions generated by the photochemistry in the PSII reaction center. The oxidation of Y(Z) by P(680)(+) can be described well by a treatment that invokes proton coupling within the context of non adiabatic electron transfer. The reduction of Y(.)(Z), however, appears to proceed by an adiabatic process that may have hydrogen-atom transfer character. PMID- 10812035 TI - Exaggerated amygdala response to masked facial stimuli in posttraumatic stress disorder: a functional MRI study. AB - BACKGROUND: Converging lines of evidence have implicated the amygdala in the pathophysiology of posttraumatic stress disorder (PTSD). We previously developed a method for measuring automatic amygdala responses to general threat-related stimuli; in conjunction with functional magnetic resonance imaging, we used a passive viewing task involving masked presentations of human facial stimuli. METHODS: We applied this method to study veterans with PTSD and a comparison cohort of combat-exposed veterans without PTSD. RESULTS: The findings indicate that patients with PTSD exhibit exaggerated amygdala responses to masked-fearful versus masked-happy faces. CONCLUSIONS: Although some previous neuroimaging studies of PTSD have demonstrated amygdala recruitment in response to reminders of traumatic events, this represents the first evidence for exaggerated amygdala responses to general negative stimuli in PTSD. Furthermore, by using a probe that emphasizes automaticity, we provide initial evidence of amygdala hyperresponsivity dissociated from the "top-down" influences of medial frontal cortex. PMID- 10812036 TI - Research on eating disorders: current status and future prospects. AB - This report summarizes the main themes addressed at a workshop on research on eating disorders (EDs), which was hosted by the National Institute of Mental Health in December 1998. Both basic behavioral neuroscientists and clinical investigators met in an effort to integrate areas of research and foster collaborations. Considerable advances have been made in understanding the neuroendocrinological mechanisms that regulate appetite and food intake. These achievements are in sharp contrast with the limited progress in elucidating the pathogenesis of EDs and developing effective treatment and preventive interventions. Anorexia nervosa remains a highly morbid condition with the highest mortality of any other psychiatric disorder. Besides acute refeeding techniques, no specific interventions have been proven effective in changing the long-term course of anorexia nervosa. Efficacious treatments exist for bulimia nervosa, but their underutilization calls for research on translating experimental findings into clinical practice. Closer interface between neuroscientists and clinical researchers is required for advancing our understanding of ED pathogenesis and developing effective treatments. Recent studies are suggestive of a substantial genetic contribution to EDs that deserves further investigation. Finally, there is an urgent need to examine risk and protective factors for EDs, on which safe and effective prevention can be built. PMID- 10812037 TI - Psychiatric disorders and mutations at the Wolfram syndrome locus. AB - Identifying genetic loci at which mutations predispose individuals to common psychiatric illnesses will have major impact on the diagnosis and treatment of mental illness. The available evidence indicates that mutations at the Wolfram syndrome locus contribute substantially to the prevalence of psychiatric illness in the general population. Patients with mutations at this locus on both parental chromosomes, called Wolfram syndrome homozygotes, have a distinctive and rare autosomal recessive syndrome characterized by juvenile onset diabetes mellitus and bilateral progressive optic atrophy. Diverse and serious psychiatric manifestations frequently have been observed in Wolfram syndrome patients; however, the population burden of mental illness attributable to mutations at this locus is almost entirely from individuals who carry a single mutation, called Wolfram syndrome heterozygotes, who have no distinguishing physical characteristics but constitute approximately 1% of the population. Molecular genotyping of blood relatives of Wolfram syndrome patients has shown that Wolfram syndrome heterozygotes are 26-fold more likely than noncarriers to have a psychiatric hospitalization. Severe depression was the predominant finding in the test group studied. The prediction that approximately 25% of all patients hospitalized for depression are Wolfram syndrome heterozygotes now can be tested by mutation screening of hospitalized patients from the general population. Many other behavioral and cognitive difficulties also have been observed in Wolfram syndrome families. For each specific psychiatric abnormality, a "test group" of blood relatives within Wolfram syndrome families with that abnormality can be formed. By comparing the number of Wolfram syndrome heterozygotes found in each test group by molecular genotyping with the number expected under the null hypothesis, the index-test method can determine which clinical phenotypes result from mutations at the Wolfram syndrome locus. This method can be utilized to identify other loci at which mutations predispose individuals to psychiatric illnesses. PMID- 10812038 TI - A search for susceptibility loci for anorexia nervosa: methods and sample description. AB - BACKGROUND: Eating disorders have not traditionally been viewed as heritable illnesses; however, recent family and twin studies lend credence to the potential role of genetic transmission. The Price Foundation funded an international, multisite study to identify genetic factors contributing to the pathogenesis of anorexia nervosa (AN) by recruiting affective relative pairs. This article is an overview of study methods and the clinical characteristics of the sample. METHODS: All probands met modified DSM-IV criteria for AN; all affected first, second, and third degree relatives met DSM-IV criteria for AN, bulimia nervosa (BN), or eating disorder not otherwise specified (NOS). Probands and affected relatives were assessed diagnostically with the Structured Interview for Anorexia and Bulimia. DNA was collected from probands, affected relatives and a subset of their biological parents. RESULTS: Assessments were obtained from 196 probands and 237 affected relatives, over 98% of whom are of Caucasian ancestry. Overall, there were 229 relative pairs who were informative for linkage analysis. Of the proband-relative pairs, 63% were AN-AN, 20% were AN-BN, and 16% were AN-NOS. For family-based association analyses, DNA has been collected from both biological parents of 159 eating-disordered subjects. Few significant differences in demographic characteristics were found between proband and relative groups. CONCLUSIONS: The present study represents the first large-scale molecular genetic investigation of AN. Our successful recruitment of over 500 subjects, consisting of affected probands, affected relatives, and their biological parents, will provide the basis to investigate genetic transmission of eating disorders via a genome scan and assessment of candidate genes. PMID- 10812039 TI - Lactate-induced rage and panic in a select group of subjects who perpetrate acts of domestic violence. AB - BACKGROUND: Perpetrators of domestic violence frequently report symptoms of autonomic arousal and a sense of fear and/or loss of control at the time of the violence. Since many of these symptoms are also associated with panic attacks, we hypothesized that perpetrators of domestic violence and patients with panic attacks may share similar exaggerated fear-related behaviors. To test this hypothesis, we employed the panicogenic agent sodium lactate to examine the response of perpetrators to anxiety fear induced by a chemical agent. METHODS: Using a double-blind, placebo-controlled design, we infused 0.5 mol/L sodium lactate or placebo over 20 min on separate days to a select group of subjects who perpetrate acts of domestic violence and two nonviolent comparison groups. We compared their behavioral, neuroendocrine, and physiologic responses. RESULTS: Lactate administration elicited intense emotional responses in the perpetrators of domestic violence. Perpetrators evidenced more lactate-induced rage and panic and showed greater changes in speech, breathing, and motor activity than did nonviolent control subjects. There were no significant differences between the groups for any neuroendocrine or physiologic measure. CONCLUSIONS: These results are consistent with our hypothesis that some perpetrators of domestic violence have exaggerated fear-related behavioral responses. PMID- 10812040 TI - A randomized double-blind fluvoxamine/placebo crossover trial in pathologic gambling. AB - BACKGROUND: The study assessed the efficacy and tolerability of the selective serotonin reuptake inhibitor (SSRI) fluvoxamine in the treatment of pathologic gambling (PG). METHODS: A 16-week randomized double-blind crossover design insured that each subject received 8 weeks of fluvoxamine and 8 weeks of a placebo. Fifteen patients entered and 10 subjects, all male, completed the study. RESULTS: Fluvoxamine resulted in a significantly greater percent improvement in overall gambling severity on the PG Clinical Global Impression (PG-CGI) scale. There was a significant drug effect on gambling urge and behavior as measured by the PG modification of the Yale-Brown Obsessive Compulsive Scale and PG-CGI scale improvement scores; however, there was a significant interaction of drug effect with the order of administration of drug and placebo. Post hoc analysis, treating each phase as a separate trial, demonstrated a significant difference between fluvoxamine and the placebo in the second phase of the trial but not in the first. Fluvoxamine side effects were of only mild intensity and consistent with SSRI treatment and were not associated with early withdrawal from the study. CONCLUSIONS: These findings suggest that fluvoxamine is well tolerated and may be effective in the treatment of PG in an acute trial, and that an early placebo effect in PG treatment appears to diminish over time. To confirm this finding and to determine whether improvement persists over an extended period of time, a longer duration parallel-design trial with long-term maintenance follow-up should be conducted in a larger and more diverse PG population. PMID- 10812041 TI - Reboxetine: a pharmacologically potent, selective, and specific norepinephrine reuptake inhibitor. AB - BACKGROUND: Reboxetine is a potent antidepressant, with efficacy comparable to that of imipramine, desipramine, and fluoxetine, and has improved side-effect profile. The basis of its efficacy and improved tolerability is sought through studies of reboxetine in a number of pharmacological models of depression. METHODS: Pharmacological selectivity for uptake systems was defined by uptake and binding assays for the three monoamine uptake sites. Specificity was determined in 39 different receptor and 6 enzyme assays. In vivo selectivity was defined by measurement of neuronal firing rates in the locus coeruleus, dorsal raphe, and substantia nigra. Reserpine-induced blepharospasm and hypothermia, clonidine induced hypothermia, defined reboxetine's in vivo pharmacology. Reboxetine's antidepressant potential was evaluated behaviorally by the tail-suspension test, forced swimming, and the DRL72 operant responding test. RESULTS: Reboxetine is a potent, selective, and specific norepinephrine reuptake inhibitor (selective NRI) as determined by both in vitro and in vivo measurements. Unlike desipramine or imipramine, reboxetine has weak affinity (Ki > 1,000 nmol/L)for muscarinic, histaminergic H1, adrenergic alpha1, and dopaminergic D2 receptors. In vivo action of reboxetine is entirely consistent with the pharmacological action of an antidepressant with preferential action at the norepinephrine reuptake site. Reboxetine showed an antidepressant profile in all tests of antidepressant activity used. Significant decreases in immobility were observed in the tail suspension test and behavioral despair test. Increased efficiency in responding was observed in the DRL72 test. CONCLUSIONS: Reboxetine is a potent, selective, and specific noradrenergic reuptake inhibitor. It has a superior pharmacological selectivity to existing tricyclic antidepressants and selective serotonin reuptake inhibitors when tested in a large number of in vitro and in vivo systems. Given the pharmacological profile, reboxetine is expected to be a selective and potent tool for psychopharmacological research. The use of reboxetine in the clinic will also help clarify the role norepinephrine plays in depression. PMID- 10812042 TI - Reactivity to a 35% CO2 challenge in healthy first-degree relatives of patients with panic disorder. AB - BACKGROUND: The effects of a 35% CO2 challenge were examined in healthy first degree relatives of panic disorder patients and in healthy control subjects matched for age and gender. METHODS: One single inhalation of a 35% CO2/65% O2 challenge was administered to 50 first-degree relatives of panic disorder patients and 50 control subjects. RESULTS: The first-degree relatives were more reactive to the 35% CO2 challenge than the control subjects. CONCLUSIONS: These findings indicate that being a member of a family with a panic disorder patient is, in itself an important factor in CO2 hypersensitivity among subjects who have never experienced a panic attack. Both panic disorder patients and their first degree relatives have a tendency to be more reactive to the CO2 challenge. PMID- 10812043 TI - Fluoxetine prevents PCP- and MK801-induced HSP70 expression in injured limbic cortical neurons of rats. AB - BACKGROUND: N-Methyl-D-aspartate (NMDA) receptor antagonists, including phencyclidine (PCP) and dizocilpine (MK801), cause schizophrenialike psychosis in humans, and produce vacuolated neurons in the cingulate and retrosplenial cortices of the rat brain. Since psychotically depressed patients and schizophrenic depressed patients may require treatment with selective serotonin reuptake inhibitors (SSRIs), it is of interest to examine the relationship between SSRIs and NMDA antagonist neurotoxicity. METHODS: The neurotoxicity of PCP and MK801 was assessed using heat shock protein (HSP70) immunocytochemistry and HSP70 Western blots because HSP70 is expressed in the injured, vacuolated neurons. Female rats were given fluoxetine (0, 5, 10, and 20 mg/kg IP) followed 1 hour later by MK801 (1 mg/kg IP) or PCP (50 mg/kg IP). RESULTS: Pretreatment with fluoxetine (20 mg/kg IP) 1 hour before MK801 prevented the induction of HSP70 by MK801 in the cingulate and retrosplenial cortices. Pretreatment with fluoxetine (10 or 20 mg/kg IP) 1 hour before PCP also prevented the HSP70 induction by PCP. CONCLUSIONS: Fluoxetine prevents the neurotoxicity of NMDA receptor antagonists in rat brain. This suggests the possibility that SSRIs could modulate psychosis, and may provide a model for examining the link between the hallucinogenic properties of PCP and lysergic acid diethylamide. PMID- 10812044 TI - Paroxetine binding to the rat norepinephrine transporter in vivo. AB - BACKGROUND: The norepinephrine transporter (NET)/uptake site is an antidepressant sensitive transporter located on plasma membranes of noradrenergic neurons and other specialized cells that remove norepinephrine (NE) from the synapse to terminate the actions of NE. The antidepressant paroxetine is believed to produce its therapeutic effects primarily by acting as a highly selective antagonist of the serotonin transporter (SERT). However, in vitro data indicates that paroxetine inhibits the NET. The present study was designed to determine whether paroxetine inhibits in NET in vivo. METHODS: Rats were administered paroxetine (6.5, 10.0, or 15.0 mg/kg/day) via osmotic minipumps for 1 week. Following attainment of steady state serum concentrations, cortical NET function was assessed by both [3H]-nisoxetine binding and [3H]-norepinephrine uptake assays conducted ex vivo. RESULTS: In unwashed brain homogenates, serum paroxetine concentrations greater than 100 ng/mL were positively correlated with the observed Kd for [3H]-nisoxetine. At [3H]-nisoxetine concentrations associated with 50% transporter occupancy in vehicle treated rats, [3H]-nisoxetine binding was decreased 21% and 34% in rats exhibiting serum paroxetine concentrations > 100 ng/mL and > 500 ng/mL, respectively. CONCLUSIONS: Although paroxetine is a very potent inhibitor of the SERT, paroxetine also inhibits the NET at serum concentrations > 100 ng/mL. This novel finding may underlie the broad therapeutic utility of paroxetine in mood and anxiety disorders. PMID- 10812046 TI - Complex actions of protein kinase A inhibitors on mitogenesis of bovine coronary artery smooth muscle cells. AB - This study investigates the possible modulation of platelet-derived growth factor (PDGF, 20 ng/ml)-induced DNA synthesis in bovine coronary artery smooth muscle cells by the protein kinase A inhibitors Rp-adenosine-3',5'-cyclic phosphorothioate (Rp-cAMPS, 0. 03-10 microM) and ?N-[2-((p bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, HCl? (H-89, 0.01-1 microM). Rp-cAMPS concentration dependently enhanced PDGF-induced DNA synthesis. In contrast, no potentiation of PDGF-induced DNA synthesis was seen with H-89. However, H-89 but not Rp-cAMPS, inhibited p42/p44 mitogen-activated protein kinase phosphorylation. Thus, Rp-cAMPS, but not H-89, unmasks inhibitory actions of protein kinase A on PDGF-induced mitogenesis of vascular smooth muscle cells. Low specificity may limit the use of H-89 as protein kinase A inhibitor. PMID- 10812047 TI - Hydrogen peroxide generation by monoamine oxidases in rat white adipocytes: role on cAMP production. AB - In rat, white adipocytes monoamine oxidases (EC 1.4.3.4.) generate hydrogen peroxide (H(2)O(2)). Recent studies suggested that, in addition to its toxic features, H(2)O(2) may behave as a cell second messenger. In the present study, using fluorimetric and chemiluminescence (CL) assays, we showed that tyramine degradation by monoamine oxidases in intact adipocytes resulted in the concentration-dependent generation of H(2)O(2). In addition, we found that, in the presence of low tyramine concentrations, forskolin-dependent cAMP production was significantly increased as compared to that of the control and this increase was prevented by the monoamine oxidase inhibitor pargyline or by the H(2)O(2) trapping system homovanillic acid-peroxidase. Finally, we demonstrated that tyramine degradation by monoamine oxidases increased the ability of isoproterenol to induce cell lipolysis. Taken together, these data suggest that H(2)O(2) produced during substrate degradation by monoamine oxidases may participate in the regulation of adipocyte metabolism. PMID- 10812048 TI - Inhibition of phospholipase A(2) activity by S-nitroso-cysteine in a cyclic GMP independent manner in PC12 cells. AB - Arachidonic acid and nitric oxide (NO) act as retrograde and intercellular messengers in the nervous system. Regulation of cyclooxygenase is well established, but regulation of phospholipase A(2), the enzyme responsible for the liberation of arachidonic acid, by NO has not been thoroughly investigated. Using the PC12 cell line as a neuronal model, we studied the effects of exogenous NO compounds on arachidonic acid release. Incubation with Ca(2+) ionophores or mastoparan (wasp venom peptide) stimulated [3H]arachidonic acid release from prelabeled PC12 cells. [3H]Arachidonic acid release was inhibited by cytosolic phospholipase A(2) inhibitors, but not by dithiothreitol. A cytosolic phospholipase A(2) protein band with a molecular mass of approximately 100 kDa was detected by immunoblotting. S-Nitroso-cysteine inhibited basal and stimulated [3H]arachidonic acid release in concentration-dependent manners. Other NO compounds such as sodium nitroprusside and S-nitroso-N-acetylpenicillamine did not affect [3H]arachidonic acid release. N-Ethylmaleimide also inhibited [3H]arachidonic acid release. The inhibitory effects of S-nitroso-cysteine and N ethylmaleimide were irreversible, because [3H]arachidonic acid release from PC12 cells preincubated with S-nitroso-cysteine or N-ethylmaleimide was much lower than that from nontreated cells. These findings suggest (a) cytosolic phospholipase A(2) is activated by Ca(2+) or mastoparan, and inhibited by S nitroso-cysteine in a cyclic GMP-independent manner, (b) N-ethylmaleimide also inhibits cytosolic phospholipase A(2) and arachidonic acid release in PC12 cells. S-Nitroso-cysteine can regulate the production of other retrograde messenger arachidonic acid. PMID- 10812049 TI - Effect of bioflavonoids on vincristine transport across blood-brain barrier. AB - Several grapefruit juice bioflavonoids, including quercetin, are reported to stimulate P-glycoprotein-mediated drug efflux from cultured tumor cells. To see whether these bioflavonoids alter the permeation of vincristine across the blood brain barrier, we conducted experiments with cultured mouse brain capillary endothelial cells (MBEC4 cells) in vitro and ddY mice in vivo. The steady-state uptake of [3H]vincristine by MBEC4 cells was decreased by 10 microM quercetin, but increased by 50 microM quercetin. Similarly, the in vivo brain-to-plasma concentration ratio of [3H]vincristine in ddY mice was decreased by coadministration of 0.1 mg/kg quercetin, but increased by 1.0 mg/kg quercetin. Kaempferol had a similar biphasic effect on the in vitro uptake of [3H]vincristine. Other aglycones tested (chrysin, flavon, hesperetin, naringenin) increased [3H]vincristine uptake in the 10-50 microM range, and glycosides (hesperidin, naringin, rutin) were without effect. We then addressed the mechanism of the concentration-dependent biphasic action of quercetin. Verapamil, a P-glycoprotein inhibitor, inhibited the efflux of [3H]vincristine from MBEC4 cells, while 10 microM quercetin significantly stimulated it. The uptake of [3H]vincristine by MBEC4 cells was increased by inhibitors of protein kinase C, but decreased by phorbol 12-myristate-13-acetate (PMA), as well as by 10 microM quercetin. The phosphorylation level of P-glycoprotein was increased in the presence of 5 microM quercetin or 100 nM PMA, but decreased by the protein kinase C inhibitor H7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine, 30 microM). We conclude that low concentrations of quercetin indirectly activate the transport of [3H]vincristine by enhancing the phosphorylation (and hence activity) of P glycoprotein, whereas high concentrations of quercetin inhibit P-glycoprotein. Our results indicate that patients taking drugs which are P-glycoprotein substrates may need to restrict their intake of bioflavonoid-containing foods and beverages, such as grapefruit juice. PMID- 10812050 TI - Interaction between intrathecal morphine and glutamate receptor antagonists in formalin test. AB - The analgesic interaction between intrathecally administered morphine and the NMDA receptor antagonist, ((+/-)-2-amino-5-phosphonopentanoic acid; AP-5), the NMDA receptor glycine site antagonist, (5-nitro-6,7-dichloro-2,3-quinoxaline dion; ACEA 1021), or the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid) receptor antagonist (ACEA 2752) in the formalin test was investigated with a rat model of chronic lumbar intrathecal catheterization. After obtaining dose-response curves for each agent, combinations of morphine and AP-5, ACEA 1021 or ACEA 2752 were tested for their effect on the number of flinches in the formalin test and for associated side-effects, such as motor disturbances, flaccidity, and agitation/allodynia. Using isobolographic analyses, a potent analgesic synergy was observed with decreased side-effects between morphine and ACEA 2752 or AP-5. ACEA 1021 increased the analgesic effect of low dose morphine. Spinal mu-opioid receptor activation and NMDA or AMPA receptor antagonism showed a synergistic antinociception against tonic pain. These results suggest an important direction in the management of inflammatory pain. PMID- 10812051 TI - Effects of endomorphins-1 and -2, endogenous mu-opioid receptor agonists, on spontaneous alternation performance in mice. AB - The effects of intracerebroventricular (i.c.v.) administration of endomorphins-1 and -2, endogenous mu-opioid receptor agonists, on the spontaneous alternation performance associated with spatial working memory were investigated in mice. Endomorphin-1 (10 and 17.5 microg) and endomorphin-2 (10 microg) produced a significant decrease in percent alternation without affecting total arm entries. beta-Funaltrexamine (5 microg) almost completely reversed the endomorphin-1 (10 microg)- and endomorphin-2 (10 microg)-induced decrease in percent alternation, although neither naltrindole (4 ng) nor nor-binaltorphimine (4 microg) produced any significant effects on alternation performance. These results suggest that endomorphins impair spatial working memory through the mediation of mu-opioid receptors. PMID- 10812052 TI - Beneficial effects of MET-88, a gamma-butyrobetaine hydroxylase inhibitor in rats with heart failure following myocardial infarction. AB - Myocardial ischemia can cause myocardial infarction and as a consequence, heart failure. 3-(2,2,2-trimethylhydrazinium) propionate (MET-88) inhibits gamma butyrobetaine hydroxylase and has cardioprotective effects on the ischemic heart. We now examined the effects of MET-88 in rats with congestive heart failure following myocardial infarction. Congestive heart failure was produced by left coronary artery ligation in rats. MET-88 at 100 mg/kg/day was orally administered from the 2nd day after surgery. We performed a survival study for 181 days, and measured ventricular remodeling, cardiac function, and myocardial high-energy phosphate levels after treatment for 20 days. MET-88 prolonged survival with a median 50% survival of 103 days compared to 79 days for the heart-failure control rats. The expansion of the left ventricular cavity (ventricular remodeling) in heart-failure rats was prevented by treatment with MET-88, and the effect of MET 88 was similar to that of captopril at 20 mg/kg. MET-88 attenuated the rise in right atrial pressure in heart-failure rats and augmented cardiac functional adaptability against an increased load. Also, MET-88 improved the myocardial energy state in heart-failure rats. The present results indicate that MET-88 improves the pathosis in rats with heart failure induced by myocardial infarction. PMID- 10812053 TI - Endothelial reactivity to the immediate hypersensitivity reaction of guinea pig pulmonary artery. AB - Ovalbumin at low doses (0.1 microg/ml) caused pronounced relaxations in the precontracted pulmonary arteries of sensitized guinea pigs but, at high doses, (1 100 microg/ml) the relaxations were blunted by the contractions. The relaxations in response to ovalbumin challenge were related to histamine, which is released during the immediate hypersensitivity reaction, because they were almost blocked by mepyramine (10(-5) M) plus cimetidine (10(-4) M) pretreatment and never observed in unsensitized animal arteries. Additionally, the inhibition of relaxations by endothelium removal or N(G)-nitro-L-arginine (L-NOARG, 10(-4) M) treatment implies that the phenomenon requires endothelial nitric oxide synthesis. However, the contractions appear to depend on leukotriene production since they were markedly blocked in the presence of 2(S)-hydroxy-3(R)-[(2 carboxyethyl)thio]-3-[2-(8-phenyloctyl)pheny l]- propanoic acid (SKF 104353, 10( 5) M), a leukotriene receptor antagonist. These results indicate that ovalbumin induced nitric oxide and histamine H(2) receptor dependent relaxations in pulmonary artery may have an important role in the recovery of the increased pulmonary vascular resistance during the hypersensitivity reaction. PMID- 10812054 TI - Picroliv preconditioning protects the rat liver against ischemia-reperfusion injury. AB - Cell death following ischemia-reperfusion injury is a major concern in clinical issues such as organ transplantation and trauma. The need to identify agents with a potential for preventing such damage has assumed great importance. We have evaluated the efficacy of picroliv, a potent antioxidant derived from the plant Picrorhiza kurrooa, in protecting against hepatic ischemia-reperfusion injury in vivo. Picroliv was fed to male Sprague Dawley rats in a dose of 12 mg/kg once daily by oral gavage for 7 days prior to hepatic ischemia. Ischemia was induced by occluding the hepatic pedicel with a microaneurysm clip for 30 min and reperfusion was allowed thereafter for varying period (15-120 min) by releasing the microaneurysm clip. Picroliv pretreatment resulted in better hepatocyte glycogen preservation and reduced apoptosis. Reduction in apoptosis was associated with decreased mRNA expression of caspase-3 and Fas. Oxidant induced cellular damage as measured by tissue malondialdehyde (MDA) levels was significantly less following picroliv pretreatment. Both a reduction in neutrophil infiltration and an increased level of intracellular antioxidant enzyme superoxide dismutase possibly contributed to the reduction in tissue lipid peroxidation. Tissue inflammatory cytokines level of interleukin-1alpha (IL 1alpha) and interleukin-1beta (IL-1beta) was also lower in picroliv group. Furthermore, picroliv pretreatment resulted in enhanced proliferating cell nuclear antigen (PCNA) immunoreactivity. These studies strongly suggest picroliv to be a promising agent for ameliorating injury following ischemia-reperfusion. PMID- 10812055 TI - Effects of TAK-637, a tachykinin receptor antagonist, on the micturition reflex in guinea pigs. AB - The effects of a new tachykinin NK(1) receptor antagonist, (aR, 9R)-7-[3,5 bis(trifluoromethyl)benzyl]-8,9,10, 11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H [1,4]diazocino[2,1-g] [1, 7]naphthyridine-6,13-dione (TAK-637), on the micturition reflex were compared with those of drugs used for abnormally frequent micturition or incontinence. TAK-637 showed a characteristic effect on the distension-induced rhythmic bladder contractions in guinea pigs. The systemic administration of TAK-637 decreased the number but not the amplitude of the distension-induced rhythmic bladder contractions. A similar effect was observed in animals in which the spinal cord had been severed. TAK-637 also inhibited the micturition reflex induced by topical application of capsaicin onto the surface of bladder dome. From these results, it is concluded that TAK-637 inhibits sensory transmissions from the bladder evoked by both physiological and nociceptive stimuli by blocking tachykinin NK(1) receptors, possibly at the level of the spinal cord. On the other hand, the other drugs such as oxybutynin, tolterodine, propiverine, and inaperisone showed no effects on the frequency of the distension-induced rhythmic bladder contractions but decreased the contraction amplitude. Therefore, TAK-637 may represent a new class of drugs, which would be effective for abnormally frequent micturition without causing voiding difficulties due to decreased voiding pressure. PMID- 10812056 TI - Bradykinin potentiates prostaglandin E(2) release in the human gingival fibroblasts pretreated with interleukin-1beta via Ca(2+) mobilization. AB - Interleukin-1beta, a proinflammatory cytokine, causes a slow increase in prostaglandin E(2) release. On the other hand, bradykinin, a chemical mediator for inflammation, induces a rapid prostaglandin E(2) release. Simultaneous stimulation with interleukin-1beta (200 pg/ml) and bradykinin (1 microM) evoked a moderately synergistic increase in prostaglandin E(2) release in human gingival fibroblasts. However, in the human gingival fibroblasts pretreated with interleukin-1beta, bradykinin drastically enhanced prostaglandin E(2) release. NS 398, a specific inhibitor of cyclooxygenase-2, inhibited not only interleukin 1beta-induced prostaglandin E(2) release but also bradykinin-induced prostaglandin E(2) release in the human gingival fibroblasts pretreated with interleukin-1beta. Transcriptional and translational inhibitors such as actinomycin D, cycloheximide, and dexamethasone also suppressed the interleukin 1beta-induced prostaglandin E(2) release and the bradykinin-induced prostaglandin E(2) release in interleukin-1beta-pretreated human gingival fibroblasts. In the fibroblasts pretreated with interleukin-1beta, Ca(2+)-mobilizing reagents such as ionomycin and thapsigargin mimicked the potentiating effect of bradykinin on prostaglandin E(2) release. These results suggest that interleukin-1beta- and bradykinin-induced prostaglandin E(2) release is dependent on cyclooxygenase-2 and the potentiated effect of bradykinin in the human gingival fibroblasts primed with interleukin-1beta is caused by Ca(2+) mobilization. PMID- 10812057 TI - Preferential inhibition of cyclooxygenase-2 by meloxicam in human rheumatoid synoviocytes. AB - The aim of this study was to evaluate the anti-inflammatory effect of 4-hydroxy-2 methyl-N-[5-methyl-2-thiazolyl]-2H-1, 2-benzothiazine-3-carboxamide-1,1-dioxide (meloxicam) using cultured rheumatoid synovial fibroblast-like cells (synoviocytes). Synoviocytes were treated with meloxicam in the presence or absence of interleukin-1beta. Meloxicam had no effect on both cyclooxygenase-1 and -2 expression as determined by Western blot analysis, immunohistochemical staining, and reverse transcription polymerase chain reaction (RT-PCR). Even the lower doses of meloxicam inhibited cyclooxygenase-2 activity, but only the higher doses of meloxicam inhibited cyclooxygenase-1 activity as determined by prostaglandin E(2) synthesis assay. So meloxicam had a preferential inhibitory effect of cyclooxygenase-2 relative to cyclooxygenase-1 on cultured rheumatoid synoviocytes without affecting cyclooxygenase expression. On the other hand, indomethacin had no selectivity and dexamethasone inhibited the expression of cyclooxygenase-2. Our data indicate that clinical efficacy and safety of meloxicam for rheumatoid arthritis may result from its preferential inhibition of cyclooxygenase-2 activity relative to cyclooxygenase-1 on rheumatoid synoviocytes. PMID- 10812058 TI - alpha-Helical solenoid model for the human involucrin. AB - Involucrin is a key component of the cross-linked envelope of terminally differentiated keratinocytes. The human molecule largely consists of 10 residue repeats and forms a thin 460 A long rod. Summarized experimental data and a detailed stereochemical analysis made with computer modeling resulted in a structural model for the involucrin molecule. The suggested structure is a left handed alpha-helical solenoid built of a tandem array of helix-turn-helix folds. The structure enables us to explain the whole set of experimental data and residue conservations within the repeats. It is ideally suited to serve as a scaffold for cell envelope assembly and proposes a possible mode of the intermolecular interactions of involucrin during cell cornification. PMID- 10812059 TI - Endogenous expression of the beta1A sodium channel subunit in HEK-293 cells. AB - The expression of the sole alpha-subunit of muscle or brain sodium channels in frog oocytes mediates currents with a bimodal inactivation with an abnormal slow mode that is strongly depressed only by co-expression of the beta1-subunit. In contrast, in the expression of the alpha-subunit in the human embryonic kidney cell line, HEK-293, the slow mode is almost absent, suggesting an endogenous expression of the beta1-subunit. We have tested this hypothesis by reverse transcriptase-polymerase chain reaction. We found an abundant expression of mRNA encoding the beta1A splicing of the putative regulatory sodium channel subunit but no mRNA encoding the beta1-subunit in HEK cells. This finding is consistent with the idea that the endogenous beta1A-subunit is sufficient for suppressing the slowly inactivating mode of sodium currents by co-assembly with alpha subunits, and calls attention to the reliability of effects attributed in HEK cells to alpha-beta1 co-expression. PMID- 10812060 TI - The SAAS granin exhibits structural and functional homology to 7B2 and contains a highly potent hexapeptide inhibitor of PC1. AB - Prohormone convertases (PCs) 1 and 2 are thought to mediate the proteolytic cleavage of many peptide precursors. Endogenous inhibitors of both PC1 and PC2 have now been identified; the 7B2 protein is a nanomolar inhibitor of PC2, while the novel protein proSAAS was recently reported to be a micromolar inhibitor of PC1 [Fricker et al. (2000) J. Neurosci. 20, 639-648]. We here report evidence that 7B2 and proSAAS exhibit several elements of structural and functional homology. Firstly, 26 kDa human, mouse and rat proSAAS, like all vertebrate 7B2s, contain a proline-rich sequence within the first half of the molecule and also contain a C-terminal 40 residue peptide (SAAS CT peptide) separated from the remainder of the protein by a furin consensus sequence. The SAAS CT peptide contains the precise sequence of a hexapeptide previously identified by combinatorial peptide library screening as a potent inhibitor of PC1, and the vast majority of the inhibitory potency of proSAAS can be attributed to this hexapeptide. Further, like the 7B2 CT peptide, SAAS CT-derived peptides represent tight-binding competitive convertase inhibitors with nanomolar potencies. Lastly, recombinant PC1 is able to cleave the proSAAS CT peptide to a product with a mass consistent with cleavage following the inhibitory hexapeptide. Taken together, our results indicate that proSAAS and 7B2 may comprise two members of a functionally homologous family of convertase inhibitor proteins. PMID- 10812061 TI - Highly divergent sequences of the pollen self-incompatibility (S) gene in class-I S haplotypes of Brassica campestris (syn. rapa) L. AB - Self-incompatibility (SI) enables flowering plants to discriminate between self- and non-self-pollen. In Brassica, SI is controlled by the highly polymorphic S locus. The recently identified male determinant, termed SP11 or SCR, is thought to be the ligand of S receptor kinase, the female determinant. To examine functional and evolutionary properties of SP11, we cloned 14 alleles from class-I S haplotypes of Brassica campestris and carried out sequence analyses. The sequences of mature SP11 proteins are highly divergent, except for the presence of conserved cysteines. The phylogenetic trees suggest possible co-evolution of the genes encoding the male and female determinants. PMID- 10812062 TI - Flavonoids can replace alpha-tocopherol as an antioxidant. AB - Endogenous antioxidants such as the lipid-soluble vitamin E protect the cell membranes from oxidative damage. Glutathione seems to be able to regenerate alpha tocopherol via a so-called free radical reductase. The transient protection by reduced glutathione (GSH) against lipid peroxidation in control liver microsomes is not observed in microsomes deficient in alpha-tocopherol. Introduction of antioxidant flavonoids, such as 7-monohydroxyethylrutoside, fisetin or naringenin, into the deficient microsomes restored the GSH-dependent protection, suggesting that flavonoids can take over the role of alpha-tocopherol as a chain breaking antioxidant in liver microsomal membranes. PMID- 10812063 TI - Suppressive interactions between mutations located in the two nucleotide binding domains of CFTR. AB - The S1235R locus in CFTR was studied in combination with alleles found at the M470V and G628R loci. While R628 caused a maturational defect, R1235 did not. The impact of R1235 was found to be influenced by the alleles present at the G628R and M470V loci. At the single channel level, R1235-V (R1235 on a V470 background) was characterized by an open probability significantly higher than V470-wildtype CFTR. M470, which on its own increases CFTR chloride transport activity when compared to V470-wildtype CFTR, suppressed the activity of R1235 in such a way that a protein with an open probability not significantly different from V470 wildtype CFTR was obtained. While R628-V CFTR had similar current densities as V470-wildtype CFTR in Xenopus laevis oocytes, R1235-V resulted in current densities that were more than twofold higher than those of V470-wildtype CFTR. However, the current densities generated by R1235/R628-V (R1235 and R628 on a V470 background) CFTR were significant lower than R1235-V or R628-V CFTR. PMID- 10812064 TI - Effect of DNase on the activity of neutrophil elastase, cathepsin G and proteinase 3 in the presence of DNA. AB - It has been shown previously that DNA binds and inhibits neutrophil elastase (NE). Here we demonstrate that DNA has a better affinity for neutrophil cathepsin G (cat G) than for NE and is a better inhibitor of cat G than of NE. DNase generated <0.5 kb DNA fragments inhibit NE and cat G as potently as full length DNA. This rationalises our observation that administration of DNase to cystic fibrosis patients does not enhance the NE and cat G activity of their lung secretions. Neutrophil proteinase 3 is not inhibited by DNA and might thus be the most harmful proteinase in inflammatory lung diseases. PMID- 10812065 TI - Pain peptides. Solution structure of orphanin FQ2. AB - Orphanin FQ2 (OFQ2) is a novel heptadecapeptide generated from prepronociceptin (PPNOC), the same precursor of nociceptin/orphanin FQ and nocistatin. OFQ2 is a potent analgesic when administered both supraspinally and spinally. In order to clarify the structural relationship with all peptides generated from PPNOC, we have undertaken the conformational study of OFQ2 in water and in structure promoting solvent media. Nuclear magnetic resonance data and theoretical calculations are consistent with a well defined helical structure from Met(5) to Ser(16). The uniform distribution of hydrophobic residues along the helix suggests that OFQ2 may interact with the transmembrane helices of a receptor akin to those of nociceptin and opioids. PMID- 10812066 TI - Vascular endothelial growth factor (VEGF) directly enhances osteoclastic bone resorption and survival of mature osteoclasts. AB - In bone development and regeneration, angiogenesis and bone/cartilage resorption are essential processes and are closely associated with each other, suggesting a common mediator for these two biological events. To address this interrelationship, we examined the effect of vascular endothelial growth factor (VEGF), the most critical growth factor for angiogenesis, on osteoclastic bone resorbing activity in a culture of highly purified rabbit mature osteoclasts. VEGF caused a dose- and time-dependent increase in the area of bone resorption pits excavated by the isolated osteoclasts, partially by enhancing the survival of the cells. Two distinct VEGF receptors, KDR/Flk-1 and Flt-1, were detectable in osteoclasts at the gene and protein levels, and VEGF induced tyrosine phosphorylation of proteins in osteoclasts. Thus, osteoclastic function and angiogenesis are up-regulated by a common mediator such as VEGF. PMID- 10812067 TI - Spontaneous galactosylation of agalactoglycoproteins in colostrum. AB - We have found that spontaneous galactosylation of GlcNAc residues occurs in bovine colostrum, but not in dialyzed colostrum, without adding UDP-Gal as a donor substrate. UDP-Gal was shown to be present in bovine colostrum at a level ranging from 200 to 600 microM. When a tracer UDP-[(14)C]Gal was added to the dialyzed colostrum together with a Gal beta1,4-specific beta-galactosidase, remarkable incorporation of radioactivity into 24-28 kDa and 33 kDa RCA1-positive glycoproteins was demonstrated by SDS-PAGE/autoradiography. Some 100-140 kDa agalactoglycoproteins of a CHO mutant cell line were also galactosylated on a blotted membrane by the incubation in the colostrum. PMID- 10812068 TI - Genomic structure and promoter analysis of putative mouse acetyl-CoA transporter gene. AB - The acetyl-CoA transporter gene (Acatn) encodes a hydrophobic, multitransmembrane protein that is involved in the process of O-acetylation of sialic acid residues on gangliosides. O-Acetylated gangliosides have been found to play important roles in tissue development and organization during early embryonic stages. We have cloned the gene for mouse acetyl-CoA transporter. The gene spans approximately 20 kb and is composed of seven exons and six introns. A single transcription initiation site, 371 bp upstream of the ATG start codon, was identified. The promoter region was found to lack a TATA box. However, several potential transcription factor binding motifs such as AP1, AP2, C/EBPalpha, C/EBPbeta, HSF, GATA2 and MZF1 were identified in the promoter region. PMID- 10812069 TI - Hormonal regulation of organic cation transporter OCT2 expression in rat kidney. AB - Rat (r) OCT2 was identified as the second member of the organic cation transporter (OCT) family, and is predominantly expressed in the kidney. We reported previously that rOCT2 was responsible for the gender differences in renal basolateral membrane organic cation transport activity. As renal rOCT2 expression in males is much higher than that in females, we hypothesized that rOCT2 expression may be under the control of sex hormones. Treatment of male and female rats with testosterone significantly increased the expression levels of rOCT2 mRNA and protein in the kidney, whereas estradiol treatment moderately decreased the expression levels of rOCT2. There was no regulation of renal rOCT1 mRNA expression by testosterone or estradiol. Treatment of male and female rats with testosterone significantly stimulated the tetraethylammonium (TEA) accumulation by renal slices, whereas estradiol treatment caused a decrease in the TEA accumulation by slices from male but not female rats. The present findings suggested that testosterone up-regulates renal rOCT2 expression and estradiol moderately down-regulates rOCT2. PMID- 10812070 TI - Catalases and thioredoxin peroxidase protect Saccharomyces cerevisiae against Ca(2+)-induced mitochondrial membrane permeabilization and cell death. AB - The involvement of reactive oxygen species in Ca(2+)-induced mitochondrial membrane permeabilization and cell viability was studied using yeast cells in which the thioredoxin peroxidase (TPx) gene was disrupted and/or catalase was inhibited by 3-amino-1,2, 4-triazole (ATZ) treatment. Wild-type Saccharomyces cerevisiae cells were very resistant to Ca(2+) and inorganic phosphate or t-butyl hydroperoxide-induced mitochondrial membrane permeabilization, but suffered an immediate decrease in mitochondrial membrane potential when treated with Ca(2+) and the dithiol binding reagent phenylarsine oxide. In contrast, S. cerevisiae spheroblasts lacking the TPx gene and/or treated with ATZ suffered a decrease in mitochondrial membrane potential, generated higher amounts of hydrogen peroxide and had decreased viability under these conditions. In all cases, the decrease in mitochondrial membrane potential could be inhibited by ethylene glycol-bis(beta aminoethyl ether) N,N, N',N'-tetraacetic acid, dithiothreitol or ADP, but not by cyclosporin A. We conclude that TPx and catalase act together, maintaining cell viability and protecting S. cerevisiae mitochondria against Ca(2+)-promoted membrane permeabilization, which presents similar characteristics to mammalian permeability transition. PMID- 10812071 TI - The structure of denatured bovine pancreatic trypsin inhibitor (BPTI). AB - In the presence of denaturant and thiol initiator, the native bovine pancreatic trypsin inhibitor (BPTI) denatures by shuffling its native disulfide bonds and converts to a mixture of scrambled isomers. The extent of denaturation is evaluated by the relative yields of the scrambled and native species of BPTI. BPTI is an exceedingly stable molecule and can be effectively denatured only by guanidine thiocyanate (GdmSCN) at concentrations higher than 3-4 M. The denatured BPTI consists of at least eight fractions of scrambled isomers. Their composition varies under increasing concentrations of GdmSCN. In the presence of 6 M GdmSCN, the most predominant fraction of scrambled BPTI accounts for 56% of the total structure of denatured BPTI. Structural analysis reveals that this predominant fraction contains the bead-form isomer of scrambled BPTI, bridged by three pairs of neighboring cysteines, Cys5-Cys14, Cys30-Cys38 and Cys51-Cys55. The extreme conformational stability of BPTI has important implications in its distinctive folding pathway. PMID- 10812073 TI - Two possible conducting states of the influenza A virus M2 ion channel. AB - Molecular dynamics simulations have been performed on protonated four-helix bundles based on the 25-residue Duff-Ashley transmembrane sequence of the M2 channel of the influenza A virus. Well-equilibrated tetrameric channels, with one, two and four of the H37 residues protonated, were investigated. The protonated peptide bundles were immersed in the octane portion of a phase separated water/octane system, which provided a membrane-mimetic environment. The simulations suggest that there could be two conducting states of the M2 channel corresponding to tetramers containing one or two protonated histidines. The more open structure of the doubly protonated state suggests it would have the higher conductance. PMID- 10812072 TI - alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. AB - Voltage-gated K(+) (Kv) channels are particularly important in the physiology of excitable cells in the heart and the brain. PSD-95 is known to cluster Shaker channels and NMDA receptors and the latter is known to couple through alpha actinin-2 to the post-synaptic cytoskeleton [Wyszynski et al. (1997) Nature 385, 439-442], but the mechanisms by which Kv channels are linked to the actin cytoskeleton and clustered at specific sites in the heart are unknown. Here we provide evidence that Kv1.5 channels, widely expressed in the cardiovascular system, bind with alpha-actinin-2. Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with alpha-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. The ion channels and alpha-actinin-2 co-localize at the membrane in HEK cells, where disruption of the actin cytoskeleton and antisense constructs to alpha-actinin-2 modulate the ion and gating current density. PMID- 10812074 TI - Molecular chaperone properties of serum amyloid P component. AB - The selective binding of serum amyloid P component (SAP) to proteins in the pathological amyloid cross-beta fold suggests a possible chaperone role. Here we show that human SAP enhances the refolding yield of denatured lactate dehydrogenase and protects against enzyme inactivation during agitation of dilute solutions. These effects are independent of calcium ions and are not inhibited by compounds that block the amyloid recognition site on the B face of SAP, implicating the A face and/or the edges of the SAP pentamer. We discuss the possibility that the chaperone property of SAP, or its failure, may contribute to the pathogenesis of amyloidosis. PMID- 10812075 TI - Presence of a Na(+)/H(+) exchanger in acidocalcisomes of Leishmania donovani and their alkalization by anti-leishmanial drugs. AB - Acidocalcisomes are acidic vacuoles present in trypanosomatids that contain most of the cellular calcium. The data presented here demonstrate that Leishmania donovani acidocalcisomes possess a Na(+)/H(+) exchanger. 3,5-Dibutyl-4 hydroxytoluene, in the concentration range of 0-20 microM, inhibited the Na(+)/H(+) exchanger, and strongly stimulated the activity of the vacuolar H(+) ATPase responsible for vacuolar acidification. As occurs with Na(+), the cationic anti-leishmanial drugs pentamidine, WR-6026, and chloroquine promoted a fast and extensive alkalization of the L. donovani acidocalcisomes. PMID- 10812076 TI - Proadrenomedullin N-terminal 20 peptide (PAMP) elevates blood glucose levels via bombesin receptor in mice. AB - We found a potent hyperglycemic effect of proadrenomedullin N-terminal 20 peptide (PAMP) after intra-third cerebroventricular administration at a dose of 10 nmol in fasted mice. PAMP has four homologous residues with bombesin (BN), a hyperglycemic peptide. PAMP showed affinity for gastrin-releasing peptide preferring receptor (GRP-R) and neuromedin B preferring receptor. The PAMP induced hyperglycemic effect was inhibited by [D-Phe(6), Leu-NHEt(13), des Met(14)]-BN (6-14), GRP-R specific antagonist, indicating that the hyperglycemic effect is mediated at least in part via GRP-R. Furthermore, pretreatment of alpha adrenergic blocker inhibited the PAMP-induced hyperglycemia and hyperglucagonemia, suggesting that the increase of glucagon secretion through alpha-adrenergic activation is involved in this hyperglycemic effect of PAMP. PMID- 10812077 TI - Ribonucleotide reductase is regulated via the R2 subunit during the life cycle of Trypanosoma brucei. AB - We have examined the occurrence of the R1 and R2 subunits of ribonucleotide reductase during the life cycle of Trypanosoma brucei. Whereas the R1 protein is present throughout the life cycle, the R2 protein is not found in cell cycle arrested short stumpy trypanosomes. RT-PCR/hybridization analysis revealed almost equal amounts of the R1 and R2 mRNAs in all life cycle stages of the parasite. The data indicate that ribonucleotide reductase of African trypanosomes is developmentally controlled by post-transcriptional regulation of the R2 subunit. PMID- 10812078 TI - A membrane-permeant ester of phosphatidylinositol 3,4, 5-trisphosphate (PIP(3)) is an activator of human neutrophil migration. AB - Activity of phosphatidylinositol (PI) 3-kinase is required for optimal migration of human neutrophils [Niggli and Keller (1999) Eur. J. Pharmacol. 335, 43-52]. We have tested the direct effect of a product of PI 3-kinase, phosphatidylinositol 3,4,5-trisphosphate (PIP(3)), on neutrophil migration. To this end, a membrane permeant ester of PIP(3), dilauroyl phosphatidylinositol 3,4, 5-trisphosphate heptakis-(acetooxymethyl)ester (PIP(3)/AM) was used. PIP(3)/AM (ED(50): 10-17 microM) induced development of polarity and accumulation of F-actin in the leading lamellae in up to 70% of the cells. These cells exhibited stimulated random migration, comparable to that observed in uniform concentrations of chemotactic peptide. Evidence is provided for a role of Rho-kinase and for activation of PI 3-kinase in a positive feedback loop in PIP(3)/AM-induced motility. PMID- 10812079 TI - Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface: use of immunostaining technique with monoclonal antibodies specific for PI 3,4-P(2). AB - Phosphatidylinositol (PI) 3,4-P(2) is a phosphoinositide that has been shown to be important for signal transduction in growth factor stimulation. We have produced monoclonal antibodies specific for PI 3,4-P(2), which were able to detect PI 3,4-P(2) generated in 293T cells treated with H(2)O(2), or in MKN45/BD110 cells expressing activated PI 3-kinase in immunostaining. Prolonged treatment with 0.05% Tween 20 resulted in detection of staining not only at the plasma membrane, but also at the nuclear surface, indicating that 3' phosphorylated phosphoinositides can be generated and function in the nucleus. PMID- 10812080 TI - Enhanced toxicity of Bacillus thuringiensis Cry3A delta-endotoxin in coleopterans by mutagenesis in a receptor binding loop. AB - We used site-directed mutagenesis to modify the Bacillus thuringiensis cry3A gene in amino acid residues 350-354. Two mutant toxins, A1 (R(345)A,Y(350)F,Y(351)F) and A2 (R(345)A,DeltaY(350), DeltaY(351)), showed significantly improved toxicity against Tenebrio molitor (yellow mealworm). The mutant toxin A1 was also more potent against both Leptinotarsa decemlineata (Colorado potato beetle) and Chrysomela scripta (cottonwood leaf beetle), while A2 displayed enhanced toxicity only in L. decemlineata. Competitive binding assays of L. decemlineata brush border membrane vesicles (BBMV) revealed that binding affinities for the A1 and A2 mutant toxins were ca. 2.5-fold higher than for the wild-type Cry3 toxin. Similar binding assays with C. scripta BBMV revealed a ca. 5-fold lower dissociation rate for the A1 mutant as compared to that of Cry3A. PMID- 10812081 TI - A novel human processed gene, DAD-R, maps to 12p12 and is expressed in several organs. AB - A cDNA of a processed gene of human DAD-1 (defender against apoptotic cell death) was cloned from the human neuroblastoma cell line SH-SY5Y. The genomic sequence of this novel processed gene, DAD-R, lacked introns and was flanked by 8 bp terminal repeats. RT-PCR showed that the transcript is expressed predominantly in testis, ovaries, pancreas, lung and skeletal muscle. DAD-R has several possible initiation codons, one of them producing an open reading frame comprising 75% of the DAD-1 gene. We determined the chromosomal localization of DAD-R as 12p11.2 12p12.1, an area linked to familial synpolydactyly and frequently amplified in a variety of cancers, including those of testis, ovaries, pancreas and lungs. PMID- 10812082 TI - Sweet is stable: glycosylation stabilizes collagen. AB - For most collagens, the melting temperature (T(m)) of the triple-helical structure of collagen correlates with the total content of proline (Pro) and 4 trans-hydroxyproline (Hyp) in the Xaa and Yaa positions of the -Gly-Xaa-Yaa- triplet repeat. The cuticle collagen of the deep-sea hydrothermal vent worm Riftia pachyptila, despite a very low content of Pro and Hyp, has a relatively high thermal stability. Rather than Hyp occupying the Yaa position, as is normally found in mammalian collagens, this position is occupied by threonine (Thr) which is O-glycosylated. We compare the triple-helix forming propensities in water of two model peptides, Ac-(Gly-Pro-Thr)(10)-NH(2) and Ac-(Gly-Pro Thr(Galbeta))(10)-NH(2), and show that a collagen triple-helix structure is only achieved after glycosylation of Thr. Thus, we show for the first time that glycosylation is required for the formation of a stable tertiary structure and that this modification represents an alternative way of stabilizing the collagen triple-helix that is independent of the presence of Hyp. PMID- 10812083 TI - Inhibition of telomerase activity by a cell-penetrating peptide nucleic acid construct in human melanoma cells. AB - We investigated the effect of two peptide nucleic acids (PNAs), which are complementary to the RNA component of human telomerase, on the catalytic activity of the enzyme. PNAs induced a dose-dependent reduction of telomerase activity in cell extracts from human melanoma cell lines and surgical specimens. To down regulate telomerase in intact cells, we generated a chimeric molecule synthesized by coupling the 13-mer PNA to the Antennapedia peptide. The PNA construct induced a dose- and time-dependent inhibition of telomerase activity. However, a 20-day exposure to the PNA construct only caused a slight increase in melanoma cell doubling time and failed to induce any telomere shortening. PMID- 10812084 TI - Detection of lipofuscin-like fluorophore in oxidized human low-density lipoprotein. 4-hydroxy-2-nonenal as a potential source of fluorescent chromophore. AB - It has recently been shown that the lipid peroxidation product 4-hydroxy-2 nonenal (HNE) forms a fluorescent hydroxyiminodihydropyrrole derivative with the epsilon-amino group of lysine residue. In this study, we raised a monoclonal antibody (mAb2C12) directed to the fluorophore-protein conjugate and found that the antibody was specific to the chromophore structure of the compound. Immunohistochemical analysis of atherosclerotic lesions from the human aorta showed that the fluorophore was indeed present in the lesions, in which intense positivity was primarily associated with macrophage-derived foam cells and thickening of the neointima of the arterial walls. Antigenic materials were also detected in the oxidatively modified low-density lipoprotein (LDL) with Cu(2+) and in the oxidatively modified bovine serum albumin with an iron/linoleic acid autoxidation system, indicating that the HNE, which originated from the peroxidation of polyunsaturated fatty acids, could be a potential source of the fluorescent chromophore in oxidized LDL. PMID- 10812085 TI - Dissection of the maturation reactions of the [NiFe] hydrogenase 3 from Escherichia coli taking place after nickel incorporation. AB - The steps in the maturation of the precursor of the large subunit (pre-HycE) of hydrogenase 3 from Escherichia coli taking place after incorporation of both iron and nickel were investigated. Pre-HycE could be matured and processed in the absence of the small subunit but association with the cytoplasmic membrane required heterodimer formation between the two subunits. Pre-HycE formed a complex with the chaperone-like protein HypC in the absence of the small subunit and, in this complex, also incorporated nickel. For the C-terminal processing, HypC had to leave the complex since only a HypC-free, nickel-containing form of pre-HycE was a substrate for the maturation endopeptidase. PMID- 10812086 TI - FTIR spectroscopy of complexes formed between metarhodopsin II and C-terminal peptides from the G-protein alpha- and gamma-subunits. AB - Metarhodopsin II (MII) provides the active conformation of rhodopsin for interaction with the G-protein, Gt. Fourier transform infrared spectra from samples prepared by centrifugation reflect the pH dependent equilibrium between MII and inactive metarhodopsin I. C-terminal synthetic peptides (Gtalpha(340-350) and Gtgamma(60-71)farnesyl) stabilize MII. We find that both peptides cause similar spectral changes not seen with control peptides (Gtalpha (K341R, L349A) and non-farnesylated Gtgamma). The spectra reflect all the protonation dependent bands normally observed when MII is formed at acidic pH. Beside the protonation dependent bands, additional features, similar with both peptides, appear in the amide I and II regions. PMID- 10812088 TI - Presidential address PMID- 10812089 TI - Address at opening ceremony PMID- 10812087 TI - Inhibition of voltage-dependent sodium channels by Ro 31-8220, a 'specific' protein kinase C inhibitor. AB - We find that several protein kinase C (PKC) inhibitors, previously considered to be specific, directly inhibit voltage-dependent Na(+) channels at their useful concentrations. Bisindolylmaleimide I (GF 1092037), IX (Ro 31-8220) and V (an inactive analogue), but not H7 (a non-selective isoquinolinesulfonamide protein kinase inhibitor), inhibited Na(+) channels assessed by several independent criteria: Na(+) channel-dependent glutamate release and [(3)H]batrachotoxinin-A 20-alpha-benzoate binding in rat cortical synaptosomes, veratridine-stimulated 22Na(+) influx in CHO cells expressing rat CNaIIa Na(+) channels and Na(+) currents measured in isolated rat dorsal root ganglion neurons by whole cell patch-clamp recording. These findings limit the usefulness of the bisindolylmaleimide class PKC inhibitors in excitable cells. PMID- 10812090 TI - Acquired tolerance, allograft "acceptance," and immune suppression. PMID- 10812091 TI - Long-term survival after orthotopic liver transplantation with regard to country of origin and residence. PMID- 10812092 TI - Changes in serum concentrations of matrix metalloproteinases in kidney transplantation. PMID- 10812093 TI - Emerging therapeutic strategies for the prevention and treatment of chronic allograft rejection. PMID- 10812094 TI - Renal transplantation in children. PMID- 10812095 TI - Higher immunosuppressive efficacy of mycophenolate mofetil in combination with FK 506 than in combination with cyclosporine A. PMID- 10812096 TI - Evaluation of endourological interventions used to treat urological complications in 394 kidney recipients. PMID- 10812097 TI - Pretransplantation human herpes virus 8 seropositivity as a risk factor for Kaposi's sarcoma in kidney transplant recipients. PMID- 10812098 TI - Histologic evaluation, HLA-DR expression, and macrophage density of renal biopsies in OKT3-treated acute rejection: comparison with steroid response in acute rejection. PMID- 10812099 TI - Clinical trial issues in studies of chronic rejection. PMID- 10812100 TI - Aneurysms of the hepatic artery after liver transplantation. PMID- 10812101 TI - Heterotopic segmental liver transplantation in children. PMID- 10812102 TI - Infectious complications after orthotopic liver transplantation with different immunosuppressive induction regimens. PMID- 10812103 TI - Caval complications after orthotopic liver transplantation. PMID- 10812104 TI - Adverse effects of a controlled living-unrelated donor renal transplant program on living-related and cadaveric kidney donation. PMID- 10812105 TI - Left ventricular hypertrophy and angiotensin-converting enzyme gene polymorphism in renal allograft recipients. PMID- 10812106 TI - Estimation of glomerular filtration rate in renal transplants based on serum creatinine level after oral cimetidine. PMID- 10812107 TI - Clinical identification of transplant antigens. PMID- 10812108 TI - Antithrombotic effects of aspirin after renal transplantation. PMID- 10812109 TI - Increased incidence of infection in verapamil-treated kidney transplant recipients. PMID- 10812110 TI - Absence of Epstein-Barr virus DNA sequences in iatrogenic Kaposi's sarcomas of renal transplant recipients. PMID- 10812111 TI - Efficiency of preventive treatment for osteoporosis after renal transplantation. PMID- 10812113 TI - Impact of HCV infection on development of posttransplantation diabetes mellitus in renal allograft recipients. PMID- 10812112 TI - Histologic outcome of hepatitis C virus infection in renal transplant recipients and the effect of pretransplantation interferon treatment. PMID- 10812114 TI - Opportunistic pulmonary infection after renal transplantation. PMID- 10812115 TI - Patient blood glucose levels before and after kidney transplantation. PMID- 10812116 TI - Effects of hepatitis C virus infection on cyclosporine trough levels in renal transplant patients. PMID- 10812117 TI - One center's experience with OKT3 in renal transplantation: patient and graft survival, complications, and cost analysis. PMID- 10812118 TI - Role of immunonutrition in reducing complications following organ transplantation. PMID- 10812119 TI - Inflammatory vascular response and microcirculatory hemodynamics during acute rejection phase in composite tissue allografts. PMID- 10812120 TI - Ureteral complications in 1100 consecutive renal transplants. PMID- 10812121 TI - Pretransplantation alpha-interferon therapy and the effect of hepatitis C virus infection on kidney allograft recipients. PMID- 10812122 TI - Heart transplantation after mechanical circulatory support. PMID- 10812123 TI - Lymphoma-the most common neoplasia in renal transplant recipients. PMID- 10812124 TI - Donor hospital development in Germany. PMID- 10812125 TI - Effect of low-dose donor radiation on acute rejection of composite limb allografts. PMID- 10812126 TI - Comparison of the intraoperative transfusion requirements and arterial blood gases in hetero-versus orthotopic liver transplantation. PMID- 10812127 TI - Xenograft and homograft transplantation in cardiac surgery at Baskent University Hospital. PMID- 10812128 TI - A new regimen of MESNA (2-mercaptoethanesulfonate) effectively prevents cyclophosphamide-induced hemorrhagic cystitis in bone marrow transplant recipients. PMID- 10812129 TI - Results of allogeneic bone marrow transplantation in 90 patients at a University Hospital in Southern Iran. PMID- 10812130 TI - Evaluation of HLA antibody synthesis due to blood transfusion in 314 patients from the Isfahan (Iran) Transplantation Center. PMID- 10812131 TI - Long-term survival and complications after orthotopic liver transplantation for postnecrotic cirrhosis. PMID- 10812132 TI - Randomized trial comparing neoral and tacrolimus immunousuppression for recipients of renal transplants procured from different donor groups. PMID- 10812133 TI - Volume measurement by computed tomography in auxiliary heterotopic partial liver transplant recipients: follow-up results. PMID- 10812134 TI - Double renal arteries in living-related kidney transplantation. PMID- 10812135 TI - Renal transplantation in patients with amyloidosis. PMID- 10812136 TI - Factors influencing response to hepatitis B virus vaccination in hemodialysis patients. PMID- 10812138 TI - Corrective surgery for high-output brachial artery-basilic vein arteriovenous fistulae: a new approach. PMID- 10812137 TI - Endoscopic treatment of vesicoureteral reflux in renal transplant candidates. PMID- 10812139 TI - Patient-controlled epidural analgesia with morphine in renal transplant patients. PMID- 10812140 TI - Acute rejection in protocol renal transplant biopsies-institutional variations. PMID- 10812141 TI - Multiple-artery anastomosis in kidney transplantation. PMID- 10812142 TI - New immunosuppressive agents. PMID- 10812143 TI - Comparison of the effects of simvastatin and pravastatin on acute rejection episodes in renal transplant patients. PMID- 10812144 TI - Results of treatment in renal transplant patients with Kaposi sarcoma: one-center experience. PMID- 10812145 TI - Attitudes of religious people in Turkey regarding organ donation and transplantation. PMID- 10812146 TI - Renal dysfunction in orthotopic liver transplantation using simple cross-clamping technique and no venovenous bypass. PMID- 10812147 TI - Pediatric renal transplantation. PMID- 10812148 TI - Use of antifibrinolytics in orthotopic liver transplantation. PMID- 10812149 TI - On the failure of health reforms. PMID- 10812150 TI - Noninvasive heart monitoring after heart transplantation with CHARM (computerized heart allograft recipient monitoring): clinical experience. PMID- 10812151 TI - Factors influencing organ donation and transplantation in the Middle East. PMID- 10812152 TI - Factors influencing long-term graft loss. The Collaborative Transplant Study. PMID- 10812153 TI - The challenge of posttransplant tuberculosis. PMID- 10812154 TI - Pediatric renal transplantation in Pakistan. PMID- 10812155 TI - Use of an alloreactive lymphocyte vaccine to prolong rat cardiac allograft survival. PMID- 10812156 TI - Drug-drug interaction between chloramphenicol and tacrolimus in a liver transplant recipient. PMID- 10812157 TI - Renal retransplantation in elderly recipients under tacrolimus-based immunosuppression. PMID- 10812158 TI - Use of phenobarbital in the management of acute tacrolimus toxicity: a case report. PMID- 10812160 TI - Advances in the vascular pathophysiology of ischemic stroke. AB - The cerebral vascular supply is constructed to protect the cerebral hemispheres and brainstem from the consequences of blood flow cessation. Reversal of blood flow around local obstructions is a feature of the microvascular beds of the striatum and cerebral cortex. Cerebral capillaries of these beds consist of endothelial cells, basal lamina, and astrocyte end-feet that sit in close apposition. The interaction of astrocytes with neurons indicates the close relationship of microvessels to neurons. These relationships are altered when blood flow ceases in the supplying artery. Increased endothelial cell permeability and endocytoses lead to edema formation, and matrix degradation is associated with hemorrhage. Autoregulation is lost. Ischemia initiates leukocyte adhesion receptor expression, which is promoted by cytokine generation from the neuropil and activated monocytes. "Preactivation" may further augment the inflammatory responses to ischemia. The activation of cerebral microvessels by ischemia is heterogeneous, involving alterations in integrin-matrix interactions, leukocyte-endothelial cell adhesion, permeability changes, and the "no-reflow" phenomenon due to platelet activation, fibrin formation, and leukocyte adhesion. Ischemia produces swelling of the microvascular endothelium, and rapid detachment and swelling of the astrocyte end-feet. Ischemic injury targets the microvasculature, where the inflammatory responses are initiated and contribute to tissue injury. PMID- 10812161 TI - Long-term prevention of ischaemic stroke and stroke recurrence. AB - Stroke is the third most important cause of mortality, but the leading cause of severe handicap, dependency, and loss of social competence. Because of the high recurrence rate, active secondary prevention is mandatory once a stroke has occurred. Secondary prevention of stroke implies the primary prevention of cardiovascular disorders as well. Among the modifiable risk factors hypertension is worst and should be normalized according to recent WHO criteria, also in the elderly. Smoking is another major risk factor and hard to delete. Diabetes mellitus and hyperlipidaemia are also important risk factors and should be treated consequently by diet and medication. Moderate alcohol intake, normalization of body weight and regular physical activity also contribute considerably to prevention of stroke. Whether hyperhomocysteinaemia should be normalized has not yet been clarified. Cardiovascular disorders are an important source of ischemic strokes, particularly atrial fibrillation. Low dose anticoagulation can dramatically reduce stroke risk. Carotid endarterectomy in symptomatic stenoses is the most expensive means of stroke prevention. In less severe stenoses, or ICA occlusions, antiplatelet agents are the treatment of choice. Composite drugs with ASS and other antiplatelet agents seem to be superior to either compound alone. Dissections of the cervical arteries should not be operated on but may be treated by anticoagulation or antiplatelet agents in the acute and subacute phase. The potency of a consequent and comprehensive stroke prevention in preventing disability and death is much greater than any sophisticated acute stroke treatment. PMID- 10812162 TI - Acute ischemic stroke management. AB - The use of thrombolytic therapy represents one of many recent developments in the management of acute ischemic stroke. The development of stroke teams and protocols has been driven by these new demands for an urgent response to ischemic stroke. The short time window of 3 hours for therapy with intravenous recombinant tissue plasminogen activator requires efficient evaluation and treatment of stroke patients and also necessitates a rigorous approach to blood pressure management, electrolytes, fluids, and temperature. Anticoagulation has not been proven to safely prevent progression or early recurrence of stroke, but antiplatelet therapy is worthwhile. PMID- 10812163 TI - Mutation of the RB1 gene caused unilateral retinoblastoma in early Age. AB - Fluorescence in situ hybridization (FISH) was applied for the detection of the retinoblastoma tumor-suppressor gene deletion on retinoblastoma tumor cells obtained from the unilateral tumor of a 3-month-old boy. Both retinoblastoma tumor cells and peripheral lymphocytes of the patient showed one hybridization signal per cell at the retinoblastoma-1 locus, indicating that one copy of the gene was deleted. Peripheral blood lymphocytes obtained from the patient's parents had two copies per cell for the gene. Retinoblastoma nuclear phosphoprotein expression could not be detected in the tumor tissue. No copy number alterations were detected with ten different centromeric DNA probes in the tumor cells. The deletion at the RB1 locus detected by FISH suggested that this gene alteration was heritable. The parental peripheral blood lymphocytes did not show the loss of the gene; thus the first deletion may have taken place in either of the parental germ cells. The second somatic mutation of the RB1 gene was probably under the detection limit of FISH. The second allelic alterations were detected by using the polymerase chain reaction for all exons of the retinoblastoma gene. PMID- 10812164 TI - Transposition of duplicated chromosomal segment involving fused BCR-ABL gene or ABL oncogene alone in chronic myelocytic leukemia and Ph chromosome-positive acute leukemia with complex karyotypes. AB - Thirty-six patients with chronic myelocytic leukemia (CML) in the blastic phase were examined by fluorescence in situ hybridization to clarify the mechanisms of progression of the disease. Two of 19 CML patients in the blastic phase (10.5%) had an extra fused BCR-ABL gene on structurally complex chromosome aberrations in addition to the Ph chromosome. Another patient had an extra ABL oncogene on the end of a deleted chromosome, resulting in three copies of the ABL oncogene. These three patients showed additional chromosome aberrations, such as der(12), der(15), and der(18), which differ from the standard karyotypic evolution in the blastic phase. Amplification of the fused BCR-ABL gene or the ABL oncogene seemed to be induced by transposition. These segmental transpositions suggest that these regions have high genetic instability possibly leading to blastic transformation. PMID- 10812165 TI - Precise localization by microdissection/reverse ISH and FISH of the t(15;17)(q24;q21.1) chromosomal breakpoints associated with acute promyelocytic leukemia. AB - The acute promyelocytic leukemia (APL M3)-associated translocation (15;17) has been described as having breakpoints variably located between 15q22 and 15q26, and 17q11 and 17q25. Most of the recent studies using DNA probes (fluorescence in situ hybridization [FISH]) for analysis have indicated the chromosome 15 breakpoint to be in 15q22. We have utilized a combination of G-banding, FISH, and chromosome microdissection/reverse ISH to precisely map the breakpoint to t(15;17)(q24;q21.1). PMID- 10812166 TI - Characterization of chromosomal aberrations in human gastric carcinoma cell lines using chromosome painting. AB - Using chromosome painting, a study of chromosomal abnormalities was performed in six gastric carcinoma cell lines (SNU-484, 601, 620, 638, 668, 719) from Korean patients. Each carcinoma cell line had unique modal karyotypic characteristics and showed a variable number of numerical and structural clonal cytogenetic aberrations. SNU-484, SNU-620, and SNU-668 had near-triploidy; SNU-601, SNU-638, and SNU-719 had near-diploidy. The origins of the marker chromosomes of these cell lines were identified by fluorescence in situ hybridization with constructed painting probes. In all of six cell lines, rearrangement of chromosome 17 resulting in partial deletion of 17p (and/or partial duplication of 17q) was found. The most frequent marker was a partial gain of chromosome 7 with the breakpoints on 7q22 and 7q31. The nonrandom rearrangements of chromosomes were also determined on 1q32, 5q11-q22, 8q, 14q22, 14q34, and 15q15; suggesting that they may be the candidate regions for the isolation of the genes related to gastric cancer. PMID- 10812167 TI - Trisomy 4 leading to duplication of a mutated KIT allele in acute myeloid leukemia with mast cell involvement. AB - A G-->T transversion at nucleotide 2467 of the c-KIT gene leading to Asp816-->Tyr (D816Y) substitution in the phosphotransferase domain has been previously identified in a patient with rapidly progressing AML-M2 and mast cell involvement; the patient's blasts had a 47,XY, +4,t(8;21)(q22;q22) karyotype. Herein we confirm the simultaneous presence of both major chromosomal changes by multicolor fluorescence in situ hybridization (FISH) on interphase CD34+ mononuclear cells. By setting up culture leukemic blasts, spontaneous differentiation of adherent cells with mast-cell like features was proved by histochemical and immunoenzymatic analyses. Fluorescence in situ hybridization evidence of trisomy 4 confirmed the origin of differentiated cells from the leukemic blasts. Semiquantitative polymerase chain reaction (PCR) and phosphoimage densitometry of wild-type and mutated KIT alleles on bone marrow blasts made it possible to demonstrate that chromosome 4 trisomy led to a double dosage of the mutated KIT allele. This finding, and that of trisomy 7 and MET mutation in hereditary renal carcinoma represent the only cases of human tumors in which an increased number of chromosomes carrying an oncogene activated by point mutation have been detected. PMID- 10812168 TI - Cytogenetic characterization of childhood hepatoblastoma. AB - We describe the cytogenetic abnormalities in two cases of childhood hepatoblastoma. The first case was of fetal histology with squamous metaplasia, and cytogenetic study showed an add(5)(q31). Although an association between hepatoblastoma and familial adenomatous polyposis is recognized, the breakpoint in this case is distal to 5q21 and most probably does not involve the APC gene at that location. The second case was of macrotrabecular histology, and cytogenetic study showed an unbalanced translocation in the form of der(4)t(1;4)(q12;q34) in a hyperdiploid clone. Including our case, der(4)t(1;4)(q12;q34) has been recognized in four cases of hepatoblastoma, and it may be the first recurrent translocation in this tumor. Understanding the molecular mechanism and clinical significance of this translocation awaits analysis of more cases. PMID- 10812169 TI - A case of papillary meningioma with a t(1;4)(q44;q21). AB - We report the results of cytogenetic analyses of three cases of meningiomas. The first case, a papillary meningioma, showed only one cytogenetic abnormality, 46,XX,t(1;4)(q44;q21). In contrast, the other two benign fibroblastic meningiomas showed loss of chromosome 22. Loss and/or rearrangement of chromosomes other than chromosome 22 appears to be associated with a more aggressive clinical course. It is suggested that a sole cytogenetic abnormality with a normal chromosome 22 indicates an atypical nature of meningioma. PMID- 10812170 TI - PTEN mutation analysis in two genetic subtypes of high-grade oligodendroglial tumors. PTEN is only occasionally mutated in one of the two genetic subtypes. AB - We recently identified two genetic subtypes of high-grade oligodendroglial tumors (HG-OT): 1p-/19q- HG-OT are characterized by a loss of chromosome 1p32-36 (del(1)(p32-p36) and/or a del(19)(q13. 3); whereas +7/-10 HG-OT harbor a gain of chromosome 7 (+7) and/or a -10 without a loss of 1p32-36 and 19q13.3. Because a 10 and a +7 are most frequently detected in glioblastomas (GBM), the genotype of +7/-10 HG-OT suggests that these tumors are GBM with a prominent oligodendroglial phenotype rather than anaplastic oligodendrogliomas. PTEN is a tumor suppressor gene, located at 10q23.3, which is involved in tumor progression of GBM and other neoplasms. In this study, we screened for PTEN mutations in six low-grade oligodendroglial tumors (LG-OT), five 1p-/19q- HG-OT, seven +7/-10 HG-OT, and nine xenografted GBM. PTEN mutations were detected in none of the LG-OT and 1p /19q- HG-OT, once in +7/-10 HG-OT, and frequently in GBM. As one of the +7/-10 HG OT harbored a PTEN mutation, this demonstrates that PTEN can be involved in the oncogenesis of this genetic subtype of HG-OT. The lower frequency of PTEN mutations in +7/-10 HG-OT compared to GBM suggests that these tumors are of a distinct tumor type rather than GBM. Published by Elsevier Science Inc. PMID- 10812171 TI - Comparative genomic hybridization analysis detects frequent over-representation of DNA sequences at 3q, 7p, and 8q in head and neck carcinomas. AB - Comparative genomic hybridization (CGH) was used to identify chromosomal imbalances in 19 samples of squamous cell carcinoma of the head and neck (HNSCC). The chromosome arms most often over-represented were 3q (48%), 8q (42%), and 7p (32%); in many cases, these changes were observed at high copy number. Other commonly over-represented sites were 1q, 2q, 6p, 6q, and 18q. The most frequently under-represented segments were 3p and 22q. Loss of heterozygosity of two polymorphic microsatellite loci from chromosome 22 was observed in two tongue tumors, in agreement with the CGH analysis. Gains of 1q and 2q material were detected in patients exhibiting a clinical history of recurrence and/or metastasis followed by terminal disease. This association suggests that gain of 1q and 2q may be a new marker of head and neck tumors with a refractory clinical response. PMID- 10812172 TI - Anomalous transcripts and allelic deletions of the FHIT gene in human esophageal cancer. AB - The fragile histidine triad (FHIT) gene is localized on chromosome 3p14 and spans the common fragile site FRA3B. Even though its role in carcinogenesis is still unclear, this gene is frequently inactivated by carcinogen-induced intragenic deletions in many types of cancers, and FHIT abnormal transcripts are found in many primary tumors and tumor-derived cell lines. We evaluated FHIT gene involvement in 39 esophageal carcinomas (18 adenocarcinomas [AC?, 21 squamous cell carcinomas [SCC]) by both reverse transcriptase-polymerase chain reaction (RT-PCR) amplification and loss of heterozygosity analysis (LOH). Thirty cases (77%) displayed either aberrant FHIT transcripts (12 cases) and/or LOH (24 cases); among these, only 6 samples displayed both aberrant transcripts and LOH, thus suggesting that the two events are probably independent. Moreover, LOH was significantly higher in SCC (80%) than in AC (44%), and because most of our patients are heavy smokers and/or alcohol consumers, these results suggest that the FHIT gene might be a common target for carcinogens also in the esophagus. PMID- 10812173 TI - Cytogenetics of epithelial hyperplasias of the human breast. AB - Generally, benign breast lesions behave like innocuous and limited proliferations; however, sometimes they can represent precancerous pathologies. The cytogenetic analysis of five mammary epithelial hyperplasias is reported. Four cases had clonal chromosome alterations. All of the cases presented a modal number of 46 chromosomes. Chromosome 9 monosomies and chromosome 1 deletions were common in these benign tumors. The study of benign proliferations of the breast may reveal a possible relationship between chromosomal alterations and the conditions of the tissue. PMID- 10812174 TI - Spurious monosomy 7 in leukemia due to centromeric heteromorphism. AB - We report heteromorphism of the centrometric region of human chromosome 7, which was observed in the cytogenetic assessment of a complete remission of a pre-B acute lymphoblastic leukemia in the bone marrow cells of a 25-year-old woman. Classical cytogenetic study was performed, as well as metaphase and interphase fluorescence in situ hybridization (FISH) carried out with an alphoid DNA probe specific for the chromosome 7 centromere for detection of leukemic clones with monosomy 7 found at the initial diagnosis. We show an important centromeric heteromorphism of this chromosome detected by FISH and clearly visible on all metaphases and nuclei analyzed. This heteromorphism is observable as a fluorescent signal five- or sixfold larger than that on the homologue. To our knowledge, this heteromorphism of chromosome 7 has not been reported in the literature. However, with the use of FISH analysis, it could be easily mistaken for a mosaicism of monosomy 7, which can be misleading in the interpretation of the results. PMID- 10812175 TI - A 12q13 translocation involving the HMGI-C gene in richter transformation of a chronic lymphocytic leukemia. AB - We report a case of Richter transformation of a chronic lymphocytic leukemia with a 12q13 translocation involving the HMGI-C gene. Fluorescence in situ hybridization analysis with the use of two different cosmid pools spanning the entire HMGI-C region showed that the breakpoint on chromosome 12 was located in the HMGI-C gene, presumably within intron 3. In fact, the 3' region of HMGI-C had been translocated to a derivative chromosome 6. This translocation was not visible at the cytogenetic level. Immunohistochemical analysis performed on the bone marrow smear demonstrated the expression of the HMGI-C protein specifically in the blasts, suggesting that the aberrant expression of the HMGI-C gene might have an important role in the process of leukemogenesis. PMID- 10812176 TI - Duplication of 1q in a child with down syndrome and myelodysplastic syndrome. AB - Cytogenetic analysis of bone marrow cells was performed on a 2-year-old African American male with Down syndrome (DS) and myelodysplastic syndrome (MDS), specifically refractory anemia with excess blasts in transformation (RAEB-T). Chromosome analysis showed, in addition to the constitutional trisomy 21, a trisomy of chromosome 11 and a dup(1)(q23q31). This duplication of 1q is apparently a new chromosomal abnormality in a child with MDS. Partial trisomy of the long arm of chromosome 1 has been reported by several authors and appears to represent a nonrandom chromosomal anomaly in patients with MDS/acute myelogenous leukemia and DS. PMID- 10812177 TI - A novel translocation (17;19)(p13;p13) in a patient with acute myelomonocytic leukemia. AB - We report on a patient with acute myeloid leukemia (AML M4) and a so far unrecorded translocation (17;19). The leukemia transformed from a myeloproliferative disorder (MPD) and showed a progressive fatal course. Following transformation, all leukemic cells showed an apparently balanced translocation (17;19)(p13;p13). The breakpoint regions harbor genes such as TP53 (17p13) and E2A, ENL, or LYL1 (19p13), which could be relevant in leukemogenesis. We suspect that the translocation (17;19)(p13;p13) may be a prognostic factor for transformation from chronic MPD to acute leukemia. PMID- 10812178 TI - An isochromosome 6p in a primary meningeal malignant melanoma. AB - The rearrangement of chromosome 6, particularly the deletion of 6q, has been observed in human malignant melanoma with or without brain metastases. The isochromosome 6p has also been described. In this study, we report the cytogenetic analysis of a primary malignant melanoma of the central nervous system. Its dominating karyotype was 47,XX,+i(6)(p10). Fluorescence in situ hybridization (FISH), using a 6p chromosome arm probe, confirmed the structure of the isochromosome. To our knowledge, this is the first report of this type of chromosomal aberration in an uncommon neoplasm of leptomeningeal melanocytic origin. PMID- 10812179 TI - Development of a mixed mode adsorption process for the direct product sequestration of an extracellular protease from microbial batch cultures. AB - Direct product sequestration of extracellular proteins from microbial batch cultures can be achieved by continuous or intermittent broth recycle through an external extractive loop. Here, we describe the development of a fluidisable, mixed mode adsorbent, designed to tolerate increasing ionic strength (synonymous with extended productive batch cultures). This facilitated operations for the integrated recovery of an extracellular acid protease from cultures of Yarrowia lipolytica. Mixed mode adsorbents were prepared using chemistries containing hydrophobic and ionic groups. Matrix hydrophobicity and titration ranges were matched to the requirements of integrated protease adsorption. A single expanded bed was able to service the productive phase of growth without recourse to the pH adjustment of the broth previously required for ion exchange adsorption. This resulted in increased yields of product, accompanied by further increases in enzyme specific activity. A step change from pH 4.5 to 2.6, across the isoelectric point of the protease, enabled high resolution fixed bed elution induced by electrostatic repulsion. The generic application of mixed mode chemistries, which combine the physical robustness of ion-exchange ligands in sanitisation and sterilisation procedures with a selectivity, which approaches that of affinity interactions, is discussed. PMID- 10812180 TI - Isolation of mesophilic solvent-producing clostridia from Colombian sources: physiological characterization, solvent production and polysaccharide hydrolysis. AB - One hundred and seventy-eight new butanol-acetone producing bacteria related to saccharolytic clostridia were isolated from agricultural sources in Colombia and their fermentation potential was evaluated. Thirteen isolates produced more total solvents from glucose than Clostridium acetobutylicum ATCC 824. The isolates with the highest single solvent production were IBUN 125C and IBUN 18A with 0.46 mol butanol and 0.96 mol ethanol formed from 1 mol glucose, yielding 25. 2 and 29.1 g l(-1) total solvents, respectively, which is close to the maximum values described to date. Most of the new isolates produced exoenzymes for the hydrolysis of starch, carboxymethyl cellulose, xylan, polygalacturonic acid, inulin and chitosan. Together with the high efficiency of solvent production, these hydrolytic isolates may be useful for the direct fermentation of biomass. According to their physiological profile, the most solvent-productive isolates could be classified as strains of C. acetobutylicum, Clostridium beijerinckii, and Clostridium NCP262. PMID- 10812181 TI - Production of antifungal recombinant peptides in Escherichia coli. AB - Antifungal peptides derived from the human bactericidal/permeability-increasing protein (BPI) were produced in Escherichia coli as fusion proteins with human BoneD. Bacterial cultures transformed with the gene encoding the fusion protein were grown to a high cell density (OD(600)>100), and induced with L-arabinose to initiate product expression. Fusion protein accumulated into cytoplasmic inclusion bodies and recombinant peptide was released from BoneD by acid hydrolysis at an engineered aspartyl-prolyl dipeptide linker. Acid hydrolysis of purified inclusion bodies at pH <2.6 followed Arrhenius kinetics and did not require prior inclusion body solubilization in detergents or denaturants. Surprisingly, at pH <2.6 and 85 degrees C, cell lysis and aspartyl-prolyl hydrolysis with concomitant peptide release occurred simultaneously. Bacterial cultures were, therefore, adjusted to approximately pH 2.6 with HCl directly in the bioreactor and incubated at elevated temperature. Peptide, which is soluble in the aqueous acidic environment, was separated from the insoluble material and purified using column separation techniques. Recombinant peptide was separated from the hydrolyzed bioreactor culture with >76% recovery and a final peptide purity of >97%. Antifungal peptide prepared by recombinant and solid phase synthesis methods demonstrated similar activity against Candida sp. in a broth microdilution assay. PMID- 10812182 TI - Determination of metabolic rate-limitations by precursor feeding in Catharanthus roseus hairy root cultures. AB - Precursors from the terpenoid and tryptophan branches were fed to Catharanthus roseus to determine which of the two branches limits metabolic flux to indole alkaloids. The feeding of tryptophan at 17 days of the culture cycle produced auxin-like effects. Addition of low levels of auxin or tryptophan resulted in significant increases in flux to the indole alkaloids. Conversely, feeding higher levels of auxin or tryptophan resulted in increased branching and thickening of the hairy root cultures. A dramatic reduction in flux to the alkaloids was also observed. However, feeding tryptamine or terpenoid precursors had no effect. Therefore, neither pathway tested revealed to be rate-limiting during the late growth phase. Feeding of either geraniol, 10-hydroxygeraniol, or loganin at 21 days each resulted in significant increases in the accumulation of tabersonine. The addition of tryptophan or tryptamine had no effect during the stationary phase of the growth cycle. Thus, during the early stationary phase of growth the terpenoid pathway appears to be rate-limiting. Combined elicitation with jasmonic acid and feeding either loganin or tryptamine did not further enhance the accumulation of indole alkaloids. PMID- 10812183 TI - Bcl-2 mediated suppression of apoptosis in myeloma NS0 cultures. AB - The influence of Bcl-2 expression on the suppression of apoptosis during the cultivation of an NS0 cell line expressing a chimeric antibody was investigated. Following selection of transfectants in medium containing G418, Western analysis revealed evidence of some up-regulation of endogenous Bcl-2 expression even in the control vector transfectants. Cultivation of the two cell lines in suspension batch cultures clearly demonstrated the enhanced robustness of the bcl-2 vector transfected cells. Suppression of apoptosis resulted in an approximately 20% increase in maximum viable cell number, and a doubling in culture duration compared to the control transfected cells. However, despite the significant affect on viability, Bcl-2 expression did not result in an increase in final antibody titre in comparison with the control cell line. Exposure of cells to various nutrient limited conditions further emphasised the influence of Bcl-2 on cell survival. After 3 days of exposure to serum, glucose, glutamate and asparagine deprivation, the viable cell number and viability were significantly higher in the bcl-2 transfected cell line. When control cells were deprived of all amino acids, there was a complete loss of viability and viable cell number within 3 days. By contrast, the bcl-2 transfected cell line retained greater than 75% of the initial viable cell number and about 70% viability. In response to exposure to 8 mM thymidine (a cytostatic agent) the control cell line underwent complete loss of viability and viable cell number after 6 days. This compared with 18 days for complete loss of viability in the bcl-2 transfected cell line. As under batch culture conditions, there was no difference between the two cell lines in final antibody titre, which indicated that MAb synthesis is limited by nutrient availability during the latter stages of culture in both cases. When fed batch cultures were carried out using a concentrated essential amino acid feed, the bcl-2 cell line exhibited a 60% increase in maximum viable cell number and a 50% increase in culture duration, when compared to the control cell line. Moreover, the bcl-2 cell line exhibited a greater than 40% increase in maximum antibody titre. PMID- 10812184 TI - Genetic engineering of the Fusarium solani pisi lipase cutinase for enhanced partitioning in PEG-phosphate aqueous two-phase systems. AB - The Fusarium solani pisi lipase cutinase has been genetically engineered to investigate the influence of C-terminal peptide extensions on the partitioning of the enzyme in PEG-salt based aqueous two-phase bioseparation systems. Seven different cutinase lipase variants were constructed containing various C-terminal peptide extensions including tryptophan rich peptide tags ((WP)(2) and (WP)(4)), positively ((RP)(4)) and negatively ((DP)(4)) charged tags as well as combined tags with tryptophan together with either positively ((WPR)(4)) or negatively ((WPD)(4)) charged amino acids. The modified cutinase variants were stably produced in Escherichia coli as secreted to the periplasm from which they were efficiently purified by IgG-affinity chromatography employing an introduced N terminal IgG-binding ZZ affinity fusion partner present in all variants. Partitioning experiments performed in a PEG 4000/sodium phosphate aqueous two phase system showed that for variants containing either (WP)(2) or (WP)(4) peptide extensions, 10- to 70-fold increases in the partitioning to the PEG rich top-phase were obtained, when compared to the wild type enzyme. An increased partitioning was also seen for cutinase variants tagged with both tryptophans and charged amino acids, whereas the effect of solely charged peptide extensions was relatively small. In addition, when performing partitioning experiments from cell disintegrates, the (WP)(4)-tagged cutinase showed a similarly high PEG-phase partitioning, indicating that the effect from the peptide tag was unaffected by the background of the host proteins. Taken together, the results show that the partitioning of the recombinantly produced cutinase model enzyme could be significantly improved by relatively minor genetic engineering and that the effects observed for purified proteins are retained also in an authentic whole cell disintegrate system. The results presented should be of general interest also for the improvement of the partitioning properties of other industrially interesting proteins including bulk enzymes. PMID- 10812185 TI - Chemoenzymatical grafting of acrylamide onto lignin. AB - Laccase (E.C.1.10.3.2.) from white-rot basidiomycete Trametes versicolor and dioxane peroxides were essential in the copolymerization of acrylamide and a derivative with lignin in a dioxane-H(2)O (7:3) mixture. Both a solubility test and an elemental analysis of the eluted and separated fractions provided evidence of grafting. Gel permeation chromatography on Sephadex G-100 showed that the side chain and the lignin backbone migrate as one unit through the column. Copolymers obtained were characterized by UV-VIS-spectroscopy as well as FT-IR, and 13C CPMAS-NMR spectroscopy. Freeze-dried copolymers of lignin and acrylamide appeared as homogeneous fibril-like particulate. The mechanism of the enzymatical grafting is discussed. PMID- 10812186 TI - Glucagon rDNA origin (GlucaGen) and recombinant LH. (no. 12 in a series of articles to promote a better understanding of the use of genetic engineering). PMID- 10812187 TI - Predictive and prognostic factors in small cell lung cancer: current status. AB - Clinical and laboratory parameters can predict response to chemotherapy and long term survival in small cell lung cancer, and may predict those at risk of early treatment related toxicity. This paper reviews the predictive models that have been developed to divide patients into prognostic groups for response and survival on the basis of clinical and laboratory parameters. These factors may be used for the stratification of patients in clinical trials and to help clinicians make appropriate treatment decisions for individual patients. A number of treatment-related factors can also affect outcomes. The evidence for interventions to prevent treatment deaths in high risk patients, such as prophylactic antibiotics, dosage modification or colony stimulating factor support are also reviewed. PMID- 10812188 TI - Lung cancer and positron emission tomography with fluorodeoxyglucose. AB - Over the past years, positron emission tomography (PET) with fluoro-2-deoxy-D glucose (FDG) has emerged as an important imaging modality. In the thorax, FDG PET has been shown to differentiate benign from malignant pulmonary lesions and stage lung cancer. Preliminary studies have shown its usefulness in assessing tumor recurrence, and assisting in radiotherapy planning. FDG-PET is often more accurate than conventional imaging studies, and has been proven to be cost effective in evaluating lung cancer patients. This review will discuss the current applications of FDG-PET as compared with conventional imaging in diagnosing, staging, and following patients with lung cancer. PMID- 10812189 TI - Computed tomographic diagnosis of bronchogenic carcinoma in HIV-infected patients. AB - PURPOSE: To describe the features of bronchogenic carcinoma (BC) on plain radiography and computed tomography (CT) in human immunodeficiency virus (HIV) infected patients; to evaluate percutaneous transthoracic needle biopsy (PTNB) in this setting; and to assess outcome. PATIENTS AND METHODS: We reviewed the medical charts, radiographs and chest CT scans in 15 AIDS patients with histologically proven BC. All but one of the patients were young men (mean age 48 years) with a long history of smoking (mean 40 pack-years). Adenocarcinoma was the predominant cell type (46.6%). The stage of the malignancy did not correlate with the CD4 cell count (mean 189 per mm(3)). The diagnosis was obtained by means of PTNB (n=7), bronchoscopy (n=4), thoracotomy (n=2), pleural biopsy (n=1) or extrathoracic biopsy (n=1). RESULTS: Parenchymal masses and nodules were the most common features (66.6%) on chest radiographs and CT. BC was peripheral in 11 cases (73%) and was located in the upper lobe in ten cases (66.6%). Enlarged lymph nodes were present in 60% of patients and metastases in 30%. PTNB was diagnostic in seven of the eight patients who underwent the procedure; complications included two pneumothoraces and one secondary implantation of tumor cells along the needle tract. Three lobectomies and one pneumonectomy were performed for stage I disease. The mean survival time among the patients who underwent surgery was 14 months. These survivals are more encouraging than some of those previously reported in the literature, furthermore, patients die of competing illnesses. CONCLUSION: BC in HIV-infected patients is similar to that in the general population. Early diagnosis can be achieved by means of PTNB. Surgical resection, when feasible, significantly improves survival. PMID- 10812191 TI - Aneuploidy of chromosome 7 can be detected in invasive lung cancer and associated premalignant lesions of the lung by fluorescence in situ hybridisation. AB - In the present study the chromosomal status of seven invasive non small cell lung cancer specimens and associated premalignant lesions was investigated. By fluorescence in situ hybridisation (FISH) with centromere specific probes, an increase in the percentage of aneuploid cells from pre-invasive to invasive lesions could be demonstrated (mean 8.5 and 59%, respectively, for chromosome 7). Furthermore, mean chromosome copy numbers were higher in invasive carcinomas as compared to premalignant lesions, indicating polyploidization during tumor development. Increasing evidence suggests that aberrations of chromosome 7 occur early in the development of lung cancer. Whether these aberrations can be used as a biomarker for future neoplastic progression remains to be determined. PMID- 10812190 TI - Allelic imbalance at the 5q14 locus is associated with decreased apoptotic rate in non-small cell lung carcinomas (NSCLCs). Possible synergistic effect with p53 gene alterations on apoptosis. AB - Deletions in the 5q14 region have been described in a variety of neoplasms, such as testicular germ cell tumors, ovarian, gastric and lung cancer. The high frequency of allelic losses observed in this region implies the presence of putative tumor suppressor gene(s) (TSGs). In the present study, we investigated in a series of 56 non-small cell lung carcinomas (NSCLCs) the allelic imbalance (Alm) within the 5q14 region, employing the D5S644 marker, and its relationship with p53 abnormalities, the kinetic parameters [proliferation index (PI) and apoptotic index (AI)] and the ploidy status of the carcinomas. AI at D5S644 was found at a frequency of 51.2%. The rather high percentage of Alm in stage I tumors suggests an early involvement in NSCLC development. LOH at 5q14 was associated with decreased AR in lung tumors insinuating the presence of a putative TSG(s) (P=0.008). Simultaneous alterations of both p53 and D5S644 locus were the most frequent pattern observed (37.5%). Cases demonstrating this profile also exhibited a marked decrease in AI (P=0.001). These findings imply a synergistic mechanism of co-operation between different TSGs. However, proliferation activity was dependent only on p53 status, leading to the assumption that the putative TSG(s) present at 5q14 may probably be involved in normal apoptotic procedures. Further studies are needed to identify the candidate gene(s). PMID- 10812193 TI - Glutamate-glutamine homeostasis in the CNS: physiological and pathophysiological aspects PMID- 10812192 TI - Amifostine plus cisplatin plus vinorelbine in the treatment of advanced non small cell lung cancer: a multicenter phase II study. AB - PURPOSE: to evaluate the activity and toxicity of the combination cisplatin plus vinorelbine plus amifostine in advanced non small cell lung cancer (NSCLC). PATIENTS AND METHODS: a two-stage Simon design was applied. To proceed after the first stage, responses from seven of 19 patients were needed. Overall, 17 responses from 40 treated patients were required to comply with the design parameter. Inclusion criteria were cyto-histologically proven stage IIIB-IV NSCLC; age of 70 years or less; Eastern Cooperative Oncology Group (ECOG) performance status of 2 or less; normal cardiac, hepatic, renal and bone marrow functions; and no previous chemotherapy. Patients were staged by physical examination, biochemistry, chest radiograph, brain, thoracic and abdominal computed tomographic (CT) scans, and bone scan. All patients received cisplatin 100 mg/m(2) intravenously (iv) day 1, vinorelbine 25 mg/m(2) iv days 1-8-15-22, amifostine 740 mg/m(2) iv day 1 every 4 weeks up to six cycles. Eleven of 40 enrolled patients were stage IIIB and 29 stage IV, with a median age of 57 years (range, 38-70 years). RESULTS: all patients were evaluable for response and toxicity (intention to treat analysis). We observed 20 (50%) objective responses, with four (10%) complete responses. Median time to progression was 20 weeks, and median survival was 45 weeks. The toxicity was manageable. The reported main toxicities were neutropenia grade 4 in 10% of patients, grade 1 and grade 3 nephrotoxicity both in 5% of patients and grade 1 amifostine-related hypotension in 15% of patients. CONCLUSION: these data show that cisplatin plus vinorelbine plus amifostine is an active and feaseable regimen in stage IIIB-IV NSCLC. A phase III trial comparing cisplatin plus vinorelbine versus cisplatin plus vinorelbine plus amifostine in advanced NSCLC is warranted. PMID- 10812194 TI - Neuronal-astrocytic and cytosolic-mitochondrial metabolite trafficking during brain activation, hyperammonemia and energy deprivation. AB - A novel concept is described, according to which both neurons and astrocytes are capable of metabolizing glucose all the way to CO(2) and water, but in addition interact metabolically in a process generating glutamate from glucose, and subsequently, metabolizing excess glutamate to CO(2) and water Hertz, L., Dringen, R., Schousboe, A., Robinson, S.R., 1999. Astrocytes: Glutamate producers for neurons (Journal of Neuroscience Research 57, 417-428). The proposed metabolic degradation of glucose via glutamate serves the purpose of adjusting transmitter pools of glutamate to the demands for glutamatergic transmission, and it must account for a major fraction of glucose utilization. Evidence in favor of this concept is presented and a multitude of in vivo data are interpreted in the context of metabolic trafficking between neurons and astrocytes. In addition, intracellular trafficking occurs between cytosol and mitochondria during synthesis of transmitter glutamate, partly explaining a robust quantitative correlation between glutamine synthesis, as a measure of release of transmitter glutamate, and glucose utilization, reported by several authors. Both intracellular and intercellular metabolic trafficking may be affected during pathological conditions, as evidenced by effects of hyperammonemia (mimicking hepatic encephalopathy) and energy deprivation (mimicking stroke). It is suggested that neuronal-astrocytic interactions may also be impaired during degenerative dementing diseases. PMID- 10812195 TI - Mechanisms of action of valproate: a commentatory. AB - Valproate, one of the major antiepileptic drugs used today, has besides its wide use in both generalized and partial epilepsies, several new approved indications including the treatment of bipolar disorders, neuropathic pain, and as a migraine prophylaxis. This wide spectrum of activities is reflected by several different mechanisms of action, which are discussed in this review. With regard to the antiepileptic effect of VPA, a special emphasis is put on the effect on the GABAergic system and the effect on enzymes like succinate semialdehyde dehydrogenase (SSA-DH), GABA transaminase (GABA-T), and alpha-ketoglutarate dehydrogenase, related to the tricarboxylic acid (TCA) cycle and thereby cerebral metabolism. In vitro studies have shown that VPA is a potent inhibitor of SSA-DH. In brain homogenates, GABA-T is inhibited at high concentrations only. Besides affecting the GABA-shunt, VPA might also inhibit the TCA cycle at the alpha ketoglutarate dehydrogenase step. The effect of VPA on excitatory neurotransmission and on excitatory membranes are mechanisms likely to be responsible for the 'mood-stabilizing' effect as well as in the treatment of migraine. GABA-mediated responses may be involved in neuropathic pain. But still there are many aspects of the mechanisms of action of VPA that remain unknown. PMID- 10812196 TI - Structural features and regulatory properties of the brain glutamate decarboxylases. AB - It is widely recognized that the two major forms of GAD present in adult vertebrate brains are each composed of two major sequence domains that differ in size and degree of similarity. The amino-terminal domain is smaller and shows little sequence identity between the two forms. This domain is thought to mediate the subcellular targeting of the two GADs. Substantial parts of the amino terminal domain appear to be exposed and flexible, as shown by proteolysis experiments and the locations of posttranslational modifications. The carboxyl terminal sequence domain contains the catalytic site and shows substantial sequence similarity between the forms. The interaction of GAD with its cofactor, pyridoxal-5' phosphate (pyridoxal-P), plays a key role in the regulation of GAD activity. Although GAD(65) and GAD(67) interact differently with pyridoxal-P, their cofactor-binding sites contain the same set of nine putative cofactor binding residues and have the same basic structural fold. Thus the cofactor binding differences cannot be attributed to fundamental structural differences between the GADs but must result from subtle modifications of the basic cofactor binding fold. The presence of another conserved motif suggests that the carboxyl terminal domain is composed of two functional domains: the cofactor-binding domain and a small domain that closes when the substrate binds. Finally, GAD is a dimeric enzyme and conserved features of GADs superfamily of pyridoxal-P proteins indicate the dimer-forming interactions are mediated mainly by the carboxyl terminal domain. PMID- 10812197 TI - Functional integration of the transport of ammonium, glutamate and glutamine in astrocytes. AB - This short review surveys the effects of extracellular potassium, released by neuronal activity, on the fluxes of ammonium, glutamate and glutamine in astrocytes. There is evidence that each of these fluxes is modulated by potassium induced changes in astrocytic pH. The result is viewed as an integrated response to neuronal activity. The unusually high permeability of astrocyte cell membrane to ammonium ions, together with the normal transmembrane gradient of pH, enables astrocytes to accumulate ammonium appreciably. However, at loci of neuronal activity, effective ammonium ion permeability is diminished and the cytosol is alkalinized, resulting in a local decline in intracellular ammonium concentration. Intracellular potassium concentration rises at these same loci, creating the conditions for a 'potassium-ammonium countercurrent' in which ammonium ions migrate intracellularly towards sites of neuronal activity as potassium ions diffuse away. Physiologic elevations of extracellular potassium evoke a marked 'paradoxical' increase in the velocity of glutamate uptake in astrocytes. This increase correlates well with the extent of potassium-induced alkalinization. Further, recent evidence identifies a major transporter of glutamine in astrocytes (System N) as a glutamine/proton exchanger. Potassium can reverse the transmembrane gradient of protons in astrocytes, and increase intracellular glutamine concentration, creating the conditions for a reversal of glutamine flux via System N from uptake to export. These flux changes, evoked by potassium released from active neurons, combine to accelerate glutamate-glutamine cycling. PMID- 10812198 TI - Glutamine transport in brain mitochondria. AB - Gln is transported into rat brain synaptic and non-synaptic mitochondria by a protein catalyzed process. The uptake is significantly higher in synaptic than in non-synaptic mitochondria. The transport is inhibited by the amino acids Glu, Asn and Asp, and by the TCA cycle intermediates succinate, malate and 2-OG. The inhibition by 2-OG is counteracted by AOA and is therefore assumed to be due to transamination of 2-OG, whereby Glu is formed. This presumes that Glu also binds to an inhibitory site on the matrix face of the inner membrane. The transport is complex and cannot be explained by the simple uniport mechanism which has been proposed for renal (Schoolwerth and LaNoue, 1985), and liver mitochondria (Soboll et al., 1991). Thus, Gln transport is stimulated by respiration and by the proton electrochemical gradient. Since it is indicated that both the neutral Gln zwitterion and the Gln anion are transported, there are probably different uptake mechanisms, but not necessarily different carriers. Gln may be transported by an electroneutral mechanism as a proton compensated anion, as well as electrophoretically as a zwitterion with a proton, and probably also by diffusion as a zwitterion. The properties of the brain mitochondrial Gln uptake mechanisms are also not identical with those of a purified renal Gln transporter. It is possible that the Gln transport is controlled by more than one protein, which may be situated on distinct species in a heterogeneous mitochondrial population. Since Gln is assumed to participate in energy production as well as in the synthesis of nucleic acid components and proteins in brain mitochondria, the control of Gln uptake in these organelles may be important. PMID- 10812199 TI - Glutamine transport in C6 glioma cells. AB - Glutamine transport across the cell membranes of a variety of mammalian tissues is mediated by at least four transport systems: a sodium-independent system L, and sodium-dependent systems A, ASC and N, the latter occurring in different tissue-specific variants. In this study we assessed the contribution of these systems to the uptake of [(3)H]glutamine in C6 rat glioma cells. The sodium dependent uptake, which accounted for more than 80% of the total uptake, was not inhibited by 2-methylaminoisobutyric acid (MeAIB), indicating that system A was inactive, possibly being depressed by glutamine present in the culture medium. About 80% of the sodium-dependent uptake was mediated by system ASC, which differed from system ASC common to other CNS- and non-CNS tissues by its pH dependence and partial lithium tolerance. The residual 20% of sodium-dependent uptake appeared to be mediated by system N, which was identified as a component resistant to inhibition by MeAIB+threonine. The system N in C6 cells appeared to be neither fully compatible with the neuronal system Nb, nor with the N system described in astrocytes: it differed from the former in being strongly inhibited by histidine and showing fair tolerance for lithium, and from the latter in its pH-insensitivity and strong inhibition by glutamate. The sodium-independent glutamine uptake differed from the astrocytic or neuronal uptake in its relatively weak inhibition by system L substrates and a strong inhibition by system ASC substrates, indicating a possible contribution of a variant of the ASC system. PMID- 10812200 TI - Substrate-induced up-regulation of Na(+)-dependent glutamate transport activity. AB - Sodium-dependent transporters regulate extracellular glutamate in the CNS. Recent studies suggest that the activity of several different neurotransmitter transporters can be rapidly regulated by a variety of mechanisms. In the present study, we report that pre-incubation of primary 'astrocyte-poor' neuronal cultures with glutamate (100 microM) for 30 min nearly doubled the V(max) for Na(+)-dependent accumulation of L-[(3)H]-glutamate, but had no effect on Na(+) dependent [(3)H]-glycine transport. Pre-incubation with glutamate also increased the net uptake of non-radioactive glutamate, providing evidence that the increase in accumulation of L-[(3)H]-glutamate was not related to an increase in intracellular glutamate and a subsequent increase in exchange of intracellular non-radioactive glutamate for extracellular radioactive glutamate. The glutamate receptor agonists, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate, quisqualate, and (1 S, 3R)-1-aminocyclopentane-1,3-dicarboxylic acid did not mimic the effect of pre-incubation with glutamate and the glutamate-induced increase was not blocked by receptor antagonists. However, compounds known to interact with the transporters, including L-aspartate, D-aspartate, L-(-)-threo-3 hydroxyaspartate (L-THA) and L-trans-pyrrolidine-2,4-dicarboxylate (L-trans-PDC), caused variable increases in transport activity and attenuated the increase induced by glutamate, suggesting that the increase is related to the interaction of glutamate with the transporters. Several studies were attempted to define the mechanism of this regulation. We found no evidence for increases in transporter synthesis or cell surface expression. Inhibitors of signaling molecules known to regulate other neurotransmitter transporters had no effect on this stimulation. Using a variety of cultures, evidence is provided to suggest that this substrate induced up-regulation of glutamate transport is specific for the GLT-1 and GLAST subtypes and does not influence transport mediated by EAAC1. These studies suggest that the interaction of glutamate with some of the subtypes of glutamate transporters causes an increase in transport activity. Conceivably, this phenomenon provides an endogenous mechanism to increase the clearance of glutamate during periods of prolonged elevations in extracellular glutamate. PMID- 10812201 TI - The high-affinity glutamate transporters GLT1, GLAST, and EAAT4 are regulated via different signalling mechanisms. AB - High-affinity glutamate transporters ensure termination of glutamatergic neurotransmission and keep the synaptic concentration of this amino acid below excitotoxic levels. However, neuronal glutamate transporters, EAAC1 and EAAT4, are located outside the synaptic cleft and contribute less significantly to the glutamate uptake in the brain than two astroglial transporters, GLAST and GLT1. Aberrant functioning of the glutamate uptake system seems to be linked to some neurodegenerative disorders (eg amyotrophic lateral sclerosis, ALS). Expression of glutamate transporters is differentially regulated via distinct cellular mechanisms. GLT1, which is expressed at very low levels in cultured astrocytes, is strongly induced in the presence of neurons. The present immunocytochemical data provide further evidence that neuronal soluble factors, rather than physical contact between neurons and glia, determine the induction of GLT1 in astrocytes. This effect is apparently mediated by yet undefined growth factor(s) via the tyrphostin-sensitive receptor tyrosine kinase (RTK) signalling, that in turn, supports the downstream activation of p42/44 MAP kinases and the CREM and ATF-1 transcription factors. RTK-independent simultaneous activation of the CREB transcription factor suggests a possible involvement of complementary pathway(s). Neuronal soluble factors do not affect expression of GLAST, but induce supporting machinery for differential regulation of GLAST via the astroglial metabotropic glutamate receptors, mGluR3 and mGluR5. Thus, long-term treatment with the group I mGluR agonist, DHPG, causes down-regulation of GLAST, whereas the group II agonist, DCG-IV, has an opposite effect on the expression of GLAST in astrocytes. However, in BT4C glioma cells glutamate or other transportable substrates (D aspartate and L-2,4-trans-PDC) induced cell-surface expression of EAAT4 in a receptor-independent manner. The activity-dependent trafficking of this transporter which also exhibits properties of a glutamate-gated chloride channel may play functional roles not only in neuronal excitability, but in glioma cell biology as well. PMID- 10812202 TI - Persistent depolarization and Glu uptake inhibition operate distinct osmoregulatory mechanisms in the mammalian brain. AB - The ways of coupling neuronal with glial compartments in natural physiology was investigated in microdialysis experiments by monitoring extracellular concentration of amino acids in the brain of anaesthetized rats. We hypothesized that extracellular [Glu], [Gln] and [Tau] patterns would be state-dependent. This was tested by stimulation of N-methyl-D-aspartate (NMDA) receptors, by inhibition of Glu uptake or by local depolarization with a high-K(+) dialysate, coupled with the addition of Co(2+) to block Ca(2+) influx. The results showed that (1) extracellular [Gln] was low whereas [Glu] and [Tau] were high during infusion of NMDA (0.5-1.0 mM) or high-K(+) (80 mM) in the hippocampus and ventrobasal thalamus, (2) hippocampal extracellular [Glu], [Gln] and [Tau] were increased in response to the Glu uptake inhibitor, L-trans-pyrrolidine-2, 4-dicarboxilic acid (tPDC, 0.5-3.0 mM), in a concentration-dependent manner, (3) high-K(+)-induced increase of extracellular [Glu] was partially blocked by the addition of 10 mM CoCl(2) with the high-K(+) dialysate in the hippocampus. Searching for main correlations between changes in [Glu], [Gln] and [Tau] by calculating partial correlations and with the use of factor analyses we found, the primary response of the mammalian brain to persistent depolarization is the neuronal uptake of [Gln] and release of [Tau] thereupon, acting independently of Glu changes. When glial and neuronal uptake of Glu is blocked, releases of Tau occur from neuronal as well as glial compartments accompanied by increases of [Gln] in the mammalian brain. PMID- 10812203 TI - Fine tuning of glutamate uptake and degradation in glial cells: common transcriptional regulation of GLAST1 and GS. AB - Glutamate is the major excitatory neurotransmitter in the mammalian brain and retina, and glutamate uptake is essential for the normal function of glutamatergic synapses in the retina. As summarized here, all neuronal and macroglial cells of the retina express high-affinity glutamate transporters. GLAST1 is expressed in glial cells, whereas GLT1 and EAAC1 are neuronal. Glutamate uptake studies in intact retina revealed that Muller glial cells dominate the total retinal glutamate transport and that this uptake is strongly influenced by the activity of glutamine synthetase. A prerequisite for an effective glutamate-glutamine cycle in glial cells would be the regulated coordination between glutamate uptake and glutamate degradation. Using cultured retinal Muller glial cells, we demonstrate that protein expression of both, GLAST1 and glutamine synthetase, are inducible by the glucocorticoid hormone cortisol. These results suggest a common transcriptional regulation of the key proteins in the glial portion of the glutamate-glutamine cycle and may impact on transmitter clearance, transmitter recycling and, as discussed, on the development of the cellular architecture in retina. PMID- 10812204 TI - Are neuronal transporters relevant in retinal glutamate homeostasis? AB - Exposure of isolated retinas to 30 microM D-aspartate, which is a substrate for all high affinity glutamate transporters, for 30 min, resulted in the accumulation of such D-aspartate into Muller glial cells but not glutamatergic neurons as evinced by immunocytochemistry for D-aspartate. Further incubation of such loaded retinas in physiological media, in the absence of D-aspartate, resulted in the slow release of accumulated D-aspartate from the Muller cells and its accumulation into populations of photoreceptors and bipolar cells. This result indicates that after initial transport into Muller cells, reversal of direction of transport of D-aspartate, and thus by inference glutamate, by GLAST, readily occurs. D-aspartate released by Muller cells was strongly accumulated into cone photoreceptors which are known to express GLT-1, and into rod photoreceptors which we demonstrate here to express the retina specific glutamate transporter EAAT5 (excitatory amino transporter 5). Populations of glutamatergic bipolar cells, which express GLT-1 also exhibited avid uptake of D-aspartate. We conclude that the Muller cell glutamate transporter GLAST is responsible for most of the initial glutamate clearance in the retina after its release from neurones. However, some glutamate is also returned from Muller cells, to neurons expressing GLT-1 and EAAT5, albeit at a slow rate. These data suggest that the role of neuronal glutamate transporters in the retina may be to facilitate a slow process of recycling glutamate back from Muller cells to neurons after its initial clearance from perisynaptic regions by GLAST. PMID- 10812205 TI - Methylmercury alters glutamate transport in astrocytes. AB - Methylmercury (MeHg) is a significant environmental contaminant that will continue to pose great risk to human health. Considerable attention in the scientific and health policy fora is focused on the question of whether MeHg intake from a diet high in fish is associated with aberrant CNS function. A number of recent studies (Kjellstrom et al., 1986: Kjellstrom, T., Kennedy, P., Wallis, S., Mantell, C., 1986. Physical and mental development of children with prenatal exposure to mercury from fish. Stage I: preliminary tests at age 4. Solna, Sweden. National Swedish Environmental Protection Board Report 3080, 1989: Kjellstrom, T., Kennedy, P., Wallis, S., Stewart, A., Friberg, L. et al., 1989. Physical and mental development of children with prenatal exposure to mercury from fish. Stage II: interviews and psychological tests at age 6. Solna, Sweden. National Swedish Environmental Protection Board Report 3642; McKeown-Eyssen et al., 1983: McKeown-Eyssen, G., Ruedy, J., Neims, A. , 1983. Methylmercury exposure in Northern Quebec II: neurologic findings in children. American Journal of Epidemiology 118, 470-479; Grandjean et al., 1997: Grandjean, P., Weihe, P., White, R. F., Debes, F., Araki, S., Yokoyama, K., Murata, K., Sorensen, N., Dahl, R., Jorgensen, P. J., 1997. Cognitive deficit in 7-year-old children with prenatal exposure to methylmercury. Neurotoxicology and Teratology 19, 417-428) suggest that fetal exposure at levels attained by mothers eating fish regularly during pregnancy are associated with neurological deficits in their offspring. Astrocytes play a key role in MeHg-induced excitotoxicity. (1) MeHg preferentially accumulates in astrocytes. (2) MeHg potently and specifically inhibits glutamate uptake in astrocytes. (3) Neuronal dysfunction is secondary to disturbances in astrocytes. (4) Co-application of nontoxic concentrations of MeHg and glutamate leads to the typical appearance of neuronal lesions associated with excitotoxic stimulation. (5) MeHg induces swelling of astrocytes. These observations are fully consistent with MeHg-induced dysregulation of excitatory amino acid homeostasis, and indicate that a glutamate-mediated excitotoxic mechanism is involved. This manuscript details the role of astrocytes in mediating MeHg-induced excitotoxicity, and elaborates on the protective role afforded by metallothioneins (MTs) in attenuating MeHg cytotoxicity. PMID- 10812206 TI - Effects of thiopental on transport and metabolism of glutamate in cultured cerebellar granule neurons. AB - This study was performed to analyze the effects of the barbiturate thiopental on neuronal glutamate uptake, release and metabolism. Since barbiturates are known to bind to the GABA(A) receptor, some experiments were carried out in the presence of GABA. Cerebellar granule neurons were incubated for 2 h in medium containing 0.25 mM [U-(13)C]glutamate, 3 mM glucose, 50 microM GABA and 0.1 or 1 mM thiopental when indicated. When analyzing cell extracts, it was surprisingly found that in addition to glutamate, aspartate and glutathione, GABA was also labeled. In the medium, label was observed in glutamate, aspartate and lactate. Glutamate exhibited different labeling patterns, indicating metabolism in the tricarboxylic acid cycle, and subsequent release. A net uptake of [U (13)C]glutamate and unlabeled glucose was seen under all conditions. The amounts of most metabolites synthesized from [U-(13)C]glutamate were unchanged in the presence of GABA with or without 0.1 mM thiopental. In the presence of 1 mM thiopental, regardless of the presence of GABA, decreased amounts of [1,2, 3 (13)C]glutamate and [U-(13)C]aspartate were found in the medium. In the cell extracts increased [U-(13)C]glutamate, [1,2, 3-(13)C]glutamate, labeled glutathione and [U-(13)C]aspartate were observed in the 1 mM thiopental groups. Glutamate efflux and uptake were studied using [(3)H]D-aspartate. While efflux was substantially reduced in the presence of 1 mM thiopental, this barbiturate only marginally inhibited uptake even at 3 mM. These results may suggest that the previously demonstrated neuroprotective action of thiopental could be related to its ability to reduce excessive glutamate outflow. Additionally, thiopental decreased the oxidative metabolism of [U-(13)C]glutamate but at the same time increased the detectable metabolites derived from the TCA cycle. These latter effects were also exerted by GABA. PMID- 10812207 TI - Metabolism of (1-(13)C) glucose and (2-(13)C, 2-(2)H(3)) acetate in the neuronal and glial compartments of the adult rat brain as detected by [(13)C, (2)H] NMR spectroscopy. AB - Ex vivo ?(13)C, (2)H? NMR spectroscopy allowed to estimate the relative sizes of neuronal and glial glutamate pools and the relative contributions of (1-(13)C) glucose and (2-(13)C, 2-(2)H(3)) acetate to the neuronal and glial tricarboxylic acid cycles of the adult rat brain. Rats were infused during 60 min in the right jugular vein with solutions containing (2-(13)C, 2-(2)H(3)) acetate and (1-(13)C) glucose or (2-(13)C, 2-(2)H(3)) acetate only. At the end of the infusion the brains were frozen in situ and perchloric acid extracts were prepared and analyzed by high resolution (13)C NMR spectroscopy (90.5 MHz). The relative sizes of the neuronal and glial glutamate pools and the contributions of acetyl-CoA molecules derived from (2-(13)C, (2)H(3)) acetate or (1-(13)C) glucose entering the tricarboxylic acid cycles of both compartments, could be determined by the analysis of (2)H-(13)C multiplets and (2)H induced isotopic shifts observed in the C4 carbon resonances of glutamate and glutamine. During the infusions with (2 (13)C, 2-(2)H(3)) acetate and (1-(13)C) glucose, the glial glutamate pool contributed 9% of total cerebral glutamate being derived from (2-(13)C, 2 (2)H(3)) acetyl-CoA (4%), (2-(13)C) acetyl-CoA (3%) and recycled (2-(13)C, 2 (2)H) acetyl-CoA (2%). The neuronal glutamate pool accounted for 91% of the total cerebral glutamate being mainly originated from (2-(13)C) acetyl-CoA (86%) and (2 (13)C, 2-(2)H) acetyl-CoA (5%). During the infusions of (2-(13)C, 2-(2)H(3)) acetate only, the glial glutamate pool contributed 73% of the cerebral glutamate, being derived from (2-(13)C, 2-(2)H(3)) acetyl-CoA (36%), (2-(13)C, 2-(2)H) acetyl-CoA (27%) and (2-(13)C) acetyl-CoA (10%). The neuronal pool contributed 27% of cerebral glutamate being formed from (2-(13)C) acetyl-CoA (11%) and recycled (2-(13)C, 2-(2)H) acetyl-CoA (16%). These results illustrate the potential of ?(13)C, (2)H? NMR spectroscopy as a novel approach to investigate substrate selection and metabolic compartmentation in the adult mammalian brain. PMID- 10812208 TI - Differential distribution of the enzymes glutamate dehydrogenase and aspartate aminotransferase in cortical synaptic mitochondria contributes to metabolic compartmentation in cortical synaptic terminals. AB - There have been numerous studies on the activity and localization of aspartate aminotransferase (AAT) and glutamate dehydrogenase (GDH) in brain tissue. However, there is still a controversy as to the specific roles and relative importance of these enzymes in glutamate and glutamine metabolism in astrocytes and neurons or synaptic terminals. There are many reports documenting GDH activity in synaptic terminals, yet the misconception that it is a glial enzyme persists. Furthermore, there is evidence that this tightly regulated enzyme may have an increased role in synaptic metabolism in adverse conditions such as low glucose and hyperammonemia that could compromise synaptic function. In the present study, we report high activity of both AAT and GDH in mitochondrial subfractions from cortical synaptic terminals. The relative amount of GDH/AAT activity was higher in SM2 mitochondria, compared to SM1 mitochondria. Such a differential distribution of enzymes can contribute significantly to the compartmentation of metabolism. There is evidence that the metabolic capabilities of the SM1 and SM2 subfractions of synaptic mitochondria are compatible with the compartments A and B of neuronal metabolism proposed by Waagepetersen et al. (1998b. Dev. Neurosci. 20, 310-320). PMID- 10812209 TI - Effects of ammonia on glutamate transporter (GLAST) protein and mRNA in cultured rat cortical astrocytes. AB - Ammonia is a neurotoxic substance which accumulates in brain in liver failure and it has been suggested that ammonia plays a key role in contributing to the astrocytic dysfunction characteristic of hepatic encephalopathy. In particular, the effects of ammonia may be responsible for the reduced astrocytic uptake of neuronally-released glutamate and high extracellular glutamate levels consistently seen in experimental models of hepatic encephalopathy. To further address this issue, [(3)H]-D-aspartate uptake was examined in primary rat cortical astrocyte cultures exposed to 5 mM ammonium chloride for a period of 7 days. In addition, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot studies were performed to examine the mRNA and protein expression respectively of the glutamate transporter GLAST in ammonia-treated cells. Studies revealed a 57% (p<0.05) decrease in [(3)H]-D-aspartate uptake and a concomitant significant decrease in GLAST transporter protein (43%, p<0.05) and mRNA (32%, p<0.05) expression. The reduced capacity of astrocytes to reuptake glutamate following ammonia exposure may result in compromised neuron-astrocyte trafficking of glutamate and could thus contribute to the pathogenesis of the cerebral dysfunction characteristic of hyperammonemic syndromes such as hepatic encephalopathy. PMID- 10812210 TI - Differential effects of acute and chronic hyperammonemia on signal transduction pathways associated to NMDA receptors. PMID- 10812212 TI - Characterization of cystine uptake in cultured astrocytes. AB - Glutathione is involved in the maintenance of the structural and functional integrity of membrane proteins, in protection against free radicals and oxidative stress, and in the detoxification of xenobiotics. The cellular uptake of cystine is the rate limiting step in the biosynthesis of glutathione. The precise mechanism for such uptake is not clear as some reports indicate that the uptake occurs through a glutamate-cystine antiporter (system X(c)(-)), whereas, others suggest that it is taken up by the glutamate transporter (system X(AG)). Our studies in cultured astrocytes derived from neonatal rats showed that glutamate, D- and L-aspartate inhibited cystine uptake; that factors that increased intracellular glutamate levels, which would have enhanced the activity of the antiporter, did not stimulate cystine uptake; that the uptake was sodium dependent and partially chloride dependent; that the b(o,+) and ASC systems, which have been shown to carry cystine in some cells, did not mediate cystine uptake in astrocytes; that glutamate uptake blockers such as L-aspartate-beta hydroxamate (AbetaH) and L-trans-pyrrolidine-2,4-dicarboxylate (PDC), as well as cystine uptake inhibitor L-alpha-aminoadipate (AAA) potently reduced cystine uptake. Additionally, deferoxamine (100 microM) as well as ammonium chloride (5 mM), both of which inhibit glutamate uptake, also inhibited cystine uptake. Taken together, our findings indicate that astrocytes take up cystine through a similar, if not identical, system used to take up glutamate. Interference of cystine uptake by astrocytes through the glutamate transport system may have profound effects on the redox state and the structural and functional integrity of the CNS. PMID- 10812211 TI - Elevation of glutathione levels by ammonium ions in primary cultures of rat astrocytes. AB - It is well established that ammonia is detoxified in the brain to form glutamine and that astrocytes play a major role in this process. The synthesis of glutamine requires glutamate and ATP. Since glutamate and ATP are also required for the synthesis of glutathione (GSH), we examined the effect of pathophysiological concentrations of ammonia on levels of GSH in primary cultures of astrocytes. GSH content in the medium increased in a dose- and time-dependent manner in the presence of ammonia. After an initial decrease, cellular GSH content increased in a similar manner. The levels of glutathione disulfide (GSSG) were also increased. A linear relationship was observed between ammonia concentration and the increase in GSH levels. An increase in the efflux of GSH from cells into medium was also observed under these conditions. Buthionine sulfoximine and acivicin, but not methionine sulfoximine, blocked the ammonia induced increase in GSH levels. No, or minor, changes in the activities of enzymes (gamma-glutamyl transpeptidase, GSH reductase and GSH-peroxidase) that might influence GSH levels were identified and thus could not account for the ammonia induced increase in GSH levels in astrocytes. These findings indicate that pathophysiological concentrations of ammonium ions result in increased astroglial levels of GSH which may affect the metabolism and function of astrocytes. PMID- 10812213 TI - Ischemia induced changes in expression of the astrocyte glutamate transporter GLT1 in hippocampus of the rat. AB - Changes in cellular uptake of glutamate following transient cerebral ischemia is of possible importance to ischemia induced cell death. In the present study, we employed in situ hybridization and immunohistochemistry to investigate the influence of cerebral ischemia on expression of mRNA and protein of the astrocyte glutamate transporter GLT1, and of glial fibrillary acidic protein. Different subfields of CA1 and CA3 of the rat hippocampus were studied at various time points after ischemia (days 1, 2, 4, and 21). In CA1, GLT1-mRNA was decreased at all time-points after ischemia except from day 2, whereas in CA3, decreases were seen only on day 1. Expression of GLT1-protein in CA1 was unchanged during the initial days after ischemia, but decreased markedly from day 2 to 4. In CA3, GLT1 protein increased progressively throughout the observation period after ischemia. Following the degeneration of CA1 pyramidal cells, a positive correlation between the number of CA1 pyramidal cells and expression of either GLT1-mRNA or -protein was evident selectively in CA1. Increases in expression of mRNA and protein of glial fibrillary acidic protein were present from day 2, most notable in CA1. The present data provide evidence that expression of GLT1 in CA1 of the hippocampus is not decreased persistently before the degeneration of CA1 pyramidal cells, but is downregulated in response to loss of these neurons. Since the reduction in GLT1 expression evolved concomitantly with the degeneration of CA1 pyramidal cells, it may contribute to the severity of CA1 pyramidal cell loss. A progressive postischemic increase in GLT1 expression in CA3 may be linked to the resistance of CA3 neurons to ischemic cell damage. PMID- 10812214 TI - Delayed induction of apoptosis by ammonia in C6 glioma cells. AB - Ammonia is a neurotoxin whose administration in large doses causes coma and death of the exposed animals, but whether and in what degree these whole body effects are related to the death of CNS cells is not known. Since the downstream effects of ammonia in cultured CNS cells appear to be partly mediated by overactivation of several putative signalling mechanisms characteristic for the apoptotic program, we speculated that ammonia neurotoxicity may be apoptogenic. In this study, C6 glioma cells grown in 2% serum were exposed to 5 mM or 10 mM NH(4)Cl (ammonia) for 96 h and tested for the appearance of apoptosis by (a) Hoechst staining, (b) TUNEL reaction and (c) DNA ladder, at different times of exposure. In cultures exposed to either 5 mM or 10 mM ammonia, about 10% of the cells were found to enter apoptosis at 48 h of exposure, and the number of apoptotic cells rose to 30% at 72 h, and to 50% at 96 h of exposure, respectively. The first transduction signal purportedly involved in apoptosis, activation of PKCalphabeta, was transient and appeared already after 3-6 h of treatment. Coincident with pronounced manifestation of apoptosis (at 72 h and even more at 96 h of exposure) was an increased transfer of the transcription factor NFkappaB from cytoplasmto nucleus as revealed by EMSA assay. The number of cells affected by ammonia-induced apoptosis was markedly reduced by incubation with a NOS inhibitor, L-NAME at 100 microM concentration. The results indicate that ammonia induced apoptosis is a result of a complex interplay of at least three signalling molecules: NO, PKC and the transcription factor NFkappaB, with NFkappaB being possibly involved in the induction of iNOS and generation of toxic levels of NO in the cells. PMID- 10812215 TI - Modulation of glutamate receptor functions by glutathione. AB - In addition to its well-known antioxidant effects, glutathione apparently has an additional double role in the central nervous system as a neurotransmitter and neuromodulator. A number of recent neurochemical, neuropharmacological and electrophysiological studies have yielded evidence on both functions. As an excitatory neurotransmitter, glutathione depolarizes neurons by acting as ionotropic receptors of its own which are different from any other excitatory amino acid receptors. As a neuromodulator, it displaces ionotropic glutamate receptor ligands from their binding sites and regulates calcium influx through N methyl-D-aspartate receptor-governed ionophores. In brain slices glutathione has been shown to regulate the release of other transmitters, e.g., gamma aminobutyrate and dopamine, mediated by N-methyl-D-aspartate receptors. In the present article, we review recent findings on the neuromodulatory actions of glutathione and discuss possible physiological and pathophysiological consequences. PMID- 10812217 TI - Astroglia and glutamate in physiology and pathology: aspects on glutamate transport, glutamate-induced cell swelling and gap-junction communication. AB - Astroglia have the capacity to monitor extracellular glutamate (Glu) and maintain it at low levels, metabolize Glu, or release it back into the extracellular space. Glu can induce an increase in astroglial cell volume with a resulting decrease of the extracellular space, and thereby alter the concentration of extracellular substances. Many lines of evidence show that K(+) can be buffered within the astroglial gap-junction-coupled network, and recent results show that gap junctions are permeable for Glu. All these events occur dynamically: the astroglial network has the capacity to interfere actively with neurotransmission, thereby contributing to a high signal-to-noise ratio for the Glu transmission. High-quality neuronal messages during normal physiology can then be maintained. With the same mechanisms, astroglia might exert a neuroprotective function in situations of moderately increased extracellular Glu concentrations, i.e., corresponding to conditions of pathological hyper-excitability, or corresponding to early stages of an acute brain injury. If the astroglial functions are failing, neuronal dysfunction can be reinforced. PMID- 10812216 TI - NMDA-induced 45Ca release in the dentate gyrus of newborn rats: in vivo microdialysis study. AB - This in vivo study, aimed at detecting the N-methyl-D-aspartate (NMDA) evoked Ca(2+)-induced Ca(2+) release from intracellular stores in the neonatal rat brain, demonstrates that the application of 5 mM N-methyl-D-aspartate via a microdialysis probe for 20 min to the dentate gyrus (DG) of halotane-anesthetized 7 day-old (postnatal day 7, PND 7) rats induces a prolonged decrease in Ca(2+) concentration in an initially calcium-free dialysis medium, indicative of a drop in the extracellular concentration of Ca(2+) and Ca(2+) influx to neurons. In parallel experiments, a huge NMDA-evoked release of 45Ca from the pre-labeled endogenous Ca(2+) pool was observed and interpreted as the expression of intracellular Ca(2+) release. Dantrolene (100 microM) significantly inhibited the NMDA-induced 45Ca release, whereas 250 microM ryanodine exerted an unspecific biphasic effect. Autoradiographic and immunocytochemical detection of ryanodine receptors and calbindin D(28K), respectively, in the hippocampal region of PND 7 rats displayed a pronounced expression of [3H]ryanodine binding sites in the DG, but only a slight immunoreactivity of calbindin D(28K). Plastic changes in neurons or excitotoxic neuronal damage induced by the activation of NMDA receptors are mediated by Ca(2+) signals, resulting from an influx of extracellular Ca(2+), and also in some neurons, from the release of intracellular Ca(2+). Our previous in vivo microdialysis experiments visualized NMDA-evoked 45Ca release in the adult rat dentate gyrus, attributable to Ca(2+)-induced Ca(2+) release from the ryanodine-sensitive pool. An additional role of calbindin in the mechanism of this phenomenon has been suggested. This aspect has not been studied in vivo in newborn rats. Our present results indicate that the release of 45Ca from the prelabeled intracellular, dantrolene-sensitive Ca(2+) pool in the DG neurons of immature rats, most probably representing a phenomenon of Ca(2+) induced Ca(2+) release, significantly participates in the generation of NMDA receptor-mediated intracellular Ca(2+) signals, whereas the role of calbindin D(28K) in the mechanism of 45Ca release is negligible. PMID- 10812218 TI - Analysis of varicella vaccine breakthrough rates: implications for the effectiveness of immunisation programmes. AB - The objective of this study was to quantify key parameters describing varicella zoster virus (VZV) vaccine efficacy. To do so a mathematical model was developed to represent breakthrough cases as a function of time after vaccination in vaccine efficacy trials. Efficacy parameter sets were identified by fitting the predicted annual number of breakthrough infections with that observed in three clinical trials chosen to represent the plausible range of vaccine efficacy. Results suggest that varicella vaccination seems to result in a high proportion of individuals who are initially totally protected (97% for the base-case). However, individuals lose full protection relatively rapidly (3% per year for the base-case). Once total protection has waned individuals have a high probability of developing a breakthrough infection if exposed to varicella (73% of the probability in unvaccinated susceptibles for the base-case). Results also highlight that vaccine efficacy parameters should be estimated concurrently to take into account dependencies between parameters. PMID- 10812219 TI - Protection against influenza virus infection in mice immunized by administration of hemagglutinin-expressing DNAs with electroporation. AB - Electroporation for the transfer of plasmid DNA encoding influenza virus hemagglutinin (HA) into muscle or nasal mucosa was tried in BALB/c mice to examine the efficacy of this method for inducing anti-HA immune responses and providing protection against homologous A/PR/8/34 (PR8) virus infection. Mice were immunized by two injections, 3 weeks apart, of HA-DNA with electroporation into the muscle wherein a pair of electrode needles was inserted to deliver the electric pulses. One or 3 weeks after the immunization, the mice were infected with a lethal dose of the PR8 virus. Ten micrograms or more of HA-DNA/dose induced strong serum anti-HA IgG antibody (Ab) responses, in which both IgG1 and IgG2a were predominant, and weak cytotoxic T lymphocyte responses. These immune responses were sufficient to provide efficient protection against the lethal infection. In addition, mice were immunized by dropping HA-DNA (12 microg) three times, 2 weeks between each dose into nostrils where each of two electrode needles was placed on the right nostril or the palate. One week after the immunization, the mice were infected with a sublethal dose of the PR8 virus. The DNA immunization by electroporation provided reduced nasal virus titers, in parallel with a relatively high levels of serum anti-HA IgG Ab and a slight nasal anti-HA IgA Ab production. The intranasal administration of cholera toxin before HA-DNA immunization by electroporation enhanced the nasal IgA Ab production together with enhancement of the efficiency of protection. These results suggest that electroporation can be used as one of the efficient gene delivery systems for the transfer of influenza DNA-vaccine into muscle or nasal mucosa to provide protection against influenza virus infection. PMID- 10812220 TI - Minimization of chronic plasma viremia in rhesus macaques immunized with synthetic HIV-1 Tat peptides and infected with a chimeric simian/human immunodeficiency virus (SHIV33). AB - HIV-1 Tat protein activates resting cells, rendering them permissive for viral replication. Replication of HIV-1 in vitro is enhanced by intercellular passage of Tat protein and inhibited by anti-Tat antibodies. Tat dependence of HIV-1 replication in vivo during acute, chronic asymptomatic and AIDS stages of infection was assessed by comparisons of plasma viremia in Tat-immunized or control monkeys challenged with SHIV(33) or SHIV(33A). Chronic plasma viremia became undetectable or minimized in Tat-immunized asymptomatic SHIV(33)-infected monkeys (p<0.008) while the high viral loads of acute infection or SHIV(33A) induced simian AIDS were unaffected by Tat immunization. Active or passive immunotherapies targeting Tat provide potential approaches to controlling chronic HIV-1 viremia and preventing AIDS. PMID- 10812221 TI - Reactogenicity and immunogenicity of a new combined measles-mumps-rubella vaccine: results of a multicentre trial. The Cooperative Group for the Study of MMR vaccines. AB - A large single blind, multi-centre study involving 1779 children was performed in Italy. Infants, aged between 12 and 27 months were divided between two groups: group A received a single dose of a new MMR vaccine, 'Priorix'(3), while group B received a widely used MMR vaccine, Triviraten(4). Solicited local and general symptoms were recorded using diary cards and antibody levels were measured, prior to and 60 days post-vaccination, using ELISA assays. The incidence of solicited symptoms (evaluated in 1754 subjects) was comparable between groups, with the exception of fever which was significantly lower in group B. Immunogenicity was evaluated in 686 subjects. Of note, was the significantly higher anti-mumps seroconversion rate (p<0.001) observed in group A (97.0%) compared to group B (35.4%). However the anti-measles and anti-rubella seroconversion rates were equivalent between groups. Significantly higher (p<0.001) post-vaccination GMTs were in group A vs group B for anti-measles (2830 vs 784 IU/ml) and anti-mumps (1640 vs 469 U/ml), however the anti-rubella GMTs were significantly higher (p<0.001) in group B (117.6 IU/ml) compared to group A (92.6 IU/ml). The persistence of antibodies in 35 subjects was assessed 1 year after vaccination and the results showed no appreciable decline in titres with either vaccine. The trial demonstrates 'Priorix' is well tolerated and highly immunogenic. PMID- 10812222 TI - Adult dogs receiving a rabies booster dose with a recombinant adenovirus expressing rabies virus glycoprotein develop high titers of neutralizing antibodies. AB - Retired greyhound dogs, with low or absent antibody titers to rabies virus following previous vaccinations with commercially available vaccines, were immunized either subcutaneously or intramuscularly with a replication-defective recombinant adenovirus expressing the rabies virus glycoprotein termed Adrab.gp. Immunized animals developed high titers (geometric mean titers of 2630 and 5329) of viral neutralizing antibodies (VNA) against rabies virus by 10 days after vaccination. The antibody titers were even higher (geometric mean titers of 19349 and 122086) by 21 days after vaccination. The results indicate that the recombinant adenovirus expressing rabies virus glycoprotein is capable of inducing antibody immune responses in dogs and therefore may be developed as a rabies virus vaccine for dogs. PMID- 10812223 TI - Effect of individual conjugate dose on immunogenicity of type 6B pneumococcal polysaccharide-N. meningitidis outer membrane protein complex conjugate vaccines in infant rhesus monkeys. AB - A pneumococcal conjugate vaccine (PCV) has been developed consisting of capsular polysaccharide (Ps) coupled to the outer membrane protein complex of Neiserria meningitidis serogroup B. Experiments were conducted in infant rhesus monkeys to assess the potential to administer multiple Pn types in a single vaccine. A single type conjugate, 6B, was dosed from 0.025 to 25 microg Ps. Peak anti-6B Ps Ab titers were seen at lower doses of 0.025 and 0.25 microg Ps, while reduced titers of anti-6B Ps Ab were observed at the highest doses of conjugate administered, 2.5 and 25 microg Ps. By mixing free Ps, carrier, or another monovalent PCV with this 6B PCV, it was determined that reduced anti-6B Ps titers at high PCV doses were associated only with the quantity of type-specific Ps in the conjugate. Thus, increasing the amount of carrier protein or adding an additional monovalent conjugate did not significantly affect the response to type 6B Ps. These results suggest that, given an appropriately determined dose per individual pneumococcal Ps type, a multivalent PCV that includes many different types should have satisfactory clinical immunogenicity. PMID- 10812224 TI - Nanogram quantities of a DNA vaccine protect rainbow trout fry against heterologous strains of infectious hematopoietic necrosis virus. AB - The efficacy of a DNA vaccine containing the glycoprotein gene of infectious hematopoietic necrosis virus (IHNV), a rhabdovirus affecting trout and salmon, was investigated. The minimal dose of vaccine required, the protection against heterologous strains, and the titers of neutralizing antibodies produced were used to evaluate the potential of the vaccine as a control pharmaceutical. Results indicated that a single dose of as little as 1-10 ng of vaccine protected rainbow trout fry against waterborne challenge by IHNV. An optimal dose of 100 ng per fish was selected to assure strong protection under various conditions. Neutralizing antibody titers were detected in fish vaccinated with concentrations of DNA ranging from 5 to 0.01 microg. Furthermore, the DNA vaccine protected fish against a broad range of viral strains from different geographic locations, including isolates from France and Japan, suggesting that the vaccine could be used worldwide. A single dose of this DNA vaccine induced protection in fish at a lower dose than is usually reported in mammalian DNA vaccine studies. PMID- 10812225 TI - Cytological and immunological changes in bronchoalveolar lavage fluid and histological observation of lung lesions in pigs immunized with Mycoplasma hyopneumoniae inactivated vaccine prepared from broth culture supernate. AB - We have compared the cytology of bronchoalveolar lavage fluid (BALF) and the pathology of lung lesions in pigs immunized with/without Mycoplasma hyopneumoniae inactivated vaccine prepared from broth culture supernate on experimental infection. Numbers of total cells, macrophages, neutrophils and lymphocytes have decreased in BALF of vaccinated pigs following infection. The mean percentage of lung lesions, inflammatory cell infiltration into the airways and T cells accumulation around the bronchi were reduced in vaccinated pigs. The levels of tumor necrosis factor (TNF)-alpha also decreased in vaccinated pigs. These results suggest that the vaccination may contribute to decrease TNF-alpha production, and therefore, inflammatory cell responses in the lung due to M. hyopneumoniae infection were suppressed, resulting in fewer lung lesions. PMID- 10812226 TI - Injection with interleukin-4-secreting fibroblasts efficiently induces T helper type 2 cell-dominated immune response. AB - To determine whether the paracrine secretion of interleukin-4 (IL-4) can efficiently induce T helper type 2 (Th2) cell-dominated immune response, BLK fibroblasts were stably transfected to secrete IL-4 (750 units/10(6) cells/48 h). Their effects on T helper cell-mediated immune response were investigated in ovalbumin (OVA)-primed C57BL/6 mice, and were compared with those of free recombinant IL-4. Injection with IL-4-secreting fibroblasts (BLK/IL-4) significantly increased anti-OVA IgG1 production in OVA-primed mice. In addition, the BLK/IL-4 cells were more effective than free recombinant IL-4 in decreasing OVA-specific IFN-gamma production and in increasing OVA-specific IL-4 production by splenic CD4(+) T cells. This work suggests that IL-4-secreting fibroblasts can efficiently induce Th2 cell-dominated immune response and may be beneficial in the treatment of diseases caused by undesired Th1 cell-dominated responses. PMID- 10812227 TI - Immunopotentiating of mucosal and systemic antibody responses in mice by intranasal immunization with HLT-combined influenza virosomal vaccine. AB - The mucosal vaccination strategy against influenza has been investigated by using influenza virosomal vaccine (IRIV) combined with two different adjuvants: the procholeragenoid (PCG) and the Escherichia coli heat labile toxin (HLT). A comparative study has been carried out on mice administered intranasally with these different formulations of influenza vaccine. PCG appears less effective than HLT in inducing an IgG response, but both the adjuvants elicit mucosal adjuvant activity inducing s-IgA in the upper respiratory tract. On the contrary, only HLT when administered intranasally to mice with influenza virosomes stimulates the production of s-IgA in the lower respiratory tract thereby providing a better protection against primary infection of the respiratory system. Both HLT and PCG enhance the production of IFN-gamma in the respiratory tract, nevertheless HLT appears more efficacious as a mucosal adjuvant. PMID- 10812228 TI - High immunogenicity in chimpanzees of peptides and lipopeptides derived from four new Plasmodium falciparum pre-erythrocytic molecules. AB - We have investigated the immunogenicity in chimpanzees of twelve synthetic peptides derived from four new Plasmodium falciparum molecules expressed at pre erythrocytic stages of the human malaria parasite. These parasite molecules were initially selected through their ability to be recognized by stage restricted human antibodies. Twelve 20- to 41-mer peptides representing potential human B- or T-cell epitopes were selected from these proteins, and synthesized. Six of these were modified by a C-terminal lipidic chain in order to re-inforce their immunogenicity. Strong B- and T-helper cell responses were induced in chimpanzees by lipopeptides injected without adjuvant and by peptides in Montanide. All twelve peptides induced CD4(+) T-cell proliferative responses, as well as the secretion of IFN-gamma (some of them at very high levels) and eleven peptides induced antibody responses. The immune responses elicited in this way were reactive with native parasite proteins, as shown by recall studies with sporozoite stage proteins, and proved to be long-lasting (up to 10 months after immunization). Our results support the strategy employed to select these four new malarial antigens and the corresponding peptides, and suggest that the immunizing formulations are both efficient and clinically acceptable. PMID- 10812229 TI - Isolation, purification and immunological characterization of novel low molecular weight protein antigen CFP 6 from culture filtrate of M. tuberculosis. AB - A novel immunogenic antigen, CFP 6 was purified from culture filtrate of M. tuberculosis by a preparatory 2-D electrophoresis method. The protein focused at pI of 4.0 during isoelectric focusing. Molecular weight of the purified protein by ES MS was found to be 11.61 kD. N-terminal amino acid sequence of CFP 6 could be aligned to the deduced amino acid sequence from ORF Rv3004 and was found to be a novel protein with 112 aa residues. Single N-terminal sequence showed that the purified protein was essentially free from contaminants and the amino acid analysis of the antigen was in good agreement with the DNA sequence deduced amino acid composition. Purified CFP 6 was studied for its ability to induce proliferative responses of peripheral blood lymphocytes from five categories of human subjects. These were: untreated, active pulmonary tuberculosis patients; patients after 2-3 months of chemotherapy; vaccinated professional contacts; vaccinated/nonvaccinated healthy controls. CFP 6 elicited high proliferative responses in healthy contacts and patients recovering from the disease. This protein also induced the release of a significantly high amount of IFN-gamma in cell culture supernatant of healthy contacts as compared to other categories of subjects. This protein was further evaluated and compared with PPD and total CS for its DTH inducing ability in guinea pigs immunised with BCG or M. tuberculosis H(37)Rv. CFP 6 elicited a powerful immune response in vitro and in vivo animal model, hence seems to be an immunologically important protein. PMID- 10812230 TI - A genetically-defined riboflavin auxotroph of Actinobacillus pleuropneumoniae as a live attenuated vaccine. AB - Actinobacillus pleuropneumoniae is a gram negative pleiomorphic rod that is the causative agent of a severe, highly infectious and often fatal pleuropneumonia in swine. We have previously reported the construction of genetically-defined stable riboflavin auxotrophs by replacement of a portion of the APP riboflavin biosynthetic operon (ribGBAH) with an antibiotic cassette encoding resistance to kanamycin, and have demonstrated that such riboflavin auxotrophs are avirulent. In this study, we evaluated riboflavin auxotrophs of A. pleuropneumoniae for their ability to stimulate protective immunity against pleuropneumonia. An initial challenge experiment demonstrated that intramuscular vaccination with a live attenuated serotype 1A rib mutant, in a vaccine formulation that included a limiting amount of exogenous riboflavin, provided better protection against challenge with virulent A. pleuropneumoniae than either intratracheal immunization or intramuscular immunization with live bacteria in the absence of exogenous riboflavin. Subsequent studies in which the vaccine inoculating dose, concentration of exogenous riboflavin, and serotype of the vaccine strain were varied demonstrated that immunization with live avirulent riboflavin auxotrophs could elicit significant protection against experimental challenge with both homologous and heterologous virulent serotypes of A. pleuropneumoniae. PMID- 10812231 TI - Recombinant polioviruses expressing hepatitis B virus-specific cytotoxic T lymphocyte epitopes. AB - T-cell immune response to recombinant poliovirus expressing foreign antigens has not been elucidated well. In order to investigate the potential use of poliovirus as a T-cell vaccine vector, we constructed recombinant polioviruses expressing HBV-derived CTL epitopes, HBsAg(28-39) (L(d)-restricted; IPQSLDSWWTSL) and HBc(93 100) (K(b)-restricted; MGLKFRQL) at the junction between the P2 and P3 regions, designated V3CDs and V3CDc, respectively. The V3CDs and V3CDc recombinant viruses replicated efficiently in HeLa cells and showed a similar infection profile to that of the wild-type Mahoney strain in one-step growth kinetics. Genetic stability analysis showed that V3CDc retained the foreign insert over twelve successive passages examined, whereas V3CDs lost part of the foreign insert after four passages. Our results indicated that the stability of the inserted foreign sequences was rather affected by their nucleotide sequence than by their length when located between the P2 and P3 regions. The junction between these nonstructural protein-coding regions is a novel site for the construction of replication-competent recombinant poliovirus. Immunization of BALB/c (H-2(d)) and C57BL/6 mice (H-2(b)) with V3CDs and V3CDc, respectively, elicited significant antigen-specific CTL responses to HBsAg(28-39) but not to HBcAg(93-100). PMID- 10812232 TI - Augmentation of antitumor immunity by genetically engineered fibroblast cells to express both B7.1 and interleukin-7. AB - Mouse fibroblasts (H-2(b)) were genetically engineered to express a costimulatory B7.1 and an interleukin-7 (IL-7; Fb/B7.1/IL7). The Fb/B7.1/IL7 cells were then pulsed with an ovalbumin (OVA) epitope (amino acids 257-264, SIINFEKL, H-2 K(b) restricted; Fb/B7. 1/IL7/OVA) and tested for the induction of OVA-specific cytotoxic T lymphocytes (CTLs) in C57BL/6 mice (H-2(b)). The genetically engineered fibroblasts lacking either B7.1 or IL-7 were constructed and used as controls. Immunization with the Fb/B7.1/IL7/OVA cells induced strong cytotoxic activities against OVA-expressing EL4 (EG7) tumor cells. The magnitude of the cytotoxic response in mice with the Fb/B7.1/IL7/OVA cells was significantly higher than the response in mice immunized with any other cell constructs. CD8(+) T cells were a major effector cell-type of antitumor response in the immunized mice with the Fb/B7.1/IL7/OVA cells. Furthermore, immunization with the Fb/B7.1/IL7/OVA cells significantly prolonged the survival period of mice when the mice were injected with EG7 tumor cells one week after the immunization. These results suggest that fibroblasts can be genetically modified to an efficient cell vaccine for the induction of antitumor response. PMID- 10812233 TI - Competition among Streptococcus pneumoniae for intranasal colonization in a mouse model. AB - Widespread use of conjugate vaccines against Streptococcus pneumoniae, by reducing carriage of S. pneumoniae serotypes included in the vaccine, may result in an increase in nasopharyngeal carriage of - and disease from - nonvaccine serotypes of the same species. Mathematical models predict that the extent of such replacement will depend positively on the degree to which carriage of vaccine-type S. pneumoniae inhibits acquisition of nonvaccine-type pneumococci, and may depend negatively on the inhibition of vaccine-type pneumococci by nonvaccine-type pneumococci. We used a mouse model of intranasal carriage of pneumococci to test whether such inhibition occurs between different pneumococcal strains. Mice carrying a streptomycin-resistant derivative of S. pneumoniae BG9163 (serotype 6B) as a resident strain showed reduced levels of colonization when challenged intranasally by optochin-resistant derivatives of the same strain and of a serotype 23F pneumococcus, BG8826. Inhibition could be overcome by increasing the dose of the challenge strain. Carriage of optochin-resistant BG9163 did not inhibit acquisition of the streptomycin-resistant variant. Colonization by a challenge strain did not significantly affect the level of colonization with the resident strain. These results provide evidence that is consistent with several hitherto untested assumptions of mathematical models of serotype replacement and suggest that a biological mechanism exists that could account for serotype replacement that is observed in clinical trials. The findings provide a basis for further studies of in vivo interactions between strains of S. pneumoniae. PMID- 10812234 TI - Development, expression, and murine testing of a multistage Plasmodium falciparum malaria vaccine candidate. AB - A synthetic gene encoding twelve B cell epitopes, six T-cell proliferative epitopes, and three cytotoxic T lymphocyte (CTL) epitopes from nine stage specific antigens, representing the sporozoite, liver stage, asexual blood-stage, and sexual-stage antigens of Plasmodium falciparum, was constructed by assembling overlapping oligonucleotides followed by PCR extension and annealing. A three step PCR protocol using twelve long oligonucleotides was employed to generate a 1053 base-pair synthetic gene, the identity of which was confirmed by sequencing. This synthetic gene, named CDC/NII MAL VAC-1, was cloned, and the recombinant protein was expressed in the Baculovirus Expression Vector System (BEVS). The selection of malarial epitopes for inclusion in this vaccine construct was based on immunoepidemiological studies in malaria endemic area, in vitro, and in vivo protection studies in model systems. The 41 kDa BEVS-expressed recombinant protein reacted with mouse antibodies specific for individual B cell epitopes in the vaccine construct and with sera from clinically immune Kenyan adults. An immunization study in three strains of mice that differ at the H-2 locus demonstrated that the BEVS-expressed recombinant protein is immunogenic; the candidate vaccine antigen induced high titer antibodies, and lymphocyte proliferative and IFN-gamma responses. These results demonstrate that individual B and T cell epitopes can be assembled to create synthetic genes that encode proteins capable of eliciting specific antibody and T cell responses. PMID- 10812235 TI - Comparison of a live attenuated and an inactivated varicella vaccine to boost the varicella-specific immune response in seropositive people 55 years of age and older. AB - Healthy, varicella-zoster virus (VZV)-seropositive subjects, aged 55-89 years (mean age 66 years), received either 4000 PFU of live, attenuated VZV vaccine (n=85) or an equal volume of this vaccine that was heat-inactivated (n=82). Both vaccines significantly boosted VZV antibody (enzyme immunoassay) and gamma interferon production by peripheral blood mononuclear cells stimulated by VZV antigen. These responses returned to baseline by 12 months. Circulating mononuclear cells that proliferated in response to VZV antigen were significantly more numerous (responder cell frequency assay) after either vaccine, and persisted with a half-life of 17. 5-21.3 months. There were no differences in immune response to either vaccine in this older age cohort. PMID- 10812236 TI - Immunogenicity of Ty-VLP bearing a CD8(+) T cell epitope of the CS protein of P. yoelii: enhanced memory response by boosting with recombinant vaccinia virus PMID- 10812237 TI - Tolerance and hematopoietic stem cell transplantation 50 years after Burnet's theory. AB - OBJECTIVE: In 1949, the original formulation of Burnet's theory on the mechanisms responsible for the capacity of the immune system to discriminate between foreign antigens (i.e., the "non-self") and the cells of its own body (i.e., the "self") was published. Since then, further refinements and reconsiderations of the basic concepts underlying the achievement of a state of tolerance toward a certain antigen have been reported. Here, we attempt to analyze critically new clinical and experimental strategies aimed at inducing alloantigen-specific unresponsiveness. DATA SOURCES: The data discussed in this review are drawn from articles and abstracts published in journals covered by the Science Citation Index and Medline. STATE OF THE ART: Induction of tolerance toward alloantigens still remains one of the most elusive goals of clinical immunology. Until now, nonspecific immunosuppressive drugs have been used to successfully perform both solid organ and hematopoietic stem cell transplantation. However, using this approach, patients given an allograft are exposed to the threat of infections, tumors, and other side effects. Moreover, in solid organ transplant recipients, permanent tolerance toward the graft's alloantigens is never achieved. Recently, considerable progress has been made in expanding our knowledge of transplant tolerance. The traditional model of central tolerance, derived from Burnet's concept, has been complemented by knowledge of mechanisms of peripheral tolerance. CONCLUSIONS: New experimental and therapeutic trials based on the blockade of costimulatory molecules, as well as on generation and infusion of either regulatory or nonimmunogenic cells, have been recently proposed for inducing alloantigen-specific tolerance.The achievements obtained in understanding the mechanisms of unresponsiveness toward non-self antigens are fundamental prerequisites for successful allogeneic transplants, and they could open a new exciting era of specific, immunosuppressive therapies. PMID- 10812238 TI - The fusion protein MEN 11303 (granulocyte-macrophage colony stimulating factor/erythropoietin) acts as a potent inducer of erythropoiesis. AB - OBJECTIVE: A fusion protein made of human granulocyte-macrophage colony stimulating factor (GM-CSF) and erythropoietin (EPO), referred to as MEN 11303, has been tested for biologic activity using mobilized CD34(+) cells. METHODS AND RESULTS: MEN 11303 and a combination of GM-CSF/EPO produced the same amount of colony-forming unit granulocyte-macrophage (CFU-GM), of burst-forming unit erythroid (BFU-E), and of multipotent CFU-mixed. After 15 days, liquid cultures of CD34(+) cells exposed to MEN 11303 yielded a total cell number larger than that obtained with an equimolar mixture of GM-CSF and EPO, with a clear prevalence of cells exhibiting an erythroid phenotype. A colony-forming cell assay established from CD34(+) cells precultured with MEN 11303 for 7 days yielded a greater amount of BFU-E than GM-CSF/EPO combination. Exposing CD34(+) cells to MEN 11303 for 7 days in liquid culture resulted in higher recoveries of cells expressing a comparatively less differentiated hematopoietic phenotype and of long-term culture initiating cells. A cell-based binding-competition assay using the human EPO-receptor (EPO-R) transfected murine Ba/F3EPOR cell line showed that MEN 11303 bound to EPO-R with a sixfold lower affinity but induced a more sustained receptor phosphorylation. MEN 11303 supported the growth of Ba/F3EPOR cells more efficiently than EPO and remained detectable in the spent culture medium for a longer time. CONCLUSIONS: MEN 11303 and the combination of GM-CSF/EPO are equally potent in recruiting hematopoietic progenitors into cycle, but the fusion protein is superior in promoting the expansion of committed erythroid percursors. Primitive hematopoiesis is less affected by MEN110303 than GM-CSF/EPO combination. Part of these effects may reflect the peculiar interaction of the EPO moiety of MEN 11303 with the EPO-R. PMID- 10812239 TI - Immune dysfunction despite high levels of immunoregulatory cytokine gene expression in autologous peripheral blood stem cell transplanted non-Hodgkin's lymphoma patients. AB - OBJECTIVE: In the present studies, we examined the role of immunoregulatory cytokine gene expression in immune reconstitution following high-dose chemotherapy and peripheral stem cell transplantation. MATERIALS AND METHODS: We analyzed the steady-state mRNA cytokine levels and the immune phenotype and function in the peripheral blood mononuclear cells from intermediate-grade non Hodgkin's lymphoma patients prior to and following high-dose chemotherapy and peripheral stem cell transplantation. RESULTS: Significantly higher mRNA levels of both type 1 and type 2 cytokines and monokines were observed in patients undergoing high-dose chemotherapy and peripheral stem cell transplantation as compared with normal healthy individuals. Pretransplant mRNA levels of interleukin-2, -4, -8, -10, interferon-gamma and tumor necrosis factor-alpha were significantly higher than in normal individuals. In addition, on days 30 and 100 following transplantation interleukin-10 levels were significantly increased compared with pretreatment levels. In contrast, the levels of interleukin-2 mRNA and interferon-gamma were decreased significantly on day 365 compared with pretransplant levels. CONCLUSIONS: The high levels of cytokine mRNA transcripts, both prior to and following peripheral stem cell transplantation, were not due to an increased cellular frequency; rather, they appear to be due to abnormal cellular activation. However, T-cell function is significantly depressed compared with normal donors, which is associated with significantly higher levels of cellular-dependent T cell inhibitory activity and, we suggest herein, high levels of interleukin-10, a type 2 cytokine. PMID- 10812240 TI - Differential engraftment of genetically modified CD34(+) and CD34(-) hematopoietic cell subsets in lethally irradiated baboons. AB - OBJECTIVE: To test gibbon ape leukemia virus (GALV) pseudotype vector transduction of marrow subpopulations that contribute to hematopoietic reconstitution in vivo. MATERIALS AND METHODS: Autologous CD34(+) Lin(-), CD34(+) Lin(+), and CD34(-) Lin(-) marrow cells, transduced by coculture with PG13/LN, PG13/LNX, and PG13/LNY vector-producing cells, respectively, were transplanted in three female baboons. Two female baboons also were transplanted with fresh allogeneic CD34(-)Lin(-) marrow cells from MHC-matched male siblings and, to ensure survival, with autologous CD34(+)Lin(-) and CD34(+)Lin(+) marrow cells transduced with PG13/LN and PG13/LNX, respectively. The LN, LNX, and LNY vectors are identical except for different length sequences at the 3' end of the bacterial neomycin phosphotransferase (neo) gene. RESULTS: LN(+) and LNX(+) cells from CD34(+)Lin(-) and CD34(+)Lin(+) cells, respectively, but no LNY(+) from CD34(-)Lin(-) cells were detectable in blood and marrow of all animals after transplant. LN(+), CD34(+)Lin(-) cells contributed to reconstitution of the T, B, and myeloid lineages. LNX(+), CD34(+)Lin(+) cells contributed only to B and myeloid lineages. Male cells, CD34(-)Lin(-), were detected by polymerase chain reaction in blood and marrow of the two allogeneic transplanted animals at estimated frequencies of 20% total burned surface area (TBSA), at admission and 24 h post-admission.Eleven patients (mean age 46.5 years, 8 males) with a mean TBSA of 34.5% were sampled. Standard treatment was given. Eleven closely age-matched volunteers were used as controls. A specific ELISA method suitable for the measurement of ANP and VIP was used.ANP was higher (p<0.0001), while VIP was lower (p=NS) in patients' samples compared to controls. While the level of VIP was higher at 24 h post-admission, mean ANP level remained about the same. The increased levels of plasma ANP may result from volaemic disturbances during resuscitation, low VIP levels, the increase in pulmonary resistance or post-burn stress. PMID- 10812267 TI - Changes in circulating levels of an anti-inflammatory cytokine interleukin 10 in burned patients. AB - In order to understand the role of an anti-inflammatory cytokine interleukin 10 (IL-10) in the pathophysiology of burn injury, IL-10 levels in serial serum samples of 22 burned patients were analyzed. The total body surface areas (TBSA) of the burn injury ranged from 30 to 90%. Among these 22 patients, 14 recovered and the other eight, who were septic, expired. A significant difference in serum IL-10 values on admission (5-20 h postburn) was found (P<0.05) between patients who survived or died from burn injury as analyzed by the Student's t test. In addition, a significant difference in serum IL-10 on admission was also found (P<0.05) between patients with TBSA of greater or less than 50%. An initial peak serum IL-10 response was detected within 2.5 days postburn. Significant differences in the peak serum IL-10 levels were not found between patients with TBSA of greater or less than 50% and patients who survived or expired from burn injury. Afterwards, serum IL-10 remained low in the survivors, while an increase in serum IL-10 could be detected in the non-survivors with proven sepsis. Levels of circulating IL-6 in these non-surviving patients showed a tendency to increase starting from about 1-2 weeks postburn which coincided temporally with the detection of infections. However, marked increases in circulating IL-10 levels were observed just before death in four of the eight non-survivors. The serum samples of these four patients were collected at 31 h (404.8 pg/ml), 2 h (773.9 pg/ml), 5 days (150.7 pg/ml) and 12 h (177.1 pg/ml) before the expiration of these patients, respectively. IL-10 levels of 28.6, 27. 5 and 13.5 pg/ml were detected in sera of three of the remaining four non-survivors that were collected at 2.5 h, 36 h and 30 h before the expiration of these patients, respectively. There was one non-surviving patient who suffered an 80% burn (patient D4 in Table 1 and Fig. 4) and his IL-10 level at 20 days postburn was 13.4 pg/ml. The serum sample of this patient was collected 22 days before death and he was not suffering from sepsis at this stage. In conclusion, an initial increase in serum levels of IL-10 was detected postburn. A marked increase in serum levels of IL-10 was detected in four of the eight septic patients just before their death. It was considered that a lack and/or a delay in the increase of circulating IL-10 may play a significant role in the pathophysiology of sepsis in burned patients. PMID- 10812268 TI - A prospective study of suicidal burns admitted to the Harare burns unit. AB - The aim of this study was to obtain prospective information on suicidal (attempted suicide) burns patients admitted to the Harare burns unit during 1995 1998. Forty-seven patients, 42 females (89%) and five males (11%), evenly distributed throughout the period of study, were included. The median age was 25 years, range 13 to 50 years. Thirty were housewives (64%). Women married according to customary law were the group most at risk. All patients were burnt by flame after dousing themselves with paraffin or petrol. Conflict in love relationships was the most common circumstance leading to attempted suicide. The median Total Body Surface Area (TBSA) burnt was 60%, range 10-90%, for all patients, 25%, range 10-40%, for those who survived and 65%, range 20-95%, for those who died. Surgery was performed on 16 patients (34%). Mortality was 68%. The overall median hospital stay for all patients was 10 days, range 0-322 days, and 5 days, range 0-322 days, for those who died. PMID- 10812269 TI - Epidemiology of childhood burns in the critical care medical center of Kinki University Hospital in Osaka, Japan. AB - The objective of the present study was to describe the characteristics of pediatric burns in order to prepare a program for the prevention of severe burn injuries in children. We conducted a retrospective study of burn victims aged 15 years or younger who were hospitalized in our Critical Care Medical Center between 1982 and 1997. There were 73 children with burn injuries hospitalized in our center during the study period. The greatest number were children 1 year old. The average % body surface area burned was 21. 5+/-20.5%. The most important causes of pediatric burns were found to be hot bath water and other hot liquids. Hot bath scalds accounted for about half of the pediatric burns occurring in all age groups, and they were often extensive. Non-bath scalds accounted for about one-third of the pediatric burns and were most frequent in children 2 years and younger. All the injuries sustained at home occurred when a family member was in the house. Similar to many reports from overseas, non-bath scalds were one of the most common causes of burns in this study; however, hot bath scalds were the most important cause. These data are being used to develop a prevention program. We also consider it necessary to educate children and their family members about the dangers of burn injuries. PMID- 10812270 TI - "Train surfers": analysis of 23 cases of electrical burns caused by high tension railway overhead cables. AB - The term "train surfers" describes a group of adolescents from the outskirts of the city of Rio de Janeiro, who are compelled by the overcrowded railway trains to travel on the roofs of the wagons. Collision with electrical high-tension wires is a relatively frequent occurrence, causing extensive and complex injuries. This study analyzes this clinical and surgical phenomenon which has caused over 100 fatalities in more than 200 registered accidents over the past 10 years. PMID- 10812271 TI - Kerosene stove as a cause of burns admitted to the Ain Shams burn unit. AB - Kerosene stoves are in widespread use for cooking and warming water in underprivileged areas in Cairo. These stoves are dangerous and lack safety measures; they are often a cause of fire incidents and burn injuries. During the period from May 1995 to December 1996 the number of patients who presented to the burn unit of Ain Shams University, Cairo, Egypt was 759, of whom 304 (40%) sustained the injury as a consequence of kerosene stove fires. Efforts to inform the public about the danger of these stoves are recommended to minimize the incidence, morbidity, mortality and cost of this relatively common and preventable type of injury. PMID- 10812272 TI - Aeromonas wound infection in burns. AB - Infection of burn patients with the Aeromonas organism is an uncommon event. This paper documents four cases of Aeromonas hydrophila and one case involving both A. hydrophila and A. caviae occurring in burn patients between 1990 and 1998 at the Royal Brisbane Hospital burns unit. The organism was isolated from either skin swabs, tissue samples, blood cultures or cultured lines. In all patients there was a history of immersion in water immediately post burn. There is one case of invasion and destruction of deeper tissues and one fatality. Appropriate management requires a high index of suspicion if a history of immersion in untreated water post burn is given and the treatment involves aggressive excision and antibiotic therapy. PMID- 10812274 TI - Osteomyelitis in burn patients requiring skeletal fixation. AB - Deep and severe burns often present with the exposure of musculoskeletal structures and severe deformities. Skeletal fixation, suspension and/or traction are part of their comprehensive treatment. Several factors put burn patients at risk for osteomyelitis, osteosynthesis material being one of them. In order to determine the safety of skeletal fixation, we reviewed all pediatric patients treated with pin insertion during the past 10 years. Forty-one severely burned children (61+/-3% TBSA full thickness burns), had a total of 357 skeletal fixation procedures. Pins were maintained an average of 25.3+/-1.7 days. Thirteen pins (3. 6%) were loose before the expected time of removal, two patients (4. 8%) presented with cellulitis of the pin site and two patients (4. 8%) presented with osteomyelitis. Loose pins, pin site cellulitis, burn wound infection and sepsis were not associated with osteomyelitis. Skeletal fixation in severely burned patients presents with a low incidence of infectious complications. Its use should be considered in patients affected by severe burns that present with exposed deep structures and for positioning purposes. The exact timing for pin removal in burned patients is still to be defined. PMID- 10812273 TI - Trichosporon beigelii infection: experience in a regional burn center. AB - Trichosporon beigelii is a fungus once thought to cause only superficial infections, but recently has been increasingly identified as an opportunistic systemic pathogen in immunocompromised patients. There have been very limited reports of this organism in the burn patient population. We describe the first report of pharmacological management of invasive T. beigelii with a combination of amphotericin B and high dose fluconazole in a burn patient. Antifungal susceptibility testing of T. beigelii determined a change in minimum inhibitory concentrations (MICs) of amphotericin B and a consistent resistance pattern with the use of flucytosine. This paper will review our experience with T. beigelii fungus in a regional burn treatment center and review the literature on other experiences in the burn population. PMID- 10812275 TI - A near closed book contracture of the ear: a case report. AB - An unusual post-burn ear contracture, involving crus helicis, cymba conchae, tragus and antihelix, with anterior vertical folding of the ear on itself, like a closed book, has been described. The contracture was completely released and the resultant raw area was covered by a full thickness skin graft, harvested from the opposite post-auricular area. The crippled ear cartilage was still folded anteriorly at the level of the concha, regardless of complete release, due to long standing childhood burns. Three months later, concha-mastoid suturing was carried out to give the ear its normal shape and erect, prominent position. Four years of follow up showed, an aesthetically good-looking ear with satisfactory position. PMID- 10812276 TI - Anhydrous ammonia burns case report and review of the literature. AB - Chemical burns are associated with significant morbidity, especially anhydrous ammonia burns. Anhydrous ammonia is a colorless, pungent gas that is stored and transported under pressure in liquid form. A 28 year-old patient suffered 45% total body surface area of second and third degree burns as well as inhalational injury from an anhydrous ammonia explosion. Along with fluid resuscitation, the patient's body was scrubbed every 6 h with sterile water for the first 48 h to decrease the skin pH from 10 to 6-8. He subsequently underwent a total of seven wound debridements; initially with allograft and then autograft. On post burn day 45, he was discharged. The injuries associated with anhydrous ammonia burns are specific to the effects of ammonium hydroxide. Severity of symptoms and tissue damage produced is directly related to the concentration of hydroxyl ions. Liquefactive necrosis results in superficial to full-thickness tissue loss. The affinity of anhydrous ammonia and its byproducts for mucous membranes can result in hemoptysis, pharyngitis, pulmonary edema, and bronchiectasis. Ocular sequelae include iritis, glaucoma, cataracts, and retinal atrophy. The desirability of treating anhydrous ammonia burns immediately cannot be overemphasized. Clothing must be removed quickly, and irrigation with water initiated at the scene and continued for the first 24 h. Resuscitative measures should be started as well as early debridement of nonviable skin. Patients with significant facial or pharyngeal burns should be intubated, and the eyes irrigated until a conjunctivae sac pH below 8.5 is achieved. Although health care professionals need to be prepared to treat chemical burns, educating the public, especially those workers in the agricultural and industrial setting, should be the first line of prevention. PMID- 10812277 TI - Zinc burns: a rare burn injury. AB - A patient was presented with significant burns resulting from a workplace accident in a zinc production unit. This occurred as a result of the spontaneous combustion of zinc bleed under high pressure. The patient sustained burns to the face, body, and hands and suffered significant injury to the left cornea. Computed imaging revealed solid particles in the ethmoid sinus and also in the right nasal fossa, dissecting the right lacrimal duct. Photographic documentation is presented. This injury was potentially preventable and resulted from poor observance of safety procedures. PMID- 10812278 TI - Artificial skin reduces nutritional requirements in a severely burned child. AB - An 11 year old boy had 60% BSA burns excised and artificial skin (Integra(R)) applied. None of the burn wound was grafted. At 3 weeks post burn his serum biochemistry was normal, except for a low serum albumin. Mean energy intake for the first 3 weeks was half his non-burned requirement and mean protein intake was close to his non-burned requirement. This is the first report to show that a reduction in nutritional requirements may be observed using skin substitutes. PMID- 10812279 TI - Bizarre paediatric facial burns. AB - Child abuse and neglect account for a significant number of paediatric burn injuries. It is of great importance because of the high mortality, high frequency of repeated abuse, as well as the physical, psychological and social sequelae that it causes. Burn abuse is often under-recognized and under-reported because it is difficult to define non-accidental injury. On the other hand, false accusation of burn abuse is extremely damaging to the family. Bizarre and unusual burn injuries can be caused by accident and should not automatically be assumed to be deliberate injury. Three boys of age 1-7 years with bizarre facial burns were admitted to the Burns Unit at the Prince of Wales Hospital between February 1995 and July 1999. One was burned by his baby-sitter with hot water steam and the other two were burned by their mothers with hot boiled eggs. The unusual causes of their burns raised the suspicion of child abuse and formal investigations were carried out by the Social Services Department. Detail assessment including a developmental history of the child and the psychosocial assessment of the family revealed that these three boys were burned because of poor medical advice and innocent cultural belief. PMID- 10812280 TI - Ink used as first aid treatment of a scald. AB - We report here a case of a childhood scald injury where immediate first aid consisted of the application of fountain pen ink. Continuing education is required to inform parents of the use of cold water in the immediate first aid treatment of scalds. PMID- 10812281 TI - Remissions from drug dependence: is treatment a prerequisite? AB - The present study investigated the prevalence of untreated drug remissions in a representative sample of respondents who had a lifetime diagnosis of one of five types of illicit drug dependence in the past (cannabis, cocaine/crack, hallucinogens, amphetamines, or heroin) but no diagnosis of drug abuse or dependence in the last year. It was concluded that, among recovered individuals with previous clinically significant drug concerns (as measured using DSM-IV criteria), a substantial proportion had accessed addictions treatment services prior to remission. PMID- 10812282 TI - Baseline prediction of 7-month cocaine abstinence for cocaine dependence patients. AB - A broad range of baseline subject variables was evaluated to identify predictors of 7-month cocaine use for 160 lower socioeconomic cocaine dependent male veteran patients participating in either an intensive 1-month day hospital (DH; n=90) or a 1-month inpatient (INP; n=70) treatment program. The baseline measures included sociodemographic variables, the seven Addiction Severity Index composite scores, cocaine urine toxicology, craving, the SCL-90 total score, and lifetime psychiatric diagnoses. Since a proportion of subjects who reported no use at follow-up had positive urines, both liberal and conservative data estimation strategies were employed for subjects without urine toxicology data at follow-up who had reported no use (21% of subjects). Analyses were done separately for the DH and INP subjects. Under the conservative definition of cocaine abstinence/use, univariate correlations of predictor variables with 7-month cocaine use revealed no statistically significant relationships. Under the liberal definition of cocaine abstinence/use, only one variable, greater severity of alcohol problems at intake predicted cocaine abstinence at outcome. Because of the inability to predict treatment success, originally planned logistic regression analyses were not undertaken. The findings point to the difficulty of predicting long-term outcomes in cocaine dependent patients based on baseline information and to the importance of obtaining objective data on cocaine use. PMID- 10812283 TI - A controlled trial of daily versus thrice-weekly buprenorphine administration for the treatment of opioid dependence. AB - This study was aimed at determining whether thrice-weekly administration of buprenorphine is as effective as daily administration for treating opioid dependence. A total of 60 treatment-seeking opioid addicts were randomly assigned to take buprenorphine tablets sublingually either every day (8 mg) or thrice weekly (16 mg on Mondays and Wednesdays and 24 mg on Fridays) over the course of a 12-week, double-blind, parallel trial. The buprenorphine dosing schedule had no significant effect on treatment retention. The rates of opioid-positive urine tests were significantly higher among those subjects who were given buprenorphine thrice weekly (58.5%) than among those who took it daily (46.6%). An analysis of the completers did not detect a significant effect of buprenorphine dosing schedule. The results obtained in our clinical trials indicate the advisability of daily doses of buprenorphine, at least at the beginning of a maintenance programme. PMID- 10812284 TI - Drinking as a risk factor for sustained smoking. AB - Data from a nationally representative sample of US adults (n=42862) were used to explore the associations between drinking and smoking, on a lifetime and past year basis, and between drinking and smoking cessation for a subsample of past year smokers (n=12586). Smoking cessation was defined as current nonsmoking (as of the time of interview) that had lasted at least 3 months. The proportions of both lifetime and past-year smokers increased with volume of alcohol intake and frequency of heavy drinking and were greater for individuals with DSM-IV alcohol abuse or dependence than for individuals without these disorders. For example, the proportion of past-year smokers rose from 22.5% of lifetime abstainers to 53.0% of heavy drinkers, from 23.8% of those who never drank 5+ drinks on any drinking day to 61.8% of those who drank 5+ drinks weekly or more often and from 27.6% of individuals without past-year DSM-IV alcohol abuse or dependence to 55.5% of those with either of these disorders. The proportion of past-year smokers who had stopped smoking decreased from 7.8% of former drinkers to 4.6% of heavy drinkers, from 7.3% of those who never drank 5+ drinks to 3.4% of those who did so weekly or more often and from 6.7% among individuals without past-year abuse or dependence compared to 4.4% among those with either disorder. In a multivariate analysis that adjusted for background variables and smoking history, average daily ethanol intake was not significantly associated with the odds of smoking cessation, but drinking 5+ drinks at least once a month reduced the odds of cessation by 42%. Having an alcohol use disorder increased the odds of smoking cessation below the age of 30 (an effect that disappeared after adjusting for the interaction between age group and having children in the home), but had an increasingly negative effect on smoking cessation at older ages. PMID- 10812285 TI - Naltrexone does not block the subjective effects of oral Delta(9) tetrahydrocannabinol in humans. AB - Delta(9)-tetrahydrocannabinol (THC) and opioids have many common effects. In addition, some THC effects in laboratory animals can be blocked or attenuated by opioid antagonists. This suggests that opioid systems mediate or modulate some THC effects. To determine whether opioid systems mediate THC effects in humans, the effects of the opioid antagonist naltrexone on subjective responses to THC were examined in 14 marijuana users. Subjects participated in a double-blinded, cross-over design in which each subject received all combinations of naltrexone (0 or 50 mg) and THC (0, 7.5, or 15 mg). THC increased heart rate and self reported drug effects, such as euphoria and marijuana-like effects, and decreased psychomotor performance. Naltrexone increased heart rate and decreased self reported measures of vigor and hunger but did not alter any of the effects of THC. These results suggest that the subjective, physiological, and behavioral effects of THC in humans are not mediated through opioid systems. PMID- 10812286 TI - Antinociceptive, subjective and behavioral effects of smoked marijuana in humans. AB - The purpose of this study was to determine whether marijuana produced dose dependent antinociception in humans and, if so, whether endogenous opiates modulate this effect. A total of five male regular marijuana users participated in three test sessions during which they smoked cigarettes containing 0% (placebo) and 3. 55% Delta(9)-tetrahydrocannabinol (Delta(9)-THC) (active). Each of four controlled smoking bouts per session, spaced at 40-min intervals, consisted of nine puffs from active and placebo cigarettes (three cigarettes, three puffs per cigarette, one puff per min). During successive bouts, participants smoked 0, 3, 6 and 9 (0, 3, 9 and 18 cumulative) puffs from active marijuana cigarettes, with the remainder of puffs from placebo cigarettes. Test sessions were identical, except for naltrexone 0, 50 or 200 mg p.o. (randomized, double-blind) administration 1 h before the first smoking bout on the different days. Before smoking, between smoking bouts and postsmoking, participants completed an assessment battery that included antinociceptive (finger withdrawal from radiant heat stimulation), biological, subjective, observer-rated signs and performance measures. Marijuana produced significant dose-dependent antinociception (increased finger withdrawal latency) and biobehavioral effects. Naltrexone did not significantly influence marijuana dose-effect curves, suggesting no role of endogenous opiates in marijuana-induced antinociception under these conditions. PMID- 10812287 TI - Acute and protracted cocaine abstinence in an outpatient population: a prospective study of mood, sleep and withdrawal symptoms. AB - This study addresses unresolved questions about cocaine withdrawal by prospectively assessing monitored cocaine abstinence over 28 days in a sample of 24 male and female cocaine-dependent outpatients. Based on results from urine drug screens and self-reported substance use, it is likely that these patients were abstinent from cocaine during the assessment period. Abstinence-related symptoms were monitored at 2, 5, 10, 14, 21, and 28 days following last cocaine use. For patients who were known to relapse, assessments began again after the last day of cocaine use. Consistent with findings from inpatient studies of cocaine abstinence, linear improvements in negative affect, low cocaine craving, and increases in cognitive skills were reported over the 28 days. Also consistent with inpatient studies of cocaine withdrawal, a phasic withdrawal syndrome was not observed in this outpatient sample. Unlike inpatient studies, no disturbances in sleep were reported. PMID- 10812288 TI - Combined use of trazodone-naltrexone versus clonidine-naltrexone in rapid withdrawal from methadone treatment. A comparative inpatient study. AB - Trazodone is a non-tricyclic antidepressant drug with specific antagonistic activities at 5-HT(2) and alpha-1 adrenoceptors. We test the efficacy of trazodone (T) compared with clonidine (C) in rapid opiate detoxification (ROD) from methadone after reduction to a daily maintenance dose 0.05) by age, gender, hypertension status, or history of myocardial infarction. In this clinical population, utilizing the 2-min rhythm strip as an indicator of average PVCs per hr had excellent specificity and moderate to low sensitivity across most of the distribution of average PVCs per hr. The use of a short rhythm strip to detect PVCs may be considered useful in epidemiologic investigations of cardiovascular disease and mortality for detecting high frequency PVCs in populations. The use of a short rhythm strip as a screening tool to detect PVCs in clinical practice is not warranted, based on our findings and the existing literature. However, an awareness that PVCs on a 2-min rhythm strip consistently identify high frequency PVCs on 24-hr recordings should be helpful to clinicians. PMID- 10812322 TI - Bias and prevalence effects on kappa viewed in terms of sensitivity and specificity. AB - Paradoxical effects of bias and prevalence on the kappa coefficient are examined using the concepts of sensitivity and specificity. Results that appear paradoxical when viewed as a 2 x 2 table of frequencies do not appear paradoxical when viewed as a pair of sensitivity and specificity measures where each observer is treated as a predictor of the other observer. An adjusted kappa value can be obtained from these sensitivity/specificity measures but simulation studies indicate that it would result in substantial overestimation of reliability when bias or prevalence effects are observed. It is suggested that investigators concentrate on obtaining populations with trait prevalence near 50% rather than searching for statistical indices to rescue or excuse inefficient experiments. PMID- 10812323 TI - Community-based epidemiological study of glucose tolerance in Kin-Chen, Kinmen: support for a new intermediate classification. AB - In this population-based survey, we investigated the prevalence of varying degrees of glucose tolerance among residents of Kin-Chen, Kinmen, as well as the association of glucose tolerance status with potential risk factors for type 2 diabetes and cardiovascular disease (CVD). We focused particularly on subjects with normal 2-h postload glucose level (<7.8 mmol/l) but persistent fasting hyperglycemia (PFH) (5.6-7.8 mmol/l), to examine whether PFH represents an intermediate state between normal glucose tolerance (NGT) and impaired glucose tolerance (IGT). The target population comprised 6346 residents aged 30 years and older. A total of 4354 subjects could be classified into categories of NGT, PFH, IGT, new diabetes, and known diabetes according to medical history, fasting plasma glucose levels, and the results of a 75-g oral glucose tolerance test (OGTT). The potential cardiovascular risk factors assessed included age, obesity (general and central), systolic blood pressure, and fasting levels of insulin, C peptide, triglyceride, cholesterol, and high-density lipoprotein cholesterol (HDL C). The age-standardized prevalences of PFH, IGT, new diabetes, and known diabetes were 2.9%, 3.5%, 4.0%, and 3.0%, respectively. Among nondiabetic subjects, the cardiovascular risk factor profiles worsened with decreasing glucose tolerance, with most values differing significantly among the NGT, PFH, and IGT groups. Subjects with PFH, who would be classified as having NGT according to conventional WHO criteria, had physical and biochemical features between those of the NGT and IGT groups. These findings support our previous observation that PFH may be a transition state between NGT and IGT in the progression toward type 2 diabetes. PMID- 10812324 TI - Diabetes mellitus, coronary heart disease incidence, and death from all causes in African American and European American women: The NHANES I epidemiologic follow up study. AB - Few data are available on risk for coronary heart disease in African American women with diabetes mellitus, a well-established coronary risk factor in European American women. This study tests the hypothesis that medical history of diabetes predicts coronary heart disease incidence in African American women in a national cohort. Participants in the NHANES I Epidemiologic Follow-up Study in this analysis were 1035 African American and 5732 European American women aged 25-74 years without a history of coronary heart disease. Average follow-up for survivors was 19 years (maximum 22 years). Risk of incident coronary heart disease by baseline diabetes status was estimated. Proportional hazards analyses for African American women aged 25-74 revealed significant associations of coronary heart disease risk with diabetes after adjusting for age (RR = 2.40; 95% CI, 1.58-3.64, P < 0.01). After adjusting for age, smoking, and low education, there was an elevated risk in diabetics age 25-74 (RR = 2. 34; 95% CI, 1.54-3.56, P < 0.01); this association did not differ significantly from that for European American women. Excess coronary incidence in African American compared to European American women aged 25-64 was statistically explained by controlling for diabetes history, age, education, and smoking but only partly explained by age and diabetes history. In African American women aged 25-74, diabetes was also associated with increased coronary heart disease, cardiovascular, and all-cause mortality. The population attributable risk of coronary heart disease incidence associated with a medical history of diabetes was 8.7% in African American women and 6.1% in European American women. Medical history of diabetes was a significant predictor of coronary heart disease incidence and mortality in African American women and explained some of the excess coronary incidence in younger African American compared to European American women. PMID- 10812325 TI - Showing neuropathy is related to increased mortality in diabetic patients - a survival analysis using an accelerated failure time model. AB - Diabetic subjects still have a reduced life expectancy despite many potential advances in patient care. Furthermore, population-based studies in well-defined cohorts of patients, to investigate the reduced life expectancies, are generally lacking. Computerized baseline data on a cohort of diabetic patients first attending our clinic during 1982-1985 were used to identify risk factors for increased mortality. This was carried out using an accelerated failure time (ACF) model. Out of 794 patients entered into the model, 201 (25.3%) patients died between 1982 and 1995. Baseline microvascular diabetic complications (peripheral sensory neuropathy and nephropathy) were found to be associated with increased mortality in patients, indicating that these are important, often overlooked, markers for those at greatest risk. Patients with type I (insulin dependent) diabetes mellitus were also identified as being at greater risk. PMID- 10812326 TI - Population-based study of age at menopause and ultrasound assessed carotid atherosclerosis: The Tromso Study. AB - Early menopause has been associated with higher prevalence and incidence of cardiovascular disease and death than late menopause, indicating that early loss of ovarian function and subsequent deficiency of estrogen may promote such diseases. No population-based studies have, however, examined the relation between age at menopause and atherosclerosis. We assessed the prevalence and the extent of carotid atherosclerosis by high-resolution B-mode ultrasound in 2588 postmenopausal women who participated in a population health survey. Information about age at menopause and menarche, parity, use of hormone replacement therapy, and prevalent diseases was collected, and cardiovascular risk factor levels were measured. Women with late menopause and women who ever had used postmenopausal estrogens had significantly less atherosclerosis than women with early menopause and those with never use of estrogen. This study provides further support for the hypothesis that estrogen protects women against cardiovascular disease. PMID- 10812327 TI - Performance characteristics of a two-stage dementia screen in a population sample. AB - We analyzed the performance of a two-stage screening protocol [the Modified Mini Mental State Exam (3MS) or the Informant Questionnaire for Cognitive Decline (IQCODE), and the Dementia Questionnaire (DQ)] in a weighted stratified sample of 839 subjects from a population survey of dementia in Cache County, UT. The subjects were subsequently examined using a standardized diagnostic assessment protocol. Using the method of receiver operating characteristic (ROC) analysis, the main outcome measure was area under the ROC curve (AUC). The overall AUC estimates were 0.956 (95% confidence interval 0.943-0.968) for the 3MS/IQCODE and 0.945 (0.931-0.960) for the DQ. After adjustment for age, the efficiency of the both 3MS/IQCODE and the DQ was better for subjects with genotype epsilon4/epsilon4 at APOE, better among women, and better in those with two or more years duration of dementia. The optimal threshold for this two-stage screen yielded sensitivity and specificity estimates of 91.0% and 92.0%, respectively. PMID- 10812328 TI - Community-based recruitment strategies for a longitudinal interventional study: the VECAT experience. AB - This article examines different recruitment strategies for the VECAT Study, a 4 year, double-masked, placebo-controlled, randomized clinical trial of vitamin E in the prevention of cataract and age-related maculopathy. Five recruitment methods were employed: newspaper advertising, radio advertising, approaches to community groups, approaches via general practices, and an electoral roll mail out. Participants (1204) from the community in Melbourne, Australia were recruited and enrolled within 15 months (age range: 55-80 years, mean 66 years; gender ratio: 57% female, 43% male). The electoral roll mail-out and newspaper advertising were the most efficient methods of recruitment in terms of absolute numbers of participants recruited and cost per participant. Recruitment for the VECAT study was successfully completed within the planned period. Although the electoral roll mail-out and newspaper advertising were the most efficient for this study, other methods may be of value for studies with different subject selection criteria. PMID- 10812329 TI - The pattern of sensitivity of yeast dna2 mutants to DNA damaging agents suggests a role in DSB and postreplication repair pathways. AB - The Saccharomyces cerevisiae DNA2 gene encodes a DNA-stimulated ATPase and DNA helicase/nuclease essential for DNA replication. In characterizing dna2 mutants, we have found that Dna2p also participates in DNA repair or in damage avoidance mechanisms. dna2 mutants are sensitive to X rays, although they are less sensitive than rad52 mutants. The X-ray sensitivity of dna2 mutants is suppressed by overexpression of a 5' to 3' exonuclease, the yeast FEN-1 structure-specific nuclease, encoded by the RAD27 gene, which also suppresses the growth defect of dna2-ts mutants. SGS1 encodes a helicase with similar properties to Dna2 protein. Although sgs1Delta mutants are resistant to X rays, dna2-2 sgs1Delta double mutants are more sensitive to X rays than the dna2-2 mutant. Temperature sensitive dna2 mutants are only slightly sensitive to UV light, show normal levels of spontaneous and UV induced mutagenesis, and have only a 2.5-fold elevated level of dinucleotide tract instability compared to wildtype. However, dna2Delta strains kept alive by overproduction of RAD27 are highly sensitive to UV light. These phenotypes, in addition to the epistasis analysis reported, allow us to propose that Dna2 is involved in postreplication and DSB repair pathways. PMID- 10812330 TI - Efficient repair of hydrogen peroxide-induced DNA damage by Escherichia coli requires SOS induction of RecA and RuvA proteins. AB - The survival of Escherichia coli following treatment with a low dose (1-3 mM) of hydrogen peroxide (H(2)O(2)) that causes extensive mode-one killing of DNA repair mutants is stimulated by the induction of the SOS regulon. Results for various mutants indicate that induction of recA and RecA protein-mediated recombination are critical factors contributing to the repair of H(2)O(2)-induced oxidative DNA damage. However, because DNA damage activates RecA protein's coprotease activity essential to cleavage of LexA repressor protein and derepression of all SOS genes, it is unclear to what extent induction of RecA protein stimulates this repair. To make this determination, we examined mode-one killing of DeltarecA cells carrying plasmid-borne recA (P(tac)-recA(+)) and constitutively expressing a fully induced level of wild-type RecA protein when SOS genes other than recA are non-inducible in a lexA3 (Ind(-)) genetic background or inducible in a lexA(+) background. At a H(2)O(2) dose resulting in maximal killing, DeltarecA lexA3 (Ind(-)) cells with P(tac)-recA(+) show 40-fold greater survival than lexA3 (Ind(-)) cells with chromosomal recA having a low, non-induced level of RecA protein. However, they still show 10- to 15-fold lower survival than wild-type cells and DeltarecA lexA(+) cells with P(tac)-recA(+). To determine if the inducible RuvA protein stimulates survival, we examined a ruvA60 mutant that is defective for the repair of UV-induced DNA damage. This mutant also shows 10- to 15-fold lower survival than wild-type cells. We conclude that while induction of RecA protein has a pronounced stimulatory effect on the recombinational repair of H(2)O(2)-induced oxidative DNA damage, the induction of other SOS proteins such as RuvA is essential for wild-type repair. PMID- 10812331 TI - Effect of age on the formation and repair of UV photoproducts in human skin in situ. AB - Ultraviolet radiation (UVR)-induced photoproducts can be measured by a number of methods. The newly developed 32P-postlabelling method is feasible in molecular epidemiological studies due to its sensitivity, specificity and little amount DNA needed. We applied the 32P-postlabelling method to investigate the induction and repair of photoproducts (cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts) after UVR in human skin in situ and studied the effects of age, skin type and gender. The study included 30 subjects aged 32-78 years. The photoproduct induction levels varied 7- to 15-fold between the individuals tested. All four types of photoproducts were induced at a higher frequency in the older population (>/=50 years) than in the younger population (<50 years). Individuals with skin type I and II had a higher CPD induction frequency than individuals with skin type III and IV. In both cases, the differences in thymidylyl (3'-5') thymidylyl (3'-5')-2'-deoxycytidine induction reached statistical significant levels (p<0.05). Photoproduct repair rates 24 h and 48 h after UV irradiation showed a large inter-individual variation. No clear effects of age, skin type or gender on DNA repair could be detected. Our data suggest that UV-induced DNA photoproduct levels increase with age. PMID- 10812332 TI - Elevation of sister chromatid exchange in Saccharomyces cerevisiae sgs1 disruptants and the relevance of the disruptants as a system to evaluate mutations in Bloom's syndrome gene. AB - The SGS1 of Saccharomyces cerevisiae is a homologue of the Bloom's syndrome and Werner's syndrome genes. The sgs1 disruptants show hyperrecombination, higher sensitivity to methyl methanesulfonate and hydroxyurea, and poor sporulation. In this study, we found that sister chromatid exchange was increased in sgs1 disruptants. We made mutated SGS1 genes coding a protein proved to lack DNA helicase activity (sgs1-hd), having equivalent missense mutations found in Bloom's syndrome patients (sgs1-BS1, sgs1-BS2). None of the mutated genes could suppress the higher sensitivity to methyl methanesulfonate and hydroxyurea and the increased frequency of interchromosomal recombination and sister chromatid exchange of sgs1 disruptants. On the other hand, all of the mutant genes were able to complement the poor sporulation phenotype of sgs1 disruptants, although the values were not as high as that of wild-type SGS1. PMID- 10812333 TI - Effects of epigallocatechin gallate and quercetin on oxidative damage to cellular DNA. AB - Phenolic phytochemicals are thought to promote optimal health, partly via their antioxidant effects in protecting cellular components against free radicals. The aims of this study were to assess the free radical-scavenging activities of several common phenolic phytochemicals, and then, the effects of the most potent phenolic phytochemicals on oxidative damage to DNA in cultured cells. Epigallocatechin gallate (EGCG) scavenged the stable free radical, alpha,alpha diphenyl-beta-picrylhydrazyl (DPPH), most effectively, while quercetin was about half as effective. Genistein, daidzein, hesperetin, and naringenin did not scavenge DPPH appreciably. Jurkat T-lymphocytes that were pre-incubated with relatively low concentrations of either EGCG or quercetin were less susceptible to DNA damage induced by either a reactive oxygen species or a reactive nitrogen species, as evaluated by the comet assay. More specifically, control cells had a comet score of only 17+/-5, indicating minimal DNA damage. Cells challenged with 25 microM hydrogen peroxide (H(2)O(2)) or 100 microM 3-morpholinosydnonimine (SIN 1, a peroxynitrite generator) had comet scores of 188+/-6 and 125+/-12, respectively, indicating extensive DNA damage. The H(2)O(2)-induced DNA damage was inhibited with 10 microM of either EGCG (comet score: 113+/-23) or quercetin (comet score: 82+/-7). Similarly, the SIN-1-mediated DNA damage was inhibited with 10 microM of either EGCG (comet score: 79+/-13) or quercetin (comet score: 72+/-17). In contrast, noticeable DNA damage was induced in Jurkat T-lymphocytes by incubating with 10-fold higher concentrations (i.e., 100 microM) of either EGCG (comet score: 56+/-17) or quercetin (comet score: 64+/-13) by themselves. Collectively, these data suggest that low concentrations of EGCG and quercetin scavenged free radicals, thereby inhibiting oxidative damage to cellular DNA. But, high concentrations of either EGCG or quercetin alone induced cellular DNA damage. PMID- 10812334 TI - Nucleotide excision repair endonuclease genes in Drosophila melanogaster. AB - Nucleotide excision repair (NER) is the primary pathway for the removal of ultraviolet light-induced damage and bulky adducts from DNA in eukaryotes. During NER, the helix is unwound around the damaged site, and incisions are made on the 5' and 3' sides, to release an oligonucleotide carrying the lesion. Repair synthesis can then proceed, using the intact strand as a template. The incisions flanking the lesion are catalyzed by different structure-specific endonucleases. The 5' incision is made by a heterodimer of XPF and ERCC1 (Rad1p-Rad10p in Saccharomyces cerevisiae), and the 3' incision is made by XPG (Rad2p in S. cerevisiae). We previously showed that the Drosophila XPF homologue is encoded by the meiotic recombination gene mei-9. We report here the identification of the genes encoding the XPG and ERCC1 homologues (XPG(Dm) and ERCC1(Dm)). XPG(Dm) is encoded by the mus201 gene; we found frameshift mutations predicted to produce truncated XPG(Dm) proteins in each of two mus201 alleles. These mutations cause defects in nucleotide excision repair and hypersensitivity to alkylating agents and ultraviolet light, but do not cause hypersensitivity to ionizing radiation and do not impair viability or fertility. ERCC1(Dm) interacts strongly in a yeast two-hybrid assay with MEI-9, indicative of the presumed requirement for these polypeptides to dimerize to form the functional endonuclease. The Drosophila Ercc1 gene maps to polytene region 51D1-2. The nucleotide excision repair gene mus210 maps nearby (51E-F) but is distinct from Ercc1. PMID- 10812335 TI - Mutagenic effects of gamma-rays and incorporated 8-3H-purines on extracellular lambda phage: influence of mutY and mutM host mutations. AB - The lethal and mutagenic effects on phage lambdacI857 of 60Co gamma-rays and of decay of 3H incorporated into phage DNA both as 8-3H-deoxyadenosine and 8-3H deoxyguanosine (using 8-3H-adenine as a labelled DNA precursor) were studied on four isogenic Escherichia coli strains: AB1157 M(+)Y(+) (wild type, mutM(+) mutY(+)), AB1157 M(-)Y(+) (mutM::kan mutY(+) mutant deficient in the formamidopyrimidine-DNA glycosylase MutM), AB1157 M(+)Y(-) (mutM(+) mutY mutant deficient in the A:G mismatch DNA glycosylase MutY), and AB1157 M(-)Y(-) (mutM::kan mutY double mutant deficient in both DNA glycosylases). The main products of transmutation component of 3H decay in position 8 of purine residues are 8-oxo-7, 8-dihydroadenine (8-oxoA) and 8-oxo-7,8-dihydroguanine (8-oxoG), the latter being responsible for the most part of the mutagenic effect. The lethal effects of both gamma-rays and tritium decay virtually did not depend on the repair phenotypes of the host strains used. Therefore, the MutM and MutY glycosylases are not involved in the repair of lethal DNA damages induced by ionizing radiation or by the transmutation component of 3H decay in purine residues of phage DNA. The efficiencies of mutagenic action of 3H-purines E(m) (frequencies of c-mutations per one 3H decay in phage genome) were 2.4-, 3.8- and 55-fold higher in the M(-)Y(+), M(+)Y(-) and M(-)Y(-) mutants, respectively, in comparison to the wild-type host. The mutagenic efficiencies E(m) for gamma-rays were nearly identical in the M(+)Y(+) and M(-)Y(+) hosts, but were increased 1.8- and 8.3-fold, respectively, in the M(+)Y(-) and M(-)Y(-) mutants. These data suggest that: (1) the MutY and MutM DNA glycosylases are important for prevention of mutations caused not only by spontaneous oxidation of guanine residues, but also by ionizing radiation or by decay of 3H incorporated into purine bases of DNA; (2) the MutY and MutM enzymes functionally cooperate in elimination of mutagenic damages induced by these agents. PMID- 10812337 TI - Institute of Medicine report. PMID- 10812336 TI - Autonomous 3'-->5' exonucleases can proofread for DNA polymerase beta from rat liver. AB - Autonomous 3'-->5'exonucleases are not bound covalently to DNA polymerases but are often involved in replicative complexes. Such exonucleases from rat liver, calf thymus and Escherichia coli (molecular masses of 28+/-2 kDa) are shown to increase more than 10-fold the accuracy of DNA polymerase beta (the most inaccurate mammalian polymerase) from rat liver in the course of reduplication of the primed DNA of bacteriophage phiX174 amber 3 in vitro. The extent of correction increases together with the rise in 3'-->5' exonuclease concentration. Extrapolation of the in vitro DNA replication fidelity to the cellular levels of rat exonuclease and beta-polymerase suggests that exonucleolytic proofreading could augment the accuracy of DNA synthesis by two orders of magnitude. These results are not explained by exonucleolytic degradation of the primers ("no synthesis-no errors"), since similar data are obtained with the use of the primers 15 or 150 nucleotides long in the course of a fidelity assay of DNA polymerases, both alpha and beta, in the presence of various concentrations of 3' ->5' exonuclease. PMID- 10812338 TI - Reporting complications on a general surgery service. PMID- 10812339 TI - Soft-tissue images. Bilateral large iliac artery aneurysms. PMID- 10812340 TI - Musculoskeletal images. Secondary signs of patellofemoral dislocation. PMID- 10812341 TI - Soft-tissue case 32. Right colonic intramural hematoma. PMID- 10812342 TI - Musculoskeletal case 9. Elastofibroma dorsi. PMID- 10812343 TI - Is Canadian cardiac surgeons' management of asymptomatic carotid artery stenosis at coronary artery bypass supported by the literature? A survey and a critical appraisal of the literature. AB - OBJECTIVE: To document and compare the management of asymptomatic carotid stenosis (ACS) by Canadian cardiac surgeons at coronary artery bypass grafting (CABG) against a critical literature analysis. DESIGN: A multiple choice survey and a structured literature review. DATA SOURCES: Seventy-seven surgeons and 272 publications selected from the English literature between 1980 and 1997. Search terms used were "carotid," "coronary bypass," and "cardiac surgery." STUDY SELECTION: Five natural history studies were identified, and 58 studies were found that had objective documentation of ACS of 50% or more before cardiac surgery, and both operative stroke and mortality data reported for CABG with and without carotid endarterectomy (CEA). DATA EXTRACTION: Natural history and outcome studies were independently rated against published guidelines. Outcome data were independently pooled and compared. Data discrepancy was resolved by consensus. Survey results were tabulated for simple descriptive statistics. DATA SYNTHESIS: No methodologically sound natural history studies were found to document an increased risk of stroke from ACS after CABG. There were no randomized controlled studies to guide treatment recommendations. Pooled data for stroke or death did not support CEA for risk reduction from ACS at CABG (relative risk 0.9, p = 0.5). Ninety-four percent of surgeons believed that the literature is insufficient to support the routine use of CEA to reduce the risk of stroke from ACS after CABG. Despite this, 20% of surgeons advocated CEA for this purpose. CONCLUSION: The management of ACS at CABG by the majority of Canadian cardiac surgeons is consistent with the results of the literature review; however, significant management variation exists. PMID- 10812344 TI - Assessment of patient waiting times for vascular surgery. AB - OBJECTIVES: To assess patient waiting times for vascular surgery and to determine if complications of the disease develop while the patients are awaiting surgery. DESIGN: Prospective cohort study. SETTING: A university-affiliated tertiary care institution. PATIENTS: All 554 patients who underwent scheduled vascular surgical procedures between April 1995 and October 1996. OUTCOME MEASURES: A literature review carried out to develop guidelines for acceptable waiting times for surgery associated with various vascular disorders based on their natural history (benchmark target); actual waiting times, defined as the interval from the date each patient was booked for surgery to the date of admission to hospital for the procedure; the proportion of patients admitted within the benchmark targets; and whether prolonged waiting time placed patients at risk for complications of their disease. RESULTS: Of the 554 patients, 382 (69%) were admitted within the benchmark waiting times. Of 84 patients having an abdominal aortic aneurysm, the aneurysm ruptured during the waiting period in 6, and 4 of them died, for a complication rate of 7% and a death rate of 5%. Two of the 6 aneurysms ruptured after the patient had waited longer than the target time. Three of 100 patients with symptomatic carotid artery stenosis awaiting admission for carotid endarterectomy suffered ischemic stroke, for a 3% complication rate; all had waited longer than the target period. One patient suffered occlusion of a femoropopliteal bypass graft while awaiting revision of a stenosed bypass graft. CONCLUSIONS: This study suggests that although most patients are admitted for operation within the benchmark time, one-third are admitted late and may suffer serious complications of their disease while awaiting admission for the procedure. PMID- 10812345 TI - Complications on a general surgery service: incidence and reporting. AB - OBJECTIVES: To determine the incidence and nature of complications on a general surgery service and to compare these results with pre-existing institutional recording and reporting methods. DESIGN: A single observer prospectively monitored the presence and documentation of complications for all patients admitted to the general surgery service at the Wellesley Central Hospital over a 2-month period, through daily chart reviews, attendance at rounds and surgical operating rooms, frequent patient visits on the ward and interviews with the health care team. SETTING: The general surgery service of an urban, university affiliated teaching hospital. PATIENTS: One hundred and ninety-two general surgery inpatients over 1277 patient-days from June 16, 1996, until Aug. 15, 1996. Same-day surgery patients were not included. RESULTS: Seventy-five (39%) of the 192 patients suffered a total of 144 complications. Two complications (1%) were fatal, 10 (7%) were life threatening, 90 (63%) were of moderate severity and 42 (29%) were trivial. Of these 144 complications, 26 (18%) were deemed potentially attributable to error. One hundred and twelve (78%) of the complications occurred during or after a surgical operation and were related directly or indirectly to it. Only 9 (6%) complications were not documented in the progress notes of the patients' charts. However, 115 (80%) were not presented at weekly morbidity and mortality rounds, and 95 (66%) were not documented on the face sheet of the patients' final medical records. CONCLUSIONS: Complications are common and are underreported by traditional methods. Since hospital funding and quality improvement efforts depend on accurate identification and recording of adverse events, strategies to improve the recording and reporting of complications must be developed. PMID- 10812346 TI - The treatment of pseudoarthrosis of the scaphoid by bone grafting and three methods of internal fixation. AB - OBJECTIVES: To measure the rate of union in patients with pseudoarthosis of the scaphoid, treated with trapezoidal bone grafting as outlined by Fernandez and 1 of 3 methods of internal fixation and to compare unions versus nonunions and potential predictors of union to determine if associations exist. DESIGN: A retrospective radiologic study of scaphoid pseudoarthroses. SETTING: Division of Orthopedic Surgery, Ottawa Hospital, General Site, a tertiary care facility. PATIENTS: Thirty-four patients with nonunion of scaphoid fractures, treated between 1990 and 1997, with an average follow-up of 19.8 months. INTERVENTIONS: Trapezoidal bone grafting and internal fixation with Kirschner (K) wires, an AO cannulated screw or a Herbert screw. OUTCOME MEASURES: The time to union of scaphoid pseudoarthroses and predictors of union, including the classification, location of pseudoarthrosis, type of internal fixation and length of bone graft. RESULTS: The results showed a correlation between the classification and location of the fracture as determined radiologically, and the outcome. There was no correlation between the type of internal fixation used and the outcome, or between the length of the bone graft and the outcome. Twenty-three patients had radiologically demonstrated union after a mean time of 8.2 months; 16 of 24 patients achieved successful union when treated with K-wire implants, after a mean time of 7.2 months. CONCLUSIONS: Trapezoidal bone grafting and internal fixation with K wires is a practical technique, classification and location of the fracture notwithstanding. Time to union is long, and the results may be unpredictable. Use of K wires for internal fixation presents the clinician with an alternative to fixation with either the AO cannulated screw or the Herbert screw, and has the advantages of cost, ease of insertion and accessibility. This method may therefore be the treatment of choice in developing countries. Resection of the area of pseudoarthrosis must include all fibrous tissue and sclerotic bone. The length of graft, within the parameters of this study, did not affect the outcome. PMID- 10812347 TI - An analysis of a preoperative pediatric autologous blood donation program. AB - OBJECTIVE: To determine the efficacy of a pediatric autologous blood donation program. DESIGN: A retrospective study of patient charts and blood-bank records. SETTING: The Children's Hospital of Eastern Ontario, Ottawa, a tertiary care, pediatric centre. PATIENTS: One hundred and seventy-three children who received blood transfusions for a total of 182 procedures between June 1987 and June 1997. INTERVENTIONS: Autologous and homologous blood transfusion required for major surgical intervention, primarily spinal fusion. MAIN OUTCOME MEASURES: Surgeons' accuracy in predicting the number of autologous blood units required for a given procedure, compliance rate (children's ability to donate the requested volume of blood), utilization rate of autologous units and rate of allogeneic transfusion. RESULTS: The surgeons' accuracy in predicting the number of autologous units required for a given procedure was 53.8%. The compliance rate of children to donate the requested amount of blood was 80.3%. In children below the standard age and weight criteria for blood donation the compliance rate was 75.5%. The utilization rate of autologous units obtained was 84.4% and the incidence of allogeneic transfusion was 26.6%. CONCLUSIONS: There was a high rate of compliance and utilization of predonated autologous blood in the children in the study. Preoperative blood donation programs are safe and effective in children, even in those below the standard age and weight criteria of 10 years and 40 kg. PMID- 10812348 TI - Autopsies and death certification in deaths due to blunt trauma: what are we missing? AB - OBJECTIVES: To determine the frequency, body region and severity of injuries missed by the clinical team in patients who die of blunt trauma, and to examine the accuracy of the cause of death as recorded on death certificates. DESIGN: A retrospective review. SETTING: London Health Sciences Centre, London, Ont. PATIENTS: One hundred and eight deaths due to blunt trauma occurring during the period Apr. 1, 1991, to Mar. 31, 1997. Two groups were considered: clinically significant missed injuries were identified by comparing patient charts only (group 1) and more detailed injury lists from the autopsies and charts of the patients (group 2). OUTCOME MEASURES: Chart and autopsy findings. RESULTS: Of the 108 patients, 78 (72%) were male, and they had a median age of 39 years (range from 2 to 90 years). The most common cause of death was neurologic injury (27%), followed by sepsis (17%) and hemorrhage (15%). There was disagreement between the treating physicians and the causes of death listed on the death certificate in 40% of cases and with the coroner in 7% of cases. Seventy-seven clinically significant injuries were missed in 51 (47%) of the 108 patient deaths. Injuries were missed in 29% of inhospital deaths and 100% of emergency department deaths. Abdominal and head injuries accounted for 43% and 34% of the missed injuries, respectively. CONCLUSIONS: The information contained on the death certificate can be misleading. Health care planners utilizing this data may draw inaccurate conclusions regarding causes of death, which may have an impact on trauma system development. Missed injuries continue to be a concern in the management of patients with major blunt trauma. PMID- 10812349 TI - Intraosseous paraganglioma of the sacrum: a case report. PMID- 10812350 TI - Strut allograft invasion by Paget's disease of bone: a case report. PMID- 10812351 TI - Simple exposure of the internal thoracic artery for minimally invasive coronary artery bypass. AB - A simple method is described for harvesting a pedicle of internal thoracic artery for minimally invasive coronary artery bypass grafting with the use of a small scapular retractor. The technique has been used successfully in 39 patients. All patients were followed up early postoperatively by angiography, and the patency rate was 96%. Follow-up angiography (mean [and standard error] 9.6 [4.8] months) was carried out in 63% of the patients, demonstrating a cumulative patency rate of 95.4% with no evidence of steal. None of the patients died and there were no cases of myocardial infarction perioperatively. Recurrent angina developed in 2 patients. This technique is safe, effective and inexpensive. PMID- 10812352 TI - Percutaneous fine-needle aspiration biopsy for lung masses. PMID- 10812353 TI - Ischemic Meckel's diverticulum and acute appendicitis. PMID- 10812354 TI - [Cancer of the uterine cervix]. PMID- 10812355 TI - [Imaging of cancer of the uterine cervix]. AB - Imaging of uterine cervix carcinoma has evolved during the last decade. Recent developments in magnetic resonance imaging have expanded the role of MRI in evaluating the pathology of uterine cervix carcinoma. MRI is now the modality of choice for tumor staging, evaluating tumor response to treatment, diagnosing recurrences and for evaluating pregnant patients. MRI images will soon be used to calculate dosimetry for brachytherapy with matching and fusion software. PMID- 10812357 TI - [New approaches in "conventional" surgery of uterine cervix cancer]. AB - The radical hysterectomy by laparotomy with resection of the proximal and distal parts of the parametria (type III according to the Piver's classification) is the most appropriate treatment of stage Ib and II cervical carcinoma. Nevertheless, a resection of a partial part of the parametria and conservative surgery (of the uterus and/or ovaries) are possible in selected cases of patients with good prognostic factors. The lymphadenectomy has a diagnostic, prognostic but probably also a therapeutic value. Para-aortic lymphadenectomy should be performed in patients with a bulky tumor. Pelvic exenteration with reconstructive surgery could be discussed in young patients with centro-pelvic recurrence of a cervical carcinoma. PMID- 10812356 TI - [Role of laparoscopy in the management of uterine cervix cancer]. AB - At the turn of this century, the evidence of the benefits of a concurrent chemo radiotherapy in locally advanced tumors and the development of mini-invasive surgery (laparoscopic and radical vaginal surgery) are the two main advances in the management of cervical carcinomas. From a personal experience of 304 cervical carcinomas, the different techniques of laparoscopy used in cervical carcinomas are addressed and discussed. Their long-term results when involved in the management protocols of cervical carcinomas at different stages are reported. From this series, some conclusions are drawn: 1) laparoscopy can spare a laparotomy in early-stage node-negative patients with low tumoral volume; 2) it can spare a systematic extended-field radiation therapy in high-risk patients with node-negative para-aortic exploration; 3) it can spare surgery in patients with a centro-pelvic advanced stage or recurrence, possibly candidates for an exenterative procedure, if occult spread is found in the intra- or retroperitoneal areas. The more and more frequent combination of the mini invasive surgery for staging and treatment and radiotherapy or chemotherapy explains the need for new protocols of a more and more complex and specialized management. PMID- 10812358 TI - [Contribution of imaging in radiotherapy planning in uterine cervix cancers]. AB - External irradiation and brachytherapy are curative in the treatment of carcinoma of the cervix. The aim of radiotherapy is to optimize the irradiation of the target volume and to reduce the dose to critical organs. The use of imaging (computed tomography and magnetic resonance imaging added to clinical findings and standard guidelines) are studied in the treatment planning of external irradiation and brachytherapy in carcinoma of the cervix. Imaging allows an individualized and conformal treatment planning. PMID- 10812359 TI - [Curietherapy in uterine cervix cancers: what therapeutic trends?]. AB - Brachytherapy is a fundamental step in the treatment of patients with cervical cancer. Brachytherapy allows a significant increase in the local control rate as well as the survival rate. Brachytherapy has to be performed as soon as possible after external irradiation in order to maintain the overall treatment time below 53 days. Technical and dosimetric data characterizing low dose-rate brachytherapy using ICRU 38 recommendations have led to an improvement in local control and a decrease in complications. Data are less well known for other dose rates. The role of interstitial brachytherapy is not clearly defined and its potential benefit is probably balanced by an increase in severe complications. Concomitant brachy-chemotherapy requires further clinical investigations, even if concomitant radio-chemotherapy has become a standard in advanced cervical cancers. PMID- 10812360 TI - [Radiochemotherapy of uterine cervix cancers. Recent data]. AB - Low-stage uterine cervix carcinoma can be treated by either surgery, radiation therapy or combined treatments with high cure rates ranging from 90 to 95% for stage IB1 tumors. However, the standard treatment, combining external beam plus intracavitary radiation, fails to control the progression of the disease in 35 to 90% of patients with locally advanced cervical cancer. No substantial improvements have been made in the treatment of these tumors in the past two decades. The addition of concurrent 5-FU in a phase III study failed to improve the results in the overall patient population, but the five-year DFS was significantly better in a subset of patients (tumor > 5 cm and IB/IIA or medial parametrial IIB disease). Concurrent chemoradiation and adjuvant chemotherapy with epirubicin showed, in a phase III study, a significantly longer DFS in patients treated with chemotherapy despite the same long-term local tumor control. After many phase II studies, five phase III studies have recently demonstrated a 40 to 60% reduction in the relative risk of recurrence with cisplatin-containing chemoradiation. Across these studies, the risk of death was reduced by 30 to 50%. The benefit was less clear in patients with stages III-IV tumors than in patients with lower stages associated with poor prognostic factors. Haematologic and gastrointestinal toxicity of chemoradiation was greater than that of radiotherapy alone. However, late side effects were similar in the different treatment groups. These results must be confirmed with a longer follow up. The importance of concurrent chemotherapy during the brachytherapy procedure should be analyzed. It has yet to be determined which chemotherapy regimen achieves the most favorable therapeutic ratio. PMID- 10812361 TI - [Complications of treatments of invasive cancers of the uterine cervix with intact uterus: results and prevention]. AB - The prospective record of acute and late toxicity after treatment of cervix carcinomas is a part of the description of treatment outcome as well as local control and survival. Due to the large number of scales and glossaries used, the comparison of the results from one study to another is often difficult. The French-Italian syllabus seems to be the most reliable scale, providing implementation of quality of life assessments. The main predictive factor of complications, which is not related to the treatment type, is the previous history of abdominal or pelvic surgery. The incidence and severity of complications occurring after surgery are related to the surgical procedure and to the amount of peri-uterine tissues removed. The increase in dose and volume of external irradiation and brachytherapy and the increase in dose rate of the low dose rate brachytherapy are responsible for the radiotherapeutic morbidity. The significant decrease of severe complication rates during the last 15 years was obtained by the implementation of individual adjustments in treatment planning. The treatment strategies of early bulky and advanced carcinomas are changing. Concurrent radiotherapy and chemotherapy is becoming a standard, but its late toxicity needs to be documented by a longer follow-up. The optimisation of radiation therapy should remain a reference to evaluate the outcome and morbidity of the new combined strategies: the addition of chemotherapy will never compensate for less than optimal radiotherapy/brachytherapy planning. PMID- 10812363 TI - [Treatment of stroke in Germany]. PMID- 10812362 TI - Definitive radiotherapy based on HDR brachytherapy with iridium 192 in uterine cervix carcinoma: report on the Vienna University Hospital findings (1993-1997) compared to the preceding period in the context of ICRU 38 recommendations. AB - According to the reports described in the literature, fractionated HDR brachytherapy seems to represent one option for the primary treatment of cervical carcinoma. In order to render such treatment transparent and comparable for those interested in the field, we have attempted to report our recent experience obtained in Vienna from 1993-1997 using the terminology proposed by the ICRU report 38, focusing in particular on dose and volume reporting and a linear quadratic model. Based on these parameters, a comparison with the preceding period in Vienna (LDR/HDR) has been made, with an attempt to correlate different methods and parameters with outcome. MATERIAL AND METHODS: One hundred and eighty nine patients (mean age 67 years) were treated with curative intent (stage Ia: 2, Ib: 11, IIa: 11, IIb: 79, IIIa: 19, IIIb: 59, IVa: 5, IVb: 3 patients) using a combination of intracervical high-dose rate (HDR) brachytherapy (ring-tandem applicator) and a box technique for external-beam therapy (EBT: 48.6-50 Gy, linac 25 MV). Small tumors were treated with 5-6 fractions of 7 Gy at point A and 50 Gy EBT (25 Gy in the brachytherapy reference volume) which is isoeffective to 76-86 Gy at point A. Large tumors received 3-4 fractions of 7 Gy after 50 Gy EBT with open fields, which is isoeffective to 82-92 Gy at point A. TRAK varied from mean 1.4 cGy (3 fractions) to 2.8 cGy (6 fractions) at one meter. 3-D treatment planning for brachytherapy was based on conventional X-rays and in 181/189 patients on computed tomography (CT) with the applicator in place. Computer calculated volumes of the brachytherapy reference isodose (7 Gy/fraction) ranged from 46-155 ccm (mean 87 ccm); the respective mean hwt-volume (height x width x thickness) was 180 ccm. The 60 Gy HWT volumes (25 Gy from EBT) for the irradiation of small tumors ranged from 240 to 407 ccm (mean 337 ccm) and for larger tumors (50 Gy for EBT) from 452 to 785 ccm (mean 607 ccm). The mean dose for brachytherapy was 16.2 Gy (6.2-37.8 Gy) at the ICRU rectum reference point and 14.4 Gy (4.6-35.7 Gy) at the ICRU bladder point. Taking into account the dose for EBT, the mean isoeffective dose at the ICRU rectum reference point was 69.9 Gy (28.4-98.7 Gy). Overall treatment time was six weeks for small tumors and eight weeks for large tumors. RESULTS: After a mean follow-up of 34 months, actuarial pelvic control and disease-specific survival rates at three years were 77.6/68.6% for all patients, 100/77.1% for stage Ib, 100/100% stage IIa, 87.0/78.0% stage IIb, 52.7/52.1% stage IIIa, 69.1/58.6% stage IIIb and 60/53.3% stage IVa. According to the LENT/SOMA score which had been prospectively introduced, the actuarial late complication rate for grades 3 and 4 was 2.9% for the bladder, 4.0% for the bowel, 6.1% for the rectum and 30.6% for the vagina (shortening and obliteration); in total for all grades 23.6, 18.4, 24.2, and 67.6%, respectively. CONCLUSION: In our experience, HDR brachytherapy combined with EBT is an efficient method if sufficient radiation doses and volumes are applied, both with regard to tumor control and adverse side effects. In future, the therapeutic window will be increased by systematic integration of magnetic resonance imaging (MRI) into treatment planning, thus allowing for a highly individualized approach with further adaptation of radiation dose and volume both to the target and to the individual topography of organs at risk. PMID- 10812364 TI - [Stroke unit at a medical clinic providing general medical care. Model and initial clinical results]. AB - BACKGROUND AND OBJECTIVES: To achieve in Germany complete therapeutic cover for patients after a stroke, models will have to be formulated that incorporate such units within the confines of departments of general medicine, where most stroke patients are at present being treated. Preliminary experience of such a unit is reported here. PATIENTS AND METHODS: The stroke unit, conceived for "mixed assessment", has 6 beds within a normal ward but an increased number of specially trained personnel and specialized equipment. The aim was the rapid diagnosis, targeted treatment, monitoring, early mobilization and, if necessary, smooth and direct transfer to an installation for rehabilitation to which the patient can be taken during the entire stay in the acute stroke unit. RESULTS: During a period of 11 months 120 patients (68 men, 52 women; mean age 71 years) were treated in the stroke unit, 47% having been admitted within 3 hours of the acute episode. An acute ischaemic attack had occurred in 85%, cerebral haemorrhage in 15%. The median European stroke scale (ESS) was 76 on admission, 90.5 on discharge, the Barthel index 45 and 90, the ranking scale 3.2 and 2.3, respectively. The complication rate per patient during hospital stay was 0.5. There were 12 deaths (10%), 22.2% among those with cerebral bleeding, 7.7% of those with ischaemia. Mean hospital stay was 13.3 days. 57.4% patients were transferred to a rehabilitation unit, 34.2% discharged home, 2.8% directly to a nursing home. The remaining 6 had undergone immediate vascular (4) or neurological (2) procedures. CONCLUSION: A "mixed assessment" stroke unit within a general medical department is feasible. It improves working conditions and patient care as well as reducing the length of hospital stay. PMID- 10812365 TI - [Knowledge about stroke among the German population]. AB - BACKGROUND AND OBJECTIVE: Modern stroke therapy requires patients to correctly identify stroke symptoms and seek immediate hospital admission. US studies showed that only 57% of the population knew at least one stroke symptom. This is the first study about stroke knowledge among German populations. METHODS: Using a cross-sectional questionnaire survey, 300 working-age participants of the PROCAM study, the Prospective Cardiovascular Munster Study, and 95 senior citizens of the Augsburg Study, a follow-up project of the MONICA survey 1989/90, were asked about stroke symptoms and what to do if they occur. Good knowledge about stroke was defined as knowing at least two stroke symptoms and calling the emergency medical system or seeking immediate hospital admission in case of symptoms. RESULTS: Participation rate in the PROCAM study was 90%, while all senior citizens took part. The mean age of the working population was 41.2 years, the mean age of the retired population was 72.8 years. 35% of the working and 24.5% of the retired participants knew at least two stroke symptoms. Urgent hospital admission was selected by 78.2% of the occupational but only 41.5% of the retired participants. Good stroke knowledge was demonstrated by nearly a third of the workers but less than 10% of the elderly. Among the occupational population, being a white-collar worker or knowing someone with a stroke was a significant predictor of good stroke knowledge. Among senior citizens higher age and current smoking status were significant predictors. CONCLUSION: Our study shows significant information deficits about stroke in our population: education needs to be geared especially towards the elderly. PMID- 10812366 TI - [Multilocular thyroid gland ectopy]. AB - HISTORY AND CLINICAL FINDINGS: A 86-year-old woman was hospitalized two days before death. Her past history included essential hypertension and joint pain. Electrocardiography and laboratory findings revealed an acute myocardial infarction. Chemical laboratory examination demonstrated hyperthyroidism. The x ray of the chest showed a tumor-like mass in the right lung and a nodular goiter with focal changes. Sonographically a tumor in the left colon was diagnosed. Inspite of intensive care the patient died two days later of cardiogenic shock. AUTOPTIC DIAGNOSIS: The autopsy revealed a transmural myocardial infarction with rupture of the heart wall. An adenocarcinoma of the rectum infiltrating the perirectal fatty tissue was diagnosed. Metastases were absent. Additional to an eutopic nodular goiter there was ectopic thyroid tissue in the lung, as a tumour mass under the visceral pleura, in the pelvic cavity and in the skeleton. The histologic findings revealed a close resemblance to the thyroid gland in normal anatomical position. In small foci in the eutopic and ectopic thyroid tissues there were signs of hyperthyroidism. There was no evidence of malignancy. CONCLUSION: One should always keep in mind that manifest hyperthyroidism, not explicable on the grounds of the thyroid findings in normal anatomical position can point to ectopic (multilocular) thyroid tissue, especially when there are "tumours" of uncertain origin. PMID- 10812367 TI - [Hypovolemic shock associated with generalized edema: paroxysmal non-hereditary angioedema (Clarkson syndrome)]. PMID- 10812368 TI - [Airway stents. A skeptical plea for palliative medicine]. PMID- 10812369 TI - ["Meanwhile to disdain medicine is not foolishness but an outrage": Melanchthon (1497-1560) and the art of medicine]. PMID- 10812370 TI - [Violation of traffic regulations by physicians on emergency calls. Decision of the Bavarian High Regional Court on November 22, 1999]. PMID- 10812371 TI - Foodservice in correctional facilities. PMID- 10812372 TI - The antioxidant conundrum: two recent studies point in different directions. PMID- 10812373 TI - Diet office redesign to enhance satisfaction and reduce costs. PMID- 10812374 TI - Proposed dietary guidelines 2000 generate comments, controversy. PMID- 10812375 TI - Dietetics: legalities, ethics, and eccentricities. PMID- 10812376 TI - Improved lactose digestion and intolerance among African-American adolescent girls fed a dairy-rich diet. AB - OBJECTIVE: To determine whether African-American adolescent girls who were fed a dairy-rich diet for 21 days could adapt to lactose, experiencing an overall improvement in lactose tolerance as well as a decrease in hydrogen gas production. DESIGN: Twenty-one-day dietary intervention study. SUBJECTS/SETTING: Seventeen of 21 African-American girls (aged 11 to 15 years) enrolled in a calcium metabolism study chose to participate in the lactose tolerance study. Subjects were screened for any diseases, conditions, or medications that might alter calcium metabolism or colonic fermentation. Subjects were housed in a fraternity on the Purdue University, West Lafayette, Ind, campus, and were supervised 24 hours a day. INTERVENTION: Subjects consumed a dairy-based diet averaging 1,200 mg calcium and 33 g lactose per day for 21 days. Lactose digestion was assessed by an 8-hour breath hydrogen test on days 1 and 21, and symptoms of intolerance (abdominal pain, bloating, flatulence, and diarrhea) were evaluated hourly on a ranked scale during the breath hydrogen tests and once each evening during the 21-day feeding period. MAIN OUTCOME MEASURES: A comparison of breath hydrogen production and gastrointestinal symptoms at the beginning and end of the study. STATISTICAL ANALYSES PERFORMED: The Wilcoxon signed ranks test was used to compare the area under the curve for the 2 breath hydrogen tests. Spearman's p test for trend was used to determine whether there was a change in symptoms. All statistical analyses were 2-tailed and significance was set at P = .05. RESULTS: Fourteen of the 17 subjects had lactose maldigestion. Breath hydrogen excretion decreased significantly (P < .03) from the beginning (148.3 +/ 27.0 ppm x hours) to the end (100.7 +/- 19.3 ppm x hours) of the 21-day period. Gastrointestinal symptoms were negligible during both the breath hydrogen tests as were symptoms during the 21-day period. APPLICATIONS/CONCLUSIONS: The diet was well tolerated by the subjects. Furthermore, the decrease in breath hydrogen suggests colonic adaptation to the high-lactose diet. The results indicate that lactose maldigestion should not be a restricting factor in developing adequate calcium diets for this population. The existence of lactose maldigestion does not result in lactose intolerance in this population when it is fed a dairy-rich diet. PMID- 10812377 TI - Use of low-fat foods by people with diabetes decreases fat, saturated fat, and cholesterol intakes. AB - OBJECTIVE: To investigate the effect of providing free access to several fat modified foods on dietary energy and macronutrient intake in people with and without diabetes mellitus. DESIGN: Five low-fat or no-fat products or their regular-fat counterparts were provided to volunteers to take home and use for 3 days (low-fat condition or regular-fat condition) in a repeated-measures crossover design. People with diabetes were case matched to people without diabetes. Food intakes were determined through a weighed food diary and by weighing the food provided before consumption and the uneaten portions after consumption. SUBJECTS: Thirty men and women, aged 20 to 60 years, with (n = 15) and without (n = 15) diabetes participated. STATISTICAL ANALYSES: Repeated measures analysis of variance was used to determine the effects of diabetes and use of fat-modified foods on nutrient and energy intake. RESULTS: People with diabetes responded the same way to fat-modified foods as people without diabetes. There was a significant reduction in the grams of fat consumed during the low-fat condition compared with the regular-fat condition (average decrease = 8 g, P < .05). Energy intake from experimental foods was significantly lower during the low-fat condition (271 +/- 181 kcal) compared with the regular-fat condition (353 +/- 256 kcal), but total energy intake was not different. Percentage of energy from fat was significantly decreased in the low-fat condition (27 +/- 7) compared with the regular-fat condition (34 +/- 9; P < .05). There was a corresponding increase in the percentage of energy from carbohydrates in the low-fat condition compared with the regular-fat condition, but no significant increase in grams of carbohydrate consumed. Cholesterol and saturated fat intakes were significantly less in the low-fat condition than in the regular-fat condition. CONCLUSION: Consumption of fat-modified foods by individuals with diabetes may help decrease intake of fat, cholesterol, and saturated fat. PMID- 10812378 TI - Diet high in monounsaturated fat does not have a different effect on arterial elasticity than a low-fat, high-carbohydrate diet. AB - OBJECTIVE: To compare the effects of a modified-fat diet high in monounsaturated fat, and a low-fat/high-carbohydrate diet on arterial elasticity. DESIGN: Randomized crossover design; each diet period was 1 month and a 2-week wash out period occurred in between. SUBJECTS/SETTING: Thirty healthy, free-living, nonsmoking men and women were recruited from the Melbourne, Australia, metropolitan region of Australia. Men were aged 35 to 55 years and postmenopausal women were aged 50 to 60 years and were not taking hormone replacement therapy. Twenty-eight subjects completed the study. INTERVENTION: Two diets of equal energy value: a modified-fat diet and a low-fat/high-carbohydrate diet; the modified-fat diet had 3 times more energy from monounsaturated fat. MAIN OUTCOME MEASURES: Arterial elasticity and serum lipoprotein concentrations. STATISTICAL ANALYSIS: The general linear model was used to investigate overall effect and any carryover or order effects. Paired t test and the general linear model were used to compare the results from the 2 diet periods. RESULTS: High-density lipoprotein cholesterol concentration was significantly higher on the modified-fat diet than on the low-fat/low-carbohydrate diet. Arterial elasticity and concentrations of total cholesterol, low-density lipoprotein cholesterol, and triglycerides were not significantly different on the 2 diets. APPLICATIONS/CONCLUSIONS: There is no evidence to favor a diet high in monounsaturated fat over a low-fat/high carbohydrate diet because of an effect on arterial elasticity. Other changes in diet may be needed to cause a beneficial effect on arterial elasticity. PMID- 10812379 TI - The knowledge, attitudes, and practices of dietitians licensed in Oregon regarding functional foods, nutrient supplements, and herbs as complementary medicine. AB - OBJECTIVE: To examine the perceived knowledge and attitudes of dietitians licensed in Oregon (LDs) regarding the effectiveness and safety of functional foods, nutrient supplements, and herbs as complementary medicine as well as their personal use, recommendations for the use of others, and training needs. DESIGN: A mailed survey was used to gather data. The questionnaire was developed and face validated after a focus group discussion. SUBJECTS/SETTING: A geographically stratified, random sample of 202 Oregon LDs was surveyed; usable data were collected from 162 LDs (80%). STATISTICAL ANALYSES: Descriptive statistics and chi 2 tests were used to analyze data. The data were weighted to take account of the sampling method and yield population estimates. RESULTS: LDs considered themselves to be knowledgeable of functional foods and nutrient supplements. More than 80% were confident of the effectiveness of functional foods and nutrient supplements for prevention of illness and treatment of chronic illness, and at least 89% were confident of their safety for these uses. Fewer than 75% considered herbs to be safe and only about 50% were confident of the effectiveness of herbs. Only 10% or fewer LDs considered themselves to be knowledgeable about herbs for prevention and treatment of illness. Interest in training about each of these less-traditional nutrition therapies was high. APPLICATIONS/CONCLUSIONS: Oregon LDs consider themselves to be actively practicing in the area of complementary medicine and have a strong desire for further training. The dietetics profession can capitalize on this interest by taking the lead in training dietitians about the role of functional foods, nutrient supplements, and herbs in health. PMID- 10812380 TI - Nutrient contribution of infant cereals used as fluid thickening agents in diets fed to the elderly. AB - OBJECTIVE: To evaluate whether a nutrient-fortified fluid-thickening agent (Pablum, H. J. Heinz Co of Canada, North York, Ontario, Canada) replaces nutrients lost to food displacement associated with its use. DESIGN: Seven-day, investigator-weighted, food intake records were evaluated to determine the prevalence of nutrient inadequacy. Nutrient intakes, including and excluding those associated with Pablum, were assessed to determine the ability of Pablum to protect from nutrient inadequacy. SUBJECTS/SETTING SUBJECTS: (19 women and 2 men aged 69 to 109 years) were residents of a home for the aged or a chronic-care hospital who required pureed food and thickened fluids. STATISTICAL ANALYSES PERFORMED: Probability analysis was used to estimate the prevalence of nutrient inadequacy for micronutrients and protein. RESULTS: Approximately 15% of consumed energy (mean +/- standard deviation: 1.534 +/- 310 kcal/day; 1.38 +/- 0.37 and 1.46 +/- 0.26 multiplied by basal metabolic rate for men and women) came from the thickener. If a nonfortified thickener was used, risk of inadequacy (percentage of sample) would be apparent for protein (16%), calcium (95%), thiamin (57%), riboflavin (28%), niacin (55%), and folate (47% for women and 97% for men); no risk was estimated for iron and vitamins A and C. The nutrients contained in Pablum reduced or eliminated the risk of inadequacy for some nutrients, including protein (8%), calcium (9%), thiamin (0%), riboflavin (0%), and niacin (0%). In contrast, inadequate consumption of water and folate occurred even when the contribution of Pablum was considered. APPLICATIONS: The use of a nutrient fortified thickening agent has merit; however, the current formulation of Pablum does not allow for complete protection against nutrient inadequacy. PMID- 10812381 TI - Design, development, and formative evaluation of "put nutrition into practice," a multimedia nutrition education program for adults. AB - The purpose of this study was to design, develop, and formatively evaluate a computer-based multi-media nutrition education program for adults based on the Dick and Carey model of instructional design. The 4 phases of the study included analysis, design, development, and evaluation. Seventy-two volunteers from the US Air Force, aged 18 to 50 years, participated in focus groups, an E-mail survey, or a dietitian survey to establish the program's instructional goal of applying the principles of the Food Guide Pyramid to daily food choices. Objectives, assessment instruments, content, examples, and practice questions with feedback were written in the design phase. Four modules of instruction--Familiarization with Food Groups, Serving Sizes, Modifying a Menu, and Vitamins and Minerals- were programmed using Hyper-studio. Eighteen subjects aged 22 to 40 years, with at least a high school education and an average knowledge of nutrition volunteered to participate in 1 of 2 formative evaluation phases. All subjects completed a pretest, 2 posttests, 3 embedded tests, and an attitude questionnaire to ascertain program weaknesses. One module was deleted after phase 1 because the material lacked relevance to subjects. In phase 2, only 4 of 15 subjects could identify serving sizes in module 2 and only 6 of 15 subjects could do the same on posttest 1. Back buttons and review screens were added to modules 2 and 3 to facilitate identification of serving sizes. We conclude that dietetics professionals should use systematic models of instructional design, such as the Dick and Carey model, to design effective nutrition education programs for the public. PMID- 10812382 TI - Preventing obesity in American Indian children: when to begin. PMID- 10812383 TI - Behaviors of low-income mothers related to fruit and vegetable consumption. PMID- 10812384 TI - Blood glucose laboratory for first-year medical students: an interdisciplinary model for nutrition-focused diabetes management. PMID- 10812385 TI - Free galactose content of fresh fruits and strained fruit and vegetable baby foods: more foods to consider for the galactose-restricted diet. PMID- 10812386 TI - Do consumers of savory snacks have poor-quality diets? PMID- 10812387 TI - Position of the American Dietetic Association: nutrition, aging, and the continuum of care. AB - Scientific evidence increasingly supports that good nutrition is essential to the health, self-sufficiency, and quality of life of older adults. With the population of the United States living longer than ever before, the older adult population will be more diverse and heterogeneous in the 21st century. The oldest old and minority populations will grow more quickly than the young-old and non Hispanic white populations, respectively. For the current 34 million adults 65 years of age and older living in the United States, there are about 12 million caregivers who provide formal or informal care. A broad array of culturally appropriate food and nutrition services, physical activities, and health and supportive care customized to accommodate the variations within this expanding population of older adults is needed. With changes and lack of coordination in health care and social-support systems, dietetics professionals need to be proactive and collaborate with aging-services and other health care professionals to improve policies, interventions, and programs that service older adults throughout the continuum of care to ensure nutritional well-being and quality of life. The American Dietetic Association supports both the provision of comprehensive food and nutrition services and the continuation and expansion of research to identify the most effective food and nutrition interventions for older adults over the continuum of care. PMID- 10812388 TI - Preoperative radiation with concurrent 5-fluorouracil for locally advanced T4 primary rectal cancer. AB - PURPOSE: In cT4-rectal carcinoma disease-free margins often cannot be obtained by primary surgery, and even if total en bloc resection is accomplished, local failure remains high with surgery alone. Herein we report on the curative resectability rate, acute toxicities, surgical complications, local control and 5 year survival rates achieved with a more aggressive multimodality regimen, including preoperative radiochemotherapy. PATIENTS AND METHODS: Between 1/1990 and 12/1998, a total of 31 patients with cT4-rectal cancer were treated at our institution. All patients presented with tumor contiguous or adherent to adjacent pelvic organs. Eight patients had synchronous distant metastases. A total radiation dose of 50.4 Gy with a small-volume boost of 5.4 to 9 Gy was delivered (single dose: 1.8 Gy). 5-FU was scheduled as a continuous infusion of 1000 mg/m2 per 24 hours on days 1 to 5 and 29 to 33. Six weeks after completion of radiochemotherapy, patients were reassessed for resectability. RESULTS: After preoperative radiochemotherapy, 29/31 patients (94%) underwent surgery with curative intent. Resection of the pelvic tumor with negative margins was achieved in 26/31 patients (84%), 3 patients had microscopic residual pelvic disease. In 3/8 patients with distant spread at presentation a complete resection of metastases was finally accomplished. Toxicity of radiochemotherapy occurred mainly as diarrhea (NCI-CTC Grade 3: 23%), dermatitis (Grade 3: 16%) and leucopenia (Grade 3: 10%). Surgical complications appeared as anastomotic leakage in 3, wound infection in 2, fistula, abscess and hemorrhage in 1 patient, respectively. With a median follow-up of 33 months, local failure after curative resection was observed in 4 patients (19%), 3 patients (14%) developed distant metastases. The 5-year overall survival rate for the entire group of 31 patients was 51%, following curative surgery 68%. CONCLUSION: A combination of high-dose preoperative radiochemotherapy followed by extended surgery can achieve clear resection margins in more than 80% of patients with locally advanced cT4 rectal tumor. An encouraging trend evolves for this multimodality treatment to improve long-term local control and survival. PMID- 10812389 TI - [Prostatic carcinoma: problems in the interpretation of rectal dose-volume histograms]. AB - BACKGROUND: Dose-volume histograms (DVHs) are used for the prediction and calculation of late radiation side effects. In literature the predictive value of rectal DVHs is controversially discussed. Differences in contouring might contribute to the contradicting results. In particular the cranial and caudal border of the contoured organ are not uniformly defined. PATIENTS AND METHODS: The DVHs of 12 patients who were treated with conformal radiotherapy for prostate cancer were investigated. Six of the patients suffered from mild rectal bleeding as a late side effect of radiotherapy. Six patients without rectal bleeding (minimal follow-up 30 months) matched for age, concomitant disease and treatment concept served as controls. Four different DVHs with 4 different definitions of the cranial and caudal rectal border were generated for each patient. For each of the 48 DVHs the percent volume fractions (V50, V80, V95) and absolute volume fractions (aV50, aV80, aV95) were calculated that received more than 50%, 80% and 95% of the reference dose. RESULTS: For every patient there were considerable variations in the volume fractions depending on the definition of the rectum borders (Table 1). The mean and median values of the percent and absolute volume fractions of the bleeding patients were higher than those of the non-bleeders no matter how the rectum borders were defined. None of the volume fractions could totally separate bleeding from non-bleeding patients. CONCLUSION: There is a high variability of absolute and percent volume fractions of rectal DVHs depending on how the rectal borders were defined. For the comparison and for the interpretation of rectal DVHs a uniform definition would be helpful. PMID- 10812391 TI - Evaluation of the extension of cerebral gliomas by scintigraphy. AB - BACKGROUND: Single photon emission computed tomography (SPECT) with 201Tl and 123I-alpha-methyl tyrosine (123I-IMT) are routine methods for the evaluation of brain tumors. 123I-IMT transport across the blood brain barrier is mediated by an amino acid carrier, 201Tl accumulation is analogous to cerebral potassium uptake. PATIENTS AND METHODS: To determine the differences in glioma extension as shown by the 2 methods, 17 patients with malignant gliomas were included in this comparative imaging study: astrocytoma III: n = 6, ependymoma III: n = 1, oligodendroglioma III: n = 1, glioblastoma IV: n = 9. The tomographic image sets were matched anatomically and the slices showing maximal tumor extension were identified in both image sets respectively. Tumor spread was compared visually and the tumor extension was quantified. RESULTS: In gliomas WHO III tumor extension was delineated significantly larger by 123I-IMT-SPECT than by 201Tl SPECT (mean +/- SD: 816 +/- 281 pixels vs 600 +/- 220 pixels, n = 8, p < 0.05). The size of glioblastomas was shown in a comparable manner by the 2 methods (977 +/- 571 vs 1.051 +/- 588, n = 9, ns, p = 0.57), but there were considerable regional differences between the area of 201Tl uptake and amino acid retention. In the whole group a weak but significant negative correlation between intensity of 201Tl uptake on the one hand and a ratio of the area as depicted by 123I-IMT vs area as depicted by 201Tl on the other hand, was found (n = 17, r = 0.49, p < 0.05). Thus the differences in the delineation of areas became smaller with increasing 201Tl uptake. CONCLUSIONS: These preliminary data indicate that the extension of gliomas is depicted differently by the 2 methods. 123I-IMT-SPECT shows a larger tumor extension especially in gliomas WHO III. Since 201Tl uptake has previously been shown to correlate with disruption of the blood brain barrier, 123I-IMT-SPECT may delineate tumor parts without endothelial leakage. This additional information may be helpful in planning surgical or radiation therapy. The advantages of 123I-IMT in this respect decrease with increasing 201Tl uptake and with increasing malignancy. PMID- 10812390 TI - [Prevention of radiation-induced diarrhea by smectite. Results of a double-blind randomized, placebo-controlled multicenter study]. AB - PURPOSE: Diarrhea and abdominal pain are well-known side effects abdominal or pelvic of radiation therapy that may lead to interruption of treatment in serious cases. In recent trials the silicate smectite has proven a promising drug in the prophylaxis of these adverse events. The presented trial aimed at the verification of earlier studies and the evaluation of a dose-effect relationship. PATIENTS AND METHODS: Between April 1994 and May 1995, a total of 176 patients obtaining radiotherapy of the pelvis or the abdomen were evaluated in a double blind, randomized placebo-controlled investigation regarding the prophylactic effect of smectite (= Colina) against radiotherapy-induced diarrhea. During the whole period of radiotherapy 85 patients obtained 2 x 6 g smectite daily and 91 patients received 2 x 6 g placebo. The primary end point of the analysis was the time to the first appearance of diarrhea (> or = 3 pappy stools). RESULTS: All 176 patients were evaluated according to an intent-to-treat analysis. There was no significant difference between the prophylactic effects of smectite and placebo. For an explorative post-hoc analysis the total study group was split up into 2 subgroups, one with an irradiated small bowel volume < or = 837.5 ml, the other with a small bowel volume > 837.5 ml (median); the analysis indicated that the first subgroup showed a benefit for the smectite-treated patients in contrast to the placebo treatment (32 vs. 18 calendar days to the first appearance of diarrhea). This benefit was statistically not significant. CONCLUSION: Prophylactic application of smectite during irradiation of the pelvis and the abdomen can delay the development of radiotherapy-induced diarrhea, a statistical significance could not be verified neither in the total study group nor in the post-hoc subgroup analysis. PMID- 10812392 TI - Influence of apoptosis on the enhancement of radiotoxicity by ouabain. AB - BACKGROUND: The Na+, K(+)-ATPase inhibitor ouabain enhances the toxicity of irradiation and we have previously demonstrated that the drug suppresses repair capacity. The influence of ouabain on apoptosis is not known and is examined in this study. MATERIALS AND METHODS: Seven human cell lines of defined TP53 status were irradiated with 60Co-gamma irradiation in the presence and absence of 10(-7) M ouabain. Cell survival was determined by the clonogenic assay, apoptosis by acridine orange staining and cell cycle delays by flow cytometry. RESULTS: The ouabain-induced enhancement of radiotoxicity, expressed as the ratio of SF2's, is independent of TP53 status and ranges from 1.1 to 2.8 depending upon cell line. Ouabain prolongs the irradiation-induced G2 delay in TP53 mutant tumor cell lines by a factor greater than 2, but not in the normal lung fibroblast L132, where the cell recovery is not altered in the presence of ouabain. Twenty hours post irradiation, ouabain enhances apoptosis induced by irradiation by factors of 1.3 to 1.7 depending on the cell line. CONCLUSION: Ouabain preferentially enhances the radiotoxicity in tumor cells irrespective of TP53 status. In the pattern of DNA damage responses which are influenced by ouabain we show that the G2 cell cycle delay is prolonged and that early apoptosis events are upregulated in TP53 wild type and TP53 mutant cells. It is concluded that apoptosis plays a significant role in the enhancement of radiotoxicity by ouabain. PMID- 10812393 TI - Diagnostic radiation oncology: malignant cystosarcoma phylloides. AB - BACKGROUND: Cystosarcoma phylloides is a rare, mostly benign tumor of the breast. In most cases curative treatment is possible with complete surgical excision of the lesions. However, there is a high local recurrence rate up to 46%. There are only few reported successful treatments with radiotherapy in cystosarcoma phylloides. Radiotherapy is indicated in patients with locoregional recurrent disease and those with symptomatic metastases. PATIENT: We present a case of a 54 year-old woman with a painless mass in the right breast. Clinical examination showed a large, firm tumor and axillary lymph node metastases. The patient underwent mastectomy and axillary lymph node dissection. Histopathology showed a cystosarcoma phylloides measuring 30 x 25 x 19 cm as well as lymph node metastases measuring 9.5 cm. Despite a R0 resection the patient developed 4 thoracic wall recurrences within 2 years. At 18 months there were additional multiple pulmonary and pleural lesions with a maximum diameter of 6.5 cm. Subsequently the patient received polychemotherapy. There was a significant response of the thoracic wall disease as well as of the pulmonary and pleural lesions. Six months later there was further progress of the thoracic wall disease and the pulmonary lesions. Following this the patient underwent radiation therapy of her right thoracic wall with 50 Gy which lead clinically to a quick response. For the past 5 months there has been no evidence of thoracic wall disease recurrence. CONCLUSION: This is an unusual case of cystosarcoma phylloides and response to different therapeutic modalities. Cystosarcoma phylloides showed to be a radiosensitive tumor. Thus postopertive radiation therapy is indicated to prevent local recurrences. PMID- 10812394 TI - [Consolidation radiotherapy after high-dose chemotherapy and autologous bone marrow transplantation in patients with advanced breast cancer]. PMID- 10812395 TI - [Long-term cardiac sequelae of adjuvant radiotherapy after mastectomy? Analysis of Danish DBCG-82b/82c trials]. PMID- 10812397 TI - [In Process Citation] PMID- 10812396 TI - [Quality of life after simultaneous radiochemotherapy in squamous cell carcinoma of the head and neck]. PMID- 10812398 TI - Multiple crusted cutaneous plaques in a cat. PMID- 10812399 TI - Polycystic kidney disease in bull terriers: an autosomal dominant inherited disorder. AB - The prevalence, mode of inheritance and urinalysis findings in Bull Terriers with polycystic kidney disease were assessed by screening 150 clinically normal dogs. The disorder was diagnosed in 39 dogs on the basis of renal ultrasound results and family history of the disease. In equivocal cases confirmation required gross and histopathological renal examination. Necropsy was performed on nine affected dogs and the kidneys from another five affected animals were also examined. Renal cysts were usually bilateral, occurred in cortex and medulla and varied from less than 1 mm to over 2.5 cm in diameter. Cysts were lined by epithelial cells of nephron origin. Abnormal urine sediment and proteinuria were common in affected dogs. The disease appears to be inherited in a highly penetrant autosomal dominant manner. PMID- 10812400 TI - Mycobacterial infection in an ostrich. PMID- 10812401 TI - Treatment of progressive ethmoidal haematoma using intralesional injections of formalin in three horses. AB - Three Thoroughbred horses with unilateral progressive ethmoid haematomas were treated using intralesional injections of 10% formalin (4% formaldehyde solution). Injections were performed in the standing sedated horse through the nasal passages under endoscopic guidance and, when the ethmoid haematoma involved the paranasal sinuses, through holes trephined into the affected sinus. Regression of the lesions occurred in all cases after repeated injections. This technique appears to be a safe and effective treatment for progressive ethmoid haematomas in the horse. PMID- 10812402 TI - Management of perianal fistulae in five dogs using azathioprine and metronidazole prior to surgery. AB - OBJECTIVE: To evaluate combination therapy with azathioprine and metronidazole in German Shepherd Dogs with perianal fistulae. DESIGN: Prospective study. PROCEDURE: Five dogs (31.5 to 36.0 kg) with perianal fistulae were treated with azathioprine (50 mg per dog orally every 24 h) and metronidazole (400 mg per dog orally every 24 h). Patients were re-evaluated at 2 week intervals by inspection, palpation, photographs of the perineal region and assessment of white blood cell counts where possible. Treatment was continued until improvement in lesions reached a plateau. Surgical excision of residual fistulae and anal sac remnants was then performed, with medical therapy continued for an additional 3 to 6 weeks. RESULT: Signs attributable to anal irritation were reduced or eliminated in all dogs within 2 weeks, although visible healing of lesions progressed more slowly. Ulcerated lesions reduced in surface area and depth, and some fistulae healed completely. Non-healing areas were usually associated with anal sac rupture or chronic fibrosis. Visible improvement typically reached a plateau 4 to 6 weeks after commencing treatment. Immunosuppressive therapy continued for 5 to 24 weeks before surgical intervention to remove anal sacs (four dogs) and/or residual fistulae (five dogs). All dogs remain disease free 7 to 10 months postoperatively. No important complications of treatment were encountered. CONCLUSION: Azathioprine with metronidazole effectively reduced perianal irritation, and the severity and extent of lesions prior to surgery. Treatment was economical even in large dogs and associated with few untoward sequelae. The combined use of immunosuppressive and antimicrobial therapy followed by surgery minimised potential morbidity associated with aggressive use of either medical of surgical treatment alone. PMID- 10812403 TI - A moment in veterinary history--1888 to 1998. PMID- 10812404 TI - The arts in the lives of Australian veterinarians: a preliminary survey. AB - OBJECTIVE: To assess the role of the arts in the lives of those veterinary students and veterinarians who could be found to be active in one or more of the arts. PROCEDURE: Veterinary students and veterinarians active in the arts were sought by advertisement and personal contact. Respondents each completed a questionnaire, which was analysed using SAS. RESULTS: Of the 55 respondents (43 veterinarians; 12 students), 73% were involved in music, 31% in theatre, 27% in writing and 15% art, but for more than 80% this occupied no more than 10 hours/week, about one quarter of the time for veterinary work. Although most would like to spend more time on the artistic activity, this was generally limited by the need to earn an income from veterinary work. Music and drama provided satisfaction, variety, and relaxation while catering to creative needs. A creative outlet and satisfaction was very important for writers as well, but they also valued the intellectual stimulation. A complex relationship existed between the artistic and the veterinary activities, but the nature of this relationship varied both between type of artistic activity, and the person concerned. CONCLUSIONS: Veterinary science and the arts can complement one another with each making important contributions to the person and the artistic field involved, even though limitations of time for the arts may cause frustration. PMID- 10812405 TI - Variation of pesticide concentration in sheep dips operated according to traditional and revised methods. AB - OBJECTIVE: To quantify stripping in traditional dipping operations and to revise dipping methods, based on prediction of stripping so that a more stable concentration of pesticide in the dipwash is achieved. DESIGN AND METHODS: Plunge and shower dips were operated sequentially according to traditional and revised dipping instructions. Dips were operated by continuous and intermittent replenishment. Samples of mixed dipwash were collected periodically and assayed for pesticide (diazinon) concentration. RESULTS: Diagrammatic representations of pesticide concentration versus number of sheep dipped indicated traditional dipping leads to wide variations in the concentration of pesticide in dipwash during dipping. Intermittent replenishment led to a 'saw-tooth' pattern in the pesticide concentration. Traditional continuous replenishment (using the starting concentration of pesticide) indicated both the rate and extent of stripping was higher in shower dipping. If sufficient sheep were dipped, equilibrium was reached between the rate of pesticide replenishment and removal. An alternative method of dip operation by continuous replenishment, using a low starting concentration of pesticide and a replenishment concentration high enough to offset the pesticide loss through stripping resulted in a more stable concentration of pesticide in the dip. CONCLUSION: Revision of dipping instructions can lead to exposure of sheep to stable concentrations of stripping pesticide during dipping. PMID- 10812406 TI - Comparative immunogenicity of two bivalent botulinum vaccines. AB - OBJECTIVE: To compare the ability of a new single-dose botulinum vaccine containing a non-mineral oil adjuvant with a single dose of a conventional botulinum vaccine product to produce antibody to Clostridium botulinum types C and D in cattle in Northern Australia. DESIGN AND PROCEDURE: One hundred and fifty Brahman steer weaners were randomly divided into two groups receiving either a single dose of CSL Bivalent Botulinum vaccine or Websters Singvac. Blood samples were collected at 0, 8 and 24 weeks and tested by antibody ELISA. The final samples were also tested by the toxin neutralisation test, to test titres of neutralising antibody. RESULTS: Six months after inoculation, cattle vaccinated with Websters Singvac had ELISA antibody response twice that of CSL conventional product. However, this difference was only evident for neutralising antibody to type C botulinum toxin. Both products produced similar titres of type D neutralising antibody after a single dose. CONCLUSION: Websters' Singvac produces a greater neutralising antibody response to type C botulism upon single inoculation than a conventional vaccine. The product produces an equivalent neutralising antibody response to type D. PMID- 10812408 TI - Incidental lesions in the brains of sheep and goats. PMID- 10812407 TI - Detection of Mycobacterium avium subsp paratuberculosis in formalin-fixed paraffin-embedded intestinal tissue by IS900 polymerase chain reaction. AB - OBJECTIVES: To evaluate and compare methods for DNA extraction from formalin fixed, paraffin-embedded tissues and methods for detection of Mycobacterium avium subsp paratuberculosis by IS900 PCR for confirmation of Johne's disease in ruminants. DESIGN: A laboratory study. PROCEDURE: Three methods of DNA extraction of differing complexity and two PCR protocols using different pairs of IS900 primers were compared. Sensitivity and specificity were assessed using samples from ruminants with and without histological evidence of Johne's disease. RESULTS: The simplest method of DNA extraction, which involved two cycles of boiling and freezing followed by centrifugation, gave more consistent results than two methods that required solvent extraction of paraffin, proteinase digestion and DNA purification. The sensitivity of detection of M avium subsp paratuberculosis in paraffin blocks stored for 1 to 6 years from 34 cases of Johne's disease in sheep, cattle and goats was 88% for a 229 bp IS900 PCR assay and 71% for a 413 bp assay, using the detection of acid-fast bacilli by Ziehl Neelsen staining of histological sections from the same blocks as the gold standard test. PCR results correlated with the abundance of acid-fast organisms in the tissues. No false positive reactions were detected. CONCLUSION: PCR for identification of M avium subsp paratuberculosis in formalin-fixed, paraffin embedded intestinal tissues from ruminants is a rapid and useful method. A simple method of sample preparation is effective. Amplification of short fragments of IS900 is more effective than amplification of longer fragments. PMID- 10812409 TI - Psammomatous cerebral intraventricular meningioma in a bull. PMID- 10812410 TI - Pyrrolizidine alkaloid poisoning of horses grazing a native heliotrope (Heliotropium ovalifolium). PMID- 10812411 TI - Preanesthetic evaluation of the cancer patient. PMID- 10812412 TI - Anesthetic considerations in radiologic procedures performed outside the operating room. PMID- 10812413 TI - Anesthetic management of the patient undergoing head and neck cancer surgery. AB - The head and neck cancer patient should be in the best possible medical condition before facing surgery, bearing in mind the status of the tumor and the urgency of the procedure. Careful assessment of the patient's upper airway will enable the anesthesiologist to select an appropriate course of action to secure the airway before the operation begins. In many cases, the patient can be safely intubated after the induction of general anesthesia. In other situations, the patient may require an examination of the airway while awake with the aid of sedation and topical analgesia to determine the safest intubation technique. If the patient has evidence of a difficult airway, a flexible fiberoptic-guided intubation may be indicated to secure the airway in the awake patient patient before general anesthesia is induced. Some patients with severe airway obstruction or large, bulky supraglottic tumors usually undergo an initial tracheostomy with local anesthesia to secure the airway. Following surgery, extubation of the patient's trachea requires careful attention and may have to be performed over a jet ventilating stylet. PMID- 10812414 TI - Anesthetic implications for surgical patients with endocrine tumors. AB - The preoperative evaluation and safe anesthetic treatment of patients with endocrine gland tumors mandate an understanding of the physiologic dysfunctions attributable to these tumors. Some patients will exhibit various signs and symptoms characteristic of the MEN syndromes. In the patient with acromegaly, a fiberoptic-guided intubation of the trachea to secure the airway before induction of general anesthesia must be anticipated. Anesthetic treatment of the patient with hyperadrenocorticism requires knowledge of the physiologic effect of excess cortisol. In the patient with severe hyperparathyroidism, we attempt to correct the markedly elevated plasma calcium levels and maintain adequate hydration and urine output perioperatively. Following thyroidectomy for MCT, 2 potential problems of concern are upper airway obstruction and aspiration resulting from injury (unilateral or bilateral) to the recurrent laryngeal nerve and the superior laryngeal nerve, respectively. The major focus during excision of an insulinoma is prevention of wide swings in blood glucose concentrations. In the gastrinoma patient, the anesthesiologist not only must correct any intravascular fluid volume deficit or electrolyte imbalance but must also consider the patient to have a full stomach at the time of anesthetic induction. Correction of hypokalemia and control of hypertension may be required in the preoperative preparation of the patient with an adrenal cortex tumor. Preoperative alpha adrenergic blockade must be initiated in the patient with a pheochromocytoma to prevent dangerous elevations in blood pressure during anesthesia and surgery for the tumor's removal. Vasodilators with rapid onset and short duration are used to treat intraoperative hypertension. After ligation of the tumor's blood supply, falls in blood pressure may require treatment with fluids and vasopressors. Carcinoid syndrome patients should be treated with somatostatin to prevent stimuli such as anxiety, abdominal scrubbing, or tumor manipulation from precipitating severe hypotension, hypertension, bronchospasm, or tachycardia. In both pheochromocytoma and carcinoid patients, a smooth anesthetic induction and tracheal intubation plus avoidance of drugs that release histamine or activate the sympathetic nervous system may also prevent intraoperative crises. PMID- 10812415 TI - Anesthetic management of the cancer patient undergoing noncardiac thoracic surgery. AB - The scope of this article is necessarily limited, but we have addressed preoperative assessment, monitoring, and intraoperative support that might be required for surgical patients with lung, mediastinal, and esophageal cancer. The development of the subspecialty of thoracic anesthesia has been responsible for reducing mortality and improving care of all patients undergoing noncardiac thoracic surgery and has been one of the forces behind the surgeon's ability to handle bigger challenges with better results. PMID- 10812416 TI - ARDS in the postoperative cancer patient. PMID- 10812417 TI - Postoperative pain management. AB - Postoperative pain can be effectively managed, even in the most complex oncologic procedures. Although the primary agents for treatment of severe pain continue to be opioids, routes of administration and dosing regimen have undergone a dramatic metamorphosis in the past 10 years. The intramuscular injection given every 4 hours has been replaced by patient-controlled analgesia and epidural techniques. Management of ancillary issues that contribute to an increased perception of pain (i.e., stress, depression, anxiety, and inflammation) must be included in an effective multimodal plan. Closer attention to the treatment of pain can obviate the consequences of poorly managed pain, which we are only beginning to understand. In this day of active consumerism in medicine, patients have come to expect improved pain management. Early outcome studies are beginning to confirm the belief that improved pain management translates into between outcomes and earlier dismissals. In the first century BC, Publilius Syrus, a Latin mime, wrote, "There are some remedies worse than the disease." For centuries, pain was inextricably linked to treatment. We may now be approaching a time in the development of medical care when this is no longer true. PMID- 10812418 TI - Neural blockade for abdominopelvic pain of oncologic origin. AB - Neural blockade, like other accepted treatments for persistent pain, is not a panacea. Careful assessment is required to elicit the need for complementary interventions, including pharmacological management and psychobehavioral and rehabilitative approaches, combined with attention to the palliation of other symptoms. Celiac and superior hypogastric plexus blocks are well-accepted, effective, and minimally hazardous means for providing palliation of visceral abdominopelvic pain. Although they require radiological imaging, they are relatively undemanding of the experienced anesthesiologist pain specialist and do not deplete patients' limited resources and energy. Because of their uniquely favorable risk:benefit ratio, these procedures should be considered early in the course of treating patients with abdominopelvic pain that is expected to persist. PMID- 10812419 TI - Intraspinal opioid therapy for intractable cancer pain. PMID- 10812420 TI - Anesthetic management of the patient who refuses blood transfusions. PMID- 10812421 TI - Ethical issues in surgical oncology patients. PMID- 10812422 TI - Risks and outcomes in oncology patients undergoing anesthesia and surgery. PMID- 10812423 TI - Characterization of a sapphire-epoxy coating for capillary electrophoresis. AB - A new procedure for coating capillaries for capillary electrophoresis applying a sapphire (alumina) containing epoxy resin was developed. Coated capillaries showed considerably reduced electroosmotic flow, and decreased the adsorption of proteins to the internal wall of the capillary. Coating is transparent down to 195 nm and can be used with advantage to analyze different kinds of substances, such as small cations and/or anions, and proteins. PMID- 10812424 TI - Meat species identification by linear discriminant analysis of capillary electrophoresis protein profiles. AB - The objective of this study was to utilize linear discriminant analysis (LDA) in the interpretation of capillary electrophoresis-sodium dodecyl sulfate polymer filled capillary gel electrophoresis (CE-SDS) meat protein profiles for the identification of meat species. The specific objectives were 1) to collect quantitative data on water-soluble and saline-soluble proteins of different meat species obtained by CE-SDS and 2) to apply LDA on collected CE-SDS protein data for the development of a pattern recognition statistical model useful in the differentiation of meat species. Samples were raw beef top and eye round, boneless fresh pork ham and loin, turkey leg and breast meat, and mechanically deboned turkey meat collected on six different occasions, making a total of 42 samples. Additionally, 14 samples were used as test samples to determine the classification ability of the procedure. Quantitative protein data obtained by CE SDS was used to generate separate LDA models for either water- or saline-soluble protein extracts. Although a saline solution was a more efficient meat protein extracting agent, as shown by a higher total protein concentration and a larger number of peaks, water-soluble CE-SDS protein profiles gave more distinctive discrimination among meat species. The correct classification given by LDA on water-soluble protein data was 100% for all meat species, except pork (94%). Conversely, the correct classification on saline-soluble protein data was 88% for beef and mechanically deboned turkey meat, and 94% and 100% for turkey and pork meat, respectively. LDA proved to be a useful pattern recognition procedure in the interpretation of CE-SDS protein profiles for the identification of meat species. PMID- 10812425 TI - Comparative analysis of storage proteins of Lotus spp. seeds by CGE and SDS-PAGE. AB - L. tenuis and L. corniculatus seeds are morphologically very similar but their purchase prices are quite different. Chromosome number counting is the only test used thus far in laboratories for the identification of these seeds. Recently, the flavonol's pattern has been used as a criterion for differentiation. In the present work, we studied the storage protein patterns of different Lotus seed samples by capillary gel electrophoresis (CGE), as an alternative method, comparing it with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). The seeds were treated according to International Seed Testing Association (ISTA) recommendations. CGE separations were performed using an uncoated capillary of 18 cm effective length and 50 microns i.d. and the Bio-Rad Protein Kit (Hercules, CA, U.S.A.). On-line detection was carried out at 220 nm. PMID- 10812426 TI - Study of major flavonoids in crude Scutellariae Radix by micellar electrokinetic capillary chromatography. AB - A method of micellar electrokinetic capillary electrophoresis for determining the six flavonoids in Scutellariae Radix (i.e., baicalin, baicalein, wogonin 7-O glucuronide, wogonin, oroxylin A 7-O-glucuronide, and oroxylin A) has been developed. The buffer solution (pH 7.24) composed of 20 mM sodium dodecyl sulfate (SDS), 20 mM sodium dihydrogen phosphate, and 25 mM sodium borate was found to be the most suitable electrolyte for the separation. The contents of the six flavonoids in crude Scutellariae Radix could easily be determined within about 15 min. On-column UV (254 nm) monitoring allowed the quantitative determination of baicalin. The effects of pH, surfactant concentration, and applied voltage on the migration behavior of the solutes were studied. PMID- 10812427 TI - Advances in detection of inorganic ions in capillary electrophoresis. AB - Recent advances in capillary electrophoresis methodology to improve the detection sensitivity for inorganic ions are briefly reviewed. Specifically discussed are current developments of nonabsorbance-based CE detectors and on-capillary enrichment techniques as well as some possible future perspectives. PMID- 10812428 TI - Characterization of humic substances by capillary electrophoresis. AB - Capillary electrophoresis was used for the separation of humic acid (HA) from peat, soil, and vermicompost. The electropherograms show the presence of at least three peaks eluted between 6 and 11 min for all HA. The best analysis resolution was obtained with the use of borate buffers at pH 8.9. The HA analyzed have structural and charge similarity, which increases the difficulty of separation. Therefore, the shape of the peaks is broad and the CE profiles of all HA are similar. It is reasonable to assume that the broad band in the three regions is due to the acidic groups that have a similar structure. By comparing the results obtained for HA extracted from soil, peat, vermicompost, and the commercial sample, HA from peat had the major carbon content. PMID- 10812429 TI - Capillary electrophoresis in China. AB - The history and present status of capillary electrophoresis in China are critically reviewed. Features of major research and future prospects are described. PMID- 10812431 TI - [Meeting of the French Society of Clinical Cytology]. PMID- 10812430 TI - A preliminary evaluation of serum proteins by capillary electrophoresis. AB - We evaluated the 5-band Serum Proteins by Capillary Electrophoresis kit (Bio-Rad Laboratories, Hercules, CA, U.S.A.) on the BioFocus 2000 CE (Bio-Rad) against conventional agarose gel electrophoresis (AGE) (Helena Laboratories, Beaumont, TX, U.S.A.). Serum from 60 patients was initially screened by AGE and divided into three groups: 1) normal electrophoretic pattern (n = 36, mean total protein 67.7 g/L), 2) monoclonal/oligoclonal gammopathy (n = 14, mean total protein 78.8 g/L), and 3) polyclonal gammopathy (n = 10, mean total protein 77.4 g/L). These samples were concurrently analyzed on the BioFocus 2000. Intraassay and interassay CVs for the five fractions for a normal sample were 0.17-1.44% and 0.42-9.11%, respectively, and 0.21-3.37% and 0.29-3.61%, respectively, for a sample with monoclonal gammopathy. Correlation coefficients for albumin and the albumin/globulin (A/G) ratio were 0.8891 (albumin range 17-93 g/L) and 0.8276 (A/G ratio range 0.39-7.81), respectively. The A/G ratio alone could not discriminate between the three groups. Capillary zone electrophoresis (CZE) correctly identified 33 of 36 samples in the normal group, and 22 of 24 samples in the other two groups, giving a clinical sensitivity and specificity of 91.7%. Our preliminary evaluation shows that protein separation by CZE is a simple, rapid, and automated alternative to conventional AGE. PMID- 10812432 TI - [Role of cytology and frozen sections in the intraoperative examination of the thyroid: comparison of two experiences]. AB - Two experiences of peroperative diagnosis in thyroid surgery are reported. In Bordeaux (France), frozen sections are supplemented by touch smears. Imprints alone give results similar to frozen but are not suitable in differentiating follicular adenoma from carcinoma; they appear more accurate for recognising the follicular presentation of papillary carcinoma. Touch smear is a rapid cost effective alternative to frozen section. In Brussels (Belgium), a more conventional attitude results in 185 frozen. A false negative ends in a follicular carcinoma. Eleven follicular proliferations will be signed out adenoma (8 cases) or carcinoma (3 cases) and three papillary carcinomas will only be detected after embedding. Six false negative derive from frozen section, no false positive is noted. An immediate change in the surgical procedure is justified twice. These observations argue for the free choice by the pathologist of the best technical procedure in peroperative diagnosis. PMID- 10812433 TI - [Intraoperative cytology of breast lesions]. AB - Evaluation of intraoperative cytology and frozen sections for breast lesions is essential to single-stage and cost effective management. The aim of this study is to evaluate the diagnostic accuracy and the potential role of intraoperative cytology. The results of the study undertaken at the Rene Huguenin Center and the data of the literature suggest that intraoperative cytology may be helpful in some cases, especially as an adjunct to frozen sections. If frozen sections could be avoided any time clear features of benignity or malignancy are offered by both clinical data and macroscopic and cytologic examination, this approach does have limitations. Intraoperative cytology should not be used as an alternative to frozen sections except, in a few cases, when technical conditions required for them are not available or suitable. It can also constitute a good way for continuous education of our cytotechnologists. PMID- 10812434 TI - [High grade (B) oligodendroglioma. Concerns and pitfalls in the intraoperative examination of smears]. AB - According to the Ste Anne hospital classification and grading system of oligodendrogliomas the presence of contrast enhancement on MRI and/or the presence of an histological endothelial hyperplasia are correlated with a pejorative prognosis (high grade or grade B). Surgical tumoral exerese and adjuvant chemo or radiotherapy are indicated. When clinical and radiological aspects are typical the extemporaneous diagnosis of oligodendroglioma on smear is usually easy (1). In case of atypical forms: unusual localization or cytological appearance, especially with a small cell hyperchromatic undifferenciated tumor few diagnostic hypothesismay be considered. Four cerebellar tumors are presented with relevant and differential cytological diagnostic criteria. A comparative study between clinical and radiological aspects of medulloblastoma, primitive lymphoma, undifferenciated small cell lung carcinoma, and grade B oligodendroglioma is presented. PMID- 10812435 TI - [Rapid staining and guided fine needle aspiration for deep masses: a focus]. AB - Ultrasound-, computerized tomography- or endoscopic ultrasonography-guided fine neddle aspiration (FNA) is a safe, rapid, and cost-effective method for securing a sample of abnormal tissue to diagnose and stage a variety of pathologic conditions in deep organs. The rate of false negative results is more dependent upon sampling failure and poor handling/preparation of aspirated material than on interpretation errors. This issue may be resolved in a cytopathologist is available at the time of the procedure to release a preliminary interpretation of air-dried, Diff-Quick-stained smears within a few minutes after performing the FNA. The immediate assessment can determine whether an adequate specimen is present, reduce the number of passes in each lesion, resulting in less discomfort and reduced likelihood of complications for the patient. A specific preliminary diagnosis may be rendered to guide further clinical investigation or treatment, and determine whether ancillary studies are needed to make a more accurate or specific diagnosis for the FNA specimen. Another advantage is the significant financial savings as compared to excisional tissue biopsy. But, rapid interpretation of the smears requires special training and is time-consuming. Accordingly, the indications have to be carefully determined, small-sized tumours are specifically recommended. A rapid evaluation increases the diagnostic yield, allowing near 100% in sensitivity, specificity and predictive value of positive cases. PMID- 10812436 TI - Cerebral amyloid angiopathy (CAA) with presentation as a brain inflammatory pseudo-tumour. AB - Cerebral amyloid angiopathy (CAA) is frequent but often asymptomatic. It can induce lobar haemorrhage, rapidly progressive dementia or recurrent transient neurological symptoms, other presentations being less frequent. We report 3 patients in their sixties presenting with a space occupying lesion which was the first manifestation of CAA. They were operated with a diagnosis of cerebral tumour. In all three cases, macroscopy was similar, the lesions were superficial in the cerebral cortex and the preoperative diagnoses were glioblastoma, meningioma and cavernoma. Histologically, the lesions consisted of a large inflammatory granuloma with numerous lipophages and siderophages surrounding capillaries with prominent endothelial cells. Vessels in the near cortex and meninges and within the granuloma harboured heavy amyloid deposits immunolabelled by anti-P component, anti-protein beta A4 with a A40 predominance and anti apolipoprotein E. Adjacent cerebral cortex showed reactive gliosis and rare senile plaques. Amyloidosis is rarely considered among diagnoses of space occupying lesions. In our three cases, CT scan and MRI changes were related to the presence of an inflammatory granuloma around foci of haemorrhage and amyloid laden vessels. PMID- 10812437 TI - Giant cell angiitis of the central nervous system with amyloid angiopathy. A case report and review of the literature. AB - We report a new case of giant cell angiitis of the central nervous system associated with cerebral amyloid angiopathy (GA/CAA). A 67-year-old woman was hospitalized with a history of headaches and lapses of consciousness. After improvement with corticosteroidtherpay, treatment was stopped. She relapsed and died 33 days after first admission. Pathological examination showed unusual extension of GA/CAA lesions, in the superficial and deep layer of the cerebral cortex, and in the cerebellum. Simultaneous occurrence of GA and CAA is rare. Histopathologic findings and immunological pathogenesis of the process are discussed: 1) arguments over pre-existence of CAA, responsible for GA; 2) primitive inflammatory process inducing amyloid deposits; 3) GA/CAA may represent an association of histological lesions related to 2 different types of disease: i) neurodegenerative disease with specific lesions (such as presence of diffuse senile plaques and neurofibrillary tangles) inducing inflammatory reaction ii) inflammatory disease, with few or no degenerative lesions, responding to immunotherapy. PMID- 10812438 TI - [Perceptual disorders in a Turkish woman: hallucinations or pseudohallucinations?]. AB - 'Hallucinations' were diagnosed in a 41-year-old Turkish woman after the death of her father. The nature of hallucinations is often difficult to understand in patients of Turkish (or Moroccan) origin, as they can be diagnosed as a psychotic feature, or as a sign of dissociation, reexperiences, mourning, personality disorder or depression. Most of the time the symptoms can be regarded as culture bound phenomena of psychosocial stress, often accompanied by somatoform complaints. In those cases they should be diagnosed as pseudohallucinations. Drug interventions only play a supportive role in the treatment of these symptoms. Joining the patients' explanatory model and exploring the psychological meaning of symptoms may be a rather difficult process which, however, may result in fewer symptoms in the end. PMID- 10812439 TI - [Panic disorder in patients with chest pain and palpitations: an often unrecognized relationship]. AB - The prevalence of panic disorder in patients who present with chest pain or palpitations to a First Heart Aid setting varies in the literature between 0% 59%. In a high percentage of cases, panic disorder is not recognized by the cardiologist in patients who present initially with chest pain or palpitations. Patients with panic disorder have a large and ongoing medical consumption. A selective serotonin reuptake inhibitor and/or cognitive therapy appear to be good treatment of panic disorder in patients who present initially with chest pain or palpitations. A CO2 challenge test elicits the symptoms in patients with panic disorder with high sensitivity and specificity but this test has not been validated in patients who present initially with chest pain or palpitations and in whom the diagnosis 'panic disorder' is not yet established. PMID- 10812440 TI - [Serum lactate level as a indicator of tissue hypoxia in severely ill patients]. AB - Adequate oxygen supply to the tissues is of vital importance to survive critical illness and trauma. Shock can be defined as an imbalance between oxygen demand and oxygen supply. Clinical features of shock, like hypotension, tachycardia, cold clammy skin et cetera, are poorly correlated with presence of tissue hypoxia. A high lactate level is an early sign of tissue hypoxia. In severely ill patients tissue hypoxia is the most important cause of increased lactate levels. Increased blood lactate levels are related to increased mortality. Optimizing oxygen supply by fluid resuscitation is the intervention of first choice. PMID- 10812441 TI - [Clinical thinking and decision making in practice. A student with sudden headache]. AB - A previously healthy man aged 18 years suddenly developed a severe headache, followed by nausea, vomiting, fever. During the following weeks a left-sided hemiparesis developed. CT of the brain revealed a pansinusitis and a frontal epidural empyema. A few days after surgical drainage his condition deteriorated and subsequent CT showed a right-sided subdural empyema. This was surgically evacuated and followed by long-term high-dose intravenous antibiotic therapy. The patient made a complete recovery. A sudden severe headache can be caused by an intracranial infection and intracranial pus collections can occur in the subdural as well as in the epidural space. Epidural empyema is a limited disease with relatively mild symptoms and a favourable prognosis, whereas subdural empyema may rapidly spread and cause severe disease with a poor prognosis. Quick antibiotic treatment and surgical drainage are required. PMID- 10812442 TI - [Panic disorder, chest pain and palpitations: a pilot study of a Dutch First Heart Aid]. AB - OBJECTIVE: To determine how many patients, presenting to a First Heart Aid (FHA) with chest pain or palpitations without a cardiac origin for their complaints, have a panic disorder and/or depression. DESIGN: Prospective and questionnaire investigation. METHOD: In 3 months (November 1st 1998-January 31st 1999) all patients presenting to the FHA of the University Hospital Maastricht, the Netherlands, and who were not admitted, were screened for the presence of psychopathology by means of a questionnaire, the 'Hospital anxiety and depression scale' (HADS). Patients scoring above 8 on the HADS with no cardiac cause for their initial complaint were interviewed using the 'Mini international neuropsychiatric interview' (MINI) to determine whether there was a panic disorder and/or a depressive episode. RESULTS: Of a total of 621 patients 251 met the inclusion criteria; 134 (53%) gave informed consent (72 (54%) men and 62 (46%) women, with a mean age of 55.9 (SD: 13.2; range: 23-84)). Of the 134, 77 had a HADS score > or = 8; in 59 (30 men; 29 women) the MINI was carried out; in 49 (83%) panic disorder (n = 45) or depression (n = 4) was diagnosed. In 7/45 the cardiologist had diagnosed a psychiatric disorder ('Hyperventilation'). CONCLUSION: In 83% of the patients who visited the Maastricht FHA with cardiac complaints but without a cardiac origin and who had a HADS score > or = 8, panic disorder and/or depression was diagnosed. PMID- 10812443 TI - [Pharmaco-economic evaluation of mandatory HIV-screening in pregnancy; a cost efficacy analysis in Amsterdam]. AB - OBJECTIVE: To estimate the cost effectiveness of universal screening for HIV of pregnant women in Amsterdam. DESIGN: Pharmaco-economic model calculation. METHOD: An estimate was made of the minimal and maximal prevalences of undiagnosed HIV infection during pregnancy for the whole of Amsterdam, based on epidemiological data from observation among pregnant women in two Amsterdam hospitals and one obstetrical practice. The calculation was based on universal screening with an ELISA test. The interventions after screening comprised pharmacotherapy during pregnancy, delivery by caesarean section and breast-milk substitution. The issues of pharmaco-economic analysis were whether or not costs were reduced and net costs per year of life gained; the question was also studied at what lifetime costs of care for HIV infected children the net costs would be nil (costs equal benefits). RESULTS: Universal HIV screening in Amsterdam required a total investment of about Dfl 300,000.-per annum. In many of the analysed options for HIV screening the financial profits exceeded the investment. Variation of assumptions showed that the net costs of HIV screening under all conditions investigated would remain below Dfl 1,200.-per life year gained. CONCLUSION: Universal HIV screening of pregnant women in Amsterdam showed a favourable cost effectiveness. The calculations indicated a possibility of reducing costs. PMID- 10812444 TI - [Acute renal insufficiency caused by bilateral ureteral obstruction after appendectomy in a 6-year old boy]. AB - A 6-year-old boy developed macroscopic haematuria on the 4th day after appendectomy for acute appendicitis, at which the appendix was found to be perforated. During the next few days the urine secretion decreased and malaise, pain in the lower abdomen, nausea and vomiting occurred. On a management of ample fluid administration, the urine secretion recovered and the symptoms subsided in a few days. In the early postoperative stage after appendectomy in children the possibility should be kept in mind of the development of acute renal insufficiency due to bilateral ureteral obstruction as a result of oedema of the posterior bladder wall, even if by means of ultrasonography only mild to moderate abnormalities are noted. Awaiting decompression by means of the introduction of bilateral ureteric stents, in order to prevent irreversible renal damage, supportive therapy with fluid administration depending on the diuresis seems indicated. PMID- 10812445 TI - [Clinical evaluation of efficacy and adverse effects in the (European) registration of drugs: what does it mean for the doctor and patient?]. AB - The clinical criteria for admission of new drugs to the European common market have become more stringent in recent years. Increasingly often, the manufacturer is required to demonstrate that the new drug offers a clinically visible and relevant benefit to the patient. Efficacy and adverse effects should not only be studied by comparative trials with placebo, the registration authorities also expect the drug to be compared with the standard treatment already available. Such trials should prove that the balance between efficacy and adverse effects of the drug is better than that of placebo and at least as good as the standard treatment, as regards not only statistical significance but also clinical relevance. Therefore, Dutch and European assessment reports and product information may be increasingly useful to prescribers, patients and insurers in determining the role and therapeutic value of new drugs within the existing therapeutic possibilities concerning certain diseases. PMID- 10812446 TI - [The miraculous growth of the 'quackery funds']. PMID- 10812447 TI - [In Process Citation] PMID- 10812448 TI - [The miraculous growth of the 'quackery funds']. PMID- 10812449 TI - [In Process Citation] PMID- 10812451 TI - [In Process Citation] PMID- 10812450 TI - [Miraculous growth of the 'quackery funds']. PMID- 10812452 TI - [Public health care after the Bijlmermeer airplane crash; the aftermath]. PMID- 10812453 TI - [Less medicalization of obstetrics in West Frisian islands than on the mainland]. PMID- 10812454 TI - [The operating room of the future]. PMID- 10812455 TI - Thyroid hormone metabolism and action in the brain and pituitary. AB - Brain and pituitary development, maturation, and function critically depend on thyroid hormone availability. The identification of several forms of nuclear T3 receptors, the region-specific expression of deiodinase isoenzymes in brain and pituitary, and the molecular analysis of thyroid hormone-responsive genes in fetal, newborn, and adult brain opened a new era in the understanding of thyroid hormone action. These integrated networks of receptors, deiodinases, and thyroid hormone responsive genes require strict regulation of thyroid hormone concentration at the right place and the appropriate time. Knockout and transgenic mouse models of components involved in hypothalamus-pituitary-thyroid periphery-feedback regulation revealed that lack of thyroid hormone (such as during iodine deficiency) leads to defects and phenotypes other than lack of thyroid receptor(s). In many aspects, expression of mutant thyroid receptors is worse than lack of wild-type receptors. Thyroid hormones control several genes in the CNS and are also essential for differentiation of pituitary lactotrophs and somatotrophs. Apart from most T3 effects which are mediated by nuclear receptors, T4 itself as well as its lower iodinated metabolites exert direct biological effects in the brain by mechanisms not involving nuclear T3-receptors. PMID- 10812456 TI - Interaction between psychotropic drugs and thyroid hormone metabolism--an overview. AB - Psychotropic drugs can influence synthesis and metabolism of thyroid hormones at different sites. Generally, lithium, tricyclic antidepressants and phenothiazines lead to a reduction in synthesis and/or metabolism of thyroid hormones. The induction of autoimmune thyroid disorders by lithium and phenothiazines has been proven in animal studies and possibly can also be found in humans. Antipsychotic drugs generally exert their therapeutic effects through a modulation of the monoaminergic and serotoninergic system. At the hypothalamic level, thyrotropin releasing hormone (TRH) is controlled by the monoamonergic system and by serotonin. Depending on the specific species, there is a particular and different influence on the secretion of different hypothalamic-pituitary-thyroid (HPT)-axis hormones. PMID- 10812458 TI - Genetic background of osteoporosis. AB - Osteoporosis is a systemic disorder of decreased skeletal mass as measured by bone mineral density (BMD), and disturbed skeletal architecture and function which results in an increased risk for bone fractures with consecutively increased morbidity and mortality. Twin and family studies have shown an important genetic component of BMD of about 40-60%. This exceeds other well known factors influencing BMD such as environmental factors like dietary calcium, physical activity or several drugs and diseases. Therefore, interest increased in the genetic background of bone mineral density. Polymorphisms of the Vitamin D receptor gene were the first to be published in this area. Studies on other loci or candidate genes such as the estrogen receptor gene or the collagen type I alpha1 gene also showed associations with bone mineral density that could explain at least a part of the genetic background of osteoporosis. Recently published data suggest that these genetic markers of bone metabolism are important in interaction with each other or in certain bone-affecting diseases. In the future, genetic studies on osteoporosis will have to screen further relevant genes and markers for bone metabolism as well as to evaluate the complex interactions of genetic influences, so that it would be possible to calculate a patient's individual risk for osteoporosis in the context of environmental influences. PMID- 10812457 TI - [Senile hypogonadism in man and hormone replacement therapy]. AB - Aging is accompanied by a progressive decline of testicular function. Whereas fertility persists until a very old age, endocrine function declines progressively and at age 70 yrs, more than 25% of men have hypogonadal testosterone levels. This decline in testosterone plays an important role in a series of signs and symptoms that accompany the aging process such as a decline in virility, in libido and sexual activity, muscle mass and strength, decline in bone mass (osteoporosis), an increase in abdominal fat mass and a decrease in the feeling of general well being. Most of these signs and symptoms have a multifactorial origin, nevertheless, androgen substitution generally improves most of these symptoms, increasing muscle mass and strength, improving libido and sexual activity, decreasing abdominal fat and improving insulin sensitivity as well as the sense of well being, effects which suggest a role of androgen deficiency in their genesis. Side effects of this substitutive therapy, mainly at the level of the prostate, with possible stimulation of a prostatic carcinoma, should be carefully looked for. Before starting any androgen therapy, the presence of a prostatic carcinoma should be excluded by rectal examination and PSA measurement, eventually, in case of doubt, complemented by transrectal echography. Presence of a prostatic carcinoma is an absolute contraindication for androgen therapy. During treatment, development of side effects should be traced by a six monthly rectal examination and PSA determination as well by monitoring of hematocrit and lipid profile. PMID- 10812459 TI - [Steroid-induced osteoporosis: pathogenesis and therapeutic consequences]. AB - Many chronic inflammatory diseases require treatment with steroids, however, a remarkable proportion of steroid-treated patients suffer from osteoporosis as major complication after longterm treatment. Steroid-induced osteoporosis represents one of the most important secondary causes of osteoporosis. The pathogenesis is complex, there exists evidence that steroids cause a reduction of circulating testosterone and estrogen concentration and adversely affect calcium balance. The most important mechanism is a decrease in osteoblastic activity. As a consequence loss of bone mineral density and increased risk of fracture develop. In spite of better understanding of the causal relationships preventive strategies were infrequently applied. If we care for patients with steroid therapy we have both to consider the problem of steroid-induced osteoporosis and to focus on strategies to evaluate patients at risk. The dosage of the steroid, life style factors (such as lack of exercise, alcohol consumption and smoking), menopausal status, low bone mineral density at baseline and previous osteoporotic fractures predispose for the manifestation of steroid-induced osteoporosis. Therapeutic decisions depend on risk factors of the individual patient. Supplementation of calcium and vitamin D is usually appropriate, and postmenopausal women should be offered hormone replacement therapy. The prescription of bisphosphonates is strongly recommended to patients at elevated risk. PMID- 10812460 TI - [Prevalence of diabetes in acromegaly and Cushing syndrome]. AB - In a retrospective study glucose metabolism was investigated in 206 patients with acromegaly and 131 patients with Cushing's disease. 40.5% of the patients with hypersomatotropism and 32.0% of the patients with hypercortisolism suffered from overt diabetes mellitus. Impaired glucose tolerance was present prior to therapy in 28.2% and 30.6% of the patients, respectively. In acromegaly the incidence of overt diabetes mellitus was higher in women than in men, but no difference existed in the distribution of impaired glucose tolerance between both sexes. No correlation was found between growth hormone levels and occurrence of diabetes. In acromegaly and Cushing's disease overt diabetes increased with advanced age. Diabetes mellitus occurred independently from the etiology of hypercortisolism. PMID- 10812461 TI - Scintigraphic and ultrasonographic appearance in different tumor stages of thyroid carcinoma. AB - RATIONALE AND OBJECTIVES: Scintigraphy is routinely used in evaluating thyroid nodules. Functioning nodules are reported to have a low probability of being malignant. Therefore cancer should appear hypo-functioning or "cold" on scintiscan. The aim of the study was to compare the scintigraphic pattern in different tumor stages of thyroid carcinoma. In addition, sonographic results are evaluated. In 151 patients with thyroid carcinoma 99mTc-pertechnetate scans were evaluated retrospectively by a visual inspection scoring method (A = no significant uptake to D = nodular uptake superior to normal thyroid tissue). Planar images were taken using a small field thyroid gamma camera. There were 52 patients with pT1 carcinoma (2 x follicular and 50 x papillary). The mean tumor size was 0.56 +/- 0.26 cm. The scintigraphic results were A and B in 5.7% (n = 6), C in 73% (n = 38), D in 15.6% (n = 8). Out of 40 patients with pT2 carcinoma, 34 had a papillary, 6 a follicular histology. Mean tumor size was 1.66 +/- 0.49 cm. The scintiscan was A in 12.5% (n = 5), B in 32.5% (n = 13), C in 42.5% (n = 17) and D in 12.5% (n = 5). There were 11 patients with pT3 carcinoma (4 x papillary, 7 x follicular). The mean tumor size was 3.96 +/- 0.88 cm in diameter. Scintiscan was A in 72.7% (n = 8), C in 27.3% (n = 3). Among 48 patients with pT4 carcinoma (2 x follicular, 1 x nondifferentiated, 45 x papillary), scan was A in 41.6% (n = 20), B in 14.5% (n = 7), C in 33.3% (n = 16) and D in 10.4% (n = 5). Mean tumor size was 2.16 +/- 1.45 cm (7 carcinomas < or = 1 cm, 23 x 1-2 cm, the remaining > 2 cm). Tumor size plays an important role in routinely used planar scintigraphy. Nodules greater than 2 cm in diameter tend to appear cold but microcarcinomas (< or = 1 cm) are often indifferent on scan. Therefore, planar 99mTc-pertechnetate scintigraphy is of little value in evaluating small thyroid nodules. In order to diagnose small thyroid nodules, ultrasonography and ultrasonographically guided FNAB should be recommended as the initial diagnostic steps in clinical routine. PMID- 10812462 TI - The role of F-18FDG PET in thyroid cancer. AB - Hypofunctioning nodules on scintiscan using Tc-99m Pertechnetate or I-123 have a higher probability of malignancy compared to eu- or hyperfunctioning nodules. However, in the preoperative assessment of thyroid nodules, ultrasonography and ultrasonography guided fine needle aspiration biopsy play the most important role, especially for papillary thyroid cancer. The problem of differentiating follicular adenoma from highly differentiated follicular carcinoma however remains. Also the additional use of a multi tracer imaging strategy (Tl-201/Tc 99m subtraction scan, Tc-99m Sestamibi, Tc-99m Tetrofosmin dual phase scintigraphy) has not solved this problem. Although it is unlikely, the question whether FDG PET is able to give a better differentiation between benign and malignant tumours in the preoperative assessment of thyroid nodules is not answered up to now. In contrast to preoperative diagnostics, FDG PET is of great value in the postoperative follow up of differentiated thyroid cancer. In case of elevated serum thyroglobulin but negative I-131 WBS FDG PET is the method of choice to detect I-131 negative recurrences and metastases. FDG uptake in metastases from differentiated thyroid cancer is correlated to low differentiation and maybe bad prognosis. There is also evidence that FDG PET may have a role in the follow up of anaplastic and especially in medullary thyroid cancer in the future. PMID- 10812463 TI - [Sonography of the thyroid gland in practice and in the clinic]. AB - Nowadays, the ultrasound examination is considered to be the most important procedure in the morphological examinations of the thyroid gland and its importance can be compared to the value of the basal thyroid stimulating hormone level (b-TSH) estimation in the examination of the thyroid function. Both methods in their preliminary stages can already exclude morphological and functional changes. In case of a disease, sonography makes the selection of further investigation procedures possible, in regard to their necessity and urgency. The organ size (in all 3 dimensions); the relationship to neighbouring structures; the echo pattern or the blood circulation are sonographic informations, which offer the specialist, the thyroid clinic or the surgeon (all with their wide differences of aims) an excellent means in the exclusion; the diagnosis; the investigation and the treatment strategy of thyroid gland diseases. PMID- 10812464 TI - [Change of surgical treatment in benign thyroid gland diseases-- influence of surgical management on perioperative complications]. AB - Since the beginning of 1994 standardized primary radical resection was performed in benign thyroid diseases at the surgical department, Barmherzige Brueder Hospital, Graz. In multinodular euthyroid goiter bilateral lobectomy or near total lobectomy respectively was done. In case of Graves' disease unilateral lobectomy and contralateral near total resection or "en bloc" thyroidectomy was performed. In accordance with literature a significant reduction of recurrent laryngeal nerve paralysis from approximately 9% to 1.2% occurred in our patients if this surgical procedure was performed consequently. Hypocalcemia rate after surgery needing treatment was approximately 1% at the time of demission. PMID- 10812465 TI - [Point mutation of thyrotropin receptors as the main cause of autonomously functioning thyroid nodules: experiences in the framework of our Styrian patients]. AB - Autonomously functioning thyroid nodules are frequently associated with mutations of the thyrotropin receptor. We analyzed a part of exon 10 of the thyrotropin receptor gene (base pairs 1762-1976) by direct sequencing and found missense mutations in 5 of 14 cases (codons 629, 631, 632, 633). Histologically, 3 of the 14 nodules were adenomas whereas 11 were hyperplasias. Nodules with mutations did not show significant differences from nodules without mutations with respect to age, histology, size, additional (non-functional) nodules and clinical symptomatology. Our results confirm that thyrotropin receptor mutations are involved in the development of autonomously functioning thyroid nodules. In this context, the terms hyperplasia and neoplasia should be reevaluated. PMID- 10812466 TI - [Seasonal variations in the diagnosis of hyperthyroidism]. AB - Seasonal variations in the frequency of thyrotoxicosis are known for a long time. Both, variations in the temperature and in the iodine supplementation are believed to affect the incidence of the diagnosis of hyperthyroidism. Seasonal variation of the incidence of hyperthyroidism was investigated before and after increase of salt iodination in 1990/1991. In the period from January 1987 to December 1995 hyperthyroidism was diagnosed for the first time on 4711 patients in our department. This population was subdivided according to etiology (Graves' disease, autonomous nodular goiter) and grade of hyperthyroidism (preclinical or clinical form). The so formed groups were divided according to the month of diagnosis into further 12 subgroups. Significant differences in month to month variation were found in the patients suffering from Graves' disease and autonomous nodular goiter with preclinical hyperthyroidism after increased iodine supplementation. The peak incidence of the diagnosis occurred in June, July, and August. Our results indicate that improved iodine supplementation may be the main cause of seasonal variations in the incidence of thyrotoxicosis. PMID- 10812467 TI - [Measurement of free triiodothyronine (FT3) using an electrochemiluminescence immunoassay in patients with autoantibodies to triiodothyronine]. AB - Thyroid hormone autoantibodies may lead to abnormal values of free triiodothyronine (FT3) and free thyroxine (FT4) by interference with the radio immunoassay (RIA). We examined thyroid function in six patients with known triiodothyronine-binding autoantibodies using a RIA and an electro chemiluminescence immunoassay (ECLIA). FT3 values measured by RIA were spuriously high, ECLIA measurement of FT3 led to correct values according to the patients' thyroid status. We conclude from these results that in patients with triiodothyronine-binding autoantibodies FT3 measurement by ECLIA is more useful than measurement by RIA. PMID- 10812468 TI - [Effect of aceclofenac on thyroid hormone binding and thyroid function]. AB - Influences of non-steroidal anti-inflammatory drugs (NSAID) on concentrations of thyroid hormones are known for a long time. These effects could be explained with interference between NSAIDs and thyroid hormone binding. We investigated the effects of a single dose of aceclofenac on thyroid function and thyroid hormone binding in 18 healthy volunteers. Serum levels of free thyroid hormones (FT3, FT4) and thyrotropin (TSH) were measured with commercial available kids and thyroid hormone binding was estimated with a specially modified horizontal argarose-gel-electrophoresis prior to and 2 hours after receiving a single dose of aceclofenac. We found a significant decrease in T3 binding on TBG and a significant increase of albumin-bound T3. All other investigated thyroid hormone binding parameters, FT3 and FT4, showed no significant changes. We conclude that aceclofenac leads to a significant redistribution of T3 protein binding. These effects seem to be explained by T3 displacement from TBG induced by aceclofenac. PMID- 10812469 TI - [Increased occurrence of autoimmune thyroiditis in patients with chronic rheumatoid arthritis]. AB - The frequent occurrence of both rheumatoid arthritis and autoimmune thyroiditis was already investigated with in part many conflicting results. We investigated a number of 792 patients (383 of them suffering from rheumatoid arthritis and 409 with osteoarthritis). In all patients antithyroid peroxidase and antithyroglobulin antibodies were determined. Patients with rheumatoid arthritis showed a significantly higher occurrence of circulating thyroid antibodies than those with osteoarthritis (9.1% versus 3.7%, p = 0.0016). We conclude that there exists a cumulate coincidence of both diseases. Patients suffering from rheumatoid arthritis should undergo a thyroid examination especially for the presence of autoimmune thyroiditis. PMID- 10812470 TI - The early days of bioinformatics publishing. AB - A brief history of the early days of publishing in the bioinformatics field is presented. PMID- 10812471 TI - Earliest pages of bioinformatics. AB - This review is a brief outline of the chronology and essence of early events in bioinformatics, covering the period from 1869 (discovery of DNA by Miescher) to 1980-1981 (beginning of massive sequencing). For the purpose of this review, bioinformatics is understood as a chapter of molecular biology dealing with the amino acid and nucleotide sequences and with the information they carry. PMID- 10812472 TI - A historical perspective on gene/protein functional assignment. AB - Sequence determination and analysis began on proteins in the 1950s, with RNA starting about a decade later and DNA a similar period later still. Hence many of the concepts for function prediction were first developed by looking at amino acid sequences. Over time these methods have become much more sophisticated, allowing better discrimination of only weak similarities. The most recent developments concern an examination of contextual information, such as operon structure, metabolic reconstruction or co-expression profiles. PMID- 10812473 TI - DNA binding sites: representation and discovery. AB - The purpose of this article is to provide a brief history of the development and application of computer algorithms for the analysis and prediction of DNA binding sites. This problem can be conveniently divided into two subproblems. The first is, given a collection of known binding sites, develop a representation of those sites that can be used to search new sequences and reliably predict where additional binding sites occur. The second is, given a set of sequences known to contain binding sites for a common factor, but not knowing where the sites are, discover the location of the sites in each sequence and a representation for the specificity of the protein. PMID- 10812474 TI - On the trail of protein sequences. PMID- 10812475 TI - History of the enzyme nomenclature system. AB - Naming things is essential for people to understand one another, no matter what language or field of interest is involved. This is as true for enzymes, genes and chemicals as it is for birds, food, flowers, etc. Effective communication requires a lack of ambiguity, but, in practice, ambiguities abound even between people who use the same language in different parts of the world, or even within the same country. Whereas ambiguities in the words used for common objects or actions have been the basis for many, more-or-less memorable jokes, they can also cause a great deal of confusion. Such linguistic chaos is welcomed by many as being a part of a diverse heritage that should be preserved at all costs to prevent us from descending into Orwellian 'newspeak'. However, in the sciences, there are distinct advantages in others being able to understand what one is doing. Many groups have stressed the need for standardized, universally accepted systems of nomenclature in chemistry, genetics, enzymology, etc. However, it is the universal acceptance that usually causes the problem. It is rare to find people who will admit that they find nomenclature to be an interesting subject, but many who profess contempt for it will get very excited if it is suggested that their pet nomenclature should be changed in the interest of clarity or uniformity. This account will consider the development of the enzyme nomenclature system, its benefits, shortcomings and future prospects. PMID- 10812476 TI - The early introduction of dynamic programming into computational biology. PMID- 10812477 TI - Serendipity in bioinformatics, the tribulations of a Swiss bioinformatician through exciting times! PMID- 10812478 TI - A brief history of genome research and bioinformatics in France. AB - The development of in silico genomics has progressed slowly in France for a number of political reasons. Two administrative organizations, the Groupement de Recherche sur les Genomes (GREG) and the Groupement de Recherche 1029 (GDR 1029) of the Centre National de la Recherche Scientifique (CNRS) have been established. These organizations have created the dynamics that hopefully will place France (which coordinated consortia that completed several of the first large microbial genomes) among the developed nations that support Large-Scale Biology. PMID- 10812479 TI - Do they have to take the eye out? PMID- 10812480 TI - Needle-stick injuries. PMID- 10812481 TI - Difference between ophthalmologists' and patients' perceptions of quality of life associated with age-related macular degeneration. AB - BACKGROUND: There may be a wide disparity between the perceptions of patients and those of their treating physicians concerning the quality of life associated with a given state of health. Because of this potential for difference of opinion, we performed a study to evaluate patients' and ophthalmologists' perceptions of quality of life, as measured by utility analysis, associated with visual loss secondary to age-related macular degeneration (AMD). METHODS: Cross-sectional study. Utilities were assessed, by means of both the time trade-off method and the standard gamble method, for various degrees of theoretical visual loss secondary to AMD for ophthalmologists-in-training and graduate ophthalmologists. These were compared to utilities for a known population of patients with actual visual loss due to AMD. A utility of 1.0 is associated with perfect health, whereas a utility of 0.0 is associated with death. RESULTS: With both the time trade-off and standard gamble methods, the patients had lower mean utilities than did the ophthalmologists for the same degrees of visual loss secondary to AMD. The ophthalmologists were significantly less willing than the patients to trade years of remaining life for perfect vision with the time tradeoff method (p < or = 0.01), and with the standard gamble method they were less willing than the patients to take the risk of dying in return for perfect vision. Given the scenario of counting fingers or worse vision in both eyes, the ophthalmologists were willing to trade 3.3 of every 10 years of remaining life for perfect vision in both eyes, whereas the patients with actual vision of counting fingers or worse in both eyes were willing to trade 6.0 of every 10 years of remaining life for this result. INTERPRETATION: When presented with the scenario of visual loss secondary to AMD, ophthalmologists substantially underestimated its effect on patients' quality of life. PMID- 10812482 TI - Polymerase chain reaction in the diagnosis of herpetic keratitis: experience in a developing country. AB - BACKGROUND: Herpetic ocular disease is a major cause of blindness. Rapid and accurate diagnosis is essential for prompt, proper treatment. We evaluated the usefulness of detection of herpes simplex virus (HSV) DNA by polymerase chain reaction (PCR) in the laboratory diagnosis of herpetic keratitis. METHODS: A retrospective study was conducted involving 234 patients who attended the cornea clinic at the Regional Ophthalmic Institute, Chennai, India, between March 1995 and September 1997. Inclusion in the study was based on clinical diagnosis of herpetic keratitis. Oligonucleotide primers directed against the HSV-I thymidine kinase gene were used, yielding a 110 base pair amplicon. The utility of PCR analysis was assessed against other diagnostic markers: HSV isolation on cell culture, HSV antigen detection by indirect immunofluorescence, detection of anti HSV IgG by enzyme-linked immunosorbent assay (ELISA) and detection of HSV specific tear secretory IgA (sIgA) by ELISA. These tests showed overall sensitivity values of 22.4%, 39.8%, 30.4% and 20.3% respectively. RESULTS: In epithelial keratitis all 35 specimens from which virus was cultured were positive by PCR. PCR gave a positive result in 23 (82.1%) of the 28 specimens in which HSV antigen was detected and in 4 (57.1%) of the 7 specimens that showed HSV-specific IgG. In addition, PCR detected HSV DNA in 5 of the 30 cases in which these three tests gave a negative result. PCR of two pooled tear samples (collected 1 week apart from the same patient) from 40 patients with stromal keratitis gave a positive result in 12 cases (30%). In stromal keratitis the sensitivity of PCR in detecting HSV DNA in tear samples was 85.7% with culture, indirect immunofluorescence and detection of anti-HSV IgG as the gold standard, and 80% with detection of sIgA as the gold standard. INTERPRETATION: The results confirm the good correlation with the clinical picture that can be obtained with PCR analysis. They also highlight the diagnostic utility of PCR in detecting HSV DNA in tear samples. This is particularly important in herpetic stromal keratitis, in which collection of corneal scrapings is not advised and, hence, conventional techniques such as virus isolation and antigen detection become difficult. PMID- 10812483 TI - The complication rate of edrophonium testing for suspected myasthenia gravis. AB - BACKGROUND: The incidence of life-threatening complications from edrophonium chloride (Tensilon) testing for suspected myasthenia gravis is thought to be extremely low. We carried out a survey to determine the rate of serious complications from such testing. METHODS: In April 1998, 357 physicians listed in the 1998 roster of the North American Neuro-ophthalmology Society were mailed a questionnaire for anonymous completion. Questions asked included the number of years the clinician had practised neuro-ophthalmology, the estimated number of edrophonium tests performed since completion of training, the number and nature of major complications from edrophonium, and whether the clinician preferred the sleep test or ice test to edrophonium testing. RESULTS: The response rate was 56% (199/357). Of the 199 respondents, 105 (53%) had practised neuro-ophthalmology for at least 10 years. The group estimated that they had performed at least 23,111 edrophonium tests, of which 37 (0.16%) were associated with a serious complication, mostly attributed to brady-arrythmias and syncope. Respiratory failure, seizure, severe vomiting and transient ischemic attack were also reported. Thirty-one respondents (16%) preferred the sleep test or ice test to the edrophonium test; one-third of this group reported a serious complication with edrophonium. INTERPRETATION: The rate of significant complications of edrophonium testing is low, but the complications can be potentially life threatening. Clinicians should know the nature and incidence of these complications when obtaining informed consent for edrophonium testing. PMID- 10812484 TI - Voluntary globe subluxation. PMID- 10812485 TI - Ultrasound biomicroscopy in diagnosis of a cyclodialysis cleft in a patient with corneal edema and hypotony after an air bag injury. PMID- 10812486 TI - Choroidal melanoma and rhegmatogenous retinal detachment. PMID- 10812487 TI - Palinopsia: a case report and review of the literature. PMID- 10812488 TI - Vaccination as an additional player in the mosaic of autoimmunity. PMID- 10812489 TI - Kidney transplantation is a safe therapeutic tool in systemic lupus erythematosus. PMID- 10812490 TI - Evaluation of gastrointestinal toxicity of ibuprofen using surrogate markers in rats: effect of formulation and route of administration. AB - OBJECTIVE: To elucidate the mechanism of gastrointestinal (GI) toxicity of ibuprofen and to examine the effect of altered site of drug release using gastroduodenal and intestinal permeability tests in the rat model. METHODS: Adult male Sprague-Dawley rats were administered (n = 6 per group) either: (1) 100 mg/kg immediate or sustained release ibuprofen; (2) 100 mg/kg immediate release and ibuprofen lysinate; or (3) 100 mg/kg or 200 mg/kg ibuprofen po or s.c. Upper and lower GI permeability as a surrogate marker of toxicity were determined at pre-determined times using the urinary excretion of orally administered sucrose and 51Cr-EDTA permeability probes, respectively. RESULTS: Ibuprofen administration resulted in a dose-dependent increase in both upper and lower permeability of the GI tract. Both immediate and sustained release preparations of ibuprofen increased upper and lower GI permeability with no shift of toxicity to the site of drug release. Ibuprofen lysinate also induced significant increased upper and lower GI permeability comparable to immediate release ibuprofen. Oral doses were not more toxic than s.c. doses. CONCLUSION: Ibuprofen induced increased GI permeable appears to be independent of the type of formulation and route of administration. This indicates that, contrary to some other nonsteroidal anti-inflammatory drugs, ibuprofen's effect on GI permeability is mainly systemic and the direct local effect contributes minimally to its overall GI toxicity. Ibuprofen may be a suitable candidate for sustained release formulations since its effect may be prolonged without the danger of a shift of side effect from the upper to the lower GI tract. PMID- 10812491 TI - Diffuse idiopathic skeletal hyperostosis (DISH): is it of vascular aetiology? AB - OBJECTIVE: Reassessment of the pathological features of spinal involvement in DISH and studying the possible aetiopathogenetic mechanism/s of DISH in view of clinical, radiological and pathological findings. METHODS: Forty Egyptian patients with DISH were included in this study. They underwent clinical and radiological assessment. Routine lab tests were done in addition to measuring blood sugar, serum lipids and uric acid. Pathologic study of 50 macerated specimens of fused spines fulfilling the criteria of DISH was also performed. A pathologic study of another 50 macerated specimens from normal spines were examined as a control. RESULTS: Radiological assessment showed spinal involvement in 100% of the patients in the lower thoracic region, while it was present in 75%, 70% and 55% in the upper thoracic, lumbar and cervical regions respectively. Pathological study revealed a significant increase in the number and width of nutrient foramina, denoting hypervascularity of the ossified ligaments and vertebrae involved (P < 0.001), in addition to a significant (P < 0.001) increase in the size of the affected vertebrae, pointing to the possible role of a vascular disorder in the disease pathogenesis. Metabolic disorders were evident among our group of patients in the form of obesity (50%), hyperlipidemia (80%), diabetes mellitus (60%), and hypertension (45%). CONCLUSION: DISH is a diffuse systemic condition which is most probably related to abnormal bone cell growth/activity reflecting the influence of metabolic factors that lead to new bone deposition. The vertebral blood supply is a predisposing factor that contributes to the onset/progression and/or localization of DISH. PMID- 10812492 TI - Broadening of the T cell receptor spectrum among rheumatoid arthritis synovial cell-lines in relation to disease duration. AB - OBJECTIVE: The aim of the study was to evaluate the T cell receptor (TCR) family usage in T cell-lines from subcutaneous nodules and synovium from patients with rheumatoid arthritis (RA), with specific reference to the duration of symptoms. In vitro adherence characteristics of nodular T cells was studied as well. METHODS: Monoclonal antibodies were used to determine the distribution of TCR families in T cell-lines from synovium of patients with early and long-standing RA, from rheumatoid nodules and control tissues. An in vitro binding assay with T cell-lines from 2 rheumatoid nodules was performed. RESULTS: In early RA synovium, a restricted TCR family usage was observed in 5 out of 8 patients, contrary to long-standing disease, peripheral blood, ileum and colon. In RA nodules, a similar degree of restriction was noted. Moreover, the same TCR family was overexpressed by T cell-lines from different nodules derived from the same patient. T cell-lines from rheumatoid nodules demonstrated a preferential in vitro adherence to rheumatoid synovium and rheumatoid nodules, while no binding was observed on skin or tonsil. CONCLUSION: The TCR spectrum among RA synovial cell-lines broadens in relation to the disease duration. The overexpression of the same TCR family in different rheumatoid nodules from the same patients, and the in vitro adherence of T cell-lines from rheumatoid nodules may be indicative for recirculation between the different disease manifestations in RA. PMID- 10812493 TI - Reduction in serum leptin and IGF-1 but preserved T-lymphocyte numbers and activation after a ketogenic diet in rheumatoid arthritis patients. AB - OBJECTIVE: To assess the clinical, immunological and hormonal effects of carbohydrate restriction in rheumatoid arthritis (RA) patients via the provision of a ketogenic diet. METHODS: Thirteen RA patients with active disease consumed a ketogenic diet for 7 days, providing the estimated requirements for energy and protein whilst restricting their carbohydrate intake to < 40 g/day. This was followed by a 2-week re-feeding period. Clinical and laboratory evaluations were carried out on days 0, 7 and 21. Changes in serum glucose, beta-hydroxybutyrate (beta-HB), leptin, insulin-like growth factor-1 (IGF-1) and cortisol were also measured at these time points. To study CD4+ and CD8+ lymphocyte responses, mitogen stimulated T-cell activation was assessed in heparinised whole blood via flow-cytometric analysis of CD69 expression. RESULTS: After the 7-day ketogenic diet, there were significant increases in serum beta-HB and cortisol, and significant decreases in body weight, the total lymphocyte count, serum leptin, IGF-1 and glucose. However, with the exception of morning stiffness, there were no significant changes in any of the clinical or laboratory measures of disease activity, or in early T-lymphocyte activation and the absolute numbers of CD4+ and CD8+ cells. CONCLUSION: In RA patients several of the metabolic and hormonal responses to a ketogenic diet, such as a fall in serum IGF-1 and leptin, resemble those which occur in response to acute starvation. However, the clinical and immunological changes which occur in response to acute starvation do not take place with a ketogenic diet and thus may be dependent upon energy and/or protein restriction. PMID- 10812494 TI - Dupuytren's disease in type I diabetic subjects: investigation of biochemical markers of type III and I collagen. AB - OBJECTIVE: To clarify whether biochemical markers of collagen type III and I metabolism show alterations in type I diabetic subjects with Dupuytren's disease (DD) compared to those without DD. METHODS: DD was assessed in a total of 28 type I diabetic subjects, mean age 43.4 +/- 9.5 (SD) and duration of diabetes 25.2 +/- 9.7 years. Concentrations of aminoterminal propeptide of type III procollagen (PIIINP), carboxyterminal propeptide of type I procollagen (PICP) and carboxyterminal cross-linked telopeptide of type I collagen (ICTP) in serum and excretion of cross-linked N-telopeptides of type I collagen (NTX) and deoxypyridinoline crosslinks (DPyr) into urine were measured. RESULTS: The prevalence of DD was 32% (9 of 28 diabetic subjects). Average serum ICTP was 2.7 +/- 0.8 micrograms/l in subjects without DD and 3.6 +/- 1.2 micrograms/l with DD (p = 0.0276). No significant association between other collagen markers and DD was found. The reference intervals of PIIINP and ICTP were exceeded only in 1 and 2 subjects, respectively, and they both had DD. CONCLUSION: The degradation of type I collagen might be increased in diabetic subjects with DD. The overall implication was that synthesis or degradation of type III and I collagen in diabetic subjects with DD did not differ enough from those without DD to reflect changes in the biochemical markers of type III and I collagen. PMID- 10812495 TI - Distribution of fibronectins and their integrin receptors in interface tissue from aseptic loosening of hip prostheses. AB - OBJECTIVES: To assess the distribution of fibronectins (FNs) and their integrin (Int) receptors in synovial membrane-like interface tissue (SMLIT) from aseptic loosening of total hip replacement (THR), and potential role of FN-Int interaction in the loosening process. METHODS: The alkaline phosphatase anti alkaline phosphatase (APAAP) method was used to detect the distribution of FNs and their Int receptors in SMLIT and control samples. Double immunofluorescence labeling was used to reveal the different co-localizations. RESULTS: Intensive FN staining appeared in the lining layers, sublining area, and vascular endothelium, while immunoreactivities for Int alpha 4, alpha 5, beta 1 subunits were detected in the lining and endothelial cells of SMLIT. Immunofluorescence labeling revealed Int alpha 5 and collagenase-1/collagenase-3 double positive cells in lining layers and sublining area of SMLIT. CONCLUSION: Increased expression of FNs, Int alpha 4 beta 1 and alpha 5 beta 1 appeared in SMLIT compared with that in OA synovial membrane. FN-Int interactions may play a role in local collagenase production. PMID- 10812496 TI - Anticardiolipin and antinuclear antibodies in cancer patients--a case control study. AB - OBJECTIVE: To investigate patients with cancer for the frequency of IgG and IgM aCL and ANA in comparison to a group of age- and sex-matched controls. METHODS: Serum levels of IgG and IgM anticardiolipin antibodies (aCL), antinuclear antibodies (ANA) and anticytoplasmic antibodies were evaluated in 145 cancer patients, including 20 patients with thromboembolic disease (TED) and compared with age- and sex-matched controls. RESULTS: Higher levels of IgG aCL were found in patients compared to controls (p < 0.02). However, there appeared to be no difference in serum aCL levels between TED and the remaining cancer patients. No difference was found in the frequency of antinuclear and anticytoplasmic antibodies between patients and controls and the autoantibody presence in patients was usually not associated with concomitant autoimmune disease. CONCLUSION: Apart from increased levels of non-thrombogenic associated IgG aCL, there was no evidence for significantly enhanced B cell autoreactivity in this large collection of cancer patients compared to controls. PMID- 10812497 TI - Aberrant proteolytic digestion of biglycan and decorin by saliva and exocrine gland lysates from the NOD mouse model for autoimmune exocrinopathy. AB - OBJECTIVE: The protein components of the extracellular matrix (ECM) are responsible for driving tissue morphogenesis, the development of differentiated function, and the sequestration of biologically active molecules such as growth factors in close proximity to tissue and organ cells. Recent reports indicate that saliva and exocrine tissue lysates from Sjogren's syndrome patients and the non-obese diabetic (NOD) mouse model for autoimmune exocrinopathy demonstrate elevated levels of specific enzymes that degrade the ECM, especially the matrix metalloproteinases (MMPs). To determine if elevated levels of MMPs could be important in exocrine tissue destruction, we examined proteolytic activity against two ECM proteoglycans, decorin and biglycan. METHODS: Purified recombinant human core protein for decorin or biglycan was incubated with saliva or gland lysates from either control BALB/c or NOD mice. Degraded proteoglycan products were estimated by Western blotting analysis using anti-decorin or anti biglycan monospecific polyclonal antibodies. The levels of TGF beta protein were measured by ELISA. RESULTS: Proteolytic activity for decorin and biglycan was not observed in the saliva and salivary gland lysates collected from C57BL/6 or BALB/c mice used as normal controls. In contrast, both proteoglycans were degraded by saliva and exocrine gland lysates from NOD mice and the congenic partner strains NOD-scid and NOD.B10.H2b. This proteolytic activity for proteoglycans was inhibited by the MMP inhibitors, EDTA, GM6001 and 1,10 phenanthroline. Protein steady state levels for TGF beta were increased in the saliva and gland lysates from 20-week old NOD strains, as compared to BALB/c mice and NOD treated with the MMP inhibitor GM6001. With the inhibition of MMP activity, TGF beta levels declined in saliva and gland lysates. CONCLUSION: Proteolytic degradation of the ECM molecules decorin and biglycan is elevated in the exocrine tissues of the NOD mouse model for Sjogren's syndrome. Furthermore, the proteolysis of small leucine-rich proteoglycans correlates with the presence of elevated levels of TGF beta in gland lysates and saliva. PMID- 10812498 TI - Features of polymyositis and dermatomyositis in the elderly: a case-control study. AB - OBJECTIVE: Polymyositis (PM) and dermatomyositis (DM) are uncommon idiopathic inflammatory myopathies (IIM). Little is known about these diseases in the elderly. We attempted to define the characteristics of PM/DM in the elderly by a case-control study involving the retrospective review of medical files of PM/DM patients. METHODS: We drew from among 200 PM/DM patients being followed in our Internal Medicine Department 21 patients (14 F/7 M), aged > or = 65 years at the onset of myositis (17 PM/4 DM) (mean: 69.9 +/- 4.8 yrs.). They were compared with 21 (15 F/6 M) randomly selected younger patients with IIM: PM (14) and DM (7) (mean: 46.4 +/- 12.4 yrs). Clinical, biological, electrophysiological and pathologic features, treatment regimens and side-effects in the 2 groups were collected. RESULTS: Clinical features were similar for the 2 groups. Elderly patients tended to have a higher frequency of cancer (24% vs 9.5%, p = 0.06), particularly of rectal adenocarcinoma. The time from disease onset to diagnosis was significantly longer in older patients (26 +/- 37 months vs 9 +/- 15 months; p = 0.02), normal CK levels were more frequent (40% vs 5%; p = 0.02) and serum CK levels were lower than for the population as the whole (11.5 N vs 22 N, p < 0.03). Electromyography features were more frequently suggestive of a chronic form of the disease in elderly patients. Treatment regimens and short-term side effects were similar for the 2 groups. CONCLUSION: PM and DM are often diagnosed late in the elderly. Biological data and electromyography features argue for a chronic form of the disease in this age group. Clinical and endoscopic rectal examinations should be carried out in elderly patients with PM/DM. PMID- 10812499 TI - High toxicity of sulfasalazine in adult-onset Still's disease. AB - OBJECTIVE: Sulfasalazine (SSZ) is an anti-rheumatic drug that has been used to treat chronic arthritis. In many reports, the use of SSZ in children with systemic onset juvenile rheumatoid arthritis (JRA) revealed frequent side effects which required discontinuation of the drug. We examined whether there were frequent side effects of SSZ in patients with adult-onset Still's Disease (AOSD). METHODS: From July 1990 to April 1998, we followed 41 AOSD patients. Ten were given SSZ for the treatment of arthritis and the side effects were studied. We also studied 109 consecutive patients with RA who had been given SSZ, as a control group. In addition, we retrospectively studied the side effects and efficacy of SSZ in both groups through their medical records. RESULTS: Six patients (60%, p < 0.01) with AOSD experienced side effects ranging from mild ones like abdominal pain, nausea and vomiting, urticaria, and facial flushing to severe ones such as high fever, hypotension, and severe myelosuppression as well as fulminant hepatitis, which led to the death of one patient. However, 16 patients (14.7%) with RA stopped using SSZ due to mild side effects such as rash, urticaria, gastrointestinal troubles, mild leukopenia, and fever. Three AOSD patients (30%, p = 0.053) and 15 RA patients (13.8%) stopped using SSZ due to its inefficacy. CONCLUSION: We conclude that SSZ appears to have frequent severe side effects in AOSD, as in systemic onset JRA. These potential adverse effects of SSZ should be considered when it is used to treat chronic arthritides with systemic symptoms. Further study of SSZ in the treatment of AOSD in a multi-center, placebo-controlled environment is needed. PMID- 10812500 TI - IL-8 producing cells in patients with Behcet's disease. AB - OBJECTIVE: IL-8 is thought to be the principal chemokine responsible for neutrophil activation and tissue infiltration in patients with Behcet's disease (BD). In various studies serum levels of IL-8 were reported to be increased. METHODS: IL-8 mRNA was purified from both peripheral whole blood samples and separated lymphocytes, granulocytes and monocytes of patients with BD and compared to that from healthy (HC) and disease controls. IL-8 sequences were revealed by PCR amplification using appropriate sequence-specific primers. mRNA levels were determined semi-quantitatively using an image analyser in comparison with beta-actin. RESULTS: Although the differences did not reach statistical significance, BD patients tended to have higher IL-8 mRNA levels compared to HC in whole blood samples (2.0 +/- 1.4 vs 1.5 +/- 1.2) as well as in their lymphocytes (2.7 +/- 1.6 vs 1.5 +/- 0.9). No differences were observed between BD and HC in monocyte or granulocyte IL-8 mRNA levels. CONCLUSION: Our results suggest that the cellular source of IL-8 is diverse in BD with a possible major contribution by lymphocytes. PMID- 10812501 TI - Catastrophic antiphospholipid syndrome associated with crescentic glomerulonephritis: a clinicopathologic case. AB - The association of renal failure with catastrophic antiphospholipid syndrome has been reported in the context of microvascular occlusions and/or malignant hypertension. We describe a 36-year-old woman who died of multiorgan failure with the laboratory, clinical and histopathological characteristics of catastrophic antiphospholipid syndrome associated with a crescentic glomerulonephritis and renal failure. PMID- 10812502 TI - Arthritis following combined vaccine against diphtheria, polyomyelitis, and tetanus toxoid. AB - We describe 2 cases of arthritis following immunization against diphtheria, poliomyelitis and tetanus toxoid. One patient developed monoarthritis of the knee after immunization, that regressed following synovectomy. Five years later, the arthritis recurred after a booster vaccine injection. One day after immunization, another patient developed arthritis of the ankle that persisted for 3 days. It is difficult to know whether there is a coincidental or a causal relation between immunization and arthritis. Although our cases suggest that immunization against diphtheria, poliomyelitis and tetanus toxoid may cause arthritis, additional cases must be reported before studies aimed at confirming this possibility are considered. PMID- 10812503 TI - The induction of experimental vascular diseases by immunization with pathogenic autoantibodies. PMID- 10812504 TI - Hypertrophic osteoarthropathy after cerebrovascular insult. PMID- 10812505 TI - Analgesic-antiinflammatory effect of a 100 Hz variable magnetic field in RA. PMID- 10812506 TI - Three cases of Buerger's disease associated with hyperhomocysteinemia. PMID- 10812507 TI - Septic arthritis caused by Stenotrophomonas maltophilia in a patient with acquired immunodeficiency syndrome. PMID- 10812508 TI - Endocrine disorders and antiphospholipid antibodies. PMID- 10812509 TI - Multiple cerebral abscesses in a woman with giant cell arteritis. PMID- 10812510 TI - Juvenile arthritis in Turner's syndrome. PMID- 10812511 TI - Behcet's disease with a cecal perforation. PMID- 10812512 TI - Remitting seronegative symmetrical synovitis with pitting edema and primary Sjogren's syndrome. PMID- 10812513 TI - Joint hypermobility syndrome and bilateral total occlusion of the ulnar arteries presenting as Raynaud's phenomenon. PMID- 10812514 TI - Spondylodiscitis due to Staphylococcus lugdunensis. PMID- 10812515 TI - Peculiar myelopathy in a patient with overlap syndrome with lupus- and rheumatoid like symptoms. PMID- 10812516 TI - Overview of lasers in plastic surgery. AB - The laser used today in plastic surgery has gone through many stages of development. This article examines the history of lasers, including the scientific background and the theory of selective photothermolysis in therapy. In addition, current trends and future developments in laser treatment are discussed. PMID- 10812518 TI - Laser tattoo removal. AB - The availability of Q-switched ruby Nd:YAG and alexandrite lasers has revolutionized the treatment of tattoos. These modalities offer significant advantages over all previously available treatments and are currently the standard of care for the cosmetic removal of unwanted tattoos. PMID- 10812517 TI - Laser treatment of vascular lesions. AB - The principle of selective photothermolysis, which states that thermal destruction can be predicted by choosing the appropriate wavelength, pulse duration, and pulse energy for a particular target, applies to the laser treatment of vascular lesions. The most commonly used vascular lesion lasers and light sources emit wavelengths of 532 nm, 577-600 nm, and 1064 nm. A wide range of vascular lesions can be treated using these systems. PMID- 10812519 TI - Laser treatment of pigmented lesions. AB - Lasers are powerful tools in the treatment of benign pigmented lesions. Their role in the treatment of pigmented lesions other than those which are benign has not been defined. As with other ablative treatments, avoidance of lesions with malignant potential is essential. The wise adage First do no harm applies. By presenting the patient with a realistic goal and expectations, as well as discussing alternatives, limitations and risks, an informed decision for laser treatment can be made. Care and conservatism protect both the patient and the integrity of the laser surgeon. PMID- 10812520 TI - Laser hair removal. AB - We are in a state of evolution in long-term hair removal. Safe and effective technology is now available. Lasers will always become smaller, faster, and perhaps more effective. For those interested in offering this procedure, evidence indicates that we are beyond the experimental phase. The treating physician should carefully review current available technology and receive proper training specific to that system. PMID- 10812521 TI - Laser incisional surgery. AB - Soft tissue surgery is more efficient, and sometimes more effective, using laser incisional surgery as opposed to standard techniques. The modern generation of lasers produces soft tissue incisions with a minimum amount of thermal damage at wound edges. The transition to laser incisional surgery requires a knowledge of methods, maneuvers, and safety information. PMID- 10812522 TI - Laser resurfacing for dermal photoaging. AB - The skin is a highly organized system composed of resident cells, extracellular matrix, blood vessels, and circulating cells that all work together to maintain cutaneous integrity. Environmental insults, particularly sunlight, act to alter the skin permanently, producing visibly undesirable effects. By wounding the skin or inducing a healing response with minimal wounding, the repair process can be activated to return the skin to a more normal condition. Owing to the complexity of the healing response, even the most well-studied and precise laser system can result in unpredicted results when used to treat photo-damaged skin. Through continued research into the normal functioning of skin, the alterations brought about by chronic photodamage, and the repair process, an integrated approach to treatment of photoaging will evolve. Agents such as alpha-hydroxy acids, retinoids, and growth factors that impact the healing response can be combined with various lasers to optimize improvement of photo-damaged skin, while minimizing the adverse consequences of treatment. PMID- 10812523 TI - Carbon dioxide and ER:YAG laser resurfacing. Results. AB - Laser resurfacing is exciting "futuristic" surgery. The CO2 laser resurfaces using different parameters from the Er:YAG laser. When the surgeon understands these parameters, each laser can be used as a powerful tool for specific clinical applications. The Er:YAG laser was initially thought to be for the patient who has minimal skin laxity, but who desires skin resurfacing and needs a speedy return to social life. The CO2 laser has typically been thought to work best for skin laxity as well as rhytids, at the price of a longer recovery period. As the hardware and techniques continue to evolve, the differences between the clinical scope addressed by each laser diminishes. Both lasers deserve a place in the plastic surgeon's armamentarium. This new combination CO2/Er:YAG technique is intriguing and deserves further in-depth investigation. Laser resurfacing is not a cureall, but, when applied appropriately, it is an excellent tool that the plastic surgeon can use for skin rejuvenation. PMID- 10812524 TI - Postoperative laser care. AB - Postoperative laser care is critical to the successful performance of CO2 and erbium laser resurfacing. Anticipation and prevention of postoperative problems and complications leads to greater patient satisfaction and fewer permanent sequalae. In most situations, a team approach, with the use of trained nurses and aestheticians, ensures greater success in this critical postoperative period. Laser resurfacing is one of the few procedures in which the surgery is only beginning when the surgeon leaves the operating suite. PMID- 10812525 TI - Variable-pulse width ER:YAG laser resurfacing. AB - New resurfacing laser systems have been introduced that offer the ablative capacity of Er:YAG lasers with the ability to add controlled degrees of thermal effect to provide collagen tightening and hemostasis. This article discusses the current systems available and offers comparisons to short-pulse Er:YAG and carbon dioxide lasers. A histologic study and initial clinical results are presented and discussed. PMID- 10812526 TI - Simultaneously combined ER:YAG and carbon dioxide laser (derma K) for skin resurfacing. AB - The simultaneously combined Er:YAG and carbon dioxide laser is ideally suited for treating deeper wrinkles with greater accuracy than either laser alone. By combining the precise ablative properties of the Er:YAG laser with the coagulative properties of the carbon dioxide laser, it is possible to control the depth of skin resurfacing to minimize complications and improve difficult and substantial wrinkles. PMID- 10812527 TI - Nonablative resurfacing. AB - Nonablative, or subsurface remodeling, is the newest approach to improving photodamaged skin. Because the degree of collagen remodeling is not as great as that of other, more destructive ablative approaches, the process can be ideal for individuals with limited time, wishing to improve (with lasers) the quality of their sun-damaged skin. In addition, nonablative laser systems may be used to maintain the rejuvenating effect of ablative laser systems. PMID- 10812528 TI - Lasers, facelifting, and the future. PMID- 10812529 TI - Dose of selective serotonin uptake inhibitors across pregnancy: clinical implications. AB - The use of antidepressants during pregnancy has undergone considerable scrutiny with respect to safety issues, though limited data with respect to dose management and symptom resolution is available. Previous reports on tricyclic antidepressants (TCAs) have demonstrated the need to adjust maternal dose later in pregnancy to maintain therapeutic serum concentrations. However, there is no data on the dosage of selective serotonin uptake inhibitors (SSRIs) required to maintain symptom resolution in women treated for major depression during pregnancy. The purpose of this study, then, was to assess the medication dosage requirements of SSRIs during this time. In this naturalistic study, pregnant women with a primary diagnosis of major depression were followed prospectively through pregnancy at monthly intervals with symptom assessment. Subjects were included in data analysis if they presented prior to 28 weeks gestation, were treated with SSRI monotherapy, received all psychiatric treatment during the pregnancy at the Emory Pregnancy and Postpartum Mood Disorders Program, and achieved euthymia after initial treatment intervention (CGI = 1 and Beck Depression Inventory (BDI) < 9) during pregnancy or failed to respond after eight weeks of treatment. Medication selection was based on personal treatment history or family treatment history (if any), and the published data on SSRIs in pregnancy. All medication dose adjustments were based on depressive symptoms as measured by the BDI and a psychiatric interview (ZNS). Thirty-four pregnant women were included in final analysis. Two thirds of the subjects (n = 22) required an increase in their daily dose of medication to maintain euthymia. The dose increases occurred at 27.1 +/- 7.1 weeks gestation, with mean BDI scores of 16.4 +/- 9.6, compared to a mean treatment response BDI of 6.9 +/- 5.4. Subject's age, education, past personal and familial psychiatric history were not significantly associated with dose adjustment. These novel data on SSRI daily dose in pregnancy parallels the extant literature with tricyclic antidepressants (TCA). Further work to determine the predictors of dose adjustments will provide guidelines for minimizing fetal exposure to both medication and maternal mental illness. PMID- 10812530 TI - Comparison of pramipexole, fluoxetine, and placebo in patients with major depression. AB - Pramipexole, a dopamine D2 receptor agonist, was tested in 174 patients with major depression, with or without melancholia and without psychotic features. Three daily dose levels (0.375 mg, 1.0 mg, and 5.0 mg) were compared to fluoxetine (Prozac) at 20 mg and placebo in a randomized, double-blind, parallel group study. After a 1 week placebo run-in period, patients were treated for 8 weeks, had a post-study follow-up (week 9), and were evaluated primarily with the Hamilton Psychiatric Rating Scale for Depression (HAM-D), the Montgomery-Asberg Depression Rating Scale (MADRS), and the Clinician's Global Impressions-Severity of Illness scale (CGI-SI). All patients who received one dose of study medication were included in the observed-case analysis (no missing data were replaced). Results indicated that by endpoint (week 8), patients receiving pramipexole at the 1.0 mg per day dose had significant improvement over baseline compared to the placebo group by measure of the HAM-D, MADRS, and CGI-SI. Significant improvement in this dose group was seen at other timepoints as well. The most obvious improvement was seen in the pramipexole 5.0 mg group, although a substantial dropout rate for this group precluded statistical tests vs. placebo late in the study. Patients taking fluoxetine also showed significant improvements at endpoint on the MADRS and earlier in the study on the HAM-D. No new or unusual safety concerns were generated during this study. Pramipexole helped safely alleviate the symptoms of depression at 1.0 mg per day and especially in those patients who could tolerate the escalation to 5 mg per day. PMID- 10812531 TI - Postpartum and nonpostpartum depression: differences in presentation and response to pharmacologic treatment. AB - Following childbirth, major depression (postpartum depression) affects approximately 8-12% of new mothers. However, little is known about the pharmacological management of postpartum depression, and no studies to date have assessed differences in treatment response between women with postpartum and nonpostpartum major depression. The authors reviewed the records of 26 women with postpartum major depression and 25 women with major depression unrelated to childbearing (nonpostpartum depression) who presented to them for treatment over a 4-year period. Compared with the nonpostpartum depressed patients, the postpartum depressed women were significantly more likely to present with anxious features. Also, cases of postpartum depression were more severe than cases of nonpostpartum depression. While the postpartum patients were equally as likely to recover (as defined by a Clinical Global Impression score of 1 or 2) compared to the nonpostpartum-depressed patients, their time to response was significantly longer. By 3 weeks of pharmacotherapy, 75% of the nonpostpartum cases had recovered, in contrast to only 36% of the postpartum cases. Further, postpartum patients were significantly more likely to be receiving more than one antidepressant agent at the time of response to treatment. Length of depression prior to treatment did not explain the difference in treatment response. Presence of depressive symptoms during pregnancy and timing of onset of the depression (before vs. after 4 weeks of delivery) did not affect likelihood of treatment response in this sample. Women with postpartum depression appear to be significantly more likely than the nonpostpartum women to present with anxious features, take longer to respond to pharmacotherapy for depression, and require more antidepressant agents at the time of response to treatment. PMID- 10812532 TI - Melancholic features and response to lithium prophylaxis in mood disorders. AB - Clinical predictors of the efficacy of lithium prophylaxis in mood disorders have great potential value. Melancholic features during depressive phases have been both proposed and rejected as valid predictors of favorable outcome. The aim of the present study is to describe the validity of melancholic features during depressive phases as predictors of the prophylactic efficacy of lithium. Sixty one subjects affected by bipolar (n = 51) and major depressive (n = 10) disorder were followed prospectively for an average of 53 months. All subjects were evaluated as a lifetime perspective at intake, by the Operational Criteria checklist for psychotic illness (OPCRIT). Melancholic features were correlated with outcome only when controlling for time of first lithium administration. These two variables accounted for more than 30% of the total variance in lithium response. Others clinical factors such as polarity, delusions, gender, onset, personality disorders, and family history of mood disorders did not influence the observed association. Our preliminary findings suggest that melancholic features may be associated with favorable lithium prophylactic outcome in mood disorders. PMID- 10812533 TI - Treatment of depressive mood in schizophrenia with the atypical antipsychotic quetiapine. AB - Two patients with schizophrenia and depressive mood experienced remission in both their psychotic and depressive symptoms during treatment with the atypical antipsychotic quetiapine. These case reports illustrate the antipsychotic clinical efficacy of quetiapine and its antidepressant effects in the treatment of patients with schizophrenia and depressive mood. PMID- 10812534 TI - Correlation between temperature and vibration thresholds and somatosensory evoked potentials. AB - The psychophysically assessed thermal specific, thermal pain and vibration sensitivities were correlated to somatosensory evoked potentials in eighteen patients with definite multiple sclerosis. In the psychophysical tests, modality specific stimuli were used. Somatosensory potentials were electrically evoked. The abnormalities of both the temperature and the vibration sensitivity were to same extent related to the somatosensory evoked potentials. Dorsal columns-medial lemnisc and anterolateral-spinothalamic demyelinating lesions were presumed. The psychophysical tests supplement the clinical, laboratory, neuroradiologic and electrophysiological tests. These should be included in the battery of diagnostic tests in multiple sclerosis. PMID- 10812535 TI - Nerve conduction study of the medial and lateral plantar nerves. AB - The medial and lateral plantar nerves may be evaluated through the recordings of the compound sensory nerve action potentials (CSNAP), compound mixed nerve action potentials (CMNAP) and compound muscular action potentials (CMAP). As some of these potentials are not easily and always obtainable in normal individuals, our purpose was to verify the consistency of these potentials for the study of these nerves. Fifty-one normal adult volunteers were examined. The CSNAP, CMNAP and CMAP, related to the medial and lateral plantar nerves were evaluated bilaterally. CSNAP were not obtained in 7.8% and in 17.6% from the medial and lateral plantar nerves respectively. CMNAP from the lateral plantar nerve were not obtained in 15.6%. CMNAP from the medial plantar nerves and CMAPs from the abductor hallucis and abductor digiti quinti were obtained for all nerves tested. Our results, therefore, suggest that these last 3 parameters are the ones more reliable for clinical application. PMID- 10812536 TI - Electromyography of the infrahyoid muscles: pathological and normal findings. AB - In recent years, the infrahyoid muscles (IHM) have been used by plastic reconstructive surgeons as a neurovascular muscle flap in the neck and mouth region. METHODS: A preoperative electromyographic examination (EMG) of the IHM was performed in 10 patients, of whom 9 suffered from tongue cancer, in order to detect neurogenic lesions caused by possible metastases or lymph nodes. The results were compared to those of 10 healthy controls. RESULTS: The EMG at rest showed no pathological spontaneous activity in any patient. During light voluntary innervation, the motor unit potentials (MUPs) were normal in controls and in patients with normal sonographic images, computertomographic scans, and histologic findings after surgery in the neck region. When metastatic lymph nodes were found on one side of the neck, the number of polyphasic MUPs in the IHM of that side was increased in some cases (n = 6), and normal in others (n = 5). Traumatic or radiogenic lesions clearly resulted in pathological EMG findings (n = 6). A maximal innervation of the IHM was achieved during head bending and jaw opening, there was no activation of the IHM with tongue movements and vice versa. CONCLUSIONS: If the presence of lymph node pathology was demonstrated using imaging techniques, a resulting lesion of the ansa cervicalis can functionally be demonstrated by EMG. In patients without lymph node metastases and without concurrent other lesions in the cervical region, EMG of the IHM seems to give no further clinical information. A clear postoperative functional differentiation of the transplanted IHM and the indigenous tongue muscles is possible. PMID- 10812537 TI - Differential diagnosis of amyotrophic lateral sclerosis and similar syndromes. AB - The aim of this investigation was to re-evaluate the electromyographic findings in patients with ALS and with localised lesions of the spinal cord and to find some clues for differentiation. Two hundred and sixteen patients took part in this investigation. They were subdivided into two groups. The first group consisted of 168 patients with data for amyotrophic lateral sclerosis. The second group consisted of 48 patients with syndrome of amyotrophic lateral sclerosis caused by cervical myelopathy. Detailed neurological and electromyographic examinations were performed to all patients. In conclusion the electromyographic findings in patients with ALS are not different than the abnormalities in muscles innervated by the affected segments in patients with localised lesions which compress the anterior portion of the spinal cord. The only clues for differentiation are the clinical signs and the distribution of electromyographic abnormalities. PMID- 10812538 TI - Muscle fiber conduction velocity during isometric contraction and the recovery period. AB - Changes of muscle fiber conduction velocity (MFCV) detected by surface array electrodes during an isometric contraction and the recovery period were evaluated. The location on skin measured for action potentials of muscle fiber in m. biceps brachii was a distance of 5 mm and 30 mm from the end-plate to the distal tendon. The MFCV was evaluated by averaging raw EMG waves. The MFCVs at both locations declined gradually during the loads of sustained isometric contractions of 30, 50, and 70% of the maximum voluntary contraction (MVC). The degree of the decrease of the MFCV was extremely intense during a sustained contraction of 70% MVC. The values of the MFCV at the location of 5 mm from the end-plate in the period close to the exhaustion state showed a significant decrease compared with the values in the initial period during the contractions for the above three kinds of loads, while the decrease of the MFCV at the location of 30 mm from the end-plate was not found to be so significant during the contractions. In the recovery period, the contraction of 5% MVC was maintained, and the resultant MFCVs restored gradually to the value of the initial period. At the location of 5 mm from the end-plate, the MFCVs at 15 minutes after the end of the load were significantly higher than those just after the end of the load. At the location of 30 mm from the end-plate, the increases of the MFCVs during the recovery period did not show significant changes. Changes of the MFCV during the isometric contraction and the recovery period depend greatly on the location of the electrodes measured for the action potentials of the muscle fibers. PMID- 10812539 TI - Effect of muscular ultrasound stimulation on power spectrum electromyography during a strengthening training. AB - The purpose of this study is to describe the electromyographic (EMG) power spectral evolution during a muscle strengthening training based on ultrasound stimulation applied to the right rectus femoris muscle simultaneously with its maximum voluntary isometric contraction (MVIC) in ten healthy subjects. From the surface EMG of rectus femoris the Zero Crossing (ZC) and integrated EMG (IEMG) were calculated. The Mean Power Frequency (MPF) and Median Frequency (MF) from the EMG power spectrum were obtained using a maximum entropy method. An Analysis of Variance (ANOVA) for repeated measures showed significant changes for IEMG and ZC throughout sessions and for MPF, MF and ZC between the pretest and posttest. Immediate increases in MPF, MF and ZC were observed (paired t-test, p < 0.05) after training and an increment of IEMG during sessions (one-way ANOVA, p = 0.01). The ultrasound pulsed stimulation applied simultaneously with a MVIC produced the development of the muscle force. The mechanism of action of this application could be the mechanical effect, compression and rarefaction, produced by the ultrasound stimulation during the muscle contraction process. PMID- 10812540 TI - The biophysical effects of ultrasound on median nerve distal latencies. AB - PURPOSE: Previous studies have documented the lack of ultrasound's non-thermal effects on nerve conduction using frequencies of 1 MHz and 870 kHz. The analyses and conclusions were reached, despite only one study incorporating pulsed ultrasound. The purpose of this study was to determine the biophysical effects of continuous wave (CW) and pulsed wave (PW) ultrasound on median nerve motor and sensory latencies using common frequencies of 1.0 and 3.0 MHz. SUBJECTS: Fifteen healthy subjects (8 males, 7 females, age = 23.5 + 4.44 yrs, height = 171.2 + 10.7 cm, weight = 67.5 + 7.9 kg) without a history of neurological or musculoskeletal injury to their non-dominant arm volunteered for testing. METHODS AND MATERIALS: Subjects were exposed in counterbalanced order to five ultrasound treatment conditions: (1) 1 MHz, 1.0 W/cm2, 8 min., (2) 1 MHz, 1.0 W/cm2, 50% PW, 8 min., (3) 3.0 MHz, 1.0 W/cm2, CW, 8 min., (4) 3.0 MHz, 1.0 W/cm2, 50% PW, 8 min., (5) placebo, 0.0 W/cm2, 8 min. Dependent measures for motor and sensory latencies, and subcutaneous temperatures were taken pretreatment, at 2, 4 and 6 minutes during treatment, and immediately post-treatment. Separate two within repeated measures ANOVA were used for each dependent measure. RESULTS: Analysis revealed significant interactions for motor latencies [F (16,224) = 52.77, p < .001], sensory latencies [F (16,224) = 41.10, p < .001], and subcutaneous temperatures [F (16,224) = 52.77, p < .001]. Tukey's HSD post hoc analyses confirmed that nerve latencies responded similarly to subcutaneous temperature changes during and after ultrasound treatment. CONCLUSIONS: Alterations in nerve latencies from ultrasound on healthy nerves appeared to be related to temperature changes induced by ultrasound's thermal effects, and not by non-thermal or mechanical effects. PMID- 10812541 TI - Electrodiagnostic criteria in CIDP: comparison with diabetic neuropathy. AB - This study examines current criteria for CIDP by comparing the electrodiagnostic data from 17 patients who have biopsy proven CIDP with 29 patients with diabetic neuropathy. The groups were comparable in age. Data were examined using four sets of published criteria for primary demyelination. Two sets (A & B) defined demyelination based on distal motor latencies, F-wave latencies, motor conduction velocities, and temporal dispersion or conduction block. The other two sets defined demyelination based on slowing of motor conduction velocity < 70% of the lower limit of normal (LLN) in either 2 nerves (set C), or in a single nerve (set D). Using set A, 3/17 patients with CIDP met the criteria for a demyelinating neuropathy, while this was true in 2/29 of the patients with diabetic neuropathy. For sets B and C, data consistent with a demyelinating neuropathy were identified in 2/17 and 1/29 patients, respectively. Using set D criteria for demyelination was met in 6/17 of the patients with CIDP and in 5/29 of the diabetic patients. None of the criteria sets showed a significant difference between CIDP and diabetic neuropathy (p = 0.3 to 0.5). Though electrodiagnosis is an important element in the diagnosis of CIDP, current electrodiagnostic criteria alone are insufficient for defining many cases of CIDP. PMID- 10812542 TI - Recycling process of acetylcholine in the rat muscle controlled in a short-term memory manner. AB - The recycling process of acetylcholine (ACh) was studied in reference to muscle potentials evoked by sciatic stimuli in the presence of neostigmine. Wistar rats around 20 days old were anesthetized with urethane. Muscle potentials were recorded with concentric electrodes from their exposed medial gastrocnemius muscles before and after the injection of neostigmine. Attention was paid to changes in muscle potentials evoked by a set of 3 stimuli, and comparison was made mainly between successive two sets. Three potentials before neostigmine treatment were similar in amplitude. When a set of 3 stimuli was delivered 8 min after it, the first potential was as large as that before it. The others were greatly depressed. The first potential evoked by a set of 3 stimuli 10 min after neostigmine treatment showed a similar amplitude to the control level. The other potentials were again reduced severely. However, none of them were depressed so much when 3 stimuli were delivered 11 min after the drug application. When two sets of 3 stimuli were given with an interval of one minute, it was repeatedly observed in all animals that muscle potentials evoked by the second set were influenced by the first set. The second potential evoked by the second set was not so much depressed as seen when the interval was two or more minutes. Thus, a striking contrast was observed between successive two sets of 3 stimuli with an interval of one minute. These results indicate that the recycling process of ACh was activated following impulse transmission in rat muscles, and suggest that there exists such a neural mechanism as can exert an influence on the process in a short-term memory manner. PMID- 10812543 TI - Use of magnetic resonance imaging for the investigation of orbital disease in small animals. AB - Twenty-five small animal patients presenting with signs of orbital disease were investigated using magnetic resonance imaging (MRI) in an attempt to assess the value of this imaging technique for diagnosis. All patients were also examined using ultrasonography, and skull radiography was performed in 20 of these animals. The final diagnoses included neoplasia, inflammatory disease and foreign body penetration. MRI produced detailed images of orbital tissues and provided more information about the extent of pathology than the other imaging techniques; a correct diagnosis based solely on the MRI scan was made in 22 cases. Radiography was found to be helpful only in cases in which neoplastic disease extended markedly beyond the confines of the orbit into the nasal chamber and paranasal sinuses. Radiographic changes other than soft tissue swelling were not evident in other orbital disease processes. Ultrasonography gave both false negative and false positive diagnoses for neoplastic masses, although it allowed the correct diagnosis of both cases of foreign bodies and one of the three cases of retrobulbar abscesses in this series. MRI is recommended for patients in which radiography and ultrasonography fall to produce a confident diagnosis or for which surgery is proposed. PMID- 10812544 TI - Ultrasonographic examination of the adrenal gland and evaluation of the hypophyseal-adrenal axis in 20 cats. AB - The adrenal glands of 20 healthy, non-sedated cats were examined ultrasonographically; visualisation and assessment was possible in all cases. In comparison with the surrounding tissue, the adrenal glands were hypoechoic and two distinct zones could be differentiated in six of the cats. The length and width of the adrenal glands varied from 0.45 to 1.37 cm and 0.29 to 0.53 cm, respectively, and both dimensions could be reliably reproduced. The adrenal glands did not differ between male and female cats, and, in comparison to dogs, those of cats are more easily visualised ultrasonographically. The basal cortisol value ranged from 2.0 to 79 micrograms/litre. Values 30 and 60 minutes after administration of ACTH (0.125 mg/cat intramuscularly) varied from 36 to 126 micrograms/litre. The basal value of aldosterone ranged from 4 to 618 pg/ml. Values 30 and 60 minutes after administration of ACTH varied from 100 to 832 pg/ml. In all cats, suppression of the cortisol value below the level of detection (< 2.0 micrograms/litre) occurred four and eight hours after the administration of dexamethasone (0.1 mg/kg intravenously). PMID- 10812545 TI - Homonymous hemianopia in a pug with necrotising meningoencephalitis. AB - A 24-month-old female pug, which had previously been treated for visual hemifield loss, was referred with generalised seizures and other neurological disorders. A diagnosis of necrotising meningoencephalitis was suggested from the clinical signs together with the results of computed tomography and cerebrospinal fluid examination. This was confirmed seven months later by histological examination of the brain following euthanasia. Typical histopathological lesions of the disease were found in various areas of both cerebral hemispheres, including the visual striated cortex of the right cerebrum. PMID- 10812546 TI - Brain and ocular metastases from a transmissible venereal tumour in a dog. AB - A five-year-old crossbred dog was referred with rapidly growing masses over its penis and right popliteal lymph node. The dog had severe blepharospasm, congestion of episcleral vessels and rubeosis iridis of the left eye. A presumptive diagnosis of transmissible venereal tumour (TVT) and iridocyclitis was made based on the results of fine needle aspiration. Chemotherapy with vincristine and prednisolone was initiated and after four months the dog made a complete recovery. However, the dog subsequently relapsed, showing miosis, blepharospasm and a well defined mass within the anterior chamber of the left eye. In addition, the dog exhibited generalised 'grand mal' type seizures. Computed tomographic (CT) examination of the brain revealed two distinct masses in the left frontal lobe. Because of the poor prognosis, the owners elected to have the dog euthanased. On histopathology, metastases of TVT in the left eye and left cerebral hemisphere were found, showing no specific staining for CD3, immunoglobulin (Ig) G, IgM and lambda light chains. It was therefore concluded that the tumour growth was progressive, and that there was an absence of local humoral immune response against TVT in this case. PMID- 10812547 TI - Sprain of the short radial collateral ligament in a racing greyhound. AB - Severe carpal lameness in a racing greyhound due to a sprain of the straight part of the short radial collateral ligament is described. The dog subsequently developed an enthesiopathy at the origin of the ligament. Treatment was by kennel rest and the dog returned to successful racing. PMID- 10812548 TI - Echocardiographic diagnosis of dirofilariasis in a ferret. AB - Heartworm disease is described in a 14-month-old neutered female ferret (Mustela putorius furo) which had a three-week history of sneezing, anorexia and dyspnoea. Echocardiography revealed the presence of heartworms as hyperechoic densities within the right atrial and ventricular cavities. At necropsy, four Dirofilaria immitis parasites (three females, one male) were found in the right heart, the cranial vena cava and the caudal vena cava. Histopathological findings were similar to those reported in canine heartworm disease. Echocardiography may be a useful method of diagnosis of heartworm disease in the ferret. PMID- 10812549 TI - Aplasia of the gallbladder in a dog. AB - A young, female Maltese dog was presented with intermittent vomiting of bile. Biochemical evidence of persistent mild hepatopathy had been present for 11 months. Exploratory celiotomy was performed. Absence of the gallbladder with malformation of the quadrate lobe of the liver was identified. There was histological evidence of bile duct proliferation and portal fibrosis. PMID- 10812550 TI - Survey of companion animal diseases PMID- 10812551 TI - Feline injection-site fibrosarcomas: results of a BSAVA survey. PMID- 10812552 TI - [Inhaled nitric oxide therapy in preterm infants]. AB - According to a number of recently published, randomized controlled trials, treatment of persistent pulmonary hypertension of the newborn (PPHN) by the use of inhaled nitric oxide (iNO) has emerged as an established procedure in neonatology. The situation in premature infants appears to be more complicated than in the term infant. Due to the fact that nitric oxide interferes with platelet aggregation, the risk of intraventricular hemorrhage or its aggravation during iNO therapy is being discussed in a controversial manner. Since most studies are aimed at endpoints like oxygenation parameters, the presently available studies report extremely variable incidences of intraventricular hemorrhages (IVH). Meanwhile two large studies could demonstrate that clinical application of iNO in preterm infants is not associated with an increased incidence of IVH. Further randomized controlled trials of iNO in preterm neonates are highly desirable in order to establish the future role of this therapy and its indications. PMID- 10812553 TI - [Circulatory failure in children with left-to-right shunt in the framework of congenital heart defects: pathophysiology and therapeutic results]. AB - In children with large left-to-right shunts secondary to congenital heart defects the imbalance between the pulmonary and systemic perfusion may lead to circulatory congestion with clinical signs similar to those of heart failure. The circulatory function in this state was evaluated by using the invasive measurements performed during cardiac catheterisations in n = 64 young patients with ventricular septal defect (n = 56) or complete atrioventricular septal defect (n = 8) aging 0.1-23.7 years (median 1.1 years). The mean shunt ratio was Qp/Qs = 2.4 (range 1-8). With increasing shunt ratio the pulmonary perfusion raised (r = 0.84), but the systemic output dropped significantly (r = -0.77) while the total cardiac output (Qp + Qs) increased slightly not exceeding 141/min/m2. In infants, the systemic hypoperfusion affects the hemoglobin content: Hb = 14.9-1.01 x Qs, r = 0.63, p < 0.01. This may be due to the diminished oxygen extraction reserve of 46%. With dropping systemic output, the vascular resistance increases and the mean aortic pressure (MAP) remains normal. The actual pressure values layed near to the curve of the normal aortic pressure calculated as MAP = Qs x Rs. This pressure-flow-resistance diagram was used to interpret the effects of vasodilators established by 7 studies: ACE-Inhibitors, Hydralazine, and Na-Nitroprusside reduce the vascular resistance effectively but induce hypotension, because the systemic output fails to increase. In the chronic circulatory congestion secondary to a large intracardiac left-to-right shunt the pulmonary perfusion increases with the shunt ratio but the systemic output decreases and the total cardiac output is limited to a maximum of 141/min/m2. In this state vasodilators cause systemic hypotension thus offering no acceptable therapeutic option. PMID- 10812554 TI - [Minibrain/DYRK1A gene: candidate gene for mental retardation in Down's syndrome?]. AB - DYRK1A is the first member of a novel subfamily of protein kinases with dual specificity. The human gene for DYRK1A is located in the "Down syndrome critical region" (21q22.2). Due to its relationship to the Drosophila gene minibrain (Mnb), whose mutation results in specific defects in neurogenesis, and based on functional experiments on transgenic mice, DYRK1A is discussed as a candidate gene for mental retardation in Down syndrome. The kinase is characterized by its ability to catalyze tyrosine-directed autophosphorylation as well as phosphorylation of serine/threonine residues in substrates. Its exact cellular function is yet unknown. DYRK1A is, however, known to be translocated into the nucleus and supposed to be involved in the control of cell growth and development. The pathogenetic impact of DYRK1A on Down syndrome needs further elucidation. PMID- 10812555 TI - [Progressive familial intrahepatic cholestasis (Byler disease): current genetics and therapy]. AB - Progressive familial intrahepatic cholestasis (PFIC) is a congenital liver disease. First symptoms can frequently be seen shortly after birth. Quality and expectation of life are substantially reduced due to severe pruritus and the complications of progressive liver cirrhosis. PFIC is diagnosed on the basis of characteristic clinical and laboratory parameters and genetic analysis after exclusion of other liver diseases leading to intrahepatic cholestasis. Medical therapy is only effective in a proportion of children with PFIC. Partial biliary diversion (PBD) is nowadays considered the therapy of choice in patients with therapy-refractive pruritus. If performed in time, damage to the liver can be delayed or arrested, thus orthotopic liver transplantation (OLT) can be postponed or even avoided in at least some patients with PFIC. Besides providing a current overview of PFIC, we report on three patients who were successfully treated surgically. One patient was subjected to a new technique of PBD (cholecysto appendicostomy), the other two had OLT. PMID- 10812556 TI - [Neonatal progeroid syndrome (Wiedemann-Rautenstrauch syndrome): case report and review of the literature]. AB - The neonatal progeroid syndrome represents a complex of symptoms with unknown cause and pathogenesis. At 37 weeks of uncomplicated pregnancy a boy of non consanguineous turkish parents was delivered by section. The weight of 1740 g and the length of 43 cm were striking, the circumference of the head was still within normal range. Further on the patient showed the diagnostic criteria of Wiedemann Rautenstrauch syndrome: Beside growth failure and nearly absence of subcutaneous fat, old-looking face with hydrocephalic appearance, prominent scalp veins and sparse scalp hair, large hands and feet with long fingers and toes were found. Neonatal teeth didn't exist. But neonatal teeth are not always present in this syndrome. Among 19 published cases they were also not described in 6 cases (31.6%). Cataract was not found. A skin biopsy taken at the age of four months revealed an extremely thin dermis with pronounced rarefication of elastic material by light microscopy. On the electron microscopical level, no qualitative morphological aberrations or signs of degeneration were found, but normal although very small elastic fibres. With increasing age the appearance was nearly unchanged. In spite of high-caloric nutrition the increase of weight was not sufficient. The clinical symptoms of this patients were compared with the 19 already published cases of Wiedemann-Rautenstrauch syndrome. This shall be a contribution to the exact description of that extremely rare syndrome. We hope to facilitate, that hereby the differential diagnosis in difficult cases will be more easy. Because of heterogeneity it has to discuss if the Wiedemann Rautenstrauch syndrome really represents a separate genetic entity within the group of premature aging syndromes. PMID- 10812557 TI - [Arthrogryposis, renal tubular dysfunction, cholestasis (ARC) syndrome: case report and review of the literature]. AB - The ARC-syndrome is a rare disease with the obligatory symptoms arthrogryposis, renal tubular dysfunction and cholestasis. Optional further symptoms like ichthyosis, diarrhea, central nervous system defects and recurrent infections have been reported. The ARC-syndrome was first reported by Lutz-Richner and Landolt in 1973. The pathophysiology is still unknown, an autosomal recessive inheritance is postulated. Patients rarely exceed an age of six month. We report a boy of consanguineous Turkish parents who suffered from congenital deformities of the lower extremities, a metabolic acidosis and failure to thrive. In the sequel he developed a renal Fanconi syndrome and cholestasis. Histology of liver and muscle biopsy specimen showed the typical findings of the disease with giant cell hepatitis and neurogenous muscle atrophy. His condition could be stabilized and he increased in weight by substituting fluid, electrolytes, buffer and parenteral nutrition. Total enteral nutrition of the 280 ml/kg/d he required failed even by nasogastric tube and percutaneous endoscopic gastrostomy. Additional fluid substitution by central venous catheter remained necessary. At the age of 7 month he died. PMID- 10812558 TI - Pyogenic spondylitis with Scheuermann's disease. AB - We present the case of a teenager suffering from dorsal and lumbar pain. Imaging modalities diagnosed a lumbar retro-marginal anterior herniation and thoracic spondylitis. Such association has been reported twice in the literature. PMID- 10812559 TI - [Mollusca contagiosa in HIV-infected children receiving optimal antiretroviral therapy]. AB - Two children with symptomatic HIV-infection suffered from extended mollusca contagiosa. Intensified antiretroviral therapy including a protease inhibitor, resulted in a decrease of HIV RNA plasma concentration and a dramatic increase of CD4 T cells. Mollusca contagiosa nearly completely disappeared. These cases demonstrated that newly generated CD4 T cells during sufficient antiretroviral treatment have functional abilities. Therefore, sufficient antiretroviral treatment should be offered to HIV infected children with extended mollusca contagiosa before surgical intervention is considered. PMID- 10812560 TI - [Comment by the German Society of Pediatric Infectiology on a bulletin distributed by a laboratory service in Baden-Wurttemburg on Borrelia identification in ticks]. PMID- 10812561 TI - Clinician experiences of managed mental health care: a rereading of the threat. AB - The threat mental health professionals perceive in managed care, as indicated by their writings on the subject, is re-examined in light of evidence from an ethnographic study. Fieldwork focusing on clinician experiences of managed care was carried out at an urban community mental health center. Existing explanations of "the threat"--the possibility of deprofessionalization and the potential for deterioration in the quality of care--proved inadequate to account for the power it wielded at this site, perhaps because its full impact had yet to be felt at the time of data collection. A "rereading" suggests the meaning of managed care for this group of clinicians lies in the prospect of being gradually, unknowingly, and unwillingly reprofessionalized from critics into proponents simply by virtue of continuing to practice in a managed care context, and in losing a moral vision of good mental health treatment in the process. PMID- 10812562 TI - "Actually, I don't feel that bad": managing diabetes and the clinical encounter. AB - A major issue for persons treating and managing adult-onset diabetes (NIDDM) is the "problem of compliance." I consider the clinical encounter in the overall context of diabetes management as a punctuated experience focused on the cultivation of an ideal self whose "technologies" and "ethics of self-care" mimic a capitalist logic that links self-discipline, productivity, and health. Both clinicians and their patients share and identify with many of the cultural referents and social values that circulate through medical advice and practice. However, using individual examples, I show how this shared logic can produce idiosyncratic regimes of self-care and clinical practice that result in hybrid medical practices incorporating differing objectives and emphases concerned with a tolerable present or an ideal future. Rather than organizing principles for research and medical practice, I suggest that medical compliance and noncompliance should be considered part of the rhetoric to be explained within the regimes of a pursuit of health. PMID- 10812563 TI - Cultural variations in the placebo effect: ulcers, anxiety, and blood pressure. AB - An analysis of the control groups in double-blind trials of medicines demonstrates broad variation--from 0 to 100 percent--in placebo effectiveness rates for the same treatment for the same condition. In two cases considered here, drug healing rates covary with placebo healing rates; placebo healing is the ultimate and inescapable "complementary medicine." Several factors can account for the dramatic variation in placebo healing rates, including cultural ones. But because variation differs by illness, large placebo effects for one condition do not necessarily anticipate large placebo effects for other conditions as well. Deeper understanding of the intimate relationship between cultural and biological processes will require close ethnographic scrutiny of the meaningfulness of medical treatment in different societies. PMID- 10812564 TI - Menopause and the transmission of women's knowledge: African American and white women's perspectives. AB - Recent menopause literature does not sufficiently explore women's attitudes on the transmission of knowledge about menopause from sources other than biomedical providers. Analysis of 70 interviews with African American and Euro-American women shows that their perceptions of the intergenerational transfer of knowledge about menopause from their mothers shaped their attitudes toward menopause and the health-care technologies surrounding it. African American women who grew up in the segregated South frequently expressed that their mothers provided them with the knowledge and power to negotiate difficulties during the menopausal process, while many middle-class Euro-American women expressed that their mothers did not. Drawing on literature that examines the effects of race, class, and kinship on mother/daughter relationships, this article explores the reasons for this divergence. PMID- 10812565 TI - Concepts of illness and treatment practice in a caboclo community of the Lower Amazon. AB - Comparatively little has been written recently about the health consequences of social change and economic development in Amazonia. This study focuses on patterns of morbidity, treatment practices, and illness beliefs among caboclos of the Lower Amazon. It suggests that for these people traditional medicine is a salient marker of ethnic identity. An understanding of beliefs concerning disease etiology is critical to an appreciation of individual treatment choices in a plural medical system such as that found within the Lower Amazon region, where traditional healers can play a pivotal role in developing effective linkages to clinical services. PMID- 10812566 TI - [The "class effect" of drugs. Introduction]. PMID- 10812567 TI - [The "class effect" of the drugs]. PMID- 10812568 TI - [Angiotensin II antagonist receptors]. PMID- 10812569 TI - [Hypertensive cardiac remodeling]. PMID- 10812570 TI - [Vascular diseases and thrombosis]. PMID- 10812571 TI - [Clinical efficacy of losartan: are there any differences between angiotensin II type I receptor antagonists and other drugs?]. PMID- 10812572 TI - [Renal protection with antagonists of the angiotensin II receptors]. PMID- 10812573 TI - [Uric acid and arterial hypertension]. PMID- 10812574 TI - [Left ventricular hypertrophy regression in arterial hypertension. The efficacy of the angiotensin II type 1 receptor antagonists]. PMID- 10812575 TI - [Angiotensin II type 1 receptor antagonists in cardiac insufficiency]. PMID- 10812576 TI - The monosynaptic reflex: a tool to investigate motor control in humans. Interest and limits. AB - The principle of the monosynaptic reflex used as a tool to explore the excitability of the motoneurones (MNs) is explained and the general methodology of the H reflex is described. The different drawbacks inherent in the technique are then considered: mechanisms other than the monosynaptic la excitation of MNs contributing to the H reflex size (limitation of the H reflex size by disynaptic IPSPs, presynaptic inhibition of la terminals, post-activation depression); non linearity and changes in the 'recruitment gain' in the MN pool; and poor time resolution of the method. Despite these drawbacks, it is emphasized that the H reflex is the only available technique enabling one to investigate changes in transmission in spinal pathways during motor tasks. PMID- 10812577 TI - [Neurophysiologic study of tremor]. AB - Tremor is frequently encountered in neurologic practice. Clinical examination supplies information regarding its approximative frequency, regularity, amplitude, topography and activation conditions. The neurophysiological study helps in confirming the tremor, in differentiating it from other movement disorders like myoclonus, and may provide distinctive features which are important for the aetiological diagnosis. The neurophysiological investigation includes accelerometry, which analyses the mechanics of the movement. Spectral analysis with FFT allows the determination of frequency and amplitude. Accelerometry is always associated with surface EMG of at least two antagonistic muscles. It may show rhythmic bursts, their frequency, duration and activation pattern (alternating or synchronous). This neurophysiological approach to tremor has multiple interests. It may help in differentiating a true rest tremor from a postural tremor seemingly persisting at rest. Brief interruptions or rhythm breaks during distraction manoeuvers are seen in psychogenic tremors. Surface EMG may also demonstrate positive myoclonic bursts, or brief silent periods corresponding to negative myoclonus, sometimes pseudorhythmic, thus appearing clinically like a tremor, but investigations, aetiologies and treatment are different. Several features, especially slow frequency, may suggest a midbrain tremor, thus requiring brain imagery centered around the posterior fossa. Finally, the neurophysiological examination is the only way to demonstrate a primary writing tremor, or a primary orthostatic tremor, the frequency of which is pathognomonic while clinical symptoms are rather misleading. PMID- 10812578 TI - [Hypoxic-ischemic encephalopathy in the full-term newborn. Contribution of electroencephalography and MRI or computed tomography to its prognostic evaluation. Apropos of 26 cases]. AB - OBJECTIVES: Perinatal asphyxia complicated by hypoxic ischemic brain injury still remains the source of neurological lesions often serious and definitive. A major aim of neonatologists is to appreciate the severity of the hypoxic ischemic brain injury in the first days of life and to evaluate the forecast. The purpose of this work is to establish a relation between clinical signs, EEG, neuroimaging (MRI and CTS) and neuro-development. MATERIALS AND METHODS: 26 neonates from paediatric resuscitation unit (Hospital North, Marseille) were enrolled in a retrospective study since February 1994 to December 1997. All the newborns had at least one anamnestic criteria of perinatal asphyxia, an early electroencephalogram in the first two days of life and another between the third and the seventh day of life, and neuroimaging in the first 15 days of life: CTS in five cases and MRI in 21 cases. RESULTS: There was a good electro-clinic (P: 0.01) and prognostic (P: 0.03) correlation in patients within stage 3 of the "Sarnat classification". In the stage 2, the EEG did not provide valuable information about severity of the injury, and neuroimaging (especially MRI) allowed better prognosis. Diffuse brain injury or lesions of basal ganglia in MRI were associated, independently of clinical settings and EEG tracings, with a poor neurodevelopmental outcome (P: 0.02). The MRI was diagnostic in four cases of congenital encephalopathy complicated with neonatal suffering. CONCLUSION: Our study confirms the interest of the association of clinical settings, EEG tracings and MRI in the diagnosis and the prognostic of the hypoxic-ischemic encephalopathy in term neonate. PMID- 10812579 TI - Nonconvulsive status epilepticus: the role of morphine and its antagonist. AB - A 77-year-old woman with no history of epilepsy presented a probable nonconvulsive status epilepticus while receiving continuous intravenous morphine for back pain relating to vertebral metastasis of a malignant lymphoma. A generalized tonic-clonic seizure occurred a few minutes after injection of the morphine antagonist naloxone. No cerebral lesion was detected. This observation supports the notion of a possible pro-epileptogenic effect of opioid substances in association with metabolic and other toxic factors. Various hypotheses as to the complex actions of morphine and its antagonist in epileptogenesis are discussed. PMID- 10812580 TI - [Obstructive-sleep apnea syndrome: brain oxygenation measured with near-infrared spectroscopy. Preliminary results]. AB - This study assessed cerebral oxygenation in four obstructive sleep apnea syndrome (SAOS) patients (age = 51.8 +/- 15 years, apnea-hypopnea index = 68-125 per hour), during sleep and waking time, using near infrared spectoscopy (NIRS), during a standard polysomnography. Oxyhemoglobin (HbO2, reflecting cerebral oxygenation), total hemoglobin (Hbt, reflecting cerebral blood volumes) and cerebral oxygen saturation (SaO2c = HbO2/Hbt), were compared to the data obtained in four snorers not presenting apneas (age = 51.8 +/- 6.6, apnea-hypopnea index = 2.6-6.2 per hour) examined in the same way. The main result was that HbO2 values were reduced in SAOS patients, both during sleep (at stage 2: 52.54 +/- 9.60 mumol/L versus 73.80 +/- 11.70 mumol/L) and during waking state (53.67 +/- 7.20 mumol/L versus 63.05 +/- 5.55 mumol/L). Hbt was also reduced in apneic patients as compared to snorers during waking state (72.73 +/- 13.90 mumol/L versus 96.05 +/- 6.30 mumol/L). During sleep, Hbt increased in a similar way for snorers and apneics (12.4% versus 13%), whereas HbO2 values were constant for apneics. SaO2c paralleled SaO2p in snorers and apneics, values for SaO2c being 20-30% lower than values for SaO2p. The difference in the values was probably due to the use of different monitoring techniques. Cerebral oxygenation and cerebral blood volumes were continuously low in apneic patients, and peripheral hypoxia was associated with same-range cerebral hypoxia. Cerebral hemodynamic mechanisms related to sleep, although in part efficient in apneic patients, were not able to increase cerebral oxygenation up to normal values. PMID- 10812582 TI - Effect of apolipoprotein E genotype on lipid levels and response to diet in familial hypercholesterolemia. AB - The response of plasma lipids to dietary fat and cholesterol is partly genetically controlled. Apolipoprotein (Apo) E polymorphism has been shown to influence basal plasma lipid levels and the response to dietary changes in normolipidemic individuals. In general, subjects carrying the E4 allele have higher basal total and low density lipoprotein cholesterol (LDL-C) plasma levels and show an increased LDL-C response to dietary manipulation. The response to diet in subjects with familial hypercholesterolemia (FH) is also variable, but the influence of apo E genotypes on their dietary response has received little attention. We studied such influence on the lipid response to the National Cholesterol Education Program type I (NCEP-I) diet in 69 FH heterozygotes (44 women and 25 men). Subjects were studied at baseline (after consuming for 1 month a diet with 35% fat [10% saturated] and 300 mg cholesterol) and after 3 months of consuming a low-fat diet. No sex-related differences were found, and results were combined for men and women. The frequency distribution of apo E alleles was similar to that described in the general Spanish population: 0.0724 for the E2 allele, 0.0724 for E4 and 0.8551 for E3. Baseline plasma lipid and lipoprotein values were not influenced by apo E genotype. The response to the NCEP-I diet was similar in all subjects and no apoE allele-related differences were identified. As in non-FH subjects, there was a nonsignificant trend towards greater LDL-C lowering in E4 (-19.3%) than in E3 (-18.2%), and E2 (-16.6%) carriers. This finding supports the hypothesis that the impact of genetic defects at the low density lipoprotein receptor (LDLR) locus in FH subjects prevails over any influence on the part of ApoE polymorphism. PMID- 10812581 TI - Ciprofibrate effects on carbohydrate and lipid metabolism in type 2 diabetes mellitus subjects. AB - BACKGROUND AND AIMS: Atherosclerosis is the most common cause of morbidity and mortality in type 2 diabetes mellitus. Hyperglycemia, dyslipoproteinemia, arterial hypertension and coagulation abnormalities are the most important cardiovascular risk factors. Hypertriglyceridemia and low high density lipoprotein-cholesterol (HDLc) seem to be related with insulin resistance. Fibric acid derivates (fibrates) are effective in the treatment of dyslipoproteinemia in diabetes. Our aim was to evaluate the efficacy and safety of ciprofibrate in improving dyslipoproteinemia and its effect on fibrinogen plasma concentrations, carbohydrate metabolism variations and insulin action. METHODS AND RESULTS: 13 subjects with type 2 diabetes mellitus were treated with diet and placebo for 4 weeks and then randomized to one of two treatments: ciprofibrate 100 mg or placebo for four weeks. After a four-week wash-out period they were crossed over as shown in Figure 1. Total cholesterol, triglycerides, low density lipoprotein cholesterol (LDLc), very low density lipoprotein-cholesterol (VLDLc), HDLc, Apolipoprotein B100, fibrinogen, insulin and Lp(a) were measured. Insulin erythrocytes union was made by the Gambhir method. There was a 15% decrease in total cholesterol (p < 0.05) and 47% decrease in triglycerides (p < 0.01); similar changes were observed in VLDL-cholesterol and VLDL-triglycerides; 17% increase in HDL-cholesterol (p < 0.05). No significant differences were observed in LDL-cholesterol, apolipoprotein B100 and lipoprotein (a). Fibrinogen decreased 10% (p < 0.05). A non-significant 10% decrease in insulin secretion (area under curve) after oral glucose was observed with ciprofibrate. These findings indicate a decrease in receptor affinity. A non-significant decrease in insulin receptor number/cells was also observed. CONCLUSIONS: Ciprofibrate has a potent hypolipidemic effect, especially a decrease in triglycerides, VLDL and fibrinogen, and an increase in HDL-cholesterol, but does not influence glycemic control nor insulin action. Decreased insulin secretion may be due to peripheral use of glucose due to the drug's antilipolytic action. PMID- 10812583 TI - Relationship between apolipoprotein E polymorphism, postprandial hyperlipemia and hemostatic variables in patients with combined hyperlipemia. AB - BACKGROUND AND AIM: Combined hyperlipemia is a common lipid abnormality associated with coronary heart disease (CHD). Aim of this study was to investigate the relationship between this abnormality and hemostatic risk factors related to thrombosis. METHODS AND RESULTS: Forty-one patients were examined in the fasting state and during postprandial hyperlipemia. They had high levels of factor VIIc in the fasting state. During postprandial hyperlipemia a highly significant (p < 0.001) increase of activated factor (VIIa) occurred. This activation was not correlated to the increase in free fatty acids, but to the degree of postprandial hypertriglyceridemia. The absolute postprandial triglyceridemia was highest in subjects with the apolipoprotein (apo) E2 allele. Tissue factor pathway inhibitor activity (TFPIa) was correlated to LDL cholesterol and apo B concentrations and was highest in subjects with genotypes containing the E4 allele. CONCLUSIONS: Subjects with combined hyperlipemia may have an increased thrombotic risk related to activation of factor VII during postprandial hyperlipemia. Subjects with genotypes including the apoE4 allele may have a high lipid risk profile associated with high levels of LDL cholesterol. PMID- 10812584 TI - Cholesterol levels in Italian school children: results of an opportunistic survey. AB - BACKGROUND AND AIMS: A high prevalence of elevated serum total cholesterol (TC) levels has been described among Italian children. Universal screenings for TC have been suggested, though present recommendations are in favour of more selective and opportunistic surveys. Aim of the study was to assess TC distribution among 13-year-old schoolchildren in Central Italy. Further aim was to evaluate the indications for universal, selective or opportunistic screenings for TC. METHODS AND RESULTS: Children were examined opportunistically within a permanent screening programme for Mediterranean anemia carried out in the Lazio Region. TC was measured by dry chemistry in 3734 boys and 3644 girls aged 13 years from 77 schools in 37 municipalities. Results were grouped for 8 geographically and demographically similar areas. Mean TC was 137.6 mg/dl in boys and 144.3 mg/dl in girls. Desirable levels (< 170 mg/dl) were observed in about 90% of the children and fewer than 2% displayed levels indicative of genetic hypercholesterolemia (> 200 mg/dl). However, in two areas mainly populated by descendants from Northern Italy mean TC was remarkably high (158.7 and 152.2 mg/dl in boys and 164.5 and 160.0 in girls) and the percentage with desirable levels dropped to 68.6% and 74.7%. CONCLUSIONS: Our results show average good TC levels among Italian schoolchildren in Central Italy. This is probably due to their traditional Mediterranean diet. It can thus be suggested that only selective and/or opportunistic screenings for TC are indicated. However, in some areas where fewer children have desirable levels and almost 30% require dietary education, large-scale screenings are highly recommended. PMID- 10812585 TI - Triglyceride-lowering effect of omega-3 LC-polyunsaturated fatty acids--a review. AB - There is increasing evidence that serum triglycerides are a significant and independent risk factor for CVD. The aim of this report is to review recent literature pertinent to the triglyceride-lowering effect of omega-3 long chain polyunsaturated fatty acids (LC-PUFA). Animal data are not considered because they are difficult to extrapolate to the human situation. A large body of evidence derived from epidemiological studies and clinical trials has consistently demonstrated that this effect is dose-dependent and can be achieved by diet. The smallest amount of omega-3 LC-PUFA needed to significantly lower serum triglycerides appears to be approximately 1 g/day as provided by a fish diet. Use of fish oil administering as little as 0.21 g EPA and 0.12 g DHA per day significantly lowered serum triglycerides in hyperlipidemics. In normolipidemics, a daily intake of 0.17 g EPA and 0.11 g DHA, given as a fish oil supplement, induced a non-significant reduction of 22%. These findings must be considered as preliminary and warrant further research. Intake of omega-3 LC-PUFA is frequently reported to modestly increase LDL cholesterol. However, in normo- or slightly hyperlipidemic individuals who received omega-3 LC-PUFA for 4 months or longer, changes of LDL cholesterol were not significantly different from a placebo group. Both EPA and DHA lower serum triglycerides, but they may have a differential effect on lipoproteins. Intake of omega-3 LC-PUFA in the amount mentioned above is safe. PMID- 10812586 TI - Dietary antioxidants for cardiovascular prevention. AB - The generation of reactive oxygen species (ROS) is associated with life in aerobic conditions. ROS are thought to be implicated in the pathogenesis of various human diseases since they are capable of damaging biological macromolecules such as DNA, carbohydrates and proteins. The organism maintains defense against ROS, including enzymes and low molecular-weight antioxidants. An important source of antioxidants is diet which contains numerous compounds exhibiting antioxidant activity. A shortage of antioxidants in the diet might promote coronary heart disease through accumulation of oxidized LDL in macrophages. However, antioxidants may also influence endothelial functions, smooth muscle cell proliferation, thrombosis and plaque rupture. Consumption of fruits and vegetables, olive oil, red wine and tea is inversely correlated with heart disease rates. These foods are particularly rich in natural antioxidant nutrients, including ascorbate (vitamin C), the tocopherols (vitamin E) and carotenoids. More than 600 naturally occurring carotenoids have been identified. These compounds are plant pigments that provide the bright color of various fruits and vegetables; lycopene, which gives tomatoes their red color, is under active research. Flavonoids are > 4,000 naturally occurring substances which provide color, texture and taste for plant foods. As free radical scavengers, flavonoids inhibit lipid peroxidation, promote vascular relaxation and help prevent atherosclerosis. A sufficient supply with antioxidants from diet might help prevent or delay the occurrence of pathological changes associated with oxidative stress. When diet fails to meet the antioxidant requirement, dietary supplements might be indicated. The recently coined term nutriceuticals describes a variety of nonprescription products that are used to enhance health. The best known are vitamin E, vitamin C, carotenoids, coenzyme Q10, flavonoids and the amino acid L-arginine. Rigorous clinical trials, particularly among high-risk groups, are needed before they can be recommended routinely to patients. PMID- 10812587 TI - Osteoporosis and salt intake. AB - AIM: Recently, it has been hypothesized that salt intake may be related to the risk of osteoporosis. The aim of this review was to summarize the evidence for such relationship and to discuss possible mechanisms. DATA SYNTHESIS: We performed a review of the scientific literature on osteoporosis, particularly its etiology, and then focussed on studies addressing the relation between salt intake on the one hand, and calcium balance, bone resorption, bone mineral density and fractures on the other. Although a relation between high salt intake and increased bone loss is biologically plausible, the most pertinent studies relating salt intake to bone mineral density are only suggestive of high salt consumption as a risk factor for osteoporosis. Unfortunately, studies on fracture risk and salt intake are lacking. CONCLUSION: The relationship between salt intake and osteoporosis is still controversial. A possible relation between salt intake and fracture risk should be addressed in future studies. PMID- 10812588 TI - Chemical generation of excited states: the basis of chemiluminescence and bioluminescence. PMID- 10812590 TI - Physics of low light level detectors. PMID- 10812589 TI - Some brief notes on nomenclature and units and standards used in bioluminescence and chemiluminescence. PMID- 10812591 TI - Luminometer design and low light detection. PMID- 10812592 TI - Absolute calibration of luminometers with low-level light standards. PMID- 10812593 TI - Commercially available luminometers and low-level light imaging devices. PMID- 10812594 TI - Automated recordings of bioluminescence with special reference to the analysis of circadian rhythms. PMID- 10812595 TI - Chemiluminescence imaging systems for the analysis of macrosamples: microtiter format, blot membrane, and whole organs. PMID- 10812597 TI - Purification of luciferase subunits from recombinant sources. PMID- 10812596 TI - Overexpression of bacterial luciferase and purification from recombinant sources. PMID- 10812598 TI - A rapid chromatographic method to separate the subunits of bacterial luciferase in urea-containing buffer. PMID- 10812599 TI - Purification and ligand exchange protocols for antenna proteins from bioluminescent bacteria. PMID- 10812600 TI - Purification of firefly luciferase from recombinant sources. PMID- 10812601 TI - Chemical synthesis of firefly luciferin analogs and inhibitors. PMID- 10812602 TI - Structural basis for understanding spectral variations in green fluorescent protein. PMID- 10812603 TI - Large-scale purification of recombinant green fluorescent protein from Escherichia coli. PMID- 10812604 TI - Recombinant obelin: cloning and expression of cDNA purification, and characterization as a calcium indicator. PMID- 10812605 TI - Gonyaulax luciferase: gene structure, protein expression, and purification from recombinant sources. PMID- 10812606 TI - Dinoflagellate luciferin-binding protein. PMID- 10812607 TI - Assay of autoinducer activity with luminescent Escherichia coli sensor strains harboring a modified Vibrio fischeri lux regulon. PMID- 10812608 TI - Detection, purification, and structural elucidation of the acylhomoserine lactone inducer of Vibrio fischeri luminescence and other related molecules. PMID- 10812609 TI - Chemical synthesis of bacterial autoinducers and analogs. PMID- 10812610 TI - Overexpression of foreign proteins using the Vibrio fischeri lux control system. PMID- 10812611 TI - Application of bioluminescence and chemiluminescence in biomedical sciences. PMID- 10812612 TI - Use of firefly luciferase in ATP-related assays of biomass, enzymes, and metabolites. AB - The kinetics of ATP reagents not affected by product inhibition or other forms of inactivation of luciferase during the measurement time has been clarified. Under these conditions the decay rate of the light emission expressed as percentage per minute is a measure of luciferase activity and can be given as the rate constant k (min-1), directly reflecting the degradation of ATP in the luciferase reaction. Three types of reagents with different analytical characteristics and different application possibilities have been identified. Stable light-emitting reagents are suitable for measurements of ATP down to 1000 amol. This is the only type of reagent suitable for monitoring ATP-converting reactions, i.e., assays of enzymes or metabolites, assays of oxidative phosphorylation, photophosphorylation, and so on. A higher luciferase activity resulting in a slow decay of the light emission by approximately 10% per minute (k = 0.1 min-1) gives a reagent suitable for measurements down to 10-100 amol. The slow decay of light emission allows use of manual luminometers without reagent dispensers. A further increase of the luciferase activity resulting in a decay rate of approximately 235% per min (k = 2.35 min-1) and only 10% of the light emission remaining after 1 min is suitable for measurements down to 1 amol corresponding to half a bacterial cell. With this type of flash reagent the total light emission can be calculated from two measurements of the light intensity on the decay part of the light emission curve. This new measure is not affected by moderate variations in luciferase activity, but only by changes in quantum yield and self-absorption of the light in the sample. Flash-type reagents require the use of reagent dispensers. The stringent requirements for ATP-free cuvettes, pipette tips, and contamination free laboratory techniques make it unlikely that flash reagents would be useful in nonlaboratory surroundings. A potential application for this type of reagent is sterility testing. In general, it is concluded that one should select the ideal ATP reagent carefully for each application. Obviously the reagents used in a particular application do not have to match the decay rates given earlier exactly. However, various applications of the ATP technology and the properties of manual and automatic luminometers fall quite nicely into categories corresponding to the properties of the three reagents described. The rapidly growing interest in ATP technology has already resulted in the development of a greater variety of luminometers, from hand-held instruments to high-throughput systems. The continuation of efforts in both reagent and instrument development will undoubtedly result in many new applications. PMID- 10812613 TI - Chemiluminescent methods for detecting and quantitating enzyme activity. PMID- 10812614 TI - Chemiluminescence assay of serum alkaline phosphatase and phosphoprotein phosphatases. PMID- 10812615 TI - Chemiluminescence screening assays for erythrocytes and leukocytes in urine. PMID- 10812616 TI - Immunoassay protocol for quantitation of protein kinase activities. AB - Quantitation of at least two orders of magnitude of kinase enzyme concentration is achieved with detection of less than 0.1 U/well of src kinase activity (Fig. 3). A comparison between a sequential protocol, in which biotinylated peptide substance is captured prior to incubation with the kinase enzyme, and a simultaneous protocol, in which peptide capture and the kinase reaction proceed concurrently, demonstrates that the simpler simultaneous protocol provides similar detection sensitivity. these have also been demonstrated with 0.1 microM peptide substrate in a protein kinase A assay.5 Quantitation of protein kinase activity with chemiluminescent detection has been demonstrated with several different protein kinases, including both tyrosine and serine/threonine kinases.5 An immunoassay format provides high sensitivity and can be performed under conditions that most closely mimic physiological substrate and ATP concentrations with chemiluminescent detection. This assay format is also automated easily for use in high-throughput screening. PMID- 10812617 TI - Chemiluminescent immunodetection protocols with 1,2-dioxetane substrates. 4. AB - Chemiluminescent 1,2-dioxetane enzyme substrates provide a highly sensitive and versatile detection method for immunoblots and other membrane-based detections. 1,2-Dioxetane substrates, coupled with either alkaline phosphatase or beta galactosidase enzyme labels, generate glow light emission kinetics, with a signal duration that is significantly longer than most enhanced luminol/horseradish peroxidase chemiluminescent detection systems. The long-lived, high-intensity light signal is ideal for imaging using a variety of formats, including X-ray film, photographic film, chemiluminescence phosphor imaging screens, and the rapidly expanding selection of camera imaging systems. PMID- 10812618 TI - Chemiluminescent reporter gene assays with 1,2-dioxetane enzyme substrates. AB - 1,2-Dioxetane chemiluminescent substrates provide highly sensitive, quantitative detection with simple, rapid assay formats for the detection of reporter enzymes that are widely used in gene expression studies. Chemiluminescent detection methodologies typically provide up to 100-1000x higher sensitivities than can be achieved with the corresponding fluorescent or colorimetric enzyme substrates. The varieties of 1,2-dioxetane substrates available provides assay versatility, allowing optimization of assay formats with the available instrumentation, and are ideal for use in gene expression assays performed in both biomedical and pharmaceutical research. These assays are amenable to automation with a broad range of instrumentation for high throughput compound screening. PMID- 10812619 TI - Clinical application of Southern blot hybridization with chemiluminescence detection. PMID- 10812620 TI - Quantitative polymerase chain reaction and solid-phase capture nucleic acid detection. AB - The combination of PCR amplification and chemiluminescent detection of PCR products provides a highly sensitive system for the quantitation of DNA and RNA. The broad dynamic range of the chemiluminescent detection assay simplifies the selection of cycling and concentration parameters critical to harnessing the quantitative aspects of PCR amplification. Detection of 200 amol of PCR product is attained using the described procedures. The tube or microplate format of the assay avoids many of the limitations associated with other methods of PCR quantitation involving gel electrophoresis. This detection methodology can be applied to a variety of quantitative nucleic acid assays, including viral load and gene expression analysis. PMID- 10812621 TI - Targeted bioluminescent indicators in living cells. PMID- 10812622 TI - Green fluorescent protein as a reporter of transcriptional activity in a prokaryotic system. PMID- 10812623 TI - Bacterial lux genes as reporters in cyanobacteria. PMID- 10812624 TI - Application of bioluminescence to the study of circadian rhythms in cyanobacteria. PMID- 10812625 TI - Construction of lux bacteriophages and the determination of specific bacteria and their antibiotic sensitivities. PMID- 10812626 TI - Luciferase gene as reporter: comparison with the CAT gene and use in transfection and microinjection of mammalian cells. PMID- 10812627 TI - In situ hybridization and immunohistochemistry with enzyme-triggered chemiluminescent probes. PMID- 10812628 TI - Blood phagocyte luminescence: gauging systemic immune activation. PMID- 10812629 TI - Demonstrations of chemiluminescence. PMID- 10812630 TI - Bioluminescence as a classroom tool for scientist volunteers. AB - There is a great need for practicing scientists to volunteer their time and expertise in the K-12th grade science classroom. We have found that bioluminescence is a fun and exciting way to teach basic science concepts and is an excellent tool for the volunteering scientist. We have had very positive reactions from both teachers and students. The excitement of the students when they first see bioluminescence is contagious. Bioluminescent dinoflagellates are one of the easiest ways to introduce students to this fascinating topic. Many activities and experiments can be done using the bioluminescent dinoflagellates and many students and teachers could benefit from your knowledge and expertise. See you in the classroom. PMID- 10812631 TI - Green fluorescent protein in biotechnology education. PMID- 10812632 TI - [Why history of psychiatry?]. PMID- 10812633 TI - [The continuity of biological models in psychiatry: melancholy as brain disease in scholastic psychology]. AB - This paper emphasizes the long lines of tradition biological concepts have in the explanation of mental disorders in classical antiquity and the middle ages; yet it also portrays rivalling theories. I describe both medical and philosophical approaches, as they merged into a quite coherent model during scholasticism. The work of Albert the Great serves as an example for this model. A last paragraph shows its plausibility for authors even of the 17th century and discusses the epistomological basis of dualistic systems. PMID- 10812634 TI - [Karl Ludwig Kahlbaum (1828-1899)--a protagonist in modern psychiatry]. AB - Kahlbaum was one of the most influencial psychiatrists of the 19th century. He paved the way for Kraepelin and inspired the development of 20th century clinical psychiatry. With his work he contributed to new ideas in general psychopathology and in psychiatric nosology. He described "clinical state-course-entities" and developed the concept of "exogenous reaction types", which would later be known as the "Bonhoeffer-paradigm". Kahlbaum was also the first to differentiate between centripetal, intracentral and centrifugal psychic functions, which was later transformed into the "psychic reflex circuit" by Wernicke. The concept of organic versus non-organic psychoses proved to be a heuristically fruitful classification system even for modern psychiatry. Kahlbaum was the first German psychiatrist to describe several disorders and syndromes and to coin new psychiatric terminology for them, including paraphrenia, hebephrenia, catatonia and cyclothymia. These terms and their originally associated meaning are still in use today. Kahlbaum's ambitious scientific aim was to develop specific therapeutic strategies which were based on a methodologically and etiologically sound classification system. PMID- 10812635 TI - [Emil Kraepelin's reviews for the "Literarische Centralblatt fur Deutschland"]. AB - For about 10 years, namely from 1882 until 1892/93 Emil Kraepelin worked as a reviewer for the "Literarische Centralblatt fur Deutschland", probably the most influential review journal of that time in Germany. In these short essays their author had to state his opinion on many different topics such as anthropology, (experimental) psychology, various aspects of the history of psychiatry as well as its different schools like the so-called "psychiatry of brain" in the most succint way. Also they reveal personal feelings and opinions Kraepelin had about some of his contemporaries. Last but not least they can also demonstrate in which way his attitude towards a particular subject changed over the years. Additionally employing original letters written to the editor of the "Literarische Centralblatt" as well as other primary sources a quite clear picture of the young Kraepelin as man and scientist can be drawn. That is the objective of this paper as well as bringing those nowadays almost forgotten works, which cannot be found in any of the profound bibliographies on Kraepelin, back to the attention of the psychiatrical public. PMID- 10812636 TI - ["Not unaffected by northern winds"? Swiss psychiatry and eugenics in the period between world warms]. AB - On the basis of the publications and meeting reports of the Swiss Society of Psychiatry this essay examines the relationship of Swiss psychiatry and eugenics during the time between the wars. In a democratic country Swiss psychiatry, on the one hand held a leading position in the justification of eugenic measures, on the other hand a majority of Swiss psychiatrists refused coercive measures, and in World War II disassociated from the racialist eugenics and healthpolicy that were established in NS-Germany. Thus, in difference to NS-Germany the practice of eugenic sterilizations in Switzerland wasn't regulated by any specific national law. PMID- 10812637 TI - [Murder of psychiatric patients and destruction of psychiatric culture. The Leipzig-Dosen Saxony state asylum in the first half of the 20th century]. AB - OBJECTIVE: The paper examines the development of the Saxonian state mental asylum Leipzig-Dosen in the first half of the twentieth century. METHODS: A rich store of administrative files and case registers allowed us to demonstrate the asylum's history in the field of tension between therapeutic optimism and medical failure. RESULTS: Dosen integrated important structural and therapeutic reforms, had to take care of a rising number of patients with little effective treatment measures and decreasing economic resources and it was involved in most of the inhuman deformities of psychiatry during the period of National Socialism. CONCLUSIONS: The asylums development can be considered as an example for the breakdown of the psychiatric culture in Germany. PMID- 10812638 TI - [Open care--the Rodewisch theses--psychiatry enquete: 3 reform approaches compared]. AB - AIM: The three most important German approaches for reforming psychiatric care in the 20th century--i.e. "open care" in the Weimer Republic in the 20ies, the "Rodewisch Theses" published in the GDR 1963, and the "Psychiatry-Enquete" in the Federal Republic in 1975--were compared using seven criteria. METHODS: Psychiatric and general medical journals were systematically searched for articles on aims and objectives of mental health care. RESULTS: All three approaches promoted stronger community based care. None of them suggested an "empowerment" as it is understood today. Differences between them concerned equal care for mentally and physically ill patients, international relations, and economic arguments. Comparisons on other criteria, e.g. the relationship with university psychiatry, yield a less clear picture. CONCLUSIONS: All three ideas for reforms have been successful--to a limited and different extent. They resemble each other in essential principles, but there are also substantial differences. PMID- 10812639 TI - [Everything has a beginning. Comments on the historical development of the psychiatry enquete]. PMID- 10812640 TI - [Friedrich Nietzsche: history of his illness. On the 100th anniversary of the death of the poet-philosopher]. AB - The philosopher Friedrich Nietzsche was one of the most famous patients in the psychiatric hospital Jena. Between 1889 and 1890, Otto Binswanger und Theodor Ziehen treated the progressive paralysis of Friedrich Nietzsche whose 100th anniversary of death is on 25th August 2000. After Nietzsche's death, an animated discussion about his disease arose. Many objected against the syphilitic diagnosis of the disease. Nietzsche's medical fill became a popular object of science, although it ended as a very odd story. PMID- 10812641 TI - [Neurological complications of HIV infection. Review: new diagnostic, therapeutic and prognostic aspects]. AB - About one third of patients with HIV infection show neurological complications with considerable morbidity and high mortality. This is an actualized review of the most important neurological manifestations resulting from primary HIV infection, from secondary opportunistic infections, or as complications of antiretroviral therapy. The primary neurological manifestations, including HIV associated dementia complex, myelopathies, peripheral neuropathies and myopathies, the more common opportunistic infections, primary central nervous system lymphoma and cerebrovascular diseases, are discussed in the light of new evidence in diagnosis, therapy and prognosis. Cognitive and psychiatric symptoms, visual changes, headache, seizures, dizziness, involuntary movements, gait disturbances, cranial neuropathies and focal deficits are the common neurological symptoms in HIV infection which are described under the aspect of differential diagnosis. It is important to bear in mind that nearly all information available to date on this subject concerns HIV patients in the period before combination therapies (including protease inhibitors). The introduction of highly active antiretroviral therapy (HAART) with protease inhibitors in 1995, and non nucleoside reverse transcriptase inhibitors, have opened up new therapeutic modalities with a new emphasis on earlier detection and treatment of neurological complications. The prognosis of different HIV-associated neurological diseases has considerably improved, as recently shown in the case, for example, of progressive multifocal leucoencephalopathy. PMID- 10812642 TI - [Hemoglobin and fatigue in cancer patients: inseparable twins?]. AB - Tiredness is one of the most common symptoms in patients with cancer, and, because of its multifaceted manifestation in this population today, it is generally termed the "fatigue syndrome". Causal relationships are complex and multidimensional. Research suggests that the type and stage of cancer play an important role, and that side effects of cancer treatment as well as psychological factors may be involved. A causal relationship between fatigue and a certain degree of anaemia has been suggested. Whilst it is acknowledged that severe anaemia causes "fatigue", the influence of mild, chronic anaemia on quality of life is not yet clear. It is the aim of this prospective, descriptive study to investigate the correlation between "fatigue" in cancer patients and their haemoglobin levels. Haemoglobin was measured in 444 cancer patients with different types and stages of cancer and treatment modalities. Concurrently, fatigue was assessed with the new Fatigue Assessment Questionnaire (FAQ). In addition, the Karnofsky Index was assessed and depression and anxiety were measured with the Hospital Anxiety and Depression Scale. RESULTS: Cancer patients with a haemoglobin level below 11 g/dl suffered from fatigue more than patients with higher haemoglobin levels (r = -0.196, p < 0.001). The correlation of "fatigue" was most prominent with physical fatigue such as reduced physical performance or weakness. The Karnofsky Index correlated significantly with the haemoglobin concentration. Patients with lower haemoglobin levels showed more signs of depression than those with higher levels. CONCLUSIONS: The grade of anaemia correlates with intensity of fatigue in cancer patients and also with depression. Both can be related to the advanced disease situation. Data suggest that if levels of haemoglobin are lower than 11 g/dl, subjective fatigue should be specifically assessed to plan treatment according to the patients' need. It remains to be substantiated when and which treatment is indicated to alleviate fatigue and to support quality of life in the individual anaemic cancer patient. PMID- 10812643 TI - [Autologous transplantation of peripheral blood hematopoietic stem cells in the treatment of hematological malignancies. I: patients]. AB - 61 autologous transplantations for haematological malignancies have been performed with peripheral blood stem cells in 60 patients. 26 grafts were performed for non-Hodgkin's lymphoma (18 patients were transplanted in first remission, 8 patients after progression or relapse), 13 for multiple myeloma, 7 for Hodgkin's disease and 10 for acute myeloid leukaemia. One patient died from thrombosis of the portal vein. 38 patients were in complete remission 29 months (extremes: 14-44) after transplantation. 21 of 60 patients progressed or relapsed after transplantation, and 14 died. No death was attributed to graft failure, infection or haemorrhage. In conclusion, transplantation with peripheral blood stem cells is well tolerated, has a low toxicity rate, and can be used safely for patients with haematological malignancies. PMID- 10812644 TI - [Hereditary hearing loss: new diagnostic possibilities]. AB - Mutations in many different genes can result in hearing loss. Using different molecular genetic methods, the disease-causing gene mutations can often be identified or at least localised to defined regions of the genome. These new diagnostic possibilities result from the localisation and identification of a number of hearing-loss genes in the last five years. Diagnostic investigations should always be accompanied by a genetic counselling of the family. In addition, the isolation thus far of 11 genes mutated in autosomal dominant inherited hearing loss, as well as of 6 genes mutated in autosomal recessive inherited hearing loss, has contributed to a better understanding of the molecular pathology of hearing loss in general. However, we are only beginning to see the whole picture, as an estimated 50 to 80 hearing loss genes remain to be discovered. PMID- 10812645 TI - [Unusual cause of pathologic echo-structure of the liver]. PMID- 10812646 TI - [Further evidence for the renoprotective effect of ACE inhibitors: ramipril protects against the progression of chronic renal insufficiency in non-diabetic nephropathy with nephrotic proteinuria]. PMID- 10812647 TI - [Triage of acute coronary syndromes: is an additional biochemical marker necessary?]. PMID- 10812648 TI - Case of the season. Frontal chest radiograph demonstrates marked hyperinflation and bilateral chronic infective changes with the typical appearance of long standing pulmonary cystic fibrosis. PMID- 10812649 TI - An overview of pulmonary host defense. AB - In response to a constant barrage of inhaled and aspirated material, the lung has evolved a sophisticated multi-tiered system of defense. Components of this system include mechanical and biochemical barriers, phagocytic scavengers, and specific cellular and humoral immune responses. A breakdown of individual components of this system is frequently associated with the inability to maintain a sterile milieu and the consequent development of respiratory tract infection. PMID- 10812650 TI - Pulmonary infections in HIV/AIDS. PMID- 10812651 TI - Pulmonary infections in cancer and bone marrow transplant patients. PMID- 10812652 TI - Pulmonary infections in patients who have received solid organ transplants. PMID- 10812653 TI - The spectrum of pulmonary infection in the immunocompromised child. PMID- 10812655 TI - [Laparoscopic ovariectomy in the standing mare]. AB - An eleven year old warmblood mare was referred to Dierenkliniek Emmeloord because of recurrent cyclic behavioral problems. A standing bilateral laparoscopic ovariectomy was performed. Due to this minimal invasive technique a quick return to training was possible. PMID- 10812654 TI - Mild immune deficiency and its consequences. PMID- 10812657 TI - [Ultimate pre-transfusion control: evaluation of seven appliances]. AB - The transfusion unit of the Institut Gustave Roussy has tested seven pre transfusion ABO control devices registered at the Agence francaise de securite sanitaire et des produits de sante. Determination of the optimal plan to replace the existing plan in our institution was the primary objective of this study. A significant heterogeneity was observed among tested devices. None of the tested plans fulfilled all the desired quality criteria. PMID- 10812656 TI - [Preservation of erythrocytes of heterozygous SC sickle cell patients]. AB - In order to evaluate the feasibility of the autologous transfusion in an alloimmunized sickle cell patient, changes in the hematologic and biochemical characteristics of erythrocytes stored for 42 days from two patients with sickle cell SC anemia were compared with control subjects' (Hb A) red blood cells. Erythrocytes were stored in Saline Adenosine Dextrose Mannitol at +4 degrees C. The cryopreservation storage was made and 51Cr red cell survival was measured in one patient. No significant difference in the hematologic and biochemical parameters of the SC red blood cells and the control subjects was observed during the storage at +4 degrees C. Red cell survivals determined in fresh cells, cells stored for 42 days at +4 degrees C and thawed cells from one patient demonstrate much shorter half-life values than those of normal red blood cells. Before application, our results need to be confirmed by the same protocol with another patient with sickle cell SC. PMID- 10812658 TI - [Computer-based hospital transfusion process: why and how does one establish a link between continuous quality improvement and clinical information system?]. AB - Within a hospital, the need for a computer-based transfusion system has became mandatory. It facilitates the tracing of healthcare activities, which is the basis of the security of the care and a functional element of continuous quality improvement procedures. In order to implement this traceability, reactive and real-time information systems are needed close to healthcare participants, which is not the case of current information systems which rely on a recorded collection of data, far from the needs of the caregiver, and mainly answering to an objective evaluation of results. In the context of continuous quality improvement programmes started in our hospital, hemovigilance was the first to use a process analysis approach, from the prescription of blood units to their administration and follow-up. Several questions arise from this: 1) how to use the process analysis work to specify the users' needs of a generalized and real time transfusion information system? 2) how to spread this model to other healthcare activities? 3) how to integrate or interface the whole of these quality programmes with a clinical information system? A user-centered methodology was used, based on 'usage cases'. For each step of the transfusion process, this method allowed us to specify participants, data necessary for an activity (observed, deduced or decision-support data), data issuing from the activity, roles (the interaction between user and activity) and functions (the result of the interaction between user and activity). PMID- 10812659 TI - [Screening for markers of infections transmitted by transfusions in the blood collected in France from 1996 to 1998]. AB - From 1996 to 1998, a decrease in positive donation rates has been observed for HIV, HCV and HBs Ag in first-time donors, while these rates remained stable for HTLV. In repeat donors, the same decrease was observed for HCV and HBs Ag while the rates remained stable for HIV. No HTLV-positive donations from repeat donors were noted in 1998. About half of the HIV-positive repeat donors were regular donors (less than two years between the two donations), as well as 88% of HBV infected repeat donors. Inversely, only 20% of HCV-positive repeat donors were regular donors. Anti-HBc antibodies have been found in 20% of HIV-infected donors, in 22% of HCV-infected donors, and were associated with HBs Ag in 99% of the cases. Elevated ALT was observed in 47% of donors with anti-HCV and in 10% of donors with HBs Ag. The major risk factors are at-risk sexual behavior for HIV and use of intravenous drugs and nosocomial infections for HCV. Being a native of an endemic country has been found to be the major risk for HBV. The major HTLV risk factor was directly or indirectly linked to the Caribbean area. PMID- 10812660 TI - [Prevalence of bacterial contamination of standard platelet units: prospective study]. AB - BACKGROUND: Platelet concentrates contaminated with bacteria are a main source of transfusion-associated sepsis. Several studies have reported a very wide incidence (0-10%) of contamination. The aim of this study is to assess the prevalence of bacterial contamination of standard platelet units at the regional blood transfusion center in Casablanca. STUDY DESIGN AND METHODS: During 15.5 months, 3,458 platelet concentrates (PCs) were analysed. The plasma from the bag and the tube was homogenised. One-fifth of the tube contents were inoculated every day for five days in two tubes containing 2.5 mL of Tryptic soy broth. One tube was incubated at room temperature and the other at 37 degrees C for 24 hours. The bacteria were identified by standard procedures. RESULTS: Twenty-five (0.72%) of 3,458 platelet bags were contaminated. Twenty-two bacteria (88%) were found in the platelet bags stored three days or more, of which 14 (56%) were Staphylococci coagulase negative, two Staphylococcus aureus, one alpha-haemolytic Streptococcus, four bacillus cereus, one Enterobacter cloacae, one Escherichia Coli and two Xanthomonas maltophilia. CONCLUSION: The incidence reported in this series is higher than that reported by most authors. PMID- 10812661 TI - [Platelet transfusion and allergic transfusion reactions: experiences at Lille Hospital over a four year period]. AB - Among the immediate transfusion reactions caused by the utilization of blood products, those suggesting immuno-allergic mechanisms posed problems for frequency, gravity, laboratory diagnosis and safety. We report here the Lille Hospital's experience over a four-year period concerning these manifestations after platelet concentrate transfusion. Eight hundred and fifty-two immediate transfusion reactions have been declared, of which 230 were allergic, which appeared in 181 patients (27%). Among the most frequent clinical signs, rash was often described (158 cases: 68.7%); less frequent were respiratory problems such as dyspnea (34 cases: 14.8%) and hypotensive reactions (18 cases: 7.8%). Seven patients presented severe reactions (3%). Twenty percent of them presented multiple allergic reactions and in 43.2%, the recurrence was more serious than the initial problem in spite of preventive medication; the use of washed blood components was necessary. The age of platelet concentrates does not appear to play a part in provoking these events (67% of platelet concentrates had been collected within four days). These allergic transfusion reactions posed problems for those who prescribe medication, because they are frequent, sometimes serious, can recur and at present, the proposed medication prevention is not always efficient. PMID- 10812662 TI - [Quality indicators and dashboards: the experience of a collection department]. AB - Production of labile blood products and pharmaceutical products derived from blood must meet quality and safety requirements set and controlled by regulatory provisions (Guidelines for Transfusion Practices, Characteristics of Labile Blood Products). With its activities exerted at the start of the transfusional chain, collection departments have to meet internal requirements (self-sufficiency, procurement regulation, compliance of blood products as raw materials) as well as external ones (donor and recipient safety). To fit in a dynamics of on-going quality improvement, the quality approach of a department must implement a system to monitor and control its activities. The subject of this work is to report an experience of the implementation of quality indicators and specific dash boards for whole blood collection and related activities in a blood transfusion center. PMID- 10812663 TI - [The combination of chronic poliomyelitis and syringomyelia. 1902]. PMID- 10812664 TI - [Neuropsychological syndromes of arteriovenous malformation of cinguli gyrus and hippocampus]. AB - 41 patients with arteriovenous malformations (AVM) of gyrus cinguli and 39 ones with AVM of hippocamp were examined. AVM verification was performed according to the data of angiography and computer tomography of the brain during the operation. 4 patients had generalized tonic-clonic seizures and absans. In these cases AVM spread from gyrus cinguli and hippocamp to the cortical medio-basal regions of frontal and temporal lobes of the brain. Autonomic paroxysms weren't found in the patients. Before the operation 73 patients had disorders of memory that progressed after the operation. Disorders of memory weren't completely identical in damages of gyrus cinguli or of hippocamp, namely: in damages of gyrus cinguli such disorders were similar in terms of their characteristics with those observed in damages of medio-basal regions of the frontal lobes, while in pathology of hippocamp--with those of diencephalic region (region of III ventricle). A conclusion was made about different participation of the structures studied in realization of the memory function. PMID- 10812665 TI - [Neurological and psychopathological syndromes in the follow-up period after exposure to ionizing radiation]. AB - Neurologic, psychiatric and psychophysiologic (computed EEG) examinations were carried out in 100 Chernobyl accident's survivors who had got acute radiation sickness (ARS), in 100 Chernobyl liquidators who worked for 5 or more years in the zone (1986-1987) as well as in control groups: 20 normal age- and gender matched adults and 50 veterans of the Afganistan war with consequences of the posttraumatic stress disorder (PTSD) and 50 veterans with both PTSD and mild closed head injury. Left-hemispheric cortical-limbic and diencephalic right hemispheric syndromes were revealed. Left-hemispheric frontal-temporal-limbic dysfunction was associated with schizophrenia-like syndrome, while diencephalic right-hemispheric dysfunction--with the affective syndrome. Doses more than 0.3 Sv (including the ARS-patients) resulted more frequently in the left-hemispheric cortical-limbic and schizophrenia-like syndromes. Diencephalic right-hemispheric and affective syndromes were more frequently observed after the exposure to doses less [corrected] than 0.3 Sv. Development of schizophrenia spectrum disorders in the irradiated Chernobyl survivors could be due to radiation-induced left fronto temporal-limbic dysfunction following irradiation doses more than 0.3 Sv (including the ARS-patients). The cerebral patterns of schizophrenia and postradiation brain damage are similar. Persons exposed fo 0.3 Sv and more could be classified as the group of higher risk of the development of schizophrenia spectrum disorders. The authors suggest that ionizing radiation may be an environmental trigger factor which can cause schizophrenia in the predisposed subjects. PMID- 10812666 TI - [The influence of hormones of stress-promoting system on the course of acute ischemic stroke]. AB - Experimental and clinical studies have proved an important role of the reaction of a stress-realizing endocrine system for both a course and an outcome of acute somatic diseases. That is why plasma concentrations of thyroxine (T4), triiodo thyronine (T3), thyrotropic (TTH) and adrenocorticotropic (ACTH) hormones as well as of renin were measured during the 1-st week of acute ischemic stroke (AIS) in relevant patients for determination of their prognostic significance. 16 patients with hemispheric AIS were examined. A complex clinical biochemical investigation performed in patients with acute disorders of cerebral circulation revealed the presence of the "low T3 syndrome" accompanied by an increase of blood plasma levels of T4 by the 7-th day of the disease as well as an increase of the concentrations of TTH, ACTH and renin on the 2-nd day of the stroke. That reflected the severity of cerebral ischemia. Close correlation observed between the degree of normalization of hormonal concentrations and manifestations of neurologic disorders' regression by the 7-th and the 21-st days gave a good ground to consider these indices as a prognostic criterion for determination of an outcome of the acute period of ischemic stroke. PMID- 10812667 TI - [The administration of dipyridamole (curantil) for the secondary prevention of ischemic stroke]. PMID- 10812668 TI - [Autonomic disorders in persons with asthenic syndrome and their correction with citrulline malate]. AB - The paper presents the results of the investigation of psychoautonomic correlations in 15 patients with psychogenic asthenia. Spilberg's and Beck's tests as well as autonomic questionnaire were used for the psychologic testing. Sympathic-parasympathic correlations were investigated by means of cardiovascular tests, by evoked skin sympathetic potentials, variability of cardiac rhythm under conditions of different functional states. The patients had mild anxious depressive and pronounced autonomic disorders with prevalence of cerebral sympathic-adrenal impacts, elevation of vagal influences in cardiovascular system, mediatory insufficiency of the sympathic sweating nerves and a decrease of the adaptive abilities of the regulatory systems. To correct the asthenic manifestations citrullin malate (stimol), a metabolic corrector produced by "Laboratories BIOCODEX" (France) was used. Stimol relieves psychoautonomic disorders by increasing both power capacities of the cells and synthesis of biologically active substances. PMID- 10812669 TI - [The experience of application of olanzapine: an atypical neuroleptic in acute schizophrenia]. AB - Thirty adult patients with acute schizophrenia were included into six-week open labelled, non-comparative trial of olanzapine with free dosing from 5 to 20 mg per day. For assessment of efficacy and safety of the treatment PANSS, BPRS, CGL and ESRS scales were used. The main criterion of improvement was the percentage of reduction of BPRS score to the end of the trial. More than 50% reduction was achieved in 23% of the patients, more than 20%--in 57% of the cases; 20% were non responders (reduction less than 20%). Changes in the scores of the positive and negative PANSS subscales were significant (p < 0.001) starting from the second week of treatment. There was no correlation in changes in positive and negative scores as well as in changes in negative and ESRS scores. The first signs of the antipsychotic effect (a reduction of tension, suspiciousness and fear) appeared just in the first two weeks of treatment and manifested in the reduction of delusions and hallucinations. Reduction of depression didn't correlate with changes in scores of the positive or negative PANSS subscales. The changes in behavioral and cognitive symptoms were statistically significant starting from the second week of treatment. No clinically significant signs of extrapiramidal syndrome or other side-effects except weight gain were observed during the trial. PMID- 10812670 TI - [Treatment of anxiety-related depression with tianeptine (coaxil) in patients with ischemic heart disease]. PMID- 10812671 TI - [Recovery process after the diphtheric polyneuropathy according to interference electromyography]. AB - In order to control the recovery process after diphtheric polyneuropathy the dynamics of activity of m.flexor carpi radialis with increasing force was repetitively investigated in 12 patients using surface electromyography (EMG). In patients examined 12.4 +/- 2.9 months after the onset of diphtheria neurogenic type of EMG (decreased amplitude and number of turns along with increasing force) was established. 11-14 months later clinical improvement followed by positive changes of EMG parameters was documented. PMID- 10812672 TI - [Pathogenetic mechanisms of stuttering]. AB - A complex study including neurologic, psychologic, neurophysiologic and EEG investigation of 33 stutter patients was performed to determine pathogenetic mechanisms of stutter. All clinical data obtained were presented by means of structural schemes based on topical diagnosis. All the symptoms were divided into blocks containing information about functional state of separate CNS structures and systems, character and degree of observed deviation. The analysis of obtained results permits to consider dysfunction of hypothalamus-hypophysis-adrenal system under emotional stress as a cause of stutter. Peculiarities of the neurohumoral regulatory system development determine both age and sex specificity of the considered syndrome. PMID- 10812673 TI - [Fibromyalgia (generalized tendomyopathy): defect of a program of movements and their realization]. AB - It was established that fulfillment of any motor action implies a design of the program of action in CNS and its realization by peripheral elements of locomotor system. A program includes an acceptor of the result of action and efferent synthesis on a central level (parietal-premotor area, pallidostriatal system, lymbico-reticular complex, ex cetera). A very motor act is realized by the activity of a segmental apparatus of the spinal cord and skeletal muscles. Acceptor of the result of action provides a control of the coincidence between afferent parameters of the result of an action and its real characteristics by means of the reverse feed-back (P.K. Anokhin, 1975). Incoordination of its parameters is a condition for the existence of a physiologic functional system for the construction of the necessary movement, while a coincidence of their indices resulted in the disintegration of this system. Appearance of myofascicular hypertonus in this system distorts a real parameters of the movement because of a deficit and imbalance of the proprioception in a segmental apparatus and central formations. A proprioceptive desafferentation promotes disinhibition of the neurons and formation of the generators of pathologically increased irritation with positive feed-back on rubro-segmental (A), pallido thalamic (B), strio-piramidal (C) and parietal-premotor (D) levels of the construction of the movement according to N.A. Bernshtein (1966). Its appearance on the B and C levels prevents destruction of the physiologic system of organization after realization of its activity and promotes its reformation into the pathologic one. It manifested clinically in appearance of a pathologic dynamic stereotype. Additional integration of lymbico-reticular complex, and high centers of autonomic supplement into this system promotes a formation of the stable neurotic, depressive reactions and autonomic disorders. Such reactions are transitory and have adaptive character at normal program of construction and fulfillment of the motion, while at pathologic one they have desadaptive character. Clinically they are known as multiple psychoautonomic syndromes combined with muscular, fascial and ligamental pains of different location. PMID- 10812674 TI - [Etiologic and pathogenetic factors of the disproportions in craniovertebral region]. PMID- 10812675 TI - [Phagocytic activity of neutrophils in patients with schizophrenia and schizoaffective psychosis]. PMID- 10812676 TI - [A case of delayed diagnosis of Wilson's disease]. PMID- 10812677 TI - [Fibromyalgia]. PMID- 10812678 TI - Anti-inflammatory triterpenoids from mysterious mushroom Ganoderma lucidum and their potential possibility in modern medicine. AB - Ganoderma lucidum, a mushroom long used in the East for a broad range of disorders, contains numerous pharmacologically active compounds. Very important of them are highly oxygenated anti-inflammatory triterpenes, which are the aim of this mini-review. PMID- 10812679 TI - Neuroprotective effects of antidotes in soman-poisoned rats. AB - 1. The neuroprotective effects of antidotes (atropine, obidoxime/atropine mixture, HI-6/atropine mixture) on rats poisoned with soman at a sublethal dose (48 micrograms/kg i.m.; 60% of LD50 value) were studied. The neurotoxicity was monitored using a functional observational battery (FOB) and an automatic measurement of motor activity. The neurotoxicity of soman was monitored at 24 h and 7 d following soman poisoning. 2. The results indicate that atropine alone and the oxime HI-6 in combination with atropine seem to be effective antidotal treatment for the elimination of soman-induced neurotoxicity in the case of sublethal poisonings. 3. On the other hand, the combination of obidoxime with atropine appears to be practically ineffective in diminishing neurotoxic soman induced symptoms. 4. Dealing with neuroprotective effects of antidotes, the oxime HI-6 in combination with atropine seems to be more suitable antidotal mixture than obidoxime in combination with atropine even in the case of sublethal poisoning with nerve agents. PMID- 10812680 TI - Are contrast sensitivity functions impaired in insulin dependent diabetics without diabetic retinopathy? AB - PURPOSE: To confirm the influence of multilevel metabolic disturbance of insulin dependent diabetes mellitus (IDDM) on the vision even before the onset of the other changes routinely evaluated by ophthalmologists. METHODS: Contrast sensitivity functions (CSFs) were estimated using the VCTS 6500 board. The standardised measurement procedure was performed. The value of the threshold contrast sensitivity was obtained for five spatial frequencies (1.5-3-6-12-18 c/deg). Other data was collected (duration of diabetes, BCVA, funduscopy, fluoresceine angiography, HbA1C). The study group consisted of 48 IDDM patients (94 eyes) without diabetic retinopathy and with Snellen BCVA > 1.0. The control group (56 normals, 98 eyes) was age and BCVA matched. RESULTS: Highly statistically significant decrease of the CSFs in all spatial frequencies in the study group was obtained. Correlation between duration of the diabetes and impaired degree of CSFs was present in the middle spatial frequency. No significant changes in CSFs were found among patients with pathological value of glycated hemoglobin HbA1c (> 7.8%). CONCLUSIONS: If compared with routinely used Snellen visual acuity, the CSFs are more complex descriptors of the subjects vision abilities. IDDM has an influence on these sensitive functions, especially during examination in the middle spatial frequency of 6 and 12 c/deg, before disturbing visual acuity and before changes in the retinal morphology. Decrease of CSFs was influenced mainly by the patients' age and partially (in the middle spatial frequency) by the IDDM duration. PMID- 10812681 TI - Topical use of aprotinin in coronary artery bypass surgery. AB - OBJECTIVE: The internal mammary artery (IMA) ranks among excellent, widely used conduits for surgical coronary revascularization. Its harvesting and its using may cause other surgical and technical problems and complications and increase postoperative bleeding from wound surface after the IMA harvesting with significantly greater incidence of blood transfusion. The aim of this study was to get to know how much it increases postoperative bleeding losses and if the local application of aprotinin (to the wound surface after the IMA harvesting and into the pericardial cavity) can reduce them and thus decrease the number of blood transfusions. METHODS: In this study there are compared groups of patients (n = 275) operated at the University Department of Cardiac Surgery in Hradec Kralove on account of ischemic heart disease. In the first part of this study results of operations of 200 patients were comprised retrospectively. Group A1 comprised 50 patients where for revascularization of the myocardium venous grafts were used. Group B1 comprised 50 patients where also the internal mammary artery was used. Group C1 was formed by 50 patients where after preparation of the IMA aprotinin (100,000 KIU) was administered locally to the wound surface after the IMA harvesting. Group D1 was formed by 50 patients where aprotinin (500,000 KIU) was administered locally to the wound surface and poured into the pericardial cavity before closure of the median sternotomy. The postoperative blood losses and the number of the administered blood transfusions were compared between these groups. RESULTS: The authors provided evidence that the using of the IMA increases significantly the postoperative blood losses (in group A1 675 ml +/- 352.9, in group B1 1232 ml +/- 336.5) and increases the number of required transfusions (in group A1 2.44 +/- 1.7, in group B1 3.45 +/- 1.0). By local aprotinin application to the wound surface after the IMA harvesting the blood losses and the number of administered transfusions were reduced in group C1 (896 ml (231.9, 2.74 +/- 0.8). In group D1 (local aprotinin application to the wound surface and into the pericardial cavity) the blood losses and the number of transfusions were increasingly reduced than in group C1 (797 ml +/- 280.5, 1.74 +/- 1.3). In the second, prospective randomised part of this study 3 groups of patients were compared. Group A2 comprised 25 patients where venous grafts for revascularization of myocardium were used. Group B2 was formed by 25 patients where also the IMA was used. Group D2 comprised 25 patients where aprotinin (500,000 u.) was administered locally to the wound surface after the IMA harvesting and poured into the pericardial cavity before closure of sternotomy. The postoperative blood losses and the number of administered blood transfusions were again compared between these groups. The total postoperative blood losses were 778 ml +/- 304.2 in group A2, 1072 ml +/- 391.8 in group B2 and 754 ml +/- 197.9 in group D2. There were compared blood losses after 6, 12 and 24 hours, too. There were the statistically significant differences among these groups during the whole postoperative period. The number of blood transfusions were 2.8 +/- 2.3 in group A2 and 2.04 +/- 1.1 in group B2. The use of aprotinin decreased this number in group D2, 1.44 +/- 1.1. CONCLUSIONS: The authors provided evidence that the harvesting and the using of the internal mammary artery for myocardial revascularization increases significantly the postoperative bleeding and increases the number of required transfusions. By local application of aprotinin the author reduced the blood losses and need of transfusions. PMID- 10812682 TI - Identification of monoclonal immunoglobulins by immunofixation electrophoresis- some questions. AB - In the series of 2413 paraproteins analyzed by immunoelectrophoresis during 30 years, doubled paraproteinemias were found 42 times, i.e. a frequence of 1.7%. In last two years we have found 202 paraproteins by means of immunofixation electrophoresis and within this group multiple paraproteinemias was found 21 times, i.e. a frequence of 10.4%. PMID- 10812683 TI - Less frequent neurologic complications of anticoagulant therapy. AB - One of the less frequent complications of anticoagulant therapy is damage to the femoral and sciatic nerves due to bleeding. This report presents two cases and discusses the pathogenetic mechanism and treatment of this medical problem. PMID- 10812684 TI - Infants with congenital deafness: on the importance of early sign language acquisition. AB - Two boys who both had a profound bilateral hearing impairment met at a specialized sign preschool. Their preconditions were quite different, since in one of them the hearing impairment was detected in the maternity ward with the aid of otoacoustic emissions, and habilitation had begun at age 4 months. The other boy's impairment was not detected until age 2 years; habilitation was thus much delayed. Data were collected on the two boys using interviews with parents and teachers, observation, and video recording in the children's own environment at home and in the specialized sign preschool. Characteristic differences between the boys are described regarding their social and linguistic development relating to the time of detection of the hearing impairment. This illustrates the importance of early detection and habilitation so as to avoid separation of individuals into different groups with differing social and academic prospects, depending on the lack of early linguistic stimulation and consequent poor language acquisition. Giving children the possibility of developing a language is the primary consideration. PMID- 10812685 TI - Deaf teacher candidates in hearing classrooms: a unique teacher preparation program. AB - An undergraduate teacher education program at Gallaudet University prepares deaf students in "regular" education. This includes a required full-time internship with hearing students (assisted by sign language interpreters). Graduates then continue in a master's degree program in deaf education, thus acquiring dual certification. Several studies indicate that these deaf candidates progress through the same developmental stages as hearing candidates and that they develop high expectations for deaf learners. Issues related to implementing such a program are discussed. PMID- 10812686 TI - Hand/wrist disorders among sign language communicators. AB - The study assessed the frequency of self-reported hand/wrist problems among sign language communicators, including interpreters, deaf or hard-of-hearing individuals, and educators. Comparisons were also made between sign language communicators and the general population regarding the frequency of medically diagnosed carpal tunnel syndrome. A survey was mailed to attendees of a national conference that focused on the quality of postsecondary educational opportunities for students with hearing deficits. Fifty-nine percent of 184 respondents reported experiencing hand/wrist problems. Twenty-six percent reported experiencing hand/wrist problems severe enough to limit the ability to work, and 18% reported a medical diagnosis of wrist tendinitis, carpal tunnel syndrome, or both. The frequency of self-reported, medically diagnosed carpal tunnel syndrome for the surveyed sign language communicators was five times the rate experienced by working Americans between ages 18 and 60 years (Tanaka, Wild, Seigman, Halperin, Behrens, & Putz-Anderson, 1995). The study supports increased emphasis on and availability of hand/wrist disorder risk reduction programs for sign language users. PMID- 10812687 TI - Cochlear implants for younger children: a preliminary description of the parental decision process and outcomes. AB - The authors conducted a preliminary telephone interview study of a random sample of 35 parents whose children had received cochlear implants through a large-scale implant program. Parents were asked about their child's preimplant and postimplant communications skills, how they learned about implants, and how they arrived at the decision to have their child receive an implant. Results of the interviews suggest, a least for this program, that two types of decision sequences are followed. One type of parent has initial and primary contact through a medical practitioner, uses that source of information exclusively, and is motivated by a desire for a "normal" communication situation. The second type of parent learns about implants from another parent, family member, or teacher. This individual will seek other sources of information and is most often motivated by the child's lack of communication skill. Generally, from parents' perspective, language and speech rather than improved social skills or social contact are the primary benefits of the implant. PMID- 10812688 TI - Helping our students meet the standards through test preparation classes. AB - Test preparation helps deaf students develop the skills they need to do well on standardized tests, particularly on a reading test format such as that used with the Stanford Achievement Test. The authors share information on how test preparation helps students to develop the skills required to do well on standardized reading tests by examining test-taking strategies and common language structures used in test items and how readers can learn to interpret them. The authors list the common errors made by their students and discuss remedial classes designed to address a range of test-taking and reading skills. PMID- 10812689 TI - Hearing impairment, social networks, and coping: the need for families with hearing-impaired children to relate to other parents and to hearing-impaired adults. AB - For a report on the stress experiences of parents with hearing-impaired children in Germany, 317 parents completed a survey on how their families communicate and socialize, among other issues. The report focuses on how contacts with other parents and with hearing-impaired adults affect stress experiences, in the context of the child's hearing status and the means of communication. Parents who frequently meet with other parents show evidence of a warm, accepting, trusting relationship with their child. Parents who have many contacts with hearing impaired adults show evidence of a strong sense of competence in regard to their child's upbringing. The findings confirm the implication found in most reports describing empirical studies. Social support is to be regarded as a cornerstone of psychosocial intervention and has to play as great a role as possible in institutional programs. PMID- 10812690 TI - Teachers' ratings of functional communication in students with cochlear implants. AB - The study examined factors associated with teachers' ratings of functional communication skills of students with cochlear implants. Deaf students living in and around a metropolitan area were surveyed to locate 51 with cochlear implants. Teachers rated each student's functional use of the implant, given three defined ratings. Additional information regarding sex, communication option, placement, home language, rural or nonrural address, etiology, and presence or absence of an additional disability was gathered. Chi-square analyses of the data were performed. The data indicated that students with a known etiology and a rural address, and who used sign language at home or school, were less likely than others to use the implant as a primary channel for receptive communication. The authors suggest that the teacher's role in implant use warrants more attention. Speech-language pathologists, otologists, audiologists, and parents of deaf children should consider all factors related to successful implant use before advocating or choosing this financially, emotionally, and therapeutically challenging option. PMID- 10812691 TI - Two pathways to prevention. AB - Health promotion and disease prevention programs are becoming important components of contemporary health care. There are at least 2 pathways to the enhancement of population health status through disease prevention. The first pathway requires the early diagnosis and treatment of disease. The second pathway promotes healthy lifestyles and disregards the requirement that a condition must be diagnosed before intervention is recommended. Data from several evaluations suggest that prevention efforts that rely on diagnosis have produced somewhat limited benefits, whereas primary prevention efforts may have substantial benefits. Current health policy places greater emphasis on secondary prevention. The objectives of improved population health might be better achieved by devoting relatively more resources to primary prevention through the promotion of healthy behaviors. PMID- 10812692 TI - Between the modern and the postmodern. The possibility of self and progressive understanding in psychology. AB - Psychology assumes defensible notions of human subjectivity and understanding. Yet, some versions of postmodernism, including some of those currently influential within psychology, eschew the possibility of the kind of understanding-capable personhood on which modern psychology has depended. The authors argue for a skeptical, middle-ground position that might allow psychologists to resist a forced choice between modernism and postmodernism in their subject matter and understanding. The authors set up their argument with 2 stories of human development and change. These stories assume no fixed, essentialist foundations of the sort favored in classically modern psychologies, yet they maintain the possibility of both self and understanding within a real but contingent physical, biological, and sociocultural world. The authors then articulate a middle-ground position as one that avoids the fixed foundationalism, essentialism, and absolute certitude of modernity, without endorsing the radical arbitrariness, antisubjectivism, and anarchistic relativism of some versions of postmodernity. PMID- 10812693 TI - The coming of age of telecommunications in psychological research and practice. AB - Rapid and far-reaching technological advances are revolutionizing the ways in which people relate, communicate, and live their daily lives. Technologies that were hardly used a few years ago, such as the Internet, e-mail, and video teleconferencing, are becoming familiar methods for modern communication. Telecommunications will continue to evolve quickly, spawning telehealth applications for research and the provision of clinical care in communities, university settings, clinics, and medical facilities. The impact on psychology will be significant. This article examines the application of developing technologies as they relate to psychology and discusses implications for professional research and practice. PMID- 10812694 TI - Conflicts and missed signals in psychoanalysis, behaviorism, and Gestalt psychology. AB - At the turn of the 20th century, European psychologists found themselves in conflict situations with respect to the role that private mental states should play in a scientific psychology. Out of this conflict arose 3 of the best-known schools of the 20th century: psychoanalysis, behaviorism, and Gestalt psychology. Each of these schools is discussed with respect to two characteristics. First, the authors discuss the attitude of each school on the meaning of the word unconscious as it was understood around 1900. Second, the authors discuss the influence of each school on the reception accorded to books written around 1900 espousing viewpoints that did not cohere with the school's beliefs. Such books may be considered "missed signals" in the history of psychology. "Hits" associated with each school are also highlighted. PMID- 10812695 TI - Social psychology a century ago. AB - America's dual interest in social welfare and practical science fueled the emergence of social psychology in the late 19th century. By that century's end, psychologists had presented diverse and sometimes contesting models of social psychology. These varied conceptualizations, however, were challenged by the discipline's growing dedication to the scientific method and experimentation, as well as the profession's need to produce knowledge that could be readily used to address current social problems and regulate social institutions. An appreciation of this late-19th-century moment of intellectual generativity and constraint affords an opportunity to reflect on the potentials and constraints of social psychology at the end of the 20th century. PMID- 10812696 TI - Psychologists in medical schools. PMID- 10812697 TI - 108th Convention of the American Psychological Association. Washington, DC, August 4-8, 2000 AB - Washington, DC--the nation's capital--celebrates a history rich in diversity and character. One of the most popular cities for sightseeing, Washington contains countless points of interest for its visitors. The world's largest museum complex, the Smithsonian Institution, invites you to explore exhibits that highlight the scientific, cultural, political, and technological developments of the United States and its people. Visit the home to original pieces of the American heritage, such as the Declaration of Independence, the Constitution, and the Bill of Rights, that have helped to shape the way we live today. The art lovers among you will delight in the seven major art galleries. See the modern art and sculpture of the Hirshhorn Museum and the newly opened Sculpture Garden; the Sackler Gallery's collection of Asian art; and the only museum devoted to the art and culture of Africa, the National Museum of African Art. In Washington, there is music in the air, from the Kennedy Center's many stages and the clubs of Georgetown and Adams Morgan to the military bands that give concerts on the Mall. Whatever your culinary desire, be it authentic Texas chili or the finest Asian cuisine, you'll find it at one of the city's internationally famous eateries. What a perfect place for APA to convene its first annual convention of the new millennium! PMID- 10812698 TI - Allergic contact dermatitis in dentistry. AB - Allergic contact dermatitis (ACD) in dentistry may affect dentists and orthodontists, technicians, nurses and patients. Changes to dental practice in recent years have altered the reported frequencies of allergens causing ACD in both dental personnel and patients. Allergic contact dermatitis to medicaments, metals and glutaraldehyde were previously common allergens in dentistry; however, widespread adoption of rubber gloves by staff has resulted in a significant increase in ACD to glove allergens in both dental staff and their patients, while affording protection against the traditional allergens. Both public concerns about potential toxicity of metals in oral restorations and a greater demand for cosmetic dentistry, have resulted in greater use of acrylics and resins by dental personnel, exposing them to highly allergenic materials. Dermatologists need to be aware of the newer allergenic materials used in dentistry in order to correctly manage skin diseases in this high-risk group. PMID- 10812699 TI - Lasers in skin resurfacing. AB - Laser skin resurfacing has revolutionized the approach to facial skin rejuvenation over the last decade. It has also added an approach to managing both atrophic and hypertrophic scars. This paper will review the basic principles of laser skin resurfacing, the different lasers used, the approach to treatment and potential complications of the procedure, followed by a discussion of future prospects in the field. PMID- 10812700 TI - Prevalence of delayed hypersensitivity to the European standard series in a self selected population. AB - There are limited reports of the prevalence of positive reactions in healthy adults to patch tests with standard allergens; there are no recent comprehensive studies from Australia. Healthy adult volunteers (n = 219) from the Western Australian community were patch tested using the European standard series of allergens. Seventy-seven (35%) reacted to at least one allergen, positive patch tests being most prevalent to nickel sulfate (20%), potassium dichromate (9%), cobalt chloride (6%) and fragrance mix (4%). Prevalence of positive patch tests to nickel and chromate was higher than that reported for another healthy population, which may stem from self-selection of volunteers or geographical differences, including extent of exposure to allergens. PMID- 10812701 TI - Human cases of cattle ringworm due to Trichophyton verrucosum in Victoria, Australia. AB - Between 1962 and 1994, 32 isolates of Trichophyton verrucosum from cases of tinea corporis, tinea faciei and tinea capitis were referred to the Mycology Reference Laboratory of the Microbiological Diagnostic Unit at The University of Melbourne, Victoria, Australia. Patients had presented at clinics in metropolitan Melbourne and country towns in the State of Victoria, Australia. They included dairy and cattle farmers, a slaughterman who worked in an abattoir, a veterinary tutor and children who lived on farms. Many patients lived in one of the three dairy farming areas of Victoria. A few lived in the outer suburbs of Melbourne. One cattle farmer came from south-east New South Wales. Specimens were not received for examination from animal contacts of the patients. The only patient from overseas was a boy from Lebanon. The literature recording human infections due to T. verrucosum in Australia is reviewed. PMID- 10812702 TI - Ulceration of the urethral meatus after simultaneous pancreas-kidney transplantation. AB - A 39-year-old male developed painful ulceration of the glans penis following simultaneous pancreas and kidney transplantation for end-stage renal failure complicating insulin-dependent diabetes mellitus. Infection was excluded. Diversion of the pancreatic secretions away from the urinary bladder into the bowel resulted in healing. PMID- 10812703 TI - Steatocystoma multiplex suppurativum: oral isotretinoin treatment combined with cryotherapy. AB - A 30-year-old man presented with numerous papules, nodules and inflamed cysts. The lesions were located all over the body, including the scalp, except the palms and soles. His mother and one sister had had similar but less extensive lesions. Histopathology of the biopsy specimens obtained from the anterior chest wall, axillae and the back region was consistent with steatocystoma multiplex (SM). A diagnosis of steatocystoma multiplex suppurativum was made. The inflamed lesions were treated with oral isotretinoin (1 mg/kg per daily) for 6 months. At the same time, cryotherapy was used for non-suppurating lesions smaller than 2 cm. When the patient was evaluated 6 months later, cosmetic results were good. No new lesions have appeared in the subsequent 12-month follow up. PMID- 10812704 TI - Cutaneous histiocytic lymphangitis: an unusual manifestation of rheumatoid arthritis. AB - Two cases are presented of unusual cutaneous lesions associated with rheumatoid arthritis in underlying joints. The lesions were evanescent, erythematous and violaceous partly macular and partly indurated plaques, with a livedo-like pattern of erythema at the edge in one case. Histological changes were identical in the two cases. The major features were dilated, dermal lymphatics containing aggregates of inflammatory cells, mainly histiocytes, with adjacent perivascular lymphoid aggregates. An appropriate name for this reaction would appear to be cutaneous histiocytic lymphangitis. PMID- 10812705 TI - Permanent alopecia following chemotherapy and bone marrow transplantation. AB - Alopecia can be a psychologically daunting prospect for people requiring cancer chemotherapy. Fortunately, most patients experience only temporary hair loss. We report the case of a 23-year-old woman with chronic myeloid leukaemia who developed permanent, near-total alopecia of her scalp, eyebrows, eyelashes, axillary and public hair following busulphan and cyclophosphamide chemotherapy which was used as conditioning prior to allogeneic bone marrow transplantation. The histology from a scalp biopsy revealed hair follicle destruction. Topical minoxidil failed to induce significant re-growth. PMID- 10812706 TI - Benign melanocytic proliferative nodule within a congenital naevus. AB - A neonate presented with a deeply pigmented papule within a medium-sized congenital naevus. Histologically, this proved to be a benign proliferative nodule in a congenital naevus. This case is presented to highlight the occurrence of this lesion, the main differential of which is the rare entity of true congenital melanoma. PMID- 10812707 TI - Human immunodeficiency virus-related primary cutaneous aspergillosis. AB - A 31-year-old Caucasian man with AIDS developed a crusted violaceous plaque under adhesive tape near a central venous catheter insertion site. Histological examination demonstrated a ruptured hair follicle containing collections of fungal hyphae typical of Aspergillus spp. A culture of the biopsy material grew Aspergillus fumigatus. The patient responded to removal of the catheter and the occlusive dressing, in addition to itraconazole therapy. Aspergillosis must be considered in the differential diagnosis of cutaneous lesions in human immunodeficiency virus-infected patients, in particular when the lesion occurs under adhesive tape or an occlusive dressing. PMID- 10812708 TI - Perineural spread in head and neck skin cancer. AB - Perineural spread (PNS) in the head and neck is an infrequent but aggressive manifestation of skin cancer. As such, it can provide access to the intracranial cavity. Squamous cell carcinoma is the most common histology with the facial and trigeminal nerves most often involved. Orbital invasion is an uncommon but devastating result of PNS located around the orbit, particularly the forehead. Diagnosis can be difficult and initial investigations are often unhelpful. Treatment should ideally be directed at preventing further spread before it develops. Adjuvant radiotherapy is often recommended. The disease may present at an advanced state within the orbit or parotid gland or even within the intracranial cavity. Clinicians need to be aware of the potential for PNS because a group of these patients will die from this potentially preventable and treatable form of metastatic skin cancer. PMID- 10812709 TI - Folliculotropic T-cell lymphocytosis (mucin-poor follicular mucinosis). AB - A 48-year-old man presented with multiple asymptomatic patches of hair loss over his trunk and limbs associated with focal keratotic follicular plugs. Multiple skin biopsies showed a panfollicular lymphocytic infiltrate associated with follicular hyperkeratinization, minimal follicular spongiosis, focal basaloid follicular hyperplasia but no overt follicular mucinosis. The lymphocytes were small and there was no atypia. Immunoperoxidase stains showed that the follicular lymphocytes were T cells and predominantly CD4 positive with HLADr (LN3) expressed on their surface. There were insufficient clinical or histopathological features to make a diagnosis of folliculotropic T-cell lymphoma. This case currently may be classified best as folliculotropic T-cell lymphocytosis and may represent a mucin-poor counterpart of follicular mucinosis. Such cases may pursue an indolent course or may evolve to folliculotropic T-cell lymphoma, mycosis fungoides or anaplastic lymphoma. The term folliculotropic T-cell lymphocytosis may be useful for similar cases lacking clinical or histological criteria for lymphoma and lacking follicular mucinosis. PMID- 10812710 TI - Dermatitis following systemic prednisolone: patch testing with prednisolone eye drops. AB - Although positive reactions to topical corticosteroids can occur in over 1% of a patch-test population, systemic reactions to oral corticosteroids are uncommon. A 45-year-old woman who gave a clear history of the generalization of a cutaneous eruption following oral prednisolone was positive on patch testing using prednisolone eye drops. PMID- 10812711 TI - Kaposi's varicelliform eruption or atypical chickenpox in a normal individual. PMID- 10812712 TI - Clinical dermatology update: international meeting of the International League of Dermatological Societies. PMID- 10812713 TI - Clinical cutaneous allergy and immunology, an interface between two specialities. PMID- 10812714 TI - Population dependent Fourier decomposition of fitness landscapes over recombination spaces: evolvability of complex characters. AB - The effect of recombination on genotypes can be represented in the form of P structures, i.e., a map from the set of pairs of genotypes to the power set of genotypes. The interpretation is that the P-structure maps the pair of parental genotypes to the set of recombinant genotypes which result from the recombination of the parental genotypes. A recombination fitness landscape is then a function from the genotypes in a P-structure to the real numbers. In previous papers we have shown that the eigenfunctions of (a matrix associated with) the P-structure provide a basis for the Fourier decomposition of arbitrary recombination landscapes. Here we generalize this framework to include the effect of genotype frequencies, assuming linkage equilibrium. We find that the autocorrelation of the eigenfunctions of the population-weighted P-structure is independent of the population composition. As a consequence we can directly compare the performance of mutation and recombination operators by comparing the autocorrelations on the finite set of elementary landscapes. This comparison suggests that point mutation is a superior search strategy on landscapes with a low order and a moderate order of interaction p < n/3 (n is the number of loci). For more complex landscapes 1 point recombination is superior to both mutation and uniform recombination, but only if the distance among the interacting loci (defining length) is minimal. Furthermore we find that the autocorrelation on any landscape is increasing as the distribution of genotypes becomes more extreme, i.e., if the population occupies a location close to the boundary of the frequency simplex. Landscapes are smoother the more biased the distribution of genotype frequencies is. We suggest that this result explains the paradox that there is little epistatic interaction for quantitative traits detected in natural populations if one uses variance decomposition methods while there is evidence for strong interactions in molecular mapping studies for quantitative trait loci. PMID- 10812715 TI - Wavelengths of gyrotactic plumes in bioconvection. AB - Bioconvection occurs as the result of the collective behaviour of many micro organisms swimming in a fluid and is realized as patterns similar to those of thermal convection which occur when a layer of fluid is heated from below. We consider the phenomenon of pattern formation due to gyrotaxis, an orientation mechanism which results from the balance of gravitational and viscous torques acting on bottom-heavy micro-organisms. The continuum model of Pedley et al. (1988, J. Fluid. Mech. 195, 223-237) is used to describe the suspension. The system is governed by the Navier-Stokes equations for an incompressible fluid coupled with a micro-organism conservation equation. These equations are solved numerically using a conservative finite-difference scheme. To examine the dependence of the horizontal pattern wavelengths on the parameters, we consider two-dimensional solutions in a wide chamber using rigid side walls. The wavelengths of the numerical computations are in good agreement with the experimental observations and we provide the first computational examples of the commonly seen 'bottom-standing' plumes. PMID- 10812716 TI - Evolution in knockout conflicts: the fixed strategy case. AB - A group of individuals resolve their disputes by a knockout tournament. In each round of the tournament, the remaining contestants form pairs which compete, the winners progressing to the next round and the losers being eliminated. The payoff received depends upon how far the player has progressed and a cost is incurred only when it is defeated. We only consider strategies in which individuals are constrained to adopt a fixed play throughout the successive rounds. The case where individuals can vary their choice of behaviour from round to round will be treated elsewhere. The complexity of the system is investigated and illustrated both by special cases and numerical examples. PMID- 10812717 TI - Integrate-and-fire models with nonlinear leakage. AB - Can we express biophysical neuronal models as integrate-and-fire (IF) models with leakage coefficients which are no longer constant, as in the conventional leaky IF model, but functions of membrane potential and other biophysical variables? We illustrate the answer to this question using the FitzHugh-Nagumo (FHN) model as an example. A novel IF type model, the IF-FHN model, which approximates to the FHN model, is obtained. The leakage coefficient derived in the IF-FHN model has nonmonotonic relationship with membrane potential, revealing at least in part the intrinsic mechanisms underlying the FHN models. The IF-FHN model correspondingly exhibits more complex behaviour than the standard IF model. For example, in some parameter regions, the IF-FHN model has a coefficient of variation of the output interspike interval which is independent of the number of inhibitory inputs, being close to unity over the whole range, comparable to the FHN model as we noted previously (Brown et al., 1999). PMID- 10812718 TI - Enzyme kinetics at high enzyme concentration. AB - We re-visit previous analyses of the classical Michaelis-Menten substrate-enzyme reaction and, with the aid of the reverse quasi-steady-state assumption, we challenge the approximation d[C]/dt approximately 0 for the basic enzyme reaction at high enzyme concentration. For the first time, an approximate solution for the concentrations of the reactants uniformly valid in time is reported. Numerical simulations are presented to verify this solution. We show that an analytical approximation can be found for the reactants for each initial condition using the appropriate quasi-steady-state assumption. An advantage of the present formalism is that it provides a new procedure for fitting experimental data to determine reaction constants. Finally, a new necessary criterion is found that ensures the validity of the reverse quasi-steady-state assumption. This is verified numerically. PMID- 10812719 TI - A chemotactic model for the advance and retreat of the primitive streak in avian development. AB - The formation of the primitive streak in early avian development marks the onset of gastrulation, during which large scale cell movement leads to a trilaminar blastoderm comprising prospective endodermal, mesodermal and ectodermal tissue. During streak formation a specialized group of cells first moves anteriorly as a coherent column, beginning from the posterior end of the prospective anterior posterior axis (a process called progression), and then reverses course and returns to the most posterior point on the axis (a process called regression). To date little is known concerning the mechanisms controlling either progression or regression. Here we develop a model in which chemotaxis directs the cell movement and which is capable of reproducing the principal features connected with progression and regression of the primitive streak. We show that this model exhibits a number of experimentally-observed features of normal and abnormal streak development, and we propose a number of experimental tests which may serve to illuminate the mechanisms. This paper represents the first attempt to model the global features of primitive streak formation, and provides an initial stage in the development of a more biologically-realistic discrete cell model that will allow for variation of properties between cells and control over movement of individual cells. PMID- 10812720 TI - Diverse ideas on the growth kinetics of disseminated cancer cells. AB - Theoretical and empirical arguments are used to support the growth kinetics of disseminated tumors. Employing viable hypotheses, it is established that Gompertzian growth of disseminated cancer cells can be derived from a number of theoretical considerations. Empirical methods are used to validate and confirm the theoretical assertions with the use of available data. PMID- 10812721 TI - Multipole approach to the inverse problem in electrocardiology: convergence of the multipole equivalent generator on the inhomogeneous body conductor. AB - The multipole approach to the inverse electrocardiological problem consists of estimating the multipole components of the cardiac electric generator, starting from the measured body surface potential. This paper presents a critical investigation of the basic premise for the applicability of the multipole approach, namely the convergence of the multipole equivalent generator for the heart on the surface of an inhomogeneous body conductor. As an extension to multipole theory, a criterion for the convergence is derived. Based on realistic models for the body conductor and the cardiac electric generator, we observe that the criterion is not strictly satisfied in realistic conditions. Numerical simulations with the same models point out that the multipole equivalent generator is indeed not convergent in the strict mathematical sense. On the other hand, we show that the multipole equivalent generator yields a rather close approximation of the electrocardiological potential for intermediate values of the order of the multipole generator. A discussion is given on how to explain the apparently ambiguous results for the estimation of cardiac multipole components. PMID- 10812722 TI - Polio, war and peace. PMID- 10812723 TI - The legacies of polio eradication. PMID- 10812725 TI - Surveillance of patients with acute flaccid paralysis in Finland: report of a pilot study. AB - WHO recommends that surveillance of patients with acute flaccid paralysis (AFP) be used to demonstrate the eradication of wild poliovirus. In this article we report the results of a study to assess the frequency of AFP patients referred to Finnish hospitals and whether virological diagnostic coverage could be improved by repeated reminders and active feedback. For this purpose, we sent monthly questionnaires to all neurological and paediatric neurological units in Finland, requesting retrospective reporting on investigated paralytic patients with defined clinically relevant diagnoses, rather than AFP. Reminder letters included a pre-paid return envelope. Virological investigations were offered cost free. Of the 492 reporting forms sent, 415 (84%) were returned, evenly covering both the population and the study period (July 1997 to June 1998). Of the 90 patients reported, 83 were evaluable. The apparent incidences of the diagnoses covered were 1.6 per 100,000 at any age, and 1.0 per 100,000 for under--15-year-olds. Guillain-Barre syndrome was the most common diagnosis (0.80 per 100,000). The two faecal specimens required were virologically investigated in nine out of the 10 patients under 15 years of age, but in only 46% of all patients. Four adenovirus strains, but no polioviruses or other enteroviruses, were isolated. We conclude that a satisfactory monthly reporting system was readily established and that a sufficient number of patients with diagnoses resembling AFP are being referred to Finnish hospitals. Active feedback did not increase the proportion of virologically investigated patients to an acceptable level in all age groups. It is clear that other approaches must be used to quantify the circulation of poliovirus in Finland. PMID- 10812724 TI - Disease eradication as a public health strategy: a case study of poliomyelitis eradication. AB - Disease eradication as a public health strategy was discussed at international meetings in 1997 and 1998. In this article, the ongoing poliomyelitis eradication initiative is examined using the criteria for evaluating candidate diseases for eradication proposed at these meetings, which covered costs and benefits, biological determinants of eradicability (technical feasibility) and societal and political considerations (operational feasibility). The benefits of poliomyelitis eradication are shown to include a substantial investment in health services delivery, the elimination of a major cause of disability, and far-reaching intangible effects, such as establishment of a "culture of prevention". The costs are found to be financial and finite, despite some disturbances to the delivery of other health services. The "technical" feasibility of poliomyelitis eradication is seen in the absence of a non-human reservoir and the presence of both an effective intervention and delivery strategy (oral poliovirus vaccine and national immunization days) and a sensitive and specific diagnostic tool (viral culture of specimens from acute flaccid paralysis cases). The certification of poliomyelitis eradication in the Americas in 1994 and interruption of endemic transmission in the Western Pacific since March 1997 confirm the operational feasibility of this goal. When the humanitarian, economic and consequent benefits of this initiative are measured against the costs, a strong argument is made for eradication as a valuable disease control strategy. PMID- 10812726 TI - Polio as a platform: using national immunization days to deliver vitamin A supplements. AB - In 1988 the 41st World Health Assembly committed WHO to the goal of global eradication of poliomyelitis by 2000 "in ways which strengthen national immunization programmes and health infrastructure". The successful use of polio National Immunization Days (NIDs) to deliver vitamin A is an example of how polio eradication can serve as a platform to address other problems of child health. Importantly, this integration is helping to achieve the World Summit for Children goal of eliminating vitamin A deficiency by the year 2000. It is estimated that between 140 million and 250 million preschool children are at risk of subclinical vitamin A deficiency. In 1998 more than 60 million children at risk received vitamin A supplements during polio national immunization days (NIDs). While food fortification and dietary approaches are fundamental to combating vitamin A deficiency, the administration of vitamin A supplements during NIDs helps raise awareness, enhance technical capacity, improve assessment and establish a reporting system. Moreover, polio NIDs provide an entry point for the sustainable provision of vitamin A supplements with routine immunization services and demonstrate how immunization campaigns can be used for the delivery of other preventive health services. PMID- 10812727 TI - Excluding polio in areas of inadequate surveillance in the final stages of eradication in China. AB - In 1996, China adopted a virological classification of acute flaccid paralysis (AFP) cases for its surveillance system. Only AFP cases with wild poliovirus in stool specimens are confirmed as polio. Cases with adequate stool specimens that are negative for wild poliovirus are not counted. This paper describes a methodology to rule out poliomyelitis in AFP cases with inadequate stool specimens. National surveillance data were analysed using dot maps to detect clusters of AFP cases with high-risk factors for poliomyelitis. The surveillance system and vaccine coverage were assessed during field investigations. Four clusters of AFP cases were identified, but no poliomyelitis cases. Programmatic failures in the identified high-risk areas included low vaccination rates, poor stool specimen collection and inadequate AFP surveillance. Programme strategies were implemented to correct the identified failures. Use of this methodology provides strong evidence consistent with the absence of wild poliovirus in China. PMID- 10812728 TI - Poliomyelitis surveillance: the model used in India for polio eradication. AB - Poliomyelitis surveillance in India previously involved the passive reporting of clinically suspected cases. The capacity for detecting the disease was limited because there was no surveillance of acute flaccid paralysis (AFP). In October 1997, 59 specially trained Surveillance Medical Officers were deployed throughout the country to establish active AFP surveillance; 11,533 units were created to report weekly on the occurrence of AFP cases at the district, state and national levels; timely case investigation and the collection of stool specimens from AFP cases was undertaken; linkages were made to support the polio laboratory network; and extensive training of government counterparts of the Surveillance Medical Officers was conducted. Data reported at the national level are analysed and distributed weekly. Annualized rates of non-polio AFP increased from 0.22 per 100,000 children aged under 15 years in 1997 to 1.39 per 100,000 in 1999. The proportion of cases with two adequate stools collected within two weeks of the onset of paralysis increased from 34% in 1997 to 68% in 1999. The number of polio cases associated with the isolation of wild poliovirus decreased from 211 in the first quarter of 1998 to 77 in the first quarter of 1999. Widespread transmission of wild poliovirus types 1 and 3 persists throughout the country; type 2 occurs only in Bihar and Uttar Pradesh. In order to achieve polio eradication in India during 2000, extra national immunization days and house-to-house mopping-up rounds should be organized. PMID- 10812729 TI - Eradication of poliomyelitis in countries affected by conflict. AB - The global initiative to eradicate poliomyelitis is focusing on a small number of countries in Africa (Angola, Democratic Republic of the Congo, Liberia, Sierra Leone, Somalia, Sudan) and Asia (Afghanistan, Tajikistan), where progress has been hindered by armed conflict. In these countries the disintegration of health systems and difficulties of access are major obstacles to the immunization and surveillance strategies necessary for polio eradication. In such circumstances, eradication requires special endeavours, such as the negotiation of ceasefires and truces and the winning of increased direct involvement by communities. Transmission of poliovirus was interrupted during conflicts in Cambodia, Colombia, El Salvador, Peru, the Philippines, and Sri Lanka. Efforts to achieve eradication in areas of conflict have led to extra health benefits: equity in access to immunization, brought about because every child has to be reached; the revitalization and strengthening of routine immunization services through additional externally provided resources; and the establishment of disease surveillance systems. The goal of polio eradication by the end of 2000 remains attainable if supplementary immunization and surveillance can be accelerated in countries affected by conflict. PMID- 10812730 TI - Massive outbreak of poliomyelitis caused by type-3 wild poliovirus in Angola in 1999. AB - The largest outbreak of poliomyelitis ever recorded in Africa (1093 cases) occurred from 1 March to 28 May 1999 in Luanda, Angola, and in surrounding areas. The outbreak was caused primarily by a type-3 wild poliovirus, although type-1 wild poliovirus was circulating in the outbreak area at the same time. Infected individuals ranged in age from 2 months to 22 years; 788 individuals (72%) were younger than 3 years. Of the 590 individuals whose vaccination status was known, 23% had received no vaccine and 54% had received fewer than three doses of oral poliovirus vaccine (OPV). The major factors that contributed to this outbreak were as follows: massive displacement of unvaccinated persons to urban settings; low routine OPV coverage; inaccessible populations during the previous three national immunization days (NIDs); and inadequate sanitation. This outbreak indicates the urgent need to improve accessibility to all children during NIDs and the dramatic impact that war can have by displacing persons and impeding access to routine immunizations. The period immediately after an outbreak provides an enhanced opportunity to eradicate poliomyelitis. If continuous access in all districts for acute flaccid paralysis surveillance and supplemental immunizations cannot be assured, the current war in Angola may threaten global poliomyelitis eradication. PMID- 10812731 TI - Stopping poliovirus vaccination after eradication: issues and challenges. AB - Since 1988 reported polio cases worldwide have declined by about 85% and the number of known or suspected polioendemic countries has decreased from over 120 to less than 50. With eradication of poliomyelitis approaching, issues potentially affecting when and how vaccination against poliovirus can be stopped become extremely important. Because of the potential risks and benefits inherent in such a decision, the best available science, a risk-benefit analysis, contingency plans, a stock pile of poliovirus vaccines, and the endorsement by the global policy-making committees will all be needed before vaccination can be discontinued. The scientific basis for stopping polio immunization has been reviewed by WHO. This Round Table article summarizes the current state of knowledge, provides an update on the processes and timelines for certification, containment, and stopping vaccination, and highlights some of the unanswered scientific questions that will be addressed by further research. These include whether transmission of vaccine-derived poliovirus strains could be sustained so that poliomyelitis could re-emerge in a future unvaccinated population and whether prolonged excretion of vaccine-derived poliovirus from individuals with immune deficiencies could be a mechanism through which this could occur. PMID- 10812732 TI - Gaps in our knowledge about transmission of vaccine-derived polioviruses. PMID- 10812733 TI - It is too early to stop polio vaccination. PMID- 10812734 TI - The risks of stopping vaccination: perspectives from the developing world. PMID- 10812735 TI - What laboratory studies will reduce the risk of wild poliovirus reservoirs being missed? PMID- 10812736 TI - Elevated levels of maternal anti-tetanus toxin antibodies do not suppress the immune response to a Haemophilus influenzae type b polyribosylphosphate-tetanus toxoid conjugate vaccine. AB - Reported are the effects of elevated levels of anti-tetanus antibodies on the safety and immune response to a Haemophilus influenzae type b polyribosylphosphate (PRP)-tetanus toxoid conjugate (PRP-T) vaccine. A group of Thai infants (n = 177) born to women immunized against tetanus during pregnancy were vaccinated with either a combined diphtheria-tetanus-pertussis (DTP) PRP-T vaccine or DTP and a PRP-conjugate vaccine using Neisseria meningitidis group B outer-membrane proteins as a carrier (PedVax HIB). Although most infants possessed high titres (> 1 IU/ml) of anti-tetanus antibodies, the DTP-PRP-T combined vaccine engendered an excellent antibody response to all vaccine components. In both vaccine groups > 98% of infants attained anti-PRP antibody titres > or = 0.15 microgram/ml. The geometric mean anti-PRP antibody titres were 5.41 micrograms/ml and 2.1 micrograms/ml for infants immunized with three doses of PRP-T versus two doses of PedVax HIB vaccines, respectively (P < 0.005). Similarly, the proportion of infants who achieved titres > or = 1 microgram/ml was higher in the PRP-T group (87.8%) than in the group immunized with PedVax HIB (74.2%) (P = 0.036). A subgroup analysis showed that there was no significant difference in the anti-PRP antibody response for infants exhibiting either < 1 IU of anti-tetanus antibody per millilitre or > or = 1 IU/ml at baseline. These finding indicate that pre-existing anti-carrier antibody does not diminish the immune response to the PRP moiety. All infants possessed protective levels of anti-D and anti-T antibody levels after immunization. PMID- 10812737 TI - Randomized controlled trial of anterior-chamber intraocular lenses in Nepal: long term follow-up. AB - Most of the estimated 20 million people who are blind with cataracts live in rural areas of developing countries, where expert surgical resources are scarce. We have studied the use of multiflex open-loop anterior-chamber intraocular lenses (ACIOL) in high-volume low-cost surgery. Between 1992 and 1995, a total of 2000 people attending Lahan Eye Hospital, Nepal, with bilateral cataracts reducing vision to < or = 6/36 were randomly allocated to receive intracapsular extraction (ICCE) with aphakic spectacles, or ICCE with an ACIOL. We re-examined the cohort (1305/2000, 65%) between November 1996 and April 1997 and report the findings in this article. There were 13 new cases of poor visual outcome (best corrected vision < 6/60) arising after one year: 9 in the ACIOL group and 4 in the control group; odds ratio 2.1 (95% confidence interval, 0.59-9.55). The causes of poor outcome were as follows: ACIOL group--retinal detachment (4 cases), cystoid macular oedema (2), epiretinal membrane (1), age-related macular degeneration (1), and late endophthalmitis (1); control group--retinal detachment (2 cases), late endophthalmitis (1), and primary open-angle glaucoma with age related macular degeneration (1). In rural areas of developing countries, well manufactured multiflex open-loop ACIOLs can be implanted safely by experienced ophthalmologists after routine ICCE, avoiding the disadvantages of aphakic spectacle correction. PMID- 10812738 TI - The role of son preference in reproductive behaviour in Pakistan. AB - The sex of surviving children is an important determinant of reproductive behaviour in South Asia in general and Pakistan in particular. This cohort study evaluates the role of the sex of children on reproductive intentions and subsequent behaviour of women in urban slums of Karachi, Pakistan. The analysis is based on two rounds of surveys conducted in 1990-91 and 1995 of a cohort of married women aged 15-49 years. The results show that pregnancies became increasingly unwanted as the number of surviving sons increased. The sex of surviving children was strongly correlated with subsequent fertility and contraceptive behaviour. However, rather than an exclusive son preference, couples strove for one or more sons and at least one surviving daughter. The policy implications of the link between overt son preference and low status of women are discussed. PMID- 10812739 TI - Rapid screening for Schistosoma mansoni in western Cote d'Ivoire using a simple school questionnaire. AB - The distribution of schistosomiasis is focal, so if the resources available for control are to be used most effectively, they need to be directed towards the individuals and/or communities at highest risk of morbidity from schistosomiasis. Rapid and inexpensive ways of doing this are needed, such as simple school questionnaires. The present study used such questionnaires in an area of western Cote d'Ivoire where Schistosoma mansoni is endemic; correctly completed questionnaires were returned from 121 out of 134 schools (90.3%), with 12,227 children interviewed individually. The presence of S. mansoni was verified by microscopic examination in 60 randomly selected schools, where 5047 schoolchildren provided two consecutive stool samples for Kato-Katz thick smears. For all samples it was found that 54.4% of individuals were infected with S. mansoni. Moreover, individuals infected with S. mansoni reported "bloody diarrhoea", "blood in stools" and "schistosomiasis" significantly more often than uninfected children. At the school level, Spearman rank correlation analysis showed that the prevalence of S. mansoni significantly correlated with the prevalence of reported bloody diarrhoea (P = 0.002), reported blood in stools (P = 0.014) and reported schistosomiasis (P = 0.011). Reported bloody diarrhoea and reported blood in stools had the best diagnostic performance (sensitivity: 88.2%, specificity: 57.7%, positive predictive value: 73.2%, negative predictive value: 78.9%). The study, which is probably the largest of its kind ever undertaken in Africa, revealed a moderate diagnostic performance of questionnaires for identifying individuals and/or communities at high risk from S. mansoni. PMID- 10812741 TI - Malaria drug resistance. PMID- 10812740 TI - A proposal to declare neurocysticercosis an international reportable disease. AB - Neurocysticercosis is an infection of the nervous system caused by Taenia solium. It is the most important human parasitic neurological disease and a common cause of epilepsy in Africa, Asia, and Latin America, representing enormous costs for anticonvulsants, medical resources and lost production. Neurocysticercosis is a human-to-human infection, acquired by the faecal-enteric route from carriers of intestinal T. solium, most often in areas with deficient sanitation. Intestinal tapeworms cause few symptoms, but adult taeniae carried by humans release large numbers of infective eggs and are extremely contagious. Ingestion of poorly cooked pig meat infested with T. solium larvae results in intestinal taeniosis but not neurocysticercosis. With a view to hastening the control of taeniosis and neurocysticercosis we propose that neurocysticercosis be declared an international reportable disease. New cases of neurocysticercosis should be reported by physicians or hospital administrators to their health ministries. An epidemiological intervention could then be launched to interrupt the chain of transmission by: (1) searching for, treating and reporting the sources of contagion, i.e. human carriers of tapeworms; (2) identifying and treating other exposed contacts; (3) providing health education on parasite transmission and improvement of hygiene and sanitary conditions; and (4) enforcing meat inspection policies and limiting the animal reservoir by treatment of pigs. We believe that the first step required to solve the problem of neurocysticercosis is to implement appropriate surveillance mechanisms under the responsibility of ministries of health. Compulsory notification also has the major advantage of providing accurate quantification of the incidence and prevalence of neurocysticercosis at regional level, thus permitting the rational use of resources in eradication campaigns. PMID- 10812742 TI - Community care not to blame for increased offending among the mentally ill. PMID- 10812743 TI - More partnerships to spur vaccine development. PMID- 10812744 TI - Community-based health care and development: exploring the myths. PMID- 10812745 TI - Outbreak of salmonellosis associated with chicks and ducklings at a children's nursery. PMID- 10812746 TI - Meningococcal infection in pilgrims returning from the Haj: update. PMID- 10812747 TI - Guidelines for preventing catheter associated urinary tract infections--on Internet for consultation. PMID- 10812748 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 10812749 TI - Presumptive summer influenza A: an outbreak on a trans-Tasman cruise. AB - A number of recent reports from the Northern Hemisphere have drawn attention to the occurrence of summer outbreaks (May to August) of influenza A among cruise ship passengers and their contacts. In cases amongst passengers returning to Canada from Alaska, exposure appears to have occurred during the land-based Alaskan tour with illness developing during the subsequent cruise. A late summer outbreak of influenza A among passengers and crew on the return leg of a 14-day Sydney-New Zealand-Sydney cruise is reported in this article. PMID- 10812750 TI - Possible community immunity to small round structured virus gastroenteritis in a rural aboriginal community. AB - In April 1998 an outbreak of gastroenteritis affected visitors, but none of the Aboriginal residents, at a Territory Health Services luncheon in a rural Aboriginal community in Central Australia. The epidemiological features and identification of Small Round Structured Virus (SRSV) from two participants suggest that this was an outbreak caused by a SRSV. The attack rate in the visitors who ate or drank food at the luncheon was 73% (11 of 15). Seventeen Aboriginal residents were interviewed, none had gastroenteritis. The community potable water supply was contaminated with faecal bacteria around the time of the outbreak. No particular food could be implicated and laboratory examination of foods was not possible. It is proposed that past exposure to SRSVs may have resulted in the Aboriginal residents developing clinical immunity to infection. The process and consequences of the investigation in this community are also discussed. PMID- 10812751 TI - Factors influencing vaccination uptake. Workshop report. Current Australian research on the behavioural, social and demographic factors influencing immunisation, Royal Alexandra Hospital for Children, Sydney, March 1998. AB - Current Australian research on factors influencing vaccination was discussed at a workshop held at the Royal Alexandra Hospital for Children, Sydney, in March 1998, sponsored by the National Centre for Immunisation Research and Surveillance of Vaccine Preventable Diseases (NCIRS). The application of decision making theory to vaccination behaviour, the expectations and experiences of mothers, and reasons why parents fail to vaccinate their children were considered. Mothers' perceptions of the risks of vaccines, preferences of parents and providers for the mode of vaccine delivery, and community and social factors were all found to be part of the framework within which vaccination is accepted in Australia. Consumer considerations, media influences and overseas comparisons were discussed. PMID- 10812752 TI - Communicable diseases surveillance. PMID- 10812753 TI - The effect of foot-bath with or without the essential oil of lavender on the autonomic nervous system: a randomized trial. AB - OBJECTIVES: This study was designed to investigate the effect of foot-bath with or without the essential oil of lavender on the autonomic nervous system. DESIGN: Randomized crossover controlled study. SETTING: Nursing college, Nagano, Japan. INTERVENTION: Young women sat with their feet soaked in hot water for 10 minutes with and without the essential oil. OUTCOME MEASURES: An electrocardiogram, finger tip blood flow and respiratory rate were recorded. Autonomic function was evaluated using spectral analysis of heart rate variability. RESULTS: The foot bath caused no changes in heart or respiratory rates, but produced a significant increase in blood flow. Using spectral analysis, the parasympathetic nerve activity increased significantly during the both types of foot-bath. In the case of the foot-bath with the addition of essential oil of lavender, there were delayed changes to the balance of autonomic activity in the direction associated with relaxation. CONCLUSION: A hot foot-bath and oil of lavender appear to be associated with small but significant changes in autonomic activity. PMID- 10812754 TI - Communication between general and manipulative practitioners: a survey. AB - A survey of general practitioners (GPs) in the south of England was undertaken to determine their understanding and communication needs in referring patients to practitioners of manipulation. Eighty-six out of 309 GPs replied to a postal questionnaire (28% response). The results suggest that, while routine communication is important for improving understanding, GPs appear to have a preference for disciplines of which they have personal experience. The majority of responders favoured receiving a report on one side of A5 paper when the patient completes treatment. This should contain the nature of treatment and advice given and an indication of its outcome. Those who desired an initial report wanted it to contain a summary of the nature of the problem, a brief history, a summary of relevant findings from the examination, any investigations and a prognosis. Many GPs commented that they were more comfortable in referring to physiotherapists because they felt they had a better understanding of the treatment involved. Furthermore, chiropractic and osteopathic terminologies were reported to be confusing more often than physiotherapy terminology. Bearing in mind the potential bias in responses due to its geographical limitations and low response rate, this study provides useful indicators for manipulative and GPs who wish to work more closely together. PMID- 10812755 TI - Considering the alternatives: a special study module in complementary therapy. AB - OBJECTIVES: This study sought to describe the motivation, experience and attitude change of 21 undergraduate medical students who chose a special study module in complementary therapy (CT). DESIGN: Survey; self-administered questionnaire before and after the module. SETTING: Second year medical students' course special study module in CT concentrating on homeopathy at The University of Birmingham Medical School, UK. RESULTS: Twenty questionnaires were completed at each time point with 19 sets of paired data. Students saw the module as offering a rare opportunity to examine and evaluate non-orthodox therapies. Eleven students had personal experience of a CT prior to the module, but there was a wide range of student attitudes from 'scepticism' to 'acceptance'. Students tended to rate the efficacy of CT as a whole, and homeopathy in isolation, similarly (P = 0.005). This trend was less marked at the end (P = 0.077). On both occasions CT was rated higher. Individual student's ratings moved in both directions. Twelve students considered learning and practising a CT, most commonly acupuncture. CONCLUSIONS: The evolution of UK undergraduate medical curricula has begun to incorporate non-core components through special study modules. The Birmingham CT module was popular and attracted students with a range of initial attitudes. We have demonstrated that it allowed a process of discernment about the value and role of CT within the established medical system whilst maintaining heterogeneity of opinion. PMID- 10812756 TI - Homoeopathy in primary care: self-reported change in health status. AB - OBJECTIVES: In recent years numerous clinical trials have been carried out to evaluate homoeopathic therapy. Another approach to evaluating homoeopathy is to investigate its impact in primary health care, that is on outpatients to a homoeopathic centre. METHODS: 609 patients on their first visit to a homoeopathic clinic from June 1995 to May 1997 received a telephone call 1 year later in which they were asked to rate their general health compared with 1 year ago. RESULTS: 1 year after first examination 73.5% of patients reported a marked or moderate improvement in their health status. Marked improvement was significantly higher in subjects who stated they were satisfied with the care received, in females, in the 0-15-year-old age group and in patients who completed therapy. CONCLUSIONS: For conditions which are not life-threatening, homoeopathy seems to meet patients' needs in primary health care. PMID- 10812757 TI - Developing a tool to measure holistic practice: a missing dimension in outcomes measurement within complementary therapies. AB - Despite the wide and continuing debate over the relevance of particular research methods to evaluate complementary therapies, less attention has been given to what outcome criteria and measures to use and/or whether these have a sufficient breadth in perspective. While complementary and alternative medicine researchers have paid heed to the importance of measuring the subjective experience of therapy users, they have left unmeasured the notion of holism, in particular, the measurement of the effect of the interaction between therapist and therapy user, and its underlying philosophy of health and healing. This paper reports on an exploratory, qualitative study aimed at developing a measure of holism and holistic practice. It draws out some key features requiring measurement and provides insight into the emerging measure. Further development work is needed before the measure can be used within research or routine practice. PMID- 10812758 TI - The BBC survey of complementary medicine use in the UK. AB - INTRODUCTION: Recent data about the use of complementary and alternative medicine (CAM) by the general population exist for Australia and the USA, but not for the UK. This study aimed at providing such data. DESIGN: Nationally representative random telephone survey. SETTING AND PARTICIPANTS: 1204 British adults were interviewed. RESULTS: 20% of the sample used CAM in the previous year. Herbalism, aromatherapy, homoeopathy, acupuncture/acupressure, massage and reflexology were the most popular treatment modalities. The main reasons for trying CAM were its perceived effectiveness, a positive inclination towards it, and its relaxing effects. On average, users spent 13.62 +/- 1.61 Pounds on CAM per month, which extrapolates to an annual expenditure of 1.6 billion Pounds for the whole nation. CONCLUSIONS: It is concluded that CAM is prevalent in the UK. Therefore, its scientific validation has become an ethical imperative. PMID- 10812759 TI - Towards better research: a discussion of some common mistakes in statistical analyses. AB - Statistical errors are common in clinical research. This paper presents a set of hypothetical but realistic scenarios in complementary medicine research. Each scenario contains at least one mistake typical of those made in analyses. These are discussed. The general principles are that charts and summary statistics can be misleading or even meaningless if not chosen carefully; objectives of the research need to be clearly defined and related to statistical hypotheses; assumptions of tests need to be checked; discussing the practical implications of observed effects is better than merely quoting P-values and data trawling should be avoided. PMID- 10812760 TI - Acupuncture, touch, and the placebo response. PMID- 10812761 TI - Primary health care transformed: complementary and orthodox medicine complementing each other. AB - Charlotte Paterson is a general practitioner and researcher at Warwick House Medical Centre in Taunton, Somerset. This is a group practice where, since 1991, complementary practitioners have worked on a private sessional basis in the centre. This collaboration was the stimulus for an ongoing research programme into various aspects of complementary medicine and primary care, and the practice is now an NHS funded Research General Practice. Charlotte Paterson has taken a keen interest in different models of integrating complementary and orthodox medicine with the aim of making complementary medical provision equitable and accessible. PMID- 10812762 TI - Commentary on Sommer et al. 'A randomized experiment of the effects of including alternative medicine in the mandatory benefit package of health insurance. AB - The study by Sommer et al. recently reported in Complementary Therapies in Medicine has been heavily criticised in Switzerland since its original publication. Its major problems are an inadequate reflection of real practice, an inadequate study design relative to the central research objective, questionable value of the applied instrument and procedure for health assessment, methodological and statistical problems, and failure to consider literature relevant to the topic. For these reasons, this experimental study does not allow an answer to its central questions as to costs and effectiveness of complementary medicine made available within Switzerland's mandatory basic health insurance provisions. We propose more practice-related, non-experimental prospective study designs to realistically answer these questions. PMID- 10812763 TI - Commentary on Sugimoto and Furnham 'The health beliefs, experiences and personality of Japanese patients seeking orthodox vs complementary medicine'. PMID- 10812764 TI - Thyrotoxic hypokalemic periodic paralysis case report and review of the literature. AB - A rare complication of hyperthyroidism consists of sudden diffuse muscle weakness associated with severe hypokalemia. The clinical presentation is similar in most respects to familial periodic paralysis; however, the therapies proven to be effective differ in the two syndromes. A case of thyrotoxic hypokalemic periodic paralysis is presented and the literature is reviewed. PMID- 10812765 TI - Use of the Nd-YAG laser improves quality of life and economic factors in the treatment of hemorrhoids. AB - PURPOSE: Hemorrhoidal disease may benefit from the use of Nd-YAG laser to decrease surgical recovery time, postoperative hospital stay and complications. METHODS: Fifty patient charts from 1993 to 1998 were reviewed retrospectively to evaluate postoperative complications and overall patient satisfaction following hemorrhoidectomy. We used the Nd-YAG laser from Surgical Laser Technologies CL60 with the ERP4 sapphire tip and the setting of 20 watts on continuous wave mode. Coagulation posthemorrhoidal excision of the remaining tissue was done using 60 watts pulse wave setting of 0.3 seconds. RESULTS: Laser treated hemorrhoidectomy patients experienced less pain than the standard hemorrhoidectomy patients. One week after surgery, the laser treated patients had 65% less pain than the standard hemorrhoidectomy patients. Painless defecation occurred earlier in the laser treated patients by five days and postoperative drainage was less than standard surgically treated patient. Surgical and hospital costs were lower by 27% and 11% respectively in the laser treated group. 88% of the laser treated patients vs 44% of the standard patients resumed work at one week after surgery. CONCLUSIONS: Nd-YAG laser treated hemorrhoid surgery patients had a quicker recovery and earlier return to work. PMID- 10812766 TI - Twin vaginal birth after cesarean. AB - OBJECTIVE: To determine both success rate and maternal-fetal outcome of vaginal birth after cesarean in twin gestations. METHODS: We identified all women from a single center attempting vaginal birth of twins after cesarean from 1988-98. Twin pairs were excluded for delivery < or = 25 weeks gestation, monoamnionicity, nonvertex twin A, or major anomaly or death of either twin. Cases were matched to the next three consecutive twin gestations attempting vaginal delivery without a prior cesarean. Variables matched were gestational age at delivery (+/- 1 week), presentations of both fetuses, labor onset (spontaneous or induced), and prior vaginal delivery (yes or no). The primary outcome was successful vaginal delivery of both fetuses. Secondary maternal outcomes included chorioamnionitis, hemorrhage requiring transfusion, hysterectomy, uterine rupture, and length of stay. Neonatal outcomes included one and five minute Apgars, NICU admission, and length of NICU stay. RESULTS: Twelve parturients were matched to 36 controls. There were no differences between the groups with respect to maternal demographics, intrapartum variables, fetal genders, birthweights, or chorionicity. Women with a prior cesarean (10/12) delivered both twins vaginally compared to 31/36 parturients without a prior cesarean (P = 1.0). There were no differences between cases and controls with respect to maternal morbidity (1/12 vs 4/36, P = 1.0), or postpartum stay (2.5 +/- 1.0 vs 2.5 +/- 2.3 days, P = .25). Neonatal outcomes were similar by birth order, except that second-born twins of cases had significantly longer NICU stays than controls (22.7 +/- 3.8 vs 10.4 +/- 7.8 days, P = .04). CONCLUSION: Twin trial of labor after cesarean is associated with a high success rate of vaginal delivery but may be associated with a more lengthy NICU stay for the second twin. PMID- 10812767 TI - Quinupristin/dalfopristin: a treatment option for vancomycin-resistant Enterococci. PMID- 10812768 TI - Primary hyperparathyroidism: successful parathyroidectomy and persistently elevated intact PTH. PMID- 10812769 TI - [Malignant schwannoma of the small intestine: a report of 2 cases]. AB - The authors report two cases of small bowel malignant schwannoma. It is an extremely rare disease and it is often difficult to diagnose malignancy. Histologic differentiation from other stromal tumors may require electron microscopy, although the preparation of immunohistochemical reactions now allows to identify forms which were previously diagnosed in different terms to be precisely classified. Diagnosis is often late and no preoperative test enables a correct clinical diagnosis to be made. The primary treatment is surgical and the resection is the only real therapy. Radiotherapy and chemotherapy are ineffective. Close postoperative observation is recommended because of the tendency to recurrence and metastasis. PMID- 10812770 TI - [A gun-shot lesion of the internal carotid. A clinical case and review of the literature]. AB - The authors report a case with penetrating zone II neck wound and internal carotid artery injury. Optimal evaluation and management with vascular injury remains controversial. A review of retrospective studies reported in the literature has demonstrated that physical examination alone may be as accurate as arteriography in detecting significant cervical vascular injuries requiring operative repair. Not unexpectedly, the results of carotid artery repair in neurologically stable patients have been excellent, in front of the results of revascularization in patients with equivocal or less-severe neurologic deficits. PMID- 10812771 TI - [The results of total thyroidectomy in Basedow's disease. The authors' personal experience of 180 cases]. AB - The therapeutic guidelines of Basedow (Graves') disease are based on three different approaches: medical treatment with anti-thyroid drugs, radio-iodine therapy and surgery. This choice has increasing consensus for the great improvement of the techniques, which at the moment are practically free of mortality risks, and have very low morbidity incidence, as well as for its positive influence on the course of the Graves' ophthalmopathy. For authors' study 180 patients affected with Graves disease, who have undergone total thyroidectomy, were retrospectively enrolled. The results show that hyperthyroidism symptoms are almost completely cured in most of the subjects, with a very low rate of complications as related to nervous (ricorrential) and parathyroid gland function. Furthermore, total thyroidectomy has resulted to have a positive impact in the clinical course of Graves' ophthalmopathy, particularly regarding some aspects of the eyes involvement, which have a dramatic effect in the "quo ad valitudinem" prognosis of these patients. PMID- 10812772 TI - [Gastric leiomyoblastomas: a clinical case report]. AB - Leiomyoblastomas are about the 10% of non epithelial gastric neoplasms. They are interesting for clinical, therapeutic and prognostic findings very different in comparison commonest epithelial tumours. The authors, after discussing aetiology, pathogenesis, clinical outline, diagnosis, histological and pathological features as well as therapeutic treatment of these neoplasms, report a case describing diagnostic and therapeutic management. PMID- 10812774 TI - [Hereditary nonpolyposis colorectal cancer (Lynch syndrome): a review of the literature and case reports]. AB - The recent observation of a new HNPCC patient case induced the Authors to review their experience with the syndrome as well as to make an up to date of the problems related to diagnosis, surgical management, surveillance and genetic counselling for such patients with a lifelong high cancer risk. Patients with HNPCC and their first-degree relatives, whose risk of early colorectal carcinoma (especially in the proximal colon) as well as a variety of extracolonic cancers (particularly endometrium, ovary, stomach, small bowel, ureter and renal pelvis) is significantly higher then that of patients with sporadic carcinoma, should be properly managed with surgery and then with endoscopic examination (ideally all life long) starting--in unaffected individuals--at early age (25 years old). Problems related to genetic counselling are considered as well. PMID- 10812773 TI - [Primary adenocarcinoma of the jejunum arising in the anastomotic loop of a Roux en-Y esophago-jejunostomy: a case report]. AB - The authors report a case of primary adenocarcinoma arising in the jejunal limb of a Roux-en-Y esophago-jejunostomy in a 76-years-old Caucasian woman. She was affected by esophageal achalasia and in another surgical department underwent gastric resection for adenocarcinoma 6 months before admission. A total gastrectomy performed was due to severity of achalasia symptoms conditioning a high degree of malnutrition. Two years after discharge the patient underwent surgery for total dysphagia, epigastric pain and inexorable weight loss. Histological response was adenocarcinoma of the long arm of Roux-en-y loop. Primary malignancies of the jejunum and ileum are extremely rare and many pathogenetic hypotheses were done but none of them received clinical or experimental demonstration. Symptoms are not specific and diagnosis is difficult and late, x-rays and endoscopy leading to it, US, CT and MNR are useful for differential diagnosis and staging. Prognosis is severe. Surgical therapy is large resection, if possible, but often it is only palliative. PMID- 10812775 TI - [The emergency management of foreign bodies: our own experience in 46 cases]. AB - Foreign bodies ingestion is a common cause of emergency endoscopic therapy. Usually ingestion is accidental for children and elderly patients, instead is voluntary in some others as psychopathic people or prisoners. The authors report their experience with 46 cases from January 1992 to December 1998: this technique is safe, with low rate of complications and hospitalization. PMID- 10812776 TI - [Reconstruction of the thoracic wall]. AB - The resection of the thoracic wall for cancer, traumas or results of radiotherapy could require a reconstruction of the same wall with prosthetic material of different nature to the purpose of protect the important intrathoracic structures, avoid the flail chest and maintain a ventilation adjusted with aesthetically acceptable results. Numerous and varied they have been and they are the materials used to such aim, but the more numerous experiences concern the reconstructions of the wall with the employment of nets of Marlex or the patch of Gore-Tex. In the complete two years eight patients have arrived at our observation in which a prosthetic reconstruction has been performed with heterologous material. In three of them has been used the net of Marlex, in five the patch of Gore-Tex of two millimeters of thickness. The prostheses have stayed well you bear and in all the patients the authors have gotten a good stabilization of the thoracic wall. PMID- 10812777 TI - [Transplantation of the intestines and bacterial translocation]. AB - Infections, sepsis and multiple organ failure syndrome are associated with high morbidity and mortality in human and experimental small bowel transplantation (SBTx). These complications are attributed to bacterial translocation demonstrated in animal and human studies. Bacterial translocation (BT) is defined as the passage of viable bacteria from the intestinal lumen to other tissues or organs. BT has been associated with different clinical and experimental situations, hemorrhagic shock, trauma, bowel obstruction, immunodepression, total parenteral nutrition, antibiotics. Although BT has been investigated in several small and large animal models of SBTx, precise information on the mechanisms involved are not available. It is possible that the operative procedure by itself may promote BT for the interaction of a number of factors such as preservation, ischemia/reperfusion, abnormal motility, lymphatic disruption and aberrant systemic venous drainage, acute or chronic rejection and antibiotic therapy. Furthermore, the potent immunosuppressive therapy used in these patients may augment the deleterious effects caused by BT. In this review we examined the existing literature concerning BT with particular regard to intestinal transplantation, to better understand the alterations in the symbiotic relationship between immunocompromised host and his gut microflora after SBTx. PMID- 10812778 TI - Can social insurance for long-term care work? The experience of Germany. AB - In 1994 Germany enacted a universal-coverage social insurance program for long term care to largely replace its means-tested system. The program has achieved many of its stated policy goals: shifting the financial burden of long-term care off the states and municipalities; expanding home and community-based services; lessening dependence on means-tested welfare; and increasing support of informal caregivers. Many of these goals were reached without exploding caseloads or uncontrolled expenditures. We examine the German long-term insurance program, focusing on issues of financing, eligibility and assessment, benefits, availability of services, and quality assurance. PMID- 10812779 TI - Long-term care insurance comes to Japan. AB - Japan has moved decisively toward "socialization of care" for the frail elderly by initiating public, mandatory long-term care insurance (LTCI) on 1 April 2000. The LTCI program covers both institutional and community-based caregiving. Everyone age forty and older pays premiums. Everyone age sixty-five and older is eligible for benefits based strictly on physical and mental disability, in six categories of need. Benefits are all services, with no cash allowance for family care, and are generous, covering 90 percent of need. Long-term costs seemed not to be a major consideration in program design. Consumers can choose the services and providers they want, including use of for-profit companies. PMID- 10812780 TI - Long-term care in the United States: an overview. AB - Although long-term care receives far less U.S. policy attention than health care does, long-term care matters to many Americans of all ages and affects spending by public programs. Problems in the current long-term care system abound, ranging from unmet needs and catastrophic burdens among the impaired population to controversies between state and federal governments about who bears responsibility for meeting them. As the population ages, the pressure to improve the system will grow, raising key policy issues that include the balance between institutional and noninstitutional care, assurance of high-quality care, the integration of acute and long-term care, and financing mechanisms to provide affordable protection. PMID- 10812781 TI - Rearranging the compartments: the financing and delivery of care for Australia's elderly. AB - Aged care policy in Australia underwent rapid change following the 1996 change of federal government, although continuing to emphasize changing the balance of aged care away from residential care toward community-based care and improving quality of care. This paper examines these policy directions with reference to two specific areas: the differentiation of funding arrangements for the care and accommodation components of residential care, and the targeting of services in community care. In both cases, funding arrangements have been used as a prime mechanism for redrawing the boundaries between different components and levels of care. This process of compartmentalization appears likely to increase the diversity of the Australian aged care system in the future. PMID- 10812782 TI - New Zealand: long-term care in a decade of change. AB - Long-term care in New Zealand incorporates a mix of public and private funding and provision. After a decade of structural change, the purchasing of almost all publicly funded health and social care is now the responsibility of one central agency. Services for older persons are poorly integrated, and there are problems of access to and quality of some services. Efforts are being made to address these problems. The challenge now is to ensure that this groundwork is not lost amid the turmoil of yet another round of restructuring by an enthusiastic, newly elected government. PMID- 10812783 TI - A tale of two systems: the changing academic health center. AB - Major changes in academic health centers (AHCs) may not be confined to the United States. Both Partners HealthCare System in Boston and University College London School of Medicine/University College Hospital Trust in London have recently undergone mergers, downsizing, and cost cutting on unprecedented scales. A comparison of the recent histories of these eminent AHCs reveals striking similarities in the clinical and academic pressures bearing down upon them and in their responses. It also reveals important differences in their situations and actions, traceable in large part to the contrasting roles of governments and markets in the health care economies of these two countries. PMID- 10812784 TI - In pursuit of an improving National Health Service. AB - The British National Health Service (NHS) before its 1990s internal-market reforms was a gridlock of perverse incentives. The internal market, an attempt to introduce some market incentives, stimulated much innovation in primary care commissioning and practice improvement and led to increased efficiency. However, its effects were quite limited, because the essential conditions for a market to operate were not fulfilled. There now exists a crisis of confidence in the quality of care in the NHS. It is doubtful whether a culture of innovation, efficiency, and good customer service is possible in a public-sector monopoly whose services are in excess demand and whose units do not get more resources for caring for more patients. It also is doubtful whether the NHS can modernize without consumer choice, competition, and more resources. PMID- 10812785 TI - The modern NHS: an underfunded model of efficiency and integration. PMID- 10812786 TI - Judging the success of reform: the view from within. PMID- 10812787 TI - Australia's balance between public and private arrangements. PMID- 10812788 TI - The politics of health system reform. PMID- 10812789 TI - The future of the NHS: confronting the big questions. PMID- 10812790 TI - Questioning the role of the private market in the British NHS. PMID- 10812791 TI - Restoring the status of an icon: a talk with Canada's Minister of Health. Interview by John K. Iglehart. PMID- 10812792 TI - Caring for the frail elderly: an international review. PMID- 10812793 TI - Health spending and outcomes: trends in OECD countries, 1960-1998. PMID- 10812794 TI - Health plan switching: choice or circumstance? PMID- 10812795 TI - What would happen if large firms offered MSAs? AB - This paper reports the results of a survey of more than 500 health benefit specialists about the advice they would give to medium-size and large employers on offering a tax-advantaged medical savings account (MSA). About 42 percent of respondents would recommend an MSA combined with a catastrophic health plan, while a third would advise against such a plan. When presented with a specific example of an MSA package that would be attractive to a large fraction of workers, the percentage of benefit specialists favoring adding an MSA option rose to 74 percent. However, respondents generally did not believe that most workers would choose the MSA, especially if the alternative were a health maintenance organization (HMO). PMID- 10812796 TI - Where does performance measurement go from here? PMID- 10812797 TI - Leveling the playing field: managed care enrollment and hospital use, 1987-1996. PMID- 10812798 TI - My twelve years in the U.K. health system. PMID- 10812799 TI - Population aging: a comparison among industrialized countries. AB - Increasing longevity and declining fertility rates are shifting the age distribution of populations in industrialized countries toward older age groups. Some countries will experience this demographic shift before others will. In this DataWatch we compare the effects of population aging on health spending, retirement policies, use of long-term care services, workforce composition, and income across eight countries: Australia, Canada, France, Germany, Japan, New Zealand, the United Kingdom, and the United States. International comparisons suggest that the United States is generally well positioned to cope with population aging; however, three areas should be carefully monitored: heavy reliance on private-sector funding of retirement, coverage of pharmaceuticals for the elderly, and a high proportion of private long-term care financing. PMID- 10812800 TI - Population aging in developing countries. AB - Issues related to population aging--Social Security and pension reform, health care financing and provision, and long-term care--have long been the subject of public debate in the industrialized countries of Europe and North America. Economically less developed regions have been slower to adopt aging as a major public policy concern, despite the fact that older populations in many developing countries are growing more rapidly than are those of industrialized nations. Awareness of issues concerning older populations remains low in many nations, even as the absolute numbers of elderly persons double and even triple. This DataWatch considers trends in the basic demography of aging and mortality/health considerations for developing countries. PMID- 10812801 TI - Coping with aging: international challenges. AB - This DataWatch discusses trends in functional health status among the elderly in several countries. Life expectancy at an advanced age has increased, and there is some evidence of lower prevalence of severe disability and a trend toward deinstitutionalization. These changes may result in fewer severely disabled persons, but the impact is less clear from the perspective of public finances, because of the increased need for formal home care and the relative subsidization of institutions. The implications for health care need further research but also may point to increased expenditures. PMID- 10812802 TI - The elderly in five nations: the importance of universal coverage. AB - This paper reports 1999 survey results on the population age sixty-five and older in five nations--Australia, Canada, New Zealand, the United Kingdom, and the United States. The majority of respondents were generally satisfied with the quality, affordability, and availability of health services in their nations. In many measures of access to and cost of care, the United States looks much like the other nations surveyed. However, as the elderly view their health systems, the direction they have taken in recent years with respect to caring for the elderly, and the future affordability of care in old age, U.S. respondents tended to be more pessimistic than were those in other nations. PMID- 10812803 TI - Using 'linkage and exchange' to move research into policy at a Canadian foundation. PMID- 10812804 TI - Strategic philanthropy: principles put into practice. PMID- 10812805 TI - The United States confronts the policy dilemmas of an aging society. PMID- 10812806 TI - Outpatient pharmaceuticals and the elderly: policies in seven nations. PMID- 10812807 TI - Tax credits for insurance: a viable option. PMID- 10812808 TI - Insurance tax subsidies: you get what you pay for. PMID- 10812809 TI - A market for control of nonprofits. PMID- 10812811 TI - Refuting the 'accidental system'. PMID- 10812810 TI - Correcting the terminology in study of preventive care. PMID- 10812812 TI - Gastroenterology on the Internet--II. PMID- 10812813 TI - Current blood transfusion practices in India. PMID- 10812814 TI - Influence of intragastric perfusion of aqueous spice extracts on acid secretion in anesthetized albino rats. AB - BACKGROUND: The effect of spices on gastric acid secretion is variable. Their mechanism of action is also not well established. AIM: To study the effect of spices on gastric acid secretion in anesthetized rats. METHODS: Aqueous extracts (10% w/v) of red pepper (Capsicum annuum), fennel (Foeniculum vulgare), omum/ajwan (Carum copticum), cardamom (Elettaria cardamomum), black pepper (Piper nigrum), cumin (Cuminum cyminum) and coriander (Coriandrum sativum) were prepared. The stomach of pentobarbitone-anesthetized rats was perfused at 0.15 mL/min with aqueous extracts of spice or acetylcholine (1 microgram/mL or 10 micrograms/mL solutions, in 40 min blocks, twice in each experiment bracketed by saline perfusions. The acid content in the samples was estimated by titration with 0.1N NaOH with phenolphthalein as indicator. Atropine 1 microgram/mL was added to the perfusion fluid in 28 experiments. In 32, acute gastric mucosal injury was induced by leaving aspirin 125 mg/Kg in the stomach for 2 h before perfusion. RESULTS: All the spices tested increased acid secretion in the following declining order: red pepper, fennel, omum, cardamom, black pepper, cumin, coriander. Red pepper increased acid secretion (mean [SEM] 0.93 [0.16] mL 0.1N HCl) to about 7 times the basal secretion (0.14 [0.05]; p < 0.005). The increase in acid secretion by the other spices was as follows: fennel 0.42 (0.11) mL 0.1 N HCl from basal secretion (0.12 [0.03]) (p < 0.02); omum 0.33 (0.05) from 0.09 (0.02) (p < 0.01); cardamom 0.28 (0.04) from 0.10 (0.03) (p < 0.005); black pepper 0.19 (0.03) from 0.04 (0.01) (p < 0.005); cumin 0.12 (0.02) from 0.08 (0.01) (p < 0.05); coriander 0.18 (0.03) from 0.09 (0.02) (p < 0.005). Atropine abolished the acid secretion induced by acetylcholine and significantly reduced acid induction by red pepper, omum and coriander, but not that by fennel. In experiments with aspirin-induced mucosal injury the basal acid secretion was low; acid secretion by red pepper and fennel was reduced significantly, but not that by acetylcholine. Cumin and coriander increased acid secretion in injured stomachs. CONCLUSION: The spices tested increased gastric acid secretion, in some by a cholinergic mechanism but by other mechanism(s) as well. Red pepper produced maximum increase in acid secretion, but this was significantly reduced in injured stomachs. Cumin and coriander increased gastric secretion in injured stomachs. PMID- 10812815 TI - Proliferating cell nuclear antigen and nucleolar organizer region for differential diagnosis of dysplasia and adenocarcinoma in gastric biopsies. AB - INTRODUCTION: The value of proliferating cell nuclear antigen (PCNA) and nucleolar organizer region (AgNOR) for differential diagnosis of normal mucosa, dysplasia and adenocarcinoma in gastric endoscopic biopsies, and correlation between these two methods were evaluated. METHODS: 15 gastric endoscopic biopsy samples from normal mucosa, 15 from areas of dysplasia and 15 from low grade adenocarcinoma were studied. AgNOR and PCNA immunostaining were applied to paraffin sections. RESULTS: Mean AgNOR value and PCNA-labeling index were the lowest in normal mucosa and the highest in adenocarcinoma. Mean (SD) AgNOR numbers were 2.9 (0.3) in normal mucosa, 5.9 (1.7) in dysplasia and 15.7 (2.8) in adenocarcinoma. PCNA-labeling index was 2.4 (1.1) in normal mucosa, 27.5 (4.6) in dysplasia and 42.1 (5.3) in adenocarcinoma. The differences between normal mucosa and dysplasia, and between dysplasia and adenocarcinoma were significant (p < 0.001). Overlapping values were observed in AgNOR counts between normal mucosa and dysplasia, and in PCNA-labeling indices between dysplasia and adenocarcinoma. No correlation was found between AgNOR and PCNA. CONCLUSION: Though mean AgNOR values and PCNA indices were significantly different between normal mucosa, dysplasia and adenocarcinoma, these could not be used in differential diagnosis because of overlapping values between groups. PMID- 10812816 TI - Evaluation of endoscopy-based diagnostic methods for the detection of Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori is an etiological factor in duodenal ulcer. Few studies have objectively assessed the accuracy of diagnostic methods for the detection of H. pylori. METHODS: The sensitivity and specificity of histology, touch smear, rapid urease test (RUT) and brush cytology of endoscopic antral biopsy from patients with duodenal ulcer were compared. Forty-nine patients were evaluated before, and 34 after, eradication therapy. Each time, sampling was done for all 4 tests. The infection status for each sample was established by a positive concordance of results of three of four tests. RESULTS: The highest degree of agreement was between RUT and cytology (kappa = 0.69). Brush cytology (100%) followed by RUT (94.5%) were the most sensitive tests. Histology had the highest specificity (89.3%). A combination of RUT or brush cytology with histology had the maximum chance of detecting H. pylori. As single tests, brush cytology and touch smear had high diagnostic accuracies with a Youdin J value of 1.79 and 1.78, respectively. CONCLUSION: The best method for diagnosis of H. pylori is a combination of the rapid urease test or brush cytology with histology. Brush cytology or touch smear are diagnostic tests of choice if a single test is desired. PMID- 10812817 TI - Blood transfusion practices in India: results of a national survey. AB - BACKGROUND: Blood transfusion may lead to serious clinical consequences for the recipient, if the transfused blood is not safe. To assess the functioning of the blood banks in India, a nation-wide, questionnaire-based study was conducted between November 1995 and November 1996 under the auspices of the Indian Association for Study of the Liver. METHODS: Of 604 blood banks in 31 states and union territories to whom the questionnaires were sent, responses wereobtained from 78 (13%) blood banks in 17 (54.8%) states, providing information on 275,000 donors. RESULTS: A majority (58%) of donors in these blood banks were replacement donors, followed by voluntary (39.3%) donors. About 87% of the respondent blood banks screen blood for hepatitis B, 95% for HIV, 94% for syphilis, 67% for malaria, and only 6% for hepatitis C. Marked heterogeneity in the test methods was observed with only 13% using ELISA kits for HBsAg. Only 21% of the blood banks prepare blood-derived components. Feedback to the blood banks on the occurrence of transfusion-associated hepatitis is given on less than 40% of occasions. CONCLUSIONS: Testing for transfusion-transmitted infections is unsatisfactory and poorly regulated in India. Reporting of adverse events after transfusion is poor and no stringent donor deferral system exists. PMID- 10812818 TI - Dyspepsia in the general population in Jordan. AB - OBJECTIVE: To assess the prevalence of dyspeptic symptoms in the general population of Jordan. METHODS: We selected families of final year students at the two medical schools in Jordan to represent the general population. The students were instructed to complete questionnaires for all their family members and all members of the close family above the age of 15 years living in the area. The questionnaire enquired about dyspeptic symptoms in the previous 12 months. Information about any chronic disease, medications used frequently, previous operations and cigarette smoking was obtained. RESULTS: Of 2254 completed questionnaires, dyspeptic symptoms were reported in 1354 (60.1%) during the previous 12 months. Heartburn was present in 1111 (49.3%), abdominal distension in 1224 (54.3%), recurrent abdominal pain with ulcer-like characters in 450 (20%), discomfort following meals in 316 (14%), symptoms related to fatty foods in 359 (15.9%) and symptoms related to ingestion of lactose-containing foods in 96 (4.3%). CONCLUSION: Dyspeptic symptoms are highly prevalent in Jordan. This may have an impact on the economy and health resources. PMID- 10812819 TI - Safety and immunogenicity of indigenous recombinant hepatitis B vaccine (Shanvac B) in comparison with commercially available vaccine. AB - AIM: To assess the clinical safety, reactogenicity and immunogenicity of an indigenously developed recombinant hepatitis B vaccine (Shanvac-B; Shantha Biotechnics) and to compare it with another commercially available vaccine (Engerix-B, SmithKline Beecham) in healthy adults. METHODS: 120 healthy adults randomLy received 20 micrograms of either Engerix-B (Group A; n = 61) or Shanvac B (Group B; n = 59) in 0, 1, 2 months schedule. Anti HBs was assessed using commercially available AUSAB kits (Abbott Laboratories) one month after each dose. RESULTS: Protective seroconversion rates after first, second and third dose were 10%, 62.7% and 91.4%, respectively in Group A and 22.4%, 68.9% and 96.4% in Group B, respectively. The geometric mean titer (GMT) after the third dose was significantly high in Group B (419 mIU/mL) than in Group A (140 mIU/mL; p < 0.001). The GMT was significantly higher in women in both the groups. The indigenous vaccine was found to be clinically safe and well tolerated without significant side effects. CONCLUSION: The recombinant hepatitis B vaccine (Shanvac-B) developed in India is safe, well tolerated, and highly immunogenic, with high seroconversion and GMT response. PMID- 10812820 TI - Epidemiology of digestive tract cancers in India. VI. Projected burden in the new millennium and the need for primary prevention. AB - The incidence rates of most digestive cancers in India are moderate or low. The highest rates are recorded in the urban population of Mumbai and the lowest in the rural population of Barshi in Maharashtra state. The rates will rise as the life expectancy of Indians increases along with urbanization and, within the next few decades, may reach those recorded in Indians living abroad. Based on available population data, we estimate that in the year 2001 there will be approximately 145,000 new cases of digestive cancers in India. In men, the esophagus would be the commonest site (n = 24,925), followed by the stomach (23,100), rectum (10,462), liver (8812), colon (8004), pancreas (5757) and gall bladder (3967). In women, the esophagus would be the commonest site (n = 18,608), followed by the stomach (11,890), gall bladder (7360), rectum (6983), colon (6115), liver (4227) and pancreas (3435). The incidence of cancers of the esophagus and stomach is declining spontaneously in India. It may be possible to accelerate this by reducing the use of tobacco and improving the diet. At the same time the incidence of cancers of the colon, pancreas, liver and gall bladder is rising, largely due to urbanization that leads to major changes in the diet and personal habits. A preventive approach is needed by public health education. Indians should be encouraged to retain their traditional protective diets, eat more fruits and vegetables, do more physical activity, and abstain from tobacco. Gastroenterologists can also help in secondary prevention by screening high-risk individuals, e.g., patients with chronic liver disease for liver cancer and relatives of patients with familial bowel cancer. PMID- 10812821 TI - Jejunal stricture complicating acute mesenteric venous thrombosis secondary to protein C deficiency and factor V Leiden gene mutation. AB - We report a patient with high small intestinal obstruction due to ischemic stricture, secondary to acute mesenteric venous thrombosis. Tests done for hypercoagulability revealed dual etiology (protein C deficiency and factor V Leiden gene mutation). The patient underwent surgical resection and has been on anticoagulation treatment. At 2 years' follow up, she remains asymptomatic. PMID- 10812822 TI - Laparoscopic management of hydatid cyst of liver. AB - We describe the laparoscopic excision of a hydatid cyst in the liver. During the procedure, done after treatment with the scolicidal agents praziquantel and albendazole, care was taken to prevent spillage of scolices during evacuation of contents and to excise the entire germinal epithelium. The patient had no immediate or short-term complications and is asymptomatic 3 months later. PMID- 10812823 TI - Intestinal pseudo-obstruction due to neurofibromatosis responding to cisapride. AB - Gastrointestinal involvement in neurofibromatosis presenting with mechanical obstruction, hemorrhage or intussusception is known. Small bowel dysmotility and intestinal pseudo-obstruction due to neurofibromatosis is rare. A 23-year-old man with classical neurofibromatosis presented with intermittent episodes of intestinal pseudo-obstruction, small bowel bacterial overgrowth and steatorrhea. The patient had good symptomatic improvement with cisapride. PMID- 10812824 TI - Dapsone hypersensitivity syndrome with coexisting acute hepatitis E. AB - A 14-year-old girl presented with fever, generalized lymphadenopathy, skin rash and hepatitis after starting dapsone. All abnormalities reversed with institution of prednisolone therapy after discontinuation of dapsone. The hepatic involvement was of hepatocellular type; it was associated with IgM anti-HEV antibodies, suggesting coexisting acute hepatitis E. We believe a causal link between the hepatotrophic viruses and dapsone hypersensitivity syndrome could exist. PMID- 10812825 TI - Palliation of malignant colonic obstruction by self-expandable metal stent. AB - Advanced obstructive colorectal cancer is routinely treated by surgical colostomy. Self-expandable metal stents are a promising alternative. We report the use of an expandable metal stent to relieve colonic obstruction in an elderly lady with advanced colorectal malignancy. PMID- 10812826 TI - Isolated massive splenic peliosis. AB - We report a patient with isolated splenic peliosis. She presented with massive splenomegaly; contrast-enhanced CT scan showed multiple hypodense, well circumscribed shadows in the spleen. Splenectomy specimen showed multiple cysts with white fibrous walls ranging from 1-4 cm in size, containing gelatinous fluid. The patient is asymptomatic at 6 months' follow up. PMID- 10812827 TI - Ulcerative colitis with sagittal sinus thrombosis with normal coagulation profile. AB - Thromboembolic events are serious complications in patients with inflammatory bowel diseases. We describe a 30-year-old man with ulcerative colitis complicated by sagittal sinus thrombosis with normal coagulation profile; he achieved clinical remission with subcutaneous heparin. PMID- 10812828 TI - Retroperitoneal teratoma presenting as acute abdomen in an elderly person. AB - A 56-year-old man presented with acute abdomen. Clinically, he was diagnosed as having perigastric abscess. On exploration, a retroperitoneal cystic teratoma was encountered. Postoperatively, he recovered uneventfully and has no residual disease two years later. PMID- 10812829 TI - Hepatocellular carcinoma in association with membranous obstruction of inferior vena cava. AB - We report a patient with hepatocellular carcinoma (HCC) with membranous obstruction of the inferior vena cava (IVC). He underwent balloon dilatation of the IVC with good results. The HCC was managed by chemoembolization followed by resection. At follow up of eleven months the patient is asymptomatic. PMID- 10812830 TI - Hemoperitoneum following rupture of ectopic varix along splenorenal ligament in extrahepatic portal vein obstruction. AB - A 29-year-old man with extrahepatic portal vein obstruction who underwent variceal eradication by sclerotherapy six years ago, was admitted with hypotension and abdominal pain. Abdominal paracentesis yielded frank blood. Laparotomy showed bleeding from a large ectopic vessel along the splenorenal ligament. The vessel was ligated and the patient recovered. PMID- 10812831 TI - Intestinal intussusception in children: a review of 70 cases. PMID- 10812832 TI - Suboptimal response to commercially available kits for Helicobacter pylori eradication. PMID- 10812834 TI - Work in OECD on chemical safety: approaches for human risk assessment. PMID- 10812833 TI - Reprocessing of endoscopes requires guardians. PMID- 10812835 TI - Impairment of neurobehavioral function and color vision loss among workers exposed to low concentration of styrene--a review of literatures. AB - Recently many studies on the health effects of workers who exposed to low concentration styrene have been published. In this paper firstly we mentioned the reasons why nervous system was critical organ for evaluating the toxicity of organic solvents both in the acute and chronic exposure phases. Then we indicated how neurobehavioral test batteries were useful to detect subclinical adverse health effects of workers exposed to organic solvents. Secondly we reviewed many epidemiological studies on the impairment of neurobehavioral function, i.e., perceptual speed, memory, cognition, personality and mood, and the loss of color vision among styrene workers. Finally, we showed our recent data on the relationship between color vision loss and the concentration of urine metabolites among styrene workers. It can conclude that styrene may cause damage on nervous system, even at the low exposure level such as 50 ppm which used to be the Threshold Limit Value (TLV) recommended by Japan Society for Occupational Health (until 1999) and American Conference of Industrial Hygienists (until 1997). PMID- 10812836 TI - Biomarkers of lead exposure. AB - Biomarkers of exposure, effect, and susceptibility are reviewed in relation to lead exposure. Of the biomarkers of lead exposure, blood lead (Pb-B), mainly red cell lead, is a representative of soft tissue lead, and most widely used as measures of body burden and absorbed (internal) doses of lead. Urine lead (Pb-U) as well as plasma lead (Pb-P) increases exponentially with increasing Pb-B under a steady-state situation and is a reflection of recent exposure. The amount of lead in plasma and urine (MPb-P and MPb-U) after administration of a chelating agent (e.g. CaEDTA) can be useful for biomarkers of internal exposure of lead, reflecting the mobilizable pool of lead which consists of mainly blood and soft tissue lead with only a small fraction derived from bones. The critical effects in bone marrow arise mainly from the interaction of lead with some enzymatic process responsible for heme synthesis. The effects can be used for the biomarkers of effects. They are the inhibition of delta-aminolevulinic acid dehydratase (ALAD) and the variation in some metabolite concentrations (e.g. delta-aminolevulinic acid in urine (ALA-U), blood (ALA-B) or plasma (ALA-P), coproporphyrin in urine (CP), zinc protoporphyrin (ZP) in blood). The activities of pyrimidine nucleotidase (P5'N) and nicotinamide adenine dinucleotide synthetase (NADS) in blood are also decreased in lead exposure, and nucleotide contents in blood is altered in lead exposure. These effects of lead on human can be also useful biomarkers of effect. The differences in levels of heme precursors between two types of ALAD genotypes might be attributable to those in the affinity of different ALAD isozymes to lead. ALAD1 homozygotes have higher levels of ZP and ALA in comparison with ALAD2 carriers at the high lead exposure, suggesting that ALAD1 homozygotes might be more susceptible for disturbance in heme biosynthesis by lead than ALAD2 carriers. PMID- 10812837 TI - Polymorphism of drug-metabolizing enzymes in relation to individual susceptibility to industrial chemicals. AB - Polymorphism and the induction/inhibition of drug-metabolizing enzymes, such as cytochrome P450, aldehyde dehydrogenase (ALDH), glutathione S-transferase (GST), N-acetyltransferase (NAT), and NAD(P)H-quinone oxidoreductase (NQO1), were reviewed in relation to susceptibility to disease and to inter-individual difference in biological monitorings. A number of genetic and acquired factors can influence the susceptibility of an individual to chemicals, creating a so called predisposition. Most cases in which genetic factors were present resulted from polymorphism of drug-metabolizing enzymes. However, conflicting reports have appeared on the relationship between polymorphism and risk of disease; in some cases, biologically plausible mechanisms linking genotypes and disease are not yet in evidence. Current findings based on biological monitoring of chemicals are insufficient to evaluate the relationship between genetic polymorphism and acquired risk when exposure has occurred in an occupational area. Investigation of such situations has generated data implicating GSTT1, GSTM1, NAT2, and NQO1 polymorphisms in biological monitoring of some chemicals; the ALDH2 polymorphism is the likely link between the genotype and the metabolism of low molecular aliphatic aldehydes. Although this polymorphism is limited in the case of Japanese as well as other Asian subjects, the inhibitors of ALDH2 activity such as trichloroethylene may produce a false polymorphism of this gene. As to the effect of factors influencing acquired predisposition, such as ethanol intake, intake of low carbohydrate diet or diabetes, corroborative epidemiological studies may be further required. PMID- 10812838 TI - Occupational and environmental toxicology of mercury and its compounds. AB - Mercury exists in various chemical forms. The important forms from a toxicological viewpoint are the metallic form, also called the elemental form, the divalent inorganic forms and methylmercury compounds. Elemental (Hg0) mercury has a high vapor pressure and the vapor causes a number of cases of poisoning via inhalation. Classical mercury poisoning is characterized by a triad of signs, namely tremors, erethism and gingivitis. Mercurial erethism, which is characterized by behavioral and personality changes such as extreme shyness, excitability, loss of memory, and insomnia are also observed. Recently, the effects of mercury exposure at levels around 0.05 mg/m3 or lower have been of concern and may include minor renal tubular damage, increased complaints of tiredness, memory disturbance and other symptoms, subclinical finger tremor, abnormal EEG by computerized analysis and impaired performance in neurobehavioral or neuropsychological tests. Abnormal gait, dysarthria, ataxia, deafness and constriction of the visual field are typical of the symptoms of methylmercury poisoning observed in Minamata and Iraqi outbreaks, as well as in occupational methylmercury poisoning cases. Furthermore, an infant born to a mother with excessive methylmercury consumption showed various neurological disturbances and delayed development. Since several populations are believed to be still exposed to methylmercury through the consumption of fish and sea mammals, neurobehavioral deviations in children of these populations have recently been investigated. PMID- 10812839 TI - Carcinogenicity and other health effects of acrylonitrile with reference to occupational exposure limit. AB - The occupational exposure limit for acrylonitrile (AN) has been set by many organizations on the basis of its carcinogenicity. However, recent epidemiological studies do not afford evidence supporting the hypothesis that AN is carcinogenic to humans. Review of the 18 published cohort studies revealed that, although there is not adequate evidence in humans for carcinogenicity of AN, the possibility of a causal association between high exposure to AN and lung cancer in humans cannot be excluded. It was pointed out that carcinogenic potential of AN may be weak, if any, to humans, and the current occupational exposure limit (OEL) for AN of 2 ppm was evaluated as appropriate in view of AN exposure levels reported by epidemiological studies. Based also on review of the literature on health effects other than carcinogenicity, it was concluded that the current OEL for AN is a reasonable value and there is no need for a revision at present. PMID- 10812840 TI - Blood lead level to induce significant increase in urinary delta-aminolevulinic acid level among lead-exposed workers: a statistical approach. AB - The present study was initiated to examine the quantitative relationship between blood lead (Pb-B) and urinary delta-aminolevulinic acid (ALA-U) among Pb-exposed workers, and to find a threshold Pb-B level to induce an increase in ALA-U. For this purpose, pairs of venous blood and spot urine samples were collected from 8,274 men and 5,856 women (14,130 workers in total) who were occupationally exposed to inorganic lead. The blood and urine samples were analyzed for Pb-B and ALA-U by atomic absorption spectrometry and colorimetry, respectively, and the correlation between pairs of measures were subjected to statistical analysis. The assumption of the 3rd degree regression for correlation gave a substantially greater correlation coefficient (0.645 for men and 0.619 for women) than 1st or 2nd degree regression, whereas only very small improvement in the coefficient was achieved with 4th to 6th degree ones. Logarithmic conversion of the parameters was not effective in improving the correlation. The assumption of the 3rd degree regression followed by calculation of the local minimum gave 22, 29 and 23 micrograms/100 ml Pb-B for men, women, and men + women, respectively, as the threshold Pb-B to induce ALA-U increase. Pb-B to elevate ALA-U to the 95% upper normal limit (8 mg/l, common to men and women) was 62, 50 and 58 micrograms/100 ml for men, women and men + women, respectively. The validity of the 3rd degree regression assumption as a tool to calculate a threshold from experimental or epidemiological data is discussed. PMID- 10812841 TI - Decrease of glucose utilization rate in the spinal cord of experimental 2,5 hexanedione poisoning rats: application of microdetermination technique. AB - To examine the effects of 2,5-hexanedione (HD) on the glucose metabolism in the spinal cord, glucose utilization rate (GUR) and distribution volume of glucose (DV) were measured in the white matter and anterior horn of the spinal cord in 8 rats exposed to HD for 4 weeks and in 10 control rats. The GUR and DV were determined by the quantitative microdetermination method using non-tracer 2 deoxyglucose based on the three-compartments model of Sokoloff. GURs in the white matter and anterior horn and DV in the anterior horn in the HD-exposed rats were significantly lower than those in the control rats. In the multiple regression analysis, GUR in the white matter of HD-exposed rats was significantly related to blood HD concentration. It is suggested that the decrease of GUR in the white matter is a major effect of HD in the spinal cord. PMID- 10812842 TI - Urinary phenylmercapturic acid as a marker of occupational exposure to benzene. AB - A hand-saving HPLC method to measure urinary phenylmercapturic acid (PMA) was developed which allows about 35 PMA determinations per day. The method involves conversion of pre-PMA to PMA by the addition of sulfuric acid to a urine sample, extraction into an ether-methanol mixture followed by condensation under a nitrogen stream. The condensate was introduced to a ODS-3 column in a HPLC system, and PMA in the column was eluted into a mobile phase of acetonitrile: methanol: perchloric acid: water. The elution of PMA was monitored at 205 nm. One determination will be completed in 40 min. The method was applied to analysis of end-of-shift urine samples from 152 workers exposed up to 210 ppm benzene, 66 workers exposed to a mixture of benzene (up to 116 ppm) and toluene + xylenes (up to 118 ppm), and 131 non-exposed controls of both sexes. A linear regression was established between time-weighted average intensity of exposure to benzene and urinary PMA. From the regression, it was calculated that urinary PMA level will be about 6.4 mg/l after 8-hour exposure to benzene at 100 ppm, and that PMA in urine accounted for about 0.1% of benzene absorbed. No effects of sex, age, and smoking habit of individuals were detected, and the effect of co-exposure to toluene + xylenes at the levels comparable to that of benzene was essentially nil, which indicates an advantage of PMA as a benzene exposure marker over monoto tri-phenolic metabolites or t,t-muconic acid. PMID- 10812843 TI - Circadian rhythms of seven heavy metals in plasma, erythrocytes and urine in men: observation in metal workers. AB - To elucidate circadian rhythms (variation within a day) of 7 toxic or essential metals in plasma and erythrocytes in relation to the rhythms in urine in men, 19 male metal foundry workers were examined; they were exposed to lead (Pb), zinc (Zn) and copper (Cu) occupationally but separated from the exposure during the study. Circadian rhythms were found for plasma concentration of Pb, cadmium (Cd), Zn, Cu and chromium (Cr) in the workers. Circadian rhythms were also found for Pb, inorganic mercury (Hg), Zn and Cr in erythrocytes and for all metals except Zn in urine. Both the plasma and urinary levels of Pb, Cd, Cu and Cr together with urinary excretion of Mn and creatinine tended to decrease during night hours; both the erythrocyte and urinary levels of Hg together with both the plasma and erythrocyte levels of Zn tended to increase during morning hours. The results of profile analysis suggested that the circadian rhythm of Pb in urine was affected more strongly by its plasma and erythrocyte rhythms than by the rhythm of creatinine in urine, i.e. the rhythm of glomerular filtration; the urinary rhythms of Cd, Cr, and Mn were affected more by the creatinine rhythm; and the urinary rhythm of Cu was affected by both its plasma and creatinine rhythms. On the other hand, the urinary rhythm of Hg was assumed to be independent of the creatinine rhythm and be affected by its erythrocyte rhythm. The present study suggested that different biological limit values might be needed for different hours of the day especially for shift workers who are exposed to various heavy metals. Instead, further studies should be conducted to find the adjustment methods by which no circadian rhythms are discerned. PMID- 10812844 TI - Examination of lacZ mutant induction in the liver and testis of Muta Mouse following injection of halogenated aliphatic hydrocarbons classified as human carcinogens. AB - Possible induction of lacZ mutation was examined in the liver and testis of Muta Mouse following the administration of carcinogenic halogenated compounds, namely 1,2-dichloroethane (DCE), 1,2-dibromoethane (DBE), carbon tetrachloride, or 1,2 dibromo-3-chloropropane (DBCP). Slight increases were observed on the mutant frequency in the testis DNA isolated from the mice 14 days after treatment with DBCP at 40 mg/kg or with DBE at 60 mg/kg but not in the liver. Further investigation was necessary to confirm the mutation induction by these chemicals in the testis including experiments with longer sampling intervals. No increase was detected in the frequency following DCE administration of single doses of up to 150 mg/kg or of consecutive injections of up to 280 mg/kg. Marginal but biologically insignificant responses were observed in the liver from the carbon tetrachloride exposed mice. The present results suggest that these carcinogenic chemicals are less efficient for induction of gene mutation in the liver of Muta Mouse. PMID- 10812845 TI - Alteration of glucose metabolism in the spinal cord of experimental lead poisoning rats: microdetermination of glucose utilization rate and distribution volume. AB - To examine the effects of lead on glucose metabolism in the spinal cord, glucose utilization rate (GUR) and distribution volume of glucose (DV) in the anterior horn and white matter were determined in 9 rats exposed to lead for 4 weeks and 10 control rats. The GUR and DV were determined by the quantitative microdetermination method using non-tracer amount of 2-deoxyglucose based on the three-compartments model of Sokoloff. The GUR and DV in the anterior horn in the lead-exposed rats were significantly lower than those in the controls. It is thus suggested that glucose metabolism, as measured by the GUR and DV, in the anterior horn is inhibited by lead; the anterior horn cells seem sensitive to lead neurotoxicity. PMID- 10812846 TI - Inhibitory effects of heavy metals on transcription factor Sp1. AB - Heavy metals are expected to affect the biological activity of various metal containing proteins, including transcriptional regulators. We studied the effects of several heavy metal ions on the DNA-binding activity of a Zn-finger transcription factor, Sp1. With respect to both DNA elements through which Sp1 acts positively and negatively, Cd2+ inhibited DNA-binding of Sp1 at 20 microM or higher, while the toxic effect of Zn2+ was obvious only at more than 200 microM. Inhibition was also apparent for Cu2+ but less remarkable for Hg2+. The inhibition by Cd2+ was relieved by the addition of Zn2+ at much lower concentrations than that of Cd2+. These results suggest that the toxic potential of heavy metals could be largely influenced by the intracellular Zn2+ concentration. PMID- 10812847 TI - Estimation of the lethal toluene concentration from the accidental death of painting workers. AB - To determine the potentially lethal level of thinner concentration in the air, we measured the concentration of toluene in the blood and others of three patients who suffered severe acute thinner intoxication between 4 January 1996 and 21 April 1997 in Ube city. The concentration of toluene in blood were 30.2 mg/L in died patient, and 13.7 mg/L and 17.5 mg/L in recovered patients, respectively. By extrapolation from the results of our previous toxicokinetic research on toluene poisoning in anesthetized dogs, the fatal concentration of toluene was estimated to be approximately 1800 to 2000 ppm for 1-hour exposure. PMID- 10812848 TI - Relation of body weight change to changes in atherogenic traits; a study of middle-aged Japanese obese male office workers. AB - We examined the effect of body weight change on the modification of atherogenic risk factors in 296 middle-aged obese male office workers without medication for hypertension, dyslipidemia, hyperuricemia or diabetes mellitus. During a 1-year education program, 39.2% of the participants could reduce their weight, and the percentage of those who lost 2 kg or more was only 17.7%. Concomitant with the decrease of weight, however, the levels of systolic and diastolic blood pressures, total cholesterol, triglyceride, uric acid and hemoglobin A1c and the ratio of total cholesterol to high-density lipoprotein (HDL) cholesterol decreased significantly, whereas the HDL cholesterol level increased significantly. In a multivariate regression analysis, in addition to the initial risk-factor level, weight change was an important factor determining the changes in atherogenic risk factors. Changes in alcohol consumption were significantly associated with the changes in systolic blood pressure and HDL cholesterol levels. Changes in cigarettes smoking also showed significant associations with the changes in triglyceride level and the ratio of total cholesterol to HDL cholesterol. These results suggest that although the education program for controlling weight may have limited success, weight reduction exhibits beneficial changes in the atherogenic risk-factor profile in middle-aged obese men. PMID- 10812849 TI - Magnetometric evaluation for the effects of silicon carbide whiskers on alveolar macrophages. AB - Alveolar macrophages are thought to play an important role in fibrogenesis in the lungs caused by various types of exposure to dust. In this experiment, we evaluated the effect of silicon carbide whiskers (SiC) on alveolar macrophages mainly by unique magnetometry and also by established methods such as lactate dehydrogenase (LDH) activity, apoptosis measurement and morphological observations. Alveolar macrophages obtained from Syrian golden hamsters by bronchoalveolar lavages were exposed in vitro to Fe3O4 for 18 hours as an indicator for magnetometry and SiC for experiments. A rapid decrease of the remanent magnetic field, so called "relaxation", was observed after cessation of an external magnetic field in macrophages phagocytizing Fe3O4 alone, while relaxation was delayed in those concurrently exposed to SiC. Release of LDH from SiC-exposed macrophages into the medium was not significantly higher than the controls, but it increased dose-dependently. Apoptosis was recognized in macrophages exposed to 60 micrograms/ml of SiC by the DNA ladder detection method and morphological observations. Electron microscopic examination revealed irregular forms of nuclei and organellas in macrophages exposed to SiC. Magnetometry, LDH release and electron microscopic observation indicated mild cytotoxicity of SiC to alveolar macrophages. PMID- 10812850 TI - Does job stress affect injury due to labor accident in Japanese male and female blue-collar workers? AB - To clarify whether job stressors affecting injury due to labor accidents differ between Japanese male and female blue-collar workers, the Job Content Questionnaire (JCQ), assessing dimensions of job stressors based on the demand control-support model, was applied to 139 blue-collar workers in a manufacturing factory. Of them, 24 male and 15 female workers suffered from injuries at work. In the female workers with the experience of work injury, the job demand score and job strain index (i.e., the ratio of job demand to job control) of the JCQ were significantly higher and the score of coworker support was significantly lower, than those in the female workers without the experience. High job demand (or, high job strain and low coworker support) was significantly related to work injury in all the female workers. Between the male workers with and without work injury, however, there was no significant difference in any job stressors. This pilot study suggests that high job strain (specifically, high job demand), as well as low coworker support, are important factors affecting work injury in Japanese female blue-collar workers. Further research with a large number of male blue-collar workers will be required to seek other factors that may be associated with work injury. PMID- 10812851 TI - Evaluation of combined effect of organic solvents and noise by the upper limit of hearing. AB - To clarify the combined effects of organic solvents and noise on hearing, the upper limit of hearing was measured in 48 male workers exposed to organic solvents and/or noise in a factory producing plastic buttons. Measuring the organic solvent concentrations in working environments and breathing zone air, and the noise level in workplaces were also done. The readings suggested that most exposures to organic solvents and/or noise were within the occupational exposure limits. The prevalence rate of the upper limit of hearing below 75th percentile curve was higher in workers exposed to organic solvents and noise than expected (25 percent) and the other groups, whereas it was about 25 percent in the noise and control groups. The results suggest that the probable combined effects of organic solvents and noise on hearing should be considered even when the exposures are within the occupational exposure limits. PMID- 10812852 TI - Birth of a midwife. PMID- 10812853 TI - Clinician-specific episiotomy rates: impact on perineal outcomes. AB - Recent, large, randomized, controlled trials of the effects of episiotomy on perineal damage have confirmed that episiotomy is associated with an increased risk of damage to the perineum. Yet episiotomy remains the most common surgical procedure women undergo. This article examines if clinician experience, rather than scientific evidence, forms the basis for continuing this practice. Perineal outcome data are analyzed for 865 low-risk women who were attended at birth by the staff nurse-midwives or faculty obstetricians at a university-based, tertiary care hospital. Data were collected under routine, nonexperimental conditions such that the circumstances of the labor and the clinician's preferences were allowed to determine management decisions regarding the use of episiotomy or other techniques of perineal management. Multivariate findings indicate that in the absence of episiotomy, rates of perineal integrity were highest among clinicians who usually had the lowest rate of episiotomy use. When an episiotomy was done, rates of third- and fourth-degree extensions were highest among clinicians who used episiotomy most frequently. This finding challenges the idea that clinicians who were very experienced with the use of episiotomy would avoid complications such as extensions. Future research should explore the use of nonsurgical techniques such as those employed by midwives to promote perineal integrity. Then interdisciplinary research and evidence-based education regarding these techniques can occur to improve perineal outcomes for all women. PMID- 10812854 TI - Behavioral intervention for urinary incontinence in women: evidence for practice. AB - In the past, clinicians have relied heavily on pharmacologic and surgical interventions for urinary incontinence in women. However, evidence now exists that less invasive, behavioral therapies can be extremely effective in helping women become continent; thus, strategies that involve bladder and pelvic floor muscle training should generally be the first line of treatment. Before behavioral intervention is initiated, it is important to assess for any medical or associated conditions that should be treated first. Bladder training enables women to accommodate increasingly greater volumes of urine in the bladder and gradually to extend the interval between voiding. Pelvic floor muscle training increases awareness of function and strengthens these voluntary muscles, promoting continence. PMID- 10812855 TI - Placental triage of the singleton placenta. AB - Triage is the sorting and allocation of treatment according to a system of priorities in order to maximize treatment. Placental triage promptly after delivery of the placenta, with documentation of the findings in the medical record, only takes a few minutes, and allows for the identification of abnormal placentas to be submitted for detailed gross and microscopic pathology examination. It requires familiarity with normal gross placental anatomy and variations of normal, and the development of a systematic procedure for comprehensive examination of the placental disk as a whole, the umbilical cord, the extraplacental membranes, the fetal surface, the maternal surface, and the parenchyma. This article provides a systematic approach to gross examination of the placenta. It also reviews the clinical and legal impact of placenta examination, and suggests procedures that will optimize the information available in the placenta. PMID- 10812856 TI - Clinical decision-making regarding intravenous hydration in normal labor in a birth center setting. AB - Intravenous hydration (IVH) in normal labor is used routinely in most hospital settings despite the lack of support from recent research. However, it is not used routinely at one in-hospital, Alternative Birthing Center (ABC), in the Midwest. The purpose of this study was to explore and describe the criteria and perspectives of the certified nurse-midwives (CNMs) and nurses (RNs) in their decision-making process toward nonroutine use of IVH. In-depth interviews with five CNMs and four RNs that work in the ABC provided insight into how they balanced patient preference, research-based knowledge, and clinical judgments, in making decisions about IVH. The RNs were slightly more in favor of IVH than the CNMs, but not because of philosophic differences. Rather, the different roles and responsibilities of CNMs and RNs seemed to be most important. CNMs had a stronger commitment to client preferences, whereas RNs were more concerned about their responsibility for efficiently starting IVs prior to an emergency situation. Understanding professional differences in the decision-making process for IVH can enhance clinical functioning of the caregiving team by helping CNMs anticipate and address potential conflicts. PMID- 10812857 TI - Reducing perinatal transmission of HIV: early diagnosis and interventions during pregnancy. AB - This article reviews the New York State regulations regarding expedited testing of newborns for exposure to HIV. Included is a review of statistics as well as the medical and obstetric management of HIV positive pregnant women, including route of delivery. The professional responsibility of midwives, physicians, and other clinicians regarding maternal and neonatal health care is emphasized, especially in states without expedited testing. PMID- 10812858 TI - State regulation of midwives: issues and options. AB - In the United States, state governments play a central role in determining the extent to which midwives can provide care to women and babies. State laws and regulations establish midwives' scope of practice, set licensure requirements, and frequently determine their ability to get paid and obtain access to health care facilities. For certified nurse-midwives (CNMs), state regulation has evolved from a haphazard patchwork of highly variable regulatory models into a fairly uniform set of rules and requirements from one state to the next. For direct entry midwives (DEMs), there is much less uniformity, with some states outlawing practice by any midwife who is not a CNM, whereas other states have established rigorous standards and requirements for the licensure of DEMs. This article provides a broad overview of these state regulatory variables for both CNMs and DEMs, and explores issues and options that both state regulators and midwives should consider when developing or amending state laws and regulations governing their practice. In particular, the role of the state in regulating the practice of the certified midwife (CM) is examined in the context of existing regulatory frameworks for CNMs and DEMs. PMID- 10812859 TI - The 1999 ACC task analysis of nurse-midwifery/midwifery practice phase I: the instrument development study. AB - The national certification examination (NCE) in nurse-midwifery and midwifery is developed, administered, and evaluated by the ACNM Certification Council (ACC). The blueprint for the NCE is based upon a comprehensive list of tasks that describe the knowledge, skills, and abilities expected of the midwifery practitioner at entry into the profession. In 1999, the ACC initiated the third in a series of task analysis studies to ensure the currency and relevance of the task list. This study was considered particularly timely, given that the professional organization, the American College of Nurse-Midwives, had approved pathways to midwifery for individuals whose first degree was not in nursing (the certified midwife) and also had expanded the core competencies for midwifery practice to include responsibilities in the domain of primary care. This manuscript reports the results of the pilot study, in which the specific list of tasks was developed. Three hundred and six ACNM members responded to a preliminary list of tasks, indicating their opinion about whether each specific task was relevant to entry-level midwifery practice. The task list finally derived consists of 219 tasks and 177 clinical conditions, dispersed among seven domains of practice (antepartum, intrapartum, newborn, postpartum, well woman/gynecology, primary care/health assessment, and professional issues.) The task list represents a comprehensive profile of entry-level practice for nurse midwives and midwives certified by the ACC. PMID- 10812860 TI - Certified nurse-midwives' knowledge of reimbursement issues. AB - A recent survey examined the impact of reimbursement policies and admitting privileges on the ability of certified nurse-midwives (CNMs) to practice and maintain an income. Many CNMs reported deficits in their knowledge about reimbursement issues. The responses to the survey indicated that service directors either do not view this information as important to their leadership role, have delegated their authority to someone else, or do not have authority over this information. Education program directors may need to examine their curricular content to determine if students are being provided adequate information to better value and understand this data. PMID- 10812861 TI - Women's experiences of transfer from a midwife-led to a consultant-led maternity unit in the UK during late pregnancy and labor. AB - PURPOSE: This study explored the experiences of women who were transferred from a midwife-led to a distant consultant obstetric unit before or during labor. BACKGROUND: Little attention is given to the psychological impact of transfer, particular when it takes place prior to labor. METHOD: Narrative and progressively focused interviews were conducted with 18 women who faced or experienced transfer prior to or during labor. The data were analyzed using a grounded theory approach. RESULTS: The core category in the transferred group was loss. This related to loss of choice, control, continuity, and support and was associated with anger and resentment. Distress appeared most common when transfer took place late in a healthy pregnancy when the mother recognized no risk to the baby. CONCLUSIONS: More attention needs to be paid to the psychological impact of transfer from midwife-led to consultant-led care, particularly where this involves a change of location or midwife. PMID- 10812862 TI - From nursing outposts to contemporary midwifery in 20th century Canada. AB - Knowledge of the history of one's profession is always related to the informed practice of the profession and can aid midwifery in its expansion and recognition. Understanding the long and diverse history of Canadian midwifery expands the appreciation by midwives in the United States of midwifery north of our border, which, although newly regulated, has much experience to share. This article documents the evolution of midwifery in Canada during the 20th century. Government-sanctioned midwifery prior to 1991, the year when Ontario passed legislation to regulate midwifery for the first time in modern Canadian history, was limited to isolated, usually northern, frontier and outpost regions of Canada, but the providers who practiced it were well-educated and important figures in the broader history of midwifery. PMID- 10812863 TI - HIV in older Americans: an epidemiologic perspective. AB - The elderly population is increasing as baby boomers are beginning to approach retirement. People 65 years of age or older already constitute approximately one eighth of the U.S. population; this proportion is expected to double in the next 50 years. Older Americans have their own population-specific health challenges, such as Alzheimer's disease, osteoporosis, adult-onset diabetes, prostate cancer, menopause, and hypertension. Human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) are seldom discussed within this community. Prevention, counseling, testing, and education efforts are not being directed their way. In addition, few practitioners are experts both in HIV and health problems associated with aging, resulting in misdiagnosis, especially in the early stages when AIDS symptoms such as fatigue, weight loss, night sweats, and diminished appetite are dismissed as part of the aging process. Very few HIV related social support services have been aimed at the needs of the elderly, perhaps because older Americans are not suspected to be sexually active or are assumed to be in a monogamous, heterosexual relationship. Older Americans are not suspected of drug use. Yet many are sexually active, often demonstrating risky sexual behavior, such as dispensing with the use of condoms; and the isolation that frequently accompanies old age can lead to alcoholism and injectable drug use. This article examines methods suggested in the literature both in terms of primary and secondary prevention of HIV/AIDS in older Americans. The cost of these efforts is enumerated, and organizations who gear their efforts in reaching and educating older Americans regarding their risks are described. PMID- 10812864 TI - Characteristics of nurse-midwife patients and visits, 1991. PMID- 10812865 TI - The duration of labor in healthy women. PMID- 10812866 TI - Active phase labor arrest: oxytocin augmentation for at least 4 hours. PMID- 10812868 TI - Perspective. PMID- 10812867 TI - Effect of false-positive mammograms on interval breast cancer screening in a health maintenance organization. PMID- 10812869 TI - Finding the evidence for evidence-based dentistry. AB - Making sound clinical decisions about the proper use of materials, technology, and techniques in dental practice involves the correct interpretation of scientific data. Sources of such information include journals, textbooks, and the Internet. This article discusses the concept of evidence-based dentistry, the advantages and disadvantages of using the various media, particularly journals, for obtaining information, and how to judge the accuracy of what is presented. PMID- 10812870 TI - Evidence-based versus experience-based decision making in clinical dentistry. AB - Evidence-based dentistry has become a frequent topic of conversation because good practice must be based on good evidence. This article explores some of the characteristics of "best" evidence. It is concluded that a blending of sound science and practice experience is needed and it is proposed that such evidence can be generated in clinical study groups and the recently funded Centers of Discovery. PMID- 10812871 TI - CE institute style. AB - This article describes the philosophy and operation of The Pankey Institute for Advanced Dental Education as one example of the "institute" approach to professional development for dentists. PMID- 10812872 TI - Postgraduate educational opportunities in the military services. AB - The three branches of the service provide advanced training to about 450 dentists each year. Approximately one-third of these positions are in post-doctoral general dentistry and the remainder in specialties, distributed as needed by the various services. PMID- 10812873 TI - Professional growth, learning, and the study club. AB - The imperative for human growth and the need for continuous professional development are related. This article describes some of the practical decisions about forming a study club and finding a leader that promotes professional growth. Thoughts are offered about learning in this context as contrasted to school learning and about the psychology of growth. PMID- 10812874 TI - Dental ethics as an intellectual discipline: taking the next step. AB - The development of ethics education in dental and dental hygiene schools is traced. In the early 1980s curricula were informal, often incorporated with jurisprudence or practice management courses, and there were few effective educational materials. Over the next decade formal guidelines for teaching ethics had been approved by the American Association of Dental Schools, several texts and journals featuring ethics columns had appeared and PEDNET, the Professional Ethics in Dentistry Network had been created. The next step is to develop a regular forum for publication of papers in ethics in dental care. PMID- 10812875 TI - Commitment to community service: the story of Dr. Jack Echternacht. AB - Using the criteria developed by Colby and Damon, the authors identified Dr. Jack Echternacht as a moral hero. This paper describes his thirty-year fight to fluoridate the drinking water of Brainerd, Minnesota. Family influences on his character are also presented. PMID- 10812876 TI - Emerging trends in professional development. AB - The shape of professional development in dentistry can better be guessed from imagining the emerging needs of practitioners than from the technology of education. Four predictions are offered: professional development is likely to involve a greater blend of learning and doing, customized to individual dentists, emphasizing value added rather than evaluation of dentists, and to be distributed in the sense that dentists will take their learning in smaller doses and at their own convenience. PMID- 10812877 TI - Treating pain: new guidelines for using controlled substances. PMID- 10812878 TI - Complications of cystic fibrosis in adults. PMID- 10812879 TI - Questions and answers about organ and tissue donation. PMID- 10812881 TI - Ethnic disparity in health care--where are we now? PMID- 10812880 TI - South Carolina and Hill-Burton. PMID- 10812882 TI - Health care coverage of high school athletes in South Carolina. PMID- 10812883 TI - Study of lymphatic metastatic patterns in gastric carcinoma according to tumor location. AB - A total of 226 cases of advanced gastric cancer which occupied only one third of the stomach were analyzed in order to clarify whether and how lymphatic spread differed according to the tumor location and gross type of tumor. Out of the 226 patients, 45 cases had tumor in the upper third, 74 cases had it in the middle third, and 107 cases had it in the lower third of the stomach. The incidence of lymph node metastasis was found to be much higher for the tumors located in the lesser curvature (51.6%), greater curvature and posterior wall (54.4%), as compared to the tumors located in the anterior wall (28.0%). The tumors located in the upper third of the stomach did not show any metastasis in the N3 node, while the tumors located in the lower third of the stomach did not show any metastasis in the left cardial nodes, short gastric nodes, and the nodes along the left gastroepiploic vessels. Similarly, the tumors from the middle third of the stomach did not invade the left cardial nodes. PMID- 10812884 TI - gamma delta T lymphocytes and their V gamma 9 and V delta 2 receptor expression in peripheral blood mononuclear cells of active tuberculosis patients before and after treatment. AB - It has been reported that gamma delta T cells are activated by bacterial infection, and the cells act in an antibacterial manner. We investigated the immunologic role of gamma delta T cells and their receptors in tuberculosis (TB) patients. We examined gamma delta T cells which express receptors composed of V gamma 9 or V delta 2 chains before and after anti-TB chemotherapy, in vitro changes in T cell receptor expression due to stimulation, and the relationship between the proliferative capability of gamma delta T cells and the clinical data before treatment (10 TB patients and 9 healthy volunteers). The ratio of V gamma 9-positive cells to gamma delta T cells decreased significantly in the 10 TB patients before treatment (p < 0.05). In the in vitro study, the patients with a high pretreatment growth index of V delta 9-positive cells had a significantly increased erythrocyte sedimentation rate (p < 0.05) and significantly low rates of V gamma 9- and V delta 2-positive gamma delta T cells (p < 0.02). Measurement of the proportion of V gamma 9-positive cells together with their growth index might help to further elucidate the disease process in patients with pulmonary TB. PMID- 10812885 TI - The effect of allogeneic or xenogeneic immune responses and preservation techniques on transplanted aortic valve grafts. AB - We examined the effect of allogeneic and xenogeneic immune responses on the histopathological changes in aortic valve grafts and the influence of preservation techniques on these changes. Brown Norway rats and Syrian hamsters were used as allogeneic and concordant xenogeneic donors of aortic valve grafts, respectively. The allografts and xenografts were implanted heterotopically in the abdominal aorta of Lewis rat recipients immediately after harvest (homovital), after cryopreservation, or after preservation with antibiotics at 4 degrees C (fresh preservation). Allografts and xenografts were explanted at days 7, 28 or 56 and at days 3, 7 or 14, respectively, for the histopathological examination. The allografts underwent histological changes characteristic of graft arteriosclerosis. No significant effect of cryopreservation on these changes was observed. The fresh-preserved graft was, however, predisposed to focal destruction of the elastic fibers and to early disappearance of the leaflet. The lesions in xenografts were characterized by severe destruction of the elastic fibers. Compared to homovital xenografts, both cryopreserved and fresh-preserved xenografts showed more prominent disruption of the elastic fibers, well-developed valvular and vascular thrombi and earlier disappearance of the leaflet. In conclusion, it could be assumed that failure in retention of cellular and extracellular components during fresh preservation accelerates structural deterioration of allografts. As for xenografts, even the extracellular matrix may have potential xenogeneic immunogenicity. There is a possibility of these preservation techniques reducing xenogeneic immunogenicity of the endothelial cells, probably because of loss of these cells. However, it appears that, even in this setting, other cellular and extracellular components could trigger immune responses causing structural deterioration of xenografts. PMID- 10812886 TI - Epidemiological study on food intake and Helicobacter pylori infection. AB - We conducted an epidemiological study to investigate the relation of food intake to Helicobacter pylori (H. pylori) infection in an area endemic for H. pylori. In this study, 365 subjects, 104 men and 261 women, were randomly selected from 7,389 adult (over age 20) inhabitants of town A, Japan. The prevalence of immunoglobulin G (IgG) class antibody to H. pylori (anti-H. pylori) was 83.7% and the prevalence of anti-H. pylori increased with age significantly (P < 0.05). Subjects with anamnesis of gastritis, gastroduodenal ulcer and gastric cancer tended to have a higher anti-H. pylori positive ratio (93.5%) than those without (81.0%). But there was no relationship between anti-H. pylori prevalence and sex, blood type, smoking or drinking habits. Daily intake of foods by food groups, nutrients and the concentrations of serum ingredients were compared between 37 anti-H. pylori-positive and 40 negative subjects selected from 365 inhabitants by matching up according to sex and age. The daily intake of cereals, potatoes and starches, and milks tended to be higher in positive than negative subjects, while the daily intake of algae and tea appeared to be a little higher in negative than in positive subjects. The daily zinc intake of antibody-positive subjects was significantly higher (P < 0.05) than in antibody negative subjects. On the other hand, the daily iron intake in negative subjects was significantly higher (P < 0.05) than in positive subjects. The serum concentrations of copper, zinc, and vitamin E tended to be higher in positive than negative subjects. But there were no significant differences in serum ingredients concentrations between antibody negative and positive subjects. Our findings suggest that iron and zinc intakes may effect on H. pylori infection. PMID- 10812887 TI - Morphological change in the MNNG-treated rat gastric mucosa. AB - To investigate the process of carcinogenesis in gastric cancer, we studied the histological features of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-treated rats. Samples of the gastric mucosa from both MNNG-treated and control rats were histologically examined every 2 months, for 10 months. In 40% of the MNNG-treated rats, atrophy in the gastric mucosa was observed after 2 months, and regenerative epithelium was observed after 4 months, followed by adenomatous proliferation and disappearance of the tight junction electron microscopically after 6 months. A small intestinal cancer had developed in 2 rats at 6 months. While gastric cancer had developed in 3 rats at 8 months, and in one of these 3 rats, peritoneal dissemination was observed macroscopically and histologically. These results suggested that adenomatous proliferation and disappearance of the tight junction observed electron microscopically were characteristic pathological features of precancerous lesions in the stomach in MNNG-treated rat. PMID- 10812888 TI - Idiopathic osteonecrosis of the femoral head in the elderly. AB - Idiopathic osteonecrosis of the femoral head (ION) is a disease prevalent in adulthood, and its development in elderly persons is known to show a low incidence. To clarify its characteristics in elderly patients, radiological and histological examinations were performed on 18 patients aged 60 years or older histologically diagnosed as having ION. On roentgenography, three characteristic points were observed, 1) progress of collapse in the femoral head within a short period (12 cases, 66.7%), 2) narrowing in the joint space within a short period (6 cases, 33.3%), and 3) rapid destruction and resorption of the femoral head (4 cases, 22.2%). The lesion expanded to the acetabular region, and a course similar to that of so-called rapidly destructive coxarthrosis (RDC) was developing in 3 cases (16.7%). However, in these 3 cases, changes in the acetabular region were mild, compared with ordinary RDC, and they were considered different from common RDC. These radiological characteristics were found more clearly in women than in men. In the histopathological examination, severe degeneration in the cartilage of the femoral head was found, while wide infiltration of repairing tissue into the necrotic area was seen from the early stage. When necrotic trabecula were collapsed markedly and degraded severely, then destruction and resorption of bone expanded to non-necrotic regions in some cases. Osteogenic sclerotic line (the demarcation line), which has been often seen in ordinary ION, was not developed in most cases. These characteristics were more prominent in women. PMID- 10812889 TI - Spinal cord protection: effect of N-methyl-D-aspartate receptor antagonist MK-801 for spinal cord ischemia in a rabbit model. AB - Excitatory amino acids (glutamate, aspartate) play an important role in the ischemic cascade leading to cell death. The N-methyl-D-aspartate (NMDA) receptor is an excitatory amino acid (EAA) receptor, and NMDA receptor antagonists have been shown to exert a neuroprotective effect in central nervous system ischemia. The purpose of this study was to investigate the effects of noncompetitive NMDA receptor antagonists MK-801 and to observe the changes in EAAs after spinal cord ischemia in a rabbit model. Spinal cord ischemia was induced by clamping the infrarenal abdominal aorta for 24 min. Group 1 (n = 6) received no pharmacologic infusion. Group 2 (n = 5) was administered an intra-aortic hypothermic MK-801 (1 mg/kg) solution and group 3 (n = 6) was administered an intra-aortic normothermic MK-801 (2 mg/kg) solution immediately after clamping of the abdominal aorta. We evaluated the neurological function at 12, 24 and 48 hrs after spinal cord ischemia. A histopathologic study was carried out 72 hrs after spinal cord ischemia, and the results for groups 1 and 3 were compared. The glutamate and aspartate levels in the blood plasma were compared at pre-ischemia and at 12, 24, and 48 hrs among the groups. The perfusion of a normothermic MK-801 (2 mg/kg) solution significantly reduced the neurological dysfunction and the neuronal damage. There was a significant increase in aspartate at 24 and 48 hrs in group 1, but no such increase in glutamate occurred in groups 1 and 3. In conclusion, these data provide the evidence that therapeutic intervention with MK-801 (2 mg/kg) in the early period of spinal cord ischemia is beneficial in reducing neurological dysfunction and neuronal damage. PMID- 10812890 TI - Energy metabolism after cardiac surgery utilizing the indirect calorimeter: a new breath by breath technique. AB - The aim of this study is to clarify the relationship between pyrexia and energy expenditure (EE) after cardiac surgery utilizing the indirect calorimeter. Further the present study tried to determine in changes of energy substrate during mechanical ventilation after cardiac surgery. EE, oxygen consumption (VO2), carbon dioxide production (VCO2), cardiac index (CI), core temperature, and energy substrate were continuously measured in 26 patients after cardiac surgery. The patients were divided into two groups according to the highest core temperature postoperatively; Pyrexia group: > or = 38.5 degrees C, Non-pyrexia group: < 38.5 degrees C. EE after cardiac surgery in non-pyrexias was 1.33 times higher than calculated EE according to the Harris-Benedict equation. Moreover, EE in pyrexias was approximately 1.6 times higher. The mean EE in pyrexias increased to 27.8%/degree C. No significant differences were observed in VO2 or VCO2 between two groups. Time course of CI in pyrexias was different from that in non pyrexias. Accordingly, energy substrate was not shifted in both groups except fat. This study suggested that the maintenance of core temperature below 38 degrees C after cardiac surgery is important on the aspect of energy metabolism. PMID- 10812891 TI - Effects of LTB4 receptor antagonist on myonephropathic metabolic syndrome: an experimental study. AB - The aims of this study were to determine the involvement of leukocytes in reperfusion injury following acute arterial occlusion and to evaluate the effect of the leukotriene B4 (LTB4), which is a chemical mediator of inflammation, receptor antagonist. We examined the usefulness of LTB4 receptor antagonist, ONO 4057, as a preventative drug for myonephropathic metabolic syndrome (MNMS). The experimental leg ischemic model was developed using Wistar strain rats. The rats were divided into 4 groups. In Group R3, the infra-renal abdominal aorta was clamped for 3 hrs and the right femoral muscle tissue was cut to block the development of a collateral artery. In Group R6, the infra-renal abdominal aorta was clamped for 6 hrs and the right femoral muscle tissue was cut. In Group C, the controls, there was no clamping of the abdominal aorta and the right femoral muscle tissue was cut. In Group M, the medicated group, rats were pretreated with an LTB4 receptor antagonist, ONO-4057, just before reperfusion. Blood serum interleukin-1 (IL-1), interleukin-8 (IL-8), creatine phosphokinase (CPK), and aldolase were measured and compared in each of those 4 groups. We also examined the intercellular adhesion molecule-1 (ICAM-1) expression in various organs (liver, heart and kidney) by immunohistochemistry. We found that IL-1 beta levels were low in all groups. CPK, aldolase and IL-8 levels after reperfusion in Group R6 significantly high compared with the levels in Group C (P < 0.03 about CPK, P < 0.05 about aldolase, and P < 0.05 about IL-8). The levels of CPK, aldolase, and IL-8 in Group M were significantly lower than those in Group R6 (P < 0.02 about CPK, P < 0.04 about aldolase, and P < 0.03 about IL-8). We determined immunohistochemically that the expression of ICAM-1 was positive on endothelial cells at the coronary artery and the small vein in Group R6 and that the expression of ICAM-1 was negative on endothelial cells in Group C. Those data suggested that ICAM-1 may play an important role in the progression of reperfusion injury, and the adhesion of neutrophilic leukocytes on endothelial cells may play a significant role in MNMS. LTB4 receptor antagonist may be useful for preventing reperfusion injury following acute aortic occlusion. PMID- 10812892 TI - Effects of holmium(HO)-YAG laser irradiation on rabbit lumbar discs. AB - To study the effect of laser irradiation on normal lumbar discs, a 2100 nm Holmium (HO)-YAG laser irradiation was applied to the 83 lumbar discs of 23 adult rabbits. The extent of disc vaporization, the temperature changes in the surrounding tissues, and changes in the radiograph and MRI findings were assessed after laser irradiation. When laser irradiation was delivered to the discs, the disc weight decreased linearly with the increase in total laser energy, indicating steady vaporization of disc material. The temperature was highest at the site of the guide needle. Laser irradiation was delivered at 0.5 J/pulse or 1.4 J/pulse x 5 pulses/sec to the intervertebral discs, and radiographs and T2 weighted MRI of the irradiated discs were investigated at 1, 4, and at 12 weeks after irradiation. At 1 week after irradiation at 0.5 and 1.4 J/pulse, the radiographs showed a decrease in the disc height. At 12 weeks after irradiation at 0.5 J/pulse, the disc height had restored to normal, while the decrease was persistent after irradiation at 1.4 J/pulse. At 1 week after irradiation, MRI showed a decrease in the signal intensity of discs treated at 0.5 J/pulse, but the decrease was recovered at 12 weeks. After irradiation at 1.4 J/pulse, the decrease in signal intensity was also recovered by 12 weeks, but the recovery was less than the recovery after treatment at 0.5 J/pulse. Laser irradiation is applicable for the treatment of intervertebral discs, but it is necessary to select the optimal operating conditions. It may also be necessary to change the power of irradiation according to the pathological condition of the disc being treated. PMID- 10812893 TI - Adverse effects of whole-body vibration on gastric motility. AB - To investigate the response of gastric motility to whole-body vibration (WBV) exposure, electrogastrography (EGG) and gastric manometry were performed in 10 healthy male volunteers. Sinusoidal vertical vibration of three different frequencies (4 Hz, 8 Hz, and 16 Hz) with a constant vibration magnitude of 1.0 ms 2 (rms.) was randomly given to the subject seated on the platform of a vibrator for 10 min. Exposure to vibration of 4 and 8 Hz decreased the amplitude of EGG wave and of the power spectrum corresponding to a slow wave component at fasting state. Food intake (solid meal 80 g, 135 cm3, 400 kcal) enhanced gastric motility showing about 2.5-fold in the power spectrum, of which response modes during and after vibration exposure were similar to those at fasting state. The periodical manometric change around one cpm was observed during vibration exposure under the condition of food intake. Short-term exposure to WBV led to a suppression of the activity of gastric smooth muscles and affect contraction wave. These responses may result from resonance of vibration frequency as a mechanical factor and stomach contents, and increase regulation of neurohumoral factors due to vibration stress. PMID- 10812894 TI - Gastric stromal tumor with CD34 immunoreactivity--a case report. AB - Gastric stromal tumors are the most common mesenchymal tumors, and such submucosal mass lesions of the upper gastrointestinal tract occur frequently. A 54-year-old woman with no major complaint was admitted to our hospital for evaluation of a mass located between the stomach and the pancreas. Abdominal ultrasonography, computed tomography and endoscopic ultrasonography demonstrated a mass lesion which was located near the lesser curvature of the stomach. Selective left gastric arterial angiography revealed a hypervascular mass, and we diagnosed it as a leiomyosarcoma of the stomach. At laparotomy, there was a large solid mass 5 cm in diameter along the minor curvature of the stomach. Tumor resection with partial gastrectomy was performed, and the histological diagnosis was a gastric stromal tumor with CD34 immunoreactivity. We report a case of stromal tumor of the stomach with extramural growth and review the literature. PMID- 10812895 TI - Successful removal of an infected pacemaker electrode by open heart surgery under extracorporeal circulation. AB - A 33-year-old male with sick sinus syndrome, who had received a pacemaker implant 18 years earlier, was complicated with a generator infection. Although the infected generator was removed, he was suffered from the recurrent local infection associated with a retained pacemaker lead. After a new pacemaker system implantation from the other side of the subclavian vein, we attempted to remove the lead utilizing a pacemaker removal kit. However, this intervention procedure was unsuccessful, because fibrous adhesions had developed around the lead, accompanied by calcification along its course. As a last resort, we opened the heart under extracorporeal circulation and removed the lead under direct vision. The post-operative course was uneventful. In order to remove a long-term implanted pacemaker lead, the direct surgical procedure with extracorporeal circulation is a favorable mean alternative to conventional intervention techniques. PMID- 10812896 TI - HBV DNA can be detected from nail clippings of HBs Ag positive patients. PMID- 10812897 TI - [Monopoly]. PMID- 10812898 TI - [Urea in dermatology]. PMID- 10812899 TI - [Drug therapy of Parkinson disease]. PMID- 10812900 TI - [Headaches in otorhinolaryngology]. PMID- 10812901 TI - Judicial oversight of the nation's largest guardianship system: caselaw on Social Security Administration representative payee issues. PMID- 10812902 TI - Prostate cancer: correlation between local and systemic disease. AB - BACKGROUND: Prostate cancer is in Italian men the second neoplasm for incidence. Transrectal ultrasonography (TRUS) is still today a cheap test, not very invasive, well accepted by the motivated patient and sufficiently accurate for the valuation of the local status disease. We proposed our data to verify if a careful local ultrasonographic study of prostatic carcinoma could or not predict its systemic course. METHODS: We have valued 136 out patients affected by prostate cancer and treated with either palliative medical therapy or radiotherapy. The follow-up varies from 12 to 132 months. The local, evaluation of the disease done through TRUS in longitudinal and/or axial scan (the dimension of the gland, the volume and ultrasonographic characteristics of the lesion, capsular involvement and estimation of the surrounding structures: seminal vesicles, vesica, rectum). RESULTS: In the long run, considering the local and systemic course of the disease, we have noticed accordance globally in 116 patients (85.3%); 13 patients showed only systematic progression (9.5%) while in 7 patients, we have noticed a local but not a systemic variation. CONCLUSIONS: We can deduce that periodical carrying-out of the TRUS (twice a year) constitute a reliable index not only of local procedure of the pt. but also predicting the systemic course of the disease, making the routine carrying out of the other check-up unnecessary, that could be reserved for cases with precise clinical indication. PMID- 10812903 TI - Treatment of benign prostatic hyperplasia with transurethral electrovaporization of the prostate (TUVP) using Vaportrode VE-B. Two years follow-up. AB - BACKGROUND: The authors report their experience on transurethral vaporization of the prostate with Vaportode for the treatment of BPH. METHODS: In January 1996 we treated 24 patients suffering from cervico-urethral obstruction caused by BPH with electrovaporization of the prostate using a cutting current of between 180 and 250 Watts (average 200 Watts). Exclusion criteria was: PSA > 3 ng/ml. The catheter was removed after 24 hours. Follow-up of all patients is 27 months. Twenty-three patients were evaluated. RESULTS: Results at follow-up were; mean maximum flow (Qmax) 9.31 ml/sec preoperative was 19.5 ml/sec after 6 months, 18.3 after 12 months and 18.0 after 24 months; mean symptom score decreased from 18.1 to 10.4 after 6 months, to 3.2 after 12 months and to 4.8 after 24 months; QL index decreased from 4.6 to 2.3 at 6 months, to 1.4 after 12 months and 24 months; the average avoiding pressure decreased from 62.8 cm/H2O to 23.2 cm/H2O after 6 months, to 21.2 after 12 months and to 23.8 after 24 months. There was no post voiding residual urine 2 days after treatment in all patients. CONCLUSIONS: Follow-up shows transurethral electrovaporization of the prostate is an efficient and safe technique for treatment of BPH. PMID- 10812904 TI - [TB-test in the diagnosis of "closed" tuberculosis of the urinary tract. Preliminary study]. AB - BACKGROUND: Tuberculosis (TBC) is a disease that is found throughout the world with a particularly high incidence in under developed countries. Genitourinary tuberculosis has been reported in 8-10% of all cases in developed countries and in 20% in Third World countries. This study aimed to describe a method of diagnosis applicable to so-called "closed" forms of tuberculosis affecting the urinary tract, namely those with negative results for KB bacteriological and culture assay, but in which suspected diagnosis based on clinical grounds and diagnostic procedures call for further analysis; these forms account for 10% of the total cases. METHODS: A prospective study was carried out in order to tritade the antigenic fraction A60 of the group of thermostable macromolecular antigens (TMA); the latter belong to the pool of liposoluble antigens forming the cell wall and represent the main component of tuberculin and purified protein derivative (PPD). The identification and assay of antigen A60, which can stimulate both the antibody and cell-mediated response, was performed using the ELISA method in order to identify IgG and IgM specific antibodies (TB-test). Twenty-nine patients were selected (11 males, 18 females) with a clinical history of recurrent symptomatic manifestations of upper and lower urinary tract infections resistant to common antibiotic treatment. Urinary tests showed that 96.5% of patients presented an acid urinary pH associated with pyuria, and the urographic imaging revealed calico/pyelic lesions of the excretory tracts. RESULTS: Seven patients (24%) showed a form of urinary TBC of an areactive type (AA) or with intermediate reactivity (RI.AI); six patients (20.64%) showed reactive TBC or in the process of resolution; fifteen patients (51.6%) were negative to the TB-test, and after one year's follow-up still showed no positivity to assays for Koch's bacillus. CONCLUSIONS: The authors affirm that the TB-test, a rapid and relatively inexpensive diagnostic method, is a valid aid for the diagnosis of "closed" forms of urinary tract TBC. PMID- 10812905 TI - [Chronic renal insufficiency and growth retardation: not an inseparable binomial]. AB - BACKGROUND: The aim of the study was to evaluate, in children with chronic renal failure and in conservative treatment, the degree to which their growth was compromised, in relation with bone age and with GRF. METHODS: The growth of children (12 F, 14 M) with CRI has been evaluated retrospectively in relation to bone age and GFR. Their mean age at diagnosis was 5.8 years. The pathologies causing CRI were, in particular, congenital anomalies and hereditary nephropathies. The mean follow-up was 6.1 years. RESULTS: No significant difference was found between bone age (BA) and chronologic age (CA) (4.7 +/- 3.4 vs 5.7 +/- 3.5 years, p < 0.3) at diagnosis, while it was lightly delayed (8.6 +/ 5.2 vs 11 +/- 5.6 years, p < 0.07) at the end of the study. At diagnosis the mean height SDS for CA was 0.64 +/- 1.12, at the end of observation was -1.2 +/- 1. Only in 3 patients height was < -2DS: two of them underwent rhGH treatment; the third patient had a growth potential exhausted. The mean height velocity (HV) was 6 +/- 4.2 cm/y at diagnosis and 4.8 +/- 3 at last observation (p < 0.3). No significant difference at diagnosis was found between the HV cm/y and GRF stratified for range, at follow-up there was light significant difference between HV and light CRF vs severe (p < 0.02). Likewise no significant relationship between HV and CA was observed. Actually 20 patients are in conservative treatment, 6 received renal transplantation: for them the growth has been proportional to reached renal function and age of receiver. CONCLUSIONS: The conservative therapeutic regimens and good compliance allowed to limit growth deficiency. PMID- 10812906 TI - [Arteriovenous fistulas for hemodialysis: transposition of the cephalic veins, a personal experience]. AB - BACKGROUND: Vascular access for hemodialysis has remarkably developed during these years. Since 1966 we have the Brescia-Cimino fistula which is considered nowadays the best choice for angioaccess. The transposed cephalic vein is the "variant" which has been evaluated in a single stage surgical technique. METHODS: A prospective and randomized study regarding 23 patients submitted to operation for first fistula during 1998. With a median follow-up of 10 months, these fistulas have been studied with echocolordoppler in order to verify their primary patency, diameter and blood flow in artery, in vein and anastomosis. RESULTS: Fistulas have been patented in all cases (15 males e 8 females) and used for dialysis after 3 weeks. No early or late complications have been observed. Mean diameter has been 1.2 cm with mean velocity of 1.8 m/sec, in artery mean velocity 2.3 m/sec and in vein 1.1 m/sec. CONCLUSIONS: Increasing length of life even for high critical patients like these with chronic renal failure underlines the need of surgical strategies which may improve the quality to life. This technique of transposed cephalic vein has the same advantages of direct fistulas also for those patients in which we should have used prosthetic grafts. PMID- 10812907 TI - [Frequency and antibiotic susceptibility of urinary pathogens isolated in the microbiology and virology laboratory of the Careggi General Hospital in the period July-December 1996]. AB - BACKGROUND: The aim of the study was to evaluate the isolation frequency and antibiotic susceptibility of pathogens isolated form urinocoltures. METHODS: 2192 bacterial strains obtained from urine cultures with bacterial count > 100,000 UFC/ml were examined in our Laboratory from July through December 1996. Inpatients as well as outpatients were considered. Five different ward typologies were taken into account (Surgery, Medicine, Obstetrics, Spinal Unit and High Risk Wards). Isolation frequencies were evaluated for each ward, but in order to get an adequate statistical sample they were divided into in two categories, e.g. strains isolated from inpatients and from outpatients. Antibiotic susceptibility was also evaluated dividing the data into the same two categories. For strains isolated with lower frequencies that was not possible, and data from inpatients were grouped with data from outpatients. RESULTS: The results obtained show that bacterial species most frequently isolated among inpatients are Escherichia coli (45.7%), Enterococcus faecalis (16.8%), Proteus mirabilis (9.1%), Pseudomonas aeruginosa (6.9%) and Klebsiella pneumoniae (6.4%). Those most frequently isolated among outpatients are Escherichia coli (58.1%), Enterococcus faecalis (12.3%), Proteus mirabilis (9.1%), Klebsiella pneumoniae (4.4%) and Coagulase negative staphylococci (4.0%). As far as antibiotic susceptibility is concerned, whenever a statistically significant difference in sensitivity could be observed between strains isolated from inpatients and strains isolated from outpatients, sensitivity was always higher for outpatients strains. CONCLUSIONS: Isolation frequency and antibiotic susceptibility are different depending on the place of origin of the patients (inpatients/outpatients). This implies a different approach to the empiric therapy in urinary tract infections. PMID- 10812908 TI - [Fibrous pseudotumors of the tunica vaginalis testis. A clinical case]. AB - A case of multiple fibrous pseudotumors of the tunica vaginalis is reported. Ultrasonography led to diagnosis 10 years previously and the patient underwent surgical exploration when the lesions were up to 6 cm in diameter. PMID- 10812909 TI - [Retroperitoneal mixoid liposarcoma: a difficult diagnosis with often unfavorable prognosis. Case report]. AB - Liposarcoma is the most common adult soft tissue sarcoma. The hallmark is the immature fat cell or lipoblast. It occurs in late adult life, frequently in the lower extremities, in the retroperitoneal, perineal and mesenteric region. Retroperitoneal liposarcoma doesn't produce symptoms until it is very large. A review of the literature is made and the case of a 65-years old man, submitted to surgical treatment for a bulky retroperitoneal mixoid liposarcoma is reported. PMID- 10812910 TI - Births, marriages, divorces, and deaths: provisional data for April 1999. PMID- 10812911 TI - A talk with Irving Ratner, MD. 207th president of MSNJ. Interview by Leah Z. Ziskin. PMID- 10812912 TI - Physician's profile helps patients and doctors. PMID- 10812914 TI - Looking toward the future: MSNJ vice presidents predict conference impact. PMID- 10812913 TI - Focus on the Academy of Medicine: commitment to education and unity. PMID- 10812915 TI - New Jersey prepares for a possible return of West Nile virus. PMID- 10812916 TI - Disability awareness: a checklist for the medical practice. AB - Although Congress passed the Americans with Disabilities Act (ADA) of 1990, our nation's 54 million people with disabilities continue to face many challenges when seeking health care. Physicians and their staffs can heighten their awareness and sensitivity to disability and provide better communication plus an organized system of care for patients with disabilities. PMID- 10812917 TI - Practice consolidation and retirement plans: financial ramifications. PMID- 10812918 TI - Appellate court upholds MIIX conversion. AB - On February 14, 2000, the Appellate Division of New Jersey Superior Court issued an extensive opinion that firmly upheld MIIX's conversion from a reciprocal to a stock insurance company. The court ruled that the MIIX plan of reorganization, by which the conversion was accomplished, was carried out strictly in accordance with law and that the plan's method of distributing stock in the new company to MIIX's members was fair and reasonable. This decision conclusively rejected the challenge to the plan made by three dissident MIIX member physicians. PMID- 10812919 TI - Energetic adaptation to chronic disease in the elderly. AB - Several chronic diseases occur with increased prevalence in the elderly. Body weight loss is a common feature of many chronic diseases. Weight loss increases the risk for morbidity and mortality and contributes to decreased functional independence and poor quality of life. Thus, an understanding of the effect of chronic disease on energy balance has important implications for nutritional supplementation and clinical outcome. This brief review will consider recent studies that have examined the effect of several chronic diseases (i.e., Alzheimer's disease, Parkinson's disease, and congestive heart failure) on daily energy expenditure in elderly individuals. Additionally, we put forth a model to explain the energetic adaptation to chronic disease in the elderly that is based on measurements of daily energy expenditure and its components. Studies suggest that chronic disease decreases daily energy expenditure in elderly individuals due to a marked reduction in physical activity energy expenditure. Moreover, these changes in daily energy expenditure often occur in the presence of increased resting energy expenditure. Thus, the net effect of chronic disease is to decrease daily energy expenditure. These results do not favor the hypothesis that increased energy expenditure contributes to disease-related weight loss. Instead, reduced energy intake appears to be a more likely mediator of the negative energy imbalance and weight loss that frequently accompany chronic disease in the elderly. PMID- 10812920 TI - Quest for the molecular mechanism of chromium action and its relationship to diabetes. AB - Despite forty years of research on the potential role of chromium in carbohydrate and lipid metabolism, significant progress has only recently been made regarding the mode of action of chromium at a molecular level. The oligopeptide low molecular-weight chromium-binding substance (LMWCr) may function as part of a novel insulin-signaling autoamplification mechanism. The proposed mechanism of action also sheds some light on the potential of chromium-containing compounds as nutritional supplements or in the treatment of adult-onset diabetes and other conditions. The potential relationship between the results of recent studies on diabetic patients and the proposed mode of action of LMWCr are discussed. PMID- 10812921 TI - Does vitamin C intake protect against lead toxicity? AB - Although several animal studies suggest a protective relationship between blood lead concentrations and ascorbic acid, there are inconclusive results regarding the beneficial effect of ascorbic acid on lead concentrations in human studies. Data from the Third National Health and Nutrition Examination Survey examined the association between ascorbic acid and blood lead concentrations in 19,578 participants ages 6-90 years without a history of lead poisoning. Elevated blood lead concentrations were found in 0.4% of adults and 0.5% of youths. Serum ascorbic acid concentrations were inversely associated with the prevalence of elevated blood lead concentrations. However, there was no significant relationship between dietary ascorbic acid intake and blood lead concentrations. This study suggests that there may be a protective relationship between ascorbic acid and lead. Questions remain regarding the unique roles of dietary vitamin C versus supplemental vitamin C in explaining this relationship. PMID- 10812922 TI - ABC1: the gene for Tangier disease and beyond. AB - Coronary heart disease (CHD) is the leading cause of death in America. CHD is multifactorial, and low plasma high-density lipoprotein cholesterol (HDL-C) levels are among the most common biochemical abnormalities observed in CHD patients. The mechanisms controlling plasma HDL-C levels are poorly understood. However, several groups recently reported that mutations at the ATP-binding cassette transporter 1 gene (ABC1) are responsible for a rare disorder known as Tangier disease, which is characterized in the homozygous state by the virtual absence of circulating plasma HDL. This new finding represents a major breakthrough in our knowledge of lipoprotein metabolism and, more specifically, the reverse cholesterol transport. This information could lead to a more precise assessment of the genetic predisposition to CHD as well as to new therapeutic tools to prevent and treat CHD. PMID- 10812923 TI - Oral rehydration solution therapy in the management of children with rotavirus diarrhea. AB - Rotavirus infections are the most common cause of gastroenteritis among children younger than 3 years of age and are associated with sporadic outbreaks of diarrhea in elderly and immunocompromised patients. Oral rehydration solutions (ORS) are formulated to correct dehydration and acidosis. Currently, ORS do not promote intestinal healing; however, investigators are examining the role of nutrition in promoting intestinal healing. This article reviews the composition of several ORS in human medicine and summarizes our current knowledge of the nutritional treatment of rotavirus diarrhea and intestinal healing. PMID- 10812924 TI - Tea oxalate. PMID- 10812925 TI - Overview: what is insulin resistance? PMID- 10812926 TI - Epidemiology. PMID- 10812927 TI - Role of dietary factors: macronutrients. AB - Insulin resistance is an important early marker of the metabolic syndrome disease cluster. Our understanding of the role of dietary macronutrients in the etiology of insulin resistance is currently limited by a paucity of credible intervention studies in humans. In contemplating such studies there are many issues that need consideration from actual study design (e.g., duration of intervention, study population, cross-over or not, nutrient formulation) to practical issues such as palatability and compliance (i.e., that terribly important issue of achievability because realistically individuals must be "free range" in order to complete studies of sufficient duration). Initiatives to support well-designed multicenter studies on diet and insulin resistance would have a major impact on our ability to treat, but more importantly to prevent, the metabolic syndrome diseases. PMID- 10812928 TI - Role of dietary factors: micronutrients. PMID- 10812929 TI - Obesity as a factor. PMID- 10812930 TI - Relationships with physical activity. AB - The evidence for beneficial effects of exercise training in the prevention and management of insulin resistance is convincing, although the mechanism remains to be fully elucidated. The increase in insulin sensitivity after a bout of exercise appears to be enhanced after training, but disappears within days of inactivity, indicating the need for regular exercise. The dose-response relationship between physical activity and insulin sensitivity deserves further study, although currently available data suggest that increases in insulin sensitivity can be achieved with regular exercise bouts of a wide range of intensities and durations. There is no evidence of a threshold for the amount of exercise that has to be performed. Extreme acute exercise that leads to muscle damage affects insulin sensitivity negatively. Both aerobic and resistance exercise training programs have been shown to be effective in increasing insulin sensitivity, and training programs that combine the two aspects may be most advantageous because they combine different mechanisms of action. PMID- 10812931 TI - Effect of aging. PMID- 10812932 TI - Genetic and environmental contributions. AB - There is a remarkable variability in insulin action in humans. Depending upon the definition of the insulin resistance syndrome, different inheritability/environmental influences on insulin action are reported. The environmental contributions to insulin resistance appear to account for approximately 50% of this syndrome. Obese and sedentary insulin-resistant individuals can see dramatic improvement in insulin sensitivity with weight reduction and fitness training. The degree to which obesity is determined by genetic influences will have a substantial impact on insulin resistance in the Western populations. Familial components also appear to account for approximately 50% of the variation and insulin action (as commonly defined by glucose metabolic effects). PMID- 10812933 TI - Stability studies of aspirin-magaldrate double layer tablets. AB - Accelerated stability testing was performed on aspirin-magaldrate double layer tablets as well as aspirin-maalox marketed double layer tablets (Ascriptin) in order to evaluate the effect of the presence of the alkaline moieties of the antacid (magaldrate and maalox) on the chemical stability of aspirin. The results were compared simultaneously with that obtained from the marketed Aspro plain tablets. The results revealed that the presence of the alkaline moieties in the tested tablets has increased the rate of aspirin decomposition and reduced its shelf-life. This effect was more pronounced for aspirin tablets containing magaldrate antacid. Determination of shelf-lives at 25 degrees C for the prepared and the marketed tablets was carried out using Arrhenius plots and the results showed that they were 35, 34.5 and 13.5 months for Aspro, Ascriptin and aspirin magaldrate double layer tablets, respectively. The effect of storage for 50 days and at different temperatures, on the crushing strength and the disintegration time of the prepared and the marked tablets showed a slight decrease in the disintegration time and the crushing strength of the tablets as the storage temperature increased. Aspro tablets did not produce the same results. The in vitro release data of the prepared aspirin-magaldrate double layer tablets and the marketed Ascriptin tablets stored for 50 days and at different storage temperatures as well as Aspro tablets stored at 70 degrees C were best fitted to the first-order kinetics model. The release data of Aspro tablets stored at 50 and 60 degrees C for 50 days were best fitted to Higuchi's model. PMID- 10812934 TI - Spectrophotometric determination of aluminium in pharmaceutical preparations by azo dyes of 1,2,4-triazole series. AB - Azodyes, derivatives of 1,2,4-triazole and pyrocatechine: 3-(3',4' dihydroxyphenylazo-1')-1,2,4-triazole (TRIAP) and 3-(3',4'-dihydroxyphenylazo-1') 5-mercapto-1,2,4-triazole (METRIAP), were used for spectrophotometric determination of Al. in composite pharmaceutical preparations. In aqueous methanolic solution at pH 6.20-6.50 Al ions form stabile, orange chelates. Molar ratio L:Al. is 2:1 (TRIAP) or 3:1 (METRIAP) and stability constants, expressed by log K, are 12,604 (METRIAP) and 8,440 (TRIAP). Other components of these preparations, particularly Mg, do not disturb Al determination. The determination results were statistically calculated and compared with those obtained by the method of atomic absorption spectrophotometry (AAS). The advantage of the developed method is easy synthesis of reagents, simple analytical procedure, stability of formed complexes, good reproducibility and accuracy of results. Using TRIAP or METRIAP the elaborated spectrophotometric method is more accurate than AAS method, for defined purpose. PMID- 10812935 TI - Improvement of water solubility and in vitro dissolution rate of gliclazide by complexation with beta-cyclodextrin. AB - Inclusion complexes of gliclazide with beta-cyclodextrin were prepared using different two methods: neutralization and recrysstalization. Host-guest interactions were studied in the solid state by X-ray diffractometry and infrared spectroscopy. The stability constant between gliclazide and beta-cyclodextrin was calculated from the phase solubility diagram. It was found that the neutralization technique and a solid complex of gliclazide with beta-cyclodextrin in a molar ratio of 1.5:1 could be used to prepare the amorphous state of drug inclusion complexes. The dissolution rates of gliclazide from the inclusion complex made by neutralization was much faster than the pure drug, physical mixture of drug and cyclodextrin, recyristalization system and also comparable to the data reported in literature. Results of this report indicate that beta cyclodextrin could be useful for the solid gliclazide formulations as it may results in a more rapid and uniform release of the drug. PMID- 10812936 TI - Spectrofluorimetric analysis of certain 4-quinolone in pharmaceuticals and biological fluids. AB - A highly sensitive spectrofluorimetric procedure is developed for the analysis of certain 4-quinolone antibiotics: sparfloxacin (I), oxolonic acid (II), flumequine (III) and enrofloxacin (IV) in their pharmaceutical dosage forms or in biological fluids. This procedure is based upon the intrinsic fluorescence in acetonitrile for sparfloxacin or upon the highly enhanced fluorescence obtained by the interaction of the drugs with AlCl3. The optimum pH for the maximum fluorescence intensity is 8-8.5 for I, 5-6 for II, III and pH 3.5 for IV. The different experimental parameters that affect the fluorescence intensity were carefully studied and incorporated into the procedure. PMID- 10812937 TI - Content of CYP3A4 inhibitors, naringin, naringenin and bergapten in grapefruit and grapefruit juice products. AB - The flavonoids, naringin and naringenin and the furanocoumarin, bergapten (5 methoxypsoralen), were detected in some fresh grapefruit and commercial grapefruit juices but were not detected in other fruit juices tested (orange; orange with apple base; dark grape; orange and mango with apple base; orange, peach, passion fruit juice). The contents of these three grapefruit constituents in commercial juice and fresh grapefruit varied from brand to brand and also from lot to lot. Juice was prepared from the fresh fruit via different methods (by hand, squeezer or blender). The naringin content, after hand-squeeze, ranged from 115 to 384 mg/l. With hand-squeeze juice production, bergapten was not detected (less than 0.5 mg/l) in two varieties of grapefruit, and naringenin was usually not in detectable levels (less than 2 mg/l) in three varieties. All three constituents were present in New Zealand grapefruit preparations (including juice by hand-squeeze) and different lots showed variation in content (1.5-, 2.3- and 4.7-fold for naringin, naringenin and bergapten, respectively). Differences in the concentrations of these three constituents, which have potential for drug interaction, may contribute to the variability in pharmacokinetics of CYP3A4 drugs and some contradictory results of drug interaction studies with grapefruit juice. PMID- 10812938 TI - Determination of the alkaloid content in different parts of some Mahonia plants by HPCE. AB - The contents of three quaternary alkaloids (berberine, palmatine, jatrorrhizine) in different parts of the genus Mahonia were determined by high-performance capillary electrophoresis (HPCE). The background electrolyte system composed of 0.1 M phosphate buffer (pH 7.0)-methanol (2:1 V/V) was found to be the most suitable solution for this separation. Brucine was used as internal standard. The linear calibration ranges were 0.004986-0.4986 mg ml-1 (r = 0.9990, n = 5) for berberine, 0.005049-0.5049 mg ml-1 (r = 0.9996, n = 5) for palmatine, and 0.005058-0.5058 mg ml-1 (r = 0.9984, n = 5) for jatrorrhizine. The relative standard deviations were 1.56%, 1.02%, and 1.60% for berberine, palmatine, and jatrorrhizine (n = 6), respectively. The recoveries were determined to be 96.00 101.66% for berberine, 100.15-102.97% for palmatine, and 96.68-102.44% for jatrorrhizine. By using proposed HPCE method, three alkaloids were well-separated within only 5.0 min. PMID- 10812939 TI - Cytotoxic activities of mono and bis Mannich bases derived from acetophenone against Renca and Jurkat cells. AB - Mannich bases of acetophenones have been disclosed to have antitumour and cytotoxic activities. 1-Phenyl-3-dimethylaminopropan-1-one hydrochloride, 1, and related piperidino, 2, and morpholino, 3, derivatives, and compound 4, which is a quaternary form of 1, were synthesized as mono Mannich bases derived from acetophenone. They were converted to corresponding bis Mannich bases, 5-8, to see whether it increases the bioactivity. The biological activity of the compounds was examined by cytotoxicity against mouse renal carcinoma (Renca) and transformed human T-lymphocyte (Jurkat) cell lines. Conversion of mono Mannich bases to corresponding bis Mannich bases remarkably increased the cytotoxicity in most cases. Quaternization procedure also improved the bioactivity in mono derivatives against Jurkat cells. Bis mannich bases 5-7 were found to be more active than 5-fluorouracil (6-23 fold) and melphalan (1.25-5 fold) against Renca cells. Except 2 and 8, the compounds synthesised were found to be more active than 5-fluorouracil (1.2-33 fold) against Jurkat cells. PMID- 10812940 TI - Long-circulating liposomes of indomethacin in arthritic rats--a biodisposition study. AB - To improve the targeting efficiency of liposomes of indomethacin to the arthritic joints, circulation half-life of the liposomes was increased by grafting amphipathic polyethylene glycol-2000 to the bilayer surface. A comparative biodistribution study was performed between the conventional liposomes (PC:CH:PE- 1:0.5:0.16) and long-circulating liposomes (PC:CH:PE-PEG--1:0.5:0.16) in arthritic rats. Pharmacokinetics of the drug changed significantly when administered in liposomal form. Pharmacokinetic parameters of the drug such as AUC0-t (trapezoidal), clearance and t1/2 (elimination half-life) changed significantly (p < 0.05) when encapsulated in liposomes. Significant difference in pharmacokinetics was observed in AUC0-t and clearance between the conventional liposomes and long-circulating liposomes. The increased AUC0-t and reduced clearance of the drug with long-circulating liposomes, increased the availability of the drug by reducing RES uptake, in turn localization in arthritic paw tissue was also increased. A concentration of 0.33 microgram of indomethacin/g of the tissue was achieved with S-liposomes after 24 h whereas it was only 0.26 microgram of drug/g of the tissue with conventional liposomes. From the study, in may be concluded that the targeting efficiency of the long-circulating liposomes was about four times more than the conventional liposomes. PMID- 10812941 TI - Lipophilicity evaluation by RP-HPLC of two homologous series of methotrexate derivatives. AB - A correlation between the structure and lipophilicity has been carried out within the homologous members of two series of aliphatic bis(amides) and lipoamino acid conjugates of the anticancer drug methotrexate. Basing on their reversed-phase HPLC behaviour, the values of the experimental parameter RQ, obtained by using different mobile phases with increasing concentration of acetonitrile, were considered as a parameter of lipophilicity of the tested compounds. A comparison with their calculated log P was also performed. PMID- 10812942 TI - Effects of sodium glycocholate and protease inhibitors on permeability of TRH and insulin across rabbit trachea. AB - The permeabilities of thyrotropin-releasing hormone (TRH) and insulin as model peptides were examined to characterize the tracheal epithelial barrier in in vitro experiments using excised rabbit trachea. TRH was not metabolized during 150 min duration of tracheal permeation and the apparent permeability coefficient (Papp) for TRH was about 3 x 10(-7) cm/s. The tracheal permeability of TRH was increased about three times by 10 mM glycocholate as a permeation enhancer. Insulin showed a slight degradation during 150 min duration of tracheal permeation, the Papp for insulin was 7 x 10(-9) cm/s. The tracheal permeability of insulin was significantly increased by 10 mM glycocholate, 1 mM bestatin (aminopeptidase B and leucine aminopeptidase inhibitor), and 10,000 KIU/ml aprotinin (trypsin and chymotrypsin inhibitor). The peptidase activities of rabbit tracheal epithelium were found to be the following; di-peptidyl aminopeptidase IV (DPP IV) > Leu-aminopeptidase > cathepsin-B > trypsin. These activities were significantly lower than those of jejunal mucosal tissues. These results suggest that the tracheal absorption of peptide drugs through the respiratory tract may contribute to the systemic delivery of these drugs following the pulmonary administration of these drugs by intratracheal insufflation and instillation. PMID- 10812943 TI - Pharmacological receptors: a century of discovery--and more. AB - A brief survey of the history of the development of the concept of the pharmacological receptor is presented. From the pioneering concepts of Paul Ehrlich, John Langley and others, receptors are described in terms of their recognition properties, their structures, transducing abilities and the impact of genomics and their role in contributing to genetic diseases. The receptor concept has firmly underpinned our advances in drug development and molecular medicine of the latter half of this century and it is clear that it will continue to drive pharmaceutical developments in the 21st century. PMID- 10812944 TI - Cholinergic receptors and neurodegenerative diseases. PMID- 10812945 TI - Nicotinic systems in central nervous systems disease: degenerative disorders and beyond. AB - Advances in the understanding of the structure, function, and distribution of central nervous system (CNS) nicotinic receptors has provided the impetus for new studies examining the role(s) that these receptors and associated processes may play in CNS functions. Further motivation has come from the realization that such receptors are changed in degenerative neurologic diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). Ongoing investigations of the molecular substructure of CNS nicotinic receptors and their pharmacology have begun to open up new possibilities for novel CNS therapeutics with nicotinic agents. Exploiting these possibilities will require understanding of the role(s) that these receptor systems play in human cognitive, behavioral, motor, and sensory functioning. Clues from careful studies of human cognition and behavior are beginning to emerge and will provide direction for studies of potentially therapeutic novel nicotinic agents. Modulation of these receptors with the ultimate goal of producing therapeutic benefits is the goal of these investigations and drug development. This paper will review studies from our laboratory and others that point to the importance of CNS nicotinic mechanisms in normal human cognitive and behavioral functioning as well as their role in disease states. In addition, this paper will examine potential clinical applications of nicotine and/or nicotinic agonists in a variety of CNS disorders with particular emphasis on structural brain disease including: movement disorders such as Parkinson's disease and Tourette's syndrome, cognitive/behavioral disorders such as Alzheimer's disease, attention deficit/hyperactivity disorder, and schizophrenia, and other more speculative applications. Important results from early therapeutic studies of nicotine and/or nicotinic agonists in these disease states are presented. For example, recent studies with nicotine and novel nicotinic agonists such as ABT-418 by our group in AD patients suggest that nicotinic stimulation can improve the acquisition and retention of verbal information and decrease errors. Preliminary results from a series of studies examining the acute and subchronic quantitative effects of nicotine on cognitive and motor functioning in Parkinson's disease suggest that acute nicotine administration and stimulation improves some aspects of cognitive and motor performance and may improve the processing speed of more complex tasks. The most likely near-term applications of novel nicotinic agonists in CNS disorders are likely to be in those disorders that are degenerative in nature, e.g. Parkinson's disease and Alzheimer's disease, or other movement disorders such as Tourette's syndrome. The most likely direct therapeutic role for nicotinic agonists is as augmentation therapy in combination with other agents rather than as monotherapy, except early in disease states or as a prophylactic or preventative treatment. PMID- 10812946 TI - Central nicotinic receptor ligands and pharmacophores. AB - Multiple populations of pentameric nicotinic acetylcholinergic (nACh) receptors exist and several may be classified as central or neuronal. Neuronal nACh receptors, however, are primarily of the alpha 4 beta 2 and alpha 7 types, and these have been the focus of most recent investigations aimed at the development of novel agents and identification of pharmacophores. Selectivity data are limited. Furthermore, because several populations of nACh receptors might indirectly influence a given functional effect, it is difficult to discuss structure-activity relationships (SAR) in terms of differential SAR, or to formulate SAR on the basis of functional studies. For the most part, studies are limited to the formulation of structure-affinity relationships (SAFIR) for the binding of agents at nACh receptors, and for these the alpha 4 beta 2 population has been the most extensively investigated. SAFIR and newer agents are reviewed here with reference to earlier studies. Novel agents now have been identified that bind with up to 30 times higher affinity than nicotine and these are providing new insight into the understanding of nACh receptors. PMID- 10812947 TI - Structural aspects of high affinity ligands for the alpha 4 beta 2 neuronal nicotinic receptor. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are a heterogeneous family of related ion channels that are widely distributed throughout the central and peripheral nervous systems. They all share a common architecture of five subunit proteins that combine at the cell surface to create a ligand-gated cation permeable pore. Significant effort is currently being expended by medicinal chemistry teams to synthesize ligands that exhibit selectivity for central over peripheral nAChR subtypes. Within the CNS, multiple nAChR subtypes are recognized, and the discovery of ligands exhibiting selectivity among these subtypes offers an opportunity for the development of novel therapeutic agents. The alpha 4 beta 2 subtype is one of the most abundant nAChR subtypes within the CNS, and has been the primary focus of high affinity ligand design. Nicotine (1), and more recently, epibatidine (2) have served as structural templates for the design of the majority of active compounds. Although the diversity of nAChR ligands is growing, the structural requirements necessary for high affinity binding with the alpha 4 beta 2 receptor remain poorly understood. The putative pharmacophoric elements common to all potent alpha 4 beta 2 ligands include (1) a basic or quaternized nitrogen atom, and (2) a less basic nitrogen or a carbonyl oxygen that presumably interact with electron rich and electron deficient sites on the receptor, respectively. The family of currently known high affinity analogs consists of a diverse array of azacycles containing a basic amine. Several additional basic amine fragments have been identified, including the pyrrolizidine nucleus (exemplified by 8) and the 2-azabicyclo[2.2.1]heptane skeleton (exemplified by 9). In addition, we have found that the furo[2,3 b]pyridine heterocycle (compound 10) serves as useful bioisosteric replacement for the pyridyl substituent of nicotine. A preliminary pharmacophore model is proposed in which a reasonable superposition of the putative pharmacophoric elements of the diverse array of high affinity ligands for the alpha 4 beta 2 nAChR reported herein may be accommodated. PMID- 10812948 TI - Recombinant human receptors and functional assays in the discovery of altinicline (SIB-1508Y), a novel acetylcholine-gated ion channel (nAChR) agonist. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are a class of ion channels with significant potential as molecular targets for the design of drugs to treat a variety of CNS disorders. The discovery that neuronal nAChRs are further subdivided into multiple subtypes suggests that drugs which act selectively at specific nAChR subtypes might effectively treat Parkinson's disease (PD), Alzheimer's disease (AD), schizophrenia, ADHD, depression, anxiety or pain without the accompanying adverse side effects associated with non-selective agents such as nicotine (1) and epibatidine. Altinicline (SIB-1508Y) is a novel, small molecule designed to selectively activate neuronal nAChRs and is undergoing clinical evaluation for the treatment of PD. It was selected from a series of compounds primarily on the basis of results from functional assays, including (a) measurement of Ca2+ flux in stable cell lines expressing specific recombinant human neuronal nAChR subtypes; (b) determination of in vitro and in vivo neurotransmitter release; (c) in vivo models of PD. Biological data on both altinicline and the series of compounds from which it was selected are reported. PMID- 10812949 TI - Receptors in neurodegenerative diseases, muscarinic cholinergic receptors. PMID- 10812950 TI - Design and development of selective muscarinic agonists for the treatment of Alzheimer's disease: characterization of tetrahydropyrimidine derivatives and development of new approaches for improved affinity and selectivity for M1 receptors. AB - Cholinergic neurons degenerate in Alzheimer's disease, resulting in cognitive impairments and memory deficits, and drug development efforts have focused on selective M1 muscarinic agonists. 5-(3-Ethyl-1,2,4- oxadiazol-5-yl)-1,4,5,6 tetrahydropyrimidine trifluoroacetic acid (CDD-0102) stimulates M1 muscarinic receptors in rat brain [Messer, W.S., Jr., Abuh, Y.F., Liu, Y., Periyasamy, S., Ngur, D.O., Edgar, M.A., El-Assadi, A.A., Sbeih, S., Dunbar, P.G., Roknich, S., Rho, T., Fang, Z., Ojo, B., Zhang, H., Huzl, J.J., III, Nagy, P.I., 1997a. J. Med. Chem. 40, 1230-1246.] and improves memory function in rats with lesions of the basal forebrain cholinergic system. Moreover, CDD-0102 exhibits oral bioavailability, few side effects and low toxicity, and thus represents a viable candidate for clinical studies. Despite the development of functionally selective agonists such as xanomeline and CDD-0102, there is room for improvements in ligand affinity and selectivity. The high degree of amino acid homology within transmembrane domains has hindered the development of truly selective agonists. Site-directed mutagenesis, biochemical and molecular modeling studies have identified key amino acid residues such as Thr192 and Asn382 in the binding of agonist to M1 receptors [Huang, X.P., Nagy, P.I., Williams, F.E., Peseckis, S.M., Messer, W.S., Jr., 1999. Br. J. Pharmacol. 126, 735-745.]. Recent work has implicated residues at the top of transmembrane domain VI in the binding of muscarinic agonists and activation of M1 receptors [Huang, X.P., Williams, F.E., Peseckis, S.M., Messer, W.S., Jr., 1998. J. Pharmacol. Exp. Ther. 286, 1129 1139.]. Thus, residues such as Ser388 represent molecular targets for the further development of agonists with improved M1 receptor affinity, selectivity and activity. PMID- 10812951 TI - CI-1017, a functionally M1-selective muscarinic agonist: design, synthesis, and preclinical pharmacology. AB - The five muscarinic receptor subtypes (M1-M5) are characterized by seven helices that define a transmembrane cavity which serves as the binding pocket for agonists and antagonists. The five cavities appear to be topographically different enough to permit subtype selectivity among antagonists but not among classical agonists which tend to be smaller in size than antagonists. It was reasoned that synthesis of muscarinic agonists longer/larger than their classical counterparts might result in subtype selectivity. M1 subtype selectivity was found in a class of 1-azabicyclo[2.2.1]heptan-3-one, O-(3-aryl-2-propynyl) oximes. One of these, CI-1017, improved spatial memory of hippocampally deficient mice and nbM-lesioned rats at doses of 1.0-3.2 and 0.1-0.3 mg/kg, respectively, while producing parasympathetic side effects only at very high doses (100-178 mg/kg). Additionally, CI-1017 inhibited production of amyloidogenic A beta and increased secretion of soluble APP. Thus, CI-1017, besides treating AD symptomatically, may also retard its progression. CI-1017 has recently completed phase I clinical trials. PMID- 10812952 TI - Ligands for the common allosteric site of acetylcholine M2-receptors: development and application. AB - Ligands for the allosteric site of acetylcholine M2 receptors are able to retard the dissociation of simultaneously bound ligands for the orthosteric site. This effect promotes receptor occupation by the orthosteric ligand. The allosteric effect opens various therapeutic perspectives, e.g., in organophosphorus poisoning. The aim of our studies was to optimize the affinity of the modulators for the common allosteric binding site of muscarinic M2 receptors, the orthosteric site of which was liganded with the N-methylscolopamine. The phthalimido substituted hexane-bisammonium compound W84 served as a starting point. Previous molecular modelling studies revealed two positive charges and two aromatic imides in a sandwich-like arrangement to be essential for a high allosteric potency. A three-dimensional quantitative structure activity relationship (3D QSAR) analysis predicted compounds with substituents of increasing size on the lateral imide moieties to enhance the affinity for the allosteric binding site. Thus, we synthesized and pharmacologically evaluated compounds bearing "saturated" phthalimide moieties as well as phthalimidines with substituents of systematically increasing size in position 3 or on the aromatic ring at one or both ends of the molecule. Within each series, QSAR could be derived: 1. "Saturation" of the aromatic ring of the phthalimide moiety results in less potent compounds. 2. Increasing the size of the substituents in position 3 of the phthalimide enhances the potency. 3. Putting substituents on the aromatic part of the phthalimide increases the potency more effectively: the introduction of a methyl group in position 5 gave a compound with a potency in the nanomolar concentration range which was subsequently developed as the first radioligand for the allosteric binding site. PMID- 10812953 TI - Receptors in cardiovascular disease: review and introduction. AB - Despite recent encouraging declines, cardiovascular disease (CVD) is still responsible for about 50% of premature death in the Western industrialized countries, greater than cancer, AIDS and accidents, combined. Different aspects of the disease have been considered and the main currently available and possible future drugs whose effect is based on interaction with a receptor have been reviewed. Catecholamines receptors ligands, mainly beta-blockers, and the new angiotensin II antagonists represent the most important classes among the established therapies. Investigational approaches such as the oral glycoprotein GPIIb/IIIa antagonists and endothelin, adenosine and neuropeptide Y receptors ligands are discussed. Receptorology represents just a part of the therapeutical approach to CVD, where other classes of drugs with enzyme or ionic channel based mechanisms are largely used and innovative therapies based on the most advanced research techniques could early become reality. PMID- 10812954 TI - Adrenoceptor subclassification: an approach to improved cardiovascular therapeutics. AB - The subdivision of alpha adrenoceptors into the alpha 1 and alpha 2 classes was the impetus for the design of the selective alpha 1-adrenoceptor antagonists, which remain useful antihypertensives. alpha 2-Adrenoceptor agonists also have application as antihypertensive drugs, based on their ability to reduce sympathetic outflow. Likewise, subdivision of the beta adrenoceptors has lead to the development of selective beta 1-adrenoceptor antagonists as antihypertensive and selective beta 2 agonists as bronchodilators. In the past decade, both the alpha 1 and alpha 2 adrenoceptors have been further subdivided, each into three subclasses. In addition, there is strong functional evidence to suggest the presence of additional adrenoceptor subtypes, such as the "alpha 1L" adrenoceptor and "beta 4" adrenoceptor. alpha 1A (or alpha 1L)-Adrenoceptor antagonists have been evaluated for benign prostatic hyperplasia (BPH), and selective alpha 1A agonists for stress incontinence. Gene knockout experiments in mice suggest an important role for the alpha 1B adrenoceptor in the control of vascular tone. Hence, selective alpha 1B antagonists may offer a new approach toward hypertension. Although targeting of specific adrenoceptors can be used to optimize the therapeutic profile of a drug, there are also cases where blockade of multiple adrenoceptors is desirable, as with the alpha/beta-adrenoceptor antagonist carvedilol in congestive heart failure. It is possible that combination of affinities for selected adrenoceptor subtypes within a single molecule may be desirable for certain applications. PMID- 10812955 TI - The alpha 1a and alpha 1b-adrenergic receptor subtypes: molecular mechanisms of receptor activation and of drug action. AB - In this chapter we summarize some aspects of the structure-functional relationship of the alpha 1a and alpha 1b-adrenergic receptor subtypes related to the receptor activation process as well as the effect of different alpha-blockers on the constitutive activity of the receptor. Molecular modeling of the alpha 1a and alpha 1b-adrenergic receptor subtypes and computational simulation of receptor dynamics were useful to interpret the experimental findings derived from site directed mutagenesis studies. PMID- 10812956 TI - Alpha 1-adrenoreceptor antagonists bearing a quinazoline or a benzodioxane moiety. PMID- 10812957 TI - Selection, design and evaluation of new radioligands for PET studies of cardiac adrenoceptors. AB - Changes in the numbers of human cardiac adrenoceptors (ARs) are associated with various diseases, such as myocardial ischemia, congestive heart failure, cardiomyopathy and hypertension. There is a clear need for capability to assess human cardiac ARs directly in vivo. Positron emission tomography (PET) is an imaging technique that provides this possibility, if effective radioligands can be developed for the targeted ARs. Here, the status of myocardial AR radioligand development for PET is described. Currently, there exist effective radioligands for imaging beta-ARs in human myocardium. One of these, [11C](S)-CGP 12177, is applied extensively to clinical research with PET, sometimes with other tracers of other aspects of the noradrenalin system. Alternative radioligands are in development for beta-ARs, including beta 1-selective radioligands. A promising radioligand for imaging myocardial alpha 1-ARs, [11C]GB67, is now being evaluated in human PET experiments. PMID- 10812958 TI - Enigmatic receptors. AB - The identification of new binding sites raises the problem of defining their role, if any. At times they are shown to be pharmacological receptors, in a strict sense, as they fulfill certain requirements, and a precise physiological role and function, and an endogenous ligand (neurotransmitter) are discovered. At other times, however, neither a clear physiological role nor an endogenous ligand are found, but the term "receptor" is still used, although it may not be a proper one in the conventional pharmacological sense. Furthermore, no clear intracellular signalling transduction pathway is defined and, as a consequence, it is not possible to determine whether drugs binding to these receptors act as agonists or antagonists. What their structure and biological function are and how they mediate the pharmacological effects of ligands may remain for a long time an enigma. The matter, in any case, is of great interest to researchers of different areas, especially to medicinal chemists who foresee novel potential targets for therapeutic interventions. In this meeting one section is dedicated to two examples of this kind of receptors: imidazoline (I) and sigma (sigma) receptors. PMID- 10812959 TI - Imidazoline receptors: a challenge. AB - The hypotensive effect of imidazoline-like drugs (IMs) directly injected into the rostroventrolateral part of the brainstem (NRL/RVLM) was shown to involve non adrenergic imidazoline specific receptors (IRs). Some IMs caused hypotension when injected there, irrespective of their affinity and selectivity for any alpha adrenoceptor subtype. Compounds, such as LNP 509, S 23515, S 23757 or benazoline with very high selectivities for IRs over alpha 2-adrenoceptors (A2Rs), became available recently. Some of these compounds (LNP 509, S 23515) caused hypotension when injected alone into the NRL/RVLM region. Nevertheless, high selectivity for IRs will not predict by its own the capability of IMs to elicit hypotension as some of these substances behaved as antagonists towards the hypotensive effects of the latter. As far as hybrid drugs, i.e., with mixed binding profiles (I1/alpha 2), were concerned, a significant correlation has been reported between their central hypotensive effect and their affinity for IRs. Imidazoline antagonists, such as idazoxan, were repeatedly shown to competitively prevent and reverse the centrally induced hypotensive effect of IMs. The sole stimulation of A2Rs within the NRL/RVLM region was not sufficient to decrease blood pressure as much as IMs did, as shown by the lack of significant blood pressure lowering effect of alpha-methylnoradrenaline (alpha-MNA). No correlation was observed between affinity of IMs for A2Rs and their central hypotensive effects. It is also noticeable that yohimbine, an A2Rs antagonist, was repeatedly shown to abolish the hypotensive effect of hybrids but usually in a non-competitive manner. Mutation of A2Rs was shown to prevent the hypotensive effects of centrally acting drugs. It is concluded that (i) drugs highly selective for I1Rs over A2Rs can reduce blood pressure by their own; (ii) the central hypotensive effect of IMs needs implication of IRs and appears to be facilitated by additional activation of A2Rs; and (iii) this effect requires intact A2Rs along the sympathetic pathways. PMID- 10812960 TI - Sigma receptors: recent advances and new clinical potentials. AB - Several recent advances are leading to a better understanding of sigma receptors. Here we focus on our recent findings regarding cellular functions of sigma-2 receptors and discuss their possible clinical implications. Agonists at sigma-2 receptors induced changes in cell morphology and apoptosis in various cell types. Sigma-2 receptor activation produced both transient and sustained increases in [Ca++]i, derived from different intracellular stores. These changes in [Ca++]i and cytotoxic effects are mediated by intracellular sigma-2 receptors. Sigma-2 agonists induced apoptosis in drug-resistant cancer cells, enhanced the potency of DNA damaging agents, and down-regulated expression of p-glycoprotein mRNA. Thus, sigma-2 receptor agonists may be useful in treatment of drug-resistant cancers. Sigma radioligands have been used in tumor imaging. We also discuss how sigma-2 antagonists might prevent the irreversible motor side effects of typical neuroleptics. Sigma-2 receptors may subserve a novel signalling pathway to apoptosis, involved in regulation of cell proliferation and/or viability. PMID- 10812961 TI - Excitatory amino acid receptors. PMID- 10812962 TI - Excitatory amino acid agonists and antagonists: pharmacology and therapeutic applications. AB - Glutamic acid is the major excitatory neurotransmitter in the mammalian central nervous system (CNS). Specific receptors bind glutamate and some of these when activated open an integral ion channel and are thus known as ionotropic receptors. Within the ionotropic family of glutamate receptors, three major subtypes have been identified using classical specific agonist activation, selective competitive antagonists together with their structural heterogeneity. These receptors have thus been named N-methyl-D-aspartate (NMDA), alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate receptors. The NMDA receptor has sites in addition to its agonist-binding site and these seem to either positively or negatively modulate the agonist effect. The NMDA receptor also is unique in that another amino acid, glycine, acts as a co-agonist with glutamate. Changes in glutamate transmission have been associated with a number of CNS pathologies; these include, acute stroke, chronic neurodegeneration, chronic pain, depression, drug dependency, epilepsy, Parkinson's Disease and schizophrenia. PMID- 10812963 TI - Metabotropic glutamate receptors: a structural view point. PMID- 10812964 TI - Synthesis and pharmacological properties of novel glycine antagonists. AB - The NMDA receptor is an ionotropic receptor complex widely distributed in the central nervous system and its activation, particularly in hypoxic conditions such as stroke, traumatic head injury and hypoglycemia, results in a massive influx of calcium ions into the post-synaptic neurones, leading to cell death through the activation of several neurotoxic cascades. The NMDA receptor is a unique ionotropic receptor complex because its activation requires the simultaneous binding of glutamate and glycine and selective antagonists at the glycine binding site are endowed with a better side-effect profile than competitive NMDA antagonists. Then, considerable efforts have been devoted to find potent and selective ligands, resulting in the identification of several classes of glycine antagonists. The research at Glaxo Wellcome has been aimed at the identification of novel in vivo active glycine antagonists, and led to the synthesis and pharmacological characterization of a number of novel, potent and systemically active compounds belonging to different chemical classes. PMID- 10812965 TI - Receptors in neurodegenerative diseases. AB - The ability of trophic factors to regulate developmental neuronal survival and adult nervous system plasticity suggests the use of these molecules to treat neurodegeneration associated with human diseases, such as Alzheimer's, Huntington's and Parkinson's disease, of amyotrophic lateral sclerosis and peripheral sensory neuropathies. Recent biological data on the neutrotrophins NGF and BDNF, on GDNF, CNTF and IGF-I are discussed together with first results from clinical trials. Literature is presented on the three-dimensional structures of these trophic factors and on models proposed for ligand-receptor interactions. Substantial progress has been made in the understanding of the mechanisms of apoptosis. The cascade consisting of interaction of apoptosis-inducing ligands with death receptors, the coupling of this complex to adaptor proteins via death domains, the further recruitment of procaspases via death effector or caspase recruitment domains and the execution of cell death via the effector caspases is briefly outlined. PMID- 10812966 TI - Neurotrophin receptor structure and interactions. AB - Although ligand-induced dimerization or oligomerization of receptors is a well established mechanism of growth factor signaling, increasing evidence indicates that biological responses are often mediated by receptor trans-signaling mechanisms involving two or more receptor systems. These include G protein coupled receptors, cytokine, growth factor and trophic factor receptors. Greater flexibility is provided when different signaling pathways are merged through multiple receptor signaling systems. Trophic factors exemplified by NGF and its family members, ciliary neurotrophic factor (CNTF) and glial derived neurotrophic factor (GDNF) all utilize increased tyrosine phosphorylation of cellular substrates to mediate neuronal cell survival. Actions of the NGF family of neurotrophins are not only dictated by ras activation through the Trk family of receptor tyrosine kinases, but also a survival pathway defined by phosphatidylinositol-3-kinase activity (Yao and Cooper, 1995), which gives rise to phosphoinositide intermediates that activate the serine/threonine kinase Akt/PKB (Dudek et al., 1997). Induction of the serine-threonine kinase activity is critical for cell survival, as well as cell proliferation. Hence, for many trophic factors, multiple proteins constitute a functional multisubunit receptor complex that activates ras-dependent and ras-independent intracellular signaling. The NGF receptors provide an example of bidirectional crosstalk. In the presence of TrkA receptors, p75 can participate in the formation of high affinity binding sites and enhanced neurotrophin responsiveness leading to a survival or differentiation signal. In the absence of TrkA receptors, p75 can generate, in only specific cell populations, a death signal. These activities include the induction of NF kappa B (Carter et al., 1996); the hydrolysis of sphingomyelin to ceramide (Dobrowsky et al., 1995); and the pro-apoptotic functions attributed to p75. Receptors are generally drawn and viewed as isolated integral membrane proteins which span the lipid bilayer, with signal transduction proceeding in a linear step-wise fashion. There are now numerous examples which indicate that each receptor acts not only in a linear, independent manner, but can also influence the activity of other cell surface receptors, either directly or through signaling intermediates. Which step and which intermediates are utilized for crosstalk between the receptors is a critical question. For neurotrophins, their primary function in sustaining the viability of neurons is counterbalanced by a receptor mechanism to eliminate cells by an apoptotic mechanism. It is conceivable that this bidirectional system may be utilized selectively during development and in neurodegenerative diseases. PMID- 10812967 TI - The RET receptor tyrosine kinase: activation, signalling and significance in neural development and disease. AB - The RET receptor tyrosine kinase was first identified in a screen for human oncogenes and has subsequently been linked to several human syndromes: Hirschprung's disease, multiple endocrine neoplasia types 2A and 2B and familial thyroid carcinoma. Interestingly, all of the tissues affected by mutations in RET are derived from the neural crest during development. RET transduces a signal following activation by ligands of the glial cell line-derived neurotrophic factor (GDNF) family of neurotrophins which currently comprises GDNF, neuturin (NTN), artemin (ART) and persephin (PSP). To activate RET they form a tripartite complex with RET and a member of a family of four extracellular, GPI-linked alpha receptors (GFR alpha 1-4). Specificity is achieved by each GFR alpha binding only one member of the GDNF family with high affinity. Current evidence indicates that signal transduction by RET activates several second messenger systems including the PLC gamma, Ras, JNK and inositol phosphate pathways. Targeted mutagenesis in transgenic mice has shown that Ret, GFR alpha 1 and GDNF are required for multiple developmental events including development of the enteric nervous system (ENS) affected in Hirschsprung's disease. We describe experiments in chick neural crest cells which provide evidence for the normal function of RET and the basis of the defect in Hirschsprung's disease. PMID- 10812968 TI - The ciliary neurotrophic factor and its receptor, CNTFR alpha. AB - Ciliary neurotrophic factor (CNTF) is expressed in glial cells within the central and peripheral nervous systems. CNTF stimulates gene expression, cell survival or differentiation in a variety of neuronal cell types such as sensory, sympathetic, ciliary and motor neurons. In addition, effects of CNTF on oligodendrocytes as well as denervated and intact skeletal muscle have been documented. CNTF itself lacks a classical signal peptide sequence of a secreted protein, but is thought to convey its cytoprotective effects after release from adult glial cells by some mechanism induced by injury. Interestingly, mice that are homozygous for an inactivated CNTF gene develop normally and initially thrive. Only later in adulthood do they exhibit a mild loss of motor neurons with resulting muscle weakness, leading to the suggestion that CNTF is not essential for neural development, but instead acts in response to injury or other stresses. The CNTF receptor complex is most closely related to, and shares subunits with the receptor complexes for interleukin-6 and leukemia inhibitory factor. The specificity conferring alpha subunit of the CNTF complex (CNTFR alpha), is extremely well conserved across species, and has a distribution localized predominantly to the nervous system and skeletal muscle. CNTFR alpha lacks a conventional transmembrane domain and is thought to be anchored to the cell membrane by a glycosyl-phosphatidylinositol linkage. Mice lacking CNTFR alpha die perinatally, perhaps indicating the existence of a second developmentally important CNTF-like ligand. Signal transduction by CNTF requires that it bind first to CNTFR alpha, permitting the recruitment of gp130 and LIFR beta, forming a tripartite receptor complex. CNTF-induced heterodimerization of the beta receptor subunits leads to tyrosine phosphorylation (through constitutively associated JAKs), and the activated receptor provides docking sites for SH2 containing signaling molecules, such as STAT proteins. Activated STATs dimerize and translocate to the nucleus to bind specific DNA sequences, resulting in enhanced transcription of responsive genes. The neuroprotective effects of CNTF have been demonstrated in a number of in vitro cell models as well as in vivo in mutant mouse strains which exhibit motor neuron degeneration. Intracerebral administration of CNTF and CNTF analogs has also been shown to protect striatal output neurons in rodent and primate models of Huntington's disease. Treatment of humans and animals with CNTF is also known to induce weight loss characterized by a preferential loss of body fat. When administered systemically, CNTF activates downstream signaling molecules such as STAT-3 in areas of the hypothalamus which regulate food intake. In addition to its neuronal actions, CNTF and analogs have been shown to act on non-neuronal cells such as glia, hepatocytes, skeletal muscle, embryonic stem cells and bone marrow stromal cells. PMID- 10812969 TI - Rediscovering good old friend IGF-I in the new millenium: possible usefulness in Alzheimer's disease and stroke. AB - Much research has been done over the past two decades on the role of insulin-like growth factors I and II (IGF) in the maintenance of normal body homeostasis, especially in regard to various endocrine functions, growth and aging. For example, IGF-I is a well established promoter of tissue growth and has been used in the clinics for the treatment of growth related disorders, even being abused by athletes to enhance performance in competitions. In contrast, comparatively limited attention has been given to the potential significance of the IGFs in the central nervous system. Over the past few years, we have studied the trophic as well as neuromodulatory roles of the IGFs in the brain. IGF-I and IGF-II are potent modulators of acetylcholine release, IGF-I inhibiting release while IGF-II is a potent stimulant. Moreover, only the internalization of the IGF-I receptor complex was blocked by an inhibitor of phosphotyrosylation. This is in accordance with the differential nature of the IGF-I and IGF-II receptors, the former being a tyrosine kinase receptor while the later is a single transmembrane domain protein bearing binding sites for 6-mannose phosphate containing residues. The activation of IGF-I receptors protected neurons against cell death induced by amyloidogenic derivatives likely by an intracellular mechanism distinct from those involved in the regulation of acetylcholine release and neuronal growth. The stimulation of IGF-I receptors can activate intracellular pathways implicating a PI3/Akt kinase and CREB phosphorylation or modulate the production of free radicals. The effects, particularly those of IGF-I on key markers of the Alzheimer's (AD) brains namely cholinergic dysfunction, neuronal amyloid toxicity, tau phosphorylation and glucose metabolism suggest the potential usefulness of this growth factor in the treatment of neurodegenerative diseases. However, the poor bioavailability, enzymatic stability and brain penetration of IGF-I hamper progress in this regard. The recent development of a small, non peptidyl mimetic of insulin able to directly activate the insulin receptor [Zhang, B., Salituro, G., Szalkowski, D., Li, Z., Zhang, Y., Royo, I., Vilella, D., Diez, M.T., Pelaez, F., Ruby, C., Kendall, R.L., Mao, X., Griffin, P., Calaycay, J., Zierath, J.R., Heck, J. V., Smith, R.G., Moller, D.E., 1999. Science, 284, 974-977] suggests that a similar strategy could be used for IGF-I and the IGF-I receptor leading to the characterization of IGF-I mimics of potential clinical usefulness. PMID- 10812970 TI - Apoptosis induced by death receptors. AB - Death receptors belong to the TNF receptor family and are characterised by an intracellular death domain that serves to recruit adapter proteins such as TRADD and FADD and cysteine proteases such as Caspase-8. Activation of Caspase-8 on the aggregated receptor leads to apoptosis. Triggering of death receptors is mediated through the binding of specific ligands of the TNF family, which are homotrimeric type-2 membrane proteins displaying three receptor binding sites. There are various means of modulating the activation of death receptors. The status of the ligand (membrane-bound vs. soluble) is critical in the activation of Fas and of TRAIL receptors. Cleavage of membrane-bound FasL to a soluble form (sFasL) does not affect its ability to bind to Fas but drastically decreases its cytotoxic activity. Conversely, cross-linking epitope-tagged sFasL with anti-tag antibodies to mimic membrane-bound ligand results in a 1000-fold increase in cytotoxicity. This suggests that more than three Fas molecules need to be aggregated to efficiently signal apoptosis. Death receptors can also be regulated by decoy receptors. The cytotoxic ligand TRAIL interacts with five receptors, only two of which (TRAIL-R1 and -R2) have a death domain. TRAIL-R3 is anchored to the membrane by a glycolipid and acts as a dominant negative inhibitor of TRAIL mediated apoptosis when overexpressed on TRAIL-sensitive cells. Intracellular proteins interacting with the apoptotic pathway are potential modulators of death receptors. FLIP resembles Caspase-8 in structure but lacks protease activity. It interacts with both FADD and Caspase-8 to inhibits the apoptotic signal of death receptors and, at the same time, can activate other signalling pathways such as that leading to NF-kappa B activation. PMID- 10812971 TI - Hijacked receptors. AB - Pharmacological receptors are typically defined by their selectivity of ligand recognition, including where appropriate stereoselectivity of interaction. It is increasingly clear that receptors may, in fact, be promiscuous species. This promiscuity arises at several levels of organization: two appear to be of particular importance. A given ligand-receptor complex may couple with different effectors and may generate quite different physiological responses: this is particularly common, although not uniquely so, for G protein-coupled receptors. Or a single receptor may recognize fundamentally different ligands often of significantly different characteristics: a number of viruses gain entry to cells through their interaction at receptors for neurotransmitters, peptides or hormones. PMID- 10812972 TI - ICAM-1 receptors and cold viruses. AB - Human rhinoviruses (HRVs), the single most important etiologic agent of common colds, are small viruses composed of an icosahedral protein shell that encapsidates a single, positive RNA strand. Multiplication of HRVs occurs in the cytoplasm of the host cell. To produce infection, HRVs must first attach to specific cellular receptors embedded in the plasma membrane. Ninety percent of HRVs immunogenic variants use as receptor intercellular adhesion molecule-1 (ICAM 1), a cell surface glycoprotein that promotes intercellular signaling in processes derived from inflammation response. As HRV receptor, ICAM-1 positions the virus to within striking distance of the membrane, and then triggers a conformational change in the virus that ultimately results in delivery of the viral RNA genome into the cytoplasm, across a lipid bilayer. The interaction between ICAM-1 and HRVs has been analyzed by the combination of crystal structures of HRVs and ICAM-1 fragments with electron microscopy reconstructions of the complexes. The resulting molecular models are useful to address questions about receptor recognition, binding specificity, and mechanisms by which ICAM-1 induces virus uncoating. PMID- 10812973 TI - Viral-encoded G-protein coupled receptors: new targets for drug research? PMID- 10812974 TI - Chemokine receptors: interaction with HIV-1 and viral-encoded chemokines. AB - Chemokines are a superfamily of proteins that play a central role in immune and inflammatory reactions and in viral infections. About 50 different chemokines divided in four subfamilies are known, CXC, CC, C, and CX3C. Chemokine receptors can function as entry/fusion co-receptors for human immunodeficiency virus (HIV) 1 infection, and regulation of receptor expression by cytokines may be relevant for viral infection. Posttranslational processing of chemokines can profoundly affect their interaction with receptors. The serine protease CD26/dipeptidyl peptidase IV (CD26/DPP IV) removes NH2-terminal dipeptides from several chemokines and profoundly affect their biological activity. Kaposi's sarcoma (KS) associated herpes virus 8 encodes for three chemokine-like proteins that show homology with MIP cluster of CC chemokines. These viral chemokines possess a partial agonist activity for certain chemokine receptors and may function as receptor antagonists. This biological activity could represent a strategy developed by the virus to subvert immunity impairing the generation of an effective anti-viral immune response. PMID- 10812975 TI - General topics and perspectives PMID- 10812976 TI - Pharmacological evidence of muscarinic receptor heterodimerization. PMID- 10812977 TI - Constitutive activity of G protein coupled receptors and drug action. PMID- 10812978 TI - New dimensions in G protein signalling: G beta 5 and the RGS proteins. AB - The beta gamma complex of G-proteins regulates effectors independently of the G alpha subunits, such that upon activation G proteins give may signal downstream along one or both pathways. The G beta 5 isoform exhibits much less homology with other G beta isoforms (approximately 50%) and is preferentially expressed in brain. The G beta 5 isoform exhibits novel properties in its activation of effector pathways such as MAPK, phospholipase C-beta, and adenylyl cyclase type II when compared to G beta 1. Recently specific native complexes between G beta 5 and the regulator of G protein signaling (RGS) protein-7 (RGS7) and between G beta 5L (a splice variant with a 42 amino acid N-terminal extension) and RGS9 have been isolated from different retinal fractions. Such findings are not accounted for by current models as only the G alpha subunits and not G beta had been previously implicated in RGS protein function. These recent novel observations further reinforce the view of G beta 5 as a unique and highly specialized G protein subunit. PMID- 10812979 TI - Kappa opioid agonists as targets for pharmacotherapies in cocaine abuse. AB - Kappa opioid receptors derive their name from the prototype benzomorphan, ketocyclazocine (1a) which was found to produce behavioral effects that were distinct from the behavioral effects of morphine but that were antagonized by the opioid antagonist, naltrexone. Recent evidence suggests that agonists and antagonists at kappa opioid receptors may modulate the activity of dopaminergic neurons and alter the neurochemical and behavioral effects of cocaine. Kappa agonists blocked the effects of cocaine in squirrel monkeys in studies of cocaine discrimination and scheduled-controlled responding. Studies in rhesus monkeys suggested that kappa opioids may antagonize the reinforcing effects of cocaine. These studies prompted the synthesis and evaluation of a series of kappa agonists related to the morphinan, L-cyclorphan (3a) and the benzomorphan, L-cyclazocine (2). We describe the synthesis and preliminary evaluation of a series of morphinans, structural analogs of cyclorphan 3a-c, the 10-keto morphinans 4a and b, and the 8-keto benzomorphan 1b, structurally related to ketocyclazocine (1a). In binding experiments L-cyclorphan (3a), the cyclobutyl (3b), the tetrahydrofurfuryl 3c and the 10-keto 4b analogs had high affinity for mu (mu), delta (delta) and kappa (kappa) opioid receptors. Both 3a and 3b were more selective for the kappa receptor than the mu receptor. However, 3b was 18-fold more selective for the kappa receptor in comparison to the delta receptor, while cyclorphan (3a) had only a 4-fold greater affinity for the kappa receptor in comparison to the delta receptor. The cyclobutyl compound 3b was found to have significant mu agonist properties, while 3a was a mu antagonist. All compounds were also examined in the mouse tail flick and writhing assay. Compounds 3a and 3b were kappa agonists. Correlating with the binding results, compound 3a had some delta agonist properties, while 3b was devoid of any activity at the delta receptor. In addition, compounds 3a and 3b had opposing properties at the mu opioid receptor. The cyclobutyl compound 3b was found to have significant mu agonist properties, while 3a was a mu antagonist. PMID- 10812980 TI - [Antibiotics as inhibitors of the nitrate reductase in human oral fluid]. PMID- 10812981 TI - [The effect of the immunomodulator likopid on cytokine synthesis and on the activation molecules of the peripheral blood lymphocytes in patients with lichen ruber planus]. AB - Clinical and immunological efficacy of immunomodulator likopide has been studied in patients with lichen planus. The levels of peripheral blood lymphocytes and CD3+ lymphocytes, containing antiinflammatory cytokine gamma-interferon in the cytoplasm, are increased in these patients. Likopide therapy led to a notable decrease in the content of lymphocytes and CD3+ lymphocytes containing this cytokine. The count of lymphocytes containing inflammatory cytokine interleukin-4 (IL-4) in the cytoplasm is the same in patients with lichen planus and donors. Likopide stimulated the production of this interleukin by peripheral blood mononuclears. Expression of early activation antigen CD69 on CD3+ and CD3+CD8+ lymphocytes can be a sign of improvement of these cells' function. The drug had a pronounced clinical effect in virtually all forms of lichen planus, which may be due to suppression of the activity of TH1 cells producing inflammatory cytokines and an increase in the functional activity of TH2 cells producing antiinflammatory cytokines and to normalization of the functional activity of T lymphocytes, manifesting by a higher expression of early activation antigen CD69 involved in the production of IL-2. PMID- 10812982 TI - [The analgesic action of the preparation Ketanov in the combined treatment of patients with pulpitis and periodontitis]. AB - The purpose of this clinical investigation was to compare analgetic effect of analgin and Ketanov on posttreatment pain after canal filling in 50 patients with pulpitis and periodontitis. Clinical data have shown Ketanov effectiveness, absence of side effects. Marked reduction of pain was shown in all patients getting Ketanov, no analgetic effect was revealed in 20% patients getting analgin. PMID- 10812983 TI - [The healing dynamics of periapical destructive lesions in x-ray imaging]. AB - Regeneration processes in periapical tissues were followed up after bone defect repair with hydroxyapatite-based osteoplastic preparations. The terms of regeneration depended on the size of bone defect. The data were evaluated by densitometry. PMID- 10812984 TI - [The use of nitatsid and gipozol'-N in the combined treatment of acute inflammatory diseases of the maxillofacial area]. AB - Combined drugs with a hydrophilic base, nitacide and hyposol-n, were used in combined therapy of acute purulent inflammations of the maxillofacial area. Time course of clinical parameters and changes in microflora were studied in patients administered different local treatments with different inflammatory reactions. The data indicate a high efficiency of the drugs, which optimized local therapy of suppurative wounds and are recommended for wide practical use. PMID- 10812985 TI - [The effect of ozone on the microbiological characteristics of the oral fluid in patients with mandibular fractures]. AB - Thirty-nine patients with mandibular fractures were treated with ozonized solutions. Qualitative and quantitative characteristics of oral fluid were studied and correlated to clinical data. PMID- 10812986 TI - [A method for treating mandibular defects with a complex autograft based on the trapezius muscle]. AB - Study of surgical anatomy of musculus trapezius with the adjacent skin and spine of the scapula helped determine the main neurovascular "port" of the muscle. The data were used for developing a method for treating mandibular defects with complex grafts on a mobile neurovascular pedicle. The site and port do not depend on sex but are related to age and constitution and determine the type of the transplant cut. PMID- 10812988 TI - [Combined nose deformities and the restoration of nasal breathing by a surgical method]. AB - Disorders in nasal respiration after posttraumatic combined deformations depend on deformation of the septum, pathological changes in the mucosa, and deformation of the external nose. Restoration of nasal breathing is determined by the type of surgical intervention. The nasal respiratory function recovered in 20 days to 5 months in the majority (47) of patients operated for traumatic combined deformations of the nose with impairment of nasal respiration. PMID- 10812987 TI - [The prevention of postoperative nose deformities following bone reconstructive operations on the maxillary complex]. AB - Prevention of postoperative deformation of the nose after bone repair surgery (transposition of the maxillary complex forward) is an important problem. The proposed method for osteotomy of the maxillary complex prevents or minimizes the postoperative deformation of the nasal cartilage after transposition of the maxillary complex. The method was used with good results in 35 patients with upper microretrognathia. PMID- 10812989 TI - [The surface structure of the mucosa of the normal denture bed and in patients with the sequelae of a cerebral stroke under the scanning electron microscope]. AB - Significant differences in the structure of the surface of prosthetic bed mucosa were revealed in healthy normal subjects and patients with after effects of cerebral stroke. The differences were observed both on the paralyzed and nonparalyzed side of prosthetic bed. PMID- 10812990 TI - [The composition and properties of the oral fluid in children taking sodium fluoride tablets who have different levels of oral hygiene]. AB - Oral hygiene of children aged 3-5 years and oral fluid composition and characteristics are directly related to each other. Treatment with sodium fluoride (orally) together with rational oral hygiene decreases salivary viscosity, values of enamel resistance test, modifies the rate of enamel remineralization, increases the Ca/P coefficient at the expense of decrease in the concentration of inorganic phosphorus, and increases fluorine concentration in the oral fluid. PMID- 10812991 TI - [A trial of performing dental caries prevention in preschoolers with fluoridated salt]. AB - Fluorinated salt was added to rations of preschool children for preventing dental caries. The program was implemented in 1411 children for 1 year, in 1438 for 2 years, in 1070 for 3 years, and in 1056 children for 4 years. The study was carried out in children attending kindergartens in Minsk, Mogilev, Mozyr, Nesvizh, and Baranovichi. Due to fluorinated salt, the incidence and intensity of caries decreased. In the group given fluorinated salt for 2 years, the incidence of caries decreased by 4.96% (p < 0.001), its intensity by the CDL (carious decayed-lost) index by 14.71%, and by the CPL (caries-prostheses-lost) index by 12.99%. Three-year consumption of fluorinated salt decreased the incidence of caries by 8.07% (p < 0.001), its intensity by the CDL index by 23.86%, and by the CPL index by 23.93%. In children using fluorinated salt for 4 years, these values decreased by 12.56 (p < 0.001), 32.80, and 29.64%, respectively. Hence, fluorinated salt can and should be used for preventing dental caries in collective bodies of preschool children. PMID- 10812993 TI - [The possible mechanisms and levels of quality control in dental care for the population]. AB - Mechanisms and levels of dental aid quality control are reviewed. The end point is the quality of treatment results because its most essential for the population. PMID- 10812992 TI - [The indices of caries and dental fluorosis prevalence in schoolchildren born and permanently living in regions of Azerbaijan differing by the level of the drinking water fluorine content]. AB - Epidemiological survey of schoolchildren born and permanently living in Baku (fluorine concentration in drinking water 0.3-0.5 mg/liter) and in the Azizbekov region at a distance of 30-40 km from Baku (fluorine concentration at least 1.5 mg/liter) was carried out. The incidence of dental diseases was estimated with consideration for environmental factors. High concentration of fluorine in drinking water in the Azizbekov region notably decreases the incidence of dental caries in the studied cohort of children. These data will"help improve organization and planning of dental service, specifically the treatment and prophylaxis measures for children. PMID- 10812994 TI - [The new-generation universal laser apparatus Optodan for the laser physio-, magneto- and reflexotherapy of stomatological diseases]. AB - Presents the design of a new-generation semiconductor laser with automated regulation (wavelength 0.85-0.95 micron, power up to 4 W, and frequency 0.1-3 kHz) and potentialities of its use for laser physio-, magneto-, and reflex therapy of dental and general somatic diseases in accordance with authors patented methods. PMID- 10812995 TI - [The diagnosis of syphilis]. PMID- 10812996 TI - [The risk factors and incidence of tooth loss in middle-aged and elderly persons]. PMID- 10812997 TI - [Academician of the Academy of Medical Sciences of the USSR Aleksei Alekssevich Minkh (1904-1984)]. PMID- 10812998 TI - [The history of the creation of drilling machines. 2]. PMID- 10813000 TI - Food microbiology. WHO-recognized course. PMID- 10812999 TI - WHO Eastern Mediterranean Region. Part II. Steps for effective forecasting. PMID- 10813001 TI - Immunology 2000. The American Association of Immunologists and Clinical Immunology Society Joint annual meeting. Seattle, Washington, USA. May 12-16, 2000. Abstracts. PMID- 10813002 TI - Digestive Disease Week and the 101st annual meeting of the American Gastroenterological Association. San Diego, California, USA. May 21-24, 2000. Abstracts. PMID- 10813003 TI - Digestive Disease Week and the 101st annual meeting of the American Gastroenterological Association. San Diego, California, USA. May 21-24, 2000. Abstracts. PMID- 10813004 TI - Tobacco in the courts. PMID- 10813005 TI - Treating stroke. PMID- 10813006 TI - Vitamin B12 injections for the elderly. PMID- 10813007 TI - Out of province, out of sight. PMID- 10813008 TI - Anaphylactoid reactions and angioedema during alteplase treatment of acute ischemic stroke. AB - Among 105 patients given recombinant tissue plasminogen activator (rt-PA, alteplase) intravenously for acute stroke, 2 (1.9%) had lingual angioedema, which progressed to a fatal anaphylactoid reaction in 1. The authors review the 2 cases and possible mechanisms responsible. They warn that patients who are taking an angiotensin-converting-enzyme inhibitor may be at increased risk for angioedema with concomitant alteplase therapy. PMID- 10813009 TI - Otolaryngologists' perceptions of the indications for tympanostomy tube insertion in children. AB - BACKGROUND: Bilateral myringotomy with insertion of tympanostomy tubes is the most common operation that children in Canada undergo. Area variations in surgical rates for this procedure have raised questions about indications used to decide about surgery. The objective of this study was to describe the factors that influence otolaryngologists to recommend tympanostomy tube insertion in children with otitis media and their level of agreement about indications for surgery. METHODS: A survey was sent to all 227 otolaryngologists in Ontario in the fall of 1996. The influence of 17 clinical and social factors on recommendations to insert tympanostomy tubes were assessed. Case vignettes were used to determine the effect of multiple factors in decisions about the need for surgical management. RESULTS: Surveys were returned by 138 (68.3%) of the 202 eligible otolaryngologists. There was agreement (more than 90% of respondents) about 6 indications for surgery: persistent effusion, a lack of improvement after 3 months of antibiotic therapy, a history of persistent effusion for 3 or more months per episode of otitis media, more than 7 episodes of otitis media in 6 months, a bilateral conductive hearing loss of 20 dB or more and a persistently abnormal tympanic membrane. Some respondents were more likely to recommend tube insertion if there were parental concerns about hearing problems or the frequency or severity of episodes of otitis media. Otolaryngologists agreed about the role of tympanostomy tubes in 1 of 4 case vignettes but disagreed about whether adenoidectomy should also be performed in that instance. Most viewed tympanostomy tube insertion as beneficial, with few adverse effects. INTERPRETATION: There is a lack of consensus among practising otolaryngologists in Ontario as to which children with recurrent otitis media or persistent effusion should undergo bilateral myringotomy with tympanostomy tube insertion. These findings suggest the need to revisit clinical guidelines for this procedure. PMID- 10813010 TI - Development and validation of the Osteoporosis Risk Assessment Instrument to facilitate selection of women for bone densitometry. AB - BACKGROUND: Although mass screening for osteoporosis is not recommended among postmenopausal women, there is no consensus on which women should undergo testing for low bone mineral density. The objective of this study was to develop and validate a clinical tool to help clinicians identify which women are at increased risk for osteoporosis and should therefore undergo further testing with bone densitometry. METHODS: Using Ontario baseline data from the Canadian Multicentre Osteoporosis Study, we identified all cognitively normal women aged 45 years or more who had undergone testing with dual-energy x-ray absorptiometry (DXA) at both the femoral neck and the lumbar spine (L1-L4). Participants who had a previous diagnosis of osteoporosis or were taking bone active medication other than ovarian hormones were excluded. The main outcome measure was low bone mineral density (T score of 2 or more standard deviations below the mean for young Canadian women) at either the femoral neck or the lumbar spine. Logistic regression analysis and receiver operating characteristic (ROC) analysis were used to identify the simplest algorithm that would identify women at increased risk for low bone mineral density. RESULTS: The study population comprised 1376 women, of whom 926 were allocated to the development of the tool and 450 to its validation. A simple algorithm based on age, weight and current estrogen use (yes or no) was developed. Validation of this 3-item Osteoporosis Risk Assessment Instrument (ORAI) showed that the tool had a sensitivity of 93.3% (95% confidence interval [CI] 86.3%-97.0%) and a specificity of 46.4% (95% CI 41.0%-51.8%) for selecting women with low bone mineral density. The sensitivity of the instrument for selecting women with osteoporosis was 94.4% (95% CI 83.7%-98.6%). Use of the ORAI represented a 38.7% reduction in DXA testing compared with screening all women in our study. INTERPRETATION: The ORAI accurately identifies the vast majority of women likely to have low bone mineral density and is effective in substantially decreasing the need for all women to undergo DXA testing. PMID- 10813011 TI - Ending waiting-list mismanagement: principles and practice. PMID- 10813012 TI - Why randomized controlled trials fail but needn't: a new series is launched. PMID- 10813014 TI - Why randomized controlled trials fail but needn't: 1. Failure to gain "coal-face" commitment and to use the uncertainty principle. PMID- 10813013 TI - Waiting for medical services in Canada: lots of heat, but little light. PMID- 10813015 TI - Defusing the intra-abdominal ticking bomb: intestinal malrotation in children. PMID- 10813017 TI - First Canadian live-donor lung transplants performed in Winnipeg. PMID- 10813018 TI - New and improved: CMA's guidelines infobase now at MDs' fingertips. PMID- 10813016 TI - Rheumatology: 3. Getting the most out of radiology. PMID- 10813019 TI - Female genital mutilation. PMID- 10813020 TI - Enter the hospitalist: new type of patient creating a new type of specialist. PMID- 10813021 TI - Canada's new doctors turning backs on family practice. PMID- 10813022 TI - Knowledge of cardiovascular disease risk factors among the Canadian population: relationships with indicators of socioeconomic status. AB - BACKGROUND: We examined the ability of adult Canadians to recall cardiovascular disease risk factors to determine the associations between their ability to recall risk factors for cardiovascular disease and their socioeconomic status. METHODS: This study used the database assembled by the Canadian Heart Health Surveys Research Group between 1986 and 1992--a stratified representative sample comprising 23,129 Canadian residents aged 18 to 74. Nurses administered a standard questionnaire asking respondents to list the major risk factors for cardiovascular disease: fat in food, smoking, lack of exercise, excess weight, elevated blood cholesterol and high blood pressure. Six logistic regressions examined the multivariate associations between ability to recall each risk factor with education, income adequacy, occupation, sex, age, marital status and province of residence. RESULTS: More people knew about the behaviour-related risk factors for cardiovascular disease than about the physiologic risk factors: 60% recalled fat in food, 52% smoking and 41% lack of exercise, but only 32% identified weight, 27% cholesterol and 22% high blood pressure. Education was the socioeconomic status indicator most strongly and consistently associated with the ability to recall risk factors for cardiovascular disease. The odds ratios of reporting an association of the risks between people with elementary education and those with university degrees varied between 0.16 (95% confidence interval 0.12 to 0.22) for lack of exercise to 0.55 (95% confidence interval 0.39 to 0.77) for smoking. INTERPRETATION: People in categories at greater risk of cardiovascular disease, such as those aged 65 or more or those with only elementary education, are less able to recall important cardiovascular disease risk factors. PMID- 10813024 TI - Protection of myocardial mitochondria against oxidative damage by selenium containing abzyme m4G3. AB - Selenium-containing abzyme (m4G3) was prepared and its protection of myocardial mitochondria against oxidative damage was studied using the swelling of mitochondria, quantity of lipid peroxidation products, and change in cytochrome-c oxidase activity as a measure of mitochondrial damage. The results showed that m4G3 could inhibit mitochondrial damage caused by the hypoxanthine-xanthine oxidase system in vitro. Electronic spin resonance (ESR) studies demonstrated that m4G3 could decrease the amount of free radicals generated in the damage system. PMID- 10813023 TI - Distribution of cardiovascular disease risk factors by socioeconomic status among Canadian adults. AB - BACKGROUND: This study was designed to describe the distribution of risk factors for cardiovascular disease by socioeconomic status in adult men and women across Canada using the Canadian Heart Health Surveys Database. METHODS: The data were derived from provincial cross-sectional surveys done between 1986 and 1992. Data were obtained through a home interview and a clinic visit using a probability sample of 29,855 men and women aged 18-74 years of whom 23,129 (77%) agreed to participate. The following risk factors for cardiovascular disease were considered: elevated total plasma cholesterol (greater than 5.2 mmol/L), regular current cigarette smoking (one or more daily), elevated diastolic or systolic blood pressure (140/90 mm Hg), overweight (body mass index and lack of leisure time physical activity [less than once a week in the last month]). Education and income adequacy were used as measures of socioeconomic status and mother tongue as a measure of cultural affiliation. RESULTS: For most of the risk factors examined, the prevalence of the risk factors was inversely related to socioeconomic status, but the relationship was stronger and more consistent for education than for income. The inverse relationship between socioeconomic status and the prevalence of the risk factors was particularly strong for smoking and overweight, where a gradient was observed: 46% (standard error [SE] 1.4) of men and 42% (SE 4.3) of women who had not completed secondary school were regular smokers, but only 12% (SE 1.0) of men and 13% (SE 0.9) of women with a university degree were regular smokers. Thirty-nine percent (SE 1.4) of men and 19% (SE 3.8) of women who had not completed secondary school were overweight, compared with 26% (SE 2.6) of male and 19% of female university graduates. The prevalence of leisure-time physical inactivity and elevated cholesterol was highest in both men and women in the lowest socioeconomic category, particularly by level of education. INTERPRETATION: The differences in the prevalence of risk factors for cardiovascular disease between socioeconomic groups are still important in Canada and should be considered in planning programs to reduce the morbidity and mortality from cardiovascular disease. PMID- 10813025 TI - Molecular analysis and heterologous expression of the gene encoding methylmalonyl coenzyme A mutase from rifamycin SV-producing strain Amycolatopsis mediterranei U32. AB - The conversion of succinyl-coenzyme A (CoA) into methylmalonyl-CoA, catalyzed by adenosylcobalamin-dependent methylmalonyl-CoA mutase (MCM), represents an important source of building blocks for rifamycin SV biosynthesis. The structural gene for MCM from rifamycin SV-producing strain Amycolatopsis mediterranei U32 was isolated by using a heterologous gene probe encoding the MCM of Streptomyces cinnamonesis. A 7.8-kbp fragment was sequenced and four complete open reading frames (ORFs) and two incomplete ORFs were found. Two central ORFs, ORF3 and ORF4, overlap by four nucleotides and were found to encode MCM small (602 residues) and large (721 residues) subunits, respectively. Comparison showed that the MCM gene of A. mediterranei U32 was quite similar to those from other sources. The functionally unknown ORF5, immediately downstream of the mutAB gene, was quite similar to the ORFs downstream of mutAB from S. cinnamonensis and Mycobacterium tuberculosis. Such a striking cross-species conservation of gene order suggested that ORF5 could also be involved in the metabolism of methylmalonyl-CoA. MCM gene was overexpressed in Escherichia coli under T7 promoter, and MCM activity could be detected in the recombinant E. coli clone harboring MCM gene after the addition of coenzyme B12. A purification procedure based on the B12 affinity column was established to purify the MCM from E. coli. The molecular weight of purified MCM from E. coli was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, which corresponds to that calculated from the MCM protein sequence and is also the same size as that of the enzyme purified directly from A. mediterranei U32. MCM gene was overexpressed in polyketide monensin producing S. cinnamonensis, and the total monensin production was increased by 32%. PMID- 10813026 TI - Catalytic and spectroscopic properties of cytochrome-c, horseradish peroxidase, and ascorbate oxidase embedded in a sol-gel silica matrix as a function of gelation time. AB - In this study, we investigated the optical features of the redox metal-dependent proteins cytochrome-c, horseradish peroxidase (HRP), and ascorbate oxidase embedded in a sol-gel-processed silica matrix as a function of gelation time. Circular dichroism, absorbance, and fluorescence spectroscopies revealed that the sol-gel process affects the complex structure of the dimeric ascorbate oxidase (although the prosthetic coppers still remain bound to the enzyme) but not that of monomeric cytochrome-c and HRP. Any modifications in ascorbate oxidase occurred in the initial gelation phase; the drying process induced no further alterations and the enzyme remained stable for months. Unfolding-refolding experiments on cytochrome-c revealed severely restricted motility in the protein moiety in the xerogel, the concentrated matrix that forms after drying. The diffusion time of the solvent within the matrix, which regulated the enzyme substrate reaction rate, depended on the thickness of the monolith, not on the dryness of the specimen. PMID- 10813027 TI - Stable isotopes in routine clinical use. PMID- 10813028 TI - Comparison between the oral and intravenous L-[1-13C]phenylalanine breath test for the assessment of liver function. AB - To simplify the L-[1-13C]phenylalanine breath test which is used to assess liver function the tracer is usually given orally, and CO2 production rate is estimated. In 12 healthy volunteers and 10 liver cirrhotics we compared the oral approach with i.v. tracer administration combined with measurement of individual CO2 production rate. The 13CO2/12CO2 enrichment was assessed by isotope-ratio mass spectrometry. After i.v. [1-13C]phenylalanine application exhaled 13C recovery per minute peaked within 10 minutes (controls: 0.17 +/- 0.06%; cirrhotics: 0.05 +/- 0.02%, p < 0.01). The oral approach yielded comparable separation between 30-60 minutes, with average peak values being 0.18 +/- 0.03% and 0.06 +/- 0.03% (p < 0.01), respectively. Variable gastrointestinal resorption kinetics after oral application probably causes this difference. PMID- 10813029 TI - Transfusing blood: supply and demand. PMID- 10813030 TI - Viral folliculitis. AB - Viral folliculitis is an infrequently reported entity. We describe two patients with viral folliculitides, including a case of herpetic sycosis caused by herpes simplex (HSV) and a case of herpes zoster (HZ) without blisters. Clinicians should consider viral etiologies in the differential diagnosis of superficial infectious folliculitis, especially those cases refractory to antibacterial or antifungal therapy. PMID- 10813031 TI - The use of antiretroviral agents in patients with renal insufficiency. AB - Current guidelines for the dosing of antiretroviral agents in HIV-positive patients with renal insufficiency including those requiring dialysis are limited, as revealed by a literature search performed using Health Knowledge Network MEDLINE and HealthSTAR. Pharmacokinetic and chemical aspects were taken into consideration to develop further recommendations. Results showed reverse transcriptase inhibitor dosage adjustment guidelines are available, but for patients receiving dialysis, guidelines are limited to zidovudine and didanosine. There are limited clinical data on dosage adjustments required for the use of protease inhibitors or non-nucleoside reverse transcriptase inhibitors (NNRTIs). Indinavir appears to require no dose modification. For nelfinavir, ritonavir and saquinavir, the chemical and pharmacokinetic characteristics along with the pathophysiology of renal dysfunction and dialysis suggest that these agents can likely be given at usual doses with close monitoring for toxicities. More data is required to make recommendations for use of NNRTIs in renal insufficiency. Combining the information from previous dosing guidelines and the pharmacokinetic data of antiretroviral agents allows for preliminary recommendations to be made until confirmed by definitive clinical data. PMID- 10813032 TI - HIV-associated malignancies in children. AB - The exact incidence of malignancies in HIV-infected children is not clear; however, an excess of non-Hodgkin's lymphomas (NHLs) and soft tissue tumors (leiomyosarcomas) is evident. The spectrum of diseases is slightly different in children compared to adults. For example, Kaposi's sarcoma (KS), although common in HIV-infected adults, is rare in children in industrialized countries. Preliminary results with brief, dose-intensive chemotherapeutic regimens have been encouraging. Such regimens may be complicated, however, by multiple HIV associated organ dysfunctions, drug interactions, and infectious complications secondary to severe immunosuppression. PMID- 10813033 TI - Barriers to adherence to highly active antiretroviral therapy as expressed by people living with HIV/AIDS. AB - The primary objective of this study was to gain a clearer understanding of the barriers to adherence to highly active antiretroviral therapy (HAART) faced by people living with HIV/AIDS (PLWHIV/AIDS) on Long Island, New York. Focus group, a qualitative research method, was used to study these barriers. The study was conducted in 1998 on Long Island, NY, at five institutions that provide services to 1700 PLWHIV/AIDS. Five focus groups were conducted with 6 to 13 PLWHIV/AIDS in each group, a total of 39 subjects. PLWHIV/AIDS identified eight common barriers to adherence to HAART. In descending order, the barriers include: (1) frequency and severity of side effects, (2) conflicts with daily routines, (3) dietary requirements, (4) frequency of taking medications, (5) number and dosage of medications, (6) psychosocial factors (i.e., stress, feeling good, and bad news), (7) pharmacy refills, and (8) physiological needs (i.e., sleep, hunger, or thirst). Many factors play a role in the success or failure of HAART, including preexisting drug resistance, drug-drug interactions, and the ability of PLWHIV/AIDS to adhere to a rigid and frequently changing medication regimen. The information gleaned from focus groups is limited in that it may not be generalized to a larger population with any known reliability. However, clinicians sensitive to barriers to adherence to HAART, including those identified by PLWHIV/AIDS in this study, may play a more proactive role in supporting adherence to the medication regimen, increasing the durability of effective viral suppression, decreasing morbidity and mortality, and decreasing the selection and transmission of resistant strains of HIV. PMID- 10813034 TI - Talking about AIDS. AB - Despite increased public and professional awareness, patients and physicians tend to avoid discussions about HIV. Empiric studies of patient-physician communication point to specific common communication breakdowns. These include lack of a good opening line, inappropriate context, awkward moments, vague language, and a physician-centered rather than a patient-centered interview style. In effective HIV-related discussions, clinicians elicit the patient's beliefs and concerns first, are organized, use empathy, provide a rationale for the discussion, persist through awkward moments, and clarify vague language. In addition to information about sexual behaviors and the number, gender, and HIV status of partners, clinicians should ask about the context and antecedents to risk behaviors, praise prior attempts to reduce risk, and assess the patient's motivation to change. Although studies indicate that experienced practitioners often do not have these skills, they can be learned. PMID- 10813035 TI - Psychological discomfort and mental illness in patients with AIDS: implications for home care. AB - A chart review study of 100 consecutive AIDS patients cared for by our Home Care (HC) unit was performed. The main characteristics of the sample were: M/F ratio = 3/1; mean age = 39.7 +/- 6.6; mode of transmission = 56% intravenous drug users (IVDU); Centers For Disease Control (CDC) classification = 98% C3; mean length of HC = 5.2 months. Psychopathological evaluation led to an overall rate of morbidity during HC of 73%. The most common psychiatric diagnoses were dementia and adjustment disorders. Nearly two thirds of the patients received psychopharmacological treatment; one fifth were still on methadone. Psychological discomfort was perceived by more than half of patients. Interpersonal problems with cohabitants were noticed by caregivers in 41%. Female patients showed an increased prevalence of anxiety and depressive disorders. IVDUs had more prominent psychosocial problems. Interpersonal problems with cohabitants were correlated to substance use or other mental disorders in the family and to the patient's psychological discomfort. We conclude that mental health professional intervention for AIDS patients is strongly needed in a home-care setting. PMID- 10813036 TI - HIV/AIDS case histories: diagnostic problems. Cryptococcoma of the lung. PMID- 10813037 TI - NDA submitted for adefovir. PMID- 10813039 TI - Ziagen cleared for Europe. PMID- 10813038 TI - Norvir capsule approved. PMID- 10813040 TI - Phase II begins for T-20. PMID- 10813041 TI - New PI active against many strains. PMID- 10813043 TI - Metabolic profile of protease inhibitors studied. PMID- 10813042 TI - LEUKINE maintains viral suppression and extends duration of antiretroviral therapy. PMID- 10813044 TI - PI trial seeks participants. PMID- 10813045 TI - HCV rate high in Baltimore. PMID- 10813046 TI - Syphilis rates drop among teens. PMID- 10813047 TI - Lack of volunteers for vaccine. PMID- 10813048 TI - Cervical vaccine promising. PMID- 10813049 TI - Coconut oil compound may treat STDs. PMID- 10813050 TI - Prevention programs producing positive results. PMID- 10813051 TI - Sexual risk behaviors decline among high schoolers. PMID- 10813052 TI - Community and school HIV prevention programs help. PMID- 10813053 TI - Intensive control programs lead to TB decline. PMID- 10813054 TI - [II Brazilian Meeting of Researchers on Alzheimer's Disease and Related Disorders. Abstracts]. PMID- 10813055 TI - The 13th International Conference on Antiviral Research. Baltimore, Maryland, USA. April 16-21, 2000. Abstracts. PMID- 10813056 TI - Society of Biological Psychiatry 55th annual meeting. Chicago, Illinois, USA. May 11-13, 2000. Abstracts. PMID- 10813057 TI - British Society of Audiology Short Papers Meeting on Experimental Studies of Hearing and Deafness. University of Essex, 21-22 September 1999. Abstracts. PMID- 10813058 TI - XXVII European Symposium on Calcified Tissues. Tampere, Finland, 6-10 May 2000. Abstracts. PMID- 10813059 TI - European Congress of the International Hepato-Pancreato-Biliary Association (IHPBA). Budapest, Hungary, May 24-28, 1999. Abstracts. PMID- 10813060 TI - 1st European College of Neuropsychopharmacology (ECNP) Workshop. Nice, France, March 16-19, 2000. Abstracts. PMID- 10813061 TI - 5th European College of Neuropsychopharmacology (ECNP) regional meeting. St. Petersburg, Russia. April 15-17, 2000. Abstracts. PMID- 10813063 TI - [Neurology Meeting of the French Language. Montpellier, France, 12-15 April 2000. Abstracts]. PMID- 10813062 TI - 2000 Annual Meeting and 40th Symposia of the Japanese Society of Plant Physiologists. March 27-29, 2000. Abstracts. PMID- 10813064 TI - [Satellite Symposia of the Neurology Meeting of the French Language. Montpellier, France, 12-15 April 2000]. PMID- 10813065 TI - [66th Annual meeting of the German Society for Cardiology, Heart, and Circulatory Research. Mannheim, 27-29 April 2000. Abstracts]. PMID- 10813066 TI - Time for evaluation of early intervention and prevention in major psychiatric disorders. PMID- 10813067 TI - Toward effective early intervention and prevention strategies for major affective disorders: a review of antecedents and risk factors. AB - OBJECTIVE: To review critically the literature pertaining to risk factors and antecedent symptoms and syndromes in order to determine an empirically based strategy for early treatment and prevention of major mood episodes. METHOD: The relevant literature is summarized, with particular emphasis on early-onset (child and adolescent) mood disorders. RESULTS: A complex interaction between biological, psychological, and sociological factors contributes to the development of a major mood disorder. Having a positive family history of mood disorder (bipolar and unipolar) and being female (unipolar) are the strongest, most reliable risk factors. There is continuity between adolescent and adult mood disorders, and subsyndromal mood disturbance in adolescents has clinical and public health significance. However, more longitudinal study is required to reliably map the course and predictive importance of mood disorders in very young children. CONCLUSIONS: Substantial evidence supports the effectiveness of early intervention and prevention efforts in children at risk for mood disorders (identified as having affected family members) and in adolescents manifesting significant mood symptoms and syndromes (especially if associated with a positive family history). However, the current level of understanding regarding the etiological significance and mechanism of risk factors associated with mood disorders does not support broad community-based primary prevention strategies in unselected populations. PMID- 10813068 TI - Staff feelings and patient diagnosis. AB - OBJECTIVE: To assess the associations between staff feelings toward patients and the patients' diagnoses, in view of the fact that clinical reports of such associations have not been corroborated by systematic research. METHOD: At 24 psychiatric units, 143 patients were assessed according to their personality organization, and staff feelings toward these patients were followed for 5 years. The feelings were reported on a feeling checklist twice yearly, and outcome was assessed as the effect size at year 5, using ratings on Kernberg's structural model complemented with ratings on Strauss-Carpenter's function scale. RESULTS: The study showed that it was possible, using discriminant analyses, to separate diagnostic groups by the different feelings that they evoked in the staff. Patients with borderline personality organization (BPO) evoked fewer relaxed and more aggressive feelings, in contrast to patients with psychotic personality organization (PPO). In contrast to patients with neurotic personality organization (NPO), who evoked feelings of sympathy and helpfulness, PPO patients evoked more feelings of insufficiency and disappointment. A stepwise discriminant analysis of reactions to patients with positive treatment outcome separated the 3 personality organizations with 2 functions using only 2 feelings, "relaxed" and "objective." The feeling relaxed separated the NPO patients from the BPO patients, and the feeling objective separated the PPO patients from the other groups. The patients' diagnoses accounted for larger proportions of variance in feelings for the patients with positive outcome. CONCLUSION: The results implied that the patients' different personality organizations evoked different staff feelings in this treatment context and that positive treatment outcome was associated with more pronounced and clear-cut staff reactions. PMID- 10813069 TI - Psychiatric disorders in an Arctic community. AB - OBJECTIVE: To determine the rates of depression, anxiety, and alcohol abuse, using modern nosology, in a random sample of residents aged 14 to 85 years living in an Arctic community. METHOD: A cross-sectional 2-step survey of randomly selected households was undertaken, using a self-report questionnaire to screen for anxiety, depression, and alcohol abuse. The survey included the Hospital Anxiety and Depression Scale (HADS) and Ewing and Roose's 4-question alcohol screening instrument (the CAGE questionnaire). Cut-off scores for the HADS and CAGE were found by comparing HADS and CAGE scores with scores on the Structured Clinical Interview for the DSM-III-R (SCID) in a stratified subsample. RESULTS: Estimated rates of depression and anxiety were 26.5% and 19.0% respectively within the past week, and estimated rates of lifetime alcohol abuse were 30.5%. CONCLUSIONS: The estimated prevalence of psychiatric disorders in this Arctic community is higher than that indicated in previous findings on Native mental health. PMID- 10813071 TI - A longitudinal study of siblings of children with chronic disabilities. AB - OBJECTIVE: To examine the unaffected siblings of 2 different groups with chronic disabilities, pervasive developmental disorder (PDD) and Down syndrome (DS), over 3 years, comparing their adjustment with each other and with the siblings of a nondisabled group. METHOD: This study examines 137 siblings of children with PDD, children with DS, and developmentally normal children (control group) initially and 127 siblings at follow-up 3 years later. Their adjustment is measured by the Survey Diagnostic Instrument (SDI), completed by caregivers and teachers. Predictor variables include sibling self-perception, social support, and relationship with sibling, as indicated by siblings; caregiver psychosocial factors such as parental stress, caregiver depression, and marital relationship; family systems characteristics as viewed by both caregiver and sibling; and difficulty that disabled child causes as perceived by the primary caregiver. RESULTS: Significantly more adjustment problems are found in the siblings of PDD children at both times when compared with siblings of DS and control children. Caregivers of PDD children report the highest levels of distress and depression, and this persists over time. Parent distress was found, at both times, to be related to sibling adjustment problems, regardless of study group. CONCLUSION: These results have implications for preventive intervention for the unaffected siblings of PDD children. PMID- 10813070 TI - [Deficit of the theory-of-mind in schizophrenia: clinical concept and review of experimental arguments]. AB - OBJECTIVE: This article synthesizes and discusses the clinical and experimental arguments for applying the concept of a theory-of-mind deficit to schizophrenia. METHOD: The term "theory of mind" is defined. A review of literature on the hypothesis of a theory-of-mind deficit in schizophrenia is then presented. Clinical and experimental arguments are discussed. The article concludes with a summary of the issue. RESULTS: The hypothesis of a theory-of-mind deficit in schizophrenia is convincing, particularly considering the most recent experimental evidence. This cognitive deficit explains the presence of schizophrenia symptoms such as delusions of influence and reference, problems in social adaptation, and signs of disorganized communication. Consideration is given to the roles played by situational ambiguity, the individual's intellectual level, and type of treatment. CONCLUSION: The hypothesis of a theory-of-mind deficit has only recently been applied in schizophrenia research; it may have a strong impact on clinical practice. If its role in the onset of symptoms is to be better understood, explanatory models and experimental paradigms must be further refined. PMID- 10813073 TI - Natural disasters and service delivery to individuals with severe mental illness- ice storm 1998. AB - OBJECTIVE: To review the literature on the responses of individuals with severe mental illness (SMI) to natural disasters, to describe the impact of the 1998 Ice Storm on a group of SMI patients, and to describe the steps taken at a Canadian university teaching hospital to ensure the ongoing provision of mental health services throughout the crisis. METHOD: Published articles describing the impact of natural disasters on SMI populations, as well as service provision to these patients, are reviewed. Service use at the Montreal General Hospital (MGH) Department of Psychiatry is described. A questionnaire about the impact of the ice storm was administered to a group of patients in an assertive community treatment program. RESULTS: Service use during this natural disaster was consistent with that described in the literature, in that these patients were no more likely to be admitted or to visit the emergency room during the crisis. Continuous mental health service delivery may have contributed to this positive outcome. This service delivery was provided by ensuring staff access to information, by securing the physical safety of both staff and patients, and by taking a flexible, outreach-oriented approach to service delivery. CONCLUSIONS: SMI patients who have ongoing access to psychiatric services in disaster situations tend to cope well. A flexible, proactive, assertive approach to service delivery during the crisis situation will help to ensure that needs for care will be met. PMID- 10813072 TI - Inpatient treatment of severe disruptive behaviour disorders with risperidone and milieu therapy. AB - OBJECTIVES: To evaluate the therapeutic impact of adding risperidone to milieu therapy of latency-aged inpatients with severe disruptive disorders. METHODS: The charts of 90 latency-aged patients consecutively admitted to a psychiatry ward were reviewed retrospectively. Fifteen of these patients received risperidone treatment, were nonpsychotic, and did not suffer from pervasive developmental disorder (12 male, 3 female; mean age 9.99 years, SD 1.76). Their scores on the Children's Global Assessment Scale (CGAS) were compared at admission, before risperidone treatment, and at discharge. RESULTS: All subjects were diagnosed with a disruptive behavioural disorder. Ten (66.67%) had additional learning difficulties, and 13 (86.7%) had pathological personality traits. The characteristics of the sample suggested borderline pathology or multiple complex developmental disorder. Following a mean of 38 days after admission (SD 22.3), the patients received risperidone for a mean of 46 days (SD 28.2) before being discharged. The mean maintenance dose of risperidone was 1.27 mg daily (SD 0.36). Mean CGAS scores increased from admission (21.9, SD 7.0) to before risperidone treatment (26.8, SD 7.6, P < 0.0001) and to discharge (50.3, SD 5.3, P < 0.0001). Only 2 patients had documented side effects. CONCLUSIONS: Low-dose risperidone used adjunctively to milieu therapy led to statistically and clinically significant additional improvement in the functioning of hospitalized latency aged children with severe behavioural disorders. Low-dose risperidone is a safe and effective adjunct to milieu therapy for treating this population in inpatient settings. Prospective randomized controlled trials are needed to confirm these findings. PMID- 10813074 TI - Criteria for admission to psychiatric units: have we raised the bar too high? PMID- 10813075 TI - Olanzapine treatment of obsessive-compulsive disorder. PMID- 10813076 TI - Clozapine--an atypical reaction. PMID- 10813077 TI - Clozapine treatment and neuroleptic malignant syndrome. PMID- 10813078 TI - Clozapine therapy and pulmonary embolism. PMID- 10813079 TI - Another idea for the treatment of patients with borderline personality disorder. AB - We are not suggesting that casting is appropriate for all patients with BPD, nor are we suggesting that patients need be casted for prolonged periods of time. In the 3 stated cases, the patients were desperate to have some behaviour modification introduced into their lives and welcomed the procedure. In all these cases, comorbidity existed. Because marked skin damage and open wounds often occur, other specialists with expertise in wound healing and joint mobility maintenance need to be involved. We were fortunate to have the services of a plastic surgeon with a special interest in psychiatric disorders. We believe that psychiatrists should be aware of this technique as a possible strategy to be used in very specific circumstances. It appears, in our short series of 3 cases, to have benefits in very specific cases. If it appears that casting may be helpful, our recommendations are as follows: 1. The behaviours should be thoroughly evaluated, and all alternative managements should be reviewed. 2. Reasonable rapport needs to be established between the patient and the major therapists so that compliance can be assured. 3. Other specialists should be consulted, such as plastic surgeons, who have the expertise to manage prolonged casting. 4. Along with casting, appropriate medications and psychotherapy must be in place for a balanced approach to management. PMID- 10813080 TI - Re: Is hospitalization bad? PMID- 10813081 TI - Risperidone-induced obsessive-compulsive symptoms: a reappraisal. PMID- 10813082 TI - Paroxetine and tardive akathisia. PMID- 10813083 TI - A preventive services program model for preserving and supporting families over time. AB - Selected findings from a three-year study are reported regarding a prospective sample of 189 families served by the Center for Family Life's preventive services program and the nature and results of the services the families received. The program combines elements of both family preservation and family support services to provide a comprehensive, individualized response to families in need and prevent the unnecessary placement of children in care. Four program elements correspond with those typically identified as characterizing family preservation programs, and three key program elements differentiate the Center's approach from other family preservation programs. These latter characteristics are more typically found in family support programs and address limitations of current family preservation programs as identified in the literature. PMID- 10813084 TI - Power through choices: the development of a sexuality education curriculum for youths in out-of-home care. AB - Youths in out-of-home care demonstrate high rates of sexual risk-taking behavior and elevated rates of unintended pregnancy and sexually transmitted infections (STIs). This article profiles the development and characteristics of an innovative pregnancy/HIV/STI prevention curriculum tailored to the needs of youths in out-of-home care. Promising results from an implementation study suggest the need for further dissemination and rigorous testing. PMID- 10813085 TI - Attitudes of low-income parents toward seeking help with parenting: implications for practice. AB - Fifty-two low-income parents were surveyed to determine attitudes toward parenting and help seeking. Although a majority agreed that most parents, even "good" parents, need help or advice about parenting and thought they would seek help with parenting, low-income parents were less likely to believe in or seek out help than those with higher incomes. The most frequently selected sources of help were family, books and videos, telephone help-lines, and friends. The least likely sources of help were child protective services, school personnel, clergy, and social service/counseling agencies. Parent support and education groups were likely sources of support for only one in four low-income parents. PMID- 10813086 TI - The heterogeneity of children and their experiences in kinship care. AB - Increasingly, children in need of out-of-home care are being placed in kinship care, yet few studies have followed their placement histories longitudinally to determine if these children constitute a homogeneous group or heterogeneous subgroups. This study of 484 children in kinship care in San Diego County, California, indicates that children in kinship care have markedly different sociodemographic and maltreatment histories, as well as heterogeneous placement experiences. PMID- 10813087 TI - Presence and quantification of neuropeptide Y in pulmonary edema fluids in rats. AB - The present study was undertaken to determine the contribution of neuropeptide Y to edema occurrence in neurogenic and hydrostatic pulmonary edema. To induce neurogenic pulmonary edema, fibrinogen and thrombin were injected into the cisterna magna; and to evoke hydrostatic pulmonary edema, saline was infused intravenously. Concentrations of neuropeptide Y in serum and edema fluid were measured using enzyme-linked immunosorbent assay (ELISA), which showed a mean value of 158 nM (95% confidence limit 124-202 nM) in the neurogenic edema fluid, significantly higher than that in the hydrostatic one. Using immunohistochemistry, fluorescent signals reactive to neuropeptide Y were found in the alveolar macrophages and edema fluid in case of fibrin-induced pulmonary edema, but were almost absent in hydrostatic edema and absent in normal lungs. Mean ratio of protein concentrations in edema fluid to that in serum was 74.9 +/- 0.9% in fibrin-induced pulmonary edema, being higher than that in hydrostatic one. From these results, we conclude that neuropeptide Y has a relationship to the high protein concentration ratio, i.e., to increased pulmonary vascular permeability, and consequently may contribute to the development of neurogenic pulmonary edema in rats. PMID- 10813088 TI - Prolonged airway retention of insoluble particles in cystic fibrosis versus primary ciliary dyskinesia. AB - Patients with cystic fibrosis (CF) and primary ciliary dyskinesia (PCD) have been shown to have impaired large airway clearance of radiolabelled particles as measured by external gamma camera scanning up to 6 hours post deposition. Recent studies suggest that 24-hour retention of particles may reflect some airway retention in addition to alveolar retention. In a retrospective study, we analyzed the relationship between the deposition pattern and 24-hour retention (Ret24 hr) of technetium 99-radiolabelled iron oxide (99Tc-Fe2O3) particles in 20 patients with CF, 12 patients with PCD, and 17 normal subjects. By gamma camera analysis, initial aerosol deposition was analyzed in terms of central-peripheral (C/P) activity within the lungs. Gamma camera scanning was performed immediately following deposition and again at 24 hours to assess residual retention (Ret24 hr) as a percent of initial deposition. C/P analysis was also performed on the 24 hour scan (C/P24). For all subjects, initial deposition pattern (C/P) was inversely related to lung function (forced expiratory volume in 1 second [FEV1]%pred vs. C/P, r = -.54). Ret24 hr was also inversely related to initial deposition pattern for all subjects (Ret24 hr vs. C/P ratio, r = -.42). Analysis of covariance showed that for a given C/P ratio, CF patients had significantly greater Ret24 hr compared to normal subjects (9.8 +/- 2.8 [SE]%). In addition, the CF patients had similar C/P24 as the normal subjects (1.35 +/- 0.40 [SD] vs. 1.10 +/- 0.39, respectively). These results suggest that small airway clearance is compromised in CF patients compared to normal subjects. On the other hand, PCD patients had C/P24 similar to their initial deposition C/P ratios (2.78 +/- 1.72 vs. 2.45 +/- 0.87, respectively), significantly greater than 1.0, and significantly greater than CF or normal subjects, suggesting that PCD patients have prolonged particle retention associated with their large bronchial airways. PMID- 10813089 TI - The effect of alginate on the invasion of cystic fibrosis respiratory epithelial cells by clinical isolates of Pseudomonas aeruginosa. AB - Chronic infection in the cystic fibrosis (CF) lung is characterized by Pseudomonas aeruginosa strains that overproduce the mucoid exopolysaccharide, alginate. Previous experiments have shown that long-term survival of P. aeruginosa in the CF lung may be facilitated by increased adherence and decreased invasion of respiratory epithelial cells. Therefore, mucoid and nonmucoid clinical isolates of P. aeruginosa were assayed for their ability to associate with and invade the CF respiratory epithelial cell line, CF/T43. Association assays and gentamicin exclusion assays demonstrated that mucoid P. aeruginosa associates with and invades CF/T43 cell monolayers significantly less than nonmucoid P. aeruginosa strains (P = .004, .02). Fluorescence microscopy invasion assays confirmed these results. The differences in association and invasion by the P. aeruginosa strains were not due to differences in lipopolysaccharide phenotype or cytotoxicity for CF/T43 respiratory epithelial cells. Exogenous bacterial alginate had no effect on the invasion of CF respiratory epithelia by a nonmucoid strain. Invasion assays with the wild-type P. aeruginosa strain PAO1 and isogenic algU and mucA mutant strains failed to show differences in invasion (P = .25). We conclude that (i) mucoid P. aeruginosa isolates associate with and invade CF/T43 respiratory epithelial cells with less efficiency than nonmucoid P. aeruginosa, (ii) these differences are not due to variations in lipopolysaccharide phenotype between strains, (iii) neither exogenous nor endogenous alginate affects the ability of P. aeruginosa to invade CF/T43 respiratory epithelial cells, and (iv) invasion of CF/T43 respiratory epithelial cells by a laboratory reference strain of P. aeruginosa does not appear to be regulated by AlgU. PMID- 10813090 TI - Enhancement of liposome-mediated gene transfer to human airway epithelial cells by replication-deficient adenovirus. AB - We hypothesized that replication-deficient adenovirus (Ad), when complexed with plasmid DNA (pl) and cationic liposomes (L), would enhance liposome-mediated gene transfer in cultured human airway epithelial cells. Pl/L/Ad complexes were formed using charge-charge interactions. A gel electrophoresis retardation assay showed plasmid DNA to be associated with the virus in a high-molecular-weight, low mobility complex, the diameter of which was 300 to 350 nm. Compared to pl/L alone, pl/L/Ad enhanced luciferase expression on average by 1 log-fold in human airway epithelial cells which express either mutant or wild-type cystic fibrosis transmembrane conductance regulator (CFTR). Transgene expression was sustained at high levels for up to 7 days following transfection with pl/L/Ad. Using a heat stable alkaline phosphatase reporter gene, we showed that a larger fraction of cells was transfected by pl/L/Ad compared to pl/L. Finally, cells were exposed to Ad for 0 to 24 hours prior to pl/L or exposed to pl/L prior to Ad. We found that enhancement was significantly greater using pl/L/Ad compared to the simultaneous addition of Ad with the pl/L complexes. In addition, when pl/L was added 4 to 24 hours prior to Ad, some enhancement was found, suggesting that plasmid DNA remained in a compartment in the cell for several hours and became available for transcription with the addition of Ad. When Ad was added prior to pl/L, enhancement was found suggesting that the effect of the virus on cell membranes may persist for up to 24 hours. We conclude that the tripartite pl/L/Ad complex significantly enhances liposome-mediated transgene expression for a prolonged period of time in human bronchial epithelial cells. PMID- 10813091 TI - Hemoglobin potentiates the production of reactive oxygen species by alveolar macrophages. AB - The objectives of this investigation were (1) to determine the effects of hemoglobin on the production of reactive oxygen species by activated rat alveolar macrophages, (2) to determine a possible mechanism for these effects, and (3) to determine which part of the hemoglobin molecule is responsible for these effects. Production of reactive oxygen species by phorbol myristate acetate (PMA) stimulated cells was assessed by measuring luminol-enhanced chemiluminescence (CL). Hemoglobin enhances PMA-stimulated CL in a dose-dependent manner. The effect is maximal at 0.5-1.0 microM hemoglobin where PMA-induced CL is increased by approximately 20-fold. Superoxide anion release from PMA-stimulated cells is not affected by hemoglobin. However, the hemoglobin-induced enhancement of PMA stimulated CL is inhibited by superoxide dismutase, catalase, dimethylthiourea, or deferoxamine. These results suggest that hydroxyl radical may be formed from hydrogen peroxide which is derived from superoxide anion. Measurements of electron spin resonance spectra following spin trapping of radicals verify that hydroxyl radicals are produced by the cells in the presence of PMA and hemoglobin. The hemoglobin effects appear to require iron in a protoporphyrin complex, because hemin stimulates PMA-induced CL, whereas neither ferrous nor ferric iron has any effect. These findings taken together suggest that hemoglobin can act as a biological Fenton reagent to enhance the production of reactive oxygen species from alveolar macrophages and potentially contribute to lung damage during leakage of blood into the alveolar spaces. PMID- 10813092 TI - Immunolocalization of the boar seminal immunosuppressive fraction infused via uterus on the lymphocytes populating mouse genital tract tissues. AB - Intrauterine deposition of the immunosuppressive fraction from boar seminal vesicle fluid (ISF) led to a suppression of antibody response to soluble and corpuscular antigens in mice. By means of an immunofluorescent method using specific monoclonal antibody, ISF was detected on the membranes of white blood cells and splenocytes of mice subjected to intrauterine treatment from the third day to the thirteenth day after its deposition. ISF was also detected on the lymphocytes populating the mucosal tissues of vagina, cervix, oviduct and uterus from day 1 to 13 after its intrauterine administration. The antibody to soluble and corpuscular antigens was inhibited in the mice treated with ISF, but after the cessation of the ISF application, a normal immune response was restored within 40 days. Sandwich immunosorbent assay revealed that intrauterine infusion of ISF decreased significantly the concentration of IgG and IgM in the sera of immunized mice both after the primary and the secondary immunizations. These findings indicate that the intrauterine infusion of semen may influence the immune defense reactions and may be an important factor in the development of viral and bacterial infections of the female reproductive tract. PMID- 10813093 TI - Melatonin and retinyl acetate as chemopreventives in DMBA-induced mammary carcinogenesis in female Sprague-Dawley rats. AB - One goal of experimental oncology is to find and test effective chemopreventive substances which can suppress malignant transformation of the cells, their accumulation and invasion. Mammary gland tumours were induced by DMBA applied intragastrically (10 mg/rat, three times) every three days between postnatal days 50 and 60 in female Sprague-Dawley rats. One day after the last dose we started chemoprevention with Mel, RA and combination of both drugs, which lasted 25 weeks. Mel was drunk continuously as a solution in tap water (100 micrograms/ml). RA was applied daily in the dose of 8.2 mg/rat at the base of the tongue. There were four experimental groups: 1. control--no chemoprevention, 2. Mel treatment, 3. RA treatment, 4. application RA + Mel. At the end of the experiment the incidence, frequency, latency and average volume of tumours were evaluated. In the group treated with Mel tumour incidence, latency and volume did not differ from controls; the frequency of tumours was decreased. Treatment with RA and with combination RA + Mel decreased mammary tumour incidence to 38% (RA) and 48% (RA + Mel); it also decreased frequency and prolonged latency. Thus chemoprotective effects of RA and combination of RA with Mel were proved in mammary carcinogenesis induced by DMBA. The oncostatic effect of Mel alone was not confirmed. In our recent paper (Bojkova et al., 2000) drinking of lower doses of Mel during the late afternoon and night prolonged the latency period and in combination with RA showed an oncostatic effect on mammary carcinogenesis induced by NMU. Further studies are needed to elucidate the conditions of successful chemoprevention with Mel. PMID- 10813094 TI - Effects of retinyl acetate and melatonin on N-methyl-N-nitrosourea-induced mammary carcinogenesis in rats. A preliminary report. AB - The aim of this project was to evaluate the effect of retinyl acetate (RA), melatonin (Mel) and their combination on N-methyl-N-nitrosourea (NMU)-induced rat mammary carcinogenesis. Female Sprague-Dawley rats were given two intraperitoneal doses of NMU, each of 50 mg/kg of b.w. between 43rd to 57th postnatal day. The administration of RA started 11 days and the administration of Mel 12 days before the first dose of NMU. RA was given daily in a dose of 8.2 mg per animal and day at the base of the tongue. Mel was given as a solution (20 micrograms/ml of tap water) between 3 p.m. and 8 a.m., from 8 a.m. to 3 p.m. the animals were drinking tap water only. The experiment was finished 22 weeks after the first administration of the carcinogen. The tumour incidence in the control group was 88%, in the group treated with RA 80% and in the group treated with Mel 61%. A substantial decrease in tumour incidence to 37% was noted in the group treated with RA plus Mel. Significant differences in incidence were noted in the group treated with the combination of RA and Mel as compared to the control group and the group treated with RA. Chemoprevention lengthened the latency significantly in the group treated with Mel and with the combination of RA and Mel. The decrease in tumour frequency per group was confirmed in the group treated with the combination of RA and Mel; differences between groups in the frequency per tumour-bearing animal were not observed. The volume of mammary tumours in the groups treated with chemopreventive agents was not changed. PMID- 10813095 TI - Role of melanotransferrin (p97) in non-transferrin iron uptake by HeLa and K562 cells. AB - We tested whether melanotransferrin (p97), an iron-binding protein of the plasma membrane, is involved in the transport of non-transferrin iron into human HeLa and K562 cells. The expression of melanotransferrin was detected in HeLa but not in K562 cells. PI-PLC treatment dramatically decreased (to 20% of the original value) p97 expression by HeLa cells. However, the rate of iron uptake from 55Fe ferric citrate by PI-PLC-treated HeLa cells was comparable or only slightly lower (80-100%) than the rate of iron uptake by untreated cells. PI-PLC treatment had no effect on iron uptake by K562 cells. These findings strongly support the suggestion that melanotransferrin does not play a substantial role in non transferrin iron uptake by either HeLa or K562 cells. PMID- 10813096 TI - Preimplantation development of giant triploid zygotes in the mouse. AB - Giant oocytes or two-cell embryos have been reported in various mammalian species. They may arise during multiplication of oogonia, after fusion of two oogonia or, more probably, when nuclear division is not accompanied by cytoplasmic division. The ultimate fate of these giant embryos is not well known. In our laboratory, giant two-cell mouse embryos have been occasionally observed. Recently, we observed two giant one-cell zygotes in the same species. Both showed two female pronuclei and one male pronucleus, as well as two second polar bodies localized at opposite poles of the embryo. These two giant zygotes showed normal viability and developmental capacity. Their triploid nature was confirmed by cytogenetic analysis. In order to study this interesting phenomenon in more detail, we produced giant oocytes containing two germinal vesicles by cell fusion and cultured them in vitro. About one third of them extruded two first polar bodies; in the second group only one polar body was observed, whilst the last group was without polar bodies. When parthenogenetically activated, the consistent answer analogical to that observed in "in vivo" oocytes was only observed when oocytes with two polar bodies were activated. The implication for IVF technologies is discussed. PMID- 10813097 TI - New monoclonal antibodies specific for microtubule-associated protein MAP2. PMID- 10813098 TI - Monoclonal antibodies against type-A staphylococcal enterotoxin. PMID- 10813099 TI - Inhibition of gonadotropin surge by a brief mid-cycle regimen of ethinyl estradiol and norethindrone: possible role in in vitro fertilization. AB - Various methods to prevent premature luteinizing hormone (LH) surge and improve cycle control during hyperstimulation for in vitro fertilization (IVF) are standard of care. The purpose of the present study was to determine the influence of a 5-day regimen of ethinyl estradiol (EE) and norethindrone (NET) on folliculogenesis, gonadotropin surge, and ovulation. In a prospective randomized and comparative study, ten patients were assigned to two groups. A combination of 50 micrograms of EE and 1 mg of NET was used in groups I and II from days 6 through 10, and days 8 through 12, respectively. Blood samples and transvaginal ultrasound imaging were carried out throughout a 28-day cycle. Follicular diameter, plasma levels of LH, follicle-stimulating hormone (FSH), estradiol and progesterone, and endometrial thickness were determined. No LH surge or ovulation was detected in any patient studied. Peak estradiol concentrations were not significantly different between the groups (152.04 +/- 107.1 pg/ml vs 162.1 +/- 56.1 pg/ml [mean +/- SD] for groups I and II, respectively). No differences were noted between the groups for serum concentrations of FSH (range: 2-9 mIU/ml) or LH (range: 2-10 mIU/ml) for any given cycle day. Mean follicular diameters were not different between groups I and II (20.5 +/- 8.1 mm2 vs 20.6 +/- 14.2 mm2). Ultrasound assessment of mid-cycle follicular growth revealed diameters ranging from 18.5 mm2 to 34.0 mm2. Endometrial thickness ranged from 8 to 10 mm. There was no evidence of ovulation on ultrasound examination and either persistence or gradual resolution of follicles through the luteal phase. Peak serum concentrations at mid-luteal phase were < or = 2 ng/ml. In this pilot study, the combination of EE and NET restricted to a 5-day course beginning on day 6 or 8 permitted folliculogenesis but effectively inhibited midcycle LH surge and ovulation. Such regimens may have a role in IVF cycles for prevention of premature LH surges, especially as stimulation regimens evolve toward decreased gonadotropin use for stimulation and strict FSH preparations with the potential need for less complete pituitary suppression. PMID- 10813100 TI - A prospective and randomized study of ovarian stimulation for ICSI with recombinant FSH versus highly purified urinary FSH. AB - A prospective and randomized study of ovarian stimulation with human recombinant follicle-stimulating hormone (r-FSH; Gonal-F) versus highly purified urinary FSH (u-FSH-HP; Metrodin-HP) was conducted on patients submitted to an intracytoplasmic sperm injection (ICSI) program. A total of 120 patients aged 37 years or less were stimulated in a randomized manner with r-FSH (group I = 60 patients) or u-FSH-HP (group II = 60 patients). All received a fixed dose of FSH for 7 days and on the 8th day of stimulation the doses started to be adapted according to ovarian response. Human chorionic gonadotropin (hCG) at the dose of 5000 IU to 10,000 IU was administered to both groups when at least one follicle presented a diameter > or = 17 mm. The ovarian response did not differ significantly between groups I and II in terms of number of follicles > or = 16 mm (group I = 6.2 +/- 3.2; group II = 6.7 +/- 2.9; p = 0.26), number of oocytes collected (group I = 10.7 +/- 6.8; group II = 10.5 +/- 5.7; p = 0.91), number of oocytes in metaphase II (group I = 9.2 +/- 5.8; group II = 8.2 +/- 4.8; p = 0.56) or number of immature oocytes (group I = 1.8 +/- 0.9; group II = 1.9 +/- 1.7; p = 0.62). The normal fertilization rate after ICSI did not differ significantly between treatments (group I = 69.4 +/- 25; group II = 66.5 +/- 23; p = 0.38). No cases of cancellation of ovarian stimulation or of severe ovarian hyperstimulation syndrome occurred in either group. The total number of embryos obtained from patients who used r-FSH (group I = 6.3 +/- 4.5) was similar (p = 0.46) to the number obtained from patients who used u-FSH-HP (group II = 5.5 +/- 3.7), as also was the number of transferred embryos (group I = 2.8 +/- 0.8; group II = 2.6 +/- 0.9; p = 0.27). Implantation rate (26.1%) and clinical pregnancy rates per puncture (36.7%) and per embryo transfer (37.9%) were higher in patients who used r-FSH than in patients who used u-FSH-HP (19.5%, 31.7% and 32.2%, respectively), but the differences were not statistically significant. The abortion rate (p = 0.32) did not differ between groups (group I = 4.5%, n = 1 versus group II = 15.7%, n = 3). Thus far, the data do not demonstrate significant differences in ovary stimulation with r-FSH versus u-FSH in patients whose indication for assisted reproduction was the male factor. PMID- 10813101 TI - Gonadotropin stimulation following GnRH-a priming for poor responders in in vitro fertilization-embryo transfer programs. AB - The effect of gonadotropin-releasing hormone agonist (GnRH-a) administration before gonadotropin superovulation on the stimulation characteristics of poor responder patients was assessed in an in vitro fertilization (IVF) program. Thirty consecutive patients who had exhibited low ovarian response (fewer than four retrieved oocytes) in at least two previous IVF cycles (control cycles, n = 60), were eligible for the study. GnRH-a (nafarelin) was administered daily for 7 10 days from the mid-luteal phase of the previous cycle until the first day of menstruation. Menotropin treatment was commenced on cycle day 3 (with no additional GnRH-a) (study cycles, n = 39). A significantly higher number of oocytes was retrieved (p < 0.0002) and a higher number of embryos transferred (p < 0.003) in the study cycles than in the control cycles. No cases of premature luteinizing hormone surge were recorded. Pregnancy rates per embryo transfer and per cycle were 10.4% and 7.7% for the study cycles and 2.8% and 1.6% for the control cycles, respectively. GnRH-a, administered prior to gonadotropin treatment, should be an additional option of ovulation induction protocol for poor responders in IVF programs. PMID- 10813102 TI - A comparative study of the effects of two oral contraceptives containing dienogest or desogestrel on the human immune system. AB - The effects of two oral contraceptive combinations, dienogest 2.0 mg plus ethinyl estradiol 0.03 mg (Valette) and desogestrel 0.15 mg plus ethinyl estradiol 0.02 mg (Lovelle), on the human immune system were compared over one treatment cycle of 31 patients (n = 15 and n = 16, respectively). Lovelle but not Valette significantly increased the numbers of lymphocytes, monocytes and granulocytes. Valette decreased CD4 lymphocytes after 10 days' treatment; Lovelle had the opposite effect. Lovelle increased CD19 and CD23 after 21 days' treatment. Phagocytic activity was unaffected by either treatment. After 10 days' treatment, both contraceptives reduced serum IgA, IgG and IgM, which remained lowered at day 21 with Lovelle but returned to baseline with Valette. Secretory IgA was unaffected by either contraceptive. Neither treatment affected levels of interleukins, except for a significant difference between the treatment groups for interleukin-6 after 10 days' treatment that disappeared after 21 days' treatment. Levels of non-immunoglobulin serum components fluctuated; macroglobulin was increased with Valette. However, total protein and albumin levels were reduced more with Lovelle than with Valette. Complement factors also fluctuated. There was no evidence for any sustained immunosuppression with either Valette or Lovelle. PMID- 10813103 TI - Evaluation of matrix metalloproteinases-1 and -3 concentrations in the tunica albuginea, the apical wall of atretic follicles and the corpus luteum of normal human ovaries. AB - Matrix metalloproteinases-1 (MMP-1) and -3 (MMP-3) are proteolytic enzymes involved in remodeling the ovarian extracellular matrix throughout the menstrual cycle. The aim of the present study was to evaluate the tissue concentrations of MMP-1 and MMP-3 in the apical wall of atretic follicles (androstenedione/estradiol ratio > 4), tunica albuginea dissected from the ovarian surface overlying areas devoid of follicles, corpus luteum, and tunica albuginea covering the corpus luteum. After extraction of MMPs from the tissue samples, their concentrations in the extracts were measured by enzyme-linked immunosorbent assays (ELISA). Significantly less MMP-1 was detected in the apical wall of atretic follicles compared to tunica albuginea taken from sites devoid of follicles. These data indicate that atresia is associated with relatively low concentrations of MMP-1 in the apical wall of the follicle. Moreover, there was a negative correlation between the amount of MMP-3 and the diameter of follicle. These data suggest that both MMPs play an important role in the final step of atresia. The amount of MMP-1 in the corpus luteum was several times lower than in the other tissues. This is likely due to stabilization of the extracellular matrix during the period of the corpus luteum maintenance. The concentration of MMP-3 did not differ significantly among the examined tissues. PMID- 10813104 TI - Polycystic ovary syndrome: evidence for reduced 3 beta-hydroxysteroid dehydrogenase gene expression in human luteinizing granulosa cells. AB - In a preview study we found that luteinizing granulosa cells from follicles of patients with polycystic ovary syndrome (PCOS) have a reduced capacity to synthesize progesterone in vitro. Because the 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta-HSD) is an important enzyme for the biosynthesis of progesterone, the reduced capacity of PCO luteinizing granulosa cells to synthesize progesterone in vitro may be due to reduced 3 beta HSD gene expression. A reverse transcriptase polymerase chain reaction for 3 beta HSD was performed and the relative intensity of signals for 3 beta-HSD was evaluated using computer-assisted densitometry. Cells from polycystic ovaries expressed less 3 beta-HSD in follicles < or 10 mm (p < 0.05) and in follicles > or 16 mm (p < 0.05) than cells from normal ovaries. Furthermore, after human chorionic gonadotropin stimulus (50 ng/ml), cells from polycystic ovaries expressed less 3 beta-HSD in follicles > or = 16 mm (p < 0.01) than cells from normal ovaries. The data show that there is a specific change in the gene expression of 3 beta-HSD in PCO granulosa cells resulting in a suppressed capacity to secrete progesterone. PMID- 10813105 TI - Treatment of hirsutism with lowest-dose flutamide (62.5 mg/day). AB - Flutamide has been used successfully in the treatment of hirsutism. However, flutamide may cause important side-effects. The aim of this study was to evaluate the clinical and hormonal effects of lowest-dose flutamide therapy. Sixty-one women with hirsutism aged 18-37 years (mean +/- SD 23.4 +/- 5.9 years) were included in the study. Patients received 62.5 mg flutamide once per day for a period of 12 months. A hirsutism score was determined according to a modified Ferriman-Gallwey scoring system. Before therapy, multiscreen blood chemistry, hormonal analysis and complete blood counts were performed. These parameters and hirsutism scores were repeated at 3, 6, 9 and 12 months during therapy. The modified Ferriman-Gallwey scores significantly decreased from 19.1 +/- 4.9 to 5.8 +/- 3.3 during the study (p < 0.0001). The percentage reductions in hirsutism scores (mean +/- SD) were 60.3 +/- 14.4% at 6 months, and 70.3 +/- 13.2% at 12 months. No significant side-effects or modifications in the menstrual cycles were observed. There were no significant differences in any of the hormone levels during therapy. In conclusion, the lowest dose of flutamide, 62.5 mg/day, is a well-tolerated therapeutic agent and can be used in the treatment of hirsutism. PMID- 10813106 TI - Predictive value of serum androstenedione basal levels on the choice of gonadotropin or laparoscopic ovarian electrocautery as ovulation induction in clomiphene citrate-resistant patients with polycystic ovary syndrome. AB - Therapeutic approaches to chronic anovulation from polycystic ovaries in clomiphene-resistant infertile patients are under debate. This study discusses evidence that supports the possible predictive value of serum basal level of androstenedione in the choice of the better therapy between laparoscopic ovarian electrocautery and ovulation induction. Lower androstenedione levels seem to be correlated with a better ovarian response after ovulation induction with gonadotropins, while high androstenedione levels are associated with a higher incidence of conception after laparoscopic ovarian electrocautery. Obesity does not seem to represent a hindrance to laparoscopic treatment. PMID- 10813107 TI - Presence of a Gs alpha mutation in an adrenal tumor expressing LH/hCG receptors and clinically associated with Cushing's syndrome. AB - We describe the case of a patient with Cushing's syndrome due to a functioning adrenal adenoma. There was a pronounced increase in serum and urinary cortisol after administration of human chorionic gonadotropin. Immunocytochemistry revealed positive immunostaining for LH/hCG receptors. Molecular analysis documented the presence of a gsp mutation at codon 201 (CGT to TGT). The existence of this type of hCG-responsive adrenal tumor may help explain the higher prevalence of cortisol-secreting adrenal tumors versus pituitary-dependent disease in pregnant women with Cushing's syndrome as well as some reported cases of remission following delivery. PMID- 10813108 TI - Antiestrogens reduce plasma levels of endothelin-1 without affecting nitrate levels in breast cancer patients. AB - Tamoxifen protects against myocardial infarction through mechanisms that are poorly understood. We studied the effects of tamoxifen and another antiestrogen, toremifene, on the production of vasoconstrictive endothelin-1 and of vasodilatory nitric oxide in 44 postmenopausal patients with breast cancer. These started treatment, in randomized order, with either tamoxifen (20 mg/day; n = 25) or toremifene (40 mg/day; n = 19). Plasma samples collected before treatment and after 6 and 12 months of both regimens were assayed for endothelin-1 with a specific radioimmunoassay and for nitrite/nitrate with a method based on the Griess reaction. The antiestrogen group as a whole showed a fall in endothelin-1 at 6 months (5.9 +/- 3.3%; p = 0.06) (mean +/- SE) and at 12 months (7.1 +/- 5.5%; p = 0.03). This fall was solely due to toremifene, the use of which was associated with falls in endothelin-1 at 6 months (12.9 +/- 4.7%; p = 0.01) and 12 months (9.2 +/- 6.2%; p = 0.06). The antiestrogen regimen failed to affect plasma nitric oxide significantly but nevertheless the ratio between nitric oxide and endothelin-1 rose by 31.6 +/- 13.3% at 6 months and by 35.6 +/- 15.3% at 12 months in the antiestrogen users, an effect similar in the tamoxifen and toremifene groups. We conclude that antiestrogens may protect against myocardial infarction by preventing the release of endothelin-1 and by shifting the balance between nitric oxide and endothelin-1 to the dominance of the former. Our data predict that toremifene and tamoxifen at the doses studied here will provide similar cardiovascular protection. PMID- 10813109 TI - Association of vitamin D receptor gene polymorphism with bone mineral density in Slovenian postmenopausal women. AB - Osteoporosis is a common bone disease which affects one in three women after the 60th year of life and is a major cause of morbidity in older people. To identify patients with osteoporosis, measurement of bone mineral density (BMD) is recommended. The association of BMD with vitamin D receptor (VDR) genotype in Slovenian postmenopausal women was studied. We determined VDR genotype in 102 late postmenopausal women aged 47-77 years. BMD measurements were performed at the level of the lumbar spine (L2-L4) by dual X-ray absorptiometry. Our data show significantly lower BMD in BB women compared to those with bb genotype. The relative distribution of VDR genotypes and alleles in the Slovenian population was 18.6:57.8:23.6% for BB:Bb:bb, respectively. The results are consistent with those of a previous study which found an excellent correlation between BB VDR genotype and low BMD. The data were derived from a relatively small, but ethnically homogeneous population of the same socioeconomic status, with very similar dietary and physical activity habits. Dietary habits in particular seem to be important because of the relatively low calcium intake which may enhance the phenotypic expression of VDR gene polymorphisms. PMID- 10813110 TI - A case of primary amenorrhea, diabetes and anosmia. AB - This case details a patient with primary amenorrhea with an unusual cause. She presented at age 16 with short stature, minimal sexual development and no prior menses. Her history was significant for poorly controlled type 1 diabetes. She had been evaluated previously for growth hormone deficiency, and had received a short course of growth hormone therapy. Of greatest significance was the fact that she had also had a decreased sense of smell since her youth. Although a previous computerized tomography scan had been reported as normal, follow-up magnetic resonance imaging demonstrated the absence of olfactory bulbs. Smell testing confirmed the absence of smell and testing of gonadotropin releasing hormone demonstrated an inadequate response. All of these features suggested Kallmann syndrome. This syndrome commonly presents with delayed onset of puberty and decreased or absent sense of smell. There are also many associated features, and the disease is remarkable for its great genotypic and phenotypic variability. Current understanding of its pathogenesis, the commonly associated features of Kallmann syndrome and the impact of diabetes on growth and sexual development are reviewed. PMID- 10813111 TI - Beyond prediction to explanation in risk assessment research. A comparison of two explanatory theories of criminality and recidivism. PMID- 10813112 TI - Sexual offense severity and victim gender preference. A comparison of adolescent and adult sex offenders. PMID- 10813113 TI - Human rights of persons with mental disabilities. The European Convention of Human Rights. AB - It is not necessary to recount the numerous charters and declarations ... to understand human rights.... All persons are born free and equal in dignity and rights. Everyone ... is entitled to all the rights and freedoms set forth in the international human rights instruments without discrimination, such as the rights to life, liberty, security of the person, privacy, health, education, work, social security, and to marry and found a family. Yet, violations of human rights are a reality to be found in every corner of the globe. PMID- 10813114 TI - Neuropsychological and diagnostic differences between recidivistically violent not criminally responsible and mentally ill prisoners. PMID- 10813115 TI - The British Columbia Review Panel. Factors influencing decision-making. PMID- 10813116 TI - Laparoscopic surgery for the cure of colorectal cancer: results of a German five center study. AB - PURPOSE: The aim of this study was to assess the feasibility and safety of laparoscopic surgery for the cure of colorectal cancer with emphasis on oncologic follow-up in particular. METHODS: A study was performed of patients with colorectal cancer treated by laparoscopy in five German centers between May 1991 and September 1997. Surgical and pathologic data were recorded in an anonymous registry database and analyzed by type of resection. Standard procedures were sigmoid or left colectomy, anterior resection, abdominoperineal resection, and right hemicolectomy. Follow-up information included incidence of local, distant, and port site recurrence and cancer-related death. RESULTS: A total of 399 patients (212 females) with a mean age of 66.6 years underwent laparoscopic curative resections (sigmoid resection, 89; left colectomy, 11; anterior resection, 157; abdominoperineal resection, 102; right hemicolectomy, 40). Conversion was necessary in 6.3 percent (n = 25). Complications requiring reoperation occurred in 9 percent (n = 35). Complications that were treated conservatively occurred in 27.6 percent (n = 110). Thirty-day mortality was 1.8 percent (n = 7). First bowel movements resumed on the third postoperative day; patients did not use analgesics after a mean of five days. Mean postoperative hospitalization was two weeks. According to International Union Against Cancer classification, 147 patients had Stage I cancer, 35 had Stage II cancer, and 217 underwent curative resection for Stage III cancer. Mean number of lymph nodes resected was 12.1. At a mean follow-up of 30 months, one port site recurrence was documented. No local recurrence was observed after curative resection of Stage I colorectal cancer. Of 399 patients, local recurrence occurred in 6 patients (Stage II, 2; Stage III, 4), and distant metastases were documented in 25 patients (Stage I, 3; Stage II, 3; Stage III, 19). The highest incidence of cancer-related death occurred after abdominoperineal resection (4.9 percent). CONCLUSION: To assess the role of laparoscopic colorectal surgery for the cure of cancer objectively, prospective randomized trials are necessary. PMID- 10813117 TI - Fecal Incontinence Quality of Life Scale: quality of life instrument for patients with fecal incontinence. AB - PURPOSE: This goal of this research was to develop and evaluate the psychometrics of a health-related quality of life scale developed to address issues related specifically to fecal incontinence, the Fecal Incontinence Quality of Life Scale. METHODS: The Fecal Incontinence Quality of Life Scale is composed of a total of 29 items; these items form four scales: Lifestyle (10 items), Coping/Behavior (9 items), Depression/Self-Perception (7 items), and Embarrassment (3 items). RESULTS: Psychometric evaluation of these scales demonstrates that they are both reliable and valid. Each of the scales demonstrate stability over time (test/retest reliability) and have acceptable internal reliability (Cronbach alpha >0.70). Validity was assessed using discriminate and convergent techniques. Each of the four scales of the Fecal Incontinence Quality of Life Scale was capable of discriminating between patients with fecal incontinence and patients with other gastrointestinal problems. To evaluate convergent validity, the correlation of the scales in the Fecal Incontinence Quality of Life Scale with selected subscales in the SF-36 was analyzed. The scales in the Fecal Incontinence Quality of Life Scale demonstrated significant correlations with the subscales in the SF-36. CONCLUSIONS: The psychometric evaluation of the Fecal Incontinence Quality of Life Scale showed that this fecal incontinence-specific quality of life measure produces both reliable and valid measurement. PMID- 10813118 TI - Prospective assessment of primary rectal cancer response to preoperative radiation and chemotherapy using 18-fluorodeoxyglucose positron emission tomography. AB - PURPOSE: The purpose of this prospective study was to determine the ability of fluorine-18 fluorodeoxyglucose positron emission tomography to assess extent of pathologically confirmed rectal cancer response to preoperative radiation and 5 fluorouracil-based chemotherapy. METHODS: Patients with primary rectal cancer deemed eligible for preoperative radiation and 5-fluorouracil-based chemotherapy because of a clinically bulky or tethered tumor or endorectal ultrasound evidence of T3 and/or N1 were prospectively enrolled. Positron emission tomography and CT scans were obtained before preoperative radiation and 5-fluorouracil-based chemotherapy (5,040 cGy to the pelvis and 2 cycles of bolus 5-fluorouracil with leucovorin) and repeated four to five weeks after completion of radiation and 5 fluorouracil-based chemotherapy. In addition to routine pathologic staging, detailed assessment of rectal cancer response to preoperative radiation and 5 fluorouracil-based chemotherapy was performed independently by two pathologists. Positron emission tomography parameters studied included conventional measures such as standardized uptake value (average and maximum), positron emission tomography-derived tumor volume (size), and two novel parameters: visual response score and change in total lesion glycolysis. RESULTS: Of 21 patients enrolled, prospective data (pretreatment and posttreatment positron emission tomography, and complete pathologic assessment) were available on 15 patients. All 15 demonstrated pathologic response to preoperative radiation and 5-fluorouracil based chemotherapy. This was confirmed in 100 percent of the cases by positron emission tomography compared with 78 percent (7/9) by CT. In addition, one positron emission tomography parameter (visual response score) accurately estimated the extent of pathologic response in 60 percent (9/15) of cases compared with 22 percent (2/9) of cases with CT. CONCLUSIONS: This pilot study demonstrates that fluorine-18 fluorodeoxyglucose positron emission tomography imaging adds incremental information to the preoperative assessment of patients with rectal cancer. However, further studies in a larger series of patients are needed to verify these findings and to determine the value of fluorine-18 fluorodeoxyglucose positron emission tomography in a preoperative strategy aimed at identifying patients suitable for sphincter-preserving rectal cancer surgery. PMID- 10813119 TI - Laparostomy for severe intra-abdominal infection complicating colorectal disease. AB - PURPOSE: The aim of this study was to evaluate the use of laparostomy in the management of patients with severe intra-abdominal infection resulting from colorectal disease. METHODS: Seven patients, four with inflammatory bowel disease, two with colorectal carcinoma, and one with diverticular perforation, underwent laparostomy during a six-year period for postoperative, severe, intra abdominal infection. RESULTS: The median age was 42 years, the mean Acute Physiology and Chronic Health Evaluation II score was 22.7, and the observed mortality was 28.6 percent (2/7 patients). In one patient the laparostomy was closed at 11 days; in all the others the wound was left to heal by granulation and contraction, and two of these later required reconstructive surgery. The median follow-up was three years and seven months. CONCLUSION: Laparostomy is an effective and practical method of managing patients with severe intra-abdominal infection as a result of colorectal disease. PMID- 10813120 TI - Closed vs. open hemorrhoidectomy--is there any difference? AB - PURPOSE: The aim of this study was to compare closed (Ferguson) hemorrhoidectomy to open (Milligan-Morgan) hemorrhoidectomy regarding postoperative conditions, complications, and long-term results. METHOD: This was a randomized study of 77 patients with second-degree or third-degree hemorrhoids suitable for hemorrhoidectomy. In 39 patients the Milligan-Morgan procedure was used, and in 38 patients the Ferguson procedure was used. Details of operations, postoperative complications, and length of postoperative stay were recorded. Pain was assessed from a visual analog scale and by registration of postoperative analgesic medication. Follow-up was done at three weeks, six weeks, and by visit or telephone interview after at least a year. RESULTS: No statistically significant differences were found between the two methods regarding complications, pain, or postoperative stay. There were four reoperations for bleeding, all after Milligan Morgan operations. At follow-up after three weeks 86 percent of the Ferguson patients had completely healed wounds, and none had signs of infection. Of the Milligan-Morgan patients, only 18 percent had completely healed wounds, and symptoms of delayed wound healing were significantly more frequent. One patient had a superficial wound infection. After one year more than 10 percent in each group had recurrent hemorrhoids with symptoms. CONCLUSION: Both methods are fairly efficient treatment for hemorrhoids, without serious drawbacks. The closed method has no advantage in postoperative pain reduction, but wounds heal faster, and the risk of wound dehiscence seems exaggerated. PMID- 10813121 TI - Long-term results and functional outcome after Ripstein rectopexy. AB - PURPOSE: The aim of this study was to evaluate operative mortality, morbidity, and functional results after Ripstein rectopexy for rectal prolapse and internal rectal intussusception. METHODS: Sixty-nine patients with rectal prolapse and 43 with internal rectal intussusception were included. All patient records were studied and complications registered. Long-term follow-up was possible in 105 patients and performed by clinical examination and standardized interview, telephone interview, or patient records. Seventy-six patients were prospectively evaluated, comparing bowel function before and after rectopexy. RESULTS: There was no operative mortality. Operative morbidity was 33 percent, and most complications were minor. Severe early complications included one large-bowel obstruction and one transient ureteric stenosis. Median time of follow-up was seven years in patients with rectal prolapse and 5.4 years in patients with internal rectal intussusception. Late complications included two rectovaginal fistulas and one lethal sigmoid fecaloma. Five patients underwent subtotal colectomy for severe constipation. There was one recurrent prolapse (1.6 percent). Functional evaluation showed that incontinence improved (P = 0.049), whereas the number of bowel movements per week decreased (P < 0.001). Frequency of emptying difficulties did not change significantly in patients with rectal prolapse but increased in patients with internal rectal intussusception (P = 0.038). CONCLUSION: Ripstein rectopexy can be performed with low mortality and recurrence rate, but with a high early complication rate. There were also some serious late complications. Continence was improved, although increased constipation was a problem in some patients, especially among those with internal rectal intussusception. PMID- 10813122 TI - Prospective, blinded comparison of laparoscopic ultrasonography vs. contrast enhanced computerized tomography for liver assessment in patients undergoing colorectal carcinoma surgery. AB - PURPOSE: To prospectively and blindly compare intraoperative laparoscopic ultrasonography to preoperative contrast-enhanced computerized tomography in detecting liver lesions in colorectal cancer patients. Additionally, we compared conventional (open) intraoperative ultrasonography with bimanual liver palpation to contrast-enhanced computerized tomography in a subset of patients. METHODS: From December 1995 to March 1998, 77 consecutive patients underwent curative (n = 63) or palliative (n = 14) resections for colorectal cancer. All patients undergoing curative resections were randomized to either laparoscopic (n = 34) or conventional (n = 29) surgery after informed consent. All patients underwent contrast-enhanced computerized tomography, diagnostic laparoscopy, and laparoscopic ultrasonography before resection. In those patients who had conventional procedures, intraoperative ultrasonography with bimanual liver palpation was also done. All laparoscopic ultrasonography and intraoperative ultrasonography evaluations were performed by one of two radiologists who were blinded to the CT results. All hepatic segments were scanned using a standardized method. The yield of each modality was calculated using the number of lesions identified by each imaging modality divided by the total number of lesions identified. RESULTS: In 43 of the 77 patients, both the laparoscopic ultrasonography and CT scan were negative for any liver lesions. In 34 patients, a total of 130 lesions were detected by laparoscopic ultrasonography, CT, or both. When compared with laparoscopic ultrasonography, intraoperative ultrasonography with bimanual liver palpation identified one additional metastatic lesion and no additional benign lesions. laparoscopic ultrasonography identified two patients with mets who had negative preoperative contrast-enhanced computerized tomography. CONCLUSIONS: Laparoscopic ultrasonography of the liver at the time of primary resection of colorectal cancer yields more lesions than preoperative contrast-enhanced computerized tomography and should be considered for routine use during laparoscopic oncologic colorectal surgery. PMID- 10813123 TI - Cecostomy is a useful surgical procedure: study of 113 colonic obstructions caused by cancer. AB - PURPOSE: There is a large choice of treatment for obstructing carcinoma of the left colon. We report our experience of tube cecostomy as the initial treatment for obstructing colonic carcinoma followed by elective resection. METHODS: From 1975 to 1995, 113 patients presenting with colonic obstruction caused by cancer were initially treated by tube cecostomy. RESULTS: The cecostomy was performed under local anesthesia in 26 cases (23 percent) and general anesthesia in 87 cases (77 percent). In the postoperative period 15 patients died (13 percent) and 26 (23 percent) had wound infection in the area around the cecostomy. A second operation performed on the 98 surviving patients comprised 74 left colonic resections with anastomosis, 9 without anastomosis (Hartmann's operation), 1 right colectomy, 3 total colectomies eliminating the cecostomy, 3 internal bypasses, and 8 proximal lateral colostomies. Surgical closure of the cecostomy was performed during six of the second operations. No deaths occurred from any of the second operations. The cecostomy closed spontaneously in 78 patients (89 percent). In ten cases (11.4 percent) a third operation was performed to close the cecostomy, without mortality. CONCLUSIONS: Comparison our cecostomy results with published studies of proximal diverting loop colostomies for the same indications showed comparable mortality after the first operation. Cecostomy decrease mortality of the second operation. This retrospective study suggests that cecostomy is a useful and less invasive surgical procedure for patients presenting with colonic obstruction caused by cancer. PMID- 10813124 TI - Improving the reinforcement of parastomal tissues with Marlex mesh: laboratory study identifying solutions to stomal aperture distortion. AB - PURPOSE: Parastomal hernia formation commonly complicates permanent stomas and represents a significant and frequently recurrent management problem, regardless of the method of repair. Prosthetic material reinforcement of parastomal tissues offers the best results. However, problems with unravelling of mesh fibers along cut margins leading to aperture enlargement and hernia recurrence may occur. Raised intra-abdominal pressure in the early postoperative period before incorporation of the mesh into surrounding tissues may result in hernia formation if the aperture size in the mesh increases. METHODS: Assessment of the physical properties of Marlex mesh was performed in a materials testing laboratory, using standardized tests to simulate the stresses imposed on in situ mesh. RESULTS: Holes cut in Marlex mesh were found to enlarge and distort at loads simulating intra-abdominal pressure changes. Reinforcement with a polypropylene pursestring suture was found to stabilize the periaperture mesh fibers and maintain the original area throughout tensions at least double maximal intra-abdominal pressures. Distensibility of intact sheets of mesh was found to vary by up to 100 percent, depending on the direction of the applied tension, and thus, mesh orientation in hernia repair has major implications. CONCLUSION: We propose that if mesh is used to reinforce abdominal wall tissues and is cut or fashioned to size, then the cut margins must be reinforced if the intended dimensions and functional integrity of the mesh are to be maintained. PMID- 10813125 TI - Laparoscopic colectomy compares favorably with colectomy by laparotomy for reduction of postoperative ileus. AB - PURPOSE: The aim of this study was to compare the length of postoperative ileus in patients undergoing colectomy by either laparotomy or laparoscopy. METHODS: A total of 166 patients were studied. These patients were divided into two groups: Group 1, in which colectomy was done laparoscopically, and Group 2, consisting of patients undergoing laparotomy. Both groups contained 83 patients who were matched for disease severity, indications for surgery, and procedure. Indications for surgery included sigmoid diverticulitis in 12 (14 percent) patients, polyps in 22 (27 percent), Crohn's disease in 21 (25 percent), colorectal cancer in 11 (13 percent), stoma reversal in 8 (10 percent), rectal prolapse in 3 (4 percent), and other indications in 6 (7 percent) in each group. Operations were colectomy with anastomosis (42 ileocolic, 26 colorectal, 6 colocolic, 4 ileorectal, and 2 ileal J pouch) or without anastomosis (3 abdominoperineal resections) performed by the same surgeons during the same time period January 1993 to October 1996). The nasogastric tube was removed from all patients immediately after surgery in both groups. All patients received a clear liquid diet on the first postoperative day, followed by a regular diet as tolerated. The nasogastric tube was reinserted if two or more episodes of emesis of more than 200 ml occurred in the absence of bowel movement. Patients were discharged from the hospital when tolerating a regular diet without evidence of ileus. Statistical analysis was performed using unpaired t-test and Fisher's exact probability test. RESULTS: The male-to-female ratio was 38 to 45 in both groups. A total of 10 (12 percent) and 23 (28 percent) patients in Group 1 and Group 2 had emesis (P = 0.02), and the rate of nasogastric tube reinsertion was 5 (6 percent) and 13 (16 percent), respectively (P > 0.05). There were significant differences between Groups 1 and 2 relative to the lengths of ileus (3.5 +/- 1.3 vs. 5.4 +/- 1.7 days, respectively; P < 0.001), hospitalization (6.6 +/- 3.3 vs. 8.1 +/- 2.5 days, respectively; P < 0.002), and operative time (170 +/- 60 vs. 114 +/- 46 minutes, respectively; P < 0.001). The morbidity rate was 16 (19.2 percent) and 18 (21.6 percent) in the laparoscopy and laparotomy groups, respectively. CONCLUSIONS: Although early oral intake is safe and can be tolerated by 84 percent of patients after colectomy by laparotomy, laparoscopic colectomy reduced the lengths of both postoperative ileus and hospitalization. PMID- 10813126 TI - Micronized purified flavonidic fraction compared favorably with rubber band ligation and fiber alone in the management of bleeding hemorrhoids: randomized controlled trial. AB - PURPOSE: The aim of this study was to assess the role of micronized purified flavonidic fraction in the management of bleeding nonprolapsed hemorrhoids. METHODS: Patients were randomly assigned to receive ispaghula husk alone, rubber band ligation plus ispaghula husk, or micronized purified flavonidic fraction plus ispaghula husk. Other colorectal diseases were excluded by colonoscopy. Blinded observers noted the time for bleeding to stop completely, recurrences, and treatment complications. RESULTS: A total of 162 patients were randomly assigned with no significant differences in the age and gender distributions among the groups. Hemorrhoidal bleeding was relieved most expediently in the micronized purified flavonidic fraction plus ispaghula husk group (ispaghula husk alone n = 66, mean (standard error of the mean) 10.6 (2.3) days; rubber band ligation plus ispaghula husk n = 57, 5.6 (1.1) days; micronized purified flavonidic fraction plus ispaghula husk n = 39, 3.9 (1.2) days; P = 0.03). However, there were no significant differences in the recurrences at six months of follow-up (ispaghula husk alone n = 8 (12 percent); rubber band ligation plus ispaghula husk n = 12 (21 percent); micronized purified flavonidic fraction plus ispaghula husk n = 2 (5.1 percent); P = 0.075). No complications or side-effects were noted. CONCLUSIONS: micronized purified flavonidic fraction used with fiber supplements rapidly and safely relieved bleeding from nonprolapsed hemorrhoids. PMID- 10813127 TI - ras p21 Isoprenylation inhibition induces flat colon tumors in Wistar rats. AB - PURPOSE: The effect of pravastatin, an inhibitor of ras p21 isoprenylation, on the gross type of colon tumors induced by azoxymethane was investigated in Wistar rats. METHODS: Rats received ten weekly subcutaneous injections of 7.4 mg/kg body weight of azoxymethane and intraperitoneal injections of 10 or 20 mg/kg body weight of pravastatin every other day until the end of the experiment at Week 45. RESULTS: Administration of pravastatin at both dosages had no significant effect on the incidence of colon tumors but significantly increased the incidence of rats with adenomas only. In contrast to the elevated adenomas in control rats, flat adenomas were significantly more prevalent in rats given pravastatin. Pravastatin at both doses significantly decreased the labeling index, but not the apoptotic index, of elevated adenomas, whereas it significantly decreased the labeling index but increased the apoptotic index of flat adenomas. Administration of pravastatin at both dosages also significantly decreased the amounts of membrane-associated ras p21 in colon tumors. CONCLUSIONS: These findings suggest that the ras oncogene may be closely related to the development of adenocarcinomas from adenomas and the development of elevated or polypoid tumors of the colon. PMID- 10813128 TI - Altered microperfusion at the rectal stump is predictive for rectal anastomotic leak. AB - PURPOSE: The aim of this study was to evaluate the reliability of intraoperative laser-Doppler measurements in predicting the occurrence of anastomotic leak in patients with colorectal cancer undergoing stapled straight anastomosis to the rectum. METHODS: A prospective study was undertaken on 55 patients with rectal cancer or distal sigmoid cancer programmed for elective curative surgery. In all patients transmural colonic blood flow was measured by laser-Doppler flowmetry technique before bowel manipulation (baseline measurement) and after vascular ligation and division. Comorbidities at admission, intraoperative events, associated surgical procedures, and clinical outcome were tested for any association with anastomotic leak. RESULTS: Postoperative mortality was 1.8 percent (1/55 patients), and the overall morbidity was 21.3 percent. Anastomotic leak occurred in eight patients (14.5 percent). After colonic division a blood flow reduction at the rectal stump was observed in 42 patients (76.3 percent) as compared with baseline measurement. The mean rectal stump flow reduction was 6.2 percent in patients without anastomotic leak, whereas in patients who developed anastomosis breakdown it was 16 percent (P < 0.001). Mean proximal stump flow reduction was 5.1 percent in the uncomplicated patients, whereas in patients who had an anastomosis breakdown it was 12.9 percent (P < 0.01). A positive linear correlation was found between decrease in blood flow and rate of anastomotic leak. CONCLUSION: Blood flow reduction at the rectal stump is associated with an increased risk of anastomotic leak. PMID- 10813129 TI - Risk factors for morbidity and mortality after colectomy for colon cancer. AB - PURPOSE: Comorbid conditions affect the risk of adverse outcomes after surgery, but the magnitude of risk has not previously been quantified using multivariate statistical methods and prospectively collected data. Identifying factors that predict results of surgical procedures would be valuable in assessing the quality of surgical care. This study was performed to define risk factors that predict adverse events after colectomy for cancer in Department of Veterans Affairs Medical Centers. METHODS: The National Veterans Affairs Surgical Quality Improvement Program contains prospectively collected and extensively validated data on more than 415,000 surgical operations. All patients undergoing colectomy for colon cancer from 1991 to 1995 who were registered in the National Veterans Affairs Surgical Quality Improvement Program database were selected for study. Independent variables examined included 68 preoperative and 12 intraoperative clinical risk factors; dependent variables were 21 specific adverse outcomes. Stepwise logistic regression analysis was used to construct models predicting the 30-day mortality rate and 30-day morbidity rates for each of the ten most frequent complications. RESULTS: A total of 5,853 patients were identified; 4,711 (80 percent) underwent resection and primary anastomosis. One or more complications were observed in 1,639 of 5,853 (28 percent) patients. Prolonged ileus (439/5,853; 7.5 percent), pneumonia (364/5,853; 6.2 percent), failure to wean from the ventilator (334/5,853; 5.7 percent), and urinary tract infection (292/5,853; 5 percent) were the most frequent complications. The 30-day mortality rate was 5.7 percent (335/5,853). For most complications, 30-day in-hospital mortality rates were significantly higher for patients with a complication than for those without. Thirty-day mortality rates exceeded 50 percent if postoperative coma, cardiac arrest, a pre-existing vascular graft prosthesis that failed after colectomy, renal failure, pulmonary embolism, or progressive renal insufficiency occurred. Preoperative factors that predicted a high risk of 30-day mortality included ascites, serum sodium >145 mg/dl, "do not resuscitate" status before surgery, American Society of Anesthesiologists classes III and IV OR V, and low serum albumin. CONCLUSIONS: Mortality rates after colectomy in Veterans Affairs hospitals are comparable with those reported in other large studies. Ascites, hypernatremia, do not resuscitate status before surgery, and American Society of Anesthesiologists classes III and IV OR V were strongly predictive of perioperative death. Clinical trials to decrease the complication rate after colectomy for colon cancer should focus on these risk factors. PMID- 10813130 TI - Prognostic value of microscopic peritoneal dissemination: comparison between colon and gastric cancer. AB - PURPOSE: We evaluated the incidence and prognostic relevance of microscopic intraperitoneal tumor cell dissemination of colon cancer in comparison with dissemination of gastric cancer as a rational for additive intraperitoneal therapy. METHODS: Peritoneal washouts of 90 patients with colon and 111 patients with gastric cancer were investigated prospectively. Sixty patients with benign diseases and 8 patients with histologically proven gross visible peritoneal carcinomatosis served as controls. Intraoperatively, 100 ml of warm NaCl 0.9 percent were instilled and 20 ml were reaspirated. In all patients hematoxylin and eosin staining (conventional cytology) was performed. Additionally, in 36 patients with colon cancer and 47 patients with gastric cancer, immunostaining with the HEA-125 antibody (immunocytology) was prepared. The results of cytology were assessed for an association with TNM category and cancer grade, based on all patients, and with patient survival, among the R0 resected patients. RESULTS: In conventional cytology 35.5 percent (32/90) of patients with colon cancer and 42.3 percent (47/111) of patients with gastric cancer had a positive cytology. In immunocytology 47.2 percent (17/36) of patients with colon cancer and 46.8 percent (22/47) of patients with gastric cancer were positive. In colon cancer, positive conventional cytology was associated with pT and M category (P = 0.044 and P = 0.0002), whereas immunocytology was only associated with M category (P = 0.007). No association was found between nodal status and immunocytology in colon cancer and with the grading. There was a statistically significant correlation between pT M category and conventional and immunocytology in gastric cancer (P < 0.0015/P = 0.007 and P < 0.001/P = 0.009, respectively). Positive immunocytology was additionally associated with pN category (P = 0.05). In a univariate analysis of R0 resected patients (no residual tumor), positive immunocytology was significantly related to an unfavorable prognosis in patients with gastric cancer only (n = 30). Mean survival time was significantly increased in patients with gastric cancer with negative cytology compared with positive cytology (1,205 (standard error of the mean, 91) vs. 771 (standard error of the mean, 147) days; P = 0.007) but not in patients with colon cancer (1,215 (standard error of the mean, 95) vs. 1,346 (standard error of the mean, 106) days; P = 0.55). CONCLUSIONS: Because microscopic peritoneal dissemination influences survival time after R0 resections only in patients with gastric but not with colon cancer, our results may provide a basis for a decision on additive, prophylactic (intraperitoneal) therapy in gastric but not colon cancer. PMID- 10813131 TI - Development of invasive adenocarcinoma in a long-standing diverted ileal J-pouch for ulcerative colitis: report of a case. AB - We report a case of a 50-year-old male with ulcerative colitis who developed well differentiated adenocarcinoma in the ileal J-pouch, which had been defunctioning for 18 years. The extension of the carcinoma in the pouch suggested that it had recently appeared in the pouch. Monitoring by endoscopic examination and biopsy or pouch excision seems to be an appropriate action if a pouch is out of the fecal stream. PMID- 10813132 TI - Hepatoid adenocarcinoma of the rectum arising in ulcerative colitis: report of a case. AB - We report a case of intestinal hepatoid adenocarcinoma, confirmed by albumin m RNA in situ hybridization, with subsequent metastatic spread to the liver in a male with a long-standing history of ulcerative colitis. This novel finding strongly suggests that ulcerative colitis can lead not only to conventional adenocarcinomas but also to hepatoid adenocarcinoma and highlights the mimicry of hepatocellular carcinoma by metastatic hepatoid adenocarcinoma liver nodules. PMID- 10813133 TI - Solitary neurofibroma of the anal canal: report of two cases. AB - Neurofibromas are benign nerve sheath tumors commonly found in patients afflicted with neurofibromatosis-1. In the absence of multiple neurofibromas or a diagnosis of neurofibromatosis-1, neurofibromas are referred to as solitary and have been reported to involve the skin, subcutaneous tissue, deep soft tissue, and viscera of almost all areas of the body. A neurofibroma involving the anal canal is a rare entity, with only one report in the literature. We present two cases in which large masses involving the anal canal of elderly females were locally resected and proved to be neurofibromas by histopathology. Although rare, these tumors should be considered in the differential diagnosis of patients presenting with an anal mass, because resection alone is the treatment of choice. PMID- 10813134 TI - Is it really not familial adenomatous polyposis? PMID- 10813135 TI - Cytomegalovirus infection as a cause of ileoanal pouchitis. PMID- 10813136 TI - Enterolith ileus: is there a role for nonoperative management? PMID- 10813137 TI - Treatment of enterocele by obliteration of the pelvic inlet. PMID- 10813138 TI - EMG of the external anal sphincter: needle is superior to surface electrode. PMID- 10813139 TI - Staplers and hemorrhoids. PMID- 10813140 TI - Conversion of ileorectal anastomosis. PMID- 10813141 TI - Molecular epidemiology of an outbreak caused by Salmonella enterica serovar Newport in Finland and the United Kingdom. AB - Between December 1997 and January 1998 an increase in the number of isolates of Salmonella enterica serovar Newport, a serotype rarely causing indigenous infections in Finland, was detected. This included two clusters of gastroenteritis following funeral meals. An inquiry via Enter-net revealed a concomitant increase in cases of S. Newport in the United Kingdom. To investigate the Finnish outbreak, a total of 56 S. Newport strains (22 from the outbreak period, 27 from pre- and post-outbreak period, and 7 from imported food producing animals) were studied by pulsed-field gel electrophoresis (PFGE); selected isolates were also phage typed. Two retrospective questionnaire studies evaluating food exposures among the funeral attendants were conducted. All isolates from the clusters had an identical PFGE pattern which was also found in 13 infections temporally close to but not associated with the clusters. The Finnish outbreak was caused by the same phage type as the one in the United Kingdom. In both clusters, an epidemiological link between illness and exposure to cured ham was found. In conclusion, the outbreak was not limited to the two clusters but was more widely spread both in and outside Finland. Early alarm systems of food-borne outbreaks and collaboration between European countries are needed for investigating international outbreaks. PMID- 10813142 TI - Multiresistant Salmonella Typhimurium DT104 infections of humans and domestic animals in the Pacific Northwest of the United States. AB - Salmonella Typhimurium definitive type 104 with chromosomally encoded resistance to five or more antimicrobial drugs (R-type ACSSuT+) has been reported increasingly frequently as the cause of human and animal salmonellosis since 1990. Among animal isolates from the northwestern United States (NWUS), R-type ACSSuT+ Typhimurium isolates increased through the early 1990s to comprise 73% of Typhimurium isolates by 1995, but subsequently decreased to comprise only 30% of isolates during 1998. NWUS S. Typhimurium R-type ACSSuT+ were consistently (99%) phage typed as DT104 or the closely related DTu302. S. Typhimurium isolates from cattle with primary salmonellosis, randomly selected from a national repository, from NWUS were more likely to exhibit R-type ACSSuT+ (19/24, 79%) compared to isolates from other quadrants (17/71, 24%; P < 0.01). Human patients infected with R-type ACSSuT+ resided in postal zip code polygons of above average cattle farm density (P < 0.05), while patients infected with other R-types showed no similar tendency. Furthermore, humans infected with R-type ACSSuT+ Typhimurium were more likely to report direct contact with livestock (P < 0.01) than humans infected with other R-types. PMID- 10813143 TI - Ecological study of Vibrio cholerae in Vellore. AB - Vellore is endemic for cholera due to Vibrio cholerae O1 and O139. In a previous study the prevalence of Vibrio cholerae in drinking water, lakes and sewage outfalls in a single 2-months period in Vellore, India was determined. In addition water samples from three sites were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. This follow on study has examined how the environmental distribution of V. cholerae at the same sites alters over a 12-month period and the relationship to the clinical pattern of cholera in Vellore. Samples of water were collected from fixed sites at three water bodies each month between April 1997 and March 1998. Bacteria isolated from samples were identified by standard biochemical tests and isolated strains of V. cholerae tested for their ability to agglutinate O1 and O139 antisera. Samples were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. The clinical isolation rate of V. cholerae in Vellore, maximum temperature and rainfall were also studied. The results demonstrate the presence in the environment of viable but non-cultivable (VNC) V. cholerae in 10 of 12 months of the study year as well as their viability. Their prevalence in the environment also correlated with the isolation of these pathogens from clinical samples over the same study period. PMID- 10813144 TI - A one year study of Escherichia coli O157 in raw beef and lamb products. AB - Between April 1996 and March 1997 we examined 5093 samples of raw beef and lamb products for the presence of E. coli O157. Samples were purchased from 81 small butchers' shops in south Yorkshire. In March 1997 we also examined five samples of dried mint for the presence of E. coli O157. Strains of E. coli O157 were isolated by enrichment culture in modified buffered peptone water followed by immunomagnetic separation and culture of magnetic beads onto cefixime tellurite sorbitol MacConkey agar. Strains were characterized by phage typing, toxin genotyping and plasmid analysis. Strains of E. coli O157 were isolated from 72 (1.4%) of 5093 samples; it was isolated from 36 (1.1%) of 3216 samples of beef products and from 29 (2.9%) samples of lamb products. The highest prevalence was found in lamb sausages and lamb burgers where E. coli O157 was isolated from 3 (4.1%) of 73 and 18 (3.7%) of 484 samples respectively. Strains of E. coli O157 were isolated most frequently during early summer. Strains of E. coli O157 were also isolated from 2 of 5 samples of dried mint although we did not determine how the mint had become contaminated. All isolates of E. coli O157 were Verocytotoxin producing as determined by both Vero cell assay and DNA hybridization for the genes encoding Verocytotoxin and all were positive for the eaeA gene. A combination of phage typing, toxin genotyping and plasmid profile subdivided the 72 strains of E. coli isolated into 20 different subtypes, of which 18 were indistinguishable from strains isolated previously from cattle and sheep; of these 18 strains, 8 were indistinguishable from strains isolated from human cases of infection during the study period. PMID- 10813146 TI - Incidence and prognosis of tuberculosis in patients with cirrhosis of the liver. A Danish nationwide population based study. AB - We examined the incidence rate and prognosis of tuberculosis in a cohort of patients with liver cirrhosis in Denmark. In a study cohort of 22675 patients with liver cirrhosis, we identified 151 cases of tuberculosis from 1977 to 1993. The incidence rate was 168.6 per 100000 person-years of risk, and the highest incidence rate was among men above 65 years of age, with an incidence rate of 246.0 per 100000 person-years of risk. The 30-day case-fatality rate was 27.3% and the 1-year case fatality rate was 47.7%. The results demonstrate that patients with liver cirrhosis are at increased risk of tuberculosis. Additionally, it is suggested that liver cirrhosis is an independent risk factor for tuberculosis, and that patients with liver cirrhosis who acquire tuberculosis have a poor prognosis. PMID- 10813145 TI - Haemolytic uraemic syndrome and Shiga toxin-producing Escherichia coli infection in children in France. The Societe de Nephrologie Pediatrique. AB - We conducted a study to determine the incidence of haemolytic uraemic syndrome (HUS) in children in France and to assess the role of Shiga-toxin-producing Escherichia coli (STEC) infection in the aetiology of HUS. In collaboration with the Societe de Nephrologie Pediatrique we undertook a retrospective review of all cases of HUS hospitalized from January 1993 to March 1995 and a 1-year prospective study (April 1995-March 1996) of epidemiological and microbiological features of cases of HUS. The polymerase chain reaction (PCR) procedure was used to detect stx, eae, e-hlyA genes directly from case stool samples. Serum samples from cases were examined for antibodies to lipopolysaccharide (LPS) of 26 major STEC serogroups. Two hundred and eighty-six cases were reported. The average incidence per year was 0.7/10(5) children < 15 years and 1.8/10(5) children < 5 years. During the prospective study, 122/130 cases were examined for evidence of STEC infection using PCR and/or serological assays and 105 (86%) had evidence of STEC infection. Serum antibodies to E. coli O157 LPS were detected in 79 (67%) cases tested. In conclusion, this study showed that STEC infection is an important cause of HUS in children in France, with a high proportion related to the O157 serogroup. PMID- 10813147 TI - Molecular characterization of pncA gene mutations in Mycobacterium tuberculosis clinical isolates from China. AB - A sample of 35 pyrazinamide (PZA)-resistant and 30 PZA-susceptible clinical isolates recovered from Beijing and Taiyuan City, China were characterized by SSCP and sequence analysis for mutations in the pncA gene that encodes the Mycobacterium tuberculosis PZase. The purpose of this study was to understand the molecular basis and the characteristics of pncA gene mutations and its relation to PZA resistance in M. tuberculosis strains from China. Several mutations with base changes leading to amino acid substitutions were found in the PZA-resistant isolates. No mutations were seen in the 243 PZA-susceptible isolates. Among the 35 PZA-resistant isolates, 32 isolates (91.4%) had nucleotide substitutions, insertions and deletions that resulted in amino-acid substitution; or frameshifts in some strains. Other previously uncharacterized mutations were found as follows: Asn118->Thr, CG insertion at position 501; CC insertion at nucleotide position 403; a 8 base-pair deletion at start codon; Pro54->Thr; AG insertion at 368; Tyr41->His, Ser88->stop, and A insertion at nucleotide position 301. IS6110 subtyping revealed that each strain was unique; indicative of the epidemiologic independence of the isolates. PMID- 10813148 TI - The stethoscope in the Emergency Department: a vector of infection? AB - The purposes of this study were to determine whether microorganisms can be isolated from the membranes of stethoscopes used by clinicians and nurses, and to analyse whether or not the degree of bacterial colonization could be reduced with different cleaning methods. We designed a transversal before-after study in which 122 stethoscopes were examined. Coagulase negative staphylococci (which are also potentially pathogenic microorganisms) were isolated together with 13 other potentially pathogenic microorganisms, including S. aureus, Acinetobacter sp. and Enterobacter agglomerans. The most effective antiseptic was propyl alcohol. Analysis of the cleaning habits of the Emergency Department (ED) staff, showed that 45% cleaned the stethoscope annually or never. The isolation of potentially pathogenic microorganisms suggests that the stethoscope must be considered as a potential vector of infection not only in the ED but also in other hospital wards and out-patient clinics. PMID- 10813149 TI - Cumulative incidence of rheumatic fever in an endemic region: a guide to the susceptibility of the population? AB - Aboriginal Australians in northern Australia are subject to endemic infection with group A streptococci, with correspondingly high rates of acute rheumatic fever and rheumatic heart disease. For 12 communities with good ascertainment, the estimated lifetime cumulative incidence of acute rheumatic fever was approximately 5.7%, whereas over the whole population, with less adequate ascertainment, the cumulative incidence was only 2.7%. The corresponding prevalences of established rheumatic heart disease were substantially less than the cumulative incidences of acute rheumatic fever, at least in part because of poor ascertainment. The cumulative incidence of acute rheumatic fever estimates the proportion of susceptible individuals in endemically exposed populations. Our figures of 2.7-5.7% susceptible are consistent with others in the literature. Such comparisons suggest that the major part of the variation in rheumatic fever incidence between populations is due to differences in streptococcal exposure and treatment, rather than to any difference in (genetic) susceptibility. PMID- 10813150 TI - Population estimates of persons presenting to general practitioners with influenza-like illness, 1987-96: a study of the demography of influenza-like illness in sentinel practice networks in England and Wales, and in The Netherlands. AB - Incidence data by age of new episodes of influenza-like illness reported by sentinel general practice networks in England and Wales and in The Netherlands over a 10-year period were examined to provide estimates of the consulting population during influenza epidemic periods. Baseline levels of recording in each age group were calculated from weeks in which influenza viruses were not circulating and the excess over baseline calculated to provide the population estimates during influenza epidemics. Influenza A/H3N2 epidemics were associated with higher population estimates for consultations than influenza B, especially in the age groups 0-4 and 65 years and over. In the intervening age groups, population estimates were more consistent regardless of the virus type. Both networks reported simultaneous peaking of incidence rates in all of the age groups. There were substantial increases in the number of persons reporting other respiratory illnesses during influenza epidemics. Population estimates of the consulting population provide the only secure basis for which health services resource utilization during influenza epidemics can be estimated. PMID- 10813151 TI - Non-participation in a population-based seroprevalence study of vaccine preventable diseases. AB - To estimate the immunity of the Dutch population against vaccine-preventable diseases, a population-based serum bank was established. Since a multi-tiered approach to enroll eligible individuals was used, both the overall non-response selection and the effect, on this selection, of including additional participants and of excluding a subgroup of non-participants (i.e. those without questionnaire data) could be studied. For some characteristics associated with non participation, an association with seroprevalence of vaccine-preventable diseases is likely (e.g. age, gender). For other characteristics (e.g. marital status, receipt of reminder, degree of urbanization) the association with immune status is unclear but probably small. If the distribution in the population, or information on all participants and non-participants, of the characteristic is available, then the effect on the seroprevalence can be estimated. However, investigators have to be aware that studying only a subgroup of non-participants might lead to a biased insight into non-participation selection. Furthermore, merely including additional participants might not always reduce this bias. PMID- 10813152 TI - Secondary measles vaccine failures identified by measurement of IgG avidity: high occurrence among teenagers vaccinated at a young age. AB - Failure to seroconvert (primary vaccine failure) is believed to be the principal reason (approx. > 95%) why some vaccinees remain susceptible to measles and is often attributed to the persistence of maternal antibodies in children vaccinated at a young age. Avidity testing is able to separate primary from secondary vaccine failures (waning and/or incomplete immunity), but has not been utilized in measles epidemiology. Low-avidity (LA) and high-avidity (HA) virus-specific IgG antibodies indicate primary and secondary failure, respectively. Measles vaccine failures (n = 142; mean age 10.1 years, range 2-22 years) from an outbreak in 1988-9 in Finland were tested for measles-virus IgG avidity using a protein denaturating EIA. Severity of measles was recorded in 89 failures and 169 non-vaccinees (mean age 16.2 years, range 2-22 years). The patients with HA antibodies (n = 28) tended to have clinically mild measles and rapid IgG response. Among failures vaccinated at < 12, 12-15 and > 15 months of age with single doses of Schwarz-strain vaccine in the 1970s, 50 (95% CI 1-99), 36 (CI 16 56) and 25% (CI 8-42) had HA antibodies, respectively. When a single measles, mumps and rubella (MMR) vaccine had been given after 1982 at 15 months of age, only 7% (CI 0-14) showed HA antibodies. Omitting re-vaccinees and those vaccinated at < 15 months, Schwarz-strain recipients had 3.6 (CI 1.1-11.5) higher occurrence of HA responses compared to MMR recipients. Apart from one municipality, where even re-vaccinees had high risk of primary infection, 89% (CI 69 to approximately 100) of the infected re-vaccinees had an HA response. Secondary measles-vaccine failures are more common than was more previously thought, particularly among individuals vaccinated in early life, long ago, and among re-vaccinees. Waning immunity even among individuals vaccinated after 15 months of age, without the boosting effect of natural infections should be considered a relevant possibility in future planning of vaccination against measles. PMID- 10813153 TI - Estimation of the basic reproduction number of measles during an outbreak in a partially vaccinated population. AB - From March to July 1996 a measles outbreak occurred in northern Luxembourg with 110 reported cases centered around two primary schools (85 cases) and the surrounding community (25 cases). Eighty four suspected cases were confirmed serologically. Vaccine coverage was estimated from questionnaire-based surveys at the two primary schools to be 70 and 76%, respectively. Vaccine efficacy during the outbreak was estimated to be 94.6% [95% confidence interval (CI) 90.4-97.0]. Using the information from the, school surveys, we obtained estimates of the basic reproduction number of measles of 7.7 (95% CI 4.4-11.0) and 6.2 (95% CI 3.5 8.9), respectively. Assuming a 95% vaccine efficacy, these estimates correspond to minimal vaccine coverages of 91.6% (95% CI 81.4-95.7) and 88.3% (95% CI 75.5 93.4) which would have been necessary to minimize the chances of a major outbreak occurring. We can confirm that major outbreaks in similar school settings can only be prevented if vaccination coverage exceeds 90%. PMID- 10813154 TI - Predicting and preventing measles epidemics in New Zealand: application of a mathematical model. AB - A mathematical model of the dynamics of measles in New Zealand was developed in 1996. The model successfully predicted an epidemic in 1997 and was instrumental in the decision to carry out an intensive MMR (measles-mumps rubella) immunization campaign in that year. While the epidemic began some months earlier than anticipated, it was rapidly brought under control, and its impact on the population was much reduced. In order to prevent the occurrence of further epidemics in New Zealand, an extended version of the model has since been developed and applied to the critical question of the optimal timing of MMR immunization. PMID- 10813155 TI - Measles resurgence in Argentina: 1997-8 outbreak. AB - Epidemiological and clinical findings from 1162 serologically confirmed measles cases occurring in Buenos Aires, Argentina in 1997 and 1998 were retrospectively reviewed. From 90 hospitalized children, measles virus was detected by direct RT PCR from nasopharyngeal secretions. Patients were grouped as follows: (i) not vaccinated: infants < 12 months; (ii) regularly vaccinated: children 1-4 years not covered by the last catch-up; (iii) catch-up vaccinated: patients 5-19 years immunized during the 1993 campaign. Most cases were recorded in non-vaccinated infants (54%), and the lowest in catch-up vaccinated children (16%). Mean age of the 90 hospitalized children was 11.3 months. Pneumonia was the major hospitalization cause followed by pneumonitis. Two children required intensive care and one died. The 1993 catch-up campaign seemed to reduce the number of cases in the 5- to 19-year-old group. Lack of timely follow-up probably led to the accumulation of susceptible individuals allowing measles re-emergence. Direct viral detection by RT-PCR proved to be a sensitive tool for molecular epidemiology surveillance. PMID- 10813156 TI - Predictions of the emergence of vaccine-resistant hepatitis B in The Gambia using a mathematical model. AB - Vaccine escape variants of hepatitis B virus (HBV) have been identified world wide. A mathematical model of HBV transmission is used to investigate the potential pattern of emergence of such variants. Attention is focused on The Gambia as a country with high quality epidemiological data, universal infant immunization and in which escape mutants after childhood infections have been observed. We predict that a variant cannot become dominant for at least 20 years from the start of vaccination, even when using a vaccine which affords no cross protection. The dominant factor responsible for this long time scale is the low rate of infectious contacts between infected and susceptible individuals (we estimate the basic reproduction number of hepatitis B in The Gambia to be 1.7). A variant strain that achieves high prevalence will also take many years to control, and it is questionable whether emergence will be identifiable by sero surveillance until of high prevalence. The sensitivity of the model predictions to epidemiological and demographic factors is explored. PMID- 10813157 TI - A community outbreak of hepatitis A in a religious community in Indiana: failure of immune serum globulin to prevent the spread of infection. AB - An outbreak of hepatitis A occurred in a religious community in Indiana, USA. Sixty-nine cases were ascertained among the 4466 residents over a year, and the highest attack rate was in children. The management of the outbreak included the widespread use of prophylactic immune serum globulin (ISG). Despite this, further cases occurred. To guide further ISG administration, a survey was undertaken to ascertain what proportion remained susceptible to HAV infection. From a random sample of 600 people in the affected community 440 saliva specimens (73%) were obtained. Of these, 12.5% were found to be immune (95% confidence intervals from 9-16%). No changes were made to the ISG administration policy. There was no evidence to suggest that administration of ISG had any effect on the duration of the outbreak. There was a low rate of symptomatic infection among young children (less than 10 years); as ISG does not prevent the spread of the virus its use is not recommended in future outbreak situations. PMID- 10813158 TI - Field evidence that roe deer (Capreolus capreolus) are a natural host for Ehrlichia phagocytophila. AB - Samples of blood, spleen and legs from 112 culled roe deer (Capreolus capreolus) were collected from nine sites widespread in the United Kingdom. The prevalence of infection with Ehrlichia phagocytophila was determined by serology and polymerase chain reaction. Means of 58% of 102 plasma or serum samples were seroreactive by IFA, 38% of 84 blood samples and 29% of 82 spleen samples were positive by PCR. Ticks on legs of 71 roe deer were Ixodes ricinus larvae, nymphs and adults and 83% of legs were infested. Numbers of ticks corresponded positively to the percentage of samples positive for E. phagocytophila by serology and PCR for different sampling sites. Ixodes ricinus nymphs collected from the vegetation at one site with infected deer were analysed for infection with E. phagocytophila by examination of Feulgen stained salivary glands. Of 135 nymphs 5% were infected. These results confirm that roe deer are commonly parasitized by both E. phagocytophila and its vector tick in such a way that it is likely to be an important natural mammalian reservoir of E. phagocytophila. PMID- 10813159 TI - Differences in the epidemiology of theileriosis on smallholder dairy farms in contrasting agro-ecological and grazing strata of highland Kenya. AB - A prospective cohort study was conducted in five purposively-sampled agro ecological zone (AEZ)-grazing system strata in Murang'a District, Kenya, between March 1995 and June 1996. The study strata were selected based on a preliminary characterization study to represent the widest range of risks to East Coast fever (ECF) in the District and included zero-grazing and open-grazing farms. In total, 225 calves from 188 smallholder farms were examined from birth to 6 months of age and visited within the first 2 weeks of life and thereafter at bi-weekly intervals for up to 14 visits. The purpose of the study was to characterize the differences in epidemiology (risks of infection, morbidity and mortality) and potential control of ECF between the selected strata. Evidence of Theileria parva infection was assessed by increased antibody levels as measured in an indirect ELISA assay by the percent positivity (PP) of serum samples relative to a strong positive reference serum. Sero-conversion risks of T. parva were highest in the open-grazing strata. Antibody prevalence in adult cattle and ECF morbidity and mortality risks were also highest in open-grazing strata. While different, all five AEZ-grazing strata were considered to be endemically unstable for ECF. East Coast fever challenge was low in all zero-grazing strata and this challenge is likely to remain low due to continuing intensification of smallholder farming in the central highlands. In the open-grazing strata, there was higher challenge and a greater impact of ECF. PMID- 10813160 TI - Distribution of the meningococcal insertion sequence IS1301 in clonal lineages of Neisseria meningitidis. AB - The distribution of the meningococcal insertion sequence IS1301 was analysed in 496 strains of different serogroups and clonal lineages of Neisseria meningitidis, and in 64 neisserial strains other than N. meningitidis. IS1301 was found in meningococci, but not in apathogenic Neisseria sp. and Neisseria gonorrhoeae. The copy numbers of IS1301 varied between 2 and 17 per genome. IS1301 positive strains were mostly found among the serogroups 29E, W135, X, and Y. Clonal lineages of serogroup A, B, and C meningococci associated with epidemic meningococcal disease were rarely positive for IS1301. PMID- 10813161 TI - Risk of transmission of blood-borne viral infection in orthopaedic and trauma surgery. PMID- 10813162 TI - Positron emission tomography and the orthopaedic surgeon. PMID- 10813163 TI - Analysis of muscle function in the lower limb after fracture of the diaphysis of the tibia in adults. AB - We examined the recovery of power in the muscles of the lower limb after fracture of the tibial diaphysis, using a Biodex dynamometer. Recovery in all muscle groups was rapid for 15 to 20 weeks following fracture after which it slowed. Two weeks after fracture the knee flexors and extensors have about 40% of normal power, which rises to 75% to 85% after one year. The dorsiflexors and plantar flexors of the ankle and the invertors and evertors of the subtalar joint are much weaker two weeks after injury, but at one year their mean power is more than that of the knee flexors and extensors. Our findings showed that age, the mode of injury, fracture morphology, the presence of an open wound and the Tscherne grade of closed fractures correlated with muscle power. It is age, however, which mainly determines muscle recovery after fracture of the tibial diaphysis. PMID- 10813164 TI - Treatment of nonunion around the olecranon fossa of the humerus by intramedullary locked nailing. AB - Nonunion of fractures of the olecranon fossa of the humerus presents a difficult surgical problem. The distal fragment is usually small and osteoporotic and stable fixation is not easy to achieve. We describe a modification of the technique of locked nailing by which the distal aspect of the nail is placed in the subchondral region of the trochlea. Good results were obtained in seven out of eight patients with this technique. PMID- 10813165 TI - Fixation of fractures of the shaft of the humerus by dynamic compression plate or intramedullary nail. A prospective, randomised trial. AB - We randomised prospectively 44 patients with fractures of the shaft of the humerus to open reduction and internal fixation by either an intramedullary nail (IMN) or a dynamic compression plate (DCP). Patients were followed up for a minimum of six months. There were no significant differences in the function of the shoulder and elbow, as determined by the American Shoulder and Elbow Surgeons' score, the visual analogue pain score, range of movement, or the time taken to return to normal activity. There was a single case of shoulder impingement in the DCP group and six in the IMN group. Of these six, five occurred after antegrade insertion of an IMN. In the DCP group three patients developed complications, compared with 13 in the IMN group. We had to perform secondary surgery on seven patients in the IMN group, but on only one in the DCP group (p = 0.016). Our findings suggest that open reduction and internal fixation with a DCP remains the best treatment for unstable fractures of the shaft of the humerus. Fixation by IMN may be indicated for specific situations, but is technically more demanding and has a higher rate of complications. PMID- 10813166 TI - Fractures of the distal radius treated by internal fixation and early function. A prospective study of 73 consecutive patients. AB - Stable fixation of fractures of the distal radius can be achieved by using two 2.0 mm titanium plates placed on the radial and intermediate columns angled 50 degrees to 70 degrees apart. We describe our results with this method in a prospective series of 74 fractures (58 severely comminuted) in 73 consecutive patients. Early postoperative mobilisation was possible in all except four wrists. All of the 73 patients, except two with other injuries, returned to work and daily activities with no limitations. The anatomical results were excellent or good in 72 patients and fair in one. Our discussion includes details of important technical considerations based on an analysis of the specific complications which were seen early in the series. PMID- 10813167 TI - Arthroscopic findings in acute fractures of the ankle. AB - We have evaluated prospectively the arthroscopic findings in acute fractures of the ankle in 288 consecutive patients (148 men and 140 women) with a mean age of 45.6 years. According to the AO-Danis-Weber classification there were 14 type-A fractures, 198 type B and 76 type C. Lesions of the cartilage were found in 228 ankles (79.2%), more often on the talus (69.4%) than on the distal tibia (45.8%), the fibula (45.1%), or the medial malleolus (41.3%). There were more lesions in men than in women and in general they were more severe in men (p < 0.05). They also tended to be worse in patients under 30 years and in those over 60 years of age. The frequency and severity of the lesions increased from type-B to type-C fractures (p < 0.05). Within each type of fracture the lesions increased from subgroups 1 to 3 (p < 0.05). The anterior tibiofibular ligament was injured with increased frequency from type-B.1 to type-C3 fractures (p < 0.05), but it was not torn in all cases. While lateral ligamentous injuries were seen more often in type-B than in type-C fractures (p < 0.05), no difference was noted in the frequency of deltoid ligamentous lesions. Our findings show that arthroscopy is useful in identifying associated intra-articular lesions in acute fractures of the ankle. PMID- 10813169 TI - The anatomy and function of the gluteus minimus muscle. AB - In order to investigate the functional anatomy of gluteus minimus we dissected 16 hips in fresh cadavers. The muscle originates from the external aspect of the ilium, between the anterior and inferior gluteal lines, and also at the sciatic notch from the inside of the pelvis where it protects the superior gluteal nerve and artery. It inserts anterosuperiorly into the capsule of the hip and continues to its main insertion on the greater trochanter. Based on these anatomical findings, a model was developed using plastic bones. A study of its mechanics showed that gluteus minimus acts as a flexor, an abductor and an internal or external rotator, depending on the position of the femur and which part of the muscle is active. It follows that one of its functions is to stabilise the head of the femur in the acetabulum by tightening the capsule and applying pressure on the head. Careful preservation or reattachment of the tendon of gluteus minimus during surgery on the hip is strongly recommended. PMID- 10813168 TI - Extensive metallosis and necrosis in failed prostheses with cemented titanium alloy stems and ceramic heads. AB - We describe three prostheses with cemented titanium-alloy stems and Al2O3 ceramic femoral heads which had to be revised after a mean period of implantation of 78 months. In each case, the neck of the prosthesis had been so severely worn that the profile was elliptical rather than circular. There was severe metallosis of the periprosthetic tissues. Metal particles isolated from the tissues were approximately one nanometre in size and the ratios of titanium, aluminium and vanadium in the particles were the same as in the original alloy. Histologically, the high concentration of metal particles masked the presence of high-density polyethylene (HDP) debris, but again particles about one nanometre in size were isolated from the tissues. The severe necrobiosis and necrosis noted were consistent with other reports of the presence of extensive wear particles in periprosthetic tissues. Wear is presumed to have occurred as a result of mismatch between the shape or size of the taper cone and the femoral head, or to changes in the geometry of loading due to migration of the cup. To facilitate early intervention, patients with this design of prosthesis should be monitored radiologically. PMID- 10813170 TI - Radial MRI of the hip with moderate osteoarthritis. AB - We carried out radial MRI in 30 hips with moderate osteoarthritis and in ten normal hips. On a scout view containing the entire acetabular rim, 12 vertical radial slices were set at 15 degrees intervals. Different appearances were observed in different parts of the joint. In the weight-bearing portion, from 45 degrees anterosuperior to 45 degrees posterosuperior, 'attenuation' (n = 16) and 'disappearance' (n = 25) were observed as abnormalities of the labrum with 'capsular stripping' (n = 29) and 'extraosseous high signal lesion' (n = 27) as capsular abnormalities, seen more often in the anterosuperior portion. In all 12 planes there were osteophytes on the acetabular edge (n = 24), femoral head (n = 22) and/or at the central acetabulum (n = 6), a bone cyst on the acetabulum (n = 18) and/or the femoral head (n = 9), irregularity of the articular cartilage (n = 30), and an effusion (n = 28). Our findings indicate that radial MRI may be a useful non-invasive diagnostic method for demonstrating pathology in moderate osteoarthritis of the hip. PMID- 10813171 TI - Factors influencing joint-preserving operations in the treatment of the late stages of osteoarthritis of the hip. AB - Between November 1983 and December 1992, 136 hips (119 patients) with coxarthritis were operated on using joint-preserving techniques based on the rationale of Pauwels' osteotomy. The criterion for selection was a patient in whom the height of the joint space in the weight-bearing area of the hip was less than 1 mm. The mean age at operation was 48 years and the mean follow-up 109 months (60 to 171). Hips were categorised using Bombelli's classification of osteoarthritis, into atrophic and non-atrophic types. The endpoint was defined as that at which the height of the joint space became less than 1 mm again. The Kaplan-Meier curve showed that the rate of survival of the non-atrophic group was significantly better than that of the atrophic group. Cox's proportional hazard model indicated that the factors influencing the results of joint-preserving operations included Bombelli's classification, postoperative incongruence of the joint and the height of the joint space. PMID- 10813172 TI - Cemented total hip arthroplasty with autogenous acetabular bone grafting for hips with developmental dysplasia in adults: the results at a minimum of ten years. AB - We carried out primary cemented total hip arthroplasty (THA) on 25 hips in 21 patients with developmental dysplasia of the hip, using autogenous acetabular bone grafts. The socket was placed at the level of the true acetabulum and bone from the femoral neck was used as graft. Five hips were excluded, leaving 20 which were followed up for a mean of 12.9 years (10 to 18). The mean modified Merle d'Aubigne and Postel functional scores were 5.6 for pain, 4.3 for mobility and 4.2 for range of movement. Radiological examination showed aseptic loosening in three sockets but not in the stems. The bone grafts had united and showed no evidence of late failure. PMID- 10813173 TI - The results at nine to twelve years of the use of a hydroxyapatite-coated femoral stem. AB - We describe the clinical and radiological outcome of 100 consecutive total hip replacements in 86 patients using the JRI Furlong hydroxyapatite-coated femoral component. The follow-up was 100% at a mean of ten years (9.0 to 12.0). All 54 living patients (62 hips) were examined annually. Their mean Merle d'Aubigne score was 5.8/5.6/4.6. A total of 32 patients had died. Their notes at the last annual assessment recorded a similar outcome to that of the living patients. Radiographs showed no radiolucent lines around any femoral component and there were no signs of impending failure. One patient required excision arthroplasty, but the femoral component had been bonded satisfactorily. There have been no other revisions of femoral components. Our findings suggest that the hydroxyapatite coating gives a satisfactory prosthesis-bone interface which is preferable to any other system. PMID- 10813174 TI - The orthopaedic management of peripheral ischaemia in meningococcal septicaemia in children. AB - Between March 1993 and February 1999, 14 children aged from eight months to 14.75 years were admitted to the paediatric intensive-care unit with meningococcal septicaemia in association with severe peripheral ischaemia. Of these, 13 were operated upon, eight of whom had early fasciotomies. Five children died. Of the nine survivors, one had no amputations while in the other eight 14 limb segments were amputated. We review the case histories and propose a protocol for the early management of these children. PMID- 10813175 TI - The Ilizarov method in the management of relapsed club feet. AB - We present the results of the management of 17 relapsed club feet in 12 children using the Ilizarov method with gradual distraction and realignment of the joint. Review at a mean of three years after surgery showed maintenance of correction with excellent or good results in 13 feet. Five mobile feet which had been treated by a split transfer of the tibialis anterior tendon two weeks after removal of the frame had an excellent result. PMID- 10813176 TI - The blood supply of the lateral epiphyseal arteries in Perthes' disease. AB - We performed superselective angiography in 28 hips in 25 patients with Perthes' disease in order to study the blood supply of the lateral epiphyseal arteries (LEAs). Interruption of the LEAs at their origin was observed in 19 hips (68%). Revascularisation in the form of numerous small arteries was seen in ten out of 11 hips in the initial stage of Perthes' disease, in seven of eight in the fragmentation stage and in five of nine in the healing stage. Penetration of mature arteries into the depths of the epiphysis was seen in four of nine hips in the healing stage. Vascular penetration was absent in the weight-bearing portion of the femoral head below the acetabular roof. Interruption of the posterior column artery was seen where it passed through the capsule in seven hips when they lay either in internal rotation or in abduction with internal rotation. We suggest that in Perthes' disease the blood supply of the LEAs is impaired at their origin and that revascularisation occurs from this site by ingrowth of small vessels into the femoral epiphysis. This process may be the result of recurrent ischaemic episodes. PMID- 10813177 TI - Melatonin and adolescent idiopathic scoliosis. AB - Scoliosis seen in the chicken after pinealectomy resembles adolescent idiopathic scoliosis in man. It has been suggested that in both species, deficiency of the pineal hormone, melatonin, is responsible for this phenomenon. In nine patients with adolescent idiopathic scoliosis and in ten age- and gender-matched controls, the circadian levels of serum melatonin and the excretion of urinary 6-hydroxy melatonin-sulphate, the principal metabolite of melatonin, were determined. There were no statistically significant differences in the secretion of serum melatonin or the excretion of urinary 6-hydroxy-melatonin-sulphate between the patients and the control group. The hypothesis of melatonin deficiency as a causative factor in the aetiology of adolescent idiopathic scoliosis cannot be supported by our data. PMID- 10813178 TI - The 'chef's hat' appearance of the femoral head in cleidocranial dysplasia. AB - Cleidocranial dysplasia (CCD) is inherited as an autosomal dominant disorder characterised by failure of membranous ossification. The condition is due to a mutation of the cbfa1 gene on chromosome 6 which has a role in the development of osteoblasts from the mesenchymal cells. In their growing years, these patients have an unusual shape of the femoral head reminiscent of a 'chef's hat'. In order to confirm the consistency of this sign, we have reviewed the radiographs of 28 patients with CCD. All except three had this appearance. The sign was also seen in patients with coxa vara associated with a variety of other conditions. The chef's hat sign may occur secondary to the particular mechanical environment created by coxa vara as well as abnormal cellular function in patients with CCD. Although coxa vara has some influence on the shape of the femoral head, it is not entirely responsible for its morphology since it was present in only six of the 28 patients with CCD. PMID- 10813179 TI - Vascularised bone grafting for fibrous dysplasia of the upper limb. AB - We describe our experience with vascularised bone grafting for the treatment of fibrous dysplasia of the upper limb in eight patients, five men and three women, aged between 17 and 36 years. The site was in the humerus in six and the radius in two. Persistent pain, progression of the lesion and pathological fracture with delayed union were the indications for surgical intervention. We used a vascularised fibular graft after curettage of the lesion. Function and radiological progress were serially monitored. Early radiological union of the graft occurred at periods ranging from 8 to 14 weeks. The mean period for reconstitution of the diameter of the bone was 14 months (12 to 18) predominantly through inductive formation of bone around the vascularised graft, which was a prominent feature in all patients. There were no recurrences and none of the grafts sustained a fracture or failed to unite. After operation function was excellent in three patients and good in five. Vascularised bone grafts provide a safe and reliable means of ensuring good continuity of bone with little risk of recurrence and failure. PMID- 10813180 TI - Cryosurgery for chronic injuries of the cutaneous nerve in the upper limb. Analysis of a new open technique. AB - We have treated six patients with chronic pain following nerve injury using a cryosurgical probe. All had a significant return of hand function and improvement of pain during a mean follow-up of 13.5 months. Open visualisation of the injured nervous tissue is essential for patients undergoing this technique. Four patients regained normal sensation in the dermatome of the previously injured nerve. PMID- 10813181 TI - The depressor function of biceps on the head of the humerus in shoulders with tears of the rotator cuff. AB - We investigated the function of biceps in 18 patients (19 shoulders) with lesions of the rotator cuff. Their mean age was 59 years. Another series of 18 patients (19 shoulders) with normal rotator cuffs as seen on MRI acted as a control group. Their mean age was 55 years. A brace was used to maintain contraction of biceps during elevation. Anteroposterior radiographs were obtained with the arm elevated at 0 degrees , 45 degrees and 90 degrees with and without contraction of biceps. The distance between the centre of the head of the humerus and the glenoid was compared in the two groups. We found that in the group with tears there was significantly greater proximal migration of the head of the humerus at 0 degrees and 45 degrees of elevation without contraction of biceps but depression of the head of the humerus at 0 degrees, 45 degrees and 90 degrees when biceps was functioning. We conclude that biceps is an active depressor of the head of the humerus in shoulders with lesions of the rotator cuff. PMID- 10813182 TI - Surgical treatment of compression of the lateral antebrachial cutaneous nerve. AB - We describe an operation to relieve compression of the lateral antebrachial cutaneous nerve at the elbow. Between 1987 and 1997 we operated on seven patients, one with bilateral compression. In two the compression was associated with injury to biceps. A longitudinal or a transverse incision was carried out and the nerve was released from the deep fascia. Partial excision of the biceps aponeurosis was undertaken in the patients who did not have injury to biceps; some additional procedures were required for those patients with injuries. All patients had symptomatic relief. PMID- 10813183 TI - A retinacular sling for subluxing tendons of the first extensor compartment. A case report. AB - Over-zealous release of the first dorsal compartment of the wrist for de Quervain's disease or other lesions such as ganglia, may result in volar subluxation of the tendons of abductor pollicis longus and extensor pollicis brevis. This is usually asymptomatic, but may occasionally become disabling. We describe an operation using part of the extensor retinaculum to stabilise such a subluxation. PMID- 10813184 TI - Magnetic resonance arthrography of the acetabular labrum. Macroscopic and histological correlation in 20 cadavers. AB - We studied the sensitivity and specificity of magnetic resonance arthrography (MRa) for the diagnosis of lesions of the acetabular labrum in 20 cadaver hips. The MRa results were compared with macroscopic and histological findings. We found that the labrum could be satisfactorily delineated by MRa and that large detachments could be identified satisfactorily. The diagnosis of small detachments and degeneration of the labrum was less reliable. PMID- 10813185 TI - The effect of the position of the limb on venous impulse foot pumps. AB - Compression foot pumps are widely used for the prevention of postoperative venous thrombosis. We tested the efficiency of the pump in ten healthy subjects; the velocity of venous blood flow in the common femoral vein was measured in the horizontal, Trendelenberg (foot-up) and reverse-Trendelenberg (foot-down) positions. Application of the foot pump produced an increase in the venous velocity in all subjects. The mean increase in the horizontal position was 27.2% and in the Trendelenberg position 15.4%. In the reverse-Trendelenberg position, the foot pump produced a mean increase of 102.8%. The efficiency of the compression foot pump in increasing venous return is improved by adopting the reverse-Trendelenberg position. This may increase its thromboprophylactic effect. PMID- 10813186 TI - Autologous morsellised bone grafting restores uncontained femoral bone defects in knee arthroplasty. An in vivo study in horses. AB - The properties of impacted morsellised bone graft (MBG) in revision total knee arthroplasty (TKA) were studied in 12 horses. The left hind metatarsophalangeal joint was replaced by a human TKA. The horses were then randomly divided into graft and control groups. In the graft group, a unicondylar, lateral uncontained defect was created in the third metatarsal bone and reconstructed using autologous MBG before cementing the TKA. In the control group, a cemented TKA was implanted without the bone resection and grafting procedure. After four to eight months, the animals were killed and a biomechanical loading test was performed with a cyclic load equivalent to the horse's body-weight to study mechanical stability. After removal of the prosthesis, the distal third metatarsal bone was studied radiologically, histologically and by quantitative and micro CT. Biomechanical testing showed that the differences in deformation between the graft and the control condyles were not significant for either elastic or time dependent deformations. The differences in bone mineral density (BMD) between the graft and the control condyles were not significant. The BMD of the MBG was significantly lower than that in the other regions in the same limb. Micro CT showed a significant difference in the degree of anisotropy between the graft and host bone, even although the structure of the area of the MBG had trabecular orientation in the direction of the axial load. Histological analysis revealed that all the grafts were revascularised and completely incorporated into a new trabecular structure with few or no remnants of graft. Our study provides a basis for the clinical application of this technique with MBG in revision TKA. PMID- 10813187 TI - Changes in the loads on an internal spinal fixator after iliac-crest autograft. AB - Spines are often stabilised posteriorly by internal fixation and anteriorly by a bone graft. The effect of an autologous bone graft from the iliac crest on implant loads is unknown. We used an internal spinal fixation device with telemetry to measure implant loads for several body positions and activities in nine patients before and after anterior interbody fusion. With the body upright, implant loads were often higher after than before fusion using a bone graft. Distraction of the bridged region led to high implant loads in patients with a fractured vertebra and to marked changes in load in those with degenerative instability. Leaving the lower of the bridged intervertebral discs intact led to only small changes in fixator load after anterior interbody fusion. A bone graft alone does not guarantee a reduction of implant loads. PMID- 10813188 TI - Macro and microscopic examination of the ruptured surfaces of anterior cruciate ligaments of rabbits. AB - In a study combining tissue mechanics and fracture morphology for the first time, we examined the ruptured surfaces of anterior cruciate ligaments of rabbits and related their appearance to the initial loading conditions. Sixteen specimens were stretched to failure at rates of displacement of 10 and 500 mm/min. We used video images to study the changes which occurred during the fracture process and SEM to examine the appearance of the ruptured surfaces. The surfaces of ligaments tested at 10 mm/min had more pulled-out collagen fibres and the fibres had more pronounced waviness compared with those tested at 500 mm/min. We have shown that the macroscopic appearance of ruptured ligaments can be related to their microscopic appearance and that it is possible to deduce whether failure was by gradual tearing of the fibres or catastrophic failure. PMID- 10813189 TI - Hydroxyapatite coated with heaptocyte growth factor (HGF) stimulates human osteoblasts in vitro. AB - We have studied in vitro the effect of a hydroxyapatite (HA) tricalcium phosphate material coated with hepatocyte growth factor (HA-HGF) on cell growth, collagen synthesis and secretion of metalloproteinases (MMPs) by human osteoblasts. Cell proliferation was stimulated when osteoblasts were incubated with untreated HA and was further increased after exposure to HA-HGF. The uptake of [3H]-proline was increased after treatment with HA. When osteoblasts were exposed to HA-HGF, collagen synthesis was increased with respect to HA. The secretion of MMPs in control cells was undetectable, but in HA and HA-HGF cells MMP 2 and MMP 9 were clearly synthesised. Our results suggest that HA can promote osteoblast activity and that HGF can further increase its bioactivity. PMID- 10813190 TI - Percutaneous repair of the ruptured tendo Achillis. PMID- 10813191 TI - Work practice and histopathological changes in the tenosynovium and flexor retinaculum in carpal tunnel syndrome in women. PMID- 10813192 TI - Disarticulation at the ankle using an anterior flap. PMID- 10813193 TI - Plain radiography in the degenerate knee. PMID- 10813194 TI - Biomechanical comparison of fixation of type-I fractures of the lateral tibial plateau. PMID- 10813195 TI - Screw versus suture fixation of Mitchell's osteotomy. PMID- 10813196 TI - Nonunion of tibial stress fractures in patients with deformed arthritic knees. PMID- 10813197 TI - The problem of diffuse alveolar hemorrhage syndromes. AB - No matter how little the amount of blood and no matter how infrequent the event, the presence of hemoptysis is a terrifying and infernal experience for the patient. For the physician the evaluation of pulmonary bleeding carries an urgency that requires a logical plan and prompt execution. PMID- 10813198 TI - Nitric oxide and interstitial lung disease. AB - The role and significance of nitric oxide (NO) and its intermediates on interstitial lung diseases are discussed. NO itself has protective roles against lung injury; however, NO is converted to peroxynitrite through the reaction with the superoxide anion at the inflammation foci, resulting in various destructive effects against lung parenchyma. Increased production of NO and peroxynitrite play an important role in oxidative injury and remodeling associated with interstitial lung diseases. PMID- 10813199 TI - Classification and recent advances in idiopathic interstitial pneumonia. AB - Idiopathic interstitial pneumonia (IIP) is a heterogeneous group of diseases comprising acute interstitial pneumonia, bronchiolitis obliterans organizing pneumonia (BOOP), nonspecific interstitial pneumonia, desquamative interstitial pneumonia, and idiopathic pulmonary fibrosis and usual interstitial pneumonia (IPF/UIP). We review the clinicopathological spectrum of IIP and introduce recent advances in classification, treatment, and prognosis. BOOP can be clinically categorized as an interstitial pneumonia, though prominent granulation tufts are seen in the airspaces. Though differences between the nonspecific interstitial pneumonia and other lips can be histopathologically clarified, the focus of clinical research on NSIP is differentiation from BOOP, or from IPF and UIP. IIP can be categorized into two groups: groups with acute or subacute lung injuries or fibrosis, such as in acute interstitial pneumonia, BOOP and nonspecific interstitial pneumonia, and groups with chronic injuries or fibrosis, such as IPF/UIP. This classification accords well with the maturity of fibrosis, CT findings, bronchoalveolar lavage fluid cell findings, and prognosis. The most critical problem is the treatment of IPF/UIP, because of its high mortality. PMID- 10813200 TI - Cell apoptosis and granulomatous lung diseases. AB - Apoptosis, also known as activation-induced cell death or programmed cell death, is an active suicide mechanism that is involved in normal tissue turnover during embryogenesis and adult life. There are many examples of apoptosis in the immune system, including programmed cell death of T cells during negative intrathymic selection of the TCR repertoire and, in the postthymic phase, death of responsive T cells upon specific activation of the TCR/CD3 complex. Induction of apoptosis assures rapid disappearance of the immune response upon antigenic clearance, avoiding the metabolic costs involved in sustaining a large number of effector cells. The knowledge that failure of immune cells to die is the cause of a number of immune-mediated disorders has opened intriguing new avenues of exploration into the pathogenetic events leading to the accumulation of immunoinflammatory cells at sites of ongoing inflammation in granulomatous disorders, including granulomas initiated by infectious agents, such as Mycobacterium tuberculosis, or in sarcoidosis. In this paper we review recent results obtained in experimental animal models and patients with immune granuloma suggesting that the positive induction by ligands binding to membrane receptors or the induction or loss of intracellular suppressor signals regulates immunoregulatory mechanisms that drive the progressive development of the granulomatous structure. The great advances in understanding how mechanisms for the activation or downregulation of apoptosis have a pathogenetic role in the outcome of granulomatous disorders are also briefly considered. PMID- 10813201 TI - Preventive therapy for steroid-induced osteoporosis in interstitial lung disease. AB - We do not have to accept bone loss and high fracture rates as inevitable consequences of high-dose glucocorticoid therapy. The assessment of the risk of osteoporosis should be an integral part of the decision to commence glucocorticoid therapy. The decision to initiate a preventive strategy should be taken on the basis of the expected risk of developing osteoporosis. This risk should be evaluated on the basis of the expected duration of corticosteroid therapy, the needed dose, the drug given, and the presence of risk factors. Pro and cons of calcium, vitamin D, inhibitors of bone resorption, stimulators of bone formation, and thiazide diuretics are described in the light of the presence of an interstitial lung disease. PMID- 10813202 TI - Exercise and interstitial lung disease. AB - Interstitial lung disease (ILD) limits exercise capacity through a variety of complex and intriguing mechanisms, including ventilatory limitation, diffusion impairment, and ventilation-perfusion derangement. Resting pulmonary function testing seldom explains the symptoms nor defines the specific pathophysiology of the individual patient. Cardiopulmonary exercise testing can elucidate the relative contributions of these mechanisms and guide therapy. A fundamental problem in ILD is one of inadequate time for lung inflation during intense exercise, resulting in dynamic hypoinflation relative to the ventilatory demand. Pulmonary rehabilitation is underused in ILD. Functional CT imaging may provide insight into the relationship between structural and physiologic abnormalities of regional pulmonary function. Recent advances in nitric oxide research will perhaps further our understanding of the basic pathophysiology of ILD and provide specific treatment for the associated pulmonary vascular disease. PMID- 10813203 TI - Recent advances in radiology of the interstitial lung disease. AB - Idiopathic interstitial pneumonias are currently classified into four categories of disease: usual, desquamative, and acute interstitial pneumonia, and nonspecific interstitial pneumonia and fibrosis. Usual interstitial pneumonia appears on high-resolution CT (HRCT) as patchy subpleural areas of ground-glass opacity, irregular lines, and honeycombing. Desquamative interstitial pneumonia presents as patchy subpleural areas of ground-glass opacity in middle and lower lung zones. Acute interstitial pneumonia presents as extensive bilateral airspace consolidation and patchy or diffuse bilateral areas of ground-glass opacity. Nonspecific interstitial pneumonia and fibrosis appears as patchy or diffuse areas of ground-glass opacity with associated areas of consolidation and irregular lines. In a subset of patients with diffuse lung disease (especially in those with chronic interstitial lung disease), accurate diagnosis can be made with HRCT findings only, without surgical biopsy. However, HRCT provides a lower level of confidence in the diagnosis of acute or subacute interstitial lung disease such as infection, diffuse alveolar damage, drug reaction, or hemorrhage. Additional expiratory HRCT scans and scans with patients prone help to narrow the differential diagnosis among various diseases and help diagnose or exclude subtle disease in the posterior part of the lung, respectively. HRCT provides a reproducible method for evaluating the global extent of disease. It also discriminates between fibrotic and reversible inflammatory diseases. PMID- 10813204 TI - Constrictive (obliterative) bronchiolitis. AB - Constrictive bronchiolitis (CB), also termed in lung transplant patients obliterative bronchiolitis, is inflammation and fibrosis occurring predominantly in the walls and contiguous tissues of membranous and respiratory bronchioles with resultant narrowing of their lumens. CB is found in a variety of settings, most often as a complication of lung and heart-lung transplantation (affecting 34% to 39% of patients, usually in the first 2 years after transplantation) and bone marrow transplantation, but also in rheumatoid arthritis, after inhalation of toxic agents such as nitrogen dioxide, after ingestion of certain drugs such as penicillamine and ingestion of the East Asian vegetable Sauropus androgynous, and as a rare complication of adenovirus, influenza type A, measles, and Mycoplasma pneumoniae infections in children. In lung transplants, CB is the single most important factor leading to death thereafter. In one study, the overall mortality rate was 25%. However, at the same time, 87% of patients who were asymptomatic and diagnosed solely by transbronchial biopsy had resolution or stabilization of disease. Decreases in FEV1 from baseline can be used to clinically support CB in transplant patients; the term bronchiolitis obliterans syndrome is used to denote this clinical dysfunction, and a grading system has been established for it that is now widely used in the literature. Significant risk factors for the development of CB in lung transplants include alloantigen dependent and -independent mechanisms. In the former group are late acute rejection and HLA mismatches at the A loci; in the latter are ischemia/reperfusion injuries to airways that result from the transplantation surgery and cytomegalovirus infection. PMID- 10813205 TI - Algorithm for diagnosing pulmonary fibrosis in tropical countries. AB - An algorithm for diagnosis of pulmonary fibrosis in the tropical countries has been developed on the basis of the common causes of fibrosis, and the availability and feasibility of different diagnostic techniques in those countries. First, it is important to exclude common diseases such as the atypical or occult forms of bronchiectasis, pulmonary tuberculosis, and chronic bronchitis, which often overshadow interstitial pulmonary fibrosis. A good history and physical examination supplemented with chest radiography and simple lung function tests are generally enough to narrow down the list of causes of diffuse lung disease to interstitial pulmonary fibrosis. The real difficulty lies in identifying the idiopathic or "lone" forms from the secondary forms of pulmonary fibrosis. High-resolution CT is helpful in a large number of patients. Transbronchial lung biopsy is performed in a select population of patients. Open surgical or thoracoscopy guided biopsy is the gold standard, but is rarely required. PMID- 10813206 TI - Hyperoxia and lung disease. AB - Experimental studies and few human reports demonstrate that hyperoxia increases the level of reactive oxygen-derived free radicals and that these substances can produce oxidative cellular injury. However, available data suggest that the human lung is more resistant to hyperoxic oxidative damage than previously expected and demonstrate that absorption atelectasis is the most frequently pulmonary effect of inhalation of a high inspired oxygen fraction. Practically, the therapeutic use of high inspired oxygen fractions is limited to patients with acute lung injury and severe hypoxemia. Recent studies demonstrated that ventilator-induced lung injury is a more important cause of pulmonary damage in these patients than hyperoxic toxicity. New protective ventilatory strategies are associated with increased survival. PMID- 10813207 TI - Progressive systemic sclerosis and the lung. AB - Pulmonary manifestations of systemic sclerosis (scleroderma) are many. The mean survival in these patients is 78 +/- 17 months. Improved diagnostic techniques and better therapeutic options are essential to stem the tide of overwhelming mortality in patients with scleroderma lung disease. PMID- 10813208 TI - Management and care of the newly diagnosed patient with cystic fibrosis. AB - Cystic fibrosis is the most common autosomal recessive fatal disease among whites. Life expectancy is now at 31 years of age. The major cause of morbidity and mortality is chronic progressive lung disease. Lung disease occurs early in cystic fibrosis, suggesting the need for early and aggressive treatment of any pulmonary symptoms and malnutrition. This treatment is ideally performed within a cystic fibrosis center, and new therapies are being sought for management of the early stages of disease. PMID- 10813209 TI - Gastrointestinal surgery in cystic fibrosis. AB - This review is an attempt to summarize relevant gastrointestinal surgical issues in the patient with cystic fibrosis. Many of the surgical treatments are standard and have remained unchanged for several years and are only briefly discussed. A few areas with new developments are meconium ileus and the implications of prenatal diagnosis of meconium peritonitis. In addition, new findings with hepatobiliary disease and gastroesophageal reflux disease associated with cystic fibrosis patients may change the manner in which these entities are approached because these patients are now living longer. Finally, we review the recent findings associated with fibrosing colonopathy. PMID- 10813210 TI - Measurement of quality of life in cystic fibrosis. AB - Considerable progress has been made over the past 2 decades in defining and measuring health-related quality of life (QOL), and there is a growing recognition that these measures provide unique information about the impact of a chronic illness and its treatment. For patients with cystic fibrosis (CF), health related QOL measures enable researchers and clinicians to determine the effects of clinical interventions on several aspects of daily living (psychological, emotional, social) that are not reflected in typical health indicators, eg, pulmonary functioning scores. Three types of health-related QOL measures have been developed: 1) utility measures, 2) health profiles, and 3) disease-specific measures. The purpose of each type of health-related QOL measure is described, and its application to patients with CF is reviewed. Although important descriptive information has been obtained from utility measures, eg, the Quality of Well-Being Scale, and health profiles, eg, the Nottingham Health Profile, both of these instruments have serious limitations. Disease-specific measures, similar to those developed for children and adults with asthma, seem to hold the greatest promise for advancing our understanding of the impact of CF on daily life and for evaluating the effectiveness of new clinical interventions. The Cystic Fibrosis Questionnaire is the only published disease-specific measure of health-related QOL for children, adolescents, and adults with CF. Its psychometric properties are briefly reviewed, and directions for future research are suggested. PMID- 10813211 TI - End-of-life care in cystic fibrosis. AB - Despite significant progress in therapy for cystic fibrosis (CF), most patients with the disease still die before the age of 30 years. Published discussions and descriptions of end-of-life care of patients with CF have been few, but interest in these issues is increasing. Descriptions of end-of-life care for persons with CF have begun to appear, and conflicting models of care have been proposed. Advanced care planning offers an opportunity for persons with CF to actively influence the type of care they will receive. Still, more empiric research and ethical discussion is needed to facilitate optimal end-of-life care of patients with CF. PMID- 10813212 TI - Burkholderia cepacia epidemiology and pathogenesis: implications for infection control. AB - Much has been learned during the past 2 years about the microbiology and taxonomy of Burkholderia cepacia. Several distinct species have been identified in what is now referred to as the B. cepacia complex. Preliminary studies indicate that certain of these species are more likely to colonize and cause severe pulmonary infection in persons with cystic fibrosis. Ongoing investigations will expand these findings and have the potential to modify current infection control policies. The commercial use of B. cepacia as an antifungal biopesticide and in bioremediation has attracted increasing attention from industry recently and raises concerns within the cystic fibrosis community. Consensus regarding the potential threat of such uses to cystic fibrosis patients is being sought by governmental agencies and agricultural and biomedical researchers. PMID- 10813213 TI - The role of the upper airway in sleep. PMID- 10813214 TI - Surgical approaches to obstructive sleep apnea. AB - Obstructive sleep apnea results from a combination of a structurally small upper airway and loss of upper airway muscle tone. Surgical therapy is effective by enlarging the upper airway and decreasing collapsibility. Limited palatal surgery has been demonstrated successful for the treatment of snoring but less for the treatment of obstructive sleep apnea. More aggressive multilevel surgeries addressing several airway segments demonstrate improved success rates. Multilevel surgeries and combined with bimaxillary advancement surgery have demonstrated success rates of 90%. New surgical procedures, eg, radiofrequency volume reduction, offer the potential of altering the upper airway with low morbidity. Hypoglossal nerve stimulation is a potential innovative technique. PMID- 10813215 TI - Tolerance and intolerance to continuous positive airway pressure. AB - Nasal continuous positive airway pressure (CPAP) is the most effective and widely used therapy for obstructive sleep apnea. As with any chronic therapy, long-term compliance has a significant impact on its effectiveness. Only about half of patients use CPAP for more than half the night on five or more nights per week. Approximately 4 hours of CPAP therapy per night appears to significantly improve daytime alertness and performance. Four hours of therapy also seems to improve sleep-disordered breathing for the remainder of the night. Patient education and close follow-up and intervention appear to improve long-term tolerance. Autotitration CPAP or bilevel positive airway pressure systems are no more effective or better tolerated than conventional CPAP therapy. They may be useful options if patients have been unable to tolerate conventional CPAP therapy. PMID- 10813216 TI - Oral appliances in the treatment of obstructive sleep apnea and snoring. AB - Treatment of snoring and obstructive sleep apnea with oral appliances is a promising alternative to continuous positive airway pressure (CPAP) therapy. Although there is wide variability in appliance design, these devices produce enlargement of the upper airway by advancing the mandible, tongue, or both. Oral appliances effectively reduce snoring in a significant proportion of individuals. In general they are less effective than CPAP in eliminating obstructive sleep apnea, with approximately 50% of individuals experiencing resolution of their obstructive sleep apnea with these devices. Patients with milder disease have greater success with oral appliance therapy than those with more severe disease. A small proportion of patients will experience worsening of their sleep apnea with an oral appliance, and close follow-up after device prescription is mandatory. Patient satisfaction with oral appliances may be better than that with CPAP. PMID- 10813217 TI - The nose and obstructive sleep apnea. AB - The role of the nasal passage in the pathophysiology of obstructive sleep apnea (OSA) is still controversial. To this end, induction of acute nasal obstruction in healthy volunteers is associated with sleep fragmentation and an increase in the number of obstructive and central apneas. Moreover, nasal topical anesthesia, which alters nasal reflexes, is associated with an increase in the number of obstructive and central apneas during sleep. Although nasal resistance is increased in patients with OSA, chronic nasal obstruction appears to play no significant role in the genesis, maintenance, or severity of OSA. Moreover, surgical correction of nasal obstruction does not alleviate OSA appreciably. The purpose of this review is to outline the current body of knowledge on the pathophysiology, diagnosis, and treatment of nasal obstruction in patients with OSA. PMID- 10813218 TI - Mechanical properties of the upper airway. AB - Abnormalities of upper airway mechanical properties are a well-recognized and important feature of the pathophysiology of the obstructive sleep apnea hypopnea syndrome (OSAHS). Recently, investigations enhanced our understanding of the factors that promote upper airway obstruction. In patients with OSAHS, anatomic narrowing of the pharyngeal airway, particularly in the lateral dimension with thickening of the lateral pharyngeal walls, is present. In addition, the passive upper airway (absent muscle activity) demonstrates increased collapsibility, which is modulated by caudal tracheal traction, mucosal surface forces, route of breathing, and the balance of intraluminal airway and extraluminal tissue pressures. In patients with OSAHS, pharyngeal dilator muscles (including the genioglossus and soft palate muscles) demonstrate a coordinated pattern of increased muscle activity while awake compared with normals. This is thought to represent a neuromuscular compensatory mechanism for the anatomically narrow, more collapsible upper airway. With the onset of sleep, the reflexes that drive this muscular compensation are diminished, leading to reduced muscle activity and predisposing the OSAHS patient to pharyngeal collapse. Better understanding of the mechanical properties of the upper airway in normals and patients with OSAHS should help in the development of new therapeutic strategies. PMID- 10813219 TI - Electric stimulation of the upper airway muscle. AB - In various studies, it has been postulated that pharyngeal collapse occurring during sleep in obstructive sleep apnea may be alleviated by stimulating the genioglossus muscle. Basic experiments have demonstrated that neuromuscular stimulation applied intraorally via electrodes or by direct neural stimulation of the hypoglossal nerve might improve upper airway and respiratory function. An increase of maximal inspiratory airflow, an improvement of upper airway collapsibility and a decrease in respiratory events during sleep were observed. An impairment of sleep quality during electric stimulation has been excluded simultaneously. Considering clinical aspects, anatomical properties and long-term experience in electric stimulation it might be possible to develop full implantable devices as an alternative treatment for patients with obstructive sleep apnea. PMID- 10813220 TI - Upper airway reflexes. AB - It is usually assumed that upper airway pressure receptors mediate the reflexes involved in sleep apneas, but many other receptors may be involved, including those responding to chemical stimuli. The reflexes to upper airway negative pressure have been further studied, and the timing of their inputs shown to be important. Their effects on the cardiovascular system, including cerebral blood flow, have been emphasized. The central nervous pathways for the upper airway reflexes and their relationship to the neuronal circuits of the respiratory rhythm generator are being analyzed, but no clear pattern has emerged. Many neurotransmitters have been identified, usually on the motor pathways, which points to possible therapeutic approaches. The central nervous pharmacology and the neuronal pattern for the cough reflex have been described, and a similar approach to other upper airway reflexes, especially those involved in sleep apneas, would be valuable. PMID- 10813221 TI - The use of negative pressure ventilation in infants with acute respiratory failure: old technology, new idea. PMID- 10813222 TI - Complications of noninvasive ventilation. PMID- 10813223 TI - Initial experience with a respiratory therapist arterial line placement service. AB - BACKGROUND: Indwelling arterial lines are commonly used in critical care. To standardize and improve the placement of these devices, we developed and implemented a respiratory therapist-based line placement service. As a measure of the quality of the service, we assessed the success and complications encountered in the first 119 line placement attempts of this new service. METHODS: The following were recorded for each artery on which cannulation was attempted: the number of the attempt on which cannulation was successful; if a different person was able to cannulate the artery after initial failure; and whether any complications occurred. Success rate compared to the number of attempts was tested with chi-square. RESULTS: Respiratory therapists were successful in placing 80% of attempted lines on the first try, including all 18 of 18 dorsal pedis attempts. Ninety-seven percent (115 of 119) of attempted arteries were ultimately cannulated. Success on second attempts by the same person was less than if a different, more experienced, person attempted the placement (p = 0.024). No complications were identified during the study. CONCLUSIONS: Initiation of a respiratory therapist-based arterial line placement service resulted in an acceptable cannulation success rate, without complications. Increased experience of the person attempting cannulation correlates with improved success. PMID- 10813224 TI - Negative pressure ventilation via chest cuirass to decrease ventilator-associated complications in infants with acute respiratory failure: a case series. AB - Pulmonary and nonpulmonary complications of invasive positive pressure ventilation are well documented in the medical literature. Many of these complications may be minimized by the use of noninvasive ventilation. During various periods of medical history, negative pressure ventilation, a form of noninvasive ventilation, has been used successfully. We report the use of negative pressure ventilation with a chest cuirass to avoid or decrease the complications of invasive positive pressure ventilation in three critically ill infants at two institutions. In each of these cases, chest cuirass ventilation improved the patient's clinical condition and decreased the requirement for more invasive therapy. These cases illustrate the need for further clinical evaluation of the use of negative pressure ventilation utilizing a chest cuirass. PMID- 10813225 TI - Inspissated secretions: a life-threatening complication of prolonged noninvasive ventilation. AB - Noninvasive Ventilation is frequently initiated in an attempt to avoid the complications of invasive mechanical ventilation. The optimal duration of this strategy is unclear, as prolonged noninvasive ventilation has associated complications. This case report illustrates the development of life-threatening inspissated secretions precipitating airway obstruction as a consequence of prolonged noninvasive ventilation. PMID- 10813226 TI - The use of noninvasive ventilation in acute respiratory failure associated with oral contrast aspiration pneumonitis. AB - Noninvasive ventilation (NIV) has been used to treat patients with acute respiratory failure, including cases of pneumonia. We used this technique in the management of an 83-year-old patient with acute respiratory failure secondary to inadvertent administration of oral contrast material into the lung, and who did not want to be intubated. NIV resulted in immediate improvement of respiratory status. The patient was weaned from NIV over the next 24 hours and eventually discharged from the hospital. PMID- 10813227 TI - Bronchodilator resuscitation in the emergency department. Part 2 of 2: dosing strategies. PMID- 10813228 TI - Office spirometry for lung health assessment in adults: a consensus statement from the National Lung Health Education Program. AB - Chronic obstructive pulmonary disease (COPD) is easily detected in its preclinical phase using spirometry, and successful smoking cessation (a cost effective intervention) prevents further disease progression. This consensus statement recommends the widespread use of office spirometry by primary-care providers for patients > or = 45 years old who smoke cigarettes. Discussion of the spirometry results with current smokers should be accompanied by strong advice to quit smoking and referral to local smoking cessation resources. Spirometry also is recommended for patients with respiratory symptoms such as chronic cough, episodic wheezing, and exertional dyspnea in order to detect airways obstruction due to asthma or COPD. Although diagnostic-quality spirometry may be used to detect COPD, we recommend the development, validation, and implementation of a new type of spirometry-office spirometry--for this purpose in the primary-care setting. In order to encourage the widespread use of office spirometers, their specifications differ somewhat from those for diagnostic spirometers, allowing lower instrument cost, smaller size, less effort to perform the test, improved ease of calibration checks, and an improved quality-assurance program. PMID- 10813229 TI - What causes an elevated diffusing capacity? PMID- 10813230 TI - A 56-year-old woman with mixed obstructive and restrictive lung disease. PMID- 10813231 TI - Pancoast tumors of the lung. AB - Pancoast or superior pulmonary sulcus tumors are uncommon primary bronchogenic carcinomas that produce a characteristic clinical syndrome of upper extremity pain and Horner's syndrome. Treatment of patients with this malignancy has traditionally involved irradiation alone or preoperative irradiation followed by resection. Recent advances in the management of Pancoast tumors include the importance of mediastinoscopy in staging the tumor before treatment begins. A complete resection should be accomplished including a lobectomy whenever possible. The current treatment protocol involves induction chemoradiotherapy followed by surgery. PMID- 10813232 TI - Epidemiology of lung cancer. AB - Lung cancer incidence is now decreasing in US men. Although rates continue to increase in women, the rate of increase is declining. Most lung cancer in men and women is attributable to cigarette smoking. Histologic patterns are consistent with smoking trends for gender, race, and age. Trends in adenocarcinoma may be related to an increase in exposure to tobacco-specific nitrosamines from low-tar cigarettes. Other risk factors, including exposure to residential radon, occupational exposures, diet, and family history, have been shown to increase risk of lung cancer independent of cigarette smoking. Recent research in molecular epidemiology has greatly increased our understanding of the mechanism of lung carcinogenesis and the interactions between exposure to lung carcinogens (smoking, occupational exposures, radon), diet, and heritable variations in susceptibility. PMID- 10813233 TI - Photodynamic therapy for bronchogenic carcinoma. AB - Photodynamic therapy has been used in the management of early lung cancer since the 1980s. It appears to be effective as a curative therapy for superficial squamous cell carcinomas and as palliative therapy in obstructive cancers of the tracheobronchial tree. New photosensitizers that have a deeper penetration and less photosensitivity are needed. PMID- 10813234 TI - Molecular biology of lung cancer. AB - Lung cancer is a major cause of mortality worldwide and the overall survival rate has not improved significantly in the past 20 years. Of the more than 150,000 new cases of lung cancer diagnosed in Europe every year, less than 10% of patients can be cured and enjoy long-term survival. The role of tobacco smoking as a cause of lung cancer has been conclusively established. Early diagnosis of lung cancer, based on conventional screening procedures using sputum cytology and chest radiography, has been so far unable to decrease lung cancer mortality. We discuss here the possibility that novel, more specific molecular markers, beside providing new understanding of the process of lung carcinogenesis, may also constitute new tools for early diagnosis allowing for screening of high-risk individuals, determination of prognosis, and identification of innovative treatments. PMID- 10813235 TI - Disorders of pulmonary circulation. PMID- 10813236 TI - Anticoagulation management clinics for the outpatient control of oral anticoagulants. AB - Oral anticoagulant therapy has a high risk-benefit profile and is labor intensive to manage. A better understanding of the indications for anticoagulation and of prothrombin time monitoring has led to improved outcomes. The development of anticoagulation management services has also improved the overall management of anticoagulation and the resulting clinical outcomes. By reducing adverse events, anticoagulation management services also result in more cost-effective therapy. Although these results have been generated by mostly nonrandomized observational studies, until better quality investigations are available, the preponderance of current clinical evidence strongly supports the value of a coordinated approach to care by an anticoagulation management service. PMID- 10813237 TI - Low molecular weight heparin versus unfractionated heparin in the initial treatment of venous thromboembolism. AB - In this review, we analyze data from randomized trials in which low molecular weight heparin was compared with unfractionated heparin, both to estimate the treatment effect of low molecular weight heparin in the initial treatment of venous thromboembolism and to evaluate the effect of the varied proportion of included cancer patients (6% to 22.7%) on the incidence of outcome events (recurrence of venous thromboembolism, bleeding, and mortality) and on the estimated treatment effect. Low molecular weight heparin has been extensively investigated in patients with deep vein thrombosis, but few trials have included patients with pulmonary embolism. The risk of recurrence of venous thromboembolism (odds ratio, 0.77; 95% CI, 0.56-1.04), major bleeding (odds ratio, 0.60; 95% CI, 0.38-0.95), and mortality (odds ratio, 0.72; 95% CI, 0.55 0.96) was less with low molecular weight heparins compared with unfractionated heparin. The proportion of cancer patients in these studies had a statistically significant effect on the incidence of recurrent venous thromboembolism (P = 0.03) and mortality (P = 0.002), but no influence on the estimated treatment effects of low molecular weight heparins. Low molecular weight heparin is effective and safe in the initial treatment of venous thromboembolism. PMID- 10813238 TI - The management of parapneumonic effusions and empyema. PMID- 10813239 TI - Antibiotic treatment of patients with pneumonia and pleural effusion. AB - Despite all medical advances of this century, pneumonia is still a common condition and the sixth leading cause of mortality in the United States. A pleural effusion develops in up to 40% of cases of pneumonia and empyema in 5% to 10% of those patients. The morbidity and mortality rates in patients with pleural effusions as a consequence of pneumonia are higher than in patients with pneumonia alone. In this review we discuss the classification, bacteriology, and the appropriate antibiotic treatment of patients with parapneumonic pleural effusions. PMID- 10813240 TI - Medical thoracoscopy. AB - Thoracoscopy has received increasing attention over the past decade as a result of the considerable advances that have been made in the development of endoscopic instruments. In contrast to the newly established video-assisted thoracoscopic surgery, the classic way to perform thoracoscopy is using only local anesthesia and sedation (medical thoracoscopy) making the procedure less invasive and expensive. The leading diagnostic indication for medical thoracoscopy today is an exudative pleural effusion of unknown origin offering a yield of more than 90% in malignancy or tuberculous pleurisy. In addition, talc poudrage during thoracoscopy is the most effective way to perform pleurodesis. For spontaneous pneumothorax, the second most important indication, medical thoracoscopy allows staging as well as therapeutic measures such as coagulation of blebs or talc poudrage. Other indications such as biopsy for diffuse lung disease or peripheral nodules are now reserved for video-assisted thoracoscopic surgery. PMID- 10813241 TI - Hepatic hydrothorax. AB - Hepatic hydrothorax is the accumulation of ascitic fluid in the pleural space and requires the same treatment as ascites: salt restriction, diuretics, and paracentesis. Refractory hydrothorax appears when there is no response to those measures and its management is not well established. Videothoracoscopy is a promising therapy that permits the detection and closure of diaphragmatic defects, and when used with pleurodesis resulted in long-lasting control of hydrothorax in six of eight patients without appreciable morbidity. The transjugular intrahepatic portosystemic shunt is an effective therapy in more than 75% of refractory hydrothorax cases. Hepatic encephalopathy and worsening of liver function in some patients are the main adverse effects. Spontaneous bacterial empyema, the infection of a hydrothorax, was reported in 13% of 120 cirrhotic patients with hydrothorax. Forty percent of the episodes of spontaneous bacterial empyema were not associated with spontaneous bacterial peritonitis. The sensibility of pleural fluid culture improves inoculating pleural fluid into a blood culture bottle at the bedside. Patients with refractory hydrothorax or those having an episode of spontaneous bacterial empyema should be considered candidates for liver transplantation. PMID- 10813242 TI - Thoracoscopy in the management of hemothorax and retained blood after trauma. AB - Retained hemothorax complicated by blood clotting in the thoracic cavity traditionally has been treated with open thoracotomy for evacuation of the hemothorax and cleaning of the thoracic cavity. Recent improvements in video technology and endoscopic surgical instruments have fostered renewed interest in video-assisted thoracoscopic surgery to diagnose and treat a variety of surgical conditions of the chest, which classically were managed exclusively by thoracotomy. Posttraumatic retained hemothorax currently is being managed at most institutions by video-assisted thoracoscopic surgery, with consistently good results. PMID- 10813243 TI - Perinatal events in the development of asthma. AB - Many potential factors are likely involved in the pathogenesis of asthma. These include prenatal, peripartum, and postnatal influences. Prenatally, genetic endowment, maternal smoking, in utero allergen sensitization, and alterations in maternal immune function, especially at the placental level, may increase the risk for asthma and atopy. In the peripartum period, suspected factors include obstetric practices (eg, the use of prostaglandins, hormones, and other agents) and prematurity. Postnatally passive smoke exposure, neonatal or early childhood infections and breast-feeding are under increasing scrutiny as to their possible role in the development of asthma. Despite the volumes of work already reported, much more is left to be done to sort out the complex interrelationships of these and other as yet unsuspected influences on the development of asthma. PMID- 10813244 TI - Immune dysregulation as a cause for allergic asthma. AB - Allergic asthma is being increasingly understood as a disease caused by Th2 mediated immune responses to inhaled allergens. Most individuals fail to respond to allergens with a Th2 response, and thus, allergic asthma can be considered the result of an abnormally regulated or dysregulated immune response. The prevalence of asthma has risen precipitously in urbanized cultures, as contrasted with third world countries. This observation underlies the heightened efforts in the past few years of basic and applied research efforts to gain a better understanding of both normal and dysregulated immunity to antigens introduced via the airways. This review focuses on recent human studies into the immune dysregulation that results in the asthma phenotype, but also cites selected relevant papers from research with experimental animals. PMID- 10813245 TI - The 21st century environment and air quality influences on asthma. AB - "Criteria" air pollutants are federally regulated pollutants that occur widely outdoors and have diverse sources, most often related to combustion. They include ozone (O3), particulate matter, sulfur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (CO), and lead. All except lead may interfere with oxygen delivery, and so may be of special concern to asthmatics. In controlled experiments, SO2 causes acute asthma symptoms and bronchoconstriction, preventable by beta-agonist medications. Ozone causes acute irritant symptoms, restrictive lung dysfunction, increased bronchial reactivity, and lower-airway inflammation in healthy people and asthmatics. Exposures to O3, and possibly to other pollutants, appear to exacerbate bronchoconstrictive or inflammatory responses to inhaled aeroallergens (bioaerosols); this may represent an important health risk to asthmatics. Exposure levels known to evoke acute responses to O3 or SO2 are uncommon in community air pollution; however, some asthmatics might be susceptible to lesser, more common exposures. Evidence concerning NO2 is equivocal, but it may have O3-like effects in some asthmatics. Epidemiology has often associated particulate pollution with asthma exacerbations and other cardiorespiratory illnesses, even in cities with relatively mild air pollution. Current laboratory research cannot fully explain this association. Advances in emission controls should further reduce ambient pollution levels, but probably will not reduce asthma morbidity. Better asthma management, with improved anti inflammatory medications, more careful monitoring by patients and health care providers, and reasonable efforts to reduce pollutant and aeroallergen exposures, offers the best hope to reduce asthma morbidity in the new century. PMID- 10813246 TI - The challenge of inner-city asthma. AB - Racial and ethnic minorities of low socioeconomic status residing in urban environments currently referred to as inner cities appear to represent a population that is disproportionately at high risk for asthma morbidity and mortality. Epidemiologic studies suggest that key risk factors contributing to asthma morbidity within the inner city include social demography, the physical environment (indoor and outdoor), and health care access and quality. This epidemiologic literature has helped to define opportunities for successful intervention strategies in these high-risk populations. Studies of the effectiveness of community-based and health system-based interventions with specific focus on inner-city populations are beginning to emerge in the literature. PMID- 10813247 TI - The relationship between allergic rhinitis and bronchial asthma. AB - Allergic rhinitis and bronchial asthma frequently coexist in the same patients. Many patients with rhinitis alone demonstrate nonspecific bronchial hyperresponsiveness, and prospective studies suggest that nasal allergy may be a predisposing risk factor for developing asthma. A growing body of literature has documented that the treatment of patients with allergic rhinitis may result in improvement of asthma symptoms, airway caliber, and bronchial hyperresponsiveness to methacholine and exercise. Finally, mechanistic studies of airway physiology have demonstrated that nasal disease may influence pulmonary function via both direct and indirect mechanisms. Nasal disease should be considered as a possible concomitant illness in all patients with asthma and treated appropriately when identified. PMID- 10813248 TI - The role of cigarette smoke in the pathogenesis of asthma and as a trigger for acute symptoms. AB - Although it has been long believed that cigarette smoke is injurious to the lower respiratory tract, the exact early mechanisms and early events responsible for this injury remain unclear. Maternal smoking, particularly in utero, is clearly associated with an increased risk for the later development of childhood atopy and asthma. Smoking is known to increase the inflammatory burden of the lower respiratory tract through a number of related but separate mechanisms. These include the recruitment of increased numbers of inflammatory cells, alteration in cell subtypes, enhancement of some cellular functions, and proinflammatory mediator release. In addition, cigarette smoking in vitro and in animal models appears to promote neurogenic inflammation, increase oxidative stress and lead to the elevation of cysteinyl leukotrienes, all of which could potentially lead to an amplification of the airway inflammation already present in asthmatics. Greater and more consistent effort must be given to encourage the young asthmatic not to smoke. In addition, greater effort must be spent on smoking cessation, especially in pregnant women and young asthmatics. PMID- 10813249 TI - Case discussions on the pathophysiology and clinical features of near-fatal asthma episodes. AB - This article reviews the definition of near-fatal asthma. The slow-onset, late arrival group and the sudden-onset groups of near-fatal asthma patients are discussed. Risk factors for near-fatal asthma and the pathologic differences between the two groups are elucidated. PMID- 10813250 TI - Approach to the difficult pediatric asthmatic. AB - Wheezing in infants and children presents a difficult differential diagnosis contingent on the presenting symptoms and age of the child. A determination of the anatomy of the lower airway, combined with allergic, infectious, and noninfectious irritants, is requisite to a complete evaluation. The intervention strategies to decrease wheezing and bronchial hyperresponsiveness may include both nonpharmacologic and pharmacologic management. The nonpharmacologic management of asthma in children is contingent on aggressive treatment of dietary protein sensitivity and strict environmental control. The treatment of bronchial hyperresponsiveness with pharmacologic intervention in infants and children is relegated to the optimal use of sodium cromoglycate and inhaled corticosteroid. PMID- 10813251 TI - Management of the severe asthmatic. AB - Asthma morbidity and mortality continue to increase. The clinical characteristics of the high risk asthmatic patient continue to be elucidated. These include historical features, current disease characteristics and psychosocial factors. Beta-Adrenergic agonists continue to be the mainstay of acute therapy. The following review details these topics. PMID- 10813252 TI - Gastroesophageal reflux disorders and asthma. AB - Gastroesophageal reflux (GERD) may trigger asthma. Approximately 77% of asthmatic people experience reflux symptoms, although GERD may be clinically silent in some. Esophagitis is found in 43% of asthmatic people, and 82% have abnormal esophageal acid contact times on esophageal pH testing. Clearly, GERD is prevalent in asthmatic people. Pathophysiologic mechanisms of acid-induced bronchoconstriction include a vagally mediated reflex and microaspiration. Whether these airway responses are clinically significant is the subject of some debate. Interestingly, peak expiratory flow rates and specific airway resistance alterations persist despite esophageal acid clearance. Preliminary evidence shows that substance P, an inflammatory mediator that causes airway edema, is released with esophageal acid. Although therapeutic studies are limited by their small population sizes and study design, up to 70% of asthmatic people have asthma improvement with antireflux therapy. Possible predictors of asthma response include patients with symptomatic esophageal regurgitation; abnormal proximal esophageal acid exposure; and, in surgical studies, those with normal esophageal motility and asthma response with medical therapy. Future research will further define the association between asthma and gastroesophageal reflux. PMID- 10813253 TI - Physical activity and asthma. AB - The relationship between asthma and exercise and the resultant disability (ie, the impact on activities of daily living, including physical activity) shows wide interpatient and intrapatient variability, being influenced not only by the disease but additional psychosocial variables. There are a variety of helpful pharmacologic and nonpharmacologic measures in dealing with simple exercise induced asthma, and new therapeutic options are being developed. The cardiorespiratory performance characteristics of asthmatic patients are very frequently suboptimal, either because of symptom-limited exercise tolerance or secondary deconditioning consequent upon inactivity. Medically supervised physical training can produce significant beneficial change. Recommendations for rehabilitation of asthmatic patients would include individualized exercise prescription and advice based on objective criteria of exercise capability, with flexibility in the programs offered, in order to cater to the broadest spectrum of patient disability. PMID- 10813255 TI - Bibliography. Current world literature. Asthma. PMID- 10813254 TI - Anti-IgE as novel therapy for the treatment of asthma. AB - Immunoglobulin-E (IgE) is believed to be the central effector antibody reacting to allergen in patients with allergic asthma. Clinical manifestations of allergic asthma result from the release of chemical mediators from mast cells and basophils on exposure to allergen. A humanized murine monoclonal antibody to IgE, rhuMAb-E25, recognizes the specific Fc epsilon3 portion of circulating IgE that binds to the high-affinity IgE receptor, Fc epsilonRI. In clinical studies, single and multiple doses of subcutaneous and intravenous rhuMAb-E25 have been shown to reproducibly reduce the serum free IgE concentrations in a dose dependent manner. Clinical trials conducted in aeroallergen bronchoprovocation laboratories demonstrated that decreasing circulating IgE resulted in significant attenuation of the early and late asthmatic responses. Studies completed in moderate to severe allergic asthmatics have extended the safety and efficacy of rhuMAb-E25. Significant improvements in asthma symptoms, meaningful reductions in corticosteroid agents while decreasing reliance on bronchodilator rescue drugs, decreased asthma exacerbations, and improved quality of life have been documented. Because of the remarkable protein engineering and the humanization technology now available, rhuMAb-E25 therapy has elicited no antibody responses and has been safely administered to atopic subjects. rhuMAb-E25 as a novel monoclonal antibody, the first to be applied to lung diseases, holds promise for the control of many IgE-mediated diseases. PMID- 10813256 TI - Effects of long-term oxygen therapy in patients with chronic obstructive pulmonary disease. AB - Long-term oxygen therapy is largely used in the management of severe hypoxemia in patients with chronic obstructive pulmonary disease. It was demonstrated that long-term oxygen therapy prolongs life, prevents progression of hypoxic pulmonary hypertension, and controls polycythemia. Recent data suggest that in patients with moderate hypoxemia (Pao2 > 55 mm Hg), long-term oxygen therapy does not prolong life. Life expectancy in those patients seems to depend on the severity of airway obstruction. Long-term oxygen therapy improves cognitive functions and emotional status. There is some evidence suggesting that it also improves quality of life, but more data are needed. There are conflicting data concerning the rationale for nocturnal oxygen supplementation in patients with arterial blood desaturation during sleep. PMID- 10813257 TI - The role of theophylline in asthma management. AB - Views on the appropriate use of theophylline in asthma management have varied substantially over the past decades. The recent emphasis on potential anti inflammatory effects of theophylline has only added to the debate. In current guidelines, theophylline has been positioned mainly as a form of "add-on" therapy in moderate to severe persistent asthma. The purpose of this review is to analyze whether recent developments have been made that allow for a further positioning of theophylline in the treatment of asthma. PMID- 10813258 TI - Worldwide epidemiology of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) continues to cause a heavy health and economic burden around the world. Recent studies have added evidence to the etiologic role of known and suspected risk factors for lung function decline and COPD, including smoking, occupational exposures, air pollution, airway hyperresponsiveness, and certain genetic variations. Among most populations, COPD prevalence and mortality are still increasing and will likely continue to rise in response to increases in smoking, particularly by women and adolescents. Resources aimed at smoking cessation and prevention and the early detection of COPD will be of the most benefit in our continuing efforts against this important cause of morbidity and mortality. PMID- 10813259 TI - Economic consequences of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is an important preventable cause of morbidity and mortality throughout the world. Little is known, however, about the economic burden of COPD imposed on society. This paper reviews the medical and economic literature pertaining to COPD in order to provide information and perspective for clinicians and those responsible for making payment and resource allocation decisions. There are only two published estimates of the burden of illness of COPD. In 1970, the estimated societal burden of emphysema was US$1.5 billion. Average individual expenditures for persons with COPD belonging to Medicare health maintenance organizations have been estimated to be more than twice as high as similar individuals without COPD. The burden of COPD on society is expected to increase substantially over the next three decades. COPD is a unique public health challenge to society now and for the foreseeable future. PMID- 10813260 TI - Health-related quality of life in asthma. AB - One aim of caring for adults and children with asthma should be the identification and treatment of the functional impairments that are troublesome to these patients in their daily lives. Studies have shown that correlations between measures of clinical asthma severity and control and health-related quality of life (HRQL) impairment are only weak to moderate. Therefore, HRQL must be measured directly. In recent years, HRQL questionnaires have been developed and validated to measure the functional (physical, social, emotional, and occupational) impairments that are important to both adults and children with asthma. Most questionnaires are now available in a range of languages. More recently, methods have been developed for the clinical interpretation on HRQL data. Assessment of asthma-specific HRQL can be included in both clinical trials and clinical practice, in conjunction with the conventional measures of airway function, to provide a complete picture of patients' health status. PMID- 10813261 TI - Serum measurement of eosinophil cationic protein in the management of asthma. AB - Asthma is a disease characterized by chronic eosinophilic inflammation of the airways. Serum eosinophil cationic protein (ECP) has been increasingly used as a noninvasive inflammatory marker in asthma. The serum ECP level seems to reflect, although indirectly, the intensity of ongoing eosinophilic inflammation of the airways and respond sensitively to intervention, whereas it is unlikely to be useful for establishing the diagnosis of asthma in an individual patient. Monitoring of serum ECP could be of utility in the long-term follow-up of asthmatic patients. However, further longitudinal studies are required to establish the role of serum ECP measurement in the treatment modulation in asthma. PMID- 10813263 TI - Bibliography. Current world literature. Obstructive, occupational, and environmental diseases. PMID- 10813262 TI - Role of reactive oxygen species in occupational and environmental obstructive pulmonary diseases. AB - Free radicals and their metabolites, also called reactive oxygen species (ROS), have been implicated in the pathogenesis of many diseases. Because of its continuous exposure to toxic pollutants in the ambient air, such as cigarette smoke, air pollution, and mineral dusts, the lung is very vulnerable to ROS induced injury. In this review, the role of ROS in the pathogenesis of obstructive lung diseases is reviewed. A central theme in this review is the pivotal role of transition metals such as iron, vanadium, and nickel in ROS induced cell damage, not only in exposure to mineral dusts but also in cigarette smoke and air pollution. PMID- 10813264 TI - Police reports on domestic incidents involving intimate partners: injuries and medical help-seeking. AB - This study aims to expand knowledge of partner abuse injury and help-seeking through a new source of data. We reviewed police reports during corresponding periods in 1996 and 1997 (n = 476). Complainants were injured in 17.4% of all incidents; 90% of those injured were women. Hispanics were more likely to be injured than non-Hispanics (OR = 2.06; 95% CI = 1.0, 4.25). The likelihood of injury decreased with each year of increasing age (OR = 0.95; CI = 0.91, 0.99). Only 20.5% of those injured consented to medical care. Police reports provide information on partner abuse injury that supplements hospital surveillance and household surveys. PMID- 10813265 TI - Factors influencing participation in weekly support groups among women completing an HIV/STD intervention program. AB - Over the past three decades, the influence and importance of social support has been well documented and the findings have suggested a beneficial effect on stress-related situations, mental and physical health, and social functioning. More recently, small group/skills training behavioral interventions have demonstrated success in changing behaviors which affect the transmission of sexually transmitted diseases, including HIV among populations at risk for these diseases. Studies of support groups to date have been conducted exclusively in research settings where women are offered financial incentives for participation. Little is known about the willingness of women to participate in ongoing support groups after successfully completing a skills training intervention. The present study examines the factors that may influence participation among women in a weekly support group after completing a structured, six session HIV/STD intervention. Both quantitative and qualitative data are collected from 265 women in the intervention arm of a multi-site randomized controlled behavioral intervention trial. Results reveal that less than a quarter (22%) of women participated in at least one support group. Participation varied significantly by site, ranging from 34% to 15% (p = .008). Participation was also strongly linked to recent use of domestic violence services. Qualitative data indicated that although monetary incentives play some role in the woman's decision to participate, other factors are also important. These include program outreach, support group size, salience of the group content, consistency of group leadership from the intervention to the support group, and use of peer leaders along with professional facilitators. Implications for design of post intervention support groups programs are discussed. PMID- 10813266 TI - Use of health care services by women who use crack cocaine. AB - Chronic drug users demonstrate a need for access to health care due to both acute health problems related directly to substance use and to other existing medical problems. This study attempts to examine how women differ from men in their utilization of health services. Also, it analyzes how crack use affects men and women differentially. The study population is a community-based sample of 624, comprised equally of men and women, as well as crack users and non-users of crack. Results indicate that women utilized health care more than men; however, crack use among women appears to be an inhibitory factor in the utilization of health services by women. PMID- 10813267 TI - Facilitators and barriers to use of the female condom: qualitative interviews with women of diverse ethnicity. AB - Women in the United States, particularly African-Americans and Hispanics, are at increased risk for HIV. The female condom now offers women a potentially important option for HIV prevention, yet few efforts have been made to increase its use. To elucidate strategies to promote the use of the female condom, we conducted in-depth interviews with 62 women recruited from the four major racial/ethnic groups of the U.S. (African-American, Asian-American, Hispanic, and white). Subject recruitment took place at a family planning clinic in San Francisco during 1996-97. We identified four major types of facilitators and barriers to use of the female condom: mechanical, psychosexual, interpersonal, and situational. Specifically, the mechanical facilitators and barriers included positive and negative aspects of the device, and difficulty with insertion. The psychosexual factors were female empowerment, more options for contraception and disease prevention, discomfort with vaginal insertion, and condom use norms. The interpersonal factors included: enhanced communication, relationship status, partner preferences, and partner objections. Finally, the situations that made women disinclined to use the device were: no access to the female condom when having sex and using other forms of contraceptives. The implications of these findings for HIV prevention and future research are discussed. PMID- 10813268 TI - Symptoms of eating disorders among female distance runners: can the inconsistencies be unraveled? AB - Research on eating disorders among female distance runners has produced a modest, but inconsistent body of findings. To unravel the confusion, we hypothesized a model whereby studies finding greater symptomatology have involved obligatory runners or elite national/international competitors. Studies not finding greater symptomatology have involved a more typical group of athletes. To test our hypothesis, we used the Eating Disorders Inventory-2 (EDI-2) to compare 20 typical female collegiate distance runners to 35 female undergraduate psychology students. Comparisons were also made with norms in the EDI-2 Manual. Consistent with our model, the distance runners showed no enhanced symptomatology. Indeed, they seemed to be comparatively healthy. Operational definitions to further test the model are discussed. PMID- 10813269 TI - When the baby falls!: the cultural construction of miscarriage among Hmong women in Australia. AB - In this paper, traditional Hmong explanations about miscarriage and the ethnomedical knowledge and practices which pertain to it are explored. They are derived from in-depth interviews and participant observation with the Hmong who are now living in Melbourne, Australia. The loss of pregnancy creates considerable anxiety in Hmong society. This is not only because it calls for a socially justifiable explanation for a family's failure to extend their lineage, but also reduces the venue for a soul to be re-born into the family. This is a threat for Hmong society since it means the extinction of the family, clan and lineage and hence Hmong society. The cultural construction of the causes of miscarriage among the Hmong surrounds two main categories: the natural world which is related to the woman's body and her behaviour; and the supernatural world. These explanations point to the influence that individuals, both living and dead, have on pregnancies. In traditional Hmong society, health is perceived as a harmony between the social, and religious or supernatural realm. A woman being unable to bring forth an offspring because of miscarriage indicates disharmony with the living and/or between the living and the dead. PMID- 10813270 TI - "Remnants of feudalism"? Women's health and their utilization of health services in rural China. AB - Almost five decades ago, the Chinese Communist Party wished to abolish all "remnants of feudalism," including the patriarchal social order. Just one year after the revolution, the Marriage Law endorsed women's rights within the family, but no operative measures were taken to enforce it. Some of the economic reforms since independence even strengthened patrilocality and, possibly, patriarchal values. The purpose of this study was to explore the degree to which patrilocality served to maintain the traditional patriarchal stratification among women in the household by exploring women's health patterns and utilization of health services. Data were collected from 3859 women residing in rural Hebei, and variation in health and help seeking of six categories of relation to household head--mothers, wives, daughters, daughters-in-law, family heads, and other relatives--were explored. Utilization of health services is not dependent on women's position in the household, but primarily on per-capita income. Health patterns seem to indicate that mothers of the head of the household still have a considerable power to define their roles and share of household work. Women head of family, most of whom are married, appear to be under strain, which could be a result of their culturally "deviant" position. We conclude that old patriarchal values are intertwined with values of equality in current rural China. PMID- 10813271 TI - The epidemiology of psoriatic arthritis: fact or fiction? PMID- 10813272 TI - HLA-B27 misfolding and spondyloarthropathies: not so groovy after all? PMID- 10813273 TI - HLA-B27 polymorphism and association with disease. PMID- 10813274 TI - For example is not evidence: fibromyalgia and the law. PMID- 10813275 TI - Effects of methotrexate on the oxidative metabolism of cultured rabbit articular chondrocytes. AB - OBJECTIVE: The mechanism of action of methotrexate (MTX) in inflammatory joint disease is still unclear. We examined the possible interactions of MTX with the oxidative metabolism of rabbit articular chondrocytes. METHODS: Cell cultures of articular chondrocytes enzymatically isolated from juvenile New Zealand white rabbits were incubated 24 h with either MTX (0.22 or 1.1 microM), bacterial lipopolysaccharide (LPS, 50 microg/ml), or both. Cytofluorometry was then performed using 2',7'-dichlorofluorescein diacetate (DCFH-DA), rhodamine 123 (Rh123), or propidium iodide (PI). These fluorochromes allow evaluation of cellular production of H2O2, mitochondrial membrane potential, and cell viability, respectively. In a separate experiment, we used the Griess colorimetric technique to evaluate cellular nitric oxide (NO) production. RESULTS: Addition of MTX alone (0.22 or 1.1 microM) inhibited spontaneous production by chondrocytes of H2O2 (p < 0.01 and p < 0.001, respectively) and NO (p < 0.01 both concentrations). The LPS induced increase in H2O2, production was inhibited by MTX at 0.22 and 1.1 microM (p < 0.01 both concentrations), whereas the LPS induced increase in NO synthesis was not influenced by MTX, even at 1.1 microM. MTX did not significantly modify mitochondrial activity or cell viability. CONCLUSION: MTX at therapeutic concentrations in vitro inhibits the production of H2O2 and NO by unstimulated chondrocytes, and only the H2O2 overproduction by LPS stimulated chondrocytes. These properties may contribute to the therapeutic effect of MTX in RA. PMID- 10813276 TI - A histochemical study of the rheumatoid synovium: focus on nitric oxide, nerve growth factor high affinity receptor, and innervation. AB - OBJECTIVE: The synovium of patients with rheumatoid arthritis (RA) is characterized by massive cell proliferation, neoangiogenesis, and apoptosis. The nature of potential repressors/inducers of these phenomena is still largely unknown. We investigate if nitric oxide (NO) and nerve growth factor (NGF) can be considered potential mediators in these phenomena in RA. METHODS: Synovium of 15 patients with RA in active phase and synovium of 14 patients without synovial inflammation were processed for histochemical (NADPH-diaphorase) and immunohistochemical visualization of different isoforms for the NO synthesis enzyme NO synthase (NOS) and for NGF high affinity receptor trkA. RESULTS: Inducible NOS (iNOS) immunoreactivity and NADPH-diaphorase positivity were found in synoviocytes, fibroblast-like synoviocytes, fibroblasts, and inflammatory cells in the rheumatoid synovium. In the same areas and in the same cell types, although not in the same cells, we also found positivity for the NGF high affinity receptor trkA. CONCLUSION: We suggest that all elements involved in the transduction pathway that is activated by NGF and that proceeds through NO and tumor suppressor p53 are present in the synovium during RA, controlling cell cycle arrest, cell differentiation, and apoptosis. PMID- 10813277 TI - Analysis of Th1 and Th2 cytokines expressing CD4+ and CD8+ T cells in rheumatoid arthritis by flow cytometry. AB - OBJECTIVE: A Th1/Th2 cytokine imbalance with a predominance of Th1 cytokines has been suggested to be of pathogenetic importance in rheumatoid arthritis (RA). To evaluate the role of Th1/Th2 cytokines in RA, we used intracellular cytokine flow cytometry to determine cytokine profiles of CD4+ and CD8+ T cells in 34 peripheral blood (PB) and 10 synovial fluid (SF) samples from patients with RA. Results were compared with 10 PB samples from healthy controls (HC) and 5 SF samples from patients with non-RA synovitis. METHODS: After stimulating cells with PMA and ionomycin or alternatively with anti-CD3/CD28 in the presence of brefeldin A, intracellular levels of Th1 [interleukin 2 (IL-2), interferon-gamma (IFN-gamma)] and Th2 cytokines (IL-4, IL-5, IL-10, IL-13) were determined for CD3+CD8- (i.e., CD4+ Th1 and Th2 cells) and CD3+CD8+ (i.e., CD8+ Tc1 and Tc2 cells) T cells. RESULTS: The percentages of CD4+ and CD8+ Th1 and Th2 cytokines producing T cells (PB) were similar in patients with RA and healthy controls (HC), with a clear predominance of Th1 cytokines expressing, T cells. With regard to T cell subsets, IFN-gamma-producing T cells were significantly more frequently detected in the CD8+ subset [CD8+: median 45.1% (RA; p < 0.001), 38.2% (HC; p = 0.009) vs CD4+: 10.8%(RA), 17.0% (HC)]. Conversely, IL-2 was found in a higher percentage of CD4+ T cells [CD4+: median 33.4% (RA), 17.9% (HC) vs CD8+: 23.6% (RA), 12.3% (HC)]. Patients not in disease remission tended to have more IFN gamma-producing CD8+ and IL-2-producing CD4+ T cells than patients in remission [CD8+: median 45.9% (IFN-gamma) vs 23.0% (IFN-gamma); CD4+: median 34.1% (IL-2) vs 18.2% (IL-2)1. In all PB samples, the proportion of T cells producing the Th2 cytokines IL-4, IL-5, IL-10, and IL-13 did not exceed 2%. Cytokine profiles did not differ between patients receiving immunosuppressive treatment and patients treated only with nonsteroidal antiinflammatory drugs. In comparison to PB, RA SF analysis revealed a significant increase in the percentage of IFN-gamma-producing CD4+ (p < 0.001) and CD8+ T cells (p < 0.001). In addition, the percentage of IL 10-producing CD4+ (p < 0.001) as well as CD8+ T cells (p = 0.001) was significantly elevated in SF. However, production of the other Th2 cytokines (IL 4, IL-5, IL-13) was similar in SF and PB. CONCLUSION: These data indicate similar cytokine profiles of T cells in PB of RA patients and healthy controls, with a strong predominance of Th1 cytokines producing T cells in the CD4+ and CD8+ T cell subset of both groups. PB cytokine profiles did not significantly differ in patients with active and non-active disease or between patients receiving and those not receiving immunosuppressive medication. In SF, the proportion of Th1 and Tcl cells was significantly elevated compared to PB, emphasizing the local importance of these cells for inflammation. CD8+ T cells (Tc1 cells) mainly contributed to the production of IFN-gamma, indicating an underestimated role of this cell subset for local cytokine production. The upregulation of IL-10 producing Th2 and Tc2 cells in SF may reflect an insufficient effort to down regulate chronic inflammation in the joint. Modifying this cytokine imbalance in the joints may be a promising therapeutic approach in RA. PMID- 10813278 TI - Serum C-reactive protein does not predict rheumatoid arthritis. AB - OBJECTIVE: Clinical rheumatoid arthritis (RA) is preceded by an immunological process. Our objective was to investigate C-reactive protein (CRP) for its prediction of RA. METHODS: A case-control study was nested within a Finnish cohort of 19,072 adults who had no arthritis and no history of the disease at the baseline examination conducted between 1973 and 1977. By late 1989, 124 had developed RA. Three controls for each incident case were individually matched for sex, age, and municipality. The CRP concentration was measured by a sensitive ELISA. RESULTS: No difference was noted in serum CRP levels between cases and controls. The odds ratios (95% confidence intervals) of RA in the quintiles of CRP were 1.0, 0.89 (0.45-1.76), 1.11 (0.59-2.08), 0.99 (0.51-1.91), and 0.95 (0.49-1.87). Similarly, there was no difference when the data were analyzed in strata of sex, age, rheumatoid factor (RF) status at baseline, and length of time from taking the specimens to the onset of clinical disease. CONCLUSION: Pre rheumatoid immunological process as reflected in RF production is not associated with any marked inflammation or tissue injury heightening CRP. PMID- 10813279 TI - Factors influencing length of time taking methotrexate in rheumatoid arthritis. AB - OBJECTIVE: Duration of therapy has been suggested to represent a measure of effectiveness. Life table analyses of therapy with methotrexate (MTX) in rheumatoid arthritis (RA) have indicated a longer duration than with other drugs. However, individual patients continue taking MTX for different periods of time. We assessed the influence of patient variables at treatment onset upon subsequent duration of MTX therapy. METHODS: Patients with RA (n = 437) from 8 North American databank centers beginning MTX therapy after January 1, 1988, were followed prospectively. Age at onset of MTX treatment, sex, years of education, age at onset of disease, years with disease, number of comorbid conditions, number of disease modifying antirheumatic drugs (DMARD) and nonsteroidal antiinflammatory drugs (NSAID) taken just prior to MTX. disability level, pain, and global assessment prior to starting MTX were used in univariate Kaplan-Meier analyses to predict number of months taking MTX alone. An index that divided the patients into risk strata for predicting duration of therapy was constructed to be clinically useful. RESULTS: The median number of months continuing MTX without addition of other DMARD was 41 months and the median for the total course taking MTX was 52 months. The retention rate was lowest for patients with the most negative initial health state. High level of initial pain, long duration of disease, and not using a DMARD just prior to MTX were associated with low retention rate and can be used to predict expected durations of MTX treatment ranging from 17 to 52 months. For practical guidance in clinical decisions an index was computed based on the predictor variables: level of initial pain, duration of disease, and number of DMARD; this index identifies subgroups with very different durations taking MTX alone. Disease duration at baseline was strongly related to time taking MTX alone and could therefore also be used as a simplified rule in clinical work. CONCLUSION: Expected duration of MTX treatment is influenced by clinical variables, and these may suggest those patients likely to have more or less satisfactory experiences with MTX. The time taking drug alone (therapeutic segment) may be a more logical and sensitive indicator of effectiveness than the total course on the medication. PMID- 10813280 TI - Radiological and clinical results of longterm treatment of rheumatoid arthritis with methotrexate and azathioprine. AB - OBJECTIVE: To study whether the reported superior effect of methotrexate (MTX) compared to azathioprine (AZA) in retarding radiologic progression after one year in rheumatoid arthritis was sustained at 2 and 4 years. METHODS: All 64 patients enrolled in the original randomized double blind study were invited for an open extension of followup to 4 years including 4-monthly clinical and laboratory assessments and radiographs of hands, wrists, and feet at 2 and 4 years. RESULTS: After 4 years, 18 patients (58%) from the MTX group and 7 patients (21%) from the AZA group continued the initial study drug. During followup more patients (n = 21) switched from AZA to MTX than vice versa (n = 5). In an intention-to-treat analysis improvement of clinical and laboratory variables at 4 years was more pronounced in the MTX group. Mean radiologic scores increased in both treatment groups during followup. According to an intention-to-treat analysis increase in erosion score at one and 2 years in the MTX group was significantly lower than in the AZA group: after one year MTX group 1.8 versus AZA group 5.3 (p = 0.002); after 2 years MTX 3.5 versus AZA 6.5 (p = 0.05). After 4 years there was a trend toward less progression in the MTX group: MTX 6.8 versus AZA 10.8 (p = 0.09). For the total score, progression in the MTX group was less after one and 2 years. After 4 years marked radiologic progression was observed more often in the AZA group. CONCLUSION: Drug continuation after 4 years of followup was better for MTX than for AZA. In an intention-to-treat analysis the beneficial effect of MTX on radiologic progression compared with AZA was sustained after 2 years of followup. Thereafter differences between treatment groups leveled off, probably mainly due to the greater number of switches from AZA to MTX than vice versa. PMID- 10813281 TI - Direct and indirect costs associated with the onset of seropositive rheumatoid arthritis. Western Consortium of Practicing Rheumatologists. AB - OBJECTIVE: To examine the direct and indirect costs of rheumatoid arthritis (RA) during the first year of disease. METHODS: As part of a longitudinal observational study, 150 patients with seropositive RA of 5.9 +/- 2.9 mo duration were recruited through the Western Consortium of Practicing Rheumatologists. Subjects completed questionnaires about health care services and resources utilized and about the number of days of usual activity lost as a result of RA during the 6 month period prior to enrolment. RESULTS: Study participants had active RA as evidenced by mean tender and swollen joint counts of 24.9 +/- 13.5 and 20.6 +/- 11.6, respectively, and moderate functional impairment reflected by a mean Health Assessment Questionnaire (HAQ) score of 1.24 +/- 0.7. The average total direct cost of RA was $200/month. Health care visits, medications, and radiographs accounted for 78% of the total direct cost, while expenditures for hospitalizations accounted for only 3.5% of the total. The average number of days of usual activity lost per month because of RA was 3.8 +/- 7.7, translating into an average indirect cost of $281/month. Of the 95 subjects who were gainfully employed prior to disease onset, 12 were disabled and 5 were on sick leave as a result of RA, corresponding to a work disability rate of 18%. Work disabled subjects reported significantly lower total household incomes and higher HAQ disability and global disease activity scores than subjects who continued working. CONCLUSION: In this group of patients with seropositive RA substantial costs, both direct and indirect, were incurred during the first year of disease. PMID- 10813282 TI - KL-6 as a novel serum marker for interstitial pneumonia associated with collagen diseases. AB - OBJECTIVE: To investigate the diagnostic value of the serum concentrations of KL 6, a mucinous glycoprotein expressed on type II pneumocytes, for interstitial pneumonia (IP) in various collagen diseases. METHODS: Serum KL-6 levels were measured by ELISA. RESULTS: The mean values and the positive rates of serum KL-6 levels for patients with rheumatoid arthritis, systemic sclerosis, or polymyositis/dermatomyositis with IP were significantly higher than those without IP. Sensitivity, specificity, positive and negative predictive values of serum KL 6 level for IP associated with collagen diseases were 60.7, 98.9, 97.4, and 77.1%, respectively. The mean serum KL-6 level of patients with active IP was significantly (p = 0.0001) higher than that of patients with inactive IP. Serum KL-6 levels increased with the deterioration of IP, while the successful treatment of IP resulted in a significant decrease of these levels. CONCLUSION: Serum KL-6 concentration levels are a useful marker for diagnosis and evaluation of the disease activity of IP associated with collagen diseases. PMID- 10813283 TI - Rheumatoid arthritis in Sweden. Drug prescriptions, costs, and adverse drug reactions. AB - OBJECTIVE: Many patients with rheumatoid arthritis (RA) need continuous medication, bringing considerable costs for drugs and a need for monitoring adverse drug reactions. We studied drug exposure, drug costs, and adverse drug reactions in patients with RA in Sweden from 1987 to 1997. METHODS: Prescription patterns, drug costs, and adverse drug reactions, and their distributions and trends were analyzed by cross sectional annual data. Drug exposures and costs were assessed from the National Diagnosis and Therapy Survey. All costs were recalculated to the 1997 level using the drug price index. Information on adverse drug reactions was obtained from the national pharmacovigilance system. RESULTS: The drug prescription level was, on average, 2 defined daily doses (DDD) throughout the study period. Nonsteroidal antiinflammatory drugs (NSAID) accounted for > 40% of the drugs prescribed, methotrexate (MTX) 10%, corticosteroids 10%, and sulfasalazine 5%. Analgesics and opioids made up 14% of prescriptions - a low estimate considering the availability of over-the-counter preparations. Disease modifying antirheumatic drugs (DMARD) increased their proportion from 28.5 to 39.3%. Total costs were stable at $16 million US annually. NSAID costs decreased, while those of sulfasalazine, MTX, and cyclosporine increased. On average, 91 adverse drug reactions were reported annually. Hematological reactions (agranulocytosis, thrombocytopenia, leukopenia, pancytopenia) predominated, constituting 21% of reported reactions. Skin and gastrointestinal reactions, mainly mild, accounted for 15 and 14%, respectively. Deaths from adverse drug reactions were uncommon, about 3 per year, and were mainly attributed to hematological reactions. CONCLUSION: The total volume as well as the total cost of drug consumption for RA in Sweden was relatively constant during our 11 year observation period, despite a notable increase in the use of DMARD. This was mainly due to a decrease in costs per DDD of NSAID, and an increased use of cyclosporine. Drug related adverse reactions were dominated by hematological reactions, and fatal events were few. This emphasizes the need for an extended evaluation of which safety procedures are most cost effective for monitoring the drugs used for RA. PMID- 10813284 TI - Craniocervical junction involvement in rheumatoid arthritis: a clinical and radiological study. AB - OBJECTIVE: To make a comparative evaluation of different imaging techniques for studying the craniocervical junction involvement in patients with rheumatoid arthritis (RA). Upper cervical spine involvement was compared with clinical and immunological data. METHODS: Patients (n = 47) underwent plain radiographs and computerized tomography (CT) and magnetic resonance (MR) study of the craniocervical junction. Neurological examination following clinical signs of possible atlantoaxial involvement was performed in all patients following the Ranawat classification. RESULTS: Radiographic and MR images showed craniocervical involvement in 41.3% and 61% of the patients, respectively. Immunological data were not correlated with imaging findings, whereas Ranawat class II and III of neurological involvement seem to be predictive of atlantoaxial alteration. CONCLUSION: Conventional radiography allowed us to detect 41.3% of patients with craniocervical involvement, but only in advanced stages of the disease. MR imaging had the unique potential of direct and detailed synovial visualization, especially in the gadolinium enhanced axial images, resulting in the early diagnosis of craniocervical RA. PMID- 10813285 TI - Disease associated time consumption in early rheumatoid arthritis. AB - OBJECTIVE: To quantify the disease associated time consumption of normal activities of daily living and of treatment and monitoring activities in a cohort of patients with early rheumatoid arthritis (RA) with followup of at least 6 years. Comparison was made with a group of patients with asthma and chronic obstructive pulmonary disease (COPD). METHODS: A prospective and retrospective inventory was carried out, by interview and record investigation, of RA related and RA unrelated items covering the period from the start of the disease. Interviews were also performed in a group of patients with asthma and COPD. RESULTS: For patients with RA there was a mean disease associated time consumption of at least 1.9 h/day during the first 6 years of the disease. The time consumption was mainly due to extra time needed for activities of daily living and daily disease related activities. Patients with the greatest progression of radiographic damage, with the most severe disability, and with the greatest cumulative disease activity had the greatest time consumption. For patients with asthma and COPD the consumption of time was comparable. CONCLUSION: RA is a time consuming disease. Recognition of the disease associated time consumption will have implications for work (dis)ability assessments in patients with chronic diseases such as RA. PMID- 10813286 TI - Fibrinogen in systemic lupus erythematosus: more than an acute phase reactant? AB - OBJECTIVE: To investigate whether plasma fibrinogen (FNG) measured longitudinally in a cohort of patients with systemic lupus erythematosus (SLE) increased over observational time faster than in a control group, and whether its increase might depend upon age, disease duration, disease activity, and medications. METHODS: Hospital based retrospective study with repeated measurements of plasma FNG and C reactive protein (CRP) for patients and controls and erythrocyte sedimentation rate (ESR) and lupus activity index (LAI) for patients only. Study groups included patients with SLE: n = 96 (95% female), and healthy controls: n = 39 (95% female). Of the patients, 42% had SLE only, 23% had SLE with antiphospholipid antibodies (aPL), and 34% had SLE with aPL related thrombosis. RESULTS: Median baseline FNG was higher in patients (357 mg/dl; 95% CI 339-375) than in controls (271 mg/dl; 95% CI 251-291) by 86 mg/dl (95% CI 56-115, p < 0.001); in older subjects than younger (in patients and in controls); in patients with thrombosis than in other patient groups (by an average of 35 mg/dl; 95% CI 9 61 mg/dl): and in patients with longer disease duration (p = 0.05). Mean FNG increased faster in patients (19 mg/dl/year; 95% CI 12-26 mg/dl) than in controls (2.6 mg/dl/year; 95% CI 2.0-3.2 mg/dl). The increase was faster than the age effect and independent of patient group and disease activity. CONCLUSION: Plasma FNG in patients with SLE increases throughout followup regardless of disease activity, mimicking the age related increment observed in population based studies. The rapidity of the increment may reflect the prematurity of vascular disease typical of SLE. PMID- 10813287 TI - Studies of HLA-DR and DQ alleles in systemic sclerosis patients with autoantibodies to RNA polymerases and U3-RNP (fibrillarin). AB - OBJECTIVE: To determine the clinical and immunogenetic features of systemic sclerosis (SSc) patients with anti-RNA polymerase (RNAP) or anti-fibrillarin antibodies. METHODS: DNA typing for HLA-DR and DQ alleles was performed in 292 patients with SSc, including 81 with anti-RNAP and 24 with anti-fibrillarin antibodies. The remaining patients had anti-topoisomerase I (anti-topo I; 71), anti-centromere (ACA; 56), anti-Th/To (28), or other antinuclear (32) antibodies. RESULTS: Significant associations were observed in the patients with anti-topo I, ACA, and anti-Th/To antibodies, similar to those previously reported. No significant HLA associations were detected in the 81 patients with anti-RNAP. although weak associations were noted when this group was subdivided on the basis of immunofluorescence staining pattern; i.e., HLA-DR4 was increased in patients with strong nucleolar staining and HLA-DR3 was more frequent in patients with nucleoplasm staining only. No HLA-DR or DQ associations were observed in 24 patients with anti-fibrillarin antibodies. CONCLUSION: The identification of HLA associations in SSc patients with anti-RNAP antibodies may only be possible when the individual antibody specificities recognized by these sera are identified. It may then be possible to classify these patients into distinct clinical and immunogenetic subgroups. PMID- 10813288 TI - Autonomic nervous system and smooth muscle cell involvement in systemic sclerosis: ultrastructural study of 3 cases. AB - OBJECTIVE: To investigate morphological abnormalities in nerve and smooth muscle structures of the anorectal wall underlying gastrointestinal dysfunction in patients with systemic sclerosis (SSc). METHODS: We performed deep rectal biopsy in 3 patients with limited scleroderma of relatively recent onset and intestinal symptoms. RESULTS: We found ultrastructural signs of axonal degeneration and cytoskeletal abnormalities in the bundles of unmyelinated fibers. There was also focal degeneration of smooth muscle cells, often in association with the presence of partially degranulated mast cells. Many mast cells were also observed in close relation to nerve fibers and vessels. The enteric vessels often showed basal lamina reduplication and hypertrophied endothelial cells with obliterated lumen. No significant fibrosis was found. CONCLUSION: Our findings indicate early involvement of the autonomic nervous system and to a lesser extent of smooth muscle cells. We confirmed the presence of early vascular lesions and involvement of mast cells in the pathological process. PMID- 10813289 TI - The relationship between serum immunoglobulin levels and pulmonary involvement in systemic sclerosis. AB - OBJECTIVE: To examine the relationship between serum immunoglobulin (Ig) levels and pulmonary function in patients with systemic sclerosis (SSc). METHODS: Twenty four patients with SSc who had at least 2 sets of pulmonary function tests (PFT) at intervals of more than one year were eligible. Multiple linear regression models were constructed for prediction of the annualized rates of change of forced vital capacity (FVC), carbon monoxide diffusing capacity (DL(CO)), and DL(CO) per unit alveolar volume (K(CO)). RESULTS: The rates of change of FVC and K(CO) correlated with the annualized rate of change of IgG (p < 0.001 and p = 0.005, respectively), and the rate of change of DL(CO) correlated with the serum IgM level at the first PFT (p = 0.020) and with the annualized rate of change of IgG (p = 0.007). CONCLUSION: The rates of change of serum Ig levels are associated with those of pulmonary function in SSc. Use of this model may assist investigation of pulmonary involvement. PMID- 10813290 TI - Intercellular adhesion molecule 1 gene polymorphisms in polymyalgia rheumatica/giant cell arteritis: association with disease risk and severity. AB - OBJECTIVE: Intercellular adhesion molecule 1 (ICAM-1) is widely distributed in shoulder synovial membrane of active polymyalgia rheumatica (PMR) and strongly expressed in granulomatous inflammatory infiltrate of the temporal artery in giant cell arteritis (GCA). ICAM-1 genes may contribute to the inflammatory PMR/GCA processes. We examined potential associations of ICAM-1 gene polymorphisms with PMR/GCA susceptibility and severity. METHODS: We enrolled 121 consecutive patients with "pure" PMR and 56 patients with biopsy positive GCA residing in Reggio Emilia, Italy. Among patients with PMR, 91 had a followup duration of at least one year. Selected as control subjects were 228 healthy blood donors, 75 patients with nonarteritic central retinal artery occlusion, and 116 cataract surgery patients from the same geographic area. All PMR/GCA patients and controls were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for ICAM-1 polymorphism at codon 241 (exon 4) and codon 469 (exon 6). RESULTS: The frequency of R241 was significantly higher in PMR/GCA patients [p = 0.00001, odds ratio (OR) 5.0 (95% confidence intervals, CI 2.6-9.6) ], in pure PMR patients [p = 0.00001, OR 5.0 (95% CI 2.5-10.0)], and in GCA patients [p = 0.00005; OR 5.0 (95% CI 2.2-11.5)] compared to the healthy controls. The frequency of R241 was significantly higher in total PMR/GCA patients compared to patients with nonarteritic central retinal artery occlusion [p = 0.0007; OR 5.3 (95% CI 1.8-15.5)] and cataract surgery patients [p = 0.0003; OR 4.1 (95% CI 1.8-9.0)]. The distribution of E/K 469 genotype was similar in PMR/GCA patients and in the 3 control groups. Cox proportional hazards modeling identified 2 variables that independently increased the risk of PMR relapse/recurrence: erythrocyte sedimentation rate at diagnosis > 72 mm/h [relative risk 1.6 (95% CI 1.1-2.3)] and the presence of R241 allele [relative risk 1.6 (95% CI 1.1-2.4)]. CONCLUSION: Our findings show that G/R 241 polymorphism of ICAM-1 is associated with PMR/GCA susceptibility and confers an increased risk of relapse/recurrence in PMR. PMID- 10813291 TI - Abnormal blood flow to the submandibular glands of patients with Sjogren's syndrome: Doppler waveform analysis. AB - OBJECTIVE: To assess abnormalities in blood flow to the submandibular glands of patients with Sjogren's syndrome (SS). METHODS: Doppler waveform analysis was performed on the facial artery to assess blood inflow to the submandibular gland of 21 patients with primary SS and 69 healthy subjects. Blood flows were compared before and after secretory stimulation with lemon extracts. RESULTS: The facial artery of healthy subjects in the resting state exhibited a biphasic waveform with a high systolic peak and a prominent second peak of compliance followed by a low diastolic flow. In contrast, the waveform of patients with SS was more uniform compared with the healthy subjects, substantiated by decreased resistive and pulsatility indices, suggesting a hyperemic state of the downstream vascular bed. After stimulation of salivary secretion, the facial artery of healthy subjects responded by decreasing resistive and pulsatility indices, waveform changes indicative of increased blood inflow to the submandibular gland. In contrast, the facial artery of patients with SS responded insufficiently to the stimulation, with the magnitude of changes in the resistive and pulsatility indices being significantly lower than those of the controls. Doppler waveform abnormalities were correlated with the severity of gland damage, supporting a close connection between abnormal blood inflow to the salivary gland and impaired secretory function in SS. CONCLUSION: Our findings suggest that blood inflow responses to secretory stimulation may be defective in the salivary glands of patients with SS. PMID- 10813292 TI - Salivary gland ultrasonography in the evaluation of primary Sjogren's syndrome. Comparison with minor salivary gland biopsy. AB - OBJECTIVE: To evaluate the diagnostic accuracy of ultrasonography (US) of the salivary glands versus histologic examination in detection of salivary gland involvement in primary Sjogren's syndrome (SS). METHODS: Thirty female patients with primary SS diagnosed according to the European criteria and 30 female subjects of similar age with dry mouth (symptomatic controls) due to other selected disorders were studied. Evaluation of salivary involvement was by 2 independent observers for both the parotid and submandibular glands using US score (range 0 to 4) that assigns points to the different degree of glandular inhomogeneity. Lip salivary gland (LSG) biopsies were graded according to the Chisholm and Mason classification system and by morphometric analysis. The area under receiver operating characteristic (ROC) curve was employed to evaluate the screening method's performance. RESULTS: Through ROC curves salivary gland US arose as the best performer, followed by the volume percentage of inflammatory cells and by the semiquantitative focus score. The difference between area under ROC curve of salivary gland US and lymphocytic focus score was significant (p = 0.044), reflecting the accuracy of the imaging diagnostic assessment. No differences arise between areas under the ROC curve of salivary gland US and volume percentage of inflammatory cells (p = 0.610) and between volume percentage of inflammatory cells and semiquantitative focus score (p = 0.081). Attempts to assess the independent contribution of the fractional composition of the LSG biopsy as the predictor of US score showed a significant contribution of both the fractional composition of inflammatory infiltrate (p = 0.01) and intralobular ducts (p = 0.02). These findings indirectly suggest that the US abnormalities we found were strongly related to SS. CONCLUSION: These results conclusively indicate that quantitative assessment of salivary gland ultrasonography is a very useful method fdr evaluating salivary gland involvement in SS and can replace other diagnostic techniques, such as sialography or salivary scintigraphy. PMID- 10813293 TI - Longterm outcome of Wegener's granulomatosis in patients with renal disease requiring dialysis. AB - OBJECTIVE: It is known that renal failure is a poor prognostic marker for survival in Wegener's granulomatosis (WG). We investigated the longterm outcome of patients with WG who have severe renal disease requiring dialysis. METHODS: We performed a retrospective analysis of 104 patients with WG followed at our institution between 1982 and 1997. Twenty-three patients who required dialysis were studied in detail to determine outcomes and factors that influenced survival and restoration of renal function. RESULTS: Of 23 dialysis dependent patients with WG, 11 died (Group 1). 7 either remained dialysis dependent or received successful renal transplants (Group 2), and 5 substantially recovered renal function (Group 3). Mean serum creatinine at the end of a mean followup period of 38.4 months for Group 3 was 1.8 mg/dl. There was no apparent difference between groups in regard to disease profile, e.g., distribution of organ involvement or serum creatinine when renal impairment was first recognized (mean serum creatinine for groups: 1: 3.0 mg/dl; 2: 5.6 mg/dl; 3: 5.5 mg/dl) and peak serum creatinine prior to dialysis (means for groups: 1: 9.5 mg/dl; 2: 10.5 mg/dl; 3: 9.6 mg/dl). Infection secondary to immunosuppression was the leading cause of death in Group I patients. CONCLUSION: Because the clinical profile and degree of renal failure, as judged by serum creatinine, did not differ among patients who did or did not regain dialysis independent renal function, we recommend aggressive immunosuppressive therapy in all cases of active WG with acute rapidly worsening renal failure, regardless of the severity of renal impairment. PMID- 10813294 TI - Microscopic inflammatory changes in colon of patients with both active psoriasis and psoriatic arthritis without bowel symptoms. AB - OBJECTIVE: To evaluate colonic mucosa of patients with both active psoriasis and psoriatic arthritis (PsA) without bowel symptoms. METHODS: Fifteen persons (9 men, 6 women) who had both active psoriasis and PsA without bowel symptoms underwent colonoscopy with multiple biopsies of bowel mucosa. Ten nonhospitalized healthy subjects in followup colonoscopy after resection of benign polyps (8 men, 2 women) took part as a control group. RESULTS: Six psoriatic patients (40%) showed macroscopically normal colonic mucosa. In the remaining 9 reddening was frequently recorded (6 cases). while edema and granular changes appeared less commonly (3 cases each, respectively). Friability was markedly rare (only one case) and bleeding and ulcerations were absent. All 15 patients showed microscopic changes. Increase in lamina propria cellularity (consisting of plasma cells and lymphocytes) and lymphoid aggregates were found in all cases. Active inflammation, evident as neutrophilic polymorph infiltration occurred in 9 patients. Glandular atrophy was found in 3 cases; mucosal surface changes and crypt abnormalities occurred in one case each. No control had macroscopic or microscopic inflammatory changes of bowel mucosa. CONCLUSION: Bowel mucosa of patients with PsA without bowel symptoms show microscopic lesions even when mucosa appeared macroscopically normal. This result may support a pathogenetic link between skin, joints, and gut in psoriatic patients with arthritis even in the absence of bowel symptoms. PMID- 10813295 TI - The epidemiology of psoriatic arthritis in Olmsted County, Minnesota, USA, 1982 1991. AB - OBJECTIVE: To determine the incidence, prevalence, and outcomes of psoriatic arthritis (PsA) in a geographically defined community. METHODS: Using the Rochester Epidemiology Project computerized medical record system, we screened all records of Olmsted County, Minnesota, residents with any diagnosis consistent with psoriasis and/or PsA made between January 1, 1982 and December 31, 1991. Medical records were reviewed using a pretested data collection form. Only those cases of psoriasis where the diagnosis was confirmed by a dermatologist were included. PsA was defined as inflammatory arthritis associated with a definite diagnosis of psoriasis. All identified cases were followed until death, migration from the county, or January 1, 1992. Cases with seropositive rheumatoid arthritis, systemic lupus erythematosus, crystal induced arthritis, Reiter's syndrome, arthritis associated with inflammatory bowel diseases, and inflammatory osteoarthritis were excluded. Clinical characteristics were described using summary statistics. Age and sex adjusted incidence and prevalence rates were calculated. Survival was estimated using the Kaplan-Meier method. RESULTS: We reviewed the records of 1844 patients with a diagnosis of psoriasis. In 1056 of these, the diagnosis was confirmed by a dermatologist. Among these 1056 psoriasis cases, we identified 66 cases (34 female, 32 male) of PsA first diagnosed between 1982 and 1991. The average age and sex adjusted incidence rate per 100,000 US population was 6.59 (95% confidence interval, CI, 4.99, 8.19) and the prevalence on January 1, 1992, was about one per 1000 (95% CI 0.81, 1.21). The average age at diagnosis was 40.7 years. At diagnosis, 91, 3, and 6% of cases had oligoarthritis, polyarthritis, and spondylitis, respectively. Over the 477.8 person-years of followup, 25 developed extraarticular manifestations (enthesitis, n = 15; ocular inflammation, n = 11; urethritis, n = 9), 10 patients received disease modifying antirheumatic drug treatment (methotrexate, n = 7; sulfasalazine, n = 5; intramuscular gold, n = 1; oral gold, n = 1), 3 received corticosteroids, and 5 had surgical interventions (synovectomy, n = 3; arthroplasty, n = 1; other reconstructive surgery, n = 2). Survival was not significantly different from the general population (p = 0.546). CONCLUSION: Unlike results from previous referral based studies, our findings indicate that PsA is a mild, uncommon inflammatory arthritis, not associated with a significant increase in mortality. PMID- 10813296 TI - Sensitivity to change of mobility tests; effect of short term intensive physiotherapy and exercise on spinal, hip, and shoulder measurements in spondyloarthropathy. AB - OBJECTIVE: To assess the sensitivity to change of 13 spinal, shoulder, and hip measurements and determine correlation between age, disease duration, and radiologic findings in ankylosing spondylitis (AS) and other spondyloarthropathies (SpA). METHODS: We assessed 112 patients with various types of SpA as changes before and after an intensive inpatient course. RESULTS: The most sensitive spinal measurements were finger to floor distance, chest expansion. thoracolumbar rotation (TLR), and lateral flexion, in that order. Cervical rotation, lateral flexion, and intermalleolar distance might also be useful in short term clinical trials, while the Schober tests, thoracolumbar flexion, and occiput-wall distance were not sufficiently sensitive. Hip internal rotation (HRi), shoulder flexion, and abduction measurements were also sensitive, though possibly more suitable for patients with articular symptoms. TLR and HRi were the only measurements that correlated markedly with disease duration, but not with age. Changes in the measurements correlated significantly with radiological spinal changes. CONCLUSION: Finger to floor distance, TLR, and thoracolumbar lateral flexion were the most sensitive to detect improvements in short term clinical trials, while the Schober test, thoracolumbar flexion, and occiput-wall distance were insensitive. PMID- 10813298 TI - Isometric progressive resistive exercise for osteoporosis. AB - OBJECTIVE: To assess the effect of site-specific resistive isometric exercises on muscle strengthening of 10 muscle groups over 2 months. A second study measured bone alkaline phosphatase (ALP) as a marker of bone formation and bone resorption [urine deoxypyridinoline (D-Pyr) crosslinks] in a similar cohort. METHODS: Twenty postmenopausal Caucasian women aged 56 to 69 yrs (mean 61) on a stable or no medication regimen for the previous 6 months participated in the initial exercise cohort. Twenty-one women ages 52-69 (mean 62) participated in the second cohort. All women had osteopenia (DXA T less than -1.0) or osteoporosis (DXA T less than 2.5). An inflatable ball with attached nonelastic straps provided progressive resistance. Exercises consisted of a 5 s maximum contraction against progressively increasing resistance of the ball or nonelastic straps. Muscle strengthening was measured by a hand held dynamometer at 0, 4, and 8 weeks. Twenty women completed 8 weeks of the initial study and 21 women completed the second study. RESULTS: Muscles showing increased strength in the first cohort were neck extensors (p < 0.04), hand grips (p < 0.02), elbow flexors (p < 0.05), quadriceps (p < 0.04), trunk extensors (p > 0.05, not significant = NS). Elbow extensors were purposely not exercised (as a control) and showed no significant strength increase. In the second cohort, increased muscle strength was measured in the neck extensors (p < 0.001), trunk extensors (p < 0.001), and left quadriceps (p < 0.024); and bone ALP increased (p < 0.05), with no change detected in bone resorption (urine D-Pyr). CONCLUSION: Brief progressively resisted isometric exercises for 10 min daily are an adequate stimulus for muscle strengthening of the neck, back, upper and lower extremities, and are capable of enhancing bone formation measured by bone ALP. PMID- 10813297 TI - HLA-B27 has no effect on the phenotypic expression of progressive ankylosis in ank/ank mice. AB - OBJECTIVE: To investigate whether HLA-B27 influences the expression of murine progressive ankylosis (MPA), a single-gene autosomal recessive mouse model of ankylosing spondylitis that arises in mice homozygous for the ank gene. METHODS: Mice transgenic for HLA-B27 were bred with ank/ank mice, and the phenotypes of the F1 and F2 progeny were observed. RESULTS: ank/+ mice showed no abnormalities, and ank/ank mice showed the typical phenotype of MPA, irrespective of B27 status. CONCLUSION: HLA-B27 and the ank/ank genotype both predispose to diseases involving progressive bony ankylosis. These findings suggest that these disease processes are distinct and noninteractive, and they provide no support for the hypothesis that the human homolog of the ank locus participates in the pathogenesis of ankylosing spondylitis. PMID- 10813300 TI - Factors of importance for work disability in women with fibromyalgia. AB - OBJECTIVE: To identify factors of importance for women with fibromyalgia (FM) to continue working despite the limitations imposed by the symptoms. METHODS: A mail questionnaire with questions regarding social background, symptoms, sickness benefits, work situation, work conditions and adjustments, opinions regarding own work ability, and satisfaction with the situation was sent to 218 consecutive women seen at a university pain or rheumatology clinic. Answers were obtained from 176 women. RESULTS: Pain, poor quality sleep, abnormal tiredness, muscle stiffness, and increased pain after muscle exertion were frequently reported symptoms. Fifty percent of the women were employed, 15% full-time. Twenty-three percent reported FM as the reason for not working. The work situation had been changed for 58% of the working women, and 80% counted on being able to continue working. CONCLUSION: Work disability is a serious concern in FM, and the majority of women with FM have limitations in their ability to work. Our results indicate that individual adjustments in the work situation need to be made and that women who have found a level matching their ability may continue to work and find it satisfactory. Early intervention in the work situation is recommended. PMID- 10813299 TI - Glucocorticoid dose dependent downregulation of glucocorticoid receptors in patients with rheumatic diseases. AB - OBJECTIVE: The therapeutic success of low doses of glucocorticoids is mediated entirely by classical genomic effects, whereas that of high doses is also mediated to an as yet unknown extent by nongenomic effects. We assessed the relative therapeutic importance of these nongenomic effects in pulse therapy. METHODS: A [3H]dexamethasone radioligand binding assay was used to measure the number of glucocorticoid receptor sites (R, given as number of sites per cell) and glucocorticoid receptor binding affinity (Kd, given in nM) in peripheral blood mononuclear cells isolated from 26 healthy control blood donors and 27 patients with rheumatic diseases. Patients were divided into 4 groups on the basis of their glucocorticoid dose: 0 mg (Group A), < or = 0.25 mg (Group B), 0.25 to 1 mg (Group C), and > 1 mg (Group D) of prednisolone equivalent per kg per day. RESULTS: Sex independent normal values of 3605 +/- 1136 for R and 5.39 +/- 3.4 for Kd were found. At 5407 +/- 1968, the number of receptor sites in patients not receiving glucocorticoid therapy (Group A) was significantly higher than that of controls (p < 0.01). In patients receiving glucocorticoid therapy this value was reduced at 3855 +/- 866 (Group B), 3358 +/- 963 (Group C), and 2685 +/- 962 (Group D). The values in Groups C and D were significantly lower than those in untreated patients (p < 0.02). CONCLUSION: In pulse therapy doses of glucocorticoids that exceed receptor saturation are administered for several days, but in addition significant receptor downregulation occurs. Therefore, we assume an increase in the relative contribution of the nongenomic effects of glucocorticoids to the therapeutic success under these conditions. PMID- 10813301 TI - Force production capacity and acute neuromuscular responses to fatiguing loading in women with fibromyalgia are not different from those of healthy women. AB - OBJECTIVE: To compare the maximal and explosive strength characteristics of the leg muscles in premenopausal women with fibromyalgia (FM) with those of healthy female controls (HC) and to examine acute neuromuscular fatigue during heavy resistance loading and short term recovery from fatigue in these 2 groups. METHODS: Subjects were 11 women with FM, 38.6 (5.8) years old, and 12 healthy female controls, 37.3 (6.1) years old. The following were recorded before, during, and after a fatiguing loading session: maximal bilateral concentric and isometric force, isometric force-time curves and relaxation-time curves with agonist-antagonist neural activation (by EMG) of the leg muscles, muscle pain, and blood lactate concentrations. RESULTS: At baseline all the measured muscle strength characteristics were comparable between the study groups. The heavy fatiguing loading led to considerable and comparable acute fatigue found in both muscle strength characteristics and agonist-antagonist electromyography in both groups. The respective changes in blood lactate concentration and subjectively perceived muscular pain in the loaded muscles during strenuous resistance loading and recovery from fatigue were similar in both groups. CONCLUSION: Premenopausal women with FM do not demonstrate lower dynamic or isometric muscle strength characteristics compared to matched healthy controls. Second, the similar neuromuscular responses recorded during and after the fatiguing loading strongly support the hypothesis of normal muscle structure and neuromuscular function in patients with FM. PMID- 10813302 TI - Comparison of criteria for the classification of childhood arthritis. AB - OBJECTIVE: To evaluate the applicability of the ILAR criteria for classification of childhood arthritis in an outpatient pediatric rheumatology clinic population, and to determine the proportion of children who met standard classification criteria, but failed to meet ILAR criteria for specific arthritides, and therefore became unclassifiable. METHODS: We reviewed the charts of 70 consecutive patients who had arthritis for at least 6 months, and attended the clinic between September and November 1997. Sixty-nine patients were categorized according to one of the traditional classifications [ACR for juvenile rheumatoid arthritis (JRA), European Spondylarthropathy Study Group (ESSG) for spondyloarthropathy, Vancouver Criteria for juvenile psoriatic arthritis (JPsA)], and the ILAR classification system. RESULTS: Sixty-one patients (88.4%) were classifiable by the ILAR system; 8 others failed to fulfill ILAR criteria for any specific category, and were assigned to the "other arthritis" category. Of the 29 patients with oligoarticular onset JRA, 6 were unclassified, 5 because of exclusions, and one because he fulfilled criteria for 2 categories. Presence of a family history of psoriasis accounted for most of the exclusions in the oligoarthritis and enthesitis related arthritis categories. All patients with polyarticular onset or systemic onset JRA were classified in the corresponding category in the ILAR system. One 9-year-old patient with spondyloarthropathy was reclassified as "other arthritis" because of exclusions. All 6 children with definite JPsA met ILAR criteria for PsA. Of 4 patients with probable JPsA, only 2 met ILAR criteria for PsA, a third was classified as rheumatoid factor negative polyarthritis, and the fourth was classified as "other arthritis" because of exclusions. CONCLUSION: The ILAR classification criteria applied to a group of children with chronic arthritis classified by traditional criteria results in reassignment of 11.6% of the patients, predominantly in the oligoarticular group. It will be important to determine the role of the presence of a family history of psoriasis in classifying these patients. PMID- 10813303 TI - Effects on bone metabolism of one year recombinant human growth hormone administration to children with juvenile chronic arthritis undergoing chronic steroid therapy. AB - OBJECTIVE: To study the effects on bone metabolism of treatment with recombinant human growth hormone (rhGH) in children with juvenile chronic arthritis (JCA) who are undergoing treatment with glucocorticoids (GC) and have severe bone lesions. METHODS: We assessed the effects of rhGH treatment (1.4 U/kg/week) on bone metabolism markers and bone density measured during a one year treatment course in 14 patients with systemic forms of JCA undergoing longterm GC treatment. RESULT: All patients at inclusion showed severe bone demineralization (mean bone density: -3.7 standard deviation score for chronological age). Compared to pretreatment values, bone formation markers (blood levels of osteocalcin and C terminal propeptide of type 1 procollagen) and bone resorption markers (urinary hydroxyproline, pyridinoline, and deoxypyridinoline levels) increased significantly during treatment and returned to pretreatment values after discontinuation of rhGH. We observed that plasma level of osteocalcin was the best predictive variable of growth response to rhGH treatment in these patients. CONCLUSION: The results reflect an increase in bone turnover in these patients. Despite these biochemical changes no improvement of bone density was observed during the one year treatment. Treatment of longer duration is necessary to evaluate the curative effects of GH. PMID- 10813304 TI - X-ray absorptiometry of bone in obese and eutrophic children from Valparaiso, Chile. AB - OBJECTIVE: To assess and compare bone mineral content (BMC) and bone mineral density (BMD) by dual X-ray absorptiometry (DEXA) measurement in obese and eutrophic children. METHODS: In a cross sectional, case control study 16 obese children (8 male, 8 female) aged 5 to 13 years were recruited from the outpatient nutrition clinic of Carlos van Buren Hospital, Valparaiso, Chile, during 1997. Sixteen healthy eutrophic children matched for sex, chronological age, height, and pubertal stage were enrolled as controls. The mean dietary calcium intake was 580 +/- 100 mg/day. All obese patients had more than 2 standard deviation (SD) of height/weight ratio. Lumbar spine (L2-L4) BMD, femoral neck BMD, total body BMD, and total body BMC were measured by posteroanterior dual photon DEXA. The results were expressed as mean +/- SD. Comparison of the data was determined by Wilcoxon and Student t test. RESULTS: Mean BMC was 1684.1 +/- 492.38 g in obese children and 1418.2 +/- 483.48 g in controls (p < 0.001, Student t test; p = 0.001, Wilcoxon test). Mean total BMD was 817.5 +/- 99.37 g/cm2 in obese children and 771.62 +/- 105.62 g/cm2 in controls. (p = 0.041 by Wilcoxon). Mean hip BMD was 784.4 +/- 117.05 g/cm2 in obese children and 801.67 +/- 150.34 g/cm2 in controls. Mean spine BMD was 724.87 +/- 171.75 g/cm2 in obese children and 686 +/- 137.08 g/cm2 in controls (not significant). CONCLUSION: Obese children have more total body BMC than eutrophic children. There was no significant difference in regional hip BMD and lumbar spine BMD among obese and normal children. Obese children may have larger bones. PMID- 10813305 TI - Primary angiitis of the central nervous system presenting as a mass lesion in a child. PMID- 10813306 TI - Carpal tunnel syndrome caused by Mycobacterium szulgai. AB - We describe 2 cases of infection due to Mycobacterium szulgai revealed by a carpal tunnel syndrome (CTS) that was the only clinical manifestation. Both patients regularly cleaned their fish tank with bare hands. The diagnosis was made by isolation of M. szulgai from synovium. The cause of the CTS was a synovitis. Previous synovectomies were ineffective. Improvement was observed with antibiotic treatment. The only way to diagnose this unusual infection is to perform histology of synovium and to isolate the mycobacteria from synovium culture. PMID- 10813307 TI - Severe xerostomia in a young man. AB - We describe a previously healthy 24-year-old male waiter who presented with very severe xerostomia, such that he consumed an average of 7 pints of milk a day and seriously considered suicide. There were no other symptoms or signs of exocrine gland dysfunction. We discuss the differential diagnosis, investigation, and his subsequent complete response to therapy, with a review of the literature. PMID- 10813308 TI - Nocardiosis in patients with systemic lupus erythematosus. The Singapore Lupus Study Group. AB - Nocardia, a gram positive variably acid-fast aerobic bacterium is an opportunistic pathogen in immunocompromised hosts. We present 5 cases of nocardiosis in patients with systemic lupus erythematosus. We emphasize the clinical features, radiologic findings, and antibiotic sensitivity. Lung involvement was the predominant manifestation; others include brain abscess, retinitis, thyroiditis, and diaphragmatic infiltration. We describe the first cases of pulmonary nocardiosis presenting as pneumothorax and the use of fine needle aspiration cytology in diagnosing nocardial thyroiditis. PMID- 10813309 TI - Femoral neuropathy secondary to ossification of the ligamentum flavum. AB - Radiculopathy resulting from ossification of the ligamentum flavum (OLF) is extremely rare and concerns only intercostal neuralgias. We describe a 37-year old Caucasian woman with a lumbar radiculopathy revealing an OLF. Her symptoms were completely and definitively relieved by surgery. PMID- 10813310 TI - Chronic pain in a family of 6 in the context of litigation. AB - We describe a family of 6 (2 parents and 4 children), evaluated 6 and 8 years after a minor car accident for chronic pain. A near identical complex of multiple physical, constitutional, and psychological symptoms were shared by all family members, all of whom bore the diagnosis of fibromyalgia. The case was brought to court after nearly a decade of symptomatology and extensive use of the health care system. The minor compensation awarded was consumed ultimately by legal fees. Psychosocial/personality issues and iatrogenic and medico-legal contributions in the evolution and resolution of the legal claim are discussed. PMID- 10813311 TI - Chronic recurrent multifocal osteomyelitis showing marked improvement with corticosteroid treatment. PMID- 10813312 TI - Dramatic aneurysm regression in polyarteritis nodosa following high dose pulse cyclophosphamide. PMID- 10813313 TI - Complications of NSAID gastropathy and use of gastric cytoprotection: experience at a tertiary care health center. PMID- 10813314 TI - Human immunodeficiency virus infection and arthritis. PMID- 10813315 TI - Premature coronary artery disease in systemic lupus erythematosus in the absence of corticosteroid use. PMID- 10813316 TI - Sjogren's syndrome and multiple amyloidomas. PMID- 10813317 TI - Henoch-Schonlein purpura in adults and autoimmune thyroiditis. PMID- 10813318 TI - Differential sugar absorption as a marker for adaptation in short bowel syndrome. AB - PURPOSE: There are no reliable monitoring methods for following up with patients with short bowel syndrome (SBS). This study examines the use of inert sugar markers (mannitol and lactulose) as indicators of the surface area increases occurring with adaptation. METHODS: Juvenile male rats underwent either transection with intestinal reanastomosis or resection with removal of the proximal 90% of the small bowel, leaving 10 cm of terminal ileum (n = 8 in each group). Animals were studied in vivo, measuring absorption of mannitol and lactulose on day 7, 14, and 28 and killed with the representative histological samples taken on day 7, 14, and 28. RESULTS: Resected animals showed significant increases in intestinal length (initial bowel length 10 cm, final length 17.8 +/- 1.4 cm, while transected showed no significant changes (101 +/- 2 cm): resection also increased intestinal circumference (initial circumference 0.5 cm +/- 0.1 cm, final circumference 1.1 cm +/- 0.2 cm in resected animals, while transected animals remained unchanged). Resected animals also showed significant increases in villus height (0.7 +/- 0.06 mm initially, 0.9 +/- 0.09 mm at day 28), and but a decrease in villus density (116 +/- 20 villi/mm2 initially, 78 +/- 20 villi/mm2 at day 28), again controls showed no changes. This resulted in a significant increase in intestinal surface area in resected animals over the study; surface area initially calculated at 640 +/- 80 cm2 increasing to 1,440 +/- 360 cm2, while controls showed no significant change in surface area. There was also an increase in mannitol absorption, which went from 1.8 +/- 0.6% on day 7 to 2.56 +/ 0.6% on day 28 in resected animals, while permeability actually decreased over time. (All data mean +/- SD, with P<.05 by Student's t test). Mannitol absorption correlated well with intestinal surface area (R2 = 0.82). CONCLUSION: These results suggest that inert sugar markers, such as lactulose and mannitol, may be useful in following adaptation in patients who have short bowel syndrome. PMID- 10813319 TI - Near-total intestinal aganglionosis: long-term follow-up of a morbid condition. AB - PURPOSE: The aim of this study was to understand the long-term outcome of near total intestinal aganglionosis (NTIA). METHODS: The study was an institutional review of all patients with NTIA between 1974 and 1999, inc!usive. Seven full term babies were identified (4 boys, 3 girls). Mean birth weight was 3.2 kg (range, 2.7 to 4.1). Patients included 1 pair of siblings and 1 Down's syndrome baby. All 7 had obstruction within 5 days of life. One patient experienced bowel perforation. All 7 had NTIA diagnosed within the first 2 months of life. In 2 of 7, aganglionosis began 10 cm distal to the ligament of Treitz (LOT). The longest segment of normal small bowel was 130 cm from the LOT. Five of 7 babies underwent a stoma between day of life (DOL) 1 and 15. RESULTS: Two of three "long-term" survivors (>2 years) had corrective surgery by 13 months. The first received extended jejuno-ileal myotomy and myectomy with multiple postoperative problems. She died at 8 years of age of complications of total parenteral nutrition (TPN). The second underwent myotomy, resection and patch graft to jejunum, and several jejunostomy revisions. All of the involved bowel eventually was resected. The 3 of 7 who lived more than 1 year all took some nutrition orally. Three died, between 3 months and 8 years of age, of complications of TPN. There are 2 survivors at 3 and 7 years of age. Survival has not correlated with the length of aganglionic bowel. CONCLUSIONS: Aganglionosis involving most of the bowel has a high morbidity and mortality rate. Since 1990 a more aggressive surgical approach has resulted in improved survival rates but with significant morbidity. For children surviving beyond 3 months of age, outcome was less dismal. Some patients may benefit from extended jejunal myotomy or myectomy. However, postoperative complications are the rule, not the exception. PMID- 10813320 TI - Idiopathic gastric perforation in neonates and abnormal distribution of intestinal pacemaker cells. AB - BACKGROUND/PURPOSE: The etiology of idiopathic gastric perforation (IGP) in neonates is unclear. Interstitial cells of Cajal (ICC) express tyrosine kinase receptor C-kit, and act as gastrointestinal pacemaker cells. Stem cell factor (SCF) is a C-kit ligand and plays an important role in immune system homeostasis in the gastrointestinal tract. The authors hypothesized that abnormal distribution of ICC or SCF in the gastric wall (ie, abnormal motility or impaired immunity) could predispose the stomach to IGP. METHODS: Stomachs obtained at postmortem from neonates who died of IGP (n = 7) and other causes (control group; n = 10) were used. Biopsy sections were taken at random from various sites in the stomach, including macroscopically intact areas, and labeled immunohistochemically using antibodies to C-kit(a marker for ICC) and SCF. RESULTS: In all control specimens, ICC were present between the muscle layers and around the myenteric plexuses of the stomach wall. In contrast, ICC were absent in all biopsy sections from 3 of the 7 IGP stomachs. In the remaining 4 IGP stomachs, there were fewer ICC in the muscle layers compared with controls, and ICC were absent around the myenteric plexuses. The distribution of SCF immunoreactivity in IGP and control specimens was similar. CONCLUSION: The findings suggest that a lack of ICC (ie, gastric hypomotility) may be implicated in the etiology of IGP in neonates. PMID- 10813321 TI - Local staging of Wilms' tumor--computerized tomography correlation with histological findings. AB - PURPOSE: The aim of this study was to assess the accuracy of computed tomography (CT) in the local staging of Wilms' tumor. METHODS: Twenty-six patients with unilateral, nonmetastatic Wilms' tumors were accrued over the period of 1990 through 1997. Preoperative CT staging was performed before surgical biopsy or excision. The abdominal CT images were reviewed and staged for local disease according to National Wilms' Tumour Study (NWTS) Group Staging V as either intrarenal disease (Stage I), local extrarenal extension amenable to complete local excision (Stage II), or advanced local disease (Stage III). Patients with metastatic (Stage IV) and bilateral disease (Stage V) were excluded from our study. Results were compared to histological findings. RESULTS: CT scan correctly staged 10 of 26 cases (38%) of Wilms' tumor. CT scan overstaged 12 of 16 (75%) localized renal disease (Stage I) and local extension (Stage II) tumors, and understaged 4 of 10 (40%) localized spread (Stage III) tumors. CONCLUSIONS: CT scan appears to have poor correlation to histological staging. Therefore, basing therapy solely on radiological imaging may lead to some patients receiving more intense therapy than necessary, whereas others do not receive sufficient therapeutic regimens. Histological assessment of the excised mass should remain the standard for staging Wilms' tumors. PMID- 10813322 TI - Complications of laparoscopic treatment of esophageal achalasia in children. AB - BACKGROUND/PURPOSE: The aim of this study was to evaluate the incidence and management of the complications that occurred in some children who underwent laparoscopic Heller's esophagocardiomyotomy in the authors' institutions. METHODS: Between March 1993 and October 1998, the files of all the children with achalasia who underwent laparoscopic Heller's esophagocardiomyotomy in a community hospital in Naples, Italy, and a private hospital in Paris, France, were reviewed. A 5-port technique was used associating Heller's esophagocardiomyotomy to an antireflux surgical mechanism (Dor's or Toupet's) in all cases. Intra- and postoperative complications, as well as the postoperative outcome, were evaluated. RESULTS: Ten laparoscopic Heller's esophagocardiomyotomies were performed in 5 girls and 5 boys with achalasia. Age ranged between 2 and 13 years. Mean operating time was 120 minutes. Hospital stay ranged between 3 and 41 days. Complications were recorded in 3 patients: in 2 an esophageal mucosal perforation and in 1 a prolonged dysphagia. Two of these complications occurred in the last patients operated on. Follow-up varied from 6 months to 6 years. All children were free of symptoms. CONCLUSIONS: The results show that laparoscopic Heller's esophagocardiomyotomy in children is a feasible procedure. Assessment of mucosal integrity immediately after the myotomy must be performed. Complications can happen even if the operation is performed by expert laparoscopic surgeons. PMID- 10813323 TI - Long-term disability after trauma in children. AB - PURPOSE: The aim of this study was to assess long-term disability after pediatric trauma and identify predicting factors. METHODS: A phone survey was conducted of all pediatric trauma patients with an Injury Severity Score (ISS) > or = 4 seen over 6 years at a regional trauma center. The questionnaire was a modification of the Rand Health Insurance Study scales. RESULTS: Of 218 eligible trauma registry patients, 19 were deceased, 64 unreachable, 19 declined, and 116 interviewed. There were no demographic differences between respondents and nonrespondents. Sixty-three percent of the respondents were boys, mean age at injury was 13, ISS 16.7, and mean interval since injury was 4.4 (range, 1 to 7) years. Sixty-three children (54%) had no limitations on follow-up; the remainder had either limitations in physical or role activities (28%), mobility (16%), or self-care ability (2%). Significant correlations were found between the presence of disability and trauma scores and number of body regions injured. Stepwise logistic regression identified number of regions injured, mechanism of injury, and ISS as the main determinants for presence of long-term disability. CONCLUSIONS: Half of injured children do have long-term sequelae. Their occurrence can be predicted from trauma scores, mechanism of injury, and number of regions injured. PMID- 10813324 TI - Trends in bicycling-related head injuries in children after implementation of a community-based bike helmet campaign. AB - BACKGROUND/PURPOSE: The aim of this study was to determine the effect of a community-based bike helmet promotion campaign on bike helmet use and related head injuries in children (0 to 14 years of age) in a large North American city. METHODS: The authors established a multifaceted, multidisciplined, community based campaign to promote bike helmet use by children in 1989. The goals were to increase helmet use by 100% per year, to reduce fatal bike-related head injuries by 50% overall, and to explore the feasibility of legislation mandating helmet use. Helmet use was measured by standardized field observations repeated annually in a single borough within the metropolitan area. To estimate head injury incidence, the number of admissions to hospital for the treatment of bike-related head injuries in a regional trauma registry, which included all residents in the target population was used. The authors were unable to control for changes in exposure to bicycling or in the criteria for admissions to hospital for the treatment of head injuries during the study period. RESULTS: The bike helmet use rate rose from 4% in 1990 to 67% in 1996. The number of head injury admissions fell from 46 in 1990 to 24 in 1996. Legislation requiring helmet use by all children went into effect in October 1995. CONCLUSIONS: Bike helmet use increased significantly during the first 4 years of the campaign and again after the helmet law was implemented. The total number of bike-related head injury admissions declined by more than 50%. The campaign achieved all of its goals except for a 50% reduction in fatal head injuries, which were too infrequent for analysis. PMID- 10813325 TI - The late nonfunctioning duodenal atresia repair--a second look. AB - BACKGROUND/PURPOSE: In 1986, the authors reported on 3 newborns who had repair of their duodenal atresia, and between 6 and 18 months postoperatively an anastomotic obstruction developed in each suddenly. After prolonged medical and surgical treatments it became apparent that the duodenal atresia repair was functionally obstructed and plication of the dilated atonic proximal duodenum was curative. Since then, 2 more patients became so obstructed at 5 and 24 years postoperatively. The aim of this study was to report the very late occurrence of a functional obstruction of a newborn duodenal atresia repair. METHODS: The 2 additional histories and surgical repairs were reviewed. RESULTS: The 5-year old boy was cured immediately with plication only of his dilated proximal duodenum. The 24-year-old nurse had a very stormy 2-year course with several bypass operations, which did not relieve her abdominal pain and bile vomiting until they were taken down and her dilated proximal duodenum was plicated, after which she made a good recovery. Both remain well. CONCLUSION: An uncommon, very late, sudden, apparently anastomotic, postoperative, newborn, duodenal atresia repair obstruction caused by proximal, dilated, duodenal atony, and dysfunction can occur many years later and responds to duodenal plication alone. PMID- 10813326 TI - Effect of growth hormone, epidermal growth factor, and insulin on bacterial translocation in experimental short bowel syndrome. AB - BACKGROUND/PURPOSE: An adaptive process starts in the remaining intestine after massive resection, and several trophic factors including growth hormone (GH), epidermal growth factor (EGF), and insulin (INS) have been shown to have a positive effect on it. Bacterial translocation (BT) is frequent after extensive small bowel resection, but the effects of GH, EGF, or INS have not been investigated in experimental short bowel syndrome (SBS). This study tests the hypothesis that GH, EGF, or INS decrease BT in SBS in rats with parenteral nutrition (PN). METHODS: Thirty-eight adult Wistar rats underwent central venous cannulation and were assigned randomly to 1 of 4 groups receiving for 10 days 4 treatment regimes: (1) PN group (n = 10): fasting, all-in-one PN solution (300 mL/kg/24 h, 280 kcal/kg/24 h), 80% gut resection including ileo-cecal valve; (2) GH group (n = 9): fasting, same PN regime and resection, GH (1 mg/kg/d, subcutaneously); (3) EGF group (n = 9): fasting, PN, resection, EGF (150 microg/24 h intravenously); (4) INS group (n = 9): fasting, PN, resection, INS (1 UI/100 g/24 h subcutaneously). At the end of the experiment they were killed, and mesenteric lymph nodes (MLN) and peripheral and portal blood samples were recovered and cultured. Several fragments of intestine were taken to determine cell proliferation (PCNA index) and morphometric parameters (villous height, crypt depth). RESULTS: GH, EGF, and INS groups showed a 28%, 29%, and 30% increase in gut mucosal thickness, and PCNA index rose 21%, 20%, and 25%, respectively in comparison with PN controls. Bacterial translocation to peripheral blood was detected in 0% of PN animals and in 44%, 40%, and 28% of GH, EGF, or INS rats, respectively (P < .05). No differences were found in BT in MLN or portal blood among groups. CONCLUSION: Administration of GH, EGF, or INS improves gut mucosal structure in rats with SBS under PN, but, surprisingly, the incidence of BT detected in peripheral blood was increased rather than decreased in animals receiving these treatments. PMID- 10813327 TI - Repair of the huge umbilical hernia in black children. AB - Surgical repair of large umbilical hernias may present a challenging surgical problem. The currently described surgical techniques often yield disappointing results. The authors describe a new technique that allows for the repair of the fascial defect and the creation, with the use of a square cutaneous flap, of a neoumbilicus with sufficient depth and a good cosmetic appearance. PMID- 10813328 TI - Laparoscopic versus open splenectomy in children. AB - BACKGROUND: The authors have reviewed their initial experience with laparoscopic splenectomy (LS) to identify the indications, success rate, and complications associated with this procedure compared with a series of children undergoing open splenectomy (OS) during the same time period. METHODS: The records of 51 children who underwent splenectomy from 1993 through 1998 were reviewed retrospectively. RESULTS: Thirty-five patients aged 1 to 17 years (mean, 9.4 years) underwent LS for the following indications: ITP (n = 20), sickle cell disease or thalassemia (n = 6), hereditary spherocytosis (n = 5), other hematologic disorders (n = 4). Seventeen patients aged 2 to 17 years (mean, 11.8 years) underwent OS during the same time period for ITP (n = 4), sickle cell disease or thalassemia (n = 4), hereditary spherocytosis (n = 5), and other indications (n = 4). Concomitant cholecystectomy was performed in 4 of 35 LS and 4 of 17 OS. Accessory spleens were identified in 10 of 35 LS and 2 of 17 OS cases. Eleven spleens were enlarged in the LS group, and 8 were enlarged in the OS group. One LS required conversion to an open procedure because the spleen did not fit in the bag. No other cases were converted. Median estimated blood loss was 50 mL for both the LS and OS groups. The only intraoperative complication in the LS group was a splenic capsular tear, which had no effect on the successful laparoscopic removal of the spleen. No patient in either group required a blood transfusion. The LS patients had a shorter length of hospital stay (1.8 +/- 1 versus 4.0 +/- 1 day, P = .0001). Total hospital charges were not significantly different. Follow-up ranged from 6 to 40 months. One LS patient died 47 days postoperatively from unrelated causes. Two LS patients had recurrent ITP; accessory spleens were found in one and resected laparoscopically. CONCLUSION: LS in children can be performed safely with a low conversion rate (2.9%) and is associated with a shorter hospital stay and comparable total hospital cost when compared with OS. PMID- 10813329 TI - Pediatric perineal impalement injuries. AB - Pediatric perineal impalement injuries are relatively uncommon. There may be difficulty in recognizing or properly treating such injuries, because their severity may not be reflected accurately by the external appearance of the perineum. The authors describe 3 case reports of patients with perineal impalement injuries and their management. The authors emphasize how a thorough workup can prevent missed injuries, leading to timely surgical repair and good outcomes. PMID- 10813330 TI - Emergent abdominal decompression with patch abdominoplasty in the pediatric patient. AB - BACKGROUND/PURPOSE: Abdominal compartment syndrome (ACS) is the cardiac, pulmonary, and renal dysfunction that occurs as a result of elevated intraabdominal pressure. The authors present their experience with patch abdominoplasty (PA) in pediatric patients as a means to treat and prevent ACS. METHODS: The charts of patients who underwent PA were reviewed retrospectively. ACS was defined as the increased oxygen requirements and elevation of peak inspiratory pressures (PIP) associated with abdominal distension and worsening renal and or cardiac function. RESULTS: A total of 23 patients (13 boys) were treated (average age, 23 months). Diagnoses included necrotizing enterocolitis (NEC, n = 13), trauma (n = 3), Hirschsprung's enterocolitis (n = 2), perforated bowel (n = 4), and bilateral Wilms' tumor with bowel obstruction (n = 1). Oxygen requirements decreased after patch abdominoplasty (mean preoperative FIO2, 0.87 +/- 24, mean postoperative, 0.67 +/- 24 [P = .01]). The PIP decreased significantly in the 13 patients who survived (mean preoperative PIP, 33 +/- 8, mean postoperative PIP, 27 +/- 7 [P = .01]). These PIPs failed to respond in the 8 nonsurvivors (mean preoperative PIP, 35 +/- 10, mean postoperative PIP, 33 +/- 14 [P value not significant]). Six of the 8 nonsurvivors had NEC. Complications of intraabdominal abscess and enterocutaneous fistula were seen in 5 patients, all of who had NEC. CONCLUSIONS: Patch abdominoplasty effectively decreases airway pressures and oxygen requirements associated with ACS. Complications with PA occur primarily in patients with NEC. Failure to respond with a decrease in PIP and FIO2 requirements is an ominous sign. PMID- 10813331 TI - Potentially lethal complications of central venous catheter placement. AB - BACKGROUND: Placement of central venous catheters, although often considered to be a relatively safe and "junior"-level procedure, may be associated with life threatening complications. METHODS: A recent surgical death associated with placement of a central venous catheter at this Institution led to submission of a questionnaire to pediatric surgeons referenced through the American Pediatric Surgical Association directory regarding knowledge of similar incidents and information regarding catheter placement-related complications. RESULTS: Results to this response, although anecdotal, provided data regarding complications of an acute nature, which fell into the categories of pneumothorax, hydrothorax, cardiac tamponade, and hemothorax. Of 10 children with cardiac tamponade, 7 were infants, and most complications were associated with needle stick for access, with symptoms developing within minutes up to 12 hours after the procedure. Drainage of the tamponade was performed by aspiration alone in 3 cases; surgical drainage in 6 children resulted in survival in 9 of the 10 patients. Hemothorax was described in 19 patients and appeared to be more common in children in the 1- to 6-year age group, usually associated with percutaneous access techniques. Thoracotomy for hemothorax was performed in 16 children with 11 survivors. Vascular injury to subclavian artery, vein, or superior vena caval were noted in most at operation. CONCLUSIONS: Although data included in this review are entirely anecdotal and not subject to scientific scrutiny or analysis, certain conclusions appear evident. Inherent risks of central venous catheters are intrinsic and should be discussed with the family in obtaining preoperative consent, including life-threatening risks that may necessitate urgent surgical intervention (by thoracotomy or other means). Certain technical aspects of the procedure should be rigidly followed with an experienced surgeon in attendance throughout the procedure. Rapid evaluation should be performed for any unexplained problems that occur in the operating theatre or during the early postoperative period. PMID- 10813332 TI - Renal artery stenosis and pheochromocytoma: coexistence and treatment. AB - PURPOSE: The aim of this study was to develop an optimal treatment plan for coexisting pheochromocytoma and renal artery stenosis. METHODS: A retrospective analysis of the evaluation and management of a hypertensive 14-year-old boy was conducted. RESULTS: Secondary causes of hypertension were excluded initially including intracranial, cardiac, and endocrine abnormalities. A renal scan showed right renal function at 39%. Angiography confirmed a 90% subsegmental stenosis, as well as noting a blush suggesting a tumor. A computerized tomography (CT) scan of the abdomen showed a right adrenal mass. Serum epinephrine was 3,006 pg/mL (normal, <1,009 pg/mL) and 24-hour urinary norepinephrine was 2,001 microg (normal, <105 microg/24 h), suggesting the presence of a pheochromocytoma. During the operation for resection, a right subsegmental renal artery was found to be draped around a renal hilar mass; the adrenal gland was normal. The tumor was resected, and pathology confirmed a pheochromocytoma. Postoperatively, the patient had some mild hypertension that gradually resolved. CONCLUSIONS: Extraadrenal pheochromocytomas (paragangliomas) occur more frequently in children and are most commonly located in the renal hilum. In this location, they may present initially as renal artery stenosis as a result of direct arterial compression, fibrous bands, or catecholamine-induced vasospasm. Our experience supports tumor resection of extraadrenal pheochromocytomas as the definitive treatment for both conditions. PMID- 10813333 TI - Past and future of biliary atresia. AB - BACKGROUND: With the advent of liver transplantation the outcome of children with biliary atresia (BA) has improved. Is Kasai hepatic portoenterostomy (KHPE) still a valuable option for the treatment of these patients? METHODS: From 1974 to 1998, 77 patients with biliary atresia have been treated at our institution: 50 girls and 27 boys. RESULTS: Seventy-four patients had a KHPE, and 3 patients had no KHPE because of delay in diagnosis. A total of 65 of 74 patients (88%) had undergone KHPE type I, 4 patients (5.4%) KHPE type II, 3 patients (4%) had a Suruga modification, and 2 patients (2.6%) had a portocholecystostomy. Among the 74 patients, 11 were lost to follow-up and their cases were considered failures. Seventeen of our patients are alive at long-term follow-up after KHPE. Among the 77 patients, 33 (43%) had an orthotopic liver transplantation (OLT). Successful KHPE patients underwent transplant at a mean age of 9 years, and KHPE failed at a mean age of 11 months. A total of 25 of 77 (32%) of patients are alive thanks to OLT. In the cohort, the overall survival rate for the KHPE plus OLT is 42 of 77 (55%). Mortality and morbidity rates were more frequent among the younger patients who had early OLT after KHPE failure. In our series, overall survival rate was improved when the patient had a successful KHPE (P < .001). CONCLUSIONS: Kasai hepatic portoenterostomy (KHPE) continues to be a valuable procedure in the treatment of infants with biliary atresia (BA). Successful KHPE permits transplantation at an age at which mortality and morbidity are decreased leading to a better outcome. PMID- 10813334 TI - Fetus-in-fetu presenting as cystic meconium peritonitis: diagnosis, pathology, and surgical management. AB - Fetus-in-fetu (FIF), a rare congenital anomaly, is a fetus incorporating the well differentiated tissue of its twin. The authors describe a newborn who presented with massive abdominal distension and severe respiratory distress. Abdominal x rays showed multiple calcifications. The diagnosis of meconium pseudocyst was made. At emergency laparotomy an irregular fetiform mass was found in the retroperitoneum lying within a fluid-filled amniotic sac. It contained a vertebral column, 10 limblike structures, and cranial and caudal ends, supporting the diagnosis of fetus-in-fetu. This case highlights several important points. FIF often is overlooked in the differential diagnosis of a newborn abdominal mass and, as in this case, may be confused with meconuim pseudocyst. FIF should be differentiated from a teratoma because of the latter's malignant potential. Because this diagnosis is not made until pathological analysis, all parts of the mass should be removed to prevent malignant recurrence. PMID- 10813335 TI - Pelvic neuroblastoma: low mortality and high morbidity. AB - BACKGROUND/PURPOSE: Cervical, thoracic, and pelvic neuroblastomas are regarded as having a better outcome than abdominal primaries. The aim of the study was to analyze the results of treatment of pelvic neuroblastomas in our institution. METHODS: The authors reviewed the records of 284 patients with neuroblastoma treated in our hospital during the period 1983 through 1998 and identified 17 (6%) with pelvic tumors. The revised International Neuroblastoma Staging System was used. RESULTS: There were 6 patients with stage 1 disease, 8 with stage 2, 2 with stage 3, and 1 with stage 4 disease. Intraspinal extension of the tumor was present in 7 patients (41%). Except for one child with stage 4 disease, all patients underwent an attempt of tumor excision, and 6 had a complete resection. All 7 patients with intraspinal tumor survived. Permanent postoperative neurological complications occurred in 6 patients (35%). These included sciatic nerve palsy, urinary and fecal incontinence, neuropathic bladder, and leg weakness or nerve root injury L4-S1. Three of 17 patients died, but 1 fatality was unrelated to the tumor. The overall survival rate was 82% and was not influenced by the completeness of tumor resection. CONCLUSIONS: The survival of nonmetastatic pelvic neuroblastoma in our institution is good despite incomplete tumor resection. Intraspinal extension is not a negative prognostic factor. Considering the high incidence of permanent neurological damage after surgery and the generally favorable biological characteristics of these tumors, surgical treatment should not be overaggressive. PMID- 10813336 TI - The management of nonpigmented gallstones in children. AB - BACKGROUND/PURPOSE: An increasing number of children are diagnosed with gallstones today. The best management of nonpigmented gallstones in children without hematologic disorders is not known. METHODS: The authors prospectively studied 74 children with cholelithiasis diagnosed with ultrasonography. Clinical presentation, natural history, complications, and indications for cholecystectomy were examined. The follow-up (mean, 21 months) consisted of routine clinic visits, chart reviews, and telephone questionnaires with the children or their parents. RESULTS: Of the 74 children, 33 required cholecystectomies, and 41 were followed. The average age was similar in the 2 groups (11.7 v 11.0 years). Children with risk factors for cholelithiasis required earlier surgical treatment (P < .001). In the operative group, 8 presented acutely and 25 electively. There were 2 complications, a wound infection and a retained common duct stone. In the group that underwent follow-up, 34 of 41 children remained asymptomatic or had symptoms improve with dietary manipulation. No complications developed during the follow-up period. CONCLUSIONS: Children with gallstones and typical symptoms of right upper quadrant or epigastric pain with food intolerance should undergo cholecystectomy. Eighty-two percent of children with cholelithiasis and atypical symptoms had improvement with dietary manipulation. Pediatric patients with gallstones that are asymptomatic or associated with atypical symptoms can be safely followed without complications. PMID- 10813337 TI - Delayed presentation of a congenital recto-vaginal fistula associated with a recto-sigmoid tubular duplication and spinal cord and vertebral anomalies. AB - Tubular duplication of the recto-sigmoid colon is a rare entity. Associated anomalies including fistulae to the genitourinary tract may be found. A baby girl was found to have duplication of the recto-sigmoid colon, anomalies of sacral vertebra from S1 to S5, and solitary right kidney. The septum of this duplication was divided using staplers. Because of a history of stool coming from the vagina, a meticulous examination perioperatively was performed, but no fistula could be found. Further extensive investigation failed to show any fistula. At the age of 10 she was operated on for a tethered cord. At age 14, she experienced passage of a small amount of liquid stool per vaginum. A recto-vaginal fistula was found. Via a posterior sagittal incision, the fistula was closed by a transrectal approach. She remained asymptomatic for 16 months until the fistula recurred. Using a perineal approach, a very short fistula between the vagina and the rectum was closed. The closure was reinforced by a vaginal flap. Four months later, she remains without signs of recurrence. PMID- 10813338 TI - Amino acids counteract the inhibitory effect of fentanyl on hepatocyte oxidative metabolism. AB - BACKGROUND/PURPOSE: Hypothermia is common after major surgery in newborns and can be triggered by intraoperative fentanyl analgesia. Recent studies have found that fentanyl inhibits hepatocyte mitochondrial oxidative metabolism, which is proportional to thermogenesis. In adults it has been shown that amino acids have a thermogenic effect, although the biochemical basis of this phenomenon is not known. The aim of this study was to test the hypothesis that amino acids counteract the inhibition of neonatal hepatocyte oxygen consumption by fentanyl. METHODS: Hepatocytes were isolated from suckling rats, and O2 consumption was measured polarographically. In experiment A hepatocytes were incubated with (1) palmitate alone (control), (2) palmitate plus fentanyl, (3) palmitate plus fentanyl plus amino acids, and (4) palmitate plus amino acids. In experiment B the effects of essential and nonessential amino acids were tested separately. In experiment C, to investigate whether the effect of amino acids is intramitochondrial, hepatocytes were incubated with amino acids plus inhibitors of mitochondrial respiration. RESULTS: In experiment A, fentanyl significantly inhibited O2 consumption (P = .006). This inhibition was reversed by amino acids (P < .001). In experiment B, both essential and nonessential amino acids reversed the effect of fentanyl (P < .001). In experiment C, there was no difference in O2 consumption in the presence of myxothiazol among the groups indicating that amino acids affect intramitochondrial O2 consumption. CONCLUSIONS: (1) Amino acids abolish the inhibitory effect of fentanyl on hepatocyte oxidative metabolism. (2) Amino acids affect intramitochondrial O2 consumption and therefore thermogenesis. (3) Perioperative administration of amino acids in neonates may help to prevent hypothermia and its deleterious effects. PMID- 10813339 TI - Gastric motility disorders in patients operated on for esophageal atresia and tracheoesophageal fistula: long-term evaluation. AB - BACKGROUND/PURPOSE: Disturbed peristalsis is reported frequently after successful repair of esophageal atresia (EA). Delayed gastric emptying could be considered a cause of symptoms of gastroesophageal reflux (GER) in patients with repaired EA. The aim of the current study was to evaluate the incidence of and to characterize gastric motility disorders in a long-term follow-up of patients operated on for EA-tracheoesophageal fistula (TEF) by studying gastric emptying with scintigraphic techniques and comparing the results with gastric manometric data. METHODS: Eleven patients, between 12 and 23 years of age (median, 17) operated on for EA-TEF between 1975 and 1985, were studied. The scinthigraphic study was undertaken using a standard solid meal. The manometric study was performed using a 2.3-mm probe with 3 solid-state transducers. RESULTS: Dysphagia was present in about 20% of patients. Dyspepsia was recorded in 40% of the patients. A pathological reflux was present in 2 patients. Delayed gastric emptying (T1/2 > 90') was present in 4 patients (36%). Manometric data showed alteration of gastric peristaltic activity in 5 patients (45%). CONCLUSIONS: Delayed gastric emptying is frequent in long-term follow-up of patients operated on for EA-TEF. In these patients antral hypomotility also is recorded manometrically. Abnormal gastric motility can be considered as an important factor predisposing to symptoms of GER. A thorough evaluation of gastric function is recommended in symptomatic patients after EA repair. PMID- 10813340 TI - Combined pure esophageal atresia, duodenal atresia, biliary atresia, and pancreatic ductal atresia: prenatal diagnostic features and review of the literature. AB - The authors recently managed a case of combined pure esophageal, duodenal, biliary, and pancreatic ductal atresia in a fetus and newborn with Trisomy 21. The authors present a case report and review of the literature, emphasizing the prenatal radiographic features of the combined lesions and the high incidence of associated anomalies including Down's syndrome. Prenatal suspicion of these anomalies warrants karyotype analysis as well as careful pre- and postnatal screening for other anomalies. PMID- 10813341 TI - Marked morphological differences in the myenteric plexus between the mesenteric and antimesenteric sides of small bowel in premature infants. AB - BACKGROUND: The gastrointestinal tract appears morphologically prepared for oral feeding by the end of the second trimester, but many of the physiological processes required for efficient enteral nutrition are not developed fully until 33 to 34 weeks' gestation. Myenteric plexus is well recognized as an important regulator of peristaltic activity. Whole-mount preparation technique produces a 3 dimensional picture to better show the neuronal networks branching and interconnections. The aim of this study was to investigate neurone density and morphology of the myenteric plexus in premature infants using whole-mount technique. METHODS: Full-thickness small and large bowel specimens were collected at autopsy from 6 premature babies (gestational age, 26 to 32 weeks) who died without evidence of gastrointestinal disease. Whole-mount preparation of the myenteric plexus was made and stained with NADPH-diaphorase and Acetylcholinesterase (AChE) histochemistry. The stained myenteric network was measured with a computer image analysis system. Controls included 4 full-term babies who died of nongastrointestinal disease. RESULTS: In premature infants there were striking differences in neuronal density of myenteric plexus in the mesenteric and antimesenteric border of small bowel. The differences in neuronal density in mesenteric and antimesenteric border of small bowel gradually became less striking as the gestation progressed with no differences evident at gestational age 32 weeks. CONCLUSIONS: This study shows for the first time that the neurone density of myenteric plexus is significantly higher in the mesenteric border of the small bowel compared with antimesenteric border in premature infants. The marked morphological differences observed in neurone density in the small bowel of premature infants may contribute to immature small bowel activity. PMID- 10813342 TI - Superior mesenteric venous thrombosis in malrotation with chronic volvulus. AB - Malrotation can be difficult to diagnose after the newborn period because of intermittent symptoms and vague clinical findings, but malrotation with midgut volvulus is usually quite striking in its presentation. Early diagnosis and surgical treatment are essential to prevent acute ischemic infarction of the bowel, although chronic complications are rare. The authors present an unusual case of mesenteric venous thrombosis secondary to chronic midgut volvulus. A 13 year-old girl presented with an 11-year history of recurrent bouts of abdominal pain evaluated at 3 other institutions without a diagnosis. At the referring hospital, an episode of bilious emesis associated with abdominal pain prompted a computerized tomography scan of the abdomen. This showed a calcified thrombus within the superior mesenteric vein (SMV). At laparotomy, malrotation with chronic 270 degree volvulus was found with evidence of mesenteric venous hypertension. Segmental occlusion was documented on magnetic resonance angiography. SMV thrombosis is an unusual complication of malrotation with chronic midgut volvulus. PMID- 10813343 TI - Should malrotation in children be treated differently according to age? AB - PURPOSE: The aim of this study was to better define the mode of presentation, rate of volvulus, and surgical findings in children younger than 2 versus older than 2 years of age with malrotation. METHODS: The authors reviewed the charts of all patients with malrotation admitted to their hospital between January 1980 and December 1998, excluding patients having malrotation as a secondary finding. RESULTS: An upper gastrointestinal series was done in 90 patients (6% falsely negative) and a barium enema in 20 patients (40% read as normal). Fifty-eight patients had 114 associated congenital anomalies. Volvulus was found at the time of surgery in 28 patients, 5 of whom were older than 2 years. Three presented with acute symptoms and 2 with chronic symptoms. Surgery was performed by laparotomy in 103 patients and by laparoscopy in 3. Mean length of stay was 13.6 days. Mean follow-up was 19 months. Death occurred in 4 patients; postoperative bowel obstruction was seen in 3 patients (only 1 required surgery). CONCLUSIONS: Children with malrotation who are older than 2 years old have a significant risk of volvulus that is difficult to predict radiologically. They require surgical attention even if asymptomatic. Laparoscopy allows evaluation of the base of the mesentery and completion of the Ladd's procedure. PMID- 10813344 TI - Intestinal metabolism after ischemia-reperfusion. AB - PURPOSE: This study explores the effects of ischemia-reperfusion on various metabolic aspects of the small intestine. METHODS: Intestinal ischemia reperfusion was obtained by clamping and unclamping the superior mesenteric artery in adult rats. Four groups of animals were studied: (A) sham operation for 150 minutes, (B) 90-minute intestinal ischemia, (C) 150-minute intestinal ischemia, and (D) 90-minute intestinal ischemia followed by 60-minute reperfusion. Body temperature was maintained at normothermia (36.5 to 37.5 degrees C). Concentrations of intestinal glucose, succinate, lactate, amino acids, phosphocholine (PC), glycerophosphocholine (GPC), choline, and phosphoenergetics were measured using magnetic resonance spectroscopy of freeze clamped small intestine extracts. RESULTS: Intestinal ischemia (groups B and C) alone caused a significant drop in glucose and phosphoenergetics but caused an increase in amino acids, succinate, and lactate. Ischemia and ischemia reperfusion decreased PC and GPC but increased choline. After intestinal reperfusion (group D), no recovery of phosphoenergetics was observed, but there was partial recovery of glucose, succinate, lactate, and amino acids. CONCLUSIONS: There is no recovery of phosphoenergetics after 90 minutes of intestinal ischemia followed by 60 minutes of reperfusion. Partial recovery of glucose, succinate, lactate, and amino acids may reflect equilibration of these metabolites between damaged cells and extracellular fluid. PMID- 10813345 TI - Breast cancer in a 6-year-old child. AB - A 6-year-old girl presented for a second opinion with a 1-year history of an enlarging soft tissue mass just lateral to the right areola. She had been seen by a pediatric surgeon elsewhere who reassured the parents that the lesion was benign. Ultrasound scan showed a 1.5- x 1.5-cm cystic structure adjacent to the right breast bud. Excisional biopsy results showed secretory ductal adenocarcinoma. Modified radical mastectomy with axillary node dissection was performed. All 11 nodes were negative for metastatic disease. She is now disease free 3 years after diagnosis. Estrogen-progesterone receptors were negative, as was screening for BCR 1 and 2. This is the first report of cytogenetics showing an abnormal cell line with a reciprocal translocation between 12p and 15q. Although breast cancer is extremely rare in children, a history of a painless, enlarging, firm breast mass should raise concern about possible neoplastic disease. Cystic appearance on ultrasound scan caused by the pseudocapsule around the tumor may be a marker for secretory carcinoma. Histological evaluation of all suspicious masses should be obtained. Because of the risk of local recurrence and axillary metastases, the authors recommend modified radical mastectomy with axillary node dissection for children with secretory carcinoma of the breast. PMID- 10813346 TI - Thoracoscopic thymectomy in juvenile myasthenia gravis. AB - BACKGROUND: Although transsternal thymectomy is an effective method in the treatment of juvenile myasthenia gravis (JMG) it is traumatic in pediatric patients. Thoracoscopic thymectomy offers an effective and less traumatic approach with respect to cosmesis and postoperative recovery. METHODS: A retrospective analysis of 6 consecutive patients treated with thoracoscopic thymectomy was performed. Perioperative parameters and cost analysis were compared with those of 6 consecutive open procedures performed before the study. RESULTS: Thoracoscopic thymectomy can be performed in patients as young as 1.6 years. There was no conversion to open procedure and no perioperative morbidity and mortality. The length of operating time and the surgical cost of thoracoscopic procedure were not significantly different from those of open procedure. The length of hospitalization, however, was significantly shorter with thoracoscopic procedure, and hence the overall cost was significantly reduced (P < .05). An intermediate follow-up shows that outcome after thoracoscopic procedure is equally as effective as open procedure. CONCLUSIONS: Thoracoscopic thymectomy offers an equally effective but cosmetically more acceptable approach than sternotomy. It has a quicker recovery period and appears to be a less costly alternative to transsternal thymectomy. PMID- 10813347 TI - Leiomyoma of the esophagus associated with bronchial obstruction owing to inflammatory pseudotumor in a child. AB - Although relatively common in adults, leiomyoma of the esophagus is a rare disorder in children. A single case report describes the coexistence of both esophageal and bronchial leiomyoma in a child. The authors describe the diagnostic and treatment challenges encountered in a 2-year-old boy with coexisting inflammatory pseudotumor and esophageal leiomyoma presenting as massive atelectasis. PMID- 10813348 TI - Short-term tracheal occlusion in fetal lambs with diaphragmatic hernia improves lung function, even in the absence of lung growth. AB - BACKGROUND/PURPOSE: Prolonged tracheal occlusion (TO) accelerates lung growth but impairs surfactant production. Short-term TO results in less lung growth but preserves type II cell function. The authors studied the effects of short-term TO on lung physiology in diaphragmatic hernia. METHODS: Diaphragmatic hernia was created in 9 fetal lambs at 90 to 95 days. Five were left uncorrected (CDH), 4 underwent 2-week TO (108 to 122 days; CDH + TO). Five unoperated lambs served as controls. Near-term (136 days) fetuses were ventilated for 90 to 150 minutes. Pulmonary arterial pressure, postductal blood gases, quasistatic compliance, total lung capacity (TLC), and lung weight to body weight (LW/BW) were measured. RESULTS: There was an overall survival rate of 89% at full term. Short-term occlusion did not induce lung growth (TLC and LW/BW, 6.07 +/- 2.92 mL/kg and 0.022 +/- 0.008 in CDH, 4.86 mL/kg and 0.019 +/- 0.005 in CDH + TO, 10.81 +/- 3.55 mL/kg and 0.036 +/- 0.006 in controls, respectively). However, pulmonary hypertension in CDH (47.4 +/- 12.32/35.8 +/- 12.19 torr) was corrected by short term occlusion (20.2 +/- 4.0/16.0 +/- 4.8 torr in CDH + TO, P< .05, single-factor analysis of variance [ANOVA]; similar to control). Best pO2 and pCO2 improved after occlusion (CDH, 48.6 +/- 6.7 torr and 107.1 +/- 34.3 torr, respectively; CDH + TO, 101.5 +/- 16.3 torr and 81.9 +/- 2.4 torr; control, 291.4 +/- 4.7 torr and 37.7 +/- 17.3), as did oxygenation index (P < .05, CDH vCDH + TO; CDH, 97.2 +/- 23.0; CDH + TO, 28.7 +/- 3.1; control, 5.6 +/- 0.6). CONCLUSIONS: Short-term TO corrects pulmonary hypertension and improves gas exchange in fetal lambs with diaphragmatic hernia despite failure to produce accelerated lung growth. Inducing lung maturation and correcting the physiological derangement in diaphragmatic hernia may be more important than achieving lung growth alone. PMID- 10813349 TI - Effect of CGS 26303, an endothelin-converting enzyme-neutral endopeptidase inhibitor, on nitrofen-induced congenital diaphragmatic hernia in the rat. AB - BACKGROUND/PURPOSE: The pathophysiology of congenital diaphragmatic hernia (CDH) associated with lung hypoplasia and pulmonary hypertension is not understood fully. Endothelins (ETs) are the most potent vasoconstrictors that also act as promitogenic agents. They may play a role during pregnancy in leading to the condition found at birth and ongoing mortality in CDH. Therefore, the authors studied the effect of CGS 26303, a nonselective endothelin-converting enzyme and neutral endopeptidase inhibitor, in the rat model of CDH. METHODS: Pregnant Sprague-Dawley rats were divided into 3 groups: group 1 (n = 4) received CGS 26303 (50 mg/kg, subcutaneously, twice a day), from gestational day 12 until term (21 to 23 days); group 2 (n = 8) received nitrofen (100 mg/kg, orally) at gestational day 11.5; group 3 (n = 8) received both nitrofen and CGS 26303. The survival of the newborn rats was monitored up to 240 minutes. After natural death or euthanasia, they were weighed and microdissected. The degree of hernia was quantified as small, moderate, or severe, and lungs and liver were harvested and weighed. RESULTS: Newborn rats from mothers of group 3 (n = 81) survived 196 +/- 8 minutes compared with 173 +/- 9 minutes of those of group 2 (n = 97). Severe CDH from group 3 (n = 20) had a mean survival time of 66 +/- 13 minutes compared with 26 +/- 4 minutes for those of group 2 (n = 27). Lung index in severe CDH pups of group 3 was increased by 13% compared with those from group 2 (P < .0001), whereas their liver index went down by 8% (P < .05). CONCLUSIONS: These results suggest that CGS 26303 might have a beneficial effect when given during pregnancy in increasing survival at birth and reducing the severity of the pulmonary hypoplasia in newborn rats with nitrofen-induced CDH. PMID- 10813350 TI - Life-threatening air rifle injuries to the heart in three boys. AB - Air rifles, or BB guns, are generally thought of as childhood toys. Although most injuries are not serious, life-threatening events have been reported. Within a 1 year period, 3 boys presented after BB gun shots to the chest, all requiring surgical intervention for penetrating injuries to the heart. A 15-year-old underwent window pericardiotomy for hemopericardium with thrombus 24 hours after admission. Another, 5 years of age, underwent emergent exclusion of the cardiac apex for a traumatic ventricular septal defect. The third, 8 years old, had a right ventricular injury requiring an urgent subxiphoid pericardial window for tamponade. All recovered uneventfully. Increased public awareness, adult supervision, safety training, and appropriate legislation are needed to decrease the risks of these potentially lethal weapons. PMID- 10813351 TI - Complications of splenic injuries: expansion of the nonoperative theorem. AB - BACKGROUND/PURPOSE: Delayed complications (hemorrhages, abscesses, and pseudo aneurysms) of nonoperative management (NOM) in pediatric spleen injuries are rare but reportedly result in failure of NOM. This study was undertaken to elucidate the rate of delayed complications and their impact on NOM of splenic injuries. METHODS: Forty children who underwent NOM of splenic injuries over 5 years were reviewed for complications and success of NOM. All injuries were diagnosed and graded by computed tomography (CT) scans. Follow-up imaging studies were obtained in all children 6 to 12 weeks postinjury; additional scans also were obtained when clinically indicated. RESULTS: NOM was successful in all children, but 3 (7.5%) had delayed complications. A 3 year old with a grade III splenic injury and a distal pancreatic transection, and a 13 year old with a grade IV injury had pseudoaneurysms, which were noted on follow-up CT scan (14 days postinjury). The pseudoaneurysms were treated expectantly and resolved spontaneously. A 16 year old (grade IV injury) had a splenic abscess 8 days postinjury that was drained percutaneously with CT guidance. CONCLUSIONS: Development of delayed complications may not preclude successful NOM of pediatric spleen injuries. Splenic artery pseudoaneurysms in children appear to resolve spontaneously without intervention. PMID- 10813352 TI - Prenatal diagnosis and management of congenital lobar emphysema. AB - BACKGROUND: Congenital lobar emphysema (CLE) is a rare anomaly of lung development that usually presents in the neonatal period with respiratory distress and pulmonary lobar hyperinflation. The routine use of prenatal ultrasonography has resulted in the early identification and serial evaluation of congenital lung lesions. CLE can be distinguished from other congenital lung lesions on ultrasonography by the differences in echogenicity and reflectivity. METHODS: Two cases of CLE diagnosed at midgestation by ultrasonography and ultrafast fetal magnetic resonance imaging (MRI), along with serial sonographic documentation of their prenatal course were reviewed. RESULTS: The CLE lesions decreased in size over the course of the pregnancy, similar to that seen with other congenital lung lesions such as cystic adenomatoid malformation and bronchopulmonary sequestration. However, these neonates with CLE showed marked air-trapping and respiratory distress requiring lobectomy in the early neonatal period. CONCLUSIONS: These cases provide insight into the prenatal course of CLE and underscore the need for continued postnatal evaluation of fetuses even those in whom the lesions appear to have resolved in utero. These patients should have ready access to postnatal surgical intervention. PMID- 10813353 TI - Pulmonary lymphangiectasia revisited. AB - PURPOSE: Pulmonary lymphangiectasia (PL) is a rare, poorly documented disease characterized by abnormal pulmonary lymphatics. Although case reports are published, little is known about survivors past the neonatal period. METHODS: This is a retrospective review of histologically proven PL in fetuses, infants, and long term survivors since 1965. RESULTS: Eleven children (8 boys, 3 girls) and 8 aborted fetuses (7 male, 1 female) were identified. The fetuses weighed 463.4 g (177 to 681 g). Six were aborted between 19 to 24 weeks of gestation for multiple malformations or anencephaly, and 2 spontaneously aborted: one with PL only, the other with twin-twin transfusion syndrome. Clinical PL was diagnosed between 0 and 11 months of age. Six children died (2 neonatal, 4 within 10 days), 5 survived. Two deaths occurred after cardiac surgery. Among survivors, the symptomatology and frequency of admissions diminished over time. Symptoms included progressive respiratory distress, chronic cough, recurrent pneumonia, bronchial asthma, and choking. One child with bilateral chylothorax was later diagnosed with Noonan syndrome; 2 patients had minor cardiac malformations. Rapid deterioration occurred with mild respiratory infections with only supportive treatment available. Chest x-ray showed marked hyperinflation with interstitial infiltrate. CONCLUSIONS: This is the first long-term study of primary PL and will help counsel parents. Although fatal in the neonatal period, survival is possible if diagnosed past the neonatal period and improvement is expected. PMID- 10813354 TI - The unpredictable character of congenital cystic lung lesions. AB - BACKGROUND: The spectrum of congenital cystic disease of the lung ranges from hydrops and neonatal respiratory distress to asymptomatic lesions. Surgical management is dictated by the presence of symptoms, recurrent infection, and the potential risk of malignant transformation. METHODS: Since 1995, all consecutive patients with congenital cystic lung lesions underwent follow-up for symptoms, treatment, and correlation of presumptive with pathological diagnosis. RESULTS: Twelve cystic lung lesions were identified. Seven were diagnosed with mediastinal shift in utero; in 6 of 7, the shift subsequently resolved. Overall, 6 of 7 lesions that were followed up serially decreased in size. Two patients were symptomatic in utero; 1 underwent thoracoamniotic shunting, 1 pleurocentesis for impending hydrops. Postnatally, these 2, and 2 other newborns required urgent surgery. Five of 8 asymptomatic patients had elective resection by 16 months, and 4 await operation. In 6 of the 9 surgical cases (67%), there was a discrepancy between preoperative and pathological diagnosis. There were 4 hybrid congenital cystic adenomatoid malformation (CCAM)/sequestrations. CONCLUSIONS: At least 6 of 7 congenital cystic lung lesions decreased in size regardless of gestational age or presence of mediastinal shift. Antenatal intervention is therefore rarely indicated. Hybrid morphology may necessitate resection of stable, asymptomatic lesions to prevent the theoretical concern for associated malignancies as well as other complications of CCAM. PMID- 10813355 TI - An infant with a cervical injury sometimes requires stabilization with a halo while surgery or the injury heals. PMID- 10813356 TI - Expert witness testimony in medical liability cases: American Pediatric Surgical Association position statement. PMID- 10813357 TI - Childhood intussusception: a comparative study of nonsurgical management. PMID- 10813358 TI - Ulceration in an ileocolic anastomosis treated with Ranitidin. PMID- 10813359 TI - Anomalous arrangement of the pancreaticobiliary duct (AAPBD) PMID- 10813360 TI - Expression of the lung-specific thyroid transcription factor (TTF-1) within the tracheoesophageal fistula of embryo rats exposed to Adriamycin. PMID- 10813361 TI - Gramicidin toxicity in NG108-15 cells: protective effects of acetamidine and guanidine. AB - Studies were conducted using a novel in vitro approach to investigate the efficacy of acetamidine hydrochloride (ACE) and guanidine hydrochloride (GUAN), previously shown to block gramicidin D (GRAM) channels in artificial membranes, in preventing the toxic effects of GRAM in NG108-15 (neuroblastoma x glioma hybrid) cells. Specifically, intracellular microelectrode techniques were employed to examine changes in membrane resting potential (Vm) and input resistance (Rin). At 1 micromol/L, ACE significantly reduced loss of Vm induced by 1 or 10 microg/ml GRAM, although higher concentrations of ACE did not afford enhanced antagonism. GUAN, in contrast, produced a concentration-dependent antagonism of GRAM-induced Vm and Rin loss, with high concentrations (10 or 100 micromol/L) completely preventing diminutions in both Vm and Rin. In control cells superfused without GRAM, ACE produced a direct, concentration-dependent reduction in Vm and Rin, whereas GUAN hyperpolarized NG108-15 cells but did not alter Rin. These data represent the initial demonstration of the reversal of GRAM toxicity in an intact cell system. PMID- 10813362 TI - The SOS induction of umu'-'lacZ fusion gene by oxidative damage is influenced by polyamines in Escherichia coli. AB - We report that polyamines have an effect on the SOS response of the umu operon in polyamine-deficient mutant and wild-type Escherichia coli strains carrying the umu'-'lacZ fusion. H2O2 effectively induces umu'-'lacZ in the wild type, but not significantly in the mutant strain. Exogenous polyamines did not restore the umu induction in the mutant to the wild-type level. In logarithmically growing cells, the basal expression of umu gene in the mutant is about five times higher than that of the wild type. The addition of polyamines to the growth medium markedly reduces the basal expression to the wild-type level. This reduction is due not to growth rate but to the polyamine itself. Our results suggest that polyamines are essentially involved in the SOS induction of the umu operon in E. coli. PMID- 10813363 TI - Development and characterization of a cell line from Pacific herring, Clupea harengus pallasi, sensitive to both naphthalene cytotoxicity and infection by viral hemorrhagic septicemia virus. AB - A cell line, PHL, has been successfully established from newly hatched herring larvae. The cells are maintained in growth medium consisting of Leibovitz's L-15 supplemented with 15% fetal bovine serum (FBS), and have been cryopreserved and maintain viability after thawing. These cells retain a diploid karotype after 65 population doublings. PHL are susceptible to infection by the North American strain of viral hemorrhagic septicemia (VHS) virus, and are sensitive to the cytotoxic effects of naphthalene, a common environmental contaminant. Naphthalene is a component of crude and refined oil, and may be found in the marine environment following acute events such as oil spills. In addition, chronic sources of naphthalene contamination include offshore drilling and petroleum contamination from areas such as docks and marinas that have creosote-treated docks and pilings and also receive constant small inputs of petroleum products. This cell line should be useful for investigations of the toxicity of naphthalene and other petroleum components to juvenile herring. In addition, studies of the VHS virus will be facilitated by the availability of a susceptible cell line from an alternative species. PMID- 10813364 TI - Clofibrate inhibits membrane trafficking to the Golgi complex and induces its retrograde movement to the endoplasmic reticulum. AB - Insights into the function of the Golgi complex have been provided by experiments performed with various inhibitors of membrane trafficking, such as the macrocyclic lactone brefeldin A (BFA), a compound that inhibits constitutive secretion, prevents the formation of coatomer-coated transport vesicles, and stimulates the retrograde movement of Golgi resident enzymes back to the ER. We show here that the structurally unrelated compound clofibrate, a peroxisome proliferator (PP) and hypolipidemic agent, also reversibly disrupts the morphological and functional integrity of the Golgi complex in a manner similar to BFA. In the presence of clofibrate, the forward transport of newly synthesized secretory proteins from the ER to the Golgi is dramatically inhibited. Moreover, clofibrate causes Golgi membranes to travel rapidly in a microtubule-dependent manner back to the ER, forming a hybrid ER-Golgi tubulovesicular membrane network. These affects appear to be independent of clofibrate's ability to stimulate the PP-activated receptor (PPAR) alpha pathway because other PPAR stimulators (DEHP, WY-14643) did not alter the Golgi complex or induce retrograde trafficking. These data suggest that PPAR alpha-independent, clofibrate-sensitive proteins participate in regulating Golgi-to-ER retrograde membrane transport, and, equally importantly, that clofibrate may be used as a pharmacological tool for investigating Golgi membrane dynamics. PMID- 10813365 TI - Glutathione-dependent cytotoxicity of the chloroacetanilide herbicides alachlor, metolachlor, and propachlor in rat and human hepatoma-derived cultured cells. AB - Alachlor, metolachlor, and propachlor are widely used chloroacetanilide herbicides. Their cytotoxicity in rat (Fa32) and human (Hep G2) hepatoma-derived cells was investigated, in connection with their influence on the endogenous glutathione (GSH) content, on the xenobiotic-metabolizing phase I enzymes 7 ethoxyresorufin O-deethylase (EROD) and 7-pentoxyresorufin O-depentylase (PROD), and phase II glutathione transferase (GST). The cytotoxicity was measured by the neutral red uptake inhibition assay. The following toxicity range was observed in both cell lines: propachlor > alachlor > metolachlor. When the endogenous GSH content was reduced by pretreatment of the cells with L-buthionine (S,R) sulfoximine, the cytotoxicity of the herbicides increased strongly in both cell lines. EROD and PROD activities were dose-dependently increased to different degrees in Fa32, as was EROD in Hep G2, but no PROD activity was observed in these cells. The GSH content was not altered after 1 h treatment, and was approximately doubled after 24 h. GST activity was increased in Fa32 cells but not in Hep G2. A comparable cytotoxicity was observed for the investigated chloroacetanilides in both the rat and the human cell lines. Different interactions with xenobiotic-metabolizing phase I and II enzymes were observed, and GSH showed a protective effect against the acetanilides in both cell lines. PMID- 10813366 TI - Effect of toluene diisocyanate and its corresponding amines on viability and growth of human lung fibroblasts in culture. AB - Toluene diisocyanate (TDI) is a highly volatile chemical known to cause occupational asthma in exposed workers. TDI-induced asthma is associated with airway epithelium injury and repair, and subepithelial fibrosis. We investigated the effect of TDI and its hydrolysis products, the 2,4- and 2,6-toluenediamines (TDA), on viability and growth of human lung fibroblasts (HLFs) in culture, using a tetrazolium-based cell viability assay. The effects of increasing concentrations of each of these chemicals were evaluated on quiescent cells seeded at two densities (2500 and 5000 cells/well) and treated for 24 or 48 h. TDI (10(-4)-10(-5) mol/L, as a mixture of 80% 2,4-TDI and 20% 2,6-TDI) exhibited a partial but significant cytotoxic effect (10-24%, p<0.05) on HLFs. This effect was observed at both cell densities, and was time- and concentration-dependent. 2,4-TDA, at lower concentrations (10(-8)-10(-6) mol/L) applied for 48 h, also partially reduced HLF viability (10-15%, p<0.05), whereas it tended to trigger cell growth at concentrations higher than 10(-5) mol/L. 2,6-TDA exhibited both a cytotoxic and a proliferative effect on HLFs that depended on concentration, time of exposure and cell culture density. Significant cytotoxicity was only observed after 24 h of treatment with 10(-7)-10(-6) mol/L 2,6-TDA, and reached greater intensity in cells cultured at the highest density. In contrast, 2,6-TDA stimulated HLF growth only after 48 h of incubation at 10(-4) mol/L on cells cultured at the lowest density. Taken together, our results showed that TDI and 2,4-TDA somewhat decreased HLF viability, whereas 2,6-TDA appeared to exhibit both a cytotoxic and a growth stimulatory effect on these cells. TDI and 2,4-TDA are thus suggested to contribute to airway epithelium damage associated with TDI induced asthma, whereas 2,6-TDA might either trigger epithelial damage or induce cell proliferation that could contribute to epithelium repair or subepithelial fibrosis. PMID- 10813367 TI - ET-receptor subtypes: roles in regional renal vascular actions of exogenous and endogenous endothelins in anesthetized rabbits. AB - The roles of endothelin (ET)-receptor subtypes, in the regional renal vascular effects of exogenous and endogenous ETs, were examined in pentobarbitone anesthetized rabbits. The effects of renal arterial infusion of ET-1 (0.05-12.8 ng/kg/min) and the ET(B)-agonist [Ala1,3,11,15]-ET-1 (12.5-800 ng/kg/min) were compared. We then tested the effects of the ET(A)-antagonist BQ610 and the ET(B) antagonist BQ788 (both 200 microg/kg plus 100 microg/kg/h, i.v.) on basal hemodynamics and on responses to renal arterial ET-1. Both ET-1 and [Ala1,3,11,15]-ET-1 dose-dependently reduced total renal blood flow (RBF) and cortical blood flow (CBF), but not medullary blood flow (MBF). ET-1 was 34-fold more potent than [Ala1,3,11,15-ET-1. BQ610 reduced mean arterial pressure (MAP; 14%), and increased RBF (21%) and CBF (12%), but not MBF. BQ788 increased MAP (13%), and reduced RBF (29%) and CBF (15%) but not MBF. Coadministration of both agents increased RBF (18%) and CBF (9%), without significantly affecting MAP. Neither antagonist (alone or combined) significantly affected responses to renal arterial ET-1. We conclude that the predominant renal vascular effects of exogenous and endogenous ETs are cortical vasoconstriction, but not at vascular sites controlling MBF. ET(A)-receptors contribute to the renal vasoconstrictor effects of endogenous ETs. ET(B2)-like receptors appear to contribute to the vasoconstrictor effects of [Ala1,3,11,15]-ET-1. PMID- 10813368 TI - Effect of reserpine treatment on low-density lipoproteins in arterial wall and internal organs of rats. AB - The effects were determined in rats of single injections of reserpine at increasing doses (0.5, 1.58, and 5.0 mg/kg) on low-density lipoprotein (LDL) and cholesterol in aortic wall, heart, liver, kidney, and adrenal gland. Catecholamine levels in plasma, heart, and liver, arterial blood pressure, and heart rate were also monitored. Reserpine was injected intraperitoneally, followed immediately by the administration of [3H]cholesterol by gavage. Twelve hours later, homologous 125I-tyramine cellobiose-labeled LDL (125I-TC-LDL) was injected intravenously. Twenty-four hours later, the rats were killed, and the radioactivities of aortic walls, heart, liver, kidney, and adrenal glands were determined. The results showed that after reserpine treatment the accumulation of both the 125I-TC label derived from LDL and total [3H]cholesterol was significantly reduced in aortic wall and heart, increased in liver, and unchanged in the kidney and adrenal gland. At higher doses (1.58 and 5.0 mg/kg), reserpine significantly accelerated the plasma clearance of radiolabelled LDL. Plasma noradrenaline in reserpine-treated animals decreased maximally (86%) by 12 h and by 61-71% at 36 h compared with the control. Plasma adrenaline increased transiently after injection of reserpine and then returned to the basal levels. Reserpine greatly decreased noradrenaline and adrenaline levels in heart and liver. Arterial blood pressure was decreased significantly (0.001 < p < 0.05) at 12 h by the two lower doses of reserpine and then returned to normal values over the next 24 h. The results indicate that reserpine decreases LDL cholesterol in artery wall and heart and increases it in liver. These findings suggest that reserpine could find a new use as a cholesterol-lowering drug for the prevention of atherosclerosis. PMID- 10813369 TI - Evidence that nitric oxide regulates AT1-receptor agonist and antagonist efficacy in rat injured carotid artery. AB - Vascular injury stimulates AT1-receptor expression and nitric oxide (NO) production in smooth muscle cells (SMCs). We examined the ability of AT1 agonists and antagonists to regulate vascular tone ex vivo in injured arteries and the possible modulation by SMC-derived NO. Rings of rat carotid arteries were isolated at day 7 after endothelial denudation and stimulated with angiotensin (Ang) II in the absence or presence of the AT1 antagonists losartan, L-158,809, or EXP-3174. Freshly denuded contralateral arteries were used as controls. AngII induced contractions were similar in control and injured arteries. Losartan caused an insurmountable inhibition of AngII-induced contractions in injured but not control arteries. Enhanced inhibition of AngII in injured arteries also was observed in the presence of L-158,809 and EXP-3174. In the presence of the NO synthesis inhibitor nitromonomethyl-L-arginine (L-NMMA), maximal contractions to AngII were greater in injured than in control vessels, and AT1-receptor blockade with losartan was surmountable in all vessels. Mechanical removal of superficial neointimal SMCs attenuated NO production and normalized the efficacy of losartan in injured arteries. These results suggest a role for NO in reducing the biologic effects of AT1-receptor agonists and potentiating the efficacy of AT1 antagonists in vessels undergoing remodeling after injury. PMID- 10813370 TI - Crataegus special extract WS 1442 increases force of contraction in human myocardium cAMP-independently. AB - The mode of action of Crataegus extracts in the treatment of heart failure is still under examination. WS 1442, a standardized special extract from Crataegus leaves with flowers, exerts direct positive inotropic effects. This study was designed to investigate the mode of inotropic action of WS 1442 in human myocardium from patients with congestive heart failure (left ventricular myocardium from explanted hearts; NYHA IV, n = 8) as well as in nonfailing controls (right auricular trabeculae from patients with coronary heart disease, n = 8). WS 1442 effectively displaced specifically bound 3H-ouabain but did not influence the activity of adenylate cyclase [control, + Gpp(NH)p (10(-4) microM) 3,500 pmol cyclic adenosine monophosphate (cAMP)/20 min). In isolated left ventricular papillary muscle strips, WS 1442 significantly increased the force of contraction [basal, 1.8+/-0.2 mN; WS 1442 (50 microg/ml), 2.4+/-0.1 mN (130%)] and improved the frequency-dependent force generation (0.5 vs. 2.5 Hz: control, +0.1+/-0.01 mN; WS 1442, +0.9+/-0.3 mN) even in failing human myocardium. In fura 2-loaded muscle strips (right atrial trabeculae), WS 1442 increased both the Ca2+ transient and force generation. These effects also were observed in the lipophilic ethyl acetate-soluble fraction A, enriched in flavone derivatives. In conclusion, these findings suggest a pharmacologic mechanism of WS 1442 similar to the cAMP-independent positive inotropic action of cardiac glycosides. In addition, WS 1442 improves the force-frequency relation in failing human myocardium. PMID- 10813371 TI - Oxygen radical system in chronic infarcted rat heart: the effect of combined beta blockade and ACE inhibition. AB - In vitro experiments suggest that beta blockade and angiotensin-converting enzyme (ACE) inhibition may protect the failing heart by reduction of myocardial oxidative stress. To test this hypothesis in an in vivo model, the beta blocker metoprolol (350 mg) and the ACE inhibitor ramipril (1 mg) were given either alone or in combination to rats (per kilogram body weight per day) for 6 weeks after myocardial infarction. Left ventricular end-diastolic pressure (LVEDP), contractile function of papillary muscles, enzymatic antioxidative defense (indicated by the activities of the superoxide dismutase isoenzymes and glutathione peroxidase), and the extent of lipid peroxidation were studied. Placebo-treated rats showed cardiac hypertrophy, increased LVEDP, lower rates of contraction and relaxation, as well as a deficit in the myocardial antioxidative defense associated with increased lipid peroxide levels, when compared with sham operated animals. Combined beta blockade and ACE inhibition improved the antioxidative defense, reduced hypertrophy and LVEDP, and enhanced rates of contraction. Thus prolonged beta blockade and ACE inhibition after infarction may decrease myocardial oxidative stress and thereby could be beneficial in heart failure. PMID- 10813372 TI - E 047/1: a new class III antiarrhythmic agent. AB - The efficacy, pharmacokinetics, safety, and tolerability of E 047/1, an amiodarone derivative, were evaluated in patients with acute supraventricular or ventricular arrhythmia. In an open, nonrandomized prospective multicenter trial, 20 patients were treated with three different i.v. dosage regimens of E 047/1. Arrhythmia termination indicated efficacy. Pharmacokinetics were determined by measurements of drug plasma levels. Safety was judged by changes of blood pressure, heart rate, ECG parameters, and appearance of adverse events. For local tolerability, effects at the site of infusion were assessed. In patients with atrial fibrillation and/or atrial flutter, drug plasma levels and prolongation of QT interval were correlated with efficacy. In 10 (50%) patients, therapeutic intervention with E 047/1 was successful. Drug plasma levels rapidly decreased within 1 h after administration. Blood pressure values and ECG parameters stayed constant during the observation period. Proarrhythmic effects were not observed. As adverse events, vertigo, vomiting, and nausea in three (15%) and hypotension in one (5%) patient, respectively, occurred in the high-dose bolus regimen only. At the site of infusion, no adverse effects were found. No dependency between drug plasma levels and arrhythmia termination was found. E 047/1 has proven to be efficient and safe in the treatment of arrhythmia. E 047/1 is characterized by rapid plasma elimination, absence of proarrhythmic or cardiodepressive effects, mild adverse events, and excellent local tolerability. For further investigation, we recommend a combined bolus- and weight-adapted infusion regimen. PMID- 10813373 TI - Nicorandil metabolism in rat myocardial mitochondria. AB - The in vitro study using rats was carried out to clarify the hypothesis that nicorandil is denitrated and then may produce nitric oxide (NO) in myocardial mitochondria. In the presence of a NADPH-generating system, [14C]nicorandil, which was incubated in mitochondrial and microsomal fractions of the lung, heart, or liver, was converted to its main denitrated metabolite, SG-86 and other metabolites. Apparent Km and Vmax for nicorandil in mitochondrial and microsomal fractions of the heart were considerably similar to those of the lung, but completely different from those of the liver. It seems that glutathione-S transferases (GSTs) are not primarily involved in the conversion of nicorandil to SG-86, because a known GST inhibitor, indomethacin, did not affect the nicorandil degradation in the mitochondrial fraction. Nitrite, the stable metabolite of NO, was measured by the Griess reaction. In the presence of an NADPH-generating system, nicorandil significantly increased nitrite production in myocardial mitochondria, but SG-86 did not. These data strongly indicate that nicorandil is metabolized to SG-86 in myocardial mitochondria, then releasing NO, and that GSTs are not primarily responsible for the conversion of nicorandil to SG-86. PMID- 10813375 TI - Beneficial effects of fluvastatin on myocardial blood flow at two time-points in hypercholesterolemic patients with coronary artery disease. AB - Hypercholesterolemia is a major risk factor initiating and accelerating atherosclerosis and leading to severe stages of coronary artery disease (CAD) with a high risk of cardiovascular events. We investigated the impact of lipid lowering in patients with hypercholesterolemia and evident CAD on clinically relevant parameters like myocardial perfusion. Myocardial imaging was performed with thallium-201 single photon-emission computed tomography at rest and after maximal bicycle exercise in 22 patients after a 4-week lead-in period, and after 12 and 24 weeks of therapy with fluvastatin. Perfusion defects occurred in all patients, indicating stress-induced myocardial ischemia. After 12 weeks of therapy, the perfusion of the ischemic segments increased by 26% (277+/-99 to 349+/-96 cpm; p < 0.001), whereas the value of the normal segments was augmented only by 4% (478+/-44 to 497+/-28 cpm; p < 0.05). The results slightly improved further after 24 weeks. Moreover, a subgroup analysis elucidated a more pronounced effect in patients without lipid-lowering premedication. This nonpretreated group (n = 11) revealed an improvement of ischemic segments at stress by 42% at week 24. In contrast, pretreated patients had an increase of only 18% (between groups, p < 0.05), indicating a carryover effect of premedication. In conclusion, short-term therapy with fluvastatin acts beneficially on impaired vascular function in hypercholesterolemic patients with CAD. PMID- 10813374 TI - Vasoconstrictor effects of iso-prostaglandin F2alpha type-III (8-iso prostaglandin F2alpha) on human saphenous veins. AB - Free radical generation can initiate the peroxidation of arachidonic acid, resulting in a non-cyclooxygenase-dependent production of bioactive prostaglandin F2-like compounds. We have investigated the effects of iso-prostaglandin F2alpha type III, (iPF2alpha-III, formerly named 8-iso prostaglandin F2alpha) on human saphenous veins, and characterized the underlying mechanisms. In organ baths, the contractile effects of iPF2alpha-III were tested on saphenous vein rings coming from 22 patients. iPF2alpha-III induced concentration-dependent contractions of isolated human saphenous veins. The maximal contraction did not differ significantly from that of prostaglandin F2alpha (PGF2alpha). The pD2 values for iPF2alpha-III, PGF2alpha, endothelin-1 (ET-1), and U46619 (a stable thromboxane A2 mimetic) were 6.31+/-0.12, 5.66+/-0.13, 7.37+/-0.08, and 7.99+/-0.31, respectively (p < 0.001 for U46619 vs. iPF2alpha-III and PGF2alpha; and ET-1 vs. PGF2alpha). Emax values of iPF2alpha-III, PGF2alpha, ET-1, and U46619 were 137.7+/-24.3%, 145.9+/-7.5%, 92.9+/-16.8%, and 238.7+/-23.7%, respectively (p < 0.001 for U46619 vs. iPF2alpha-III, PGF2alpha and ET-1; and for PGF2alpha vs. ET 1). The responses to iPF2alpha-III were inhibited by GR 32191 10(-7) M, a TP receptor antagonist, without affecting the maximal response (pD2 values were 5.98+/-0.06 in the absence, and 5.22+/-0.05 in the presence of GR32191; p < 0.001). Concentration-effect curves to iPF2alpha-III were not affected by phosphoramidon 10(-5) M (an endothelin converting enzyme inhibitor), BQ123 10(-6) M (a selective ET(A)-receptor antagonist), BQ788 10(-6) M (a selective ET(B) receptor antagonist), and indomethacin 10(-5) M (a cyclooxygenase inhibitor). Finally, the contractile response of iPF2alpha-III did not involve the release of thromboxane B2 and ET-1, measured using enzyme immunoassays. This study demonstrates that iPF2alpha-III is a vasoconstrictor of human saphenous veins, with a potency fourfold greater than that of PGF2alpha, and 50 times less than that of the thromboxane A2 mimetic, U46619. These effects are mediated at least in part by TP-receptor stimulation, but do not involve thromboxane A2 or ET-1 release. PMID- 10813376 TI - Comparison of cardiovascular effects of a new calcium channel blocker AH-1058 with those of verapamil assessed in blood-perfused canine heart preparations. AB - The cardiovascular effects of AH-1058, a novel calcium channel blocker, were examined in comparison with those of verapamil using canine isolated, blood perfused papillary muscle, atrioventricular node, and sinoatrial node preparations. Intravenous administration of AH-1058 (20, 50, and 100 microg/kg) or verapamil (20, 50, and 100 microg/kg) to the blood-donor dog induced negative inotropic, dromotropic, and chronotropic effects and a coronary vasodilator action in cross-circulated isolated heart preparations, simultaneously inducing the same cardiac effects in the blood-donor dog. The order of potency of the effects of AH-1058 was ventricular contraction > coronary blood flow >> atrioventricular conduction > sinoatrial automaticity, whereas that of verapamil was coronary blood flow >> atrioventricular conduction >> sinoatrial automaticity > ventricular contraction. The cardiosuppressive effects of AH-1058, especially on ventricular contraction, were slower in onset and longer lasting than those of verapamil. These results suggest that this unique cardiovascular profile of AH 1058 may become beneficial for the treatment of certain pathologic processes, in which selective inhibition of ventricular calcium channels would be essential for drug therapy. PMID- 10813377 TI - Troglitazone inhibits mitogenic signaling by insulin in vascular smooth muscle cells. AB - Troglitazone (TRO) is an oral insulin-sensitizer that has direct effects on the vasculature to inhibit cell growth and migration. In vascular smooth muscle cells (VSMCs), insulin transduces a mitogenic signal that is dependent on the ERK1/2 MAP kinases. We examined the effects of TRO on this pathway and found that it inhibits mitogenic signaling. In quiescent VSMCs, insulin (1 microM) induced a 3.2-fold increase in DNA synthesis. TRO (1-20 microM) inhibited insulin stimulated DNA synthesis by 72.8% at the maximal concentration. TRO at I and 10 microM had no significant effect on insulin-stimulated ERK1/2 activity. At 20 microM, however, TRO modestly enhanced insulin-stimulated ERK1/2 activity by 1.5 fold. ERKs transduce a mitogenic signal by phosphorylating transcription factors such as Elk-1. which regulate critical growth-response genes. We used GAL-Elk-1 expression plasmids to detect ERK-dependent activation of Elk-1. TRO at 1-20 microM potently inhibited insulin-stimulated, ERK1/2-dependent Elk-1 transcription factor activity. Neither early steps in insulin signaling nor the phosphatidylinositol 3-kinase (PI3K) branch of this pathway were affected by TRO, because it had no effect on IRS-1 phosphorylation, PI3K/IRS-1 association, or Akt phosphorylation. Because TRO is a known ligand for the nuclear transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma), we tested two other ligands for this receptor, rosiglitazone (RSG) and 15-deoxy-delta12,14 prostaglandin J2 (15d-PGJ2). Both also inhibited insulin-induced DNA synthesis. In summary, these data show that TRO inhibits mitogenic signaling by insulin at a point distal of ERK1/2 activation, potentially by a PPARgamma-mediated inhibition of ERK-dependent phosphorylation and activation of nuclear transcription factors that regulate cell growth. PMID- 10813378 TI - Adenosine and angiotensin system interact in the regulation of renal microcirculation in humans. AB - We sought to evaluate the possible interaction between the adenosine and angiotensin systems in the regulation of renal microcirculation in humans. Twenty normotensive patients entered the study. Ten patients (group 1) were pretreated with 50 mg of captopril, an inhibitor of angiotensin-converting enzyme, whereas 10 patients (group 2) were pretreated with placebo. Incremental doses of adenosine (from 10(-5) to 1 mg) were injected into a renal artery to all patients at 5-min intervals. Adenosine injection reduced mean renal blood flow velocity in both groups (from 17.3+/-2.8 and 16.7+/-2 cm/s to 5.1+/-1.1 and 3.8+/-0.8 cm/s, in groups 1 and 2, respectively). The decrease in flow velocity was immediate after adenosine, and its duration was proportional to dosage (y = 3.05 x -2.7; R2 = 0.46; p < 0.01). However, group 1 had a slope of regression lower than group 2 (2.37 vs. 3.82 s; p < 0.03). The index of renal resistance (mean arterial pressure/mean blood flow velocity) increased linearly in both groups with adenosine, but group I showed a lower slope of increment (2.77 vs. 5.57 mm Hg/cm/s; p < 0.01). Adenosine administration induced a marked and transient increase in human renal resistance. This vasoconstrictive effect of adenosine was blunted by captopril pretreatment. PMID- 10813379 TI - Disparate effects of carvedilol versus metoprolol treatment of stroke-prone spontaneously hypertensive rats on endothelial function of resistance arteries. AB - In human hypertension, blockade of beta-adrenoceptors does not improve resistance artery structure or endothelial dysfunction. We tested in hypertensive rats the hypothesis that carvedilol, a beta-blocker with antioxidant properties, would improve endothelial dysfunction, whereas the beta1-selective blocker, metoprolol, would not. Twenty-week-old SHRSP were treated orally for 10 weeks with carvedilol (50 mg/kg/day) or metoprolol (100 mg/kg/day), with or without hydralazine (25 mg/kg/day), the latter because neither beta-blocker was a very effective blood pressure-lowering agent in this model. Mesenteric arteries (lumen, <300 microm) were studied on a pressurized myograph. After 10 weeks, untreated SHRSP had a systolic blood pressure (mm Hg) of 239+/-3 that was unaffected by carvedilol or metoprolol treatment but decreased (p < 0.05) by hydralazine (187+/-4), carvedilol + hydralazine (221+/-3), and metoprolol + hydralazine (197+/-3). Carvedilol alone improved endothelium-dependent relaxation of resistance arteries, as elicited by the lowest concentration of acetylcholine studied (10( 7) M), whereas metoprolol had no effect. Hydralazine improved endothelial function as elicited by acetylcholine at a dose of 10(-6) M, also found under cotreatment with carvedilol but attenuated by cotreatment with metoprolol. Carvedilol or metoprolol alone had no significant effect on endothelium independent relaxation produced by a nitric oxide donor (sodium nitroprusside). However, vessels from rats treated with carvedilol + hydralazine exhibited significantly greater relaxation than those from rats treated with metoprolol + hydralazine. These data suggest that carvedilol may have favorable effects on hypertension-related endothelial dysfunction not observed with metoprolol. Neither drug corrected small artery structure in SHRSP. PMID- 10813380 TI - Cardiac swelling-induced chloride current is enhanced by endothelin. AB - Endothelins (ETs) are a family of peptide hormones that act on G protein-coupled ET(A) and ET(B) receptors. ETs exert inotropic and chronotropic actions in the heart. Myocardial ischemia is associated with increased plasma levels of ET and cell swelling. We examined the effect of ETs on dog atrial swelling-induced chloride current (I(Cl,swell)). Whole-cell patch clamp was used; 10 nM ET-1 or ET 2 increased I(Cl,swell) by approximately twofold. ET-2 had no effect if I(Cl,swell) activation was prevented by hypertonic superfusate. Outward ET-2 induced current was blocked by 150 microM DIDS more effectively than inward current. Overnight pretreatment with phorbol 12-myristate 13-acetate (1.6 microM), pertussis toxin (100 ng/ml), or dialysis of the cell with 300 microM 2' deoxyadenosine 3'-monophosphate, a P-site inhibitor of adenylyl cyclase, did not diminish the effect of ET-2. The effect of ET-2 was blocked by an ET(A1)- (BQ123), but not an ET(B)-selective (BQ788) antagonist. ET-2-induced currents were inhibited approximately 70% by PD 98059 (30 microM), a selective MAPK kinase (MEK) blocker. PD 98059 did not affect basal whole cell current or I(Cl,swell) before exposure to ET-2. The data suggest that MEK activity is not required for activation of atrial I(Cl,swell) but that ET-2 stimulates I(Cl,swell) by a MEK dependent pathway. PMID- 10813381 TI - Effects of long-term treatment with nonselective endothelin receptor antagonist, TAK-044, on remodeling of cardiovascular system with sustained volume overload. AB - To assess the role of endothelin-1 (ET-1) on cardiovascular remodeling, nonselective endothelin-receptor antagonist TAK-044 was administered for the long term to rabbits with or without arteriovenous (A-V) shunt formation. Six weeks after sham operation (n = 12) or carotid-jugular shunt formation (n = 21), TAK 044 (30 mg/day) or saline was infused subcutaneously using osmotic mini pumps for another 6 weeks. Twelve weeks after operation, left ventricular (LV) diameter was enlarged with the presence of an A-V shunt; however, the levels of LV diameter and arterial pressure or the postmortem weight of LVs of shunt rabbits were similar between saline and TAK-044 groups. A linear relation of the luminal diameter and the medial cross-sectional area of the left and right carotid arteries was similar between shunt + saline and shunt + TAK-044 groups. In saline groups, myocardial ET-1 levels were higher in shunt than in sham rabbits (217+/ 22 vs. 136+/-19 pg/g tissue; p < 0.01 between rabbit groups) without changes in plasma ET-1 concentrations during saline infusion for 6 weeks. Differences in plasma ET-1 levels before and 6 weeks after the administration of TAK-044 were 0.32+/-0.78 and 0.16+/-0.28 pg/ml (NS between periods) in shunt and sham groups, respectively. In TAK-044 groups, myocardial ET-I levels 12 weeks after operation were similarly lower in both sham (105+/-7.4 pg/g tissue) and shunt rabbits (126+/-9.2 pg/g tissue) than in those with saline administration; however, the plasma ET-1 concentrations were increased significantly 6 weeks after TAK-044 administration by 5.0+/-0.6-fold and 3.5 +/-0.3-fold (p < 0.01) of the levels 6 weeks after operation in shunt and sham groups (NS between groups), respectively. Accordingly, myocardial but not plasma ET-1 levels were increased by a long-term burden of volume overload and were attenuated by a long-term administration of TAK-044 without altering drastically the hemodynamics or vascular remodeling. These results suggest that endogenous ET-1 does not play a major role in the compensatory stage of cardiovascular remodeling in the present volume-overload model. PMID- 10813382 TI - The negative inotropic effect of beta3-adrenoceptor stimulation in the beating guinea pig heart. AB - Although beta3-adrenoceptors (ARs) have been extensively characterized in brown and white adipocytes, their actions in the beating heart are unclear. We examined the effects of a beta3-AR agonist, BRL37344, on cardiac function and calcium transients in Langendorff-perfused guinea pig hearts by simultaneously measuring left ventricular (LV) pressure and Ca2+-dependent indo-1 fluorescence. BRL37344 induced a dose-dependent negative inotropic effect at concentrations from 10(-11) to 10(-8) M. Maximally, LV developed pressure decreased to 80+/-2%, +dP/dt to 81+/-2%. and -dP/dt to 81+/-3% of their respective control values (p < 0.01). The amplitude of the Ca2+ transient also decreased (to 92+/-3% of the control level; p < 0.01). The BRL37344 dose-response curve was not altered by nadolol (10(-5) M), a potent beta1- and beta2-AR antagonist, but completely suppressed by bupranolol (10(-6) M), a potent beta1-, beta2- and beta3-AR antagonist. To assess the potential role of a nitric oxide synthase (NOS) pathway, we determined whether the NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), modified the contractile response to BRL37344. L-NAME (10(-7) and 10(-4) M) attenuated the negative inotropic effects on LV developed pressure by 35 and 50%, suggesting that beta3-AR stimulation induces a negative inotropic effect on guinea pig hearts partly through a decrease in the Ca2+ transient and partly by the NOS pathway. PMID- 10813383 TI - Actions of L-NAME and methylene blue on the hypotensive effects of clonidine and rilmenidine in the anesthetized rat. AB - The antihypertensive mechanism of alpha2-adrenoceptor agonists, such as clonidine and rilmenidine, is not completely elucidated, although it is probably due to reduction of sympathetic tone mediated by stimulation of central alpha2 adrenoceptors. Because activation of alpha2-adrenoceptors on endothelial cells induces release of endothelium-derived relaxing factor (EDRF), we determined whether nitric oxide (NO) release is involved in the antihypertensive action of clonidine and rilmenidine. In chloralose-anesthetised Wistar rats, systolic and diastolic arterial blood pressures were recorded on a polygraph. Intravenous injection of clonidine or rilmenidine (control group) caused a rapid increase of arterial blood pressure. followed by a long-lasting hypotensive effect. The hypotensive effects, estimated as the area enclosed by the decrease in diastolic pressure during the 20 min after clonidine and rilmenidine injections, were 574+/ 60 and 410+/-59 mm Hg/min, respectively. The delta decrease in diastolic arterial blood pressure observed 20 min after intravenous injections of clonidine and rilmenidine was 48+/-5 and 34+/-3 mm Hg, respectively. Clonidine and rilmenidine injected 5-10 min after intravenous pretreatment with L-NAME (2 and 1 mg/kg) or methylene blue (10 mg/kg) induced hypotensive effects that were significantly smaller than that observed for the control group. These results suggest that the antihypertensive effects of clonidine and rilmenidine also may be modulated by the NO-cyclic guanosine monophosphate (cGMP) pathway at the level of the central nervous system and/or at the vascular peripheral circulation. PMID- 10813384 TI - Effects of ZK-807834, a novel inhibitor of factor Xa, on arterial and venous thrombosis in rabbits. AB - Inhibition of factor Xa (FXa) may interrupt thrombus progression. This study compared the antithrombotic activity of a novel FXa inhibitor, ZK-807834 [MW, 527 D; Ki (human FXa), 0.11 nM], with recombinant tick anticoagulant peptide [rTAP; MW, 6,685 D; Ki, (human FXa) = 0.28 nM], and DX-9065a [MW 445 D, Ki (human FXa), 40 nM] in rabbits with arterial thrombosis induced by electrical vascular injury. ZK-807834 also was compared with low molecular weight heparin (LMWH; MW, 5,500 D) during venous thrombosis induced by placing a copper wire and threads in the vena cava. Inhibitors were administered as an i.v. bolus and 2-h infusion. Total dosages of ZK-807834, > or =0.7 micromol/kg (n = 18); rTAP, > or =1 micromol/kg (n = 18); or DX-9065a, > or =11 micromol/kg (n = 18) decreased the incidence of arterial thrombotic occlusion compared with control animals (p < 0.05). However, five of six animals given the lowest effective dosage of rTAP and four of six animals given DX-9065a bled from a surgical incision >5 min, but only two of six animals given ZK-807834 bled >5 min. Venous clot weights were reduced compared with controls for dosages of ZK-807834 > or =0.007 micromol/kg (n = 36) or LMWH > or =0.2 micromol/kg (n = 18). Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were unchanged from baseline at the minimally effective dose of ZK-807834, whereas aPTT was increased twofold at the effective dose of LMWH. Thus ZK-807834 may be useful to attenuate thrombosis at lower dosages and with less perturbation of systemic hemostasis compared with available agents. PMID- 10813385 TI - Effects of prolonged propranolol treatment on left ventricular remodeling and oxidative stress after myocardial infarction in rats. AB - Mechanisms determining the benefit of beta blockade in patients with heart failure remain incompletely understood but are assumed consequent to prevention of deleterious effects of catecholamines. Recent studies have demonstrated that oxidative stress in congestive heart failure may be related to increased catecholamine levels. The aim of this study was to examine effects of long-term treatment with propranolol on progression of left ventricular (LV) dysfunction, remodeling and oxidative stress on an experimental model of chronic heart failure. Six weeks after myocardial infarction by coronary ligation, Wistar rats were randomized to two groups: 10 weeks of therapy with propranolol (50 mg/kg/day in drinking water) and no treatment (infarcted controls). A third group was sham operated rats without treatment. Animals were anesthetized for hemodynamic measurements, and hearts were then removed for histologic analysis, papillary muscle contractility study, and oxidative stress measurements using thiobarbituric acid reactive substance (TBARS) determination. Control infarcted rats demonstrated significant alterations of hemodynamic parameters and remodeling with increase of heart weight/body weight, of right ventricular lateral wall thickness, of LV circumference, LV septal area/body weight, and LV papillary muscle weight/body weight as compared with sham. In propranolol-treated rats, hypertrophy of the LV septum, papillary muscle, and right ventricle were similar to those of the infarcted control. Myocardial oxidative stress was significantly increased in control infarcted rats compared with sham, and propranolol prevented such oxidative stress increase. Papillary muscle isometric tension parameters were not significantly different among groups. Propranolol treatment prevented isoprenaline-induced spontaneous papillary muscle activity in vitro. Oxidative stress is increased in the rat model of heart failure secondary to coronary ligation. Long-term treatment with propranolol in vivo does not modify the compensatory process of hypertrophy but completely abolishes the oxidative stress increase and reduces the increased cardiac sensitivity to catecholamine-induced arrhythmias observed in this experimental model of heart failure. PMID- 10813386 TI - Agonist-dependent difference in the mechanisms involved in Ca2+ sensitization of smooth muscle of porcine coronary artery. AB - This study was undertaken to explore possible signal-transduction mechanisms involved in the Ca2+-sensitizing effects of carbachol and endothelin-1 (ET-1) by using beta-escin-skinned smooth muscle of porcine coronary artery. Pretreatment with C3 exoenzyme of Clostridium botulinum, which selectively inactivates rho p21 by adenosine diphosphate (ADP) ribosylation, resulted in a significant inhibition of ET-1-induced Ca2+ sensitization, but had no effect on carbachol-induced Ca2+ sensitization. Whereas the protein kinase C (PKC) inhibitors calphostin C and staurosporine did not affect the Ca2+-sensitizing effect of carbachol, the tyrosine kinase inhibitors genistein and tyrphostin 25 greatly but incompletely suppressed it. In contrast, the Ca2+-sensitizing effect of ET-1 was significantly inhibited by either calphostin C or genistein. Although the inhibitory effect of calphostin C on ET-1-induced Ca2+ sensitization was less than that of genistein, the effects of calphostin C and genistein were additive. The genistein-sensitive component of ET-1-induced Ca2+ sensitization appeared to include the C3-sensitive one. However, a substantial enhancement by ET-1 of the Ca2+-induced contraction was observed even in the presence of the two inhibitors. In beta-escin-skinned smooth muscle of rabbit mesenteric artery, ET-1-induced Ca2+ sensitization was marginally affected by C3 pretreatment, calphostin C, and genistein. We conclude that, although PKC activation and rho p21 protein-dependent and -independent tyrosine phosphorylation each plays an important role in an increase in myofilament Ca2+ sensitivity, the contributions of these signaling pathways to Ca2+ sensitization are different depending on receptor agonists and tissues used. Furthermore, these data suggest the existence of an as yet undefined signal transduction mechanism involved in Ca2+ sensitization caused by receptor agonists. PMID- 10813387 TI - Increased expression of L-type calcium channels in vascular smooth muscle cells at spastic site in a porcine model of coronary artery spasm. AB - Coronary artery spasm is caused primarily by increased contractility of vascular smooth muscle. Excessive Ca2+ entry into vascular smooth muscle cells (VSMCs) may be one of the key mechanisms for the spasm, but no study has ever directly examined the possible alterations of Ca2+ channels in the spastic coronary artery. Here we show that L-type Ca2+ channels are excessively expressed at the spastic site of the coronary artery. In a porcine model of coronary spasm with balloon injury, both receptor-mediated stimulation of L-type Ca2+ channels by serotonin and direct stimulation of the channels by Bay K 8644 (a dihydropyridine Ca2+ channel agonist) repeatedly induced coronary spasm in vivo, which was abolished by pretreatment with nifedipine, a dihydropyridine Ca2+ channel antagonist. In a single VSMC freshly dispersed from coronary arteries in vitro, patch-clamp experiments showed that current density of L-type Ca2+ channel current was significantly increased in VSMCs from the spastic site compared with that from the control site even when the channels were maximally stimulated by Bay K 8644. There was no difference in the sensitivity of the channels to Bay K 8644. These results indicate that functionally available L-type Ca2+ channels are excessively expressed at the spastic site of the coronary artery in our porcine model, suggesting that increased expression of L-type Ca2+ channels and concomitant increase in Ca2+ entry into VSMCs through the channels may contribute, at least in part, to the pathogenesis of coronary artery spasm. PMID- 10813388 TI - 25 years of contributions to characterizing gene expression and replication within the three-dimensional context of nuclear architecture. PMID- 10813389 TI - Understanding "active" chromatin: a historical perspective of chromatin remodeling. AB - Two phenomena have long been observed to correlate with transcriptionally active chromatin: increased histone acetylation and increased sensitivity to nucleases, including specific patterns of nuclease hypersensitivity in the promoters of active or inducible genes. Work in recent years has at last identified protein complexes required to form these hallmarks of active chromatin: histone acetyltransferases (HATs) and ATP-dependent chromatin remodeling complexes. This review traces the history of these discoveries, including the development of essential tools that allowed the major advances in the field, and describes the current understanding of the interactions between HATs and ATP-dependent remodelers. PMID- 10813391 TI - Specialized transcription factories within mammalian nuclei. AB - Recent evidence suggests that active RNA polymerases are concentrated in discrete 'factories' where they work together on many different templates. The evidence that such factories specialize in the transcription of particular groups of genes is reviewed. PMID- 10813390 TI - Cell nucleus in context. AB - The molecular pathways that participate in regulation of gene expression are being progressively unraveled. Extracellular signals, including the binding of extracellular matrix and soluble molecules to cell membrane receptors, activate specific signal transducers that process information inside the cell leading to alteration in gene expression. Some of these transducers when translocated to the cell nucleus may bind to transcription complexes and thereby modify the transcriptional activity of specific genes. However, the basic molecules involved in the regulation of gene expression are found in many different cell and tissue types; thus, the mechanisms underlying tissue-specific gene expression are still obscure. In this review we focus on the study of signals that are conveyed to the nucleus. We propose that the way in which extracellular signals are integrated may account for tissue-specific gene expression. We argue that the integration of signals depends on the nature of the structural organization of cells (i.e., extracellular matrix, membrane proteins, cytoskeleton, nucleus) that defines a particular cell type within a tissue. Thus, gene expression can be envisioned as being regulated by the mutual influence of extracellular and intracellular organizations, i.e., in context. PMID- 10813392 TI - Specification of chromatin domains and regulation of replication and transcription during development. AB - It is becoming increasingly clear that transcription control is carried out at several interconnecting levels. Besides nucleosomal organization and interaction between transcription factors and gene promoters and other regulatory elements, long-range organization of chromatin in loops or domains seems to play a role in transcriptional regulation. A similar organization is likely to be crucial in the control of the timing and selection of origins of DNA replication. This review considers the implications of domain organization of eukaryotic genome in developmental control of transcription and replication. PMID- 10813393 TI - Nuclear matrix targeting of steroid receptors: specific signal sequences and acceptor proteins. AB - The nuclear matrix provides the framework for various processes that occur within the nucleus such as transcription, replication, and splicing. As the composition of the nuclear matrix varies between different cell types, the matrix may influence cell-type-specific gene expression. A number of transcription factors have been shown to be associated with the nuclear matrix, including steroid hormone receptors that were the first transcriptional regulatory proteins localized to this compartment. In this review we highlight the most recent advances in our understanding of steroid hormone receptor targeting to the matrix. A specific nuclear matrix targeting signal (NMTS) has been identified within the glucocorticoid receptor (GR) that includes portions of its DNA-binding domain and tau2 transactivation domains. Distinct nuclear matrix acceptor proteins appear to interact within the GR NMTS and impart divergent effects on the transcriptional regulatory properties of the receptor. PMID- 10813394 TI - The nuclear matrix as a target for viral and cellular oncogenes. AB - As the key integrator of nuclear structure and function, the nuclear matrix is likely to be an important target for structural and functional alterations during the process of neoplastic transformation. Here I summarize and discuss data demonstrating that the major transforming protein of the small DNA tumor virus simian virus 40 (SV40), the SV40 large tumor antigen (large T), specifically targets the chromatin and the nuclear matrix during viral transformation. I then turn to recent evidence endorsing the concept that mutant p53--the most commonly expressed oncogene in human cancer--might exert its oncogenic activities by specifically interacting with the nuclear matrix. The data suggest that SV40 large T and mutant p53 might be members of a new family of oncogenes that exert their oncogenic functions by directly modulating nuclear structure and function. PMID- 10813395 TI - Caught in the act: binding of Ku and PARP to MARs reveals novel aspects of their functional interaction. AB - Specific regions of eukaryotic genomic DNA that exhibit high-affinity binding to the nuclear matrix in vitro are called matrix attachment regions (MARs) and are implicated in the loop domain organization of chromatin. Small regions possessing high base unpairing potential within these MARs are referred to as base unpairing regions (BURs). BUR-affinity chromatographic separations of proteins from breast cancer cells yielded, almost exclusively, a mixture of poly (ADP-ribose) polymerase (PARP) and DNA-dependent protein kinase (DNA-PK), two nuclear enzymes that are implicated in the cellular response to DNA damage. Contrary to the long held notion that PARP and Ku autoantigen, the DNA-binding heterodimeric subunit of DNA-PK, bind only to DNA ends, recently we have shown that both proteins individually bind BURs with high affinity and specificity in an end-independent manner. Furthermore, Ku autoantigen forms a molecular complex with PARP in the absence of DNA, and the physical association of these proteins synergistically enhanced their BUR-binding activity. Autoribosylation of PARP abolished its association with Ku autoantigen and BUR-binding activity. These findings have, for the first time, provided a molecular link toward elucidating the functional interaction between PARP and DNA-PK. The identification of MARs as their common binding target suggests a novel role for these enzymes in the modulation of chromatin structure and function. PMID- 10813396 TI - Transcriptional augmentation: modulation of gene expression by scaffold/matrix attached regions (S/MAR elements). AB - For a long time S/MARs could only be characterized by the assays in vitro that led to their detection. Only recently a number of biological activities emerged that are common to most or all S/MARs that are detected by the classic procedures. This review focuses on the phenomenon of transcriptional augmentation that is found for genomically anchored or episomal genes and on a group of partially overlapping activities that are suited to maintain an episomal status. Further, it is attempted to correlate properties of the S/MAR-scaffold interaction with prominent or prototype protein binding partners. PMID- 10813397 TI - On the relationship of matrix association and DNA replication. AB - The nuclear matrix is believed to contain sites of assembly of protein complexes that catalyze the initiation of DNA replication as well as DNA elongation. To explore this relationship, DNA replicated by human fibroblasts at the beginning of the S phase was purified and used to construct a cosmid library. Hybridization studies with a subgroup of clones (about one-sixth of the total clones in this library) showed that many of them were highlighted by probes prepared from early replicating DNA, as well as from nuclear matrix-associated DNA. Statistical analysis showed a positive correlation between these hybridization results. We seek to identify origins of replication that are activated early in the S phase of the cell cycle in human cells. Therefore, clones isolated from this library are being analyzed for the presence of structural motifs that have been found in other origins of replication and for potential sites of attachment to the nuclear matrix. This method of analysis is illustrated here using the published sequences for the origins of replication reported for the human lamin B2 and HPRT genes. PMID- 10813398 TI - The nuclear pore complex: structure, function, and dynamics. AB - A full understanding of nucleocytoplasmic transport depends on knowledge of nuclear pore complex (NPC) structure, the functional roles of NPC components, their interactions during transport and dynamics during the cell cycle. NPC structure is conserved, flexible, and is not simply a tunnel between the nucleus and cytoplasm but appears to be actively involved in the transport process by a series of structural modifications. Transport through the NPC begins in either of its asymmetrical peripheral compartments that are both structurally reorganized during transport in different ways. The central compartment is composed of two symmetrical halves, and functions as a system of transiently open, discrete gates that is not believed to play a role in determining direction. Each NPC subunit has a specific morphology that corresponds to the functional role it plays. A complicated system of vertical and horizontal connections may allow one part of the NPC to transmit a signal to other parts, leading to an ordered series of conformational changes that drive translocation. High-resolution scanning electron microscopy has identified sequential stages of NPC assembly in vitro and revealed how the individual NPC components are assembled into a mature NPC. This review focuses on structural events during transport and on possible mechanisms of NPC assembly. PMID- 10813433 TI - Rewarming from hypothermia leads to elevated plasma lipopolysaccharide concentrations. AB - Rewarming victims of hypothermia such as divers or immersion victims, participants in winter sports and military operations, and surgical patients on cardiopulmonary bypass (CPB) may lead to vascular instability, multiorgan failure, shock, and even death. While the causes of these rewarming symptoms are unknown, they may be related to bacterial lipopolysaccharide (LPS) translocated from the intestines into the circulation due to splanchnic ischemia. We have determined LPS during the cooling (to 31.5 degrees-34.0 degrees C) and rewarming phases of hypothermic surgery in 11 patients at the Stanford University Medical Center. During rewarming, there was an LPS spike in 6/11, in one more patient there was an LPS spike during surgery but not during rewarming, and in 4/11 there was no rise in LPS, i.e., a temporary endotoxemia occurred in 7/11 (63.6%) patients, usually at the commencement of rewarming. All four patients with no LPS spike received dexamethasone for at least 7 days before surgery. We propose that hypothermia reduced splanchnic blood flow (BF), causing ischemic damage to the gut wall and translocation of LPS from the gut into the vascular space. Upon rewarming, splanchnic BF is restored, the translocated LPS transits from the splanchnic to the systemic circulations as a bolus, and the gut wall is healed. No sequelae occurred in these patients because of their adequately functioning immune systems. However, had they been immunocompromised, symptoms might have occurred. Rewarming of accident victims probably also incurs a similar risk of endotoxemia, and dexamethasone may have protected the gut wall. Further studies are indicated. PMID- 10813434 TI - Effects of weather on incidence of attempted suicide by carbon monoxide poisoning. AB - Depressive illness and related suicide attempts have been reported to have seasonal variation, possibly related to weather. This study sought to determine the effects of weather and time of year on suicide attempts by carbon monoxide (CO) poisoning. Cases of patients in western Washington State attempting suicide by CO poisoning and referred to a regional hyperbaric oxygen treatment center from 1981-1995 were reviewed retrospectively. Information from the National Climatic Data Center was used to compare date of treatment to local weather data on the day of attempted suicide and on each day of the preceding week. The study population consisted of 264 patients attempting suicide by CO poisoning on 251 days of the 15-yr period. Mean and total precipitation in the preceding 7 days strongly correlated with attempted suicide rate, with incident rate ratios ranging from 1.75 to 2.77 and 1.14 to 1.75, respectively (P values <0.05). A decrease in minimum daily temperature also correlated with attempted suicide rate. No other weather variables showed significant correlation with dates of CO suicide attempts. February, March, and October were seen to be separate factors related to a higher incidence of attempted suicide by CO poisoning. Precipitation in Seattle, change in minimum temperature, and month of the year correlated with the regional incidence of suicide attempts with CO poisoning. PMID- 10813436 TI - Screening for oxygen sensitivity in U.S. Navy combat swimmers. AB - The United States Naval Special Warfare Community uses oxygen tolerance testing to screen Navy combat swimmer candidates for sensitivity to the toxic effects of hyperbaric oxygen (HBO2). Between 1976 and 1997, 6,250 oxygen-tolerance tests were performed at the Naval Special Warfare Center and Naval Special Warfare Group One. Our review of these data found only six episodes of O2 toxicity for an incidence of 0.096%, an incidence considerably lower than the rate of 1.9% reported in an earlier review using data from the Naval Safety Center. Additionally, we reviewed data from the Naval Safety Center from 1986 to 1997 and found only one episode of O2 toxicity among 157,930 LAR V dives. Many factors other than individual sensitivity to HBO2 may contribute to the occurrence of O2 toxicity episodes during combat swimmer operations. The authors conclude that O2 tolerance testing of U. S. Navy SEAL candidates is not a useful screening test and recommend discontinuation of this test. PMID- 10813435 TI - Hyperbaric oxygen in the treatment of delayed radiation injuries of the extremities. AB - Hyperbaric oxygen (HBO2) is used as an adjunct in the treatment of radiation injury at many sites, including the mandible, larynx, chest wall, bladder, and rectum. In these disorders, HBO2 is effective in stimulating neovascularization and reducing fibrosis. No previous publications report the application of HBO2 to radiation injuries of the extremities. From 1979 until 1997, 17 patients were treated at the Southwest Texas Methodist and Nix Hospitals for nonhealing necrotic wounds of the extremities within previously irradiated fields. All but one wound involved a lower extremity. Most of the patients had been irradiated for soft tissue sarcomas or skin cancers. The rest were irradiated for a variety of malignancies. HBO2 was delivered in a multiplace chamber at 2.4 atm abs daily for 90 min of 100% oxygen at pressure. This report is a retrospective, uncontrolled review of these patients. Eleven patients (65%) healed completely whereas five (29%) failed to heal and one (6%) was lost to follow-up. Three (60%) of those who failed were found to have local or distant recurrence of their tumor early in their course of hyperbaric treatment and were discontinued from therapy at that time. When last seen in the clinic, the wound of the patient who was lost to follow-up was improved but not completely healed. Four of those who failed (including the two with local tumor recurrence) required amputation. If we exclude those with active cancer and the patient lost to follow-up, the success rate was 11 of 13 or 85%. HBO2 was applied successfully with complete wound healing and the avoidance of amputation in a majority of these patients. The consequences of failure in patients suffering from radiation necrosis of the extremities (some complicated by the presence of tumor) are significant, with 80% of the five failures requiring amputation. In radiation injuries of the extremities as in delayed radiation injury at other sites, HBO2 is a useful adjunct and should be part of the overall management. PMID- 10813437 TI - Lack of effect of anti-C5a monoclonal antibody on endothelial injury by gas bubbles in the rabbit after decompression. AB - Previous studies have shown that gas bubbles activate the complement system in vitro, generating C5a. The effect of anti-C5a monoclonal antibody 4B1C11 in preventing endothelial damage caused by decompression in the pulmonary artery of the rabbit was examined. The endothelial response was measured using tension measurements in the blood vessel wall. The mean bubble count for all rabbits (n = 24) was 4.2+/-3.1 bubbles x cm(-2), and ranged from 0 to 15 bubbles x cm(-2). Animals with many bubbles showed significantly more vascular damage than those with fewer bubbles. Anti-C5a monoclonal antibody could not prevent endothelial damage than that occurred after exposure to this level of gas bubbles. The maximum number of gas bubbles present is important for the endothelial damage. We speculate that the endothelial damage observed was mainly mechanical. A possible beneficial effect of anti-C5a antibody can thus be masked at a high degree of bubble generation. This study, together with a previous paper, demonstrates that gas bubbles cause endothelial damage from decompression both in the pig and in the rabbit. PMID- 10813438 TI - Hyperbaric stress during saturation diving induces lymphocyte subset changes and heat shock protein expression. AB - To clarify the cellular responses and biochemical markers of hyperbaric stress, we investigated heat shock protein (hsp) expression and subset changes of human peripheral blood lymphocytes during saturation diving. Five healthy male subjects underwent a 39-day saturation dive to the maximal storage pressure of 4.1 MPa [400 meters of sea water (msw)]. During the saturation dive, lymphocyte subset changes were detected using a flow cytometer, and increased expressions of hsp 72/73 and hsp 27 were observed by Western blot analysis. Lymphocyte subset changes included a decrease in CD4:CD8 ratio and in the fraction of CD4+ T cells as well as an increase in NK cells, especially during the 400-msw bottom phase. An increased expression of hsp 27 compared to hsp 72/73 was obvious, especially during the hold period at 100 msw. These results suggest that changes in lymphocyte subsets and hsp expression are useful markers for stress responses during saturation diving. These changes may also be useful for testing the barotolerance of divers for saturation diving. PMID- 10813439 TI - Cigarette smoking and transcutaneous oxygen tensions: a case report. AB - We report the effects of acute smoking cessation on transcutaneous oxygen (PtcO2) measurements in room air and with hyperbaric oxygen (HBO2) of an extremity at risk for amputation. The reports on cigarette smoking and PtcO2 do not discuss acute smoking cessation. PtcO2 measured 46 h after smoking cessation increased 10% while breathing room air and 34% with HBO2, as compared to measurements made before smoking cessation. PMID- 10813440 TI - Hyperbaric oxygen vs. normobaric oxygen in carbon monoxide intoxication. PMID- 10813441 TI - Failure to assess motivation, need to consider psychiatric variables, and absence of comprehensive examination: a skeptical review of neuropsychologic assessment in carbon monoxide research. PMID- 10813442 TI - Strategic environmental management: time for a new approach. AB - Attempting to manage the environment as a whole, even from just a scientific standpoint, is not easy. The sheer size and scale can be daunting. It is difficult even to encompass the many facets involved. This short paper therefore attempts to tackle the problem by describing a framework approach which encompasses how one might assess the actual state of the environment at any one time; characterise the ever-changing pressures placed upon it; the manner by which the resultant problems need to be addressed; and thus what sort of responses are required. No system or framework is ever likely to be fully adequate, but some form of framework is essential in order to deliver a more sustainable approach to environmental management than has so far been achieved. PMID- 10813443 TI - Trends in risk assessment and risk management. AB - Environmental risk assessment has matured into a powerful analytical tool, which is finding ever-wider applications in the arena of policy making and regulation. However, the principal focus of its development to date has been on the technical challenges of characterising and modelling the environmental behaviour and biological action of chemicals, whereas issues concerning its broader socio political context have been generally neglected. Problem definition, risk analysis and decision making have, therefore, tended to be dominated by experts and by expert opinion. Fresh insights from the social sciences advocate a pluralistic, inclusive approach, with experts participating alongside other stakeholders in a consensual decision making process. Adoption of this paradigm has far reaching consequences for the form and conduct of risk assessment and risk management. PMID- 10813444 TI - Establishing the true costs and benefits of environmental protection and enhancement in the aquatic environment AB - In the aquatic environment, a number of techniques are available to evaluate environmental benefits arising from improvements in water quality. However, such techniques do not account for the environmental costs of implementing a given quality change, for example those associated with the construction and operation of necessary treatment processes. Life cycle assessment is a technique that enables the nature of environmental impacts arising from the construction and operation of processes to be established. Although comprehensive assessment of all options is likely to be uneconomic, focusing on a number of key system inputs and outputs offers the possibility to include such impacts cost effectively in investment appraisal. PMID- 10813445 TI - Landscapes, tourism, and conservation AB - One key aspect of global change is a decrease in ecological integrity as more and more landscapes are developed, leaving a mosaic of intact refuges and degraded patches that may not be sufficient for conserving biodiversity. While increases in human population and shifts in the distribution of people affect land use, the temporary movement of people can have major implications for conservation and biodiversity. Three examples are presented where recreation/tourism can enhance the conservation of land on a landscape scale, leading to habitat protection and biodiversity preservation: (1) Shorebirds often require a matrix of different habitat types during migratory stopovers, and ecotourism can serve as a catalyst for landscape scale protection of habitat. (2) Riparian habitats can serve as corridors to link diverse habitat patches, as well as serving as biodiversity hotspots. (3) Remediation and rehabilitation of contaminated lands, such as those of the US Department of Energy, aimed at developing recreational activities on the uncontaminated portions, can be the most economical form of re-development with no increase in human or ecological risk. Since large areas on many DOE sites have been undisturbed since the Second World War, when they were acquired, they contain unique or valuable ecosystems that serve an important role within their regional landscapes. In all three cases the judicious development of recreational/tourist interests can encourage both the conservation of habitats and the wise management of habitats on a landscape scale. While some species or habitats are too fragile for sustained tourism, many can be managed so that species, ecosystems and ecotourists flourish. By contributing to the economic base of regions, ecotourists/recreationists can influence the protection of land and biodiversity on a landscape scale, contributing to ecosystem management. The human dimensions of land preservation and biodiversity protection are key to long term sustainability, and ecotourists/recreationists can be one management option. PMID- 10813446 TI - The critical loads concept: milestone or millstone for the new millennium? AB - The significance of the introduction of the critical loads concept in the closing decades of the 20th century is considered critically, both in the context of its potential in planning pollution abatement strategies over a range of regional spatial and temporal scales and in terms of its more general impact upon atmospheric pollution effects research. Based upon perceived strengths and shortcomings of the concept and of the ways in which it has thus far been applied, a brief assessment is made also of its possible role in the opening decade of the new millennium. It is concluded that the validity of some of the fundamental underpinning concepts and of the data being used for their application could, and should, be questioned. Nevertheless, environmental pollution management and abatement policy planning will continue to require application of the critical loads concept or something very similar, both in existing and novel areas of application, for the sustainable management of soil and water resources in the long term. PMID- 10813447 TI - Compartmentalization in environmental science and the perversion of multiple thresholds. AB - Nature and living organisms are separated into compartments. The self-assembly of phospholipid micelles was as fundamental to the emergence of life and evolution as the formation of DNA precursors and their self-replication. Also, modern science owes much of its success to the study of single compartments, the dissection of complex structures and event chains into smaller study objects which can be manipulated with a set of more and more sophisticated equipment. However, in environmental science, these insights are obtained at a price: firstly, it is difficult to recognize, let alone to take into account what is lost during fragmentation and dissection; and secondly, artificial compartments such as scientific disciplines become self-sustaining, leading to new and unnecessary boundaries, subtly framing scientific culture and impeding progress in holistic understanding. The long-standing but fruitless quest to define dose effect relationships and thresholds for single toxic agents in our environment is a central part of the problem. Debating single-agent toxicity in splendid isolation is deeply flawed in view of a modern world where people are exposed to low levels of a multitude of genotoxic and non-genotoxic agents. Its potential danger lies in the unwarranted postulation of separate thresholds for agents with similar action. A unifying concept involving toxicology and radiation biology is needed for a full mechanistic assessment of environmental health risks. The threat of synergism may be less than expected, but this may also hold for the safety margin commonly thought to be a consequence of linear no-threshold dose effect relationship assumptions. PMID- 10813448 TI - Health-effects related structure-toxicity relationships: a paradigm for the first decade of the new millennium. AB - Prediction of the effects of industrial chemicals to humans will be an area of increasing concern in the next century. The role of quantitative structure activity relationships (QSARs) is to aid in the prediction of effects by determination of the limits of variation in structure that are consistent with the production of a specific toxic effect and define the ways in which alterations in structure influences toxicity. The paradigm followed in the development of QSARs for ecotoxic-endpoints has been successful as a direct result of the availability of in vivo toxicity data in which to build initial QSARs and validate surrogate test systems. However, the lack of quantitative in vivo toxicity data means this paradigm cannot be used in the prediction of human health-effect endpoints. Therefore, a new paradigm, which provides guidance in the use of predictive QSARs for health-effects that serves to circumvent the problems associated with the lack of whole-mammal toxicity data must be established. A scenario is given that provides for the development, standardization, and validation of health-effects related QSARs in the first decade of the 21st century. Due to the structural diversity and sheer number of industrial organic chemicals, assays used to garner health-effects data must be based on quantifiable, rapid, reliable, and inexpensive surrogate endpoints. New 'biotools' developed using modern molecular techniques will aid in the circumvention of in vivo health-effects data and provide measurable endpoints, which can be used as surrogates for health-effects endpoints. This has particular application in receptor-mediated toxicity and gene expression. The lack of whole mammal health-effects data means that the standardization and validation, of these endpoints will be accomplished in novel ways. Databases garnered using modern biotools will allow the derivation of QSARs developed for validated surrogate health-effect endpoints. If QSARs are to be useful in bridging gaps in health-effects data, they must be based on accurate, reproducible data for a robust series of non-congeneric chemicals. The latter applies to both toxicity and descriptor values. Because health-effects are often the result of metabolic activation, if these new QSARs are to be meaningful, validated software for predicting metabolite formation must be incorporated. Lastly, once QSARs and software are developed and validated, they will need to be linked into some type of expert system. PMID- 10813449 TI - Particulate matter in the atmosphere: which particle properties are important for its effects on health? AB - Whilst epidemiological studies have consistently demonstrated adverse effects of particulate matter exposure on human health, the mechanism of effect is currently unclear. One of the major issues is whether the toxicity of the particles resides in some particular fraction of the particles as defined by chemical composition or size. This article reviews selected data on the major and minor component composition of PM2.5 and PM10 particulate matter showing quite major geographic variations in composition which are not reflected in the exposure-response coefficients determined from the epidemiology which show remarkably little spatial variation. The issue of particle size is more difficult to address due to the scarcity of data. Overall, the data presented provides little support for the idea that any single major or trace component of the particulate matter is responsible for the adverse effects. The issue of particle size is currently unclear and more research is warranted. PMID- 10813450 TI - Links between environment and health: possible future directions. AB - Linkages between environmental quality and human health are likely to continue to be of major interest, but quantitatively smaller risks impacting on larger population groups will probably become a prime future focus. Increasingly the aim will be to understand the inter-relatedness of impacts on human health and to sustain human physical and mental well-being rather than simply preventing disease. PMID- 10813451 TI - The 'mysterious' disease in Swedish moose. Concentrations of trace elements in liver and kidneys and clinical chemistry. Comparison with experimental molybdenosis and copper deficiency in the goat. AB - Moose (Alces alces L.) affected by a disease with unknown aetiology were compared with healthy moose regarding trace element concentrations in liver and kidneys, as well as certain clinical chemical parameters of blood plasma. The diseased moose showed decreased hepatic concentrations of Cu, Cd and Mg. Renal concentrations of Cd, Co, Mg and Mn were also decreased. Substantially increased concentrations in both liver and kidneys were seen for Al, Ca, Fe, Pb and Zn, while Se and Mo were increased in liver tissue. The hepatic levels of Mo in the affected moose were 36% higher and Cu was approximately 60% lower than in reference animals collected in 1982. The most important clinical chemical changes were decreased concentrations of bilirubin, thyroxine (T4) and the liver-specific enzymes GLDH and g-GT. Also, the activities of the enzymes CuZn-SOD and GSH-Px in erythrocytes were decreased. Increases were recorded for free fatty acids (NEFA), the muscle-specific enzyme CK, but especially for urea and insulin. Changes in the plasma protein pattern were also discernible after electrophoresis on agarose gel membranes, indicating chronic immunostimulation. All the observed changes in trace element concentrations and clinical chemical parameters are compatible with molybdenosis and secondary Cu deficiency in ruminants and most of the parameters are in agreement with those found in experimental copper deficiency and molybdenosis in the goat. It is, therefore, suggested that the moose disease with previously unknown aetiology is mainly caused by molybdenosis. PMID- 10813453 TI - Experimental copper and chromium deficiency and additional molybdenum supplementation in goats. I. Feed consumption and weight development. AB - Secondary Cu deficiency, Cr deficiency and molybdenosis were suggested causes of the 'mysterious' disease afflicting moose (Alces alces L.) in a region in south west Sweden affected by acid rain. A model experiment with goats was performed to study the clinical chemical parameters, determine the tissue contents of trace and minor elements, to perform pathological and histopathological investigations and to compare the findings with those in moose disease. Twenty 3-month-old male goats were assigned to four dietary treatments (five animals each) in an experiment lasting for 20 months. The four groups in the study were: control group, Cu-deficient group (group 1), Cr-deficient group (group 2), and Cu- and Cr deficient group (group 3). The animals were fed a basic semi-synthetic diet. At the end of the study the three surviving animals of group 3 were supplemented with additional tetrathiomolybdate (TTM) during the last 2 months. Feed consumption and weight development of the animals were monitored and are presented. The feed consumption of the two Cu-deficient groups of goats (group 1 and group 3) supported the previously described observations in copper deficiency in ruminants, e.g. decreased appetite and feed intake. A previously unreported effect of Cr deficiency in ruminants is now described in goats. Chromium deficiency at adequate Cu supplementation (group 2), caused increased lipid synthesis and a weight gain of 32 kg compared with that of the control group (20 kg). A possible explanation for this unexpected weight increase in only Cr deficiency is discussed. It is concluded that the feeding experiment does not support the hypothesis concerning the relation of Cr deficiency to the moose disease. PMID- 10813452 TI - A syndrome of molybdenosis, copper deficiency, and type 2 diabetes in the moose population of south-west Sweden. AB - Since the mid-1980s, a 'mysterious' disease has been afflicting the moose (Alces alces L.) population of south-western Sweden. Molybdenosis combined with secondary copper deficiency syndrome has been suggested as the cause of the clinical signs and of necropsy findings, supported by trace element analysis. Copper deficiency has long been associated with disturbed carbohydrate metabolism and also with oxidative stress. When testing the oxidative stress hypothesis, we found increased concentrations of the glycoxidation products pentosidine and carboxymethyl-lysine (CML), both in plasma proteins and in renal tissue, when compared with control values. The concentration of glycated lysine (furosine), a marker of hyperglycaemia, was also increased. These data, together with elevated insulin levels in affected moose, strongly suggest that they are suffering from an environmentally-induced, non-insulin-dependent type 2 diabetes. PMID- 10813454 TI - Experimental copper and chromium deficiency and additional molybdenum supplementation in goats. II. Concentrations of trace and minor elements in liver, kidneys and ribs: haematology and clinical chemistry. AB - Since the mid-1980s a previously undescribed disease has affected moose in south western Sweden. Investigations made to reveal evidence of a viral aetiology have proved unsuccessful. Trace element studies in apparently healthy moose shot during regular hunting suggested a trace element imbalance, with excessive molybdenum uptake causing secondary copper deficiency. The results also indicated a possible chromium deficiency. To verify this hypothesis, an experimental study was performed in male goats fed a semi-synthetic diet for 1.5 years. The animals were kept and treated in four groups: Controls, Cu-deficient group (group 1), Cr deficient group (group 2), and combined Cu- and Cr-deficient group with additional supplementation of tetrathiomolybdate for 10 weeks at the end of the study (group 3). The present paper presents tissue contents of trace and minor elements, haematology and clinical chemical parameters. Feed consumption and weight development, as well as pathological and histopathological investigations, were also performed in this study, but these results are presented elsewhere. Changes in trace element concentrations were determined by comparing groups 1, 2 and 3 with the control group. Increased concentrations were observed for Al, Ca, Co, Fe, Mo, Pb, Se in the liver; for Al, Cd, Co, Cr, Mo in the kidneys; and for Mn and Mo in the ribs. Considerable accumulation of Mn in ribs seems to be a useful way to determine oxidative stress. Decreases in Mg and P in all organs and blood serum is characteristic of Cu deficiency and molybdenosis. Also the ratio of Ca/Mg was increased as the result of tissue lesions causing an intracellular increase in Ca and decrease in Mg. The trace element changes observed in group 1 were enhanced by the Mo supplementation in group 3, resulting in characteristic patterns, 'spectra' of changes. The alterations were not as remarkable in group 2 as in the two other groups. However, Cr deficiency considerably influenced Al, Co, V and to a smaller extent also Mn in ribs. In groups 1 and 2, only a few minor changes were detected in the haematological parameters, probably caused by increased adrenal activity after transportation of the animals. In group 3, severe anaemia was present but also a leukopenia. For the different clinical chemical parameters measured, all three groups showed changes, explained mainly by the altered activity of enzymes induced by trace element deficiencies and imbalance. Impaired carbohydrate and lipid metabolism was seen in groups 1 and 3, with increased concentrations of glucose, lactate and triglycerides in serum. Increased concentrations of total bilirubin were measured in all three groups (bile stasis was also seen post mortem). A considerably increased concentration of serum urea was found in group 3, although there were no indications of renal insufficiency or dehydration. Regarding hormones, a substantial decrease was seen in thyroxine (T4) in group 3 as a result of the molybdenosis, but a minor decrease was also seen in group 1. Insulin on the other hand showed increased levels in group 3--and especially in group 2 due to the Cr deficiency but also affected by the molybdenosis. As could be expected, Cu deficiency (groups 1 and 3) caused low levels of caeruloplasmin, secondarily affecting the Fe metabolism in these animals. Protein abnormalities, detected as altered electrophoretic patterns of serum proteins, were also seen mainly in group 3. The findings were also confirmed by multivariate data analysis, where PCA revealed the overall impact of the deficiencies, and PLS regression coefficients indicated the influence on the various analytes. PMID- 10813455 TI - Environmental variables in a holistic evaluation of land contaminated by historic mine wastes: a study of multi-element mine wastes in West Devon, England using arsenic as an element of potential concern to human health. AB - Unusual and unexpected concentrations of a number of elements were identified in samples of house dust, that indicated the presence of mine wastes in an area where they were not expected. In pursuing this matter, several other very unusual observations and practices, involving highly contaminated mine wastes, were also identified. Here, using an available, but not a custom-made database, the matter is pursued. In England and Wales, the usual framework within which hazards are assessed, starts with an identification of those levels of exposure to elements which are considered to be safe and acceptable. At the other extreme, levels that are considered to be hazardous are identified together with procedures for dealing with them for different situations; for example, the manner in which contaminated land is to be used. The level at which an identification of sites and their use rests with the Local Authority, such as District Councils. Although new legislation is pending; at present for the non-occupationally exposed population there are no firm values to define which levels are acceptable and those that are not acceptable. One of the elements in the samples is arsenic and, because of its well-known toxicity, this element is selected to be of prime concern. However, simultaneous exposure to the general public is from a number of other elements, such as copper, lead, zinc, antimony, molybdenum, tin, selenium and mercury which are present in the mine wastes. The collective impairment to human health, if it should occur, is far too complex to evaluate, hence a need to focus attention upon arsenic which by any standard is present in some samples at very high concentrations, for example > 1000 mg/kg. Irrespective of any changes in permitted levels of exposure to the general public in the study area, together with those that are occupationally exposed, it is the past exposure that will give rise to the present patterns of morbidity and mortality. Because of a latent period between exposure and effects for the induction of cancers, of between 5 and 20 years, past exposures cannot be ignored. They need to be evaluated before changes are made in legislation. In England and Wales, concern to health is, in practice, invoked when there is clinical evidence of harm. With the exception of a few accidents in the study area of SW England, this is not identified, hence it is accepted, in practice, to conclude that no harm accrues following acute or chronic exposure to the mine wastes. There are reasons for questioning this, but if eventually supported, then the current very high costs for remediation of land may not be necessary and brown field sites can be used for a number of purposes. The primary concern is the lack of adequate scientific and clinical data, in relation to exposure to the wastes for the past 100 years or so. For many elements, compounds and substances, the general public is being made aware of potential risks to health but often the levels are extremely low. Using basic geochemical data for any area, it is possible to evaluate the expected background levels for elements. They should serve to identify levels that are acceptable, i.e. it is impractical or uneconomic to reduce them. Within the environment, simple tests are also available that can be used to rank risks to fauna and flora. There are also well-structured clinical evaluations of harm to humans available, which can also be ranked. All three need to be considered in the establishment of safe levels of exposure. It may not be practical to identify universal levels for exposure, rather each site, area or region needs to be considered separately in order to rank the potential risks. For the study area the exposures can be extremely high; compared with the general population, the number who are exposed is probably small. It is surprising that no effects upon human health have been noted. PMID- 10813456 TI - The Biological System of the Elements (BSE). Part II: a theoretical model for establishing the essentiality of chemical elements. The application of stoichiometric network analysis to the biological system of the elements AB - Stoichiometric Network Analysis (SNA), originally developed by the Canadian chemist Bruce L. Clarke during the 1970s, provides a most efficient means of reducing the background topology of complex interaction networks to some skeleton topology around which systems dynamics can be understood without jeopardising insight into complex dynamics by over- or miss-simplification. Since it focuses on the corresponding autocatalytic (AC) features of a feedback system as those which control overall behaviour to some extent, SNA deals with reaction kinetics in and beyond chemistry, e.g. with nuclear reactions. It is therefore quite straightforward to apply this manner of simplification, which in turn is supported by a number of mathematical theorems on systems behaviour and properties of AC cycles, to biological systems although their 'full' complexity may not even be assessed in the yet rare cases of complete genetic sequencing. Assuming there is a relationship between the kinds of metal or metalloid species and key biological/biochemical transformations to be promoted with their aid- this relationship being the subject of bio-inorganic chemistry--and that biochemistry is, in effect, about systems which can reproduce and thus behave autocatalytically, one can expect SNA to yield formally sound statements on basic features of biology and biochemistry too. If we sum up the facts and considerations concerning essentiality or possible essentiality in a biological system of elements (Markert, 1994), this means joining the triangular representation of BSE, including statements on (the degree of biological) evolution and aggregation levels, to SNA treatment of autocatalysis within hierarchical systems from metalloenzymes to entire biocoenoses. Arguments using preferred cluster sizes and aggregation tendencies from coordination chemistry are then employed to circumscribe possible functions within the BSE. They are also extended to metals hitherto not known to be essential, such as tellurium or scandium. PMID- 10813457 TI - Deposition and disease: a moss monitoring project as an approach to ascertaining potential connections. AB - In the years 1995 and 1996 the atmospheric deposition of elements in the EuroRegion Neisse (ERN) was determined in a biomonitoring project using mosses. The mosses Pleurozium schreberi and Polytrichum formosum were chosen as biomonitors because of their wide distribution in the area studied. The moss samples were analysed by ICP-MS and ICP-OES for their concentrations of 37 chemical elements. The results were shown in the form of maps. The data from the moss monitoring project served as a basis for determining those elements in the deposited material that promote the occurrence of disease. This was done by correlating the figures for the various diseases with the appropriate element concentrations in the mosses. Indications were found that a connection exists between the thallium content of mosses and the occurrence of cardiovascular disease and between Ce, Fe, Ga and Ge levels in the mosses and the incidence of diseases of the respiratory system. PMID- 10813458 TI - Critical examination of trace element enrichments and depletions in soils: As, Cr, Cu, Ni, Pb, and Zn in Swiss forest soils. AB - The aim of this study was to obtain an overview of trace element concentrations in Swiss forest soils and to critically assess the measured values with respect to anthropogenic input vs. lithogenic background. Twenty-three sites were selected which represent a broad range of natural forest sites, bedrock material and soil types of Switzerland. At each site, samples were collected from all genetic soil horizons down to a C or B/C horizon. Total concentrations of As, Cr, Cu, Ni, Pb, and Zn in all samples were determined by X-ray fluorescence spectrometry. There were distinct differences in the geological background values estimated from the concentrations measured in the samples from the lowest soil horizon. Background concentrations for Cr and Ni were lowest in granite and gneiss, whereas Pb and Zn were highest in limestone and marl. Enrichment or depletion of the trace elements was assessed using Zr as reference element. Within the same profile, the six trace elements showed completely different enrichment/depletion patterns with depth. The various natural processes and anthropogenic inputs that can lead to these patterns are critically discussed. Based on this critical assessment, pollution of the investigated forest soils was found to be most severe for Pb and Zn and to a somewhat lesser extent for As and Cu, whereas anthropogenic input of Cr and Ni seems to be less important. The data suggest that a critical evaluation of enrichment factors is a better tool to assess soil pollution with trace elements than the use of maximum allowable concentrations (MAC) for topsoil samples. The enrichment factors calculated as described here consider the effects of geological variation on metal abundances whereas the MAC does not. In order to obtain an estimate of soil solution concentrations, water extracts of the samples collected from a subset of 10 soil profiles were analyzed for the same trace elements. Solubility of all elements generally decreased with soil depth. An exception was Cr, Cu, and Ni solubility in the humus layer, which was lower than in the underlying mineral horizon. For all elements, solubility was higher for the collective of soil samples depleted in this element when compared to the samples, in which the element was enriched. PMID- 10813459 TI - A new approach for quantifying cumulative, anthropogenic, atmospheric lead deposition using peat cores from bogs: Pb in eight Swiss peat bog profiles. AB - Peat cores taken from eight Swiss peatlands were used to calculate inventories of anthropogenic Pb using either Sc or Zr to quantify Pb derived from rock weathering. The shapes of the Pb/Sc and Pb/Zr profiles suggest that Pb was supplied exclusively by atmospheric deposition at all sites. At one of the sites (Etang de la Gruere), anthropogenic Pb was calculated using both Sc and Zr as the conservative reference element. Lithogenic Pb determined using Sc was twice that obtained using Zr, possibly because Zr resides only in zircons which are dense compared to pyroxene and amphibole which are the main Sc-bearing phases in the earth's crust. However, the inventory of 'natural' Pb (supplied almost entirely by soil dust) is dwarfed by the anthropogenic inventory such that anthropogenic Pb calculated using Sc and Zr agree to within 5%. The total amount of anthropogenic Pb accumulated in the bogs was calculated by simply adding the mass of anthropogenic Pb for each peat slice over the length of each core. Cumulative, anthropogenic Pb calculated in this way ranged from 1.0 to 9.7 g/m2 and showed pronounced regional differences: the site south of the Alps (Gola di Lago in Canton Ticino) with direct exposure to the heavily industrialized region of northern Italy received nearly 10 times more anthropogenic Pb as the sites in more remote alpine regions (Schopfenwaldmoor in Canton Berne, and Mauntschas in Canton Grisons). The approach used here to calculate cumulative, anthropogenic, atmospheric Pb (CAAPb) is simple and robust, independent of the chronology of Pb deposition, and makes no assumptions about the immobility of Pb within the peat profile. Given the worldwide distribution of peat bogs, it should be possible to undertake continental and global inventories of atmospheric metal deposition, for both the natural and anthropogenic components of most trace metals of environmental interest. PMID- 10813460 TI - Arsenic in the geosphere--a review. AB - An attempt is made to quantify the global element cycle for arsenic, based on an extensive literature research with special emphasis on the most recent works. Reservoirs in and fluxes within and through lithosphere, atmosphere, pedosphere, hydrosphere, biosphere, and anthrosposphere are being presented. Crucial knowledge gaps are addressed and some simple model calculations partially question currently held ideas about sources, pathways, and the fate of arsenic in the environment. PMID- 10813461 TI - Environmental radioactivity: experience from the 20th century--trends and issues for the 21st century. AB - Environmental radioactivity has been an important area of research throughout the 20th century, with recent work having been stimulated mainly by: (i) the remarkable power of radionuclides as tracers of the rates and mechanisms of environmental processes and (ii) the potential health implications of contaminant radionuclides in the environment. A review is presented of salient aspects of environmental radioactivity, including consideration of tracer applications, sources and environmental impact of anthropogenic radionuclides, radioactive waste disposal and future exploitation of nuclear energy. PMID- 10813462 TI - Global outlook on nutrition and the environment: meeting the challenges of the next millennium. AB - As we enter the new millennium, nearly 800 million of the World's population will remain chronically malnourished. Nearly 200 million children are moderately to severely underweight, while 70 million are severely malnourished. And those who are yet to be born will be faced with the same set of circumstances that predispose them to malnutrition and its consequences. Eradication of nutritional deficiencies among women and children on a global scale are needed to ensure improved quality of life for the next generation of citizens. Primary deficiencies in vitamin A, iron, iodine, calcium, folic acid and trace elements such as zinc are compounded by pollutants caused by human activity. Environmental lead, arsenic, mercury, and other heavy metals that enter the food chain can seriously deplete body stores of iron, vitamin C and other essential nutrients leading to decreased immune defenses, intrauterine growth retardation, impaired psycho-social faculties and other disabilities associated with malnutrition. Increased susceptibilities to communicable diseases, and those provoked by water or insect borne vectors are additional risks encountered by malnourished individuals. Migration of populations from rural to urban centers and the expansion of major metropolitan areas have had a significant and adverse impact on the quality of life of these citizens. In the next 20 years most of the growth in urban populations will be in Asia and Latin America. Urbanization and the resultant burden on limited national resources is a major contributory factor to malnutrition. There are many other lifestyle-associated disabilities such as use of tobacco (cancer) and alcoholism that require active intervention. Within the family unit, socioeconomic factors and the status of women (literacy, economic independence) are major determinants of the quality of life. In the coming century, the World will have to meet these challenges by careful planning and international cooperation. PMID- 10813463 TI - Essentiality of selenium in the human body: relationship with different diseases. AB - In the present review different aspects related to the essential element selenium in the human organism are considered. A large number of human studies have been performed in order to improve knowledge on the influence of this element in the origin and development of several degenerative diseases. Selenium deficiencies among human beings as well as animals are being recognized worldwide to be related to a number of pathologies. This element has also the special characteristic that the range between its essential and toxic character is very close, and consequently daily dietary intake should be appropriately monitored in individuals. Nevertheless, nowadays there is still a lot of controversy about the optimum dietary level of this element in order to cure or to prevent the appearance of diseases such as cirrhosis, cancer, diabetes, or cardiovascular pathologies. Results obtained in several animal and epidemiological studies have indicated that Se could constitute a dietary factor with protective action against several degenerative diseases. PMID- 10813464 TI - Urban population exposure to lead and cadmium in east and south-east Asia. AB - Information is still scarce for many Asian countries on general population exposure to two potentially toxic heavy metals of lead (Pb) and cadmium (Cd). The present review on publications from this study group is intended to provide an overview of the non-occupational exposure to Pb and Cd among general populations in east and south-east Asia. During the period of 1991-1998, surveys were conducted in four cities in south-east Asia, five cities in mainland China, and two cities each in Japan and Korea. Peripheral blood, morning spot urine and 24-h food duplicate samples were collected from 20-50 non-smoking adult women in each survey site. The samples were digested by heating in the presence of mineral acids, and then analyzed by inductively-coupled plasma mass spectrometry for the metals in blood (Pb-B and Cd-B), in urine (Pb-U and Cd-U), and in food duplicate (Pb-F and Cd-F). Measures were subjected to statistical analysis with an assumption of log-normal distribution. Pb-B, Pb-U and Pb-F levels varied substantially among the 11 urban sites; GM distributed in ranges of 32-65 microg/l for Pb-B, 2.1-7.5 microg/g cr (creatinine) for Pb-U, and 7-32 microg/day for Pb-F. The same was also the case for Cd exposure parameters, the distribution ranges being 0.5-1.8 microg/l for Cd-B, 1.2-3.1 microg/g cr for Cd-U, and 5-32 microg/day for Cd-F. It appeared that the exposure to Pb and Cd was highest in Kuala Lumpur, and Tokyo + Kyoto, respectively, and lowest in Tokyo + Kyoto and in Manila. Additional surveys in Japan showed that Pb exposure was even lower in rural areas than in Tokyo + Kyoto. When compared with the values reported in the literature, Pb exposure levels among Asian populations appear to be similar to the levels in Europe and in the United States, whereas Cd exposure seems to be higher in Asia than in Europe. The contribution of the dietary route in Pb absorption was variable and was inversely related to the extent of air pollution, whereas Cd uptake was almost exclusively via the dietary route with little contribution of the respiratory route. PMID- 10813465 TI - Possible priorities for future research in the field of marine environmental pollution. AB - In this article future priorities and key areas for future research are identified. Comments and remarks upon strategies to establish priority topics are provided. They concern geographical area, environmental biomonitoring, ecoanalytics, relationships between environment and man, etc. All questions and suggestions presented in this article represent a personal viewpoint of the author, established on the basis of the cited references, personal experience and after discussion with several colleagues. PMID- 10813466 TI - Eating to your hearts delight. AB - Food and knowledge of food is an essential part of life. Knowledge about how to get enough and how to get it right is an essential part of all cultures. Since food is vital, nutrition policy is critical. Nutrition policy is about who should eat what, why, when and how in order to promote better health. This article deals with the politics of nutrition, which is about who eats what, why, when, how, and with what impact on their lives. For food, whether it is scarce or abundant, affects people unequally, hence food has always been a social concern. Regulating its supply has been a source of civilization. The state of nutrition is one of the most potent indicators of the state of society. Hence, food is also one of the oldest objects of politics--indeed, one cannot think of politics without it. This is the first issue I will address: by offering a capsule history of humanity from the perspective of the politics of nutrition. Next, I will address, in a similarly compressed history, the development of the science of nutrition, as nutrition policy has become based on research--particularly on medical science. Finally, I will address nutrition policy as a subpart of the politics of nutrition--how it has been defined and how it has developed, its changing agendas and current concerns. PMID- 10813467 TI - Dracunculiasis eradication: delayed, not denied. AB - By the end of 1998, Asia was free of dracunculiasis (Guinea worm disease), with Pakistan, India, and Yemen having interrupted transmission in 1993, 1996, and 1997, respectively. Transmission of the disease was also interrupted in Cameroon and Senegal during 1997. Chad reported only 3 cases during 1998. Dracunculiasis is now confined to only 13 countries in Africa. The overall number of cases has been reduced by more than 97% from the 3.2 million cases estimated to have occurred in 1986 to 78,557 cases reported in 1998. Because the civil war in Sudan remains the major impediment to eradication of dracunculiasis, the interim goal is to stop all transmission outside that country by the end of 2000. The most important operational need now is for national programs to improve the frequency and quality of supervision of village-based health workers in order to enhance the sensitivity of surveillance and effectiveness of case containment. PMID- 10813468 TI - Short report: study on the efficacy of a new long-acting formulation of N, N diethyl-m-toluamide (DEET) for the prevention of tick attachment. AB - The beneficial effects of N, N-diethyl-m-toluamide (DEET) against biting insects of human and animals appear to last <6 hr after a single application to the skin. To prolong the repellent effects of DEET, recently we developed a new long-acting formulation of DEET called LIPODEET. This preparation was retained in the skin for a longer duration of time with minimal systemic absorption. In this study, we have evaluated the protective effect of three compounds (DEET, LIPODEET, and Morpel 220) against attachment of two species of ticks (Amblyomma americanum and Dermacentor variabilis) to rabbit ears. Results show that LIPODEET and Morpel 220 were highly effective in preventing tick attachment to the skin for a longer duration of time (up to 72 hr) than DEET after a single application. Moreover, LIPODEET was found to be acaricidal to both the species of ticks. PMID- 10813469 TI - Economic impact of febrile morbidity and use of permethrin-impregnated bed-nets in a malarious area I. Study of demographics, morbidity, and household expenditures associated with febrile morbidity in the Republic of Benin. AB - In preparation for a study on the effect of bed net use on malaria, this article describes febrile morbidity and malaria expenditures in a sub-Saharan area (Benin) of hyperendemic malaria. The 325 randomly selected households were visited weekly between April 1994 and March 1995 to determine febrile morbidity and household expenditures for prevention and treatment. The results indicate that rural children had two febrile episodes annually compared with 0.3 episodes among children living in the city. There was no difference in mean annual febrile episodes between adults and children (adults = 1.5, children = 1.5; P = 0.48) and in the expenditures per febrile episode (adults = US$1.85, children = US$1.62; P = 0.45). Annual prevention expenditures were higher for adults than for children (US$1.73 and US$1.28, respectively; P < 0.001), although there was no significant difference in expenditures for annual treatment for adults and children (US$2.15 and US$2.34, respectively). These and other findings are analyzed further and discussed. PMID- 10813470 TI - Economic impact of febrile morbidity and use of permethrin-impregnated bed nets in a malarious area II. Determinants of febrile episodes and the cost of their treatment and malaria prevention. AB - The objective of this study is to determine the effect of permethrin insecticide treated bed net (PITN) use on the incidence of febrile episodes and on household malaria expenses in Benin. Over the course of one year, 208 randomly selected PITN user and non-user households were visited weekly to determine expenditures on febrile morbidity and its treatment, and to monitor spending on malaria prevention. Multivariate analyses were performed to distinguish the effects of PITN use from other important determinants of morbidity, such as malaria-related beliefs and practices, income, and other socio-economic variables. Results from the logistic regression analysis show that the use of PITNs decreases the risk of febrile episodes by 34% in children living in the rural zone. Multiple regression analysis reveals that PITN use does not reduce prevention and treatment expenses. These expenses are significantly associated with women's income. This report also discusses other factors associated with febrile morbidity and malaria-related expenditures. PMID- 10813471 TI - Short report: phylogenetically distinct hepatitis E viruses in Pakistan. AB - Hepatitis E, which is enterically transmitted, is the most common cause of acute hepatitis in much of Asia. Phylogenetic analysis of several isolates of hepatitis E virus (HEV) from Asia suggests that transmission of this virus is geographically restricted. In Sarghoda, Pakistan, HEV Sar-55 was isolated from a 1987 outbreak. It belongs to the Central-Asian cluster of the Asian sub-genotype. We now report the complete sequence of a second Pakistan HEV from a 1988 outbreak in Abbottabad. The Abbottabad nucleotide sequence was compared with 15 other complete HEV sequences using statistical methods of phylogenetic analysis. The analysis showed that Abbottabad HEV belongs to the South Asia cluster of the Asian sub-genotype. The sequence differences of the 2 Pakistan isolates recovered only one year apart suggest that HEV of 2 distinct origins circulate in Pakistan. PMID- 10813472 TI - Chloroquine-resistant isolates of Plasmodium falciparum with alternative CG2 omega repeat length polymorphisms. AB - A particular polymorphism in the cg2 gene has previously been linked to chloroquine resistance in reference isolates of Plasmodium falciparum. To assess the association of this polymorphism with chloroquine resistance in field specimens of P. falciparum, we analyzed the omega repeat region of the cg2 gene in 47 isolates of P. falciparum collected in the Ingwavuma District of northern KwaZulu-Natal, South Africa. Polymerase chain reaction (PCR) primers, which were designed to amplify the region of DNA surrounding the omega repeat, were used to obtain omega repeat PCR products from the field isolates. The PCR product for each isolate varied in length, depending on the number of cg2 omega repeats for that isolate. We found that several in vivo and in vitro chloroquine-resistant isolates of P. falciparum did not have the expected 16 omega repeats. These results suggest that the link between the cg2 polymorphism and chloroquine resistance identified previously may not apply in all malarious areas. PMID- 10813473 TI - Differentiation of Aedes triseriatus (Say) from Aedes hendersoni Cockerell (Diptera: Culicidae) by restriction fragment length polymorphisms of amplified ribosomal DNA. AB - Aedes triseriatus is the primary vector of LaCrosse (LAC) virus, which can cause encephalitis, especially in young children. Aedes hendersoni, a sibling species of Ae. triseriatus, has a salivary gland barrier to LAC virus and, therefore, is not considered a vector of this virus. Adults of Ae. triseriatus are morphologically indistinguishable from those of Ae. hendersoni, and the two species are sympatric in the eastern United States. A definitive method of identifying field specimens is an important part of any disease surveillance program, particularly in the case of LAC virus. This study identifies restriction enzymes that produce species-specific restriction fragment length polymorphisms (RFLPs) from amplified ribosomal (r) DNA. In addition, sequences of the internal transcribed spacers 1 and 2 and the 5.8S regions of the rDNA were used to confirm the RFLP patterns. This study is the first to compare nucleotide sequences from Ae. triseriatus and Ae. hendersoni. PMID- 10813474 TI - Genetic isolation by distance among Aedes aegypti populations along the northeastern coast of Mexico. AB - A population genetic analysis of gene flow was conducted among 10 Aedes aegypti collections from seven cities along the northeastern coast of Mexico. Four collections were made from Monterrey to examine local patterns of gene flow. Markers included 60 random amplified polymorphic DNA (RAPD) loci amplified by the polymerase chain reaction and single strand conformation polymorphism analysis of variation in a 387-basepair region of the NADH dehydrogenase subunit 4 from the mitochondrial DNA (mtDNA). Seven mitochondrial haplotypes were detected and phylogenetic analysis identified two well-supported clades. Regression analysis of geographic distances and pairwise FST estimated from RAPD markers indicated that populations are isolated by distance and that free gene flow occurs among collections within 90-250 km. Isolation by distance was not detected using mtDNA haplotypes. The Nuevo Laredo collection had unique RAPD and mtDNA haplotype frequencies and reduced heterozygosity suggesting that few mosquitoes established this population. PMID- 10813475 TI - Gametocytemia and infectivity to mosquitoes of patients with uncomplicated Plasmodium falciparum malaria attacks treated with chloroquine or sulfadoxine plus pyrimethamine. AB - Plasmodium falciparum gametocytemia and its related infectivity for mosquitoes was studied in 115 patients (median age = 18 years, range = 4-45) with simple malaria attacks who lived in the hypoendemic area of Dakar, Senegal. Patients were included in a 28-day in vivo sensitivity test after treatment with chloroquine (CQ, n = 82) or sulfadoxine plus pyrimethamine (SP, n = 33). The prevalence of resistant infections was 58.5% in those treated with CQ and 0% in those treated with SP. The gametocytemia peaked at day 7 after treatment. The maximal gametocyte prevalence was 38.2% in the CQ-sensitive infection group, 89.6% in the CQ-resistant group, and 97.0% in those treated with SP The maximal geometric mean gametocytemia was 2.19/microl in the CQ-sensitive infection group, 29.12/microl in the CQ-resistant group and 85.55/microl in those treated with SP. The period between appearance of the first clinical symptom and treatment was positively related to gametocyte prevalence at days 0 and 2. Experimental infection of wild Anopheles arabiensis using membrane feeders was performed at days 0 and 7, and mosquito infectivity was measured by oocyst detection on the midgut. At day 0, 14.1% of the patients had infected at least 1 mosquito, and at day 7, this value was 38.5%. The mean percentage of infected mosquitoes was 3.2% at day 0 and 12.6% at day 7. At day 7 after treatment with CQ, the relative risk for patients with resistant infections of infecting anophelines was 4.07 higher than in those with sensitive infections. No difference was observed in infectivity for mosquitoes between RI-type resistance and the RII + RIII-type resistance. A sporonticidal effect of SP was observed at day 7 after treatment. These data show that P. falciparum gametocytes and their infectivity for mosquitoes were differentiated according to the drug used, its efficacy, and the duration of symptoms before treatment; they were not dependent on the density of asexual stages. Prompt treatment of malaria cases performed at the beginning of symptoms could limit the spread of resistant parasites. PMID- 10813476 TI - Aedes aegypti in Tahiti and Moorea (French Polynesia): isoenzyme differentiation in the mosquito population according to human population density. AB - Genetic differences at five polymorphic isoenzyme loci were analyzed by starch gel electrophoresis for 28 Aedes aegypti samples. Considerable (i.e., high Fst values) and significant (i.e., P values >10(-4)) geographic differences were found. Differences in Ae. aegypti genetic structure were related to human population densities and to particularities in mosquito ecotopes in both Tahiti and Moorea islands. In highly urbanized areas (i.e., the Papeete agglomeration), mosquitoes were highly structured. Recurrent extinction events consecutive to insecticidal treatments during dengue outbreaks tend to differentiate mosquito populations. In less populated zones (i.e., the east coast of Moorea and Tahiti), differences in ecotope characteristics could explain the lack of differentiation among mosquitoes from rural environments such as the east coast of Tahiti where natural breeding sites predominate. When the lowest populated zones such as Tahiti Iti and the west coast of Moorea are compared, mosquito are less differentiated in Moorea. These results will be discussed in relation to the recent findings of variation in mosquito infection rates for dengue-2 virus. PMID- 10813477 TI - Random distribution of mixed species malaria infections in Papua New Guinea. AB - Plasmodium falciparum (Pf), P. vivax (Pv), P. malariae (Pm), and P. ovale (Po) infections are endemic in coastal areas of Papua New Guinea. Here 2,162 individuals living near Dreikikir, East Sepik Province, have been analyzed for complexity of malaria infection by blood smear and polymerase chain reaction (PCR) diagnoses. According to blood smear, the overall prevalence of Plasmodium infection was 0.320. Most individuals (0.283) were infected with a single species only. The prevalence of mixed species infections was low (0.037). Further analysis of a 173-sample subset by nested PCR of small subunit ribosomal DNA resulted in an overall 3.0-fold increase in prevalence of infection, with a 17.5 fold increase in the frequency of mixed species infections. Among mixed species infections detected by PCR, the frequency of double species was 0.364, and that of triple species was 0.237. Nine individuals (0.052) were infected with all 4 species. To determine if infection status (uninfected, single, and multiple infections) deviates from an independent random distribution (null hypothesis), observed versus expected frequencies of all combinations of Plasmodium species infections, or assemblages (Pf-, Pv-, Pm-, Po-, to Pf+, Pv+, Pm+, Po+), were compared using a multiple-kind lottery model. All 4 species were randomly distributed whether diagnosed by blood smear or PCR in the overall population and when divided into age group categories. These findings suggest that mixed species malaria infections are common, and that Plasmodium species appear to establish infection independent of one another. PMID- 10813478 TI - Pseudomonas (Burkholderia) pseudomallei in Thailand, 1964-1967: geographic distribution of the organism, attempts to identify cases of active infection, and presence of antibody in representative sera. AB - The purpose of this study, initiated in 1964 and concluded in 1967, was to define the distribution of Pseudomonas (now Burkholderia) pseudomallei in Thailand, to evaluate its importance as an etiologic agent, and to survey the presence of antibody in people that might indicate prior infection and/or contact with the microorganism. PMID- 10813479 TI - First field evidence for natural vertical transmission of West Nile virus in Culex univittatus complex mosquitoes from Rift Valley province, Kenya. AB - West Nile virus is a mosquito borne flavivirus endemic over a large geographic area including Africa, Asia, and the Middle East. Although the virus generally causes a mild, self-limiting febrile illness in humans, it has sporadically caused central nervous system infections during epidemics. An isolate of West Nile virus was obtained from a pool of four male Culex univittatus complex mosquitoes while we were conducting an investigation of Rift Valley fever along the Kenya-Uganda border in February-March 1998. This represents the first field isolation of West Nile virus from male mosquitoes and strongly suggests that vertical transmission of the virus occurs in the primary maintenance mosquito vector in Kenya. A phylogenetic analysis of the complete amino acid sequence of the viral envelope glycoprotein demonstrated a sister relationship with a Culex pipiens mosquito isolate from Romania made in 1996. This unexpected finding probably reflects the role of migratory birds in disseminating West Nile virus between Africa and Europe. PMID- 10813480 TI - The epidemiology of malaria in an epidemic area of the Peruvian Amazon. AB - A longitudinal study of malariometric indicators and their association with potential risk factors was conducted during August 1997-July 1998 at Padre Cocha, a village of 1,400 residents in the Peruvian Amazon. The incidence of Plasmodium falciparum infections during the study year was 166/1,000 persons; that of P. vivax was 826/1,000 persons. The mean duration of symptoms prior to diagnosis was 2 days; presenting geometric mean parasite densities were 3,976 parasites/microl for P. falciparum infections and 2,282 parasites/microl for P. vivax. There were no malaria-associated deaths. Consistent with the epidemic nature of malaria in the area, the incidence of both parasite species increased with age and there were no age-specific differences in mean parasite densities. No specific occupational risks for malaria were identified. Activities significantly associated with malaria risk reflected local vector behavior and included strolling outdoors after 6:00 PM and arising before 6:00 AM for adults, and attending evening church services for children. PMID- 10813481 TI - Hepatitis C virus genotypes in a northeastern area of Brazil. AB - We used a reverse transcription-polymerase chain reaction (RT-PCR) to obtain the genotypes of circulating hepatitis C virus (HCV) in patients from a Gastro Hepatology Unit in the city of Salvador (Bahia State) in northeastern Brazil. Viral RNA was detected in 83 (65.4%) of 127 anti-HCV seropositive serum samples. Positivity was significantly associated with alterations in levels of aspartate aminotransferase and alanine aminotransferase (P < 0.05). Genotyping of HCV was performed by RT-PCR using genotype-specific primers from the core region: 24.1% were infected with subtype 1a, 38.6% with 1b, 3.6% with 2, 21.7% with 3a, and 12.0% with a mixed genotype. There was no difference in genotype distribution when compared with results from other Brazilian locations. Surprisingly, the high frequency of genotype 3 in Brazilian samples continues to be different from that reported around the world and warrants further investigation. PMID- 10813482 TI - Analysis of Plasmodium vivax merozoite surface protein-1 gene sequences from resurgent Korean isolates. AB - The merozoite surface protein-1 (MSP-1) of Plasmodium vivax exhibits great antigenic diversity among different isolates of this parasite. This antigen is a useful genetic marker for studying the polymorphism of natural P. vivax parasite populations. One or more of these populations has been responsible for resurgent malaria now occurring in Korea. This paper reports the analysis of a highly polymorphic region between interspecies conserved blocks 5 and 6 of the MSP-1 gene, using the polymerase chain reaction to amplify the DNA fragment encompassing these regions from 25 Korean isolates, followed by sequencing. Almost all amino acid sequences of Korean isolates were nearly identical to that of Thai isolates TD525A (96.6-99.7%) and TD424 (96.3-99.5%), and very similar to that of the France-Belem strain when compared with other isolates (Sal-1, Sri Lanka, and Colombia). Interallelic recombination was found in the poly-Q repeat and a Sal-1 type amino acid structure was observed in all isolates. This study shows that the MSP gene nucleotide sequence of resurgent P. vivax in Korea is most similar to that of Thai isolates; however, the Korean strains are phylogenetically unique. PMID- 10813483 TI - Host factors affecting the delay of reappearance of Plasmodium falciparum after radical treatment among a semi-immune population exposed to intense perennial transmission. AB - To investigate host factors affecting the delay of reappearance of malaria parasites after radical treatment, a study was undertaken in Dielmo, Senegal, an area of intense perennial malaria transmission. A 7-day course of quinine was administered to 173 asymptomatic persons from 1 to 85 years of age and reappearance of malaria parasites in the peripheral blood was monitored weekly for 14 weeks. Additional thick blood films were made in case of fever as part of a daily clinical surveillance. The median times before reappearance of Plasmodium falciparum were 22, 39, and 53 days among persons 1-6, 7-14, and > or = 15 years of age, respectively (P < 0.0001). Multivariate analysis indicated that the daily rate of reappearance of P. falciparum was 2.2 (95% confidence interval [CI] = 1.2 4.5) times lower in sickle cell trait carriers than in AA individuals, and 1.5 (95% CI = 1.1-2.1) times lower in bed nets users than in non-users. The risk ratio for the daily risk of reappearance was significantly related to the level of parasitemia before treatment. No influence of glucose-6-phosphate dehydrogenase deficiency, HLA-B53, and DR13 were observed. Findings show that monitoring during a few weeks the reappearance of malaria parasites after treatment among a small cohort of individuals naturally exposed to malaria is relevant for investigating host resistance factors. This suggest that small, low cost, field trials may be very informative on the potential of new malaria vaccine candidates. PMID- 10813484 TI - Molecular epidemiology of malaria in Yaounde, Cameroon. VI. Sequence variations in the Plasmodium falciparum dihydrofolate reductase-thymidylate synthase gene and in vitro resistance to pyrimethamine and cycloguanil. AB - Pyrimethamine and cycloguanil, the major human metabolite of proguanil, are inhibitors of dihydrofolate reductase that play a key role in the treatment and prevention of chloroquine-resistant Plasmodium falciparum infections in sub Saharan Africa. Resistance to these antifolate drugs has emerged in some areas of Africa. Earlier molecular studies have demonstrated that point mutations at key positions of the dihydrofolate reductase-thymidylate synthase gene are strongly associated with antifolate resistance. However, whether the same or distinct mutations are involved in the development of resistance to both pyrimethamine and cycloguanil has not been well established in naturally occurring P. falciparum isolates. In this study, the in vitro responses to both antifolate drugs were measured in 42 Cameroonian isolates and compared with the complete sequence of the dihydrofolate reductase domain of the gene (from 34 of 42 isolates) to analyze the genotype that may distinguish between pyrimethamine and cycloguanil resistance. The wild-type profile (n = 11 isolates) was associated with low 50% inhibitory concentrations (IC50s) ranging from 0.32 to 21.4 nanamole for pyrimethamine and 0.60-6.40 nM for cycloguanil. Mutant isolates had at least one amino acid substitution, Asn-108. Only three mutant codons were observed among the antifolate-resistant isolates: Ile-51, Arg-59, and Asn-108. The increasing number of point mutations was associated with an increasing level of pyrimethamine IC50 and, to a much lesser extent, cycloguanil IC50. These results support a partial cross-resistance between pyrimethamine and cycloguanil that is based on similar amino acid substitutions in dihydrofolate reductase and suggest that two or three mutations, including at least Asn-108, may be necessary for cycloguanil resistance, whereas a single Asn-108 mutation is sufficient for pyrimethamine resistance. PMID- 10813485 TI - Scorpion envenomation and serotherapy in Morocco. AB - A clinical and biologic study was conducted in Morocco to assess the efficiency of antivenom therapy for treating victims of scorpion stings. Epidemiologic and clinical data were collected from 275 patients envenomed by Androctonus mauretanicus mauretanicus and Buthus occitanus scorpions. Patients received antivenom or other drugs. Blood samples were collected at the time of hospital admission and 1 hr and 3 hr after treatment. Serum venom levels were quantified by using an ELISA. An association was found between clinical signs of envenoming and the level of venom in serum. Patients classified as grade II (moderate envenoming) had higher serum levels of venom level than patients classified as grade I (mild envenoming). At admission to the hospital, the mean venom concentration was not significantly different between the group not treated with antivenom, the group who received 2-5 ml of antivenom, and the group who received 10 ml of antivenom. A significant decrease in serum venom levels and an improvement in the clinical conditions were observed in patients administered 10 ml of antivenom. The lower decrease in serum venom levels in patients who received 2-5 ml of antivenom was due to lower doses of antivenom. No difference in the venom concentration was observed in patients who were not treated with antivenom. The absence of administration of antivenom increased the risk of developing clinical signs at the end of the hospitalization period. However, this risk was much higher when more than 1 hr elapsed between the time of the scorpion sting and the time of hospital admission. The results demonstrate that antivenom is effective in decreasing circulating venom and morbidity. Serotherapy is more efficient when given as soon as possible after envenomation and with adequate quantities of antivenom. PMID- 10813486 TI - Cerebrospinal fluid studies in children with cerebral malaria: an excitotoxic mechanism? AB - The pathogenesis of cerebral malaria is poorly understood. One hypothesis is that activation of microglia and astrocytes in the brain might cause the cerebral symptoms by excitotoxic mechanisms. Cerebrospinal fluid was sampled in 97 Kenyan children with cerebral malaria, 85% within 48 hr of admission. When compared with an age-matched reference range, there were large increases in concentrations of the excitotoxin quinolinic acid (geometric mean ratio cerebral malaria/reference population [95% confidence limits] = 14.1 [9.8-20.4], P < 0.001) and total neopterin (10.9 [9.1-13.0], P < 0.001) and lesser increases in tetra hydrobiopterin, di-hydrobiopterin, and 5-hydroxyindoleacetic acid. There was no change in tryptophan concentration. In contrast, nitrate plus nitrite concentrations were decreased (geometric mean ratio = 0.45 [0.35-0.59], P < 0.001). There was a graded increment in quinolinic acid concentration across outcome groups of increasing severity. The increased concentration of quinolinic acid suggests that excitotoxic mechanisms may contribute to the pathogenesis of cerebral malaria. PMID- 10813487 TI - Detection of DNA of nocturnally periodic Brugia malayi in night and day blood samples by a polymerase chain reaction-ELISA-based method using an internal control DNA. AB - An internal control was used in a polymerase chain reaction (PCR)-ELISA-based technique to detect the Hha I repeat of the filarial parasite Brugia malayi. A single microfilaria added to 200 microl of blood was reliably detected. The assay was evaluated on field samples from persons living in an area endemic for Anopheles-transmitted, nocturnally periodic B. malayi in central Sulawesi, Indonesia. Examination of night blood of 138 individuals for the presence of microfilariae by filtration revealed 44 microfilaria carriers. All microfilaria carriers were also positive in the PCR-ELISA and, in addition, 14 more samples were proven to contain parasite DNA. The sensitivity of both methods was compared on night and on day blood samples collected from 113 persons. Whereas 37 microfilaria carriers were identified by filtration of night blood, no microfilariae were observed in the corresponding day blood samples. The PCR-ELISA result was positive in all 37 night blood samples of microfilaria carriers and in an additional 13 night blood samples without microfilariae. Parasite DNA was detected in 31 day blood samples of microfilaria carriers and in 3 day blood samples of amicrofilaremic persons. Assuming a sensitivity of the PCR-ELISA on night blood of 100%, the sensitivity of night blood filtration is 74% and that of the PCR-ELISA on day blood is 68%. These data suggest that the described PCR ELISA method is capable of detecting infections with nocturnally periodic B. malayi in day blood samples. Therefore, this method may facilitate both the identification of endemic areas and the monitoring of control programs. PMID- 10813488 TI - Diagnostic and prognostic value of an immunofluorescent assay for melioidosis. AB - Melioidosis caused by Burkholderia pseudomallei is endemic in southeast Asia. The clinical manifestations range from wound infections to acute septicemia. In some cases, recurrence can also occur following complete recovery. Case fatality rates are high and a major factor is the delay in the culture and identification of the bacterium. An immunofluorescent assay (IFAT) using whole-cell antigen for the detection of total antibodies to B. pseudomallei was tested with 650 sera. Using a cut-off value of 1:80, 66 sera from culture-confirmed cases were positive with titers > or = 320. In another 523 sera from patients in which no other etiology could be found, 149 (23.4%) were positive. To monitor disease activity, persistence of antibody levels was investigated on 61 serial sera samples collected from 14 other confirmed cases on follow-up visits while on oral maintenance therapy. The IFAT demonstrated a reduction in titers in cases of localized infections, suggesting that either the infection was being resolved or arrested while septicemic patients maintained high IFAT titers on follow-up, suggesting the possibility of continuous sequestration of antigen from an intracellular source. PMID- 10813489 TI - Evaluation of rapid diagnostic tests for the detection of human immunodeficiency virus types 1 and 2, hepatitis B surface antigen, and syphilis in Ho Chi Minh City, Vietnam. AB - An evaluation of three new rapid diagnostic test kits for human immunodeficiency virus types 1 and 2 (HIV-1/2), hepatitis B surface antigen (HBsAg), and syphilis involved a two-phase comparison of rapid diagnostic assays using prospectively collected from hospitals and clinics in Ho Chi Minh City, Vietnam. After specificity and sensitivity testing, three new rapid diagnostic test kits were tested in parallel with six commonly used diagnostic test kits. The Determine HIV 1/2 test had fewer indeterminate or equivocal results than the Capillus HIV-1/HIV 2 or HIV Blot 2.2 tests. However, the Determine HIV-1/2 test yielded one false positive result when compared with the Serodia HIV, HIV Blot 2.2, and microparticle enzyme immunoassay (IMx) HIV tests. The Serodia HBsAg test yielded more false-negative results when compared with the Determine HBsAg diagnostic test kit. The results of the syphilis diagnostic tests evaluated in this clinical trial consistently agreed with those of the rapid plasma reagin test for syphilis. The Determine Syphilis Treponema pallidum (TP) test had three false positive results compared with the Serodia TP and the Serodia TP x particle agglutination (PA) tests, which had two false-positive results that were confirmed as negative by an ELISA. Application of these serologic tests within this comparative evaluation framework, using the World Health Organization alternative testing strategies, proved to be an effective way to determine serostatus related to HIV, hepatitis B, and syphilis. PMID- 10813490 TI - Detection of Francisella tularensis in infected mammals and vectors using a probe based polymerase chain reaction. AB - We investigated the use of a TaqMan 5' nuclease assay (5NA) directed against the Francisella tularensis outer membrane protein (Fop) gene and a polymerase chain reaction-enzyme immunoassay (PCR-EIA) directed against the tul 4 gene for detection of this organism in experimentally infected mice and in field-collected tick vectors. We also evaluated the use of specially formulated filter paper (FTA) for rapid sample preparation. The 5NA had a detection limit of 1 pg of genomic DNA (<100 colony-forming units) and could be completed within several hours. The PCR-EIA could detect 1 pg of genomic DNA and 10 attograms (ag) (22 copies) of cloned insert, but takes longer to perform. Both assays were genus specific, and successfully detected F. tularensis in mouse tissues (5NA) and in tick extracts (PCR-EIA). The FTA paper provided inexpensive, rapid, template preparation for the tick extracts, mouse tissues, and DNA obtained from clinical specimens. These probe-based assays have the potential to provide rapid, real time/high-throughput molecular diagnostics in field situations. PMID- 10813491 TI - Immunohistochemical characterization of the inflammatory infiltrate in placental Chagas' disease: a qualitative and quantitative analysis. AB - Chagas' disease, a systemic illness endemic to some regions of South America, is caused by the protozoan Trypanosoma cruzi. Transplacental infection may occur during any phase and cause fetal death. This study is the first to characterize the inflammatory cells in chagasic villitis by immunohistochemistry. Paraffin sections of 8 placentas with villitis by T. cruzi (4 live births and 4 stillbirths), as well as 8 control placentas without inflammation, were stained with hematoxylin and eosin, monoclonal antibodies for CD45RO, CD20, CD45RO/OPD4, CD8, HNKI, CD15, MAC387, and CD68 proteins, and a polyclonal antibody for S-100 protein. Quantification of positive cells was performed in 3 different high-power fields. In all cases of chagasic villitis, the inflammatory infiltrate was composed mainly of CD68+ macrophages, T lymphocytes, and a few natural killer cells. Among T cells, CD8+ cells outnumbered CD4+ cells in all placentas (CD4+:CD8+ ratios ranged from 0.04 to 0.38). B cells were absent or rare. In stillbirths, villitis was diffuse and severe with numerous T. cruzi, while in live births it was focal with few parasites. Other features that characterized villitis in stillbirths were 1) frequent trophoblastic necrosis, 2) presence of MAC387+ macrophages and CD15+ granulocytes attached to the sites of trophoblastic necrosis, 3) low CD4+: CD8+ ratios in most cases, 4) increased numbers of S-100 positive cells in the villous stroma. In conclusion, CD68+ macrophages and CD8+ T lymphocytes were the major cell population in villitis caused by T. cruzi. However, the pattern of inflammatory reaction differed between stillbirths and live births and was probably related to the number of parasites in the placental villi. PMID- 10813492 TI - Epidemiology 1, 2, 3: origins, objectives, organization, and implementation. AB - This supplement is a report on the Epidemiology 1, 2, 3 (EPI 1, 2, 3) investigation, its origins, evolution, and findings that were carried out over a period beginning in 1990 and ending in 1994 in Egypt. The large scope and size of the study, the largest to date on schistosomiasis in Egypt, was a rationale for publishing a supplement to document EPI 1, 2, 3 methods and results collectively in sufficient detail to serve as a reference for planning, designing, and analyzing future epidemiologic studies and evaluation of schistosomiasis control in Egypt. The 3 objectives of EPI 1, 2, 3 were to 1) determine the changing patterns of Schistosoma haematobium and S. mansoni, 2) investigate factors contributing to differences between villages in the Nile Delta, Middle Egypt, and Upper Egypt, and 3) investigate risk factors for morbidity. The objectives were addressed using standardized techniques, stool and urine examinations, clinical examinations (including abdominal ultrasound), and questionnaires on a selected sample of the populations of selected villages in 9 governorates in Egypt. PMID- 10813493 TI - Epidemiology 1, 2, 3: study and sample design. AB - Bad sample designs and selection bias have plagued studies on schistosomiasis, and as a result some believe that schistosomiasis is too focal, making it difficult to draw reliable samples. The Epidemiology 1, 2, 3 (EPI 1, 2, 3) sample design, although complex, demonstrates that sampling theory is readily applicable to epidemiologic studies of schistosomiasis. The EPI 1, 2, 3 sampling scheme was designed to achieve the smallest feasible standard errors given EPI 1, 2, 3 objectives and certain logistical constraints. The sample design is a multi-stage selection of villages (ezbas, which were stratified by size) and households within each of 9 purposely selected Egyptian governorates. Villages and households were systemically selected from census frames. The sampling of ezbas was especially difficult because of the lack of complete sampling frames and their wide variation in population size. Ultimately, ezbas were stratified by size and then randomly selected from each stratum. Sample sizes for villages and ezbas and individuals within ezbas were calculated based on EPI 1 and 2 objectives, respectively. No re-selection was made for non-respondents. A 20% subsample of the full sample was drawn for clinical and ultrasonographic examinations. The sample selected from individual governorates closely parallel the age structure of the 1986 census of the respective rural populations. Details of the study design and related methods are given below. PMID- 10813494 TI - Quality control for parasitologic data. AB - Accuracy of data is of paramount concern for all research. The task of providing objective assurances of accuracy of parasitologic data for a large, multi-center epidemiologic research project in Egypt (Epidemiology 1, 2, 3 [EPI 1, 2, 3]) presented a unique set of challenges undertaken jointly by the Ministry of Health's Qalyub Center for Field and Applied Research with technical assistance from Tulane University (New Orleans, LA). The EPI 1, 2, 3 project was part of large bilateral research program, the Schistosomiasis Research Project, undertaken jointly by the governments of Egypt and the United States. This paper describes the nature of the quality control system developed to accomplish this task, presents results and discusses the findings. PMID- 10813495 TI - The epidemiology of schistosomiasis in Egypt: methods, training, and quality control of clinical and ultrasound examinations. AB - Morbidity due to schistosomiasis was evaluated in a subpopulation of 14,000 of the randomized sample in the Epidemiology 1, 2, 3 Project. It was measured by using a standardized questionnaire for obtaining medical history and symptoms and by performing standardized physical and ultrasound examinations. Reported herein are descriptions of the methods and training and quality control efforts made to insure that the morbidity data was reliable and consistent when collected by 7 different teams in 9 different governorates. PMID- 10813496 TI - The epidemiology of schistosomiasis in Egypt: patterns of Schistosoma mansoni infection and morbidity in Kafer El-Sheikh. AB - This is a descriptive report of the Epidemiology 1, 2, 3 project in Egypt that made use of large probability sampling methods. These results focus on Schistosoma mansoni infection in the northern Nile Delta governorate of Kafr El Sheikh. A probability sample of 18,777 persons, representing the rural population of the entire governorate, was drawn. The sample was designed not to exclude villages based on location or presence of health care facilities and to include representation of the smaller ezbas or hamlets. The objective was to obtain detailed estimates on age- and sex-specific patterns of S. mansoni infection, and to provide a baseline for prospective studies. Stool specimens were examined by the Kato method. The estimated mean +/- SE prevalence of S. mansoni infection in the rural population was 39.3 +/- 3.3% in 44 villages and ezbas after weighing for the effects of the sample design. The estimated mean +/- SE geometric mean egg count per gram of stool (GMEC) was 72.9 +/- 7.3. Prevalence and GMEC varied considerably by village and ezba, with ezbas having a significantly higher prevalence. Villages and ezba-specific prevalence was strongly associated with GMEC (r2 = 0.61, P < 0.001). The prevalence of S. mansoni infection increased by age to 55.4 +/- 3.2% at age 16 without a significant change in the adult ages. There were no gender differences until age 6, after which males were consistently higher until middle age, when the differences converged. The age- and sex specific pattern of GMEC varied widely; however, when the GMEC data were collapsed into 5-year age groups, the GMEC peaked at 81.5 +/- 12.1 eggs/g in the 10-14-year-old age group. These estimates provide the basis for evaluating control measures for reducing prevalence, intensity of infection, and transmission. PMID- 10813497 TI - The epidemiology of schistosomiasis in Egypt: Menofia Governorate. AB - Health questionnaires and parasitologic examinations of urine and stool were performed upon a stratified random sample of 10,899 individuals from 1,537 households in 27 rural communities in Menofia Governorate in Egypt in 1992 to investigate the prevalence of, risk factors for, and changing pattern of infection with Schistosoma sp. in the governorate. A subset, every fifth household, or 1,480 subjects, had physical and ultrasound examinations to investigate prevalence of and risk factors for morbidity. The prevalence of S. mansoni ranged from 0.3% to 72.9% and averaged 28.5%. The geometric mean egg count was 81.3 eggs/gram of stool. Age-stratified prevalence and intensity of infection was 30-40% and 60-80 eggs/gram of stool from the age of 10 onward; males had higher infection rates and ova counts than females in all age groups > 10 years old. Schistosoma haematobium was rare, being consequential in only 1 community. Risk factors for S. mansoni infection were male gender; age > 10 years; living in smaller communities; exposures to canal water; history of or treatment for schistosomiasis or blood in the stool; detection of splenomegaly by either physical or ultrasound; and ultrasound-detected periportal fibrosis (PPF). The more severe grades of PPF were rarely (21 of 1,450 examinations) detected. Risk factors for morbidity, i.e., ultrasound-detected PPF, were similar to those for infection. Schistosoma mansoni has almost totally replaced S. haematobium in Menofia. The prevalence of S. mansoni in rural communities remains high and average intensities of infection are moderate. The association of morbidity with schistosomal infection was variable and is obviously markedly influenced by both the frequent use of antischistosomal chemotherapy in communities in Menofia and by the prevalence of complications from chronic viral hepatitis in the population: hepatomegaly did not correlate with infection; PPF and splenomegaly, however, were related to S. mansoni infection in both individuals and communities. PMID- 10813498 TI - The epidemiology of schistosomiasis in Egypt: Ismailia governorate. AB - A multi-stage stratified sample of 12,515 individuals from 1,941 households in 42 villages in the Ismailia governorate of Egypt were surveyed for schistosomal infection. A subset of 2,390 subjects were surveyed for morbidity by physical and ultrasonographic examination. The prevalence of Schistosoma mansoni infection in rural Ismailia was 42.9% and the geometric mean egg count (GMEC) was 93.3 eggs/gram of stool, with considerable variability between communities. Prevalence and intensity peaked in the 20-30-year-old age group and was higher in males than in females. Prevalence and intensity of S. haematobium was very low: 1.8% and 3.5 ova/10 ml of urine GMEC, respectively. Canal water exposure risk factors for S. mansoni infection were males bathing (odds ratio [OR] = 2.2), females washing clothes (OR = 1.9), and children playing or swimming (OR = 2.3). Presence of in house piped water supply and latrine lowered infection rates (P < 0.001 and P = 0.002, respectively). Histories of S. mansoni infection (OR = 1.6) or treatment (OR = 1.5) and blood in feces (OR = 3.5) were associated with infection. Hepatomegaly (16.0%) was more frequently detected than splenomegaly (3.6%) by physical examination, with both being more frequent in older age groups. Splenomegaly, but not hepatomegaly, was associated with presence of S. mansoni ova in stools (OR = 1.4) and the community burden of infection (P = 0.02). Ultrasonographically detected hepatomegaly, splenomegaly, and periportal fibrosis (PPF) were detected in 43.0%, 17.4%, and 39.7% of the subjects, respectively. The higher grades of PPF were rare. Ultrasonographically detected splenomegaly, not hepatomegaly, was associated with S. mansoni infection, community burden of infection, and PPF. Risk factors for PPF were the same as for S. mansoni infection. There was a marginal association of PPF with infection and none (P = 0.33) with the intensity of infection in individuals or in the community. We conclude that in rural Ismailia, S. haematobium infection is rare but the prevalence and intensity of infection with S. mansoni is high. The risk of infection is associated with environmentally detected factors and behaviors. Hepatosplenic morbidity attributable to S. mansoni infection is low, presumably because of the favorable effect of wide application of praziquantel therapy. PMID- 10813500 TI - The epidemiology of schistosomiasis in Egypt: Qalyubia Governorate. AB - The primary objectives of this study, carried out in Qalyubia Governorate in Egypt (south-central Nile Delta), were to continue tracking historical trends of infection prevalence of Schistosoma mansoni and S. haematobium, determine whether satellites (ezbas) of mother villages differed significantly with respect to schistosomiasis transmission, and to asses schistosomiasis-induced morbidity on a population basis using ultrasonography. Our study revealed that S. haematobium has virtually disappeared from Qalyubia governorate, and that S. mansoni prevalence continues to decline slowly (17% in 1991 compared with 19% in 1990). The prevalence of intestinal schistosomiasis was actually higher in the mother villages than in the ezbas of the same villages, indicating that prevalence based on surveys of villages alone did not (at least for Qalyubia) cause underestimates of true prevalence. (A mother village is the large village in an area that includes hamlets or ezbas. In many areas, the infection rate in ezbas is significantly higher than in the larger central village.) Ultrasonographic studies revealed that less than 3% of the population had stage 2 or stage 3 periportal fibrosis, commonly associated with chronic schistosomiasis mansoni. This low level of morbidity was consistent with earlier data from Qalyubia, which also showed a low level of S. mansoni-induced morbidity in this governorate. PMID- 10813499 TI - The epidemiology of schistosomiasis in Egypt: Gharbia Governorate. AB - Health questionnaires and parasitologic examinations of urine and stool were performed upon a stratified random sample of 14,344 individuals from 1,952 households in 34 rural communities in Gharbia Governorate of Egypt to investigate the prevalence of, risk factors for, and changing pattern of infection with Schistosoma sp. A subset, every fifth household, of 1,973 subjects had physical and ultrasound examinations to investigate prevalence of and risk factors for morbidity. Community prevalence of Schistosoma mansoni ranged from 17.9% to 79.5% and averaged 37.7%. The geometric mean egg count (GMEC) was 78.9 eggs/gram of feces. The prevalence and intensity of infection was 40-50% and 70-100 eggs/gram of feces in those > or =10 years of age. Schistosoma haematobium was detected in 5 of the 34 communities. The maximum infection rate was 2.8% and mean GMEC in the five communities was 2.1/10 ml of urine. The overall prevalence of S. haematobium in the governorate was 0.3%. Risk factors for infection with S. mansoni were male gender, an age >10 years, living in smaller communities, exposures to canal water, prior therapy for schistosomiasis, or blood in the stool (in children only). Morbidity detected by physical examination or ultrasonography did not correlate with S. mansoni infection in individuals with the exception of periportal fibrosis (PPF, odds ratio [OR] = 1.25). Periportal fibrosis was detected in more than half of the subjects by ultrasonography; 5.3% had grade II lesions and 1.0% had the most severe grade III changes. Risk factors for morbidity as manifested by ultrasonographically detected PPF were similar to those for infection. Periportal fibrosis had a negative relationship with abdominal pain (OR = 0.45) and hepatomegaly detected by physical examination and ultrasonography (ORs = 0.72 and 0.68), but it was associated with splenomegaly (ORs = 4.14 and 3.55). The prevalence of PPF, hepatomegaly, and splenomegaly increased with age. There was no relationship between community burden of schistosomiasis mansoni and any measurements of morbidity with the exception of splenomegaly detected by physical examination (r = 0.40). Schistosoma mansoni has almost completely replaced S. haematobium in Gharbia, which has a high prevalence and moderate intensity of S. mansoni infection. Periportal fibrosis was detected by ultrasonography in more than half of the subjects, and 1 in 16 had grade II and III lesions. The only relationship between PPF and other morbidity findings was its positive relationship with splenomegaly and negative association with hepatomegaly. Hepatic morbidity is common in communities in Gharbia but the role of schistosomiasis mansoni in this is uncertain. PMID- 10813501 TI - The epidemiology of schistosomiasis in Egypt: Fayoum Governorate. AB - Health questionnaires and parasitologic examinations of urine and stool were performed upon a stratified random sample of 7,710 individuals from 1,109 households in 21 rural communities in Fayoum Governorate, Egypt in 1992 to investigate the prevalence of, risk factors for, and changing pattern of, infection with Schistosoma sp. in the governorate. A subset, every fifth household, or 1,038 subjects, had physical and ultrasound examinations to investigate prevalence of, and risk factors for, morbidity. The prevalence of S. haematobium ranged from 0% to 27.1% and averaged 13.7%. The geometric mean egg count (GMEC) was 10.0 eggs/10 ml of urine. Age-stratified prevalence and intensity of infection were 18-25% and 10-15 eggs/10 ml of urine in those 5-25 years of age. Schistosoma mansoni were detected in inhabitants of 13 communities, but 78.5% of the infections were focally present in a group of 4 satellite hamlets around a single village. The overall prevalence of S. mansoni in the governorate was 4.3% and the GMEC was 44.0 ova/g of stool. Risk factors for infection with either species were male gender, an age <20 years, living in smaller communities, and exposures to canal water by males. Histories of burning micturation, blood in the urine, or prior schistosomiasis and reagent strip detected hematuria and proteinuria were risks for S. haematobium, but not for S. mansoni. Both urinary tract and higher grades of hepatic morbidity were rare. Obstructive uropathy was present in 6.3% of the subjects and was more common in males and older people. Ultrasonography-detected bladder lesions were present in 5.2% and correlated with S. haematobium only in younger subjects and in those with hematuria and proteinuria. The prevalences of hepatomegaly, splenomegaly, and periportal fibrosis (PPF) were associated with each other and increased with age and in males. Ultrasonography-detected hepatomegaly and splenomegaly were weakly associated with S. mansoni infections only in children. The prevalence of PPF was greater in the 4 communities with >25% S. mansoni infection rates in comparison with the 17 other villages and ezbas. Transmission of S. mansoni is focally well established in Fayoum, which also has the highest prevalence of S. haematobium in the governorates surveyed by the Epidemiology 1, 2, 3 Project. However, both urinary tract and hepatic morbidity are relatively rare in the governorate. This probably results from the long-standing schistosomiasis control program in Fayoum, which suppressed intensity more than prevalence of infection, leading to less community morbidity. PMID- 10813502 TI - The epidemiology of schistosomiasis in Egypt: Minya Governorate. AB - Risk factors, prevalence, and intensity of infection with Schistosoma sp. and prevalence and magnitude of morbidity caused by schistosomiasis was assessed in a stratified random sample of 16,433 subjects from 2,409 households in 33 rural communities in Minya Governorate, Egypt. The prevalence of S. haematobium ranged from 1.9% to 32.7% among the communities and averaged 8.9%. The average intensity of infection was a geometric mean egg count (GMEC) of 8.5 per 10 ml of urine and ranged from 1.6 to 30.9. Prevalence was maximum (18-20%) in those 10-20 years of age and higher in males than in females. Intensity of infection followed the same pattern. Infection with S. mansoni was present almost exclusively in a single village, confirming spread of this species up the Nile River and its focality in Minya. Risk factors for S. haematobium infection were an age from 11 to 20; male gender; males bathing in, women washing clothing or utensils in, and children swimming or playing in canals; and a history of, or treatment for, schistosomiasis. Recent history of burning micturition was associated with infection in children but not in adults, while a history of blood in urine correlated with S. haematobium infection in both age groups. Reagent strip detected hematuria and proteinuria were highly associated, particularly in children, with S. haematobium infection. The presence of hepatomegaly or splenomegaly on physical examination was not associated with S. haematobium ova in the urine. Hepatomegaly, as measured by ultrasonography in the midclavicular line or the midsternal line, or ultrasonography-detected splenomegaly were not present more frequently in infected subjects than in uninfected subjects. Schistosoma ova were not detected more frequently in urine of subjects with ultrasonography-detected periportal fibrosis than in the urine from subjects without this finding. Ultrasonography-detected urinary bladder wall lesions were detected in only 6 (0.3%) subjects and obstructive uropathy was observed in 54 (2.7%) subjects. The absence of an association between prevalence of urinary tract morbidity and S. haematobium infections was surprising. Two possible explanations are 1) that repeated chemotherapy has reduced the prevalence of urinary tract morbidity and 2) that morbidity was not being detected by the ultrasonographic operators. PMID- 10813503 TI - The epidemiology of schistosomiasis in Egypt: Assiut governorate. AB - In the Assiut, Egypt Epidemiology 1, 2, 3 investigation, a sample of 14,204 persons in 10 villages, 31 ezbas (satellite communities), and 2,286 households was drawn from a rural population of 1,598,607. Parasitologic examination of urine and stool were made for Schistosoma haematobium and S. mansoni, and physical and ultrasound examinations were made on a 20% subsample. The overall estimated prevalence of S. haematobium was 5.2 +/- 0.5 (+/- SE). This varied considerably by village and ezba, ranging from 1.5% to 20.9%, with ezbas having a slightly higher overall prevalence than villages. The overall estimated geometric mean egg count was 6.6 +/- 0.5 eggs per 10 ml of urine and was consistently low throughout the communities. Infection with S. haematobium was associated with age (peak prevalence of 10.6 +/- 1.5% in 15-19-year-old age group) males, children playing in the canals, a history of blood in the urine, and reagent strip positivity for hematuria and proteinuria. The prevalence of either hepatomegaly or splenomegaly detected by physical examination was low (4.0% and 1.5%, respectively). The prevalence of hepatomegaly determined by ultrasonography was substantially higher, 24.1%. The prevalence of periportal fibrosis (PPF) was 12.0%, but grade II or III PPF was present in less than 1%. Ultrasonography determined hepatomegaly, in both the midclavicular line and the midsternal line, increased by age to more than 30%. Periportal fibrosis was more common in the age groups in which infection rates were the highest. At the village and ezba level of analysis, the prevalence of hepatomegaly, splenomegaly, and PPF tended to be higher in communities having the highest prevalence of infection with S. haematobium. PMID- 10813504 TI - The epidemiology of schistosomiasis in Egypt: Qena governorate. AB - Qena is the southernmost governorate of Egypt included in the Epidemiology 1, 2, 3 national study. A probability sample selected 17,822 individuals from 2,950 households in 34 ezbas and 10 villages from a total rural target population of 1,731,252 (based on the most recent 1986 census of the population by the Egyptian Central Agency for Public Mobilization And Statistics). Parasitologic examination of urine and stool were made for Schistosoma haematobium and S. mansoni, respectively, and physical and ultrasound examinations were made on a 20% subsample. The overall estimated prevalence of S. haematobium was 4.8 +/- 0.7% (+/-SE) and geometric mean egg count (GMEC) was 7.0 ova per 10 ml of urine. Considerable variation in prevalence was observed between the villages and ezbas, ranging from 0.0% to 20%, with the smaller ezbas having a slightly higher overall prevalence. The age- and sex-specific patterns of S. haematobium showed typical peak prevalence in early adolescence, with males having a higher prevalence than females. A history of hematuria was associated with current infection (odds ratio = 3.6, 95% confidence interval = 2.32-5.63). Hepatomegaly and splenomegaly determined by physical examination present in 7.9% and 3.0%, respectively. Ultrasonography-determined hepatomegaly of the left liver lobe was found in 10.1%. Ultrasonography-detected hepatomegaly in both the left and right lobes increased in prevalence from approximately 5% in children to 15-20% in adults. The prevalence of ultrasonography-detected splenomegaly increased slightly with age. Grade III periportal fibrosis was detected in only 2 individuals in the sample. Bladder wall lesions and obstructive uropathy were also very infrequent. Other associations with these measures are given. Most villages and ezbas had an S. mansoni prevalence of less than 1%. The exception was Nag'a El-Sheikh Hamad, where the prevalence was 10.3 +/- 0.5% (GMEC = 57.4 +/- 2.6). Two other communities also had a prevalence >1% (Ezbet Sarhan and Kom Heitin). PMID- 10813505 TI - The epidemiology of schistosomiasis in Egypt: summary findings in nine governorates. AB - Health questionnaires and parasitologic examinations of urine and stool were evaluated from a stratified random sample of 89,180 individuals from 17,172 households in 251 rural communities in 9 governorates of Egypt to investigate the prevalence of, risk factors for, and changing pattern of infection with Schistosoma sp. in Egypt. A subset, every fifth household, or 18,600 subjects, had physical and ultrasound examinations to investigate the prevalence of and risk factors for morbidity. Prevalence of S. haematobium in 4 governorates in Upper Egypt in which it is endemic ranged from 4.8% to 13.7% and averaged 7.8%. The geometric mean egg count (GMEC) ranged from 7.0 to 10.0 ova/10 ml of urine and averaged 8.1. Age stratified prevalence of infection peaked at 15.7% in the 10-14-year-old age group and decreased to 3.5-5.5% in all groups more than 25 years of age. Age-stratified intensity of infection peaked at approximately 10.0 ova/10 ml of urine in the 5-14-year-old age groups and was about half that in all groups more than 25 years of age. Males had higher infection rates and ova counts than females in all age groups. Schistosoma mansoni was rare in Upper Egypt, being consequential in only Fayoum, which had a prevalence of 4.3% and an average intensity of infection of 44.0 ova/g of stool. Risk factors for S. haematobium infection were male gender, an age <21 years old, living in smaller communities, exposures to canal water; a history of, or treatment for, schistosomiasis, a history of burning micturition or blood in the urine, and reagent strip-detected hematuria or proteinuria. The more severe grades (II and III) of ultrasonography detected periportal fibrosis (PPF) were rare (15 of 906) in these schistosomiasis haematobia-endemic governorates. Risk factors for morbidity (ultrasonography detected urinary bladder wall lesions and/or obstructive uropathy) were similar to those for infection, with the exception that risk progressively increased with age. Subjects with active S. haematobium infections were 3 times as likely as those without active S. haematobium infections to have urinary tract morbidity. The prevalence of S. mansoni in 5 governorates in Lower Egypt, where it is endemic, ranged from 17.5% to 42.9% and averaged 36.4%. The GMEC ranged from 62.6 to 93.3 eggs/g of stool and averaged 81.3. Age-stratified prevalence of infection peaked at 48.3% in the 15-19-year-old age group, but averaged 35-45% in all groups more than 10 years of age. The intensity of infection was highest in the 10-14-year-old age group, and showed a range of 70-85 eggs/g of stool in those > or =5 years of age. Males had higher infection rates and ova counts than females in all age groups. Schistosoma haematobium was rare in these governorates; Ismailia (1.8%) had the highest infection rate. Risk factors for S. mansoni were male gender, an age >10 years old, living in smaller communities, exposures to canal water, a history of, or treatment for, schistosomiasis or blood in the stool, detection of splenomegaly by either physical examination or ultrasonography, and ultrasonography-detected PPF. The more severe grades (II and III) accounted for 463 (13.3%) of the 3,494 having ultrasonography-detected PPF. Risk factors for morbidity (ultrasonography-detected PPF) were similar to those for infection except that inhabitants of smaller communities were not at increased risk. Active S. mansoni infection increased the odds ratio (OR) of having PPF by 1.37. In comparison with others with normal-size livers, subjects having hepatic enlargement in either the midclavicular line or the midsternal line detected by physical examination or ultrasonography had a reduced risk (ORs = 0.64-0.72) of PPF. The prevalences of lesions detected by ultrasonography were 23.7% for enlargement of right lobe of the liver, 11.3% for enlargement of left hepatic lobe, 20.6% for splenomegaly, and 50.3% for PPF. Schistosoma mansoni has almost totally replaced S. haematobium in Lower E PMID- 10813506 TI - Serum uric acid and risk of coronary heart disease: Atherosclerosis Risk in Communities (ARIC) Study. AB - PURPOSE: Approximately half of previous studies on serum uric acid have reported it to be an independent risk factor for coronary heart disease (CHD). We tested this hypothesis in the Atherosclerosis Risk in Communities (ARIC) Study. METHODS: A total of 13,504 healthy middle-aged men and women were followed prospectively for up to eight years. We identified 128 fatal and nonfatal CHD events in women and 264 in men. RESULTS: The age-, race-, and ARIC field center-adjusted relative risk of CHD for sex-specific quartiles of serum uric acid were 1.0, 1.39, 1.08, and 2.35 in women (p for trend = 0.009) and 1.0, 1.03, 0.89, and 1.21 in men (p for trend = 0.44), respectively. However, serum uric acid was correlated positively with many risk factors, and after multivariable adjustment, there was little evidence of an association of uric acid with CHD in either sex. CONCLUSIONS: Our results are not consistent with serum uric acid being an independent risk factor for CHD. PMID- 10813507 TI - A case control study of multiple myeloma at four nuclear facilities. AB - PURPOSE: Reported elevations of multiple myeloma among nuclear workers exposed to external penetrating ionizing radiation, based on small numbers of cases, prompted this multi-facility study of workers at US Department of Energy facilities. METHODS: Ninety-eight multiple myeloma deaths and 391 age-matched controls were selected from the combined roster of 115,143 workers hired before 1979 at Hanford, Los Alamos National Laboratory, Oak Ridge National Laboratory, and the Savannah River site. These workers were followed for vital status through 1990 (1986 for Hanford). Demographic, work history, and occupational exposure data were derived from personnel, occupational medicine, industrial hygiene, and health physics records. Exposure-disease associations were evaluated using conditional logistic regression. RESULTS: Cases were disproportionately African American, male, and hired prior to 1948. Lifetime cumulative whole body ionizing radiation dose was not associated with multiple myeloma, however, there was a significant effect of age at exposure, with positive associations between multiple myeloma and doses received at older ages. Dose response associations increased in magnitude with exposure age (from 40 to 50) and lag assumption (from 5 to 15 years), while a likelihood ratio goodness of fit test reached the highest value for cumulative doses received at ages above 45 with a 5-year lag (X2=5.43,1 df; relative risk = 6.9% per 10 mSv). Dose response associations persisted with adjustment for potential confounders. CONCLUSIONS: Multiple myeloma was associated with low level whole body penetrating ionizing radiation doses at older ages. The exposure age effect is at odds with interpretations of A-bomb survivor studies but in agreement with several studies of cancer among nuclear workers. PMID- 10813508 TI - The increasing marginal benefit of condom usage. AB - PURPOSE: Condom use is promoted as a primary strategy for preventing sexual transmission of human immunodeficiency virus (HIV). This paper analyzes how incremental changes in condom compliance rates can affect an individual's risk of acquiring HIV. METHODS: We developed a simple mathematical model of HIV transmission in which the cumulative probability of HIV infection depended in part upon the percentage of acts in which a condom was used. We applied basic methods of calculus to differentiate the mathematical model with respect to the probability of condom usage. We applied values from published studies to the model to illustrate how the marginal benefits of condom usage vary across different populations. RESULTS: In general, the marginal benefit of condom usage increases as condom compliance increases. CONCLUSIONS: The marginal benefits of increased condom usage vary across different risk groups and across different levels of condom compliance. These results offer insight into the motivation behind the decision of whether or not to use condoms, and indicate possible ways to optimize the use of resources devoted to increasing condom usage by at-risk populations. PMID- 10813509 TI - Immunization coverage among predominantly Hispanic children, aged 2-3 years, in central Los Angeles. AB - PURPOSE: To assess the immunization status of young children in a predominantly Hispanic region in and around downtown Los Angeles, and factors associated with complete immunization by age 24 months. METHODS: The information was gathered in a two-stage cluster survey with probability proportionate to estimated size (PPS) sampling of 30 clusters at the first stage, and simple random sampling of a constant number of children at the second stage. Vaccination coverage was determined by a review of the home immunization (HI) card, or of clinic records. RESULTS: Of the 270 sampled children, 91.5% were Hispanic and 6.7% were Black. Home telephone numbers were not available in 24.8% of the homes, and 34.1% reported having no health insurance. Vaccination coverage was over 90% for the first three doses of Diphtheria, tetanus toxoids and pertussis/ diphtheria, tetanus toxoids and acellular pertussis vaccine (DTP/DTaP)/Diphtheria and tetanus toxoids vaccine (DT), first two doses of poliovirus (Polio) vaccine, first dose of measles, mumps and rubella (MMR) vaccine, and first two doses of hepatitis B (Hep B) vaccine. Yet, by age 24 months, only 72.2% of the children had received the combined series of 4:3:1 (i.e., four DTP/DTaP/DT, three Polio, one MMR). This was further reduced to 64.4% for the combined series of 4:3:1:3:3 (i.e., four DTP/DTaP/ DT, three Polio, one MMR, three Haemophilus influenzae type b (Hib), three Hep B). Factors associated with completed on-time vaccination were having an HI card available during the interview and being enrolled in Supplemental Nutrition Program for Women, Infants and Children (WIC). CONCLUSIONS: While vaccination levels for individual antigens were found to be high, more emphasis needs to be placed on getting preschool children vaccinated on-time according to the Recommended Childhood Immunization Schedule. PMID- 10813510 TI - Development and validation of a calcium intake questionnaire for postmenopausal women in China. AB - PURPOSE: Develop and evaluate a food frequency questionnaire (FFQ) for assessing calcium intake in Chinese postmenopausal women. METHODS: An interviewer-conducted FFQ that asks respondents about their consumption of 110 calcium-containing foods over the previous week was developed and administered to 21 randomly selected postmenopausal women from Chengdu. This FFQ uses both the traditional weight estimation method of assessing portion size in China and a new volume-estimation method. Test-retest reliability was assessed by re-administering the questionnaire two weeks later and validity was assessed by comparing the computed daily calcium intake to that obtained by a researcher-conducted 4-day food record. RESULTS: Estimates of calcium intake derived from the volume-estimation method were significantly lower than estimates derived from the traditional weight-estimation method (median calcium intakes = 270 mg/day versus 570 mg/day), but were quite similar to those obtained from the 4-day food record (median calcium intake = 275 mg/day). The test-retest reliability and validity of the volume-estimation method (intraclass correlation coefficient (ICC) = 0.73 and 0.86, respectively) were better than those of the weight-estimation method (ICC = 0.58 and 0.39, respectively). CONCLUSIONS: The reliability and validity of the traditional weight-estimation FFQ method of assessing dietary intake of nutrients in China is poor; this method may significantly overestimate dietary intakes. The proposed volume-estimation FFQ method for assessing calcium intake is a simple, interviewer-conducted method that is both reliable and valid. The calcium intake of postmenopausal women in urban China is much lower than the recommended daily allowance of 800 mg. PMID- 10813511 TI - A study of smoking, p53 tumor suppressor gene alterations and non-small cell lung cancer. AB - PURPOSE: The purpose of this study was to investigate the relationship between smoking and p53 tumor suppressor gene alterations, and their association with clinicopathologic features and prognosis in non-small cell lung cancer (NSCLC). METHODS: For 111 of 119 stage I-III NSCLC patients that had been followed prospectively, tumor p53 protein accumulation was measured immunohistochemically (IHC). Staining was evaluated as a score (p53IHCS) combining intensity and percent distribution. RESULTS: Forty-eight of 111 (43%) tumors had p53IHCS > 1. p53 IHC was associated with increasing tumor size (T) (p = 0.035), nodal status (N) (p = 0.091), stage (p = 0.054), and histology: squamous cell carcinoma (70%) and adenocarcinoma (27%) (p = 0.0002). In logistic regression analysis, p53 IHC was associated with squamous cell histology versus other histotypes [adjusted odds ratio (OR)5.90, 95% confidence interval (CI) 2.34-14.90]. p53 IHC was not associated with smoking variables. In multivariate proportional hazards analysis, p53IHCS and pack-years smoked (PY), both as continuous variables, were negative prognostic factors. The adjusted hazard ratios (HR) for the survival outcome recurrence for p53IHCS and PY were 1.20 (95% CI 1.02-1.40) and 1.03 (95% CI 1.01 1.04), and for death due to recurrent disease (DRD) were 1.35 (95% CI 1.11-1.64) and 1.03 (95% CI 1.01-1.04), respectively. Comparing the 75th percentile to the baseline 0, the adjusted HR for p53IHCS (5 vs. 0) was 4.5 and for PY (55 vs. 0) was 5.1 for the outcome DRD. Both variables demonstrated a dose-response relationship with survival. CONCLUSIONS: PY and p53IHCS are significant, independent and important predictors of recurrence and DRD in stage I-III NSCLC. PMID- 10813512 TI - Preventable risk factors for nasal cancer. AB - PURPOSE: To determine preventable risk factors for cancers of the nasal cavity and paranasal sinuses in the United States, we analyzed data from the population based, case-control Selected Cancers Study. METHODS: Cases were men born between 1929 and 1953 who were diagnosed with primary nasal cancer between 1984 and 1988 and identified from population-based cancer registries; we narrowed the cohort to 70 subjects whose diagnosis of nasal cancer was confirmed by pathology review. All living controls interviewed for the Selected Cancers Study were included as the comparison group (n = 1910); they were recruited by random-digit dial telephone and were frequency-matched to the lymphoma cases of the Selected Cancers Study by geographic area and age. Both cases and controls were interviewed by telephone. RESULTS: Logistic regression analyses showed that cases were 2.5 times more likely than controls to have smoked cigarettes [95% confidence interval (CI) = 1.1-5.3], and 2.2 times more likely to have worked in selected occupations, including lawn care, forestry, and maintenance of highway right-of-way areas (CI = 1.2-3.7). These occupations may cause workers to be exposed to pesticides or herbicides. The population attributable risk (PAR) was 53% for having ever smoked cigarettes. CONCLUSIONS: The study results suggest that among U.S. men, some nasal cancer may be preventable by avoiding cigarette smoking. PMID- 10813513 TI - A comparison of melanoma mortality among WWII veterans of the Pacific and European theaters. AB - PURPOSE: To determine whether cutaneous melanoma is associated with past military service in tropical locations, as a test of the 'critical period' sunlight exposure hypothesis. METHODS: Mortality data from a longitudinal follow-up study of 5524 former prisoners of war (POW) and 3713 non-POW veteran controls, all white male veterans of World War II (WWII), were examined to determine whether death attributed to melanoma was associated with history of military service in Pacific or European theaters during WWII or with POW status. Deaths from colon cancer were used as a comparison outcome. RESULTS: During 50 years of follow-up, there were 18 deaths from melanoma and 83 deaths from colon cancer among the cohort. Melanoma mortality varied with theater of war and POW status, whereas colon cancer mortality was similar for all subgroups. Pacific War POWs were at the highest risk overall (odds ratio (OR), 3.35; 95% confidence interval (CI), 0.39-28.76), whereas veterans of the Pacific War had nearly the same risk of melanoma (OR, 1.04; 95% CI, 0.09-11.94) compared with veterans of the European War. European theater POWs had a higher risk than non-POW veterans (OR, 2.76; 95% CI, 0.31-24.81). None of these differences, however, were statistically significant. CONCLUSIONS: To the extent that POW status is associated with higher sun exposure, these data are consistent with the hypothesis that exposure to high levels of solar radiation in young adulthood is associated with a higher risk of melanoma mortality. PMID- 10813514 TI - Near-monodisperse sodium polymethacrylates: characterization by linear salt gradient anion-exchange chromatography AB - A family of six near-monodisperse homopolymers of sodium methacrylate (Mn = 1100, 3200, 5500, 7200, 14100, and 21000) is characterized by linear salt gradient anion-exchange chromatography. Although the retention times depend on the initial and final salt concentrations of the gradient, they are almost independent of the molecular weight of poly(sodium methacrylate). This suggests that anion-exchange chromatography is incapable of resolving mixtures of a given polyelectrolyte to their components of various molecular weights, and it is therefore impossible to identify the polydispersity of such a sample using this method. The independence of the retention times from molecular weight is also predicted by a theory based on stoichiometric mass-action ion-exchange. Using this theory and our experimental retention times, the equilibrium anion-exchange constant and the corresponding Gibbs free energy of anion-exchange of the monomer repeat unit are calculated to be around 2.1 and -1.8 kJ/mol, respectively. PMID- 10813515 TI - Taking into account both preparation and injection in high-performance liquid chromatography linearity studies AB - As mentioned in International Conference on Harmonisation publications, linearity is a principal parameter in method validation. The most popular statistical tool used is linear least-squares regression. Contrary to what is still very often practiced, the correlation coefficient can in no way be considered as an indicator of the fit quality. There is in fact a test called the "lack-of-fit test" that enables one to answer the question, "Is the linear model adapted to the calibration curve?". However, this test can give erroneous conclusions when, at each level, several sources of variation for the response are influent. It often occurs in high-performance liquid chromatography, as shown in a following example, where the calibration curve is obtained from repeated injections of repeated dilutions of a parent solution. The lack-of-fit test rejected linearity, although it was at least questionable. In fact, the reason for a discrepancy of this kind lies in the presence of a double source of variation: injection and dilution. It is possible to overcome the problem by mixing a nested ANOVA with the standard least-square linear regression. As shown in an example, implementing this methodology for data processing allows one not only to carry out an unbiased lack-of-fit test but also give estimates of the dispersion introduced respectively by the preparation and the injection. PMID- 10813516 TI - Prediction of isocratic nonaqueous reversed-phase high-performance liquid chromatography retention parameters and response factors of triacylglycerols detected by an ultraviolet-diode array-evaporative light-scattering on-line system. AB - Nonaqueous reversed-phase high-performance liquid chromatography of 10 homogeneous triacylglycerol molecular species (TAG), both saturated and unsaturated, is carried out. The eluate from the column is detected by an ultraviolet diode array detector (DAD) on-line with an evaporative light scattering detector (ELSD). The retention parameters (as selectivities, alpha) for 220 TAGs are determined, and the obtained values are related to the following structural parameters: total carbon number; mono-, di-, and triunsaturated fatty acid residues number/molecule; and monounsaturated fatty acid carbon number. Multiple regression analysis is carried out to obtain a relationship for the prediction of alpha values of any TAG when the same experimental conditions are used. In regard to the quantitative analysis of the separated TAG species, the dependence of response of the two on-line detectors on the aforementioned structural parameters is studied. Three different wavelengths (205, 210, and 215 nm) are considered for TAG detection by DAD; in each case, the obtained multiple regression model shows a good correlation between the dependent variable and predictive values of the TAG species (response factors and considered structural parameters, respectively). The ELSD gives responses exponentially related to injected amounts. Also, in this case, an attempt to relate the response factors of each considered detector to some structural parameters of TAG species is carried out. The results of this study are used to analyze the TAG fraction from an olive oil. PMID- 10813517 TI - Determination of fluoxetine, fluvoxamine, and clomipramine in pharmaceutical formulations by capillary gas chromatography. AB - A simple and fast capillary gas chromatographic method with flame ionization detection is proposed for the simultaneous determination of fluoxetine, fluvoxamine, and clomipramine without a prederivatization. The reported method is the first one that allows the determination of three selective serotonin reuptake inhibitors. Optimal conditions for the quantitative separation were investigated: column head pressure (80 kPa), injector and detector temperatures (260 and 250 degrees C), time and temperature for the splitless step (0.75 min and 60 degrees C), size of sample (2 microL), and oven temperature program, providing analysis times shorter than 10 min. Aspects such as the stability of the solutions, linearity, accuracy, and precision are examined in order to validate this method. Peak purity and detection and quantitation limits are also assessed using mass selective detection. The scope of the validated method is tested in the analysis of pharmaceutical preparations, with recoveries between 97.5 and 102.5% with regard to their nominal contents. PMID- 10813518 TI - Elimination of amino acid interferences in the chiral ligand-exchange chromatographic analysis of lactic acid enantiomers in wine. AB - Chiral ligand-exchange liquid chromatography is used to identify and quantitate lactic acid enantiomers in wines that have or have not undergone malolactic fermentation. The stationary phase is (R)-penicillamine, which is bound lipophilically to a C18 bonded silica matrix. The mobile phase is 1mM copper sulfate, and the detection mode is ultraviolet. Serious interference from (S) aspartic acid and other amino acids is eliminated by the use of propanesulfonic acid-type cation exchange solid-phase extraction cartridges prior to chromatographic analysis. Lactic acid enantiomers in wine are quantitated in the range of 10 to 500 mg/L. The detection limit is 3 mg/L. The method is also successful in the determination of lactic acid enantiomers in certain beers (e.g., lambic beers), kim-chi, sauerkraut, and various yogurts. PMID- 10813519 TI - Determination of selected herbicides and phenols in water and soils by solid phase extraction and high-performance liquid chromatography. AB - A high-performance liquid chromatography procedure or the determination of the herbicides simazine, propazine, bromacil, metoxuron, and hexazinone is elaborated. Stationary phases RP8 and RP18 and mixtures of methanol-water (2:1 and 1:1, v/v) as a mobile phase are applied for this purpose. The conditions for solid-phase extraction are established, allowing the separation of phenols and herbicides in their mixtures and the extraction of phenols (from river and coke plant water) and herbicides (from the soil samples). PMID- 10813521 TI - Gas chromatography problem solving and troubleshooting PMID- 10813520 TI - The paper of Vassilaros et al. published in the October 1999 issue PMID- 10813522 TI - Liquid chromatography problem solving and troubleshooting PMID- 10813523 TI - Freezing hematopoietic stem cells. PMID- 10813524 TI - Immunotherapy with dendritic cells: coming of age? PMID- 10813525 TI - Esmail D. Zanjani, Ph.D.. Interview by Vicki P. Glaser. PMID- 10813526 TI - A novel combination of paclitaxel, etoposide, and cyclophosphamide for stem cell mobilization and tumor cytoreduction in ovarian cancer. PMID- 10813527 TI - Kaposi's sarcoma-associated herpesvirus-encoded interleukin-6. AB - Since the discovery of the virus in 1994, the rapid pace with which Karposi's sarcoma-associated herpesvirus (KSHV) research has progressed has quickly led to a broad understanding of the structure of the virus and its biology and pathology in humans. Molecular piracy of potentially useful cellular genes has emerged as a characteristic feature of this virus. The viral homolog of human IL-6, vIL-6 is an example in kind. Studies in vitro and in vivo have shown that vIL-6 can stimulate the growth of KSHV-infected primary infusion lymphoma (PEL) cells, can promote hematopoiesis, and act as an angiogenic factor through the induction of vascular endothelial growth factor (VEGF). It is not difficult to envision how vIL-6, through these properties and perhaps others yet to be identified, can contribute to KSHV survival and spread in the human population. PMID- 10813528 TI - Immunologic methods of purging in autologous stem cell transplantation. AB - Malignant cells in the stem cell product have been shown to contribute to disease recurrence in patients who relapse after autologous transplantation. Immunologic methods of purging tumor cells from stem cell products focus on either removal of specific target cells or positive selection of HPC. mAb are the key component of many purging strategies and are employed both in vitro and in vivo. Cytotoxic cellular therapies are an emerging method of tumor cell eradication. PMID- 10813529 TI - Osmometric and permeability characteristics of human placental/umbilical cord blood CD34+ cells and their application to cryopreservation. AB - The transplantation of placental/cord blood-derived HPC (e.g., CD34+ cells) has become a useful treatment for a broad spectrum of malignant and nonmalignant diseases. The ability to cryopreserve this cell type with high efficiency adds considerable flexibility to cord blood transplantation. The purpose of this study was to develop an understanding of the fundamental cryobiologic factors of these cells, including the osmotic/permeability characteristics, and to use a theoretical approach to optimize freezing procedures. To that end, biophysical parameters, including the osmotically inactive cell volume (Vb), hydraulic conductivity (Lp), and cryoprotectant permeability coefficient (P(CPA)) for DMSO and propylene glycol were measured using a modified Coulter Counter (Coulter Electronics, Inc., Hialeah, FL) at 22 degrees C. In addition, the osmotic tolerance of PCB CD34+ cells was assessed using a colony-forming assay. These experimentally determined parameters were used in a mathematical model to predict optimal cryoprotectant addition and removal procedures. The results demonstrate a Vb of 0.32 x V(iso), an average Lp of 0.17 +/- 0.03 (microm/min/atm +/- SD), and a PCPA of 0.94 +/- 0.004 or 1.0 +/- 0.004 cm/min (x10(-3)) for DMSO or propylene glycol, respectively. No significant difference was determined between the two cryoprotectants used. The osmotic tolerance limits were determined to be 200 and 600 mOsm/kg (1.29 and 0.62 x V(iso), respectively). These results indicate potential benefits of modifications to the widely used method of Rubinstein et al. Proc Natl Acad Sci USA 92:10119-10122, 1995) for cord blood CD34+ cell cryopreservation. As opposed to Rubinstein's method in which DMSO is added to cooled cell suspensions over a 15-min interval, our data indicate that better results may be obtained by introducing and removing the cryoprotectant at ambient temperature over 5 min both to increase viability by avoiding unnecessary risks from osmotic shock and to simplify the protocol. In addition, substitution of propylene glycol for DMSO may be of benefit during the actual freezing and thawing process. PMID- 10813530 TI - Transplantation of gene-modified human bone marrow stromal cells into mouse-human bone chimeras. AB - Transplantation of BM stromal cells engineered to secrete therapeutic factors could represent a treatment for a large array of hematologic disorders. The aim of this study was to evaluate the susceptibility of human BM stromal cell precursors to retroviral gene transfer, then the ability of those to be transplanted in vivo. We have transduced a recombinant retrovirus encoding the mouse CD2 antigen into STRO-1+ cells selected from adult and fetal BM. Gene modified stromal cells were injected intravenously into NOD-SCID mice engrafted previously with pieces of human fetal hematopoietic bone. Using nested PCR, transgenic human cells were detected both in the marrow of human bone grafts and in the BM, liver, and spleen of host mice 7 weeks after grafting. These data indicate that BM stromal progenitor cells are targets for retrovirus-mediated gene transfer and can home to hematopoietic tissues on engraftment through the bloodstream of nonconditioned hosts. PMID- 10813531 TI - In vitro generation of dendritic cells from human blood monocytes in experimental conditions compatible for in vivo cell therapy. AB - DC are professional APC that are promising adjuvants for clinical immunotherapy. Methods to generate in vitro large numbers of functional human DC using either peripheral blood monocytes or CD34+ pluripotent HPC have been developed recently. However, the various steps of their in vitro production for further clinical use need to fit good manufacturing practice (GMP) conditions. Our study focused on setting up such a full procedure, including collection of mononuclear cells (MNC) by apheresis, separation of monocytes by elutriation, and culture of monocytes with GM-CSF + IL-13 + autologous serum (SAuto) in sterile Teflon bags. The procedure was first developed with apheresis products from 7 healthy donors. Its clinical feasibility was then tested on 7 patients with breast cancer. The characteristics of monocyte-derived DC grown with SAuto (or in some instances with a pooled AB serum) were compared with those obtained in the presence of FBS by evaluation of their phenotype, their morphology in confocal microscopy, and their capacity to phagocytize latex particles and to stimulate allogeneic (MLR) or autologous lymphocytes (antigen-presentation tests). The results obtained demonstrate that the experimental conditions we set up are easily applicable in clinical trials and lead to large numbers of well-defined SAuto-derived DC as efficient as those derived with FBS. PMID- 10813532 TI - Suppressive effects of TNF-alpha, TGF-beta1, and chemokines on megakaryocytic colony formation in CD34+ cells derived from umbilical cord blood compared with mobilized peripheral blood and bone marrow. AB - CD34+ cells from human umbilical cord blood (CB) were isolated and investigated for megakaryocytic (MK) colony formation in response to recombinant human (rh) stimulatory and suppressive cytokines and compared with their counterparts in normal BM and G-CSF-mobilized peripheral blood (mPBL). First, we observed that IL 11 by itself at any dosage had no stimulator activity on MK colony formation derived from CD34+ cells in CB, mPBL, and BM. IL-3, steel factor (SLF), or thrombopoietin (Tpo) alone stimulated numbers of colony-forming unit megakaryocyte (CFU-MK) in a dose-dependent fashion. Maximum growth of MK progenitor cells was noted in the presence of a combination of cytokines: IL-11, IL-3, SLF, and Tpo. The frequency of CFU-MK in CB and mPBL was significantly greater than that in BM, and the size of colonies in CB and mPBL was significantly greater than that in BM, and the size of colonies was larger as well. In addition, an increased number of big mixed colonies containing MK were observed in CB and mPBL. In the presence of IL-11, IL-3, SLF, and Tpo, CFU-MK derived from CB, mPBL, and BM was suppressed by tumor necrosis factor-alpha (TNF alpha) and transforming growth factor-beta1 (TGF-beta1). CFU-MK derived from normal BM was inhibited by some chemokines evaluated, whereas CFU-MK derived from CB was suppressed only by platelet factor-4 (PF-4), IFN-inducible protein-10 (IP 10), Exodus-1, Exodus-2, and Exodus-3, but to a lesser degree. In CB, unlike granulocyte-macrophage (CFU-GM), erythroid (BFU-E), high-proliferative potential (HPP-CFC), or multipotential (CFU-GEMM) progenitors, at least a subpopulation of MK progenitors are in S-phase. Therefore, CB MK progenitors respond to the suppressive effects of some members of the chemokine family. Similar results were noted for burst-forming unit-MK (BFU-MK). Our results indicate that CB and mPBL are rich sources of MK progenitors and that MK progenitors in CB are responsive to the suppressive effects of TNF-alpha and TGF-beta1 and some members of the chemokine family. PMID- 10813533 TI - Pharmacokinetics and in vivo effects of a six-base phosphorothioate oligodeoxynucleotide with anticancer and hematopoietic activities in swine. AB - A short phosphorothioate oligodeoxynucleotide telomere mimic with the sequence 5' d(TTAGGG)-3', TAG-6, has been shown to inhibit telomerase activity and have antineoplastic and hematopoietic stimulatory properties. In this study, three immature male domestic swine (weighing approximately 40 kg) were administered 200 mg/m2 of TAG-6 by continuous intravascular infusion at rates of 0.48 +/- 0.07 mg/hr for 14 days to evaluate the pharmacokinetics, toxicity, and tissue distribution. There was considerable variability (both within each animal and across animals) observed in the pharmacokinetic data. The plasma half-life (t1/2 appeared to be short enough that it could be assumed that steady state was attained by at least 96 h after the start of the infusion. The t1/2 estimates for the three pigs were 8.96, 109, and 1.97 h (the long t1/2 for pig 2 may be explained by poor parameter estimation due to the variability). The volume of distribution ranged from 9.80 to 51.8 L (0.3-1.4 L/kg), and plasma clearance estimates ranged from 0.33 to 3.46 L/h (5.5-57.7 ml/min). The average plasma concentrations at steady state were 0.845, 0.933, and 0.178 microg/ml (0.44, 0.49, and 0.093 microM) for the three animals. Nearly 30% of the administered dose was cleared through renal excretion by day 7 postinfusion. The distribution of TAG-6 was primarily to the liver and kidney, but the spleen and thyroid accumulated relatively high concentrations of TAG-6. TAG-6 was metabolized to apparently higher molecular weight products, which were observed in the urine. The size periodicity of these apparently higher molecular weight products was in 6-base intervals, which is consistent with the actions of telomerase. The infusion did not produce significant changes in serum chemistry or circulating blood cells, but a decrease in colony-forming unit-granulocyte-monocyte (CFU-GM) colony formation from BM was observed. These data suggest that TAG-6 may be a very specific pharmacophore. PMID- 10813534 TI - Protection of hematopoietic progenitor cells by nitric oxide. AB - The free radical gas nitric oxide (NO) has been reported to trigger apoptosis of a variety of cell types, but there have also been contradictory reports of an inhibitory effect of NO on this form of programmed cell death. We hypothesized that NO may inhibit apoptosis of hematopoietic cells. In this study, therefore, hematopoietic cells obtained from umbilical cord blood (CB) were incubated with NO, and NO inhibited apoptosis of hematopoietic cells in umbilical CB, although it did not affect the number of mononuclear cells (MNC). Inhibition of apoptosis of hematopoietic cells is important for their use in transplantation. Our results suggest that it might be possible to prevent the loss of hematopoietic cells using NO. PMID- 10813535 TI - Automated immature reticulocyte counts are early markers of engraftment following autologous PBSC transplantation in patients with lymphoma. AB - The traditional indicators of engraftment following PBSC transplantation (PBSCT) are the rising total WBC count and ANC. Reticulocytes may be an earlier indicator, since as reticulocytes mature, there is a gradual loss of cellular RNA, which can be measured using methylene blue and light scatter with an Abbott CD 3500 automated counter (Abbott Laboratories, Maidenhead, U.K.). Reticulocytes can be divided into three fluorescence ratios depending on the amount of light scatter generated, high, medium, and low. The most immature are the high fluorescence reticulocytes (HFR). Standard engraftment parameters together with HFR were measured in a homogeneous group of 25 patients with lymphoma after PBSCT using a standard conditioning protocol. An ANC of 0.5 x 10(9)/L was achieved after a median of 10 days (mean 11.2 days, range 9-22). The recovery of the HFR to 2% of the total reticulocytes was significantly shorter, with a median of 8 days (mean 7.5 days, range 6-10) (p < 0.0001). The values of HFR to 2% preceded the ANC of 0.5 x 10(9)/L in 24 of the 25 patients by a median of 3 days (mean 3.8 days, range 2-12 days). On this basis, it can be determined that in 96% of cases, engraftment was indicated earlier by HFR measurement. The HFR to 2% even preceded the ANC of 0.1 x 10(9)/L in 23 of the 25 patients, showing that engraftment was indicated earlier in 92% of patients. Immature reticulocytes appearing in peripheral blood can be reliably measured by automated cytometers, and HFR can, therefore, be used as an earlier indicator of engraftment following PBSCT. This information provides the opportunity for earlier cessation of antibiotics and growth factors and could lead to earlier discharge from hospital, with cost savings. PMID- 10813536 TI - Development of a competitive PCR method for in vitro and in vivo quantification of herpes simplex virus thymidine kinase and neomycin resistance-expressing cells used in a clinical trial. AB - The aim of this study was to set up a sensitive and specific method to quantify the number of gene-modified cells in a gene therapy clinical trial currently underway at our institution. This trial involves the use of retrovirally transduced allogeneic T cells expressing the herpes simplex-1 thymidine kinase (HSV-TK) and neomycin-phosphotransferase (NeoR) resistance gene. Quantification by competitive PCR was performed, with two homologous internal standards (deltaTK, deltaNeoR), 30 bp shorter than the target sequences (TK, NeoR), coupled to fluorescent laser-based detection. Assessment of the amplification systems procedures was carried out for each sequence. The 30-bp deletion did not affect the amplification efficiency significantly. Determination of the plateau phase of both amplified sequences demonstrated that each sample must be quantified during the predetermined exponential phase. Finally, a blinded study of a transduced cell dilutions panel validated the overall methodology. The competitive PCR was applied to quantification of the retroviral transduction process by quantifying the NeoR gene in transduced PBMC samples (prior to G418 selection) from 18 donors in our clinical trial. A mean transduction efficiency of 9.78% +/- 1.37% was observed. We also quantified TK-expressing donor transgenic T cells in a murine GvHD model. Results demonstrated on initial expansion of donor HSV-TK- expression T cells as well as a significant ganciclovir (GCV)-induced decrease correlated with the number of circulating gene-modified T cells. Therefore, we have developed an efficient gene quantification tool that should be useful for in vivo monitoring of gene-modified cells. PMID- 10813537 TI - A comparison of macrophage colony-stimulating factor (M-CSF) gene expression in primary and immortalized endothelial cells. AB - M-CSF is produced by a wide variety of cell types, including EC, fibroblasts, and monocyte/macrophages, where it functions as a survival factor and a chemotactic agent for monocytes. An early event in the development of atherosclerosis is the infiltration of monocytes into the artery wall. Local expression of M-CSF by EC lining the blood vessels is thought to promote the growth and survival of lesional monocytes and macrophages, thus enhancing lesion development and disease progression. Primary cultures of EC are difficult to maintain for long periods of time, which complicates their use for biochemical and molecular analysis. As a step toward identifying a representative endothelial-like cell line, serum dependent and IL-1-dependent changes in M-CSF gene expression in two endothelial like cell lines were compared to that detected in primary EC cultures. The data presented here demonstrate that the two endothelial-like cell lines, like primary cultures of EC, express the M-CSF gene under basal conditions. In both types of cell cultures, IL-1alpha stimulation increased M-CSF mRNA levels 2-7-fold, whereas serum stimulation elicited a more modest effect (2-3-fold increase). The IL-1alpha-induced change in M-CSF gene expression is mediated at the transcriptional level, and M-CSF promoter activity is, in part, dependent on the activity of the NF-kappaB-inducing kinase. Collectively, our results demonstrate that either endothelial-like cell line would be a representative model in which endothelial-specific changes in M-CSF gene expression could be identified. PMID- 10813538 TI - The clinical and biologic significance of abnormal lipid profiles in patients with myelodysplastic syndromes. AB - Serum lipid profiles were obtained in 108 patients with myelodysplastic syndrome (MDS) and compared to 28 healthy volunteers. Serum cholesterol and low-density and high-density lipoproteins (LDL and HDL) were found to be significantly lower in MDS patients than in normals (p = 0.0001, 0.0038 and 0.037, respectively). This difference was significant for all MDS categories. Serum cholesterol and HDL were negatively related to biopsy cellularity (p = 0.001 and 0.0001, respectively), and serum triglycerides were negatively related to labeling index (p = 0.0003). No differences were noted in the lipid profiles of MDS patients with normal versus abnormal karyotypes. However, low-risk MDS patients with abnormal karyotypes had significantly lower triglyceride levels compared with the high-risk patients (p = 0.027), as did low-risk patients with normal cytogenetics (p = 0.015). Serum HDL levels were significantly higher for the low-risk group with normal cytogenetics as well (p = 0.003). We conclude that serum cholesterol, LDL, and HDL are significantly reduced in MDS patients, probably indicating excessive intracellular lipid biosynthesis in the expanding clone. These relatively simple measurements could serve as important prognostic markers and reliable indicators of disease activity in individual patients. Prospective studies to determine their utility as independent variables that guide the need for active therapeutic intervention are warranted. PMID- 10813539 TI - Prevalence of MLR blocking antibodies before and after immunotherapy. AB - Immunologically specific blocking factors (BF) are believed to play a protective role in the maintenance of pregnancy. We compared the levels of BF in normal pregnant women and in patients with recurrent spontaneous abortion (RSA) and observed that the two groups differ significantly from one another. MLC inhibitory activity was present throughout normal pregnancy and was found to be specific to the husband's HLA antigens. Immunotherapy was performed in 28 recurrent aborters using husband's lymphocytes, after which levels of blocking antibodies increased in 23 patients. The difference in the label of MLR BF in the preimmunotherapy and postimmunotherapy groups was statistically significant and was associated with successful pregnancy outcome in 82.15% of cases. The appearance of the MLR BF and its continuation in a successful pregnancy might represent an appropriate immune state that may be playing an important role in maintenance of pregnancy. PMID- 10813540 TI - Molecular assessment of post-BMT chimerism using various biologic specimens and automated DNA sizing technology. AB - Highly polymorphic microsatellite markers provide useful genetic markers for detection of complete or mixed chimerism in patients after allogeneic BMT (allo BMT). We report application of automated DNA sizing technology for detection of post-BMT chimerism using fresh peripheral blood, BM, or archival blood smears and various DNA isolation techniques. Donors' and recipients' DNA was amplified with fluorescent PCR primers specific for short tandem repeat (STR) marker loci: FGA, VWA, TH01, F13A1, D21S11. Chimerism was assessed in 14 recipients after allo-BMT. A complete chimerism was detected in 10 patients, in 3 patients we observed fluctuations of chimerism status, and mixed chimerism was assessed in 1 patient. We show that DNA from different types of biologic specimens (whole peripheral blood, BM suspension, archival blood smears), prepared according to the various isolation techniques (salting-out method, phenol chloroform extraction, Chelex procedure) and amplified with fluorescent PCR primers for microsatellite markers, enable identification of chimerism status following allo-BMT in children. PMID- 10813541 TI - A randomized trial of leukapheresis volumes, 7 L versus 10 L: an assessment of efficacy and patient tolerance. AB - High-dose chemotherapy followed by autologous PBSC transplantation (PBSCT) has become an accepted form of therapy for a number of malignant hematologic diseases. The optimal method for the collection of PBSC is yet to be defined. Large-volume leukapheresis may be able to collect adequate numbers of PBSC with the patient undergoing fewer procedures. We routinely process 7 L of blood per leukapheresis. Hence, we elected to assess whether a modest increase in the blood volume processed would, on average, decrease the number of leukaphereses each patient needed to undergo to collect > or =2 x 10(6) CD34+ cells/kg body weight. Sixty patients were randomized to undergo 7 L leukaphereses (n = 31 patients; 87 leukaphereses) or 10 L leukaphereses (n = 29 patients; 81 leukaphereses). The median number of leukaphereses required per patient to collect the target number of CD34+ cells was two (range one to five) for both groups (p = 0.83). The median number of nucleated cells collected per patient was greater for the 10 L group (8.2 x 10(8)/kg versus 5.3 x 10(8)/kg, p = 0.005), as was the median number of mononuclear cells (MNC) (4.7 x 10(8)/kg versus 3.6 x 10(8)/kg, p = 0.0001), whereas there was no statistical difference between the groups for the median number of CD34+ cells collected per patient (3.2 x 10(6)/kg versus 3.7 x 10(6)/kg, p = 0.98). Therefore, over the 18-month period of this trial, the use of a 10 L leukapheresis volume did not decrease the number of leukaphereses performed compared with a 7 L leukapheresis volume. PMID- 10813542 TI - Competitive cytokeratin 19 RT-PCR for quantification of breast cancer cells in blood cell suspensions. AB - Detection of residual tumor cells in BM and PBPC products has been correlated with worse outcome of breast cancer patients. Still, there is a considerable demand for studies investigating the influence of the actual tumor cell number on prognosis, as quantification routinely has been cumbersome and time consuming and, thus, was evaded. We developed and evaluated a competitive RT-PCR-ELISA assay for cytokeratin 19 (CK19) with standard curve quantification that allows quantification of multiple samples within a working day; mRNA isolation, RT-PCR reaction, and automated ELISA detection were carried out using commercial kits. Results were expressed as OD420nm ratios of CK19 and an internal competitor. Values were then converted into tumor cell numbers using a standard curve of MCF 7 tumor cells. The assay had high specificity because of primers and capture probes with great heterogeneity to both published pseudogenes, which was confirmed by BLAST sequence alignment. We achieved a sensitivity of detecting 1 tumor cell per 10(6) mononuclear cells (MNC). Between-batch precision (n = 8) for quantification was consistent and reasonable, with a coefficient of variation around 25%. Therefore, this assay should be suitable and sufficient for routine quantification of tumor cell numbers in BM or PBPC samples. PMID- 10813543 TI - High steady-state plasma levels of flt3-ligand in the peripheral blood is a good predictor for poor mobilization of CD34+ PBSC in patients undergoing high-dose chemotherapy and stem cell rescue. AB - flt3-ligand (flt3-L) is a very effective mobilizer of hematopoietic stem cells (HSC) and is capable of inducing multilineage hematopoietic cell differentiation both in vivo and in vitro. We measured, by ELISA, the plasma peripheral blood flt3-L concentrations in 28 non-Hodgkin's lymphoma (NHL) patients before and after mobilization with three different mobilization regimens, including priming with cyclophosphamide (CY) plus G-CSF, CY plus GM-CSF, and CY plus GM-CSF (8 days) followed by G-CSF. We also determined the levels of flt3-L in the peripheral blood during four apheresis collections and 6 months after transplantation. The steady-state level of flt3-L in NHL patients (n = 18) who mobilized > or =2 x 10(6) CD34+ cells/kg in four apheresis collections was 34 +/- 4 pg/ml and was similar to the levels observed in 10 normal controls (27 +/- 7, p = 0.1) regardless of the mobilization protocol used. In contrast, patients who failed to mobilize a total of >0.4 x 10(6) CD34+ cells/kg in two consecutive apheresis collections (n = 10) had flt3-L levels of 106 +/- 11 pg/ml, significantly higher (p = 0.006) than that of the good mobilizers group, regardless of the mobilization protocol used. Similar results were observed in 29 multiple myeloma (MM) patients. A mean of 23.8 +/- 7.9 pg/ml and 450 +/- 85 pg/ml flt3-L was obtained in the good mobilizers (n = 24) and the nonmobilizers (n = 5) groups of patients, respectively. Statistical analysis revealed a significant difference (p = 0.0006) between the two groups of MM patients, but no correlation was observed between the levels of flt3-L and CD34+ cell/microl, in mobilized peripheral blood. Our results also suggest that measurement of plasma levels of flt3-L before mobilization can be clinically useful to predict for patients with poor mobilization outcome. PMID- 10813544 TI - Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals. PMID- 10813545 TI - Skin permeation of testosterone and its ester derivatives in rats. AB - To establish the optimum conditions for improving the transdermal delivery of testosterone, we studied the relationship between the lipophilicity of testosterone ester derivatives and the rat skin permeation rate of testosterone. We performed a rat skin permeation study of testosterone and its commercially available ester derivatives, testosterone hemisuccinate, testosterone propionate and testosterone-17beta-cypionate, using an ethanol/water co-solvent system. The aqueous solubility and rat skin permeation rate of each drug, saturated in various compositions of an ethanol/water system, was determined at 37 degrees C. The aqueous solubility of testosterone and its ester derivatives increased exponentially as the volume fraction of ethanol increased up to 100% (v/v). The stability of testosterone propionate in both the skin homogenate and the extract was investigated to observe the enzymatic degradation during the skin permeation process. Testosterone propionate was found to be stable in the isotonic buffer solution and in the epidermis-side extract for 10h at 37 degrees C. However, in the skin homogenate and the dermis-side extract testosterone propionate rapidly degraded producing testosterone, implying that testosterone propionate rapidly degraded to testosterone during the skin permeation process. The steady-state permeation rates of testosterone in the ethanol/water systems increased exponentially as the volume fraction of ethanol increased, reaching the maximum value (2.69+/-0.69 microg cm(-2)h(-1)) at 70% (v/v) ethanol in water, and then decreasing with further increases in the ethanol volume fraction. However, in the skin permeation study with testosterone esters saturated in 70% (v/v) ethanol in water system, testosterone esters were hardly detected in the receptor solution, probably due to the rapid degradation to testosterone during the skin permeation process. Moreover, a parabolic relationship was observed between the permeation rate of testosterone and the log P values of ester derivatives. Maximum flux was achieved at a log P value of around 3 which corresponded to that of testosterone (log P = 3.4). The results showed that the skin permeation rate of testosterone and its ester derivatives was maximized when these compounds were saturated in a 70% ethanolic solution. It was also found that a log P value of around 3 is suitable for the skin permeation of testosterone related compounds. PMID- 10813546 TI - The encapsulation of bleomycin within chitosan based polymeric vesicles does not alter its biodistribution. AB - Polymeric vesicles have recently been developed from an amphiphilic chitosan derivative--palmitoyl glycol chitosan. Their potential as a drug delivery system was evaluated using the anti-cancer compound bleomycin as a model drug. Palmitoyl glycol chitosan (GCP41) was synthesised by conjugation of palmitoyl groups to glycol chitosan. Bleomycin-containing vesicles (669 nm diameter) were prepared from a mixture of GCP41 and cholesterol by remote loading. The vesicles were imaged by freeze-fracture electron microscopy and their in-vitro stability tested. Incubation of the larger vesicles with plasma in-vitro led to a reduction of mean size by 49%, a reaction not seen with control sorbitan monostearate niosomes (215 nm in size). They also showed a higher initial drug release (1 h), but GCP41 and sorbitan monostearate vesicles retained 62% and 63% of the encapsulated drug after 24h, respectively. The biodistribution of smaller vesicles (290 nm) prepared by extrusion through a 200-nm filter was also studied in male Balb/c mice. Encapsulation of bleomycin into polymeric vesicles did not significantly alter the pharmacokinetics of biodistribution of bleomycin in male Balb/c mice although plasma and kidney levels were slightly increased. It is concluded that the extruded GCP41 vesicles break down in plasma in-vivo and hence are unlikely to offer any therapeutic advantage over the free drug. PMID- 10813547 TI - Enhancement of ursodeoxycholic acid bioavailability by cross-linked sodium carboxymethyl cellulose. AB - The bioavailability of ursodeoxycholic acid from a new formulation based on drug loaded cross-linked sodium carboxymethyl cellulose was studied in man. The plasma levels of ursodeoxycholic acid were determined by gas chromatography-mass spectrometry after derivatization and sample purification by solid-phase extraction. Capsules containing the drug/polymer system were prepared and compared with conventional commercial ursodeoxycholic acid capsules after single oral administration using a randomized crossover experimental design. Although the drug/polymer system improved the in-vitro dissolution rate of ursodeoxycholic acid in simulated intestinal fluid, statistical evaluation of the area under the plasma concentration curves indicated no significant difference in the extent of bioavailability between the two formulations (14.93+/-4.43 vs 14.95+/-5.79 microM h; P > 0.2). However, following the administration of the ursodeoxycholic acid/cross-linked sodium carboxymethyl cellulose system with an enteric-coated capsule, the mean area under the plasma concentration curve (27.60+/-10.11 microM h) was significantly higher than that obtained after treatment with the commercially available ursodeoxycholic acid capsule (16.24+/-8-38 microM h; P < 0.05). We concluded that improved intestinal absorption of the drug was obtained with enteric-coated capsules filled with the ursodeoxycholic acid/polymer system. Moreover, the simplicity of the preparation and the non-toxicity of the polymer used as the carrier represented additional advantages of this dosage form. PMID- 10813548 TI - Chronic continuous cocaine infusion in rats: effect on urine cocaine, ecgonine methylester and benzoylecgonine concentrations and bolus-dose cocaine pharmacokinetics. AB - The aim of this study was to determine the effect of chronic cocaine infusion on urine cocaine, ecgonine methylester and benzoylecgonine concentrations to establish if they varied with dose and duration of cocaine administration. Male rats were continuously infused with cocaine at either 6 or 18 mg kg(-1) daily for 13 days. Three urine samples taken over the course of the infusion period showed that cocaine, ecgonine methylester and benzoylecgonine concentrations varied with the dose administered and the duration of administration. Cocaine, ecgonine methylester and benzoylecgonine concentrations were 2-3 times greater in the high dose group than the low-dose group at each sampling time point. These decreased, respectively, from 7.0+/-1.1, 26.7+/-4.5 and 29.5+/-5.4 microg mL(-1) to 2.5+/ 0.5, 10.5+/-1.8 and 11.8+/-1.5 microg mL(-1) in the high-dose group and from 1.0+/-0-2, 7.8+/-1.5 and 6.3+/-0.1 microg mL(-1) to 0.5+/-0.1, 4.0+/-0.6 and 3.1+/-0.4 microg mL(-1) in the low-dose group (P < 0.05) over the infusion period. We also studied the pharmacokinetic and metabolic profile of an intravenous bolus dose of 2.5 mg kg(-1) cocaine hydrochloride after a similar cocaine infusion in rats. Cocaine pharmacokinetics and the profile of ecgonine methylester, benzoylecgonine and norcocaine were no different from rats chronically infused with saline for the same period. Altered cocaine metabolism could not explain the effect of the duration of cocaine infusion on altered metabolite concentrations in urine. Ecgonine methylester/benzoylecgonine urine concentration ratios did not alter with duration of infusion (1.2+/-0.2 and 1.1+/ 0.2 in the high-dose group at the first and last time point) and were not affected by the dose of cocaine (1.3+/-0.6 and 1.2+/-0.1 at corresponding times in the low-dose group (P > 0.05)). We conclude that chronic cocaine infusion does not alter cocaine metabolism. This was not reflected by absolute cocaine metabolite urine concentrations, which varied with time, but was represented by urine ecgonine methyl ester/benzoylecgonine concentration ratios. PMID- 10813549 TI - Effects of ascorbic acid on interactions between ciprofloxacin and ferrous sulphate, sodium ferrous citrate or ferric pyrophosphate, in mice. AB - The absorption of ciprofloxacin has been reported to be impaired by concomitant administration of ferrous sulphate. The effects of sodium ferrous citrate and ferric pyrophosphate, which have been used as extensively as ferrous sulphate, on the absorption of ciprofloxacin were compared with that of ferrous sulphate. The effects of ascorbic acid on the interactions between ciprofloxacin and each iron compound were studied in mice. Mice were treated orally with ciprofloxacin (50 mg kg(-1)) alone, the iron compound (ferrous sulphate, sodium ferrous citrate or ferric pyrophosphate; 50 mg elemental iron kg(-1)) alone, ciprofloxacin with each iron compound or ciprofloxacin in combination with each iron compound and ascorbic acid (250 mg kg(-1)). The maximum serum concentration of ciprofloxacin was significantly (P < 0.01) reduced from 1.15+/-0.11 microg mL(-1) (ciprofloxacin alone) to 0.17+/-0.01, 0.27+/-0.01 or 0.28+/-0.02 microg mL(-1), respectively, when ferrous sulphate, sodium ferrous citrate or ferric pyrophosphate was administered along with ciprofloxacin. The addition of ascorbic acid did not affect the inhibitory effects of each iron compound on the absorption of ciprofloxacin. Ciprofloxacin did not affect the variation of serum iron levels after administration of each iron compound. The addition of ascorbic acid significantly (P < 0.01) enhanced the increase in serum iron concentration after administration of sodium ferrous citrate, showing an increase from 270+/-6 microg dL(-1) to 463+/-11 microg dL(-1) compared with an increase from 248+/-8 microg dL(-1) to 394+/-18 microg dL(-1) after administration of sodium ferrous citrate alone. Ascorbic acid also caused a significant (P < 0.01) increase in serum iron concentration from 261+/-16 microg dL(-1) to 360+/-12 microg dL(-1) after administration of ferric pyrophosphate, although it did not affect the levels after ferrous sulphate administration. The results suggest that sodium ferrous citrate and ferric pyrophosphate should not be administered with ciprofloxacin (as for ferrous sulphate) and that sodium ferrous citrate is converted to the ferric form more easily than ferrous sulphate. This difference in convertibility might contribute to a clinical difference between sodium ferrous citrate and ferrous sulphate. PMID- 10813550 TI - Effects of trimebutine maleate on delayed rectifier K+ currents in guinea-pig ventricular myocytes. AB - The effects of trimebutine maleate, a drug commonly used to regulate motility in the gastrointestinal tract, on the delayed rectifier K+ current (I(K)) were evaluated in guinea-pig ventricular myocytes to determine whether the drug has a proarrhythmic effect through blockade of I(K). Trimebutine decreased I(K) in a concentration-dependent manner. To investigate the effects of trimebutine on two components of I(K) (I(Kr) and I(Ks); rapidly activated and slowly activated components, respectively), we performed the envelope-of-tails test. Trimebutine sensitive I(K) was determined by digital subtraction of I(K) during exposure to trimebutine from control I(K) for each duration of the test pulse over the range 50 ms-2 s. The ratio of deltaI(K,tail)/deltaI(K) plotted against pulse duration for trimebutine-sensitive I(K) gradually decreased to a steady-state value as the duration of the test pulse was lengthened. This finding suggested a weak inhibitory effect of trimebutine on both I(Kr) and I(Ks). The effects of trimebutine on the inward rectifier K+ current (I(K1)) responsible for the resting potential and final repolarization phase of the action potential were investigated by applying voltage clamp ramps over a broad range of potentials. No significant effects were observed at 10 or 100 microM. We next investigated the effects of the drug on the L-type Ca2+ current (I(Ca)). Significant inhibition of I(Ca) was observed at trimebutine concentrations greater than 10 microM. These results suggested that trimebutine maleate has weak inhibitory effects on I(Kr), I(Ks) and I(Ca) at concentrations much higher than those in clinical use. PMID- 10813551 TI - Endothelium-dependent sensory non-adrenergic non-cholinergic vasodilatation in rat thoracic aorta: involvement of ATP and a role for NO. AB - The involvement of non-adrenergic non-cholinergic (NANC) transmitters, such as adenosine 5'-triphosphate (ATP) and nitric oxide (NO), in the neurogenic relaxation of rat thoracic aorta was investigated in vessel segments suspended for isometric tension recording by polygraph. Responses to electrical field stimulation (EFS) and exogenous vasodilator were investigated in vessels precontracted with 5-hydroxytryptamine. EFS (100 V, 2-16 Hz, for 10 s at 3-min intervals), in the presence of guanethidine (10 microM) and atropine (10 microM) produced frequency-dependent relaxations. Pretreatment with tetrodotoxin (1 microM) markedly reduced the relaxation and desensitization with capsaicin (10 microM) significantly inhibited the relaxation. Exogenously added ATP caused concentration-dependent relaxations. Mechanical removal of the endothelium significantly inhibited EFS- and ATP-induced relaxation by 30+/-3% and 37+/-2%, respectively. Pretreatment with a P1-purinoceptor antagonist, 8 phenyltheophylline (10 microM) or P2X-purinoceptor antagonist, Evans blue (10 microM) did not influence the relaxations to EFS and exogenously added ATP. In contrast, the P2Y-purinoceptor antagonist, basilen blue (100 microM) markedly reduced the relaxations to EFS by 52+/-4% in the endothelium-intact preparations. However, in the endothelium-denuded preparations and capsaicin-pretreated preparations, basilen blue did not change relaxations elicited by EFS. The NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM) also significantly inhibited the relaxations to EFS and ATP by 40+/-6% and 30+/-2%, respectively, in the endothelium-intact preparations but had no effect on the relaxations in the endothelium-denuded preparations or capsaicin-pretreated preparations. In addition, the EFS-induced relaxations were also inhibited 43+/ 7% by pretreatment with 1H-[1,2,4]-oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ; 1 microM), soluble guanylate cyclase inhibitor. This study suggests that the NANC nerve system is present in the thoracic aorta of rat, mediating vasodilatation by sensory nerves. ATP, as a neurotransmitter released from sensory nerves, activates P2Y-purinoceptors located on the endothelium and stimulates the NO/cyclic GMP pathway, resulting in vasodilatation. PMID- 10813553 TI - Fluvastatin, a new inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, resists hydroxyl radical generation in the rat myocardium. AB - The aim of this study was to determine whether fluvastatin, an inhibitor of low density lipoprotein (LDL) oxidation, can resist Cu (II)-induced hydroxyl radical generation (*OH) in the extracellular fluid of rat myocardium. Rats were anaesthetized and sodium salicylate in Ringer's solution (0.5 nmol microL(-1) min(-1)) was infused through a microdialysis probe to detect the generation of *OH as reflected by the non-enzymatic formation of 2,3-dihydroxybenzoic acid (2,3 DHBA) in the myocardium. When Cu (II) (0, 10, 25 or 50 microM) was administered to 1 mM tyramine-pretreated rats, a marked elevation in the levels of 2,3-DHBA was found, indicating a positive linear correlation between Cu (II) and the increase in *OH formation trapped as 2,3-DHBA in the dialysate (r2 = 0.977). In the presence of fluvastatin (100 microM), a marked decrease in the levels of 2,3 DHBA was found. Corresponding experiments performed with iron (II) (0, 10, 25 or 50 microM), showed a marked elevation in the levels of 2,3-DHBA, indicating a positive linear correlation between iron (II) and the increase in *OH formation trapped as 2,3-DHBA in the dialysate (r2 = 0.986). However, in the presence of fluvastatin (100 microM) a small decrease in the level of 2,3-DHBA was found. The results show that iron (II) against LDL oxidation may be insensitive compared with Cu (II). Cu (II)-induced *OH formation may be reduced by inhibiting LDL with fluvastatin. PMID- 10813552 TI - Differential potency of beclomethasone esters in-vitro on human T-lymphocyte cytokine production and osteoblast activity. AB - Beclomethasone dipropionate is an inhaled corticosteroid, used for the treatment of asthma. It is metabolised to 17-beclomethasone monopropionate, which has greater affinity for corticosteroid receptors than the parent compound, and to beclomethasone. We investigated the potency of beclomethasone dipropionate, 17 beclomethasone monopropionate and beclomethasone (compared with dexamethasone as a reference steroid) in two different human cell types, peripheral blood mononuclear cells and osteoblasts. We found that beclomethasone dipropionate, 17 beclomethasone monopropionate (EC50 10(-14) M) and beclomethasone (EC50 approx. 10(-12) M) were much more potent than dexamethasone (EC50 10(-8) M) in inhibiting interleukin-5 production by peripheral blood mononuclear cells. In contrast, beclomethasone dipropionate, 17-beclomethasone monopropionate and beclomethasone were equipotent with dexamethasone (EC50 range 0.3-1.2 x 10(-9) M) in affecting several functional assays of osteoblasts (e.g. alkaline phosphatase activity and osteocalcin synthesis). These results show that the relative bioactivities of corticosteroids vary between different human cell types, and that affinities observed in receptor binding assays are not necessarily predictive of the bioactivity in cell populations, such as peripheral blood mononuclear cells and osteoblasts, which are putatively relevant to efficacy and side effects respectively. PMID- 10813554 TI - The role of nitric oxide and prostaglandin E2 on the hyperalgesia induced by excitatory amino acids in rats. AB - The present study was designed to investigate the role of nitric oxide (NO), N methyl-D-aspartate (NMDA) receptor and prostaglandins on hyperalgesia induced in rats by excitatory amino acids and the possibility that prostaglandins may act as the retrograde messenger in the spinal cord like NO. Nomega-nitro-L-arginine methyl ester (L-NAME; 500 microg/paw, intraplantarly (i.pl.)), MK-801 (10 microg/paw, i.pl.) or indomethacin (300 microg/paw, i.pl.) reduced the duration of phase 2 of the biting/licking and scratching (B/L + S) response induced by formalin injection from 255.6+/-16.7 s to 155.6+/-16.9, 172.25+/-33.3 or 205.6+/ 16.7 s, respectively. L-NAME (0.3 mg, i.th.), MK-801 (8 microg, i.th.) or indomethacin (20 microg, i.th) reduced the duration of phase 2 of the B/L + S response induced by saline injection from 288.5+/-7.7s to 207.7+/-19.2, 184.6+/ 7.7 or 1923+/-38.5 s, respectively. L-NAME or indomethacin injected into the spinal cord of the rat significantly reduced the hyperalgesia induced by NMDA (1 microg, i.th.) from 43.8+/-4.6% to 12.3+/-3.1 and 19.2+/-2.3%, respectively. It is assumed that NO produced by excitatory amino acids may increase prostaglandin production by cyclooxygenase activation. L-NAME, MK-801 or indomethacin injected into the rat spinal cord significantly reduced the hyperalgesia induced by prostaglandin E2 (PGE2, 25 ng, i.th.) in the tail-flick test from 40.6+/-3.5% to 18.2+/-3.2, 18.8+/-1.8 or 17.6+/-4.1%, respectively, but had little effect on hyperalgesia in the paw pressure test (except for indomethacin). In conclusion, NO and PGE2 affect the hyperalgesia induced by excitatory amino acids. It is suggested that PGE2, like NO, may act as a retrograde messenger in the spinal cord. PMID- 10813555 TI - Protective effect of curcumin in rat liver injury induced by carbon tetrachloride. AB - This study was carried out to investigate the protective effects of curcumin on acute or subacute carbon tetrachloride-induced liver damage in rats. Acute hepatotoxicity was induced by intraperitoneal injection of carbon tetrachloride after 4 consecutive days of curcumin treatment. Subacute hepatotoxicity was induced by oral administration of carbon tetrachloride twice a week during 4 weeks of curcumin treatment. In rats with acute liver injury, curcumin (100 and 200 mg kg(-1)) lowered the activity of serum alanine aminotransferase to 52-53% (P < 0.05) and aspartate aminotransferase to about 62% (P < 0.05) those of control rats. In rats with subacute liver injury, curcumin (100 mg kg(-1)) lowered the activity of serum alanine aminotransferase to 34% (P < 0.01) and alkaline phosphatase to 53% (P < 0.05) of control rats. The liver hydroxyproline content in the curcumin (100 mg kg(-1))-treated group was reduced to 48% of the carbon tetrachloride control group (P < 0.01). Malondialdehyde levels in curcumin (100 mg kg(-1)) treated rat liver was decreased to 67% of the control rat liver (P < 0.01) in subacute injury. It was concluded that curcumin improved both acute and subacute liver injury induced by carbon tetrachloride in rats. PMID- 10813556 TI - SM-20220, a potent Na+/H+ exchange inhibitor, improves consciousness recovery and neurological outcome following transient cerebral ischaemia in gerbils. AB - We studied the cerebroprotective effect of SM-20220 (N-(aminoiminomethyl)-1 methyl-1H-indole-2-carboxamide methanesulphonate), a newly synthesized Na+/H+ exchanger (NHE) inhibitor, in Mongolian gerbil global ischaemia. Transient cerebral ischaemia was induced by clipping both common carotid arteries for 30 min followed by 24h reperfusion. Intravenous administration of SM-20220 (0.3 or 1.0 mg kg(-1)) immediately after reperfusion significantly shortened the consciousness recovery time (P < 0.01). SM-20220 also improved the neurological outcome (McGraw's scale) after reperfusion. At the dose of 1.0 mg kg(-1), the mortality rate was significantly reduced at 24 h after reperfusion (P < 0.01). This study shows that NHE is involved in the aggravation of cerebral function, represented by consciousness recovery, and neurological outcome following transient forebrain ischaemia, and that its inhibitor may exert protective effects on post-ischaemic brain damage. PMID- 10813557 TI - Estimation of individual sennosides in plant materials and marketed formulations by an HPTLC method. AB - Senna is a well-known drug, used in the Ayurvedic and Allopathic systems of medicine, and is a treatment for constipation. The purgative action of senna and its formulations is due to the presence of sennosides A and B. An HPTLC method has been developed for the determination of individual sennosides (A, B, C, D) without any derivatization in marketed formulations (three tablet formulations, two granule formulations and one liquid formulation) and plant materials (senna leaf and pod). The methanolic solution of a sample was applied on a pre-coated silica gel G60 F254 TLC plate (E. Merck.) and was developed using n-propanol : ethyl acetate : water : glacial acetic acid (3 : 3 : 2 : 0.1 v/v) as the mobile phase. The relative band speeds (Rf values) obtained were 0.35, 0.25, 0.61, 0.46 for sennosides A, B, C and D, respectively. The densitometric response was monitored at 366nm. Calibration curves were found to be linear in the concentration ranges 193-1356, 402-2817, 71-497 and 132-927 ng per spot for sennosides A, B, C, and D, respectively. The correlation coefficients were found to be 0.9978, 0.9987, 0.9939 and 0.9983 respectively for sennosides A, B, C and D. The result obtained with the HPTLC method for total sennoside content was compared with the results using the pharmacopoeial methods (spectrophotometric (British Pharmacopoeia) and spectrofluorimetric (United States Pharmacopeia) using the 'F' test). The results revealed no significant difference in the three different methods for estimation of total sennoside. The proposed HPTLC method was found to be simple, specific, precise, accurate and rapid. It can be used for routine quality control of sennosides or senna-containing formulations for individual sennosides. PMID- 10813558 TI - Identification of kaempferol as a monoamine oxidase inhibitor and potential Neuroprotectant in extracts of Ginkgo biloba leaves. AB - The effects of Ginkgo biloba leaf extract on rat brain or livermonoamine oxidase (MAO)-A and -B activity, biogenic amine concentration in nervous tissue, N-methyl D-aspartate (NMDA)- and N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) induced neurotoxicity and antioxidant activity was investigated to determine the effects of the extract on monoamine catabolism and neuroprotection. Ginkgo biloba leaf extract was shown to produce in-vitro inhibition of rat brain MAO-A and -B. The Ginkgo biloba extract was chromatographed on a reverse-phase HPLC system and two of the components isolated were shown to be MAO inhibitors (MAOIs). These MAOIs were identified by high-resolution mass spectrometry as kaempferol and isorhamnetin. Pure kaempferol and a number of related flavonoids were examined as MAOIs in-vitro. Kaempferol, apigenin and chrysin proved to be potent MAOIs, but produced more pronounced inhibition of MAO-A than MAO-B. IC50 (50% inhibition concentration) values for the ability of these three flavones to inhibit MAO-A were 7 x 10(-7), 1 x 10(-6) and 2 x 10(-6) M, respectively. Ginkgo biloba leaf extract and kaempferol were found to have no effect ex-vivo on rat or mouse brain MAO or on concentrations of dopamine, noradrenaline, 5-hydroxytryptamine and 5 hydroxyindoleacetic acid. Kaempferol was shown to protect against NMDA-induced neuronal toxicity in-vitro in rat cortical cultures, but did not prevent DSP-4 induced noradrenergic neurotoxicity in an in-vivo model. Both Ginkgo biloba extract and kaempferol were demonstrated to be antioxidants in a lipid peroxidation assay. This data indicates that the MAO-inhibiting activity of Ginkgo biloba extract is primarily due to the presence of kaempferol. Ginkgo biloba extract has properties indicative of potential neuroprotective ability. PMID- 10813559 TI - Protection by Osbeckia aspera against carbon tetrachloride-mediated alterations in microsomal drug metabolizing enzyme activity. AB - Previous investigations have confirmed the protective effect of Osbeckia aspera leaf extract on carbon tetrachloride-mediated liver injury in rat models. It is well known that the earliest alterations in liver cell structure and function following carbon tetrachloride poisoning involve the endoplasmic reticulum and its drug metabolizing enzymes. Therefore, we investigated whether an aqueous leaf extract of O. aspera could offer protection against carbon tetrachloride-induced changes in the microsomal drug metabolizing enzymes aniline hydroxylase and p aminopyrine N-demethylase. This enzyme activity was compared with phenobarbital induced righting reflex and lipid peroxidation. Treatment of rats with the aqueous leaf extract of O. aspera (before or after the administration of carbon tetrachloride) resulted in a marked decrease in carbon tetrachloride-mediated alterations in aniline hydroxylase and p-aminopyrine N-demethylase activity, phenobarbital-induced loss of righting reflex and malondialdehyde formation due to lipid peroxidation. The Km value of these enzymes in control and Osbeckia treated rats were the same. These results show that the plant extract can markedly decrease the carbon tetrachloride-mediated reduction in aniline hydroxylase and p-aminopyrine N-demethylase activity and inhibit peroxidative damage to the cell membrane. Phenobarbital-induced sleeping time in rats and kinetic enzyme studies suggested that the effects of the plant extract was neither due to an induction of the drug-metabolizing enzymes under investigation, nor due to an alteration in the Km values of these enzymes. PMID- 10813560 TI - Measurement of attitudes towards persons with disabilities. AB - PURPOSE: The purpose of this paper is to assist those engaged in research dealing with attitudes towards persons with disabilities by presenting a catalogue of various attitude measurement methods. METHOD: A review of the methodological and psychological literatures on the measurement of attitudes towards persons with disabilities. RESULTS: The review uncovered 10 direct methods to measure attitudes, in which the respondents are aware that they are participating in an experiment and 14 indirect methods in four categories that are not plagued by attitude-distorting influences because the respondents are not aware that their attitudes are being measured. A discussion of each method with examples is provided, followed by implications for rehabilitation practitioners, rehabilitation education and training, and rehabilitation researchers. CONCLUSIONS: The investigation of attitudes towards persons with disabilities requires innovative experimental methods and psychometrically sound instruments that are reliable, valid, and multidimensional. Without such instruments, it will not be possible to obtain conclusive answers to important research questions concerning the relationship between these attitudes and the acceptance and integration of persons with disabilities into society. PMID- 10813561 TI - A national survey of health-related work limitations among employed persons in the United States. AB - PURPOSE: To estimate the total prevalence of health-related work limitations among working people in the United States (US) as well as their condition specific prevalence. METHODS: A new questionnaire measuring limitations in ability to perform specific work demands was administered to 940 employed people in a national household survey. The prevalence of specific work limitations is reported as are condition-specific risk estimates (odds ratios) based on logistic regression. RESULTS: In the US, 19.3% of working people (CI = 14.0, 24.6) were limited in their abilities to perform physical work demands; 24.1% (CI = 18.9, 29.2) were limited in performing psychosocial work demands; and 13.8% (CI = 8.3, 19.3) were limited in their abilities to function without difficulty within the ambient work environment. With successive increments in the number of conditions, the odds of having a limitation increased significantly. CONCLUSIONS: This study contributes new information concerning the implications of chronic health problems for working people and the significant risks for workers with multiple chronic conditions. PMID- 10813562 TI - The Radboud skills questionnaire: construction and reliability in patients with reflex sympathetic dystrophy of one upper extremity. AB - PURPOSE: To construct a questionnaire to measure skills in which notably both hands are active, which is suitable for patients with a disease of one or both hands and to research its response stability and possible improvements. METHOD: Using the Dutch elaboration of the ICIDH and then experts in a Delphi round, the questionnaire was constructed. Thereafter, test-retest and inter-observer reliability was examined. Fifty-four patients with RSD in one upper extremity and a normal contralateral extremity participated in the study, the first twenty to evaluate the construction of the questionnaire and the others to test reliability. RESULTS: The constructed Radboud skills questionnaire was reliable in terms of response stability (median coefficients of variation 2.2% to 6.6%). Correlation's between categories of items were fair to good. CONCLUSIONS: A useful questionnaire was constructed to map alterations in the level of disability in patients with a disease of one hand or both hands. PMID- 10813563 TI - Access to employment for people with disabilities: findings of a consumer-led project. AB - PURPOSE: A consumer led initiative which aimed to gather information from local employers and disabled people which might inform future action to improve work opportunities for disabled people. METHOD: (1) A survey of 500 companies with more than 20 employees randomly sampled from 4 locations across Suffolk was undertaken. The survey generated both numerical and verbal data. Response rate was low (25%) but achieved a reasonable spread of organizations in terms of size, type of industry and geographical location. (2) Semi-structured interviews were carried out with nine disabled people. Open-ended questions were used to elicit information about the nature of their disability, experiences of education and experiences of seeking and/or maintaining work. RESULTS: 43% of respondents had one or more disabled employees. Evidence of obstacles to employment included a lack of understanding about the capability of disabled people, lack of knowledge about financial and technical assistance and undifferentiated approaches to access and accommodation. Interview data supported the importance of paid employment to self esteem and quality of life and showed high levels of frustration in their search for work. CONCLUSIONS: This study demonstrates the need for better interagency communication and a more effective information distribution strategy for employers, particularly in relation to the availability of systems of support and the capability of disabled employees. PMID- 10813564 TI - Re: Attendant operated wheelchair brakes. PMID- 10813565 TI - Preimplantation genetic diagnosis. AB - Embryo biopsy for preimplantation genetic diagnosis can be performed on the oocyte/zygote, cleavage stage embryo, or blastocyst, but the majority of centres perform cleavage stage biopsy. Single-cell diagnosis is undertaken by the polymerase chain reaction or fluorescent in-situ hybridization. Technical difficulties have arisen with preimplantation genetic diagnosis, such as allele dropout and chromosomal mosaicism. However, it is hoped that these difficulties can be overcome in the future with the advent of new techniques. PMID- 10813566 TI - The application of fluorescence in-situ hybridization to prenatal diagnosis. AB - Fluorescence in-situ hybridization has become essential in prenatal diagnosis for identifying chromosome aberrations as well as in preimplantation genetic diagnosis and the analysis of fetal cells in maternal blood. Comparative genome hybridization, multicolor fluorescence in-situ hybridization and telomere probes provide technical approaches for the characterization of fetal chromosome anomalies not possible by conventional karyotyping. PMID- 10813567 TI - Genetic counselling in prenatally diagnosed non-chromosomal fetal abnormalities. AB - Advances in technology and skills have resulted in the improved detection of fetal ultrasound abnormalities by ultrasound. In addition, the development of new diagnostic methods has resulted in major advances in our ability to detect microscopic and submicroscopic chromosome abnormalities as well as single gene disorders. This often enables us to provide the family with accurate information regarding the aetiology, prognosis, the risk of recurrence and the prenatal diagnosis options available in future pregnancies. Genetic counselling is important because this information should be communicated to the family in simple language, with care and sensitivity, so that the family can make decisions that are fully informed. PMID- 10813568 TI - Amniocentesis and chorionic villus sampling. AB - Invasive prenatal diagnosis continues to be the gold standard for pregnancies at increased risk of chromosomal aneuploidy or other genetic disease. Chorionic villus sampling is the procedure of choice for the first trimester. Early amniocentesis has been shown to carry increased risks of pregnancy loss, amniotic fluid leakage and talipes equinovarus. Mid-trimester amniocentesis continues to be the most common form of invasive prenatal diagnosis, with post-procedural loss rates of between 0.5 and 1%. This present review summarizes information on technique risks, looks at new technology applied to invasive prenatal diagnosis testing, and reports on new diagnoses that could be made either by amniocentesis or chorionic villus sampling. PMID- 10813569 TI - The challenge of prenatal diagnosis in twin pregnancies. AB - Prenatal diagnosis in multiple gestations poses unique problems. The use of screening modalities is limited, diagnostic procedures are more complicated, and the management of discordant results may be required. This article reviews work, both past and present, regarding these issues. PMID- 10813570 TI - Ultrasound of the fetal central nervous system. AB - Current ultrasound equipment allows the antenatal identification of many central nervous system anomalies from early gestation. In selected cases, special techniques (transvaginal sonography, three-dimensional ultrasound, colour Doppler) may enhance the diagnostic potential. Diagnostic accuracy, however, remains heavily dependent upon the expertise of the sonologist. Fetal ultrasound is effective in identifying neural tube defects, although alpha-fetoprotein screening seems to yield a greater sensitivity. The sensitivity in the diagnosis of central nervous system malformations other than neural tube defects remains unclear because of the ascertainment biases of the few large prospective studies that have been carried out so far. Magnetic resonance imaging may play a major role in the evaluation of cases with suboptimal ultrasound visualization, or when specific anomalies are suspected, such as intracranial haemorrhage or migrational disorders. PMID- 10813571 TI - Prenatal diagnosis and management of fetal cardiovascular malformations. AB - Screening for fetal cardiovascular malformations is widely performed. Its accuracy is not yet satisfactory, but better training of ultrasonographers and extension from the four-chamber view to the study of the outflow tract are probably clues to an improvement. The main impact of prenatal diagnosis is still the termination of pregnancy for severe malformations and for those associated with chromosomal or extracardiac anomalies. There is now evidence that prenatal diagnosis improves perinatal morbidity or mortality for some malformations. New information about the molecular genetic basis of congenital heart disease will help in management and counselling. PMID- 10813572 TI - Fetal growth. AB - Recent epidemiological and experimental studies show that abnormal fetal growth can lead to serious complications, including stillbirth, perinatal morbidity and disorders extending well beyond the neonatal period. It is now clear that the intrauterine milieu is as important as genetic endowment in shaping the future health of the conceptus. Maternal characteristics such as weight, height, parity and ethnic group need to be adjusted for, and pathological factors such as smoking excluded, to establish appropriate standards and improve the distinction between what is normal and abnormal. Currently, the aetiology of growth restriction is not well understood and preventative measures are ineffective. Elective delivery remains the principal management option, which emphasizes the need for better screening techniques for the timely detection of intrauterine growth failure. PMID- 10813573 TI - Thromboembolic disease in pregnancy. AB - Recent publications have produced some new estimates of the incidence of pregnancy-related venous thromboembolic disease, and have found increasing evidence of an association between inherited thrombophilias and pregnancy complications and fetal loss. The balance of benefit and risk of thromboprophylaxis remains to be evaluated, and studies are needed to provide a sound basis for clinical practice. PMID- 10813574 TI - Intrapartum fetal surveillance. AB - Intrapartum hypoxia was thought to contribute to the incidence of cerebral palsy, seizures and mental retardation. Electronic fetal monitoring was expected to prevent or reduce this incidence. Electronic fetal monitoring has a high false positive rate and fetal blood sampling, which is an invasive procedure, only allows an intermittent assessment. Efforts are being made to improve fetal heart rate analysis and clinical management. Fetal pulse oximetry, fetal electrocardiogram waveform analysis and the intermittent measurement of lactate levels by fetal blood sampling may become established as an adjunct to electronic fetal monitoring. PMID- 10813575 TI - Amnioinfusion for meconium-stained liquor. AB - Amnioinfusion reduces the risk of meconium aspiration by the infants of women with thick meconium staining of the amniotic fluid. The benefits are clear in facilities with high baseline rates of meconium aspiration, and are therefore likely to outweigh the risk of uncommon but serious maternal side-effects. Larger randomized trials are needed to determine more precisely the relative risks and benefits in facilities with low baseline rates of meconium aspiration. The addition of antibiotics to the infusate has not been shown to reduce the risk of sepsis related to meconium. PMID- 10813576 TI - Bibliography. Current world literature. Maternal-fetal medicine and prenatal diagnosis. PMID- 10813577 TI - p53--the link between inflammation and cancer? PMID- 10813578 TI - Fetal origin of leukemia and autologous cord blood transfusions. PMID- 10813579 TI - Reduced uterine and ovarian size in adolescent girls born small for gestational age. AB - Reduced fetal growth is known to be associated with a reduced ovarian fraction of primordial follicles, with ovarian hyperandrogenism and anovulation in late adolescence. In this study, we examined whether adolescent girls born small for gestational age also present an abnormality in uterine or ovarian size. Standardized ultrasound measurements of the internal genitalia were performed in 36 healthy post-menarcheal girls (mean age 14 y) born with a size that was either appropriate for gestational age (AGA) or small (SGA), birth weight averaging 0.1 and -3.0 SD, respectively; clinical and endocrine characteristics were documented concomitantly. Compared with AGA girls, the SGA girls had a smaller uterus (mean difference of 20%; p < 0.006) and a reduced ovarian volume (mean difference of 38%; p < 0.0002). In conclusion, the gynecological correlates of prenatal growth restriction are herewith extended to include a reduced size of the uterus and the ovaries. PMID- 10813580 TI - Body composition during the first 2 years of life: an updated reference. AB - Normative body composition during the first 2 y of life was derived from a prospective study of 76 children. We present 1) fat free mass (FFM) and its components, and fat mass (FM), 2) incremental growth rates partitioned into chemical components, and 3) age-specific and gender-specific constants for converting chemical and physical components into FFM for children during the first 2 y of life. A multicomponent model based on measurements of total body water (TBW), total body potassium (TBK) and bone mineral content (BMC) was used to estimate FFM and FM at 0.5, 3, 6, 9, 12, 18, and 24 mo of age. TBW was determined by deuterium dilution, TBK by whole body counting, and BMC by dual energy x-ray absorptiometry. FFM was higher in boys than girls between 0.5-18 mo of age (p < or = 0.05). Percent FM increased on average from 13 to 31% between 0.5 and 3-6 mo, and then gradually declined. Percent FM was significantly higher in girls than in boys at 6 and 9 mo of age (p < or = 0.02). The components of FFM on a percentage basis changed with age (p = 0.001), but not gender. The protein content of FFM increased gradually with age, while TBW declined (p = 0.001). As a percentage of FFM, osseous mineral increased from 2.0 to 3.4% in boys and from 2.1 to 3.3% in girls between 0.5 and 24 mo (p = 0.001). Density and potassium content of FFM increased gradually with age (p = 0.001). These normative body composition data provide an updated reference upon which to assess normal growth and nutritional status of pediatric populations representative of mixed feeding groups during the first 2 y of life. PMID- 10813581 TI - Cranial irradiation of female rats causes dose-dependent and age-dependent activation or inhibition of pubertal development. AB - Cranial irradiation in prepubertal children with leukemia or brain tumors can lead to precocious or in high doses to late puberty. To unravel the underlying mechanisms, we developed a rat model with selective cranial Co60-irradiation technique. Infantile (12-16 d old) or juvenile (21-23 d old) female Sprague Dawley rats received a single dose of 4, 5, 6, 9 or 2 x 9 Gy (at days 21 and 23). Each group consisted of 7-20 animals. High radiation doses (9 Gy and more) caused retardation of sexual development, whereas low radiation doses (5 or 6 Gy) led to accelerated onset of puberty in 20% of infantile irradiated rats animals as determined by vaginal opening. Interestingly, at peripubertal age (postnatal day 32-34), 5 or 6 Gy infantile irradiated rats had significantly higher serum LH levels stimulated by GnRH and estradiol levels (p < 0.05). 2 x 9 Gy irradiated rats had at the age of 3 mo a marked growth retardation and significantly lower GH levels than the controls (p < 0.05) whereas prolactin, FSH, TSH, T4, and corticosterone levels were comparable with controls. These studies demonstrate that the GnRH-pulse generator is very radiosensitive as precocious activation occurred after low dose irradiation (5 or 6 Gy) of infantile rats without any other endocrine disorder. High radiation doses (9 or 2 x 9 Gy) induced retardation of sexual maturation and later on growth hormone deficiency. Moreover this model of cranial irradiation seems to be suitable to study the molecular mechanisms of radiation induced pubertal changes. PMID- 10813582 TI - Chemoattractant factors in breast milk from allergic and nonallergic mothers. AB - The allergy-preventing effect of breast-feeding remains controversial, possibly because of individual variations in the composition of the breast milk. Recently, we showed that allergic mothers had higher concentrations of IL-4 and lower concentrations of ovalbumin-specific IgA in their breast milk than nonallergic mothers. The aim of this study was to investigate the concentrations of chemokines and cytokines that are chemotactic to cells involved in allergic reactions in breast milk from allergic and nonallergic mothers. Cytokine and chemokine concentrations were determined with ELISA in colostrum and mature milk samples from 23 mothers with and 25 mothers without atopic symptoms. IL-8 was detected in all milk samples. RANTES (regulated on activation, normal T cell expressed and secreted), eotaxin, and IL-16 were detected in 50%, 76%, and 48%, respectively, in colostrum and less commonly in mature milk. Macrophage inflammatory protein-1alpha, however, could not be detected in any of the samples. The concentrations of IL-8 and RANTES were higher in breast milk from allergic, compared with nonallergic, mothers. In conclusion, the presence of chemoattractant factors in breast milk may be responsible for the traffic of leukocytes from the maternal circulation to the breast milk. The higher concentrations of RANTES and IL-8 in allergic mothers may partly explain the controversy regarding the protective effect of breast-feeding against the development of allergy by stronger chemotaxis and activation of cells involved in allergic diseases, and possibly by elevated IgE production. PMID- 10813583 TI - Parental smoking and neonatal serum levels of polychlorinated biphenyls and hexachlorobenzene. AB - Polychlorinated biphenyls (PCB) and hexachlorobenzene (HCB) are ubiquitous compounds that have tumor-promoting properties if applied together with tobacco specific carcinogens. It was the purpose of the present study to investigate whether parental smoking by itself will increase the prenatal uptake of such organochloric compounds. With the informed consent of the parents, blood samples were taken from 80 full-term neonates before the first oral feeding. Six PCB congeners (PCB 28, 52, 101, 138, 153, and 180) and HCB were analyzed with capillary gas chromatography. Information about parental smoking behavior, the geographic origin of the parents, and their actual and previous working places was recorded. We composed three study groups for statistical analyses: active smoking mothers (n = 12), passive smoking mothers (n = 33), and nonsmoking families (n = 35). Neonates born to active smoking mothers had the highest PCB and HCB concentrations compared with children of passive or nonsmoking mothers. These differences were statistically significant (p < 0.01) in the cases of PCB 138, total PCB, and HCB. Newborns of passive smoking mothers had higher PCB and HCB concentrations than children of nonsmoking families but lower values than those of active smoking mothers. These differences were statistically significant for all compounds with the exception of PCB 180. It is concluded that active and passive maternal smoking increases the neonatal burden with PCB and HCB. PMID- 10813584 TI - Cyclooxygenase-2 in human perinatal lung. AB - Cyclooxygenases-1 and -2 are the key enzymes in the conversion of arachidonic acid to prostanoids. Cyclooxygenase-2 (COX-2) takes part both in inflammation and in control of cell growth. COX-2 immunohistochemistry was performed on lung tissues from autopsies, with four groups included: fetuses (n = 4, GA = 16.0 to 32.0 wk), preterm infants (n = 10, GA = 23.0 to 29.9 wk), term infants (n = 6, GA = 38.7 to 42.0 wk), and infants with bronchopulmonary dysplasia (BPD) (n = 4, GA = 28.9 to 30.7 wk). COX-2 staining occurred exclusively in the epithelial cells resembling type II pneumocytes in the alveolae, and in ciliated epithelial cells in the bronchi. In fetuses, moderate intensity alveolar staining was seen in 90 100% cells lining the alveolar epithelium. In preterm infants, high intensity alveolar staining was seen in a scattered pattern. In term infants, the alveolar staining was also scattered, but with a lower proportion of positive cells. In BPD no staining appeared in alveolar epithelial cells. The most intense bronchial staining was found in fetuses and the least intense in term infants; staining was also seen in BPD. COX-2 is present in human perinatal lung from the gestational age of 16 wk, in a changing pattern. We suggest that COX-2 may, in addition to participating in inflammation, also play a developmental role in the perinatal lung. PMID- 10813585 TI - Expression of vascular endothelial growth factor and Flk-1 in developing and glucocorticoid-treated mouse lung. AB - Although the endothelial cell is the most abundant cell type in the differentiated lung, little is known about regulation of lung developmental vasculogenesis. Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen and angiogenic factor that has putative roles in vascular development. Mitogenic actions of VEGF are mediated by the tyrosine kinase receptor KDR/murine homologue fetal liver kinase Flk-1. HLF (hypoxia-inducible factor-like factor) is a transcription factor that increases VEGF gene transcription. Dexamethasone augments lung maturation in fetal and postnatal animals. However, in vitro studies suggest that dexamethasone blocks induction of VEGF. The objectives for the current study were to measure VEGF mRNA and Flk-1 mRNA in developing mouse lung and to measure the effects of dexamethasone treatment in vivo on VEGF and Flk-1 in newborn mouse lung. Our results show that VEGF and Flk-1 messages increase in parallel during normal lung development (d 13 embryonic to adult) and that the distal epithelium expresses VEGF mRNA at all ages examined. Dexamethasone (0.1-5.0 mg x kg(-1) x d(-1)) treatment of 6-d-old mice resulted in significantly increased VEGF, HLF, and Flk-1 mRNA. Dexamethasone did not affect cell-specific expression of VEGF, VEGF protein, or proportions of VEGF mRNA splice variants. These data suggest that the developing alveolar epithelium has an important role in regulating alveolar capillary development. In addition, unlike effects on cultured cells, dexamethasone, even in relatively high doses, did not adversely affect VEGF expression in vivo. The relatively high levels of VEGF and Flk-1 mRNA in adult lung imply a role for pulmonary VEGF in endothelial cell maintenance or capillary permeability. PMID- 10813586 TI - Angiotensin II and basic fibroblast growth factor mitogenic pathways in human fetal mesangial cells. AB - Angiotensin II (Ang II) and basic fibroblast growth factor (bFGF/FGF-2) play relevant roles in renal development. Since the signaling pathways modulating the mitogenic effects of Ang II and bFGF in human fetal mesangial cells (HFMc) are not clearly defined, we carried out experiments to determine whether they would exert their mitogenic effects by modulating the activity of the mitogen-activated protein kinases (MAPK) [extracellular signal-regulated kinase-2 (ERK-2)] and cAMP signaling pathways. In confluent HFMc, bFGF (20 ng/mL) induced a significant 4 fold increase in ERK-2 activity and [3H]-thymidine incorporation (6-fold). In contrast, under similar tissue culture conditions, Ang II (10(-6) M) induced a more modest increase in ERK-2 activity (2-fold) and [3H]-thymidine incorporation (35 +/- 4%). The mitogen-activated protein kinase kinase-1 (MEK-1) inhibitor PD098059 (25 microM) almost completely abolished the bFGF-induced proliferation in HFMc but did not significantly affect Ang II proliferative effects. In the presence of the cAMP elevating agent isoproterenol, Ang II and bFGF induced opposite changes in cAMP accumulation and cell growth. Isoproterenol inhibited the basal and bFGF-induced proliferation of HFMc through a MEK-1/2-independent pathway that included the accumulation of cAMP. In contrast, isoproterenol increased Ang II mitogenic effects in correlation with a reduction in cAMP accumulation. We conclude that Ang II and bFGF modulate the proliferation of HFMc through the stimulation of different MEK-1/2-dependent and independent signaling pathways. Activation of MEK-1/2 is required but not sufficient for mitogenesis in HFMc. The accumulation of cAMP in HFMc counteracts the mitogenic effects of bFGF by a MEK-1/2-independent pathway. PMID- 10813587 TI - Antenatal glucocorticoids alter postnatal preterm lamb renal and cardiovascular responses to intravascular volume expansion. AB - We assessed renal and cardiovascular function in preterm newborn lambs after antenatal glucocorticoid exposure. Pregnant ewes were randomly assigned to receive betamethasone or saline via either direct fetal or maternal injection at 122 d gestation. Lambs were delivered 15 h later, and cardiovascular and renal function was assessed. Two hours after delivery, baseline urine flow, urinary sodium excretion, and urinary osmolar clearance were similar in all groups. Volume expansion (saline, 2.5% of body weight, for 10 min) increased values for urine flow (0.23 +/- 0.04 to 0.58 +/- 0.09 mL x min(-1) x kg(-1)), urinary sodium excretion (29.7 +/- 5.8 to 76.2 +/- 12.3 microEq x min(-1) x kg(-1)), and osmolar clearance (12.2 +/- 1.2 to 24.3 +/- 1.6 mL/100 mL GFR) in the fetal group. Increases in urine values were also observed in the maternal group, but control values did not change significantly. Mean arterial pressure was increased in both betamethasone-treated groups relative to controls. Short-term antenatal betamethasone exposure 1) augments preterm newborn kidney adaptive responses to acute volume expansion, and 2) increases postnatal blood pressure in preterm newborn lambs. PMID- 10813588 TI - Augmentation of alimentary insulin secretion despite similar gastric inhibitory peptide (GIP) responses in juvenile obesity. AB - Insulin secretion rates are greater after oral glucose than after parenteral administration of an equivalent glucose load. This augmented beta-cell secretory response to an oral glucose load results from the release of mainly two gut hormones: gastric inhibitory polypeptide (GIP) and glucagon-like peptide-1, which potentiate glucose-induced insulin secretion. Because of their insulinotropic action, their abnormal secretion may be involved in the pathogenesis of the hyperinsulinemia of childhood obesity. In this study, we used the hyperglycemic clamp with a small oral glucose load to assess the effect of childhood obesity on GIP response in seven prepubertal lean and 11 prepubertal obese children and in 14 lean adolescents and 10 obese adolescents. Plasma glucose was acutely raised to 11 mM by infusing i.v. glucose and kept at this concentration for 180 min. Each subject ingested oral glucose (30 g) at 120 min, and the glucose infusion was adjusted to maintain the plasma glucose plateau. Basal insulin and C-peptide concentrations and insulin secretion rates (calculated by the deconvolution method) were significantly greater in obese children compared with lean children (p < 0.001). Similarly, during the first 120 min of the clamp, insulin secretion rates were higher in obese than lean children. After oral glucose, plasma insulin, C-peptide, and insulin secretion rates further increased in all four groups. This incretin effect was 2-fold greater in obese versus lean adolescents (p < 0.001). Circulating plasma GIP concentrations were similar at baseline in all four groups and remained unchanged during the first 120 min of the clamp. After oral glucose, plasma GIP concentrations rose sharply in all groups (p < 0.002). Of note, the rise in GIP was similar in both lean and obese children. In conclusion, under conditions of stable hyperglycemia, the ingestion of a small amount of glucose elicited equivalent GIP responses in both lean and obese children. However, despite similar GIP responses, insulin secretion was markedly augmented in obese adolescents. Thus, in juvenile obesity, excessive alimentary beta-cell stimulation may be independent of the increased release of GIP. PMID- 10813589 TI - A study of chromosomal aberrations and chromosomal fragility after recombinant growth hormone treatment. AB - Increased chromosomal rearrangements and chromosomal fragility have been previously observed in lymphocytes of children treated with human GH, implying that treatment could predispose to malignancy. Twenty-four children with classic GH deficiency, neurosecretory GH dysfunction, and Turner syndrome were treated with recombinant human GH (0.3 mg x kg(-1) x wk(-1)). Metaphase cells were assessed for spontaneous chromosomal and chromatid aberrations at baseline and 6 mo into treatment. There were no significant differences in aberrations between baseline and the 6-mo samples. However, the mean frequency of chromatid-type aberrations on a per cell basis was significantly higher than at baseline, 0.0088 versus 0.0064 aberrations per cell (p < 0.024). Two patients contributed inordinately to this increase. A third sample from these two patients was almost identical to their baseline samples. Cells were also irradiated in vitro (3 Gy) to assess chromosomal fragility. After irradiation, no patient showed a significant difference for any aberration type, although there was a significantly lower frequency of ring chromosomes on a per cell basis in the 6-mo samples (p < 0.001). We find no evidence that GH therapy influences spontaneous chromosomal aberrations or chromosomal fragility. PMID- 10813591 TI - Nicotine delays arousal during hypoxemia in lambs. AB - A decreased ability to arouse from sleep in response to arterial hypoxemia may lead to severe asphyxia and has been proposed as a mechanism of sudden infant death syndrome. Based on previous observations that nicotine exposure, a major environmental risk factor for sudden infant death syndrome, may impair hypoxic defense in neonates, we hypothesized that a short-term infusion of nicotine could impair hypoxic arousal through interference with oxygen-sensing mechanisms. Seven chronically instrumented unanesthetized lambs were studied at the age of 4.6 +/- 1.3 d during normoxia and acute hypoxia (0.1 fraction of inspired oxygen) for 5 min. Ventilation, transcutaneous Hb oxygen saturation, blood pressure, heart rate, and time to arousal were compared during a control saline infusion and during a 0.5 microg x kg(-1) x min(-1) nicotine infusion. Activity states, i.e. wakefulness and quiet sleep as well as arousal, were defined by EEG, nuchal electromyogram, and electrooculogram. Each lamb acted as its own control. Arousal from quiet sleep occurred significantly later during nicotine infusion compared with control (177 +/- 93 versus 57 +/- 41 s, p < 0.01) and at a lower transcutaneous Hb oxygen saturation (60 +/- 12 versus 79 +/- 12%, p < 0.01) (paired t test). The ventilatory response to hypoxia in wakefulness was similar during both conditions but was significantly attenuated in quiet sleep during nicotine infusion (p < 0.001, 2-way ANOVA repeated-measures design). Blood pressure and heart rate responses were similar during both conditions. These results suggest that a brief nicotine exposure blunts oxygen sensitivity in young lambs, a finding of potential relevance for sudden infant death syndrome. PMID- 10813590 TI - Elevated protein carbonyls and lipid peroxidation products correlating with myeloperoxidase in tracheal aspirates from premature infants. AB - The purpose of this study is to determine whether the oxidative injury markers, protein carbonyls and malondialdehyde (MDA), are elevated in tracheal aspirates from very low birth weight (< 1500 g) infants; to determine whether levels correlate with myeloperoxidase as a marker of neutrophil inflammation; and to assess whether high levels are associated with poor respiratory outcome. Tracheal aspirates (144 samples) were collected from 86 infants < 1500 g at times of routine suctioning. Aspirates (82 samples) from 54 infants > or = 1500 g who required intubation for a variety of diagnoses were analyzed for comparison. Analyses were performed for protein carbonyls by ELISA, total malondialdehyde by HPLC, and myeloperoxidase activity. Respiratory outcome was assessed as oxygen requirement at 28-d or 36-wk postmenstrual age, and as the number of days of oxygen requirement. Protein carbonyls were significantly higher in infants < 1500 g than larger infants, and were highest close to birth. MDA concentrations were also higher in the earlier samples. There was a strong positive correlation between protein carbonyls and myeloperoxidase, suggesting a link between protein oxidation and neutrophil activation. A similar but weaker correlation was seen for MDA. Carbonyls in samples taken after steroid administration were less than for controls with a similar age distribution. We did not see significant associations between oxidant marker levels and development of chronic lung disease. Our findings of higher amounts of protein and lipid oxidation products in tracheal aspirates with high myeloperoxidase activity, taken together with other studies showing a link between neutrophil accumulation and chronic lung disease, suggest a possible contribution by neutrophil-derived reactive oxygen species to the injury. PMID- 10813592 TI - Cardiopulmonary exercise performance in prematurely born children. AB - Prematurely born children have reduced peak VO2 compared with their peers, inferentially attributed to ventilatory limitation. The primary purpose of this study was to compare exercise ventilation and cardiac output in a sample of childhood survivors of lung disease of prematurity with those of a control group to elucidate reasons for lower peak VO2. A secondary aim was to describe and compare the ventilatory response to incremental exercise. Thirty-two children, aged 8-9 y, were recalled for lung function and progressive exercise tests. Fifteen of them also performed submaximal exercise with measurement of cardiac output (indirect [CO2] Fick) and physiologic dead space. Results were compared with those of term-born, age- and sex-matched, control children. Pulmonary function tests showed mild airflow limitation. Peak VO2 was lower in prematurely born children compared with control children, and was correlated with lean body mass. Their heart rate-VO2 relationship and stroke volume were similar to that of term-born control children. Children with a history of bronchopulmonary dysplasia and hyaline membrane disease as infants exhibited greater exercise hyperpnea than did healthy control children, because of higher breathing frequency, and maintained lower end-tidal PCO2 during submaximal exercise. Physiologic dead space normalized for body weight was similar in preterm and term-born children. Lower peak VO2 in this population is not caused by cardiopulmonary factors, but is best predicted by lean body mass. Ventilation did not limit exercise performance, although it appears that breathing during exercise is regulated differently in prematurely born children than in term-born children. PMID- 10813593 TI - Spectral analysis of heart rate variability in premature infants with feeding bradycardia. AB - An elevated level of baseline parasympathetic activity was noted in a group of premature infants suffering from bradycardia during feeding. At approximately 34 wk post-conceptional age, the heart rates of 12 infants with feeding bradycardia (birth weight = 1539 +/- 279 g; gestational age = 31.0 +/- 1.6 wk) and 10 controls (birth weight = 1710 +/- 304 g; gestational age = 32.0 +/- 1.4 wk) were recorded 1 h before and 1 h after feeding. EKG data were digitized and 3.2-min segments of data were analyzed to determine the spectral power at very low (VLF = 0.003-0.03 Hz), low (LF = 0.03-0.39 Hz), and high (HF = 0.40-1.00 Hz) frequencies. In preterm infants with feeding bradycardia, an elevation in baseline parasympathetic activity was evident before feeding, as indicated by significantly higher HF power and a lower LF/HF ratio. This elevation in baseline parasympathetic activity may contribute to the observed bradycardia during feeding. PMID- 10813594 TI - Effects of GABA receptor blockade on the ventilatory response to hypoxia in hypothermic newborn piglets. AB - Hypothermic newborn piglets have a depressed ventilatory response to hypoxia, and this may be due to an increase in CNS gamma-aminobutyric acid (GABA) levels. To evaluate the effects of GABA(A) receptor blockade on the ventilatory response to hypoxia in hypothermic piglets, 31 anesthetized paralyzed mechanically ventilated newborn piglets (2-7 d) were studied at a brain temperature of 38.5 +/- 0.5 degrees C [normothermia (NT), n = 15] or 34 +/- 0.5 degrees C [hypothermia (HT), n = 16]. The central respiratory output was evaluated by measuring burst frequency and moving time average area of phrenic nerve activity. Measurements of minute phrenic output (MPO), arterial blood pressure, heart rate, oxygen consumption, and arterial blood gases were obtained at room air and during 20 min of isocapnic hypoxia [fraction of expired oxygen (FiO2) = 0.10]. After 10 min of hypoxia, a bolus injection of 20 microL of bicuculline methiodide (BM; 10 microg) or Ringer's solution was administered into the cisterna magna over a 1-min period, and the piglets remained in hypoxia for an additional 10 min. There was an initial increase of 50 +/- 6% in MPO during the first minute of hypoxia followed by a decrease to values 24 +/- 8% above baseline at 10 min in the NT group. In contrast, in the HT group, the initial increase in MPO with hypoxia was eliminated, and, at 10 min, there was a decrease to a mean value 35 +/- 4% below baseline level (NT versus HT, p < 0.03). After administration of BM, a significant increase in MPO with hypoxia was observed in both groups compared with their placebo groups (p < 0.002 in NT-BM group, p < 0.0001 in HT-BM group). However, the magnitude of the increase in MPO during hypoxia was significantly greater in the HT group after administration of BM (NT versus HT, p < 0.0001). Changes in oxygen consumption, arterial blood pressure, heart rate, pH, partial pressure of oxygen (PaO2), and base excess with hypoxia were not different between NT and HT groups before and after the administration of BM. The cardiorespiratory response to hypoxia was not modified after administration of Ringer's solution to NT and HT placebo groups. These data suggest that the depression in hypoxic ventilatory response produced by HT is in part modulated by an increased CNS GABA concentration. PMID- 10813595 TI - Increased cardiomyocyte intracellular calcium during endotoxin-induced cardiac dysfunction in guinea pigs. AB - Septic shock is a complex pathophysiologic state characterized by circulatory insufficiency, multiple system organ dysfunction, and frequent mortality. Although profound cardiac dysfunction occurs during sepsis, the pathogenesis of this dysfunction remains poorly understood. To determine whether abnormalities in intramyocyte calcium accumulation might contribute to the development of cardiac dysfunction, we measured myocyte intracellular calcium during peak cardiac dysfunction after an endotoxin challenge. Intraperitoneal administration of Escherichia coli lipopolysaccharide 4 mg/kg to adult guinea pigs resulted in significantly impaired cardiac performance (Langendorff preparation) 18 h after challenge compared with control. This included diminished left ventricular pressure development (56 +/- 7 versus 95 +/- 4 mm Hg, p < 0.05), maximal rate of left ventricular pressure rise (998 +/- 171 versus 1784 +/- 94 mm Hg/s, p < 0.05) and left ventricular pressure fall (1014 +/- 189 versus 1621 +/- 138 mm Hg/s, p < 0.05). Assay of intracellular calcium in fura-2AM-loaded cardiac myocytes demonstrated increased intracellular calcium concentration in myocytes obtained from lipopolysaccharide-challenged animals compared with controls (234 +/- 18 versus 151 +/- 6 nM, p < 0.05). Inhibition of calcium-release channel (ryanodine receptor) opening by administration of dantrolene prevented the increase in intracytoplasmic calcium (159 +/- 8 versus 234 +/- 18 nM, p < 0.05) and partially ameliorated systolic and diastolic ventricular dysfunction. These data indicate that abnormalities of intracellular calcium contribute to the development of endotoxin-induced myocardial dysfunction. PMID- 10813596 TI - Ontogenesis of CYP2C-dependent arachidonic acid metabolism in the human liver: relationship with sudden infant death syndrome. AB - A modification of the human monooxygenase system have been previously associated with the sudden infant death syndrome (SIDS): the hepatic CYP2C content was markedly enhanced and resulted from an activation of CYP2C gene transcription. To determine the possible consequence of the up-regulation of CYP2C in SIDS, we examined the metabolism of arachidonic acid (AA) an endogenous substrate of CYP2C involved in the physiologic regulation of vascular tone. The overall AA metabolism was extremely low during the fetal period and rose after birth to generate 14,15 epoxyeicosatrienoic acid (EET), 11,12 EET and the sum of 5,6 dihydroxyeicosatrienoic acid (diHETE)+omega/omega-1 hydroxy AA. In SIDS, the accumulation of CYP2C proteins was associated with a significant increase in the formation of 14,15 and 11,12 diHETE, which were shown to be supported by individually expressed CYP2C8 and 2C9 and HETE1 (presumably 15 HETE). This increase was markedly inhibited by addition of sulfaphenazole, a selective inhibitor of CYP2C9. So, we propose that the higher CYP2C content in SIDS stimulates the production of EETs and diHETEs and might have severe pathologic consequences in children. PMID- 10813597 TI - Low doses of dexamethasone suppress pituitary-adrenal function but augment the glycemic response to acute hypoxemia in fetal sheep during late gestation. AB - Despite the widespread use of antenatal glucocorticoid therapy in obstetric practice, little is known about the effects of synthetic glucocorticoids on the fetal capacity to respond to episodes of acute hypoxemia, such as may occur during labor and delivery. This study investigated the effects of prolonged fetal exposure to low concentrations of dexamethasone on the fetal ACTH, cortisol, and glycemic responses to an episode of acute hypoxemia during the period of dexamethasone treatment in sheep. At 118 d of gestation (term is approximately 145 d), 11 fetal sheep had catheters implanted under halothane anesthesia. From 124 d, five fetuses were infused i.v. continuously with dexamethasone (1.80 +/- 0.15 microg x kg(-1) x h(-1) in 0.9% saline at 0.5 mL/h) for 48 h, and the other six fetuses received saline solution i.v. at the same rate. At 45 h of infusion, acute hypoxemia was induced in all fetuses for 1 h by reducing the maternal inspired fraction of oxygen. During glucocorticoid treatment, fetal plasma dexamethasone concentrations increased to 3.9 +/- 0.2 nM by 24 h and remained elevated for the rest of the infusion period. During hypoxemia, a similar fall in fetal arterial PO2 occurred in both saline-infused and dexamethasone-treated fetuses. In control fetuses, significant increases in plasma ACTH and cortisol concentrations and in blood glucose concentrations occurred during hypoxemia. Dexamethasone treatment prevented the increases in fetal plasma ACTH and cortisol, and augmented the blood glucose response, induced by hypoxemia. These data indicate that prolonged fetal exposure to low concentrations of dexamethasone suppresses pituitary-adrenal function, but augments the glycemic response, to acute hypoxemia in fetal sheep during late gestation. PMID- 10813598 TI - Dietary supplementation of arachidonic acid is associated with higher whole body weight and bone mineral density in growing pigs. AB - Essential fatty acids are fundamental to normal growth and development, but North American formulas do not contain arachidonic (AA) and docosahexaenoic acid (DHA). The main objective of the present study was to determine whether addition of AA and DHA to formula elevates growth and bone mineralization in piglets. A secondary objective was to establish whether liver fatty acid composition is related to that of bone. Twelve 10-d-old male piglets were randomized to receive either a standard formula with an n-6:n-3 fatty acid ratio of 4.9:1.0 or the same formula made with an equal amount of fat but containing AA (0.5% wt/wt total fat) and DHA (0.1% wt/wt total fat) for 14 d. Piglets in the supplemented group had significantly (p < 0.05) higher weight and greater bone mineral density of the whole body, lumbar spine, and femur. No differences were observed in whole body length, calcium absorption, or biochemical markers of bone metabolism. Feeding AA resulted in lower linoleic acid (p < 0.05) and higher (p < 0.05) AA in liver total lipid (% wt/wt) and bone FFA (% wt/wt) but no change to DHA. Liver AA (% wt/wt total lipid) was positively related (p < 0.05) to growth, free AA (% wt/wt) in bone, bone mineral content, bone mineral density, and urinary prostaglandin E2 but negatively related (p < 0.05) to free linoleic acid in bone. Inverse relationships were observed when liver linoleic acid was substituted for liver AA as the independent variable. These data indicate that feeding AA is associated with elevated weight and higher whole body and regional bone mineral density. PMID- 10813599 TI - Pathogenicity of avian malaria in experimentally-infected Hawaii Amakihi. AB - The introduction of avian malaria (Plasmodium relictum) and mosquitoes (Culex quinquefasciatus) to the Hawaiian Islands (USA) is believed to have played a major role in the decline and extinction of native Hawaiian honeycreepers (Drepanidinae). This introduced disease is thought to be one of the primary factors limiting recovery of honeycreepers at elevations below 1,200 m where native forest habitats are still relatively intact. One of the few remaining species of honeycreepers with a wide elevational distribution is the Hawaii Amakihi (Hernignathus virens). We measured morbidity and mortality in experimentally-infected Hawaii Amakihi that were captured in a high elevation, xeric habitat that is above the current range of the mosquito vector. Mortality among amakihi exposed to a single infective mosquito bite was 65% (13/20). All infected birds had significant declines in food consumption and a corresponding loss in body weight over the 60 day course of the experiment. Gross and microscopic lesions in birds that succumbed to malaria included enlargement and discoloration of the spleen and liver and parasitemias as high as 50% of circulating erythrocytes. Mortality in experimentally-infected amakihi was similar to that observed in Apapane (Himnatione sanguinea) and lower than that observed in Iiwi (Vestiaria coccinea) infected under similar conditions with the same parasite isolate. We conclude that the current elevational and geographic distribution of Hawaiian honeycreepers is determined by relative susceptibility to avian malaria. PMID- 10813600 TI - Fatal toxoplasmosis in free-ranging endangered 'Alala from Hawaii. AB - The 'Alala (Corvus hawaiiensis) is the most endangered corvid in the world, and intensive efforts are being made to reintroduce it to its former native range in Hawaii. We diagnosed Toxoplasma gondii infection in five free-ranging 'Alala. One 'Alala, recaptured from the wild because it was underweight and depressed, was treated with diclazuril (10 mg/kg) orally for 10 days. Antibodies were measured before and after treatment by the modified agglutination test (MAT) using whole T. gondii tachyzoites fixed in formalin and mercaptoethanol. The MAT titer decreased four-fold from an initial titer of 1:1,600 with remarkable improvement in physical condition. Lesions of toxoplasmosis also were seen in two partially scavenged carcasses and in a third fresh intact carcass. Toxoplasma gondii was confirmed immunohistochemically by using anti-T. gondii specific serum. The organism was also cultured by bioassay in mice from tissues of one of these birds and the brain of a fifth 'Alala that did not exhibit lesions. The life cycle of the parasite was experimentally completed in cats. This is the first record of toxoplasmosis in 'Alala, and the parasite appears to pose a significant threat and management challenge to reintroduction programs for 'Alala in Hawaii. PMID- 10813601 TI - Agglutinating antibodies to Toxoplasma gondii in sera from captive eastern barred bandicoots in Australia. AB - Toxoplasmosis is considered a severe health risk for many marsupial species. The mainland Australian population of bandicoot is endangered. Therefore, a preliminary serosurvey was conducted to evaluate exposure to Toxoplasma gondii in 57 captive eastern barred bandicoot and to estimate the possible impact of Toxoplasma on recovering populations. Five (9%) bandicoot were classified as seropositive using a modified agglutination test. Nineteen additional bandicoot (33%) were classified as serosuspect using a direct agglutination test. No bandicoot showed signs of clinical disease. Seropositive titers were IgG associated, suggesting that infections were chronic and latent. Serostatus was not associated with either sex or being wild-caught, although each seropositive bandicoot was wild-caught. Seropositive animals ranged from 1.25- to 2.5-yr-old. Computer simulations using Vortex 5.1, based on the proportion of seropositive and seronegative bandicoot in this study, indicate that mortalities from T. gondii should have little impact upon captive populations. However, the potential impact of toxoplasmosis on recovery efforts for wild, mainland bandicoot populations is not clear. PMID- 10813602 TI - Serologic survey for Toxoplasma gondii in selected wildlife species from Alaska. AB - Blood was collected from selected wildlife species in specific areas of Alaska (USA) during 1976-96. A modified agglutination test was used to test sera for evidence of exposure to Toxoplasma gondii. Serum antibody prevalence was 43% (62 positive of 143 tested) for black bears (Ursus americanus), 9% (11/125) for wolves (Canis lupus), 7% (22/319) for Dall sheep (Ovis dalli), 6% (14/241) for caribou (Rangifer tarandus), 1% (3/240) for moose (Alces alces), and 1% (2/241) for bison (Bison bison). A predictive model was developed to determine the effect of sex, age, location, and year of collection on antibody prevalence for each species. Prevalence was higher in older black bears, caribou, and wolves. For black bears, prevalence was highest in the southeast region of the state. For caribou, prevalence was lowest on the Alaska Peninsula. PMID- 10813603 TI - Characterization of poxviruses from forest birds in Hawaii. AB - Two strains of avian pox viruses were isolated from cutaneous lesions in Hawaiian crows (Corvus hawaiiensis) examined in 1994 and a third from a biopsy obtained in 1992 from an infected bird of the Apapane species (Himatione sanguinea) by inoculation of the chorioallantoic membranes (CAM) of developing chicken embryos. The resulting proliferative CAM lesions contained eosinophilic cytoplasmic inclusion bodies characteristic of pox virus infection. The pathogenicity of these three viruses in domestic chickens was mild as evidenced by the development of relatively minor lesions of short duration at the sites of inoculation. Their virulence in this host was similar to that of a fowlpox virus (FPV) vaccine strain and contrasted greatly with the ability of two field strains of FPV to produce extensive proliferative lesions. One of the Hawaiian crow pox virus isolates as well as the one originating from the Apapane species could be propagated in two secondary avian cell lines, QT-35 and LMH. A comparison of the restriction fragment length polymorphisms (RFLP) of the genomes of the two cell line-adapted viruses, generated by EcoRI digestion, revealed a limited degree of similarity. Moreover, neither profile was comparable to those of the two field isolates of FPV, which were almost indistinguishable from each other. Thus, based on the genetic distinctness of the two Hawaiian bird viruses, they appear to represent different strains of avipoxvirus. PMID- 10813604 TI - Bat rabies in urban centers in Chile. AB - One hundred and five rabies isolates obtained from domestic animals and insectivorous bats in Chile between 1977 and 1998 were molecularly characterized by limited sequence analysis of their nucleoprotein genes. These isolates were compared with viruses isolated from known domestic and wildlife rabies reservoirs in the Americas to identify potential reservoirs of rabies in Chile. The phylogenetic analyses showed that none of the Chilean isolates segregated with viruses from the terrestrial reservoirs. No non-rabies lyssaviruses were found in this study. The Chilean samples were not related to viruses of the sylvatic cycle maintained by the common vampire bat (Desmodus rotundus) in Latin America. Five genetic variants were identified from insectivorous bats in Chile. The Brazilian free-tailed bat (Tadarida brasiliensis) was identified as the reservoir for the rabies genetic variant most frequently isolated in the country between 1977 and 1998. The close association of a group of viruses obtained from a domestic dog (Canis familiaris), Brazilian free-tailed bats, and a red bat (Lasiurus borealis) with viruses maintained by Lasiurus spp. in North America implicated species of this genus as the possible reservoirs of this particular genetic variant in Chile. Reservoirs for the other three variants remain unknown. PMID- 10813605 TI - Association of Moraxella ovis with keratoconjunctivitis in mule deer and moose in Wyoming. AB - Six cases of infectious keratoconjunctivitis (IKC) in mule deer (Odocoileus hemionus) and moose (Alces alces) in Wyoming (USA) were investigated during fall and winter of 1995 and 1996. Excessive lacrimation, mucopurulent conjunctivitis, keratitis, and corneal opacity were observed in mule deer. Moose had severe mucopurulent conjunctivitis, keratitis, and corneal ulceration. Hemolytic, non piliated Moraxella ovis was isolated from two mule deer and two moose. We attempted to reproduce IKC in three mule deer fawns using an isolate of M. ovis from a clinically affected mule deer. These fawns did not develop clinical signs of infection and the bacterium was not reisolated from inoculated deer. Inoculated deer may not have developed clinical signs because deer were not exposed to ultraviolet light or mechanical insult before inoculation. In addition, the isolate used for inoculation may have lost virulence factors through passage, or M. ovis may not have been the primary pathogen responsible for clinical disease in the natural cases of IKC we investigated. The etiology of IKC in free-ranging wild ruminants remains poorly understood. PMID- 10813606 TI - Necrobacillosis in free-living male European bison in Poland. AB - In the Bialowieza Primeval Forest (Poland) a chronic disease of the external genital organs has been observed in free-living male European bison (Bison bonasus) since 1980. Investigations on this disease started in the late 1980s. The most striking findings are necrotic and ulcerative lesions of the prepuce and penis of bison aged from 6 mo to >10 yr. Histologic examination of tissue samples from the prepuce of six bison (9-mo- to 8-yr-old), and from the penis of two bison (3- and 8-yr-old), were characteristic of necrobacillosis. Masses of slender, Gram-negative, rod-like or filamentous bacteria occurred in necrotic tissue. At the periphery of necrotic tissue filamentous bacteria were often arranged in large clusters and strands that advanced towards healthy tissue. Immunolabeling and electron microscopy also suggest that these organisms are Fusobacterium sp. PMID- 10813607 TI - Mycoplasmal conjunctivitis in songbirds from New York. AB - A field study was conducted to determine the prevalence of conjunctivitis and Mycoplasma gallisepticum (MG) infections in house finches (Carpodacus mexicanus) and other songbirds common to bird feeders in Tompkins County (New York, USA). Eight hundred two individuals of 23 species and nine families of birds were captured and given physical examinations during the 14 mo study beginning in February 1998. Clinical conjunctivitis (eyelid or conjunctival swelling, erythema, and discharge) was observed in 10% (19/196) of house finches examined, and only in the winter months from November to March. Unilateral conjunctivitis was observed in 79% (15/19) of affected house finches; one case developed bilateral disease between 8 and 18 days following initial examination. Conjunctivitis was observed in a similar proportion of males and females sampled, and body condition scores and wing chord lengths were not significantly different between diseased and non-diseased house finches. Mycoplasma gallisepticum was isolated from 76% (13/17) of finches with conjunctivitis and 2% (3/168) of clinically normal house finches sampled during the study. DNA fingerprints of 11 MG isolates using random amplification of polymorphic DNA (RAPD) techniques showed no apparent differences in banding patterns over the course of the study, suggesting persistence of a single MG strain in the study population. The prevalence of conjunctivitis and MG infections declined in house finches between February/March 1998 and February/March 1999 (23% to 6%, and 20% to 5%, respectively), but only the former was significant (P < 0.05). Conjunctivitis was also observed in four American goldfinches (Carduelis tristis) and one purple finch (Carpodacus purpureus). Mycoplasma gallisepticum infection was confirmed in the purple finch, the first documented case of MG-associated conjunctivitis in this species. The purple finch isolate was similar to house finch isolates from the study site by RAPD analysis. Positive plate agglutination (PA) tests were recorded in one other goldfinch and two purple finches, suggesting exposure of these individuals to MG. Positive PA tests were also obtained from two brown headed cowbirds (Molothrus ater) and four tufted titmice (Parus bicolor), but MG infection could not be confirmed in these cases due to lack of samples. Based on these findings, the prevalence of MG infections in hosts other than house finches appear to be low in the population sampled. There is growing evidence, however, that songbird species other than house finches are susceptible to MG infection and disease. PMID- 10813608 TI - Immune responses to Mycoplasma conjunctivae in alpine ibex, alpine chamois, and domestic sheep in Switzerland. AB - The humoral immune response of three alpine chamois (Rupicapra rupicapra rupicapra), two alpine ibex (Capra ibex ibex) and three domestic sheep naturally affected with infectious keratoconjunctivitis (IKC), and four ibex and two sheep experimentally infected with Mycoplasma conjunctivae was analysed. In addition, the local immune response to M. conjunctivae was analysed using conjunctival washes from chamois and sheep. Immunoblot analysis of sera using whole cell antigens of M. conjunctivae revealed the major immunogenic proteins which had molecular masses of 175, 83, 68, 60, 50, 42, 36, and 33 kDa. Major antigens were found at 83, 68, 60, and 42 kDa in both sera and conjunctival washes from naturally infected animals of all three Caprinae species. In experimentally infected animals, antibodies to the 68 and 60 kDa antigens were dominant. Naturally infected animals showed much stronger immune reactions than those experimentally infected, and specific antibodies appeared 2 to 4 wk after experimental infection. To evaluate possible cross-reactions, whole cell antigen of M. conjunctivae was analysed by immunoblot against hyperimmune sera of closely related Mycoplasma spp. Antibodies to the 175, 73, 68, 60, and 33 kDa antigens appeared to be specific to M. conjunctivae. Cross-reactions mainly with 83, 50, and 42 kDa antigens were detected, in particular with M. ovipneumoniae and M. bovoculi hyperimmune sera, but also with antisera against M. capricolum capricolum and M. putrefaciens. PMID- 10813609 TI - Pathology of upper respiratory tract disease of gopher tortoises in Florida. AB - Between August 1993 and September 1995, 24 gopher tortoises (Gopherus polyphemus) were received for pathological evaluations from various locations in Florida (USA). All tortoises were examined for clinical signs of upper respiratory tract disease (URTD) including nasal and ocular discharge, palpebral edema, and conjunctivitis. Of the 24 tortoises, 10 had current or previously observed clinical signs of URTD and 14 did not. A blood sample was drawn for detection of anti-mycoplasma antibodies by ELISA, and nasal lavage samples were collected for culture and detection of Mycoplasma agassizii gene sequences by polymerase chain reaction (PCR). Of the 14 clinically healthy tortoises, eight were sero-, culture and PCR-negative, and six were seropositive for antibodies against M. agassizii. Of those six, five were culture- and/or PCR-positive for M. agassizii, and one was culture- and PCR-negative. Of the 10 ill tortoises, nine were seropositive by the ELISA and one was in the suspect range. Nine of the ill tortoises, including the suspect tortoise, were culture- and/or PCR-positive for M. agassizii, and one was culture- and PCR-negative. For histologic evaluation and discussion, the eight sero-, culture-, and PCR-negative tortoises were designated URTD-negative, and the other 16 were classified as URTD-positive. Histologic evaluation of the upper respiratory tract (URT) indicated the presence of mild to severe inflammatory, hyperplastic, or dysplastic changes in 14 URTD-positive tortoises. Seven of eight URTD-negative tortoises had normal appearing nasal cavities; one had mild inflammatory changes. Transmission electron microscopy revealed an organism consistent with Mycoplasma spp. on the nasal mucosal surface of tortoises with clinical signs and lesions of URTD. Additionally, gram-negative bacteria were isolated more frequently from the nasal cavities of URTD-positive tortoises than URTD-negative tortoises. Because clinical signs of URTD were never observed in six of the URTD-positive tortoises, we also conclude that subclinical URTD can occur in gopher tortoises. PMID- 10813610 TI - Retrospective and longitudinal study of salmonellosis in captive wildlife in Trinidad. AB - Morbidity and mortality of captive wildlife at the Emperor Valley Zoo, Trinidad from 1993 to 1996 were analysed to determine involvement of Salmonella spp. A 6 mo longitudinal study was conducted to determine the frequency of isolation of Salmonella spp. from apparently healthy, sick and dead wild mammals, birds, and reptiles. The antibiograms of Salmonella isolates were determined using the disc diffusion method. Fecal samples randomly selected from animal enclosures and cloacal swabs of snakes were cultured for Salmonella spp. following enrichment in tetrathionate and selenite cystine broths. For the 1993-96 period, Salmonella spp. was implicated in 17 (12%) of 141 sick or dead animals and the predominant serotype was S. typhimurium. During the 6 mo prospective study in a mean animal population of 1,186, there were 20 (2%) and 14 (1%) animals that were sick and died respectively; Salmonella spp. was implicated in only one mortality. Overall, of 1,012 samples from apparently healthy wildlife cultured, 66 (7%) yielded 24 serotypes of Salmonella. The predominant serotype were S. seigburg (16 isolates), S. gaminara (6 isolates), and S. thompson (6 isolates). None of the samples yielded S. typhimurium. The frequency of isolation of Salmonella spp. in reptiles (14%) was significantly higher than found in either mammals (7%) or birds (3%). Sixty-five (99%) of 66 Salmonella spp. isolates exhibited resistance to one or more of the nine antimicrobial agents tested. Resistance was high to cephalothin (92%), moderate to streptomycin (35%) and tetracycline (29%), but significantly low to gentamicin (2%), chloramphenicol (0%), and sulphamethoxazole/trimethoprim (0%). The prevalence of asymptomatic infections by Salmonella spp. in zoo animals was high and the very high prevalence of antimicrobial resistance could be a problem when treating salmonellosis. PMID- 10813611 TI - Biochemical and hematologic reference intervals for free-ranging desert bighorn sheep. AB - Over 200 clinically normal desert bighorn sheep (Ovis canadensis) from multiple geographic areas were sampled utilizing a uniform protocol. The goals of this study were to develop comprehensive reference intervals for hematologic and biochemical analytes using central 90th percentile nonparametric analyses. Adult female sheep had greater erythrocyte mass (hemoglobin and hematocrit) compared with adult male sheep. Young animals < or = 1-yr-old had greater erythrocyte mass (hemoglobin, hematocrit and red blood cell count), higher alkaline phosphatase activity, and lower serum protein and globulin concentrations compared with adult animals. Because of the large sample size, wide geographic range, and uniform sample and handling protocol in this study, these reference intervals should be robust and applicable to other free-ranging desert bighorn sheep populations. PMID- 10813612 TI - Trace mineral and vitamin concentrations in the liver and serum of wild muskoxen from Victoria Island. AB - Selected trace minerals and vitamins were assayed in the liver and serum of 25 wild muskoxen (Ovibos moschatus) from Victoria Island, (Nunavut, Canada) in November, 1995. Mean +/- SE liver concentrations in micromol/kg wet weight were 260+/-16 for copper; 1.04+/-0.06 for selenium; 11.5+/-0.7 for molybdenum and 62.8+/-3.3 for vitamin E. Mean +/- SE serum concentrations in micromol/L were 14.2+/-0.3 for copper; 0.75+/-0.04 for selenium, 1.53+/-0.07 for vitamin A and 5.80+/-0.55 for vitamin E. Comparison of liver and serum concentrations of copper, selenium and vitamin E showed that the concentration in one tissue was a relatively poor indicator of the concentration in the other. The copper molybdenum interaction often seen in domestic species was not observed. In general, the concentrations of metals and vitamins found in muskoxen were comparable to those in other ungulates although serum vitamin E concentrations were about one-fourth of those expected. PMID- 10813613 TI - Risk factors associated with capture-related death in eastern wild turkey hens. AB - Capture-related mortality has been a notable risk in the handling of eastern wild turkey (Meleagris gallopavo silvestris). Our objective was to evaluate how environmental factors influence risk and identify physiological correlates that could be used to identify susceptible birds. During winter (January-March) 1995 97, 130 eastern wild turkey hens were captured in southeastern Oklahoma and radiocollared. Of those, 20 hens died < or =14 days of capture. Serum creatine kinase activity (CK; P < 0.01), body temperature (P < 0.01), processing time (P = 0.02), and ambient temperature (P < 0.01) showed a positive relationship with mortality that occurred within 14 days of capture. Plasma corticosterone concentration (P = 0.08) and relative humidity (P < 0.01) showed a negative relationship with mortalities that occurred within 14 days post-capture. Stepwise logistic regression selected CK activity, relative humidity, and ambient temperature as the best predictors of mortality within 14 days post-capture. Our data suggest that susceptible individuals may be identified from CK activity and that capture-related mortality may be minimized by establishing guidelines of when to curtail capture operations based on various weather conditions. PMID- 10813614 TI - Serosurvey for selected infectious disease agents in free-ranging black and white rhinoceros in Africa. AB - Two hundred and eighty one serum samples collected from free-ranging black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros, in the Republic of South Africa (RSA), Namibia, and Kenya from 1987-97, were examined for antibody to 16 different infectious agents. Positive antibody titers were detected against Akabane (59.8%), bluetongue (55%), African horse sickness (27.9%), epizootic haemorrhagic disease of deer (19.4%), parainfluenza type 3 (25.3%), bovine herpes virus 1 (3.1%), equine herpes virus 1 (8.8%) and bovine viral diarrhea (1.2%) viruses, and four serovars of Leptospira interrogans, (ranging 1.2 to 8.8%). No antibody was detected against Rift Valley fever virus, encephalomyocarditis virus, Brucella abortus, and Trypanosoma equiperdum. Interspecies differences were detected for African horse sickness, epizootic haemorrhagic disease of deer and parainfluenza type 3 viruses. There appeared to be some geographic variation in the prevalence of antibody for African horse sickness, bluetongue, epizootic haemorrhagic disease of deer, parainfluenza type 3, equine herpes virus 1 and Leptospira interrogans serovar bratislava. PMID- 10813615 TI - Evaluation of isoflurane and propofol anesthesia for intraabdominal transmitter placement in nesting female canvasback ducks. AB - Heart rate, occurrence of apnea, body temperature, quality of anesthesia and nest abandonment were compared during either propofol or isoflurane anesthesia of nesting female canvasback ducks (Aythya valisineria) at 15 to 18 days of incubation. One hundred eighteen canvasbacks were assigned randomly to three treatments so that nest abandonment could be compared among treatments from May to July 1995 and 1996. Sterile dummy silicone implants were placed during an abdominal laparotomy while ducks were anesthetized with either propofol or isoflurane, or ducks were flushed from the nest but not captured (control). Propofol was delivered through an intravenous catheter, while isoflurane was delivered in oxygen. Propofol provided smooth, rapid induction and recovery, whereas ducks recovering from isoflurane tended to struggle. At the nest, ducks in the propofol group were given additional boluses until they were lightly anesthetized, whereas birds that received isoflurane were released. All birds survived surgery but one death occurred prior to surgery in 1995 using propofol during a period without ventilation and monitoring. Adequate artificial ventilation is recommended to prevent complications. Heart rate declined significantly in both years during isoflurane anesthesia and in 1995 during propofol anesthesia but not 1996. During both isoflurane and propofol anesthesia, body temperature declined significantly over time. Nest abandonment was significantly different among treatments and occurred in all treatment groups in both years, but propofol (15%) and control groups (8%) had lower than expected abandonment compared to isoflurane (28%). Propofol offers several advantages over isoflurane for field use; equipment is easily portable, lower anesthetic cost, and ambient temperature does not alter physical characteristics of the drug. Advantages over isoflurane, including lower nest abandonment following intraabdominal radio transmitter placement, make propofol a good anesthetic choice for field studies. PMID- 10813617 TI - Post-mortem investigations on stranded dolphins and porpoises from Hong Kong waters. AB - Stranded cetaceans reported from the territorial waters of Hong Kong during the period May 1993 to March 1998 were examined to establish factors that may have contributed to their death. During the current study, 28 Indo-Pacific hump-backed dolphins (Sousa chinensis), 32 finless porpoises (Neophocaena phocaenoides), and four bottlenose dolphins (Tursiops truncatus) were necropsied. Bacteria (15 species) were isolated from nine animals. Of these bacteria, 47% were of possible fecal origin reflecting the high level of sewage contamination in Hong Kong's waters. One finless porpoise displayed wounds caused by a shark attack, and two female finless porpoises presented prolapsed uteri. At least 10 finless porpoises showed evidence of moderate to heavy lungworm infections (Halocercus pingi), and this appears to have been a factor contributing to death in at least six animals. Evidence suggesting blunt traumatic injury (probably caused by boat collisions) was found in six cetaceans (three finless porpoises and three hump-backed dolphins). Signs of fishery-related mortality were detected in at least nine animals (six hump-backed dolphins, two finless porpoises, and one bottlenose dolphin). Of these two human-caused mortality types, pre-existing disease or bacterial infection were detected in 29% of cases. Results indicate that human factors may have played a significant role in the death of at least 15 animals (32% of hump-backed dolphins, 15% of finless porpoises, and 25% of bottlenose dolphins). PMID- 10813616 TI - Butorphanol/xylazine/ketamine immobilization of free-ranging Baird's tapirs in Costa Rica. AB - Cardiopulmonary effects and the utility of a butorphanol/xylazine/ketamine combination were evaluated during twenty immobilizations of sixteen Baird's tapirs (Tapirus bairdii) between March 1996 and January of 1998 in Corcovado National Park (Costa Rica). The animals were attracted to a bait site and darted from tree platforms. The tapirs were estimated to weigh between 200 to 300 kg. Actual weights of three tapirs taken at later dates fell within the estimated range. A butorphanol, 48+/-1.84 (x +/- SE) mg/animal IM, and xylazine, 101+/-2.72 mg/animal IM, combination was used to immobilize the animals. In some instances, ketamine was used either IM or IV at 187+/-40.86 mg/animal to prolong the immobilization period in addition to the butorphanol/xylazine combination. Naltrexone was used IM to reverse butorphanol at 257+/-16.19 mg/animal. Either yohimbine, 34+/-0.61 or tolazoline at 12+/-10.27 mg/animal, was used to reverse xylazine. The mean time from dart impact to first visible effect was 4.63+/-0.50 min (x +/- SE). Mean time to sternal recumbency was 12.21+/-1.08 min. Mean time the tapirs were immobilized was 45.63+/-3.6 min. Mean time to return to sternal recumbency and standing in animals that received yohimbine and naltrexone was 3.16+/-1.06 and 5.33+/-1.45 min, respectively. Mean time to return to sternal recumbency and standing in animals that received tolazoline and naltrexone was 1.57+/-0.39 and 3.14+/-0.51 min, respectively. Cardiopulmonary parameters including heart rate, respiratory rate, body temperature, electrocardiogram, percent oxygen satoration, and indirect blood pressure were recorded. Arterial blood gas analysis was performed on four animals. A mild degree of hypoxemia was evidenced by low arterial oxygen saturations. Five of 14 (36%) animals measured had oxygen saturations below 90%. Bradycardia (heart rates <45 BPM) was an expected finding in 11 (55%) immobilizations. Induction, recovery and muscle relaxation of each immobilization was graded. Premature arousal, which occurred in six (30%) animals, was the only problem associated with the immobilizations. Butorphanol/xylazine is a recommended protocol for immobilization of calm, free ranging tapirs lasting less than 30 min. Supplemental intravenous administration of ketamine is recommended for longer procedures. Nasal insufflation of oxygen is recommended. PMID- 10813618 TI - Acute sarcocystosis in a captive white-tailed deer in Virginia. AB - A 19-mo-old female captive white-tailed deer in a public wild animal park in Richmond (Virginia, USA) was necropsied and evaluated histologically following spontaneous death after a 1 wk period of lethargy in a captive herd of 22 deer. An acute necrotizing pneumonia was associated with intraendothelial protozoal schizonts that were identified immunohistochemically as Sarcocystis spp. This is the first confirmed report of acute sarcocystosis in a wild ruminant. PMID- 10813619 TI - Ultrastructure of Frenkelia sp. from a Norwegian lemming in Finland. AB - An apparently healthy Norwegian lemming (Lemmus lemmus) caught in northern Finland was observed to have a whitish body 0.5 to 1.0 mm in diameter in the external layer of the cerebral cortex. By light microscopy a highly lobulated cyst of Frenkelia sp. was observed. By transmission electron microscopy lemmus) collected in the cyst wall was seen consisting of a parasitophorous vacuolar membrane, an underlying electron-dense layer and a granular layer. The membrane was only slightly convoluted. The protrusions of the cyst wall appeared round but were often not distinctive. A very thin septum divided the interior of the cyst into compartments packed with bradyzoites and maturing zoites. The bradyzoites were elongate measuring 5-8 x 1.5-2 microm. This is the first electron microscopical study of Frenkelia sp. from L. lemmus. PMID- 10813620 TI - Pulmonary protostrongyliasis in a mountain hare from Italy. AB - An adult male mountain hare (Lepus timidus), shot during the hunting season on the slopes of Mt. Civetta (eastern Alps, Veneto, northeastern Italy), was eviscerated and diffuse nodular lesions of verminous pneumonia were observed. Adult helminths were collected and male specimens were identified as Protostrongylus pulmonalis. Histologically, a non-purulent interstitial pneumonia and a desquamative bronchiolitis and alveolitis, along with thickening of alveolar septa, were apparent in the nodules. In several enlarged alveoli, a large number of eggs and larvae of nematodes were found. This is the first recorded case of protostrongyliasis in mountain hares from Italy and from an alpine population. PMID- 10813621 TI - Meningeal worm in deer from western Nebraska. AB - One hundred seventy-eight white-tailed deer (Odocoileus virginianus) and 275 mule deer (Odocoileus hemionus) collected from locker plants in the western 2/3 of Nebraska (USA) in November 1997 were examined for the meningeal worm (Parelaphostrongylus tenuis). Parelaphostrongylus tenuis was identified in 17 (10%) of 168 white-tailed deer and in one (<1%) of 273 mule deer. This is the first naturally occurring infection of P. tenuis recorded in a mule deer. PMID- 10813622 TI - Serologic detection of adenoviral hemorrhagic disease in black-tailed deer in California. AB - An enzyme-linked immunosorbent assay (ELISA) and a serum neutralization (SN) test were developed to measure serum antibodies against the adenovirus causing hemorrhagic disease in free-ranging and captive experimentally-infected black tailed deer (Odocoilenus hemionus columbianus) in California (USA). There was a strong (rho = 0.874) and significant (P < 0.0001) correlation between ELISA and SN titers, although the SN assay was more sensitive than the ELISA. PMID- 10813623 TI - Proliferative pododermatitis associated with virus-like particles in a northern gannet. AB - Small multifocal lesions of proliferative pododermatitis were observed in an emaciated adult male northern gannet (Morus bassanus). Ultrastructurally, these lesions were associated with numerous virus-like particles with a size and morphology suggestive of Papovaviridae. DNA in situ hybridization with probes for avian polyomaviral and papillomaviral nucleic acid and an immunohistochemical test for the presence of papillomaviral antigen failed to identify this virus further. To our knowledge, papovavirus-like particles have not been recognized previously in this avian species. PMID- 10813625 TI - New records of sylvatic plague in Kansas. AB - Sylvatic plague, or plague of wild rodents is caused by Yersinia pestis and entered California (USA) from Asia about 1899. Extensive sampling during the 1930's and 1940's documented the spread of plague to approximately its current distribution in North America. Records from the Centers for Disease Control and Prevention document plague in Kansas (USA) between 1945 and 1950, but since then there has been no documentation of plague in the state. Following a die-off of a black-tailed prairie dog (Cynomys ludovicianus) colony on the Cimarron National Grassland, in the southwestern corner of Kansas (3710'N, 10145'W), we sampled fleas from burrows in June 1997, and tested them for Yersinia pestis. Twelve of 13 pools of Oropsyla hirsuta and one of two Pulex sp. were positive. A similar sample of fleas, from another colony where black-tailed prairie dogs were active at the time, yielded no positive fleas. PMID- 10813624 TI - Septicemic pasteurellosis in free-ranging neonatal pronghorn in Oregon. AB - As part of a study to determine the cause(s) of population decline and low survival of pronghorn (Antilocapra americana) neonates on Hart Mountain National Antelope Refuge (HMNAR), Oregon (USA), 55 of 104 neonates captured during May 1996 and 1997 were necropsied (n = 28, 1996; n = 27, 1997) to determine cause of death. Necropsies were conducted on fawns that died during May, June, or July of each year. The objectives of this study were to report the occurrence and pathology of pasteurellosis in neonates and determine if the isolated strain of Pasteurella multocida was unique. Septicemic pasteurellosis, caused by P. multocida, was diagnosed as the cause of death for two neonates in May and June 1997. Necropsy findings included widely scattered petechial and ecchymotic hemorrhages found over a large portion of the subcutaneous tissue, meninges of the brain, epicardium, skeletal muscle, and serosal surface of the thorasic and abdominal cavities. Histological examination of lung tissues revealed diffuse congestion and edema and moderate to marked multifocal infiltrate of macrophages, neutrophils, and numerous bacteria within many terminal bronchioles and alveoli. Pasteurella multocida serotypes A:3,4, and B:1 were isolated from several tissues including lung, intestinal, thorasic fluid, and heart blood. Each B:1 isolate had DNA restriction endonuclease fingerprint profiles distinct from isolates previously characterized from domestic cattle, swan (Olor spp.), moose (Alces alces), and pronghorn from Montana (USA). This is the first report of pasteurellosis in pronghorn from Oregon and the B:1 isolates appear to be unique in comparison to DNA fingerprint profiles from selected domestic and wild species. PMID- 10813626 TI - Normal hematologic and serum biochemical reference intervals for juvenile wild turkeys. AB - Blood samples taken from 48 4-mo-old wild turkeys (Meleagris gallopova silvestris) were used to establish reference intervals for hematology and serum chemistry values. The study was conducted during September and October 1996. Packed cell volume, total and differential white cell counts, total protein, albumin, glucose, calcium, uric acid, triglyceride concentrations, as well as aspartate transaminase (AST) and lactate dehydrogenase (LDH) activities were assayed. Reference intervals from wild turkeys are similar to those reported for domestic turkeys. PMID- 10813627 TI - Hematology and serum chemistry of the island fox on Santa Cruz Island. AB - Serum and hematologic biochemistry values for island foxes (Urocyon littoralis) on Santa Cruz Island (California, USA) in April (wet season) and September (dry season) 1998 were evaluated. Serum chemistry of island foxes generally varied seasonally; 10 (40%) of the 25 serum characteristics were higher in the wet season, and three (12%) of the 25 serum characteristics were higher in the dry season. No hematologic parameters varied between seasons, although some measures varied between sexes. Blood analytes also varied with age; fox pups had higher values than adults for one hematologic and four serum parameters, whereas adult foxes had higher values for five hematologic characteristics. The information on blood chemistry provides baseline data useful in the monitoring of this threatened insular endemic carnivore. PMID- 10813628 TI - Mammary gland tumors in captive African hedgehogs. AB - From December 1995 to July 1999, eight mammary gland tumors were diagnosed in eight adult captive female African hedgehogs (Atelerix albiventris). The tumors presented as single or multiple subcutaneous masses along the cranial or caudal abdomen that varied in size for each hedgehog. Histologically, seven of eight (88%) mammary gland tumors were malignant. Tumors were classified as solid (4 cases), tubular (2 cases), and papillary (2 cases). Seven tumors had infiltrated into the surrounding stroma and three tumors had histologic evidence of neoplastic vascular invasion. Three hedgehogs had concurrent neoplasms. These are believed to be the first reported cases of mammary gland tumors in African hedgehogs. PMID- 10813629 TI - The handling of polar bears in the western Hudson Bay. PMID- 10813630 TI - Pharmacological clues to calmodulin-mediated activation of skeletal ryanodine receptor using [3H]-ryanodine binding. AB - The hypothesis that calmodulin (CaM) may act as a positive modulator of junctional SR Ca2+-release channel/ ryanodine receptor (RyRl) rests largerly on the demonstrated capacity of CaM to interact structurally and functionally with RyRl at pCa > 8 (Tripathy et al., 1995). The goal of the present [3H]-ryanodine binding study was to produce, in isolated terminal cisternae (TC) and in purified junctional face membrane (JFM), CaM-mediated activation of RyRl at less extreme pCa values, i.e. closer to resting myoplasmic pCa, and to analyze more accurately the corresponding changes in binding affinity for ryanodine of the receptor. We were able to monitor these changes at an optimum pCa of 6.5, following pre activation of native RyRl by mM concentrations of caffeine or microM concentrations of antraquinone compound doxorubicin, and at various doses of these triggers. CaM increased the affinity of ryanodine binding to isolated TC in the presence of 1 mM AMP-PCP as an activator of RyRl; the Kd for ryanodine binding was reduced from 21.8 nM to 13.2 nM by 1microM CaM. Similar effects of CaM were seen when AMP-PCP was replaced by either caffeine or doxorubicin. In order to discount the involvement of SR extrajunctional proteins in this effect, the experiments were repeated on purified JFM. Again, CaM increased the affinity of ryanodine binding; the Kd was reduced from 11.1 nM to 7.0 nM by 1 microM CaM (in the presence of doxorubicin). Pharmacological triggers of CaM-activatory action on native RyRl, like caffeine and doxorubicin, have long been characterized for their ability to activate RyRl by increasing the Ca2+ sensitivity of the receptor. We speculate that the triggering effect of these agents on the CaM-mediated mechanism in vitro might mimick one of the early effects of the activation of RyRl in skeletal muscle, during E-C coupling. PMID- 10813631 TI - Force-velocity and unloaded shortening velocity during graded potassium contractures in frog skeletal muscle fibres. AB - Steady-state conditions of contraction, at maximal and submaximal forces, were produced in intact single muscle fibres, from Rana esculenta, using full tetani and graded K+-contractures. The uniformity in radial direction. of spreading of activation produced in K+-contractures, was checked in relation to the fibre diameters. The absolute isometric force was similar in tetani and maximal contractures, for fibres with diameters between 40 and 60 microm, but not for fibres with diameters greater than about 70 microm in which contracture force never reached tetanic force. The force [K+]o relation was similar for fibres with diameters between 40 and 60 microm. but it was right shifted and it had a minor slope for fibres with diameters greater than 65-70 microm. This suggests that only in the small diameter fibres (40-60 microm) the activation does not fail to penetrate uniformly from the surface towards the fibre core. For fibres selected in the diameter range between 40 and 60 microm, force-velocity relations and unloaded shortening velocities were determined in tetani and maximal and submaximal contractures. Data were obtained across a force range of 0.3 to 1 P0 (tetanic plateau force). Controlled velocity method was used to obtain force velocity relations, and slack test to determine the unloaded shortening velocity (VU). The values of the parameters characterising the force velocity relation (V0 and a/P0) and VU as determined by the slack test did not differ significantly in tetani and contractures, independent of the activation level or absolute force developed by the fibre. These results show that. at least within the range of forces tested. crossbridge kinetics is independent of the number of cycling crossbridges, in agreement with the prediction of the 'recruitment' model of myofilament activation. PMID- 10813632 TI - Measured and modeled properties of mammalian skeletal muscle: III. the effects of stimulus frequency on stretch-induced force enhancement and shortening-induced force depression. AB - Stretch-induced force enhancement and shortening-induced force depression were examined in fast-twitch feline caudofemoralis muscle at 37 degrees C. These phenomena were induced by applying ramp length changes during the first 100--200 ms of an otherwise isometric contraction. The effects of various stimulus frequencies ranging from 30 to 120 pps were investigated over lengths ranging from 0.85 to 1.15 L0. Distributed asynchronous stimulation of bundles of ventral roots was employed to produce smooth contractions at sub-tetanic stimulus frequencies in whole muscle. Of the two components of force enhancement identified by Noble (1992) we observed only the transient component that decays with time; we did not observe residual force enhancement. The force depression that we observed was symmetrical in almost all respects to the transient force enhancement, and was unlike the shortening-induced de-activation and residual force depression identified by Edman (Edman. 1975; Edman et al., 1993). Both transient force enhancement and depression were independent of work, load and activation. Reversals in the direction of ramp length changes following either an initial stretch or initial shortening were shown to cancel the effects of both transient force enhancement and transient force depression. The distances over which these cancellations could be achieved were different for the lengthening and shortening effects. This asymmetry can be reconciled with the predictions of Huxley's original cross-bridge mechanism by incorporating the recent suggestion that myosin heads can interact with multiple actin binding sites during a single 'working' stroke. We conclude that the types of force enhancement/ depression that are most likely to be encountered under physiological conditions are the transient effects observed here, but that even these will have relatively little effect on force production during most natural behaviors. PMID- 10813633 TI - Measured and modeled properties of mammalian skeletal muscle: IV. dynamics of activation and deactivation. AB - The interactive effects of length and stimulus frequency on rise and fall times and on sag were investigated in fast-twitch feline caudofemoralis at normal body temperature. The length and stimulus frequency ranges studied were 0.8 1.2 L0 and 15 60 pps. Isometric rise times were shortest under two sets of conditions: short lengths + low stimulus frequencies and long lengths + high stimulus frequencies. In contrast the isometric fall time relationship showed a single minimum at short lengths + low stimulus frequencies. Velocity was shown to have an additional effect on fall time, but only at higher stimulus frequencies (40 60 pps): fall times were shorter during movement in either direction as compared to isometric. The effects of sag were greatest at shorter lengths and lower stimulus frequencies during isometric stimulus trains. Potential mechanisms underlying this last effect were investigated by comparing isometric twitches elicited prior to and immediately following a sag-inducing stimulus train. Post-sag twitches produced less force, reached peak force earlier and initially decayed more quickly compared to pre-sag twitches. However, the final rate of force decay and the initial rate of force rise (during the first 15 ms) were unaffected by sag. We construct a logical argument based on these findings to hypothesize that the predominant mechanism underlying sag is an increase in the rate of sarcoplasmic calcium ion removal. All of the above findings were used to construct a model of activation dynamics for fast-twitch muscle, which was then extrapolated to slow twitch muscle. When coupled with a previous model of kinematic dynamics, the complete model produced accurate predictions of the forces actually recorded during experiments in which we applied concurrent dynamic changes in length. velocity and stimulus frequency. PMID- 10813634 TI - Expression of cofilin isoforms during development of mouse striated muscles. AB - Cofilin (CF) is an actin regulatory protein that plays a critical role in actin filament dynamics in a variety of cells. Two cofilin isoforms. muscle-type (M-CF) and nonmuscle-type (NM-CF) encoded by different genes, exist in mammals; in the adult, the former is predominantly expressed in muscle tissues, while the latter is distributed in various non-muscle tissues (Ono et al., 1994). In this study, we examined cofilin isoform expression during skeletal and cardiac muscle development in mice using cDNA probes and antibodies which distinguish the isoforms. We found that the expression of M-CF was initiated in terminally differentiated myogenic cells in both the myotome and limb buds. In myogenic cell cultures, its expression occurred coupled with myotube formation. NM-CF was expressed in developing skeletal and cardiac muscles but disappeared from skeletal muscle during postnatal development, while its expression persisted in the heart, even in adult mice. A similar situation was observed in the heart of other mammals. Thus, it is likely that the both cofilin isoforms are involved in the regulation of actin assembly during myofibrillogenesis. Only M-CF could be involved in actin dynamics in mature skeletal muscle, while both isoforms could be in the mature heart. PMID- 10813635 TI - The length dependency of calcium activated contractions in the femoral artery smooth muscle studied with different methods of skinning. AB - The relationship between the calcium concentration and the isometric tension obtained with different techniques of skinning provides information on the biochemical events of contraction in vascular smooth muscle. Muscle preparations of the rabbit femoral artery were skinned with triton X-100, saponin, beta-escin and alpha-toxin and the relationship between the calcium concentration and isometric tension was determined at different preparation lengths. We determined the calcium sensitivity as a function of muscle length with different techniques of skinning. At a pCa of 6.0, triton X-100 skinned smooth muscle of the femoral artery generated 50% of the maximal tension. In alpha-toxin skinned preparations, this calcium sensitivity was shifted to a pCa of 5.6. The sensitivity of the saponin and 3-escin skinned preparations were in between those of the triton X 100 and the alpha-toxin skinned preparations. The cooperativity of the regulation of contraction varied among the differently skinned preparations between 3 (alpha toxin) and 6 (triton X-100). The relationships between the calcium concentration and the isometric tension of the differently skinned preparations up to the optimal length for tension generation did not exhibit any length dependency. The length tension relationship, obtained from the maximal response at the highest calcium concentration is in line with that from other studies. The presence of intracellular proteins and membranes affects the regulation of contraction in smooth muscle of the femoral artery. PMID- 10813636 TI - The spectrin skeleton of newly-invaginated plasma membrane. AB - As a cell's shape and volume change. its surface area must re-adjust. How is the plasma membrane's spectrin skeleton implicated? For erythrocytes, cells of fixed surface area, spectrin responses to mechanical disturbances have been studied, but for more typical cells with changeable surface areas, they have not. In rapidly shrinking cells, surface membrane at an adherent substratum invaginates, forming transient vacuole-like dilations (VLDs). We exploited this readily inducible surface area perturbation to pose a simple question: is newly invaginated plasma membrane naked or is it supported by a spectrin skeleton? The spectrin skeleton was examined immunocytochemically in L6 cells (rat skeletal muscle) before and after VLD formation, using fixation in cold methanol and 4112, an antibody against beta-fodrin and beta-spectrin. 4112 was visualized by confocal fluorescence microscopy, while paired phase contrast images independently located the VLDs. To generate VLDs, cells were hypotonically swelled then reshrunk in isotonic medium. Swollen L6 cells maintained their plasma membrane (sarcolemma) spectrin skeleton. Within minutes of subsequent shrinkage, VLDs of 1-2 microm diameter invaginated at the substratum surface of myotubes. Both sarcolemma and VLDs were lined by a relatively uniform spectrin skeleton. Z-series suggested that some of the spectrin skeleton-lined sarcolemma became internalized as vacuoles. PMID- 10813639 TI - Do cross-bridges contribute to the tension during stretch of passive muscle? PMID- 10813638 TI - Activation of locomotor and grasping spine muscle fibres in chaetognaths: a curious paradox. AB - Chaetognath muscle fibres resemble vertebrate muscle fibres in having an abundant sarcoplasmic reticulum (SR) and analogues of the transverse (T) tubular system. but contraction is regulated differently. In intact chaetognaths electrically evoked contractions of the striated locomotor muscles were largely or totally blocked by d-tubocurarine, by surgical removal of the ventral ganglion and by Co2 +. Contractions of single cells enzymatically dissociated from locomotor muscles were likewise blocked by Co2+, they twitched once only after calciseptine, showed neither contractures nor elevated intracellular Ca2+ with caffeine, and ryanodine did not block contractions. Whole cell voltage-clamped locomotor muscle cells displayed a typical inward rectified Ca2 + current that was sensitive to the Ca2+ channel blockers nifedipine and calciseptine and showed voltage-dependent activation with a threshold at approximately-25 mV and a peak inward current at approximately + 10 mV. In contrast, whole cell voltage-clamped cells from the muscles operating the grasping spines of the head showed an initial very rapid and rapidly-inactivating inward current abolished by tetrodotoxin (TTX), followed by a slower and slowly-inactivating inward current blocked by calciseptine. The relation between these observations and the unusual 'vertebrate-like' structure of the muscle cells is discussed. PMID- 10813637 TI - Loop diuretics inhibit detubulation and vacuolation in amphibian muscle fibres exposed to osmotic shock. AB - The effect of loop diuretics at concentrations known to influence cellular water entry coupled to Na-K-Cl co-transport, upon the vacuolation and detubulation following osmotic shock, was investigated in amphibian skeletal muscles. These were exposed to a glycerol-Ringer solution (18 min), an isotonic Ca2+/Mg2+ Ringer solution and cooling. Adding bumetanide (1.0 and 2.0 microM) to these solutions sharply reduced the incidence of detubulation, assessed by abolition or otherwise of action potential after-depolarisations, from 93.9 +/- 4.7% (n = 6) to 5.0 +/- 1.1% (n = 4: mean +/- SEM: 2.0 microM bumetanide). It dramatically reduced the number and fraction of muscle volume occupied by tubular vacuoles, measured using confocal microscopy, from 60.3 +/- 4.3% (n = 10) to 9.0 +/- 1.1% (n = 35). The incidence of large horseradish peroxidase-lined tubular vacuoles, viewed using electronmicroscopy, similarly was reduced with 2 microM bumetanide in the glycerol-Ringer solution. Bumetanide acted through cellular volume adjustments early in the detubulation protocol. Thus, it exerted its maximum effect when added to the glycerol-Ringer, rather than the Ca2+/Mg2+ Ringer solution. Furthermore, whereas fibre diameters measured using scanning electron microscopy returned to normal during glycerol treatment relative to those of control fibres left in isotonic Ringer, addition of 2.0 microM bumetanide in the glycerol Ringer left markedly smaller fibre diameters. Finally equipotent concentrations of the chemically distinct loop diuretics. furosemide and ethacrynic acid similarly influenced detubulation. These findings implicate Na-K-Cl co-transport in the water entry into muscle fibres that would be expected following introduction of extracellular glycerol. This might then enable the subsequent Na-K-ATPase dependent water extrusion that produces the tubular distension (vacuolation) and detachment (detubulation) following glycerol withdrawal, phenomena also observed in muscular dystrophy. PMID- 10813640 TI - Consonant enhancement: promising findings. PMID- 10813641 TI - Consonant enhancement effects on speech recognition of hearing-impaired children. AB - Differences in gain (enhancement, in dB) required to optimize the consonant/vowel intensity ratio in nonsense syllables were determined for stops and fricatives, both voiced and voiceless, in 12 children with congenital moderate to severe sensorineural hearing loss. The test stimuli were vowel/consonant nonsense syllables with various levels of enhancement ranging from 0 dB (for the unprocessed stimulus) to 24 dB of gain, in steps of 3 or 6 dB. Results showed that significant improvements in consonant recognition can be obtained with individualized adjustment of consonant amplitude for children as young as 5 years of age. PMID- 10813642 TI - Comparison of air-conduction and bone-conduction hearing thresholds for pure tones and octave-band filtered sound effects. AB - The purpose of this study was to measure air-conduction (AC) and bone-conduction (BC) hearing thresholds with pure-tone and filtered sound effect stimuli using standard audiometric equipment. A group of 20 young, normal-hearing listeners participated in the study. Pure-tone stimuli were 250, 500, 1000, 2000, and 4000 Hz. Sound effect stimuli were 12 natural sounds that were spectrally limited to an octave bandwidth centered at either 250, 500, 1000, 2000, or 4000 Hz. The AC and BC detection thresholds were measured using a clinical audiometer (Madsen Orbiter 922) with a B-71 bone oscillator and TDH-50 earphones. Results indicated that detection thresholds for the pure-tone and corresponding octave-band sound effect stimuli were within 3 to 4 dB of each other for both AC and BC testing. The findings support the notion that octave-filtered sound effects are a viable alternative to pure-tone stimuli for use in audiology clinics. PMID- 10813643 TI - Word recognition performance for Northwestern University Auditory Test No. 6 word lists in quiet and in competing message. AB - Word recognition norms were determined for the Northwestern University Auditory Test No. 6 (NU-6) released on the Department of Veterans Affairs (VA) Disc 1.1. Word recognition performance (in quiet and in competing message) was measured at 12 presentation levels for 24 young adults with normal hearing. Test-retest reliability also was evaluated. Word recognition scores for VA Disc 1.1 were generally higher than those reported for VA Disc 1.0. The differences in NU-6 scores across disc versions, however, were small and unlikely to affect clinical decisions based on word recognition tests. Score differences on test-retest for the VA Disc 1.1 version of the NU-6 with a competing message background also were small and unlikely to affect clinical outcomes. Overall, based on comparisons of scores for the disc versions of the NU-6, it appears that the two different recordings can be used interchangeably for clinical applications. PMID- 10813644 TI - Otoacoustic emission findings in Rett syndrome. AB - Rett syndrome is a neurologic disorder affecting mainly females after a seemingly normal 6 to 18 months of life. The resulting developmental disabilities include apparent dementia and loss of acquired language, social skills, and purposeful hand use. The present investigation assessed 10 individuals with Rett syndrome and a control group matched for age and sex. The present study aimed to determine the clinical feasibility of obtaining otoacoustic emissions (OAEs) from the Rett syndrome group and to compare the characteristics of the transient evoked and distortion-product OAEs obtained from the two subject groups. Results indicated that OAE evaluation is a clinically feasible method of assessing individuals with Rett syndrome. The Rett syndrome group had less robust OAEs, especially in the higher frequencies, when compared to the control group. Seven of the Rett syndrome group were identified as having reduced or absent OAEs in at least one ear. These findings suggest a need for hearing screening at an early age and monitoring of hearing on a regular basis. PMID- 10813645 TI - Comparison of performance with wide dynamic range compression and linear amplification. AB - This study compared subject performance and preference using a compression limiting hearing aid set to linear amplification (program 1) and wide dynamic range compression (WDRC, program 2). The frequency responses of the hearing aid were matched to a 65 dB SPL signal and maximum output to a 90 dB SPL signal. Twenty subjects with moderate to moderately severe sensorineural hearing loss were tested. Speech recognition scores and speech reception thresholds were obtained both in quiet and in noise. Subjective preference for WDRC or linear amplification was measured via a paired-comparison procedure on "loudness appropriateness," "clarity," and "pleasantness" to continuous discourse presented in quiet and in noise. Results suggested that WDRC yielded better speech intelligibility in quiet for low-level signals and no difference in speech intelligibility in noise compared to linear amplification. Subjects preferred WDRC for loudness to both high- and low-level signals and for pleasantness to high-level signals. PMID- 10813646 TI - Variables affecting the use of prescriptive formulae to fit modern nonlinear hearing aids. AB - It is routine for audiologists to fit nonlinear hearing aids by using one of several current prescriptive formulae. While such an approach might result in acceptable fittings for single-channel analog compression hearing aids, its direct application to fitting current multichannel compression hearing aids may result in less than an optimal fit. In this paper, the effects of filter bandwidth, number of channels, detector type, compression threshold, attack and release times, and distortion on the final output of a nonlinear compression hearing aid are discussed. The subjective results of such effects and its implications on our clinical practice will be explored as well. PMID- 10813647 TI - Chemiluminescent aldehyde and beta-diketone reactions promoted by peroxynitrite. AB - Peroxynitrite is shown here to promote the aerobic oxidation of isobutanal (IBAL) and 3-methyl-2,4-pentanedione (MP) in a pH 7.2 phosphate buffer into acetone plus formate and biacetyl plus acetate, respectively. These products are expected from dioxetane intermediates, whose thermolysis is known to be chemiluminescent (CL). Accordingly, the extent of total oxygen uptake by IBAL at different concentrations parallels the corresponding CL maximum intensities. The pH profile based on oxygen uptake data for the MP reaction matches the titration curve of peroxynitrous acid (pK(a) approximately 7), indicating that peroxynitrite anion is the oxidizing agent. Energy transfer studies with IBAL and the 9, 10 dibromoanthracene-2-sulfonate ion, a triplet carbonyl detector, indicates that triplet acetone (tau = 19 micros) is the energy donor. It is postulated that IBAL or MP-generated triplet carbonyls are produced by the thermolysis of dioxetane intermediates, which are formed by the cyclization of alpha-hydroperoxide intermediates produced by insertion of dioxygen into the IBAL or MP enolyl radicals, followed by their reduction. Accordingly, EPR spin-trapping studies with 3,5-dibromo-4-nitrosobenzenesulfonic acid (DBNBS) and 2-methyl-2 nitrosopropane (MNP) revealed the intermediacy of carbon-centered radicals, as expected for one-electron abstraction from the enol forms of IBAL or MP by peroxynitrite. The EPR data obtained with IBAL also reveal formation of the isopropyl radical produced by competitive nucleophilic addition of ONOO(-) to IBAL, followed by homolytic cleavage of this adduct and beta-scission of the resulting Me(2)CHCH(O(-))O(*). Superstoichiometric formation of fragmentation products from IBAL or MP attests to the prevalence of an autoxidation chain reaction, here proposed to be initiated by one-electron abstraction by ONOO(-) from the substrate. This work reveals the potential role of peroxynitrite as a generator of electronically excited species that may contribute to deleterious and pathological processes associated with excessive nitric oxide and aldehyde production. PMID- 10813648 TI - Acylation of protein lysines by trichloroethylene oxide. AB - Stable lysine adducts were formed in proteins following reaction with trichloroethylene (TCE) oxide, the major reactive compound generated by the metabolism of TCE. The order of formation of these adducts is N(6)-formyllysine > N(6)-(dichloroacetyl)lysine >> N(6)-glyoxyllysine, with the ratio being influenced by the particular protein. Protein lysine adducts were also analyzed following the enzymatic oxidation of TCE with several different cytochrome P450 (P450) enzyme systems. The ratio of formyl/dichloroacetyl lysine adducts was influenced by the enzyme system that was used. Chloral and TCE oxide formation was more extensive with rat liver microsomes isolated from phenobarbital-treated rats than with rat microsomes in which P450 2E1 was induced by treatment with isoniazid or in human P450 2E1 systems. Glutathione (GSH) and GSH transferase had inhibitory effects on the reaction of TCE oxide with albumin, with formylation being atteunated much more than the formation of dichloroacetyllysine. GSH is likely to react with the reactive acyl chloride intermediates formed from TCE oxide hydrolysis, instead of direct reaction with TCE oxide, as judged by the lack of an effect of GSH on the rate of decomposition of TCE oxide. Studies with the model enzymes aldolase and glucose-6-phosphate dehydrogenase, both known to have sensitive lysine groups, indicate that TCE oxide has effects similar to known acylating agents that form the same adducts; concentrations of TCE oxide (or the model acylating agents) in the low-millimolar range were needed for inhibition. The characterization of TCE-derived protein adducts can be used as a basis for consideration of the exposure and risk of TCE to humans. Human P450 2E1 was less likely to oxidize TCE to form TCE oxide and protein lysine adducts than rat P450 2B1, and the difference is rationalized in terms of the influence of the protein on chloride migration in an enzyme reaction intermediate. PMID- 10813649 TI - Studies on the in vivo biotransformation of the tobacco alkaloid beta-nicotyrine. AB - This paper reports the results of studies on the in vivo metabolic fate of the tobacco alkaloid 1-methyl-2-(3-pyridinyl)pyrrole (beta-nicotyrine) in New Zealand white rabbits. Two previously characterized metabolites, 5-hydroxy-1-methyl-5-(3 pyridinyl)-2-pyrrolidinone (5-hydroxycotinine) and 2-hydroxy-1-methyl-5-(3 pyridinyl)-3-pyrrolin-2-one, were present in low concentrations in the urine of the treated animals. The major urinary metabolite of beta-nicotyrine was identified as cis-3'-hydroxy-1-methyl-5-(3-pyridinyl)-2-pyrrolidinone (cis-3' hydroxycotinine), the diastereoisomer of the major urinary metabolite of (S) nicotine. The pathway leading to cis-3'-hydroxycotinine is proposed to proceed via autoxidation of 2-hydroxy-1-methyl-5-(3-pyridinyl)pyrrole, a postulated cytochrome P450-generated metabolite of beta-nicotyrine, followed by reduction of the carbon-carbon double bond present in the resulting 3-hydroxy-3-pyrrolin-2-one species. This proposal is supported by the in vivo biotransformation of 2-acetoxy 1-methyl-5-(3-pyridinyl)pyrrole, a latent form of the putative hydroxypyrrole intermediate, to cis-3'-hydroxycotinine. The in vivo conversion of 5-hydroxy-1 methyl-5-(3-pyridinyl)-3-pyrrolin-2-one to 5-hydroxycotinine is offered as evidence that supports the proposed reduction step. PMID- 10813650 TI - A metabolite of equine estrogens, 4-hydroxyequilenin, induces DNA damage and apoptosis in breast cancer cell lines. AB - Estrogen replacement therapy has been correlated with an increased risk of developing breast or endometrial cancer. 4-Hydroxyequilenin (4-OHEN) is a catechol metabolite of equilenin which is a minor component of the estrogen replacement formulation marketed under the name of Premarin (Wyeth-Ayerst). Previously, we showed that 4-OHEN autoxidizes to quinoids which can consume reducing equivalents and molecular oxygen, are potent cytotoxins, and cause a variety of damage to DNA, including formation of bulky stable adducts, apurinic sites, and oxidation of the phosphate-sugar backbone and purine/pyrimidine bases [Bolton, J. L., Pisha, E., Zhang, F., and Qiu, S. (1998) Chem. Res. Toxicol. 11, 1113-1127]. All of these deleterious effects could contribute to the cytotoxic and genotoxic effects of equilenin in vivo. In the study presented here, we examined the relative toxicity of 4-OHEN in estrogen receptor (ER) positive cells (MCF-7 and S30) compared to that in breast cancer cells without the estrogen receptor (MDA-MB-231). The data showed that 4-OHEN was 4-fold more toxic to MCF-7 cells (LC(50) = 6.0 +/- 0. 2 microM) and 6-fold more toxic to S30 cells (LC(50) = 4.0 +/- 0.1 microM) than to MDA-MB-231 cells (LC(50) = 24 +/- 0.3 microM). Using the single-cell gel electrophoresis assay (comet assay) to assess DNA damage, we found that 4-OHEN causes concentration-dependent DNA single-strand cleavage in all three cell lines, and this effect could be enhanced by agents which catalyze redox cycling (NADH) or deplete cellular GSH (diethyl maleate). In addition, the ER(+) cell lines (MCF-7 and S30) were considerably more sensitive to induction of DNA damage by 4-OHEN than the ER(-) cells (MDA-MB-231). 4-OHEN also caused a concentration-dependent increase in the amount of mutagenic lesion 8-oxo-dG in the S30 cells as determined by LC/MS-MS. Cell morphology assays showed that 4 OHEN induces apoptosis in these cell lines. As observed with the toxicity assay and the comet assay, the ER(+) cells were more sensitive to induction of apoptosis by 4-OHEN than MDA-MB-231 cells. Finally, the endogenous catechol estrogen metabolite 4-hydroxyestrone (4-OHE) was considerably less effective at inducing DNA damage and apoptosis in breast cancer cell lines than 4-OHEN. Our data suggest that the cytotoxic effects of 4-OHEN may be related to its ability to induce DNA damage and apoptosis in hormone sensitive cells in vivo, and these effects may be potentiated by the estrogen receptor. PMID- 10813651 TI - A novel approach for analyzing the structure of DNA modified by Benzo[a]pyrene diol epoxide at single-molecule resolution. AB - Benzo[a]pyrene diol epoxide (BPDE) has been shown to bind specifically to the exocyclic amino group of deoxyguanosine in duplex DNA. Interestingly, this metabolite exhibits stereoselectivity in its tumorigenic and mutagenic effects. It is thought that local DNA conformation is altered at the site of the adduct, resulting in aberrant biological processes, and that in certain sequence contexts BPDE-DNA adducts induce bends in the DNA. In the work presented here, we compared DNA structural alterations of BPDE-modified DNA and unmodified DNA via tapping mode atomic force microscopy (AFM). DNA fragments 366 base pairs (bp) in length were generated by PCR from the duplicated multiple-cloning site of pBEND2 inserted into pGEM-3Zf(-), and either mock-modified or treated with BPDE to give modification levels between 1 and 5% of the nucleotides. Control or BPDE-modified DNA was adsorbed to mica and visualized in air by AFM. The contour lengths and end-to-end lengths of individual molecules were measured. The ratio of end-to-end distance to contour length was significantly smaller for modified DNA molecules than for the unmodified DNA preparation, although the frequency distributions of the contour lengths were similar for the two preparations. This suggests BPDE-DNA adducts cause significant bending of DNA molecules, confirming previous conclusions based on more indirect measurements. The average induced bend angle for BPDE-DNA adducts is estimated to be at least 30 degrees. PMID- 10813652 TI - Identification of novel small molecule ligands that bind to tetanus toxin. AB - Tetanus toxin belongs to a family of clostridial protein neurotoxins for which there are no known antidotes. Another closely related member of this family, botulinum toxin, is being used with increasing frequency by physicians to treat severe muscle disorders. Botulinum toxin has also been produced in large quantities by terrorists for use as a biological weapon. To identify small molecule ligands that might bind to the targeting domain of tetanus and botulinum toxins and to facilitate the design of inhibitors and new reagents for their detection, molecular docking calculations were used to screen a large database of compounds for their potential to bind to the C fragment of tetanus toxin. Eleven of the predicted ligands were assayed by electrospray ionization mass spectrometry (ESI-MS) for binding to the tetanus toxin C fragment, and five ligands (45%) were found to bind to the protein. One of these compounds, doxorubicin, was observed to have strong hydrophobic interactions with the C fragment. To check the ligands for their ability to compete with ganglioside binding, each was also tested using a GT1b liposome assay. Doxorubicin was the only ligand found to competitively bind the tetanus toxin C fragment with an appreciable binding constant (9.4 microM). PMID- 10813653 TI - Electrophilic properties of patulin. Adduct structures and reaction pathways with 4-bromothiophenol and other model nucleophiles. AB - The mycotoxin patulin (PAT) is believed to exert its cytotoxic and chromosome damaging effects by forming covalent adducts with essential cellular thiols. Since the chemical structures of such adducts are unknown to date, we have studied the reaction of PAT and its O-acetylated derivative with the monofunctional thiol model compound 4-bromothiophenol (BTP), which was chosen due to analytical advantages. By means of analytical and preparative high-performance liquid chromatography, 16 adducts of PAT and 3 adducts of acetyl-PAT were isolated and their chemical structures elucidated by (1)H and (13)C NMR, IR, and UV spectroscopy. Time course studies and analysis of daughter product formation from isolated intermediate adducts led to a detailed scheme for the reaction of PAT with BTP. The structures of adducts of PAT formed with other model nucleophiles, e. g., the aliphatic thiol 2-mercaptoethanol and the aromatic amine 4-bromoaniline, were also elucidated and found to corroborate the reaction scheme. In addition, one further reaction pathway was observed with 2 mercaptoethanol, which appears to be independent from those found for BTP. Our study with model nucleophiles provides insights into the electrophilic reactivity of PAT and proved to be useful for the structure elucidation of PAT adducts with biological nucleophiles of toxicological relevance, as will be reported by Fliege and Metzler [(2000) Chem. Res. Toxicol. 13, 373-381]. PMID- 10813654 TI - Electrophilic properties of patulin. N-acetylcysteine and glutathione adducts. AB - In our studies on the electrophilic properties of the mycotoxin patulin (PAT), we have now investigated the nonenzymatic reaction of PAT with the thiol-containing tripeptide glutathione and its metabolic degradation product N-acetyl-L-cysteine (NAC). Adduct formation in aqueous phosphate buffer (pH 7.4) was studied by analytical HPLC/DAD, and most of the products were isolated by preparative HPLC. Structure elucidation was carried out mainly by means of high-resolution NMR experiments and comparison of the data with those previously obtained for PAT adducts formed with simple model nucleophiles such as 4-bromothiophenol and 2 mercaptoethanol [Fliege, R., and Metzler, M. (2000) Chem. Res. Toxicol. 13, 363 372]. The assigned structures were confirmed by UV spectroscopy, formation of daughter products from isolated adducts, and partly FAB-MS. The reaction pathways of PAT with NAC were qualitatively the same as those previously observed for the aliphatic thiol model compound 2-mercaptoethanol. Due to the chiral nature of NAC and the new chiral center generated during the reaction with PAT, two diastereomers of each adduct were formed and observed in HPLC analysis. The major products formed in the reaction of PAT with GSH were of the same structural type as obtained with NAC. In addition, three cyclic adducts were formed with GSH, arising from the nucleophilic activity of the alpha-amino groups of the glutamic acid and the cysteine residue. In contrast, free cysteine yielded a markedly different adduct pattern, possibly due to the preferred formation of mixed thiol/amine-type adducts involving the alpha-amino group. PMID- 10813656 TI - A novel nitration product formed during the reaction of peroxynitrite with 2',3',5'-tri-O-acetyl-7,8-dihydro-8-oxoguanosine: N-nitro-N'-[1-(2,3,5-tri-O acetyl-beta-D-erythro-pentofuranosyl)- 2, 4-dioxoimidazolidin-5 ylidene]guanidine. AB - A novel nitration product, formed during the reaction of peroxynitrite with 2',3',5'-tri-O-acetyl-7,8-dihydro-8-oxoguanosine, has been characterized using a combination of UV/vis, CD, and NMR spectroscopy and mass spectrometry. This compound has been identified as N-nitro-N'-[1-(2,3, 5-tri-O-acetyl-beta-D-erythro pentofuranosyl)-2, 4-dioxoimidazolidin-5-ylidene]guanidine (IV). Upon base hydrolysis, IV releases nitroguanidine (IVa) and an intermediate, 1-(2,3, 5-tri-O acetyl-beta-D-erythro-pentofuranosyl)-5-iminoimidazolidine -2, 4-dione (IVb). This intermediate is ultimately hydrolyzed to the stable 3-(2,3,5-tri-O-acetyl beta-D-erythro-pentofuranosyl)oxaluric acid (IVc). IV can be reduced by sodium borohydride to a pair of stable diastereomers (IV(red)()). The formation of this product is rationalized in terms of initial oxidation of 2',3', 5'-tri-O-acetyl 7,8-dihydro-8-oxoguanosine to a quinonoid diimine intermediate, 3. Nucleophilic attack at C5 of 3 by peroxynitrite leads to formation of a C5-oxyl radical species, 5, which then undergoes a series of rearrangements to yield an ylidene radical, 7. Combination of this radical species with nitrogen dioxide results in the formation of product IV. PMID- 10813655 TI - Mobilization of iron from coal fly ash was dependent upon the particle size and source of coal: analysis of rates and mechanisms. AB - The observed iron mobilization rate from size-fractionated coal fly ash is consistent with the model predictions for a limiting case of mass transfer where the dominant resistance is diffusion through a layer of depleted solid between the surface of spherical particles and a shrinking core of unreacted material. The rate of mobilization of iron from coal fly ash under physiologically relevant conditions in vitro was previously shown to depend on the size of the ash particles and on the source of the coal, and these in vitro measurements have been shown to correlate with indirect measurements of excess iron in cultured cells. Existing iron mobilization data were compared to mathematical models for mass transfer and chemical reaction in solid-liquid heterogeneous systems. Liquid phase diffusion resistance can be ruled out as the rate-limiting mechanism for iron mobilization as the model predictions for this case are clearly inconsistent with the measurements. Other plausible hypotheses, such as a rate limited by a heterogeneous surface reaction, cannot be conclusively ruled out by the available data. These mathematical analysis methods are applicable to the design of future experiments to determine the rate-limiting mechanism for the mobilization of iron and of other transition metals from both ambient air samples and surrogates for major sources of particulate air pollution. PMID- 10813657 TI - Novel 1,N(6)-etheno-2'-deoxyadenosine adducts from lipid peroxidation products. AB - trans,trans-2,4-Decadienal (DDE) is a widespread alpha, beta-unsaturated aldehyde found, for example, in food, water, and environmental pollutants. DDE is also endogenously generated as a breakdown product of lipid peroxidation in cell membranes. In the work presented here, the reaction of DDE with 2'-deoxyadenosine (dAdo) was investigated in an effort to assess its possible DNA damage potential. Besides 1,N(6)-etheno-2'-deoxyadenosine and two products, namely, 1-[3-(2-deoxy beta-D-erythro-pentofuranosyl)-3H-imidazo[2, 1-i]purin-7-yl]-1,2,3-octanetriol (adduct I) and 1-[3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3H-imidazo[2, 1 i]purin-7-yl]-1,2-heptanediol (adduct II), previously described by our group, two novel etheno adducts were identified. Thus, 1-[3-(2-deoxy-beta-D-erythro pentofuranosyl)-3H-imidazo[2, 1-i]purin-7-yl]-1-hexanol (adduct III) and 1-[3-(2 deoxy-beta-D-erythro-pentofuranosyl)-3H-imidazo[2, 1-i]purin-7-yl]-2,3-epoxy-1 octanol (adduct IV) were isolated by reverse-phase high-performance liquid chromatography and characterized on the basis of extensive spectroscopic measurements. The formation of the adducts is likely to involve initial DDE oxidation followed by generation of reactive intermediates such as diepoxides, epoxides, and/or hydroperoxides. The subsequent reaction of the latter oxidation products with dAdo will give rise to the four described adducts. We also demonstrated here that upon oxidation, DDE reacts with calf thymus DNA, producing the four dAdo adducts. Interestingly, two of them are the expected products arising from the reaction of dAdo with 4-hydroxy-trans-2-nonenal (HNE) and trans 2-octenal, two other important breakdown lipid peroxidation products. The reactivity of DDE with DNA is lower than that of the latter aldehydes. However, DDE produced a wider variety of adducts. The characterization of the different DNA-etheno adducts and the determination of the mechanism of formation are of great importance for a better understanding of the deleterious biological effects associated with this class of compounds. PMID- 10813658 TI - Polyclonal antibodies to a fluorescent 4-hydroxy-2-nonenal (HNE)-derived lysine lysine cross-link: characterization and application to HNE-treated protein and in vitro oxidized low-density lipoprotein. AB - Oxidative modification of low-density lipoprotein (LDL) is thought to play a key role in the etiology of atherosclerosis. Oxidized LDL that accumulates in atherosclerotic plaques is known to exhibit a characteristic fluorescence with excitation and emission near 360 and 430 nm, respectively. (E)-4-Hydroxy-2 nonenal (HNE), formed during LDL oxidation, is capable of modifying LDL to generate the same fluorescent signature. The HNE-derived fluorophore was shown by us to possess a 2-hydroxy-2-pentyl-1,2-dihydropyrrol-3-one iminium (HPDPI) structure. We herein report the synthesis of the HPDPI-derived lysine-lysine cross-link needed as a standard reference for HPLC quantitation of the cross-link in protein hydrolysates. The main focus of the current work, however, is the design and development of two polyclonal antibodies against the HPDPI epitope. Utilizing these antibodies, levels of the HPDPI epitope were estimated in HNE treated bovine serum albumin and in copper-oxidized LDL by an enzyme-linked immunosorbent assay. Our results are consistent with the premise that the fluorescent HPDPI cross-link is a key contributor to the fluorescence exhibited by atherosclerotic lesions. PMID- 10813659 TI - Doxorubicin metabolism and toxicity in human myocardium: role of cytoplasmic deglycosidation and carbonyl reduction. AB - The anthracycline doxorubicin (DOX) is an exceptionally good antineoplastic agent, but its use is limited by formation of metabolites which induce acute and chronic cardiac toxicities. Whereas the acute toxicity is mild, the chronic toxicity can produce a life-threatening cardiomyopathy. Studies in laboratory animals are of limited value in predicting the structure and reactivity of toxic metabolites in humans; therefore, we used an ethically acceptable system which is suitable for exploring DOX metabolism in human myocardium. The system involves cytosolic fractions from myocardial samples obtained during aorto-coronary bypass grafting. After reconstitution with NADPH and DOX, these fractions generate the alcohol metabolite doxorubicinol (DOXol) as well as DOX deoxyaglycone and DOXol hydroxyaglycone, reflecting reduction of the side chain carbonyl group, reductase type deglycosidation of the anthracycline, and hydrolase-type deglycosidation followed by carbonyl reduction, respectively. The efficiency of each metabolic route has been evaluated at low and high DOX:protein ratios, reproducing acute, single-dose and chronic, multiple-dose regimens, respectively. Low DOX:protein ratios increase the efficiency of formation of DOX deoxyaglycone and DOXol hydroxyaglycone but decrease that of DOXol. Conversely, high DOX:protein ratios facilitate the formation of DOXol but impair reductase- or hydrolase-type deglycosidation and uncouple hydrolysis from carbonyl reduction, making DOXol accumulate at levels higher than those of DOX deoxyaglycone and DOXol hydroxyaglycone. Structure-activity considerations have suggested that aglycones and DOXol may inflict cardiac damage by inducing oxidative stress or by perturbing iron homeostasis, respectively. Having characterized the influence of DOX:protein ratios on deglycosidation or carbonyl reduction, we propose that the benign acute toxicity should be attributed to the oxidant activity of aglycones, whereas the life-threatening chronic toxicity should be attributed to alterations of iron homeostasis by DOXol. This picture rationalizes the limited protective efficacy of antioxidants against chronic cardiomyopathy vis-a-vis the better protection offered by iron chelators, and forms the basis for developing analogues which produce less DOXol. PMID- 10813661 TI - Mutagenic potential of guanine N(2) adducts of butadiene mono- and diolepoxide volume 13, number 1, january 2000, pp 18-25 PMID- 10813662 TI - The Complete Zeldovich Approximation. AB - We have developed a generalization of the Zeldovich approximation that is exact in a wide variety of situations, including planar, spherical, and cylindrical symmetries. We have shown that this generalization, which we call the complete Zeldovich approximation (CZA), is exact to second order at an arbitrary point within any field. For Gaussian fields, the third-order error has been obtained and shown to be very small. For statistical purposes, the CZA leads to results exact to the third order. PMID- 10813660 TI - Synthesis of nucleosides and oligonucleotides containing adducts of acrolein and vinyl chloride. AB - Vinyl chloride and acrolein are important industrial chemicals. Both form DNA adducts, vinyl chloride after enzymatic oxidation to chlorooxirane and acrolein by direct reaction. Reaction at the N(2) position of guanine is a major pathway. The resulting 2-oxoethyl and 3-oxopropyl adducts cyclize spontaneously to hydroxyethano and hydroxypropano derivatives, respectively. The two cyclic adducts have been detected in DNA exposed to these mutagens. A new method has been developed for the synthesis of deoxyguanosine adducts of chlorooxirane and acrolein, as well as oligonucleotides containing these adducts. Reaction of O(6) [(trimethylsilyl)ethyl]-2-fluoro-2'-deoxyinosine with the appropriate aminodiol followed by oxidative cleavage of the diol with NaIO(4) gave the adducts in excellent yields. Reaction of oligonucleotides containing the halonucleoside with the aminodiols followed by NaIO(4) efficiently created the nucleosides in the oligonucleotides. Deoxyadenosine adducts were created similarly using 6 chloropurine 9-(2'-deoxyriboside). PMID- 10813663 TI - Simulating the Effects of Intergalactic Gray Dust. AB - Using a high-resolution cosmological hydrodynamic simulation, we present a method to constrain extinction due to intergalactic gray dust based on the observed magnitudes of distant Type Ia supernovae. We apply several simple prescriptions to relate the intergalactic dust density to the gas density in the simulation, thereby obtaining dust extinctions that may be directly compared with the observed distribution of supernova magnitudes. Our analysis is sensitive to the spatial distribution of gray dust but is not dependent on its intrinsic properties, such as its opacity or grain size. We present an application of our technique to the supernova data of Perlmutter et al., who find that their high redshift sample is approximately 0.2 mag fainter than the expectation for a nonaccelerating, low-density universe. We find that for gray dust to be responsible, it must be distributed quite smoothly (e.g., tracing intergalactic gas). More realistic dust distributions, such as dust tracing the metal density, are inconsistent with observations at the 1.5-2 sigma level. Upcoming observations and improved modeling of the dust distribution should lead to stronger constraints on intergalactic gray dust extinction. PMID- 10813664 TI - Fluid Dark Matter. AB - Dark matter modeled as a classical scalar field that interacts only with gravity and with itself by a potential that is close to quartic at large field values and approaches a quadratic form when the field is small would be gravitationally produced by inflation and, at the present epoch, could act like an ideal fluid with pressure that is a function only of the mass density. This could have observationally interesting effects on the core radii and solid-body rotation of dark matter halos and on the low-mass end of the primeval mass fluctuation power spectrum. PMID- 10813665 TI - Weak Gravitational Lensing by the Nearby Cluster Abell 3667. AB - We present two weak lensing reconstructions of the nearby (zcl=0.055) merging cluster Abell 3667, based on observations taken approximately 1 yr apart under different seeing conditions. This is the lowest redshift cluster with a weak lensing mass reconstruction to date. The reproducibility of features in the two mass maps demonstrates that weak lensing studies of low-redshift clusters are feasible. These data constitute the first results from an X-ray luminosity selected weak lensing survey of 19 low-redshift (z<0.1) southern clusters. PMID- 10813666 TI - Chandra X-Ray Observations of the Hydra A Cluster: An Interaction between the Radio Source and the X-Ray-emitting Gas. AB - We present Chandra X-ray observations of the Hydra A cluster of galaxies, and we report the discovery of structure in the central 80 kpc of the cluster's X-ray emitting gas. The most remarkable structures are depressions in the X-ray surface brightness, approximately 25-35 kpc in diameter, that are coincident with Hydra A's radio lobes. The depressions are nearly devoid of X-ray-emitting gas, and there is no evidence for shock-heated gas surrounding the radio lobes. We suggest that the gas within the surface brightness depressions was displaced as the radio lobes expanded subsonically, leaving cavities in the hot atmosphere. The gas temperature declines from 4 keV at 70 kpc to 3 keV in the inner 20 kpc of the brightest cluster galaxy (BCG), and the cooling time of the gas is approximately 600 Myr in the inner 10 kpc. These properties are consistent with the presence of an approximately 34 M middle dot in circle yr-1 cooling flow within a 70 kpc radius. Bright X-ray emission is present in the BCG surrounding a recently accreted disk of nebular emission and young stars. The star formation rate is commensurate with the cooling rate of the hot gas within the volume of the disk, although the sink for the material that may be cooling at larger radii remains elusive. A bright, unresolved X-ray source is present in the BCG's nucleus, coincident with the radio core. Its X-ray spectrum is consistent with a power law absorbed by a foreground NH approximately 4x1022 cm-2 column of hydrogen. This column is roughly consistent with the hydrogen column seen in absorption toward the less, similar24 pc diameter VLBA radio source. Apart from the point source, no evidence for excess X-ray absorption above the Galactic column is found. PMID- 10813667 TI - Mitigating Charge Transfer Inefficiency in the Chandra X-Ray Observatory Advanced CCD Imaging Spectrometer. AB - The ACIS front-illuminated CCDs on board the Chandra X-Ray Observatory were damaged in the extreme environment of the Earth's radiation belts, resulting in enhanced charge transfer inefficiency (CTI). This produces a row dependence in gain, event grade, and energy resolution. We model the CTI as a function of input photon energy, including the effects of detrapping (charge trailing), shielding within an event (charge in the leading pixels of the 3x3 event island protects the rest of the island by filling traps), and nonuniform spatial distribution of traps. This technique cannot fully recover the degraded energy resolution, but it reduces the position dependence of gain and grade distributions. By correcting the grade distributions as well as the event amplitudes, we can improve the instrument's quantum efficiency. We outline our model for CTI correction and discuss how the corrector can improve astrophysical results derived from ACIS data. PMID- 10813668 TI - The Structure and Evolution of Weakly Self-interacting Cold Dark Matter Halos. AB - The evolution of halos consisting of weakly self-interacting dark matter particles is investigated using a new numerical Monte Carlo N-body method. The halos initially contain kinematically cold, dense r-1 power-law cores. For interaction cross sections sigma*=sigmawsi&solm0;mp>/=10-100 cm2 g-1, weak self interaction leads to the formation of isothermal, constant-density cores within a Hubble time as a result of heat transfer into the cold inner regions. This core structure is in good agreement with the observations of dark matter rotation curves in dwarf galaxies. The isothermal core radii and core densities are a function of the halo scale radii and scale masses which depend on the cosmological model. Adopting the currently popular LambdaCDM model, the predicted core radii and core densities are in good agreement with the observations. For large interaction cross sections, massive dark halos with scale radii rs>/=1.4x104 cm2 g-1 (sigma*)-1 kpc could experience core collapse during their lifetime, leading to cores with singular isothermal density profiles. PMID- 10813669 TI - The Late Afterglow and Host Galaxy of GRB 990712. AB - We present deep Hubble Space Telescope (HST) imaging, as well as ground-based imaging and spectroscopy, of the optical afterglow associated with the long duration gamma-ray burst GRB 990712 and its host galaxy. The data were obtained 48-123 days after the burst occurred. The magnitudes of the host (R=21.9, V=22.5) and optical afterglow (R=25.4, V=25.8, 47.7 days after the burst) favor a scenario in which the optical light follows a pure power-law decay with an index of alpha approximately -1.0. We find no evidence for a contribution from a supernova like SN 1998bw. This suggests that either there are multiple classes of long-duration gamma-ray bursts or that the peak luminosity of the supernova was more than 1.5 mag fainter than SN 1998bw. The HST images and EFOSC2 spectra indicate that the gamma-ray burst was located in a bright, extended feature (possibly a star-forming region) 1.4 kpc from the nucleus of a 0.2L*B galaxy at z=0.434, possibly a Seyfert 2 galaxy. The late-time afterglow and host galaxy of GRB 990712 bear some resemblance to those of GRB 970508. PMID- 10813670 TI - Dust Echoes from Gamma-Ray Bursts. AB - The deviation from the power-law decline of the optical flux observed in GRB 970228 and GRB 980326 has been used recently to argue in favor of the connection between gamma-ray bursts and supernovae. We consider an alternative explanation for this phenomenon, based on the scattering of a prompt optical burst by 0.1 M middle dot in circle dust located beyond its sublimation radius 0.1-1 pc from the burst. In both cases, the optical energy observed at the time of the first detection of the afterglow suffices to produce an echo after approximately 20-30 days, as observed. Prompt optical monitoring of future bursts and multiband photometry of the afterglows will enable us to test simple models of dust reprocessing quantitatively and to predict source redshift. PMID- 10813671 TI - Flash Heating of Circumstellar Clouds by Gamma-Ray Bursts. AB - The blast-wave model for gamma-ray bursts (GRBs) has been called into question by observations of spectra from GRBs that are harder than can be produced through optically thin synchrotron emission. If GRBs originate from the collapse of massive stars, then circumstellar clouds near burst sources will be illuminated by intense gamma radiation, and the electrons in these clouds will be rapidly scattered to energies as large as several hundred keV. Low-energy photons that subsequently pass through the hot plasma will be scattered to higher energies, hardening the intrinsic spectrum. This effect resolves the "line-of-death" objection to the synchrotron shock model. Illuminated clouds near GRBs will form relativistic plasmas containing large numbers of electron-positron pairs that can be detected within approximately 1-2 days of the explosion before expanding and dissipating. Localized regions of pair annihilation radiation in the Galaxy would reveal past GRB explosions. PMID- 10813672 TI - Multiple Sub-Jet Model of Gamma-Ray Bursts and Possible Origin of X-Ray Precursors and Postcursors. AB - We assume that internal shocks of gamma-ray bursts (GRBs) consist of multiple sub jets with a collimation half-angle of about several times gamma-1i, where gammai is the Lorentz factor of each sub-jet. If by chance a sub-jet is first emitted off-axis from the line of sight, the observed peak energy can be in the X-ray region. Next, if by chance a subsequent sub-jet is emitted along the line of sight, then the peak energy will be in the gamma-ray region and the gamma ray may arrive after the X-ray precursor from the former sub-jet depending on parameters. This model predicts a new class of GRBs with extremely weak and short gamma-ray emission but X-ray precursors and/or postcursors as well as an afterglow. PMID- 10813673 TI - The Synchrotron Spectrum of Fast Cooling Electrons Revisited. AB - We discuss the spectrum arising from synchrotron emission by fast cooling (FC) electrons, when fresh electrons are continually accelerated by a strong blast wave, into a power-law distribution of energies. The FC spectrum has so far been described by four power-law segments divided by three break frequencies nusa 75 years (n = 8; age range, 82-93 years) were treated at a reduced dose. Restaging was performed after 3 and 6 cycles, followed by every 3 months for the first 2 years, and every 6 months thereafter. RESULTS: A total of 123 cycles were administered to the study patients, with the median number of 6 cycles per patient (range, 1-9 cycles). At a median follow-up of 24 months (range, 1.5-87+ months), 22 patients were alive without evidence of disease and 3 patients had died (1 patient death was treatment-related). Twenty-four patients who were considered evaluable achieved a complete remission, 21 patients after 3 cycles and the remaining 3 patients after 6 cycles of treatment. Side effects generally were manageable, with only one patient requiring premature discontinuation of treatment due to protracted thrombocytopenia after three courses of therapy, and tolerance was not different between the two age groups. No recurrences were observed at last follow-up. CONCLUSIONS: The authors believe that chemotherapy using the CHOP regimen is highly effective in the treatment of patients with localized primary high grade gastric lymphoma. PMID- 10813709 TI - Combined therapy consisting of intraarterial cisplatin infusion and systemic interferon-alpha for hepatocellular carcinoma patients with major portal vein thrombosis or distant metastasis. AB - BACKGROUND: Hepatocellular carcinoma (HCC) patients with major vascular involvement or extrahepatic metastasis are not good candidates for surgery or transarterial chemoembolization (TACE). In this study, the authors evaluated the efficacy of combined therapy with intraarterial cisplatin infusion and systemic administration of interferon-alpha (IFN-alpha) as a palliative treatment for these patients. METHODS: Sixty-eight HCC patients with major portal vein thrombosis (n = 47) or distant metastasis (n = 27) were randomly allocated to 1 of 3 groups. Group I (n = 19) received combined therapy consisting of intraarterial cisplatin infusion and systemic IFN-alpha, Group II (n = 23) received intraarterial cisplatin infusion, and Group III (n = 26) was managed with only supportive care. Cisplatin 2 mg/kg was infused through the proper hepatic artery every 8 weeks, and IFN-alpha 3 million IU/m2 was administered subcutaneously 3 times a week. RESULTS: The partial response (defined as a 50% or greater reduction in the product of the 2 longest perpendicular tumor measurements) rate of Group I was significantly higher than that of Group II (33% vs. 14%; P < 0.05). Also, the 1-year survival rate of Group I (27%) was higher than that of Group II (9%) or Group III (0%) (P < 0.05 and P < 0.01, respectively). The median survival period of Group I was 19 weeks, which was significantly longer than that of Group II (11 weeks) or Group III (5 weeks) (P < 0.05 and P < 0.01, respectively). CONCLUSIONS: These results suggest that combined therapy consisting of intraarterial cisplatin infusion and systemic IFN alpha may be useful as a palliative treatment for HCC patients with major vascular involvement or extrahepatic metastasis. PMID- 10813710 TI - Medroxyprogesterone acetate inhibits human pancreatic carcinoma cell growth by inducing apoptosis in association with Bcl-2 phosphorylation. AB - BACKGROUND: A previous study found that medroxyprogesterone acetate (MPA) delayed the in vivo growth of three (AsPC-1, Capan-2, and MiaPaCa-2) of nine human pancreatic carcinoma cell lines transplanted into nude mice (Cancer 1995; 75:1263 72). The current study was undertaken to evaluate the basis for this inhibitor. METHODS: The estrogen receptor (ER) and progesterone receptor (PgR) status in nine human pancreatic carcinoma cell lines, AsPC-1, BxPC-3, Capan-1, Capan-2, Hs 700T, Hs-766T, MiaPaCa-2, PANC-1, and SUIT-2, was assessed using an enzyme immunoassay (EIA). The authors tested the growth inhibitory activity of MPA and the morphologic changes in these nine pancreatic carcinoma cell lines. Cell cycle progression and DNA fragmentation also were evaluated in these cell lines. Immunoblot analysis was used to determine bcl-2 expression and phosphorylation. RESULTS: In the EIA assay, ER was detected in three cell lines (BxPC-3, Capan-2, and MiaPaCa-2), and PgR was also detected in three (AsPC-1, Capan-2, and MiaPaCa 2). Medroxyprogesterone acetate inhibited the growth of three cell lines (AsPC-1, Capan-2, and MiaPaCa-2) with IC50 values ranging from 2.3 x 10(7) to 6.1 x 10(-7) M. In these three responsive cell lines, MPA caused cell detachment and decreased cell density. The nuclei of the MPA-treated cells were condensed and often fragmented. Cell cycle analysis of these three cell lines showed that MPA induced the appearance of a sub-G1 peak, which is characteristic of early apoptotic cells. DNA degradation assay after MPA treatment showed a typical DNA ladder pattern consistent with apoptosis. Immunoblot analysis of MPA-treated cells that overexpressed bcl-2 revealed a pattern consistent with bcl-2 phosphorylation. CONCLUSIONS: Clinically attainable concentrations of MPA can inhibit the growth of some human pancreatic carcinoma cells in vitro by inducing apoptosis, probably through their PgR, in association with the phosphorylation of bcl-2. This agent may be useful for treating patients with pancreatic carcinoma. PMID- 10813711 TI - Characterization of a newly established human pancreatic carcinoma cell line, UK Pan-1. AB - BACKGROUND: A highly tumorigenic cell line designated as UK Pan-1 was established in a surgically removed human pancreatic adenocarcinoma and characterized as having many of the genotypic and phenotypic alterations commonly found in pancreatic tumors. METHODS: The cell line was characterized by its morphology, growth rate in monolayer culture and soft agar, tumorigenicity in nude mice, and chromosomal analysis. Furthermore, the status of p53, Ki-ras mutation and transforming growth factor (TGF)-/receptor expression were determined. The characteristics of UK Pan-1 were compared with those of other commonly used pancreatic carcinoma cell lines. RESULTS: Quiescent UK Pan-1 cells could be stimulated to proliferate in growth factor free nutrient media, indicating a growth factor independent phenotype. UK Pan- 1 cells grew in soft agar and rapidly formed tumors in nude mice. This cell line possesses a mutation at codon 12 of the c-Ki-ras-2 gene that is commonly found in pancreatic carcinoma. Fluorescence in situ hybridization showed that two alleles of p53 tumor suppressor gene were present in UK Pan-1. However, sequencing analysis revealed a mutation in one allele at exon 8, codon 273 (G to A; Arg to His). Additional growth assays indicated that the cell line was insensitive to negative growth regulation induced by exogenous TGF-beta. Molecular analysis of the TGF-beta signaling pathway showed that UK Pan-1 did not express appreciable levels of the TGF-beta receptor type I, II, or III mRNAs, but did express DPC4 mRNA. Karyotype analysis revealed an 18q21 deletion indicating a possible loss of heterozygosity for DPC4, as well as other chromosomal deletions and rearrangements. CONCLUSIONS: This study indicates that UK Pan-1 is a highly tumorigenic cell line possessing a molecularly complex pattern of mutations that may be used as a model to further the understanding of the mechanisms responsible for the development of pancreatic carcinoma. PMID- 10813712 TI - Giant cell tumor of the bones of the hand and foot. AB - BACKGROUND: Giant cell tumor of the small bones of the hand and foot is suspected of having some peculiar features compared with giant cell tumor in other sites. Moreover, it could share some features with other giant cell rich lesions involving the hand and foot, and this may affect the differential diagnosis. The aim of this study was to analyze the features of lesions such as these in the files of the Rizzoli Orthopedic Institute. METHODS: The incidence of giant cell tumors of the bones of the hand and foot seen at the Rizzoli Orthopedic Institute over 50 years (1947-1997) was taken into consideration. There were 8 lesions of the hand and 21 of the foot. Clinical information and follow-up of the patients were studied and updated. Radiographs were studied and radiographic features analyzed. Histopathologic material was thoroughly reviewed and histologic features analyzed. RESULTS: Although the location of tumor was helpful information, radiographic features were not specific. Giant cell tumors of the small bones of the hand and foot showed a predominance in females, younger patients and more aggressive behavior than giant cell tumors of large bones. The authors did not observe multicentricity or pulmonary metastases. CONCLUSIONS: Because the radiographic features of giant cell tumor of the hand and foot overlap those of other giant cell rich lesions in these locations, histologic diagnosis is mandatory, although it may be difficult and require the establishment of diagnostic criteria for giant cell tumor. As this tumor tends to be more aggressive than other giant cell rich lesions, treatments of choice are aggressive curettage or resection. PMID- 10813713 TI - Prospective randomized comparison of morning versus night daily single subcutaneous administration of granulocyte-macrophage-colony stimulating factor in patients with soft tissue or bone sarcoma. AB - BACKGROUND: Hematopoietic growth factors (HGFs) have been used to reduce the neutropenic complications of cytotoxic chemotherapy so that higher doses may be given. The authors have previously shown that endogenous serum granulocyte-colony stimulating factor (G-CSF) and granulocyte-macrophage-colony stimulating factor (GM-CSF) levels at night (p.m.) were significantly higher than those in the morning (a.m.). METHODS: Twenty-four patients with soft tissue or bone sarcoma who were treated with high dose ifosfamide-based chemotherapy were enrolled in this study. Patients were randomized to either a.m. or p.m. treatment. GM-CSF was administered at a dose of 5 microg/kg/day at 10 a.m. or 10 p.m., beginning 36-48 hours after the last chemotherapy dose. GM-CSF therapy was continued until the neutrophil count exceeded 1,000/mm3 for 2 consecutive days. Leukocyte, neutrophil, monocyte, and platelet counts were measured immediately before GM-CSF administration and exactly 12 hours after the first dose of GM-CSF, and every 24 hours until 3 days after the cessation of GM-CSF. RESULTS: The mean duration of Grade 3-4 neutropenia was 5.3 +/- 0.4 days for the a.m. treatment arm and 6.5 +/- 0.3 days for the p.m. treatment arm (P = 0.017). Although the duration of neutropenia in the a.m. arm was significantly shorter than in the p.m. arm, there were no differences related to the number of febrile neutropenic episodes or the duration of antibiotic administration. Also, there were no differences in the side effects observed in the a.m. and p.m. arms. CONCLUSIONS: The finding of 1.2 days' difference in the duration of Grade 3-4 neutropenia warrants further study of chronotherapy with HGFs. PMID- 10813714 TI - Treatment of patients with recurrent and primary refractory acute myelogenous leukemia using mitoxantrone and intermediate-dose cytarabine: a pharmacologically based regimen. AB - BACKGROUND: Although chemotherapy can achieve a high rate of disease remission induction in patients with newly diagnosed acute myelogenous leukemia (AML), patients with recurrent or refractory AML generally have a poorer rate of response. This study assessed the utility of mitoxantrone and intermediate-dose cytarabine (Ara-C) in the treatment of patients with recurrent or refractory AML. METHODS: Forty-seven patients with recurrent or refractory AML were treated with Ara-C, 0.5 gm/m2, intravenously (i.v.) every 12 hours x 12 doses on Days 1-6 and mitoxantrone, 5 mg/m2, i.v. on Days 1-5. RESULTS: Twenty-nine of the 47 patients (62%) achieved a complete response. The median duration of disease remission was 112 days (range, 29 days- 8 years). Of the 25 patients age > or = 60 years, 19 (76%) had a complete disease remission and the median duration of disease remission in this group was 114 days (range, 33-370 days), although all patients subsequently developed a disease recurrence. The chemotherapy generally was well tolerated, with a mean duration of neutropenia of 31 days and a mean duration of thrombocytopenia of 33 days. Three patients died of infectious complications between 23-26 days after the initiation of chemotherapy, 1 patient died of sudden cardiac arrest 13 days after the initiation of chemotherapy, and 1 patient developed cutaneous desquamation. Three patients developed acute cerebellar dysfunction. CONCLUSIONS: The use of mitoxantrone and Ara-C is effective in the treatment of patients with recurrent and refractory AML. The subgroup of patients age > or = 60 years also had a high rate of disease remission induction with this regimen, and the regimen generally was well tolerated. PMID- 10813715 TI - High dose inhalation interleukin-2 therapy for lung metastases in patients with malignant melanoma. AB - BACKGROUND: The lungs are a frequent site of metastasis in patients with melanoma, and this may cause respiratory problems in the terminal phase of the illness. Inhalation interleukin (IL)-2 therapy to the lung has been piloted and appears to be well tolerated. METHODS: Twenty-seven patients were treated with single agent dacarbazine and concurrent high dose inhalation IL-2 36 million IU per day). The patients previously had progressed on chemotherapy, predominately dacarbazine-based regimens. Patients included those with American Joint Committee on Cancer Stage IV melanoma, predominately those with lung metastases, but patients with extrapulmonary metastases also were allowed on the study. RESULTS: Five of the 27 patients experienced a complete pulmonary remission. Eight patients achieved a partial pulmonary remission, and 5 patients experienced stabilization of their disease. Eight patients developed pulmonary metastases. One patient was not evaluable. Four of the five patients who achieved a complete response and seven of the eight patients who achieved a partial response previously were treated with dacarbazine and progressed. There were no responses in extrapulmonary metastases. Side effects of treatment were minimal. The complete responses all were durable with a follow-up of 12 months, whereas patients with partial responses and stable disease progressed when IL-2 was discontinued. CONCLUSIONS: Inhalation therapy with IL-2 for pulmonary metastases from melanoma appears to be safe. The current preliminary study suggests efficacy although concurrent chemotherapy was given, thus confounding results to some extent. Therefore, these results need to be reproduced without concomitant chemotherapy. In addition, a strategy comprised of therapy with IL-2 inhalation until disease progression may prolong responses. PMID- 10813716 TI - Effects of high dose raloxifene in selected patients with advanced breast carcinoma. AB - BACKGROUND: An earlier trial of raloxifene, conducted in women with metastatic breast carcinoma who initially had responded to tamoxifen and subsequently developed disease progression, suggested no antitumor activity for raloxifene in tamoxifen-refractory disease. However, preclinical studies and preliminary clinical data in healthy women suggest that raloxifene antagonizes growth of estrogen-dependent neoplasia. METHODS: Raloxifene HCl 150 mg twice daily was given to 22 postmenopausal women with metastatic (American Joint Committee on Cancer Stage IV) or locoregionally recurrent, initially estrogen receptor positive breast carcinoma. Prior systemic treatment of metastatic disease was not allowed. Prior adjuvant chemotherapy or hormonal therapy was required to have been completed at least 1 year before study entry. Tumor response was evaluated every other month either radiographically or by physical examination. Evaluable disease was defined as bidimensionally measurable lesions. RESULTS: Twenty-one patients were eligible for efficacy analysis; 6 had been treated previously with tamoxifen. There were no complete tumor responses. Four patients (19%; 95% confidence interval [95% CI], 2.2%, 36%) had partial tumor responses lasting 6.3, 17.5, 23.9, and 28.1 months, respectively. Prolonged stable disease (i.e., tumor size stable for > or = 6 months) was observed in 3 patients (14%; 95% CI, 0.0%, 29%) and lasted 7.9, 12.2, and 25.1 months, respectively. Combining partial responses and prolonged stable disease yielded an overall clinical benefit rate of 33% (95% CI, 13%, 53%). Adverse events generally were consistent with the disease state; there were no serious adverse events or laboratory changes believed to be therapy-related. CONCLUSIONS: Raloxifene HCl, 150 mg, administered twice daily was safe, well tolerated, and modestly effective in highly selected postmenopausal women with advanced breast carcinoma. Further study of high dose raloxifene as monotherapy for advanced breast carcinoma most likely is unwarranted. PMID- 10813717 TI - Combined modality treatment of locally advanced breast carcinoma in elderly patients or patients with severe comorbid conditions using tamoxifen as the primary therapy. AB - BACKGROUND: The purpose of the current study was to evaluate the objective response rate and possibility of breast-conserving surgery using neoadjuvant tamoxifen in the multimodality treatment, including surgery and radiotherapy, of elderly or frail patients with locally advanced breast carcinoma. METHODS: Forty seven patients age > 75 years or age < 75 years with comorbid conditions and locally advanced breast carcinoma were treated with neoadjuvant tamoxifen (20 mg/day) for 3-6 months. This was followed by surgery and radiotherapy when feasible and adjuvant tamoxifen for 5 years or until disease recurrence. RESULTS: The median age of the patients was 72 years (range, 48-86 years). Approximately 22% had T3 lesions, 57% had T4 lesions, 22% were Stage II (AJCC Manual for Staging Cancer, 3rd edition), and 78% were Stage III. Eighty percent were estrogen receptor positive. After 6 months of treatment with neoadjuvant tamoxifen, a response rate of 47% was observed, including a complete response rate of 6%. Twenty-nine patients (62%) were rendered free of disease by surgery, including 5 with breast-conserving procedures. After a median follow-up of 40 months, 23 patients (49%) remained disease free. The median survival time had not been reached at the time of last follow-up. No major toxicity was observed, with the exception of one patient who developed a possible tamoxifen-related Stage I endometrial carcinoma. The estimated 2-year and 5-year progression free and overall survival rates were 50% and 41%, and 83% and 59%, respectively. CONCLUSIONS: The results of the current study show that neoadjuvant tamoxifen was effective in the treatment of elderly or frail patients with locally advanced breast carcinoma with estrogen receptor positive tumors, and resulted in a reasonable response rate, including complete responses and good overall survival. PMID- 10813718 TI - Expression of interleukin-6, interleukin-6 receptor, and glycoprotein 130 correlates with good prognoses for patients with breast carcinoma. AB - BACKGROUND: Interleukin-6 (IL-6) is secreted by normal epithelial breast cells but not by oncogene-transformed cells. Interleukin-6 is able to inhibit growth of breast carcinoma cells in culture. Interleukin-6 exerts its activity via two receptor subunits, IL-6R and glycoprotein 130 (gp130). The expression of these receptor subunits in breast tumors has been studied, but there are no previous reports of their prognostic significance, to the authors' knowledge. METHODS: mRNA of IL-6, IL-6R, and gp130 was studied in 75 tumor samples obtained from breast carcinoma patients. Patients were followed for a maximum of 71 months (median follow-up, 61 months; 60 patients were followed for a minimum of 5 years or died during the observation period). Prognostic factors were analyzed in univariate and multivariate analysis. RESULTS: mRNA specific to IL-6, IL-6R, and gp130 was detected in 57%, 53%, and 71% of breast carcinoma tissues, respectively. Expression was strongly correlated with earlier stages of the disease. In univariate analysis, expression of IL-6 and its receptor subunits proved to be a positive prognostic factor for overall survival (OS) and disease free survival (DFS). IL-6R and gp130 expression were good independent prognostic factors for OS. The 5-year OS of all patients was 66%. The 5-year OS in IL-6, IL 6R, and gp130 positive groups was 95%, 94%, and 90%, respectively, whereas in negative groups it was 26%, 31%, and 9%, respectively. CONCLUSIONS: Expression of IL-6, IL-6R, and gp130 in breast carcinoma tissue is associated with earlier stages of the disease. In advanced stages, expression of IL-6 and its receptor subunits predicts better prognosis. PMID- 10813719 TI - Genetic abnormalities in mammary ductal intraepithelial neoplasia-flat type ("clinging ductal carcinoma in situ"): a simulator of normal mammary epithelium. AB - BACKGROUND: Mammary ductal intraepithelial neoplasia (DIN)-flat type ("clinging ductal carcinoma in situ [DCIS]") generally is a subtle epithelial alteration characterized by one or a few layer(s) of atypical cells replacing the native epithelium. The "low power" appearance of DIN-flat type can be misinterpreted easily as "normal" because of the frequent absence of multilayered proliferation and often subtle cytologic atypia. Because it presents as an often unrecognized lesion or in association with tubular carcinoma, to the authors' knowledge the clinical and biologic significance of this lesion has not been well established. METHODS: Using polymerase chain reaction, the authors examined DNA extracts from microdissected areas of 22 cases with extensive "clinging DCIS," including 13 cases associated with infiltrating ductal carcinoma as well as 5 cases associated with more conventional types of DCIS. Eight polymorphic DNA markers with a high rate of loss of heterozygosity (LOH) in classic types of DCIS were selected to identify possible genetic alterations on chromosomes 2p, 3p, 11q, 16q, and 17q. Two cases also were used for the assessment of clonality by means of X chromosome inactivation (methylation pattern of the human androgen receptor [HUMARA] gene). RESULTS: LOH was detected in 17 of 22 lesions (77%), and monoclonality was established in the 2 cases analyzed. The most common genetic alterations were at chromosomes 11q21-23.2, 16q23.1-24.2, and 3p14.2 with LOH in 50%, 45%, and 41%, respectively, of informative cases. The DIN-flat type showed the same genetic alterations (LOH) identified in adjacent in situ and infiltrating ductal carcinoma. In contrast to the DIN-flat type, the perfectly normal mammary epithelium was associated very infrequently (1 of 16 cases; 6%) with LOH. CONCLUSIONS: The DIN-flat type represents one of the earliest, morphologically recognizable, neoplastic alterations of the breast. Recognition of the DIN-flat type is important not only for the early detection of intraductal neoplasia but also to prevent misinterpretation and utilization of this lesion as a normal control in studies. This distinctive lesion could be crucial as an explanation for at least part of the > 20% reported incidence rate of breast carcinoma recurrence observed despite ostensibly "negative" margins of breast biopsies. PMID- 10813720 TI - Combined analysis of germline polymorphisms of p53, GSTM1, GSTT1, CYP1A1, and CYP2E1: relation to the incidence rate of cervical carcinoma. AB - BACKGROUND: The authors established the genotype frequencies of cytochrome P450 (CYP1A1/MspI, CYP2E1/PstI, and CYP2E1/DraI), glutathione-S-transferase (GSTM1 and GSTT1), and p53 (exon 4/AcclI and intron 3/16-base pair duplication) gene polymorphisms in cervical carcinoma patients and controls and evaluated the association between the specific genotype or genotype combinations of these polymorphisms and the risk of cervical carcinoma. METHODS: In this case-control study, the genotypes of 181 human papillomavirus (HPV)-16 or HPV-18 positive cervical carcinoma patients and 1-to-1 age-matched controls were determined using a polymerase chain reaction-based technique. RESULTS: Among these polymorphisms, the individuals carrying arginine/proline genotypes of p53 showed a 9.5-fold increase of cervical carcinoma risk (95% confidence interval [CI], 4.9-18.6) compared with those individuals carrying arginine/arginine genotypes. The frequency of overall GSTT1 null genotypes also was significantly higher in cervical carcinoma patients compared with that of GSTT1 positive genotypes (P = 0.003; odds ratio [OR] = 1.9; 95% CI, 1.2-2.9). The genotype combination of p53 and GST played a more important role in describing the relative risk of cervical carcinoma. The individuals carrying both the arginine/proline genotype of p53 and the null genotype of GSTT1 showed a 3.5-fold increase of cervical carcinoma risk (95% CI, 1.8-7.1) compared with those individuals carrying both the arginine/arginine genotype of p53 and the GSTT1 positive genotype. In the patients who were stratified into the two age groups, the null genotypes of GSTT1 (69.1% vs. 45.5%; P = 0.016) and GSTM1 (61.8% vs. 40.0%; P = 0.028) in cervical carcinoma were significantly overrepresented in the younger age subgroup (age 40 years or younger) compared with those of controls. Especially in this age group, the individuals carrying both null genotypes of GSTT1 and GSTM1 showed a 17.8 fold increase of cervical carcinoma risk (95% CI, 2.2-141.0) compared with the individuals carrying both positive genotypes of GSTT1 and GSTM1. CONCLUSIONS: The results of the current study suggested that the arginine/proline genotype of p53, independently or in conjunction with the GSTT1 null genotype, could affect the genetic susceptibility for cervical carcinoma, and HPV positive women carrying both null genotypes of GSTT1 and GSTM1 have an increased risk of cervical carcinoma developing before age 40 years. PMID- 10813721 TI - Thiol-mediated apoptosis in prostate carcinoma cells. AB - BACKGROUND: Glutathione (GSH) maintains an optimum cellular redox potential. Chemical depletion, physical efflux from the cell, or intracellular redistribution of this thiol antioxidant is associated with the onset of apoptosis. The aim of this study was to determine the effects of a thiol depleting agent, diethylmaleate (DEM), on androgen sensitive and insensitive prostate carcinoma cells. METHODS: LNCaP and PC-3 cell lines were induced to undergo apoptosis by DEM and diamide. Apoptosis was quantified by annexin V binding and propidium iodide incorporation using flow cytometry and was confirmed by DNA gel electrophoresis. Intracellular GSH was quantified using a thiol quantitation kit and the generation of reactive oxygen intermediates was measured using dihydrorhodamine 123. Western blot assessed caspase-3, caspase-8, Bcl-2, and Bcl-XL protein expression. Mitochondrial permeability was measured using DiOC6 and stabilized using bongkrekic acid. RESULTS: DEM and diamide induced apoptosis in both androgen sensitive and insensitive cells. Apoptosis was also induced in an LNCaP transfectant cell line overexpressing Bcl-2. Apoptosis was caspase-3 dependent and caspase-8 independent. Bongkrekic acid partially prevented the effects of DEM on mitochondrial permeability but was unable to prevent the induction of apoptosis. Decreased Bcl-2 and Bci-XL protein expression was observed at the time of initial caspase-3 activation. CONCLUSIONS: This study demonstrates that thiol depletion can be used as an effective means of activating caspase-3 in both androgen sensitive and insensitive prostate carcinoma cells. Direct activation of this effector caspase may serve as a useful strategy for inducing apoptosis in prostate carcinoma cells. PMID- 10813722 TI - The use of artificial intelligence technology to predict lymph node spread in men with clinically localized prostate carcinoma. AB - BACKGROUND: The current study assesses artificial intelligence methods to identify prostate carcinoma patients at low risk for lymph node spread. If patients can be assigned accurately to a low risk group, unnecessary lymph node dissections can be avoided, thereby reducing morbidity and costs. METHODS: A rule derivation technology for simple decision-tree analysis was trained and validated using patient data from a large database (4,133 patients) to derive low risk cutoff values for Gleason sum and prostate specific antigen (PSA) level. An empiric analysis was used to derive a low risk cutoff value for clinical TNM stage. These cutoff values then were applied to 2 additional, smaller databases (227 and 330 patients, respectively) from separate institutions. RESULTS: The decision-tree protocol derived cutoff values of < or = 6 for Gleason sum and < or = 10.6 ng/mL for PSA. The empiric analysis yielded a clinical TNM stage low risk cutoff value of < or = T2a. When these cutoff values were applied to the larger database, 44% of patients were classified as being at low risk for lymph node metastases (0.8% false-negative rate). When the same cutoff values were applied to the smaller databases, between 11 and 43% of patients were classified as low risk with a false-negative rate of between 0.0 and 0.7%. CONCLUSIONS: The results of the current study indicate that a population of prostate carcinoma patients at low risk for lymph node metastases can be identified accurately using a simple decision algorithm that considers preoperative PSA, Gleason sum, and clinical TNM stage. The risk of lymph node metastases in these patients is < or = 1%; therefore, pelvic lymph node dissection may be avoided safely. The implications of these findings in surgical and nonsurgical treatment are significant. PMID- 10813723 TI - Prostate specific antigen outcome based on the extent of extracapsular extension and margin status in patients with seminal vesicle negative prostate carcinoma of Gleason score < or = 7. AB - BACKGROUND: Early (< or = 2 years) prostate specific antigen (PSA) failure after radical prostatectomy (RP) has been shown to predict for distant failure. After excluding patients with the pathologic predictors of early PSA failure, an analysis of PSA failure free (bNED) survival was performed to identify patients who may benefit from the use of postprostatectomy radiation therapy (RT). METHODS: Of 1,028 patients treated with RP for clinically localized prostate carcinoma between 1989 and 1999, 862 (84%) had either organ confined (OC), specimen confined (SC), or margin positive disease with negative seminal vesicles (SV) and a prostatectomy Gleason score < or = 7. A Cox regression multivariate analysis was performed in these patients evaluating the ability of the extent of extracapsular extension (ECE) (into but not through the capsule, SC focal ECE, SC established ECE, margin positive) and prostatectomy Gleason score (2-6 vs. 7) to predict time to postoperative PSA failure. RESULTS: SC focal ECE (P = 0.0017), SC established ECE (P < 0.0001), and margin positive disease (P < 0.0001) were significant predictors of time to postoperative PSA failure, whereas prostatectomy Gleason score and disease extending into but not through the capsule were not. Five-year bNED rates were 90%, 88%, 69%, 45%, and 33% for patients with OC, into but not through capsule, SC focal ECE, SC established ECE, and margin positive prostate carcinoma, respectively. CONCLUSIONS: Patients with SC ECE or margin positive prostate carcinoma and a prostatectomy Gleason score < or = 7 with no evidence of SV invasion may benefit from adjuvant postoperative RT. PMID- 10813724 TI - Long term survivors of childhood brain cancer have an increased risk for cardiovascular disease. AB - BACKGROUND: Cranial irradiation for children with brain tumors frequently leads to neuroendocrine deficiencies. In this controlled study, the authors investigated risk factors for cardiovascular disease (CVD) for long term survivors of childhood brain cancer. They also tested whether the presence of these risk factors was related to endocrine status. METHODS: In 26 survivors of childhood brain cancer (mean age, 25.8 years; mean posttreatment interval, 16 years) and 29 healthy controls (mean age, 27.7 years), the blood pressure, smoking habits, body mass index (BMI), and waist/hip (W/H) ratio were determined. Lipids and lipoproteins were measured and endocrine function was assessed. Carotid intima-media thickness (IMT) measurements were performed by high resolution ultrasonography. RESULTS: In the survivors of childhood brain cancer, systolic blood pressure and W/H ratio were elevated compared with controls. The cholesterol/high density lipoprotein ratio (4.7 +/- 1.7 vs. 3.4 +/- 0.8 mmol/L, P = 0.0005), low density lipoprotein cholesterol level (3.3 +/- 0.9 vs. 2.8 +/- 0.6 mmol/L, P = 0.027), and apolipoprotein B level (P = 0.001) were higher in survivors of childhood brain cancer, whereas HDL cholesterol was lower (P = 0.005). The IMT was increased in the survivor group, but only in the carotid bulb (0.63 mm +/- 1.6 vs. 0.53 mm +/- 1.1, P = 0.02), not in the internal or common carotid artery. In the absolute growth hormone deficient (GHD) population (n = 9), LDL cholesterol and apolipoprotein B levels were elevated and the W/H ratio was particularly increased compared with the other survivors of childhood brain cancer. CONCLUSIONS: For long term survivors of brain cancer, the risk for CVD is strongly increased due to dyslipidemia, central obesity, and elevated systolic blood pressure, particularly for those with GHD. The first effects of this increased risk for CVD were observed in the carotic bulb, as assessed by IMT measurements. Efforts should be directed at CVD prevention by risk factor control. PMID- 10813725 TI - Prognostic factors in chordoma of the sacrum and mobile spine: a study of 39 patients. AB - BACKGROUND: The prognosis of patients with chordoma of the sacrum and mobile spine has been reported to be dismal and attributable in the majority of cases to intralesional surgery. The purpose of this study was to evaluate the clinical outcome of these patients using modern surgical principles aimed at complete resection and to identify prognostic factors. METHODS: The clinical and morphologic features, type of surgery, and follow-up of 39 consecutive patients with chordoma were reviewed and analyzed statistically. RESULTS: Thirty sacral and 9 mobile spine chordomas (size range, 3-20 cm; mean, 8 cm) occurring in 22 women and 17 men (median age, 55 years) were analyzed. The preoperative morphologic diagnosis was based on fine-needle aspiration (FNA) biopsy, core needle biopsy, or incisional biopsy. The final surgical margins were wide in 23 patients and marginal or intralesional in 16. The mean follow-up was 8.1 years (range, 0.1-23 years). Seventeen patients (44%) developed local recurrences and 11 patients (28%) developed metastases. The estimated 5-, 10-, 15-, and 20-year survival rates were 84%, 64%, 52%, and 52%, respectively. Local recurrence was associated significantly with an increased risk of metastasis and tumor-related death (P < 0.001). CONCLUSIONS: New surgical techniques have improved local control and survival of patients with sacral or spinal chordoma significantly and have decreased progressive neurologic deterioration. Larger tumor size, performance of an invasive morphologic diagnostic procedure outside of the tumor center, inadequate surgical margins, microscopic tumor necrosis, Ki-67 > 5%, and local recurrence were found to be adverse prognostic factors. FNA is the preferred method for establishing the preoperative morphologic diagnosis of chordoma. PMID- 10813726 TI - The role of lung perfusion imaging in predicting the direction of radiation induced changes in pulmonary function tests. AB - BACKGROUND: The aim of this study was to determine whether preradiation (pre-RT) single photon emission computed tomography (SPECT) lung perfusion scans can be used to predict RT-induced changes in pulmonary function tests (PFTs). METHODS: Ninety-four patients irradiated for thoracic tumors had pre-RT SPECT lung perfusion scans. The presence of SPECT hypoperfusion distal to a central mediastinal tumor was qualitatively assessed visually without knowledge of PFT changes. Patients were grouped based on whether the diffusion capacity (DLCO) ever increased post-RT. Comparisons of patient groups were performed using 1 tailed Fisher exact tests. Patient follow-up was 6-56 months (mean, 30 months). To assess SPECT hypoperfusion objectively, the average dose to the computed tomography (CT)-defined lung was compared with the weighted-average dose (based on relative perfusion) to the SPECT-defined lung. The ratio between the CT- and SPECT-defined mean lung dose provided a quantitative assessment of hypoperfusion. The mean ratio for patients with central tumor and adjacent hypoperfusion was compared with that of the others (Wilcoxon rank-sum one-sided test). RESULTS: In patients with central tumors, 41% (9 of 22) with adjacent hypoperfusion had improvements in DLCO following radiation, versus 18% (3 of 17) of those without hypoperfusion (P = 0.11). In patients with lung carcinoma, the corresponding ratios were 40% (8 of 20) and 10% (1 of 10), respectively (P = 0.10). The mean ratio of CT dose to SPECT dose was 1.35 for patients with central tumors and adjacent hypoperfusion versus 1.16 for others (P = 0.017). CONCLUSIONS: The presence of SPECT hypoperfusion adjacent to a central mediastinal mass may identify patients likely to have improved PFTs following RT. Thus, SPECT imaging may be useful in models for predicting radiation-induced changes in PFTs. PMID- 10813727 TI - Hybrid chemotherapy consisting of cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, and vinblastine (C-MOPP/ABV) as first-line treatment for patients with advanced Hodgkin disease. AB - BACKGROUND: Combination chemotherapy, including hybrid regimens, is the standard treatment for patients with advanced Hodgkin disease (HD). Although a prolonged complete response (CR) is achieved in up to 70-80% of patients, long term complications, such as secondary leukemia, are of concern. Cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, and vinblastine (C MOPP/ABV) is a hybrid chemotherapy in which cyclophosphamide is substituted for mechlorethamine, an agent that has been implicated as the cause of secondary malignancies. METHODS: Seventy-three patients (37 males and 36 females; median age, 35 years) diagnosed with Stage III or IV HD or Stage II with bulky disease, B-symptoms, elevated erythrocyte sedimentation rate, or hilar adenopathy were treated with 8 courses of C-MOPP/ABV at a single institution during a 6-year period. Radiotherapy (RT) was administered when bulky disease or residual masses were present. Endpoints of the study were response to therapy, failure free survival (FFS), overall survival (OS), and toxicity. RESULTS: Sixty-five patients (90%) received the 8 planned courses, with 49 of them (70%) receiving the full prescribed doses. After chemotherapy, 57 patients (78%) reached CR. Seven additional patients who achieved partial response (PR) reached CR after complementary radiotherapy, with an overall CR rate of 88%. The median follow-up was 31 months. Twelve patients relapsed; the 4-year FFS was 66% (95% CI, 54-78%). Two patients died during treatment because of sepsis and four due to disease progression. The 4-year OS was 92% (95% CI, 86-98%). Age > 60 years and bone marrow involvement were related to severe infectious complications. No late toxicity was reported. CONCLUSIONS: C-MOPP/ABV induces CR with acceptable toxicity in a high proportion of advanced HD patients. PMID- 10813728 TI - Impairment of heart rate variability during paclitaxel therapy. AB - BACKGROUND: Paclitaxel, which has been reported to be effective in treating metastatic breast carcinoma and advanced ovarian carcinoma, has been associated with cardiac side effects. Therefore, the effect of paclitaxel on cardiovascular autonomic regulation was studied. METHODS: Twenty-four-hour ambulatory electrocardiogram measurements were recorded twice from 14 women with breast or ovarian carcinoma: once before paclitaxel treatment and once on the day after the second chemotherapy course. Heart rate variability (HRV) was assessed with spectral analysis. For the frequency domain analysis, HRV was assessed in the very low (0.005-0.040 hertz [Hz]), low (0.040-0.150 Hz), and high frequency (0.150-0.400 Hz) spectral components. RESULTS: The ratio between low frequency and high frequency HRV decreased (daytime values of 2.7% [standard deviation (SD) 1.6] vs. 1.7% [SD 0.91; P = 0.0098) after 2 courses of paclitaxel. The circadian fluctuation of HRV also decreased in all studied frequency components. CONCLUSIONS: The observed changes in spectral characteristics suggest that autonomic modulation of the heart rate is impaired after paclitaxel therapy. However, from these data it is not clear whether the observed changes are permanent or whether autonomic cardiac function returns to normal some time after treatment. Further studies are needed to examine whether these indices based on HRV can be used to detect those patients at risk for cardiac side effects during chemotherapy. PMID- 10813729 TI - Synucleins are expressed in the majority of breast and ovarian carcinomas and in preneoplastic lesions of the ovary. AB - BACKGROUND: The synucleins (alpha, beta, and gamma) are a family of small cytoplasmic proteins that are expressed predominantly in neurons. alpha synuclein has attracted considerable attention due to its involvement in neurodegenerative diseases. Abnormal expression of gamma synuclein has recently been reported in some breast tumors. In this study, the authors examined a panel of breast and ovarian carcinomas for expression of alpha, beta, and gamma synucleins. METHODS: Normal breast and ovary tissue samples, tissue from ovaries of women at high risk of ovarian carcinoma, and tissue from breast and ovarian carcinomas were screened by immunohistochemistry and Western blot analysis for synuclein expression. RESULTS: Synucleins were not detectable by Western blot analysis in normal breast tissue. Eighty-two percent (14 of 17) of Stage III/IV breast ductal carcinomas expressed beta synuclein, gamma synuclein, or both simultaneously. Expression of alpha synuclein was not detected in breast carcinomas by Western blot analysis. Synuclein (alpha, beta, and gamma) expression was not detectable by immunohistochemistry in normal ovarian epithelium. Eighty-seven percent (39 of 45) of ovarian carcinomas were found to express at least 1 type of synuclein, and 42% (19 of 45) expressed all 3 synucleins (alpha, beta, and gamma) simultaneously. Highly punctate gamma synuclein expression was also observed in 20% of preneoplastic lesions of the ovary, including epithelial inclusion cysts, hyperplastic epithelium, and papillary structures, suggesting that gamma synuclein up-regulation may occur early in the development of some ovarian tumors. CONCLUSIONS: alpha, beta, and gamma synuclein are expressed in a high percentage of ovarian and breast carcinomas, and abnormal gamma synuclein expression may occur early in the development of ovarian carcinoma. PMID- 10813730 TI - Validation of the Edmonton Symptom Assessment Scale. AB - BACKGROUND: The Edmonton Symptom Assessment Scale (ESAS) is a nine-item patient rated symptom visual analogue scale developed for use in assessing the symptoms of patients receiving palliative care. The purpose of this study was to validate the ESAS in a different population of patients. METHODS: In this prospective study, 240 patients with a diagnosis of cancer completed the ESAS, the Memorial Symptom Assessment Scale (MSAS), and the Functional Assessment Cancer Therapy (FACT) survey, and also had their Karnofsky performance status (KPS) assessed. An additional 42 patients participated in a test-retest study. RESULTS: The ESAS "distress" score correlated most closely with physical symptom subscales in the FACT and the MSAS and with KPS. The ESAS individual item and summary scores showed good internal consistency and correlated appropriately with corresponding measures from the FACT and MSAS instruments. Individual items between the instruments correlated well. Pain ratings in the ESAS, MSAS, and FACT correlated best with the "worst-pain" item of the Brief Pain Inventory (BPI). Test-retest evaluation showed very good correlation at 2 days and a somewhat smaller but significant correlation at 1 week. A 30-mm visual analogue scale cutoff point did not uniformly distinguish severity of symptoms for different symptoms. CONCLUSIONS: For this population, the ESAS was a valid instrument; test-retest validity was better at 2 days than at 1 week. The ESAS "distress" score tends to reflect physical well-being. The use of a 30-mm cutoff point on visual analogue scales to identify severe symptoms may not always apply to symptoms other than pain. PMID- 10813731 TI - Bone sarcomas of the head and neck in children: the St Jude Children's Research Hospital experience. AB - BACKGROUND: Bone sarcomas of the head and neck are difficult to resect. The authors reviewed their institutional experience with these tumors to characterize patients' clinical findings and to assess the impact of surgical resection on outcome. METHODS: The records of the 28 patients with bone sarcomas originating in the head and neck treated at St. Jude Children's Research Hospital between March 1962 and January 1998 were reviewed. RESULTS: There were 10 males and 18 females (median age, 12.6 years) each with a single sarcoma: osteosarcoma (18), Ewing sarcoma (7), malignant fibrous histiocytoma (MFH) (2), and fibrosarcoma (1). Primary tumor sites included the maxilla (13), skull (10), mandible (2), and other sites (3). All but one patient with Ewing sarcoma had localized disease at the time of diagnosis. All patients underwent surgery: complete resection, 8; gross total resection, 4; incomplete resection, 14; and biopsy only, 2; 22 also received chemotherapy. Radiotherapy was given to all patients with Ewing sarcoma and to four patients with primary osteosarcoma. Twelve patients survived a median of 8.4 years after diagnosis, 14 died of disease, and 2 died of unrelated causes. Local disease progression was evident in 12 patients (9 with osteosarcoma, 2 with MFH, and 1 with Ewing sarcoma) who died of disease, 9 of whom had the initial treatment of biopsy alone or incomplete resection. Patients with osteosarcoma who had the initial treatment of incomplete resection or biopsy alone were more likely to experience local failure (P = 0.001) and had poorer survival (P = 0.014) than those who underwent complete or gross total resection. CONCLUSIONS: Bone sarcomas of the head and neck are rare among children and most often are localized at the time of diagnosis. Incomplete resection of osteosarcoma is associated with local failure and poor outcome. Although aggressive surgery is essential for the cure of osteosarcoma, its necessity in the treatment of Ewing sarcomas remains controversial. PMID- 10813732 TI - Primary chondrosarcoma of the head and neck in pediatric patients: a clinicopathologic study of 14 cases with a review of the literature. AB - BACKGROUND: Primary chondrosarcoma of the head and neck in the pediatric age group is rare. The literature contains several single cases and small series; however, to the authors' knowledge, there has been no previous comprehensive larger study to evaluate the clinicopathologic aspects of these tumors. METHODS: Fourteen cases of chondrosarcoma of the head and neck from patients age 18 years or younger, diagnosed between 1970 and 1997, were retrieved from the Otorhinolaryngic-Head & Neck Tumor Registry of the Armed Forces Institute of Pathology. No secondary sarcomas (radiation-induced or arising in association with Maffucci syndrome or Ollier disease) were included. Clinical, radiographic, and histologic features were reviewed and patient follow-up obtained. RESULTS: The patients included 6 girls and 8 boys ages 3-18 years (mean, 11.8 years). Patient symptoms (nasal stuffiness or discharge, sinusitis, headaches, or a mass lesion) were related to tumor location and were present for an average of 7.2 months. No genetic abnormalities were documented. The tumors most frequently involved the maxillary sinus (n=4), followed by the mandible (n=3), nasal cavity (n=2), and neck (n=2), with 1 each of the nasopharynx, orbit, and base of the skull. The tumors ranged in size from 2.0 to 15.0 cm (mean, 3.1 cm). All tumors were invasive and malignant as determined by radiology and/or histology. The tumors were Grade 1 (n=9), Grade 2 (n=1), or Grade 3 (mesenchymal, n=2; dedifferentiated n=2). All patients were treated by surgery, followed by radiation (n=5) and/or chemotherapy (n=2). Follow-up was available for 11 patients; all were alive (at a mean of 14.8 years), with only a single patient demonstrating evidence of residual/ recurrent tumor (at 16.6 years). CONCLUSIONS: Primary head and neck chondrosarcoma in the pediatric population is typically low grade and occurs in the maxillary sinus or mandible. Despite the invasive and high grade nature of some of these tumors, there is an excellent long term prognosis for patients in this age group with tumors in these locations. PMID- 10813733 TI - Outcome for children with supratentorial primitive neuroectodermal tumors treated with surgery, radiation, and chemotherapy. AB - BACKGROUND: The outcome of a child with a primitive neuroectodermal tumors arising supratentorially (SPNET) is not well characterized and may differ from the outcome of a patient with a histologically similar cerebellar tumor (medulloblastoma [MB]). Recently, 5-year progression free survival rates as high as 80% have been reported for children with MB treated with craniospinal radiation (CRT) and chemotherapy including cisplatin, lomustine (CCNU), and vincristine (VCR). METHODS: The authors reviewed the outcome of 22 consecutive patients age 3 years and older (mean age, 10 years; range, 3-18 years) with SPNET who were treated at the study institutions between 1981 and 1996. Tumor location included was 13 pineal, 6 cortical, and 3 thalamic or suprasellar. Five patients had disease dissemination at diagnosis. All patients underwent surgery and staging, followed by CRT and chemotherapy with cisplatin, CCNU, and VCR. RESULTS: Of the 22 patients, 13 had developed disease progression and 10 had died at the time of last follow-up. Overall progression free survival (PFS) was 47% +/- 11% at 3 years and 37% +/- 11% at 5 years. There was a significant difference in PFS between patients with localized disease versus those with disseminated disease (P = 0.04). There was no statistical association between tumor location and survival. Although not significant (P = 0.21), there was a trend toward better survival of those patients with complete or near-complete resection compared with those with partial resection or biopsy. CONCLUSIONS: The results of the current study demonstrate that the outcome for children with SPNET treated with radiation and chemotherapy appears worse than for children with MB treated with identical therapy. This suggests that there may be biologic differences between supratentorial and infratentorial primitive neuroectodermal tumors, thus requiring refinements in treatment. PMID- 10813735 TI - Author reply. PMID- 10813734 TI - Failure to confirm major objective antitumor activity for streptozocin and doxorubicin in the treatment of patients with advanced islet cell carcinoma. PMID- 10813736 TI - Intratesticular leiomyosarcoma in a young man after high dose doping with oral turinabol. A case report. PMID- 10813737 TI - Author reply. PMID- 10813738 TI - High molecular weight kininogen inhibition of endothelial cell function on biomaterials. AB - Synthetic vascular grafts implanted into humans fail to develop a complete endothelial lining. In previous studies, we have shown that high-molecular-weight kininogens (HMWK) adsorb to the surfaces of biomaterials. In addition, it has been demonstrated that these proteins modulate cellular function. In the present study, we report on the adhesion and proliferation of human umbilical-vein endothelial cells (HUVEC) on tissue culture polystyrene, glass, polyurethane, and Mylar(trade mark) surfaces coated with human HMWK, either single-chain HMWK (SC HMWK) or double-chain HMWK (DC-HMWK). Surfaces coated with fibronectin served as a positive control for these experiments. Parallel experiments were performed in which HUVEC were allowed to migrate from crosslinked dextran microcarrier beads (Cytodex 2) onto HMWK-coated surfaces. Our results indicate that HMWK-coated surfaces inhibit endothelial cell adhesion, proliferation, and migration at 24 and 72 h, and this inhibition is concentration dependent. To determine a potential mechanism for this inhibitory phenomenon, cells were stained for cytoskeletal actin filaments using rhodamine-phalloidin. Endothelial cells on HMWK-coated surfaces displayed F-actin filament reorganization/disassembly, characterized by the absence of peripheral actin bands in focal adhesion contacts. We conclude that HMWK inhibit endothelial cell adhesion, proliferation, and migration on a variety of biomaterial surfaces. This inhibitory effect may play a role in promoting the lack of endothelialization in synthetic vascular grafts, which is thought to play a significant role in the failure of these devices. PMID- 10813739 TI - Targeted drug delivery to C6 glioma by transferrin-coupled liposomes. AB - Recent advances in liposome technology have shown promise relative to the introduction of chemotherapeutic agents with reduced toxicity, extended longevity, and potential for cell-specific targeting. In this study we report the engineering of a liposomal delivery system for the chemotherapeutic drug doxorubicin. The system was targeted specifically to C6 glioma in vitro by coupling transferrin to the distal ends of liposomal polyethylene glycol (PEG) chains. The transferrin receptor is overexpressed on glioma, with the extent of overexpression correlated to the severity of the tumor. Significantly increased gliomal doxorubicin uptake was achieved by drug encapsulation within transferrin coupled liposomes compared to other liposome populations. Doxorubicin encapsulated within transferrin-coupled liposomes exhibited 70% of free doxorubicin uptake as compared to 54, 14, and 34% for non-PEG, PEG, and albumin coupled PEG liposomes, respectively. Competitive binding assays support the receptor-mediated mechanism of targeting. The addition of one microM free transferrin reduced the uptake of doxorubicin encapsulated within transferrin coupled liposomes by 30%. PMID- 10813740 TI - Morphometric and mechanical evaluation of titanium implant integration: comparison of five surface structures. AB - Achieving a stable bone-implant interface is an important factor in the long-term outcome of joint arthroplasty. In this study, we employed an ovine bicortical model to compare the bone-healing response to five different surfaces on titanium alloy implants: grit blasted (GB), grit blasted plus hydroxyapatite (50 microm thick) coating (GBHA), Porocoat(R) (PC), Porocoat(R) with HA (PCHA) and smooth (S). Push-out testing, histology, and backscatter scanning electron microscope (SEM) imaging were employed to assess the healing response at 4, 8, and 12 weeks. Push-out testing revealed PC and PCHA surfaces resulted in significantly greater mechanical fixation over all other implant types at all time points (p <.05). HA coating on the grit-blasted surface significantly improved fixation at 8 and 12 weeks (p <.05). The addition of HA onto the porous coating did not significantly improve fixation in this model. Quantification of ingrowth/ongrowth from SEM images revealed that HA coating of the grit-blasted surfaces resulted in significantly more ongrowth at 4 weeks (p <.05). PMID- 10813741 TI - Evolution of porosity during in vitro hydroxycarbonate apatite growth in sol-gel glasses. AB - A bioactive glass of composition (mol %) SiO(2) 58, CaO 36, P(2)O(5) 6 was obtained using the sol-gel method. Changes in porosity during the growth of a hydroxycarbonate apatite (HCA) phase were studied at different stages of the process. The high Ca (II) content led to a higher degree of porosity and enhancement of Ca(2+) released from surface and bulk when the glass was soaked into simulated body fluid (SBF). Saturation of the media (SBF) together with the porosity of the material led to fast growth of HCA on the surface and into the pores. PMID- 10813742 TI - Optimization of ligand presentation for immunoadsorption using star-configured polyethylene glycols. AB - Medical, pharmaceutical, and industrial applications of immunoadsorption are frequently limited by the technologic problems of low affinity, inadequate capacity, hydrophobicity, and bioincompatibility. To overcome these difficulties we studied the use of star-burst configured polyethylene glycols (star-PEGs) with immunoreactive molecules covalently bound to the end of each of the multiple flexible arms. The optimum pH ranges were determined to maintain stability of the tresyl chloride modified star reagents, and the chemistry was designed for their subsequent linkage to the immunoadsorbent moiety. We then devised the chemical reactions using nitration or hydrazine activation to tether these 64-arm structures to polymer supports made of polysulfone or polymethylmethacrylate, respectively. Transmission, scanning, and atomic force microscopy confirmed the preservation of the star configuration, even after linkage to the luminal surface of hollow fiber devices. To establish that these modified devices also maintained immunoadsorption reactivity, we used a model having relevance for human autoimmune disease and demonstrated the clearance of antihistone antibodies by tethered histones. This novel approach to increasing the capacity of immunoadsorption benefits from the star configuration which provides a high density of ligand, improved hydrophilicity of the surface, spacing of reactive molecules away from the support structure, and possible optimization of epitope immunoreactivity by arm-to-arm interaction of the bound molecules. PMID- 10813743 TI - Quantitative comparison of bone growth behavior in granules of Bioglass, A-W glass-ceramic, and hydroxyapatite. AB - The hypothesis that bioactive glass particulate increases the rate of bone proliferation over that of synthetic hydroxyapatite and bioactive glass-ceramic was tested in these experiments. Three types of bioactive particles-45S5 Bioglass(R), synthetic hydroxyapatite, and A-W glass-ceramic-were implanted in 6 mm-diameter holes drilled in the femoral condyles of mature rabbits. Bone growth rate was measured using an image processor. 45S5 Bioglass(R) produced bone more rapidly than either A-W glass-ceramic or hydroxyapatite. At the later time periods, 45S5 Bioglass(R) was resorbed more quickly than A-W glass-ceramic. Synthetic hydroxyapatite was not resorbed at all. Backscattered electron imaging suggested that the resorption process occurred by solution-mediated dissolution, which produced chemical changes in the enclosed particulate. It was concluded that the rate of bone growth correlates with the rate of dissolution of silica as the particles resorb. PMID- 10813744 TI - Evaluation by electrochemical tests of the passive film stability of equiatomic Ni-Ti alloy also in presence of stress-induced martensite. AB - In this study, potentiodynamic polarization scans, potentiostatic scratch tests, and modified American Society for Testing and Materials F746 tests were carried out in simulated body fluids on commercial orthodontic wires made of different classes of materials and on titanium used as a reference. The stability of passivating film, evaluated by electrochemical techniques that abruptly damage it, e.g., potentiostatic scratch test, increased in the following order: Ni-Ti NaSCN >Na(2)SO(4) >KBr >KCl >KF. A comparison of the effects of various potassium salts revealed that anions were chiefly responsible in stabilizing HSA. The above series was found similar to the electroselectivity series of anions towards the anion-exchange resins and reverse of the Hofmeister series, suggesting that preferential binding of anions to HSA rather than hydration, was primarily responsible for stabilization. Further, single-step transition observed with GdnHCl can be ascribed to its ionic character as the free energy change associated with urea denaturation in the presence of 1.0 M KCl (5,980 cal/mol) was similar to that obtained with GdnHCl (5,870 cal/mol). PMID- 10813829 TI - Steady-state and time resolved fluorescence of albumins interacting with N oleylethanolamine, a component of the endogenous N-acylethanolamines. AB - The functions of N-acylethanolamines, minor constituents of mammalian cells, are poorly understood. It was suggested that NAEs might have some pharmacological actions and might serve as a cytoprotective response, whether mediated by physical interactions with membranes or enzymes or mediated by activation of cannabinoid receptors. Albumins are identified as the major transport proteins in blood plasma for many compounds including fatty acids, hormones, bilirubin, ions, and many drugs. Moreover, albumin has been used as a model protein in many areas, because of its multifunctional binding properties. Bovine (BSA) and human (HSA) serum albumin are similar in sequence and conformation, but differ for the number of tryptophan residues. This difference can be used to monitor unlike protein domains. Our data suggest that NOEA binds with high affinity to both albumins, modifying their conformational features. In both proteins, NOEA molecules are linked with higher affinity to hydrophobic sites near Trp-214 in HSA or Trp-212 in BSA. Moreover, fluorescence data support the hypothesis of the presence of other NOEA binding sites on BSA, likely affecting Trp-134 environment. The presence of similar binding sites is not measurable on HSA, because it lacks of the second Trp residue. PMID- 10813830 TI - Role of the minor energetic determinants of chicken egg white lysozyme (HEWL) to the stability of the HEWL.antibody scFv-10 complex. AB - Seven of the 13 non-glycine contact amino acids in the hen (chicken) egg white lysozyme (HEWL) epitope for antibody Fab-10 each contribute < or =0.3 kcal/mol to the change in free energy (DeltaDeltaG(D)) from wild type (WT) when replaced by alanine (nullspots), and three others each give (0.7 < DeltaDeltaG(D) < or = 1. 0) kcal/mol (warm spots) (Rajpal et al. Protein Sci 1998;7:1868-1874). The low DeltaDeltaG(D) values introduced by alanine mutations present an opportunity to explore accurately their cumulative effects, as the sum of the combined DeltaDeltaG(D) values is not so large as to destabilize the complex beyond the range of accurate measurement. Substitution of six of the seven null spot residues by alanine leads to a cumulative DeltaDeltaG(D) = 2.25 +/- 0.04 kcal/mol, whereas the sum of the six individual changes is only -0.36 +/- 0.32 kcal/mol. The triple warm spot mutation generates a DeltaDeltaG(D) = 5.11 +/- 0.06 kcal/mol versus DeltaDeltaG(D) = 2.52 +/- 0.22 kcal/mol for the sum of the three individuals. The non-additivity in the individual DeltaDeltaG(D) values for the alanine mutations may indicate that these residues provide a conformationally stabilizing effect on the hot spot residues, each of which exhibits DeltaDeltaG(D) > 4.0 kcal/mol on alanine substitution. PMID- 10813831 TI - Lower kinetic limit to protein thermal stability: a proposal regarding protein stability in vivo and its relation with misfolding diseases. AB - In vitro thermal denaturation experiments suggest that, because of the possibility of irreversible alterations, thermodynamic stability (i.e., a positive value for the unfolding Gibbs energy) does not guarantee that a protein will remain in the native state during a given timescale. Furthermore, irreversible alterations are more likely to occur in vivo than in vitro because (a) some irreversible processes (e.g., aggregation, "undesirable" interactions with other macromolecular components, and proteolysis) are expected to be fast in the "crowded" cellular environment and (b) in many cases, the relevant timescale in vivo (probably related to the half-life for protein degradation) is expected to be longer than the timescale of the usual in vitro experiments (of the order of minutes). We propose, therefore, that many proteins (in particular, thermophilic proteins and "complex" proteins systems) are designed (by evolution) to have significant kinetic stability when confronted with the destabilizing effect of irreversible alterations. We show that, as long as these alterations occur mainly from non-native states (a Lumry-Eyring scenario), the required kinetic stability may be achieved through the design of a sufficiently high activation barrier for unfolding, which we define as the Gibbs energy barrier that separates the native state from the non-native ensemble (unfolded, partially folded, and misfolded states) in the following generalized Lumry-Eyring model: Native State <--> Non-Native Ensemble --> Irreversibly Denatured Protein. Finally, using familial amyloid polyneuropathy (FAP) as an illustrative example, we discuss the relation between stability and amyloid fibril formation in terms of the above viewpoint, which leads us to the two following tentative suggestions: (a) the hot spot defined by the FAP-associated amyloidogenic mutations of transthyretin reflects the structure of the transition state for unfolding and (b) substances that decrease the in vitro rate of transthyretin unfolding could also be inhibitors of amyloid fibril formation. PMID- 10813832 TI - On the design and analysis of protein folding potentials. AB - Pairwise interaction models to recognize native folds are designed and analyzed. Different sets of parameters are considered but the focus was on 20 x 20 contact matrices. Simultaneous solution of inequalities and minimization of the variance of the energy find matrices that recognize exactly the native folds of 572 sequences and structures from the protein data bank (PDB). The set includes many homologous pairs, which present a difficult recognition problem. Significant recognition ability is recovered with a small number of parameters (e.g., the H/P model). However, full recognition requires a complete set of amino acids. In addition to structures from the PDB, a folding program (MONSSTER) was used to generate decoy structures for 75 proteins. It is impossible to recognize all the native structures of the extended set by contact potentials. We therefore searched for a new functional form. An energy function U, which is based on a sum of general pairwise interactions limited to a resolution of 1 angstrom, is considered. This set was infeasible too. We therefore conjecture that it is not possible to find a folding potential, resolved to 1 angstrom, which is a sum of pair interactions. PMID- 10813833 TI - Crystal structure of YbaK protein from Haemophilus influenzae (HI1434) at 1.8 A resolution: functional implications. AB - Structural genomics of proteins of unknown function most straightforwardly assists with assignment of biochemical activity when the new structure resembles that of proteins whose functions are known. When a new fold is revealed, the universe of known folds is enriched, and once the function is determined by other means, novel structure-function relationships are established. The previously unannotated protein HI1434 from H. influenzae provides a hybrid example of these two paradigms. It is a member of a microbial protein family, labeled in SwissProt as YbaK and ebsC. The crystal structure at 1.8 A resolution reported here reveals a fold that is only remotely related to the C-lectin fold, in particular to endostatin, and thus is not sufficiently similar to imply that YbaK proteins are saccharide binding proteins. However, a crevice that may accommodate a small ligand is evident. The putative binding site contains only one invariant residue, Lys46, which carries a functional group that could play a role in catalysis, indicating that YbaK is probably not an enzyme. Detailed sequence analysis, including a number of newly sequenced microbial organisms, highlights sequence homology to an insertion domain in prolyl-tRNA synthetases (proRS) from prokaryote, a domain whose function is unknown. A HI1434-based model of the insertion domain shows that it should also contain the putative binding site. Being part of a tRNA synthetases, the insertion domain is likely to be involved in oligonucleotide binding, with possible roles in recognition/discrimination or editing of prolyl-tRNA. By analogy, YbaK may also play a role in nucleotide or oligonucleotide binding, the nature of which is yet to be determined. PMID- 10813834 TI - Assignment of enzyme substrate specificity by principal component analysis of aligned protein sequences: an experimental test using DNA glycosylase homologs. AB - We have studied the relationship between amino acid sequence and substrate specificity in a DNA glycosylase family by characterizing experimentally the specificity of four new members of the family. We show that principal component analysis (PCA) of the sequence family correctly predicts the substrate specificity of one of the novel homologs even though conventional sequence analysis methods fail to group this homolog with other sequences of the same specificity. PCA also suggested, correctly, that another homolog characterized previously differs in its specificity from those sequences with which it clusters by conventional criteria. These results suggest that principal component analysis of sequence families can be a useful tool in annotating genome sequences when there is ambiguity concerning which subfamily a new homolog belongs to. Published 2000 Wiley-Liss, Inc. PMID- 10813835 TI - Zinc binding in bovine alpha-lactalbumin: sequence homology may not be a predictor of subtle functional features. AB - alpha-Lactalbumin (alpha-LA), a calcium-binding protein, also possesses zinc binding sites comprising a single strong site and several weaker secondary sites. The only site found by X-ray crystallography (Ren et. al., J. Biol. Chem. 1993;268:19292) was Glu 49 of human alpha-LA, but zinc binding had never been measured in solution for human alpha-LA. This residue was genetically substituted by Ala in bovine alpha-LA and the metal-binding properties of the resulting desMetE49A protein were compared with those for native alpha-LA by fluorescence methods. Surprisingly, desMetE49A alpha-LA and the native bovine protein had similar affinities for both Zn(2+) and Ca(2+). Genetic substitution of other possible candidates for Zn(2+) chelating residues, which included Glu 25, did not alter the affinity of bovine alpha-LA to Zn2+; however, substitution of Glu 1 by Met resulted in the disappearance of strong Zn(2+) binding. A proposed site involves Glu 1, Glu 7, Asp 11, and Asp 37, which would participate in strong Zn(2+) binding based on their propinquity to Glu 1. Human alpha-LA, which has a Lys at position 1 rather than Glu, binds zinc with a reduced affinity compared with native bovine alpha-LA, suggesting that the site identified from the X-ray structure did not correspond to strong zinc binding in solution. PMID- 10813836 TI - Aromatic interactions in homeodomains contribute to the low quantum yield of a conserved, buried tryptophan. AB - Trp 48, a conserved, buried residue commonly found in the hydrophobic core of homeodomains, has an unusually low fluorescence quantum yield. Chemical denaturation of Drosophila homeodomains Engrailed and Antennapedia(C39S) result in a four-fold increase in quantum yield, while unfolding of Ultrabithorax causes a twenty-fold enhancement. Global analysis of time-resolved fluorescence decay monitored at multiple emission wavelengths reveals sub-nanosecond lifetime components which dominate the overall intensity. Based on structure and sequence analysis of several homeodomains, we deduce that quenching is due to a transient, excited-state NH ellipsis pi hydrogen bond involving Trp 48 and a conserved aromatic residue at position 8. Additionally, both time-resolved fluorescence of indole-benzene mixtures and an electrostatic model of the proposed tryptophan aromatic interaction substantiate different aspects of this mechanism. A survey of the Protein Data Bank reveals many proteins with tryptophan-aromatic pairs where the indole nitrogen participates in a NH ellipsis pi hydrogen bond with the ring of another aromatic residue. Chemical denaturation of one protein found in this survey, human fibronectin type III module 10, causes an enhancement of the fluorescence quantum yield. This unique interaction has implications for many other systems and may be useful for studying larger, multi-tryptophan containing proteins. PMID- 10813837 TI - Characterization of the active site of group B streptococcal hyaluronan lyase. AB - Hyaluronan lyase is secreted by most strains of the human pathogen, group B streptococcus. Site-directed mutagenesis of the enzyme identified three amino acid residues important for enzyme activity, H479, Y488, and R542. These three residues are in close proximity in the putative active site of a homology model of group B streptococcal hyaluronan lyase. The homology model was based on the crystal structure of another related glycosaminoglycan lyase, chondroitin AC lyase, which exhibits different substrate specificity. Two asparagine residues in the active site groove, N429 and N660, were also found to be essential for enzyme activity. In addition, conversion of two adjacent tryptophan residues in the groove to alanines abolished activity. All amino acids found to be essential in GBS hyaluronan lyase are conserved in both enzymes. However, several amino acids in the active site groove of the two enzymes are not conserved. In the 18 cases in which one of these amino acids in GBS hyaluronan lyase was replaced with its corresponding amino acid in chondroitin AC lyase, no major loss of activity or change in substrate specificity was observed. PMID- 10813838 TI - A novel exhaustive search algorithm for predicting the conformation of polypeptide segments in proteins. AB - We present a fast ab initio method for the prediction of local conformations in proteins. The program, PETRA, selects polypeptide fragments from a computer generated database (APD) encoding all possible peptide fragments up to twelve amino acids long. Each fragment is defined by a representative set of eight straight phi/psi pairs, obtained iteratively from a trial set by calculating how fragments generated from them represent the protein databank (PDB). Ninety-six percent (96%) of length five fragments in crystal structures, with a resolution better than 1.5 A and less than 25% identity, have a conformer in the database with less than 1 A root-mean-square deviation (rmsd). In order to select segments from APD, PETRA uses a set of simple rule-based filters, thus reducing the number of potential conformations to a manageable total. This reduced set is scored and sorted using rmsd fit to the anchor regions and a knowledge-based energy function dependent on the sequence to be modelled. The best scoring fragments can then be optimized by minimization of contact potentials and rmsd fit to the core model. The quality of the prediction made by PETRA is evaluated by calculating both the differences in rmsd and backbone torsion angles between the final model and the native fragment. The average rmsd ranges from 1.4 A for three residue loops to 3.9 A for eight residue loops. PMID- 10813839 TI - Molecular dynamics simulation of hen egg white lysozyme: a test of the GROMOS96 force field against nuclear magnetic resonance data. AB - Biomolecular force fields for use in molecular dynamics (MD) simulations of proteins, DNA, or membranes are generally parametrized against ab initio quantum chemical and experimental data for small molecules. The application of a force field in a simulation of a biomolecular system, such as a protein in solution, may then serve as a test of the quality and transferability of the force field. Here, we compare various properties obtained from two MD simulations of the protein hen egg white lysozyme (HEWL) in aqueous solution using the latest version, GROMOS96, of the GROMOS force field and an earlier version, GROMOS87+, with data derived from nuclear magnetic resonance (NMR) experiments: NOE atom atom distance bounds, (3)J(HNalpha)-coupling constants, and backbone and side chain order parameters. The convergence of these quantities over a 2-ns period is considered, and converged values are compared to experimental ones. The GROMOS96 simulation shows better agreement with the NMR data and also with the X-ray crystal structure of HEWL than the GROMOS87+ simulation, which was based on an earlier version of the GROMOS force field. PMID- 10813840 TI - Factors determining the relative stability of anionic tetrahedral complexes in serine protease catalysis and inhibition. AB - Quantum mechanical ab initio (RHF/6-31+G*//RHF/3-21G) calculations were used to simulate the formation of the tetrahedral complex intermediate (TC) in serine protease active site by substrates and transition-state analog inhibitors. The enzyme active site was simulated by an assembly of the amino acids participating in catalysis, whereas the substrates and inhibitors were simulated by small ligands, acetamide (1) and trifluoroacetone (2), respectively. For the first time, the principal factors determining the relative stability of the TC in serine proteases are arranged according to their energy contributions. These include (a) formation of the new covalent bond between Ser195 O(gamma) and the electrophilic center of a ligand; (b) stabilization of the oxyanion in the oxyanion hole; (c) basic catalysis by His57; and (d) hydrogen bond between Asp102 carboxylate and N(delta) of the protonated His57. We have directly calculated the gas-phase relative free energy of formation of TC(AS)(2) and TC(AS)(1), the value of DeltaDeltaG(g)[TC(AS)(2,1)]. It is DeltaE(cov), the relative energy of the new covalent bond between the enzyme and the ligand formed in a TC that determines the experimentally observed large difference in the stability of TCs formed by substrates and TS-analog inhibitors of serine proteases. We demonstrated that the relative stability of TCs formed by a series of mono- and dipeptide amides and TFKs, derived from experimental kinetic data, can be rather well approximated by the sum of the theoretically calculated value of DeltaDeltaG(g)[TC(AS)(2, 1)] and the difference in hydration free energies of isolated ligands. PMID- 10813841 TI - Disulfide bond plasticity in epidermal growth factor. AB - Epidermal growth factor (EGF) has a (1-3,2-4,5-6) disulfide-bonding pattern. This pattern is found in nearly all EGF-like domains, despite wide variation in sequences. Biological data from EGF and at least one EGF-like domain show that disulfide bond isomers have significant bioactivity and suggests that the EGF fold can accommodate alternate disulfide-bonding patterns. The disulfide bonds in murine EGF were altered to seven different patterns and structures were calculated incorporating all the restraints from the highest resolution restraint set available (Tejero et al., 1996). Results showed that besides the native (1 3,2-4,5-6), two other disulfide-bonding patterns: (1-2,3-4,5-6) and (1-3,2-5,4-6) satisfied the restraints as well as the native. The results for these two patterns were indistinguishable from the native on the basis of distance and dihedral violations, XPLOR energies, Procheck statistics, and RMSDs of the final set of structures. Two other disulfide bond patterns, (1-2,3-5, 4-6) and (1-4,2 3,5-6) were able to satisfy all the distance restraints but had one or more cysteine dihedral violations. For all seven isomers, the final calculated structures were highly similar to EGF with all-atom RMSD's in the 1. 5-2 A range. These results suggest that the EGF backbone fold has the unique property of accommodating several different disulfide-bonding patterns. PMID- 10813842 TI - DNA microarray analyses of genes regulated during the differentiation of embryonic stem cells. AB - Embryonic stem (ES) cells are derived from the inner cell mass of blastocysts, and in response to retinoic acid (RA) are induced to differentiate to form some of the first distinguishable cell types of early mammalian development. This makes ES cells an attractive model system for studying the initial developmental decisions that occur during embryogenesis and the molecular genetics and associated mechanisms underlying these decisions. Additionally, ES cells are of significant interest to those characterizing various gene functions utilizing transgenic and gene-targeting techniques. With the advent of DNA microarray technology, which allows for the study of expression patterns of a large number of genes simultaneously within a cell type, there is an efficient means of gaining critical insights to the expression, regulation, and function of genes involved in mammalian development for which information is not currently available. To this end, we have utilized Clontech's Atlas Mouse cDNA Expression Arrays to examine the expression of 588 known regulatory genes in D3 ES cells and their RA-induced differentiated progeny. We report that nearly 50% of the regulatory genes are expressed in D3 and/or D3-differentiated cells. Of these genes, the steady-state levels of 18 are down-regulated and 61 are up-regulated by a factor of 2.5-fold or greater. These changes in gene expression are highly reproducible and represent changes in the expression of a variety of molecular markers, including: transcription factors, growth factors and their receptors, cytoskeletal and extracellular matrix proteins, cell surface antigens, and intracellular signal transduction modulators and effectors. PMID- 10813843 TI - GATA-1 is a potential repressor of anti-Mullerian hormone expression during the establishment of puberty in the mouse. AB - Anti-Mullerian hormone (AMH), also known as Mullerian inhibiting substance (MIS), is one of the earliest and best-known markers of Sertoli cell differentiation and is expressed until around puberty. The present study is aimed at the better understanding of the molecular pathways involved in testicular development and establishment of adult functions with regards to AMH regulation. We found, within the mouse AMH promoter, putative GATA motifs (A/T)GATA(A/G), known to be specifically bound by members of the GATA transcription factor family. We then carried out RNase protection assays and immunohistochemical techniques aimed at comparing precisely the chronological expression patterns of AMH and GATA-1, this latter being expressed in the testis after birth. Using both approaches we found an inverse and close relationship between AMH and GATA-1 mRNA and protein expression during the pre-pubertal period. These results allowed us to define a transitory 4-5-day period, starting from 3 dpp when both proteins are heterogeneously expressed in Sertoli cells and showed that the appearance of GATA 1 is associated with the decrease of AMH expression in these cells. Furthermore DNA-protein interaction in in vitro studies showed first that GATA-1 binds with various affinities on sites found in the AMH promoter and second that the proximity of the two strongest affinity sites leads to a synergistic binding effect. Altogether, the present study suggests that GATA-1 participates in AMH gene repression during the pre-pubertal period. PMID- 10813844 TI - Translation of a unique transcript for protein isoaspartyl methyltransferase in haploid spermatids: implications for protein storage and repair. AB - The mammalian testis contains high levels of a protein, L-isoaspartyl (D aspartyl) O-methyltransferase (PIMT), postulated to play a role in the repair of age-damaged proteins. To examine the regulation of PIMT concentrations during the development of spermatozoa, poly(A)+ RNA was isolated from purified populations of pachytene spermatocytes and round spermatids. Northern blot analysis revealed that a unique 1.1-1.3 kb PIMT transcript is present in preparations of round spermatid and pachytene spermatocyte poly (A)+ RNA. The concentration of this small PIMT transcript is at least four times higher in mRNA isolated from round spermatids than in mRNA isolated from pachytene spermatocytes, indicating that the PIMT gene is actively transcribed during the haploid phase of spermatogenesis. The germ cell-specific PIMT transcripts are distributed between the polysomal fraction and the nonpolysomal fractions of testis RNA, suggesting that translational controls also contribute to the high concentrations of PIMT in mammalian sperm. PIMT function is not essential for spermatogenesis because the testes from transgenic mice lacking PIMT activity have normal levels of protamine transcripts, and because functional sperm can be recovered from the cauda epididymis. The protein repair function of the PIMT may be more important in maintaining the fertilization competence of translationally-inactive mature sperm during the prolonged period of epididymal transit and storage in the male reproductive tract. PMID- 10813845 TI - Production of transgenic porcine blastocysts by nuclear transfer. AB - In this study the in vitro development of porcine nuclear transfer (NT) embryos was investigated. Transgenic fetal fibroblast cells that were frozen after 5 days of serum starvation were injected immediately after thawing into enucleated metaphase II (MII) oocytes. Reconstructed embryos were activated by incubation in 200 microM thimerosal followed by a 30-min treatment of 8 mM DTT. The embryos were subsequently cultured in NCSU23, supplemented with 4 mg/ml BSA for 7 days. The actual cleavage rate (embryos showing > or =2 nuclei) in 6 replicates was 33% (ranging from 15% to 50%). Three blastocysts with cell numbers of 14, 15, and 18 were obtained. The blastocyst rate was significantly lower for NT embryos as opposed to parthenogenetically activated embryos (1% vs. 5%; P<0.05). The neomycin-resistance gene was amplified by PCR in all three NT embryos, indicating their origin from the injected transgenic fibroblasts. Efforts are now being directed in improvements in the nuclear transfer technology, whereby viable fetuses or offspring can be produced from these NT-embryos. PMID- 10813846 TI - Transfer of foreign gene to giant freshwater prawn (Macrobrachium rosenbergii) by spermatophore-microinjection. AB - We developed a spermatophore-microinjection (SMI) technique that allows exogenous DNA fragments to be transferred easily into the giant freshwater prawn (Macrobrachium rosenbergii), an important aquacultural shellfish and aquatic invertebrate model. From 28 to 1, 000 ng of the circular plasmid pGL, in a total volume of 1 microl, were directly microinjected into spermatophores. Fertilization and hatching of prawns created with SMI were completed in vivo. Fertilization and hatching rates in the SMI treatments did not differ from those of the untreated control group. The genomes of free swimming, SMI-created larvae (21 days after fertilization) were analyzed using PCR and Southern blot analyses. A product with a molecular mass of 680 bp was amplified. It corresponded to amplifications of pGL, and Southern blot analysis revealed that the amplified band was positive. The gene transfer rate was primarily dependent on the concentration of DNA during SMI. The higher the concentration of pGL, the higher the rate of gene transfer. PCR and Southern blot analyses detected the existence of foreign DNA in 16 of 23 samples (70%) of genomic DNA isolated from hatched larvae in the 750 ng pGL SMI treatment. SMI, described here for the first time, is the simplest and most efficient method for mass producing transgenic giant freshwater prawns. PMID- 10813847 TI - Interaction of cell cycle kinases, microtubules, and chromatin in ascidian oocytes during meiosis. AB - We used kinase assays and confocal microscopy to study the interaction of cell cycle proteins with microtubule organising centres (MTOC) and chromatin in ascidian oocytes during meiosis. The activity of maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK) appear not to be correlated in control oocytes. MPF activity peaks during metaphase I and II of the meiotic cell cycle whereas the activity of MAPK peaks at telophase I and is subsequently degraded to remain at low levels for the remainder of meiosis. The protein synthesis inhibitor emetine induces the degradation in MPF activity in unfertilized metaphase-I (M-I) oocytes, while MAPK is unaffected. Emetine does not alter the activities of these cell cycle kinases in fertilized oocytes during meiosis I but MPF activity remains low while MAPK activity is high for an elongated time period and oocytes do not complete meiosis I. Emetine induces maternal MTOC duplication in unfertilized M-I oocytes and prevents sperm aster growth in fertilized oocytes, but it does not alter the M-I meiotic apparatus in unfertilized oocytes. These experiments suggest that neither MPF alone nor emetine-sensitive proteins are responsible for M-I arrest in ascidian oocytes, MAPK may ensure this stability. In addition, we showed that the maternal MTOC is present at M-I but suppressed from duplicating in an emetine-sensitive manner. PMID- 10813848 TI - Oocyte metabolism predicts the development of cat embryos to blastocyst in vitro. AB - Current methods for detecting complete oocyte maturation and developmental competence are inadequate. The objectives of this study were to (1) examine the relationship between cat oocyte energy metabolism and development in vitro after fertilization and (2) determine if cumulus cell metabolism could be used to predict development of individual oocytes after fertilization in vitro. The hanging drop method was used to assess metabolism of three different types of cat oocytes: immature (IMO), in vitro matured (IVM), and in vivo matured (IVOM). Stage of oocyte nuclear maturation or developmental competence was assessed after metabolic analysis. Glycolysis and oxidation of glucose, glutamine, palmitate, and lactate increased with the resumption of oocyte meiotic maturation (P<0.05). Pyruvate was the preferred substrate, but uptake was not linked to maturation. IVM oocytes had impaired glucose and palmitate metabolism compared to IVOM oocytes (P<0.05). Oocyte glycolytic activity and oocyte glucose oxidation correlated well with embryo development after insemination in vitro (P<0.05). Furthermore, oocytes that had similar glucose metabolism and that were grouped together for culture on this basis had higher (P<0.05) overall rates of development than oocytes grouped randomly. There was no correlation (P>0.05) between cumulus cell metabolism and individual oocyte development after in vitro fertilization. The data reveal that energy metabolism is linked to oocyte maturation in the cat and that glucose metabolic activity can indicate those oocytes most likely to fertilize and develop in vitro. Measuring cumulus cell metabolism does not accurately predict individual oocyte development after insemination in vitro. PMID- 10813849 TI - Adenosine blocks hormone-induced meiotic maturation by suppressing purine de novo synthesis. AB - We have examined adenosine (Ado) suppression of FSH-induced germinal vesicle breakdown (GVB) and its relationship to purine de novo synthesis. Oocyte-cumulus cell complexes (OCC) from PMSG-primed, immature mice were cultured 17-18 hr in medium containing 4 mM hypoxanthine (HX) or 300 microM dibutyryl cAMP (dbcAMP) to maintain meiotic arrest, and FSH was added to stimulate meiotic maturation. In the absence of FSH, Ado (1-250 microM) had no effect in dbcAMP-arrested oocytes but dose-dependently suppressed maturation in HX-treated oocytes. FSH-induced maturation was prevented by Ado, though more effectively in dbcAMP-supplemented cultures. Ado affected the magnitude, but not the kinetics pattern, of the response to FSH. Inosine also blocked meiotic induction, but only in dbcAMP arrested oocytes. Purine de novo synthesis was nearly doubled in OCC by FSH treatment, and this response was completely prevented by Ado. FSH had no effect on HX salvage, although Ado reduced this activity by 98%. Inosine effects on metabolism were intermediate between the control and Ado groups. Experiments with radiolabeled energy substrates showed that Ado suppressed FSH activation of the pentose phosphate pathway but did not prevent significant activation of glycolysis or oxidation of pyruvate. Finally, in cultured follicles from primed mice, hCG-induced maturation was blocked by Ado as effectively as by the purine de novo synthesis inhibitor, azaserine. It is concluded that Ado has an inhibitory action on hormone-induced maturation that is due, at least in part, to suppression of glucose metabolism, leading to compromised purine de novo synthesis. PMID- 10813850 TI - Relationship between the association of rat epididymal protein "DE" with spermatozoa and the behavior and function of the protein. AB - Rat epididymal glycoprotein DE associates with the dorsal region of the sperm head during sperm maturation, migrates to the equatorial segment (ES) with the acrosome reaction (AR), and is involved in gamete membrane fusion. In the present study we examined the association of DE with the sperm surface and the relationship of this interaction with the behavior and function of the protein. Cloning and sequencing of DE revealed a lack of hydrophobic domains and the presence of 16 cysteine residues in the molecule. Experiments in which cauda epididymal sperm were subjected to different extraction procedures indicated that while most of the protein is removable from sperm by mild ionic strength, a low amount of DE, resistant to even 2 M NaCl, can be completely extracted by agents that remove integral proteins. However, the lack of hydrophobic domains in the molecule and the failure of DE to interact with liposomes, does not support a direct insertion of the protein into the lipid bilayer. These results, and the complete extraction of the tightly bound protein by dithiothreitol, suggest that this population would correspond to a peripheral protein bound to a membrane component by strong noncovalent interactions that involve disulfide bonds. While ELISA experiments showed that no protein could be extracted by NaCl from capacitated sperm, indirect immunofluorescence studies revealed the ability of the NaCl-resistant protein to migrate to the ES. Together, these results support the existence of two populations of DE: a major, loosely bound population that is released during capacitation, and a minor strongly bound population that remains after capacitation, migrates to the ES with the AR, and thus would correspond to the one with a role in gamete fusion. PMID- 10813851 TI - Expression of alpha and gamma interferon receptors in the sperm cell. AB - The present study was conducted to examine whether or not the sperm cell has the expression of receptors for interferon (IFN) -alpha and -gamma. This was investigated using specific antibodies. Antibody to IFN-alpha receptor reacted with the acrosomal and tail regions of the murine sperm cell in the indirect immunofluorescence technique (IFT) and immunoscanning electron microscopic procedure (ISEP). In the immunoprecipitation and Western blot procedures, this antibody specifically recognized a protein band of approximately 100 kD, which corresponds to the molecular weight of IFN-alpha receptor present in other cell types. Antibody to IFN-gamma receptor specifically reacted with the posterior head, midpiece, and tail regions of sperm cell in IFT and ISEP, and recognized a band of approximately 85 kD in the immunoprecipitation and Western blot procedures, corresponding to the IFN-alpha receptor. Similar bands of approximately 100 kD and approximately 85 kD molecular identities were also detected in the testes extracts and sperm extracts of other mammalian species namely human, rabbit, and pig, the species tested. These findings indicate that the mammalian sperm cell has expression of IFN-alpha and IFN-gamma receptors, which seem to develop during spermatogenesis in the testes. These findings may have implications in male infertility and antisperm contraceptive vaccine development. PMID- 10813852 TI - New site of temporary storage of rRNA in O. mykiss previtellogenic oocytes. AB - This article describes a new organelle found in the cytoplasm of the growth stage fish oocytes. In particular, we describe its organization at the morphological level and investigate its composition by different cytochemical and immunocytochemical approaches with both light and electron microscope. The conclusion is that the body is a peculiar protein scaffold functioning as a temporary trap for the storage of rRNA in the mid to late growth stage oocytes. Its presence would be related to the reorganization of the mass of amplified rDNA in micronucleoli and to the consequent temporary stop in the rRNA synthesis. PMID- 10813853 TI - Evidence for a functional glycogen metabolism in mature mammalian spermatozoa. AB - The glycogen content in fresh raw dog spermatozoa was 0.22+/-0.03 micromol/mg protein. This matched with the presence of a glycogen-like staining in the head and midpiece. Glycogen levels lowered to 0.05 micromol/mg protein after incubation for 60 min without sugars. Addition of either 10 mM fructose or 10 mM glucose increased glycogen content to 0.70 micromol/mg protein. On the other hand, glycogen synthase activity ratio of fresh dog sperm (0.35+/-0.07, measured in the absence and the presence of glucose 6-P) increased to 0.55 with 10 mM fructose for 20 min, whereas glucose had a smaller effect. Spermatozoa extracts had also a protein of about 100 Kd, which reacted against a rat liver glycogen synthase antibody. This was located in sperm head and midpiece. Furthermore, glycogen phosphorylase activity ratio measured in presence and absence of AMP (0.25+/-0.03 in fresh samples) decreased to 0.15 by 10 mM glucose for 20 min, whereas fructose was less potent in this regard. The maximal effect of glucose and fructose were observed from 10-20 mM onwards. This work is the first indication for a functional glycogen metabolism in mammal spermatozoa, which could play an important role in regulating sperm survival in vivo. PMID- 10813855 TI - Right ventricular function in patients after acute myocardial infarction assessed with phase contrast MR velocity mapping encoded in three directions. AB - The purpose of the study was to assess global and regional myocardial function of the right ventricle (RV) with the use of phase contrast (PC) velocity mapping in patients after acute myocardial infarction. We examined 8 patients after acute myocardial wall infarction and 10 healthy volunteers for comparison. PC velocity mapping was performed in a single midventricular short-axis slice with velocity encoding in three different directions. RV displacement during systole in the through-plane direction differed significantly between patients and volunteers (P = 0.009). RV myocardial velocity in the through-plane and radial directions, evaluated at time of peak ejection rate, was significantly lower in patients than in healthy volunteers (P<0.05). RV abnormalities may be detected in patients after acute myocardial infarction using PC velocity mapping with velocity encoding in three different directions. Owing to their short acquisition times and relatively easy postprocessing, PC techniques are time-efficient and promising tools for the evaluation of RV function. PMID- 10813854 TI - Transforming growth factor-betas in pre-gastrulation development of mammals (minireview). PMID- 10813856 TI - Application of breath-hold T2-weighted, first-pass perfusion and gadolinium enhanced T1-weighted MR imaging for assessment of myocardial viability in a pig model. AB - The purpose of this study was to correlate the abnormal signal area on various magnetic resonance (MR) images to the infarct area on pathologic examination and to assess the myocardial viability on the basis of MR images. T2-weighted, first pass perfusion, and delayed gadolinium-enhanced T1-weighted images were used as "one-stop examinations" in a pig model of reperfused myocardial infarction. The results of each MR image were compared with those of 2,3, 5-triphenyltetrazolium chloride (TTC) staining. The abnormal signal areas on T2-weighted and Gd-enhanced T1-weighted images were larger than the infarct areas on TTC staining (34.7% and 32.3% vs. 28.3%; P< 0.05), whereas the nonperfused areas on perfusion images were correlated (25.6% vs, 28.3%; P = 0.139). Electron microscopic examination showed severely distorted ultrastructures in the infarct areas and mildly damaged ultrastructures in the peri-infarct areas. Perfusion images probably reflected the infarct areas, whereas T2-weighted and Gd-enhanced T1-weighted images seemed to include peri-infarct as well as infarct areas. PMID- 10813857 TI - Takayasu arteritis: diagnosis with breath-hold contrast-enhanced three dimensional MR angiography. AB - The purpose of the study was to determine the diagnostic accuracy of breath-hold contrast-enhanced three-dimensional (3D) magnetic resonance (MR) angiography in Takayasu arteritis. Thirty patients suspected of having Takayasu arteritis were examined with MR angiography and conventional angiography. Takayasu arteritis was diagnosed in 20 of these patients. MR angiography was performed using a 1.5-T system after bolus injection of 0.1 mmol/kg of gadodiamide. MR angiography clearly depicted various vascular lesions in the aorta and its major branches in all 20 patients with Takayasu arteritis. It also depicted pulmonary artery lesions in 10 (50%) of the 20 patients. MR angiography accurately depicted 323 (98%) of 330 arteries, but 7 (2%) stenotic arteries were overestimated as occluded. The sensitivity and specificity of MR angiography for the diagnosis of Takayasu arteritis were both 100%. Breath-hold contrast-enhanced 3D MR angiography clearly depicts various vascular lesions in both the systemic and pulmonary arteries in Takayasu arteritis, thus allowing a definitive diagnosis of Takayasu arteritis. PMID- 10813858 TI - NC100150 Injection, a preparation of optimized iron oxide nanoparticles for positive-contrast MR angiography. AB - A preparation of monocrystalline iron oxide nanoparticles with an oxidized starch coating, currently in clinical trials (NC100150 Injection; CLARISCAN), was characterized by magnetization measurements, relaxometry, and photon correlation spectroscopy. By combining the results with a measure of iron content, one can obtain the size and magnetic attributes of the iron cores, including the relevant correlation times for outer sphere relaxation (tau(SO) and tau(D)), and information about the interaction of the organic coating with both core and solvent. The results are 6.43 nm for the iron oxide core diameter, a magnetic moment of 4.38x10(-17) erg/G, and a water-penetrable coating region of oxidized oligomeric starch fragments and entrained water molecules. The latter extends the hydrodynamic diameter to 11.9 nm and lowers the average diffusivity of solvent about 64% (which increases tau(D) accordingly). The nanoparticles show little size-polydispersity, evidenced by the lowest value of r(2)/r(1) at 20 MHz reported to date, an asset for magnetic resonance angiography. PMID- 10813859 TI - Dynamic susceptibility contrast MR imaging of plaque development in multiple sclerosis: application of an extended blood-brain barrier leakage correction. AB - Since the pathogenesis of multiple sclerosis (MS) lesions is not yet fully understood, we investigated the potential of dynamic susceptibility contrast (DSC) magnetic resonance (MR) perfusion imaging for a better characterization of lesion pathology. Twenty-five MS patients were examined on a 1.5 T scanner. A single dose of gadolinium (Gd)-DOTA contrast agent was injected, and echoplanar images were acquired every 0.5 seconds for 1 minute. From the signal intensity versus-time curves, the relative cerebral blood volume (rCBV) was evaluated for regions in plaques and in gray and white matter. The rCBV calculated for acute, Gd-enhancing plaques was corrected for the effects of blood-brain barrier leakage, using a new correction algorithm. Acute plaques had significantly higher blood volumes than normal-appearing white matter (P < = 0.01). Chronic plaques that appeared hypointense on T(1)-weighted images had lower rCBV than T(1) isointense plaques (P < = 0.03). Our results indicate that the acute phase in MS is accompanied by vasodilation. In later stages of gliosis, the perfusion decreases with increasing axonal injury. Although the DSC technique is less sensitive than conventional MR imaging, the information provided is essentially different from that obtained with any other MR method. PMID- 10813860 TI - Assessment of cerebral gliomas by a new dark fluid sequence, high intensity REduction (HIRE): a preliminary study. AB - The purpose of this study was to assess the diagnostic potential of a new dark fluid sequence, high intensity reduction (HIRE) in the diagnostic workup of patients with cerebral gliomas. The HIRE sequence utilizes a very long T(2) value of the cerebrospinal fluid (CSF) to suppress its high signal contribution in T(2) weighted imaging by a image subtraction technique. Fifteen patients with histologically confirmed cerebral gliomas were examined with T(2)-weighted fast spin-echo (FSE), T(1)-weighted SE, fast fluid-attenuated inversion recovery (FLAIR), and HIRE imaging using identical scan parameters. In patients with enhancing lesions, fast FLAIR and HIRE were added to the contrast-enhanced T(1) weighted SE images. Images were analyzed in a qualitative and quantitative evaluation. In the qualitative analysis, lesion detection, lesion delineation, and differentiation between enhancing and non-enhancing tumor tissue were assessed in a two-reader study. For the quantitative analysis, lesion-to background and lesion-to-CSF contrast and contrast-to-noise ratios were determined in a region of interest analysis. HIRE achieved a significant reduction of the CSF signal without losing the high gray-to-white matter contrast of T(2)-weighted sequences. In the quantitative analysis, the contrast ratios of the HIRE images were lower compared with the FLAIR images due to a relatively high background and CSF signal. After administration of contrast media, HIRE images presented a significant signal increase in enhancing lesions, which subsequently increased the contrast and contrast-to-noise ratios. In the qualitative analysis, both readers found all tumors clearly delineated on HIRE imaging. Compared with T(2)-weighted FSE, the tumor delineation with HIRE was better in nine patients, equal in four patients, and less in one patient. Compared with the FLAIR images, HIRE was rated superior in three patients, equal in nine patients, and inferior in another three patients. Delineation of the enhancing tumor parts was possible with HIRE in all patients. HIRE images had significantly fewer image artifacts than FLAIR images due to reduced inflow effects. The T(2)-based HIRE sequence presented is an alternative to the T(1) based FLAIR sequence, with the advantage of better gray-to-white matter contrast and shorter measurement time. Due to the subtraction technique, signal intensities from tissues with relaxation times in the range T(2 WM) < < T(2) < T(2 CSF) are also gradually affected, corresponding to their T(2) values. With respect to this unwanted effect, an improvement in HIRE imaging will be possible by using a self-weighted subtraction algorithm. In a forthcoming study this concept will first be tested on appropriate phantom fluids. PMID- 10813862 TI - An image processing strategy for the quantification and visualization of exercise induced muscle MRI signal enhancement. AB - Exercise increases the skeletal muscle water signal in T2-weighted images. Potential medical applications of MR studies of exercise-induced muscle signal intensity changes are the assessment of myopathies, sport training regimens, and physical therapy approaches following surgeries. We developed an automated image processing technique that provides volumetric analysis and visualization of exercise-related T2-weighted image intensity changes. The image processing was applied to the segmentation and quantification of activated muscle volumes. Qualitative assessment of muscle activation is demonstrated with three dimensional surface rendering. Quantitative determination of active muscle volume, signal intensity, and change over time is demonstrated. Visualization of the activated muscles allows functional anatomical assessment of exercise, which in turn allows detection of muscle utilization. PMID- 10813861 TI - Prospective comparison of magnetic resonance sialography and digital subtraction sialography. AB - We sought to compare the diagnostic utility of magnetic resonance sialography (MRS) and digital subtraction sialography (DSS) in patients with suspected sialolithiasis or sialadenitis. Sixteen consecutive patients (4 female and 12 male, mean age 51+/-16 years) with suspected sialolithiasis or sialadenitis underwent DSS by a standard technique and MRS. MRS was obtained with a T2 weighted single-shot TSE sequence (TR/TE 2800/1100 msec, acquisition time 7 seconds) using a quadrature head (n = 16) and a surface coil (n = 8). Nineteen symptomatic glands were investigated with DSS: eight submandibular glands in 6 patients (two bilateral) and unilateral parotid glands in 11 patients. MRS was always carried out to visualize gland ducts bilaterally. The ductal system was visualized in all glands examined by MRS. DSS depicted the ductal system in all 11 parotid glands, but only 4 of the 8 submandibular glands (50%). Sialolithiasis was diagnosed in three cases (one parotid, two submandibular glands) by MRS and in two cases by DSS. DSS demonstrated tertiary branching ducts and MRS secondary branching ducts. MRS is able to visualize the ductal system of the parotid and submandibular gland noninvasively and is thus not dependent on successful cannulation of the orifice of the ductal system. Our preliminary data indicate that MRS is useful for diagnosing sialolithiasis. MRS allows diagnosis of sialadenitic changes, but DSS achieves a better diagnostic performance due to higher spatial resolution. PMID- 10813863 TI - Effect of exercise on (23)Na MRI and relaxation characteristics of the human calf muscle. AB - The acute affect of voluntary muscle contractions performed by healthy volunteers was evaluated using (23)Na nuclear magnetic resonance (NMR). Three-dimensional gradient-echo (23)Na images, pulse-acquired spectra, and transverse relaxation times were obtained before and after ankle flexion-extension exercise. The muscle sodium concentration was calculated from (23)Na images using a 40 mM NaCl standard and the measured T(2) values. Before exercise the muscle sodium concentration was 26+/-4 mmole/kg wet weight. This agrees closely with literature values, suggesting that muscle Na(+) is fully NMR visible. The (23)Na image intensity increased by 34%+/-7% in the exercised muscle and diminished with a half-life of 30+/-6 minutes. The pulse-acquired spectra, however, did not show any significant change in muscle signal intensity following exercise, but the relative contribution of the slow T(2) component increased. The calculated sodium concentration also did not change significantly after the exercise. We therefore infer that the changes in (23)Na magnetic resonance imaging (MRI) were due to a change in sodium-macromolecular interaction rather than a change in tissue sodium content. We believe that this report represents the first study of (23)Na MRI of skeletal muscle. PMID- 10813864 TI - Optimum setting of binomial pulses for magnetization transfer contrast. AB - Investigation of the cause of image artifacts generated by a magnetization transfer (MT) sequence using binomial-pulse trains led to findings of imperfections in the pulses. These imperfections caused anomalous direct saturation of the free water, which was localized due to the static magnetic field inhomogeneity. In the case of single binomial pulses a loss of overall MT response across the field of view results. Two methods of correcting the imperfections and removing the artifact have been established using interactive adjustment of sub-pulse lobes and phase swapping of pulse trains. These imperfections may be present in many systems and may have led to erroneous judgements of the value of binomial pulses for MT imaging. A technique for interrogating the frequency spectrum of the binomial-pulse train has been utilized, allowing its optimization. The use of accurate and optimized binomial pulses may yet prove to be preferable to pulsed off-resonance methods for quantitative, clinical MT imaging. PMID- 10813865 TI - A generalized k-sampling scheme for 3D fast spin echo. AB - The phase-encoding scheme can significantly affect the quality of fast spin-echo (FSE) images because the echo amplitude is modulated as a function of the echo position in k-space. The effects of the modulation in two-dimensional FSE imaging include ghosting and blurring artifacts and resolution loss in the phase-encoding (PE) direction. In 3D FSE imaging, the use of two PE directions presents the opportunity for improved PE schemes. A new scheme for assignment of echoes to views in 3D FSE, termed generalized, has been developed. This scheme distributes T(2) effects along both PE directions, allowing considerable flexibility in the selection of blurring artifact appearance. In a set of simulations, phantom experiments, and in vivo experiments, the performance of the generalized PE scheme for 3D FSE imaging was compared with the performance of existing PE schemes. The results demonstrate that the generalized PE scheme can be used to reduce blurring artifacts greatly relative to other PE techniques that are presently in use. This approach to PE can be used to manipulate the blurring artifact appearance and to optimize acquisition time. PMID- 10813866 TI - Differences in water diffusion and lactate production in two different types of postinfectious encephalopathy. AB - We compared two different types of postinfectious encephalopathy using magnetic resonance imaging (MRI), including diffusion-weighted images and MR spectroscopy. Acute disseminated encephalomyelitis (ADEM) and acute necrotizing encephalopathy (ANE) showed different distribution of abnormal intensity areas and different diffusion of water measured by diffusion-weighted imaging (DWI). Proton MR spectroscopy (MRS) showed lactate production in both cases, which returned to a normal range; the rate of increased lactate production was much lower in the ANE case. Water diffusion showed a difference in pathophysiological background between the two encephalopathies, but the lactate elevation observed by proton MRS did not correlate with clinical severity. PMID- 10813867 TI - An integrated head-holder/coil for intraoperative MRI in open neurosurgery. AB - With the invention of "open" magnetic resonance imaging (MRI) systems, access to the patient is possible during the imaging procedure. An important application of these systems is intraoperative MRI to control the extent of resection during tumor surgery. Up to now flexible surface coils wrapped around, or placed at each side of the head, were used for imaging. These flexible coils have several disadvantages such as unreliability, interindividual problems, difficult handling, poor hygienic properties, and often unsatisfactory or inhomogeneous image quality. To solve most of these problems, an MR-compatible head-holder in combination with an integrated surface coil for use in a 0.2 T C-shaped magnet was developed. Forty-eight patients with known cranial tumors underwent MRI intraoperatively. In 32 patients (67%), residual tumor was found, and additional surgical resection was performed. The integrated head-holder/coil is a safe and practical tool for intraoperative MRI, providing efficient and reliable resection control during neurosurgical procedures. PMID- 10813868 TI - A multicenter measurement of magnetization transfer ratio in normal white matter. PMID- 10813870 TI - Reaction kinetics at very low temperatures: laboratory studies and interstellar chemistry. AB - Studies of gas-phase processes at temperatures down to 10 K have recently blossomed, largely through application of the CRESU (cinetique de reaction en ecoulement supersonique uniforme) technique. The results are of considerable relevance to the synthesis of molecules in dense interstellar clouds, demonstrating that the models developed to explain the observed molecular abundances must be expanded to include reactions between electrically neutral species. In addition, the experimental results have stimulated theoretical efforts to describe the factors that control the rates of such low-temperature reactions. In this Account, the CRESU method is described and the relevance of the results discussed. PMID- 10813869 TI - Increased expression of N-myristoyltransferase in gallbladder carcinomas. AB - BACKGROUND: Activated Src, which has intrinsic protein tyrosine kinase activity, has been found in human solid tumors such as colorectal and breast carcinomas. The Src gene encodes a cytoplasmic tyrosine kinase p60src, which attaches to the inner surface of the membrane after N-terminal myristoylation and is implicated in transduction of signals to the nucleus. N-myristoyltransferase (NMT) catalyzes the biochemical modification process called N-myristoylation. To investigate whether, through Src, NMT contributes to the pathogenesis of gallbladder carcinoma, the authors investigated expression of NMT and p53 in in situ and invasive carcinomas. METHODS: One hundred cases of documented gallbladder carcinoma were reviewed, and 30 cases were selected randomly to evaluate expression of NMT and p53 by immunohistochemistry in both in situ and in invasive tumor components. RESULTS: Eighteen cases (60%) of gallbladder carcinoma showed moderate to strong cytoplasmic positivity for NMT with increased intensity in the invasive component, and 12 cases (40%) were negative. The in situ component revealed mild to moderate cytoplasmic staining in 20 cases (67%), whereas the normal gallbladder mucosa showed weak to negative cytoplasmic staining. Moderate to strong p53 staining was observed in 17 in situ cases (63%) and 24 invasive cases (80%). The in situ staining patterns of p53 were unrelated to the clinical outcome of the tumor. However, moderate to strong staining of the invasive component as observed in 15 cases (50%) was associated with a mean survival of 8.8 months. Amplification of intron-8 in normal gallbladder mucosa and invasive carcinoma were similar in intensity, suggesting the absence of NMT gene amplification in these tumors. CONCLUSIONS: The increased expression of NMT in these tumors could be due to transcriptional activation. Tumors with increased expression of NMT and p53 were associated with poor clinical outcomes as evidenced by their mean survival times. NMT is likely to play a pathogenic role in gallbladder carcinoma. PMID- 10813871 TI - Molecular photovoltaics. AB - The dye-sensitized nanocrystalline injection solar cell employs transition metal complexes for spectral sensitization of mesoporous TiO(2) films together with suitable redox electrolytes or amorphous organic hole conductors. Light harvesting occurs efficiently over the whole visible and near-IR range due to the very large internal surface area of the films. Judicious molecular engineering allows the photoinduced charge separation to occur quantitatively within a few femtoseconds. The certified overall power conversion efficiency of the new solar cell for AM 1.5 solar radiation stands presently at 10.4%. PMID- 10813872 TI - Stereoelectronic control in Diels-Alder reaction of dissymmetric 1, 3-dienes. AB - The Diels-Alder reaction is a widely employed protocol in which four stereogenic centers are generated in a predictable manner with olefin geometry, adjoining chiral center, and transition-state topology serving as the main controlling elements. However, when the Diels-Alder partners are in a dissymmetric environment, pi-face selection is determined through the interplay of steric, orbital, and electrostatic factors whose relative importance is a subject of intense debate. Several new systems have been crafted to probe the mechanistic nuances of the pi-face selection. The available data have enabled us to qualitatively define a hierarchy of various stereoelectronic effects that would aid predictability of the stereochemical outcome. PMID- 10813873 TI - Electron and proton reservoir complexes: thermodynamic basis for C-H activation and applications in redox and dendrimer chemistry. AB - This Account focuses on fundamental aspects and applications of the concept of "electron and proton reservoirs". Permethylated electron reservoir iron sandwich complexes, which were known to be excellent reducing agents in their neutral 19e Fe(I) form, have recently been shown to be stable strong oxidants in their dicationic 17e Fe(III) form. Thermodynamic, mechanistic, and synthetic aspects of the redox chemistry of these and related organoiron complexes are summarized. Proton reservoir properties of these complexes are due to enhanced acidity in their cationic form and result in an original type of activation: perfunctionalization leading to dendritic cores and useful metallodendrimers of variable topology. PMID- 10813874 TI - Identification of peptide oxidation by tandem mass spectrometry. AB - Oxidative reactions play important roles in a variety of biochemical events ranging from normal metabolism to aging and disease processes. Proteins represent major targets for modification in these reactions, and identification of sites and structures of modifications may lead to mechanistic understanding and approaches for prevention. In this Account, the utility of mass spectrometry and its advantages are described for the identification of oxidative modifications to protein targets. A variety of examples are provided to illustrate how modifications are accurately identified and quantitated using modern methods of ionization coupled with HPLC and tandem mass spectrometry. PMID- 10813875 TI - Spectroscopic and dynamic studies of the epidermal chromophores trans-urocanic acid and eumelanin. AB - Photoaging and skin cancer can result from the exposure of skin to ultraviolet A (UV-A, 320-400 nm) radiation. The detailed chemical mechanisms by which these processes occur are not known, but they must begin with the absorption of a UV-A photon by one or more photoreceptor(s) within the skin. The situation is complicated by the lack of understanding of the photoreactions of many of the UV A-absorbing molecules in skin. In this Account, we describe recent research efforts directed at elucidating the UV-A-induced photoreactivity of two light absorbing epidermal photoreceptors: trans-urocanic acid and eumelanin. PMID- 10813876 TI - Anionic Pd(0) and Pd(II) intermediates in palladium-catalyzed Heck and cross coupling reactions. AB - The anions of PdCl(2)L(2) and Pd(OAc)(2), precursors of palladium(0) used in cross-coupling and Heck reactions, play a crucial role in these reactions. Tricoordinated anionic complexes Pd(0)L(2)Cl(-) and Pd(0)L(2)(OAc)(-) are the effective catalysts instead of the usually postulated Pd(0)L(2) complex. The anion ligated to the palladium(0) affects the kinetics of the oxidative addition to ArI as well as the structure and reactivity of the arylpalladium(II) complexes produced in this reaction. Thus, pentacoordinated anionic complexes are formed, ArPdI(Cl)L(2)(-) or ArPdI(OAc)L(2)(-), the precursor of neutral trans ArPd(OAc)L(2), instead of the usually postulated trans-ArPdIL(2) complex (L = PPh(3)). PMID- 10813877 TI - Journal of Combinatorial Chemistry: Our Next Millennium. PMID- 10813878 TI - Definition of Templates within Combinatorial Libraries. PMID- 10813879 TI - A Method for Quantitation of Solid-Phase Synthesis Using (19)F NMR Spectroscopy. PMID- 10813880 TI - Parallel Synthesis of Alkyl Tetrazole Derivatives Using Solid Support Chemistry. AB - The synthesis of several omega-chloroalkyl tetrazoles and their subsequent attachment to a solid support is described. Using an in situ Finkelstein reaction, a variety of nucleophiles were alkylated and then cleaved from the resin to give pure alkyl tetrazole derivatives. A sample library of 5 x 6 demonstrates the general utility of this sequence. PMID- 10813881 TI - N-Arylation of Sulfonamides on Solid Supports. AB - A general and mild method for the N-arylation of sulfonamides on solid supports is reported. Copper acetate, triethylamine mediated coupling of arylboronic acids at room temperature to solid-supported sulfonamides gave good to excellent yields of the desired N-arylsulfonamides. Sulfonamide bond cleavage of the o,p dinitrobenzene(N-aryl)sulfonamide provides a route to N-arylated secondary amine products. PMID- 10813882 TI - On-Bead Quantitation of Resin Bound Functional Groups Using Analogue Techniques with Vibrational Spectroscopy. AB - This report introduces two novel methodologies for the on-bead, functional group quantitation of solid phase synthesis using vibrational spectroscopy. "Analogue bleed" quantitation is an infrared spectroscopic technique in which combinatorial library bead spectral data are regressed against solution phase functional analogue spectra of a ligand in question. "Dual Analogue" quantitation is introduced as an infrared or Raman technique in which ligand-to-styrene band ratios of solution phase ligand/styrene monomer standards are used to generate an internally path length referenced calibration which can then be applied to solvent swollen beads. These technologies have application across a wide range of functional groups and can be applied throughout each step of a resin bound synthesis. PMID- 10813883 TI - 2,5-Dimethylfuran (DMFu): An Internal Standard for the "Traceless" Quantitation of Unknown Samples via (1)H NMR. AB - The use of 2,5-dimethylfuran (DMFu) as an internal standard for the quantitation of unknown samples by (1)H NMR is described. DMFu was selected based upon its low chemical reactivity, good solubility in standard NMR solvents, appropriate volatility (bp 92 degrees C), and the chemical shifts of its two singlets (5.80 and 2.20 ppm). In particular, the widely separated singlets of DMFu offer two advantages over more traditional "single peak" internal standards (e.g., hexamethyldisiloxane): (1) they reduce the possibility for overlap between analyte and internal standard signals; and (2) they enable the chemist to assess the accuracy of integration (and ergo, the quantitation) by calculating the ratio of the two singlets and comparing the observed ratio to the theoretical value. Our experiments have shown that the precision of the ratio of DMFu singlets strongly correlates to the precision of sample quantitation, an observation which greatly simplifies the assessment of the quality of the quantitation. Utilizing this technique, we have developed straightforward conditions for (1)H NMR experiments which consistently provide quantitative results with less than 5.0% error. PMID- 10813884 TI - A Versatile Approach to the Solution-Phase Combinatorial Synthesis of Substituted Pyridines: The Cobalt-Catalyzed Cyclotrimerization of Alkynes with a Nitrile. AB - The cobalt-catalyzed trimerization of two alkynes and one nitrile was exploited to produce solution-phase libraries of pyridines. Upon addition of 2 equiv of a CO scavenger (i.e., an amine-N-oxide), the commercially available [CpCo(CO)(2)] proved to be an attractive catalyst, as the required reaction temperatures, side product formation (i.e., carbocycles), and oxygen sensitivity are moderate. Purification of the reaction mixture was performed with an acidic cation-exchange resin. PMID- 10813885 TI - Kinetic Comparison of Trifluoroacetic Acid Cleavage Reactions of Resin-Bound Carbamates, Ureas, Secondary Amides, and Sulfonamides from Benzyl-, Benzhydryl-, and Indole-Based Linkers. AB - The kinetics of cleavage reactions of 16 resin-bound carbamates, ureas, secondary amides, and sulfonamides from four different acid labile linkers including benzyl, benzhydryl, and indole linkers has been investigated. The optimized cleavage conditions are generally milder than those commonly used and reported (e.g., 0.5% TFA as opposed to 5%). Among various linkers studied in this work, the indole linker has been found to be the most acid labile followed by the Rink linker. The rate of cleavage of compounds linked to the resin via various functional groups can be summarized as follows: sulfonamide >carbamate approximately urea > amide. This study shows that cleavages of 16 compounds from four different acid labile linkers have been optimized to much milder conditions in terms of TFA concentration and the reaction time. It also demonstrates that single bead FTIR is an effective tool for optimizing cleavage conditions. PMID- 10813887 TI - Editorial PMID- 10813886 TI - Preparation of Designer Resins via Living Free Radical Polymerization of Functional Monomers on Solid Support. AB - Merrifield resin is converted to a solid-supported free radical initiator by reacting with the TEMPO-Na. Heating TEMPO-methyl resin with a variety of functionalized styrene and acrylate monomers gives larger resin beads via living free radical polymerization. We have coined the term Rasta resin to describe resin beads prepared in this fashion. The process can be described as a solvent free suspension polymerization. It is particularly well suited for preparation of resin beads from monomers which contain electrophilic groups that would be destroyed upon suspension polymerization in water. Rasta resins have a novel macromolecular architecture wherein long straight chain polymers bearing reactive functional groups emanate from the phenyl groups of a cross-linked polystyrene core. With judicious choice of co-monomers and polymerization strategy, the solvent affinity, loading capacity, and distance of functionality from the cross linked core may be controlled giving beads with properties that are tailored to specific uses as synthesis supports and scavenging resins. PMID- 10813888 TI - Flexible synthesis and biological activity of uronic acid-type gem-diamine 1-N iminosugars: a new family of glycosidase inhibitors. AB - An efficient and flexible synthetic route to four gem-diamine 1-N-iminosugars of uronic acid-type (D-glucuronic, D-mannuronic, L-iduronic, and L-guluronic acid), a new family of glycosidase inhibitor, from l-galactono-1,4-lactone have been developed in an enantiodivergent fashion through a sequence involving as the key steps (a) the formation of gem-diamine 1-N-iminopyranose ring by the Mitsunobu reaction of an aminal and (b) the introduction of a carboxylic acid group by the Wittig reaction of a ketone, hydroboration and oxidation, and the Sharpless oxidation. D-Glucuronic and D-mannuronic acid-type 1-N-iminosugars, (3S,4R,5R, 6R)- and (3S,4R,5R,6S)-4, 5-dihydroxy-6-trifluoroacetamido-3-piperidinecarboxylic acid, were proven to be potent inhibitors for beta-D-glucuronidase (IC(50) 6.5 x 10(-)(8)M) and to affect human heparanase (endo-beta-glucuronidase). PMID- 10813889 TI - Pyridinolysis of phenyl-substituted phenyl chlorophosphates in acetonitrile AB - The kinetics and mechanism of the reactions of phenyl-substituted phenyl chlorophosphates, I, with pyridines in acetonitrile are investigated at 25.0 degrees C. The rates are much faster and the transition state is much earlier on the reaction coordinate with smaller beta(X) (0.16-0.18) and smaller negative beta(XY) (-0.011) values than those for the corresponding reactions with anilines. The vertical approach of the pyridine ring to an apical position of the trigonal bipyramidal transition state enables the p(pi)-d(pi) overlap between the pi orbital of pyridine and an empty d-orbital of phosphorus with less steric hindrance in a concerted process. The activation parameters are in line with the proposed mechanism. PMID- 10813890 TI - Thermal decomposition mechanisms of tert-alkyl peroxypivalates studied by the nitroxide radical trapping technique AB - The thermolysis of a series of tert-alkyl peroxypivalates 1 in cumene has been investigated by using the nitroxide radical-trapping technique. tert-Alkoxyl radicals generated from the thermolysis underwent the unimolecular reactions, beta-scission, and 1,5-H shift, competing with hydrogen abstraction from cumene. The absolute rate constants for beta-scission of tert-alkoxyl radicals, which vary over 4 orders of magnitude, indicate the vastly different behavior of alkoxyl radicals. However, the radical generation efficiencies of 1 varied only slightly, from 53 (R = Me) to 63% (R = Bu(t)()), supporting a mechanism involving concerted two-bond scission within the solvent cage to generate the tert-butyl radical, CO(2), and an alkoxyl radical. The thermolysis rate constants of tert alkyl peroxypivalates 1 were influenced by both inductive and steric effects [Taft-Ingold equation, log(rel k(d)) = (0.97 +/- 0. 14)Sigmasigma - (0.31 +/- 0.04)SigmaE(s)(c), was obtained]. PMID- 10813891 TI - Synthesis of novel donor mimetics of UDP-Gal, UDP-GlcNAc, and UDP-GalNAc as potential transferase inhibitors. AB - For the enzymatic transfer of galactose, N-acetylglucosamine, and N acetylgalactosamine, UDP-Gal (1), UDP-GlcNAc (2), and UDP-GalNAc (3) are employed, and UDP serves as a feedback inhibitor. In this paper the synthesis of the novel UDP-sugar analogues 4, 5, and 6 as potential transferase inhibitors is described. Compounds 4-6 feature C-glycosidic hydroxymethylene linkages between the sugar and nucleoside moieties in contrast to the anomeric oxygens in the natural derivatives 1-3. PMID- 10813892 TI - Photoinduced reactions of chloranil with 1,1-diarylethenes and product photochemistry-intramolecular AB - Photoinduced reactions of chloranil (CA) with 1,1-diarylethenes 1 [(p-X Ph)(2)C=CH(2), X = F, Cl, H, Me] in benzene afforded products 4-14, respectively, with the bicyclo[4.2.0]oct-3-ene-2,5-diones 4, the 6-diarylethenylcyclohexa-2,5 diene-1,4-diones 5, and 2,3,5, 6-tetrachlorohydroquinone 13 as the major primary products. The cyclobutane products 4 are formed via a triplet diradical intermediate without involvement of single electron transfer (SET) between the two reactants, while 5 is derived from a reaction sequence with initial SET interaction between (3)CA and the alkene. The 9-arylphenanthrene-1,4-diones 6 and its 10-hydroxy-derivatives 7 are secondary photochemical products derived from 5. The isomeric cage products 9-11 are formed from 4 via intramolecular benzene alkene [2 + 2] (ortho-)photocycloadditions induced by the triplet excited enedione moiety. The relative amount of the two groups of products (4 and its secondary products 9-11 via non-SET route vs 5 and its secondary products 6, 7, 8, 12, and 14 via SET route) shows a rather regular change, with the ratio of non SET route products gradually increasing with the increase in oxidation potential of the alkenes and in the positive free energy change for electron transfer (DeltaG(ET)) between (3)CA and the alkene, at the expense of the ratio of the products from the SET route. The competition between the SET and non-SET routes was also found to be drastically influenced by solvent polarity, with the SET pathways more favored in polar solvent. Photo-CIDNP investigations suggest the intermediacy of exciplexes or contact ion radical pairs in these reactions in benzene, while in acetonitrile, SET process led to the formation of CA(*)(-) and cation radical of the alkene in the form of solvent separated ion radical pairs and free ions. PMID- 10813893 TI - A strategy for the asymmetric aminohomologation of alpha, beta-dihydroxy aldehydes: application to the synthesis of the southwest tripeptide segment of echinocandin B. AB - The synthesis of the (2S,3S,4S)-3,4-dihydroxyhomotyrosine amino acid segment, present in echinocandin B, in its activated form ready for peptide coupling is described. The key steps of the approach are the enantioselective AD reaction of 4-methoxycinnamic acid methyl ester, a completely diastereoselective [2 + 2] hydroxyketene-imine cycloaddition, and the TEMPO-assisted cycloexpansion of the resulting 3-hydroxy beta-lactam to the corresponding alpha-amino acid N-carboxy anhydride (NCA). The smooth opening of the latter upon treatment with L Thr(OSi(t)BuPh(2))OMe and further acylation with the N-Cbz protected L-4-tert butyldiphenylsilyloxy proline rendered the southwest portion of echinocandin B. PMID- 10813894 TI - Ab initio and density functional calculations on the pericyclic vs pseudopericyclic mode of conjugated nitrile ylide 1, 5-electrocyclizations AB - Ab initio (MP2/6-311+G and MP4(SDTQ)/6-311+G//MP2/6-311+G) and density functional (B3LYP/6-311+G) calculations on the ring closure reactions of conjugated nitrile ylides 1a-e, 3, and 6 to the corresponding oxazoles 2a, 5, 7, and 8; thiazoles 2b and 4; imidazole 2c; and pyrroles 2d and 2e, respectively, are reported. Vinyl nitrile ylides 1d and 1e cyclize with a substantially higher barrier than nitrile ylides containing a heteroatom. Geometric features as well as electronic structures as obtained by NBO analysis are indicative of a pericyclic, monorotatory 1, 5-electrocyclization of 1d and 1e. For nitrile ylides where X = heteroatom, a pseudopericyclic heteroelectrocyclization pathway, characterized by in-plane attack of the heteroatom's lone pair at the nitrile ylide group, is found. For 3 and 6, where two different cyclization products are possible, the calculated barriers and reaction energies are in line with the experimentally observed direction of reaction. Vinyl nitrile ylides 1d and 1e are characterized by an allene, acyl substituted derivatives 1a, 1b, 3, and 6 by a propargyl type structure. The nitrogen derivative 1c represents an intermediate case. PMID- 10813895 TI - Hydrozirconation of stannylacetylenes. Synthesis and reactions of ketene Stannyl(Telluro) acetals AB - Stannylacetylenes 7a-e react with Cp(2)Zr(H)Cl in THF at room temperature to give the alpha-zirconated vinylstannane intermediates 8a-e, which subsequently react with butyltellurenyl bromide (2.0 equiv) to give exclusively ketene stannyl(telluro) acetals 6a-e of E configuration. Similar reactions were performed using phenylselenenyl bromide (2.0 equiv) as the electrophile, but a mixture of products was formed including the expected ketene stannyl(seleno) acetals 12. Otherwise, the use of 1.4 equiv of Cp(2)Zr(H)Cl and 1.0 equiv of PhSeBr results in the exclusive formation of 12, in good yields. Treatment of ketene stannyl(telluro) acetals with iodine or NBS followed by reductive dehalogenation results in the formation of 1-iodo-1-telluroalkenes 4a-e and 1 bromo-1-telluroalkenes 5a-e, respectively, with total retention of the configuration. PMID- 10813896 TI - Hydrozirconation of lithium alkynylselenolate anions. Generation and reactions of alpha-zirconated vinyl selenide intermediates AB - Lithium alkynylselenolate anions react completely with 1.0 equiv of Cp(2)Zr(H)Cl in THF at room temperature to give exclusively the alpha-zirconated vinylselenolate intermediates 23-27, which by treatment with an alkyl halide afforded the alpha-zirconated vinyl alkylselenide intermediates 29-33. Reaction of 29-33 with butyltellurenyl bromide results in the formation of ketene telluro(seleno) acetals 35-39 with total control of the regio- and stereochemistry. The synthetic utility of the ketene telluro(seleno) acetals obtained here was demonstrated by reaction of 36 with butyllithium. This promotes the exclusive and stereospecific removal of the tellurium moiety and enables formation of the corresponding selenium-containing allylic alcohol of type 44, alpha-(alkylseleno)-alpha,beta-unsaturated aldehyde 45, ester 46, or carboxylic acid 47, after reaction with different types of electrophiles. PMID- 10813897 TI - Enantioselective inclusion of methyl phenyl sulfoxides and benzyl methyl sulfoxides by (R)-phenylglycyl-(R)-phenylglycine and the crystal structures of the inclusion cavities AB - Crystalline (R)-phenylglycyl-(R)-phenylglycine [(R,R)-1] includes methyl phenyl sulfoxides (2 and 3) and benzyl methyl sulfoxides (4) with high enantioselectivity. The dipeptide exhibited different stereoselectivity depending on four structural isomers of methyl tolyl sulfoxide (C(8)H(10)OS): R for methyl 2-tolyl sulfoxide, S for methyl 3-tolyl sulfoxide, and racemic for methyl 4-tolyl sulfoxide. A structural isomer, benzyl methyl sulfoxide, was included in racemic form. Chlorophenyl methyl sulfoxides 3 (C(7)H(7)ClOS) with a similar volume showed the same enantioselectivity for their recognition. By single-crystal X-ray analyses of these inclusion compounds, it was elucidated that (R,R)-1 molecules self-assembled to form layer structures and included the sulfoxides between these layers and that the origin of the enantioselectivity based on chiral cavities was induced by conformation of the C-terminal phenyl group of the dipeptide. The relative position between the ammonio proton and the C-terminal phenyl group in one molecule of the dipeptide determined the stereochemistry of the methyl sulfinyl groups to be recognized. Various positional isomers of methyl xylyl sulfoxide having the formula of C(9)H(12)OS were subjected to the enantioselective inclusion by (R,R)-1 crystals and these results are also discussed. PMID- 10813898 TI - Rationalization of enantioselectivities in dialkylzinc additions to benzaldehyde catalyzed by fenchone derivatives. AB - Three (-)-fenchyl alcohol derivatives, ?(1R,2R,4S)-exo-(2-Ar)-1,3, 3 trimethylbicyclo[2.2.1] heptan-2-ol, Ar = o-anisyl (2), 2-N-methylimidazolyl (3), 2-N,N-dimethylbenzylamine (4)? were synthesized, characterized by X-ray analyses, and employed as precatalysts in diethyl zinc additions to benzaldehyde. Directions and relative degrees of enantioselectivities are rationalized by QM/MM ONIOM computations of mu-O transition structure models. Enantioselectivities arise from repulsive interactions between "transferring" or "passive" alkyl groups at the zinc centers and the substituents at donor groups or the bicyclo[2.2.1]heptane moieties. These results enable predictions for ligand tuning to improve catalyst efficiency of fenchone-based ligands in dialkylzinc additions to aldehydes. PMID- 10813899 TI - Convenient enzymatic resolution of alcohols using highly reactive, nonharmful acyl donors, 1-ethoxyvinyl esters AB - 1-Ethoxyvinyl esters 3, a new type of acyl donors for enzymatic resolution of racemic alcohols, were disclosed to be superior to the contemporary major reagents, vinyl esters 1 and isopropenyl esters 2. Three features of 3 are noticeable: (1) 3 generates ethyl acetate as a single coproduct, which does not affect any enzymes, while acetaldehyde liberated from 1 deactivates some kinds of lipases. (2) The reactivity of 3 was not less than that of 1 and much higher than that of 2, and the optical purity of the products was as high as that of 1 and 2. Especially, it was generally observed that 3 showed higher reactivity than 1 for reactions using Candida rugosa lipases, one of the most commonly employed lipases, having liberal applicability to substrates but sensitive to acetaldehyde. Twelve examples of the kinetic resolution of racemic secondary alcohols (5 and 10) and one desymmetrization of meso-alcohol 7 were presented employing the acetate 3a or the octanoate 3b and four types of lipases. (3) A one pot procedure for the preparation of 3 from the corresponding carboxylic acid and the subsequent enzymatic resolution of alcohols, which has not been reported using 1 or 2, was elucidated. The chemical and optical yields of the products by this procedure were similar to those obtained using isolated 3. PMID- 10813900 TI - An efficient synthesis of peri-hydroxy aromatic compounds via a strong-base induced AB - An efficient synthesis of peri-hydroxy aromatic compounds has been accomplished via a strong-base-induced [4+2] cycloaddition of homophthalic anhydrides with alpha-sulfinyl-substituted derivatives of enolizable enones. The unsubstituted enones did not undergo an efficient [4+2] cycloaddition reaction with homophthalic anhydrides, presumably due to their enolization under the basic reaction conditions. The sulfinyl group not only promotes the cycloaddition reaction but also undergoes in situ elimination under the reaction conditions to afford the peri-hydroxy aromatic compounds in a single step. The application of this methodology for the synthesis of a key intermediate of antitumor antibiotic fredericamycin A is described. PM3 calculations of various 2-substituted cyclopentenones as well as the mechanism of the cycloaddition are also discussed. PMID- 10813902 TI - Oxidation of symmetric disulfides with hydrogen peroxide catalyzed by Methyltrioxorhenium(VII) AB - Organic disulfides with both alkyl and aryl substituents are oxidized by hydrogen peroxide when CH(3)ReO(3) (MTO) is used as a catalyst. The first step of the reaction is complete usually in about an hour, at which point the thiosulfinate, RS(O)SR, can be detected in nearly quantitative yield. The thiosulfinate is then converted, also by MTO-catalyzed oxidation under these conditions, to the thiosulfonate and, over long periods, to sulfonic acids, RSO(3)H. In the absence of excess peroxide, RS(O)SR (R = p-tolyl), underwent disproportionation to RS(O)(2)SR and RSSR. Kinetics studies of the first oxidation reaction established that two peroxorhenium compounds are the active forms of the catalyst, CH(3)ReO(2)(eta(2)-O(2)) (A) and CH(3)ReO(eta(2)-O(2))(2).(OH(2)) (B). Their reactivities are similar; typical rate constants (L mol(-)(1) s(-)(1), 25 degrees C, aqueous acetonitrile) are k(A) = 22, k(B) = 150 (Bu(2)S(2)) and k(A) = 1.4, k(B) = 11 (Tol(2)S(2)). An analysis of the data for (p-XC(6)H(4))(2)S(2) by a plot of log k(B) against the Hammett sigma constant gave rho = -1.89, supporting a mechanism in which the electron-rich sulfur attacks a peroxo oxygen of intermediates A and B. PMID- 10813901 TI - N-Alkyl-N-cyclopropylanilines as mechanistic probes in the nitrosation of N,N dialkyl aromatic amines. AB - A group of N-cyclopropyl-N-alkylanilines has been synthesized, and their reaction with nitrous acid in aqueous acetic acid at 0 degrees C was examined. All compounds reacted rapidly to produce the corresponding N-alkyl-N-nitrosoaniline by specific cleavage of the cyclopropyl group from the nitrogen. The transformations were unaffected by the nature of the alkyl substituent (Me, Et, (i)()Pr, Bn). The reaction of 4-chloro-N-2-phenylcyclopropyl-N-methylaniline with nitrous acid gave 4-chloro-N-methyl-N-nitrosoaniline (76%), cinnamaldehyde (55%), 3-phenyl-5-hydroxyisoxazoline (26%), and 5-(N-4-chlorophenylmethylamino)-3 phenylisoxazoline (8%). Both the selective cleavage of the cyclopropyl group from the aromatic amine nitrogen and nature of the products derived from the cyclopropane ring support a mechanism involving the formation of an amine radical cation. This step is followed by rapid cyclopropyl ring opening to produce an iminium ion with a C-centered radical which either combines with NO or is oxidized. PMID- 10813903 TI - NBS-Promoted reactions of symmetrically hindered methylphenols via p-benzoquinone methide AB - Symmetrically hindered methylphenols 1 react smoothly with NBS to form transient intermediates, p-benzoquinone methides (BM), which can be further processed to give hydroxybenzaldehydes in the presence of DMSO. This reaction is initiated by the formation of the phenoxy radical, followed by disproportionation to afford BM. None of the side-chain-brominated product is observed. The existence of BM is supported by the following observations: the formation of BM in solution can be monitored by GC and GC-MS; the electrophilic methine part participates in electrophilic aromatic substitution with anisoles to give hydroxybenzylated products 15; and the double bond character of the exocyclic methine plays a role in [4 + 2] cycloaddition with diene to afford Diels-Alder adducts. In contrast, unsymmetrically hindered or simple methylphenol (p-cresol) with NBS gives the nuclear brominated products, as usual. The energies of symmetrically hindered BMs, unsymmetrically hindered BM, and simple BM were calculated using density functional theories. Relative stabilization energies calculated at the B3LYP/6 31G//B3LYP/6-31G level by an isodesmic equation are enhanced 3-6 kcal/mol for symmetrically hindered BMs. PMID- 10813904 TI - Synthesis and characterization of monodendrons based on 9-phenylcarbazole AB - A series of 9-phenylcarbazole ethynylene monodenrons have been prepared by palladium-catalyzed coupling reactions creating well-organized arrays of redox centers. The tert-butyl groups attached to the 3,6-positions of peripheral 9 phenylcarbazole monomers provide adequate solubility to a limited degree. Trimer and 7-mer monodendrons were prepared using a monomer with 3, 3-diethyltriazene at its focal point. To facilitate purification, the synthesis of 15-mer monodendron, however, required a monomer bearing a 3-hydroxy-3-methyl-but-1-ynyl group at its focal point as a masking group for the terminal acetylene functionality. Although the solubility was limited, high generation monodendrons were found to be readily soluble in carbon disulfide, a solvent of high polarizability. Spectroscopic studies showed that there is limited through-bond conjugation over the monodendrons, but fluorescence studies suggested the presence of long-range through-space interactions in the higher members of the series. PMID- 10813905 TI - Cycloviolins A-D, anti-HIV macrocyclic peptides from Leonia cymosa. AB - Four novel anti-HIV macrocyclic peptides containing 28-31 amino acid residues, named cycloviolins A-D, have been isolated from the hitherto unstudied tropical plant Leonia cymosa. Their primary structure, including amino acid composition and sequence, was determined by a combination of MALDI-TOF and FAB MS and by enzymatic digestion of reduced derivatives, followed by Edman degradation and mass analyses. All of the cycloviolins contain six cysteines, which are present as three intramolecular disulfide bridges. Intriguingly, cycloviolins A-D showed high degrees of sequence homology to the known cyclopsychotride A and circulins A and B from the Rubiaceae family but much less homology to the varv peptides from Viola, a member of the same family (Violaceae). PMID- 10813906 TI - Access to Wieland-Miescher ketone in an enantiomerically pure form by a kinetic resolution with yeast-mediated reduction. AB - Both enantiomers of Wieland-Miescher ketone [3,4,8, 8a-tetrahydro-8a-methyl 1,6(2H,7H)-naphthalenedione], in a highly enantiomerically enriched form, became readily available by a newly developed kinetic resolution with yeast-mediated reduction. From a screening of yeast strains, Torulaspora delbrueckii IFO 10921 was selected. The collected cells of this strain, obtained by an incubation in a glucose medium, smoothly reduced only the isolated carbonyl group of the (S) enantiomer, while the (R)-enantiomer remained intact. Starting from both enantiomers ( approximately 70% ee) prepared by an established proline-mediated asymmetric Robinson annulation, the reduction with T. delbrueckii gave the (R) enantiomer (98% ee) and the corresponding alcohol (4aS,5S)-4,4a, 5,6,7,8 hexahydro-5-hydroxy-4a-methyl-2(3H)-naphthalenone (94% ee, 94% de) in preparative scale in nearly quantitative yields. An approach for the asymmetric synthesis of the Wieland-Miescher ketone was also successful. 2-Methyl-2-(3-oxobutyl)-1,3 cyclohexanedione, the prochiral precursor, was reduced with this strain to give a cyclic acetal form of (2S, 3S)-3-hydroxy-2-methyl-2-(3-oxobutyl)cyclohexanone, in a stereomerically pure form. PMID- 10813907 TI - Generalized anomeric effect in action: synthesis and evaluation of stable reducing indolizidine glycomimetics as glycosidase inhibitors. AB - A series of aminoketalic castanospermine analogues incorporating a stereoelectronically anchored axial hydroxy group at the pseudoanomeric stereocenter (C-5) have been synthesized to satisfy the need for glucosidase inhibitors that are highly selective for alpha-glucosidases. The polyhydroxylated bicyclic system was built from readily available hexofuranose derivatives through a synthetic scheme that involved (i) the construction of a five-membered cyclic (thio)carbamate or (thio)urea moiety at the nonreducing end and (ii) the intramolecular nucleophilic addition of the heterocyclic thiocarbamic nitrogen atom to the masked aldehyde group of the monosaccharide. A biological screening of the resulting reducing 2-oxa- and 2-azaindolizidines against several glycosidase enzymes is reported. PMID- 10813908 TI - A total synthesis of the methyl glycoside of ganglioside GM(1). AB - The total synthesis of the methyl glycoside of GM(1) (1b) has been accomplished. The key step in the synthesis involves the sulfonamidoglycosidation reaction, which is used to create a beta-linkage leading to a GalNAc residue joined to the C4 hydroxyl group of a galactose unit of a C3 sialylated lactosyl moiety. The "proximal hydroxyl" directing effect, which has been postulated before, manifests in this context as well leading to the preponderant formation of the beta glycoside. Together with asialo GM(1) and other substructures, the GM(1) methyl glycoside has been submitted for biological assays as potential ligands for bacterial and viral infection sites. PMID- 10813909 TI - Biosynthesis of cylindrospermopsin. AB - Studies on the biosynthesis of cylindrospermopsin (1), a potent hepatotoxin associated with the cyanobacterium Cylindrospermopsis raciborskii, indicate that 1 is an acetogenin with guanidinoacetic acid serving as the starter unit of the polyketide chain. Feeding experiments show that C14 and C15 of 1 are derived from C1 and C2 of glycine, respectively, and C4 through C13 arise from five contiguous acetate units attached head to tail. The methyl carbon on C13 originates from the C(1) pool. The starter unit, established by the incorporation of [guanidino (13)C,alpha-(15)N]-guanidinoacetic acid into N16 and C17 of 1, does not appear to be formed from glycine by known amidination pathways. The origin of the NH-CO-NH segment in the uracil ring is also unknown. PMID- 10813910 TI - Two classes of enzymes of opposite stereochemistry in an organism: one for fluorinated and another for nonfluorinated substrates. AB - Reduction of methyl ketones by dried cells of Geotrichum candidum (APG4) afforded (S)-alcohols in excellent enantiomeric excess (ee), whereas the reduction of trifluoromethyl ketones gave the corresponding alcohols of the opposite configuration also in excellent ee. The replacement of the methyl moiety with a trifluoromethyl group alters both the bulkiness and the electronic properties, the effect of which on the stereoselectivity was examined. No inversion in stereochemistry was observed in the reduction of hindered ketones such as isopropyl ketone, while the stereoselectivity was inverted in the reduction of ketones with electron-withdrawing atoms such as chlorine. The mechanism for the inversion in stereochemistry was investigated by enzymatic studies. Several enzymes with different stereoselectivities were isolated; one of them catalyzed the reduction of methyl ketones, and another with the opposite stereoselectivity catalyzed the reduction of trifluoromethyl ketones. Furthermore, both APG4 and the isolated enzyme were applied to the reduction of fluorinated ketones on a preparative scale, which resulted in the synthesis of chiral fluorinated alcohols with excellent ee. PMID- 10813911 TI - Palladium-catalyzed borylation of aryl halides or triflates with dialkoxyborane: A novel and facile synthetic route to arylboronates AB - A direct borylation of aryl halides or triflates with dialkoxyborane was investigated. The coupling reaction of pinacolborane with aryl halides or triflates in the presence of a catalytic amount of PdCl(2)(dppf) together with a base provided arylboronates in high yields. The product distributions were strongly dependent on the base employed, and the tertiary amine, especially Et(3)N, was effective for the selective formation of the boron-carbon bond. The reaction conditions were so mild that arylboronates having a variety of functional groups such as carbonyl, cyano, and nitro groups were readily prepared. PMID- 10813912 TI - Palladium-catalyzed N-arylation of sulfoximines with aryl bromides and aryl iodides AB - Various N-arylated sulfoximines have been synthesized in high yield by a direct approach which is based on a palladium-catalyzed cross-coupling strategy. Aryl bromides of variable substitution pattern were found to be the most effective coupling partners, whereas aryl iodides showed a nonpredictable behavior requiring lithium or silver salts as additives to ensure product formation in acceptable yields. Coupling of (S)-2-(2'-bromophenyl)-4-tert-butyloxazoline with enantiomerically pure (-)-(RS)-S-methyl-S-phenylsulfoximine afforded the corresponding product in good yield as a single diastereomer, showing that the palladium-catalyzed arylation proceeds in a stereospecific manner. The reaction with dibromobenzenes yielded the monosulfonimidoyl arenes in all cases, suggesting that the introduction of the sulfonimidoyl moiety deactivates the arene, thus preventing a second coupling step. PMID- 10813913 TI - Asymmetric synthesis of alpha-amino acids based on carbon radical addition to glyoxylic oxime ether AB - The first asymmetric synthesis of alpha-amino acids based on diastereoselective carbon radical addition to glyoxylic imine derivatives is reported. The addition of an isopropyl radical, generated from i-PrI, Bu(3)SnH, and Et(3)B in CH(2)Cl(2) at 25 degrees C, to achiral glyoxylic oxime ether 1 proceeded regioselectively at the imino carbon atom of the oxime ether group to give an excellent yield of the C-isopropylated product 2. The competitive reaction using glyoxylic oxime ether 1 and aldoxime ether 4 showed that the reactivity of the glyoxylic oxime ether toward nucleophilic carbon radicals was enhanced by the presence of a neighboring electron-withdrawing substituent. Thus, the alkyl radical addition to glyoxylic oxime ether 1 proceeded smoothly even at -78 degrees C, in contrast to the unactivated aldoxime ether 4. A high degree of stereocontrol in the carbon radical addition to the glyoxylic oxime ether was achieved by using Oppolzer's camphorsultam as a chiral auxiliary. The stannyl radical-mediated reaction of the camphorsultam derivative 6 with an isopropyl radical at -78 degrees C afforded a 96:4 diastereomeric mixture, 7a, of the C-isopropylated product. The reductive removal of the benzyloxy group of the major diastereomer (R)-7a, by treatment with Mo(CO)(6) and the subsequent removal of the sultam auxiliary by standard hydrolysis, afforded the enantiomerically pure D-valine (R)-12 without any loss of stereochemical purity. To evaluate the new methodology, a variety of alkyl radicals were employed in the addition reaction which gave the alkylated products 7 with excellent diastereoselectivity, allowing access to a wide range of enantiomerically pure natural and unnatural alpha-amino acids. Even in the absence of Bu(3)SnH, treatment of 6 with alkyl iodide and Et(3)B at 20 degrees C gave the C-alkylated products 7 with moderate diastereoselectivities. The use of Et(2)Zn as a radical initiator, instead of Et(3)B, was also effective for the radical reaction. The enantioselective isopropyl radical addition to 1 using (R) (+)-2, 2'-isopropylidenebis(4-phenyl-2-oxazoline) and MgBr(2) gave excellent chemical yield of the valine derivative 2 in 52% ee. PMID- 10813914 TI - Microscale determination of the absolute configuration of alpha-aryl-substituted alcohols by the CD exciton chirality method AB - The absolute configurations of a broad spectrum of aryl alcohols 1 have been determined for the first time by the CD exciton chirality method. The configurational assignment is additionally verified by computer modeling and lipase-catalyzed acetylation of the racemic alcohols. The CD-spectroscopic data have revealed that the S enantiomers of the benzoate derivatives 2 display a positive first Cotton effect and the R enantiomers a negative one at around 228 nm. Thus, the sense of the first Cotton effect of the benzoate derivative 2 allows a reliable assignment of the absolute configuration of the corresponding alcohol 1. PMID- 10813915 TI - Improved procedure for the reductive acetylation of acyclic esters and a new synthesis of ethers. AB - An optimized protocol for the DIBALH reductive acetylation of acyclic esters and diesters is described. This reductive acetylation procedure allows a wide variety of esters to be converted into the corresponding alpha-acetoxy ethers in good to excellent yields. It was found that, under mild acidic conditions, many alpha acetoxy ethers can be further reduced to the corresponding ethers. This net two step ester deoxygenation is an attractive alternative to the classical Williamson synthesis for certain ethers. PMID- 10813916 TI - Singlet-triplet energy separation of cyclobutylidene AB - Ab initio (MP2, CCSD(T)) and density functional theory (BLYP, B3LYP) calculations provide insight concerning novel aspects of structure and bonding in cyclobutylidene (1). Singlet cyclobutylidene ((1)1) adopts a bicyclobutane-like structure (C(s) symmetry) that includes a weak, transannular bonding interaction between the carbene carbon and the opposing CH(2) group. Conformational ring inversion in (1)1 occurs through a transition state of C(2)(v)() symmetry (TS(1)1) with an enthalpy barrier of approximately 3 kcal/mol. Stabilization afforded the singlet state by the transannular interaction appears to be largely offset by a loss of hyperconjugative stabilization from the adjacent C-H bonds. Triplet cyclobutylidene ((3)1) exhibits a C(2)(v)() structure and conventional bonding. The triplet state lies 5.9 kcal/mol above the singlet ground state at the CCSD(T)/TZP//CCSD(T)/DZP level of theory. The singlet-triplet energy gap of cyclobutylidene (-5.9 kcal/mol) lies between that of an acyclic analogue, dimethylcarbene (-1.6 kcal/mol), and a highly strained analogue, cyclopropylidene (-13.8 kcal/mol). The magnitude of the energy gap suggests that triplet cyclobutylidene ((3)1) will be thermally accessible under a variety of experimental conditions. PMID- 10813917 TI - New strategies for the synthesis of biologically important tetrapyrroles. The "B,C + D + A" approach to linear tetrapyrroles. AB - Linear tetrapyrroles related to phytochrome (1) were prepared in enantiospecific fashion by a new strategy beginning with ring-B,C synthons of type 19 (bis iododipyrrins). Rings A and D were elaborated by Pd(0)-mediated coupling of 19a with the appropriate alkyne acid or amide derivatives 9 and 20, followed by intramolecular cyclization (method C: BC + D + A --> ABCD). PMID- 10813918 TI - Mechanism of site-directed protein cross-linking. protein-directed selectivity in reactions of hemoglobin with aryl trimesates. AB - Site-directed cross-linking of hemoglobin has become an efficient way to produce a structurally defined altered protein with desirable functional properties. The reagent trimesoyl tris(3, 5-dibromosalicylate) (1) introduces a bis amide cross link derived from the epsilon-amino groups of the side chains of the two beta-Lys 82 residues in human hemoglobin. The basis of its specificity was investigated using a set of analogues of 1 (2-12). There are marked differences in the reaction patterns of these compounds with amino groups in hemoglobin compared to reactions with n-propylamine. The compounds that effectively modify the protein contain a carboxyl group ortho to the phenolic oxygen of the ester, while materials with meta or para carboxyl groups give little or no reaction. In contrast, the reactions with n-propylamine are slowest with the ortho carboxyl materials. Addition of the unreactive compound 5 to a solution containing hemoglobin reduces the ability of 1 to modify the protein, showing that the unreactive compound binds but does not react. On the basis of these observations and the known reaction patterns of salicylates, it is clear that the environment in the protein controls the reaction, regardless of the inherent reactivity of the reagent. We propose that the carboxyl group positions the reagent critically within the protein. Only the ortho arrangement permits transfer of the acyl function to the nucleophile. PMID- 10813919 TI - Is there a generalized reverse anomeric Effect? substituent and solvent effects on the configurational equilibria of neutral and protonated N arylglucopyranosylamines and N-aryl-5-thioglucopyranosylamines AB - The effects of substitution and solvent on the configurational equilibria of neutral and protonated N-(4-Y-substituted-phenyl) peracetylated 5 thioglucopyranosylamines (Y = OMe, H, CF(3), NO(2)) 1-4 and N-(4-Y-substituted phenyl) peracetylated glucopyranosylamines (Y = OMe, H, NO(2)) 9-11 are described. The configurational equilibria were determined by direct integration of the resonances of the individual isomers in the (1)H NMR spectra after equilibration of both alpha- and beta-isomers. The equilibrations of the neutral compounds 1-4 in CD(3)OD, CD(3)NO(2), and (CD(3))(2)CO were achieved by HgCl(2) catalysis and those of the neutral compounds 9-11 in CD(2)Cl(2) and CD(3)OD by triflic acid catalysis. The equilibrations of the protonated compounds in both the sulfur series (solvents, CD(3)OD, CD(3)NO(2), (CD(3))(2)CO, CDCl(3), and CD(2)Cl(2)) and oxygen series (solvents, CD(2)Cl(2) and CD(3)OD) were achieved with triflic acid. The substituent and solvent effects on the equilibria are discussed in terms of steric and electrostatic effects and orbital interactions associated with the endo-anomeric effect. A generalized reverse anomeric effect does not exist in neutral or protonated N-aryl-5-thioglucopyranosylamines and N arylglucopyranosylamines. The anomeric effect ranges from 0.85 kcal mol(-)(1) in 2 to 1.54 kcal mol(-)(1) in 10. The compounds 1-4 and 9-11 show an enhanced endo anomeric effect upon protonation, ranging from 1.73 kcal mol(-)(1) in 2 to 2.57 kcal mol(-)(1) in 10. We estimate the increase in the anomeric effect upon protonation of 10 to be approximately 1.0 kcal mol(-)(1). However, this effect is offset by steric effects due to the associated counterion which we estimate to be approximately 1.2 kcal mol(-)(1). The values of K(eq)(axial-equatorial) in protonated 1-4 increase in the order OMe < H < CF(3) < NO(2), in agreement with the dominance of steric effects (due to the counterion) over the endo-anomeric effect. The values of K(eq)(axial-equatorial) in protonated 9-11 show the trend OMe > H < NO(2) that is explained by the balance of the endo-anomeric effect and steric effects in the individual compounds. The trends in the values of the C(1) H(1) coupling constants for 1-4 and the corresponding deacetylated compounds 5-8 as a function of substituent and alpha- or beta-configuration are discussed in terms of the Perlin effect and the interplay of the endo- and exo-anomeric effects. PMID- 10813920 TI - The first alpha-fluoroallenylphosphonate, the synthesis of conjugated fluoroenynes, and the stereoselective synthesis of vinylfluorophosphonates using a new multifunctional fluorine-containing building block AB - Limitations on current methodologies for the introduction of CF(2) and CFH in complex alpha-fluorophosphonates led to the synthesis of a fluorine-containing building block TIPS-C&tbd1;CCFXP(O)(OEt)(2), where X = H or F. This multifunctional fluorine synthon reacts with carbonyl compounds under WHE conditions to give high yields of fluorinated conjugated enynes and enediyne. When X = F, trapping of the desilylated anion with an electrophile after TIPS removal provided exclusive access to gamma-substituted derivatives of alpha fluorophosphonates. When X = H, TBAF deprotection of the silyl group yields H(2)C=C=CFP(O)(OEt)(2) through an allenyl-propargyl resonance stabilized anion. The allene moiety has been used as template in the stereoselective synthesis of alpha-fluoro-beta, gamma-diiodopropenyl phosphonate, via electrophilic iodination, and alpha-fluoro-gamma-amino-alpha,beta-unsaturated phosphonates, including unsaturated phosphononucleosides, by nucleophilic displacement of an allylic iodide. Hydroamination of H(2)C=C=CFP(O)(OEt)(2) using secondary amines produced (Z)-alpha-fluoroenaminophosphonates, whereas Diels-Alder cycloaddition with cyclopentadiene provides the corresponding exocyclic vinylfluorophosphonate. PMID- 10813921 TI - Studies dealing with thionium ion promoted mannich cyclization reactions. AB - Treatment of several amido-substituted thioacetals with dimethyl(methylthio)sulfonium tetrafluoroborate (DMTSF) produces synthetically useful thionium ions that are intercepted by the adjacent nitrogen atom to afford both five- and six-membered alkylthio-substituted lactams as transient intermediates. Further reaction of the alkylthio-substituted lactam with DMTSF generates an N-acyliminium ion, which undergoes cyclization with the tethered aromatic ring to produce an azapolycyclic ring system. Related cyclization sequences occur when amido thioacetals possessing simple olefinic tethers were used. The overall procedure represents an efficient one-pot approach toward various nitrogen-containing ring systems. The cyclization reaction was employed for a synthesis of the core skeleton of the erythrina alkaloid family. PMID- 10813922 TI - Regiocontrolled synthetic approach to alpha,alpha'-disubstituted unsymmetrical ketones PMID- 10813923 TI - 2-(trimethylsilyl)ethanethiol in nucleoside chemistry. A short route for preparing thionucleosides and their methyl disulfides. PMID- 10813925 TI - Synthesis of 4-oxo-2-alkenylphosphonates via nitrile oxide cycloaddition: gamma acylation of allylic phosphonates PMID- 10813924 TI - Unsymmetrical diaryl ketones from arenes PMID- 10813926 TI - Asymmetric synthesis of (+)-dihydrokawain-5-ol. PMID- 10813927 TI - On the selective deprotection of trityl ethers. PMID- 10813928 TI - Oxidative activation in aromatic substitutions. Reactions Of N, N dimethylanilines with secondary anilines promoted by thallium triacetate PMID- 10813929 TI - Recent reviews. 55 AB - Reviews are listed in order of appearance in the sources indicated. In multidisciplinary review journals, only those reviews which fall within the scope of this Journal are included. Sources are listed alphabetically in three categories: regularly issued review journals and series volumes, contributed volumes, and other monographs. Titles are numbered serially, and these numbers are used for reference in the index. Major English-language sources of critical reviews are covered. Encyclopedic treatises, annual surveys such as Specialist Periodical Reports, and compilations of symposia proceedings are omitted. This installment of Recent Reviews covers principally the middle part of the 1999 Literature. Previous installment: J. Org. Chem. 1999, 64, 4(21), 7679-86. PMID- 10813930 TI - Synthesis and anion-selective complexation of cyclophane-based cyclic thioureas AB - Cyclic thiourea derivatives having three different types of cyclophane structure, ortho-meta, meta-meta, and meta-para, and a lariat-type thiourea, were synthesized, and their anion-binding ability was examined. The association constants for the complexation between the receptors and several anions in DMSO d(6) were measured by the titration method using (1)H NMR spectroscopy. All receptors, except for the meta-para cyclophane, exhibit selective binding to the dihydrogenphosphate anion, which is stronger than that of the acyclic reference compound. The lariat-type receptor binds anions even more strongly than the cyclic receptors which do not possess the third binding site. PMID- 10813931 TI - Scope and limitations of a new highly selective synthesis of unsymmetrical monomers for the synthesis of precursors toward poly(arylenevinylene)s AB - In our laboratory a precursor route to poly(p-phenylenevinylene) derivatives is developed in which unsymmetrically substituted p-xylene derivatives, possessing a benzylic sulfinylalkyl group, are used as monomers. Because of this unsymmetry, we were forced to investigate thoroughly the synthesis of these sulfoxides, as we start from symmetric and readily accessible molecules, namely, bis(halomethyl)-p xylene derivatives. In a former publication, a new extremely effective route for the production of these unsymmetrically substituted sulfinyl monomers was presented. This paper expands upon these previously reported results. To examine the scope and limitations of this elegant route, this new method was applied to the synthesis of various derivatives not included in the initial work. PMID- 10813932 TI - Ab initio analysis of pentadienyllithium, pentadienylsodium, and the pentadienyl ions AB - Ab initio calculations were used to determine the equilibrium geometries and rotational barriers of the pentadienyl cation, anion, and metalated pentadienes. Pentadienyllithium and pentadienylsodium are most stable in a U-shaped structure. This geometry is a higher energy local minimum for the pentadienyl anion and is not a stationary point for the pentadienyl cation. The atomic and group charges were analyzed by natural population analysis and were determined for each of the conformations studied. PMID- 10813933 TI - Polymer-supported BINOL ligand for the titanium-catalyzed diethylzinc addition to aldehydes: A remarkable positive influence of the support on the enantioselectivity of the catalyst AB - A new polymer-supported BINOL (1,1'-Bi-2-naphthol) was synthesized by coupling of aminomethyl polystyrene resin and (S)-2, 2'-dihydroxy-1,1'-binaphthyl-3,3' dicarboxylic acid. This new ligand was found to be more enantioselective for the asymmetric addition of diethylzinc to aldehydes than its "free" analog [Ti(BINOL)(i)PrO(2)]. A range of 57-99% ee's as well as 78-97% yields was obtained, and the electronic properties of the enantioselectivity were also observed. PMID- 10813934 TI - Photochemical alkylation of ketene dithioacetal S,S-dioxides. An example of captodative olefin functionalization AB - Radical alkylation of some ketene dithioacetal S,S-dioxides failed through the tin hydride promoted chain process but was successfully performed through stoichiometric photochemical initiation, either by electron transfer or hydrogen abstraction. In the first case, alkyl radicals were produced from tetralkylstannanes (t-Bu-, i-Pr-, n-Bu-SnR(3)) via radical cation fragmentation, while in the second case these were produced from alkanes (cyclohexane, adamantane) by benzophenone triplet. When bulky radicals (t-Bu, adamantyl) were involved, the addition occurred with complete diastereoselectivity. PMID- 10813935 TI - Preparation of hexacoordinated tris(bidentate) phosphorus compounds with 1,3,2 dioxaphosphorinane rings. NMR and X-ray crystallographic studies of their conformation AB - A series of new unstable tris(bidentate) hexacoordinated phosphorus compounds 1 with 1,3,2-dioxaphosphorinane rings was prepared and studied by NMR and X-ray crystallography. The X-ray crystal structures showed that the 1,3,2 dioxaphosphorinane ring adopts different conformations depending on the number and size of substituents at the carbon atom C14. Substituents are deployed around the phosphorus atom in a slightly deformed octahedral structure. Deviations in bond lengths around the phosphorus atom depend on the character of the bonds and the distribution of the negative charge. (1)H and (13)C NMR measurements showed that in solution the flexibility of 1,3,2-dioxaphosphorinane ring depends on the size of substituents at the carbon C14. PMID- 10813936 TI - Synthesis and conformational study of 1,3,2-oxazaphosphorino[4, 3 a]isoquinolines: a new ring system. AB - A set of 1,3,2-oxazaphosphorino[4,3-a]isoquinolines 6a,b-9a,b, a new ring system, was synthesized, and their stereochemical and conformational analyses were performed by (1)H, (13)C, and (31)P NMR methods. X-ray measurements were also carried out to confirm the stereochemical assignments and conformational results obtained by means of NMR. Intermediate coupling constants (3)J(P,H) were found for compounds 7 and 9; these do not relate to equilibria between previously reported conformers, but are indicative of new distorted conformational states in solution. The connecting isoquinoline and the steric interaction between the aromatic moiety and the Me-1 substituent can block the oxazaphosphorinane ring. The conformational behavior of compounds 6 and 8 was characterized by the usual chair-twist equilibrium. PMID- 10813937 TI - Electrostatic and electrophilic catalysis in the reductive cleavage of alkyl aryl ethers. The influence of ion pairing on the regioselectivity AB - Electrophilic and electrostatic catalysis have been identified as distinct contributions that affect the reactivity of radical anions in the reductive cleavage of alkyl aryl ethers. Two modes of mesolytic scission of these radical anions are possible: homolytic (dealkylation, a thermodynamically favored but kinetically forbidden process) and heterolytic (dealkoxylation). From our studies (alkali metal reductions, electrochemical studies, use of substrates with a preformed positive charge in certain positions of their structure) it can be concluded that the heterolytic scission is very much dependent on the electrophilic assistance by the counterion and it is only observed in contact ionic pairs with unsaturated cations (electrophilic catalysis). On the other hand, the homolytic scission is observed in solvent-separated ionic pairs, and it is especially efficient when the pair has a controlled topology with a tetralkylammonium cation (saturated cation) near the oxygen atom. The effect of the cation has, in this case, electrostatic origin (electrostatic catalysis), probably lowering the barrier of the intramolecular pi-sigma electron transfer process and thus reducing the kinetic control of the reaction in such a way that the thermodynamically more favorable process is produced. PMID- 10813938 TI - Formation of 4-methylphenanthrenes in palladium-catalyzed annulation of diethyl 2,2'-diiodo-4,4'-biphenyldicarboxylate with internal alkynes, using methyl nitrobenzoates as the methylating agent AB - The reaction of diethyl 2,2'-diiodo-4,4'-biphenyldicarboxylate (7) with diarylacetylenes in the presence of 3, 5-(NO(2))(2)C(6)H(3)CO(2)Me (MeDNB) or 4 (NO(2))C(6)H(4)CO(2)Me (MePNB), Pd(OAc)(2) (10 mol %), K(2)CO(3), and Bu(4)NBr, in DMF at 100 degrees C, gives 4-methyl-9,10-diaryl-2, 7-phenanthrenedicarboxylic acid diethyl esters in good yields. The methyl group at position 4 originates from the electron-deficient methyl nitrobenzoates. High regioselectivity for the annulation of 7 with nonsymmetrical diarylalkynes was observed with the selectivity controlled mainly by electronic factors rather than by steric factors. A competitive kinetic isotope study was carried out using a 1:1 mixture of MePNB and MePNB-d(3) in the annulation reaction of 7 which gave 4-methyl- and 4-(methyl-d(3))phenanthrene in a 67:33 ratio, leading to an apparent value of k(H)/k(D) = 1.26. Possible mechanisms for the methyl transferring process are discussed. PMID- 10813939 TI - Investigations into a biomimetic approach toward CP-225,917 and CP-263,114. AB - A biosynthesis of the structurally complex nonadride CP-225,917 (1) is outlined. A key step in this proposal is the dimerization of a C(16) anhydride derived from the condensation of lauric acid and oxaloacetic acid. An important element of this step is a templating effect imposed by two thioester linkages, reminiscent of a polyketide or fatty acid synthase pathway. On the basis of this principle, the dimerization of two C(11) anhydrides, templated by a 1,n-diol tether, leading to the core structure of CP-225,917 and CP-263,114 was investigated. PMID- 10813940 TI - Synthesis of thiamacrocycles and conformational studies on their precursors AB - 1-(4-Nitrophenyl)-7-phenylthioheptane (1) and -9-phenylthiononane (2) have been synthesized and their conformations studied in solution and in the solid state. MMX calculations suggest that the global energy minimum structures are bent in the gas phase, probably owing to edge-to-face intramolecular attractive interaction between the electron rich and the electron poor terminal aryl groups. These conformations were confirmed in solution using 2D NOESY NMR. In the solid state, 1 and 2 exist in the staggered, linear conformation, stacked head-to-tail, with the plane of the nitro group being tilted above the plane of the benzene ring. It appears that the crystal lattice forces overcome the weak edge-to-face intramolecular aromatic interactions that dominate in the gas phase and in solution. The corresponding azides were treated with trifluoromethanesulfonic acid to generate the nitrenium ions, which underwent intramolecular ring-closure to give the corresponding 17- and 19-membered ring thiamacrocycles in modest yields. These results support the suggestion that MMX calculations on appropriate model compounds may be useful in predicting which precursors will lead to macrocycles and which will not. PMID- 10813941 TI - Polymer-supported synthesis of regioregular head-to-tail-coupled Oligo(3 arylthiophene)s utilizing a traceless silyl linker AB - The solid-phase synthesis of regioregular head-to-tail-coupled oligo(3 arylthiophene)s has been achieved in high yield and purity by using a traceless silyl ether linkage. In the first step, the solution-phase synthesis of this class of conjugated oligomers was investigated. Benzyl alcohol was chosen to serve as a mimic for the anchoring group of the hydroxymethyl-substituted polystyrene matrix. The development of a novel regioselective iodination process for silyl-protected thiophenes faciliates the successful application of the solution-phase protocol to the solid phase. Satisfactory loading was obtained by reaction of chlorosilyl-functionalized 3-arylthiophene with hydroxymethyl polystyrene in the presence of imidazole. The suitability of the following iterative halogenation and Suzuki cross-coupling sequence is illustrated by the preparation of a quater(3-arylthiophene), the first regioregular head-to-tail coupled oligothiophene that is synthesized on solid support. Removal of the conjugated oligomers from the solid support could be effectively achieved by treatment with tetrabutylammonium fluoride. PMID- 10813942 TI - Polyfunctionalized cage compounds by pericyclic domino processes of 4,5 dicyanopyridazine with dienes: applications and limits AB - The title compound 1 was found to behave as an attractive masked bis-diene to give 4-oxatricyclo[4.3.1.0(3,7)]dec-8-ene, 5-aza- and 5 silatricyclo[5.3.1.0(3,8)]undec-9-ene, tricyclo[3.2.1.0(2, 7)]oct-3-ene, and tricyclo[5.3.1.0(3,8)]undec-9-ene derivatives through purely pericyclic, three step homodomino processes with diverse bis-dienophiles; whereas the reaction with myrcene (21) was characterized by a complete sitoselectivity affording compound 25, treatment of 1 with (R)-(-)-beta-citronellene (26a) gave a 3:1 mixture of the homochiral diastereomers 30a and 31a. Some limits of this methodology, mainly arising from competitive side reactions upon the key cyclohexa-1,3-diene intermediates, are emphasized. The structures of the new compounds were established on the basis of spectral data. PMID- 10813943 TI - Norbornene-constrained cyclic peptides with hairpin architecture: design, synthesis, conformation, and membrane ion transport. AB - A novel family of hairpin cyclic peptides has been designed on the basis of the use of norbornene units as the bridging ligands. The design is flexible with respect to the choice of an amino acid, the ring size, and the nature of the second bridging ligand as illustrated here with the preparation of a large number of norborneno cyclic peptides containing a variety of amino acids in ring sizes varying from 12- to 29-membered, with the choice of the second bridging ligand being a rigid norbornene (11, 13a,b), an adamantane unit (7a,b and 8), or a flexible cystine residue (4a,b and 10). The presence of built-in handles (as protected COOH groups) permits the attachment of a variety of subunits as shown here with the ligation of Leu-Leu, Val-Val, or Aib-Aib pendants in 4b, 7b, and 13b, respectively. This novel class of constrained cyclic peptides are demonstrated to adopt beta-sheet- or hairpin-like conformation as shown by (1)H NMR and CD spectra. Membrane ion-transport studies have shown that the norborneno cyclic peptides 4b and 7b containing Leu-Leu or Val-Val pendants symmetrically placed on the exterior of the ring show high efficiency and selectivity in the transport of specifically monovalent cations. This property can be attributed to the hairpin-like architecture induced by the norbornene unit since the bis adamantano peptide 15 containing two pairs of Leu-Leu pendants on the exterior is able to transport both monovalent (Na(+), K(+)) and divalent (Mg(2+)/Ca(2+)) cations. PMID- 10813944 TI - Bidirectional asymmetric allylboration. A convenient asymmetric synthesis of C(2) symmetric 3-methylenepentane-1,5-diols and rapid access to C(2)-symmetric spiroketals AB - The double allylboration of aldehydes using 1, 3-bis(diisopinocampheylboryl)-2 methylenepropanes (R,R)-3 and (S, S)-3 under Brown's salt-free conditions provides C(2)-symmetric 3-methylenepentane-1,5-diols 1 in excellent enantiomeric excess. The absolute stereochemistry of the products was confirmed by a single crystal X-ray study of bis-Mosher ester 6g. Desymmetrization and further functionalization of diol 1a were achieved by treatment of the bis-BOC carbonate 13 with IBr in toluene at -80 degrees C to give cyclic iodocarbonate 14 as a single diastereomer. This methodology is also applicable in natural product synthesis; enantiomerically pure spiroketals 1,7-dioxaspiro[5.5]undecanes 18 and 25, the latter representing an expedient synthesis of the AB ring system of the spongistatins 20, were easily accessed from simple starting materials in excellent yields and selectivities. PMID- 10813945 TI - Stereochemical analysis of deuterated alkyl chains by MS/MS AB - Vicinally deuterated sec-alkyl phenyl ethers, CH(3)(CH(2))(m)()CH(OPh)CHD(CH(2))(n)()CH(3), display significant differences in mass spectra between threo and erythro stereoisomers. MS/MS experiments, in which parent ions of a single mass are selected and their fragmentation patterns subsequently measured, show that alkene expulsion represents virtually the only decomposition pathway. Two types of MS/MS experiment are reported: mass-analyzed ion kinetic energy (MIKE) spectroscopy of metastable ions and collisionally activated decomposition (CAD) of stable ions. The expulsion of a deuterated alkene from a monodeuterated precursor yields ionized phenol, PhOH(*)()(+) (m/z 94). The expulsion of an undeuterated alkene yields PhOD(*)()(+) (m/z 95). Without exception, the ratios (PhOD(*)()(+)/PhOH(*)()(+)) from precursors in the threo series have values greater than their diastereomers in the erythro series. The ratio of ratios, r = PhOD(*)()(+)/PhOH(*)()(+) for the threo divided by PhOD(*)()(+)/PhOH(*)()(+) for the erythro, has a value of 1.2 for the 2-phenoxy-3 deuteriobutanes and larger values for all of the higher homologues up through the monodeuterated phenoxyoctanes (m + n = 4). The highest degree of stereoselectivity, r = 5.8, is measured for 3-phenoxy-4-deuteriohexane. Experiments with multiply deuterated analogues show that alkene elimination is highly regioselective, unlike the corresponding decompositions of ionized sec alcohols or their acetates. The fact that a large fraction of ionized sec-alkyl phenyl ethers undergo stereospecific syn-elimination means that mass spectrometry has a useful capacity to distinguish one isotopically labeled diastereomer from another. PMID- 10813946 TI - 1,3-Dipolar cycloadditions of diazomethane to chiral electron-deficient olefins: the origin of the pi-facial diastereoselection AB - The stereochemical outcome of diazomethane cycloadditions to several chiral electron-deficient olefins has been investigated in order to establish the origin of the pi-facial diastereoselection. Nitro olefins, vinyl sulfones, enoates, and 2-amino enoates have been used for such a purpose. These substrates have been prepared from D-glyceraldehyde acetonide through Wittig-type condensations and present an alkoxy substituent, provided by the bulky dioxolane ring, attached to the stereogenic allylic carbon. Syn-adducts have been obtained in all cases as the major isomers, independently of the Z/E stereochemistry of the double bond and the number and the nature of the substituents, the chirality of the asymmetric allylic carbon being the only thing responsible for the diastereoselection. Theoretical calculations show that steric hindrance due to the bulky dioxolane group is the main factor governing the preference for the syn attack of diazomethane to the olefinic double bond. PMID- 10813947 TI - Phenylglycine methyl ester, a useful tool for absolute configuration determination of various chiral carboxylic acids AB - A new chiral anisotropic reagent, phenylglycine methyl ester (PGME), developed for the elucidation of the absolute configuration of chiral alpha,alpha disubstituted acetic acids, has turned out to be applicable to other substituted carboxylic acids, such as chiral alpha-hydroxy-, alpha-alkoxy-, and alpha-acyloxy alpha, alpha-disubstituted acetic acids, as well as to chiral beta, beta disubstituted propionic acids. Because a carboxylic moiety is convertible from other functional groups, e.g., ozonolysis of an olefin and oxidative cleavage of a glycol, the present findings can expand the utility of the PGME method to the absolute configuration determination of various types of organic compounds, even those which initially lack oxygen functions. Several examples of the combination of chemical reactions and the PGME method are described. PMID- 10813948 TI - Modeling of protonation processes in acetohydroxamic acid AB - This work presents a theoretical study of acetohydroxamic acid and its protonation processes using ab initio methodology at the MP2(FC)/cc-pdVZ level. We find the amide form more stable than the imidic tautomer by less than 1.0 kcal mol(-)(1). For comparison with the experimental data, a three-dimensional conformational study is performed on the most stable tautomer (amide). From this study, the different barriers to rotation and inversion are determined and the intramolecular hydrogen bond between the OH group and the carbonyl oxygen is characterized. The electrostatic potential distribution shows three possible sites for electrophilic attack, but it is shown that only two of them, the carbonyl oxygen and the nitrogen atoms, are actual protonation sites. The protonation energy (proton affinity) is obtained from the results of the neutral and charged species. Proton affinities for the species charged on the carbonyl oxygen and the nitrogen atoms are estimated to be 203.4 and 194.5 kcal mol(-)(1), respectively. The development of a statistical model permits the quantification of DeltaG (gas-phase basicity) for the two protonation processes. In this way, the carbonyl oxygen protonated form is found to be more stable than that of the nitrogen atoms by 8.3 kcal mol(-)(1) at 1 atm and 298.15 K, due to the enthalpic contribution. As temperature increases, the proportion of the nitrogen protonated form increases slightly. PMID- 10813949 TI - Pairwise gibbs energies of interaction involving N-alkyl-2-pyrrolidinones and related compounds in aqueous solution obtained from kinetic medium effects AB - Kinetic solvent effects of N-alkyl-2-pyrrolidinones and structurally related compounds on the water-catalyzed hydrolysis reactions of p-methoxyphenyl dichloroacetate (MPDA), 1-benzoyl-3-phenyl-1,2, 4-triazole (BPhT), and 1-benzoyl 1,2,4-triazole (BT) in highly dilute aqueous solutions at pH 4 and 298.15 K have been determined by UV/vis spectroscopy. Using a thermodynamic description of solute-solute interactions in aqueous solutions, the kinetic results have been analyzed in terms of pairwise Gibbs energy interaction parameters: G(c) values. These are negative, indicating that hydrophobic interactions in the initial state dominate the medium effects. The interaction parameters increase in the order MPDABT>BPhT. However, when differences in reactivity and transition state effects are taken into account, it appears that BPhT is more successful in establishing hydrophobic interactions with the cosolutes than are MPDA and BT. Using the SWAG-approach for additivity of group interactions, additivity is observed for the first three consecutive CH(2) groups in the cosolute in all three hydrolysis reactions. Larger alkyl substituents cause larger retardations than anticipated on basis of this additivity. The results are explained by intramolecular destructive overlap of the polar hydration shell of the amide functionality and the apolar (hydrophobic) hydration shell of the alkyl group, which extends to the third CH(2) group in the N-alkyl group of the cosolute molecule. The inner apolar groups, therefore, have a reduced apparent hydrophobicity. More remote CH(2) groups develop independent hydrophobic hydration shells. The effect of the position of a CH(2) group in the cosolute molecule is also considered. Kinetic solvent effects with structurally related esters show that amide-amide, ester-ester, and amide-ester group interactions affect the transition state in different ways. Finally, the effects of PVP polymers on the three hydrolysis reactions have been examined. The data presented enhance the understanding of pairwise hydrophobic interactions in aqueous solutions. In addition the results provide insights into the interactions between hydrophobic and hydrophilic hydration shells as well as into the energetics of amide hydration and interactions involving amides in aqueous solution, both playing important roles in protein stabilization. PMID- 10813951 TI - Are oxocarbon dianions aromatic? AB - Assessment of the cyclic electron delocalization of the oxocarbon dianions, C(n)()O(n)()(2)(-) and their neutral counterparts C(n)()O(n)() (n = 3-6), by means of structural, energetic, and magnetic criteria, shows that C(3)O(3)(2)(-) is doubly aromatic (both sigma and pi cyclic electron delocalization), C(4)O(4)(2)(-) is moderately aromatic, but C(5)O(5)(2)(-), as well as C(6)O(6)(2)(-), are less so. Localized orbital contributions, computed by the individual gauge for localized orbitals method (IGLO), to the nucleus-independent chemical shifts (NICS) allow pi effects to be disected from the sigma single bonds and other influences. The C-C(pi) contribution to (NICS(0,pi) (i.e., at the center of the ring) in oxocarbon dianions decreases with ring size but shows little ring size effect at points 1.0 A above the ring. On the basis of the same criteria, C(4)O(4) exhibits cyclic electron delocalization due to partial occupancy of the sigma CC bonds. However, the dissociation energies of all the neutral oxocarbons, C(n)()O(n)(), are highly exothermic. PMID- 10813950 TI - cis,cis- and trans,trans-ceratospongamide, new bioactive cyclic heptapeptides from the Indonesian red alga Ceratodictyon spongiosum and symbiotic sponge Sigmadocia symbiotica. AB - Chemical investigation of the marine red alga (Rhodophyta) Ceratodictyon spongiosum containing the symbiotic sponge Sigmadocia symbiotica collected from Biaro Island, Indonesia, yielded two isomers of a new and bioactive thiazole containing cyclic heptapeptide, cis,cis-ceratospongamide (1) and trans, trans ceratospongamide (2). Isolation of these peptides was assisted by bioassay-guided fractionation using a brine shrimp toxicity assay (Artemia salina). The structures of the ceratospongamides, which each consist of two L-phenylalanine residues, one (L-isoleucine)-L-methyloxazoline residue, one L-proline residue, and one (L-proline)thiazole residue, were established through extensive NMR spectroscopy, including (1)H-(13)C HMQC-TOCSY, and (1)H-(15)N HMBC experiments, as well as chemical degradation and chiral analysis. cis,cis- and trans,trans ceratospongamide are stable conformational isomers of the two proline amide bonds. Molecular modeling of these two ceratospongamide isomers showed the trans, trans isomer to be quite planar, whereas the cis,cis isomer has a more puckered overall conformation. trans,trans-Ceratospongamide exhibits potent inhibition of sPLA(2) expression in a cell-based model for antiinflammation (ED(50) 32 nM), whereas the cis,cis isomer is inactive. trans,trans-Ceratospongamide was also shown to inhibit the expression of a human-sPLA(2) promoter-based reporter by 90%. PMID- 10813952 TI - Practical asymmetric synthesis of (S)-MA20565, a wide-spectrum agricultural fungicide. AB - A practical asymmetric synthesis of a wide-spectrum agricultural fungicide, (S) MA20565 (1), is described. The convergent synthesis was achieved starting from commercially available 3-(trifluoromethyl)aniline (7) in 44% overall yield through five steps and 2-bromobenzaldehyde (9) in 48% overall yield through four steps, respectively. (S)-O-[1-(3-Trifluoromethylphenyl)ethyl]hydroxylamine (2), a key intermediate of 1, was prepared via ruthenium(II)-catalyzed asymmetric transfer hydrogenation of 1-(3-trifluoromethylphenyl)ethanone (6) followed by chlorination using methanesulfonyl chloride and oxyamination using potassium acetohydroxamate with high level of stereocontrol. PMID- 10813953 TI - Absolute configuration of aflastatin A, a specific inhibitor of aflatoxin production by Aspergillus parasiticus. AB - Aflastatin A (1) is a specific inhibitor of aflatoxin production by Aspergillus parasiticus. It has the novel structure of a tetramic acid derivative with a long alkyl side chain. The absolute configurations of 29 chiral centers contained in 1 were chemically elucidated in this study. First, four small fragment molecules were prepared from 1 or its methyl ether (2), and their absolute structures were assigned as N-methyl-D-alanine, (2S,4R)-2, 4-dimethyl-1,6-hexanediol dibenzoate, (R)-3-hydroxydodecanoic acid, and (R)-1,2,4-butanetriol tribenzoate. Next, an acyclic fragment molecule 3 with 13 chiral centers was obtained from 1 by NaIO(4) oxidation, and its relative stereochemistry was elucidated by J-based configuration analysis. By analyzing coupling constants of (3)J(H,H) and (2,3)J(C,H) and ROE data, the relative configuration of 3 was verified. Finally, by further J-based configuration analysis using a fragment molecule 7 prepared from 2 with 28 chiral carbons, all relative configurations in the alkyl side chain of 1 were clarified. By connecting these relative configurations with the absolute configurations of first four fragment molecules, the absolute stereochemistry of 1 was fully determined. PMID- 10813955 TI - Photochemical dimerization of acephenanthrylene and the heavy atom effect AB - The solution-phase photodimerization of acephenanthrylene (1) in cyclohexane, chloroform, dibromomethane, and ethyl iodide with visible light (lambda = 419 nm) occurs to form cis (2) and trans (3) photodimers in ratios that vary with the solvent and with the presence or absence of oxygen. The photodimerization exhibits a significant heavy atom effect and also is sensitive to selective quenching by dissolved oxygen, in which only cis stereoisomers are produced. Irradiation of the dimers (lambda = 254 nm) shows that the trans dimers cleave more rapidly than the cis dimers. This is attributed to stabilization of the excited state in the cis dimers. PMID- 10813954 TI - Peloruside A: a potent cytotoxic macrolide isolated from the new zealand marine sponge Mycale sp. AB - A novel, polyoxygenated, pyranose ring containing 16-membered macrolide peloruside A (1) exhibiting cytotoxic activity in the nanomolar range was isolated from the New Zealand marine sponge Mycale sp. The structure of 1 and relative stereochemistry of the 10 stereogenic centers were determined on a 3 mg sample using a variety of spectroscopic methods. Compound 1 was isolated along with the previously reported cytotoxins mycalamide A (2) and pateamine (3) from a single specimen of this sponge. PMID- 10813956 TI - Diels-alder reactions with 2-(Arylsulfinyl)-1,4-benzoquinones: effect of aryl substitution on reactivity, chemoselectivity, and pi-facial diastereoselectivity AB - Diels-Alder reactions of (SS)-2-(2'-methoxynaphthylsulfinyl)-1, 4-benzoquinone (1b), 2-(p-methoxyphenylsulfinyl)-1,4-benzoquinone (1c), and 2-(p nitrophenylsulfinyl)-1,4-benzoquinone (1d) with cyclopentadiene are reported. These cycloadditions allowed the highly chemo- and stereoselective formation of both diastereoisomeric endo-adducts resulting from reaction on the unsubstituted double bond C(5)-C(6) of quinones working under thermal and Eu(fod)(3)- or BF(3).OEt(2)-catalyzed conditions. The synthesis of endo-adduct [4aS,5S,8R,8aR,SS]-9d resulting from cycloaddition on the substituted C(2)-C(3) double bond was achieved in a chemo- and diastereoselective way from quinone 1d in the presence of ZnBr(2). The reactivity and selectivity of the process proved to be dependent on the electron density of the arylsulfinyl group. PMID- 10813957 TI - Chloptosin, an apoptosis-inducing dimeric cyclohexapeptide produced by Streptomyces. AB - In the course of screening for apoptosis-inducing agents, chloptosin (1) was isolated from the culture broth of Streptomyces. The dumbbell-type structure of the dimeric cyclohexapeptide consisting of D-valine, (3S)- and (3R)-piperazic acids, O-methyl-L-serine, D-threonine, and (2S,3aR,8aR)-6-chloro-3a-hydroxy 2,3,3a, 8a-hexahydropyrrolo[2,3-b]indole-2-carboxylic acid was elucidated by spectroscopic and chemical degradation studies. The amino acid components in each cyclohexapeptide domain were presented in alternating R and S configurations. Chloptosin (1) was found to induce apoptotic activity in apoptosis-resistant human pancreatic adenocarcinoma cell line AsPC-1 and showed a strong antimicrobial activity against Gram-positive bacteria including methicillin resistant Staphylococcus aureus. PMID- 10813958 TI - Oxidative homocoupling of chiral 3-arylpropanoic acid derivatives. Application To asymmetric synthesis of lignans AB - The oxidative homocouplings of lithium enolates of (4S)-3-(3-arylpropanoyl)-4 isopropyl-2-oxazolidinones and (4R, 5S)-1-(3-arylpropanoyl)-3,4-dimethyl-5-phenyl 2-imidazolidinones gave the corresponding R,R-dimers stereoselectively with TiCl(4), PhI(OAc)(2), or CuCl(2) as an oxidant. The stereoselectivity can be explained by a radical coupling mechanism. Optically active dibenzylbutyrolactone lignans, such as (-)-hinokinin and (-)-dimethylmatairesinol, and dibenzylbutanediol lignans, such as (-)-dihydrocubebin and (-) dimethylsecoisolariciresinol, were synthesized from the major R,R-dimers. The oxidative coupling of (4R, 5S)-1-(3-arylpropanoyl)-3,4-dimethyl-5-phenyl-2 imidazolidinones with LDA-I(2) gave R,S-dimers mainly, and this result can be explained by an S(N)2 mechanism. PMID- 10813959 TI - Stereoselective reaction of alpha-sulfinyl carbanion derived from chiral 2 (Trialkylsilyl)ethyl sulfoxides: evidence for a novel silicon-oxygen interaction AB - Reactions of alpha-sulfinyl carbanions, derived from p-tolyl sulfoxides bearing various alkyl groups, with various electrophiles were examined. The reaction of alpha-sulfinyl carbanions, derived from the beta-silylethyl sulfoxides, with ketones or trimethyl phosphate, gave the syn products with high stereoselectivity. Interaction between the silicon in the trialkylsilyl group and the carbonyl oxygen in nucleophiles was postulated to stabilize the transition state, leading preferably to the syn diastereisomers. This novel silicon-oxygen interaction was supported by an MO calculation study using the MOPAC 93/PM3 and the Gaussian 94 Beche3LYP/3-21+G methods. PMID- 10813960 TI - Three-component coupling of acylphosphonates and two carbonyl compounds promoted by low-valent samariums: one-Pot synthesis of beta-hydroxyphosphonates AB - Three-component coupling of acylphosphonates and two carbonyl compounds leading to beta-hydroxyphosphonates has been achieved with low-valent samariums. Thus, acylphosphonates reacted with aldehydes in the presence of semicatalytic amounts of samarium metal or SmI(2) to give acyloxyphosphonates in good yields. The second coupling reaction of the acyloxyphosphonates with aldehydes or ketones promoted by SmI(2) afforded beta-hydroxyphosphonates instead of olefins. Moreover, these two reactions could be carried out in one pot. PMID- 10813961 TI - Stereoselective preparation of enantiomerically pure annulated carbohydrates using ring-closing metathesis AB - Ring-closing metathesis has been applied to a series of glucose derivatives to produce cyclopentene derivatives 5a and 5b, cyclohexene derivatives 8 and 9, cycloheptene 12, and cyclooctene 14. Spirocyclic dihydrofurans 19, 26a, and 26b, along with dihydropyran 22, were also produced. A range of fused oxepine derivatives 29a-c and one oxo-cyclononene 31 were also prepared. Cyclopentene 5b was subjected to a sequence of hydrogenation, NBS bromination, and treatment with powdered zinc to furnish the ring-expanded product 35. No such ring expansion occurred when the cyclohexaannulated compound 8 was treated with NBS followed by powdered zinc, leading to aldehyde 39. The spiro dihydrofuran derivative 19 was converted to the aldehyde 42 via the same reaction sequence used to fragment cyclopentene derivative 5b. PMID- 10813962 TI - Regio- and diastereocontrol in carbonyl allylation by 1-halobut-2-enes with Tin(II) halides AB - Regio- and diastereoselective carbonyl allylations of 1-halobut-2-enes with tin(II) halides are described. Tin(II) bromide in a dichloromethane-water biphasic system is an effective reagent for unusual alpha-regioselective carbonyl allylation of 1-bromobut-2-ene to produce 1-substituted pent-3-en-1-ols. The addition of tetrabutylammonium bromide (TBABr) to the biphasic system produces 1 substituted 2-methylbut-3-en-1-ols via usual gamma-addition which is opposite to the alpha-addition without TBABr. The gamma-addition to aromatic aldehydes exhibits anti-diastereoselectivity, while that to aliphatic aldehydes is not diastereoselective. The allylation of benzaldehyde by 1-chlorobut-2-ene in 1,3 dimethylimidazolidin-2-one (DMI) does not occur with tin(II) chloride or bromide but does proceed with tin(II) iodide and exhibits gamma-syn selectivity which is unusual for a Barbier-type carbonyl allylation. In the carbonyl allylation by 1 chlorobut-2-ene with any tin(II) halide, the addition of tetrabutylammonium iodide (TBAI) accelerates the reaction and enhances gamma-syn selectivity. The use of tin(II) iodide and TBAI produces 2-methyl-1-phenylbut-3-en-1-ol with high yield and high syn-diastereoselectivity. The syn-diastereoselective carbonyl allylation of 1-chlorobut-2-ene using tin(II) iodide, a catalytic amount of TBAI, and NaI in DMI-H(2)O is applied to various aldehydes. PMID- 10813963 TI - Sequential 1,3-dipolar cycloadditions in the synthesis of bis-isoxazolo substituted piperidinones AB - A strategy for the efficient synthesis of novel bis-isoxazolo substituted piperidinones has been developed. The protocol consists of the Michael addition of an unsaturated alkoxide to beta-nitrostyrene followed by an intramolecular nitrile oxide cycloaddition (INOC) or an intramolecular silyl nitronate olefin cycloaddition (ISOC) to give III. Grignard addition to this isoxazoline intermediate followed by DCC coupling of the resulting isoxazolidine with nitroacetic acid gave II, and a second intramolecular cycloaddition via 1,3 dipoles result in the formation of the targeted novel tetracycles (I). A solid supported scavenger was employed to increase the efficiency and yield of the Michael addition step. PMID- 10813964 TI - Asymmetric synthesis of 4-deoxyverrucarol via two types of ring expansion reactions AB - Asymmetric synthesis of a trichothecane analogue, 4-deoxyverrucarol (2), was carried out through two types of ring expansion reactions. First, synthesis of the racemate of 2 was investigated. Thus, 1-[1-(tert-butyldimethylsiloxy)-ethyl] 1-methoxycarbonyl-2-hexen-4-on e (10), prepared by Diels-Alder reaction, was converted into the cyclopropylidene 15. The cyclobutanone (+/-)-18 was obtained from 15 via dihydroxylation, followed by successive treatments with SO(2)Cl(2) in the presence of imidazole and Florisil. After transformation of (+/-)-18 into the vinylcyclobutanol (+/-)-19, the second ring expansion reaction was performed with Pd(OAc)(2) to provide the cyclopentanone (+/-)-20. The product was converted into the racemate of 4-deoxyverrucarol (2) through the cyclohexenone (+/-)-22, but the diastereoselectivity during the introduction of the double bond was unsatisfactory. The selectivity was improved in the case of the asymmetric synthesis. The optically active cyclobutanone (+)-18 was prepared via AD reaction of 15 with 73% ee. After the transformation of (+)-18 into the cyclohexanone (-) 30 through the palladium-mediated ring expansion reaction, (-)-30 was subjected to the diastereoselective deprotonation reaction using the chiral amide. The key synthetic intermediate (-)-25 of 4-deoxyverrucarol (2) was synthesized in an optically pure form by taking advantage of a kind of kinetic resolution that occurred during the deprotonation step. PMID- 10813965 TI - Restricted motion of guests confined in carceplexes and capsules. AB - Guest orientation within carceplexes and capsules was determined qualitatively from NMR data, and the molecular mobility of guests was determined via coalescence of (1)H NMR signals. Both are highly dependent on guest size and shape, as is interconversion of twistomers. Incarceration of 1,4-thioxane results in a large (1.8 kcal/mol) constraint on thioxane's conformational mobility (chair to-chair interconversion). Similar conformational constraints (1.6 kcal/mol) were determined for 1,4-dioxane both when incarcerated in carceplex 1b and when encapsulated reversibly in capsule 3b. Encapsulation-induced conformational constraints of this magnitude are unprecedented, and are particularly striking for the noncovalently linked capsules. PMID- 10813967 TI - Inter- and intramolecular pathways for the formation of tetrahydrofurans from beta-(Phosphatoxy)alkyl radicals. Evidence for a dissociative mechanism AB - beta-(Phosphatoxy)alkyl radicals generated by photolysis of Barton PTOC esters in the presence of allyl alcohol and tert-butyl mercaptan undergo nucleophilic substitution followed by 5-exo-trig radical ring closure leading to tetrahydrofurans in good yield and with high trans selectivity. beta (Phosphatoxy)alkyl radicals obtained by intramolecular hydrogen 1,5-abstraction with an alkoxyl radical undergo nucleophilic displacement providing tetrahydrofurans. The ensemble of results, including the effects of leaving groups and substituents, strongly support a dissociative mechanism for these radical nucleophilic displacement reactions. PMID- 10813966 TI - Practical synthesis of (R)-4-mercaptopyrrolidine-2-thione from L-aspartic acid. Preparation of a novel orally active 1-beta-methylcarbapenem, TA-949. AB - A facile and economical synthesis of a novel orally active 1-beta methylcarbapenem, TA-949 (1), is described. The key process involves an efficient synthesis of the C-2 side chain (R)-4-mercaptopyrrolidine-2-thione 2 from L aspartic acid and the construction of the 1-beta-methylcarbapenem skeleton. The mercapto group of 2 with an R-configuration was formed via deaminative bromination of the amino group of L-aspartic acid beta-methyl ester hydrochloride 12 followed by a complete S(N)2-type substitution with potassium benzenemethanethiolate. High-yield amination and cyclization of the chloride 15 to the pyrrolidin-2-one 16 was accomplished by a simple treatment with ammonia. Thiation of 16 and the Birch reduction of the resultant thiolactam 18 provided the C-2 side chain 2 in high yield with the asymmetric center retained as such. The side chain 2 was installed into the 1-beta-methylcarbapenem skeleton either by coupling with the vinyl phosphate 5 or by the use of the counterattack strategy involving the Dieckmann-type cyclization of the thioester 8. Removal of the protective groups of the coupling product 6 followed by esterification provided TA-949 (1) in high yield. PMID- 10813968 TI - Improved synthesis of isogranulatimide, a G2 checkpoint inhibitor. Syntheses Of didemnimide C, isodidemnimide A, neodidemnimide A, 17-methylgranulatimide, and isogranulatimides A-C. AB - A concise, improved synthesis of isogranulatimide (6), a naturally occurring substance with G2 checkpoint inhibition activity, is described. Also reported are the syntheses of didemnimide C (18), isodidemnimide A (24), neodidemnimide A (36), 17-methylgranulatimide (9), and isogranulatimides A (10), B (11), and C (12). Compounds 9-12, congeners of isogranulatimide (6), are now available for biological evaluation. PMID- 10813969 TI - Relative binding affinities of alkali metal cations to AB - The binding affinity and selectivity of a new ionophore, [1(8)]starand (1), toward alkali metal cations in methanol were examined through NMR titration experiments and free energy perturbation (FEP) and molecular dynamics simulations. The preference was determined to be K(+) > Rb(+) > Cs(+) > Na(+) >> Li(+) in both FEP simulations and NMR experiments. The FEP simulation results were able to predict the relative binding free energies with errors less than 0.13 kcal/mol, except for the case between Li(+) and Na(+). The cation selectivity was rationalized by analyzing the radial distribution functions of the M-O and M-C distances of free metal cations in methanol and those of metal ionophore complexes in methanol. PMID- 10813971 TI - Stereoelectronic effects in negatively and positively (Protonated) charged species. Ab initio studies of the gauche effect in 1,4-dioxa systems AB - A computational study ab initio of the conformational dependence of proton affinities of 2-methoxyethoxide (MEO), dimethoxyethane (DME), and 1,4-dioxane has been carried out at the MP2/6-31+G level of theory. The results were discussed in comparison with reference systems, from simple alkoxides and ethers to anomeric moieties, in open, cyclic, and bicyclic molecules. The COCCOC species are stronger bases than the COCOC (anomeric) ones and approach regular ethers in their strength. The gauche forms in MEO and DME are altogether stronger bases than the anti forms, and anti (equatorial) protonation is preferred over gauche (axial), unless ditopic protonation is possible, like in aga-DME or cis tetraoxadecalin. The gauche effect plays a significant role in the formation, relative stability, and reactivity of the charged species. PMID- 10813970 TI - Wittig reactions on photoprotoporphyrin IX: new synthetic models for the special pair of the photosynthetic reaction center. AB - A first example of spirochlorin-chlorin dimer with fixed distances and orientations as potential model for the "special pair" of the photosynthetic reaction center is discussed. For the preparation of such a novel structure, the Wittig reagent of the desired "spacer" 5 was reacted with photoprotoporphyrin IX dimethyl ester 3 to produce the intermediate dimer 6, which on intramolecular [4 + 2] Diels-Alder cycloaddition gave an unexpected spirochlorin-chlorin dimer 9. Dehydration of dimer 6 under acid-catalyzed conditions generated the corresponding spirochlorin-porphyrin dimer 16 in quantitative yield. The asymmetry in dimer 6 caused by the biphenyl-type anisotropic effect was confirmed by NMR and model studies. The formation of dihydrobenzoporphyrin 14 by reacting chlorin 3 with the phosphonium salt of p-methylbenzylbromide 10 and isolation of 8-phenanthrenevinylporphyrin 19 from chlorin 7 further confirmed our proposed mechanism for the formation of a spirochlorin-chlorin dimer 9. Following a similar approach, chlorin 3 on reacting with bis-phosphonium salt of 4, 4' bischloromethylbiphenyl produced conjugated chlorin dimer 25. The spectroscopic data obtained from these dimers suggest that, in these compounds, the individual chromophores are not behaving as an individual molecule, but as a single macrocycle. To examine whether the pi-pi interaction exhibited by dimer 9 resembles the structural arrangement of bacteriochlorophylls in reaction center (RC), we investigated the geometrical parameters used to characterize the pi-pi interactions in tetrapyrrolic macrocycles. Starting from the crystallographic coordinates of 9, the molecular mechanics energy minimization was performed to obtain the model dimer structure. The geometrical parameters that measure the single pyrrole ring overlap were used to compare the model structure with the crystallographic coordinates of the special pair in photosynthetic reaction center. The results indicated that the ring A of spirochlorin and the ring C of chlorin in our model dimer 9 mimic the ring A-ring A interaction found in the crystallographic special pairs, which are strategically placed by the surrounding photosynthetic reaction center protein matrix. PMID- 10813972 TI - Exceptionally pyramidalized olefins: A theoretical study of the cyclopropenyl fused tricycles Tricyclo AB - RHF, MP2, and TCSCF ab initio theory and B3LYP, B3PW91, and SVWN density functional theory were used to study the series of cyclopropenyl-fused tricycles 9-12. In each of 9-12, the cyclopropenyl double bond is exceptionally pyramidalized (butterfly angle psi approximately 41-50 degrees ) with both endo and exo bent isomers. In the norbornyl systems (9 and 10), the endo bent isomers are more stable than the exo bent isomers, whereas in the bicyclo[2. 2.2]octadiene 12 the reverse is true with the exo bent isomer being the low energy form. The activation barriers for the endo/exo interconversions are calculated to be relatively low (DeltaH() approximately 6-13 kcal/mol). PMID- 10813973 TI - Effective synthesis of aryl ethers and coumaranones employing the palladium catalyzed enyne-diyne AB - Alkyl-, alkoxy-, and aryloxy-substituted conjugated enynes 1 in the presence of Pd(PPh(3))(4) catalyst smoothly underwent the regiospecific [4 + 2] cycloaddition reaction with conjugated alkyl- and alkoxy-substituted symmetric diynes 2 to give multisubstituted aryl ethers 3 in good to high yields. Benzannulation of enynes 1d-g with unsymmetric diyne 6, possessing alkyl and alkoxy groups at acetylenic terminii, in most cases produced an aromatic product 8 with an alkoxy group of diyne attached to the ethynyl moiety of the aromatic product. Remarkably, alkoxy substituted diynes 2c and 6 underwent the benzannulation reaction with 1 at an unusually low temperature of 0 degrees C! One-pot consecutive benzannulation of alkyl-substituted enynes 1d,e and alkoxy-substituted enynes 1f,g with alkoxy substituted diynes 2c and 6 followed by protonolysis with TsOH afforded coumaranones 9a-c and 10 in reasonable to high overall yields. PMID- 10813974 TI - Kinetics of the base-catalyzed permanganate oxidation of benzaldehyde AB - The kinetics of the base-catalyzed permanganate oxidation of benzaldehyde have been reexamined. The rate is proportional to the first power of the aldehyde and permanganate concentrations, and there are terms that are zero order, first order, and second order in hydroxide ion. The reaction has an isotope effect, and the effect of substituents gives rho = +1.58. The possible mechanisms for the reaction are discussed in the context of ab initio calculations at the B3P86/6 311+G and MP2/6-31G theoretical levels, and both one- and two-electron processes are possible. Benzaldehyde hydrate dianion is calculated to have a remarkably small C-H bond dissociation energy. PMID- 10813975 TI - The origin of the alpha-effect: dissection of ground-state and transition-state contributions AB - The origin of the alpha-effect has been probed through a combination of calorimetric and kinetic studies involving butane-2,3-dione monoximate as alpha nucleophile and p-chlorophenoxide as normal nucleophile in the reaction with p nitrophenyl acetate in DMSO-H(2)O mixtures, which has been shown to exhibit a bell-shaped profile in the alpha-effect with solvent composition. The study, involving determination of enthalpies of solution and activation parameters, has allowed a dissection of contributions to the alpha-effect of ground-state destabilization and transition-state stabilization in these DMSO-H(2)O solvent media. It has been found that over the solvent composition 0-50 mol % DMSO desolvation of the alpha-nucleophile is the main driving factor to the increasing alpha-effect. However, in solvent mixtures covering 50-90 mol % DMSO the thermodynamic activation parameters suggest an interplay of factors that result in the bell-shaped alpha-effect profile. Discussion is presented that includes possible medium-dependent nonsynchronicity of nucleophile desolvation and bond formation for the alpha-nucleophile. PMID- 10813976 TI - A convenient method for synthesizing 2-aryl-3-hydroxy-4-oxo-4H-1-benzopyrans or flavonols. PMID- 10813977 TI - An efficient preparation of 2-C-methyl-D-erythritol 4-phosphoric acid and its derivatives PMID- 10813978 TI - Synthesis of viologens with extended pi-conjugation and their photochromic behavior on near-IR absorption PMID- 10813979 TI - Synthesis of 5- and 6-hydroxymethylfluorescein phosphoramidites. PMID- 10813980 TI - Practical synthesis of selectively functionalized cavitands PMID- 10813981 TI - Simple and efficient generation of alpha-fluoromalonaldehyde from fluorinated enol sulfonate and its reaction with acyl chlorides leading to (Z)-beta-acyloxy alpha-fluoroacrylaldehydes PMID- 10813982 TI - A practical synthesis of (Z)-debromohymenialdisine. PMID- 10813984 TI - In situ manganese dioxide alcohol oxidation-wittig reactions: preparation of bifunctional dienyl building blocks PMID- 10813983 TI - An efficient synthesis of antibiotic (-)-A26771B. PMID- 10813985 TI - Samarium diiodide-promoted cyclization of N-(omega-Iodoalkyl)imides to polyhydroxylated indolizidinones and pyrrolizidinones: synthesis of (+) lentiginosine PMID- 10813986 TI - Photolysis of bis(organosilyl)imines PMID- 10813987 TI - Highly stereoselective addition-elimination reaction of nucleophiles with ethyl 3,3-difluoro-2- PMID- 10813988 TI - Stereoselective synthesis of stereotriad subunits of polyketides through additions of nonracemic allenylsilanes to (R)- and (S)-2-methyl-3-oxygenated propanals PMID- 10813989 TI - A general and facile synthesis of substituted furans by palladium-catalyzed cycloisomerization of (Z)-2-En-4-yn-1-ols PMID- 10813990 TI - Synthesis of mimosamycin. AB - Mimosamycin (1) was synthesized in eight steps with an overall yield of 13% from 2-methoxy-3-methyl-1,4-benzoquinone by regioselective introduction of a chloromethyl group at C-6 and a methoxycarbonylmethyl group at C-5 and subsequent reaction of the intermediate methyl (o-(chloromethyl)phenyl)acetate derivative 16 with methylamine. Oxidation of the 5,7,8-trimethoxy-2,6-dimethyl-1, 4 dihydroisoquinoline-3(2H)-one 17 thus obtained, using cerium(IV) ammonium nitrate as a selective oxidizing agent, gave mimosamycin (1) in good overall yield. PMID- 10813991 TI - Synthesis of two naphthoquinone antibiotics, dehydroherbarin and 6 deoxybostrycoidin. AB - The synthesis of two naphthoquinone antibiotics, dehydroherbarin (7) and 6 deoxybostrycoidin (5), was accomplished by reaction of 3-acetonyl-2-bromomethyl 6,8-dimethoxy-1,4-naphthoquinone (23) with either triethylamine or ammonia, respectively. This is the first report on their synthesis. PMID- 10813994 TI - Cyclizations of substituted benzylidene-3-alkenylamines: synthesis of the tricyclic core of the martinellines. AB - The martinellines (1 and 2) are natural products that possess both interesting biological activity and chemical structure. During the investigation of a hetero Diels-Alder route to these molecules, alternate Lewis acid-dependent cyclizations of (2'-amino-N'-tert-butoxycarbonyl-5'-chlorobenzylidene)-3-butenylamine (10) were observed. The reaction of a variety of imines with TMSOTf or TiCl(4) led to the formation of different heterocycles including iminodibenzo[b,f][1,5]diazocines, hexahydropyrido[1, 2-c]quinazolin-6-ones, tetrahydropyrrolo[1,2-c]quinazolin-5-ones, 2-arylpiperidines, and 2 arylpyrrolidines. Tetrahydropyrrolo[1, 2-c]quinazolin-5-one 54, obtained via this new methodology, was used as an intermediate in the synthesis of the tricyclic ring system (65) of the martinellines. PMID- 10813998 TI - Enantiomeric syntheses of conformationally restricted D- and L-2', 3'-dideoxy 2',3'-endo-methylene nucleosides from carbohydrate chiral templates. AB - D- and L-2',3'-dideoxy-2',3'-endo-methylene nucleosides were synthesized as potential antiviral agents. The key intermediates 5-O-tert-butyldiphenylsilyl-D- and L-2,3-dideoxy-2, 3-endo-methylenepentofuranoses (20 and 33, respectively) were obtained by selective protection of the D- and L-2,3-dideoxy-2, 3-endo methylenepentose derivatives 19 and 32 which were prepared from 1,2:5,6-di-O isopropylidene-D-mannitol and L-gulonic gamma-lactone, respectively, and converted to 5-O-tert-butyldiphenylsilyl-D- and L-2,3-dideoxy-2, 3-endo methylenepentofuranosyl acetates (21 and 34, respectively) or the chlorides 22 and 35. The acetates and chlorides were condensed with pyrimidine and purine bases by Vorbruggen conditions or S(N)2-type condensation. Vorbruggen conditions using the acetates gave mostly alpha-isomers. In contrast, S(N)2-type condensation using the chlorides greatly improved the beta/alpha ratio. From the synthesis, several D- and L-2',3'-dideoxy-2',3'-endo-methylene nucleoside analogues have been obtained, and their structures have been elucidated by NMR spectroscopy and X-ray crystallography. The synthesized D- and L-adenine derivatives were tested as substrates of adenosine deaminase, which indicated that the D-adenosine derivative 4a was a good substrate of a mammalian adenosine deaminase from calf intestinal mucosa (EC 3.5.4.4) while its L-enantiomer 10a was a poor substrate. Either the D-adenine derivative 4a or its L-enantiomer 10a did not serve as an inhibitor of the enzyme. PMID- 10814001 TI - Synthesis of 10,11-dihydroleukotriene B(4) metabolites via a nickel-catalyzed coupling reaction of cis-bromides and trans-alkenyl borates. AB - Synthesis of 10,11-dihydro-, 10,11,14,15-tetrahydro-, and 10, 11-dihydro-12 oxoleukotriene B(4) compounds (2, 4, 5) was accomplished stereoselectively by using the nickel-catalyzed coupling reaction illustrated in Scheme 1. The C(1) C(7) fragments, TBS ether 10a for 2 and 4 and ethyoxyethyl (EE) ether 10b for 5, were prepared in enantiomerically pure forms (>99% ee) by a modified literature procedure (ref 11a). On the other hand, boronate esters 11a and 11b, which correspond to the C(8)-C(20) parts of 2 and 4, respectively, were synthesized from (R)-epichlorohydrin (18) of 99% ee. Briefly, 18 was converted into acetylenes 24 and 32 through epoxide ring-opening with LiC triple bond CC(5)H(11)/BF(3).OEt(2) or C(7)H(15)MgBr/CuCN. Hydroboration of these acetylenes with (+)-(Ipc)(2)BH followed by reaction with MeCHO afforded the corresponding diethyl boronates, which upon ligand exchange with Me(2)C(CH(2)OH)(2) furnished boronate esters 11a and 11b in 75% and 77% yields, respectively. In a similar manner, racemic boronate ester rac-11a, an intermediate for synthesis of 5, was prepared from racemic epichlorohydrin. For synthesis of 2, borate 25 was generated from 11a (1.5 equiv) and MeLi (1.6 equiv). Without isolation, 25 was submitted to reaction with 10a (1 equiv) in the presence of a Ni(0) species at room temperature overnight to afford 26, which upon treatment with TBAF furnished 2 in 64% yield from 10a. Similarly, 11b and 10a furnished 4 in good yield. To synthesize 5, rac-11a and EE ether 10b were joined by the coupling reaction to produce 39, which was transformed into 40 by desilylation with TBAF. After hydrolysis of 40, oxidation with PDC followed by deprotection of the EE group furnished 5 in 36% yield from 40. In addition, 2 was converted into amide 3 in 92% yield. PMID- 10814002 TI - New stereoselective route to the epoxyquinol core of manumycin-type natural products. Synthesis of enantiopure (+)-bromoxone, (-)-LL-C10037 alpha, and (+)-KT 8110. AB - A practical route is decribed for the preparation of the C(7)N core of manumycin type compounds. Starting from p-benzoquinone, optically pure compounds in both forms can be prepared via enzymatic resolution of a derived diacetoxy conduritol. A diepoxy aminoinositol is accessible which can function for formation of enantiopure epoxyquinones and quinols. Examples are given for acylation reactions of this amine with several acyl derivatives. With this approach (-)-LL C10037alpha and quinones such as (+)-KT-8110 with 5R,6S-configuration can be synthesized through oxidation. In addition a short route to (+)-bromoxone is described. Most steps include simple epoxide formation and cleavage reactions which all can be carried out in a high stereoselective manner. PMID- 10814006 TI - A computational study of nicotine conformations in the gas phase and in water. AB - The conformational preferences of nicotine in three protonation states and in the gas phase as well as aqueous solution are investigated using several computational procedures. Conformational aspects emphasized are N-methyl stereochemistry, relative rotation of the pyridine and pyrrolidine rings, and pyrrolidine ring conformation. All methods consistently predicted that the N methyl trans species are most stable for all protonation states in both gas phase and in water. However, the cis/trans energy gap is significantly reduced in water. Additionally, the two pyridine ring rotamers, which are energetically equivalent in the gas phase, experience different solvation energies in water. PMID- 10814009 TI - Efficient kinetic resolution in the asymmetric hydrosilylation of imines of 3 substituted indanones and 4-substituted tetralones. AB - Kinetic resolution of the N-methyl imines of 3-substituted indanones and 4 substituted tetralones could be accomplished by hydrosilylation with a chiral titanocene catalyst. N-Methyl imines of 4-substituted tetralones were resolved to yield, after hydrolysis of the unreacted starting materials, ketones with high ee's and the amine products with high diastereomeric and enantiomeric purity. The utility of this process was demonstrated in the synthesis of sertraline. PMID- 10814011 TI - Tamandarins A and B: new cytotoxic depsipeptides from a Brazilian ascidian of the family Didemnidae. AB - The structures of two new, naturally occurring cytotoxic depsipeptides, tamandarins A and B (1 and 2), are presented. The tamandarins were isolated from an unidentified Brazilian marine ascidian of the family Didemnidae. The structures of the new cytotoxins were assigned by interpretation of FABMS data and by extensive 2D NMR analyses. The absolute configurations of the tamandarins were assigned by acid and alkaline hydrolysis to yield their corresponding amino acids, which were then analyzed as their Marfey derivatives. The cytotoxicity of tamandarin A (1) was evaluated against various human cancer cell lines and shown to be slightly more potent than didemnin B. A qualitative discussion of the conformation of tamandarin A (1) in solution, obtained from NMR J-value data, variable temperature experiments, and NOESY/ROESY data, is included. PMID- 10814012 TI - Total syntheses of (+/-)-anchinopeptolide D and (+/-)-cycloanchinopeptolide D. AB - The first synthesis of (+/-)-anchinopeptolide D (4) has been accomplished in seven steps in 10% overall yield from octopamine hydrochloride (17), N (Boc)glycine (16), and 5-amino-2-hydroxypentanoic acid (22). The key step is the aldol dimerization and hemiaminal formation of alpha-keto amide 26, which gives primarily protected anchinopeptolide D 27 under kinetically controlled conditions. Cycloanchinopeptolide D (31) has been prepared by the unprecedented head-to-head photodimerization of the two hydroxystyrylamides of 4 using the hydrophobic effect in water to force the two side chains into close proximity so that [2 + 2] cycloaddition is faster than trans to cis double bond isomerization. Coupling of amine 21 with pyroglutamic acid affords the naturally occurring tripeptide 35, which had been assigned glutamic acid structure 34. PMID- 10814013 TI - Direct stereoselective synthesis of beta-thiomannosides. AB - A highly diastereoselective synthesis of beta-thiomannopyranosides is described in which S-phenyl 2,3-di-O-benzyl-4, 6-O-benzylidene-1-deoxy-1-thia-alpha-D mannopyranoside S-oxide is treated with triflic anhydride and 2, 6-di-tert-butyl 4-methylpyridine in CH(2)Cl(2) at -78 degrees C leading to the formation of an intermediate alpha-mannosyl triflate. Addition of primary, secondary, or tertiary thiols then leads to the beta-thiomannosides, by an S(N)2-like displacement, in good yield and with excellent stereoselectivity. Deprotection is achieved either by Birch reduction or by Zemplen deacetylation, of the acetyl protected aglycons, followed by hydrogenolysis over Pearlman's catalyst. The assignment of configuration of the beta-thiomannopyranosides is discussed in terms of the chemical shift of the mannose H5 resonance and the (1)J(CH) of the mannose anomeric carbon. PMID- 10814018 TI - First diastereoselective synthesis of (-)-methyl thyrsiflorin A, (-)-methyl thyrsiflorin B acetate, and (-)-thyrsiflorin C. AB - An efficient procedure for the synthesis of scopadulan diterpenes, using (+) podocarp-8(14)-en-13-one 13 as starting material, is reported. This procedure has been used for the diastereoselective synthesis of (-)-methyl thyrsiflorin A (8), (-)-methyl thyrsiflorin B acetate (9), and (-)-thyrsiflorin C (7). Key steps in our strategy are the intramolecular cyclopropanation of diazoketone 19 and the regioselective cleavage of the cyclopropane ring. PMID- 10814019 TI - Enzyme-assisted asymmetric total synthesis of (-)-podophyllotoxin and (-) picropodophyllin. AB - Described is the first catalytic, asymmetric synthesis of (-)-podophyllotoxin and its C(2)-epimer, (-)-picropodophyllin. Asymmetry is achieved via the enzymatic desymmetrization of advanced meso diacetate 20, through PPL-mediated ester hydrolysis. A second key feature of the synthesis is the strategically late introduction of the highly oxygenated natural ring E through an arylcopper species. The successful implementation of this approach augers well for the introduction of other functionalized rings E for future SAR work. The synthesis begins from piperonal, which is fashioned into isobenzofuran (IBF) precursor 14 in three steps (bromination, acetalization, and halogen-metal exchange/hydroxymethylation). Interestingly, treatment of 14 with HOAc in commerical dimethyl maleate (contains 5% dimethyl fumarate) leads to a nearly equimolar mixture of fumarate- (15) and maleate-IBF Diels-Alder adducts (16 and 17), indicating that IBF 11 reacts about 15 times faster with dimethyl fumarate than with dimethyl maleate. With scrupulously pure dimethyl maleate a 2.8:1 endo:exo mixture of maleate DA adducts is still obtained. On the other hand, the desired meso diester 16 is obtained pure and in nearly quantitative yield by employing neat dimethyl acetylene dicarboxylate as the dienophile, followed by catalytic hydrogenation. Reduction (LiAlH(4)) of 16 provides meso diol 19, which is then treated with Ac(2)O, BzCl, and PhCH(2)COCl to provide the corresponding meso diesters, 20-22. Screening of these meso benzoxabicyclo[2.2.1]heptyl substrate candidates across a battery of acyl transfer enzymes leads to an optimized match of diacetate 20 with PPL. Even on 10-20 g scales, asymmetry is efficiently introduced here, yielding the key chiral intermediate, monoacetate 25 (66% isolated yield, 83% corrected yield, 95% ee). Protecting group manipulation and oxidation (Swern) provide aldehyde 27b, which undergoes efficient retro Michael ring opening to produce dihydronaphthalene 30, in which the C(3) and C(4) stereocenters are properly set. Following several unsuccessful approaches to the intramolecular delivery of ring E (via Claisen rearrangement, Heck-type cyclization, or radical cyclization), a highly diastereoselective, intermolecular conjugate addition of the arylcopper reagent derived from (3,4,5 trimethoxy)phenylmagnesium bromide and CuCN to acyl oxazolidinone 50 was developed (85% yield, only the required alpha-stereochemistry at C(1) is observed). The conjugate addition product is converted to (-)-picropodophyllin in two steps (lactonization, SEM deprotection) or to (-)-podophyllotoxin, in three steps, through the introduction of a C(2)-epimerization step, under Kende conditions, prior to the final conjugate addition. PMID- 10814022 TI - Biocatalytic asymmetric hydroxylation of hydrocarbons with the topsoil microorganism Bacillus megaterium. AB - A Bacillus megaterium strain was isolated from topsoil by a selective screening procedure with allylbenzene as a xenobiotic substrate. This strain performed the hydroxylation chemoselectively (no arene oxidation and overoxidized products) and enantioselectively (up to 99% ee) in the benzylic and nonbenzylic positions of a variety of unfunctionalized arylalkanes. Salycilate and phenobarbital, which are potent inducers of cytochrome P-450 activity, changed the regioselectivity of the microbial CH insertion, without an effect on the enantioselectivity. The biotransformation conditions were optimized in regard to product yield and enantioselectivity by variation of the oxygen-gas supply and the time of the substrate addition. The different product distributions (alpha- versus beta hydroxylated product) that are obtained on induction of cytochrome P-450 enzyme activity demonstrate the involvement of two or more hydroxylating enzymes with distinct regioselectivities in this biotransformation. An oxygen-rebound mechanism is assumed for the cytochrome P-450-type monooxygenase activity, in which steric interactions between the substrate and the enzyme determine the preferred face of the hydroxy-group transfer to the radical intermediate. PMID- 10814025 TI - 13C and (2)H kinetic isotope effects and the mechanism of Lewis acid-catalyzed ene reactions of formaldehyde. AB - 13C and (2)H kinetic isotope effects were determined for the ene reaction of formaldehyde with 2-methyl-2-butene at natural abundance catalyzed by diethylaluminum chloride. The reactive methyl group exhibits a k(12)(C)/k(13)(C) of 1.006-1.009 and a k(H)/k(D) of approximately 1.22-1.23. The latter represents a combination of primary and secondary effects and is consistent with a significant primary deuterium isotope effect. A very close correspondence of the other isotope effects with the equilibrium isotope effects predicted for formation of a model intermediate cation is observed. An intermolecular deuterium isotope effect of 2.0-2.5 was observed under several reaction conditions in the Lewis acid-catalyzed reaction of formaldehyde with d(0)/d(12) tetramethylethylene. The results are interpreted as supporting the reversible formation of an essentially classical open cation followed by rate-limiting proton transfer. PMID- 10814029 TI - Synthesis of a chiral spiranic aminochroman derivative from L-proline. PMID- 10814034 TI - A new approach for the generation and reaction of organotin hydrides: the development of reactions catalytic in Tin PMID- 10814033 TI - 1,3-Calix PMID- 10814035 TI - A new thiatriazine isomer: synthesis, tautomerism, and molecular structure of 3,6 diphenyl-4H-1,2,4,5-thiatriazine as a precursor to the 1,2,4,5-thiatriazinyl radical AB - The diphenyl derivative of 4H-1,2,4,5-thiatriazine (5) was prepared by oxidative cyclization of 9. The molecular structure of 5, obtained by X-ray diffraction [orthorhombic, Pna2(1), a = 9.7746(13) A, b = 21.692(2) A, c = 5.6580(8) A], compares favorably with that predicted with ab initio calculations. The thiatriazine 5 was used as a precursor to the 3,6-diphenyl-1,2,4,5-thiatriazinyl radical (4) through either oxidation with PbO(2), or conversion to and reduction of sulfiminyl chloride 6 with Ph(3)Sb. The weak ESR quintet (a(N) = 1.03 mT, g = 2.0103) observed in the latter case correlates well with the molecular structure of 4, but the results of DFT calculations are ambiguous. Ab initio calculations show that 4H-1,2, 4,5-thiatriazine (I-4H) is the most stable tautomer and is the second most stable isomer among the six possible thiatriazines. All isomeric thiatriazinyl radicals exhibit similar spin distribution patterns. 1,2,4,5 Thiatriazinyl radical (I-R) is calculated to be 23. 1 kcal/mol less stable than the most stable 1,2,4,6 isomer II-R. PMID- 10814036 TI - Specific and chemoselective multi-alpha-arylation reaction of benzoylformic acid with or without decarbonylation in P(2)O(5)-MsOH and related acidic media AB - In P(2)O(5)-MsOH, or related acidic media, benzoylformic acid (1) undergoes three types of di- or mono-alpha-arylation reactions with or without decarbonylation ((1) decarbonylative alpha, alpha-diarylation, yielding triarylmethanols 6, (2) decarbonylative alpha-monoarylation, giving benzophenone derivatives 7, and (3) alpha,alpha-diarylation without decarbonylation, affording diarylated carboxylic acids 5) and one simple decarbonylation, without arylation, to form benzoic acid (8), instead of the conventional Friedel-Crafts acylation type reaction. The product ratios are governed by the capability of the acidic medium to form mixed anhydrides with carboxylic acids and the ability of the arenes to accept electrophiles. PMID- 10814037 TI - Reaction of zirconacycles with 3-iodopropenoates and 3-iodocycloenones in the presence of CuCl: A new pathway for the formation of cyclopentadienes and spirocyclic compounds AB - Formation of cyclic compounds from zirconacycles has been performed by a combination of Michael addition and coupling with an alkenyl iodide moiety in the presence of a stoichiometric amount of CuCl. The reaction of 3-iodopropenoates with various zirconacyclopentadienes in the presence of a stoichiometric amount of CuCl afforded penta- and hexasubstituted cyclopentadienes. The reaction of 3 iodocycloenones with zirconacyclopentadienes, zirconacyclopentenes, or zirconacyclopentanes gave spirocyclic compounds in good yields. PMID- 10814038 TI - Mechanism of phosphorus-carbon bond cleavage by lithium in tertiary phosphines. An optimized synthesis of 1,2-Bis(phenylphosphino)ethane AB - Conditions influencing the extent of P-C(aryl) vs P-C(alkyl) bond cleavage in the reaction of Ph(2)P(CH(2))(2)PPh(2) with lithium in THF have been investigated. The results complement and elucidate earlier work; they indicate that the mechanism of P-C bond cleavage in tertiary phosphines of this type involves a thermodynamic equilibrium between P-C(aryl) and P-C(alkyl) cleaved radicals and anions, followed by reaction and stabilization of these as lithium salts. The addition of water to the reaction mixture causes a reestablishment of the cleavage equilibrium prior to the formation of the secondary phosphines. A mechanism involving competitive release of leaving groups as the thermodynamically most stable anion or radical has been proposed. The preparation of (R, R)-(+/-)/(R, S)-PhP(H)(CH(2))(2)P(H)Ph by this route has been optimized. PMID- 10814039 TI - Use of n-pentenyl glycosides as precursors to various spacer functionalities. AB - Pent-4-enyl beta-D-glucopyranoside and its peracetylated and perbenzylated derivatives are shown to be excellent substrates for preparation of a wide variety of spacer functionalities. The spacer derivatives so obtained are promising substrates for preparing agents such as neo-glycoconjugates, micelles, and liquid crystalline phases, which are of interest for studying various biological and physiological phenomena of carbohydrates. PMID- 10814040 TI - C(5)H(7)O(2)(+) ions: the correlation between their thermochemistry in acidic solution and their chemistry in the gas phase AB - Each of a series of C(5)H(6)O(2) isomeric carboxylic acid and unsaturated lactones (1-7) was protonated in both concentrated sulfuric acid and trifluoromethanesulfonic acid. The thermally induced transformations of the protonated species were then studied over a temperature range of -30 to +160 degrees C. In the case of alpha,beta-unsaturated lactones, protonation took place on the carbonyl oxygen and gave the corresponding stable O-protonated species. Conversely, unconjugated lactones and acetylenic acid 7 were converted even at low temperature into the diprotonated ketoacid 8H(2)()o(+2)() by the acid catalyzed addition of water to the C-protonated precursor. Upon being heated at 160 degrees C, this acid gave protonated 1,3-cyclopentanedione. In the absence of water, decarbonylation followed by polymerization was observed in lactones 4 and 5. The CIMS spectra of compounds 1-7 were recorded using methane, ammonia, and moist air as reagent gases to determine the correlation between the fragmentation routes in the gas phase and the transformations observed in solution. Ammonia and moist air enabled us to determine the different proton affinities of these compounds. The data obtained in strong acids were used to assign reasonable structures to the gas-phase ions. PMID- 10814041 TI - Enthalpic and entropic contributions to the conformational free energies of methylthio, methylsulfinyl, methylsulfonyl, phenylthio, phenylsulfinyl, and phenylsulfonyl AB - A variable-temperature NMR study of (cis-4-methylcyclohexyl)methyl sulfide (1), sulfoxide (2), and sulfone (3), as well as (cis-4-methylcyclohexyl)phenyl sulfide (4), sulfoxide (5), and sulfone (6) allowed determination of the thermodynamic parameters, DeltaH degrees and DeltaS degrees, for the title groups. Reproduction of the experimental results with Allinger's MM3 program was successfully accomplished in the case of the sulfoxide and sulfide groups. Nevertheless, modification of the original force field torsional parameters was required in order to adequately reproduce the experimentally observed behavior of the sulfonyl derivatives. Rationalization of the enthalpic and entropic contributions to DeltaG degrees [S(O)(n)()R, n = 0, 1, 2; R = CH(3), Ph] is advanced in terms of the steric characteristics of these sulfur-containing groups and the resulting rotameric populations in the axial and equatorial monosubstituted cyclohexanes. PMID- 10814042 TI - Enhanced selectivities for the hydroxyl-directed methanolysis of esters using the 2-acyl-4-aminopyridine class of acyl transfer catalysts: ketones as binding sites. AB - In this paper we describe the preparation of a series of 2-acyl-4-aminopyridines, and their use as catalysts for the hydroxyl-directed methanolysis of alpha hydroxy esters in preference to alpha-methoxy esters. Hydroxyl-direction with these catalysts, which contain ketones at the 2-position of the pyridine, is achieved by reversible addition of the alcohol of the hydroxy ester to the ketone to provide the corresponding hemiketal. Their activity is compared to that of the previously described catalyst 2-formyl-4-pyrrolidinopyridine (FPP), which contains an aldehyde at the 2-position of the pyridine. The catalysts which contain ketones at the 2-position range in reactivity from 10 times slower to slightly faster than FPP, and certain of these are much more selective for the methanolysis of hydroxy esters than FPP. This increase in selectivity is ascribed to a decrease in the rate of the nondirected methanolysis reaction with the ketone-derived catalysts. The evidence suggests that the nondirected reaction does not proceed by an intermolecular general base mechanism, but rather via a nucleophilic catalysis mechanism in which the hydroxyl group of the hemiacetal formed upon addition of methanol to the aldehyde of FPP acts as the nucleophile. Since the hydroxyl group derived from a hemiketal is more hindered and less nucleophilic than that derived from a hemiacetal, the nondirected reaction is much slower for the catalysts containing ketones as binding sites. PMID- 10814043 TI - Synthesis of 2-(N-Acetylamino)-2-deoxy-C-glucopyranosyl nucleosides as potential inhibitors of chitin synthases. AB - The C-glucopyranosyl nucleosides (1-4) containing the N-acetyl glucosaminyl and uridine units have been synthesized as nonhydrolyzable substrate analogues of UDP GlcNAc aimed to inhibit the chitin synthases. The key intermediate, 4-(2'-(N acetylamino)-3', 4',6'-tri-O-benzyl-2'-deoxy-alpha-D-glucopyranosyl)but-2-enoic acid (5), was prepared from the perbenzylated (N-acetylamino)-alpha-C allylglucoside (7), by successive oxidative cleavage, Wittig olefination, and ester deprotection. The coupling of the acid 5 with the hydroxyl or amine function of the uridine derivatives (6a or 6b) afforded, respectively, the ester 12 and amide 14. The dihydroxylation of the conjugated double bond in ester 12 or amide 14 was better achieved with osmium tetraoxide/barium chlorate, leading to the expected diols 13 and 15 as a mixture of two diastereoisomers. The desired compounds 1-4 were obtained after catalytic hydrogenation of compounds 12-15. PMID- 10814044 TI - Novel acyl alpha-pyronoids, dictyopyrone A, B, and C, from Dictyostelium cellular slime molds. AB - For the elucidation of the diversity of secondary metabolites of Dictyostelium cellular slime molds, we investigate the constituent of three species of slime molds. From the methanol extract of their fruit bodies, we obtained three novel compounds, dictyopyrone A (1) and B (2) from D. discoideum and D. rhizoposium and dictyopyrone C (3) from D. longosporum. They possess a unique alpha-pyrone moiety with a side chain at the C-3 position. Their relative structures were elucidated by spectral means, and the absolute configuration was confirmed by asymmetric synthesis of 1. Since these compounds were obtained from different species of Dictyostelium slime molds, they may be a type of compound common to this genus. PMID- 10814045 TI - Structures of TMC-95A-D: novel proteasome inhibitors from Apiospora montagnei sacc. TC 1093. AB - Four novel proteasome inhibitors, TMC-95A-D (1-4) have been isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093, isolated from a soil sample. All of the molecular formulas of 1-4 were established as C(33)H(38)N(6)O(10) by high-resolution FAB-MS. Their planar structures were determined on the basis of extensive analyses of 1D and 2D NMR, and degradation studies. Compounds 1-4 have the same planar structures to each other, and are unique highly modified cyclic peptides containing L-tyrosine, L-aspargine, highly oxidized L-tryptophan, (Z)-1-propenylamine, and 3-methyl-2-oxopentanoic acid units. The absolute configuration at C-11 and C-36 of 1-4 was determined based on chiral TLC and HPLC analyses of their chemical degradation products. The ROESY analysis along with (1)H-(1)H coupling constants clarified the absolute stereochemistry at C-6, -7, -8, and -14 of the cyclic moieties. These studies revealed the relationships of 1-4 to be diastereomers at C-7 and C-36. PMID- 10814046 TI - Chemo- and stereoselective monobenzoylation of 1,2-diols catalyzed by organotin compounds AB - A new facile method for monoacylation of diols has been developed. A variety of cyclic and acyclic diols, in particular 1,2-diols, were selectively monobenzoylated in good yields by the reaction with benzoyl chloride in the presence of a catalytic amount of dimethyltin dichloride and inorganic bases such as potassium carbonate. Furthermore, the method was successfully applied to a kinetic resolution of racemic 1-phenyl-1,2-ethanediol using a chiral organotin catalyst. The ee was dependent on the kind of base, water as an additive, and the reaction temperature. PMID- 10814047 TI - A novel route to 5-substituted 3-isoxazolols. Cyclization of N, O-DiBoc beta-keto hydroxamic acids synthesized via acyl Meldrum's acids. AB - 3-Isoxazolols are most often synthesized from a beta-keto ester and hydroxylamine. This cyclization typically gives rise to a major byproduct, the corresponding 5-isoxazolone. We have found that N, O-diBoc-protected beta-keto hydroxamic acids can be synthesized and cyclized to 5-substituted 3-isoxazolols without formation of any byproduct. We present a novel and versatile three-step procedure in which carboxylic acid derivatives are converted into acyl Meldrum's acids which, upon aminolysis with N, O-bis(tert-butoxycarbonyl)hydroxylamine, lead to the N, O-diBoc-protected beta-keto hydroxamic acids. These hydroxamic acid analogues were then, upon treatment with hydrochloric acid, cyclized to the corresponding 5-substituted 3-isoxazolols. PMID- 10814048 TI - Reduction of permanganate by thioanisole: lewis acid catalysis AB - The oxidation of sulfides and sulfoxides by permanganate in anhydrous acetone solutions is catalyzed by Lewis acids such as iron(III) chloride, zinc chloride, and mercury(II) chloride. The reaction kinetics unequivocally confirm that the function of these catalysts is to activate the oxidant by forming permanganate/Lewis acid complexes analogous to the protonation of Mn by Bronsted acids. A Hammett analysis of the rate constants for the oxidation of a series of substituted thioanisoles gives a negative rho value (-1. 11) indicative of an electron deficient transition state. No secondary kinetic isotope effect is observed when the hydrogens of the methyl group are replaced by deuterium. Despite previous observations that sulfoxides are preferentially oxidized in competitive experiments, sulfides are oxidized more rapidly when individual rates are measured. All of these observations are most consistent with a mechanism in which the reductant reacts with the oxidant via initial ligation. PMID- 10814049 TI - Mechanistic studies of a linear trisazoalkane, a new azimine, and a bicyclic triaziridine. Azoalkane homolysis into seven fragments AB - An aliphatic azo compound containing three azo groups (1) has been prepared by IF(5) oxidation of beta-azoamine 3. The thermolysis kinetics of this vicinal trisazoalkane were investigated above 155 degrees C, leading to a rate constant only 5.5 times faster than that of the simple model, azo-tert-butane. Because thermolysis to form seven stable products proceeds stepwise, the rate is hardly affected by the high exothermicity of the overall reaction (-93.4 kcal/mol). Oxidation of amine 3 also afforded a cyclic azimine 5 that underwent photolysis to yield a highly strained triaziridine 9 plus an unusual triazane 10, whose structures were elucidated by detailed NMR studies. On standing at ambient temperature, 9 reverted to 5 with a half-life of about an hour. PMID- 10814050 TI - Total synthesis of the fumiquinazoline alkaloids: solution-phase studies. AB - Biomimetic total syntheses of glyantrypine, fumiquinazoline F, fumiquinazoline G, and fiscalin B were achieved in four steps from tryptophan methyl ester. In the key step, the anthranilamide residue in a linear tripeptide is dehydrated to a benzoxazine by reaction with triphenylphosphine, iodine, and a tertiary amine. The benzoxazines subsequently undergo rearrangement to the natural products via an amidine intermediate. This dehydrative oxazine to quinazoline route is applicable to a broad range of N-acylanthranilamides, including sterically hindered cases. PMID- 10814051 TI - Nucleophilic additions of arylzinc compounds to aldehydes mediated by CrCl(3): efficient and facile synthesis of functionalized benzhydrols, 1(3H) isobenzofuranones, benzyl alcohols, or diaryl ketones AB - In the presence of a stoichiometric amount of CrCl(3) and trimethylchlorosilane (TMSCl), nucleophilic addition of arylzinc compounds 1c-h to arylaldehydes 2a,b,g smoothly proceeded at room temperature to yield corresponding benzhydrols 4a-f in good yields. From arylzinc compounds 1a,b, 3-aryl-1(3H)-isobenzofuranones 3a-f were given by the CrCl(3)-mediated reaction with arylaldehydes 2a-f. Diaryl ketones 5a-e were obtained in good yields by the addition of excess amount of benzaldehyde as an oxidant to the resulting solution after the CrCl(3)-mediated reaction between arylzinc compounds 1c-g and arylaldehydes 2b,g was completed. In the nucleophilic additions of arylzinc compounds 1a,d,f to alkyladehydes 6b-f, the treatment of arylzinc compounds with CrCl(3) was required prior to the addition of the aldehydes in order to prevent the fast protodezincation of arylzinc compounds by the enolizable aldehydes. In these CrCl(3)-mediated nucleophilic additions of arylzinc compounds to aldehydes, arylchromium(III) species are probably reactive intermediates. PMID- 10814052 TI - Total synthesis and revision of stereochemistry of the marine metabolite trunkamide A. AB - The isolation of the cytotoxic Lissoclinum sp. metabolite trunkamide A was reported in 1996. After completion of a total synthesis in 1999, it became clear that the structure of this marine natural product had to be revised. We now report the first preparation of actual trunkamide A in a total synthesis that serves as an unambiguous structural and stereochemical proof. Highlights of our synthetic strategy are a Lewis acid assisted aziridine opening that was used for the preparation of the novel reverse-prenylated serine and threonine side chains as well as an efficient oxazoline-thiazoline interconversion on the macrocyclic skeleton. In addition, several stereoisomers prepared by complementary synthetic protocols serve to illustrate the general scope of our methodology and confirm the configurational assignment. PMID- 10814053 TI - From homoleptic to heteroleptic double stranded Copper(I) helicates: the role of self-recognition in self-assembly processes AB - The ligands 2,9-bis[(6-methyl-2, 2'-bipyridin-6'-yl)methyleneoxymethylenyl]-1,10 phenanthroline (6), 6' ',6' "-bis[(6-methyl-2, 2'-bipyridin-6' yl)methyleneoxymethylenyl)]-2' ',2' "-bipyridine (2), 5,5'-bis[(6-methyl-2,2' bipyridin-6'yl)methyleneoxymethylenyl]-2, 2'-bithiophene (7), and 6,6'-bis[(6 methyl-2, 2'-bipyridin-6'-yl)methyleneoxymethylenyl]-2,2'-biphenyl (8) and their respective homo- and heteroleptic double-stranded copper(I) complexes were prepared and characterized in order to estimate the importance of self recognition in the self-assembly processes of double-stranded copper complexes. The homoleptic double-stranded copper complexes of 2, 6, 7, and 8 were characterized by NMR, FAB-MS, and electrochemistry. It was found that 6 and 2 each form a single double-stranded helicate having the structure of [(L)(2)Cu(3)](3+) (L = 2 or 6), 7 forms two double-stranded [(7)(2)Cu(3)](3+) complexes, and 8 results in a mixture of at least two [(8)(2)Cu(2)](2+) complexes. The potential shift, DeltaE degrees (,) of the Cu(+)/Cu(2+) redox process of these complexes reflects the binding affinity of the different binding sites to the copper cation. The electrochemical data show that the central units have a higher affinity to Cu(+) as compared to the off-center binding sites. NMR was used to determine the actual complex composition obtained from different mixtures of 2, 6, or 7 with Cu(+). Interestingly, we have found that, although 6, 2, and 7 each form homoleptic double-stranded complexes, no heteroleptic double stranded copper complexes were formed from the mixtures of 7 with either 6 or 2. However, when mixtures of 6 and 2 are used, helicate distributions seem to follow simple statistics. These results are discussed in terms of the relative importance of self-recognition in the self-assembly of double-stranded helicates. PMID- 10814054 TI - The oxetane ring in taxol. AB - Numerous structure-activity studies combining synthesis and bioassay have been performed for the anti-cancer drug Taxol. The four-membered D-ring, an oxetane, is one of four structural features regarded to be essential for biological activity. This proposition is examined by application of a Taxol-epothilone minireceptor, K(i) estimation for microtubule binding and docking of Taxol analogues into a model of the Taxol-tubulin complex. In this way, we evaluate the two characteristics considered responsible for oxetane function: (1) rigidification of the tetracyclic Taxol core to provide an appropriate framework for presenting the C-2, C-4, C-13 side chains to the microtubule protein and (2) service as a hydrogen-bond acceptor. An energy decomposition analysis for a series of Taxol analogues demonstrates that the oxetane ring clearly operates by both mechanisms. However, a broader analysis of four-membered ring containing compounds, C- and D-seco derivatives, and structures with no oxetane equivalent underscores that the four-membered ring is not necessary for Taxol analogue bioactivity. Other functional groups and ligand-protein binding characteristics are fully capable of delivering Taxol biobehavior as effectively as the oxetane D ring. This insight may contribute to the design and development of novel anticancer drugs. PMID- 10814055 TI - Halonium ion-mediated reaction of unsaturated hydroperoxy acetals. Competition between the formation of cyclic peroxides and the migration of the methoxy (or hydroxy) group. AB - Monoozonolyses of dienes 2 in methanol gave in each case the corresponding unsaturated alpha-methoxy hydroperoxides 3. Capture of 2-alkyl-substituted cyclohexanone oxides by methanol was highly diastereoselective, thereby providing exclusively the hydroperoxides derived from attack by methanol from the less hindered face of the carbonyl oxide intermediates. Halonium ion-mediated reactions of the hydroperoxides 3 gave the novel methoxy- or hydroxy-migrated products, together with the expected halogen-substituted 1, 2-dioxanes and/or 1,2 dioxepanes, the composition of the product mixture being a function of the halogenating agent utilized and the structure of 3. PMID- 10814056 TI - A DFT study of the domino inter AB - The molecular mechanism of the domino inter [4 + 2]/intra [3 + 2] cycloaddition reactions of nitroalkenes with enol ethers to give nitroso acetal adducts has been characterized using density functional theory methods with the B3LYP functional and the 6-31G basis set. The presence of Lewis acid catalyst and solvent effects has been taken into account to model the experimental environment. These domino processes comprise two consecutive cycloaddition reactions: the first one is an intermolecular [4 + 2] cycloaddition of the enol ether to the nitroalkene to give a nitronate intermediate, which then affords the final nitroso acetal adduct through an intramolecular [3 + 2] cycloaddition reaction. The intermolecular [4 + 2] cycloaddition can be considered as a nucleophilic attack of the enol ether to the conjugated position of the nitroalkene, with concomitant ring closure and without intervention of an intermediate. For this cycloaddition process, the presence of the Lewis acid favors the delocalization of the negative charge that is being transferred from the enol ether to the nitroalkene and decreases the activation energy of the first cycloaddition. The [4 + 2] cycloaddition presents a total regioselectivity, while the endo/exo stereoselectivity depends on the bulk of the Lewis acid used as catalyst. Thus, for small Lewis acid catalyst, modeled by BH(3), the addition presents an endo selectivity. The [3 + 2] cycloaddition reactions present an total exo selectivity, due to the constraints imposed by the tether. Inclusion of Lewis acid catalyst and solvent effects decrease clearly the barrier for the first [4 + 2] cycloaddition relative to the second [3 + 2] one. Calculations for the activation parameters along this domino reaction allow to validate the results obtained using the potential energy barriers. PMID- 10814057 TI - Efficient synthesis of monoacyl dipyrromethanes and their use in the preparation of sterically unhindered trans-porphyrins. AB - The condensation of an aldehyde with a dipyrromethane bearing a sterically unhindered aryl substituent at the 5-position typically results in low yield and a mixture of porphyrin products derived from acidolytic scrambling. We have developed a concise nonscrambling synthesis of such trans-porphyrins that takes advantage of the availability of multigram quantities of dipyrromethanes. This route involves the selective monoacylation of the dipyrromethanes with a pyridyl thioester, reduction of the monoacyl dipyrromethane to the corresponding carbinol, and self-condensation of the carbinol to form the porphyrin. The monoacylation procedure has wide scope as demonstrated by the preparation of a set of 15 diverse monoacyl dipyrromethanes in good yield at the multigram scale. The dipyrromethanecarbinol self-condensation reaction is extremely rapid (<3 min) under mild room-temperature conditions and affords the trans-porphyrin in 16-28% yield. Analysis by laser-desorption mass spectrometry (LD-MS) of samples from the crude reaction mixture revealed no scrambling within the limit of detection (1 part in 100). The self-condensation is compatible with a range of electron withdrawing or -releasing substituents as well as substituents for building block applications (TMS-ethyne, ethyne, iodo, ester). The absence of any detectable scrambling in the self-condensation enables a simple purification. The synthesis readily affords gram quantities of pure, sterically unhindered trans-porphyrins in a process involving minimal chromatography. PMID- 10814058 TI - Predicting efficient C(60) epoxidation and viable multiple oxide formation by theoretical study AB - The epoxidation of C(60) by various oxidizing agents such as dimethyldioxirane (DMD), methyl(trifluoromethyl)dioxirane (MTMD), and bis(trifluoromethyl)dioxirane (BTMD) has been probed computationally by the AM1 method. The computations have revealed that for the reaction forming C(60)O through a concerted "spiro" transition state, the currently used DMD involves its HOMO lone-pair and the LUMO (pi) of fullerene in an inverse electron demand fashion. This is distinct from the DMD reaction with ethylene. On the other hand, the addition of CF(3) groups lowers the LUMO (peroxide sigma) of MTMD and BTMD by virtue of negative hyperconjugation; the oxidants can then attack the fullerene nucleophilically at an increased rate to the maximum extent. These estimations have thus established that the strong electrophilic oxidizing agents remarkably enhance the fullerene epoxidation. DMD further produces C(60)O(2) and C(60)O(3) via multiple epoxidations, as C(60)O might best be produced quantitatively by MTMD and BTMD. The regiochemistry of the multiple oxidation products in which the subsequent oxidations take place at the adjacent sites is consistent with the increased nucleophilicity of the nearest double bonds attached to the prevailing epoxide function. PMID- 10814059 TI - On the mechanism of the dimethyldioxirane oxidation of sigma(H) adducts (Meisenheimer complexes) generated from nitroarenes and carbanions AB - The mechanism of the novel dimethyldioxirane (DMD) oxidation of sigma(H) adducts (Meisenheimer complexes) generated from nitroarenes and carbanions was elucidated. The proposed mechanism, which is akin to that of the oxidative Nef reaction, is supported by the isolation of the cyclohexadienone intermediate and the lack of a primary kinetic isotopic effect. Protic solvents (H(2)O, MeOH) enhance the reactivity of DMD through intermolecular hydrogen bonding. PMID- 10814060 TI - Design and synthesis of novel thiazole-containing cross-linked polyamides related to the antiviral antibiotic distamycin. AB - A family of naturally occurring oligopeptides includes netropsin, distamycin, anthelvencin, kikumycin B, amidinomycin, and norformycin. Netropsin (I) and distamycin (II) express their biological activities by targeting specific sequences of chemical functionalities in the minor groove of DNA. Both netropsin and distamycin can be regarded as polyamide chains in which each alpha-carbon has been replaced by a five-membered pyrrole ring. The repeat distance in such an augmented polyamide chain is almost the same as the distance from one base pair to the next along the floor of a minor groove within beta-DNA. In this paper we report the synthesis of 16-21 cross-linked polyamides containing a thiazole heterocyclic ring bearing the active functionalites NH(2), NHCHO, or H. 16 and 17 were synthesized by DCC and HOBt catalyzed reaction of 5 with 14 and 15, while the formylation products 18 and 19 were obtained by coupling the formylated 4 methyl-thiazolated acid 6 with 14 and 15. The deaminated compounds 20 and 21 were obtained by the coupling of 5-trichloroacetyl-4-methylthiazole 7 synthesized from 4-methylthiazole. All the six cross-linked polyamides 16-21 were tested for their DNA gyrase inhibition. The studies have shown these polyamides have better sequence recognition and a greater percentage of inhibition than the corresponding monomers. The compound 17 shows complete inhibition of gyrase at 0.5 microM concentration as compared to the naturally occurring distamycin at 1.0 microM. PMID- 10814061 TI - Synthesis of optically active 5-substituted-2-pyrrolidinone derivatives having atropisomeric structure and 3,5-cis-selective reaction of their enolates with electrophiles AB - Optically pure forms (>/=98% ee) of N-(o-tert-butylphenyl)-5-(methoxymethyl)-2 pyrrolidinone having atropisomerism and N-(o-tert-butyldiphenylsiloxyphenyl)-5 (methoxymethyl)-2-pyrrolidinon e having an atropisomerism-like structure were prepared from ortho-substituted aniline derivatives and (S)-5 (methoxymethyl)butyrolactone in a stereoselective manner. The reactions of Li enolates from these lactams with various electrophiles and subsequent dearylation of the products gave 3, 5-cis-disubstituted-2-pyrrolidinone derivatives. PMID- 10814062 TI - Reaction of essentially free benzyl cations with acetonitrile; synthesis of ethanimidic carboxylic anhydrides and unsymmetrical diacylamines AB - Benzyl cations were generated via the thermal decomposition of N-benzyl-N nitrosopivalamide in acetonitrile and acetonitrile-water mixtures at 25 degrees C. The first-formed (primary) benzylating agent, the benzyl cation, was scavenged competitively by pivalate (trimethyl acetate) ion to yield benzyl pivalate, by acetonitrile to yield the corresponding N-benzylnitrilium ion, and by water (when present) to yield benzyl alcohol. The nitrilium ion underwent a cascade of reactions to yield an array of products whose identities and relative yields as a function of time were used to elucidate the mechanistic paths involved. Thus, the N-benzylnitrilium ion reacted with pivalate ion to yield the Z-isomer of N benzylethanimidic pivalic anhydride, followed by its conversion into the E isomer. This article appears to be the first to document compounds of this type. The E-isomer is labile under the reaction conditions, rearranging into N-acetyl-N pivalylbenzylamine. In the presence of water as a diluent, a significant fraction of the nitrosoamide was hydrolyzed to benzyl alcohol; hydrolysis of the nitrilium ion yielded N-benzyl acetamide. The yield of hydrosylates was directly proportional to the mole fraction of water in the medium. PMID- 10814063 TI - A study of N-nitrosoamide-mediated friedel-crafts type benzylation of benzene toluene and benzene-Anisole(1)(a) AB - Nitrogen-separated carbocation-carboxylate ion pairs were employed as sources of carbocations in the alkylation of aromatic compounds. The N-nitrosoamide approach to these nitrogenous-molecule-separated ion pairs is an excellent alternative to the standard acid-catalyzed Friedel-Crafts approach for studies of the alkylation because of the following variables: high reactivity of the electrophile, stability of the products, strict kinetic control, homogeneity, lack of overalkylation, straightforward chemistry, and good product balance. In deaminative benzylations of benzene-toluene and anisole-benzene mixtures, the values of k(X)/k(B) and % meta isomer are significantly different from those observed in the standard benzylations in a manner that indicates the deaminative electrophiles are more reactive than those generated by the standard Friedel Crafts approach. The reactions show a direct proportionality between inter- and intramolecular selectivities and thus follow the Brown selectivity relationship (BSR). The benzylation of 2,4, 6-trideuteriotoluene provided no evidence for deuterium rearrangements (or coupled benzyl rearrangements) in the arenium ion intermediate. Kinetic isotope effects were not detected. The methyl substituent on toluene appears to affect intermolecular selectivity (k(T)/k(B)) and intramolecular selectivity (o, m, p distribution) to similar degrees. A mechanism is proposed in which both selectivities are determined by activation energy differences in the transition states leading to the sigma-complex intermediates. The observation that most standard Friedel-Crafts benzylations do not follow the BSR is discussed in terms of possible systematic errors in those cases. Silver ion-assisted Friedel-Crafts benzylations were performed under dry, basic conditions to investigate whether the standard approach could lead to data that obey the BSR. PMID- 10814064 TI - Functional-group-directed diastereoselective hydrogenation of aromatic compounds. 2(1) AB - Diastereoselective liquid-phase hydrogenation of a series of monosubstituted indan substrates was studied on supported rhodium catalysts. Predominantly the cis-cis diastereomer, obtained by hydrogenation from the diastereoface opposite the substituent at the stereogenic center, and the cis-trans diastereomer, obtained by hydrogenation from the diastereoface on the same side as the substituent, were formed. The diastereoselectivity depends on the balance between steric repulsion and electronic attraction of the substituent with the surface of the catalyst. For alkoxy and carboxyl groups (acid, methyl ester, and amide), the steric repulsion dominated and the cis-cis diastereomer was obtained with moderately high selectivity. The diastereoselectivity obtained in the hydrogenation was influenced by the addition of bases to the reaction mixture. Addition of triethylamine caused a small increase in the selectivity to the cis cis diastereomer in some substrates, whereas the addition of NaOH significantly increased the selectivity toward the cis-trans isomer in all substrates. PMID- 10814065 TI - Nucleophilic addition of hydroxylamine, methoxylamine, and hydrazine to malononitrileoxime AB - The chemistry of malononitrileoxime, HONC(CN)(2), with respect to nucleophilic addition to ammonia, methylamine, hydroxylamine, methoxylamine, and hydrazine is reported. Whereas the poorly nucleophilic ammonia and methylamine do not react, hydroxylamine, methoxylamine, and hydrazine add to the nitrile groups of the oxime, yielding the corresponding amidoximes and amidrazones. Depending on the stoichiometry of the reactions, hydroxylamine and hydrazine add to one or both of the nitrile groups; methoxylamine adds to only one of the nitrile groups. Three of the products, namely, cyanoacetamidoxime (1), 3-amino-2,3 hydroxyiminopropionitrile monohydrate (2.H(2)O), and 3,5-diaminopyrazolone-4 oxime monohydrochloride monohydrate (6.HCl.H(2)O), are characterized by single crystal X-ray diffraction data. All of the products exhibit exothermic decomposition properties with heats of decomposition in the range of 500-1500 kJ mol(-)(1). PMID- 10814066 TI - Scope and limitations of the Pd/BINAP-catalyzed amination of aryl bromides. AB - Mixtures of Pd(2)(dba)(3) or Pd(OAc)(2) and BINAP catalyze the cross-coupling of amines with a variety of aryl bromides. Primary amines are arylated in high yield, and certain classes of secondary amines are also effectively transformed. The process tolerates the presence of several functional groups including methyl and ethyl esters, enolizable ketones, and nitro groups provided that cesium carbonate is employed as the base. Most reactions proceed to completion with 0.5 1.0 mol % of the palladium catalyst; in some cases, catalyst levels as low as 0.05 mol % Pd may be employed. Reactions are considerably faster if Pd(OAc)(2) is employed as the precatalyst, and the order in which reagents are added to the reaction has a substantial effect on reaction rate. It is likely that the catalytic process proceeds via bis(phosphine)palladium complexes as intermediates. These complexes are less prone to undergo undesirable side reactions which lead to diminished yields or catalyst deactivation than complexes of the corresponding monodentate triarylphosphines. PMID- 10814067 TI - Simple, efficient catalyst system for the palladium-catalyzed amination of aryl chlorides, bromides, and triflates. AB - Palladium complexes supported by (o-biphenyl)P(t-Bu)(2) (3) or (o-biphenyl)PCy(2) (4) are efficient catalysts for the catalytic amination of a wide variety of aryl halides and triflates. Use of ligand 3 allows for the room-temperature catalytic amination of many aryl chloride, bromide, and triflate substrates, while ligand 4 is effective for the amination of functionalized substrates or reactions of acyclic secondary amines. The catalysts perform well for a large number of different substrate combinations at 80-110 degrees C, including chloropyridines and functionalized aryl halides and triflates using 0.5-1.0 mol % Pd; some reactions proceed efficiently at low catalyst levels (0.05 mol % Pd). These ligands are effective for almost all substrate combinations that have been previously reported with various other ligands, and they represent the most generally effective catalyst system reported to date. Ligands 3 and 4 are air stable, crystalline solids that are commercially available. Their effectiveness is believed to be due to a combination of steric and electronic properties that promote oxidative addition, Pd-N bond formation, and reductive elimination. PMID- 10814068 TI - Generation of the enol of methyl mandelate by flash photolysis of methyl phenyldiazoacetate in aqueous solution and study of rates of ketonization of this enol in that medium. AB - Flash photolysis of methyl phenyldiazoacetate in aqueous solution produced phenylcarbomethoxycarbene, whose hydration generated a short-lived transient species that was identified as the enol isomer of methyl mandelate. This assignment is supported by the shape of the rate profile for decay of the enol transient, through ketonization to its carbonyl isomer, as well as by solvent isotope effects and the form of acid-base catalysis of the ketonization reaction. Comparison of the present results with previously published information on the enol of mandelic acid shows some interesting and readily understandable similarities and differences. PMID- 10814069 TI - Conformational studies in the cyclohexane series. 2. Phenylcyclohexane and 1 methyl-1-phenylcyclohexane AB - The structures and relative energies of the conformers of phenylcyclohexane, and 1-methyl-1-phenylcyclohexane have been calculated at theoretical levels including HF/6-31G, B3LYP/6-311G, MP2/6-311G, MP2/6-311(2df,p), QCISD/6-311G, and QCISD/6 311G(2df,p). The latter gives conformational enthalpy (DeltaH degrees ), entropy (DeltaS degrees ), and free energy (DeltaG degrees ) values for phenylcyclohexane that are in excellent agreement with the experimental data. The calculations for 1-methyl-1-phenylcyclohexane find a free energy difference of 1.0 kcal/mol at 100 degrees C, favoring the conformation having an axial phenyl group, that is in only modest agreement with the experimental value of 0.32 +/- 0.04 kcal/mol. The origin of the phenyl rotational profiles for the conformers of phenylcyclohexane and 1-methyl-1-phenylcyclohexane is discussed. PMID- 10814070 TI - Use of competition kinetics with fast reactions of grignard reagents AB - Competition kinetics are useful for estimation of the reactivities of Grignard reagents if the reaction rates do not differ widely and if exact rates are not needed. If the rate of mixing is slower than the rate of reaction, the ratios between the rates of fast and slow reagents are found to be too small. This is concluded from experiments in which results obtained by competition kinetics are compared with results obtained directly by flow stream procedures. A clearer picture of the reactivity ratios is obtained when the highly reactive reagent is highly diluted with its competitor. A fast reagent may account for almost all the product even when present as only 1 part in 100 parts of the competing agent. In this way allylmagnesium bromide is estimated to react with acetone, benzophenone, benzaldehyde, and diethylacetaldehyde ca. 1.5 x 10(5) times faster than does butylmagnesium bromide. The rates found for the four substrates do not differ significantly, and it seems possible that there is a ceiling over the rate of reaction of this reagent, for example, caused by diffusion control. This may explain that competition kinetics using allylmagnesium bromide have failed to show kinetic isotope effects or effects of polar substituents with isotopically or otherwise substituted benzophenones. A recently reported alpha-deuterium secondary kinetic isotope effect for the reaction of benzaldehyde with allylmagnesium bromide was observed at -78 degrees C, but was absent at room temperature. It is suggested that the reaction of benzophenone and benzaldehyde with allylmagnesium bromide has a radical-concerted mechanism since no radical type products are produced and since no color from an intermediate ketyl is observed even at -78 degrees C. PMID- 10814071 TI - Fixation of heterocumulenes. 1. A theoretical study on the irreversible reaction of CO(2) with a 2-lithio-1,3-dithiane AB - The mechanism of electrophilic addition of CO(2) to 2-lithio-2-phenyl-1, 3 dithiane-tetrahydrofuran-tetramethylethylendiamine, a Corey-Seebach "umpolungs" reagent for nucleophilic acylation, was investigated at the B3YLP/6-311+G//HF/6 31+G level of theory with specific solvation effects being included in the study. The overall reaction is exothermic by 18.8 kcal/mol and proceeds via precomplexation of the CO(2) (necessary for activation). The reaction barrier is calculated to be 11.9 kcal/mol (relative to the precomplex 2a) and represents a lower boundary for the activation energy. The reaction barrier is shown to originate from specific solvent effects. A detailed survey of the electronic effects governing the reaction is given. The ab initio results were then compared with semiempirical PM3 calculations, which were extended to include the heterocumulenes COS and CS(2). As expected, the reduced electrophilicity of these molecules leads to a higher activation barrier by retention of the same reaction mechanism. PMID- 10814072 TI - Synthesis and characterization of 5-substituted 1,3-diazacyclohexane derivatives AB - Three synthetic routes to 5-substituted 1,3-diazacyclohexane derivatives 1 are reported. The first method involves treatment of 1, 3-diaminopropan-2-ol 2 with paraformaldehyde to yield 5-hydroxy-1, 3-diazacyclohexane 3. A second method is based on the condensation of 2-bromo-2-nitro-1,3-propanediol with tert-butylamine and formaldehyde to yield 1,3-di-tert-butyl-5-bromo-5-nitro-1, 3-diazacyclohexane 22. The third method relies on the cycloalkylation of methylenebisacetamide with 3-chloro-2-chloromethyl-2-propene to provide 5-exomethylene-1, 3-diacetyl-1,3 diazacyclohexane 28. Functional group manipulations of 3, 22, and 28 provide a number of novel 1,3-diazacyclohexanes functionalized at the 5-position. PMID- 10814073 TI - Convenient acylative dealkylation of tertiary amines PMID- 10814074 TI - Microwave-induced esterification using heterogeneous acid catalyst in a low dielectric constant medium PMID- 10814075 TI - Hetero diels-alder reactions catalyzed by the metallocenium complex [Cp(2)Ce][BPh(4)]. PMID- 10814076 TI - Design of novel derivatives of phosphonoformate (Foscarnet) as prodrugs and antiviral agents. PMID- 10814077 TI - Asymmetric synthesis of a C-3 substituted pipecolic acid. PMID- 10814078 TI - Synthesis and reactivity of pyridylpyridone derivatives. PMID- 10814080 TI - Anodic oxidation of N-protected 4-methoxy anilines: improved synthesis of quinone imine acetals PMID- 10814079 TI - Sequential acyl halide-aldehyde cyclocondensation and enzymatic resolution as a route to enantiomerically enriched beta-lactones. PMID- 10814081 TI - Nozaki-hiyama reactions on halogenated allylchromium reagents: A new entry for the preparation of quaternary halogenated carbons PMID- 10814082 TI - Elimination reactions of aryl phenylacetates promoted by R(2)NH/R(2)NH(2)(+) in 70 mol MeCN(aq). Effect Of the beta-phenyl group on the ketene-forming transition state PMID- 10814083 TI - (+)-(18-Crown-6)-2,3,11,12-tetracarboxylic acid and its Ytterbium(III) complex as chiral NMR discriminating agents AB - The compound (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (I) and its ytterbium(III) complex are evaluated as chiral NMR discriminating agents. The crown ether is a useful chiral discriminating agent for protonated amino acid esters, amines, and amino alcohols. The crown can also be used with neutral primary amines since amines are protonated through a neutralization reaction with a carboxylic acid moiety of the crown. Enantiodiscrimination with the crown is observed in methanol and acetonitrile. Addition of ytterbium(III) nitrate to crown-substrate mixtures causes upfield shifts in the NMR spectrum of the substrate and often enhances the enantiomeric discrimination. Evidence indicates that the ytterbium(III) bonds to the carboxylic acid moieties of the crown, but enhancements in enantiomeric discrimination result from either the different association constants of the enantiomers with the crown or diastereomeric nature of the resulting crown-substrate complexes. The ytterbium complex with the crown is suitable for use in methanol but precipitates in acetonitrile. PMID- 10814084 TI - Dearomatization of naphthalene: novel stereoselective cyclization reactions promoted by Osmium(II) AB - A series of Michael acceptors has been combined with the Os(II) eta(2) naphthalene complex (1) to form stable 1H-naphthalenium species. Under acidic conditions, these complexes undergo ring closure at C2 to form the phenanthrenone core. In contrast, the corresponding 1-methylnaphthalene complex (15) upon addition of MVK at C8 undergoes ring closure at C5 to form a bridged tricyclic complex (18). Michael addition of MVK to the naphthalene complex (1) followed by deprotonation, an inter-ring linkage isomerization, and ring closure forms a 9 methylphenalene complex (21). In all cases, the organic cyclization products may be decomplexed by heating with silver triflate and isolated in moderate yield. PMID- 10814085 TI - A synthesis of conjugatively bridged bis- and tris-5-(2, 2'-Bipyridines): multitopic metal ion-binding modules for supramolecular nanoengineering AB - An efficient preparation of linear and curved bis- and branched tris-5-(2,2' bipyridines) of nanoscopic dimensions possessing rigid conjugated bridges is presented. The synthesis, which avoids the need of protection/deprotection methodology, utilizes central bridge precursors which are outwardly di- and trifunctionalized with a 5-(2-chloropyridine) synthon via a chemoselective palladium-catalyzed Sonogashira or Negishi cross-coupling protocol to yield the bridged linear (5a-c, 5f,g) and curved (6, 7) bis- and branched (8) tris-5-(2 chloropyridines). Under more forcing conditions, the ethyne-bridged 5-(2 chloropyridines) undergo the Stille cross-coupling reacton with 2 trimethylstannylpyridines to afford the conjugatively bridged linear (1a,b, 1g-j) and curved (2a, b, 3a,b) bis- and branched, (4a,b) tris-5-(2,2'-bipyridines) in good overall yields. The phenyl- and biphenyl-bridged linear bis-5-(2, 2' bipyridines) (1c-f) were best prepared from the bis-5-(2-bromopyridines) (5d,e) to ensure completion of the Stille cross-coupling reactions. The Stille cross couplings showed a marked substituent effect in which the terminally phenylated bis- and tris-5-(2,2'-bipyridines) were formed in higher yields than the methyl substituted analogues with the same bridge. The advantages of the methodology lie in its synthetic convenience and adaptibility for creating multitopic metal ion binding scaffolds with a potentially very large variety of bridging units and substituents on the terminal pyridine rings. The bridged 5-(2-chloropyridines) may also serve as precursors for the fabrication of metal ion-coordinated conjugated polymers. PMID- 10814086 TI - Steric strain and reactivity: electrophilic bromination of trans-(1-methyl-2 adamantylidene)-1-methyladamantane AB - trans-(1-Methyl-2-adamantylidene)-1-methyladamantane (DMAD, 1b) reacts with Br(2) in chlorinated hydrocarbon solvents to give either a bromonium polybromide ion pair or a substitution product, depending on bromine concentration. The first intermediate is a 1:1 pi-complex having K(f) = 1.85(0.19) x 10(3) M(-)(1) at 25 degrees C, which rapidly evolves to the bromonium tribromide ion pair. At high bromine concentration, which shifts all equilibria involving the counteranion of the ion pair intermediate toward the pentabromide species, this bromonium ion is stable and unable to further evolve into products. Temperature-dependent NMR spectra indicate chemical exchange of Br(+) between the sides of the plane containing the two carbons of the bromonium ion. At very low bromine concentration, no ionic intermediate is detected and the reaction rapidly yields a rearranged substitution product, identified as 10. Under these conditions the disappearance of the pi-complex follows a first-order rate law, and the observed rate constant increases with increasing olefin concentration, showing that product formation implies Br(-) as counteranion of the ionic intermediate, whose formation is a reversible process. A comparison of the results reported here for the bromination of 1b with those previously found for the parent olefin, adamantylideneadamantane (1a), shows that steric strain markedly affects the reactivity of the double bond. PMID- 10814087 TI - Spiropentane mimics of nucleosides: analogues of 2'-deoxyadenosine and 2' deoxyguanosine. Synthesis of all stereoisomers, isomeric assignment, and biological activity. AB - Synthesis of spirocyclic analogues of 2'-deoxyadenosine and 2'-deoxyguanosine (12a-15a and 12b-15b) is described. Rhodium-catalyzed reaction of ethyl diazoacetate with methylenecyclopropane 19, obtained from 2-bromo-2 bromomethylcyclopropane 17 via debromination (16), reduction (18), and acetylation (19), gave a mixture of all four isomeric spiropentanes 20a-20d. Hydrolysis afforded hydroxy carboxylic acids 21a-21d. Acetylation of separated proximal + medial-syn isomers 21a + 21b and medial anti + distal isomers 21c + 21d furnished acetates 22a + 22b and 22c + 22d. Curtius rearrangement effected by diphenylphosphoryl azide in tert-butyl alcohol performed separately with mixtures 22a + 22b and 22c + 22d led to BOC-amino spiropentanes 23a + 23b and 23c + 23d. After deacetylation all isomers 24a-24d were separated and deprotected to give aminospiropentane hydrochlorides 25a-25d. Free bases were of limited stability. The heterocyclic moieties were introduced into individual isomers 25a-25d via 6 chloropurine derivatives 26a-26d or 30a-30d. Ammonolysis of 26a-26d furnished the adenine isomeric series 12a-15a, whereas guanine derivatives 12b-15b were obtained by hydrolysis of 30a-30d with formic acid. The isomeric assignments followed from IR spectra of BOC-aminospiropentanes 24a-24d and NMR spectra of 12a 15a including NOE and (H,H) COSY. The proximal and medial-syn isomers 12a and 12b were modest inhibitors of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) in culture, whereas the medial-anti isomer 12c was a substrate for adenosine deaminase. The distal isomer 15b was an anti-EBV agent. The medial-syn phosphoralaninate 34 was an effective inhibitor of HCMV replication in vitro. It was also active against herpes simplex virus type 1 (HSV-1), varicella zoster virus (VZV), human immunodeficiency virus (HIV-1), hepatitis B virus (HBV), and EBV with a varying degree of cytotoxicity. PMID- 10814089 TI - Stereoselective synthesis of beta-benzyl-alpha-alkyl-beta-amino acids from L aspartic acid. AB - A stereoselective synthesis of beta-benzyl-alpha-alkyl-beta-amino acids 1 and 2 from L-aspartic acid 3 has been developed. Methyl 5-phenyloxazolidin-2-one-4 acetate 4 was prepared from L-aspartic acid 3 through the acylation of benzene or phenyllithium with alpha-amino carboxyl group of L-aspartic acid skeleton. Alkylation of a dianion of 4 with alkyl halides and subsequent hydrogenation afforded anti-disubstituted beta-amino acids 1b and 1c in high stereoselectivities. Complete reversal of the stereoselection was realized by the alkylation of 4-phenyl-3-tert-butoxycarbonylamino-4-butanolide 6 which was obtained in a single step from 4. The 2,3,4-trisubstituted amino lactone 7 thus obtained was hydrogenated to give a syn-disubstituted beta-amino acid 2a. The syn products 2b, 2c, and 2dwere alternatively prepared via aldol condensation of 6 with aromatic or aliphatic aldehydes followed by stereoselective reduction of the double bond with nickel chloride-sodium borohydride. PMID- 10814088 TI - Highly diastereoselective alpha-mannopyranosylation in the absence of participating protecting groups. AB - S-Phenyl 2,6-di-O-benzyl-3,4-O-(2',3'-dimethoxybutane-2', 3'-diyl)-1-thia-alpha-D mannopyranoside and its sulfoxide, following activation at -78 degrees C with benzenesulfenyl triflate or triflic anhydride, respectively, provide the corresponding alpha-mannosyl triflate as demonstrated by NMR spectroscopy. On addition of an acceptor alcohol alpha-mannosides are then formed. Similarly, S phenyl 2,3-O-carbonyl-4, 6-O-benzylidene-1-thia-alpha-D-mannopyranoside and ethyl 3-O-benzoyl-4, 6-O-benzylidene-2-O-(tert-butyldimethylsilyl)-1-thia-alpha-D mannopyr anoside both provide alpha-mannosides on activation with benzenesulfenyl triflate followed by addition of an alcohol. These results stand in direct contrast to the highly beta-selective couplings of comparable glycosylations with 2,3-di-O-benzyl-4, 6-O-benzylidene protected mannosyl donors and draw attention to the subtle interplay of reactivity and structure in carbohydrate chemistry. PMID- 10814091 TI - Supramolecular steric effects as the means of making reactive carbon radicals persistent. Quantitative characterization of the external surface of MFI zeolites through a persistent radical probe and a langmuir adsorption isotherm AB - The photochemistry of tetraphenylacetone (1) adsorbed on the external surface of a MFI zeolite (the sodium form of LZ-105) has been investigated in combination with computational chemistry, surface area measurements, EPR analysis, and classical adsorption isotherms. All of the methods are consistent with a supramolecular structural model in which 1 is first adsorbed strongly through intercalation of a single benzene ring into a hole on the LZ-105 external surface (site I) followed by a weaker binding to the external framework between the holes (site II) until a monolayer of 1 is formed. From both computational and surface area measurements, it is estimated that the site I holes on the external surface will be filled at ca. 0.3-0.5 wt %/wt loading of 1/LZ-105, which corresponds to 6.5 x 10(18) (ca. 10(-)(5) mol) of holes or molecules of 1 adsorbed in holes per gram of zeolite. The supramolecular composition of ca. 0.3-0.5% of 1 on LZ-105 characterizes a "break point" for the photochemistry and the EPR measurements, since it represents the value for saturation of the site I holes with 1. These conclusions are supported quantitatively by experimental isotherms of the adsorption of 1 on LZ-105. Photolysis of 1 intercalated in the site I holes causes fragmentation into two isomeric supramolecular diphenylmethyl (DPM) radicals, one (DMP)(in) which is adsorbed into the internal surface and becomes strongly persistent (half-life of many weeks) and the other (DMP)(ex) which diffuses on the external surface and rapidly dimerizes (less than a few minutes) to produce the radical-radical combination product tetraphenylethane (2). Photolysis of 1 adsorbed on the solid external surface produces two supramolecularly equivalent DPM radicals (DMP)(ex) that diffuse on the external surface and rapidly dimerize to produce 2, and do not produce persistent DPM radicals. PMID- 10814090 TI - Cyclopropane-derived peptidomimetics. Design, synthesis, and evaluation of novel enkephalin analogues. AB - It is known that peptide mimics containing trans-substituted cyclopropanes stabilize extended conformations of oligopeptides, and molecular modeling studies now suggest that the corresponding cis-cyclopropane dipeptide isosteres could stabilize a reverse turn. To begin to assess this possibility, a series of cis substituted cyclopropanes were incorporated as replacements of the Gly(2)-Gly(3) and Phe(4)-Leu(5) dipeptide subunits in Leu-enkephalin (H(2)N-Tyr-Gly-Gly-Phe-Leu OH), which is believed to bind to opiod receptors in a conformation containing a beta-turn. General methods for the synthesis of the cyclopropane-containing dipeptide isosteres -XaaPsi[COcpCO]Yaa- and -XaaPsi[NHcpNH]Yaa-were developed by a sequence that featured the enantioselective cyclization of allylic diazoacetates catalyzed by the chiral rhodium complexes Rh(2)[(5S)-MEPY](4) and Rh(2)[(5R)-MEPY](4). A useful modification of the Weinreb amidation procedure was applied to the opening of the intermediate lactones with dipeptides, and a novel method for the synthesis of substituted diaminocyclopropanes was also developed. The Leu-enkephalin analogues were tested in a panel of binding and functional assays, and although those derivatives containing cyclopropane replacements of the Gly(2)-Gly(3) exhibited low micromolar affinity for the mu-receptor, analogues containing such replacements for the Phe(4)-Leu(5) subunit did not bind with significant affinity to any of the opioid receptors. These results are discussed. PMID- 10814092 TI - Effects of AB - Effects of cetyltrimethylammonium bromide (CTABr) micelles on second-order rate constants (k(n)(obs)) for nucleophilic reactions of amines (piperidine and n butylamine) with ionized phenyl salicylate (PS(-)) reveal a nonlinear decrease with the increase in [D(n)] (where [D(n)] = [CTABr](T) - cmc) at a constant [NaBr] and 35 degrees C. The observed data, at a constant [NaBr], fit reasonably well to a pseudophase model of micelles, and such a data fit gives kinetic parameters such as CTABr micellar binding canstant (K(S)) of PS(-). The effect of [NaBr] upon K(S) is explained with the empirical relationship K(S) = K(S)(0)/(1 + psi[NaBr]), where psi is an empirical parameter. PMID- 10814093 TI - Asymmetric synthesis of axially chiral biaryls via desymmetrization of 2,2',6,6' tetrahydroxybiphenyl using 1,4-Di-O-benzyl-L-threitol as a chiral template AB - Sequential etherification of 2,2',6,6'-tetrahydroxybiphenyl (1) with 1,4-di-O benzyl-L-threitol under Mitsunobu conditions gives desymmetrized biphenyldiol 9 of S-axial chirality exclusively. Cyclization of 9 with 1,omega-dibromoalkanes followed by removal of the chiral auxiliary yields (S)-2,2'-biphenyldiols 14 with alkylenedioxy bridges at the 6 and 6' positions. (S)-6,6'-Dialkyl- and diphenyldiols 20 are obtained in an efficient manner via Pd(0)-catalyzed cross coupling of bis(triflate) derivative 17 with organozinc reagents. Bis(triflate) 17 also serves as an intermediate for asymmetric synthesis of axially chiral biphenyldicarboxylic acid 23, terphenylcarboxylic acid 28, lactone 26, and lactam 30. PMID- 10814094 TI - Effective ring-opening reaction of aziridines with trimethylsilyl compounds: A facile access to beta-amino acids and 1,2-diamine derivatives AB - Ring-opening reactions of aziridines with trimethylsilyl compounds triggered by tetrabutylammonium fluoride give the corresponding products regioselectively in excellent yield. It provides a facile and efficient procedure for the ring opening reactions of aziridines and affords a practical access to the synthesis of cyano-, azido-, or chloroamines because of its efficiency and simplicity. The products are easily transformed to vicinal diamines or beta-amino acids. PMID- 10814095 TI - Amphidinolide T, novel 19-membered macrolide from marine dinoflagellate Amphidinium sp. AB - A novel 19-membered macrolide, amphidinolide T (1), has been isolated from a marine dinoflagellate Amphidinium sp., and the structure was elucidated on the basis of spectroscopic data. Relative stereochemistry at C-7, C-8, and C-10 was deduced from the NOESY correlations, while absolute configurations at C-2, C-13, C-14, and C-18 were assigned on the basis of NMR data of the MTPA esters of 1 and those of degradation products of 1. PMID- 10814096 TI - Synthesis and chiroptical properties of Methanocycloocta AB - The synthesis of chiral methanocyclocta[b]indoles, the fused structures obtained from enantiomeric bicyclo[3.3.1]nonanones via Fisher indolization reaction, is reported. The starting optically active bicyclo[3.3.1]nonane-2,6-dione (1) was obtained by a chiral HPLC enantiomer separation on a swollen microcrystalline triacetylcellulose column and by the enzymatic resolution of the racemic dione. The circular dichroism (CD) spectra of the chiral structures 4, 5, and 7 were recorded, and the absolute configuration for the indole compounds was assigned. The theoretical calculations of the CD spectrum of diindole 4 reproduce the (1)B(b) couplet at 229 nm but predict wrong signs for the (1)L(a) and (1)L(b) bands using standard polarization directions. The CD spectrum of indole ketone 5 is reproduced correctly. PMID- 10814098 TI - Theoretical prediction of the stability and intramolecular rearrangement reactions of heteroanalogues of cyclopropylcarbene: 2-oxiranyl-, 2-aziridinyl-, and 1-aziridinylcarbene AB - Ab initio studies have been carried out on the experimentally uncharacterized 2 oxiranyl- (8), fluoro-2-oxiranyl- (15), 2-aziridinyl- (21), and 1 aziridinylcarbene (29) on the B3LYP/6-31G(d) and CCSD(T)/6-31G(d,p)//MP2/6 31G(d,p) levels of theory. Like the parent cyclopropylcarbene (1), the beta heterosubstituted carbenes 8 and 21-intermediates in the Eschenmoser-Tanabe fragmentation-are predicted to give either ethyne extrusion or ring expansion reactions, depending on the initial conformation. The barriers for these reactions are low (<6 kcal mol(-)(1)), while the interconversion of the carbene conformers requires higher energies. NBO analyses show that the donor-acceptor interactions between the carbene frontier orbitals and sigma-bond orbitals in the oxiranyl and aziridinyl ring destabilize the compounds kinetically. 2-Oxiranyl- and 2-aziridinylcarbenes with a fixed cis (=exo) conformation are predicted to rearrange to the heteroanalogue derivatives of cyclobutene. The fluoro substituted 2-oxiranylcarbene 15 is predicted to rearrange to the substituted oxete via one equilibrated conformer, due to the low barrier of the C-(CF) rotation. 1-Aziridinylcarbene 29 is the thermodynamically most stable of the investigated carbenes; however, the fragmentation into HCN and ethene is a facile reaction with a calculated barrier of 15 kcal mol(-)(1), which confirms experiments. PMID- 10814097 TI - Synthesis of indan-based unusual alpha-amino acid derivatives under phase transfer catalysis conditions. AB - Conformationally constrained cyclic alpha-amino acid derivatives were synthesized under solid-liquid phase-transfer catalysis conditions. This methodology involves the bis-alkylation of ethyl isocyanoacetate with various alpha,alpha'-dibromo-o xylene derivatives [alpha,alpha'-dibromo-o-xylene 5, 2,3-bis(bromomethyl)-1, 4 dimethoxybenzene 6, 1,2-bis(bromomethyl)-4,5-dibromobenzene 7, 2, 3 bis(bromomethyl)naphthalene 8, 1,8-bis(bromomethyl)-naphthalene 9, 6,7 bis(bromomethyl)-2,2-dimethyl-1H-phenalene-1,3(2H)-dione 10, 2, 3 bis(bromomethyl)-1,4-anthraquinone 11, 6, 7-bis(bromomethyl)quinoxaline 12, 3,4 bis(bromomethyl)furan 13, 1,2, 4,5-tetrakis(bromomethyl)benzene 28, and hexakis(bromomethyl)benzene 30] using potassium carbonate as a base and tetrabutylammonium hydrogensulfate as a phase-transfer catalyst to give corresponding isonitrile derivatives, which upon hydrolysis with HCl in ethanol gave amino esters. Using this method electron-deficient as well as electron-rich and halogen-substituted indan-based alpha-amino acids were prepared. The preparation of bis-indan as well as tris-indan alpha-amino esters is also described. PMID- 10814099 TI - Flash vacuum pyrolysis of methoxy-substituted lignin model compounds. AB - The flash vacuum pyrolysis (FVP) of methoxy-substituted beta-O-4 lignin model compounds has been studied at 500 degrees C to provide mechanistic insight into the primary reaction pathways that occur under conditions of fast pyrolysis. FVP of PhCH(2)CH(2)OPh (PPE), a model of the dominant beta-O-4 linkage in lignin, proceeds by C-O and C-C cleavage, in a 37:1 ratio, to produce styrene plus phenol as the dominant products and minor amounts of toluene, bibenzyl, and benzaldehyde. From the deuterium isotope effect in the FVP of PhCD(2)CH(2)OPh, it was shown that C-O cleavage occurs by homolysis and by 1,2-elimination in a ratio of 1.4:1, respectively. Methoxy substituents enhance the homolysis of the beta-O 4 linkage, relative to PPE, in o-CH(3)O-C(6)H(4)OCH(2)CH(2)Ph (o-CH(3)O-PPE) and (o-CH(3)O)(2)-C(6)H(3)OCH(2)CH(2)Ph ((o-CH(3)O)(2)-PPE) by a factor of 7.4 and 21, respectively. The methoxy-substituted phenoxy radicals undergo a complex series of reactions, which are dominated by 1,5-, 1,6-, and 1,4-intramolecular hydrogen abstraction, rearrangement, and beta-scission reactions. In the FVP of o CH(3)O-PPE, the dominant product, salicylaldehyde, forms from the methoxyphenoxy radical by a 1,5-hydrogen shift to form 2-hydroxyphenoxymethyl radical, 1,2 phenyl shift, and beta-scission of a hydrogen atom. The 2-hydroxyphenoxymethyl radical can also cleave to form formaldehyde and phenol in which the ratio of 1, 2-phenyl shift to beta-scission is ca. 4:1. In the FVP of o-CH(3)O-PPE and (o CH(3)O)(2)-PPE, products (ca. 20 mol %) are also formed by C-O homolysis of the methoxy group. The resulting phenoxy radicals undergo 1,5- and 1,6-hydrogen shifts in a ratio of ca. 2:1 to the aliphatic or benzylic carbon, respectively, of the phenethyl chain. In the FVP of (o-CH(3)O)(2)-PPE, o-cresol was the dominant product. It was formed by decomposition of 2-hydroxy-3 hydroxymethylbenzaldehyde and 2-hydroxybenzyl alcohol, which are formed from a complex series of reactions from the 2, 6-dimethoxyphenoxy radical. The key step in this reaction sequence was the rapid 1,5-hydrogen shift from 2-hydroxy-3 methoxybenzyloxy radical to 2-hydroxymethyl-6-methoxyphenoxy radical before beta scission of a hydrogen atom to give the substituted benzaldehyde. The 2 hydroxybenzyl alcohols rapidly decompose under the reaction conditions to o benzoquinone methide and pick up hydrogen from the reactor walls to form o cresol. PMID- 10814100 TI - Novel terpenoids from the West Indian sea whip Pseudopterogorgia elisabethae (Bayer). Elisapterosins A and B: rearranged diterpenes possessing an unprecedented cagelike framework. AB - Four diterpenes and a nor-diterpenoid, all of which possess unusual carbocyclic skeletons, were isolated from the hexane solubles of the West Indian gorgonian Pseudopterogorgia elisabethae. The structures and relative configurations of novel metabolites elisabethin D (2), elisabethin D acetate (3), 3-epi elisabanolide (5), elisapterosin A (6), and elisapterosin B (7) were elucidated by interpretation of overall spectral data, which included 2D NMR correlation methods, IR, UV, and accurate mass measurements (HREI-MS and HRFAB-MS), chemical reactions, and X-ray diffraction analyses. The tetracyclic carbon skeleton of the elisapterosins is undescribed and constitutes a new class of C(20) rearranged diterpenes. Elisapterosin B displays strong in vitro anti-tuberculosis activity. PMID- 10814101 TI - A practical and efficient preparation of the releasable naphthosultam side chain of a novel anti-MRSA carbapenem. AB - A practical large-scale synthesis of the naphthosultam-based side chain of the anti-MRSA antibiotic 1 has been achieved in 29% overall yield over seven steps from 1-methylnaphthalene. The synthesis was completed without the use of protecting groups, featuring a novel naphthosultam annelation, a chemoselective acid-catalyzed triflation, and the use of a novel naphthosultam dianion to effect functionalization through benzylic metalation. PMID- 10814102 TI - Spontaneous hydrolysis reactions of cis- and trans-beta-methyl-4-methoxystyrene oxides (Anethole oxides): buildup of trans-anethole oxide as an intermediate in the spontaneous reaction of cis-anethole oxide. AB - Rates and products of the reactions of trans- and cis-beta-methyl-4 methoxystyrene oxides (1 and 2) (anethole oxides) and beta,beta-dimethyl-4 methoxystyrene oxide (3) in water solutions in the pH range 4-12 have been determined. In the pH range ca. 8-12, each of these epoxides reacts by a spontaneous reaction. The spontaneous reaction of trans-anethole oxide (1) yields ca. 40% of (4-methoxyphenyl)acetone and 60% of 1-(4-methoxyphenyl)-1, 2 propanediols (erythro:threo ratio ca. 3:1). The spontaneous reaction of cis anethole oxide is more complicated. The yields of diol and ketone products vary with pH in the pH range 8-11, even though there is not a corresponding change in rate. These results are interpreted by a mechanism in which 2 undergoes isomerization in part to the more reactive trans-anethole oxide (1), which subsequently reacts by acid-catalyzed and/or spontaneous reactions, depending on the pH, to yield diol and ketone products. The buildup of the intermediate trans anethole oxide in the spontaneous reaction of cis-anethole oxide was detected by (1)H NMR analysis of the reaction mixture. Other primary products of the spontaneous reaction of 2 are (4-methoxyphenyl)acetone (73%) and threo-1-(4 methoxyphenyl)-1,2-propanediol (ca. 3%). The rates and products of the spontaneous reaction of 2 and its beta-deuterium-labeled derivative were determined, and the lack of significant kinetic and partitioning deuterium isotope effects indicates that the isomerization of 2 to ketone and to trans anethole oxide must occur primarily by nonintersecting reaction pathways. PMID- 10814104 TI - Preparation, X-ray crystal structures, and reactivity of alkynylcyclopropenylium salts AB - Several 1,2-diphenyl-3-alkynylcyclopropenes were prepared by the reaction of acetylenic nucleophiles with diphenylcyclopropenylium perchlorate. The cyclopropenes were converted into alkynylcyclopropenylium salts via hydride abstraction with triphenylmethyl cation. Attempts to prepare a dication from either ethyne-bridged or butadiyne-bridged biscyclopropenes produced only the corresponding monocations. A dication was prepared when an ethylene spacer was inserted between the acetylene groups of the butadiyne-bridged biscyclopropene. Single-crystal X-ray structures of three of the cyclopropenylium ions were obtained. pK(R+) titrations were carried out on two of the salts, which showed that acetylene substituents provide about the same degree of stability to the cyclopropenylium core as a phenyl group. Nucleophilic addition to the alkynylcyclopropenylium ions under kinetic conditions gave a statistical mixture of products; however, under thermodynamic reaction conditions, nucleophilic addition followed by ring opening produced only one of three possible products. Calculations at the ab initio level were carried out to determine the charge distribution of the cations. PMID- 10814103 TI - Synthesis of the repeating decapeptide unit of Mefp1 in orthogonally protected form. AB - Mefp1 is a protein produced by the marine mussel, Mytilus edulis, which helps the organism to adhere to surfaces in turbulent waters. To better understand the nature of the adhesion process, we sought to synthesize homogeneous oligopeptides based on the repeating decapeptide unit of the protein. The fully protected decapeptide 10 has been synthesized from appropriately protected amino acid building blocks using a fragment condensation strategy. A key feature of the strategy is the late incorporation of the synthetically valuable dihydroxyproline residue. This synthesis of the orthogonally protected repeating decapeptide unit of Mefp1 represents an important first step toward producing useful quantities of homogeneous oligopeptides related to the protein. PMID- 10814105 TI - Preparation of optically active allylic hydroperoxy alcohols and 1, 3-diols by enzyme-catalyzed kinetic resolution and photooxygenation of chiral homoallylic alcohols. AB - All four possible enantiomers of the 3-hydroperoxy-4-penten-1-ols 2a, b and their corresponding 4-pentene-1,3-diols 4a,b have been prepared for the first time in high enantiomeric purity (up to 98% ee) and in preparative amounts according to two distinct ways: First the photooxygenation of the racemic homoallylic alcohols 1 gave the racemic hydroperoxy alcohols 2, which have subsequently been kinetically resolved by horseradish peroxidase (HRP); alternatively, first the lipase-catalyzed resolution afforded the optically active homoallylic alcohols 1 and subsequent photooxygenation led to the enantiomerically enriched hydroperoxy alcohols 2. PMID- 10814106 TI - Triphenylsilyl as a protecting group in the synthesis of 1, 12 heterodisubstituted p-carboranes AB - A triphenylsilyl group is used as an auxiliary in the synthesis of heterodisubstituted p-carboranes via triphenylsilyl-p-carborane (1). The preparation of 1 is statistical, but with recovery of the starting p-carborane, the effective conversion to 1 is about 90%. Carborane 1 has been easily converted to its lithium and copper derivatives, which were reacted with a range of electrophiles including alkyl halides: an aryl iodide, an acetylene bromide, and a sulfenyl chloride. The derivatives of 1 are crystalline and UV active, which facilitates their isolation and purification. The Ph(3)Si group is efficiently removed with fluoride to give monosubstituted p-carboranes I, which upon further nucleophilic substitution yield p-carboranes II. The yields of heterodisubstituted products II are higher than for direct, "statistical" syntheses. For example, 12-pentyl-1, 12-dicarba-closo-dodecaborane-1-carboxylic acid was synthesized from p-carborane via 1 in 62% overall yield, a considerable improvement over the 44% yield obtained by direct methods. PMID- 10814107 TI - Synthesis of N-(Hydroxy)amide- and N-(Hydroxy)thioamide-containing peptides. AB - Methods developed with N-(benzoyloxy)amines and hydroxamic acids were used in the synthesis of N-(hydroxy)amide-containing pseudopeptides. Acylation of N (benzoyloxy)phenethylamine with the acid chloride of N(alpha)-Fmoc-L-leucine provided a N(alpha)-Fmoc-N-(benzoyloxy)-L-leucinamide in 90% yield. Deprotection of the benzoyl group (using 10 vol % NH(4)OH/MeOH) provided the N(alpha)-Fmoc-N (hydroxy)-L-leucinamide in 87% yield. In general, the appended Fmoc group allowed for further elaboration of the N-hydroxy-N-(alkyl)amides using classic peptide coupling methods. A practical synthetic strategy was developed, and racemization issues were addressed using diastereomeric Val-Leu derivatives. In addition, N (hydroxy)thioamides were generated from the corresponding N (benzoyloxy)thioamides. N-(Benzoyloxy)thioamides were obtained in moderate yields (53-76%) from the reaction of the corresponding N-(benzoyloxy)amides with Lawesson's reagent (i.e., 2, 4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane 2,4-disu lfide). In summary, this new technology allows for the introduction of either N-hydroxyamide or N-(hydroxy)thioamide linkages into pseudopeptide chains without racemization. PMID- 10814108 TI - n-sigma charge-transfer interaction and molecular and electronic structural properties in the hydrogen-bonding systems consisting of p-quinone dianions and methyl alcohol. AB - Molecular and electronic structural properties of the hydrogen-bonded complexes of p-quinone dianions (PQ(2)(-)) were investigated by electrochemistry and spectroelectrochemistry of PQ in MeCN combined with ab initio MO calculations. Hydrogen bonding between PQ(2)(-) and MeOH was measured as the continuous positive shift of the apparent second half-wave reduction potentials with increasing concentrations of MeOH. Detailed analyses of the behavior reveal that PQ(2)(-) forms the 1:2 hydrogen-bonded complexes at low concentrations of MeOH and the 1:4 complexes at high concentrations, yielding the formation constants. Temperature dependence of the formation constants allows us to yield the formation energy as 76.6 and 118.9 kJ mol(-)(1) for the 1:2 and 1:4 complex formation of the 1,4-benzoquinone dianion (BQ(2)(-)) with MeOH, respectively. These results show that the pi-dianions involving the quinone carbonyl groups exhibit very strong hydrogen-accepting ability. The longest wavelength band of the spectra of BQ(2)(-) and the chloranil dianion (CL(2)(-)) is assigned to the (1)B(3u) <-- (1)A(g) band mainly contributed from an intramolecular charge transfer (CT) configuration. Hydrogen bonding allows the band of BQ(2)(-) and CL(2)(-) to be blue-shifted, depending on the strength of the hydrogen bonds. CNDO/S-CI calculations reveal that the blue shift is ascribed to stabilization of the ground state by the hydrogen bonding involving strong n-sigma-type CT interaction. The HF/6-31G(d) calculation results show that the structure of PQ(2)(-) is characterized by a lengthening of the C=O bonds and a benzenoid ring. The geometrical properties of the hydrogen-bonded complexes of PQ(2)(-) are a slight lengthening of the C=O bonds and a short distance of the hydrogen bonds. It is demonstrated that this situation is due to the strong n-sigma CT interaction in the hydrogen bonds. The results suggest that the differing functions and properties of biological quinones are conferred by the n-sigma CT interaction through hydrogen bonding of the dianions with their protein environment. PMID- 10814109 TI - Preparation of trifluoromethyl aryl sulfides using Silver(I) trifluoromethanethiolate and an inorganic iodide AB - Reaction of silver(I) trifluoromethanethiolate (AgSCF(3)) with KI or tetra-n butylammonium iodide in acetonitrile leads to the formation of a nucleophilic source of trifluoromethanethiolate. This source is capable of converting activated fluoro-, chloro-, bromo-, and iodoaromatics into the corresponding trifluoromethyl aryl sulfides under mild conditions. After successful reaction with tetra-n-butylammonium iodide, crystals of Bu(4)N[Ag(3)I(4)] precipitate from the reaction mixture. With less activated aromatic compounds, decomposition of the trifluoromethanethiolate anion occurs preferentially, giving bis(trifluoromethyl)disulfide, tetrakis(trifluoromethylthio)ethene, and 3,4,5, 6 tetrakis(trifluoromethylthio)-1,2-dithiine. The use of copper(I) trifluoromethanethiolate and mercury(II) trifluoromethanethiolate for such reactions has also been investigated. PMID- 10814110 TI - Structural and solvent effects on the mechanism of base-induced rearrangement of epoxides to allylic alcohols AB - A combined experimental and computational study is presented which explores the influence of structure and solvent on the base-catalyzed isomerization of cyclopentene- and cyclohexene oxides. Cyclohexene oxide is known to rearrange via a syn beta-elimination in nonpolar solvents. Cyclopentene oxide instead undergoes alpha-elimination to a carbenoid intermediate in nonpolar solvents due to the unusual acidity of the alpha-proton, not because of an unfavorable conformation. In HMPA, cyclopentene oxide undergoes beta-elimination. To explore the origins of this mechanistic change, deuterium-labeled cis-4-tert-butylcyclohexene oxide was rearranged in HMPA and was found to react via anti beta-elimination, as presumably do cyclopentene oxide and other epoxides. PMID- 10814112 TI - Direct carbonylation at a C-H bond in the benzene ring of 2-phenyloxazolines catalyzed by Ru(3)(CO)(12). Scope, limitations, and mechanistic aspects AB - The ruthenium-catalyzed carbonylation at a C-H bond in the benzene ring of a 2 phenyloxazoline is described. The reaction of 2-phenyloxazolines with CO and ethylene in toluene in the presence of a catalytic amount of Ru(3)(CO)(12) resulted in propionylation at an ortho C-H bond in the benzene ring. The presence of the oxazoline ring on the benzene ring is essential for the carbonylation to proceed. Other heterocycles, such as oxazine, oxazole, and thiazoline rings, also served as acceptable directing groups as did the oxazoline ring. A wide functional group compatibility was observed. The site selectivity of the carbonylation was examined using meta-substituted phenyloxazolines. It was found that the carbonylation took place exclusively at the less-hindered C-H bond, irrespective of the nature of substituents, indicating that the site selectivity was determined by steric factors. The reaction was also applicable, not only to a benzene ring, but also to naphthyl and thiophenyl rings. Olefins such as propene and trimethylvinylsilane in place of ethylene could also be used in the carbonylation reaction, while other olefins, such as 1-hexene, tert butylethylene, vinylcyclohexane, isoprene, 1,5-hexadiene, cyclohexene, 1, 5 cyclooctadiene, styrene, methyl acrylate, vinyl acetate, allyltrimethylsilane, and triethoxyvinylsilane did not afford the coupling products. An equilibrium between 2-phenyloxazolines, carbon monoxide, and olefins exists on one hand and the corresponding ketones on the other hand, and product composition is governed by the equilibrium thermodynamics of the system. The results of deuterium labeling experiments suggest that the catalysis involves a reversible C-H bond cleavage and that the rate-determining step is not the cleavage of a C-H bond. The results of kinetic study of the effects of CO pressure show that the reaction rate accelerates with decreasing CO pressure. PMID- 10814111 TI - Two comparisons of the performance of positional scanning and deletion synthesis for the identification of active constituents in mixture combinatorial libraries. AB - Two libraries of 120 compounds each were prepared as individual compounds and as full mixtures. The corresponding scanning and deletion synthesis deconvolution libraries were prepared and tested (L-1210, IC(50)) alongside the individual compounds and mixture libraries. This testing, where the properties of each compound in the mixtures were known, was used to compare the performance of scanning and deletion deconvolution libraries. Each has its own intrinsic strengths, with the former being capable of identifying multiple hits at the expense of accurately identifying the most potent library member, while the latter typically is more sensitive to identifying the most potent hit but at the expense of differentiating weaker activities. The protocols complement one another and together more thoroughly identify potent library members. PMID- 10814113 TI - Synthesis and study of C(3)-symmetric hydropyran cyclooligolides with oriented aryl and alcohol appendages at 10 A spacing. AB - Modular syntheses of C(3)-symmetric macrocycles with pendant aryl and hydroxymethyl groups are described. These functional groups, amenable to further elaboration, were installed early in each synthesis and carried through an iterative sequence of module coupling and macrolactonization. Association constants for macrolides 1a-c with alkali metal cation guests were determined, and sandwich-type complexes with Ba(2+) were confirmed for these macrocycles based on (1)H NMR studies, including Job plots. X-ray crystallographic data for macrolides 1a and 1c were obtained and are discussed in detail. These data provide support that the macrolides are structurally well-defined and preorganized for binding the potassium cation. Preparation of the tris(bromoacetylated) macrotriolide 43 exemplifies a functionalized platform suitable for elaboration with peptide or carbohydrate residues. PMID- 10814115 TI - Oxovanadium(V)-induced cross-coupling reaction between two ligands of organozinc compounds AB - Oxovanadium(V) compounds such as VO(OEt)Cl(2) serve as useful oxidants for organozinc compounds, providing the corresponding cross-coupling products derived from two ligands of organozinc compounds. In particular, triorganozincates undergo selective cross-coupling smoothly by the action of oxovanadium(V). PMID- 10814114 TI - Stereoselective synthesis of C5-C20 and C21-C34 subunits of the core structure of the aplyronines. Applications of enantioselective additions of chiral allenylindium reagents to chiral aldehydes. AB - The stereoselective synthesis of a C5-C20 and a C21-C34 subunit of the aplyronine family of polyketide marine macrolides has been achieved. These subunits contain all 15 stereocenters of the core structure. Six of the 15 stereocenters were introduced through enantioselective and diastereoselective additions of chiral allenylindium reagents to alpha-methyl-beta-oxygenated propionaldehydes. The products of these additions were further transformed by reactions involving the terminal alkynyl substituent produced in the addition reactions. Unlike previous applications of this methodology, the present synthesis employs Pd(0)-catalyzed transmetalations of chiral allenylpalladium intermediates to generate the chiral allenylindium reagents in situ. PMID- 10814116 TI - An investigation of imidazole and oxazole syntheses using aryl-substituted TosMIC reagents. AB - This article describes efficient and mild protocols for preparing polysubstituted imidazoles in a single pot from aryl-substituted tosylmethyl isocyanide (TosMIC) reagents and imines generated in situ. Traditional imine-forming reactions employing virtually any aldehyde and amine followed by addition of the TosMIC reagent delivers 1,4,5-trisubstituted imidazoles with predictable regiochemistry. Employing chiral amines and aldehydes, particularly those derived from alpha amino acids, affords imidazoles with asymmetric centers appended to N-1 or C-5 with excellent retention of chiral purity. 1,4-Disubstituted imidazoles are also readily prepared by a simple variant of the above procedure. Selecting glyoxylic acid as the aldehyde component of this procedure leads to intermediates such as 48, which readily undergo decarboxylation and elimination of the tosyl moiety to deliver 1,4-disubstituted imidazoles in high yields. Alternatively, using NH(4)OH as the amine component in conjunction with a variety of aldehydes delivers 4, 5 disubstituted imidazoles in moderate to good yields in a single pot while avoiding the need for protecting groups. Finally, the facile preparation of mono- and disubstituted oxazoles from these TosMIC reagents and aldehydes is described. PMID- 10814117 TI - Palladium-catalyzed heteroannulation of 1,3-dienes to form alpha-alkylidene-gamma butyrolactones. AB - alpha-Alkylidene-gamma-butyrolactones are readily prepared by the palladium catalyzed heteroannulation of a variety of 1,3-dienes by alpha-iodo and alpha bromo acrylic acids. The best results are obtained by employing a catalytic amount of the sterically hindered chelating alkyl phosphine D-t-BPF [(di-tert butylphosphino)ferrocene]. In most cases, this process is highly regioselective. The reaction is believed to proceed via (1) oxidative addition of the vinylic halide to Pd(0), (2) organopalladium addition to the less hindered end of the 1,3 diene to form a pi-allylpalladium intermediate, and (3) nucleophilic displacement of the palladium by the carboxylate ion. PMID- 10814118 TI - Porphyrins with exocyclic rings. 14. Synthesis of tetraacenaphthoporphyrins, a new family of highly conjugated porphyrins with record-breaking long-wavelength electronic absorptions. AB - The Soret band for porphyrins is usually observed in the near-ultraviolet at approximately 400 nm, and few examples of "nonexpanded" porphyrins with this major absorption band at values above 500 nm have previously been reported in the literature. Ring fusion with aromatic ring systems such as naphthalene, anthracene, or phenanthrene generally only produces minor bathochromic shifts to this diagnostic absorption band. In this paper, the synthesis of a series of tetraacenaphthoporphyrins and their metal chelates is reported. The compact nature of the acenaphthylene ring system allows the introduction of meso substituents using the Lindsey methodology. meso-Tetraphenylporphyrin 10a shows the presence of a Soret band at 556 nm, while p-methoxy and p-nitro substituents in 10f and 10g, respectively, further shift this band to 560 and 570 nm. Addition of TFA produces the corresponding dications with slightly higher wavelength Soret bands at 565, 573, and 588 nm. These values compare to 525 nm for the dication of tetraacenaphthylene 8, which lacks the meso-aryl substituents, indicating that steric crowding and its resulting distortion of the macrocyclic conformation is responsible for a significant albeit minor portion of these shifts. The nickel(II), copper(II), and zinc chelates of 10a produce Soret bands at 528, 545, and 558 nm, respectively, demonstrating that the trend for increasing red shifts in metalloporphyrins across the periodic table is retained for this series. The lead(II) chelate 19d gave an additional "hyper" shift that brought the Soret band to 604 nm. A similar red shift could be achieved by introducing four phenylethynyl substituents at the meso positions, and this highly conjugated porphyrin (20) also showed a Soret band at 604 nm, while the corresponding dication afforded this absorption band at 629 nm. The essentially additive "hyper" shift due to lead chelation brought the Soret band for the related lead(II) complex 22d to 642 nm. These effects are by far the largest ever observed for true porphyrins and demonstrate that the Soret band can be fined tuned to virtually any part of the visible spectrum. PMID- 10814120 TI - A theoretical study of curing reactions of maleimide resins through michael additions of amines PMID- 10814119 TI - A new rhodium catalyst: formation of PMID- 10814121 TI - A new method for synthesis of unsymmetrical ureas using electrochemically prepared trifluoroethyl carbamates. PMID- 10814123 TI - A novel regio- and stereoselective formal cross-coupling reaction of unsaturated silanes with arenediazonium tetrafluoroborates PMID- 10814124 TI - Stereodivergent synthesis of (E)- and (Z)-2-alken-4-yn-1-ols from 2-propynoic acid: A practical route via 2-alken-4-ynoates PMID- 10814122 TI - Barrier to rotation about the phenyl-carbonyl carbon bond of methyl benzoate by dynamic NMR spectroscopy and ab initio molecular orbital calculations. PMID- 10814126 TI - A versatile one-Pot synthesis of 1,3-substituted guanidines from carbamoyl isothiocyanates PMID- 10814125 TI - A new method for the synthesis of tri-tert-butyl diethylenetriaminepentaacetic acid and its derivatives. PMID- 10814127 TI - Palladium-catalyzed asymmetric allylic alkylation of barbituric acid derivatives: enantioselective syntheses of cyclopentobarbital and pentobarbital. PMID- 10814128 TI - A versatile approach to the synthesis of (+)-mannostatin A analogues. PMID- 10814129 TI - Biomimetic Fe(II)-mediated degradation of arteflene (Ro-42-1611). The first EPR spin-trapping evidence for the previously postulated secondary carbon-centered cyclohexyl radical. PMID- 10814131 TI - Structure and temperature effects on the cyclization of rigid bisacetylenes PMID- 10814130 TI - A facile access to 2-methylthio/alkoxy/amino-3-acylimidazo[1, 2-a]pyridines based on cupric chloride promoted oxidative ring closure of alpha-oxoketene N,S-, N,O-, and N,N-acetals. PMID- 10814133 TI - Asymmetric catalysis of the friedel-crafts reaction with fluoral by chiral binaphthol-derived titanium catalysts through asymmetric activation PMID- 10814132 TI - 1,3-dipolar cycloaddition of furfuryl nitrones with acrylates. A convenient approach to protected 4-hydroxypyroglutamic acids. PMID- 10814134 TI - Porphyrin clips derived from diphenylglycoluril. Synthesis, conformational analysis, and binding properties PMID- 10814135 TI - Stereoselective Synthesis of (Z)- and (E)-Allylic Silanes by Copper-Mediated Substitution Reactions of Allylic Carbamates with Grignard Reagents. AB - Both (Z)- and (E)-allylic silanes were prepared with high stereoselectivity by the copper-mediated substitution of allylic carbamates by organometallic reagents. The reaction of alkylmagnesium reagents with (E)-allylic carbamates provides (Z)-allylic silanes, whereas both alkylmagnesium and alkyllithium reagents react with (Z)-allylic carbamates to afford (E)-allylic silanes. Because Grignard reagents are often more facile to prepare than alkyllithium species, these reagents are the optimal nucleophiles for the synthesis of both (Z)- and (E)-allylic silanes. This method also allows readily available nonracemic allylic carbamates to be converted to chiral, nonracemic (Z)- and (E)-allylic silanes with high stereoselectivity. PMID- 10814136 TI - Thermal and Photochemical Isomerization of Tetraaryl Tetrakis(trifluoromethyl) AB - The isomerization of tetraaryl tetrakis(trifluoromethyl)[4]radialenes was studied. When type II (all-Z) isomers of 5,6,7,8-tetraaryl-5,6,7,8 tetrakis(trifluoromethyl)[4]radialenes were heated in tetralin at 170-200 degrees C, isomerization occurred to give mixtures of four [4]radialenes in a ratio of ca. I:II:III:IV = 1:10:5:1. However, when the isomeric mixtures were heated in the solid state at the same temperature, selective isomerization took place to give type II isomers in good selectivity (>91%). Upon irradiation with light, the type II isomers first isomerized to mixtures of the four [4]radialene isomers (I:II:III:IV = 2:2:48:48) and then rearranged to cyclobuta[b]naphthalenes via a 6pi-electrocyclic reaction followed by 1,3-hydrogen migration. PMID- 10814137 TI - New Insights into the Chemistry of Lithium Carbamoyls: Characterization of an Adduct (R(2)NC(O)CLi(OLi)NR(2)). AB - Studies of the reaction of lithium dicyclohexylamide with N,N-dibutylformamide, 1 formylpiperidine, and 4-formylmorpholine indicate that the equilibria resulting from these compounds are shifted toward the formation of an adduct, which quickly collapses to dicyclohexylamine and the lithiated carbamoyl anion derived from the initial disubstituted formamide. Further reactions of the lithium carbamoyl lead to a new adduct where a lithiated carbon is bounded to N, O, and a carbonyl functionality. The (13)C NMR analysis of the reaction mixtures showed the presence of similar intermediates in all cases: adducts of this type have not been reported before. These dilithiated intermediates were trapped with methyl iodide giving the corresponding doubly methylated derivatives. Isolation of substituted glyoxylamides and quantitative determination of the products yields constitute further evidence of the whole reaction scheme proposed. PMID- 10814138 TI - On Five- vs Six-membered Diacetal Formation from Threitol and the Intermediacy of Unusually Stable Protonated Species(1). AB - The long known, but hitherto poorly understood, thermodynamically controlled diacetalation of rac-threitol with alkylaldehydes provided bicyclic, cis tetraoxadecalin (TOD) ("66") and bi(dioxolanyl) (BDO) ("55") products, shown to be formed in acid-concentration and temperature-dependent ratio. The configurational and conformational isomeric diacetals obtained in four such reactions of substituted aldehydes (RCHO, R = CH(3), CH(2)Cl, CH(2)Br, CO(2)CH(3)) with rac-threitol were isolated and characterized. A variable acid concentration analysis of the equilibrium mixture of products in one such case (R = CH(2)Br) was performed and provided equilibrium constants and, hence, free energy differences among these products and their relatively stable protonated intermediates. The latter were rationalized by the unusually high proton-affinity calculated for the cis-TOD ("66") form. PMID- 10814139 TI - Electroactive, Internal Anthraquinonoid Dendritic Cores(1). AB - The modification of the extended 1 --> 3 anthraquinonoid monomer 1 by substitution of the central aliphatic moiety with a more rigid, aromatic moiety has allowed the syntheses of the new branched building block 6 possessing multiple, internal redox sites. This monomer was used to generate the first tier, dendritic macromolecules 8 and 9, which incorporate different internal redox centers within the infrastructure as well as the traditional peripheral multifunctionality. Using cyclic voltammetry experiments, the electrochemical response and electronic interactions between the two electroactive sites (the anthraquinonoid and nitroaromatic moieties) of these dendritic constructs were explored. PMID- 10814140 TI - Solvolysis of 2-Bicyclo AB - The solvolysis rate of 2-bicyclo[3.2.2]nonyl p-toluenesulfonate (6-OTs) was nearly equal to that of cycloheptyl p-toluenesulfonate in 2,2,2-trifluoroethanol (TFE). This indicates that the ethylene bridge in 6-OTs does not significantly enhance the rate and that 6-OTs ionizes without anchimeric assistance. The solvolysis of [1-(13)C]-2-bicyclo[3.2.2]nonyl p-toluenesulfonate in methanol or TFE gave 2-substituted bicyclo[3.2.2]nonane, exo-2-substituted bicyclo[3.3.1]nonane, 2-bicyclo[3.2.2]nonene (10), and 2-bicyclo[3.3.1]nonene (11), whose distributions of (13)C labels were determined by quantitative (13)C NMR analysis using a relaxation reagent. The (13)C labels were exclusively placed at only two positions, the ratios of them were not unity, and the labels in 10 were less extensively scrambled than those in other products. These results indicate that the 2-bicyclo[3.2.2]nonyl cation is classical and that 10 is formed at a former ionization stage than 2-substituted bicyclo[3.2.2]nonane. The (13)C redistributions for both exo-2-substituted bicyclo[3.3.1]nonane and 11, which are yielded via 1,3-hydride shift, were similar to that of 2-substituted bicyclo[3.2.2]nonane, suggesting that 1,3-hydride shift occurs mainly at the solvent-separated ion pair. PMID- 10814141 TI - Role of Heteroatoms in Diastereofacial Control in Cycloaddition to a Dissymmetric Cyclohexa-1,3-diene Moiety in a Polycyclic Framework. Remarkable Stereodirecting Influence of Distal Protective Groups. AB - Diels-Alder cycloaddition to several derivatives of a facially dissymmetric diene, the hexacyclo[7.5.1.0.(1,6)0.(6,13)0.(8,12)0(10,14)]pentadeca-2,4-diene 7,15-dione 4a, with a variety of dienophiles such as singlet oxygen, N phenyltriazolinedione, dimethyl acetylenedicarboxylate, maleic anhydride, and N methylmaleimide has been studied. The stereochemistry of the resulting adducts has been unambiguously secured by (1)H and (13)C NMR spectral data, chemical correlations, and X-ray crystal structure determination. While a variety of dienophiles undergo [4 + 2]-cycloadditions with 4a predominantly from the carbonyl face, protection of the carbonyl groups in 4a as simple mono- or bis acetals 4b-e or thioacetals 9a,b leads to complete reversal in selectivity, favoring addition from the cyclobutane face, with heterodienophiles and acetylenic dienophiles. The reversal in selectivity observed in mono- and bis acetals 4b-e has been attributed to unfavorable electrostatic interaction between the oxygen atom and the incoming dienophile. Whereas, in the case of thioacetals 9a,b, apart from unfavorable electrostatic interactions, Cieplak-type hyperconjugative interactions have to be given due consideration in order to account for the observed selectivities. Our studies highlight the role of simple protective groups (acetals in the present case) in modulating diastereoselection during [4 + 2]-cycloadditions. PMID- 10814142 TI - Catalysis of a Diels-Alder Reaction by Amidinium Ions. AB - Amidines and guanidines are important functional groups in molecular recognition and host-guest chemistry. Here it is shown that lipophilic amidinium ions catalyze a cycloaddition reaction representing the key step of the Quinkert-Dane estrone synthesis. Hydrogen-bond-mediated association with the organic cation leads to an electrophilic activation of the dienophile and to enhanced rates of the Diels-Alder reaction. The observed effects are similar to those expected from mild Lewis acids. In competition experiments, amidinium catalysis favors the reaction of the less electron deficient dienophile. PMID- 10814143 TI - Asymmetric Samarium-Reformatsky Reaction of Chiral alpha-Bromoacetyl-2 oxazolidinones with Aldehydes. AB - The samarium(II) iodide mediated asymmetric Reformatsky-type reaction of chiral 3 bromoacetyl-2-oxazolidinones with various aldehydes was studied. A series of chiral 4-substituted 2-oxazolidinones 1-3 and 5,5-disubstituted "SuperQuat" oxazolidinones 4-5 were employed as chiral auxiliaries of the alpha-bromoacetic acid. The reaction of 1 with various aldehydes gave the alpha-unbranched beta hydroxy carboximides in good yields with high diastereomeric excess values (up to >99% de). The majority of the reaction product derived from 5,5-diphenyl SuperQuat 5 were highly crystallinity; a single recrystallization yielding a diastereomerically pure product with the other diastereomer not detectable by spectroscopic methods. The absolute configurations of the beta-hydroxy carboximides were determined by signs of optical rotations of the corresponding known ethyl esters referring to the literature values. Hydrolytic cleavage of the appended of beta-hydroxy moieties from the auxiliary SuperQuats was readily achieved under mild conditions using lithium hydroxide; the corresponding carboxylic acids and the returned SuperQuats were obtained in good yields without any evidence of racemization. The first step of the reaction is the reduction of the alpha-bromo group to produce the samarium enolate, which adds to an aldehyde. The absolute configuration of the adduct (7i) derive from benzaldehyde was found to be R, with the samarium enolate favoring the transition state predicted from chelation control of the reagent; this is in analogy to the discussion that has been used for the corresponding titanium enolate. The stereochemistry of the reaction may be explained by incorporating the Nerz-Stormes-Thornton chair transition structure model. PMID- 10814144 TI - Planar Chirality: Synthesis and Transannular Reactions of Unsaturated Optically Active Azoninones Bearing E-Olefins. AB - The zwitterionic aza-Claisen rearrangement of optically active trans 4-silyloxy-2 vinylpyrrolidines and carboxylic acid fluoride generated nine-membered ring lactams with high yields. The reaction proceeded with an almost complete 1,4 chirality transfer and the exclusive generation of the E-double bond in the medium sized rings to cause additional planar chiral information. The initially formed azoninones were characterized by a pS-arrangement of the olefin with respect to the ring. The rather kinetically stable conformation underwent a flipping of the double bond to give the pR-azoninones as the thermodynamically stable products. The planar diastereomers were subjected to regio- and diastereoselective transannular ring contractions to give indolizidinones. The stereochemical outcome was strongly dependent from the planar chiral information of the double bond and the lactam unit. The so-formed optically active bicycles bearing a defined substitution pattern should serve as versatile building blocks in alkaloid synthesis. PMID- 10814145 TI - Preparation and Reactivity of 1,3-Bis(alkylthio)allenes and Tetrathiacyclic Bisallenes. AB - Reactions of Ph(2)C(3) dianion, prepared from 1,3-diphenylpropyne and n butyllithium, with alkyl thiocyanates or alkane dithiocyanates gave 1,3 bis(alkylthio)allenes 1 or tetrathiacyclic bisallenes 2, respectively. Thermal reactions of 1 gave thiophenes 4 and 7, benzothiepin 5, 1,2 bis(benzylidene)cyclobutane 6, thiete 8, and alpha,beta-unsaturated ketone 9, and the reactions of tetrathiacyclic bisallenes 2a gave a cyclic dimer, 1,2 bis(benzylidene)cyclobutane derivative 10, quantitatively. Irradiation of 1,3 bis(alkylthio)allenes 1 and tetrathiacyclic bisallenes 2a caused rearrangement to give alkynes 18, 20, and 21. In the irradiation of the cyclic bisallenes 2a, isomerizations from dl to meso and meso to dl isomers were also found. In the reactions of allenes 1 and cyclic bisallenes 2a with diphenyl diazomethane, the diazomethane reacted selectively with the double bond rather than with the sulfur atom. PMID- 10814146 TI - Conformational Studies by Dynamic NMR. 71.(1) Stereodynamics of Triisopropyl(aryl)silanes in Solution and in the Solid State. AB - A study has been carried out of the conformations of triisopropyl(aryl)silanes (i Pr)(3)SiAr, (Ar = phenyl, 1-naphthyl, and 2-naphthyl) both as to the orientation of the three isopropyl groups and the conformation about the silicon-aromatic bonds. The report comprises dynamic NMR studies of conformational interconversions in solution and in the solid state as well as molecular mechanics calculations. The barriers for the stereomutation processes measured in the crystalline state were found significantly higher than those in solution. PMID- 10814147 TI - Combinatorial Chemistry for Ligand Development in Catalysis: Synthesis and Catalysis Screening of Peptidosulfonamide Tweezers on the Solid Phase. AB - On the basis of a pyrrolidine tweezer 1, a library of peptidosulfonamide tweezers (15a-e, 16a-e) was synthesized on the solid phase. This library was screened in a simultaneous substrate screening procedure for the ability to enantioselectively catalyze the Ti(O-i-Pr)(4)-mediated addition of diethylzinc to aldehydes. One of the best solid-phase tweezer catalyst (i.e., 16d, giving an ee of 32% in solid phase catalysis) was resynthesized in solution (compounds 20 and 21). The now homogeneous solution-phase catalysis showed even better enantioselectivity (i.e., up to 66%). PMID- 10814148 TI - Extremely Efficient Chiral Induction in Conjugate Additions of p-Tolyl alpha Lithio-beta-(trimethylsilyl)ethyl Sulfoxide and Subsequent Electrophilic Trapping Reactions. AB - Reaction of p-tolyl alpha-lithio-beta-(trimethylsilyl)ethyl sulfoxide with alpha,beta-unsaturated esters gave the conjugate addition products as a single diastereomer. The intermediate enolates were subsequently trapped with various alkyl halides or aldehydes to give the products with extremely high stereoselectivity. The reaction with alpha,beta-unsaturated ketones also proceeded with high diastereoselectivity. Protolysis of the enolates derived from the alpha-methyl-alpha,beta-unsaturated esters gave the products with high stereoselectivity. The stereo- and regioselective elimination of the sulfinyl group gave chiral homoallylic carboxylates. PMID- 10814149 TI - Palladium-Catalyzed Synthesis of 1,3-Dienes from Allenes and Organic Halides. AB - A wide range of aryl and vinylic halides react with 1,1-dimethylallene (2a) and potassium carbonate in the presence of Pd(dba)(2) (dba = dibenzylideneacetone) in N,N-dimethylacetamide (DMA) at temperature 100-120 degrees C to give the corresponding dienes CH(2)C(CH(3))CRCH(2) (3a-o), where R is aryl or vinylic, in good to excellent yields. Higher yields of diene products were obtained for aryl bromides than for the corresponding aryl iodides and chlorides. Under similar reaction conditions, tetramethylallene (2b), 1-methyl-1-phenylallene (2c), 1 methyl-3-phenylallene (2d), and 1-cyclohexylallene (2e) also react with aryl and vinylic halides to give diene products (3p-w). For 2d, both E and Z isomers 3t and 3u of the diene product were observed. For 2e, two regioisomers 3vand 3w were isolated with 3w likely from alkene isomerization of 3v. Various palladium systems were tested for the catalytic activity of diene formation. In addition to Pd(dba)(2)/PPh(3), Pd(OAc)(2)/PPh(3), PdCl(2)(PPh(3))(2), and PdCl(2)(dppe) are also very effective as catalysts for the reaction of 2a with p-bromoacetophenone (1a) to give 3a. Studies on the effect of solvents and bases show that DMA and K(2)CO(3) are the solvent and base that give the highest yield of diene 3a. Possible mechanisms for this catalytic diene formation are proposed. PMID- 10814150 TI - Porphyrazinediols: Synthesis, Characterization, and Complexation to Group IVB Metallocenes. AB - The first metalated porphyrazinediols 11 have been prepared from (L)-(+)-dimethyl tartrate via conversion into the corresponding dispoke or 2,3-dimethoxy-2,3 butanediyl protected 2,3-dihydroxymaleonitrile, Linstead macrocyclization, transmetalation, and deprotection. Their stability is very dependent on the nature of the metal ion in the cavity of the porphyrazine. Reaction of these porphyrazinediols with metallocene dichlorides led to new solitaire porphyrazines 12 while DDQ oxidation followed by trapping with diaminomaleonitrile afforded new porphyrazine dinitriles 14. PMID- 10814151 TI - Coupling Reactions of Alkynylsilanes Mediated by a Cu(I) Salt: Novel Syntheses of Conjugate Diynes and Disubstituted Ethynes. AB - Reaction of 1-trimethylsilylalkyne with copper(I) chloride in a polar solvent, DMF, at 60 degrees C under an aerobic conditions smoothly undergoes homo-coupling to give the corresponding symmetrical 1,3-butadiynes in 70-99% yields. In addition, (arylethynyl)trimethylsilanes are found to couple with aryl triflates and chlorides in the presence of Cu(I)/Pd(0) (10 mol %/5 or 10 mol %) cocatalyst system to give the corresponding diarylethynes in 49-99% yields. The cross coupling reaction is applied to a one-pot synthesis of the corresponding unsymmetrical diarylethynes from (trimethylsilyl)ethyne via sequential Sonogashira-Hagihara and the present cross-coupling reactions using two different aryl triflates. The reactions of (arylethynyl)trimethylsilanes with aryl(chloro)ethynes in the presence of 10 mol % of CuCl also yield the corresponding unsymmetrical 1,3-butadiynes in 43-97% yields. PMID- 10814152 TI - Terminal Alkyne-Ethylene Cross-Metathesis: Reaction of 1-Substituted Propargyl Esters at Elevated Ethylene Pressure. AB - Substituted propargylic esters, resistant to complete ethylene cross-metathesis at ambient pressure, underwent cross-metathesis with ethylene at elevated pressure (4 atm) to give 2-substituted butadienes in good to excellent yields. Enantioenriched propargylic acetates, obtained through enzymatic kinetic resolution of secondary propargyl alcohols, similarly underwent ethylene metathesis with retention of stereochemistry at the chiral center. PMID- 10814153 TI - Studies of the Anodic Oxidation of 1,4-Diazabicyclo AB - The title compound (1) was studied at platinum and gold electrodes in acetonitrile. A reversible oxidation peak occurs at +0.30 V vs the standard potential for ferrocenium ion/ferrocene. This process is followed by a second irreversible anodic peak that is due to the oxidation of the initially formed radical cation to the dication. The principal ultimate product of the first oxidation, the conjugate acid of 1, is also oxidized over the range of potentials corresponding to the second anodic peak. The rate of disappearance of the radical cation of 1 has been determined by cyclic voltammetry. The results are best interpreted in terms of parallel pseudo-first-order decay (k(1) = 0.6 s(-)(1)) and second-order reactions. The first of these second-order reactions is either proton transfer from the radical cation to neutral 1 or hydrogen atom abstraction by the radical cation from neutral 1, reactions that give the same products (k(2) = 100 M(-)(1) s(-)(1)) and are kinetically indistinguishable. The other second order reaction is the hydrogen-atom-transfer disproportionation of the radical cation giving the conjugate acid of 1 and the immonium ion (k(3) = 100 M(-)(1) s( )(1)). Both second-order processes must be included to account for the results. The present results are thought to be the first experimental evidence for the occurrence of hydrogen-atom-transfer disproportionation of amine radical cations. PMID- 10814154 TI - Synthesis and Physical Properties of Sterically Congested Cycloalkenes, 1,2-Di tert-butyl-3,3,5,5-tetramethylcyclopentene and 1,2-Di-tert-butyl-3,3,6,6 tetramethylcyclohexene. AB - Two sterically congested cycloalkenes (9 and 10), congeners of tetra-tert butylethylene, were synthesized and characterized. Oxidation of the bicyclic 1,3 dithietane 8 with dimethyldioxirane (DMD) gave the endo,endo-disulfoxide 13, thermal isomerization of which to the endo,exo-disulfoxide 15 followed by oxidation with DMD gave the trioxide 18. Heating 18 in refluxing 1,3-dimethyl-2 imidazolidinone furnished 1,2-di-tert-butyl-3,3,5,5-tetramethylcyclopentene (9) in 69% yield by a 2-fold extrusion process. The reaction of the 1,6-diketone dihydrazone 23 with Se(2)Cl(2) gave the selenadiazoline 34 and the 1,3 diselenetane 35. Heating 34 at 115-130 degrees C gave 1,2-di-tert-butyl-3,3,6,6 tetramethylcyclohexene (10), a "didehydro" derivative of tetra-tert butylethylene, in 43% yield. The C=C bond in 10 is strained in degree comparable to those of most strained alkenes reported so far. PMID- 10814155 TI - Thermolysis and Photosensitized Oxygenation of Tetrasubstituted Cyclopropenes. AB - Bicyclic cyclopropenes 14a, 14b, and 26 were prepared by various synthetic routes. Polymer rose Bengal (p-RB) photosensitized oxygenation of bicyclooctenes 14a,b in CDCl(3) proceeded sluggishly (variable O(2) uptake of ca. 0.35-0.75 equiv in 8 h) and was accompanied by sensitizer bleaching. Preparative gas chromatography of the complex product mixtures from 14a and 14b yielded both dienes (Z- and E-29, 30, and 31) and enones (E- and Z-12, 32, 34). By contrast, p RB photosensitized oxidation of bicyclononene 26 in CDCl(3) proceeded somewhat more rapidly (O(2) uptake of ca. 1 equiv in 2.5 h) yielding enones (20, 42-45) exclusively upon GC separation. The diene products, observed in the case of 14, result from the thermolysis of the remaining unreacted cyclopropenes, while the enones are the oxygenation products. The oxygenation was slowed by radical inhibitors, but not by (1)O(2) quenchers; nor were any oxidation products observed when these cyclopropenes were reacted with triphenylphosphine ozonide, a chemical (1)O(2) source. The data indicates that a photosensitizer-initiated free radical autoxidative process is involved. Likely intermediates in this oxygenation are epoxide 27 or 37 and hydroperoxide 28 or 38, for the bicyclooctene (14) and bicyclononene (26) systems, respectively. The absence of (1)O(2) product in these cyclopropene systems, in contradistinction to their higher homologues, may be attributable to either the relatively long C(alpha) H(allylic) distance in alkylcyclopropenes, which places the abstractable allylic hydrogen "out of reach", or their relatively high IP. Either, or both, of these factors may have slowed the rate of the singlet oxygenation of the cyclopropenes to a point where free radical processes compete favorably. In the course of this study, we also explored the singlet oxygenation (DABCO inhibited) of enones 12a,b and 20. These generated, respectively, a mixture of peroxides identified as alpha keto hydroperoxides 51/54 and hemiperketals 52/55 (the cyclic form of beta-keto hydroperoxides 53/56). Phosphine reduction of these peroxides yields the corresponding alcohols 33/43 and 32/42. PMID- 10814156 TI - Benzocyclobutene: The Impact of Fusing a Strained Ring onto Benzene. AB - The gas-phase acidities of the two aromatic sites in benzocyclobutene were measured in a Fourier transform mass spectrometer using a kinetic technique (i.e., the DePuy method). Fusion of a cyclobutane ring onto benzene is found to have a slight acidifying effect at the alpha-position (3.2 +/- 1.7 kcal mol( )(1)) and little, if any, influence on the beta-site (0.8 +/- 1.9 kcal mol( )(1)). Energetic data (DeltaH degrees (acid) = 386.2 +/- 3.0 kcal mol(-)(1), EA = 0.84 +/- 0.11 eV, and C-H BDE = 92 +/- 4 kcal mol(-)(1)) for the benzylic position were obtained via the bracketing technique and application of a thermodynamic cycle. Differences in the reactivities of the three conjugate bases also were explored. Ab initio and density functional theory calculations were carried out to provide geometries, energies, and insights into the carbanions' electronic structures. PMID- 10814157 TI - Interception of Deaminatively Generated Benzyl Carbenium Ions by Acetone. AB - Essentially free benzyl carbenium ions were generated via protonation of phenyldiazomethane with benzoic acid in acetone. Interestingly, no proton transfer occurred below -20 degrees C. After protonation and dediazoniation of the diazoalkane at -20 degrees C, the solvent was found to intercept the deaminatively generated carbocations to yield initially the corresponding O benzyl oxonium ion and benzyl benzoate. The onium ion, however, was labile under the reaction conditions, and decomposed into a cascade of products whose concentrations as a function of time were used to trace the reaction pathway. Thus, the O-benzyl oxonium ion reacted with benzoate ion to yield (2 benzyloxy)isopropyl benzoate; subsequent decomposition of this O-benzyl-O-benzoyl ketal produced 2,2-dibenzyloxypropane (a dibenzyl ketal), 2-benzyloxypropene, and benzyl alcohol. In a related study, benzyl cations were generated via thermolyses of N-benzyl-N-nitroso-O-benzoyl hydroxylamine at 0 and -70 degrees C. The product distributions were found to be temperature-dependent and different from that in the PhCHN(2) + PhCO(2)H case. PMID- 10814158 TI - A Three-Center Orbital Interaction in the Diels-Alder Reactions Catalyzed by Lewis Acids. AB - Ab initio calculations were performed on title reactions between butadiene and acrolein with BCl(3), AlCl(3), GaCl(3), InCl(3), ZnCl(2), SnCl(2), and SnCl(4). A dimethyl ether molecule is explicitly considered in various reaction systems to examine solvent effects. First, the reaction path of an AlCl(3)-promoting reaction was examined thoroughly. This reaction has two channels. The first one involves a weak reactant-like complex (precursor) and a normal [4 + 2] addition. The second does three elementary processes, one-center addition, ring closing, and Claisen shift. The first channel is more favorable by 12.1 kcal/mol (B3LYP/6 311+G(2d,p) SCRF//B3LYP/6-31G SCRF) than the second one. Then the first channels with other Lewis acids were traced with and without an ether molecule. The ether molecule has an appreciable effect not on geometries but on activation energies. BCl(3) is desolvated and has extraordinarily strong catalytic ability. Even with the strongest catalyst, not a [2 + 4] but a normal [4 + 2] cycloaddition takes place. Except for BCl(3), SnCl(4) is the strongest Lewis acid with the ether molecule. The frontier orbital, LUMO, of acrolein is distorted in the course of the reaction so that the formation of two C-C covalent bonds is possible. The precursor formation and the one-center addition were discussed also by the frontier orbital theory. PMID- 10814159 TI - Why (1S)-Camphanates Are Excellent Resolving Agents for Helicen-1-ols and Why They Can Be Used to Analyze Absolute Configurations. AB - The questions considered in this paper are why, as agents for resolving helicenols, camphanate esters are particularly effective, and why, in all 19 examples studied, when the (1S)-camphanates of (P)- and (M)-helicen-1-ols are chromatographed on silica gel, the former has the lower R(f)(). Models are proposed for the favored conformations of the esters, and to support the models, evidence is provided from five X-ray diffraction analyses and four ROESY analyses supplemented by molecular mechanics calculations. The essential discovery is that, presumably to avoid a steric interaction between a methyl on the camphanate's bridge and the helicene skeleton, the O=CCO conformation is anti periplanar in (M)-helicenol camphanates and syn-periplanar in (P)-helicenol camphanates. In the former, the lactone carbonyl points toward the helicene ring system, and in the latter, it points away. PMID- 10814160 TI - A Novel Outcome of the Hydroperoxide Rearrangement. PMID- 10814161 TI - Synthesis and Reactions of Optically Active Secondary Dialkylphosphine-Boranes. PMID- 10814162 TI - A Divergent, Solid-Phase Approach to Dendritic Ligands on Beads. Heterogeneous Catalysis for Hydroformylation Reactions(1)(a). PMID- 10814163 TI - Masked Formylation with 2-Benzotriazolyl-1,3-dioxolane, a Novel Formyl Cation Equivalent. PMID- 10814165 TI - Spontaneous Addition of Active Methine Compounds to Enol Ethers and alpha,beta Unsaturated Ketones in Aprotic Polar Solvent. PMID- 10814164 TI - pi-Complexes as "Modulators" of Bromine Atom Reactivity in Solution. PMID- 10814166 TI - Conformational Pseudopolymorphism and Solid-State CPMAS NMR Studies for Determination of Solvent-Dependent Solution-State Conformational Preferences for (-)-Scopolamine Hydrobromide/Hydrochloride Salts. PMID- 10814167 TI - Recent Reviews. 56. AB - Reviews are listed in order of appearance in the sources indicated. In multidisciplinary review journals, only those reviews which fall within the scope of this Journal are included. Sources are listed alphabetically in three categories: regularly issued review journals and series volumes, contributed volumes, and other monographs. Titles are numbered serially, and these numbers are used for reference in the index. Major English-language sources of critical reviews are covered. Encyclopedic treatises, annual surveys such as Specialist Periodical Reports, and compilations of symposia proceedings are omitted. This installment of Recent Reviews covers principally the latter part of the 1999 literature. Previous installment: J. Org. Chem. 2000, 65(1), 267-74. PMID- 10814168 TI - Stereocontrolled Synthesis of Key Advanced Intermediates toward Simplified Acetogenin Analogues. PMID- 10814169 TI - Hybrid anthracyclines from a genetically engineered Streptomyces galilaeus mutant. AB - The genetic engineering of antibiotic-producing Streptomyces strains is an approach that is emerging and ready to become established as a successful methodology in developing analogues of the original, pharmaceutically important, natural products obtained from the organisms. The current report highlights this succes by demonstrating the high-level production of novel anthracyclines. The biosynthetic pathways of the nogalamycin-producing Streptomyces nogalater and the aclacinomycin-producing S. galilaeus were combined by transferring the genes of S. nogalater polyketide synthetase into a nonproducing S. galilaeus mutant. The resulting anthracycline antibiotics that were produced possessed structural features characteristic of compounds from both of the undoctored Streptomycesstrains. PMID- 10814170 TI - Additions of enantiopure alpha-sulfinyl carbanions to (S)-N-sulfinimines: asymmetric synthesis of beta-amino sulfoxides and beta-alphamino alcohols AB - The addition of the lithium anions derived from (R)- and (S)-methyl and -ethyl p tolyl sulfoxides to (S)-N-benzylidene-p-toluenesulfinamide provides an easy access route to enantiomerically pure beta-(N-sulfinyl)amino sulfoxides. Stereoselectivity can be achieved when the configurations at the sulfur atoms of the two reagents are opposite (matched pair), thus resulting in only one diastereoisomer, even for the case in which two new chiral centers are created. The N-sulfinyl group primarily controls the configuration of the carbon bonded to the nitrogen, whereas the configuration of the alpha-sulfinyl carbanion seems to be responsible for the level of asymmetric induction, as well as for the configuration of the new stereogenic C-SO carbon in the reactions with ethyl p tolyl sulfoxides. An efficient method for transforming the obtained beta-(N sulfinyl)amino sulfoxides into optically pure beta-amino alcohols, based on the stereoselective non-oxidative Pummerer reaction, is also reported. PMID- 10814171 TI - Further studies of the thermal and photochemical diels-alder reactions of N methyl-1,2,4-triazoline-3,5-dione (MeTAD) with naphthalene and some substituted naphthalenes AB - MeTAD thermally reacted with naphthalene (2) and methylated naphthalenes to give equilibrium mixtures of starting materials and [4 + 2] cycloadducts. Methyl substitution on the naphthalene ring generally increased both the amount of cycloadduct formed and the rate of cycloaddition relative to 2. The isolated cycloadducts were all thermally labile and quantitatively reverted to the parent naphthalene in the presence of 2,3-dimethyl-2-butene as a trap for liberated MeTAD. The rates of the cycloreversion reactions were affected by substitution patterns but not appreciably by solvent. A mechanism for the cycloaddition reaction is presented that proposes the involvement of a charge-transfer complex. Photochemically, MeTAD demonstrated lower regioselectivity in its reactions with substituted naphthalenes relative to the corresponding thermal reactions. PMID- 10814172 TI - Molecular design of crown ethers. 19.(1) synthesis of novel disulfide- and diselenide-bridged Bis(benzo-12-crown-4)s and their Ag(+)-selective electrode properties AB - Two novel heteroatom-bridged bis(benzo-12-crown-4 ether)s, i.e., bis(2-nitro-4,5 (1,4,7,10-tetraoxadecamethylene)phenyl) disulfide (4) and diselenide (5), have been synthesized and characterized by elemental analysis and mass, IR, UV, and (1)H NMR spectroscopy. An X-ray crystallographic structure was obtained for 4. Ion-selective electrodes (ISE) for Ag(+), containing 4 and 5 in PVC membrane as neutral carriers, were prepared, and their selectivity coefficients for Ag(+) () were determined against other heavy metal ions, alkali and alkaline-earth metal ions, and ammonium ion using the matched potential method. These ISEs showed excellent Ag(+) selectivities, log /= -1.2). These values are comparable to those reported for the representative Ag(+)-selective thioethers 6 and 7, revealing that both disulfide and diselenide functionalities in 4 and 5 are equally effective Ag(+)-selective binding sites as the 1,7-dithia-4-oxa functionality in 6 and 7, irrespective of the different atom type and relative position of the sulfur/selenium donors in the ligands. Also discussed are the steric and electronic effects of the nitro groups in 4 and 5 on the Nernstian slopes obtained with the 4- and 5-based ISEs. PMID- 10814174 TI - Synthesis of (-)-7-epiaustraline and (-)-1-epicastanospermine. AB - Highly efficient and selective syntheses of the title compounds are described. The cornerstone of the synthetic plan is the tandem inter [4 + 2]/inter [3 + 2] cycloaddition process. These syntheses differ from previous applications of this strategy in that they incorporate an alkylation in the hydrogenolysis step to close the second ring of the azabicyclic systems. Notable features of the sequence are (1) the highly regio- and stereoselective [3 + 2] cycloaddition of nitronate 15 with siloxymethyl (Z)-beta-silylvinyl ketone (Z)-22b and (2) the highly selective reduction of the resulting ketone 24a with L-Selectride. A single-crystal X-ray structure analysis of synthetic (-)-7-epiaustraline confirmed that the targeted structure was successfully synthesized. This stimulated a reexamination of the structural assignment of the natural product. ( )-1-Epicastanospermine was synthesized in four steps from the common intermediate 27a. The absolute configuration of (-)-1-epicastanospermine was assured by single crystal X-ray structure analysis of intermediate (-)-27a. Thus, the sign of the optical rotation had to be revised. The overall efficiency of these syntheses were 9 steps and 23% yield for (-)-7-epiaustraline and 10 steps and 20% yield for (-)-1-epicastanospermine PMID- 10814173 TI - Synthesis of (+)-casuarine. AB - The first synthesis of (+)-casuarine ((+)-6), a pentahydroxy pyrrolizidine alkaloid of the alexine/australine subclass, is described. The key step is a tandem [4 + 2]/[3 + 2] nitroalkene cycloaddition involving nitrobenzoate 13, chiral vinyl ether 16c, and vinyl silane 10, which establishes five of the six stereocenters present in this potent glycosidase inhibitor. The completion of the synthesis requires only four additional steps to deliver the final product in 20% overall yield. PMID- 10814175 TI - NaY zeolite as host for the selective heterogeneous oxidation of silanes and olefins with hydrogen peroxide catalyzed by methyltrioxorhenium AB - The methyltrioxorhenium(MTO)-catalyzed oxidation of silanes to silanols and the epoxidation of various olefins by aqueous 85% H(2)O(2) proceed in high yields and excellent product selectivities (no disiloxanes, diols) in the presence of the zeolite NaY. The oxidative species is located inside the 12-A supercages. This prevents the bimolecular condensation of the silanol to disiloxane by steric means and the Lewis-acid assisted hydrolysis of the epoxide to the diol. PMID- 10814176 TI - 4,4-Difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) dyes modified for extended conjugation and restricted bond rotations. AB - Five new, constrained, aryl-substituted 4,4-difluoro-4-bora-3a, 4a-diaza-s indacene (BODIPY) dyes (3f,g and 4h-j) were prepared and investigated to see if they have more favorable fluorescence characteristics than the unconstrained systems 2 that were prepared in previous studies. Dye types 3 and 4 have relatively rigid conformations caused by the heteroatom (3f and 3g) or ethylene bridge (4h-j) linkers that preclude free rotation of the substituted-benzene molecular fragments. In the event, the new dye types 3 and 4 have longer lambda(max abs) (620-660 nm) and lambda(max)(fluor) (630-680 nm) values than compounds 2. They also exhibit higher extinction coefficients (>100 000 M(-1) cm( 1), except for 3g). Their fluorescent quantum yields are high (up to 0.72 for 4j), with the exception of compound 3g, which has a quantum yield of only 0.05. The redox properties of dyes 3 and 4 have also been examined. PMID- 10814178 TI - Thermal isomerizations of ketenimines to nitriles: evaluations of sigma-Dot (sigma(*)) constants for spin-delocalizations AB - Rate constants (k(Y)) of the isomerizations of 11 diphenyl N-(substituted benzyl) ketenimines were measured at 40, 50, 60, and 70 degrees C. Activation parameters DeltaH()(Y) and DeltaS()(Y) were obtained using the Eyring equation. The relative rates (k(Y)/k(H)) were fitted into Hammett single correlations (log k(Y)/k(H) = rhosigma and log k(Y)/k(H) = rho(*)sigma(*)). The single correlations have been compared with Hammett dual correlations (log k(Y)/k(H) = rhosigma + rho(*)sigma(*) ). Separate treatments of para and meta substituents yielded even better correlations. Para substituents control the rates through spin delocalizations and inductive effects. The former outweighs the latter when the latter exerts a modest but distinct influence on the rates. On the other hand, inductive effects are the "major" or the sole interactions triggered by meta substituents. PMID- 10814177 TI - Engineering acyclic stereocontrol in the alkylation of vinylglycine-derived dianions: asymmetric synthesis of higher alpha-vinyl amino acids. AB - A generalizable synthesis of higher L-alpha-vinyl amino acids is presented. The strategy pursued here involves the introduction of the amino acid side chain via the alkylation of a chiral, vinylglycine-derived dianionic dienolate, bearing the (-)-8-(beta-naphthyl)menthyl (d'Angelo) auxiliary. A model is presented that postulates a favored "exo-entended" conformation for this dienolate, leading to C(alpha)-alkylation at the si face. The model invokes internal amidate chelation to control ester enolate geometry and soft-soft interactions between the polarizable beta-naphthyl ring of the auxiliary and the extended pi-system of the dienolate to shield the re face. Heats of formation for four conformers of this dianion were calculated for their semiempirical optimized geometries (PM3). The results support the notion that in these vinylglycine-derived dianionic dienolates, "exo" conformations are considerable lower in energy than their "endo" counterparts, with the "exo-entended" conformation being most favorable. In fact, the d'Angelo auxiliary gives a greater degree of acyclic stereocontrol in this system when compared with the (-)-8-phenylmenthyl (Corey) and trans-2 (beta-naphthyl)cyclohexyl auxiliaries, using isobutyl iodide and benzyl bromide as model electrophiles. These dianions are generated from the corresponding dehydrobutyrine esters via sequential deprotonation with LDA and n-BuLi (2 equiv). When alkylations are carried out at -78 degrees C in THF-HMPA, they proceed in 65-81% yields, with both regiocontrol (deconjugative alpha-alkylation is preferred over gamma-alkylation) and a great degree of acyclic stereocontrol [91:9 to >/=98:2 diastereomeric ratios (10 examples)]. The auxiliary may be recovered in high yield (generally 90%) using a modification of Gassman's "anhydrous hydroxide" conditions, in which considerably higher temperatures are employed. Among the side chains introduced directly are those of butyrine, leucine, ornithine, phenylalanine, aspartate, valine, and norvaline. The lysine side chain is elaborated via a 4-step sequence from the alkylation product obtained with 1-chloro-4-iodobutane as electrophile. Importantly, to our knowledge, this work represents the first asymmetric synthesis of L-alpha-vinyl analogues of m-tyrosine, ornithine, and lysine, known time-dependent inhibitors for amino acid decarboxylases. PMID- 10814179 TI - Studies on the synthetic compatibility of aryloxime linkers in the solid-phase synthesis of 3-aminobenzisoxazoles. AB - Further exploration of the scope of our solid-phase method for the synthesis of 3 aminobenzisoxazoles (using the Kaiser oxime resin 1) is described. The effects of base, leaving group, and solvent on the nucleophilic aromatic substitution based resin-loading reaction are discussed. Representative aryloxime intermediates were subjected to a variety of acidic conditions commonly used in protecting group removal to establish the acid stability profile of this linker. Regioselectivity was evaluated with various di- and trifluorobenzonitriles, which gave single benzisoxazole products after loading and cyclorelease reactions. Substituent effects observed in the course of the acid stability and regioselectivity studies suggest that the nitrile plays a critical role in the oxime hydrolysis mechanism. Finally, to establish the compatibility of the aryloxime linker with a variety of useful on-resin synthetic transformations, functionalized substrates were loaded onto resin 1, and carbon-nitrogen, carbon-oxygen, and carbon-carbon bond-forming reactions were successfully executed. PMID- 10814180 TI - Synthesis and characterization of pyridine-based polyamido-polyester optically active macrocycles and enantiomeric recognition for D- and L-amino acid methyl ester hydrochloride. AB - Five new chiral macrocycles, 3a-e, have been prepared by the acylation cyclization of chiral diamine dihydrobromide intermediates 2a-c with 2,6 pyridinedicarbonyl dichloride in highly diluted solution at room temperature. The chiral diesters 1a-c needed for the preparation of the macrocycles were obtained from condensation of corresponding N-(Z)-L-amino acids and 2,6-bishydroxymethyl pyridine in the presence of DCC and DMAP. The enantiomeric recognition of chiral macrocycles 3a-e for D- and L-amino acid methyl ester hydrochlorides has been characterized by fluorescence spectra, which indicate that some of them exhibited significant chiral recognition for the enantiomers of D- and L-amino acid methyl ester hydrochlorides. The stoichiometry and binding constants of 3a-L-Am(2) and 3c-L-Am(2) complexes have been determined. An X-ray analysis of the chiral macrocycle 3b show that the chiral ligand is rather rigid and strained. PMID- 10814181 TI - Glucose-derived 3'-(carboxymethyl)-3'-deoxyribonucleosides and 2', 3'-lactones as synthetic precursors for amide-linked oligonucleotide analogues. AB - Treatment of a 1,2-O-isopropylidene-3-ketopentofuranose derivative (obtained from D-glucose) with [(ethoxycarbonyl)methylene]triphenylphosphorane and catalytic hydrogenation of the resulting alkene gave stereodefined access to 3 (carboxymethyl)-3-deoxy-D-ribofuranose derivatives. Esters of 5-O-acetyl- or 5 azido-5-deoxy-3-(carboxymethyl)-D-ribofuranose were coupled with nucleobases to give branched-chain nucleoside derivatives. Ester saponification and protecting group manipulation provided 2'-O-(tert-butyldimethylsilyl) ethers of 5'-azido-5' deoxy- or 5'-O-(dimethoxytrityl) derivatives of 3'-(carboxymethyl)-3' deoxyribonucleosides that are effective precursors for synthesis of amide-linked oligoribonucleosides. PMID- 10814182 TI - Quantitative monitoring of solid phase organic reactions by high-resolution magic angle spinning NMR spectroscopy AB - Three possible high-resolution magic angle spinning (HR MAS) NMR experiments to quantitatively monitor a solid phase supported Horner-Emmons reaction are presented. In the first experiment we follow the solid phase reaction in deuterated solvent directly in the NMR rotor. The second quantification is done by reconditioning of a few milligrams of resin from an undefined reaction vessel by washing, drying, and reswelling in deuterated solvent, and the evaluation of the amount of resin bound structures by comparing to an external standard. The third experiment represents the first analytical quantification of resin-bound structures without any sample preparation, except the transfer of resin-solvent suspension (large excess of reagents in protonated dimethylformamide) from the reaction vessel to the NMR rotor. PMID- 10814183 TI - Total synthesis of cyclosporin O both in solution and in the solid phase using novel thiazolium-, immonium-, and pyridinium-type coupling reagents: BEMT, BDMP, and BEP. AB - Cyclosporin O (1), an extensively N-methylated immunosuppressive cyclic undecapeptide isolated from Tolypocladium inflatum Gams, was synthesized in 20 23% overall yield via 4 + 7 segment condensation and cyclization by the combined utilization of novel thiazolium- and immonium-type peptide coupling reagents 2 bromo-3-ethyl-4-methyl thiazolium tetrafluoroborate (BEMT) and 5-(1H-benzotriazol 1-yloxy)-3,4-dihydro-1-methyl 2H-pyrrolium hexachloroantimonate (BDMP) as well as compound 2-bromo-1-ethyl pyridinium tetrafluoroborate (BEP). BEMT and BEP, which have been proven to be very efficient for the coupling of peptide segments containing N-alkylated amino acid residues with respect to the fast reaction speed, low racemization, and high yields, were used to construct hindered amide bonds in CsO with the addition of HOAt, whereas the most efficient HOBt-derived immonium type reagent, BDMP, was used to perform the coupling of coded amino acids in CsO. Thus, the highly hindered protected 8-11 tetrapeptide 25 was successfully synthesized using BEMT in 65% yield, and the 1-7 heptapeptide 21 was obtained in 52-55% yield by the rationally combined utilization of BDMP, BEMT, and BEP. The synthesis of the linear undecapeptide 27 of CsO in the solid phase using BEMT and BEP was accomplished for the further evaluation of the effectiveness of these reagents. PMID- 10814184 TI - Chemical synthesis of cross-link lesions found in nitrous acid treated DNA: a general method for the preparation of N2-substituted 2'-deoxyguanosines. AB - Treatment of DNA with nitrous acid results in the formation of DNA-DNA cross links. Two cross-link lesions have previously been isolated and their structures assigned based on spectroscopic data. The major lesion has been proposed to consist of two deoxyguanosine (dG) nucleosides sharing a common N2 atom (1), while the structure of the minor lesion has been proposed to consist of a common nitrogen atom linking C2 of a dG nucleoside to C6 of deoxyadenosine (2). The chemical synthesis of 1 and 2, utilizing a palladium-catalyzed coupling, is described herein. It is demonstrated that the spectroscopic properties of synthetic 1 are identical to that of lesion 1 obtained from nitrous acid cross linked DNA, thus providing a proof of its structure. Comparison of the limited spectroscopic data available for lesion 2 originating from nitrous acid cross linked DNA to synthetic 2 supports its structural assignment. The synthetic approach used for synthesis of 1 and 2 is shown to be a general method for the preparation of a variety of N2-substituted dG nucleosides in good yields. PMID- 10814185 TI - The "Non-Oxidative" pummerer reaction: conclusive evidence for S(N)2-type stereoselectivity, mechanistic insight, and synthesis of enantiopure L-alpha trifluoromethylthreoninate and D-alpha-trifluoromethyl-allo-threoninate(1) AB - Enantiopure methyl D-alpha-trifluoromethyl-allo-threoninate 18 and L-alpha trifluoromethylthreoninate 19 were synthesized using (R)-ethyl p-tolylsulfoxide as chiral alpha-hydroxyethyl anion equivalent. The key step was the S(N)2-type replacement of the sulfinyl auxiliary with a hydroxy group, via trifluoroacetic anhydride promoted "non-oxidative" Pummerer reaction (NOPR) of the diastereomeric intermediate beta-sulfinyl amines 14 and 15, obtained by condensation of (R) ethyl p-tolylsulfoxide 13 with the N-Cbz imine of methyl trifluoropyruvate 12. The conclusive evidence for S(N)2-type stereoselectivity of the NOPR was achieved by X-ray diffraction of both the starting diastereomer 14 and the p-bromobenzoate 25, obtained from the threoninate 19. NMR monitoring of the NOPR performed on 15 allowed the detection of a transient intermediate, which was identified as the four membered cyclic sigma-sulfurane 27. This intermediate spontaneously rearranged (40 min, rt) into the corresponding sulfenamide 17, probably via an intramolecular displacement of the sulfinyl by a trifluoroacetoxy group, with inversion of configuration at the carbon stereocenter. The same process occurred for the diastereomeric beta-sulfinyl amine 14, but the sulfenamide 16 was formed at a very fast rate, thus precluding NMR detection of the corresponding sigma sulfurane intermediate 26. One-pot treatment of the diastereomeric sulfenamides 16 and 17 with NaBH(4) afforded very good yields of the corresponding threoninates 18 and 19. PMID- 10814186 TI - NO(2)(+) nitration mechanism of aromatic compounds: electrophilic vs charge transfer process AB - The nitration of methylnaphthalenes with NO(2)BF(4) and NOBF(4) was examined in order to shed light on the controversial aromatic nitration mechanism, electrophilic vs charge-transfer process. The NO(2)(+) nitration of 1,8 dimethylnaphthalene showed a drastic regioselectivity change depending on the reaction temperature, where ortho-regioselectivity at -78 degrees C and para regioselectivity at 0 degrees C were considered to reflect the electrophilic and the direct or alternative charge-transfer process, respectively, because the NO(+) nitration through the same reaction intermediates as in the NO(2)(+) nitration via a charge-transfer process resulted in para-regioselectivity regardless of the reaction temperature. The NO(2)(+) nitration of redox potential methylnaphthalenes higher than 1,8-dimethylnaphthalene gave a similar ortho regioselectivity enhancement to 1,8-dimethylnaphthalene at lower temperature, thus reflecting the electrophilic process. On the other hand, the NO(2)(+) nitration of redox potential methylnaphthalenes lower than 1,8 dimethylnaphthalene showed para-regioselectivity similar to the NO(+) nitration, indicating the direct or alternative charge-transfer process. In the presence of strong acids where the direct charge-transfer process will be suppressed by protonation, the ortho-regioselectivity enhancement was observed in the NO(2)(+) nitration of 1,8-dimethylnaphthalene, suggesting that the direct charge-transfer process could be the main process to show para-regioselectivity. These experimental results imply that the NO(2)(+) nitration proceeds via not only electrophilic but also direct charge-transfer processes, which has been considered to be unlikely because of the high energy demanding process of a bond coordination change between NO(2)(+) and NO(2). Theoretical studies at the MP2/6 31G(d) level predicted ortho- and para-regioselectivity for the NO(2)(+) nitration via electrophilic and charge-transfer processes, respectively, and the preference of the direct charge-transfer process over the alternative one, which support the experimental conclusion PMID- 10814187 TI - Catalytic enantioselective protonation of lithium enolates with chiral imides AB - The catalytic enantioselective protonation of simple enolates was achieved using a catalytic amount of chiral imides and stoichiometric amount of achiral proton sources. Among the achiral proton sources examined in the protonation of the lithium enolate of 2,2,6-trimethylcyclohexanone catalyzed by (S,S)-imide 1, 2, 6 di-tert-butyl-p-cresol (BHT) and its derivatives gave the highest enantiomeric excess. For example, 90% ee of (R)-enriched ketone was obtained when (S,S)-imide 1 (0.1 equiv) and BHT (1 equiv) were used. Use of 0.01 equiv of the chiral catalyst still caused a high level of asymmetric induction. For catalytic protonation of the lithium enolate of 2-methylcyclohexanone, chiral imide 6 possessing a chiral amide portion was superior to (S,S)-imide 1 as a chiral proton source and the enolate was effectively protonated with up to 82% ee. PMID- 10814188 TI - Direct transformations of ketones to gamma-unsaturated thiols via AB - The reaction of a series of ketones with dimsylsodium in dimethyl sulfoxide resulting in the formation of gamma-unsaturated thiols was studied experimentally. [2,3]-Sigmatropic rearrangements of beta-unsaturated sulfinyl carbanions are involved at the key step of those transformations. DFT computations at the B3LYP/6-31+G level indicated that such rearrangements, as well as some typical [2, 3]-sigmatropic rearrangements, e.g., thermal rearrangements of allyl sulfoxides and Wittig rearrangements of sulfur ylides and lithiated allyloxy methyl anions, are concerted and moderately synchronous processes. Negative (diatropic) nucleus-independent chemical shifts (NICS) and high diamagnetic susceptibility exaltations indicate that the transition structures of these [2,3]-sigmatropic migrations are aromatic. PMID- 10814189 TI - Olefin epoxidation by peroxo complexes of cr, mo, and W. A comparative density functional study AB - The epoxidation of olefins by peroxo complexes of Cr(VI), Mo(VI) and W(VI) was investigated using the B3LYP hybrid density functional method. For the mono- and bisperoxo model complexes with the structures (NH(3))(L)M(O)(2)(-)(n)()(eta(2) O(2))(1+)(n)() (n = 0, 1; L = none, NH(3); M = Cr, Mo, W) and ethylene as model olefin, two reaction mechanism were considered, direct oxygen transfer and a two step insertion into the metal-peroxo bond. The calculations reveal that direct attack of the nucleophilic olefin on an electrophilic peroxo oxygen center via a transition state of spiro structure is preferred as significantly higher activation barriers were calculated for the insertion mechanism than for the direct mechanism. W complexes are the most active in the series investigated with the calculated activation barriers of direct oxygen transfer to ethylene decreasing in the order Cr > Mo > W. Barriers of bisperoxo species are lower than those of the corresponding monoperoxo species. Coordination of a second NH(3) base ligand to the mono-coordinated species, (NH(3))M(O)(2)(eta(2)-O(2)) and (NH(3))MO(eta(2)-O(2))(2), results in a significant increase of the activation barrier which deactivates the complex. Finally, based on a molecular orbital analysis, we discuss factors that govern the activity of the metal peroxo group M(eta(2)-O(2)), in particular the role of metal center. PMID- 10814190 TI - Halogenations of anthracenes and Dibenz AB - Halogenation of dibenz[a,c]anthracene (1) by NBS in CCl(4) affords the products of 9- and 10-monobromination in the ratio of 9:1. The reaction is accelerated by iodine, and HBr effects rearrangement of 9-bromo product to the sterically less crowded 10-bromo isomer. The mechanism is proposed to involve reversible addition of Br(2), followed by elimination of HBr. Reaction of NCS with 1 in CCl(4) requires addition of HCl and affords exclusively 9-chlorination. The different reactivities of NBS and NCS are ascribed to the relative amounts of free halogen produced (due to differences in N-X bond strengths involving Br and Cl), and the different sizes of the halogens. Under similar conditions, NCS chlorinates 9 bromoanthracene (2a) to afford 9,10-dichloroanthracene and 9-bromo-10 chloroanthracene in the ratio of 65:35. This reaction ostensibly occurs by addition of Cl(2) to 2a, followed by preferential loss of HBr rather than HCl. 9 Methylanthracene (3) affords exclusively 9-(bromomethyl)anthracene with NBS in the absence of iodine, but mainly (67%) 9-bromo-10-methylanthracene in the presence of iodine. Chlorination of 3 with NCS in the presence of HCl also affords mostly (65%) nuclear halogenation. Nuclear bromination of anthracene, 9 methylanthracene, and dibenz[a, c]anthracene by NBS in the absence of added HBr is accelerated by iodine. This effect is probably due to an increase in the amount of bromine produced from NBS in the presence of iodine. PMID- 10814191 TI - Aminosulf(ox)ides as ligands for Iridium(I)-catalyzed asymmetric transfer hydrogenation AB - A new class of efficient catalysts was developed for the asymmetric transfer hydrogenation of unsymmetrical ketones. A series of chiral N,S-chelates (6-22) was synthesized to serve as ligands in the iridium(I)-catalyzed reduction of ketones. Both formic acid and 2-propanol proved to be suitable as hydrogen donors. Sulfoxidation of an (R)-cysteine-based aminosulfide provided a diastereomeric ligand family containing a chiral sulfur atom. The two chiral centers of these ligands showed a clear effect of chiral cooperativity. In addition, aminosulfides containing two asymmetric carbon atoms in the backbone were synthesized. Both the sulfoxide-containing beta-amino alcohols and the aminosulfides derived from 1,2-disubstituted amino alcohols gave rise to high reaction rates and moderate to excellent enantioselectivities in the reduction of various ketones. The enantioselective outcome of the reaction was favorably affected by selecting the most appropriate hydrogen donor. Enantioselectivities of up to 97% were reached in the reduction of aryl-alkyl ketones. PMID- 10814192 TI - Asymmetric and nonasymmetric addition of RLi and RMgX to 3-methoxynaphthalen-2-yl oxazolines and imines. An approach to substituted 2-tetralones AB - Chiral and achiral 3-methoxynaphthalen-2-yl oxazolines 4a,b failed to undergo an aromatic nucleophilic displacement of the 3-methoxy group with organolithium reagents and instead afforded dihydronaphthalenes 9 and 14 in 30-95% yield. Dihydronaphthalenes 9 (racemic) and 14 (nonracemic) were easily converted into the corresponding aldehydes 15. Alternatively, aldehydes 15 were prepared via tandem addition of Grignard reagents to imines 17 in 50-65% yield. Aldehydes 15 served as precursors to 3,3, 4-trisubstituted 2-tetralones 16. Use of methyl chloroformate to trap the azaenolate derived from 17f and i-PrMgCl afforded, in 65% yield, a versatile synthetic intermediate 23 which may serve to access 4 alkyl-, 3,4-dialkyl-, 3,4-disubstituted and 3,3, 4-trisubstituted 2-tetralones with diverse substitution patterns. PMID- 10814194 TI - New chiral didehydroamino acid derivatives from a cyclic glycine template with 3,6-dihydro-2H-1,4-oxazin-2-one structure: applications to the asymmetric synthesis of nonproteinogenic alpha-amino acids. AB - New chiral (Z)-alpha,beta-didehydroamino acid (DDAA) derivatives with 3,5-dihydro 2H-1,4-oxazin-2-one structure 11a-f have been stereoselectively prepared after condensation of chiral glycine equivalent 7 with aldehydes in the presence of K(2)CO(3) under mild solid-liquid phase-transfer catalysis reaction conditions. These new systems have been used in diastereoselective cyclopropanation reactions using Corey's ylide for the asymmetric synthesis of 1-aminocyclopropane-1 carboxylic acids (ACCs) such as allo-corononamic and allo-norcoronamic acids. The hydrogenation reaction of these systems at ambient pressure in the presence of formaldehyde affords saturated oxazinones and N-methylated oxazinones which have been transformed into the N-methyl-alpha-amino acids (N-MAAs) (S)-2 (methylamino)butanoic acid and (S)-N-methylleucine. In addition, the parent alpha, beta-didehydroalanine derivative 11g has been prepared by a direct aminomethylation-elimination sequence from 7 and Eschenmoser's salt and has been used in Diels-Alder cycloaddition with endo selectivity for the synthesis of the enantiomerically pure bicyclic alpha-amino acids (-)-2-aminobicyclo[2.2.1]heptane 2-carboxylic and (-)-2-aminobicyclo[2.2.2]octane-2-carboxylic acids. PMID- 10814193 TI - Design and synthesis of a transferable farnesyl pyrophosphate analogue to Ras by protein farnesyltransferase. AB - The posttranslational addition of a farnesyl moiety to the Ras oncoprotein is essential for its membrane localization and is required for both its biological activity and ability to induce malignant transformation. We describe the design and synthesis of a farnesyl pyrophosphate (FPP) analogue, 8-anilinogeranyl pyrophosphate 3 (AGPP), in which the omega-terminal isoprene unit of the farnesyl group has been replaced with an aniline functionality. The key steps in the synthesis are the reductive amination of the alpha,beta-unsaturated aldehyde 5 to form the lipid analogue 6, and the subsequent conversion of the allylic alcohol 7 to the chloride 8 via Ph(3)PCl(2) followed by displacement with [(n Bu)(4)N](3)HP(2)O(7) to give AGPP (3). AGPP is a substrate for protein farnesyltransferase (FTase) and is transferred to Ras by FTase with the same kinetics as the natural substrate, FPP. AGPP is highly selective, showing little inhibitory activity against either geranylgeranyl-protein transferase type I (GGTase I) (K(i) = 0.06 microM, IC(50) = 20 microM) or squalene synthase (IC(50) = 1000 microM). AGPP is the first efficiently transferable analogue of FPP to be modified at the omega-terminus that provides a platform from which additional analogues can be made to probe the biological function of protein farnesylation. AGPP is the first example of a class of compounds that are alternate substrates for protein isoprenylation that are not inhibitors of squalene synthase. PMID- 10814195 TI - Synthesis of functionalized chiral carbocyclic cleft molecules complementary to Troger's base derivatives. AB - The synthesis of optically pure functionalized cleft molecules derived from dibenzobicyclo[b,f][3.3.1]nona-5a,6a-diene-6,12-dione is reported. These clefts are reminiscent of Troger's base but contain clefts with different dimensions and additional carbonyl (or alcohol) groups that may be utilized in molecular recognition studies. The 2,8-dimethyl and 2,8-dibromo derivatives were synthesized via an intramolecular Friedel-Crafts acylation and were resolved by chiral HPLC. The 2,8-dinitro derivative was prepared by regiospecific nitration of dibenzobicyclo[b,f][3.3.1]nona-5a, 6a-diene-6,12-dione. The dibromo and dinitro derivatives allow direct access to a range of functionalized molecular clefts. Palladium-catalyzed coupling of the dibromo derivative afforded the disubstituted phenyl, anisole, and acetylene derivatives, while reduction of the dinitro derivative and acetylation provided amino-dione and amide-hydroxyl derivatives. X-ray crystal structures of the dimethyl 12, dibromo 13, di(p methoxyphenyl) 16, dinitro 18, and dimethyl dinitro 22 derivatives show cleft angles between the planes between the aromatic rings of 84-104 degrees. The synthetic route, structural features, and potential for molecular recognition studies of this class of clefts are compared with those of the more widely studied Troger's base cleft molecules. PMID- 10814196 TI - Effect of chain length and ring size of alkyl and cycloalkyl side-chain substituents upon the biological activity of brassinosteroids. Preparation of novel analogues with activity exceeding that of brassinolide. AB - A series of brassinosteroids with different alkyl or cycloalkyl substituents in place of the isopropyl group at C-24 of brassinolide (1) were prepared by the CuCN-catalyzed addition of Grignard reagents to (threo-2R,3S,5alpha,22R,23R,24S) 23,24-epoxy-6, 6-(ethylenedioxy)-2,3-(isopropylidenedioxy)-26, 27-dinorcholestan 22-ol (9), followed by deketalization and Baeyer-Villiger oxidation. Compound 9 was employed as part of a 70:30 threo/erythro mixture of epoxides 9 and 10, from which the erythro-epoxide 10 was recovered intact after the Grignard additions. Thus, the corresponding n-dodecyl, n-hexyl, n-propyl, tert-butyl, cyclohexyl, cyclopentyl, cyclobutyl, and cyclopropyl analogues of brassinolide were obtained. A rearrangement byproduct was observed during the preparation of the cyclopropyl substituted brassinosteroid when ether was used as the solvent in the Grignard reaction, but could be avoided by the use of THF. A method for recycling the undesired erythro-epoxide 10 was developed on the basis of deoxygenation with tellurium and lithium triethylborohydride. The rice leaf lamina inclination assay was then used to measure the bioactivity of the products. In general, increasing activity was observed as the length or ring size of the C-24 hydrocarbon substituent decreased. The novel cyclobutyl- and cyclopropyl-substituted analogues of brassinolide (1) were ca. 5-7 times as active as 1 and thus appear to be the most potent brassinosteroids reported to date. Further enhancement of the bioactivity of all of the above brassinosteroids, except that of the inactive n-dodecyl derivative, was observed when the brassinosteroid was applied together with an auxin, indole-3-acetic acid (IAA). The synergy between the brassinosteroids and IAA thus increased the bioactivity of the brassinosteroids, including the cyclopropyl and cyclobutyl derivatives, by ca. 1-2 orders of magnitude. PMID- 10814197 TI - Electron acceptors of the fluorene series. 10.(1) novel acceptors containing butylsulfanyl, butylsulfinyl, and butylsulfonyl substituents: synthesis, cyclic voltammetry, charge-transfer complexation with anthracene in solution, and X-ray crystal structures of two tetrathiafulvalene complexes AB - 2,4,5,7-Tetranitro-9-fluorenone (1b) reacts readily with n-butanethiol in dipolar aprotic solvents with selective substitution of nitro groups by butylsulfanyl groups in positions 2 and 7 (2, 3); the 2,5-isomer 4 was formed only as a minor product (<1%). Condensation of fluorenones 2-4 with malononitrile yielded 9 dicyanomethylene derivatives 5-7, which showed strong intramolecular charge transfer (lambda approximately 510-560 nm) and were found to sensitize the photoconductivity of carbazole-containing polymer films. Oxidation of sulfides 2 4 gave sulfoxide 8 or sulfones 9-11, which then were converted into their corresponding dicyanomethylene derivatives 12-15. All these novel acceptors showed three reversible single-electron reduction waves (cyclic voltammetry) yielding radical anion, dianion, and radical trianion; moreover, acceptors 13-15 showed also a fourth reduction wave, representing reversible tetraanion formation. Substitution of the oxygen of the carbonyl group in the fluorenones by a dicyanomethylene group increased the thermodynamic stability (K(SEM) growth) of the radical anion; K(SEM) ranged from 3 x 10(5) to 3 x 10(9) M(-1). CV measurements characterize compounds 3, 4 (EA = 1. 86-1.89 eV) as poor acceptors, 2, 6-11 (EA = 2.13-2.31 eV) as moderate acceptors, and 5, 12-15 (EA = 2.53-2.66 eV) as strong electron acceptors. Charge-transfer complex (CTC) formation between acceptors 9, 10, 13, 14, and anthracene as a donor was monitored by the appearance of additional low-energy bands in the visible region (CTC bands) of their electron absorption spectra. Increasing the EA of the acceptors from 9 fluorenones to the corresponding 9-dicyanomethylenefluorenes increases the complexation constants K(CTC) by 2.5-3 times, while sulfonyl substituents present substantial steric hindrance for complexation (as compared to the nitro group), decreasing K(CTC) values. Two CTCs for acceptors 14 and 17 with tetrathiafulvalene (TTF) were obtained, and their structures were solved by single-crystal X-ray diffractometry, giving the stoichometries 14:TTF, 2:3, and 17:TTF:PhCl, 1:1:0.5. In the former complex the packing motif is a mixed.DDAD'A. stack; in the latter complex the D and A moieties form unusually close CT pairs, which pack in a herringbone motif. PMID- 10814198 TI - Efficient synthesis of 3, 6-dideoxy-beta-D-arabino-hexopyranosyl-terminated LacdiNac glycan chains of the Trichinella spiralis parasite. AB - The synthesis of a linear trisaccharide epitope of the Trichinella spiralis N linked glycan, in a form amenable to glycoconjugate formation, is reported. The trisaccharide contains the synthetically challenging LacdiNAc [beta-GalpNAc(1- >4)-beta-GlcpNAc] element, as well as a terminal 3,6-dideoxy-beta-D-arabino hexopyranose (tyvelose) residue. An orthogonal protection strategy is described, which permits the protection and manipulation of three amino groups present in the disaccharide beta-GalNAc(1-->4)-beta-GlcNAc and the tether used to prepare neoglycoconjugates. The beta-linked dideoxyhexose was generated in excellent yield by the introduction of the dideoxyhexose unit as a beta-D-ribo hexopyranoside (paratose) followed by an oxidation-reduction sequence to generate the beta-D-arabino configuration in high diastereomeric excess. The required dideoxyhexose donor was synthesized in a series of high-yielding steps from glucose utilizing the p-methoxyphenyl glycoside. PMID- 10814199 TI - Synthesis of enantiopure trans-3,4-disubstituted piperidines. An enantiodivergent synthesis of (+)- and (-)-paroxetine. AB - Reaction of (R)-phenylglycinol with methyl 5-oxopentanoate gave either bicyclic lactam cis-1 (the kinetic product) or its isomer trans-1 (under equilibrating conditions) as the major products, which were converted to the corresponding (cis or trans) unsaturated lactams 4 and 5. On treatment with lithium alkyl (or aryl) cyanocuprates, these chiral building blocks undergo conjugate addition to give enantiopure trans-3,4-substituted 2-piperidone derivatives in high yield and stereoselectivity. The synthetic potential of this transformation is illustrated by the synthesis of (+)-femoxetine and the two enantiomers of the known antidepressant paroxetine. PMID- 10814200 TI - Synthesis and reactions of (E)- and (Z)-1,3-dibromo-2-methoxypropene AB - (Z)-1,3-Dibromo-2-methoxypropene is prepared in 90% yield by dehydrohalogenation of 1,2,3-tribromo-2-methoxypropane with diisopropylamine in dichloromethane. The E-isomer can be obtained as the only product in almost quantitative yield by UV irradiation of the Z-isomer. Nucleophilic displacement reactions of the allylic bromide and palladium-catalyzed coupling reactions of the vinylic bromide in (E)- and (Z)-1,3-dibromo-2-methoxypropene have been studied. PMID- 10814201 TI - Studies on the mechanism of B(C(6)F(5))(3)-catalyzed hydrosilation of carbonyl functions AB - The strong organoborane Lewis acid B(C(6)F(5))(3) catalyzes the hydrosilation (using R(3)SiH) of aromatic and aliphatic carbonyl functions at convenient rates with loadings of 1-4%. For aldehydes and ketones, the product silyl ethers are isolated in 75-96% yield; for esters, the aldehydes produced upon workup of the silyl acetal products can be obtained in 45-70% yield. Extensive mechanistic studies point to an unusual silane activation mechanism rather than one involving borane activation of the carbonyl function. Quantitative kinetic studies show that the least basic substrates are hydrosilated at the fastest rates; furthermore, increased concentrations of substrate have an inhibitory effect on the observed reaction rate. Paradoxically, the most basic substrates are reduced selectively, albeit at a slower rate, in competition experiments. The borane thus must dissociate from the carbonyl to activate the silane via hydride abstraction; the incipient silylium species then coordinates the most basic function, which is selectively reduced by [HB(C(6)F(5))(3)](-). In addition to the kinetic data, this mechanistic proposal is supported by a kinetic isotope effect of 1.4(5) for the hydrosilation of acetophenone, the observation that B(C(6)F(5))(3) catalyzes H/D and H/H scrambling in silanes in the absence of substrate, computational investigations, the synthesis of models for proposed intermediates, and other isotope labeling and crossover experiments. PMID- 10814202 TI - Cyclization into hydrindanones using samarium diiodide AB - Samarium(II) iodide has been employed to promote vinylogous pinacol coupling reaction of aldehyde onto alpha,beta-unsaturated ketones. The diastereoselectivity of 6-endo products was changed by addition of a proton source and/or HMPA and by the reaction temperature. The cyclization reactions described herein provide a general approach to the syntheses of 3,3 dimethylhydrindanes with a cis-relationship between the OH at C-4 and the proton at C-3a with good diastereoselectivity and under mild reaction conditions. PMID- 10814203 TI - Tandem one-Pot palladium-catalyzed reductive and oxidative coupling of benzene and chlorobenzene AB - The in situ combination of oxidative coupling of benzene to biphenyl and reductive coupling of chlorobenzene (also to biphenyl) using palladium catalysts (Pd(2+)/Pd(0)) is described. In each cycle, the reductive process regenerates the catalyst for the oxidative process and vice versa. Kinetic investigations show that the reaction rate depends on [C(6)H(6)], [C(6)H(6)Cl], and catalyst loading, with E(a)() = 13 kcal mol(-)(1). The reduced palladium catalyst undergoes deactivation through aggregation and precipitation, but it is observed that during this deactivation process the Pd(0) becomes an active catalyst for the reductive coupling of chlorobenzene. Accordingly, while Pd(0)/C particles are inactive, Pd(0) colloids do catalyze the tandem reaction. Conversion is increased in the presence of a phase-transfer catalyst, presumably due to stabilization of the active Pd(0) clusters. The two halves of the catalytic cycle are examined in the light of previous research, regarding analogous oxidative and reductive coupling reactions, using stoichiometric amounts of PdCl(2) and Pd(0), respectively. The roles of homogeneous PdCl(2) and Pd(0) clusters are discussed. PMID- 10814204 TI - Synthesis and reactions of a stable o-quinoid 10-pi-electron system, Furo AB - Methyl 4,6-dichloro-3-(diethylamino)furo[3, 4-c]pyridine-1-carboxylate (6), an intermediate in the Hamaguchi-Ibata reaction involving the Rh(II)-catalyzed intramolecular reaction of a diazo group with the carbonyl of an adjacent amido group, has been isolated and characterized. PM3 calculations reveal the heat of formation (DeltaH(f)) of this remarkably stable molecule to be -77.7 kcal/mol. Compound 6 undergoes a facile Diels-Alder cycloaddition with a variety of dienophiles to give polysubstituted isoquinoline derivatives via ring opening of initially formed cycloadducts. In each case the cycloaddition proceeds with high regioselectivity, with the electron-withdrawing group located ortho to the amino group. The most favorable FMO interaction is between the HOMO of the azaisobenzofuran 6 and the LUMO of the dienophile. The atomic coefficient at the ester carbon of the azaisobenzofuran 6 is larger than the amino center, and this nicely accommodates the observed regioselectivity. PMID- 10814205 TI - 2-Azanorbornyl alcohols: very efficient ligands for ruthenium-catalyzed asymmetric transfer hydrogenation of aromatic ketones AB - 2-Azanorbornyl-derived amino alcohols were prepared and evaluated as ligands in the Ru(II)-catalyzed asymmetric transfer hydrogenation of aromatic ketones. To improve selectivity and rate, the structure of the ligand was optimized. Acetophenone was reduced using 0.5 mol % catalyst in 40 min in 94% ee. This system was also able to reduce a wide range of aromatic ketones to the corresponding alcohols, while maintaining high enantioselectivities and yields. The effects of catalyst loading and the presence of cosolvents in the reaction vessel were examined, and a linearity study was also done. PMID- 10814206 TI - Base-promoted in situ generation of methyl acrylate from dimethyl 3, 3' dithiodipropionate. Application To N-alkylation of heterocycles AB - In basic medium, dimethyl 3,3'-dithiopropionate generates methyl acrylate, which serves in situ as a source of propionate moiety. This property was applied to the alkylation of indoles and other nitrogen heterocycles leading to a series of 3 (heteroaryl-substituted) propionates. A mechanistic rationale for the generation of acrylate is presented along with supportive experimental data. PMID- 10814207 TI - Synthesis of soluble bis-terpyridine ligands bearing ethynylene-phenylene spacers AB - Soluble and rigid terpyridine-based ditopic ligands bearing one to five phenylene/ethynylene modules have been synthesized by way of a stepwise procedure. Each module is attached to the terpyridine unit via an ethynylene fragment and functionalized at the 4-position with an additional ethynylene connector and in the 2,5-positions with two flexible dodecyloxy chains. The synthetic protocol is based on sequential Pd(0)-catalyzed cross-coupling reactions between a terpyridine subunit grafted with the necessary diethynyl/phenyl or ethynylphenyl/bromide appendage. For ditopic ligands displaying an even number of phenyl/ethynylene modules, the final step involves a single cross-coupling reaction between 4'-ethynylene-2,2':6',6' '-terpyridine and the appropriate bromo derivative. In the case of the ligands having an odd number of phenylene/ethynylene fragments, a double cross-coupling reaction between an extended dibromopolyphenylene intermediate and 4'-ethynylene-2,2':6',6' ' terpyridine or 1-(4'-ethynylene-2,2':6',2' '-terpyridine)-4-ethynylene-2,5 didodecyloxy-benzene is required. For ligands I-V, optimal preparative conditions were found with [Pd(0)(PPh(3))(4)] (6 mol %) in n-propylamine at 70 degrees C. Oxidative dimerization of the 1-(4'-ethynylene-2,2':6',2' '-terpyridine)-4 ethynylene-2,5-didodecyloxybenzene derivative in the presence of cupric salts and oxygen gives the corresponding homoditopic ligand II(2)() bearing a central diphenyldiacetylene spacer. Spectroscopic data for the new oligomers are discussed in terms of the extent of pi-electron conjugation. Upon increasing the number of phenylene/ethynylene modules, there is a progressive lowering in energy of absorption and fluorescence transitions. PMID- 10814208 TI - Evaluation of beta- and gamma-effects of group 14 elements using intramolecular competition AB - To evaluate beta-effects and gamma-effects of group 14 elements, we have devised a system in which the intramolecular competition between gamma-elimination of tin and beta-elimination of silicon, germanium, and tin can be examined. Thus, the reactions of alpha-acetoxy(arylmethyl)stannanes with allylmetals (metal = Si, Ge, Sn) in the presence of BF(3).OEt(2) were carried out. The reactions seem to proceed by the initial formation of an alpha-stannyl-substituted carbocation, which adds to an allylmetal to give the carbocation that is beta to the metal and gamma to tin. The beta-elimination of the metal gives the corresponding allylated product, and the gamma-elimination of tin gives the cyclopropane derivative. In the case of allylsilane, the cyclopropane derivative was formed as a major product, whereas in the case of allylgermane the allylated product was formed predominantly. In the case of the allystannane the allylated product was formed exclusively. These results indicate that the gamma-elimination of tin is faster than the beta-elimination of silicon, but slower than the beta-elimination of germanium and tin. The theoretical studies using ab initio molecular orbital calculations of the carbocation intermediates are consistent with the experimental results. The effect of substituents on silicon was also studied. The introduction of sterically demanding substituents on silicon disfavored the beta elimination of silicon probably because of the retardation of nucleophilic attack on silicon to cleave the carbon-silicon bond. PMID- 10814209 TI - Synthesis of cryptophycin 52 using the sharpless asymmetric dihydroxylation: diol to epoxide transformation optimized for a base-sensitive substrate. AB - A synthesis of cryptophycin 52 (2) is reported using a Sharpless asymmetric dihydroxylation (AD) strategy to install the epoxide moiety. The high stereoselectivity of the AD reaction that allows for an efficient means of preparing the epoxide is in contrast to the standard direct epoxidation of cryptophycin substrates, which proceeds with poor diastereoselectivity. Methodology for conversion of the diol AD product to the requisite epoxide is disclosed. The transformation has been optimized to proceed in high yield in the presence of base sensitive functionality. PMID- 10814210 TI - A stereoselective route to polysubstituted tetrahydroquinolines by benzotriazole promoted condensation of aliphatic aldehydes and aromatic amines AB - By the promotion of benzotriazole (20 mol %), two molecules of anilines (or other arylamines) and two molecules of phenylacetaldehyde (or o bromophenylacetaldehyde) condensed to give a series of 1,2,3,4 tetrahydroquinolines in a stereoselective manner. By the catalysis of SmI(2) or SmI(3), the N-(alpha-aminoalkyl)benzotriazoles derived from anilines and (R) glyceraldehyde acetonide dissociated to the corresponding iminium and enamine species, which underwent asymmetric [4 + 2] cycloadditions to give optically active tetrahydroquinolines. PMID- 10814211 TI - Axially chiral analogues of 4-(Dimethylamino)pyridine: novel catalysts for nonenzymatic enantioselective acylations AB - A concise seven-step synthesis of atropisomeric 3-aryl analogues of DMAP from 4 pyridone 8 has been developed. A representative compound of this class, biaryl (+/-)-15, has been resolved using CSP HPLC and shown to be an efficient nucleophilic catalyst for kinetic resolution of a series of secondary alcohols on both an analytical and preparative scale (stereoselectivity factors, s = 8.9-29). PMID- 10814212 TI - Rational synthesis of meso-substituted chlorin building blocks. AB - Chlorins provide the basis for plant photosynthesis, but synthetic model systems have generally employed porphyrins as surrogates due to the unavailability of suitable chlorin building blocks. We have adapted a route pioneered by Battersby to gain access to chlorins that bear two meso substituents, a geminal dimethyl group to lock in the chlorin hydrogenation level, and no flanking meso and beta substituents. The synthesis involves convergent joining of an Eastern half and a Western half. A 3,3-dimethyl-2,3-dihydrodipyrrin (Western half) was synthesized in four steps from pyrrole-2-carboxaldehyde. A bromodipyrromethane carbinol (Eastern half) was prepared by sequential acylation and bromination of a 5 substituted dipyrromethane followed by reduction. Chlorin formation is achieved by a two-flask process of acid-catalyzed condensation followed by metal-mediated oxidative cyclization. The latter reaction has heretofore been performed with copper templates. Investigation of conditions for this multistep process led to copper-free conditions (zinc acetate, AgIO(3), and piperidine in toluene at 80 degrees C for 2 h). The zinc chlorin was obtained in yields of approximately 10% and could be easily demetalated to give the corresponding free base chlorin. The synthetic process is compatible with a range of meso substituents (p-tolyl, mesityl, pentafluorophenyl, 4-[2-(trimethylsilyl)ethynyl]phenyl, 4-iodophenyl). Altogether four free base and four zinc chlorins have been prepared. The chlorins exhibit typical absorption spectra, fluorescence spectra, and fluorescence quantum yields. The ease of synthetic access, presence of appropriate substituents, and characteristic spectral features make these types of chlorins well suited for incorporation in synthetic model systems. PMID- 10814213 TI - Enantiospecific total synthesis of the sarpagine related indole alkaloids talpinine and talcarpine as well as the improved total synthesis of alstonerine and anhydromacrosalhine-methine via the asymmetric Pictet-Spengler reaction. AB - The enantiospecific total synthesis of talpinine 1 and talcarpine 2 has been accomplished from D-(+)-tryptophan in 13 steps (11 reaction vessels) in 10% and 9.5% overall yields, respectively. Moreover, this synthetic approach has been employed for the improved synthesis of alstonerine 3and anhydromacrosalhine methine 4 in 12% and 14% overall yield, respectively. A convenient synthetic route for the enantiospecific, stereospecific preparation of the key intermediate (-)-N(a)-H, N(b)-benzyl tetracyclic ketone 15a via the asymmetric Pictet-Spengler reaction on a multihundred-gram scale has been developed. A diastereocontrolled (>30:1) anionic oxy-Cope rearrangement and the intramolecular rearrangement to form ring-E and an N(b)-benzyl/N(b)-methyl transfer reaction also served as key steps. This general approach can now be utilized for the synthesis of macroline/sarpagine related indole alkaloids and their antipodes for biological screening. PMID- 10814214 TI - Isolation, structural characterization, and synthesis of a naturally occurring bisfuranopseudopterane ether: biskallolide A. Evidence for a carbocation intermediate during the facile conversion of kallolide A and isokallolide A into various solvolysis products. AB - The West Indian alcyonacean Pseudopterogorgia bipinnata (Verrill, 1864) is shown to contain a novel bisditerpenoid ether: biskallolide A (2). The structural assignment of 2 was mainly based on 1D and 2D NMR and MS spectral data and was further confirmed by synthesis. The 2-C-alkoxylation of furanopseudopteranes kallolide A (1) and isokallolide A (8) occurs spontaneously in some solvents and involves replacement of the C2 hydroxyl with an alkoxyl group to yield solvolysis products that display net retention of configuration. The facile solvolytic 2-C acyloxylation of kallolide A was achieved readily under similar circumstances to afford kallolide A acetate (4) as the sole product. Mechanistic details in conversion of alcohols 1 and 8 into various solvolysis products, including dimeric ethers 2 and 9, were investigated in this study. Solvolysis of kallolide A and isokallolide A in [(18)O]-labeled solvent demonstrated that the C2 alkoxyl of the solvolysis products originated from the solvent, suggesting that these conversions may proceed through an S(N)1 mechanism with generation of a carbocation intermediate. The chemical structures of kallolide A derivatives 3 7and those of isokallolide A congeners 9-11 were established by detailed analysis of the spectral data. PMID- 10814215 TI - Conformational studies by dynamic NMR. 74.(1) stereomutations of the conformational enantiomers in peri-substituted 1-acylnaphthalenes AB - Naphthalenes bearing an acyl and a phenyl group in a peri relationship give rise to a pair of enantiomers in the temperature range where the rotations of the acyl group are slow. Such enantiomers were observed by means of low temperature NMR spectra in chiral environments. The barrier to rotation for the acyl substituents, that causes the interconversion of the enantiomers, was demonstrated to be lower than that for the phenyl group. In an appropriately synthesized derivative it was possible to measure the two barriers that were found equal to 10.4 and 15.9 kcal mol(-)(1), respectively. The barriers for the acyl group rotation increase regularly (from 9.5 to 13.2 kcal mol(-)(1)) with the increasing dimension of the RCO groups (R = Me, Et, Pr(i), Bu(t)). When a bromine atom replaces the phenyl group, the enantiomerization barrier for the corresponding acyl derivatives increases significantly. PMID- 10814216 TI - Conformational studies by dynamic NMR. 73.(1) conformational enantiomers of cyclohexene oxide in the solid state PMID- 10814217 TI - Chiral nonracemic synthesis and reactivity of two new endocyclic enamines in the phenyloxazolopiperidine series PMID- 10814218 TI - Simple asymmetric synthesis of 2H-azirines derived from phosphine oxides. PMID- 10814219 TI - Biomimetic-type synthesis of benzo[a]naphthacenequinones related to pradimicinone. PMID- 10814220 TI - Palladium-catalyzed reaction of propargyl nucleophiles with alpha-sulfonyl alpha,beta-unsaturated ketones: A single-step synthesis of Furo PMID- 10814222 TI - Preparation of a resin-bound arene-ruthenium complex and assessment of its use in enol formate synthesis and olefin cyclopropanation PMID- 10814221 TI - Total synthesis of Amaryllidaceae pyrrolophenanthridinium alkaloids via the Ziegler-Ullmann reaction: tortuosine, criasbetaine, and ungeremine. PMID- 10814223 TI - An efficient, stereospecific synthesis of the dimer-selective retinoid X receptor modulator (2E,4E,6Z)-7-[5,6,7,8- tetrahydro-5,5, 8,8-tetramethyl-2-(n propyloxy)naphthalen-3-yl]-3-methyl octa-2,4, 6-trienoic acid. PMID- 10814225 TI - Diiodosilane: A reagent for mild, efficient conversion of carbamates to ureas via isocyanates PMID- 10814224 TI - Short total synthesis of (+/-)-galbulin and (+/-)-isogalbulin using zirconium chemistry. PMID- 10814226 TI - One-Pot synthesis of N-protected amino trifluoromethyl ketones as hydrated hydrochloride salts via the CsF-catalyzed reactions of (Trifluoromethyl)trimethylsilane with N-protected amino esters PMID- 10814227 TI - Rapid photochemical generation of ubiquinol through a radical pathway: an avenue for probing submillisecond enzyme kinetics. PMID- 10814228 TI - Asymmetric synthesis of (2S,6S)- and meso-(2S,6R)-diaminopimelic acids from enantiopure bis(sulfinimines). PMID- 10814230 TI - Double annulation route to highly substituted and functionalized cis bicyclic and tricyclic delta-lactams and imides PMID- 10814229 TI - An oxidative prins cyclization methodology. PMID- 10814231 TI - N-methylputrescine oxidation during cocaine biosynthesis: study of prochiral methylene hydrogen discrimination using the remote isotope method. AB - [structure: see text] The stereoselectivity of N-methylputrescine (3) oxidation to pyrrolinium ion 4 in Erythroxylum coca during cocaine (1) biosynthesis was studied. The remote isotope method was used to advantage. Each enantiomer of 4 monodeuterated N-methylputrescine served as a precursor for plant feeding. To facilitate mass-spectrometric analysis of products, a 2H3 13C-methyl group was also incorporated into the 4-deuterio-N-methylputrescines. Oxidative deamination of N-methylputrescine was found to be stereoselective; the pro-S hydrogen atom is removed with 6-10:1 selectivity. PMID- 10814232 TI - Synthesis and pharmacology of seleninic acid analogues of the inhibitory neurotransmitter gamma-aminobutyric acid. AB - [structure: see text] The first seleninic acid analogues of gamma-aminobutyric acid (GABA) and of the specific GABA(A) agonist piperidine-4-carboxylic acid (isonipecotic acid), 1 and 2, respectively, have been synthesized and shown to be potent agonists at the GABA receptors. PMID- 10814233 TI - Direct synthesis of beta-aminoketones from amides via novel sequential nucleophilic substitution/Michael reaction AB - [reaction: see text] The synthesis of beta-aminoketones from amides can be achieved in a process consisting of sequential nucleophilic substitution at the carbonyl group by vinylmagnesium bromide followed by Michael reaction after quench of the first reaction by water. PMID- 10814234 TI - Long-range through-bond photoactivated sigma bond cleavage in steroids. Intramolecular sensitized debromination AB - [structure: see text] The photolysis of 17alpha-bromo-3alpha-(triphenylsilyloxy) 5alpha-androstane (2; 3alphaTPSO/17alphaBr) and 17alpha-bromo-3alpha (triphenysilyloxy)-5-androstan-6-one (3; 3alphaTPSO/6ketone/17alphaBr) is described. Irradiation of 2 with 266 nm light leads to debromination via intramolecular transfer of triplet excitation energy with a quantum efficiency of 0.0011. Photolysis of 3 with both 266 and 308 nm light leads to debromination with quantum efficiencies of ca. 0.0066. The debromination of 3 is attributed to activation via the ketone excited singlet state, with singlet energy transfer from C6 to C17 ca. 35% efficient and occurring with a rate constant of 1.4 x 10(8) s(-1). PMID- 10814235 TI - Novel hydrazino-carbonyl-amino-methylated polystyrene (HCAM) resin methodology for the synthesis of P1-aldehyde protease inhibitor candidates. AB - [structure: see text] A new strategy for the synthesis of peptidyl and peptidomimetic P1-aldehydes 3 on HCAM solid support is described. The appropriate C-terminal aldehyde precursors were prepared and anchored to a resin support via a semicarbazone linkage (HCAM resin). After synthetic elaboration, acidic hydrolysis efficiently delivered C-terminal target aldehydes 3a-h in good overall yields and in excellent purity. PMID- 10814236 TI - Nalpha-2-mercaptobenzylamine-assisted chemical ligation. AB - [reaction: see text] The selective formation of an amide bond in the presence of unprotected functional groups is a challenging problem of peptide chemistry. A 2 mercaptobenzyl group tethered at the N-terminus of model peptides was observed to facilitate amide bond formation when a peptide thioester was added under mild aqueous conditions. PMID- 10814237 TI - Synthesis of a model for C7-C13 of lankamycin. AB - [structure: see text] A convenient method is reported for construction of the C7 C13 segment of the macrolide antiobiotic lankamycin. PMID- 10814238 TI - Self-organization of spheroidal molecular assemblies in polar solvents AB - [reaction: see text] We report a number of 1:1 noncovalent complexes composed of a symmetrical trisphosphonate and various symmetrical trisammonium or trisamidinium compounds. The spheroidal complexes show high thermodynamic stability, with association constants Ka reaching 10(6) M(-1) in methanol and in some cases even exceeding 10(3) M(-1) in water. The observed Ka values correlate well with the different degree of preorganization of the complexation partners. PMID- 10814239 TI - Outer-ring stereochemical modulation of cytotoxicity in cephalostatins. AB - [structure: see text] 20- and 25'-epimers of cephalostatin 7, prepared by directed unsymmetrical pyrazine synthesis, address outer-ring topographical and stability questions and intimate an oxacarbenium ion rationale for the role in bioactivity of the spiroketal (E/F, E'/F') rings of this class of antitumor agents. PMID- 10814240 TI - Synthesis of multiply substituted, ion channel forming octi(p-phenylene)s: theme and variations. AB - [reaction: see text] To facilitate the access to unique models for biological processes, we examined six different synthetic routes to octi(p-phenylene) rods with lateral and terminal substituents R(L) and R(T). This systematic study allowed us to increase to overall yield for the synthesis of a new class of oligo(p-phenylene) ionophores about 20 times and to provide general insights into the practicability of synthetic routes to multiply substituted molecular rods. PMID- 10814241 TI - F8BINOL, an electronically perturbed version of BINOL with remarkable configurational stability AB - [structure: see text] Substitution of hydrogens by fluorines at the 5, 5', 6, 6', 7, 7', 8, and 8' positions of BINOL strongly affects distribution of electron density within the biaryl skeleton but has a very small influence on the torsion angle. The most important consequence of this structural alteration is the dramatic increase in configurational stability of homochiral F8BINOL. PMID- 10814242 TI - TiCl4-mediated reduction of 1,3-diketones with BH3-pyridine complex: a highly diastereoselective method for the synthesis of syn-1,3-Diols AB - [reaction: see text] 1,3-Diketones can be reduced in high yields and with excellent diastereoselectivity to the corresponding syn-1,3-diols by carrying out the reaction with BH3-pyridine complex in CH2Cl2 at -78 degrees C in the presence of an equivalent of TiCl4 and 0.1 equiv of pyridine. This protocol shows a general character: excellent results are obtained when the groups bound to the carbonylic functions are linear or branched carbon chains and aromatic or benzylic frameworks as well. PMID- 10814243 TI - Remarkably high asymmetric amplification in the chiral lanthanide complex catalyzed hetero-Diels-Alder reaction: first example of the nonlinear effect in ML3 system AB - [reaction: see text] A remarkably high asymmetric amplification was realized in the Yb[(R)-BNP]3-catalyzed hetero-Diels-Alder reaction as the first example in the metal/chiral ligand 1:3 system. The mechanism may be explained by the autogenetic formation of the enantiopure complex as the most active catalyst. The enantiomer-discriminative formation of homochiral ML3 complexes is quite general within the lanthanide metal ions with similar ionic radii to that of the ytterbium ion. PMID- 10814244 TI - Stereoselective synthesis of functionalized trans-2,5-disubstituted tetrahydrofurans AB - [reaction: see text] The addition of the titanium enolates of N-acetyl, N propionyl, and N-bromoacetyl (R)-oxazolidin-2-ones to gamma-lactol 2, derived from (S)-glutamic acid, afforded trans- and cis-2,5-disubstituted tetrahydrofurans (trans/cis ratio: R = H, 2:1; R = Me, 8:1; R = Br, 10:1) after desilylation with aqueous HF/CH3CN. Chromatographic separation and LiBH4 reduction allowed the efficient preparation of the corresponding trans-2,5 disubstituted tetrahydrofuran diols and the recovery of the chiral auxiliary. PMID- 10814245 TI - Diastereoselective formation of cyanohydrins from alpha-alkoxy aldehydes AB - [reaction: see text] The reaction of alpha-alkoxy aldehydes with Et4NAg(CN)2 or Me3SiCN in the presence of MgBr2 x OEt2 in CH2Cl2 at 0 degrees C gives the corresponding syn cyanohydrins in good yield with high diastereoselectivity. Excess MgBr2 x OEt2 (typically 5 equiv) is required for high diastereoselectivity. Et4NAg(CN)2 (but not Me3SiCN) is sufficiently reactive to give cyanohydrins at -78 degrees C, and higher diastereoselectivity is obtained at this temperature. PMID- 10814246 TI - Tribenzo AB - [structure: see text] Dibenzylammonium (DBA+) ions thread through the cavity of tribenzo[27]crown-9 (TB27C9) to generate [2]pseudorotaxanes that are stabilized principally by hydrogen-bonding interactions. The stabilities and complexation kinetics associated with these pseudorotaxanes depend markedly on the nature of the substituents situated on the phenyl rings of the DBA+ ions. For example, the complex formed between TB27C9 and the DBA+ ion bearing electron-withdrawing pCO2Me substituents is stronger than that obtained from TB27C9 and the "parent", unsubstituted DBA+ ion itself. Furthermore, the "parent" complex equilibrates much more rapidly with its uncomplexed components than do the complexes generated from TB27C9 and substituted DBA+ ions. PMID- 10814247 TI - An efficient and stereoselective synthesis of xerulin via Pd-catalyzed cross coupling and lactonization featuring (E)-lodobromoethylene as a novel two-carbon synthon. AB - [structure: see text] Xerulin, an inhibitor of cholesterol biosynthesis, has been synthesized from commercially available (E)-1-bromopropene, acetylene, and propynoic acid in five steps (longest linear sequence) in 30% overall yield and >96% stereoselectivity. The preparation of (E)-iodobromoethylene and its use in the Pd-catalyzed cross coupling are two of the novel aspects of the synthesis reported herein. PMID- 10814248 TI - Palladium-catalyzed arylation of alpha-methylene-gamma-butyrolactone: 3 benzylfuran-2(5H)-ones vs (Z)-benzylidene-gamma-butyrolactones and their reduction to 3-benzyl-gamma-butyrolactones AB - [reaction: see text] The palladium-catalyzed arylation of the alpha-methylene gamma-butyrolactone proceeds in good yields and may be directed toward the synthesis of 3-benzylfuran-2(5H)-ones when the starting aryl iodides contain strongly electron-withdrawing groups. The combined palladium-catalyzed arylation/hydrogenation of the alpha-methylene-gamma-butyrolactone represents a new simple entry into functionalized alpha-benzyl-gamma-butyrolactones. PMID- 10814249 TI - Sequential Diels-Alder reaction of in situ generated 2,3-dimethylenepyrrole and carbodienophiles: rapid synthesis of 2,3,6,7-tetrasubstituted carbazoles AB - [reaction: see text] 2,3,6,7-Tetrasubstituted-1,2,3,4,5,6,7,8-octahydrocarbazoles 2 were synthesized in 46-90% yields by sequential Diels-Alder reactions from N benzyl-2,5-dimethyl-3,4-bisacetoxymethylpyrrole (1) and dienophiles such as maleic anhydride, maleimides, ethyl maleate, fumaronitrile, and ethyl acrylate. The 2,3,6,7-tetrasubstituted carbazoles 3 were then synthesized in 32-87% yields from 2 by oxidation with DDQ. PMID- 10814250 TI - Competition between cyclization, cleavage, and reverse hydrogen transfer in 1,4 hydroxybiradicals: crystal structure--solid state reactivity correlations AB - [reaction: see text] To provide experimental evidence on the relationship between stucture and reactivity in 1,4-hydroxybiradical intermediates, the Norrish type II photochemistry of a homologous series of spirobenzoyladamantane derivatives was investigated in the crystalline state and the outcome correlated with the structures of the compounds as determined by X-ray crystallography. The results provide an unusually detailed and compelling picture of the geometric factors responsible for the partitioning of these reactive intermediates among cleavage, cyclization, and reverse hydrogen transfer. PMID- 10814251 TI - Replacement of the phosphorodiester linkages of RNA with guanidinium linkages: the solid-phase synthesis of ribonucleic guanidine. AB - [reaction: see text] Replacement of the negatively charged phosphodiester linkages of RNA with positively charged guanidinium linkages provides the polycationic ribonucleic guanidine (RNG). RNG is anticipated to bind strongly to target DNA/RNA through the specific interactions of nucleobases and the attractive electrostatic interactions of backbones. Preparation of building blocks and the solid-phase synthesis of RNG are reported. Both trimeric and pentameric uridyl RNG have been synthesized. PMID- 10814252 TI - A reiterative approach to 2,3-disubstituted naphthalenes and anthracenes AB - [reaction: see text] Simple bis(bromoethynyl)arenediynes are easily prepared by the desilylative halogenation of the corresponding trimethylsilyl derivatives. Cycloaromatization of these halogenated enediynes leads to the otherwise difficult to prepare 2,3-dibromoarenes in good yield. Alkynylation of the resulting haloaromatic compound regenerates the soluble enediyne system, homologated by one aromatic ring. This iterative methodology can be terminated by the cycloaromatization of the unsubstituted enediyne, providing the simple acene hydrocarbon. PMID- 10814253 TI - Facile solid-phase construction of indole derivatives based on a traceless, activating sulfonyl linker AB - [reaction: see text] Palladium-mediated coupling/intramolecular indole cyclization of terminal alkynes with resin 8, followed by cleavage of the sulfonamide linkage, were executed under mild conditions to provide diverse indoles 10 in excellent yield and purity. This chemistry benefits from a dual activation process that derives from use of a traceless N-sulfonyl linker. Also, direct mercuration of 9 (X = H, R = 4-Me-C6H4), followed by palladium-mediated coupling with methyl acrylate, efficiently provided 3-functionalized product 12. PMID- 10814255 TI - A Biomimetic Synthesis of the Pauciflorine A and B Skeleton. PMID- 10814254 TI - A concise enantioselective entry to the synthesis of deoxy-azasugars. AB - [reaction: see text] A concise enantioselective preparation of oxazolidinylpiperidine 4, a key intermediate in the synthesis of glycosidase inhibitors such as 1-deoxymannojirimycin or 1-deoxygalactostatin, has been developed. Sharpless catalytic asymmetric epoxidation of (E)-2,4-pentadienol followed by treatment with allyl isocyanate afforded epoxy carbamate 8. Regioselective intramolecular ring opening promoted by sodium bis(trimethylsilyl)amide and ring-closing metathesis provided the bicyclic intermediate 4 in high enantiomeric purity. The four-step sequence takes place in 51% overall yield. PMID- 10814257 TI - Rearrangement and degradation of cephalosporins and penicillins in the presence of mercury(II) trifluoroacetate AB - [reaction: see text] Cephalosporins and penicillins rearrange under the influence of mercury(II) trifluoroacetate in methanol to non-beta-lactam products. The mechanisms of the rearrangements are different in the two cases. Whereas the open chain aminoacrylic acid derivative 4 is produced from cephalosporins, the oxazole 7 and the propionamide 6 derivatives are the products from penicillins. PMID- 10814256 TI - Total syntheses of (-)-haemanthidine, (+)-pretazettine, and (+)-tazettine. AB - [structures: see text] The total syntheses of the amaryllidaceae alkaloids haemanthidine, pretazettine, and tazettine as optically pure enantiomers are reported. Using D-mannose as the starting material, the critical relative stereochemical relationships are established with an intramolecular nitrone alkene cycloaddition reaction. The synthetic route leads successively to (-) haemanthidine and then to (+)-pretazettine and (+)-tazettine, taking advantage of the well-established complex relationships among these three alkaloids. PMID- 10814258 TI - Use of gamma-carboxy-alpha,beta-unsaturated aldehydes as synthetic equivalents of beta,gamma-unsaturated aldehydes in a novel stereoselective approach to diketides AB - [reactions: see text] A two-step aldol/Pd-catalyzed formate reduction sequence allows for convenient, stereoselective access to products of great potential in synthetic approaches to polyketides. A novel diastereoselective Pd-catalyzed formate reduction of allylic acetates is reported. PMID- 10814259 TI - Synthesis of porphyrins bearing trans-thiols AB - [structure: see text] A route to porphyrins bearing trans-thiols is described using a thioacetyl-containing aldehyde or a thioacetyl-containing dipyrromethane in the presence of catalytic BF3 x OEt2 followed by oxidation. Metal complexation and ammonium hydroxide induced acetyl removal provides a route to these important molecular systems for future electronics experiments in which the thiols would serve as the adhesion points to gold probes. PMID- 10814261 TI - Chirally modified zeolites as reaction media: photochemistry of an achiral tropolone ether AB - [structure: see text] Zeolites modified with chiral inductors serve as media for performing chiral induction during photochemical reactions of organic molecules. The photochemical behavior of achiral tropolone ethyl phenyl ether illustrates this unique feature of a zeolite. PMID- 10814260 TI - A versatile approach to PI(3,4)P2, PI(4,5)P2, and PI(3,4,5)P3 from L-(-) quebrachitol. AB - [reaction: see text] A versatile synthesis of PI(3,4)P2, PI(4,5)P2, and PI(3,4,5)P3 is disclosed, starting from L-(-)-quebrachitol, a byproduct of latex production. The crystalline nature of most intermediates and the utilization of inexpensive protecting groups facilitate this synthetic route and its scale-up. PMID- 10814262 TI - A short and efficient stereoselective synthesis of the polyhydroxylated macrolactone (+)-aspicillin AB - [structures: see text] A short and efficient synthesis of the polyhydroxylated macrolactone (+)-aspicilin 1 using a stereoselective lithium perchlorate mediated addition of allyltributyltin to the equatorially disposed carboxaldehyde of 3 (derived from (R',R',R,S) butane diacetal protected butane tetrol 2) as the key step is described. Terminal group manipulation and Masamune-Roush olefination using phosphonate ester 4 followed by macrocyclization via ring closing metathesis afforded the natural product after partial hydrogenation and global deprotection. PMID- 10814263 TI - Vesicles accelerate proton transfer from carbon up to 850-fold. AB - [reaction: see text] We have analyzed the different catalytic effects of surfactant aggregates upon the rate-determining hydroxide ion induced deprotonation reaction of 1. Vesicles are more effective catalysts than micelles, most likely providing a more apolar microenvironment at the substrate binding sites. We suggest that this leads to a catalytic reaction involving less strongly hydrated hydroxide ions. In the case of DODAB and DODAC vesicles, binding of cholesterol to the bilayer further increases the catalytic efficiency. PMID- 10814264 TI - An efficient synthetic approach to highly conjugated porphyrin-based assemblies containing a bipyridine moiety AB - [structure: see text] An efficient and potential stepwise strategy involving the mixed sequence of Stille and Wittig-Horner reactions was used for the preparation of a polyene-substituted bis-porphyrin incorporating a bipyridine moiety. PMID- 10814265 TI - Helical twist sense bias in oligo(phenylene ethynylene)s induced by an optically active flexible tether AB - [graph and structure: see text] A series of tethered phenylene ethynylene oligomers, which undergo a solvent-dependent conformational transition from a random coil to a helix, has been synthesized. The use of trimethylsilyl ether protecting groups on the (+)-tartaric acid-derived tether results in the formation of helices with a large twist sense bias. In contrast, an isopropylidene ketal protecting group or no protecting group is not only ineffective at helical discrimination but may even inhibit helix formation. PMID- 10814266 TI - Lithium ion-selective binding properties of a conformationally constrained tris(spirotetrahydrofuran) secured to an inositol orthoformate platform AB - [structure: see text] The C3V-symmetric triaxially locked trispirotetrahydrofuran 2 binds Li+ ions strongly and Na+ ions much less effectively. The observed discrimination factor is attributed to greater structural preorganiztion and lower solvation prior to selective complexation, and particularly to the formation of a 2:1 sandwich complex involving lithium. 13C NMR studies have defined a reluctance to form a 1:1 species when excess Li+ is present. PMID- 10814267 TI - Synthesis and crystal structure of a unique linear homoditopic ligand bifacially complexed to lithium picrate AB - [structure: see text] Preparation of the first rigid homoditopic tris(tetrahydrofuranyl) ligand with a pair of exotopic binding sites specifically tailored for effective coordination to lithium ions is described. The crystallographically defined structural parameters of the monomeric building block and a 1:2 complex of the dimer with lithium picrate reveal the unique features of these materials. PMID- 10814268 TI - Palladium-mediated ring opening of hydroxycyclopropanes AB - [reaction: see text] The palladium-mediated ring opening of substituted cyclopropanols has been found to take place predominantly at the less substituted C-C bond. Thus, sequential application of the titanium-mediated cyclopropanation of esters and the palladium-mediated ring opening of the resulting cyclopropanols provides a convenient method for functionalizing monosubstituted olefins. PMID- 10814269 TI - Synthesis and evaluation of aminocyclopentitol inhibitors of beta-glucosidases. AB - [reaction: see text] (1R,2S,3S,4R,5R)-4-Amino-5-(hydroxymethyl)cyclopentane-1,2,3 triol 1, prepared from D-glucose, inhibits beta-glucosidases from Caldocellum saccharolyticum (Ki = 1.8 x 10(-7) M) and from almonds (Ki = 3.4 x 10(-6) M). Inhibition is not influenced by N-ethylation (--> 15) but is strongly reduced upon N-acetylation (--> 12). Inversion of stereochemistry at C(5) (--> 14) has little effect on inhibition of beta-glucosidases. These experiments suggest that 1 acts as an analogue of a protonated beta-glucoside. PMID- 10814270 TI - Enantioselective syntheses of 2-alkyl- and 2,6-dialkylpiperidine alkaloids: preparations of the hydrochlorides of (-)-coniine, (-)-solenopsin A, and (-) dihydropinidine. AB - [reaction: see text] Sequences of lithiation-substitution, enantioselective hydrogenation, and diastereoselective lithiation-substitution provide efficient highly enantioselective syntheses of 2-substituted and cis and trans 2,6 disubstituted piperidines. The methodology is demonstrated by syntheses of (-) coniine, (-)-solenopsin A, and (-)-dihydropinidine as their hydrochlorides. PMID- 10814271 TI - Ytterbium(III) triflate/TMSCI: efficient catalyst for imino ene reaction AB - [reaction: see text] Ytterbium trifluoromethanesulfonate [Yb(OTf)3] catalyzed the imino ene reaction of N-tosyl aldimine with alpha-methylstyrene to give a homoallylamine in moderate yield. Furthermore, addition of a catalytic amount of chlorotrimethylsilane (TMSCI) dramatically enhanced the imino ene reaction. PMID- 10814272 TI - Highly efficient, enantioselective synthesis of (+)-grandisol from a C2-symmetric bis(alpha,beta-butenolide). AB - [reaction: see text] A new, very efficient, enantioselective synthesis of the sexual attracting insect pheromone (+)-grandisol has been developed, in which the key step is the double [2 + 2] photocycloaddition of ethylene to a bis(alpha,beta butenolide) readily available from D-mannitol. The C2 symmetry of the substrate and the appropriate protection of the central diol unit are the crucial features for the high diastereofacial discrimination during the cycloaddition process. PMID- 10814273 TI - Total synthesis of both methyl 4a-carba-D-arabinofuranosides. AB - [reaction: see text] The total synthesis of methyl 4a-carba-alpha-D arabinofuranoside (1) and methyl 4a-carba-beta-D-arabinofuranoside (2) has been achieved starting from D-mannose (5). Key transformations included a ring-closing metathesis of diene 11 employing Schrock's catalyst followed by a stereoselective hydrogenation with Wilkinson's catalyst. PMID- 10814274 TI - Triamino-s-triazine triradical trications. An experimental study of triazine as a magnetic coupling unit AB - [structure: see text] Triamino-s-triazine derivatives 3a, 4, and 5 have been prepared, and their cationic states have been analyzed electrochemically. At 298 K, 3a+ has a limited lifetime in CH2Cl2 solution. However, 4+ and 5+ are long lived under such conditions, and quartet states of 4(3+) and 5(3+) are observed by ESR spectroscopy. Variable-temperature ESR analysis and NMR shift susceptibility measurements indicate that 5(3+) is a doublet ground state with a populated quartet state. PMID- 10814275 TI - Enantioselective synthesis of (S)-(+)-pantolactone AB - [reaction: see text] The Prins reaction of a chiral alkylidene morpholinone derived from (1R,2S)-ephedrine and 3-methyl-2-oxobutanoic acid proceeds with good diastereoselectivity to generate a spiro bis-acetal. Lewis acid mediated diastereoselective reductive cleavage of the spiro acetal and subsequent removal of the ephedrine portion generates a alpha-hydroxy-gamma-methoxy carboxamide which is readily converted to (S)-(+)-pantolactone with high enantiomeric excess. PMID- 10814276 TI - Axially chiral amidinium ions as inducers of enantioselectivity in Diels-Alder reactions AB - [reaction: see text] Enantioselective catalysis of Diels-Alder reactions is mostly achieved by coordinating the dienophile to relatively strong chiral Lewis acids. Here we report on a novel approach employing the hydrogen-bond-mediated association of dienophiles to chiral host molecules. In a reaction forming the steroid skeleton of norgestrel, chiral amidinium ions induce 5:ent-5 ratios of up to 2.5:1. Improved and simplified amidinium catalysts may become interesting candidates to perform stereoselective transformations. PMID- 10814277 TI - A novel photochemical vinylcyclopropane rearrangement yielding 6,7-dihydro-5H benzocycloheptene derivatives AB - [reaction: see text] 1-Substituted 1-(2,2-diphenylvinyl)cyclopropanes with electron-accepting groups at C1 undergo a novel rearrangement to benzocycloheptenes on triplet-sensitized irradiation. In some instances competition between rearrangement to cyclopentenes and formation of cycloheptenes takes place. When electron-acceptor groups are not present at C1, conventional ring opening to cyclopentenes occurs exclusively. A mechanism involving intramolecular SET from the diphenylvinyl unit to the electron-acceptor group is proposed for this novel photoreaction. PMID- 10814278 TI - First synthesis of tetrapyrrolylporphyrin AB - [reaction: see text] N-alkyl-substituted meso-tetrapyrrolylporphyrin (TPyrP) and its derivatives were synthesized for the first time via an acid-catalyzed condensation between N-alkyl-2,4-diformylpyrrole and unsubstituted pyrrole and a subsequent deformylation reaction. X-ray structural analysis of formyl substituted TPyrP shows the tilting of meso pyrrole rings ca. 60 degrees to the porphyrin plane. Formyl groups of meso pyrrole rings were removed by treatment with trifluoroacetic acid (TFA) in pyrrole. PMID- 10814279 TI - Total synthesis of (-)-reveromycin B. AB - [structure: see text] The total synthesis of the epidermal growth factor inhibitor reveromycin B (2) is described. A novel, convergent, and stereoselective reaction sequence was utilized to construct the 5,6-spiroketal system 10 which was converted into the natural product 2 by a 16-step sequence. PMID- 10814280 TI - "Reactive filtration": use of functionalized porous polymer monoliths as scavengers in solution-phase synthesis. AB - [reaction: see text] Solid functionalized porous monolithic disks with reactive polymer chains grafted to their inner pore surface have been developed for scavenging excess reagents from reaction mixtures. A poly(chloromethylstyrene-co divinylbenzene) monolith was cut into disks and activated by graft polymerizing 4 vinyl-2,2-dimethylazlactone to its pore surface. In contrast to the direct copolymerization of reactive monomers, grafting increases the accessibility of the reactive groups. Application of the reactive disks is demonstrated in the scavenging of excess amines from reaction mixtures in different solvents. PMID- 10814281 TI - Ligand tuning in asymmetric catalysis: mono- and bis-phospholanes for a prototypical Pd-catalyzed asymmetric allylation reaction AB - [structures: see text] Enantioselectivities and yields comparable to the best catalysts reported previously can be achieved in the addition of potassium dimethyl malonate to diphenylallyl acetate by the use of Pd(0) complexes of bis phospholanes prepared from D-mannitol. By appropriate changes in the C2-C5 substituents, rare example of a useful monophosphine can also be prepared by a similar route. In both instances chirality of C3 and C5 oxygen seems to play a crucial role in the asymmetric induction. PMID- 10814282 TI - Novel palladium(II) complex containing a chelating anionic N-O ligand: efficient carbonylation catalyst AB - [reaction: see text] A novel palladium(II) complex containing chelating anionic pyridine-2-carboxylato and labile tosylato ligands is a highly efficient catalyst for the carbonylation of organic alcohols and olefins to carboxylic acids/esters. Carbonylation of primary, secondary, and tertiary alcohols as well as linear and functionalized terminal olefins was studied. In all cases remarkable activity and selectivity were observed. The catalyst is stable under reaction conditions even in the absence of excess phosphine ligands. PMID- 10814283 TI - Enantioselective synthesis of the [6,6] spiroketal core of reveromycin A. AB - [structure: see text] Reveromycin A (1) belongs to a family of microbial polyketides with unusual structural features and biological activities. The structure of 1 is composed of a [6,6] spiroketal core decorated with highly unsaturated side chains. As a prelude to the synthesis of 1, we present herein a short, efficient, and enantioselective synthesis of the C9-C21 fragment 5 (spiroketal core) of reveromycin A. PMID- 10814284 TI - On the mechanism of palladium-catalyzed coupling of haloaryls to biaryls in water with zinc AB - [reaction: see text] Kinetics and process parameters of coupling and hydro dehalogenation reactions of chloroaryls are studied in the presence of zinc, water, and catalytic Pd/C. Good yields are obtained for the coupling of chlorobenzene, 4-chlorotoluene, and 4-chloro-1,1,1-trifluorotoluene. It is shown that water is actually one of the reagents, reacting with zinc in the presence of palladium to give zinc oxide and hydrogen gas, which then regenerates the Pd0 catalyst for the coupling reaction. PMID- 10814286 TI - Synthesis of 2-anilinotropones via palladium-catalyzed amination of 2 triflatotropone AB - [reaction: see text] A series of 2-anilinotropones were synthesized via the palladium-catalyzed cross coupling of 2-triflatotropone and a wide variety of anilines, sterically hindered as well as electron-poor ones. PMID- 10814285 TI - Synthesis of 2-C-methyl-D-erythritol 4-phosphate: the first pathway-specific intermediate in the methylerythritol phosphate route to isoprenoids. AB - [reaction: see text] 2-C-methyl-D-erythritol 4-phosphate (4), formed from 1-deoxy D-xylulose 5-phosphate (3), is the first pathway-specific intermediate in the methylerythritol phosphate route for the biosynthesis of isoprenoid compounds in bacteria, algae, and plant chloroplasts. In this report, 4 was synthesized from 1,2-propanediol (7) in seven steps with an overall yield of 32% and in an enantiomeric excess of 78%. PMID- 10814287 TI - Stereoselection in the prins-pinacol synthesis of 2,2-disubstituted 4 acyltetrahydrofurans. Enantioselective synthesis of (-)-citreoviral. AB - [reaction: see text] The condensation of allylic diols with unsymmetrical ketones proceeds with high stereoselection to form 2,2-disubstituted 4 acyltetrahydrofurans when the alpha-substituents of the ketone differ substantially in size. A Prins-pinacol condensation of this type is the central strategic step in an enantioselective synthesis of (-)-citreoviral. PMID- 10814288 TI - Regioselective synthesis of beta-N1- and beta-N3-alloxazine nucleosides AB - [structures: see text] A regio- and stereoselective glycosylation of ribose tetraester with persilylated alloxazine to give either beta-N1 or beta-N3 nucleosides is described. The N3 product is potentially of interest as a fluorescent nucleoside and is predicted to have the hydrogen-bonding characteristics of thymidine. PMID- 10814289 TI - A highly efficient synthesis of the hemibrevetoxin B ring system. AB - [reaction: see text] This Letter describes the synthesis of the hemibrevetoxin B tetracyclic ring system in 14 linear transformations from the Danishefsky Kitahara diene. PMID- 10814290 TI - Heteroannulation through copper catalysis: a novel cyclization leading to a highly regio- and stereoselective synthesis of 2-substituted benzothiazolines AB - [reaction: see text] 3-(2-Aminophenylthio)prop-1-yne 8 reacted with aryl iodides 9-18 under palladium-copper catalysis leading to the disubstituted alkynes 19-28 which after tosylation underwent a novel cyclization under copper catalysis to 2 substituted benzothiazolines 29-38. The expected 3-(arylidine)-2,3 dihydrobenzothiazines were not obtained. PMID- 10814291 TI - Synthesis and structure of intramolecularly hydrogen bonded dendrons AB - [reaction: see text] A convenient synthesis of monodendrons whose conformation is restricted through the intervention of intramolecular hydrogen bonding and repulsive electrostatic interactions is described. X-ray crystal structure analysis of the second generation dendron shows the presence of a propeller-type secondary structure and indicates that the dendrons have assembled into a symmetrically interdigitated dimer in the solid state. 1H NMR and IR spectral data are in agreement with the presence of intramolecular hydrogen bonding between the amides and the pyridine N throughout the dendron structure in solution. PMID- 10814293 TI - Regioselective bromination of organic substrates by tetrabutylammonium bromide promoted by V2O5-H2O2: an environmentally favorable synthetic protocol AB - [reaction: see text] Vanadium pentoxide very effectively promotes the bromination of organic substrates, including selective bromination of some aromatics, by tetrabutylammonium bromide in the presence of hydrogen peroxide; mild conditions, high selectivity, yield, and reaction rate, and redundancy of bromine and hydrobromic acid are some of the major advantages of the synthetic protocol. PMID- 10814294 TI - A new procedure for the preparation of beta-keto-delta-lactones from sugars and their transformation into glycosyl acceptors in disaccharides synthesis AB - [reaction: see text] Glycals are effective starting materials for the synthesis of enantiopure beta-ketone-delta-lactones. They are easily transformed, through a two-step, one-pot reaction, into the corresponding alpha,alpha'-dioxothiones which in turn can be quantitatively trapped with dienophiles in inverse electron demand [4 + 2] cycloadditions. The reaction of dioxothione 8b with endo and exo glucals allowed the elaboration of a new protocol to prepare 2-thio- or 2 deoxydisaccharides stereoselectively. PMID- 10814292 TI - Allyl group as a protecting group for internucleotide phosphate and thiophosphate linkages in oligonucleotide synthesis: facile oxidation and deprotection conditions. AB - [reaction: see text] The allyl group, which serves as a protecting group for an internucleotide bond for both phosphates and phosphorothioates, can be easily removed by good nucleophiles under weakly basic or neutral conditions. For a practical synthesis on solid support, camphorsulfonyloxaziridine was used as the oxidizing agent for synthesizing DNA, while the Beaucage reagent was used for preparing phosphorothioate oligomers. Both types of oligonucleotides were easily deprotected by concentrated ammonium hydroxide containing 2% mercaptoethanol. PMID- 10814295 TI - Alkylation of aromatic aldehydes with boron halide derivatives AB - [reaction: see text] Aryl aldehydes react with alkylboron chloride derivatives in the presence of base to generate the corresponding arylalkylmethanols in good yields. A variety of alkylboron chlorides can be utilized in the new alkylation reaction. PMID- 10814296 TI - Antimitotic diterpenes from Erythropodium caribaeorum test pharmacophore models for microtubule stabilization. AB - [structure: see text] Six new antimitotic diterpenes, 2-7, have been isolated from the Caribbean octocoral Erythropodium caribaeorum. Structural variations encountered in this group of natural products test recently proposed pharmacophore models for microtubule stabilizing compounds. PMID- 10814297 TI - A ROMPGEL-supported N-hydroxysuccinimide: a host of acylations with minimal purification AB - [reaction: see text] A novel N-hydroxysuccinimide ring-opening metathesis polymer is described as a recyclable supported acyl transfer reagent. Amides, carbamates, ureas, Weinreb amides, and hydroxamic acids are all obtained in excellent yields and purities from amines with minimal purification. PMID- 10814298 TI - The first O-NO bond energy scale in solution: heterolytic and homolytic cleavage enthalpies of O-nitrosyl carboxylate compounds AB - [reaction: see text] The first series of O-NO bond dissociation enthalpies was determined in solution for eight O-nitrosyl carboxylate compounds by direct titration calorimetry with a thermodynamic cycle. The derived bond energy data may serve as a quantitative guide to predict the NO binding and releasing abilities of the related amino acids. PMID- 10814299 TI - Correlation of substituent effects and energy levels of the two lowest excited states of the azulenic chromophore AB - [structure: see text] Experimental and calculated data show that the relative energy levels of the two lowest excited states of azulene are sensitive to the nature and position of substituents on the nonalternant hydrocarbon. Extending such investigations led to a rational explanation for some of the baffling data on azulenic bacteriorhodopsin analogues in the literature. PMID- 10814300 TI - New hydrogen-bonded donor-acceptor pairs between dipyridylacetylenes and 2,5 dichloro-3,6-dihydroxyl-1,4-benzoquinone AB - [structure: see text] The crystalline donor-acceptor hydrogen-bonding complexes between 2,5-dichloro-3,6-dihydroxy-1,4-benzoquinone (chloranilic acid) and dipyridylacetylenes (DPA) [2,2'-DPA, 3,3'-DPA, and 4,4'-DPA] were prepared, and crystal structures were revealed by X-ray analysis. The structures of the complexes are formed by intermolecular hydrogen-bonding interactions and demonstrate three supramolecular architectures based on a new common supramolecular synthon, which allows the formation of a different stacking arrangement and ionicity. PMID- 10814301 TI - Synthesis of the spirocyclic cyclohexadienone ring system of the schiarisanrins. AB - [structure: see text] Studies on the synthesis of the spirocyclic cyclohexadienone ring system 2 of the schiarisanrin family of natural products 1 are described and were based on the Lewis acid-promoted C-alkylation of the corresponding phenolic precursor. PMID- 10814302 TI - Synthesis of crossed [2 + 2] photocycloadducts: a novel approach to the synthesis of bridged bicyclic alkenes. AB - [reaction: see text] A solution to the synthesis of "crossed" intramolecular [2 + 2] photocycloadducts has been achieved. Through the use of a temporary heteroatom linker, the equivalent of a crossed photocycloadduct can be accessed at the expensive of the normal "straight" adduct. Selectivity as high as 94:6 for the "crossed" adduct has been observed. PMID- 10814303 TI - Total asymmetric synthesis of the aphidicolin derivative (11R)-(-)-8-epi-11 hydroxyaphidicolin using tandem transannular Diels-Alder/aldol reactions. AB - [reaction: see text] Aphidicolin unnatural derivative (2) was synthesized using a new tandem transannular Diels-Alder/aldol methodology. The 8-epi-aphidicolane skeleton is constructed in a highly diastereoselective manner and converted into (11R)-(-)-8-epi-11-hydroxyaphidicolin (2). An efficient method for the difficult C16 funtionalization is presented. PMID- 10814304 TI - New diffusion-edited NMR experiments to expedite the dereplication of known compounds from natural product mixtures. AB - [reaction: see text] We present two new diffusion-edited NMR experiments, improved DECODES and HETDECODES, that sort the constituents in a mixture by their individual diffusion coefficients. These experiments should allow the partial NMR spectral assignment and cursory structure elucidation of compounds in a complex mixture as an aid in the dereplication of known or nuisance compounds. PMID- 10814305 TI - Chemical synthesis of cross-linked purine nucleosides. AB - [reaction: see text] An efficient route to all of the possible cross-linked 2' deoxypurines 1-3 has been developed by means of the Pd-mediated C-N bond formation in the key step. Utilizing this protocol, the synthesis of the first unnatural protected purine trimeric adduct 4 has been accomplished. PMID- 10814306 TI - A new complex of palladium-thiourea and carbon tetrabromide catalyzed carbonylative annulation of o-hydroxylarylacetylenes: efficient new synthetic technology for the synthesis of 2,3-disubstituted benzo[b]furans. AB - [reaction: see text] A highly effective cocatalysis system (PdI2-thiourea and CBr4) was developed for carbonylative cyclization of both electron-rich and electron-deficient o-hydroxylarylacetylenes to the corresponding methyl benzo[b]furan-3-carboxylates. PMID- 10814307 TI - Design and synthesis of novel conformationally restricted peptide secondary structure mimetics AB - [structure: see text] A facile synthesis of the novel conformationally restricted reverse turn mimetic is described. The key features are the preparation of the alpha-keto amide and tandem bicyclic ring formation. PMID- 10814308 TI - Traceless solid-phase synthesis of chiral 3-aryl beta-amino acid containing peptides using a side-chain-tethered beta-amino acid building block. AB - [reaction: see text] A general method for the attachment of a chiral aromatic side-chain-containing beta-amino acid to a polymer support using a traceless silyl linkage strategy has been developed. Using this building block, solid-phase synthesis was carried out to obtain tripeptide analogues with the aromatic ring either unsubstituted or halogenated (Br, I) at the position of the silyl group. The building blocks could generate libraries of peptidomimetics or cyclic peptides containing beta-amino acids with nonpolar side chains. PMID- 10814309 TI - Aryl radical cyclizations of 1-(2'-bromobenzyl)isoquinolines with AIBN-Bu3SnH: formation of aporphines and indolo AB - [reaction: see text] Radical cyclization of alkoxy-substituted 1-(2'-bromobenzyl) 3,4-dihydroisoquinolines 1 with AIBN-Bu3SnH gave 6a,7-dehydroaporphines 2 preferentially. A steric repulsion between the respective alkoxy groups at the 7- and 3'-positions gave 5,6-dihydroindolo[2,1-a]isoquinolines 3 in a "disfavored" 5 endo cyclization mode. Radical cyclizations of the related substrates, such as 1 (2'-bromobenzoyl)isoquinolines or 1-(2'-bromo-alpha-hydroxybenzyl)isoquinolines, were also found to give the corresponding oxoaporphines or oxyaporphines. PMID- 10814310 TI - Michael addition of selenoamides to alpha,beta-unsaturated carbonyl compounds: stereocontrolled synthesis of delta-oxo selenoamides and their reactivity AB - [reaction: see text] Lithium eneselenolates generated from selenoamides underwent Michael addition to alpha,beta-unsaturated esters and ketones with high diastereoselectivity to give delta-oxo selenoamides in moderate to high yields within a few seconds. Further selective transformations of the delta-oxo selenoamides were also achieved. PMID- 10814311 TI - Chemospecific monofunctionalization of alpha-cyclodextrin in the solid state AB - [reaction: see text] The properties of the self-assembling aziadamantane inclusion complex with two alpha-cyclodextrin molecules have been exploited to perform a chemospecific monofunctionalization of alpha-cyclodextrin. The insertion of the photochemically generated carbenes takes place chemospecifically into the cyclodextrin's C-3-OH and C-2-OH bonds in 39 and 18% yield, respectively. This model reaction surpasses conventional methods in terms of yield as well as selectivity. PMID- 10814312 TI - The total synthesis of pantocin B. AB - [reaction: see text] Pantocin B, an unusual antibiotic produced by Erwinia herbicola, effectively controls E. amylovora, the pathogen causing the plant disease fire blight. A total synthesis of pantocin B from L-alanine, glycine, and L-malic acid is reported. PMID- 10814313 TI - Solid-phase synthesis of N-alkyl-N-(beta-keto)amides and 1,2,4,5-tetrasubstituted imidazoles using a traceless cleavage strategy AB - [reaction: see text] The solid-phase synthesis of N-alkyl-beta-keto)amides and 1,2,4,5-tetrasubstituted imidazoles was demonstrated using a traceless cleavage strategy based on benzylic acylammonium chloride reactivity. The approach enables the assembly of diverse compounds in a minimal number of steps in moderate to excellent yield (23-88%) and high purity (64-100%). PMID- 10814314 TI - Application of olefin cross-metathesis to organometallics. Synthesis of unsaturated ferrocenyl alcohols and ketones AB - [reaction: see text] The application of olefin cross-metathesis to terminally unsaturated alpha-hydroxy- and alpha-ketoferrocenes is reported. Both substrates provided the terminally functionalized olefins in moderate to good yield. These examples represent the first cases of cross-metathesis of substrates containing an organometallic functional group. PMID- 10814315 TI - Oxidative photodecarboxylation of alpha-hydroxycarboxylic acids and phenylacetic acid derivatives with FSM-16 AB - [reaction: see text] FSM-16, a mesoporous silica, was found to catalyze oxidative photodecarboxylation of alpha-hydroxy carboxylic acid and phenyl acetic acid derivatives to afford the corresponding carbonyl compounds. Furthermore, FSM-16 proved to be reuseable by recalcination at 450 degrees C after the reaction. PMID- 10814316 TI - Enantiocontrolled synthesis of trialkyl-substituted stereogenic carbons. A general route to cis-3,5-dialkyl gamma-lactones AB - [reaction: see text] Lewis acid treatment of tertiary Co2(CO)6-propargylic alcohols having a stereochemically defined benzyloxy group at the gamma-benzyl position yielded after cobalt demetalation sec-dialkyl bishomopropargylic alcohols in good yields. The reaction is highly stereoselective and predictable, providing pure stereoisomers. The use of benzyl-alpha,alpha'-d2 ethers permitted the stereoselective d-labeling of methines and methylenes. Very simple chemical manipulations provided a general methodology to obtain the enantiomers of 3,5 dialkyl-gamma-lactones. PMID- 10814317 TI - Steric bulk at position 454 in Saccharomyces cerevisiae lanosterol synthase influences B-ring formation but not deprotonation. AB - [reaction: see text] Lanosterol synthase converts oxidosqualene to the tetracyclic sterol precursor lanosterol. The mutation experiments described here show that an active-site valine residue in lanosterol synthase contributes to cyclization control through steric effects. Mutating to smaller alanine or glycine residues allows formation of the monocyclic achilleol A, whereas the leucine and isoleucine mutants make exclusively lanosterol. The phenylalanine mutant is inactive. PMID- 10814318 TI - Synthesis of 3-ethoxycarbonyl-4-hydroxyquinoline N-oxides from the Baylis-Hillman adducts of o-nitrobenzaldehydes AB - [reaction: see text] The reaction of the Baylis-Hillman adducts la-e of o nitrobenzaldehydes and trifluoroacetic acid at 60-70 degrees C gave 3 ethoxycarbonyl-4-hydroxyquinoline N-oxide derivatives 3a-e in good to moderate yields. PMID- 10814320 TI - Novel synthesis of substituted thiophenes by palladium-catalyzed cycloisomerization of (Z)-2-en-4-yne-1-thiols AB - [reaction: see text] The first example of palladium-catalyzed cycloisomerization of (Z)-2-en-4-yne-1-thiols 1 to give substituted thiophenes 2 is reported. Cycloisomerization reactions are carried out under nitrogen at 25-100 degrees C in N,N-dimethylacetamide as the solvent in the presence of catalytic amounts of PdI2 in conjunction with KI to give the corresponding thiophenes in 43-94% yield. PMID- 10814319 TI - Synthesis and study of a lipophilic alpha-keto amide inhibitor of pancreatic lipase. AB - [reaction: see text] A lipophilic alpha-keto amide, inhibitor of pancreatic lipase, was synthesized using a lipidic 2-amino alcohol as backbone. The chiral key intermediate 2-(tert-butyloxycarbonylamino)-D-undecen-5-ol was synthesized starting from D-glutamic acid. The inhibitor formed a stable monomolecular film at the air/water interface as shown by a force/area curve. Inhibition studies using the monomolecular film technique with mixed films of 1,2-dicaprin containing variable proportions of the inhibitor showed a 50% decrease in lipase activity at a 0.14 molar fraction. PMID- 10814321 TI - Dipolar cycloaddition of rhodium-generated carbonyl ylides with p-quinones. AB - [reaction: see text] The dipolar cycloaddition of carbonyl ylides generated by the rhodium-catalyzed decomposition of delta- and epsilon-carbonyl-alpha diazoketones with p-quinones leads to both C=O and C=C addition products. The product ratio is solvent- and catalyst-dependent and has been optimized to favor formation of either product. The C=C addition products of naphthoquinones are used in the assembly of structures hybridizing the illudin and anthraquinone anticancer agents. PMID- 10814322 TI - Mechanism of oxygen transfer in the epoxidation of an olefin by molecular oxygen in the presence of an aldehyde AB - [reaction: see text] The reaction pathway for peroxide-initiated aldehyde mediated oxidation of olefins to epoxides by molecular oxygen has been studied. The pathways of reaction via a peroxy acid or an acyl peroxy radical have been differentiated by investigation of the reaction of 4 with oxygen to provide 6 via 8. PMID- 10814323 TI - Occurrence of precursors of regular tricyclopolyprenoids in recent sediments AB - [structure: see text] Two novel C24 tricyclic alkenes of the cheilanthane series have been isolated from the solvent extract of a sulfur-rich recent sediment and their structures determined by NMR studies. They represent the first example of functionalized tricyclic hydrocarbons structurally related to the tricyclopolyprenanes widespread in ancient sediments and petroleum identified in recent sediments. PMID- 10814324 TI - Enantiopure eta4-(1-sulfinyldiene)iron(0) tricarbonyl complexes as templates for carbocycle construction via ring-closing metathesis AB - [reaction: see text] Enantiomerically pure 1-(1Z,3E)-sulfinyl iron(0) dienols have been elaborated to bis-olefins capable of undergoing ring-closing metathesis chemistry. Using Grubbs' ruthenium carbene catalyst, a six-membered carbocycle (with one chiral center) and seven-, eight-, and nine-membered carbocycles (with two chiral centers) have been prepared. Novel transformations include the LiClO4 promoted allylation of an alkylidene malonate and a reductive sulfur-carbon bond cleavage to convert a vinyl sulfoxide into an alkene with SmI2. PMID- 10814325 TI - Convenient, in situ generation of anhydrous hydrogen iodide for the preparation of alpha-glycosyl iodides and vicinal iodohydrins and for the catalysis of Ferrier glycosylation AB - [reaction: see text] Anhydrous hydrogen iodide is generated in situ by the reaction of solid iodine and a thiol. The HI thus generated has been employed for the efficient preparation of alpha-glycosyl iodides and vicinal iodohydrins from the corresponding glycosyl acetates and epoxides, respectively, and for Ferrier glycosylation of alcohols and thiols. PMID- 10814327 TI - Acceleration of acid-catalyzed transesterification of 2-hydroxypropyl-p nitrophenyl phosphate by organic solvents AB - [structure: see text] Transesterification of 2-hydroxypropyl-p-nitrophenyl phosphate in the presence of 0.092 M HClO4 is 50-5000 times faster in acetonitrile, 1,4-dioxane, methanol, ethanol, N,N-dimethylformamide, or dimethyl sulfoxide than in water. This demonstrates the importance of tuning the microenvironments in designing synthetic nucleases. PMID- 10814326 TI - Novel synthesis of the ocular age pigment A2-E: new method for substituted pyridine synthesis via azaelectrocyclization. AB - [reaction: see text] The formal synthesis of the ocular age pigment A2-E was achieved by the efficient one-pot preparation of the substituted pyridine, which involves the aza-6pi-electrocyclization of the Schiff base derived from (E)-3 carbonyl-2,4,6-trienal followed by oxidation. PMID- 10814328 TI - Asymmetric synthesis of bicyclic ketones having an angular substituent via Ti(II) alkoxide-mediated tandem cyclization of trisubstituted olefinic substrates AB - [reaction: see text] Angularly substituted, optically active bicyclic ketones of up to 94% ee were prepared by the Ti(II) alkoxide-mediated tandem cyclization of open-chain substrates, that is, 8-phenylmenthyl enynoates having a trisubstituted double bond. PMID- 10814329 TI - Acid-catalyzed cyclization of vinylsilanes bearing an amino group. Stereoselective synthesis of pyrrolidines AB - [reaction: see text] In the presence of an acid catalyst, vinylsilanes 1 bearing an amino group protected by an electron-withdrawing group were smoothly cyclized to 2-(silylmethyl)pyrrolidines 2. This cyclization was utilized for the stereoselective synthesis of 2,n-disubstituted pyrrolidines (n = 3-5). The cyclized products could be converted to the corresponding alcohols by oxidative cleavage of the carbon-silicon bond with TBAF and H2O2. PMID- 10814330 TI - Lactone synthesis based on atom transfer carbonylation AB - [reaction: see text] Five- to seven-membered lactones were prepared from omega hydroxyalkyl iodides and CO by atom transfer carbonylation without the need for transition metal catalysts. The reaction proceeds via a hybrid radical/ionic mechanism in which the intramolecular alcoholysis of an omega-hydroxyacyl iodide, arising from atom transfer carbonylation, leads to the lactone. PMID- 10814331 TI - Fluoroacrylate-bound fluorous-phase soluble hydrogenation catalysts AB - [reaction: see text] Fluoroacrylate polymer-bound hydrogenation catalysts are described. N-Acryloxysuccinimide-containing fluoroacrylate polymers were converted into phosphine ligands and subsequently into analogues of Wilkinson's catalyst by amidation of N-acryloxysuccinimide active ester residues and Rh exchange. The resulting catalysts have excellent activity and can be reused following fluorous biphasic liquid/liquid separation and extraction. PMID- 10814332 TI - Reduction of azides to amines mediated by tin bis(1,2-benzenedithiolate) AB - [reaction: see text] A procedure for the conversion of azides to amines, which uses NaBH4 and catalytic amounts of tin(IV) 1,2-benzenedithiolate, is disclosed. Primary, secondary, tertiary, aromatic, and heteroaromatic azides are reduced in excellent yields under very mild conditions. PMID- 10814333 TI - Diastereoselectivity in Mn(III)-promoted 4-exo-trig cyclization of enamides to beta-lactams AB - [reaction: see text] The effect of chiral substituents on the enamide nitrogen atom upon the diastereoselection of the Mn(III)-mediated 4-exo-trig cyclization to beta-lactams was studied. A significant level of diastereoselectivity was achieved when an amino acid ester moiety was included into the enamidic skeleton. The structure of the major diastereoisomer was suggested by semiempirical calculations. PMID- 10814334 TI - Oxymercuration of homoallylic alcohol derived hemiacetals: diastereoselective synthesis of protected 1,3-diols. AB - [reaction: see text] Protected 1,3-diol synthons may be synthesized efficiently from homoallyic alcohols and simple aldehydes by oxymercuration of the derived hemiacetals. The reactions are diastereoselective and proceed without the use of solvent. Both Hg(OAc)2 and HgClOAc are effective in the reaction, and the latter produces isolable organomercurial chlorides directly. PMID- 10814335 TI - Synthesis of the N-((1E)-alkenyl)-(2Z,4Z)-heptadienamide side chain of salicylihalamide A and apicularens A and B. AB - [reaction: see text] The unstable N-((1E)-alkenyl)-(2Z,4Z)-heptadienamide side chain of salicylihalamide A (1) and apicularens A and B (3 and 4) has been prepared in one pot by the addition of (1Z,3Z)-hexadienylcuprate, prepared in situ from EtLi, CuBr.SMe2, and acetylene, to a (1 E)-alkenyl isocyanate. PMID- 10814336 TI - Concerted catalytic reactions for conversion of ketones or enol acetates to chiral acetates AB - [reaction: see text] Enol acetates or ketones asymmetrically transformed to chiral acetates in high yields with high optical purities through multistep reactions catalyzed by a lipase and a ruthenium complex. 2,6-Dimethylheptan-4-ol was chosen as a suitable hydrogen donor, and 4-chlorophenyl acetate was used as an acyl donor for the conversion of ketones. PMID- 10814337 TI - NaH-mediated one-pot cyclocondensation of 6-nitroquinoline with aromatic hydrazones to form AB - [reaction: see text] 6-Nitroquinoline undergoes direct cyclocondensation with aromatic hydrazones in the presence of sodium hydride in DMF at -10 degrees C, giving the corresponding 3-aryl-1(3)H-pyrazolo[3,4-f]quinolines and/or 3 aryl[1.2.4]triazino[6,5-f]quinolines in low to moderate yield. The mode of cyclocondensation is considerably dependent on the electronic nature of ring substituent of hydrazones, an electron-donating substituent favoring the formation of the latter heterocycles. PMID- 10814338 TI - Catalytic Asymmetric Synthesis of Diarylacetates and 4,4-Diarylbutanoates. A Formal Asymmetric Synthesis of (+)-Sertraline. PMID- 10814339 TI - A New Cryptand: Synthesis and Complexation with Paraquat. PMID- 10814340 TI - Fritsch-Buttenberg-Wiechell rearrangement in the aliphatic series AB - [reaction: see text] Substituted vinylidene zinc carbenoids undergo a Fritsch Buttenberg-Wiechell rearrangement. The migratory aptitude of the two groups R/R1 was studied by 13C labeling experiments and depends on the degree of substitution in R and R1 groups and on the presence of an oxygen atom in the allylic position. PMID- 10814341 TI - Unusual reactions of 5,5-dimethyl-2-(indenyl-2)-3-pyrazolidinone with acetylenedicarboxylates AB - [reaction: see text] Reaction of 5,5-dimethyl-3-pyrazolidinone (1) with 2 indanone (2) gave 5,5-dimethyl-2-(1H-indenyl-2)-3-pyrazolidinone (3) instead of the expected azomethine imine 4. Although reaction of 2-substituted 3 pyrazolidinones with acetylenedicarboxylates usually gives ring expansion products, such as 1,2-diazepines, treatment of 3 with dialkyl acetylenedicarboxylates (5, R = Me; 6 R = Et) resulted in the formation of rel (7aR,12aS)-6,7-bis(alkoxycarbonyl)-3,4-dihydro-4,4-dimethyl-7aH-indano[1,2 b]pyrrolo[1,2-a]pyrimidin-2-ones (7, R = Me; 9, R = Et) as major products and 3,4 bis(alkoxycarbonyl)-7,7-dimethyl-2-(indenyl-2)-6,7-dihydro-2H,6H-1,2-diazepin-5 ones (8, R = Me: 10, R = Et) as minor products. PMID- 10814342 TI - New spirans containing a 1,5-benzodithiepine system, derived from methylbenzenes. Conformational transmission AB - [structure: see text] A synthesis of three new spirans, derived from methylbenzenes, containing the 1,5-benzodithiepine system is reported. X-ray structure proved the identity of model monospiran 9. In the solid and liquid state, the conformation of the seven-membered ring is chair, and the five membered ring has the envelope conformation. The effect of conformational transmission in spirans 9 and 10 was observed. The synthesis of trispirans from hexamethylbenzene using the proposed scheme is also possible. PMID- 10814343 TI - Novel one-step synthesis of thiazolo[3,2-b]1,2,4-triazoles. AB - [reaction: see text] Reactions of chalcones 3a-f with bis(1H-1,2,4-triazolyl) sulfoxide 4 formed the thiazolo[3,2-b]1,2,4-triazoles 5a-f, which resemble closely some previously prepared COX-2 inhibitors. The structure of 5a was confirmed by X-ray analysis. PMID- 10814344 TI - Reactivity of quinoline- and isoquinoline-based heteroaromatic substrates in palladium(0)-catalyzed benzylic nucleophilic substitution AB - [reaction: see text] Quinolylmethyl, 1-(isoquinolyl)ethyl, and 1-(quinolyl)ethyl acetates reacted with dimethylmalonate anion in the presence of a Pd(0) catalyst to give products of nucleophilic substitution and/or byproducts, depending upon the substitution pattern. The observed side reactions were reduction in the case of primary acetates and elimination or elimination/Michael-type addition sequence for secondary substrates. PMID- 10814345 TI - Relative reactivities of tethered functional groups in the interior of a zeolite AB - [reaction: see text] The reactions of singlet oxygen with substrates containing both sulfide and olefinic linkages have been examined. The dramatic increase in sulfoxide formation in the interior of the zeolite in comparison to solution is attributed to zeolite stabilization of the persulfoxide intermediate and concomitant inhibition of physical quenching of singlet oxygen. PMID- 10814346 TI - Ruthenium-catalyzed cyclic carbonylation of allenyl alcohols. Selective synthesis of gamma- and delta-lactones AB - [reaction: see text] Ruthenium complex-catalyzed carbonylation of allenyl alcohols quantitatively gave cyclic carbonyl compounds, gamma- and delta lactones, in which the hydroxy group of allenyl alcohols participated in the cyclization. A wide variety of allenyl alcohols, such as mono-, di-, and trisubstituted alcohols, can be used in this reaction to produce 3- and 4 substituted gamma-lactones. Similarly, the cyclic carbonylation of 3,4-pentadien 1-ol 10a and 2-methyl-4,5-hexadien-2-ol 11a gave delta-lactones, 5,6-dihydro-3 methyl-2H-pyran-2-one 10b, and 5,6-dihydro-6,6-dimethyl-3-methyl-2H-pyran. 2-one 11b, respectively, in a quantitative yield. PMID- 10814347 TI - The sulfamate functional group as a new anchor for solid-phase organic synthesis AB - [reaction: see text] Sulfamate derivatives were loaded on trityl chloride resin, and two variants of cleavage were developed for this sulfamate anchor: an acid treatment to easily restore the free sulfamate and a nucleophilic treatment to generate the corresponding phenol. In addition to loading/cleavage assays and stability experiments, a model sequence of reactions was performed with the new sulfamate anchor to show its applicability in further combinatorial solid-phase synthesis of libraries of biologically relevant sulfamate derivatives. PMID- 10814348 TI - Macrocyclization and molecular interlocking via Mitsunobu alkylation: highlighting the role of C-H...O interactions in templating AB - [reaction: see text] A series of diimide-based macrocycles have been prepared using Mitsunobu-mediated alkylation as the macrocyclization step. These macrocycles could not be incorporated into [2]catenanes using previously established building blocks and coupling methodology. However, when one of the macrocycle syntheses was conducted in the presence of a dinaphtho crown ether, catenane formation was achieved. This result is discussed in terms of the ability of the components to establish intermolecular C-H...O hydrogen-bonding contacts. PMID- 10814349 TI - Electrosynthesis of alpha-arylated beta-substituted cyclopropylphosphonates. Synthesis of a phosphonic analogue of minalcipran AB - [reaction: see text] The synthesis of alpha-arylated beta-substituted cyclopropylphosphonates was efficiently achieved by electroreduction of diethyl alpha,alpha-dichlorobenzylphosphonate in the presence of Michael acceptors in a one-compartment cell equipped with a magnesium sacrificial anode. PMID- 10814350 TI - Facile ring transformation from gluconolactone to cyclitol derivative via spiro sugar ortho ester. AB - [reaction: see text] A short step preparation of cyclitol derivative 8 which is a versatile synthon for the synthesis of valiolamine and its related compounds is described. Key steps in this preparation are a novel enol ether formation from spiro sugar ortho esters with AlMe3 and an intramolecular Aldol condensation of alkyl enol ethers catalyzed by ZnCl2 in THF-H2O. With these reactions, gluconolactone derivative 1 was efficiently converted into 8 in short steps. PMID- 10814351 TI - A three-component coupling strategy for tetrahydrofuran synthesis: application of the diisopropyl tartrate modified (E)-gamma-(dimethylphenylsilyl)allylboronate as an alpha,gamma-allyl dianion equivalent. AB - [reaction: see text] A highly convergent three-component coupling strategy for the stereocontrolled synthesis of 2,3,5-trisubstituted tetrahydrofurans is described. After allylboration of the first aldehyde with 1, the chiral, nonracemic allylsilanes 2 are coupled with a second aldehyde or ketone with Lewis acid catalysis to give tetrahydrofurans 3 or 4 with excellent selectivity. The 2,5-stereochemistry is controlled by operating under nonchelate (e.g., 3) or chelate (e.g., 4) conditions. PMID- 10814352 TI - Asymmetric synthesis of (-)-indolizidines 167B and 209D based on stereocontrolled allylation of a chiral tricyclic N-acyl-N,O-acetal. AB - [reaction: see text] The tricyclic N-acyl-N,O-acetal incorporating (S)-2-(1 aminoethyl)phenol as a chiral auxiliary underwent TiCl4-mediated allylation to give the chiral (5S)-allylpyrrolidinone with retention of configuration in high yield and diastereoselectivity. On the bases of this methodology, the asymmetric syntheses of the dendrobatid alkaloids (-)-indolizidines 167B and 209D were achieved. PMID- 10814353 TI - Palladium-catalyzed cross-coupling of terminal alkynes with 4-trifloyloxazole: studies toward the construction of the C26-C31 subunit of phorboxazole A. AB - [reaction: see text] A strategy has been developed that successfully takes advantage of transition-metal-catalyzed coupling reactions for the synthesis of highly functionalized oxazoles. Trifloyloxazoles have been used as coupling partners with alkyne-derived vinylmetallic intermediates in Stille- and Negishi type couplings to assemble the corresponding oxazoles in good isolated yield. The results obtained provide a close analogy and thus good precedent to employ this strategy in the synthesis of the oxazole subunits of phorboxazole A. PMID- 10814354 TI - A new synthesis of p-hydroxy phenylglycine and some analogues from p-benzoquinone AB - [reaction: see text] A new route to p-hydroxy phenylglycine and N-substituted analogues has been developed starting from p-benzoquinone. 1,2-Addition of methyl lithioacetate to p-benzoquinone and subsequent quenching of the oxygen anion with methyl chloroformate, followed by an elimination-addition reaction with an appropriate amine, resulted in the desired amino acid derivatives. A diastereoselectivity of 60% was achieved using 8-phenylmenthyl acetate as the chiral auxiliary. PMID- 10814355 TI - A soluble polymer-supported triflating reagent: a high-throughput synthetic approach to aryl and enol triflates. AB - [reaction: see text] The high-yielding synthesis and application of the first example of a polymer-supported reagent for the preparation of trifluoromethanesulfonates (triflates) is described. This new reagent efficiently triflates aryl alcohols and lithium enolates in high yield (>90%). A simple precipitation and filtration to remove the excess reagent and byproduct facilitate purification of the triflate products. The PEG-supported approach is highly efficient, as the PEG-supported byproduct can be quantitatively recovered and recycled into reagent 1. PMID- 10814356 TI - CuCl-mediated intramolecular oxidative coupling of aryl- and alkenyltrimethylstannane functions AB - [reaction: see text] The syntheses of bis-trimethylstannanes 8, 10, 12-14, 16, 18, 21, and 22 are described. Treatment of these substances with approximately 5 equiv of CuCl in DMF at rt for 30-60 min effects, in each case, oxidative coupling between the two sp2 carbon centers bearing the Me3Sn function to produce good-to-excellent yields of tricyclic products 23-31, respectively. PMID- 10814358 TI - Facile O-deallylation of allyl ethers via S(N)2' reaction with tert-butyllithium AB - [reaction: see text] Allylic ethers are converted to the corresponding alcohol or phenol in virtually quantitative yield at temperatures below ambient simply by stirring a hydrocarbon solution of the ether with 1 molar equiv of tert butyllithium. The reaction, which produces 4,4-dimethyl-1-pentene as a coproduct, most likely involves an S(N)2' attack of the organolithium on the allyl ether. PMID- 10814357 TI - A convenient synthesis of 2-tetrahydrofuranyl ethers. AB - [reaction: see text] A wide spectrum of alcohols and phenols are readily transformed to the corresponding 2-tetrahydrofuranyl ethers in good to excellent yields using CrCl2 and CCl4 in THF under nearly neutral conditions at room temperature. PMID- 10814359 TI - Macrophyllols A and B, two unusual novel sesquiterpene and monoterpene dimers from the bark of Inula macrophylla. AB - [structure: see text] Two novel sesquiterpene and monoterpene dimers, macrophyllols A (1) and B (2), were isolated from the bark of Inula macrophylla. Their structures were determined on the basis of spectral evidence (especially HREIMS and 2D NMR) as well as chemical transformation. The structure of macrophyllol A (1) was confirmed by X-ray analysis. The possible biosynthetic pathways of macrophyllols A (1) and B (2) are discussed. PMID- 10814360 TI - Reductive aromatization of quinols: synthesis of the C-arylglycoside nucleus of the papulacandins and chaetiacandin. AB - [reaction: see text] Nucleophilic 1,2-addition of lithiated glycal 9b to functionalized quinone 7 provided, after reductive aromatization, C-arylglycoside 11b. Treatment with mCPBA afforded the tricyclic papulacandin framework. Alternatively, hydroboration gave the chaetiacandin nucleus. PMID- 10814361 TI - Enantioselective synthesis of syn- and anti-1,3-diols via allyltitanation of unprotected beta-hydroxyaldehydes AB - [reaction: see text] syn or anti-1,3-Diols units were synthesized with excellent diastereomeric excess from unprotected chiral beta-hydroxyaldehydes by using an enantioselective allyltitanation. PMID- 10814362 TI - Absolute configuration of scyphostatin AB - [reaction: see text] The absolute configuration of the side chain of scyphostatin (1) has been established. The chemical degradation of 1 gave 4 and 9, which correspond to the C7'-C12' and C13'-C16' fragments of the natural products, respectively. The spectroscopic data and [alpha]D values of both compounds were compared to those of authentic samples. The results show that the absolute configuration of 1 is 8'R,10'S,14'R. PMID- 10814364 TI - Photolabile dendrimers using o-nitrobenzyl ether linkages AB - [reaction: see text] Benzyl aryl ether dendrimers containing photosensitive, veratryl-based o-nitrobenzyl AB linkages (bold bonds) were prepared to the third generation and shown to undergo site-specific degradation when irradiated with ultraviolet light. PMID- 10814363 TI - A marine diterpene with a novel tetracyclic framework from the West Indian gorgonian octocoral Pseudopterogorgia elisabethae. AB - [structure: see text] Colombiasin A (1) was isolated from an extract of the West Indian gorgonian octocoral Pseudopterogorgia elisabethae that showed strong inhibitorial activity against Mycobacterium tuberculosis H37Rv. Structure elucidation by interpretation of 2D-NMR spectroscopic data, IR, UV, and accurate mass measurements (HREI-MS) revealed that colombiasin A belongs to a previously undescribed class of C20 rearranged diterpenes possessing an intricate tetracyclic framework. PMID- 10814365 TI - Absolute configuration of novel bioactive flavonoids from Tephrosia purpurea. AB - [structure: see text] Three novel flavonoids, (+)-tephrorins A (1) and B (2) and (+)-tephrosone (3), were isolated from Tephrosia purpurea. Their structures were elucidated by NMR spectral analysis, and their absolute configurations were determined by Mosher ester methodology. Compounds 1 and 2 are flavanones containing an unusual tetrahydrofuran moiety. Compounds 1-3 were evaluated for their potential cancer chemopreventive properties using a cell-based quinone reductase induction assay. PMID- 10814366 TI - A novel synthesis of 2-aryl-2H-indazoles via a palladium-catalyzed intramolecular amination reaction. AB - [reaction: see text] A variety of 2-aryl-2H-indazoles were synthesized by the palladium-catalyzed intramolecular amination of the corresponding N-aryl-N(o bromobenzyl)hydrazines. Of several sets of reaction conditions surveyed, the combination of Pd(OAc)2/dppf/tBuONa gave the best results. This method applies to a wide scope of substrates containing electron-donating and electron-withdrawing substituents. PMID- 10814367 TI - Acetalization of alkenes catalyzed by Pd(OAc)2/NPMoV supported on activated carbon under a dioxygen atmosphere AB - [reaction: see text] The acetalization of terminal alkenes such as ethyl acrylate and acrylonitrile with alcohols under O2 was efficiently achieved by Pd(OAc)2 supported on activated carbon combined with molybdovanadophosphate (NPMoV). For example, ethyl acrylate was subjected to acetalization with EtOH acidified by CH3SO3H under O2 (1 atm) in the presence of [8 wt%Pd(OAc)2/C] and NPMoV to form ethyl 3,3-diethoxypropionate in quantitative yield. PMID- 10814368 TI - A simple and efficient enantioselective synthesis of 2-alkylidene-3-alkyl-1,4 benzodioxanes by palladium-catalyzed annulation of benzene-1,2-diol and propargylic carbonates AB - [reaction: see text] Benzene-1,2-diol reacts with various propargylic carbonates in the presence of a palladium catalyst and a chiral atropoisomeric diphosphine to give 2-alkylidene-3-alkyl-1,4-benzodioxanes in good yields and 56-97% enantiomeric excess. PMID- 10814369 TI - Asymmetric aldol reactions on a soluble polymeric support AB - [reaction: see text] The combinatorial synthesis of small, nonpeptidic compounds is of increasing interest in current medicinal chemistry. To meet this demand, efficient entries, preferentially on polymeric supports, to pharmacologically interesting classes of compounds such as polyketides are necessary. Therefore, we have developed a synthetic protocol allowing for the asymmetric synthesis of diketides on the soluble support MeOPEG-5000. The strategy employed mainly allows for repeated aldolizations, thus providing access to functionalized polyketides of varying degree of oligomerization. PMID- 10814370 TI - Parallel kinetic resolution of racemic aldehydes by use of asymmetric Horner Wadsworth-Emmons reactions AB - [reaction: see text] A racemic aldehyde can undergo parallel kinetic resolution (PKR) by simultaneous reaction with two different chiral phosphonates, differing either in the structure of the chiral auxiliary or in the structure of the phosphoryl group (i.e., one (E)- and one (Z)-selective reagent). This strategy allows conversion of a racemic aldehyde to two different, synthetically useful chiral products with essentially doubled material throughput and similar or improved selectivities as compared to conventional kinetic resolution. PMID- 10814371 TI - A novel, general method for the synthesis of nitrile oxides: dehydration of O silylated hydroxamic acids AB - [reaction: see text] O-Silylated hydroxamic acids serve as stable, readily accessible, crystalline precursors to nitrile oxides when treated with trifluoromethanesulfonic anhydride and triethylamine. Under these mild conditions in the presence of olefins O-silylated hydroxamic acids afford isoxazoline cycloadducts. This procedure represents a novel, general method for the one-step generation of nitrile oxides, which complements existing protocols. PMID- 10814372 TI - Synthesis of eight-membered ring compounds using enyne metathesis AB - [reaction: see text] A novel procedure for synthesizing eight-membered ring compounds was developed using ruthenium-catalyzed enyne metathesis. When a CH2Cl2 solution of enyne connected with catechol, o-amino phenol, or o-phenylenediamine was stirred in the presence of benzylidene ruthenium carbene complex (10 mol %) at room temperature overnight, an eight-membered ring compound was obtained in high yield. In a similar manner, monocyclic 1,4-diaza- or 1-oxa-4-azacyclooctene derivative was obtained in high yield. PMID- 10814373 TI - Diastereoselective allylation of aldehydes with an enantiopure 2-sulfinylallyl halide under environmentally friendly Barbier-type conditions AB - [reaction: see text] A simple, efficient, and diastereoselective zinc-promoted allylation of aldehydes with enantiopure (Ss)-3-chloro-2-(p-tolylsulfinyl)-1 propene [(Ss)-1] under aqueous Barbier conditions is described. The observed diastereoselectivity can be explained via an acyclic antiperiplanar transition state model. PMID- 10814374 TI - Allene cross-metathesis: synthesis of 1,3-disubstituted allenes AB - [reaction: see text] Grubbs catalyst, Cl2(Cy3P)2Ru=CHPh, was found to catalyze the cross-metathesis of monosubstituted allenes to 1,3-disubstituted allenes in varying yields. PMID- 10814375 TI - Synthetic studies of the formation of oxazoles and isoxazoles from N-acetoacetyl derivatives: scope and limitations AB - [reaction: see text] The preparation of two types of heterocycles, oxazoles and isoxazoles, were achieved in good yields in a rapid and simple way by using N acetoacetyl derivatives. Steric and electronic effects caused by the nature of the substituents at C1, C2, and C3 were studied. The best results were obtained with a chiral oxazolidinone moiety on C1 derived from (1R,2S)-(-)-norephedrine. PMID- 10814376 TI - Conformational specificity in photoinduced intramolecular 1,7-hydrogen abstraction of homonaphthoquinones with a spiro-linked dibenzocycloheptene ring AB - [reaction: see text] Irradiation of homonaphthoquinones with spiro-linked dibenzocycloheptene rings brings about the Norrish type II reaction to give polycyclic alcohols via a stereospecific 1,7-hydrogen abstraction of the less stable twist-boat conformer. PMID- 10814377 TI - A novel and highly efficient synthesis of gem-difluorocyclopropanes AB - [reaction: see text] A new and highly versatile source of difluorocarbene is reported. Trimethylsilyl fluorosulfonyldifluoroacetate (TFDA) undergoes decomposition in the presence of catalytic fluoride to form difluorocarbene under conditions that allow its addition to relatively electron deficient alkenes in high yield. For example, unprecedented CF2: addition to n-butyl acrylate proceeded in 73% yield. PMID- 10814378 TI - Highly stereospecific, palladium-catalyzed cross-coupling of alkenylsilanols AB - [reaction: see text] Alkenylsilanols bearing methyl ((E)-1 and (Z)-1) or isopropyl ((E)-2 and (Z)-2)) substituents are converted to disubstituted alkenes by a palladium(0)-catalyzed cross-coupling reaction with aryl or vinyl iodides in the presence of tetrabutylammonium fluoride or hydroxide. Yields and stereoselectivities are generally high, and the reaction is compatible with a wide range of functional groups. PMID- 10814379 TI - Ligand Tuning in Asymmetric Catalysis: Mono- and Bis-Phospholanes for a Prototypical Pd-Catalyzed Asymmetric Allylation Reaction. PMID- 10814380 TI - A facile route to acyclic substituted alpha,beta-unsaturated aldehydes: the allene Claisen rearrangement AB - [reaction: see text] Sigmatropic rearrangement of allenyl ethers furnishes alpha,beta-unsaturated aldehydes in good yield. PMID- 10814381 TI - Stereochemistry of the cyclization-rearrangement of (+)-copalyl diphosphate to ( )-abietadiene catalyzed by recombinant abietadiene synthase from Abies grandis. AB - [reaction: see text] Syntheses and enzymatic cyclizations of 8alpha-hydroxy-17 nor copalyl diphosphate (8a), (15R)-[15-2H1] 8b, and (15R,17E)-[15-3H1,17-2H1] copalyl diphosphate ([2H,3H] 2) catalyzed by recombinant abietadiene synthase (rAS) gave 17-nor manoyl oxide (9a), (16E)-[16-2H1] 9b, and (15S,16R)-[16-2H1,16 3H1] abietadiene ([2H1,3H1] 4), respectively. These and other results indicate that conversion of CPP (2) to abietadiene (4) occurs by anti S(N)' cyclization to a sandaracopimar-15-en-8-yl carbocation intermediate (13+, 13beta-methyl) followed by hydrogen transfer and methyl migration suprafacially on the si face of the vinyl group. PMID- 10814382 TI - A catalytic oxidative esterification of aldehydes using V2O5-H2O2 AB - [reaction: see text] Aldehydes, in the presence of methanol, undergo oxidative transformation to the corresponding esters upon treatment with catalytic amounts of vanadium pentoxide in combination with oxidant hydrogen peroxide. Mild reaction conditions, shorter reaction times, high efficiencies, cost effectiveness, and facile isolation of the desired products make the present methodology a practical alternative. PMID- 10814383 TI - An efficient synthesis of thioisomunchnones derived from uracils and uridine: novel type of mesoionic nucleosides AB - [reaction: see text] Synthetic approaches to a variety of thioisomunchnones derived from uracils and uridine are described, as well as their properties and cycloaddition tendencies. The anhydro-3-hydrocy-2-phenyl-6-(2',3',5'-tri-O benzoyl-beta-D-ribofuranosyl)thiazolo[3,2-c]pyrimidine-5(6H)-on-4-ium hydroxide represents a novel class of mesoionic nucleosides. PMID- 10814384 TI - First asymmetric synthesis of a 6-alkoxy-5,6-dihydro-1,3-oxazine: a promising enantioselective route to beta-amido aldehydes AB - [reaction: see text] The 1,3-oxazine route to enantiopure beta-amido aldehydes was investigated. Heterocycloaddition of the N-acyl imine 1 with (R)-O vinylpantolactone provided the stable dihydroxazine 4c. High diastereocontrols were observed when using Yb(fod)3-catalyzed or SnCl4-mediated conditions, thus leading after quantitative hydrolysis to (R)-N-benzoyl-3-phenylpropanal with >98% ee. PMID- 10814385 TI - Visual sensing of saccharides promoted by resorcinol condensation products AB - [structure: see text] Heating aqueous DMSO solutions of five saccharides in the presence of 1-3 reveals that each receptor promotes solution colors characteristic for each sugar. New compound 3 allows the direct correlation of sugar concentration with visible region absorbance and/or fluorescence intensities. PMID- 10814386 TI - Rotaxanes via Michael addition AB - [reaction: see text] The application of a template effect to the classical Michael addition of heteronucleophiles to various Michael acceptor systems leads in good yields to a new trapping synthesis of [2]rotaxanes with conjugated functional groups in their axles. PMID- 10814387 TI - Synthesis of the C1-C13 fragment of leucascandrolide A. AB - [reaction: see text] The synthesis of the C1-C13 fragment 3 of leucascandrolide A has been completed utilizing a stereoselective and regioselective reductive cleavage of a highly functionalized spiroketal to incorporate the cis-2,6 disubstituted tetrahydropyan. The spiroketal was constructed by addition of a lithiated pyrone 5 to aldehyde 6. PMID- 10814388 TI - Structural diversity based on cyclopropane scaffolds AB - [structure: see text] A practical and efficient route for the stereoselective conversion of easily accessible homoallylic alcohols to diastereomerically pure cis- and trans-disubstituted and 1,2,3-trisubstituted cyclopropanes has been developed. The diversity of structures that can be prepared and the simplicity of the overall sequence make this route ideal for extension to solid-phase synthesis techniques and ultimately combinatorial library generation. PMID- 10814389 TI - A chiral sensor for arginine and lysine AB - [structure: see text] We provide access to a new class of C1- or C2-symmetrical host molecules 1 and 2 based on a spirobisindane skeleton. Whereas 1 is selective for short, rigid diamines, 2 prefers longer alpha,omega-dications. Of all the amino acid methyl esters, only those of lysine and arginine with the correct distance between their cationic groups form strong 1:1 complexes in DMSO with 2. NMR titrations reveal high association constants as well as discrimination between the enantiomers of lysine and arginine. PMID- 10814390 TI - Nitrogen-capped pi-prismands: syntheses and conformational analyses AB - [structure: see text] The one-pot syntheses of the new diazabicyclophanes 5-7 are described. The benzene rings incorporated in the bridging chains exhibit rotational motion which is examined by means of variable-temperature NMR experiments and semiempirical calculations. X-ray analysis and NMR studies indicate that the metal ion in the endohedral silver(I) complex of 7 fluctuates between the two nitrogen atoms. PMID- 10814392 TI - New C=C bond formation via nonstoichiometric titanium(IV) halide mediated vicinal difunctionalization of alpha,beta-unsaturated acyclic ketones AB - [reaction: see text] Highly stereoselective vicinal difuctionalization of alpha,beta-unsaturated ketones for the synthesis of multifunctionalized trisubstituted alkenes is described. The new reaction employs titanium(IV) halides (0.5 equiv) as promoters and inexpensive commercial chemicals as starting materials. The reaction can be performed at room temperature in a convenient vial without the protection of inert gases. Good to excellent yields and high Z/E stereoselectivity have been realized in most cases presented (16 examples). PMID- 10814391 TI - Characterization of spiroiminodihydantoin as a product of one-electron oxidation of 8-Oxo-7,8-dihydroguanosine. AB - [reaction: see text] Further oxidation of the common DNA lesion 8-oxo-7,8 dihydroguanosine by one-electron oxidants such as IrCl6(2-), Fe(CN)6(3-), or SO4 * leads to two major products, depending upon reaction conditions. In nucleosides at pH 7, 22 degrees C, the principal product is shown herein to be a spiroiminodihydantoin nucleoside, as a diastereomeric mixture, that can be characterized by NMR, ESI-MS/MS, and independent synthesis. PMID- 10814393 TI - Formal synthesis of (+/-)-peduncularine: use of the AB - [reaction: see text] The formal synthesis of the alkaloid (+/-)-peduncularine (1) was accomplished through the use of a [3 + 2] allylic silane annulation. The annulation of cyclohexadienyl silane 6 with chlorosulfonyl isocyanate followed by an in situ reduction provided bicyclic lactam 7. Conversion of this intermediate to 2 in three steps completed the formal synthesis of (+/-)-peduncularine. PMID- 10814394 TI - Synthesis of 2-oxazolidinones by direct palladium-catalyzed oxidative carbonylation of 2-amino-1-alkanols AB - [reaction: see text] 2-Oxazolidinones 2 are obtained in excellent yields (up to 100%) and with unprecedented catalytic efficiencies (up to 2000 mol of product/mol of catalyst used) by direct PdI2/KI-catalyzed oxidative carbonylation of the readily available 2-amino-1-alkanols 1. Reactions are carried out in MeOH as the solvent at 100 degrees C using a 1/6/5 CO/O2/air mixture (60 atm total pressure at 25 degrees C). PMID- 10814395 TI - C2-symmetric planar chiral ferrocene diamides by (-)-sparteine-mediated directed ortho-lithiation. Synthesis and catalytic activity AB - [reaction: see text] A variety of highly enantioenriched singly and doubly (4) functionalized derivatives of 1,1'-N,N,N',N'-tetraisopropylferrocenedicarboxamide 1 have been synthesized by (-)-sparteine (2)-mediated directed ortho-metalation and Pd-catalyzed cross coupling reactions. The synthetic applications of these chiral ligands in asymmetric alkylation of benzaldehyde and Pd(0)-catalyzed allylic substitution have been demonstrated. PMID- 10814396 TI - An efficient synthesis of pyrimidines from beta-amino alcohols. AB - [reaction: see text] Pyrimidinones 3 were chemoselectively reduced by using metal catalyzed hydrogenation and stereoselectively substituted by various nucleophiles. Starting from beta-amino alcohols 1, the overall process allows efficient access to substituted pyrimidines 4 and 6. PMID- 10814397 TI - Synthesis of (-)-dysiherbaine. AB - [reaction: see text] The first synthesis of (-)-dysiherbaine has been accomplished using intramolecular SN2 substitutions of a carbamate anion on an epoxide and an alkoxide on a secondary mesylate to efficiently construct the bicyclic skeleton stereospecifically from xylose. A general sequence has been developed to introduce an allyl group and convert it to the alanine side chain that should be useful for the construction of dysiherbaine analogues. PMID- 10814398 TI - A novel and efficient synthesis of 3-fluorooxindoles from indoles mediated by Selectfluor. AB - [reaction: see text] Treatment of several 3-substituted indoles, including derivatives of tryptophan and serotonin, with commercially available Selectfluor in acetonitrile/water furnished 3-substituted 3-fluorooxindoles in good to high yields. Since 3-fluorooxindoles obtained are sterically similar to both oxindoles and 3-hydroxyoxindoles, they should be useful as probes for investigating the enzymatic mechanism of indole biosynthesis and metabolism. PMID- 10814399 TI - Biomimetic total syntheses of (-)-TAN1251A, (+)-TAN1251B, (+)-TAN1251C, and (+) TAN1251D. AB - [reaction: see text] The muscarinic antagonists (-)-TAN1251A (1), (+)-TAN1251B (2), (+)-TAN1251C (3), and (+)-TAN1251D (4) have been synthesized biomimetically by enamine formation from an amino aldehyde to give TAN1251C ketal 18. Oxidation and reduction lead to TAN1251A (1), which has been hydroxylated to give TAN1251B (2). Stereospecific reduction of TAN1251C ketal 18 leads to TAN1251D (4). PMID- 10814400 TI - Treatment of N,N-dibenzylamino alcohols with sulfonyl chloride leads to rearranged beta-chloro amines, precursors to beta-amino acids, and not to tetrahydroisoquinolines. AB - [reaction: see text] Very recently, the unexpected formation of 3-substituted 1,2,3,4-tetrahydroisoquinolines starting from N,N-dibenzyl-protected beta-amino alcohols was reported. The authors claimed that treatment with tosyl chlorides induced intramolecular Friedel-Crafts alkylation. Reexamination of the reactions in our laboratory clearly proved rearranged chloro amines instead of the initially assumed tetrahydoisoquinoline structures. The chloro amines investigated can be employed as highly useful intermediates for an EPC synthesis of beta-amino nitriles and beta-amino acids. PMID- 10814401 TI - Formation of organomagnesium compounds via EtMgBr-mediated radical cyclization of allyl beta-iodoacetals AB - [reaction: see text] Treatment of allyl beta-iodoacetals with ethylmagnesium bromide in THF provided tetrahydrofuran derivatives in good yields. On the other hand, the reaction in DME provided tetrahydrofuranylmethylmagnesium compounds in good yields. PMID- 10814402 TI - Benzannulation from alkynes and allyl tosylates via a pi-allylpalladium intermediate AB - [reaction: see text] Allyl tosylate is a good allyl source for a novel palladium catalyzed benzannulation that affords polysubstituted benzenes from 1 mol of an allyl compound and 2 mol of internal alkynes. Using triphenyl phosphite as a ligand, the reaction with terminal alkynes gave trisubstituted benzenes regioselectively. PMID- 10814403 TI - Selective hydrogenation of benzophenones to benzhydrols. Asymmetric synthesis of unsymmetrical diarylmethanols AB - [reaction: see text] trans-RuCl2[P(C6H4-4-CH3)3]2(NH2CH2CH2NH2) acts as a highly effective precatalyst for the hydrogenation of a variety of benzophenone derivatives to benzhydrols that proceeds smoothly at 8 atm and 23-35 degrees C in 2-propanol containing t-C4H9OK with a substrate/catalyst ratio of 2000-20000. Use of a BINAP/chiral diamine Ru complex effects asymmetric hydrogenation of various ortho-substituted benzophenones and benzoylferrocene to chiral diarylmethanols with consistently high ee. PMID- 10814404 TI - Tandem oxy-Cope/transannular ene reaction of 1,2-divinylcyclohexanols AB - [reaction: see text] The syntheses via tandem oxy-Cope/transannular ene reaction of 1,2-divinylcyclohexanols to bi- and tricyclic skeletons are described. This strategy generates a rapid method for the preparation of advanced polycyclic intermediates with high diastereoselectivity. PMID- 10814406 TI - Formation of homoallyl stannanes via palladium-catalyzed stannylative cyclization of enynes AB - [reaction: see text] Treatment of 1,6-enynes with tri(n-butyl)tin hydride in the presence of a catalytic amount of palladium acetate leads to the formation of exo methylenecyclopentanes bearing a tri(n-butyl)stannylmethyl moiety. PMID- 10814405 TI - Fullerene-sensitized AB - [reaction: see text] Fullerene catalyzes the cycloaddition of dimethyliminodiacetate to maleimides under photolysis to form 2,5 dimethoxycarbonylpyrrolidine derivatives. PMID- 10814407 TI - An efficient synthesis of optically active physostigmine from tryptophan via alkylative cyclization. AB - [reaction: see text] A new and efficient synthetic route to physostigmine is described. Corey-Kim reagent reacted with tryptamine or tryptophan carbamates to give 3a-(methylthiomethyl)hexahydropyrrolo[2,3-b]indole skeletons. Formal total synthesis of racemic and chiral physostigmine was accomplished in excellent overall yields, in short steps. PMID- 10814408 TI - A modular approach to marine macrolide construction. 3. Enantioselective synthesis of the C1-C28 sector of spongistatin 1 (altohyrtin A). AB - [structure: see text] A completely stereocontrolled approach to assembly of the major C1-C28 subunit of spongistatin 1 (altohyrtin A) is described. Key steps included the control of two asymmetric aldols by means of Fujita-Nagao (chiral N acyl-1,3-thiazolidine-2-thione auxiliary) and Mukaiyama (BF3 x OEt2-promoted enolsilane coupling) protocols in complex settings. PMID- 10814409 TI - A novel coumarin-type derivatizing reagent of alcohols: application in the CD exciton chirality method for microscale structural determination AB - [reaction: see text] Imidazolide 1 has been prepared from 7-diethylaminocoumarin 3-carboxylic acid (2) and 1,1'-carbonyldiimidazole. This novel fluorescent derivatization reagent can be used in CD exciton chirality method for microscale structural determinations of hydroxyl-containing compounds. It features a red shifted chromophore and offers the advantages of fluorescence and high sensitivity. PMID- 10814410 TI - The chemistry of acylals. 3. Cyanohydrin esters from acylals with cyanide reagents AB - [reaction: see text] When treated with KCN in DMSO at room temperature, acylals from aliphatic aldehydes gave the corresponding cyanohydrin esters in good to excellent yields. Acylals from aromatic aldehydes were less reactive and gave several byproducts in addition to fair yields of cyanohydrin under the same conditions. Trimethylsilyl cyanide mixed with titanium(IV) chloride afforded cyanohydrin esters in good to excellent yields from both aliphatic and aromatic aldehydes. PMID- 10814411 TI - Segregation of separate steps in chromium-catalyzed reactions for convenience and mechanistic analysis AB - [structure: see text] We have developed an apparatus for carrying out redox coupled chromium-manganese or chromium-zinc reactions on solid-supported substrates which allows recycling of the stoichiometric reductant. In addition to providing a greatly simplified procedure for these reactions, we observe striking differences in the recyclability of Mn and Zn. A preliminary extension of the methodology to the catalytic coupling of allylchromium reagents with solid supported aldehydes is also reported. PMID- 10814412 TI - Mechanistic studies on selectivity in the B(C6F5)3-catalyzed allylstannation of aldehydes: is hypercoordination at boron responsible? AB - [structure: see text] The selective, B(C6F5)3-catalyzed allylstannation of aldehydes with proximal donor groups is shown to proceed via borane abstraction of the allyl group from the tin reagent and activation of the substrate by "Bu3Sn+". This is supported by a number of 19F NMR experiments. The selectivity of the reaction is not attributable to hypercoordinate boron as proposed by the discoverers of this highly selective reaction but likely involves chelation at tin. PMID- 10814413 TI - A 9-hydroxyiridoid isolated from Junellia seriphioides (Verbenaceae). AB - [structure: see text] An unusual iridoid glucoside, namely 9-hydroxy-8 epihastatoside (1), was isolated from Junellia seriphioides (Verbenaceae), together with the known compounds auroside (2), pulchelloside I (3), and 8 epihastatoside (4) as well as verbascoside. PMID- 10814414 TI - The use of diazabicyclo AB - [reaction: see text] In a study directed toward the use of the chloroacetyl protecting group in carbohydrate synthesis, the sterically hindered tertiary amine diazabicyclo[2.2.2]octane (DABCO) was found to give complete and selective cleavage of the chloroacetyl group in the presence of other ester functions such as benzoyl and acetyl groups at primary and/or secondary positions. PMID- 10814415 TI - A short total synthesis of (+)-furanomycin. AB - [reaction: see text] Furanomycin is a Streptomyces metabolite that substitutes for isoleucine in protein translation. We report a concise and modular synthesis starting from the Garner aldehyde and proceeding in seven steps to furanomycin. The key steps include a stereoselective acetylide addition and the Ag+-mediated cyclization of an alpha-allenic alcohol to construct the trans-2,5-dihydrofuran. The efficiency (12% overall yield) and flexibility of the route will provide ample quantities of furanomycin and analogues for protein engineering. PMID- 10814416 TI - Pairwise use of complexity-generating reactions in diversity-oriented organic synthesis. AB - [reaction: see text] Two pairs of complexity-generating reactions with an essential product-substrate relationship along a synthetic pathway are demonstrated. This pathway illustrates a key element in a planning algorithm for diversity-oriented synthesis. This element facilitates the efficient synthesis of structurally complex compounds, and it can be integrated with ones that provide structurally diverse compounds. PMID- 10814418 TI - Stereoselective bis-functionalizations of arene chromium tricarbonyl complexes via brook rearrangements AB - [reaction: see text] An efficient method has been developed for the stereoselective bis-functionalization of arene chromium tricarbonyl complexes. Initial nucleophilic addition of organolithium reagents to a carbonyl moiety is followed by a 1,4-carbon to oxygen silyl migration (Brook rearrangement) and alkylation of the resultant aryl anion. PMID- 10814417 TI - A method for the asymmetric hydrosilylation of N-aryl imines. AB - [reaction: see text] The asymmetric reduction of N-aryl imines to yield chiral amines with enantiomeric excesses above 90% was achieved. Ethylenebis(eta5 tetrahydroindenyl)titanium difluoride ((EBTHI)TiF2, 1) was employed as the precatalyst with polymethylhydrosiloxane (PMHS) as the stoichiometric reducing agent. A variety of N-aryl imines derived from nonaromatic ketones were reduced with high ee. PMID- 10814419 TI - In-Line Proximity Effects in Extended 7-Azanorbornanes. 1. A New Concept for Modifying Effector Group Separation Based on the Control of N-Invertomer Geometry. AB - Control of N-substituent geometry in fused 7-azanorbornane systems is based on the dominance of one proximate bridge (sentinel X) over the other (sentinel Y) relative to the N-bridge; the N-inversion equilibrium can effectively be displaced in favor of a single invertomer. This study has used a combination of synthesis, crystallography, and molecular modeling to establish stereostructures. PMID- 10814420 TI - In-Line Proximity Effects in Extended 7-Azanorbornanes. 2. A Major Reduction of N Inversion Barriers in Symmetrically Flanked Systems. AB - Through-space compression effects from a pair of flanking (sentinel) groups X are shown to cause a flattening of the bridging nitrogen in symmetrically fused 7 azanorbornanes which is evaluated by a substantial reduction of the nitrogen inversion barrier as measured by low-temperature (1)H NMR spectroscopy. This effect is attributed to destabilization of the ground state of the amine. PMID- 10814421 TI - Diastereoselective Baylis-Hillman Reactions: The Design and Synthesis of a Novel Camphor-Based Chiral Auxiliary. AB - Reaction of chiral acryloylhydrazide 5 derived from novel auxiliary 4 with aldehydes in the presence of DABCO affords practical levels (up to 98% de) of beta-hydroxy-alpha-methylene carbonyl derivatives, and high optical purity of both diastereomers of 6a-d and 7a-d can be obtained by changing the solvent. PMID- 10814422 TI - Synthesis and Diels-Alder Reactions of a New Kind of Chiral Dienophiles: Cyclic Vinyl-p-tolylsulfilimines. AB - A new kind of chiral dienophiles, cyclic vinyl-p-tolylsulfilimines (2a and 2b), were obtained from the corresponding (Z)-sulfinylacrylonitriles with HBF(4) and methanol. The asymmetric Diels-Alder reaction of optically pure 2a with cyclopentadiene under mild thermal or catalyzed conditions afforded only the endo 4a adduct with complete endo and pi-facial selectivities. The ability of the sulfilimine moiety to enhance the dienophilic reactivity of the double bond is similar to that of the sulfinyl group. PMID- 10814423 TI - Chemo- and Regioselective Dimerization of Terminal Alkynes Promoted by Methylaluminoxane. AB - Methylalumoxane (MAO) was found to be an active catalytic precursor for the chemo and regioselective dimerization of a wide range of aryl- and alkyl-substituted terminal alkynes yielding the corresponding geminal dimers A. For an olefin functionalized terminal alkyne (RC&tbd1;CH, R = MeC=CH(2)), the geminal dimer undergoes an intermolecular [4 + 2] cycloaddition forming compound B. PMID- 10814424 TI - A New Type of Amido-Substituted p-tert-Butylcalix AB - A convenient method for the synthesis of intramolecularly bridged calix[6]arenes by bis(chloroacetyl)amide was reported. A 1,3-singly bridged derivative and an asymmertical 1,3-4,5 doubly bridged derivative were obtained, respectively. PMID- 10814425 TI - Diastereoselective Formation of Methylene-Bridged Glycoluril Dimers. AB - The acid-catalyzed formation of methylene-bridged glycoluril dimers yields the C(2)(v)()-diastereomer selectively. Product resubmission experiments establish that the selectivity is the result of thermodynamic control. A modified synthetic route is presented that allows for the preparation of unsymmetrically substituted dimers. We present the X-ray crystal structures of both diastereomers. This class of compounds is useful for studies of self-assembly in aqueous solution. PMID- 10814426 TI - Toward Daisy Chain Polymers: "Wittig Exchange" of Stoppers in AB - Two ammonium ion/crown ether-based [2]rotaxane monomers-each incorporating (i) a dumbbell-shaped component, possessing an exchangeable benzylic triphenylphosphonium stopper, and (ii) a ring component, bearing an aldehyde function-undergo a sequence of Wittig reactions in which the surrogate triphenylphosphonium stopper is exchanged for a ring component either (i) in the same rotaxane molecule to give cyclic daisy chains by an intramolecular, chain terminating reaction or (ii) in another rotaxane molecule to give acyclic daisy chains by an intermolecular chain-propagating reaction. PMID- 10814427 TI - Synthesis, Structure, and Complexing Property of p-tert-Butylcalix AB - p-tert-Butylcalix[4]arene 1,3-digallate, which contains a nonbonded close contact between galloyl groups, was synthesized and its structure was determined by dynamic (1)H NMR and X-ray crystallography. The electronic spectra showed that a new absorption band of the complexes appeared at a longer wavelength region upon adding Ag(+) ion. This spectral shift was explained in terms of the interaction between the facing galloyl groups of the lower rim of the calixarene framework. PMID- 10814428 TI - Solvent-Dependent Lithium Bridging in Allenyl-Propargyllithium Reagents. AB - The solution structures of two allenyl-propargyllithium reagents (6-Li and 7-Li) which give (13)C NMR chemical shifts intermediate between those expected for the allenyl and propargyl isomers have been studied by the Saunders isotope perturbation technique. Variable equilibrium isotope shifts were detected, showing that these reagents adopt structures ranging from equilibrating localized allenyl- and propargyllithium reagents (1A right harpoon over left harpoon 1B) to equilibrating unsymmetrically bridged structures (11) and symmetrically bridged structures (2), depending mostly on the details of solvation. PMID- 10814429 TI - Unusual AB - All-carbon 1,3-dipoles derived from allenic esters/ketones, by catalytic interaction with triphenylphosphine, undergo an unusual [8 + 2] annelation with tropone, leading to 8-oxa-9-(ethoxycarbonyl/acylalkylidene)bicyclo[5.3.0]deca 1,3,5-trienes. Dipoles derived from allenic ketones as well as an alpha-methyl substituted allenic ester display high reactivity and selectivity. PMID- 10814430 TI - 1-Cyanoimidazole as a Mild and Efficient Electrophilic Cyanating Agent. AB - A mild and high-yielding cyanating reaction of amine, sulfur, and carbanion nucleophiles is reported here using 1-cyanoimidazole as an electrophilic cyanating agent. PMID- 10814431 TI - Efficient Electrochemical Deprotection of Carboxylic and Amino Acids from Their 2 (Hydroxymethyl)-1,3-dithiane (Dim) Esters. AB - Carboxylic acids and amino acids are electrochemically deprotected from their 2 (hydroxymethyl)-1,3-dithiane (Dim) esters. PMID- 10814432 TI - Unusual C-6 Lithiation of 2-Chloropyridine-Mediated by BuLi-Me(2)N(CH(2))(2)OLi. New Access to 6-Functional-2-chloropyridines and Chloro-bis-heterocycles. AB - The reaction of 2-chloropyridine with alkylithium generally results in nucleophilic addition leading to the loss of chlorine atom while exclusive directed ortho metalation is obtained using LDA. Herein it is shown that the BuLi Me(2)N(CH(2))(2)OLi (BuLi-LiDMAE) superbase promotes an unprecedented regioselective C-6 lithiation. The method was successfully applied to the preparation of potentially useful chlorinated pyridinic and bis-heterocyclic synthons. PMID- 10814433 TI - Cobalt-Induced C-N and C-C Bond Formation via Metal-Stabilized alpha-CF(3) Carbenium Ion. AB - An alpha-CF(3)-carbenium ion stabilized by a bimetallic [Co-Co] cluster was prepared and isolated in good yield, starting from the corresponding alcohol by action of HBF(4)/Et(2)O. C-N and C-C bonds with nitrogen and carbon nucleophiles could be easily formed. Subsequent decomplexation gave the free substituted beta CF(3) alkynes in good yields. PMID- 10814434 TI - Aberrant S(RN)1 Reaction of 4-Aminophenol with alpha,p-Dinitrocumene: EPR Observation of Intermediates. AB - The tert-butoxide-induced substitution of alpha,p-dinitrocumene by 4-aminophenol unexpectedly afforded the N-coupled product, 2-(4-hydroxyanilino)-2-(4 nitrophenyl)propane. EPR observations revealed arylaminyl radical intermediates as well as coupled anion radicals, hence the normal S(RN)1 process may compete with an alternative nonchain reaction pathway. PMID- 10814435 TI - Hyperconjugative Control by Remote Substituents of Diastereoselectivity in the Oxygenation of Hydrocarbons. AB - The oxidation of 2-substituted adamantanes (2) with TFDO (1) is reported. The data show a stereodifferentiation of the chemical environments induced by remote electron-withdrawing substituents which produces remarkable Z/E diastereoselectivity in the oxidation of the tertiary C(5)-H and C(7)-H bonds. The results show a bell-shaped correlation between the Z/E stereoselectivity and the substituent constant sigma(I), which is interpreted in terms of hyperconjugative stabilization of the diastereomeric transition states. PMID- 10814436 TI - Rate Constant for the Ring Opening of the 2,2-Difluorocyclopropylcarbinyl Radical. AB - The rate constant for the unimolecular ring opening of the 2,2 difluorocyclopropylcarbinyl radical was determined via its competitive bimolecular trapping by TEMPO. The value of this rate constant (3.4 x 10(11) s( )(1) at 99.3 degrees C) is about 500 times larger than that of the parent, unfluorinated radical and about 5 times smaller than that of the trans-2 phenylcyclopropylcarbinyl radical. PMID- 10814437 TI - Calix Crowns Derived from para-Bridged Calix AB - Two calix[4]arenes, bridged by aliphatic chains of five and eight carbon atoms spanning two opposite para positions, have been converted into their 1,3-crown ether derivatives by reaction with tetraethylene and pentaethylene glycol ditosylates. Of the two possible 1,3-crown derivatives, only the formation of those isomers is observed in which the phenolic units connected at the para positions are etherified. This has been established in one case by a single crystal X-ray analysis. Preliminary results for the extraction of selected alkali and alkaline-earth metals are reported. PMID- 10814438 TI - Dendrimers Based on Melamine. Divergent and Orthogonal, Convergent Syntheses of a G3 Dendrimer. AB - Both convergent and divergent strategies are used to synthesize 1, a dendrimer comprising triazines linked by diamines. The convergent approach is orthogonal; neither protecting groups nor functional group manipulations are required using the building blocks selected. PMID- 10814439 TI - A New Preparative Method for Allylic Indium(III) Reagents by Reductive Transmetalation of pi-Allylpalladium(II) with Indium(I) Salts. AB - A reductive transmetalation of the pi-allylpalladium(II) complexes, generated in situ from a catalytic amount of a palladium(0) complex and a variety of allylic substrates, with indium(I) salts proceeded smoothly in various solvents, providing a new route for allylindium(III) reagents. PMID- 10814440 TI - Two Useful Photolabile Surfaces for Solid-Phase Synthesis. AB - o-Nitrobenzyl-based photolabile surfaces 3 and 4 have been synthesized from 2 phenylcyclohexanone and aminopropylsiloxane-grafted controlled pore glass. The procedures are simple and inexpensive and generate materials whose minimally functionalized reactive chromophore should tolerate a range of subsequent chemistries. Time- and solvent-dependent analysis of photodegradation demonstrates a performance comparable to related photolabile systems. PMID- 10814442 TI - Synthesis of Bicyclic Cyclopropylamines by Intramolecular Cyclopropanation of N Allylamino Acid Dimethylamides. PMID- 10814441 TI - Complexation-Initiated Intramolecular Diels-Alder Reaction. AB - A complexation-initiated intramolecular Diels-Alder reaction was demonstrated for the first time. On adsorbing the alkyne-Co(2)(CO)(6) complexes having a diene and a dienophile on opposite ends of the alkyne on silica gel, the Diels-Alder reaction was accelerated and the equilibrium was shifted toward the adduct formation to a greater extent compared to that in solution. Seven-membered ring formation was apparently favorable in this system. PMID- 10814443 TI - beta-scission of the N-O bond in alkyl hydroxamate radicals: a fast radical trap. AB - [reaction--see text ] The rate of the beta-scission of the N-O bond in the alkyl hydroxamate radical is faster than 2 x 10(8) s(-)(1). This reaction may be useful as a radical trap. PMID- 10814444 TI - Reductive cleavage of N-O bonds using Samarium(II) iodide in a traceless release strategy for solid-phase synthesis AB - A strategy for making amides and ureas using a polymer-supported hydroxylamine resin as a traceless linker is described. The cleavage of the linker by samarium(II) iodide is reported for the first time. PMID- 10814445 TI - A cyclodextrin-based molecular shuttle containing energetically favored and disfavored portions in its dumbbell component AB - A molecular shuttle containing dodecamethylene units, 4, 4'-bipyridinium units, and alpha-cyclodextrin (alpha-CD) has been prepared. Shuttling behavior is solvent- and temperature-sensitive and could be controlled by double interactions: hydrophobic interaction between a CD ring and a station and a repulsive interaction between a CD ring and a linker. This is a novel method to control the mobility of a bead in a molecular shuttle. PMID- 10814446 TI - Laser flash photolysis of 2-diazo-1,3-diphenyl-1,3-propanedione: An unusual long lived triplet as a reaction intermediate AB - Laser flash photolysis of 2-diazo-1,3-diphenyl-1,3-propanedione (DBD) is presumed to involve a short-lived carbene, followed by Wolff rearrangement to a long-lived ketene. We have detected ketene ylides following photolysis of DBD in the presence of amines but not with pyridine. The triplet state of DBD lives several microseconds, an unusual observation for a diazo compound; however, the triplet is not a ketene precursor, which must result from excited singlet state fragmentation of DBD. PMID- 10814447 TI - Cross-linked enzyme aggregates: a simple and effective method for the immobilization of penicillin acylase. AB - [reaction--see text] Penicillin G acylase (penicillin amidohydrolase, E.C. 3.5.1.11) was immobilized in a simple and effective way by physical aggregation of the enzyme, using a precipitant, followed by chemical cross-linking to form insoluble cross-linked enzyme aggregates (CLEAs). These had the same activity in the synthesis of ampicillin as cross-linked crystals of the same enzyme, but the accompanying hydrolysis of the side-chain donor was much less. Penicillin G acylase CLEAs also catalyzed the synthesis of ampicillin in a broad range of organic solvents. PMID- 10814448 TI - Using polarization effects to alter chemical reactivity: A simple host which enhances amine nucleophilicity AB - The rational design of a bis(phosphine oxide) host which is capable of binding a benzylic amine is presented. The ability of this host to increase the rate of addition of 4-fluorobenzylamine to N-phenylmaleimide is rationalized in terms of the enhancement of the nucleophilicity of the benzylic amine. PMID- 10814449 TI - Site specificity in the photooxidation of some trisubstituted alkenes in thionin supported zeolite Na-Y. On the role of the alkali metal cation AB - The less substituted side of some geminal dimethyl trisubstituted alkenes becomes significantly more reactive if photooxygenation takes place in thionin-supported zeolite Na-Y. These results are mainly attributed to a synergistic interaction between the alkali metal cation, the alkene, and the oxygen in the transition state of perepoxide formation. PMID- 10814450 TI - Mass transport limitations reduce the effective stereospecificity in enzyme catalyzed kinetic resolution. AB - [reaction-see text] The kinetic resolution of seudenol catalyzed by Candida antarctica lipase B in hexane was investigated. Large differences in reaction rate and stereospecificity were observed when different enzyme preparations were used. These differences were ascribed to mass transport limitations which reduced both reaction rate and stereospecificity. Lyophilized enzyme preparations were more apt to give this problem than immobilized preparations. Further, low substrate concentrations enhanced the effect. Thus, high alcohol concentrations and enzyme immobilization can be recommended. PMID- 10814452 TI - Benzhydryldimethylsilyl allylic silanes: syntheses and applications to AB - A new silyl group, the benzhydryldimethylsilyl group, has been developed that is easily synthesized and that undergoes facile oxidation. The [3 + 2] annulation reactions of allylic silanes with this silyl group provide a variety of highly substituted five-membered carbocycles and heterocycles with high stereoselectivities. The silyl groups of these cyclic compounds have been oxidized to hydroxyl groups to demonstrate the general synthetic utility of the method. PMID- 10814451 TI - Synthesis of unsymmetrical diarylmethanes by cross-coupling between aryl triflates and tetrabutylammonium difluorotribenzylstannate AB - The benzylation of aryl triflates can be achieved by cross-coupling between aryl triflates and the new hypervalent tin reagent (n-Bu(4)N)(+)(Bn(3)SnF(2))(-). PMID- 10814453 TI - A novel pathway in the photooxygenation of cyclic allenes. AB - [equation--see text] The photooxidation of cyclic allenes gives rise to cyclic 1,2, 3-trione hydrates. The formation of these compounds points to a novel photooxidation mechanism involving both singlet and triplet oxygen. Upon placement of a methyl group on the allene, the mechanism shifts to predominantly an "ene" reaction. The corresponding cycloadditions with 4-methyl-1,3,4 triazoline-3, 5-dione (MTAD) with cyclic allenes involve 2 equiv of MTAD. The dipolar intermediates are trapped with H(2)O to give alpha-urazole-substituted 2 cycloalkenones. PMID- 10814454 TI - One-pot construction of medium- and large-sized ring substituted furans. Efficient conversion to dibenzofurans, coumestans, and 4-pyrones. AB - New efficient synthesis of medium- and large-sized ring substituted furans is achieved by 1,3-dicarbonyl compounds with vinyl sulfides in the presence of Ag(2)CO(3)/Celite (Fetizon's reagent) in a one-pot procedure. The synthesized furans can be further converted to biologically interesting compounds such as dibenzofurans, coumestans, benzofuroquinolinone, and 4-pyrone. PMID- 10814455 TI - Synthesis of the ezomycin nucleoside disaccharide. AB - [equation--see text] A protected ezomycin octosyl nucleoside was glycosylated at O-6' with a protected ezoaminuroic acid donor to afford, following several functional group modifications, the title compound 1 ( identical with 4-desamino 4-oxoezomycin A(2)). PMID- 10814456 TI - Enantioselective synthesis of the pyrroloquinoline core of the martinellines. AB - [equation--see text] The first enantioselective synthesis of the martinelline core (-)-3 is reported. The synthesis of (-)-3 from N-allyl-N-(benzyloxycarbonyl) 2-iodoaniline (12) proceeded in seven steps and 23% overall yield. In addition, the preparation of a carbocyclic model system is described. PMID- 10814457 TI - Synthesis of N-hydroxy peptides: chemical ligation of O-acyl hydroxamic acids. AB - [process--see text] A novel chemical ligation process is described that results in the construction of N-hydroxy peptides. PMID- 10814458 TI - Efficient palladium-catalyzed N-arylation of indoles. AB - [equation--see text] The N-arylation of indoles, including a variety of substituted ones, has been carried out using bulky, electron-rich phosphines as the supporting ligand in combination with Pd(2)(dba)(3). Using this catalyst system, the efficient coupling of indole and a variety of substituted indoles with aryl iodides, bromides, chlorides, and triflates can be achieved. PMID- 10814459 TI - Total synthesis of the cyathane diterpenoid (+/-)-sarcodonin G. AB - [process--see text] The total synthesis of (+/-)-sarcodonin G (3), a highly functionalized member of the cyathane family of diterpenoids, is described. PMID- 10814460 TI - Regio- and stereocontrolled metal-mediated carbonyl propargylation or allenylation of enantiomerically pure azetidine-2,3-diones: synthesis of highly functionalized 3-substituted 3-hydroxy-beta-lactams AB - Regio- and stereocontrolled metal-mediated Barbier-type reactions of azetidine 2,3-diones with differently substituted propargyl bromides offer an efficient asymmetric entry to densely functionalized 3-propargyl- (or allenyl-) substituted 3-hydroxy-beta-lactams. PMID- 10814461 TI - Toward catalytically active oligonucleotides: synthesis of a flavin nucleotide and its incorporation into DNA. AB - [reaction--see text] The synthesis of a vitamin B(2)-derived flavin-nucleotide is described. A combined H-phosphonate/phosphoramidite protocol was developed for the first incorporation of flavin coenzymes into a DNA stack. The coenzyme-DNA is predicted to have novel biosensing and catalytic properties. PMID- 10814462 TI - Palladium(0)-catalyzed cyclization reaction of polymer-supported aryl iodides with 1,2-allenyl carboxylic acids. A facile solid-phase synthesis of butenolides. AB - [reaction--see text] Aryl iodides anchored to Merrifield resin via the linkages between a carboxyl, hydroxyl, or hydroxymethyl group and the benzylic chloride moiety in the resin reacted efficiently with 1,2-allenic carboxylic acids under the catalysis of Pd(0) to afford polymer-supported butenolides. Polysubstituted butenolides can be easily cleaved from the resins by Lewis acid-catalyzed processes in good yields and purities. PMID- 10814464 TI - Imposing curvature on a polyarene by intramolecular palladium-catalyzed arylation reactions: A simple synthesis of Dibenzo AB - Palladium catalysis has been found to offer an effective solution-phase alternative to gas-phase flash vacuum pyrolysis as a method for converting the planar ring system of 7, 10-di(2-bromophenyl)fluoranthene (4) to that of the C(28)H(14) bowl-shaped fullerene fragment dibenzo[a,g]corannulene (5). PMID- 10814463 TI - High turnover number and rapid, room-temperature amination of chloroarenes using saturated carbene ligands. AB - [equation--see text] A catalytic system for the mild amination of aryl chlorides is described. This system consists of a Pd(0) precursor and a dihydroimidazoline carbene ligand, which is generated in situ from its protonated tetrafluoroborate salt (2). Using this catalyst, aryl and heteroaryl chlorides react with secondary amines and anilines within hours at room temperature. Turnover numbers as high as 5000 are obtained at elevated temperatures for reaction of morpholine with an unactivated aryl chloride. PMID- 10814465 TI - Synthesis of bis- and oligo-gem-difluorocyclopropanes using the olefin metathesis reaction AB - Synthesis of six types of novel bis- and oligo-gem-difluorocyclopropanes has been accomplished through the olefin metathesis reaction protocol. Two types of ruthenium-based olefin metathesis catalysts were tested: the ruthenium catalyst coordinated with 1,3-dimesityl-4,5-dihydroimidazol-2-ylidene and tricyclohexylphosphine ligands gave better results than the ruthenium catalyst that coordinated with two tricyclohexylphosphine ligands. PMID- 10814466 TI - The effect of varying substituents on the equilibrium distribution and conformation of macrocyclic steroidal N-acyl hydrazones AB - The equilibrium distribution of deoxycholate-based macrocyclic N-acyl hydrazones is dependent upon the nature of the substituents at the 3- and 12-position. These substituents can also influence the preferred conformation of the macrocycles. The conversion of pure cyclic dimer and trimer into a mixture of both components, in the presence of trifluoroacetic acid, has been demonstrated and an X-ray crystal structure obtained of a macrocyclic N-acyl hydrazone dimer. PMID- 10814467 TI - Synthesis of the apoptosis inducing agent apoptolidin. Assembly of the C(16) C(28) fragment. AB - [reaction--see text] A stereoselective synthesis of the C(16)-C(28) fragment of the apoptosis inducing agent apoptolidin is described. Key steps include two propionate aldol reactions and a stereoselective Mukaiyama aldol addition of enolsilane 19 to beta-methoxy aldehyde 4. PMID- 10814468 TI - Stereocontrol in solid-phase radical reactions: radical addition to oxime ether anchored to polymer support AB - A high degree of stereocontrol in solid-phase radical reactions was achieved by using triethylborane and diethylzinc as a radical initiator at low reaction temperature. Alkyl radical addition to Oppolzer's camphorsultam derivatives of oxime ether anchored to polymer support proceeded smoothly to give the alpha amino acid derivatives with excellent diastereoselectivities. PMID- 10814469 TI - Novel deprotection of SEM ethers: A very mild and selective method using magnesium bromide AB - New lability and stability sequences have been established for multifunctional substrates containing SEM ether group(s) by a MgBr(2)/Et(2)O/MeNO(2) deprotection protocol. PMID- 10814470 TI - The 2-(N,N-Dimethylamino)phenylsulfinyl group as an efficient chiral auxiliary in intramolecular heck reactions AB - The synthesis, reactivity, and stereochemical behavior of differently substituted iodoalkenyl alpha,beta-unsaturated sulfoxides in intramolecular Heck reaction is described. Particularly, the 2-(N,N-dimethylamino)phenylsulfinyl group is demonstrated to be an effective chiral auxiliary for the intramolecular Heck reaction of 2-iodo-1,6-(or 1,7)dienes. A desulfinylation sequence removes the auxiliary and yields cyclic compounds of high enantiopurity. PMID- 10814471 TI - Preparation of alcohols from alkenes via the homologation of boronates with (Trimethylsilyl)diazomethane AB - Alkylcatecholboranes obtained from alkenes were converted to the corresponding alkylmethanols by reaction with (trimethylsilyl)diazomethane followed by oxidation and treatment with fluoride. PMID- 10814472 TI - An unusual samarium diiodide mediated reductive ring contraction of a tricyclic oxazine to a highly-functionalized cyclopentane and cyclobutane. AB - [reaction--see text] Samarium diiodide mediated reductive ring contraction of a substituted tricyclo[2.2.2]oxazine at 25 degrees C leads to a mixture of cyclopentane and cyclobutane rearrangement products with complete diastereoselectivity in each case. At -78 degrees C, the anticipated amidocyclohexanol reduction product is obtained exclusively, while the cyclopentane is the sole product at reflux in THF. PMID- 10814473 TI - An enantioconvergent route to (-)-shikimic acid via a palladium-mediated elimination reaction. AB - [reaction--see text] (-)-Shikimic acid, the key intermediate in the shikimate pathway in plants and microorganisms, has been synthesized in an enantioconvergent manner from both enantiomeric starting materials by employing a palladium-mediated elimination reaction as the key step. PMID- 10814474 TI - Solid-phase synthesis of acridine-peptide conjugates and their analysis by tandem mass spectrometry. AB - [reaction--see text] A novel and high-yielding synthesis of 9-anilinoacridine-4 carboxylic acid is reported. This acid has been used in the solid-phase synthesis of a small combinatorial library of acridine-peptide conjugates. Tandem mass spectrometry (ES-MS/MS) can be used for structure determination of these compounds at a sensitivity of approximately 10 pmol. This work makes possible the generation of acridine-peptide libraries for the discovery of structure-specific nucleic acid ligands via affinity chromatography selection with mass spectrometric detection. PMID- 10814475 TI - Use of pentamethyldisiloxane in the palladium-catalyzed cyclization/hydrosilylation of functionalized dienes. AB - [reaction--see text] Pentamethyldisiloxane reacts with a range of functionalized dienes in the presence of a catalytic 1:1 mixture of (N-N)Pd(Me)Cl [N-N = 1, 10 phenanthroline or (R)-(+)-4-isopropyl-2-(2-pyridinyl)-2-oxazoline] and NaBAr(4) [Ar = 3,5-C(6)H(3)(CF(3))(2)] to form the corresponding silylated carbocycles in good yield and with good stereoselectivity. Treatment of these silylated carbocycles with excess KF and peracetic acid at room temperature for 48 h formed the corresponding alcohols in excellent yield with retention of stereochemistry. PMID- 10814476 TI - Type 2 intramolecular nitroso Diels-Alder reaction. Synthesis and structure of bridgehead oxazinolactams. AB - [reaction--see text] The type 2 intramolecular Diels-Alder cycloaddition utilizing N-acylnitroso dienophiles provides an efficient entry into bridged oxazinolactams. In contrast to the bimolecular counterpart, the reaction is completely regioselective. Structural characterization of the cycloadducts allows for evaluation of the olefin distortion and the degree of pyramidalization of the bridgehead oxazinolactam. PMID- 10814477 TI - gamma-hydroxy unsaturated nitriles: chelation-controlled conjugate additions. AB - [reaction--see text] Chelation between gamma-hydroxy unsaturated nitriles and Grignard reagents promotes an otherwise difficult anionic conjugate addition reaction. The intermediate chelate is readily generated by deprotonation with t BuMgCl followed by the addition of a second Grignard reagent that triggers an intramolecular conjugate addition. Structurally diverse Grignard reagents add with equal efficiency, providing an intermediate anion that stereoselectively alkylates BnBr in an overall addition-alkylation reaction. PMID- 10814478 TI - Synthesis (and alternative proof of configuration) of the scyphostatin C(1') C(20') trienoyl fragment. AB - [reaction--see text] Each of four diastereomers of structure 2, corresponding to the lipophilic side chain of scyphostatin (1), were prepared. Careful analysis of their NMR spectral data and comparison with those of the natural product corroborates the recently reported (Org. Lett. 2000, 2, 505) stereochemical assignment. A strategy for the stereoselective synthesis of 2 has been achieved. PMID- 10814479 TI - A general method for acylation of indoles at the 3-position with acyl chlorides in the presence of dialkylaluminum chloride. AB - [reaction--see text] Indoles are selectively acylated at the 3-position in high yields on treatment with a wide variety of acyl chlorides in CH(2)Cl(2) in the presence of diethylaluminum chloride or dimethylaluminum chloride. The reaction proceeds under mild conditions and is applicable to indoles bearing various functional groups without NH protection. PMID- 10814480 TI - Phenyl migrations in dehydroaromatic compounds. A new mechanistic link between alternant and nonalternant hydrocarbons at high temperatures AB - Flash vacuum pyrolysis of benzo[b]biphenylene, an alternant polycyclic aromatic hydrocarbon (PAH), gives fluoranthene, a nonalternant PAH, as the major product at 1100 degrees C in the gas phase. The most reasonable mechanism to explain this isomerization involves equilibrating diradicals of 2-phenylnaphthalene that rearrange by the net migration of a phenyl group to give equilibrating diradicals of 1-phenylnaphthalene, one isomer of which then cyclizes to fluoranthene. PMID- 10814482 TI - A Formalism to Study the Propagation of the Four-Wave Mixing Signal in a Strongly Driven Two-Level System Immersed in a Thermal Reservoir. AB - We analyze two analytical solutions for the study of the propagation of the four wave mixing (FWM) signal in a strongly driven two-level system, when the stochastic effects of the solvent are explicitly considered. The first solution is valid only for a constant pump intensity, while the second solution considers pump propagation explicitly. In both cases, we were able to derive analytical expressions for the FWM nonlinear intensity signal in terms of the field amplitude and thermal noise parameters. Copyright 2000 Academic Press. PMID- 10814483 TI - Intensity Measurements and Collision-Broadening Coefficients for the Oxygen A Band Measured by Intracavity Laser Absorption Spectroscopy. AB - High-sensitivity, high-resolution intracavity laser absorption spectroscopy (ICLAS) has been used to measure line intensities, nitrogen-broadening coefficients, and self-broadening coefficients in the A band (b(1)Sigma(+)(g) <-- X(3)Sigma(-)(g)) of oxygen. Both linear cavity and ring cavity ICLAS configurations were used for these measurements, and the results were intercompared. The results were compared to values measured using long-path multiple-reflection cells by K. D. Ritter and T. D. Wilkinson [J. Mol. Spectrosc. 121, 1-19 (1987)] and L. Brown and C. Plymate, [J. Mol. Spectrosc. 199, 166-179 (2000)]. New results are included for weakly absorbing transitions, not observed in the earlier measurements, such as high rotational states (up to J = 39), hot band transitions (v' = 1 <-- v" = 1), and isotopically substituted species ((18)O(2) and (16)O(18)O). Isotopic variants ((16)O(2), (18)O(2), and (16)O(18)O) have similar broadening coefficients for corresponding rotational levels, but the self-broadening coefficients are larger in the hot band (v' = v" = 1) as compared with v' = v" = 0 transitions. An ECS-EP scaling analysis of the v' = v" = 0 self broadening data accurately represents the available data, with the exception of the N = 0 and N = 1 levels. Copyright 2000 Academic Press. PMID- 10814481 TI - The first total synthesis of a highly branched arabinofuranosyl hexasaccharide found at the nonreducing termini of mycobacterial arabinogalactan and lipoarabinomannan. AB - [reaction--see text] The first total synthesis of the arabinofuranosyl hexasaccharide present at the nonreducing termini of mycobacterial arabinogalactan and lipoarabinomannan is reported. The oligosaccharide was prepared as its methyl glycoside via a route that is both highly efficient and convergent. Addition of two beta-D-arabinofuranosyl residues simultaneously in high yield and with excellent stereocontrol was the key step of the synthesis. PMID- 10814484 TI - Vibrational Excitations of Methane in the Framework of a Local-Mode Anharmonic Model. AB - The stretching and bending vibrations of methane are described in a local-mode anharmonic symmetry adapted model. Both Hecht- and Fermi-like interactions are considered. We start by establishing the Hamiltonian of the system in terms of internal coordinates and momenta. The Hamiltonian is then rewritten in terms of local creation (annihilation) operators a(dagger)(i)(a(i)), which are later anharmonized by means of the deformation of the harmonic oscillator algebra to the U(2) algebra. This procedure corresponds to the exact Morse result for the dominant matrix elements involved in the harmonically coupled anharmonic oscillator model (HCAO) introduced by Child and Lawton. We have carried out a fit to 40 experimental levels with an rms deviation of 1.17 cm(-1). In addition, we present a description of the dipole transition intensities. Copyright 2000 Academic Press. PMID- 10814485 TI - Air- and N(2)-Broadening Parameters of Water Vapor: 604 to 2271 cm(-1). AB - High-resolution measurements of air- and N(2)-broadened widths and pressure induced frequency shifts of water vapor were obtained covering the spectral region between 604 and 2271 cm(-1). Over 1300 vibration-rotation transitions were measured including the (000)-(000), (010)-(010), (010)-(000), (020)-(010), and (100)-(010) vibrational bands of H(2)(16)O. Also included were measurements of H(2)(18)O and H(2)(17)O from normal water vapor samples and H(2)(18)O + N(2) observations with oxygen-18-enriched gas samples. Collision-narrowing effects were observed in a few lines involving high J and low K(a) transitions with the lowest measured linewidth coefficient equal to 0.0057(4) cm(-1)/atm (air broadening) for the completely overlapping transitions, 17 0 17 <-- 18 1 18 and 17 1 17 <-- 18 0 18, at 1235.204 cm(-1). The majority of the linewidth values were derived from the measurements using a Voigt line profile. A few lines with air- or N(2)-broadened half-width values of about 0.012 cm(-1) or less were found to exhibit collision-narrowing effects and were also analyzed with a profile proposed by Galatry. The results are compared to values given in previous studies. Copyright 2000 Academic Press. PMID- 10814486 TI - Characterization of Electronic Spectra of Rhodium Monohydride and Monodeuteride. AB - The electronic spectra of rhodium monohydride and monodeuteride, obtained using a molecular beam laser vaporization source, have been investigated from 400 to 500 nm, by laser excitation and dispersed fluorescence spectroscopies. The ground level of RhH is (3)Delta(3) with omega(e) approximately 2040 cm(-1) and omega(e)x(e) approximately 46 cm(-1). The corresponding RhD data are omega(e) approximately 1422 cm(-1) and omega(e)x(e) approximately 18 cm(-1). The estimated ground state bond length is r(0) = 0.159 nm. Excited states with Omega = 2, 3, and 4 have been observed. Copyright 2000 Academic Press. PMID- 10814487 TI - Line Intensities of (12)C(16)O(2) in the 1.2-1.4 um Spectral Region. AB - The 7000-8500 cm(-1) spectral region of (12)C(16)O(2) has been investigated using the high-resolution FT spectrometer of LPPM in Orsay. The two strongest bands in this region are the 10031 <-- 00001 and 10032 <-- 00001 bands centered at 8294 and 8192 cm(-1). Line intensities in these two bands and in the 40013 <-- 00001 and 40014 <-- 00001 bands have been measured. Using the method of effective operators, these line intensities have been included in a new fit of effective dipole-moment parameters to all available experimental data in the same spectral region of (12)C(16)O(2). The corresponding calculated line intensities of the 10031 <-- 00001 and 10032 <-- 00001 bands are compared with the experimental ones. Copyright 2000 Academic Press. PMID- 10814488 TI - A Dense Grid of Reference Iodine Lines for Optical Frequency Calibration in the Range 595-655 nm. AB - A dense grid of reference lines in the hyperfine structure of the B-X transitions of molecular iodine ((127)I(2)) is presented. Frequencies of 481 "t"-hyperfine components in 15 vibrational bands covering the wavelength range 595-655 nm were determined with an absolute accuracy of 1.0 MHz (1varsigma uncertainty). Spectra were obtained by Doppler-free saturation spectroscopy using a cw ring-dye laser of 1-MHz bandwidth. Center-of-gravity frequencies of rovibronic lines, determined by correcting for the calculated hyperfine shift, are included separately in a least-squares parametrization for each band. From the obtained molecular constants a prediction is made for all t-hyperfine components of the 15 bands covering 595-655 nm. Hence a dense grid of reference lines of 1 MHz (1varsigma) absolute accuracy is created, with a line in each interval of 1 cm(-1) in the entire range. This spectral atlas of about 3400 lines may provide a useful calibration tool in many laboratories. Copyright 2000 Academic Press. PMID- 10814489 TI - Fourier Transform Emission Spectroscopy of the Low-Lying Electronic States of NbN. AB - The high-resolution spectrum of NbN has been investigated in emission in the 3000 15 000 cm(-1) region using a Fourier transform spectrometer. The bands were excited in a microwave discharge through a mixture of NbCl(5) vapor, approximately 5 mTorr of N(2), and 3 Torr of He. Numerous bands observed in the near-infrared region have been classified into the following transitions: f(1)Phi c(1)Gamma, e(1)Pi-a(1)Delta, C(3)Pi(0+)-A(3)Sigma(-)(1), C(3)Pi(0-)-A(3)Sigma( )(1), C(3)Pi(1)-a(1)Delta, C(3)Pi(1)-A(3)Sigma(-)(0), d(1)Sigma(+)-A(3)Sigma( )(0), and d(1)Sigma(+)-b(1)Sigma(+). These observations are consistent with the energy level diagram provided by laser excitation and emission spectroscopy [Y. Azuma, G. Huang, M. P. J. Lyne, A. J. Merer, and V. I. Srdanov, J. Chem. Phys. 100, 4138-4155 (1993)]. The missing d(1)Sigma(+) state has been observed for the first time and its spectroscopic parameters are consistent with the theoretical predictions of S. R. Langhoff and W. Bauschlicher, Jr. [J. Mol. Spectrosc. 143, 169-179 (1990)]. Rotational analysis of a number of bands has been obtained and improved spectroscopic parameters have been extracted for the low-lying electronic states. The observation of several vibrational bands with v = 1 has enabled us to determine the vibrational intervals and equilibrium bond lengths for the A(3)Sigma(-)(0), a(1)Delta, b(1)Sigma(+), d(1)Sigma(+), and C(3)Pi(1) states. Copyright 2000 Academic Press. PMID- 10814490 TI - High-Resolution Analysis of the nu(4) Absorption Band of CH(2)(79)BrF. AB - The gas-phase infrared spectrum of the nu(4) fundamental band of CH(2)(79)BrF was recorded in the 1010-1116 cm(-1) wavenumber region using a TDL spectrometer. In this first high-resolution investigation of the synthesized (79)Br isotopic form, more than 10 200 transitions of this a/b-hybrid band centered at 1068.5385 cm(-1) were assigned and, using the Watson's A-reduced Hamiltonian in the I(r) representation, a reliable set of molecular constants for the excited state v(4) = 1 was determined. From ground state combination differences having rotational quantum numbers J and K(a) up to 97 and 21, respectively, improved and extended ground state rotational and centrifugal distortion constants were calculated as well. Comparison between the observed and calculated band intensities in appropriate regions of the spectrum gave an estimate of the transition dipole moment ratio along the a and b axes as ||Deltau(a)/Deltau(b) || = 2.0 +/- 0.2, in agreement with the predicted theoretical value of 1.99. Copyright 2000 Academic Press. PMID- 10814491 TI - High-Resolution Infrared Spectrum of the Ring-Puckering Band, nu(10), of Diborane. AB - The spectrum of the nu(10) band of diborane, arising from the ring-puckering vibration, has been obtained with a spectral resolution of 0.0015 cm(-1) in the region 275-400 cm(-1). The spectrum of a sample enriched in (10)B was recorded as well as one with naturally abundant boron, i.e., 64% (11)B(2)H(6), 32% (10)B(11)BH(6), and 4% (10)B(2)H(6). This mode is the lowest vibrational level of the molecule and is unperturbed, allowing a complete assignment of not only the fundamental bands but also the 2nu(10)-nu(10) hot bands of all three boron isotopomers. The intensities of several hundred lines of the fundamental and hot bands of all isotopomers have been measured and vibrational transition moments have been obtained. Finally, it has been shown that the harmonic approximation does not apply for nu(10). Copyright 2000 Academic Press. PMID- 10814492 TI - The nu(12) Band of CH(3)SiD(3). AB - The lowest frequency degenerate fundamental band of CH(3)SiD(3) (v(12) = 1 <-- 0) centered around 418 cm(-1) was measured in order to investigate the vibration torsion-rotation interactions in a symmetric-top molecule with a single torsional degree of freedom. The spectrum was recorded at an instrumental resolution of 0.004 cm(-1) using a Bomem Fourier transform spectrometer. The temperature and pressure of the sample were 180 K and 2 Torr, respectively. Because of the Coriolis coupling between the torsional stack with one quantum of the silyl rock excited and the corresponding stack for the ground vibrational state, torsional splittings are measured that are substantially larger than expected simply from the observed increase in the barrier height. Due to the local nature of the Coriolis perturbation, the significantly enhanced torsional splittings are confined to a few (K, varsigma) rotational series; here varsigma = -1, 0, 1 labels the torsional sublevels. The current measurements of the nu(12) band and frequencies from previously reported studies in the ground vibrational state were fitted to within experimental uncertainty using an effective Hamiltonian which was used for the analyses of similar spectra in CH(3)SiD(3) and CH(3)CD(3). Spectroscopic parameters characterizing the states v(12) = 0 and 1 and their interactions were determined, including several Coriolis-coupling constants. Copyright 2000 Academic Press. PMID- 10814493 TI - Experimental and Ab Initio Studies of the HDO Absorption Spectrum in the 13 165 13 500 cm(-1) Spectral Region. AB - The HDO absorption spectrum was recorded in the 13 165-13 500 cm(-1) spectral region by intracavity laser absorption spectroscopy. The spectrum (615 lines), dominated by the 2nu(2) + 3nu(3) and nu(1) + 3nu(3) bands, was assigned and modeled leading to the derivation of 196 accurate energy levels of the (103) and (023) vibrational states. Finally, 150 of these levels were reproduced by an effective Hamiltonian involving two vibrational dark states interacting with the (023) and (103) bright states. The rms deviation achieved by variation of 28 parameters is 0.05 cm(-1), compared to an averaged experimental uncertainty of 0.007 cm(-1), indicating the limit of validity of the effective Hamiltonian approach for HDO at high-vibrational excitation. The predictions of previous ab initio calculations of the HDO spectrum (H. Partridge and D. Schwenke, J. Chem. Phys. 106, 4618-4639 (1997)) were extensively used in the assignment process. The particular spectral region under consideration was used to test and discuss the improvements of new ab initio calculations recently performed on the basis of the same potential energy surface but with an improved dipole-moment surface. The improvements concern both the energy levels and the line intensities. In particular, the strong hybrid character of the nu(1) + 3nu(3) band is very well accounted for by the new ab initio calculations. Copyright 2000 Academic Press. PMID- 10814494 TI - The Coriolis Interaction between the nu(9) and nu(7) Fundamental Bands of Methylene Fluoride. AB - The infrared spectrum of the nu(7) and nu(9) bands of methylene fluoride-d(2) (CD(2)F(2)) has been recorded with an unapodized resolution of 0.0024 cm(-1) in the frequency range of 940-1030 cm(-1) using the Fourier transform technique. A weak b-type Coriolis interaction term was found to couple these two vibrational states with band centers about 42 cm(-1) apart. By fitting a total of 1031 infrared transitions of both nu(7) and nu(9) with a standard deviation of 0.0011 cm(-1) using a Watson's A-reduced Hamiltonian in the I(r) representation with the inclusion of a b-type Coriolis resonance term, two sets of rovibrational constants for nu(7) = 1 and nu(9) = 1 states up to sextic order were derived. The nu(7) band is C type, while the nu(9) band is A type with band centers at 961.8958 +/- 0.0005 and 1003.7421 +/- 0.0001 cm(-1), respectively. Copyright 2000 Academic Press. PMID- 10814495 TI - Hyperfine Level Dependence of the Pressure Broadening of CH(3)I Rotational Transitions in the v(6) = 1 Vibrational State. AB - We studied the hyperfine components of the (J = 10-9, Kl = 9) rotational transition in the v(6) = 1 excited vibrational state of CH(3)I, using collinear infrared and mm-wave radiations. The Doppler-free double-resonance technique allowed an accurate determination of the collisional broadening parameters for all the hyperfine components. An evident dependence on the F quantum number was observed and this result is perfectly consistent with a theoretical model allowing calculations of collisional broadening and coupling for the hyperfine components. Copyright 2000 Academic Press. PMID- 10814496 TI - An IAM Calculation of the Torsional-Rotational Term Values for Selected Species of -OH and -OD Ethyl Alcohol. PMID- 10814497 TI - An IAM Calculation of the Torsional-Rotational Term Values for Three Isotopic Species of Ethyl Mercaptan. PMID- 10814498 TI - Pure Rotational Spectrum of Ethylenimine at 1.85 THz. PMID- 10814499 TI - First Observation of the BaI A' (2)Delta Electronic State: The Omega = 3/2 Component. PMID- 10814501 TI - Polyamines: mysterious modulators of cellular functions. AB - In recent years the functions of polyamines (putrescine, spermidine, and spermine) have been studied at the molecular level. Polyamines can modulate the functions of RNA, DNA, nucleotide triphosphates, proteins, and other acidic substances. A major part of the cellular functions of polyamines can be explained through a structural change of RNA which occurs at physiological concentrations of Mg(2+) and K(+) because most polyamines exist in a polyamine-RNA complex within cells. Polyamines were found to modulate protein synthesis at several different levels including stimulation of special kinds of protein synthesis, stimulation of the assembly of 30 S ribosomal subunits and stimulation of Ile tRNA formation. Effects of polyamines on ion channels have also been reported and are gradually being clarified at the molecular level. PMID- 10814502 TI - Neprilysin II: A putative novel metalloprotease and its isoforms in CNS and testis. AB - Metalloproteases of the M13 subfamily, comprising namely neprylisin (NEP) and endothelin-converting enzyme (ECE), are involved in the metabolism of various neuronal and hormonal peptides, and inhibitors thereof have already led to therapeutically useful agents. Using homology cloning, we have identified a new member of this family in rat tissues. It is a glycosylated, type II integral membrane protein of 774 amino acids, containing a zinc-binding consensus motif, highly homologous to NEP and, therefore, designated NEPII. We have characterized multiple splice variants of NEPII mRNA with distinct expression patterns in brain regions, pituitary and testis. In situ hybridization of testis, where levels of the NEPII gene transcript are the highest, reveals a localization within round spermatids. In brain, NEPII is expressed heterogeneously among several neuronal populations and according to a pattern grossly complementary to that of NEP. PMID- 10814503 TI - Peroxisome proliferator-activated receptor-gamma agonists increase vascular endothelial growth factor expression in human vascular smooth muscle cells. AB - Vascular endothelial growth factor (VEGF), expressed in a variety of mesenchymal cells including vascular smooth muscle cells (VSMC), is a potent mitogen for endothelial cells, and is used clinically applied for ischemic disease of peripheral vessels. To determine whether peroxisome proliferator-activated receptor gamma (PPARgamma) regulates VEGF production in VSMC, we examined VEGF secretion from VSMC treated with PPAR agonists. Troglitazone increased VEGF secretion in a time- and dose-dependent manner (261 +/- 35% with 25 mM of troglitazone for 24 h), and also increased levels of VEGF mRNA. VEGF secretion was also increased by other PPARgamma agonists, pioglitazone, LY171883, and 15d PGJ2 (224 +/- 17.1%, 247 +/- 36.8% and 171 +/- 7.8%, respectively), but not the PPARgamma agonists bezafibrate and Wy14643 (85.2 +/- 1.5%, 94.6 +/- 3.2, respectively). Our findings suggest that thiazolidinediones might be useful for the therapeutic angiogenesis for ischemic artery disease. PMID- 10814504 TI - Identification of human PDE7B, a cAMP-specific phosphodiesterase. AB - We isolated a human cAMP-specific phosphodiesterase (PDE7B) cDNA from human caudate nucleus. The human PDE7B was composed of 450 amino acid residues with a molecular mass of 51,835 Da. The deduced amino acid sequence of human PDE7B was 64.1% identical to that of human PDE7A (67.1% identity in the catalytic region). Northern blot analysis demonstrated that PDE7B transcripts were abundantly expressed in the putamen, caudate nucleus, and heart followed by skeletal muscle, pancreas, and occipital pole. Recombinant PDE7B expressed in transfected COS-7 cells had a low cAMP K(m) value of 0. 13 microM, which is similar to the K(m) value of recombinant human PDE7A expressed in transfected COS-7 cells. Interestingly, the relative V(max) value of recombinant PDE7B was half to one third of recombinant PDE7A. The PDE7B activity was inhibited by dipyridamole and SCH51866, with IC(50) values of 1.1 microM and 1.5 microM, respectively. Thus, the PDE7B exhibited unique tissue distribution in humans and kinetic profiles. Human PDE7B showed the lowest K(m) values compared to the other cAMP-hydrolyzing PDEs which have been reported to be expressed in the brain. Therefore, human PDE7B may be involved in the control of cAMP-mediated neural activity and cAMP metabolism in the brain. PMID- 10814505 TI - Integrin-associated protein and thrombospondin-1 as endothelial mechanosensitive death mediators. AB - Recently, it was reported that the offset of hemodynamic forces induces an unusual pattern of apoptosis in vascular endothelium (1). Although the apoptotic trigger covers all cells and is maintained for a longer time period, only few cells become apoptotic. So, in contrast to common apoptosis inducers, the lack of hemodynamic forces initiates only a low basal level of apoptosis, however steadily increases with time, this way preventing the complete vessel destruction upon an only transient offset of blood flow. The molecular means by which the mechanical stimulus and apoptosis are smoothly coupled have now been identified as an autocrine loop of thrombospondin-1 (TSP-1) and the alpha(v)beta(3) integrin/integrin-associated protein (IAP) complex as its receptor. Vascular EC (EC) secrete TSP-1 only in postconfluent static monolayers and not under flow. This also holds true for the IAP whereas the alpha(v)beta(3) integrin is present under static conditions, as well as under flow, assigning the IAP an essential and new switch function in the receptor complex. PMID- 10814506 TI - Phenotypic effects of CD3zeta targeting into glycosphingolipid-enriched membrane microdomains (GEMs) of T cells. AB - In the present study we tested whether the forced expression of the CD3zeta chain within detergent-resistant, glycosphingolipid-enriched membrane microdomains (GEMs) will result in a constitutively activated phenotype in human T cells. To this aim, a monomeric recombinant protein (LckSH4-CD3zeta), containing the intracellular part of human CD3zeta chain fused to N-terminal double-acylation motif (SH4 domain) of protein tyrosine kinase Lck, was expressed in Jurkat human T lymphoid cell line and its Lck-negative mutant, J. CaM1.6. The Lck SH4 domain indeed predominantly targeted the chimeric protein into GEMs. In transfectants derived from wild-type Jurkat cells, but not in those derived from the Lck deficient mutant, the LckSH4-CD3zeta protein was constitutively tyrosine phosphorylated. Tyrosine phosphorylation of a major Jurkat cell phosphoprotein (pp85) was diminished in the transfectants. However, the transfectants did not exhibit any features of constitutively activated T cells, and their responses to anti-CD3 treatment were very similar to the wild-type Jurkat cells. Thus, the constitutive expression of this form of CD3zeta chain in GEMs is not sufficient for eliciting an activated state in the Jurkat cells. PMID- 10814507 TI - Role of Raf-1 conserved region 2 in regulation of Ras-dependent Raf-1 activation. AB - Full activation of Raf-1 requires the interaction of its CRD with Ras. The serine/threonine-rich region, CR2, of Raf-1 was implicated in Raf-1 regulation, but the underlying mechanism was unclear. Here we show that CRD loses its Ras binding activity when expressed in connection with CR2, suggesting that CR2 masks CRD. This masking effect is abolished by substitution of Asp or Ala for Ser-259, a growth factor- and TPA-induced phosphorylation site in CR2. Treatment of COS-7 cells expressing Ha-Ras(Val-12) and Raf-1 with TPA enhances the Ha-Ras(Val-12) dependent Raf-1 kinase activity. In contrast, the Ha-Ras(Val-12)-dependent activities of the Raf-1(S259D) and Raf-1(S259A) mutants are comparable to that of wild-type Raf-1 stimulated by both Ha-Ras(Val-12) and TPA and cannot be further stimulated by TPA treatment. These results suggest that the in vivo phosphorylation of Ser-259 may comprise a crucial step for Ras-dependent Raf-1 activation by unmasking CRD and promoting its association with Ras. PMID- 10814508 TI - Novel mix-family homeobox genes in zebrafish and their differential regulation. AB - We report the isolation of two novel zebrafish mix-type homeobox genes, mtx1 and mtx2. The homeodomains of both Mtx1 and Mtx2 exhibited a 50% amino acid identity to other Mix-family protein homeodomains. mtx1 was expressed throughout the yolk syncytial layer (YSL), an extraembryonic structure in teleosts, from the late blastula to the mid-gastrula period. mtx2 was first expressed in the dorsal blastomeres soon after the mid-blastula transition, and slightly later in the entire blastoderm margin. After the late blastula period, mtx2 transcripts were detected in the YSL, and they were restricted to the dorsal YSL by the early gastrula period. The expression of mtx2 was dependent on Wnt signals but not on Nodal signals. mtx1 expression was not regulated by either Wnt or Nodal signals. This is in complete contrast to the Nodal signal-dependent expression of mixer. These results indicate the complexity of the regulation of mix-type homeobox genes. PMID- 10814509 TI - Poliovirus induces an early impairment of mitochondrial function by inhibiting succinate dehydrogenase activity. AB - Poliovirus infection of COS-1 and T47D cells caused a rapid decrease in total cell respiration, and this was attributed to an inhibition of mitochondrial respiration. The stimulation of mitochondrial respiration by pyruvate plus malate or succinate was impaired in saponin-permeabilised cells. However, this inhibition could be overcome by the addition of N,N,N',N'-tetramethyl-1, 4 phenylenediamine and ascorbate. The activity of succinate dehydrogenase was impaired in parallel with the inhibition of mitochondrial respiration during poliovirus infection. This shows that mitochondrial function is profoundly altered during poliovirus infection and that this occurs primarily through inhibition of electron flow at complex II of the mitochondrial respiratory chain. PMID- 10814510 TI - Putative P-glycoprotein expression in arsenite-resistant Leishmania donovani down regulated by verapamil. AB - Western immunoblots of whole cell lysate and crude membrane extract of an in vitro selected sodium m-arsenite-resistant L. donovani strain revealed a 230-kDa protein identified by an anti-P-glycoprotein (Pgp) antibody. Immunofluorescence microscopy, using the same antibody, detected putative Pgp on resistant parasites. Overexpression of the putative Pgp was down-regulated by verapamil. These results provided, possibly, the first evidence that (i) overexpression of Pgp-like protein occurs in arsenite-resistant Leishmania that are cross-resistant to structurally and functionally unrelated drugs and (ii) verapamil regulates drug sensitivity possibly by down-regulating Pgp expression in the arsenite resistant Leishmania. PMID- 10814511 TI - Increase in phospholipase C-delta1 protein levels in aluminum-treated rat brains. AB - The effect of administration of aluminum to rats on the level of three phospholipase C (PLC) isozymes (beta1, gamma1, and delta1) was assessed in a variety of brain tissues. After exposure to aluminum, a statistically significant increase in malondialdehyde, an index of lipid peroxidation, was observed. In addition, there was a significant reduction in the catalytic activity of low molecular weight phosphotyrosine phosphatase, which loses its activity during oxidative stress. This suggests that oxidative stress is induced in brain tissues exposed to aluminum. The protein level of PLC-delta1, but not that of PLC-beta1 or -gamma1, was significantly increased in brains where oxidative stress had been induced. The total PLC activity in aluminum-treated rat brains was significantly higher than that in control brains. These results suggest that PLC-delta1 protein levels in brain tissues are increased by the induction of oxidative stress, giving an explanation for its up-regulation in Alzheimer's disease. PMID- 10814512 TI - Rho small G-protein-dependent binding of mDia to an Src homology 3 domain containing IRSp53/BAIAP2. AB - mDia1 is a downstream effector of Rho small G protein that is implicated in stress fiber formation and cytokinesis. We isolated an mDia1-binding protein and identified it to be IRSp53/BAIAP2. IRSp53 and BAIAP2 have independently been isolated as a 58/53-kDa protein tyrosine phosphorylated in response to insulin and a BAI1-binding protein, respectively. BAI1 is a brain-specific seven-span transmembrane protein capable of inhibiting angiogenesis. The proline-rich formin homology 1 domain of mDia1 bound the Src homology 3 domain of IRSp53/BAIAP2 in a GTP-Rho-dependent manner. The results suggest that IRSp53/BAIAP2 is a downstream effector of mDia1. PMID- 10814513 TI - Involvement of SPARC in in vitro differentiation of skeletal myoblasts. AB - SPARC (secreted protein acidic and rich in cysteine) is an extracellular Ca(2+) binding glycoprotein associated with the morphogenesis and remodeling of various tissues. Here, involvement of SPARC in the myogenesis of skeletal myoblasts was investigated in vitro. First, the differential expression of SPARC mRNA during the myogenesis was initially identified by a differential display reverse transcription (DDRT)-PCR method. The expression of the SPARC gene was significantly up-regulated during the differentiation of C2C12 mouse myoblasts. Second, the treatment with anti-SPARC antibody almost completely prevented the differentiation of myoblasts. Third, the treatment with EGTA, a Ca(2+) chelator that is known to inhibit the fusion of C2C12 myoblasts, reversibly inhibited the up-regulation of SPARC gene expression. On the other hand, the treatment with A23187, a Ca(2+) ionophore, rapidly and dramatically increased the level of SPARC transcript. Taken together, these results suggest that SPARC may play a critical role(s) in the morphological change of myoblasts, and that the expression of SPARC gene may be controlled by Ca(2+)-dependent pathway in myogenesis. PMID- 10814514 TI - Phosphofructokinase C isozyme from ascites tumor cells: cloning, expression, and properties. AB - The phosphofructokinase C isozyme (PFK-C) from ascites tumor cells has been cloned and characterized to investigate the particular properties of PFK activity in this type of cells. The isolated cDNA encodes a protein of 784 amino acids and 85.5 kDa, whose expression was constant along tumor growth and markedly decreased when cell proliferation stops. The enzyme was functionally expressed in a PFK deficient strain of Saccharomyces cerevisiae and purified to homogeneity. Recombinant PFK-C exhibited the same subunit size as the tumor wild-type isozyme and its steady-state kinetic parameters were similar to those of the form present in normal cells. The regulatory properties of the C isozyme accounted for the lack of fructose-1,6-P(2) activation and the P-enolpyruvate inhibition of PFK activity observed in ascites tumor preparations containing the various isozyme types. Nevertheless, PFK-C binds fructose-1,6-P(2) to an allosteric site as suggested by protection against thermal denaturation. Our results indicate that glucose metabolism in tumor cells is not regulated by a mutant form of PFK-C but by a high level expression of the normal C isozyme. PMID- 10814515 TI - Squamous differentiation downregulates Muc1 mucin in hamster tracheal surface epithelial cell. AB - To investigate whether the squamous differentiation of primary hamster tracheal epithelial cell, which is induced by retinoic acid deficiency or chronic PMA treatment, regulates Muc1 expression, we first produced and characterized a monoclonal antibody against hamster tracheal Muc1 mucin using pGEX-Muc1 fusion protein as an antigen and the changes of Muc1 mucin expression was determined by Western blot. Squamous differentiation downregulated the expression of Muc1 mucin from HTSE cells. The decrease in the immunoreactivity of Muc1 mucin was parallel to the decrease in the immunoreactivity of high molecular weight mucin, which is secreted from HTSE cells. The data from the present study implicate a possible role of Muc1 mucin in squamous differentiation of HTSE cells. PMID- 10814516 TI - Importance of the region including aspartates 57 and 60 of ferredoxin on the electron transfer complex with photosystem I in the cyanobacterium Synechocystis sp. PCC 6803. AB - Ferredoxin reduction by photosystem I has been studied by flash-absorption spectroscopy. Aspartate residues 20, 57, and 60 of ferredoxin were changed to alanine, cysteine, arginine, or lysine. On the one hand, electron transfer from photosystem I to all mutated ferredoxins still occurs on a microsecond time scale, with half-times of ferredoxin reduction mostly conserved compared to wild type ferredoxin. On the other hand, the total amplitude of the fast first-order reduction varies largely when residues 57 or 60 are modified, in apparent relation to the charge modification (neutralized or inverted). Substituting these two residues for lysine or arginine induce strong effects on ferredoxin binding (up to sixfold increase in K(D)), whereas the same substitution on aspartate 20, a spatially related residue, results in moderate effects (maximum twofold increase in K(D)). In addition, double mutations to arginine or lysine were performed on both aspartates 57 and 60. The mutated proteins have a 15- to 20 fold increased K(D) and show strong modifications in the amplitudes of the fast reduction kinetics. These results indicate that the acidic area of ferredoxin including aspartates 57 and 60, located opposite to the C-terminus, is crucial for high affinity interactions with photosystem I. PMID- 10814517 TI - Generation of 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand, in picrotoxinin-administered rat brain. AB - The levels of 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand, and other molecular species of monoacylglycerols in rat brain were examined. In this study, we sacrificed the animals in liquid nitrogen to minimize postmortem changes. We found that rat brain contains 0.23 nmol/g tissue of 2 arachidonoylglycerol, which accounts for 10.5% of the total monoacylglycerol present in this tissue. We next investigated the level of 2-arachidonoylglycerol after in vivo stimulation with picrotoxinin. We found that the level of 2 arachidonoylglycerol was elevated markedly in picrotoxinin-administered rat brain (4- to 6-fold over the control level). Changes in the levels of other molecular species were relatively small or negligible. Several cannabimimetic molecules as well as Delta(9)-tetrahydrocannabinol are known to depress neurotransmission and to exert anticonvulsant activities; endogenous 2-arachidonoylglycerol produced during neural excitation may play a regulatory role in calming the enhanced synaptic transmission. PMID- 10814518 TI - Surface expression of the cystic fibrosis transmembrane conductance regulator mutant DeltaF508 is markedly upregulated by combination treatment with sodium butyrate and low temperature. AB - The DeltaF508 gene mutation prevents delivery of the cystic fibrosis transmembrane conductance regulator (CFTR) to the plasma membrane. The current study examines the biochemical basis for the upregulation of DeltaF508 CFTR expression by sodium butyrate and low temperature. Surface CFTR protein expression was determined by quantitative immunoblot following surface biotinylation and streptavidin extraction. CF gene expression was measured by Northern analysis and CFTR function by forskolin-stimulated (125)I efflux. Butyrate increased DeltaF508 mRNA levels and protein expression but did not increase the biochemical or functional expression of DeltaF508 CFTR at the cell surface. Low temperature increased the biochemical and functional expression of DeltaF508 CFTR at the cell surface but did not increase CFTR mRNA levels. Combining treatments led to a synergistic increase in both DeltaF508 mRNA and surface protein levels that results from the stabilization of CFTR mRNA and protein by low temperature. These findings indicate that surface expression of DeltaF508 CFTR can be markedly enhanced by carefully selected combination agents. PMID- 10814519 TI - Repression of heme oxygenase-1 by hypoxia in vascular endothelial cells. AB - Heme oxygenase 1 (HO-1), a rate-limiting enzyme in heme catabolism, has been reported to be induced by hypoxia. Unexpectedly, here we show that expression of HO-1 mRNA is repressed by hypoxia in primary cultures of human umbilical vein endothelial cells (HUVECs), but is increased by cobalt chloride (CoCl(2)) that is known to mimic hypoxia. Under the culture conditions used, the DNA-binding and transactivation activities of hypoxia-inducible factor 1 were increased in HUVECs by hypoxia or CoCl(2). Therefore, hypoxia and cobalt showed opposing effects on HO-1 mRNA expression, despite activation of hypoxia-inducible factor 1. The half life of HO-1 mRNA was not changed by hypoxia, but was significantly prolonged by CoCl(2). Hypoxia also represses HO-1 mRNA expression in human coronary artery endothelial cells and astrocytes. The repression of HO-1 expression may represent the adaptation to hypoxia in certain cell types. PMID- 10814520 TI - A retroviral vector capable of targeted gene transfer into cells expressing HIV envelope glycoprotein. AB - An expression vector encoding a chimeric envelope protein composed of CD4 and ecotropic retroviral envelope glycoprotein was constructed with the aim of accomplishing targeted gene transfer into HIV-1-infected cells. The chimeric protein was efficiently expressed and transported to the surfaces of various cell types and supported HIV-1 entry into human cells. A packaging cell line producing retroviral vectors carrying chimeric envelope proteins was then established. The vector particles produced were shown to be capable of specific gene transfer into human cells expressing HIV envelope glycoprotein. Blocking experiments confirmed that the vector particles entered the cells via an interaction between the chimeric and HIV envelope proteins. This targeting vector may thus be a useful tool with which to develop effective gene therapies against HIV infection. PMID- 10814521 TI - Expression of isolated C-type carbohydrate recognition domains. AB - A galactose-binding lectin from the venom of the snake Trimeresurus stejnegeri consists of isolated carbohydrate recognition domains, belonging to group VII of the C-type animal lectins. As a first step toward determining the tertiary structure of the galactose-specific lectin, we produced the lectin in Escherichia coli. By in vitro refolding and affinity chromatography, modest amounts (8 mg/liter) of active recombinant proteins were obtained. The recombinant protein was homogeneous, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. Its amino acid sequence without the initiated methionine at the N-terminus was also characterized by mass spectrometry. The data of hemagglutination and enzyme-linked lectin binding assays demonstrated that the recombinant lectin showed similar sugar-binding activity as the native protein. In addition, fluorescence spectroscopy and circular dichroism also showed obviously their structural similarity. PMID- 10814522 TI - Mouse smad8 phosphorylation downstream of BMP receptors ALK-2, ALK-3, and ALK-6 induces its association with Smad4 and transcriptional activity. AB - Smads are intracellular signaling mediators for TGF-beta superfamily. Smad1 and Smad5 are activated by BMP receptors. Here, we have cloned mouse Smad8 and functionally characterized its ability to transduce signals from BMP receptors. Constitutively active BMP type I receptors, ALK-3 and ALK-6, as well as ALK-2, were phosphorylated Smad8 and induced Smad8 interaction with Smad4. Nuclear translocation of Smad8 was stimulated by constitutively active BMP type I receptors. In contrast, constitutively active TGF-beta type I receptor, ALK-5, did not exhibit any action on Smad8. Smad8 and Smad4 cooperatively induced the promoter of Xvent2, a homeobox gene that responds specifically to BMP signaling. Dominant-negative Smad8 was shown to inhibit the increase of alkaline phosphatase activity induced by BMP-2 on pluripotent mesenchymal C3H10T1/2 and myoblastic C2C12 cell lines. The presence of Smad8 mRNA in mouse calvaria cells and osteoblasts suggests a role of Smad8 in the osteoblast differentiation and maturation. PMID- 10814523 TI - Compactin and simvastatin, but not pravastatin, induce bone morphogenetic protein 2 in human osteosarcoma cells. AB - Bone morphogenetic protein (BMP)-2, a member of the BMP family, plays an important role in osteoblast differentiation and bone formation. To discover small molecules that induce BMP-2, a luciferase reporter vector containing the 5' flanking promoter region of the human BMP-2 gene was constructed and transfected into human osteosarcoma (HOS) cells. By the screening of an in-house natural product library with stably transfected HOS cells, a fungal metabolite, compactin, known as an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, was isolated. The stimulation of the promoter activity by compactin seemed to be specific for BMP-2 gene in HOS cells, since it had little effect on BMP-4 or SV40 promoter activity and the stimulation was not observed in Chinese hamster ovary (CHO) cells. RT-PCR analysis and alkaline phosphatase assay revealed that compactin induced an increase in the expression of BMP-2 mRNA and protein. Like compactin, simvastatin also activated the BMP-2 promoter, whereas pravastatin did not. The statin-mediated activation of BMP-2 promoter was completely inhibited by the downstream metabolite of HMG-CoA reductase, mevalonate, indicating that the activation was a result of the inhibition of the enzyme. These results suggest that statins, if they are selectively targeted to bone, have beneficial effects in the treatment of osteoporosis or bone fracture. PMID- 10814524 TI - Isolation of two novel genes, DSCR5 and DSCR6, from Down syndrome critical region on human chromosome 21q22.2. AB - We have isolated two novel genes, designated DSCR5 and DSCR6, from the Down syndrome critical region (DSCR) on chromosome 21q22.2 which has been defined as minimal overlapping region of partial trisomy 21 patients and located between t(4;21) break point and ERG (approximately 1.6 Mb). DSCR5 and DSCR6 genes consist of 6 and 5 exons, respectively. Alternative use of transcription start sites and alternative splicing events produce different RNA species and proteins from both genes. Three different transcripts of DSCR5 gene encode three putative transmembrane proteins of 158, 134, and 108 amino acids, while 4 different transcripts of DSCR6 gene encode two forms of proteins with 190 and 106 amino acids. The DSCR5 gene is expressed in various human tissues examined, whereas the DSCR6 gene is expressed only in limited tissues at low level. Both DSCR5 and DSCR6 genes are candidates for the pathogenesis of Down syndrome, although the function of these genes remains to be elucidated. PMID- 10814525 TI - Evidence of hepatic endogenous hydrogen peroxide in bile of selenium-deficient rats. AB - Hepatic endogenous hydrogen peroxide (H(2)O(2)) in bile of selenium-deficient rats (SeD) was for the first time found using the electron spin resonance (ESR) spin-trap technique, and the relationship between glutathione peroxidase (GPX) activity and H(2)O(2) amount is discussed. Normal rats and four groups of rats fed a selenium-deficient diet with different feeding periods were examined. The results showed that the GPX activity decreased depending on the feeding period with the selenium-deficient diet and that the hepatic endogenous H(2)O(2) amount in the bile of the rats fed the selenium-deficient diet for the longest period (a week before birth to 8 weeks old) was drastically higher than those in other groups of rats (P < 0.005). We found that generation of H(2)O(2) due to the decrease in the GPX activity has a threshold value. The results suggest that an exposure to selenium deficiency for long term will cause oxidative stress. PMID- 10814526 TI - Differential regulation of leptin receptor expression by insulin and leptin in neuroblastoma cells. AB - Leptin exerts its effects by interacting with specific membrane receptors (Ob-R). We studied the exact localization of long intracellular domain form (Ob-Rb) in human brain. In addition, we analyzed the regulatory features of Ob-Rb expression in two neuroblastoma cell lines. The Ob-Rb mRNAs were abundant in putamen, frontal lobe, medulla, cerebral cortex, cerebellum, thalamus, hippocampus, corpus callosum, caudate nucleus, and amygdala, indicating that Ob-Rb transcripts are expressed differently from that of other Ob-R isoforms. In SK-N-MC cells, the expression of Ob-Rb mRNA was induced by increasing doses of insulin, and the maximum amount of mRNA expression was 9.4-fold higher in the presence of insulin (100 nM for 24 h), compared to the absence of insulin. In IMR32 cells, the transcripts were increased 4.0-fold when cells were incubated with 1 nM of insulin for 48 h. In contrast, Ob-Rb expression in IMR32 cells decreased to 18% of control following a 24-h incubation period with 50 ng/mL of leptin, compared to incubation in the absence of leptin. These results indicate that expression of Ob-Rb is differentially regulated by inhibitory signals of energy balance in neuroblastoma cells. The identification of the novel regulatory mechanisms involving the Ob-Rb isoform by insulin and leptin now makes it possible to elucidate the underlying mechanisms involving increased food intake and uncontrolled energy balance associated with leptin resistance in obese individuals. PMID- 10814527 TI - Insulin sensitizer, troglitazone, directly inhibits aromatase activity in human ovarian granulosa cells. AB - Ovarian granulosa cells synthesize estrogens from androgens, which are catalyzed by aromatase cytochrome P450 (P450arom). Troglitazone (Tro), one of the insulin sensitizing compounds, thiazolidinediones (TZDs), is a ligand for peroxisome proliferator activated receptor gamma (PPARgamma) and is effective in the treatment of both non-insulin-dependent diabetes mellitus (NIDDM) as well as polycystic ovary syndrome (PCOS). PPARgamma exerts a transcriptional activity as a PPARgamma:RXR heterodimer. In this study, we investigated the effects of Tro and/or RXR ligand, LG100268 (LG) on the aromatase activity in cultured human ovarian granulosa cells obtained from patients who underwent in vitro fertilization. Human ovarian granulosa cells expressed PPARgamma mRNA assessed by RT-PCR. The treatment of the granulosa cells with Tro for 24 h resulted in a dramatic inhibition of the aromatase activity in a dose-dependent manner. While the treatment with LG alone also inhibited the aromatase activity, the combined treatment with both Tro and LG caused a much more reduction in the aromatase activity. The changes in the aromatase activity by Tro and/or LG were associated with comparable changes in P450arom mRNA assessed by RT-PCR. These results suggest that Tro directly inhibit the aromatase activity in human granulosa cells probably via nuclear receptor system PPARgamma:RXR heterodimer. The findings may provide a biochemical basis for the decrease in the blood concentrations of estrogens which is observed after the in vivo administration of Tro and may also possibly be useful as a novel therapy for estrogen-dependent diseases. PMID- 10814528 TI - Translational defects of Escherichia coli mutants deficient in the Um(2552) 23S ribosomal RNA methyltransferase RrmJ/FTSJ. AB - We recently identified RrmJ (alias FtsJ), the first encoded protein of the rrmJ hflB heat shock operon, as an Um(2552) methyltransferase of the 23S rRNA. We now report that the rrmJ-deficient strain exhibits growth and translational defects compared to the wild-type strain. Growth rates of the rrmJ mutant are decreased at both low and high temperatures. Protein synthesis activity is reduced up to 65% when S(30) rrmJ mutant extracts are tested in a coupled in vitro transcription/translation assay. In vitro methylation of these extracts by RrmJ partially restores protein synthesis activity. Polysome profile analysis of the rrmJ strain reveals an increase in the proportion of free 30S and 50S subunits at both 30 and 42 degrees C. These results suggest that the RrmJ-catalyzed methylation of Um(2552) in 23S RNA strengthens ribosomal subunit interactions, increases protein synthesis activity, and improves cell growth rates even at non heat shock temperatures. PMID- 10814529 TI - Adaptor gamma ear homology domain conserved in gamma-adaptin and GGA proteins that interact with gamma-synergin. AB - We identified a novel family of proteins that have a VHS domain and an AGEH (adaptor gamma ear homology) domain that is homologous to the ear domain of the gamma-adaptin subunit of the AP-1 clathrin adaptor. When overexpressed, the proteins, called GGA1, GGA2, and GGA3, localized to the trans-Golgi network (TGN) and often caused fragmentation and vacuolation of the compartment. Yeast two hybrid analysis showed that the AGEH domains of the GGA proteins as well as those of gamma-adaptins are able to interact with gamma-synergin, which was previously shown to localized in the TGN region and interact with gamma-adaptin. Furthermore, gamma-synergin and either of the GGA proteins coexpressed were colocalized in the TGN region. These results suggest that the GGA proteins regulate the function of the TGN or membrane trafficking from this compartment and that the AGEH domains of GGAs and gamma-adaptins, like the ear domain of alpha-adaptin, are involved in interaction with molecules that modulate their functions. PMID- 10814530 TI - Melanin binds reversibly to thermostable DNA polymerase and inhibits its activity. AB - We found that both RNA and cDNA preparations derived from melanocytes contain a RT-PCR inhibitor that copurified with nucleic acids. Investigation of the candidate inhibitor melanin revealed that it potently blocks PCR at concentrations below 200 ng/ml, whereas 100 microg/ml melanin was required to inhibit reverse transcription. Melanin and thermostable DNA polymerase preferentially formed a distinct complex with reduced migration velocity as compared to pure polymerase in nondenaturating polyacrylamide gel electrophoresis. The inhibition of the enzyme by melanin could be reversed by diluting solutions of preformed complexes or by adding excess amounts of other proteins such as bovine serum albumin or dry milk. Our findings demonstrate that melanin is a potent inhibitor of thermostable DNA polymerase in vitro and that the inhibitory effect is conferred by a direct and reversible polymerase-melanin interaction. PMID- 10814531 TI - DNA cleavage by a Chromium(III) complex. AB - A new Cr(III) complex with the empirical formula [Cr(Schiff base) (H(2)O)(2)]ClO(4), where the Schiff base is 2, 3-bis?[(2-hydroxy-4-diethylamino) (phenyl) (methylene)]amino?2-butenedinitrile has been synthesized and characterized spectroscopically. Binding of this complex to DNA has been studied using UV-visible spectroscopy. The complex has been found to bind to the major groove of DNA with a binding constant, K = (1.3 +/- 0.2) x 10(3) M(-1). The induced CD spectrum of the complex in the presence of DNA is also indicative of major groove binding. Gel electrophoresis of plasmid DNA in the presence of the complex shows that the complex brings about nicking of the DNA. PMID- 10814532 TI - Seasonal adaptation modulates the expression of the protein kinase CK2 beta subunit gene in the carp. AB - Carp fish seasonal acclimatization induces a cyclical transcriptional modulation of several genes. Its most dramatic expression results in a concomitant structural rearrangement of the nucleolar components that phenotypically represents profound shifts in the level of ribosomal RNA synthesis. In connection with the recent studies that implicate CK2 in the control of rRNA synthesis in vertebrates, we characterized the cDNA of carp protein kinase CK2beta subunit and assessed its transcriptional behavior in winter- and summer-acclimatized fish. We found a remarkable differential gene expression of CK2beta subunit between summer and winter-acclimatized carp which correlates with the modulatory pattern observed in rRNA transcription. PMID- 10814533 TI - An air-liquid interface promotes the differentiation of gastric surface mucous cells (GSM06) in culture. AB - The gastric surface epithelium is situated at an air-liquid interface because the luminal surface of the alimentary tract is in continuity with the air phase. However, the effects of this microenvironment on the gastric epithelium remain unclear. The aim of this study was to clarify the effects of an air-liquid interface on gastric epithelial cell biology. Gastric surface mucous cells (GSM06) were cultured at an air-liquid interface. Cultured cells were examined by histology, histochemistry, and transmission electron microscopy. When the cells were cultured at an air-liquid interface, the surface cells on the collagen gel became tall columnar and secreted periodic acid-Shiff-positive substances at the apical surface. These cells indicated many mucous granules in the apical cytoplasm and organized the basal lamina at the contact side with the gel. In contrast, under immersed condition, the surface cells showed immature features. This is the first report of an air-liquid interface promoting the differentiation of gastric surface mucous cells in a reconstruction culture of the gastric surface epithelial layer, suggesting that an air-liquid interface may function as a crucial luminal factor to maintain the homeostasis of gastric mucosa. PMID- 10814534 TI - Apoptosis induced by hydrogen peroxide under serum deprivation and its inhibition by antisense c-jun in F-MEL cells. AB - Under serum deprivation F-MEL cells die by apoptosis. We previously showed that apoptosis induced by serum deprivation was suppressed by inhibition of c-jun expression using antisense c-jun transfected cell line, c-junAS. To elucidate the underlying mechanisms we examined the species which is responsible for apoptosis under serum deprivation. When catalase and N-acetyl-L-cysteine (NAC) were included in the medium, cell death under serum deprivation was effectively suppressed in F-MEL cells. Intracellular generation of hydrogen peroxide (H(2)O(2)) was also detected under serum deprivation in parental F-MEL cells, but it was suppressed in c-junAS (+) cells, in which antisense c-jun was expressed and c-Jun protein expression was inhibited as shown by Western blot. When H(2)O(2) was directly applied to F-MEL cells at 3 mM, apoptotic cell death was induced, whereas it was suppressed in c-junAS (+) cells. Induction of apoptosis by H(2)O(2) and its inhibition by antisense c-jun was confirmed by detection of internucleosomal fragmentation of DNA, TdT-mediated dUTP nick end labeling (TUNEL)-positive cells and morphological alteration of nuclei. These results indicate that apoptosis induced by serum deprivation in F-MEL cells is mediated by H(2)O(2) and c-jun expression is essential to apoptosis induced by H(2)O(2) in F-MEL cells. PMID- 10814535 TI - The insulin-dependent glucose transporter isoform 4 is expressed in bovine blastocysts. AB - We have investigated the expression of two glucose transporter isoforms, Glut1 and 4, in 14- and 16-day-old bovine blastocysts (d14, d16) using RT-PCR, competitive RT-PCR and in situ hybridization. The blastocysts were grown in vivo or had been produced in vitro. Glut1 mRNA was detected in all blastocysts studied, Glut4 in all d14 blastocysts, but only in a few d16 blastocysts. Glut4 mRNA was localized in trophoblast and endoderm cells. Glut1 mRNA increased from d14 to d16 while Glut4 transcription was down-regulated in d16 blastocysts. The mRNA amounts varied between 0.8 to 23 pg and 3.9 to 65 fg per 100 ng embryonic RNA for Glut1 and Glut4, respectively, displaying a 100- to 1500-fold lower expression of Glut4 compared with Glut1 during blastocyst elongation. This is the first report on the expression of the insulin-sensitive Glut4 isoform in mammalian preimplantation embryos. PMID- 10814536 TI - Cloning of a novel G-protein-coupled receptor from the sea anemone nervous system. AB - We describe the cloning and analysis of genomic and cDNA copies of a gene from sea anemones that encodes a new member of the G-protein-coupled receptor family. The receptor shows similarity to previously described receptors for biogenic amines such as adrenaline, serotonin, and octopamine, as well as a variety of small molecule agonists and peptides, although we have been unable to determine which ligand is the natural agonist. Antibodies generated against the recombinant receptor protein identify a single protein with a molecular weight of 66 kDa in membrane preparations. Immunofluorescence studies using the same antibody have enabled localization of the receptor in the nervous system. Western blotting and RT-PCR analysis reveal that a homologue of this receptor is expressed in jellyfish and soft coral. We suggest that the receptor plays a role in neurotransmission in the sea anemone and other members of the phylum Cnidaria. PMID- 10814537 TI - Effect of halothane on the oligomerization of the sarcoplasmic reticulum Ca(2+) ATPase. AB - The exact molecular mechanism of inhalational anesthetics remains obscure. Since the enzyme activity of the sarcoplasmic reticulum Ca(2+)-ATPase from skeletal muscle fibres is modified by halothane and because protein-protein interactions play an important role in the regulation of Ca(2+)-regulatory proteins, we investigated the effect of this volatile drug on the oligomerization of the fast twitch Ca(2+)-ATPase. Using electrophoretic separation following incubation with halothane, increases in relative molecular mass were determined by immunoblotting with a monoclonal antibody to the SERCA1 isoform of the Ca(2+)-ATPase. Distinct drug-induced decreases in electrophoretic mobility indicated oligomerization of the native Ca(2+)-pump by halothane, comparable to crosslinking-mediated formation of homo-tetramers. Determination of the effect of halothane on enzyme activity suggested that halothane-mediated protein aggregation triggers a partial inhibition of Ca(2+)-pump units. Thus, halothane appears to exert its action via specific peptide binding sites and not indirectly by lipid perturbation. These findings support the protein theory of anesthetic action. PMID- 10814538 TI - Rat histidine decarboxylase is a substrate for m-calpain in vitro. AB - We have followed the in vitro degradation of rat histidine decarboxylase in a reconstituted system, containing only rat histidine decarboxylase (obtained by in vitro transcription and translation), calcium ions in the millimolar range of concentrations, and m-calpain. Under the experimental conditions used, m-calpain quickly and efficiently degraded rat histidine decarboxylase, giving rise to a major proteolytic band of 29 kDa. In a conventional in vitro degradation system containing rabbit reticulocytes supplemented with calcium ions, there was also an intense proteolysis of rat histidine decarboxylase, strongly inhibited in the presence of calpeptin, a highly specific calpain inhibitor. PMID- 10814539 TI - Xeroderma pigmentosum complementation group A protein acts as a processivity factor. AB - We have previously shown that endonucleases present in a protein complex, which has specificity for cyclobutane pyrimidine dimers, locate sites of damage in DNA by a processive mechanism of action in normal human lymphoblastoid cells. In contrast, the endonucleases present in this complex from xeroderma pigmentosum complementation group A (XPA) cells locate damage sites by a distributive or significantly less processive mechanism. Since the XPA protein has been shown to be responsible for the DNA repair defect in XPA cells, this protein was examined for involvement in the mechanism of target site location of these endonucleases. A recombinant XPA protein, produced by expression of the normal XPA cDNA in E. coli, was isolated and purified. The results show that the recombinant XPA protein was able to correct the defect in ability of the XPA endonucleases to act by a processive mechanism of action on UVC irradiated DNA. These studies indicate that the XPA protein, in addition to a role in damage recognition or damage verification, may function as a processivity factor. PMID- 10814540 TI - NADH-ferric reductase activity associated with dihydropteridine reductase. AB - In mammals dietary ferric iron is reduced to ferrous iron for more efficient absorption by the intestine. Analysis of a pig duodenal membrane fraction revealed two NADH-dependent ferric reductase activities, one associated with a b type cytochrome and the other not. Purification and characterization of the non cytochrome ferric reductase identified a 31 kDa protein. MALDI-MS analysis and amino acid sequencing identified the ferric reductase as being related to the 26 kDa liver NADH-dependent quinoid dihydropteridine reductase (DHPR). The NADH dependent DHPR ferric reductase activity was found to be pteridine-independent since exhaustive dialysis did not reduce activity and heat-inactivation destroyed activity. In intestinal Caco-2 cells, DHPR mRNA levels were found to be regulated by iron. Thus, DHPR appears to be a dual function enzyme, a NADH-dependent dihydopteridine reductase and an iron-regulated, NADH-dependent, pteridine independent ferric reductase. PMID- 10814541 TI - A possible interaction of thioredoxin with VDUP1 in HeLa cells detected in a yeast two-hybrid system. AB - Human thioredoxin (hTrx), a small ubiquitous protein with strong reducing potential, has multiple biological functions, including signal transduction and regulation of the activity of transcription factors. hTrx expression is enhanced in HPV-transformed cancer cells; however, the role of hTrx in the malignant cells is not fully understood. We employed a yeast two-hybrid system to search for proteins that bind to hTrx in HeLa cells, a type of HPV-transformed human cervical cancer cell. In a screen of 1.62 x 10(6) yeast cotransformed with a HeLa cDNA library and an hTrx vector, 13 clones were identified as candidates for hTrx binding proteins. Among them, 3 clones were found to code in frame for the carboxyl-terminal portion of VDUP1 protein, lacking at most the first 155 residues from the start codon. A reconstructed clone carrying the full-length VDUP1 coding sequence also showed the ability to bind to an hTrx fusion protein. Loss of interaction between VDUP1 and hTrx was observed either when two cysteines (Cys 32 and 35) in hTrx were substituted by serines or when the deletion in VDUP1 was extended from amino acid position 155 to 225 or beyond. The 71-mer peptide fragment (position 155-225) of VDUP-1 alone did not bind to hTrx. PMID- 10814542 TI - Structural and functional role of cysteinyl residues in tobacco acetolactate synthase. AB - Acetolactate synthase (ALS) is the common enzyme in the biosynthesis of valine, leucine, and isoleucine. The role of four cysteinyl residues in tobacco ALS was determined using site-directed mutagenesis and cysteine-specific cleavage. The C411A mutation abolished the enzymatic activity, as well as the binding affinity for the cofactor FAD. The activation constant of C411S for FAD is approximately 50-fold higher than that of wALS. The C607S mutation did not significantly affect the kinetic parameters. The IC(50) values of C411S and C607S for ALS-inhibiting herbicides are not much different from those of wALS. Two mutants, C163S and C309S, are labile and readily degraded to peptide fragments. The treatment of wALS with 2-nitro-5-thiocyanobenzoic acid, specific for cleavage of the N terminal side of cysteine, yielded three peptides of 37.0, 22. 0, and 7.0 kDa. This fragmentation pattern is consistent with that deduced from the amino acid sequence of tobacco ALS, assuming the disulfide bond between Cys163 and Cys309. These results suggest that Cys411 is involved in the binding of FAD and that the intrachain disulfide bond between Cys163 and Cys309 plays a key role in maintaining the correct conformation of tobacco ALS. PMID- 10814543 TI - Endothelin-1 chronically inhibits Na/H exchanger-3 in ET(B)-overexpressing OKP cells. AB - Endothelin-1 (ET-1) acutely increases Na/H antiporter activity in OKPET(B)6 cells, an opossum kidney proximal tubule cell line transfected with ET(B) receptor cDNA. The purpose of the present study was to examine the chronic effect of ET-1 on Na/H antiporter activity in OKP cells and to examine whether Na/H exchanger (NHE)-3 mRNA and protein abundance are regulated by ET-1. Quiescent OKPET(B)6 cells were treated with 10 nM ET-1 for 3, 6 or 24 h and Na/H antiporter activity was assayed. The Na/H antiporter activity in 3-h ET-1-treated cells was not different from controls. However, Na/H antiporter activity was significantly decreased by 29% at 6 h and 72% at 24 h. The effect of ET-1 on Na/H antiporter activity was blocked by BQ788, an ET(B) receptor antagonist, but not BQ123, an ET(A) receptor antagonist. The NHE-3 mRNA abundance in ET-1-treated cells was not different from controls at 3 h. However, there was a significant decrease in NHE 3 mRNA abundance at 6 and 24 h. There was also a significant decrease in NHE-3 protein abundance at 6 and 24 h. In summary, ET-1 chronically inhibits NHE-3 in OKPET(B)6 cells. PMID- 10814544 TI - Roles of cytosolic phospholipase A(2) and platelet-activating factor receptor in the Ca-induced biosynthesis of PAF. AB - Casein-elicited peritoneal exudate cells (PEC), mainly consisted of neutrophils, were collected from platelet-activating factor receptor-knock-out (PAFR-KO), cytosolic phospholipase A(2) knock-out (cPLA(2)-KO), and wild-type (WT) mice. After stimulation of PEC with calcium ionophore A 23187, PAF levels were measured by radio-ligand binding assay using receptor-rich membrane fraction prepared from the PAF receptor transgenic mice. We found that the level of PAF production by PEC was not different between WT and PAFR-KO mice. On the other hand, cPLA(2)-KO mice were deficient in the PAF production. These results provide the direct evidence while cPLA(2) is essential in the production of PAF, PAF receptor deficiency has little effect on the PAF production. PMID- 10814545 TI - Nucleotide binding and sulfation catalyzed by phenol sulfotransferase. AB - The sulfation of a nucleotide is an indispensable step for the sulfuryl group transfer in a biological system. The product and cosubstrate of sulfotransferase in physiological condition are adenosine 3',5'-bisphosphate (PAP) and 3'-phospho adenosine 5'-phosphosulfate (PAPS), respectively. We find that ribose and adenine, two major parts of the adenosine nucleotide, bind tightly to phenol sulfotransferase (PST) separately, and various nucleotides also bind tightly to PST. We determine the dissociation constants of a variety of nucleotides and examine their potential as cofactors or cosubstrates of PST. Using 4-nitrophenyl sulfate as the sulfuryl group donor, three nucleotides, adenosine 5' monophosphate (AMP), adenosine 2',5'-bisphosphate (2',5'-PAP), and adenosine 2':3'-cyclic phosphate 5'-phosphate (2':3'-cyclic PAP), are shown here for the first time to be sulfated at 5'-phopho position by a PST catalyzed reaction. Spectrophotometry, HPLC, and (31)P NMR are used to determine the activity of PST and identify the sulfated nucleotides. The V(max) of PST and K(m) of these nucleotides are determined when they are used as cofactors or cosubstrates for the sulfuryl group transfer. The existence and possible physiological significance of these newly reported binding and sulfation of nucleotides by PST in biology is yet to be discovered. PMID- 10814546 TI - Volitional ethanol consumption affects overall serotonin metabolism in Syrian golden hamsters (Mesocricetus auratus). AB - Methods were established for the determination of serotonin (5-HT)(1) metabolites 5-hydroxyindole-3-acetic acid (5-HIAA) and 5-hydroxytryptophol (5-HTOL) in the urine of Syrian golden hamsters (Mesocricetus auratus) and used to study the effect of volitional ethanol consumption on overall 5-HT metabolism in this ethanol-preferring rodent. The basal levels of 5-HIAA and 5-HTOL in 24-h urine of ethanol-naive hamsters were 300 +/- 101 and 4.96 +/- 1. 06 nmol (n = 8), respectively. Given free choice between water and a 15% ethanol solution, these hamsters chose to consume increasing amounts of ethanol. The increase was accompanied by a concomitant decrease in urine 5-HIAA and increase in urine 5 HTOL, indicating that volitional ethanol intake diverted part of the 5-HT metabolic flux from an oxidative into a reductive pathway. In a separate experiment, the amounts of ethanol consumed by and blood ethanol concentrations attained in ethanol-drinking golden hamsters were determined at 5 different time intervals between 6 PM and 7 AM when most feeding activities occurred. Except in the first hour after lights were turned off, ethanol was consumed at a relatively even pace throughout the night (2-3 g/kg/3 h) and blood ethanol levels were maintained at the low mM range which rarely exceeded 2 mM. These results suggest that the biochemical pathway that catalyzes 5-HT metabolism is extremely sensitive to ethanol and can play an important role in mediating the reported clinically beneficial action of a low concentration of ethanol during alcohol detoxification. PMID- 10814547 TI - PCR cloning and baculovirus expression of human lactoperoxidase and myeloperoxidase. AB - Lactoperoxidase (LPO) and myeloperoxidase (MPO) have been identified previously in human milk. These peroxidases have antimicrobial activity and presumably contribute to the protective functions of milk. In this study, we amplified genes encoding LPO and MPO from human mammary gland cDNA by the polymerase chain reaction (PCR). These genes were expressed in a baculovirus-insect cell system. Peroxidase activity was observed in the culture supernatant of Tricoplusia ni cells infected with the recombinant viruses and the levels increased upon addition of delta-aminolevulinic acid. Purified recombinant human LPO and MPO, both with a molecular mass of about 80 kDa, showed properties similar to bovine LPO and human MPO, respectively, in terms of absorption spectrum, sensitivity to dapsone, specificity for chloride ions, and reactivity with anti-bovine LPO or anti-MPO antibodies. Our data suggest that this expression system is useful for studying the catalytic mechanism and biological significance of these human peroxidases. PMID- 10814548 TI - Complete sequences of two highly divergent european isolates of TT virus. AB - TT virus is a virus distantly related to the Circoviridae family. We report here the complete genome characterization of two European human isolates (T3PB and TUPB) using a new and simple protocol for sequencing GC-rich genomic regions. Sequence analysis confirmed the existence of two major ORFs, of a CAV-like VP2 motif in ORF2 and of potential stem-loop structures in non-coding regions. Phylogenetic analyses based on complete genomic sequences of human isolates suggested that three different lineages exist at least. The first lineage includes genotypes 1, 2, and 3, and two other lineages include viruses related to the Japanese SANBAN and to the North American TUS01 isolates respectively. Sequence comparison made it possible to assign strain T3PB to genotype 3, and strain TUPB to the TUS01 group. Consequently, this study reports the first full length sequence of a genotype 3 isolate and demonstrates that viruses belonging to the TUS01 lineage are present in the Old Word. PMID- 10814549 TI - A physiological toxicokinetic model for exogenous and endogenous ethylene and ethylene oxide in rat, mouse, and human: formation of 2-hydroxyethyl adducts with hemoglobin and DNA. AB - Ethylene (ET) is a gaseous olefin of considerable industrial importance. It is also ubiquitous in the environment and is produced in plants, mammals, and humans. Uptake of exogenous ET occurs via inhalation. ET is biotransformed to ethylene oxide (EO), which is also an important volatile industrial chemical. This epoxide forms hydroxyethyl adducts with macromolecules such as hemoglobin and DNA and is mutagenic in vivo and in vitro and carcinogenic in experimental animals. It is metabolically eliminated by epoxide hydrolase and glutathione S transferase and a small fraction is exhaled unchanged. To estimate the body burden of EO in rodents and human resulting from exposures to EO and ET, we developed a physiological toxicokinetic model. It describes uptake of ET and EO following inhalation and intraperitoneal administration, endogenous production of ET, enzyme-mediated oxidation of ET to EO, bioavailability of EO, EO metabolism, and formation of 2-hydroxyethyl adducts of hemoglobin and DNA. The model includes compartments representing arterial, venous, and pulmonary blood, liver, muscle, fat, and richly perfused tissues. Partition coefficients and metabolic parameters were derived from experimental data or published values. Model simulations were compared with a series of data collected in rodents or humans. The model describes well the uptake, elimination, and endogenous production of ET in all three species. Simulations of EO concentrations in blood and exhaled air of rodents and humans exposed to EO or ET were in good agreement with measured data. Using published rate constants for the formation of 2-hydroxyethyl adducts with hemoglobin and DNA, adduct levels were predicted and compared with values reported. In humans, predicted hemoglobin adducts resulting from exposure to EO or ET are in agreement with measured values. In rodents, simulated and measured DNA adduct levels agreed generally well, but hemoglobin adducts were underpredicted by a factor of 2 to 3. Obviously, there are inconsistencies between measured DNA and hemoglobin adduct levels. PMID- 10814550 TI - The role of cholinergic and noncholinergic mechanisms in the cardiorespiratory failure produced by N-methylcarbamate cholinesterase inhibitors in rabbits. AB - We investigated the relative contribution of several cardiorespiratory components to acute lethality produced by N-methylcarbamate cholinesterase (ChE) inhibitors physostigmine, 2-sec-butylphenyl methylcarbamate (BPMC), and 2-isopropoxyphenyl methylcarbamate (PHC) in halothane-anesthetized rabbits. Intravenous injection of these compounds produced dose-dependent pressor and/or depressor responses related to each compound. A lethal dose of physostigmine resulted in cardiovascular collapse after a pressor response. That of PHC produced cardiovascular collapse after biphasic effects on blood pressure, a transient decrease followed by an increase. Unlike these compounds, BPMC elicited a rapidly developing depressor response followed by cardiovascular collapse. Artificial ventilation prevented cardiovascular collapse and lethal actions to physostigmine and PHC, but not BPMC. A degree of acute lethality to physostigmine and PHC depended on their anti-ChE activity, whereas BPMC exhibited a low degree of lethality relative to its anti-ChE activity. While the pressor response to physostigmine and PHC was ascribed to an atropine-sensitive increase in cardiac contractility, the depressor response to PHC and BPMC was attributed to an atropine-insensitive decrease in cardiac contractility and/or vascular resistance. Similar to the order for eliciting the depressor response in vivo, all three compounds inhibited contraction of the isolated cardiac and aortic smooth muscles with the order of their inhibition in terms of anti-ChE activity, i.e., BPMC > PHC > physostigmine. Thus, the primary cause of death with physostigmine and PHC is respiratory arrest subsequent to ChE inhibition, whereas BPMC exhibiting the low degree of lethality causes cardiovascular collapse mediated through direct inhibitory effects on cardiac and vascular smooth muscle contraction. PMID- 10814551 TI - Pharmacological analysis of noncholinergic action of 2-sec-butylphenyl N methylcarbamate insecticide on the isolated rabbit aorta. AB - We investigated noncholinergic actions of 2-sec-butylphenyl N-methylcarbamate (BPMC) on the isolated rabbit thoracic aorta to help determine the mechanisms responsible for its unique toxicological properties, which are characterized by cardiovascular collapse and low lethality compared to its anticholinesterase (anti-ChE) activity. BPMC inhibited K(+)-induced contraction more effectively than it did norepinephrine (NE)-induced contraction. The inhibitory effect on K(+)-induced contraction was not altered by changing the external K(+) concentration, but it was decreased by adding an L-type Ca(2+) channel agonist, BAY K 8644. Simultaneous measurement of tension and cytosolic Ca(2+) levels elevated by K(+) stimulation revealed that BPMC decreased the Ca(2+) levels prior to and parallel to the tension. The magnitude of the inhibitory effect on Ca(2+) levels was increased by treating BPMC before Ca(2+) application in the depolarized preparation. However, BPMC did not inhibit caffeine- or NE-induced transient contraction in Ca(2+)-free medium. On the other hand, BPMC produced tonic contraction in the resting aorta. The contraction to BPMC did not develop after removing the adventitia. The contraction was inhibited by phentolamine or guanethidine but not by atropine or tetrodotoxin. These results suggest that BPMC inhibits depolarization- or agonist-induced contraction by inhibiting Ca(2+) entry through L-type Ca(2+) channels, whereas it produces vascular contraction in the resting state by releasing NE from sympathetic nerve terminals. These apparently opposing mechanisms may contribute to the unique noncholinergic toxicological properties of BPMC. PMID- 10814552 TI - Biochemical basis for the extreme sensitivity of turkeys to aflatoxin B(1). AB - Poultry are the most susceptible food animal species to the toxic effects of the mycotoxin aflatoxin B(1) (AFB(1)). Feed contaminated with even small amounts of AFB(1) results in significant adverse health effects in poultry. The purpose of this study was to explain the biochemical mechanism(s) for this extreme sensitivity. We measured microsomal activation of AFB(1) to the AFB(1)-8,9 epoxide (AFBO), the putative toxic intermediate, as well as cytosolic glutathione S-transferase (GST)-mediated detoxification of AFBO, in addition to other hepatic phase I and phase II enzyme activities, in 3-week-old male Oorlop strain turkeys. Liver microsomes prepared from these turkeys activated AFB(1) in vitro with an apparent K(m) of 109 microM and a V(max) of 1.25 nmol/mg/min. Preliminary evidence for the involvement of cytochromes P450 (CYP) 1A2 and, to a lesser extent, 3A4 for AFB(1) activation was assessed by the use of specific mammalian CYP inhibitors. The possible presence of avian orthologues of these CYPs was supported by activity toward ethoxyresorufin and nifedipine, as well as by Western immunoblotting using antibodies to human CYPs. Cytosol prepared from turkey livers exhibited GST-mediated conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and 3,4-dichloronitrobenzene (DCNB), but at a much lower rate than that observed in other species. Western immunoblotting indicated the presence of alpha and sigma class GSTs and another AFB(1)-detoxifying enzyme, AFB(1)-aldehyde reductase (AFAR). Turkey liver cytosol also had quinone oxidoreductase (QOR) activity. Importantly, cytosol exhibited no measurable GST-mediated detoxification of microsomally activated AFB(1), indicating that turkeys are deficient in the most crucial AFB(1)-detoxification pathway. In total, our data indicate that the extreme sensitivity of turkeys to AFB(1) may be attributed to a combination of efficient AFB(1) activation and deficient detoxification by phase II enzymes, such as GSTs. PMID- 10814553 TI - A toxicokinetic study of inhaled ethylene glycol monomethyl ether (2-ME) and validation of a physiologically based pharmacokinetic model for the pregnant rat and human. AB - Exposures to sufficiently high doses of ethylene glycol monomethyl ether (2 methoxyethanol, 2-ME) have been found to produce developmental effects in rodents and nonhuman primates. The acetic acid metabolite of 2-ME, 2-methoxyacetic acid (2-MAA), is the likely toxicant, and, as such, an understanding of the kinetics of 2-MAA is important when assessing the potential risks to humans associated with 2-ME. A previously described physiologically based pharmacokinetic (PBPK) model of 2-ME/2-MAA kinetics for rats exposed via oral or iv administration was extended and validated to inhalation exposures. Pregnant Sprague-Dawley rats were exposed for 5 days (gestation days 11-15), 6 h/day, to 2-ME vapor at 10 and 50 ppm. Validation consisted of comparing model output to maternal blood and fetal 2 ME and 2-MAA concentrations during and following 5 days of exposure (gestation days 11-15). These concentrations correspond to a known no observed effect level (NOEL) and a lowest observed effect level (LOEL) for developmental effects in rats. The rat PBPK model for 2-ME/2-MAA was scaled to humans and the model (without the pregnancy component) was used to predict data collected by other investigators on the kinetics of 2-MAA excretion in urine following exposures to 2-ME in human volunteers. The partially validated human model (with the pregnancy component) was used to predict equivalent human exposure concentrations based on 2-MAA dose measures (maximum blood concentration, C(max), and average daily area under the 2-MAA blood concentration curve, AUC, during pregnancy) that correspond to the concentrations measured at the rat NOEL and LOEL exposure concentrations. Using traditional PBPK scale-up techniques, it was calculated that pregnant women exposed for 8 h/day, 5 days/week, for the duration of pregnancy would need to be exposed to 12 or 60 ppm 2-ME to produce maternal 2-MAA blood concentrations (C(max) or average daily AUC) equivalent to those in rats exposed to the NOEL (10 ppm) or LOEL (50 ppm), respectively. PMID- 10814554 TI - A toxicokinetic study of inhaled ethylene glycol ethyl ether acetate and validation of a physiologically based pharmacokinetic model for rat and human. AB - The solvents ethylene glycol monoethyl ether acetate (EGEEA) and ethylene glycol monoethyl ether (EGEE), at sufficiently high doses, are known to be rodent developmental toxicants, exerting their toxic effects through the action of their metabolite 2-ethoxyacetic acid (2-EAA). Thus risks associated with exposure to these compounds are best evaluated based on a measure of the internal dose of 2 EAA. The goals of the work reported here were to develop physiologically based pharmacokinetic (PBPK) models of EGEEA and EGEE for pregnant rats and humans. These models were used to identify human exposure levels (ppm in air) equivalent to the rat no observed effect level (NOEL) and lowest observed effect level (LOEL) for developmental effects (Hanley et al., 1984). We exposed pregnant Sprague-Dawley rats to concentrations of EGEEA corresponding to the NOEL and LOEL. Maternal blood, urine, and fetal tissue concentrations of EGEE and 2-EAA measured in these experiments were used to validate the rat EGEEA and EGEE models. Data collected by other researchers were used to validate the capabilities of the rodent EGEEA and EGEE models to predict the kinetics in humans. The models for estimating circulating blood concentrations of 2-EAA were considered valid based on the ability of the model to accurately predict 2-EAA concentrations in rat blood, urine, and fetal tissue. The human inhaled concentration equivalent to the rat NOEL for EGEEA (50 ppm) was predicted to be 25 ppm using the maternal blood average daily area under the curve (AUC) and 40 ppm using the maximum concentration achieved in maternal blood (C(max)). The human inhaled concentration equivalent to the rat LOEL for EGEEA (100 ppm) was determined to be 55 ppm using the maternal blood average daily AUC and 80 ppm using the maternal blood C(max). PMID- 10814555 TI - DMPS-arsenic challenge test. II. Modulation of arsenic species, including monomethylarsonous acid (MMA(III)), excreted in human urine. AB - The administration of sodium 2,3-dimercapto-1-propane sulfonate (DMPS) to humans chronically exposed to inorganic arsenic in their drinking water resulted in the increased urinary excretion of arsenic, the appearance and identification of monomethylarsonous acid (MMA(III)) in their urine, and a large decrease in the concentration and percentage of urinary dimethylarsinic acid (DMA). This is the first time that MMA(III) has been detected in the urine. In vitro biochemical experiments were then designed and performed to understand the urinary appearance of MMA(III) and decrease of DMA. The DMPS-MMA(III) complex was not active as a substrate for the MMA(III) methyltransferase. The experimental results support the hypothesis that DMPS competes with endogenous ligands for MMA(III), forming a DMPS-MMA complex that is readily excreted in the urine and points out the need for studying the biochemical toxicology of MMA(III). It should be emphasized that MMA(III) was excreted in the urine only after DMPS administration. The results of these studies raise many questions about the potential central role of MMA(III) in the toxicity of inorganic arsenic and to the potential involvement of MMA(III) in the little-understood etiology of hyperkeratosis, hyperpigmentation, and cancer that can result from chronic inorganic arsenic exposure. PMID- 10814556 TI - Enhanced allergic sensitization by residual oil fly ash particles is mediated by soluble metal constituents. AB - Epidemiological studies have demonstrated an association between elevated levels of particulate matter (PM) air pollutants and exacerbation of asthma symptoms. We have shown in a Brown Norway (BN) rat model of house dust mite (HDM) allergy that preexposure to residual oil fly ash (ROFA) particles enhanced the sensitization phase such that the secondary immune response and associated lung injury were increased after allergen challenge. To determine whether the metals present in ROFA mediated this effect, BN rats were intratracheally instilled with either ROFA (1000 microg) or acidified saline + NiSO(4) (105.12 microg), VSO(4) (98.2 microg), FeSO(4) (58.49 microg), or a mixture (Mix) of each metal. HDM-specific IgE was higher in the serum of the ROFA, Ni, V, and Mix groups than in the HDM group after challenge, and antigen-induced bronchoconstriction responses were increased in the Ni group. Lymphocyte proliferation to antigen was increased in the ROFA, Ni, and V groups compared to controls. Total protein and eosinophil peroxidase levels were elevated in the Fe group, and eosinophil numbers in the bronchoalveolar lavage fluid (BALF) were increased in the ROFA and Fe groups compared to HDM control. IL-5 and IL-13 mRNA expression was also increased in the lung tissue of all metal and ROFA-treated groups, while BALF IL-10 was elevated in the Fe and Mix groups, and IL-6 and TNF-alpha were elevated in the metal and ROFA-treated groups compared to controls. These results suggest that ROFA's metallic constituents mediate enhancement of sensitization to HDM and that pulmonary inflammation may play a role in this adjuvant effect. PMID- 10814557 TI - Nivalenol inhibits total and antigen-specific IgE production in mice. AB - Nivalenol (NIV) has been reported to induce hyperproduction of IgA, which is regulated by T-helper 2 cells (Th2); however, whether IgE production, which is under the regulation of Th2 cells, is induced by this compound remains largely unknown. We examined the effect of NIV on antigen-specific IgE production using ovalbumin (OVA)-specific T cell receptor alphabeta-transgenic mice. The mice produced significant amounts of total and antigen-specific IgE, IgG1, and IgA in serum when given OVA orally. Administration of NIV with OVA suppressed total IgE and OVA-specific IgE, IgG1, and IgA production significantly. Cytokine assay using splenocytes obtained from mice given the OVA plus NIV diet revealed that interleukin 4 (IL-4) production was suppressed and interleuin-2 (IL-2) production was enhanced. These results suggest that the inhibition of IL-4 production and enhancement of IL-2 production induced by NIV suppressed total and antigen specific IgE production. PMID- 10814558 TI - Stressful manipulations that elevate corticosterone reduce blood-brain barrier permeability to pyridostigmine in the Rat. AB - Pyridostigmine bromide (PB), a reversible inhibitor of acetylcholinesterase (AChE), is used for the treatment of myasthenia gravis. PB has also been provided to military personnel for preexposure protection against potential soman release. The entry of PB into the brain is typically minimal, but recently published data in mice suggest that a brief forced swim stress increases the permeability of the blood-brain barrier to PB. From these results, PB administered under stressful conditions was proposed to induce long-lasting central cholinergic deficits, potentially explaining the neurological and neuropsychological symptoms presented by some Gulf War veterans. In undertaking to replicate these results in the Long Evans rat, no evidence of a stress-potentiated central effect of PB, administered at doses up 5.0 mg/kg ip, was found. Three stress protocols were used: restraint, forced swim, or a combined restraint/forced swim. Wistar rats were also tested in some of the protocols to ensure that the results were generalizable across rat strains, and plasma corticosterone levels were measured to test the effectiveness of the stressors employed. In contrast to the previously reported findings in the mouse, stress significantly reduced the entry of PB into rat brain, as measured by reduced inhibition of AChE activity: a 12.5% reduction in whole brain AChE activity after treatment with 5.0 mg/kg PB under control conditions declined to 9% after stress exposure. It is apparent, therefore, that the interaction between stress and PB requires further study, and previous data should be reassessed before they are used as a basis for interpreting symptoms presented by veterans. PMID- 10814559 TI - Systematic reviews of chemotherapy and endocrine therapy in metastatic breast cancer. AB - Metastatic breast cancer is incurable but often responsive to treatment. There is little evidence-based consenus on when to use which treatments, in what combination and for how long. Systematic reviews were performed on 12 prospectively defined, clinically relevant research questions to support the development of evidence-based clinical practice guidelines. A comprehensive search of Medline from 1966 to 1996 identified over 1800 controlled trials. Eligibility and data extraction were performed independently by two blinded reviewers. Trial results were summarised by ratios of median survivals (RMS) and P -values for survival curve comparisons with meta-analysis by weighted combination of these statistics. Sixty-five publications reporting 97 treatment comparisons were included. There was moderate evidence that more rather than fewer cycles of chemotherapy improved survival (RMS:1.23, P -0.01). The evidence did not support: higher rather than lower doses of chemotherapy (or of endocrine therapy); any one class of endocrine agent over all others; multiple endocrine agents over a single agent; or, combined chemotherapy and endocrine therapy over either single modality. Only six trials assessed quality of life revealing better quality of life with more rather than fewer cycles of chemotherapy and with standard rather than lower doses of chemotherapy. These systematic reviews reveal counterintuitive evidence useful to everyday practice, in particular that more rather than fewer cycles of chemotherapy lead to better quality of life and longer survival. PMID- 10814560 TI - Immunomodulation therapy in colorectal carcinoma. AB - There has been much progress in the understanding of the relationship between the immune system and colorectal cancer. This has led to the use of immunomodulatory therapy in the adjuvant and palliative treatment of the condition. Although attempts at the use of non-specific immunomodulation with agents such as levamisole, cimetidine, alpha interferon and Bacillus Calmette-Guerin (BCG) have not produced significant clinical benefits when tested in randomized trials in both the adjuvant setting and for metastatic disease, promising results are being obtained with more specific therapy. Edrecolomab [corrected], a murine monoclonal antibody targeting the 17-1A antigen on malignant colorectal cells has produced a reduction in relapse and mortality rates when used as adjuvant treatment following surgery for Dukes' C colon cancer. Active specific therapy with autologous tumour vaccine administered with BCG has produced similar benefits in Dukes' B cancer. Both 3H1 anti-idiotypic antibody against carcinoembryonic antigen and 105AD7 antibody to gp72 glycoprotein have demonstrated in-vitro and in-vivo immune activation against tumour. Non-randomized studies postulate prolongation of survival using these antibodies in advanced disease. These agents are all currently being tested in randomized studies powered to detect meaningful survival differences and clinical benefit. Immune therapy offers the potential of low toxicity therapy in colorectal cancer and may have a role as an adjunct to conventional chemotherapy. PMID- 10814561 TI - Tumour vasculature as a target for anticancer therapy. AB - The development of a blood supply is crucial to the growth and metastasis of cancer. The factors involved in this are complex, however tumour hypoxia and macrophage infiltration are responsible for the synthesis of pro-angiogenic cytokines such as vascular endothelial growth factor (VEGF) and the fibroblast growth factors. These factors stimulate proliferation of vascular endothelial cells, the synthesis of proteases such as urokinase type plasminogen activator (uPA) and the matrix metalloproteases, which result in digestion of the extracellular matrix and allow endothelial cell invasion. Endothelial cell motility is promoted by binding of extracellular matrix proteins such as vitronectin and fibronectin to integrins expressed on the plasma membrane of endothelial cells. Interfering with any of these steps may inhibit the process of angiogenesis and drugs aimed at modulation of angiogenesis are currently undergoing evaluation in early clinical studies. This paper reviews our current understanding of angiogenesis and how it may be used as a target for the treatment of cancer. PMID- 10814562 TI - Selective oestrogen receptor modifiers (SERMs) and breast cancer therapy. AB - Antioestrogen therapy is currently receiving renewed interest for several reasons. Tamoxifen was introduced in the treatment of metastatic breast cancer more than three decades ago. The drug significantly reduces long term mortality and also reduces the risk of contralateral tumours when administered in early breast cancer. Five years of tamoxifen is now standard in adjuvant endocrine therapy, and the drug is currently being evaluated for breast cancer prevention. Despite this, several aspects regarding the pharmacology of the drug are still unclear, and the scientific rationale for dose selection has recently been challenged. Several novel antioestrogen compounds, called selective oestrogen receptor modifiers (SERMs), express selective oestrogen agonistic or antagonistic properties depending on the organ or test system evaluated. Some of these drugs, like raloxifene, do not seem to promote the development of endometrial cancer, although they still have selected oestrogen-like beneficial effects. This paper reviews the pharmacologic and the pharmacokinetic aspects of the different SERMs with particular emphasis on their potential use in therapy and prevention of breast cancer. PMID- 10814563 TI - Articles to appear in forthcoming issues of cancer treatment reviews PMID- 10814564 TI - Foamy viruses: between retroviruses and pararetroviruses. PMID- 10814565 TI - Interactions of HIV-1 nef with the mu subunits of adaptor protein complexes 1, 2, and 3: role of the dileucine-based sorting motif. AB - HIV-1 Nef interacts with cellular adaptor protein (AP) complexes and their medium (mu) subunits. However, the role of the dileucine-based sorting motif within Nef in these interactions has been incompletely characterized. Here, yeast two-hybrid assays indicated that HIV-1 Nef interacted not only with the mu subunits of AP-1 and AP-2, but also with that of AP-3. The interactions with mu1 and mu3 were markedly stronger than the interaction with mu2. Leucine residues of the sorting motif were required for the interactions with mu3 and mu2 and contributed to the interaction with mu1. Confocal immunofluorescence microscopy indicated that Nef, AP-1, and AP-3 (but not AP-2) were concentrated in a juxtanuclear region near the cell center, potentially facilitating interaction between Nef and the mu1 and mu3 subunits. However, leucine residues of the sorting motif were not required for this subcellular localization of Nef. These data suggest that the dileucine motif, required for optimal viral replication, functions through interactions with a variety of AP complexes, including AP-3, potentially by recruiting adaptor complexes to subcellular locations specified by additional determinants in the Nef protein. PMID- 10814566 TI - Phytoreovirus T = 1 core plays critical roles in organizing the outer capsid of T = 13 quasi-equivalence. AB - The structures of the double-shelled rice dwarf virus and of its single-shell core have been determined by cryoelectron microscopy and image reconstruction. The core carries a prominent density located at each of the icosahedral faces of its T = 1 lattice. These protrusions are formed by outer shell trimers, tightly inserted at the threefold positions of the core. Such configuration of the core may guide the assembly of the outer shell, aided by lateral interactions between its subunits, into a T = 13 lattice. The organization of the phytoreovirus capsid elucidates for the first time a general model for assembling two unique T numbers of quasi-equivalence. PMID- 10814567 TI - The vaccinia virus A36R protein is a type Ib membrane protein present on intracellular but not extracellular enveloped virus particles. AB - Vaccinia virus gene A36R encodes a 45-kDa protein that is conserved in orthopoxviruses. A virus lacking the A36R protein formed a small plaque, was unable to induce the polymerization of actin tails, and was avirulent in vivo. Here we present a further characterization of the A36R protein by in vitro transcription and translation and analysis of infected cells by confocal microscopy and immunoelectron microscopy of cryosections using a monoclonal antibody raised against the C-terminal domain of the A36R protein. Translation of the A36R mRNA in vitro produced a protein of the same size whether or not the translation reaction was performed in the presence of canine pancreatic microsomes. However, the polypeptide synthesized in the presence of microsomes was associated integrally with the membrane and was sensitive to digestion by exogenous protease without permeabilization of the membrane with detergent, indicating that the majority of the protein is exposed on the outside of the vesicle. Consistent with this, immunofluorescent analysis of virus-infected cells demonstrated that the C-terminal domain of A36R was not exposed on the cell surface but was detected once the cell membrane was permeabilized. Immunoelectron microscopy of cryosections of infected cells showed that the protein was absent from IMV particles but present on intracellular enveloped virus (IEV) particles, predominantly on the cytosolic face of the IEV outer membrane. Where cell associated enveloped virus (CEV) particles were attached to the cell surface, the A36R protein was detected only on the cytosolic surface of the plasma membrane where the virus particle remained attached to the cell and not elsewhere on the plasma membrane or on the CEV particle. A36R and actin copurified with EEV particles due to the association of fragments of cellular membranes with the EEV particles. Therefore, A36R represents the first example of a virus-encoded protein that is present on IEV but not CEV particles. PMID- 10814568 TI - Sequencing of peach latent mosaic viroid variants from nine North American peach cultivars shows that this RNA folds into a complex secondary structure. AB - We sequenced 34 new peach latent mosaic viroid (PLMVd) variants isolated from nine different peach cultivars. This study provides the widest view of PLMVd diversity reported to date and includes the original characterization of North American variants, which cannot be differentiated from European sequences. PLMVd appears as a species in which each isolate is a complex mixture of RNAs. Analysis of base-pair covariations supports the hypothesis that PLMVd folds into a complex branched structure with the potential of including three new pseudoknots. The resulting "globular-like" structure is in contrast to the rod-like one adopted by most other viroids. PMID- 10814569 TI - Specific interaction between the hepatitis delta virus RNA and glyceraldehyde 3 phosphate dehydrogenase: an enhancement on ribozyme catalysis. AB - Replication of hepatitis delta virus (HDV) RNA occurs in the nuclei of infected cells. The replication is mediated by cellular factors containing an RNA polymerase II-like enzyme activity through a double rolling-circle mechanism and is regulated by delta antigens. In this study, UV cross-linking experiments were carried out to examine interactions between HDV RNA and proteins present in HeLa nuclear extract. Cellular proteins with molecular mass of 23 (p23), 36 (p36), 38 (p38), and 58 (p58) kDa bound to full-length HDV RNA of both genomic and antigenomic strands. Deletion analysis on the antigenomic strand mapped the interacting domain within a 79-nucleotide fragment but not at the ends of the rod shaped viral RNA structure. The specificity of the RNA-protein interactions was demonstrated by competition experiments and the specific HDV RNA-binding proteins were purified through column chromatography. Electrophoresis mobility shift assay with the purified fractions demonstrated that the interaction between p36 and HDV RNA was relatively stable even in the presence of 0.5 M NaCl. Biochemical analysis including protein microsequencing identified the p36 as glyceraldehyde 3 phosphate dehydrogenase (GAPDH). RNase footprinting indicated that the UC-rich domain between nucleotides 379 and 414 of the HDV antigenomic RNA was involved in the GAPDH binding. Functional studies further demonstrated an enhancing effect of GAPDH on the ribozyme activity of HDV antigenomic RNA. In addition, in the presence of HDV RNA cellular GAPDH relocalized from the cytoplasm to the nucleus where HDV replication occurs. These results suggest that GAPDH is involved in the replication of HDV. PMID- 10814570 TI - Formation of surface tubules and fluorescent foci in Arabidopsis thaliana protoplasts expressing a fusion between the green fluorescent protein and the cauliflower mosaic virus movement protein. AB - The movement protein (MP) of cauliflower mosaic virus (CaMV) is a multifunctional protein that potentiates the cell-to-cell and long distance movement of the virus. Functional domains in the CaMV MP were determined by analyzing deletions in green fluorescent protein (GFP)-MP fusions transfected into Arabidopsis thaliana leaf protoplasts. GFP-MP accumulated at fluorescent foci at the cell periphery and in tubular structures extending from the protoplast surface. A region located near the center of MP was required for targeting GFP-MP to foci, whereas a larger region extending nearly to the N-terminus was needed for tubule formation. Cytoskeletal assembly inhibitors did not disrupt tubule formation or the accumulation of GFP-MP at foci, but brefeldin A, which disrupts the plant cell endomembrane system, did interfere with the formation of tubules but not foci. PMID- 10814571 TI - Abnormalities of cornified cell envelopes isolated from human papillomavirus type 11-infected genital epithelium. AB - Keratinocytes are the predominant cells in human skin. As keratinocytes differentiate, the nuclei are lost and the cornified cell envelope (CCE) develops, forming a covalently cross-linked, insoluble structure under the cell membrane. Layers of anuclear CCEs in the stratum corneum provide a barrier against water loss and mechanical damage and are a first line of immunologic defense. Infection of keratinocytes with human papillomaviruses (HPVs) induces proliferation and abnormalities including retention of nuclei in the stratum corneum and perinuclear halo formation. For effective transmission, HPV virions must be released from the CCE, a normally very durable structure. Therefore, it is likely that HPV infection affects the CCE in a manner that would facilitate virion release. To investigate the effects of HPV 11 infection on morphology and fragility, CCEs were purified from infected and uninfected epithelium. CCEs isolated from uninfected epithelium were smooth, cuboidal, and sonicated into long coiled structures. In contrast, CCEs from HPV 11-infected epithelium were irregular in size and shape, with rough edges, and sonicated into small fragments. In addition, the thickness of CCEs from HPV 11-infected tissue was 65% that of uninfected epithelium. Immunohistochemical analysis demonstrated that in contrast to uninfected epithelium, loricrin, the major component of the CCE, was abnormally distributed in the differentiated layers of HPV 11-infected epithelium. We conclude that in addition to the previously described epithelial abnormalities induced by HPV, the CCE is also affected by infection in ways that may facilitate transmission of virus from person to person. PMID- 10814572 TI - Temperature dependence of fusion by sendai virus. AB - Studies of the temperature dependence of liposome fusion by Sendai virus indicate that fusion occurs maximally at 55 degrees C. The fusion capacity of the virus is also inactivated maximally by preincubation at this temperature and, under the same conditions, the F glycoprotein becomes resistant to proteolysis. By analogy with the activation at elevated temperatures of fusion by influenza virus our results suggest that temperature is also a variable in the activation of fusion by paramyxoviruses and possibly in the activation of other members of the group of viruses that includes myxo-, paramyxo-, retro-, and filoviruses, which all contain cleaved, trimeric fusion glycoproteins. PMID- 10814573 TI - Incomplete HIV-1 activation in latently infected U1 cells demonstrated by double in situ hybridization. AB - Triple combination antiretroviral therapy can reduce HIV-1 infection to a relatively small pool of latently infected cells. To eliminate this residual source of virus, new therapies designed to activate latently infected cells are currently being tested. We therefore investigated the kinetics of in vitro HIV-1 RNA induction using chronically infected U1 cells. A new two-probe fluorescence in situ hybridization (double ISH) method was devised to simultaneously assess total HIV-1 RNA (T-RNA) and unspliced HIV-1 RNA (U-RNA) expression in individual cells. Activation of the U1 cells resulted in increasing expression of T-RNA between 0 and 24 h with lagging expression of U-RNA between 6 and 30 h. Both the positive area per cell and the number of positive cells increased with time. Although activation induced 98.5% of the cells to express HIV-1 T-RNA by 24 h, 52% remained negative for U-RNA. In contrast, 100% of 8E5 cells, which constitutively express HIV-1, scored positive for U-RNA as well as T-RNA with the double ISH. This study provides, for the first time, a semiquantitative cell-by cell analysis of HIV-1 mRNA subsets in latently infected cells. Our results establish the advantages of using double fluorescence ISH to study gene expression and demonstrate that chronically infected U1 cells remain in a partially induced state despite potent activation. PMID- 10814574 TI - Functions of the 126- and 183-kDa proteins of tobacco mosaic virus. AB - Tobacco mosaic virus produces two proteins that contain domains similar to the methyltransferase (MT) and helicase (HEL)-like domains of the replicase associated proteins of other RNA viruses. The more abundant 126-kDa protein contains only the MT and HEL-like domains, whereas the 183-kDa readthrough protein additionally contains the polymerase domain. We examined the functions of these proteins by constructing a bipartite system to express the 126- and 183-kDa proteins from separate RNAs. Mutants expressing the 183-kDa protein recognized promoters for negative- and positive-stranded RNA synthesis, transcribed subgenomic mRNAs, capped RNAs, synthesized proteins, moved cell to cell within the plant, and replicated defective RNAs (dRNAs). The principal function of the 126-kDa protein was to increase the rate of replication approximately tenfold. The 126-kDa protein appeared to function primarily in cis, and production of the 126-kDa protein in trans did not enhance replication of the helper virus. dRNAs producing a functional 126-kDa protein were replicated efficiently by helper viruses that produced only the 183-kDa protein but not by wild-type virus, suggesting that efficient replication required the 183-kDa protein to form a heterodimer with the 126-kDa protein already bound to the target dRNA. PMID- 10814575 TI - The coxsackie-adenovirus receptor (CAR) is used by reference strains and clinical isolates representing all six serotypes of coxsackievirus group B and by swine vesicular disease virus. AB - Group B coxsackieviruses are etiologically linked to many human diseases, and cell surface receptors are postulated to play an important role in mediating their pathogenesis. The coxsackievirus adenovirus receptor (CAR) has been shown to function as a receptor for selected strains of coxsackievirus group B (CVB) serotypes 3, 4, and 5 and is postulated to serve as a receptor for all six serotypes. In this study, we demonstrate that CAR can serve as a receptor for laboratory reference strains and clinical isolates of all six CVB serotypes. Infection of CHO cells expressing human CAR results in a 1000-fold increase in CVB progeny virus titer compared to mock transfected cells. CAR was shown to be a functional receptor for swine vesicular disease virus (SVDV), as CHO-CAR cells but not CHO mock transfected controls were susceptible to SVDV infection, produced progeny SVDV, and developed cytopathic effects. Moreover, SVDV infection could be specifically blocked by monoclonal antibody to CAR (RmcB). SVDV infection of HeLa cells was also inhibited by an anti-CD55 MAb, suggesting that this virus, like some CVB, may interact with CD55 (decay accelerating factor) in addition to CAR. Finally, pretreatment of CVB or SVDV with soluble CAR effectively blocks virus infection of HeLa cell monolayers. PMID- 10814576 TI - Identification and molecular characterization of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus genomic region encoding the regulatory genes pkip, p47, lef-12, and gta. AB - Choristoneura fumiferana multicapsid nucleopolyhedrovirus (CfMNPV) is a baculovirus pathogenic to spruce budworm, the most damaging insect pest in Canadian forestry. CfMNPV is less virulent to its host insect and its replication cycle is slower than the baculovirus type species Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) but the basis of these characteristics is not known. We have now identified, localized, and determined the sequence of the region of CfMNPV carrying potentially important regulatory genes including p47, lef-12, gta, and pkip. DNA database searches revealed that this region of CfMNPV is most closely related to the homologous OpMNPV genes. Transcription analysis demonstrated that CfMNPV P47 is encoded by a 1.6-kb transcript, LEF-12 is encoded by a 2.6-kb transcript, and GTA is encoded by a 2.1-kb transcript. Transcripts for these genes were detectable at 6 h postinfection but all of them showed a burst in expression levels between 12 and 24 h postinfection corresponding to the time of initiation of CfMNPV DNA replication. A polyclonal antibody, raised against CfMNPV P47, detected a nuclear 43-kDa polypeptide from 12 to 72 h postinfection, demonstrating that the CfMNPV p47 gene product is first expressed at a time corresponding to the burst of transcriptional activity between the early and the late phases. Both AcMNPV and CfMNPV P47 translocate to the nucleus of infected cells. PMID- 10814577 TI - Electron microscopy of the human respiratory syncytial virus fusion protein and complexes that it forms with monoclonal antibodies. AB - Full-length fusion (F) glycoprotein of human respiratory syncytial virus (HRSV) and a truncated anchorless mutant lacking the C-terminal 50 amino acids were expressed from vaccinia recombinants and purified by immunoaffinity chromatography and sucrose gradient centrifugation. Electron microscopy of full length F protein in the absence of detergents revealed micelles, (i.e., rosettes) containing two distinct types of protein rods, one cone-shaped and the other lollipop-shaped. Analysis of membrane anchorless F molecules indicated that they were similar to the cone-shaped rods and that rosettes, which they formed on storage, were made up of lollipop-shaped rods. The two forms of F protein may represent different structures that the molecule may adopt before and after activation for its role in membrane fusion. Studies of complexes of these structures with monoclonal antibodies of known specificity provide information on the three-dimensional organization of antigenic sites on the F protein and confirm the oligomeric structure, possibly trimeric, of both full-length F and membrane anchorless F. PMID- 10814578 TI - Characterization of a membrane-associated protein implicated in visna virus binding and infection. AB - The identity of the cellular receptor(s) for visna virus, an ovine lentivirus, is currently unknown; however, previous studies from our laboratory have identified membrane-associated proteins expressed selectively in susceptible cells which bind visna virus. Moreover, a polyclonal antibody (2-23), raised against a 45-kDa visna virus binding protein, bound specifically to the surface of susceptible cells in immunofluorescence assays and significantly reduced binding of visna virus to cells (S. E. Crane et al., 1991, J. Virol., 65, 6137-6143). In this report we extend our studies of this antibody (2-23), showing both that 2-23 significantly reduces visna virus infection of susceptible cells and that 2-23 immunoprecipitates a putative protein complex consisting of a prominent 30-kDa protein, as well as the 45-kDa immunogen, specifically from radiolabeled virus susceptible sheep cells. Further, we demonstrate that the 30-kDa protein is a membrane-associated proteoglycan substituted with a chondroitin sulfate glycosaminoglycan (GAG) chain(s) and that treatment of susceptible cells with an inhibitor of GAG synthesis significantly reduces visna virus production. Collectively, these data support a role for a proteoglycan in visna virus cell binding and infection. PMID- 10814579 TI - Isolation, cloning, and complete nucleotide sequence of a phenotypically distinct Brazilian isolate of human T-lymphotropic virus type II (HTLV-II). AB - Analysis of human T-lymphotropic virus type II (HTLV-II) isolates from North America and Europe have demonstrated the existence of two molecular subtypes of the virus, HTLV-IIa and HTLV-IIb. Recently, studies on HTLV-II infections in Brazil have revealed isolates that are related phylogenetically to the HTLV-IIa subtype but have a HTLV-IIb phenotype with respect to the transactivating protein, tax. To more clearly define this relationship, HTLV-II was isolated from peripheral blood of an IVDA from Sao Paulo, Brazil (SP-WV), and the complete provirus was cloned and sequenced. Comparison of HTLV-II(SP-WV) nucleotide sequences to other available complete HTLV-II proviral sequences revealed that HTLV-II(SP-WV) is most closely related to HTLV-II(Mo), the prototypic HTLV-IIa subtype sequence. Phylogenetic analysis of LTR, env, and tax regions unequivocally demonstrated that HTLV-II(SP-WV) and all other Brazilian sequences examined are members of the IIa subtype. The predicted amino acid sequences of the major coding regions of HTLV-II(SP-WV) are also most closely related to HTLV II(Mo), with the important exception of tax. The tax protein encoded by HTLV II(SP-WV) is 96-99% identical to the tax of IIb isolates and is similar in that it has an additional 25 amino acids at the carboxy-terminus compared to the HTLV II(Mo) tax with which it shares 91% identity. Analysis of tax stop codon usage of a number of HTLV-IIa isolates from North American, Europe, and Brazil demonstrated that isolates from the last region appear to be unique in their extended tax phenotype. It could be demonstrated that the extended tax proteins in the HTLV-IIb and Brazilian isolates had equivalent ability to transactivate the viral LTR, and studies with deletion mutants indicated that the extended C terminus is not essential for transactivation. In contrast, the HTLV-IIa tax was found to have a greatly diminished ability to transactivate the viral LTR, which appeared to be a consequence of reduced expression of the protein. The studies show that although the Brazilian strains do not represent an entirely new subtype based on nucleotide sequence analysis they are a phenotypically unique molecular variant within the HTLV-IIa subtype. PMID- 10814580 TI - Decreased vaginal disease in J-chain-deficient mice following herpes simplex type 2 genital infection. AB - J-chain-deficient (Jch(-/-)) mice were used to study the role of polymeric IgA (pIgA) in primary disease and protective immunity following genital herpes simplex type 2 (HSV-2) infection. Vaginal IgA in the Jch(-/-) mice was composed primarily of monomeric IgA and was not associated with secretory component (SC). In contrast, vaginal IgA in wild-type (WT) mice was predominantly polymeric and bound to SC. Following HSV-2 genital infection, the Jch(-/-) mice consistently exhibited fewer vaginal symptoms (P = 0.010) and mortality (P = 0.075) than did the WT mice. The variation in disease expression could not be explained by differences in local viral replication, since titers in vaginal wash fluid were comparable. To assess the effect of J chain deficiency on protective immunity, WT and Jch(-/-) mice were immunized intravaginally with attenuated HSV-2, challenged intravaginally with wild-type virus 5 weeks later, and evaluated for vaginal infection and neurological disease. Although the Jch(-/-) mice had reduced vaginal HSV-specific IgA and IgG levels following immunization, both WT and Jch( /-) mice were protected from symptoms following wild-type virus challenge. We conclude that pIgA is not required for protective immunity against genital HSV-2 disease and that J chain deficiency offers some protection against symptoms following primary HSV-2 genital infection. PMID- 10814581 TI - Competitive binding to a charged leucine motif represses transformation by a papillomavirus E6 oncoprotein. AB - E6 oncoproteins from HPV-16 and bovine papillomavirus type 1 (BPV-1) bind to similar leucine-rich peptides termed charged leucine motifs found on the cellular focal adhesion protein paxillin and the E3 ubiquitin ligase E6AP. BPV-1 E6 (BE6) mutants that do not bind to paxillin are defective at inducing cellular transformation. It is possible, however, that BE6 mutants that do not bind paxillin are defective for transformation for an unrelated reason than the ability to bind to charged leucine motifs. To address the role of BE6 interaction with charged leucine motifs, we fused a BE6-binding charged leucine motif to the amino terminus of BE6, thereby creating an autoinhibitory binding domain. We found that the fusion protein failed to bind to paxillin or transform murine C127 cells. Mutation of the amino terminal binding motif in the fusion protein restored both interaction with paxillin and transformation. This demonstrates that BE6 transformation requires binding to charged leucine motifs on particular cellular proteins and that transformation by papillomavirus oncoproteins can be repressed by competitive interactions with charged leucine motifs. PMID- 10814582 TI - Re-emergence of vesicular stomatitis in the western United States is associated with distinct viral genetic lineages. AB - Phylogenetic analysis of partial phosphoprotein and glycoprotein gene sequences showed that a single genetic lineage of vesicular stomatitis virus (VSV) serotype New Jersey (NJ) caused the 1995 and 1997 outbreaks of vesicular stomatitis (VS) in the western United States. While distinct from VSV-NJ strains causing previous outbreaks in the western United States and those circulating in feral swine in the southeastern United States, this lineage was closely related to viral lineages circulating in the Mexican states of Guerrero, Veracruz, and Oaxaca in 1996, 1989, and 1984 respectively. In 1997 and 1998, VSV serotype Indiana 1 (IN1) re-emerged in the western United States after 30 years. Viruses causing these outbreaks grouped within a single genetic lineage distinct from VSV-IN1 isolates causing outbreaks in the western United States in 1929 and 1956 but closely related to a strain circulating in the state of Colima in central Mexico in 1997. Our data showed that sporadic VS outbreaks in the western United States are caused by genetically distinct viral lineages closer to those circulating in enzootic areas of central and southern Mexico than to those causing previous outbreaks in the United States. The genetic evidence and temporal distribution of outbreaks are not consistent with a pattern of long-term maintenance of VSV in the western United States. PMID- 10814583 TI - Characterization of murine coronavirus neutralization epitopes with phage displayed peptides. AB - Phage-displayed peptide libraries were used to map immunologically relevant epitopes on the surface (S) glycoprotein of a neurotropic murine coronavirus (MHV A59). Three in vitro virus-neutralizing and in vivo protective mAbs against either continuous or discontinuous epitopes on the S glycoprotein were used to screen 12 different peptide libraries expressed on the pVIII major coat protein of the fd filamentous bacteriophage. Consensus sequences that matched short sequences within the S glycoprotein were identified. The sequence of a tight binding, mAb-selected peptide suggested the location of a discontinuous epitope within the N-terminal S1 subunit. Several tightly binding phage were amplified and used directly as immunogens in BALB/c and C57BL/6 mice. Partial protection of C57BL/6 mice against a lethal acute virus infection was achieved with a phage preparation that displayed a linear epitope. Protection correlated with the presence of sufficient levels of specific antiviral antibodies recognizing the same immunodominant domain and 13-mer peptide, located within the C-terminal S2 subunit, as the selecting mAb. Thus, the direct use of phage-displayed peptides to evaluate protective antiviral immune responses complements their use to characterize antibody-binding epitopes. This is the first evaluation of protective immunization induced by mAb-selected phage-displayed peptides. PMID- 10814585 TI - A heterogeneous population of defective RNAs is associated with lettuce infectious yellows virus. AB - Preparations of dsRNAs and virion RNAs extracted from Nicotiana clevelandii plants infected with the bipartite Lettuce infectious yellows virus (LIYV) were found to contain multiple LIYV RNA species. In addition to the two LIYV genomic RNAs, three types of RNAs were observed: (a) 3' coterminal subgenomic RNAs; (b) RNAs containing LIYV RNA 1 or RNA 2 5' terminus but lacking the 3' terminus; and (c) RNAs with both LIYV RNA 2 3' and 5' termini but each with a central extensive deletion, a structure typical of defective RNAs (D RNAs). No D RNA-like RNAs were detected for LIYV RNA 1. A reverse transcription followed by polymerase chain reaction (RT-PCR) strategy was used to clone from virion RNAs several LIYV RNA 2 D RNAs as cDNAs. Nucleotide sequence analysis of 43 cloned cDNAs showed in some D RNAs the presence of a stretch of 1-5 nt in the junction site that is repeated in the genomic RNA 2 in the two positions flanking the junction site or in close proximity. Some D RNAs contained in the junction site one or several extra nucleotides not present in the LIYV genomic RNA 2. Two of the cloned cDNAs were used to generate in vitro transcripts, and infectivity studies showed that both D RNAs were replication competent in protoplasts when coinoculated with LIYV RNAs 1 and 2 or with only LIYV RNA1. Neither D RNA showed obvious effects upon LIYV RNA 1 and RNA 2 accumulation in coinfected protoplasts. These data suggest that LIYV infections contain a heterogeneous population of LIYV RNA 2 D RNAs, and some are encapsidated into virions. PMID- 10814584 TI - Hepatitis C virus core protein promotes immortalization of primary human hepatocytes. AB - Hepatitis C virus (HCV) core protein has many intriguing properties as a viral factor and is implicated in cell growth regulation. In this study, the cell growth regulation potential of HCV core protein was investigated by introduction of the core genomic region into primary human hepatocytes, a natural host for virus replication and tropism. Core-transfected primary human hepatocytes displayed altered cell morphology resembling that of low-differentiated epithelial cells. Those cells retained an immortalized phenotype and exhibited continuous growth after more than 50 passages over 2 years. Stable hepatocyte transfectants exhibited albumin secretion and HCV core protein expression. Telomerase activity, a characteristic of immortalized or transformed cells, was evident in the transfected hepatocytes immediately after senescence. Anchorage independent growth of the immortalized hepatocytes provided further evidence for a transformed phenotype. Results from these studies suggest that the HCV core protein promotes primary human hepatocytes to an immortalized phenotype, which may predispose cells over an extended period of time to undergo a transforming event. Thus, HCV core protein appears to contribute to virus-mediated pathogenesis in a persistently infected host. PMID- 10814586 TI - Hydrolytic action of alpha-amylase on high-amylose starch of low molecular mass. AB - High-amylose starches of low average degree of polymerization (d-p 61-71), formed as fine granules by interaction of Bacillus macerans cyclodextrin glucanotransferase with alpha-cyclodextrin (CD) at 2-70 degrees C, are highly insoluble in water and not gelatinizable under normal cooking conditions (100 degrees C). Samples of CD-derived starches, both cooked and uncooked, were subjected to hydrolysis in vitro by human salivary alpha-amylase at 37 degrees C under conditions chosen to resemble those in the human intestinal lumen. Released low-molecular-mass saccharides were determined quantitatively by HPLC and the results compared with those from similar studies with natural starches. Among uncooked starches, CD-derived starch showed very low reactivity towards alpha amylase, along with potato starch and a high-amylose hybrid corn starch(64% amylose). Cooking greatly enhanced reactivity of natural starches, but only moderately increased reactivity of CD-derived starches. Susceptibility to hydrolysis of cooked starches increased in the following general order: CD derived starch (approximately 100% amylose)<100%corn amylose (isolated by the butan-1-ol method)Thr. This change impeded the co-expression of a 48-kDa by-product from an internal translation site. Also, a second 58-kDa by product was identified as a GAD65 C-terminal proteolytic fragment that co purifies with thioredoxin-M161T GAD65. The new GAD65 variant was expressed and easily purified, yielding an antigen that performed equally or better than wild type GAD65 in the reference radiobinding assay for GADA. The procedure provides an inexpensive source of large amounts of fully active and immunochemically competent GAD65. PMID- 10814591 TI - Purification of native enolase from medically important Candida species. AB - The 48 kDa glycolytic enzyme, enolase, has been identified as an immunodominant antigen in Candida albicans infections. It has also been identified as an important fungal allergen. Enolase from a number of medically important Candida species has been purified using a two-step anion- and cation-exchange chromatography method that was preceded by an organic extraction. The enolases purified by this method have a high specific activity and the procedure is 40% efficient, with an average of 5 mg of enolase/g of Candida cells. The purification of native enolase from medically important Candida species will enable the immunological significance and interspecies relationships of this major fungal antigen to be investigated. PMID- 10814592 TI - Reverse transcription of an RNA genome from databasing paper (FTA(R)). AB - The instability of RNA in solutions during storage and travel is an impediment to its utilization in routine diagnostics. A robust and simple approach to the problem of RNA protection and processing is offered by storage of RNA desiccated with processing procedures that do not solublize the RNA until the beginning of reverse transcription. The feasibility of this general approach was tested with coxsackievirus B4 (CVB-4) from blood or culture fluid held on a storage and transport medium (FTA(R)) and analysed by reverse transcriptase PCR (RT-PCR) without removing the RNA from the FTA(R) until reverse transcription. Phase trapping techniques based on water-miscible solvents such as ethanol or phenol were compared with simple buffers and concentrated lithium chloride solutions. RT PCR detection of viral RNA reached a sensitivity of approximately 0.1 fg, which is comparable with other non-nested PCR techniques. Whole blood as a virus vehicle significantly interfered with CVB-4 detection, but to an acceptable degree. Desiccation-storage of the RNA of CVB-4 appears to be unaffected by weeks on the storage medium under ambient conditions. These characteristics indicate that this approach forms a credible developmental base for RNA-based pathogen diagnostics with particular application to the problem of transporting potentially infectious body fluids to a centralized laboratory for analysis. PMID- 10814593 TI - Expression of biologically active human butyrylcholinesterase in the cabbage looper (Trichoplusia ni). AB - This investigation examined the utility of three recombinant protein-expression systems (COS cells, insect cells and insect larvae) to cost-effectively produce biologically active human butyrylcholinesterase (BuChE). It was determined that baculovirus-infected insect cells (Sf9 and High 5) expressed 3.5- and 8.2-fold, respectively, more active enzyme than COS-7 cells. Baculovirus-infected cabbage looper (Trichoplusia ni) insect larvae produced over 26 times more than High 5 cells; in fact, one baculovirus-infected insect larva provided more active protein than 100 ml of insect cell culture. Analysis of the larvally expressed proteins revealed that the vast majority of BuChE expressed was inactive due to extensive degradation that occurred in vivo. However, the active form of BuChE does have enzyme kinetics similar to those of its human serum counterpart. Cabbage looper larvae were also examined for their ability to serve as an in vivo animal model to study protection against anti-cholinesterase toxicity. This was unsuccessful due to their high tolerance to the very toxic organophosphorus compounds tested. This tolerance may be attributed at least in part to a novel endogenous organophosphorus acid anhydride hydrolase activity that is capable of hydrolysing the chemical-warfare nerve agents sarin (isopropyl methylphosphonofluoridate) and soman (pinacolyl methylphosphonofluoridate). These results show that cabbage looper larvae can serve as an inexpensive recombinant protein-expression system for human BuChE. PMID- 10814594 TI - Introduction of thiol-reactive structures on to soluble and insoluble proteins. AB - When proteins containing disulphide groups were oxidized with magnesium monoperoxyphthalate at acidic pH, they acquired the property of binding thiol compounds. This was the case with the insoluble protein keratin, chosen for having a large number of disulphide bridges, and with soluble ones like BSA and immunoglobulins. The potential applications of some of these modified proteins for the preparation of soluble bioconjugates have been explored. As a particular example of an application, the immobilization of activated IgG on to solid phases might provide a new way for preparing immunoadsorbents. PMID- 10814595 TI - Purification and properties of a lipase from Cephaloleia presignis (Coleoptera, chrysomelidae). AB - A novel lipase from the insect Cephaloleia presignis was purified by a procedure involving ammonium sulphate precipitation, and Phenyl Toyopearl 650M, DEAE-5PW and hydrophobic-interaction column chromatographies. The purified lipase was homogeneous with a molecular mass of 31000 Da by SDS/PAGE and of 29000 Da by gel filtration on a Superose 12 column. The enzyme was identified as a glycoprotein with a pI of 6.9. The enzyme unspecifically liberated short-chain to long-chain fatty acids from p-nitrophenyl esters, methyl esters and triglycerides. The N terminal 28 amino acid residues were determined as AGTLGYATRHVLPIFTLDDYTGSNEMWG, which showed no similarity with known proteins, suggesting that the purified lipase may belong to a novel class of hydrolases. PMID- 10814596 TI - Scale-up process for expression and renaturation of recombinant human epidermal growth factor from Escherichia coli inclusion bodies. AB - A cDNA encoding mature epidermal growth factor (EGF) was isolated and cloned into a pQE30 vector in which the His(6)-tagged EGF was expressed. pH-stat feeding of concentrated medium at the time of isopropyl beta-D-thiogalactoside induction and slug-feedings of the enriched medium during the induction resulted in a higher cell density and specific expression. Using a simple refolding protocol that consisted of 1 mM L-cysteine addition for a 1-h reduction followed by 5 mM L cystine addition for oxidative refolding, we were able to convert nearly all EGF monomers into the oxidized form. Also, there folding aggregate was converted into the monomeric form. Approx. 50% overall yield was obtained from the dissolved inclusion bodies to a single peak under FPLC. We hope that the result of this study may provide information that is useful for the scale-up of the recombinant human EGF production process. PMID- 10814597 TI - Purification and characterization of biosurfactants from Nocardia sp. L-417. AB - The Nocardia sp. L-417 strain grown with n-hexadecane as a carbon source produced two types of biosurfactant that have different characteristics. These biosurfactants were purified by procedures that included ammonium sulphate fractionation, chilled acetone and hexane treatments, silica-gel column chromatography and Sephadex LH-20 gel filtration. The purified biosurfactants were very stable over a broad range of pHs (2-12) and temperatures (100 degrees C, 3 h). The biosurfactant type I had strong properties as an emulsifying agent and as an emulsion-stabilizing agent, whereas type II had a strong ability to reduce surface tension. PMID- 10814598 TI - Recent articles of interest to biotechnology in other PortlandPress journals PMID- 10814599 TI - Alveolar-capillary membrane dysfunction in chronic heart failure: pathophysiology and therapeutic implications. AB - Chronic heart failure (CHF) disturbs the alveolar-capillary interface and increases the resistance to gas transfer. Alveolar-capillary membrane conductance (D(M)) and capillary blood volume (V(c)) are subcomponents of the lung diffusion capacity. Elevation of the capillary pressure causes alveolar-capillary membrane stress failure (i.e. increase in capillary permeability to water and ions, and disruption of local regulatory mechanisms for gas exchange), leading to a decrease in D(M), an increase in V(c) and subsequent impairment of diffusion capacity. Renewed recent interest in abnormalities in lung diffusion in patients with CHF has brought about new pathophysiological insights. A significant contribution of the altered gas transfer to the pathogenesis of exercise limitation and ventilatory abnormalities has been reported, and D(M) has been identified as the best lung function predictor of oxygen uptake at peak exercise. This review examines the pathophysiological and clinical significance of assessing lung diffusion capacity in patients with CHF. PMID- 10814600 TI - Sumatriptan reduces exercise capacity in healthy males: a peripheral effect of 5 hydroxytryptamine agonism? AB - 5-Hydroxytryptamine (5-HT; serotonin) has been implicated in the perception of exercise-induced fatigue. Sumatriptan is a selective 5-HT(1B/D) receptor agonist which does not cross the blood-brain barrier. The aim of the present study was to determine the effect of sumatriptan on exercise capacity. Ten healthy male subjects (mean age 28.4+/-10.8 years) performed a maximal treadmill exercise test according to the Bruce protocol with expired gas analysis on two occasions. Either 6 mg of sumatriptan or placebo was administered subcutaneously in a randomized, double-blind, placebo-controlled, cross-over design. Exercise time was greater after placebo compared with sumatriptan [914 and 879 s respectively; 95% confidence interval (CI) of difference 12.1 s, 59.1 s; P = 0.008]. There was no significant effect on peak oxygen consumption (placebo, 50.6+/-6.3 ml.min( 1).kg(-1); sumatriptan, 51.7+/-7.6 ml.min(-1).kg(-1)). Sumatriptan administration resulted in decreases in both heart rate (sumatriptan, 188+/-14 beats/min, placebo, 196+/-12 beats/min; 95% CI of difference 12.6, 2.6; P = 0.008) and respiratory exchange ratio (sumatriptan, 1.23+/-0.06; placebo, 1.26+/-0.07; 95% CI of difference 0.05, 0.01; P = 0.01) at peak exercise. There were no significant differences in blood pressure, heart rate or submaximal oxygen consumption between sumatriptan and placebo treatments at any stage of exercise. Thus sumatriptan reduces maximal exercise capacity in normal males. The failure to demonstrate any haemodynamic or cardiorespiratory effect suggests that sumatriptan enhances perception of fatigue by a peripheral mechanism affecting 5 HT modulation. PMID- 10814602 TI - Internal carotid artery haemodynamics in women with polycystic ovaries. AB - Although polycystic ovarian syndrome (PCOS) is associated with risk factors for cerebrovascular disease, the available evidence does not demonstrate increased stroke mortality, and it has been suggested that some protective mechanisms must be operating in these women. Haemodynamic changes have been demonstrated in the pelvic vessels and aorta in this syndrome, but there have been no studies of the cerebral circulation. Since insulin resistance is a central feature of PCOS, it is of interest that cerebral blood flow may be altered in diabetic subjects. The present study was designed to assess internal carotid artery haemodynamics in women with PCOS, asymptomatic women with polycystic ovaries (PCO women) and healthy controls. Mean internal carotid pulsatility index, measured using pulsed Doppler ultrasound under standardized conditions, was significantly lower in 35 PCOS and 15 PCO women than in the 18 controls. This was also shown on multiple regression analyses. Peak systolic velocity did not differ between groups. In view of the difficulty of interpreting pulsatility index in low-impedance vascular beds, the 'back' pressure was calculated and found to be lower in women with polycystic ovaries. This is the first demonstration of lower pulsatility index and back pressure (suggestive of reduced vascular tone) in the cerebral circulation of these women, independent of blood pressure, insulin resistance and other endocrine or metabolic factors. Changes in internal carotid haemodynamics may offset the risk of cerebrovascular disease commonly associated with polycystic ovaries. PMID- 10814601 TI - Effects of partial and total colectomy on mineral and acid-base homoeostasis in the rat: magnesium deficiency, hyperphosphaturia and osteopathy, in the presence of high serum 1,25-dihydroxyvitamin D but normal parathyroid hormone. AB - The effects of colectomy on acid-base status, extra-osseous and bone minerals, calciotropic hormones and bone morphology have not yet been studied. To rectify this, groups of normally fed male rats were subjected to distal (n=11), proximal (n=12) or total (n=12) colectomy. Sham-operated rats (n=12) served as controls. At 112 (+/-2) days after colectomy the following changes were noted: (1) weight gain was delayed; (2) faecal excretion of calcium and phosphorus was normal, whereas that of magnesium was increased; (3) intestinal calcium secretion and absorption of calcium and phosphorus were normal, but magnesium absorption was decreased; (4) urinary excretion of magnesium was also decreased, that of phosphorus was increased, and that of pyridinium and deoxypyridinium tended to be high; (5) the serum levels of ionized magnesium, total calcium, 25-hydroxyvitamin D and parathyroid hormone were normal, while that of 1,25-dihydroxyvitamin D was markedly elevated; and (6) bone magnesium and phosphorus content were decreased, but bone calcium was normal, and thus the bone calcium/phosphorus ratio was high. These abnormalities were associated with moderate metabolic acidosis, as reflected by high urinary ammonium, low citrate and low total CO(2), but normal blood gases. Significant structural abnormalities of bone were not detectable, but trabecular bone tended to show rarefication. Distal colectomy had the least effect, whereas proximal and total colectomies had a distinct effect, on these parameters. It is concluded that colectomy in the rat causes: (1) a syndrome of magnesium deficiency of intestinal origin, compensated metabolic acidosis, urinary phosphorus loss, and high circulating 1,25-dihydroxyvitamin D levels, with the degree depending on the extent of surgical resection; and (2) brittle bones, a feature characteristic of low bone magnesium and more generalized magnesium deficiency. The mechanisms leading to this syndrome are unknown, but altered tissue levels of magnesium and phosphorus may play a key role. PMID- 10814603 TI - Abnormal venous function in patients with homozygous sickle cell (SS) disease and chronic leg ulcers. AB - Chronic leg ulceration is a major cause of morbidity in homozygous sickle cell (SS) disease in Jamaica. These ulcers have features in common with venous ulcers in patients with a normal haemoglobin genotype (AA). Thus we sought to determine whether there is abnormal venous function in the legs of patients with SS disease who have ulcers. Experiments were performed on 15 SS patients with ulcers, and on 15 SS patients and 15 AA subjects with no history of leg ulcers. Changes in venous blood volume of the bottom one-third of the leg induced by venous occlusion and release were studied by air plethysmography, providing indices of segmental venous capacitance (SVC), maximal venous outflow (MVO) and venous emptying time (VET). The changes in volume (ambulatory volume change; AVC) induced by a period of leg exercise were also measured at the ankle (AVCa) and calf (AVCc); venous refilling times at these sites (RTa and RTc respectively) were also measured. Finally, cutaneous red blood cell flux recovery time (FRT) after ankle exercise was assessed by laser Doppler flowmetry. Measurements were also made of haematological variables. SVC, MVO and VET did not differ between the groups, indicating no deep venous obstruction in the SS patients with ulcers. AVCc, AVCa and RTc did not differ among the three subject groups. However, compared with AA subjects, SS patients with ulcers had reduced RTa and FRT. Moreover, RTa and FRT were further shortened in SS patients with ulcers relative to SS patients without ulcers. Since the levels of anaemia were similar in SS patients with and without ulcers, these differences cannot be attributed to differences in arterial flow secondary to anaemia. These results suggest abnormal venous function in SS patients with ulcers, relative to both AA subjects and SS patients without ulcers. We propose that there is incompetence of venous valves draining the ankle region of SS patients with ulcers: the consequent raised venous pressure contributes to the slow healing and, possibly, to the onset of leg ulceration in SS disease. PMID- 10814604 TI - Abnormal thiol group modulation of sodium-lithium countertransport and membrane fluidity is associated with a disturbed relationship between serum triacylglycerols and membrane function in type II diabetes. AB - In essential hypertension and diabetic nephropathy, sodium-lithium countertransport (Na-Li CT) is an inherited marker, subject to metabolic influences, of cardiovascular risk. Studies in Type II diabetes, taking clinical phenotypes as their starting point, are conflicting. We sought to identify Na-Li CT kinetic abnormalities in Type II diabetes, and only subsequently to seek relationships with clinical variables. Na-Li CT kinetics, membrane fluidity and their modulation by thiol proteins were measured in erythrocytes from 38 patients with Type II diabetes and in 16 normal control subjects. In untreated erythrocytes, Na-Li CT kinetics were similar. Thiol protein alkylation with N ethylmaleimide generally caused both V(max) and K(m) to fall, but caused K(m) to rise in erythrocytes from 13 out of 38 diabetic subjects, whose native K(m) was low (P=0. 0013 compared with control). V(max) and serum triacylglycerol levels were related in normal controls (r(s)=0.54, P=0.038) and in diabetic subjects whose K(m) fell after N-ethylmaleimide (n=25, r(s)=0.62, P=0.001). Where the K(m) rose after N-ethylmaleimide, V(max) and triacylglycerol levels were not related (n=13, r(s)=-0.39, P=0.183) and membrane fluidity did not increase after N ethylmaleimide. However, these subgroups were indistinguishable in terms of blood pressure, albuminuria, glycaemia or lipid profiles. Thus abnormalities in the regulation of Na-Li CT and membrane fluidity by key thiol proteins, resembling those seen in essential hypertension and diabetic nephropathy, were apparent in one-third of subjects with Type II diabetes. Membrane abnormalities may indicate a common pathological mechanism. The prognostic significance of Na-Li CT kinetic abnormalities in Type II diabetes must now be confirmed. PMID- 10814605 TI - Perception of bronchoconstriction and bronchial hyper-responsiveness in asthma. AB - The inter-relationship between the perception of bronchoconstriction, bronchial hyper-responsiveness and temporal adaptation in asthma is still a matter of debate. In a total of 52 stable asthmatic patients, 32 without airway obstruction ?forced expiratory volume in 1 s (FEV(1))/vital capacity (VC) 84.1% (S.D. 7.9%), and 20 with airway obstruction [FEV(1)/VC 60% (4%)], we assessed the perception of bronchoconstriction during methacholine inhalation by using: (i) the slope and intercept of the Borg and VAS (Visual Analog Scale) scores against the decrease in FEV(1), expressed as a percentage of the predicted value; and (ii) the Borg and VAS scores at a 20% decrease in FEV(1) from the lowest post-saline level (PB(20)). Bronchial hyper-responsiveness was assessed as the provocative concentration of methacholine causing a 20% fall in FEV(1) (PC(20)FEV(1)). The reduction in FEV(1) was significantly related to the Borg and VAS scores, with values for the group mean slope and intercept of this relationship of 0.13 (S.D. 0.08) and -1.1 (3.02) for Borg, and 1.5 (1.19) and -12.01 (35) for VAS. PB(20) was 3 (1.75) with Borg scores and 34.6 (20.5) with VAS scores. Compared with the subgroup without airway obstruction, the obstructed subgroup exhibited similar slopes, but lower Borg and VAS intercepts. For similar decreases in FEV(1) (5-20% decreases from the lowest post-saline values), the Borg and VAS scores were lower in the non-obstructed than in the obstructed subgroup. PC(20)FEV(1) was significantly related to both Borg PB(20) and VAS PB(20) when considering all patients. When assessing the subgroups, PC(20)FEV(1) was related to Borg PB(20) and VAS PB(20) in the non-obstructed subjects, but not in the obstructed subjects. In neither subgroup was the log of the cumulative dose related to the Borg and VAS scores at the end of the test. We conclude that, unlike in previous studies, the ability to perceive acute bronchoconstriction may be reduced as background airflow obstruction increases in asthma. Bronchial hyper responsiveness did not play a major role in perceived breathlessness in patients without airway obstruction, and even less of a role in patients with obstruction. The cumulative dose of agonist did not appear to influence the perception of bronchoconstriction. PMID- 10814606 TI - Effects of ATP infusion on glucose turnover and gluconeogenesis in patients with advanced non-small-cell lung cancer. AB - Cancer cachexia is associated with elevated lipolysis, proteolysis and gluconeogenesis. ATP infusion has been found to significantly inhibit loss of body weight, fat mass and fat-free mass in patients with advanced lung cancer. The present study was aimed at exploring the effects of ATP on whole-body glucose turnover, alanine turnover and gluconeogenesis from alanine. Twelve patients with advanced non-small-cell lung cancer (NSCLC) were studied 1 week before and during 22-24 h of continuous ATP infusion. After an overnight fast, turnover rates of glucose and alanine, and gluconeogenesis from alanine, were determined using primed constant infusions of ?6, 6-(2)H(2)glucose and ?3-(13)Calanine. Thirteen NSCLC patients and eleven healthy subjects were studied as control groups without ATP infusion. During high-dose ATP infusion (75 microg.min(-1).kg(-1)), glucose turnover was 0.62+/-0.07 mmol.h(-1).kg(-1), compared with 0. 44+/-0.13 mmol.h( 1).kg(-1) at baseline (P=0.04). For gluconeogenesis a similar, but non significant, trend was observed ?baseline, 0.30+/-0.16 mmol.h(-1).kg(-1); during ATP, 0.37+/-0.13 mmol.h(-1).kg(-1) (P=0.08). At lower ATP doses (37-50 microg. min(-1).kg(-1)) these effects were not detected. The relative increase in glucose turnover during ATP infusion compared with baseline showed a significant correlation with the ATP dose (r=0.58, P=0.02). No change in alanine turnover was observed at any ATP dose. The results of this study indicate an increase in glucose turnover during high-dose ATP infusion compared with baseline levels. During high-dose ATP infusion, glucose turnover was similar to that during low dose ATP infusion and to that in control NSCLC patients. Between ATP infusions, however, glucose turnover in patients treated with high-dose ATP was significantly lower than that in the low-dose and control NSCLC patients (P=0.04 and P=0.03 respectively), and similar to that in healthy subjects. This would suggest that repeated high-dose ATP infusions may inhibit glucose turnover between infusion periods. PMID- 10814607 TI - Blood pressure and renal haemodynamic response to salt during the normal menstrual cycle. AB - The purpose of the present study was to evaluate prospectively blood pressure and the renal haemodynamic response to salt during the normal menstrual cycle. A total of 35 healthy normotensive young women not on oral contraceptives were enrolled; 17 were studied in the follicular phase and 18 in the luteal phase of the menstrual cycle. The women in each group were then randomly allocated to receive a low-sodium (40 mmol/day) or a high-sodium (250 mmol/day) diet for a 7 day period in two consecutive menstrual cycles. At the end of each dietary period, 24 h ambulatory blood pressure, urinary sodium excretion, plasma renin activity, plasma catecholamine levels and renal haemodynamics were measured. Our results show that the blood pressure response to salt is comparable during the luteal and the follicular phases of the normal menstrual cycle and is characterized by a salt-resistant pattern. In the kidney, effective renal plasma flow was significantly greater and the filtration fraction lower (P<0.05) after salt loading in women studied in the luteal phase compared with women investigated in the follicular phase. This study thus demonstrates that the female hormone status does not affect the blood pressure response to sodium in young normotensive women. However, in contrast with systemic haemodynamics, the renal response to salt varies during the normal menstrual cycle, suggesting that female sex hormones play a role (direct or indirect) in the regulation of renal haemodynamics. PMID- 10814608 TI - Myocardial infarction with and without reperfusion in sheep: early cardiac and neurohumoral changes. AB - There are few stable and reproducible large-animal models of chronic heart failure produced by ischaemic damage to the myocardium. Here we characterize a novel method of inducing myocardial damage in closed-chest sheep by catheter delivery of thrombogenic coils, and compare this with a newly described open artery model of cardiac injury in sheep. Sham controls were compared with animals subjected to (a) 90 min of coronary artery occlusion/reperfusion by PTCA (percutaneous transluminal coronary angioplasty) balloon, and (b) permanent coronary artery occlusion induced by catheter delivery of thrombogenic coils (seven sheep/group). Both balloon occlusion/reperfusion and permanent coil occlusion resulted in well-defined anteroapical infarcts, as documented by ECG changes, significant rises in creatine kinase (both groups P<0.001) and troponin T (both groups P<0.05), and post-mortem examination. Washout of enzymes was much more rapid in the reperfused group (P<0. 01). Infarction resulted in significant reductions in left ventricular (LV) ejection fraction (both groups P<0.01) and regional wall abnormalities. Ejection fraction 7 days post-coil (21.3+/-4.2%) was significantly lower (P<0.01) than that 7 days post-balloon (38. 8+/-4.5%). Coil induced infarction was associated with acutely reduced arterial pressure (P<0.05), and increases in heart rate (P<0. 05), atrial pressures (P<0.05), plasma brain natriuretic peptide levels (P<0.05) and adrenaline levels (P<0.05). Rises seen in plasma endothelin levels in sham controls were blunted in the coil group (P<0.001). Haemodynamic changes were less marked in the balloon group. In conclusion, restriction of coronary artery occlusion to 90 min results in infarction, but less LV dysfunction with reduced early remodelling, compared with permanent occlusion. Acute changes in biochemical markers, haemodynamics, neurohormones and LV function confirm that these are excellent models of open- and closed-artery myocardial infarction leading to asymptomatic LV dysfunction. PMID- 10814609 TI - The case for an aggressive surgical approach to papillary muscle rupture following myocardial infarction: "From paradise lost to paradise regained". PMID- 10814610 TI - Images in cardiology. Continuous murmur through atrial septal defect. PMID- 10814611 TI - Improving ventricular systolic function: simple messages from complex models. PMID- 10814612 TI - Evaluating aortic dissection: when is coronary angiography indicated? PMID- 10814614 TI - Stamps in cardiology. Transplantation. PMID- 10814613 TI - BNP: soon to become a routine measure in the care of patients with heart failure? PMID- 10814615 TI - Lipid lowering drugs for patients who continue to smoke? PMID- 10814616 TI - Management of cardiogenic shock complicating acute myocardial infarction: towards evidence based medical practice. PMID- 10814618 TI - The left ventricular dysfunction questionnaire (LVD-36): reliability, validity, and responsiveness. AB - OBJECTIVE: To examine the reliability, validity, and responsiveness of a new health status measure (LVD-36) for patients with left ventricular dysfunction which was designed with emphasis on content validity, clarity, brevity, and ease of use. DESIGN: At baseline, patients completed the LVD-36 and a range of measures reflecting general health and disease severity. The LVD-36 was repeated after one week. After six months, it was repeated again, along with a transition question to measure global changes in health. SETTING: Patients were recruited from the cardiology and general medical clinics at a south west London hospital. PATIENTS: 60 patients with chronic left ventricular dysfunction. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Short form 36 questionnaire (SF-36), Minnesota living with heart failure questionnaire (LIhFE), New York Heart Association criteria, and exercise performance and echocardiographic tests. RESULTS: The LVD 36 showed good internal consistency (kappa = 0.95) and repeatability (r(i) = 0.95). Its scores were significantly associated with SF-36 mental and physical component scores (r = -0. 48 and -0.75; p < 0.0001), with exercise capacity (r = 0.52; p < 0. 0001), and with systolic shortening fraction (r = -0.27; p < 0.05). Change in the LVD-36 over six months was associated with change in overall health (F = 5.7; p < 0.001). In tests of validity and responsiveness, the LVD-36 performed similarly to or marginally better than the LIhFE. CONCLUSIONS: The LVD 36 showed a high level of reliability and validity, and appears to measure changes in health. It provides a short, simple, valid, and reliable measure of health status in patients with left ventricular dysfunction. PMID- 10814617 TI - Recreational drug misuse: issues for the cardiologist. PMID- 10814619 TI - Images in cardiology. Cullen's sign after coronary angiography. PMID- 10814620 TI - Measuring the health related quality of life of people with ischaemic heart disease. AB - OBJECTIVES: To inform researchers and clinicians about the most appropriate generic and disease specific measures of health related quality of life for use among people with ischaemic heart disease. METHODS: MEDLINE and BIDS were searched for research papers which contained a report of at least one of the three most common generic instruments or at least one of the five disease specific instruments used with ischaemic heart disease patients. Evidence for the validity, reliability, and sensitivity of these instruments was critically appraised. RESULTS: Of the three generic measures-the Nottingham health profile, sickness impact profile, and short form 36 (SF-36)-the SF-36 appears to offer the most reliable, valid, and sensitive assessment of quality of life. However, a few of the SF-36 subscales lack a sufficient degree of sensitivity to detect change in a patient's clinical condition. According to the best available evidence, the quality of life after myocardial infarction questionnaire should be preferred to the Seattle angina questionnaire, the quality of life index cardiac version, the angina pectoris quality of life questionnaire, and the summary index. Overall, research on disease specific measures is sparse compared to the number of studies which have investigated generic measures. CONCLUSIONS: An assessment of the quality of life of people with ischaemic heart disease should comprise a disease specific measure in addition to a generic measure. The SF-36 and the quality of life after myocardial infarction questionnaire (version 2) are the most appropriate currently available generic and disease specific measures of health related quality of life, respectively. Further research into the measurement of health related quality of life of people with ischaemic heart disease is required in order to address the problems (such as lack of sensitivity to detect change) identified by the review. PMID- 10814621 TI - Health related quality of life and psychological wellbeing in patients with dilated cardiomyopathy. AB - OBJECTIVE: To assess the health related quality of life and psychological wellbeing of patients with dilated cardiomyopathy, and relate these to clinical variables and psychological adjustment. DESIGN: Postal questionnaire survey of 99 adult patients with dilated cardiomyopathy, selected at random from a larger database (60.6% response rate). Assessments included the short form 36 (SF-36) health survey, the hospital anxiety and depression scales, the sleep problems index, and a measure of psychological adjustment to cardiomyopathy. RESULTS: Patients with dilated cardiomyopathy reported significant impairments in physical functioning, role limitations owing to physical and emotional problems, social functioning, mental health, perceptions of general health, sleep, and vitality. Anxiety and depression levels were higher than in population samples. Impairment in several domains of quality of life was associated with low shortening fraction, high left ventricular end diastolic diameter, and the presence of heart failure and mitral regurgitation. Patients with familial cardiomyopathy had less impairment in quality of life than those with non-familial disease. Psychological adjustment scores were associated with several aspects of quality of life and emotional wellbeing. In multivariate analysis, demographic and clinical variables accounted for 0.1-40.7% of the variance in different domains of quality of life, and psychological adjustment scores accounted for an additional 0.5-22.4% of variance. CONCLUSIONS: Patients with dilated cardiomyopathy experience pronounced restrictions in quality of life and psychological wellbeing. These limitations are only partly accounted for by symptoms and the severity of underlying disease. Patients may benefit from efforts to improve psychological adjustment to the condition. PMID- 10814622 TI - Measuring the costs and benefits of heart disease monitoring. AB - OBJECTIVE: To evaluate the costs and benefits of alternative systems of coronary heart disease monitoring in Scotland. DESIGN: An option appraisal was conducted to evaluate the costs and benefits of implementing a coronary heart disease monitoring system. This involved a review of existing Scottish datasets and relevant reports, specification of options, definition and weighting of benefit criteria by key stakeholders, assessment of options by experts, and costing of options. The options were assessed by 33 stakeholders (grouped as cardiologists, patient representatives, general practitioners, public health physicians, and policy makers), plus 13 topic experts. SETTING: Scotland (population 5.1 million). RESULTS: Between group mean benefit weights were: mortality rates and case fatality (10.6), quality of life (9.8), patient function (8.8), hospital activity (7.8), primary care activity (9.25), prescribing (5.72), socioeconomic impact (4.0), risk factors (7.4), prevalence (5.0), incidence (6.0), case registration (6.82), international comparability (4.2), breadth of coverage (8.8), and frequency (5.8). Differences between group weights were significant for prevalence (p = 0.048) and international comparability (p = 0.032). Four monitoring options were identified: a community epidemiology model, based on MONICA (monitoring trends and determinants in cardiovascular disease) study methodology applied to a series of eight representative communities, had the highest benefits, at an average annual discounted cost of approximately pound 360,000; models based on the Australian cardiovascular disease monitoring scheme and on enhanced routine data offered fewer benefits at discounted average annual costs ranging from pound 165,000 to pound 195,000; finally, a coronary heart disease registry modelled on the Scottish Cancer Registry scheme would have had fewer benefits and substantially higher costs than the other options. CONCLUSIONS: The most beneficial coronary heart disease monitoring system is the community epidemiology model, based on MONICA methodology. Option appraisal potentially offers an explicit and transparent methodology for evidence based policy development. PMID- 10814623 TI - Improving the ECG classification of inferior and lateral myocardial infarction by inversion of lead aVR. AB - OBJECTIVE: To assess whether the use of inverted lead aVR (-aVR) would improve the classification of acute inferior or lateral myocardial infarction presenting with ST elevation. DESIGN: Observational study. The presence of >/= 1 mm ST elevation in lead -aVR (derived by manual assessment of ST depression in conventional lead aVR) was determined by a single investigator, blinded to patient outcome. PATIENTS: 173 consecutive patients with chest pain for /= 1 mm in inferior leads (II, III, aVF) or lateral leads (I, aVL, V5, V6), excluding those with anterolateral ST elevation. MAIN OUTCOME MEASURE: Incidence of ST elevation in lead -aVR in patients with inferior or lateral ST elevation, or both. RESULTS: ST elevation in lead -aVR was present in 25 of 136 patients (18%) with inferior but no lateral ST elevation (indicating greater superior involvement) and in three of 11 patients (27%) with lateral but no inferior ST elevation (indicating greater inferior involvement). ST elevation in lead -aVR bridged the gap between inferior and lateral ST elevation in 15 of 25 (60%) patients with inferior and lateral chest lead (V5/V6) ST elevation, and in all patients with inferior and lateral limb lead (I/aVL) ST elevation. The presence of ST elevation in lead -aVR was associated with a larger infarct size as defined by median peak creatine kinase on serial sampling: 1780 v 987 mmol/l; p = 0.021. CONCLUSIONS: Use of lead -aVR improves the ECG classification of acute inferior or lateral acute myocardial infarction and thus may be useful as part of the routine 12 lead ECG assessment of such patients. PMID- 10814625 TI - Images in cardiology. Ventricular pacing and right bundle branch block morphology: diagnosis and management. PMID- 10814624 TI - Arrhythmias in the congenital long QT syndrome: how often is torsade de pointes pause dependent? AB - OBJECTIVE: To determine the frequency and predictors of pause dependent torsade de pointes among patients with the congenital long QT syndrome and spontaneous ventricular tachyarrhythmias. DESIGN: The literature on the "congenital long QT" was reviewed. Articles with illustrations demonstrating the onset of spontaneous polymorphic ventricular arrhythmias in the absence of arrhythmogenic drugs were included. RESULTS: Illustrations of 62 spontaneous episodes of torsade de pointes among patients with congenital long QT syndrome were found in the literature. The majority (74%) of documented arrhythmias were "pause dependent"; 82% of these pauses were longer than the basic cycle length by > 100 ms. Age and sex correlated with the mode of arrhythmia initiation. Arrhythmias in infants (/= 30 kg/m(2)). The index did not predict all cause mortality when examined by quintile. For major ischaemic heart disease (non-fatal or fatal ischaemic heart disease) the relative odds was 1.73 (95% CI 1.21, 2.48) in the top fifth of the distribution (body mass index >/= 24.2 kg/m(2)) compared with the bottom fifth (body mass index <20.1 kg/m(2)). In men with an index >/= 30 kg/m(2) however, the relative odds were 2.03 (95% CI, 1.03, 4.01) for all cause mortality and 2.17 (95% CI, 1.08, 4.34) for major ischaemic heart disease, adjusted for age, smoking habit and social class. When men were recruited to the study, from 1979 to 1983; the mean body mass index had increased to 26.2 (SD 3.6), a mean increase of 3.9 kg/m(2) or 11. 2 kg; 299 men (12.1%) were classified as obese and showed significantly increased relative odds of both all cause mortality (1. 53 (95% CI 1.14, 2.06) and major ischaemic heart disease (1.55 (95% CI 1.13, 2.11)), adjusted for age, smoking habit and social class relative to the non-obese men. The effect of gain in weight from 18 years to recruitment was also examined; all cause mortality showed highest mortality in the fifth of the distribution who experienced weight loss or minimal weight gain. For major ischaemic heart disease an inconsistent, weak trend was shown, the relative odds rising to a maximum of 1.26 (0.89, 1.80) in the top fifth of weight gain compared with the bottom fifth. Weight gain showed strong associations with potential cardiovascular risk factors measured at recruitment; insulin, triglyceride, glucose, diastolic and systolic blood pressure and high density lipoprotein cholesterol. CONCLUSIONS: Body mass at 18 years of age of 30 kg/m(2) or more conferred increased risk for all cause mortality and major ischaemic heart disease during 14 years of follow up of men aged 45 to 59 years. By the baseline examination the prevalence of obesity (body mass index >/=30) had increased from 1.8% to 12.1%; obese men also showed an excess risk of major ischaemic heart disease and overall mortality, but these risks were lower than those predicted from 18 years of age. Weight gain was strongly associated with smoking habit, the greatest weight gain being among ex-smokers and the least among light smokers. Weight gain from 18 years of age to baseline examination showed little relation with subsequent mortality and risk of major ischaemic heart disease when adjusted for age, smoking habit and social class. The lowest mortality rate occurred in the "fifth" of men who gained a mean weight of 16.1 kg. Weight gain is closely associated with some adverse cardiovascular risk factors; in particular with insulin, triglyceride, glucose and diastolic blood pressure. PMID- 10814655 TI - Smoking cessation at the workplace. Results of a randomised controlled intervention study. Worksite physicians from the AIREL group. AB - OBJECTIVES: To compare the effects of a worksite intervention by the occupational physician offering simple advice of smoking cessation with a more active strategy of advice including a "quit date" and extra support. POPULATION: Employees of an electrical and gas company seen at the annual visit by their occupational physicians. CRITERIA END POINTS: Smoking point prevalence defined as the percentage of smokers who were non-smokers at one year. Secondary criteria were the percentage of smokers who stopped smoking for more than six months and the difference in prevalence of smoking in both groups. METHODS: Randomised controlled trial. The unit of randomisation was the work site physician and a random sample of the employees of whom he or she was in charge. The length of the follow up was one year. Each of 30 work site physicians included in the study 100 to 150 employees. RESULTS: Among 504 subjects classified as smokers at baseline receiving simple advice (group A) and 591 the more active programme (group B), 68 (13.5%) in group A and 109 (18. 4%) were non-smokers one year later (p=0.03; p=0.01 taking the occupational physician as the statistical unit and using a non parametric test). Twenty three subjects (4.6%) in group A and 36 (6.1%) in group B (p=0.26) declared abstinence of six months or more. Among non-smokers at baseline, 3.4% in both groups were smokers after one year follow up. The prevalence of smokers did not differ significantly at baseline (32.9% and 32.4%, p=0.75). After the intervention the prevalence of smoking was 30.8% in group A and 28. 7% in group B (p=0.19). An increase of the mean symptoms score for depression in those who quit was observed during this period. CONCLUSIONS: A simple cessation intervention strategy during a mandatory annual examination, targeting a population of smokers independently of their motivation to stop smoking or their health status, showed a 36% relative increase of the proportion of smokers who quit smoking as compared with what can be achieved through simple advice. PMID- 10814656 TI - Plasma vitamin C and food choice in the third Glasgow MONICA population survey. AB - STUDY OBJECTIVE: To determine the contribution of different foods to the estimated intakes of vitamin C among those differing in plasma vitamin C levels, and thereby inform dietary strategies for correcting possible deficiency. DESIGN: Cross sectional random population survey. SETTING: North Glasgow, Scotland, 1992. PARTICIPANTS: 632 men and 635 women, aged 25 to 74 years, not taking vitamin supplements, who participated in the third MONICA study (population survey monitoring trends and determinants of cardiovascular disease). MEASUREMENTS AND MAIN RESULTS: Dietary and sociodemographic information was collected using a food frequency and lifestyle questionnaire. Plasma vitamin C was measured in non fasted venous blood samples and subjects categorised by cut points of 11.4 and 22.7 micromol/l as being of low, marginal or optimal vitamin C status. Food sources of dietary vitamin C were identified for subjects in these categories. Plasma vitamin C concentrations were compared among groups classified according to intake of key foods. More men (26%) than women (14%) were in the low category for vitamin C status; as were a higher percentage of smokers and of those in the older age groups. Intake of vitamin C from potatoes and chips (fried potatoes) was uniform across categories; while the determinants of optimal versus low status were the intakes of citrus fruit, non-citrus fruit and fruit juice. Optimal status was achieved by a combined frequency of fruit, vegetables and/or fruit juice of three times a day or more except in older male smokers where a frequency greater than this was required even to reach a marginal plasma vitamin C level. CONCLUSION: Fruit, vegetables and/or fruit juice three or more times a day increases plasma vitamin C concentrations above the threshold for risk of deficiency. PMID- 10814657 TI - Alcohol consumption in the Baltic Republics. AB - STUDY OBJECTIVES: Premature mortality associated with alcohol intake is of particular concern in several countries of the former Soviet Union. This study explored self reported alcohol consumption (beer, wine, spirits) and its determinants in the Baltic Republics. DESIGN: Cross sectional surveys conducted in 1997. SETTINGS: Estonia, Latvia and Lithuania. PARTICIPANTS: Representative samples of adults age 19-64 (Estonia n=2010; Latvia n=2258; Lithuania n=2139). METHODS: Between country differences in the frequency of alcohol intake were estimated. The odds of consuming alcohol weekly according to socioeconomic characteristics (age, ethnicity, rural/urban area, education, income) were calculated using multiple logistic regression analyses, adjusting for all variables simultaneously. MAIN RESULTS: The proportion of respondents consuming alcohol weekly varied by country (p<0.001) (men: Estonia=61% Latvia=41% Lithuania=55%; women: Estonia=26% Latvia=8% Lithuania=14%). Within each country, this proportion decreased with age in both sexes (p<0. 001), and increased with income in women (p<0.01). In Estonia, the odds of drinking alcohol weekly was significantly lower in respondents of Russian than of Estonian ethnicity (odds ratios (OR) and 95% confidence intervals (95%CI): men=0.51 (0.36, 0.71); women=0. 57 (0.39, 0.81)). In Lithuania, the odds was higher in highly educated men than in those with a low education level (OR=1.48 (1.01, 2.17)). Daily alcohol intake was higher in Estonia than in the other countries, as was the percentage of respondents drinking heavily (equivalent to 80 g/day). CONCLUSIONS: Approximately half the men and one in six women in the Baltic States reported consuming alcohol at least weekly. Age and income were the strongest and most consistent correlates of the likelihood of consuming alcohol weekly. Ethnic differences were observed only in Estonia. PMID- 10814658 TI - Ecological effects in multi-level studies. AB - Multi-level research that attempts to describe ecological effects in themselves (for example, the effect on individual health from living in deprived communities), while also including individual level effects (for example, the effect of personal socioeconomic disadvantage), is now prominent in research on the socioeconomic determinants of health and disease. Such research often involves the application of advanced statistical multi-level methods. It is hypothesised that such research is at risk of reaching beyond an epidemiological understanding of what constitutes an ecological effect, and what sources of error may be influencing any observed ecological effect. This paper aims to present such an epidemiological understanding. Three basic types of ecological effect are described: a direct cross level effect (for example, living in a deprived community directly affects individual personal health), cross level effect modification (for example, living in a deprived community modifies the effect of individual socioeconomic status on individual health), and an indirect cross level effect (for example, living in a deprived community increases the risk of smoking, which in turn affects individual health). Sources of error and weaknesses in study design that may affect estimates of ecological effects include: a lack of variation in the ecological exposure (and health outcome) in the available data; not allowing for intraclass correlation; selection bias; confounding at both the ecological and individual level; misclassification of variables; misclassification of units of analysis and assignment of individuals to those units; model mis-specification; and multicollinearity. Identification of ecological effects requires the minimisation of these sources of error, and a study design that captures sufficient variation in the ecological exposure of interest. PMID- 10814659 TI - Inequalities in health. Analytic approaches based on life expectancy and suitable for small area comparisons. AB - STUDY OBJECTIVE: Simple measures of inequalities in health are proposed to facilitate the work of health policy makers and to build on the understanding of health differences between populations. In addition, it is aimed to make these measures applicable for comparisons of small populations and subgroups. METHODS: Inequalities in health or health deficiencies were quantified as the difference between the life expectancy of the subgroup of interest and that of the national population. Health deficiencies were divided into disease specific components by partial application of cause eliminated life table methods. To manage small numbers and to depict time trends, locally weighted regression smoothing was applied. Confidence intervals were constructed through Monte Carlo simulations. APPLICATIONS AND COMPARISONS: The proposed approaches were applied to the health situation in Cape Breton County, Nova Scotia, Canada, and disclosed the significance of different diseases and distinct patterns between communities. The proposed measures were also compared with the traditionally used standardised mortality rates and ratios. Here, the proposed measures appeared beneficial in that they are easier to comprehend and that they provide time trends and more robust estimates. CONCLUSIONS: The above advantages make the proposed approaches beneficial to health policy makers and epidemiologists. The approaches may also be incorporated in economic evaluations as well as in more sophisticated public health models. PMID- 10814660 TI - Validity of rapid estimates of household wealth and income for health surveys in rural Africa. AB - STUDY OBJECTIVE: To test the validity of proxy measures of household wealth and income that can be readily implemented in health surveys in rural Africa. DESIGN: Data are drawn from four different integrated household surveys. The assumptions underlying the choice of wealth proxy are described, and correlations with the true value are assessed in two different settings. The expenditure proxy is developed and then tested for replicability in two independent datasets representing the same population. SETTING: Rural areas of Mali, Malawi, and Cote d'Ivoire (two national surveys). PARTICIPANTS: Random sample of rural households in each setting (n=275, 707, 910, and 856, respectively). MAIN RESULTS: In both Mali and Malawi, the wealth proxy correlated highly (r>/=0.74) with the more complex monetary value method. For rural areas of Cote d'Ivoire, it was possible to generate a list of just 10 expenditure items, the values of which when summed correlated highly with expenditures on all items combined (r=0.74, development dataset, r=0. 72, validation dataset). Total household expenditure is an accepted alternative to household income in developing country settings. CONCLUSIONS: It is feasible to approximate both household wealth and expenditures in rural African settings without dramatically lengthening questionnaires that have a primary focus on health outcomes. PMID- 10814661 TI - Setting health priorities in a Swiss canton: what do different methods tell us? AB - STUDY OBJECTIVE: Despite excellent mortality indicators, there is clear evidence that the health status of the population of Geneva could be improved if more attention and resources were devoted to prevention strategies. To identify a set of robust health priorities an original approach was used triangulating results between three methods. METHODS: The study calculated potential years of life lost, disability adjusted years of life lost, and conducted a Delphi survey to gather the opinion of health professionals and the general public. MAIN RESULTS: Several health conditions were unanimously selected by all three methods as top priorities: cardiovascular diseases, AIDS, respiratory cancer, breast cancer for women, suicide and traffic accidents. In addition, two determinants-alcohol abuse and tobacco abuse-for which a clear conceptual link could be established between all methods were chosen. Connections between priorities identified through the DALY and the Delphi method lead to further inclusion of chronic back pain and depression. Some issues solely identified through the Delphi survey were included as they were consistently considered important by professionals and the lay public alike-violence in the family, unemployment, social exclusion. CONCLUSIONS: These results indicate that health priorities, and by extension health care priorities, would benefit from using a mix of quantitative and qualitative research methods. The triangulation of results allows for a broader perspective and makes results more acceptable. PMID- 10814662 TI - Low immunisation uptake: is the process the problem? AB - OBJECTIVE: To examine mothers' satisfaction with the process of immunisation and its possible contribution to suboptimal immunisation uptake. DESIGN: In depth interviews with mothers. SETTING: Two Community Care Areas, Dublin city, Ireland. PARTICIPANTS: In depth interviews of 23 mothers of children 1-2 years old, recruited purposively from a birth cohort born in 1994. MAIN RESULTS: Mothers preferred general practice to Health Centre immunisation (11:5) for predominantly emotional compared with practical reasons (4:1). Health Centre immunisation was seen, at times, as unacceptably rough and inhuman. Many mothers experienced severe emotional distress at the prospect of inflicting the pain of immunisation on their babies. The non-empathic stance of some immunising doctors was unacceptable to mothers. They valued attempts by health professionals to acknowledge the pain of immunisation and to engage with their baby. Adverse experiences contributed to deferral of future visits and to defaulting behaviour. CONCLUSIONS: Low empathy mass immunisation in clinic type settings may be unacceptable to mothers in the 1990s, and may in part explain suboptimal uptake in health care systems that use such clinics. PMID- 10814663 TI - Retinoids and demethylating agents- looking for partners. PMID- 10814664 TI - Metabolically activated carcinogens and mutations in the p53 tumor suppressor gene in lung cancer. PMID- 10814665 TI - Why were NIH adverse event reporting guidelines misunderstood? PMID- 10814667 TI - Detection network gives early cancer tests a push. PMID- 10814668 TI - Some promising biomarkers for cancer. PMID- 10814669 TI - DNA methylation: what is its role in carcinogenesis? PMID- 10814672 TI - Telling cells to die: apoptosis research takes off. PMID- 10814674 TI - Cancer risk and the ATM gene: a continuing debate. AB - Deficiencies in the ability of cells to sense and repair damage in individuals with rare genetic instability syndromes increase the risk of developing cancer. Ataxia-telangiectasia (A-T), such a condition, is associated with a high incidence of leukemia and lymphoma that develop in childhood. Although A-T is an autosomal recessive disorder, some penetrance appears in individuals with one mutated ATM gene (A-T carriers), namely, an increased risk of developing breast cancer. The gene mutated in A-T, designated ATM, is homologous to several DNA damage recognition and cell cycle checkpoint control genes from other organisms. Recent studies suggest that ATM is activated primarily in response to double strand breaks, the major cytotoxic lesion caused by ionizing radiation, and can directly bind to and phosphorylate c-Abl, p53, and replication protein A (RPA). Analysis of ATM mutations in patients with A-T or with sporadic non-A-T cancers has suggested the existence of two classes of ATM mutation: null mutations leading to A-T and dominant negative missense mutations predisposing to cancer in the heterozygous state. Studies with A-T mouse models have helped determine the basis of lymphoid tumorigenesis in A-T and have shown that ATM plays a critical role in maintaining genetic stability by ensuring high-fidelity execution of chromosomal events. Thus, ATM appears to act as a caretaker of the genome. PMID- 10814675 TI - Targeting of lung cancer mutational hotspots by polycyclic aromatic hydrocarbons. AB - BACKGROUND: Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in combustion products of organic matter, including cigarette smoke. Metabolically activated diol epoxides of these compounds, including benzo[a]pyrene diol epoxide (B[a]PDE), have been suggested as causative agents in the development of lung cancer. We previously mapped the distribution of B[a]PDE adducts within the p53 tumor suppressor gene (also known as TP53), which is mutated in 60% of human lung cancers, and found that B[a]PDE adducts preferentially form at lung cancer mutational hotspots (codons 154, 157, 158, 245, 248, and 273). Other PAHs may be important in lung cancer as well. METHODS: Here we have mapped the distribution of adducts induced by diol epoxides of additional PAHs: chrysene (CDE), 5 methylchrysene (5-MCDE), 6-methylchrysene (6-MCDE), benzo[c]phenanthrene (B[c]PDE), and benzo[g]chrysene (B[g]CDE) within exons 5, 7, and 8 of the p53 gene in human bronchial epithelial cells. RESULTS: CDE exposure produced only low levels of adducts. Exposure of cells to the other activated PAHs resulted in DNA damage patterns similar to those previously observed with B[a]PDE but with some distinct differences. 5-MCDE, 6-MCDE, B[g]CDE, and B[c]PDE efficiently induced adducts at guanines within codons 154, 156, 157, 158, and 159 of exon 5, codons 237, 245 and 248 of exon 7, and codon 273 of exon 8, but the relative levels of adducts at each site varied for each compound. B[g]CDE, B[c]PDE, and 5-MCDE induced damage at codon 158 more selectively than 6-MCDE or B[a]PDE. The sites most strongly involved in PAH adduct formation were also the sites of highest mutation frequency (codons 157, 158, 245, 248, and 273). CONCLUSION: The data suggest that PAHs contribute to the mutational spectrum in human lung cancer. PMID- 10814676 TI - Cancer surveillance series: changing patterns of cutaneous malignant melanoma mortality rates among whites in the United States. AB - BACKGROUND: Mortality from melanoma among whites is still increasing in the United States. In this study, we describe the changing patterns of melanoma mortality rates among whites by demographic factors and geography and further assess the relationship between the geographic patterns and the UV radiation (UV B) level. METHODS: Age-adjusted incidence and mortality rates were computed by use of the 1970 U.S. population standard. Annual percent changes of mortality were estimated by fitting regression lines to the logarithm of rates. The relationships between melanoma mortality rates and UV-B level over time were assessed by weighted regressions. All statistical tests were two-sided. RESULTS: From 1950-1954 through 1990-1994, melanoma mortality rates increased by 191% and 84% among males and females, respectively. Mortality rates peaked in the 1930 through 1950 birth cohorts for females and in the 1935 through 1950 birth cohorts for males. In the 1950 through 1969 study period, melanoma mortality rates showed a strong North-South gradient, but the gradient weakened in recent periods. The absolute change in mortality for a 10% increase in UV-B among females decreased from 0.08 additional deaths per 100 000 person-years in 1950-1959 to 0.01 additional deaths in 1990-1995. In contrast, the absolute change in mortality among males showed little change over time; additional deaths increased from 0.11 to 0.12 per 100 000 person-years. CONCLUSIONS: Melanoma mortality in the United States reflects the complex interplay of UV radiation levels in each geographic region, the sun-protection behaviors of each generation of males and females in childhood and adulthood, the geographic mobility of the population, and the risk awareness and early detection. PMID- 10814677 TI - Human papillomavirus DNA testing for cervical cancer screening in low-resource settings. AB - BACKGROUND: In many low-resource settings, there are barriers to cytologic screening for cervical cancer. This study evaluates human papillomavirus (HPV) DNA testing as an alternative screening method. METHODS: Cervical samples from 2944 previously unscreened South African women aged 35-65 years were tested for high-risk types of HPV with the use of the Hybrid Capture I (HCI) assay. Women also had a Pap smear, direct visual inspection of the cervix, and Cervicography(TM). Women positive on any screening test were referred for colposcopy. Samples from women with biopsy-confirmed, low-grade squamous intraepithelial lesions (SILs) (n = 95), high-grade SILs (n = 74), or invasive cervical cancer (n = 12) and a random sample of women with no cervical disease (n = 243) were retested for HPV DNA with the use of the more sensitive Hybrid Capture II (HCII) assay. All P values are two-sided. RESULTS: High-risk HPV DNA was detected in 73.3% and 88.4% of 86 women with high-grade SIL or invasive cancer and in 12.2% of 2680 and 18.1% of 243 women without evidence of cervical disease, with the use of the HCI and HCII assays, respectively. HPV DNA testing with the HCII assay was more sensitive than cytology for detecting high-grade SIL and invasive cancer (McNemar's test, P =.04), and testing with the HCI assay was of equivalent sensitivity (P =.61). Cytology had a statistically significantly better specificity (96.8%) than either the HCI assay (87.8%) or the HCII assay (81.9%) (P<.01). Receiver operating characteristic curves identified test cutoff values that allow HPV DNA testing to identify 57% of women with high-grade SIL or cancer, while classifying less than 5% of women with no cervical disease as HPV DNA positive. CONCLUSIONS: HPV DNA testing has a sensitivity equivalent to, or better than, that of cytology. Since HPV DNA testing programs may be easier to implement than cytologic screening, HPV testing should be considered for primary cervical cancer screening in low-resource settings. PMID- 10814678 TI - Methylation and silencing of the retinoic acid receptor-beta2 gene in breast cancer. AB - BACKGROUND: A growing body of evidence supports the hypotheses that the retinoic acid receptor beta2 (RAR-beta2) gene is a tumor suppressor gene and that the chemopreventive effects of retinoids are due to induction of RAR-beta2. RAR-beta2 expression is reduced in many malignant tumors, and we examined whether methylation of RAR-beta2 could be responsible for this silencing. METHODS: RAR beta2 expression was studied by reverse transcription-polymerase chain reaction (RT-PCR) analysis in eight breast cancer cell lines that were either treated with the demethylating agent 5-aza-2'-deoxycytidine and subsequently with all-trans retinoic acid (ATRA) or left untreated. Sodium bisulfite genomic sequencing was used to determine the locations of 5-methylcytosines in the RAR-beta2 genes of three of these cell lines. In 16 breast cancer biopsy specimens and non neoplastic breast tissue, methylation-specific PCR was used to determine the methylation status of RAR-beta2, and, in 13 of the specimens, RT-PCR analysis was used to detect RAR-beta2 expression. RESULTS: Cell lines SK-BR-3, T-47D, ZR-75-1, and MCF7 exhibited expression of RAR-beta2 only after demethylation and treatment with ATRA. The first exon expressed in the RAR-beta2 transcript was methylated in cell lines ZR-75-1 and SK-BR-3. Six breast cancer specimens showed methylation in the same region of the gene. No expression of RAR-beta2 was found in any grade III lesion. An inverse association between methylation and gene expression was found in all grade II lesions. The RAR-beta2 gene from non-neoplastic breast tissue was unmethylated and expressed. CONCLUSIONS: Methylation of the RAR-beta2 gene may be an initial step in breast carcinogenesis; treatment of cancer patients with demethylating agents followed by retinoic acid may offer a new therapeutic modality. PMID- 10814679 TI - Dietary fat, fat subtypes, and breast cancer in postmenopausal women: a prospective cohort study. AB - BACKGROUND: The intake of total dietary fat and of certain fat subtypes has been shown to be strongly associated with breast cancer in international comparisons and in animal experiments. However, observational epidemiologic studies have generally reported either weak positive or no associations. To extend the prospective epidemiologic evidence on this question, we examined the association between adult dietary intake of fat, fat subtypes, and breast cancer in a large, prospective cohort of postmenopausal women. METHODS: Participants were selected from a national breast cancer mammography screening program conducted from 1973 through 1981 at 29 centers throughout the United States. From 1987 through 1989, 40022 postmenopausal women satisfactorily completed a mailed, self-administered questionnaire that included a 60-item National Cancer Institute/Block food frequency questionnaire. Women were then followed for an average of 5.3 years; 996 women developed breast cancer. Risk was assessed by use of Cox proportional hazard regression, with age as the underlying time metric. All statistical tests were two-sided. RESULTS: Compared with women in the lowest quintile (Q1) of percentage of energy from total fat, the adjusted risk ratio (RR) and 95% confidence interval (CI) for women in the highest quintile (Q5) was 1.07 (95% CI = 0.86-1.32). In analyses stratified by history of benign breast disease (BBD), a positive association was observed among only women with no history of BBD (RR (Q5 versus Q1) = 2.20; 95% CI = 1.41-3.42; test for trend, P =.0003). The increased risk in these women appeared to be attributable to unsaturated fat intake and oleic acid in particular. CONCLUSIONS: In this study, there was no overall association between fat intake during adulthood and breast cancer risk; however, among women with no history of BBD, there appeared to be a positive association between total and unsaturated fat intake and breast cancer risk. PMID- 10814680 TI - Risk of breast cancer in prematurely born women. PMID- 10814682 TI - Re: It's time for a change: cigarette smokers deserve meaningful information about their cigarettes. PMID- 10814684 TI - Re: population attributable risk for breast cancer: diet, nutrition, and physical exercise. PMID- 10814687 TI - Empirically supported treatments in pediatric psychology: nocturnal enuresis. AB - OBJECTIVE: To review the medical and psychological literature concerning enuresis treatments in light of the Chambless criteria for empirically supported treatment. METHOD: A systematic search of the medical and psychological literature was performed using Medline and Psychlit. RESULTS: Several review studies and numerous well-controlled experiments have clearly documented the importance of the basic urine alarm alone as a necessary component in the treatment of enuresis or combined with the "Dry-Bed Training" intervention, establishing them as "effective treatments." Other multicomponent behavioral interventions that also include the urine alarm such as "Full Spectrum Home Training" have further improved the outcome for bed-wetters, but are classified as "probably efficacious" at this time because independent researchers have not replicated them. Less rigorously examined approaches that focus on improving compliance with treatment or include a "cognitive" focus (i.e., hypnosis) warrant further study. CONCLUSIONS: We recommend a "biobehavioral" perspective in the assessment and treatment of bed-wetting and suggest that combining the urine alarm with desmopressin offers the most promise and could well push the already high success rates of conditioning approaches closer to 100%. Much important work is yet to be completed that elucidates the mechanism of action for the success of the urine alarm and in educating society about its effectiveness so that its availability is improved. PMID- 10814690 TI - Empirically supported treatments in pediatric psychology: constipation and encopresis. AB - OBJECTIVE: To review the empirical research examining behavioral and medical treatments for constipation and fecal incontinence. METHOD: Sixty-five articles investigating intervention efficacy were identified and reviewed. Twenty-three of the studies were excluded because they were case studies or were less well controlled single-case designs. The intervention protocol for each study was identified and coded, with studies employing the same interventions matched and evaluated according to the Chambless criteria. RESULTS: From the literature base to date, no well-established interventions have emerged. However, four probably efficacious treatments and three promising interventions were identified. Two different medical interventions plus positive reinforcement fit the criteria for the probably efficacious category (one with fiber recommendation and one without). Three biofeedback plus medical interventions fit efficacy category criteria: one probably efficacious for constipation with abnormal defecation dynamics (full medical intervention plus biofeedback for paradoxical contraction), and two fit the promising intervention criteria for constipation and abnormal defecation dynamics (full medical intervention plus biofeedback for EAS strengthening, correction of paradoxical contraction and home practice; and biofeedback focused on correction of paradoxical contraction, medical intervention without fiber recommendation, and positive reinforcement). Two extensive behavioral interventions plus medical intervention also met efficacy criteria for constipation plus incontinence (medical intervention without laxative maintenance plus positive reinforcement, dietary education, goal setting, and skills building presented in a small-group format fits criteria for a promising intervention; and positive reinforcement and skills building focused on relaxation of the EAS during defecation, but without biofeedback, plus medical intervention meets the probably efficacious criteria). CONCLUSIONS: A discussion of the current weaknesses in this research area follows. Specific recommendations for future research are made including greater clarity in treatment protocol and sample descriptions, reporting cure rates rather than success rates, utilization of adherence checks, and investigation of potential differential outcomes for subgroups of children with constipation and incontinence. PMID- 10814692 TI - Social support and personal models of diabetes as predictors of self-care and well-being: a longitudinal study of adolescents with diabetes. AB - OBJECTIVES: To examine whether peer support and illness representation mediate the link between family support, self-management and well-being. METHOD: Fifty two adolescents (12-18 years old) with Type I diabetes were recruited and followed over 6 months, completing assessments of self-management, well-being, and social support. RESULTS: Perceived impact of diabetes and supportive family and friends were prospectively predictive of participants' well-being measures. Although support from family and friends was predictive of better dietary self care, this relationship was mediated by personal model beliefs. In particular, beliefs about the effectiveness of the diabetes treatment regimen to control diabetes was predictive of better dietary self-care. CONCLUSIONS: Both friends and family are important to support adolescents as they live with and manage their diabetes. Personal models of diabetes are important determinants of both dietary self-care and well-being. In addition, personal models may serve to mediate the relationship between social support and dietary behavior. PMID- 10814693 TI - Pain-sensitive temperament: does it predict procedural distress and response to psychological treatment among children with cancer? AB - OBJECTIVE: To evaluate the relationship between pain sensitivity and children's distress during lumbar punctures (LPs), and whether pain sensitivity functions as a moderator of children's responses to a psychological intervention aimed at reducing LP distress. METHOD: Fifty-five children with acute lymphoblastic leukemia (ages 3 to 18) and their parents completed a questionnaire measure of pain sensitivity. Self-report, physiological, and observed measures of distress were collected during the study baseline LP. Children were then randomized into a psychological intervention or an attention control group. Postintervention and follow-up LPs were observed. RESULTS: Higher levels of pain sensitivity were associated with greater anxiety and pain, both prior to and during the LP. Preliminary analyses indicated that pain sensitivity moderated the effects of intervention on distress. Children who were more pain-sensitive and who received no intervention showed greater increases in LP distress over time. In contrast, children who were more pain-sensitive and who received intervention showed greater decreases in LP distress over time. CONCLUSIONS: A measurement of pain sensitivity may be useful in pediatric oncology settings for effectively targeting pain-vulnerable children for psychological intervention. Preliminary analyses indicate that an empirically-supported intervention for procedural distress is efficacious for those children who are most pain-sensitive. PMID- 10814694 TI - Brief report: psychological symptoms in healthy female siblings of adolescents with and without chronic conditions. AB - OBJECTIVE: To examine the psychological impact of having a sibling with a chronic condition on healthy adolescent females and to explore the potential moderating role of birth order on this relationship. METHOD: We compared selected Brief Symptom Index subscales (anxiety, depression, interpersonal sensitivity, hostility) and global severity scores (GSI) in two groups of healthy, inner-city female adolescents matched for sibling age, gender, birth order, and age spacing: 34 sisters of males and females ages 13-19 years with chronic health conditions (ILLSIBS) and 34 sisters of males and females in the same age range without conditions (WELLSIBS). RESULTS: ILLSIBS generally had more symptoms than WELLSIBS. MANOVA yielded significant three-way interactions of sibling illness status, birth order, and gender for the anxiety, hostility, and GSI. A similar pattern was nonsignificant for the two other subscales. Among younger sisters in general and among older sisters of males only, ILLSIBS had higher scores; however, ILLSIBS who were older sisters of females did not differ significantly in symptom levels from the comparable group of WELLSIBS. CONCLUSIONS: Psychological symptoms in sisters of inner-city, male and female adolescents are related to sibling health status. However, the combination of sibling gender and birth order may modify this relationship and should be considered when evaluating psychological risk or designing interventions. PMID- 10814695 TI - Identification and purification of cytolytic antibodies directed against O acetylated sialic acid in childhood acute lymphoblastic leukemia. AB - Sialic acids typically present as terminal sugars of oligo-saccharides are reported to be modified by O-acetylation at the C-9 position on lymphoblasts of childhood acute lymphoblastic leukemia (ALL) patients (Sinha et al., 1999a, Leukaemia, 13, 119-125). We now report high titers of IgG antibodies directed against O-acetylated derivatives of sialic acids (O-AcSA) in serum of ALL patients. These antibodies were purified using bovine submaxillary mucin (BSM) and the IgG distribution was confined to IgG(1)and IgG(2)subclasses; their binding was totally abolished with de-O-acetylation confirming their specificity towards O-AcSA determinants. Flow cytometry demonstrated binding of these antibody fractions to peripheral blood mononuclear cells (PBMC) of both T- and B ALL patients having increased cell surface 9-O-AcSA determinants. Western blotting of membranes derived from PBMC of ALL patients confirmed binding of the antibody to O-acetylated sialoglycoconjugates corresponding to 144, 135, 120, 90, and 36 kDa whereas binding to PBMC from normal individuals corresponded to 144 and 36 kDa. Specificity of the antibody fraction towards 9-O-AcSA was substantiated by hemagglutination and hemagglutination-inhibition assays. The antibody purified from ALL serum selectively mediates complement dependent cytolysis of lymphoblasts expressing O-AcSAs and thereby possibly confers passive protection. The enhanced anti O-AcSA antibody levels allowed for development of a serodiagnostic assay (BSM-ELISA) specific for ALL. Minimal crossreactivity was observed with other hematological disorders like acute myeloid leukemia (n = 16), chronic myeloid leukemia (n = 6), chronic lymphocytic leukemia (n = 7) and non Hodgkin's lymphoma (n = 3) as well as normal healthy individuals (n = 28). The BSM-ELISA therefore provides a simple, noninvasive alternative diagnostic approach for ALL and merits clinical consideration. PMID- 10814696 TI - High-mannose-type oligosaccharides from human placental arylsulfatase A are core fucosylated as confirmed by MALDI MS. AB - Despite numerous studies on arylsulfatase A, the structure of its glycans is not well understood. It has been shown that the concentration of arylsulfatase A increases in the body fluids of patients with some forms of cancer, and the carbohydrate component of arylsulfatase A synthesized in tumor tissues and transformed cells undergoes increased sialylation, phosphorylation and sulfation. To understand the significance of any changes in the glycosylation of arylsulfatase A in cancer, it is important to know the structure of its carbohydrate component in normal tissue. In the present study we have analyzed carbohydrate moieties of human placental arylsylfatase A using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting on Immobilon P and on-blot deglycosylation using PNGase F for glycan release. Profiles of N-glycans were obtained by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Oligosaccharides were sequenced using specific exoglycosidases, and digestion products were analyzed by MALDI MS and the computer matching of the resulting masses with those derived from a sequence database. Fifty picomoles (6 microg) of arylsulfatase A applied to the gel were sufficient to characterize its oligosaccharide content. The results indicated that human placental arylsulfatase A possesses only high mannose-type oligosaccharides, of which almost half are core fucosylated. In addition, there was a minor species of high-mannose-type glycan bearing six mannose residues with a core fucose. This structure was not expected since high mannose-type oligosaccharides basically have not been recognized as a substrate for the alpha1,6-fucosyltransferase. PMID- 10814697 TI - Characterization of the interaction between galectin-1 and lymphocyte glycoproteins CD45 and Thy-1. AB - Galectin-1 is a sugar binding protein specific for beta-galactosides and not requiring metal ions for binding activity. It exists as a soluble protein which forms a noncovalent homodimer and is expressed with a broad tissue distribution. Recently, galectin-1 has been shown to play a possible role in the immune system mediating apoptosis of activated T cells with indirect evidence suggesting that galectin-1 interacts with the heavily glycosylated, transmembrane, protein phosphotyrosine phosphatase CD45. The interaction of galectin-1 with purified lymphocyte cell surface proteins was studied using surface plasmon resonance in a BIAcoretrade mark. Galectin-1 was shown to bind CD45 and Thy-1 in a carbohydrate dependent manner. Several galectin-1 molecules could bind each CD45 molecule. The dissociation constant of dimeric galectin-1 binding to CD45 was measured at approximately 5 microM, indicating the concentration at which cross-linking of cell surface glycoproteins by galectin-1 would occur. A possible role for galectin-1 in the organization of cell surface glycoproteins is discussed. PMID- 10814698 TI - Massive in vitro synthesis of tagged oligosaccharides in 1-benzyl-2-acetamido-2 deoxy-alpha-D-galactopyranoside treated HT-29 cells. AB - Permanent exposure of differentiated HT-29 cells to the sugar analogue, 1-benzyl 2-acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAcalpha- O -bn) leads to marked effects upon the phenotypic properties of mucin-secreting or enterocyte like HT-29 cells: an inhibition in the secretion of mucins, a blockade in the apical targeting of membrane brush border glycoproteins and a swelling of cells with intracellular accumulation of numerous vesicles. Folch extraction and partition of treated enterocyte-like HT-29 cells revealed a very important accumulation of orcinol and/or resorcinol reactive material in the upper phase (usually containing gangliosides), as compared with untreated HT-29 cells and with treated and untreated Caco-2 cells. Structural analysis indicated the accumulation of a series of GalNAcalpha- O -bn derived oligosaccharides, most of them with the common core Galbeta1-3GalNAcalpha- O -bn. These oligosaccharides contained residues of GlcNAc, Gal, Neu5Ac, or Fuc. In particular, the tagged sialyl-Lewis(x)was identified, as well as more complex sialylated derivatives, including the sialyl-Lewis(x)substituted by an additional Neu5Ac residue. The benzylated oligosaccharides were not significantly detected in the culture medium except for Galbeta1-3GalNAcalpha- O -bn. Upon reversion of the treatment, these derivatives dis-appeared from the cells within few days, however were not recovered as such in the culture medium. Intracellular degradation occurred with desialylation and defucosylation as the first steps. The spectacular accumulation of benzylated oligosaccharides in HT-29 cell, permanently exposed to GalNAcalpha- O -bn very likely plays an important role in the alterations of cellular processes previously described in this cell line. The HT-29 cell culture system also appears to be an efficient source of several tagged oligosaccharides. PMID- 10814699 TI - A novel role for nitric oxide in the endogenous degradation of heparan sulfate during recycling of glypican-1 in vascular endothelial cells. AB - We show here that the endothelial cell-line ECV 304 expresses the heparan sulfate proteoglycan glypican-1. The predominant cellular glycoform carries truncated side-chains and is accompanied by heparan sulfate oligosaccharides. Treatment with brefeldin A results in accumulation of a glypican proteoglycan with full size side-chains while the oligosaccharides disappear. During chase the glypican proteoglycan is converted to partially degraded heparan sulfate chains and chain truncated proteoglycan, both of which can be captured by treatment with suramin. The heparan sulfate chains in the intact proteoglycan can be depolymerized by nitrite-dependent cleavage at internally located N-unsubstituted glucosamine moieties. Inhibition of NO-synthase or nitrite-deprivation prevents regeneration of intact proteoglycan from truncated precursors as well as formation of oligosaccharides. In nitrite-deprived cells, formation of glypican proteoglycan is restored when NO-donor is supplied. We propose that, in recycling glypican-1, heparan sulfate chains are cleaved at or near glucosamines with unsubstituted amino groups. NO-derived nitrite is then required for the removal of short, nonreducing terminal saccharides containing these N-unsubstituted glucosamine residues from the core protein stubs, facilitating re-synthesis of heparan sulfate chains. PMID- 10814700 TI - Conformational behavior of hyaluronan in relation to its physical properties as probed by molecular modeling. AB - Hyaluronan (HA) is a linear charged polysaccharide whose structure is made up of repeating disaccharide units. Apparently conflicting reports have been published about the nature of the helical structure of HA in the solid state. Recent developments in the field of molecular modeling of polysaccharides offer new opportunities to reexamine the structural basis underlying the formation and stabilization of ordered structures and their interactions with counterions. The conformational spaces available and the low energy conformations for the disaccharide, trisaccharide, and tetrasaccharide segments of HA were investigated via molecular mechanics calculations using the MM3 force field. First, the results were used to access the configurational statistics of the corresponding polysaccharide. A disordered chain having a persistence length of 75 A at 25 degrees C is predicted. Then, the exploration of the stable ordered forms of HA led to numerous helical conformations, both left- and right-handed, having comparable energies. Several of these conformations correspond to the experimentally observed ones and illustrate the versatility of the polysaccharide. The double stranded helical forms have also been explored and theoretical structures have been compared to experimentally derived ones. PMID- 10814701 TI - Minimal catalytic domain of N-acetylglucosaminyltransferase V. AB - UDP-GlcNAc: Manalpha1-6Manbeta-R beta1-6 N-acetylglucosaminyltransferase V (EC 2.4.1.155, GlcNAc-TV) is a Golgi enzyme that substitutes the trimannosyl core in the biosynthetic pathway for complex-type N-linked glycans. GlcNAc-TV activity is regulated by oncogenes frequently activated in cancer cells ( ras, src, and her2/neu ) and by activators of T lymphocytes. Overexpression of GlcNAc-TV in epithelial cells results in morphological transformation, while tumor cell mutants selected for loss of GlcNAc-TV products show diminished malignant potential in mice. In this report, we have expressed and characterized a series of N- and C-terminal deletions of GlcNAc-TV. Portions of GlcNAc-TV sequence were fused at the N-terminal domain to IgG-binding domains of staphylococcal Protein A and expressed in CHOP cells. The secreted fusion proteins were purified by IgG Sepharose affinity chromatography and assayed for enzyme activities. The peptide sequence S(213-740)of GlcNAc-TV was determined to be essential for the catalytic activity, the remaining amino acids comprising a 183 amino acid stem region, a 17 amino acid transmembrane domain and a 12 amino acid cytosolic moiety. Further deletion of 5 amino acids to produce peptide R(218-740)reduced enzyme activity by 20-fold. Similar K(m)and V(max)values for donor and acceptor were observed for peptide S(213-740), the minimal catalytic domain, and peptide Q(39-740), which also included the stem region. Truncation of five amino acids from the C-terminus also resulted in a 20-fold loss of catalytic activity. Secondary structure predictions suggest a high frequency of turns in the stem region, and more contiguous stretches of alpha-helix found in the catalytic domain. PMID- 10814702 TI - Various stages of schistosoma express Lewis(x), LacdiNAc, GalNAcbeta1-4 (Fucalpha1-3)GlcNAc and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc carbohydrate epitopes: detection with monoclonal antibodies that are characterized by enzymatically synthesized neoglycoproteins. AB - We report here that fucosylated epitopes such as Lewis(x), LacdiNAc, fucosylated LacdiNAc (LDN-F) and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF) are expressed by schistosomes throughout their life cycle. These four epitopes were enzymatically synthesized and coupled to bovine serum albumin to yield neoglycoproteins. Subsequently these neoglycoproteins were used to probe a panel of 188 monoclonal antibodies obtained from infected or immunized mice, in ELISA and surface plasmon resonance analysis. Of these antibodies, 25 recognized one of the fucosylated structures synthesized, indicating that these structures are immunogenic during infection. The MAbs identified could be subdivided in four different groups based on the recognition of either the Lewis(x)-, the LacdiNAc-, the LDN-DF-, or both the LDN-F- and LDN-DF epitope. These monoclonal antibodies were then used to investigate the localization of the fucosylated epitopes in various stages of Schistosoma mansoni using indirect immunofluorescence. Lewis(x)epitopes were mainly found in the gut and on the tegument of adult worms, on egg shells, and on the oral sucker of cercariae. The LacdiNAc epitope was expressed on the tegument of adult worms, on miracidia, and on the oral sucker of cercariae. In contrast, LDN-DF epitopes were mainly present in the excretory system of adult worms, on miracidia and on whole cercariae. These also stained positive with the LDN-F/LDN-DF epitope antibodies, while whole parenchyma reacted characteristically only with the latter antibodies. The identification of different carbohydrate structures in various stages of schistosomes may lead to a better understanding of the function of glycans in the immune response during infection. PMID- 10814703 TI - Three bovine alpha2-fucosyltransferase genes encode enzymes that preferentially transfer fucose on Galbeta1-3GalNAc acceptor substrates. AB - To investigate the synthesis of alpha2-fucosylated epitopes in the bovine species, we have characterized cDNAs from various tissues. We found three distinct alpha2-fucosyltransferase genes, named bovine fut1, fut2, and sec1 which are homologous to human FUT1, FUT2, and Sec1 genes, respectively. Their open reading frames (ORF) encode polypeptides of 360 (bovine H), 344 (bovine Se), and 368 (bovine Sec1) amino acids, respectively. These enzymes transfer fucose in alpha1,2 linkage to ganglioside GM(1)and galacto- N -biose, but not to the phenyl beta-D-galactoside, type 1 or type 2 acceptors, suggesting that their substrate specificity is different and more restricted than the other cloned mammalian alpha2-fucosyltransferases. Southern blot analyses detected four related alpha2 fucosyltransferase sequences in the bovine genome while only three have been described in other species. The supernumerary entity seems to be related to the alpha2-fucosyltransferase activity which can also use type 1 and phenyl-beta-D galactoside substrate acceptors. It was exclusively found in bovine intestinal tract. Our results show that, at least in one mammalian species, four alpha2 fucosyltransferases are present, three adding a fucose on alpha1,2 linkage on type 3/4 acceptor (Galbeta1-3GalNAc) and another able to transfer also fucose on phenyl-beta-D-galactoside and type 1 (Galbeta1-3GlcNAc) acceptors. The phylogenetic tree of the enzymes homologous to those encoded by the bovine fut1, fut2, and sec1 genes revealed two main families, one containing all the H-like proteins and the second containing all the Se-like and Sec1-like proteins. The Sec1-like family had a higher evolutionary rate than the Se-like family. PMID- 10814704 TI - Transcriptional regulation of human beta-galactoside alpha2, 6-sialyltransferase (hST6Gal I) gene during differentiation of the HL-60 cell line. AB - We have previously shown that the expression of beta-galactoside alpha2,6 sialyltransferase (hST6Gal I) mRNA decreases during HL-60 differentiation induced with dimethyl sulfoxide (DMSO) and that transcriptional regulation depends on the P3 promoter that exists 5'-upstream of exon Y (A. Taniguchi et al., FEBS Lett.,441, 191-194, 1998). The regulation of hST6Gal I may be important for the expression of sialyl-Le(x)in HL-60 cells. In the present report, we studied the transcriptional regulation of hST6Gal I gene during DMSO-induced differentiation of HL-60 cells. To elucidate the molecular basis of hST6Gal I gene expression, the genomic region containing the P3 promoter of hST6Gal I was isolated and functionally characterized. Using a luciferase assay, we identified a functional DNA portion that confers an enhancer, located at nucleotide number (nt) -317 to 174 within the P3 promoter of hST6Gal I genomic DNA. This element contains two sequences similar to Sp1 (GC-box) and one sequence similar to Oct-1 recognition motifs (octamer sequence). Site-directed mutagenesis of Sp1 and Oct-1 sites showed that two Sp1 motifs and one Oct-1 motif are essential for transcriptional activity in HL-60 cells. Enhancer activity is suppressed during HL-60 cell differentiation induced with DMSO. These results suggest that GC-box and octamer sequence may play a critical role in the transcriptional regulation of the hST6Gal I gene during HL-60 cell differentiation. PMID- 10814705 TI - In vitro characterization of anti-glucosylceramide rabbit antisera. AB - Glucosylceramides (GlcCer) are biosynthetic precursors of glycosphingolipids. They are widely distributed in biological systems where they exhibit numerous biological functions. Studies on the localization of glucosylceramides in different tissues have used biochemical methods only since specific antibodies against GlcCer were not previously available. We have characterized two commercially available rabbit antisera which were prepared against GlcCer of plant origin (1-O-(beta-D-glucopyranosyl)-N-acyl-4-hydroxysphinganine; GlcCer-3) or human origin (1-O-(beta-D-glucopyranosyl)-N-acyl-sphingosine; GlcCer-2) and claimed to be specific for GlcCer. The antisera were also able to detect specifically GlcCer species in crude lipid extracts from human epidermis after separation by thin-layer chromatography. The reagents are sensitive since both antisera reacted at dilutions higher than 1:500 with their homologous antigen in the nanogram range in thin layer immunostaining or dot-blot assays. The antisera are specific for GlcCer although they did not differentiate between GlcCer-2 and GlcCer-3 containing sphingosine or 4-hydroxysphinganine. The antisera also reacted with N-stearoyl-DL-dihydroglucocere-broside indicating that the naturally occurring structural variations in the amino alcohol moiety are not determining the specificity. No crossreactivity was observed with other mono- or diglycosylceramides (galactosylceramides, lactosyl-ceramide), free ceramides or structurally unrelated lipids (cholesterol, sphingomyelin, or phospholipids). Therefore, the glycosylmoiety seems to represent the major antigenic determinant. Finally, the antisera also proved to be useful for the immunohistochemical localization of GlcCer in human epidermis by which earlier biochemical data on the distribution of GlcCer in the various epidermal layers were confirmed. PMID- 10814706 TI - Genomic structure and promoter analysis of the human alpha1, 6-fucosyltransferase gene (FUT8). AB - GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha1,6-fucosyltransferase (alpha1,6FucT) catalyzes the transfer of a fucosyl moiety from GDP-fucose to the asparagine-linked GlcNAc residue of complex N-glycans via alpha1,6-linkage. We have cloned the genomic DNA which encodes the human alpha1,6FucT gene ( FUT8 ) and analyzed its structure. It was found that the gene consists of at least nine exons spanning more than a 50 kbp genomic region, and the coding sequence is divided into eight exons. The translation initiation codon was located at exon 2, and thus exon 1 encodes only 5'-untranslated sequences. Transcription initiation site of FUT8 was determined by 5'-rapid amplification of the cDNA end and a primer-extension analysis using the total RNA isolated from SK-OV-3 cells, which have a high level of alpha1,6FucT activity. We then characterized the FUT8 promoter region by a reporter gene assay. The luciferase reporter assay indicated that the 5'-flanking region of exon 1, which covered about 1 kbp, conferred the promoter activity in SK-OV-3 cells. This region contains potential binding sites for some transcription factors, such as bHLH, cMyb, GATA-1, as well as a TATA box, but not a CCAAT motif. 5'-Untranslated sequences found in ESTs and the cDNA for the FUT8 suggest the presence of an additional exon(s) at the upstream of the first exon identified in this study, and therefore, the transcription of the gene would be regulated by multiple promoters. PMID- 10814707 TI - Protein binding of a DRPLA family through arginine-glutamic acid dipeptide repeats is enhanced by extended polyglutamine. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is one of the hereditary neurodegenerative disorders caused by expansion of CAG/glutamine repeats. To investigate the normal function of the DRPLA gene and the pathogenic mechanism of neuron death in specific areas of the brain, we isolated and analyzed a gene that shares a notable motif with DRPLA, arginine-glutamic acid (RE) dipeptide repeats. The gene isolated, designated RERE, has an open reading frame of 1566 amino acids, of which the C-terminal portion has 67% homology to DRPLA, whereas the N terminal portion is distinctive. RERE also contains arginine-aspartic acid (RD) dipeptide repeats and putative nuclear localization signal sequences, but no polyglutamine tracts. RERE is expressed at a low level in most tissues examined. Immunoprecipitation and in vitro binding assays demonstrate that the DRPLA and RERE proteins bind each other, for which one of the RE repeats has a primary role, and extended polyglutamine enhances the binding. With engineered constructs fused with a tag, the RERE protein localized predominantly in the nucleus. Moreover, when RERE is overexpressed, the distribution of endogenous DRPLA protein alters from the diffused to the speckled pattern in the nucleus so as to co-localize with RERE. More RERE protein is recruited into nuclear aggregates of the DRPLA protein with extended polyglutamine than into those of pure polyglutamine. These results reveal a function for the DRPLA protein in the nucleus and the RE repeat in the protein-protein interaction. PMID- 10814708 TI - Decreased expression of striatal signaling genes in a mouse model of Huntington's disease. AB - To understand gene expression changes mediated by a polyglutamine repeat expansion in the human huntingtin protein, we used oligonucleotide DNA arrays to profile approximately 6000 striatal mRNAs in the R6/2 mouse, a transgenic Huntington's disease (HD) model. We found diminished levels of mRNAs encoding components of the neurotransmitter, calcium and retinoid signaling pathways at both early and late symptomatic time points (6 and 12 weeks of age). We observed similar changes in gene expression in another HD mouse model (N171-82Q). These results demonstrate that mutant huntingtin directly or indirectly reduces the expression of a distinct set of genes involved in signaling pathways known to be critical to striatal neuron function. PMID- 10814709 TI - A transcription-activating polymorphism in the ACHE promoter associated with acute sensitivity to anti-acetylcholinesterases. AB - Hypersensitivity to acetylcholinesterase inhibitors (anti-AChEs) causes severe nervous system symptoms under low dose exposure. In search of direct genetic origin(s) for this sensitivity, we studied six regions in the extended 22 kb promoter of the ACHE gene in individuals who presented adverse responses to anti AChEs and in randomly chosen controls. Two contiguous mutations, a T-->A substitution, disrupting a putative glucocorticoid response element, and a 4-bp deletion, abolishing one of two adjacent HNF3 binding sites, were identified 17 kb upstream of the transcription start site. Allele frequencies for these mutations were 0.006 and 0.012, respectively, in 333 individuals of various ethnic origins, with a strong linkage between the deletion and the biochemically neutral H322N mutation in the coding region of ACHE. Heterozygous carriers of the deletion included a proband who presented with acute hypersensitivity to the anti AChE pyridostigmine and another with unexplained excessive vomiting during a fourth pregnancy following three spontaneous abortions. Electromobility shift assays, transfection studies and measurements of AChE levels in immortalized lymphocytes as well as in peripheral blood from both carriers and non-carriers, revealed functional relevance for this mutation both in vitro and in vivo and showed it to increase AChE expression, probably by alleviating competition between the two hepatocyte nuclear factor 3 binding sites. Moreover, AChE overexpressing transgenic mice, unlike normal FVB/N mice, displayed anti-AChE hypersensitivity and failed to transcriptionally induce AChE production following exposure to anti-AChEs. Our findings point to promoter polymorphism(s) in the ACHE gene as the dominant susceptibility factor(s) for adverse responses to exposure or to treatment with anti-AChEs. PMID- 10814710 TI - Biochemical and structural analysis of missense mutations in N acetylgalactosamine-6-sulfate sulfatase causing mucopolysaccharidosis IVA phenotypes. AB - Mucopolysaccharidosis IVA (MPS IVA; OMIM#253000), a lysosomal storage disorder caused by a deficiency of N -acetylgalactosamine-6-sulfate sulfatase (GALNS), has variable clinical phenotypes. To date we have identified 65 missense mutations in the GALNS gene from MPS IVA patients, but the correlation between genotype and phenotype has remained unclear. We studied 17 missense mutations using biochemical approaches and 32 missense mutations, using structural analyses. Fifteen missense mutations and two newly engineered active site mutations (C79S, C79T) were characterized by transient expression analysis. Mutant proteins, except for C79S and C79T, were destabilized and detected as insoluble precursor forms while the C79S and C79T mutants were of a soluble mature size. Mutants found in the severe phenotype had no activity. Mutants found in the mild phenotype had a considerable residual activity (1.3-13.3% of wild-type GALNS activity). Sulfatases, including GALNS, are members of a highly conserved gene family sharing an extensive sequence homology. Thus, a tertiary structural model of human GALNS was constructed from the X-ray crystal structure of N acetylgalacto-samine-4-sulfatase and arylsulfatase A, using homology modeling, and 32 missense mutations were investigated. Consequently, we propose that there are at least three different reasons for the severe phenotype: (i) destruction of the hydrophobic core or modification of the packing; (ii) removal of a salt bridge to destabilize the entire conformation; (iii) modification of the active site. In contrast, mild mutations were mostly located on the surface of the GALNS protein. These studies shed further light on the genotype-phenotype correlation of MPS IVA and structure-function relationship in the sulfatase family. PMID- 10814711 TI - Evaluation of fine mapping strategies for a multifactorial disease locus: systematic linkage and association analysis of IDDM1 in the HLA region on chromosome 6p21. AB - The positional cloning of multifactorial disease genes is a major challenge in human genetics. We have therefore empirically tested the utility of the available polymorphic microsatellite map to locate the already identified type 1 diabetes locus IDDM1 (sibling risk/population prevalence ratio lambda(s)= 2.7) within a 14 Mb region of chromosome 6p21 linked to disease. In a two-stage approach to fine mapping, linkage was evaluated in 385 affected sib-pair families using 13 evenly spaced polymorphic microsatellite markers. The whole 14 Mb showed strong linkage. Then, each marker was analysed for evidence of allelic association, revealing evidence of disease association at one marker located within the 95% confidence interval of 1.7 cM obtained by linkage. Analysis of an additional 12 markers flanking this marker revealed a highly specific region of 570 kb associated with disease ( P = 7.5 x 10(-35)), which included the HLA class II genes, known to be the primary determinants of IDDM1. The peak of association was as close as 85 kb centromeric of the disease-predisposing class II gene HLA-DQB1. We investigated the importance of the underlying inter-marker linkage disequilibrium, marker informativity and recombination for fine mapping and demonstrate that the majority of disease association in the region can be explained by linkage disequilibrium with the class II susceptibility genes. Recombination within the major histocompatibility complex was rare and nearly absent in the class III region. We demonstrate that fine mapping of a multifactorial disease gene is possible with high accuracy even in a region with extraordinary linkage disequilibrium across distances of several Mb. The results will be applicable to association studies of disease loci with lambda(s)values <2.7 except that much larger data sets will be required. PMID- 10814712 TI - A novel protein with RNA-binding motifs interacts with ataxin-2. AB - Spinocerebellar ataxia type 2 (SCA2) is caused by expansion of a polyglutamine tract in ataxin-2, a protein of unknown function. Using the yeast two-hybrid system, we identified a novel protein, A2BP1 (ataxin-2 binding protein 1) which binds to the C-terminus of ataxin-2. Northern blot analysis showed that A2BP1 was predominantly expressed in muscle and brain. By immunocfluorescent staining, A2BP1 and ataxin-2 were both localized to the trans -Golgi network. Immunocytochemistry showed that A2BP1 was expressed in the cytoplasm of Purkinje cells and dentate neurons in a pattern similar to that seen for ataxin-2 labeling. Western blot analysis of subcellular fractions indicated enrichment of A2BP1 in the same fractions as ataxin-2. Sequence analysis of the A2BP1 cDNA revealed an RNP motif that is highly conserved among RNA-binding proteins. A2BP1 had striking homology with a human cDNA clone, P83A20, of unknown function and at least two copies of A2BP1 homologs are found in the Caenorhabditis elegans genome database. A2BP1 and related proteins appear to form a novel gene family sharing RNA-binding motifs. PMID- 10814713 TI - Evidence for a gene influencing the TG/HDL-C ratio on chromosome 7q32.3-qter: a genome-wide scan in the Framingham study. AB - Some studies show that plasma triglyceride (TG) levels are a significant independent risk factor for cardiovascular disease (CVD). TG levels are inversely correlated with high density lipoprotein cholesterol (HDL-C) levels, and their metabolism may be closely interrelated. Therefore, the TG/HDL-C ratio may be a relevant CVD risk factor. Our analysis of families in the Framingham Heart Study gave a genetic heritability estimate for log(TG) of 0.40 and for log(TG/HDL-C) of 0.49, demonstrating an important genetic component for both. A 10 cM genome-wide scan for log(TG) level and log(TG/HDL-C) was carried out for the largest 332 extended families of the Framingham Heart Study (1702 genotyped individuals). The highest multipoint variance component LOD scores obtained for both log(TG) and log(TG/HDL-C) were on chromosome 7 (at 155 cM), where the results for the two phenotypes were 1.8 and 2.5, respectively. The 7q32.3-qter region contains several candidate genes. Four other regions with multipoint LOD scores greater than one were identified on chromosome 3 [LOD score for log(TG/HDL-C) = 1.8 at 140 cM], chromosome 11 [LOD score for log(TG/HDL-C) = 1.1 at 125 cM], chromosome 16 [LOD score for log(TG) = 1.5 at 70 cM, LOD score for log(TG/HDL-C) = 1.1 at 75 cM] and chromosome 20 [LOD score for log(TG/HDL-C) = 1.7 at 35 cM, LOD score for log(TG) = 1.3 at 40 cM]. These results identify loci worthy of further study. PMID- 10814714 TI - Mutational analysis of the GPC3/GPC4 glypican gene cluster on Xq26 in patients with Simpson-Golabi-Behmel syndrome: identification of loss-of-function mutations in the GPC3 gene. AB - Simpson-Golabi-Behmel syndrome (SGBS) is an X-linked syndrome characterized by pre- and postnatal overgrowth (gigantism), which clinically resembles the autosomal Beckwith-Wiedemann syndrome (BWS). Deletions and translocations involving the glypican-3 gene ( GPC3 ) have been shown to be associated with SGBS. Occasionally, these deletions also include the glypican-4 gene ( GPC4 ). Glypicans are heparan sulfate proteoglycans which have a role in the control of cell growth and cell division. We have examined the mutational status of the GPC3 and GPC4 genes in one patient with Perlman syndrome, three patients with overgrowth without syndrome diagnosis, ten unrelated SGBS-patients and 11 BWS patients. We identified one SGBS patient with a deletion of a GPC3 exon. Six SGBS patients showed point mutations in GPC3. One frameshift, three nonsense, and one splice mutation predict a loss-of-function of the glypican-3 protein. One missense mutation, W296R, changes an amino acid that is conserved in all glypicans identified so far. A GPC3 protein that reproduces this mutation is poorly processed and fails to increase the cell surface expression of heparan sulfate, suggesting that this missense mutation is also a loss-of-function mutation. In three SGBS patients and in all non-SGBS patients, no mutations could be identified. We found three single nucleotide polymorphisms in the GPC4 gene but no evidence for loss-of-function mutations in GPC4 associated with SGBS. PMID- 10814715 TI - A full genome scan for age-related maculopathy. AB - Age-related macular degeneration or age-related maculopathy (ARM) is a major public health issue, as it is the leading cause of irreversible vision loss in the elderly in the Western world. Using three diagnostic models, we have genotyped markers in 16 plausible candidate regions and have carried out a genome wide screen for ARM susceptibility loci. A panel of 225 ARM families comprising up to 212 affected sib pairs was genotyped for 386 markers. Under our most stringent diagnostic model, the regions with the strongest evidence of linkage were on chromosome 9 near D9S301 and on 10 near D10S1230, with peak multipoint heterogeneity LOD scores (HLOD) of 1.87 and 1. 42 and peak GeneHunter-Plus non parametric LOD scores (GHP LOD) of 1. 69 and 1.83. After expanding our initial set of families to 364 ARM families with up to 329 affected sib pairs, the linkage signal on chromosome 9 vanished, while the chromosome 10 signal decreased to a GHP LOD of about 1.0, with a SimIBD P -value of 0.008 under the broadest diagnostic model with marker D10S1236. After error filtration, the GHP LOD increased to 1.27 under our most stringent model and 1.42 under our broadest model, peaking near D10S1236. This peak was seen consistently across all three diagnostic models. Our analyses also excluded up to nine different candidate regions and identified a few other regions of potential linkage, suitable for further studies. Of particular interest was the region on chromosome 5 near D5S1480, where a reasonable candidate gene, glutathione peroxidase 3, resides. PMID- 10814716 TI - KRIT1 is mutated in hyperkeratotic cutaneous capillary-venous malformation associated with cerebral capillary malformation. AB - Hyperkeratotic capillary-venous malformations (HCCVMs) are rare cutaneous lesions that occur in a small subgroup of patients with cerebral capillary malformation (CCM). CCMs cause neurological problems that range from headaches to life threatening intracranial bleeding. CCMs and HCCVMs have a similar histopathological appearance of dilated capillary-venous channels. Genetic linkage of inherited CCMs has been established to three chromosomal loci, 3q25. 2 27, 7p13-15 and 7q21-22. The first mutations were identified in the CCM1 gene (located on 7q21-22), which encodes KRIT1 protein (KREV1 interaction trapped 1), presumably a membrane-bound protein with signalling activity. Although KRIT1 is known to interact with KREV1/RAP1A, a Ras-family GTPase, the exact function of KRIT1 in the formation of cerebral capillaries and veins is poorly understood. In this study, we screened five families with CCM for mutations in the KRIT1 gene. In one of the families, CCMs co-segregated with HCCVMs. We identified a KRIT1Delta(G103)mutation in this family, suggesting that this rare form of the condition is also caused by mutations in the CCM1 gene and that KRIT1 is probably important for cutaneous vasculature. Interestingly, this deletion introduces the earliest stop codon among identified mutations, suggesting a possible correlation between the molecular alteration and the cutaneous phenotype. Another novel mutation, KRIT1(IVS2+2(T-->C)), was found in a family with only cerebral capillary-venous malformations. PMID- 10814718 TI - Rett syndrome: analysis of MECP2 and clinical characterization of 31 patients. AB - Only recently have mutations in MECP2 been found to be a cause of Rett Syndrome (RTT), a neuro-developmental disorder characterized by mental retardation, loss of expressive speech, deceleration of head growth and loss of acquired skills that almost exclusively affects females. We analysed the MECP2 gene in 31 patients diagnosed with RTT. Sequencing of the coding region and the splice sites revealed mutations in 24 females (77.40%). However, no abnormalities were detected in any of the parents that were available for investigation. Eleven mutations have not been described previously. Confirming two earlier studies, we found that most mutations are truncating and only a few of them are missense mutations. Several females carrying the same mutation display different phenotypes indicating that factors other than the type or position of mutations influence the severity of RTT. Four females with RTT variants were included in the study. Three of these presented with preserved speech while the fourth patient with congenital RTT lacked the initial period of normal development. Detection of mutations in these cases reveals that they are indeed variants of RTT. They represent the mild and the severe extremes of RTT. CONCLUSIONS: mutations in MECP2 seem to be the main cause for RTT and can be expected to be found in approximately 77% of patients that fulfil the criteria for RTT. Therefore analysis of MECP2 should be performed if RTT is suspected. Three mutation hotspots (T158M, R168X and R255X) were confirmed and a further one (R270X) newly identified. We recommend screening for these mutations before analysing the coding region. PMID- 10814717 TI - Dystrophin and utrophin influence fiber type composition and post-synaptic membrane structure. AB - The X-linked muscle wasting disease Duchenne muscular dystrophy is caused by the lack of dystrophin in muscle. Protein structure predictions, patient mutations, in vitro binding studies and transgenic and knockout mice suggest that dystrophin plays a mechanical role in skeletal muscle, linking the subsarcolemmal cytoskeleton with the extracellular matrix through its direct interaction with the dystrophin-associated protein complex (DAPC). Although a signaling role for dystrophin has been postulated, definitive data have been lacking. To identify potential non-mechanical roles of dystrophin, we tested the ability of various truncated dystrophin transgenes to prevent any of the skeletal muscle abnormalities associated with the double knockout mouse deficient for both dystrophin and the dystrophin-related protein utrophin. We show that restoration of the DAPC with Dp71 does not prevent the structural abnormalities of the post synaptic membrane or the abnormal oxidative properties of utrophin/dystrophin deficient muscle. In marked contrast, a dystrophin protein lacking the cysteine rich domain, which is unable to prevent dystrophy in the mdx mouse, is able to ameliorate these abnormalities in utrophin/dystrophin-deficient mice. These experiments provide the first direct evidence that in addition to a mechanical role and relocalization of the DAPC, dystrophin and utrophin are able to alter both structural and biochemical properties of skeletal muscle. In addition, these mice provide unique insights into skeletal muscle fiber type composition. PMID- 10814719 TI - MECP2 mutations account for most cases of typical forms of Rett syndrome. AB - Rett syndrome (RTT) is a severe progressive neurological disorder that affects almost exclusively females, with an estimated prevalence of approximately one in 10 000-15 000 female births. Most cases are sporadic, but several reports about familial recurrence support X-linked dominant inheritance with male lethality. The gene responsible for this disorder, MECP2, was recently identified by candidate gene strategy. Mutations were detected in <25% of RTT cases in this first report. To characterize the spectrum of mutations in the MECP2 gene in RTT patients, we selected 46 typical RTT patients and performed mutation screening by denaturing gradient gel electrophoresis combined with direct sequencing. We identified 30 mutations, accounting for 65% of RTT patients. They include 12 novel mutations (11 located in exon 3 and one in exon 2). Mutations, such as R270X and frameshift deletions in a (CCACC) (n) rich region, have been found with multiple recurrences. Most of the mutations were de novo, except in one family where the non-affected transmitter mother exhibited a bias of X inactivation. Although this study showed that MECP2 mutations account for most cases of typical forms of RTT (65%) and mutations in non-coding regions cannot be excluded for the remaining cases, an alternative hypothesis that takes into account the homogeneous phenotype and exclusive involvement of females, could be the implication in RTT of a putative second X-linked gene. PMID- 10814720 TI - Spectrum of Delta(7)-dehydrocholesterol reductase mutations in patients with the Smith-Lemli-Opitz (RSH) syndrome. AB - The Smith-Lemli-Opitz syndrome (SLOS; also known as the RSH syndrome) is an autosomal recessive genetic disorder, leading to characteristic multi-organ developmental abnormalities, dysmorphic facies, limb malformations and mental retardation. Mutations in the gene for Delta(7)-dehydrocholesterol reductase (Delta(7)-reductase), which catalyzes the last step in cholesterol biosynthesis, cause the disease. We screened 32 patients with SLOS, 28 from the USA and four from Sweden. Twenty-two different nucleotide changes, predicted to be disease causing mutations, were identified; 20 missense mutations, one nonsense mutation and one splice-site mutation involving the exon 9 acceptor site (IVS8 -1G-->C) were detected. All probands were heterozygous for mutations. Twelve of these mutations have not been reported previously, including missense mutations L148R, F168I, D175H, P179L, P243R, F284L, N287K, F302L, R404S, Y462H, R469P and one nonsense mutation W37X [corrected]. Coupled with previously reported mutations, these findings bring the total of different Delta(7)-reductase mutations to 36. These are distributed throughout the coding sequence of the Delta(7)-reductase gene except exons 3 and 5, with a clustering in exon 9. Three mutations account for 54% of those observed in our cohort, the splice acceptor site mutation IVS8 1G-->C (22/64 alleles, 34%), T93M (8/64, 12.5%) and V326L (5/64, 7.8%). Severity of SLOS was negatively correlated with both plasma cholesterol and relative plasma cholesterol, but not with 7-dehydrocholesterol, the immediate precursor, confirming previous observations. However, no correlation was observed between mutations and phenotype, suggesting that the degree of severity may be affected by other factors. We estimate that between 33 and 42% of the variation in the SLOS severity score is accounted for by variation in plasma cholesterol. Thus, factors other than plasma cholesterol are additionally involved in determining severity. PMID- 10814721 TI - Myopathy phenotype of transgenic mice expressing active site-mutated inactive p94 skeletal muscle-specific calpain, the gene product responsible for limb girdle muscular dystrophy type 2A. AB - A defect of the gene for p94 (calpain 3), a skeletal muscle-specific calpain, is responsible for limb girdle muscular dystrophy type 2A (LGMD2A), or 'calpainopathy', which is an autosomal recessive and progressive neuromuscular disorder. To study the relationships between the physiological functions of p94 and the etiology of LGMD2A, we created transgenic mice that express an inactive mutant of p94, in which the active site Cys129 is replaced by Ser (p94:C129S). Three lines of transgenic mice expressing p94:C129S mRNA at various levels showed significantly decreased grip strength. Sections of soleus and extensor digitorum longus (EDL) muscles of the aged transgenic mice showed increased numbers of lobulated and split fibers, respectively, which are often observed in limb girdle muscular dystrophy muscles. Centrally placed nuclei were also frequently found in the EDL muscle of the transgenic mice, whereas wild-type mice of the same age had almost none. There was more p94 protein produced in aged transgenic mice muscles and it showed significantly less autolytic degradation activity than that of wild type mice. Although no necrotic-regenerative fibers were observed, the age and p94:C129S expression dependence of the phenotypes strongly suggest that accumulation of p94:C129S protein causes these myopathy phenotypes. The p94:C129S transgenic mice could provide us with crucial information on the molecular mech anism of LGMD2A. PMID- 10814722 TI - Mutant torsinA, responsible for early-onset torsion dystonia, forms membrane inclusions in cultured neural cells. AB - Early-onset torsion dystonia is a hereditary movement disorder thought to be caused by decreased release of dopamine into the basal ganglia, without apparent neuronal degeneration. Recent cloning of the gene responsible for this disease, TOR1A (DYT1), identified the encoded protein, torsinA, as a member of the AAA+ superfamily of chaperone proteins and revealed highest levels of expression in dopaminergic neurons in human brain. Most cases of this disease are caused by a deletion of one glutamic acid residue in the C-terminal region of the protein. Antibodies generated against torsinA revealed expression of a predominant immunoreactive protein species similar to the predicted size of 37.8 kDa in neural, glial and fibroblastic lines by western blot analysis. This protein is N glycosylated with high mannose content and not, apparently, phosphoryl-ated. Overexpression of torsinA in mouse neural CAD cells followed by immunocytochemistry, revealed a dramatically different pattern of distribution for wild-type and mutant forms of the protein. The wild-type protein was found throughout the cytoplasm and neurites with a high degree of co-localization with the endoplasmic reticulum (ER) marker, protein disulfide isomerase. In contrast, the mutant protein accumulated in multiple, large inclusions in the cytoplasm around the nucleus. These inclusions were composed of membrane whorls, apparently derived from the ER. If disrupted processing of the mutant protein leads to its accumulation in multilayer membranous structures in vivo, these may interfere with membrane trafficking in neurons. PMID- 10814723 TI - Disruption of two novel genes by a translocation co-segregating with schizophrenia. AB - A balanced (1;11)(q42.1;q14.3) translocation segregates with schizophrenia and related psychiatric disorders in a large Scottish family (maximum LOD = 6.0). We hypothesize that the translocation is the causative event and that it directly disrupts gene function. We previously reported a dearth of genes in the breakpoint region of chromosome 11 and it is therefore unlikely that the expression of any genes on this chromosome has been affected by the translocation. By contrast, the corresponding region on chromosome 1 is gene dense and, not one, but two novel genes are directly disrupted by the translocation. These genes have been provisionally named Disrupted-In Schizophrenia 1 and 2 ( DISC1 and DISC2 ). DISC1 encodes a large protein with no significant sequence homology to other known proteins. It is predicted to consist of a globular N-terminal domain(s) and helical C-terminal domain which has the potential to form a coiled-coil by interaction with another, as yet, unidentified protein(s). Similar structures are thought to be present in a variety of unrelated proteins that are known to function in the nervous system. The putative structure of the protein encoded by DISC1 is therefore compatible with a role in the nervous system. DISC2 apparently specifies a non-coding RNA molecule that is antisense to DISC1, an arrangement that has been observed at other loci where it is thought that the antisense RNA is involved in regulating expression of the sense gene. Altogether, these observations indicate that DISC1 and DISC2 should be considered formal candidate genes for susceptibility to psychiatric illness. PMID- 10814724 TI - Linkage and linkage disequilibrium in chromosome band 1p36 in American Chaldeans with inflammatory bowel disease. AB - The idiopathic inflammatory bowel diseases (IBDs), consisting of Crohn's disease and ulcerative colitis, are complex genetic disorders involving chronic inflammation of the intestines. Multiple genetic loci have been implicated through genome-wide searches, but refinement of localization sufficient to undertake positional cloning efforts has been problematic. This difficulty can be obviated through identification of ancestrally shared regions in genetic isolates, such as the Chaldean population, a Roman Catholic group from Iraq. We analyzed four multiply affected American Chaldean families with inflammatory bowel disease not known to be related. We observed evidence for linkage and linkage disequilibrium in precisely the same region of chromosome band 1p36 reported previously in an outbred population. Maximal evidence for linkage was observed near D1S1597 by multipoint analysis (MLOD = 3.01, P = 6.1 x 10(-5)). A shared haplotype (D1S507 to D1S1628) was observed over 27 cM between two families. There was homozygous sharing of a 5 cM portion of that haplotype in one family and over a <1 cM region in the second family. Homozygous sharing of this haplotype near D1S2697 and D1S3669 was observed in one individual in a third multiply affected family, with heterozygous sharing in a fourth family. Linkage in outbred families as well as in this genetic isolate indicates that a pathophysiologically crucial IBD susceptibility gene is located in 1p36. These findings provide a unique opportunity to refine the localization and identify a major susceptibility gene for a complex genetic disorder. PMID- 10814725 TI - Barhl1, a gene belonging to a new subfamily of mammalian homeobox genes, is expressed in migrating neurons of the CNS. AB - The BarH1 and BarH2 ( Bar ) Drosophila genes are homeobox-containing genes, which are required for the fate determination of external sensory organs in the fly. By means of a bioinformatic approach, we have identified in mouse and human two homeobox genes highly related to the Bar Drosophila genes, Barhl1 and Barhl2. While Barhl1 represents a novel gene, Barhl2 turned out to correspond to the mBH1 cDNA recently described in rat. We isolated and sequenced the full-length mouse Barhl1 and mapped both the human BARHL1 and BARHL2 genes to chromosomes 9q34 and 1p22, respectively. Detailed analysis of the murine Barhl1 expression pattern by in situ hybridization revealed that this transcript is exclusively expressed in restricted domains of the developing CNS, which suggests that this gene, similar to its Drosophila counterparts BarH1 and BarH2, may play a crucial role in cell fate determination of neural structures. In particular, Barhl1 showed specific domains of expression in the diencephalon and in the rhombencephalon where it was found to be expressed in migrating cells giving rise to the cerebellar external granular layer and to specific populations of dorsal sensory interneurons of the spinal cord. Thus, Barhl1 function may be required for the generation of these specific subtypes of neuronal progenitors. Furthermore, the mapping assignment and the expression pattern make BARHL1 an attractive positional candidate gene for a form of Joubert syndrome, a rare developmental anomaly of the cerebellum in humans. PMID- 10814726 TI - Identification of mutations in the gene encoding lamins A/C in autosomal dominant limb girdle muscular dystrophy with atrioventricular conduction disturbances (LGMD1B). AB - LGMD1B is an autosomal dominantly inherited, slowly progressive limb girdle muscular dystrophy, with age-related atrioventricular cardiac conduction disturbances and the absence of early contractures. The disease has been linked to chromosome 1q11-q21. Within this locus another muscular dystrophy, the autosomal dominant form of Emery-Dreifuss muscular dystrophy (AD-EDMD) has recently been mapped and the corresponding gene identified. AD-ADMD is characterized by early contractures of elbows and Achilles tendons and a humero peroneal distribution of weakness combined with a cardiomyopathy with conduction defects. The disease gene of AD-EDMD is LMNA which encodes lamins A/C, two proteins of the nuclear envelope. In order to identify whether or not LGMD1B and AD-EDMD are allelic disorders, we carried out a search for mutations in the LMNA gene in patients with LGMD1B. For this, PCR/SSCP/sequencing screening was carried out for the 12 exons of LMNA on DNA samples of individuals from three LGMD1B families that were linked to chromo-some 1q11-q21. Mutations were identified in all three LGMD1B families: a missense mutation, a deletion of a codon and a splice donor site mutation, respectively. The three mutations were identified in all affected members of the corresponding families and were absent in 100 unrelated control subjects. The present identification of mutations in the LMNA gene in LGMD1B demonstrates that LGMD1B and AD-EDMD are allelic disorders. Further analysis of phenotype-genotype relationship will help to clarify the variability of the phenotype observed in these two muscular dystrophies. PMID- 10814727 TI - Physiological characterization of layer III non-pyramidal neurons in piriform (olfactory) cortex of rat. AB - We performed whole-cell recordings of layer III non-pyramidal neurons in the piriform cortex of Sprague-Dawley rats. For comparison purposes, recordings were made from deep pyramidal cells, which are also present in layer III. These two cell types could be distinguished both anatomically and physiologically. Anatomically, the layer III non-pyramidal neuron displayed smooth beady dendrites, while deep pyramidal cells showed thicker dendrites with spines. The dendrites of the layer III non-pyramidal neuron also tended to be restricted to layer III while deep pyramidal cells had long apical dendrites that spanned layers I and II. Although the resting membrane potentials of both cell types were very similar, significant differences were noted in other physiological measures. Layer III non-pyramidal neurons typically displayed higher input resistances, faster time constants, smaller spike amplitudes, shorter spike widths, and higher spike thresholds. In addition, layer III non-pyramidal neurons were able to spike at much higher rates when stimulated with the same level of threshold normalized current injection. The most dramatic differences in physiology were seen in the pattern of spiking in response to increasing levels of positive constant current pulses. Layer III non-pyramidal neurons showed qualitatively different responses at low and high levels of stimulation. At low levels, spikes occurred with long latency and the firing frequency increased throughout the duration of the current pulse. At high levels, non-pyramidal neurons started spiking with short latency, followed by a decrease in firing frequency, which in turn was followed by an increase in firing frequency. Deep pyramidal neurons differed dramatically from this pattern, displaying a qualitatively similar response at all levels of current injection. This response was characterized by short latency spikes and spike adaptation for the duration of the current pulse. PMID- 10814728 TI - Glucocorticosteroids stimulate the activity of the promoters of peripheral myelin protein-22 and protein zero genes in Schwann cells. AB - To better understand the mechanism by which glucocorticosteroids (GLUC) could enhance myelination in the PNS, cultured rat Schwann cells were transiently transfected with reporter constructs in which luciferase expression was controlled by the promoter region of either the peripheral myelin protein-22 (PMP22) or the protein zero (P(0)) genes. GLUC stimulated the activity of the P(0) promoter and the PMP22 promoters 1 and 2. The effect of GLUC was specific as estradiol and testosterone did not activate the promoters. The antagonist RU486 did not abolish the effect of GLUC, but instead stimulated promoter activities by itself. In the mammary carcinoma cell line 34i, which expresses GLUC receptors, GLUC did not stimulate the P(0) and PMP22 promoters while the promoter of the mouse mammary tumor virus was strongly activated. Thus, the activation by GLUC of the promoter activities of two peripheral myelin protein genes is Schwann cell specific. PMID- 10814729 TI - Antinociception produced by mu opioid receptor activation in the amygdala is partly dependent on activation of mu opioid and neurotensin receptors in the ventral periaqueductal gray. AB - Exposure to stressful or fear-inducing environmental stimuli activates descending antinociceptive systems resulting in a decreased pain response to peripheral noxious stimuli. Stimulating mu opioid receptors in the basolateral nucleus of the amygdala (BLA) in anesthetized rats produces antinociception that is similar to environmentally induced antinociception in awake rats. Recent evidence suggests that both forms of antinociception are mediated via projections from the amygdala to the ventral periaqueductal gray (PAG). In the present study, we examined the types of neurochemicals released in the ventral PAG that may be important in the expression of antinociception produced by amygdala stimulation in anesthetized rats. Microinjection of a mu opioid receptor agonist into the BLA resulted in a time dependent increase in tail flick latency that was attenuated by preadministration of a mu opioid receptor or a neurotensin receptor antagonist into the ventral PAG. Microinjection of a delta(2) opioid receptor antagonist or an NMDA receptor antagonist into the ventral PAG was ineffective. These findings suggest that amygdala stimulation produces antinociception that is mediated in part by opioid and neurotensin release within the ventral PAG. PMID- 10814730 TI - Neuronal expression of Fos protein in the hypothalamus of rats with heart failure. AB - We sought to identify the areas that have altered neuronal activity within the hypothalamus of rats with heart failure (HF) by mapping neuronal staining of c Fos protein (Fos) 6-8 weeks following coronary artery ligation (HF group; n=17) or sham surgery (sham-operated control group, n=15). Fos-like immunoreactivity was observed in the paraventricular nucleus (PVN), supraoptic nucleus (SON), median preoptic nucleus (MnPO), anterior hypothalamus (AH) and posterior hypothalamus (PH) using a standard ABC immunocytochemical protocol. The rats in the HF group displayed infarcts averaging 34+/-2% of the outer circumference and 41+/-1% of the inner circumference of the left ventricular wall. Sham-operated control rats had no observable damage to the myocardium. Rats with chronic heart failure (n=5) but no manipulation (no surgery) had a similar number of Fos staining cells in PVN SON, MnPO, AH and PH compared to sham-operated rats. Acute surgery for isolation of vagus nerves and anesthesia for 90 min increased the number of Fos positive cells in PVN, SON and MnPO of both sham-operated rats and rats with HF. Furthermore, rats with heart failure (n=5) had significantly higher number of Fos-staining cells in PVN (four times), SON (4.5 times) and MnPO (1.5 times) compared to sham-operated rats after acute surgery for isolation of the vagus. The number of Fos-staining cells remained unaltered in AH and PH in both groups of rats. However, in a third series of experiments vagotomy reduced the number of Fos-staining cells in the PVN, SON or MnPO of rats with HF (n=5) to those observed in sham-operated vagotomized rats. This study shows that: (1) there is augmented neuronal activity as indicated by increased number of Fos staining neurons in the PVN, SON and MnPO due to acute surgical stress in rats with HF, and (2) vagal afferents are responsible for the increased neuronal activity in PVN, SON and MnPO of rats with HF during acute surgical stress. These data support the conclusion that vasopressin producing neurons and autonomic areas within the hypothalamus influenced by vagal afferents are activated during HF and are sensitive to 'acute surgical stress' and may contribute to the elevated levels of vasopressin and sympatho-excitation commonly observed in heart failure. PMID- 10814731 TI - A novel neurotensin peptide analog given extracranially decreases food intake and weight in rodents. AB - Neurotensin decreases food intake in the rat when injected into the cerebral ventricles. We tested the effect of a novel neurotensin analog (NT69L), injected intra-peritoneally (i.p.), on weight gain and food intake in rats. Sprague-Dawley rats (270 g) were injected i. p. with either saline or NT69L at 0.001 or 0.010 mg/kg. In further experiments, larger rats at a more steady state on the growth curve (400 g) were injected with either saline or 0.010 or 1 mg/kg NT69L. Food intake, water consumption and body weight were recorded daily. Weight gain was significantly reduced in the smaller rats injected with 0.001 or 0.010 mg/kg, showing only a 8.5 and 9.0% increase in original weight, respectively, as compared to a 29% increase for the controls. The larger rats injected with 1 mg/kg, had a significant reduction in body weight with a 3.0% decrease in original body weight as compared to a 2.4% increase for the controls. Food intake was significantly reduced suggesting that the weight loss observed after injection of NT69L was attributable in part to a reduction in food intake. The genetically obese Zucker rats injected with NT69L (1 mg/kg) had a significant reduction in weight gain and food intake. NT69L significantly increased blood glucose and corticosterone levels and decreased TSH and T4 in Sprague-Dawley and Zucker rats, an effect that was only transitory. NT69L also caused a decrease in norepinephrine in both the hypothalamus and nucleus accumbens, and an increase in dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and serotonin. In this study, NT69L exhibited a consistent and dramatic effect on body weight and food intake in Sprague-Dawley and obese Zucker rats, and enabled us to study the role that NT plays in weight control and the functional interactions of NT with brain amines, and metabolic and endocrinological parameters. PMID- 10814732 TI - Postnatal development of a GABA deficit and disturbance of neural functions in mice lacking GAD65. AB - The 65-kDa isoform of glutamic acid decarboxylase (GAD65) is believed to play an essential role for GABA synthesis in the central nervous system. Using mice with targeted disruption of the GAD65 gene (GAD65(-/-) mice) we investigated the contribution of GAD65 to GABA synthesis in different brain areas during postnatal development and in adulthood. In the amygdala, hypothalamus and parietal cortex of GAD65(+/+) mice an increase of GABA levels was observed during postnatal development, most prominently between the first and second month after birth. This increase appeared to be dependent on GAD65, as it was delayed by 2 months in GAD65(+/-) mice and was not observed in GAD65(-/-) mice. Likely as a consequence of their GABA deficit, adult GAD65(-/-) mice showed a largely abnormal neural activity with frequent paroxysmal discharges and spontaneous seizures. They furthermore displayed increased anxiety-like behaviour in a light/dark avoidance test and reduced intermale aggression, as well as a reduced forced-swimming induced immobility indicative of an antidepressant-like behavioural change. Adult GAD65(+/-) mice did not show behavioural disturbances except for a reduced aggressive behaviour that was comparable to that in GAD65(-/-) mice. We conclude that GAD65-mediated GABA synthesis may be crucially involved in control of emotional behaviour and indispensable for a tonic inhibition that prevents the development of hyperexcitability in the maturating central nervous system. Aggressive, and possibly other social behaviour may be especially prone to regulation through GAD65-mediated GABA synthesis. PMID- 10814733 TI - Transfection of C6 glioma cells with glia maturation factor upregulates brain derived neurotrophic factor and nerve growth factor: trophic effects and protection against ethanol toxicity in cerebellar granule cells. AB - Glial cells play active roles in neuronal survival, as well as neuroprotection against toxic insult. Recent studies suggest that the brain protein glia maturation factor (GMF) is involved in intracellular signaling in glia. This study investigated whether or not GMF plays a role in the survival-promoting and neuroprotective functions of glia. C6 glioma cells were transfected in vitro with GMF utilizing an adenovirus vector. The transfected cells overexpressed GMF intracellularly, but did not secrete the protein. The conditioned medium (CM) was obtained from the GMF-transfected cells (CM-GMF) and tested on primary neuronal cultures, consisting of cerebellar granule cells (CGC). The CGC cultures were utilized because these cultures have a background level of cell death, and the survival-promoting, i.e. neurotrophic effect, of the CM could be tested. In addition, since CGC cultures are ethanol-sensitive (ethanol enhances neuronal death), the neuroprotective effect of the CM against ethanol-induced cell death was tested also. We demonstrated that the CM-GMF had an enhanced neurotrophic effect as well as an increased neuroprotective effect against ethanol-induced cell death compared to control CM obtained from untransfected C6 cells (CM-Mock) or CM obtained from cells transfected with an unrelated gene (CM-LacZ). Because neurotrophins have trophic and protective effects, we investigated whether GMF transfection upregulated the expression of neurotrophins in C6 cells. RT-PCR verified that GMF-transfected C6 cells had increased mRNA levels for BDNF and NGF. Immunoblotting corroborated the RT-PCR results and indicated that CM-GMF contained greater concentrations of BDNF and NGF protein compared to CM-Mock and CM-LacZ. A soluble TrkB-IgG fusion protein, which selectively binds BDNF and prevents its binding to the neuronal TrkB receptor, eliminated the neurotrophic effect of CM-GMF; whereas anti-NGF antibody was ineffective in preventing this effect, suggesting that the neurotrophic effect was due to BDNF. On the other hand, both the TrkB-IgG fusion protein and anti-NGF reduced neuroprotection, suggesting that BDNF and NGF both contribute to the neuroprotective effect of CM GMF. In conclusion, GMF upregulates the expression of BDNF and NGF in C6 cells, and these factors exert neurotrophic and neuroprotective functions on primary neurons. PMID- 10814734 TI - Spatial learning-induced increase in the argyrophilic nucleolar organizer region of dorsolateral telencephalic neurons in goldfish. AB - Spatial learning and memory related morphological changes in the argyrophilic nucleolar organizer region (AgNOR) of telencephalic neurons in goldfish were quantitatively evaluated by means of AgNOR neurohistochemical stain. The AgNORs and nuclei of nerve cells of two different telencephalic regions of goldfish trained in a spatial task or submitted to a similar non-contingent behavioral procedure (control group) were morphometrically evaluated. Results show that the area of AgNORs in goldfish dorsolateral telencephalic neurons increased significantly in the spatial learning group but not in control group. This effect seems to be highly specific as it did not appear in the dorsolateral area of the control group neither in the dorsomedial area of both groups. As the size of AgNORs in the nerve cell nuclei reflect the level of transcriptive activity, these morphological changes could be revealing increased protein synthesis in goldfish dorsolateral telencephalic neurons related with learning and memory. These findings could contribute to determining the subregions of the teleost telencephalon implicated in spatial learning and could indicate that the AgNOR staining technique would be a useful tool in assesing learning and memory related neuronal activity. PMID- 10814735 TI - Biochemical and electrophysiological evidence that RO 60-0175 inhibits mesolimbic dopaminergic function through serotonin(2C) receptors. AB - In vivo microdialysis and electrophysiological techniques were used to elucidate the role of the 5-HT(2) receptor family on the control of mesolimbic dopaminergic system exerted by serotonin (5-HT). Administration of RO 60-0175 (1 mg/kg, i.p.), a selective 5-HT(2C) receptor agonist, significantly decreased dopamine (DA) release by 26+/-4% (below baseline) 60 min after injection. Moreover, RO 60-0175 (80-320 microg/kg, i.v.) dose-dependently decreased the basal firing rate of DA neurons in the ventral tegmental area (VTA), reaching its maximal inhibitory effect (53.9+/-15%, below baseline) after the dose of 320 microg/kg. The selective 5-HT(2C) receptor antagonist SB 242084 completely blocked the inhibitory action of RO 60-0175 on accumbal DA release and on the firing rate of VTA DA cells. On the contrary, both (+/-)-DOI, a mixed 5-HT(2A/2C) receptor agonist, and the selective 5-HT(2B) agonist BW 723C86, did not affect either DA release in the nucleus accumbens or the firing rate of VTA DA cells. Taken together, these data confirm that central 5-HT system exerts an inhibitory control on the mesolimbic DA system and that 5-HT(2C) receptors are involved in this effect. PMID- 10814736 TI - Evaluation of the role for prolactin-releasing peptide in prolactin secretion induced by ether stress and suckling in the rat: comparison with vasoactive intestinal peptide. AB - Prolactin (PRL)-releasing peptide (PrRP) is a recently discovered hypothalamic peptide possessing a specific stimulatory action on PRL secretion. In this study, we examined whether PrRP plays a role in mediating ether stress- and suckling induced PRL secretion in rats through administering anti-PrRP antisera intracerebroventricularly. For comparison, we also tested the effect of anti vasoactive intestinal peptide (VIP) antisera on the hormonal responses, since VIP is another candidate for a physiological PRL-releasing factor. The immunoneutralization of VIP, but not of PrRP, led to a significant suppression of PRL responses to both ether and suckling. These results suggest that PrRP may not play a significant role, or at least play a much weaker role than VIP, in mediating PRL release induced by ether stress and suckling in the rat. PMID- 10814737 TI - A significant role of leukemia inhibitory factor in the brain and periphery in endotoxin stimulation of adrenocorticotropin secretion in the rat. AB - Leukemia inhibitory factor (LIF) is considered as another important cytokine regulating the activity of the hypothalamo-pituitary-adrenal axis. In this study, we examined the effects of intravenous (iv) and intracerebroventricular (icv) administrations of anti-LIF antibody on plasma adrenocorticotropin (ACTH) responses induced by intraperitoneal administration of lipopolysaccharide (LPS, 250 microg/kg) in male rats. Fifteen minutes before the LPS injection, anti-rat LIF antibody or control serum was given iv or icv. The antibody was administered at two different concentrations, i.e. undiluted and five-times diluted. Irrespective of the route of administration, the anti-LIF antibody partially but significantly suppressed ACTH responses to LPS, and its suppressive effect was similar between its two different concentrations. These results indicate that the anti-LIF antibody already exerted its maximal effects at its diluted preparation, and hence that the role of LIF in LPS-stimulated ACTH secretion is essentially partial. This is the first study to demonstrate in vivo that LIF in both the brain and general circulation plays a significant role in mediating endotoxin stimulated ACTH secretion in the rat. PMID- 10814738 TI - Complement receptor 3-like immunoreactivity in the light green cells and the canopy cells of the pond snail, Lymnaea stagnalis. AB - We observed CR3-like immunoreactivity in the central nervous system (CNS) and its surrounding peripheral nerves of the pond snail, Lymnaea stagnalis. In the CNS of L. stagnalis, the immunoreactivity presenting meshwork-like structure was detected in some neurosecretory cells, which are the light green cells (LGCs) and the canopy cells (CCs), both controlling the body growth. The immunoreactivity was also observed along the edges of median lip nerves. The immunoreactive regions in the median lip nerves appeared to form the axonal plates, from which the LGCs and the CCs release molluscan insulin-related peptides (MIPs) into the blood. By contrast, no immunoreactivity was detected in other neurosecretory cells or their release sites, for example the caudodorsal cells and the cerebral commissure, which release ovulation hormones. The present findings, therefore, suggested that CR3 expresses only in the neurosecretory cells releasing MIPs in L. stagnalis. PMID- 10814739 TI - Rhythmic midbrain-evoked vocalization is inhibited by vasoactive intestinal polypeptide in the teleost Porichthys notatus. AB - Vasoactive intestinal polypeptide (VIP) is distributed in vocal midbrain areas of multiple vertebrate taxa, suggesting that VIP may modulate midbrain-evoked vocalization. To test this hypothesis, neurophysiological experiments were conducted in the teleost Porichthys notatus which generates vocalizations in mating and agonistic contexts. Electrical stimulation of the paralemniscal midbrain and local delivery of VIP were conducted in conjunction with occipital nerve recordings that reflect the patterned output of hindbrain vocal circuitry. Consistent with our hypothesis, VIP significantly reduced the duration and number of rhythmic vocal-motor bursts obtained in a dose-dependent manner; vocalization latency was concomitantly increased. These results provide the first evidence for VIP modulation of midbrain vocal function. PMID- 10814740 TI - Nitric oxide involvement in the trigeminal hyperalgesia in diabetic rats. AB - Trigeminal hyperalgesia frequently appears in diabetic neuralgia altering the transmission of orofacial sensory information. This study was designed to explore the effects of trigeminal hyperalgesia in streptozotocin-induced diabetes monitoring the expression of nitric oxide synthase in the trigeminal ganglion cells. The threshold to heat noxious stimuli decreased in diabetic animals. The number of NADPH-diaphorase (NADPH-d)-positive neurons significantly decreased in the diabetic rats compared with controls. Insulin treatment prevented the decreased nociceptive threshold and reduction of the number of NADPH-d-positive neurons. These findings point out that there is a relationship between the trigeminal nociceptive perception and NADPH-d neuronal expression suggesting that NO may play a role in the pathogenesis of trigeminal sensory neuropathy. PMID- 10814741 TI - Casein kinase 1 delta mRNA is upregulated in Alzheimer disease brain. AB - The casein kinase-1 (Ck1) family are serine/threonine specific protein kinases. They are highly associated with Alzheimer disease (AD) brain-derived tau filaments and granulovacuolar bodies. Recently we have demonstrated that one family member, Ckidelta, colocalizes with tau containing neurofibrillary tangles (NFTs) and other tau deposits in a number of neurodegenerative diseases. Here we show that the association in AD is accompanied by a sharp upregulation of Ckidelta mRNA in brain but not in peripheral organs. The degree of upregulation in AD brain is correlated with the degree of regional pathology. There was a 24.4 fold increase of Ckidelta mRNA in AD hippocampus compared with control, 8.04-fold in the amygdala, 7.45 in the entorhinal cortex and 7.30-fold in the midtemporal gyrus. These are areas with a high burden of NFTs, neuropil threads and dystrophic neurites. In areas almost devoid of this tau pathology, such as the caudate nucleus, occipital cortex and cerebellum, the increases in AD compared to control brain were only 2.21-, 1.89- and 1.87-fold, respectively. Western blot analysis showed that the upregulation of Ckidelta mRNA was paralleled by an upregulation of Ckidelta protein. These data establish that the association of Ckidelta with the tau pathology of AD is reflective of an increase in gene transcription. Since Alzheimer-like phosphoepitopes of tau can be generated by Ck1, the Ckidelta isoform may play an important role in this fundamental aspect of AD pathology. PMID- 10814742 TI - Expression of connexin36 in the adult and developing rat brain. AB - The distribution of connexin36 (Cx36) in the adult rat brain and retina has been analysed at the protein (immunofluorescence) and mRNA (in situ hybridization) level. Cx36 immunoreactivity, consisting primarily of round or elongated puncta, is highly enriched in specific brain regions (inferior olive and the olfactory bulb), in the retina, in the anterior pituitary and in the pineal gland, in agreement with the high levels of Cx36 mRNA in the same regions. A lower density of immunoreactive puncta can be observed in several brain regions, where only scattered subpopulations of cells express Cx36 mRNA. By combining in situ hybridization for Cx36 mRNA with immunohistochemistry for a general neuronal marker (NeuN), we found that neuronal cells are responsible for the expression of Cx36 mRNA in inferior olive, cerebellum, striatum, hippocampus and cerebral cortex. Cx36 mRNA was also demonstrated in parvalbumin-containing GABAergic interneurons of cerebral cortex, striatum, hippocampus and cerebellar cortex. Analysis of developing brain further revealed that Cx36 reaches a peak of expression in the first two weeks of postnatal life, and decreases sharply during the third week. Moreover, in these early stages of postnatal development Cx36 is detectable in neuronal populations that are devoid of Cx36 mRNA at the adult stage. The developmental changes of Cx36 expression suggest a participation of this connexin in the extensive interneuronal coupling which takes place in several regions of the early postnatal brain. PMID- 10814743 TI - Effects of three neurochemical stimuli on delayed feeding and energy metabolism. AB - Infusions of norepinephrine (NE), the gamma-aminobutyric acid agonist, muscimol (MUS), or neuropeptide Y (NPY) into the paraventricular nucleus (PVN) of the hypothalamus all increase food intake. Such feeding may be due to direct activation of behavioral processes driving ingestion and/or to alterations in nutrient metabolism that feeding serves to normalize. To examine these possibilities, male Sprague-Dawley rats received PVN infusions of vehicle, 20 nmol NE, 1 nmol MUS or 100 pmol NPY at dark onset, then food intake was measured under three feeding conditions: (1) 1 and 2 h immediately after injections, (2) 1 h after a 1 h delay between injections and access to food, and (3) 1 h after a 1 h feeding delay, but with injections occurring just before presenting food. Measures of energy expenditure (EE) and respiratory quotients (RQs) in the absence of food were made over 2 h in parallel experiments. Results confirmed that NE, MUS and NPY all increased dark-onset feeding, but only NPY increased intake above control levels after a 1 h feeding delay. No neurochemically-induced changes in EE were observed, nor were there changes in RQs after NE or MUS. However, NPY reliably enhanced RQs from 30 to 120 min of testing. Our findings imply that NE and MUS initiate relatively immediate, short-term feeding that is not associated with changes in nutrient metabolism and does not summate with cues stimulated by delayed access to food. NPY initiates more protracted feeding temporally linked to enhanced carbohydrate metabolism. This may indicate that part of NPY's feeding stimulatory effects are secondary to physiological processes driving ingestion. PMID- 10814744 TI - Introduction PMID- 10814745 TI - Neoral use in the liver transplant recipient. AB - This review and critical analysis of current trends of immunosuppression management in liver transplantation provides evidence and support for the continued role of Neoral as an indispensable part of immunosuppressive protocols. CyA formulation influences clinical outcomes such as acute rejection; this is confirmed by several multicenter studies. The CyA microemulsion formulation provides more reliable and effective absorption characteristics, and it may provide more rapid and complete immunosuppression in the de novo patient than IV administration. An advanced TDM strategy, particularly a 2-hour post-dose blood level (C-2), can improve the effectiveness and safety of immunosuppression in de novo liver transplant patients. There are potential risks resulting from CyA withdrawal strategies, as there is no evidence supporting this strategy. Neoral is an indispensable part of combination protocols in liver transplantation. PMID- 10814746 TI - Neoral use in the renal transplant recipient. AB - This review and critical analysis of current trends of immunosuppression management in pediatric transplantation provides evidence and support for the continued role of Neoral as an indispensable part of immunosuppressive protocols. CyA formulation influences clinical outcomes such as acute rejection, as confirmed by several multicenter studies. The CyA microemulsion formulation reduces pharmacokinetic variability and its consequent impact on outcomes over the long term. An advanced TDM strategy can improve the effectiveness and safety of immunosuppression in both de novo and maintenance renal transplant patients. There are potential risks resulting from CyA withdrawal strategies. Neoral is an indispensable part of combination protocols in renal transplantation. PMID- 10814747 TI - Neoral use in the pediatric transplant recipient. AB - This review and critical analysis of current trends of immunosuppression management in pediatric transplantation provides evidence and support for the continued role of Neoral as an indispensable part of immunosuppressive protocols. CyA formulation influences clinical outcomes such as acute rejection, as confirmed by two studies. The CyA microemulsion formulation provides more reliable and effective absorption. An advanced TDM strategy to determine CyA bioavailability can improve the effectiveness and safety of immunosuppression in de novo liver transplant patients. especially in the younger de novo patient. Neoral is an indispensable part of combination protocols in pediatric transplantation. PMID- 10814748 TI - Neoral use in the cardiac transplant recipient. AB - CyA is the core immunosuppressant of choice for the majority of transplant patients. The introduction of Neoral, a new microemulsion formulation of CyA. and more recently a range of adjunctive immunosuppressants have further enhanced the efficacy and tolerability of CyA-based immunosuppression. In the first year following transplantation the major causes of morbidity and death are graft failure, acute rejection, and systemic infection. Patients with deteriorated pulmonary circulation before transplantation are at increased risk of early postoperative death. Risk factors for early acute rejection include female donor sex, young donor age, and multiple HLA-DR mismatches. The principal cause of death in the long term is graft vasculopathy which accounted for 40% of all deaths. Risk factors that have been hypothesized to play a role in the pathogenesis of graft vasculopathy include hyperlipidemia, recipient age and gender, donor age, the number of HLA AB and DR mismatches, and CMV infection. Strategies proposed to reduce the risk of graft vasculopathy include aggressive use of lipid-lowering agents, avoidance of low CyA doses, and the use of adjunctive rapamycin or RAD therapy. Rejection surveillance therefore relies on routine serial endomyocardial biopsy. Recent research suggests that a more accurate assessment of the state of the graft can be obtained by considering the results across a number of biopsy samples obtained from different parts of the heart, rather than basing clinical judgment on the worst single result obtained. New molecular markers such as granzyme A mRNA are likely to improve the power of histology to diagnose and predict rejection. Neoral pharmacokinetics give greater bioavailability and less intrapatient variability than Sandimmune. In the keynote OLN 351 study comparing Neoral with Sandimmune in de novo heart transplant recipients, fewer Neoral patients needed antilymphocyte therapy to treat rejection, fewer female patients had rejection episodes in the Neoral group, the tolerability of the two formulations was equivalent, and there was a lower incidence of infections in the Neoral group. The clinical impact of Neoral in comparison with Sandimmune in de novo heart transplant patients has been investigated in a number of additional trials, including long-term studies, which have confirmed that Neoral is associated with: Lower CyA doses than Sandimmune. Equal or greater antirejection efficacy than Sandimmune. Comparable tolerability to Sandimmune. During the administration of intravenous CyA as an induction therapy in the days immediately following transplantation, there is evidence to suggest that a 6-hour infusion given twice daily, which mimics the pharmacokinetic profile of oral dosing, may be clinically more effective than a continuous 24-hour infusion. Milligram-for-milligram dose conversion from Sandimmune to Neoral is feasible. Following conversion, a reduction in the CyA dose may be required in the majority of patients to maintain target levels. In pediatric patients, the rate of elimination of CyA is greater and bioavailability increases with increasing age. Younger patients (less than 8 years of age) may be managed more effectively with a 3-times-daily, rather than a twice-daily dosing schedule. A number of studies have compared the clinical effects of Sandimmune and Neoral in maintenance therapy for cardiac transplant patients. As with de novo patients, these studies have found the new formulation of CyA to be associated with lower rates of acute rejection, lower therapeutic doses, and comparable tolerability. Milligram-to-milligram conversion from the old to the new CyA formulation is generally well tolerated, although in a minority of patients there is a significant increase in CyA levels. These may be associated with a transient increase in side effects which resolve on dose reduction. There is a dose-sparing effect with Neoral. Routine monitoring of both CyA and serum creatinine levels are adv PMID- 10814749 TI - Neoral absorption profiling: an evolution in effectiveness. PMID- 10814750 TI - Pharmacokinetic validation of neoral absorption profiling. PMID- 10814751 TI - Color, form and luminance capture attention in visual search. AB - Extant models of visual attention predict that a salient element should produce a bottom-up activation leading to a stimulus-driven attentional capture (e.g. Cave, 1999). However, apart from onset, previous works manipulating set-size in visual search failed to provide empirical evidence for this kind of capture. By varying target-singelton distance method, based on a single set-size, we explored whether, in a serial search task, an attentional capture is triggered by static discontinuities such as those generated through the manipulation of color, form, and luminance. The results suggest that those physical properties are indeed able to capture attention automatically. PMID- 10814752 TI - Motion extrapolation is not responsible for the flash-lag effect. AB - To achieve perceptual alignment between a flashed target and a moving one, subjects typically require the flashed target to be aligned with a position that the moving target will only reach some time after the flash (the flash-lag effect). We examined how the magnitude of this misalignment changes near an abrupt change in velocity. The magnitude of the misalignment turns out to depend on the target's velocity after, rather than before, the flash. Thus, the misalignment cannot be caused by motion extrapolation. Neither can it be the inevitable consequence of a difference between the time it takes to process flashed and moving stimuli, because the magnitude of the misalignment is influenced by the extent to which subjects can anticipate the flash. We propose that it is the consequence of having to 'sample' the moving target's position in response to the flash. PMID- 10814753 TI - Comparing extra-retinal information about distance and direction. AB - The idea that extra-retinal information about the orientation of the eyes could be used to judge an object's distance has a long history, and has been the issue of considerable debate throughout this century. We here show that the poor performance in comparison with judgements of direction has geometrical rather than physiological reasons, and discuss why previous studies have misled us into believing that information about distance is even poorer than the geometry predicts. PMID- 10814754 TI - Opposite effects of GABA(A) and GABA(C) receptor antagonists on the b-wave of ERG recorded from the isolated rat retina. AB - The largest component in the fully dark-adapted ERG is a corneal-positive response, known as the b-wave, and believed to originate from depolarizing (ON type) bipolar cells. The two types of GABA receptors, GABA(A) and GABA(C) have been reported to exist on bipolar cells in rat retina. The goal of these experiments was to find whether these GABA receptors participate in the generation of the b-wave of electroretinogram (ERG). ERGs were recorded from the isolated rat retinas. The P(2)(t) component, obtained by subtracting the ERGs measured before the application of 50 micrograms APB from those measured after the application of 50 micrograms APB, was used as an indicator of depolarizing bipolar cell activity. Photovoltages, the fast P(3)(t) component of ERG, were registered between the two microelectrodes across the rod outer segments. Bicuculline and 3-aminopropylphosphonic acid (3-APA) were used as selective antagonists of GABA(A) and GABA(C) receptors, respectively. It was found that the GABA(A) and GABA(C) receptors antagonists have opposite effects on the b-wave: bicuculline increased the b-wave amplitude, while 3-APA reduced the amplitude of the b-wave. Neither bicuculline nor 3-APA affect photoreceptors. PMID- 10814755 TI - Motion-stereo mechanisms sensitive to inter-ocular phase. AB - We measured depth from interocular delay (The Pulfrich effect) using a dynamic random-dot pattern, consisting of a spatially-random noise field, the individual elements of which were sinusoidally-modulated in luminance over time. When an interocular phase difference in the flicker was introduced the display appeared to rotate in depth around a vertical axis like a transparent textured cylinder. The threshold phase lag was in the region of 5-10 degrees in different observers, which translated into a non-constant, decreasing interocular delay (ms) as the flicker frequency was increased. We conclude that phase, not delay, is the critical parameter in determining the detection of depth. Threshold signal/noise ratios were measured at different delays to determine the optimum phase difference, which was found to be in the region 60-90 degrees. However, delays centred around 180 degrees were less detectable than those around zero, ruling out a quadrature input to the stereo-motion mechanisms. We show that depth-from phase is a natural consequence of paired monocularly motion-direction sensitive neurones. Complex energy-detecting neurones are not required to explain the findings. PMID- 10814756 TI - Variance of high contrast textures is sensed using negative half-wave rectification. AB - A rectifying transformation is required to sense variations in texture contrast. Various theoretical and practical considerations have inclined researchers to suppose that this rectification is full-wave, rather than half-wave. In the studies reported here, observers are asked to judge which of two texture patches has higher texture variance. Textures are composed of small squares, with each square being painted with one of nine different luminances. Different texture variances are achieved by manipulating the histograms of the texture patches to be compared. When the nine luminances range linearly from 0 to 160 cd/m(2), the transformation mediating judgments of texture variance takes the form of a negative half-wave rectifier: texture variance judgments are determined exclusively by the frequencies of luminances below mean luminance in the textures being compared. However, when the nine luminances range linearly from 60 to 100 cd/m(2), two of three observers use a full-wave rectifying transformation in making texture variance judgments; the third observer continued to use a negative half-wave rectifier. The unexpectedly asymmetric roles played by low versus high luminances in texture variance judgments suggest that the off-center system may play a dominant role in human perception of texture contrast. PMID- 10814757 TI - Texture luminance judgments are approximately veridical. AB - This paper investigates intensity coding in human vision. Specifically, we address the following question: how do different luminances influence the perceived total luminance of a composite image? We investigate this question using a paradigm in which the observer attempts to judge, with feedback, which of two texture patches has higher total luminance. All patches are composed of nine luminances, ranging linearly from 0 (black) to a maximum luminance (white: 160 cd/m(2) in one condition; 20.2 cd/m(2) in another condition). Luminance histograms of the patches being compared are experimentally varied to derive, for each luminance nu, the impact exerted by texture elements (texels) of luminance nu on texture luminance judgments. We find that impact is approximately proportional to texel luminance; That is, a texture element exerts, on average, an impact on texture brightness (i.e. perceived texture luminance) that is proportional to its (the texel's) luminance. The only exception occurs for texels of maximal luminance, which surprisingly exert an impact that is slightly, but significantly, less than that exerted by texels of the next lower luminance. We conclude that visual intensity coding for purposes of assessing overall luminance of inhomogeneous patches is approximately veridical. In particular, texture luminance judgments are not mediated by a significant, compressive nonlinearity. PMID- 10814758 TI - The spectral sensitivities of the middle- and long-wavelength-sensitive cones derived from measurements in observers of known genotype. AB - The spectral sensitivities of middle- (M-) and long- (L-) wavelength-sensitive cones have been measured in dichromats of known genotype: M-cone sensitivities in nine protanopes, and L-cone sensitivities in 20 deuteranopes. We have used these dichromat cone spectral sensitivities, along with new luminous efficiency determinations, and existing spectral sensitivity and color matching data from normal trichromats, to derive estimates of the human M- and L-cone spectral sensitivities for 2 and 10 degrees dia. central targets, and an estimate of the photopic luminosity function [V(lambda)] for 2 degrees dia. targets, which we refer to as V(2)*(lambda). These new estimates are consistent with dichromatic and trichromatic spectral sensitivities and color matches. PMID- 10814759 TI - Tritanopic color matches and the middle- and long-wavelength-sensitive cone spectral sensitivities. AB - Tritanopic color matches (i.e. matches that depend on the middle- (M) and long- (L), but not short- (S) wavelength-sensitive cones) were made between two half fields: one illuminated by either a 405 or a 436 nm Hg spectral line; the other by a light of variable wavelength and radiance. Our purpose was to test between rival M- and L-cone spectral sensitivities, which should predict the tritanopic matches. The observers were tritanopes, in whom functioning S-cones are lacking, or normal trichromats, in whom artificial tritanopia was induced by a strong, violet adapting field. The wavelengths found to match the 405 and 436 nm lights agreed poorly with those predicted by the cone spectral sensitivities of Smith and Pokorny (1975) [Vision Research, 15, 161], while the 405 nm matching wavelength agreed poorly with that predicted by Stockman, MacLeod and Johnson (1993) [Journal of the Optical Society of America, A10, 2491]. Both matching wavelengths agreed well, however, with the predictions of the Stockman and Sharpe (2000) [Vision Research] M- and L-cone spectral sensitivities, which lie within the range of measured matches. PMID- 10814760 TI - Are there separate first-order and second-order mechanisms for orientation discrimination? AB - In a series of experiments we compared orientation discrimination performance for Gabor stimuli in which the stimulus profile was either matched to the receptive field profile of single V1 simple cells ('simple'), or in which the carrier and envelope orientations were different ('tigertails'). In the first Experiment, using small, high spatial frequency, peripheral stimuli to minimise the number of detectors involved, we found that simple stimuli were more detectable than tigertails of the same contrast energy, and that orientation discrimination thresholds for simple stimuli were lower than for tigertails of equal detectability. In later experiments with larger stimuli we measured thresholds for detecting tilts of the envelope with the carrier fixed in orientation. Envelope thresholds were similar for different carrier orientations, but carrier orientation had a strong biasing effect upon perceived envelope orientation. When the orientation difference between envelope and carrier was small, the carrier orientation was attracted to that of the envelope; when the difference was large (>10 degrees ) repulsion was found. The biases were reduced by half-wave rectifying the stimuli, putatively making the envelope visible to a first-order filter (Experiment 2). Discrimination thresholds for envelope orientation were higher than those for carrier orientation, and this difference was greater for briefly-presented parafoveal stimuli than for long duration foveal stimuli (Experiments 3 and 4). We conclude from these results that there are separate mechanisms for envelope and carrier orientation discriminations for large stimuli, but that first- and second-order mechanisms are not independent in the discrimination of orientation. PMID- 10814761 TI - Binocular disparity can explain the orientation of ocular dominance stripes in primate primary visual area (V1). AB - In the primate primary visual area (V1), the ocular dominance pattern consists of alternating monocular stripes. Stripe orientation follows systematic trends preserved across several species. I propose that these trends result from minimizing the length of intra-cortical wiring needed to recombine information from the two eyes in order to achieve the perception of depth. I argue that the stripe orientation at any point of V1 should follow the direction of binocular disparity in the corresponding point of the visual field. The optimal pattern of stripes determined from this argument agrees with the ocular dominance pattern of macaque and Cebus monkeys. This theory predicts that for any point in the visual field the limits of depth perception are greatest in the direction along the ocular dominance stripes at that point. PMID- 10814762 TI - A new test of contour integration deficits in patients with a history of disrupted binocular experience during visual development. AB - Previous studies have suggested that the integration of orientation information across space is impaired in amblyopia. We developed a method for quantifying orientation-domain processing using a test format that is suitable for clinical application. The test comprises a graded series of cards where each card includes a closed path (contour) of high contrast Gabor signals embedded in a random background of Gabor signals. Contour visibility in both normals and patients with histories of abnormal binocular vision depends jointly on the spacing of elements on the contour as well as background element density. Strabismic amblyopes show significant degradation of performance compared to normals. Small but significant losses in sensitivity were also observed in a group of non-amblyopic strabismus patients. Threshold measurements made with contrast reducing diffusers indicated that the amblyopic loss is not due to the reduced contrast sensitivity of the amblyopic eye. An abnormal pattern of long-range connectivity between spatial filters or a loss of such connectivity appears to be the primary source of contour integration deficits in amblyopia and strabismus. PMID- 10814763 TI - Electrical multisite stimulation of the isolated chicken retina. AB - Visual prostheses such as subretinal implants are intended for electrical multisite excitation of the retinal network. To investigate relevant issues like spatial resolution and operational range, we have developed an in vitro method using microelectrode arrays to stimulate isolated retinae. Ganglion cell activity in the chicken retina evoked by distally applied spatial voltage patterns consisted of fast bursts, transient inhibition and delayed discharges, and depended on the amount, location and spatial pattern of the injected charge. The response was altered or disappeared when synaptic transmission was blocked. Our results indicate that shape perception and object location can be partially achieved with subretinal electrical multisite stimulation. PMID- 10814765 TI - Understanding the heterogeneity of depression through the triad of symptoms, course and risk factors: a longitudinal, population-based study. AB - BACKGROUND: There is an ongoing research effort to test if depression is a homogeneous clinical syndrome and to identify valid and useful subtypes based on the number and nature of depressive symptoms. This study summarizes the patterns of depressive symptoms evident in a prospective study of the general population and examines the validity of potential subtypes by studying their course and etiologic heterogeneity. METHODS: A general population sample of 1920 adults (aged 18-96) from the Baltimore Epidemiologic Catchment Area (ECA) follow-up study (1981 to 1993/6) were examined. Data on diagnoses, symptoms, course and risk factors were collected using the Diagnostic Interview Schedule (DIS). Latent class analysis was applied to summarize symptom patterns. Course characteristics and risk factor profiles were compared among potential subtypes based on the number of symptom groups or symptom patterns. Logistic regression models were used to examine the etiologic heterogeneity among potential subtypes based on symptoms. RESULTS: The number of symptom groups gave the most efficient insight into differential etiologic processes. Severe depression (7-9 symptom groups) was associated with female gender, family history of depression but not with stressful life events before the onset of the first episode. Moderate (5-6 symptom groups) and mild depression (3-4 symptom groups) were associated with family history of depression, stressful life events before the onset, but not with female gender. The latent class model generated patterns of depressive psychopathology as follows: anhedonia, suicidal, psychomotor, and severely depressed subtypes. The Anhedonia subtype showed a course and risk factor profile distinct from the others. LIMITATIONS: The measurement of psychopathology was based on self-reported DIS interviews instead of psychiatric assessments. Recall or report bias cannot be excluded in the ascertainment of family history and stressful life events. CONCLUSIONS: Depression is heterogeneous, even below the threshold of syndromal diagnosis. The severity of an episode appears to be more informative than the pattern of symptoms, with the possible exception of a putative anhedonic subtype. PMID- 10814766 TI - Minor depressive disorder in the context of miscarriage. AB - BACKGROUND: Although minor depressive disorder is of considerable clinical and public health importance, it has received limited research attention relative to major depressive disorder. This study examines the incidence rate and relative risk for minor depressive disorder following miscarriage. METHODS: Using a cohort design we tested whether miscarrying women are at increased risk for an episode of minor depression (diagnosed based on research criteria proposed in Appendix B of DSM-IV) in the 6 months following loss. The miscarriage cohort consisted of women attending a medical center for spontaneous abortion (n=229); the comparison group was a population-based cohort of women drawn from the community (n=230). RESULTS: Among miscarrying women, 5.2% experienced an episode of minor depression, compared with 1.0% of community women. The overall relative risk for an episode of minor depression for miscarrying women was 5.2 (95% confidence interval, 1.2-23.6). Relative risk did not vary by length of gestation at the time of loss or attitude toward the pregnancy. The majority of episodes in miscarrying women began within 1 month following loss. LIMITATIONS: Minor depression was relatively rare in both study cohorts. The resulting limits on statistical power reduced our ability to identify factors, such as sociodemographic or reproductive history variables that might moderate the effect of miscarriage on risk for minor depression. CONCLUSIONS: These results, in the context of prior work showing increased risks of major depression and depressive symptoms following miscarriage, lend some support to the conceptualization of minor depressive disorder as part of a continuum of symptom severity. Miscarrying women should be evaluated for depression at their follow-up medical visits. PMID- 10814767 TI - Reconfirming the role of life events for the timing of depressive episodes. A two year prospective follow-up study. AB - BACKGROUND: Since the 1960s the association of stressful life events and depression seemed to be firmly established. However, a few recent studies did not confirm those earlier findings. One of the reasons discussed for the inconsistencies was the sampling of milder depressed neurotic out-patients in the earlier studies vs. more severely ill endogenous type in-patients in recent studies. METHODS: This investigation was carried out with 50 consecutively admitted in-patients with endogenous depression according to ICD 9 and unipolar major depression according to DSM-III-R as ascertained by SCID. The control sample consisted of 26 healthy volunteers. Life events and chronic distressing life conditions were recorded with the Munich Interview for the Assessment of Life Events and Conditions (MEL) every 3 months over a period of 2 years along with psychopathological symptoms and recurrencies. Hence the design was prospective in the sense that life events were recorded for one 3-month cross section, the depressive reaction for the subsequent one. BDI scores taken at the respective cross section were used to control for depressive bias of the subjective part of the patient's life event evaluation. RESULTS: Three months prior to the index hospitalization patients were more often affected by life events and conditions than controls. The number of stressful conditions prior to the index hospitalization indicated the time to relapse after discharge. Controls showed more desirable positive conditions than patients. Relapse patients suffered more often stressful life events and conditions than non-relapsers 3 months prior to their relapse. Multivariate analysis indicates that the cumulative number of life events within the 2-year course is the best predictor of the BDI score at the end of the follow-up period. LIMITATIONS: Since the subjective component of life event assessment by MEL displayed a higher impact on the course of depression than the objective part of the assessment, confounding of subjective ratings, attributional styles, and depressive symptoms may be a problem although controlled for in this study. CONCLUSION: The results support the importance of stressful life events and chronic distressing conditions for the 2-year course and outcome of major depression in an in-patient sample. Since the overall consistency of significant results was more pronounced in the subjective than in the objective part of the MEL the results fit best a circular pathogenetic model of interactions between life events, their individual evaluation by the patient, and depressive symptoms. PMID- 10814768 TI - Psychosocial correlates of prepartum and postpartum depressed mood. AB - BACKGROUND: The aim of the present study was to delineate the influence of maternal stress, social support and coping styles on depressed mood during pregnancy and the early postpartum period. METHODS: Beginning in the third month of pregnancy, data on numerous variables including daily stress (Hassles), state anxiety (STAI-state), pregnancy-specific stress (PEQ) and depressed mood (DACL) were collected monthly. In each trimester social support (SSQ), coping strategies (CISS) and pregnancy progress were assessed. Approximately 4-5 weeks following delivery, information on labor, delivery and infant status was collected and the DACL and the Edinburgh Postnatal Depression Scale (EPDS) were administered. The final sample consisted of 80 women. RESULTS: Approximately 16% of the women in this sample experienced depressed mood in the postpartum and 25% of the sample reported depressed mood only during pregnancy. Women depressed only during pregnancy and those depressed in the postpartum reported more emotional coping and higher trait and state anxiety during gestation. More hassles during pregnancy was related to prepartum depressed mood, but not postpartum depressed mood. Consistent with the literature, the best predictor of postpartum depressed mood was depressed mood during pregnancy. LIMITATIONS: The sample size was relatively small and we relied solely on self-reported depressive symptomology. CONCLUSIONS: The findings point to specific psychosocial variables which can be targeted early in pregnancy to reduce the rate of depressed mood in the prepartum and postpartum periods. PMID- 10814769 TI - Seasonality of birth in patients with mood disorders in Japan. AB - BACKGROUND: Findings about seasonality of birth in individuals with mood disorders have been inconsistent. METHODS: Data were collected from the governmental statistics, the Patient Survey in Japan in 1996. The number of patients with mood disorders was 13,969. We obtained information about each patient's date of birth, sex, and diagnosis according to ICD-10. Distributions of monthly birth numbers of patients with mood disorders were compared to those of the general population. RESULTS: Birth excess was observed from winter to early spring in both sexes, compared to births of the general population. The magnitude of the excess was larger in females than in males. Although the same tendencies were observed in patients with bipolar disorder and depressive disorder, the differences were more marked in females. LIMITATIONS: Insufficient birth data in the general population before 1940 and hospital diagnosis. CONCLUSION: Among Japanese patients with mood disorders, there are excess births from winter to early-spring, compared to the general population. This difference is more marked in females than in males. PMID- 10814770 TI - Seasonality, social zeitgebers and mood variability in entrainment of mood. Implications for seasonal affective disorder. AB - BACKGROUND: Seasonal variations in mood (seasonality) appear to be entrained to light, a physical zeitgeber. We hypothesised that people high in seasonality may be responsive to a range of zeitgebers, because of greater mood variability. We investigated whether the moods of people high in seasonality were more strongly entrained to the calendar week, a social zeitgeber, and whether any such effect was dependent on variability of mood. METHODS: 53 participants (14 male, 39 female; overall mean age=30) completed a daily mood report, over 56 consecutive days. Participants also completed the Seasonality Score Index (SSI) of the Seasonal Pattern Assessment Questionnaire. Each participant's time series of daily mood was analysed by spectral analysis to quantify the strength of their weekly mood cycle. RESULTS: Participants with high SSI scores (> or =11) had significantly stronger weekly mood cycles than those with low SSI scores (<11), and significantly greater variability in mood. Covarying for mood variability reduced the difference between high and low SSI groups in mean strength of weekly mood cycle to non-significance. LIMITATIONS: The time series of moods obtained was relatively short, and moods among high seasonal participants may have been affected by seasonal weather variability. CONCLUSIONS: People high in seasonality appear to be more responsive to external zeitgebers, and this could be linked to their greater variability in mood. The integration of research on mood variability with research on SAD appears to be warranted. PMID- 10814771 TI - Pretreatment plasma HVA and haloperidol response in acute mania. AB - INTRODUCTION: Pretreatment plasma homovanillic acid (HVA) levels have been reported to be a correlate of clinical response to typical antipsychotics for schizophrenic, bipolar manic, and mixed groups of psychotic patients. Biological markers of clinical response to antipsychotics could be useful for optimizing drug treatment. METHOD: Thirty-one consenting acute inpatient subjects between ages 19 and 66 years with a DSM-III-R clinical diagnosis of bipolar disorder, manic with psychotic features were entered into this double-blind study and were randomly assigned to receive either haloperidol 25 mg/day or haloperidol 5 mg for the 3-week study. Subjects also received one of the following concomitant medications: standard lithium, lorazepam 4 mg/day, or placebo. RESULTS: The primary multiple regression analysis, including all subjects on both haloperidol doses, yielded a significant main effect for pretreatment plasma HVA (n=31, F=5.7, P=0.025), indicating that higher pretreatment plasma HVA was predictive of better clinical response. In addition, the interaction between haloperidol dose and pretreatment plasma HVA was also significantly associated with clinical response (F=12.59, P=0.0015). When the two haloperidol doses were analyzed separately, we found that pretreatment plasma HVA was only correlated with clinical response in the low haloperidol 5 mg/day group (n=18, F=11.73, P=0.0038) and was unrelated to clinical response to the high haloperidol 25 mg/day group. LIMITATIONS: The sample size was small. Results may have been confounded by prior antipsychotic treatment and concomitant use of lithium or lorazepam. DISCUSSION: These results suggest that pretreatment plasma HVA could be useful for dosing antipsychotics. Patients with high plasma HVA levels would be good candidates for low-dose treatment because they are more likely to improve on such a dose, while patients with low plasma HVA levels might warrant more rapid dosage escalation. PMID- 10814772 TI - Depression in the carers of dementia sufferers: a comparison of the carers of patients suffering from dementia with Lewy bodies and the carers of patients with Alzheimer's disease. AB - BACKGROUND: Depression has not previously been studied in the carers of patients suffering from dementia with Lewy Bodies (DLB). METHOD: Twenty-five DLB carers were matched with 75 AD carers, according to carer characteristics. A standardised assessment incorporated measures of cognition, depression, psychosis and behavioural disturbance. DLB (consensus criteria), and AD (NINCDS ADRDA criteria) were diagnosed using operationalised clinical criteria. Carer depression was diagnosed using the Research Diagnostic Criteria. RESULTS: The overall frequency of depression was similar in DLB and AD carers (28% vs. 31%). DLB carers were significantly more likely to experience a major depressive disorder (3/25 - 12% vs. 0/75 - 0% chi(2)=9.3, P=0.002, df 1), although the numbers were small. The severity of behavioural disturbance was associated with carer depression in the DLB carers and in the total sample. CONCLUSION: The severity of behavioural disturbance was significantly associated with depression in DLB carers. PMID- 10814773 TI - Sertraline in the treatment of mixed anxiety and depression disorder. AB - BACKGROUND: Mixed anxiety and depression disorder (MAD) has been recognized in ICD-10 as a diagnostic group including those anxious and depressed patients which do not fit sufficient criteria for any major axis I disorders. MAD is usually treated as a combination of anxiety and depression, although there are data indicating that selective serotonin reuptake inhibitors (SSRIs) might be active on both anxiety and depression. METHOD: 38 patients diagnosed of MAD according to ICD-10 criteria were treated with flexible doses of sertraline for 8 weeks. Benzodiazepines were not allowed during the trial. Efficacy was evaluated with the Clinical Global Impression (CGI) improvement scale and with Hamilton's depression and anxiety Scales. Personality scales, including the Cloninger's TCI and Eysenck's EPQ, were used to test the predictive value of personality traits in the response to treatment. RESULTS: Anxiety was reduced by 55% and depression by 60% in Hamilton scales. At week 8, 29 patients were considered responders (CGI 1 o 2). Two patients discontinued the trial, only one of them due to adverse events. The mean dose of sertraline was 83.4 mg/day. CONCLUSION: Sertraline showed an excellent tolerability in patients with mixed anxiety-depression disorder despite high levels of baseline anxiety. The response level was high and similar to that reported for patients with major depression. These results warrant further controlled trials to assess the efficacy of SSRIs in MAD. PMID- 10814774 TI - Pituitary-adrenal function in adolescent psychiatric patients: impact of depressive symptoms. AB - BACKGROUND: The aim of this study was to determine whether depressive symptoms affect pituitary-adrenal function in adolescents, as they do in adults. METHODS: Salivary cortisol was measured before and after physical exercise in 23 hospitalized adolescent psychiatric patients and 13 age- and sex-matched healthy controls in a placebo-controlled design. In patients, cortisol profiles were assessed from 08:00 to 20:00 h before and after administration of low doses of dexamethasone or the natural steroid hydrocortisone. Patients were classified according to DSM III-R criteria and assigned to either a depressed group (n=10) or a non-depressed group (n=13). Subjective depressive symptoms were rated with the 90-item symptom checklist. RESULTS: Physical exercise increased cortisol output significantly in all subjects, but there were no differences between groups. In patients, no differences in basal cortisol levels were found between the depressed and non-depressed groups. Dexamethasone, but not hydrocortisone, was able to suppress cortisol levels in both groups. Differences between groups were only found in total cortisol output over the 3 days when data were analyzed according to the patients 'subjective' feelings of depression, with the highest cortisol levels in the 'subjectively more depressed' patients. CONCLUSIONS: The results obtained only partially support the hypothesis that depressive symptoms in adolescent psychiatric patients determine pituitary-adrenal (mal)function, and appear to depend on the strategy used. PMID- 10814775 TI - Preliminary evidence of an association between increased REM density and poor antidepressant response to partial sleep deprivation. AB - BACKGROUND: One night of total sleep deprivation or of late-night partial sleep deprivation (PSD) produces a temporary remission in approximately 40-60% of patients with major depressive disorder; however, little is known about polysomnography (PSG) characteristics of responders to these types of sleep deprivation (SD). METHODS: Twenty-three unmedicated unipolar patients (17-item Hamilton Depression Rating Scale (HDRS17) >16) and 14 normal controls underwent 1 night of late-night PSD (awake after 3 a.m.) Subjects underwent baseline PSG and received the HDRS17 at standard times before and after PSD. Clinical response was defined as a reduction of >30% in the modified HDRS17 (omitting sleep and weight loss items) following PSD. RESULTS: The 12 responders and 11 nonresponders did not differ from each other significantly on baseline HDRS17 or PSG variables. The only PSG variable correlating with percent decrease in modified HDRS17 was baseline REM density (Pearson's r=-0.52, n=23, P=0.01.) In other words, the lower the baseline REM density, the more robust the antidepressant response was. LIMITATIONS: Subject numbers are relatively small. CONCLUSIONS: Increased REM density, which reflects the number of rapid eye movements per epoch of REM sleep, may be a physiological marker for severity or poor prognosis in a variety of psychiatric disorders, including relapse in recovering alcoholics, suicidality in schizophrenia, and poor response to PSD or interpersonal psychotherapy in depression. PMID- 10814776 TI - Expression of human IAP-like protein in skeletal muscle: a possible explanation for the rare incidence of muscle fiber apoptosis in T-cell mediated inflammatory myopathies. AB - In Polymyositis (PM) and sporadic Inclusion Body Myositis (s-IBM), the CD8(+) cytotoxic T cells invade the muscle membrane and release perforin and granzyme B to induce cell death. Although granzyme B is a direct activator of executioner caspases, there is no convincing evidence of apoptosis in the muscle fibers of these patients. To search for an explanation, we examined the muscle expression of the human IAP-Like Protein (hILP), an evolutionarily conserved cell death suppressor, that exerts major anti-apoptotic effects by inhibiting the executioner caspases. Muscle biopsy specimens from patients with inflammatory myopathies and controls were studied with: (a) immunocytochemistry using antibodies against hILP and caspase-3 in single and double-labeled confocal laser microscopy; (b) immunoblotting of muscle extracts immunoreacted with anti-hILP antibodies; and (c) subcellular fractionation of muscle lysates immunoreacted with antibodies against hILP. We found that hILP is expressed on the sarcolemmal region and co-localizes with dystrophin. Caspase-3 is undetectable. Subcellular fractionation of the muscle specimens confirmed that hILP is a membrane associated protein. By immunoblotting, the 57 kD hILP was abundantly expressed in the normal as well as the diseased muscles. We conclude that in s-IBM and PM the expression of hILP, a major cell death suppressor, on the muscle membrane may prevent the induction of apoptosis by the autoinvasive cytotoxic T cells on the cell surface, by inhibiting the caspase activation. PMID- 10814777 TI - Antisense inhibition of pro-opiomelanocortin and proenkephalin A messenger RNA translation alters rat immune cell function in vitro. AB - Pro-opiomelanocortin (POMC) and proenkephalin A (PEA) antisense oligodeoxynucleotides respectively reduced and enhanced proliferation of rat splenocytes incubated with concanavalin A in vitro. Nonsense base sequences used as controls were without effect. Coincubation with the exogenous synthetic opioid peptides, ACTH, beta-endorphin, met-enkephalin or [D-ala,D-leu]-enkephalin did not significantly alter either the POMC or PEA antisense response, indicating potential differences in bioactivity of immunocyte opioid peptides compared with synthetic equivalents. Levels of the POMC opioid products, ACTH and beta endorphin, were significantly reduced in splenocytes incubated with POMC antisense probes. These data provide evidence for functional effects of endogenous opioid peptides on rat splenocyte proliferation in vitro. PMID- 10814778 TI - Differential expression of stress proteins in human adult astrocytes in response to cytokines. AB - Various lines of evidence suggest a close relationship between heat shock proteins (hsp) and several autoimmune diseases such as arthritis, diabetes and multiple sclerosis. While enhanced expression of hsp in autoimmune diseases is often regarded as a non-specific bystander effect of the inflammatory process, surprisingly little is known on hsp regulation by inflammatory mediators such as cytokines. In this study cytokine-induced expression of hsp60, hsp27 and alphaB crystallin was studied in cultures of primary human adult astrocytes at the mRNA as well as at the protein level. We show differential hsp expression patterns in response to pro-inflammatory and immunoregulatory cytokines. Hsp60 expression was found to be enhanced in response to cytokines as diverse as IL-1beta, TNF-alpha, IL-4, IL-6 and IL-10. Upregulation of hsp27, however, was primarily induced by immunoregulatory cytokines like IL-4, IL-6 and TGF-beta whereas alphaB-crystallin expression was found to be enhanced by the pro-inflammatory cytokine TNF-alpha only. None of the cytokines studied was able to enhance expression of all three hsp simultaneously. These results show that in human astrocytes induced expression of hsp27 and alphaB-crystallin is dependent on the presence of a defined set of stimuli, while induced expression of hsp60 is a much less selective event. This highly differential pattern of hsp expression in response to inflammatory mediators known to play an important role in the pathogenesis of autoimmune diseases indicates that hsp responses are specific rather than non specific bystander responses. PMID- 10814779 TI - Modulation of Alzheimer's beta-amyloid neurotoxicity by site-directed single chain antibody. AB - A single-chain antibody was constructed from variable regions of heavy and light genes of the parental anti-beta-amyloid peptide IgM 508 antibody. This antibody exhibits anti-aggregating properties, leading to disaggregation of Alzheimer beta amyloid (betaA) fibrils and prevents its toxic effect on cultured PC-12 cells. Sequencing of the small antibody, namely 508 (Fv), revealed that the V(L) domain contained a cysteine residue in the complementary determining region (CDR)3 (residue 96) which affects its solubility and stability. The cysteine codon was replaced using SOE PCR, and one of the mutants obtained, namely 508F(Fv) (containing phenylalanine instead of cysteine), showed an increased storage stability and higher affinity compared to the wild type. Antibody 508F(Fv) prevents the neurotoxicity of betaA (90% cell viability) and disrupts the fibril structure of beta-amyloid (62% decrease in ThT fluorescence). The ability of antibody 508F(Fv) to dissolve already-formed betaA fibrils makes it a good candidate for intracellular expression and modulation of APP processing as the first step towards the production of therapeutic protection molecules for Alzheimer's disease treatment. PMID- 10814780 TI - Interleukin 1-beta modulates the effects of hypoxia in neuronal culture. AB - In order to study the role of interleukin-1beta (IL-1beta) in homeostasis, hypoxia and recovery of neuronal cells, we studied the expression and release of tumor necrosis factor-alpha (TNF-alpha) and nerve growth factor (NGF), in relation to the presence or absence of this cytokine in culture medium. Moreover, we evaluated cell mortality in the same conditions. For this aim, we used untreated and IL-1beta pre-immunoneutralized hippocampal neuronal cultures exposed to mild hypoxic stress and left to reoxygenate. Semiquantitative reverse transciptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) determined gene expression and protein levels. Mild hypoxic stress provokes a decrease in both the expression and release of TNF-alpha and NGF. IL 1beta neutralization results in an inversion of this pattern since treated hypoxic cultures exhibited an increase of both expression and release of NGF. In pretreated hypoxic cells the increased expression of TNF-alpha was not followed by a rise in release. Reoxygenation reversed the observed effects in both cultures and the levels of cytokine expression and release were approaching control values. Our data show that in physiological conditions IL-1beta may have a neuroprotective action through positive modulation of NGF. Contrary to that, in presence of insult, IL-1beta may have an opposite role, since neutralization provoked an increase of expression and release of NGF. In addition, we demonstrated that neuronal cells are biochemically capable, not only of maintaining and recovering the homeostasis, but also of activating the appropriate response to insult. IL-1beta may have a pivotal role in this mechanism through the modulation of NGF and to a lesser degree of TNF-alpha. PMID- 10814781 TI - CD8(+) T cells from Theiler's virus-resistant BALB/cByJ mice downregulate pathogenic virus-specific CD4(+) T cells. AB - Theiler's murine encephalomyelitis virus (TMEV) is a picornavirus which induces an immune-mediated demyelinating disease in susceptible strains of mice and serves as a relevant animal model for multiple sclerosis. Treatment with low dose irradiation prior to infection with the BeAn strain of TMEV renders the genetically resistant BALB/cByJ (C/cByJ) mice susceptible to disease. Previous studies have shown that disease resistance in the C/cByJ is mediated by a 'regulatory' CD8(+) T cell population, which does not appear to function via a cytolytic mechanism. We show here that TMEV-specific CD4(+) T cell blasts transferred into susceptible, irradiated C/cByJ accelerate clinical disease and enhance TMEV-specific DTH and proliferation in these animals. Significantly, CD8(+) cells from infected, resistant C/cByJ mice specifically downregulate the in vivo disease potentiation and diminish virus specific DTH, and proliferative and pro-inflammatory cytokine responses (IFNgamma and IL-2) in recipients of TMEV specific CD4(+) T cell blasts. These results indicate that TMEV infection of resistant C/cByJ mice induces a radiosensitive population of regulatory CD8(+) T cells which actively downregulate inherent Th1 responses which have disease initiating potential. PMID- 10814782 TI - Novel opioid peptides endomorphin-1 and endomorphin-2 are present in mammalian immune tissues. AB - Endomorphin (EM)-1 and EM-2 are opioid tetrapeptides, reported within the central nervous system, which have very high specificity and affinity for the mu-opioid receptor. We have used newly developed and well-characterised radioimmunoassays (RIAs) in combination with reversed-phase high-performance liquid chromatography (HPLC) to detect EM-1 and EM-2 immunoreactivity (ir) in rat immune tissues. Endomorphins were detectable in extracts of rat spleen (total EM-1-ir/spleen: 440+/-73 pg, mean+/-SEM, a=group of eight rats; EM-2-ir: 150+/-12 pg) and thymus (EM-1-ir: 152+/-18 pg, mean+/-SEM n=8; EM-2-ir: 156+/-28 pg). EM-2-ir was detectable in extracts of human spleen (338+/-196 pg/g tissue, n=3). Multiple peaks of EM-1-ir and EM-2-ir were observed in rat spleen and thymus extracts, and multiple peaks of EM-2-ir were observed in extracts of human spleen, following reversed-phase HPLC and RIAs. This is the first report of endomorphin immunoreactivity in tissues of the rat and human immune systems. PMID- 10814783 TI - A role for alpha4-integrin in the pathology following Semliki Forest virus infection. AB - Migration of cells into the central nervous system (CNS) is a pivotal step in the pathogenesis of immune-mediated diseases such as multiple sclerosis (MS), experimental allergic encephalomyelitis (EAE) and virus-induced demyelinating diseases. Such migration is dependent on expression of adhesion molecules. The expression of adhesion molecules in the CNS was studied in Biozzi ABH mice infected with Semliki Forest virus (SFV) A7(74) - an important demyelinating model of MS. Expression of LFA-1alpha/CD11a, LFA-1beta/CD18 and ICAM-1/CD56 were rapidly elevated and remained high whereas MAC-1, CD44 and VCAM-1/CD106 were less widely expressed. The alpha4-integrin VLA-4/CD49d was more specifically associated with CNS lesions. To identify the importance of VLA-4, CD44, ICAM-1 and MAC-1 in the pathogenesis of SFV infection, monoclonal antibodies that block these adhesion molecules were administered in vivo during infection. Anti-VLA-4 treatment dramatically reduced the cellular infiltrates and demyelination within the CNS but did not affect the clearance of virus while antibodies to CD44, ICAM and MAC-1 antibody treatment had no effect. This study demonstrates that SFV infection induces the expression of adhesion molecules within the CNS and that VLA-4 plays an important role in the development of inflammation and demyelination in the CNS following SFV infection. PMID- 10814784 TI - Role of astrocytes in antigen presentation and naive T-cell activation. AB - Astrocytes may have a role in antigen presentation in inflammatory diseases of the central nervous system (CNS) such as MS and EAE. In this study, we have assessed whether purified astrocyte cultures could stimulate naive CD4(+) or CD8(+) T-cells from TCR transgenic mice. As previously described, astrocytes sustained antigen-specific CD4(+) T-cell proliferation only in the presence of IFN-gamma, which promotes expression of both MHC class II and B7 molecules on astrocytes. In addition, we show that astrocytes also have the capacity to present antigens to naive CD8(+) T-cell and promote their proliferation. In one system, this CD8(+) T-cell proliferation was dependent on IFN-gamma-induced upregulation of B7 molecules on astrocytes. However, in a second TCR transgenic system, astrocytes could induce naive CD8(+) T-cell proliferation even in the absence of IFN-gamma. The possible implications of these findings for the pathophysiology of CNS inflammatory diseases are discussed. PMID- 10814785 TI - Haemopoietic biglycan produced by brain cells stimulates growth of microglial cells. AB - We have recently found that soluble biglycan purified from rat thymic myoid cells had haemopoietic activity capable of inducing preferential growth and differentiation of monocytic lineage cells from various haemopoietic sources, including brain microglial cells. In the present study, to understand developmental mechanisms of microglial/monocytic cells in the brain, we have attempted to identify haemopoietic activity of the brain biglycan. The mRNA and the immunological epitope of biglycan were detected in the rat brain homogenates and several rat glial cell lines. Immunohistochemical study showed that several different types of brain cells produced biglycan. During development biglycan synthesis in the brain appeared to be increased. The brain haemopoietic biglycan was easily separated by DEAE-Sepharose chromatography from the macrophage colony stimulating factor (M-CSF) which was concomitantly produced from the brain cells. The brain haemopoietic biglycan, purified through immunoaffinity column, indeed stimulated growth of primarily cultured microglial cells. Taken together, these results suggest that the haemopoietic biglycan plays an important role in generating brain-specific circumstances for development of microglial/monocytic cells. PMID- 10814787 TI - Interleukin-4-inhibited mRNA expression in mixed rat glial and in isolated microglial cultures. AB - Interleukin-4 (IL-4) likely is one of the key players in the concert of immunosuppressive factors in brain. Therefore, influences of the cytokine on mRNA expression of endogenous mediators of inflammation, such as cyclooxygenase-2 (COX 2), inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF alpha), interferon-inducible protein 10 (IP-10), interleukin-3 receptor-beta (IL 3R-beta), and of another antiinflammatory cytokine, IL-10, have been evaluated in the present study by semi-quantitative RT-PCR. Primary rat mixed glial cultures and isolated microglial cells, the resident immunocytes of the brain, have been used as rich sources of these mRNAs in response to the bacterial cell wall component lipopolysaccharide (LPS). Time-course studies showed peak levels of LPS increased mRNAs at approximately 4 h. Interestingly, IL-10 mRNA was elevated also upon the LPS-stimulus. IL-4, given 30 min before LPS, inhibited increases of all mRNAs significantly, including IL-10 mRNA. IL-4, however, induced peroxisome proliferator-activated receptor (PPAR)-gamma in cultured microglia. This induction was completely inhibited by simultaneous administration of LPS. The data confirms IL-4 as an important antiinflammatory cytokine and gives some idea of cross-talk between intracellular signaling evoked by pro- and antiinflammatory substances. PMID- 10814786 TI - Inducible expression of the long pentraxin PTX3 in the central nervous system. AB - PTX3 is a prototypic long pentraxin consisting of a C terminal 203-amino acid pentraxin-like domain coupled with an N-terminal 178-amino acid unrelated portion. PTX3 is induced by primary proinflammatory signals in various cell types, most prominently macrophages and endothelial cells. Other long pentraxins, such as murine or rat neuronal pentraxin 1 (NP1) and human neuronal pentraxin 2 (NPTX2), are expressed in the central nervous system (CNS). The present study was designed to investigate whether PTX3 is expressed in the brain and to define the structures and cells involved. Intracerebroventricular (i.c.v.), but not i.v., injection of LPS induced high levels of PTX3 mRNA in the mouse brain. In contrast NP1 is constitutively expressed in the murine CNS and is not modulated by LPS administration. I.c.v. IL-1beta was also a potent inducer of PTX3 expression in the CNS, whereas TNFalpha was substantially less effective and IL-6 induced a barely detectable signal. Central administration of LPS and IL-1 induced PTX3 also in the periphery (heart), whereas the reverse did not occur. Expression of PTX3 was also observed in the brain of mice infected with Candida albicans (C. albicans) or Cryptococcus neoformans. (C. neoformans). The kinetics of PTX3 gene induction were consistently different between C. albicans- and C. neoformans infected mice, according to the diverse outcome of the CNS immune reaction. In situ hybridization revealed that i.c.v. injection of LPS induced a strong PTX3 expression in presumptive glial cells, in the white matter (corpus callosum, fimbria) and meningeal pia mater as well as in dentate gyrus hilus and granule cells. No constitutive expression of PTX3 was detected. Central expression of PTX3 may amplify mechanisms of innate resistance and damage in the CNS. The possibility of a direct interaction of PTX3 with neuronal cells, as suggested for NPTX2, remains to be explored. PMID- 10814788 TI - A rat model for human T lymphocyte virus type I-associated myeloneuropathy. down regulation of bcl-2 expression and increase in sensitivity to TNF-alpha of the spinal oligodendrocytes. AB - We reported that the tumor necrosis factor-alpha (TNF-alpha) expression and apoptotic death of oligodendrocytes appeared to be a major pathogenesis of the demyelination of spinal cords of Wistar-King-Aptekman-Hokudai (WKAH) rats with human T lymphocyte virus type I (HTLV-I) infection, HAM rats. In the present study, we examined the sensitivity to TNF-alpha-induced cell death of in vitro separated oligodendrocytes from HTLV-I-infected WKAH rats. Although the number of non-viable oligodendrocytes increased by adding recombinant TNF-alpha, in a dose dependent manner, in both HTLV-I-infected and uninfected control rats, oligodendrocytes from the infected rats were more susceptible to TNF-alpha. In situ detection of DNA fragmentation showed apoptotic death of oligodendrocytes. The expression of bcl-2, an anti-apoptotic gene, was strongly down-regulated in oligodendrocytes of the infected rats but not in the control rats. We suggest that the down-regulation of bcl-2 expression in the oligodendrocytes of the HTLV I-infected rats may increase the susceptibility to TNF-alpha-induced apoptosis of oligodendrocytes, the result being development of HTLV-I-induced myeloneuropathy in rats. PMID- 10814789 TI - Spatiotemporal induction patterns of cytokine and related immune signal molecule mRNAs in response to intrastriatal injection of lipopolysaccharide. AB - The brain's response to a direct immune challenge was examined by in situ hybridization histochemistry. Lipopolysaccharide (bacterial endotoxin) injected acutely into rat striatum induced mRNA expression for inhibitory factor kappaBalpha, interleukin (IL)-1beta, tumor necrosis factor-alpha, IL-6, IL-12 p35, inducible nitric oxide synthase, IL-1 receptor antagonist, and the type 1 IL 1 receptor. Expression patterns were evaluated at select time points ranging from 15 min to 3 days post-injection. Rats injected with vehicle alone were used to control for mechanical effects. Following lipopolysaccharide administration, a wave of mRNA induction within brain parenchyma radiated outward from the injection site, generally peaking in intensity at the 16-h time point. The individual profiles of cytokine mRNA induction patterns reveal that the brain's immune response to local inflammatory stimulation is quite elaborate and in many ways resembles the progression of cytokine induction customary of localized inflammation in peripheral tissues. PMID- 10814790 TI - Tumour necrosis factor-alpha has few morphological effects within the dorsal columns of the spinal cord, in contrast to its effects in the peripheral nervous system. AB - There is circumstantial evidence implicating the pro-inflammatory cytokine tumour necrosis factor (TNF) in the pathogenesis of multiple sclerosis (MS), but there is no direct evidence that TNF can produce demyelination in the central nervous system (CNS). We demonstrate here that single injections of TNF into the dorsal columns of adult rats produced a mild inflammatory response indistinguishable from that seen in control cords, but did not induce demyelination. A similar response was seen when TNF-alpha was injected into dorsal columns where central axons had been remyelinated by Schwann cells. In marked contrast, single intraneural injections of TNF into sciatic nerves produced acute changes in the endoneurial microvascular bed that were followed by demyelination and degeneration. PMID- 10814791 TI - Normal polyclonal immunoglobulins ('IVIg') inhibit microglial phagocytosis in vitro. AB - Phagocytosis removes pathogens and tissue debris during inflammatory reactions, but also plays an important role in autoimmune reactions. The main phagocytes in the central nervous system (CNS) are microglial cells that are activated during CNS inflammation. In the treatment of inflammatory demyelinating diseases like multiple sclerosis (MS), administration of intravenous immunoglobulins (IVIg) has become a promising immunomodulatory therapy. Although a large number of potential mechanisms for the effects of IVIg has been suggested, the precise mode of action in CNS inflammation is unknown. We assessed the influence of IVIg on phagocytosis and endocytosis in microglia in vitro. IVIg had little effect on non-specific phagocytosis of latex particles in untreated microglia, while there was a dose dependent inhibition in microglia activated with LPS and IFNgamma. Endocytosis of soluble myelin basic protein (MBP) was downregulated by IVIg in both untreated and activated microglia. The effect was mediated by an F(ab')(2) preparation of immunoglobulins, suggesting that Fc receptor-mediated phagocytosis is not involved. Intact IVIg, but not F(ab')(2) fragments also suppressed Fc receptor mediated phagocytosis of opsonised erythrocytes in both untreated and activated microglia. These results show that IVIg can inhibit the phagocytic activity of microglia via different mechanisms. Such an effect could contribute to the immunomodulatory capacity of IVIg in inflammatory CNS diseases. PMID- 10814792 TI - Targeting antigen-specific T cells by genetically engineered antigen presenting cells. A strategy for specific immunotherapy of autoimmune disease. AB - We describe a strategy for specific immunotherapy of autoimmune disease based on targeting the antigen-specific T cells in an experimental model of myasthenia gravis. To address the problem of heterogeneity of the T cell repertoire, we have genetically engineered antigen presenting cells (APCs) to process and present epitopes of the autoantigen, acetylcholine receptor (AChR), to the entire spectrum of AChR-specific syngeneic T cells. APCs derived from BALB/c mice were stably transfected with cDNA for the key immunogenic domain of the AChR alpha subunit, flanked by sequences of the lysosome-associated membrane protein (LAMP) that direct APCs to process and present the antigen via the MHC Class II pathway. Transfected APCs strongly stimulated AChR-specific T cells from BALB/c mice. Fas ligand, or antibody to Fas, abrogated the T cell response, by inducing apoptosis of the APC-stimulated T cells. The new results of this investigation are (1) that autoreactive T cells can be effectively targeted by autologous APCs that are engineered to present the relevant autoantigen, and (2) that these specifically targeted and activated T cells can be profoundly inhibited by agents that trigger the Fas-mediated apoptosis pathway. The present findings suggest that engineering APCs for simultaneous presentation of the autoantigen and delivery of FasL will provide a powerful strategy for the elimination of autoreactive T cells. PMID- 10814793 TI - Selection of anti-myelin basic protein T-cell lines in the lewis rat: V-beta 8.2 dominance and conserved complementarity-determining-region-3 motifs are dependent on serine at position 78 of myelin basic protein. AB - In the Lewis rat, the dominant T cell repertoire to myelin basic protein (MBP) is directed to the peptide 71-87 and the T cell receptors of pathogenic T cells are of the Vbeta 8.2 genotype with short CDR3 sequences having a characteristic motif. However, this paradigm has been reached through analysis of long-term encephalitogenic lines and clones. We initiated the present study to examine the process of selection of the TCR Vbeta 8.2 and characteristic CDR3 motifs upon immunization with guinea-pig MBP, and rat or guinea-pig 71-87 peptides. We found that the dominance of Vbeta 8.2 developed progressively over 4-6 in vitro stimulations. Following immunization with rat 70-86, which differs from the guinea-pig peptide in one amino acid at position 78, the dominance of Vbeta 8.2 and the characteristic CDR sequences are not seen. Thus, Vbeta 8.2 dominance and specific CDR3 TCR motifs are seen with heterologous GpMBP but not with self rat MBP. PMID- 10814794 TI - Coexpression of Fas/FasL and Bax on brain and infiltrating T cells in the central nervous system is closely associated with apoptotic cell death during autoimmune encephalomyelitis. AB - Recent studies have suggested that autoimmune inflammation elicited in the central nervous system (CNS) is subsided by apoptotic cell death of inflammatory cells. To elucidate the molecular mechanism of apoptosis of infiltrating T and other cells occurring in the CNS during autoimmune encephalomyelitis, we determined the type of apoptotic cells and the localization of apoptosis-related molecules (Fas, FasL, Bax, Bcl-2 and active caspase 3) by immunohistochemistry. Double labeling with the TUNEL method and cell-type markers showed that infiltrating T cells and microglia/macrophages underwent apoptosis, while astrocytes and neurons did not. Staining for apoptosis-related molecules revealed that infiltrating T cells and microglia/macrophages, but not astrocytes and neurons, expressed both Fas-FasL and Bax. The distribution and cell type of active caspase 3-positive cells were essentially the same as those of TUNEL positive cells. These findings suggest that coexpression of Fas/FasL and Bax is closely associated with apoptotic cell death of infiltrating T cells and microglia in the CNS. Furthermore, astrocytes which express Fas and FasL, but not Bax, may play an important role in regulating inflammation in the CNS by inducing apoptotic cell death of infiltrating T cells and microglia, both of which have an inflammation-promoting nature. PMID- 10814795 TI - Saturable entry of leukemia inhibitory factor from blood to the central nervous system. AB - Leukemia inhibitory factor (LIF) is a neurotrophic cytokine now under clinical investigation for its effects on the CNS. We studied its passage across the blood brain barrier (BBB) from blood to brain and spinal cord. Although a large amount of LIF was reversibly associated with the cerebral vasculature, intact LIF did reach brain parenchyma. Multiple-time regression analysis showed ready access of LIF to the CNS at a rate much faster than that of the vascular marker albumin. Excess LIF inhibited the entry of 125I-LIF after administration i.v. or by in situ perfusion in blood-free buffer. Efflux of LIF from brain to blood was slower than reabsorption by CSF bulk flow, indicating that LIF tended to be retained in the brain. Although ciliary neurotrophic factor (CNTF) and LIF bind to the same receptor complex, CNTF did not cross-inhibit the entry of LIF into the CNS. A monoclonal antibody to LIF, however, abolished the entry of LIF. Our results show that peripherally administered LIF readily enters the brain and spinal cord by a saturable transport system across the BBB that may have biological implications. PMID- 10814796 TI - The cellular response of JC virus T-antigen-induced brain tumor implants to a Murine intra-ocular model. AB - In order to define the immunologic response to central nervous system tumors in a controlled fashion, we compared xenogeneic, allogeneic and syngeneic transplants of JC virus-induced neural tumor cell aggregates implanted into anterior ocular chambers of mice. Semiquantitative assessment of the level of leukocyte common antigen (CD45) of the transplants by immunohistochemistry was used to gauge rejection. Reticulin staining was used to monitor vascularization. Immunoreactivity to the viral oncoprotein, T-antigen, was confirmed by immunohistochemistry and immunoprecipitation/Western blot analysis. The results demonstrated that transplants were viable at all time-points and developed vascularization as early as three days after transplantation. Xenotransplants, 13 days post-transplantation, and allogeneic transplants, 25 days post transplantation were infiltrated with polymorphonuclear leukocytes. Fewer CD45 positive cells were demonstrated in syngeneic transplants. High levels of JCV T antigen stimulated rejection in syngeneic transplants. These results establish a model for further investigation of the natural and induced immunologic response to central nervous system tumors. PMID- 10814797 TI - Autoimmune T cells retard the loss of function in injured rat optic nerves. AB - We recently demonstrated that autoimmune T cells protect neurons from secondary degeneration after central nervous system (CNS) axotomy in rats. Here we show, using both morphological and electrophysiological analyses, that the neuroprotection is long-lasting and is manifested functionally. After partial crush injury of the rat optic nerve, systemic injection of autoimmune T cells specific to myelin basic protein significantly diminished the loss of retinal ganglion cells and conducting axons, and significantly retarded the loss of the visual response evoked by light stimulation. These results support our challenge to the traditional concept of autoimmunity as always harmful, and suggest that in certain situations T cell autoimmunity may actually be beneficial. It might be possible to employ T cell intervention to slow down functional loss in the injured CNS. PMID- 10814798 TI - Focal accumulation of cyclooxygenase-1 (COX-1) and COX-2 expressing cells in cerebral malaria. AB - Intravascular sequestration and altered cytokine expression patterns are key determinators of CNS lesion formation in patients with cerebral malaria (CM). Among others, altered prostaglandin concentrations were revealed by clinical trials in peripheral blood of CM patients. Prostaglandin synthesis is controlled by cyclooxygenases (COX, prostaglandin endoperoxide synthase, PGG/H synthase) and COX expression has been attributed a key role in immunomodulation, hemostasis and inflammation in a wide variety of pathologically altered brain tissues. We have now analyzed expression of COX-1 and COX-2 in brains of patients with CM by immunohistochemistry. Double labeling experiments were used to verify the cellular identity of COX-1 and COX-2 expressing cells. Compared to healthy controls, significant (P=0.0006) accumulation of COX-1 expressing macrophages/microglial cells was detected in Durck's granulomas. Accumulations of COX-2 expressing endothelial cells (P=0.0006) and COX-2 expressing astrocytes (P=0.0012) were detected in CM brain parenchyma. The restricted expression and accumulation of COX-1 and COX-2 in CM brains adds convincing evidence for the participation of cyclooxygenases in the formation of fever, inflammation and granuloma in these patients. PMID- 10814799 TI - Critical illness polyneuropathy and myopathy (CIPNM): evidence for local immune activation by cytokine-expression in the muscle tissue. AB - In a longitudinal prospective study a muscle biopsy was taken from 30/32 (33%) of the 98 patients who developed critical illness polyneuropathy and myopathy (CIPNM). Neuropathic changes were found in 37%, myopathic in 40%, and a combination in 23% of the biopsies. The immunohistopathology showed macrophages and Th-cells in 40% and 60% of the muscle biopsies respectively. Small mainly perivascular infiltrates contained macrophages and Th-cells. ICAM-1, VCAM and MAC were found on the vascular endothelium in 58%, 53% and 79% respectively. In all biopsies there was an upregulation of both HLA-I and HLA-DR. Proinflammatory cytokines and TNFalphaR75 were also produced locally (IL-1beta in 71%, IFN-gamma in 40%, IL-12 in 73%, TNFalphaR75 in 90%). The anti-inflammatory cytokine IL-10 was simultaneously expressed in 96% of the biopsies. HLA-DR, TNFalphaR75 and IL 10 differed significantly when compared with control muscle biopsies. Our data provide evidence that small numbers of activated leukocytes producing both pro- and anti-inflammatory cytokines infiltrate skeletal muscle of CIPNM patients. We propose that the local balance of leukocyte activities is of importance in the pathophysiology of muscle weakness in CIPNM. PMID- 10814800 TI - Localization of the human SP3 gene to chromosome 7p14-p15.2. The lack of expression in multiple sclerosis does not reflect abnormal gene organization. AB - Sp3 belongs to a large family of transcription factors that bind GC/T box elements. We have previously demonstrated the deficient expression of Sp3 in peripheral blood mononuclear cells (PBMC) from most patients with multiple sclerosis (MS). In the current study, the Sp3 gene was assigned to chromosome 7 by using somatic cell hybrid mapping and analysis of a chromosome 7-specific cosmid library. The position of Sp3 was refined to 7p14-p15.2 by fluorescence in situ hybridization (FISH). Southern blot and polymerase chain reaction analysis of genomic DNA failed to demonstrate a detectable difference between MS and control PBMC. PMID- 10814801 TI - Association of two variants in IL-1beta and IL-1 receptor antagonist genes with multiple sclerosis. AB - We studied the putative association of a C-->T polymorphism in exon-5 of IL-1beta and an 85 bp tandem repeat in intron-4 of IL-1 receptor antagonist (IL-1ra) genes with susceptibility to or outcome of MS. DNA from 122 cases from a population based cohort in Olmsted County, MN who were previously categorized for disease severity and temporal course and 244 ethnically-matched controls were analyzed. There was no association between either polymorphism and disease susceptibility. Allele-2 of IL-1beta and allele-3 of the IL-1ra polymorphisms were associated with a favorable outcome (P=0.023 and P=0.030). PMID- 10814802 TI - Hg(2+) and Cu(2+) interfere with agonist-mediated Ca(2+) signaling in isolated Mytilus digestive gland cells. AB - The effects of mercury and copper on agonist-mediated Ca-signaling were investigated in isolated cells from the marine mussel, Mytilus galloprovincialis Lam., by single cell fluorescence microscopy. In isolated digestive gland cells, short-term exposure (10 min) to both Hg(2+), a highly toxic metal and Cu(2+), an essential metal, in the nano-low uM range caused a sustained increase in cytosolic [Ca(2+)]. The effect of mercury on resting [Ca(2+)] was stronger than that of copper. The Hg-induced elevation in [Ca(2+)] seemed to be mainly due to an increased influx through Verapamil-sensitive Ca-channels, whereas the effect of Cu(2+) was related to a release from thapsigargin-sensitive intracellular stores. Agonists, such as epidermal growth factor (EGF), bradykinin (BK) and ATP, evoked Ca(2+) transients in isolated digestive gland cells through different mechanisms similar to those observed in mammalian cells, demonstrating the presence of common pathways of Ca-mediated cell signaling in both invertebrates and vertebrates. The agonist-mediated Ca(2+) response was affected by exposure to Hg(2+) and Cu(2+) in a concentration dependent manner: both metals significantly reduced the amplitude of the Ca(2+) spikes elicited by BK and ATP and decreased the percentage of EGF-responsive cells. The effects of Hg(2+) and Cu(2+) were apparently independent of their different type of interaction with the mechanisms involved in Ca(2+) homeostasis. The results clearly demonstrate that, in marine invertebrate cells, short-term exposure to heavy metal concentrations comparable to environmental exposure levels results in alterations of intracellular Ca(2+) homeostasis which compromise the cell response to extracellular stimuli involving Ca-mediated signaling. The mechanisms of heavy metal interference with Ca homeostasis and signaling are discussed. PMID- 10814803 TI - Total oxidant scavenging capacity (TOSC) of microsomal and cytosolic fractions from Antarctic, Arctic and Mediterranean scallops: differentiation between three potent oxidants. AB - The enhanced formation of reactive oxygen species (ROS) is a common pathway of toxicity induced by stressful environmental conditions. In polar environments, characterization of antioxidant defences in key sentinel species may be of particular value as early detection biomarkers of unforeseen effects of human activities which are progressively increasing in these remote areas.The complexities associated with predicting the consequences at the 'organism level' of variations of specific antioxidant defences have been recently overcome by the ability to quantify an index of specific biological resistance to various kinds of ROS.The total oxyradical scavenging capacity (TOSC) assay has been used in three species of scallops for quantifying their ability to neutralize peroxyl (ROO(&z.rad;)) and hydroxyl (&z.rad;OH) radicals and peroxynitrite (HOONO). Adamussium colbecki and Chlamys islandicus represent key organisms for monitoring Antarctic and Arctic regions while Pecten jacobaeus was chosen for a comparison with a related temperate species. TOSC values for ROO&z.rad; were significantly higher in A. colbecki indicating this species as the most efficient scavenger of ROO&z.rad;. Mediterranean scallops had the lowest TOSC for ROO(&z.rad;). A. colbecki also exhibited the highest scavenging capacity for &z.rad;OH with values more than 2-fold greater than for C. islandicus and P. jacobaeus. TOSC for HOONO was lower for all scallops as compared to those for ROO&z.rad; or &z.rad;OH. TOSC for microsomes was not significantly different among the species for any ROS studied, and the percentage contribution to the specific TOSC for the various oxidants of microsomes of all scallops accounted for 1-3% of the total TOSC of the post-mitochondrial fraction. The specific TOSC of scallop microsomes for &z.rad;OH was approximately ten times lower than that for ROO&z.rad; or HOONO.The higher basal capability of the Antarctic scallop to neutralize different reactive oxygen species is discussed in terms of a possible adaptation to this extreme environment and TOSC is validated as a quantifiable measure of susceptibility to oxidative stress in marine organisms. PMID- 10814804 TI - Activities of affinity-isolated glutathione S-transferase (GST) from channel catfish whole intestine. AB - A glutathione S-transferase (GST) fraction was isolated from cytosol prepared from catfish intestinal mucosa by GSH-agarose affinity chromatography and its molecular weight, isoelectric points, substrate specificities and immunochemical cross-reactivity were examined. Intestinal GSTs were purified 100-fold with respect to cytosolic activity with 1-chloro-2, 4-dinitrobenzene and had high activity with ethacrynic acid, (+/-)benzo(a)pyrene-4,5-oxide, and (+/-)anti benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide, but a low activity with 1,2-dichloro 4-nitrobenzene. SDS-polyacrylamide gel electrophoresis revealed the presence of a single band with relative molecular mass of 26700. Gel isoelectric focusing showed a major band with a pI of 8.2. A polyclonal antibody prepared against a GST pi-protein isolated from catfish proximal intestine cross-reacted well with the affinity isolated GST fraction. The catfish antibody also cross-reacted with GST from human placenta which contains predominantly pi-class GST (Mannervik, B., Guthenberg, C., 1981. Glutathione transferase (human placenta). In: Jakoby, W.B. (Ed.), Methods in Enzymology, 77. Academic Press, New York, pp. 231-235; Polidoro, G., Dillio, C., Arduini, A., Frederici, G., 1981. Molecular and catalytic properties of purified glutathione transferase from human placenta. Biochem. Med. 22, 247-259; Dao, D.D., Partridge, C.A., Kurosky, A., Awasthi, Y.C., 1982. Subunit structure of glutathione-S-transferase of human liver and placenta. IRSC Med. Sci, Lib. Compend. 10, 175; Dao, D.D., Partridge, C.A., Kurosky, A., Awasthi, Y.C., 1984. Human glutathione transferase. Characterization of the anionic forms from lung and placenta. Biochem. J. 221, 33-41), but poorly with human liver cytosol. The affinity-isolated protein fraction from whole intestine contained proteins that were immunologically related to all four major classes of human GSTs tested. N-terminal sequence analysis of the predominant band obtained by 2D electrophoresis indicated a marked homology (63-70% identical) to mammalian pi form GST isozymes and very strong similarity (80%) to a salmon hepatic GST that was designated a pi form (Dominey, R.J., Nimmo, I.A., Cronshaw, A.D., Hayes, J.D., 1991. The major glutathione S-transferase in salmonid fish livers is homologous to the mammalian pi class GST. Comp. Biochem. Physiol. (B) 100 (1), 93-98). Other bands contained insufficient protein for N terminal analysis. Taken together, these results indicate that the predominant intestinal GST isoform is related to the pi-class enzymes, but minor GSTs related to other families are also present. PMID- 10814805 TI - Effects of tetradifon on Daphnia magna during chronic exposure and alterations in the toxicity to generations pre-exposed to the pesticide. AB - Daphnia magna was exposed to some nominal sublethal tetradifon concentrations (0.10, 0.18, 0.22 and 0.44 mg/l) during 21 days. Chronic toxicity tests were carried out using neonates of F(1) first brood (F(1)-1st) and F(1) third brood (F(1)-3rd) offspring generations from parentals (F(0)) pre-exposed to the pesticide. The effect of tetradifon on survival, reproduction and growth was monitored for the selected daphnid generations. The parameters used to evaluate pesticide effect on reproduction were: mean total young per female, mean brood size, time to first reproduction, mean number broods per female and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to the pesticide. Reproduction was significantly reduced when tetradifon concentration increased in the medium. This effect was higher in F(1)-1st and F(1)-3rd offsprings compared to the parental generation (F(0)) daphnids. The intrinsic rate of natural increase (r) decreased with increasing concentrations of tetradifon especially in those animals from F(1)-3rd. However, the growth of the exposed organisms decreased in the same order of magnitud in all the generations tested. Survival was not affected after exposure to the selected tetradifon concentrations. The maximum acceptable toxicant concentration (MATC) was calculated for F(0), F(1)-1st and F(1)-3rd generations exposed to the pesticide using as parameter of evaluation the intrinsic rate of natural increase (r). The interpolation of these results gave MATC values of 0.13 mg/l pesticide for F(0) and F(1). The EC50 values have been derived for some selected parameters on D. magna exposed to tetradifon. EC50 values decreased in F(1)-1st and F(1)-3rd generations compared to the parental generation F(0), these results mean that less amount of toxicant would be necessary to reduce the selected parameters to 50% in the first and third broods of D. magna. Expanding the reproduction tests to several broods of a parental generation revealed important information on chronic toxicity that could be used in the protection of aquatic environment. PMID- 10814806 TI - The single substance and mixture toxicity of quinolones to the bioluminescent bacterium Vibrio fischeri. AB - Quinolones are one of the most important group of synthetic antibiotics used in aquaculture. We studied the single substance and mixture toxicity of ten quinolones using a long term bioluminescence inhibition assay with the marine bacterium Vibrio fischeri as the test organism. All tested quinolones are highly toxic to the test organism with EC50 values ranging from 14 ug/l for ofloxacin to 1020 ug/l for pipemidic acid. Adapting the approach outlined in EEC directive 93/21/EEC to these results, all but one of the ten quinolones belong to the group classified as being 'very toxic to aquatic organisms' (EC50 below 1 mg/l). On the basis of the concentration-response relationships of the single compounds, the mixture toxicity of the ten compounds was estimated by the concepts of concentration addition and independent action. Complete concentration-response relationships were experimentally recorded for the quinolone mixture in three different mixture ratios, based on the relative toxicity of the components (EC50, EC1 and NOEC). The results show that the mixture toxicity of the quinolones is best predictable by concentration addition, whereas independent action underestimates the toxicity of the mixture. As the quinolones have an identical specific mechanism of action (the inhibition of bacterial gyrases), these results are in agreement with the pharmacological assumptions that form the basis of the concept of concentration addition. It is therefore concluded, that concentration addition can be useful for hazard assessment procedures of mixtures of similarly acting compounds. One important implication of this concept is that even mixture components that are present only at their individual no observed effect concentrations (NOECs) contribute to the overall toxicity of the mixture. Under these conditions more than 99% effect of the quinolone mixture are observed. This result emphasises the unsuitability of NOECs as an approximation of a 'safe' concentration. PMID- 10814807 TI - Photoenhanced toxicity of weathered oil to Mysidopsis bahia. AB - The toxicity of a water-accommodated fraction (WAF) prepared from weathered oil was assessed in a 7-day static renewal test with Mysidopsis bahia. Weathered oil was collected from the 5x monitoring well at the Guadalupe oil field. Solar ultraviolet and visible light intensities were measured in various habitats in the vicinity of the weathered oil sample collection site, and the resultant measurements were used to produce laboratory light treatments that were representative of the on-site quality and intensity of natural solar radiation. Each of five WAF dilutions and a control without WAF was tested under three different simulated solar radiation intensities. During the test, survival and growth of the mysids, irradiance, and total petroleum hydrocarbon (TPH) concentrations in the test treatments were measured. Significant increases (Pgills>/=intestines>white muscle) and the gills of elasmobranchs (order: gills>liver>white muscle>intestines). Rainbow trout accumulated more silver than the other teleosts, and were the only species to suffer significant mortality, effects likely associated with added salinity stress. Accumulations were fairly uniform amongst the other teleosts. Similar concentrations in gills and intestines suggested that both branchial and intestinal uptake occurred, with the latter potentially dominant; indeed sole exhibited no silver build-up in the gills. The two elasmobranchs exhibited no silver build-up in intestines but much higher levels in gills, indicating that in the absence of drinking, only branchial uptake occurs. Nevertheless, based on whole liver content, the elasmobranchs accumulated silver 5-15-fold faster than the three teleosts. Over 21-day exposures (1.5-50.0 ug/l Ag) in sculpin, salinity markedly affected silver accumulation, with tissue-specific levels approximately 6-fold higher at 18, than at 30 ppt. However, there was negligible effect of salinity on silver accumulation during 48 h at 250 ug/l Ag. Silver bioaccumulation appears to be markedly affected by speciation. At lower salinities, or higher [Ag], a neutral charged AgCl(aq) complex exists in the water, allowing for increased bioaccumulation to occur. At higher salinity, only less bioavailable, negatively charged AgCl(n)(1-n) complexes are present (AgCl(2)(-), AgCl(3)(2-), AgCl(4)(3 )). PMID- 10814812 TI - Effects of contaminants in the Antarctic environment - potential of the gammarid amphipod crustacean Paramorea walkeri as a biological indicator for Antarctic ecosystems based on toxicity and bioacccumulation of copper and cadmium. AB - This study provides information on LC(50) toxicity tests and bioaccumulation of heavy metals in the nearshore Antarctic gammarid, Paramorea walkeri. The 4 day LC(50) values were 970 ug/l for copper and 670 ug/l for cadmium. Net uptake rates and bioconcentration factors of these elements were determined under laboratory conditions. After 12 days of exposure to 30 ug/l, the net uptake rates were 5.2 and 0.78 ug/g per day and the bioconcentration factors were 2080 and 311 for copper and cadmium, respectively. The body concentrations of copper were significantly correlated with the concentrations of this element in the water. Accumulation of copper and cadmium continued for the entire exposure suggesting that heavy metals concentrations were not regulated to constant concentrations in the body. Using literature data about two compartments (water-animal) first-order kinetic models, a very good agreement was found between body concentrations observed after exposure and model predicted. Exposure of P. walkeri to mixtures of copper and cadmium showed that accumulation of these elements can be assessed by addition of results obtained from single exposure, with only a small degree of uncertainty. The study provides information on the sensitivity of one Antarctic species towards contaminants, and the results were compared with data of similar species from lower latitudes. An important finding is that sensitivity to toxic chemicals and toxicokinetic parameters in the species investigated are comparable with those of non-polar species. The characteristics of bioaccumulation demonstrate that P. walkeri is a circumpolar species with the potential to be a standard biological indicator for use in monitoring programmes of Antarctic nearshore ecosystems. The use of model prediction provide further support to utilise these organisms for biomonitoring. PMID- 10814813 TI - Effects of pH on cadmium and zinc uptake by the midge larvae Chironomus riparius. AB - We studied the effect of pH on the uptake of cadmium and zinc by fourth instar larvae of the midge Chironomus riparius within the pH range 5.5-10.0, using chemically defined solutions. The effect of prior acclimation on metal uptake was examined for four pH levels, i.e. pH 5.5, 7.0, 8.0 and 9.5. At least three factors were important in determining the effect of pH on the cadmium and zinc uptake by midge larvae. The effect of pH on metal uptake is the combined result of changes in free metal ion activity, changes in pH of exposure and changes in pH of acclimation, the latter representing a physiological effect. Within each acclimation group metal uptake in larvae increased with increasing pH of exposure in the range 5.5-9.0 but decreased between pH 9.0 and 10.0. Taking into account the decreased free metal ion activity, metal uptake was still high at pH 10.0. A possible explanation for this is that an increase in pH alters the metal uptake process by decreasing the protonation of the binding sites. That is, the biological availability of the free metal ion increases with increasing pH. Among the different pH exposure groups, acclimation had a positive effect up to pH 9.0 but a negative effect between 9.0 and 10.0. Two different uptake models were applied to describe the observed variation in metal uptake. With a non-linear, semi-empirical model, the integration of the different pH effects for the pooled data described no more than 38% of the total variation in cadmium uptake and 36% of the total variation in zinc uptake by midge larvae. When the model was fitted to the uptake data of larvae acclimated to the exposure conditions, 78 and 69% respectively of the variation was described. The second model, a biological ligand model, was not able to discriminate between effects of pH in acclimated and non-acclimated exposure groups. Only for the data of larvae acclimated to the exposure conditions the model could describe a significant amount of the observed variation in metal uptake, R(2) values being comparable to those of the first model. The remaining high undescribed variation could be ascribed to the high natural variation in metal uptake by midge larvae. PMID- 10814814 TI - Characterization and transcriptional profiles of two rice MADS-box genes. AB - The plant MADS-box gene family plays a key role in plant development, especially in flower development. We designed degenerate primer according to the MADS-box conserved region and isolated two cDNA from rice, FDRMADS6 and FDRMADS7, which are homologous to AP1. RT-PCR expression analyses by using total RNA isolated from root, shoot and flower showed that the FDRMADS6 transcript was detectable only in flower while FDRMADS7 was expressed in all three tissues. In situ hybridization experiments indicated that at the early stage of rice flower development, the transcripts of FDRMADS6 and FDRMADS7 were detected in the spikelet apical meristem, which were same as AP1. At the late stage, when flower organ primordia started differentiating, the expression of FDRMADS6 appeared to be specifically localized in developing stamens and the pistil primordia, while the transcripts of FDRMADS7 were detectable abundantly throughout the organ primordia. Our results suggest the two MADS-box genes may be members of the AP1 family, but may have different functions. PMID- 10814815 TI - Isolation and characterisation of the carnation floral-specific MADS box gene, CMB2. AB - The cDNA clone KD81, was isolated from a carnation petal cDNA library based on its strong differential expression in petals compared with leaves. The deduced amino acid sequence of KD81 indicated high homology with members of the MADS box family of transcription factors. Identified within the deduced amino acid sequence are two conserved domains; an N-terminal, MADS box and a central, K box. The gene encoding KD81 was termed Carnation MADS Box gene 2 (CMB2). Expression of CMB2 is floral-specific and in petal, transcripts were persistent from the initial stages of development through flower opening. Transcripts were not detected in vegetative tissues. The CMB2 protein is most homologous to TDR6 from tomato, the product of the petal and stamen identity gene DEFICIENS (DEFA), and several DEFA homologues including SLM3, STDEF, PMADS1 and APETALA3. Southern blot analysis indicated that CMB2 is present as a single copy within the carnation genome. Characterisation of a genomic clone encoding CMB2, revealed the molecular structure of CMB2 to be consistent with that reported for other plant MADS box genes. Analysis of the CMB2 promoter sequence revealed the presence of two putative cis-acting elements known as serum response elements (SREs). These elements are proposed as the target for MADS box domain binding and may be involved in the regulation/autoregulation of gene expression. CMB2 represents the first reported isolation of a MADS box gene from carnation. PMID- 10814816 TI - An efficient in vitro procedure for micropropagation and generation of somaclones of rose scented Pelargonium. AB - Efficient protocols have been established for both direct and indirect regeneration of plants in Pelargonium graveolens Indian cultivar Hemanti (Algerian type). Murashige and Skoog's (MS) medium [T. Murashige, F. Skoog, A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15 (1962) 473-497] supplemented with 5.0 mg/l kinetin and 1.0 mg/l NAA was optimal for direct regeneration of plants from leaf explants while 8.0 mg/l kinetin and 1.0 mg/l NAA proved optimum for nodal explants for maximum number of shoots per explant. Callus induction was observed from nodal explants on MS medium supplemented with 10 mg/l kinetin and 1.0 mg/l NAA. Callus on further transfer to MS medium with 0.5 mg/l BAP and 0.1 mg/l NAA exhibited regeneration of maximum number of shoots. In vitro grown shoots of both direct and indirect origin rooted within 7-10 days following transfer to half strength MS medium with 1.0 mg/l IBA. Plantlets were acclimatized under glass house conditions with 90% survival. Randomly selected 85 individual Calliclones were subjected to field trial with 85-95% survival for two successive years along with control in randomized block design with three replicates. Screening of these calliclones revealed two distinct morphotypes, one with parental type highly dentated leaves (HDL) and the other with less dentated, round leaves (LDL). Only HDL calliclones flowered under field conditions. The LDL clones differed in several herb related agronomic characteristics such as plant height, herb yield, canopy size and number of branches per plant from the parental type as well as from the parent, which seems advantageous for commercial exploitation of such clones. The HDL clones closely resemble the parent in having higher content of citronellol than geraniol while the LDL clones contain almost equal contents of citronellol and geraniol in their essential oils as revealed by gas chromatography analysis. It is noticeable that the variability both in terms of agronomic characters and essential oil profiles among the clones were stable over 2 years of field trials. PMID- 10814817 TI - Functional analysis of a nitrite reductase promoter from birch in transgenic tobacco. AB - Nitrate assimilation is a highly regulated process in higher plants, and the regulatory cues governing gene expression in this pathway include both external and internal factors. In birch (Betula pendula Roth) the expression of nitrate reductase (NR) and nitrite reductase (NiR) genes is co-regulated by light and nitrate at the transcriptional level. In order to identify cis-acting DNA elements involved in light and nitrate induction of the birch NiR gene, a 0.9 kb 5' flanking region of the NiR gene was isolated, analysed on the DNA level, and the transcription start site was determined. Deletion analysis of the birch NiR promoter region fused to the GUS reporter gene (uidA) in transgenic tobacco (Nicotiana tabacum) revealed the presence of light- and nitrate-responsive promoter fragments. The responsive fragments showed different activities in leaves and roots. Further, gel mobility shift assays using nuclear proteins from leaves detected a specific DNA-binding activity to the sequence between -146 and 267 bp that was induced in darkness and disappeared in the light. The deletion analysis has shown that this region is critical for light inducibility of the birch NiR gene in leaves. PMID- 10814818 TI - Possible involvement of protein phosphorylation in the wound-responsive expression of Arabidopsis plastid omega-3 fatty acid desaturase gene. AB - The plastid omega-3 fatty acid desaturase (FAD7) catalyzes the conversion of linoleic acid to linolenic acid. Wounding enhances the expression of the FAD7 gene in leaves and induces its expression in stems and roots. The wound-induced expression of the FAD7 promoter was investigated in transgenic tobacco plants carrying the -825 Arabidopsis FAD7 promoter::beta-glucuronidase (GUS) fusion gene. The protein kinase inhibitor, staurosporine, and the protein phosphatase inhibitor, calyculin A, suppressed the wound induction of the FAD7 gene in stems. A tobacco mitogen-activated protein kinase (WIPK) was rapidly activated upon wounding not only in leaves but also in stems and roots, indicating that WIPK probably mediates the wound signals in most vegetative organs. The FAD7 promoter::GUS fusion gene was introduced into the transgenic tobacco plants in which the wipk gene was expressed constitutively at a high level or into the transgenic plants in which the wipk gene was suppressed possibly due to the transgene-induced gene silencing. The wound-induced expression of the FAD7 gene in stems was enhanced in the former transgenic tobacco plants and suppressed in the latter plants. These results suggest that the wound activation of the FAD7 promoter depends on both protein phosphorylation and dephosphorylation events especially in stems, and also that WIPK is involved in such signaling cascades. PMID- 10814819 TI - Characterisation of microsatellite markers from sugarcane (Saccharum sp.), a highly polyploid species. AB - Cultivated sugarcane varieties (Saccharum spp) are derived from complex interspecific hybridisations between the species S. spontaneum (2n=40-128) and S. officinarum (2n=60 or 80). To analyse this complex genome, the potential of microsatellite repeats as genetic markers in sugarcane with respect to their abundance, variability and ability to detect polymorphisms was investigated. A set of microsatellite markers for genome analysis in cultivated sugarcane was identified from an enriched genomic DNA library constructed from Saccharum sp. cv Q124. Sequencing of 798 sugarcane genomic DNA clones from an enriched microsatellite library, yielded 457 inserts containing microsatellite repeat motifs. Just over 84% of the microsatellites contained dinucleotide or trinucleotide repeats averaging 15 and 13 repeat motifs, respectively. Primer sets were designed and synthesised for over 100 microsatellite sequences and tested on a set of five sugarcane cultivars. Both, heterozygosity as witnessed by the number of alleles, and length polymorphisms as seen in the differences in PCR product size for a particular allele were observed. Microsatellite markers are likely to have many applications in sugarcane genetics and breeding including germplasm analysis, cultivar identification, parent evaluation and marker assisted breeding. PMID- 10814820 TI - Genomic organization and expression properties of the VfENOD5 gene from broad bean (Vicia faba L.). AB - A full-length cDNA encoding the broad bean (Vicia faba L.) early nodulin VfENOD5 was isolated from a nodule cDNA library. In addition to the ENOD5 homologues from other legumes the derived VfENOD5 amino acid sequence also displayed homologies to the phytocyanin-related nodulins GmENOD55-2, MtENOD16, and MtENOD20. A close inspection of the ENOD5 proteins from broad bean, pea and vetch indicated that all these nodulins possess a putative C-terminal GPI-anchor signal sequence. This novel finding supports the hypothesis that ENOD5 is an arabinogalactan protein. Tissue print hybridizations revealed that the broad bean ENOD5 gene was not only expressed in the central tissues of root nodules. In contrast to other legumes hybridizing transcripts were also be detected in a narrow zone within the peripheral nodule tissues. Sequence analysis of a genomic clone indicated the presence of a single intron interrupting the VfENOD5 coding region at a position precisely corresponding to the MtENOD16 and MtENOD20 introns. PMID- 10814821 TI - Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation. AB - An Agrobacterium tumefaciens-mediated transformation system was developed for Artemisia annua L. Using this system a cDNA encoding farnesyl diphosphate synthase (FDS placed under a CaMV 35S promoter) was transferred into A. annua via A. tumefaciens strain LB4404. Leaf or leaf discs were used as explants to be infected with A. tumefaciens and an optimal concentration of 20 mg/l kanamycin was applied to select kanamycin resistant shoots. Forty-five lines of resistance kanamycin shoots transformed with FDS were established. Analysis of PCR showed that at least 20 shoots transformed with the FDS gene were PCR positive. Southern blot analysis suggested the foreign FDS gene had been integrated into the A. annua genome, and Northern blot analysis revealed that the foreign FDS gene expressed at the transcriptional level in five shoot lines (F-1, F-4, F-61, F-62 and F-73 shoot lines). Analysis of artemisinin demonstrated that about 8 approximately 10 mg/g DW of artemisinin were then detected in transgenic plants regenerated from five shoot lines, this is about 2-3 times higher than that in the control. PMID- 10814822 TI - Plastid gene expression is not associated with midday depression in CO(2) assimilation and electron transport. AB - To investigate the effect of diurnal variations on chloroplastic electron transport as well as accumulation of gene products associated with it, studies were carried out on Populus deltoides, a tree species. Electron transport studies showed two peak responses as a large diurnal change with pronounced midday depression in whole chain (H(2)O--> MV) as well as partial reactions for PSII (H(2)O-->PBQ) and PSI (DCPIP-->MV). The electron transport rates first increased from 05:00 h to a maximum at around 09:00 h and then showed a decrease at 13:00 h followed by recovery and further decrease. The pigments associated with electron transport chain did not show any change during the day. Surprisingly midday depression in the accumulation of transcripts and polypeptides related to electron transport was not observed. This suggests that chloroplastic gene expression is not associated with the midday depression observed for both CO(2) assimilation and electron transport. Studies on the transcripts of psbD/C operon during the day showed that there were differences in the processing pattern although the steady state levels of the processed transcripts of this operon did not show any variation. PMID- 10814823 TI - Targeting of the soybean leghemoglobin to tobacco chloroplasts: effects on aerobic metabolism in transgenic plants. AB - Several attempts have been made to alter the aerobic metabolism of plants, especially those related to the oxygenation or carboxylation of Rubisco. However, designing a more efficient Rubisco protein is rather problematic since its structural manipulation leads frequently to an enhancement of oxygenase activity, which is responsible for photorespiratory losses. In order to reduce oxygen availability inside the chloroplast, a chimeric gene consisting of a soybean leghemoglobin cDNA (lba) ligated to the chloroplast targeting signal sequence of the Rubisco small subunit gene, was introduced and expressed in Nicotiana tabacum. Lb was efficiently imported and correctly processed inside the chloroplasts of transgenic tobacco plants. Furthermore, the level of Lb expression in leaf tissue ranged from 0.01 to 0.1%. Analysis of photosynthesis, starch, sucrose and enzymes involved in aerobic metabolism, revealed that despite the high affinity of Lb for oxygen, no significant difference was observed in relation to the control plants. These results suggest that higher Lb concentrations would be required inside the chloroplasts in order to interfere on aerobic metabolism. PMID- 10814824 TI - GFLV replication in electroporated grapevine protoplasts. AB - Grapevine fanleaf virus (GFLV), responsible for the economically important court noue disease, is exclusively transmitted to its natural host in the vineyards through Xiphinema nematodes. We have developed direct inoculation of GFLV into grapevine through protoplast electroporation. Protoplasts were isolated from mesophyll of in vitro-grown plants and from embryogenic cell suspensions. Permeation conditions were determined by monitoring calcein uptake. Low salt poration medium was selected. Electrical conditions leading to strong transient gene expression were also tested for GFLV inoculation (isolate F13). GFLV replication was detected with either virus particles (2 ug) or viral RNA (10 ng) in both protoplast populations, as shown by anti-P38 Western blotting. Direct inoculation and replication were also observed with Arabis mosaic virus (ArMV), a closely related nepovirus, as well as with another GFLV isolate. These results will be valuable in grapevine biotechnology, for GFLV replication studies, transgenic plant screening for GFLV resistance, and biorisk evaluation. PMID- 10814825 TI - Naringenin 7-O-methyltransferase involved in the biosynthesis of the flavanone phytoalexin sakuranetin from rice (Oryza sativa L.). AB - An inducible S-adenosyl-L-methionine:naringenin 7-O-methyltransferase (NOMT) catalyzing the methylation of naringenin to sakuranetin, a major rice phytoalexin was purified approximately 985-fold from ultraviolet (UV)-irradiated rice leaves. The enzyme is not found in healthy tissues and was purified to a nearly homogeneous preparation in one step using adenosine-agarose affinity chromatography, with 1 g rice leaves (UV-irradiated) as starting material. Gel filtration chromatography resulted in an almost pure enzyme, as evidenced by a major band migrating to a position corresponding to a molecular mass of approximately 41 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified NOMT was strongly inhibited by Mn(2+), Ni(2+), Cu(2+), Zn(2+), Hg(2+), and Cd(2+), and to a low degree by Co(2+), Mg(2+), Ba(2+), Ca(2+) and ethylenediamine tetraacetic acid. The amino acid sequence of a NOMT cyanogen bromide (CNBr)-cleavage peptide was highly homologous to that of a caffeic acid 3-O-methyltransferase from maize, and about 70% of the amino acid sequence was obtained after sequencing the peptides generated by CNBr and/or formic acid hydrolysis. NOMT was also shown to be induced in a time-dependent manner, and purified from rice leaves treated with jasmonic acid and copper chloride. PMID- 10814826 TI - An alternatively spliced transcript of the rat nociceptin receptor ORL1 gene encodes a truncated receptor. AB - Opioid receptor-like protein ORL1, the receptor for the neuropeptide nociceptin (also named orphanin FQ), has two alternatively spliced isoforms in the rat. This alternative splicing event is generated by retaining of intron 3, 81 bases in length, in the mRNA region encoding the second extracellular loop of ORL1. A full length rat ORL1 receptor has 367 amino acid residues. However, as revealed by sequencing of rat ORL1 genomic DNA and cDNA, the insertion of the unspliced intron 3 brings in an in-frame stop codon and, therefore, creates a truncated open-reading frame encoding only the N-terminal half of ORL1 (from the N-terminus to an alternate extracellular tail C-terminal to the fourth transmembrane domain). The two alternatively spliced transcripts are differentially expressed in tissues. In transfected mammalian cells, the full-length ORL1 displays high affinity and selective binding for nociceptin, and inhibits the production of cyclic AMP. In contrast, the truncated ORL1 binds nociceptin and other opioid peptides very poorly and non-selectively (affinity in micromolar range), and it does not mediate any inhibitory effects on cyclic AMP production. Apparently, this truncated ORL1 does not function as a receptor for nociceptin or other ligands tested. Such alternative splicing to create a truncated ORL1 receptor might be an endogenous mechanism to negatively regulate nociceptin/ORL1 functions. PMID- 10814827 TI - Gene expression of taurine transporter and taurine biosynthetic enzymes in brain of rats with acute or chronic hyperosmotic plasma. A comparative study with gene expression of myo-inositol transporter, betaine transporter and sorbitol biosynthetic enzyme. AB - Cells exposed to hyperosmotic conditions maintain their volume by accumulating organic osmolytes. Taurine is considered as an osmolyte in brain cells. Accumulation of other osmolytes (sorbitol, myo-inositol and betaine), was shown in renal cells to result from an upregulation of the expression of the genes regulating osmolyte cell content. We have investigated the gene expression of the taurine transporter (TauT) and of the taurine biosynthetic enzymes, cysteine dioxygenase (CDO) and cysteine sulfinate decarboxylase (CSD) by measuring their mRNA levels in brain of salt-loaded rats. mRNA levels of genes previously identified as osmosensitive, namely aldose reductase (AR), myo-inositol transporter (SMIT) and betaine transporter (BGT1) were also determined. In whole brain, TauT-, SMIT- and BGT1-mRNA levels were significantly increased following acute salt-loading but SMIT-mRNA levels only remained elevated following chronic salt-loading while CDO-, CSD- and AR-mRNA levels remained unchanged in both conditions. Following acute salt-loading, mRNA levels of TauT, CDO, CSD, SMIT, BGT1 and AR were increased in cerebral cortex while SMIT- and BGT1-mRNA levels only were increased in striatum and habenula.TauT, CDO and CSD genes may be upregulated in brain of salt-loaded rats but the upregulation of the TauT gene appears more widespread. TauT, CDO and CSD are thus putative osmosensitive genes. However the actual pattern (amplitude, time course and regional occurrence) of the upregulation of each of the putative (TauT, CDO and CSD) and established (AR, SMIT and BGT1) osmosensitive genes differs markedly. This indicates that there exist other factors in brain cells which can selectively prevent the upregulation of these genes by hyperosmolarity. PMID- 10814829 TI - Selective regional blockade of junB gene expression in the hamster suprachiasmatic nucleus by a tyrosine kinase inhibitor. AB - The hypothalamic suprachiasmatic nucleus (SCN) functions as a circadian pacemaker regulating a variety of physiological and behavioral rhythms in mammals. Retinal illumination evokes expression of several immediate-early genes, including junB, in the ventral SCN early in the subjective night and throughout the SCN later in the subjective night. junB mRNA and protein are also expressed spontaneously around subjective dawn in nocturnal rodents, but only in the dorsal SCN. We examined the biochemical signaling mechanisms underlying both spontaneous and light-evoked expression of junB mRNA in the SCN of Syrian hamsters. Hamsters were injected (i.p.) before subjective dawn with vehicle or with either tyrphostin or genistein, inhibitors of protein tyrosine kinase, and maintained in the dark for 30 min. They were then exposed to a light pulse or kept in darkness for another 30 min. In situ hybridization studies demonstrated that tyrphostin pretreatment (12 or 24 mg/kg) reduced both spontaneous and light-evoked expression of junB mRNA only in the dorsal, and not the ventral, portion of the SCN. Conversely, genistein had little effect on either spontaneous or light-evoked expression of junB mRNA in any part of the SCN. These results indicate that a protein tyrosine kinase sensitive to tyrphostin but not to genistein is involved in the transduction pathways leading to expression of junB mRNA selectively in the dorsal SCN, independently of circadian phase and independently of the involvement of light. PMID- 10814828 TI - The coxsackievirus-adenovirus receptor protein as a cell adhesion molecule in the developing mouse brain. AB - In an attempt to elucidate the molecular mechanisms underlying neuro-network formation in the developing brain, we analyzed 130 proteolytic cleavage peptides of membrane proteins purified from newborn mouse brains. We describe here the characterization of a membrane protein with an apparent molecular mass of 46 kDa, a member of the immunoglobulin superfamily of which the cDNA sequence was recently reported, encoding the mouse homologue of the human coxsackievirus and adenovirus receptor (mCAR). Western and Northern blot analyses demonstrated the abundant expression of mCAR in the mouse brain, the highest level being observed in the newborn mouse brain, and its expression was detected in embryos as early as at 10. 5 days post-coitus (dpc), but decreased rapidly after birth. On in situ hybridization, mCAR mRNA expression was observed throughout the newborn mouse brain. In primary neurons from the hippocampi of mouse embryos the expression of mCAR was observed throughout the cells including those in growth cones on immunohistochemistry. In order to determine whether or not mCAR is involved in cell adhesion, aggregation assays were carried out. C6 cells transfected with mCAR cDNA aggregated homophilically, which was inhibited by specific antibodies against the extracellular domain of mCAR. In addition to its action as a virus receptor, mCAR may function naturally as an adhesion molecule involved in neuro network formation in the developing nervous system. PMID- 10814830 TI - Structure of the mouse NDRF gene and its regulation during neuronal differentiation of P19 cells. AB - We have isolated and characterized the mouse gene for NDRF (neuroD-related factor), a basic helix-loop-helix transcription factor implicated in neural development and function. The gene consists of two exons and the entire protein coding sequence is encoded by a single downstream exon. RNA blot hybridization analysis revealed that NDRF mRNA was detectable at day 4 and increased to a maximal level at day 6 during neuronal differentiation of P19 cells. To elucidate the regulatory mechanisms of the NDRF gene expression during this process, a construct containing the genomic DNA fragment of about 3 kbp upstream of the NDRF coding region fused to a luciferase reporter gene was transfected into P19 cells, and stable transformants were pooled for assay of luciferase activities. When the stable transformants were treated with RA and aggregated to induce neuronal differentiation, the luciferase activities were induced in a temporal expression pattern similar to that of the endogenous NDRF mRNA. Further experiments using a series of deletion and mutation constructs indicated that the 376-bp sequence in the 5'-flanking region of the NDRF gene is important, and that one of the E boxes in the sequence plays a critical role in the regulated expression. Transient transfection experiments also showed that the same E box is required for the transactivation of the NDRF promoter activity by neurogenin 1. These results suggest that the NDRF gene expression is regulated by an E box-binding factor during neuronal differentiation of P19 cells. PMID- 10814831 TI - Puralpha, a single-stranded DNA binding protein, suppresses the enhancer activity of cAMP response element (CRE). AB - Puralpha, a single-stranded DNA binding protein, recognizes a PUR element (GGN repeat). We have reported that Puralpha binds to a single-stranded oligonucleotide probe containing the cAMP response element (CRE) of rat somatostatin gene using a gel mobility shift assay. Here, we showed that Puralpha binds to the probe only in the presence of a PUR element by a more detailed characterization. We also examined the effects of Puralpha on the enhancer activity of the somatostatin CRE in PC12 cells using the reporter gene assay. Transfected Puralpha suppressed the CRE enhancer activity stimulated by forskolin (which increases intracellular cAMP), but suppression was not observed when the PUR element was deleted. The neurite extension induced by forskolin was inhibited by the transfection of Puralpha, but that by NGF was not suppressed. The c-fos mRNA induced by forskolin, but not by NGF, was also suppressed by Puralpha transfection. These results indicate that Puralpha suppresses the biological activities induced by forskolin, but not by NGF, in PC12 cells and that Puralpha could interfere with a cAMP-CRE signal pathway. PMID- 10814832 TI - Prior experience of morphine application alters the c-fos response to MDMA ('ecstasy') and cocaine in the rat striatum. AB - Repeated morphine application usually leads to the development of tolerance but under certain circumstances sensitization may arise simultaneously. This phenomenon becomes obvious in behavioral tests as increasing locomotor activity and increasing drug self-administration during a course of chronic morphine application. It was suggested recently that sensitization could contribute to addiction. The molecular mechanisms of sensitization may include the long lasting increase in neuronal responsiveness to morphine which was observed in defined brain areas after repeated morphine injections. In this work, we studied whether morphine-sensitized Wistar rats also display an enhanced neuronal activity in response to other drugs of abuse (so called co-sensitization). The substances to be tested were injected as single doses 4 weeks after completion of a 10-day morphine pretreatment. MDMA (3, 4-methylenedioxymethamphetamine, 6 mg/kg) as a single test dose yielded a c-fos response in a wide range of brain areas. In the caudate putamen, the expression pattern of c-fos was clearly altered if the rats had received repeated morphine application previously. In this case, the MDMA induced c-fos expression was markedly confined to the centromedial, mesolimbic aspect of the striatum whereas it had a diffuse appearance in rats not exposed to the opiate earlier. Cocaine application (50 mg/kg) elicited an intense c-fos expression in the medial striatum if the animals were morphine-pretreated; it was virtually absent in drug-naive rats after the same cocaine dose. Ten mg/kg cocaine had a similar but weaker effect. No difference in the c-fos expression pattern between morphine and saline pretreated animals was observed in the case of a THC (Delta(9)-tetrahydrocannabinol, 25 mg/kg) or an LSD (lysergic acid diethylamide, 1 mg/kg) test application. These findings imply that morphine sensitizes the brain towards other addicting drugs. In consequence, morphine sensitization obviously does not solely reflect alterations in mu-opioid receptor signaling. Rather, it seems to reflect further rearrangements within the mesolimbic system. PMID- 10814833 TI - Differential activation of MAPK/ERK and p38/SAPK in neurones and glia following focal cerebral ischaemia in the rat. AB - Two relatively well characterised kinase signalling pathways are those involving MAPK/ERK and p38/SAPK2, that are known to be activated in vitro by various factors known to increase following stroke, such as glutamate, IL-1 and TNF. The present study was designed to investigate the activation and cellular distribution of phosphorylated-ERK1/2, -p38 and the transcription factor CREB following focal cerebral ischaemia using phosphospecific antibodies. Up to 24 h following transient MCAO (90 min) and 6 h following permanent MCAO, phospho ERK1/2 staining was markedly increased within the cytoplasm of neuronal perikarya in 'penumbral-like' regions. In contrast, phospho-p38 immunostaining was markedly increased in cells with astrocyte-like morphology in both 'core' and 'penumbral like' regions. Phospho-p38 staining was also detected in some neurones within 'penumbral-like' regions up to 24 h following transient MCAO. CREB activation was confined to neurones in 'penumbral-like' regions. Increased phospho-p38 immunoreactivity was detected in astrocyte-like cells present in the subcortical white matter ipsilateral to the occluded MCAO, while phospho-CREB and -ERK1/2 staining was localised to cells with the morphological appearance of oligodendrocytes. This study demonstrates phosphorylation, indicative of activation, of both the MAPK and p38 pathways following transient and permanent MCAO. However, each pathway shows a distinct cellular and spatial distribution within ischaemic tissue. Together these data indicate that neuroprotection offered by agents directed towards the ERK1/2 pathway may act directly through protection of neurones and oligodendrocytes, while those directed towards the p38 pathway kinase signalling pathways may be indirectly via inhibition of cytokines and other mediators involved in the brains response to injury. PMID- 10814834 TI - Immunohistochemical study of the distribution of Ca(2+)/calmodulin-dependent protein kinase phosphatase in the rat central nervous system. AB - Distribution of Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaM-K Pase) which dephosphorylate multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaM-kinases) in the rat brain and spinal cord were examined immunohistochemically by using an antibody against this enzyme. CaM-K Pase was localized only in the cytoplasm as has been investigated in PC 12 cells, and was never observed in the nucleus. Immunostainability varied from cell group to cell group. Mitral cells in the olfactory bulb, pyramidal neurons in the fifth layer of the cerebral cortex, hippocampal and striatal interneurons, dorsal and ventral pallidal, entopeduncular, and the reticular part of the substantia nigra neurons were intensely immunolabeled. Motoneurons in all the cranial nerve nuclei and the anterior horn of the spinal cord also revealed intense immunolabeling. On the contrary, pyramidal neurons in the Ammon's horn of the hippocampal formation, granule cells in the olfactory bulb, dentate gyrus and cerebellar cortex, Purkinje cells, neurons in the medial habenular nucleus and the inferior olivary nucleus have not shown immunoreactivity. Axons in the white matter or nerve root of the cranial nerve nuclei were immunolabeled. Glial cells in the white matter also showed immunostaining. Because the substrate of CaM-K Pase is multifunctional CaM-kinase II, I and IV, localization of each CaM-kinase was compared with that of CaM-K Pase. The distribution of CaM-K Pase and these CaM kinases was found to overlap in various regions in the brain and spinal cord. It was concluded, therefore, that CaM-K Pase could regulate the activity of these CaM-kinases by dephosphorylation, when they existed together in neurons. PMID- 10814835 TI - Expression of the protein tyrosine phosphatase, phosphatase of regenerating liver 1, in the outer segments of primate cone photoreceptors. AB - Foveal cone photoreceptors are morphologically distinct and, presumably, express unique transcripts. We have identified a cDNA clone encoding the protein tyrosine phosphatase (PTP), phosphatase of regenerating liver 1 (PRL-1) in a screen for genes that are enriched in monkey fovea. PRL-1 was originally isolated as an immediate early gene in regenerating liver [R.H. Diamond, D.E. Cressman, T.M. Laz, C.S. Abrams, R. Taub, PRL-1, a unique nuclear protein tyrosine phosphatase, affects cell growth, Mol. Cell Biol. 14 (1994) 3752-3762]. On cDNA Southern blots of human and monkey retina, radiolabeled PRL-1 cDNA hybridized to a single mRNA species of about 2.5 kb that was most intense in fovea-enriched samples. The monkey PRL-1 deduced amino acid sequence is identical to human, rat and mouse PRL 1. Affinity-purified antibodies directed against PRL-1 preferentially labeled cone photoreceptor cells and a subpopulation of bipolar cells in monkey retina. Immunoreactivity in cones was confined to red and green, but not to blue, cones and was restricted to the outer segments. Immunolocalization also revealed that PRL-1 protein expression was non-nuclear, suggesting that its function in the retina may be unrelated to its role in other tissues where it is expressed primarily in nuclei. Although both foveal and extrafoveal cones were PRL-1 reactive, the high abundance of PRL-1 mRNAs detected in monkey fovea correlates with the high concentration of cones in the fovea. The PRL-1 gene is located on chromosome 6q within an interval that also contains the genes that cause two hereditary retinal dystrophies. These studies demonstrate novel expression of the PRL-1 gene in the neural retina and suggest the phosphatase activity of PRL-1 may modulate normal cone photoreceptor cell function. PMID- 10814836 TI - Expression of the orphan receptor TR4 during brain development of the rat. AB - The orphan receptor TR4, member of the nuclear hormone receptor family, is related to the orphan receptors TR2, COUP-TFI and ARP-1, and was originally cloned from the adult rat brain. The latter two orphan receptors have been implicated in central nervous system (CNS) development. To investigate a possible role for TR4 in brain development, expression of TR4 was studied in rat embryos. At embryonic days 14.5 and 19.5, high expression of TR4 was found in the CNS, while low expression was detected throughout the embryo. In postnatal rats, TR4 was mainly expressed in the hippocampus and cerebellum, resembling the expression pattern found in adult brain. These data show that like COUP-TFI and ARP-1, expression of TR4 becomes restricted to distinct areas. In adult brain, TR4 is predominantly expressed in granule cells of both hippocampus and cerebellum. The data suggest a possible role for TR4 during proliferation and maturation of brain structures. PMID- 10814837 TI - GABA(B) receptor heterodimer-component localisation in human brain. AB - In recombinant cell lines, functional GABA(B) receptors are only formed by the heterodimerisation between two related G-protein coupled receptor proteins GABA(B)R1 (GBR1) and GABA(B)R2 (GBR2), whilst the individual GBR1 or GBR2 do not produce fully functional receptors. To determine whether the heterodimerisation occurs in vivo, novel polyclonal antibodies targeting the C termini of GBR1 and GBR2, were raised in different species, characterised, and used to determine the relative localisation of the reported heterodimer components in human brain tissue, using immunohistochemistry. The use of different species for the raising of the antisera allowed double immunofluorescent labelling of the receptors as an indication of GBR1/GBR2 receptor co-localisation in human brain. The presence of both proteins is reported in cerebellum, hippocampus, cortex, thalamus and basal ganglia. Regions of the brainstem including pons and medulla, also express GBR1 and GBR2 protein. The double immunofluorescence demonstrated that GBR1 and GBR2 are co-localised in the human cerebellar cortex. Together these results suggest the widespread distribution of GABA(B) receptors in human brain, and that GABA(B) receptors GBR1 and GBR2 can exist in the same cell, and therefore may function as a heterodimer in the human brain. PMID- 10814838 TI - Upregulation of CD44 expression in the retina during the rds degeneration. AB - In adult mouse retinas the standard form of the cell surface adhesion/receptor molecule CD44 is localized to Muller cell apical microvilli. In the rds (retinal degeneration slow) mouse, however, CD44 immunolabel is increased and distributed throughout the retina by 3 months postnatal. At present, it is unclear if this labeling pattern is due to the increased expression of standard CD44, the expression of variant CD44 isoforms, or an unmasking of CD44 antigenic sites. To further characterize this response, we have studied the expression of CD44 mRNA and protein in rds retinas of different ages. RT-PCR analysis demonstrated one product which represented the message for standard CD44 in adult BALB/c mouse retina and in all ages of rds retinas studied. Upon Southern blotting, this major product was detected along with two minor bands of larger size in all samples. Northern blot analysis demonstrated a major transcript of approximately 4.0 kb and a minor one of 3.0 kb in all BALB/c and rds retinas. By 3 months postnatal in rds retinas, the expression of CD44 message was increased by at least two-fold. Western blot analysis demonstrated the presence of only the standard form of CD44 protein in all BALB/c and rds retinas. An increased amount of this standard CD44 protein was observed in 2, 3, and 6 month rds retinas. Thus, this study demonstrates that the inherited retinal degeneration exhibited by the rds mouse does not cause an altered expression of retinal CD44 isoforms, but does lead to an upregulation in the expression of mRNA and protein for standard CD44. PMID- 10814839 TI - Down-regulation of orexin gene expression by severe obesity in the rats: studies in Zucker fatty and zucker diabetic fatty rats and effects of rosiglitazone. AB - Orexins (hypocretins) are lateral hypothalamic neuropeptides implicated in regulating feeding and the sleep-wake cycle. To study their possible relevance to obesity and diabetes, we measured hypothalamic prepro-orexin mRNA levels in obese, normoglycemic Zucker fatty (fa/fa) and in hyperglycemic, non-obese Zucker diabetic fatty (ZDF) rats. Hypothalamic prepro-orexin mRNA concentrations in Zucker fatty rats were 31% lower than those in lean controls (0. 69+/-0.06 vs. 1.00+/-0.10 arbitrary units, P<0.05), but did not differ between ZDF diabetic rats and non-diabetic controls. Treatment of ZDF diabetic rats with rosiglitazone (1 or 3 mg/kg body weight daily for 13 weeks) normalized plasma glucose and significantly reduced plasma insulin, while leptin levels were 67% higher than in untreated ZDF rats (20.2+/-0.5 vs. 12.1+/-2.5, P<0. 001). Rosiglitazone treatment markedly enhanced weight gain compared with untreated ZDF rats (final weight 732+/-13 g vs. 409+/-13 g, P<0. 001) even though they were restricted to the same food intake. Rosiglitazone-treated ZDF rats had significantly lower hypothalamic prepro-orexin mRNA levels (0.68+/-0.07 arbitrary units) than both non-diabetic lean controls (1.00+/-0.10, P=0.02) and untreated diabetics (1.03+/-0.14, P=0.03). Our data suggest that prepro-orexin gene expression may be suppressed by substantial weight gain. Obesity-related signals that might mediate this effect have not been identified, but plasma leptin, insulin and glucose are not obviously involved. PMID- 10814840 TI - Interleukin (IL)-10 inhibits IL-6 production in microglia by preventing activation of NF-kappaB. AB - The purpose of this study was to determine if interleukin (IL)-10 inhibits lipopolysaccharide (LPS)-induced IL-6 production in microglia by inhibiting activation of nuclear factor-kappaB (NF-kappaB). N13 microglia (a murine microglial cell line) and primary microglia from neonatal mice were cultured in the presence or absence of LPS and increasing amounts of murine IL-10 for 24 h. As predicted, LPS treatment increased supernatant IL-6 concentration in both N13 and primary microglia cultures. Pretreatment with IL-10, however, decreased LPS induced IL-6 secretion in a dose-dependent manner in both culture systems. Likewise, ribonuclease protection assays showed that LPS increased steady-state IL-6 mRNA levels, but that pretreatment with IL-10 blocked the LPS-induced increase in IL-6 mRNA. Because NF-kappaB is the predominant transcription factor responsible for IL-6 transcription in response to inflammatory stimuli, it was hypothesized that IL-10 inhibited IL-6 production by preventing nuclear translocation of NF-kappaB. Consistent with this idea, LPS increased nuclear translocation of NF-kappaB as assessed by gel mobility shift assay. Supershift assays and immunocytochemical staining showed that both the p50 and p65 subunits of NF-kappaB translocated from the cytoplasm to the nucleus upon LPS stimulation. Pretreatment with IL-10, however, inhibited LPS-induced activation of NF-kappaB. Furthermore, inhibition of NF-kappaB activity with tosyl-Phe-chloromethlyketone (a serine protease inhibitor that prevents degradation of the NF-kappaB-IkappaB complex), completely blocked LPS-induced IL-6 production. These data suggest that IL-10 inhibited IL-6 production in microglia by decreasing the activity of NF kappaB and, therefore, extend what little is known of the intricate relationship between anti-inflammatory and inflammatory cytokines in the central nervous system. PMID- 10814841 TI - Induction of 8-oxo-dGTPase activity in human lymphoid cells and normal fibroblasts by oxidative stress. AB - The pre-mutagen 8-Oxo-2'-deoxyguanosine-5'-triphosphate (8-oxo-dGTP) is formed during normal cellular metabolism and its incorporation into DNA leads to transversion mutations. Human cells possess the hMTH-1 gene encoding the enzyme 8 oxo-dGTPase, which catalyzes the hydrolysis of 8-oxo-dGTP to the corresponding 8 oxo-dGMP, preventing mutations. To elucidate the involvement of 8-oxo-dGTPase in carcinogenesis, we studied hMTH-1 gene expression and enzyme activity in response to oxidative stress to human skin fibroblasts and Jurkat cells. In fibroblasts, ranges from 0 to 100 microM H(2)O(2) caused a 2-fold induction of hMTH-1-mRNA expression and a 3-fold induction of enzyme activity. A 1.7-fold induction of mRNA expression and a 3.5-fold induction of enzyme activity was obtained in Jurkat cells after treatment ranging from 0 to 300 microM H(2)O(2). Cytotoxic concentrations of hydrogen peroxide lead to an almost complete loss of enzyme activity and an inhibition of hMTH-1 mRNA expression. Induction of hMTH-1 gene expression was prevented by addition of actinomycin D and cycloheximide. These data indicate the inducibility of the hMTH-1 gene expression and enzyme activity by prooxidative molecules, such as hydrogen peroxide. These parameters can thus be used as a marker of oxidative stress. PMID- 10814842 TI - Alterations in beta-adrenergic receptor density and adenylate cyclase activity in the rat brain treated chronically with lead. AB - Behavioral and memory impairment resulting from lead exposure is well known but the mechanism is not. We utilized the brain of lead exposed rats to investigate this problem. In an effort to elucidate the mechanism responsible for this alteration we examined blood and brain lead levels, brain beta-adrenoceptor density and cyclic AMP production in lead exposed rats. Wistar rats used in these trials were divided into six groups of ten animals each. Five groups were given drinking water containing 0.05, 0.1, 0.5, 1 and 2% lead acetate for a period of 60 days. One group (control group, 0% lead acetate) was given pure water. Application of a trend test indicated that both blood and brain lead levels increased significantly from group 0% to group 2% (group 0% 0.05). Ophthalmic examinations were normal. The result of these tests suggested normal activity of sympathetic, and hypoactivity of parasympathetic nervous system, implying parasympathetic nervous system dysfunction as a risk factor or associated finding in childhood obesity. PMID- 10814896 TI - Brain distribution and efficacy of carbamazepine in kainic acid induced seizure in rats. AB - To investigate the relationships between carbamazepine (CBZ) concentrations in serum and the brain, and the anticonvulsive efficacy in kainic acid (KA) induced seizures in rats, adult Wistar rats (n=25) were intraperitoneally given 40 mg/kg CBZ, followed by 15 mg/kg KA (n=20) or saline (control, n=5). At 90 min after the injection, CBZ concentrations in 5 rats without seizures (CBZ effective group), 5 rats with seizures (CBZ no-effective group) and five control rats were measured. Serum and brain tissues from six areas (cortex, brain stem, cerebellum, thalamus, hippocampus and striatum) were used for CBZ assay. CBZ was measured using a EMIT immunoassay kit. In blood, CBZ was higher in rats treated with CBZ+KA than in a control group (CBZ+saline). In the brain, the effective group demonstrated significantly high CBZ concentration in the hippocampus. KA appeared to raise serum CBZ level when it was given in combination with CBZ. This was probably caused by the accelerated absorption of CBZ from local site as the results of an increased metabolic rate and the more demands for blood supply after KA treatment. The positive correlation between efficacy of CBZ and the concentration in the hippocampus suggests that CBZ levels in the hippocampus is closely correlated with the efficacy of CBZ against KA induced seizures. PMID- 10814897 TI - A patient with mitochondrial myopathy associated with isolated succinate dehydrogenase deficiency. AB - We report on a boy with normal mental development who had muscle hypotonia and congenital dislocation of the hip and knee joints. Histochemical and biochemical examinations of his muscle specimen revealed no succinate dehydrogenase (SDH) activity. Since the NADH cytochrome c reductase and cytochrome c oxidase activities were normal, we concluded that he had an isolated SDH deficiency. Our patient provides further evidence for the clinical variability of this disorder. PMID- 10814898 TI - Perceptual-motor, visual and cognitive ability in very low birthweight preschool children without neonatal ultrasound abnormalities. AB - Thirty-six children born preterm with very low birth weight without neonatal brain disorders and normal cerebral ultrasound findings were examined at pre school age: visual, perceptual motor, attention, behaviour and cognitive assessments were performed in the study group as well as in a control group of term children matched for age, sex and parental educational and occupational status. The results showed a significant lower scoring in perceptual motor skills in the study group, associated with a defect of accuracy in spatial attention and a higher incidence of stereopsis impairment related with perceptual motor disabilities. Behavioural disorders, in terms of emotional maturation and hyperactivity, were significantly more frequent in the study group. To prevent behavioural and learning problems at school, a complete longitudinal assessment including visual functions and perceptual motor abilities seems mandatory in preterm born children, even in the absence of neonatal brain disorders including abnormal cerebral ultrasound findings. PMID- 10814899 TI - Benign paroxysmal torticollis of infancy. AB - Benign paroxysmal torticollis is an episodic functional disorder of unknown etiology that occurs in the early months of life in healthy individuals. The child's head tilts to one side for a few hours or days, usually without any associated symptoms. The disorder, which disappears within the first few years of life, is often misinterpreted and the patient pointlessly undergoes numerous tests. We present our series of 22 patients observed at the pediatric neurology outpatients clinic in Padova with a view to refreshing the pediatrician's memory on this frequent, benign pathology. PMID- 10814900 TI - Development of voluntary control of saccadic eye movements. I. Age-related changes in normal children. AB - To investigate the development of the voluntary control of saccadic eye movement, we examined eye movements in 99 normal children (4-13 years of age). Subjects were asked to fixate a central light for 3-5 s. A target was then presented, either to its right or left. In visually guided saccades, the mean latencies of the child group were longer than those of the adults, and decreased with age until the age of 12 where they reached adult levels. On the other hand, their peak saccadic velocities were not different from those of the adults. In the antisaccade task, they showed a higher rate of directional errors, indicating an inability to suppress reflexive saccades to the target. Mean latencies of correct antisaccades were significantly longer in the children than in the adults. Error rates and antisaccadic latency tended to decrease with age. We also examined the effects of an auditory warning signal during the fixation period and compared the results with those without. The warning stimulus was less effective in children than in adults in both tasks. Similar peak saccadic velocities between children and adults suggest the earlier development of the saccadic burst generator in the brainstem. In contrast, the delayed development of latency of saccades and antisaccades and the error rates of antisaccades suggest delayed maturation of the cerebral cortex, particularly the frontal association areas that are involved in both eye movement control and attentional processes. PMID- 10814901 TI - The serum nitric oxide levels in patients with Duchenne muscular dystrophy. AB - Nitric oxide is formed in skeletal muscle by the neuronal type nitric oxide synthase and the signalling function of dystrophin and related compounds are in part mediated by nitric oxide. Duchenne muscular dystrophy, mdx mice and patients with Becker dystrophy demonstrated neuronal type nitric oxide synthase deficiency in muscle biopsy specimens. We have intended to find out whether the plasma nitric oxide levels show any abnormality in patients with Duchenne muscular dystrophy. Serum NO levels of Duchenne patients (4.191+/-2.82 micromol/l) were significantly lower than those of the control (39.53+/-19.43 micromol/l) and cerebral palsy (77.84+/-21.70 micromol/l) groups. PMID- 10814902 TI - Bipolar affective disorder heralding cerebral demyelination in adreno-myelo leukodystrophy. AB - A young male with adrenomyeloleukodystrophy (AMLD), diagnosed at the age of 25 years, presented with signs of bipolar affective disorder (BPD) concomitant with radiological findings of central demyelination. There was a marked deterioration of the earlier relatively benign neurological dysfunction, leading to fatal bulbar syndrome. The association of BPD with central demyelination in AMLD is in agreement with previously reported cases of this and other types of multifocal central demyelination diseases. PMID- 10814903 TI - Pontocerebellar hypoplasia type 2 (PCH2): report of two siblings. AB - We describe two sisters affected by pontocerebellar hypoplasia type 2 associated with microcephaly, hypertonia, severe choreiform movements, an almost complete lack of psychomotor development, and generalized tonic-clonic seizures. Clinical and neuroradiological findings ruled out other conditions associated with pontocerebellar hypoplasia, i.e. pontocerebellar hypoplasia type 1, carbohydrate deficient glycoprotein syndrome, and olivopontocerebellar hypoplasia/atrophy. PMID- 10814904 TI - Acute transient pseudoatrophy of the brain accompanying measles infection. AB - Two infants, age 1 year, with acute encephalopathy who showed transient pseudoatrophy of the brain were reported. They each had measles infection, followed by 'mild drowsiness'. Reversible brain atrophy has been observed in patients with undernutrition, dehydration, and overdoses of steroids and valproic acid. However, these manifestations were very mild and there was no history of drug administration in our patients. The cause of the transient brain pseudoatrophy was unknown. However, activation of glial cells accompanying the measles infection was thought to be one possibility to see high value of neopterin (51 pmol/l) in the cerebrospinal fluid in one case. Although 'mild drowsiness' is not a rare manifestation in patients with measles infection, it is probable that their 'mild drowsiness' is caused by acute encephalopathy like in our cases. Thus, we recommend that computed tomography scanning or magnetic resonance imaging is performed in patients showing 'mild drowsiness' during measles infection. PMID- 10814905 TI - Ganglioside patterns in neuroepithelial tumors of childhood. AB - To elucidate a relationship between neuronal anaplasia, tumor proliferation, and ganglioside contents, we quantified gangliosides by HPTLC in tumors of neuroepithelial tissues at different grade, i. e. peripheral primitive neuroectodermal tumor (PPNET, grade IV), ependymoma (EPEN, grade III), neuroblastoma (NB, grade IV), and dysembryoplastic neuroepithelial tumor (DNT, grade I). PPNET, the most undifferentiated tumor examined had lowest concentration of total lipid-bound sialic acid. GM3 was the major ganglioside in all undifferentiated tumors (46-72.7% of total lipid-bound sialic acid). GD3 was an another component in PPNET and EPEN (7.2-17.3%). Concentration of a complex gangliosides GM1 was decreased in all tumor tissues and those of GT1a, GD1b and GT1b were decreased in tumors of high grade. The results suggest that the composition of gangliosides could be of considerable value in refining the classification of neuroepithelial tumors in infancy and childhood. PMID- 10814906 TI - A hierarchical neural network algorithm for robust and automatic windowing of MR images. AB - A novel hierarchical neural network based algorithm for automatic adjustment of display window width and center for a wide range of magnetic resonance (MR) images is presented in this paper. The algorithm consists of a feature generator utilizing both wavelet histogram and compact spatial statistical information computed from a MR image, a competitive layer based neural network for clustering MR images into different subclasses, two pairs of a radial basis function (RBF) network and a bi-modal linear estimator for each subclass, as well as a data fusion process using estimates from both estimators to compute the final display parameters. Both estimators can adapt to new kinds of MR images simply by training them with those images, which make the algorithm adaptive and extendable. The RBF based estimator performs very well for images that are similar to those in the training data set. The bi-modal linear estimator provides reasonable estimations for a wide range of images that may not be included in the training data set. The data fusion step makes the final estimation of the display parameters accurate for trained images and robust for the unknown images. The algorithm has been tested on a wide range of MR images and has shown satisfactory results. PMID- 10814907 TI - Cooperation of fuzzy segmentation operators for correction aliasing phenomenon in 3D color Doppler imaging. AB - The study described in this paper concerns natural object modeling in the context of uncertain, imprecise and inconsistent representation. We propose a fuzzy system which offers a global modeling of object properties such as color, shape, velocity, etc. This modeling makes a transition from a low level reasoning (pixel level), which implies a local precise but uncertain representation, to a high level reasoning (region level), inducing a certain assignment. So, we use fuzzy structured partitions characterizing these properties. At this level. each property will have its own global modeling. Then, these different models are merged for decision making. Our approach was tested with several applications. In particular, we show here its performance in the area of blood flow analysis from 3D color Doppler images in order to quantify and study the development of this flow. We present methods that detect and correct aliasing phenomenon, i.e. inconsistent information. At first, the flow space is partitioned into fuzzy sectors where each sector is defined by a center, an angle and a direction. In parallel, the velocity information carried by the pixels is classified into fuzzy classes. Then, by combining these two partitions, we obtain the velocity distribution into sectors. Moreover, for each found path (from the first sector to the last one), we locate and correct inconsistent velocities by applying global rules. After extracting some meaningful sector features, the fuzzy modeling, applied to the aliasing correction, makes it possible to simplify and synthesize the blood flow direction. PMID- 10814908 TI - Towards dynamic cardiac scenes interpretation based on spatial-temporal knowledge. AB - Cardiac motion analysis enables to identify pathologies related to myocardial anomalies or coronary arteries circulation deficiencies. Conventionally, bi dimensional (2D) left ventricle contour images have been extensively used, to perform quantitative measurements and qualitative evaluations of the cardiac function. Nevertheless, there are other cardiac anatomical structures, the coronary arteries, imaged on routine procedures, upon which complementary motion interpretation can be conducted. This paper presents an experimental methodology to perform dynamic cardiac scenes interpretation, studying three-dimensional (3D) coronary arteries spatial-temporal behavior. Being an alternative way to approach computer assisted cardiac motion interpretation, it reveals a wide range of rarely explored spatial-temporal situations and proposes how to address them. Considering the challenges to achieve dynamic scene interpretation, it is explained how spatial and temporal knowledge, are connected to specialist knowledge and measured parameters, to obtain a dynamic scene interpretation. Global and local motion features are modeled according to cardiac motion and geometrical knowledge, before its transformation into symbols. Anatomical knowledge and spatial-temporal knowledge are applied, along with spatial-temporal reasoning schemes, to access symbols meaning. Experimental results obtained using real data are presented. Complexity of interpretation envisioning is discussed, taking the given results as an example. PMID- 10814909 TI - The revival of surgical treatment for isolated proximal high grade LAD lesions by minimally invasive coronary artery bypass grafting. AB - BACKGROUND: Percutaneous coronary angioplasty (PTCA) and stent implantation have become the first-line intervention for patients with isolated proximal LAD lesions. Minimally invasive direct coronary artery bypass surgery (MIDCAB) has recently been developed to reduce surgical invasiveness for single LAD revascularization. This study focus on the question whether MIDCAB could be an alternative treatment for isolated proximal LAD lesions. METHODS: Starting in 1996, MIDCAB was performed in 618 patients. Angiography was performed before discharge and repeated after 6 months at follow-up examination. In an ongoing randomized trial 150 patients with an indication for treatment of a LAD lesion have been included to compare the mid-term outcome after PTCA (n=79) vs. MIDCAB (n=71). RESULTS: In 618 MIDCAB procedures 30-day mortality was 0.6%, perioperative myocardial infarction rate was 1.6%. The conversion rate to sternotomy was 3.4%. The learning curve was demonstrated by a patency rate of 96.0% in 1997, 98.0% in 1998 and 99.1% in 1999, respectively. At 6 months patency rate was 94.4% in 1997 and 97.0% in 1998. The rate of severe stenosis >75% dropped from 5.4% in 1997 to 3.4% in 1998. The over all rate of reinterventions was 5.6%. The preliminary result of the randomized trial revealed a difference in the number of perioperative adverse events, 11.4% in the MIDCAB group vs. 6.3% in the PTCA group (P<0.05). At 6 months follow-up 88. 7% of the MIDCAB patients were free from angina vs. 58.2% of the PTCA patients (P<0.02). Restenosis and a positive stress test was diagnosed in 27.9% of the PTCA patients vs. 8.4% of the MIDCAB patients (P<0.02). Reintervention was necessary in 27.9% of the patients after PTCA vs. 8.4% of the patients after MIDCAB. CONCLUSION: MIDCAB is a safe and effective but technically demanding procedure. Perioperative adverse events may be expected, but early as well as mid-term patency rate are good. When compared to PTCA, the freedom from angina and the need for additional revascularization procedures after 6 months is statistically better for patients having MIDCAB surgery. Thus, MIDCAB is considered a valuable alternative for isolated proximal high grade LAD lesions. PMID- 10814910 TI - Intraoperative flow measurement in composite Y arterial grafts. AB - OBJECTIVE: Total arterial myocardial revascularization may be achieved by using the 'Y-graft' techniques with different free arterial conduits anastomosed off the side of an in situ internal thoracic artery to reach distal coronary segments. This study was assessed to measure intraoperative graft flow, resistance and clinical outcomes. METHODS: Seventy-six patients who underwent coronary artery bypass grafting during a time period of 27 months were enrolled in this prospective study. All patients received sequential grafting by using both internal thoracic arteries, inferior epigastric and right gastroepiploic artery joined as a composite Y graft. Intraoperative graft flow, resistance and derived variables were measured. RESULTS: All patients except one showed good flow (ml/min and waveform) in either branch of composite graft. In one case, a low-flow situation through the graft was registered requiring surgical correction. Temporary occlusion of either branch did not significantly affect flow in the other side of the arterial Y. Mid-term follow-up (3 and 15 months) and angiographic studies showed a high graft patency rate. CONCLUSION: Composite arterial grafts provide excellent early and mid-term clinical results. Flow reserve of the left internal thoracic artery did not affect blood flow and resistance on either branch of the Y graft when temporary occlusion on the other side of the arterial Y was performed. PMID- 10814911 TI - Surgical angioplasty with exclusion of atheromatous plaques in case of diffuse disease of the left anterior descending artery: 2 years' follow-up. AB - OBJECTIVE: A new surgical technique of coronary artery angioplasty for diffuse and extensive lesions of the left anterior descending artery (LAD) is evaluated in this study. METHOD: Ninety-four coronary artery reconstructions (CAR) using a new technique of angioplasty of the LAD were performed: mean age of patients was 59+/-8 years, there were 21 patients with unstable angina, and 21 with recent myocardial infarction (MI). SURGICAL TECHNIQUE: Diseased LAD is bypassed with the internal thoracic artery graft (ITA). The anastomosis is made downstream from the significant proximal lesion of the LAD. A long arteriotomy (from 2 to 12 cm) is performed along the LAD up to the healthy arterial wall, followed by coverage with the onlay graft of ITA in such a fashion as to exclude the plaques from the LAD lumen. The wall of the new reconstructed LAD consisted of 75% of ITA and 25% of native LAD. The remaining part of the native LAD forms a posterior gutter giving the origins of septal and diagonal branches. RESULTS: aortic cross clamping time was 116 min, operative mortality rate was 3.2% (three patients), peri-operative infarction rate 6.6% (six patients). The follow-up was 29 months (SD=10). Of the 91 survivors, two were lost for follow-up and one died of non cardiac causes. Of the 88 patients clinically evaluated, 81 were free from angina and other cardiac events, two had new myocardial infarction in a non-grafted area, two were in congestive heart failure, and three in angina class II. Sixty patients underwent angiography. There were 57 perfect-patency CAR (95%), two ITA string sign (competitive flow), two ITA occlusions (2.5%) and no re-stenosis. CONCLUSIONS: CAR allows revascularisation of diffusely diseased LAD with acceptable operative mortality and morbidity, 2 years' good clinical results and graft patency. In this series, exclusion of plaques prevented plaque complications. PMID- 10814912 TI - Result of surgical treatments in patients with coronary-arterial obstructive disease after Kawasaki disease. AB - OBJECTIVE: To determine the efficacy of coronary artery bypass grafting (CABG) in young patients with coronary-arterial obstructive disease subsequent to Kawasaki disease. METHODS: CABG was employed in 100 patients. Age at operation ranged from 1 to 23 years at a mean of 10+/-5 years. The number of bypass grafts placed was 1 5/patient (a mean of 1.7+/-0.8). The left internal-thoracic artery (ITA) was used as a graft in 99 patients; the right internal thoracic artery in 39, the gastroepiploic artery in nine and the saphenous vein in 21. RESULTS: All patients survived the procedures. In the follow-up of 6.7+/-4.5 years, two patients died, one because of a traffic accident and the other due to sudden death. Considerable myocardial ischemia recurred postoperatively in 15, because of either obstruction of the bypass grafts or progression of other coronary-arterial obstructions. Of these, symptoms spontaneously regressed without interventional procedures in four, reoperation was indicated in four and catheter intervention was efficiently carried out in the remaining seven. Another two patients had episodes of critical ventricular arrhythmia; one of them with severe left ventricular dysfunction subsequently underwent cardiac transplantation. The patency rates of the arterial grafts were 94, 82 and 78% at 1, 5 and 10 years, respectively, and this was higher than that of the venous grafts (82, 63 and 36%, respectively). Strenuous exercise is currently prohibited in 15 patients, while the remaining 83 patients are doing well with no obvious restriction in their daily lives. CONCLUSION: Collaborating with catheter interventions, CABG using the arterial grafts can provide attractive results in patients with obstructive coronary arteries associated with Kawasaki disease. PMID- 10814913 TI - Atrial fibrillation after coronary artery bypass grafting without cardiopulmonary bypass. AB - OBJECTIVE: Atrial fibrillation is the most common complication after heart surgery. It rarely has a fatal outcome but causes patient instability, prolongs hospital stay, or even is the reason for perioperative infarction. Although conventional coronary artery bypass grafting (CABG) with cardiopulmonary bypass has excellent short-term and long-term results, the number of coronary operations on a beating heart without cardiopulmonary bypass is still growing. To reduce surgical trauma, off-pump coronary artery bypass grafting via sternotomy (OPCABG) or minimally invasive direct vision coronary artery bypass grafting (MIDCABG) via small thoracotomy are performed. The aim of this study was to estimate the frequency of atrial fibrillation in patients after myocardial revascularization without cardiopulmonary bypass. METHODS: A retrospective analysis of 48 patients undergoing myocardial revascularization without cardiopulmonary bypass was performed. Twenty-four patients underwent OPCABG and 24 were operated using the MIDCABG technique. The incidence of cardiac arrhythmias was analyzed since operation to the fourth postoperative day. Each patient had continuous ECG monitoring with option of arrhythmia analysis during ICU stay. After discharge from ICU 24-h ECG monitor studies were carried out. Surface 12-lead ECG was accomplished once a day, and additionally each time symptoms of cardiac arrhythmia occurred. Risk factors of atrial fibrillation were estimated. RESULTS: Atrial fibrillation occurred in 25% of patients after MIDCABG, in 29% after OPCABG, and in 18% after CABG with cardiopulmonary bypass. This difference has no statistical significance. Risk factors and incidence of postoperative complications were comparable in all groups. CONCLUSIONS: Atrial fibrillation is a common complication after procedures of myocardial revascularization, performed with or without cardiopulmonary bypass. The occurrence is not dependent on the type of operation. PMID- 10814914 TI - A simple way to treat chronic atrial fibrillation during mitral valve surgery: the epicardial radiofrequency approach. AB - OBJECTIVE: We describe an original radiofrequency ablation technique to treat chronic atrial fibrillation in patients undergoing mitral valve surgery. Most of the procedure is carried out epicardially, in order to avoid an undue increase of surgical time and trauma. METHODS: The ablations are performed using a temperature-controlled multipolar radiofrequency catheter. Two encircling lesions around the ostia of the right and of the left pulmonary veins are carried out epicardially, usually before cardiopulmonary bypass. Through a conventional left atriotomy the ablation procedure is completed with two endocardial lesions connecting the two encirclings between them and to the mitral valve annulus. After the mitral valve procedure is performed, the left appendage is sutured. RESULTS: From February 1998 to May 1999, 40 patients with chronic atrial fibrillation (43. 1+/-51.9 months) underwent combined radiofrequency ablation and mitral valve surgery. Mean left atrial diameter was 56.8+/-10.7 mm. Mean cardiopulmonary bypass and aortic cross-clamp time were, respectively, 119.1+/ 26.3 and 76.7+/-21.0 min. Mean postoperative blood loss was 287.2+/-186.6 ml. No reexploration for bleeding occurred. One patient died of pneumonia 12 days after operation. No patient needed permanent pacemaker implantation. Mean postoperative hospital stay was 7.3+/-5.6 days. At follow-up (mean 11.6+/-4.7 months), 30/39 (76.9%) of the patients were in stable sinus rhythm. All patients in sinus rhythm 3 months after operation recovered both left and right atrial contractility at echocardiographic control (mean 7.3+/-3.4 months). The left atrial diameter decreased significantly in patients recovering sinus rhythm. CONCLUSIONS: Epicardial radiofrequency ablation is a safe means to achieve surgical ablation of atrial fibrillation with a high success rate. The simplicity of the technique and the low procedure-related risk should dictate combined treatment virtually in all patients with atrial fibrillation undergoing open heart operations. PMID- 10814915 TI - Mitral valve surgery and atrial fibrillation: is atrial fibrillation surgery also needed? AB - OBJECTIVE: Atrial fibrillation (AF) persisting after mitral valve surgery reduces survival due to heart failure and thrombo-embolisms, and impairs quality of life. Arrhythmia surgery for AF shows today very satisfying results and therefore mitral valve surgery with AF surgery appears appealing. This study explores whether combined surgery in view of today's results of mitral valve surgery is indicated. METHODS AND RESULTS: An outcome analysis of the arrhythmia outcome of patients undergoing exclusive mitral valve surgery with or without tricuspid repair was done. Preoperative baseline characteristics including arrhythmia pattern, surgical methods and follow-up findings were reviewed. Postoperative management of AF was not protocolized. Between 1990 and 1993, 162 consecutive patients underwent mitral valve surgery; follow-up was a mean of 3.3+/-1.9 years. In-hospital and late mortality were 1 and 9%, respectively. Sinus rhythm was preserved in 40 of 57 (70%) patients with preoperative sinus rhythm whereas AF persisted in 58 of 68 (85%) of patients with preoperative chronic AF (>1 year present). Sinus rhythm without AF was observed in 10 of 29 (34%) patients with preoperative paroxysmal AF. The 4-year Kaplan-Meier survival did not differ between patients with preoperative sinus rhythm (95.2%), paroxysmal AF (89.2%) and chronic AF (82.9%) but AF persisting after surgery tended to determine survival (P=0.05). Gender, age and right ventricular pressure and tricuspid valve repair were risk factors for postoperative recurrence of AF in patients with sinus rhythm at discharge, relative risk 0.35, 1.06, 1. 04 and 2.9, respectively. CONCLUSION: Current mitral valve surgery with or without tricuspid valve repair does not eliminate preoperative paroxysmal or chronic AF. Secondly, because preoperative AF did not determine survival after mitral valve surgery, whereas postoperatively persisting AF was weakly associated with survival, atrial arrhythmia surgery primarily aims to reduce morbidity due to AF. Some characteristics can identify patients with increased propensity for persisting AF after surgery. Randomized studies of AF surgery are needed to identify suitable candidates for combined surgery. PMID- 10814916 TI - Single-stage repair of aortic coarctation with ventricular septal defect using isolated cerebral and myocardial perfusion. AB - OBJECTIVE: To avoid hypothermic circulatory arrest, we have repaired aortic coarctation with ventricular septal defect (VSD) in a one-stage procedure using an isolated cerebral and myocardial perfusion technique, and retrospectively compared this novel approach to the conventional two-stage approach. METHODS: Between October 1991 and February 1999, 24 infants, aged 4-137 days (median, 27 days) and weighing 1.7-4.3 kg (median, 3.0 kg), underwent the repair of aortic coarctation with VSD either in one (group I, n=11) or two stages (group II, n=13). In Group I, an arterial cannula for cardiopulmonary bypass was inserted into the ascending aorta in six patients with coarctation only, or into a polytetrafluoroethylene (PTFE) graft which was anastomosed to the innominate artery in the remaining five who had hypoplastic arches. A cross-clamp was placed between the innominate and left carotid arteries. The bypass flow was reduced to 30-50% of full flow at 28 degrees C, thereby maintaining a radial artery pressure of 30-45 mmHg. At this point, the aortic coarctation was repaired by an end-to end arch anastomosis, while maintaining brain perfusion and with the heart still beating. In five patients with hypoplastic aortic arches, the innominate artery proximal to the graft was then secured down and the arch anastomosis was extended to the distal ascending aorta, while providing isolated cerebral perfusion and cardioplegic arrest. After arch reconstruction was performed, the clamp was moved onto the ascending aorta, and the VSD was closed with systemic perfusion. In contrast, for group II patients, coarctation repairs were performed through a posterolateral approach, and existing VSDs were closed as secondary procedures. RESULTS: The mean isolated cerebral and myocardial perfusion time for group I was 13 min (range, 7-20 min). The myocardial ischemic time did not differ between groups I and II (43+/-4 vs. 42+/-5 min, not significant). There were no hospital mortalities or neurological complications in either group, but one late death in each group. CONCLUSION: Single-stage repair of aortic coarctation with VSD does not increase myocardial ischemic time compared to the traditional two-stage approach. The isolated cerebral and myocardial perfusion technique may offer substantial brain and myocardial protection during aortic arch reconstruction. PMID- 10814917 TI - Total cavopulmonary connection in children with body weight less than 10 kg. AB - OBJECTIVE: To evaluate the results after total cavopulmonary connection (TCPC) in small children, our clinical experience was retrospectively reviewed. METHODS: Of 164 patients undergoing TCPC, the body weight at operation was less than 10 kg (8.8+/-1.1 kg) in 54, including 21 with visceral heterotaxy. The superior caval vein (SVC) was anastomosed to the pulmonary arteries in a bidirectional fashion. To construct a channel draining the inferior caval vein (IVC), an extended polytetrafluoroethylene (ePTFE) tube was placed intraatrially (in 15 patients) or outside the heart (in 13), its diameter being 14 mm in two patients, 16 mm in 12, and 18 mm or greater in 14. A heterologous pericardial baffle was used for intraatrial rerouting in 12 patients. A pedicled autologous pericardial roll was tailored as an extracardiac conduit in 11 patients, and the pulmonary trunk was directly anastomosed to IVC orifice in three. RESULTS: Seven patients, including five with right isomerism, died in the intermediate term because of infection of the ePTFE tube in two, respiratory problems in three, atrioventricular valvar regurgitation in one, and pulmonary venous obstruction in one. Postoperative catheterization showed; SVC pressure, 11+/-2 mmHg without a pressure gradient between SVC and IVC; systemic ventricular end diastolic pressure, 5+/-2 mmHg; end diastolic volume, 122+/-54% of the anticipated normal value; ejection fraction, 0. 56+/-0.11; and cardiac index, 2.9+/-0.7 l/min per m(2). With the follow-up of 1-116 (35+/-31) months, the IVC channel has not become obstructive in all, except for one, in whom a pedicled pericardial roll was severely obstructed because of its tortuous extracardiac course crossing in front of the vertebrae. Postoperative growth was generally stable, although body weights and heights were smaller in the majority of patients when compared with the anticipated standards for Japanese children. CONCLUSION: TCPC can be justifiably established in small children. The use of autologous tissues seemed preferable for constructing the IVC channel unless anatomic orientation was unsuitable. PMID- 10814918 TI - Long-term results of sleeve lobectomy for lung cancer. AB - OBJECTIVE: Sleeve lobectomy is a lung saving procedure indicated for central tumors for which the alternative is a pneumonectomy. Current controversies relate to the safety of the procedure and adequacy as a cancer operation. The aim of the study is to analyze long-term survival after sleeve lobectomy, particularly in relation with nodal status and histological type. The incidence and patterns of recurrences were reviewed. METHODS: From 1972 to 1998, 184 patients (male 152, female 32) underwent sleeve resection for lung cancer. The mean age was 60+/-10 years (11-78 years), and the indications for operation were a central tumor (79%), peripheral tumor with nodal involvement (13%) and compromised pulmonary function (8%). The histological type was predominantly squamous (n=125, 68%), followed by non-squamous (n=50, 27%) and carcinoid tumors (n=9, 5%). Resection was complete in 161 patients (87%). RESULTS: The operative mortality was 1.6% (n=3). Follow-up was complete for the remaining 181 patients (mean, 5.7 years; range, 1 month-26 years). The survival at 5 and 10 years of all patients was 52 and 33%, respectively. Theses rates for patients with N0 status (n=97) were 63 and 48%, and 48 and 27% for those with N1 status (n=68; N0 vs. N1, P<0.05). An 8% survival rate was observed with N2 status (n=19) at 5 years, with no survivors after 7 years of follow-up. The 5 and 10 year survival was 56 and 34% for squamous carcinoma vs. 33 and 22% for non-squamous carcinoma (P<0.05). These rates were 58 and 38% for complete resection vs. 11 and 6% for incomplete resection at 5 and 10 years, respectively (P<0.05). Local recurrences occurred in 22% of cases, and the prevalence was statistically different between patients with N0 disease (14%) and N1 disease (23%; P=0.03), but not between N1 and N2 disease (42%; P=0.2). When local and distant recurrence were pooled together, the differences were highly significant between N0 (22%) and N1 (41%) disease (P=0.007), and between N0 and N2 (63%) disease (P=0.0002), but not between N1 and N2 disease (P=0.09). CONCLUSION: Sleeve lobectomy is a safe and effective therapy for patients with resectable lung cancer. The presence of N1 and N2 disease, or of non-squamous carcinoma significantly worsen the prognosis. PMID- 10814919 TI - Prognostic implications of a positive bronchial resection margin. AB - OBJECTIVE: This retrospective study evaluates probability of survival and mode of recurrence in patients with a microscopically positive bronchial resection margin following resection for primary bronchogenic carcinoma, as well as influence of radiotherapy on survival. METHODS: From January 1986 to July 1997, 40 patients had a microscopically positive bronchial resection margin following a macroscopically complete resection (17 lobectomies, three bilobectomies, four sleeve-lobectomies, and 16 pneumonectomies). Tissue diagnosis was squamous cell carcinoma in 32 patients, adenocarcinoma in four, adenosquamous carcinoma in two and neuroendocrine carcinoma in two. Lymph node status was N0 in 14 patients, N1 in 10, and N2 in 16. The bronchial margin contained carcinoma in situ in 20 patients, invasive mucosal carcinoma in five, and peribronchial infiltration in 15. All patients except the three most recent underwent adjuvant radiation therapy. RESULTS: At the conclusion of the study (January 31st, 1999), 30 patients had died: two with post-operative complications, 17 with progressive disease, ten without relation to cancer, and one under undefined circumstances. Six of 10 unrelated deaths were interpreted as respiratory complications of radiotherapy. Recurrent disease appeared in 24 patients (60%). Nineteen had progression of initial disease (47.5%): metastatic spread in 12 (30%), isolated local recurrence in four (10%), and combined local recurrence and metastases in three (7.5%). Five patients developed metachronous cancer, with bronchial location in four (10%) and laryngeal in one (2.5%). 5-year survival (Kaplan Meier) in 20 patients with carcinoma in situ was 38.7+/-13.7% (median 31 months), but rose to 55.0+/-16. 6% when excluding seven deaths not related to cancer (five of whom were secondary to radiotherapy) (chi(2)=3.080; P=0.0792). Survival in 13 patients classified N0 was 51.3+/-16.3% (median 61 months), and 71.1+/-18.0% following exclusion of unrelated deaths (chi(2)=3. 939; P=0.0472). Adverse prognosis of peribronchial infiltration was correlated to a positive N status (13 N2 and 2 N1), 5-year survival being 20.0+/-10.3% (median: 18 months). CONCLUSIONS: Prognosis of peribronchial infiltration is similar to N2 disease. In situ carcinoma does not influence survival per se. Local control of disease is probably in part due to radiotherapy. However, the high prevalence of unrelated late deaths suggests an adverse impact of radiotherapy on survival. PMID- 10814920 TI - Mucoepidermoid bronchial tumors: a review of 34 operated cases. AB - OBJECTIVE: Pulmonary mucoepidermoid tumors are commonly included with adenoid cystic carcinoma and carcinoid tumors under the misleading rubric 'bronchial adenomas'. These neoplasms are extremely rare and little is known about their oncologic behaviour. They are considered to be of high, or low malignancy. METHODS: During a 16-year-period 34 consecutive patients (24 male and 10 female with an average age of 53 years) underwent surgery for pulmonary mucoepidermoids in our clinic (0.5% of all resected lung tumors). Fourteen patients were complaint free, in the others obstructive symptoms dominated. In 23 patients the tumors were located in the upper lobes. In 24 cases lobectomy, in four instances limited resection and in six cases pneumonectomy were performed without hospital mortality. RESULTS: Twenty-nine tumors proved to be high grade and five low grade malignancy by histology. In the latest group the 5-year-survival amounted to 80% (all of these tumors were observed in stage T1-2 N0), on the other hand, however, that rate accounted only 31% at high grade malignant mucoepidermoids. There was no 5-year-survivor among patients having N2-disease (n=5). CONCLUSION: Mucoepidermoid tumors have to be treated by radical surgery with lymph node sampling and dissection. Patients with low grade tumors can be expected to be cured following complete resection, on the other hand, however, in cases of high grade malignant neoplasms surgery results in significantly worse prognosis. Careful histological typing plays a key role in prediction of late results. PMID- 10814921 TI - Evaluation of tracheo-bronchial wall invasion using transbronchial ultrasonography (TBUS). AB - BACKGROUND: Whereas accurate evaluation of tumor invasion into the tracheo bronchial wall is a critical factor in decision-making of therapy for intra thoracic malignancies, it is sometimes difficult with usual thoracic imaging techniques such as computed tomography. As recent progress in technology of ultrasonography is marked, usefulness of transbronchial ultrasonography (TBUS) in evaluation of tracheo-bronchial wall invasion was assessed. METHODS: Following routine fiberoptic bronchoscopy, an ultrasound probe (20 MHz) covered with a balloon sheath was inserted through the bronchoscope. After air present between the ultrasound probe and the tracheo-bronchial wall was eliminated with filling the balloon with distilled water, TBUS imaging was taken. RESULTS: With TBUS, normal tracheo-bronchial wall was represented as a five-layer structure at the cartilagenous portion and a three-layer structure at the membranous portion. Based on this normal TBUS imaging, tumor extent was judged in 35 patients with intra-thoracic malignancies. Among 25 patients with extra-wall tumor including esophageal cancer (n=15) and metastatic lymph nodes (n=7), tracheo-bronchial wall invasion was clearly demonstrated in nine patients, and no invasion was demonstrated in the other 16 patients. Among ten patients with tumor originating from the tracheo-bronchial wall, tumor extent beyond outer border of the wall was demonstrated with TBUS in five patients. These diagnoses were examined pathologically in 15 patients who underwent the operation, and the accuracy was 93.3%. CONCLUSIONS: It is suggested TBUS can be a useful diagnostic tool in evaluation of tumor invasion to the tracheo-bronchial wall. PMID- 10814922 TI - Pleural lavage cytology in esophageal cancer without pleural effusions: clinicopathologic analysis. AB - OBJECTIVE: The literature of pleural lavage cytology (PLC) is focused on lung cancer. We conducted this pilot study to determine the incidence of malignant pleural cytologies in patients without pleural effusions who undergo curative resection for esophageal cancer, and to evaluate the clinicopathologic significance of positive cytology. METHODS: Forty-eight patients underwent esophagectomy for thoracic esophageal cancer in our unit from January 1998 to January 1999. After thoracotomy, pleural lavage was performed before any intrathoracic manipulation and cytologically evaluated. RESULTS: There was one patient with stage I, 27 patients with stage II, and 20 patients with stage III cancer of the thoracic esophagus. The mean age was 55 years (range 41-77 years). Fifteen cases (31.3%) were found to have positive lymph nodes (N1). Squamous cell carcinoma was the dominant histopathologic type (91.7%). Positive lavage cytology in the whole group was found in 18.8% (9/48). There was no significant correlation to gender, age, clinical symptoms, histology, T or N status, TNM stage, or tumor location. CONCLUSIONS: The incidence of positive pleural lavage cytology in esophageal cancer is disconcertingly high. Positive cytology might suggest a more aggressive tumor biology. Future studies on its relation to survival and occult lymphatic metastasis are warranted. PMID- 10814923 TI - Heparin-coated circuits reduce occult myocardial damage during CPB: a randomized, single blind clinical trial. AB - OBJECTIVES: Cardiopulmonary bypass is associated with a diffuse systemic inflammatory response that can cause considerable morbidity, including organ dysfunction and bleeding. Heparin-coated circuits have been shown to give a reduced inflammatory response with clinical benefits during open-heart surgery. However, the effects on lipid peroxidation, neutrophil activation and myocardial ischemic damage in the human have remained unknown. METHODS: In a randomized single blind trial, complement activation, neutrophil counts, malondialdehyde, and cardiac enzymes were studied in 39 patients undergoing open-heart surgery. Two groups were perfused with cardiopulmonary bypass circuits with (n=20) or without heparin-coating (n=19). RESULTS: The different complement factors (C3, C4, C3d, C3d/C3 and the C-function), neutrophil levels, MDA and the cardiac enzyme levels were comparable before CPB was started and significantly increased in both groups during bypass. There were significant intergroup differences in the neutrophil levels and MDA after reperfusion (P<0.0001). Furthermore, significant positive correlations between the lipid peroxidation, expressed as MDA levels, and the levels of neutrofils and the cardiac enzyme, CK-MB were seen within the groups. CONCLUSIONS: Heparin coated circuits did lead to a decreased neutrophil response and MDA level. The correlations between CK-MB and neutrophil and MDA levels suggest neutrophil activation leading to lipid peroxidation that may influence myocardial damage. Heparin coating improved biocompatibility and was associated with less occult myocardial ischemic damage in patients undergoing open heart surgery. PMID- 10814924 TI - Fibrin gel as a three dimensional matrix in cardiovascular tissue engineering. AB - OBJECTIVE: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, cell proliferation and cell differentiation. The currently used biodegradable scaffolds in cardiovascular tissue engineering are potentially immunogenic, they show toxic degradation and inflammatory reactions. The aim of this study is to establish a new three-dimensional cell culture system within cells achieve uniform distribution and quick tissue development and with no toxic degradation or inflammatory reactions. METHODS: Human aortic tissue is harvested from the ascending aorta in the operation room and worked up to pure human myofibroblasts cultures. These human myofibroblasts cultures are suspended in fibrinogen solution and seeded into 6-well culture plates for cell development for 4 weeks and supplemented with different concentrations of aprotinin. Hydroxyproline assay and histological studies were performed to evaluate the tissue development in these fibrin gel structures. RESULTS: The light microscopy and the transmission electron microscopy studies for tissue development based on the three-dimensional fibrin gel structures showed homogenous cell growth and confluent collagen production. No toxic degradation or inflammatory reactions could be detected. Furthermore, fibrin gel myofibroblasts structures dissolved within 2 days in medium without aprotinin, but medium supplemented with higher concentration of aprotinin retained the three-dimensional structure and had a higher collagen content (P<0.005) and a better tissue development. CONCLUSIONS: A three dimensional fibrin gel structure can serve as a useful scaffold for tissue engineering with controlled degradation, excellent seeding effects and good tissue development. PMID- 10814925 TI - Analysis of the distribution of histologic myocardial lesions during acute cardiac rejection. Experimental study in rodents. AB - BACKGROUND AND METHOD: Asymmetric distribution of histologic lesions have been reported in grafted hearts that could hamper the interpretation of right ventricular endomyocardial biopsy. Heterotopic heart transplantations were performed in rats (n=59) and guinea pigs (n=20). Grafted hearts were examined by a pathologist who established the degree of cardiac rejection in the four cardiac cavities. RESULTS: Forty cardiac rejections were diagnosed in rats and ten in guinea pigs. An asymmetric distribution of histologic lesions was observed in 34 (68%) rejected hearts with greater lesions in the auricular myocardium than in the ventricular myocardium (n=25, 50%). One (n=18) or two degrees (n=7) differentiated the severity of rejection between atria and ventricles. Cardiac rejection score was significantly greater in atria than in ventricles (3.12+/ 0.18 vs. 2.6+/-0.2 (P<0.01) in rats and 2.35+/-0.37 vs. 1.6+/-0.47 (P<0.001) in guinea pigs. There were histologic lesions of rejection in the auricular myocardium in seven cases, although the ventricular myocardium was completely normal. In nine (18 %) other grafted hearts the degree of rejection was equal in the auricular myocardium and ventricular septum but was greater than the degree of rejection noted in the right and left ventricular free walls. CONCLUSION: The distribution of histologic lesions of acute cardiac rejection in rodents was heterogeneous in grafted hearts which exhibited greater lesions in the atria than in ventricles. This should be taken into consideration in the evaluation of new methods of detection of cardiac rejection and in the diagnosis of acute cardiac rejection in humans. PMID- 10814926 TI - Retrograde perfusion and true reverse brain blood flow in humans. AB - OBJECTIVE: Reversal of brain blood flow is necessary for retrograde cerebral perfusion (RCP) to have any metabolic benefit, but RCP is commonly used with little clinical evidence of the true incidence of reverse brain blood flow and impact. S-100B is exclusive to the brain and spinal cord and released during hypothermic circulatory arrest (HCA). True reverse brain blood flow (tRBBF) during RCP may be determined by demonstrating an excess of S-100B in the effluent blood from the common carotid artery compared to blood entering the brain. METHODS: Simultaneous blood samples were drawn from the jugular bulb and left common carotid artery during RCP at 5 min intervals in ten patients undergoing aortic surgery, utilizing HCA and subjected to blood gas, glucose and S-100B quantification. RCP was instituted at maximum pressure of 25 mmHg. Trans-cranial Doppler (TCD) continuously monitored the middle cerebral artery velocity (MCAV). RESULTS: The mean HCA and RCP durations were 31 min (20-50 min). Reversal of MCAV was demonstrated in only 6/10 cases (mean, 6 cm/s). Veno-arterial (V-A) extraction of oxygen and glucose occurred in all cases (P<0.001), with the mean effluent pO(2) falling to 14 mmHg. V-A S-100 gradients greater than 5% were demonstrated in 8/10 cases and correlated with higher oxygen extraction (P<0.01). In patients with and without MCAV reversal, the S-100 gradients were 1. 7 and 0.3 micromol/l, respectively (P<0.01). CONCLUSIONS: tRBBF occurred in nearly all patients. MCAV reversal appears to be a specific but insensitive guide to reverse perfusion. The de-saturation of effluent blood is not a reliable guide to true brain perfusion, and despite RCP, the brain remains ischaemic. PMID- 10814927 TI - Predicting outcome after reoperative procedures on the aortic root and ascending aorta. AB - OBJECTIVE: Reoperations on the ascending aorta after prior aortic procedures are formidable challenges. In order to identify factors predictive of clinical outcome using a uniform surgical approach, results of a 15-year experience were reviewed. METHODS: Between 1983 and 1998, 78 reoperations on the ascending aorta were performed in 71 consecutive patients. There were 56 males and 15 females, aged 54+/-13 years (10-73 years), with a mean interval to first reoperation of 60+/-76 months (5-223 months). The original operation was replacement of ascending aorta (23), aortic valve (25), aortic root (7), ascending aorta with valve preservation (9), ascending aorta and aortic valve (7). Surgical approach included femoral vessels dissection and repeat sternotomy, with femoro-femoral bypass limited to cases of traumatic reentry. Reoperation consisted in replacement of the aortic root (48), ascending aorta (15), ascending aorta and aortic valve (6), aortic root with ascending aorta and arch (6), ascending aorta and aortic arch (3). Average aortic crossclamp and cardiopulmonary bypass times were 122+/-86 and 188+/-60 min, respectively. RESULTS: Early deaths were five (7%), due to low output syndrome (3), hemorrhage (1) and sepsis (1). Mortality for emergent reoperation was significantly higher (38 vs. 3%, P=0.001). A total of 39 early complications were observed in 78 reinterventions (50%), including: traumatic reentry requiring emergent femoro-femoral bypass (4), reexploration for bleeding (4), respiratory failure (12), sepsis (5), transient neurologic dysfunction (4), renal failure (3), myocardial infarction (3), circulatory insufficiency requiring mechanical life support (2), and wound infection (2). Average intensive care unit stay was 4.5+/-9.7 days (0.5-40 days). Survival was 92+/-4%, 78+/-10% and 78+/-10% at 1, 5, and 10 years, respectively. At follow-up (mean 34+/-36 months, 1-170), survivors were in satisfactory clinical conditions (1.6+/-0. 8 mean NYHA class, 1-3) with no evidence of renal, respiratory or neurologic dysfunction. Multivariable analysis showed emergent reoperation (P=0.001), prior aortic valve replacement (P=0.005) and need for arch replacement (P=0.03) to be predictive of higher operative mortality. Longer duration of bypass (P=0.01) and aortic arch replacement (P=0.04) were predictive of higher prevalence of postoperative complications. CONCLUSIONS: Reoperations on the ascending aorta via repeat sternotomy without preventive femoral bypass are associated with low operative risk and high prevalence early complications. Emergent reintervention due to aortic dissection, particularly in patients with prior aortic valve replacement, and need for arch repair are predictive of poorer perioperative outcome. Long-term outlook of hospital survivors is satisfactory. PMID- 10814928 TI - Pumpless extracorporeal lung assist - experience with the first 20 cases. AB - OBJECTIVE: Long-term extracorporeal lung assist is limited by a significant mechanical blood trauma resulting in bleeding and hemolysis. To reduce the drawbacks of extracorporeal lung assist a new technique has been developed, by which the driving force for the extracorporeal circuit derives from the patients arterio-venous pressure gradient (pumpless extracorporeal lung assist). The aim of this clinical study was to test the feasibilty and effectiveness of pumpless extracorporeal lung assist in patients with acute respiratory distress syndrome. METHODS: Twenty patients (41+/-16 years) with acute respiratory distress syndrome of various causes and failing respirator therapy were enrolled. The minimum hemodynamic requirements included a cardiac output (CO) >6 l/min and mean arterial pressure (MAP) >70 mmHg. Pumpless extracorporeal lung assist was established using a short circuit arterio-venous shunt including a special designed low-resistance membrane oxygenator which was placed between the patients legs. RESULTS: At the time of inclusion FiO(2) in all patients was 1.0 (paO(2) 45.9+/-7 mmHg, paCO(2) 58.9+/-17 mmHg). After 24 h of pumpless extracorporeal lung assist FiO(2) was reduced to 0.8+/-0.1. A significant improvement in oxygenation (paO(2) 84.1+/-21 mmHg, P<0.05) and CO(2) removal (paCO(2) 32.7+/-5 mmHg, P<0.05) was notable. The mean extracorporeal flow was 2.6+/-0.6 l/min, which represented approximately 25% of the patients mean CO (10.8+/-2 l/min). The median assist time was 12+/-8 (1-32) days. Fifteen out of twenty patients were weaned off pumpless extracorporeal lung assist. Five out of twenty patients died while on the system (four sepsis, one ventricular fibrillation). Three out of twenty patients died after successful weaning on day 8, 30, and 50, respectively. Twelve out of twenty patients were discharged in a healthy state (overall survival 60%). Technical problems included thrombosis of the venous cannula (n=5), thrombus formation within the membrane oxygenator (n=2), membrane oxygenator plasma leakage (n=2), and membrane oxygenator contamination with Candida albicans. No bleeding complication was observed. CONCLUSION: Pumpless extracorporeal lung assist is feasible and effective in a selected group of patients with acute respiratory distress syndrome but preserved hemodynamic function. By eliminating the pump and reducing the tubing length blood trauma can be minimized. Being very simple the system entails fewer risks of technical complications and also facilitates nursing care. PMID- 10814929 TI - Dissecting thoracoabdominal aortic aneurysm associated with an isolated right sided aortic arch. AB - Although a right-sided aortic arch is not a rare anomaly, an aortic dissection involving an isolated right-sided aortic arch is extremely rare and remains a complicated entity for surgical therapy because of its anatomical characteristics. Previous reports that we have identified in English literature include only six surgical cases of aortic dissection involving a right-sided aortic arch. We report on a 75-year-old female who had a chronic thoracoabdominal aortic aneurysm following type B aortic dissection in a right-sided aortic arch. Graft replacement including reconstruction of Adamkiewicz artery and a celiac trunk was performed. The postoperative course was uneventful except for a prolonged ventilatory support. This case seems to be the first successful case of extended graft replacement for this pathology. PMID- 10814930 TI - Aorto-atrial fistula after operated type A dissection. AB - The development of a fistula between the aorta and right atrium is a rare complication of ascending aortic dissection and has a high mortality if not diagnosed and surgically treated. Clinical diagnosis is best supported by specialised imaging. In addition it may present technically very challenging problems. We report the first case which follows aortic root replacement for an acute type A dissection. Aorto-right atrial fistula (AoRAF) rarely complicates ascending aortic dissection. We report the first case to follow corrective surgery for aortic dissection. PMID- 10814931 TI - Transmyocardial jet after percutaneous transmyocardial revascularization. PMID- 10814932 TI - TTV - a virus searching for a disease. AB - TTV is a new virus which was identified in the serum of a patient with non-A-G post-transfusion hepatitis in Japan. The original workers aimed to account for the small number of post transfusion hepatitis cases found in their clinical practice. Subsequent work has attempted to determine the properties and unravel the natural history of the new agent. The original study applied representational difference analysis to detect foreign DNA sequences which were present only during the acute phase of illness. Subsequent studies have used PCR to study the agent in serum liver and faeces. This review summarises the published data from clinical and epidemiological studies in different countries. The inclusion of the virus in the parvovirus family seems premature because its size is unknown, its reported density is too light and its sequence lacks the characteristic long terminal repeats. The agent can be found in 1-40% of health blood donors in different countries and also in faeces. TTV is ubiquitous but its taxonomic identity and disease load remain to be determined. PMID- 10814933 TI - Viral load trend in HIV-1 seropositive patients with different CD4 cell counts before starting HAART. AB - BACKGROUND: The efficacy of highly active antiretroviral therapy (HAART) was prospectively monitored in HIV-1 seropositive patients by analyzing HIV-1 RNA viral load. OBJECTIVES: The aim of the study was to evaluate the viral load course in two different groups of patients (group 1: CD4>400/mm(3), group 2: CD4<200/mm(3)) during a period ranging from 9 to 25 months. STUDY DESIGN: HIV-1 viral load, at the start and during HAART, was analyzed in 117 patients who had previously been treated only with two anti-transcriptase drugs but were naive for protease inhibitors. RESULTS: The results showed that, after the beginning of therapy, high plasma HIV-1 RNA levels dropped to undetectable values (<50 copies HIV-RNA/ml) in one third of patients over a mean period of about 9 months irrespective of the initial CD4 cell count, even though a viral reduction of at least 2log(s) in a significantly shorter period of time (P<0.001) was observed only among patients who began retroviral therapy with a higher CD4 cell count. CONCLUSION: The response to HAART was not dramatically affected by the initial CD4 count. Though restricted to a small number of subjects, the data support the idea that therapeutic intervention can be effective even in an advanced stage of HIV-1 infection, when patients show a decreased number of CD4 T-lymphocytes. PMID- 10814934 TI - An outbreak of enterovirus 71 infection in Taiwan 1998: a comprehensive pathological, virological, and molecular study on a case of fulminant encephalitis. AB - BACKGROUND: In a recent enterovirus outbreak in Taiwan, serotype 71 was the culprit of encephalitis causing rapid clinical deterioration and death among young children. OBJECTIVES: Since knowledge of enterovirus 71 (EV71) infection in the central nervous system is still limited, the purpose of the present case study was attempted to uncover the pathogenesis of the virus. STUDY DESIGN: We performed a detailed pathological examination, virological and molecular studies on a case of EV71 infection with a rapidly fatal outcome. In addition, the whole genome of the virus was sequenced to determine the genetic relationships to other enteroviruses and two other EV71 strains (a prototype BrCr and a neurovirulent MS strain), and to provide the genetic basis of its neurovirulence of the new isolate, NCKU9822 strain. RESULTS: Characteristic features of acute encephalomyelitis were observed, with most prominent lesions in the spinal cord and brain stem. Mild myocarditis and pancreatitis were also noticed. EV71 antigen was localized to neurons on immunohistochemical staining. EV71 was recovered from all organs with inflammatory reaction. Sequence analysis showed that overall NCKU9822 and the two EV71 strains shared 80% nucleotide identity and 95% amino acid identity. It had only 45% amino acid and 52% nucleotide identities with polioviral P1 capsid region. CONCLUSION: The spinal cord and brain stem were the main targets of EV71 in the fatal cases in this outbreak, however, heart and pancreas might also be involved. Since the amino acid sequences in the P1 region are conserved (97% identity) among the three EV71 strains as compared to other enteroviruses and polioviruses, these EV71 neurovirulent strains might share the same mechanisms of neurovirulence, and the mechanisms might be different from those in polioviruses. PMID- 10814935 TI - An outbreak of enterovirus 71 infection in Taiwan, 1998: epidemiologic and clinical manifestations. AB - BACKGROUND: An outbreak of enterovirus infections occurred throughout Taiwan in 1998. The diseases were manifectated with hand, foot, and mouth disease (HFMD), some associated with meningitis, encephalitis, or acute flaccid paralysis (AFP). OBJECTIVES: This study is aimed to characterize and analyze the epidermologic and clinical features during the outbreak. STUDY DESIGN: The epidemiologic information was collected from the Ministry of Health on passive surveillance; clinical and virological investigations were carried out at National Cheng Kung University Medical Center. RESULTS: Between April and December 1998, 405 children were hospitalized, and 78 patients died during this outbreak in Taiwan. There were 119 cases identified to be EV71 infection in Tainan and Chiayi areas; 105 cases by virus isolation and 14 by serological assay. The outbreak had a biphasic curve with peak in June and October, especially in the southern Taiwan. Seventy two percent of patients were below 3 years of age. The spectrum of disease included HFMD in 54, HFMD with central nerve system (CNS) involvement in 37, herpangina in 12, aseptic meningitis in three, encephalitis/ meningoencephalitis in ten, acute flaccid paralysis in three. There was nine fatal cases complicated with neurogenic pulmonary edema. Myoclonus with sleep disturbance was the most important early sign of EV71 infection with CNS involvement. CONCLUSION: Our experience demonstrated that the EV71 isolated in Taiwan had strong dermatotropic as well as neurotropic tendencies. Early detecting CNS involvement and commencing aggressive therapy may reduce the mortality. PMID- 10814936 TI - Virological diagnosis of herpes simplex encephalitis. AB - BACKGROUND: The herpes simplex encephalitis (HSE) represents one of the most severe infectious diseases of the central nervous system. As effective antiviral drugs are available, rapid and reliable diagnosis has become important. OBJECTIVES: To evaluate retrospectively the usefulness of polymerase chain reaction (PCR) as well as serological procedures for the diagnosis of HSE. STUDY DESIGN: 631 cerebrospinal fluids (CSF) from patients with clinical suspicion of encephalitis were tested for type-specific herpes simplex virus (HSV) DNA using PCR. Virus-specific antibodies including their intrathecal synthesis were measured in 624 CSF and 2409 serum samples of 2711 patients suspected of having encephalitis. RESULTS: Positive results were obtained by PCR in eight patients (1. 3%) for HSV-1 and in seven (1.1%) for HSV-2. Intrathecal antibody synthesis was estimated in 24 (3.8%) patients. In general, no intrathecal antibodies could be measured in patients with positive PCR results and vice versa the intrathecal immune response became positive when CSF was cleared from the HSV. Results of the antibody detection in serum specimens revealed an active HSV infection in 268 out of 2367 patients (11.3%). CONCLUSIONS: The detection of HSV-DNA by PCR is the method of choice for diagnosis of HSE in the early phase of the disease. During the later stage, it has to be diagnosed by the estimation of intrathecally synthesized antibodies. PMID- 10814937 TI - Presence of herpes simplex virus (HSV) in peripheral leukocytes of patient who developed active HSV infection after bone marrow transplantation. AB - BACKGROUND: Despite of prophylactic antiviral therapy, latent HSV may be reactivated in bone marrow transplant (BMT) recipients and cause serious disease. Rapid diagnosis of HSV infection is needed to prompt institution of appropriate therapy. OBJECTIVES: We report a case of the allogenic BMT recipient, who developed ulcerative esophagitis which progressed to generalized HSV infection and graft versus host reaction (GVHR).We consider several diagnostic approaches to detection of active HSV infection in this patient. STUDY DESIGN: Polymerase chain reaction (PCR) was used to detect HSV DNA in esophageal biopsy specimens and peripheral leukocytes (PBL). Isolation of HSV in tissue culture was performed to prove infectious virus in swabs from mucocutaneous lesions or in PBL. RESULTS: Using PCR, HSV DNA was detected in peripheral leukocytes of the patient who had developed generalized HSV infection accompanied with hepatosplenomegaly and hepatitis. At that time, a fully infectious ACV-resistant HSV was isolated from his PBL. On the other hand, HSV DNA was not detected in PBL of other BMT recipients with skin- or organ-localized infection. CONCLUSIONS: Presence of HSV DNA in PBL of BMT recipients can signalize generalized HSV infection. Isolation of HSV from PBL by cocultivation with human fibroblasts can be used as an alternative diagnostic approach in these patients. PMID- 10814938 TI - TT virus: a study of molecular epidemiology and transmission of genotypes 1, 2 and 3. AB - BACKGROUND: TT virus (TTV) is a recently discovered virus, which is not related to any other known virus infecting humans. OBJECTIVES: To investigate: (i) the world-wide distribution of the three major TTV genotypes; and (ii) the possible routes of viral transmission. STUDY DESIGN: (i) The phylogenetic distribution of 494 TTV isolates originating from 31 countries was analysed, using partial ORF1 sequences. (ii) Faeces samples (n=22) and saliva samples (n=72) from French individuals were tested for the presence of TTV DNA. (iii) Viral titres in paired serum and saliva samples were compared. RESULTS: (i) Genotypes 1, 2 and 3 were distributed world-wide, with a high proportion of type 1 in Asia (71%) and no type 3 identified in Africa to date. In the USA, 77% of isolates were grouped in four clusters only (genetic distances <10%). This was also the case of 76% of French isolates, 76% of Japanese isolates, and 89% of Hong Kong isolates. (ii) TTV DNA was detected in 18% of faeces samples and 68% of saliva samples tested. (iii) Viral titre in saliva samples was 100-1000 times higher than that of the corresponding serum. CONCLUSIONS: (i) The observed epidemiological distribution of TTV isolates is compatible with an ancient dissemination of viral ancestors belonging to the different genotypes and a slow genetic evolution in sedentary populations. (ii) Besides the possible transmission of TTV by the parental and oral-faecal routes, the high titre of TTV DNA observed in saliva raises the hypothesis of the viral transmission by saliva droplets. This route of transmission could explain the high degree of exposure to viral infection observed in the general population. PMID- 10814939 TI - Cohort effect of HCV infection in liver cirrhosis assessed by a 25 year study. AB - BACKGROUND: The role of HCV infection in the development of chronic liver disease is still unclear. OBJECTIVES: Assess the presence of HCV infection in patients with liver cirrhosis. STUDY DESIGN: 123 cases of cirrhotic liver randomly selected over a 25 years period (1969-1994) from the autopsy archives of the Pathology Department of the University of Trieste, Italy, were analyzed for the presence of HCV viral genome. METHODS: Total RNA was extracted from formalin fixed paraffin-embedded tissues of the cirrhotic liver. Genotype analysis for HCV was performed after RT-PCR by dot-blot hybridization with the three major genotype-specific probes (G1, G2 and G3). RESULTS: The overall HCV genome frequency was 50.4% (62/123). The positivity was quite constant in the 1969-1979 period (35-38%), rose to 65% in 1984, peaked to 77% in 1989 (P<0.005 vs. the previous decade), and decreased to 50% in 1994. HCV genotype G1 was found in 89% of the 62 positive samples. The mean age of death of HCV-positive and HCV negative patients was comparable (69+/-12 vs. 67+/-16 years, NS). CONCLUSIONS: These data show an increasing frequency of HCV infection in cirrhotic liver tissues from 1969 to 1994, which peaked in 1989. The genotype G1 was the almost uniquely associated with cirrhosis. These findings indicate that the HCV infection occurred around the late 1950s-early 1960s, thus supporting the hypothesis of a cohort effect. HCV infection seems not to alter the natural history of liver cirrhosis as indicated by the comparable age at death of HCV positive and HCV negative patients. PMID- 10814940 TI - Evaluation of a prototype Amplicor PCR assay for detection of human immunodeficiency virus type 1 DNA in blood samples from Tanzanian adults infected with HIV-1 subtypes A, C and D. AB - BACKGROUND: In previous evaluations, the standard Amplicor HIV-1 DNA PCR test (Roche Diagnostic Systems) has been reported to have low sensitivity for the detection of some non-B HIV-1 subtypes. It has therefore become necessary to determine the performance of commercially available as well as prototype HIV-1 PCR assays for HIV-1 DNA detection in samples from various geographical settings, in order to assess their ability to detect the different HIV-1 genotypes. OBJECTIVES: To determine the performance of the prototype Roche Amplicor version 1.5 PCR test in comparison to that of the standard Roche Amplicor PCR test for the detection of HIV-1 DNA in blood samples from HIV-1 seropositive pregnant Tanzanian women infected with various HIV-1 subtypes. STUDY DESIGN: This was a cross-sectional study done on 161 blood samples collected from 106 HIV-1 seropositive and 55 seronegative asymptomatic pregnant women attending antenatal clinic in Dar es Salaam, Tanzania. METHODS: Cell pellets for PCR were prepared from EDTA blood by the Amplicor whole blood PCR sample preparation method. Plasma was used for HIV serology by enzyme linked immunosorbent assays. Subtyping was done by the heteroduplex mobility assay (HMA) using cell pellets and/or plasma. RESULTS: The sensitivities of the prototype PCR and the standard assays were 99.1% (105/106) and 97% (99/102), respectively. All samples from 55 HIV-1 seronegative women were negative by both PCR assays. Among the 101 samples subtyped by HMA, 48 (47%) were subtype A, 30 (30%) subtype C, 20 (20%) subtype D and 3 (3%) were indeterminate. In the standard DNA PCR assay, a statistically significantly higher proportion of subtype A samples had a low level of reactivity as measured as optical density compared with the subtypes C and D samples while in the prototype assay all three subtypes showed a high level of reactivity. CONCLUSIONS: The Amplicor version 1.5 DNA PCR test has a high sensitivity for the detection of HIV-1 DNA in blood samples from Tanzanian adults. Since performance of this assay does not appear to be influenced by differences in HIV-1 subtypes A, C and D, it has the potential for use in the detection of HIV-1 DNA in samples from geographic areas where these subtypes are prevalent. PMID- 10814942 TI - Exploring the wound research abstracts: WOCN conference 1999 PMID- 10814941 TI - First experiences with an accelerated CMV antigenemia test: CMV Brite Turbo assay. AB - BACKGROUND: Cytomegalovirus disease is still a major problem in immunocompromised patients, such as bone marrow or kidney transplantation patients. The detection of viral antigen in leukocytes (antigenemia) has proven to be a clinically relevant marker of CMV activity and has found widespread application. Because most existing assays are rather time-consuming and laborious, an accelerated version (Brite Turbo) of an existing method (Brite) has been developed. The major modification is in the direct lysis of erythrocytes instead of separation by sedimentation. OBJECTIVE: In this study the Brite Turbo method has been compared with the conventional Brite method to detect CMV antigen pp65 in peripheral blood leukocytes of 107 consecutive immunocompromised patients. RESULTS: Both tests produced similar results. Discrepancies were limited to the lowest positive range and sensitivity and specificity were comparable for both tests. CONCLUSIONS: Two major advantages of the Brite Turbo method could be observed in comparison to the original method: assay-time was reduced by more than 50% and only 2 ml of blood was required. An additional advantage was the higher number of positive nuclei in the Brite Turbo method attributable to the increased number of granulocytes in the assay. Early detection of CMV infection or reactivation has become faster and easier with this modified assay. PMID- 10814944 TI - The nurse's role in prostate cancer treatment: challenge or opportunity? PMID- 10814943 TI - President's message: finding the opportunities amidst the crises (or, get out the lemonade machine!). PMID- 10814945 TI - Postprostatectomy incontinence: the magnitude of the problem. AB - Prostate cancer is being detected earlier in its development as a result of heightened public awareness and better screening and assessment techniques. Localized prostate cancer is often surgically treated, leaving many men incontinent of urine on either a short-term or long-term basis. This article explores the epidemiology of prostate cancer and the prevalence and incidence of urinary incontinence in men after they have undergone radical prostatectomy. Implications for WOC nursing research and practice are addressed. PMID- 10814946 TI - Managing urinary incontinence following radical prostatectomy. AB - New diagnoses of prostate cancer more than tripled between 1990 and 1996, largely because of improved methods of detection and heightened public awareness. Radical prostatectomy is often undertaken in men with prostate cancer who are expected to live at least 10 more years and have tumors confined to the prostate gland. Because of high 10-year survival rates, the demand for radical prostatectomy has increased steadily during the past decade and continues to climb as men pursue a cure and attempt to maximize survival. Survival benefits aside, however, radical prostatectomy carries a significant risk of urinary incontinence, which can dramatically impair quality of life. Despite medicine's steadfast pursuit of continence-preserving approaches to radical prostatectomy, nurses are currently challenged to provide care for a growing cohort of patients with postprostatectomy incontinence. This article reviews the clinical presentation and pathophysiology of postprostatectomy incontinence, including assessment and treatment options. PMID- 10814947 TI - Urinary incontinence as a factor in prostate cancer treatment selection. AB - Urinary incontinence is a common adverse effect associated with treatment for early stage prostate cancer. The influence of this factor on treatment selection decisions by patients and their partners has been explored only minimally in the literature. Data regarding the actual incidence of incontinence associated with prostate cancer treatment are confusing because of the lack of standardized definitions of incontinence. Radical prostatectomy is associated with higher rates of urinary adverse effects than is radiation therapy. Brachytherapy appears to be associated with a low risk of incontinence, whereas cryosurgery is associated with significant urinary adverse effects. Including incontinence, urethral sloughing, and bladder neck obstruction. The influence of these adverse effects on decision making regarding prostate cancer treatment selection is difficult to ascertain. Research indicates that both men and their partners appear to have difficulty processing information presented to them regarding the probability of urinary adverse effects and the degree to which these adverse effects may have an impact on their daily lives. PMID- 10814948 TI - Voiding dysfunction after radiation to the prostate for prostate cancer. AB - Radiation therapy is an increasingly common treatment for prostate cancer. Although radiotherapy is generally effective, it is not free of complications. Acute adverse effects of radiation are usually mild and managed on an outpatient basis. In contrast, long-term complications, although uncommon, may be debilitating or even life-threatening, requiring more invasive treatments or hospitalization. This article will review the pathophysiology of the acute and long-term effects of radiation on the lower urinary tract and implications for WOC nursing management of patients with voiding dysfunction following radiation therapy for prostate cancer. PMID- 10814949 TI - Stigma associated with postprostatectomy urinary incontinence. AB - PURPOSE: This collective case study explored the social implications of postprostatectomy urinary incontinence (UI). DESIGN: Collective case study. SETTINGS AND SUBJECTS: Three men older than age 60 years dwelling in the community who had urinary incontinence following prostatectomy. METHODS: Unstructured, in-depth interviews were thematically analyzed and presented in the form of a collective case study. RESULTS: Participants articulated two separate entities: a private and public identity. In their public identity, the participants went to great lengths to appear as a person who was continent of urine, and they expressed fear that their UI would be exposed. In revising their private identity, men used knowledge of their anatomy and physiology, family history, and life events to reject the cultural attitudes towards UI and renegotiate a new sense of self that was accepting of their leaking body. CONCLUSION: A stigma exists for men who experience UI following prostatectomy; this stigma affects public and private identity. Naturalistic inquiry methods such as the collective case study described here provide new knowledge for continence nurses as they assist these patients to manage their UI within a social context. PMID- 10814950 TI - Quality of life in men with urinary incontinence after prostate cancer surgery. AB - Quality of life assessment is significant to health care providers because it helps us understand the experience of well-being as it relates to an illness and its severity, symptoms, and co-morbidities. Attempting to deduce the influence of illness on quality of one's life is complex; however, this area of research has demonstrated that the measurement of quality of life is as important in providing comprehensive care as the treatment itself. Prostate cancer is the most prevalent cancer in American men. Radical prostatectomy is frequently considered the treatment of choice for localized prostate cancer. Despite its widespread use, considerable morbidity exists, including erectile dysfunction and urinary incontinence. Although not all men who undergo radical prostatectomy will experience urinary incontinence, those who do find that it influences their daily lives, affecting the clothes they wear, their activities, sleep patterns, social relationships, and self-esteem. Based on the compelling nature of this problem, this article will focus on the effects that urinary incontinence has on the quality of life in men who undergo surgical treatment for prostate cancer. PMID- 10814951 TI - A review of radiation proctitis in the treatment of prostate cancer. AB - Pelvic radiation is a common therapy for the treatment of prostate cancer. A complication of this therapy, radiation proctitis, may be limited to the direct posttreatment period or it may appear as serious complications that occur months to years after therapy has been completed. Mucosal damage, present with both acute and chronic radiation proctitis, produces a variety of symptoms including mucoid diarrhea, pain upon defecation, serious rectal bleeding, stenosis, and fistula formation. The treatment of radiation proctitis is symptom related, and the goals of therapy include the prevention or correction of mucosal changes and eradication of rectal bleeding. This article will review the pathophysiology of radiation proctitis and its treatment. PMID- 10814952 TI - After effects: tackling the morbidity of prostate cancer treatment. PMID- 10814953 TI - Comparative, observational designs: case-control and cohort studies. PMID- 10814954 TI - Postprostatectomy urinary incontinence managed by behavioral methods. PMID- 10814955 TI - Poly(ADP-ribose) polymerase in DNA damage-response pathway: implications for radiation oncology. AB - Poly(ADP-ribose) polymerase (PARP) catalyzes the transfer of successive units of ADP-ribose moiety from NAD(+) covalently to itself and other nuclear acceptor proteins. PARP is a zinc finger-containing protein, allowing the enzyme to bind to either double- or single-strand DNA breaks without any apparent sequence preference. The catalytic activity of PARP is strictly dependent on the presence of strand breaks in DNA and is modulated by the level of automodification. Data from many studies show that PARP is involved in numerous biological functions, all of which are associated with the breaking and rejoining of DNA strands, and plays a pivotal role in DNA damage repair. Recent advances in apoptosis research identified PARP as one of the intracellular "death substrates" and demonstrated the involvement of polymerase in the execution of programmed cell death. This review summarizes the biological effects of PARP function that may have a potential for targeted sensitization of tumor cells to genotoxic agents and radiotherapy. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 59-67 (2000). PMID- 10814956 TI - Atomic force microscopy examination of conformations of polynucleotides in response to platinum isomers: significance of GC content at broken ends. AB - Atomic force microscopy is a technique that enables visualization of macromolecular conformations of polynucleotides at nanometer resolution. We investigated the results of interactions of cisplatin, a DNA binding anticancer drug, and its inactive counterpart, transplatin isomer, on the molecular conformation of polynucleotides: poly d(G-C). poly d(G-C) (polyGC) and poly d(A T). poly d(A-T) (polyAT). We observed that polyAT exhibited an increased number of enlarged ends of molecules, which we attribute to unwound and/or collapsed regions of polyAT. PolyGC molecules did not show such ends unless cisplatin was added to the PolyGC polymers. Transplatin had the apparent effect of causing overlapping or stacking of the polymer molecules. Addition of exonuclease-III to these polymers removed the visible enlarged ends. The effects of cisplatin as compared to transplatin on the polyGC duplex polymers provide support for the presence of intrastrand covalent linkages, consistent with known N7 guanine interaction of the cis isomer on molecular conformation. Furthermore, our results indicate that the mechanism of interactions of DNA with cisplatin may be dependent on the GC content of the molecules. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 68-72, 2000. PMID- 10814957 TI - Dose response of amifostine in protection of growth plate function from irradiation effects. AB - The dose-response radioprotectant effects of amifostine on rat growth plate have not been studied. The purpose is to examine the relative effects of varying doses of amifostine on functional damage to the Sprague-Dawley rat growth plate from a single fraction radiation exposure. Thirty-six weanling Sprague-Dawley rats underwent single dose 17.5 Gy radiation exposure to the right knee. The contralateral left limb served as the nonirradiated control. Six groups of six animals each received, 20 minutes before radiation exposure, intraperitoneally administered amifostine at the following doses: 0, 50, 100, 150, 200, and 250 mg/kg. Six weeks after treatment, the rats were euthanized and the lower limbs disarticulated, skeletonized, radiographed, and measured. Statistically significant dose-related differences were observed between amifostine dosage groups for mean right-side growth, growth-loss, and limb-length discrepancy. The mean right-side growth recovered by amifostine administration increased from 14% at 50 mg/kg to 57% at 250 mg/kg. Growth-loss and limb discrepency were significantly reduced in proportion to increasing amifostine doses. Despite these positive effects of amifostine, amifostine associated mortality was identifiable beginning at 200 mg/kg and increased rapidly thereafter. This report suggests a directly proportional relationship between amifostine dose and its protective effects on the growth plate. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 73-79 (2000). PMID- 10814958 TI - Improved results with accelerated hyperfractionated radiotherapy of advanced head and neck cancer. AB - A retrospective study on local tumor control, survival, and complications of conventional irradiation vs. accelerated hyperfractionated irradiation in patients with selected head and neck cancer sites was undertaken. A total of 1,007 consecutive patients treated with radiation alone for cure from 1974-1997 were analyzed. Excluded were female patients, patients with T1 stage of the vocal cord, and patients also treated with brachytherapy implants. There were 637 patients treated with conventional fractionation once daily (QD) in 1974-1997, at a median 2.1 Gy/fraction, to a total median dose of 71.4 Gy in a median overall time of 54 days. As was common before the mid-1980s, 39% and 22% of patients had overall times exceeding 8 and 9 weeks, respectively; 370 patients were treated with accelerated hyperfractionation twice daily (BID) from 1987-1997, at a median of 1.6 Gy/fraction, with an interfraction interval of 4-6 h, to a total median dose of 68 Gy in 40 days. Both schedules were well-balanced with respect to their pretreatment characteristics. Patients were not randomized into QD or BID. The 10 year actuarial probability of local control was 37% vs. 56% for QD and BID, respectively (P < 0.001), which reflects an increase of 19% or a 51% reduction in the local failure rate. Multivariate analysis revealed that T-stage, QD or BID schedule, and overall treatment time were significant independent factors for achieving local tumor control. The 10-year actuarial probability of cause specific disease-free survival was 25% and 30% for QD and BID, respectively (P = 0.012). Acute morbidity was slightly higher with the BID schedule: patients requiring tube or parenteral feeding were 2.4% for BID and 0.5% for QD (P = 0.01). The 10-year actuarial probability of RTOG/EORTC Grades 3-5 late effects was 13% for both QD and BID. The lack of increase in late complications was most probably due to the lower total dose and dose per fraction in the BID schedule. This study has shown that accelerated hyperfractionated irradiation using two doses of 1.6 Gy each treatment day for less than 6 weeks in advanced head and neck cancer in male patients provides significantly better local tumor control and cause-specific disease-free survival, without increased late morbidity, than conventional fractionation delivered at the previously relaxed overall times of 7 weeks, but sometimes exceeding 8 or 9 weeks. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 80-91, 2000. PMID- 10814959 TI - Intensity-modulated radiation therapy in head and neck cancers: The Mallinckrodt experience. AB - The purpose of the study was to investigate the feasibility and the optimization of tomotherapy-based intensity-modulated radiation therapy (IMRT) in patients with head and neck cancer. From February 1997 to November 1997, 17 patients with squamous cell carcinoma of the head and neck were treated with IMRT. Patients were immobilized with a noninvasive mask and treated using a serial tomotherapy device on a 6 MV linear accelerator. Treatment planning was performed on a Peacock inverse planning system and prescription optimization was used to achieve the best plan for target coverage and parotid sparing. The treatment planning system process has a dosimetric characteristic of delivering different doses to different target structures simultaneously in each daily treatment; therefore, the biological equivalent dose was implemented using the linear-quadratic model to adjust the total dose to the target volume receiving a daily dose of less than 1.9 Gy. All eight patients with gross disease (six in the nasopharynx, two in the tonsil) and one patient with recurrent nasopharyngeal carcinoma received concurrent cisplatin chemotherapy. Six postoperative patients were treated with irradiation alone. Median follow-up was 2.2 years (range 2.6-1.8 years). All patients completed the prescribed treatment without unexpected interruption. Acute side effects were comparable to those of patients treated with conventional beam arrangements during the same period. No patient required gastrostomy during irradiation. The preliminary experience showed that the noninvasive immobilization mask yielded high positioning reproducibility for our patients. To spare the parotid gland, which is in the proximity of the target, a fraction of the target volume may not receive the prescribed dose. In the best-achievable plan of our studied cohort, only 27% +/- 8% of parotid gland volumes were treated to more than 30 Gy, while an average of 3.3% +/- 0.6% of the target volume received less than 95% of the prescribed dose. This is mainly related to the steep dose gradient in the region where the target abuts the parotid gland. The inverse planning system allowed us the freedom of weighting normal tissue-sparing and target coverage to select the best-achievable plan. Local control was achieved in eight patients with gross tumor; six were treated postoperatively. Of three reirradiated patients, two had symptomatic improvement but persistent disease, and one is without evidence of disease. In summary, a system for patient immobilization, setup verification, and dose optimization for head and neck cancer with parotid sparing without significantly compromising target coverage is being implemented for a tomotherapy-based IMRT plan at the Mallinckrodt Institute of Radiology. The initial clinical experience in tumor control is promising, and no severe adverse acute side effects have been observed. Further refining of delivery technology and the inverse planning system, gaining clinical experience to address target definition and dose inhomogeneity within the targets, and understanding the partial volume effect on normal tissue tolerance are needed for IMRT to excel in the treatment of head and neck cancer. Int. J. Cancer (Radiat. Oncol. Invest.) 90, 92-103, (2000). PMID- 10814960 TI - Use of radiation therapy in posttransplant lymphoproliferative disorder (PTLD) after liver transplantation. AB - Posttransplant lymphoproliferative disorder (PTLD) is a common and life threatening complication of immunosuppression used to prevent rejection of solid organ and bone marrow transplants. There is no standardized treatment algorithm, but numerous management strategies are available. We describe a patient who developed a solitary lymphoproliferative lesion in the porta hepatis 9 months after orthotopic liver transplant. Following reduction in immunosuppression with no response, she was treated with involved field radiotherapy utilizing CT-based treatment planning. A partial radiographic response was obtained, and she has not developed disease in the engrafted liver or systemically. Based on the present case report, involved field radiotherapy seems to be a reasonable treatment option for patients with localized PTLD. Int. J. Cancer (Radiat. Oncol. Invest.) 90:104-109, 2000. PMID- 10814962 TI - Reply PMID- 10814961 TI - Iodine-125 vs. palladium-103: long-term complications. PMID- 10814963 TI - Notice: recently published book: proton therapy and radiosurgery, H. Breuer, B.J. Smit, springer-verlag; 2000 PMID- 10814964 TI - Allergy to drugs: antioxidant enzymic activities, lipid peroxidation and protein oxidative damage in human blood. AB - Reactive oxygen species lead to lipid peroxidation and specific oxidation of some specific enzymes, proteins and other macromolecules, thus affecting many intra- and intercellular systems. Recently, antioxidant functions have been linked to anti-inflammatory properties. Cell defences against toxic oxygen include antioxidant enzymes. We studied the enzymic antioxidant capacity in human blood of both erythrocytes and mononuclear cells from patients suffering from an allergic reaction to different drugs. We determined superoxide dismutases (SODs), glutathione peroxidase (GSHPx) and catalase (CAT) activities in each cell type. We also determined the extent of thiobarbituric acid reactive substances (TBARS) and the oxidative damage to proteins, in order to study the correlation between the cellular enzymic activities, the oxidative status and the allergic reaction. In mononuclear cells from allergic patients, SODs and CAT activities were enhanced compared with controls. Conversely, a decrease in GSHPx activity was found. In erythrocytes, higher values for CAT, GSHPx and SODs activities were found in allergic patients. TBARS were also enhanced in both types of cells, and the carbonyl content of serum was equally increased. The respective enzymic imbalances in mononuclear cells and erythrocytes, namely, GSHPx/SOD and CAT/SOD, and their consequences are discussed. To our knowledge, this is the first global study of antioxidant enzyme determinations, including TBARS level and carbonyl content, in patients suffering from allergies to drugs. PMID- 10814965 TI - The protective effect of lisinopril on membrane-bound enzymes in myocardial preservation. AB - A number of studies have reported that oxidant stress reduces the activity of isolated Na(+)-K(+) ATPase and Ca(2+) ATPase which are known to affect the cell membrane integrity. The aim of the study is to determine whether the administration of lisinopril is able to protect the membrane-bound enzyme levels in isolated guinea pig hearts and also ascertain whether or not a relationship exists between oxygen free radicals and membrane bound Na(+)-K(+) ATPase and Ca(2+) ATPase. Forty guinea pig hearts were studied in an isolated Krebs Henseleit solution-perfused Langendorff cardiac model. In all groups cardioplegic arrest was achieved by administering St. Thomas' Hospital cardioplegic solution (STHCS). Group 1 (control, n=10) received only STHCS. Group 2 (n=10) were arrested with lisinopril (l micromol l(-1)) added STHCS. Group 3 (n=10) were pretreated with oral lisinopril (0.2 mg kg(-1) twice a day) for 10 days and then arrested with STHCS. Group 4 were also pretreated with oral lisinopril (0.2 mg kg(-1) twice a day for 10 days), arrested with STHCS and reperfused with lisinopril added to Krebs-Henseleit solution (l micromol l(-1)). Hearts were subjected to normothermic global ischaemia for 90 min and then reperfused at 37 degrees C. Pretreatment and addition of lisinopril in the reperfusion buffer improved the levels of membrane-bound enzymes. When the treated groups were compared with control hearts, the best results were achieved in group 4. The Na(+)-K(+) and Ca(2+) ATPase levels increased from 466.38+/-5.99 to 560.12+/ 18.02 and 884.69+/-9.13 to 1287.71+/-13.01 nmolPi mg(-1) protein h(-1) respectively (p<0.05). These results suggest that lisinopril protects the cell membrane integrity and lessens free radical-induced oxidant stress. PMID- 10814966 TI - Inhibitory effect of Cordyceps sinensis and Cordyceps militaris on human glomerular mesangial cell proliferation induced by native LDL. AB - Native LDL, in low concentrations, promotes proliferation of cultured human glomerular mesangial cells. LDL stimulated [(3)H]-thymidine incorporation into DNA of human glomerular mesangial cells. Increased concentrations of LDL led to increased [(3)H]-thymidine incorporation. When LDL concentrations were 5, 10 and 50 microg ml(-1), [(3)H]-thymidine incorporation was 919.5+/-216, 1106+/-132, and 1200+/-210, respectively. When Cordyceps sinensis 100, 200, 300, 400 microg ml( 1) plus LDL 10 microg ml(-1) were added, [(3)H]-thymidine incorporation was 99+/ 19 and 53+/-8, respectively, P<0.01 compared with controls. With Cordyceps militaris at similar concentrations plus LDL 10 microg ml(-1), [(3)H]-thymidine incorporation was respectively 192+/-75, 168+/-66, 145+/-53 and 72+/-16, P<0.01 compared with controls. The data suggest that LDL may play a critical role in mediating mesangial cell hypertrophy or proliferation involved in the development of glomerulosclerosis. Cordyceps sinensis and Cordyceps militaris inhibited, to a certain degree, proliferation of cultured human glomerular mesangial cell induced by LDL. PMID- 10814967 TI - Erythrocyte lipid peroxidation and antioxidants in cigarette smokers. AB - The present study has analysed the relationship between lipid peroxidation and antioxidant status in erythrocytes from 30 adult male cigarette smokers and an equal number of age and sex-matched normal subjects. Erythrocyte lipid peroxidation was markedly increased. The enzymic antioxidants were decreased in erythrocytes of cigarette smokers. The present study highlights the occurrence of lipid peroxidation and possible breakdown of antioxidant status in cigarette smoking. PMID- 10814968 TI - Salivary biomass assessed by bioluminescence ATP assay related to (bacterial and somatic) cell counts. AB - The present work aimed (1) to evaluate ATP content in saliva by the bioluminescent luciferin-luciferase method, (2) to evaluate the relationships between ATP content, bacterial count and epithelial cell numbers in saliva, (3) to study the effect of two different antiseptics (peroxidase system producing hypothiocyanite and chlorhexidine) on the salivary biomass. In 45 young adults, the salivary ATP content ranged from 8 to 1515 nM. Salivary ATP content was significantly and directly correlated to bacterial count and epithelial cell numbers (Spearman-Rank correlation, P< or =0.001). Regression analysis allowed the inference of a mean epithelial cell and bacterial ATP content of 152.7 fg and 8.3 fg per cell, respectively. The salivary ATP content decreased significantly to 38. 8+/-12.3 per cent (mean+/-SEM, N=6) of its initial value after a 30-min incubation in the presence of a peroxidase system producing hypothiocyanite (OSCN(-)). Chlorhexidine (CHX) reduced salivary ATP content to 52.0+/-16.7 per cent. OSCN(-) did not affect the transformed logarithm of bacterial count but CHX reduced it from 7. 02+/-0.26 to 0.52+/-0.33. No effect of OSCN(-) was seen on the ratio of epithelial cell viability while CHX reduced it from 46.7+/-5.1 to 3.9+/ 1.1 per cent. It is concluded that the combination of the evaluations of the ATP content and cell numbers in saliva can provide reliable data about the effects of oral antiseptics on salivary biomass. PMID- 10814969 TI - Effects of antioxidant vitamins A, C, E and trace elements Cu, Se on CuZn SOD, GSH-Px, CAT and LPO levels in chicken erythrocytes. AB - The biologically damaging effects of reactive oxygen species are controlled in vivo by a wide spectrum of antioxidant defence mechanisms. Dietary constituents of antioxidant vitamins and trace elements may play an important role in protecting against oxidant damage. The effects of supplementation of vitamins A, C, E and trace elements Cu and Se on the activities of antioxidant enzymes and lipid peroxide levels in chicken erythrocytes were investigated depend on the time. CuZnSOD activity and plasma Cu levels in the Cu group were increased by 39 and 37 per cent respectively. CuZnSOD activity in vitamin C groups was also increased by 20 per cent. The GSH-Px activity in Se, Se+E and Se+Cu groups was raised by 35, 46 and 69 per cent respectively. Also, the GSH-Px activity in the vitamin C group was increased by 33 per cent. Catalase activity in all of these groups was not significantly different when compared with controls (p<0.01). The maximum decrease in LPO levels of 42 per cent was obtained for the Se+E group. PMID- 10814970 TI - Antimicrobial and biological effects of ipemphos and amphos on bacterial and yeast strains. AB - In this study, the antimicrobial effects of monophosphazenes such as SM ipemphos and amphos were examined on bacterial and yeast strains. In addition, the biological effects of these compounds were tested on the lipid level of Saccharomyces cerevisiae and Candida albicans cells. The SM has an antimicrobial effect on the bacterial and yeast strains within the range of 100 and 1500 microg. When the concentration was increased, the inhibition zone expanded on the growth media ( p < 0.01; p < 0.001). The ipemphos did not affect the bacterial and yeast cells in the 100 and 600 microg range. In addition, the amphos did not show an antimicrobial effect on the bacterial cells between 100 and 300 microg or on yeast cells at any of the administered concentrations. In vitro media, the biological effects of these molecules were compared with vitamin E, melatonin and fish oil on the yeast cells. We have found that monophosphazenes have growth effects on the cells in vitro media. The lipid level of S. cerevisiae cells was decreased by 300 microg doses of vitamin E, fish oil, and ipemphos (respectively; p < 0.05, p < 0.01, and p < 0. 001). In addition, the lipid levels of the same yeast cells were depressed by 1000-microg doses in all supplemented groups. However, it was observed that the highest decrease in lipid level of S. cerevisiae cells occurred in the amphos group ( p < 0.001). The lipid levels of the C. albicans cells were significantly reduced ( p < 0.01) by 300 microg of amphos and melatonin. In contrast, the vitamin E and fish oil significantly raised ( p < 0.01; p < 0.001) the lipid level of the same yeast cell, as compared with the control. In addition, the lipid level of these cells was increased by administration of 1000 microg vitamin E, and melatonin ( p < 0.01). In conclusion, while high concentrations of ipemphos and amphos have an antimicrobial effect on bacterial and yeast cells, amphos did not affect the yeast cells. While ipemphos and amphos increased cell growth in media, they reduced the lipid level of C. albicans and S. cerevisiae. In addition, the antioxidants such as vitamin E, melatonin, and fish oils affected the lipid level of yeast cells. PMID- 10814971 TI - Oxidative metabolism and release of myeloperoxidase from polymorphonuclear leukocytes obtained from blood sedimentation in a Ficoll-Hypaque gradient. AB - Polymorphonuclear neutrophils (PMN) have an important role in the host defence response to infection. These cells produce large amounts of reactive oxygen species (O(2).(-), H(2)O(2) and ONOO(-)) with microbicidal activity. PMN are commonly isolated from peripheral blood by sedimentation through a gradient of density (Ficoll-Hypaque gradient and dextran), yielding a highly homogeneous cellular population. However, some cellular activation due to membrane perturbation is also expected. We studied how the production of reactive oxygen species and release of myeloperoxidase (MPO) from blood PMN are affected by the use of the Ficoll-Hypaque density gradient. PMN isolated by spontaneous sedimentation and total blood were used for comparisons. Lucigenin- and luminol enhanced chemiluminescence was used to estimate the production of reactive oxygen from intact cells and shown to be higher for cells isolated by density gradient both in the absence and presence of added stimuli. The release of MPO, estimated by the chemiluminescence of the luminol/H(2)O(2) reaction in the supernatant of PMN incubated in the absence and presence of stimuli and absence and presence of cytochalasin B, was also higher for PMN isolated by a density gradient. In conclusion, it was shown that the PMN isolation procedure affects reactive oxygen species production and MPO release and in some cases may cause a misinterpretation of results. PMID- 10814972 TI - Phospholipase C and D in the commitment to survival or death in the early lag phase of tetrahymena cultures. AB - We made three kinds of experiments in order to elucidate aspects of physiological mechanisms involved in a series of specific events leading to either cell death or survival in the lag phase of culture growth. We studied the fate of newly inoculated Tetrahymena cells in small droplets at 'high' (more than 1000 cells ml(-1)) and 'low' cell densities (less than 600 cells ml(-1)) in a nutrionally complete, synthetic nutrient medium. Confirming previous results we found that the cells in high-density cultures multiplied to final densities around 500,000 cells ml(-1) and that cells in low-density cultures died before multiplying. The appearance of the cells was recorded with a video camera at 20 frames per second for 6 h or until they died. The results indicated that the death process took place within milliseconds. We also studied the effects of U 73122, an inhibitor of the phosphatidylinositol-specific phospholipase C, on cell survival at low densities. At low inhibitor concentrations low-density cells were rescued from dying. At high inhibitor concentrations all cells died, and phosphatidylinositol but not phosphatidylserine and phosphatidylcholine - saved them. The results indicate that the paths leading to either cell death or to cell proliferation separate within the first few minutes after subcultivation into a new medium, since the first cells in each culture died within 4-30 min after inoculation. Our results also indicate that some PLC activity was required for stimulation of phospholipase D, and that cell death during the early lag phase is caused by a shortage in phosphatidylinositol before the phospholipase D activity is upregulated. These experiments are shedding light on the lethal consequences of a cellular depletion of the important signalling compound phosphatidylinositol in an in vivo system, and may help to elucidate mechanisms behind the century-old fact that eukaryote cells die when inoculated at too low a cell density to survive. PMID- 10814973 TI - Angiotensin converting enzyme gene polymorphism in Alzheimer's disease. PMID- 10814974 TI - Effect of iodide on total antioxidant status of human serum. AB - Free radicals and subsequent lipid peroxidation have been implicated in the pathogenesis of several degenerative and chronic diseases which are also treated frequently in spas. There are some data arising from previous studies which support an antioxidant or scavenging effect of iodide, being the essential ingredient of a therapeutically used local brine. The aim of the study was to test the antioxidant capacity of iodide in human serum. For this reason we measured the so-called Total Antioxidant Status determined by a colorimetric method, which reflects the protection against the attack of reactive oxygen species, including enzymic and non-enzymic antioxidants. Exogenous iodide applied as NaI, shows a significantly increased antioxidant capacity in comparison with NaCl at a concentration of 15 microM, which is quite comparable to the upper range of serum iodide levels achieved through balneo-therapeutical intervention. This result is in accordance with previous results from in vitro depolymerization experiments with hyaluronic acid. The antioxidant effect of 15 microM NaI has been found to be approaching the physiologically relevant concentration of ascorbic acid (50 microM). PMID- 10814975 TI - Developmental changes in capacitative Ca(2+) entry in mouse mammary epithelial cells. AB - Developmental changes in capacitative Ca(2+) entry and Ca(2+) release from intracellular stores were measured using fura-2 fluorescence method during the pregnancy period (day 3-;18) in mouse mammary epithelial cells. Ca(2+) release was identified with the transient intracellular Ca(2+) ([Ca(2+)](i)) increase induced by thapsigargin addition in a Ca(2+)-free solution. Capacitative Ca(2+) entry was measured by the transient [Ca(2+)](i) increase induced by re-addition of extracellular Ca(2+) after depletion of Ca(2+) stores by thapsigargin. The capacitative Ca(2+) entry was greatest at the early stage of pregnancy (i.e. day 3 of pregnancy) and decreased as pregnancy progressed, while Ca(2+) release remained unchanged throughout the developmental stages. These findings indicate that in contrast to Ca(2+) release, a close correlation exists between capacitative Ca(2+) entry and pregnancy-induced development in mammary epithelial cells. PMID- 10814976 TI - Summarizing the goodness of fit of generalized linear models for longitudinal data. AB - This paper extends four goodness-of-fit measures of a generalized linear model (GLM) to random effects and marginal models for longitudinal data. The four measures are the proportional reduction in entropy measure, the proportional reduction in deviance measure, the concordance correlation coefficient and the concordance index. The extended measures satisfy the basic requirements for measures of association. Two examples illustrate their use in model selection. PMID- 10814977 TI - Residuals analysis of the generalized linear models for longitudinal data. AB - The generalized estimation equation (GEE) method, one of the generalized linear models for longitudinal data, has been used widely in medical research. However, the related sensitivity analysis problem has not been explored intensively. One of the possible reasons for this was due to the correlated structure within the same subject. We showed that the conventional residuals plots for model diagnosis in longitudinal data could mislead a researcher into trusting the fitted model. A non-parametric method, named the Wald-Wolfowitz run test, was proposed to check the residuals plots both quantitatively and graphically. The rationale proposedin this paper is well illustrated with two real clinical studies in Taiwan. PMID- 10814978 TI - Improving interpretability: gamma as an alternative to R(2) as a measure of effect size. AB - A traditional measure of effect size associated with tests for difference between two groups is the variance explained by group membership (R(2)). If exposure to a disease causes a small but long term deficit in performance, however, R(2) does not capture that cumulating effect. We propose an alternative statistic, gamma, based on the probability of an unexposed person outperforming an exposed person. Although gamma is also a point estimate, it more easily conveys what the cumulating effect of a deficit would be. We discuss some of the advantages of this measure. PMID- 10814979 TI - A latent class mixed model for analysing biomarker trajectories with irregularly scheduled observations. AB - This paper considers a latent class model to uncover subpopulation structure for both biomarker trajectories and the probability of disease outcome in highly unbalanced longitudinal data. A specific pattern of trajectories can be viewed as a latent class in a finite mixture where membership in latent classes is modelled with a polychotomous logistic regression. The biomarker trajectories within a latent class are described by a linear mixed model with possibly time-dependent covariates and the probabilities of disease outcome are estimated via a class specific model. Thus the method characterizes biomarker trajectory patterns to unveil the relationship between trajectories and outcomes of disease. The coefficients for the model are estimated via a generalized EM (GEM) algorithm, a natural tool to use when latent classes and random coefficients are present. Standard errors of the coefficients are calculated using a parametric bootstrap. The model fitting procedure is illustrated with data from the Nutritional Prevention of Cancer trials; we use prostate specific antigen (PSA) as the biomarker for prostate cancer and the goal is to examine trajectories of PSA serial readings in individual subjects in connection with incidence of prostate cancer. PMID- 10814980 TI - Pros and cons of permutation tests in clinical trials. AB - Hypothesis testing, in which the null hypothesis specifies no difference between treatment groups, is an important tool in the assessment of new medical interventions. For randomized clinical trials, permutation tests that reflect the actual randomization are design-based analyses for such hypotheses. This means that only such design-based permutation tests can ensure internal validity, without which external validity is irrelevant. However, because of the conservatism of permutation tests, the virtues of permutation tests continue to be debated in the literature, and conclusions are generally of the type that permutation tests should always be used or permutation tests should never be used. A better conclusion might be that there are situations in which permutation tests should be used, and other situations in which permutation tests should not be used. This approach opens the door to broader agreement, but begs the obvious question of when to use permutation tests. We consider this issue from a variety of perspectives, and conclude that permutation tests are ideal to study efficacy in a randomized clinical trial which compares, in a heterogeneous patient population, two or more treatments, each of which may be most effective in some patients, when the primary analysis does not adjust for covariates. We propose the p-value interval as a novel measure of the conservatism of a permutation test that can be defined independently of the significance level. This p-value interval can be used to ensure that the permutation test have both good global power and an acceptable degree of conservatism. PMID- 10814981 TI - Non-parametric test of ordered alternatives in incomplete blocks. AB - Often in medical studies, study subjects become a natural block of observations repeated over a time period. Some subjects miss observations, thus becoming incomplete blocks of observations. We are interested in testing an ordered alternative (or time trend), and propose a non-parametric method to detect a trend in incomplete blocks. Our approach is to estimate the trend by the linear regression method within each block and apply the one-sample Wilcoxon test to the estimated linear trends. The one-sample Wilcoxon test will be sensitive to the trend if it exists. The proposed test statistic is asymptotically normal and consistent. We can also estimate the overall magnitude of the linear trend and its confidence interval by a proper non-parametric method. By Monte Carlo studies, we compare the performance of the proposed test against extended Page and Jonckheere tests. Published in 2000 by John Wiley & Sons, Ltd. PMID- 10814982 TI - A quality control measure for longitudinal studies with continuous outcomes. AB - At the time of data acquisition in a longitudinal study a decision needs to be made whether or not the latest measurement of the primary outcome is a potential outlier. If the data point does not fit with the subject's prior data, the patient can be immediately remeasured before he/she leaves the office. From the third visit onwards, a least squares approach can be used to generate prediction intervals for the value of the response at that visit. We propose a Bayesian method for calculating a prediction interval that can incorporate external information about the process that can be used beginning at the first visit. Both the least squares and Bayesian approaches will be used to prospectively clean longitudinal data. An example using longitudinally measured bone density measurements in the elderly will be discussed. In addition, simulation studies will be described which show that both cleaning methods are better than doing nothing and that the Bayesian approach outperforms the least squares method. PMID- 10814983 TI - A comparison of three tests to detect general clustering of a rare disease in Santa Clara County, California. AB - Statistical tests to detect clustering of a rare disease investigate whether an observed spatial pattern of cases appears to be due to chance alone. Heterogeneous population density and the geographic structure of the data under consideration complicate the ability to make comparisons of different tests. Further, interpretation of test results depends on the nature of the test used and what feature of the data it is designed to detect. With these issues in mind, we compare three recent tests for assessing general clustering among cases where the population is distributed heterogeneously across the study area, namely those of Besag and Newell, Turnbull et al. and Tango. We compare these methods using 1981 incidence data for severe cardiac birth defects from Santa Clara County, California. PMID- 10814984 TI - Titus H.J. Huisman, M.D., ph.D PMID- 10814985 TI - Determination of beta-globin gene cluster haplotypes and prevalence of alpha thalassemia in sickle cell anemia patients in Venezuela. AB - Sickle cell anemia and alpha-thalassemia have a heterogeneous distribution in Venezuela with a high frequency in the coastal area (sea level) and few cases in the mountains. Most of our population is an ethnic admixture of Europeans (Spaniards colonists), Africans (slaves), and Amerindians. The purpose of our study was to determine the origin of the beta(s) globin haplotype, age and survival dependency, and the admixture among the different African groups in our population. The alpha(3.7) globin gene deletion status was also studied and found in a very high frequency. DNA from peripheral blood of 191 non-related patients (81 with HbS homozygous and 15 patients compound heterozygous for HbS, HbC, HbD with beta-thalassemia, and 95 with sickle cell trait) were studied. The beta(s) chromosome was linked 51% to the Benin Haplotype, 29.5% with the CAR, 12.5% to the Senegal, and 2.5% to the Cameroon. We did not find any significant difference between the haplotype distribution among adults and children and among sickle cell patients and traits. Only 8.6% of the patients have homozygosity for the Benin haplotype. These results show a very high frequency of admixture in our African origin population. PMID- 10814986 TI - Tumor necrosis factor-alpha is undetectable in the plasma of SS patients with elevated Hb F. AB - Steady-state sickle cell disease (SCD) patients may have increased plasma levels of acute phase reactants and pro-inflammatory cytokines because of subclinical inflammation. We have estimated TNF-alpha levels in the plasma and in supernatants following peripheral blood mononuclear cell (PBMC) activation with phytohemagglutinin (PHA) in a group of Kuwaiti SCD patients using ELISA. The group consisted of 28 SS, 8 Sbeta-thal, and 2 SD patients all in steady state; 5 SS patients were studied during 7 episodes of painful crisis. The subjects were aged 2 to 16 years, with a mean of 7.3 +/- 3.5 years. The beta(S)-globin gene cluster haplotype, alpha-tha1 status, and spleen function were determined in the SS group using standard techniques. Most (82%) were homozygous for the Saudi Arabia/India haplotype and had elevated Hb F levels ranging from 15% to 35%. There were 24 controls (Hb AA or AS), of whom 14 were healthy and 10 were acutely ill at the time of the study. None of the children with SCD (either in steady state or crisis) had detectable plasma TNF-alpha, but four controls (3 acutely ill and one healthy) had levels ranging from 61.7 to 249.8 pg/mL. Following PHA stimulation most subjects responded with high levels of TNF-alpha, with the median level among the steady-state SS patients being significantly higher than that in the controls (both the acutely ill or healthy). It therefore appears that because of the mild disease among our Arab SS children, TNF-alpha is not detectable in their plasma in steady state; these children, however, had a significantly higher response than controls following PBMC activation. PMID- 10814987 TI - Hepatitis C virus in non-Hodgkin's lymphoma. A reappraisal after a prospective case-control study of 300 patients. Lombart Study Group of HCV-Lymphoma. AB - It is widely thought, but not yet explained, that there might be a pathogenetic link between the infection of hepatitis C virus (HCV) and the onset of B non Hodgkin's lymphoma (NHL). We studied the prevalence of serum anti-HCV antibodies among 300 NHL comparing it with the prevalence among 600 age- and sex-matched non neoplastic subjects as controls, 247 patients with non-lymphomatous neoplasm, and 122 patients treated with immunosuppressive agents. We found a prevalence of 0.16 among NHL and 0.085 among controls and non-lymphomatous patients. Although the difference was statistically significant (P < 0.001), the odds ratio was 2.049 and its confidence intervals included the equality. The HCV prevalence was independent of NHL subset, and the genotypes distribution was the same among NHL and controls. We disclosed a HBsAg prevalence of 0.077 in NHL versus 0.008 in controls (P < 0.001) with an odds ratio of 9.9. We do not believe that these findings support the hypothesis of an HCV pathogenetic role in lymphomagenesis because (i) the risk of previous infection is marginally higher in NHL than in controls, (ii) a typical genotype distribution is lacking, as is a NHL clinico histological feature associated with HCV, and (iii) the higher prevalence of viral infection is not specific as witnessed by the high HBsAg prevalence. PMID- 10814988 TI - Quantitation of minimal residual disease in t(8;21)-positive acute myelogenous leukemia patients using real-time quantitative RT-PCR. AB - t(8;21) is one of the common chromosomal translocations in acute myelogenous leukemia (AML). Using a recently developed real-time quantitative polymerase chain reaction (PCR) system, we analyzed the minimal residual disease (MRD) in bone marrow samples from seven AML patients with t(8;21) at different time points during the clinical courses of their disease. Four of these patients received chemotherapy and allogenic bone marrow transplantation (allo-BMT), and the other three were treated with chemotherapy alone. Two of the patients that received allo-BMT suffered a relapse. In these patients, the levels of AML1-MTG8 mRNA expression were shown to quantitatively increase. After re-induction chemotherapy and donor lymphocyte infusion therapy, AML went into remission and the expression levels decreased. In the other two patients receiving allo-BMT, the disease went into remission and the level of AML1-MTG8 mRNA expression remained under the detectable range. The other three patients received several courses of chemotherapy, without allo-BMT, and all of them clinically reached the hematological and cytogenetic remission state. However, there were low but detectable levels of MRD in their bone marrow samples. These results suggest that the real-time quantitative PCR assay is very useful for the monitoring of MRD and detecting an early relapse. This assay may also be useful in determining the quantitative difference in myelo-ablative activity between the chemotherapy alone and chemotherapy in conjunction with allo-BMT. PMID- 10814990 TI - Hemostatic effects of low-dose protamine following cardiopulmonary bypass. AB - Twenty-eight patients undergoing cardiac surgery were prospectively studied and were assigned to two groups. The patients received 0.8- (Group L) or 2.0-fold (Group H) dose of protamine for the neutralization after cardiopulmonary bypass (CPB) which was determined by Hepcon HMS(R) assay system in which the reagent chamber containing the concentration of protamine that completely neutralized the heparin had the shortest clotting time. Mean dose of protamine was 1.60 +/- 0.50 mg kg(-1) in Group L, and 3.56 +/- 1.48 mg kg(-1), respectively. Activated clotting times (ACT) were comparable between the two groups through this study period. In Group H, platelet counts significantly decreased to 69% of that before protamine administration, and plasma platelet factor 4 level significantly increased to approximate 2-fold of that before protamine administration just after protamine administration, respectively. However, these phenomena were not observed in Group L. In addition, these hemostatic changes occurred transiently just after protamine administration. We conclude that the low-dose protamine may prevent transient platelet depletion following CPB. Low-dose protamine can neutralize anticoagulation effect of heparin sufficiently and may mitigate protamine-induced platelet dysfunction. PMID- 10814989 TI - Hepatitis C virus infection and lymphoproliferative diseases in France: a national study. The GERMIVIC Group. AB - The putative role of hepatitis C virus (HCV) infection in the pathophysiology of lymphoproliferative diseases (LPD) is supported by North American and southern European studies reporting high HCV seroprevalence in patients with B-cell-non Hodgkin lymphoma (NHL). In order to evaluate the situation in France, we conducted a retrospective national study about the association of chronic HCV infection and LPD. 72 Internal Medicine and Infectious Diseases departments were contacted. Response rate was 51.4%. We recorded 43 LPD (19 males, 24 females): 31 B-cell-NHL, 4 Waldenstrom's macroglobulinemia, 3 chronic lymphocytic leukemia, 2 multiple myeloma, 2 lymphomas of the mucosa-associated lymphoid tissue, and 1 Hodgkin's disease. Mean age at HCV diagnosis was 62 years (range 33-84). In 16 cases, LPD occurred in patients known to be HCV-infected. For 11 patients, LPD diagnosis preceded the diagnosis of HCV infection, whereas diagnosis was done simultaneously in 11 patients. For those with accurate infection date, mean interval between both events was 15.2 years. Fourteen patients had HCV extrahepatic manifestations: 9 mixed cryoglobulinemia, including 7 with NHL, 5 sicca syndrome (5 NHL), and both in one patient. Cohort of HCV-infected patients could be accurately determined for 16 departments, totaling 1,485 patients and 37 cases. Thus, from our data the frequency of LPD among HCV-infected patients approximates 2. 49%. Despite possible bias inherent to this retrospective study, our data support the hypothesis of HCV-associated LPD and particularly B-cell NHL. In France, this association is much lower than in Italy. Further studies are needed to assess the precise role of HCV in the multistep process leading to monoclonal proliferation. PMID- 10814991 TI - Elevation of soluble Fas and soluble Fas ligand in reactive macrophage activation syndromes. AB - Derailed T-cell activation can give rise to life-threatening macrophage activation, the final common pathway of the different forms of reactive macrophage activation syndromes (rMAS). Besides inappropriate activation of the immune system, impaired termination of immune responses might be another mechanism leading to rMAS. The Fas (CD95)/Fas ligand (CD95 ligand) system functions in turning off immune responses by executing activation-induced cell death (AICD). Soluble Fas (sFas) and Fas ligand (sFasL) can interfere with their corresponding membrane-bound counterparts, qualifying them as potential parameters of impaired immune termination. Hence, sFas and sFasL were analyzed in sera of rMAS patients. We show that soluble Fas/CD95 (sFas) is elevated >2 SD over the mean of controls in all 8 rMAS episodes studied (mean 12.08 +/- 6.12 ng/mL, range 3.7-20.2; controls 2.46 +/- 0.49, range 1.5-2.9). sFasL was detected during five rMAS episodes (0.70 +/- 0.49 ng/mL, range 0.16-1.28; controls all below the limit of detection of 0.1). In addition, both parameters decrease during convalescence, reflecting clinical evolution. In conclusion, sFas seems to be consistently elevated during acute rMAS. sFasL is detected only in a subgroup of our adult rMAS patients extending the recent finding of sFasL elevation in a majority of children with macrophage activation syndromes (Hasegawa et al. Blood 1998;91(8):2793-2799). By interfering with AICD, sFas and sFasL might contribute to the pathogenesis of at least a subset of rMAS. PMID- 10814992 TI - 5q- syndrome presenting chronic myeloproliferative disorders-like manifestation: a case report. AB - A 28-year-old Japanese woman with suspected essential thrombocythemia (ET) had marked thrombocytosis, mild leukocytosis with normal neutrophil alkaline phosphatase activity, and no anemia. She was monitored without being given any medication. Eleven years later, complete blood counts showed no remarkable changes but some non-lobulated mononuclear megakaryocytes were found in the bone marrow. Cytogenetic analysis revealed deletion of the long arm of chromosome 5 (5q-). Subsequently, hemoglobin and platelet counts decreased gradually, splenomegaly appeared and progressed, after which myelofibrosis developed. Acute leukemia developed 16 years after the first documentation of thrombocytosis. 5q- syndrome is known to be a myelodysplastic syndrome (MDS) with unique clinical features and cases with this syndrome presenting with thrombocytosis of more than 1,000 x 10(9)/L but without anemia are rare. Furthermore, it is noteworthy that in this patient transition to acute leukemia occurred following development of myelofibrosis and marked splenomegaly, which are generally observed in blastic crises resulting from chronic myeloproliferative disorders (CMPD). The patient showed features indicative of CMPD rather than of MDS in spite of presenting with 5q- chromosomal abnormality. This case supports the concept of "mixed myelodysplastic and myeloproliferative syndromes" and suggests the possibility of the appearance of CMPD-like manifestations in 5q- syndrome. PMID- 10814993 TI - TCR-Vbeta repertoire analysis with RT-PCR was useful for the early detection of pulmonary relapsed T-cell lymphoma after autologous peripheral blood stem cell transplantation. AB - Pulmonary recurrence of malignant lymphoma is a rare event after stem cell transplantation. We report here a 45-year-old male who was successfully diagnosed with relapsed pulmonary T-cell lymphoma using an RT-PCR method. Clonal expansion of T cells expressing identical TCR V-D-J junction size (Vbeta5-Jbeta1.5) was demonstrated in lymphocyte groups obtained from both bronchoalveolar lavage fluid at relapse, and paraffin embedded lymph node samples resected when he was first diagnosed with angioimmunoblastic T-cell lymphoma. This method provided evidence to diagnose relapsed pulmonary angioimmunoblastic T-cell lymphoma in its early phase. PMID- 10814994 TI - Large deletion of the X-linked lymphoproliferative disease gene detected by fluorescence in situ hybridization. AB - The X-linked lymphoproliferative disease (XLP) is an inherited immunodeficiency characterized by an abnormal responses to infection with Epstein-Barr virus (EBV), resulting in fatal infectious mononucleosis, hypogammaglobulinemia, virus associated hemophagocytic syndrome, and malignant lymphoma. Mutations in the gene coding for a T cell-specific SLAM-associated protein (SAP) have been recently identified in XLP patients. We report on a 1-year-old boy representing fulminant hemophagocytic syndrome. He developed high fever, lymphadenopathy, hepatosplenomegaly with liver dysfunction, and pancytopenia with marrow hemophagocytosis. EBV DNA was abnormally increased in the blood. Polymerase chain reaction failed to amplify SAP mRNA and genomic DNA products from the patient' As peripheral blood. A large deletion of the SAP gene was confirmed by fluorescence in situ hybridization (FISH). FISH analysis also disclosed that the patient's mother was a carrier. We conclude that FISH can be useful in the diagnosis of XLP with large deletions of the SAP gene and its carrier state. PMID- 10814995 TI - Two cases of chronic graft-versus-host disease with elevated levels of soluble Fas ligand in serum. AB - It has recently been shown that the Fas-Fas ligand (FasL) system may be one of the pathogeneses for acute graft-versus-host disease (GVHD), and it has been reported that serum soluble Fas ligand (sFasL) increases with the presence of acute GVHD. However, there is no report on a correlation between the Fas-FasL system and chronic GVHD. We present two cases of chronic GVHD with elevated levels of serum sFasL. Its level in each case was high at the onset of chronic GVHD, but it decreased with steroid therapy. Liver dysfunction also improved as the level of serum sFasL decreased. It appears in these cases that the Fas-FasL system was related to the pathogenesis of liver damage. PMID- 10814996 TI - Acute brachial artery thrombosis as the initial manifestation of human immunodeficiency virus infection. AB - Thrombosis of upper extremity arteries is most commonly due to atherosclerosis of the proximal subclavian artery, trauma, or catheter-related injury. In the absence of an identifiable cause, a search for a hypercoagulable state is indicated. Hematologic manifestations of human immunodeficiency virus (HIV) infection and AIDS are frequent occurrences (Coyle TE. Med Clin N Am 1997;81:449 476). The most important of these are cytopenias (anemia, neutropenia, and thrombocytopenia). The incidence and severity of cytopenia are generally correlated to the stage of the HIV infection. In addition, various coagulation abnormalities have been reported in HIV-infected patients. Apart from thrombocytopenia, these have included a prolonged APTT due to the presence of lupus anticoagulant, an increased prevalence of protein S and heparin cofactor II deficiency, and hypoalbuminemia-related fibrin polymerization defects (Toulon P. Ann Bio Clin (Paris) 1998;56:153-160). HIV infection has also been associated with endothelial dysfunction. Although for the most part asymptomatic, elevated D dimer levels have been found in HIV-infected patients, suggesting the existence of a prethrombotic state. In fact, clinical thrombosis eventuates in 2% of these patients (Toulon, 1988). Documented thromboses have involved both veins and arteries. We hereby present a patient who developed an acute thrombosis of his brachial artery as the initial manifestation of HIV infection. PMID- 10814997 TI - Life-threatening splenic hemorrhage in two patients with Gaucher disease. AB - Massive splenomegaly is a frequent finding in patients with Gaucher disease, the most common of the sphingolipidoses. Even so, the risk for splenic rupture and intracapsular hemorrhage has not been emphasized due to the rarity of this occurrence and the fibrotic, rubbery consistency of splenic tissue in these patients. We report two adult patients with type 1 Gaucher disease who suffered life-threatening splenic bleeds that were not acutely diagnosed. Both patients ultimately required emergent splenectomies. Factors complicating the diagnosis of splenic hemorrhage in patients with Gaucher disease are discussed. Published 2000 Wiley-Liss, Inc. PMID- 10814998 TI - Arterial thrombosis, GM-CSF, and the lupus anticoagulant. PMID- 10814999 TI - Posttranslationally modified tubulins and microtubule organization in hemocytes of the brine shrimp, Artemia franciscana. AB - Crustaceans possess blood cells (hemocytes) that mediate organismal defense and are analogous to vertebrate leukocytes. In order to more fully characterize these types of cells, hemocytes of the branchiopod crustacean, Artemia franciscana, were analyzed. The data indicate that Artemia have one type of hemocyte, ranging in morphology from compact and spherical to flat and spreading when examined in vitro. Electron microscopy revealed many cytoplasmic granules in the hemocytes and only a limited number of other membrane-bound organelles. Centrioles and microtubules were also visible in thin sections of chemically fixed samples. The cytoplasm of spherical hemocytes was completely labeled by general antitubulin antibodies, but in flattened hemocytes packing of cytoskeletal elements was less tight and individual microtubules were observed. Probing of Western blots disclosed acetylated, tyrosinated, and detyrosinated tubulin isoforms in hemocyte homogenates, the first characterization of posttranslationally modified tubulins in this cell type. Acetylated tubulin was restricted to a subset of microtubules, whereas tyrosinated microtubules were displayed more abundantly. Staining obtained with antibody to detyrosinated tubulin was unusual because it was limited to the perinuclear region of hemocytes. Incubation of blood cells with a monoclonal antibody to gamma-tubulin yielded fluorescent dots sometimes in pairs, a pattern characteristic of centrosomes. The findings support the conclusion that Artemia hemocytes undergo rapid morphogenesis in vitro accompanied by extensive rearrangement of their microtubules, the latter probably indicative of cytoskeletal changes that occur during cell movement and phagocytosis. Additionally, the hemocytes contain posttranslationally modified alpha-tubulins and centrosome-associated gamma-tubulin, both with the potential to influence microtubule organization and function. PMID- 10815000 TI - Development of the hypochord and dorsal aorta in the zebrafish embryo (Danio rerio). AB - The hypochord of the zebrafish embryo (Danio rerio) emerges at the 9-somite stage as a single row of cells in the dorsomedial endoderm immediately ventral to the notochord. It is recognizable from the 2(nd) or 3(rd) somite and extends along the trunk to the same extent as the somites. A basal lamina surrounds the hypochord and its cells are slightly larger than the nearby endoderm cells. TEM studies have shown that the hypochord cells contain, in addition to mitochondria, well-developed rough endoplasmic reticula and Golgi networks, indicating synthetic activity. Once formed, the hypochord will stay in close association with the notochord, and this axial complex gradually moves dorsally, separating the hypochord from the endoderm as a one-cell-wide, rod-like structure that is bean-shaped in transverse section. This is the situation in the 15-somite embryo, at the level of the 4-5(th) somites. In the gap between the hypochord and the endoderm, angioblast cells aggregate and start to form the dorsal aorta, which becomes intimately associated with the hypochord. In the 17-somite embryo the aortic rudiment is established just ventral to the hypochord as a tube with a lumen. As development proceeds, the size of the hypochord decreases. In the pec fin embryo the hypochord is still recognizable in the posterior trunk, but has apparently vanished in anterior regions. The temporal correlation between the appearance of the hypochord and the formation of the dorsal aorta, coupled with the intimate relationship between these structures, suggest that the hypochord may play a role in the positioning of the dorsal aorta. PMID- 10815001 TI - Ultrastructure of the midgut and hindgut of Derocheilocaris remanei (Crustacea, Mystacocarida). AB - Ultrastructural features and structure of the midgut and hindgut of Derocheilocaris remanei were studied. The large endodermal midgut is differentiated into an anterior midgut and a posterior midgut separated by a conspicuous constriction. Both circular and longitudinal striated muscle bands surround the midgut, while the hindgut only presents longitudinal muscles. The limit between the midgut and the cuticle-lined hindgut is marked by a rectal valve. In cross-section, the short hindgut is triradiate and has a distinct Y shaped lumen. The hindgut cuticular lining appears interrupted at the tip of every branch of the Y. Three different cell types are found in the midgut epithelium: basally located undifferentiated cells that give rise to the other two specialized cell types; secretory zymogen-like cells responsible for extracellular digestion and located mainly in the anterior midgut; and vacuolated cells, distributed all along the midgut and appearing to have several functions, including absorption, intracellular digestion, and nutrient transport. A single basic cell type forms the hindgut epithelium. The suggested function for the hindgut is the transport and ejection of waste products. PMID- 10815002 TI - Ultrastructural study of the boar seminiferous epithelium: changes in cryptorchidism. AB - The present study compares the ultrastructural features of Sertoli cells and germ cells between scrotal testes of healthy boars and abdominal testes of unilateral and bilateral cryptorchid boars. In healthy boars, spermatogonia are flat cells lying in close association with the basal lamina. As differentiation progresses, spermatogonia acquire an oval profile and lose their contact with the basal lamina. Spermatocytes are round cells moving from the basal compartment of the seminiferous epithelium to the luminal compartment. Spermatids exhibit complex morphological changes leading to the formation of spermatozoa. Sertoli cells extend from the basal lamina to the tubular lumen. The nucleus encloses fine euchromatin and one or two nucleoli; the nuclear envelope has a few deep infoldings. The lateral cell membranes form junctional specializations that constitute the blood-testis barrier. The cytoplasm encloses smooth endoplasmic reticulum, vesicles, aggregates, and scattered mitochondria. The seminiferous epithelium of abdominal testes from unilateral and bilateral cryptorchid boars contains few spermatogonia with an abnormal appearance; the alteration in germ cell number is more severe in the bilateral disease. In unilateral cryptorchid boars, spermatogonia appear as either large pyramidal cells or roundish cells; in bilateral cryptorchid boars, spermatogonia show roundish profiles and degenerative patterns. Abdominal testes of both unilateral and bilateral cryptorchid boars are constituted by immature Sertoli cells that show abnormal cytoplasmic content, defective development of the blood-testis barrier, and atypical nuclear appearance; in bilateral cryptorchid boars, immature Sertoli cells exhibit degenerative signs. At postpubertal age, unilateral and bilateral cryptorchidism induce total arrest of spermatogenesis at spermatogonial stage as a result of an abnormal differentiation of the Sertoli cells. Moreover, the degeneration of abdominal testes initiates earlier in bilateral cryptorchidism than in unilateral cryptorchidism. PMID- 10815003 TI - Precocial development of axial locomotor muscle in bottlenose dolphins (Tursiops truncatus). AB - At birth, the locomotor muscles of precocial, terrestrial mammals are similar to those of adults in both mass, as a percent of total body mass, and fiber-type composition. It is hypothesized that bottlenose dolphins (Tursiops truncatus), marine mammals that swim from the instant of birth, will also exhibit precocial development of locomotor muscles. Body mass data from neonatal and adult dolphins are used to calculate Grand's (1992) Neural and Muscular Indices of Development. Using these indices, the bottlenose dolphin is a Condition "3.5" neonate, where Condition 4 is the documented extreme of precocial development in terrestrial mammals. Moreover, myosin ATPase (alkaline preincubation) analyses of the epaxial locomotor m. extensor caudae lateralis show that neonatal dolphins have fiber type profiles very similar to those of adults. Thus, based on mass and myosin ATPase activity, muscle development in dolphins is precocial. However, succinic dehydrogenase and Nile red histochemistry demonstrate that neonatal dolphin muscle has mitochondrial and lipid distributions different from those found in adults. These data suggest that neonates have a lower aerobic capacity than adults. Dolphin neonates may compensate for an apparent lack of aerobic stamina in two ways: 1) by being positively buoyant, with a relatively increased investment of their total body mass in blubber, and 2) by "free-riding" off their mothers. This study investigates quantitatively the development of a dolphin locomotor muscle and offers suggestions about adaptations required for a completely aquatic existence. PMID- 10815004 TI - Effects of saffron extract and its constituent crocin on learning behaviour and long-term potentiation. AB - Crocus sativus L., commonly known as saffron, is used in folk medicine for various purposes. Modern pharmacological studies have demonstrated that saffron extracts have antitumour effects, radical scavenger properties or hypolipaemic effects. Among the constituents of saffron extract, crocetin is mainly responsible for these pharmacological activities. In addition, recent behavioural and electrophysiological studies have demonstrated that saffron extract affects learning and memory in experimental animals. Saffron extract improved ethanol induced impairments of learning behaviours in mice, and prevented ethanol-induced inhibition of hippocampal long-term potentiation, a form of activity-dependent synaptic plasticity that may underly learning and memory. This effect of saffron extract is attributed to crocin (crocetin di-gentiobiose ester), but not crocetin. Saffron extract or its active constituents, crocetin and crocin, could be useful as a treatment for neurodegenerative disorders accompanying memory impairment. PMID- 10815005 TI - Effect of selected thallophytic glucans on learning behaviour and short-term potentiation. AB - This paper reviews the effects of thallophytic glucans on rodent cognitive performance modelled by a combination of behavioural and electrophysiological approaches. Glucans were isolated from thallophytic plants, based on prescriptions used in traditional Chinese and Japanese medicine. In parallel with the already described enhancement of hippocampal synaptic plasticity by disaccharides, polysaccharides isolated from lichens Flavoparmelia caperata and Cetrariella islandica, enhanced hippocampal plasticity and behavioural performance in rats. PMID- 10815006 TI - Constituents of Croton menthodorus and their effects on electrically induced contractions of the guinea-pig isolated ileum. AB - The present study examines the effects of the extracts [petroleum ether, CHCl(3), CHCl(3)MeOH (9:1) and MeOH], partially purified fractions and pure compounds from Croton menthodorus on the electrically induced contractions of the isolated guinea-pig ileum (ECI). The results of the experiments indicate that CHCl(3)/MeOH (9:1) and MeOH extracts, tested at concentrations of 100, 50 and 25 microg/mL, dose-dependently reduced the guinea-pig ileum contractions, whereas petroleum ether and CHCl(3) extracts did not affect it. Furthermore, the partially purified fractions III-VI from the CHCl(3)/MeOH extract, each tested at concentrations of 100, 50 and 25 microg/mL also inhibited ECI. Finally, pure compound 1 (6 x 10( 6), 3 x 10(-6), 1 x 10(-6) M) isolated and purified from the most active fraction III significantly reduced, in a dose-dependent manner, the electrical contractions of the ileum. Compound 1 was identified by NMR and EI-MS data as the morphinandien-7-one, O-methylflavinantine. PMID- 10815007 TI - Dihydroquercetin as a means of antioxidative defence in rats with tetrachloromethane hepatitis. AB - The antioxidant effect of dihydroquercetin (DHQ) was studied in Wistar rats with experimental hepatitis, caused by tetrachloromethane (CCl(4)). Animals were divided into three groups: intact (n = 9); control (n = 9) which received CCl(4) subcutaneously for 4 days (4 mL/kg); and experimental (n = 9) which received DHQ (100 mg/kg) for 4 days prior to the first administration of CCl(4) and during the course of the subsequent 14 days. DHQ was intubated per os, using a water crystalline suspension. The content of products of lipid peroxidation, reacting with thiobarbituric acid in the serum and liver of the control animals, was increased by more than 1.5 fold compared with the intact and experimental animals (p < 0.01). The blood plasma antioxidant activity of the control animals was 1.8 to 2 times lower than that of experimental and intact animals (p < 0.01) It is suggested that the data obtained are dependent on the anti-oxidant properties of DHQ. PMID- 10815008 TI - Ursolic acid isolated from Eucalyptus tereticornis protects against ethanol toxicity in isolated rat hepatocytes. AB - Ursolic acid is the active material isolated from the leaves of the Eucalyptus hybrid E. tereticornis. In the present study, it has shown a significant preventive effect in vitro against ethanol-induced toxicity in isolated rat hepatocytes. Compared with the incubation of isolated hepatocytes with ethanol only, the simultaneous presence of ursolic acid in the cell suspension preserved the viability of hepatocytes and reversed the ethanol-induced loss in the level of all the marker enzymes (AST, ALT and AP) studied. Ethanol alone resulted in a 48%-54% decrease in the viability and a 42%-54% reduction in the biochemical parameters of the hepatocytes. Ursolic acid showed a concentration dependent (1 100 microg/mL) preventive effect (12%-76%) on alcohol-induced hepatocyte toxicity by restoring the altered parameters. The results thus suggest the effective use of an in vitro test system as an alternative for in vivo assessment of hepatoprotective activity of purified material. PMID- 10815009 TI - Nutritional supplementation with Chlorella pyrenoidosa for patients with fibromyalgia syndrome: a pilot study. AB - Fibromyalgia syndrome is a common, chronic musculoskeletal disorder of unknown aetiology. While available therapy is often disappointing, most patients can be helped with a combination of medication, exercise and maintenance of a regular sleep schedule. The objective of the present study was to determine if adding nutritional supplements derived from the unicellular green alga, Chlorella pyrenoidosa, produced any improvements in the clinical and functional status in patients with moderately severe symptoms of fibromyalgia syndrome. Eligible patients had 2+ palpable tenderness at 11 or more of 18 defined tender points and had a tender point index (TPI) of at least 22. Each day for 2 months, participants consumed two commercially available Chlorella-based products, 10 g of 'Sun Chlorella' tablets and 100 mL of liquid 'Wakasa Gold'. Any amelioration of symptoms was validated and quantified using semi-objective and subjective outcome measures systematically administered at clinic visits on days 0, 30 and 60 of the diet therapy. Eighteen of the 20 patients enrolled completed the 2 month trial. The average TPI for the group which at onset was 32, decreased to a mean of 25 after 2 months. This decrease was statistically significant (p = 0.01), representing a 22% decrease in pain intensity. Blood samples taken on each occasion indicated no significant alterations in serum chemistries, formed elements, and circulating lymphocyte subsets. Compilations of the results of patient interviews and self-assessment questionnaires revealed that seven patients felt that the dietary supplement had improved their fibromyalgia symptoms, while six thought they had experienced no change, and five believed the symptoms had worsened over the time of the trial. The results of this pilot study suggest that dietary Chlorella supplementation may help relieve the symptoms of fibromyalgia in some patients and that a larger, more comprehensive double-blind, placebo-controlled clinical trial in these patients is warranted. PMID- 10815010 TI - Antioxidant activity of Bacopa monniera in rat frontal cortex, striatum and hippocampus. AB - The effect of a standardized extract of Bacopa monniera Linn. was assessed on rat brain frontal cortical, striatal and hippocampal superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities, following administration for 7, 14 or 21 days. The effects induced by this extract (bacoside A content 82% +/- 0.5%), administered in doses of 5 and 10 mg/kg, orally, were compared with the effects induced by (-) deprenyl (2 mg/kg, p. o.) administered for the same time periods. Bacopa monniera (BM) induced a dose related increase in SOD, CAT and GPX activities, in all the brain regions investigated, after 14 and 21 days of drug administration. On the contrary, deprenyl induced an increase in SOD, CAT and GPX activities in the frontal cortex and striatum, but not in the hippocampus, after treatment for 14 or 21 days. The results suggest that BM, like deprenyl, exhibits a significant antioxidant effect after subchronic administration which, unlike the latter, extends to the hippocampus as well. The results suggest that the increase in oxidative free radical scavenging activity by BM may explain, at least in part, the cognition- facilitating action of BM, recorded in Ayurvedic texts, and demonstrated experimentally and clinically. PMID- 10815011 TI - Hepatoprotective effects of Inula britannica on hepatic injury in mice. AB - Inula britannica, a Kampo medicine, is prepared from the heads of Compositae plants such as Inula britannica L., which has been used clinically as a remedy for nausea, hiccup and excessive sputum. Here it is shown that administration of Inula britannica improves the survival rate of mice with hepatic injury induced by LPS/PA. It is also suggested that administration of Inula britannica significantly reduces the fluctuation in the amount of cytokine in the spleen of mice with hepatic injuries, and that the Th1/Th2 control effect is related to the inhibitory action of Inula britannica against hepatic injury. In vitro testing suggests that Inula britannica suppresses Th1 differentiation and induces Th2 differentiation by inhibiting the production of macrophage IL-12 and promoting the production of IL-10, thus showing the immunological effect of hepatic injury inhibition by affecting the balance between Th1 and Th2. PMID- 10815012 TI - Antiproliferative effects of compounds derived from plants of Northeast Brazil. AB - Ten compounds derived from plants indigenous to Northeast Brazil were examined for antiproliferative effects on human cells in vitro. The effects of these phytochemicals on cell growth were determined by the MTT microtitre assay with 3 day continuous drug exposure. Three human cell lines were used: CEM leukaemia, SW1573 lung tumour and CCD922 normal skin fibroblasts. Four active compounds were found with IC(50) values less than 10 microg/mL in the two cancer cell lines. Oncocalyxones A and C, both 1,4-anthracenediones from Auxemma oncocalyx (Boraginaceae), showed cytotoxicity with mean IC(50) values of 0.8-2, 7-8 and 12 13 microg/mL against CEM, SW1573 and CCD922, respectively. One diterpene and one flavonoid, both from Egletes viscosa (Compositae), were also active. 12-Acetoxy hawtriwaic acid lactone was cytotoxic with mean IC(50) values of 6, 10 and 10 microg/mL, respectively. 4,5-Dihydroxy-3,3,7, 8-tetramethoxy flavone (ternatin) was only growth-inhibitory with mean IC(50) values of 2, 1 and 10 microg/mL, respectively. These four most active compounds were examined further for their effects on DNA integrity and on DNA synthesis. All but ternatin caused substantial DNA damage and marked inhibition of 5-bromo-2'-deoxyuridine incorporation within 24 h. This study demonstrated the antiproliferative activity of four novel phytochemicals, three of which are DNA-reactive and inhibit DNA synthesis. Further studies are warranted to evaluate these compounds for antitumour potential. PMID- 10815013 TI - Inhibitory effect of Byakko-ka-ninjin-to on itch in a mouse model of atopic dermatitis. AB - Byakko-ka-ninjin-to (BN) is composed of gypsum, the root of anemarrhena, ginseng, licorice and rice. The effect of BN on the inhibition of itch was studied using an NC mouse model of atopic dermatitis. BN (200 mg/kg, p.o.) significantly inhibited the scratching frequency in NC mice, and decreased the skin temperature by 1.97 degrees C. The cooling action on the skin by BN may be involved in the inhibitory mechanism of itch, at least in part, since cooling the skin is known to inhibit the itch sensation in humans. Although the myocyte-specific enhancer binding factor 2C (MEF2C) mRNA is known to be increased in the cerebral cortex correlated with the itch sensation and skin lesions in NC mice, BN did not affect the expression level of the MEF2C mRNA. This result suggests that the inhibitory effect of BN on itch does not relate to inhibition of MEF2C expression in the cerebral cortex. The present study indicates that BN has an inhibitory effect on itch, and may be a useful antipruritic drug for atopic dermatitis. PMID- 10815014 TI - Myocardial protection against ischaemia-reperfusion injury by a Polygonum multiflorum extract supplemented 'Dang-Gui decoction for enriching blood', a compound formulation, ex vivo. AB - 'Dang-Gui Decoction for Enriching the Blood' (BE), a traditional Chinese formulation comprising Angelica sinensis and Astragalus membranaceus, is used for stimulating red blood cell production as well as enhancing cardiovascular function. In the present study, we have demonstrated the myocardial protection afforded by BE pretreatment against ischaemia-reperfusion (IR) injury in isolated perfused rat hearts. A more complete and potent myocardial protection against IR injury was also shown by a Polygonum multiflorum extract supplemented BE preparation (BEA). The results suggest that the more potent cardioprotective action of BEA may be related to its ability to sustain the myocardial glutathione antioxidant status under conditions of IR-induced oxidative stress, which may possibly in turn result from the synergistic interaction between the BE and Polygonum extract. PMID- 10815015 TI - A comparative study on hypocholesterolaemic effect of allicin, whole germinated seeds of bengal gram and guggulipid of gum gugglu. AB - The daily use of allicin and whole germinated bengal gram seeds for 8 weeks led to a significant decrease in serum cholesterol levels in normal volunteers with no side effects. The standard reference, guggulipid therapy, significantly (p < 0.001) reduced the mean serum cholesterol level to 142.88 +/- 24.14 mg/100 mL from an initial status of 211.91 +/- 6.23 mg/100mL (32.36% +/- 12.48% fall). Allicin treatment significantly reduced the mean serum cholesterol level to 129.99 +/- 2.79 mg/100 mL from a pre-treatment value of 148.10 +/- 8. 81 mg/100mL, a fall of 13.36% +/- 4.64%. The serum cholesterol value was reduced to 135.62+/- 22.85 mg/100 mL from a pre-treatment value of 157.29 +/- 4.94 mg/100mL (17.15% +/- 9.94% fall) due to daily use of whole germinated bengal gram seeds. Hence the hypocholesterolaemic effects of allicin and whole germinated bengal gram seeds are comparable to the established standard reference, guggulipid. These two materials are herbal in origin and it is supposed to have a lesser risk benefit ratio compared with other available synthetic drugs. The inclusion of these herbal products in the normal diet may be an alternative effective measure for hypercholesterolaemia. PMID- 10815016 TI - A study of the trypanocidal and analgesic properties from Lychnophora granmongolense (Duarte) Semir & Leitao Filho. AB - Crude extracts from the aerial parts of Lychnophora granmongolense (Asteraceae) were bioassayed for trypanocidal (trypomastigote forms of Trypanosoma cruzi) and analgesic (writhing test) activities. The crude ethyl acetate extract from the leaves plus inflorescences exhibited trypanocidal activity but no analgesic activity in the writhing model of pain. The bioassay-guided fractionation of this extract yielded three trypanocidal compounds: the sesquiterpene lactones centratherin (lychnophorolide A) and goyazensolide and the flavonoid eriodictyol. The flavonoids homoeriodictyol, eriodictyol 7, 3'-dimethyl ether, velutin, chrysoeriol, dihydroisorhamnetin, rhamnazin and the sesquiterpene lactone lychnophorolide B were also isolated from the ethyl acetate extract. Such flavonoids did not show any trypanocidal activity. The isolated amount of lychnophorolide B was not enough to account for the full activity against T. cruzi. PMID- 10815017 TI - Anticandidal activity of certain South Indian medicinal plants. AB - The anticandidal activity of 20 household South Indian medicinal plants and/or plant products was studied using 30 Candida albicans isolates obtained from vaginal candidiasis patients of Rajah Muthiah Medical College and Hospital and compared with the anticandidal activity of garlic. Water and ethanol extracts were prepared and their minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) were determined. Water extracts of three plants did not show any anticandidal activity, while Murrya koenigii did not exhibit any anticandidal activity in either extract. Other plants exhibited more activity in ethanol extracts showing that their active principle is more soluble in a non polar solvent. PMID- 10815018 TI - Screening for antimycotic properties of 56 traditional Chinese drugs. AB - In the present study we screened extracts of 56 widely used dried Chinese medical plants or their parts (TCD) for their antimycotic properties against pathological phyla of Aspergillus fumigatus, Candida albicans, Geotrichum candidum and Rhodotorula rubra. The highest activity against Aspergillus fumigatus was shown by Carthamus tinctorius L. (flos) and Rheum palmatum L. (radix et rhizoma) and against Candida albicans, Scutellaria baicalensis Georgi (radix) had the highest activity. The highest activity against Geotrichum candidum was shown by Agastache rugosa (Fisch et Mey.) O. Ktze. Herba Menthae has moderate antimycotic properties. PMID- 10815019 TI - Inhibition of enteric parasites by emulsified oil of oregano in vivo. AB - Oil of Mediterranean oregano Oreganum vulgare was orally administered to 14 adult patients whose stools tested positive for enteric parasites, Blastocystis hominis, Entamoeba hartmanni and Endolimax nana. After 6 weeks of supplementation with 600 mg emulsified oil of oregano daily, there was complete disappearance of Entamoeba hartmanni (four cases), Endolimax nana (one case), and Blastocystis hominis in eight cases. Also, Blastocystis hominis scores declined in three additional cases. Gastrointestinal symptoms improved in seven of the 11 patients who had tested positive for Blastocystis hominis. PMID- 10815020 TI - Protection by garlic against adriamycin induced alterations in the oxido reductive status of mouse red blood cells. AB - The effects of oral garlic supplementation on the activities of (a) the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPX) and (b) lipid peroxidation, as assessed by malondialdehyde (MDA) production in red blood cells of normal mice and those subject to oxidative stress by chronic administration of the anti-tumour drug adriamycin has been investigated. As expected, adria-mycin administration resulted in a significant increase in MDA generation (by 105.4%) and a decrease in GPX activity (by 23.8%) in the red blood cells. Although garlic had no significant effects on the basal levels of the antioxidant enzymes or MDA generation in red blood cells of normal mice (untreated with adriamycin), at doses of 20 mg/kg or 100 mg/kg, garlic was able to decrease significantly the adriamycin induced changes in the oxido-reductive status of the red blood cells. Thus, on administration of adriamycin to mice fed diets containing 20 mg/kg or 100 mg/kg garlic, the drug-induced increase in MDA generation was 38.2% and 22.5% respectively, less than that produced by adriamycin in mice fed normal diets, containing no garlic (105.4%). Similarly, in mice fed diets providing 20 mg/kg and 100 mg/kg garlic, adriamycin was able to decrease GPX activity by only 15.1% and 7.6% respectively, less than that produced by adriamycin in rats fed normal diets, containing no garlic (23.9%). PMID- 10815021 TI - Patents alert PMID- 10815022 TI - Selected bibliography PMID- 10815023 TI - S. Szolmicki, L. Samochowiec, J. Wojcicki and M. Drozdzik, 'The influence of active components of eleutherococcus senticosus on cellular defence and physical fitness in man'. Phytotherapy research 14(1) 2000, 30-35 AB - The original article to which this Erratum refers was published in Phytotherapy Research 14(1) 2000,30-35. Following the publication of this paper, it came to our attention that the lead author's name was published incorrectly. The correct name of the first author of the above paper is Jaroslaw Szolomicki. PMID- 10815024 TI - Is virology cost-effective. PMID- 10815025 TI - HSV-2 specific serology should be offered routinely to antenatal patients. AB - The most devastating consequence of genital herpes is neonatal herpes. It is clear that the majority of newborns acquire their infection by contact with infected genital secretions during delivery from an asymptomatic mother who acquired a first episode of genital herpes near the time of labour. Since the majority of cases of first episode genital herpes during pregnancy are unrecognised, the prevention of neonatal transmission will depend upon the identification of the HSV serologically discordant couple and the institution of appropriate interventions by mid pregnancy. Therefore, the precis of this discussion paper is that universal HSV serological testing should be performed at the first prenatal visit. As a corollary, type specific HSV serology will need to be commercially available and relatively inexpensive. In any country, pregnant women and their partners represent a broad, cross section of sexually active adults. The vast majority present themselves to the health care system for care during their pregnancies which is a period of time in which the focus of care is primarily preventive and during which women are generally motivated and compliant. This is truly the 'golden opportunity' to identify patients already infected as well as those at risk for acquiring genital herpes. Information regarding genital herpes and methods of preventing transmission to susceptible partners and newborn infants can easily be added to educational programmes which have already become an institution within prenatal care. PMID- 10815026 TI - HSV-2 specific serology should not be offered routinely to antenatal patients. AB - The proposal to introduce antenatal screening for HSV has no evidence for a public or individual health benefit; indeed, it has the potential to increase anxiety for patients, with a minimal likelihood of reducing the risk of neonatal herpes infection. Antenatal screening of an essentially healthy population of women must be validated in settings of different rates of neonatal HSV infection and the purposes and limitations of screening clearly outlined. Identification of pregnant women at risk of acquiring genital herpes in pregnancy is also dependent upon being able to obtain the serostatus of the male sexual partner which will reduce the practical application of the test if both patient and partners need to be screened. We recommend that efforts to improve on the currently established mechanisms for reducing the morbidity of neonatal herpes, namely early diagnosis and prompt treatment, must take priority for resources over new and unevaluated screening programmes, such as routine testing of antenatal patients. PMID- 10815027 TI - Human herpesvirus 6. AB - Human herpesvirus 6 (HHV-6), a member of the beta-herpesvirinae subfamily, is highly seroprevalent, has a worldwide distribution, and infection usually occurs within the first two years of life. In this age group, HHV-6 causes febrile illness including exanthem subitum with seizures a recognised complication. The virus is predominantly T lymphotropic although it can infect a variety of cell types in vitro and CD46 has recently been identified as a cellular receptor. The virus persists in the host, with a latent state proposed in monocytes and bone marrow progenitor cells, and chronic infection in salivary glands. The virus is pathogenic in the post transplantation period and may be a cofactor in the progression of HIV disease. The virus has also been associated with multiple sclerosis (MS), with the virus detected in oligodendrocytes particularly in plaque regions. The role of HHV-6 in MS remains controversial and a more extensive understanding of its neurotropism and association with disease is required. Two variants of HHV-6 exist (A and B) and comparison of their complete nucleotide sequences shows the genomes to be colinear, with a high degree of homology. Variation in specific regions of the genome is more extensive and probably accounts for biological and pathological differences. Almost exclusively, variant B is associated with febrile illness in childhood and is the predominant variant detected in healthy individuals. The epidemiology of HHV-6A infection needs to be better defined, although it is significantly less prevalent. Biological, genetic, epidemiological and pathological findings suggest that the two variants are divergent. PMID- 10815028 TI - Viral cyclins. AB - Cyclins are regulatory subunits of the cyclin-dependent protein kinases (CDKs). Members of this serine-threonine kinase family regulate the progression of cells through the division cycle. Until some years ago, cyclins were presumed to be encoded exclusively by eukaryotic cells. However, sequencing in 1996 of a simian herpesvirus, the herpesvirus saimiri, uncovered an open reading frame with sequence similarity to cellular cyclins. What at the time was a surprise for virologists and cell biologists alike, has become an accepted occurrence now. Eight different cyclin-encoding viruses have been described to date. One of them is the recently discovered human herpesvirus 8 (KSHV) suspected to cause Kaposi's sarcoma and certain B cell-lymphoproliferations in man. The significance of virus encoded cyclins in the viral life cycle is currently unclear. However, the link between specific cellular cyclins and cancer suggests that virus-encoded cyclins could be involved in oncogenic events associated with these cyclin-encoding viruses. PMID- 10815029 TI - Progress with retroviral gene vectors. AB - Retroviral vectors have become a standard tool for gene transfer technology. Compared with other gene transfer systems, retroviral vectors have several advantages, including their ability to transduce a variety of cell types, to integrate efficiently into the genomic DNA of the recipient cells and to express the transduced gene at high levels. The relatively well understood biology of retroviruses has made possible the development of packaging cell lines which provide in trans all the viral proteins required for viral particle formation. The design of different types of packaging cells has evolved to reduce the possibility of helper virus production. The host range of retroviruses has been expanded by pseudotyping the vectors with heterologous viral glycoproteins and receptor-specific ligands. The development of lentivirus vectors has allowed efficient gene transfer to quiescent cells. This review describes different strategies adopted for developing vectors to be used in gene therapy applications. PMID- 10815030 TI - Functional operation of the balance system in daily activities. AB - This article presents the anatomy and physiology of the balance system focusing on the daily use of the system in normal individuals to accomplish the major goals of the system. These goals include perceptions of movement and orientation, control of eye movements for clear visualization of the world, and control of upright stance during static and dynamic tasks. When discussing various pathologies and the assessment and treatment options available, the information provided sets the foundation on which the remainder of the articles are built. PMID- 10815031 TI - Neuro-otologic history. AB - Throughout medicine, the clinical history is the most important diagnostic tool. This is particularly true in vestibular disease, where pathologic confirmation of the disease process is rare. Many vestibular conditions are more appropriately called syndromes, rather than diseases, because the pathology is either variable or unknown. Knowledge of the anatomy and physiology provides the basis of understanding the control of balance and the symptoms that might occur should something go wrong. History taking should cover the elements of the balance system, including vestibular function, vision, hearing, somatosensation, and motor function. PMID- 10815032 TI - Management options for acute versus chronic vertigo. AB - Optimal management of the dizzy patient depends on an accurate diagnosis and knowledge of medical, surgical, and rehabilitative options. Acute vertigo may represent labyrinthine dysfunction treated with medication or a more serious central nervous system process requiring prompt intervention to avoid long-term disability. Chronic vertigo, however, may require surgical or rehabilitative measures for lasting relief. This article outlines the rationale for treating both groups with a variety of therapeutic options. PMID- 10815033 TI - Bedside vestibular examination. AB - A careful neuro-otologic examination is important in the diagnosis of vestibular disorders. This article reviews the bedside examination, beginning with the underlying physiologic principles. Techniques for testing static and dynamic vestibulo-ocular and vestibulospinal function are summarized. Finally, the use of specific provocative maneuvers is described. PMID- 10815034 TI - Balance function testing: a rational approach. AB - Recent advances in the area of balance function testing have left clinicians with a wide variety of diagnostic tests that provide insights into the function of the balance system. Laboratory tests used to evaluate the balance system include electronystagmography, rotational chair, and dynamic posturography. This article summarizes the currently available balance function testing protocols. A cost effective and rational protocol for the application of these tests is provided. PMID- 10815035 TI - Outcome measures for assessment of treatment of the dizzy and balance disorder patient. AB - There are a variety of measurement tools available for assessing the treatment outcomes for patients with dizziness and imbalance. Some of these tools, however, may not be appropriate or valid for the patients in question. In this article, the various outcome measures are described and evaluated in terms of their reliability, validity, and sensitivity. There is no clearly superior outcome measure at this time, and the choice of the measurement tool depends on the patient and the aims of the treatment. PMID- 10815036 TI - Vertigo and dysequilibrium with associated hearing loss. AB - There are numerous disorders that can present with hearing loss and vertigo or dysequilibrium. The combination of vertigo and imbalance associated with hearing loss are symptoms suggestive of a peripheral vestibular disorder. This article summarizes presentation, diagnosis, and treatment of the various common and rare peripheral vestibular disorders that can present with these symptoms. PMID- 10815037 TI - Diagnosis and initiating treatment for peripheral system disorders: imbalance and dizziness with normal hearing. AB - Disorders affecting the peripheral vestibular system commonly involve the peripheral auditory system causing hearing loss. There are a number of disorders, however, that selectively involve the peripheral vestibular system causing dizziness without hearing loss. These disorders include benign paroxysmal positional vertigo, vestibular neuritis, recurrent vestibulopathy, familial vestibulopathy, and bilateral idiopathic vestibulopathy. This article reviews these disorders and their diagnosis and management. PMID- 10815038 TI - Distinguishing and treating causes of central vertigo. AB - A patient's dizziness can be caused by a peripheral vestibular disorder, VIIIth nerve compression, brain stem ischemia, or cerebellar stroke. Clues from the history and physical examination are mentioned, and diagnostic entities, such as demyelination, cerebrovascular disease, migraine, Arnold-Chiari malformation, cerebellar degeneration, and neoplastic disease are discussed. Treatment options are outlined so that therapeutic and diagnostic trials can be initiated. Guidelines are offered for when to image the brain or posterior circulation vasculature and when a patient with acute vertigo should be admitted for observation. PMID- 10815039 TI - Overview of psychologic effects of chronic dizziness and balance disorders. AB - Symptoms of dizziness and imbalance often appear to be inexplicable, unpredictable, and uncontrollable. Chronic dizziness frequently leads to psychologic problems, such as panic and anxiety, preoccupation with health, avoidance of activities that seem to cause dizziness, and avoidance of situations in which dizziness might result in physical harm or social embarrassment. Paradoxically, anxiety and avoidance of activity can exacerbate and perpetuate dizziness, resulting in a vicious cycle of persistent disorientation and distress. Excessive anxiety and handicap can be pre- vented by providing patients (and their families) with a simple but detailed explanation of how symptoms are provoked and the process of adaptation following a balance disorder. This information enables patients to understand, anticipate, and manage their own symptoms. PMID- 10815040 TI - Diagnosis and treatment of psychologic symptoms and psychiatric disorders in patients with dizziness and imbalance. AB - Patients with dizziness and psychopathology present diagnostic and therapeutic challenges to clinicians in several specialties. Clinical research is beginning to define the medical-psychiatric interactions that produce many of these patients' symptoms. Treatment research is still in its infancy. A model of illness progression is presented to guide physicians through the evaluation of this patient population. Treatment recommendations are based on current clinical experience. PMID- 10815041 TI - Assessment of unexplained falls and gait unsteadiness: the impact of age. AB - When a patient with a balance disorder reports rotational vertigo, the clinician rightly focuses his or her attention on the vestibular system. This article reviews the possible diagnoses in the many patients who primarily report falls or gait disorder. Falls can be caused by predisposing neurologic conditions impairing gait, cardiovascular conditions, or epileptic episodes. The proportion of idiopathic falls, however, remains high. In the elderly, environmental circumstances, visual defects, psychotropic medication, and poor general health are additional risk factors. Clinical assessment of gait is more revealing and less expensive than computerized posture/ gait systems. The diagnosis of orthostatic tremor, however, requires either Fourier analysis of sway platform signals or electromyography. PMID- 10815042 TI - Efficacy of vestibular rehabilitation. AB - This article is a summary of the research on the effectiveness of vestibular rehabilitation in patients with vestibular disorders. The literature on patients with peripheral, central, combined peripheral and central disorders, and panic and anxiety disorders is reviewed. The positive outcomes of intervention are highlighted. PMID- 10815043 TI - The terminology of skin disorders. AB - Accurate diagnosis of skin disorders is essential for primary care physicians; studies have shown that dermatologic diagnosis by primary care trainees and attending physicians could be improved. Careful attention to the history and a systematic approach to the physical examination are essential to our diagnostic skills. Thorough description of the history and physical findings are valuable for reference as the condition evolves and for communication when referral to a dermatologist is needed. This article reviews the lexicon of skin disorders, beginning with description of the component of skin affected, the primary lesion, the extent and distribution, the characteristics of the lesions, and secondary or special lesions. Careful description of skin color, moisture, turgor, texture, and temperature are emphasized, and definition of skin lesions follows. PMID- 10815044 TI - Pearls in the management of acne: an advanced approach. AB - Acne is a common condition of the sebaceous follicle. The primary care physician can have a large impact on patients with acne by properly classifying the type of acne (obstructive versus inflammatory) and successfully treating the acne based on its severity. Reduction of acne lesions by appropriate topical and oral medications provides great psychological and physical benefits to these patients. By understanding how acne develops and its many manifestations and treatment options, the primary care physician can become an expert in acne diagnosis and treatment. PMID- 10815045 TI - Rosacea. AB - Rosacea is a common skin disorder most often seen in individuals between the ages of 30 and 60. The condition frequently produces erythema, papules, pustules, and edema of midfacial skin. Ocular rosacea occurs in a high percentage of patients, and is a major cause of red eye. A variety of treatments exist that can eliminate pustules, but no therapy is highly effective in eliminating the vascular flushing associated with rosacea. PMID- 10815046 TI - Hair disorders. AB - Disorders of the hair are commonplace in the primary care practice. Among these disorders are male pattern baldness, Telogen effluvium, alopecia areata, Trichotillomania, and fungal infections involving the hair shaft. A review of the normal anatomy and life cycle of hair also is presented. PMID- 10815047 TI - Nail disorders. AB - Nail lesions can be related to a number of disease states. These may be intrinsic to the nail, caused by infection, or reflect systemic disease. Careful history and examination often is necessary for accurate diagnosis. Anatomy and gross changes of the nail are reviewed. The diagnosis and treatment of disease process such as psoriasis, lichen planus, paronychia, onychomycosis, myxoid cysts, and pincer nails are reviewed. PMID- 10815048 TI - Pediatric exanthems. AB - Many childhood diseases have cutaneous manifestations such as exanthems. More than 50 agents (viral, bacterial, or rickettsial) that cause exanthems in children have been identified. This article describes a systematic approach for evaluating 12 common exanthems. Diagnosis is made by correlating the location, morphology, and timing of the eruption with associated constitutional signs and symptoms. PMID- 10815049 TI - Cutaneous manifestations of diabetes mellitus. AB - A broad spectrum of cutaneous disorders may be encountered in patients with both type 1 and type 2 diabetes mellitus. On occasion, these dermatologic findings may even precede any clinical or biochemical evidence for diabetes. Cutaneous manifestations of diabetes mellitus can be classified as conditions with strong associations with diabetes mellitus; infectious causes of skin lesions; dermatologic disorders related to complications of diabetes mellitus; and skin conditions related to the treatment of diabetes mellitus. This article provides a concise review of the epidemiology, clinical findings, histopathologic features, differential diagnostic considerations, and therapeutic approaches to specific dermatologic disorders seen in diabetic patients. This article also provides a framework for an effective approach to the majority of cutaneous manifestations encountered in patients with diabetes mellitus in a typical primary care setting. PMID- 10815050 TI - Psoriasis. AB - Psoriasis is a common affliction that family practitioners and other primary care physicians are certain to encounter in their daily practice. This disease presents in many manifestations and in a diffuse patient population. The therapeutic tools available to combat this disease are numerous, as are their untoward effects. This article, directed at the primary care physician, is a review of the spectrum of psoriasis and its manifestations, diagnosis, and management. PMID- 10815051 TI - Fungal skin disorders. AB - Fungal skin infections are becoming more common, and primary care physicians must be familiar with the diagnosis and treatment of these disorders. Dermatophyte infections including tinea pedis, corporis and cruris; pityriasis versicolor; and cutaneous candidiasis are the focus of this article. Common presentations of these disorders, the differential diagnosis, and current treatments are stressed. With the newer agents, shorter treatment courses can be used, and oral therapy for widespread, resistant, or recurrent infections is discussed. PMID- 10815052 TI - Cutaneous warts: diagnosis and treatment. AB - Cutaneous warts are a common malady that affects all ages, but are most prevalent in children and young adults. Warts result from infection with human papillomavirus and can lead to a condition that may be painful, disfiguring, and recurrent, despite treatment. A multitude of therapeutic modalities exist, though no single agent is invariably effective. Successful treatment must take into consideration the patient's level of motivation, expense, and physician experience. PMID- 10815053 TI - Sun-induced skin changes. AB - Chronic sun exposure is responsible for changes in skin texture, pigmentation, vascularity, maturation, and production of neoplasms. A 78% decrease in the lifetime incidence of basal and squamous cell carcinoma has been reported with regular use of SPF 15 sunscreen for the first 18 years of life. Actinic keratoses, sun-induced premalignant lesions of the skin, can be eliminated by mechanical and medication therapies. 5-fluorouracil therapy can be effective treatment for widespread actinic damage. PMID- 10815054 TI - Basal and squamous cell carcinoma. AB - Nonmelanotic skin cancer, primarily basal and squamous cell carcinoma, represents a major health concern in both the United States and the world. Primary care physicians must be able to recognize these cancers and perform the appropriate diagnostic tests to confirm their clinical suspicion. Several biopsy techniques are available for the physician to use in their office, and once diagnosis is confirmed, a variety of treatment modalities exist that provide excellent cure rates. Nevertheless, primary prevention with the use of sunscreen, protective clothing, and wide-brimmed hats is necessary to reduce the incidence of this costly disease. PMID- 10815055 TI - Bacterial infections of the skin. AB - This article discusses common bacterial skin infections, including impetigo, cellulitis and erysipelas folliculitis, staphylococcal scalded skin syndrome, blistering distal dactylitis, furuncles and carbuncles, and pseudomonal infections such as external otitis and malignant external otitis, ecthyma gangrenosum, pseudomonal folliculitis, toe web infection, and erysipeloid. PMID- 10815056 TI - Management of skin trauma. AB - The majority of skin trauma incidents cause minor injury that can be treated effectively in the outpatient setting. A new approach to the closure of skin lacerations using tissue adhesive is presented and the current management of abrasions, blisters, burns, skin tears, and subungual hematomas are reviewed. An overview of wound dressings is provided to assist in obtaining the optimal wound healing environment. PMID- 10815057 TI - Rhus (Toxicodendron) dermatitis. AB - This article reviews the current fund of knowledge on poison ivy, poison oak, and poison sumac dermatitis. It is intended as a global summary to provide the primary care provider with the required information and sources for more esoteric academic pursuits. Toxicodendron characteristics, morphology, and biology are reviewed. The overall medical impact is delineated as well as the clinical manifestations, pathophysiology, prevention, and treatment. Historical perspectives are mentioned throughout, as are future trends in research. PMID- 10815058 TI - Atopic dermatitis. AB - Atopic dermatitis is a chronic skin disorder. It affects up to 15% of the childhood population in the United States and more than half of these patients into adulthood. A clinical diagnosis is made based on the superficial, inflammatory, erythematous and puritic eruptions. Control of symptoms involves patient and parent education with regard to skin care and avoidance of triggers. Treatment includes corticosteroids, with newer strategies emerging. PMID- 10815059 TI - A piece of my mind. Naming names. PMID- 10815060 TI - Teaching medical students in the ambulatory setting: strategies for success. PMID- 10815061 TI - JAMA 100 Years Ago: INSTRUMENT FOR USE IN PHOTOTHERAPY. PMID- 10815062 TI - International group of health experts issues global "declaration to stop TB". PMID- 10815063 TI - New and emerging dermatologic therapies presented at conference. PMID- 10815065 TI - The World in Medicine: Noninvasive Ventilation. PMID- 10815064 TI - The World in Medicine: Safe Blood in the Americas. PMID- 10815067 TI - The World in Medicine: Parlez Francais. PMID- 10815066 TI - The World in Medicine: Promising Alzheimer Drug. PMID- 10815068 TI - Protection for Human Subjects in Medical Research. PMID- 10815069 TI - Protection for Human Subjects in Medical Research. PMID- 10815070 TI - Protection for Human Subjects in Medical Research. PMID- 10815071 TI - Protection for Human Subjects in Medical Research. PMID- 10815072 TI - Protection for human subjects in medical research. PMID- 10815074 TI - How Should Physicians Involve Patients in Medical Decisions? PMID- 10815073 TI - Protection for human subjects in medical research. PMID- 10815075 TI - How Should Physicians Involve Patients in Medical Decisions? PMID- 10815076 TI - How should physicians involve patients in medical decisions? PMID- 10815077 TI - How should physicians involve patients in medical decisions? PMID- 10815078 TI - How should physicians involve patients in medical decisions? PMID- 10815080 TI - Diagnosis and Treatment of Traumatic Brain Injury. PMID- 10815079 TI - Diagnosis and treatment of traumatic brain injury. PMID- 10815081 TI - A prescription drug packaged in China and sold as an ethnic remedy. PMID- 10815082 TI - Intravenous ancrod for treatment of acute ischemic stroke: the STAT study: a randomized controlled trial. Stroke Treatment with Ancrod Trial. AB - CONTEXT: Approved treatment options for acute ischemic stroke in the United States and Canada are limited at present to intravenous tissue-type plasminogen activator, but bleeding complications, including intracranial hemorrhage, are a recognized complication. OBJECTIVE: To evaluate the efficacy and safety of the defibrinogenating agent ancrod in patients with acute ischemic stroke. DESIGN: The Stroke Treatment with Ancrod Trial (STAT), a randomized, parallel-group, double-blind, placebo-controlled trial conducted between August 1993 and January 1998. SETTING: Forty-eight centers, primarily community hospitals, in the United States and Canada. PATIENTS: A total of 500 patients with an acute or progressing ischemic neurological deficit were enrolled and included in the intent-to-treat analysis. INTERVENTIONS: Patients were randomly assigned to receive ancrod (n=248) or placebo (n =252) as a continuous 72-hour intravenous infusion beginning within 3 hours of stroke onset, followed by infusions lasting approximately 1 hour at 96 and 120 hours. The ancrod regimen was designed to decrease plasma fibrinogen levels to 1.18 to 2.03 micromol/L. MAIN OUTCOME MEASURES: The primary efficacy end point was functional status, with favorable functional status defined as survival to day 90 with a Barthel Index of 95 or more or at least the prestroke value, compared by treatment group. Primary safety variables included symptomatic intracranial hemorrhage and mortality. RESULTS: Favorable functional status was achieved by more patients in the ancrod group (42.2%) than in the placebo group (34.4%; P=.04) by the prespecified covariate adjusted analysis. Mortality was not different between treatment groups (at 90 days, 25.4% for the ancrod group and 23% for the placebo group; P=.62), and the proportion of severely disabled patients was less in the ancrod group than in the placebo group (11.8% vs 19.8%; P=.01). The favorable functional status observed with ancrod vs placebo was consistent in all subgroups defined for age, stroke severity, sex, prestroke disability, and time to treatment (< or = 3 or > 3 hours after stroke onset). There was a trend toward more symptomatic intracranial hemorrhages in the ancrod group vs placebo (5.2% vs 2.0%; P=.06), as well as a significant increase in asymptomatic intracranial hemorrhages (19.0% vs 10.7%; P=.01). CONCLUSION: In this study, ancrod had a favorable benefit-risk profile for patients with acute ischemic stroke. PMID- 10815083 TI - Serum uric acid and cardiovascular mortality the NHANES I epidemiologic follow-up study, 1971-1992. National Health and Nutrition Examination Survey. AB - CONTEXT: Although many epidemiological studies have suggested that increased serum uric acid levels are a risk factor for cardiovascular mortality, this relationship remains uncertain. OBJECTIVE: To determine the association of serum uric acid levels with cardiovascular mortality. DESIGN AND SETTING: Cross sectional population-based study of epidemiological follow-up data from the First National Health and Nutrition Examination Survey (NHANES I) from 1971-1975 (baseline) and data from NHANES I Epidemiologic Follow-up Study (NHEFS). PARTICIPANTS: A total of 5926 subjects who were aged 25 to 74 years and had serum uric acid level measurements at baseline. MAIN OUTCOME MEASURES: Ischemic heart disease mortality, total cardiovascular mortality, and all-cause mortality, compared by quartiles of serum uric acid level. RESULTS: In an average of 16.4 years of follow-up, 1593 deaths occurred, of which 731 (45.9%) were ascribed to cardiovascular disease. Increased serum uric acid levels had a positive relationship to cardiovascular mortality in men and women and in black and white persons. Deaths due to ischemic heart disease in both men and women increased when serum uric acid levels were in the highest quartile compared with the lowest quartile (men, >416 vs <321 micromol/L; risk ratio, 1.77 [95% confidence interval [CI], 1.08-3.98]; women, >333 vs <238 micromol/l; risk ratio, 3.00 [95% CI, 1.45 6.28]). Cox regression analysis showed that for each 59.48-micromol/L increase in uric acid level, cardiovascular mortality and ischemic heart disease mortality increased. Hazard ratios for men were 1.09 (95% CI, 1.02-1.18) and 1.17 (95% CI, 1.06-1.28), and for women were 1.26 (95% CI, 1.16-1.36) and 1.30 (95% CI, 1.17 1.45), respectively, after adjustment for age, race, body mass index, smoking status, alcohol consumption, cholesterol level, history of hypertension and diabetes, and diuretic use. Further analysis, stratifying by cardiovascular risk status, diuretic use, and menopausal status, confirmed a significant association of uric acid and cardiovascular mortality in all subgroups except among men using diuretics (n=79) and men with 1 or more cardiovascular risk factors (n=1140). CONCLUSION: Our data suggest that increased serum uric acid levels are independently and significantly associated with risk of cardiovascular mortality. PMID- 10815084 TI - Association between method of delivery and maternal rehospitalization. AB - CONTEXT: Despite nearly 4 million deliveries in the United States each year, minimal information exists on unintended health consequences following childbirth, particularly in relation to delivery method. OBJECTIVE: To assess the risk for maternal rehospitalization associated with cesarean or assisted vaginal delivery compared with spontaneous vaginal delivery. DESIGN: Retrospective cohort study of data from the Washington State Birth Events Record Database for 1987 through November 1, 1996. SETTING AND PARTICIPANTS: All primiparous women without selected chronic medical conditions who delivered live singleton infants in nonfederal short-stay hospitals in Washington State (N =256,795). MAIN OUTCOME MEASURES: Relative risks (RRs) of rehospitalization within 60 days of cesarean or assisted vaginal vs spontaneous vaginal deliveries. RESULTS: A total of 3149 women (1.2%) were rehospitalized within 60 days of delivery. In logistic regression analyses adjusting for maternal age, rehospitalization was found to be more likely among women with cesarean delivery (RR, 1.8; 95% confidence interval [CI], 1.6-1.9) or assisted vaginal delivery (RR, 1.3; 95% CI, 1.2-1.4) than among women with spontaneous vaginal delivery. Cesarean delivery was associated with significantly increased risks of rehospitalization for uterine infection, obstetrical surgical wound complications, and cardiopulmonary and thromboembolic conditions. Among women with assisted vaginal delivery, significant increased risks were seen for rehospitalization with postpartum hemorrhage, obstetrical surgical wound complications, and pelvic injury. CONCLUSIONS: Women with cesarean and assisted vaginal deliveries were at increased risk for rehospitalization, particularly with infectious morbidities. Effective strategies for preventing and controlling peripartum infection should be an obstetrical priority. PMID- 10815086 TI - The perilous state of academic medicine. PMID- 10815087 TI - Sustaining change in medical education. PMID- 10815085 TI - Antiretroviral drug resistance testing in adult HIV-1 infection: recommendations of an International AIDS Society-USA Panel. AB - OBJECTIVE: Assays for drug resistance testing in human immunodeficiency virus type 1 (HIV-1) infection are now available and clinical studies suggest that viral drug resistance is correlated with poor virologic response to new therapy. The International AIDS Society-USA sought to update prior recommendations to provide guidance for clinicians regarding indications for HIV-1 resistance testing. PARTICIPANTS: An International AIDS Society-USA 13-member physician panel with expertise in basic science, clinical research, and patient care involving HIV resistance to antiretroviral drugs was reconvened to provide recommendations for the clinical use of drug resistance testing. EVIDENCE AND CONSENSUS PROCESS: The full panel met regularly between January and October 1999. Resistance and resistance testing data appearing in the last decade through April 2000 and presentations at national and international research conferences were reviewed. Recommendations and considerations were developed by 100% group consensus, acknowledging that definitive data to support final recommendations are not yet available. CONCLUSIONS: Emerging data indicate that despite limitations, resistance testing should be incorporated into patient management in some settings. Resistance testing is recommended to help guide the choice of new regimens after treatment failure and for guiding therapy for pregnant women. It should be considered in treatment-naive patients with established infection, but cannot be firmly recommended in this setting. Testing also should be considered prior to initiating therapy in patients with acute HIV infection, although therapy should not be delayed pending the results. Expert interpretation is recommended given the complexity of results and assay limitations. PMID- 10815088 TI - The quest for the therapeutic organization. PMID- 10815089 TI - The evolving academic health center: strategies and priorities at Emory University. PMID- 10815090 TI - The academic health center: some policy reflections. PMID- 10815091 TI - The plight of academic health centers. PMID- 10815092 TI - Ancrod--is snake venom an antidote for stroke? PMID- 10815093 TI - HIV genotype and phenotype--arresting resistance? PMID- 10815094 TI - Payment for immunosuppression after organ transplantation. American Society of Transplantation. AB - Dramatic improvements in organ transplantation have meant that a growing number of patients must take expensive immunosuppressive medications for the rest of their lives. Currently, Medicare covers most transplantation procedures in the United States, but ends coverage for outpatient immunosuppressive medications after 36 months. Evidence suggests that at least some patients have reduced immunosuppression and their transplants fail because they cannot afford these medication costs. In the years since the advent of effective immunosuppressive therapy, the US Congress has struggled with this issue, and in 1999 temporarily extended medication coverage for eligible patients (based on age and disability) by 8 months. However, a more permanent solution is needed. We advocate that Medicare should cover the cost of all immunosuppressive therapy for all solid organ transplant recipients who cannot afford to pay. A number of potentially cost-effective approaches could be taken, but, in any case, something must be done to ensure that transplants do not fail because recipients cannot pay for immunosuppression. PMID- 10815095 TI - A piece of My mind: A simple observation. PMID- 10815096 TI - JAMA 100 years ago: the mad-stone PMID- 10815098 TI - Tissue engineering in the genitourinary system. PMID- 10815097 TI - A "safer" cigarette? Prove it, say critics. PMID- 10815099 TI - Africa needs more aid, new partnerships to fight malaria. PMID- 10815100 TI - Health agencies update: easy on the antioxidants PMID- 10815101 TI - Health agencies update: Variety=Destiny? PMID- 10815102 TI - Health agencies update: curbing teen steroid Use PMID- 10815103 TI - Health agencies update: An unkind Cut? PMID- 10815104 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815105 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815106 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815107 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815108 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815109 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815110 TI - Human papillomavirus testing as a screening tool for cervical cancer. PMID- 10815111 TI - Human papillomavirus testing as a screening tool for cervical cancer PMID- 10815112 TI - Elevated liver enzymes following initiation of antiretroviral therapy. PMID- 10815113 TI - Elevated liver enzymes following initiation of antiretroviral therapy PMID- 10815114 TI - Prognostic criteria for hospice eligibility. PMID- 10815115 TI - Prognostic criteria for hospice eligibility PMID- 10815116 TI - Cognitive-behavioral therapy, imipramine, or their combination for panic disorder: A randomized controlled trial. AB - CONTEXT: Panic disorder (PD) may be treated with drugs, psychosocial intervention, or both, but the relative and combined efficacies have not been evaluated in an unbiased fashion. OBJECTIVE: To evaluate whether drug and psychosocial therapies for PD are each more effective than placebo, whether one treatment is more effective than the other, and whether combined therapy is more effective than either therapy alone. DESIGN AND SETTING: Randomized, double blind, placebo-controlled clinical trial conducted in 4 anxiety research clinics from May 1991 to April 1998. PATIENTS: A total of 312 patients with PD were included in the analysis. INTERVENTIONS: Patients were randomly assigned to receive imipramine, up to 300 mg/d, only (n=83); cognitive-behavioral therapy (CBT) only (n=77); placebo only (n=24); CBT plus imipramine (n=65); or CBT plus placebo (n=63). Patients were treated weekly for 3 months (acute phase); responders were then seen monthly for 6 months (maintenance phase) and then followed up for 6 months after treatment discontinuation. MAIN OUTCOME MEASURES: Treatment response based on the Panic Disorder Severity Scale (PDSS) and the Clinical Global Impression Scale (CGI) by treatment group. RESULTS: Both imipramine and CBT were significantly superior to placebo for the acute treatment phase as assessed by the PDSS (response rates for the intent-to-treat [ITT] analysis, 45.8%, 48.7%, and 21.7%; P=.05 and P=.03, respectively), but were not significantly different for the CGI (48. 2%, 53.9%, and 37.5%, respectively). After 6 months of maintenance, imipramine and CBT were significantly more effective than placebo for both the PDSS (response rates, 37.8%, 39.5%, and 13.0%, respectively; P=.02 for both) and the CGI (37.8%, 42.1%, and 13.0%, respectively). Among responders, imipramine produced a response of higher quality. The acute response rate for the combined treatment was 60.3% for the PDSS and 64.1% for the CGI; neither was significantly different from the other groups. The 6-month maintenance response rate for combined therapy was 57.1% for the PDSS (P=.04 vs CBT alone and P=.03 vs imipramine alone) and 56.3% for the CGI (P=.03 vs imipramine alone), but not significantly better than CBT plus placebo in either analysis. Six months after treatment discontinuation, in the ITT analysis CGI response rates were 41.0% for CBT plus placebo, 31.9% for CBT alone, 19.7% for imipramine alone, 13% for placebo, and 26.3% for CBT combined with imipramine. CONCLUSIONS: Combining imipramine and CBT appeared to confer limited advantage acutely but more substantial advantage by the end of maintenance. Each treatment worked well immediately following treatment and during maintenance; CBT appeared durable in follow-up. JAMA. 2000;283:2529-2536 PMID- 10815117 TI - Standard short-course chemotherapy for drug-resistant tuberculosis: treatment outcomes in 6 countries. AB - CONTEXT: No large-scale study has investigated the impact of multidrug-resistant tuberculosis (TB) on the outcome of standard short-course chemotherapy under routine countrywide TB control program conditions in the World Health Organization's (WHO) directly observed treatment short-course strategy for TB control. OBJECTIVE: To assess the results of treatment with first-line drugs for patients enrolled in the WHO and the International Union Against Tuberculosis and Lung Disease's global project on drug-resistance surveillance. DESIGN AND SETTING: Retrospective cohort study of patients with TB in the Dominican Republic, Hong Kong Special Administrative Region (People's Republic of China), Italy, Ivanovo Oblast (Russian Federation), the Republic of Korea, and Peru. PATIENTS: New and retreatment TB cases who received short-course chemotherapy with isoniazid, rifampicin, pyrazinamide, and either ethambutol or streptomycin between 1994 and 1996. MAIN OUTCOME MEASURE: Treatment response according to WHO treatment outcome categories (cured; died; completed, defaulted, or failed treatment; or transferred). RESULTS: Of the 6402 culture-positive TB cases evaluated, 5526 (86%) were new cases and 876 (14%) were retreatment cases. A total of 1148 (20.8%) new cases and 390 (44.5%) retreatment cases were drug resistant, including 184 and 169 cases of multidrug-resistant TB, respectively. Of the new cases 4585 (83%) were treated successfully, 138 (2%) died, and 151 (3%) experienced short-course chemotherapy failure. Overall, treatment failure (relative risk [RR], 15.4; 95% confidence interval [CI], 10.6-22.4; P<.001) and mortality (RR, 3.73; 95% CI, 2.13-6.53; P<.001) were higher among new multidrug resistant TB cases than among new susceptible cases. Even in settings using 100% direct observation, cases with multidrug resistance had a significantly higher failure rate than those who were susceptible (9/94 [10%] vs 8/1410 [0.7%]; RR, 16.9; 95% CI, 6.6-42.7; P<.001). Treatment failure was also higher among patients with any rifampicin resistance (n=115) other than multidrug resistance (RR, 5.48; 95% CI, 3.04-9.87; P<.001), any isoniazid resistance (n=457) other than multidrug resistance (RR, 3. 06; 95% CI, 1.85-5.05; P<.001), and among patients with TB resistant to rifampicin only (n=76) (RR, 5.47; 95% CI, 2.68-11.2; P<.001). Of the retreatment cases, 497 (57%) were treated successfully, 51 (6%) died, and 124 (14%) failed short-course chemotherapy treatment. Failure rates among retreatment cases were higher in those with multidrug-resistant TB, with any isoniazid resistance other than multidrug resistance, and in cases with TB resistant to isoniazid only. CONCLUSIONS: These data suggest that standard short-course chemotherapy, based on first-line drugs, is an inadequate treatment for some patients with drug-resistant TB. Although the directly observed treatment short course strategy is the basis of good TB control, the strategy should be modified in some settings to identify drug-resistant cases sooner, and to make use of second-line drugs in appropriate treatment regimens. JAMA. 2000;283:2537-2545 PMID- 10815118 TI - Association of hostility with coronary artery calcification in young adults: the CARDIA study. Coronary Artery Risk Development in Young Adults. AB - CONTEXT: Psychosocial factors, including personality and character traits, may play a role in the development and expression of coronary artery disease. OBJECTIVE: To evaluate whether hostility, a previously reported predictor of clinical coronary artery disease, is associated with coronary calcification, which is a marker of subclinical atherosclerosis. DESIGN: Prospective cohort study. SETTING AND PARTICIPANTS: Volunteer subsample from Chicago, Ill, and Oakland, Calif, consisting of 374 white and black men and women, aged 18 to 30 years at baseline, who participated in the Coronary Artery Risk Development in Young Adults (CARDIA) study. Cook-Medley hostility assessment data were collected at baseline from 1985 to 1986 and at year 5 examinations from 1990 to 1992. After the 10-year examinations in the 1995-1996 year, electron-beam computed tomographic scans were performed. MAIN OUTCOME MEASURES: Presence of any detectable coronary artery calcification (coronary calcium score >0), and coronary artery calcium scores of 20 or higher. RESULTS: In logistic regression analysis adjusting for age, sex, race, and field center comparing those with hostility scores above and below the median of the distribution of the present sample, the odds ratio of having any coronary calcification was 2.57 (95% confidence interval, 1.31-5.22), and the odds ratio of having a calcium score of 20 or higher was 9.56 (95% confidence interval, 2.29-65.9) for calcium scores of 20 or higher. The associations with any coronary artery calcification persisted after adjusting for demographic, lifestyle, and physiological variables. Results using a cynical distrust subscale were somewhat weaker than for those using the global hostility score. Power was inadequate to perform sex- or race-specific analyses. CONCLUSION: These results suggest that a high hostility level may predispose young adults to coronary artery calcification. JAMA. 2000;283:2546 2551 PMID- 10815119 TI - Abnormal glucose metabolism and pancreatic cancer mortality. AB - CONTEXT: Previous studies reported an increased risk of pancreatic cancer among persons with diabetes. Few data exist, however, on the association of postload plasma glucose concentration with pancreatic cancer, which could provide insight into the role of abnormal glucose metabolism in the etiology of pancreatic cancer. OBJECTIVE: To determine the independent association between postload plasma glucose concentration and risk of pancreatic cancer mortality among persons without self-reported diabetes. DESIGN: Prospective cohort study. SETTING AND PARTICIPANTS: Employees of 84 Chicago-area organizations, with an average age of 40 years at baseline, were screened from 1963 to 1973 and followed up for an average of 25 years. A total of 96 men and 43 women died of pancreatic cancer among 20,475 men and 15,183 women, respectively. MAIN OUTCOME MEASURES: Relationship of pancreatic cancer mortality with postload plasma glucose levels. RESULTS: Compared with a postload plasma glucose level of 6.6 mmol/L (119 mg/dL) or less and after adjusting for age, race, cigarette smoking, and body mass index, the relative risks (95% confidence intervals) of pancreatic cancer mortality were 1.65 (1.05-2.60) for postload plasma glucose levels between 6.7 (120) and 8.8 (159) mmol/L (mg/dL); 1.60 (0.95-2.70) for levels between 8.9 (160) and 11.0 (199); and 2.15 (1.22-3.80) for levels of 11.1 (200) or more; P for trend=.01. An association appeared to be stronger for men than women. Estimates were only slightly lower after excluding 11 men and 2 women who died of pancreatic cancer during the first 5 years of follow-up. In men only, higher body mass index and serum uric acid concentration also were independently associated with an elevated risk of pancreatic cancer mortality. CONCLUSION: These results suggest that factors associated with abnormal glucose metabolism may play an important role in the etiology of pancreatic cancer. JAMA. 2000;283:2552-2558 PMID- 10815120 TI - Patient characteristics and ICU organizational factors that influence frequency of pulmonary artery catheterization. AB - CONTEXT: Hemodynamic monitoring of patients with a pulmonary artery catheter is controversial because there are few data confirming its effectiveness, and patient and intensive care unit (ICU) organizational factors associated with its use are unknown. OBJECTIVE: To determine pulmonary artery catheter use in relationship to type of ICU organization and staffing, and patient characteristics, including severity of illness and insurance coverage. DESIGN, SETTING, AND PATIENTS: Retrospective database study of 10,217 nonoperative patients who received treatment at 34 medical, mixed medical and surgical, and surgical ICUs at 27 hospitals during 1998 (patients were enrolled in Project IMPACT). MAIN OUTCOME MEASURES: Pulmonary artery catheter use based on severity of illness measured by the Simplified Acute Physiology Score, resuscitation status at ICU admission, and ICU organizational variables, including type, size, and model. RESULTS: A pulmonary artery catheter was used for 831 patients (8.1%) in the ICU. In multivariate analysis adjusted for severity of illness, age, diagnosis, and do-not-resuscitate status, full-time ICU physician staffing was associated with a two-thirds reduction in the probability of catheter use (odds ratio [OR], 0.36; 95% confidence interval [CI], 0.28-0.45). Higher catheter use was associated with white race (OR, 1.38; 95% CI, 1.10-1.72) and private insurance coverage (OR, 1.33; 95% CI, 1.10-1.60). Admission to a surgical ICU was associated with a 2-fold increase in probability of catheter use (OR, 2.17; 95% CI, 1.70-2.76) compared with either medical or mixed medical and surgical ICUs. CONCLUSION: Organizational characteristics of ICUs, insurance reimbursement, and race, as well as clinical variables, are associated with variation in practice patterns regarding pulmonary artery catheter use. Understanding such influences, combined with studies measuring clinical and economic outcomes, can contribute to the development of policies for the rational use of pulmonary artery catheters. JAMA. 2000;283:2559-2567 PMID- 10815121 TI - Pulmonary artery catheterization and clinical outcomes: National Heart, Lung, and Blood Institute and Food and Drug Administration Workshop Report. Consensus Statement. AB - OBJECTIVE: The efficacy and safety of the pulmonary artery catheter are under scrutiny because of its association with increased morbidity and mortality in observational studies. In response, the National Heart, Lung, and Blood Institute (NHLBI) and the US Food and Drug Administration (FDA) conducted the Pulmonary Artery Catheterization and Clinical Outcomes workshop in Alexandria, Va, on August 25 and 26, 1997, to develop recommendations regarding actions to improve pulmonary artery catheter utility and safety. PARTICIPANTS: The NHLBI and FDA planning task force selected a workshop chairperson, subcommittee chairs, and participants. Approximately 85 participants were selected for their collective expertise in critical care, pulmonary medicine, cardiovascular medicine and surgery, pediatrics, nursing, biostatistics, and medical economics. The meeting was open to industry representatives and other government and lay observers. This workshop was funded by the NHLBI and the FDA's Division of Devices. EVIDENCE: Published reports relating to the efficacy and safety of the pulmonary artery catheter, especially consensus documents developed by professional societies. CONSENSUS PROCESS: The planning task force disseminated materials, held teleconferences, and developed draft position papers prior to the workshop. These were modified during the workshop and thereafter in the course of several teleconferences, and presented to the entire group for final modifications and approval. CONCLUSIONS: A need exists for collaborative education of physicians and nurses in performing, obtaining, and interpreting information from the use of pulmonary artery catheters. This effort should be led by professional societies, in collaboration with federal agencies, with the purpose of developing and disseminating standardized educational programs. Areas given high priority for clinical trials were pulmonary artery catheter use in persistent/refractory congestive heart failure, acute respiratory distress syndrome, severe sepsis and septic shock, and low-risk coronary artery bypass graft surgery. JAMA. 2000;283:2568-2572 PMID- 10815122 TI - Panic disorder--it's real and it's treatable. PMID- 10815123 TI - The global problem of multidrug-resistant tuberculosis: the genie is out of the bottle. PMID- 10815124 TI - Use of the pulmonary artery catheter in critically ill patients: was invention the mother of necessity? PMID- 10815125 TI - Inequality in quality: addressing socioeconomic, racial, and ethnic disparities in health care. AB - Socioeconomic and racial/ethnic disparities in health care quality have been extensively documented. Recently, the elimination of disparities in health care has become the focus of a national initiative. Yet, there is little effort to monitor and address disparities in health care through organizational quality improvement. After reviewing literature on disparities in health care, we discuss the limitations in existing quality assessment for identifying and addressing these disparities. We propose 5 principles to address these disparities through modifications in quality performance measures: disparities represent a significant quality problem; current data collection efforts are inadequate to identify and address disparities; clinical performance measures should be stratified by race/ethnicity and socioeconomic position for public reporting; population-wide monitoring should incorporate adjustment for race/ethnicity and socioeconomic position; and strategies to adjust payment for race/ethnicity and socioeconomic position should be considered to reflect the known effects of both on morbidity. JAMA. 2000;283:2579-2584 PMID- 10815126 TI - The many faces of Charcot-Marie-Tooth disease. PMID- 10815127 TI - Neuropathy, heredity, and monoclonal gammopathy. PMID- 10815128 TI - Mild cognitive impairment or questionable dementia? PMID- 10815129 TI - Role of the low-density lipoprotein receptor-related protein in beta-amyloid metabolism and Alzheimer disease. AB - Deposition of beta-amyloid (A beta), a metabolite of approximately 4 kd of the amyloid precursor protein, is a critical pathological feature in Alzheimer disease. We postulate that deposition reflects an imbalance of A beta synthesis and clearance. Several pathways that impact A beta converge on a single receptor molecule, the low-density lipoprotein receptor-related protein (LRP). This multifunctional receptor is the major neuronal receptor both for apolipoprotein E (apoE, protein; APOE, gene) and for alpha2-macroglobulin (alpha2M, protein; A2M, gene), and it mediates clearance of apoE/A beta and alpha2M/A beta complexes. The LRP also interacts with the amyloid precursor protein itself. In this review, we highlight data that support a role for LRP in A beta metabolism and hypothesize that LRP therefore plays a critical role in Alzheimer disease. PMID- 10815130 TI - The molecular basis for understanding neurotrophins and their relevance to neurologic disease. PMID- 10815131 TI - National Institutes of Health consensus conference: tuberous sclerosis complex. PMID- 10815132 TI - Hereditary motor and sensory neuropathy type 2C is genetically distinct from types 2B and 2D. AB - BACKGROUND: Linkage analysis studies have identified 3 genetically different varieties of hereditary motor and sensory neuropathy type 2 (HMSN 2, also called Charcot-Marie-Tooth disease type 2, or CMT 2): HMSN 2A (linked to 1p35-p36), 2B (to 3q13-q22), and 2D (to 7p14). Hereditary motor and sensory neuropathy type 2C is characterized by diaphragmatic and vocal cord paresis; its disease locus has not been mapped. OBJECTIVE: To determine whether the HMSN 2C phenotype, previously shown not to be linked to the HMSN 2A locus, is linked to the HMSN 2B or HMSN 2D loci. DESIGN: Linkage analysis. SETTING AND PATIENTS: Thirty-three subjects, including 12 affected individuals and 11 individuals at risk, in a large family with HMSN 2C. RESULTS: Evidence was found against linkage of HMSN 2C phenotype to either the HMSN 2B or the 2D loci. CONCLUSIONS: HMSN 2C is genetically distinct from HMSN 2A, 2B, and 2D. We think that at least 4 genetically distinct varieties of autosomal dominant HMSN 2 exist. PMID- 10815133 TI - Predicting conversion to Alzheimer disease using standardized clinical information. AB - OBJECTIVE: To identify aspects of a standardized clinical assessment that can predict which individuals within the category of "questionable" Alzheimer disease (AD) have a high likelihood of converting to AD over time. DESIGN: Detailed semistructured interviews were performed at baseline and annually for 3 years. SETTING: University-based gerontology research program. PATIENTS: The patient population consisted of 165 individuals 65 years and older: 42 of the participants had a Clinical Dementia Rating (CDR) of normal cognition (CDR rating, 0.0) and 123 had a rating of questionable AD (CDR rating, 0.5). After 3 years of follow-up, 23 of the 123 subjects with questionable AD were diagnosed with probable AD. MAIN OUTCOME MEASURES: The interview was used to generate a summary measure based on the sum of 6 CDR categories, known as the Total Box Score. The responses to 32 selected questions from the interview also were examined. RESULTS: Likelihood of progression to AD during the follow-up period was strongly related to the Total Box Score. For example, more than 50% of individuals with a Total Box Score of 2.0 or higher at baseline developed AD during the follow-up interval, whereas about 10% of individuals with a Total Box Score of 1.0 or lower developed AD during this same period. Selected questions from the standardized clinical interview also were highly predictive of subsequent conversion to AD among the study population. Eight selected questions from the clinical interview at baseline, combined with the CDR Total Box Score, identified 88.6% of such individuals accurately (questionable group, 82/91; converter group, 19/23). CONCLUSIONS: A standardized clinical assessment can be used to identify the subgroup of individuals within the category of questionable AD who have a high likelihood of converting to AD over time. Subjects who met the criteria for questionable AD had a variety of trajectories during a 3-year follow up, suggesting that diverse factors may influence the functional changes observed in this population. PMID- 10815134 TI - Stereotaxic injection of IgG from patients with Alzheimer disease initiates injury of cholinergic neurons of the basal forebrain. AB - CONTEXT: The participation of an immune/inflammatory process in the pathomechanism of sporadic Alzheimer disease (AD) has been suggested by evidence for activated microglia and the potential therapeutic benefit of anti inflammatory medication. OBJECTIVE: To define a possible role for IgG in the immune/inflammatory process of AD in humans, we assayed the ability of IgG samples from patients with AD to target the injury to cholinergic neurons in rat basal forebrain in vivo. DESIGN: IgG purified from the serum or plasma from patients with AD and patients with other neurological disease who were used as control (DC) patients was injected stereotaxically into the medial septum of adult rats. Four weeks later coronal sections of the whole medial septum-diagonal bands of Broca region were immunostained for choline acetyltransferase (ChAT) to identify cholinergic neuronal cells. SETTING: University medical centers. PATIENTS: Blood samples were collected from 8 patients with probable and definite AD and from 6 age-matched DC patients. MAIN OUTCOME MEASURE: Detection of changes in the number of ChAT immunopositive cell profiles in sections and statistical evaluation. RESULTS: Four weeks after the injections, IgG samples from patients with AD significantly reduced the number of ChAT-immunostained cell profiles in the whole medial septum-diagonal bands of Broca region compared with IgGs from DC patients. Neither DC IgGs nor saline solution significantly decreased the number of ChAT-immunopositive neuronal cell profiles. CONCLUSION: Data document that IgG from patients with AD can target a stereotaxically induced immune/inflammatory injury to cholinergic neurons in the rat basal forebrain in vivo. PMID- 10815135 TI - Prediction of spinal epidural metastases. AB - CONTEXT: Early diagnosis and treatment of spinal epidural metastases (SEM) is of the utmost importance to prevent neurological deficit due to spinal cord compression. Magnetic resonance imaging (MRI) has become the final tool in that diagnostic process. However, access to MRI is still limited in The Netherlands, requiring cost-effective use. It is generally acknowledged that patients with systemic cancer who present with a radiculopathy or myelopathy should undergo an MRI. However, the diagnostic policy in patients with systemic cancer who present with recently developed back pain is still a matter of debate. OBJECTIVE: To identify the patients with back pain in whom MRI can safely be omitted because of a low risk of SEM. METHODS: In a prospective series of 170 consecutive patients with cancer with recently developed back pain, prediction of spinal metastatic disease (SMD) and especially SEM was studied by means of a multivariate risk analysis of the parameters of the standard neurological evaluation (medical history, neurological examination, and plain films of the whole spine). Magnetic resonance imaging was used as the criterion standard. We calculated the risk implications of omitting MRI in patients with an estimated risk below different cutoff points. RESULTS: Spinal metastatic disease was diagnosed in 80 patients (47%); of these, 31 had SEM. A metastatic abnormality on plain films was the strongest independent predictor for SMD. Other important predictors were night pain, progressive pain, and Karnofsky score. Advanced age, exacerbation of pain during recumbency, and osteoporotic fracture imply a low risk of SMD. Night pain and the Karnofsky score proved to be the main predictors for SEM. A plain film showing an osteoporotic fracture strongly decreased the risk of SEM. The discriminating value of the multivariate analysis was too low, and too few patients can be excluded from undergoing MRI on the basis of the standard neurological checkup. To identify all the patients with SMD (P<.01), MRI would be excluded in only 7 patients. Identification of all patients with SEM (P<.001) reduced the number of MRIs by 21 at the expense of plain films of the whole spine for any patient. CONCLUSIONS: Selection of patients with cancer with back pain at risk of SEM was not possible with the standard neurological checkup. After intake by the neurologist, the next step should be MRI of the whole spine. PMID- 10815136 TI - No association between a presenilin 1 polymorphism and Alzheimer disease. AB - BACKGROUND: Homozygosity of allele 1 of a presenilin 1 intron 8 polymorphism (PS1 1) has been associated with doubling of the risk of sporadic late-onset Alzheimer disease (LOAD), in some, but not all studies. OBJECTIVE: To genotype the PS1 intron 8 polymorphism in predominantly Hispanic families with LOAD to test for association and for linkage between this polymorphism and LOAD. DESIGN: A family based, case-control, genetic-linkage study. SETTING: Predominantly Hispanic families were selected from probands who were part of a random sample of 2128 Medicare beneficiaries aged 65 years or older who were residing in the community of Washington Heights, which is located in the northern part of Manhattan, NY. PARTICIPANTS: Fifty-one families with 103 affected family members, 67 unaffected family members, and 7 family members with other diagnoses were genotyped for the PS1 polymorphism. All patients met National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria for either probable or possible Alzheimer disease. Age was truncated at 55 years or older. MAIN OUTCOME MEASURES: Association analyses, conditional logistic regression, and traditional linkage methods were applied to the families for the PS1 polymorphism and for the presence of the gene for apolipoprotein E (APOE). Results of the association and conditional logistic regression analyses of PS1 intron 8 polymorphism were subsequently adjusted for the effect of APOE-epsilon4, sex, age, and education of each sibling. RESULTS: No association between the PS1 intron 8 polymorphism and LOAD was observed (relative risk, 0.99; 95% confidence interval, 0.3-3.4). An association between presence of the APOE-epsilon4 allele and LOAD (relative risk 4.05; 95% confidence interval, 1.3-12.5) was observed. CONCLUSION: We could not confirm the relationship between the PS1 intron 8 polymorphism and LOAD in this collection of families. PMID- 10815137 TI - Electroencephalographic, volumetric, and neuropsychological indicators of seizure focus lateralization in temporal lobe epilepsy. AB - CONTEXT: Anterior temporal lobectomy is an effective treatment for medically intractable temporal lobe seizures. Identification of seizure focus is essential to surgical success. OBJECTIVE: To examine the usefulness of presurgical electroencephalography (EEG), magnetic resonance imaging (MRI), and neuropsychological data in the lateralization of seizure focus. DESIGN: Presurgical EEG, MRI, and neuropsychological data were entered, independently and in combination, as indicators of seizure focus lateralization in discriminant function analyses, yielding correct seizure lateralization rates for each set of indicators. SETTING: Comprehensive Epilepsy Program, Shands Teaching Hospital, University of Florida, Gainesville. PATIENTS: Forty-four right-handed adult patients who ultimately underwent successful anterior temporal lobectomy. Left handed patients, those with less-than-optimal surgical outcome, and any patients with a history of neurological insult unrelated to seizure disorder were excluded from this study. MAIN OUTCOME MEASURES: For each patient presurgical EEG was represented as a seizure lateralization index reflecting the numbers of seizures originating in the left hemisphere, right hemisphere, and those unable to be lateralized. Magnetic resonance imaging data were represented as left-right difference in hippocampal volume. Neuropsychological data consisted of mean scores in each of 5 cognitive domains. RESULTS: The EEG was a better indicator of lateralization (89% correct) than MRI (86%), although not significantly. The EEG and MRI were significantly superior to neuropsychological data (66%) (P=.02 and .04, respectively). Combining EEG and MRI yielded a significantly higher lateralization rate (93%) than EEG alone (P<.01). Adding neuropsychological data improved this slightly (95%). CONCLUSIONS: The EEG and MRI were of high lateralization value, while neuropsychological data were of limited use in this regard. Combining EEG, MRI, and neuropsychological improved focus lateralization relative to using these data independently. PMID- 10815138 TI - The relative frequency of "dementia of unknown etiology" increases with age and is nearly 50% in nonagenarians. AB - CONTEXT: With the recent change in pathological criteria for Alzheimer disease (AD), a group of patients has emerged who do not meet pathological criteria for any well-characterized degenerative dementias. Whether these unclassified patients have vascular dementia or some other form of dementia is not known. OBJECTIVE: To determine the clinical characteristics, pathological substrate, and relative frequency of dementia not caused by well-characterized degenerative dementias. DESIGN/SETTING: Clinicopathological study of a prospectively observed sample of elderly nondemented and demented subjects recruited from our urban community. METHODS: In our series of 128 subjects with prospective neuropsychological evaluations as well as neuropathology, we identified 35 clinically nondemented subjects and 20 demented patients who did not meet pathological criteria for well-characterized degenerative dementias such as AD or dementia with Lewy bodies. The 20 demented patients were grouped together under the term dementia of unknown etiology (DUE). We compared clinical, genetic, neuropsychological, pathological, and neurochemical characteristics of the nondemented group, patients with DUE, and 28 patients with AD and no other pathological abnormality. RESULTS: Mean age at death for patients with DUE was 89.1 +/- 5.8 years compared with 79.9 +/- 11.4 years for AD (P<.001). Patients with AD and DUE did not differ in sex, risk factors, apolipoprotein E genotype, neuropsychological features, or neurological features. Hippocampal sclerosis (in 11 patients with dementia and no controls) and leukoencephalopathy (in 7 patients with dementia and 1 control) were associated with cognitive impairment; other vascular markers were not. Dementia of unknown etiology accounted for 5% of all cases of dementia among patients dying in their 70s, 21% for patients dying in their 80s, and 48% for patients dying in their 90s. CONCLUSIONS: A significant percentage of demented patients older than 80 years do not meet pathological criteria for AD or dementia with Lewy bodies. Hippocampal sclerosis and leukoencephalopathy are common in these patients but rare in clinically nondemented subjects. PMID- 10815139 TI - Ethnic differences in essential tremor. AB - BACKGROUND: Ethnic differences in the clinical characteristics (severity and distribution) of essential tumor (ET) have not been studied. The presence of these differences suggests that ET is not a homogeneous disease and that there is variability in disease expression under different circumstances. As part of a community-based study, we evaluated a multiethnic group of cases. OBJECTIVE: To assess whether there are ethnic differences in the clinical characteristics of ET. METHODS: Elderly residents of Washington Heights-Inwood, New York, were enrolled in a community-based health study (N = 2117). Participants underwent a medical interview and a neurological examination conducted by a neurologist, and subjects with ET were identified. These subjects with ET were then enrolled in a community-based study of ET and underwent a tremor interview, a videotaped tremor examination, and in some cases, a performance-based test of function and quantitative computerized tremor analysis. A total tremor score (range, 0-36, with 0 indicating no tremor and 36 indicating maximum tremor) was assigned to each subject based on 2 neurologists' ratings of the tremor examination. RESULTS: Among 62 subjects with ET (white [n = 16], African American [n = 18], and Hispanic [n = 28]), there were ethnic differences in the total tremor score (F = 3.68, P = .03). In a multiple regression model adjusting for age, white subjects had a mean total tremor score that was 5.3 points lower than that of nonwhite subjects (P = .008). We divided the nonwhite group into African American and Hispanic subgroups. In a regression model adjusting for age and duration, the white group had a mean total tremor score that was 6.1 points lower than that of the Hispanic group (P = .07) and 7.2 points lower than that of the African American group (P = .05). The mean performance-based test score was 1.7 times higher in the African American group and 2.1 times higher in the Hispanic group compared with the white group (P = .38). No subjects in the African American group had head tremor, while 4 subjects in the white group (25%) and 8 subjects in the Hispanic group (29%) did have head tremor (chi2 = 6.17, P = .05). CONCLUSIONS: There are ethnic differences in the expression of ET, suggesting that ET is not a homogeneous disorder. These differences may reflect phenotypic variability caused by genotypic differences or differences in exposure to environmental factors that influence tremor. PMID- 10815140 TI - Clinical characteristics of pramipexole-induced peripheral edema. AB - BACKGROUND: Pramipexole, a new dopamine agonist, effectively treats early and advanced Parkinson disease and restless legs syndrome. OBJECTIVES: To report the clinical features of and to investigate the predisposing factors and eventual outcomes in patients who developed peripheral edema (PE) following treatment with pramipexole. DESIGN: Retrospective case series in a tertiary referral center. RESULTS: Of the 300 patients who were receiving pramipexole therapy, 17 patients had mild to severe PE, which was attributable to the medication. Fifteen patients had Parkinson disease and 2 patients had restless legs syndrome. The mean (+/-SD) time of onset of PE after pramipexole therapy was started was 2.6 +/- 3.6 months (range, 0.25-11 months). The mean (+/-SD) dose at onset of PE was 1.7 +/- 1.0 mg/d (dose range, 0.75-3 mg/d) and the mean (+/-SD) dose when PE was at its maximum was 2.6 +/- 0.7 mg/d (dose range, 1.5-3 mg/d). In all cases, the PE rapidly abated with discontinuation of therapy, and in all cases that we rechallenged, it rapidly returned. The condition seemed to be dose dependent but also idiosyncratic, as we could not identify any predisposing features. It resulted in extensive medical evaluation in some patients and was only minimally responsive to diuretic therapy. CONCLUSION: Peripheral edema should be included among the potential adverse events associated with pramipexole therapy. PMID- 10815141 TI - Hereditary spastic paraplegia and hereditary ataxia, Part 2: A family demonstrating various phenotypic manifestations with the SCA3 genotype. AB - BACKGROUND: Clinical descriptions of the dominantly inherited ataxic motor syndromes in a 7-generation family of German origin were first reported in 1951. OBJECTIVE: To provide follow-up clinical, pathological, and genetic data for 9 patients in this family. DESIGN: Clinical histories and neurologic findings, gross and microscopic pathological features, and DNA analysis. RESULTS: Clinical presentations in this closely followed up portion of the family include fairly uniform ataxic and upper motor neuron symptoms. Nystagmus was a conspicuous and early sign, but generational anticipation was not evident. Although often present, amyotrophy was not a major source of disability. Major pathological degeneration was noted in the pons, spinal cord, and upper brainstem, where ubiquitin-immunoreactive intranuclear inclusion bodies were demonstrated. The diagnosis of Machado-Joseph disease (SCA3 [spinocerebellar ataxia type 3] genotype) was established from autopsy tissue in 1 patient and from blood specimens in 6 others. CONCLUSIONS: Clinical variation within this family and between this family and families with the SCA1 and SCA3 genotypes is so broad as to make the genetic diagnosis from clinical criteria alone practically impossible. The pathological definition of Machado-Joseph disease is more reliable, but some findings do overlap those of other genotypes. To our knowledge, the basis for the phenotypic variations in Machado-Joseph disease, genetic or otherwise, has not been established. PMID- 10815142 TI - Three families with polyneuropathy associated with monoclonal gammopathy. AB - OBJECTIVE: To report familial occurrence of polyneuropathy associated with monoclonal gammopathy. DESIGN: Case reports. PATIENTS: We describe 6 patients (3 pairs) with a polyneuropathy associated with IgM monoclonal gammopathy. Four of the 6 patients had a demyelinating polyneuropathy on neurophysiological examination. Three patients had elevated antibodies against myelin-associated glycoprotein. No duplication on chromosome 17 or a mutation on chromosome 1 was found in any family. CONCLUSION: Familial occurrence of polyneuropathy without the presence of hereditary motor and sensory neuropathy type I is a reason to search for the presence of monoclonal gammopathy. PMID- 10815143 TI - Idiopathic stabbing headache associated with monocular visual loss. AB - BACKGROUND: Idiopathic stabbing headache, which is a brief, sharp, severe jabbing pain that is confined to the head, responds well to treatment with indomethacin sodium. It may occur as a primary entity but more likely is associated with other types of headache, including migraine. SETTING: Emergency department of a teaching hospital. PATIENT: A 27-year-old man presented to the emergency department with stabbing, sharp pain in the right temporal area associated with complete of loss vision in his right eye. The patient had a history of migraine with aura since 1995. RESULT: The patient was treated with oxygen inhalation and indomethacin, with complete resolution of his symptoms. CONCLUSION: A case of idiopathic stabbing headache associated with monocular visual loss was relieved by oxygen and indomethacin therapy. PMID- 10815144 TI - Images in neurology. Functional magnetic resonance imaging localization of motor function. PMID- 10815145 TI - Treatment of ocular myasthenia with corticosteroids: yes. PMID- 10815146 TI - Treatment of ocular myasthenia: steroids only when compelled. PMID- 10815147 TI - Treatment of ocular myasthenia. PMID- 10815148 TI - Carpal tunnel syndrome. PMID- 10815149 TI - Levodopa-treated Parkinson disease has better long-term outcome than previously predicted. PMID- 10815151 TI - Chronic Cryptogenic Sensory Polyneuropathy. PMID- 10815150 TI - Chronic cryptogenic sensory polyneuropathy. PMID- 10815152 TI - Reliability and validity of the Ocular Surface Disease Index. AB - OBJECTIVE: To evaluate the validity and reliability of the Ocular Surface Disease Index (OSDI) questionnaire. METHODS: Participants (109 patients with dry eye and 30 normal controls) completed the OSDI, the National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25), the McMonnies Dry Eye Questionnaire, the Short Form-12 (SF-12) Health Status Questionnaire, and an ophthalmic examination including Schirmer tests, tear breakup time, and fluorescein and lissamine green staining. RESULTS: Factor analysis identified 3 subscales of the OSDI: vision related function, ocular symptoms, and environmental triggers. Reliability (measured by Cronbach alpha) ranged from good to excellent for the overall instrument and each subscale, and test-retest reliability was good to excellent. The OSDI was valid, effectively discriminating between normal, mild to moderate, and severe dry eye disease as defined by both physician's assessment and a composite disease severity score. The OSDI also correlated significantly with the McMonnies questionnaire, the National Eye Institute Visual Functioning Questionnaire, the physical component summary score of the Short Form-12, patient perception of symptoms, and artificial tear usage. CONCLUSIONS: The OSDI is a valid and reliable instrument for measuring the severity of dry eye disease, and it possesses the necessary psychometric properties to be used as an end point in clinical trials. PMID- 10815153 TI - Correlation of histologic 2-dimensional reconstruction and confocal scanning laser microscopic imaging of choroidal neovascularization in eyes with age related maculopathy. AB - OBJECTIVE: To topographically localize vascular channels, macrophages, and retinal pigment epithelium and other components of choroidal neovascularization (CNV) associated with age-related maculopathy. METHODS: Two postmortem eyes with age-related maculopathy and CNV were evaluated. The formalin-fixed CNV complex was excised and processed for confocal scanning laser microscopy including immunostaining for factor VIII-related antigen and incubation with Ig fluorescein isothiocyanate. After confocal microscopy, the specimens were serial step sectioned, stained, and 2-dimensional topographic reconstructions were made. The confocal images were compared with the 2-dimensional reconstructions. RESULTS: Both specimens contained central disciform scars surrounded by areas of intact retinal pigment epithelium. The first specimen was more atrophic and contained fewer choroidal neovascular channels than the second specimen. The topographic arrangement of the CNV and retinal pigment epithelial changes in the confocal images corresponded with the 2-dimensional reconstructions. Macrophages were concentrated around areas of vascularization. CONCLUSION: Confocal scanning laser microscopy of excised CNV simulates fluorescein angiography and topographic localization of the components of CNV provides insight into the pathogenesis of CNV. PMID- 10815154 TI - Indocyanine green angiography of well-defined plaque choroidal neovascularization in age-related macular degeneration. AB - OBJECTIVE: To determine the natural course of well-defined plaque choroidal neovascularization (CNV) using indocyanine green angiography. METHODS: Two ophthalmologists, experts in macular diseases and indocyanine green angiography, examined 40 eyes with exudative age-related macular degeneration and a well defined plaque CNV using complete ophthalmoscopic evaluation, fluorescein angiography, and indocyanine green angiography. The increase in the size of the plaques was analyzed using multivariate analysis, in relation to the worsening of visual acuity, with adjustment for age, sex, and length of follow-up. RESULTS: Mean follow-up was 13.5 months (median, 11 months). Initial and final mean visual acuity were 20/46 (median, 20/50) and 20/65 (median, 20/100), respectively. The mean initial size of the plaque was 6.62 mm2 (median, 6.20 mm2), and the mean final size was 10.40 mm2 (median, 9.76 mm2). The enlargement was statistically significant (P<.001). CONCLUSIONS: We found that plaque CNV tends to become larger with time, the enlargement reaching about 40% in 1 year of follow-up. The resulting loss of visual acuity, however, is not significant, and is slightly correlated with the extension of the lesion; it also does not appear to be directly related to sex. PMID- 10815155 TI - Acquired tumors arising from congenital hypertrophy of the retinal pigment epithelium. AB - BACKGROUND: Congenital hypertrophy of the retinal pigment epithelium (CHRPE) is widely recognized to be a flat, stationary condition. Although it can show minimal increase in diameter, it has not been known to spawn nodular tumor that is evident ophthalmoscopically. OBJECTIVES: To report 5 cases of CHRPE that gave rise to an elevated lesion and to describe the clinical features of these unusual nodules. METHODS: Retrospective medical record review. RESULTS: Of 5 patients with a nodular lesion arising from CHRPE, there were 4 women and 1 man, 4 whites and 1 black. Three patients were followed up for typical CHRPE for longer than 10 years before the tumor developed; 2 patients were recognized to have CHRPE and the elevated tumor concurrently. Visual acuity was decreased in 3 patients, mainly due to cystoid macular edema. The tumor was located between the equator and ora serrata in all 5 patients. There was no predilection for quadrant of the fundus. The flat part of the lesion was black and had visible lacunae in all 5 patients. The CHRPE ranged in basal diameter from 3 x 3 mm to 13 x 11 mm. The size of the elevated lesion ranged from 2 x 2 x 2 mm to 8 x 8 x 4 mm. The nodular component in all cases was supplied and drained by slightly prominent, nontortuous retinal blood vessels. Yellow retinal exudation occurred adjacent to the nodule in all 5 patients and 1 patient developed a secondary retinal detachment. Two tumors that showed progressive enlargement, increasing exudation, and progressive visual loss were treated with iodine 125-labeled plaque brachytherapy, resulting in deceased tumor size but no improvement in the visual acuity. CONCLUSIONS: Congenital hypertrophy of the retinal pigment epithelium can spawn a nodular growth that slowly enlarges, attains a retinal blood supply, and causes exudative retinopathy and chronic cystoid macular edema. Although no histopathologic evidence is yet available, we believe that the tumor probably represents either an acquired adenoma or a reactive proliferation of the retinal pigment epithelium. The best treatment of these lesions is not yet established. PMID- 10815156 TI - Involution of retinopathy of prematurity. Cryotherapy for Retinopathy of Prematurity Cooperative Group. AB - OBJECTIVE: To report the timing of involution of acute retinopathy of prematurity (ROP). DESIGN: An analysis of prospective retinal observational data recorded at infants' eye examinations. PARTICIPANTS: Infants from the Multicenter Trial of Cryotherapy for Retinopathy of Prematurity (CRYO-ROP) had birth weights less than 1251 g and served as subjects. The study population included 766 children who were examined in 5 of the 23 study centers and who developed at least 1 clock hour of acute ROP, stages 1 through 3. One eye from each patient was randomly chosen for analysis. MAIN OUTCOME MEASURES: Investigators documented the location, extent, and severity of ROP during serial retinal examinations. The onset of the ROP's involution was determined from a review of these data, applying a set of predetermined criteria. RESULTS: Acute-phase ROP began to involute at a mean of 38.6 weeks postmenstrual age. In 90% of patients, the ROP began to involute before 44 weeks of postmenstrual age. Acute ROP that demonstrated involution by moving from zone II to zone III was associated with an unfavorable outcome in 2 (1%) of 200 cases. Retinopathy of prematurity that was present only in zone III during a child's serial retinal examinations was never associated with the development of a partial or total retinal detachment. CONCLUSIONS: The onset of involution of acute retinopathy of prematurity correlates better with postmenstrual rather than with chronological age. This is reminiscent of the reported similar correlation of postmenstrual age to the time of onset of prethreshold and threshold ROP. Zone III ROP was nearly always associated with a favorable outcome. PMID- 10815157 TI - Selective photodynamic effects of the new photosensitizer ATX-S10(Na) on choroidal neovascularization in monkeys. AB - OBJECTIVE: To determine the optimal treatment variables for photodynamic therapy (PDT) with new photosensitizer ATX-S10(Na) (13,17-bis[1-carboxypropionyl] carbamoylethyl-8-etheny-2-hydroxy-3-hydroxyiminoethyliden e-2,7,12,18-tetranethyl 6 porphyrin sodium) to induce selective occlusion of choroidal neovascularization (CNV) in nonhuman primate eyes. METHODS: Experimental CNV was induced in monkey eyes by laser photocoagulation, and PDT was performed in neovascularized and healthy eyes with different treatment variables. At 0 to 150 minutes after 4-, 8 , and 12-mg/kg of body weight intravenous injections of ATX-S10(Na), a diode laser was irradiated at the dose of 1 to 127 J/cm2 (wavelength, 670 nm). Vascular occlusion induced by PDT was evaluated using fluorescein angiography, indocyanine green angiography, and histological examination at 1 day to 4 weeks after irradiation. RESULTS: Selective occlusion of CNV without damage to healthy retinal and choroidal capillaries was achieved in the following conditions: 30 to 74 J/cm2 irradiation at 30 to 74 minutes after the 8-mg/kg injection, and 1 to 29 J/cm2 irradiation at 30 to 74 minutes or 30 to 74 J/cm2 irradiation at 75 to 150 minutes after the 12-mg/kg dye injection. Regrowth of CNV often occurred when the retina was heavily injured by excessive PDT. CONCLUSION: By using optimal treatment variables, PDT using ATX-S10(Na) induces selective occlusion of CNV in nonhuman primate eyes, providing the possibility of therapeutic application to the clinical practice. CLINICAL RELEVANCE: Occlusion of CNV without direct damage to the sensory retina is useful to preserve visual acuity in patients with exudative age-related macular degeneration. A clinical trial of PDT using ATX S10(Na) is desirable. PMID- 10815158 TI - Retinal toxicity of commercial intravitreal tissue plasminogen activator solution in cat eyes. AB - BACKGROUND: We previously reported retinal toxic reactions in rabbit eyes receiving intravitreal injections of commercial tissue plasminogen activator (tPA) in concentrations greater than or equal to 50 microg/0.1 mL, and recent clinical experience suggests that intravitreal tPA solution may produce toxic effects in human eyes. We therefore investigated the dose-dependent retinal toxicity of intravitreal commercial recombinant tPA solution in cat eyes, which have a vascularized inner retina and vitreous volume similar to that of human eyes. METHODS: Commercial tPA in L-arginine solution was injected into the mid vitreous cavity of normal cat eyes in doses of 25, 50, 75, and 100 microg/0.1 mL and 200 microg/0.2 mL. Control (fellow) eyes received an equal volume of sterile saline solution. After injection, eyes were evaluated by ophthalmoscopy and electroretinography for 14 days and then enucleated for histopathological evaluation. RESULTS: Fundus pigmentary alterations were observed in eyes receiving doses greater than or equal to 50 microg/0.1 mL. Changes were centered in the area around the injection site, and the area's size increased in proportion to the dosage. Mean electroretinography B-wave amplitude measured at 14 days was significantly reduced in eyes receiving greater than or equal to 50 microg of tPA in a dose-dependent fashion. Light microscopy of the involved areas showed loss of photoreceptor elements with necrosis and proliferation of the retinal pigment epithelium. CONCLUSION: Intravitreal injection of commercial tPA solution results in dose-dependent retinal toxicity in cat eyes. CLINICAL RELEVANCE: Because cat eyes are similar to human eyes regarding retinal vascularity and vitreous volume, intravitreal injections of commercial tPA (with L-arginine vehicle) in concentrations greater than 25 microg/0.1 mL are potentially unsafe in human eyes. PMID- 10815159 TI - Matrix metalloproteinases and tumor necrosis factor alpha in glaucomatous optic nerve head. AB - OBJECTIVE: To study expression and location of matrix metalloproteinases (MMPs) and tumor necrosis factor alpha (TNF-alpha) in glaucomatous optic nerve heads, which are known to be secreted in response to a variety of neuronal injury. METHODS: Four postmortem eyes from patients with primary open-angle glaucoma, 7 eyes from patients with normal-pressure glaucoma, and 4 eyes from age-matched normal donors were studied by immunohistochemistry. The sections of the optic nerve heads were examined after immunostaining with antibodies to MMPs (MMP-1, MMP-2, and MMP-3), TNF-alpha, or TNF-alpha receptor 1. RESULTS: The intensity of the immunostaining and the number of stained cells for MMPs, TNF-alpha, or TNF alpha receptor 1 were greater in the glaucomatous optic nerve heads, particularly in eyes with normal-pressure glaucoma compared with age-matched controls. Positive immunostaining was observed in all regions of the glaucomatous optic nerve heads, but most prominently in the postlaminar region. Immunostaining was observed mainly in glial cells and their processes around the axons and blood vessels and in pial septae. CONCLUSION: There is increased immunostaining for MMPs, TNF-alpha and TNF-alpha receptor 1 in the glaucomatous optic nerve head, which suggests increased expression of these proteins in glaucoma and thereby implies a role in the tissue remodeling and degenerative changes seen in glaucomatous optic nerve heads. CLINICAL RELEVANCE: The MMPs and TNF-alpha may be components of astroglial activation that occurs in glaucomatous optic nerve heads. The biological alterations in the expression of these proteins may play a role in the progression of glaucomatous optic neuropathy. PMID- 10815161 TI - Prospective study of alcohol consumption and the risk of age-related macular degeneration. AB - OBJECTIVE: To describe the relationship between alcohol consumption and the incidence of age-related macular degeneration (AMD). METHODS: We conducted a prospective study among female nurses between 1980 and 1994 and among male health professionals between 1986 and 1994. We included 32764 women and 29488 men who were 50 years or older, without a diagnosis of AMD or cancer at baseline, and added additional subjects to the analysis as they reached 50 years of age. Their alcohol intake was assessed at baseline and updated during follow-up evaluations using a validated semiquantitative food-frequency questionnaire. After separate analyses for women and men, pooled estimates of the relationship of alcohol to the risk of AMD were calculated. RESULTS: Age-related macular degeneration associated with a visual acuity loss of 20/30 or worse, including the early and dry and wet types, was diagnosed in 298 women (from 697498 person-years of follow up) and 153 men (229 180 person-years) by 1994, the end of follow-up. After controlling for age, smoking, and other risk factors, the pooled relative risks (RRs) and 95% confidence intervals (CIs) for AMD compared with nondrinkers were 1.0 (0.7-1.2) for drinkers who consumed 0.1 to 4.9 g/d of alcohol; 0.9 (0.6-1.4) for 5 to 14.9 g/d; 1.1 (0.7-1.7) for 15 to 29.9 g/d; and 1.3 (0.9-1.8) for 30 g/d or more. Among women, there was a suggestion of a modest increased risk of the disease in drinkers who consumed 30 g/d or more (RR, 1.5; 95% CI, 1.0-2.4); this was limited to an increased risk of the early and dry form (RR, 2.0; 95% CI, 1.2 3.4). No specific type of alcohol provided protection against AMD. CONCLUSION: This prospective study does not support an inverse relationship between moderate alcohol consumption and risk of AMD. PMID- 10815160 TI - Myocilin Gln368stop mutation and advanced age as risk factors for late-onset primary open-angle glaucoma. AB - BACKGROUND: Juvenile open-angle glaucoma has been found to be associated with molecular defects in the myocilin (MYOC) gene. Most of the defects are missense mutations located in the third exon. The Gln368stop mutation has recently been found in several cases of late-onset primary open-angle glaucoma (POAG). OBJECTIVE: To study the effect of glaucoma risk in a relatively homogeneous genetic population. METHODS: A clinical study was performed in all living members of a 5-generation family. DNA analysis was performed for studying association with genetic markers and identifying the mutation. RESULTS: We identified the Gln368stop molecular defect in 19 patients with POAG, 5 patients with ocular hypertension, and 22 healthy carriers. We compared affected and unaffected carriers based on age at onset and last examination, respectively. Besides the presence of 3 young patients with POAG (<40 years old), the number of glaucomatous patients in the advanced age group increased. CONCLUSIONS: The penetrance of glaucoma increases with age in Gln368stop carriers, but some remain unaffected at advanced age and others are affected at an early age. This suggests that additional risk factors are operating within this family, which may be identified by a genome-wide linkage search in this large pedigree. CLINICAL RELEVANCE: The myocilin Gln368stop mutation shows a good genotype-phenotype correlation and should be investigated in all familiar cases of chronic POAG. This may be important for early diagnosis and periodical checkups of presymptomatic individuals belonging to these families. PMID- 10815162 TI - Molecular genetics of color vision and color vision defects. AB - Color is an extremely important component of the information that we gather with our eyes. Most of us use color so automatically that we fail to appreciate how important it is in our daily activities. It serves as a nonlinguistic code that gives us instant information about the world around us. From observing color, for example, we can find the bee sting on an infant's arm even before it begins to swell by looking for the little spot where the infant's skin is red. We know when fruit is ripe; the ripe banana is yellow not green. We know when meat is cooked because it is no longer red. When watching a football game, we can instantly keep track of the players on opposing teams from the colors of their uniforms. Using color, we know from a distance which car is ours in the parking lot--it is the blue one--and whether we will need to stop at the distant traffic light, even at night, when we cannot see the relative positions of red and green lights. PMID- 10815163 TI - Are the sky and night lights falling? PMID- 10815164 TI - New issues in the management of patients with AIDS-related cytomegalovirus retinitis. PMID- 10815165 TI - Iris varix simulating an iris melanoma. AB - Iris varix is rare and little is known about its clinical characteristics. We treated a thrombosed iris varix that simulated an iris melanoma. A 53-year-old man developed a dark brown iris mass and hyphema in his left eye. Ultrasound biomicroscopy revealed a circumscribed mass of the iris stroma. Because of suspicion for melanoma, it was removed by sector iridectomy. Histopathologic examination disclosed an extensive focus of stromal hemorrhage, partially surrounded by endothelial cells that showed immunoreactivity to vascular markers. The histopathologic diagnosis was thrombosed iris varix. Iris varix is a rare condition that should be included in the differential diagnosis of iris melanoma. PMID- 10815166 TI - Hamartomas of the iris and ciliary epithelium in tuberous sclerosis complex. AB - Astrocytic hamartomas of the retina are the principal ocular manifestation of tuberous sclerosis complex. Iris abnormalities are rare in tuberous sclerosis complex and include focal areas of stromal depigmentation and atypical colobomata. We describe 2 patients who were found on histopathological examination to have lesions consistent with hamartomas of the iris pigment epithelium and ciliary body epithelium. Iris abnormalities, including pupillary irregularities, were noted on clinical examination prior to the development of iris neovascularization in both patients. These observations suggest that iris abnormalities, including atypical colobomas, may be caused by hamartomas of the iris pigment epithelium and ciliary epithelium in some patients with tuberous sclerosis complex. To our knowledge, hamartomas of tissues derived from the anterior part of the neuroectodermal optic cup have not been reported in cases of tuberous sclerosis complex. PMID- 10815167 TI - Iris mammillations as the only sign of ocular melanocytosis in a child with choroidal melanoma. AB - An 8-year-old girl had visual loss in her left eye over 2 months. Ocular examination showed that visual acuity was counting fingers in the left eye. The left iris was moderately pigmented and thickened with numerous confluent, dome shaped elevations on its surface, consistent with iris mammillations arising from ocular melanocytosis. There was total retinal detachment and an inferiorly located large amelanotic choroidal mass compressing the optic nerve. A specimen from a fine-needle aspiration biopsy showed spindle and epithelioid melanoma cells. The eye was enucleated. Pathologic examination showed that the bland melanocytes comprising the anterior border layer of iris formed focal aggregates, corresponding to the iris mammillations observed clinically. The uvea was diffusely thickened. Arising from the posterior choroid and obscuring the optic nerve head was a moderately pigmented spindle and epithelioid cell choroidal melanoma with diffuse lymphocytic infiltration and high mitotic activity. This case demonstrates that iris mammillations can be the initial manifestation of ocular melanocytosis in the absence of scleral pigmentation. PMID- 10815168 TI - Eyelash formation secondary to latanoprost treatment in a patient with alopecia. PMID- 10815169 TI - Sarcoid inflammation and obstruction of the nasolacrimal system. PMID- 10815170 TI - Pneumatic retinopexy in a progressive rhegmatogenous retinoschisis retinal detachment. PMID- 10815171 TI - Diffuse unilateral subacute neuroretinitis in China. PMID- 10815172 TI - Ocular manifestations of alkaptonuric ochronosis. PMID- 10815173 TI - Purtscher retinopathy and necrotizing vasculitis with gemcitabine therapy. PMID- 10815175 TI - Accurately Analyzing Aqueous Flare Data. PMID- 10815174 TI - Optic disc sarcoid granuloma. PMID- 10815176 TI - Accurately Analyzing Aqueous Flare Data. PMID- 10815177 TI - Primary Intraocular Lymphoma With a Low Interleukin 10 to Interleukin 6 Ratio and Heterogeneous IgH Gene Arrangement. PMID- 10815178 TI - Primary intraocular lymphoma with a low interleukin 10 to interleukin 6 ratio and heterogeneous IgH gene arrangement. PMID- 10815179 TI - The BB gun is equivalent to the airsoft gun in the Japanese literature. PMID- 10815180 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815181 TI - Health outcomes among patients treated by nurse practitioners or physicians. PMID- 10815183 TI - Health outcomes among patients treated by nurse practitioners or physicians PMID- 10815182 TI - Health outcomes among patients treated by nurse practitioners or physicians PMID- 10815184 TI - Mercury(II) tungstate from neutron powder data PMID- 10815185 TI - Fe2P2O7(H2O)2 PMID- 10815186 TI - Two forms of aluminium phosphate tridymite from X-ray powder data PMID- 10815187 TI - Strontium chromium(II) diphosphate, SrCrP2O7 PMID- 10815188 TI - The sodium salt of a tris(tridentate anion)gadolinium(III) complex: pentasodium bis PMID- 10815189 TI - Reinvestigation of bis(trimethylsilyl)mercury PMID- 10815190 TI - A novel trinuclear palladium cluster compound: di-mu 3-chloro-tris PMID- 10815191 TI - Carbonyl PMID- 10815192 TI - Poly PMID- 10815193 TI - Aqua(n-hexyl)[3,3'-(propane-1,3-diyldinitrilo)bis(butan+ ++-2-one) dioximato kappa 4N]cobalt(III) perchlorate. PMID- 10815194 TI - [N,N'-bis] PMID- 10815195 TI - One-dimensional polymeric chain structure of bis(aniline)dithiocyanatocadmium(II) PMID- 10815196 TI - Salts of two pentahalo(N-donor)bismuthate(III) anions: PMID- 10815197 TI - Acetatodiaqua PMID- 10815198 TI - Sodium mycophenolate. PMID- 10815199 TI - [Sm] PMID- 10815201 TI - Mixed-chalcogenide double-butterfly complex PMID- 10815200 TI - A host-guest complex of diaquabis[1-hydroxy-2(1H)-pyridinethionato O,S]magnesium(II ) and 2,2'-dithiobis(pyridine N-oxide). PMID- 10815202 TI - Three sulfur-containing diborane(4) compounds PMID- 10815204 TI - Chelidamic acid monohydrate: the proton complex of a multidentate ligand PMID- 10815203 TI - [New derivatives of pantolactonyl N-phthalimido-2-arylpropanoate]. AB - We present the crystal and molecular structures of two new N-phthalyl-3-amino-2 arylpropionic acid pantolactonyl ester derivatives with 4-fluorophenyl and 3,4 dimethoxyphenyl as the aryl group, 2,3,4,5-tetrahydro-4,4-dimethyl-2-oxofuran-3 yl 3-phthalimido-2-(4-fluorophenyl)propanoate, C23H20FNO6, and 2,3,4,5-tetrahydro 4,4-dimethyl-2-oxofuran-3-yl 3-phthalimido-2-(3,4-dimethoxyphenyl)propanoate ethyl acetate hemisolvate, C25H25NO8.0.5C4H8O2. This structural study confirms the S configuration of the C2 and validates the stereospecificity of our synthesis strategy. PMID- 10815205 TI - cis-Verbenol. PMID- 10815206 TI - Neutral bis(ethylenedithio)tetrathiafulvalene at 100 K PMID- 10815207 TI - The antiestrogen [2-(4-benzylphenoxy)ethyl]diethylammonium chloride. PMID- 10815208 TI - Flavone-3'-sulfonamide. PMID- 10815209 TI - Methyl 1-(4-chlorobenzyl)-2-(4-methylpiperazin-1-yl)-1H-benzimidazole-5- carboxylate hemihydrate. PMID- 10815210 TI - The fused heterocycle 2-benzyl-3-methyl-1-phenyl-4-phenylsulfonyl-2H,4H-pyrrolo PMID- 10815211 TI - 4-Fluoro-2-(phosphonomethyl)-benzenesulfonic acid monohydrate PMID- 10815212 TI - 2,4,6-Tris(methylthio)-1,3,5-triazine,3-methyl-4,6-bis(methylthio)-1,3,5- triazine-2(3H)-thione,1,3-dimethyl-4-methylthio-1,3,5-triazine-2,6(1H,3H)- dithione and 1,3,5-trimethyl-1,3,5-triazine-2,4,6(1H,3H,5H)-trithione PMID- 10815213 TI - N-carboxy-L-phenylalanine anhydride. PMID- 10815214 TI - Beta-pyridinium dichloroiodide PMID- 10815215 TI - Two 5-oxa-2,6-diazaspiro PMID- 10815216 TI - 4-[1-(phenylsulfonyl)indol-3-yl]-3a,4,5,9b-tetrahydro-3H-cyclopenta[c] quinoline. PMID- 10815217 TI - 1:1 hetero-assembly of 2-aminopyrimidine and (+)-camphoric acid PMID- 10815218 TI - (8-Dimethylamino-1-naphthyl)dimethylammonium 3-carboxy-1,4,5,6,7,7 hexachlorobicyclo PMID- 10815219 TI - (3R,4S)-3-phenyl-1-[(R)-1-phenylethyl]-4-[(R)-1-phenylethyliminomethyl] azetidin 2-one. PMID- 10815221 TI - 1,2-Ph2-9-I-1,2-closo-C2B10H9 PMID- 10815220 TI - (E)-6-methoxy-3-(alpha-methoxybenzylidene)benzo PMID- 10815223 TI - N,N'-bis(2-tosylaminobenzylidene)benzene-1,2-diamine PMID- 10815222 TI - 7-Iodo-8-aza-7-deaza-2'-deoxyadenosine and 7-bromo-8-aza-7-deaza-2' deoxyadenosine. PMID- 10815224 TI - Ethyl 3-(2'-deoxyuridin-5-yl)-3-hydroxy-2-iodopropanoate, a nucleoside analogue. PMID- 10815225 TI - Ammonium N-acetyl-L-threoninate and methylammonium N-acetyl-L-threoninate. PMID- 10815226 TI - A new 'hydrogen-bond rule' applied to the structure of L-seryl-L-alanine and pairs of dipeptide retroanalogues. PMID- 10815227 TI - 1-(3-Methoxybenzoyl)-3,3-diethylthiourea PMID- 10815228 TI - Triterpenoide. XIX. 3 beta-Hydroxy-11-oxo-18 alpha-olean-12-en-28-saure methylester und 3,11-dioxo-18 alpha-olean-12-en-28-saure-methylester PMID- 10815229 TI - 1-[5-[2-(trifluoromethyl)phenyl]-1,3,4-thiadiazol-2-yl]guanidinium chloride. PMID- 10815230 TI - Epidemiology of cypress allergy. PMID- 10815231 TI - The natural history of mountain cedar pollinosis. AB - During the winter months, pollen from the mountain cedar (MC) (Juniperus sabinoides) causes severe respiratory tract allergy in central Texas. We have been impressed with the fact that many of our MC-allergic patients had only allergic rhinitis and were only sensitive to MC pollen. We therefore studied 234 unselected MC-allergic patients at the end of the MC season. The main criteria for inclusion into the study was a bona fide history of MC pollinosis confirmed by a positive skin test. All patients completed a detailed history questionnaire, were prick skin tested by using 1:20 w/v, extracts, and had a total IgE determined with commercially available reagents (PRIST). Thirty-four percent of patients were found to be allergic only to MC and 66% were allergic to MC and other aeroallergens. As a group, patients allergic only to MC had significantly lower total IgE levels (84 IU/ml vs. 360, p < 0.001) (normal < 180), required much longer exposure intervals to cedar pollen before developing MC pollinosis (14.4 yr vs, 5.69, p < 0.001), had less of a family history of allergic disease (43% vs. 88%, p < 0.001), had less of a history of asthma or eczema (11% vs. 39%, p < 0.001), and developed allergic disease at a later age (39 yr vs. 13, p < 0.001) when compared to patients with multiple allergies. These results suggest that the MC pollen may be unique in causing allergic rhinitis in patients who have no other sensitivities. A possible explanation may lie in the carbohydrate nature of the main allergen of the MC pollen, which may facilitate allergen transport through the respiratory tract mucosa and subsequent sensitization. PMID- 10815232 TI - Environment and sensitization to cypress in Israel. AB - Cypresses are part of the Israeli landscape, as well as they are over most of the Mediterranian area. Pollen dispersal starts in January and peaks in March-April. Assessment of hay-fever patients showed a prevalence of 24%-32% (depending on the region) sensitization to Cypress (Cupressus sempervirens) pollen extracts. 13% of all those who are sensitive to Cypress were monosensitized. They are symptomatic from February till April, when 70% of them have rhinitis, 30% also asthma and 18% conjunctivitis. In the monosensitized patients, anti-cypress immunotherapy was successful. PMID- 10815233 TI - Pathogenesis of allergic diseases: interactions between pollutants and pollens really important? AB - Nowadays, the prevalence of atopic diseases in so-called developed countries is estimated to exceed 30%. Furthermore, it may reach over 50% in two generations. Based on such facts, the so-called "atopic predisposition" can not be defined as an abnormal genotype consisting of certain "atopic gene(s)" possessed by a minority of unfortunate people. Rather, the gene(s) that cause atopic diseases reside in the common human gene repertoire, and several environmental factors would cause the overexpression of some constitutive gene(s), leading to the development of atopic diseases. The author considers that overexpression and production of Th2 cytokines, especially IL-5, may be a key event. The recent prevalence of atopic diseases in developed countries may be mediated by a change of Th2 polarization due to modern retrogression of environmental factors such as bacterial and viral infections that favor Th1 polarization. The "atopic trait" might be actually an "atopic phenotype" rather than an "atopic genotype". In other words, the atopic trait seems not to be a genotype that decides the development of an atopic disease on an all or nothing basis, but a phenotype that determines the susceptibility to the disease. Of course, the familial nature of atopic diseases is undeniable, but this does not necessarily indicate the genetic nature of atopic diseases. For example, if a parent suffers from tuberculosis, the possibility that the children will develop tuberculosis markedly increased. However, tuberculosis is a truly infectious disease. In this article, several environmental factors those may affect the recent sharp increase of atopic diseases are discussed. PMID- 10815234 TI - Analysis of Japanese monkeys (Macaca fuscata) and dogs with naturally occurring Japanese cedar (Cryptomeria japonica) pollinosis. AB - The natural occurrence of CJ pollinosis has been found in Japanese monkeys and dogs. These animals have reactivity to CJ major allergens. Monkeys and dogs can serve as a suitable animal model in the study of CJ pollinosis. PMID- 10815235 TI - Recombinant cypress allergens. AB - Cedar/cypress/juniper pollen allergy is probably one of the most common worldwide pollen allergy. Purification, identification and characterization of allergens increased the knowledge about the IgE reactivity of allergic subjects toward defined allergenic components. Molecular biology give the opportunity to produce the allergens in virtually unlimited reproducible amount, to further characterize the IgE reactivity of the allergenic molecule, and finally how this reactivity can be modified for therapeutic purposes. The DNA recombinant technique has been applied to the study of cedar/cypress/juniper pollen allergens and several molecules have been cloned. These molecules can be divided in four groups depending on their biologic activity: he Pectate lyases (rCry j 1, rCha o 1, rJun a 1); the Polygalacturonase (rCry j 2, rCha o 2); the Thaumatin-like pathogenesis related protein (rJun a 3); the Calcium-binding protein (rJun o 2). Main immunobiologic features of these cedar/cypress/juniper recombinant allergens will be summarized. PMID- 10815236 TI - [Allergy to cypress pollen: preparation of a reference and standardization extract in vivo]. AB - Development of Cypress allergy frequency led to the standardization of commercial cypress extract used for diagnosis and immunotherapy. Previous in vitro studies on two cypress pollen species (Cupressus sempervirens and Cupressus arizonica) allowed us to produce an allergenic solution composed by a mixture of both extracts for in vivo standardization. Dilutions of this allergenic solution were tested by prick-test on 44 patients with clinical allergy to cypress pollen to define the dilution that corresponds to a 6 mm wheal conformed to the definition of 100 IR. The mixture of the two major species found in France is justified by the in vitro study results. Extracts revealed complementary allergenic composition: Cup sempervirens showed a wider diversity of allergens whereas Cup arizonica showed a higher content of the major 43 kDa allergen. Thus, according to in vivo analysis, we are able to produce a standardized extract of Cypress pollen expressed in IR. PMID- 10815237 TI - [Juniperus ashei: the gold standard of the Cuppressaceae]. AB - The non-standardized Cupressus sempervirens allergen extract currently available for the diagnosis of cypress allergy has a low level of activity. The search for an active material consisted of in vitro and in vivo comparison of three Cupressaceae pollen extracts: Cupressus sempervirens (Cs), Cupressus arizonica (Ca) and Juniperus ashei (Ja) (synonyms: Juniperus sabinoides and Mountain Cedar). These 3 trees belong to the same botanical family of Cupressaceae. While Cs and Ca are commonly encountered in Mediterranean regions, Ja is only present in Europe in the Balkans, but is a major cause of allergy in the USA. In vitro, with a similar protein content, the allergenic properties of Ja extract are 20 Fold higher than those of Cs and 11-fold higher than those of Ca. IgE immunoblotting revealed 14, 42 and 70 kDa allergens common to all 3 extracts. The inhibition curves of the 3 extracts were more than 88% parallel. A significant correlation was observed between serum specific IgE titres for Ja and Cs in 23 patients (r = 0.916; p < 0.001). In vivo, in 23 patients with cypress allergy, the mean diameter of the prick test papule at 1/20 W/V of Ja (8.3 mm) was greater than that of the Cs papule (6.3 mm) (p = 0.001) and the Ca papule (6.7 mm) (p < 0.001). Correlations between cutaneous responses to Cs and Ja (r = 0.629; p = 0.002), and to Cs and Ca (r = 0.75; p = 0.001) were significant. These results demonstrate the intense cross-reactivity between Cs, Ca and Ja. The allergenic potency of the Ja extract is superior to that of Cs and Ca extracts, both in vitro and in vivo. This superiority is correlated with a high concentration of the major allergen, Jun a 1. The non-standardized The now standardized extract of in vitro ashei pollen therefore represents an effective and documented solution for identification, and probably for treatment, of Cupressaceae pollen allergy. PMID- 10815238 TI - [Cypress arizona: allergic extracts with a diagnostic purpose]. PMID- 10815239 TI - Concentration of airborne pollen from Cupressaceae in Lisbon. AB - The concentration of airborne pollen from Cupressaceae was regularly monitored++ in Lisbon during 1997 and 1999, and the phenology of flowering cypress was studied in several species of the genus from 1992 to February 2000. Both methods showed a peak of pollen abundance during the month of February, with the airborne pollen concentration decreasing strongly to March and April. The results obtained are in accordance with the literature for the Mediterranean area, but in Lisbon the peak started and finished earlier than in other Mediterranean towns already studied. PMID- 10815240 TI - Cupressacae: comparative pollen counts from the Athens metropolitan area 10 years apart (1987-89 and 1997-99). PMID- 10815241 TI - [Skin sensitivity to cypress pollen in Morocco: multicenter study]. PMID- 10815242 TI - Cypress in Italy: landscape and pollen monitoring. AB - In Italy the Cupressaceae pollen has been monitored by the Italian Aeroallergen Network (IAN) in collaboration with the Italian Research Council (CNR) since 1988. The measurements carried out over the last decade show a positive trend of the seasonal emission of the cypress pollen, whose flowering peak occurs in Italy from mid-February to mid-March. During the pollination period the cypress pollen represents 10% to 40% of the total pollen, showing a high variability of its annual production. PMID- 10815243 TI - Cypress pollen counts in Israel. AB - In accordance with the terms of the modern world, this presentation is based on the WWW, with a small change in interpretation: WWW stands for Who? When? and Where? PMID- 10815244 TI - Cupressaceae in Tirana (Albania) 1996-1998 aerobiological data and prevalence of Cupresaceae sensitization in allergic patients. AB - Plants of Cupressaceae family are quite widespread in Albania. Aerobiological data show that 43% of the total airborne pollens per year come from this family. The highest concentrations of Cupressaceae pollen were found from February to May, while low quantities (till 32 p/m3) were present form November to January. From 110 patients with respiratory allergies and positive-SPT, only three subjects (2.7%) were SPT-positive to Cupressaceae. All the three were polysensitized. PMID- 10815245 TI - Cypress pollen: botanic aspects in fourteen cypress species and prospects for research. AB - The two Research Institutes, the CNR (National Research Institute, Italy) and the INRA (Institut National de Recherche Agronomique, France) which have been collaborating on the problem of cypress canker from Seiridium cardinale since 1975 in the context of EU projects, have measured in the SEM the pollen size of fourteen species of cypress, determining the differences between the maximum diameter of 100 granules per species and noting differences in pollen colour. A research strategy based on the use of material previously selectively bred for canker resistance is proposed to maintain the cypress in its Mediterranean habitat with a reduced risk of pollinosis. For the success of the project it is stressed that close collaboration between Forest research institutions and the medical profession will be necessary. PMID- 10815246 TI - [Specific desensitization to cypress]. PMID- 10815247 TI - [Cupressaceae: genera and species]. PMID- 10815248 TI - [Natural environment, geographic distribution and ecology of cypress]. PMID- 10815249 TI - [Allergy to cypress: clinical aspects]. PMID- 10815250 TI - New clinical data and therapeutic prospects in Cupressaceae pollen allergy. AB - The authors emphasize, thanks to their clinical data over the last 20 years: The steady increase of allergy to C. sempervirens pollen compared to other pollinosis and to atopy. The frequency of cross-reactivity between C. sempervirens and other pollens belonging to Cupressaceae and Taxodiaceae families, in order, in the absence of a very good standardization and of a purification of this pollen, to try to undertake a sound specific immunotherapy using the most cross-reactive pollen mixed with C. sempervirens extracts. As a matter of fact the partially purified extracts of C. japonica gave the higher incidence (91%) of cross reactivity. Consequently, a mixture of C. sempervirens and C. japonica extracts was used since 1993 with good results in a double blind study. PMID- 10815251 TI - [AFEDA (Association Francaise d'Etudes des Ambroisies), its history, actions, and politics]. PMID- 10815252 TI - How to reduce air pollution by Cupressus pollen? AB - Cypress trees are among the most famous trees of the Mediterranean region. The origin of Cupressus sempervirens is apparently in the East Mediterranean. A nice description of the site of origin of Cupressus was provided by the Roman philosopher Plinius: "cypress trees sprout spontaneously on Mt. Ida on the island of Crete, whenever the soil there is disturbed". The aesthetic shape of the trees, the quality of their timber and their adaptation to harsh environmental conditions has attracted people. Certain varieties of the tree (e.g., var. Pyramidalis) were spread rapidly all over the Mediterranean and since the 16th century were introduced into the Western Hemisphere and other parts of the world. During historical times, Cupressus sempervirens was so widely used in the East Mediterranean countries that it reached the brink of extinction. Nowadays it is planted on a large scale as an ornamental plant as a windbreak and as a forest tree. Such plantations have various impacts on their environment, many positive ones, some negative ones, and some disastrous ones. This last aspect is where cypresses come in. PMID- 10815253 TI - [History of allergy to cypress pollen]. PMID- 10815254 TI - [Cypress situation in the Mediterranean region]. PMID- 10815255 TI - [Alternative trials to the urban use of cypress]. PMID- 10815256 TI - [Methodology of exercise test in patients with heart failure. Maximal test and submaximal test]. PMID- 10815257 TI - [Methodology of exercise test in patients with heart failure. The cardiopulmonary test: mode of execution, instrumentation, protocols, reproducibility]. PMID- 10815258 TI - [Methodology of exercise test in patients with heart failure. Serial tests: rationale and interpretation of results]. PMID- 10815259 TI - [Physiopathology of exercise. Cardiac adaptation]. PMID- 10815260 TI - [Physiopathology of exercise. Hormonal and metabolic adaptations]. PMID- 10815261 TI - [Physiopathology of exercise. Vascular adaptations]. PMID- 10815262 TI - [Physiopathology of exercise. Respiratory adaptations]. PMID- 10815263 TI - [Physiopathology of exercise. Muscular adaptations]. PMID- 10815264 TI - [Objective evidence of exercise intolerance. What parameters?]. PMID- 10815265 TI - [Objective evidence of exercise intolerance. Relationship with clinical variables]. PMID- 10815266 TI - [Objective evidence of exercise intolerance. Hypothesis on mechanisms responsible for symptoms]. PMID- 10815267 TI - [Objective evidence of exercise intolerance. Differential diagnosis of cardiac and pulmonary dyspnea]. PMID- 10815268 TI - [Ergometric test and level of daily physical activity in chronic heart failure]. PMID- 10815269 TI - [Exercise test in the evaluation of the efficacy of drug therapy]. PMID- 10815270 TI - [Exercise test in the evaluation of the efficacy of non-drug therapy]. PMID- 10815271 TI - [Exercise test in prognostic stratification]. PMID- 10815272 TI - [Evidence-based medicine: reality and illusions. Extension of epistemological reflexions]. AB - Evidence-based medicine (EBM) is a cultural and methodological approach to clinical practice helping to make decisions based on clinical expertise and an intimate knowledge of the individual patient's situations, beliefs, and priorities useful for the analysis of clinical research. As such, it can be considered the scientifically grounded art of medicine, as it appears to be an emerging paradigm of scientifically based clinical care. It de-emphasizes intuition and unsystematic clinical experience as grounds for medical decision making and stresses the rigorous and formal analysis of evidence from clinical research. EBM converts the abstract exercise of reading and appraising the literature into the pragmatic process of using the literature to benefit individual patients, while simultaneously expanding the clinician's knowledge base. On EBM grounds, clinical, practice guidelines, pathways and algorithms or instructions can be developed with the aim of solving a problem or accomplishing a task. Nonetheless in these processes the theory of EBM shows internal and external bias. Among internal bias, economic-based interest may influence the development and diffusion of research and its results. In addition "systemic review" may be incorrectly guided, the quality filters of the literature can be inappropriately applied, the choice criteria can be only based on the positive results of evidence, but according to modern epistemology, it will be helpful for clinicians to know when their uncertainty stems from gaps between positive and negative evidence. Another bias is the difficulty to convert EBM into clinical practice recommendations. EBM set movement has shown that it is nearly impossible to make recommendations that are appropriate in every situation. Epistemological approach identifies external "bias" of EBM. It is consistent with the theory of "fact" as human construction. Every human fact can historically fade and then be restored according to new paradigms. EBM is a "fact" and its theory can be changed or removed every time by relevant new or emerging evidence approaching the development of up-to-date decision-making aids and so on, irrespective of how much previous processing of the evidence has taken place. Then EBM cannot be evaluated as the scientific "totem" of the third millennium, neither as the clinical digest of medical literature. Searching for clinical evidence, in fact, requires a great awareness of both the advantages and limitations of increasing bias. Clinicians are looking for new strategies to apply to diagnostic and therapeutic pathways and for the steps where EBM could be addressed when showing the full validity. PMID- 10815273 TI - [Right ventricular thrombosis in arrhythmogenic cardiomyopathy. A case report]. AB - Echocardiographic demonstration of right ventricular thrombosis is relatively common in pulmonary embolism. There are also reports of right ventricular thrombi in patients affected by right myocardial infarction or dilated cardiomyopathy. In arrhythmogenic right ventricular cardiomyopathy single or multiple aneurysms are often present in the right ventricular free wall. These hypoakinetic areas represent a site for potential development of thrombi especially in advanced disease states. In the literature a single case of a patient affected by arrhythmogenic right ventricular cardiomyopathy with right heart failure and atrial and ventricular thrombi is reported. We report a case of arrhythmogenic right ventricular cardiomyopathy with a right ventricular thrombus located inside a single apical aneurysm in the presence of normal right ventricular systolic function. PMID- 10815274 TI - [Heart arrest caused by ventricular fibrillation in acute biliary pancreatitis: a case report and etiopathogenetic hypothesis]. AB - Acute pancreatitis and/or gallbladder disease precipitating angina pectoris, or producing arrhythmias or ST-T wave changes on electrocardiography in the absence of coronary artery disease, have frequently been mentioned. Many attempts have been made to explain the etiology of these findings. To the authors' knowledge, this report is the most dramatic expression of pathogenetic mechanisms which are still considered hypothetical. PMID- 10815275 TI - [Images in cardiovascular medicine. Appearance is deceptive]. PMID- 10815276 TI - [Blocking of aldosterone receptors reduces the risk of events in patients with severe cardiac failure]. PMID- 10815277 TI - [Dofetilide in patients with chronic heart failure and left ventricular dysfunction]. PMID- 10815278 TI - [Comparison of candesartan, enalapril and their combination in congestive heart failure]. PMID- 10815279 TI - [Primary angioplasty is superior to thrombolytic therapy in also reducing long term clinical events after acute myocardial infarct]. PMID- 10815280 TI - [Physical exercise and survival after myocardial infarction]. PMID- 10815281 TI - [Use of estrogen-progestins in secondary prevention]. PMID- 10815282 TI - [More on the protective role of wine]. PMID- 10815283 TI - [More on the protective role of wine]. PMID- 10815284 TI - [Multiple sclerosis: a stake for understanding by researchers and a therapeutic challenge for doctors]. PMID- 10815285 TI - [Research on multiple sclerosis and tissue banks]. AB - Tissue banks are of major importance in research on human tissues, in particular as regards the furthering of our knowledge on multiple sclerosis (MS). Individuals who wish to make a 'donation of their brain' for autopsy, or pathologists in possession of biopsy specimens that have not been utilized for diagnosis provide the necessary material for investigation by research teams. In addition to their technical aspects, brain tissue banks provide information and aid in promoting research. Their functioning, usually supported by patient associations, has encountered certain difficulties. At present, it is challenged by a decrease in the number of autopsies. PMID- 10815286 TI - Methodological problems in evaluating efficacy of a treatment in multiple sclerosis. AB - Multiple sclerosis is currently an incurable disease and there is much interest in developing new disease-modifying treatments and testing them in clinical trials. Because therapies are essentially partially effective, neurologists have to develop ways to assess whether the medication they test slows down the course of the disease. Promising treatments selected for Phase III clinical trials are first tested in Phase I/II studies, where preliminary evidence of efficacy is demonstrated in the absence of severe adverse events. Phase III clinical trials are designed to detect significant differences between responses to active therapies and a placebo or a marketed drug. The selection and usage of instruments to detect changes in patients is probably the most important issue in their design. The purpose of this paper is to discuss methodological problems of evaluating drug efficacy in phase III MS trials, including the characteristics of the ideal instrument to measure progression of the disease, the strengths and limitations of existing clinical, radiologic and laboratory outcome measures. PMID- 10815288 TI - [Cognitive and emotional disorders in multiple sclerosis. Can a management strategy be envisioned?]. AB - There is a lack of precise data concerning the natural history of cognitive disorders in multiple sclerosis (MS), but recent neuropsychological studies have demonstrated that the incidence of such disorders in MS appears to be frequent (40-65% of cases), and have shown in particular that recent memory, conceptual reasoning, attention, executive functions, visuospatial perception and information processing speed are negatively affected. In contrast, language functions, general intelligence and implicit memory appear to be relatively well preserved. Although the presence and the degree of cognitive disorders does not seem to be directly linked to disease duration or to the extent of physical disability, the relationship between cognitive decline and brain lesions detected by magnetic resonance imaging (MRI) is still a subject of discussion. The prevalence of emotional and affective disorders is difficult to estimate. Their frequency has only rarely been investigated, and the lack of data on the natural history of these disorders and those factors which they have in common (the psychosocial consequences of this chronic and disabling disease, cognitive impairment, and brain lesions) further complicate the determination of treatment strategy. The adoption of appropriate strategies could limit the negative impact of this disease on the social functioning of MS patients. PMID- 10815287 TI - [Is immunosuppression a future therapeutic strategy for multiple sclerosis?]. AB - Although in previous studies no clear demonstration was found of the efficacy of azathioprine, cyclophosphamide and methotrexate as immunoprophylactic agents in cases of multiple sclerosis (MS), over the past five years a number of well designed clinical trials utilizing immunosuppressive and immunomodulatory agents have shown partial efficacy regarding the drugs involved, but they have not been able to determine in what way these drugs can modify the natural course of this disease. Among the immunosuppressors, mitoxantrone is of particular interest as during the past two years three controlled trials have taken place in Europe and have demonstrated its efficacy both as regards clinical (frequency of symptoms, progression of the disease) and magnetic resonance imaging (MRI) criteria. Due to its potentially severe cardiotoxicity related to total cumulative dose, mitoxantrone is only prescribed for a limited period, and its use is limited to selected patients with a high relapse rate and incomplete remission, or to those who do not respond to beta-interferon treatment. The immunomodulatory agents have less immediate efficacy, but because they are well tolerated they can be used early in the course of the disease and over a prolonged period of time. The beta interferons (1a or 1b) have been given market approval for use in the treatment of MS: three large, randomized, double-blind studies have demonstrated their capacity to reduce by 30% the frequency of symptoms and the appearance of disabilities associated with relapse and with the progression of the disease. Glatirameracetate or copolymer 1, which is available in France (ATU), has been found to reduce the frequency of relapse by 30%. It constitutes an alternative immunomodulatory treatment for relapsing-remitting patients without major functional disabilities and who suffer from severe side effects with beta interferon treatment. In the future, the early use of immunomodulatory agents and therapeutic drug combinations may be introduced. Therapeutic trials are currently in progress to determine the viability of this approach. PMID- 10815289 TI - [Can the prognosis of multiple sclerosis be predicted?]. AB - Though the global course of the disease is well known when considering large cohorts of patients, it is rather difficult to make a prognosis for a given patient. It is essential however to predict the course of the disease as early as possible in order to be able to give the patient the right treatment at the right time. Age at disease onset is one of the most important clinical predictive factors, as this has been proved by various analyses. The earlier the disease onset, the better the prognosis. The inaugural symptom of optic neuritis, the long interval between the first two relapses, a remitting form at onset are predictive factors of a good evolution. A normal brain magnetic resonance imaging performed on the occasion of a first isolated sign of the central nervous system is the marker of a good prognosis. Conversely, the presence of oligoclonal bands in the cerebrospinal fluid and the increase of IgG index increase the risk of occurrence of a second relapse. The new techniques of magnetic resonance imaging and the exhaustive initial clinical examination using several classifications of disability will allow a better prognosis for a given patient. PMID- 10815290 TI - [Economic repercussions of multiple sclerosis]. AB - Following a definition and critical examination of the various concepts utilized in health economy, the authors have made an analysis, based on a review of the literature available on this subject, of the economic repercussions of multiple sclerosis (MS) on the different individuals and organizations involved (social services, the patient, the patient's family). They have also reported the preliminary results of a French study carried out on a sample population of 157 MS patients undergoing treatment at four different neurological departments in the Nord--Pas de Calais region. PMID- 10815292 TI - The pharmacy world of science and practice PMID- 10815291 TI - [Magnetic resonance imaging in multiple sclerosis]. AB - The contribution of magnetic resonance imaging techniques to the clinical prognosis of multiple sclerosis. Magnetic resonance imaging (MRI) is a diagnostic technique with a high sensitivity for the detection of lesions, but with a poor pathological specificity. In the case of multiple sclerosis (MS), the improvement of diagnostic efficacy depends on a careful analysis of the clinical presentation and the use of increasingly stringent MRI criteria aimed at improving the specificity of the conventional MRI T2 sequences. New sequences such as fast spin echo (also called turbo spin-echo) and FLAIR (fluid attenuated inversion recovery, a method derived from inversion recovery) have improved the visualization of lesions. MRI can under certain conditions be used to monitor the evolution of MS. Acute-phase monitoring is focused on observed changes in disease activity such as the appearance, recurrence or extension of lesions after i.v. injection of contrast medium, i.e., gadolinium (Gd)-enhanced MRI. In the chronic phase, the lesions is the aspect used as the monitoring criterion. However, MRI is still only a secondary criterion in phase III therapeutic trials due to its insufficient correlation with the disability. In neurological daily practice, conventional MRI is only of limited interest at the individual level in patient follow-up, as its prognostic value is poor. Moreover, the difficulty in determining the lesion load can only be excluded in the context of clinical trials, in which certain methodological precautions are taken. This is why techniques other than MRI are being investigated to obtain a better correlation with the clinical course of the disease, for instance the quantification of 'black holes' on T1 weighted images, and the measurement of cerebral and spinal atrophy. Adapted MRI techniques allow a weighted signal to be obtained via the movement (diffusion imaging), by the complexity of the molecular structure (magnetization transfer imaging), by chemical shift (spectroscopic imaging), or by local oxygenation (functional MRI). These new MRI techniques allow a more precise assessment of the pathological mechanisms involved in MS, such as edema, blood brain barrier break-down, demyelinisation, gliosis, cellular infiltration and axonal loss; they provide a better means of establishing the correlation between clinical impact and the destructive nature of the MS lesion. The importance of axonal loss has recently been confirmed in MS by analyzing MRI spectroscopic and neuropathological findings. In addition to magnetization transfer imaging, MR diffusion imaging and functional MRI are being intensively studied in order to assess their contribution to the study of reversibility of the degenerative process. PMID- 10815293 TI - A nation-wide study on the practice of euthanasia and physician-assisted suicide in community and hospital pharmacies in The Netherlands. AB - PURPOSE: In recent years the attitudes and practices of euthanasia (E) and physician-assisted suicide (PAS) among health care workers have been evaluated in different countries. There is, however, still an information gap on the role of pharmacists in these matters. The aim of our study was to study the attitudes and practices of E and PAS among pharmacists. METHODOLOGY: We conducted a nation-wide survey into the practice, including adherence to guidelines, of E and PAS in community and hospital pharmacies (CP and HP) in the Netherlands. Furthermore, the attitude of community pharmacists concerning this subject was evaluated. Anonymous questionnaires were sent to a random sample of 50% (n = 755) of all CP and to all (n = 101) HP. RESULTS: The response rates were 52% and 51% for CP and HP, respectively. Most of the CP-respondents (95%) agreed with the concept of E and PAS and would dispense drugs for these purposes. When the data were extrapolated to all pharmacies, there were 1351 and 340 dispensing of drugs for E and PAS per year in CP and HP (94% of all requests), respectively. In most cases the pharmacist had been notified of the condition of the patient (CP: 93%, HP 87%) and a written request was obtained (CP: 74%, HP: 79%). The drugs were often handed personally to the physician (CP: 98%, HP: 86%). Involvement of pharmacy technicians was more common in HP than in CP (31% versus 6%). The most frequently dispensed drugs were muscle relaxants with barbiturates (CP: 47%, HP 71%), barbiturates only (CP: 19%, HP: 6.1%), and muscle relaxants with benzodiazepines (CP: 14%, HP: 7.6%). CONCLUSION: Most pharmacists are supportive of E and PAS and are prepared to fill prescriptions written for these purposes. PMID- 10815294 TI - Ethics applied to pharmacy practice. AB - This article tries to develop an ethical reasoning that can be applied to (the practice of) pharmacy. Only general principles, based on accepted values in western society, lead to guidelines for ethical behaviour. Such essential values are personal autonomy, democracy and solidarity. The principle of nonmaleficience can be derived from these. Results of this analysis can be applied to health care and pharmacy practice. Subchapters deal with questions such as budget limitations and the autonomy of the patient versus that of the care provider. It concludes that protocols are important tools for ethical behaviour in every day practice. The ethical problem appears to be the unequal access to the health care system. An analysis of pharmaceutical care in the light of ethics can help to formulate the pharmacist's responsibilities. The principle of nonmaleficence is strongly connected to the pharmacy profession. Pharmacists should focus more on possible negative outcomes of pharmacotherapy. Monitoring the patient's medication, identification and prevention of possible adverse effects, medication surveillance, proper communication and information about the use of medicines are therefore priority items within our profession. A definition of target groups for pharmaceutical care will facilitate this task. A suggestion for a general code of ethics for pharmacists is proposed and compared with the code of ethics as currently accepted by the International Pharmaceutical Federation (FIP)-council. PMID- 10815295 TI - Diary cards and the assessment of cough symptoms in community pharmacies. AB - The aim of this study is to compare cough symptom assessment and medication efficacy as recorded in a diary card with that obtained through direct questioning by a pharmacist. Clients visiting six pharmacies, purchasing a cough medication and meeting the inclusion criteria were recruited. They were asked to fill out diary cards to obtain a daily retrospective assessment of symptoms and medication efficacy. After three days the subjects returned to the pharmacy and were asked to recall their symptoms through a structured questionnaire administered by the pharmacist. 48 clients were recruited to the study and of these 44 (92%) completed the study. A good correlation was obtained between ratings of medication efficacy as recorded by diary cards and recall. Symptom assessment also showed a statistically significant correlation for days two and three only. For studies assessing symptoms over a 48 hour period, detailed post study assessments may not be required. The study provides further evidence for the viability of clinical trials conducted through community pharmacies. PMID- 10815296 TI - Medication error prevention by pharmacists--an Israeli solution. AB - OBJECTIVE: The purpose of this study was to record prospectively the frequency of medication order errors in a general hospital in Israel with the objective of assessing the impact of pharmacist intervention in preventing potential harm. METHODS: The study was conducted during a 6-month period. A total of 160 medication order errors were detected at the hospital of which 60.6% were prescription errors and 39.4% were therapy ones. Principal types of errors detected were incorrect dosage (27.5%), interactions between drugs (20%), incorrect drug (12.5%), route (11.2%) and frequency (11.2%). Medication error rate by degree of severity was calculated per 100 patient days. The highest rate was found in Hemato-Oncology (2.48), followed by Intensive Care (0.82), Surgery (0.48) and Internal Medicine (0.26). Anti-infective drugs were the most prevalent class of drugs in which errors occurred (38.7%) followed by total parenteral nutrition preparations (21.8%), antineoplastics (15.6%) and anticoagulants (11.3%). Changes in medication orders due to pharmacists' intervention only occurred in 73.8% of error cases, most referring to dosage or route change (37.5%). CONCLUSIONS: This study underestimates the actual number of medication errors. However, it identified problem areas and trends so that better measures to improve drug use could be implemented. PMID- 10815297 TI - Erroneous intravenous administration of sustained action suspension of betamethasone. PMID- 10815298 TI - The need for clinical pharmacy. PMID- 10815299 TI - [Medicine and politics]. PMID- 10815300 TI - [Atherosclerosis: an infectious disease?]. PMID- 10815301 TI - [The difficult airway]. AB - Management of the difficult airway has gained increasing interest, because hypoxia is one of the leading causes of death and of severe neurological sequelae related to anesthesia or resuscitation. The difficult airway algorithm of the American Society of Anesthesiologists as well as the guidelines of the European Resuscitation Council provide recommendations for the prevention of difficulties in tracheal intubation and/or mask ventilation. Especially preoperative patient evaluation is of major importance. Patients history, oral and maxillofacial anatomy, pharyngeal and laryngeal structures as well as cervical spine mobility have to be assessed and awake fiberoptic intubation has to be performed in all cooperative patients with indices pointing towards difficult airways. If problems in intubating the trachea are encountered after induction of anesthesia and mask ventilation is adequate, one must call for help and decide rapidly whether to awaken the patient or to proceed with alternative intubation techniques (e.g. different laryngoscope blades, flexible fiberoptic scope or other fiberoptic techniques, lighted wand, retrograde intubation or surgical airway). In the potentially life-threatening "cannot intubate--cannot ventilate" situation either transtracheal jet ventilation, laryngeal mask airway, the esophageal-tracheal Combitube or a surgical airway have to be performed or have to be inserted immediately. These alternative methods have to be appropriately taught and--as far as possible--to be trained under routine conditions in order to master emergency situations. PMID- 10815302 TI - Serum homocyst(e)ine levels in women with preeclampsia. AB - BACKGROUND: Endothelial dysfunction has been described as the final common pathophysiological pathway in the development of preeclampsia. Since it has been suggested that homocyst(e)ine damages endothelial cells, we measured serum homocyst(e)ine levels in women with preeclampsia and in healthy pregnant women in order to find a new prognostic parameter for women with preeclampsia. METHODS: Forty-five women with preeclampsia and 45 healthy women with uncomplicated pregnancies, matched for age and parity, were entered into the study. Serum homocyst(e)ine levels were measured by gas chromatography-mass spectrometry analysis and correlated to clinical data. Logistic regression models were used to analyse the influence of serum homocyst(e)ine levels on the presence of preeclampsia versus healthy pregnant women and on the risk of premature termination of pregnancy due to preeclampsia. RESULTS: Median serum homocyst(e)ine levels in women with preeclampsia and healthy pregnant women were 14.2 (range 5.7-38.1) mumol/L and 15.1 (range 5.2-23.1) mumol/L, respectively (Mann-Whitney U-test, p = 0.8). In univariate logistic regression models, serum homocyst(e)ine levels had no significant influence on the odds of presenting with preeclampsia versus healthy pregnant women (univariate logistic regression model, p = 0.8) and on the odds of premature termination of pregnancy due to preeclampsia (univariate logistic regression model, p = 0.3). CONCLUSIONS: Serum homocyst(e)ine levels are not elevated in women with preeclampsia and are not associated with clinical outcome in women with preeclampsia. PMID- 10815303 TI - [Technique and results of laparoscopic conservative organ-preserving myoma enucleation]. AB - Myoma is one of the most common benign diseases of the female genital tract. The surgical management of this entity has been altered over the last years from complete hysterectomy to conservative enucleation of the myomas. We retrospectively compared our data concerning laparoscopic or open myomectomies. Over a period of 2 years, we operated 207 myomas in 102 patients. Of this collective, 69 (67.6%) were operated on laparoscopically and 33 (32.4%) via an open approach. In both groups, the median number of myomas was 2 (1-7). The mean diameter of the largest myoma was 5.1 +/- 2.4 cm (laparoscopy) and 6.2 +/- 2.6 cm (laparotomy), respectively. The additive diameter of myomas was 7.7 +/- 5.1 cm (laparoscopy) and 9.8 +/- 4.1 cm (laparotomy), respectively. There was no relevant difference between the groups in terms of operating time and blood loss. Four (3.9%) laparoscopies had to be converted to an open approach. In three cases (2.9%) a laparoscopically assisted enucleation had to be performed, requiring a mini-laparotomy 4 to 5 cm in length. We encountered no severe complications. Given appropriate indication, laparoscopic myomectomy is an easy-to-perform and minimally invasive technique with a low complication rate. PMID- 10815304 TI - [Daytime tiredness correlated with nocturnal respiratory and arousal variables in patients with sleep apnea: polysomnographic and EEG mapping studies]. AB - There is evidence that daytime tiredness is caused by apnea/hypopnea with oxygen desaturation and/or by sleep fragmentation due to arousals. The aim of this study was to investigate objective and subjective sleep and awakening quality and daytime vigilance--objectified by midmorning mapping of vigilance-controlled EEG (V-EEG)--in sleep apnea patients (N: 18), as compared with age- and sex-matched normal controls (N: 18) as well as to correlate nocturnal respiratory distress and arousals to daytime brain function. Statistical analyses demonstrated a deterioration in subjective and objective sleep and awakening quality in apnea patients. Midmorning V-EEG mapping in apnea patients exhibited less total power, more delta and theta, less alpha and beta activity, as well as a slower dominant frequency and centroid of the total activity compared to controls, which suggests a vigilance decrement. The Spearman rank correlation between 6 polysomnographically registered respiratory variables and 36 diurnal quantitative EEG measures demonstrated the following: the higher the apnea, apnea-hypopnea, snoring and desaturation indices and the lower the minimum and average low oxygen saturation, the more pronounced was diurnal tiredness. Eleven arousal measures based on ASDA criteria showed the following significant correlations: the higher the nocturnal arousal index and the more arousals due to hypopneas, the greater was daytime tiredness. On the other hand, the greater the average frequency change during arousals and the more spontaneous arousals, the better was daytime vigilance. Our findings show that, in contrast to the lengthy Multiple Sleep Latency (MSLT) and Maintenance of Wakefulness (MWT) tests which evaluate sleep pressure under resting conditions conducive to sleep, V-EEG mapping provides a brief objective measure of a sleep apnea patient's daytime tiredness under conditions of wakefulness more appropriate to reflect the patient's everyday life. PMID- 10815305 TI - Correlation between raised body temperature and acute mountain sickness score at moderate altitude. AB - In high altitude mountaineering, a rise in body temperature has long been associated with acute mountain sickness. No data exist on the situation at moderate altitudes in the Austrian Alps. The objective of this study was to investigate a potential relationship between an increase in body temperature and acute mountain sickness (AMS) and hypoxemia at moderate altitude. Body temperature and arterial oxygen saturation (SaO2) were measured in 40 alpinists at 1000 m altitude and after ascent to 3100 m altitude, and the AMS score was measured at 3100 m altitude. At 3100 m altitude, 3 alpinists with AMS (score 3) experienced a 0.87 +/- 0.12 degree C rise in body temperature and a 10.67 +/- 1.15% reduction in SaO2. In 8 moderately affected alpinists, temperature increased by 0.49 +/- 0.16 degree C and SaO2 was reduced by 6.75 +/- 1.75%. In 29 alpinists without signs of AMS, temperature did not change (difference 0.02 +/- 0.14 degree C) and SaO2 decreased by 4.59 +/- 0.82%. The difference between temperatures at the two altitudes correlated significantly with the SaO2 difference between the two altitudes (rs = 0.408, p < 0.01) and with the AMS scores (rs = 0.814, p < 0.01). CONCLUSION: Comparable with maximal forms of AMS at high altitude, our data provide reason to speculate that systemic inflammatory disease could also be causal in less severe forms of acute mountain sickness. Therefore, in cases of fever at moderate altitude, the differential diagnosis must include acute mountain sickness. PMID- 10815306 TI - Permissive hypoxemia permitted postoperative weaning from artificial ventilation after repair of pulmonary atresia. AB - Survival after corrective surgery of pulmonary atresia was associated with low right ventricular pressure, indicating normal pulmonary vascular resistance. Therefore increased fractional inspiratory oxygen concentration, inhaled nitric oxide and intravenous prostacyclin were considered to be effective measures during postoperative intensive care. In a 20-year-old female, conduit repair and unifocalisation of pulmonary atresia with ventricular septal defect and systemic to pulmonary arterial collaterals were performed despite preexisting one-sided pulmonary hypertension. During the following postoperative period, normal arterial oxygen saturation aimed at by means of a high fractional inspiratory oxygen concentration, resulted in persistent pulmonary oedema despite fluid restriction. After several trials of weaning from artificial ventilation, permissive hypoxemia was eventually successful. PMID- 10815307 TI - Effects of a silicone-coated polyamide net dressing and calcium alginate on the healing of split skin graft donor sites: a prospective randomised trial. AB - An open randomised prospectively controlled trial was performed to assess the healing efficacy, slippage rate and degree of discomfort on removal of calcium alginate and a silicone-coated polyamide net dressing on split skin graft donor sites. Sixteen patients were randomised to the calcium alginate group and 14 to the silicone-coated group. The donor sites were assessed at days 7, 10, 14 and up to day 21. The mean time to healing in the calcium alginate group was 8.75 +/- 0.78 days (range 7 to 14 days) compared to 12 +/- 0.62 days (range 7 to 16 days) for the silicone-coated group (p < 0.01). Although more silicone-coated dressings slipped (5 versus 1), the difference was not statistically significant. Pain during the first dressing change was assessed using a visual analogue pain scale. Although no significant differences were found between the groups, it was necessary to change the dressing protocol in the silicone-coated arm of the trial after entering the first two patients. Overlaid absorbent gauze adhered to the donor site through the fenestrations in the dressing necessitating the placement of paraffin gauze between the experimental dressing and the overlying cotton gauze. There was one infection in the study, occurring in the alginate group. Based on these results we recommend calcium alginate as the dressing of choice for split skin graft donor sites. PMID- 10815308 TI - Physiological differences for distinct somatic sensory modalities and sweating among the donor sites of cutaneous and fasciocutaneous free flaps. AB - Differences of sensation and sweating among the typical sites of cutaneous and fasciocutaneous flaps (scapular, lateral arm, radial forearm, groin and dorsalis pedis) were assessed in 30 healthy volunteers (20 males and 10 females) aged 17 62 years (mean 38.2 years). Standard clinical methods were used: Semmes-Weinstein monofilaments for testing light touch threshold, discriminator and blunt caliper for evaluation of static and dynamic two-point discrimination and the Marstock quantitative method for assessing the normative values of warm-cold difference limen and heat and cold pain thresholds. Spontaneous sweat secretion was observed and documented by the ninhydrin test. We established various physiological differences for distinct somatic sensory modalities and sweating among the body regions (donor sites of cutaneous and fasciocutaneous free flaps). PMID- 10815309 TI - Case report: a free arterialized venous flap for intraoral cancer reconstruction. AB - The free arterialized cephalic venous flap is a good option for intraoral reconstructions. It offers thin, pliable skin as a one-stage transfer without sacrifice of a major artery. The radial dominant hand poses a problem, which can be surmounted by this flap. PMID- 10815310 TI - CO2 and Nd:YAG laser-assisted nerve repair: a study of bonding strength and thermal damage. AB - To improve the laser welding procedure, a comparative study was undertaken to investigate the acute bonding strength and the thermal damage following CO2 and Nd:YAG laser-assisted nerve repair, performed with and without the use of blood and/or albumin as a solder. The strongest welds were produced with the CO2 laser using albumin as a solder. Thermal damage was minimal with the CO2 laser, whereas the damage with the Nd:YAG laser was substantial. The high bonding strength combined with minimal thermal damage of the nerve following repair with the CO2 laser with the use of albumin justify further investigations using this technique in in vivo studies. PMID- 10815311 TI - Cleft palate repair by Furlow double-reversing Z-plasty: first speech results at the age of 6 years. AB - The authors evaluated the first 15 six-year-old children operated on for cleft palate using double-reversing Z-plasty. Examination by a surgeon did not reveal a higher morbidity as a results of the new surgical method. Examination by a phoniatrist provided evidence of marked improvement of speech functions as compared with the previous von Langenbeck method. The results are preliminary, based on a small group of patients. PMID- 10815312 TI - Analysis of intracranial relations in patients with unilateral cleft lip and palate using cluster and factor analysis. AB - The present investigation was based on the cephalometric analysis of lateral teleroentgenographic pictures of 48 ten-year-old boys with complete unilateral clefts of the lip and palate. Using cluster and factor analysis, the author investigated the relationships among 75 craniofacial size, shape and position characteristics. Both multivariation methods proved useful in the search for associations between partial cranial structures. They supplemented each other well, and their combined use contributed to the definition of the following basic and specific principles of craniofacial relations and compensatory and adaptive mechanisms in patients with clefts. The maxilla consists of two mutually almost independent parts. The vertical parameters of the anterior upper face are interrelated with the slope of the palate plane; the dentoalveolar component of the upper jaw is formed independently. This fact can be aptly explained by the effect of orthodontic treatment, which contributes substantially to an improvement of the facial configuration. Rotation of the mandible has an impact on the vertical dimensions of the anterior lower, as well as the whole, face and is closely associated with the position of the mandibular body but not with the position of its branch nor with its length. The slope of the mandibular branch correlates with the mental angle, with the gonial angle and the slope of its dentoalveolar component, which is the result of adaptation to a shortening of the maxilla while preserving a positive overjet. Damage to the sagittal intermaxillary and interalveolar relations is primarily due to a reduction in the depth of the maxilla and is associated with the length of the mandibular body, while no relation with its rotation or length of the ramus was found. Solution of the given problem may help to find predictive models of facial development in patients with clefts. PMID- 10815313 TI - An integrated approach to communicable disease surveillance. PMID- 10815314 TI - Elimination of leprosy in the Americas. PMID- 10815315 TI - Argentina: health situation analysis and trends, 1986-1995. PMID- 10815316 TI - Case definitions. Neonatal tetanus. PMID- 10815317 TI - Case definitions. Tuberculosis. PMID- 10815318 TI - [The future of surgery]. PMID- 10815319 TI - [Enterovirus determination in cases with a diagnosis of the Guillain-Barre syndrome by using the acid-concentration technic]. AB - The purpose of this work was to detect enteroviruses in feces by an acid concentration technique (ACT). Fifty-eight samples from children less than 5 years age with diagnosis of Guillain-Barre syndrome (GBS) were analyzed using the routine technique and ACT. Nine positive samples with the routine technique were used as controls. Nine control samples and 22 additional (31 cases) non-polio enteroviruses were isolated and identified with the ACT (53%). Thus, 38% more isolates were obtained by ACT. Isolation was more successful in the RD cellular line (59%) than in Hep-2c (41%). In most cases most titers (71%) obtained were low. ACT improved the detection of enteroviruses but because it is very expensive and laborious, it should be used in the case of laboratories that analyze multiple samples, for special cases such as with autopsy cases and when results are compatible to poliovirus using the routine technique and only in samples obtained during the first 15 days of symptomatology. PMID- 10815320 TI - [Granulocyte colony-stimulating factor in the treatment of febrile neutropenia]. AB - PURPOSE: To determine whether granulocyte colony-stimulating factor (G-CSF) used in addition to antibiotic therapy, in patients with chemotherapy-induced febrile neutropenia shortens the period of fever, neutropenia and hospitalization. PATIENTS AND METHODS: The study was prospective. Patients with lymphoblastic acute leukemia (LAL) were included. They received intensive chemotherapy of induction, intensification, or consolidation. At random, a group received amikacin-ceftriaxone; if no had response after 3 days, we added vancomicin and, after 7 days, amphotericin. The other group received in addition these antibiotics, granulocyte colony-stimulating factor. RESULTS: The groups were comparable in the magnitude of the initial neutropenia (< 0.5 x 10(9)/L), site of the infection, chemotherapy received germs isolated, age, and sex. The patients of the group that received FEC-G were cured in the course of 3.1 days; in the group without FEC-G, this occurred in 7.2 days (p = 0.0001). At the end of the infectious episode, the number of neutrophils, in the group with FEC-G, was of 1.9 x 10(9)/L versus 0.7 x 10(9)/L (p = 0.0009). The mortality was of one and two cases (p = 0.46). The global mortality was 7.5%. CONCLUSIONS: The addition of FEC G to the treatment with antibiotics, in febrile neutropenia, decreases duration of days with fever, hospitalization and neutropenia. However, the frequency of cure is not augmented. PMID- 10815321 TI - [FARMAC: a program designed for monitoring the prescription of antimicrobials in hospitals]. AB - INTRODUCTION: The use of computer programs for monitoring the prescription of antimicrobial agents in hospitals is of paramount importance in the incorporation of electronic databases developed and analyzed by several hospital committees, including the following: infection control; pharmacy; morbidity and mortality; and quality assurance committees, as well as other key areas or departments such as pathology, pharmacy, microbiology, radiology, or hospital administration. OBJECTIVE: To describe the development and use of an "information system" employed for monitoring the prescription of antimicrobial agents in hospitalized patients. MATERIAL AND METHODS: A printed format was developed in order for physicians to ask for authorization for the use of restricted antimicrobial agents (i.e. third-generation cephalosporins and other newly introduced beta lactams, fluoroquinolones, antifungals, antivirals, etc). A computer program, "FARMAC", was designed using the Dbase IV program. FARMAC contemplates the presentation on a screen of a main menu allowing for the use of several options: admissions; antibiotic changes; consultations; discharges; hospital transfers; reports, and distribution. The reports are generated from the menu and appear on the screen or are printed in representative tables and figures. Generally speaking, the system processes information on the use of antimicrobial agents, files information, allows for consultation, of the information and generates reports quickly and efficiently. RESULTS: The presentations were defined on screen, taking into account the space needed in order to avoid an inadequate distribution of the messages, therefore obtaining an optimal image. A database was created defining the necessary fields for storing information with reference to patients and usage patterns of restricted antimicrobial agents. DISCUSSION: The development of an information and surveillance system on the prescription of antibiotics allows for consulting updated information, making the data analysis easier for decision making. Its use will allow for surveying tendencies and usage patterns of controlled and non-controlled antimicrobial agents. PMID- 10815322 TI - [Dynamic pancreatography: the diagnosis of pancreatic necrosis and the identification of the risk factors of severity]. AB - BACKGROUND: Dynamic pancreatography (DP) can establish the development of tissue damage and demonstrate pancreatic necrosis (PN). OBJECTIVE: To prospectively evaluate the usefulness of DP in patients with acute pancreatitis (AP), analyzing its capabilities for determining the incidence of necrosis, and as a prognostic predictor. MATERIAL AND METHODS: All patients with AP seen at our department during 4 years were included. All patients were scheduled to DP. Findings were classified in five groups: I, interstitial AP; II, secondary fluid collections; III, peripancreatic necrosis; IV, pancreatic necrosis < 50% and pancreatic necrosis > 50%. Also included for analysis were: demographic variables, etiology, APACHE-II score, and Ranson's signs; when infection was suspected, cultures were done. PD and operative findings were correlated to calculate sensibility (S), specificity (Sp), positive and negative predictive values (PPV, NPV), and the diagnostic accuracy (DA) of the test. Correlation between severity/mortality and morphologic alterations according to the classified group were also performed. RESULTS: 244 patients. Biliary associated pancreatitis in 47% and alcohol induced in 37%. DP in 207 patients showed, 34 patients in group I; 36 in group II, 67 in group III, 47 and 60 in groups IV and V, respectively. Secondary pancreatic infection was present in 48%. One hundred seventy-two patients underwent surgery; the surgical indication was to interrupt the pathogenesis of complications in 52%. DP correlated with the prognostic signs and APACHE-II score as predictors of complications with an overall (S) of 93%, (Sp) 98%, (PPV) 91%, (NPV) 98%, and (DA) 97%. Greater scores values are all directly proportional with PD necrosis findings (P < 0.05). There was not PD related morbidity. CONCLUSIONS: We found correlation between PD classification and prognostic factors. Morphologic alterations were accurately identified. PD as preoperative work-up allows surgeons design operative strategies and as is useful follow-up of the patients after necrosectomy. PMID- 10815323 TI - [Cushing syndrome. The diagnostic and therapeutic challenges]. AB - Endogenous hypercortisolism is characteristically a condition that should be diagnosed and treated in tertiary care centers with the participation of several specialists. Cushing's syndrome represents the clinical expression of a prolonged exposure to glucocorticoids, independently of its origin. The term Cushing's disease refers to the hypercortisolism that results from the excessive secretion of corticotropin (ACTH) by a pituitary microadenoma. The mechanisms that give rise to the different forms of hypercortisolism are complex and its precise differential diagnosis is one of the major challenges in modern endocrinology. This review focuses on current aspects of the pathophysiology, differential diagnosis and treatment of Cushing's syndrome. PMID- 10815324 TI - [The psychodynamics of deafness]. AB - OBJECTIVE: Deafness generates psychological conflicts for the deaf as well as for others in their environment. Our purpose is to obtain a panorama of the topic based on the available bibliography. INFORMATION: Relevant articles were located in Medline and in unlisted journals, accessible at the National Institute of the Human Communication and at the Juan N. Navarro Hospital of Mexico City. The books were located at the National Institute of the Human Communication. SELECTION OF THE MATERIAL: The articles were selected by the authors, based in their apparent internal consistency and relation to our purpose. Two different concepts became apparent: one which recognizes a "personality of the deaf" that would predispose to emotional damage, and the other that denies that concept, explaining the distinctive features of deaf people as due to basically environmental factors. CONCLUSION: We think that this review sustains the premise that the psychological damages of deafness are real, not inherent to it, but caused by the social, occupational, pedagogical and familiar conflicts that are generated. Those damages vary with the age at the presentation of deafness the personal adjustment to the deficiency and acceptance by the community. The mitigation of deafness, damage depends on educated parents, adequate instruction programs (scholastic as well as occupational), and when needed, appropriate supportive psychotherapy based on the specific needs of each deaf person and this/her environment. We consider highly desirable to promote the elaboration and publication of interdisciplinary studies related to this topic. PMID- 10815325 TI - [A 43-year-old woman with a paraneoplastic syndrome]. PMID- 10815326 TI - [Malfunction of a permanent pacemaker]. AB - Forty years have gone by since the first pacemaker implant; this fact had strong impact in the life of thousands of persons. The objective of this work is to report the case of definitive pacemaker malfunction with epicardiac lead and review the literature concerning the important aspects of the causes and diagnosis of pacemaker malfunction. We consider that physicians dealing with patients implanted these devices must be prepared to diagnose and treat them adequately. PMID- 10815327 TI - [Historical notes on the beginning of experimental surgery in Mexico (1918 1936)]. AB - This is a historical review from 1918 to 1936 concerning the early papers written in Mexico on experimental surgery. From them, we can learn how Mexican surgeons attempted to repeat the results of authors U. S. and Europe. Additionally, the problem of not having good statistical information in Mexico is illustrated. These papers were published in Mexican journals of limited circulation or in graduation theses. The most important works were ureteroplasties repair in humans with canine tissues (F. Quiroz, 1919). Physical aspects of abdominal sepsis (C. Robles, 1929), and bile duct prosthesis, (Gomez Guerra, 1932). This paper pays homage to Mexican surgeons who in spite of enduring financial difficulties and times of political turmoil, achieved the beginning of these areas of surgery in Mexico. PMID- 10815328 TI - [The transmission of typhus by the louse: Dr. Miguel Otero (1906)]. PMID- 10815329 TI - [The first separation of Siamese twin sisters: Dr. Aureliano Urrutia 1917]. PMID- 10815330 TI - [Is it possible to create human malignant cells in the laboratory?]. PMID- 10815331 TI - [An update on the pharmacological therapy of the andropause]. PMID- 10815333 TI - [In Process Citation] PMID- 10815332 TI - [Unnecessary operations in the exercise of surgery. A topic of our times with serious implications in medical ethics]. AB - Unnecessary operations represent between 20 and 25% of total surgical practice. This article analyses causes, history, enormous economic effects and their consequences as an iatrogenic harm. Looking for a solution to the problem of unnecessary operations is urgent because these surgeries have a direct effect on the field of ethics and morality in the practice of surgery. PMID- 10815334 TI - [The contribution of social medicine to public health]. PMID- 10815335 TI - [Awarding the 1999 Salomon-Neumann Medal to Prof. Dr. rer. biol. hum. Hans-Konrad Selbman]. PMID- 10815336 TI - [Social medicine components of quality management and what social medicine can learn from quality management. Address at the presentation of the 1999 Solomon Neumann Medal of the German Society of Social Medicine and Prevention]. PMID- 10815337 TI - ["Managed care"--perspectives in Germany]. AB - There is a public discussion in Politics and Health services whether managed care should be realized in Germany. The German Medical Services of the Statutory Health Insurance (MDK) are also involved and have great interest and an opinion about this topic because the introduction of managed care or elements of managed care will influence the cooperation between the MDK and the German Statutory Health Insurance. The following article informs on certain elements of managed care already practised in Germany and discusses which of these elements can be used more intensively in the future according to Social Laws and developed structures. PMID- 10815338 TI - [Physician's anxiety and physician's elegance. Problems in dealing with cost reduction, education of general practitioners and optimal size of practice networks in a cross-national comparison]. AB - The key reason for physicians networking in managed care is to get a better coping with uncertainty on action (treatment) decisions. The second reason for networking in managed care are financial benefits grounds. But this reason is very ambivalent. Three different action problems (role conflicts) in managed care network are to solved, which was also in single practices. In the lecture the decision strategies and decision resources has been compared. Observations are done using expert interviews, patient interviews and analysis of documents in USA, Germany and Switzerland. The first problem is the choosing of a cost reduction strategy which is not reducing the effectiveness. Such "ugly" solution strategies like exclusion of "expensive" patients and a rationing of necessary medical services in a kind of McDonalds network of physicians will fail the target. The optimost way is a saving of all unnecessary medical even injourious performances. The chosen cost reduction strategy is not real visible from outside but in fact limited cognizable and controllable. Evidence based health care can be a resource of treatment decisions and could train such decisions but it will not substitute these decisions. The second problem is the making of real family practitioners as gatekeepers. Knowledge about the care system is still not making a real family practitioner, even if this is the minimum condition of their work. Also contractual relationships between insurance and doctor as a gatekeeper or financial incentives for patients are still making not a real family practitioner as a gatekpeeper. Only throughout the trust of patients supported by second opinions is making the real family practitioner as a gatekeeper. "Doctor hopping" could be the reaction by scarcity of trustworthy family practitioners as gatekeepers. The third problem is the choosing of the optimal scale of a network due to the very different optimal size of networks regarding the requirement of risk spreeds, of the motivated engagement, of competition, incentives of inclusion of insurantes, they always need other net sizes. But it is possible, for each requirement there could function different networks. A practice (doctor's office) can be a member in different networks in several levels. The social transition from a small office to a network of offices is in all business lines a cultural shock involving not only benefits also psychical and social distress. In this there is no difference between health or agriculture or each other business of trade and industry. The destiny of the joint doctor's offices in Germany suggest due to a very serious power to scatter this networks. The comparative analysis of conflicts, strains, resources and strategies of associations and networks could yield from a developed methodical repository in sociology and social psychology what exists since 40 years (see also Meyer--in this journal). But therefore must be included also the action problems, which are only mentioned in passing of the according profession horizon. PMID- 10815339 TI - [Social change in the physician's role and medical practice caused by managed care in Switzerland]. AB - Switzerland is the first European country where health maintenance organizations (HMOs) characterised by capitation (per capita lumpsum) and gatekeeping were implemented according to the HMO staff model known in the USA. The development of managed health care in Switzerland relies on the belief that adequate economic incentives and competition result in cost reduction and high quality health care. Whether this is true or not--in any case the deregulation of legally accepted forms of health insurance and managed care result in profound changes in the Swiss health care system. Observations are made by using expert interviews and analysis of documents. The implementation of managed care induces socio-cultural changes of the medical profession which are as profound as the induced economic changes. We discuss conflicts of interests among physicians using four main dimensions of conflict: (1) control, (2) monopolization, (3) valuation, and (4) specialization. In the HMOs we observe pronounced conflicts of the physicians' role. The changes of the physicians' role in HMOs is on the one hand the result of new duties. On the other hand it expresses strategies of coping with the role conflict between the main clinical duties and the new obligation to control cost and to monitor treatment via gatekeeping. In HMOs the teamwork of doctors and the quality control of care promotes the satisfaction of physicians with their work, however, it can also have dysfunctional effects. PMID- 10815340 TI - [Continuing education, motivation for cost control, a Swiss experiment]. AB - The article reviews the prospective, controlled experience of the introduction of a physician network service (health maintenance organisation 5000 insured persons). There was a reduction of cost (26%-33%) compared with the control group, without loss of quality. This reduction still holds good after four years. Risk selection explains only a small part of the reduction in cost. The only interventions consisted of gatekeeping plus continuing medical education (CME). Diagnosis and financial data were not available and profiling was not used to control the experience. CME was provided by a quality circle once a week. In these meetings, the subjective and objective features particular to outpatient medicine were addressed. PMID- 10815341 TI - [Subjective health and health behavior of children and adolescents--a survey of Hamburg students within the scope of school medical examination]. AB - The subjectively perceived health of children and adolescents has received little attention in Public Health research so far. In co-operation with school physicians of Hamburg, Eimsbuettel, 1,501 children and adolescents (grades 4 and 8, as well as their parents) were questioned with regard to health-related quality of life and health behaviour, using among others the short form of the KINDL Quality of Life Questionnaire and established psychosocial instruments. High values in the KINDL scale were found in 75% of the students, the mean value of the 0-100 transformed KINDL total scale was around 80. Adolescents--and in this group specifically the girls--reported significantly lower quality of life scores. Psychosocial strain, presence of disease and school absenteeism were negatively associated with quality of life ratings. Health-related behaviours in terms of resources in the area of food intake and risk behaviours in terms of addiction, obesity and stressors correlated with subjective health as expected. In multiple regressions these factors explained 20% of the KINDL-score variance in the total group. Taking into account indicators of functional status and life satisfaction, the percentage of explained variance increased to over 50%; here these predictors yielded a correct classification of high vs. low self-reported quality of life in 90% of the cases. The study shows that subjective health is a methodologically adequate and scientifically interesting indicator which in the public health area can be used in epidemiological research, in health reporting and in school physicians' practice to identify young persons with quality of life impairments in order to provide early and in-focus support to this group. PMID- 10815342 TI - [Concept for a health care economic evaluation of short- and long-term costs and effectiveness parameters of an expanded ambulatory psychosomatic rehabilitation program]. AB - There are only few ambulatory rehabilitation concepts for mothers with psychosomatic disorders (prevalence 5%). Also, only little is known about the evaluation of these programmes. This study compares the socioeconomic evaluations of an ambulatory rehabilitation programme with a post-assistance programme and one without a post-assistance programme. The superior programme should be determined by weighing all relevant costs and benefits. The intensive phase of the ambulatory rehabilitation programme consists of an eight-week treatment with group, single, body and art therapy. The post-assistance programme spans a period of nine-months with 36 sessions of psychoanalytic group therapy. The evaluation of both programme alternatives is made by with the evaluation tool of socioeconomic analyses. For consideration and pricing of costs all direct medical costs, direct non-medical costs and indirect costs are being monitored. Outcomes assessment is realised by cost-effectiveness analysis, cost-utility analysis and cost-benefit analysis. Utilisation of the health care system is being assessed with questionnaires. Measurements are being performed at the beginning and the end of the rehabilitation programme and three, six, nine and twelve months later. Mothers with children aged six years and younger and suffering from various psychosomatic disorders were included in this study. The costs identified for the rehabilitation programme are DM 5571.10 (intensive care and post-assistance programme) and DM 1512.40 (intensive care) per patient. Further progress of the study will show if future cost will differ between the two alternatives. For the comparison of both alternatives all costs will be linked with outcomes. It remains to be seen that additional costs of the post-assistance programme will be compensated with positive outcomes. A sensitivity analysis will show if variation of assumptions will influence the cost-benefit-ratio of the different alternatives. PMID- 10815343 TI - [An integrated concept for contraception counseling and HIV prevention for young women: the Project Necessaire]. AB - PURPOSE: The aim of this study was to evaluate the effectiveness of the innovative project "Necessaire" on sexual and reproductive health promotion. "Necessaire" was developed for young women and carried out in the German-speaking part of Switzerland. The concept consisted of giving theoretical information in a brochure suitable for young women in its design combined with several items (such as condom, chewing gum, sticker) in a spongebag. METHOD: Young women (n = 183) as well as professionals (n = 154) were asked to give their opinion on the "Necessaire" by filling in a semi-structured questionnaire. In addition, the product ordering was monitored over 7 months, in order to examine the use of the product by various institutions. RESULTS: On the whole, the young women praised the product. They especially liked the brochure, which was made up like a diary written by a young woman called Rosa. According to their answers the combination of information and useful items stimulated the reflection on topics like sexuality, love, and prevention of sexually transmitted diseases. Critical voices pointed out the need for interventions aimed at young men, in order to make them more aware of their responsibility in contraception, HIV prevention and partnerships. The professionals generally would like to continue using the product in their work with young women. Many of them asked for more accompanying material with practical ideas for using the product. A similar product for young men and foreign youngsters was often asked for. The monitoring of the product ordering showed that schools and institutions such as youth centres evinced the greatest interest in the product. This contrasted with the results of a feasibility study conducted in advance. Although medical and advice centres indicated a need for a product to support professionals counselling young women, these institutions ordered relatively few kits. CONCLUSION: Generally speaking the "Necessaire" was well received by young women as well as by professionals. It not only informs on sexual health but also promotes preventive behaviour in daily life. The product was not used as an aid for counselling young women as much as was intended. It is suggested to promote the Necessaire's use by taking specific measures like developing written material with practical ideas for professionals. In the case of developing a similar product for young men and migrants, their specific backgrounds and cultures should be taken into account and the material adapted accordingly. PMID- 10815344 TI - [Who is rehospitalized in a psychiatric hospital? Psychiatric hospitalization rates and social indicators in the Zurich canton (Switzerland)]. AB - There are two approaches in the research on the relation between social conditions and mental disorder: The ecological approach is concerned with characteristics of the social composition of a certain geographical area and their relation to the frequency of disorders, whereas for the individualistic view variables of the psychosocial background of the individual are of interest. This study is on the risk for psychiatric admission (first and re-admission). While considering variables of the social context of the community as well as of the background of the individual, it tries to take into account both the ecological and the individualistic view of the relationship between social conditions and (treated) mental disorder. The sample of the study includes data of 4021 psychiatric inpatients treated in 1997 in one of the seven psychiatric hospitals of the Swiss canton of Zurich as well as data of social context of the 171 communities of the canton of Zurich. The psychiatric first and re-admission rates of the community can be predicted by the following variables of its social context: 1. pro portion of foreigners, 2. urban character of the living area, 3. population density. Two other variables are of relevance only for the prediction of first admissions: 4. proportion of one-person households and 5. local tax rate. However, further results of the study show that correlations between variables of the social context and psychiatric admission rate of the community cannot be interpreted as risks for the individual. PMID- 10815345 TI - [An instrument for evaluating the psychiatric personnel regulation]. AB - Since 1991, the Psychiatrie-Personalverordnung (Psych-PV) regulates the allocation of personnel in German psychiatric hospitals and psychiatric wards. Psych-PV defines the personnel's diagnostic and therapeutic tasks based on an illness severity scale. A group of experts from the Medical Advisory Services of Social Security--Health Insurance (MDK) has developed a questionnaire as an instrument to systematically gather, from a given hospital, structural data relevant for application of Psych-PV. The questionnaire consists of two parts, relating to the hospital and to the individual wards. The hospital supplies information such as statutory catchment area, types of care offered, capacity, personnel, organisational structure, patients' diagnoses, and co-operation with complementary institutions. The section pertaining to individual wards collects data on admission and discharge, duration of treatment, re-admissions, and internal transfer. The distribution of the severity of patients' illnesses within individual wards, which has been defined in the budget, is the basis for a review of the actual number of staff as well as of the realization of basic tasks of diagnostics and treatment. Questions pertaining to treatment concepts in individual wards are aimed to ascertain the degree of agreement with the conceptual goals of Psych-PV. The methods of treatment offered in a given ward can be gleaned from its weekly organisation-plan. Indications for the realization of basic tasks of diagnostics and treatment can be derived from the information supplied by each personnel group of a ward with regard to specific tasks of individual and group treatment. The questionnaire developed by the MDK in Hessen allows to review the extent to which the allocation of staff according to Psych PV on the level of the individual ward translates into appropriate treatment options. The questionnaire also offers a basis for a comparison of hospitals. PMID- 10815346 TI - [Prevalence and risk factors of iron deficiency in young mothers]. AB - Quality of life and achievements are impaired by unrecognised iron deficiency. The iron requirement of women during their child-bearing age is high and increases in pregnancy. The aim of this study was to determine the prevalence and risk factors for iron deficiency in young mothers under contemporary German life conditions. Between September 1997 and August 1998 the iron status of 507 mothers of one-year old children was assessed. The data was derived from venous blood and questionnaires. Besides conventional methods, the concentration of soluble transferrin receptor was used as leading indicator of iron status. 9.5% had cellular iron deficiency and 2.2% of all mothers had iron deficiency anemia. In addition to absence of school education non-German nationality, a high number of children and vegetarian food are risk factors for iron deficiency. In contrast, high alcohol intake and cigarette smoking are associated with a better iron status. Children of mothers with insufficient iron supply are also at higher risk of iron deficiency. PMID- 10815348 TI - 37th Congress of the Italian Society of Hematology. Torino, Italy. September 26 29, 1999. Abstracts. PMID- 10815347 TI - [Foundations--a means for implementing strategic goals and measures in public health]. AB - Social innovations are happening in many critical areas. Foundations make an enduring contribution towards increasing access to innovations in public welfare based on the philosophy that believes state-run organizations are neither efficient nor responsive to the people's changing needs. In this sense, foundations help to close the gap by turning hitherto tolerated conditions into problems and claims to action. The effectiveness of voluntary bodies as advocates of change owes much to their informal nature. In Germany, voluntary actions in Public Health are still underrepresented. Therefore, donors should be more supported by state government through adequate regulations. In addition, funders need to concentrate their efforts on the limited number of areas where they can have the greatest impact. PMID- 10815349 TI - Implementing the findings of health technology assessments. If the CAT got out of the bag, can the TAIL wag the dog? AB - OBJECTIVE: To explore whether more could be done to increase the implementation of health technology assessment (HTA) findings. METHODS: A literature review was undertaken to identify the main barriers to implementation, the mechanisms that influence the diffusion and use of health technologies, and evidence of the successful implementation of findings. RESULTS: Numerous barriers to the implementation of HTA findings exist at the public policy, healthcare professional, and general public levels. Several mechanisms for influencing the use of health technologies exist, and there are some examples of findings being implemented through these mechanisms. However, there are also concerns about the aggressive implementation of findings. A balanced approach to the implementation of HTAs is required. CONCLUSION: The main elements of a successful implementation strategy are: a) defining a clear policy question; b) defining a clear research question; c) making recommendations commensurate with the evidence; d) identifying the implementation mechanism; e) paying attention to incentives and disincentives; and f) clarifying the roles and responsibilities of the various parties. Further research is also required into several aspects of implementation. PMID- 10815350 TI - Evaluation of randomized controlled trials on complementary and alternative medicine. AB - OBJECTIVES: Use of complementary and alternative medicine (CAM) is growing in all Western countries. The goal of this study was to evaluate quality of randomized controlled trials (RCTs) of CAM interventions for specific diagnoses to inform clinical decision making. METHODS: MEDLINE and related databases were searched for CAM RCTs. Visual review was done of bibliographies, meta-analyses, and CAM journals. Inclusion criteria for review and scoring were blinded RCT, specified diagnosis and intervention, complete study published between January 1, 1966 and July 31, 1998 in an English-language, peer-reviewed journal. Two reviewers independently scored each study. RESULTS: More than 5,000 trials were found, but only 258 met all study inclusion criteria. The main cause for rejection (> 90%) was that the study was not an RCT or had no blinding. Mean score across 95 diagnosis/intervention categories was 44.7 (S.D. +/- 14.3) on a 100-point scale. Ordinary least-squares regression found date of publication, biostatistician as author or consultant, published in one of five widely read English-language medical journals and diagnosis/intervention category of hypertension/relaxation as significant predictors of higher scores. CONCLUSIONS: The overall quality of evidence for CAM RCTs is poor but improving slowly over time, about the same as that of biomedicine. Thus, most services are provided without good evidence of benefit. PMID- 10815351 TI - Cost-effectiveness of targeted screening for abdominal aortic aneurysm. Monte Carlo-based estimates. AB - OBJECTIVES: This article reports a cost-effectiveness analysis of targeted screening for abdominal aortic aneurysm (AAA). A major emphasis was on the estimation of distributions of costs and effectiveness. METHODS: We performed a Monte Carlo simulation using C programming language in a PC environment. Data on survival and costs, and a majority of screening probabilities, were from our own empirical studies. Natural history data were based on the literature. RESULTS: Each screened male gained 0.07 life-years at an incremental cost of FIM 3,300. The expected values differed from zero very significantly. For females, expected gains were 0.02 life-years at an incremental cost of FIM 1,100, which was not statistically significant. Cost-effectiveness ratios and their 95% confidence intervals were FIM 48,000 (27,000-121,000) and 54,000 (22,000-infinity) for males and females, respectively. Sensitivity analysis revealed that the results for males were stable. Individual variation in life-year gains was high. CONCLUSIONS: Males seemed to benefit from targeted AAA screening, and the results were stable. As far as the cost-effectiveness ratio is considered acceptable, screening for males seemed to be justified. However, our assumptions about growth and rupture behavior of AAAs might be improved with further clinical and epidemiological studies. As a point estimate, females benefited in a similar manner, but the results were not statistically significant. The evidence of this study did not justify screening of females. PMID- 10815352 TI - Patient, physician, and payer perceptions and misperceptions of willingness to pay for diagnostic certainty. AB - Little is known about the value patients, physicians, and payers place on intangible attributes of care. Differences in valuations among these groups and misperceptions of value of intangible attributes to other groups can contribute to conflicts about treatment recommendations or coverage decisions. We surveyed patients, physicians, and managed care executives to assess their willingness to pay (WTP) for diagnostic certainty for peptic ulcer disease (PUD) and gastroesophageal reflux disease (GERD). To determine if patients, physicians, and payers accurately perceive each other's valuations of diagnostic certainty, participants were also asked to estimate the WTP of each of the other types of respondents. Patients were most likely, and executives least likely, to value diagnostic certainty. For PUD, 84% of patients, 61% of physicians, and 43% of executives expressed a positive WTP. Median WTP was low for all three groups ($1 9 for patients and physicians; $0 for payers). Physicians and executives both correctly predicted patient WTP. For GERD, 87% of patients, 52% of physicians, and 29% of executives expressed a positive WTP. Executives underestimated patient WTP. For both diseases, physicians' WTP was overestimated by patients and underestimated by executives. The inconsistency in the value that patients, physicians, and managed care executives place on diagnostic certainty indicates the potential for conflict over practice guidelines or access to services. WTP surveys can provide information to aid in anticipating and addressing areas of disagreement. PMID- 10815353 TI - Willingness to pay for obesity treatment. AB - OBJECTIVES: To estimate willingness to pay (WTP) for effective treatment in a sample of obese individuals and to examine whether WTP is associated with factors reflecting the severity of obesity as well as a number of other variables such as age, sex, education, and income. METHODS: WTP and data on the severity of obesity were collected from the study, Swedish Obese Subjects. Associations between WTP, income, and obesity-related factors were analyzed by linear regression. RESULTS: The mean age was 47 years (range, 37 through 59 years) and the mean body mass index (BMI) was 39.6 kg/m2 (n = 3,549). Average personal monthly income was SEK 13,000 (approximately US $1,585), average WTP was SEK 26,900 (approximately US $3,280) and the median value was SEK 10,900 (approximately US $1,330). A high WTP was associated with high personal and household income, high weight, high education, female sex, poor perceived health, low current age, and low age at onset of obesity. Over 50% of the patients deemed it necessary to borrow money to cover their WTP. When adding a loan to the regression analysis, the associations between WTP and perceived health, age, and gender disappeared. CONCLUSION: Obese patients are willing to pay approximately twice their monthly salary for effective treatment and a higher WTP is associated with higher weight and poorer perceived health. PMID- 10815354 TI - Willingness to pay for arthritis symptom alleviation. Comparison of closed-ended questions with and without follow-up. AB - OBJECTIVE: To compare two methods of measuring willingness to pay (WTP): closed ended questions with and without follow-up. METHODS: A measurement experiment based on dichotomous choice contingent valuation survey data is reported. Marginal WTP estimates for alleviation of rheumatoid arthritis symptoms resulting from treatment with a novel anti-rheumatic agent, cA2 (TNF-alpha blockade), were calculated. Monte Carlo simulations were undertaken to evaluate the methods with respect to their statistical power. RESULTS: The estimated marginal WTP values using closed-ended questions with and without follow-up were DKK 637 (US $91) and DKK 1,268 (US $181), respectively. A Wilcoxon's signed-rank test showed that the difference of DKK 631 was significant. Moreover, including a follow-up question increases the precision of the result. Monte Carlo simulations showed that trade offs between power (i.e., the probability of a correct rejection of a false null hypothesis), efficiency, and size may exist in the two models. CONCLUSIONS: There was a significant difference between the WTP estimates when using closed-ended questions with and without follow-up. When choosing between the models, however, power, efficiency, and size could be used as selection criteria. PMID- 10815355 TI - Prioritizing investments in health technology assessment. Can we assess potential value for money? AB - OBJECTIVE: The objective was to develop an economic prioritization model to assist those involved in the selection and prioritization of health technology assessment topics and commissioning of HTA projects. METHODS: The model used decision analytic techniques to estimate the expected costs and benefits of the health care interventions that were the focus of the HTA question(s) considered by the NHS Health Technology Assessment Programme in England. Initial estimation of the value for money of HTA was conducted for several topics considered in 1997 and 1998. RESULTS: The results indicate that, using information routinely available in the literature and from the vignettes, it was not possible to estimate the absolute value of HTA with any certainty for this stage of the prioritization process. Overall, the results were uncertain for 65% of the HTA questions or topics analyzed. The relative costs of the interventions or technologies compared to existing costs of care and likely levels of utilization were critical factors in most of the analyses. The probability that the technology was effective with the HTA and the impact of the HTA on utilization rates were also key determinants of expected costs and benefits. CONCLUSIONS: The main conclusion was that it is feasible to conduct ex ante assessments of the value for money of HTA for specific topics. However, substantial work is required to ensure that the methods used are valid, reliable, consistent, and an efficient use of valuable research time. PMID- 10815356 TI - Prioritization attitudes among doctors and nurses examined by a scenario method. AB - OBJECTIVES: To investigate doctors' and nurses' attitudes toward 14 potential prioritization criteria. METHODS: This study was performed by using the random paired scenario method. The respondents received a questionnaire with 12 pairs of scenarios, imaginary patient cases, each of which contained two to three different prioritization criteria (e.g., child, old patient, poor patient). Respondents were asked which one of each scenario pair they would choose if only one patient could be treated. The scenarios were randomly put into 30 different questionnaire sets. There was a random selection of 241 doctors and 151 nurses in Finland, with response rates of 60.3% and 50.3%, respectively. RESULTS: Doctors prioritized young patients, severe disease, expensive treatments and posteriorized (negatively prioritized) demented or institutionalized patients, and patients having a self-caused disease. Children were strongly prioritized, even over serious diseases. Expensive treatments appeared to be favored by doctors, and this result cannot be explained by severity of disease. Nurses' attitudes were similar to those of doctors. CONCLUSIONS: Children were strongly prioritized. Elderly persons were posteriorized if they had dementia or were living in institutions. Patients having a self-caused disease are posteriorized, more often by nurses than by doctors. PMID- 10815357 TI - A tool to improve quality of reporting published economic analyses. AB - OBJECTIVES: To test the feasibility of obtaining a baseline level of quality of reporting for cost-utility analysis (CUA) studies using the British Medical Journal economic submissions checklist, test interrater reliability of this tool, and discuss its longer term implications. METHODS: CUA studies in peer-reviewed English language journals in 1996, assessed using the British Medical Journal checklist, a quality index, and interrater reliability correlations. RESULTS: Forty-three CUA studies were assessed, with 23 checklist items acceptable and 10 items inadequate. Lowest quality scores were reported in specialist medical journals. Proportional agreement between assessors was over 80%. CONCLUSIONS: The British Medical Journal checklist is a feasible tool to collect baseline information on the quality of reporting in journals other than the British Medical Journal. Editors of specialist medical journals are in greatest need of economic guidance. If handled carefully, they might consider adopting the British Medical Journal checklist. PMID- 10815358 TI - Measuring costs in cost-utility analyses. Variations in the literature. AB - OBJECTIVES: Although cost-utility analysis (CUA) has been recommended by some experts as the preferred technique for economic evaluation, there is controversy regarding what costs should be included and how they should be measured. The purpose of this study was to: a) identify the cost components that have been included in published CUAs; b) catalogue the sources of valuation used; c) examine the methods employed for estimating costs; and d) explore whether methods have changed over time. METHODS: We conducted a comprehensive search of the published literature and systematically collected data on the cost estimation of CUAs. We audited the cost estimates in 228 CUAs. RESULTS: In most studies (99%), analysts included some direct healthcare costs. However, the inclusion of direct non-healthcare and time costs (17%) was generally lacking, as was productivity costs (8%). Only 6% of studies considered future costs in added life-years. In general, we found little evidence of change in methods over time. The most frequently used source for valuation of healthcare services was published estimates (73%). Few studies obtained utilization data from RCTs (10%) or relied on other primary data (23%). About two-thirds of studies conducted sensitivity analyses on cost estimates. CONCLUSIONS: We found wide variations in the estimation of costs in published CUAs. The study underscores the need for more uniformity and transparency in the field, and continued vigilance over cost estimates in CUAs on the part of analysts, reviewers, and journal editors. PMID- 10815359 TI - A comparison of three approaches for attributing hospitalizations to specific diseases in cost analyses. AB - OBJECTIVES: Calculations of healthcare costs rarely disclose the specific approach used to allocate the cost of hospitalizations by diagnosis. However, the type of approach used can have a major impact on the findings in the case of significant comorbidities. The present analyses compared three approaches for attributing Medicare DRG reimbursements (which were used as surrogates for average costs) for hospitalization by diagnosis. METHODS: Medical resource utilization data from the National Hospital Discharge Survey were analyzed using each of three allocation approaches: a) attributing 100% of the cost of hospitalization to the disease when it was the first-listed diagnosis; b) attributing a portion of the cost of hospitalization to the disease, depending on its position in the list of diagnoses and the relevance of any comorbidities; and c) an incremental analysis of cost based upon the hospitalization experiences of an age and gender matched cohort. These three approaches were applied to the cost of hospitalization for chronic obstructive pulmonary disease (COPD). RESULTS: The first approach projected 206,098 hospitalizations at $3,449 per hospitalization for a projected U.S. annual total of $711 million. The second approach projected 681,547 hospitalizations at $3,205 per hospitalization for a projected U.S. annual total of $2.2 billion. The third approach also projected 681,547 hospitalizations, but at $2,361 per hospitalization, for a projected U.S. annual total of $1.6 billion. CONCLUSIONS: Expanding from the example on COPD, the limitations of each approach are described and their applications to other conditions are presented. PMID- 10815360 TI - A comparison of standard gamble, time trade-off, and adjusted time trade-off scores. AB - OBJECTIVE: To adjust patients' time trade-off (TTO) scores using information on their utility functions for survival time to derive a measure of health state utility equivalent to the standard gamble (SG). METHODS: A sample of 199 cardiovascular patients were asked three TTO and SG questions (to assess their own health state), and three certainty equivalent questions (to assess their utility function for survival time) in an interview. RESULTS: Patient's utility functions for time were increasingly concave, but being unable to model this successfully, a constant function with an averaged level of concavity was used. The raw TTO scores were significantly higher than SG scores, while the adjusted TTO scores were equivalent to the SG. CONCLUSIONS: Raw time trade-off scores will give biased estimates of health state utility when patients' utility functions for time are not linear, but these can be adjusted to yield true utilities. The constant proportional risk-posture assumption of the conventional QALY model, on which previous attempts to adjust time trade-offs have been based, was not supported by the data. PMID- 10815361 TI - A cost-benefit analysis of mechanical ventilation. An examination of DRG 475. AB - OBJECTIVE: An economic evaluation of the resources used for mechanically ventilated patients using various measures for the benefits of extending life. METHODS: Regression analysis is applied to New York State discharge data for patients under DRG 475 during 1992-96 to predict age-specific survival rates and payments per life saved. Sensitivity analysis is used to compare benefits of extending life associated with different economic values of life with the payments per life saved at each age. RESULTS: Payments per life saved decreased over time, primarily due to reduced reimbursements. Payments exceeded the age adjusted and the quality-of-life and age-adjusted benefits for all economic values of life at ages 90 and older. CONCLUSIONS: Since the benefits of extending life associated with DRG 475 exceed the payments per life saved at most ages, economic evaluations may be best applied with psychosocial evaluations to allocate resources more ethically. PMID- 10815362 TI - Evaluation of the appropriateness of hip joint replacement techniques. AB - OBJECTIVE: To evaluate the appropriateness of the use of hip joint replacements (HJRs) using explicit criteria developed by an expert panel. METHODS: Observational study. Nine hundred ninety-seven patients from five hospitals with a diagnosis of osteoarthritis, avascular necrosis, hip fracture, or revision who were undergoing HJR were consecutively included in the study during a 1-year period. The appropriateness of the indication was judged by explicit criteria. Complications were recorded at the time of the intervention and 3 months postoperatively. RESULTS: Of the 1,030 interventions, 604 were for osteoarthritis, 31 avascular necrosis, 191 fractures, and 204 revisions. No differences were found among the hospitals for the main clinical and patient variables. Indications for surgery were considered appropriate in 59% of cases, uncertain in 32%, and inappropriate in 8%, mainly in the osteoarthritis group. Differences were found in the rates of appropriateness among some centers. The complication rate did not differ among the groups based on the level of appropriateness of the procedure. CONCLUSIONS: The appropriate use of HJR, as measured by the criteria established by the panel, identified a moderate percentage of inappropriate indications. Those equivocal and inappropriate cases demand further studies to identify patients with an adequate risk-to-benefit ratio from this procedure. PMID- 10815364 TI - Effect of physician specialty on treatment recommendation to patients with coronary artery disease. AB - OBJECTIVE: To assess the influence of physician specialty and the way in which patient data are presented in the treatment recommended for patients with coronary artery disease. METHODS: In a prospective study, 3,628 patients with significant coronary artery disease who had been referred to 1 of 10 heart centers in the Netherlands as possible candidates for either percutaneous transluminal coronary angioplasty (PTCA) or coronary artery bypass graft surgery (CABG) were recruited. Within each center, the recommended treatment is determined by a team consisting of cardiologists only, cardiovascular surgeons only, or cardiologists and cardiovascular surgeons (i.e., composite teams). The main outcome measures are the proportions of patients for whom PTCA, CABG, or noninvasive (medical) therapy was recommended. RESULTS: Composite teams made 71% of recommendations, surgeon-only teams, 12%, and cardiologist-only teams, 17%. Cardiologist-only teams primarily recommended patients to PTCA, surgeon-only teams to CABG, while combined teams made more evenly distributed recommendations (p < .001). Although the patients discussed by the three types of teams were clinically different, the recommendation patterns remained significant after adjusting for these differences (p < .001). For patients with recent myocardial infarction, direct presentation of the case to the team by the referring cardiologist reduced the likelihood that CABG would be recommended. CONCLUSIONS: The treatment recommended to patients with coronary artery disease is affected by the composition of the team providing the recommendation. These findings have important implications for clinical decision making for patients with cardiovascular disease. PMID- 10815363 TI - Factors influencing health insurers' decisions to cover new genetic technologies. AB - OBJECTIVE: To examine the relative importance of factors influencing health insurers' coverage of new genetic technologies. METHODS: A national survey in which the decision makers for private health insurers were asked whether they would cover cystic fibrosis (CF) carrier screening, testing for genetic susceptibility to breast cancer (BRCA test), and medical costs of a clinical trial of gene therapy for CF under a variety of conditions. RESULTS: Respondents' coverage of the two tests and of medical costs of clinical trials was low at the time of the study (4%-15.5% of insurers). Their coverage of CF carrier screening and BRCA testing would be increased significantly if the group tested was restricted to those at high risk, if detection rates were higher and costs lower, and if testing was endorsed by a national professional group or consensus conference. Coverage of the medical costs of a trial of CF gene therapy would be significantly more likely if the trial was restricted to children or adults with severe CF, safety and effectiveness was proven, and therapy could be administered in a regional hospital or an outpatient setting rather than in a research hospital. CONCLUSIONS: Health insurers play a critical role in the diffusion of new genetic technologies. The validity of genetic tests and the safety and effectiveness of new therapies are primary factors influencing health insurers' coverage. Lower costs and approval of professional groups are other factors associated with increased coverage. PMID- 10815365 TI - Extracorporeal shock wave therapy in orthopedics. Assessment of an emerging health technology. AB - OBJECTIVE: Extracorporeal shock wave therapy (ESWT) as an emerging technology in orthopedics has been assessed in Austria with the objective to establish a scientific basis for pending and pressing health policy decisions. Despite encouraging results within some indications and the promising prospect of a noninvasive treatment for some orthopedic diseases, it seemed crucial to assess this new field of application in the light of evidence-based standards, without forgetting the reality of healthcare decisions. This article presents the results of the above mentioned assessment in the context of the policy-making process in Germany, Austria, and Switzerland. METHODS: The study is based on an overview of the already existing reviews on the effectiveness of ESWT as carried out by different actors (health insurances and orthopedists), and on an overview of additional literature covering all relevant aspects (mode of action, recommendations, economic estimates). Orthopedic societies, several institutions involved in health technology assessment, and health insurance agencies were contacted in search of further relevant literature, ongoing studies, and assessments. RESULTS: Until now, clear evidence has not been available on the effectiveness of ESWT in four currently recommended indications (tendinosis calcarea of the shoulder, epicondylopathia humeri radialis, calcaneal spur, and pseudarthrosis), although a substantial body of literature has been produced. Despite encouraging results in some indications, the lack of coherent therapy schemes and the poor quality in the design of clinical studies has been serious enough to hinder the drawing of reasonable conclusions. CONCLUSIONS: Due to the rapid diffusion of ESWT and the pressure on reimbursement agencies to prove the ineffectiveness of ESWT, the policy option of a pragmatic approach is recommended to control the diffusion of an uncertain health technology, alongside active research that would allow a more comprehensive appraisal of this potentially interesting treatment strategy in orthopedics. PMID- 10815366 TI - Implementing clinical guidelines in the treatment of diabetes mellitus in general practice. Evaluation of effort, process, and patient outcome related to implementation of a computer-based decision support system. AB - OBJECTIVES: To evaluate the implementation of clinical guidelines for diabetes mellitus in general practice with a specific computer-based clinical decision support system (CDSS) as part of the intervention. METHODS: Randomized study with health center as unit. General practice in Sor- and Nord-Trondelag counties in Norway, 380,000 inhabitants. Seventeen health centers with 24 doctors and 499 patients with diabetes mellitus were in the intervention group and 12 health centers with 29 doctors and 535 patients were in the control group. Main outcome measures were group differences in fractions of patients without registrations (process evaluation) and mean group differences for the same variables (patient outcome evaluation). RESULTS: Statistically significant group differences were experienced for fractions of patients without registration of cigarette smoking (intervention group, 82.6%; control group 94.5%), body mass index (78.2% vs. 93.0%), and sufficient registrations for calculation of risk score for myocardial infarction (91.1% vs. 98.3%); all during 18 months. Large center variations were shown for all variables. The only statistically significant group difference was 2.3 mm Hg (95% CI, -3.8, -0.8) in diastolic blood pressure in favor of the intervention group. Statistically insignificant differences in favor of the intervention group were HbA1c, -0.1% (95% CI, -0.4, 0.1), systolic blood pressure, -1.2 mm Hg (95% CI, -4.4, 2.0). Statistically insignificant differences in favor of the control group were fractions of smokers, +3.0% (95% CI, -4.0, 10.0), body mass index, +0.3 kg/m2 (95% CI, -0.8, 1.4), risk score in female +0.1 (95% CI, -5.1, 5.2), and risk score in male +2.6 (95% CI, -14.2, 19.5). CONCLUSIONS: Implementation of clinical guidelines for diabetes mellitus in general practice, by means of a CDSS and several procedures for implementation, did not result in a clinically significant change in doctors' behavior or in patient outcome. PMID- 10815367 TI - The use of technologies to minimize exposure to perioperative allogeneic blood transfusion in elective surgery. A survey of Canadian hospitals. AB - BACKGROUND: Despite the growing medical and public interest in reducing exposure to allogeneic blood, little is known about the use of alternatives to allogeneic transfusion. This study was conducted to determine the availability of these technologies in Canadian hospitals and was undertaken under the auspices of the International Study of Peri-Operative Transfusion (ISPOT), a 10-country study of the effectiveness of, attitudes toward, and practices related to the use of alternatives to allogeneic transfusion. METHODS: A cross-sectional national mail survey of Canadian hospitals with greater than 50 medical/surgical beds. Chiefs of anesthesia, surgery, and the divisions of cardiac, orthopedic, vascular, and urology were initially mailed a brief postcard asking which of seven technologies were used in their center. This was then followed up with a one-page questionnaire asking how frequently the technologies were used, their thoughts on the appropriateness of the use of the technologies, barriers to their greater use, and reasons for nonuse of the technologies. RESULTS: Response rates to the postcard survey ranged from 70%-98%, depending on the technology and type of surgery, and ranged from 27%-53% for the follow-up questionnaire. All technologies were used most frequently in cardiac surgery. Aprotinin, tranexamic acid, aminocaproic acid, desmopressin, and cell salvage were reported used in over 70% of cardiac surgery centers. Of these, tranexamic acid and cell salvage were the only ones used routinely in some centers. Acute normovolemic hemodilution and erythropoietin were used in 45% and 20% of cardiac centers, respectively. The drugs were used in less than 15% of orthopedic, vascular, and urologic divisions, with the exception of desmopressin in urologic and vascular surgery and aminocaproic acid in urologic surgery. The techniques of cell salvage and acute normovolemic hemodilution were used in 30%-45% of these divisions, with the exception of cell salvage, which was used in less than 15% of urology units. In more than 60% of cases, the technologies were considered to be used "about right," although an important minority felt that they were underused. CONCLUSIONS: In general, alternatives to perioperative allogeneic transfusion were rarely used except in cardiac surgery. PMID- 10815368 TI - An economic framework for evaluating a multileaf collimator. AB - OBJECTIVES: As health care budgets continue to face close scrutiny, any new acquisition must be evaluated for both costs and outcomes. This study was undertaken to demonstrate the application of an economic framework for the evaluation of a multileaf collimator as an example of a new technology that can have both quantifiable and nonquantifiable benefits for patients, staff, and cancer care institutions. METHODS: Using financial data from the Northeastern Ontario Regional Cancer Centre (NEORCC) and a recognized staffing model, a commercial spreadsheet, developed to economically characterize the principal radiotherapy processes has been used to determine the net incremental annual cost of a multileaf collimator (MLC). RESULTS: The incremental annual cost of purchasing an MLC is estimated at approximately $85,000 (1997 CDN $). Without increasing patient throughput, this increases the average cost of a course of radiotherapy by approximately CDN $200. Savings can be accrued by decreasing mold room activity, increasing the hourly patient capacity on each treatment machine, and decreasing sick time due to strain injuries. CONCLUSIONS: Although the clinical outcome of techniques facilitated by MLCs, such as intensity-modulated radiation therapy, are unknown at this time, an economic context within which to objectively evaluate this technology is presented. The framework presented suggests a method of quantifying outcome-justified expenditures, such as improved patient outcome and greater treatment flexibility, which, when offset against the incremental annual equipment cost, may be used to help justify the acquisition of multileaf technology. PMID- 10815369 TI - Assessing the impact of a consensus conference on long-term therapy for schizophrenia. AB - OBJECTIVE: Our aim was to assess the impact of six recommendations regarding drug prescription on the clinical practices of French psychiatrists. The recommendations were part of the conclusions of a consensus conference entitled "Long-term therapy of schizophrenia" (Paris, January 1994). METHODS: The impact of the conference was assessed on the basis of awareness of the existence of the conference, knowledge of its conclusions, and actual changes in clinical practice. We performed: a) a survey of a representative sample of 396 psychiatrists 2 years after the conference; and b) an analysis of changes in drug prescriptions in a cohort of 2,407 patients with schizophrenia under treatment at the time of the conference. RESULTS: Overall, 78% of interviewed psychiatrists were aware of the existence of the conference and 70% of its conclusions. Declared prescription practices conformed with conference conclusions about 60% (10%-95%) of the time. No difference in practices was noted between psychiatrists who were aware of the recommendations and those who were not. Single neuroleptic prescriptions increased in the cohort study in line with the main conference recommendation. The increase was small, but significant from 51.1% to 56.4%, and mainly concerned patients recently put on treatment. Contrary to recommendations, prescriptions of anticholinergics plus neuroleptics inexplicably rose from 48.2% to 54.3%. CONCLUSION: Small changes in prescription habits occurred in the wake of the consensus conference, but we cannot really ascribe them to a direct impact of the conference. Despite the great pains we took in disseminating the conclusions of the conference as widely as possible, it is clear that a more forceful action plan (e.g., including continuous medical education) is required. PMID- 10815370 TI - Left anterior small thoracotomy versus coronary artery bypass graft for single vessel occlusion. A cost identification analysis. AB - OBJECTIVES: Single-vessel bypass can often be accomplished through less invasive techniques than conventional coronary artery bypass graft (CABG) at substantially lower cost. We undertook a study to empirically determine the cost savings associated with one such technique, left anterior small thoracotomy (LAST). METHODS: Reviewing medical and billing records, we measured the difference in hospitalization costs between two methods of coronary bypass surgery. The study groups consisted of 50 patients who underwent LAST and 28 who underwent single vessel conventional CABG during 1995 and 1996. A subsequent validation sample of 50 patients who underwent LAST was also analyzed. Hospitalization costs were estimated using a relative value unit methodology and were risk-adjusted for both perioperative risk factors and changes in operating room technology. RESULTS: Risk-adjusted hospitalization costs for those undergoing LAST were $9,510 and $12,546 for the CABG control subjects (p < .01), with differences in surgical costs reflecting over 62% of this overall difference. Differences in average length of stay were under a half-day (10.0 for LAST vs. 10.46 for CABG). Only one inpatient fatality was reported; therefore, no inference regarding mortality differences could be made. CONCLUSIONS: LAST is substantially less costly than conventional surgery, and the savings are potentially greater if hospital length of stay is reduced to a clinically recommended time of 2 days. PMID- 10815371 TI - The use of conjoint analysis to elicit willingness-to-pay values. Proceed with caution? AB - There is a growing interest in the potential use of conjoint analysis (CA) as a tool for estimating patient preferences within the economic evaluation of healthcare technologies. It has been suggested that where cost is included as one of the attributes within the exercise, CA can be used to estimate willingness to pay (WTP) for healthcare interventions indirectly within the framework of cost benefit analysis. While recognizing the potential advantages of CA as a technique for the assessment of benefits in health care, this paper recommends several avenues of investigation that need to be carefully explored before an informed judgment can be made as to the validity of eliciting WTP values using the CA methodology. It is argued that much can be learned from the use of qualitative research techniques in examining individuals' understanding and interpretation of CA questions where cost is included as an attribute in the exercise. Additionally, further research is required in defining techniques for establishing the most appropriate levels for the cost attribute in a CA exercise and in determining the sensitivity of WTP estimates to the levels that are chosen for the cost attribute. PMID- 10815372 TI - Using a balancing procedure in multicenter clinical trials. Simulation of patient allocation based on a trial of ventilation tubes for otitis media with effusion in infants. AB - OBJECTIVE: A basic issue in randomized controlled trials (RCTs) is whether we can safely assume comparability between groups at baseline with respect to all potentially important prognostic factors. In other words, did randomization work sufficiently well? In small trials balanced allocation procedures are employed, whereas in large-scale trials simple randomization will do. The question is: When should balancing be considered? METHODS: We performed a simulation study in which we varied the number of categories in the prognostic factors and the number of patients. RESULTS: Simulation showed that, in all instances, a balancing procedure almost always led to perfect or almost perfect balance, while the imbalance with simple randomization was larger. To study the effect of balanced and random allocation on subgroup analyses in our OME trial, we compared the quotient of the width of the confidence intervals (CI). The widest CI in random allocation over the 13 hospitals was on average 13% wider than in balanced allocation. CONCLUSION: Investigators should always consider balanced allocation, especially in categories with a low number of patients and when subgroup analysis over many categories is requested. PMID- 10815373 TI - The European way to health technology assessment. Lessons from an evaluation of EUR-ASSESS. AB - OBJECTIVES: The EUR-ASSESS project was undertaken by a large number of members of European health technology assessment (HTA) agencies and programs with the aim of improving coordination through developments in several areas of HTA. At the conclusion of the project, an independent survey was undertaken to reassess the importance of the original objectives of EUR-ASSESS and determine the impact of the project on the way HTA activities are organized and carried out in Europe. METHODS: A postal questionnaire survey and a series of more in-depth personal interviews. RESULTS: EUR-ASSESS was generally considered successful in achieving its aims, particularly in creating an informal network of people and organizations, improving the understanding of the work of others, improving the ability to focus the objectives of HTA, facilitating the sharing of experience and mutual learning opportunities, exchanging ideas on research agendas, and developing a common language. However, significant differences emerged in the expectations of different actors in European HTA, and in their perceptions of the impact of the project. These reflect the wide diversity of approaches existing in Europe. CONCLUSIONS: Health technology assessment in Europe is changing its nature from a set of national or local initiatives with relatively limited impact to a truly European movement with strong links and coordination between its individual components. Important lessons can be learned from the experience of EUR-ASSESS, particularly in the light of an increasing commitment of the European Commission to play a role in the development of technology assessment. PMID- 10815374 TI - Tumour-associated macrophages (TAMS): disordered function, immune suppression and progressive tumour growth. AB - Evidence currently available suggests that in established, progressively growing solid tumours, tumour associated macrophages (TAMs) are reprogrammed to induce immune suppression of host defenses in situ, through release of specific cytokines, prostanoids and other humoral mediators. This disordered response, results in the inhibition of effective anti-cancer cell-mediated immune mechanisms. Concurrently, TAMs produce tumour growth promoting factors. The summation of this complex interplay of biological factors results in progressive tumour growth and tumour cell dissemination. A better understanding of these complex inter-relationships should form the basis of novel strategies designed to eradicate tumour cells in man and animals. These various biological aspects and processes are discussed in detail and critically evaluated in this review article. PMID- 10815375 TI - Tibial and peroneal artery bypasses using polytetrafluoroethylene (PTFE) with an interposition vein cuff. AB - The combined results of femoro-distal bypasses using prosthetic material with vein cuffs from two separate vascular units is presented. METHOD: Over the last five years, 89 infrainguinal bypasses using polytetrafluoroethylene (PTFE) with a distal interposition vein cuff to a tibial artery have been performed in two centres. All operations were for critical limb ischaemia. The mean age was 72 years (47-90), there were 46 males and 43 females, 27 of whom were diabetic. Twenty-eight patients had a history of angina and/or previous myocardial infarct. The median follow-up was 12 months (0-68 months). RESULTS: During the follow-up period there were 28 deaths, 35 major limb amputations and 44 graft failures. The graft patency rates at 12 and 24 months were 50 and 32%, and limb salvage rates were 53 and 44%, respectively. The patient survival rates were 66 and 61%, respectively. CONCLUSION: The authors believe that the likely improvement in pain free quality of life the patient may enjoy with a successful bypass shifts the balance in some individuals towards bypass surgery using PTFE with a vein cuff to a tibial artery when such an option is possible. PMID- 10815376 TI - Would measurement of C-reactive protein reduce the rate of negative exploration for acute appendicitis? AB - BACKGROUND: Appendicectomy for suspected acute appendicitis is a common procedure. The rate of normal appendices unnecessarily removed remains high (15 30%) despite several techniques and investigations used to improve the diagnostic accuracy. Many studies investigated the role of raised C-reactive protein in improving the diagnosis of acute appendicitis, but with conflicting results. This study emphasies the impact of a normal (rather than raised) serum C-reactive protein in reducing the rate of negative explorations. METHODS: In a double blind study, blood for the measurement of serum C-reactive protein (CRP) was collected pre-operatively from 78 patients just before going to the operating room for appendicectomy. The histopathology of the 78 appendices were grouped into positive (acute appendicitis) and negative (normal appendix). White blood count (WBC), CRP and the histopathology findings were correlated. RESULTS: In patients with histopathologically proven acute appendicitis both the WBC count and serum CRP level were significantly raised (P = 0.025 and P < 0.000,1 respectively). Serum CRP level was normal in 13 out of 15 negative explorations (normal appendix on histopathology). The specificity and sensitivity of serum CRP was 86.6% and 93.6%, respectively. CONCLUSION: A normal pre-operative serum CRP measurement in patients with suspected acute appendicitis is most likely associated with a normal appendix. Deferring surgery in this group of patients would probably reduce the rate of unnecessary appendicectomies. PMID- 10815377 TI - American Society of Anaesthesiologist Physical Status (ASA-PS): a predictor of treatment outcome of transurethral resection of the prostate (TURP). AB - Various factors such as age have been shown to be related to the successful outcome of transurethral resection of the prostate (TURP). Age may not necessarily correlate with the general health and fitness of the patient. A more accurate way to predict outcome of TURP is the pre-operative American Society of Anaesthesiologists Physical Status (ASA-PS) score. This study showed better urological performance, improved QOL and LOS with lower ASA-PS score. It indicates therefore, that ASA-PS is a possible predictor of treatment outcome for TURP. This study indicates a positive correlation between ASA-PS and treatment outcome of TURP. PMID- 10815378 TI - The transabdominal pre-peritoneal (TAPP) inguinal hernia repair: a trip along the learning curve. AB - Laparoscopic inguinal hernia repair is a new but accepted method of treatment for this common condition. It has several distinct advantages over traditional open types of repair but also has the potential for complications not found with open repair. We have performed over 1700 laparoscopic repairs in our unit and modified the technique over the last 5 years whilst auditing our results. We present a description of the technique and describe the common complications we have experienced and the methods we have employed to reduce these complications. PMID- 10815379 TI - Anterior cruciate ligament reconstruction: middle-third bone/patella tendon/bone autograft. PMID- 10815380 TI - Can acute renal failure be prevented? AB - Correction of salt and volume depletion is paramount in the prevention of renal damage. Measures which stimulate intense filtration of glomeruli in acute renal failure, such as the use of atrial natriuretic peptide analogues, theophylline, dopamine, or growth factors should be regarded with caution, since they all increase metabolic workload in the outer medulla and hence aggravate medullary hypoxia. Neither frusemide, dopamine nor dopexamine have been shown to be better than aggressive saline loading in preventing acute renal failure in at risk patients. Until new clinical studies emerge, avoidance of nephrotoxic insults where possible, monitoring of circulating concentrations of potentially nephrotoxic drug levels and volume loading coupled with supportive measures is recommended. When volume depletion persists, usual blood pressure cannot be restored and patients remain oliguric, early referral to the intensive care unit is paramount. The mortality rate in patients with acute renal failure is high; therefore, measures which reduce the incidence and progression of renal dysfunction will be of benefit. PMID- 10815381 TI - The Penman case: a re-evaluation. AB - One of the most remarkable operations carried out in the Royal Infirmary of Edinburgh during the early decades of the 19th century was the surgical excision of an enormous tumour, believed to be an osteosarcoma, of the lower jaw of Robert Penman, that produced severe disfigurement of his face. James Syme, then in his late twenties, was invited to see the patient by Professor Ballingall and Dr John Abercrombie and, in July 1828, he operated, without the benefit of an anaesthetic, to completely remove the tumour. The patient not only survived the operation, but also remained in excellent health for many years afterwards. He emigrated to the United States, but occasionally returned home to Scotland. In 1855, Lord Lister, who noted that his "deformity" had been wonderfully masked by a bushy beard, saw him. This case is re-evaluated in the light of information from the archives in the Royal College of Surgeons of Edinburgh and from the contemporary and more recent literature. PMID- 10815382 TI - Audit of deep wound infection following hip fracture surgery. AB - An audit of 171 consecutive hip fractures treated surgically showed a deep wound infection rate of 3.6%. Several shortcomings in the implementation of the infection policy were identified and recommendations to eliminate them introduced. Review of the subsequent 186 patients gave a deep infection rate of 1.1%. Attention to detail and good practice are essential to maintain low wound infection rates. PMID- 10815383 TI - Information access for the new millennium: publishers and portals. AB - The rise in Internet access has a number of implications for surgeons, both in terms of their personal use of information and in the accessibility of surgical information to their hospitals and patients. This article outlines some of the opportunities offered by the development of new ways to access information, and suggests that the ability to use the new technologies is now becoming an essential clinical skill. PMID- 10815384 TI - Cicatricial pemphigoid: a diagnostic problem for the urologist. AB - We describe a case of cicatricial pemphigoid, which is a chronic bullous disorder that predominantly involves mucous membranes. It can sometimes present to the urologist, with erosions, blisters or ulcers on the male or female genitalia, and can pose a diagnostic problem. The response to medical therapy can be rewarding. However, this can be a chronic debilitating and mutilating disease, and the differential diagnosis should be considered in such cases. PMID- 10815385 TI - Acute small bowel ischaemia complicating emergency colectomy. AB - A case is reported where small bowel ischaemia was precipitated by ligation of the inferior mesenteric artery during emergency colectomy. Aetiology and treatment options are discussed. PMID- 10815386 TI - Subcutaneous splenosis: a clue to diagnosis of thoracic splenosis. AB - We describe a unique case of combined thoracic and subcutaneous splenosis. PMID- 10815387 TI - Acute small bowel obstruction due to ileal endometriosis: a case report and literature review. 1999; 44(1): 59-60. PMID- 10815388 TI - Radiological staging for squamous cell carcinoma of the head and neck: a standardised practice? 1999; 44(5): 303-6. PMID- 10815389 TI - Can the orthopaedic surgeon forget the proctoscope? The proctoscope as tissue protector during intramedullary nailing. 1999; 44(3): 177-8. PMID- 10815390 TI - [Frequency of suicides reduced with 25 percent. Probably by the increased use of antidepressive agents]. PMID- 10815391 TI - [The interplay between embryo and endometrium is important for successful implantation. Increased knowledge can result in new contraceptive methods and better treatment of infertility]. AB - It seems likely that local factors such as cytokines play an important role in synchronizing the development of the embryo and the gestational tract and in determining pregnancy outcome. Evaluation of the precise sequence of cytokines to which the embryo is exposed and responds during the critical periimplantation period may also make it possible to develop culture media which will improve embryo viability and hence increase implantation rates in IVF programs. In addition, an in vitro three-dimensional culture system is being developed which closely resembles endometrium in vivo. This system offers an opportunity to study the local molecular events involved in blastocyst adhesion and to elucidate the role played by local factors, such as cytokines, in producing a receptive luminal epithelium. PMID- 10815392 TI - [Prevalence for clinically proved carpal tunnel syndrome is 4 percent]. AB - This article summarizes the results of a large-scale population-based study conducted to determine the prevalence of carpal tunnel syndrome in the Swedish general population. The study utilized a health questionnaires as well as clinical and electrophysiological examinations. Population prevalence rates of carpal tunnel syndrome, based on clinical diagnosis and electrophysiological criteria, were calculated. Obesity and specific work-related hand activities were shown to be risk factors for carpal tunnel syndrome. PMID- 10815393 TI - [Commission of inquiry scrutinized complications of carotid surgery. Half of the complications could be explicable by errors or dubious indications]. AB - As carotid endarterectomy is a prophylactic procedure, it would seem particularly important to analyze complications with an aim to avoiding them. All carotid endarterectomies in Sweden are registered, and all serious complications (death and permanent neurological deficit) are analyzed in detail, classified and discussed within the profession. During the period 1994-1996 the frequency was 4.3 percent (technical causes in 17 percent, contraindications in 8 percent and dubious indications in 21 percent, but correct indication and surgery in 54 percent). Thus, even when conditions are optimal there is a certain price to pay for the prevention of ischemic stroke. PMID- 10815395 TI - [Postoperative analgesia. Is it time to abandon intramuscular injections?]. PMID- 10815394 TI - [Multidisciplinary cooperation behind a quality registry for carotid surgery. Good coverage during the first two years]. AB - Carotid surgery prevents recurrent stroke in patients with symptomatic tight stenosis of the carotid artery. The Swedish Carotid Surgery Monitoring Registry seeks to promote selection of patients with significant spontaneous risk for recurrent stroke, with an eye toward expediting evaluation and minimizing surgical complications. To this end, professionals at participating hospitals are informed about their own patient selection and surgery risk in comparison with those in the country as a whole and with set targets. We report the results from the first two years, during which it is estimated that more than half of all patients eligible for carotid surgery in Sweden were included in the registry. Although almost all patients had recent onset of relevant neurological symptoms, less than 60% had a documented tight (80-99% occlusion) stenosis. While the final decision to operate a patient was made within 4 weeks of onset of symptoms for only 18% of the patients the first year, this proportion increased to 33% in the following year. The total incidence of surgery related stroke, myocardial infarct and death was 7.7%, while the incidence of severe stroke, myocardial infarct and death was 3.0%. PMID- 10815396 TI - [Treatment of depression and cost efficiency. The cost of a tablet is a poor indicator seen from a socioeconomic perspective]. AB - Direct costs for treating depression, i.e. the cost of out-patient and in-patient care together with drug costs, have increased by 55 per cent during the period 1987-1997 in Sweden. Drugs incurred the greatest increase, whereas the cost of in patient care has decreased. Indirect costs, i.e. sick leave, morbidity and premature mortality due to depression, have also increased during this period. Cost-effectiveness calculations comparing mirtazapine with amitriptyline show that it is less expensive to initiate treatment with mirtazapine, both when direct costs are compared and when indirect costs are included. In a comparison between mirtazapine and fluoxetine, initial treatment with fluoxetine is less expensive with respect to direct cost, but these two alternatives are equivalent when indirect costs are taken into consideration. The price of drug is a poor criterion of resource expenditures and of rational pharmacological therapy in the treatment of depression. PMID- 10815397 TI - [All physicians--not only physicians in charge need leadership training]. AB - Management education programmes for physicians range from university courses to courses offered by private companies. Some hospitals and regions arrange local management education for physicians in specialist training and certified specialists, but mostly for staff in executive positions. Only 245 places are available on the one-week courses for physicians in specialist training during the whole of year 2000 in Sweden; 637 applications were received for the spring courses only. There are 5,000 physicians in specialist training in Sweden. There is a great need for increased investments in the fields of education in administration, economy, leadership and management for physicians. PMID- 10815398 TI - [Production of the most efficient copper intrauterine device has ceased: Will the profit determine women's access to safe contraception?]. PMID- 10815400 TI - [On euthanasia in the Netherlands: don't shoot the pianist--read the books instead]. PMID- 10815399 TI - [Drug therapy of obesity--from bitter experiences to future possibilities]. PMID- 10815401 TI - [Correct information on vitamin B12?]. PMID- 10815402 TI - [B12 deficiency--an underestimated problem]. PMID- 10815403 TI - [The shortage of physicians: private practitioners over the age of 65 are the real manpower reserve]. PMID- 10815404 TI - [Warning against biassed picture news in mass media as evidence of neglect in hospitals]. PMID- 10815405 TI - [Blood transfusion is never completely without side-effects. Filtrated erythrocyte concentrates should be used]. PMID- 10815406 TI - [The first fatal case after gene therapy in the USA. Strong rules guarantee patient safety in Sweden]. PMID- 10815407 TI - [Malpractice in connection with radius fractures must be reduced. Clear guidelines for treatment and follow-up are required]. AB - PSR handles the vast majority of malpractice injuries in Sweden. PSR is a claims handling company which settles claims for malpractice on behalf of the insurance company owned by the Swedish county councils: the County Councils Mutual Insurance Company. A central issue in the law regulating patient injuries in Swedish health care is to define injuries that could have been avoided if a certain therapeutic/diagnostic procedure or a more appropriate method had been utilized. PSR arranged a multiprofessional conference regarding guidelines to decrease the number of malpractice injuries in the treatment of distal radius fractures. Among the most important issues defined were: To improve and standardize diagnostic imaging Patient information Early decision making in surgery and physical/occupational therapy A more well-defined indication for surgery, in which type of trauma, biological age and functional demands are considered in addition to radiographs Less stereotyped thinking in follow-up Controlled randomized trials. PMID- 10815408 TI - [Examination of gastric emptying with gamma camera. Clinically useful, uniform method now established]. AB - Although more than 30 years have passed since the introduction of scintigraphic testing of gastric emptying there has been no well-defined standard. Eight Swedish hospitals have established a nationally standardized method for scintigraphic testing of gastric emptying of solids. 160 healthy subjects participated. The meal consisted of a 99mTc-labeled omelet (1300 kJ) and 150 ml unlabeled soft drink (290 kJ). There were no differences in calculated variables between the centers. Premenopausal women showed slower emptying than postmenopausals and men of any age, making separate reference values for younger women necessary. PMID- 10815409 TI - [Perimyocarditis--a diagnostic dilemma with good prognosis]. AB - Ninety-five patients diagnosed as having perimyocarditis were followed up after a median follow-up time of 66 months. Ninety patients were alive and one patient had died a noncardiac death. Forty-one patients (44%) had leakage of markers of myocardial damage (CKMB or CK) in the acute stage. Seventy-nine patients (83%) were reached for a telephone interview. Sixty patients were in class I, 16 patients were in class II and two patients were in NYHA class III. There was no relation between leakage of myocardial markers and functional class. We conclude that uncomplicated perimyocarditis is a benign disease with excellent prognosis for longterm survival. Leakage of myocardial markers is common, but does not seem to be associated with worse prognosis. PMID- 10815410 TI - [The immune system of the hunter-gatherer meets poverty and excess]. AB - The immune system is closely integrated with the neuroendocrine system, and infection-induced increases in cytokines such as IL-1, IL-6 and TNF have numerous effects on the central nervous system. These include stimulation of the hypothalamus-pituitary-adrenal (HPA) axis, as well as of leptin production. The increase in leptin causes loss of appetite, which may be deleterious for children who are living under conditions of poverty, have frequent infections and are often already undernourished. These cytokines may also be involved in problems of obesity, since they activate the HPA-axis and since TNF is produced by fat cells and can cause insulin resistance. The immune system originally developed for hunter-gatherers may not be well adapted to the pathology of poverty or that of excess. PMID- 10815412 TI - [Sex cases of sudden death after snorting heroin analyzed. Reduced tolerance in periodic abuse a danger both in snorting and injecting]. AB - Heroin injection entails a risk of infection and can result in sudden collapse and death. During the past few years alternative routes of heroin use have been introduced. During 1997-1998 we observed six deaths which occurred suddenly following heroin snorting. These victims were all temporary drug users in good health and physical condition. They all died with low morphine concentrations; however, three had relatively high blood alcohol concentrations and two were under the influence of medicinal drugs. Temporary use of heroin is characterized by low drug tolerance; snorting of heroin appears to entail the same risk of sudden death as injection. PMID- 10815411 TI - [Buprenorphine as a new alternative for detoxification of heroin addicts. It causes only mild withdrawal problems, abating quickly]. AB - Buprenorphine might be an alternative drug to be used in opiate detoxification. Its main advantage is that it carries a low risk for respiratory depression, it gives less euphoria and limited withdrawal effects. In a pilot detoxification project ten heroin addicts were given buprenorphine; seven completed the course. Before detoxification seven patients showed five or more signs on the Himmelsbach scale [6]. After the second day four patients showed no signs, four patients displayed one sign, two patients two signs. Buprenorphine may be a valuable alternative to clonidine, dextropropoxiphene and methadone in the detoxification of opiate addicts. PMID- 10815413 TI - [A promising model for geriatric psychiatry. Integrated outpatient and inpatient care, based on the patient's health]. PMID- 10815414 TI - [Return of malaria, outbreaks of dengue fever. Possible health effects of climate changes in Europe, according to the WHO]. PMID- 10815415 TI - [Sigmund Freud discovered the therapeutic effects of cocaine, but all the credit went to Carl Koller]. PMID- 10815416 TI - [Moment in medicine, part 5. Is kindness a must for a physician?]. PMID- 10815417 TI - [This is for primary health to take care of...]. PMID- 10815418 TI - [Physicians employed by the insurance authority take their responsibility. We don't oppose scrutiny by a supervising authority]. PMID- 10815419 TI - [Disabled pupils are short of time! The compulsory school period should be prolonged with 3 years]. PMID- 10815420 TI - [Percutaneous endoscopic gastrostomy (PEG) in children--a reply]. PMID- 10815421 TI - [Misleading concentration of ethanol in urine from rape victims with diabetes mellitus]. PMID- 10815422 TI - [Examination of 5-year old children as an "alphabetization campaign"]. PMID- 10815423 TI - A century of service in medicine. A history of the Michigan State Medical Society. PMID- 10815424 TI - William Beaumont, M.D., and his studies of the physiology of digestion at the Michigan Gibraltar of the Great Lakes. PMID- 10815427 TI - 1932-34 was busy time for society with state and federal legislation PMID- 10815425 TI - MSMS faces World War I, other pressures and problems during 1915-1929 period PMID- 10815426 TI - 1929-33 recall problems caused by depression; joint committee builds up public opinion PMID- 10815428 TI - Ambitious public education program helped tell medicine's story PMID- 10815429 TI - Economic problems command considerable attention PMID- 10815430 TI - Governor Murphy signs Basic Science Bill PMID- 10815431 TI - Doctor placement service inaugurated PMID- 10815433 TI - MSMS diamond jubilee PMID- 10815432 TI - House of Delegates in special session approves group medical, hospital service PMID- 10815434 TI - Efforts centered on war and defense PMID- 10815435 TI - Michigan Health Council is organized PMID- 10815436 TI - Michigan Foundation for Medical and Health Education launched PMID- 10815438 TI - Honor U-M medical school centennial PMID- 10815437 TI - First rural health conference sponsored PMID- 10815439 TI - Funds contributed for Beaumont Memorial PMID- 10815441 TI - Busy years for scientific, PR activities PMID- 10815440 TI - Beaumont Memorial dedicated PMID- 10815442 TI - MCI telecasts, seal of assurance in the spotlight PMID- 10815443 TI - Construction of headquarters completed PMID- 10815446 TI - Centennial session plans make year a busy one PMID- 10815444 TI - Presidents program, McNerney report in headlines PMID- 10815445 TI - Mr. Brenneman chosen to succeed Mr. Burns PMID- 10815447 TI - The privilege of making history. PMID- 10815448 TI - [Food allergies]. AB - Clinical aspects of food allergies in infants and children are discussed together with diagnostic procedures in order to provide for practitioners a better understanding thus allowing a good management of pediatric patients with food related disorders. PMID- 10815449 TI - [Some current problems in pediatric otorhinolaryngology]. PMID- 10815450 TI - [Nutritional information for children to modify the food habits of the whole family]. AB - The prevention of weight gain and cardiovascular diseases is probably easier, less expensive and more effective than treating these diseases after they have fully developed. The hypothesis of a prospective intervention study in northern France, the "Fleurbaix Laventie Ville Sante Study" is that nutritional education of children aged 6-12 years at school may not only improve their nutritional knowledge but also influence the dietary habits of the family. We have conducted an information program in school for these children and made an evaluation of the educational program used in this study and the modification of the dietary habits. The first results indicate a better nutritional knowledge in children of Fleurbaix and Laventie versus control children. The families have better dietary habits consuming less fat, more carbohydrates. In women the BMI is less important in Fleurbaix and Laventie compared with the control subjects. This study is on going with a 10 year-follow up taking into account the spontaneous evolution in a control town. PMID- 10815451 TI - [Divorce and new families: strategies for preserving the bonds]. PMID- 10815452 TI - [Strategies for preserving the bonds: Meeting Points]. PMID- 10815453 TI - [The family and the pediatrician, synergy and complementarity in a systemic approach in pediatrics]. PMID- 10815454 TI - [Congenital nephropathies]. AB - The early diagnosis of congenital nephropathies, sometimes before birth, is mandatory for the optimal management of affected children. While they do not always modify the final outcome of the disease, preventive measures can attenuate the consequences, and sometimes significantly delay the progression of the nephropathy. This paper presents a simple classification of congenital nephropathies, and briefly describes their mode of presentation. PMID- 10815455 TI - [Vesico-ureteral reflux in children: current knowledge and management]. AB - Reflux nephropathy is an important cause of chronic renal failure and arterial hypertension in children. While its prognosis is favourably influenced by the early recognition and management of vesicoureteric reflux, there is a real risk of missing the diagnosis of this insidious condition. This article describes the main features of vesicoureteric reflux and associated nephropathy and presents practical recommendations, through a nonexhaustive literature review. As several aspects of this disease remain to be established, any recommendation concerning its management should be constantly reevaluated according to new data. PMID- 10815456 TI - [Proteinuria in children: practical approach]. AB - Significant proteinuria is not an unfinding in children. Its causes are variable. When detected by dipstick examination of urine, the proteinuria must be assessed quantitatively by measuring the urinary protein/creatinine ratio in a spot sample. Orthostatic proteinuria is the most common cause of intermittent proteinuria. Persistent glomerular or tubular proteinuria are the consequences of various glomerulopathies or tubulopathies, the prognosis of which is variable. Whether glomerular or tubular, persistent proteinuria must be fully investigated, including by renal biopsy. PMID- 10815457 TI - [Investigation of the urinary tract in children in nuclear medicine]. AB - The early detection of urologic abnormalities by antenatal sonography has resulted in the investigation of many infants and neonates for suspicion of either obstructive uropathy or reflux nephropathy. Nuclear medicine techniques allow to assess renal parenchyma integrity, to detect pyelonephritic scars and to measure absolute and relative renal function; these methods are easy to perform and reproducible, without any sedation, repeated venous punctures or bladder catheterization. Furthermore, the use of dynamic tubular tracers and frusemide test is a very usefull method which can differentiate between upper and/or lower urinary tract obstruction and determine the degree of obstruction (severe or incomplete) in order to plan for surgery or conservative treatment. The detection of vesicoureteric reflux may be difficult as it is an intermittent phenomenon: the use of the indirect radionuclide cystography (IRC), that is to say after completion of a dynamic renography, allows to detect reflux with a high sensitivity because images can be recorded continuously until the child voids, without any bladder catheterization and at low radiation dosis. In case of discordant results between micturating cysto-urethrography and IRC or of concomittant obstructive uropathy, the direct radionuclide cystography (DRC) is indicated for appropriate treatment. Nuclear medicine techniques do not give morphological information about the urinary tract and should be considered as complementary to radiological investigations in first evaluation of children with recurrent urinary tract infections or hydronephrosis. PMID- 10815458 TI - [Carbon monoxide poisoning in children: never trivialize]. AB - The risks linked to tissular hypoxemia after carbon monoxide (CO) poisoning are well known. Unawareness of CO exposure and of its complex pathophysiology may delay appropriate treatment and lead to long term neuropsychological sequelae. We report two cases of children treated in our institution and review the main issues regarding the optimal management. A high index of suspicion for carbon monoxide poisoning when dealing with an unclear neurological clinical presentation is mandatory. Classical therapy with normobaric 100% oxygen has to be instaured immediately. Hyperbaric oxygen therapy must be considered when anamnestic symptoms or clinical signs suggest neurological involvement even when carboxyhemoglobin values are low or already normalized. PMID- 10815459 TI - [Atopic dermatitis: the first allergic step in children]. AB - Atopic dermatitis is an early manifestation of atopy and is frequently present already during the first year of life. Atopic dermatitis is often associated with increased levels of serum IgE and with sensitization to food allergens. Some foods may be responsible for exacerbations of skin inflammation, but their pathogenic role need to be clinically assessed before an avoidance diet is prescribed. Staphylococcus aureus, which is known to colonize the skin of atopic dermatitis patients, may also exacerbate skin lesions and need to be treated with topical antibiotics. Children with atopic dermatitis are prone to allergies and are at risk to develop later respiratory allergic manifestations. In particular, if a sensitization to egg is present early in life, the risk for developing later an asthma due to house dust mites is increased. The care of children with atopic dermatitis should not be limited to the treatment of skin lesions, but should also involve preventive measures for respiratory allergies. PMID- 10815460 TI - [Sudden infant death: a drama in spectacular regression]. PMID- 10815461 TI - [Auditory screening in school. Comparison of predictive values with vision screening]. PMID- 10815462 TI - [What is your diagnosis? Bilateral abscess forming pneumonia]. PMID- 10815463 TI - [Rosacea, acne and other diseases of the seborrheic spectrum]. AB - Diseases of seborrhoic origin include rosacea, acne, gram-negative folliculitis, demodex-folliculorum, perioral dermatitis as well as seborrhoic dermatitis. An important prerequisite for adequate therapy is the knowledge of these different diseases. Unfortunately, topic steroids are often applied, resulting in well known corticosteroid side-effects like skin atrophy and teleangiectasias. The different clinical diseases of seborrhoic origin are reviewed here and a treatment guide is presented. PMID- 10815464 TI - [Peracute suffocation emergency: what should be done?]. AB - Peracute obstructive dyspnea with imminent asphyctic death is a dramatic emergency. It is characterized by easily audible inspiratory stridor caused by severe stenosis. Several causes in the pharynx and larynx may be responsible. Pharmacotherapy is hardly effective, an intubation extremely difficult in extremis. In contrast emergency coniotomy offers a real chance to reopen the airways even to the surgically inexperienced. The technique for this intervention is described. PMID- 10815465 TI - [Drug therapy of obesity. Strategies for general practice]. AB - The pathogenesis of obesity is complex and miscellaneous endogenous as well exogenous factors modulate body weight. The therapeutic strategies are equally complex and may include the option of antiobesity drugs. Despite the availability of drugs they are only seen as adjuvant therapeutic options and the corner stone of obesity therapy remain permanent life style changes. Presently only orlistat and sibutramin are approved for the pharmacological therapy of obesity in Switzerland. The indications and contraindications of these new antiobesity drugs are discussed. In addition a new concept of a stepped care approach in weight management is presented. PMID- 10815466 TI - [Leukotriene antagonists--do they have a value?]. AB - Guidelines on the treatment of asthma emphasize the importance of long-term control of the disease. The leukotriene receptor antagonists are a new class of maintenance therapy for the treatment of chronic asthma. In Switzerland two of these receptor antagonists are available, zafirlukast (Accolate) and montelukast (Singulair). Compared with placebo leukotriene antagonists are active and improve mild-to moderate asthma in terms of improving lung function and symptoms and decreasing exacerbations. However compared to low dose inhaled steroids LTRA are less effective and more expensive. Inhaled steroids therefore remain the preferred and most cost-effective therapy for treating chronic asthma. Among patients requiring moderate to high doses of inhaled corticosteroids to maintain asthma control LTRA allow for reduction of the doses. The best response to LTRA show patients who have aspirin-sensitive asthma. PMID- 10815467 TI - [De Quervain subacute thyroiditis. De Quervain subacute thyroiditis DD: Hashimoto thyroiditis]. PMID- 10815468 TI - [The future is not as it used to be]. PMID- 10815469 TI - [The future is light--there are only three problems]. PMID- 10815470 TI - [Family practice at the turn of century--on the way down or new possibilities?]. PMID- 10815471 TI - [Resuscitation of newborn infants]. PMID- 10815472 TI - [Infrared tympanic thermometry compared to mercury thermometers]. AB - Because of conflicting reports we examined the accuracy of infrared tympanic thermometry compared with mercury rectal thermometer and a digital rectal thermometer in a medical ward. We studied prospectively the accuracy of parallel measurements with infrared tympanic thermometer and the correlation between this method and the rectal mercury thermometer. Measurements with digital rectal thermometry are also compared with mercury thermometry. 191 adult inpatients were included. The median difference between infrared tympanic and rectal mercury thermometry in the whole material was -0.5 degree C, but increased to -1.4 degrees C in a selected group with rectal temperature 38 degrees C or more. The median difference between parallel measurements with infrared tympanic thermometer was 0.4 degree C. Digital rectal thermometry, however, was in strict accordance with the rectal mercury method. We found an unacceptable difference in body temperature between measurements with infrared tympanic thermometer and rectal mercury thermometer. In a clinical setting the infrared ear thermometer has a very low sensitivity for detecting fever. Digital rectal thermometry seems to be a good alternative to the rectal mercury thermometer. PMID- 10815473 TI - [Traditional surgery of inguinal hernia]. AB - Inguinal hernia repair is associated with a variety of complications of which the recurrence rate is one of the most important. The aim of this study was to examine all patients one year after surgery for inguinal hernia in an outpatient clinic. 12 months after the operation patients were interviewed and physically examined. 172 hernias in 166 patients were repaired in 1996 in our outpatient unit. 17 surgeons used nine different techniques. Follow-up was achieved in 93.9% of patients alive. Ten recurrences were found (6.3%). Four of the patients were not aware of their recurrence. 11% of the patients still had discomfort or pain after twelve months, whereas 5.2% had not resumed full activity. Other complications occurred in more than 20% of the operations. A "free to do policy" in inguinal hernia repair results in high recurrence and complication rates even in the hands of experienced surgeons in our clinic. The present study shows that an interview as well as a physical examination are necessary in evaluating recurrences, postoperative complications, postoperative pain and time to full mobilisation. Without physical examination, the recurrence rate would have been underreported. PMID- 10815474 TI - [Resuscitation of newborn infants with room air or oxygen]. AB - In this article we present results from the Resair 2 study in which we tested whether room air is more efficient than 100% oxygen in newborn resuscitation. Eleven participating centres in Egypt, Estonia, India, Norway, Philippines, and Spain recruited 609 infants who needed resuscitation at birth; of these, 288 were resuscitated with room air and 321 with 100% oxygen. There were no differences between the two groups with regard to outcome. One minute Apgar scores were significantly lower in the oxygen group than in the room air group. Median time to first breath was significantly delayed with 24 seconds in the oxygen group compared with the room air group. It seems that 100% oxygen depresses ventilation in newborn infants. Room air is as safe and efficient as 100% oxygen at least in most cases of newborn resuscitation. Further studies confirming these results are needed before new resuscitation routines are implemented. PMID- 10815475 TI - [The senile immune system]. PMID- 10815476 TI - [Guidelines for resuscitation of newborn infants]. AB - This article presents the new guidelines for resuscitation of the newborn from the European Resuscitation Council. It is estimated that, potentially, 800,000 newborns can be saved each year by simple airways manoeuvres. Personnel trained in basic resuscitation should be present at all deliveries and personnel trained in advanced resuscitation at deliveries with known risk factors. Attention to ventilation is of primary importance. Ventilation should be assisted if stimulation does not achieve a prompt onset of spontaneous respiration and/or the heart rate is less than 100 per minute. Chest compressions--1/3 of the anteroposterior diameter of the chest--should be provided if the heart rate is absent or remains < below 60 per minute despite adequate assisted ventilation for 30 seconds. Chest compressions should be coordinated with ventilation at a ratio of 3:1 and a rate of 120 "events" per minute to achieve approximately 90 compressions and 30 rescue breaths per minute. Adrenalin should be given if the heart rate remains below 60 per minute despite 30 seconds of effective ventilation and chest compression. PMID- 10815477 TI - [Guidelines for cardiopulmonary resuscitation of children]. AB - In this article the European Resuscitation Council's guidelines for basic and advanced resuscitation of children are presented. There are some changes from the previous guidelines. Children are divided in three age categories (in addition to the newly born): children up to one year old, one to eight years old, and more than eight years old. In Norway, but not in the rest of Europe, evaluation of the circulation by pulse check has been eliminated in basic, but not in advanced resuscitation. This is due to reports that pulse checks by lay rescuers require much time with poor specificity and sensitivity. In evaluating the patient's own ventilation, the differentiation between agonal gasps and regular breaths is stressed. ECG monitoring provides the link between paediatric basic life support and advanced life support. The algorithm for the latter closely resembles that proposed for adults. While there were previously three separate algorithms for ventricular fibrillation/ventricular tachycardia, asystole and electromechanical dissociation, there is now only one algorithm. Ventilation and chest compressions should be performed for one-minute periods with ventricular fibrillation/ventricular tachycardia, for three-minute periods with other rhythms. PMID- 10815478 TI - [DNA ploidy in epithelial ovarian cancer--an independent prognostic factor]. AB - Ovarian cancer is the most lethal of the gynecological malignancies. One of the aims of the ongoing research in this field is the search for prognostic and/or predictive factors which can contribute to a more individualized patient treatment. All studies performed at The Norwegian Radium Hospital with regard to the prognostic significance of DNA ploidy in borderline, early and advanced ovarian cancer, were reviewed. The conclusions emanating from these studies were compared to the international literature. DNA ploidy analysis is of definite independent prognostic significance in borderline and early (FIGO stage I) ovarian cancer, and is of help in the selection of patients expected to benefit from adjuvant chemotherapy, or in whom a more conservative surgical procedure can be acceptable. DNA ploidy status is also of prognostic significance in advanced ovarian cancer; however, for the time being this information has no direct consequences for patient treatment. We conclude that DNA ploidy analysis should be incorporated in the routine histopathological evaluation of borderline and early (stage I) ovarian cancer. PMID- 10815479 TI - [Quality of life, loneliness and social contacts among persons with long-term mental illness]. AB - Chronic long-term patients who were residents in psychiatric nursing homes at a given point in time were traced six years later. Over this period, there had been deep cuts in the number of psychiatric beds in the county in question; we wanted to assess how the patients now perceived their quality of life. All 107 patients were traced; of the 75 who were still alive, 74 took part in the study and were visited at their place of residence. 42 patients, mean age 56.9 years, were able to respond to personal questions regarding social contact, loneliness and quality of life. Health care providers were the most important persons in the patients' networks. Most patients reported a satisfactory quality of life; those who lived outside institutions (N = 21) tended to be more satisfied than those in residential care, they were more socially active and had better contact with their families. The variables loneliness, satisfaction with neighbourhood and leisure activities explained 63% of the variance in quality of life. PMID- 10815480 TI - [Emergency psychiatric departments are not refusing admissions]. AB - The staff at the psychiatric emergency out-patient clinic in Oslo evaluated the services of the acute psychiatric wards when admitting patients to the hospital. Of 93 admittances during three months in 1998, the referring doctor was satisfied with the hospital's response in 84% of the cases. The mean time from contact was taken with the hospital till the admittance was accepted was 19 minutes. The admittance of patients in acute psychiatric hospitals in Oslo is straightforward, and reasonably well organized. PMID- 10815481 TI - [Will the revolutionary knowledge of apoptosis play an important role in medicine?]. AB - Apoptosis is a phenomenon of fundamental importance in embryonal development and the homeostatic regulation of mature tissue. We review the present knowledge on apoptosis and the evidence that apoptosis may play a role in the pathogenesis of human disease. As examples we discuss its role in cancer, ischemic diseases, heart failure and neurodegenerative diseases. PMID- 10815482 TI - [The mechanism of apoptosis--therapeutic possibilities in cancer]. PMID- 10815483 TI - [The mechanism of apoptosis--significance for heart failure?]. PMID- 10815484 TI - [Glutamate, apoptosis and necrosis of the nervous system]. PMID- 10815485 TI - [Image guided and robotic treatment--the advance of cybernetics in clinical medicine]. AB - The introduction of advanced technology in hospitals has changed the treatment practice towards more image guided and minimal invasive procedures. Modern computer and communication technology opens up for robot aided and pre-programmed intervention. Several robotic systems are in clinical use today both in microsurgery and in major cardiac and orthopedic operations. As this trend develops, professions which are new in this context such as physicists, mathematicians and cybernetic engineers will be increasingly important in the treatment of patients. PMID- 10815486 TI - [Who directs the medical development?]. PMID- 10815487 TI - [Medical technology--a friend or an enemy?]. PMID- 10815488 TI - [Control technology results in less control]. PMID- 10815489 TI - [From ready-made to tailor-made--future possibilities for individualized drug therapy]. AB - Knowledge of the genetic code offers great promise for individualised drug therapy. Recent advances in molecular biology and biotechnology have led to new possibilities in the prediction of therapeutic effects and risks of adverse reactions based upon the patient's individual genotype. In addition, these techniques are the basis of gene therapy and are a prerequisite for the development of many other new drugs. This article presents the current status in these fields and discusses to which extent the present promises can be expected to be realised in the future. PMID- 10815490 TI - [Tailor-made drugs in the new millennium]. PMID- 10815491 TI - [Who will receive future tailor-made drugs?]. PMID- 10815492 TI - [Global trends in food consumption and nutrition]. AB - Obesity and lifestyle diseases increase all over the world, especially in developing countries. One reason is the change in diet. This nutrition transition is characterised by improvement in dietary variation, but also by increase in the content of fat and sugar. The transition seems to start at a lower level of income, compared to what occurred in the Western countries after the Second World War. The reason is that many foods are relatively cheaper, especially fat and sugar. The world market is presently flooded with cheap vegetable fat. Urbanisation leads to over-consumption by increasing market access to fatty and sugary foods, including fast foods. Globalization increases the consumption of sweet soda pops, biscuits and snacks produced by multinational companies. Western supermarkets and fast food franchises also promote these dietary changes (McDonaldization). It has been proposed that the population in developing countries is more vulnerable towards these dietary changes in regard to obesity and chronic diseases, due to undernutrition in early life (the Barker hypothesis). We may therefore expect an unprecedented increase in the prevalence of chronic diseases, especially diabetes type 2 in the developing countries. One may question if this increase will be a transient phenomenon, or if we can expect the same pattern as we have seen in the West, namely that the poor become the fat test segment of the population, with the highest prevalence of chronic diseases. PMID- 10815493 TI - [Is it safe to eat food?]. PMID- 10815494 TI - [A society not friendly to physical activity?]. PMID- 10815495 TI - [More women in the medical profession--a benefit?]. AB - More than a hundred years have passed since the first woman graduated in medicine in Norway. Today, more than 30% of doctors and 20% of specialists are women. In some specialties there are still few women: in general surgery 5%, in otorhinolaryngology 9%; only in some 10% of the specialties are there a little above 30% female doctors. There are also few women in positions of leadership. A better balance between men and women would be a benefit to the medical profession, to the patients, to the working environment and in the shaping of the politics of healthcare. There is a change going on, but a slow one. At present, 50% of medical students are women. It is to be hoped that they will be able to make their career choices on the basis of their skills and preferences rather than their gender. Women doctors make a positive contribution to a male-dominated profession; indeed, they may contribute to a development and expansion of the doctor's role. This calls for changes in attitudes, good support and role models- and satisfactory working conditions. Project groups within the Norwegian Medical Association as well as its Equal Opportunity Committee will be working for this in the new millennium. PMID- 10815497 TI - [Equality takes time--a lot is left to do]. PMID- 10815498 TI - [Medical progress--myths and realities]. PMID- 10815499 TI - [Medical technology in the new millennium]. PMID- 10815500 TI - [Medical progress requires respectful physicians]. PMID- 10815501 TI - [Scenario analysis--a method for long-term planning]. AB - Scenarios are known from the film industry, as detailed descriptions of films. This has given name to scenario analysis, a method for long term planning using descriptions of composite future pictures. This article is an introduction to the scenario method. Scenarios describe plausible, not necessarily probable, developments. They focus on problems and questions that decision makers must be aware of and prepare to deal with, and the consequences of alternative decisions. Scenarios are used in corporate and governmental planning, and they can be useful and complementary to traditional planning and extrapolation of past experience. The method is particularly useful in a rapidly changing world with shifting external conditions. PMID- 10815502 TI - [13 years old scenarios for this year]. AB - This article examines two scenarios, published in 1987, for the future development of Norwegian health care towards the year 2000. The scenarios emphasize different external forces and priorities. The 1987 scenarios are then assessed in the light of actual developments during the period. Scenario I describes a picture characterised by reduced economic growth 1987-2000 compared to previous decades, resulting in a year 2000 health care system with limited resources and hard priorities, focus on cost containment, increased efficiency, and search for alternative solutions. In Scenario II, the year 2000 health care system had more resources; however, the demand for services was as high or higher than in the 1980s. Instead of improvements in efficiency, alternative modes of financing health care and social security had evolved. Increasing knowledge among consumers, dissatisfaction and new technology had driven up costs, and health care professions tried to take advantage of the situation. This scenario is characterized by people wanting more, and getting more. PMID- 10815503 TI - [Too few physicians or too many patients?]. PMID- 10815504 TI - [Health services needs--naturally or socially based?]. PMID- 10815505 TI - [BiliBed and treatment of neonatal jaundice]. PMID- 10815506 TI - [Nitric oxide]. PMID- 10815507 TI - [Intracranial arachnoidal cysts]. PMID- 10815508 TI - [Allergic reactions during anesthesia--a national network for registration, diagnosis and follow-up]. PMID- 10815509 TI - [Are there any healthy Norwegians?]. PMID- 10815510 TI - [Emergency medical service]. PMID- 10815511 TI - [DNA tests, Somalis and Norwegian adoptive parents]. PMID- 10815512 TI - [Research and specialists]. PMID- 10815513 TI - [Screening for lung cancer]. PMID- 10815514 TI - [Screening for lung cancer. Promising results with low-dose CT]. AB - Low-radiation-dose computed tomography (low-dose CT) is a new, promising technology presently used in screening for lung cancer in some medical centres in the USA and Japan. The three population-based studies of the efficacy of screening with low-dose CT published to date all show that more than 85% of the lung cancers detected by low-dose CT are in stage I, offering improved possibility for curative treatment. The number of false positive scans, was up to 20%. The radiation dose per low-dose CT scan was approximately the dose of a mammography (1 mSv). In two studies, low-dose CT failed to detect some centrally located tumours. None of the studies are capable of evaluating changes in lung cancer mortality in a screened population compared to those of a control population. There is, therefore, a need for further assessments of positive as well as negative effects of population-based screening with low-dose CT. PMID- 10815515 TI - [Porphyria cutanea tarda]. AB - Porphyria cutanea tarda (PCT), the most common porphyria disease, is characterized by blistering and skin fragility of sun-exposed skin. The symptoms are caused by lowered activity of uroporphyrinogen decarboxylase (URO-D) resulting in accumulation of water-soluble porphyrins in the skin. Most PCT cases are sporadic but can be familiar due to mutations in the URO-D gene located on chromosome number 1. The disease may be exacerbated by environmental factors. Iron accumulation is a characteristic finding and there is an association to hereditary haemochromatosis. Therapeutic venesection reduces the iron load and the uroporphyrins are mobilized by treatment with hydroxychloroquine. An increased risk of liver cirrhosis and hepatocellular carcinoma may presumably be reduced by early diagnosis and treatment. PMID- 10815516 TI - [Prehospital cardiac arrest in Denmark. Are emergency services efficient?]. AB - The chain-of-survival concept is an established principle in the treatment of out of-hospital cardiac arrest. Survival is improved when out-of-hospital cardiac arrest is followed by a sequence of events: early access to help, early cardiopulmonary resuscitation, early defibrillation and early advanced care. The Danish emergency medical system is only to some extent in accordance with the standards of the chain-of-survival concept. The current way of evaluating the efficiency of the system is based mainly upon the driving time for the ambulance, but the time that counts for the patient is the time between the emergency call and the start of adequate treatment. Options for defibrillation and--in some cities--advanced life support are good, but hard to exploit as ambulances frequently arrive too late for treatment to be effective. PMID- 10815517 TI - [Use of antibiotics among Danish children in 1998. Consumption on a national basis]. AB - The aim was to evaluate the use of antibiotics (AB) among Danish children in 1998. Patient specific data was collected on antibacterials for systemic use (ATC group J01) prescribed for children aged 0-15 years in 1998. Data included a total of 585,491 prescriptions for 331,979 individuals. AB were prescribed for 33% of all Danish children on one or several occasions in 1998. Younger children were more likely to be treated with AB than older children. The one-year-prevalence of the use of AB was 63.2% for the 1-year-olds and 18.3% for 12-year-old children in 1998. The type of AB used varied with age, with younger children more prone to receive broad-spectrum AB. Eighty-three percent of all AB-prescriptions were prescribed by general practitioners. It is concluded, that AB, especially the broad-spectrum AB, are more often prescribed for younger than for older children and adults in Denmark in 1998. PMID- 10815518 TI - [Are out of hours contacts with physicians necessary? Assessment by physicians on call and by patients]. AB - Patients and GPs were asked about necessity and possible prevention of contacts to the out of hour service. 4187 (83.9%) of the GP questionnaires and 1493 (52.3%) of the patient questionnaires were returned. According to the GPs 25% of the contacts were not necessary and 5% were not necessary according to the patients. Patients and GPs agreed that contacts concerning old people were most necessary. GPs assessed contacts concerning children relatively more necessary. Patients assessed contacts concerning middle-aged relatively more necessary. Frequent users were assessed less necessary according to GPs but more necessary according to the users themselves. Many contacts could have been prevented according to both GPs and patients--especially within the first five hours of the service. Concordance between a patient's and a GPs answer was bad (kappa = 0.0 0.2). Concordance between the GPs answering the phone and the GPs examining the patient was only slightly better (kappa = 0.1-0.3). PMID- 10815519 TI - [Big differences in the frequency of polypharmacy between physicians]. AB - Polypharmacy, the simultaneous use of multiple drugs, is associated with adverse drug reactions, medication errors, and increased risk of hospitalisation. When the number of drugs is five or more (major polypharmacy), a significant risk may be present. We analysed the prevalence of major polypharmacy among listed patients, and identified possible predictors of major polypharmacy related to the practice. Prescription data were retrieved from the Odense Pharmaco epidemiological Database and the age and sex standardised prevalence rate of major polypharmacy was calculated for each practice (n = 173). Possible predictors of major polypharmacy related to the GPs were analysed by backward stepwise linear multiple regression. A six-fold variation between practices in the prevalence of major polypharmacy was found, i.e. from 16 to 96 per 1000 listed patients (median: 42). Predictors related to the practice structure, workload, clinical work profile, and prescribing profile could explain 56% of the variation. PMID- 10815520 TI - [Differences when it comes to induced abortion in different Danish counties]. AB - The paper describes actual regional differences in the rate of legally induced abortions as well as different trends observed since abortion on request was introduced in 1973. The analysis is primarily based on national population-based and published data from the Registry of Legally Induced Abortions in the National Board of Health, supplemented with data from a linking with the Fertility Database. The rate of abortion has shown regional differences, being at the highest level in counties with large cities. The observed differences are related to higher age at first birth and fewer children per woman in counties with a high frequency of abortion. A large part of the differences are observed among women below age 30-35. Over time developments have shown a parallelism between the counties, even though some differences have also been observed as regards both induced abortion and fertility. Various background factors do not seem to have the same impact on the choice of induced abortion in all counties. PMID- 10815521 TI - [Proper advice on nutrition. Recommendations concerning the conduct of the nutritional news]. AB - The Committee on Proper Conduct in Public Advice on Nutrition, set up by the Danish Nutrition Council, has produced recommendations to persons involved in communicating results of nutrition research. The guidelines are targeted on scientists, industry, journalists and publishers as well as consumers to facilitate proper conduct in communication of own or other scientists' research results, which might influence consumers' choice and intake of food. The results are presented as recommendations and checklists providing the necessary requirements to bring people in a position to make the best possible evaluation of the new scientific results and to put the results into their proper perspective. The aim of this report is to improve the process so that the communicator focuses on the background information necessary for the reader/listener/viewer in the current situation to achieve a balanced view and benefit from the new scientific result. PMID- 10815522 TI - [Asymmetric periflexural exanthema of childhood]. AB - Asymmetric peri-flexural exanthem of childhood is a "new" disease, which has never been reported by Danish authors. It is characterized by an asymmetric maculopapular exanthem in early childhood. The exanthem is initially unilateral and localised close to the axilla, but it spreads centrifugally during the first week, becomes more widespread and resolves spontaneously within four to six weeks. We describe two cases of this entity. PMID- 10815523 TI - [Intraspinal melanocytoma in a 16-year old girl]. AB - Meningeal melanocytomas are rare benign neoplasms of meningeal melanocytes, and are most frequently located in the posterior fossa and along the cervical spinal cord. We report a case of a 16 year-old girl with a melanocytoma located in the upper cervical part of the spinal canal. The patient presented severe neurological signs, but had a very good outcome following surgery. PMID- 10815524 TI - [Stress during pregnancy and gender of the child]. PMID- 10815526 TI - [Food safety within the EU; the document of the EF committee]. PMID- 10815525 TI - [Regional differences among Danish children's use of antibiotics in 1998]. PMID- 10815527 TI - [Weeks of heart in the years of 2000-2002. Resuscitation--learn yourself to help in cardiac arrest]. PMID- 10815528 TI - [Pericon interactions]. PMID- 10815529 TI - [Medical losses of the Soviet troops during the war in Afghanistan (2)]. AB - Personnel sanitary losses of the 40th Army from gunshot injuries and traumata during the Afghan war took small share in the structure of sanitary losses--about 12%, and fighting sanitary losses constituted 7.49% of the number of all sanitary losses. Significant unevenness in occurrence of fighting sanitary losses was noted--from solitary to 40-50 persons. It allowed to provide their conveyance from field of operations, evacuation to medical institutions and to render the necessary types of medical care in optimal periods. However high frequency of multiple injuries, great share of the casualties with head, abdomen and lower extremity wounds as well as extremely difficult climatic conditions aggravated casualties' state (about 50% needed emergency care). It required revision of organizational-and-staff structure of medical institutions, strengthening of diagnostic units, reanimation and intensive care departments and those units where specialized medical care is rendered, first of all traumatologic and neurosurgical. PMID- 10815530 TI - [Economic problems in military public health]. AB - There are discussed the problems of military treatment and prophylactic institution (TPI) functioning under conditions of market reform of Russian public health. Main marketing concepts in military health are determined and some recommendations on work improvement in TPI of the Armed Forces in the system of obligatory medical insurance are presented, granting population paid medical services. It is necessary to form a new type of director--military and medical manager. PMID- 10815531 TI - [Molecular genetic methods in the practice of military forensic medical expertise]. PMID- 10815532 TI - [Restorative operations on the wounded with colostomies and colonic fistulae]. PMID- 10815533 TI - [Ozone therapy and enteral nutrition in the combined treatment of the enteral failure syndrome in peritonitis]. PMID- 10815534 TI - [The emergency surgery procedure in diseases of the right half of the large intestine]. PMID- 10815535 TI - [The predictors of myocardial electrical instability after its infarction and thrombolytic therapy]. PMID- 10815536 TI - [The use of the methods of R. Foll' and of homeopathy in outpatient practice]. PMID- 10815537 TI - [Current problems in the treatment and prevention of drug-induced toxicodermas]. PMID- 10815538 TI - [The therapeutic activity of Cutivate in the treatment of inflammatory skin diseases]. PMID- 10815539 TI - [The effect of a pulsed magnetic field on the skin microcirculatory bed]. PMID- 10815540 TI - [Russian ultrasonic medical devices]. PMID- 10815541 TI - [The ELMA-1 series electrochemical units for the production of sodium hypochlorite solution]. PMID- 10815542 TI - [The clinical and pharmacodynamic aspects of the antibiotic therapy of infectious patients]. PMID- 10815543 TI - [The improvement in the infectious safety of blood component therapy]. PMID- 10815544 TI - [The dynamics of the physiological indices of servicemen during intermittent alpine adaptation]. AB - Comparative assessment of changes in the main physiologic indices during intermittent (fluctuation) mountain adaptation was conducted in servicemen with various values of physical development and different stability to hypoxic hypoxia. Concrete scientific-and-methodical recommendations on prophylaxis and treatment of dysadaptations which occur during intermittent (fluctuation) mountain adaptation were formulated. PMID- 10815545 TI - [The use of enterosorbents for preventing and stopping chronic toxic stress in submariners]. AB - The efficiency of prophylactic intestinal sorbents use (algisorb and activated carbon) to prevent chronic toxic stress was investigated in 20 healthy volunteers (atomic submarine personnel) basing on evaluation of central nervous system functional state, hemodynamics, hepatic detoxicative ability and dynamics of trace elements in urine. It was revealed that intestinal sorbents administration in daily dose of 5 g during 20 days promotes gastrointestinal elimination of toxic compounds (cadmium, bismuth, chrome, molybdenum, lithium). It is accompanied with decrease in organism intoxication evidences: reduction in central nervous system excessive excitation, decrease in functional tension of cardiovascular system, recovery of hepatic detoxicative ability. Algisorb has an advantage over activated carbon that allows to recommend this drug in prophylaxis and arresting of chronic toxic stress in submariners both during interfield period and autonomic sailing. PMID- 10815546 TI - [Medical support for the combat actions of the frontier units in 1941]. PMID- 10815547 TI - [Medicine in the World War II period in the paintings from the collections of the Military Medical Museum]. PMID- 10815548 TI - [The 80th anniversary of the District Military Clinical Hospital of the Urals Military District]. PMID- 10815549 TI - [The 80th anniversary of the Novosibirsk Military Clinical Hospital]. PMID- 10815550 TI - [The 65th anniversary of the Chelyabinsk Military Hospital]. PMID- 10815551 TI - [The 60th anniversary of the Military Medical Commission of the Northern Fleet]. PMID- 10815553 TI - Laser surgery. Sometimes the right choice, sometimes not. PMID- 10815552 TI - [The 20th anniversary of the Epidemiological Health Detachment of the Northern Fleet]. PMID- 10815554 TI - Health tips. Using household chemicals safely. PMID- 10815555 TI - Old drug gets new life in treating congestive heart failure. PMID- 10815556 TI - HRT may reduce risk of developing colorectal cancer. PMID- 10815557 TI - Aortic stenosis. A murmur from the heart. PMID- 10815558 TI - Breast pain. Mastalgia is common but often manageable. PMID- 10815559 TI - Anxiety. Treatment can help. PMID- 10815560 TI - What's a deviated nasal septum? Does it need to be corrected? PMID- 10815561 TI - How healthy is iceberg lettuce? PMID- 10815562 TI - Silent trauma. AB - Patients who present with discolored and/or symptomatic anterior teeth for which there are no obvious etiologic factors should be carefully questioned regarding their past surgical history. If the patient denies a history of trauma yet relates a recent, (2-5 years) history of undergoing general anesthesia, a differential diagnosis including trauma secondary to endotracheal intubation should be considered. PMID- 10815563 TI - In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media. AB - The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, alpha minimal essential medium (alpha-MEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4 degrees C. A control group was incubated with culture medium at 37 degrees C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2-8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4 degrees C. PMID- 10815564 TI - Experimental comparative study of various mouthguards. AB - The aim of this study was to evaluate the mechanical and physical properties of several standard size commercially available mouthguards. For the purpose of objective testing, a special study model, which the various gum shield devices could be fitted onto, was developed to record tooth deflection caused by impact forces induced by a pendulum ram impact testing machine. The data measured on teeth provided with various gum shield devices were correlated with those of unprotected teeth; this enabled the individual cushioning effects of the respective devices and their specific force conduction to be evaluated. Using mouthguards considerably diminishes the deflection of the teeth subjected to stress in comparison with the row of unprotected teeth. In addition, force is transmitted to the adjacent teeth all the way to the distal regions of the row of teeth. The individual cushioning effects are directly correlated to the thickness of the material; the force distribution is determined by the rigidity of the gum shield device. The devices examined showed considerable differences with regard to force distribution and dimensioning. The study showed that this was due to the thickness of the materials, the manufacturing process and the composition of the materials of the devices examined. In comparison to laboratory-produced devices of similar material thickness, the devices that were designed to be fitted by the user achieved significantly poorer results as regards both cushioning properties and dissipation of exerted forces. PMID- 10815565 TI - Effect of chelating agents on the molecular composition and extent of decalcification at cervical, middle and apical root dentin locations. AB - The aim of this study was to investigate the interaction of two chelating agents used in endodontic treatment with dentin from different root locations. Standardized cervical, middle and apical root dentin specimens were prepared and subjected to treatment with 15% neutral EDTA and RCPrep. Following rinsing with water, dentin surfaces were studied by reflected light optical microscopy and micro-MIR FTIR spectroscopy, while the wash-off extracts were analyzed by atomic absorption spectrometry (AAS) to determine the content of calcium and phosphorus. Neutral EDTA removed the smear layer and opened tubules while RCPrep did not. FTIR measurements showed higher decalcification of dentin surfaces after neutral EDTA treatment although its effect was reduced at apical regions. AAS measurements of calcium and phosphorus confirmed the higher dentin dissolution potential of EDTA at cervical and middle root dentin locations. The results of the present study implied different reaction modes for the two chelating agents tested which might affect the efficiency of the chemo-mechanical preparation when these agents are used. PMID- 10815566 TI - In vitro antibacterial effect of RC-Prep components on Streptococcus sobrinus. AB - The aim of this study was to examine the bacteriostatic and bactericidal effects of the components of RC-Prep: EDTA, urea peroxide and glycol. The minimal inhibitory concentration and the minimal bactericidic concentration of EDTA, urea peroxide and glycol were tested on the viability of Streptococcus sobrinus (S. sobrinus). The combined antibacterial effect of these components was also examined on S. sobrinus. The minimal inhibitory concentration of EDTA was found to be 0.125%, of urea peroxide 0.25%, and of glycol 30%. The minimal bactericidic concentration of EDTA was 0.25%, of urea peroxide 0.5% and of glycol 50%. An antibacterial synergistic effect was found between specific combinations of urea peroxide, EDTA and glycol. No antagonistic effects were found in the various combinations between the components. The antimicrobial effect of the components of RC-Prep was enhanced in specific combinations of the ingredients. This effect might contribute to the intracanal cleansing properties of this medication. PMID- 10815567 TI - Proliferative activity in the juxtaradicular human periodontal ligament. AB - The aim of the present study was to evaluate cell proliferation, assessed by MIB 1, with respect to the type and the distribution of proliferating cells in the healthy juxtaradicular periodontal ligament (PDL) from completely formed human teeth. Immunohistochemical markers against vimentin, CD68 and S-100 were used to characterize cell type. The applicability of the immunohistochemical method on explants of human PDL was also evaluated. The results indicated that under physiological conditions, the majority of the proliferating cells in the PDL were mesenchymal cells predominantly located paravascularly in the middle third of the PDL. Furthermore, MIB 1 reacting with the Ki-67 antigen together with the avidin biotin-complex technique was proved to be an efficient marker of cell proliferation in explants of human PDL. PMID- 10815568 TI - Detection and measurement of endodontic root perforations using a newly designed apex-locating handpiece. AB - Access openings and lateral root perforations were prepared in 40 single-rooted extracted teeth. The distance to the perforations was measured visually and then with the Tri Auto ZX. The electronic device was used first in its apex-locating function and then in conjunction with its automatic reverse mechanism. On average, the electronically measured length was slightly shorter than the visually measured length (P < 0.05). When the automatic reverse mechanism was used, the values registered were either consistent with or slightly different (+/ 0.5 mm) from the electronically measured length (P > 0.05). The results of this study revealed that the Tri Auto ZX detected and measured endodontic root perforations within a range of clinically acceptable variations. PMID- 10815569 TI - Computerized tomography in the management and follow-up of extensive periapical lesion. AB - Radiographs are good diagnostic aids in endodontics, although they have limitations. The purpose of this article is to discuss the use of computerized tomography in differential diagnosis, treatment planning, follow-up and overall clinical management of complex periapical lesions. A clinical case of an extensive symptomatic periapical lesion of the upper jaw is presented, in which the use of computerized tomography allowed evaluation of the true extent of the lesion and its spatial relationship to important anatomical landmarks. Computerized tomography also provided specific information about the type of lesion and the degree of bone repair which had taken place 18 months after non surgical treatment had been completed. PMID- 10815570 TI - Bilateral pulmonary aspiration of intact teeth following maxillofacial trauma. AB - Tooth aspiration is one of the rare sequelae of dental trauma. When this does occur, the right bronchus is usually involved in adults due to the anatomical configuration. In this unusual case, two teeth were aspirated with one entering each lung. While the outcome from cases of aspiration of foreign bodies is usually favourable, this case serves to illustrate one of the possible unfavourable consequences. PMID- 10815571 TI - Child protection: a survey of experience and knowledge within the dental profession of New South Wales, Australia. AB - AIM: To establish the experience and knowledge of dental practitioners in New South Wales, Australia in the area of child abuse and protection. DESIGN: A cross sectional telephone questionnaire. SAMPLE: One hundred and twenty-two interviews; 67 general dental practitioners randomly selected and 55 members of the Australia and New Zealand Society of Paediatric Dentistry. RESULTS: All of the dentists interviewed could identify physical abuse and most identified emotional abuse, however, none could describe all five forms of abuse. Significantly more (58%) members of the paediatric dental society had at some time suspected abuse, compared with only 24% of the general practitioners. However, only 20 of the paediatric dentists and seven general practitioners had actually reported any cases. The main reason given for not reporting concerned confidentiality. Seventy two per cent of general practitioners did not know their legal obligations or with whom they could discuss any concerns. While paediatric dentists achieved a better percentage, still over half of them were similarly unsure. CONCLUSIONS: Further education and training is required for both the dental profession and the child protection agencies to improve the inter-agency working relationship and thus provide better protection for children. PMID- 10815572 TI - The diagnostic value of routine intra-oral premaxillary radiographs in orthodontic assessment. AB - OBJECTIVES: The purpose of this study was to determine whether significant dental abnormalities are likely to be overlooked in the anterior region of the maxilla if, in the absence of any clinical indication for further views, the only radiograph used during an initial orthodontic assessment is a good-quality orthopantomogram (OPT). SAMPLE AND METHODS: Two orthodontists examined, retrospectively, the written and radiographic dental hospital records of 1169 consecutive, new, young patients who on their first visit to a dental teaching hospital underwent radiographic examination comprising an OPT and one or more supplementary radiographs of the anterior maxilla. RESULTS: In five cases (0.43%) significant findings would have been overlooked if the intra-oral views had not been taken. These included periapical lesions and supernumerary teeth, but in three of the cases the image quality of the OPT was poor. CONCLUSIONS: If the OPT is not routinely supplemented by intra-oral views, the chances of completely missing significant findings in the anterior maxilla are small, provided a thorough history and clinical examination have been completed and the image quality of the OPT is good. PMID- 10815573 TI - Association between early weaning, non-nutritive sucking habits and occlusal anomalies in 3-year-old Finnish children. AB - OBJECTIVES, DESIGN AND SUBJECTS: Earlier results on the effect of breastfeeding on the one hand, and non-nutritive sucking habits (pacifier and/or digit sucking) on the other, on the orofacial development of infants are inconclusive. Thus we studied the prevalence of malocclusions and their relationship to the duration of breastfeeding and to non-nutritive sucking habits in a group of randomly selected 3-year-old children (n = 148). RESULTS: Posterior crossbite was detected in 13%, anterior open bite in 18% and large overjet (> 3 mm) in 26% of the children. The proportion of children with anterior vertical open bite was significantly larger among children with non-nutritive sucking habits than among the other children (P < 0.001). Neither posterior crossbite nor large overjet were associated with non nutritive sucking habits. The duration of exclusive breastfeeding was 5.8 +/- 3.6 months while that of total breastfeeding was 7.3 +/- 3.8 months on average. The exclusive and total breastfeeding periods of children with posterior crossbite were both significantly shorter than those of the other children (P < 0.01 and P < 0.002, respectively). Children's height and weight were unrelated to the duration of breastfeeding or occlusal anomalies. CONCLUSION: We suggest that an early introduction of bottlefeeding, indicating a pattern of low-impact muscular activity, may interfere with the normal development of alveolar ridges and hard palate, and hence lead to posterior crossbite. PMID- 10815574 TI - Primary double teeth. A retrospective clinical study of their morphological characteristics and associated anomalies. AB - AIM: To investigate the relationship between morphology and position of double teeth, and the occurrence of other dental anomalies in the same subjects and in their siblings. SAMPLE OF CHILDREN AND METHODS: Fifty-three double teeth in a group of 50 Spanish children were included in the study. All of these children were examined clinically and had radiographs and photographs taken at the time of examination. Twenty-two of the children had a total of 30 siblings who were also examined for the presence of anomalies. RESULTS: Of the 50 subjects, 47 had one and three subjects had two double teeth. Statistically there were no significant differences in occurrence between boys and girls, left and right sides or between maxilla and mandible. Four morphological types were identified: type I, bifid crown-single root; type II, large crown-large root; type III, two fused crowns single root; type IV, two fused crowns-two fused roots. Type I was seen only in the maxilla and types II and III only in the mandible. Type IV was seen mostly in the maxilla. More than half of the cases showed associated anomalies in the permanent dentition and anomalies of number or shape were also present in six of the siblings. CONCLUSION: It is suggested that both double teeth and other anomalies in the same children or in their siblings may be manifestations of a primary abnormality in the distribution of dental material. PMID- 10815575 TI - Microleakage of composite resins in cavities of upper primary molars. AB - OBJECTIVE: To improve the quality of resin restorations, studies are necessary not only of the physical properties of materials but also of the cavity geometry. We have evaluated marginal microleakage for two types of photo-curing composite resin applied to cavities with different marginal structures. SAMPLE AND METHODS: The test materials were 91 extracted upper primary molars. The two composite resins used in this experiment were Liner Bond II (LBII) and Photo Clearfil A (PCA). A class I cavity was created in each tooth; the cavity configuration was a round bevel, a straight bevel and a butt joint. The tooth was then restored with each composite resin. Each tooth was exposed to thermal cycles, followed by immersion in 0.2% basic fuchsin solution. Three sections, through the mesial, central and distal areas of each tooth were taken. The penetration of the dye was examined with a stereoscopic microscope. The sites of observation for examination of microleakage were awarded scores of between 0 and 7. RESULTS: LBII-among the three marginal forms, absence of leakage was observed most frequently in the straight bevel group followed by the round bevel and the butt joint groups, in the mesial and distal sections. In the central section, the straight bevel did not show any scores of 0, 6 or 7. PCA-an absence of leakage in the mesial section occurred most frequently for the round bevel group. However, the absence of leakage in the central and distal sections was most frequent for the straight bevel group. Microleakage of the central section (with scores from 1 to 7) was frequently lower than that for the mesial and distal sections. Scores of 5 and above in the central section were observed only in the butt joint group. The frequency and mean score of leakage were clearly higher in LBII than in PCA, with one significant exception. For both composite resins, the butt joint group showed higher-mean scores of leakage than did either the round or straight bevel group, but for PCA, the mean score of the central section was lower than the other two sections in all groups. CONCLUSION: Our study shows that beveling does reduce marginal leakage. This leakage was more frequently observed with the use of LBII than with PCA. However, the amount of tooth ground away was greater for the bevel treated cavity. In considering these features, special care is necessary in selecting the bur for treating a bevelled cavity. PMID- 10815576 TI - An oral health promotion programme for nursing caries. AB - AIM: To evaluate an oral health promotion programme involving health visitors and mothers of 8-month-old babies in order to address some of the risk factors associated with nursing caries. DESIGN: Two cross-sectional studies using postal questionnaires. SAMPLE: A random sample of 250 mothers who had not received the oral health promotion programme and 250 mothers who had received the programme. SETTING: The City of Salford. RESULTS: The oral health promotion programme significantly improved mothers recall of advice given by health visitors encouraging the use of a feeder cup, brushing their babies' teeth with fluoride toothpaste and restricting sugary foods and drinks. Significant improvements were also found in recall of advice regarding the use of sugar-free medicine and registering babies with a dentist. The programme encouraged a higher proportion of the mothers to bring their children to clinics for a hearing check. CONCLUSION: A simple oral health promotion programme facilitated by health visitors improved recall of advice and clinic visiting behaviour. PMID- 10815577 TI - The relationship between bone age, chronological age and dental age in children with isolated growth hormone deficiency. AB - AIM: To relate dental age and bone age to chronological age in a group of children with isolated growth hormone deficiency (GHD). DESIGN: A group of 25 children between the ages of 6 and 18 years, diagnosed as having GHD were studied. Of these, 16 were male and nine were female, undergoing replacement therapy with growth hormone (GH) over 2.5 years. Dental age (DA) was estimated from orthopantograms (Haavicko Methods) and compared to bone age (BA) by Greulich and Pyle Standards, and chronological age (CA). RESULTS: CA versus BA--there was a statistically significant difference of 1.52 years between the average chronological age and bone age (t = 5.61, P < 0.001). CA versus DA--there was a statistically significant difference of 0.92 years between the average chronological age and dental age (t = 3.93, P < 0.001). BA versus DA- statistically nonsignificant differences were found between the average bone age and dental age (t = -0.60, P = 0.10). CONCLUSION: After 2.5 years of therapy with growth hormone both DA and BA showed a significant delay compared to CA. Despite the fact that some catch-up occurs in speed of growth there is no evidence from the children in our study that might support the occurrence of a similar speed-up in dental age. PMID- 10815578 TI - Witkop tooth and nail syndrome: report of two cases in a family. AB - Tooth and nail syndrome, an autosomal dominant condition, is characterized by hypodontia and nail dysplasia. Mandibular incisors, second molars and maxillary canines are the most frequently missing teeth. Tooth shape may vary and conical and narrow crowns are common. Nail dysplasia affects fingernails and toenails and is often more severe in childhood. Nails may be spoon-shaped, ridged, slow growing and easily broken. The clinical and radiographic features of a father and son presenting with this rare condition are described. PMID- 10815579 TI - Three cases of palatal polyps in infants. AB - Fibrous lesions are common in the oral cavity, however, in infants they are rarely reported. We present three cases of palatal polyps in infants aged 2 days, 3 months and 7 months. In two cases, the treatment was surgical removal and in one case the polyp decreased in size and surgical removal was not required. In two infants, the diagnosis was confirmed histologically as fibroepithelial hyperplasia. PMID- 10815580 TI - Dental findings in three siblings with Morquio's syndrome. AB - Three siblings with Morquo's syndrome are described. Cultured fibroblasts from the youngest sibling demonstrated a total absence of N-acethylgalactosamine-6 sulphate-sulphatase whereas beta-galactocidase activity was normal, thus verifying the diagnosis of MPS-IV A. Dental features such as pointed cusps, spade shaped incisors, thin enamel and pitted buccal surfaces were observed in all three children. Furthermore, in all three siblings the TMJ was affected with severe resorption of the head of the condyle. Histological examination of exfoliated primary molars showed a band of increased porosity following the striae of Retzius in the outer part of the enamel. These developmental disturbances were occasionally associated with minor localized defects in the enamel surface. The importance of close monitoring of dental development and regular dental care in order to prevent attrition of the teeth, loss of vertical face height and subsequent risk of TMJ dysfunction is emphasized. PMID- 10815581 TI - The economic evaluation of pit and fissure sealants. AB - This paper reviews the literature relating to the economic evaluation of sealants. Economic issues are commonly cited for lack of sealant utilization, although there are no adequate economic analyses on which to base this belief. The design of much of the existing economic literature regarding sealants is inappropriate, for example many analyses do not consider the advantages of a sound tooth in the calculation of budget. In general the recent literature supports the efficiency and the effectiveness of sealants. However, there is a need for well designed studies, examining modern materials and populations with current caries prevalence. In particular there is a need for well conducted cost benefit analyses and cost-utility analyses, which should encompass discounting for time and sensitivity analyses. Results from such research would assist clinicians and policy makers alike, in decisions regarding the provision of care with regard to sealant provision at the individual and community level. PMID- 10815582 TI - Prevalence and attitudes to fissure sealants in the general dental service in England. AB - OBJECTIVES: The study objectives were to investigate changes in the prevalence of fissure sealants over a 5-year period among a comparable group of 14-15-year-old regular dental attenders registered with general dental practitioners in 1989 and 1994, and to investigate attitudes towards this form of care amongst general dental practitioners in 1994. METHODS: Random samples of 14-15-year-old pupils were selected from school lists in three areas in England (Doncaster, Hereford/Worcester and Wycombe) in 1989 and 1994. Once parental consent had been gained to examine the children, the Dental Practice Board (DPB) indicated which subjects were 'regular attenders', only these subjects were included in the study. All the subjects in each area were examined by a single trained and calibrated examiner using a standardized technique. In 1994, a group of randomly selected dentists in each area completed a telephone questionnaire assessing attitudes and treatment patterns regarding fissure sealants. RESULTS: In 1989, 2836 letters were posted to parents asking for consent to examine their child and 2174 positive responses were received (response rate 77%). Of the 1919 children examined, 459 were classed as regular attenders, 140 in Doncaster, 159 in Hereford/Worcester and 160 in Wycombe. In 1994, 2560 consent letters were distributed to children in school to pass to their parents and 1587 positive responses were received (response rate 62%). Of the 1388 children examined, 891 were classed as regular attenders, 294 in Doncaster, 318 in Hereford/Worcester and 279 in Wycombe. Large rises in sealant prevalence were observed in all areas, but particularly in Doncaster where sealant prevalence increased from 13% in 1989 to 50% in 1994. In Hereford/Worcester it increased from 25% to 47% and in Wycombe it increased from 16% to 30%. There was also a fall in mean DMFT in all areas. In Doncaster it fell from 2.97 in 1989 to 1.82 in 1994, in Hereford/Worcester it fell from 2.60 to 1.83 and in Wycombe it fell from 1.86 to 1.29. In all three areas the changes in sealant prevalence and DMFT were statistically significant. The questionnaire revealed positive attitudes towards fissure sealants and their usage as well as a number of areas of concern that could potentially hinder sealant usage. CONCLUSIONS: Dramatic increases in sealant prevalence and considerable reductions in dental caries among these regularly attending adolescents have been demonstrated. Some concerns were highlighted regarding sealant usage, which may be prejudicing further increases in their application. PMID- 10815583 TI - Discrimination of morphological findings in dentine from osteogenesis imperfecta patients using combinations of polarized light microscopy, microradiography and scanning electron microscopy. AB - OBJECTIVES: The aim of this study was to investigate the morphological appearance of dentine in teeth from individuals with osteogenesis imperfecta type I, III and IV using different histological techniques, and to correlate morphological findings to different types of osteogenesis imperfecta. SAMPLE AND METHODS: Extracted or exfoliated primary and permanent teeth were collected from 15 patients with the osteogenesis imperfecta diagnoses I, III or IV, with or without the additional diagnosis dentinogenesis imperfecta. Ground and decalcified sections were prepared from the teeth. Histo-morphological studies of the dentine were performed utilizing light and polarized light microscopy, microradiography and scanning electron microscopy. RESULTS: Characteristic findings were irregular tubules, remnants of capillary inclusions and obliterated pulps. All types of osteogenesis imperfecta exhibited similar types of dentine aberrations, but patients with type III or IV had a higher frequency of aberrations when compared to type I. CONCLUSIONS: The combination of either polarized light microscopy or micro-radiography, together with scanning electron microscopy, gave the most amount of morphological information from dentine samples. In addition, aberrations in dentine structure were more clearly observable. Light microscopy was not critical for the analyses. PMID- 10815584 TI - Dental maturation in British children: are Demirjian's standards applicable? AB - OBJECTIVE: The objective of this study was to determine if the standards of dental maturation of Demirjian et al. (1973, 1976) are applicable to British children. DESIGN: The design was cross-sectional, retrospective. SAMPLE AND METHOD: The sample comprised 521 London children of Bangladeshi and white Caucasian (English, Welsh and Scottish) origin aged between 4 and 9 years. Dental age was assessed by crown and root stages of seven mandibular teeth from rotational pantomographs. Dental age was compared to chronological age using a t test. RESULTS: Differences in dental maturation between the two ethnic groups were not significant. British children as a group were dentally advanced compared to the Canadian standards. The mean (+/- standard deviation) advancement in girls was 0.51 +/- 0.79 years and in boys was 0.73 +/- 0.73 years. CONCLUSIONS: The standards of dental maturation described by Demirjian et al. (1973, 1976) may not be suitable for British children. PMID- 10815585 TI - Delayed development or congenital absence of a single first permanent molar in Japanese child patients. AB - OBJECTIVE: Delayed eruption of the first permanent molar (M1) is a relatively rare finding. This report describes the occurrence of delayed development and eruption of a single M1 in Japanese child patients. DESIGN: Cases were identified by surveying 353 orthopantomograms taken of child patients attending a Dental Hospital. Dental age, crown morphology and the presence of third permanent molars (M3) were determined from radiographs and clinical examination. SETTING: Osaka University Dental Hospital in Japan. RESULTS: Nine cases were identified from radiographs. The occlusal morphology of the affected molars showed a reduction of the distolingual cusp. Dental ages correlated more closely to chronological ages when the affected teeth were assumed to be second permanent molars (M2) than they did when the teeth were assumed to be M1. M3 proximate to the affected molar was congenitally absent in the orthopantomograms of all affected subjects. CONCLUSIONS: It is suggested that in the cases seen here the affected teeth were likely to have been second permanent molars that had migrated mesially in association with the congenital absence of M1, although congenital absence of M1 has been reported to be extremely rare. PMID- 10815586 TI - Eruption disturbances of maxillary permanent central incisors associated with anomalous adjacent permanent lateral incisors. AB - Several causes of eruption disturbances in upper permanent central incisors have been described. This report describes six cases in which the failure of maxillary permanent central incisors to erupt appears to be associated with developmental anomalies in the adjacent permanent lateral incisors. The root formation of the unerupted central incisors was delayed in comparison to the contralateral incisors as was the development of the adjacent lateral incisors. The shapes of the crowns were abnormal and in five cases out of the six described, the primary predecessors of the unerupted incisors were fused. PMID- 10815587 TI - A review of 47 cases of unerupted maxillary incisors. AB - OBJECTIVES: To study the prevalence of aetiological factors associated with unerupted maxillary incisors and to follow the outcome of treatment in a study of 47 cases. DESIGN: A retrospective study. SETTING: The Dental Department, St Luke's Hospital, Malta, the School Dental Clinic of Malta and the private practices of two orthodontists in Malta. SAMPLE AND METHODS: Forty-seven patients with a total of 53 unerupted maxillary incisors were classified according to the aetiological factors causing non-eruption. The relative prevalence of the various aetiologies were ascertained, and the outcome after treatment was recorded to assess the efficacy of the treatment methods being used. RESULTS: The most common cause of lack of eruption was the presence of supernumerary teeth (47% of patients). The other 53% of cases were distributed more or less equally between the remaining aetiological factors, which were odontomes (9%), dilacerations (9%), tooth germ malposition (12%), crowding (4%), one case of a calcifying odontogenic cyst (2%) and one case of trauma to the preceding deciduous tooth (2%). The aetiology of 15% of cases could not be ascertained. Once supernumerary teeth were removed, maxillary incisors usually erupted successfully with the help of conventional treatment methods such as surgical exposure and orthodontics. A relatively large number of incisors that failed to erupt due to other aetiological factors had to be extracted. CONCLUSIONS: Maxillary incisors that fail to erupt due to the presence of supernumerary teeth have a better prognosis than unerupted incisors with less common aetiologies. PMID- 10815588 TI - Incontinentia pigmenti: seven cases with dental manifestations. AB - Incontinentia Pigmenti (Bloch-Sulzberger syndrome) is an uncommon genodermatosis that usually affects female infants. The condition is characterized by four cutaneous stages and is frequently associated with dental, ocular, central nervous system and structural anomalies. A large case series of seven patients, all female, who presented to the Department of Paediatric Dentistry at the Eastman Dental Hospital over the last 16 years is reported. The dental features of these cases were typical and included missing teeth, microdontia and delayed eruption. In two of the seven cases, both maxillary canines were palatally impacted. PMID- 10815589 TI - Self-inflicted injury in a case of Hallervorden-Spatz disease. AB - Hallervorden-Spatz disease (HSD) is a rare neurodegenerative disorder characterized by abnormally high deposits of iron in the brain. This report describes a child with HSD who presented with self-inflicted ulceration of the lip and tongue, which was initiated during periods of intense oro-facial spasms. Other findings included dental caries and trauma to the primary incisors. Comprehensive dental care was carried out under general anaesthesia. The self mutilation of the oro-facial mucosa was eliminated by placement of upper and lower soft resin bite guards. PMID- 10815590 TI - Mandibular talon cusp: report of two rare cases. AB - Two rare cases of talon cusps in the mandibular incisors were observed during a prevalence study on talon cusp. One of them was seen on a mandibular primary lateral incisor. The other one was observed on the mandibular left permanent central incisor, and the right mandibular permanent central incisor was congenitally missing. There were no associated developmental syndromes with either of the cases reported. Of the 4770 children examined, nine cases had talon cusp with only two rare cases of mandibular talon cusp, one in the primary dentition and the other in the permanent dentition. PMID- 10815591 TI - UK National Clinical Guidelines in Paediatric Dentistry. Stainless steel preformed crowns for primary molars. Faculty of Dental Surgery, Royal College of Surgeons. PMID- 10815592 TI - Formation of the biologic width following crown lengthening in nonhuman primates. AB - The purpose of this study was to determine if and how the biologic width is reestablished following surgical crown lengthening. Crown-lengthening surgery was performed on the right or left maxillary and mandibular central and lateral incisors of three adult monkeys, with contralateral teeth serving as unoperated controls. Twelve weeks after surgery, tissue blocks were removed for histologic analysis. The results of a histometric evaluation indicate that the biologic width is reestablished following surgical crown lengthening. The junctional epithelium generally migrates to the apical level of root planing. Space for the supracrestal connective tissue fiber groups is created by crestal resorption of alveolar bone. PMID- 10815593 TI - Optimization of resilience and stress distribution in porcelain veneers for the treatment of crown-fractured incisors. AB - The present study was conducted to define, when restoring extensive loss of dentin, the configuration of the restoration that will best reproduce the biomechanical properties of the intact original tooth in terms of resilience and stress distribution. The treatment of 1/3-crown fractures and 2/3-crown fractures was investigated using different designs of facial porcelain veneers with and without underlying composite buildup. The stress distribution and tooth compliance were assessed in a numeric model reproducing a 2-dimensional buccolingual cross section of an incisor. A 50-N facial force was applied to simulate an incisal impact situation. The facial surface tangential stresses were calculated, and the maximum displacement (horizontal direction) at the most incisal node of the enamel surface was also recorded and used to calculate the tooth compliance (i.e., displacement/load or resilience) for each test condition. Tensile stresses were generated on the facial surface of the porcelain laminates with a similar pattern for all test conditions, the cervical part of the crown being the most quiescent area. Substantial differences appeared in the incisal half of the crown, the lowest stresses being observed for extensively fractured teeth restored without composite buildup (facial peaks at approximately 33 MPa). Fractured teeth restored with minimal veneers and a "dentin-like" composite buildup showed stress patterns similar to the intact tooth (facial peaks at approximately 50 MPa). The natural tooth gave the highest tooth compliance or flexibility. All restorative designs featured increased tooth stiffness. However, the original tooth compliance was almost restored when composite was used to replace the missing dentin, with the porcelain acting only as a facial and incisal enamel substitute. When restoring crown-fractured incisors, tooth compliance and stress distribution can be modulated by the combination of composite and ceramics. Optimized configurations can be reached to reproduce the original biomechanical behavior of the intact tooth. The use of ceramic alone generates low stress concentrations, but also less compliant restored teeth. PMID- 10815594 TI - The short-term effect of apically repositioned flap surgery on the composition of the subgingival microbiota. AB - The purpose of this investigation was to examine the short-term effect of apically repositioned flap surgery on clinical and microbiologic parameters in patients with adult periodontitis. A total of 11 patients with moderate to advanced periodontitis received apically repositioned flap surgery. Subjects were monitored during a 3-month pretreatment phase, the baseline surgical phase, and for 3 months post-surgery. Clinical assessments including plaque accumulation, gingival redness, suppuration, bleeding on probing, pocket depth, and attachment level were made at 6 sites per tooth. Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 29 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and percentage of sites colonized by each species (prevalence) were computed for each subject at each visit. After surgery, there was a significant decrease in mean pocket depth and percentage of sites exhibiting gingival redness. Significant decreases were seen in the percentage of sites that had attachment levels < 4 mm, with a significant increase in the percentage of sites with attachment levels of 4 to 6 mm after therapy. The mean total DNA probe count for all bacterial species was significantly decreased by both scaling and root planing and surgical therapy. P gingivalis and B forsythus, 2 bacteria previously shown to be susceptible to mechanical therapy, exhibited statistically significant decreases in mean total DNA probe count. Because surgical therapy decreased levels of the suspected periodontal pathogens C rectus, P nigrescens, and C gracilis, it may be speculated that there was a potential added beneficial effect of surgery on the periodontal microbiota. PMID- 10815595 TI - An introductory therapeutic modality for restoring implants and natural teeth using conus crowns: a 5-year case report. AB - The advent of cylindric titanium implants and the prevalence of periodontal disease have necessitated a restoration that facilitates the performance of oral physiotherapy. The use of the conical crown as a metallic covering over natural teeth and implants--with a suprastructure that is retained through a specific degree of convergence without cement or screws--allows access for enhanced oral hygiene practices, while providing for esthetic and functional goals. PMID- 10815596 TI - Surgical technique for treatment of infrabony defects with enamel matrix derivative (Emdogain): 3 case reports. AB - A surgical protocol is described for the placement of Emdogain enamel matrix derivative during new attachment procedures. Three cases with infrabony defects were treated and a significant probing attachment level (PAL) gain, probing depth reduction, and bone fill were evident on clinical probing and during reentry procedures. The first patient presented a combined one-walled and circumferential defect at a maxillary central incisor. After 1 year the PAL gain was 7 mm. The second case showed a 3-walled defect distal to a maxillary canine. After 1 year the PAL gain was 8 mm, and a reentry procedure showed an almost total fill of the defect. The third patient presented a combined one- and 3-walled defect in the most apical part of the mesial aspect of a maxillary central incisor. One year after the surgical procedure, an orthodontic treatment was performed in this patient. After 6 more months the soft tissue showed a very good esthetic appearance, the papilla height was fully maintained, and there was a PAL gain of 5 mm; 18 months after surgery, reentry showed a significant regeneration of hard tissue that was impossible to probe. Because of these encouraging clinical results, further studies should be initiated to investigate the efficacy of the enamel matrix derivative in new attachment procedures. PMID- 10815597 TI - The simplified papilla preservation flap. A novel surgical approach for the management of soft tissues in regenerative procedures. AB - A novel surgical procedure specifically designed to access interdental spaces in the regenerative treatment of deep intrabony defects is presented. This procedure (simplified papilla preservation flap, SPPF) was designed to provide surgical access to interproximal bony defects while preserving interdental soft tissues, even in narrow interdental spaces and posterior teeth. A modified mattress suture allows coronal positioning of the buccal flap and primary closure of the interdental space without tension. The modified mattress suture minimizes the collapse of the membrane into the defect. An experimental population of 18 patients in good general health who presented with one intrabony defect each was selected for this clinical study. The application of the SPPF in combination with bioresorbable membranes resulted in clinical attachment level (CAL) gains of 4.9 +/- 1.8 mm at 1 year. The difference between baseline CAL and 1 year CAL was highly clinically and statistically significant. The residual pockets at 1 year measured 3.6 +/- 1.2 mm. A slight increase in gingival recession was noted. Primary closure of the flap in the interdental space over the membrane was obtained in 100% of the cases after completion of surgery and maintained in 67% of the cases during the healing period. The application of SPPF in combination with bioresorbable barrier membranes allowed primary closure of the interdental space in most of the treated sites and resulted in consistent CAL gains at 1 year. PMID- 10815598 TI - Clinical and histologic evaluation of the demineralized freeze-dried bone membrane used for ridge augmentation. AB - The biologic principle of guided bone regeneration has been successfully used to regenerate alveolar ridges. The objective of this pilot study was to clinically and histologically evaluate the demineralized freeze-dried bone membrane. Four premolar teeth were extracted from two dogs, and the remaining alveolar bone was surgically reduced in width and height to produce Class III ridge defects. After a 2-month healing period, mucoperiosteal flaps were elevated, and demineralized freeze-dried cortical columns were used as space maintainers. Bone membranes were used as barriers. The animals were sacrificed at 3 months and the surgical areas were recovered and processed for histologic evaluation. Results showed ridge augmentation in all sites. Clinically, the augmented areas appeared to have the same hardness as the surrounding bone on the periphery of the experimental site. After elevation of a mucoperiosteal flap, the bone membrane could be seen. Histologically, the bone membrane acted as an efficient barrier, excluding the nonosteogenic tissues. New bone formation underneath the membrane was found in all specimens. This study suggests that a bone membrane in combination with a space maintainer can guide new bone formation to regenerate localized chronic alveolar ridge defects. PMID- 10815599 TI - Esthetic replacement of a maxillary central incisor with an ITI 15-degree angled implant: a case report. AB - Single-stage implants are frequently used, but their use in the anterior dentition has not been widely published. The purpose of this case report is to present the esthetic result of a single-stage implant to replace a maxillary central incisor, and to demonstrate the unconventional use of a 15-degree angled ITI implant. A 15-degree 12-mm hollow cylinder was placed with the angle reversed facially. The emergence profile of the adjacent roots was matched with the transmucosal portion, turning at the osseous crest. The implant was restored after 6 months with an ITI gold coping and transverse screw-retained crown. The gingival margin and the papillae were shaped by the transmucosal portion and the restoration. To date, the implant has been loaded for 24 months with an ideal soft tissue esthetic and radiographic appearance. PMID- 10815600 TI - A retrospective evaluation of 259 single-tooth replacements by the use of Branemark implants. AB - PURPOSE: The main purpose of this retrospective study was to investigate the clinical outcome of the treatment with 259 consecutively placed Branemark system implant-supported single crowns. MATERIALS AND METHODS: There were 259 implants placed in 183 patients with a mean age of 29.8 years; 230 (89%) were placed in the maxilla and 29 (11%) were placed in the mandible. Of the prosthetic restorations, 165 were all-ceramic (68%), 79 were metal-ceramic (31%), and 4 were gold-acrylic (1%). The observation period ranged from 1 to 9 years. RESULTS: A total of 9 patients (10 implants) was lost to follow-up. There were 3 implant failures registered, one before loading and one during the first year in function; one implant fractured after 6 years in service, giving a cumulative success rate for implants of 98.3%. A total of 8 crowns was remade following prosthetic complications; otherwise, the frequency of adverse events was low. The bone loss was of the same magnitude as in other studies on Branemark implants as support for single crowns. In general, the soft tissue conditions around the restorations were healthy and comparable to those of the patients' natural teeth. CONCLUSION: This study confirms the favorable results presented in other studies on Branemark implants to support single-tooth restorations. PMID- 10815602 TI - Changes in appearance of silicone elastomers for maxillofacial prostheses as a result of aging. AB - PURPOSE: The purpose of this study was to investigate the influence of certain defined variables on color and opacity of silicone elastomers for maxillofacial prostheses. MATERIALS AND METHODS: Three condensation-type and five addition-type silicone elastomers were tested for their changes in color and opacity as a result of aging. The specimens were aged under a xenon light source, dry or wet, and in darkness, also dry (control specimens) or wet. The aging times were 24, 96, 168, 336, 504, 840, 1176, and 1512 hours. The changes in appearance were measured with a spectrophotometer. RESULTS: The condensation-type polymers increased in opacity in an aqueous environment, while the addition-type polymers, as a group, showed the smallest color changes. Although the addition-type polymers generally had a higher filler content than the condensation types, they had a lower opacity. However, because of their higher viscosity, the condensation type polymers offer better possibilities for intrinsic coloring of the prosthesis. CONCLUSION: Under the experimental conditions studied, significant differences between the silicone elastomers regarding color and opacity changes were demonstrated. However, for a proper choice of material in a given case, these factors have to be related to biologic and mechanical properties of the material. PMID- 10815601 TI - Maxillary changes under complete dentures opposing mandibular implant-supported fixed prostheses. AB - PURPOSE: The aim of this study was to determine whether a condition similar to Combination Syndrome occurs in patients rehabilitated with a maxillary complete denture opposing a mandibular implant-supported fixed prosthesis. MATERIALS AND METHODS: Standardized clinical procedures measured fit, occlusal integrity, and bone loss in the anterior maxilla in 11 edentulous subjects meeting these requirements, from the patient pool of the Implant Centre, United Dental Hospital, Sydney. RESULTS: A mean annual loss of 0.17 mm in the anterior maxillary ridge height was not statistically significant (P > 0.05). However, increased pressure in the anterior maxillary ridge during occlusion and loss of posterior occlusal contacts in retruded position were noted on one or both sides in all subjects. CONCLUSION: Loss of posterior occlusion could not be related to anterior maxillary bone loss. However, to maintain the integrity of the prostheses and their supporting structures, it is important to schedule periodic recall appointments for review of the occlusion. PMID- 10815603 TI - Porcelain laminate veneers. A retrospective evaluation after 1 to 10 years of service: Part I--Clinical procedure. AB - PURPOSE: Currently available clinical studies on porcelain laminate veneers seem to support their use as a standard treatment method in modern restorative dentistry. The purpose of this study, which comprises 3 parts, was to evaluate the clinical applicability and the survival probability of porcelain laminate veneers for an observation period of up to 10 years. This first part of the study describes the step-by-step clinical procedure. MATERIALS AND METHODS: The reasons for provision of porcelain laminate veneers were mainly for the treatment of worn anterior teeth, surface enamel defects, intrinsic discolorations, and hypoplasia. As the treatment time for try-in, cementation, and finishing of veneers differs significantly from that needed for conventional restorations, the total time required for veneer treatment was evaluated. RESULTS: A total of 205 porcelain laminate veneers, 117 in the maxilla and 88 in the mandible, was placed in 72 patients (43 females and 29 males). Mean cementation time was 38.7 minutes, which increased significantly to 43.9 minutes if single discolored teeth were being restored. Clinical experience and a familiarity with the strict step-by-step procedure followed throughout the study reduced the mean treatment time from 40.8 minutes during the first 5 years to 36.7 minutes in the following years. CONCLUSION: The porcelain laminate veneer concept provides excellent esthetic and conservative restorations, but it should be stated that the cementation and finishing procedure is highly time consuming. PMID- 10815604 TI - The mandibular speech envelope in subjects with and without incisal tooth wear. AB - PURPOSE: The purpose of this study was to describe a mandibular envelope of motion during speech for 2 subject groups. One subject group had experienced no tooth wear and the other had incisal tooth wear. The speech envelope was to be described with regard to its maximal dimensions in the vertical, anteroposterior, and lateral directions, and its most superior and anterior positions in relation to intercuspal position. MATERIALS AND METHODS: An electromagnetic method of jaw tracking was employed to record mandibular speech movements for 60 subjects whose teeth exhibited no tooth wear and 30 subjects with incisal dental attrition. The test speech exercise contained all of the word sounds that occur in the English language. The test was recorded 3 times, and mean values for parameters were reported. RESULTS: The speech envelope of the tooth wear group was significantly smaller (Student's t test, P < 0.01) in all 3 dimensions than that of the normal group. The tooth wear group speech envelope was also located closer to the intercuspal position in the vertical dimension (P < 0.01) and was not as anteriorly placed (P < 0.01). In the normal group, correlations were found between the vertical incisor overlap and the most superior speech position (r = 0.48, P < 0.0001), and between the horizontal incisor overlap and the most anterior mandibular speaking position (r = 0.63, P < 0.0001). CONCLUSION: The envelope of mandibular movement during speech differed in dimension and position between the 2 groups investigated. PMID- 10815605 TI - A 10-year longitudinal study of fixed prosthodontics: clinical characteristics and outcome of single-unit metal-ceramic crowns. AB - PURPOSE: The purpose of this study was to report on the clinical characteristics and outcome of 688 single-unit metal-ceramic crowns placed in a specialist prosthodontic practice between January 1984 and December 1992. MATERIALS AND METHODS: Clinical and laboratory techniques were standardized and patients were recalled by the author between June and December of 1993. The outcome of all crowns was allocated to one of 6 fields in this assessment period. RESULTS: Clinical review examination by the author covered 87% of the crowns. Of these, 52% had been in service for 5 to 10 years and 48% for less than 5 years but more than 1 year; 67% of the crowns were placed on vital teeth and 94% were given a "good prognosis" rating at examination. The repair and failure rates of crowns in the 5 to 10 years in clinical service group were both 3%. Crowns on nonvital teeth in the same grouping had a significantly greater failure rate (5%) than crowns on vital teeth (1%), and anterior teeth had a significantly greater retreatment rate than posterior teeth. Retreatment for all 25 crowns occurred within 66 months after cementation. Tooth fracture accounted for 56% of retreatments, while caries and loss of retention accounted for only 24% of retreatments; 2% of initially vital teeth were endodontically treated during the survey period. CONCLUSION: Single-unit metal-ceramic crowns have a high expectancy of exceeding 10 years of clinical service when the described clinical and laboratory protocol is applied. PMID- 10815606 TI - Prosthodontic decision making among general dentists in Sweden. II: The choice between fixed and removable partial dentures. AB - PURPOSE: The purpose of this study was to describe how dentists evaluated various items related to a treatment choice between fixed partial dentures (FPD) and removable partial dentures (RPD), and to determine if the differences could be explained by dentist-related variables ("social and demographic attributes," "job situation," and "attitudes"). MATERIALS AND METHODS: Questionnaires were sent to a random sample of 2,059 Swedish general dentists, with a response rate of 76%. In the questionnaire, the choice between FPDs and RPDs in a clinical situation was presented. The dentists were asked to mark on 14-item visual analogue scales the relative importance he or she gave the different items. The items were analyzed through principal components analysis, where a 3-factor solution was obtained; the factors were labeled as "time," "health," and "comfort." The factors were run as dependent variables in multiple regression analyses. RESULTS: Great individual variations were seen, but the differences between groups of dentists were small. The items evaluated as most important were "patient's wish," "condition of possible abutment teeth," and "prognosis for delivered treatment." Male dentists gave significantly greater importance to the "health" factor compared to female dentists. The attitudinal variable "patient information" showed significant associations with all 3 factors in the multivariate models. CONCLUSION: Great individual differences were seen regarding the importance of the various items. In multiple regression models, several independent variables showed significant associations, most interestingly the attitudinal variable "patient information." Low explanatory (R2) values indicate that it is necessary to capture more variables of importance for the prosthodontic decision-making process. PMID- 10815607 TI - Comparative mechanical property characterization of 3 all-ceramic core materials. AB - PURPOSE: The biaxial flexural strength and fracture toughness for 3 representative types of ceramic core materials were studied to (1) ascertain the ranking of the 3 ceramic types for strength and toughness, and (2) provide clinicians with more information to predict clinical outcomes. The former aim was deemed important in view of the importance of flaw size in the relationship between failure stress and fracture toughness of brittle materials. MATERIALS AND METHODS: The 3 representative ceramic types included a leucite-reinforced glass, a glass-infiltrated sintered alumina, and a high-purity, high-density alumina. The biaxial flexural strength was measured with the piston-on-3-ball method. The plane-strain fracture toughness was measured with the short-bar chevron-notch method. RESULTS: The biaxial flexural strengths of the high-purity, high-density alumina and glass-infiltrated sintered alumina ceramic core types were significantly higher than that of the leucite-reinforced glass ceramic type. The glass-infiltrated sintered alumina was significantly higher in fracture toughness than the high-purity, high-density alumina type, which was significantly higher than the leucite-reinforced glass. CONCLUSION: All materials surpassed the minimum strength requirement outlined by the International Standards Organization, and they also had similar strength variability according to their Weibull moduli. The glass-infiltrated sintered alumina and the high-purity, high density alumina types were significantly stronger and tougher than currently used core materials. However, while the glass-infiltrated sintered alumina had a higher fracture toughness than the high-purity, high-density alumina, it had a lower flexural strength. PMID- 10815608 TI - The effect of chopped poly(methyl methacrylate) fibers on some properties of acrylic resin denture base material. AB - PURPOSE: The fracture of acrylic resin dentures remains an unresolved problem. Over the years, various approaches to strengthening acrylic resin have been suggested, including modifying or reinforcing the resin. The aim of this study was to investigate the effect of chopped poly(methyl methacrylate) (PMMA) fibers on some properties of acrylic resin denture base material. MATERIALS AND METHODS: PMMA in the form of fibers 0.75 mm in diameter and 5 mm in length was added to acrylic resin denture base material in various percentages to form a composite material. The influence on doughing and manipulation times and transverse strength was examined. The results were subjected to statistical analysis using a one-way analysis of variance and, where appropriate, the Scheffe test. RESULTS: The results showed that the doughing time was decreased by the addition of fibers, with the manipulation and setting times showing inconsistent changes. There was a significant difference between the materials in terms of the transverse strength. When the amount of PMMA fibers in the acrylic resin was increased, there was a decrease in the modulus of rupture and a decrease in the modulus of elasticity. The differences were shown to be statistically significant in some groups. CONCLUSION: The doughing time was decreased by the addition of fibers, while the manipulation and setting times showed inconsistent changes. The incorporation of chopped, randomly oriented PMMA fibers into acrylic resin had no advantage over the unmodified polymer in terms of strength and cannot be recommended as a reinforcing agent for acrylic resin denture base material. PMID- 10815609 TI - New approach for evaluating metal-porcelain interfacial bonding. AB - PURPOSE: The purpose of this study was to evaluate the bonding characteristics of porcelain-fused-to-metal (PFM) systems by determining the strain energy release rate associated with interface fracture of porcelain and metals. MATERIALS AND METHODS: Porcelain-veneered metal plates cast from commercially pure titanium and 3 metal alloys (gold, palladium, and nickel-chromium alloys) were made to dimensions of 25 mm x 8 mm x 2.5 mm with comparable thicknesses of porcelain and metal. The porcelain side of the specimens was notched to the interface with a thin diamond saw, and a small precrack was initiated at the metal-porcelain interface. The samples were subjected to a limited number (typically less than 4) of load-unload cycles under 4-point bending at a crosshead speed of 0.1 mm/min. The loading and unloading force displacements associated with stable crack extension were recorded. The strain energy release rate was calculated. The interfacial area was also examined under scanning electron microscope (SEM) after the test. RESULTS: The mean strain energy release rates were 72.7 +/- 10.0 J/m2, 58.5 +/- 13.5 J/m2, 39.4 +/- 4.3 J/m2, and 16.6 +/- 2.5 J/m2 for the samples of gold, palladium, nickel-chromium alloys, and titanium, respectively. The SEM photographs showed that the crack occurred in the porcelain layer close to the interface. CONCLUSION: The bonding characteristics of PFM systems were determined with 3 types of metal alloys and commercially pure titanium by a fracture mechanics approach. The gold alloy and titanium are considered to obtain the greatest and least adhesion, respectively. The test system has proven to be a simple and reliable approach to determine the bonding in biomaterial systems. PMID- 10815616 TI - Serum levels of IL-6 type cytokines and soluble IL-6 receptors in active B-cell chronic lymphocytic leukemia and in cladribine induced remission. AB - We have investigated the serum concentrations of interleukin-6 (IL-6) and two IL 6 family cytokines-oncostatin M (OSM) and leukemia inhibitory factor (LIF)-in 63 patients with B-cell chronic lymphocytic leukemia (B-CLL) and 17 healthy controls using the enzyme-linked immunosorbent assay (ELISA) method. Simultaneously, we measured the serum levels of the soluble forms of two subunits of the IL-6 receptor complex-ligand binding glycoprotein 80 (sIL-6R) and glycoprotein 130 (sgp130). The cytokines and receptors were evaluated in 25 untreated patients and 38 patients treated with cladribine (2-CdA), as well as in 17 healthy controls. We have correlated the serum levels of these proteins with Rai's clinical stage of the disease, the response to 2-CdA treatment and some hematological parameters. We have also evaluated the correlation of the IL-6 serum level with the concentration of OSM and IL-6 soluble receptors. IL-6 was measurable in 62/63 (98.4%), OSM in 20/25 (80%) of untreated and 14/38 (37.8%) of the treated patients. sIL-6R and sgp130 were detectable in all 63 patients and LIF in none of the CLL patients. IL-6 serum level in untreated patients was not significantly different as compared to its concentration in the control group (P>0.05). However, in the patients treated with 2-CdA the IL-6 level was significantly lower (P<0.02), and the lowest concentration was found in the patients with complete remission (CR; median 1.4pg/ml; P<0.02). The concentration of sIL-6R was significantly higher in untreated (median 61.8 ng/ml) and treated (median 50.1 ng/ml) CLL patients when compared to normal persons (median 41.2 ng/ml; P=0.04; P<0.001, respectively). There was no difference between the sIL-6R levels in the patients with CR and the healthy controls. In non-responders sIL-6R concentration was the highest and similar to its level in the untreated patients. OSM level was higher in the untreated patients (median 1.8pg/ml) than in the normal controls (median 0.0pg/ml; P<0.001) and in the CR patients (median 0.0pg/ml; P<0.03). The serum concentration of sgp130 was similar in the untreated (median 480 pg/ml) and treated (median 470 pg/ml) patients, as well as in the healthy persons (median 420 pg/ml; P>0.05). We have found significant positive correlation between the levels of sIL-6R and the lymphocytes count in CLL patients (p=0.423; P<0.001). In addition, sIL-6R and OSM serum concentrations correlated also with CLL Rai stage. In conclusion, the serum level of IL-6, OSM and sIL-6R, but not LIF and sgp130, are useful indicators of CLL activity. PMID- 10815617 TI - The induction of cyclooxygenase-2 in IL-1beta-treated endothelial cells is inhibited by prostaglandin E2 through cAMP. AB - Prostaglandins (PGs) have numerous cardiovascular and inflammatory effects. Cyclooxygenase (COX), which exists as COX-1 and COX-2 isoforms, is the first enzyme in the pathway in which arachidonic acid is converted to PGs. Prostaglandin E2 (PGE2) exerts a variety of biological activities for the maintenance of local homeostasis in the body. Elucidation of PGE2 involvement in the signalling molecules such as COX could lead to potential therapeutic interventions. Here, we have investigated the effects of PGE2 on the induction of COX-2 in human umbilical vein endothelial cells (HUVEC) treated with interleukin 1beta (IL-1beta 1 ng/ml). COX activity was measured by the production of 6-keto PGF1alpha, PGE2, PGF2alpha and thromboxane B2 (TXB2) in the presence of exogenous arachidonic acids (10 microM for 10 min) using enzyme immunoassay (EIA). COX-1 and COX-2 protein was measured by immunoblotting using specific antibody. Untreated HUVEC contained only COX-1 protein while IL-1beta treated HUVEC contained COX-1 and COX-2 protein. PGE2 (3 microM for 24h) did not affect on COX activity and protein in untreated HUVEC. Interestingly, PGE2 (3 microM for 24h) can inhibit COX-2 protein, but not COX-1 protein, expressed in HUVEC treated with IL-1beta. This inhibition was reversed by coincubation with forskolin (100 microM). The increased COX activity in HUVEC treated with IL-1beta was also inhibited by PGE2 (0.03, 0.3 and 3 microM for 24h) in a dose-dependent manner. Similarly, forskolin (10, 50 or 100 microM) can also reverse the inhibition of PGE2 on increased COX activity in IL-1beta treated HUVEC. The results suggested that (i) PGE2 can initiate negative feedback regulation in the induction of COX-2 elicited by IL-1beta in endothelial cells, (ii) the inhibition of PGE2 on COX-2 protein and activity in IL-1beta treated HUVEC is mediated by cAMP and (iii) the therapeutic use of PGE2 in the condition which COX-2 has been involved may have different roles. PMID- 10815618 TI - Prostaglandin E2 affects differently the release of inflammatory mediators from resident macrophages by LPS and muramyl tripeptides. AB - LPS and MTP-PE (liposome-encapsulated N-acetyl-muramyl-L-alanyl-D-isoglutaminyl-L alanine-2-:[1',2'dipalmitoyl -sni-glycero-3-(hydroxy-phosphoryl-oxyl)] etylamide) induce in liver macrophages a synthesis and release of TNF-alpha, nitric oxide and prostanoids. Both agents induce an expression of mRNA's encoding TNF-alpha, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, and of corresponding proteins. LPS and MTP-PE induce a rapid activation of the extracellular regulated kinase (ERK) isoenzymes-1 and -2. Inhibition of map kinase isoenzymes leads to a decreased release of TNF-alpha, nitric oxide and prostaglandin (PG) E2 after both agents. The transcription factors NF-kappaB and AP-1 are strongly activated by LPS within 30 minutes. MTP-PE induces a weak activation of both transcription factors only after 5 hours. Inhibition of NF kappaB inhibits the LPS- but not the MTP-PE-induced release of TNF-alpha, nitric oxide and PGE2. PGE2 release after LPS is higher than after MTP-PE. Exogenously added PGE2 inhibits the activation of map kinase and TNF-alpha release by LPS, but not by MTP-PE. Release of nitric oxide after LPS and MTP-PE is enhanced after prior addition of PGE2. PGD2 is without any effect. MTP-PE, but not LPS, induces a cytotoxicity of Kupffer cells against P815 tumor target cells. The MTP-PE induced cytotoxicity is reduced by TNF-alpha neutralizing antibodies, indicating the involvement of TNF-alpha. Thus our results suggest that the different potencies of LPS and MTP-PE as immunomodulators probably result from different actions on Kupffer cells, resulting in differences in the amounts and kinetics of released TNF-alpha and PGE2, and that PGE2 plays an important regulatory role in the action of LPS, but not in the actions of MTP-PE. PMID- 10815619 TI - Circulating TCR gammadelta cells in the patients with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a disorder with a wide range of immunological abnormalities. The results of the studies undertaken in the last decade indicated that SLE pathogenesis was mainly connected with the breakdown of the activation control of B and T cells, generating humoral or cell-mediated responses against several self-antigens of affected cells. The last studies demonstrate that the role of gammadelta T lymphocytes in autoimmune diseases can be especially important. Flow cytometry techniques were used to investigate the number and percentage of TCR gammadelta T cells and their most frequent subtypes in peripheral blood of 32 patients with SLE and 16 healthy volunteers. We also correlated TCR gammadelta cells number with the level of T CD3+, T CD4+, T CD8+, and NK (CD16) cells (cytometric measurements) and SLE activity (on the basis of clinical investigations). Our studies were preliminary attempts to evaluate the role of that minor T cell subpopulation in SLE. Absolute numbers of cells expressing gammadelta TCR in most SLE blood specimens were significantly lower than in the control group (P<0.006). However, since the level of total T cell population was also decreased in the case of SLE, the mean values of the percentage gammadelta T cells of pan T lymphocytes were almost the same in both analysed populations (7.1% vs 6.3%, respectively). In contrast to Vdelta2+ and Vgamma9+ subtypes of pan gammadelta T cells, Vdelta3+ T cells number was higher in SLE patients (20 x 10 cells/microl) than in healthy control group (2 x 2 cells/microl) (P=0.001). However, we found no differences between the numbers of pan gammadelta T lymphocytes and studied their subtypes in the patients with active and inactive disease. These cell subpopulations were doubled in the treated patients with immunosuppressive agents in comparison with untreated ones; however, data were not statistically significant. Our study indicated that Vdelta3+ subtype of gammadelta T cells seems to be involved in SLE pathogenesis; however, we accept the idea that the autoimmunity does not develop from a single abnormality, but rather from a number of different events. PMID- 10815620 TI - Killer-cell inhibitory receptors, CD158a/b, are upregulated by interleukin-2, but not interferon-gamma or interleukin-4. AB - Although it is now accepted that killer-cell inhibitory receptors (KIRs), which were molecularly cloned in 1995, deliver negative signals to natural killer (NK) cells regarding the recognition of target cells, it is still unclear how the expression of these receptors on lymphocytes is regulated. Therefore, we investigated the regulation of expression of representative KIRs, CD158a and CD158b, by cytokines such as interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN gamma). Neither IL-4 nor IFN-gamma affected the expression of CD158a/b, but incubation for 48 h with IL-2, which enhances the killer activity of NK cells, upregulated the expression of the KIRs. This upregulation by IL-2 was also observed in CD16-positive cells sorted from total lymphocytes. In contrast, IL-4, which is a down-regulator of IL-2-induced killer responses, did not change the level of CD158a/b expression when added after the IL-2 treatment. These findings suggest that IL-2 plays an important role in the regulation of CD158a/b expression, and might be involved in controlling NK activity via regulating expression of these molecules. PMID- 10815621 TI - Basal secretion of lysozyme from human airways in vitro. AB - The aim of this study was to examine the basal release of lysozyme from isolated human lung tissues. Measurements of lysozyme in the fluids derived from lung preparations were performed using a rate-of-lysis assay subsequent to acidification of the biological samples. Lysozyme released from bronchial preparations into fluids was greater than that observed for parenchymal tissues. The lysozyme quantities detected in bronchial fluids were not modified by removal of the surface epithelium. Furthermore, the quantities of lysozyme in bronchial fluids was correlated with the size of the bronchial preparations. These results suggest that the lysozyme was principally secreted by the human bronchi (submucosal layer) rather than by parenchyma tissues and that a greater release was observed in the proximal airways. PMID- 10815622 TI - Radiation-induced lung damage: a challenge for radiation biology, experimental and clinical radiotherapy. PMID- 10815623 TI - The pig as a model for the study of radiation effects on the lung. AB - PURPOSE: The treatment of thoracic malignancies is frequently limited by the 'tolerance' of normal lung tissue. In order to learn more about the factors that influence lung tolerance an animal model that closely mimics the clinical exposure situation is required. The lungs of pigs are similar to those of man in a variety of ways and the animal's size permits the irradiation of partial tissue volumes comparable with those used clinically; very rarely in man is the whole lung irradiated. In this report, the available data for the effects of irradiation on pig lung are reviewed as they relate to the key issues in radiotherapy. RESULTS: The dose-effect relationships for exposure to single doses indicate that for a significant impairment in both early and late lung function and for the histological detection of fibrosis, the dose-related changes in pig and man are similar. Studies with dose-fractionation using X-rays indicate a large dependence of the iso-effective dose on fraction number and fraction size, and the parameters obtained were not significantly influenced by the time of assessment after irradiation. A simple power-law function fitted the whole data set better than the linear-quadratic model, with a fraction number exponent (N) of 0.44+/-0.06 for treatments given in 1-30 fractions. The alpha/beta values ranged from 0.6 to 4.86 Gy, tending to increase with the length of the follow-up period; however; the majority of these alpha/beta values were not significantly different from zero at the 5% level. Studies of the effect of changes in the volume of lung tissue irradiated indicated the need for care in the use of the terms 'tolerance' and 'iso-effective' dose. Doses that were iso-effective for the severity of regional damage were not matched by those for total lung function. The same level of damage in a small volume compared with a large volume had less effect, i.e. was better tolerated in terms of changes in total lung function. CONCLUSION: Iso-effective doses in pig and humans are lower than those for the more common laboratory animal species. This observation may be related to the differences in anatomical structure of the lungs in the different species. PMID- 10815624 TI - Quantitative clinical radiobiology of early and late lung reactions. AB - PURPOSE: To quantify the response of human lung to a course of fractionated radiotherapy based on a literature review of published clinical data. MATERIALS AND METHODS: Quantitative clinical radiobiology is concerned with the estimation of parameters that describe the clinical outcome of radiotherapy as a function of patient and treatment characteristics. Here, parameters describing the steepness of the dose-response curve, the response to a change in dose per fraction and to a change in overall treatment time for early and late lung injury are compiled based on published clinical studies. RESULTS: Two phases of lung injury are seen, radiation pneumonitis and lung fibrosis. The first signs of early lung changes are seen almost immediately after irradiation. This reaction peaks after 5 to 6 months, and settles partially before 9-10 months. Around that time, the late changes become manifest and these are stable in most cases. There is an important distinction between lung injury and radiotherapy-related morbidity, as even severe changes in a small volume may not give rise to any clinical symptoms. Many assays have been developed for lung damage, and these highlight various clinical and biological aspects of lung damage. Here, the literature on steepness of dose response curves and fractionation sensitivity is reviewed and quantified by the alpha/beta ratio of the linear-quadratic model for both radiation pneumonitis and lung fibrosis. For the early phase a significant time factor exists. Current best estimates for these radiobiological parameters are derived. Other external factors affecting these estimates are briefly discussed. CONCLUSIONS: Quantitative estimates of radiobiological characteristics of human lung are available for the pneumonitis phase where the fractionation sensitivity is in the same range as for most late-responding normal tissues. Short intensive schedules may also bear an added risk for pneumonitis as the dose recovered per day is around 0.5 Gy. For the later phase of lung fibrosis, the estimates are fewer and generally less precise. It is clear though, that the alpha/beta ratio is low, possibly 2-3 Gy. No time factor has been demonstrated for the late reaction. Due to the considerable physiological reserve capacity in the normal human lung, the relationship between damage and morbidity depends strongly on the lung volume affected. It therefore seems likely that for small volumes irradiated to high doses, the dose-limiting complications may not be due to restriction of lung function, but rather to haemorrhage and formation of fistulae. PMID- 10815625 TI - Volume and dose-response effects for severe symptomatic pneumonitis after fractionated irradiation of canine lung. AB - PURPOSE: To study the dose-related incidence of severe symptomatic pneumonitis following fractionated irradiation applied to three different volumes of lung in normal beagle dogs. MATERIALS AND METHODS: A three-dimensional treatment planning system was used to design mediastinal fields of increasing width to irradiate 33%, 67% or 100% of both lungs combined in 128 normal beagle dogs. Total doses, ranging from 27 to 72 Gy, were delivered in 1.5 Gy fractions over 6 weeks. RESULTS: No dogs irradiated to 33% of their total lung volume developed severe symptomatic pneumonitis. In the 67% volume group, logistic fit of the data showed a dose-response curve with a 50% probability of developing severe symptomatic pneumonitis (ED50) after a total dose of 56.0 Gy (52.2-66.0 Gy, 95% confidence interval, CI). The more clinically relevant ED5 for the first 6 months after irradiation of 67% of the lung was 48.1 Gy (18.5-52.0 Gy, 95% CI). The ED50 and ED5 values after irradiation of the whole lung (100%) were 44.1 Gy (41.2-53.5Gy, 95% CI) and 39.1 Gy (8.8-41.8 Gy, 95% CI) respectively. CONCLUSION: Severe symptomatic pneumonitis proved to be a very informative volume-effect endpoint, clearly demonstrating that irradiated lung volume is a critical parameter to be considered in assigning thoracic radiotherapy treatment parameters. Volume effects in lung are dependent on the compensatory capacity of the nonirradiated lung. Underlying pathophysiology of irradiated tissue, as well as decreased compensatory capacity of nonirradiated tissue may have a strong effect on the dose-volume response. PMID- 10815626 TI - Radiation-induced pulmonary injury: symptomatic versus subclinical endpoints. AB - PURPOSE: To assess the relationship between radiation (RT)-induced pulmonary symptoms and subclinical changes in pulmonary functions tests (PFT) and radiographs. MATERIALS AND METHODS: A total of 184 patients irradiated between 1992 and 1998 were prospectively evaluated for RT-induced pulmonary symptoms, changes in computed tomography (CT) density, reductions in single photon emission CT (SPECT) perfusion, and changes in pulmonary functions tests (forced expiratory volume in 1 s [FEV1] and diffusion capacity to carbon monoxide [DLCO]). Comparisons between the evaluable patients with (N=34) and without (N=106) RT induced pulmonary symptoms were made. RESULTS: Within 6 months of RT, 80% of the RT-induced symptoms were noted. There was no association between the presence or absence of RT-induced pulmonary symptoms and the frequency of RT-induced radiographic changes (p=0.53), or in the dose-response curve for RT-induced reductions in regional perfusion. Overall, RT-induced changes in SPECT images were more commonly seen than increased density changes on CT (p<0.001). Most patients with pulmonary symptoms had relatively low pre-RT PFTs and experienced further declines following RT. CONCLUSIONS: Regional radiographic changes in CT defined tissue density or SPECT-defined tissue perfusion are similar in patients with and without RT-induced pulmonary symptoms because these endpoints do not consider the volume of lung affected. RT-induced pulmonary symptoms are better related to post-RT PFT because they are an assessment of whole lung function. Additional studies are necessary to better define models that can predict the degree of radiation-induced changes in whole lung function. PMID- 10815627 TI - Lack of effect of small high-dose volumes on the dose-response relationship for the development of fibrosis in distant parts of the ipsilateral lung in mini pigs. AB - PURPOSE: Multi-field radiation therapy for intrathoracic tumours results in a heterogeneous dose distribution in lung tissue. This study investigated whether irradiation of small lung volumes with high fibrogenic doses affects the dose response relationship for development of fibrosis in distant parts of the ipsilateral lung of mini-pigs. MATERIALS AND METHODS: The whole right lung of 26 'Mini-Lewe' pigs was irradiated with homogeneous doses of between 25 Gy and 40 Gy given in five equal fractions using opposing anterior-posterior portals and a linear accelerator. Another 32 animals were irradiated with a constant dose of 35 Gy to a small house-shaped high-dose field (base 3.0 cm, height 4 cm) located 3 cm caudolateral to the right hilus, while the surrounding right lung received either no irradiation or homogeneous doses of between 20 Gy and 30 Gy. The radiation fields were simulated and port films were obtained for each of the 10 fields in all pigs. Fibrosis was quantified 9 months after irradiation by determination of the hydroxyproline (HP) content of the 32 high-dose volumes and in the lung apex and the basolateral lung of all 58 pigs. Based on the reference value for the HP-ratio, i.e. the HP-concentration of the right lung over the left lung, obtained in 12 unirradiated control animals, the experimental results were converted into quantal data for probit analysis, a responder being an animal with an HP-ratio > 1.33. RESULTS: A dose-response relationship for the HP-ratio was obtained in the different lung sites and irradiation groups. For a given dose level the mean HP-ratios and response rates did not differ systematically between the lung apex and the basolateral lung. Probit analysis of the pooled data produced ED50 values of 21.8 Gy (95% CI 12-37) for irradiation without a high dose volume and 25.9 Gy (24-28) for irradiation with a high-dose volume. These values are not significantly different. The results from both irradiation groups could be well fitted by a common dose-response curve with an ED50 value of 26.1 Gy. Unexpectedly, the response rates in the high-dose volume increased with increasing dose to the surrounding right lung. Analysis of the port films provided an explanation for this finding: inaccuracies in daily field positioning. When this error was corrected for by use of the mean dose to the high-dose volume, a dose-response curve with an ED50 of 25.2 Gy (22-29) was determined for the high-dose volume. CONCLUSIONS: The results of the study indicate that the irradiation of a small lung volume with high fibrogenic doses does not affect the dose-response relationship for development of fibrosis in distant parts of the ipsilateral lung. PMID- 10815628 TI - Down-regulation of SP1 DNA binding activity in the process of radiation-induced pulmonary fibrosis. AB - PURPOSE: To determine changes in the expression and function of the transcription factor SP1 in radiation-induced pulmonary fibrosis. MATERIALS AND METHODS: The right lungs of female Fischer rats were irradiated with a fibrogenic single dose of 20 Gy gamma-irradiation. SP1 mRNA and protein expression was determined by Northern and Western blotting, respectively, between 30 min and 12 weeks after irradiation. Cellular localization of SP1 protein was characterized by immunohistochemistry (peroxidase labelling). SP1 DNA binding activity was studied with electrophoretic mobility shift assays (EMSA). RESULTS: Eight weeks after irradiation, pulmonary fibrosis was first observed. SP1 DNA binding activity showed a short-term increase from 30 min to 12 h after irradiation. Thereafter it remained quite stable until 1 month after irradiation. However, 2 months after irradiation, SP1 DNA binding activity was no longer detectable. The SP1 mRNA level was not reduced at this time, nor was there a reduction in its size. However, Western blotting revealed the occurrence of at least two slightly smaller additional bands 2 months after irradiation whereas the original SP1 band vanished. This suggests a degradation event of SP1 taking place near one or both ends of the protein. Most of the SP1 protein was found in type II pneumocytes and alveolar macrophages of the normal and fibrotic lung. Bronchial epithelial cells were also positive. In the fibrotic lung, proliferating fibroblasts also become positive. CONCLUSIONS: The functional knockout of the transcription factor SP1, in the process of irradiation-induced pulmonary fibrosis, is demonstrated. This should help elucidate the severe disturbances in transcriptional regulation, cellular proliferation and differentiation occurring in the lung at long intervals after irradiation. PMID- 10815629 TI - Immunohistochemical evidence for the occurrence of similar epithelial phenotypes during lung development and radiation-induced fibrogenesis. AB - PURPOSE: Processes of fibrosis, wound healing and tissue regeneration have in common the fact that proliferation and differentiation of cells involved in the restoration of normal-tissue architecture resemble to a certain degree the embryonic development of the corresponding tissue. The present review focuses on the phenotypic changes of alveolar epithelial cells during fibrogenesis and describes similarities in the expression pattern of epithelial antigens during lung development. METHODS: For comparative studies, immunohistochemical investigations of different experimental fibrosis models were performed. RESULTS: For several epithelial proteins, such as the CD44 adhesion molecule, the enzymes carbanhydrase II and cathepsin D, as well as the lectin galectin-3, a transient epithelial immunoreactivity have been detected. What all four examples have in common is that a part of the foetal antigenic profile reappears under conditions of injury and during the development of pulmonary fibrosis. CONCLUSIONS: The re expression of foetal antigens in fibrotic samples with a spatio-temporal pattern, as detected by immunocytochemical techniques, indicates that some mechanisms or factors exist, which similarly regulate the differentiation of the epithelium during ontogenesis and in the remodelling process during fibrogenesis. PMID- 10815630 TI - TGF-beta1-mediated alterations of rat lung fibroblast differentiation resulting in the radiation-induced fibrotic phenotype. AB - PURPOSE: To study the influence of TGF-beta1 and TGF-beta-neutralizing antibodies on the clonogenic activity and terminal differentiation of rat lung fibroblasts following radiation exposure. MATERIAL AND METHODS: Early passage rat lung fibroblasts were used in this study. Colony formation assays were applied to determine the radiation sensitivity as well as radiation-induced alterations in differentiation pattern. Based on a TGF-beta1-specific ELISA system, the amount of TGF-beta1 in the culture medium of sham-irradiated and irradiated cultures was determined. RESULTS: Applying immediate (ip) and delayed plating (dp) procedures for cells irradiated in the subconfluent state, distinct differences in the radiation dose-response curves could be observed (SF2ip 0.20+/-0.025 versus SF2dp 0.51+/-0.0077). Upon irradiation with a single dose of 4Gy the level of TGF-beta1 found in the culture medium increased by about 60%. Radiation-induced terminal differentiation of progenitor fibroblasts (MF) to postmitotic fibrocytes (PMF) was expressed by the change of the ratio of PMF:MF (0.8 at 0 Gy versus 3.0 at 4 Gy). Neutralizing antibodies directed against TGF-beta inhibited both the radiation-induced reduction in clonogenic activity of rat lung fibroblasts as well as the radiation-induced terminal differentiation of MF progenitor fibroblasts to PMF postmitotic fibrocytes. CONCLUSION: The results indicate the important role of TGF-beta1 in triggering radiation-induced inhibition of clonogenic activity as well as terminal differentiation of rat lung fibroblasts. The data presented support the hypothesis that terminal differentiation is an essential cellular process in the development of radiation-induced fibrosis in the lung. PMID- 10815631 TI - TGF-beta, radiation-induced pulmonary injury and lung cancer. AB - PURPOSE: To determine whether changes in TGF-beta plasma levels during radiation therapy may be useful in predicting radiation-induced pulmonary injury and tumour response in non-small-cell lung cancer (NSCLC) patients. MATERIALS AND METHODS: Plasma TGF-beta was investigated in 27 patients with stage III NSCLC, who were treated with 60 Gy (2Gy/day) radiotherapy with or without carboplatin. TGF-beta was measured prior to beginning radiotherapy and weekly during treatment; evaluated as a ratio between TGF-beta levels obtained during treatment and the pretreatment TGF-beta level. The endpoints of the study were development of symptomatic radiation pneumonitis and tumour response. RESULTS: Nine of the 27 patients developed pneumonitis. The patients who developed pneumonitis had high persistent TGF-beta levels throughout the course of treatment (TGF-beta ratio>1), whereas the TGF-beta levels in patients who did not develop pneumonitis were unchanged or declined towards normal (TGF-beta ratio < 1). Patients who responded to treatment had low or normal TGF-beta levels during treatment compared with patients who failed to respond. Other parameters such as pretreatment TGF-beta values, carboplatin treatment or field size did not appear to have a significant effect, which is probably due to the small number of patients entered in the study. CONCLUSION: This pilot study, with a limited number of patients, suggests the hypothesis that elevated TGF-beta levels during radiotherapy may not only indicate patients with a higher risk of developing pulmonary toxicity but also patients with a higher risk of treatment failure. This remains to be tested in a larger clinical study. PMID- 10815632 TI - The role of T-cells in radiation pneumonitis after bone marrow transplantation. AB - Radiation pneumonitis is a frequent complication of bone marrow transplantation. In limiting the effective dose that can be given, it decreases the chances of cancer cell destruction and of graft acceptance. The incidence of radiation pneumonitis is increased if graft-versus-host disease or infection is present, presumably due to an interaction between T-cell-mediated and radiation-mediated damage. Even in the absence of graft-versus-host disease and infection, we have found that syngeneic T-cells can contribute to radiation pneumonitis in a bone marrow transplant setting. The incidence of radiation pneumonitis was higher after whole-body irradiation and bone marrow transplantation than after whole thorax irradiation. In the former situation, but not the latter, prior thymectomy decreased the incidence of radiation pneumonitis. It is hypothesized that autoreactive T-cells escape induction of self-tolerance during regression of the immune system after whole-body irradiation and, because autoregulatory cells are eliminated, they can contribute to radiation pneumonitis. If this concept is correct, it provides another possible explanation for the sensitivity of the lung to post-transplant pneumonitis and suggests new strategies to limit the incidence of this serious transplant-related complication. PMID- 10815633 TI - Control of radiation-induced pneumopathy and lung fibrosis by angiotensin converting enzyme inhibitors and an angiotensin II type 1 receptor blocker. AB - PURPOSE: This report summarizes our experiences on the protective effect of angiotensin-converting enzyme (ACE) inhibitors, especially captopril and an angiotensin II type 1 receptor blocker on radiation-induced pulmonary injury. METHOD: In the first series of experiments, adult male Sprague Dawley rats were given a single dose of either 20 or 30 Gy of gamma rays to a 35 cm2 right hemithorax port, whilst shielding the left, contralateral, lung. Perfusion scans and autopsies were performed at intervals up to 12 months post-radiation. Three different ACE inhibitors, penicillamine and pentoxifylline were given as radiation protectors and their activity compared. A model of irradiation for total bone marrow transplant (BMT) was used for the second group of experiments. Male WAC/Rij/MCW rats received total-body irradiation and a regimen of cyclophosphamide (CTX) in preparation for bone marrow transplant. The modifiers were two ACE inhibitors, captopril and enalapril, and L-158,809, an angiotensin II (A II) type 1 receptor blocker. All drugs were administered in the rats' drinking water and all were well-tolerated. RESULTS: In the irradiated rats, pulmonary damage progressed from the presence of blebs and detachment from basement membranes of endothelial cells a few days after injury, to severe arteritis and interstitial collagen deposition at 3 months, and then on to severe pneumonitis and extensive pulmonary fibrosis at 6 months. Marked increase of hydroxyproline was also found in the lungs at 6 months. These morphological changes were associated with significant decrease of ACE and plasminogen activator activity (PLA) and a marked increase of prostaglandins (PG12) and thromboxane (Txa2), substances considered as indicators of endothelial pulmonary damage. ACE inhibitors captopril, CL 24817, enalapril and CGS 13945 prevented the markers of endothelial dysfunction. Captopril and CL 24817, which contain a sulphydryl (-SH) radical in their moiety and the AII type 1 receptor blocker, L 158,809, were the most efficient in protecting the lung parenchyma from the inflammatory response and subsequent fibrosis. Penicillamine, an SH-containing compound with weak ACE inhibitory activity was also a strong antifibrotic agent but showed only modest anti-inflammatory properties. Additionally, in the irradiated rats, captopril also reduced the incidence of squamous cell skin carcinomas and subcutaneous sarcomas consequent to the highest doses of radiation. CONCLUSION: ACE inhibitors and one AII type 1 receptor blocker were effective in protecting lungs from radiation-induced pneumonitis and the development of lung fibrosis in two models of rat radiation injury. In the first series of experiments (unilateral irradiation), those ACE inhibitors containing a sulphydryl radical were more effective than those without it. This observation led to the question of whether this protective effect is related to inhibition of AII synthesis or rather to some of the collateral pharmacologic properties of these drugs, such as anti-oxidation or protease inhibition. The AII receptor blocker, however, was shown to be equally effective, if not better, in its antifibrotic capacity than any ACE inhibitor with or without an SH radical, reaffirming the role of AII in modulation of collagen synthesis. PMID- 10815635 TI - DNA damage induced by radiation of different linear energy transfer: initial fragmentation. AB - PURPOSE: To investigate DNA fragmentation as a function of linear energy transfer (LET) after exposure to accelerated ions in the LET range 40-225 keV/microm. MATERIALS AND METHODS: Fragmentation patterns of double-stranded DNA in the range 5 kilobasepairs (kbp) to 5.7 megabasepairs (Mbp) were analysed after irradiation of low-passage GM 5758 normal human fibroblast cells with 60Co-photons, helium ions at 40 keV/microm and high-LET nitrogen ions between 80 and 225 keV/microm. Two separate pulsed-field gel electrophoresis protocols were used, optimized for separation of 1-6 Mbp and 5 kbp to 1.5 Mbp fragments. RESULTS: An increased probability of formation of short and medium-sized DNA fragments was revealed following high-LET irradiation. The DNA double-strand break (dsb) induction yields were, respectively, 5.8 and 6.9-8.8 x 10(-9) dsb bp(-1) Gy(-1) for 60Co photons and ions. The ion yields were some 80-110% higher than those calculated according to a conventional approach, disregarding the fragment distributions. For photons, the yield was 13% higher. The corresponding relative biological effectiveness (RBE) of dsb induction was in the range 1.2-1.5. CONCLUSIONS: A significant non-random contribution to the number of dsb after irradiation with high-LET was confirmed by detailed fragment analysis using pulsed-field gel electrophoresis. The LET had a strong influence on the initial DNA fragment distribution, and hence also on the induction yields measured. However, when the LET was increased to the highest values studied for nitrogen ions, the yield decreased slightly. PMID- 10815634 TI - The effect of dimethyl sulphoxide on the induction and repair of double-strand breaks in human cells after irradiation with gamma-rays and accelerated ions: rapid or slow repair may depend on accessibility of breaks in chromatin of different compactness. AB - PURPOSE: The repair of double-strand breaks (dsb) in mammalian cells is characterized by a rapid phase with a half-life of less than half an hour and a slower phase that lasts for many hours. The proportion of slow repair increase with LET and it has been suggested that the slow repair component consists of more complex damage and is more deleterious to the cells. To see if removal of OH radicals could remove part of the damage in complex dsb and make them easier to repair, human cells were irradiated in the presence of dimethyl sulphoxide (DMSO). METHODS: Induction and repair of dsb were studied by neutral elution in human VH10 cells exposed to y-rays, helium ions (mean LET 40 keV/microm) and 80 and 125 keV/microm monoenergetic nitrogen ions in the presence and absence of 2 M DMSO. RESULTS: Incubation of cells exposed to gamma-rays, 40 keV/microm helium and 80 keV/microm N ions demonstrated that scavenging of OH radicals by DMSO removed most of the rapid repair component. The response to DMSO was less marked after 125 keV/microm nitrogen ions, where about half of the rapid repair was resistant to DMSO. CONCLUSIONS: It is unlikely that the complexity of dsb is responsible for the slow repair because the removal of OH radicals did not make the breaks easier to repair. Instead, it is suggested that rapid and slow repair can be explained on the basis of how different parts of the chromatin are accessible to repair enzymes. PMID- 10815636 TI - Rejoining of DNA fragments produced by radiations of different linear energy transfer. AB - PURPOSE: To analyse the rejoining of DNA double-strand breaks (dsb) produced by high-linear energy transfer (LET) ions, with the specific focus on the influence, on the rejoining estimates, of the way dsb are distributed along chromosomes. MATERIAL AND METHODS: Low passages of normal human fibroblasts (GM5758) were irradiated with 60Co photons, 40 keV/microm helium ions or nitrogen ions with LETs of 80, 125, 175 and 225 keV/microm. The double-stranded DNA fragment distributions, ranging from 5 kbp to 5.7 Mbp, were assayed by pulsed-field gel electrophoresis after repair incubation for 0-22 h. RESULTS: The rejoining was biphasic and the half-times of the two phases were 15 min and 2-3h, respectively, and were independent of LET. Although the majority of breaks were rejoined by the fast phase, both the fraction of dsb rejoined by the slow phase and the fraction of unrejoined dsb at 20-22h increased with increasing LET. CONCLUSIONS: DNA fragment analysis detected LET-dependent differences in the amount of rejoining while the half-times were independent of LET. The majority of dsb were rapidly rejoined even after high-LET irradiation. If fragment-size distribution is not taken into account, both the fraction of breaks rejoined by slow kinetics, and the fraction of unrejoined breaks, can be overestimated when the LET is increased. PMID- 10815637 TI - Effects of deficiency in p53 or bcl-2 on the sensitivity of clonogenic cells in the small intestine to low dose-rate irradiation. AB - PURPOSE: To study the role of p53 and bcl-2 in the response of the small intestine to irradiation delivered at low dose-rate. MATERIALS AND METHODS: Mice homozygous for p53 or bcl-2 deletion (-/-), their respective heterozygotes (+/-), and their wild-type littermates (+/+) including a previously used hybrid strain (B6D2F1), were irradiated to the whole-body using 60Co gamma-rays at 1 Gy h(-1). Crypt survival levels in the small intestine were measured at day 3 after the end of irradiation. RESULTS: Crypt survival levels were higher in p53 -/- mice than in the other p53 genotypes after 25-30 Gy, but not after lower or higher doses. Similar experiments with the three genotypes for bcl-2 status showed lower crypt survival after all doses used in the -/- mice, compared with the +/- and +/+ mice, which were similar in response. The marked degree of curvature in the survival curve observed for the p53 genotypes was also observed in B6D2F1 hybrid mice, was particularly striking in the p53 -/- mice, but was not seen to the same extent in the bcl-2 genotypes. The heterozygotes for p53 or for bcl-2 were nearer in response to their respective +/+ genotypes rather than the -/- genotypes. CONCLUSION: The increased crypt survival levels at some radiation dose levels in the p53 nulls contrasts with the lack of change reported previously using irradiation at high dose-rate. The decreased survival in the bcl-2 nulls is consistent with the known 'survival' function of bcl-2, although bcl-2 expression has not been detected immunohistochemically in this intestinal site. The marked degree of curvature in the dose-response curve at high dose levels for some genotypes was unexpected at this low dose-rate. PMID- 10815638 TI - Tissue lesions caused by microplanar beams of synchrotron-generated X-rays in Drosophila melanogaster. AB - PURPOSE: To examine tissue lesions caused by microplanar beams of synchrotron generated X-rays in Drosophila melanogaster using stereomicroscopy, light and electron microscopy. MATERIALS AND METHODS: Pupae were irradiated by 25-microm wide, 1.175 mm-high parallel microplanes at 100 microm on-centre intervals, at 20, 24, 32, 36, 48 or 72 h of development, with absorbed doses per microplane between 75 and 3,000 Gy. RESULTS: Transverse or longitudinal irradiation with in slice absorbed doses of 75 or 375 Gy caused no recognizable effects. All pupae irradiated at or after 48 h developed normally. Conversely, the development to adulthood was delayed in 90% of pupae irradiated at 24h with doses of 750 Gy. However, neither those pupae nor adults that hatched after pupal irradiation at 48 and 72 h displayed morphological changes. Pupae exposed at 48 h of development to 3,000 Gy developed into adults with sharply delimited lesions in the irradiated microplanes of the compound eye or the cuticle of wings and abdomen. CONCLUSIONS: Post-mitotic eukaryotic cells can survive radiation doses of 3,000 Gy largely undamaged, even at the beginning of the terminal morphogenesis. The extremely sharp delimitation between damaged tissue microplanes and adjacent intact tissues may be relevant for future perspectives of radiosurgery. PMID- 10815639 TI - Pain relief caused by millimeter waves in mice: results of cold water tail flick tests. AB - PURPOSE: To find out if millimeter waves can decrease experimental pain response in mice using cold water tail flick test. MATERIALS AND METHODS: Male Swiss albino mice (15 mice per group) were exposed to continuous millimeter waves at a frequency of 61.22 GHz with incident power densities (IPD) ranging from 0.15 to 5.0 mW/cm2 for 15 min or sham exposed. Latency of tail withdrawal in a cold water (1 +/- 0.5 degrees C) tail flick test was measured before the exposure (baseline) and then four times after the exposure with 15 min breaks. RESULTS: The mean latency of the tail flick response in mice exposed to millimeter waves was more than twice that of sham-exposed controls (p<0.05). This effect was proportional to the power of millimeter waves and completely disappeared at an IPD level of < or = 0.5 mW/cm2. Pretreatment of mice with the opioid antagonist naloxone (1 mg/kg i.p.) blocked the effect of millimeter waves. CONCLUSIONS: Results suggest that the antinociceptive effect of millimeter waves is mediated through endogenous opioids. PMID- 10815640 TI - Technical report. The application of probability-generating functions to linear quadratic radiation survival curves. AB - PURPOSE: To illustrate how probability-generating functions (PGFs) can be employed to derive a simple probabilistic model for clonogenic survival after exposure to ionizing irradiation. METHODS: Both repairable and irreparable radiation damage to DNA were assumed to occur by independent (Poisson) processes, at intensities proportional to the irradiation dose. Also, repairable damage was assumed to be either repaired or further (lethally) injured according to a third (Bernoulli) process, with the probability of lethal conversion being directly proportional to dose. Using the algebra of PGFs, these three processes were combined to yield a composite PGF that described the distribution of lethal DNA lesions in irradiated cells. RESULTS: The composite PGF characterized a Poisson distribution with mean, chiD+betaD2, where D was dose and alpha and beta were radiobiological constants. This distribution yielded the conventional linear quadratic survival equation. To test the composite model, the derived distribution was used to predict the frequencies of multiple chromosomal aberrations in irradiated human lymphocytes. The predictions agreed well with observation. This probabilistic model was consistent with single-hit mechanisms, but it was not consistent with binary misrepair mechanisms. CONCLUSIONS: A stochastic model for radiation survival has been constructed from elementary PGFs that exactly yields the linear-quadratic relationship. This approach can be used to investigate other simple probabilistic survival models. PMID- 10815641 TI - Nocturnal oxygen enrichment in sleep apnoea. AB - We hypothesized that a modest oxygen enrichment, rather than 100% oxygen supplementation as used in previous trials, could result in improvement in ventilatory and cardiac symptoms, in patients with obstructive sleep apnoea (OSA), without jeopardizing the chemostimulant ventilatory drive. This hypothesis was tested in five male patients with OSA in a single-blinded trial consisting of one night spent sleeping in control room air (control night), followed by one night spent sleeping while exposed to air with a 9% enriched oxygen content (oxygen-enriched night). Oxygen enrichment resulted in a significant shift in the oxygen saturation profile towards values of > or = 95% and to decrease desaturation dips throughout the night. The apnoea index decreased from the control night to the oxygen-enriched night from 52.7 +/- 10.4 to 38.9 +/- 9.3; the decrease being greatest for the longest apnoeas (> or = 30 s). Additionally, the cardiovascular status improved. No signs of depressed chemostimulant drive in the oxygen-enriched night were detected. We conclude that nocturnal oxygen enrichment merits consideration for therapeutic trial in the prevention of long apnoeic and desaturation episodes. PMID- 10815642 TI - Left ventricular mass and global function in essential hypertension after antihypertensive therapy. AB - This study compares left ventricular global function in 55 patients (25 with hypertrophy and 30 without hypertrophy) with essential hypertension, whose blood pressure had been stable for longer than 1 year, with that in 35 healthy subjects. Left ventricular global function was calculated using the echocardiographic Doppler index as described by Tei et al. (Tei index). No significant differences were observed in echocardiographic left ventricular systolic function between the three groups. Left ventricular global function differed significantly between the three groups, however, suggesting impaired left ventricular global function even in the absence of left ventricular hypertrophy. Similar changes were observed in the ratio of peak velocity of atrial systole to peak velocity of early diastole during left ventricular inflow waveform (A/E), indicating that this disorder mainly reflects impaired left ventricular diastolic function. Unlike A/E, the Tei index is not affected by increasing age, and, therefore, is appropriate for evaluating left ventricular global function in hypertensive patients, most of whom are middle-aged or older. This index may be useful for determining treatment strategy and evaluating treatment effects. PMID- 10815643 TI - Relationship between changes in volume of the oviductal fluid in the ampulla and the descent of ovulated eggs from the ampulla to the isthmus in mice. AB - Follicular growth and ovulation were induced in mice by administration of pregnant mare serum gonadotrophin (PMSG) followed 2 days later by human chorionic gonadotrophin; the day of PMSG injection was designated as day 0. The volume of the ampulla was measured and the location of the ovulated eggs determined at 06:00, 10:00, 14:00, 18:00 and 22:00 on day 3, and at 02:00 and 06:00 on day 4. The volume of the ampulla and hence oviductal fluid, peaked at 14:00 on day 3 and then declined. In all oviduct samples taken up to 14:00 on day 3, eggs were found exclusively in the ampulla. Thereafter, an increasing number of eggs were observed in the isthmus. Thus, the migration of eggs from the ampulla to the isthmus was concurrent with the decrease in oviductal fluid volume. The peak in the volume of oviductal fluid seen at day 3 is likely to coincide with the opening of the ampullary-isthmic junction of the oviduct. PMID- 10815644 TI - The yield of abnormal unfertilized eggs observed after repeated gonadotrophin induced ovulation. AB - This study investigated whether or not the yield of abnormal eggs produced after repeatedly induced ovulation could be reduced by increasing the interval between gonadotrophin (GTH) administrations. Ovulation was induced, in a mouse model, by subcutaneous injection of pregnant mare serum GTH and human chorionic GTH given once (n = 10), twice (n = 10), three times (n = 10) or four times (n = 10) at intervals of 5 days. In 30 further mice, a fifth induction was performed either 5 (n = 10), 10 (n = 10) or 20 days (n = 10) following the fourth injection. Oviducts were examined microscopically, and the eggs were counted and examined for morphological abnormalities. In the mice who had received four GTH treatments, 33.3% (12 of 36) of the eggs were abnormal. After the fifth induced ovulation performed after an interval of 5, 10 or 20 days, the yield of abnormal eggs was 50.0% (five of 10), 23.1% (nine of 39) and 15.2% (seven of 46), respectively. These results indicate that prolongation of the interval between GTH administrations reduced the yield of abnormal eggs produced following repeatedly induced ovulation. PMID- 10815645 TI - Effect of intrarectal prostaglandin E2 analogue (enprostil) on trinitrobenzenesulphonic acid-induced colitis in rats. AB - Prostaglandins exert a protective effect on colonic mucosa in experimentally induced colitis. This study investigated the effect of enprostil, a prostaglandin E2 (PGE2) analogue, on trinitrobenzenesulphonic acid (TNBS)-induced colitis in rats. Each rat received a rectal enema containing TNBS (30 mg), followed 24 h later by intrarectal once-daily enprostil (200 microg). Enprostil-treated and control rats were killed on day 3 (enprostil group, n = 5; control, n = 6) or day 10 (enprostil group, n = 5; control, n = 5) after TNBS treatment. The area of damaged mucosa of the colon was measured relative to the total colonic area. We also determined the macroscopic score of mucosal damage, and measured PGE2, 6 keto-prostaglandin F1alpha (6-keto-PGF1alpha) and thromboxane B2 (TXB2) concentration in portal vein blood samples. Enprostil significantly reduced both the area of damaged mucosa (including the ulcer area) and the macroscopic score after 3 days' treatment compared with control. Similarly, enprostil significantly reduced plasma concentration of PGE2, 6-keto-PGF1alpha and TXB2 during the acute phase at day 3 of treatment compared with control, but not at day 10. These results suggest that PGE2 enema may have therapeutic potential for treating patients with proctitis or left-sided colitis. PMID- 10815646 TI - Extensive cell death in thymocytes in colon 26-induced cachectic mice. AB - Extensive atrophy has been reported to occur in the thymus in a cancer-burden state but the mechanisms of this atrophy have not been fully elucidated. We investigated changes in the thymus in tumour-bearing mice inoculated with two subclones of the murine colon 26 adenocarcinoma cell line: clone 5 (non cachectic) and clone 20 (cachectic). In clone 20 mice, body weights and thymocyte numbers decreased significantly compared with controls. Flow cytometric analysis of the thymocytes demonstrated that the frequency of single positive cells (CD4+ CD8- and CD4- CD8+) was significantly increased and that of double positive cells (CD4+ CD8+) was significantly decreased in clone 20 mice and, to a lesser extent, in clone 5 mice compared with controls. Serum levels of interleukin 6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were significantly elevated. These results suggested that thymocyte apoptosis was accelerated in the cancer-cachectic state, and increased GM-CSF might be partly responsible for thymic atrophy. PMID- 10815647 TI - An overview of generalized anxiety disorder: disease state--appropriate therapy. AB - OBJECTIVE: This article reviews the prevalence, diagnosis, and treatment of generalized anxiety disorder (GAD). BACKGROUND: Patients with GAD often present to primary care physicians; frequently the disorder manifests with somatic symptoms that have no identifiable physiologic foundation. Accurate diagnosis and treatment often prove elusive, and health care resources are inappropriately consumed in the management of a wide array of complaints, including headache, noncardiac angina, fatigue, insomnia, or abdominal discomfort. Early diagnosis and intervention are critical; GAD is frequently associated with other anxiety and mood disorders, major depressive disorder among them. The differential diagnosis of GAD is complex, including medication side effects and substance related dependence or withdrawal phenomena, as well as endocrine, neurologic, cardiorespiratory, and autoimmune disorders. CONCLUSIONS: GAD is differentiated from adjustment disorder with anxiety because only GAD can manifest without identifiable emotional stressors; it is differentiated from panic disorder largely on the basis of the chronicity of GAD and the episodic, abrupt nature of panic attacks, with the involvement of at least 4 autonomic, cardiopulmonary, neurologic, or other symptoms. In addition to psychotherapy, education, lifestyle modifications, and social support, several pharmacologic agents may be appropriate therapy for GAD. Given the chronic, nonremitting, relapsing character of GAD, use of benzodiazepines, which confer short-term relief, is usually ill advised in long-term treatment because these agents can impair cognitive and psychomotor function, interact with various central nervous system depressants (eg, alcohol), and exhibit substantial potential for abuse, tolerance, dependence, and withdrawal effects. Buspirone and certain antidepressants, including the dual noradrenergic-serotonergic reuptake inhibitor venlafaxine, represent first-line therapy for GAD. PMID- 10815648 TI - Augmentation of standard depression therapy. AB - OBJECTIVE: This article reviews treatment options (eg, augmentation) for depressed patients with suboptimal clinical responses to an antidepressant. BACKGROUND: Approximately one third of patients treated with antidepressants exhibit suboptimal or delayed clinical response to these medications. In such cases, alternative options include switching to another antidepressant or adding a second antidepressant. Augmentation strategies include addition of lithium carbonate, atypical antipsychotics, psychostimulants, thyroid hormone (triiodothyronine), pindolol, or buspirone. CONCLUSIONS: In approximately half of all antidepressant-resistant cases of major depressive disorder, controlled clinical trials have indicated that augmentation with lithium or thyroid hormone is effective. Other reports suggest that central nervous system stimulants may augment antidepressant activity, but their use is constrained by possible abuse potential. Pindolol therapy has been shown to accelerate clinical response in some but not all studies. Finally, the favorable safety and tolerability profile of buspirone, together with its desirable anxiolytic effects, render it a sound therapeutic option in antidepressant augmentation. PMID- 10815649 TI - New directions in the treatment of antidepressant-induced sexual dysfunction. AB - OBJECTIVE: This article reviews options in the treatment of antidepressant induced sexual dysfunction (SD). BACKGROUND: With adoption of structured interviews concerning sexual function, researchers have become increasingly aware that a substantial proportion of patients experience untoward sexual side effects from their antidepressants. As many as half of patients treated with selective serotonin reuptake inhibitors report delayed orgasm (ejaculation), and virtually all patients treated with clomipramine experience anorgasmia. Treatments proven to be effective include yohimbine, sildenafil citrate, buspirone, or other adjuncts, or a temporary drug holiday. SD is most effectively diagnosed and treated in the setting of an empathic physician-patient relationship, which includes frank discussions about sexuality. CONCLUSIONS: Although depressed patients do care about their sexual function, they may be reluctant, for fear of embarrassment, to report SD spontaneously to their physicians. SD is probably underreported and may result in covert noncompliance and attendant relapse into depression. Physicians thus need to assess sexual function during initial evaluation and throughout treatment. The importance of sexual function to sexually active patients with major depression should be weighed carefully when planning antidepressant therapy. A number of viable options exist to prevent or treat SD, including use of relatively new or novel antidepressants and appropriate adjunctive regimens. PMID- 10815650 TI - Preoperative identification of the facial nerve achieved using fast spin-echo MR imaging: can it help the surgeon? PMID- 10815651 TI - Diffusion-weighted MR imaging of multiple sclerosis: added clinical value or "just another pretty face?". PMID- 10815652 TI - What more can MR imaging teach us about brain injury? PMID- 10815653 TI - Spatial relationship between vestibular schwannoma and facial nerve on three dimensional T2-weighted fast spin-echo MR images. AB - BACKGROUND AND PURPOSE: During surgical removal of a vestibular schwannoma, correct identification of the facial nerve is necessary for its preservation and continuing function. We prospectively analyzed the spatial relationship between vestibular schwannomas and the facial nerve using 3D T2-weighted and postcontrast T1-weighted spin-echo (SE) MR imaging. METHODS: Twenty-two patients with a unilateral vestibular schwannoma were examined with MR imaging. The position and spatial relationship of the facial nerve to adjacent tumor within the internal auditory canal (IAC) and cerebellopontine angle cistern (CPA) were assessed on multiplanar reformatted 3D T2-weighted fast spin-echo (FSE) images and on postcontrast transverse and coronal T1-weighted SE images. The entrance of the nerve into the bony canal at the meatal foramen and the nerve root exit zone along the brain stem were used as landmarks to follow the nerve course proximally and distally on all images. RESULTS: The spatial relationship between vestibular schwannoma and facial nerve could not be detected on postcontrast T1-weighted SE images. In 86% of the patients, the position of the nerve in relation to the tumor was discernible on multiplanar reformatted 3D T2-weighted FSE images. In tumors with a maximal diameter up to 10 mm, the entire nerve course was visible; in tumors with a diameter of 11 to 24 mm, only segments of the facial nerve were visible; and in tumors larger than 25 mm, the facial nerve could not be seen, owing to focal nerve thinning and obliteration of landmarks within the IAC and CPA. CONCLUSION: Identification of the facial nerve and its position relative to an adjacent vestibular schwannoma is possible on multiplanar reformatted 3D T2 weighted FSE images but not on postcontrast T1-weighted SE images. Detection of this spatial relationship depends on the tumor's size and location. PMID- 10815654 TI - Neck infection associated with pyriform sinus fistula: imaging findings. AB - BACKGROUND AND PURPOSE: Acute suppurative neck infections associated with branchial fistulas are frequently recurrent. In this study, we describe the imaging findings of acute suppurative infection of the neck caused by a third or fourth branchial fistula (pyriform sinus fistula). METHODS: Imaging findings were reviewed in 17 patients (11 female and six male patients, 2 to 49 years old) with neck infection associated with pyriform sinus fistula. Surgery or laryngoscopic examination confirmed the diagnoses. Fourteen patients had a history of recurrent neck infection and seven had cutaneous openings on the anterior portion of the neck (all lesions were on the left side). Imaging studies included barium esophagography (n = 16), CT (n = 14), MR imaging (n = 2), and sonography (n = 3). RESULTS: A sinus or fistulous tract was identified in eight of 16 patients on barium esophagograms. In 14 patients, CT studies showed the inflammatory infiltration and/or abscess formation along the course of the sinus or fistulous tract from the pyriform fossa to the thyroid gland. In nine patients, CT scans showed the entire course or a part of the sinus or fistulous tract as a tiny spot containing air. MR images showed a sinus or fistulous tract in two patients, whereas sonograms could not depict a sinus or fistulous tract in three patients. All 17 patients were treated with antibiotics. In one patient, the sinus tract was surgically excised, while 15 patients underwent chemocauterization of the sinus or fistulous tract with good outcome. Follow-up was possible for 16 of the 17 patients. CONCLUSION: When an inflammatory infiltration or abscess is present between the pyriform fossa and the thyroid bed in the lower left part of the neck, an infected third or fourth branchial fistula should be strongly suspected. PMID- 10815655 TI - Sonographic findings of the neopharynx after total laryngectomy: comparison with CT. AB - BACKGROUND AND PURPOSE: To our knowledge, sonographic findings in the neopharynx have not been well characterized. We describe our results and assess the role of sonography versus CT in patients who have undergone total laryngectomy. METHODS: We examined 25 patients (24 men and one woman; 44-78 years old) who had had a total laryngectomy. Sonography (with a 10-MHz transducer) and contrast-enhanced CT were performed in all patients. We evaluated the normal shape of the neopharynx and assessed the accuracy of sonography versus CT in detecting tumor recurrence in the neck. RESULTS: The neopharynx appears as a round or ovoid structure on imaging studies. On sonograms, the neopharyngeal wall has five layers of alternating echogenicity: an innermost hyperechoic layer of superficial mucosa, an inner hypoechoic layer of deep mucosa, a middle hyperechoic layer of submucosa, an outer hypoechoic layer of muscle, and an outermost hyperechoic layer of adventitia. On CT scans, the neopharynx appears as a three-layered structure, with an inner hyperdense layer of mucosa, a middle hypodense layer of submucosa, and an outer isodense layer of pharyngeal constrictor muscles. Nine pathologically proved recurrences were found: three local recurrences, one local recurrence with lymph node metastasis, and five cases of lymph node metastasis only. One instance of false-negative lymph node metastasis was seen at sonography and one case of false-positive local recurrence was seen at CT. CONCLUSION: The neopharynx has a unique sonographic appearance, and this imaging technique is useful for detecting local tumor recurrence in the neopharynx in patients who have had a total laryngectomy. PMID- 10815656 TI - Imaging of mucormycosis skull base osteomyelitis. AB - Skull base osteomyelitis (SBO) is typically bacterial in origin and caused by Pseudomonas, although the fungus Aspergillus has also rarely been implicated. SBO generally arises from ear infections and infrequently complicates sinonasal infection. Rhinocerebral Mucor infection is characteristically an acute, fulminant, and deadly infection also affecting the orbits and deep face and is associated with intracranial complications. Bony involvement is uncommon because of the angioinvasive nature of the fungus. More recently, chronic invasive Mucor sinusitis has been described. We report the unusual clinical and imaging features of a patient with biopsy-proven invasive mucormycosis arising from chronic isolated sphenoid sinus disease, who presented with extensive SBO and a paucity of deep facial, orbital, or intracranial involvement. PMID- 10815657 TI - Proton MR spectroscopy in children with bipolar affective disorder: preliminary observations. AB - BACKGROUND AND PURPOSE: Bipolar affective disorder (BPAD) can have its onset during childhood, but the diagnosis may be difficult to establish on the basis of clinical findings alone. Our purpose was to determine whether proton MR spectroscopy can be used to identify abnormalities in the brain of children with BPAD. METHODS: Ten children, ages 6 to 12 years, underwent clinical testing to establish the diagnosis of BPAD. After a drug washout period, all patients underwent MR spectroscopy in which a TE of 135 was used along with a single-voxel placement in both frontal and temporal lobes during a single session. Peaks from N-acetylaspartate (NAA), choline (Cho), glutamate/ glutamine (Glu/Gln), and lipids were normalized with respect to the creatine (Cr) peak to obtain ratios of values of peak areas. These data were compared with those obtained in 10 non-age matched control subjects. To corroborate our data, five children with BPAD also underwent 2D MR spectroscopic studies of the frontal lobes with parameters similar to those used in the single-volume studies. RESULTS: All children with BPAD had elevated levels of Glu/Gln in both frontal lobes and basal ganglia relative to the control group. Children with BPAD had elevated lipid levels in the frontal lobes but not in the temporal lobes. Levels of NAA and Cho were similar for all locations in both groups. Two-dimensional MR spectroscopic studies in five children with BPAD confirmed the presence of elevated lipids in the frontal lobes. CONCLUSION: Our preliminary observations suggest that MR spectroscopy may show abnormalities in children with BPAD not found in unaffected control subjects. It remains to be established whether these abnormalities are a signature of the disease and can be used as a screening test. PMID- 10815658 TI - X-linked adrenoleukodystrophy: the role of contrast-enhanced MR imaging in predicting disease progression. AB - BACKGROUND AND PURPOSE: Early assignment of disease progression among patients with X-linked adrenoleukodystrophy (ALD) is critical for the appropriate selection of effective therapy. We evaluated the association between contrast enhancement on T1-weighted spin-echo MR images and disease progression. METHODS: Clinical charts of patients with X-linked ALD were reviewed for age, availability of MR images of the brain, severity of neurologic impairment, and duration and number of follow-up evaluations. Forty-three male patients with X-linked ALD had undergone multiple MR imaging examinations of the brain that consisted of at least sagittal and axial T1-weighted spin-echo, axial double-echo spin-echo, and contrast-enhanced axial T1-weighted spin-echo imaging. The MR images were reviewed for the presence of contrast enhancement. In addition, global disease burden, as shown by the double-echo spin-echo images, was assessed using a visual scoring method (Loes score). RESULTS: Enhancement was seen on the initial T1 weighted spin-echo MR images of 21 (49%) patients; 18 (86%) of the 21 patients had disease progression revealed by the follow-up evaluations based on MR imaging (Loes) and neurologic scores. No enhancement was seen on the initial T1-weighted spin-echo MR images of 22 (51%) patients; for 18 (82%) of the 22 patients, no evidence of disease progression was revealed by the follow-up evaluations. CONCLUSION: There is a very strong association between the presence of contrast enhancement on T1-weighted MR images and X-linked ALD progression based on clinical evaluation and MR imaging. PMID- 10815659 TI - MR imaging of CNS involvement in children affected by chronic liver disease. AB - BACKGROUND AND PURPOSE: MR imaging sheds new light on CNS involvement in the course of acquired chronic liver disease; however, the exact pathogenetic mechanisms of hepatic encephalopathy and associated MR abnormalities remain unclear. Our purpose was to relate MR signal intensity abnormalities of the CNS to clinical, biochemical, and pathologic features of childhood-onset chronic liver disease. METHODS: Twenty-one patients (12 male and nine female patients) were included in the study; two had Crigler-Najjar disease type 2, 17 had chronic liver disease of different causes, and two had idiopathic copper toxicosis. Twelve patients had histologically proved liver cirrhosis, with a median disease duration of 175 months at the time of MR study. None had clinical symptoms of hepatic encephalopathy. MR imaging was performed using spin-echo T1- and T2 weighted sequences. RESULTS: Eleven patients had abnormal MR imaging findings of the brain revealed by T1-weighted MR sequences; two of the 11 had idiopathic copper toxicosis. The affected sites were the hypothalamus and globus pallidus, presenting symmetrical and bilateral high signal intensities, or the pituitary gland, which appeared homogeneously hyperintense, or both findings. Eight of the 12 patients with cirrhosis had abnormal MR signals of the brain. In these, the median cirrhosis duration was shorter (169 months) than in the remaining four patients with normal MR signals (177 months). A significant correlation was found between abnormal MR signals of the brain and cirrhosis (P = .008) and factor V activity (P = .008). CONCLUSION: MR imaging confirms the presence of abnormal brain signals in the globus pallidus, hypothalamus, and pituitary gland in patients with childhood-onset liver disease in the absence of clinical symptoms of encephalopathy. Signal intensity abnormalities are likely caused by an as yet unidentified metabolic process partially correlated with the severity of liver disease. PMID- 10815660 TI - Early MR features of hypoxic-ischemic brain injury in neonates with periventricular densities on sonograms. AB - BACKGROUND AND PURPOSE: In the early 1980s, diagnosing periventricular leukomalacia (PVL) in neonates by using cranial sonography was possible for the first time. Our purpose was to investigate the possibility of diagnosing PVL in the acute stage by using MR imaging. We evaluated early MR features of hypoxic ischemic brain injury in neonates with periventricular densities (flares) on cranial sonograms to determine the added value of MR imaging over sonography alone for early diagnosis of brain damage. METHODS: In a prospective study, infants who showed flares and/or cysts on sonograms underwent MR imaging during the (sub)acute stage. RESULTS: Fifty infants were classified according to the highest sonographic grade up to the day of MR imaging: 23 infants had sonographic grade 1 (flares < 1 week), 15 had sonographic grade 2 (flares > or = 1 week), four had sonographic grade 3 (small localized cysts), and eight had sonographic grade 4 (extensive periventricular cysts); none had sonographic grade 5 (multicystic leukomalacia) on the day of MR imaging. Overall, the additional information provided by MR imaging (over sonography alone) consisted of the depiction of hemorrhagic lesions in 64% of the infants. Extent and severity of the hemorrhages varied from isolated punctate lesions to extensive hemorrhages throughout the white matter; the latter were followed by cystic degeneration at autopsy in two infants. In nine of the 12 infants with cystic PVL, MR images showed more numerous or more extensive cysts. In addition, in two infants, MR images showed cysts not present on sonograms. In 32% of the infants, MR imaging provided no additional information; in these children, all but one had flares on sonograms whereas MR images showed no abnormalities or a zone of mild periventricular signal change. CONCLUSION: MR imaging can depict the precise site and extent of hypoxic-ischemic brain injury at an earlier stage and allows a wider differentiation of lesions as compared with sonography alone. Hemorrhagic PVL is considered to be rare, but was present in 64% of our study population. PMID- 10815661 TI - Demyelinating plaques in relapsing-remitting and secondary-progressive multiple sclerosis: assessment with diffusion MR imaging. AB - BACKGROUND AND PURPOSE: Conventional MR imaging does not provide specific information that can be reliably associated with the pathologic substrate and clinical status of patients with multiple sclerosis (MS). Our goals were 1) to determine whether the orientationally averaged water diffusion coefficient () can be used to distinguish between plaques of different severity in these patients and 2) to assess possible correlations between values and disease duration, Expanded Disability Status Scale (EDSS) score, and signal intensity on T1-weighted MR images. METHODS: Twenty patients (10 with relapsing-remitting MS and 10 with secondary-progressive MS) and 11 healthy volunteers underwent a combined conventional and diffusion-weighted MR study of the brain. , a parameter that is proportional to the trace of the diffusion tensor, was computed by averaging the apparent diffusion coefficients measured in the x, y, and z directions. measurements were obtained for selected areas of white matter plaques. Differences in among the three groups were tested using analysis of variance. RESULTS: was significantly higher (1.445 +/- 0.129 x 10(-3) mm2/s) in secondary-progressive lesions than in relapsing-remitting lesions (0.951 +/- 0.08), and both values were higher than in normal white matter (0.732 +/- 0.02). There was a significant negative correlation between and the degree of hypointensity on T1-weighted images, and a positive correlation between and both EDSS score and disease duration. CONCLUSION: Our findings suggest that is useful for distinguishing MS lesions of different severities, which are associated with different degrees of clinical disability. PMID- 10815662 TI - Multiple sclerosis: comparison of trace apparent diffusion coefficients with MR enhancement pattern of lesions. AB - BACKGROUND AND PURPOSE: Diffusion-weighted MR imaging and the trace apparent diffusion coefficient (ADC) provide important structural information about tissues. The purpose of this study was to investigate the relationship between trace ADC values and the enhancement pattern of multiple sclerosis (MS) lesions. METHODS: Ninety-six lesions, identified in 24 patients with MS, were characterized by their enhancement pattern on contrast-enhanced T1-weighted MR images. There were 57 nonenhancing lesions (NELs), 28 homogeneously enhancing lesions (HELs), and 11 ring-enhancing lesions (RELs). The trace ADC means for each type of lesion and for normal-appearing white matter (NAWM) were calculated and compared using Student's t-test. RESULTS: The mean trace ADC values for HELs (mean, 7.7 x 1(-10) m2s(-1); SD, 1.4 x 10(-10) m2s(-1)) were less than those for RELs (mean, 1.2 x 10(-9) m2s(-1); SD, 3.5 x 10(-10)m2s(-1)) and NELs (mean, 1.3 x 10(-9) m2(s-1); SD, 2.6 x 10(-10) m2(s-1)). There was a significant difference between the mean trace ADC values of HELs and RELs as well as between those for HELs and NELs. There was also a significant difference in the mean trace ADC values between all lesion types and NAWM (mean, 6.9 x 10(-10) m2s(-1); SD, 5.0 x 10(-11) m2s(-1)). CONCLUSION: We found a predictable relationship between mean trace ADC and the pattern of enhancement in MS lesions, corresponding to reported histopathologic differences in myelination between lesion types and magnetization transfer ratios. PMID- 10815663 TI - Magnetization transfer imaging in the detection of injury associated with mild head trauma. AB - BACKGROUND AND PURPOSE: Most traumatic brain injuries are classified as mild, yet in many instances cognitive deficits result. The purpose of this study was to investigate possible relationships between quantitative magnetization transfer imaging (MTI) and neurocognitive findings in a cohort of patients with mild head trauma but negative findings on conventional MR images. METHODS: We examined 13 patients and 10 healthy volunteers with a standard MR protocol including fast spin-echo and gradient-echo imaging, to which was added quantitative MTI. MTI was performed with a modified gradient-echo sequence incorporating pulsed, off resonance saturation. Both region-of-interest analysis and contour plots were obtained from the MTI data. A subgroup of nine patients was examined with a battery of neuropsychological tests, comprising 25 measures of neurocognitive ability. RESULTS: The magnetization transfer ratio (MTR) in the splenium of the corpus callosum was lower in the patient group as compared with the control group, but no significant reduction in MTR was found in the pons. Individual regional MTR values were significantly reduced in two cases, and contour plot analysis revealed focal areas of abnormality in the splenium of four patients. All the patients showed impairment on at least three measures of the neuropsychological test battery, and in two cases a significant correlation was found between regional MTR values and neuropsychological performance. CONCLUSION: Our results suggest that MTI and contour plot analysis may add sensitivity to the MR imaging examination of patients with traumatic brain injury. PMID- 10815664 TI - Dynamic contrast-enhanced T2-weighted MR imaging of recurrent malignant gliomas treated with thalidomide and carboplatin. AB - BACKGROUND AND PURPOSE: Dynamic, contrast-enhanced MR imaging has allowed quantitative assessment of cerebral blood volume (CBV) in brain tumors. The purpose of our study was to compare postcontrast T1-weighted imaging with dynamic, contrast-enhanced T2*-weighted echo-planar imaging in the evaluation of the response of recurrent malignant gliomas to thalidomide and carboplatin. METHODS: Serial MR imaging was performed in 18 consecutive patients with recurrent malignant gliomas receiving both thalidomide and carboplatin for 12 month periods. Six patients undergoing carboplatin therapy alone were chosen as control subjects. Conventional postcontrast T1-weighted images were compared with relative CBV (rCBV) maps calculated on a pixel-by-pixel basis from dynamic echo planar imaging data. Tumor progression was evaluated clinically using established criteria for malignant gliomas. Studies were performed at 2- to 3-month intervals, and imaging and clinical findings were compared. RESULTS: Tumor response to treatment, based on clinical findings, did not correlate well with conventional imaging findings. The rCBV values decreased significantly in all patients between the start of therapy and the first follow-up in the study group, but not in the control group. The difference in rCBV values between the clinically stable and the progressive group at 12-month follow-up was statistically significant, with the progressive group having higher values. CONCLUSION: Dynamic, contrast-enhanced MR imaging is a valuable adjunct to conventional imaging in assessing tumor activity during antiangiogenic therapy, and correlates better than conventional studies with clinical status and response to therapy. PMID- 10815665 TI - Quantitative measurement of microvascular permeability in human brain tumors achieved using dynamic contrast-enhanced MR imaging: correlation with histologic grade. AB - BACKGROUND AND PURPOSE: Dynamic contrast-enhanced MR imaging may be used to quantify tissue fractional blood volume (fBV) and microvascular permeability. We tested this technique in patients with brain tumors to assess whether these measurements correlate with tumor histologic grade. METHODS: Twenty-two patients with newly diagnosed gliomas underwent MR imaging followed by surgery. Imaging consisted of one pre- and six dynamic postcontrast 3D spoiled gradient-recalled acquisition in the steady state data sets after administration of a single dose (0.1 mmol/kg) of contrast material. Signal intensity changes in blood and tissue were kinetically analyzed using a bidirectional two-compartment model, yielding estimates of fBV (mL/cm3) and microvascular permeability (mL/100 cm3 per minute). Stained tumor specimens were scored on a four-point scale (1 = low grade, 4 = high grade). RESULTS: Histologic examination revealed one grade 1, eight grade 2, seven grade 3, and six grade 4 tumors. fBV values ranged from 0.5% to 13.7%. Permeability values ranged from -0.4 to 18.8, with a strong correlation (r = 0.83) to tumor grade. Despite some overlap between the permeability values of specific tumors from different grades, differences in the mean were statistically significant. There was a weak correlation (r = 0.39) between estimated fBV and tumor grade, and no statistically significant difference among fBV values in any of the groups. CONCLUSION: This relatively simple method of analysis provides quantitative estimates of fBV and microvascular permeability in human brain tumors, with the permeability being predictive of pathologic grade. The technique can be easily implemented on clinical scanners and may prove useful in the assessment of tumor biology and in therapeutic trials. PMID- 10815666 TI - Posttherapeutic intraaxial brain tumor: the value of perfusion-sensitive contrast enhanced MR imaging for differentiating tumor recurrence from nonneoplastic contrast-enhancing tissue. AB - BACKGROUND AND PURPOSE: Differentiation of tumor recurrence from treatment related changes may be difficult with conventional MR imaging when newly enhancing lesions appear. Our aim was to determine the value of perfusion sensitive contrast-enhanced MR imaging for differentiating recurrent neoplasm from nonneoplastic contrast-enhancing tissue. METHODS: Twenty patients in whom new enhancing lesions developed within irradiated regions were examined prospectively with perfusion-sensitive contrast-enhanced MR imaging. Twelve of them also underwent thallous chloride Tl 201 single-photon emission tomography (201Tl-SPECT). Normalized relative cerebral blood volume (rCBV) ratios and thallium indexes were evaluated to determine whether the new enhancing lesions were recurrent or not. Five instances of tumor recurrence and one of radiation necrosis were verified histologically; in the others, tumor recurrence was distinguished by lesions that progressively increased in size on serial MR examinations over at least 5 months, and nonneoplastic contrast-enhancing tissue was distinguished by lesions that disappeared or decreased in size on serial MR studies over at least 9 months. RESULTS: When normalized rCBV ratios were higher than 2.6 or lower than 0.6, enhancing lesions were either recurrent (n = 5) or nonneoplastic contrast-enhancing tissue (n = 3), respectively. All nonneoplastic contrast-enhancing tissue had a low thallium index, whereas three of four recurrent lesions had a high index. CONCLUSION: An enhancing lesion with a normalized rCBV ratio higher than 2.6 or lower than 0.6 may suggest tumor recurrence or nonneoplastic contrast-enhancing tissue, respectively. In these cases, further examination with 201Tl-SPECT may not be necessary. However, when the normalized rCBV ratio is between 0.6 and 2.6, 201Tl-SPECT may be useful in making the differentiation. PMID- 10815667 TI - Reproducibility of visual activation in functional MR imaging and effects of postprocessing. AB - BACKGROUND AND PURPOSE: Functional MR imaging studies of the brain should be interpreted in the context of their reproducibility. We assessed the reproducibility of visual activation measured by functional MR imaging and analyzed the effect of image transformation to standard space. METHODS: Seven healthy volunteers were studied twice with echo-planner functional MR imaging at 1.5 T during visual stimulation. The studies were separated by an interval of 2 to 7 days. Functional images were analyzed after spatial normalization to the space described by Talairach and Tournoux and/or after coregistration of the images of the second study with the images of the first study. The number of active voxels for each study was determined at three thresholds. In addition, the change in the center of the mass of activation, the mean change in signal intensity, and the mean t value within the activated area were measured. These reproducibility indexes were calculated for the spatially normalized and nonnormalized data for each subject. RESULTS: Variations in visual activation were observed between the two studies in the same individual as well as across subjects. There was no evidence of an effect from image transformation on reproducibility on any of the measures. CONCLUSION: Our findings show that the reproducibility of activation in functional MR imaging may be much more variable across subjects than suggested in previous studies. The use of different types of image transformation (coregistration, spatial normalization) does not significantly affect the reproducibility of visual activation. PMID- 10815668 TI - Effects of a phonologically driven treatment for dyslexia on lactate levels measured by proton MR spectroscopic imaging. AB - BACKGROUND AND PURPOSE: Dyslexia is a language disorder in which reading ability is compromised because of poor phonologic skills. The purpose of this study was to measure the effect of a phonologically driven treatment for dyslexia on brain lactate response to language stimulation as measured by proton MR spectroscopic imaging. METHODS: Brain lactate metabolism was measured at two different time points (1 year apart) during four different cognitive tasks (three language tasks and one nonlanguage task) in dyslexic participants (n = 8) and in control participants (n = 7) by using a fast MR spectroscopic imaging technique called proton echo-planar spectroscopic imaging (1 cm3 voxel resolution). The age range for both dyslexic and control participants was 10 to 13 years. Between the first and second imaging sessions, the dyslexic boys participated in an instructional intervention, which was a reading/science workshop. RESULTS: Before treatment, the dyslexic boys showed significantly greater lactate elevation compared with a control group in the left anterior quadrant (analysis of variance, P = .05) of the brain during a phonologic task. After treatment, however, brain lactate elevation was not significantly different from that of the control group in the left anterior quadrant during the same phonologic task. Behaviorally, the dyslexic participants improved in the phonologic aspects of reading. CONCLUSION: Instructional intervention that improved phonologic performance in dyslexic boys was associated with changes in brain lactate levels as measured by proton echo planar spectroscopic imaging. PMID- 10815669 TI - Comparison of fluid-attenuated inversion-recovery MR imaging with CT in a simulated model of acute subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Because MR imaging is becoming integral to the evaluation and treatment of very early stroke, it is critical to prove that MR imaging is at least as sensitive to acute subarachnoid hemorrhage (SAH) as is CT. The present study was conducted to evaluate the possibility of detecting a small amount of acute SAH diluted by CSF not revealed by CT but identified on fluid-attenuated inversion-recovery (FLAIR) MR images in an in vitro study. METHODS: Acute SAH was simulated with mixtures of artificial CSF and arterial blood (hematocrit [Hct], 45%) ranging from 0% to 100% by volume. We scanned these phantoms with CT and turbo-FLAIR MR imaging (9000/119 [TR/effective TE]; inversion time, 2200 ms; echo train length, 7), and we measured T1 and T2 relaxation times of these phantoms at temperatures within 36 degrees C to 37 degrees C. Plots of CT value from the different blood/water mixture ratios versus Hct were generated and correlated with the average CT value from normal cortex. We measured T1 and T2 relaxation times of these phantoms and normal cortex and generated T2 relaxation curves as a function of effective TE for a specific inversion time (2200), and determined the TR (9000) for the turbo-FLAIR sequence by using a theoretical equation for the turbo inversion recovery signal intensity. RESULTS: Above a Hct of 27% blood, the mixture was denser on CT scans than was the normal cortex. At a selected time longer than an effective TE of 120, above a Hct of 22.4% blood, the mixture was more hyperintense than the normal cortex on turbo-FLAIR images. At selected times longer than an effective TE of 160, above a Hct of 9% blood, the mixture was more hyperintense than was the normal cortex. CONCLUSION: FLAIR imaging is more sensitive than CT in the detection of a small amount of acute SAH diluted by CSF at selected appropriate TE, as determined in an in vitro study. PMID- 10815670 TI - A high-resolution fast spin-echo inversion-recovery sequence for preoperative localization of the internal globus pallidus. AB - A fast spin-echo inversion-recovery (FSE-IR) sequence is described for its utility regarding surgical planning for patients with Parkinson's disease (PD) who are undergoing microelectrode-guided internal globus pallidus (GPi) ablation. Images from thirty-seven adult patients with PD were reviewed and visualization of the GPi, globus pallidus externa (GPe), and the intervening lamina was noted. High-resolution images were acquired from all patients despite the external hardware and the patients' movement disorder. In all cases, the conventional surgical trajectory, determined indirectly by a fixed measurement from the anteroposterior commissure line, was modified by the ability to visualize the GPi and optic tract directly. This sequence facilitated accurate stereotactic targeting. PMID- 10815671 TI - Variations of the superficial middle cerebral vein: classification using three dimensional CT angiography. AB - BACKGROUND AND PURPOSE: Classification of variations of the superficial middle cerebral vein (SMCV) remains ambiguous. We propose a new classification system based on embryologic development for preoperative examination. METHODS: Three dimensional CT angiography was used to evaluate 500 SMCVs (in 250 patients). The outflow vessels from the SMCV were classified into seven types on the basis of embryologic development. The 3D CT angiograms in axial stereoscopic and oblique views and multiple intensity projection images were evaluated by the same neurosurgeon on two occasions. Inconsistent interpretations were regarded as equivocal. RESULTS: Three-dimensional CT angiography clearly depicted the SMCV running along the lesser wing or the middle cranial fossa. However, the outflow vessel could not be confirmed as the sphenoparietal, cavernous, or emissary type in 39 (8%) of the sides. SMCVs running in the middle cranial fossa to join the transverse sinus or superior petrosal sinus were accurately identified. SMCVs were present in 456 sides: 62% entered the sphenoparietal sinus or the cavernous sinus and 12% joined the emissary vein. Nine vessels were the superior petrosal type, 10 the basal type, 12 the squamosal type, and 44 the undeveloped type. CONCLUSION: Three-dimensional CT angiography can depict the vessels and their anatomic relationship to the bone structure, allowing identification of the SMCV variant in individual patients. Preoperative planning for skull base surgery requires such information to reduce the invasiveness of the procedure. With the use of our classification system, 3D CT angiography can provide exact and practical information concerning the SMCV. PMID- 10815672 TI - An unusual variant of the fronto-orbital artery. AB - A patient with a rare variation of the fronto-orbital artery is presented. In this case, the fronto-orbital artery arises from the A1 segment of the contralateral anterior cerebral artery. The angiographic findings in this case are illustrated and discussed. The importance of recognizing this variant is related to the planning of surgery or endovascular therapy in the anterior cerebral artery region. PMID- 10815673 TI - Bilateral thoracic bifurcation of the common carotid artery associated with Klippel-Feil anomaly. AB - We report the case of a 72-year-old man with bilateral intrathoracic carotid bifurcations associated with a Klippel-Feil anomaly. The left and right carotid bifurcations were located at levels corresponding to the second and fourth thoracic vertebrae, respectively. A possible association between low carotid bifurcation and the Klippel-Feil anomaly is suggested. PMID- 10815674 TI - Orbital Doppler sonography findings in cases of brain death. AB - In this study, we investigated the blood flow velocity changes in orbital arteries by using Doppler sonography in eight patients with brain death. Peak systolic and end-diastolic velocities and resistive indices of the ophthalmic and central retinal arteries were evaluated. We observed the absence or reversal of end-diastolic blood flow in these arteries. To our knowledge, this finding has not been previously reported to be associated with brain death. PMID- 10815675 TI - Diffusion-weighted MR imaging offers no advantage over routine noncontrast MR imaging in the detection of vertebral metastases. AB - BACKGROUND AND PURPOSE: Diffusion-weighted MR imaging of the spine has been used to differentiate benign from pathologic vertebral body compression fractures. We sought to determine the utility of diffusion-weighted MR imaging in the detection of vertebral metastases and to compare it with conventional noncontrast T1- and T2-weighted MR imaging. METHODS: Fifteen patients with metastases to the spine were studied using conventional MR imaging and diffusion-weighted imaging. Blinded review of all images was undertaken, and patients were categorized according to whether they had focal or multiple lesions. The signal intensity of the lesions was compared on T1-, T2- (fast spin-echo), and diffusion-weighted images. RESULTS: In five patients with focal disease, metastases were hypointense on T1-weighted images; hypointense (n = 2), isointense (n = 1), or hyperintense (n = 2) on T2-weighted images; and hypointense (n = 3) or hyperintense (n = 2) on diffusion-weighted images with respect to presumed normal bone marrow. In 10 patients with disease in multiple sites, all lesions were hypointense on T1 weighted images; hypointense (n = 2), isointense (n = 4), hyperintense (n = 2), or mixed (n = 2) on T2-weighted images; and hypointense (n = 5), hyperintense (n = 3), or mixed (n = 2) on diffusion-weighted images with respect to presumed normal bone marrow. CONCLUSION: As used in this study, diffusion-weighted MR imaging of the spine showed no advantage in the detection and characterization of vertebral metastases as compared with noncontrast T1-weighted imaging, but was considered superior to T2-weighted imaging. PMID- 10815676 TI - Spinal intradural capillary hemangioma: MR findings. AB - We report a case of a spinal intradural capillary hemangioma. On MR images, a well-circumscribed intradural mass was detected at the T8-T9 level. The signal intensity of the mass relative to the spinal cord was isointense on T1-weighted images, hyperintense on T2-weighted images, and showed homogeneous, strong enhancement on contrast-enhanced T1-weighted images. The mass had both extramedullary and intramedullary components. PMID- 10815677 TI - Cerebral aneurysms treated by Guglielmi detachable coils: evaluation with diffusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: The most frequent and serious complications of endovascular treatment of intracranial aneurysms with Guglielmi detachable coils (GDCs) are ischemic lesions caused by thromboembolic events. Diffusion-weighted MR imaging appears to be the most sensitive technique for detecting early ischemic phenomena. We evaluated this technique for the detection of brain changes in patients who underwent GDC treatment of aneurysms. METHODS: Twenty patients with a cerebral aneurysm were studied with diffusion-weighted imaging before and after endovascular treatment with GDCs. Aneurysms were located in the anterior (n = 16) or posterior (n = 4) circulation. Bleeding had occurred in 11 patients. MR studies, including fast fluid-attenuated inversion recovery (FLAIR) and diffusion-weighted sequences, were scheduled before, 2 to 4 hours after, and 48 hours after treatment. MR images, including apparent diffusion coefficient (ADC) maps, were assessed for the presence of acute ischemic stroke lesions. RESULTS: In all patients, the aneurysm was excluded without neurologic worsening. In 18 patients, diffusion-weighted and FLAIR images showed no evidence of recent ischemic lesions after treatment. In one patient, an asymptomatic frontobasal hyperintense signal on diffusion-weighted images with a drop of ADC values corresponding to an acute ischemic lesion was observed. In another patient, multiple silent lesions were seen on diffusion-weighted images after embolization. These silent lesions were not all located in the vascular territory of the aneurysm's parent artery. CONCLUSION: This preliminary study suggests that diffusion-weighted MR imaging is a potentially useful tool for monitoring patients after endovascular treatment of a cerebral aneurysm. While small asymptomatic lesions can be observed on these images after embolization, their exact prevalence should be evaluated in a larger series. PMID- 10815678 TI - Gadopentetate dimeglumine as a contrast agent in common carotid arteriography. AB - BACKGROUND AND PURPOSE: Despite improvements in noninvasive imaging, some patients with contraindications to iodine-based contrast material still require angiography for the evaluation of carotid stenosis. Our aim was to assess the utility of gadopentetate dimeglumine as an intraarterial contrast agent in common carotid angiography. METHODS: Twelve patients with suspected carotid artery stenosis were enrolled in the study. In addition to the standard injection sequences with iohexol, common carotid arteriograms were obtained after administration of gadopentetate dimeglumine. Neurologic status and vital signs were monitored during and for 6 hours after the examination. For each injection, five independent observers, blinded to the contrast agent used, measured the percentage of carotid stenosis and assessed their confidence in grading the stenosis, the overall quality of the examination, and, in cases of decreased quality, the reason(s) for it. Statistical analysis was done with paired and unpaired t-tests with equal variances. RESULTS: No patient had an adverse clinical outcome, and measurements of carotid artery stenosis showed no statistically significant differences between the gadopentetate dimeglumine and iohexol examinations. Overall image quality and observer confidence in measurements of stenosis on the gadolinium-based studies were slightly but significantly lower than those of identical iodine-based studies. CONCLUSION: Gadopentetate dimeglumine may be an alternative to iodine in selected patients undergoing carotid angiography. Although overall image quality of the gadolinium studies is slightly inferior to that of the iohexol studies, measurements of carotid artery stenosis are similar for the two examinations. PMID- 10815679 TI - Iatrogenic arterial spasm relieved by intraarterial mannitol infusion. AB - Catheter placement for blood brain-barrier disruption and enhanced chemotherapy delivery can sometimes trigger arterial spasm of moderate-to-severe degree. A slow infusion of a small quantity of intraarterially administered mannitol (10 mL of 25% mannitol) was evaluated as a means to obtain a rapid resolution of catheter placement-induced spasm. We prospectively report 12 consecutive cases of blood brain-barrier disruption among patients who developed catheter placement induced spasm that was treated by this means without side effects, resulting in rapid resolution of spasm. PMID- 10815680 TI - Elective stenting of symptomatic middle cerebral artery stenosis. AB - Percutaneous balloon angioplasty has been found to be useful for the treatment of intracranial atherosclerotic arterial stenosis. Nonetheless, an ongoing risk of this procedure is arterial dissection, which increases the hazards of acute closure, stroke, and restenosis. Stenting of the intracranial vasculature recently has been shown to be feasible in a variety of circumstances. To our knowledge, however, stenting of the middle cerebral artery has not been possible until now primarily because of difficulty with tracking stents across the carotid siphon. We describe the successful treatment of a symptomatic middle cerebral artery stenosis achieved using a balloon-expandable flexible coronary stent. PMID- 10815681 TI - Remote vascular catastrophes after neurovascular interventional therapy for type 4 Ehlers-Danlos Syndrome. AB - Type 4 Ehlers-Danlos Syndrome (EDS 4) is the most malignant form of Ehlers-Danlos Syndrome, often accompanied by neurovasacular complications secondary to vessel dissection or aneurysms. The fragile nature of connective tissue in these patients makes exovascular and endovascular treatment hazardous. We have treated four patients with EDS 4 over the last 8 years by using neuroendovascular procedures. Two of these individuals suffered remote vascular injuries around the time of their procedures and ultimately died. The circumstances surrounding their deaths will make up the body of this report. PMID- 10815682 TI - In re: Barkovich AJ. MR and CT evaluation of profound neonatal and infantile asphyxia. AJNR Am J Neuroradiol 1992; 13:959-72. PMID- 10815683 TI - High-field, high-resolution MR imaging of the human idusium griseum. PMID- 10815684 TI - Detachable balloon embolization: safety balloon technique. PMID- 10815685 TI - In Re: The Presyrinx state: a reversible myelopathic condition that may precede syringomyelia. PMID- 10815686 TI - Curved planar reformatted CT angiography: utility for the evaluation of aneurysms at the carotid siphon. PMID- 10815687 TI - Experimental arteriovenous malformations modeling in laboratory sheep versus swine. PMID- 10815688 TI - In Re: Tokumaru AM, Barkovich AJ, O'uichi T, Matsuo T, Kutano S. The evolution of cerebral blood flow in the developing brain: evaluation with MR imaging. AJNR Am J Neuroradiol 1999; 20:845-52. PMID- 10815689 TI - Glutathione S-transferase theta 1 gene deletion and risk of acute myeloid leukemia. AB - Individuals with a homozygous deletion of the glutathione S-transferase theta 1 (GSTT1) gene lack GSTT1 enzymatic detoxification of environmental carcinogens by conjugation with glutathione. The GSTT1 gene deletion has been associated with carcinogen-induced chromosomal changes in lymphocytes, and some but not all epidemiological evidence has suggested that the GSTT1 gene deletion may increase susceptibility to myelodysplasia. We conducted a case-control study to test whether individuals with an inherited homozygous deletion of the GSTT1 gene are at increased risk of acute myeloid leukemia (AML). The GSTT1 and GST mu 1 (GSTM1) genotypes were determined by PCR using lymphocyte or bone marrow DNA from 297 AML patients and 152 controls. AML patients were selected from Southwest Oncology Group clinical studies, and controls were identified by random digit dialing in Washington state. No association was observed between the GSTT1 gene deletion and AML [race-adjusted odds ratio (OR), 0.94; 95% confidence interval (CI), 0.55 1.60] or between the GSTM1 gene deletion and AML (race-adjusted OR, 1.26; 95% CI, 0.85-1.88). Patients with secondary AML had a slightly higher prevalence of the GSTT1 and GSTM1 gene deletions compared with de novo AML patients or controls, but this was consistent with chance. Exploratory analyses of AML cytogenetics suggested a few associations, i.e., between the GSTT1 gene deletion and trisomy 8, and between the GSTM1 gene deletion and non-8 trisomies or inv(16). These results do not support the hypothesis that the GSTT1 gene deletion is related to the incidence of AML. PMID- 10815690 TI - Cigarette smoking, N-acetyltransferase 2 acetylation status, and bladder cancer risk: a case-series meta-analysis of a gene-environment interaction. AB - Tobacco use is an established cause of bladder cancer. The ability to detoxify aromatic amines, which are present in tobacco and are potent bladder carcinogens, is compromised in persons with the N-acetyltransferase 2 slow acetylation polymorphism. The relationship of cigarette smoking with bladder cancer risk therefore has been hypothesized to be stronger among slow acetylators. The few studies to formally explore such a possibility have produced inconsistent results, however. To assess this potential gene-environment interaction in as many bladder cancer studies as possible and to summarize results, we conducted a meta-analysis using data from 16 bladder cancer studies conducted in the general population (n = 1999 cases), Most had been conducted in European countries. Because control subjects were unavailable for a number of these studies, we used a case-series design, which can be used to assess multiplicative gene-environment interaction without inclusion of control subjects. A case-series interaction odds ratio (OR) > 1.0 indicates that the relationship of cigarette smoking and bladder cancer risk is stronger among slow acetylators as compared with rapid acetylators. We observed an interaction between smoking and N-acetyltransferase 2 slow acetylation (OR, 1.3; 95% confidence interval, 1.0-1.6) that was somewhat stronger when analyses were restricted to studies conducted in Europe (OR, 1.5; confidence interval, 1.1-1.9), a pooling that included nearly 80% of the collected data. Using the predominantly male European study population and assuming a 2.5-fold elevation in bladder cancer risk from smoking, we estimated that the population attributable risk percent was 35% for slow acetylators who had ever smoked and 13% for rapid acetylators who had ever smoked. These results suggest that the relationship of smoking and bladder cancer is stronger among slow acetylators than among rapid acetylators. PMID- 10815691 TI - A cumulative case-control study of risk factor profiles for oncogenic and nononcogenic cervical human papillomavirus infections. AB - Human papillomaviruses (HPVs) play an essential role in the etiology of cervical cancer, but besides an established role for sexual transmission, little is known about other risk factors for HPV infection. Risk factors for nononcogenic, oncogenic, and HPV 16 cervical infections were investigated using a cumulative case-control approach nested in an ongoing cohort study of low income women from Sao Paulo, Brazil. HPV DNA was detected and typed by the MY09/11 PCR protocol. Risk factor information was obtained via interviews. In a case-control analysis, we compared women who harbored infections with exclusively nononcogenic types (n = 123), exclusively oncogenic types (n = 94), and any HPV 16 (n = 60) to women remaining HPV-negative (n = 512) throughout 1 year of follow-up. A strong negative association was found between age and oncogenic infections, but not with nononcogenic infections. Oral contraceptive use was strongly and exclusively associated with oncogenic and HPV 16 infections. Markers of sexual activity were associated with all types of infections, although with varying strengths. Our results suggest some important differences in the epidemiological correlates of HPV infection according to oncogenicity that may have implications for the planning of specific preventive strategies aiming at reduction of cervical cancer risk. PMID- 10815692 TI - Intakes of fruits, vegetables, and related nutrients and the risk of non Hodgkin's lymphoma among women. AB - Non-Hodgkin's lymphoma is etiologically related to suppressed immune status, and certain nutrients found in fruits and vegetables have been associated with increased immune responses. However, limited information exists on associations between intake of fruits, vegetables, and related nutrients and non-Hodgkin's lymphoma risk. We thus examined these associations among 88,410 women in the Nurses' Health Study cohort who were aged 34-60 years in 1980 and provided dietary information in 1980. During 14 years of follow-up, we documented 199 incident cases of non-Hodgkin's lymphoma. Higher intake of fruits and vegetables was associated with a lower risk of non-Hodgkin's lymphoma (P for trend = 0.02); the multivariate relative risk (RR) was 0.62 [95% confidence interval (CI), 0.38 1.02] for women who consumed greater than or equal to six servings per day as compared with those consuming less than three servings per day. When fruits and vegetables were examined separately, intake of vegetables rather than fruits was most clearly associated with a reduced risk (P for trend = 0.02 for vegetables; P for trend = 0.16 for fruits); compared with those consuming less than one serving per day, the multivariate RRs were 0.62 (95% CI, 0.35-1.07) for women who consumed greater than or equal to three servings per day of vegetables and 0.70 (95% CI, 0.44-1.12) for women who consumed this amount of fruits. Higher intake of cruciferous vegetables was also associated with a decreased risk (P for trend = 0.03); the multivariate RR was 0.67 (95% CI, 0.40-1.11) for women who consumed greater than or equal to five servings per week as compared with those consuming less than two servings per week. These associations were slightly attenuated when we additionally adjusted for intake of beef, pork, or lamb as a main dish. Intake of dietary fiber from vegetable sources was related to a reduced risk; the multivariate RR was 0.54 (95% CI, 0.34-0.87) for women in the highest quintile as compared with those in the lowest quintile (P for trend = 0.01), and it was slightly attenuated with additional adjustment for saturated and trans unsaturated fats. However, we observed no associations between intakes of specific dietary carotenoids, vitamins A, C, E, and folate, and non-Hodgkin's lymphoma risk. Higher intake of vegetables, particularly cruciferous vegetables, may reduce the risk of non-Hodgkin's lymphoma among women. PMID- 10815693 TI - Organochlorines and endometrial cancer risk. AB - There is concern that persistent environmental pollutants such as dichlorodiphenyltrichloroethane (DDT) and polychlorinated biphenyls (PCBs) increase breast cancer risk, at least partially through estrogenic effects. Because the endometrium is more sensitive to estrogenic stimulation than the breast, such a carcinogenic effect should be more pronounced in the endometrium than the breast. In a population-based case-control study in Sweden, we measured serum concentrations of 10 chlorinated pesticides and 10 PCB congeners in 154 endometrial cancer cases and 205 population controls. Information on potential confounders was obtained by mailed questionnaires. We used logistic regression to calculate odds ratios (ORs) as measures of relative risk. We performed analyses for lipid-adjusted concentrations of each individual substance and after grouping substances according to putative hormonal effects. We found no significant associations of increasing levels of pesticide or PCB exposure with endometrial cancer risk. The multivariate OR was 1.0 (95% confidence interval, 0.6-2.0; P for trend, 0.78) for the highest compared with the lowest quartile of dichlorodiphenyldichloroethylene (DDE), the predominant dichlorodiphenyltrichloroethane metabolite. Corresponding ORs were 1.0 for hexachlorobenzene, 0.9 for beta-hexachlorocyclohexane, 1.4 for oxychlordane, and 1.2 for trans-nonachlor. Analyses of substances grouped by putative hormonal effect also showed no associations with endometrial cancer risk. For all estrogenic compounds, the OR for the highest compared with the lowest quartile was 1.1 (95% confidence interval, 0.6-2.2; P for trend, 0.90). Our data do not support the hypothesis that the organochlorine exposure studied increases the risk for endometrial cancer. PMID- 10815694 TI - Quantitative grading of rat esophageal carcinogenesis using computer-assisted image tile analysis. AB - Our objective was to grade, by computer-assisted quantitative image tile analysis, the intraepithelial neoplasia (also called dysplasia) that develops in esophagi of rats given N-nitrosomethybenzylamine (NMBA) for 5 weeks. To perform image tile analysis, the computer divides the video image of the neoplastic epithelium into a row of contiguous small rectangular images, or "tiles," 84 x 292 microm in size, and quantitatively measures four selected tissue features within each image tile. The computer then calculates a tile grade for each image tile as the weighted sum of the four feature measurements, transformed into statistical Z-scores, the weights being determined by Fisher linear discriminant analysis of 300 tile grades of the neoplastic epithelium referenced to the mean tile grade (MTG) of 300 image tiles of normal epithelium. The two grading parameters, MTG and the percentage of tile grades exceeding the MTG of normal epithelium by >4 SD units (%TG>4SD), were validated as endpoints for screening chemopreventive agents in the rat NMBA-induced esophageal carcinogenesis model in two ways: (a) after NMBA treatment, %TG>4SD developed in parallel with tumor incidence and tumor multiplicity (number of papillomas/tumor-bearing rat); and (b) placing the chemopreventive phenethylisothiocyanate in the food of NMBA treated rats produced parallel reductions in MTG, tumor incidence, and tumor multiplicity. Both MTG and %TG>4SD, measured by quantitative image tile analysis, are sensitive and objective continuous parametric response variables expressed to three significant figures, with wide dynamic range, that may be evaluated by t tests to compare tissue neoplastic changes before and after treatment with a chemopreventive agent. PMID- 10815695 TI - Collection of buccal cell DNA using treated cards. AB - We devised a simple, noninvasive, cost-efficient technique for collecting buccal cell DNA for molecular epidemiology studies. Subjects (n = 52) brushed their oral mucosa and expectorated the fluid in their mouths, which was applied to "Guthrie" cards pretreated to retard bacterial growth and inhibit nuclease activity (IsoCode, Schleicher and Schuell, Keene, NH). The cards are well-suited for transport and storage because they dry quickly, need no processing, and are compact and lightweight. We stored the samples at room temperature for 5 days to mimic a field situation and then divided them into portions from which DNA was extracted either immediately or after storage for 9 months at room temperature, 20 degrees C, or -70 degrees C. The fresh samples had a median yield of 2.3 microg of human DNA (range, 0.2-53.8 microg), which was adequate for at least 550 PCR reactions. More than 90% of the samples were amplified in all three beta globin gene fragment assays attempted. DNA extract frozen for 1 week at -20 degrees C also performed well. Stored samples had reduced DNA yields, which achieved statistical significance for room temperature and -70 degrees C, but not -20 degrees C, storage. However, because all of the stored samples tested were successfully amplified, the observed reduction may represent tighter DNA fixation to the card over time rather than loss of genetic material. We conclude that treated cards are an alternative to brushes/swabs and mouth rinses for the collection of buccal cell DNA and offer some advantages over these methods, particularly for large-scale or large-scale or long-term studies involving stored samples and studies in which samples are collected off-site and transported. Future studies that enable direct comparisons of the various buccal cell collection methods are needed. PMID- 10815696 TI - BRCA1 susceptibility markers and postmenopausal breast cancer: the Iowa Women's Health Study. AB - Much research on early-onset breast cancer families has been performed and has shown that breast cancer in many of these families is linked to either BRCA1 or BRCA2. Fewer studies have examined the role of genetic predisposition in postmenopausal breast cancer. A nested case-control family study of breast cancer was conducted within the Iowa Women's Health Study, a population-based prospective study of 41,836 postmenopausal women. Probands were 251 incident cases diagnosed between 1988 and 1989. Three-generation pedigrees were developed through mailed questionnaires. From this collection of pedigrees, thirteen were identified for more detailed genetic analysis. Sibling-pair linkage analyses were performed using polymorphic markers in candidate regions in these 13 families with multiple cases of breast and other cancers. Four of the DNA markers are located on chromosome 17, and two of these (D17S579 and THRA1) flank the BRCA1 locus. Significant evidence for linkage to D17S579 was obtained in the total sample, in a model without inclusion of covariates or age at onset (P = 0.005), and in a model adjusted for five measured covariates and for variable age at onset (P = 0.008). Complete sequencing of the BRCA1 gene in these families, including all intron/exon boundaries, failed to reveal any mutations in 24 women with breast cancer from the 13 families. These data suggest that in some families identified by postmenopausal breast cancer cases, breast cancer risk may be mediated by a gene (or genes) in the BRCA1 region, but not BRCA1 itself. PMID- 10815697 TI - Breast and cervical cancer screening: associations with personal, spouse's, and combined smoking status. AB - The objective of this study was to examine the association of women's cancer screenings with both personal and spouses' smoking status, as well as with the broader context of household smoking, in a United States national-level sample of women aged 42-75 years. Data were from the 1994 National Health Interview Survey Health Promotion Supplement. The sample included 1586 women who reported they were married and living with a spouse in a two-person household. Three measures of smoking status were used: personal smoking status, smoking status of spouse, and household smoking status (self and spouse smoked, spouse only smoked, self only smoked, and both nonsmokers). Using logistic regression modeling, associations were examined between the smoking status measures and three cancer screening indicators: mammogram < or =2 years, clinical breast exam < or =2 years, and Pap test < or =3 years. The both nonsmokers group consistently had the highest screening rates for all three exams. The spouse only smoking group was 10 12% less likely to obtain all three cancer screening tests compared to the both nonsmokers group. The self and spouse group was less likely to report a recent mammogram and clinical breast exam. The self only group did not differ significantly from the both nonsmokers group on any of the cancer screening measures. Results suggest that smoking status of a spouse may be an important correlate of women's cancer screenings. PMID- 10815699 TI - Male pattern baldness and clinical prostate cancer in the epidemiologic follow-up of the first National Health and Nutrition Examination Survey. AB - Male pattern baldness (MPB) and prostate cancer are common in American males; however, MPB is clinically observable decades earlier. Aging, androgens, and heritability are risk factors for both conditions. We prospectively studied the association between MPB and clinical prostate cancer in a cohort representative of the United States male population. A total of 4,421 men 25-75 years old without a history of prostate cancer were examined for baldness in the Epidemiologic Follow-up Study of the first National Health and Nutrition Examination Survey. Participants were followed from baseline (1971-1974) through 1992. Incident cases of prostate cancer were identified by interviews, medical records, and death certificates. Age-standardized incidence rates and proportional hazards models were used to examine the association between MPB and clinical prostate cancer. Prostate cancer was diagnosed in 214 subjects over 17 21 years of follow-up. The age-standardized incidence of prostate cancer was greater among men with baldness at baseline (17.5 versus 12.5 per 10,000 person years). The adjusted relative risk for prostate cancer among men with baldness was 1.50 (95% confidence interval, 1.12-2.00) and was similar regardless of the severity of baldness at baseline and was independent of other risk factors, including race and age. MPB seems to be a risk factor for clinical prostate cancer. PMID- 10815698 TI - Serum dehydroepiandrosterone and dehydroepiandrosterone sulfate and the subsequent risk of developing colon cancer. AB - This purpose of this study was to evaluate whether serum dehydroepiandrosterone (DHEA) and its sulfate conjugate, dehydroepiandrosterone sulfate (DHEAS), are associated with the likelihood of developing colon cancer. A nested case-control study was conducted using the serum bank and cancer registry in Washington County, Maryland. From a population of 20,305 county residents who donated blood in 1974, incident cases of colon cancer that occurred from 1975 to 1991 (n = 117) were matched to one cancer-free control by age, race, and sex. Serum specimens that were stored at -70 degrees C since 1974 were assayed for DHEA and DHEAS. Compared with the controls, the mean serum concentrations of cases were 3% lower for DHEA (P = 0.90) and 13% lower for DHEAS (P = 0.60). When DHEA levels were analyzed according to fourths, no noteworthy associations were observed. Compared with the lowest fourth, the highest fourth of serum DHEAS was nonsignificantly associated with a halving in the risk of colon cancer (odds ratio, 0.50; 95% confidence limits, 0.18, 1.37; Ptrend = 0.22), and further analyses showed the potential protective association was confined largely to males (highest-versus lowest fourth odds ratio, 0.26; 95% confidence limits, 0.06, 1.16; Ptrend = 0.06). This prospective study does not provide strong evidence that circulating DHEA and DHEAS concentrations are associated with the risk of colon cancer. Among men, DHEAS was associated with a decreased risk of colon cancer, but the association was within the bounds of chance. Further studies are needed to either support or refute the potentially promising lead hinted at by the results for DHEAS. PMID- 10815700 TI - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine induces a higher number of aberrant crypt foci in Fischer 344 (rapid) than in Wistar Kyoto (slow) acetylator inbred rats. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the most abundant heterocyclic amine carcinogen in the human diet and is a colon carcinogen in the rat. N-Acetyltransferase-2 (NAT2) catalyzes the conversion of PhIP and other heterocyclic amines to a DNA-reactive form. NAT2 has a polymorphic distribution in humans and other mammals, including rats. The rapid NAT2 genotype has been shown to be associated with increased colorectal cancer risk in some, but not all, human epidemiological studies. This investigation was designed to study the role of acetylator genotype in PhIP-induced colon carcinogenesis using aberrant crypt foci (ACF) as an intermediate biomarker. Five-week-old male, rapid acetylator Fischer 344 (F344) rats and slow-acetylator Wistar-Kyoto (WKY) rats were fed the semipurified AIN76A diet with 0.01% PhIP, 0.04% PhIP, or no PhIP (control) for 8 weeks. PhIP induced ACF in both rapid- and slow-acetylator rats; 0.04% PhIP induced more ACF than 0.01% PhIP. There was no difference in the number of ACF between rapid- and slow-acetylator rats that were fed 0.01% PhIP. However, 0.04% PhIP induced 2-fold higher ACF and a greater dose-dependent increase in PhIP-induced ACF in the rapid-acetylator F344 rats compared with the slow-acetylator WKY rats. The results support human epidemiological studies showing higher risk for colorectal cancer in rapid acetylators who frequently consume meat that is very well done. PMID- 10815701 TI - Correspondence re: L. R. Kidd et al., Urinary excretion of 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP) in white, African-American, and Asian American men in Los Angeles County. Cancer Epidemiol. Biomark. Prev., 8: 439-445, 1999. PMID- 10815702 TI - Correspondence re: S. A. Khan et al., The normal breast epithelium of women with breast cancer displays an aberrant response to estradiol. Cancer Epidemiol. Biomark. Prev., 8: 867-872, 1999. PMID- 10815703 TI - Semantic priming in patients with Alzheimer and semantic dementia. AB - Semantic priming for pairs of words having semantic relation (members of the same category) but not associative relation was tested in four patients with mild Alzheimer-type dementia (AD), three patients with semantic dementia (SD) and three normal controls. Priming effects were exhibited by all the AD patients and normal controls but by none of the SD patients. The facilitation did not differ depending on whether words were written in kanji or kana. The nature of semantic deficits in AD and SD is discussed. PMID- 10815704 TI - How patients with Parkinson's disease retrieve and manage cognitive event knowledge. AB - Several studies have pointed out that basal ganglia are involved in adaptive control of action at both motor and cognitive level. This study aimed to investigate how basal ganglia retrieve and manage script event knowledge required in planning behavior. Script event knowledge was investigated in patients with Parkinson's Disease using three kinds of activity that differed in familiarity. Unlike patients with prefrontal lesions, patients with Parkinson's Disease were able to order events in a typical sequence and obeyed the boundaries and hierarchies between events. In contrast, patients with Parkinson's Disease were impaired in evaluating how important each script event was within the context of goal-oriented planning activity. Our findings indicate that the prefrontal cortex and the basal ganglia are differentially involved in planning. The role of the basal ganglia might consist in providing a feedback about the goodness of each action while building up meaningful sequences of events during learning. PMID- 10815705 TI - Developmental dissociation between visual and auditory repetition priming: the role of input lexicons. AB - Contrasting theories posit the source of verbal repetition priming in the activation of preexisting memory representations in the input lexicons or, alternatively, in the formation of new episodic memory traces. The two hypotheses predict different outcomes from the comparison of developmental rates of visual and auditory verbal repetition priming. The activation theory predicts a developmental dissociation between the early maturation of auditory priming and the later maturation of visuo-verbal priming, contingent upon the discrepant acquisition rates of the auditory and visual input lexicons. The episodic theory, instead, does not make such an assumption. We administered visual and auditory implicit Stem Completion to 40 reading beginners (first-graders), 40 third graders and 20 fifth-graders. Consistent with previous reports, auditory priming was stable across different age groups. Visual priming and a measure of lexicality in reading, instead, showed a parallel developmental increase passing from reading beginners to third-graders and to fifth-graders. In the overall group, visual priming and the measure of lexicality in reading were significantly associated. These data describe a new developmental dissociation in the memory abilities of normal children and provide further support for the hypothesis that repetition priming for words reflects facilitated access to previously established memory representations. PMID- 10815706 TI - New learning and remote memory in the same and different domains of experience: implications for normal memory and amnesia. AB - This study investigated the correlation between new and remote memory ability in normal young subjects and evaluated whether using tests that tapped different domains of experience had any effect on the relationship between new and remote memory. Remote memory was assessed using a famous face test and new memory was evaluated using both novel faces and words. The main finding was a significant correlation between New and Remote face familiarity scores. A correlation was obtained when both tests tapped the same domain of memory, whereas when tests tapped different domains, there was no Remote-New correlation. Remote-New correlations may only exist between tests tapping the same domain of memory. This has implications for the interpretation of previous amnesic literature and for future investigations. PMID- 10815707 TI - Deficient intentional access to semantic knowledge in patients with severe closed head injury. AB - Patients after severe closed-head injury (CHI) demonstrate reduced ability to spontaneously utilize semantic memory during word-list memory tests and when requested to answer questions regarding general knowledge. However, they show normal lexical facilitation in both automatic and intentional semantic priming paradigms. The present study was aimed at investigating two alternative hypotheses a) that the deficit in semantic processing after CHI is the result of impaired access to an otherwise normal semantic system or b) that it reflects a loss of knowledge from a deteriorated semantic store. For this purpose, the performance of 15 CHI patients on an automatic Semantic Priming paradigm and on tests of Picture Naming and Semantic Judgment were compared to those of 14 normal controls (NC). Although CHI patients' reaction times were significantly slower than those of NCs, the semantic priming effect was comparable in the two groups. Instead, CHI patients performed significantly worse than NCs in the naming and semantic judgment tasks. These results provide evidence that CHI patients access semantic memory automatically at a normal rate. However, when the task is more demanding in terms of processing requests, then CHI patients' performance becomes defective. PMID- 10815709 TI - Verbal learning strategies in patients with multiple sclerosis. AB - Word list learning was studied in patients with a definite diagnosis of Multiple Sclerosis and in Normal Control subjects by means of the selective reminding procedure of Buschke and Fuld in two learning conditions: (1) using unrelated items and (2) paired-associate items. The Multiple Sclerosis patients displayed poor learning in both conditions. To identify the functional locus of their deficit, stochastic Markov chain analyses were performed, which allowed individual measurements of encoding, automatic and intentional retrieval abilities. On both tasks, encoding on the first trial and automatic retrieval on the subsequent trials were impaired in Multiple Sclerosis patients, whereas intentional retrieval, both with and without reminding by the examiner, appeared to be preserved. As all of the impaired abilities require a normal speed of information processing, the salient learning deficit of the Multiple Sclerosis patients could be tentatively traced back to the slowing down of their mental activity. PMID- 10815708 TI - Absence of amusia and preserved naming of musical instruments in an aphasic composer. AB - M.M., a right-handed, 74 year old professional musician and composer, presented with a progressive aphasia with a severe anomia. His musical competence was apparently totally preserved, and he continued his activity as a composer. There was a striking discrepancy between his impaired naming of nonmusical stimuli and his normal naming of musical instruments' sounds. We suggest that the preservation of skills in the musical domain results from an expanded cortical representation of this function in the left hemisphere, secondary to his lifelong formal training, and to the high level of his professional competence. As for his preserved naming of musical instruments, we argue that the early age-of acquisition and higher than "normal" frequency/familiarity for names of musical instruments facilitate the access to their lexical representation and/or their retrieval within the lexicon. PMID- 10815710 TI - Acalculia, aphasia and spatial disorders in left and right brain-damaged patients. AB - The paper reports the performance of 50 left- and 26 vascular right-brain-damaged (LBD, RBD) patients in the EC301 Calculation Battery, which explores different aspects of number and calculation processing. All patients underwent a comprehensive neuropsychological testing that also included evaluation for the presence and type of aphasia in LBD patients, and of spatial disorders in RBD patients. LBD were subdivided in three groups: non-aphasic (NA), Broca and Wernicke aphasics. Results indicate that language and calculation disorders can dissociate. The relationship between spatial and calculation disorders in RBD patients is less clear. No significant difference was found between Broca and Wernicke aphasics, nor between NA and RBD patients. In the transcoding tasks (reading or writing to dictation numbers and number words, for instance) syntactic errors were the most frequent type of errors in all groups. They were also present when neither the input nor the required response was in the Arabic code, and a word-by-word strategy could have been used to read the number word or write a spoken number in the orthographic code. PMID- 10815711 TI - Speech timing in Thai left- and right-hemisphere-damaged individuals. AB - An acoustic analysis of syllable duration in short Thai phrases was conducted to evaluate the effects of focal brain damage on the control of speech timing. Almost all 35 of the subjects had participated in each of four previous companion studies: 13 left-brain-damaged (6 nonfluent aphasics; 7 fluent aphasics), 14 right-brain-damaged patients, and 8 normal controls. Somewhat surprisingly, results revealed relatively normal timing patterns in 3-syllable phrases in all subject groups. A comparison of the current study and the four others, however, led us to conclude that Thai-speaking nonfluent aphasics exhibit a speech timing deficit regardless of the linguistic level of representation, whereas timing deficits in fluent aphasics appear to be restricted to units larger than a syllable. Speech timing, on the other hand, appears to be intact across the board in right-brain-damaged individuals. Findings are brought to bear on theories of temporal control in brain-damaged patients. PMID- 10815712 TI - Improving the clinical diagnosis of personal neglect: a reformulated comb and razor test. AB - Beschin and Robertson (1997) devised a simple clinical test of left personal neglect, which characterises personal grooming behaviour according to the proportion of the total activity that is directed to the left side of the body. Although this test proved highly reliable, and more sensitive than prior diagnostic techniques, its formulation may yet be improved. The present paper reports a reanalysis of Beschin and Robertson's (1997) data, using additional control subjects, and a formula which characterises personal neglect as a lateral bias of behaviour rather than as a lateralised deficit. It is shown that this formula greatly enhances the test's sensitivity to the behavioural abnormalities of brain damaged patients, and it is recommended that this modification be adopted for the future diagnosis of personal neglect. PMID- 10815713 TI - Reading latency of words and nonwords in Alzheimer's patients. AB - Contrasting data on reading ability in Alzheimer's disease patients have been reported in the literature. Recently Patterson, Graham and Hodges (1994) found that irregular words were misread by demented subjects, while regular words were read correctly. The present study hypothesizes that reading latency may be a sensitive measure of Alzheimer's patients reading impairment. Fifteen Alzheimer's patients were compared with 17 elderly normal subjects on three tasks that used the same set of concrete, regular words: a picture naming task, a word-picture matching task and a word-nonword reading task. The results of the study indicate that reading latency is longer in Alzheimer's patients than in normal subjects, and that misnamed and mismatched words are read with the same mechanism as nonwords. PMID- 10815714 TI - Basic concepts of artificial neural network (ANN) modeling and its application in pharmaceutical research. AB - Artificial neural networks (ANNs) are biologically inspired computer programs designed to simulate the way in which the human brain processes information. ANNs gather their knowledge by detecting the patterns and relationships in data and learn (or are trained) through experience, not from programming. An ANN is formed from hundreds of single units, artificial neurons or processing elements (PE), connected with coefficients (weights), which constitute the neural structure and are organised in layers. The power of neural computations comes from connecting neurons in a network. Each PE has weighted inputs, transfer function and one output. The behavior of a neural network is determined by the transfer functions of its neurons, by the learning rule, and by the architecture itself. The weights are the adjustable parameters and, in that sense, a neural network is a parameterized system. The weighed sum of the inputs constitutes the activation of the neuron. The activation signal is passed through transfer function to produce a single output of the neuron. Transfer function introduces non-linearity to the network. During training, the inter-unit connections are optimized until the error in predictions is minimized and the network reaches the specified level of accuracy. Once the network is trained and tested it can be given new input information to predict the output. Many types of neural networks have been designed already and new ones are invented every week but all can be described by the transfer functions of their neurons, by the learning rule, and by the connection formula. ANN represents a promising modeling technique, especially for data sets having non-linear relationships which are frequently encountered in pharmaceutical processes. In terms of model specification, artificial neural networks require no knowledge of the data source but, since they often contain many weights that must be estimated, they require large training sets. In addition, ANNs can combine and incorporate both literature-based and experimental data to solve problems. The various applications of ANNs can be summarised into classification or pattern recognition, prediction and modeling. Supervised 'associating networks can be applied in pharmaceutical fields as an alternative to conventional response surface methodology. Unsupervised feature-extracting networks represent an alternative to principal component analysis. Non-adaptive unsupervised networks are able to reconstruct their patterns when presented with noisy samples and can be used for image recognition. The potential applications of ANN methodology in the pharmaceutical sciences range from interpretation of analytical data, drug and dosage form design through biopharmacy to clinical pharmacy. PMID- 10815715 TI - Fluorimetric determination of some thioxanthene derivatives in dosage forms and biological fluids. AB - A simple and highly sensitive method is proposed for the fluorimetric determination of four thioxanthene derivatives, namely: chlorprothixene, clopenthixol, flupentixol and thiothixene, in dosage forms and biological fluids. The method involves the use of nitrous acid as an oxidant to produce the corresponding fluorescent thioxanthenone sulphoxides. The experimental parameters were carefully studied and incorporated into the procedures. The results obtained compare favourably with those obtained by the official methods. The concentration fluorescence plots were rectilinear over the range of 0.04-0.4 microg/ml for thiothixene, and 0.02-0.25 microg/ml for the other compounds, with minimum detectability (S/N = 2) of 2 ng/ml for all the studied compounds except thiothixene which was 4 ng/ml. The proposed method was applied to the determination of the studied compounds in dosage forms. The results obtained were in good agreement with those obtained adopting the USP XIII method. The proposed method was further applied to the determination of flupentixol in spiked human urine and plasma, the percentage recoveries were 94.39 +/- 1.81 and 96.46 +/- 0.28, respectively. A proposal of the reaction pathway was presented. PMID- 10815716 TI - RP-LC method for the determination of cetirizine in serum. AB - The development and evaluation of HPLC method for quantifying cetirizine in human serum is described. The method involves liquid phase extraction of cetirizine in methylene chloride, adding diazepam as an internal standard, followed by separation on a reversed phase C18 Novapak column (150 x 3.9 nm; 4 microm), and employing a UV-detection set at 230 nm at ambient temperature. The mobile phase consists of a 13 mM phosphoric acid solution and acetonitrile (61:39 v/v) adjusted to pH 2.8 with 5 M NaOH. The assay is linear from 10 to 500 ng ml(-1) with a detection limit of 5 ng ml(-1) and a mean recovery of 96.5%. The applicability of this method in pharmacokinetic studies is evaluated. PMID- 10815717 TI - Stability study of selected adenosine nucleosides using LC and LC/MS analyses. AB - The stability of the naturally occurring nucleoside, adenosine, and two synthetic chlorine-containing analogues, 2-chloroadenosine and 5'-chloro-5'-deoxyadenosine was studied using high performance liquid chromatography (LC) and liquid chromatography in combination with mass spectrometry (LC/MS). The stability of the examined nucleosides over pH range of 2-10 and at temperatures 40, 60 and 80 degrees C was measured using an LC method, whereas the products of hydrolysis were identified using LC/MS. The LC data indicated that the hydrolysis of the nucleosides followed pseudo-first order kinetics. The MS data proved that the fragment ions at m/z 136.3 and 170.3 referred to the hydrolytic products, adenine and 2-chloroadenine, respectively. The calculated values of the hydrolysis rate constant and half-life indicated that the presence of chlorine atom in the nucleoside base moiety increases apparently the stability of 2-chloroadenosine against acid hydrolysis compared to 5'-chloro-5'-deoxyadenosine and adenosine. PMID- 10815718 TI - Adsorption of endotoxin on glass in the presence of rhIL-11. AB - Poor recovery of spiked endotoxin in the Limulus amebocyte lysate assay (LAL assay) was observed in the presence of recombinant human interleukin-11 (rhIL 11), a cationic, hydrophobic protein. Detection of endotoxin activity remaining in the empty glass tubes in which endotoxin and rhIL-11 mixtures were incubated indicated adsorption of endotoxin on glass. At low concentrations of rhIL-11, a correlation between endotoxin adsorbed on glass and a decrease of endotoxin in solution was observed. Adsorption of rhIL-11 on glass correlated with adsorption of endotoxin, which indicates that rhIL-11 mediates adsorption of endotoxin on glass. Consequently, adsorption of endotoxin on glass may occur in the presence of other substances which bind to both of endotoxin and glass. PMID- 10815719 TI - The qualitative and quantitative determination of quinolones of first and second generation by capillary electrophoresis. AB - Capillary electrophoresis (CE) was applied to the study of 10 quinolones of first and second generation--nalidixic acid, oxolinic acid, pipemidic acid, cinoxacin, norfloxacin, ciprofloxacin, ofloxacin, pefloxacin, fleroxacin, and flumequine. Separation was performed on a fused silica capillary (75 microm-60 cm) using a phosphate buffer (pH 7.0, 125 mM). Detection was at 214 nm. Only norfloxacin and ciprofloxacin cannot be separated in this way. Because of the specificity of the method, the identification of the individual quinolones by their migration time was possible. The same system has been applied for the quantitative determination of quinolones in tablets and capsules. Excipients do not adversely affect the results. Some parameters (linearity, precision, accuracy) were validated. Especially the possibility of simultaneous quantification and identification of the active ingredient in the finished product is very attractive. PMID- 10815720 TI - Fluorescence determination of sulphobutylether-beta-cyclodextrin in human plasma by size exclusion chromatography with inclusion complex formation. AB - A selective method for the determination of sulphobutylether-beta-cyclodextrin (SBECD) in human plasma has been developed and validated over the range 4-200 microg ml(-1). SBECD is extracted from plasma using end-capped cyclohexyl solid phase extraction cartridges. This is followed by high performance size exclusion chromatography with a mobile phase consisting of 1-naphthol (0.1 mM) in methanol potassium nitrate (0.2 M) (1:9 v/v), 1 ml min(-1). The high aqueous content of the mobile phase quenches the fluorescence of 1-naphthol. However, the naphthol forms an inclusion complex with SBECD. The non-polar 'bucket' environment of the inclusion region restores the fluorescence, which is measured at excitation and emission wavelengths of 290 and 360 nm, respectively, when SBECD elutes from the column. PMID- 10815721 TI - Automated flow-injection spectrophotometric determination of catecholamines (epinephrine and isoproterenol) in pharmaceutical formulations based on ferrous complex formation. AB - A novel automated flow-injection spectrophotometric method for the determination of catecholamines (epinephrine and isoproterenol) has been developed based on the formation of their coloured complexes with Fe(II) in aminoacetic-carbonate buffer pH 8.3 and measuring of the absorbance peaks at the lambda(max) of 530 nm. A fully automated FIA system controlled by home-made software (FIA-MOD) was used for optimising the chemical and manifold parameters and running of routine measurements. The calibration graph was linear in the range of 5-200 mg l(-1) for epinephrine with an RSD of 0.24% (n = 5; c = 150 mg l(-1)) and 10-300 mg(-1) for isoproterenol with an RSD of 0.13% (n = 5; c = 200 mg l(-1)). Measurement throughput was 120 h(-1) ensuring a sample throughput of 40 h(-1) analysed in triplicate. Common excipients for tablets and injections were found not interfering. The proposed method was applied for the assay of various commercial pharmaceutical formulations containing epinephrine and isoproterenol and for the content uniformity test for the isoproterenol tablets. The assay results with RSD 2-4% (n = 3) were comparable with those obtained with the official USP XXIII methods (mean difference 1.9%). PMID- 10815722 TI - Determination of free and encapsulated oligonucleotides in liposome formulated drug product. AB - Liposomes have been recognized as new delivery vehicles for peptide and oligonucleotide drugs, offering effective drug protection and influencing drug distribution from the circulation to tissues. To ensure consistent formulation behavior and drug distribution, the amounts of free and encapsulated active pharmaceutical ingredient in the liposome formulation must be determined. A simple and reliable method has been developed for the determination of free and encapsulated oligonucleotide drugs in liposomes. Capillary electrophoresis in entangled polyacrylamide solution was optimized for the analysis. Liposome samples were treated to release encapsulated oligonucleotide so that total concentration could be determined, and untreated liposomes were applied directly to capillary column to determine the fraction of free oligonucleotides. Recoveries are 96-105% and relative standard deviations are generally 2-3%. Multiple liposome types were analyzed with satisfactory results. PMID- 10815723 TI - Stability and enzymatic hydrolysis of quaternary ammonium-linked glucuronide metabolites of drugs with an aliphatic tertiary amine-implications for analysis. AB - Quaternary ammonium-linked glucuronide (N+-glucuronide) metabolites formed at aliphatic tertiary amine functional groups of xenobiotics have not been previously systematically studied with respect to their stability over a wide pH range and the ease of enzymatic hydrolysis by beta-glucuronidase from various sources. Three and four N+-glucuronide metabolites were respectively studied regarding their non-enzymatic and enzymatic stabilities where the metabolites were quantified by HPLC procedures. The N+-glucuronide metabolites of clozapine, cyclizine, and doxepin were stored at 18-22 degrees C in buffers at each nominal pH value over the 1-11 pH range. All three metabolites were stable for 3 months over the 4-10 pH range, while two metabolites slowly degraded (k in the range 0.002-0.01 days(-1)) at each of the other extreme pH values. In the initial enzymatic study the N+-glucuronide metabolites of chlorpromazine, clozapine, cyclizine, and doxepin were each treated in pH 5.0 and 7.4 buffers at 37 degrees C with beta-glucuronidase from three different sources, namely commercial brands from bovine liver, mollusks (Helix pomatia), and bacteria (Escherichia coli). Clozapine N+-glucuronide and the standard phenolphthalein O-glucuronide were susceptible to hydrolysis by the enzyme from all three sources. In contrast, the other three N+-glucuronide metabolites were resistant to hydrolysis, except for the E. coli source of beta-glucuronidase at pH 7.4. Also when examined at 50-fold increase in concentration of the enzyme sources from bovine liver and H. pomatia cyclizine N+-glucuronide was still resistant to hydrolysis by the former enzyme preparation. The optimum pH for the hydrolysis of each of the four N+-glucuronide metabolites from the E. coli enzyme source was investigated and was found to be in the pH range 6.5-7.4. These data have important implications with respect to the analysis of N+-glucuronide metabolites formed at an aliphatic tertiary amine: in general, their non-enzymatic stability will not be an important factor in the development of an analytical procedure, and when developing an indirect approach to the analysis of N+-glucuronide metabolites that involves beta-glucuronidase hydrolysis to the aglycone preliminary work should involve determining the appropriate enzyme source, buffer pH, and length of time of incubation. PMID- 10815724 TI - Bioanalytical method validation design for the simultaneous quantitation of analytes that may undergo interconversion during analysis. AB - In the analysis of post-dose biological samples for quantitative determination of two analytes that can potentially undergo interconversion, it is essential to minimize the interconversion during the multiple steps of the bioanalytical method. However, even after optimizing the conditions of each step, some interconversion may be unavoidable. Even then, a method can be developed for the accurate simultaneous determination of the two analytes in post-dose biological samples if the composition, in terms of the ratio of the concentrations of the two analytes, of the calibration standards and quality control (QC) samples are selected judiciously, in relation to the composition of the unknown samples to be analyzed. As an example of such interconverting analytes, a delta-hydroxy acid compound (analyte 1) and its delta-lactone (analyte 2) were selected as model compounds that can potentially undergo interconversion. The effects of changing the relative concentrations of the two analytes in QC samples vis-a-vis the calibration standards on the performance of the method under conditions were investigated where: (a) the interconversion between the two analytes was minimized; (b) the conversion of analyte 2 to analyte 1 was enhanced; (c) the interconversion between the two analytes was enhanced. The results showed that the method performance, as measured by the accuracy and precision of the QC samples, was not acceptable when the ratio of concentration of analyte 1 to that of analyte 2 in the QC samples was different from that in the calibration standards and the conditions used facilitated the conversion of one analyte to the other. However, when the relative concentration of the two analytes in the QC samples was identical to that of the calibration standards, the method performance was acceptable under all three conditions of interconversion. This was because the same degree of interconversion took place in the QC samples and calibration standards. The purpose of QC samples in bioanalytical methods is to gauge how the method will perform for the analysis of post-dose test samples and hence, ideally, the relative concentrations of the analytes in QC samples, should be selected to mimic the anticipated concentrations in the test samples. However, the relative concentrations of the analytes in test samples may not be known a priori, or may change from sample to sample; therefore, it is not always possible to construct QC samples that exactly mimic the relative concentrations of analytes in the test samples. Thus, in order to cover the variety of test samples, the method should include, in addition to QC samples that contain the analytes at the same relative concentration as in the calibration standards, QC samples with relative concentrations that are different from those in the calibration standards, including those that contain only analyte 1 and only analyte 2. In addition, the conditions adopted for the method should favor the minimization of the conversion of the analyte that is expected to be the major component in the post-dose test samples. PMID- 10815725 TI - The use of near-infrared spectroscopy to monitor the mobility of water within the sarafloxacin crystal lattice. AB - Near-infrared spectroscopy (NIRS) is proposed as a technique to study the mobility of water within the sarafloxacin crystal lattice. An investigation of two samples of sarafloxacin revealed that NIRS can distinguish between acceptable and unacceptable batches for formulation purposes. X-ray powder diffraction (XRPD), mid-infrared (mid-IR) spectroscopy, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) could not detect any differences between the two samples. NIRS detected differences in the location or orientation of the water molecules within the crystal lattices. PMID- 10815726 TI - Rapid and sensitive ethidium bromide fluorescence quenching assay of polyamine conjugate-DNA interactions for the analysis of lipoplex formation in gene therapy. AB - A rapid and sensitive fluorescent assay method is reported for assessing polyamine conjugate calf thymus DNA binding affinity using cholesterol polyamine carbamates with ethidium bromide as a probe. A reproducible method has been developed with an optimal excitation wavelength. Salt concentration is shown to be a critical parameter for both the observed fluorescence intensity of ethidium intercalated in DNA, and also for the binding of positively charged polyammonium ions to DNA, effecting charge neutralisation. This charge neutralisation precedes DNA condensation, a key first step in gene therapy. PMID- 10815727 TI - Mixed-mechanism ionization to enhance sensitivity in atmospheric pressure ionization LC/MS. AB - A novel dual-mechanism ionization technique for LC/MS/MS has been observed, characterized and applied to the quantitation of a tertiary amine-containing drug compound in dog plasma. This mixed-mechanism ionization approach can improve the sensitivity of the pneumatically assisted electrospray experiment. Under conditions of higher than normal chromatographic flow and lower electrospray voltage, approximately a 4-fold increase in sensitivity was realized. A detection limit of 16 pg (45 fmol) on-column, and inter-day imprecision and inaccuracy of < 11 and < 15%, respectively, were obtained. A trade-off in concentration sensitivity in favor of ease of sample preparation was made to increase sample throughput. Although results strongly suggest that mixed-mechanism ionization is in operation, and that pneumatically assisted electrospray is a partial contributor to the overall ionization process, the exact nature of the second mechanism of ionization is unclear at this time. PMID- 10815728 TI - Quantitation of N-(3,5-dichloropyrid-4-yl)-3-cyclopentyloxy-4-methoxybenzamide and 4-amino-3,5-dichloropyridine in rat and mouse plasma by LC/MS/MS. AB - The metabolism of N-(3,5-dichloropyrid-4-yl)-3-cyclopentyloxy-4-methoxybenzamide (RP73401), a phosphodiesterase IV (PDE IV) inhibitor is extensive (unpublished); however, until recently, studies for this compound did not report 4-amino-3,5 dichloropyridine (ADCP) as a metabolite either in vitro or in vivo. This prompted a reinvestigation into the metabolism of RP73401 in rats and mice using mass spectrometry. The results of the reinvestigation confirmed that 4-amino-3,5 dichloropyridine was formed via the metabolism of RP73401 both in vitro and in vivo. In order to further investigate RP73401 hydrolysis in vivo, a liquid chromatography/mass spectrometry assay was developed and validated for the simultaneous determination of RP73401 and ADCP in rat and mouse plasma. The method used Waters Oasis HLB brand solid phase extraction cartridges to isolate the analytes (RP73401 and ADCP) and internal standard from the plasma. HPLC chromatographic separation was achieved using a Zorbax SB C18 HPLC column and detection was accomplished using positive ion atmospheric pressure chemical ionization tandem mass spectroscopy in multiple reaction monitoring (MRM) mode. The assay was developed and validated over the range of 0.5-100 ng ml(-1) for RP73401 and 5-500 ng ml(-1) for ADCP using 0.050 ml of plasma. The assay proved to be sensitive, accurate, precise and specific for RP73401 and ADCP. Intraday and interday quality control results routinely showed accuracy and precision to be within +/- 20%. This LC/MS/MS method was subsequently employed to investigate the hydrolysis of RP73401 in the rat and mouse, and determine the effects of tri o-tolyl phosphate (TOTP, a carboxylesterase inhibitor) preadministration on the hydrolysis reaction in the rat. PMID- 10815729 TI - Application of micellar electrokinetic chromatography for the separation of retinoids. AB - The applicability of micellar electrokinetic chromatography (MEKC) using sodium dodecylsulphate (SDS) as pseudo-stationary phase for the separation of five retinoids (retinol, retinal, retinyl acetate, retinyl palmitate, retinoic acid), was investigated. The effects of the acetonitrile content, the SDS concentration, the pH and the addition of Brij 35 to the background electrolyte on the migration behaviour of the retinoids were determined. It was found that the effective mobilities of retinol, retinal and retinyl acetate could be easily regulated through the ACN content and the SDS concentration of the BGE. The electrophoretic behaviour of the very hydrophobic retinyl palmitate was abnormal. Under various conditions this compound showed up as a late, very sharp peak. A strong indication was found that the retinyl palmitate forms a stable, charged complex with SDS during sample preparation. The mobility of the retinyl palmitate peak could be regulated, independently from the other peaks, through the Brij concentration of the BGE. Using a running buffer consisting of Tris buffer (pH 8), 20 mmol l(-1) SDS, 1 mmol l(-1) Brij 35 and 35% (v/v) acetonitrile, a complete separation of the five retinoids could be realised in less than 20 min. PMID- 10815731 TI - The origins and precolonial epidemiology of tuberculosis in the Americas: can we figure them out? AB - Paleologic evidence of tuberculosis in the precolonial Americas is reviewed to cast light on its origins and subsequent epidemiology. The genus Mycobacterium is an ancient one, and M. tuberculosis may have differentiated 20,400 to 15,300 years ago. The Americas were peopled by migrants from Asia in two major migrations, one occurring more than 20,000 years ago and the other 12,000 to 11,000 years ago. Tuberculosis reached the Americas with these migrants, persisting at a low level of endemnicity in small, dispersed population groups. Beginning about 1500 years ago, an epidemic of tuberculosis began, probably in the Andean region of South America. It did not reach or subsided in time to leave highly susceptible indigenous American populations at the time of European colonization. PMID- 10815730 TI - Redefining MDR-TB transmission 'hot spots'. AB - Halting further spread of multidrug-resistant tuberculosis (MDR-TB) requires both new resources and a renewed discussion of priority setting informed by estimates of the existing burden of this disease. The 1997 report of the first phase of the global survey by the World Health Organization (WHO) and the International Union Against Tuberculosis and Lung Disease (IUATLD) uses the indicator of the proportion of TB cases that are MDR-TB to identify MDR-TB 'hot spots'. We sought to refine the definition of MDR-TB transmission 'hot spots'. For this purpose, we obtained estimates of two additional indicators for regions where data are available: MDR-TB incidence per 100,000 population per year, and expected numbers of new patients with MDR-TB per year. There is generally much agreement in the three indicators considered, and some differences also appear. We conclude that it is useful, when defining indicators of MDR-TB transmission 'hot spots', to include estimates of underlying TB incidence rates and of absolute numbers of MDR TB cases. Estimating the force of morbidity of MDR-TB in a population is important for comparing this burden across settings with very different underlying TB incidence rates; estimating the absolute number of MDR-TB patients will be critical for planning the delivery of directly observed MDR-TB therapy and the rational procurement of second-line drugs. Through this exercise, we aim to initiate discussion about improved methods of quantifying and comparing current MDR-TB transmission 'hot spots' that require intervention. PMID- 10815732 TI - Community involvement in tuberculosis control: lessons from other health care programmes. AB - Decentralising tuberculosis control measures beyond health facilities by harnessing the contribution of the community could increase access to effective tuberculosis care. This review of community-based health care initiatives in developing countries gives examples of the lessons for community contribution to tuberculosis control learned from health care programmes. Sources of information were Medline and Popline databases and discussions with community health experts. Barriers to success in tuberculosis control stem from biomedical, social and political factors. Lessons are relevant to the issues of limited awareness of tuberculosis and the benefits of treatment, stigma, restricted access to drugs, case-finding and motivation to continue treatment. The experience of other programmes suggests potential for an expansion of both formal and informal community involvement in tuberculosis control. Informal community involvement includes delivery of messages to encourage tuberculosis suspects to come forward for treatment and established tuberculosis patients to continue treatment. A wide range of community members provide psychological and logistic support to patients to complete their treatment. Lessons from formal community involvement indicate that programmes should focus on ensuring that treatment is accessible. This activity could be combined with a variety of complementary activities: disseminating messages to increase awareness and promote adherence, tracing patients who interrupt treatment, recognising adverse effects, and case detection. Programmes should generally take heed of existing political and cultural structures in planning community-based tuberculosis control programmes. Political support, the support of health professionals and the community are vital, and planning must involve or stem from the patients themselves. PMID- 10815733 TI - DOT or not? Direct observation of anti-tuberculosis treatment and patient outcomes, Kerala State, India. AB - SETTING: The Pathanamthittha District of Kerala State, India, where the directly observed treatment, short-course (DOTS) programme was started in October 1994. OBJECTIVE: To determine the frequency with which direct observation actually occurred within a district-level DOTS programme, and the association of treatment observation with treatment outcome under programme conditions. DESIGN: This retrospective study included 200 consecutive, newly-detected, smear-positive patients registered under the project between February 1995 and February 1996 at the District Tuberculosis Centre, as well as health workers responsible for providing directly observed treatment (DOT) who were separately and confidentially interviewed. Treatment outcomes were identified from results of sputum smear examinations for acid-fast bacilli. RESULTS: Although all patients were recorded as having received DOT, more than a quarter of patients (26.5%) did not actually receive it. The 53 patients who were not directly observed were much more likely to have treatment failure or relapse, as compared to those who had received DOT (45% vs 3%, relative risk 16.6, 95% confidence intervals 6-46, P < 0.001). Women were somewhat less likely than men (61% vs 76%, P = 0.06) to receive DOT. Non-receivers of DOT accounted for 86% (24/28) of treatment failures or relapses. CONCLUSION: Patients treated without direct observation have a substantially higher risk of adverse outcome than those treated under direct observation. To be maximally effective, the DOTS programme must be both confidential and convenient. PMID- 10815734 TI - Tuberculosis during infancy. AB - SETTING: A worldwide re-emergence of tuberculosis has been observed during the last decade. However, few studies of infants with tuberculosis appear in the literature. OBJECTIVE: To describe tuberculosis during infancy. DESIGN: The records of all infants diagnosed with tuberculosis at a tertiary care hospital from 1982 to 1998 were reviewed. RESULTS: Thirty-nine infants with a median age of 10 months were identified, 59% of whom presented during the second half of the study period. Diagnoses included endothoracic tuberculosis (33 patients), meningitis (3), miliary tuberculosis (2) and cervical lymphadenitis (1). Reasons for medical evaluation were the onset of symptoms (25 patients), contact investigation (12) and tuberculin skin test screening (2). Common signs and symptoms included fever (22 patients), cough (7), appetite loss (4) and wheezing/rales (4). Chest X-ray revealed hilar adenopathy (22 patients), infiltrates (16), atelectases (3) and miliary pattern (2). Cultures were attempted in nine patients and were positive in seven. All patients responded promptly to treatment. No complications or deaths occurred. CONCLUSION: Tuberculosis in infants has been diagnosed increasingly during the last decade. Endothoracic tuberculosis predominates. One third of the patients were diagnosed due to contact investigation. As early diagnosis and treatment appears to prevent complications and reduce mortality, pediatricians should be alert for tuberculosis in an infant with an atypical picture suggestive of infection. PMID- 10815735 TI - Medical students at risk of nosocomial transmission of Mycobacterium tuberculosis. AB - SETTING: University and teaching hospital in Rio de Janeiro, Brazil, a city with a high prevalence of tuberculosis (TB). OBJECTIVE: To determine whether medical students are at increased risk of nosocomial transmission of Mycobacterium tuberculosis relative to other university students. DESIGN: A cross-sectional study of medical and chemical engineering students in different levels of their training programmes. Information about socio-demographic characteristics, BCG vaccination history, and potential exposures to TB were obtained using a standardised questionnaire. Tuberculin skin testing (TST) was used to determine the prevalence of infection with TB. RESULTS: Medical students have an increasing prevalence of TST positivity as they advance in their training programme to increasing levels of study (4.6%, 7.8%, 16.2%, respectively, P < 0.001), but chemical engineering students do not (4.2%, 4.3%, 4.4%, respectively, P = 0.913). The risks are greatest during the years of clinical training, when medical students have increased contact with patients. CONCLUSIONS: Medical students in this setting may be at increased risk of M. tuberculosis infection, relative to chemical engineering students. A programme of routine tuberculin skin testing is needed, combined with interventions to reduce the risk of nosocomial transmission in the workplace. PMID- 10815736 TI - Acquired anti-tuberculosis drug resistance in Yaounde, Cameroon. AB - SETTING: Tuberculosis centre of Hopital Jamot, Yaounde, Cameroon. OBJECTIVES: To determine the prevalence of acquired resistance (ADR) to the main anti tuberculosis drugs, and to identify risk factors associated with its occurrence in Yaounde. DESIGN: A total of 111 previously treated adults admitted consecutively to the tuberculosis centre with sputum smear-positive pulmonary tuberculosis between June 1996 and July 1997 were included in the study. Information on potential risk factors for ADR was obtained from each patient, and human immunodeficiency virus (HIV) serostatus was determined. Drug susceptibility testing to the main anti-tuberculosis drugs was performed on cultures of Mycobacterium tuberculosis complex isolated from sputum samples of each patient by the indirect proportion method. All patients whose isolates tested resistant to at least one anti-tuberculosis drug were defined as having ADR. RESULTS: Growth of M. tuberculosis complex was obtained from sputum specimens of 98 (88.3%) of the 111 patients studied; 57 (58.2%) of these were resistant to at least one anti-tuberculosis drug. Resistance to isoniazid was the most common (54.1%), followed by resistance to rifampicin (27.6%), streptomycin (25.5%) and ethambutol (12.2%). Multidrug resistance was observed in 27 (27.6%) of the cases. In a multivariate logistic regression analysis, ADR was significantly associated only with monotherapy use in previous tuberculosis treatment(s) (P = 0.03). CONCLUSION: The rate of ADR of M. tuberculosis is quite high in Yaounde. Acquired resistance to rifampicin alone or in combination with isoniazid is also high. Monotherapy in previous anti-tuberculosis treatment(s) is a significant predictor of ADR in previously treated patients in Yaounde. These results underscore the urgent need for the re-establishment of a tuberculosis control programme, using the DOTS strategy, in Cameroon. PMID- 10815737 TI - Drug-resistant tuberculosis in South African gold miners: incidence and associated factors. AB - SETTING: A gold mining company in the Free State Province of South Africa. OBJECTIVE: To document the incidence of and factors associated with drug resistant tuberculosis (TB) in South African gold miners. DESIGN: Review of Mycobacterium tuberculosis drug susceptibility records for the period from 1 July 1993 to 30 June 1997. RESULTS: Over the study period, 2241 miners had culture positive M. tuberculosis pulmonary disease where isolates were tested for drug susceptibility to the four primary anti-tuberculosis drugs. The proportions of primary and acquired drug resistance were respectively 7.3% and 14.3% for isoniazid and 1.0% and 2.8% for resistance to at least isoniazid and rifampicin (multidrug resistance). Resistance to streptomycin and ethambutol was uncommon, and rifampicin monoresistance was rare. No significant factors for primary drug resistance were identified. Patients with retreatment pulmonary TB who failed primary TB treatment (versus cure) were significantly more likely to have TB with resistance to any TB drug or MDR (odds ratios respectively 9.82, 95%CI 2.97-33.5, and 18.74, 95%CI 1.76-475). Human immunodeficiency virus (HIV) infection was not significantly associated with primary or acquired drug resistance, and there was no trend of increasing resistance over time. CONCLUSION: Anti-tuberculosis drug resistance has remained stable despite the HIV epidemic and increasing TB rates. Directly observed therapy may have contributed to containing the level of drug resistance. Adherence to and completion of treatment are essential to prevent drug resistance and treatment failure, including in situations with high HIV prevalence. PMID- 10815738 TI - Exclusive mutations related to isoniazid and ethionamide resistance among Mycobacterium tuberculosis isolates from Korea. AB - SETTING: The single base change at the 94th codon of inhA has been referred to as the event that confers resistance on the drugs isoniazid (INH) and ethionamide (ETH) in Mycobacterium smegmatis and M. bovis. From this observation, it has been anticipated that some of the INH-resistant clinical isolates of M. tuberculosis would carry missense mutations in the same region of the gene. However, few polymorphisms have been identified in this region among INH-resistant isolates. OBJECTIVE: To understand the molecular basis for M. tuberculosis resistance to INH and ETH. DESIGN: The sequence polymorphism at the 94th codon of inhA among M. tuberculosis isolates from Korea was analyzed by polymerase chain reaction (PCR) cloning and sequence analysis. RESULTS: No nucleotide change at the 94th codon of inhA was detected in any of the 24 INH-resistant isolates analyzed in this study. On the other hand, a point mutation was found exclusively at the regulatory region flanking a putative ribosome-binding site of the inhA locus in 14 isolates. Interestingly, all the mutations were of the same kind, which substitutes C to T. Among 14 isolates, 12 were resistant to INH as well as to ETH, while two were resistant to INH only. DISCUSSION: It seems that mutations previously found at the 94th codon of inhA have no particular relationship with the mechanism involved in the resistance of M. tuberculosis to INH and/or ETH. On the other hand, the resistance mechanism of M. tuberculosis to INH/ETH may involve an altered level of InhA, an expression which may have been influenced by the sequence change in the regulatory region of the inhA locus. PMID- 10815739 TI - HIV-1 co-infection in children hospitalised with tuberculosis in South Africa. AB - SETTING: Hospitals associated with the Department of Paediatrics at the University of the Witwatersrand, Johannesburg, South Africa. OBJECTIVES: To define the prevalence of human immunodeficiency virus (HIV) co-infection and differences in clinical presentation between HIV-infected and non-infected hospitalised children with tuberculosis. DESIGN: Children were prospectively enrolled between August 1996 and January 1997. RESULTS: Of 161 children enrolled, 42% were HIV-infected, including 67/137 with pulmonary tuberculosis (PTB) and 1/24 with extra-pulmonary disease (EPTB). Positive microscopy or bacteriology did not differ by HIV status for children with either PTB or EPTB. Although age did not differ between HIV-infected and non-infected children with PTB, non-HIV infected children with EPTB were significantly older than those with PTB only (median age 32 months vs 14.5 months, P = 0.004). Chronic weight loss, malnutrition and the absence of BCG scarring were more common in HIV-infected children with PTB. HIV-infected children were also more likely to show cavitation (P = 0.001) and miliary TB (P = 0.01) on chest X-ray. Reactivity to tuberculin (> or = 5 mm and > or = 10 mm in HIV-infected and non-infected children, respectively) was significantly lower in HIV-infected children, as were CD4+ lymphocyte levels. The mortality rate during the study was 13.4% in HIV-infected children compared with 1.5% in non-HIV-infected children (P = 0.03). CONCLUSIONS: There is a high prevalence of HIV co-infection in children with TB. Progressive PTB and death are more common in HIV-infected children. Tuberculin skin testing is of limited use in screening for TB in HIV-infected children even when using a cut-point of > or = 5 mm. PMID- 10815740 TI - The impact of HIV infection on recurrence of tuberculosis in South African gold miners. AB - DESIGN AND OBJECTIVES: Potential risk factors for recurrence of tuberculosis (TB) were investigated in a retrospective cohort study of 305 human immunodeficiency virus (HIV) positive and 984 HIV-negative South African gold miners treated for TB with directly-observed, rifampicin-based regimens. Standard treatment changed from rifampicin, isoniazid and pyrazinamide (RHZ) to RHZ plus ethambutol (RHZE) during the study period. RESULTS: Recurrence occurred in 37 HIV-positive and 46 HIV-negative men. HIV infection was associated with a significantly higher recurrence rate (8.2 vs 2.2 per 100 person-years; multivariate-adjusted incidence rate ratio [IRR] 4.9, 95% confidence interval [CI] 3.0-8.1), as were post tuberculous scarring (multivariate-adjusted IRR 1.6 for one or two scarred lung zones, 4.0 for three or more zones; test for trend P < 0.001) and drug resistance (multivariate-adjusted IRR 2.7, 95%CI 1.01-7.4). The recurrence rate was significantly higher following treatment with RHZ than RHZE (multivariate adjusted IRR 2.1, 95%CI 1.1-4.0). The difference between regimens needs to be interpreted with caution, however, as allocation was not randomised. CONCLUSION: The high recurrence rate among HIV-positive men requires further investigation to distinguish relapse from re-infection as the predominant cause, leading to consideration of further intensification of the initial regimen or use of secondary prophylaxis. PMID- 10815741 TI - Comparison of IS1245 restriction fragment length polymorphism and pulsed-field gel electrophoresis for typing clinical isolates of Mycobacterium avium subsp avium. AB - SETTING: Little is still known about the epidemiology and pathogenesis of Mycobacterium avium subsp avium (MASA) infection. OBJECTIVE: Examination of the reproducibility and the stability over time of pulsed-field gel electrophoresis (PFGE) and IS1245 restriction fragment length polymorphism (IS1245-RFLP) techniques. The ability of these typing systems for differentiating clinical isolates of MASA was also assessed. DESIGN: Clinical isolates recovered from 63 patients (59 human immunodeficiency virus [HIV] positive and four HIV-negative) were studied by insertion sequence IS1245 and PFGE. For the study of in vivo and in vitro stability, strains collected over time from four patients and five strains chosen at random, respectively, were used. RESULTS: The stability of PFGE and IS1245-RFLP in vitro was excellent. PFGE was also stable in vivo, but IS1245 RFLP patterns showed some variation. The discriminatory power of IS1245-RFLP and PFGE was 0.995 and 0.989, respectively. The cluster analysis did not reveal differences between strains recovered from HIV-negative and HIV-positive patients or between patients with colonisation, local infection or disseminated disease. CONCLUSION: IS1245-RFLP and PFGE are useful tools for typing MASA strains. However, IS1245 variations in vivo may complicate the analysis of epidemiological relationships. PMID- 10815742 TI - Systemic but not intra-intestinal vaccination with BCG reduces the severity of tuberculosis infection in ferrets (Mustela furo). AB - SETTING: Ferrets are important wildlife vectors of bovine tuberculosis (Mycobacterium bovis) in New Zealand. By reducing the severity and/or incidence of tuberculosis (TB) in wild ferret populations, vaccination may limit disease transmission to livestock. OBJECTIVE: To investigate whether vaccination of ferrets with attenuated M. bovis BCG via systemic or intraintestinal routes can reduce the severity of TB resulting from oral M. bovis challenge. DESIGN: Groups of captive ferrets were vaccinated with live BCG via sub-cutaneous injection or intra-duodenal inoculation, twice, 4 weeks apart. Vaccinated and non-vaccinated (control) ferrets were subsequently challenged orally with virulent M. bovis to simulate the natural route of infection. Peripheral blood lymphocyte reactivity was longitudinally monitored, and the outcome of challenge was determined 20 weeks later by autopsy, histology and bacteriological culture. RESULT: Both vaccination routes induced tuberculin-specific lymphocyte reactivity; however, only the subcutaneous route was effective in reducing disease. Subcutaneous vaccinated ferrets had a lower severity of infection than non-vaccinated control animals, as indicated by significant reductions in viable bacterial burdens and prevention of gross lesions in mesenteric lymph nodes (the primary site of infection), and a lower incidence of bacterial translocation to thoracic lymph nodes. However, sub-cutaneous vaccination did not reduce the incidence of mesenteric lymph node infection. CONCLUSIONS: Systemic vaccination with BCG can reduce the severity of TB resulting from oral challenge with virulent M. bovis; however, delivery of viable BCG to the upper intestinal tract may not protect ferrets against TB. PMID- 10815743 TI - Detection of rifampicin resistance in Mycobacterium tuberculosis isolates from diverse countries by a commercial line probe assay as an initial indicator of multidrug resistance. AB - The line probe assay (LiPA), a rapid molecular method for detecting rifampicin resistance (RMPr) in Mycobacterium tuberculosis, correctly identified all 145 rifampicin-sensitive (RMPs) and 262 (98.5%) of 266 RMPr strains among 411 isolates collected from diverse countries. If used as a marker of multidrug resistant tuberculosis (MDR-TB), detection of RMPr by LiPA would have detected 236 of the 240 MDR strains in this study but would have wrongly suggested the presence of MDR in 26 RMP-monoresistant isolates (sensitivity 98.3%, specificity 84.8%). Hence, the reliability of using LiPA (or any other rapid RMPr-detection method) as a surrogate marker of MDR-TB largely depends on the prevalence of RMP monoresistance in the study population. This approach must therefore be validated in each local situation. PMID- 10815744 TI - How much isoniazid is needed for prevention of tuberculosis among immunocompetent adults? PMID- 10815745 TI - Tuberculosis and health sector reform: experience of integrating tuberculosis services into the district health system. PMID- 10815746 TI - [Monoarticular sterno-clavicular arthritic tuberculosis: a proposal and an observation]. PMID- 10815747 TI - Neurotoxic effects of three fractions isolated from Tityus serrulatus scorpion venom. AB - Scorpion venoms contain low molecular weight basic polypeptides, neurotoxins, that are the principal toxic agents. These toxins act on ion channels, promoting a derangement that may result in an abnormal release of neurotransmitters. In the present study we investigated some of the effects of the F, H and J fractions isolated from Tityus serrulatus scorpion venom on the central nervous system of rodents. The venom was partially purified by gel filtration chromatography. The neurotoxic effect of these fractions was studied on convulsive activity after intravenous injection, and on electrographic activity and neuronal integrity of rat hippocampus when injected directly into this brain area. The results showed that intravenous injection of the F and H fractions induced convulsions, and intrahippocampal injection caused electrographic seizures in rats and neuronal damage in specific hippocampal areas. Fraction J injected intravenously reduced the general activity of mice in the open field but induced no changes when injected into the brain. These results suggest that scorpion toxins are able to act directly on the central nervous system promoting behavioural, electrographic and histological modifications. PMID- 10815748 TI - Protective effect of ebselen on aflatoxin B1-induced cytotoxicity in primary rat hepatocytes. AB - Recent studies have shown that aflatoxin B1 enhances reactive oxygen species formation and causes oxidative damage, which may ultimately contribute to the cytotoxicity and carcinogenic effect of aflatoxin B1. Ebselen, 2-phenyl-1,2 benzoisoseleazol-3(H)-one, a synthetic seleno-organic compound has been shown to possess glutathione peroxidase-like activity and free radical scavenging ability. Thus present study was designed to investigate the protective effect of ebselen on aflatoxin B1-induced cytotoxicity in primary rat hepatocytes. Aflatoxin B1 induced cytotoxicity and lipid peroxidation were determined by lactate dehydrogenase leakage and malondialdehyde generation, respectively. Intracellular reactive oxygen species level was measured using the fluorescent probe 2',7' dichlorofluorescin diacetate, and the intracellular reduced glutathione concentration was determined with a fluorometric method. Ebselen was found to display a dose-dependent protective effect on lactate dehydrogenase leakage and malondialdehyde generation caused by aflatoxin B1 exposure. The results also demonstrate that ebselen efficiently inhibits the intracellular reactive oxygen species formation in aflatoxin B1-treated hepatocytes in a dose and time dependent manner. It was also noted that ebselen was able to increase the intracellular reduced glutathione concentration, both in the control and in aflatoxin B1-treated hepatocytes. The protection of ebselen against aflatoxin B1 cytotoxicity, however, was not affected by lowering the concentration of intracellular reduced glutathione. The overall data indicate that ebselen possesses a potent protective effect against aflatoxin B1-induced cytotoxicity, and the main mechanism involved in the protection may be its strong capability in inhibiting intracellular reactive oxygen species formation and preventing oxidative damage. PMID- 10815749 TI - Effect of nitric oxide synthase inhibitors on lipid peroxide formation in liver caused by endotoxin challenge. AB - This study investigated the effect of nitric oxide on lipid peroxide formation during endotoxaemia. Nitric oxide synthase inhibitors N(G)-monomethyl-L-arginine acetate (L-NMMA, 20 mg/kg, intravenously), N(G)-nitro-L-arginine-methyl ester (L NAME, 10 mg/kg, intravenously), and N(G)-nitro-L-arginine (L-NA, 10 mg/kg, intravenously), and a relatively selective inducible nitric oxide synthase inhibitor aminoguanidine (10 mg/kg, intravenously), did not protect against endotoxin-induced death of mice. Superoxide dismutase activity in liver 18 hr after administration of endotoxin (6 mg/kg, intraperitoneally) to L-arginine analogues (L-NMMA, L-NAME, L-NA)-treated mice was lower than in mice treated with endotoxin alone, whereas the administration of L-arginine analogues increased xanthine oxidase activity in the livers of endotoxin-injected mice compared with mice treated with endotoxin alone. In mice treated with L-arginine analogues and aminoguanidine, the levels of non-protein sulfhydryl and lipid peroxide in liver 18 hr after endotoxin injection did not show significant differences from mice treated with endotoxin alone. L-Arginine analogues and aminoguanidine had little effect on lipid peroxide formation in liver caused by endotoxin. Treatment with aminoguanidine (300 microM) significantly inhibited endotoxin-induced intracellular peroxide in J774A.1 cells, however, aminoguanidine did not affect endotoxin-induced cytotoxicity in J774A.1 cells. Our results clearly demonstrate that treatment with catalase (10 microg/ml), D-mannitol (10 mM), or superoxide dismutase (100 U/ml), has little or no effect on nitric oxide production by endotoxin (1 microg/ml)-activated J774A.1 cells. These findings suggest that nitric oxide is not crucial for lipid peroxide formation during endotoxaemia. Therefore, it is unlikely that nitric oxide plays a significant role in liver injury caused by free radical generation in endotoxaemia. PMID- 10815750 TI - Sensitivity to nitrogen mustard relates to the ability of processing DNA damage in Chinese hamster ovary cells. AB - The hallmark of the excision repair pathways is the removal of DNA adducts by excision of the damaged nucleotides. In the course of repair, transient DNA strand breaks occur, which can be measured by the Comet assay. We have investigated the processing of DNA damage, mediated by nitrogen mustard, in wild type AA8 Chinese hamster ovary cells, and in UV5, UV20 and UV41 DNA repair deficient cell lines. Whereas DNA repair could not be detected by unscheduled DNA synthesis at nitrogen mustard doses below 10 microM, processing of nitrogen mustard-mediated DNA damage was observed by the Comet assay at a 100-times lower concentration. Wild-type Chinese hamster ovary AA8 cells were able to process nitrogen mustard-mediated DNA damage within 4-24 hr depending on the dose of nitrogen mustard (0.1-10 microM). None of the repair-deficient cell lines was able to completely process the DNA damage induced by 10 microM nitrogen mustard. At nitrogen mustard doses that conferred 10% colony forming ability, the repair deficient cells had an altered processing of nitrogen mustard-mediated DNA damage: In the AA8, UV20, and UV41 cells, the amplitude of strand breaks peaked early (within 4 hr), the level of strand breaks in the nitrogen mustard exposed UV20 and UV41 cells did not return to the baseline of the unexposed reference culture, and the peak in strand breaks in the UV5 cell line occurred after 4 hr. Our results indicate that the single cell gel electrophoresis (Comet) assay is suitable for assessing repair capability of DNA alkylations. PMID- 10815751 TI - Bioavailability and pharmacokinetics of isradipine after oral and intravenous administration: half-life shorter than expected? AB - Isradipine is a calcium channel-blocking agent of the dihydropyridine type, used in the treatment of hypertension. A terminal half-life of 8-9 hr has been reported, in several pharmacokinetic studies after oral administration of isradipine. In a yet unpublished study a much shorter half-life was observed, and the present trial was therefore conducted in order to estimate the half-life after intravenous administration of isradipine. The bioavailability was estimated as well. In a randomised cross-over design ten healthy young volunteers were given either isradipine orally or an intravenous infusion. The two study periods were separated by at least 3 days. Blood samples for measurement of isradipine concentration were collected for 10-12 hr after administration and half-life and bioavailability were estimated. Mean terminal half-life after intravenous administration was calculated to be 2.8 hr, and the bioavailability to be 0.28. None of the 10 subjects suffered from side effects. In the present intravenous study the half-life of isradipine seems to be of much shorter than demonstrated in previous oral studies. PMID- 10815752 TI - Influence of ageing on vasomotor responses of human epicardial coronary arteries. AB - Vasomotor responses to various agonists were studied on isolated circular segments of human epicardial coronary arteries from three different age groups; 23-38 years, 40-58 years and 63-86 years. Noradrenaline had no or only weak contractile effect on coronary arteries from younger patients but induced contraction of all artery segments from older patients. The Emax value was significantly (P<0.0001) higher in arteries from the oldest group compared to each of the two younger age groups, whereas the potency was similar in all three groups. Linear regression analysis of noradrenaline-induced contraction in individual patients revealed a significantly positive age-correlation (correlation coefficient 0.67, P<0.0001). Contraction induced by endothelin-1 and relaxation induced by substance P, calcitonin gene-related peptide and vasoactive intestinal peptide on arteries precontracted with U46619 showed no significant differences in maximum responses and potencies between the three age groups, and no significant linear age-correlation. These data demonstrate a large variability in contractile responses to noradrenaline with contractions seen mostly in coronary arteries from older patients. It thus seems that sympathetic activation could contribute to coronary ischaemia in some patients. PMID- 10815753 TI - The effect of aminophylline on right heart function in young pigs after ligation of the right coronary artery. AB - An experimental model of right heart failure was developed to determine the effects of fluid loading and aminophylline on right heart function. We hypothesised that aminophylline would specifically improve right heart function through a decrease in pulmonary vascular resistance and, possibly, an increase in cardiac contractility. Right heart infarct was induced in ten experimental pigs and seven control pigs by ligating branches of the right coronary artery. The effect of fluid loading with a colloid solution and subsequent bolus doses of aminophylline on haemodynamics was observed. Fluid loading improved haemodynamics as expected. Aminophylline transiently improved cardiac index and pulmonary vascular resistance, but simultaneously caused an increase in heart rate and a decrease in stroke volume. Although aminophylline may reduce right heart afterload, it did not improve overall cardiac function in this experimental model of right heart infarction. PMID- 10815754 TI - Effects of troglitazone on insulin sensitivity in HIV-infected patients with protease inhibitor-associated diabetes mellitus. AB - Antiretroviral therapy (ART) is frequently associated with metabolic alterations, including insulin resistance and diabetes mellitus. In this pilot study, we evaluated the effect of the PPARgamma activator troglitazone on ART-associated insulin resistance in HIV-infected patients with ART-associated diabetes mellitus. Six patients with protease inhibitor (PI)-associated diabetes mellitus, lipodystrophy and dyslipidemia were treated with troglitazone 400 mg q.d. for 3 months. Previous oral antidiabetics were discontinued prior to the study. At baseline and after 3 months, insulin sensitivity (intravenous insulin tolerance test), body composition (multifrequence bioelectrical impedance analysis) and fat distribution (CT scan quantification) were assessed. Glycaemic control (fasting and postprandial blood glucose, fructosamine, glycosylated haemoglobin) and serum lipid status were determined monthly. In four of the six patients, there was a clear improvement in insulin sensitivity, resulting in a reversal of insulin resistance in two of these patients. Overall, there was an increase in lean body mass and a decrease in total body fat. The volume of visceral adipose tissue decreased whilst the volume of subcutaneous adipose tissue increased. Total cholesterol, LDL and HDL cholesterol increased, and total triglycerides and VLDL cholesterol decreased. No adverse effects such as hepatotoxicity were observed. Treatment with troglitazone 400 mg q.d. can ameliorate and in some cases even reverse ART-associated insulin resistance. Therefore, further studies including non-diabetic patients with ART-associated insulin resistance may be helpful in evaluating the long-term effects of thiazolidinediones on ART-associated insulin resistance and other metabolic complications, such as adipose maldistribution and dyslipidaemia. PMID- 10815755 TI - Efficiency of adenovirus-mediated gene transfer into hepatocytes by liver asanguineous perfusion method. AB - Efficient targeted gene delivery is essential for successful gene therapy. In this study, we examined the liver asanguineous perfusion method (LAP) for adenovirus-mediated gene transfer to the liver from the standpoints of efficiency, tissue-specificity and safety. The adenoviral vector containing the E. coli LacZ gene driven by the CAG promoter was delivered to the livers of rats by LAP. This method involves selective in situ perfusion, with the liver isolated by clamping of the afferent and efferent blood vessels to prevent adenoviral vector dissemination and genetic modification of nonhepatic organs. We demonstrated that gene transfer to the liver by LAP was not uniform, but more efficient than by intravenous (i.v.) or intraportal (i.p.) infusion, and caused no obvious liver damage or high mortality. As determined by specific histochemical staining and polymerase chain reaction, the amount of vector DNA transferred to the nonhepatic organs by LAP was significantly less than that transferred by the other two methods. Our data suggest that LAP is clearly superior to i.v. or i.p. infusion in terms of efficiency, specificity and safety of gene delivery to the liver. Further reduction in surgical risk is needed for the clinical application of gene therapy. PMID- 10815756 TI - Effects of galanin(1-16) on pancreatic secretion in anesthetized and conscious rats. AB - Galanin, a 29-amino acid peptide, has been demonstrated in pancreatic nerve endings and found to inhibit insulin release in the rat. However, the data available concerning its effects on exocrine pancreatic secretion are contradictory. The aim of the present study was to evaluate the effects of a synthetic porcine galanin sequence, Gal(1-16), on stimulated pancreatic secretion in hyperglycemic anesthetized and conscious rats. Male Wistar rats were anesthetized and surgically prepared with pancreatic and femoral vein catheters. In anesthetized animals, the pancreatic secretion was continuously stimulated with 150 ng cholecystokinin octapeptide (CCK-8)/kg body weight per 30 min, dissolved in saline or 10% glucose. Synthetic Gal(1-16) (0.3 or 1 nmol/kg per h) was infused over a 60-min period. In conscious rats, 1, 3, or 10 nmol Gal(1 16)/kg per h was administered in a continuous saline or 10% glucose infusion over a 30-min period. The pancreatic secretory volume and protein output were determined in 30-min samples in both models. In anesthetized rats, 0.3 nmol Gal(1 16)/kg per h did not modify pancreatic secretion during CCK-8 stimulation. However, both the pancreatic secretory volume and the protein output were significantly inhibited compared with the basal levels by 1 nmol Gal(1-16)/kg per h. The inhibitory effect of Gal(1-16) on pancreatic secretion was more marked with CCK-8/glucose (53.9%) than with CCK-8/saline stimulation (20.1%). In conscious rats, significant inhibitory effects of 1 nmol Gal(1-16)/kg per h in saline were observed (18%). During glucose infusion, a dose-dependent inhibition of 1, 3, and 10 nmol Gal(1-16)/kg per h on pancreatic secretory volume and protein output (35% inhibition at 1 nmol/kg per h) was observed. In conclusion, the inhibitory effect of Gal(1-16) on exogenous and endogenous CCK-stimulated pancreatic secretion was found to be more potent in the presence of glucose both in anesthetized and in conscious rats. These results may suggest an indirect (insulin-mediated) inhibitory effect of porcine Gal(1-16) on pancreatic secretion in the rat. PMID- 10815757 TI - Superoxide dismutase activity and the effects of NBQX and CPP on lipid peroxidation in experimental spinal cord injury. AB - The endogenous activity of the neuroprotective enzyme superoxide dismutase (SOD) and the amount of lipid peroxidation in the early phase of experimental spinal cord injury, together with the effects of N-methyl-D-aspartate (NMDA) antagonist CPP and non-NMDA antagonist NBQX on lipid peroxidation were evaluated. The clip compression model was used for the production of a standardized spinal cord trauma. SOD activity and malondialdehyde (MDA) levels--as an indicator of lipid peroxidation--were determined in the injured segment of the spinal cord 30 and 60 min after injury. SOD activity did not change in this period, whereas MDA levels at 30 and 60 min after trauma were significantly elevated. Intrathecal administration of CPP or NBQX 15 min after injury produced statistically significant reductions in MDA elevation 60 min after injury. NBQX was found to be more effective than CPP. These results demonstrated that intrathecal local application of excitatory amino acid receptor antagonists can protect the spinal cord from secondary damage caused by the generation of lipid peroxides in experimental spinal cord injury. PMID- 10815758 TI - Mechanism of antinephritic effect of proteinase inhibitors in experimental anti GBM glomerulopathy. AB - We have previously documented amelioration of rat autologous anti-GBM nephritis with the antiproteolytic drugs epsilon-aminocaproic acid (EACA) and aprotinin, given from the day of induction or later in the course of disease. In the present study we investigated potential mechanisms of this effect by assessing interactions of the drugs with proteinase-dependent generation of superoxide anion in glomeruli, and their influence on both GBM degradation in vitro and activity of glomerular proteolytic enzymes. Release of O2- by enzymatically disrupted glomeruli, isolated from nephritic control or EACA/aprotinin-treated rats, was measured with the ferricytochrome reduction method and its activity was correlated with proteinuria and glomerular cellularity at the early phase of the disease. The hydroxyproline release assay was used to quantitate degradation of rat GBM in vitro by leukocyte proteinases stimulated by phorbol myristate acetate (PMA), in the presence or absence of EACA and aprotinin. Finally, the activities of elastase, cathepsins B and L, and plasmin, together with collagenase-like activity, were assessed fluorimetrically in homogenates of glomeruli isolated from control and antiproteolytic-drug-treated nephritic rats. EACA and aprotinin notably inhibited production of superoxide by nephritic glomeruli (by 47% and 66%, respectively), and this effect was not significantly correlated with proteinuria or glomerular hypercellularity at the early stage of disease. On the other hand, generation of O2- by glomeruli of untreated nephritic rats was notably correlated with total glomerular cell counts and numbers of macrophages infiltrating glomeruli. PMA-stimulated neutrophils and macrophages caused degradation of isolated rat GBM in vitro, markedly attenuated in the presence of EACA (P<0.0005) and, to a lesser extent, by addition of aprotinin (P<0.01). The activity of elastase was significantly reduced in glomeruli of nephritic rats treated with EACA or aprotinin (both P<0.001), while activities of remaining proteinases were not appreciably affected. The beneficial influence of proteinase inhibitors on rat anti-GBM disease may be due, at least in part, to abrogation of superoxide generation in nephritic glomeruli. EACA and aprotinin also have potential to interfere with digestion of GBM, and both these effects may be related to suppression of glomerular elastase. PMID- 10815759 TI - Cell-cycle arrest and p53-independent induction of apoptosis in vitro by the new anticancer drugs 5-FdUrd-P-FdCydOct and dCydPam-P-FdUrd in DU-145 human prostate cancer cells. AB - PURPOSE: Current therapies have limited impact on the progression of metastatic hormone-refractory prostate cancer. Therefore, we investigated the utility of new heterodinucleoside phosphate dimers of 5-fluorodeoxyuridine (5-FdUrd) in p53 mutated and androgen-independent DU-145 human prostate tumour cells. METHODS: The effects of the dimers were assessed in vitro by a cell proliferation assay for cytotoxicity, flow cytometry for cell cycle distribution, confocal laser scanning microscopy for the detection of apoptotic bodies, poly(ADP-ribose) polymerase cleavage for caspase 3 activity and by a thymidylate synthetase assay. RESULTS: The new dimers N4-palmitoyl-2'-deoxycytidylyl-(3'-->5')-5-fluoro-2'-deoxyuridine (dCydPam-P-FdUrd) and 2'-deoxy-5-fluorouridylyl-(3'-->5')-2'-deoxy-5-fluoro-N4 octade cylcytidine (5-FdUrd-P-FdCydOct) caused marked cytotoxicity with IC50 values of 3-4 microM. 5-FdUrd-P-FdCydOct at 200 microM was capable of eradicating 100% of tumour cells whereas 10% of the cells were resistant to 5-FdUrd. Cytotoxicity was caused by a dramatic S-phase arrest, resulting in an increase of this cell population from 34% to 85% with 5-FdUrd-P-FdCydOct and to 81% with dCydPam-P-FdUrd. S-phase arrest was followed by apoptosis, as shown by 85% of the cells staining positive for Apo 2.7 antibody, a six- to eight-fold increased caspase 3 activity and DNA fragmentation. Thymidylate synthase activity was inhibited by 50% at 0.6-0.7 microM dimer concentration. The dimers were hydrolysed in vitro by phosphodiesterase I and human serum to the corresponding nucleosides and nucleoside monophosphates. CONCLUSIONS: The new dimers dCydPam-P FdUrd and 5-FdUrd-P-FdCydOct are effective prodrugs of 5-FdUrd and have potential value for the treatment of p53-mutated and hormone-independent human prostate carcinomas. PMID- 10815760 TI - Characterization of steroid hormone sensitivity in human breast cancers maintained ex vivo under organotypical culture conditions. AB - PURPOSE: The methodology we propose combines the immunohistochemical determination of the oestrogen and progesterone receptors (ER and PgR) with the characterization of the oestradiol- and progesterone-induced influence on cell proliferation in breast cancers in order to characterize their steroid hormone sensitivity at both the "static" and "dynamic" level. METHODS: ER and PgR have been immunohistochemically quantified by means of computer-assisted microscopy. Cell proliferation has been determined by means of tritiated thymidine autoradiography in tumour samples maintained in vitro as organotypic cultures. A series of 14 patients was investigated. RESULTS: Of the 14 breast cancers under study, one with an unequivocally "very ER-rich"/"very PgR-rich" immunohistochemical phenotype totally failed to exhibit any modification in its cell proliferation level after both oestradiol and progesterone stimulation. Two cases definitively associated with an "ER-poor"/"PgR-poor" immunohistochemical phenotype nevertheless responded noticeably to the dynamic stimulation of their cell proliferation by oestradiol and progesterone. While our series of cases covers 14 patients only, it suffices to demonstrate the limits of ER and PgR determination in characterizing steroid hormone sensitivity in breast cancer. DISCUSSION: The present work therefore presents an in vitro approach to test growth regulation of human breast cancer by steroid hormones. The clinical value of the present approach should be further determined by showing that steroid hormone-induced modifications in cell proliferation level are actually associated with clinical response. PMID- 10815761 TI - Effects of H-ras and v-sis overexpression on N-acetylglucosaminyltransferase V and metastasis-related phenotypes in human hepatocarcinoma cells. AB - Oncogenes and N-acetylglucosaminyltransferase (GnT-V) are both commonly associated with carcinogenesis and metastasis. In order to elucidate the relationship between oncogenes and GnT-V, two oncogenes, H-ras and v-sis/PDGF (platelet-derived growth factor), were selected, and the effects of their overexpression on GnT-V in 7721 human hepatocarcinoma cells were investigated. The results showed that the over expression of H-ras or v-sis/PDGF-B up-regulated the activities of GnT-V to various degrees in the transfected cells. In H-ras- and PDGF-B-overexpressing cells, the activity of GnT-V was up-regulated to double the normal value. The transient expression of v-sis, which produces a protein almost identical to PDGF-B, stimulated the GnT-V activity by 80.3%, and the effect was more pronounced (increased by 182.5%) in 7721 cells with stable expression of v-sis. The stimulating effect was entirely abolished by treatment with PDGF-B antibody. The staining of asparagine-linked glycans (N-glycans) in the H-ras- and v-sis-overexpressing 7721 cells was intensified when horseradish peroxidase-labeled leucoagglutinating phytohemogglutinin was used as a probe, indicating the increased content of beta1,6GlcNAc branching on the N-glycans. The enhancement of GnT-V mRNA expression was also observed in H-ras- and v-sis- overexpressing cells, indicating that H-ras and v-sis regulated GnT-V via the transcription of GnT-V mRNA and the synthesis of GnT-V protein. The cells overexpressing H-ras and v-sis displayed some changes in metastasis-related phenotypes, including acceleration of cell growth, decline of cell adhesion to fibronectin, and an increase of cell adhesion to laminin, as well as increased invasiveness through Matrigel. These results indicated that the alteration of cell adhesion and invasion induced by oncogenes is closely related to the up regulation of GnT-V activity and its product, beta1,6GlcNAc branching in N glycans on the cell surface. PMID- 10815762 TI - Bone morphogenetic protein 2 (BMP-2) induces sequential changes of Id gene expression in the breast cancer cell line MCF-7. AB - Bone morphogenetic proteins (BMPs) are involved in the development of various organs including the mammary gland. They are well-regulated and act in a time-, concentration- and cell-type-specific manner. We found that BMP-2 is expressed in primary breast tumor tissue samples and in breast cancer cell lines. Hybridization of labeled cDNA, obtained from the breast cancer cell line MCF-7, against the Atlas human cDNA expression array revealed differential gene expression depending on BMP-2 treatment. The most prominent changes were observed for the helix-loop-helix proteins Id-1, Id-2 and Id-3. Id-1 expression had increased severalfold after 4 h and was even higher after 24 h. Id-2 and Id-3 were more strongly induced after 4 h and showed no further significant change after 24 h. Analysis of cell-cycle distribution revealed a marked increase of the sub-G1 phase after 48 h in serum-deprived cells. In the presence of BMP-2 no change was observed over 48 h indicating that BMP-2 does not induce apoptosis. In addition, expression of caspase-3 was reduced in BMP-2-treated cells after 24 h. In summary, our results clearly indicate that BMP-2 is a susceptibility factor keeping the cells ready for the integration of various other signals for cell progression. PMID- 10815763 TI - Prognostic significance of DNA cytometry in thymoma. AB - PURPOSE: The aim of this work was to evaluate the prognostic significance of DNA image cytometry in thymoma. PATIENTS AND METHODS: Image cytometric studies with an automatic video-based analysis system (LEYTAS) were carried out on 47 archival specimens from 36 patients with thymomas who underwent operation at a single institution from 1954 to 1992. The significance of aneuploidy DNA-content (5c exceeding events), and nuclear size on stage and survival were evaluated. The median follow-up was 52.7 (6-164) months. RESULTS: Masaoka's stage was predictive of aneuploidy (P < 0.01) and disease-free survival (P < 0.015). In stage I 18% of the tumors were aneuploid, in stage II 78%, in stage III 85% and in stage IV 100%. The occurrence of 5c-exceeding events was associated with both decreased disease-free survival (P < 0.01) and overall survival (P = 0.013). Nuclear size was not significantly correlated to stage. Under multivariate analysis, aneuploidy and DNA content failed to attain independent significance for stage, performance status, and histology. CONCLUSION: DNA image cytometry may provide additional information about the prognosis of resected thymoma. PMID- 10815765 TI - Alterations in the estrogen receptor alpha mRNA in the breast tumors of African American women. AB - Several recent reports have shown that the mortality rate with breast cancer is about three times higher in African American women than in other populations. In addition, the available data also indicate that the tumors are very aggressive and poorly differentiated with a very low frequency of hormone receptors. To gain an insight into the factors that may be responsible for their aggressive tumors, we investigated the transcript profiles of the estrogen receptor (ER), the most important prognostic factor in breast cancer, in the tumors derived from African American women. We analyzed 24 immunohistochemically ER+ and 6 ER- malignant tumors for ER mRNA by reverse transcription polymerase chain reaction using a number of primer pairs. For comparative purposes, 20 ER- malignant tumor issues derived from Caucasian patients were also included. Our results showed that only 15 of the ER+ tumors from African American women patients had full-length wild type receptor transcripts and the others exhibited alterations/truncations in exon 8. We also found that the majority of tumors that had alterations/truncations in exon 8 did not express the naturally occurring, more abundant exon 7 deletion transcript. Most of the tumors expressed exon 2, exons 2 3, and exon 5 deletion variant transcripts. Unexpectedly, 2 of the 6 immunohistochemically ER- tumors showed full-length wild-type receptor mRNA but none of the variant transcripts. PMID- 10815764 TI - Computer-assisted image analysis can aid the prognostication of advanced-stage neuroblastomas. AB - PURPOSE: The aim of this work was to detect nuclear parameters related to the prognosis of patients with stage III, IV or DS neuroblastomas. METHODS: Histological sections of 25 operation specimens obtained from children with advanced-stage neuroblastomas were subjected to computer-assisted image analysis. Statistical relationships between nuclear descriptors of the tumor cells and patients' clinical outcome were determined. RESULTS: The coefficient of variability of the mean nuclear area the mean nuclear elongation factor, and the mean nuclear averaged Feret diameter of the neuroblastoma cells were ascertained to be discriminators separating high-grade from low-grade tumors. CONCLUSIONS: The histomorphometrically gauged nuclear parameters may help oncologists to assess the prognosis of patients with advanced-stage neuroblastoma. PMID- 10815766 TI - Experience with prophylactic oral ciprofloxacin in gynecological cancer patients developing severe chemotherapy-induced neutropenia. AB - The development of significant neutropenia is a relatively frequent complication of cytotoxic chemotherapy of malignant disease. In an effort to develop a cost effective management strategy to prevent serious infectious events associated with severe chemotherapy-induced neutropenia, patients treated in the Gynecological Oncology Program of the Cleveland Clinic Taussig Cancer Center have been administered ciprofloxacin, a potent broad-spectrum antibiotic, 500 mg orally twice a day, beginning at the time of documentation of grade 4 neutropenia. The antibiotic is continued until granulocyte recovery. A total of 44 patients (57 treatment courses) have been treated in this manner. There have been no complications of therapy and no episodes of subsequent infections due to ciprofloxacin-resistant organisms. Two patients required hospital admission following the development of significant fever, despite the use of ciprofloxacin. While these results are encouraging and suggest that the use of prophylactic ciprofloxacin is a highly cost-effective management approach in this setting, randomized controlled trials are necessary to define the ultimate benefit of this clinical strategy. PMID- 10815767 TI - Conference on the occasion of the 20th anniversary of the Psychosocial Care Unit at the Surgical University Hospital Heidelberg. Looking back into the future- experiences, viewpoints and prospects in psychosocial oncology. 8-9 October 1999, Heidelberg, Germany. PMID- 10815768 TI - Cyclophosphamides as hypoxia-activated diffusible cytotoxins: a theoretical study. AB - Cyclophosphamides have been in clinical use as anti-cancer drugs for a long time and much research has been directed towards reducing their side effects. Here we have performed a theoretical investigation into the possibility of designing bioreductive analogues of cyclophosphamides. Our calculations have employed semiempirical molecular orbital AM1-SM2 and PM3-SM3 calculations, as implemented in MOPAC 93, which include a modified Born method for the treatment of solvation. We have investigated the effect of bioreductive activation on the beta elimination reaction that is central to the activation of cyclophosphamides. The approach was tested on two known bioreductive agents, including CB1954, and gave results in agreement with experiment. Non-local density functional calculations on CB1954 and its metabolites, including the radical anion, were in agreement with the semiempirical calculations. The calculations have identified a number of potentially novel bioreductive cyclophosphamides. In particular, our calculations identified compounds in which the initial one-electron reduction was not activating. Such compounds are likely to be more effective bioreductive agents, as the beta-elimination will not compete under oxic conditions with the important re-oxidation required for the protection of oxic tissue. PMID- 10815769 TI - Structural analysis of the KGD sequence loop of barbourin, an alphaIIbbeta3 specific disintegrin. AB - Disintegrins constitute a class of small proteins that inhibit platelet aggregation by binding to the fibrinogen receptor, also referred to as integrin alphaIIbbeta3. Contrarily to other disintegrins that bind to a series of integrins via their Arg-Gly-Asp domain, the recognition site of barbourin contains a Lys-Gly-Asp sequence that ensures its specificity towards alphaIIbbeta3. In this article, a three-dimensional model of barbourin is proposed using homology modeling and large-scale molecular dynamics simulations. The conformations of the Lys-Gly-Asp sequence of barbourin are analyzed and compared to those of peptidomimetics that exhibit similar specificity towards alphaIIbbeta3. The tryptophan residue following the Lys-Gly-Asp sequence of the binding domain is shown to play a crucial role in the biological activity and the specificity of barbourin. Our results suggest that this disintegrin anchors to the binding pocket of the gamma-chain of fibrinogen rather than to those of the Arg-Gly-Asp sequence. PMID- 10815770 TI - Mixed QM/MM molecular electrostatic potentials. AB - A new method is presented for the calculation of the Molecular Electrostatic Potential (MEP) in large systems. Based on the mixed Quantum Mechanics/Molecular Mechanics (QM/MM) approach, the method assumes both a quantum and classical description for the molecule, and the calculation of the MEP in the space surrounding the molecule is made using this dual treatment. The MEP at points close to the molecule is computed using a full QM formalism, while a pure classical evaluation of the MEP is used for points located at large distances from the molecule. The algorithm allows the user to select the desired level of accuracy in the MEP, so that the definition of the regions where the MEP is computed at the classical or QM levels is adjusted automatically. The potential use of this QM/MM MEP in molecular modeling studies is discussed. PMID- 10815771 TI - 3D-QSAR methods on the basis of ligand-receptor complexes. Application of COMBINE and GRID/GOLPE methodologies to a series of CYP1A2 ligands. AB - Many heterocyclic amines (HCA) present in cooked food exert a genotoxic activity when they are metabolised (N-oxidated) by the human cytochrome P450 1A2 (CYP1A2h). In order to rationalize the observed differences in activity of this enzyme on a series of 12 HCA, 3D-QSAR methods were applied on the basis of models of HCA-CYP1A2h complexes. The CYP1A2h enzyme model has been previously reported and was built by homology modeling based on cytochrome P450 BM3. The complexes were automatically generated applying the AUTODOCK software and refined using AMBER. A COMBINE analysis on the complexes identified the most important enzyme ligand interactions that account for the differences in activity within the series. A GRID/GOLPE analysis was then performed on just the ligands, in the conformations and orientations found in the modeled complexes. The results from both methods were concordant and confirmed the advantages of incorporating structural information from series of ligand-receptor complexes into 3D-QSAR methodologies. PMID- 10815772 TI - Research on anti-HIV-1 agents. Investigation on the CD4-Suradista binding mode through docking experiments. AB - Sulfonated distamycin (Suradista) derivatives exhibit anti-HIV-1 activity by inhibiting the binding of the viral envelope glycoprotein gp120 to its receptor (CD4). With the aim to propose a possible binding mode between Suradistas and the CD4 macromolecule, molecular docking experiments, followed by energy minimization of the complexes thus obtained, were performed. Computational results show that ligand binding at the CD4 surface involves two or three positively charged regions of the macromolecule, in agreement with the results of X-ray crystallographic analysis of a ternary complex (CD4/gp120/neutralizing antibody) recently reported in the literature. Our findings account well for the structure activity relationship found for Suradista compounds. PMID- 10815773 TI - Conformational analysis of a farnesyltransferase peptide inhibitor, CVIM. AB - The conformational states of the peptide Cys-Val-Ile-Met (or CVIM) were computed and characterized. CVIM inhibits farnesylation of the Ras oncogene product, p21ras, at the cysteine residue of the C-terminal segment. CVIM is active in an extended conformation. A similar peptide (KTKCVFM) appears to bind the enzyme in the Type I bend conformation. In the present study, the conformations of CVIM were computed in an aqueous environment with the peptide in the zwitterionic state. Solvation free energy based on solvent accessible surface area and a distance dependent dielectric were used in the calculations. Final conformations of multiple independent Monte Carlo simulated annealing (MCSA) conformational searches were used as starting points for Metropolis Monte Carlo (MMC) runs. Conformations saved at intervals during MMC runs were analyzed. Conformers were separated by interactive clustering in dihedral angle coordinates. The four lowest energy conformers corresponding to a Type I bend, extended, AB-bend, and BA-bend were within 0.3 kcal/mol of each other, and dominant in terms of population. The Type I bend and extended conformers were supported by the binding studies. The extended conformer was the most populated. In the AB-bend conformer, 'A' indicates the alpha-helix conformation of Val, and 'B' indicates the beta strand conformation of Ile. The AB- and BA-bend conformations differed from the extended conformation in the value of Val psi and Ile psi, respectively, and from the Type I bend conformation in the value of Ile psi and Val psi, respectively. The four lowest energy conformers were characterized in terms of energy, density of low energy conformations (or entropy), structure, side chain rotamer fraction population, and interatomic distances. PMID- 10815774 TI - Fast prediction and visualization of protein binding pockets with PASS. AB - PASS (Putative Active Sites with Spheres) is a simple computational tool that uses geometry to characterize regions of buried volume in proteins and to identify positions likely to represent binding sites based upon the size, shape, and burial extent of these volumes. Its utility as a predictive tool for binding site identification is tested by predicting known binding sites of proteins in the PDB using both complexed macromolecules and their corresponding apoprotein structures. The results indicate that PASS can serve as a front-end to fast docking. The main utility of PASS lies in the fact that it can analyze a moderate size protein (approximately 30 kDa) in under 20 s, which makes it suitable for interactive molecular modeling, protein database analysis, and aggressive virtual screening efforts. As a modeling tool, PASS (i) rapidly identifies favorable regions of the protein surface, (ii) simplifies visualization of residues modulating binding in these regions, and (iii) provides a means of directly visualizing buried volume, which is often inferred indirectly from curvature in a surface representation. PASS produces output in the form of standard PDB files, which are suitable for any modeling package, and provides script files to simplify visualization in Cerius2, InsightII, MOE, Quanta, RasMol, and Sybyl. PASS is freely available to all. PMID- 10815775 TI - Metaphoric structuring: understanding time through spatial metaphors. AB - The present paper evaluates the claim that abstract conceptual domains are structured through metaphorical mappings from domains grounded directly in experience. In particular, the paper asks whether the abstract domain of time gets its relational structure from the more concrete domain of space. Relational similarities between space and time are outlined along with several explanations of how these similarities may have arisen. Three experiments designed to distinguish between these explanations are described. The results indicate that (1) the domains of space and time do share conceptual structure, (2) spatial relational information is just as useful for thinking about time as temporal information, and (3) with frequent use, mappings between space and time come to be stored in the domain of time and so thinking about time does not necessarily require access to spatial schemas. These findings provide some of the first empirical evidence for Metaphoric Structuring. It appears that abstract domains such as time are indeed shaped by metaphorical mappings from more concrete and experiential domains such as space. PMID- 10815776 TI - Why we are so good at catching cheaters. AB - Data that appear to exhibit a 'cheater detection' effect on performance in the Wason Selection task are widely interpreted as implying that deontic reasoning is effected by a domain specific, cognitive module. The 'cheater detection module' is said to offer a clear example of an effect of evolutionary selection on human cognitive architecture. This interpretation depends critically on assuming that deontic conditionals and their indicative controls are identical in structure; hence that the asymmetries in S's performance must be effects of content variables. I argue that this assumption is untrue and that structural features of deontic conditionals predict the 'cheater detection' data without assumptions about either the architecture or the history of cognition. According to this analysis, the putative cheater detection effect on the Wason task is actually a materials artifact. PMID- 10815777 TI - Lexical word formation in children with grammatical SLI: a grammar-specific versus an input-processing deficit? AB - An ongoing controversy is whether an input-processing deficit or a grammar specific deficit causes specific language impairment (SLI) in children. Previous studies have focussed on SLI childrens' omission of inflectional morphemes or impaired performance on language tasks, but such data can be accounted for by either theory. To distinguish between these theories we study compound formation in a subgroup of SLI children with 'grammatical (G)-SLI'. An input-processing account (e.g. Leonard, L. (1998). Children with specific language impairment. Cambridge, MA: MIT Press), in which perception and production of inflections requires extra processing resources, would predict that G-SLI children will omit the regular plural -s in compounds (e.g. rat-eater). A grammar-specific deficit account (e.g. Ullman, M. & Gopnik, M. (1994) The production of inflectional morphology in hereditary specific language impairment. The McGill Working Papers in Linguistics, 10, 81-118; van der Lely, H. K. J. & Ullman, M. (1996). The computation and representation of past-tense morphology in normally developing and specifically language impaired children. In A. Stringfellow, D. Cahana Amitay, E. Hughes & A. Zukowski, Proceedings of the 20th Annual Boston University Conference on Language Development (pp. 816-827). Somerville, MA: Cascadilla Press), in which G-SLI children are impaired in regular inflectional morphology, would predict that G-SLI children will produce regular plural -s forms inside compounds (e.g. *rats-eater). We compared the responses of 16 G-SLI subjects (aged 10 years 4 months to 18 years) with those of 36 normally developing control children (24 matched on language ability and 12 matched on age and cognitive ability). All the groups produced irregular plural nouns in compounds (mice eater). The normally developing children and teenagers rarely, if ever. produced regular plural nouns inside compounds (*rats-eater), whereas the G-SLI subjects did so often. This pattern of results conflicts with the predictions ofthe input processing deficit account. The findings support the grammar-specific deficit hypothesis. The data rovide further evidence that specialized grammatical abilities may be differentially impaired within the language system. PMID- 10815778 TI - Verbal and visual causal arguments. AB - The present paper analyzes how verbalizations and visualizations can be used to justify and dispute causal claims. The analysis is based on a taxonomy of 27 causal arguments as they appear in ordinary language. It is shown how arguments from spatio-temporal contiguity, covariation, counterfactual necessity, and causal mechanisms, to name only a few, are visualized in persuasive uses of tables, graphs, time series, causal diagrams, drawings, maps, animations, photos, movies, and simulations. The discussion centers on how these visual media limit the argumentative moves of justifying, disputing, and qualifying claims; how they constrain the representation of observational, explanatory, and abstract knowledge in the premises of causal arguments; and how they support and externalize argument-specific inferences, namely generalizations, comparisons, mental simulations, and causal explanations. PMID- 10815779 TI - Graphemes are perceptual reading units. AB - Graphemes are commonly defined as the written representation of phonemes. For example, the word 'BREAD' is composed of the four phonemes /b/, /r/, /e/ and /d/, and consequently, of the four graphemes 'B', 'R', 'EA', and 'D'. Graphemes can thus be considered the minimal 'functional bridges' in the mapping between orthography and phonology. In the present study, we investigated the hypothesis that graphemes are processed as perceptual units by the reading system. If the reading system processes graphemes as units, then detecting a letter in a word should be harder when this letter is embedded in a multi-letter grapheme than when it corresponds to a single-letter grapheme. In Experiment 1A, done in English, participants were slower to detect a target letter in a word when the target letter was embedded in multi-letter grapheme (i.e. 'A' in 'BEACH') than when it corresponded to a single-letter grapheme (i.e. 'A' in 'PLACE'). In Experiment 1B, this effect was replicated in French. In Experiment 2, done in English, this grapheme effect remained when phonemic similarity between the target letter alone and the target letter inside the word was controlled. Together, the results are consistent with the assumption that graphemes are processed as perceptual reading units in alphabetic writing systems such as English or French. PMID- 10815780 TI - Relationship between hematopoietic growth factors levels and hematological parameters in Argentine hemorrhagic fever. AB - Argentine hemorrhagic fever (AHF) is a viral disease caused by Junin virus and characterized by hematologic and neurological involvement. The main hematologic features are leukopenia, thrombocytopenia, and bone marrow hypoplasia. Hematopoietic growth factors serum levels were measured by ELISA technique in forty-eight patients with confirmed diagnosis of AHF. Patients were classified according to the clinical picture in 15 severe (SCF), 17 moderate (MoCF), and 16 mild (MiCF) cases. Erythropoietin levels were decreased in 28 of 45 patients and raised in 4 SCF patients. Twenty-four of 38 patients had high G-CSF levels at admittance in accordance with clinical picture severity, while IL-3, GM-CSF, and TGF-beta were normal in most cases. A direct correlation was found between G-CSF and TNF-alpha levels. Thrombopoietin levels were found to be raised in 19 of 21 patients. In conclusion, the low levels of Epo may contribute to the severe bone marrow erythroblastopenia described in AHF patients, while G-CSF seems to be a marker of illness severity. PMID- 10815781 TI - Beta-thalassaemia in Cubans: novel allele increases the genetic diversity at the HBB locus in the Caribbean. AB - In order to establish the molecular basis of beta-thalassaemia in Cubans, a total of 75 unrelated individuals, with beta-thalassaemia major (7), Hb S-beta thalassaemia (28), Hb C-beta-thalassaemia (1), and beta-thalassaemia trait (39) yielding 82 beta-thalassaemia alleles, were analyzed. Seventeen different point mutations were identified accounting for 93% of the beta-thalassaemia alleles studied, revealing a high genetic heterogeneity at the HBB locus in this population. The more prevalent mutations, namely, CD 39 (C --> T) (30.5%), -29 (A --> G) (13.4%), IVS-I-110 (G --> A) (8.5%), and IVS-II-1 (G --> A) (8.5%), reflect the Mediterranean and African predominant ancestry of the extant Cuban population. We also report the identification of a novel allele, IVS-I-108 (T --> C), that possibly activates a cryptic branch site, in a beta-thalassaemia carrier with no other molecular defect within the beta-globin gene and its proximal promoter. This study shows that prenatal diagnosis of beta-thalassaemia should be feasible in about 60% of at-risk pregnancies by direct detection of selected point mutations. However, due to the wide spectrum of mutations, and in order to offer fully informative prenatal diagnosis to more than 87% of at-risk couples, the screening for beta-thalassaemia mutations in Cubans should be performed by using a general point mutation detection method, such as DGGE (denaturing gradient gel electrophoresis). PMID- 10815782 TI - Sequential nitric oxide measurements during the emergency department treatment of acute vasoocclusive sickle cell crisis. AB - This prospective study was designed to examine the relationship between serial serum nitric oxide (NO) levels and pain during the emergency department (ED) treatment of acute vasoocclusive sickle cell crisis (SCC). 102 patient visits, age > or =18 years of age, presenting to the ED with uncomplicated, typical SCC pain had serum NO levels obtained at 2-hr intervals during treatment of pain and were measured using an NO-specific chemiluminesence technique. Pain was measured prior to each NO measurement using a 10 cm visual analog scale (VAS), and subjects were divided into a persistent pain group and an improved pain group. Patients with persistent pain had significantly low initial NO levels (11.51 microM +/- 2.8, P < 0.05) while those with pain improvement had higher initial NO levels (18.1 microM +/- 3.08, P < 0.05). There was no significant correlation between changes in NO and changes in pain scores. These results suggest that the initial NO level may serve as a marker for the severity of tissue ischemia. Sequential NO levels do not appear useful in predicting the course of SCC. PMID- 10815783 TI - Effects of hypoxia on granulocytic-monocytic progenitors in rats. Role of bone marrow stroma. AB - Hemorrhagic shock leads to hypoxia and is associated with bone marrow (BM) failure. Hemorrhagic shock is also a predisposing factor in immune dysregulation. Since the BM is the major organ of immune cells in the adult, its failure following hemorrhagic shock may explain the increased susceptibility to infection. The in vitro evidence indicates that hypoxia mediates altered functions in BM stroma. Since similar hematopoietic alterations are reported in hypoxia and hemorrhagic shock, hypoxia alone could be a representative model to study BM responses during hemorrhagic shock. In this study, we use an animal model to dissect the hematopoietic effects of hypoxia. We subjected rats to hypoxia, and at days 1 and 5 post-hypoxia we determined the numbers of granulocytic-monocytic progenitors (CFU-GM) in the BM. We found significant increase (P < 0.05) in CFU-GM at day 1 and a downward trend by day 5. Enhanced BM cellularity could not explain the increase in CFU-GM by day 1. BM stromal cells mediated most of the stimulatory effects by hypoxia. CFU-GM was inversely proportional to bioactive TGF-beta and directly proportional to IL-1. Compared to normoxic rats, IL-6 production was suppressed in BM cells from hypoxic rats. The results show that hypoxia alone initiate a stimulatory response in CFU-GM progenitors. These effects are at least partially mediated through the BM stroma. In the absence of a second insult, CFU-GM reverts to baseline. The data also suggest that hypoxia mediates complex responses that include cytokine production. These results add to the current understanding of hematopoietic responses by hypoxia and adds to the mechanisms of immune dysfunctions following hemorrhagic shock. PMID- 10815784 TI - Cost-effectiveness of hydroxyurea in sickle cell anemia. Investigators of the Multicenter Study of Hydroxyurea in Sickle Cell Anemia. AB - The Multicenter Study of Hydroxyurea in Sickle Cell Anemia (MSH) demonstrated the efficacy of hydroxyurea in reducing the rate of painful crises compared to placebo. We used resource utilization data collected in the MSH to determine the cost-effectiveness of hydroxyurea. The MSH was a randomized, placebo-controlled double-blind clinical trial involving 299 patients at 21 sites. The primary outcome, visit to a medical facility, was one of the criteria to define occurrence of painful crisis. Cost estimates were applied to all outpatient and emergency department visits and inpatient hospital stays that were classified as a crisis. Other resources for which cost estimates were applied included hospitalization for chest syndrome, analgesics received, hydroxyurea dosing, laboratory testing, and clinic visits for management of patient care. Annualized differential costs were calculated between hydroxyurea- and placebo-receiving patients. Hospitalization for painful crisis accounted for the majority of costs in both arms of the study, with an annual mean of $12,160 (95% CI: $9,440, $14,880) for hydroxyurea and $17,290 (95% CI: $13,010, $21,570) for placebo. The difference in means was $5,130 (95% CI: $60, $10,200; P = 0.048). Chest syndrome was the next largest cost with a mean difference of $830 (95% CI: $-340, $2,000; P = 0.16). The hydroxyurea arm was also associated with lower costs for emergency department visits, transfusion, and use of opiate analgesics. In total, the annual average cost per patient receiving hydroxyurea was $16,810 (95% CI: $13,350, $20,270) and the annual average costs per patient receiving placebo was $22,020 (95% CI: $17,340, $26,710). The difference in means was $5,210 (95% CI: $ 610, $11,030; P = 0.21). The cost of hydroxyurea with the more intensive monitoring required when using this drug appears to be more than offset by decreased costs for medical care of painful crisis and analgesic use. Although the total cost difference was not significant statistically, these results suggest that hydroxyurea therapy is cost-effective compared to placebo in the management of adult patients with sickle cell anemia. If hydroxyurea can prevent development of chronic organ damage, long-term savings may be even greater. PMID- 10815785 TI - The Japanese multicenter open randomized trial of ursodeoxycholic acid prophylaxis for hepatic veno-occlusive disease after stem cell transplantation. AB - Hepatic veno-occlusive disease (VOD) is a common transplant-related complication of stem cell transplantation. There is no safe and proven therapy for established VOD, and attempts have focused on its prevention. Limited studies have suggested that prophylactic use of ursodeoxycholic acid (UDCA) reduced the incidence of VOD. To confirm the preventive effect of UDCA on VOD, we conducted a prospective, unblinded randomized, multicenter study of UDCA involving 132 patients who underwent stem cell transplantation for a variety of disorders. Sixty-seven patients were assigned to the UDCA-treated group, and 65 patients were assigned to the control group. The clinical characteristics of the two groups were similar with respect to primary diagnosis, age, sex, and baseline organ function. The preparative regimen and GVHD prophylaxis did not differ significantly between the two groups. UDCA was highly effective in preventing VOD, which occurred in only 3.0% in the UDCA-treated group, as opposed to 18.5% in the control group (P = 0.0043). There were no adverse effects attributable to UDCA. The initial promising report of a prophylactic effect of UDCA on VOD after stem cell transplantation was confirmed in this prospective study. PMID- 10815786 TI - Genetic epidemiology of HbS in Oman: multicentric origin for the betaS gene. AB - On the basis of a sample of 117 chromosomes, we have demonstrated the multicentric origin of the sickle mutation in Northern Oman. Three major haplotypes coexist: 52.1% Benin (typical and atypicals), 26.7% Arab-India, and 21.4% Bantu. These haplotypes are not autochthonous to Oman but originated elsewhere and arrived in Oman by gene flow. The distribution of haplotypes is in excellent agreement with the historical record, which establishes clear ancient contacts between Oman and sub-Sahara west Africa and explains the presence of the Benin haplotype; contacts with Iraq, Iran, present-day Pakistan, and India explain the presence of the Arab-India haplotype. More recent contacts with East Africa (Zanzibar/Mombasa) explain the presence of the Bantu haplotype. The pattern of the Arab-India haplotype in the populations of the Arabian peninsula reinforces the hypothesis that this particular mutation originated in the Harappa culture or in a nearby population and in addition reveals that the Sassanian Empire might have been the vehicle by which this Indo-European sickle mutation migrated (gene flow) to the present-day Arabian peninsula, including Oman. PMID- 10815787 TI - Maximum urine concentrating ability in children with Hb SC disease: effects of hydroxyurea. AB - Studies in adults with Hb SC disease suggested that hydroxyurea reduced hemolysis and increased red cell hydration. Because increased hydration should diminish the polymerization tendency of Hb S we hypothesized that hydroxyurea might repair the urine concentration defect of HbSC disease. Eight Hb SC disease patients, aged 10 to 17 years, were given hydroxyurea daily. Maximal urine concentrating ability following overnight fasting and after subcutaneous arginine vasopressin (dDAVP), blood counts, and cell volumes were observed for 12-15 months. All patients had impaired urine concentrating ability prior to hydroxyurea treatment and failed to increase their ability to concentrate urine following treatment (maximum urine concentration after an overnight fast and dDAVP, 520-530 mOsm). Mean corpuscular volume (MCV) and reticulocyte MCV increased after administration of hydroxyurea, and the reticulocyte count and ratio of red cell hemoglobin to reticulocyte hemoglobin fell but there was little change in PCV. Hb F increased substantially in 2 patients but showed little change in the remaining patients. There was no evidence that hydroxyurea was associated with increased urine concentrating ability in children with Hb SC disease. These results may reflect irreversible renal medullary damage prior to beginning treatment or insufficient intensity or duration of treatment. PMID- 10815788 TI - Familial myelodysplastic syndrome with early age of onset. AB - A family is described in which three members, the propositus, his brother, and son, developed a myelodysplastic syndrome (MDS) at the ages of 52, 35, and 25, respectively. A fourth member, the paternal uncle of the propositus, was diagnosed with chronic lymphocytic leukemia. Two of the three affected Individuals had megaloblastoid marrows with recognizable bone marrow cytogenetic abnormalities and progressive, nonleukemic bone marrow failure. The propositus was unresponsive to G-CSF and eventually died of sepsis. The second affected family member died of bone marrow transplant complications. The third affected family member underwent bone marrow transplantation and is showing signs of graft survival despite minor complications. The affected members of this pedigree appear to represent a continuum in severity of disease and, therefore, pathogenesis. The pattern of inheritance and clinical progression of the disease suggest a genetic defect which may predispose individuals to the development of MDS. PMID- 10815789 TI - Management of a patient with a mechanical aortic valve and antibodies to both thrombin and factor V after repeat exposure to fibrin sealant. AB - We describe a patient who developed a markedly prolonged PT, PTT, and thrombin time 13 days after repeat exposure to fibrin sealant during coronary artery bypass grafting and aortic valve replacement. Evaluation revealed an inhibitor to bovine thrombin that cross-reacted with human thrombin. In addition an inhibitor to human coagulation factor V was identified. Despite coagulation abnormalities there was no evidence of bleeding. Nevertheless, effective anticoagulation was required to minimize the thrombotic complications associated with the patient's prosthetic valve. We elected to take a conservative approach and not utilize pharmacologic anticoagulation until there was diminution in the effect of the acquired inhibitors. We report on our patient's course and review the available literature addressing the management of patients demonstrating inhibitors to blood coagulation factors after repeat exposure to fibrin sealants. PMID- 10815790 TI - HTLV-1 unrelated adult T-cell leukemia/lymphoma with unique phenotype and karyotype. AB - We describe a unique case of adult T-cell leukemia/lymphoma (ATL). The patient had typical clinicohematological features as ATL, but showed a lack of antibody to human T-cell leukemia virus type-1 (HTLV-1) and was negative for HTLV-1 proviral DNA in the peripheral mononuclear cells by means of polymerase chain reaction. The phenotype of tumor cells revealed CD7+, CD5+, CD2+, CD3+, WT31-, TcR delta 1-, CD4-, CD8-, CD25-, and the karyotype showed a 5q-, t(12;18). HTLV-1 unrelated ATL is very rare, and the karyotype as in our case has not been reported previously. PMID- 10815791 TI - Drug-induced hemolysis: cefotetan-dependent hemolytic anemia mimicking an acute intravascular immune transfusion reaction. AB - Numerous cases of drug-induced hemolytic anemia have been described in patients treated with penicillin or cephalosporin. Second and third generation cephalosporins are more commonly implicated in hemolytic reactions than first generation cephalosporins. We report a case of severe cefotetan-induced hemolytic anemia in a previously healthy 46-year-old woman undergoing an elective hysterectomy. The patient received 2 g of intravenous cefotetan intraoperatively and subsequently at 12 and 24 h post-operatively. She complained of diarrhea and fever on the third post-operative day and was seen in her gynecologist's office on the fifth post-operative day (hemoglobin = 10.5 g/dL). On the seventh post operative day, she complained of fever and soreness around the suprapubic catheter site and was given a prescription for 500 mg oral cephalexin four times a day. The next day she was seen in the gynecologist's office and reported feeling better. Ten days after the operation her fatigue worsened and her hemoglobin was 4.8 g/dL. She was transfused with 3 units of packed red blood cells (PRBC) and was given 1 g of cefotetan intravenously. During the transfusion of the second unit of PRBC nursing staff observed gross hemoglobinuria and she subsequently developed acute renal failure. Laboratory chemistry parameters were consistent with severe acute hemolysis. The patient's direct antiglobulin test was reactive and her serum reacted with cefotetan-coated red blood cells (RBCs) and serum plus soluble cefotetan reacted with untreated RBCs. The titration endpoint of the serum against cefotetan-coated RBCs was 40,960, while the serum plus soluble cefotetan against uncoated RBCs was 2,560. This case of severe cefotetan-induced hemolysis was complicated by an acute hemolytic event that occurred during the transfusion of PRBC. Clinical and transfusion service staff must consider drug-induced hemolysis in the differential diagnosis of acute anemia. PMID- 10815792 TI - Folate sufficient subjects do not accumulate additional folates during supplementation. AB - In a double-blinded placebo-controlled trial of folic acid supplementation in 82 alcoholic subjects, it was found that whole blood folate levels, determined by a mass spectrometric method, do not increase in subjects whose baseline folate levels are above the third quartile (folate sufficiency). Since a state of folate sufficiency can now be identified, a recommended daily allowance (RDA) for folate can be determined using objective means. PMID- 10815793 TI - Autoimmune thyroid disease and antiphospholipid antibodies. AB - OBJECTIVE: Autoimmune thyroid disease (ATD) is associated with circulating autoantibodies reactive with epitopes on thyroid tissue and that are thought to be pathogenic in the development of these diseases. Antiphospholipid antibodies (APLA) are a family of immunoglobulins that recognize a variety of plasma proteins in association with anionic phospholipids. These antibodies may lead to a number of clinical syndromes including venous and arterial thromboses, thrombocytopaenia, and recurrent fetal loss. We have studied the prevalence of APLA in patients with ATD and have determined the prevalence of the APLA syndrome among APLA-positive patients. DESIGN: The study was a retrospective survey of patients with autoimmune thyroid disease attending the endocrinology clinic of a tertiary care academic hospital. PATIENTS AND MEASUREMENTS: One hundred and thirty patients with autoimmune thyroid disease from the endocrinology clinic at our hospital were studied. 84% had chronic thyroiditis and 16% had Graves' disease. Free T4 and thyroid stimulating hormone (TSH) levels, antimicrosomal and antithyroglobulin antibodies, and an antiphospholipid antibody test were performed on all subjects. RESULTS: 43% of patients with chronic thyroiditis and 43% of patients with Graves' disease were APLA positive, with an overall rate of 43% APLA positivity among patients with ATD. Of the 56 patients that were APLA positive, forty-eight (86%) had APLA of the IgG subtype, four (7%) had IgM antibodies, and nine (16%) had both IgG and IgM antibodies. None of the patients had clinical evidence of the APLA syndrome. CONCLUSIONS: We conclude that the prevalence of APLA in ATD is increased compared to healthy individuals but that this is likely to be an epiphenomenon. However, prolonged follow up is necessary in order to determine the true clinical significance of these antibodies in ATD patients. PMID- 10815794 TI - Multiple myeloma presenting high fever and high serum levels of lactic dehydrogenase, CRP, and interleukin-6. AB - Two myeloma patients presented high fever with no signs or data indicating infection at diagnosis or relapse. Both patients had plasmablastic myeloma, and serum levels of lactic dehydrogenase (LDH) and CRP were extremely high. Plasmablastic morphology, high LDH, and CRP were recognized as poor prognostic factors, indicating a fulminant phase of multiple myeloma. Interleukin-6 (IL-6) was only high in measured cytokines. We proposed that IL-6 caused high fever and induced the fulminant phase in these 2 cases. PMID- 10815796 TI - Splenectomy in thrombotic thrombocytopenic purpura. PMID- 10815795 TI - Very severe aplastic anemia following resection of lymphocytic thymoma: effectiveness of antilymphocyte globulin, cyclosporin A, and granulocyte-colony stimulating factor. PMID- 10815797 TI - Bilateral central retinal artery occlusion secondary to sickle cell disease. PMID- 10815798 TI - Polycythemia in patients infected with human immunodeficiency virus-1. PMID- 10815799 TI - Activation of Stat3 preassembled with platelet-derived growth factor beta receptors requires Src kinase activity. AB - Members of the STAT family of transcriptional regulators modulate the expression of a variety of gene products that promote cell proliferation, survival and transformation. Although initially identified as mediators of cytokine signaling, the STAT proteins are also activated by, and thus may contribute to the actions of, polypeptide growth factors. To define the mechanism by which these factors activate STATs, we examined the process of Stat3 activation in Balb/c-3T3 fibroblasts treated with platelet-derived growth factor (PDGF). As STATs are activated by tyrosine phosphorylation, and as PDGF receptors are ligand-activated tyrosine kinases, we considered the possibility that Stat3 interacts with and is phosphorylated by PDGF receptors. We find that Stat3 associates with PDGF beta receptors in both the presence and, surprisingly, the absence of PDGF. Moreover, Stat3 was phosphorylated on tyrosine in PDGF beta receptor immunoprecipitates of PDGF-treated but not untreated cells. Although required, receptor activation was insufficient for Stat3 activation. When added to cells in combination with a pharmacologic agent (PD180970) that specifically inhibits the activity of Src family tyrosine kinases, PDGF did not activate Stat3 as monitored by electrophoretic mobility shift assay. PD180970 did not affect MAPK activation by PDGF or the JAK-dependent activation of Stat3 by interleukin-6. The necessity of Src activity for Stat3 activation by PDGF was further evidenced by data showing the presence of Src in complexes containing both Stat3 and PDGF beta receptors in PDGF-treated cells. These results suggest a novel mechanism of STAT activation in which inactive Stat3 pre-assembles with inactive PDGF receptors, and in response to ligand binding and in a manner dependent on Src kinase activity, is rapidly phosphorylated and activated. Additional data demonstrate that Src kinase activity is also required for PDGF stimulation of DNA synthesis in density arrested cells. PMID- 10815800 TI - IL-2 and long-term T cell activation induce physical and functional interaction between STAT5 and ETS transcription factors in human T cells. AB - Activation of Stat5 by many cytokines implies that it cannot alone insure the specificity of the regulation of its target genes. We have evidenced a physical and functional interaction between members of two unrelated transcription factor families, Ets-1, Ets-2 and Stat5, which could contribute to the proliferative response to interleukin 2. Competition with GAS- and EBS-specific oligonucleotides and immunoassays with a set of anti-Stat and anti-Ets families revealed that the IL-2-induced Stat5-Ets complex recognizes several GAS motifs identified as target sites for activated Stat5 dimers. Coimmunoprecipitation experiments evidenced that a Stat5/Ets-1/2 complex is formed in vivo in absence of DNA. GST-pull down experiments demonstrated that the C-terminal domain of Ets 1 is sufficient for this interaction in vitro. Cotransfection experiments in Kit225 T cells resulted in cooperative transcriptional activity between both transcription factors in response to a combination of IL-2, PMA and ionomycin. A Stat5-Ets protein complex was the major inducible DNA-binding complex bound to the human IL-2rE GASd/EBSd motif in long-term proliferating normal human T cells activated by CD2 and CD28. These results suggest that the inducible Stat5-Ets protein interaction plays a role in the regulation of gene expression in response to IL-2 in human T lymphocytes. PMID- 10815802 TI - All-trans-retinoic acid upregulates TNF receptors and potentiates TNF-induced activation of nuclear factors-kappaB, activated protein-1 and apoptosis in human lung cancer cells. AB - Retinoids modulate the growth and differentiation effects of TNF but the mechanism is not understood. In this study, we investigated the effect of all trans-retinoic acid (ATRA) on the cell surface expression of TNF receptors and receptor-mediated signaling in various human lung cancer cell lines. ATRA treatment of cells that express wild-type p53 (A549 and H460), or null p53 (H1299), or mutant p53 (H596) increased the number of TNF receptors, as determined by the specific binding of 125I-labeled TNF to these cells, in a dose- and time-dependent manner. Treatment with 2 microm ATRA for 24 h at 37 degrees C produced the maximal increase. Scatchard analysis indicated that the increase induced by ATRA was due to an increase in receptor number and not to an increase in affinity. The upmodulation of TNF receptors was also confirmed by covalent receptor-ligand cross-linking studies. The increase in TNF receptors sensitized H596 cells to TNF-induced activation of NF-kappaB, AP-1 and apoptosis. A549 cells, however, were completely resistant to TNF-induced activation of NF-kappaB, AP-1 and apoptosis. Treatment of these cells with as little as 0.5 microM ATRA was effective in converting TNF-resistant cells to TNF-sensitive. Overall our results indicate that ATRA induces the TNF receptors in human lung cancer cells, which sensitizes them to TNF-induced signaling leading to activation of NF kappaB, AP-1 and apoptosis. PMID- 10815801 TI - Evi27 encodes a novel membrane protein with homology to the IL17 receptor. AB - Evi27 is a common site of retroviral integration in BXH2 murine myeloid leukemias. Here we show that integration at Evi27 occurs in a CpG island approximately 6 kb upstream from a novel gene (designated Evii27) with homology to the IL17 receptor (Il17r) and that proviral integrations result in increased expression of the Evi27 protein on the cell surface. The human EVI27 homolog was also cloned and mapped to chromosome 3p21. Multiple Evi27 isoforms were detected at the RNA and protein level in both human and mouse, indicating that Evi27 expression is complex. Some of the isoforms are shown to likely represent secreted soluble forms of the protein produced by intron incorporation or by proteolytic cleavage. In the mouse, highest Evi27 expression occurs in liver and testes with lower expression in kidney and lung. In humans, EVI27 is expressed at high levels in the kidney, with moderate levels in the liver, brain, and testes. Within hematopoietic cells, Evi27 expression is restricted. Northern and Western analysis showed that Evi27 is expressed in selected T-cell, B-cell and myeloid cell lines. These results suggest that Evi27 expression is tightly regulated during hematopoietic differentiation. Collectively, these studies identify a new member of the cytokine receptor family whose increased and uncoordinated expression may lead to myeloid leukemia by altering Evi27's normal ability to control the growth and/or differentiation of hematopoietic cells. PMID- 10815803 TI - The secreted glycoprotein CREG enhances differentiation of NTERA-2 human embryonal carcinoma cells. AB - Differentiation of the human embryonal carcinoma cell line NTERA-2 is characterized by changes in morphology, altered patterns of gene expression, reduced proliferative potential, and a loss of tumorigenicity. The cellular repressor of E1A-stimulated genes, CREG, was previously shown to antagonize transcriptional activation and cellular transformation by the Adenovirus E1A oncoprotein. These properties suggested that CREG may function to inhibit cell growth and/or promote differentiation. Here we show that CREG is a secreted glycoprotein which enhances differentiation of NTERA-2 cells. Northern blot analysis reveals that, although CREG mRNA is widely expressed in adult tissues, CREG mRNA is not significantly expressed in pluripotent mouse embryonic stem cells or NTERA-2 embryonal carcinoma cells. CREG mRNA is rapidly induced upon in vitro differentiation of both mouse embryonic stem cells and human NTERA-2 cells. We show that constitutive expression of CREG in NTERA-2 cells enhances neuronal differentiation upon treatment with retinoic acid. Media enriched in CREG was also found to promote NTERA-2 differentiation in the absence of an inducer such as retinoic acid. These studies suggest that secreted CREG protein participates in a signaling cascade important for differentiation of pluripotent stem cells such as those found in teratocarcinomas. PMID- 10815804 TI - Mammary gland specific hEGF receptor transgene expression induces neoplasia and inhibits differentiation. AB - The epidermal growth factor receptor (EGFR) is overexpressed in about 48% of human breast cancer tissues. To analyse the role of the EGFR in mammary tumor development we generated transgenic mice expressing the human EGFR under the control of either the MMTV-LTR (MHERc) or the beta-lactoglobulin promoter (BLGHERc). The BLGHERc-transgene was expressed exclusively in the female mammary gland, whereas the MHERc transgene was expressed more promiscuously in other organs, such as ovary, salivary gland and testis. Female virgin and lactating transgenic mice of both strains have impaired mammary gland development. Virgin EGFR transgenic mice developed mammary epithelial hyperplasias, whereas in lactating animals progression to dysplasias and tubular adenocarcinomas was observed. In both strains the number of dysplasias increased after multiple pregnancies. The transgene expression pattern was heterogeneous, but generally restricted to regions of impaired mammary gland development. Highest EGFR transgene expression was observed in adenocarcinomas. By using a whole mount organ culture system to study the differentiation potential of the mammary epithelium, we observed a reduced number of fully developed alveoli and a decrease in whey acidic protein expression. Taken together, EGFR overexpression results in a dramatic effect of impaired mammary gland development in vitro as well as in vivo, reducing the differentiation potential of the mammary epithelium and inducing epithelial cell transformation. PMID- 10815806 TI - A dominant negative RAS-specific guanine nucleotide exchange factor reverses neoplastic phenotype in K-ras transformed mouse fibroblasts. AB - Ras proteins are small GTPases playing a pivotal role in cell proliferation and differentiation. Their activation state depends on the competing action of GTPase Activating Proteins (GAP) and Guanine nucleotide Exchange Factors (GEF). A tryptophan residue (Trp1056 in CDC25Mm-GEF), conserved in all ras-specific GEFs identified so far has been previously shown to be essential for GEF activity. Its substitution with glutamic acid results in a catalytically inactive mutant, which is able to efficiently displace wild-type GEF from p21ras and to originate a stable ras/GEF binary complex due to the reduced affinity of the nucleotide-free ras/GEF complex for the incoming nucleotide. We show here that this 'ras sequestering property' can be utilized to attenuate ras signal transduction pathways in mouse fibroblasts transformed by oncogenic ras. In fact overexpression of the dominant negative GEFW1056E in stable transfected cells strongly reduces intracellular ras-GTP levels in k-ras transformed fibroblasts. Accordingly, the transfected fibroblasts revert to wild-type phenotype on the basis of morphology, cell cycle and anchorage independent growth. The reversion of the transformed phenotype is accompanied by DNA endoreduplication. The possible use of dominant negative ras-specific GEFs as a tool to down-regulate tumor growth is discussed. PMID- 10815805 TI - Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells. AB - Vascular endothelial growth factor (VEGF) is a principal regulator of vasculogenesis and angiogenesis. VEGF expresses its effects by binding to two VEGF receptors, Flt-1 and KDR. However, properties of Flt-1 and KDR in the signal transduction of VEGF-mediated effects in endothelial cells (ECs) were not entirely clarified. We investigated this issue by using two newly developed blocking monoclonal antibodies (mAbs) against Flt-1 and KDR. VEGF elicits DNA synthesis and cell migration of human umbilical vein endothelial cells (HUVECs). The pattern of inhibition of these effects by two mAbs indicates that DNA synthesis is preferentially mediated by KDR. In contrast, the regulation of cell migration by VEGF appears to be more complicated. Flt-1 regulates cell migration through modulating actin reorganization, which is essential for cell motility. A distinct signal is generated by KDR, which influences cell migration by regulating cell adhesion via the assembly of vinculin in focal adhesion plaque and tyrosine-phosphorylation of focal adhesion kinase (FAK) and paxillin. PMID- 10815807 TI - HTLV-I Tax transrepresses the human c-Myb promoter independently of its interaction with CBP or p300. AB - The c-Myb proto-oncogene is preferentially expressed in hematopoietic lineages, and highly expressed in several leukemia types. The Human T-cell Leukemia Virus Type I (HTLV-I) is the etiological agent of Adult T-cell Leukemia/Lymphoma (ATLL). A previous report suggested that Tax, the viral transactivator, is able to suppress the transactivation potential of c-Myb protein by competing for recruitment of CBP. We tested whether such a competition could affect transcription from the c-Myb promoter in Tax expressing T-cells. Using several c Myb promoter reporter constructs carrying mutations in various regions, we demonstrate that Tax suppression of c-Myb transactivation results in transrepression of the c-Myb promoter through the Myb responsive elements in Jurkat T-cells. The ability of Tax mutants M22, M47 and V89A to interact with the full-length CBP and p300 proteins in vitro, and their ability to repress the c Myb promoter, was then evaluated. Although both M47 and M22 bind to CBP and p300 to a similar extent, only M47 was able to repress the c-Myb promoter, suggesting that competition for CBP/p300 binding was not the mechanism underlying Tax's effect. This concept was further supported by the fact that the Tax mutant V89A transrepresses the c-Myb promoter efficiently in spite of an impaired binding to CBP and p300. Therefore, Tax-mediated repression of the c-Myb promoter appears to be independent from a direct competition between c-Myb and Tax for recruitment of CBP/p300. Interestingly, a decreased transcription from the endogenous c-Myb promoter was observed in several HTLV-I transformed T-cell lines. Finally, the ability of Tax to directly repress the endogenous c-Myb promoter was demonstrated in a Jurkat cell line stably transfected with a tax gene driven by a cadmium inducible promoter. PMID- 10815808 TI - p21Waf1/Cip1/Sdi1-induced growth arrest is associated with depletion of mitosis control proteins and leads to abnormal mitosis and endoreduplication in recovering cells. AB - Induction of a cyclin-dependent kinase inhibitor p21Waf1/ Cip1/Sdi1 is an integral part of cell growth arrest associated with senescence and damage response. p21 overexpression from an inducible promoter resulted in senescence like growth arrest in a human fibrosarcoma cell line. After release from p21 induced growth arrest, cells re-entered the cell cycle but displayed growth retardation, cell death and decreased clonogenicity. The failure to form colonies was associated with abnormal mitosis and endoreduplication in the recovering cells and was correlated with the induced level of p21 and the duration of p21 induction. p21 induction was found to inhibit the expression of multiple proteins involved in the execution and control of mitosis. p21-induced depletion of the cellular pools of mitosis-control proteins was followed by asynchronous resynthesis of such proteins after release from p21, which explains the observed mitotic abnormalities. Genetic destabilization in cells recovering from p21 induced growth arrest may conceivably play a role in carcinogenesis and tumor progression. PMID- 10815809 TI - Anatomy and the art of Archaic Greece. PMID- 10815810 TI - Animating functional anatomy for the web. AB - The instructor sometimes has a complex task in explaining the concepts of functional anatomy and embryology to health professional students. However, animations can easily illustrate functional anatomy, clinical procedures, or the developing embryo. Web animation increases the accessibility of this information and makes it much more useful for independent student learning. A modified version of the animation can also be used for patient education. This article defines animation, provides a brief history of animation, discusses the principles of animation, illustrates and evaluates some of the video-editing or movie-making computer software programs, and shows examples of two of the author's animations. These two animations are the inferior alveolar nerve block from the mandibular nerve anesthetics unit and normal temporomandibular joint (TMJ) function from the muscles of the mastication and the TMJ function unit. The software discussed are the industry leaders and have made the job of producing computer-based animations much easier. The programs are Adobe Premiere, Adobe After Effects, Apple QuickTime and Macromedia Flash . PMID- 10815811 TI - Going virtual with quicktime VR: new methods and standardized tools for interactive dynamic visualization of anatomical structures. AB - Continuing evolution of computer-based multimedia technologies has produced QuickTime, a multiplatform digital media standard that is supported by stand alone commercial programs and World Wide Web browsers. While its core functions might be most commonly employed for production and delivery of conventional video programs (e.g., lecture videos), additional QuickTime VR "virtual reality" features can be used to produce photorealistic, interactive "non-linear movies" of anatomical structures ranging in size from microscopic through gross anatomic. But what is really included in QuickTime VR and how can it be easily used to produce novel and innovative visualizations for education and research? This tutorial introduces the QuickTime multimedia environment, its QuickTime VR extensions, basic linear and non-linear digital video technologies, image acquisition, and other specialized QuickTime VR production methods. Four separate practical applications are presented for light and electron microscopy, dissectable preserved specimens, and explorable functional anatomy in magnetic resonance cinegrams. PMID- 10815812 TI - Web-delivery of anatomy video clips using a CD-ROM. AB - Until recently, anatomists had no doubt that the teaching of anatomy had to include cadaver dissection. However, due to a changing academic environment as well as challenged financial institutional resources, computer-assisted instruction was introduced into medical curriculum in an attempt to reduce the cost and the time committed to cadaver dissection. Computer-assisted instruction included locally created or commercially available anatomy software, Internet sites, and databases of digital images of cadaveric structures such as the Virtual Human Project. However, until now, bandwidth limitations have not allowed effective visualization in real-time over the Internet of recorded videos or 3D animations reconstructed from a database. We describe how to successfully link and display large video clips stored on a CD-ROM in support of lectures saved in HTML format on the Internet. This process, described in its totality, allows students to access audiovisual files on a CD-ROM through the Internet, from any location, with either Macintosh or Windows computers, using the Netscape browser. This process allowed us to circumvent one of the most significant limitations of the computer-assisted instruction on the Internet by delivering full audio and visual information on demand, as it would happen in a traditional classroom. PMID- 10815813 TI - Animated PowerPoint as a tool to teach anatomy. AB - Anatomy is a visual science. For centuries, anatomic information has been conveyed through drawings that have been presented to students through every available medium. The projection of animated images from a computer is a medium that offers great promise in effecting improved communication of anatomic information. Using Microsoft PowerPoint software, we have developed animated presentations for all of our lectures in Gross and Developmental Anatomy. As a starting point, we scan pen-and-ink drawings to create a digital image. The image may be edited and manipulated in an image processing program. Next, the image is imported into a PowerPoint slide where it is labeled and otherwise enhanced (arteries overlaid with red color, veins in blue, etc.) and the enhancements are animated, as we describe here step by step. For the lecture, the file is loaded on a server that is accessible through a network from a computer in the lecture hall. The output is directed to a video projector and the PowerPoint presentation is projected in the "Slide Show" mode. We use a wireless mouse that allows us to control the presentation from anywhere in the room. Before the lecture, students are provided with the same unlabeled drawings as handouts, and during the lecture the students are actively engaged in labeling the drawings and making related notes. After the lecture, the file is saved in HTML format and posted on our course web site where students can access the slides. Evaluation by the students at the end of the course demonstrated that this style of presentation was very favorably received. PMID- 10815814 TI - Gross anatomy of the head and neck and neuroscience in an integrated first-year medical school curriculum. AB - The curriculum for first year medical students at the University of Cincinnati has changed. Beginning in the fall of 1998, material in the first year was presented in an Integrated Educational Program. The goal of this program was to provide students with an understanding of the normal structure, function, and development of the human body. The purpose of this report is to discuss the unique integration that occurs in a block offered during the Spring Quarter. The two components of this block are Gross Anatomy of the Head and Neck and Brain and Behavior I. Brain and Behavior I is a new offering combining neuroanatomy, neurophysiology, neurology, and a psychiatry/behavioral component. The unique combinations offered in this block are logical, educationally sound, and have been enthusiastically received by both the students and faculty. PMID- 10815815 TI - Relationship between quadriceps strength and rate of loading during gait in women. AB - One function of skeletal muscle is to serve as the body's shock absorbers and thus dampen rates of loading during activity. The aim of this cross-sectional study was to determine the significance of muscle strength on rates of loading during gait. Thirty-seven women (mean age: 34.5 +/- 8.2 years) were solicited by advertisement and placed into one of two groups-strength-trained or sedentary-on the basis of training history. They walked (10 trials) over a 10-m walkway at a controlled speed of 1.22-1.35 m/s while the rate of loading was sampled with a 1,000-Hz force platform. Quadriceps and hamstring strength was measured at 90 degrees/s with an isokinetic dynamometer. Statistical analyses (p < 0.05) included descriptive statistics and unpaired t tests for comparison between groups. The women in the sedentary group weighed more and had significantly less concentric and eccentric strength of the quadriceps and hamstrings relative to body weight than did those in the strength-trained group. In addition, they demonstrated significantly higher rates of loading (2.21 +/- 0.15 compared with 1.75 +/- 0.08%wt/ms) than those in the strength-trained group. PMID- 10815816 TI - Significance of changes in the reference position for measurements of tibial translation and diagnosis of cruciate ligament deficiency. AB - Measurements of tibial translation in response to an external load are used in clinical and laboratory settings to diagnose and characterize knee-ligament injuries. Before these measurements can be quantified, a reference position of the knee must be established (defined as the position of the knee with no external forces or moments applied). The objective of this study was to determine the effects of cruciate ligament deficiency on this reference position and on subsequent measurements of tibial translation and, in so doing, to establish a standard of kinematic measurement for future biomechanical studies. Thirty-six human cadaveric knees were studied with a robotic/universal force-moment sensor testing system. The reference positions of the intact and posterior cruciate ligament-deficient knees of 18 specimens were determined at full extension and at 30, 60, 90, and 120 degrees of flexion, and the remaining five-degree-of-freedom knee motion was unrestricted. Subsequently, under a 134-N anterior-posterior load, the resulting knee kinematics were measured with respect to the reference positions of the intact and posterior cruciate ligament-deficient knees. With posterior cruciate ligament deficiency, the reference position of the knee moved significantly in the posterior direction, reaching a maximal shift of 9.3 +/- 3.8 mm at 90 degrees of flexion. For the posterior cruciate ligament-deficient knee, posterior tibial translation ranged from 13.0 +/- 3.4 to 17.7 +/- 3.6 mm at 30 and 90 degrees, respectively, when measured with respect to the reference positions of the intact knee. When measured with respect to the reference positions of the posterior cruciate ligament-deficient knee, these values were significantly lower, ranging from 11.7 +/- 4.3 mm at 30 degrees of knee flexion to 8.4 +/- 4.8 mm at 90 degrees. A similar protocol was performed to study the effects of anterior cruciate ligament deficiency on 18 additional knees. With anterior cruciate ligament deficiency, only a very small anterior shift in the reference position was observed. Overall, this shift did not significantly affect measurements of tibial translation in the anterior cruciate ligament-deficient knee. Thus, when the tibial translation in the posterior cruciate ligament injured knee is measured when the reference position of the intact knee is not available, errors can occur and the measurement may not completely reflect the significance of posterior cruciate ligament deficiency. However, there should be less corresponding error when measuring the tibial translation of the anterior cruciate ligament-injured knee because the shift in reference position with anterior cruciate ligament deficiency is too small to be significant. We therefore recommend that in the clinical setting, where the reference position of the knee changes with injury, comparison of total anterior-posterior translation with that of the uninjured knee can be a more reproducible and accurate measurement for assessing cruciate-ligament injury, especially in posterior cruciate ligament-injured knees. Similarly, in biomechanical testing where tibial translations are often reported for the ligament-deficient and reconstructed knees, a fixed reference position should be chosen when measuring knee kinematics. If such a standard is set, measurements of knee kinematics will more accurately reflect the altered condition of the knee and allow valid comparisons between studies. PMID- 10815817 TI - Characterization of hamstring reflexes during anterior cruciate ligament disruption: in vivo results from a goat model. AB - The existence of an anterior cruciate ligament-hamstring reflex arc, the extent to which these reflexes can protect the knee, and the extent to which they are affected by rupture of the anterior cruciate ligament remain controversial. We evaluated the temporal components of the anterior cruciate ligament-hamstring synergy by simulating an injury to the ligament in a goat model. Reflexive hamstring activation in anesthetized goats was evaluated when the anterior cruciate ligament was loaded with static subfailure, dynamic subfailure, and dynamic failure loads. Reflexive hamstring activation was not found in response to static subfailure loading but was observed in response to dynamic subfailure and failure loading. The latency of the reflex evoked by dynamic failure loading was shorter than that evoked by dynamic subfailure loading. The findings suggest that the extent to which the hamstring reflexes can protect the knee may be bounded by the ability of these muscles to generate force rapidly and the amplitude and time-course of the loads applied to the knee joint. The present data present a framework for further investigation of the contribution of anterior cruciate ligament-hamstring reflexes to the stability of the knee joint under high loads and loading rates. PMID- 10815818 TI - In vivo technique to quantify the internal-external rotation kinematics of the human glenohumeral joint. AB - Internal and external rotation of the humerus are often related to instability, injury mechanisms, and surgical and rehabilitation outcomes at the glenohumeral joint. The goal of this study was to develop a technique to quantify the internal external rotation kinematics of the glenohumeral joint in human subjects, including the rotational range of motion, neutral-zone laxity, and flexibility. For both arms of 10 normal subjects, the rotational range of motion of the humerus was assessed at 45 degrees of abduction with 4 Nm of applied moment to produce internal and external rotations about the long axis. The neutral zone was defined as the portion of the rotational range of motion that occurred between +1 and -1 Nm of applied internal-external rotation torque. The flexibility was determined from the slope of the moment-rotation curve from 1 to 4 Nm of applied moment. The repeatability of the device during two trials on the same day and two trials 1 week apart was determined. There were no significant differences between the two same-day and two across-day trials for each outcome measure. The internal external rotational range of motion was 139.4 degrees (SD 40.5 degrees). The neutral-zone laxity was 77.8 degrees (SD 46.0 degrees). With a linear approximation, the external rotation flexibility (20.1 degrees/Nm [SD 13.7 degrees/Nm]) was four times greater than the internal rotation flexibility (5.8 degrees/Nm [SD 5.1 degrees/Nm]). The changes in the magnitude of the laxity, the ratio between the laxity and the range of motion, or the values for flexibility determined with this technique could be used to describe joint laxity, surgical outcome, and rehabilitation progress. PMID- 10815819 TI - Assessment of elbow joint kinematics in passive motion by electromagnetic motion tracking. AB - This research provides a detailed analysis of the kinematics of passive elbow motion. It quantifies how closely humeroulnar kinematics approximates rotation around a fixed axis. The results are clinically relevant for emerging treatment modalities that impose an artificial hinge to the elbow joint, such as total elbow arthroplasty and articulated external fixation. In a cadaveric study of seven specimens, we quantified ulnar rotation around the humerus in terms of instantaneous screw displacement axes calculated from electromagnetic motion tracking source data. This methodology enabled description of the complex excursion of the elbow axis in terms of translation and orientation changes of the screw displacement axes over the range of motion. Furthermore, we analyzed the envelope of joint laxity for elbow motion under applied small varus and valgus moments. In addition, radiographic landmarks of clinical utility for axis location were evaluated by visualizing the elbow's radiographic appearance when viewed from along the calculated best-fit (average) rotation axis. Over the normal range of elbow motion, the screw displacement axis varied 2.6-5.7 degrees in orientation and 1.4-2.0 mm in translation. All instantaneous rotation axes nearly intersected on the medial facet of the trochlea. The breadth of the envelope of varus-valgus joint laxity was greatest within the initial 40 degrees of flexion and decreased by a factor of approximately two for flexion angles exceeding 100 degrees. PMID- 10815820 TI - Titanium particles inhibit osteoblast adhesion to fibronectin-coated substrates. AB - To illuminate the effect of titanium particles on osteoblast function, we compared the adhesion force of neonatal rat calvarial osteoblasts on fibronectin coated glass after incubation with titanium particles (80% had diameters of less than 5 microm). The cells were incubated with the particles for 1.5-72 hours. Using a micropipette single-cell manipulation system, we showed that the adhesion force of the osteoblasts to fibronectin-coated glass (1.0 microg/ml) was significantly affected by the presence of particulate debris. The adhesion force of the cells incubated with titanium particles for less than 4 hours was not significantly affected by exposure to the particles; after 4 hours, however, it was significantly reduced relative to that of controls. Aspiration of particle challenged osteoblasts into the micropipette demonstrated that the particles were not stripped from the cell surface and therefore confirmed that the osteoblasts had ingested them. During aspiration, the particles traveled through the cytoplasm rather than on the cell surface. When the osteoblasts were exposed to the particles and cytochalasin D, they exhibited much lower adhesion forces than did the controls or the cells exposed to titanium particles only; this indicates an important role of actin filaments in the osteoblastic response to particles. Staining for F-actin also indicated an influence of internalized titanium particulate on cytoskeletal arrangement and cell spreading. Furthermore, with standard Northern blotting techniques, levels of mRNA for collagen type I and fibronectin were significantly reduced as early as 4 hours after exposure to particles compared with levels in controls, and this effect continued to 72 hours. These data indicate that direct exposure of osteoblasts to titanium particles, which we propose to be ingested by the osteoblasts, can significantly decrease osteoblast adhesion force; this may lead to decreased cellular activity and gene expression of fibronectin and collagen type I in the presence of titanium wear debris. PMID- 10815821 TI - Adhesion formation can be reduced by the suppression of transforming growth factor-beta1 activity. AB - Surgery or trauma often results in restrictive adhesions around joints or tendons that cause severe functional impairment. The formation of adhesion is essentially a fibrogenetic process; therefore, peptide growth factors, such as transforming growth factor-beta, are assumed to play central roles in its development. The purpose of this study was to test the hypothesis that suppression of transforming growth factor-beta1 activity reduces adhesion formation. Sixty rabbits were prepared and randomly divided into six groups of 10. Intraarticular adhesions were created in the right knee joints by cortical bone shaving and subsequent cast immobilization for 4 weeks. In animals in three of the six groups, transforming growth factor-beta1 activity was suppressed by continuous administration of the neutralizing antibody in three graded doses; animals in the other three groups were used as controls. Four weeks after the surgery, the casts were removed and the adhesions were assessed macroscopically, histologically, biomechanically, and biochemically. Gross observation showed that the neutralizing antibody had suppressed adhesion formation in a dose-dependent manner. This is consistent with biomechanical measurement results demonstrating that the antibody reduced the flexion contractures. Histologically, the adhesion in our model was fibrous tissue and the adhesions in the animals in the antibody groups were thin and loose in comparison with the controls. Biochemical analyses further supported these results, demonstrating that administration of the antibody reduced collagen content in the adhesions with a predominance of type-I collagen. Thus, this study showed that suppression of the actions of transforming growth factor-beta1 reduced adhesion formation. Considering the various possible measures to control the activity of the growth factor, suppression of transforming growth factor-beta may be a novel, potent approach to preventing adhesions. PMID- 10815822 TI - Time-dependent increases in type-III collagen gene expression in medical collateral ligament fibroblasts under cyclic strains. AB - Numerous studies have demonstrated the capacity of mechanical strains to modulate cell behavior through several different signaling pathways. Understanding the response of ligament fibroblasts to mechanically induced strains may provide useful knowledge for treating ligament injury and improving rehabilitation regimens. Biomechanical studies that quantify strains in the anterior cruciate and medial collateral ligaments have shown that these ligaments are subjected to 4-5% strains during normal activities and can be strained to 7.7% during external application of loads to the knee joint. The objective of this study was to characterize the expression of types I and III collagen in fibroblast monolayers of anterior cruciate and medial collateral ligaments subjected to equibiaxial strains on flexible growth surfaces (0.05 and 0.075 strains) by quantifying levels of mRNA encoding these two proteins. Both cyclic strain magnitudes were studied under a frequency of 1 Hz. The results indicated marked differences in responses to strain regimens not only between types I and III collagen mRNA expression within each cell type but also in patterns of expression between anterior cruciate and medial collateral ligament cells. Whereas anterior cruciate ligament fibroblasts responded to cyclic strains by expression of higher levels of type-I collagen message with almost no significant increases in type-III collagen, medial collateral ligament fibroblasts exhibited statistically significant increases in type-III collagen mRNA at all time points after initiation of strain with almost no significant increases in type-I collagen. Furthermore, differences in responses by fibroblasts from the two ligaments were detected between the two strain magnitudes. In particular, 0.075 strains induced a time-dependent increase in type-III collagen mRNA levels in medial collateral ligament fibroblasts whereas 0.05 strains did not. The strain-induced changes in gene expression of these two collagens may have implications for the healing processes in ligament tissue. The differences may explain, in part, the healing differential between the anterior cruciate and medial collateral ligaments in vivo. PMID- 10815823 TI - Use of roentgenography and magnetic resonance imaging to predict meniscal geometry determined with a three-dimensional coordinate digitizing system. AB - To evaluate and improve on the procedures used by a tissue bank in selecting donor menisci for transplantation, this study was designed to fulfill four objectives: (a) define and quantify a set of independent parameters for describing the geometry of the medial and lateral menisci, (b) determine how well the sizing protocol of the tissue bank (i.e., two transverse roentgenographic measurements obtained from the injured knee or six transverse magnetic resonance imaging measurements obtained from the contralateral knee) predicts the four standard transverse parameters of the menisci, (c) determine if including one additional transverse roentgenographic measurement for each compartment improves the ability of roentgenograms to predict transverse meniscal parameters, and (d) determine if five magnetic resonance imaging measurements at three different meniscal cross sections of the contralateral knee predict the 15 standard cross sectional parameters of the meniscus in the injured knee. A laser-based, noncontacting three-dimensional coordinate digitizing system was used to determine surface coordinates from which menisci were reconstructed in a computer. For each reconstructed meniscus, four parameters in the transverse plane and five cross-sectional parameters in each of three regions (i.e., anterior, middle, and posterior) were defined, yielding a set of 19 standard parameters to describe the geometry. Through a correlation analysis, these standard parameters were shown to be largely unrelated to one another, thus confirming that the parameters form an independent set describing the three dimensional geometry of the menisci. The two roentgenographic measurements were poor predictors of transverse standard meniscal parameters, predicting only one of four standard parameters for the medial meniscus and none of four standard parameters for the lateral meniscus with coefficients of determination greater than or equal to 0.5. Including one additional roentgenographic measurement to the tissue bank protocol increased the number of standard transverse parameters predicted to three of four for the medial meniscus and two of four for the lateral meniscus. Magnetic resonance imaging was better than roentgenography for predicting the three-dimensional meniscal geometry. The transverse measurements from magnetic resonance imaging predicted three of four standard transverse parameters for the medial meniscus and all four for the lateral meniscus. With the addition of the cross-sectional measurements by magnetic resonance imaging, seven of 15 standard cross-sectional parameters were predicted for both the medial and lateral menisci. Assuming that a successful clinical outcome depends on how well an allograft matches the size and shape of the original meniscus, magnetic resonance imaging rather than roentgenography should be used for allograft size-matching by tissue banks. PMID- 10815824 TI - Early medial collateral ligament scars have inferior creep behaviour. AB - Recent evidence suggests that ligaments are subject to repetitive loads in vivo. Hence, the creep behaviour (increase in strain under constant or repetitive stress) of ligament scars is of significance, since healing ligaments may elongate permanently over time. A rabbit medial collateral ligament model was used to assess the creep behaviour of healing ligaments at stresses corresponding to 30% of the scar failure strength at 3 (n = 6), 6 (n = 6), and 14 (n = 5) weeks of healing. The stresses for the creep tests of scars (and contralateral controls) were 2.2, 4.1, and 7.1 MPa for the 3, 6, and 14-week healing intervals, respectively. Normal medial collateral ligaments from comparable rabbits were tested at two of the corresponding stresses: 4.1 (n = 7) and 7.1 (n = 6) MPa. Total creep strain-the cumulative increase in strain resulting from serial cyclic and static creep testing-was independent of the order of testing and was compared between scars and controls. Water contents after testing were also quantified. Water contents before testing were assessed for additional animals: six normal animals and three from each healing interval. At 3 weeks of healing, the total creep strain of medial collateral ligament scars was four times greater than that for contralateral controls tested at the same stress. Although there was improvement from 3 to 14 weeks, the total creep strain of scars remained more than two times greater than that of controls at 14 weeks. Scar water content decreased with healing from elevated initial values, possibly contributing to the marginally improved creep response. Comparisons of this deficiency in scar creep with previously published scar abnormalities in the same model suggest that collagen crosslink density, proteoglycan content, soft-tissue flaws, and the combined effect of collagen fibre changes may be mechanistic factors involved in scar creep. PMID- 10815825 TI - Expression of mRNA for vascular endothelial growth factor at the repair site of healing canine flexor tendon. AB - Neovascularization is an important and prominent feature of tendon healing that contributes to wound repair and potentially to adhesion formation. To define the location of cell populations that recruit and organize the angiogenic response during early healing of flexor tendon, we examined the gene expression pattern of the prototypic angiogenic factor, vascular endothelial growth factor, at and around the tenorrhaphy site in a canine model of flexor tendon repair. In situ hybridization with radiolabeled antisense riboprobes was used to identify tendon cell populations that contribute to the neovascularization process by expressing vascular endothelial growth factor and to relate this cell population to the previously described cell populations that participate in matrix synthesis (express type alpha1(I) collagen) and mitotic renewal (express histone H4). The majority of cells (approximately 67%) within the repair site itself express vascular endothelial growth factor mRNA; however, minimal levels accumulate within cells of the epitenon (approximately 10% of cells; p < 0.0002). By contrast, expression of type alpha1(I) collagen and histone H4 does not differ significantly between the epitenon and the repair site (uniformly approximately 30% of cells). Thus, a gradient of cell populations expressing vascular endothelial growth factor exists in the repairing tendon. These data suggest a potential contribution of cells within the repair site to the organization of angiogenesis during the early postoperative phase of tendon healing. PMID- 10815826 TI - MET oncogene aberrant expression in canine osteosarcoma. AB - The objective of this study was to investigate the role of the MET oncogene in canine osteosarcoma. Seven large-breed dogs affected by spontaneous skeletal osteosarcoma underwent en bloc tumor excision. Total RNA was extracted from frozen tumor samples and assessed for expression of the MET oncogene by Northern blot analysis. Five of seven biopsy samples expressed high levels of the MET oncogene; its expression in the primary tumors was comparable with that previously identified in primary osteosarcomas in humans. A lung metastasis from one of the dogs expressed MET at a higher level than did its primary tumor. Spontaneously arising osteosarcoma in dogs clinically and pathologically mimics the corresponding disease in humans. We previously demonstrated that the MET oncogene was aberrantly expressed in a high percentage of human osteosarcomas. The results of the current study also provide a molecular parallel between the tumors in dogs and humans. This in vivo model may be helpful in evaluating new strategies for therapy against osteosarcoma. PMID- 10815827 TI - Mechanical compression of the lumbar nerve root alters pain-related behaviors induced by the nucleus pulposus in the rat. AB - The purpose of this study was to refine a method of nerve-root injury in the rat to produce hyperalgesia, a pain-related behavior, and to determine if there were any relationships between the histological extent of nerve-root injury and the magnitude of hyperalgesia. Three methods were used to produce hyperalgesia: irritation of a nerve root by ectopic nucleus pulposus, silk loop alone, or both silk loop and ectopic nucleus pulposus. Autologous nucleus pulposus obtained from coccygeal intervertebral discs was relocated on the lumbar nerve roots after laminectomy. Two loops of 4-0 silk were placed around the exposed nerve roots. Hyperalgesia was measured preoperatively and postoperatively. The distribution of myelinated axons in the dorsal nerve roots was evaluated histologically. Mechanical hyperalgesia was detected in rats in which autologous nucleus pulposus was applied to the nerve root but not in those in which silk loops were used. Silk loops around the nerve root resulted in thermal hyperalgesia only in rats in which autologous nucleus pulposus was applied to the nerve root. Fewer large myelinated fibers were seen in the rats in which silk loops were used. Although a silk loop around the nerve root was not sufficient to produce hyperalgesia, supplemental application of autologous nucleus pulposus to the nerve root produced thermal hyperalgesia. It is possible that mechanical constriction of the nerve root alters the pain-related behavior elicited by chemical factors from the nucleus pulposus. PMID- 10815828 TI - Ability of osteoclast formation from peripheral monocytes using anti-fusion regulatory protein-1/CD98/4F2 monoclonal antibodies in patients with osteoporosis. AB - We investigated the difference in osteoclast formation between patients with osteoporosis and two healthy control groups by inducing it from peripheral blood monocytes with use of anti-fusion regulatory protein- monoclonal antibody. The group of patients with osteoporosis consisted of 35 women and excluded secondary osteoporosis, and the control groups consisted of 12 young healthy volunteers (control I) or 10 individuals age-matched to the patients with osteoporosis (control II). Osteoclast formation declined with age between the two control groups, but this decline was not significant. Fusion rate and the mean number of nuclei in osteoclasts were significantly less in the patients with osteoporosis than in the young or age-matched controls. It was clearly demonstrated that the ability of monocytes to fuse declines significantly in patients with osteoporosis. PMID- 10815829 TI - Longitudinal characterization of synovial fluid biomarkers in the canine meniscectomy model of osteoarthritis. AB - Damage to the meniscus can lead to posttraumatic osteoarthritis. Early markers of joint injury and tissue disease may be useful in developing and administering clinical treatment. We investigated the effects of total medial meniscectomy on biomarkers measured serially in synovial lavage fluid each month for 3 months. Following meniscectomy in dogs, four biomarkers were evaluated: cartilage oligomeric matrix protein, keratan sulfate epitope (5D4), the 3B3(-) neoepitope of chondroitin-6-sulfate, and the 3B3(+) chondroitinase-generated epitope of chondroitin-6-sulfate. Meniscectomy led to statistically significant elevations of all four biomarkers, with levels peaking at 4 weeks. By 12 weeks, the level of the 5D4 epitope returned to the preoperative baseline level whereas that of cartilage oligomeric matrix protein, 3B3(-), and 3B3(+) remained above the baseline. Concentrations of these biomarkers in the knees not operated on did not change significantly from the baseline. The levels of cartilage oligomeric matrix protein and 3B3(-) relative to 3B3(+) remained constant in all knees. In contrast, the level of 5D4 relative to 3B3(+) declined over time in the knee operated on but remained constant in the knee not operated on. These results demonstrate a quantitative change in the molecular components of synovial fluid after meniscectomy, as well as a qualitative change evinced by an alteration in the relative proportions of these epitopes. Extensive analyses showed a strong correlation between serum levels of 3B3(-) from the femoral and cephalic veins; however, serum 3B3(-) was not correlated with synovial fluid 3B3(-). These findings support the hypothesis that the concentrations of select cartilage biomarkers in synovial fluid are altered following meniscectomy and are promising tools for objectively monitoring the induction of osteoarthritis in this model system. PMID- 10815830 TI - Increased concentrations of nitrate and nitrite in the cyst fluid suggesting increased nitric oxide synthesis in solitary bone cysts. AB - The etiology and treatment of a solitary bone cyst have remained undefined. Surgical treatments have not been encouraging, because a less invasive corticosteroid-injection treatment has afforded good results. However, there has been little scientific rationale supporting corticosteroid treatment. In recent reports, bone-resorbing factors, including matrix metalloproteinases, prostaglandins, interleukin-1, and oxygen free radicals, have been demonstrated in the cyst fluid. To better elucidate the pathophysiology of the solitary bone cyst, we examined the activities of nitric oxide and cytokines in the cyst fluid as well as in the cyst membrane. The levels of nitrate and nitrite were significantly higher in the cyst fluid than in serum. Immunostaining of cells in the stroma and lining cells of the cyst wall was strongly positive for inducible nitric synthase. The levels of interleukin-6 and interleukin-1beta in the cyst fluid were elevated, and cells in the cyst membrane were positive for tumor necrosis factor-alpha, interleukin-6, and interleukin-1beta. Cultured cells from the cyst membrane were induced in the production of nitrate and nitrite in response to cytokine treatment. These findings suggest that the solitary bone cyst was in a state favorable for the production of nitric oxide. PMID- 10815831 TI - Evaluation of recombinant human bone morphogenetic protein-2 as a bone-graft substitute in a canine segmental defect model. AB - A study was performed in dogs to evaluate the dose-response characteristics and effectiveness of recombinant human bone morphogenetic protein-2 with a collagen sponge carrier in a segmental defect model. Twenty-seven dogs underwent bilateral radial osteotomies with creation of a 2.5-cm diaphyseal defect. All received autogenous cancellous bone graft in one defect and a collagen implant in the other. These implants contained recombinant human bone morphogenetic protein-2 at the following doses: group 1 at 0 microg (three dogs, 0 microg/ml total implant volume), group 2 at 150 microg (three dogs, 50 microg/ml), group 3 at 600 ,g (three dogs, 200 microg/ml), group 4 at 2,400 microg (three dogs, 800 microg/ml), group 5 at 0 microg (five dogs, 0 microg/ml), group 6 at 150 microg (five dogs, 200 microg/ml), and group 7 at 600 microg (five dogs, 50 microg/ml). The defects were stabilized with external fixators. The dogs in groups 1-4 were killed at 12 weeks postoperatively, and those in groups 5-7 were killed at 24 weeks postoperatively except for one dog in group 7, which was killed at 48 weeks. Evaluation included monthly radiographs, biomechanical testing, and nondemineralized histology. All 27 radii with autogenous cancellous bone graft and all 19 implants treated with recombinant human bone morphogenetic protein-2 achieved radiographic and histologic union and gross stability. The eight radii treated with collagen carrier alone went on to radiographic and histologic nonunion and were grossly unstable at death. A dose-dependent occurrence of cyst like bone voids was noted radiographically and histologically. Biomechanical performance tended to be better at the lowest dose studied at 12 weeks, and all three doses performed better than the placebo (p < 0.05) at 12 and 24 weeks. By 24 weeks, radiolucent areas corresponding to histologic bone voids persisted radiographically, although there was evidence of early bone remodeling. This remodeling progressed to 48 weeks in the single animal followed to this time point, although bone voids remained. These radiologic findings were confirmed histologically. Recombinant human bone morphogenetic protein-2 in a collagen sponge carrier has significant osteoinductive activity in this canine segmental defect model. A dose-response relationship is evident, with heterotopic bone and cyst-like void formation at higher doses and a minimum effective dose of 0-150 microg. At 12 and 24 weeks postoperatively, biomechanical parameters achieved by defects treated with recombinant human bone morphogenetic protein-2 were comparable with those of autograft controls and were significantly stronger than those of the placebo (p < 0.05). PMID- 10815832 TI - Experimental model for allograft incorporation and allograft fracture repair. AB - This study describes a rat model of allograft osteotomy healing. An intercalary skeletal defect was created in adult Lewis rats by resecting a 2-cm segment of the femur in the diaphysis, including the periosteum and the cuff of muscle layers. The skeletal defects were replaced with fresh-frozen devascularized intercalary allografts from Sprague-Dawley rats. A transverse osteotomy was made in the middle of the allograft. The osteotomized segments were stabilized with an intramedullary threaded Kirschner wire, which allowed immediate ambulation. Radiographic and histological examination at 4 and 8 weeks revealed a characteristic healing process at three different interfaces. Radiographically, the distal metaphyseal host-donor junction healed faster than the proximal diaphyseal host-donor interface. The osteotomy site did not have evidence of an intramembranous or endochondral repair process. This model can serve as a baseline for assessing allograft incorporation and fracture repair. PMID- 10815833 TI - Mechanical performance of standard and cannulated 4.0-mm cancellous bone screws. AB - The mechanical performance of bone screws is determined by their pull-out strength (holding power), compressive force, stripping torque, yield bending moment, ultimate bending moment, and fatigue strength. These parameters are related to the parameters of the screw design, including major thread diameter, minor thread diameter, thread length, pitch, shaft diameter, cannulation diameter, and material properties. The goal of the study was to theoretically predict the static performance of five 4.0-mm, 45-46-mm-long, cancellous, partially threaded standard and cannulated bone screws and compare the predictions with experimental measurements. A secondary goal was to determine if cannulation of the bone screw diminished its mechanical performance. The predicted values for pull-out force, compressive force, and stripping torque were determined by the thread length, major thread diameter, and thread shape factor. The screws with the largest major thread diameter and longest thread length had the greatest pull-out force, compressive strength, and stripping torque. However, when correcting for the thread length, a higher thread shape factor compensated for a smaller major diameter. The coefficient of determination (r2) for the correlation between the predicted and measured pull-out force improved from 0.75 to 0.90 when the theoretical model included the thread shape factor. The yield and ultimate bending moments are a function of the section modulus and material properties of the screw. The Ace solid screw had the greatest section modulus and yield and ultimate bending moments. The experimental data support the theoretical models for predicting the mechanical performance of bone screws. The design of the bone screws can be optimized on the basis of theoretical modeling. The strong correlation between the predicted and measured parameters allows comparison between bone screws without repeated experimental tests. Theoretical and experimental results show that cannulation of the bone screw did not inherently diminish its mechanical performance. PMID- 10815834 TI - Efficiency of systematic sampling in histomorphometric bone research illustrated by hydroxyapatite-coated implants: optimizing the stereological vertical-section design. AB - The sampling efficiency of the unbiased stereological vertical-section method was analyzed in five hydroxyapatite-coated implants. They were inserted into humans and harvested after 1 year. To find an optimal sampling design for histomorphometric analyses, sampling efficiency was estimated by variance analyses at different sampling levels (humans, sections, fields of view, and number of counting items) and intensities. Only minor changes in variance were observed when the initial scheme was reduced to include just one of the two possible implant sides, every third field of view, and half the density of the probe; this reduced the total workload at the microscope to less than 10% for all sections. In addition, the number of sections for analysis could be reduced to every fourth section per implant (three to four sections for evaluation) without significantly increasing variance. The study demonstrated that biological variation contributed to the majority of the total observed variance. Optimizing the sampling design could significantly reduce the workload at the hard-tissue microtome and the microscope without reducing the quality of the data that were unbiased and that had low sampling variance as compared with the true biological variation. PMID- 10815835 TI - Sequential labelling of microdamage in bone using chelating agents. AB - Basic fuchsin labels microcracks, but a series of stains is required to differentiate between preexisting and test-induced microcracks and to label their growth in vitro. Basic fuchsin and five chelating agents-alizarin complexone, calcein, calcein blue, oxytetracycline, and xylenol orange-were randomly assigned to label microcracks in sequential rib sections from 10 donors. The density, length, and location of the microcracks did not differ significantly between the six stains, suggesting that each was equally effective in detecting microcracks. Paired specimens of trabecular bone were machined from bovine tibiae, stained with oxytetracycline, and fatigued in compression. One specimen from each pair was then stained with xylenol orange. Preexisting microdamage was stained with oxytetracycline, propagating microcracks with both stains and new, test-initiated damage with xylenol orange. Chelating agents are site-specific markers of the initiation and growth of microcracks. PMID- 10815836 TI - Tibial plateau fracture as a measure of early estrogen-dependent bone fragility in rats. AB - The goals of this study were to develop a protocol to induce a compressive fracture at the tibial plateau of the rat knee in vitro and to determine if the biomechanical parameters provided a sensitive assessment of the early skeletal changes induced by estrogen deficiency. Sixty-one rats underwent an ovariectomy (n = 36) or sham operation (n = 25) and were maintained for 50 days after the procedure. Just before death, the proximal tibia of each animal was scanned with high-resolution x-ray tomography. From the three-dimensional images, the mean trabecular bone volume, thickness, and separation and the number of trabeculae were calculated. The knees were then harvested and mounted into a servohydraulic materials testing system so that the distal femoral condyle could be forced into the proximal tibial plateau until fracture. The fracture load of the ovariectomized rats was 24% less than that of the rats that had the sham operation. Similarly, the structural stiffness of the ovariectomized knees was decreased by 22%. Both of these differences were statistically significant (p < 0.01) and were explained by differences in trabecular bone volume (r = 0.56, p < 0.0001 and r = 0.42, p < 0.005, respectively). The other measures of trabecular bone structure were correlated with the volume and did not improve the prediction by the biomechanical parameters. These data demonstrate that biomechanical testing of the tibial plateau in rats can quantify the structural consequences of estrogen deficiency at an early time point before they become apparent at other bone sites, such as the lumbar spine. PMID- 10815837 TI - Vertebral trabecular bone microscopic tissue elastic modulus and hardness do not change in ovariectomized rats. AB - Ovariectomized rats have been used extensively and have received substantial acceptance as animal models for postmenopausal osteoporosis. However, little is known about the microscopic tissue properties of rat vertebral bone, especially during osteoporosis caused by estrogen depletion. This study applied a new nanoindentation technique to quantify the microscopic mechanical properties of vertebral trabecular bone tissue in ovariectomized rats. Seventeen-week-old Sprague-Dawley rats underwent an ovariectomy. After death at 37 weeks, the fraction of the trabecular bone area of the lumbar vertebrae (L4) was measured with scanning electron microscopy and the elastic modulus and hardness were determined with the nanoindentation technique. The bone area fraction was reduced significantly after the ovariectomy. However, the elastic modulus and hardness did not change significantly at the microscopic level. The results indicate that estrogen-dependent osteoporosis in rats manifests in a loss of bone mass whereas the elastic and hardness properties of the surviving bone tissue remain relatively unchanged. PMID- 10815838 TI - Nonlinear finite element modeling to evaluate the failure load of the proximal femur. PMID- 10815839 TI - Oligodendroglial cell behaviour in traumatic oedematous human cerebral cortex: a light and electron microscopic study. AB - Cortical biopsies of 12 patients with traumatic brain injuries have been used in the present study to examine oligodendroglial cell changes and reactivity. The samples were processed for light and transmission electron microscopy. Four main types of oligodendrocyte populations have been found: resting or unreactive oligodendrocytes, reactive oligodendrocytes, anoxic-ischaemic oligodendroglial cells and hyperthrophic phagocytic oligodendrocytes. The unreactive or resting oligodendrocyte type exhibited a fusiform or elongated shape, a clear or dense band of scarce perikaryal cytoplasm and a nucleus with peripheral heterororomatin masses. Clear or dense reactive oligodendrocytes showed increased amount of perikaryal citoplasm, dilated endoplasmic reticulum and nuclear envelope, numerous clear, oedematous mitochondria and dense bodies. These oligodendrocytes appeared associated with degenerated myelinated axons. Anoxic-ischaenmic oligodendrocytes showed lacunar enlargement of endoplasmic reticulum, dilated Golgi complex and enlargement and disassembly of nuclear envelope. They appeared also in contact with degenerated myelinated axons. Hypertrophic phagocytic oligodendrocytes were observed engulfing the associated degenerated myelinated axons, invading the myelin sheath, separating the myelin lamellae and exerting myelinolitic effects. Oligodendroglialpseudopodic expansions were observed phagocyting the axoplasmic matrix and leaving a huge vacuolar axoplasmic space. The vasogenic and cytotoxic components of traumatic brain oedema are discussed in relation with the oligodendroglial cell changes and reactivity. PMID- 10815840 TI - Early indication of emergence from vegetative state derived from assessments with the SMART--a preliminary report. AB - An explanatory analysis of data from serial assessments of 30 patients with a diagnosis of vegetative state (persistent vegetative state) was carried out. The data were gathered using the Sensory Modality Assessment and Rehabilitation Technique (SMART) and the Western Neuro Sensory Stimulation Profile (WNSSP) on the same day in the week post-admission and, thereafter, at 2 monthly intervals. Seven patients emerged from vegetative state during the study, a further six emerged later (by time of the analysis) and the rest remained in vegetative state. Recovery scores were calculated for each measure by subtracting the total score at any one assessment from the total score for the subsequent assessment; the largest recovery scores, shown by each patient, were noted. It was found that those who emerged later could be differentiated mathematically from those who did not emerge, using largest recovery score data from the SMART. This research supports previous PMID- 10815841 TI - Self-awareness of traumatically brain injured patients in the acute inpatient rehabilitation setting. AB - The self-awareness of brain injured patients in an acute inpatient rehabilitation programme was measured using the Functional Self-Appraisal Scale (FSAS), which compares patient and staff ratings of patient performance on tasks relevant for acute rehabilitation and in a format that is easy to administer. There was a significant difference between patients' and staffs ratings near admission, consistent with previous findings in post-acute settings that brain injured patients tend to overestimate their abilities relative to other raters. Patient and staff ratings tended to converge at time of discharge, likely reflecting patients' improvements on rehabilitation tasks rather than increased self awareness. The average difference between patients' and staffs ratings did not correlate with neuropsychological functioning on admission, supporting evidence that self-awareness early in the recovery process is a unique construct. Future research is outlined, including investigating the relationship of early levels of self-awareness following TBI to functional outcome. PMID- 10815842 TI - Outcome after shunt implantation in severe head injury with post-traumatic hydrocephalus. AB - OBJECT: Post-traumatic hydrocephalus (PTH) is considered a frequent complication after severe head injury (HI). There is little known about outcome following shunt implantation. METHODS: A hospital-based retrospective cross-over study investigated the outcome of 48 patients after severe HI, who had undergone ventricular shunt implantation due to PTH (40 males, mean age at injury 36 years, mean duration from HI to shunt implantation 27 weeks). Telephone interviews with the patients or with caring family members by means of a detailed questionnaire were performed after a mean observation period of 3.3 years after shunt implantation. Outcome was categorized using the Glasgow Outcome Scale (GOS): I: 12 patients, II: 7, III: 16, IV: 9, V: 4 at follow-up. CONCLUSIONS: 52.1% had a clear-cut benefit from shunt implantation, whereas 47.9% had not. Post-traumatic seizures were observed in 31 of 48 patients. Other complications after shunt implantation occurred in 20) patients. Revision of shunt implantation was performed in 15 patients (nine due to technical failure, three haemorrhage, one delayed primary wound closure, and two unknown). Two patients clearly deteriorated after operation (one severe frontal bleeding, one sepsis). The best predictive parameter for outcome after shunt implantation was the pre-operative status, patients in a better clinical condition (pre-operative GOS score 3-severe disabled vs 4-persistent vegetative state) had a better outcome. Patient's age at injury did not seem to influence the outcome. Clinical and computertomographic findings were of rather moderate predictive value as regards short- and long-term outcome after shunt implantation. Cisternography does not seem to be of additional help in the establishment of definite diagnosis of PTH. PMID- 10815843 TI - Location of the traumatic subdural hygroma: role of gravity and cranial morphology. AB - Traumatic subdural hygroma (TSH) is frequently bilateral and locates on the top of the head in a supine position. It suggests that the gravity and cranial posture act a certain role. The authors tried to test this hypothesis. The computed tomographic (CT) scans or magnetic resonance (MR) images of 86 consecutive patients with TSH were re-evaluated. The symmetry of the cranium, the posture of the head during the radiological examinations, and the location of the lesion were all checked. The cranium was symmetrical in 47 patients and asymmetrical in 39 patients. TSH was more commonly bilateral in patients with symmetrical cranium than those with asymmetrical cranium (77% vs 62%). The asymmetrical cranium tended to turn to the flat side. It was more frequently oblique in MR images, which has a long scanning time, than in CT (29% vs 18%). In 39 asymmetric craniums, TSH was bilateral and it was symmetrical in 14 cases. In the remaining 25 cases, TSH located opposite to the flat side in 18 cases. In seven patients with the same side TSHs, four patients had it on the side of atrophy, two on the opposite side of a mass lesion. The gravity and cranial posture can predict the location of TSH. TSH usually occurs at the least pressure in the cranium as a lesion of ex vacuo. PMID- 10815844 TI - Disability pensions in relation to traumatic brain injury: a population study. AB - From a Danish national register of hospitalizations, all patients were identified who had a discharge diagnosis of traumatic brain injury between the years 1979 1993 inclusive, at ages 18-66 years inclusive. These were classified as having suffered either a concussion (n = 74,398), a cranial fracture (n = 4,452) or a cerebral contusion (n = 8,141). Patients in each of these groups were then checked in annual registers of disability pension awards between 1979-1995. Disability pensions had been awarded to 16% of the concussion group, 18% of the fracture group, and 33% of the contusion group. Date of application, grounds for the application, and the pension level awarded were noted. Analysis of the date of application for the disability pension revealed that in all groups a high proportion of the pension applications had been made prior to the injury. Among the concussion group, the pension award appeared to be independent of the injury itself. Rather, being awarded a disability pension appeared to be related to conditions which themselves are risk factors for a traumatic brain injury, e.g. chronic skeletomuscular disease and psychiatric disorders including alcoholism. Comparison with population statistics revealed that the relative risk of being assigned a low or intermediate disability pension is markedly elevated among the concussion group, especially at younger ages and among males. For the fracture and contusion groups, a clear post-injury elevation in rate was detectable, especially for highest level pensions. PMID- 10815845 TI - Ambulatory care for traumatic brain injuries in the US, 1995-1997. AB - The burden of mild traumatic brain injury (TBI) is not well understood at the national level, but hospitalization rates show a decline over time. This paper describes ambulatory care for TBI patients at physician offices, hospital outpatient departments, and emergency departments (EDs) in comparison with non TBI visits for the US during 1995-1997. An estimated 1.4 million visits for TBI were made each year for an average annual rate of 5.4/1,000 population. A decline in annual visit rate was noted during 1995-1997. Visit rates were higher for those aged 0 -14 and 75 and older. Falls (44%) and motor vehicles (28%) were the primary injury causes. Rural-urban differences were found, also in comparison with non-TBI. In 23% of visits to EDs, a CT scan was ordered or performed and in 33%, a mental status exam was conducted. Further investigations are warranted to describe ambulatory care for TBI in more detail, particularly in light of a decline in hospitalization rates. PMID- 10815846 TI - Cocaine-associated stroke: three cases and rehabilitation considerations. AB - Cocaine-associated stroke (CAS) is an important cause of disability, especially among younger adults. Improved management has increased survival but little has been discussed about rehabilitation, including medication management. Therefore, experience and therapeutic drug management are described during inpatient rehabilitation with three patients with CAS. Case 14 is a 50-year-old male with a history of hypertension who presented with right hemiparesis, aphasia and depression. He was treated with paroxetine for depression and bromocriptine for poor initiation with a good response, improving by 50 FIM points in 44 days. Case 2 is a 44 year-old female with quadriparesis, aphasia, and deficits in attention and initiation. Methylphenidate for attention deficits and bromocriptine for poor initiation was associated with an excellent functional gain (50 FIM points in 37 days). She eventually returned to work. Case 3 is a 46-year-old female with a history of hypertension who presented with right hemiparesis, aphasia and depression. Without neuropharmacologic intervention, she gained 35 FIM points during an uneventful 47 day rehabilitation stay. Acutely, cocaine can induce cerebral vasoconstriction, cerebrovascular spasm, cerebral vasculitis and intracerebral haemorrhage. Chronic use depletes and destroys dopaminergic pathways, which may be a major factor in depression, and attention and initiation deficits-all observed in these cases. Generally, rapid improvements were seen in mood and cognition in two cases where medication was used. Based on the current literature and pathophysiology of CAS, it is suggested that trials of dopaminergic agents for cognition and extremely cautious use of buproprion for depression may be warrented. Details of the above cases and the practical and theoretical issues of neuropharmacologic intervention in CAS are discussed. PMID- 10815847 TI - [Fatigue of the respiratory muscles due to maximal exercise on 2 different ergometers]. AB - This study examines relationships between physical exercise and fatigue of ventilatory muscles according to training, intensity of exercise, and the ergometer. Twenty-two athletes were submitted to maximal triangular exercise on treadmill (TR) and bicycle (BE). Maximal inspiratory (PImax) and expiratory pressures and a spirometry were measured before and after exercise. In the "high level physical capacity" group (GR1) and "medium level physical capacity" group (GR2), PImax (128.4 +/- 36.8 cmH2O) and PEmax (175.7 +/- 63.8 cmH2O) after exercise on TR decreased significantly (140.0 +/- 32.6, p < .05; at rest: 196.8 +/- 59.8 cmH2O, p < .01). After exercise on BE, PImax and PEmax in GR1 and GR2 were significantly smaller than at rest (p < .001). In GR1, forced vital capacity (FVC) was significantly decreased after exercise on TR (5.22 +/- 0.8 vs. 4.99 +/- 0.6 l; p < .05). PMID- 10815848 TI - Ventilatory threshold and maximal oxygen uptake in present triathletes. AB - The aim of this study was to determine the physiological profile of young triathletes who began triathlon competition as their first sport. Twenty-nine male competitive triathletes (23 regionally and nationally ranked triathletes and 6 elite, internationally ranked triathletes) performed two tests, one on a cycle ergometer (CE VO2max) and one on a treadmill (TM VO2max). Results showed (a) no difference between CE VO2max and TM VO2max in the triathletes (69.1 +/- 7.2 vs. 70.2 +/- 6.2 mL x kg(-1) x min(-1), respectively), (b) values of CE VO2max and TM VO2max in elite triathletes (75.9 +/- 5.2 and 78.5 +/- 3.6 mL x kg(-1) x min(-1), respectively) that were comparable to those reported in elite single-sport athletes in these specialities, and (c) although the ventilatory threshold (Th(vent)) was similar in CE and TM, TM Th(vent) was consistently lower for triathletes than TM Th(vent) usually reported for runners. PMID- 10815849 TI - Health implications of musculoskeletal fitness. AB - Although it is well documented that physical activity participation can bring about improvements in musculoskeletal fitness, related health implications have only been substantiated in the elderly. Currently, interpretations of the results of musculoskeletal fitness appraisals for the general population are based largely on an intuitive belief that enhanced musculoskeletal fitness is associated with higher levels of health throughout adulthood. Musculoskeletal fitness was measured in 312 females and 259 males aged 15-69 years. Health levels were determined using two previously validated questionnaires and expressed as composite health scores derived from principal components analysis. Grip strength, push strength, pull strength, push-ups, and trunk forward flexion were significant predictors of health scores after controlling for the potentially confounding effects of gender age, aerobic fitness, waist circumference, and smoking status. These findings indicate that musculoskeletal fitness is related to health in males and females aged 15 to 69 years. PMID- 10815850 TI - Effects of exercise and alkalosis on serum insulin-like growth factor I and IGF binding protein-3. AB - This investigation examines the effects of orally induced alkalosis on serum IGF I and IGFBP3 concentrations in response to an acute 90-s bout of high intensity cycle exercise. Ten healthy, active men, ages 24.60 +/- 4.90 years, participated in a randomized, double-blind, counterbalanced trial order with a cross-over design. Subjects ingested an experimental bicarbonate solution or a placebo solution. Blood was sampled at baseline; pre-exercise; and 0, 5, 10, and 30 min postexercise. The pH between groups for pre-exercise and postexercise time points differed significantly (p < or = .05) in the experimental condition (from 7.42 +/ 0.01 to 7.35 +/- 0.02) versus the placebo condition (from 7.36 +/- 0.01 to 7.25 +/- 0.03). Increases in IGF-I over resting conditions occurred with placebo conditions at 5 and 10 min postexercise and in the experimental condition at 5 min postexercise. Concentrations of IGFBP3 were elevated above baseline at IP in both experimental and placebo conditions. PMID- 10815852 TI - Congenital generalized myofibromatosis in a neonate. PMID- 10815851 TI - Acitretin for hypertrophic lichen planus-like reaction in a burn scar. PMID- 10815853 TI - Read the Lancet, or else! PMID- 10815854 TI - Troglitazone improves psoriasis and normalizes models of proliferative skin disease: ligands for peroxisome proliferator-activated receptor-gamma inhibit keratinocyte proliferation. AB - BACKGROUND: Psoriasis is often treated with agents that activate nuclear hormone receptors for glucocorticoids, retinoids, and vitamin D. The peroxisome proliferator-activated receptor-gamma (PPARgamma) is a related nuclear hormone receptor that can be activated by its ligands, including the thiazolidinediones. OBJECTIVE: To assess whether treatment with troglitazone, a currently available thiazolidinedione used to treat diabetes mellitus, has an effect on psoriasis in normoglycemic patients and whether ligands for PPARgamma have an effect on models of psoriasis. DESIGN: Open-label administration of troglitazone in patients with psoriasis and evaluation of drug actions in cellular, organ, and transplant models of psoriasis. SETTING: University and community hospital outpatient departments and university laboratories. PATIENTS: Patients with chronic, stable plaque psoriasis and control subjects. Five patients with psoriasis received troglitazone (none withdrew); 10 different untreated patients and 10 controls provided tissue samples. INTERVENTIONS: Oral troglitazone therapy at various dosages in patients with psoriasis; also, use of troglitazone, ciglitazone, and 15-deoxy-delta-12,14-prostaglandinJ2 in psoriasis models. MAIN OUTCOME MEASURES: Investigator-determined clinical results in patients and cell counts and histological evidence in models. RESULTS: All patients' psoriasis improved substantially during troglitazone therapy. Peroxisome proliferator-activated receptor-gamma was expressed in human keratinocytes; ligands for PPARgamma inhibited the proliferation of normal and psoriatic human keratinocytes in culture. Troglitazone treatment normalized the histological features of psoriatic skin in organ culture and reduced the epidermal hyperplasia of psoriasis in the severe combined immunodeficient mouse and human skin transplant model of psoriasis (P<.05 compared with untreated controls). CONCLUSIONS: Peroxisome proliferator-activated receptor-gamma might be a useful intracellular target for the treatment of psoriasis; further study is needed to assess the clinical value of ligands for PPARgamma, including troglitazone. PMID- 10815855 TI - 308-nm excimer laser for the treatment of psoriasis: a dose-response study. AB - OBJECTIVE: To determine the dose-response relationship of excimer laser-generated 308-nm UV-B radiation for treating psoriasis. DESIGN: Pilot study with a before after design. SETTING: A university dermatology service. PATIENTS: Thirteen consecutive patients with at least 4 large, stable psoriasis plaques. INTERVENTIONS: Excimer laser-generated 308-nm UV-B radiation was given to each of 4 plaques, which received 1, 2, 4, and 20 treatments, respectively. Untreated areas within each plaque served as controls. Within each plaque, 8 doses based on multiples of a predetermined minimal erythema dose (MED) were tested in distinct sites. The multiples were 0.5 and 1 (low dose); 2, 3, 4, and 6 (medium dose); and 8 and 16 (high dose). At every treatment, the dose for each site remained fixed at the same MED multiple. A psoriasis severity index score was determined for each area before, every 2 weeks during, and 2 and 4 months after treatment. RESULTS: The mean+/-SD MED was 203.03+/-57.84 mJ/ cm2. Treatment with high fluences produced significantly better results than that with medium and low fluences at weeks 4, 6, 8, and 10 (P<.05). At 4 months' follow-up, all sites that received low or medium fluences had recurrences, whereas those that underwent a single treatment at 8 and 16 MED multiples remained in remission. CONCLUSIONS: With 308-nm UV-B radiation generated by an excimer laser, it is possible to clear psoriasis with as little as 1 treatment with moderately long remission. In contrast to traditional phototherapy techniques, this handheld excimer laser UV-B therapy is selectively directed toward lesional skin, thus sparing the surrounding normal skin from unnecessary radiation exposure. Treatment of other inflammatory diseases and limited psoriasis seems reasonable to pursue with this modality. PMID- 10815856 TI - Flashlamp-pumped pulsed dye laser for hemangiomas in infancy: treatment of superficial vs mixed hemangiomas. AB - OBJECTIVE: To study in a compared manner the efficacy of flashlamp-pumped pulsed dye laser (FPDL) therapy for superficial and mixed hemangiomas. DESIGN: Nonrandomized control trial. SETTING: Department of Lasermedicine, General Hospital Neukolln, Berlin, Germany. PATIENTS: To investigate variation in response to treatment, a prospective study of 165 children with 225 separate hemangiomas treated with the FPDL was undertaken. Patients were aged 2 days to 7 years; mean follow-up was 5 months. INTERVENTIONS: During a 2 1/2-year period, we administered 332 treatments, for a mean+/-SD of 2.0+/-1.1 treatments per patient. MAIN OUTCOME MEASURE: Patients received therapy until the lesion was almost clear or until the lesion did not respond to treatment. Evaluation was performed by comparing pretreatment and posttreatment photographs. In addition, pathologic flow of vessels and thickness were determined before, during, and after completion of therapy with color-coded duplex sonography. RESULTS: In the first group of 100 patients with 153 flat cutaneous hemangiomas, 52 hemangiomas (34%) had excellent results; 80 (52%) had good results; and 21 (14%) showed proliferation of the subcutaneous component, although these lesions were flat at first presentation. Of the 54 mixed hemangiomas, 33 (61%) had continued proliferation of the subcutaneous component. The cutaneous component responded to therapy in 21 hemangiomas (39%), whereas the subcutaneous component of the mixed hemangiomas remained unchanged. No lesions in this group involuted completely, and therapy was discontinued because of relatively poor response. Twelve (67%) of 18 patients with superficial hemangiomas in the involution phase had excellent results and 6 (33%) had good results. CONCLUSIONS: Treatment with the FPDL is effective and may be the treatment of choice for superficial cutaneous hemangiomas at sites of potential functional impairment and on the face. Hemangiomas with a deep component do not benefit from FPDL treatment because the efficacy of the FPDL is limited by its depth of vascular injury. Furthermore, early therapeutic intervention with the FPDL may not prevent proliferative growth of the deeper or subcutaneous component of the hemangioma despite early intervention. PMID- 10815857 TI - Personal and clinical skin cancer prevention practices of US women physicians. AB - OBJECTIVE: To document physician clinical and personal skin cancer prevention practices and associated characteristics. DESIGN: A cross-sectional questionnaire survey of a representative sample of US women physicians. SETTING: Mail survey. SUBJECTS: Three thousand thirty-two nondermatologists and 95 dermatologists. MAIN OUTCOME MEASURE: Personal and clinical practices. RESULTS: Twenty-seven percent of nondermatologists counseled or screened their typical patients on skin cancer or sunscreen use at least once a year, while 49% did so less frequently, and 24% never counseled or screened at all. Of the 95 dermatologists, two thirds reported counseling or screening their typical patients at every visit. In bivariate analysis of nondermatologists, the distribution of counseling or screening was significantly (P<.05) associated with the following personal and professional characteristics: frequent sunscreen use, recent (within 2 years) skin examination, good health status, a primary care specialty, self-confidence in counseling or screening, extensive training in counseling or screening, high perceived relevance to the practice of the counseling or screening, nonurban practice site, and nonhospital-based or non-medical school-based practice. We found that 48% of all physicians always or nearly always used sunscreen, and 25% had received a clinical skin examination in the previous 2 years. CONCLUSIONS: Although many primary care physicians report ever counseling or screening their typical patients about skin cancer and sunscreen use, increased professional education for primary care physicians could improve patient counseling about skin cancer prevention. PMID- 10815858 TI - Subsequent cancers after in situ and invasive squamous cell carcinoma of the skin. AB - OBJECTIVES: To compare cancer risks after in situ and invasive squamous cell carcinoma (SCC) of the skin and to determine whether these 2 forms of cancer differ in prognostic significance. PATIENTS: Subsequent events after in situ and invasive SCC were studied in the Swedish Family-Cancer Database, in which cancer data were obtained from the Swedish Cancer Registry from 1958 to 1996. Among 22293 patients with in situ SCC, 3940 had first invasive cancer; among 17637 patients with invasive SCC, 3624 had a second occurrence of cancer. MAIN OUTCOME MEASURE: Standardized incidence ratios (SIRs), ratios of the observed to expected number of cases, served as a measure of relative risk. For overall risks, cases diagnosed within the first year of follow-up were omitted. RESULTS: The median age of onset was 72 to 73 years for in situ and invasive SCC, respectively. Standardized incidence ratios of all cancers were increased after in situ SCC (men-women, 1.5:1.3) and invasive SCC (men-women, 1.9:1.5). The subsequent occurrences of cancer and their SIRs were similar after in situ and invasive SCC, with skin cancer showing the highest SIR of 6.4:10.0. Among discordant cancers, increased SIRs were recorded for melanoma and a group of malignant neoplasms observed in patients with immunosuppression, including lymphoma and oral cancers. Subsequent cancers in the salivary glands and nasal cavity also showed increased SIRs, particularly after invasive SCC. CONCLUSION: Risks of subsequent cancers, including skin cancer, melanoma, and internal cancers, showed similar patterns in patients with in situ and invasive SCC, suggesting that the 2 groups have a similar susceptibility to cancer. PMID- 10815859 TI - Lichenoid dermatitis in paraneoplastic pemphigus: a pathogenic trigger of epitope spreading? AB - BACKGROUND: In select cases, lichen planus has been observed to be a paraneoplastic condition sometimes associated with paraneoplastic pemphigus, a disease featuring autoantibodies directed against plakin proteins, desmogleins 3 and 1, and a still uncharacterized 170-kd antigen. Epitope spreading describes the phenomenon where underlying chronic inflammation leads to the sequential recognition of new epitopes on self-proteins over time. OBSERVATIONS: Five of 6 patients diagnosed as having paraneoplastic pemphigus had concomitant clinical and histological features of lichen planus. In 1 patient, results of the initial indirect immunofluorescence on rat bladder were negative and only 2 of the 5 antigens were identified by immunoprecipitation. After 1 year of worsening disease, repeated testing confirmed the presence of antibodies directed against all 6 of the implicated antigens, supportive of our hypothesis that epitope spreading may occur in paraneoplastic pemphigus. CONCLUSIONS: Lichenoid eruptions may predispose to an early evolutionary stage of paraneoplastic pemphigus. Cell mediated autoimmunity at the dermoepidermal junction may promote the exposure of self-antigens and the development of subsequent and progressive humoral autoimmunity. As such, paraneoplastic pemphigus may demonstrate epitope spreading in a human, humoral-mediated autoimmune disease. PMID- 10815860 TI - Sporadic trichoepithelioma demonstrates deletions at 9q22.3. AB - BACKGROUND: Trichoepithelioma (TE) is a benign cutaneous tumor that originates from hair follicles and occurs either in multiple or solitary lesions. Multiple TE is transmitted as an autosomal dominant trait, and a region at 9p21 is thought to be involved in the tumorigenesis. Solitary TE occurs more commonly than multiple TE and is not inherited. Histologically, TE tumors contain horn cysts and abortive hair papillae. A basal cell carcinoma appearance in some or all regions of a TE tumor can happen. In sporadic basal cell carcinoma, frequent deletions at 9q22.3 (Drosophila patched gene) have occurred. The objective of this study is to test whether loss of heterozygosity (LOH) on either 9p21 or on chromosome 9q22.3 could be detected in archival sporadic TE. OBSERVATIONS: We studied 29 randomly selected cases of sporadic TE by microdissection and polymerase chain reaction using paraffin-embedded, formalin-fixed tissue specimens on glass slides. Analysis was performed with the polymorphic markers IFNA and D9S171 (9p21) as well as D9S15, D9S303, D9S287, and D9S252 (9q22.3). RESULTS: The LOH at 9q22.3 was identified in 14 (48%) of 29 cases with at least 1 marker, while LOH could not be demonstrated using the markers IFNA and D9S171 (9p21). CONCLUSIONS: The results show that the Drosophila patched gene LOH can be frequently identified in paraffin-embedded sporadic TE after routine processing and indicates a common gatekeeper mechanism for both TE and basal cell carcinoma. PMID- 10815861 TI - Epitope spreading in paraneoplastic pemphigus: autoimmune induction in antibody mediated blistering skin diseases. PMID- 10815862 TI - Generalized erythematous plaques. Progressive symmetric erythrokeratodermia (PSEK) (erythrokeratodermia progressiva symmetrica). PMID- 10815863 TI - A persistent lower lip lesion. Verruciform xanthoma. PMID- 10815864 TI - Mucosal erosions and bullae in a child. Pemphigus vulgaris in a child with tinea capitis. PMID- 10815865 TI - A patient with dermatomyositis and linear streaks on the back. Centripetal flagellate erythema (CFE) associated with dermatomyositis. PMID- 10815866 TI - Simultaneous infection by human herpesvirus 7 and human parvovirus B19 in papular purpuric gloves-and-socks syndrome. PMID- 10815867 TI - My guardian angel wore a big hat at Sea World! PMID- 10815868 TI - Association of Sweet syndrome and erythema nodosum. PMID- 10815869 TI - The 21st century macule is not the Willan but the Plenck macule. PMID- 10815871 TI - Treatment of severe psoriasis with anti-CD25 monoclonal antibodies. PMID- 10815870 TI - Constitutive speckled vascular mottling of the skin resembling Bier white spots: lack of venoarteriolar reflex in dermal arterioles. PMID- 10815872 TI - Allogenic keratinocytes suspended in human fibrin glue used for wound healing support in chronic leg ulcers. PMID- 10815873 TI - Heavy naphthen oil exhibits antipsoriatic efficacy in vivo and antiproliferative as well as differentiation-inducing effects on keratinocytes in vitro. PMID- 10815874 TI - Improvement of atopic dermatitis after discontinuation of topical corticosteroid treatment. PMID- 10815875 TI - Antisense approaches enter the clinic. PMID- 10815876 TI - Angiogenesis and the multistage development of lung cancers. PMID- 10815877 TI - Discodermolide: just another microtubule-stabilizing agent? No! A lesson in synergy. PMID- 10815878 TI - Angiogenic squamous dysplasia in bronchi of individuals at high risk for lung cancer. AB - Lung carcinogenesis is assumed to be a multistep process, but detailed understanding of the sequential morphological and molecular changes preceding invasive lung cancer remains elusive. To better understand early lung carcinogenesis, we initiated a program of fluorescence bronchoscopy in smokers at high risk for lung cancer. In the bronchial biopsies from these subjects, we observed a unique lesion consisting of capillary blood vessels closely juxtaposed to and projecting into metaplastic or dysplastic squamous bronchial epithelium, angiogenic squamous dysplasia (ASD). Serial sections of the capillary projections confirmed that they represent intramucosal capillary loops. Microvessel density in ASD was elevated in comparison to normal mucosa (P = 0.0003) but not in comparison to other forms of hyperplasia or dysplasia. ASD thus represents a qualitatively distinct form of angiogenesis in which there is architectural rearrangement of the capillary microvasculature. Genetic analysis of surface epithelium in a random subset of lesions revealed loss of heterozygosity at chromosome 3p in 53% of ASD lesions. No confirmed p53 mutations were identified. Compared with normal epithelium, proliferative activity was markedly elevated in ASD lesions. ASD occurred in 54 of 158 (34%) high-risk smokers without carcinoma and in 6 of 10 patients with squamous carcinoma who underwent fluorescence bronchoscopy. One early-stage invasive carcinoma was noteworthy for the occurrence of ASD juxtaposed to invasive tumor. Seventy-seven (59%) of the ASD lesions were detected by abnormal fluorescence alone. Twenty bronchial sites (11 patients) were rebiopsied 1 year after the initial diagnosis. At nine (45%) of these sites, the lesion was found to persist. The lesion was not present in biopsies from 16 normal nonsmoker control subjects. The presence of this lesion in high-risk smokers suggests that aberrant patterns of microvascularization may occur at an early stage of bronchial carcinogenesis. PMID- 10815879 TI - A phase I trial of c-Raf kinase antisense oligonucleotide ISIS 5132 administered as a continuous intravenous infusion in patients with advanced cancer. AB - Raf proteins play a central role in the mitogen-activated protein kinase signaling pathway and hence are involved in oncogenic transformation and tumor cell proliferation. ISIS 5132 is a 20-base antisense phosphorothioate oligodeoxyribonucleotide that specifically down-regulates c-raf expression. We report here an initial study of the safety and tolerability of an i.v. infusion of ISIS 5132 in patients with advanced cancer. A continuous i.v. infusion of ISIS 5132 was administered for 21 days every 4 weeks to 34 patients with a variety of solid tumors refractory to standard therapy. The dose of ISIS 5132 was increased in sequential cohorts of patients, as toxicity allowed, until a final dose of 5.0 mg/kg body weight was reached. Toxicity was scored by common toxicity criteria, and tumor response was monitored. Pharmacokinetic studies were performed for 30 patients treated at doses of < or =4.0 mg/kg/day. The initial dose of ISIS 5132 was 0.5 mg/kg body weight and was successfully increased incrementally to 5.0 mg/kg body weight. Toxicities through the 4.0 mg/kg dose level were not dose limiting. Side effects were minimal and could not be specifically related to ISIS 5132. Two patients had prolonged stabilization of their disease, and one patient with ovarian carcinoma had a significant response with a 97% reduction in CA-125 levels. ISIS 5132, an antisense oligonucleotide against c-raf, was well tolerated at doses up to and including 4.0 mg/kg/day by 21-day continuous i.v. infusion and demonstrated antitumor activity at the doses tested. PMID- 10815880 TI - A phase I trial of a recombinant vaccinia virus expressing prostate-specific antigen in advanced prostate cancer. AB - A recombinant vaccinia virus encoding human prostate-specific antigen (rV-PSA) was administered as three consecutive monthly doses to 33 men with rising PSA levels after radical prostatectomy, radiation therapy, both, or metastatic disease at presentation. Dose levels were 2.65 x 10(6), 2.65 x 10(7), and 2.65 x 10(8) plaque forming units. Ten patients who received the highest dose also received 250 microg/m2 granulocyte-macrophage colony-stimulating factor (GM-CSF) as an immunostimulatory adjunct. No patient experienced any virus-related effects beyond grade I cutaneous toxicity. Pustule formation and/or erythema occurred after the first dose in all 27 men who received > or =2.65 x 10(7) plaque forming units. GM-CSF administration was associated with fevers and myalgias of grade 2 or lower in 9 of 10 patients. PSA levels in 14 of 33 men treated with rV-PSA with or without GM-CSF were stable for at least 6 months after primary immunization. Nine patients remained stable for 11-25 months; six of these remain progression free with stable PSA levels. Immunological studies demonstrated a specific T-cell response to PSA-3, a 9-mer peptide derived from PSA. rV-PSA is safe and can elicit clinical and immune responses, and certain patients remain without evidence of clinical progression for up to 21 months or longer. PMID- 10815881 TI - Correlation of genetic instability with mismatch repair protein expression and p53 mutations in non-small cell lung cancer. AB - To examine the etiological association of genetic instability in lung tumorigenesis, we investigated the frequency of microsatellite instability (MI) of eight dinucleotide repeat markers in 68 patients with non-small cell lung cancer. Twenty-eight patients (41.2%) evidenced instability in multiple tested microsatellite markers ranging from 3-7 and were defined as MI-positive patients. MI occurred more frequently in patients suffering from squamous cell lung carcinoma (P = 0.004). We examined the association between MI and expression of hMLH1 mismatch repair protein by immunohistochemical analysis of hMLH1 protein in paraffin-embedded tumors from 64 patients. Twenty MI-positive patients (76.9%) had no expression of hMLH1 protein. The data showed that MI was associated with altered hMLH1 expression (P = 0.03). To examine the role of genetic instability in the previous identified small intragenic deletion of the p53 gene, we explored the association between MI and p53 gene mutations. All patients, except one, containing small intragenic deletion in p53 gene showed MI (P = 0.018). In addition, we found that MI was not associated with the prognosis. Our data suggest that MI plays a significant role in non-small cell lung cancer tumorigenesis in Taiwan and that MI is associated with the altered expression of hMLH1 mismatch repair protein. In addition, MI may be involved in frequent small intragenic deletions of p53 gene. PMID- 10815882 TI - Mouse macrophage metalloelastase gene transfer into a murine melanoma suppresses primary tumor growth by halting angiogenesis. AB - Mouse macrophage metalloelastase (MME) has been associated with the generation of angiostatin, an internal fragment of plasminogen, which inhibits angiogenesis. To clarify whether tumor cells that consistently generate MME can suppress angiogenesis and, therefore, inhibit the growth of primary tumors in vivo, we transfected a cDNA coding for MME into murine B16-BL6 melanoma cells that grow rapidly and are MME deficient. The generation of active MME in MME-transfected clones was confirmed by immunoprecipitation followed by in vitro cleavage of plasminogen. Subcutaneous implantation of these stable clones in C57BL/6 mice inhibited primary tumor growth by an average of 73% (P = 0.00002), which directly correlated with a significant reduction of blood vessel formation (approximately 76%) in such tumors. Microangiography revealed massive angiogenesis in control tumors (mock and vector); however, in MME-transfected primary tumors it demonstrated a decreased and disrupted vascular network. Western blot analysis using a specific anti-mouse angiostatin antibody demonstrated a strong 38-kDa immunoreactive band in MME-transfected tumors and in the serum of mice bearing those tumor cells. These results show that placing MME gene directly into B16-BL6 melanoma cells is an effective approach to suppress primary tumor growth in vivo because it halts angiogenesis. Our data provide a feasible and promising strategy for gene therapy of cancer by targeting tumor vasculature. PMID- 10815883 TI - Antisense TRPM-2 oligodeoxynucleotides chemosensitize human androgen-independent PC-3 prostate cancer cells both in vitro and in vivo. AB - Although numerous chemotherapeutic regimens have been evaluated for patients with hormone-refractory prostate cancer, none has improved survival. Testosterone repressed prostate message-2 (TRPM-2), which is highly up-regulated after androgen withdrawal and during androgen-independent progression in prostate cancer, has been shown to inhibit apoptosis induced by various kinds of stimuli. The objectives in this study were to test whether antisense (AS) oligodeoxynucleotides (ODNs) targeted against TRPM-2 enhance chemosensitivity in human androgen-independent prostate cancer PC-3 cells both in vitro and in vivo. Initially, the potency of 10 AS ODNs targeting various regions of the TRPM-2 mRNA were evaluated, and the AS ODN targeted to the TRPM-2 translation initiation site (AS ODN#2) was found to be the most potent sequence for inhibiting TRPM-2 expression in PC-3 cells. Despite significant dose-dependent and sequence specific suppression of TRPM-2 expression, AS ODN#2 had no effect on growth of PC 3 cells both in vitro and in vivo. However, pretreatment of PC-3 cells with AS ODN#2 significantly enhanced chemosensitivity of Taxol (paclitaxel) and mitoxantrone in vitro. Characteristic apoptotic DNA laddering and cleavage of poly(ADP-ribose) polymerase were observed after combined treatment with AS ODN#2 plus paclitaxel or mitoxantrone but not with either agent alone. In vivo administration of AS ODN#2 plus either paclitaxel or mitoxantrone significantly decreased PC-3 tumor volume by 80 or 60%, respectively, compared with mismatch control ODN plus either paclitaxel or mitoxantrone. In addition, terminal deoxynucleotidyl transferase-mediated nick end labeling staining revealed increased apoptotic cells in tumors treated with AS ODN#2 plus paclitaxel or mitoxantrone. These findings confirm that TRPM-2 overexpression confers resistance to cytotoxic chemotherapy in prostate cancer cells and illustrates the potential utility of combined treatment with AS TRPM-2 ODN plus chemotherapeutic agents for patients with hormone-refractory prostate cancer. PMID- 10815884 TI - Methylation of the neutral endopeptidase gene promoter in human prostate cancers. AB - Neutral endopeptidase 24.11 (NEP) is a cell surface peptidase expressed by prostatic epithelial cells that cleaves and inactivates neuropeptide growth factors implicated in the growth of androgen-independent prostate cancer (PC). Decreased NEP expression in hormone-refractory metastatic PCs can result from hormonal therapies because NEP transcription is induced by androgens and down regulated by androgen withdrawal. NEP is encoded by a gene that contains a 5' CpG island spanning a transcriptional regulatory region. In this study, we investigate whether DNA hypermethylation of the NEP promoter accompanies decreased NEP expression in PC cell lines and whether it occurs in human PC tissues in vivo. DNA isolated from PC cell lines and from normal and neoplastic human prostate tissues was restriction-digested with a methylation-sensitive restriction endonuclease and analyzed by Southern blot using a 5' sequence specific NEP probe. Methylation-specific PCR was performed using PCR primers designed to discriminate between methylated and unmethylated alleles, and reverse transcription-PCR using NEP-specific primers was performed on cDNA extracted from PC cells treated with 5-aza-2'-deoxycytidine. Methylation of the NEP promoter was present in androgen-independent PC cell lines but not in androgen-dependent or small-cell derived PC cell lines and in 3 of 21 (14%) primary PCs from patients with androgen-dependent disease. Exposure of PC cells to the demethylating agent 5-aza-2'-deoxycytidine led to an increase in NEP transcripts in DU-145 and PC-3 cells. These data show that hypermethylation of the 5' CpG NEP island is associated with a loss of NEP expression in PC. Loss of NEP expression via hypermethylation of the NEP promoter may contribute to the development of neuropeptide-stimulated PCs. PMID- 10815885 TI - Restoration of Th1 cytokine synthesis by T cells of patients with chronic myelogenous leukemia in cytogenetic and hematologic remission with interferon alpha. AB - Chronic myelogenous leukemia (CML) is a disorder of the hematopoietic stem cell that results in malignant expansion of myeloid cells with a cytogenetic abnormality, the translocation between chromosomes 9 and 22 known as the Philadelphia chromosome. Treatment with IFN-alpha has proven to be an effective therapy, inducing cytogenetic remission in CML patients. However, it is unknown whether IFN-alpha can restore normal immune function for patients who achieve a complete cytogenetic remission. To address this question, we used a method of intracellular staining and flow cytometric analysis to ascribe the syntheses of Th1 or Th2 cytokines to T-cell subsets of patients in chronic, in accelerated, and in blast crisis phases as well as patients who had achieved a complete cytogenetic remission with IFN-alpha. We assessed the cytoplasmic synthesis of cytokine in phorbol ester (phorbol 12-myristate 13-acetate)-activated CD4+ and CD8+ T-cell subsets of 81 patients with various stages of CML and 21 normal controls. The percentages of CD4+ and CD8+ T cells from patients in chronic, in accelerated, and in blast crisis phases that synthesized Th1 cytokines interleukin (IL)-2, IFN-gamma, and tumor necrosis factor-alpha were significantly lower than those of remission patients and normal controls. Conversely, the percentages of CD4+ and CD8+ T cells of patients in chronic, in accelerated, and in blast crisis phases of CML preferentially synthesized the Th2 cytokine IL-10. Patients who achieved a durable complete cytogenetic remission for >2 years without maintenance IFN-alpha therapy restored their preference for a Th1 cytokine profile that is necessary for efficient cytotoxic T-cell function. PMID- 10815886 TI - Phase I trial of twice-weekly intravenous interleukin 12 in patients with metastatic renal cell cancer or malignant melanoma: ability to maintain IFN-gamma induction is associated with clinical response. AB - The aim of this study was to examine the tolerability, antitumor activity, and biological effects of a new schedule of i.v. recombinant human interleukin 12 (rhIL-12). Twenty-eight patients were enrolled in a Phase I trial in which rhIL 12 was administered twice weekly as an i.v. bolus for 6 weeks. Stable or responding patients were eligible to receive additional 6-week cycles until there was no evidence of disease or until tumor progression. Patient cohorts were treated with escalating doses of rhIL-12 (30-700 ng/kg). The maximum tolerated dose (MTD) was 500 ng/kg, with dose-limiting toxicities consisting of elevated hepatic transaminases and cytopenias. At the MTD (n = 14), there was one partial response occurring after 6 cycles of rhIL-12 in a patient with renal cell cancer. Two additional renal cell cancer patients treated at the MTD had prolonged disease stabilization, with one of these exhibiting tumor regression after 8 cycles of rhIL-12. IFN-gamma, IL-15, and IL-18 were induced in patients treated with rhIL-12. Whereas IFN-gamma and IL-15 induction were attenuated midway through the first cycle in patients with disease progression, those patients with tumor regression or prolonged disease stabilization were able to maintain IFN gamma, IL-15, and IL-18 induction. The down-modulation of IFN-gamma induction during rhIL-12 treatment did not relate to IL-10 production or alterations in rhIL-12 bioavailability but was associated with an acquired defect in lymphocyte IFN-gamma production in response to IL-12, IL-2, or IL-15. This defect could be partially overcome in vitro through combined stimulation with IL-12 plus IL-2. These findings show that the chronic administration of twice-weekly i.v. rhIL-12 is well-tolerated, stimulates the production of IL-12 costimulatory cytokines and IFN-gamma, and can induce delayed tumor regression. Strategies aimed at maintaining IFN-gamma induction, such as the addition of IL-2, may further augment the response rate to this schedule of rhIL-12. PMID- 10815887 TI - Vaccination of high-risk breast cancer patients with mucin-1 (MUC1) keyhole limpet hemocyanin conjugate plus QS-21. AB - Our objective was to determine whether an immune response can be generated against MUC1 peptide and against tumor cell MUC1 after vaccination with MUC1 keyhole limpet hemocyanin (KLH) conjugate plus QS-21 in breast cancer patients. Nine patients with a history of breast cancer but without evidence of disease were treated with MUC1-KLH conjugate plus QS-21, containing 100 microg of MUC1 and 100 microg of QS-21. s.c. vaccinations were administered at weeks 1, 2, 3, 7, and 19. Peripheral blood was drawn at frequent intervals to assess antibody titers. Skin tests were placed at weeks 1, 3, 9, and 21 to determine delayed type hypersensitivity reactions. Common toxicities included a local skin reaction at the site of the vaccine, usually of 4-5 days' duration, and mild flu-like symptoms usually of 1-2 days' duration. High IgM and IgG antibody titers against synthetic MUC1 were detected. IgG antibody titers remain elevated from a minimum of 106-137 weeks after the first vaccination. Binding of IgM antibody to MCF-7 tumor cells was observed in seven patients, although there was minimal binding of IgG antibody. Two patients developed significant antibody titers post-high-dose chemotherapy and stem cell reinfusion. There was no evidence of T cell activation. This MUC1-KLH conjugate plus QS-21 was immunogenic and well tolerated in breast cancer patients. Additional trials are ongoing to determine the optimal MUC1 peptide for use in larger clinical trials. Further investigation of vaccine therapy in high-risk breast cancer is warranted. PMID- 10815888 TI - Predicting cancer development in oral leukoplakia: ten years of translational research. AB - Our 10-year translational study of the oral premalignant lesion (OPL) model has advanced the basic understanding of carcinogenesis. Although retinoids have established activity in this model, a substantial percentage of our OPL patients progress to cancer, especially after treatment is stopped. On the basis of our 10 year OPL study, we have developed the first comprehensive tool for assessing cancer risk of OPL patients. This cancer risk assessment tool incorporates medical/demographic variables, epidemiological factors, and cellular and molecular biomarkers. Between 1988 and 1991, 70 advanced OPL patients were enrolled in a chemoprevention trial of induction with high dose isotretinoin (1.5 mg/kg/day for 3 months) followed by 9 months of maintenance treatment with either low dose isotretinoin (0.5 mg/kg/day) or beta-carotene (30 mg/d; total treatment duration, 1 year). We assessed the relationship between cancer risk factors and time to cancer development by means of exploratory data analysis, logrank test, Cox proportional hazard model, and recursive partitioning. With a median follow up of 7 years, 22 of our 70 patients (31.4%) developed cancers in the upper aerodigestive tract following treatment. The overall cancer incidence was 5.7% per year. The most predictive factors of cancer risk are OPL histology, cancer history, and three of the five biomarkers we assessed (chromosomal polysomy, p53 protein expression, and loss of heterozygosity at chromosome 3p or 9p). In the multivariable Cox model, histology (P = 0.0003) and the combined biomarker score of chromosomal polysomy, p53, and loss of heterozygosity (P = 0.0008) are the strongest predictors for cancer development. Retinoic acid receptor beta and micronuclei were not associated with increased cancer risk. We have demonstrated a successful strategy of comprehensive cancer risk assessment in OPL patients. Combining conventional medical/demographic variables and a panel of three biomarkers can identify high risk patients in our sample. This result will need to be validated by future studies. With the identification of high risk individuals, more efficient chemoprevention trials and molecular targeting studies can be designed. PMID- 10815890 TI - Imaging and phase I study of 111In- and 90Y-labeled anti-LewisY monoclonal antibody B3. AB - B3 is a murine monoclonal antibody (mAb) that recognizes a LewisY carbohydrate antigen present on the surface of many carcinomas. An imaging and Phase I trial was performed to study the ability of 111In-mAb B3 to image known metastasis and determine the maximum tolerated dose (MTD), dose-limiting toxicity (DLT), kinetics, and biodistribution of 90Y-mAb B3. Patients (n = 26) with advanced epithelial tumors that express the LewisY antigen were entered. All patients received 5 mCi of 111In-mAb B3 for imaging. 90Y-mAb B3 doses were escalated from 5 to 25 mCi in 5-mCi increments. 111In-mAb B3 and 90Y-mAb B3 were coadministered over a 1-h infusion. Definite tumor imaging was observed in 20 of 26 patients. Sites imaged included lung, liver, bone, and soft tissues. The MTD of 90Y-mAb B3 was determined to be 20 mCi. The DLTs were neutropenia and thrombocytopenia. Tumor doses ranged from 7.7 to 65.1 rad/mCi. 111In- and 90Y-mAb B3 serum pharmacokinetics (n = 23) were found to be similar. The amount of B3 administered (5, 10, and 50 mg) did not alter the pharmacokinetics. Bone marrow biopsies (n = 23) showed 0.0038+/-0.0016% of injected dose/gram for 111In-mAb B3 compared to 0.0046+/-0.0017% of injected dose/gram for 90Y-mAb B3 (P = 0.009). When given to patients with carcinomas that express the LewisY antigen, 111In-mAb B3 demonstrated good tumor localization. The MTD of 90Y-mAb B3 is 20 mCi, with myelosuppression as the DLT. Higher doses of radioactivity need to be delivered to achieve an antitumor effect. Humanized mAb B3 is being developed for evaluation in radioimmunotherapy. A clinical trial to explore the use of higher doses of 90Y-mAb B3 with autologous stem cell support is planned. PMID- 10815889 TI - Radioimmunoguided surgery in colorectal cancer using a genetically engineered anti-CEA single-chain Fv antibody. AB - In radioimmunoguided surgery (RIGS), a radiolabeled antibody is given i.v. before surgery and a hand-held gamma-detecting probe is used to locate tumor in the operative field. The rapid blood clearance and good tumor penetration of single chain Fv antibodies (scFv) offer potential advantages over larger antibody molecules used previously for RIGS. A Phase I clinical trial is reported on RIGS with scFv (MFE-23-his) to carcinoembryonic antigen (CEA). Thirty-four patients undergoing surgery for colorectal carcinoma (17 primary tumors, 16 liver metastases, and 1 anastomotic recurrence) and 1 patient with liver metastases of pancreatic carcinoma received 125I-labeled MFE-23-his scFv (125I-MFE-23-his) 24, 48, 72, or 96 h before operation. 125I-MFE-23-his showed biexponential blood clearance with alpha and beta half-lives of 0.32 and 10.95 h, respectively. The abdomen was scanned during surgery with a hand-held gamma detecting probe (Neoprobe Corp.). 125I-MFE-23-his showed good tumor localization; comparison with histology showed overall accuracy of 84%. Highest median ratios for tumor:normal tissue and tumor:blood were recorded 72 or 96 h after scFv injection for patients undergoing resection of liver metastases. High levels of radioactivity were found in the kidneys. Five patients had grade 1 fever, and three had a grade 1 rise in blood pressure according to the Common Toxicity Criteria. There was a significant correlation between these ratios and those measured in excised tissues using a laboratory gamma counter (P < 0.001). MFE-23-his scFv antibody localizes in CEA producing carcinomas. The short interval between injection and operation, the lack of significant toxicity, and the relatively simple production in bacteria make MFE-23-his scFv suitable for RIGS. PMID- 10815891 TI - Initial clinical trial of a high-affinity retinoic acid receptor ligand (LGD1550). AB - Retinoids mediate their biological response by binding to specific nuclear receptors, including retinoic acid receptors and/or retinoid X receptors. LGD1550 is a high-affinity ligand for all three retinoic acid receptors (alpha, beta, and gamma isoforms) and a potent inhibitor of AP-1, a protein that is closely linked with trophic responses and malignant transformation. We conducted a dose ranging study to evaluate the pharmacokinetics, safety, clinical tolerance, and potential efficacy of this drug in patients with advanced cancer. Twenty-seven patients received oral doses of LGD1550 once per day at doses ranging from 20-400 microg/m2. Skin toxicity was the dose-limiting reaction at the 400 microg/m2 daily dose level. Less prominent reactions included nausea and headache. No major antitumor effects were observed. Pharmacokinetic studies in 15 patients at five dose levels showed that the peak plasma concentration (Cmax) and areas under the plasma concentration-time curve on day 1 were dose-proportional and were similar to values obtained on days 15, 29, and 84. Unlike other retinoids, LGD1550 did not induce its own metabolism, and there was little evidence of drug accumulation. The t1/2 was approximately 5 h after both the initial and repeated doses. We conclude that once-daily doses of LGD1550 of up to 300 microg/m2 are relatively well tolerated. Additional clinical explorations are warranted, especially in patients with cancers of the prostate, thyroid, head and neck, and cervix. PMID- 10815892 TI - Phase I and pharmacological study of weekly administration of the polyamine synthesis inhibitor SAM 486A (CGP 48 664) in patients with solid tumors. European Organization for Research and Treatment of Cancer Early Clinical Studies Group. AB - A single-agent dose-escalating Phase I and pharmacological study of the polyamine synthesis inhibitor SAM 486A was performed. A dosing regimen of four weekly infusions followed by 2 weeks off therapy was studied. Fifty patients were entered into the study. Dose levels studied were 1.25, 2.5, 5, 8, 16, 32, 48, 70, 110, 170, 270, and 325 mg/m2/week. Pharmacokinetic sampling was done on day 1, and trough samples were taken weekly during the first treatment cycle. Pharmacodynamic sampling was done on days 1 and 22. At 325 mg/m2/week, dose limiting toxicity was seen (one patient each with grade 4 febrile neutropenia, grade 3 neurotoxicity, and grade 3 hypotension with syncope and T-wave inversions on electrocardiogram). The recommended dose for further testing was set at 270 mg/m2/week. Infusion time was increased from 10 to 180 min due to facial paresthesias and flushing and somnolence. Drug exposure increased linearly with dose. Mean +/- SD t1,2 at 70-325 mg/m2 doses was 61.4+/-26.2 h, with a large volume of distribution at steady state. In peripheral blood leukocytes, a clear relationship between dose and inhibitory effect on S-adenosylmethionine decarboxylase or changes in intracellular polyamine pools was not recorded. SAM 486A can be administered safely using a dosing regimen of four weekly infusions followed by 2 weeks off therapy. The recommended dose for Phase II studies using this regimen is 270 mg/m2/week. PMID- 10815893 TI - Phase I trial of TALL-104 cells in patients with refractory metastatic breast cancer. AB - The human cytotoxic T-cell line TALL-104 displays antitumor effects in animals with implanted and spontaneous malignancies. A Phase I trial was conducted to determine toxicity of TALL-104 cell therapy in women with metastatic refractory breast cancer. Fifteen patients with metastatic infiltrating ductal (n = 12), lobular (n = 2), or medullary (n = 1) carcinoma received escalating doses of lethally irradiated TALL-104 cells (three patients/group received 10(6), 3 x 10(6), 10(7), 3 x 10(7), and 10(8) cells/kg) for 5 consecutive days (induction course). Patients without progressive disease received monthly maintenance 2-day infusions at the same dose level. Mild grade I/II toxicity developed in 11 patients regardless of cell dose. One grade IV toxicity consequent to hepatic tumor necrosis occurred in a patient given 10(8) cells/kg, 3 weeks after the induction course. Nine patients progressed within 1 month from induction, and five patients had stable disease for 2-6 months. One patient (at 3 x 10(7)/kg) had improvement of liver metastases and ascites, and a second patient (at 10(6)/kg) experienced a dramatic relief in bone pain. Increases in blood natural killer cell activity and levels of IFN-gamma, interleukin-10, and activation markers (soluble interleukin-2 receptor and soluble intercellular adhesion molecule-1) were often seen. Only one patient developed anti-HLA class I antibody responses against TALL-104 cells; specific CTL activity developed in three patients during induction and in four patients during the maintenance boosts. In conclusion, TALL-104 cells were well tolerated by patients with metastatic breast cancer at the doses and regimen tested. The clinical responses observed in this preliminary trial demonstrate that further investigation of TALL-104 cell therapy is warranted. PMID- 10815895 TI - Loss of imprinting and elevated expression of wild-type p73 in human gastric adenocarcinoma. AB - The p73 gene located at 1p36.3 encodes for a protein with significant similarity to p53. To investigate the penetrance of p73 in gastric carcinogenesis, we analyzed the expression, allelotype, and mutation of p73 in five cell lines and 75 tissues. Although extremely low levels of p73 expression were observed in all noncancerous gastric tissues and four of five cell lines, a significant elevation of p73 was detected in 37 of 39 (94.9%) carcinoma tissues. Furthermore, a tumor specific increase of p73 was identified in 14 of 16 (87.5%) matched sets. Allelotyping analysis using a StyI or BanI polymorphism revealed that 5 of 21 (23.8%) informative carcinomas, but none of 19 noncancerous cases, express p73 biallelically, suggesting the transcriptional activation of a silent allele in a subset of cancers. Whereas the transcription of an active allele was markedly induced by serum starvation or clump formation of the cells, treatment with 5-aza 2'deoxycytidine activated a silent allele with a subsequent up-regulation of an active allele, supporting the genomic imprinting and autoregulation of the gene. Allelic deletion or mutation of the gene was not found, and no association of p73 expression with the mutational status of p53 or expression of p21Waf1 was recognized. Taken together, this study argues that p73 is not a target of genetic alteration in gastric carcinogenesis and suggests that overexpression of p73 might be triggered by physiological stresses accompanied with outgrowth of tumors, such as hypoxia or nutrient deprivation. PMID- 10815894 TI - Clinical significance of defective dendritic cell differentiation in cancer. AB - Defective dendritic cell (DC) function has been described previously in cancer patients and tumor-bearing mice. It can be an important factor in the escape of tumors from immune system control. However, the mechanism and clinical significance of this phenomenon remain unclear. Here, 93 patients with breast, head and neck, and lung cancer were investigated. The function of peripheral blood and tumor draining lymph node DCs was equally impaired in cancer patients, consistent with a systemic rather than a local effect of tumor on DCs. The number of DCs was dramatically reduced in the peripheral blood of cancer patients. This decrease was associated with the accumulation of cells lacking markers of mature hematopoietic cells. The presence of these immature cells was closely associated with the stage and duration of the disease. Surgical removal of tumor resulted in partial reversal of the observed effects. The presence of immature cells in the peripheral blood of cancer patients was closely associated with an increased plasma level of vascular endothelial growth factor but not interleukin 6, granulocyte macrophage colony-stimulating factor, macrophage colony-stimulating factor, interleukin 10, or transforming growth factor-beta and was decreased in lung cancer patients receiving therapy with antivascular endothelial growth factor antibodies. These data indicate that defective DC function in cancer patients is the result of decreased numbers of competent DCs and the accumulation of immature cells. This effect may have significant clinical implications. PMID- 10815896 TI - Expression of N-acetylglucosaminyltransferase V in colorectal cancer correlates with metastasis and poor prognosis. AB - N-Acetylglucosaminyltransferase V (GnT-V) is an enzyme that catalyzes beta 1-6 branching of N-acetylglucosamine on asparagine-linked oligosaccharides of cell proteins. Metastatic potential of various cancer cells has been shown to correlate with increase of GnT-V activity and concomitant beta 1-6 branching of N acetylglucosamine. However, protein expression of GnT-V in human cancer tissue and its clinical significance have not yet been demonstrated. To clarify the possible relationship between metastasis and GnT-V in human colorectal cancer, protein expression of GnT-V was studied using surgically resected specimens. We established a monoclonal antibody against GnT-V and performed immunohistochemical analysis of 103 human colorectal cancer cases. Of 103 cases, 26 cases (25.2 %) showed specific expression of GnT-V in colorectal cancer tissues. The expression of GnT-V was significantly correlated with distant metastasis (P < 0.05, chi2 test). Overall 5-year survival rate was 52.8% for GnT-V-positive patients and 81.7% for GnT-V-negative patients (P < 0.01, Log-rank test). We showed direct evidence for the relationship between GnT-V and metastasis in human colorectal cancer. Screening of GnT-V expression in colorectal cancer may provide useful information for prognosis of postoperative patients. PMID- 10815897 TI - Expression of a novel factor, com1, in early tumor progression of breast cancer. AB - Tumor cells and their surrounding microenvironment produce a variety of growth factors and proteolytic enzymes to promote tumor growth and metastasis. We have recently identified a novel factor, termed com1, which is up-regulated in human breast carcinoma cells upon formation of experimental metastatic tumors and assumed to act as a growth-promoting factor in breast cancer. In attempts to explore the biological role of com1 in clinical tumor growth and metastasis, expression of com1 mRNA in primary carcinomas from 81 breast cancer patients and 27 samples of uninvolved adjacent breast tissue from these patients was compared and related to known prognostic parameters and outcome. The levels of com1 mRNA were significantly up-regulated (P < 0.0001) in the tumors compared to the normal breast tissues. Tumor expression of com1 mRNA, however, did not correlate with the mRNA levels for urokinase-type plasminogen activator (uPA), its receptor, or the type 1 inhibitor, which are factors that define a phenotype of tumor aggressiveness when elevated. And whereas the mRNA levels of uPA and the uPA receptor were elevated in tumors from the patients who subsequently had poor outcome, no correlations were observed between tumor com1 mRNA expression and prognosis or histological and biochemical characteristics of the tumors. We therefore assume that com1 may mediate some growth-promoting function early in development of the primary breast carcinoma, but not in later stages of tumorigenesis or metastasis. PMID- 10815898 TI - Microsatellite instability and expression of hMLH-1 and hMSH-2 in sebaceous gland carcinomas as markers for Muir-Torre syndrome. AB - Sebaceous gland carcinomas (SGCs) are rare malignant skin tumors occurring sporadically or as a phenotypic feature of the Muir-Torre syndrome (MTS). A subset of patients with MTS have a variant of the hereditary nonpolyposis colorectal cancer syndrome caused by mutations in mismatch repair (MMR) genes, which lead to microsatellite instability (MSI). We evaluated the value of MSI and loss of expression of the MMR genes, hMLH-1 and hMSH-2, as a marker to identify and distinguish MTS from sporadic SGC. Using a nationwide pathology report database system, we identified patients with the MTS phenotype. SGCs from 10 MTS patients and the colorectal carcinomas from 3 additional MTS patients were collected. In addition, SGCs from eight patients without a history of visceral neoplasm were collected. MSI was detected in 9 of 13 MTS-associated tumors (69%) versus 0 of 8 sporadic SGCs (P = 0.002). Except for the age of onset of colorectal carcinoma [58 years in the MSI-positive group versus 69.8 years in the MSI-negative group (P = 0.17)], no differences were seen between the MSI-negative and the MSI-positive MTS patients. Loss of expression of hMLH-1 (n = 4) or hMSH-2 (n = 4) was found in MSI-positive patients only. MSI and loss of expression of MMR genes can be used as markers for MTS in patients with SGC. Consequently, MSI and loss of MMR gene expression in a patient presenting with SGC as the initial malignancy have important consequences for the patient and family. There are at least two variants of MTS with different molecular genetic mechanisms because 31% of the patients with the MTS phenotype had no MSI. PMID- 10815899 TI - Prognostic value of serial tissue prostate-specific antigen measurements during different hormonal treatments in prostate cancer patients. AB - To reveal the effects of different hormonal treatments directly on the prostate during treatment, the concentration of prostate-specific antigen in the tissue (T PSA) was studied in 63 patients with untreated newly diagnosed carcinoma of the prostate (CaP). T-PSA measurements were performed in fine-needle aspiration biopsies at the time of diagnosis and 6, 12, and 24 months after initiation of treatment. Treatments modalities were bilateral orchidectomy, gonadotropin releasing hormone (GnRH) agonists, or parenteral estrogens. Thirty-one (49%) of the patients died of CaP and 18 (29%) of other diseases. Fourteen of the patients (22%) were still alive at the end of the observation period (median follow-up time, 111.5 months; range, 98-128 months). In all of the 31 patients who died of CaP, T-PSA values increased during treatment. This increase was observed long before clinical signs of progression appeared (median of interval, 14 months). Twenty of these 31 patients showed an increase in T-PSA from pretreatment values at 6 months. At 12 months this increase was observed in 30 of 31 patients. In contrast, in all of the patients who responded to the hormonal regimen, T-PSA values decreased and remained low during treatment. Furthermore, the patients who did not die of CaP and received estrogen treatment had significantly higher T-PSA values compared with those who were treated with bilateral orchidectomy or GnRH agonists. This indicates that estrogens may stimulate PSA synthesis in tumor tissue in vivo in the presence of castration levels of testosterone. Statistical evaluation showed that the T-PSA ratio between month 12 and month 0 had the most significant prognostic value for predicting the clinical outcome. This ratio was superior to clinical classifications, e.g., tumor stage and cytological grade, and also was higher than T-PSA at the time of diagnosis. This study has shown that aspiration biopsy material can be used to reveal biochemical changes in the tissue during treatment and that one specific marker (T-PSA) can predict the clinical outcome of endocrine treatment of CaP patients better than previously used methods. We believe that selected tissue markers or the protein pattern can help us to characterize the tumors and predict the clinical outcome so an optimal treatment can be chosen for every patient. PMID- 10815900 TI - Expression and prognostic significance of IAP-family genes in human cancers and myeloid leukemias. AB - Expression of several inhibitor of apoptosis proteins (IAPs) was investigated in the National Cancer Institute panel of 60 human tumor cell lines, and the expression and prognostic significance of one of these, XIAP, was evaluated in 78 previously untreated patients with acute myelogenous leukemia (AML). XIAP and cIAP1 were expressed in most cancer lines analyzed, with substantial variability in their relative levels. In contrast, NAIP mRNA was not detectable, and cIAP2 was found at the mRNA and protein levels in only 34 (56%) and 5 (8%) of the 60 tumor cell lines analyzed, respectively. Interestingly, XIAP, cIAP1, and cIAP2 mRNA levels did not correlate with protein levels in the tumor lines, indicating posttranscriptional regulation of expression. High levels of XIAP protein in tumor cell lines were unexpectedly correlated with sensitivity to some anticancer drugs, particularly cytarabine and other nucleosides, whereas higher levels of cIAP1 protein levels were associated with resistance to several anticancer drugs. The relevance of XIAP to in vivo responses to cytarabine was explored in AML, making correlations with patient outcome (n = 78). Patients with lower levels of XIAP protein had significantly longer survival (median, 133 versus 52.5 weeks; P = 0.05) and a tendency toward longer remission duration (median, 87 versus 52.5 weeks; P = 0.13) than those with higher levels of XIAP. Altogether, these findings show that IAPs are widely but differentially expressed in human cancers and leukemias and suggest that higher XIAP protein levels may have adverse prognostic significance for patients with AML. PMID- 10815901 TI - Ras activation in normal white blood cells and childhood acute lymphoblastic leukemia. AB - Ras is an important cellular switch, relaying growth-promoting signals from the plasma membrane to the nucleus. In cultured cells, Ras is activated by various hematopoietic cytokines and growth factors, but the activation state of Ras in peripheral WBCs and bone marrow cells has not been studied nor has Ras activation been assessed in cells from patients with acute lymphoblastic leukemia (ALL). Using an enzyme-based method, we assessed Ras activation in peripheral WBCs, lymphocytes, and bone marrow cells from normal subjects and from children with T cell ALL (T-ALL) and B-lineage ALL (B-ALL). In normal subjects, we found mean Ras activations of 14.3, 12.5, and 17.2% for peripheral blood WBCs, lymphocytes, and bone marrow cells, respectively. All three of these values are higher than we have found in other normal human cells, compatible with constitutive activation of Ras by cytokines and growth factors present in serum and bone marrow. In 9 of 18 children with T-ALL, Ras activation exceeded two SDs above the mean of the corresponding cells from normal subjects, whereas in none of 11 patients with B ALL did Ras show increased activation; activating genetic mutations in ras occur in less than 10% of ALL patients. Thus, Ras is relatively activated in peripheral blood WBCs, lymphocytes, and bone marrow cells compared with other normal human cells, and Ras is activated frequently in T-ALL but not in B-ALL. Increased Ras activation in T-ALL compared with B-ALL may contribute to the more aggressive nature of the former disease. PMID- 10815902 TI - Telomerase activity and expression of hTRT and hTR in gastrointestinal stromal tumors in comparison with extragastrointestinal sarcomas. AB - Stromal tumors of the gut (GISTs) have rarely been analyzed for genetic alterations. This study aimed at determining telomerase activity and the expression of the telomerase subunits human telomerase reverse transcriptase (hTRT) and human telomerase RNA (hTR) in GISTs and extragastrointestinal neurogenic or myogenic sarcomas. Telomerase activity was investigated using the telomeric repeat amplification protocol assay in 21 GISTs, recurrences and liver metastases from 16 patients, and in 22 leiomyosarcomas and 21 malignant peripheral nerve sheath tumors (MPNSTs), which served as reference tumors. Expression of hTRT and hTR mRNA was investigated using reverse transcription-PCR. Thirteen GISTs were localized in the stomach and three in the small intestine. Two tumors were benign. In one case, the biological behavior was uncertain. In 67% of GISTs, high telomerase activity was found, whereas high activity was noted in only 18% of leiomyosarcomas and in 48% of MPNSTs. There was no activity in two benign and two malignant GISTs. In one malignant tumor of the small intestine, the primary tumor showed no activity at first but a marked activity in its recurrence. In the tumor with uncertain behavior, telomerase activity and hTRT expression were only weak. In all GISTs showing telomerase activity, the catalytic subunit hTRT was expressed. All GISTs and extragastrointestinal sarcomas expressed hTR. In comparison with leiomyosarcomas and MPNSTs, malignant GISTs showed a higher telomerase activity, which, however, was not seen in benign GISTs. It is possible that telomerase activity occurs during the progression of malignant GISTs. There was a correlation between telomerase activity and the expression of hTRT. PMID- 10815903 TI - Association of allelic loss on 1q, 4p, 7q, 9p, 9q, and 16q with postoperative death in papillary thyroid carcinoma. AB - Papillary thyroid carcinomas, most of which are characterized by slow growth and good prognosis, account for the majority of thyroid carcinomas. To provide appropriate postoperative management, it is important to classify them by prediction of their prognosis. To find genetic markers associated with poor prognosis, allelic loss at all 39 nonacrocentric chromosome arms was compared in 24 deceased cases and 45 age-, sex-, stage-, and type-matched survived cases. Allelic loss was examined in primary tumors from both groups using highly polymorphic microsatellite markers on 39 nonacrocentric autosomal arms. Age at diagnosis, sex, stage, and types of tumors were matched between the two groups. No recurrent tumor was used for DNA analysis. Mean fractional allelic loss in the deceased and survived cases was 0.10+/-0.08 and 0.03+/-0.05 (P < 0.001). The survived cases showed marginal frequencies of allelic loss throughout all chromosome arms except 22q. The deceased cases showed frequent allelic losses on chromosomes 1q (37%), 4p (21%), 7q (20%), 9p (36%), 9q (31%), and 16q (29%), with significant difference (P < 0.05). These chromosome regions may include tumor suppressor genes whose inactivation is associated with aggressive phenotypes of papillary thyroid carcinoma. PMID- 10815904 TI - Microregional heterogeneity of non-protein thiols in cervical carcinomas assessed by combined use of HPLC and fluorescence image analysis. AB - Under low oxygen conditions, non-protein thiols (NPSHs, non-protein sulfhydryls) can effectively compete for DNA radicals sites and hence represent a potentially important cause of radiation resistance in the clinic. Intra- and intertumoral heterogeneity of glutathione (GSH) and cysteine were assessed in cryostat sections of multiple biopsies obtained from 10 cervical carcinomas by the combined use of a sensitive high-performance liquid chromatography (HPLC) method and a fluorescence image analysis technique to examine the spatial distribution of NPSHs in tumor tissue. Glutathione concentrations ranged from 1.98 to 4.42 mM; significant (> or =1 mM) concentrations of cysteine, a more effective radioprotector than GSH, were found in some tumors. By HPLC, the intratumoral heterogeneity of NPSHs was relatively small compared with the intertumoral heterogeneity. The histochemical stain 1-(4-chloromercuryphenoylazo)-2-napthol (mercury orange), which binds to GSH and cysteine, was used to determine the spatial distribution of NPSHs in tumor tissue. A comparison of NPSH levels in serial cryostat sections showed a close correlation between NPSH values determined by HPLC and mercury orange fluorescence quantification. Using fluorescence image analysis, an approximately 2-fold increase of NPSHs in tumor versus nonmalignant tissue was observed in the same section. Because some cervical carcinomas contain radiobiologically important levels of cysteine, agents that target the biochemical pathways maintaining tumor cysteine have therapeutic potential as adjuncts to radiotherapy in cervix cancer patients. PMID- 10815905 TI - Frequent amplification of chromosomal region 20q12-q13 in ovarian cancer. AB - DNA amplification at chromosomal region 20q12-q13, which is common in breast cancer, has recently been described also in ovarian tumors. We studied the amplification of the recently identified candidate oncogenes in this region in 24 sporadic, 3 familial and 4 hereditary ovarian carcinomas, and in 8 ovarian cancer cell lines. High-level amplification of at least one of the five nonsyntenic regions at 20q12-q13.2 was found in 13 sporadic (54%) and in all four hereditary tumors. Typically, two or more distinct amplicons (separated by nonamplified DNA) were found coamplified in various combinations. The regions defined by the AIB1 and PTPN1 genes (at 20q12 and 20q13.1, respectively) were amplified in 25% and 29% of the sporadic tumors, also without simultaneous coamplification of other regions. Amplification of AIB1 (a steroid receptor coactivator gene) was associated with estrogen receptor positivity in sporadic ovarian carcinomas (P = 0.01) and showed a tendency to correlate with poor survival of patients. Of the genes amplified in breast cancer, the BTAK gene was amplified in 21%, the MYBL2 gene in 17%, and the ZNF217 gene in 12.5% of the sporadic tumors. The high frequency of gene amplification at 20q12-q13.2 suggests that the genes amplified therein may play a central role in the pathogenesis of sporadic and hereditary ovarian carcinoma. PMID- 10815906 TI - mdm2 expression as a prognostic indicator in clear cell renal cell carcinoma: comparison with p53 overexpression and clinicopathological parameters. AB - The present study was designed to analyze the expression of p53 and mdm2 in clear cell renal cell carcinoma with special emphasis on their association with tumor grade and clinical outcome. In particular, the value of individual protein overexpression as well as combined p53/mdm2 positivity was evaluated because both proteins are functionally connected, and their expression is controlled by an autoregulatory feedback loop. A cohort of 97 clear cell renal cell carcinomas was analyzed. The overexpression of mdm2 and p53 proteins was investigated on paraffin-embedded material by using monoclonal antibodies. Eighteen tumors showed mdm2 positivity, whereas 35 of the tumors overexpressed p53. Whereas p53 and mdm2 positivity correlated significantly (P = 0.00004), no correlation could be found between mdm2 protein overexpression and tumor stage, lymph node involvement, and presence of distant metastases. mdm2 positivity was found significantly more frequently in tumors of higher grade. In univariate analysis, there was a statistically significant correlation between p53 and mdm2 overexpression in the same tumor and poor survival (P = 0.00179). Multivariate analysis revealed that coincident mdm2/p53 overexpression, the presence of distant metastases, and tumor grade were independent predictors for tumor progression. Our results indicate that mdm2/p53 co-overexpression, nuclear grade, and preoperative presence of distant metastasis are independent predictors for poor survival. PMID- 10815907 TI - Loss of nuclear p16 protein expression correlates with increased tumor cell proliferation (Ki-67) and poor prognosis in patients with vertical growth phase melanoma. AB - The CDKN2A (p16INK4alpha) cell cycle-inhibitory gene has been associated with development of familial melanoma. Additionally, recent studies indicate that p16 alterations occur frequently in sporadic melanomas. To investigate whether differences in p16 expression are associated with tumor cell proliferation, tumor progression, and patient survival, we examined the immunohistochemical staining of p16 protein in a consecutive series of 202 vertical growth phase melanomas and 68 corresponding metastases and compared the results with Ki-67 expression, p53 expression, clinicopathological variables, and survival data. Forty-five percent of the primary tumors showed absent or minimal nuclear staining for p16 protein. These cases were significantly associated with high Ki-67 expression (P < 0.0001), ulceration (P = 0.001), and vascular invasion (P = 0.03). Further loss of p16 expression was observed in metastatic lesions (77% were negative; P < 0.0001). Absent/minimal nuclear p16 staining significantly predicted poor patient survival (log-rank test, P = 0.0003), with 37% and 67% estimated 10-year survival rates for cases with absent or present p16 expression, respectively. In multivariate analysis, p16 staining was an independent prognostic factor (hazard ratio, 2.5; 95% confidence interval, 1.5-4.2; P = 0.0008), along with p53 expression, Ki-67 expression, anatomical site, Clark's level of invasion, and vascular invasion. Our findings indicate that loss of nuclear p16 protein expression in vertical growth phase melanomas is associated with increased tumor cell proliferation (Ki-67) and independently predicts decreased patient survival. Cases without p53 expression had improved survival. PMID- 10815908 TI - Reproducibility of p53 immunohistochemistry in bladder tumors. National Cancer Institute, Bladder Tumor Marker Network. AB - The National Cancer Institute Bladder Tumor Marker Network conducted a study to evaluate the reproducibility of immunohistochemistry for measuring p53 expression in bladder tumors. Fifty paraffin blocks (10 from each of the five network institutions) were chosen at random from among high-grade invasive primary bladder tumors. Two sections from each block were sent to each laboratory for staining and scoring, and then all sections were randomly redistributed among the laboratories for a second scoring. Intra- and interlaboratory reproducibility was assessed with regard to both staining and scoring. For overall assessments of p53 positivity, the results demonstrated that intralaboratory reproducibility was quite good. Concordance across the five participating laboratories was high for specimens exhibiting no or minimal nuclear immunostaining of tumor cells or high percentages of tumor cells with nuclear immunoreactivities. However, there was a reduced level of concordance on specimens with percentages of stained tumor cells in an intermediate range. The discordancies were due mainly to staining differences in one of the five laboratories and scoring differences in another laboratory. These results indicate that some caution must be used in comparing results across studies from different groups. Standardization of staining protocols and selection of a uniform threshold for binary interpretation of results may improve assay reproducibility between laboratories. PMID- 10815909 TI - Immunohistochemical determination of five somatostatin receptors in meningioma reveals frequent overexpression of somatostatin receptor subtype sst2A. AB - Meningioma is one of a variety of human tumors that exhibit a very high density of somatostatin receptors and in many cases show a true positive somatostatin receptor scintigraphy. However, the level of expression of individual somatostatin receptor proteins in meningioma has not been investigated. We have recently developed a panel of somatostatin receptor subtype-specific antibodies that effectively stain formalin-fixed, paraffin-embedded tumor tissue (S. Schulz et al., Clin. Cancer Res., 4: 2047-2052, 1998). In the present study, we have used these antibodies to determine the somatostatin receptor status of 40 randomly selected meningiomas. Immunoreactive staining for all somatostatin receptors was clearly located at the plasma membrane of the tumor cells and completely blocked with antigenic peptide. The vast majority of tumors (29 cases; 70%) were positive for sst2A immunoreactivity; among these, 20 (69%) tumors showed high levels of sst2A immunoreactivity. In contrast, all other somatostatin receptors were only detected sporadically, and none of these cases revealed a particularly strong staining. However, it is uncertain to what extent somatostatin receptor-immunoreactive staining intensity may translate into somatostatin receptor protein expression on the tumor cells. Therefore, in a prospective study, 16 surgically removed meningiomas were collected, and the level of sst2A expression was determined using Western blot analysis. Whereas sst2A was readily detectable as a broad band migrating at Mr 70,000 in 12 (75%) of these tumors, 8 tumors (50%) showed particularly high levels of immunoreactive sst2A receptors. There was an excellent correlation (P < 0.001) between the level of sst2A protein expression detected in Western blots and the sst2A- immunoreactive staining seen in tissue sections. Thus, the frequent overexpression of the sst2A receptor may explain the high tracer uptake often observed in meningioma patients during somatostatin receptor scintigraphy. Moreover, this simple immunohistochemical method could prove useful in identifying those cases of recurrent disease that may possibly respond to therapy with sst2-selective agonists. PMID- 10815910 TI - A high number of tumor-infiltrating lymphocytes are associated with a small tumor size, low tumor stage, and a favorable prognosis in operated small cell lung carcinoma. AB - Tumor-infiltrating lymphocytes, apoptosis, and angiogenesis have a pivotal role in tumor growth control. This study was undertaken to analyze the associations of these factors and their role in the prognosis, defined as survival time, of 56 patients operated on for small cell lung carcinoma (SCLC). Immunohistochemically detected T cells and macrophages were the most abundant tumor-infiltrating lymphocytes in SCLC, whereas the number of B cells was small. There was a trend in the number of intratumoral cytotoxic/suppressor CD8 cells that were associated with the extent of apoptotic bodies in SCLC, as measured by in situ 3'-end labeling of apoptotic DNA. A high number of intratumoral T cells and CD8 cells were associated significantly with a low tumor size (<3 cm) and low tumor stage (stages I-II). A high number of intratumoral macrophages were associated with a low tumor stage and angiogenesis, as measured by microvessel density. A high number of T cells, CD8 cells, and macrophages and a low tumor size (<3 cm) were prognostic markers predicting favorable survival time of the patients with SCLC. PMID- 10815911 TI - Association between immunohistochemical expression of vascular endothelial growth factor (VEGF), VEGF-expressing neuroendocrine-differentiated tumor cells, and outcome in prostate cancer patients subjected to watchful waiting. AB - Tumor growth is dependent on angiogenesis, which is thought to be controlled by angiogenic factors. Therefore, the immunoreactivity of the angiogenic cytokine vascular endothelial growth factor (VEGF) was semiquantitatively scored in archival prostate tumors obtained at diagnosis in 221 patients followed expectantly. At diagnosis, 125 patients suffered from clinically localized disease. Median length of follow-up was 15 years, and 57% of the patients eventually died of prostate cancer. All of the tumors exhibited cytoplasmic staining for VEGF. The staining intensity was weak in 47 tumors and moderate and strong in 107 and 67, respectively. VEGF expression was significantly correlated with microvessel density (MVD; median, 43; range, 16-151; P = 0.014), increasing T-classification (P = 0.001), dedifferentiation (P < 0.001), and disease-specific survival (P = 0.013). Strongly VEGF-immunoreactive, neuroendocrine-differentiated (NE) tumor cells were observed in 125 tumors. NE expression was significantly correlated with increasing MVD, increasing T-classification, dedifferentiation, and survival (all, P < 0.001). MVD and NE tumor cell expressions were significant variables in a multivariate analysis that included patients with clinically localized prostate cancer only. VEGF and NE expression were significantly correlated with MVD, clinical characteristics, and disease-specific survival. NE expression was a significant prognostic marker in localized prostate cancer patients, whereas the applied semiquantitatively scoring of VEGF expression was inadequate to make this growth factor provide any additional prognostic information. Moreover, the significant VEGF expression of NE tumor cells suggests an additional important character of these cells in the involvement in disease progression. PMID- 10815912 TI - Overexpression of cyclin D1 is associated with metastatic prostate cancer to bone. AB - Cyclin D1 is a key regulator of the G1 phase progression of the cell cycle. There is increasing evidence that deregulated cyclin D1 expression is implicated in tumorigenesis and tumor progression in certain neoplasms. Recently, it has been reported that cyclin D1 overexpression might be related to the evolution of androgen-independent disease in prostate cancer. This study was conducted to investigate patterns of cyclin D1 expression in prostate cancer samples representing different points in the natural history and treatment evolution of the disease. Association with clinical outcomes was also explored. Using immunohistochemistry, 86 radical prostatectomy specimens (53 naive and 33 after androgen deprivation) and 22 androgen-independent bone metastases were studied. We examined the difference in cyclin D1 expression in primary versus metastatic cases. In addition, we examined the association in primary cases between cyclin D1 expression and clinicopathological parameters of poor clinical outcome, including time to prostate-specific antigen relapse and Ki67 proliferative index. Cyclin D1-positive phenotype, defined as identification of positive immunoreactivity in the nuclei of > or =20% of tumor cells, was observed in 10 of 86 (11%) primary cases compared with 15 of 22 (68%) androgen-independent bone metastases (P = 0.001). There was no correlation between cyclin D1 overexpression and either Gleason score, neo-adjuvant hormone treatment, or prostate-specific antigen relapse We observed a statistical association between cyclin D1 overexpression and high Ki67 proliferative index, defined as > or =20% of positive tumor cells (P = 0.02). These data support the hypothesis that cyclin D1 overexpression may represent an oncogenic event in androgen-independent metastatic prostate cancer to the bone. PMID- 10815913 TI - The cell cycle inhibitors p21WAF1 and p27KIP1 are associated with survival in patients treated by salvage prostatectomy after radiation therapy. AB - We evaluated p27KIP1 and p21WAF1 expression in 52 patients treated by salvage radical prostatectomy and bilateral pelvic lymphadenectomy for biopsy-proven locally persistent or recurrent prostate cancer after external beam radiation therapy. We defined low and high expression based on the median value observed in our sample. Five-year distant metastasis-free survival and cancer-specific survival were 71 and 82%, respectively, for patients with low expression of p21 (< or =5%), compared with 94 and 100%, respectively, for those with high expression of p21 (>5%; P = 0.02 and 0.01, respectively). Five-year distant metastasis-free survival and cancer-specific survival were 71 and 82%, respectively, for patients with low expression of p27 (<50%), compared with 88 and 96%, respectively, for those with high expression of p27 (> or =50%; P = 0.06 and 0.01, respectively). These findings indicate that p21 and p27 expression levels are significant predictors of survival for patients selected for salvage prostatectomy for recurrent prostate cancer. PMID- 10815914 TI - High-level expression of angiogenic factors is associated with advanced tumor stage in human neuroblastomas. AB - Angiogenesis is essential for tumor growth and metastasis and depends on the production of angiogenic factors by tumor cells. Neuroblastoma (NB) is a common pediatric tumor of neural crest origin, which is biologically and clinically heterogeneous. Increased tumor vascular index correlates with poor outcome of NB. To determine which angiogenic factors contribute to NB angiogenesis and thereby support tumor progression, we examined the expression of eight angiogenic factors [vascular endothelial growth factor (VEGF), VEGF-B, VEGF-C, basic fibroblast growth factor, angiopoietin (Ang)-1, Ang-2, transforming growth factor alpha, and platelet-derived growth factor (PDGF)] by semiquantitative RT-PCR in 37 NB primary tumors and in 22 NB cell lines. We also analyzed the relationship between angiogenic factor expression and clinicopathological factors as well as patient survival. All eight angiogenic factors examined were expressed at various levels in NB cell lines and tumors, suggesting their involvement in NB angiogenesis. The expression levels of most angiogenic factors were correlated with each other, suggesting their synergy in regulating the angiogenic process. Significantly higher expression levels of VEGF, VEGF-B, VEGF-C, basic fibroblast growth factor, Ang-2, transforming growth factor alpha, and PDGF-A (P < 0.0001-0.026) were found in advanced-stage tumors (stages 3 and 4) compared with low-stage tumors (stages 1, 2, and 4S). Expression of PDGF-A was significantly associated with patient survival (P = 0.04). The redundancy in angiogenic factor expression suggests that inhibition of VEGF bioactivity alone might not be a sufficient approach for antiangiogenic therapy of human NB. PMID- 10815915 TI - Mucin core protein expression in colorectal cancers with high levels of microsatellite instability indicates a novel pathway of morphogenesis. AB - Particular mucinous phenotypes have been associated with serrated epithelial polyps of the colon. These polyps also show a high frequency of DNA instability. The aim of this study was to examine the expression of mucins in colorectal cancers that arise through the suppressor and mutator pathways. The immunohistochemical distribution of the human apomucins MUC1, MUC2, MUC4, and MUC5AC was determined in 93 sporadic colorectal cancers classified previously (J. R. Jass et al., J. Clin. Pathol., 52: 455-460, 1999) according to levels of DNA microsatellite instability (MSI) as 22 MSI-high (MSI-H), 24 MSI-low (MSI-L), and 47 MS stable (MSS). MUC2 expression was observed in 19 (86%) MSI-H, 10 (42%) MSI L, and 15 (32%) MSS cancers (P = 0.0001); and MUC5AC expression was observed in 17 (77%) MSI-H, 8 (33%) MSI-L, and 13 (28%) MSS cancers (P = 0.0003). There was no association between MUC1 or MUC4 expression and MSI status. The mucinous phenotype described in serrated polyps (MUC2+/MUC5AC+) was seen in 15 (68%) of 22 MSI-H and only 10 (14%) of 71 MSI-L/MSS cancers (P < 0.0001). Increased expression of the secretory mucins MUC2 and MUC5AC was observed in sporadic MSI-H cancers. Identical mucin changes and DNA MSI occurred in serrated polyps of the colorectum, which suggests that these lesions may represent precursors of MSI-H cancers. PMID- 10815916 TI - Coexpression of MUC1 glycoprotein with multiple angiogenic factors in non-small cell lung cancer suggests coactivation of angiogenic and migration pathways. AB - We investigated the expression of MUC1 protein and its relationship to the microvessel density and the expression of thymidine phosphorylase, vascular endothelial growth factor (VEGF), VEGF-receptor KDR, basic fibroblast growth factor (bFGF), and bFGF-receptor (FGFR-2) in non-small cell lung cancer. Although MUC1 expression was found equally in poorly and highly vascularized tumors, a significant coexpression with multiple angiogenic factors and their receptors was noted (P = 0.0002, 0.03, 0.19, 0.10, and 0.01 for thymidine phosphorylase, VEGF, KDR, bFGF, and FGFR-2, respectively). In multiple regression analysis, both angiogenesis and MUC1 expression were independent prognostic variables. The present study suggests the existence of an early genetic event leading to the activation of both migration and angiogenesis pathways in non-small cell lung cancer. PMID- 10815917 TI - Flow cytometric analysis of androgen receptor expression in human prostate tumors and benign tissues. AB - Androgen receptor (AR) plays an important role in growth and hormonal therapy of human prostate tumors. Immunohistochemical analysis of AR expression, a nonquantitative technique, is currently used for screening of receptor expression in prostate tissues. The present report describes a laser flow cytometric method for monitoring AR expression in human cell lines and in archival formalin-fixed paraffin-embedded prostate tissues and tumors. Multiparametric flow analysis can be used for simultaneous detection of other cellular markers (e.g., DNA aneuploidy), and by gated analysis, AR expression in subpopulations of a tumor can be quantitatively determined. PMID- 10815918 TI - Differential recognition of a BCR/ABL peptide by lymphocytes from normal donors and chronic myeloid leukemia patients. AB - The BCR/ABL oncogenic fusion protein transforms normal bone marrow stem cells into neoplastic cells. It has been shown that peptides derived from the junctional region of this oncogenic fusion protein can be recognized by human T lymphocytes obtained from normal donors. In this study, we investigated the immunogenicity in patients with chronic myeloid leukemia (CML) of a 17 mer b3/a2 Bcr/abl peptide (B/A1), which was shown to induce proliferative responses in lymphocytes from normal donors. A total of 56 CML patients in chronic phase were studied. Twenty-two patients were studied at diagnosis without any treatment (group I). Fourteen patients were receiving IFN (group II), 14 patients were being treated with hydroxyurea (group III), and 6 patients were on different regimens (group IV). Patients were initially assessed for general immunological competence using both in vivo and in vitro assays. Patients were also selected for the expression of HLA-DR0401, the HLA specificity known to present peptide B/A1 to CD4 lymphocytes. With the exception of the six patients in group IV, the results of all these assays (in vitro phytohemagglutinin/tetanus toxoid responses, in vivo skin reaction to ubiquitous antigens) in CML patients did not significantly differ from those obtained in normal donors, thus excluding the presence of generalized immunosuppression. Eight patients with HLA-DR0401 and a b3/a2 type of fusion were identified and further studied. In these eight patients dendritic cells were obtained from adherent peripheral blood mononuclear cells and used to stimulate CD4 lymphocytes. No patient developed a specific response to the bcr/abl peptide, although patients' lymphocytes proliferated in response to a promiscuous tetanus toxoid peptide in all but one case. In contrast, response to the bcr/abl peptide was observed in seven of eight HLA-DR0401 healthy donors tested. These data suggest that immunocompetent, HLA-DR0401+ CML patients are unable to respond to peptide B/A1, at difference from healthy donors. The implication of these results for the immunotherapy of CML is discussed. PMID- 10815919 TI - Epidermal growth factor receptor blockade with C225 plus gemcitabine results in regression of human pancreatic carcinoma growing orthotopically in nude mice by antiangiogenic mechanisms. AB - Both epidermal growth factor receptor (EGF-R) signaling mechanisms and angiogenesis have been evaluated as independent targets for therapy of human pancreatic carcinoma, but a link between the two processes has been identified only recently. This study evaluated whether EGF-R blockade therapy with anti-EGF R antibody C225 inhibits pancreatic carcinoma growth and metastasis in an orthotopic nude mouse model via tumor-mediated angiogenesis and whether gemcitabine potentiates this effect. In vitro treatment of human pancreatic carcinoma L3.6pl cells with C225 inhibited EGF-R autophosphorylation, producing a maximum of 20% cytostasis. Treatment with C225 plus gemcitabine resulted in additive cytotoxic effects that increased with increasing gemcitabine concentrations. Dose-dependent decreases in expression of the angiogenic factors vascular endothelial growth factor and interleukin 8 (but not basic fibroblast growth factor) were observed in the C225-treated cells (mRNA and protein levels). In L3.6pl tumors established in the pancreas of nude mice, systemic therapy with C225 alone and C225 in combination with gemcitabine resulted in growth inhibition, tumor regression, and abrogation of metastasis; median tumor volume was reduced from 538 to 0.3 and to 0 mm3, respectively. Gemcitabine treatment alone reduced median tumor volume from 538 to 152 mm3. Liver metastases were present in 50% of the controls, 30% of the gemcitabine-treated animals, and 20% of C225-treated animals. No macroscopically visible liver metastases were observed in the combination treatment group. As early as 11 days after C225 treatment, the median percentage of proliferating cell nuclear antigen-positive cells was substantially reduced compared with gemcitabine treatment alone (26% versus 73%, respectively) versus controls (92%), correlating with in vivo blockade of EGF-R activation. Similarly after 11 days treatment, production of vascular endothelial growth factor and interleukin 8 was significantly lower in C225 and C225 plus gemcitabine-treated tumors versus gemcitabine-treated and control tumors. Significant differences in microvessel density were observed 18 days after C225 or combination treatments (but not gemcitabine alone) in direct correlation with the difference in percentage of apoptotic endothelial cells, as visualized by double immunofluorescence microscopy. These experiments indicate that therapeutic strategies targeting EGF-R have a significant antitumor effect on human L3.6pl pancreatic carcinoma growing in nude mice which is mediated in part by inhibition of tumor-induced angiogenesis, leading to tumor cell apoptosis and regression. Furthermore, this effect is potentiated in combination with gemcitabine. PMID- 10815920 TI - Nuclear delivery of doxorubicin via folate-targeted liposomes with bypass of multidrug-resistance efflux pump. AB - Folic acid, attached to polyethyleneglycol-derivatized, distearoyl phosphatidylethanolamine, was used to target in vitro liposomes to folate receptor (FR)-overexpressing tumor cells. Confocal fluorescence microscopic observations demonstrated binding and subsequent internalization of rhodamine labeled liposomes by a high FR-expressing, murine lung carcinoma line (M109-HiFR cells), with inhibition by free folic acid. Additional experiments tracking doxorubicin (DOX) fluorescence with DOX-loaded, folate-targeted liposomes (FTLs) indicate that liposomal DOX is rapidly internalized, released in the cytoplasmic compartment, and, shortly thereafter, detected in the nucleus, the entire process lasting 1-2 h. FR-mediated cell uptake of targeted liposomal DOX into a multidrug resistant subline of M109-HiFR cells (M109R-HiFR) was unaffected by P glycoprotein-mediated drug efflux, in sharp contrast to uptake of free DOX, based on verapamil-blockade experiments with quantitation of cell-associated DOX and flow cytometry analysis. Delivery of DOX by FTLs to M109R-HiFR cells increased continuously with time of exposure, reaching higher drug concentrations in whole cells and nuclei compared with exposure to free DOX. The in vitro cytotoxic activity obtained with DOX-loaded FTLs was 10-fold greater than that of the nontargeted liposome formulation, but was not improved over that of free DOX despite the higher cellular drug levels obtained with the targeted liposomes in M109R-HiFR cells. However, if M109R-HiFR cells were exposed to drugs in vitro and tested in an in vivo adoptive assay for tumor growth in syngeneic mice along a 5 week time span, FTL DOX was significantly more tumor inhibitory than free DOX. It is suggested that the biological activity of liposomal DOX released inside the cellular compartment is reduced in vitro due to the aggregated state of DOX, resulting from the liposome drug-loading process, and requires a long period of time and/or an in vivo environment for full expression. PMID- 10815921 TI - The tyrosine kinase inhibitor CGP 57148 (ST1 571) induces apoptosis in BCR-ABL positive cells by down-regulating BCL-X. AB - CGP 57148 is a potent inhibitor of the ABL protein tyrosine kinase and a promising new compound for the treatment of a variety of BCR-ABL-positive leukemias. We used this enzyme inhibitor to characterize the biological effects of BCR-ABL in primary cells and two growth factor-dependent BCR-ABL-transfected cell lines. The effect of CGP 57148 on primary cells is dependent on the stage of differentiation. The growth of maturing chronic myeloid leukemia cells is independent of BCR-ABL in the presence of growth factors. However, the proliferation of leukemic immature cobblestone-forming area cells is almost completely blocked after the inhibition of the BCR-ABL kinase. In the BCR-ABL transfected cell lines, M07/ p210 and Ba/F3/p185, CGP 57148 induces apoptosis by releasing cytochrome c, activating caspase 3, and cleavage of PARP. No alteration of the expression level of the apoptosis regulator BCL-2 was observed. In contrast, BCL-X was down-regulated after exposure to CGP 57148. Inhibitors of signal transduction proteins such as PI-3 kinase, mitogen-activated protein/extracellular signal-regulated kinase kinase, and Janus-activated kinase 2 pathways were not capable of a comparable down-regulation of BCL-X. The Fas/Fas ligand system was not involved either in the induction of apoptosis by CGP 57148. We conclude that the inhibition of the BCR-ABL kinase by CGP 57148 (a) preferentially inhibits the growth of immature leukemic precursor cells, (b) efficiently reverts the antiapoptotic effects of BCR-ABL by down-regulation of BCL-X, and (c) is more effective than the inhibition of the downstream signal transduction pathways of PI-3 kinase, mitogen-activated protein/extracellular signal-regulated kinase kinase, and Janus-activated kinase 2. PMID- 10815922 TI - Tumor necrosis factor-alpha-induced apoptosis in prostate cancer cells through inhibition of nuclear factor-kappaB by an IkappaBalpha "super-repressor". AB - Prostate cancer patients experiencing a relapse in disease often express high serum tumor necrosis factor-alpha (TNF-alpha) levels. Many androgen-insensitive prostate cancer cells are TNF-alpha insensitive because of the expression of antiapoptotic genes as part of the nuclear factor-kappaB (NF-kappaB) family of transcription factors. NF-kappaB stimulates gene transcription when expressed in the nucleus; however, in resting cells, this nuclear import is prevented by association with the cytoplasmic inhibitor IkappaBalpha. This cytoplasmic retention of NF-kappaB is uncoupled by many extracellular signals including low levels of TNF-alpha. During normal cell activation, nuclear translocation of NF kappaB is preceded by phosphorylation and degradation of IkappaBalpha. When phosphorylation is blocked, IkappaBalpha remains intact, thereby blocking NF kappaB translocation to the nucleus and subsequent activation of antiapoptotic genes that cause TNF-alpha insensitivity. We tested whether a "super-repressor" of NF-kappaB activity could be transfected into prostate cancer cells and make them TNF-alpha sensitive. PC-3 and LNCaP cells were stimulated with TNF-alpha (10 ng/ml) for 24 h in the presence or absence of the IkappaBalpha "super-repressor" (p6R-IkappaB(S32A + S36A)). NF-kappaB activity was measured by electrophoretic mobility shift assay and the steady state levels of the cytoplasmic IkappaBalpha protein were measured by Western blot. Secretory IL-6 and IL-6 mRNA were measured by ELISA. p6R-IkappaB(S32A + S36A) blocked the stimulation of NF-kappaB activity by TNF-alpha in prostate cancer cells. It also subsequently decreased IL-6 production by TNF-alpha. We conclude that these data demonstrate that inhibition of NF-kappaB selectively sensitizes previously insensitive prostate cancer cells to TNF-alpha. PMID- 10815923 TI - Taxol and discodermolide represent a synergistic drug combination in human carcinoma cell lines. AB - Recently, three natural products have been identified, the epothilones, eleutherobin, and discodermolide, whose mechanism of action is similar to that of Taxol in that they stabilize microtubules and block cells in the mitotic phase of the cell cycle. In this report, we have compared and contrasted the effects of these new agents in Taxol-sensitive and -resistant cell lines. We also have taken advantage of a human lung carcinoma cell line, A549-T12, that was isolated as a Taxol-resistant cell line and found to require low concentrations of Taxol (2-6 nM) for normal cell division. This study then examined the ability of these new compounds to substitute for Taxol in sustaining the growth of A549-T12 cells. Immunofluorescence and flow cytometry have both indicated that the epothilones and eleutherobin, but not discodermolide, can substitute for Taxol in this Taxol dependent cell line. In A549-T12 cells, the presence of Taxol significantly amplified the cytotoxicity of discodermolide, and this phenomenon was not observed in combinations of Taxol with either the epothilones or eleutherobin. Median effect analysis using the combination index method revealed a schedule independent synergistic interaction between Taxol and discodermolide in four human carcinoma cell lines, an effect that was not observed between Taxol and epothilone B. Flow cytometry revealed that concurrent exposure of A549 cells to Taxol and discodermolide at doses that do not induce mitotic arrest caused an increase in the hypodiploid population, thereby indicating that a possible mechanism for the observed synergy is the potentiation of apoptosis. Our results suggest that Taxol and discodermolide may constitute a promising chemotherapeutic combination. PMID- 10815924 TI - Functional and in situ evidence for nitric oxide production driven by CD40-CD40L interactions in graft-versus-leukemia reactivity. AB - In a murine tumor model, complete tumor remission is achievable at even advanced metastasized stages by transfer of immune T cells from donor B10.D2 (H-2d, Mls(b)) into tumor-bearing DBA/2 (H-2d, Mls(a)) mice. We showed previously that this graft-versus-leukemia (GvL) effect is dependent on synergistic interactions of transferred CD4+ and CD8+ T cells with host sialoadhesin (SER)-positive macrophages. We now show that the CD40-CD40L (CD154) interaction is involved in the induction of inducible nitric oxide synthase (iNOS) expression during adoptive immunotherapy (ADI). We demonstrate that during ADI, the level of CD40 expression in the liver becomes significantly augmented in comparison to livers of tumor-bearing, untreated animals. CD40 expression is found mostly on SER+ macrophages and to a lesser extent on dendritic cells (DCs). In GvL animals, more SER+ macrophages express iNOS than untreated animals. iNOS expressing cells are found in close proximity to apoptotic cells, at early time points of the therapy in areas of metastasis, and at late stages around portal veins, where CD4+ and CD8+ T lymphocytes form clusters with SER+ macrophages. Blocking of CD40L in vivo at days 5 and 20, when all iNOS+ cells express CD40, leads to significantly reduced CD40 and iNOS expression as well as to a marked inhibition of the therapeutic effect. These data provide functional and in situ evidence that the increased CD40 and iNOS expression observed during ADI contribute to the eradication of liver metastases and to the clearance of donor lymphocytes from the liver. PMID- 10815925 TI - Generation and purification of CD8+ melan-A-specific cytotoxic T lymphocytes for adoptive transfer in tumor immunotherapy. AB - Tumor antigens that might serve as potential targets for adoptive T-cell therapy have been defined in different tumor entities, especially in malignant melanoma. To generate conditions to induce primary T-cell responses against different HLA A*0201-restricted melanoma peptides and to allow further expansion of peptide specific T cells for adoptive transfer, CD8+-purified T cells from healthy donors were stimulated with Melan-A-pulsed autologous dendritic cells. Dendritic cells were generated in vitro from monocytes with granulocyte macrophage colony stimulating factor, interleukin-4, and transforming growth factor-beta1. After 3 4 weekly stimulation cycles with Melan-A-pulsed DCs, we were able to induce a strong peptide-specific CTL response in vitro. MHC-peptide tetramer staining revealed a frequency of up to 3.5% CD8+/Melan-A+ T cells. Additional antigen independent expansion with anti-CD3/anti-CD28 monoclonal antibodies together with interleukin-2 gave rise to 600-fold expansion of CD8+ CTLs that maintained Melan A specificity and were able to efficiently lyse Melan-A-expressing melanoma cells. To enrich antigen-specific T cells in vitro, we used a recently established technology for analysis and sorting of live cells according to secreted cytokines. In the present study, we demonstrated that Melan-A-specific T cells can be purified by magnetic separation according to secreted IFN-gamma. These cells revealed a very potent monospecific CTL response, even at low E:T ratios, against Melan-A-pulsed and Melan-A-expressing target cells. Altogether, our study demonstrated that we have developed an efficient method for generating large numbers of peptide-specific T cells in vitro that may be used for adoptive T-cell transfer in tumor immunotherapy. PMID- 10815926 TI - Cyclooxygenase-2 inhibitor induces apoptosis and enhances cytotoxicity of various anticancer agents in non-small cell lung cancer cell lines. AB - In recent years, a combination of two demographic phenomena, an increase in the number of older people in the population and an increase in the incidence of lung cancer with age, has made it mandatory to develop therapeutic modalities with less toxicity for the treatment of inoperable elderly patients with lung cancer. Our study shows that a cyclooxygenase (COX)-2 inhibitor, nimesulide, can inhibit proliferation of non-small cell lung cancer cell lines in vitro in a dose dependent manner, in part by inducing apoptosis even at clinically achievable low concentrations. Our observations also suggest that the responsiveness of non small cell lung cancer to COX-2 inhibitors does not require the presence of wild type p53, but may be influenced by the degree of COX-2 expression. In addition, we found that nimesulide, when used in combination at clinically achievable concentrations, reduced the IC50 values of various anticancer agents by up to 77%, although the level of reduction varied considerably. Because our previous studies have indicated a significantly increased COX-2 expression in up to 70% of adenocarcinoma cases, the present findings are of great clinical interest. In conjunction with the recent development of next generation, highly selective COX 2 inhibitors, they can be expected to lead to even greater efficacy of their use as adjuncts to various anticancer agents for the treatment of high-risk patients without compromising their quality of life. PMID- 10815927 TI - Metabolism of irinotecan (CPT-11) by CYP3A4 and CYP3A5 in humans. AB - 7-Ethyl-10[4-(1-piperidino)-1-piperidino] carbonyloxy-camptothecin (CPT-11), a DNA topoisomerase I inhibitor, undergoes several metabolic pathways to generate conjugated and unconjugated derivatives that could be excreted from the body. The objective of this study was to determine the oxidative metabolites of CPT-11 recovered in human urine samples and to identify cytochrome P450 (CYP) involved in their formation. In addition to the already known metabolites of CPT-11 [SN 38, SN-38-G, 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1 piperidino]carbonyloxycamptothecin (APC), and 7-ethyl-10-(4-amino-1-piperidino) carbonyloxycamptothecin (NPC)], we isolated three oxidized metabolites from the urine of two children and two adults given CPT-11. M1 and M2 (molecular weight, 602) were hydroxylated, respectively, on the CPT moiety and on the terminal piperidine ring of CPT-11. M3 had a molecular mass of 602, but its urine concentration in patients was too low to establish its chemical structure by liquid chromatography/mass spectrometry. In vitro incubations with cells expressing CYP2C8, CYP2C9, CYP1A1, CYP1A2, or CYP3A7 did not produce any detectable metabolites. Only CYP3A4 produced both APC and NPC, resulting from the oxidation of the piperidinylpiperidine side chain of CPT-11 along with metabolite M2. The metabolism of CPT-11 by CYP3A5 was markedly different because neither APC or NPC nor M2 was produced, whereas only one new metabolite, M4 (molecular weight, 558), was generated by de-ethylation of the CPT moiety. No previous study has reported the presence of the M4 metabolite. Production of APC, NPC, M2, and M4 was prevented by ketoconazole, a specific CYP3A inhibitor. The parameters of CPT-11 biotransformation into M2 and M4 were examined using cell lines expressing, respectively, with CYP3A4 and CYP3A5, indicating that CPT-11 is preferentially metabolized by CYP3A4. In conclusion, CYP3A plays a major role in the metabolism of CPT-11, with some differences of the metabolic profile exhibited by 3A4 and 3A5. PMID- 10815928 TI - Photodynamic therapy with the phthalocyanine photosensitizer Pc 4 of SW480 human colon cancer xenografts in athymic mice. AB - Photodynamic therapy (PDT) using the silicon phthalocyanine photosensitizer Pc 4 [HOSiPcOSi(CH3)2(CH2)3N-(CH3)2] is an oxidative stress associated with induction of apoptosis in various cell types. We assessed the effectiveness of Pc 4-PDT on SW480 colon cancer xenografts grown in athymic nude mice. Animals bearing xenografts were treated with 1 mg/kg body weight Pc 4 and 48 h later were irradiated with 150 J/cm2 672-nm light from a diode laser delivered at 150 mW/cm2. Biochemical studies were performed in xenografts resected at various time points up to 26 h after Pc 4-PDT treatment, whereas tumor size was evaluated over a 4-week period in parallel experiments. In the tumors resected for biochemical studies, apoptosis was visualized by activation of caspase-9 and caspase-3 and a gradual increase in the cleavage of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) to a maximum of approximately 60% of the total PARP present at approximately 26 h. At that time all Pc 4-PDT-treated tumors had regressed significantly. Two signaling responses that have previously been shown to be associated with Pc 4-PDT-induced apoptosis in cultured cells, p38 mitogen activated protein kinase and p21/WAF1/Cip1, were examined. A marked increase in phosphorylation of p38 was observed within 1 h after Pc 4-PDT without changes in levels of the p38 protein. Levels of p21 were not altered in the xenografts in correspondence with the presence of mutant p53 in SW480 cells. Evaluation of tumor size showed that tumor growth resumed after a delay of 9-15 days. Our results suggest that: (a) Pc 4-PDT is effective in the treatment of SW480 human colon cancer xenografts independent of p53 status; (b) PARP cleavage may be mediated by caspase-9 and caspase-3 activation in the Pc 4-PDT-treated tumors; and (c) p38 phosphorylation may be a trigger of apoptosis in response to PDT in vivo in this tumor model. PMID- 10815929 TI - Antitumor action of physiological estradiol on tamoxifen-stimulated breast tumors grown in athymic mice. AB - The estrogen receptor (ER)-positive MCF-7 breast cancer cell line can be transplanted into athymic mice and grown into tumors with estradiol (E2) support. Tamoxifen (TAM) blocks E2-stimulated tumor growth; however, continuous TAM treatment results in transplantable tumors within a year that will grow with either E2 or TAM (M. M. Gottardis and V. C. Jordan, Cancer Res., 48: 5183-5187, 1988). Although this model may represent the development of TAM resistance for the treatment of advanced breast cancer, no laboratory model exists to study the exposure of breast cancer to 5 years of adjuvant TAM therapy. We have addressed this issue and report the development and characterization of two tumor lines, MCF-7TAM and MT2, which have been serially transplanted into TAM-treated athymic mice for >5 years. The MCF-7TAM tumor rapidly regresses in response to E2 and then about 50% of tumors regrow in response to E2. Interestingly, tumor regression does not occur if TAM treatment is stopped, probably because E2 levels are too low in ovariectomized athymic mice. The development of the antitumor effect of E2 was documented for MT2 tumors over a 1-year period; TAM-stimulated tumor growth was retained, but E2 caused progressively less of a stimulatory effect. Most importantly, E2-stimulated tumors that regrew after initial tumor regression in both MCF-7TAM and MT2 lines were again responsive to TAM to block E2-stimulated growth. Unlike MCF-7 tumors, the MT2 tumor line contains a single point mutation, Asp351Tyr, in the ER, which was retained after the development of E2-stimulated regrowth. The mutation is associated with increased estrogen-like actions for the TAM-ER complex (A. S. Levenson et al., Br. J. Cancer, 77: 1812 1819, 1998), but we conclude that the mutant ER is not required for TAM resistance. On the basis of the new breast cancer models presented, we propose a cyclic sensitivity to TAM that may have important clinical implications: (a) it is possible that a woman's own estrogen may produce an antitumor effect on the presensitized micrometastatic disease after 5 years of TAM. Long-term antitumor action occurs because the drug is stopped, but resistance accumulates and tumors start to grow if adjuvant therapy is continued; and (b) although in the clinic TAM-resistant tumors respond to second-line therapies that cause estrogen withdrawal, e.g., pure antiestrogens or aromatase inhibitors, estrogen therapy may also be effective and return the tumor to TAM responsiveness. In this way, a hormone-responsive tumor may be controlled longer in the patient with advanced disease. PMID- 10815930 TI - Endoglin is a suitable target for efficient imaging of solid tumors: in vivo evidence in a canine mammary carcinoma model. AB - Increasing evidence suggests that endoglin (CD105) is a new powerful marker of neovascularization in solid malignancies; thus, using breast cancer as a model, we investigated whether targeting of CD105 by monoclonal antibody (mAb) MAEND3 can be used for in vivo imaging of solid tumors. Immunohistochemistry and flow cytometry identified differential expression of CD105 on breast cancer and endothelial cells; in fact, neoplastic cells were weakly and rarely stained by mAb MAEND3, which in contrast, strongly and invariably stained blood vessel endothelia within the breast adenocarcinomas investigated and cultured endothelial cells. Moreover, in contrast to CD31, which currently represents the reference marker to assess angiogenetic activity, CD105 expression was highest in semiconfluent and actively proliferating endothelial cells, and it progressively decreased as cells reached tight confluency and low [3H]thymidine uptake. i.v. administration of 18 MBq of 125I-labeled mAb MAEND3 efficiently imaged spontaneous mammary adenocarcinomas in two dogs; the uptake of radiolabeled mAb was rapid and intense because tumor: background ratios of 8.2:1 and 9.3:1 were reached 8 h after mAb administration, in the absence of immediate and/or long term clinical side effects. Altogether, our present data suggest that targeting of CD105 on tumor-associated blood vessels may represent a new strategy for in vivo imaging of solid malignancies, regardless of their histological origin. PMID- 10815931 TI - Lovastatin potentiates antitumor activity and attenuates cardiotoxicity of doxorubicin in three tumor models in mice. AB - Lovastatin, a drug commonly used in the clinic to treat hypercholesterolemia, has previously been reported to exert antitumor effects in rodent tumor models and to strengthen the antitumor effects of immune response modifiers (tumor necrosis factor alpha and IFN-gamma) or chemotherapeutic drugs (cisplatin). In the present report, we show in three murine tumor cell lines (Colon-26 cells, v-Ha-ras transformed NIH-3T3 sarcoma cells, and Lewis lung carcinoma cells) that lovastatin can also effectively potentiate the cytostatic/cytotoxic activity of doxorubicin. In three tumor models (Co-ion-26 cells, v-Ha-ras-transformed NIH-3T3 sarcoma cells, and Lewis lung carcinoma cells) in vivo, we have demonstrated significantly increased sensitivity to the combined treatment with both lovastatin (15 mg/kg for 10 days) and doxorubicin (3 x 2.5 mg/kg; cumulative dose, 7.5 mg/kg) as compared with either agent acting alone. Lovastatin treatment also resulted in a significant reduction of troponin T release by cardiomyocytes in doxorubicin-treated mice. This observation is particularly interesting because lovastatin is known to reduce doxorubicin-induced cardiac injury. PMID- 10815932 TI - Antitumor effect and potentiation of cytotoxic drugs activity in human cancer cells by ZD-1839 (Iressa), an epidermal growth factor receptor-selective tyrosine kinase inhibitor. AB - Transforming growth factor alpha (TGF-alpha) is an autocrine growth factor for human cancer. Overexpression of TGF-alpha and its specific receptor, the epidermal growth factor receptor (EGFR), is associated with aggressive disease and poor prognosis. The EGFR has been proposed as a target for anticancer therapy. Compounds that block ligand-induced EGFR activation have been developed. ZD-1839 (Iressa) is a p.o.-active, quinazoline derivative that selectively inhibits the EGFR tyrosine kinase and is under clinical development in cancer patients. The antiproliferative activity of ZD-1839 alone or in combination with cytotoxic drugs differing in mechanism(s) of action, such as cisplatin, carboplatin, oxaliplatin, paclitaxel, docetaxel, doxorubicin, etoposide, topotecan, and raltitrexed, was evaluated in human ovarian (OVCAR-3), breast (ZR 75-1, MCF-10A ras), and colon cancer (GEO) cells that coexpress EGFR and TGF alpha. ZD-1839 inhibited colony formation in soft agar in a dose-dependent manner in all cancer cell lines. The antiproliferative effect was mainly cytostatic. However, treatment with higher doses resulted in a 2-4-fold increase in apoptosis. A dose-dependent supra-additive increase in growth inhibition was observed when cancer cells were treated with each cytotoxic drug and ZD-1839. The combined treatment markedly enhanced apoptotic cell death induced by single-agent treatment. ZD-1839 treatment of nude mice bearing established human GEO colon cancer xenografts revealed a reversible dose-dependent inhibition of tumor growth because GEO tumors resumed the growth rate of controls at the end of the treatment. In contrast, the combined treatment with a cytotoxic agent, such as topotecan, raltitrexed, or paclitaxel, and ZD-1839 produced tumor growth arrest in all mice. Tumors grew slowly for approximately 4-8 weeks after the end of treatment, when they finally resumed a growth rate similar to controls. GEO tumors reached a size not compatible with normal life in all control mice within 4-6 weeks and in all single agent-treated mice within 6-8 weeks after GEO cell injection. In contrast, 50% of mice treated with ZD-1839 plus topotecan, raltitrexed, or paclitaxel were still alive 10, 12, and 15 weeks after cancer cell injection, respectively. These results demonstrate the antitumor effect of this EGFR-selective tyrosine kinase inhibitor and provide a rationale for its clinical evaluation in combination with cytotoxic drugs. PMID- 10815933 TI - Z-Phe-Gly-NHO-Bz, an inhibitor of cysteine cathepsins, induces apoptosis in human cancer cells. AB - An increasing number of studies indicate that cysteine cathepsins contribute to cancer progression, invasion, and metastasis. Here we provide experimental evidence that the cathepsin inhibitor Z-Phe-Gly-NHO-Bz induces rapid apoptotic death in human cancer cell lines. Notably, the Z-Phe-Gly-NHO-Bz-induced apoptosis exhibited independence of p53, caspases, and mitogen-activated protein (MAP) kinases. Taken together, our results prompt the hypothesis that cysteine cathepsin(s) is a universal survival factor for cancer cells, and its inhibition leads to cancer cell apoptosis. The exquisite sensitivity of human cancer cells to CATI-1 indicates that this compound and its derivatives may provide the basis for new treatment programs against a broad spectrum of malignancies. PMID- 10815934 TI - Oral efficacy and bioavailability of a novel taxane. AB - A novel taxane (IDN 5109), originally selected for its ability to overcome P glycoprotein-mediated drug resistance, is characterized by an improved preclinical profile in terms of efficacy and tolerability. Because P-glycoprotein may critically influence intestinal absorption and oral bioavailability of taxanes, the purpose of the study was to evaluate the bioavailability, the pharmacokinetic behavior, and the antitumor activity of the new taxane after oral administration. A comparative study of antitumor activity of Taxol and IDN 5109 given orally was performed in a human breast carcinoma model, MX-1, which is highly responsive to i.v. treatment with both of the taxanes. In contrast to Taxol, which was completely ineffective after administration to MX-1-bearing mice, oral IDN 5109 exhibited an activity comparable with that of i.v. treatment (ie., 100% cures). Again, the maximal tolerated doses were comparable (90 mg/kg, every 4 days for four doses) after i.v. and oral treatment. Three other tumor models (LoVo, IGROV/DDP, and U87) with a variable sensitivity to the drug were used to compare the antitumor effects of i.v. and oral treatment with IDN 5109. The efficacy after oral administration was only slightly lower than that found after i.v. treatment at equivalent doses; but optimal effects were comparable likely as a consequence of the long (>6 h) terminal half-life of oral IDN 5109. The bioavailability of IDN 5109 assessed by comparing area-under-the-curve values after oral and i.v. administrations was approximately 50%. The oral efficacy of the novel taxane, likely related to the inability of the P-glycoprotein to recognize the drug, which allowed an adequate intestinal absorption, is a unique feature among the taxanes and may represent a pharmacological breakthrough in their clinical use. PMID- 10815935 TI - Distinctive potentiating effects of cisplatin and/or ifosfamide combined with etoposide in human small cell lung carcinoma xenografts. AB - Combined modalities are currently used for cancer therapy, although their mechanisms of activity remain incompletely deciphered. The design of new drug combinations suffers from our inability to anticipate accurately their efficacy or toxicity. They can be evaluated in vivo, using human tumors grafted into immunodeficient mice, as we did here with combined protocols used in the clinical setting. Xenografts of small cell lung carcinoma (SCLC) from eight patients were used to test the tumor sensitivity to etoposide (VP16; 12-16 mg/kg/days, days 1, 2, and 3), cisplatin (CDDP; 6-9 mg/kg/day, day 1) and ifosfamide (IFO; 90-210 mg/kg/day, days 1, 2, and 3) as single agents and to evaluate the efficacy of the two-drug or three-drug combinations. Five xenografts came from untreated patients (SCLC-61, SCLC-6, SCLC-10, SCLC-41, and SCLC-96) and three after treatment (SCLC 74, SCLC-101, and SCLC-108). p53 was inactivated in all of them. Tumor growth inhibition, growth delay, and the survival rate of tumor-bearing mice reflected individual SCLC chemosensitivity. As single agents, IFO inhibited tumor growth in a dose-dependent manner, whereas CDDP and VP16 had little or no effect. Both CDDP and IFO potentiated VP16, inducing complete regressions in the most sensitive SCLCs; VP16-IFO was more effective than VP16-CDDP, with complete regressions in six versus three of the eight tumors tested, respectively. CDDP-IFO was less effective than VP16-IFO, with three of eight SCLCs giving complete regressions. The three-drug combination led to modest improvement over the best two-drug combination but only for sensitive SCLCs. Because drug-responses distinguished two classes of SCLCs, as sensitive or refractory, MDR1, glutathione S-transferase pi, lung-related multidrug resistance protein, multidrug resistance protein, and topoisomerase IIalpha mRNA expression was studied by semiquantitative reverse transcription. There was no correlation with SCLC sensitivity; topoisomerase IIalpha and multidrug resistance protein was expressed in all cases, lung-related multidrug resistance protein and glutathione S-transferase pie in seven of eight, and MDR1 gene in four of eight. In conclusion, these SCLC xenografts displayed a pattern of chemotherapy response close to that observed in patients. This model confirmed that in two-drug combinations, each component potentiated the effects of the other, with VP16-IFO tending to be the best two-drug combination, both of which were more effective than VP16-CDDP and better tolerated than CDDP-IFO. The addition of a third agent gave a modest, if any, therapeutic benefit in the responders but none in refractory SCLCs. There was no correlation between the extent of response and resistance markers. PMID- 10815936 TI - Schedule-dependent antagonism of gemcitabine and cisplatin in human anaplastic thyroid cancer cell lines. AB - Anaplastic thyroid carcinoma (ATC) affects primarily elderly patients, with a median survival of 4-12 months after diagnosis. Presently, under clinical investigation the combination of cisplatin (CDDP) and gemcitabine (GEM) has promising activity in several of human tumor types. To develop new approaches for therapy of ATC, we evaluated the antineoplastic activity of GEM and CDDP alone (1 h and 24-h drug exposure) or in combination in the ATC cell lines SW1736, 8505C, C643, and HTh74. IC50 values were determined by the sulforhodamine B assay, activity was evaluated by the relative antitumor activity (RAA) and drug interaction assessed by isobologram analysis. GEM seemed to be active in ATC, with RAA ranging from 12-114 and CDDP only modestly active (RAA, 0.24-1.4). In four different drug schedules tested, the drug combination was additive when GEM preceded CDDP exposure (combination index, approximately 1), whereas when CDDP preceded GEM exposure the combination was significantly antagonistic (combination index, >1). In SW1736 and 8505C cells, we observed a strong S phase arrest and DNA synthesis inhibition 24 h after a 1-h exposure to an IC50 of CDDP. On the basis of molecular drug targets, cell cycle arrest points, and DNA synthesis inhibition, a model was developed to explain the interaction observed for the combination of GEM and CDDP. In conclusion, GEM shows promising cytostatic activity in ATC. Interaction of GEM and CDDP was schedule and dose dependent, favoring a regime in which GEM is followed by CDDP. Additionally, our model system suggests that DNA-synthesis inhibition and S phase arrest may be important determinants for the drug interaction between GEM and CDDP. PMID- 10815937 TI - Antitumor and antimetastatic activity of ribozymes targeting the messenger RNA of vascular endothelial growth factor receptors. AB - Chemically stabilized hammerhead ribozymes are nuclease-resistant, RNA-based oligonucleotides that selectively bind and cleave specific target RNAs. Due to their potential for specifically inhibiting gene expression, ribozymes are being investigated for therapeutic applications as well as for the elucidation of gene function. In particular, we have investigated ribozymes that target the mRNA of the vascular endothelial growth factor (VEGF) receptors because VEGF signaling is an important mediator of tumor angiogenesis and metastasis. Here we report pharmacodynamic studies testing anti-Flt-1 (VEGFR-1) and anti-KDR (VEGFR-2) ribozymes in animal models of solid tumor growth and metastasis. Ribozymes targeting either Flt-1 or KDR significantly inhibited primary tumor growth in a highly metastatic variant of Lewis lung carcinoma. However, only treatment with the anti-Flt-1 ribozyme resulted in a statistically significant and dose dependent inhibition of lung metastasis in this model. The anti-Flt-1 ribozyme was then tested in a xenograft model of human metastatic colorectal cancer in which significant inhibition of liver metastasis was observed. Taken together, these data represent the first demonstration that synthetic ribozymes targeting VEGF receptor mRNA reduced the growth and metastasis of solid tumors in vivo. PMID- 10815939 TI - The science of instrumentation and measurements PMID- 10815938 TI - Interleukin 8 expression regulates tumorigenicity and metastases in androgen independent prostate cancer. AB - Interleukin 8 (IL-8) is mitogenic and chemotactic for endothelial cells. Within a neoplasm, IL-8 is secreted by inflammatory and neoplastic cells. The highly metastatic PC-3M-LN4 cell line overexpresses IL-8 relative to the poorly metastatic PC-3P cell line. We evaluated whether IL-8 expression by human prostate cancer growing within the prostate of athymic nude mice regulates tumor angiogenesis, growth, and metastasis. PC-3P cells were transfected with the full length sense IL-8 cDNA, whereas PC-3M-LN4 cells were transfected with the full sequence antisense IL-8 cDNA. Control cells were transfected with the neomycin resistance gene (Neo). In vitro, sense-transfected PC-3P cells overexpressed IL-8 specific mRNA and protein, which resulted in up-regulation of matrix metalloproteinase 9 (MMP-9) mRNA, and collagenase activity, resulting in increased invasion through Matrigel. After antisense transfection of the PC-3M LN4 cells, IL-8 and MMP-9 expression, collagenase activity, and invasion were markedly reduced relative to controls. After orthotopic implantation, the sense transfected PC-3P cells were highly tumorigenic and metastatic, with significantly increased neovascularity and IL-8 expression compared with either PC-3P cells or controls. Antisense transfection significantly reduced the expression of IL-8 and MMP-9 and tumor-induced neovascularity, resulting in inhibition of tumorigenicity and metastasis. These results demonstrate that IL-8 expression regulates angiogenesis in prostate cancer, in part by induction of MMP 9 expression, and subsequently regulates the growth and metastasis of human prostate cancer. PMID- 10815940 TI - Out-of-this-world MS PMID- 10815941 TI - Automated nitrate monitor PMID- 10815942 TI - MRI for gene expression. PMID- 10815943 TI - Imprinting plastic microchips PMID- 10815944 TI - Top 10 articles. Analytical chemistry looks at 10 seminal papers PMID- 10815945 TI - Lab-on-a-chip: a revolution in biological and medical sciences PMID- 10815946 TI - Microfabricated potentiometric electrodes and their in vivo applications. PMID- 10815947 TI - Voltammetry retrospective PMID- 10815948 TI - Electrochemical detectors for liquid chromatography. PMID- 10815949 TI - Practical tips for cooperative learning PMID- 10815950 TI - High-speed fluorescence detection of explosives-like vapors AB - In this paper, we report on the preparation of novel cross-reactive optical microsensors for high-speed detection of low-level explosives and explosives-like vapors. Porous silica microspheres with an incorporated environmentally sensitive fluorescent dye are employed in high-density sensor arrays to monitor fluorescence changes during nitroaromatic compound (NAC) vapor exposure. The porous silica-based sensor materials have good adsorption characteristics, high surface areas, and surface functionality to help maximize analyte-dye interactions. These interactions occur immediately upon vapor exposure, i.e., in less than 200 ms and are monitored with a high-speed charge-coupled device camera to produce characteristic and reproducible vapor response profiles for individual sensors within an array. Employing thousands of identical microsensors permits sensor responses to be combined, which significantly reduces sensor noise and enhances detection limits. Normalized response profiles for 1,3-dinitrobenzene (1,3-DNB) are independent of analyte concentration, analyte exposure time, or sensor age for an array of one sensor type. Explosives-like NACs such as 2,4 dinitrotoluene and DNB are detected at low part-per-billion levels in seconds. Sensor-analyte profiles of some sensor types are more sensitive to low-level NAC vapor even when in a higher organic vapor background. We show that single-element arrays permit the detection of low-level nitroaromatic compound vapors because of sensor-to-sensor reproducibility and signal averaging. PMID- 10815951 TI - Characterization of methanol-water and acetonitrile-water association using multivariate curve resolution methods AB - The solvatochromic comparison method has been used to probe the interactions of solutes with binary solvent mixtures of methanol-water and acetonitrile-water. The solute spectra recorded in these mixtures are composed of the additive spectral contributions of the different solvated species of the solute, i.e., the water-solvated species, the cosolvent-solvated species, and the species solvated by water-solvent complexes. Multivariate curve resolution-alternating least squares has been used to model the solvation of the solutes as a function of the composition of the binary solvent mixture. Spectra and concentration profiles of the dye surrounded by the different solvation environments have been isolated. For the first time, solute spectra solvated exclusively by methanol-water and acetonitrile-water complexes have been obtained, and the solvatochromic parameters of dipolarity/polarizability and hydrogen-bonding acidity have been estimated for these complex species. PMID- 10815952 TI - Determination of biophysical parameters of polypeptide retro-inverso isomers and their analogues by capillary electrophoresis. AB - The relationship between the electrophoretic mobility, microobs, Stokes radius, rs, ionization state, and solution conformation of the all L-alpha-polypeptide, 1, the corresponding retro-all D-alpha-polypeptide, 2, and several truncated analogues, 3-5, has been investigated under low pH buffer conditions by high performance capillary zonal electrophoresis (HPCZE) with coated capillaries. The results confirm that, under these conditions, the all L-alpha-polypeptide, 1, and its retro-inverso isomer, 2, exhibit nonidentical electrophoretic mobilities and thus different Stokes radii. At higher pH values, i.e., pH 5.0, the electrophoretic behavior of this retro-inverso isomer pair, however, converges. These results indicate that variations in the dipole characteristics of the polypeptide main chain and subtle differences introduced by the spatial constraints of the L-alpha-Pro-->D-alpha-Pro residue replacement lead to differences in the Stokes radii and electrophoretic mobilities of these polypeptides. Since the observed electrophoretic mobilities, microobs, reflect the mean of the mobilities of each charge species participating according to their Stokes radius or their intrinsic charge and mole fraction abundances, the results confirm that polypeptide retro-inverso isomers with unmodified amino and carboxy termini are resolvable. This outcome was achieved despite their notional topographical and conformational similarities as assessed from high-field proton nuclear magnetic resonance (1H NMR) spectroscopy and circular dichroism (CD) spectroscopy. PMID- 10815953 TI - Determination of the ionization state of 11-thioundecyl-1-phosphonic acid in self assembled monolayers by chemical force microscopy AB - The present article describes the preparation and preliminary characterization of a novel phosphate-functionalized self-assembled monolayer (SAM) and the determination of the surface ionization states of the phosphate headgroup in aqueous solutions by chemical force microscopy (CFM). The phosphate headgroup used was PO(OH)2, a diprotic acid. The adhesion force between an AFM probe and a flat substrate, both of which were chemically modified with the same phosphate SAM, was also measured as a function of pH and ionic strength. At low ionic strength (10(-4) M), two peaks were observed in the force titration curve (adhesion force versus pH) at pH 4.5 and 8.4. The two peaks are positioned 2.4 and 1.2 pH units higher, respectively, than the acid dissociation constants obtained for the phosphate group free in aqueous solution. At high ionic strength (10(-1) M), the adhesion forces were reduced by 1 order of magnitude and the peaks were replaced by shoulders similar to those previously reported for acid force titrations. On the basis of JKR theory, the surface pKa values of the phosphate group in high ionic strength solutions were found to be 4.5 and 7.7, respectively. However, in light of the effects of ionic strength on the force titration curves, we discuss the applicability of JKR theory to nanoscopic measurements of adhesion force and surface pKa. PMID- 10815954 TI - Probing specific sequences on single DNA molecules with bioconjugated fluorescent nanoparticles. AB - Nanometer-sized fluorescent particles (latex nanobeads) have been covalently linked to DNA binding proteins to probe specific sequences on stretched single DNA molecules. In comparison with single organic fluorophores, these nanoparticle probes are brighter, are more stable against photobleaching, and do not suffer from intermittent on/off light emission (blinking). Specifically, we demonstrate that the site-specific restriction enzyme EcoRI can be conjugated to 20-nm fluorescent nanoparticles and that the resulting nanoconjugates display DNA binding and cleavage activities of the native enzyme. In the absence of cofactor magnesium ions, the EcoRI conjugates bind to specific sequences on double stranded DNA but do not initiate enzymatic cutting. For single DNA molecules that are stretched and immobilized on a solid surface, nanoparticles bound at specific sites can be directly visualized by multicolor fluorescence microscopy. Direct observation of site-specific probes on single DNA molecules opens new possibilities in optical gene mapping and in the fundamental study of DNA-protein interactions. PMID- 10815955 TI - Finite element simulation of an electroosmotic-driven flow division at a T junction of microscale dimensions AB - A finite element formulation is developed for the simulation of an electroosmotic flow in rectangular microscale channel networks. The distribution of the flow at a decoupling T-junction is investigated from a hydrodynamic standpoint in the case of a pressure-driven and an electroosmotically driven flow. The calculations are carried out in two steps: first solving the potential distribution arising from the external electric field and from the inherent zeta potential. These distributions are then injected in the Navier Stokes equation for the calculation of the velocity profile. The influence of the various parameters such as the zeta potential distribution, the Reynolds number, and the relative channel widths on the flow distribution is investigated. PMID- 10815956 TI - Chelating scintillation fibers for measurements of 137Cs AB - Solid scintillating fibers, coated with a dual-mechanism bifunctional polymer shown to bind Cs(I) in alkaline solutions, were developed for measurement of 137Cs. The effect of the epoxy-polymer coating thickness on attenuation of the signal from 137Cs was evaluated. After optimal coating conditions were determined, both scintillation fiber and resin functions were retained, producing stable field-ready fibers. Temporal studies were performed to examine the kinetics of 137Cs uptake into the resin. Calibration curves of the fiber response were generated by beta-emission from solutions of 137Cs dissolved in 1 M sodium hydroxide. The plots exhibited a linear response over a range of 4-3200 nCi/mL (3.4 x 10(-10)-2.7 x 10(-7) M 137Cs), with a limit of detection of 3.65 nCi/mL (approximately 42 parts per trillion 137Cs). The distribution coefficient of Cs was determined to be 490 +/- 50 mL/g from these measurements. Selectivity studies of the resin were performed in the presence of 100 fold excess of Sr(II), Al(III), and nonradioactive Cs(I). Linear calibration plots were obtained in the presence of these potential interferences, but at a reduced sensitivity. The fibers were also used to evaluate the 137Cs content of a mock tank waste sample to show the potential of the fibers in complicated matrixes. PMID- 10815957 TI - Direct observation of calcium-independent intercellular ATP signaling in astrocytes. AB - Adenosine triphosphate (ATP) is assumed to be involved in the regulation of many extracellular signaling systems including calcium wave propagation. So far all supportive evidence is indirect, such as monitoring changes in intracellular calcium on application of extracellular ATP or off-site measurement of ATP from superfusates. Furthermore, the causal relationships among the various signaling agents are still unclear. A novel chemiluminescence dynamic imaging method was developed to monitor ATP release from living biological cells. The assay has linear response over 3 orders of magnitude for fixed concentrations of enzyme and cofactors, with a correlation coefficient of 0.999. The detectability of ATP is down to 10(-8) M at millisecond exposure times with an intensified charge-coupled device camera. The direct imaging of ATP waves in astrocyte cultures was performed together with Fluo-3-Ca imaging at millisecond temporal resolution and micrometer-scale spatial resolution. We discovered that extracellular ATP mediates intercellular calcium wave propagation, but surprisingly, release and propagation of ATP are not calcium dependent. Therefore, ATP rather than Ca or IP3 is the primary intercellular signaling messenger. PMID- 10815958 TI - Relationships among resonant frequency changes on a coated quartz crystal microbalance, thickness changes, and resistance responses of polymer-carbon black composite chemiresistors AB - The relationships among frequency changes on a film-coated quartz crystal microbalance, thickness changes, and dc resistance changes have been investigated for carbon black-insulating polymer composite vapor detectors. Quartz crystal microbalance (QCM) measurements and ellipsometry measurements have been performed simultaneously on polymer films that do not contain carbon black filler to relate the QCM frequency change and the ellipsometrically determined thickness change to the analyte concentration in the vapor phase. In addition, quartz crystal microbalance measurements and dc resistance measurements on carbon black composites of these same polymers have been performed simultaneously to relate the QCM frequency change and dc electrical resistance response to the analyte concentration in the vapor phase. The data indicate that the dc resistance change is directly relatable to the thickness change of the polymers and that a variety of analytes that produce a given thickness change produce a constant resistance change for each member of the test set of polymers investigated in this work. PMID- 10815959 TI - Electrochemical pretreatment of polycrystalline gold electrodes to produce a reproducible surface roughness for self-assembly: a study in phosphate buffer pH 7.4 AB - It has been emphasized in several studies that the state of the surface, including the surface roughness, is very important for the reproducible formation of high-quality self-assembled monolayers on gold. The pulsed-potential pretreatment procedure described in this paper can, in a reproducible way, reduce the surface roughness of mechanically polished polycrystalline gold electrodes by a factor 2. The developed procedure, in which the gold is alternately oxidized and reduced, has been optimized for use in a flow system (100 mM phosphate buffer pH 7.4). The influence of the pretreatment procedure on the surface roughness of the electrodes has been studied by in-situ oxygen adsorption measurements using cyclic voltammetry. The most effective pulse regime in producing a gold surface with a reproducible and relatively low surface roughness is a triple-potential pulse waveform, with potentials of +1.6, 0.0, and -0.8 Vvs SCE and pulse widths of 100 ms for each potential. Prolonged pulsing for 2000-5000 s with the gold working electrode in a flow-through cell showed an electropolishing effect, i.e., a decrease of the roughness in time. Flow conditions are very important: the roughness decreased faster at higher flow rates, while an increase was observed without flow. A process of reconstruction and dissolution of gold during application of the potential pulses under flow conditions is assumed to account for the observed phenomena. A self-assembled monolayer of thioctic acid with reproducible characteristics, determined with impedance measurements, could be formed on a pretreated gold surface. PMID- 10815960 TI - Multimicrobial sensor using microstructured three-dimensional electrodes based on silicon technology. AB - Two microbial strains with different substrate spectra were immobilized separately within a single biosensor chip featuring four individually addressable platinum electrodes. These were sputtered onto the inner surface of four isolated pyramidal cavities ("containments") micromachined on a silicon wafer. The biosensor chip was integrated into a flow-through system to measure the oxygen consumption of the immobilized microorganisms in the presence of assimilable analytes. As a model system, a yeast for the determination of biochemical oxygen demand (BOD) and a strain capable of degrading polycyclic aromatic hydrocarbons (PAH) were chosen. It was shown that the simple and mass-producible containment sensor exhibits good performance data: lower detection limit 0.1 mg/L naphthalene and 1 mg/L sensor-BOD; calibration range up to 30 mg/L; precision 3-6%; response time 2-3 min; service life up to 40 days; shelf life at 4 degrees C 6 months. The versatility of the multimicrobial sensor was demonstrated by measuring ordinary municipal wastewater samples as well as various aqueous samples contaminated with PAH. The concept of a multimicrobial sensor not only enlarges the substrate spectrum for sum parameters such as BOD but leads to additional information which allows for a more differentiated and immediate knowledge of sample composition. Using chemometrical data analysis, the multimicrobial sensor lays a foundation for developing an "electronic tongue". PMID- 10815961 TI - Potentiometric detection of carboxylic acids, phosphate esters, and nucleotides in liquid chromatography using anion-selective coated-wire electrodes. AB - An all-solid-state ion-selective membrane electrode incorporating a lipophilic anion exchanger was used in a flow-through potentiometric detector for the LC determination of organic anions of biological interest. Different metabolic intermediates (mono-, di-. and tricarboxylic acids, sugar phosphates, and nucleotides) were detected sensitively after separation on a pellicular anion exchange chromatographic column. The electrode was coated by directly casting the electroactive mixture on a glassy carbon support of 3 mm diameter and used in a wall-jet-type flow cell. The analysis conditions were optimized to obtain both efficient separation and sensitive detection. Calibration curves showed a logarithmic dependence on the injected concentration for concentrations higher than 5.0 x 10(-5) M and a linear dependence for injected concentrations below this value. Under isocratic conditions, detection limits of 5.0 x 10(-7) M (25 pmol) were attained when a sodium hydroxide solution was used as an eluent. No suppressor system was needed in this case. The relative standard deviation for consecutive injections was 0.3% (n = 15), and the electrode lifetime was at least 2 months. The utility of potentiometric detection is further demonstrated in a gradient elution separation for single-run analysis of a synthetic mixture of biochemical compounds containing carboxylic acids, phosphate esters, and nucleotides. PMID- 10815962 TI - An electrochemical impedance measurement technique employing Fourier transform AB - We describe a novel technique for measuring electrochemical impedance, in which the electrode potential is ramped to a desired bias potential and a small potential step is applied to the working electrode after a short time delay. Fourier transform of the first derivative of the current signal thus obtained provides ac currents in the frequency domain, which allows the computation of impedances of the electrode/electrolyte interface in the whole frequency range. A home-built data acquisition system for these measurements and the results obtained therefrom were used for the measurements. The advantage of the technique includes an extremely short time of less than 1 ms for impedance measurements in the whole frequency region while equilibrium conditions of the electrochemical system are being maintained before and after the measurements, among many others. This technique is expected to revolutionize electrochemical measurements and to find important applications such as in situ measurements during battery cycling, corrosion testing, and other electrochemical experiments. PMID- 10815963 TI - A theoretical description of microdialysis with mass transport coupled to chemical events. AB - A random-walk simulation of microdialysis is used to examine how a reaction that consumes analyte in the medium external to the probe affects the extraction and recovery processes. The simulations suggest that such a reaction can promote the extraction process while simultaneously inhibiting the recovery process, which appears to be consistent with recent experimental evidence of asymmetry in the extraction and recovery of the neurotransmitter, dopamine, during brain microdialysis. This suggests that quantitative microdialysis strategies that rely on the extraction fraction as a measure of the probe recovery value, such as the no-net-flux method, will produce an underestimate of the analyte concentration in the external medium when that analyte is consumed by a reaction in the external medium. Furthermore, if experimental conditions arise under which the kinetics of the reaction are changed, then changes in the extraction and recovery processes are likely to occur as well. The implications of these theoretical findings for the quantitative interpretation of in vivo microdialysis results obtained for the neurotransmitter dopamine are examined. PMID- 10815964 TI - Direct potentiometric information on total ionic concentrations. AB - Polymeric membrane ion-selective electrodes exhibit an apparently super-Nernstian response at low sample activities if inner solutions are used that induce strong zero-current fluxes of primary ions toward the inner compartment. This is due to the limited ion fluxes in the aqueous boundary layer near the membrane. In the presence of labile complexes, the effective flux rate is increased and the emf depends on the total concentration of the ions. The concept is illustrated experimentally with calcium-selective electrodes based on the ionophore N,N dicyclohexyl-N',N'-dioctadecyl-3-oxapentanediamide (ETH 5234) that either respond to total or free ion concentrations. Samples can be distinguished that contain varying levels of total calcium but are all buffered with EDTA to the same free calcium concentration of 5 x 10(-8) M. PMID- 10815965 TI - Immobilized parathion hydrolase: an amperometric sensor for parathion. AB - An amperometric enzyme biosensor for the direct measurement of parathion was developed. The biosensor is based on parathion hydrolase from Pseudomonas sp. isolated from contaminated soil. The enzyme, which was immobilized on a carbon electrode, catalyzes the hydrolysis of parathion to form p-nitrophenol, which was detected by its anodic oxidation. The enzymatic and electrochemical reactions were examined and optimized. Screen-printed electrodes and a microflow injection system provide the means to significantly reduce the volume of the detected samples. Pulsed techniques further increased the sensitivity of the measurement. The current signal was linearly related to the parathion concentration, and the detection limit was less than 1 ng/mL. The biosensor is rapid as well and can be used outdoors and indoors by a nonqualified person. PMID- 10815967 TI - Electrochemically modulated preconcentration and matrix elimination for organic analytes coupled on-line with electrospray mass spectrometry. AB - Demonstrated for the first time is the use of electrochemically modulated preconcentration and sample matrix elimination combined on-line with electrospray mass spectrometry (EMPM/ES-MS) for the enhanced analysis of organics by ES-MS. EMPM is similar to adsorptive stripping analysis. Accumulation of the targeted analytes at the working electrode of an on-line electrochemical flow cell is accomplished via a nonelectrolytic adsorption process that is controlled through the proper combination of the solvent system, the working electrode material, and applied potential. Once on the electrode, the analyte may be washed free of sample matrix components detrimental to mass spectrometric detection. The potential applied to the electrode during the detection step is chosen to release or strip the analytes unaltered back into the solvent stream for mass spectrometric detection rather than to oxidize or reduce them as would be the case for electrochemical detection. Thus, retention and elution of a target analyte with EMPM are controlled by switching the working electrode potential, rather than via a switch in mobile-phase composition, as is done in more traditional preconcentration and cleanup schemes used on-line with ES-MS. The proof-of-principle studies described here use the breast cancer drug tamoxifen and a metabolite, 4-hydroxytamoxifen, as the target analytes. A thin-layer, flow by electrode cell with a glassy carbon working electrode is used as the preconcentration device. The nature of the working electrode, the solvent systems, and the electrode potentials necessary to accumulate and strip tamoxifen and 4-hydroxytamoxifen are discussed. Calibration curves were fitted using the Langmuir isotherm. Detection limits (DLs) using a 5.0 min preconcentration period with selected reaction monitoring for tamoxifen (m/z 372 --> 72) were bracketed as 0.010 nM < DL < 0.025 nM. The ability to simultaneously detect low nanomolar levels of both tamoxifen and 4-hydroxytamoxifen in pristine solution and 1/10 diluted urine is also demonstrated. PMID- 10815966 TI - Detection of chemically induced DNA damage by derivative square wave voltammetry. AB - Damage of DNA films after reaction with styrene oxide was detected using derivative square wave voltammetry. Double-stranded (ds) DNA films with initially low backgrounds developed oxidation peaks for DNA bases during incubation with styrene oxide. Films were prepared on pyrolytic graphite (PG) electrodes by casting mixtures of DNA with the poly(ester sulfonic acid) ionomer Eastman AQ38S or by covalent binding of DNA onto oxidized PG. While both types of films gave oxidation peaks in the region 0.6-1.1 V vs SCE after incubations with styrene oxide, DNA/AQ films gave the best signal-to-background ratios. Damage of DNA by reaction with styrene oxide under the electrode incubation conditions was confirmed by capillary electrophoresis. Total integrals of oxidation peaks increased with time of incubation with styrene oxide. Relative peak heights depended on the type of DNA in the order calf thymus ds DNA > salmon sperm ds DNA > supercoiled ds DNA > highly polymerized calf thymus ds DNA. PMID- 10815968 TI - Catalyzed hydrolysis of nerve gases by metal chelate compounds and potentiometric detection of the byproducts. AB - A new method for detection of nerve gases, Sarin and Soman, was proposed on the basis of their catalyzed hydrolysis by metal chelate compounds and potentiometric detection of the byproduct, fluoride ion. Diisopropyl fluorophosphate (DFP) was used as a nerve gas mimic. The copper ion chelates shift the potential of the fluoride ion-selective electrode to more positive stable potential, which is beneficial for lowering the detection limit. In the presence of DFP, the electrode potential decreases rapidly with time due to the catalyzed hydrolysis of DFP and the production of fluoride ion. This method is sensitive, selective, and reproducible. The detection limit for DFP is 2 x 10(-6) M with a potential drop between 40 and 60 mV. PMID- 10815969 TI - Nonaqueous capillary electrophoresis of fatty acids derivatized with a near infrared fluorophore. AB - Saturated linear fatty acids, derivatized with a near-infrared absorbing fluorescent dye, were separated in 100% methanol with 12.5 mM tetraethylammonium chloride added as a charge carrier. Separation at 380 V/cm was acceptable for acids that differed in length by a single carbon. The labeled linear fatty acids behaved as random coils in the nonaqueous separation medium, as shown in a fit to a simple theoretical expression. However, even in 100% methanol with a trimethylsilylated capillary, significant adsorption to the capillary wall occurred, which reduced resolution and slowed the separation. Addition of water to the methanol medium caused significant differences in separation behavior of high molecular weight acids (>C16). Addition of a cetyltrimethylammonium bromide surfactant to the separation medium dynamically coated the capillary and greatly improved the separation. The surfactant also interacted with the acyl tail, apparently causing it to collapse. Resolution in an optimal separation medium (20 mM surfactant) ranged from 1.6 to 1.1, depending on chain length, and theoretical plate heights were under 4 microm (N > 10(5)). Resolution was more than adequate to separate stearic (C18:0) from oleic (C18:1) acid, as well as other unsaturated C18 homologues. PMID- 10815970 TI - Analysis of solids by laser ablation and resonance-enhanced laser-induced plasma spectroscopy AB - An analytical technique based on resonance-enhanced laser-induced plasma spectroscopy was demonstrated. Pellets of potassium iodate containing trace amounts of sodium were ablated by a 532-nm laser pulse in air. After 30 ns, the plasma plume was intercepted by a 404.4-nm laser pulse to resonantly photoionize the potassium atoms in the vapor plume. The 589-nm emissions of the sodium were found to be significantly enhanced. The enhancement was shown to depend critically on the profile of the 532-nm beam as well as the spatial overlap of the two laser pulses. Using this double-pulse scheme, the mass detection limit for sodium was estimated to be about 200 pg, which was five times better than that obtained by using the 532-nm laser pulse alone. PMID- 10815971 TI - Raman spectroscopy as a discovery tool in carbohydrate chemistry. AB - Raman spectra of nine anomerically stable monosaccharides have been obtained in aqueous solution in the 700-1700 cm(-1) spectral range. Good-quality spectra are obtained of solutions with concentrations as low as 10 mM and volumes as small as 15 microL. Interestingly, the Raman spectra appear to be exquisitely sensitive to the configuration of the carbon centers; unique spectra are obtained of all nine monosaccharides. The unique Raman spectral fingerprint observed for each monosaccharide, and for each anomer of each monosaccharide, suggests that Raman spectroscopy may be a useful technique for the identification and characterization of biologically relevant oligosaccharides. To test this idea, Raman spectra of three unknown disaccharides were obtained in a single-blind study. Identification of the individual monosaccharide components and their anomeric configuration was completely successful. All of these results suggest that development of Raman spectroscopy as a fast, sensitive discovery tool in glycobiology and carbohydrate chemistry is straightforward. PMID- 10815972 TI - Quantitative analysis of film coating in a fluidized bed process by in-line NIR spectrometry and multivariate batch calibration. AB - A method is described which enables real-time analysis of film coating on pharmaceutical pellets during an industrial manufacturing process. Measurements were conducted on the solid particulate material by near-infrared (NIR) spectrometry utilizing a diffuse reflectance fiber-optic probe positioned inside a fluidized bed process vessel. Time series of NIR spectra from 11 batches generated a three-way data matrix that was unfolded and modeled by partial least squares (PLS) in a multivariate batch calibration. The process conditions were deliberately varied according to an experimental design. This yielded good predictability of the coating thickness with a best model fit, R2 = 0.97, for one PLS-projection, and a root-mean-square error of calibration = 2.2 microm (range tested 0-50 microm). The regression vector was shown to be highly influenced by responses that are both direct (aliphatic C-H stretch overtones) and indirect (aromatic C-H stretch overtones), from film component and core material, respectively. The impact of different data pre-treatment methods on the normalization of the regression vector is reported. Justification of the process calibration approach is emphasized by good correlation between values predicted from NIR data and reference image analysis data on dissected pellets and a theoretical nonlinear coating thickness growth model. General aspects of in-line NIR on solids and multivariate batch calibration are discussed. PMID- 10815973 TI - On-line time-resolved laser-induced fluorescence of UO2(NO3)2.2TBP in supercritical fluid CO2 AB - Time-resolved laser-induced fluorescence has been shown to be effective for on line measurement of uranyl chelates in supercritical carbon dioxide. Quenching and other factors affecting the method's accuracy were identified and quantified to allow for accurate measurements under variable supercritical fluid conditions. The method was applied to the kinetic analysis of uranium extraction from acidic solutions with tributyl phosphate modified supercritical carbon dioxide. PMID- 10815974 TI - Forbidden light detection from single molecules AB - We present a new concept for ultrasensitive detection of surface-generated fluorescence which is made possible by a new optical module. The detection method leads to an enhancement in fluorescence collection efficiency to more than 65% of the total of emitted light, whereas high-aperture microscope objectives are able to collect 44% at best. Moreover, by employing this new optical module, the detection volume can be restricted to approximately 10(-17) L. This allows for an exceptional discrimination of bulk-generated against surface-generated fluorescence, which may be of great value when surface-binding processes are monitored. We demonstrate the performance of the new detection system by detecting single fluorescent molecules and by determining antigen concentrations down to 5 fmol. PMID- 10815975 TI - Analysis of nitro musk compounds and their amino metabolites in liquid sewage sludges using NMR and mass spectrometry. AB - A method using HRGC ion trap MS/MS for measuring simultaneously amino metabolites and the parent compounds, the nitro musks, an important group of organic fragrance components found in sewage sludges, was developed. The monoamino metabolites were synthesized and characterized by 1H/13C NMR and mass spectroscopy. Among the nitro musks, musk ketone was the major compound, found at an average concentration of 5 microg/kg of dry mass (dm) whereas musk xylene was detected in only one sample (30 microg/kg dm). Three amino metabolites were identified, namely, 1-tert-butyl-3,5-dimethyl-4-amino-2,6-dinitrobenzene, 1,1,3,3,5-pentamethyl-4-nitro-6-aminoindane, and 4-acetyl-1-tert-butyl-3,5 dimethyl-2-nitro-6-aminobenzene, the corresponding reduction products of the nitro musks xylene, moskene, and ketone. These metabolites were present in partly higher concentrations in the sludges than the corresponding nitro musk compounds. Musk xylene and musk moskene were mainly found as their monoamino metabolites, underlining the importance of anaerobic reduction processes in the sewage treatment plant. PMID- 10815976 TI - MALDI quadrupole time-of-flight mass spectrometry: a powerful tool for proteomic research. AB - A MALDI QqTOF mass spectrometer has been used to identify proteins separated by one-dimensional or two-dimensional gel electrophoresis at the femtomole level. The high mass resolution and the high mass accuracy of this instrument in both MS and MS/MS modes allow identification of a protein either by peptide mass fingerprinting of the protein digest or from tandem mass spectra acquired by collision-induced dissociation of individual peptide precursors. A peptide mass map of the digest and tandem mass spectra of multiple peptide precursor ions can be acquired from the same sample in the course of a single experiment. Database searching and acquisition of MS and MS/MS spectra can be combined in an interactive fashion, increasing the information value of the analytical data. The approach has demonstrated its usefulness in the comprehensive characterization of protein in-gel digests, in the dissection of complex protein mixtures, and in sequencing of a low molecular weight integral membrane protein. Proteins can be identified in all types of sequence databases, including an EST database. Thus, MALDI QqTOF mass spectrometry promises to have remarkable potential for advancing proteomic research. PMID- 10815977 TI - Laser desorption/in situ chemical ionization aerosol mass spectrometry for monitoring tributyl phosphate on the surface of environmental particles. AB - The possibility of using real-time aerosol mass spectrometry (RTAMS) for the detection of surface-adsorbed tributyl phosphate (TBP) as an alkali metal adduct has been investigated. Environmental particles contain variable amounts of easily ionizable alkali metals. During laser desorption of surface-adsorbed TBP molecules, Na+ and K+ ions are generated by the interaction of the laser radiation with the particle's material. The alkali metal ions serve as in situ chemical ionization reagents of the neutral analyte molecules. The effect of laser fluence on the signal intensities of the potassium ion and cationized TBP was also studied. The best performance of the instrument was observed with laser fluences that produce high abundances of K+ but low abundances of ions from the particle's bulk material. The relatively low laser fluence, necessary to produce potassium ions, prevents the excessive fragmentation of the analyte. The instrument is capable of real-time monitoring of submonolayer coverage of TBP on the surface of micron-sized particles. PMID- 10815978 TI - Identification of protein vaccine candidates from Helicobacter pylori using a preparative two-dimensional electrophoretic procedure and mass spectrometry. AB - Helicobacter pylori is an important human gastric pathogen for which the entire genome sequence is known. This microorganism displays a uniquely complex pattern of binding to complex carbohydrates presented on host mucosal surfaces and other tissues, through adhesion molecules (adhesins) on the microbial cell surface. Adhesins and other membrane-associated proteins are important targets for vaccine development. The identification and characterization of cell-surface proteins expressed by H. pylori is a prerequisite for the development of vaccines designed to interfere with bacterial colonization of host tissues. However, identification of membrane proteins is difficult using a traditional proteomics approach employing 2D-PAGE. We have used a novel approach in the identification of microbial proteins that employs a rapid preparative two-dimensional electrophoretic separation followed by mass spectrometry and database searches. No pre-enrichment of bacterial membranes was required. The entire process, from sample preparation to protein identification, can be completed in less than 18 hours, and the presence of proteins can be monitored after both the first- and second-dimensional separations using mass spectrometry. We were able to identify 40 proteins from a detergent-solubilized H. pylori preparation; over one-third of these were membrane or membrane-associated proteins. A functionally characterized low-abundance membrane protein, the Leb-binding adhesin, was found in this group. The use of this rapid 2D electrophoretic separation in proteomic studies of H. pylori is expected to speed up the identification of expressed virulence proteins and vaccine targets in this and other microbial pathogens. PMID- 10815979 TI - High-efficiency capillary isoelectric focusing of peptides. AB - Several approaches are presently being developed for global proteome characterization that are based upon the analysis of polypeptide mixtures resulting from digestion of (often complex) mixtures of proteins. Improved methods for peptide analysis are needed that provide for sample concentration, higher resolution separations, and direct compatibility with mass spectrometry. In this work, methods for the high-efficiency capillary isoelectric focusing (CIEF) separation of peptides have been developed that provide for simultaneous sample concentration and separation according to peptide isoelectric point. Under typical nondenaturing CIEF conditions, peptides are concentrated approximately 500-fold, and peptides present at < 1 ng/ microL were detectable using conventional UV detection. CIEF separations of peptides provided much faster measurements of isoelectric points compared with conventional isoelectric focusing in gels. Very small differences in peptide isoelectric points (deltapI approximately 0.01) could be resolved, High-efficiency CIEF separations for complex peptide mixtures from tryptic digestion of yeast cytosol fractions were obtained and showed significant improvement over those obtained using capillary zone electrophoresis and packed capillary reversed-phase liquid chromatography. PMID- 10815980 TI - The possibility of generating high-speed shear-driven flows and their potential application in liquid chromatography AB - An experimental proof of principle is presented for the possibility to use a shear force field to generate a stable, chromatography enabling fluid flow through micrometer and submicrometer channels without the need for a pressure or a voltage gradient. In our setup, we were able to successfully move a color tracer plug at speeds exceeding 2 cm/s through a 0.125-microm-thick and 4-mm-wide channel, without creating a pressure drop or a pressure buildup. By showing that the speed of microchannel flows can be drastically increased by simply switching from one driving force to another, the presented experiments open the road to the development of a new type of chromatography, referred to as shear-driven chromatography, potentially offering unprecedented separation speeds and resolutions and complying perfectly with the present trend toward the miniaturization and parallelization of analytical separation equipment. PMID- 10815981 TI - Headspace membrane introduction mass spectrometry for trace level analysis of VOCs in soil and other solid matrixes AB - A new MIMS-derived technique, headspace membrane introduction mass spectrometry (HS-MIMS), is described for direct trace level analysis of volatile organic compounds (VOCs) in soil and other dry or wet solid matrixes. A silicone membrane interface is placed about 15 cm from the ion source, and a closed airspace (headspace) is created by connecting a toggle valve to the 1/4 in. tubing that connects the membrane interface to the ion source. For the VOC analysis, the headspace is evacuated and the solid sample vessel is heated to 90 degrees C. The VOCs are rapidly desorbed from the sample, pervaporated through the membrane, and preconcentrated for 4 min in the evacuated headspace. Then, the toggle valve is opened and the trapped VOCs are released into the ion source region of a quadrupole mass spectrometer. By electron ionization and selected-ion monitoring, a relatively sharp and intense peak is obtained and used for quantification. The HS-MIMS analysis shows excellent linearity and reproducibility and detection limits for many VOCs typically of 50-100 ng/kg (ppt). PMID- 10815982 TI - Screening of pentachlorophenol-contaminated wood by thermodesorption sampling and photoacoustic detection. AB - The still-remaining high amounts of pentachlorophenol (PCP), used as wood preservative, in buildings and in waste wood are a potential risk for humans and the environment. To ensure a fast and selective measurement of PCP, a screening tool was developed, which is not only sensitive, but, unlike conventional methods, it requires no added chemicals, is simple, cost-effective, mobile, and capable of on-site operation. The instrument combines light-induced thermodesorption sampling followed by an external cavity diode laser based photoacoustic detector. Measurements on wood samples proved that the system can determine PCP to as low as a concentration of 10 microg/cm2 within minutes without destruction of the sample. The system was calibrated with reference to the concentration of PCP impregnated on the wood surface. It is demonstrated that measurements are not influenced by moisture content of the wood samples. PMID- 10815983 TI - A glucose oxidase electrode based on electropolymerized conducting polymer with polyanion-enzyme conjugated dopant. AB - An enzyme immobilization method has been developed by electropolymerization chemistry of conducting polymer which results in a more effective and reproducible enzyme electrode. As a model system, in this study, glucose oxidase (GOD) was conjugated with a polyanion, poly(2-acrylamido-2-methylpropane sulfonic acid), via a poly(ethylene oxide) spacer to improve the efficiency of enzyme immobilization into a conducting polymer. GOD was successfully conjugated with a high conjugation yield of more than 90%, and its bioactivity was preserved. The resulting polyanion-GOD conjugate was used as a dopant for the electrochemical polymerization of pyrrole. Polypyrrole was effectively deposited on a Pt wire working electrode with the polyanion-GOD conjugate. The enzyme electrode responded to glucose concentrations of up to 20 mM with a sensitivity of 40 nA/mM at an applied potential of 0.4 V within a response time of 30 s. Although the response signal decreased at the low applied potential of 0.3 V, the enzyme electrode showed sensitive response signals of about 16 nA/mM up to 20 mM in glucose concentration. Under the deoxygenated condition, reduced but clear response current signal was obtained. The results show that the current signal response of the enzyme electrode to glucose concentration may be produced by mixed mechanisms. PMID- 10815984 TI - The sugar moiety of Tamm-Horsfall protein is affected by the carbohydrate deficient glycoprotein type I syndrome. A case study. AB - As the sugar moiety of Tamm-Horsfall protein (THP) is affected by many pathological conditions, the aim of this study was to examine the influence of carbohydrate-deficient glycoprotein syndrome (CDG) on THP glycans. THP was isolated from urine of one patient with CDG type I and N-glycan profiling, analysis of monosaccharide content, determination of THP reactivity with specific lectins and with anti-THP antibodies were performed. THP of the CDG patient showed markedly lower amounts of all monosaccharides. Diminished amounts of lactosamine-type chains, galactose and alpha2,3 linked sialic acid were expressed in lower reactivity with PHA-L, DSA and MAA, respectively. These modifications were reflected in altered proportions of tetrasialylated and disialylated oligosaccharide chains. THP of the CDG patient reacted slightly more with anti THP antibodies. Our results indicate that the CDG type I affects the THP sugar moiety and slightly enhances the THP immunoreactivity. PMID- 10815985 TI - Complete enzymic synthesis of the mucin-type sialyl Lewis x epitope, involved in the interaction between PSGL-1 and P-selectin. AB - Sialyl Lewis x (sLe(x)) is an established selectin ligand occurring on N- and O linked glycans. Using a completely enzymic approach starting from p-nitrophenyl N acetyl-alpha-D-galactosaminide (GalNAc(alpha1-pNp as core substrate, the sLe(x) oligosaccharide Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1 6)[Gal(bet a1-3)]GalNAc(alpha1-pNp, representing the O-linked form, was synthesized in an overall yield of 32%. In a first step, Gal(beta1 3)GalNAc(alpha1-pNp was prepared in a yield of 52% using UDP-Gal and an enriched preparation of beta3-galactosyltransferase (EC 2.4.1.122) from rat liver. UDP GlcNAc and a recombinant affinity-purified preparation of core 2 beta6-N acetylglucosaminyltransferase (EC 2.4.1.102) fused to Protein A were used to branch the core 1 structure, affording GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc(alpha1 pNp in a yield of >85%. The core 2 structure was galactosylated using UDP-Gal and purified human milk beta4-galactosyltransferase 1 (EC 2.4.1.38) (yield of >85%), then sialylated using CMP-Neu5Ac and purified recombinant alpha3 sialyltransferase 3 (EC 2.4.99.X) (yield of 87%), and finally fucosylated using GDP-Fuc and recombinant human alpha3-fucosyltransferase 6 (EC 2.4.1.152) produced in Pichia pastoris (yield of 100%). Overall 1.5 micromol of product was prepared. MALDI TOF mass spectra, and 1D and 2D TOCSY and ROESY 1H NMR analysis confirmed the obtained structure. PMID- 10815986 TI - Synthesis of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with N-acetylneuraminic acid. AB - In order to develop glycosylated cytokine, recombinant human IL-1alpha was chemically modified with N-acetylneuraminic acid (NANA). NANA with C9 spacer, 8 (hydrazinocarbonyl)octyl 5-acetamido-3, 5-dideoxy-D-glycero-alpha-D-galacto-2 nonulo-pyranosidonic acid potassium salt (6), was synthesized by glycosylation of C9 spacer, 8-[2-N-(benzyloxycarbonyl)hydrazinocarbonyl]octanol, with methylthio glycoside derivatives of NANA in the presence of molecular sieves 3A and methyl (methylthio)sulfonium trifrate in propionitrile, followed by separation of a and beta anomers with a column chromatography and deprotection. Compound 6 was coupled to IL-1alpha by the acyl azide method. The glycosylated IL-1 was purified by anion-exchange chromatography, and NANA coupled to IL-1 was confirmed by oxidation with NaIO4. Based on the molecular weight average number of carbohydrate molecules introduced per molecule of IL-1alpha was estimated to be 2.9. PMID- 10815987 TI - Allylmalonamide as a bivalent linker: synthesis of biantennary GM3-saccharide- keyhole limpet hemocyanin glycoconjugate and the immune response in mice. AB - A biantennary GM3-saccharide (sialyllactoside) derivative (4) was constructed using allylmalonic acid as a bivalent linker, both carboxylic acids of which were condensed with 3-aminopropyl lactoside (2) prior to enzymatic sialylation with a fusion enzyme. While ozonolysis of its allyl group generated a saccharide having a terminal aldehyde (6), we were unable to couple 6 directly to protein by reductive amination. However, extension of the spacer by means of introducing a maleimide group to 6 through its aldehyde group to give 7 enabled the latter to be successfully coupled to thiolated proteins. The average ratios of saccharide to protein were observed to be 35 in KLH conjugate (13) and 9-12 in HSA conjugates (14 and 15). The antisera obtained by immunizing mice with the biantennary sialyllactoside-KLH conjugate (13) together with MPL adjuvant were analyzed by ELISA. Using several structurally related saccharide-HSA conjugates as screening antigens, it was concluded that anti-sialyllactoside antibodies, both IgG and IgM, were effectively raised. This was further supported by competitive inhibition experiments using lactoside (1), sialyllactoside (8) and biantennary sialyllactoside (4) as inhibitors. PMID- 10815988 TI - Purification and characterization of the hemagglutinin-neuraminidase of Porcine rubulavirus LPMV. AB - The Hemagglutinin-Neuraminidase (HN) from the LPMV strain of Porcine rubulavirus was purified from virions by ultracentrifugation in a continuous 20-60% sucrose gradient and by ion exchange chromatography. The HN is a glycoprotein of 66 kDa constituted by 50.5, 13.3 and 13.6% of non polar, uncharged polar, and charged polar amino acids, respectively. The HN contains 4% of carbohydrates, its glycannic portion is constituted by Man, Gal, GlcNAc, GalNAc, and Neu5Ac in 3:3:4:1:1 molar ratios. The HN possesses hemagglutinating activity in the presence of erythrocytes from several animal species, including human ABO, and treating the erythrocytes with neuraminidase or pronase abolishes this activity. The binding specificity of the purified HN was determined by hapten inhibition assays, indicating that the hemagglutinating activity of the HN is specific for sialic acid and Neu5Acalpha2,3Gal-containing structures. PMID- 10815990 TI - Specific inhibition of an alpha-galactosyltransferase from Trypanosoma brucei by synthetic substrate analogues. AB - Since the alpha-D-galactose-(1-->3)-D-galactose epitope has been identified to be the major target in the process of hyperacute rejection of xenografts transplanted from nonprimate donors to humans, specific inhibitors of alpha galactosyltransferases are of broad interest. Using Trypanosoma brucei, a protozoan parasite causing sleeping sickness and Nagana, we have a very useful model system for the investigation of alpha-galactosyltransferase inhibitors, since the variant surface glycoprotein (VSG) accounts for about 10% of the total cell protein an this parasite expresses many different galactosyltransferases including the one catalysing the formation of the Galalpha1-->3Gal epitope. In order to study inhibition of galactosylation on the VSG from Trypanosoma brucei, we designed, synthesized and tested substrate analogues of trypanosomal alpha galactosyltransferases. Effective inhibitors were a pair of diastereoisomeric UDP galactose analogs, in which the galactose residue is linked to UDP via a methylene bridge rather than an ester linkage. Hence, galactose cannot be transferred to the respective acceptor substrate VSG or the synthetic acceptor substrate Manalpha1-->6Manalpha1S-(CH2)7-CH3, which was previously proven to replace VSG effectively [Smith et al. (1996) J Biol Chem 271:6476-82]. Inhibitors have been prepared starting from 1-formyl galactal. The final condensation was performed using UMP morpholidate leading to a pair of diastereomeric compounds in 39% or 30% yield, respectively. These compounds were tested using alpha galactosyltransferases prepared from T. brucei membranes and lactose synthetase from bovine milk. While the K(M)-value for UDP-galactose was determined as 59 microM on bovine lactose synthetase, the K(I)-values for both inhibitors were 0.3 mM and 1.1 mM respectively, showing that these inhibitors are unable to inhibit enzyme activity significantly. However, using the N-glycan specific alpha galactosyltransferase from trypanosomes, the K(M)-value was determined as 20 microM, while the K(I)-values were 34 microM and 21 microM respectively. Interestingly, other trypanosomal alpha-galactosyltransferases, which modify the GPI membrane anchor, are 2 orders of magnitude less effected by the inhibitor. PMID- 10815989 TI - Characterization of distinct Gal:3-O-sulfotransferase activities in human tumor epithelial cell lines and of calf lymph node GlcNAc : 6-O-sulfotransferase activity. AB - We found earlier in human breast and colon tumors, an augmented level of Gal : 3 O-sulfotransferase activities showing, respectively, an acceptor preference to blood group T-hapten (Group A enzymes) or Galbeta1,4GlcNAc (Group B enzymes) on the mucin Core 2 structure [Chandrasekaran EV, Jain RK, Vig R, and Matta KL (1997) Glycobiology 7: 753-68]. The present study reports these enzyme activities in human tumor cell lines and additional tumor specimens. The human colon tumor epithelial cell lines, akin to their parent tumors, express Group B enzyme activity. The acceptor specificity and kinetic properties, such as divalent metal ion activation and pH dependent activity profile, of the colon cancer line LS180 enzyme activity are identical to those of colon tissue specimens. Consistent with breast tumor specimens, the Group A enzyme activity is present in human breast tumor epithelial cell lines, with some exceptions. The Gal : 3-O sulfotransferases show specific binding to Aleuria aurantia lectin, suggesting the presence of asparagine linked carbohydrate chains containing an inner core alpha1,6-fucosyl residue on these enzymes. Calf lymph nodes contain GlcNAc : 6-O sulfotransferase as well as Group A Gal : 3-O-sulfotransferase activities, which differ in pH dependent profiles, pH optima (7.6 and 7.0, respectively) and the influence of Mn2+. PMID- 10815991 TI - Sialic acids in fungi: a minireview. AB - The increasing number of reports on the presence of sialic acids in fungi (N acetyl-, N-glycolyl- and 5,9-N,O-diacetylneuraminic acids) based on direct and indirect evidence warrants the present review. Formerly suggested as sialidase sensitive sources of anionic groups at the cell surface of fungal species grown in chemically defined media (e.g., Fonsecaea pedrosoi), sialic acids have also been found in Sporothrix schenckii, Paracoccidioides brasiliensis, Cryptococcus neoformans and recently, in Candida albicans. Methods used involved adequate hydrolysis and extraction procedures, HPTLC, gas-chromatography, colorimetry, mass spectroscopy, lectin and influenza virus binding. Apart from protecting fungal cells against phagocytosis (S. schenckii, C. neoformans) and playing a cellular structural role (F. pedrosoi), other biological functions of sialic acids are still being investigated. PMID- 10815992 TI - Development of monoclonal antibodies against bovine mucin core 2 beta6 N acetylglucosaminyltransferase. AB - Molecular cloning techniques have been used to produce abundant amounts of recombinant glycosyltransferases for biochemical studies. We recently cloned a cDNA which encoded bovine mucin core 2 beta6N-acetylglucosaminyl transferase (C2TF). Poly-histidine-C2TF fusion protein was generated from the cloned cDNA in the E. coli Xpress system and used to produce monoclonal antibodies (MAbs). We obtained seven hybridomas which secreted MAbs against bovine C2TF in mouse ascites with titers ranging from 1:1280 to 1:40960 as assessed by immunofluorescence assay (IF). Isotyping revealed that all seven MAbs were IgG (4 IgG1, 2 IgG2b and 1 IgG2a). The affinity constants (M(-1)) for these MAbs range from 5.4 x 10(7) to 1.2 x 10(9). These MAbs recognized bovine C2TF in tissue sections and on Western blottings. Six of these MAbs reacted with human core 2-M enzyme and one with both core 2-L and core 2-M enzymes on Western blottings. Therefore, these antibodies should be useful for further study of bovine and human core 2 enzymes. PMID- 10815993 TI - In vitro biological activities of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, coupled with N-acetylneuraminic acid. AB - In the previous study, N-acetylneuraminic acid (NANA) with C9 spacer was chemically coupled to human recombinant (rh) IL-1alpha in order to study the effect of glycosylation on its biological activities, and to develop IL-1 with less deleterious effects. In this study we examined a variety of IL-1 activities in vitro, including proliferative effect on T cells, antiproliferative effect on myeloid leukemic cells and melanoma cells, stimulatory effects on IL-6 synthesis by melanoma cells and PGE2 synthesis by fibroblast cells. NANA-introduced IL 1alpha (NANA-IL-1alpha) exhibited reduced activities about ten times compared with original IL-1alpha in all the activities performed in vitro. The competitive binding of 125I-IL-1alpha to mouse T cells and pre-B cells with unlabeled IL 1alphas suggests the decrease in binding affinities of NANA-IL-1alpha to both type I and type II IL-1 receptors. Therefore, reduced activities of NANA-IL 1alpha well correlated with the decrease in its receptor binding affinities. PMID- 10815994 TI - Coupling relations between molecular electronic and geometrical degrees of freedom in density functional theory and charge sensitivity analysis AB - The mapping concepts, reflecting the equilibrium, ground-state coupling between the electronic and geometrical degrees of freedom of both closed and open molecular systems are explored within density functional theory (DFT) and charge sensitivity analysis (CSA). After a brief overview of the principal derivatives and relations, including alternative Legendre transformed representations of molecular states, the explicit transformations are derived for the mapping between the nuclear coordinates Q, specifying the molecular geometry, and the 'electronic' coordinates represented by the electron density rho(r), in the local resolution, or the vector N = [N(alpha)] of the atomic electron populations, in the atoms-in-molecule (AIM) resolution. They fall into two categories: the electron following, transforming a given shift of the nuclear coordinates into the conjugate relaxation of the electronic 'coordinates' rho or N, and the electron preceding, 'translating' a specified displacement of these electronic degrees of freedom into the conjugate geometry relaxation. Algorithms for determining such transformations are discussed with a special emphasis placed upon the semiempirical CSA modeling in atomic resolution. Several additional mapping quantities are identified, which reflect the coupling between molecular electronic descriptors, e.g. the number of electrons or the chemical potential, and the relevant geometry-related quantities, e.g. Q or the forces acting on nuclei. Implications of the mapping concepts for chemistry are briefly examined and possible areas for their application are identified. PMID- 10815995 TI - Recognition of the electrophilic and nucleophilic centers in molecules via the radical charge transfer Fukui function AB - The asymptotic form of the radical charge-transfer Fukui function is analyzed as possible reactivity criterion. It is shown that atoms-in-molecules (AIM) discretisation of this function can be used to classify atoms of a given molecule into electrophilic and nucleophilic centers. Pyridine and 2-aminopyridine are taken as an illustrative example. The obtained numerical results are in accord with experimentally observed trends towards electrophilic/nucleophilic substitution. PMID- 10815996 TI - Density functional study of the S0 (X1Ag) and T1 (a3Au) states of the glyoxal molecule AB - The density functional theory (DFT) calculations in different approximations have been performed for the geometries and vibrational states of the trans-glyoxal molecule in the ground state S0 (X1Ag) and in the lowest excited triplet state T1 (a3Au, n-pi* type). Eight typical combinations of exchange and correlation functionals have been used. Comparative Hartree-Fock (HF) calculations have also been performed. For the open shell a3Au state the standard spin-unrestricted Hartree-Fock and Kohn-Sham approaches (UHF, UKS) as well as the restricted open shell versions (ROHF, ROKS) have been applied. The calculated frequencies have been compared, among others, with the data resulting from the most recent phosphorescence exicitation spectra of glyoxal cooled in the supersonic molecular beam, recorded with the cooperation of one of us (JH) for the spin-forbidden S0 T1 transition. The most realistic description of the vibrational frequencies, within the unscaled harmonic approximation, can be obtained using the 3-parameter Becke-93 exchange functional (B3), whereas this description practically does not depend on the correlation functional used. Our calculations support the recently reexamined experimental energy of the symmetric CH-rocking fundamental for the T1 state. PMID- 10815997 TI - Effective fragment potentials and the enzyme active site. AB - Optimization of the binding conformation of a substrate in an enzyme active site using ab initio quantum chemistry methods are intractable since the active site comprises several hundred atoms. However, the active site can be decomposed into an active and spectator region where the spectator residues are represented by effective fragment potentials and reducing the number of all-electron atoms involved in the chemistry to a reasonable level. The effective fragment potentials for electrostatics and polarization are implemented in GAMESS but the repulsive and charge transfer potentials are fit to interaction energies of water with models of the residues. These repulsive/charge transfer potentials are generated for the protein residues and the EFP are then used to optimize binding of a transition state analogue to chorismate mutase (B. subtilis) and small dianions to ribonuclease A. For chorismate mutase the calculated binding conformation compares well to the comparable X-ray structure. The binding of the inhibitor to the glutamate/glutamine mutant active site is then predicted with the optimization including the glutamine residue constrained only at the C alpha atom. The binding conformations suggest important roles for tyr108 and arg63, which have not been noted earlier. The electrostatic stabilization of the transition state by the active site charge distribution has to be augmented by a specific electronic activation by glu78. In ribonuclease A, the protons are found to move to provide a clustering of the charges to bind the small dianions, phosphate, thiophosphate, and sulfate. PMID- 10815998 TI - Chemical fragmentation in quantum mechanical methods. AB - We give a survey on the application of the chemical fragmentation concept in computer modelling of extended covalent systems. It will be stressed that information on molecular topology, as well as location and composition of the reaction centre allows the construction of a reasonable initial guess for the wave function and thus facilitates the solution of the Schrodinger equation. For systems, where the chemical changes are localised to a few atoms, while others play the role of essentially electrostatic perturbation, a partition into active site and environment is possible providing a background to hybrid quantum mechanical/molecular mechanical (QM/MM) methods. Full molecular orbital treatment of large covalent systems at the minimal basis, semiempirical level becomes possible in the frame of the fragment self-consistent field (FSCF) method which was developed in the past two decades in our laboratory. As an application, we discuss the hydride shift reaction step in xylose isomerase catalysis. PMID- 10815999 TI - Absolute and relative energies from polarized atomic orbital self-consistent field calculations and a second order correction. Convergence with size and composition of the secondary basis AB - Polarized atomic orbitals (PAO's) are molecule-adapted minimal basis functions that are variationally obtained as an atom-blocked transformation from a conventional extended basis set, as a Hartree-Fock calculation is performed in the PAO basis. This approximation yields a higher energy than a HF calculation performed in the extended basis, although the two results converge to the same limit as the extended basis approaches completeness on each atom. To test the rate of convergence, PAO-HF calculations were performed using cc-pVXZ and aug-cc pVXZ basis sets for the water monomer and dimer, and six substituted ethylenes. The results show that the quality of PAO calculations converges smoothly with X. The use of augmented functions is recommended. To correct a PAO-HF calculation for residual deficiencies, a noniterative second order correction is introduced. This correction corresponds to an energy-weighted steepest descent step, and substantially improves the quality of PAO energies. PMID- 10816000 TI - Solvent effect on hydrogen bonded ammonia-hydrogen halide complexes: continuum medium versus cluster models AB - The influence of water and dinitrogen environment on the structures of hydrogen bonded XH-NH3 (X = F, Cl, and Br) complexes has been investigated using the self consistent isodensity polarizable continuum solvent model (SCI-PCM) with two values of dielectric constants, 1.5 and 78.0, respectively. The geometrical parameters, along with the interaction energies as well as harmonic frequencies were calculated at the B3M-LYP/6-311 + + G(d,p) level of theory. The results from dielectric continuum model were compared with those obtained by traditional supermolecule approach. In order to model the proton transfer in XH-NH3 systems by supermolecular approach, one should add from two to four explicit water or dinitrogen molecules to the calculations. The dielectric continuum model provides similar effect using less effort. As our results shown, the SCI-PCM model, where a solvent is treated as a continuum medium, reproduces quite accurately the molecular properties of investigated medium strong hydrogen bonded systems both with dinitrogen and water environment. In order to represent solvent effects on hydrogen-bonded complexes with less acidic HX subunit it is necessary to take explicit solvent molecules into account. PMID- 10816001 TI - A theoretical study on reaction pathways to carbanions. AB - Different substituents (NO2, C6H5, NH2, NH-CH=CH-CHO) to a methylene group were taken into account to investigate under which conditions the mechanism of formation of carbanions by proton transfer to a base (methylamine) can be favorable, as a preliminary study of the reaction catalyzed by semicarbazide sensitive amine oxidases. Three different approaching paths of methylamine to C(alpha) in NO2-C(alpha)H2-NO2, and the relevant potential energy surfaces, were examined at the SCF/3-21G and 6-31G* levels. The proton transfer along the first two paths occurred with a similar barrier, which became fairly consistent after including the MP2 correlation correction, with either basis set, while the last approaching path was abandoned. For the other model systems the minimum was searched only at the 3-21G level in the vicinity of the first reaction path. The substitution of a nitro group with a phenyl group sharply raised the barrier for the proton transfer to methylamine. Also by substituting the second nitro group with either -NH2 or -NH-CH=CH-CHO, a steep uphill pathway was found. A more realistic model of the substrate-cofactor complex, namely the Schiff base between benzylamine and pyridoxal, again produced a barrier, almost matching that obtained for C6H5-C(alpha)H2-NO2. In both cases, the energy profiles for the rotation about the CC(alpha)NC dihedral and the proton shift tautomers were also considered at the 3-21G and 6-31G* levels. A preliminary scan of the effect of methyl (or methylphosphate) substitutions to the pyridoxal ring was performed and the stability of the Schiff bases involving other cofactors was also considered. PMID- 10816002 TI - Computer experiments on xenon-containing molecules AB - Quantum chemical calculations have been performed on xenon-containing rare gas molecules. The novel HXeY molecules are best characterised as HXe+Y- where Y can be at least H, Cl, Br, I, CN, SH, OH or NCO. A good agreement of MP2/LJ18/6-311 + + G(2d,2p) results is obtained with respect to the trend in experimental data for the Xe-H stretching wavenumbers of the HXeY molecules. Predictions concerning compounds between Xe and carboxylic acids are made. It is shown that Xe can bind to the carboxy group of the side chains of amino acids, thus, allowing in principle Xe to bind to proteins. PMID- 10816003 TI - Specificity of damage recognition and catalysis of DNA repair. AB - A common feature of DNA repair enzymes is their ability to recognize the damage independently of sequence in which they are found. The presence of a flipped out base inserted into the protein in several DNA-enzyme complexes suggests a contribution to enzyme specificity. Molecular simulations of damaged DNA indicate that the damage produces changes in DNA structure and changes the dynamics of DNA bending. The reduced bending force constant can be used by the enzyme to induce DNA bending and facilitate base flipping. We show that a thymine dimer (TD) containing DNA requires less energy to bend, lowering the barrier for base flipping. On the other hand, bending in DNA with U-G mismatch is affected only by a small amount and flipping is not enhanced significantly. T4 endonuclease V (endoV), which recognizes TD, utilizes the reduced barrier for flipping as a specific recognition element. In uracil DNA glycosylase (UDG), which recognizes U G mismatches, base flipping is not enhanced and recognition is encoded in a highly specific binding pocket for the flipped base. Simulations of UDG and endoV in complex with damaged DNA provide insight into the essential elements of the catalytic mechanism. Calculations of pKas of active site residues in endoV and endoV-DNA complex show that the pKa, of the N-terminus is reduced from 8.01 to 6.52 while that of Glu-23 increases from 1.52 to 7.82. Thus, the key catalytic residues are in their neutral form. The simulations also show that Glu-23 is also H-bonded to O4' of the 5'-TD enhancing the nucleophilic attack on Cl and that Arg 26 enhances the hydrolysis by electrostatic stabilization but does not participate in proton transfer. In the enzyme-substrate complex of UDG, the role of electrostatic stabilization is played by His-268, whose pKa increases to 7.1 from 4.9 in the free enzyme. The pKa of Asp-145, the other important catalytic residue, remains around 4.2 in the free enzyme and in the complex. Thus, it can not act as a proton acceptor. In the complex the 3'-phosphate of uracil is stabilized next to Asp-145 by two bridging water molecules. Such a configuration activates one water molecule to act as a proton acceptor to produce a stabilizing hydronium ion and the other as a proton donor to produce the nucleophilic hydroxide. It appears that DNA glycosylases share commonalties in recognition of damage but differ in their catalytic mechanisms. PMID- 10816004 TI - Interaction of Ia and IIa group cations with the guanine site in cytosine-guanine nucleic acid base pair: an ab initio Hartree Fock study in the absence of basis set superposition error. AB - Structures and energetics of complexes between guanine...cytosine Watson Crick (GCWC) DNA base pair and various metal cations were investigated by an ab initio Hartree Fock (HF) study in the absence of basis set superposition error. Cations were allowed to interact with N7 and O6 sites of guanine. The BSSE free gradient geometry optimisation were performed in the framework of the SCF-MI (self consistent field for molecular interactions) theory. In particular, the structure of the complex with the mono and bivalent cations, like H+, Na+, K+, Mg++, Ca++ were analysed showing that the coordination to the N7 and O6 sites of the GCWC pair can generate non-WC hydrogen bonding patterns. The results demonstrate that the a priori elimination of the BSSE allows to study molecular clusters of biological interest by employing small basis sets. PMID- 10816005 TI - Density functional MO calculation for stacked DNA base-pairs with backbones. AB - In order to elucidate the effect of the sugar and phosphate backbones on the stable structure and electronic properties of stacked DNA base-pairs, we performed ab initio molecular orbital (MO) calculations based on the density functional theory and Slater-type basis set. As a model cluster for stacked base pairs, we employed three isomers for the dimer unit of stacked guanine-cytosine pairs composed with backbones as well as base-pairs. These structures were fully optimized and their electronic properties were self-consistently investigated. By including the backbones, the difference in total energy among the isomers was largely enhanced, while the trend in relative stability was not changed. The effect of backbones on the electronic properties is remarkable: the MOs with the character of the PO4 parts of backbones appear just below the highest-occupied MO. This result indicates that the PO4 parts might play a rule as a reaction site in chemical processes concerning DNA. Therefore, we conclude that the DNA backbones are indispensable for investigating the stability and electronic properties of the stacked DNA base-pairs. PMID- 10816006 TI - New conjugated polyenes with 1,3-dialkyl-2-thiobarbituric acid moiety as materials for nonlinear optics: theoretical calculations, synthesis and spectral properties AB - In this work we report the results of our study on electronic and spectral properties of conjugated polyenes with electron-accepting 1,3-dialkyl-2 thiobarbituric acid moiety. In model calculations, we examine the effect of the conjugated polyene length on infrared (IR) and Raman spectra of the polyenes by means of ab initio HF/3-21G*. Nonlinear properties were also studied by AM1 method in frames of the sum-over-states (SOS) and finite-field formalism. It was concluded that in well-resolved IR and Raman spectra the frequencies and band intensities can provide valuable information relating to C=C bond lengths in polyene chain and relative polarizabilities. Near-linear correlation between polarizability and integral IR band intensity, corresponding to all C=C stretching modes, and the rather nonlinear relationship of polarizability with integral Raman activity, was found. In our calculation we predict that polarizability and the first hyperpolarizability increases with elongation of polyene chain while the second hyperpolarizability increases smoothly in a quadratic way. In contrast to the linear relationship between polarizability and polyene chain length the dipole moment versus chain length is predicted to be nonlinear. A good agreement was found between experimental and calculated Raman spectra of one newly synthesized compound studied. PMID- 10816007 TI - Electronic structure and nonlinear optical properties of model push-pull polyenes with modified indanone groups: a theoretical investigation AB - Several model polyenes with modified indanone groups were studied by means of density functional theory (DFT) B3LYP/6-31G*, ab initio HF/3-21G* and semiempirical AM1 methods. We investigated the effect of several substituents upon the relationship between the structure, spatial distribution of the highest occupied and the lowest unoccupied pi-MOs, a concept of the global softness and the global hardness as well as both linear and nonlinear polarizabilities for the set of pi-electron chromophores represented by the short-chain model polyene (butadiene) carrying out p-methoxyphenyl group on the one end and several modified indanone groups on the opposite end of the molecule. As probing endocyclic groups used to modify the structure of indanone the following substituents: > CH2; > C=O; > SO2, > C=CH(NO2) and > C=C(CN)2 were selected. The cubic relationship between the polarizability and the global softness was found. The highest polarizabilities (alpha, beta, gamma) are predicted for the derivatives with > C=C(CN)2 group. It was found that the value of beta depends mainly on the difference between dipole moments in the excited and ground states of the molecules. In the case of > SO2 group the results of AMI calculations significantly deviate from relationships found for other derivatives. Experimental IR and Raman spectra of newly synthesized indandione derivative of cinnamaldehyde were compared with computed ones. PMID- 10816008 TI - Molecular modeling of the catalytic domain of serine/threonine phosphatase-1 with the Zn2+ and Mn2+ di-nuclear ion centers in the active site. AB - Catalytic domains of the metalloenzymes protein phosphatases (PPP) 1, 2A and 2B (PP1, PP2A and PP2B, respectively) are homologous to approximately 45%, with the residues in the enzymatic centers strictly conserved. PP1, PP2A and PP2B are abundant in cells and they dephosphorylate serine and/or threonine residues in a variety of proteins serving as cellular phospho switches. The active enzymes work as invariant catalytic subunits PP1c, PP2Ac and PP2Bc, respectively, complexed with diverse regulatory subunits, dependent on the enzymes' specific location and biological function. The crystal structures of PP1c and PP2B (calcineurin) heterotetramer calcineurinA x calcineurinB x FKBP x FK506 have been determined. A comparison of the catalytic subunits of both enzymes indicates their significant structural homology and virtual identity within the catalytic centers, each including a set of conservative amino acids, two metal ions and a phosphate; thus confirming a hypothesis on their common enzymatic mechanisms. The elongated substrate cleft at the active centre is kinked by approximately 120 degrees at the active center in its middle and thus divided into a pre-phospho-Ser/Thr (ligand N-terminal) and a post-phospho-Ser/Thr (ligand C-terminal) section. In PP1c the N-terminal section is highly acidic while in PP2Bc is not. This feature is likely pertinent but not sufficient to the enzymes' selectivity, which is also controlled by regulatory subunits, diverse in various tissues. The metalloenzymes in general and PPP in particular are hard to deal with using theoretical simulations due to parameterization problems for the metal cations. In fact, there are only a few PP1c simulations reported, with the metal di-cations treated quite crudely. This is a preliminary work, in which we introduce and test against some experimental evidence a concept of pseudomolecules of proper geometry, composed of double metal (2Zn2+ or 2Mn2+) cation, and the OH- nuclephile incorporated into the PP1c catalytic site. Both models are associated with either the phosphate (a free enzyme) or the phosphorylated dodecapeptide RRRRPpTPAMLFR, an active fragment (residues 29-40) of a regulatory subunit DARPP-32 inhibitor (PP1c-inhibitor complex); four models total. We have parameterized both pseudomolecules within the AMBER force field. Subsequently, using molecular dynamic in water, we have found the free PP1c subunits to be less stable than the complexed ones and we have speculated on possible reasons for this feature. PMID- 10816009 TI - Ab initio calculations on the five-membered alumino-silicate framework rings model: implications for dissolution in alkaline solutions AB - An ab initio restricted Hartree-Fock calculation utilising the standard 6-31G basis set was used to calculate total energies after PM3 calculations of energy optimised geometries for the five-membered alumino-silicate framework rings cluster for a total of ten T sites. Calculations have shown that in the absence of protons or other ions, the most favourable sites for 1A1, 2A1 and 3A1 substitution of Si are the T6, T1 and T9 sites respectively. With more Al atoms replacing Si in a cluster, T-O bond lengths and T-O-T angles show lengthening and sharpening trends respectively, which indicates that the structure is distorted to a more relaxed symmetry with Obr atoms moving outwards. The calculated bond lengths and angles have been shown to match the values observed in previous studies, including those for a four-membered alumino-silicate single ring cluster. Based on the optimised five-membered alumino-silicate framework rings model, a further ab initio HF calculation has been conducted on ring breakage for releasing Al(Q3) and Si(Q3) centres to form T(OH)4 and HOT(OM)3 tetrahedra under local and highly alkaline environment. The obtained results suggest that Al(Q3) compared with Si(Q3) breaks more readily with the exothermal reaction enthalpy being in excess of -244.4 kJ/mol, while the most reactive Si(Q3) centre shows an exothermal reaction enthalpy of only -33.8 kJ/mol. This indicates that Al dissolves in preference to Si in local environment. The dissolution mechanism of the five-membered Al-Si framework rings model in highly alkaline solutions has been suggested to be composed of an ion-pairing reaction and an interaction between the remaining broken ring cluster triple bond TOH and MOH. PMID- 10816010 TI - Quantum chemical modeling (DFT) of active species on the V-W-O catalyst surface in various redox conditions AB - This study concerns quantum chemical modeling of water behaviour on V-W-O systems. It was undertaken in order to validate the hypothesis that the presence of W atoms on (001) surface of the crystalline vanadia-like species, facilitates low temperature water dissociation. Quantum chemical calculations were done with the use of modern electronic structure methodology, based on the density functional theory (DFT). The program package DMol of Molecular Simulations, was applied for the calculations. The calculations were performed for small clusters representing two adjacent metal sites, in pentacoordinated oxygen environment, analogous to bipiramidal clusters, introduced in description of the (001) layers of vanadium pentoxide. PMID- 10816011 TI - Characteristics of the ligand-binding site interaction for a series of arecoline derived muscarinic agonists: a quantum chemical study. AB - This work focuses on electronic and conformational structure of bicyclic analogues of arecoline and sulfoarecoline--muscarinic receptor agonists structurally related to 4,5,6,7-tetrahydroisoxazolo[4,5-c]pyridin-3-ol (THPO) and its S-methylsulfonium derivative (DHTO). Conformational freedom of six-member rings containing sulphur and nitrogen has been investigated by means of semiempirical AM1 method. Interaction between 'cationic heads' of two representative compounds and HCO2- ion serving as a model of carboxyl group of Asp in the muscarinic receptor has been modelled using DFT method in local approximation (LDA with VWN exchange correlation functional). Electrostatic potential (ESP) around studied complexes and ligands with added electron (simulation of complex formation) are presented and analysed. Position and depth of ESP minima in a series of studied ligands correlate well with their activity as muscarinic agonists. On the basis of our results the mechanism of ligand binding site interaction may be elucidated. The calculations allow also for the comparison of bicyclic analogues of arecoline with already existing model for muscarinic pharmacophore and for rationalization of model parameters. PMID- 10816012 TI - Improved QSARs for predictive toxicology of halogenated hydrocarbons. AB - In our continuing efforts to provide a predictive toxicology capability, we seek to improve QSARs (quantitative structure-activity relationships) for chemicals of interest. Currently, although semi-empirical molecular orbital methods are hardly the state of the art for studying small molecules, AM1 calculations appear to be the method of choice when calculating quantum-chemical descriptors. However, with the advent of modern computational capabilities and the development of fast algorithms, ab initio molecular orbital and first principles density functional methods can be expeditiously applied in current QSAR studies. We present a study on halogenated alkanes to assess whether more accurate quantum methods result in QSARs that correlate better with experimental data. Furthermore, improved QSARs can also be obtained through development of new descriptors with explicit physical interpretations that should lead to better understanding of the mechanisms involved in the toxic response. We show that descriptors calculated from chemical intermediates may be useful in future QSARs. PMID- 10816013 TI - The conformational characteristics of Congo red, Evans blue and Trypan blue. AB - The structures of the closely related bis-azo dyes Evans blue, Trypan blue and Congo red, which appeared to have different self-assembly properties and correspondingly different abilities to form complexes with amyloids and some other proteins, were compared in this work. Ab initio and semi-empirical methods were used to find the optimal structures and partial charge distributions of the dyes. The optimal structures were searched using different widely used programs. The structures of Congo red and evans blue were found to be planar, except for the torsion on the central diphenyl bond connecting the two halves of the dye. Both symmetrical parts of the molecules appeared very close to planarity. However, Trypan blue exhibits non planarity on the di-azo bonds, as well as on the central bond between the symmetrical parts of the dye. In a consequence, the non planarity of this molecule is higher than in the case of its isomer, Evans blue and Congo red as well. The extra rotation around the azo bonds extorted by the close proximity of the sulfonic groups may be the direct cause of its poor self-assembling and complexation properties versus Evans blue. PMID- 10816015 TI - Structural and dynamical properties of concentrated aqueous NaOH solutions: a computer simulation study AB - Molecular dynamics simulations of NaOH aqueous solutions have been performed with a flexible model of the water molecule. Ion-ion interactions have been described by the (12-6) Lennard-Jones type and coulomb potentials. The concentration of the solutions ranged from 0.5 to 19 M. The structures of hydration shells of the ions were analysed using the radial distribution functions, Voronoi tessellations, the running and O'Keeffe coordination numbers and other tools. Structural properties obtained from simulations agreed reasonably well with results of the available experimental data. The dynamical properties were calculated and compared with experimental measurements. A particularly careful analysis was applied to the topological properties of the ionic structures in solution. The results of calculations suggest an existence of the ordered structures of ions in solution and supports the Ruff theory of ionic quasi-lattices in concentrated solutions. PMID- 10816014 TI - Spatial distribution functions as a tool in the analysis of ribonucleic acids hydration--molecular dynamics studies. AB - The spatial distribution functions (SDFs) determined as three-dimensional density distribution of hydrogen and oxygen atoms of water in a local coordinate system linked with RNA molecule are used to study details of the spatial structure of aqueous solution around selected parts of RNA duplexes: r(CGCGCG)2 and 2'-O Me(CGCGCG)2. The influence of the 2'-O-methylation on the hydration pattern of RNA helical fragments is visualized at the atomic level. PMID- 10816016 TI - Potential traps for an excess electron in liquid water. Geometry, energy distributions and lifetime AB - The possibilities of trapping of an excess electron by potential fluctuations in liquid water have been investigated by means of the computer simulation method. The equilibrium configurations of 200 water molecules were generated by the molecular dynamics method. Given an interaction potential between a negative test charge and a water molecule, the molecular configurations generated by the simulation are searched for local minima of the potential energy. The analysis of a large set of the minima allows us to obtain an extensive statistical description of the microscopic trapping sites, including the distributions of the trap energy, volume and the trapping cross section. The estimated concentration of the electron traps in liquid water is about 0.5 mol/dm3. The possiblity of electron trapping depends very strongly on the lifetime of the potential traps. The simulations yielded the distribution of the trap lifetime with the average of 84 fs. A substantial fraction (20%) of the traps live longer than 100 fs, a small fraction (0.2%) live as long as 1 ps. These values can be compared with experimental measurements of the electron hydration time of the order of 100 fs. PMID- 10816017 TI - Monte Carlo simulations of I2- (CO2)16 and I2- (N2O)16 clusters. Minimum energy structures and solvation energy AB - Structures of the I2- (CO2)16 and I2- (N2O)16 clusters have been studied along the dissociation path of the halide ion. Minimum energy structures and solvation energies have been estimated. The influence of the electrostatic interaction on the I2- charge distribution and the structure of the solvation shell has been discussed. It has been shown that the small dipole moment of the N2O molecule does not have much influence on the cluster properties, and the large quadrupole moments of both solvent clusters give the major contribution into electrostatic interactions. The structure of the clusters is more sensitive to different potential models than to the dipole moment of the N2O molecule. PMID- 10816018 TI - A global optimization strategy for predicting alpha-helical protein tertiary structure. AB - We present a global optimization strategy that incorporates predicted restraints in both a local optimization context and as directives for global optimization approaches, to predict protein tertiary structure for alpha-helical proteins. Specifically, neural networks are used to predict the secondary structure of a protein, restraints are defined as manifestations of the network with a predicted secondary structure and the secondary structure is formed using local minimizations on a protein energy surface, in the presence of the restraints. Those residues predicted to be coil, by the network, define a conformational sub space that is subject to optimization using a global approach known as stochastic perturbation that has been found to be effective for Lennard-Jones clusters and homo-polypeptides. Our energy surface is an all-atom 'gas phase' molecular mechanics force field, that is combined with a new solvation energy function that penalizes hydrophobic group exposure. This energy function gives the crystal structure of four different alpha-helical proteins as the lowest energy structure relative to other conformations, with correct secondary structure but incorrect tertiary structure. We demonstrate this global optimization strategy by determining the tertiary structure of the A-chain of the alpha-helical protein, uteroglobin and of a four-helix bundle, DNA binding protein. PMID- 10816019 TI - From fold recognition to homology modeling: an analysis of protein modeling challenges at different levels of prediction complexity. AB - An analysis of different approaches to protein structure prediction is presented based solely on the range of models submitted to the third Critical Assessment of Protein Structure Prediction (CASP3) conference. CASP conferences evaluate the current state of the art of protein structure prediction by comparing blind prediction efforts of many groups for the same set of target sequences. Target sequences may be highly similar to those with known structure or can be totally (at least superficially) sequentially dissimilar. Techniques applied to those blind predictions (over 40 targets) ranges from a detailed homology prediction to the detection of remote homologues well below a twilight zone of protein sequence similarity. For the CASP3 conference, we have submitted predictions, totaling 35, with various levels of difficulty and complexity. For ten submitted homology targets, eight of them were determined by experiment so far. The RMSD of C-alpha atoms are 1.2-1.7, 2.3, and 4.6-17.9 A for the three easy targets, two hard targets, and three very hard homology targets, respectively. Out of 18-fold recognition predictions available for analysis, we got six correct predictions, five near misses, three tough near misses and four far misses. Here we analyze successes and failures of those predictions in an attempt to identify common problems and common achievements. PMID- 10816020 TI - From fold to function predictions: an apoptosis regulator protein BID. AB - With the rapidly increasing pace of genome sequencing projects and the resulting flood of predicted amino acid sequences of uncharacterized proteins, protein sequence analysis, and in particular, protein structure prediction is quickly gaining in importance. Prediction algorithms can be used for preliminary annotation of newly sequenced proteins and, at least in some cases, provide insights into their function and specific mode of action. Such annotations for several microbial genomes were performed by several groups and placed in public domain for evaluation. An example presented in this work comes from a related project of structural and functional predictions for proteins involved in the process of controlled cell death (apoptosis). The BID protein belongs to an important class of regulators of apoptosis identified by short sequence motifs. Here, several fold prediction methods are used to build a series of three dimensional models. Structure analysis of the models with reference to the biological data available allows selection of the most appropriate model. It is found that the most likely structural model of BID is built on the structure of Bcl-X(L). The model is discussed in terms of experimental data on specific proteolytic cleavage of BID and its effect on BID interactions with other proteins and membranes. PMID- 10816021 TI - LFP: A PC-program for ligand-field analysis of 3d(n) ions in Oh and lower symmetries AB - A modular and efficient version of PC program for calculating ligand-field parameters, i.e. crystal-field model (CFM) and angular overlap model (AOM) is presented. The LFP program is designed to calculate the ligand-field parameters of low symmetry transition metal complexes. It is based on the general method for the analysis of central ion states distortion using group theory. The program has not closed form. It will be extended, according to spectroscopic studies in our laboratory. It is written in FORTRAN language. PMID- 10816022 TI - SYMTERM--program for modelling chemical processes in non-isothermal conditions AB - The program SYMTERM allows to perform the numerical simulations of chemical processes which take place in the thermal analysis apparatus, considering two important factors: the exchange of heat between a sample and an environment, and the influence of heat evolved in the reaction on its kinetics. The program makes it possible to obtain kinetics profiles (DTA, TG, DTG or DSC curves) at various heating rates and permits to determine both the activation energy Eact and the reaction order of a single rate-limited step reaction with using DTA (DSC) and TG data simultaneously. PMID- 10816023 TI - Nonlinear regression in parameter estimation from polarographic signals AB - In this work we describe a detailed treatment of polarographic data curves, including error analysis, by means of nonlinear least-squares in its standard form (or resorting to the errors in variables model). Error estimates for the related parameters are additionally verified by Monte-Carlo simulation and resampling techniques. PMID- 10816024 TI - Mechanisms of osteoporosis after hematopoietic cell transplantation. AB - Osteopenia and osteoporosis are common complications of bone marrow and peripheral blood stem cell transplantation. Bone loss occurs in 50% to 60% of patients treated with the most common preparatory regimens. The major causes of transplant-related bone loss are primary hypogonadism (low estrogen and testosterone), secondary hyperparathyroidism due to low serum calcium, and posttransplant steroid therapy. Other transplant-related treatments that induce bone loss are discussed. Trabecular bone is particularly vulnerable to transplant related therapies. The spine and hip contain 50% to 75% trabecular bone and are most at risk for fracture after hematopoietic cell transplantation (HCT). The structure of bone and the bone cells that are involved in maintaining skeletal integrity are discussed, followed by a discussion of the transplant-related therapies that have been shown to cause damage to bone and lead to bone loss. Recommendations for patients undergoing HCT include (1) evaluation of bone mineral density either shortly before or shortly after transplantation and appropriate intervention and monitoring based on the results; and (2) evaluation of estrogen and testosterone levels after HCT and replacement when appropriate; and (3) administration of bisphosphonate therapy to all patients on steroids for >2 months. Early intervention and prevention of bone loss can have a tremendous clinical impact for patients undergoing HCT because once significant bone loss has occurred, it is difficult to reverse. PMID- 10816025 TI - Osteoporosis after blood and marrow transplantation: clinical aspects. PMID- 10816026 TI - An immunoablative regimen of fludarabine and cyclophosphamide prevents fully MHC mismatched murine marrow graft rejection independent of GVHD. AB - The prevention of graft rejection in the setting of nonmyeloablative transplant approaches might be mediated by chemotherapy-induced host immunoablation and by the graft-promoting effects of graft-versus-host disease (GVHD). To evaluate whether host immunoablation alone might allow for alloengraftment, we developed an F1-into-parent murine marrow rejection model using host preparative regimens of lethal total body irradiation (TBI; 950 cGy), sublethal irradiation (600 cGy), or combinations of fludarabine (Flu) and cyclophosphamide (Cy). A preparative regimen selectivity index (SI) was calculated to determine whether host lymphocytes were preferentially depleted relative to myeloid cells (SI = number of host myeloid/number host T lymphoid cells remaining after preparative regimen administration). Saline-treated recipients were assigned an SI value of 1.0. Recipients of lethal TBI had reduced myeloid cells relative to T cells (SI = 0.6). In contrast, all Flu/Cy regimens preferentially depleted host T cells: recipients of Flu (100 mg/kg per day)/Cy (50 mg/kg per day) for 10 days (SI = 28.1); recipients of Flu (100 mg/kg per day)/Cy (100 mg/kg per day) for 10 days (SI = 64.1); and recipients of Flu (100 mg/kg per day)/Cy (50 mg/kg per day) for 19 or 27 days (SI = 74.6). The 10-day regimen of Flu/Cy (50 mg/kg per day) did not severely reduce host T cell numbers, nor did it prevent F1 marrow rejection (<1% chimerism, n = 14). In contrast, the 10-day regimen of Flu/Cy (100 mg/kg per day) reduced T-cell numbers below that of lethal TBI recipients and prevented F1 marrow rejection (11.4% chimerism, n = 15); donor chimerism was predominant in lymphoid cells and was stable through day 240 post-BMT. Additionally, the 19- or 27-day regimen of Flu/Cy, which most selectively depleted host T cells, also prevented F1 marrow rejection (6.3% chimerism, n = 15). These results therefore demonstrate that optimized Flu-containing, immunoablative preparative regimens can prevent fully MHC-disparate marrow rejection independent of GVHD. PMID- 10816027 TI - Tacrolimus does not abrogate the increased risk of acute graft-versus-host disease after unrelated-donor marrow transplantation with allelic mismatching at HLA-DRB1 and HLA-DQB1. AB - One hundred patients of median age 34 years (range, 14-53) received bone marrow transplants from unrelated donors serologically matched for human leukocyte antigen HLA-A, HLA-B, and HLA-DR using tacrolimus and minimethotrexate for prevention of acute graft-versus-host disease (GVHD). Sixty-eight patient-donor pairs had allelic matches at HLA-DRB1 and HLA-DQB1, 20 pairs had a single mismatch at HLA-DRB1 or HLA-DQB1, and 12 were mismatched at both HLA-DRB1 and HLA DQB1. Minimum follow-up time was 6 months. Grades 2 to 4 GVHD occurred in 43% of patients with matched donors, 69% with single allele-mismatched donors, and 71% with double allele-mismatched donors; grades 3 to 4 GVHD occurred in 22%, 43%, and 64%, respectively. On multivariate analysis, the relative risk of grades 2 to 4 GVHD was 2.2 (95% CI, 1.1-4.5; P = .03) with a single allele mismatch and 2.7 (95% CI, 1.2-6.0; P = .02) with a double allele mismatch. The relative risks of grades 3 to 4 GVHD were 3.0 (95% CI, 1.2-7.6; P = .02) and 5.0 (95% CI, 1.9-12.6; P = .001), respectively. Day 100 treatment-related mortality was also adversely affected by allelic mismatching, occurring in 21% of those with matched donors, 50% with single allele-mismatched donors, and 42% with double allele-mismatched donors (P = .02), but overall survival at day 180 did not differ significantly among the 3 groups. Tacrolimus does not abrogate the adverse impact of allele mismatching at HLA-DRB1 and HLA-DQB1 on the risk of moderate-to-severe acute GVHD. PMID- 10816028 TI - Cardiac toxicity following high-dose cyclophosphamide, cisplatin, and BCNU (STAMP I) for breast cancer. AB - We retrospectively evaluated 443 breast cancer patients treated with high-dose cyclophosphamide, cisplatin, and BCNU (STAMP-I) with autologous stem cell support to characterize the cardiac toxicity of this regimen. Patients had stage II-III (n = 243) or stage IV (n = 200) breast cancer. We observed an overall 5.1% incidence of cardiac complications, both clinical and subclinical, in the whole group: 4.9% in stage II-III and 5.5% in stage IV patients. Clinical cardiomyopathy (CMP) was observed in 1.6% of stage II-III patients (1 case of fatal grade 5 toxicity and 3 cases of grade 3 CMP) and in 3.5% of patients with stage IV disease (1 case of grade 4 and 6 cases of grade 3). The incidence of cardiac toxicity did not differ significantly between the groups. Prior radiation therapy to the mediastinum or left chest wall (P = .001) and advanced age (P = .01) were independent predictors of an increased risk of the appearance of this complication. No pharmacodynamic correlation was observed between any of the 3 drugs and cardiac toxicity. PMID- 10816029 TI - Autologous blood and marrow transplantation in patients 60 years and older. AB - Although many hematologic malignancies are more common in older patients, autologous blood and marrow transplantation (ABMT) has traditionally been restricted to patients younger than 60 years because of concerns that older patients would be either unable to provide a graft or unable to tolerate the therapy. From June 1995 to May 1998, 30 patients > or = 60 years underwent ABMT at our institution for low-grade lymphoma (4 patients), relapsed intermediate grade lymphoma (17 patients), or multiple myeloma (9 patients). The median patient age was 62.5 years (range 60-73). Pretransplantation conditioning regimens were CBV (cyclophosphamide, BCNU [carmustine], etoposide) or BEAM (carmustine, etoposide, cytarabine, melphalan) for intermediate-grade lymphoma patients and melphalan 140 mg/m2 + etoposide 60 mg/kg + total body irradiation 500 cGy for the others. The rescue product was bone marrow (BM; 4 patients), peripheral blood stem cells (PBSC; 23 patients), or BM+PBSC (3 patients). The median number of CD34+ cells/kg infused was 3.60 x 10(6) (range 0.53-31.0), by the International Society for Hematotherapy and Graft Engineering method. The treatment-related mortality at day 100 and at 6 months was 10% and 16.7%, respectively. The median days to neutrophil > 0.5 x 10(9)/L was 11 (range 9-25) and platelets > 20 x 10(9)/L was 16 (range 6-70). Three patients died of infection (days 26, 27, and 38), and 1 died of an intracranial hemorrhage related to persistent thrombocytopenia (day 130). Bearman regimen-related toxicity was moderate, with most toxicities < or = grade 2. Seven patients developed significant gut toxicity: 4 patients with Clostridium difficile colitis and 3 patients with neutropenic enterocolitis. Depressive symptoms and signs were noted in 4 patients. Three male patients developed decreased gonadal function after transplantation. These transplantations accounted for 997 patient days, of which 266 days (27%) were in the outpatient BMT program--a smaller percentage than in patients < 60 years (56%, P = .002). Twenty patients are alive 153 to > or = 1224 days after transplantation. ABMT in patients > or = 60 years of age is feasible. Further studies addressing supportive care particular to older patients and comparisons of ABMT with traditional approaches to multiple myeloma and relapsed non-Hodgkin's lymphoma in older patients are needed. Further work to identify elderly patients most likely to benefit from this approach is also required. PMID- 10816030 TI - Role of splenic irradiation in patients with chronic myeloid leukemia undergoing allogeneic bone marrow transplantation. Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation. PMID- 10816031 TI - Evidence-based assessment of diagnostic tests for ventilator-associated pneumonia. Executive summary. PMID- 10816032 TI - Panel methodology: Analytical principles in evaluating the performance characteristics of diagnostic tests for ventilator-associated pneumonia. PMID- 10816033 TI - Epidemiology of ventilator-associated pneumonia. AB - In summary, the method of diagnosis used for VAP accounts for reported differences in etiology, pathogenesis, and outcomes. Further studies are needed to assess outcomes related to various diagnostic methods rather than to assess the sensitivity and specificity of these methods. PMID- 10816034 TI - Radiologic diagnosis of ventilator-associated pneumonia. PMID- 10816035 TI - Clinical criteria in the diagnosis of ventilator-associated pneumonia. PMID- 10816036 TI - Endotracheal aspiration in the diagnosis of ventilator-associated pneumonia. PMID- 10816037 TI - Bronchoscopic BAL in the diagnosis of ventilator-associated pneumonia. PMID- 10816038 TI - Protected-specimen brush technique in the diagnosis of ventilator-associated pneumonia. PMID- 10816039 TI - Blinded invasive diagnostic procedures in ventilator-associated pneumonia. PMID- 10816040 TI - Bronchoscopy in nonresolving nosocomial pneumonia. PMID- 10816041 TI - Sulfide:quinone oxidoreductase in membranes of the hyperthermophilic bacterium Aquifex aeolicus (VF5). AB - The sulfide-dependent reduction of exogenous ubiquinone by membranes of the hyperthermophilic chemotrophic bacterium Aquifex aeolicus (VF5), the sulfide dependent consumption of oxygen and the reduction of cytochromes by sulfide in membranes were studied. Sulfide reduced decyl-ubiquinone with a maximal rate of up to 3.5 micromol (mg protein)(-1) min(-1) at 20 degrees C. Rates of 220 nmol (mg protein)(-1) min(-1)] for the sulfide-dependent consumption of oxygen and 480 nmol (mg protein)(-1) min(-1) for the oxidation of sulfide at 20 C were estimated. The reactions were sensitive towards 2-n-nonyl-4-hydroxyquinoline-N oxide, but insensitive towards cyanide. Both reduction of decyl-ubiquinone and consumption of oxygen by sulfide rapidly increased with increasing temperature. For the sulfide-dependent respiratory activity, a sulfide-to-oxygen ratio of 2.3+/-0.2 was measured. This indicates that sulfide was oxidized to the level of zero-valent sulfur. Reduction of cytochromes by sulfide was monitored with an LED array spectrophotometer. Reduction of cytochrome b was stimulated by 2-n-nonyl-4 hydroxyquinoline-N-oxide in the presence of excess sulfide under oxic conditions. This "oxidant-induced reduction" of cytochrome b suggests that electron transport from sulfide to oxygen in A. aeolicus employs the cytochrome bc complex via the quinone pool. Comparison of the amino acid sequence with the sequence of the sulfide:quinone oxidoreductase from Rhodobacter capsulatus and of the flavocytochrome c from Allochromatium vinosum revealed that the sulfide:quinone oxidoreductase from A. aeolicus belongs to the glutathione reductase family of flavoproteins. PMID- 10816042 TI - Cell wall and cytoskeleton reorganization as the response to hyperosmotic shock in Saccharomyces cerevisiae. AB - Transfer of exponentially growing cells of the yeast Saccharomyces cerevisiae to hyperosmotic growth medium containing 0.7-1 M KCl, 1 M mannitol, and/or 1 M glycerol caused cessation of yeast growth for about 2 h; thereafter, growth resumed at almost the original rate. During this time, formation of fluorescent patches on the inner surface of cell walls stained with Primulin or Calcofluor white was observed. The fluorescent patches also formed in solutions of KCl or when synthesis of the cell wall was blocked with cycloheximide and/or 2 deoxyglucose. The patches gradually disappeared as the cells resumed growth, and the new buds had smooth cell walls. Electron microscopy of freeze-etched replicas of osmotically stressed cells revealed deep plasma membrane invaginations filled from the periplasmic side with an amorphous cell wall material that appeared to correspond to the fluorescent patches on the cell surface. The rate of incorporation of D-[U-14C]glucose from the growth medium into the individual cell wall polysaccharides during osmotic shock followed the growth kinetics. No differences in cell wall composition between osmotically stressed yeast and control cells were found. Hyperosmotic shock caused changes in cytoskeletal elements, as demonstrated by the disappearance of microtubules and actin microfilaments. After 2-3 h in hyperosmotic medium, both microtubules and microfilaments regenerated to their original polarized forms and the actin patches resumed their positions at the apices of growing buds. The response of S. cerevisiae strains with mutations in the osmosensing pathway genes hog1 and pbs2 to hyperosmotic shock was similar to that of the wild-type strain. We conclude that, besides causing a temporary disassembling of the cytoskeleton, hyperosmotic shock induces a change in the organization of the cell wall, apparently resulting from the displacement of periplasmic and cell wall matrix material into invaginations of the plasma membrane created by the plasmolysis. PMID- 10816043 TI - Morphology-related effects on gene expression and protein accumulation of the yeast Arxula adeninivorans LS3. AB - The dimorphism of the yeast Arxula adeninivorans LS3 is regulated by cultivation temperatures. Up to 42 degrees C the yeast grows as budding cells, which turn to mycelia at higher temperatures. To test whether the dimorphism is exclusively induced by high temperatures or also by other conditions, mutants were selected with an altered behaviour with respect to dimorphism. After mutagenesis with N methyl-N'-nitro-N-nitrosoguanidine, five of 25,000 colonies formed a very rough surface consisting of mycelia at 30 degrees C, in contrast to the wild-type. These mutants allow temperature-mediated and morphology-related effects on gene expression and protein accumulation to be distinguished. Budding cells and mycelia showed different expression of genes encoding secretory proteins at the same temperature. Mycelia secreted two-fold more protein than budding cells, including the enzymes glucoamylase and invertase. This indicated that morphology, rather than temperature, is the decisive factor in the analysed processes. PMID- 10816044 TI - Modification of plasma membrane lipid order and H+-ATPase activity as part of the response of Saccharomyces cerevisiae to cultivation under mild and high copper stress. AB - Plasma membrane lipid disorganization takes place in cells of Saccharomyces cerevisiae grown under copper stress, as shown by fluorescence anisotropy measurements with the lipid reporter probe 1,6-diphenyl-1,3,5-hexatriene. The extent of plasma membrane disorganization, presumably due to copper-induced lipid peroxidation, was discontinuous when measured in cells grown in media supplemented with different concentrations of CuSO4. Results suggested the existence of adaptive mechanisms that cells employ to protect themselves against the deleterious effects of copper. The adaptive mechanisms examined in this study included the coordinate increase in the activities of Cu,Zn-superoxide dismutase (up to five-fold), glutathione reductase (up to 1.7-fold), and plasma membrane H+ ATPase (up to three-fold). These enzyme activities showed maximal levels in cells grown with copper supplied at intermediate concentrations, within the range that allowed growth. Significantly, at these concentrations, plasma membrane disorganization did not increase when increasing amounts of CuSO4 were supplied. However, at copper concentrations close to the maximal that allowed growth, the capacity of the yeast cell response to cope with the deleterious effects of copper was exceeded; plasma membrane lipid organization and plasma-membrane-bound H+-ATPase activity drastically declined in response to the increased levels of copper stress and the consequences on growth kinetics were even more severe. Our results clearly suggest that modification of plasma membrane H+-ATPase activity is either part of or the result of the global response of yeast to mild or high copper stress. PMID- 10816045 TI - Rhodoferax antarcticus sp. nov., a moderately psychrophilic purple nonsulfur bacterium isolated from an Antarctic microbial mat. AB - A new species of purple nonsulfur bacteria isolated from an Antarctic microbial mat is described. The organism, designated strain ANT.BR, was mildly psychrophilic, growing optimally at 15-18 degrees C with a growth temperature range of 0-25 degrees C. Cells of strain ANT.BR were highly motile curved rods and spirals, contained bacteriochlorophyll a, and showed a multicomponent in vivo absorption spectrum. A specific phylogenetic relationship was observed between strain ANT.BR and the purple bacterium Rhodoferax fermentans FR2T, and the two organisms shared several physiological and other phenotypic properties, with the notable exception of growth temperature optimum. Tests of genomic DNA hybridization, however, showed Rfx. fermentans FR2T and strain ANT.BR to be genetically distinct bacteria. Because of its unique set of properties, especially its requirement for low growth temperatures, we propose to recognize strain ANT.BR as a new species of the genus Rhodoferax, Rhodoferax antarcticus, named for its known habitat, the Antarctic. PMID- 10816046 TI - Identification and localization of the carboxysome peptide Csos3 and its corresponding gene in Thiobacillus neapolitanus. AB - Four genes encoding carboxysome shell peptides (csoS1A, csoS1B, csoS1C, csoS2), the genes encoding the large and small subunits of RuBisCO (cbbL, cbbS), and three unidentified ORFs constitute an operon in Thiobacillus neapolitanus. An unidentified ORF 1.54 kb in size is predicted from sequence analysis to encode a protein with a molecular mass of approximately 57 kDa. When this ORF was expressed in Escherichia coli under the control of its endogenous ribosome binding site, no peptide product was observed. In order to correlate this ORF with a carboxysome peptide, the ORF was overexpressed in E. coli by cloning it into pProExHTb, a prokaryotic expression vector containing an E. coli ribosome binding site. When antibodies raised against the recombinant protein were used to probe an immunoblot containing carboxysome peptides, a 60-kDa peptide was recognized. The peptide was subsequently named CsoS3. CsoS3 is a minor component of the carboxysome; a peptide of this size is commonly not observed or is very faint on Coomassie blue-stained SDS-polyacrylamide gels of purified carboxysomes. Immunogold labeling established CsoS3 to be a component of the carboxysome shell. PMID- 10816047 TI - Effect of nystatin on the release of glycerol from salt-stressed cells of the salt-tolerant yeast Zygosaccharomyces rouxii. AB - The polyene antibiotic nystatin, which affects fungal membrane permeability, inhibited the growth of Zygosaccharomyces rouxii grown in medium containing 15% (w/v) NaCl, whereas yeast grown in medium without NaCl were only slightly inhibited. Nystatin caused salt-stressed cells to release large amounts of glycerol and inhibited their growth, but amino acids and materials with an absorbance at 260 nm were not released from the cells. The leakage was increased by the addition of glucose, and more than 90% of the intracellular glycerol was released into the medium during a 2-h incubation with 0.11 microM nystatin and 2% (w/v) glucose. Glycerol was indispensable for the growth of Z. rouxii grown in culture medium containing 15% NaCl. PMID- 10816048 TI - Initial steps in the fermentation of 3-hydroxybenzoate by Sporotomaculum hydroxybenzoicum. AB - The anaerobic bacterium Sporotomaculum hydroxybenzoicum ferments 3 hydroxybenzoate to acetate, butyrate, and CO2. 3-Hydroxybenzoate was activated to 3-hydroxybenzoyl-CoA in a CoA-transferase reaction with acetyl-CoA or butyryl-CoA as CoA donors. 3-Hydroxybenzoyl-CoA was reductively dehydroxylated, forming benzoyl-CoA. This reaction was measured in cell-free extracts with cob(I)alamin as low-potential electron donor. No evidence was obtained that cob(I)alamin is the physiological electron donor; however, inhibitor studies indicated involvement of a strong nucleophile in the reaction. Benzoate was degraded by dense cell suspensions without a lag phase until an in situ deltaG' value <-25 kJ mol(-1) was reached. Benzoyl-CoA reductase was not detected. Enzyme activities for all reaction steps from glutaryl-CoA to butyryl-CoA, and ATP formation via acetate kinase were detected in cell-free extracts. Glutaconyl-CoA decarboxylase is likely to act as a primary sodium ion pump. PMID- 10816049 TI - Heterogeneity of the genus Myrothecium as revealed by cell wall polysaccharides. AB - The polysaccharides obtained from the alkali-extractable, water-soluble fraction (F1SS) from the cell wall of Myrothecium verrucaria and Myrothecium atroviride were shown to be composed of beta-(1-->6)-galactofuranose fully substituted at O 2 by terminal residues of alpha-glucopyranose and alpha-glucuronic acid. Glucuronic acid was substituted at O-4 by glucopyranose in the Myrothecium species M. inundatum, M. setiramosum, M. prestonii, M. tongaense and M. roridum. The acidic polysaccharides from Phaeostilbella atra (=Myrothecium atrum) and Myrothecium gramineum lacked the backbone of 2,6 di-O-substituted galactofuranose and contained a high amount of O-5-substituted beta-galactofuranose. The structures of the polysaccharides isolated from Myrothecium cinctum and Myrothecium penicilloides were unrelated to each other and to the polysaccharides from the other species analysed. The usefulness of these polysaccharides as a characteristic for delimitation of the genus Myrothecium is discussed. PMID- 10816050 TI - Identification of an L-dopa decarboxylase gene from Sorangium cellulosum So ce90. AB - During a screening program intended to identify genes encoding enzymes typical for secondary metabolism in Sorangium cellulosum So ce90, an aromatic amino acid decarboxylase gene (ddc) was detected. Expression of ddc in Escherichia coli and subsequent enzyme assays with cell-free extracts confirmed the proposed function derived from amino acid sequence comparisons. In contrast to other aromatic amino acid decarboxylases of eukaryotic origin, the S. cellulosum Ddc converted only L dihydroxy phenylalanine. This is the first report of a gene encoding an L dihydroxy phenylalanine decarboxylase in bacteria. PMID- 10816051 TI - A comparison of solid and liquid media for resuscitation of starvation- and low temperature-induced nonculturable cells of Aeromonas hydrophila. AB - Like many other gram-negative bacteria, starved cells of Aeromonas hydrophila can be induced into a viable but nonculturable (VBNC) state by incubation at low temperature, as shown here by using various bacterial enumeration methods. Starved A. hydrophila strain HR7 cells at 4 degrees C reached the nonculturable stage in about 45 days. The cells were resuscitated by either a solid medium resuscitation method, using solid agar amended with H2O2-degrading agents, catalase or sodium pyruvate, or a liquid medium resuscitation method, by incubating nonculturable cells in liquid media containing these compounds before spreading onto plates. The liquid medium resuscitation method using catalase resulted in nearly complete recovery of nonculturable cells. PMID- 10816052 TI - Highly tumorigenic Agrobacterium tumefaciens strain from crown gall tumors of chrysanthemum. AB - A wild-type Agrobacterium tumefaciens strain CNI5 isolated from crown gall of chrysanthemum (Dendranthema grandiflora Tzvelev) was characterized. Strain CNI5 was classified into biovar 1, based on physiological and biochemical characteristics, and was resistant to ampicillin. Strain CNI5 induced tumors at a higher frequency and on a larger area of explants in most tested plant species, especially in chrysanthemum cultivars, than the octopine-type strain C58C1cmr (pTiB6S3). Agropine and mannopine were detected in tumors induced by strain CNI5 and were specifically catabolized by this strain. Strain CNI5 harbored five plasmids including one plasmid that shared sequence similarity to TL-DNA of the octopine-type Ti plasmid and four cryptic plasmids. PMID- 10816053 TI - Implications of the FRISCII trial: initial medical stabilization followed by an early invasive approach is the preferred strategy in unstable coronary syndromes. PMID- 10816054 TI - Off-pump vs on-pump surgery. A review. PMID- 10816055 TI - Infections and atherosclerosis. AB - More than a century ago, inflammation and infection were considered to have atherogenic effects. During last century, however, this hypothesis was completely abandoned, and the old idea that coronary heart disease (CHD) possibly has an infectious etiology has only re-emerged in recent years. Both viral and bacterial pathogens have been proposed to be associated with the inflammatory changes found in atherosclerosis. Herpes group viruses, especially cytomegalovirus (CMV) and herpes simplex virus type 1 (HSV1), have been associated with both atherosclerosis and restenosis. Helicobacter pylori and dental infections have also been linked to CHD, but the evidence is strongest for a respiratory tract bacterium, Chlamydia pneumoniae. The association was originally found in seroepidemiological studies, but the presence of organisms in atherosclerotic lesions, the first animal studies and preliminary successful intervention trials with antibiotics suggest that C. pneumoniae may have a pathogenetic role in the disease. The causal relationship has not yet been proven, but ongoing large intervention trials and research on pathogenetic mechanisms may lead to the use of antimicrobial agents in the treatment of CHD in the future. PMID- 10816056 TI - Cardiac endothelin-like immunoreactivity and preproendothelin-1 mRNA expression in human heart failure. AB - Endothelin (ET) induces hypertrophy of cardiomyocytes and increases synthesis of collagen in vitro. Interestingly, these features are hallmarks of the cardiac remodeling taking place in heart failure. The aim of the present study was to examine cardiac ET peptide and preproET-1 mRNA synthesis in human heart failure. Cardiac tissue was obtained from 11 patients with end-stage heart failure undergoing orthothopic heart transplantation (NYHA III-IV). Cardiac tissue from nine organ donors served as controls. The specimens were examined by immunohistochemistry and mRNA slot blot analyses. Significantly stronger ET-1 like immunoreactivity (ET-1-ir) was seen in the left atrial myocardium of failing hearts compared to the left atrial myocardium of donor hearts. Within each heart, the epicardium showed the strongest ET-1-ir. Left ventricular preproET-1 mRNA expression in the entire group of patients did not differ significantly from that of donor hearts. However, hypertrophic obstructive cardiomyopathy may be associated with a twofold increase in left ventricular preproET-1 mRNA. We report an increase in cardiac ET peptide in human heart failure. PMID- 10816057 TI - Effects of L-arginine on vascular smooth muscle cell proliferation and apoptosis after balloon injury. AB - Nitric oxide (NO) inhibits neointimal formation in experimental models of restenosis, but the mechanisms have not been fully elucidated. This study examined whether the beneficial effect of L-arginine, the physiological NO precursor, was associated with alteration of the apoptotic and proliferative activities of vascular smooth muscle cells (VSMCs) in the vessel wall after arterial injury. Balloon injury was performed in the rat carotid-artery injury model. Rats were treated with L-arginine (2.25% in the drinking water) or normal drinking water, and sacrificed at 1, 2 and 14 days postinjury. Apoptosis was assessed by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL), and proliferation by proliferating cell nuclear antigen (PCNA) immunohistochemistry. Treatment with L-arginine increased the luminal area at 14 days postinjury (0.26 +/- 0.03 mm2 vs 0.14 +/- 0.04 mm2; p < 0.05), and this effect was attributable to a reduction in neointimal formation (0.11 +/- 0.03 mm2 vs 0.23 +/- 0.04 mm2; p < 0.05), while L-arginine did not affect vascular remodeling, as indicated by the total vessel area. The decreased neointimal area at 14 days after balloon injury contained a reduced percentage of TUNEL positive (0.1 +/- 0.1% vs 2.0 +/- 0.6%; p < 0.05), and PCNA positive (13.0 +/- 2.6% vs 27.2 +/- 5.9%; p < 0.05) nuclei, respectively. L-arginine did not influence the apoptotic or proliferative activities of VSMCs at earlier time points postinjury. The favourable effect of L-arginine in the rat model of arterial injury is associated with inhibition of VSMC proliferative activity in the vessel wall and is not explained by increased VSMC apoptosis. PMID- 10816059 TI - Perioperative stroke in coronary artery bypass surgery. Analysis of risk factors. AB - The incidence of neurologic complications after coronary bypass surgery is steadily rising as older and sicker patients are increasingly being treated. To identify patients requiring special attention, we reviewed the records in 2263 consecutive cases of first-time coronary artery bypass grafting in 1993-1995, in order to find predictive factors for stroke. Significant factors in univariate analysis were patient age, peripheral vascular disease, cerebrovascular disease, renal failure (defined as serum creatinine > or = 150 micromol/l), aneurysmal disease of the abdominal aorta, stenosis of the left main coronary artery, urgent or emergency operation, NYHA class, cardiopulmonary bypass time, number of aortic anastomoses, intraoperatively detected loose or calcified atheromatosis of the ascending aorta, left ventricular venting, intra-aortic balloon counterpulsation, cardiac complications necessitating early reoperation, and perioperative myocardial infarction. In a multivariate analysis, age, renal failure, cerebrovascular disease, peripheral vascular disease, NYHA class, number of aortic anastomoses, perioperative myocardial infarction and intraoperatively detected loose atheromatosis of the ascending aorta remained significant. PMID- 10816058 TI - Heparin-coated cardiopulmonary bypass circuits reduce circulating complement factors and interleukin-6 in paediatric heart surgery. AB - Children are sensitive to the inflammatory side effects of cardiopulmonary bypass (CPB). Our intention was to investigate if the biocompatibility benefits of heparin-coated CPB circuits apply to children. In 20 operations, 19 children were randomized to heparin-coated (group HC, n = 10) or standard (group C, n = 10) bypass circuits. Plasma levels of acute phase reactants, interleukins, granulocytic proteins and complement factors were measured. All were significantly elevated after CPB. Levels of complement factor C3a (851 (791 959)ng/ml [median with quartiles] in group C, 497 (476-573)ng/ml in group HC, p < 0.001), Terminal Complement Complex (114 (71-130) AU/ml in group C, 35.5 (28.9 51.4) AU/ml in group HC, p < 0.001), and interleukin-6 (570 (203-743) pg/ml in group C, 168 (111-206)pg/ml in group HC, p = 0.005), were significantly reduced in group HC. Heparin-coated CPB circuits improve the biocompatibility of CPB during heart surgery in the paediatric patient population, as reflected by significantly reduced levels of circulating complement factors and interleukin-6. PMID- 10816060 TI - Preoperative and postoperative response to inhaled nitric oxide. AB - The preoperative dose response to inhaled nitric oxide (NO) was compared with the need for and response to NO after cardiac surgery in patients with congenital heart defect and secondary pulmonary hypertension. In a preoperative vasodilator test with inhaled NO 20, 40 and 80 ppm and oxygen, mean pulmonary artery pressure (PAP) was at least 40 mmHg and/or the pulmonary vascular resistance index (PVRI) 4 Wood units. Preoperatively, NO 40 ppm and FiO2 0.9 reduced systolic pulmonary/systemic arterial pressure (PAPs/SAPs) from 0.89 (SD 0.10) to 0.80 (0.18) and pulmonary/systemic vascular resistance (PVR/SVR) from 0.26 (0.13) to 0.13 (0.08). Haemodynamic assessment was repeated in 11 patients postoperatively. NO treatment was started if PAPs/SAPs rose to 0.8 or the pulmonary oximetry fell below 40%. Postoperatively, eight of 11 patients, including 6 patients with Down's syndrome, needed NO. PAPs/SAPs decreased more than preoperatively: 48.5% vs 11.2, p = 0.0045. Pulmonary oximetry increased by 15.7%, p = 0.02. The degree of preoperative response to NO did not differ between the patients with postoperative pulmonary hypertension and the other children. Patients with early pulmonary hypertensive crisis (first 24 h; n = 6) had a higher PVRI (7.6 vs 4.4 Um2; p = 0.003) and PVR/SVR (0.34 VS 0.17; p = 0.02) preoperatively. Two patients died in pulmonary hypertensive crisis. PMID- 10816061 TI - Baseline platelet aggregation and major receptor expression predict subsequent activity following thrombolysis for acute myocardial infarction. AB - The early identification of patients with heightened platelet activity for aggressive antiplatelet regimens represents a critical clinical issue in acute myocardial infarction (AMI) therapy. We sought to determine whether the degree of pre-reperfusion platelet function is related to subsequent activity following thrombolysis. Platelets were investigated at baseline and at 24 h following thrombolytic therapy by aggregometry, and flow cytometry in 19 AMI patients enrolled in the GUSTO-III trial. Regression analysis revealed a significant correlation for 5 microM ADP (r2 = 0.529), 10 microM ADP (r2 = 0.445), thrombin (r2 = 0.226), collagen (r2 = 0.568), and ristocetin-induced aggregation (r2 = 0.964). Platelet expression linearly correlated for GPIIb/IIIa (r2 = 0.337), P selectin (r = 0.817), PECAM-1 (r2 = 0.586), and the vitronectin receptor (r2 = 0.634). The data suggest that the baseline characteristics predict future platelet activity, and may prospectively identify patients who will benefit most from antiplatelet strategies after coronary thrombolysis. PMID- 10816062 TI - Randomised training after myocardial infarction: short and long-term effects of exercise training after myocardial infarction in patients on beta-blocker treatment. A randomized, controlled study. AB - In this study we assessed the short- and long-term effects of 4-weeks of exercise training (MI) soon after myocardial infarction in patients on beta-blocker treatment. Thirty-seven male patients < or = 65 years of age were included in the study, 19 of them randomized to exercise training (ET) and 18 to a control group (Ctr). Cumulated work (CW), calculated in kiloJoules (kJ), was recorded before immediately after the intervention period and again six months after the MI. In the short term the mean (SD) CW increased by 22% (from 65(20) to 79(25) kJ) in the ET group, compared with no change in the Ctr patients (65(24) vs 65(21) kJ) (p = 0.009). At late follow-up CW was 14% above baseline in the ET patients (65(20) vs 74(20) kJ) p = 0.036, compared with only 6% in the 15 Ctr patients who were still available for follow-up (68(24) vs 72(29) kJ), but without a significant between-group difference. In post-MI patients on beta-blocker treatment, and with a high baseline exercise capacity, physical training improved exercise capacity in the short term, but there was no significant between-group difference at long-term follow-up. PMID- 10816063 TI - Limitation of physical activity, dyspnoea and chest pain before and two years after coronary artery bypass grafting in relation to preoperative ejection fraction. AB - To investigate the relationships between limitation of physical activity and dyspnoea and chest pain before and 2 years after coronary artery bypass grafting (CABG) and preoperative left ventricular ejection fraction (LVEF), questionnaires were issued to all patients from Western Sweden who underwent CABG during 1988 1991. The analysis comprised 985 patients. Physical activity improved significantly after CABG regardless of the preoperative LVEF. No significant association was found between LVEF and degree of limitation of physical activity before or after surgery. Dyspnoea and chest pain improved markedly, irrespective of LVEF. There was significant association between freedom from dyspnoea and LVEF preoperatively (less dyspnoea with higher LVEF), but not after CABG. The frequency of chest-pain attacks was not related to LVEF, before or after the operation. Thus physical activity, dyspnoea and chest pain improved in the 2 years after CABG irrespective of preoperative LVEF. Absence of dyspnoea was related to LVEF before, but not after surgery, and there was no association between preoperative LVEF and frequency of anginal attacks before or after CABG. PMID- 10816064 TI - Current smokers develop more posterior myocardial infarctions probably due to increased tendency to thrombosis. AB - This investigation was carried out to determine whether smokers developed smaller infarcts as assessed by peak enzyme levels and also to what extent smoking could modify infarct localization. The study included 753 patients, of whom 351 had no history of previous coronary heart disease (CHD) (angina pectoris and/or myocardial infarction (MI)). The investigation was designed as an exposed (smoking) versus non-exposed (non-smoking) cohort study. Outcome was infarct size, posterior versus non-posterior MI and non-Q-wave versus Q-wave infarcts. In the total cohort of patients, 312 (41%) were smokers, the corresponding number in the restricted cohort of patients without a previous CHD (CHD-0-pts) was 169 (48%). Smokers were younger than non-smokers, and more of them were males. It was found that infarct size was similar in smokers and in non-smokers (crude and adjusted effects). Crude effects showed that smokers developed significantly more posterior infarcts than non-smokers; odds ratio (OR) for developing a posterior MI was 1.95 (2p < 0.001) (all patients) and 2.34 (2P < 0.001) (CHD-0-pts), respectively. After adjusting for confounders (logistic regression model), OR in the two groups was 1.24 (2p = 0.256) and 1.95 (2p = 0.01), respectively. The study shows that current smokers were younger, and indicates that in those without a previous CHD, significantly more of them developed a posterior MI. PMID- 10816065 TI - Atrial septal defect in adults. Thirty-eight-year follow-up of a surgically and a conservatively managed group. AB - This report describes the long-term (up to 38 years) outcome, in terms of mortality and cardiovascular morbidity, in a non-randomized clinical observation study of a surgically and a conservatively managed group of adult patients with shunt lesions at atrial level. Haemodynamic status was assessed at baseline and at repeat examination. Twelve patients underwent surgical repair of the lesion between these two heart catheterizations, resulting in a marked reduction in heart size and right ventricular systolic pressure. These changes were associated with improvement in functional class and a trend towards less risk of cerebrovascular incidents, but not atrial fibrillation, during follow-up. Among 12 patients not operated on between catheterizations, symptomatic deterioration was common, often necessitating later surgical repair. The results support the assumption that early surgery should be recommended for adults with a haemodynamically significant lesion, to reduce the risk of mortality and prevent symptomatic deterioration. PMID- 10816066 TI - Percutaneous transradial coronary angiography and angioplasty in patients with occlusive atherosclerotic iliofemoral disease. AB - Not all coronary angiograms can be acquired through the femoral route. The transradial catheterisation procedure in patients with occlusive atherosclerotic iliofemoral disease is described. Transfemoral left-sided cardiac catheterisation was performed in approximately 99.5% of patients referred for coronary angiography, while out of 48 patients in whom transfemoral access was impossible, transradial coronary angiography was successful in 37. With the exception of one, all patients with coronary artery disease had lesions of the right coronary artery, more than 70% had multivessel disease and 14% had stenosis of the left main coronary artery. Ten patients had angioplasty performed during the same procedure. Complications occurred in 5 out of 39 cases, 2 (5%) of these were severe. Although the femoral route was used in more than 99% of an unselected population referred for coronary angiography, it was found that transradial angiography and angioplasty can be performed in patients with occlusive atherosclerotic iliofemoral disease with considerable success and an acceptable complication rate. PMID- 10816067 TI - Comparison of two automatic staplers for closing the bronchial stump after pulmonary resection. AB - The incidence of complications following pulmonary resection using an automatic stapler was studied in 348 consecutive patients. A parallel-jaw stapler with two staple lines was used in group A (133 males, 101 females, mean age 66 years) between 1990 and 1995, and a hinged-jaw stapler with three staple lines in group B (70 males, 44 females, mean age 67 years) in 1995-1997. The incidence of bronchopleural fistula was 0.4% (1 patient) in group A and nil in group B. The incidence of intraoperative air leak requiring interrupted suture closure was 4.7% (11 patients) in group A and nil in group B (p < 0.05). Postoperative bronchopleural fistula did not develop in any of the cases, and there was no mortality. For management of the bronchial stump after pulmonary resection, the newer device firing three rows of staples is superior to the two-row device. PMID- 10816068 TI - Strategy for 3-D computed tomography diagnosis and treatment of small pulmonary arteriovenous fistula. AB - In a patient with left hemianopsia, computed tomography (CT) and magnetic resonance imaging revealed an intracranial space-occupying lesion in the right occipital region. A cerebral abscess was removed at craniotomy. Enhanced pulmonary CT showed a small coin lesion in the peripheral lingula. and at 3-D CT two nidi with feeding arteries and draining veins were seen, indicating pulmonary arteriovenous fistula. Lingular segmentectomy was performed. PMID- 10816069 TI - Successful closure of postpneumonectomy bronchopleural fistulae by latissimus dorsi island flap and closed-chest irrigation. AB - Bronchopleural fistula after pneumonectomy is a dreaded complication, because of its high morbidity and mortality. We describe a successful technique using closed chest irrigation of the pleural cavity continued with transposition of a latissimus dorsi island flap. PMID- 10816070 TI - The drug cerebrolysin and its peptide fraction E021 increase the abundance of the blood-brain barrier GLUT1 glucose transporter in brains of young and old rats. AB - The brain-derived peptidergic drug Cerebrolysin has been found to support the survival of neurons in vitro and in vivo. In the present study, we investigated the effects of Cerebrolysin and its peptide preparation E021 on spatial learning and memory, as well as on the abundance of the blood-brain barrier GLUT1 glucose transporter (GLUT1) in 2-month-old and 24-month-old rats. Young rats were treated with the drugs or saline (2.5 ml/kg/day) daily on postnatal days 1-7, and old rats for 19 consecutive days. For behavioural testing the Morris water maze was used. The abundance of GLUT1 was determined in brain slices by immunocytochemistry. Quantification of the density of the GLUT1 immunostaining was performed using light microscopy and a computerised image analysing system. All drug-treated rats, young and old, exhibit shorter escape latencies in the water maze, on all testing days (p < 0.01), indicating improved cognitive performance. Immunohistochemical data show an age-related decrease of the density of GLUTI (p < 0.05). In young animals, the administration of the drugs led to an increase of the abundance of GLUT1 in all experimental groups (p < 0.01). In old rats, the treatment with Cerebrolysin, but not with E021, resulted in an increase in the immunoreactive GLUT1 (p < 0.01). The elevated abundance of GLUT1 after the administration of both peptidergic substances might be supportive for the cognitive effects of this drug, by causing an improved nutritional supply of glucose to the neurons. PMID- 10816071 TI - Two peptidergic drugs increase the synaptophysin immunoreactivity in brains of 6 week-old rats. AB - An increase of synaptic density has been found in the hippocampus, the dendate gyrus and in the entorhinal cortex of 6-week-old rats after 7 days of treatment with the peptidergic drug Cerebrolysin, its peptide preparation E021 and the diluted peptide preparation E021dil. Rats received drugs on postnatal days 1-7 (2.5 ml/kg, each day). Controls received saline. The animals were sacrificed on days 42-48 of their life, after they had undergone behavioural testing in a Morris water maze. Slices of brain were stained immunohistochemically with anti synaptophysin, a specific marker of presynaptic terminals. The synaptophysin immunoreactivity of presynaptic terminals was quantified using light microscopy and a computerised image analysis system. Our results showed that rats benefit from the treatment with both drugs. A significant increase in the number of synaptophysin-immunoreactive presynaptic terminals was found in the entorhinal cortex and the hippocampal subfields CA1, CA2, CA3 stratum radiatum and CA3 stratum lucidum. The increased immunoreactive presynaptic terminals found in the present study are in accordance with the positive effects of the drugs on spatial learning and memory in young rats (Gschanes & Windisch 1999). PMID- 10816072 TI - Immunohistochemical study on Fas and Fas ligand in skin wound healing. AB - An immunohistochemical study on the expression of Fas and Fas ligand (Fas L) was performed in order to examine the role of apoptosis through Fas-Fas L in mouse skin wound healing. After a 1-cm-long incision in the central dorsum skin, mice were sacrificed at intervals ranging from 0.5 to 240 h, followed by the sampling of wound margin. The expression of Fas and Fas L in the wound margins and in uninjured skin controls was studied using frozen sections. In uninjured skin controls, a very weak expression of Fas and Fas L was detected immunohistochemically in hair follicles, sebaceous glands and epidermal cells. In wounded specimens, polymorphonuclear cells and inflammatory mononuclear ones (round-shaped and spindle-shaped types) were evident. A single immunostaining showed that Fas or Fas L was detectable in inflammatory mononuclear cells involved in the skin wound healing process. Double immunostaining for Fas and Fas L revealed that inflammatory mononuclear cells co-expressed both antigens. In situ TUNEL combined with immunostaining showed that the inflammatory mononuclear cells expressing Fas or Fas L and the polymorphonuclear cells were TUNEL-stained, although neither Fas nor Fas L was detected in the polymorphonuclear cells. The number of TUNEL-positive, inflammatory mononuclear cells expressing Fas or Fas L per 0.01 x 0.01 cm2 was counted. The average number of 10 randomly selected microscope fields reached a peak at the fibro-proliferative phase of wound healing. These results indicate that apoptosis through Fas and Fas L may play an important role for reducing the cellularity during skin wound healing in mice. PMID- 10816073 TI - Immunohistochemical analysis of DNA mismatch repair enzyme hMSH-2 in normal human skin and basal cell carcinomas. AB - We have analysed the expression and distribution of the DNA mismatch repair enzyme hMSH-2 in normal skin and basal cell carcinomas. hMSH-2 protein was investigated immunohistochemically (normal human skin: n = 10; basal cell carcinomas: n = 16) on frozen sections using a highly sensitive streptavidin peroxidase technique and a specific mouse monoclonal antibody (clone FE11). In normal human skin, we found nuclear immunoreactivity for hMSH-2 in epidermal keratinocytes of the basal and first 1-3 suprabasal cell layers. All basal cell carcinomas analysed revealed strong nuclear imunoreactivity that was pronounced in peripheral tumour cells and cells of the palisade. Expression of hMSH-2 protein was consistently and strongly upregulated in tumour cells of the carcinomas as compared to adjacent unaffected epidermis or epidermis of normal human skin. Twelve of the sixteen carcinomas analysed revealed no visual correlation in comparing the labelling patterns for hMSH-2 with the labelling pattern for the proliferation marker Ki-67. Our findings indicate that (a) hMSH-2 is expressed in human epidermal keratinocytes, predominantly in lower cell layers of the viable epidermis; (b) expression of hMSH-2 protein is strongly upregulated in basal cell carcinomas as compared to unaffected epidermis; (c) the level of hMSH-2 proteins in the carcinomas is not exclusively regulated by the proliferative activity of these tumour cells; (d) inactivating mutations of the hMSH-2 gene may in the carcinomas not be involved in the carcinogenesis or microsatellite instability secondary to replication errors; (e) expression of hMSH-2 may be of importance for the genetic stability of basal cell carcinomas in vivo. PMID- 10816074 TI - Rapid and sensitive detection of enhanced green fluorescent protein expression in paraffin sections by confocal laser scanning microscopy. AB - Various forms of green fluorescent protein (GFP) have become important reporters of gene transfer and expression after transfection or infection of cells in cell culture. Frequently, molecular biological assays (Northern blots, PCR) are applied to detect reporter gene expression in target organs. However, these methods are not suitable for evaluation of tissue- or cell-specific expression which would be of great interest especially in case of using tissue-specific promoters. Therefore, organs of transgenic mice with the enhanced green fluorescent protein (EGFP) gene under control of the cytomegalovirus (CMV) promoter were processed for histology by formaldehyde fixation and embedding in paraffin. Sections were deparaffinized, mounted and evaluated for fluorescence in a confocal laser scanning microscope. This method combines the advantages of direct exploitation of tissue sections without further staining procedures with evaluable tissue-, cell-, and even subcellular-specific distribution patterns of EGFP expression in tissues. Results obtained by direct evaluation of EGFP fluorescence in paraffin sections were confirmed by immunohistochemical staining with anti-EGFP. In the present report, we demonstrate that application of confocal microscopy on routinely processed histological preparations is very suitable for determining gene transfer efficiency and promotor activities. PMID- 10816075 TI - Cepaea hortensis agglutinin-I, specific for O-glycosidically linked sialic acids, selectively labels endothelial cells of distinct vascular beds. AB - The lectin Cepaea hortensis agglutinin-I (CHA-I) binds to O-glycosidically linked sialic acids with previously characterized specificity. Employing histochemistry, we demonstrate that CHA-I is a useful probe for detecting sialic acids in formalin-fixed human tissues in a specific manner. It stains the endothelium of arteries and veins in all tissues examined, and labels the capillaries in distinct vascular beds including the brain, colon, thyroid, pituitary, and adrenal. By contrast, the endothelial sinusoids in the liver, spleen, and bone marrow remained unstained. The staining pattern of CHA-I overlaps with the distribution of the sialomucin and L-selectin ligand podocalyxin, which includes positivity of podocytes and interstitial but not glomerular capillaries. CHA-I positive epithelial structures were found in the lung, liver and kidney. Colon carcinoma cells were labelled with CHA-I but not haemangiosarcomas. In summary, CHA-I is a useful tool for detecting O-glycosidically linked sialic acids in formalin-fixed tissues, and a potentially powerful tool for the isolation and characterization of unknown sialomucins in normal and eventually in diseased tissues. PMID- 10816076 TI - Application of the critical molar concentration concept to heat-mediated antigen retrieval in immunohistochemistry. AB - Previously we have applied Scott's critical molar concentration concept to show that divalent cations, especially Mg2+ may be used to measure the affinity of a known monoclonal antibody for its antigen. In this paper we report the application of this same procedure to a study of a series of antigens (three globular proteins and three intermediate filaments). The concept was applied to samples without any previous treatment or after the application of heat-mediated antigen retrieval (using a pressure cooker). Our findings suggest that heat mediated antigen retrieval sets free protein side-chain(s) that have been masked by formaldehyde fixation. This is reflected in a higher affinity of the antibody for the antigen in question. PMID- 10816077 TI - Programmed cell death in 2',3'-dideoxycytidine-resistant human monoblastoid U937 cells. AB - 2',3'-Dideoxycytidine is a powerful in vitro inhibitor of human immunodeficiency virus and is currently used in the treatment of acquired immunodeficiency syndrome. A long-term exposure of U937 monoblastoid cells to dideoxycytidine induces the selection of drug-resistant cells (U937-R). In previous studies, we investigated some important biochemical properties and functional activities, such as basal respiration, protein kinase C activity, superoxide anion release, and the level of reduced glutathione, which were found to be higher in the drug resistant cell line, compared to the parental one. In the present study, we evaluated the response of the two cell lines to the induction of apoptosis by treatment with staurosporine and okadaic acid, which interfere with the protein kinase and phosphatase pathways, respectively. Moreover, knowing that GSH plays a crucial role in the regulation of nitric oxide-dependent apoptosis, U937-R and parental lines have been treated with SIN-1, which is known to generate significant amounts of O2 and nitric oxide. Resistant and parental cells have been analysed by light and electron microscopy and agarose gel electrophoresis of isolated DNA has been performed. The obtained results demonstrate a different susceptibility of U937-R cell line to apoptosis induced with the three triggers. U937-R cells show more advanced apoptotic features if compared with parental cells, after staurosporine treatment. Differently, the okadaic acid does not induce a different behaviour in the two models. On the contrary, the agent SIN-1 determines an increased number of apoptotic cells in the U937 line. The results suggest that a higher level of protein kinase C and glutathione could prevent programmed cell death in U937-R. PMID- 10816078 TI - Localization of galanin-like immunoreactive structures in the brain of the Senegalese sole, Solea senegalensis. AB - The distribution of galanin-like immunoreactive structures was studied in the brain of the Senegalese sole, Solea senegalensis, using immunohistochemical methods. Periventricular immunoreactive cell bodies were observed in the rostral pole of the preoptic recess, within the pars parvocellularis of the nucleus preopticus parvocellularis. Another galanin-immunoreactive cell population was observed more caudal in the ventromedial hypothalamus, along the medial evaginations of the lateral recess. These cells appear within the cytoarchitectonic limits of the nucleus recessus lateralis pars ventralis. We found an extensive presence of galanin-immunoreactive fibres throughout the entire brain, although the most massive network of fibres was observed in the caudal olfactory bulbs, ventral telencephalon, preoptic area and around diencephalic ventricular recesses. Also, the hypophysis, ventricular mesencephalic area, median reticular formation and viscerosensory rhombencephalon displayed important plexuses of galanin-immunoreactive axons. The widespread distribution of these immunoreactive structures in the brain and pituitary of the Senegalese sole suggests an important role for galanin in neuroendocrine regulation of brain and adenohypophyseal functions. PMID- 10816079 TI - Integrin-associated protein (IAP)-deficient mice are less susceptible to developing Staphylococcus aureus-induced arthritis. AB - The integrin-associated protein (IAP) has been shown to function in a signaling complex with beta3 integrins, influencing the migration of phagocytic cells into inflamed tissues. We have previously shown that gene-targeted mice deficient for IAP succumbed to peritonitis when inoculated with gram-negative bacteria. The aim of this study was to assess the role of IAP in our recently established model of haematogenously induced Staphylococcus aureus septicaemia and arthritis. In this model, neutrophils play a crucial role in the early phase of the infection. Mice lacking IAP and congenic controls were intravenously inoculated with S. aureus LS 1. The IAP-/- mice were resistant to developing clinical signs of arthritis compared with their IAP-expressing littermates. The clinical findings were corroborated by histopathological evaluation indicating that the IAP-/- mice had less cartilage and bone destruction in the joints. We believe that a delayed migration of leukocytes into the joints of mice lacking IAP expression leads to decreased susceptibility to develop S. aureus-induced arthritis. PMID- 10816080 TI - Cytolytic T-cell responses to human dendritic cells and macrophages infected with Mycobacterium bovis BCG and recombinant BCG secreting listeriolysin. AB - Cytolytic T-cell responses from 63 normal blood donors were monitored in a Mycobacterium bovis BCG infection system in vitro. We wanted to know whether cultured dendritic cells were capable of potentiating the cytolytic T-cell responses to M. bovis BCG. Infected cultured dendritic cells were up to ten times more effective antigen-presenting cells than macrophages in proliferative assays, while cytolytic T-cell induction did not differ significantly between dendritic cells and macrophages. Separated CD4+ and CD8+ T-cell subsets contributed equally to lysis of infected targets. Experiments comparing wild-type M. bovis BCG strain with two new recombinant M. bovis BCG strains secreting listeriolysin revealed statistically significant higher maximal lysis values for recombinant M. bovis BCG. We conclude from our in vitro infection system with mycobacteria that dendritic cells are superior to macrophages in proliferative assays but equal to macrophages in their ability to induce cytolytic T-cell responses. Moreover, our data suggest that recombinant M. bovis BCG vaccine strains secreting listeriolysin improve cytolytic T-cell responses. PMID- 10816081 TI - Experimental conditions that increase the production of HIV-1 by monocyte-derived macrophages: use of collagen matrix. AB - Monocyte-derived macrophages (MDMs) from healthy blood donors were isolated by adherence to tissue culture-treated plasticware. They were cultured in vitro in medium supplemented with human serum and recombinant GM-CSF, then infected with the macrophage-tropic prototype strain HIV-1-PAR. Virus production was quantitated at various times after infection by measuring reverse transcriptase concentration in cell-free tissue culture supernatant fluids, using a sensitive nonradioactive assay. Virus production was significantly increased by culturing MDMs on plasticware previously coated with collagen 1. The increase in virus production was dependent upon collagen 1 concentration, with maximal value being encountered after coating with 1.5 microg/cm2. These results indicate that the sensitivity of peripheral macrophages to HIV-1 infection might be influenced by contact-dependent interactions involving components of the extracellular matrix that take place during the process of monocyte extravasation and migration. PMID- 10816082 TI - Increase in CD3+ CD4- T lymphocytes in patients with AIDS and disseminated Mycobacterium avium-intracellulare complex infection: a prospective study. GECSA. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine. AB - In a retrospective study, an increase in double-negative (CD3+ CD4- CD8-) (DN) T lymphocytes has been shown to be an independent predictor of disseminated Mycobacterium avium complex (D.MAC) infection in patients with less than 100 CD4+ T cells per mm3. To better characterize this cell expansion, a prospective study was designed. From July 1995 to April 1997, 206 HIV-infected patients with less than 100 CD4+ T cells per mm3 were prospectively followed up and immunophenotyped. The median followup was 1.1 year (+/-0.5 year), and 14 new D.MAC infections were diagnosed among 84 first AIDS-defining events. In univariate and multivariate analyses, D.MAC infections were the only opportunistic infection with a significant increase in DN T-cell percentage (median = 6.6; range = 1.7 to 24.5, P = 0.004) compared with patients without any opportunistic infection. This alteration in T-lymphocyte count could constitute a predictor for D.MAC infection in clinical practice. PMID- 10816083 TI - A review of oral vaccination with transgenic vegetables. AB - Mucosal immunization of the gastrointestinal tract is an effective way to stimulate local and systemic immune responses. Oral vaccines must be formulated in such a way that antigens are protected as they pass through the adverse environment of the stomach and are delivered to the mucosal inductive sites. Vaccine antigens cloned into edible transgenic plants are a promising new delivery system for oral vaccines. Such vaccines could be safe, inexpensive, and multicomponent. PMID- 10816084 TI - Complement-resistance mechanisms of bacteria. AB - Despite more than a century of parallel research on bacteria and the complement system, relatively little is known of the mechanisms whereby pathogenic bacteria can escape complement-related opsonophagocytosis and direct killing. It is likely that pathogenicity in bacteria has arisen more accidentally than in viruses, and on the basis of selection from natural mutants rather than by outright stealing or copying of genetic codes from the host. In this review we will discuss complement resistance as one of the features that makes a bacterium a pathogen. PMID- 10816085 TI - Antimicrobial resistance in Staphylococcus aureus. AB - Recognized since 1883 as a common cause of infection, Staphylococcus aureus' preantimicrobial-era bacteremia mortality rate was 82%. The mortality of that era threatens to return as evidence of growing vancomycin resistance undermines the utility of vancomycin therapy. Successful treatment of S. aureus infections requires knowledge of its antimicrobial resistance capacity. PMID- 10816086 TI - Host-parasite interaction in bovine infection with Tritrichomonas foetus. AB - Tritrichomonas foetus is a parasite of particular veterinary importance causing bovine tritrichomonosis, a sexually transmitted disease leading to infertility and abortion. The present review summarizes the current knowledge on potential mechanisms of pathogenicity of T. foetus, the immunology of host-parasite interaction in bovine tritrichomonosis, and the experimental model systems of this parasitic disease. PMID- 10816087 TI - Molecular epidemiology, pathogenesis, and genetics of the dimorphic fungus Histoplasma capsulatum. AB - Histoplasma capsulatum, the causative agent of the most common systemic fungal infection, histoplasmosis, has become subject to increasing study in parallel with rising prevalence of human immunodeficiency. This review presents a summary of the advances made in the investigation of H. capsulatum genomics, molecular epidemiology, pathogenesis, and molecular genetics. PMID- 10816088 TI - 'Infectious web'. PMID- 10816089 TI - An overview of bioactivation of chemical carcinogens. AB - Most environmental carcinogens require metabolic activation to reactive intermediates and are mutagenic in appropriate test systems. During the last decade, the cDNAs of numerous xenobiotic-metabolizing enzymes have been cloned. The individually expressed enzymes were used to study their substrate specificities and their inhibition by other compounds. Various enzymes were expressed directly in target cells of in vitro mutagenicity tests. This is illustrated in the present study for rat and human sulphotransferases (SULTs) expressed in Salmonella typhimurium TA1538. Numerous compounds were mutagenic in the new test system. Some of these promutagens were activated by several different SULT forms, whereas many other promutagens were activated with high selectivity by a specific enzyme form, but not by genetically closely related forms from the same species (e.g. allelic variants) or orthologous enzymes from other species. Similar findings have been made using recombinant test systems for specific forms of other classes of enzymes (e.g. cytochromes P450). This high selectivity in activation (and inactivation) may explain some organotropisms as well as species and inter-individual differences in the action of carcinogens. Many carcinogen-metabolizing enzymes are induced or inhibited by other xenobiotics. Such interactions can be exploited for chemo-prevention, which however may be carcinogen- and tissue-dependent. PMID- 10816090 TI - Modulation of signal-transduction pathways by chemopreventive agents. AB - For a disease such as cancer, where a number of alterations to normal cell function accumulate over time, there are several opportunities to inhibit, slow down or even reverse the process. Many of the changes which drive the disease process occur in cell-signalling pathways that regulate proliferation and apoptosis. As our knowledge of these complicated signalling networks improves, it is becoming clear that many molecules, both drugs and naturally occurring dietary constituents, can interact beneficially with deregulated pathways. Aspirin and other non-steroidal anti-inflammatory drugs, as well as natural compounds present in plants such as green vegetables and tea, can modulate signalling by affecting kinase activity and therefore phosphorylation of key molecules. Examples of pathways which can be modulated by these agents include activation of the transcription factor nuclear factor kappaB by tumour promoters or cytokines, signalling by growth factors through the growth-factor receptor/extracellular regulated protein kinase pathways and by a number of other molecules through the stress-activated c-Jun N-terminal kinase and p38 pathways. These mitogen activated protein kinase pathways regulate a number of transcription factors including c-Fos and c-Jun. Evidence exists, at least from in vitro experiments, that by targeting such pathways, certain dietary compounds may be able to restore abnormal rates of apoptosis and proliferation to more normal levels. PMID- 10816091 TI - Diet and colorectal cancer prevention. AB - The majority of cancers are sporadic and epidemiological estimates suggest that up to 80% of colorectal cancer is attributable to diet. Epidemiologically, cross sectional comparisons, case-control studies and trends in food intake show high rates of colorectal cancer in populations consuming diets high in meat and fat, and low in starch, NSP (non-starch polysaccharides, fibre) and vegetables. In general, prospective studies tend to support these findings although estimates of relative risk are not high. Existing prospective studies have however used crude indices of diet subject to substantial measurement error, and interactions with genetic polymorphisms in, for example, phase-I and -II enzymes have been studied only rarely. The association between meat consumption and colorectal cancer is usually attributed to the formation of heterocyclic amines in meat when it is cooked. In addition, in humans high-meat diets increase the level of nitrosatable material entering the colon so that faecal N-nitroso compounds (NOCs) increase in a dose-responsive manner following endogenous synthesis in the colon. Some of the mutations and guanine adducts accumulated during colorectal cancer progression are characteristic of alkylative damage, which would be compatible with NOC exposure. To date, NSP, resistant starch and vegetables have not reduced faecal NOC levels. PMID- 10816092 TI - Human metabolic pathways of dietary flavonoids and cinnamates. AB - Flavonoids and cinnamates are widespread phenolic secondary metabolites synthesized by plants for defensive purposes. Many foods and beverages contain high levels of phenolic compounds. Certain phenolics in the diet are particularly bioactive and have pronounced effects on mammalian cells. These effects, together with epidemiological studies and animal models, have led to the hypothesis that dietary phenolics contribute to the health benefits of a diet rich in fruit and vegetables. This paper examines the biochemistry of the uptake and metabolic route of two groups of plant phenolics, the flavonols and hydroxycinnamates. PMID- 10816093 TI - Cytotoxicity, genotoxicity and oxidative reactions in cell-culture models: modulatory effects of phytochemicals. AB - Much research effort has focused on the identification of phytochemicals in fruit and vegetables which exert beneficial effects. Our research examines modulatory effects of phytochemicals on cytotoxicity, genotoxicity and oxidative reactions in cell systems. Two examples of our studies are discussed. First, the potential beneficial effects of flavonoids are demonstrated. Flavonoids are reported to exhibit a wide variety of biological effects, including antioxidant and free radical-scavenging activities. The aim of the study was to determine if flavonoids could protect against H2O2-induced DNA damage, as measured by the comet assay, in Caco-2 and HepG2 cells. Both cell lines were supplemented with increasing concentrations of myricetin, quercetin and rutin for 24 h followed by exposure to H2O2 (50 microM) for 30 min. Exposure to H2O2 for 30 min at 37 degrees C resulted in significant DNA damage and pre-incubation with the flavonoids before H2O2 exposure significantly (P <0.05) protected Caco-2 and HepG2 cells against H2O2-induced DNA damage. Secondly, we illustrate the use of cellular models to study oxysterol-induced toxicity. Oxysterols are generated during the cooking and processing of foods and may be produced endogenously by the oxidation of membrane lipids. Recent findings suggest that oxysterols may modulate cytotoxicity by exerting effects on the induction of apoptosis. 7beta Hydroxycholesterol (7beta-OHC) and 25-hydroxycholesterol, both of which are commonly found in foods, were investigated for their abilities to induce apoptosis in a human monocytic blood cell line, U937, and in the human hepatoma cell line, HepG2 cells. U937 and HepG2 cells were incubated for up to 48 h with 30 microM oxysterol. 7beta-OHC induced apoptosis in U937 cells as measured by non random DNA fragmentation, condensed and fragmented nuclei, and the generation of hypodiploid cells. In contrast, oxysterols may induce cell death by a different mechanism in the hepatoma cells, possibly by necrosis. PMID- 10816094 TI - Chemical modulation of chemotherapy resistance in cultured oesophageal carcinoma cells. AB - Oesophageal carcinoma is a common form of cancer in developing countries, especially in the Caspian littoral and northern China. In contrast, it has a much lower incidence in Japan, the U.S.A. and western Europe. Certainly in the case of squamous cell oesophageal carcinoma, dietary composition, smoking, alcohol and exposure to nitrosamines are major risk factors that may partly explain the disease's geographical distribution. The prognosis for oesophageal carcinoma is generally poor, due to the high incidence of distant metastasis and local recurrence. Combination treatment with both cisplatin and 5-fluorouracil is the most common chemotherapy regime used. We have carried out a detailed study of sensitivity of two oesophageal cell lines: OC1 cells from a squamous carcinoma of a male patient, and OC2, a squamous carcinoma obtained from a female patient. Both cell lines are sensitive to Vinca alkaloids and doxorubicin, while being quite resistant to alkylating agents such as cisplatin and 1,3-bis-(2 chloroethyl)-1-nitrosourea. This pattern of resistance suggests a possible role for glutathione S-transferase (GST) and/or glutathione (GSH) in resistance, and would seem to exclude the multidrug resistance phenotype. Both cell lines possess mainly Pi-class GSTs, and have distinct levels of GSH, with OC2 possessing some 25% of the level of OC1 cells. Effects of a variety of modulating agents on the pattern of resistance, such as the GSH depleter, buthionine sulphoximine, and the GST inhibitor, ethacrynic acid, were determined. An unexpected observation was that ethacrynic acid appears to increase the level of GSH in both cell lines. PMID- 10816095 TI - The Nrf2 transcription factor contributes both to the basal expression of glutathione S-transferases in mouse liver and to their induction by the chemopreventive synthetic antioxidants, butylated hydroxyanisole and ethoxyquin. AB - An overview is provided of the cancer chemoprevention actions of phenolic antioxidants and 6-ethoxy-1,2-dihydro-2,2,4-trimethylquinoline (ethoxyquin). These agents principally appear to exert their beneficial effects through induction of phase II drug-metabolizing enzymes such as glutathione S-transferase (GST). The requirement for oxidative metabolism of the synthetic antioxidants to carbonyl-containing compounds, including quinones, in order that they can induce gene expression is discussed. Previous work has shown that the basic leucine zipper transcription factor Nrf2 is involved in induction of GST by the phenolic antioxidant butylated hydroxyanisole (BHA). Evidence is provided from a mouse possessing a targeted disruption of the Nrf2 gene that, in murine liver, the transcription factor regulates basal expression of several class Alpha and class Mu GST subunits, but not class Pi GST. In the Nrf2 knock-out mouse, hepatic induction of class Alpha and class Mu GST by BHA and the synthetic antioxidant ethoxyquin is similarly impaired, suggesting that these agents affect gene activation by a related mechanism. Significantly, residual induction of GST by antioxidants is apparent in the Nrf2 mutant mouse, indicating the existence of an alternative mechanism of gene activation. PMID- 10816096 TI - Cytochrome P450s and chemoprevention. AB - The cytochrome P450 mono-oxygenase system represents a major defence against chemical challenge from the environment, constituting part of an adaptive response mounted by an organism following exposure to harmful agents. Cytochrome P450s are also able to catalyse the activation of compounds to toxic products, and participate in a variety of essential 'housekeeping' functions, such as biosynthesis of steroid hormones and fatty acid oxidation. It is clear that the modulation of expression of these enzymes can have a significant effect on chemical toxicity, carcinogenicity and mutagenicity. The concept of cancer chemoprevention, i.e. the administration of a (non-toxic) chemical or dietary component in order to prevent neoplastic disease or to inhibit its progression, is an attractive one. Despite this, relatively little work has been done to characterize the ability of putative chemopreventive agents to modulate P450 expression, or to understand the interaction between P450s and chemopreventive agents. Before chemopreventive treatment can become a reality, it is essential that this complex issue is addressed; for instance, it is likely that any single chemopreventive agent will induce more than one P450 isoenzyme, and while altered expression of a particular P450 may attenuate the effects of one toxic agent, the effects of others might well be potentiated. Our laboratory has created a transgenic mouse line in which the rat CYP1A1 promoter drives expression of the beta-galactosidase gene. These mice can be used to define which compounds act via the Ah receptor, in which tissues, and at which stage of development. We are currently developing another mouse line in which beta1-galactosidase expression is controlled by the mouse GstA1 promoter, allowing us to define the role of the antioxidant responsive element in the action of chemopreventive agents. Finally, using cre-loxP transgenic technology, we have generated a mouse line in which P450 reductase can be deleted in a conditional, i.e. tissue-specific, manner, permitting us to investigate the role of P450s in chemoprevention in a more defined manner. PMID- 10816097 TI - Regulation of survival signals from the insulin-like growth factor-I receptor. AB - Suppression of apoptosis by survival factors is important for the maintenance of normal tissue homoeostasis and the response to infection or injury. Survival factors such as insulin-like growth factor-I (IGF-I) initiate a signalling cascade that starts by tyrosine phosphorylation of substrates leading to the activation of serine kinases that modulate the activity of members of the Bcl-2 family, which regulates the apoptotic machinery in most cells. Tumour cells often have enhanced survival mechanisms due either to up-regulation of the IGF-I receptor and its ligands or to loss of function of a phosphatase (PTEN) that regulates part of this survival pathway. The C-terminus of the IGF-I receptor appears to be a regulatory domain for the anti-apoptotic activity of this receptor, and certain residues within the C-terminus are essential for this regulatory activity. Knowledge of the proteins and pathways, which interact with these C-terminal domains, should lead us to ways of modulating IGF-I-mediated survival in tumours. PMID- 10816098 TI - Role of the BH3 (Bcl-2 homology 3) domain in the regulation of apoptosis and Bcl 2-related proteins. AB - The Bcl-2 family of proteins play a prominent role in the regulation of apoptosis. From the initial identification of bcl-2 as an oncogene in follicular lymphoma through genetic studies in Caenorhabditis elegans to recent functional studies focusing on the importance of mitochondrial events in cell death signalling, the members of this protein family continue to be implicated in pivotal decision points regarding the survival of the cell. The family can be divided into two classes: those such as Bcl-2 and Bcl-xL that suppress cell death, and others, such as Bak and Bax, that appear to promote apoptosis. The Bcl 2 family is characterized by specific regions of homology termed Bcl-2 homology (BH1, BH2, BH3, BH4) domains, which are critical to the function of these proteins, including their impact on cell survival and their ability to interact with other family members and regulatory proteins. The identification of the BH3 domain as a potent mediator of cell death has led to the emergence of an additional family of proapoptotic proteins (such as Bad, Bik, Bid and Hrk) that share identity with Bcl-2 only within this death domain. These BH3-only proteins may be part of a regulatory network serving to integrate cell survival and death signals, an assertion that is supported by the identification of a BH3-only protein, Egl-1, as part of the central core of cell death signalling in C. elegans. While the mechanism of action of the BH3-only proteins remains unclear, recent studies on the regulation of critical protein-protein interactions and activity of Bad by phosphorylation in response to growth factor signalling suggest that the active state of BH3-only proteins may be regulated by post translational modification. Additional modes of regulation, such as transcriptional, translational and subcellular localization, are also likely to be important. PMID- 10816099 TI - GSH extrusion and and the mitochondrial pathway of apoptotic signalling. AB - New evidence suggests that physiological and damaging agents activate two different pathways of apoptotic signalling, which are mediated by protein-protein interactions and mitochondrial alterations respectively. The two pathways converge at the activation of caspase 3, the key effector of the execution phase of apoptosis, thus giving similar final results. The knowledge that different biochemical routes exist allows us to re-evaluate previous apparently contradictory results concerning the events occurring during apoptosis, and their respective roles. In particular, this applies to the role of oxidative stress and redox imbalance in the signal transduction events of apoptosis. It now appears that oxidative alterations are absent, or at least unnecessary, for the development of the physiological pathway. Instead, clear indications are emerging showing that redox imbalance is required for the damage-induced mitochondrial pathway. This is suggested by the finding that the depletion of glutathione, a common event in damage-induced apoptosis, is necessary and sufficient to induce cytochrome c release, the key event of this pathway. A model is proposed with GSH efflux as the backbone of the damage-induced apoptotic pathway. PMID- 10816100 TI - Electrochemical investigation of cellobiose oxidation by cellobiose dehydrogenase in the presence of cytochrome c as mediator. AB - An important aspect of the cytochrome c electrochemistry is the possibility of coupling the 'heterogeneous reactions' with other redox enzymes. Cellobiose dehydrogenase, a 89170 Da glycoprotein that contains both FAD and a b-type haem as prosthetic groups, donates electrons to a number of acceptors, including cytochrome c. While haem b is surrounded mainly by acidic amino acids, cytochrome c displays positive charged lysine groups around the haem site. Thus a fast reaction between both proteins is explicable. In the presence of cellobiose, a catalytic current was observed, owing to the interaction of cellobiose dehydrogenase with electrostatically adsorbed cytochrome c. Adsorption of cytochrome c provides a technological model surface for vectorial electron transfer. PMID- 10816101 TI - Two-photon fluorescence excitation in detection of biomolecules. AB - Two-photon fluorescence excitation has been found to be a very powerful method for enhancing the sensitivity and resolution in far-field light microscopy. Two photon fluorescence excitation also provides a substantially background-free detection on the single-molecule level. It allows direct monitoring of formation of labelled biomolecule complexes in solution. Two-photon excitation is created when, by focusing an intensive light source, the density of photons per unit volume and per unit time becomes high enough for two photons to be absorbed into the same chromophore. In this case, the absorbed energy is the sum of the energies of the two photons. In two-photon excitation, dye molecules are excited only when both photons are absorbed simultaneously. The probability of absorption of two photons is equal to the product of probability distributions of absorption of the single photons. The emission of two photons is thus a quadratic process with respect to illumination intensity. Thus in two-photon excitation, only the fluorescence that is formed in the clearly restricted three-dimensional vicinity of the focal point is excited. We have developed an assay concept that is able to distinguish optically between the signal emitted from a microparticle in the focal point of the laser beam, and the signal emitted from the surrounding free labelled reagent. Moreover, the free labels outside the focal volume do not contribute any significant signal. This means that the assay is separation-free. The method based on two-photon fluorescence excitation makes possible fast single step and separation-free immunoassays, for example, for whole blood samples. Since the method allows a separation-free assay in very small volumes, the method is very useful for high-throughput screening assays. Consequently we believe that two-photon fluorescence excitation will make a remarkable impact as a research tool and a routine method in many fields of analysis. PMID- 10816102 TI - Phosphorescent porphyrin probes in biosensors and sensitive bioassays. AB - Platinum(II) and palladium(II) complexes of porphyrins and related tetrapyrrolic pigments emit strong phosphorescence at room temperatures, which is characterized by long lifetimes falling into the sub-millisecond range and long-wave spectral characteristics. These features make the dyes useful as probes for a number of bioanalytical applications, particularly those employing time-resolved fluorescent detection. They can provide high sensitivity and selectivity, together with rather simple instrumental set-up. A number of analytical systems are now under development that are based on the use of phosphorescent porphyrin probes. Experimental results are presented on the following systems: (i) fibre optic phosphorescence lifetime-based oxygen sensor on the basis of hydrophobic platinum-porphyrins and development of advanced sensing materials and prototype instrumentation; (ii) practical applications of the optical oxygen sensor, including a sensitive immunosensor that employs glucose oxidase labels, a rapid screening method for cell viability in microtitre-plate format, non-destructive measurement of oxygen in packaged foods and reagentless biosensors for metabolites (glucose, lactate); and (iii) the use of water-soluble platinum- and palladium-porphyrins as labels for ultra-sensitive time-resolved phosphorescence immunoassays. PMID- 10816103 TI - Double-stranded DNA liquid-crystalline dispersions as biosensing units. AB - Three different approaches to constructing biosensing units based on double stranded (ds) DNA molecules, capable of detecting various biologically active compounds, are considered. The first approach is based on the abnormal optical activity of the liquid-crystalline dispersion formed from ds DNA molecules, modified by relevant physical factors or treated with biologically active compounds. The second one is based on the abnormal optical activity of the liquid crystalline dispersions formed first from the ds DNA and then treated with coloured biologically active compounds. The third one is based on the abnormal optical activity, specific to particles of the liquid-crystalline dispersions, where the neighbouring DNA molecules are crosslinked by artificial polymeric bridges. These approaches permit the detection of biologically relevant compounds of various origins. PMID- 10816104 TI - Rapid antibody biosensor assays for environmental analysis. AB - Traditionally, biosensor development has focused on molecules with a defined metabolic role that can be exploited by enzyme-based systems. Antibodies have the ability to move beyond this range of analytes, and are particularly useful in detecting small, hapten molecules. Electrochemically based biosensor developments have been less fruitful in this regard, as enzyme labelling is required, and such assays require the separation from bound and unbound species. These separations and the removal of background signals result in the increased complexity of the assay format, making it unsuitable for rapid sensor analysis. We have developed an electrochemical sensor based on antibodies that does not require the separation of bound and unbound molecules in a competition immunoassay format. This removes the need for several washing and separation steps as is normally employed in this type of assay. This allows single-step immunoassays to be performed using this system, and also allows for the real-time monitoring of antibody-antigen interactions. We have shown that such assays are possible in both batch and flow-injection formats and we are currently developing an assay for the pesticide atrazine. Tentative results show that analysis with this system is possible in the p.p.m. to p.p.b. range. PMID- 10816105 TI - Direct electron transfer catalysed by enzymes: application for biosensor development. AB - The ability to catalyse an electrode reaction via direct (mediatorless) electron transfer has been demonstrated for a number of redox enzymes. In the case of mediatorless electron transfer, the electron is transferred directly from the electrode to the substrate molecule via the active site of the enzyme, or vice versa. The electron itself is the second substrate for the reaction. An important point characterizing bioelectrocatalysis is the catalytic removal of the reaction over-voltage. Therefore the enzyme attached to the electrode is able to catalyse electrode reaction and forms a 'molecular transducer'. The substrate can be detected by potentiometric measurement of the removal of reaction over-voltage. The enzyme laccase is able to catalyse the reaction of oxygen electroreduction. Therefore a laccase molecular layer attached to the electrode surface forms an oxygen transducer. The formation of the layer results in a change of the electrocatalytic feature of the electrode. Laccase label coupled with either ligand or receptor allows the detection of ligand-receptor complex formation/dissociation on the electrode surface. The detection is virtually reagentless. The substrates for the reaction are molecular oxygen and the electron itself. Numerous reagentless immunosensors of different formats (competitive, displacement and sandwich) have been developed, as well as the reagentless detection system for immunofiltration/immunochromatography. PMID- 10816106 TI - The use of electrochemically grown polymers on metallized electrodes to reduce electrode fouling in biological matrices. AB - The use of electrochemically grown polymers has expanded dramatically in the last couple of years, and they are now well established as membranes for immobilizing components. The evidence here for their anti-fouling properties is good. The poly(1,3-diaminobenzene)-covered electrodes performed well in the buffer, urine, plasma and serum samples, but not so well in the blood. The Ru/Rh/Pt, Rh/Rh and the Pt-on-glassy carbon electrodes covered with poly(1,3-diaminobenzene) were the best electrodes in the blood. The Pt disc seemed to exhibit the largest irrepeatability in most of the biological matrices. PMID- 10816107 TI - The multifunctional protein AtMFP2 is co-ordinately expressed with other genes of fatty acid beta-oxidation during seed germination in Arabidopsis thaliana (L.) Heynh. AB - In germinating oilseeds peroxisomal fatty acid beta-oxidation is responsible for the mobilization of storage lipids. This pathway also occurs in other tissues where it has a variety of additional physiological functions. The central enzymatic steps of peroxisomal beta-oxidation are performed by acyl-CoA oxidase (ACOX), the multifunctional protein (MFP) and 3-ketoacyl-CoA thiolase (thiolase). In order to investigate the function and regulation of beta-oxidation in plants it is first necessary to identify and characterize genes encoding the relevant enzymes in a single model species. Recently we and others have reported on the cloning and characterization of genes encoding four ACOXs and a thiolase from the oilseed Arabidopsis thaliana. Here we identify a gene encoding an Arabidopsis MFP (AtMFP2) that is induced transiently during germination. The pattern of AtMFP2 expression closely reflects changes in the activities of 2-trans-enoyl-CoA hydratase and L-3-hydroxyacyl-CoA dehydrogenase. Similar patterns of expression have previously been reported for ACOX and thiolase genes. We conclude that genes encoding the three main proteins responsible for beta-oxidation are co-ordinately expressed during oilseed germination and may share a common mechanism of regulation. PMID- 10816108 TI - Studying metabolic regulation in human muscle. AB - Phosphorus magnetic resonance spectroscopy, despite some limitations, is a valuable non-invasive window on muscle metabolism in vivo, particularly oxidative ATP synthesis. A number of experiments have shown this to be dominated by closed loop feedback mechanisms: a well-known model posits regulation by ADP, but there are others, difficult to distinguish experimentally. Moreover the contribution of open-loop control mechanisms ('feed forward' or 'parallel activation') in vivo remains controversial. Progress will require more precise data, better integrated with other measurements (e.g. muscle oxygenation), and improvement of the conceptual tools appropriate to such studies, where data are limited and steady state assumptions frequently inapplicable. PMID- 10816109 TI - Use of in vivo and in vitro techniques for the study of the effects of insulin on hepatic triacylglycerol secretion in different insulinaemic states. AB - This review illustrates how the use of several in vitro and in vivo techniques was necessary to show that the effect of insulin on hepatic triacylglycerol (TAG) secretion in the rat depends on the prior physiological state of the animal. The effect of insulin was always inhibitory when cultured cells were used, irrespective of the physiological state of the donor rats. By contrast, when perfused livers were used, insulin stimulated TAG secretion by livers isolated from fed, normoinsulinaemic rats, but inhibited it in livers from fasted or streptozotocin diabetic animals. This switch in insulin action was also shown to occur in vivo in experiments that involved the liver-specific targeting of both insulin (delivered within liposomes) and labelled fatty acids (delivered as cholesteryl esters within very-low-density lipoprotein remnants) in awake, unrestrained rats during a euglycaemic clamp. It is concluded that observations obtained with perfused liver preparations are more representative of the actual changes that occur in vivo with respect to the effects of insulin on hepatic TAG secretion. PMID- 10816110 TI - The use of in situ haemoglobin-free perfused liver in metabolic-control analysis. AB - In this study the network of ATP-consuming and -producing reactions, interacting via the cytosolic ATP/ADP+P(i) system, was studied for the first time in an intact organ, the isolated perfused rat liver, using top-down metabolic-control analysis. Flux control in the metabolically resting state (only oxidative phosphorylation and ion pumps active) was only in the ATP-consumers, whereas in the metabolically active state (biosyntheses and ion-pumping ATPases active) most control over oxidative phosphorylation was in itself and some control was in ion pumping ATPases. All ATP-consumers had high control over themselves, and ion pumping ATPases had high negative control over the other ATP-consuming branches. Oxidative phosphorylation had high positive control over ATP-consuming reactions except for ion-pumping ATPases. It is concluded that in the active state ATP consumers compete for cytosolic ATP, but as the ion-pumping ATPases and oxidative phosphorylation are less sensitive towards the cytosolic ATP, other ATP-consumers have low control over these pathways. PMID- 10816111 TI - Perfused heart studies to investigate lipid metabolism. AB - The isolated perfused heart preparation is an invaluable model for investigating metabolism in a variety of physiological and pathological states. It avoids confounding systemic factors (e.g. endocrine, metabolic and work load changes) and permits simultaneous measurement of mechanical function. The ability to measure arteriovenous concentration differences across the myocardium and the coronary flow rate, together with the use of radiolabelled substrates, permits assessment of substrate assimilation and disposition of most potential energetic substrates. In the case of lipids, metabolism of non-esterified fatty acids has been extensively investigated in the perfused rat heart, but fatty acids may also be derived from circulating triacylglycerols (TAG) in lipoproteins [chylomicrons, very-low-density-lipoprotein (VLDL)]. TAG requires initial hydrolysis by the endothelial enzyme lipoprotein lipase and hence an intact heart preparation is vital to maintain tissue structural integrity. Chylomicron-TAG utilization and fate (oxidation, tissue-lipid deposition) in isolated working hearts has been studied using chylomicrons obtained from thoracic-duct catheters. However, lack of availability of sufficient quantities of VLDL has hindered examination of their cardiac utilization; the recent development of a technique to produce large quantities of radio-labelled rat VLDL has facilitated these studies and established that VLDL-TAG is an important metabolic substrate for working heart. Results relating to myocardial utilization of VLDL-TAG under varying physiological (lactation) and pathological (endotoxinaemia) conditions will be presented. The putative role of VLDL as a regulator of cardiac lipid metabolism will also be discussed. PMID- 10816112 TI - NMR spectroscopy and imaging of the neonatal brain. AB - Magnetic resonance spectroscopy and imaging provide unique information about the brain to the biochemist and the clinician. In particular, the ability to image metabolites other than water and to get detailed information about dynamic cellular processes (such as blood flow, blood oxygenation and cell swelling) is leading to many new insights into brain function and dysfunction. This review describes the use of old and new NMR techniques which demonstrate that mitochondrial dysfunction plays an important role in the cell death that occurs following an hypoxic-ischaemic insult to the neonatal brain. PMID- 10816113 TI - Adipocyte studies: systems for investigating effects of growth hormone and other chronically acting hormones. AB - Adipose tissue is very amenable to study in vitro. Collagenase digestion yields free adipocytes which usually respond well to acute stimulation/inhibition by hormones and other factors. Chronic effects of hormones are best studied using explants of adipose tissue which, from some species (e.g. sheep), can be maintained in culture for up to a week without loss of function. Alternatively, preadipocytes can be readily isolated from adipose tissue and induced to proliferate and differentiate in culture, while various adipocyte-like cell-lines have been established, which can be used for chronic studies. Use of these various systems for investigating the mechanisms of action of growth hormone are described. PMID- 10816114 TI - The isolated hepatocyte preparation: 30 years on. AB - A method for the preparation of intact rat hepatocytes in high yield was first described in 1969. The procedure involved digestion of hepatic tissue by perfusion of the liver with crude collagenase; later, purified collagenase without other enzymic additions was shown to be ineffective. Recently it has been discovered that the combination of purified collagenase plus elastase is superior to crude collagenase in that it consistently provides high yields of undamaged hepatocytes. The isolated hepatocyte preparation has proved particularly useful for the study of mechanisms responsible for long-range interactions within the cell. These can be studied over prolonged time courses and in the presence of graded concentrations of specific inhibitors. Studies of this kind have demonstrated a close relationship between cytoplasmic metabolic flows and mitochondrial forces and have also revealed that the cytoplasmic and mitochondrial free NAD-linked redox potentials are maintained by energy-dependent reactions. PMID- 10816115 TI - Hepatocyte couplets. AB - Hepatocyte couplets, by retaining functional and morphological polarity similar to that of hepatocytes in situ, are a valuable in vitro model to study mechanisms of bile secretion, cholestasis and hepatocellular injury. They have been useful in studies of the hormonal control of bile formation and are suitable for morphological studies. The availability of periportal- and perivenous-enriched couplet populations now allows a zonal perspective. Their contribution to our understanding of regulatory aspects of hepatobiliary dysfunction due to toxicological or cholestatic insult, as well as its reversion by using hepatoprotective agents, is still at an early stage. The next few years should see further exiting contributions to our understanding of hepatobiliary function and dysfunction. PMID- 10816116 TI - Lipid metabolism in peroxisomes: enzymology, functions and dysfunctions of the fatty acid alpha- and beta-oxidation systems in humans. AB - Peroxisomes are subcellular organelles present in virtually all eukaryotic cells catalysing a number of indispensable functions in cellular metabolism. The importance of peroxisomes in man is stressed by the existence of an expanding group of genetic diseases in which there is an impairment in one or more peroxisomal functions. One of the major functions of peroxisomes concerns their role in lipid metabolism, which includes: (i) fatty acid betaoxidation; (ii) ether phospholipid synthesis; (iii) fatty acid alpha-oxidation; and (iv) isoprenoid biosynthesis. In this paper, we review the current state of knowledge concerning the peroxisomal fatty acid alpha- and beta-oxidation systems with particular emphasis on the enzymes involved and the various disorders of fatty acid oxidation in peroxisomes. We also pay attention to the fact that some of the metabolites that accumulate as the result of a defect in peroxisomal alpha- and/or beta-oxidation are activators of members of the family of nuclear receptors, including peroxisome-proliferator-activated receptor alpha. PMID- 10816117 TI - The molecular basis of lysosomal storage diseases and their treatment. AB - The lysosomal system is the main intracellular mechanism for the catabolism of naturally occurring endogenous and exogenous macromolecules and the subsequent recycling of their constituent monomeric components. It also plays an important part in processing essential metabolites. A genetic defect in a protein responsible for maintaining the lysosomal system results in the accumulation within lysosomes of partially degraded molecules, the initial step in the process leading to a lysosomal storage disease. The defective protein can be a luminal lysosomal enzyme or protein cofactor, a lysosomal membrane protein or a protein involved in the post-translational modification or transport of lysosomal proteins. Over 40 lysosomal storage diseases are known and they have a collective incidence of approximately 1 in 7000-8000 live births. Most of the genes for the lysosomal proteins have been cloned, permitting mutation analysis in individual cases. This information can be used for genotype/phenotype correlation, genetic counselling and the selection of patients for novel forms of therapy, such as substrate deprivation or dispersal, enzyme replacement, bone-marrow transplantation and gene transfer. PMID- 10816118 TI - Does the mitochondrial transcription-termination complex play an essential role in controlling differential transcription of the mitochondrial DNA? AB - The mechanism of mitochondrial transcription is well documented although the method of regulation remains obscure. The mitochondrial transcription-termination complex, mTERF, holds a key position in determining the fate of heavy-strand promotor-initiated transcripts and has been suggested as a candidate in the regulation of mitochondrial DNA (mtDNA) transcription. We report here the first example of a modulation of mTERF-complex binding activity concomitant with a differential mtDNA transcription rate. We suggest that these observations are indicative of a method of intra-organellar transcriptional fine tuning. PMID- 10816119 TI - Muscle fibres: applications for the study of the metabolic consequences of enzyme deficiencies in skeletal muscle. AB - Mitochondrial function in saponin-permeabilized muscle fibres can be studied by high-resolution respirometry, laser-excited fluorescence spectroscopy and fluorescence microscopy. We applied these techniques to study metabolic effects of changes in the pattern of mitochondrial enzymes in skeletal muscle of patients with chronic progressive external ophthalmoplegia or Kearns-Sayre syndrome harbouring large-scale deletions of mitchondrial DNA (mtDNA). In all patients combined deficiencies of respiratory chain enzymes containing mitochondrially encoded subunits were observed. The citrate synthase-normalized activity ratios of these enzymes decreased linearly with increasing mtDNA heteroplasmy. This indicates the absence of any well-defined mutation thresholds for mitochondrial enzyme activities in the entire skeletal muscle. We applied metabolic control analysis to perform a quantitative estimation of the metabolic influence of the observed enzyme deficiencies. For patients with degrees of mtDNA heteroplasmy below about 60% we observed at almost normal maximal rates of respiration an increase in flux control coefficients of complexes I and IV. Permeabilized skeletal-muscle fibres of patients with higher degrees of mtDNA heteroplasmy and severe enzyme deficiencies exhibited additionally decreased maximal rates of respiration. This finding indicates the presence of a 'metabolic threshold' which can be assessed by functional studies of muscle fibres providing the link to the phenotypic expression of the mtDNA mutation in skeletal muscle. PMID- 10816120 TI - Function of the mitochondrial outer membrane as a diffusion barrier in health and diseases. AB - The mitochondrial outer membrane separates the intermembrane space from the cytosol. The whole exchange of metabolites, cations and information between mitochondria and the cell occurs through the outer membrane. Experimental evidence is reviewed supporting the hypothesis of dynamic ADP compartmentation within the intermembrane space. The outer membrane creates a diffusion barrier for small molecules (adenine nucleotides, creatine phosphate, creatine etc.) causing rate-dependent concentration gradients as a prerequisite for the action of ADP shuttles via creatine kinases or adenylate kinases. If the outer membrane becomes leaky, cytochrome c and apoptosis-inducing factor can be released, leading to apoptosis, and as a bioenergetic consequence the cytosolic phosphorylation potential decreases. Leaky outer membranes can be detected in saponin-skinned fibres with spectrophotometric and oxygraphic methods. This is of special interest in respect to acute impairment of mitochondria during ischaemia/reperfusion. PMID- 10816121 TI - Mitochondria and cell death. AB - Mitochondria play a central role in both apoptosis and necrosis through the opening of the mitochondrial permeability transition pore (MPTP). This is thought to be formed through a Ca(2+)-triggered conformational change of the adenine nucleotide translocase (ANT) bound to matrix cyclophilin-D and we have now demonstrated this directly by reconstitution of the pure components. Opening of the MPTP causes swelling and uncoupling of mitochondria which, unrestrained, leads to necrosis. In ischaemia/reperfusion injury of the heart we have shown MPTP opening directly. Recovery of hearts correlates with subsequent closure, and agents that prevent opening or enhance closure protect from injury. Transient MPTP opening may also be involved in apoptosis by initially causing swelling and rupture of the outer membrane to release cytochrome c (cyt c), which then activates the caspase cascade and sets apoptosis in motion. Subsequent MPTP closure allows ATP levels to be maintained, ensuring that cell death remains apoptotic rather than necrotic. Apoptosis in the hippocampus that occurs after a hypoglycaemic or ischaemic insult is triggered by this means. Other apoptotic stimuli such as cytokines or removal of growth factors also involve mitochondrial cyt c release, but here there is controversy over whether the MPTP is involved. In many cases cyt c release is seen without any mitochondrial depolarization, suggesting that the MPTP does not open. Recent data of our own and others have revealed a specific outer-membrane cyt c-release pathway involving porin that does not release other intermembrane proteins such as adenylate kinase. This is opened by pro-apoptotic members of the Bcl-2 family such as BAX and prevented by anti-apoptotic members such as Bcl-X(L). Our own data suggest that this pathway may interact directly with the ANT in the inner membrane at contact sites. PMID- 10816122 TI - The mitochondrial trifunctional protein: centre of a beta-oxidation metabolon? AB - The trifunctional enzyme comprises three consecutive steps in the mitochondrial beta-oxidation of long-chain acyl-CoA esters: 2-enoyl-CoA hydratase, 3 hydroxyacyl-CoA dehydrogenase and 3-ketoacyl-CoA thiolase. Deficiencies in either 3-hydroxyacyl-CoA dehydrogenase activity, or all three activities, are important causes of human disease. The dehydrogenase and thiolase have a requirement for NAD+ and CoA respectively, whose levels are conserved within the mitochondrion and thus provide possible means for control and regulation of beta-oxidation. Using analysis of the intact CoA ester intermediates produced by the complex, we have examined the sensitivity of the complex to NAD+/NADH and acetyl-CoA. We consider the evidence for channelling within the trifunctional protein and propose a model for a beta-oxidation 'metabolon'. PMID- 10816123 TI - The carnitine acyltransferases: modulators of acyl-CoA-dependent reactions. AB - Carnitine and carnitine acyltransferases were thought to be merely a mechanism for the rapid transfer of activated long-chain fatty acids into the mitochondrion for beta-oxidation, until enzymologists came along. By kinetic, physical and localization studies, eight different mammalian carnitine acyltransferases have been characterized. Of these, five have been cloned and sequenced. The carnitine :acylcarnitine exchange carrier, first characterized in mitochondria, has now been demonstrated immunologically in peroxisomal membranes too. This cell-wide carnitine system consisting of at least six proteins linking at least four intracellular pools of acyl-CoA that supply a multitude of lipid metabolic pathways is clearly more complex than was first thought. In this article, I describe the location and properties of the components to show how they can modulate acyl-CoA-dependent reactions in the cell. PMID- 10816124 TI - Leishmania pyruvate kinase: the crystal structure reveals the structural basis of its unique regulatory properties. AB - Glycolysis occupies a central role in cellular metabolism, and is of particular importance for the catabolic production of ATP in protozoan parasites such as Leishmania and Trypanosoma. In these organisms pyruvate kinase plays a key regulatory role, and is unique in responding to fructose 2,6-bisphosphate as allosteric activator. The determination of the crystal structure of the first eukaryotic pyruvate kinase in the T-state (the inactive or 'tense' conformation of allosteric enzymes) is described. A comparison of the effector sites of the Leishmania and yeast enzymes reveals the structural basis for the different effector specificity. Two loops, comprising residues 443-453 and 480-489, adopt very different conformations in the two enzymes, and Lys-453 and His-480 that are a feature of trypanosomatid enzymes provide probable ligands for the 2-phospho group of the effector molecule. These and other differences offer an opportunity for the design of drugs that would exploit regulatory differences between parasite and host. PMID- 10816125 TI - Leucocyte populations and cytokine regulation in human uteroplacental tissues. AB - Human endometrial tissue and the decidualized endometrium in pregnancy contain relatively large numbers of leucocytes, the proportions of which vary during both the menstrual cycle and early pregnancy. CD3+ T-cells, CD14+ macrophages and a population of phenotypically unusual CD3-CD16-CD56++ large granular lymphocytes (LGL) are present, whereas B-cells are virtually absent. Relative T-cell numbers decrease in first-trimester decidua; T-cells are therefore unlikely to have an important role in the immunological maintenance of pregnancy but could be more important in implantation, when their relative numbers are greater. Extensive numbers of class II MHC-positive tissue macrophages in both the endometrium and placenta will provide an immediate antigen non-specific host defence to infection at this important site. Nevertheless, most attention has focused on a role for the predominant LGL endometrial cell population in the implantation and maintenance of pregnancy because, at the time of implantation, these LGLs comprise 70-80% of all leucocytes in the endometrium. It is now well recognized that there is substantial and complex cytokine activity within human uteroplacental tissues; both leucocytic and non-leucocytic cells have been shown to be capable of producing a significant array of cytokines. However, to avoid excessive pathological sequelae, such cytokine activity must be locally regulated. This has been highlighted by recent reports indicating that abnormal Th1-type cytokine responses could be a reason for immunological reproductive failure in women. Key cytokines controlling differentiation into a Thl (interleukin 12) or Th2 (interleukin 4) type pattern both exist in unusual molecular forms at the human maternal-fetal tissue interface and hence might be fundamental regulatory elements controlling cytokine action locally by an antagonistic action. PMID- 10816126 TI - Decidual natural-killer-cell interaction with trophoblast: cytolysis or cytokine production? AB - At the implantation site, the uterine mucosa (decidua) is infiltrated by large numbers of natural killer (NK) cells. These NK cells are in close contact with the invading fetal trophoblast and we have proposed that they might be the effector cells that control the implantation of the allogeneic placenta. Recent characterization of NK cell receptors and their HLA class I ligands has suggested potential mechanisms by which NK cells might interact with trophoblast. However, what happens as a result of this interaction is not clear. The traditional method for investigating NK cell function in vitro is the protection from lysis of target cells by expression of HLA class I antigens. This might not be an accurate reflection of what happens in vivo. Another function of NK cells is the production of cytokines on contact with target cells. This could be an important outcome of the interaction between decidual NK cells and trophoblast. Decidual NK cells are known to produce a variety of cytokines; trophoblast cells express receptors for many of these cytokines, indicating that they can potentially respond. In this way, decidual NK cells have a significant influence on trophoblast behaviour during implantation. PMID- 10816127 TI - Growth factor-extracellular matrix synergy in the control of trophoblast invasion. AB - At the periphery of the human placenta, trophoblast attaches to the uterine wall. The tissue interface contains many anchoring sites, with cytotrophoblast columns that form bridges between the overlying extraembryonic (villous) mesenchyme and the maternal decidual stroma beneath. From the periphery of these columns, large numbers of trophoblast cells detach, migrate through the decidua and eventually colonize and transform maternal arteries. In this way the placenta increases and gives priority to the maternal blood supply to the conceptus. We have shown that when early villous tissue is explanted on a collagen gel in serum-free medium, anchoring-site morphogenesis occurs. Thus, in the presence of placental mesenchyme but in the absence of maternal cells, contact with a permissive extracellular matrix (ECM) is necessary and sufficient for cytotrophoblast column development. Proliferation of trophoblast occurs, followed by differentiation into a columnar cell phenotype in which cells remain attached to one another and to the ECM. At this stage, interaction between fibronectin and integrin alpha5beta1 at the cell surface stabilizes the column and the cells remain as a contiguous multilayered sheet. However, the addition of serum-free conditioned medium from first-trimester placental fibroblasts stimulates cytotrophoblast to detach from the distal column and migrate in streams across the ECM. The removal of insulin-like growth factor I (IGF-I) from the fibroblast medium decreases streaming activity, whereas the addition of exogenous IGF-I (10 ng/ml) to serum free medium produces a streaming phenotype. In contrast, transforming growth factor beta1 (10 ng/ml) maintains the cells in a tight sheet. These results suggest the possibility of a paracrine interaction between villous mesenchyme and cytotrophoblast in anchoring sites to stimulate the infiltration of the maternal ECM by trophoblast. Such a mechanism would be self-limiting because the signal diminishes with distance from the placenta. PMID- 10816128 TI - Structure-function relations in the human placenta. AB - The human haemochorial placenta is a complex and dynamic interface between embryonic and maternal tissues. A myriad array of compounds has been identified at this interface, some of which exert local effects which might be important in maintaining the integrity of the organ. These compounds are diverse in nature and function; they include enzymes, hormones and bioactive peptides. Successful nidation requires the synchronization of endometrial maturation and embryonic development. The complex nature of this interface requires the application of sound sampling strategies. The new stereological methods have thrown fresh light on the growth and development of the human placenta. These methods permit the objective, quantitative description of morphology by efficient design-based methods. This approach has permitted a better definition of the functional morphology of the placenta. Applications of these methodologies are providing a spatial and temporal framework on which to lay the new physiological and molecular information. Here we review the essential features of the stereological approach, identify useful structural quantities and provide some examples of their application. The problems associated with the quantification of immunocytochemistry are illustrated with the use of immunoreactivity to insulin like growth factor I receptor in normal placentae and in pre-eclampsia. Although stereology can provide useful quantitative information about the structure of this dynamic tissue, other anatomical methods that could be applied to better define the relationships between structure and function will be discussed. These include confocal microscopy, to examine the dynamic physiological interactions of the different tissue compartments, and low-temperature electron microscopy techniques such as cryosubstitution, to allow better access to the biochemical information resident in the tissue. The complex and dynamic nature of the tissue requires a multidisciplinary approach; central to these investigations is a comprehensive understanding of its fine structure. PMID- 10816129 TI - HLA-G in the human placenta: expression and potential functions. AB - HLA-G is a non-classical class I molecule specifically expressed in the placenta, suggesting that it might have a physiological function at the materno-foetal interface. The structural characteristics of HLA-G, the placental pattern of expression and the functional properties of this class Ib glycoprotein in vitro are described and evaluated in the context of pregnancy. The possible anti-viral function of HLA-G, its modulatory role of natural killer cell activity and its likely non-immunological functions are discussed. PMID- 10816130 TI - Role of hormone-controlled T-cell cytokines in the maintenance of pregnancy. AB - Human CD4 T helper lymphocytes can be subdivided into at least three distinct functional subsets on the basis of their cytokine secretion profiles. One type of CD4+ lymphocyte, T helper 1 (Th1), produces interferon (IFN)-gamma and tumour necrosis factor beta, a second type (Th2) produces interleukin (IL)-4 and IL-5 and a third type (Th0) produces both Th1 and Th2 cytokines. The apparent paradox that embryos are not rejected by the maternal immune system despite the presence of paternal MHC histocompatibility antigens has been explained in mice by a Th2 switch at the level of the materno-fetal interface. We showed that some hormones enhanced during pregnancy can affect the development of Th1 and Th2 responses. Indeed, we found that progesterone promotes the production of IL-4 and IL-5, whereas relaxin promotes the production of IFN-gamma by T-cells. In addition, we showed that leukaemia inhibitory factor (LIF), which is essential for embryo implantation, associates with Th2 cells and is upregulated by IL-4 and progesterone. We also showed that LIF is down-regulated by Th1 inducers [IL-12, IFN-gamma and IFN-alpha]. Furthermore, we found a decreased production of LIF, IL 4 and IL-10 by decidual T-cells in women with unexplained recurrent abortions in comparison with women with normal gestation at the moment of voluntary abortion. The decreased production of LIF, IL-4 and IL-10 was not found in peripheral-blood T-cells. These results suggest that the local production of LIF and/or Th2 cytokines may contribute to the maintenance of pregnancy. PMID- 10816131 TI - Genetic analysis of insulin-like growth factor II and HLA-G in pre-eclampsia. AB - Pre-eclampsia (PE) is uniquely a disease of pregnancy and is the major cause of foetal and maternal morbidity and mortality. Epidemiological studies show that PE is highly heritable, with a high incidence in all populations. The underlying pathology indicates that absent or shallow invasion of foetal trophoblasts into maternal arteries is a feature of true PE. The objective of this study was to determine the genetic factors influencing PE. A large number of mother-father baby trios were collected in which the first pregnancy was complicated by severe PE. After careful examination of the epidemiology and pathology of the disease, two plausible candidate genes, namely insulin-like growth factor II (IGF-II) and HLA-G, were analysed for association with PE. No association was found between a commonly occurring polymorphism in IGF-II and PE. Three polymorphisms in HLA-G were analysed in the sample cohorts. No association was found between three polymorphisms in HLA-G and PE. However, the frequency of the HLA-G insertion/deletion polymorphism in exon 8 deviated significantly from Hardy Weinberg expectations in PE off-spring, reflecting an excess of heterozygotes for these polymorphisms in PE offspring. The significance of this deviation is not clear and further genetic analysis will be necessary to confirm this finding and to explore further the candidacy of HLA-G in PE. PMID- 10816132 TI - Cell senescence and human aging: where's the link? AB - Normal human cells will not divide forever in culture. After a defined number of passages, every culture enters a viable non-dividing state termed senescence. This led to the proposal that the progressive accumulation of senescent cells contributes to (but does not exclusively cause) the aging process. Data now suggest that cell senescence, like apoptosis, occurs as an anti-cancer mechanism. In vivo, extremely low rates of division occur over very long periods of time. Thus, by the end of its lifespan, an organism has performed a surprisingly large amount of cell turnover. In a normal culture the fraction of senescent cells increases smoothly with proliferation in vitro. This process can be accurately modelled by the progressive loss of telomeric sequence (proposed as a counting mechanism for senescence). Telomere length is usually maintained by the enzyme telomerase (repressed in many human tissues). Re-introduction of telomerase prevents the onset of senescence. However, data suggest that a separate telomere independent pathway exists. Evidence that this pathway is present in humans is now emerging. PMID- 10816134 TI - Regulation of cell cycle re-entry by growth, survival and stress signalling pathways. AB - The mitogen-activated and stress-activated protein kinases transduce signals from plasma membrane signalling machinery into the nucleus to modulate gene expression. By regulating the genomic response to environmental cues (growth factors, stresses) these pathways determine whether a cell re-enters the cell cycle, undergoes cell cycle arrest, senescence or apoptosis. We are particularly interested in how these pathways integrate with each other, and interact with the cell cycle machinery to achieve these discrete biological responses. PMID- 10816133 TI - Replicative senescence as a barrier to human cancer. AB - There is evidence that one critically short telomere may be recognized as DNA damage and, as a consequence, induce a p53/p21WAF- and p16INK4A-dependent G1 cell cycle checkpoint to cause senescence. Additionally, senescence via a p53- and p16(INK4A)-dependent mechanism can be induced by the over- or under-stimulation of certain signalling pathways that are involved in cancer. Central to this alternative senescence mechanism is the p14ARF protein, which connects oncogene activation, but not DNA damage, to p53 activation and senescence. We find that immortal keratinocytes almost invariably have dysfunctional p53 and p16 and have high levels of telomerase, but very often express a wild-type p14(ARF). Furthermore, when normal keratinocytes senesce they show a striking elevation of p16 protein, but not of p14(ARF) or its downstream targets p53 and p21(WAF). These results suggest that p16, rather than p14(ARF), is the more important gene in human keratinocyte senescence, but do not exclude a co-operative role for p14(ARF), perhaps in the induction of senescence by activated oncogenes in neoplasia. Regardless of mechanism, these results suggest that replicative senescence acts as a barrier to human cancer development. PMID- 10816135 TI - Telomere diminution as a cause of immune failure in old age: an unfashionable demurral. AB - The hypothesis that cellular proliferation leads to telomere shortening, which in turn leads to replicative failure, which in turn leads to a failure of immune function in aged individuals, is here evaluated against the published evidence about the nature and pace of immune decline in animals and humans. Although the evidence is strong that telomere shortening in late-passage human lymphocyte and non-lymphocytic cell lines induces a state in which the cells can no longer divide, there is no compelling evidence to suggest that replicative senescence of this kind is an important contributor to immune deficiency in old age. On the contrary, the accelerated pace of immune decline in mice and rats, whose telomeres are much longer than those of humans, argues strongly that the factors that pace age-dependent immune decline do not include telomere shortening. In addition, three subsidiary arguments - (a) the decline with age in naive T cell proliferation despite their relatively long telomeres; (b) the preservation of T cell proliferation in Werner's syndrome patients despite their cell lines' proclivity to replicative senescence in vitro; and (c) the ability of PMA and ionomycin to stimulate proliferation in T cells from old donors, but not in late passage T celt lines - all support the conclusion that aging of the immune system in living animals is not a consequence of the kind of replicative senescence typically caused by short telomeres in vitro. PMID- 10816136 TI - Measurement of telomere length in haematopoietic cells using in situ hybridization techniques. AB - The DNA of human chromosomes terminates in several kilobases of telomere repeats that are gradually lost with; age and with replication in vitro. Defective telomere maintenance has been shown to be causally linked to cell cycle exit and apoptosis. In order to overcome the limitations imposed by Southern blotting, we have established a quantitative fluorescence in situ hybridization (Q-FISH) technique. This technique allows estimation of telomere length in specific chromosome arms from metaphase cell preparations. Furthermore, we have extended quantitative in situ hybridization to flow cytometry (flow FISH) in order to obtain information on the mean telomere repeat content in suspended cells. Telomere length in granulocytes, monocytes, CD8 and CD4 T lymphocytes and natural killer cells was found to differ slightly in the peripheral blood of adults. However, strikingly longer telomeres were observed in B lymphocytes (approximately 1.3 kb longer), suggesting a functional role for telomere maintenance in this cell subset. In summary, Q-FISH and flow FISH represent new methods for measuring telomere length in single cells and allow studies of telomere dynamics in haematopoietic subpopulations at various stages of normal and abnormal antigen responses. PMID- 10816137 TI - Immunosenescence: potential causes and strategies for reversal. AB - Age-related deterioration in immune function has been recognized in many species. In humans the clinical manifestation of such immune dysfunction is age-related increases in the susceptibility to certain infections and in the incidence of some autoimmune disease and certain cancers. Laboratory investigations reveal age related changes in the peripheral T cell pool, in the predominant phenotype, cytokine production profiles, signalling function and in replicative ability following stimulus with antigen, mitogens or anti-CD3 antibody. These changes in the properties of peripheral T cells are thought to be causally linked to an age associated involution in the thymus. Our analysis reveals that thymic involution is due to a change in the thymic microenvironment linked to a reduction in the level of available interleukin 7. Treatment with interleukin 7 leads to a reversal of thymic atrophy with increased thymopoiesis. This provides the potential to reverse the immune dysfunction seen in the peripheral T cell pool by replacing old cells with new output generated in the thymus. Problems to overcome in order for such an experimental therapy to be successful require careful analysis in order to provide an optimal strategy to ensure that new T cell emigrants from the thymus have a broad range of specificities and are able to enter the peripheral T cell pool. PMID- 10816138 TI - Regulation of apoptosis and replicative senescence in CD8+ T cells from patients with viral infections. AB - Activated T lymphocytes are generated during an immune response. The induction of T lymphocyte proliferation is one way in which cell numbers can be controlled. However, once generated, the increased numbers of cells must be removed in order to re-establish cellular homoeostasis within the immune system. In this paper we describe how the numbers of activated T cells can be regulated by two distinct mechanisms, namely apoptosis and replicative senescence. In addition, we suggest that the regulation of cell clearance, as opposed to cell persistence, after an immune response is intimately involved in the generation of immune memory. PMID- 10816139 TI - Evidence in favour of ancient octaploidy in the vertebrate genome. AB - Vertebrate genomes are larger than invertebrates and show evidence of extensive gene duplication, including many collinear chromosomal segments. On the basis of this intra-genomic synteny, it has been proposed that two rounds of whole genome duplication (octaploidy) occurred early in the vertebrate lineage. Recently, this early vertebrate octaploidy has been challenged on the basis of gene trees. We report new linkage groups encompassing the matrilin (MATN), syndecan (SDC), Eyes Absent (EYA), HCK kinase and SRC kinase paralogous gene quartets. In contrast to other studies, the sequence trees are weakly supportive of ancient octaploidy. It is concluded that there is no strong evidence against the octaploidy, provided that consecutive genome duplication was rapid. PMID- 10816140 TI - Searching for the ideal forms of proteins. AB - A modification of the Structure Alignment Program (SAP), combined with a novel automatic method for the definition of structural elements, correctly identified the core folds of a variety of small beta/alpha proteins when compared with a series of ideal architectures. This approach opens the possibility of not just determining whether one structure is like another, but given a range of ideal forms, determining what the protein is. Preliminary studies have shown it to work equally well on the all alpha-class and the all-beta class of protein, each of which have corresponding ideal forms. Given the speed of the algorithm, it will be possible to compare all of these against the Protein Structure Database and determine the extent to which the current ideal forms can account for the variety of protein structure. Analysis of the remainder should provide a base for the development of further forms. PMID- 10816141 TI - Using the CATH domain database to assign structures and functions to the genome sequences. AB - The CATH database of protein structures contains approximately 18000 domains organized according to their (C)lass, (A)rchitecture, (T)opology and (H)omologous superfamily. Relationships between evolutionary related structures (homologues) within the database have been used to test the sensitivity of various sequence search methods in order to identify relatives in Genbank and other sequence databases. Subsequent application of the most sensitive and efficient algorithms, gapped blast and the profile based method, Position Specific Iterated Basic Local Alignment Tool (PSI-BLAST), could be used to assign structural data to between 22 and 36 % of microbial genomes in order to improve functional annotation and enhance understanding of biological mechanism. However, on a cautionary note, an analysis of functional conservation within fold groups and homologous superfamilies in the CATH database, revealed that whilst function was conserved in nearly 55% of enzyme families, function had diverged considerably, in some highly populated families. In these families, functional properties should be inherited far more cautiously and the probable effects of substitutions in key functional residues carefully assessed. PMID- 10816142 TI - Origins and evolution of AIDS viruses: estimating the time-scale. AB - The primate lentiviruses comprise SIV strains from various host species, as well as two viruses, HIV-1 and HIV-2, that cause AIDS in humans. The origins of HIV-1 and HIV-2 have been traced to cross-species transmissions from chimpanzees and sooty mangabey monkeys respectively. Two approaches have been taken to estimate the time-scale of the evolution of these viruses. Certain groups of SIV strains appear to have evolved in a host-dependent manner, implying a time-scale of many thousands or even millions of years. In stark contrast, molecular clock calculations have previously been used to estimate a time-scale of only tens or hundreds of years. Those calculations largely ignored heterogeneity of evolutionary rates across different sites within sequences. In fact, the distribution of rates at different sites seems extremely skewed in HIV-1, and so the time-depth of the primate lentivirus evolutionary tree may have been underestimated by at least a factor of ten. However, these date estimates still seem to be far too recent to be consistent with host-dependent evolution. PMID- 10816143 TI - Refractory mucosal candidiasis in advanced human immunodeficiency virus infection. AB - We conducted a multicenter, prospective study of the risk factors, natural history, and outcome of fluconazole-refractory mucosal candidiasis (FRMC) in 832 persons with advanced human immunodeficiency virus (HIV) infection (median CD4 cell count, 14/mm3) during 1994-1996. FRMC was defined as mucosal candidiasis that failed to resolve despite 14 days of therapy with daily doses (> or =200 mg) of fluconazole. Thirty-six persons (4.3%) had FRMC (35, oral; 1, esophageal), for an incidence of 4.2 per 100 person-years (859.7 total years of follow-up). In a multivariate model, the use of trimethoprim-sulfamethoxazole within 6 months of enrollment (relative risk [RR], 2.39; P=.04) and the use of fluconazole daily or every other day (RR, 5.64; P=.004) were significantly associated with the development of FRMC. The median survival after the development of FRMC was 32.6 weeks. In conclusion, the annual incidence of FRMC was <5%. Refractory candidiasis was a poor prognostic indicator. Daily or every-other-day use of fluconazole was associated with the development of refractory infection. PMID- 10816144 TI - Value of long-term administration of acyclovir and similar agents for protecting against AIDS-related lymphoma: case-control and historical cohort studies. AB - Acyclovir or similar agents with activity against Epstein-Barr virus (EBV) theoretically may prevent non-Hodgkin's lymphoma (NHL) in AIDS. A case-control study of 29 patients with AIDS-related NHL and 58 matched control subjects assessed the frequency with which daily acyclovir (>/=800 mg/d) or similar agents were used for > or =1 year. In a historical cohort of 304 patients with AIDS for > or =2 years, the prevalence of NHL was assessed among 3 groups of patients: those who received long-term treatment with high-dose acyclovir (or similar agents) or low-dose or intermittent acyclovir; those treated with ganciclovir/foscarnet for <1 year; and those who had not previously been treated with acyclovir, ganciclovir, or foscarnet. In the case-control study, 22 patients (72.4%) with NHL never received acyclovir or similar drugs versus 19 control subjects (32.8%; P=. 002); 2 patients (6.9%) with NHL received acyclovir (> or =800 mg/d) for > or =1 year versus 27 (46.6%) of control subjects (P=.0001). In the cohort study, 6 (6.8%) of 88 patients who received acyclovir (> or =800 mg/d) for > or =1 year developed NHL versus 15 (15.5%) of 97 patients who received intermittent or lower-dose acyclovir and 30 (25.2%) of 119 patients who never received these agents (P=.002). Long-term administration (>1 year) of high-dose acyclovir or similar agents with anti-EBV activity may prevent NHL in patients with AIDS. A prospective, randomized study is warranted to confirm these results. PMID- 10816145 TI - The need for investigations of prophylactic regimens to prevent AIDS-associated non-Hodgkin's lymphoma. PMID- 10816146 TI - Bronchopulmonary and mediastinal leishmaniasis: an unusual clinical presentation of Leishmania donovani infection. AB - We describe a case of unusual leishmaniasis in a Sudanese man with a history of progressively enlarging granulomatous mediastinal lymphadenopathy, worsening hemoptysis, and an intense mucosal granulomatous inflammatory response in the large bronchi. Leishmania donovani DNA was detected in bronchial biopsies by polymerase chain reaction. This is a novel description of human leishmanial infection in an immunocompetent patient involving this anatomical site. The patient's condition improved clinically, spirometrically, and radiologically after a course of treatment with amphotericin B. The cell-mediated immune response was analyzed before, during, and after successful antileishmanial chemotherapy. PMID- 10816147 TI - Enteropathogens in adult patients with diarrhea and healthy control subjects: a 1 year prospective study in a Swedish clinic for infectious diseases. AB - A 1-year prospective study was conducted to identify enteropathogens in adults with diarrhea (n=851) and in healthy control subjects (n=203) by use of conventional laboratory methods. Virulence factor genes for diarrheagenic Escherichia coli were detected by polymerase chain reaction. Enteropathogens were identified in 56% of patients and 16% of control subjects. The isolation rate was 65% for patients with symptoms for <1 week and for travelers; >1 pathogen was found in 11% of patients. The most frequent enteropathogens were Campylobacter (13% of patients), Clostridium difficile (13%), enterotoxigenic Escherichia coli (8%), Salmonella (7%), Shigella (4%), Blastocystis hominis (4%), calicivirus (3%), rotavirus (3%), enteroaggregative E. coli (2%), Aeromonas (2%), Giardia intestinalis (2%), Cryptosporidium (2%), and astrovirus (2%). Less frequently isolated (< or =1% of patients) were verotoxigenic E. coli, enteropathogenic E. coli, enteroinvasive E. coli, Entamoeba histolytica/Entamoeba dispar, microsporidia, and adenovirus. Fifty percent of the patients were hospitalized, and 43% needed intravenous fluids. The median duration of diarrhea was 14 days. Clinical features were not helpful for predicting the etiology of diarrhea. PMID- 10816149 TI - Severe pneumococcal pneumonia in previously healthy children: the role of preceding influenza infection. AB - An outbreak of severe pneumococcal pneumonia among children occurred in Iowa from November 1995 through January 1996. An associated outbreak of influenza disease was predominantly caused by influenza A (H1N1) for the first time since 1989. We conducted a case-control study to determine whether preceding influenza infection was directly associated with pneumococcal illness. We identified 13 children with severe pneumococcal pneumonia. Patients were more likely than control subjects to report experiencing an influenza-like illness in the 7-28 days preceding admission (matched odds ratio [OR], 12.4; 95% confidence interval [CI], 1.7-306). Likewise, family members of patients were more likely than those of control subjects to report experiencing an influenza-like illness in the 28 days preceding their admission date (OR, 2.6; 95% CI, 1.0-6. 3). Patients were more likely than control subjects to have a positive influenza A (H1N1) convalescent serology (matched OR, 3.7; 95% CI, 1.0-18.1). This study provides direct and indirect evidence that influenza infection led to severe pneumococcal pneumonia among these children. Prevention of pneumococcal disease should be included among the potential benefits of influenza vaccination. PMID- 10816148 TI - Use of rifabutin with protease inhibitors for human immunodeficiency virus infected patients with tuberculosis. AB - Drug interactions between rifamycins and highly active antiretroviral therapy (HAART) have raised concerns in the treatment of human immunodeficiency virus (HIV)-infected patients with tuberculosis. We conducted a study of this interaction by measuring serum drug levels of all HIV-infected patients with tuberculosis who were admitted to A. G. Holley State Tuberculosis Hospital (Florida) from October 1997 through December 1998, who were concomitantly treated with rifabutin and HAART. All 25 patients studied became culture-negative within 2 months of initiation of therapy for tuberculosis and remained negative for a median of 13 months follow-up after completion of therapy. HIV viral loads (mean+/-SEM) decreased significantly from 4.95+/-0.21 log10 copies/mL before initiation of HAART to 2.77+/-0.07 log10 copies/mL before discharge (P<.001); 20 of 25 patients achieved viral loads of <500copies/mL. In summary, the concomitant use of rifabutin and HAART can lead to successful treatment of HIV-infected patients with tuberculosis without increased side effects. PMID- 10816150 TI - Antimicrobial susceptibility testing: special needs for fastidious organisms and difficult-to-detect resistance mechanisms. AB - Clinical microbiology laboratories are faced with the challenge of accurately detecting emerging antibiotic resistance among a number of bacterial pathogens. In recent years, vancomycin resistance among enterococci has become prevalent, as has penicillin resistance and multidrug resistance in pneumococci. More recently, strains of methicillin-resistant Staphylococcus aureus with reduced susceptibility to vancomycin have been encountered. In addition, molecular techniques have demonstrated that there are still problems detecting methicillin resistance in staphylococci, especially in coagulase-negative species. Among members of the family Enterobacteriaceae, mutated beta-lactamase enzymes may confer difficult-to-detect resistance to later-generation penicillins and cephalosporins. Anaerobic bacteria are no longer entirely predictable in their susceptibility to agents that might be selected for empiric therapy. Therefore, clinical microbiology laboratories may not be able to rely on a single susceptibility testing method or system to detect all those emerging resistant or fastidious organisms. For reliable detection, laboratories may need to employ conventional, quantitative susceptibility testing methods or use specially developed, single concentration agar screening tests for some resistant species. Certain of these screening tests are highly specific, while others may require additional confirmatory testing for definitive results. Therefore, laboratories must retain the versatility to apply several different approaches to detect resistance in both common and infrequently encountered bacterial pathogens. PMID- 10816151 TI - Treatment of cutaneous larva migrans. AB - Cutaneous larva migrans caused by the larvae of animal hookworms is the most frequent skin disease among travelers returning from tropical countries. Complications (impetigo and allergic reactions), together with the intense pruritus and the significant duration of the disease, make treatment mandatory. Freezing the leading edge of the skin track rarely works. Topical treatment of the affected area with 10%-15% thiabendazole solution or ointment has limited value for multiple lesions and hookworm folliculitis, and requires applications 3 times a day for at least 15 days. Oral thiabendazole is poorly effective when given as a single dose (cure rate, 68%-84%) and is less well tolerated than either albendazole or ivermectin. Treatment with a single 400-mg oral dose of albendazole gives cure rates of 46%-100%; a single 12-mg oral dose of ivermectin gives cure rates of 81%-100%. PMID- 10816152 TI - Human immunodeficiency virus type 1 test results in patients with malaria and dengue infections. AB - Six rapid enzyme immunoassays for the detection of HIV antibody were performed on paired sera from 66 patients with malaria and 9 patients with dengue. Kit specificities ranged from 77% to 100%, demonstrating that more data are needed on cross-reactivity with endemic diseases as the use of rapid HIV tests increases. PMID- 10816154 TI - Mycotic aneurysm of the abdominal aorta in a patient undergoing hemodialysis: an unusual complication of Staphylococcus aureus bacteremia. AB - When Staphylococcus aureus is repeatedly positive in blood cultures even under effective antibiotics therapy (vancomycin, teicoplanin, or rifampin), computed tomography scan and sonography should be performed early to exclude mycotic aneurysm of the deeply seated arteries, especially in patients with abdominal aortic calcification. Before 1990, the most common causative organism of suprarenal aortic mycotic aneurysm was Salmonella; since 1990, it has been gram positive cocci (i.e., Streptococcus and Staphylococcus) rather than gram-negative bacilli (i.e., Salmonella), possibly because of the more invasive procedures performed in clinical settings, but this hypothesis needs further investigation. PMID- 10816153 TI - Recurrent subcutaneous infection due to Scopulariopsis brevicaulis in a liver transplant recipient. AB - We describe a case of recurrent Scopulariopsis brevicaulis subcutaneous infection, which occurred 6 years after the patient underwent liver transplantation. Combined surgery and long-term oral therapy with terbinafine resulted in a favorable outcome, although this is not the rule in the previously reported S. brevicaulis infections in immunocompromised patients. PMID- 10816155 TI - Diagnostic accuracy of transthoracic and multiplane transesophageal echocardiography for valvular perforation in acute infective endocarditis: correlation with anatomic findings. AB - We evaluated the diagnostic accuracy of transthoracic and multiplane transesophageal echocardiography (TTE and TEE, respectively) for assessing valvular perforation during active infective endocarditis by correlating the results of TTE and TEE with anatomic findings of 88 valves examined at surgery or autopsy. Compared with TEE, TTE has a low diagnostic sensitivity in the detection of this complication and, in the presence of hemodynamic instability, multiplane TEE should be performed directly. PMID- 10816156 TI - Inflammatory pseudotumor causing small bowel obstruction and mimicking lymphoma in a patient with AIDS: clinical improvement after initiation of thalidomide treatment. AB - A patient with AIDS was diagnosed with inflammatory pseudotumor with small bowel involvement. After receiving thalidomide treatment, serum tumor necrosis factor (TNF) and soluble TNF receptor II levels normalized, his constitutional and gastrointestinal symptoms diminished, and the mass lesion shrunk. PMID- 10816157 TI - Disseminated Mycobacterium terrae infection in a patient with advanced human immunodeficiency virus disease. AB - Mycobacterium terrae has been rarely implicated in human disease and never in patients infected with human immunodeficiency virus (HIV). We describe an HIV infected patient with disseminated infection by M. terrae with pulmonary and cutaneous clinical manifestations. M. terrae was isolated from both sputum and urine, and identified by both conventional tests and high-performance liquid chromatography. Clinical and microbiological characteristics of this case are compared with those reported in the literature. PMID- 10816158 TI - Diagnosing postneurosurgical meningitis. PMID- 10816159 TI - Neurological dysfunction following malaria: disease- or drug-related? PMID- 10816160 TI - Reply PMID- 10816161 TI - Reply PMID- 10816163 TI - Reply PMID- 10816162 TI - Impact of infectious diseases specialists. PMID- 10816164 TI - Herpes simplex virus peritonitis. PMID- 10816165 TI - Report of the first cases of cutaneous leishmaniasis in East Timor. PMID- 10816167 TI - [Left ventricular diastolic dysfunction in young people with type 1 diabetes mellitus. Associated factors]. AB - AIM: The aim of our study was to evaluate left ventricular function of diastolic in young (< 40 years) asymptomatic patients with type 1 diabetes mellitus free of cardiovascular disease symptoms and to analyze the associated factors to the left ventricular diastolic dysfunction (LVDD) in these patients. PATIENTS AND METHODS: Thirty-five type-1 diabetic patients (mean age 27.8+/-7.5 years) old and 54 healthy controls (mean age 26.1+/-4.1 years) were studied. Anamnesis, physical exploration, general analytical studies, microalbuminuric and Doppler echocardiographic studies were performed. RESULTS: The LVDD was present in 13 (37.1%) of the diabetic patients and none of the control patients. The ratio of peak early to peak late (atrial) filling velocity was significantly decreased in diabetic compared with control subjects (1.1+/-0.3 versus 1.5+/-0.2; p<0.01). The isovolumetric relaxation time was increased in diabetic patients compared with control subjects (104+/-11 versus 79+/-11; p<0.01). Diabetics with LVDD were older aged, predominantly males, had worse glucemic control, more alteration of lipidic metabolism and higher levels of microalbuminuria, than diabetics without LVDD. CONCLUSIONS: The LVDD is frequent in young diabetics free of cardiovascular disease symptoms. These studies suggest that because this patients were of older age, of the masculine sex with, poor glucemic control, altered lipidic metabolism, and microalbuminuria they might be a group that is associated with LVDD which, in the absence of cardiovascular disease, might be an early preclinical alteration, potentially related to subsequent development of diabetic cardiomyopathy. PMID- 10816168 TI - [Diagnostic accuracy of tomographic myocardial imaging for evaluation of aortocoronary graft patency]. AB - INTRODUCTION AND OBJECTIVES: Conventional ergometry has limitations in the evaluation of coronary artery bypass graft patency. The aim of the present study was to determine the efficacy of exercise single photon emission tomography with 99mTc-compounds for the diagnosis of coronary artery bypass grafts disease. PATIENTS AND METHODS: The state of sixty-seven coronary artery bypass grafts (31 with mammary artery, 36 with safein vein; 35 to left descending anterior artery, 15 to circumflex and 17 to right coronary artery) were analyzed retrospectively in 38 patients (mean age 63+/-8.7; 35 men). The time that elapsed between the coronary surgery and the exercise tomography was 9,7 years. In 16 cases, exercise tomography was performed with 99mTc-MIBI and in the 22 remaining with 99mTc tetrofosmin. In 6 cases, dipiridamol was administred simultaneously during an insufficient exercise test. RESULTS: Sensitivity (73.1%), specificity (93%), positive predictive value (86.3%), negative predictive value (84.4%), global value (85%), positive likelihood ratio (10.4) and negative likelihood ratio (0.29) of exercise tomography were significantly (p<0.01) better than those obtained with the exercise test alone (53.8%, 43.6%, 38.9%, 58. 6%, 47.7%, 0.95 and 1.06, respectively). CONCLUSIONS: Exercise tomography with 99mTc-compounds is a highly effective for the detection of coronary artery bypass grafts disease. PMID- 10816169 TI - [Determinants of contractile reserve in the infarction area. A quantitative study using dobutamine in contrast left ventriculography]. AB - AIM: The aim of this study was to relate the contractile reserve in infarction segments to the dysfunction at rest and to the residual coronary stenosis. METHODS: The study group consisted of 95 patients with a first myocardial infarction. Contrast left ventricular at baseline and after dobutamine infusion at 7.5 microg/kg/min and coronary angiograms were performed. The centerline method was used to quantify the extent of dysfunction (percentage of chords with dysfunction in the territory of the infarction artery) and its maximum severity (maximum units of standard deviation [SD] below the normal wall motion reference). Reduction of dysfunction extent with dobutamine was measured. RESULTS: On increasing baseline dysfunction severity, both the magnitude of the response to dobutamine ( 2 SD 3 SD 4 SD +/- 5 SD [n = 15] = 9+/-13%, > 5 SD [n = 13] = 3+/-4%, p = 0,0001), and the number of patients with a significant (> or =15%) positive response ( 2 SD 3 SD 4 SD 5 SD = 0%, p<0,0001) decreased. There were no differences in dobutamine improvement among the subgroups with (n = 84) or without (n = 11) significant stenosis in the infarction artery (18+/-15 vs. 16 +/-18%), or between the subgroups with a patent (n = 76, 18+/-19%) or occluded (n = 19, 11+/-11%) artery. CONCLUSIONS: Dobutamine response is related to dysfunction severity in the infarction area: when the severity is 5 (high negative response prevalence), dobutamine testing does not seem indicate. The existence of residual coronary stenosis does not attenuate contractile reserve at low dobutamine doses. PMID- 10816170 TI - [Short and long-term prognosis of prosthetic valve endocarditis in non-drug addicts]. AB - INTRODUCTION AND OBJECTIVES: Prosthetic valve infective endocarditis is a complication of valvular replacement surgery with a high morbimortality during the in-hospital phase and an important risk of complications during follow-up. The objective of the present study is to assess the clinical features and the short and long-term prognosis of this disease. PATIENTS AND METHODS: A prospective study of 43 consecutive cases of prosthetic valve endocarditis in non addict patients from January 1987 to March 1997. RESULTS: The mean age was 51 +/ 16 years. Eight patients (19%) had early prosthetic valve endocarditis (two months following heart surgery), fourteen patients (32%) had intermediate (between 2 and 12 months post surgery) and twenty-one (49%) had late prosthetic valve endocarditis (more than one year after heart surgery). Transesophageal echocardiography was performed in 32 patients with a sensibility of 81%. Complications occurred in 86% of patients and 53% of patients underwent surgery during the active phase (25% was emergency surgery). Inpatient mortality was 23% (50% in early prosthetic valve endocarditis). After a mean follow-up of 56 months there were 5 cases of recurrence, four patients required late surgery and 5 patients died. Survival (excluding early mortality) was 82% at 5 years with no significant differences among patients who received only medical treatment and those who underwent surgery in the active phase. CONCLUSIONS: Early mortality of prosthetic valve endocarditis is, according to our experience of 20%. The prognosis of survivors to the active phase is favourable in the majority. Early prosthetic valve endocarditis still causes a high mortality rate despite the use of combined medical surgical treatment in most cases. PMID- 10816171 TI - [Total arterial myocardial revascularization with both mammary arteries without extracorporeal circulation]. AB - INTRODUCTION: Tector has described the off-pump total arterial revascularization technique, using multiple anastomosis with both internal thoracic arteries. To reduce surgical morbid-mortality, we have proposed the use of this technique without extracorporeal circulation. PATIENTS AND METHODS: From April, 1998 the off-pump <> technique was performed in 92 patients, 74 male (80%) and 18 female (20%), with a mean age of 64.9+/-8.1 years (42-78). Preoperative angiography demonstrated triple-vessel disease in 58 (63%) patients, and left main disease was present in 19 (20.5%) patients. Forty patients (43.5%) showed unstable angina, 24 patients (26%) significant peripheral vascular disease, and 26 (28%) diabetes mellitus. Both internal thoracic arteries were harvested using the skeletonization technique and were used like a <> graft. The flow in the graft was measured using a flowmeter, and in 24 (26%) patients by angiographic study. RESULTS: A total of 274 distal anastomoses were performed, 122 (44.6%) in the lateral or inferior wall, and 69 (25.2%) were sequential, with an average of 2.98 bypass/patient. In 59.8% of the patients a triple bypass was performed, 22% double bypass, 17% cuadruple bypass and 1 patient a quintuple bypass. During the initial six hours 64.9% of patients were extubated. Only one patient (1.1%) needed intraaortic ballon pumping and 3 (3.2%) inotropics during the postoperative course. Hospital mortality was 3 (3.2%) patients. Reoperation for bleeding was needed in just one patient (1.1%), and 78.3% of patients were not transfused. Mediastinitis occurred in 3 patients (3.2%). Postoperative stroke was not observed. At 7.7+/-2.8 months of mean follow-up all patients were free of symptoms and the global patency rate of 94%. CONCLUSIONS: Off-pump Tector technique appears to be safe, offering a complete arterial revascularization and showing a reduction of surgical morbidity. PMID- 10816172 TI - [Radiofrequency ablation of idiopathic left ventricular tachycardia in children]. AB - INTRODUCTION: Nowadays radiofrequency catheter ablation is an alternative to medical treatment in adult patients with idiopathic left ventricular tachycardia. AIM: To asses indications, results and long term follow-up of using this technique in children. : Radiofrequency catheter ablation was performed in five consecutive patients without structural heart disease, mean age 8.6+/-7.1 years (range, 1.3 to 17) and suffering from resistance to medical treatment left ventricular tachycardia (2.4+/-0.9 antiarrhythmic drugs per patient). Palpitations and syncope were the clinical manifestations in four patients while congestive heart failure in the other one. Six types of tachycardia were identified in the five-patient group. All of them responded to intravenous verapamil. There were 3 incessant tachycardias and the other three were paroxysmal ones. The ablation site was selected using activation mapping during tachycardia and pace-mapping. The Tachycardia cycle length was 275+/-123 ms and showed right bundle branch block with superior axis (left in 4 and right in 2 cases). The ablation target was located in the mid-septal region of the left ventricle in 3 cases, in the apical and inferior septal region in 2 other cases and in the anterior free wall in the other one. Local activation times preceded in 33+/-13 ms at QRS beginning and with a 12/12 agreement pattern in 5 cases and 9/12 in 1, in whom ablation was unsuccessful. At follow-up, the 4 patients with effective ablation have been tachycardia-free for 35.8+/-17 months. CONCLUSIONS: Radiofrequency catheter ablation is a useful method of treatment in children with idiopathic left ventricular tachycardia, including those under two years old. PMID- 10816173 TI - [Clinical trials presented at the 49th Annual Scientific Session of the American College of Cardiology (Anaheim, March 2000). Abstracts]. PMID- 10816174 TI - [Clinical practice guidelines of the Spanish Society of Cardiology on echocardiography]. AB - Doppler echocardiography has become the technique of choice for the diagnosis and follow-up of most heart diseases its main advantages are that it is non-invasive, easy to use, readily available, rapid and has a high cost-benefict ratio. Multiple studies have demonstrated the accuracy of the technique in the diagnosis and severity quantification of the severity of different diseases. Nevertheless, its main limitation lies in the fact that results are operator dependent, and therefore correct training of the echocardiographer is mandatory. In addition, adequate infrastructure and appropriate equipment are a required guarantee quality of the study. Finally, the technique should be used for the indications, in which beneficial information can be yielded. The aim of the present article was to define and update these considerations to enhance the usefulness of echocardiography in clinical practice. PMID- 10816175 TI - [Clinical practice guidelines of the Spanish Society of Cardiology for physical activity in patients with cardiac disease]. AB - This chapter includes general recommendations regarding the eligibility for recreational and competitive sports participation. Although based on the latest knowledge in sports medicine and cardiology, these recommendations should be updated according to advances in the understanding of the athlete's heart. Physical exercise plays an important role in the leisure time of our society. Many patients with cardiac disease don't want to give up their physical or sport activity programs. Moreover, there is no doubt that many cardiovascular abnormalities may benefit from a controlled physical exercise program. Understanding the cardiac adaptations to exercise training, individualised evaluation of each cardiac pathology and adequate exercise prescription is essential in order to minimize the risks for every cardiac patient. PMID- 10816176 TI - [Evaluation and treatment of cardiac injuries]. AB - Cardiac injuries caused by a heart traumatism are not frequent but, of great importance given their high morbidity. Two different groups in terms of etiology, clinical picture, application of diagnostic techniques, treatment and prognosis can be considered. On one hand, there are cardiac injuries caused by a thoracal contusion, which provokes a contused lesion can affect the free wall, the interventricular septum, the valves, the subvalvular apparatus, the conduction system and the coronary vessels and, on the other hand, cardiac injuries caused by penetrating objects. Cardiac injury can lead to a life-threatening hemodynamic instability which mandates prompt and clear diagnostic and therapeutic approaches. PMID- 10816177 TI - [Prevention of sudden death in patients awaiting heart transplantation]. AB - Sudden death, unexpectedly alters outcome in many patients awaiting heart transplantation. The prevention of sudden death in these patients has been the focus of intensive research to achieve a larger number of patients who finally receive transplants. Recent advances in the medical treatment of heart failure, have reduced mortality and in particular, that caused by sudden death. Nonetheless sudden death remains a frequent cause of mortality in patients awaiting cardiac transplantation. The recognition of patients at very high risk for sudden death is relatively easy, but most patients who suffer sudden death while awaiting cardiac transplantation, are not among those initially included in the overall high risk category. The betablockers, when patients are able to use them, can reduce sudden and total mortality. Class I antiarrhythmic drugs should not be used in patients with cardiac failure. Amiodarone does not increase mortality and may have a beneficial effect in some patients, but its efficacy is lower than that of the implantable defibrillator and its widespread use is not justified. The implantable defibrilator is the reference treatment to reduce sudden death in selected patients, awaiting transplantation. PMID- 10816178 TI - [Chronic constrictive pericarditis secondary to thoracic traumatic hematoma]. PMID- 10816179 TI - [Demonstrated myocardial ischemia due to multiple coronary fistulae draining into the left ventricle]. AB - There are few cases of demonstrated ischemia due to the presence of multiple coronary fistulae. We report a case of multiple coronary fistulae from left anterior descending, circumflex and right coronary arteries draining into the left ventricle, which were responsible for myocardial ischemia demonstrated with thallium stress test. They were diagnosed by coronary angiography, and their etiology seems to be congenital. The final control of symptoms was achieved with beta-blockers, and not with nitrates, which speaks for a steal phenomenon as the cause of ischemia. PMID- 10816180 TI - [Perforating epicardial lead: atypical evolution]. AB - Endocarditis related to pacemaker lead is a rare complication of permanent transvenous pacing, of which the diagnosis is carried out with the presence of verrucae in echocardiography and positive blood cultures, its treatment being mixed -medical and surgical- because the isolated medical treatment is rarely successful and the lead should be extracted. We present the case of recurrent endocarditis of several years of evolution, in the which it was not possible to extract of the electrode due to the special characteristics of the patiente (epicardial lead perforating into right atrial). PMID- 10816181 TI - [Recurrent ventricular fibrillation during a febrile illness in a patient with the Brugada syndrome]. AB - Different situations have been involved in the origin of ventricular arrhythmic events in patients with the Brugada syndrome such as bradycardia, alcohol consumption and mental stress. We present a 30 year old male with recurrent ventricular fibrillation due to a febrile illness with intense sweating. He had been previously studied at our Unit in 1995 because of an episode of resuscitated cardiac arrest due to ventricular fibrillation. The twelve-lead electrocardiogram showed the typical characteristics of a patient with the Brugada syndrome. Different invasive and non-invasive tests performed were normal. He received a defibrillator and had no recurrences during 4 years of follow up. In March,1999, after an upper respiratory tract infection he had high fever treated with paracetamol but at down he had sweating and chills, followed by 3 defibrillator shocks. Late interrogation showed 5 episodes of ventricular fibrillation, two of them non-sustained, and the rest adequately treated by the defibrillator. Activation and inactivation kinetics for early INa are twofold faster at higher temperature, and shift activation and steady-state inactivation. This may explain the role of the temperature as a trigger for ventricular arrhythmias in our patient. PMID- 10816182 TI - [Use of intravenous esmolol in patient with severe heart failure and abnormal intraventricular gradient after aortic valve replacement]. AB - The presence of anomalous intraventricular flows of high velocity and dynamic ejective obstruction of the left ventricle post aortic valve replacement, generally by severe aortic stenosis, is a relatively infrequent, but severe complication. An early diagnosis is of crucial importance, since the usual treatments applied to postoperative heart failure are often inefficacious or even harmful. Even in cases of severe pump failure, when the clinical case is diagnosed adequately, endovenous short-action beta-blockers are useful. PMID- 10816183 TI - A comparison of nefazodone, the cognitive behavioral-analysis system of psychotherapy, and their combination for the treatment of chronic depression. AB - BACKGROUND: Patients with chronic forms of major depression are difficult to treat, and the relative efficacy of medications and psychotherapy is uncertain. METHODS: We randomly assigned 681 adults with a chronic nonpsychotic major depressive disorder to 12 weeks of outpatient treatment with nefazodone (maximal dose, 600 mg per day), the cognitive behavioral-analysis system of psychotherapy (16 to 20 sessions), or both. At base line, all patients had scores of at least 20 on the 24-item Hamilton Rating Scale for Depression (indicating clinically significant depression). Remission was defined as a score of 8 or less at weeks 10 and 12. For patients who did not have remission, a satisfactory response was defined as a reduction in the score by at least 50 percent from base line and a score of 15 or less. Raters were unaware of the patients' treatment assignments. RESULTS: Of the 681 patients, 662 attended at least one treatment session and were included in the analysis of response. The overall rate of response (both remission and satisfactory response) was 48 percent in both the nefazodone group and in the psychotherapy group, as compared with 73 percent in the combined treatment group. (P<0.001 for both comparisons). Among the 519 subjects who completed the study, the rates of response were 55 percent in the nefazodone group and 52 percent in the psychotherapy group, as compared with 85 percent in the combined-treatment group (P<0.001 for both comparisons). The rates of withdrawal were similar in the three groups. Adverse events in the nefazodone group were consistent with the known side effects of the drug (e.g., headache, somnolence, dry mouth, nausea, and dizziness). CONCLUSIONS: Although about half of patients with chronic forms of major depression have a response to short-term treatment with either nefazodone or a cognitive behavioral-analysis system of psychotherapy, the combination of the two is significantly more efficacious than either treatment alone. PMID- 10816184 TI - Daily interruption of sedative infusions in critically ill patients undergoing mechanical ventilation. AB - BACKGROUND: Conous infusions of sedative drugs in the intensive care unit may prolong the duration of mechanical ventilation, prolong the length of stay in the intensive care unit and the hospital, impede efforts to perform daily neurologic examinations, and increase the need for tests to assess alterations in mental status. Whether regular interruption of such infusions might accelerate recovery is not known. METHODS: We conducted a randomized, controlled trial involving 128 adult patients who were receiving mechanical ventilation and continuous infusions of sedative drugs in a medical intensive care unit. In the intervention group, the sedative infusions were interrupted until the patients were awake, on a daily basis; in the control group, the infusions were interrupted only at the discretion of the clinicians in the intensive care unit. RESULTS: The median duration of mechanical ventilation was 4.9 days in the intervention group, as compared with 7.3 days in the control group (P=0.004), and the median length of stay in the intensive care unit was 6.4 days as compared with 9.9 days, respectively (P=0.02). Six of the patients in the intervention group (9 percent) underwent diagnostic testing to assess changes in mental status, as compared with 16 of the patients in the control group (27 percent, P=0.02). Complications (e.g., removal of the endotracheal tube by the patient) occurred in three of the patients in the intervention group (4 percent) and four of the patients in the control group (7 percent, P=0.88). CONCLUSIONS: In patients who are receiving mechanical ventilation, daily interruption of sedative-drug infusions decreases the duration of mechanical ventilation and the length of stay in the intensive care unit. PMID- 10816185 TI - Coronary-artery calcification in young adults with end-stage renal disease who are undergoing dialysis. AB - BACKGROUND: Cardiovascular disease is common in older adults with end-stage renal disease who are undergoing regular dialysis, but little is known about the prevalence and extent of cardiovascular disease in children and young adults with end-stage renal disease. METHODS: We used electron-beam computed tomography (CT) to screen for coronary-artery calcification in 39 young patients with end-stage renal disease who were undergoing dialysis (mean [+/-SD] age, 19+/-7 years; range, 7 to 30) and 60 normal subjects 20 to 30 years of age. In those with evidence of calcification on CT scanning, we determined its extent. The results were correlated with the patients' clinical characteristics, serum calcium and phosphorus concentrations, and other biochemical variables. RESULTS: None of the 23 patients who were younger than 20 years of age had evidence of coronary-artery calcification, but it was present in 14 of the 16 patients who were 20 to 30 years old. Among those with calcification, the mean calcification score was 1157+/-1996, and the median score was 297. By contrast, only 3 of the 60 normal subjects had calcification. As compared with the patients without coronary-artery calcification, those with calcification were older (26+/-3 vs. 15+/-5 years, P<0.001) and had been undergoing dialysis for a longer period (14+/-5 vs. 4+/-4 years, P< 0.001). The mean serum phosphorus concentration, the mean calcium phosphorus ion product in serum, and the daily intake of calcium were higher among the patients with coronary-artery calcification. Among 10 patients with calcification who underwent follow-up CT scanning, the calcification score nearly doubled (from 125+/-104 to 249+/-216, P=0.02) over a mean period of 20+/-3 months. CONCLUSIONS: Coronary-artery calcification is common and progressive in young adults with end-stage renal disease who are undergoing dialysis. PMID- 10816186 TI - A five-year study of the incidence of dyskinesia in patients with early Parkinson's disease who were treated with ropinirole or levodopa. AB - BACKGROUND: There is debate about whether the initial treatment for patients with Parkinson's disease should be levodopa or a dopamine agonist. METHODS: In this prospective, randomized, double-blind study, we compared the safety and efficacy of the dopamine D2-receptor agonist ropinirole with that of levodopa over a period of five years in 268 patients with early Parkinson's disease. If symptoms were not adequately controlled by the assigned study medication, patients could receive supplementary levodopa, administered in an open-label fashion. The primary outcome measure was the occurrence of dyskinesia. RESULTS: Eighty-five of the 179 patients in the ropinirole group (47 percent) and 45 of the 89 patients in the levodopa group (51 percent) completed all five years of the study. In the ropinirole group 29 of the 85 patients (34 percent) received no levodopa supplementation. The analysis of the time to dyskinesia showed a significant difference in favor of ropinirole (hazard ratio for remaining free of dyskinesia, 2.82; 95 percent confidence interval, 1.78 to 4.44; P<0.001). At five years, the cumulative incidence of dyskinesia (excluding the three patients who had dyskinesia at base line), regardless of levodopa supplementation, was 20 percent (36 of 177 patients) in the ropinirole group and 45 percent (40 of 88 patients) in the levodopa group. There was no significant difference between the two groups in the mean change in scores for activities of daily living among those who completed the study. Adverse events led to the early withdrawal from the study of 48 of 179 patients in the ropinirole group (27 percent) and 29 of 89 patients in the levodopa group (33 percent). The mean (+/-SD) daily doses given by the end of the study were 16.5+/-6.6 mg of ropinirole (plus 427+/-221 mg of levodopa in patients who received supplementation) and 753+/-398 mg of levodopa (including supplements). CONCLUSIONS: Early Parkinson's disease can be managed successfully for up to five years with a reduced risk of dyskinesia by initiating treatment with ropinirole alone and supplementing it with levodopa if necessary. PMID- 10816187 TI - Images in clinical medicine. Mediastinal teratoma. PMID- 10816188 TI - Hypernatremia. PMID- 10816189 TI - Intrauterine infection and preterm delivery. PMID- 10816190 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 15-2000. A 69-year-old man with myasthenia gravis and a mediastinal mass. PMID- 10816191 TI - Is academic medicine for sale? PMID- 10816192 TI - Treatment of chronic depression. PMID- 10816193 TI - A wake-up call in the intensive care unit. PMID- 10816194 TI - Correction: Physicians' Experiences with the Oregon Death with Dignity Act. PMID- 10816195 TI - Correction: Cultivation of the Bacillus of Whipple's Disease. PMID- 10816196 TI - Uneasy alliance--clinical investigators and the pharmaceutical industry. PMID- 10816197 TI - Electrolyte and pH dependence of heart rate during hemodialysis: a computer model analysis. AB - The influence of hemodialysis-induced modifications in extracellular fluid characteristics on heart rate was investigated by using a detailed computer model of sinus-node electrical activity. Changes similar to those occurring in the course of hemodialysis in extracellular concentrations of sodium (from 138 to 140 mM), potassium (from 6 to 3.3 mM), and calcium (from 1.2 to 1.5 mM) ions as well as in pH (from 7.31 to 7.4) and intracellular volume were simulated. The model predicted that such changes may largely influence the rhythm of the sinoatrial node pacemaker, causing the heart rate to range from 69 to 86 bpm. Heart rate increases after removing potassium (up to 7 bpm) and also after calcium perfusion (up to 11 bpm) whereas restoring pH slows heart beat (up to 6 bpm). Extracellular sodium has no significant influence, but the heart rate strictly depends on intracellular sodium concentration (5 bpm/mM). A complex dependence of heart rate on electrolytes and pH was also recognized. Providing extracellular potassium concentration is maintained above 5 mM, heart rate exhibits low sensitivity to changes in calcium and potassium. When potassium concentration is reduced below 4.5 mM, heart rate sensitivity to calcium and potassium increases significantly to 10 and 30 bpm/mM, respectively. A sustained increase in heart rate always corresponds to an increase in intracellular sodium concentration. PMID- 10816198 TI - Natural changes in peritoneal equilibration test results in continuous ambulatory peritoneal dialysis patients: a retrospective, seven year cohort survey. AB - We conducted a retrospective, 7 year cohort survey to examine the natural changes in peritoneal equilibration test (PET) results in patients with long-term uneventful continuous ambulatory peritoneal dialysis (CAPD). Thirty-two (17 males, 15 females) patients on CAPD with two or more standard PETs performed more than 6 months apart, in the absence of peritoneal insult, were included. Changes and pattern of PET results were evaluated by the dialysate to plasma ratio of creatinine (D:P-cre), the fourth h dialysate to instilled glucose ratio (D4:Do) and ultrafiltration volume (UF, ml). The subgroups included high (H), high average (HA), low-average (LA), or low (L) transporters with the dividing ratios (D:P-cre) of >0.81, >0. 65 to 0.81, >0.5 to 0.65, and <0.5, respectively. The median D:P-cre significantly decreased (p = 0.04), but neither the D4:Do nor the final median UF significantly decreased. The change in D:P-cre was strongly and inversely correlated with the initial D:P-cre value (r = -0.68; p < 0.05). A similar relationship was found between the change in the final D4:Do and the initial D4:Do (r = -0.752; p < 0. 01) and between the change in the final UF and the initial UF (r = -0.875; p < 0.01). No correlation was found between the change in D:P-cre and the age of the patient, the time interval between PETs, monthly dialysate glucose exposure, or underlying diabetes/non-diabetes. The final peritoneal transport pattern was altered with 5 (15.6%) patients remaining in the extreme subgroups (H or L) and, by contrast, 84.4% (27/32) of the patients now in the averaged (HA or LA) groups (p < 0.01, chi2 test). We demonstrated a natural "centralization" migration of PET results after long-term uneventful CAPD, which may help to explain why patients with extreme PET characteristics, that is, H or L, continued to do well on CAPD. PMID- 10816199 TI - Comparative study of bioartificial liver support and plasma exchange for treatment of pigs with fulminant hepatic failure. AB - Recently, bioartificial liver (BAL) treatment was reported to provide beneficial effects for patients with fulminant hepatic failure (FHF). Some success in experimental or clinical trials has been reported; however, the evaluation of BAL efficacy remains unclear, especially in comparison with other treatments for FHF. The purpose of this study was to compare the efficacy between BAL and plasma exchange (PE) in experimentally induced FHF in pigs. Pigs undergoing hepatic devascularization (HD) were placed into the following groups: no treatment (control; n = 6), BAL treatment (BAL; n = 5), and plasma exchange (PE; n = 5). Each treatment was initiated 6 h after HD and lasted for 4 h. BAL treatment significantly improved liver functions in FHF pigs. The decrease in cerebral perfusion pressure was also significantly suppressed in the pigs with BAL, and their survival time was prolonged compared with the results in pigs with PE. The effects of BAL outperform those of PE in the treatment of experimental FHF model. PMID- 10816200 TI - Paper is a compatible bed for rat hepatocytes. AB - To develop an effective hybrid bioartificial liver (BAL) device, the material of the scaffold is very important to support hepatocytes that have both growth ability and hepatic differentiated functions. In this study we used paper (Kimwipe, Kimberly-Clark Corp., Roswell, GA, U.S.A.) as a scaffold. Primary hepatocytes isolated from a normal adult rat liver could proliferate on the paper. The secretion of albumin into culture medium by the cells on the paper increased with time in culture and, compared to the cells on dishes, the amount of 48 h albumin secretion at Day 10 was two times larger. Perpendicular sections of hepatocytes on the paper revealed that the cells fell into cavities made by intersecting fibers, piled up, and formed three to four layers. The piled-up cells changed their shape from flat to cuboidal and enlarged their cytoplasm, which was rich in organelles such as mitochondria and peroxisomes with a nucleoid. In addition, they formed bile canalicular structures between the cells. Their morphological appearance was similar to in vivo hepatocytes. Paper (Kimwipe) may be a good candidate as a scaffold to make a BAL device. PMID- 10816201 TI - Oxygen transfer in a diffusion-limited hollow fiber bioartificial liver. AB - A mathematical model was developed to predict oxygen transport in a hollow fiber bioartificial liver device. Model parameters were taken from the Hepatix ELAD configuration; a blood perfused hollow fiber cartridge with hepatocytes seeded in the extracapillary space. Cellular oxygen uptake is based on Michaelis-Menten kinetics, and nonlinear oxygen transport in the blood is considered. The effect of modulating three important parameters is investigated, namely, the Michaelis Menten constants Vm (volumetric oxygen consumption of the hepatocytes) and Km (half-saturation constant), and hollow fiber oxygen permeability. A computer implementation of the model is used to assess whether a given cell mass could be maintained within such a device. The results suggest that liver cell lines possessing low rates of oxygen consumption could be maintained if membranes of sufficiently high oxygen permeability are used. For primary hepatocytes, which have much higher oxygen demands, radial transport of oxygen is rate limiting, and the axial-flow hollow fiber cartridge is thus an inappropriate design for use as a bioartificial liver with primary hepatocytes. PMID- 10816202 TI - Development of the membrane autotransfusion system prototype-II: MATS-II. AB - This article is the second of a two-part series describing a membrane autotransfusion system, MATS, utilizing plasmapheresis technology. Based on experiences obtained from the first prototype (MATS-I), optimum blood filtration parameters with refined blood and flux pump synchronization were put into an original CPU-board and loaded on a miniaturized, self-operative, and preclinical prototype (MATS-II). This study was conducted to evaluate the MATS-II using diluted blood of various hematocrit concentrations. The results proved that this device could concentrate 4,000-10,000 ml of various hematocrit concentrations into higher than 40% while automatically controlling the flow speed from 250 to 400 ml/min. Also, no significant damage was generated to the red blood cells (RBC). Moreover, the MATS-II salvaged over 90% of platelets together with the RBC. These results suggest that the MATS-II achieves all clinical requirements of an autotransfusion device; it is a continuous hemoconcentration device with minimum damage to cellular components of the blood. PMID- 10816203 TI - The effect of sandblasting treatment on endurance properties of titanium alloy hip prostheses. AB - Sandblasting is a procedure of increasing surface roughness. This treatment is common in the orthopedic field. An increased roughness may affect the endurance limit of the material. This study investigates the effect on the endurance limit of the Ti6Al4V due to two different sandblasting treatments: fine sandblasting and coarse sandblasting. Twenty hip stems, 10 finely sandblasted and 10 coarsely sandblasted, were tested under sinusoidal fluctuating bending. The staircase method was used to estimate the endurance limit of the material. The results show an important reduction in the endurance properties up to 40% for the coarsely sandblasted specimens. The failures of the sandblasted specimens were not due to material defects. Rather, the decreased endurance strength of the sandblasted stems was caused by surface defects, which act as crack initiators. By modulating the roughness with an appropriate sandblasting treatment, it is possible to limit the reduction in the endurance limit of the alloy. PMID- 10816204 TI - Modified ultrafiltration improves carbon dioxide removal after cardiopulmonary bypass in infants. AB - Little is known about the role of modified ultrafiltration in ameliorating the adverse effects of the cardiopulmonary bypass on pulmonary function in infants. Twenty-nine nonrandomized consecutive infants (<12 months of age) who underwent unrestrictive ventricular septal defect closure between 1995 and 1998 were included in this study. Down's syndrome was associated in 9 patients. The actual ventilator settings were highly homogeneous among all patients and each time point in the study. Fourteen infants received modified ultrafiltration after the discontinuation of cardiopulmonary bypass. Fifteen untreated patients served as the control group. Correlates of cardiac and pulmonary functions for both groups were compared. The arterial carbon dioxide tension in the experimental group was significantly lower than in the control group from 20 to 240 min after bypass. Arterial oxygenation and pulmonary arterial pressure were similar in the 2 groups. Modified ultrafiltration improves carbon dioxide removal after cardiopulmonary bypass in infants. This may potentially convey a beneficial impact on hemodynamics. PMID- 10816205 TI - Continuous renal replacement therapy and charcoal plasmaperfusion in treatment of amanita mushroom poisoning. AB - Hemoperfusion has been used in the treatment of mushroom poisoning for many years. The aim of this study was to study the efficacy of charcoal plasmaperfusion (CPP) and continuous renal replacement therapy (CRRT) in 2 patients severely poisoned by the amanita mushroom. Both patients arrived at the ICU from another hospital with a diagnosis of amanita phalloides mushroom poisoning. The patients were precociously treated with CRRT for 20 h and CPP for 3 h every day. The treatments were effected for 3 and 5 days, respectively. Both patients recovered completely and were discharged asyntomatic after 7 and 10 days. PMID- 10816206 TI - Effect of the sinus of valsalva on the closing motion of bileaflet prosthetic heart valves. AB - Conventional bileaflet prosthetic mechanical heart valves close passively with backflow. Naturally, the valve has problems associated with closure, such as backflow, water hammer effect, and fracture of the leaflet. On the other hand, in the case of the natural aortic valve, the vortex flow in the sinus of Valsalva pushes the leaflet to close, and the valve starts the closing motion earlier than the prosthetic valve as the forward flow decelerates. This closing mechanism is thought to decrease backflow at valve closure. In this study, we propose a new bileaflet mechanical valve resembling a drawbridge in shape, and the prototype valve was designed so that the leaflet closes with the help of the vortex flow in the sinus. The test valve was made of aluminum alloy, and its closing motion was compared to that of the CarboMedics (CM) valve. Both valves were driven by a computer controlled hydraulic mock circulator and were photographed at 648 frames/s by a high speed charge-coupled device (CCD) camera. Each frame of the valve motion image was analyzed with a personal computer, and the opening angles were measured. The flow rate was set as 5.0 L/min. The system was pulsed with 70 bpm, and the systolic/diastolic ratio was 0.3. Glycerin water was used as the circulation fluid at room temperature, and polystyrene particles were used to visualize the streamline. The model of the sinus of Valsalva was made of transparent silicone rubber. As a result, high speed video analysis showed that the test valve started the closing motion 41 ms earlier than the CM valve, and streamline analysis showed that the test valve had a closing mechanism similar to the natural one with the effect of vortex flow. The structure of the test valve was thought to be effective for soft closure and could solve problems associated with closure. PMID- 10816207 TI - Analysis of hemodynamic response with 1/R control on biventricular bypass goat. AB - A conductance and arterial pressure based method (1/R control) to determine the cardiac output (CO) of a total artificial heart (TAH) was developed to provide a central nervous system with control over the output of TAH. In order to clarify the deference in hemodynamic response between natural heart and 1/R control, biventricular bypass was introduced in the goat. After 2 pneumatically driven sac type blood pumps were connected to the natural heart, the pulmonary artery was totally clamped to acquire 100% right heart bypass, and the ascending aorta was stenosed to acquire about 60 to 90% left heart bypass; 1/R control was performed substituting the output of the right artificial heart for the CO. The results demonstrated that stable control could be achieved. A discrepancy was often seen between the pulse rate (PR) of the artificial heart and the heart rate (HR) in absolute value. However, the relative changes of PR were quite similar to that of HR for the most part, indicating that the responses of 1/R control were the duplication of natural cardiac responses in normal daily activity. PMID- 10816208 TI - Estimation of pump flow rate and abnormal condition of implantable rotary blood pumps during long-term in vivo study. AB - The control system for an implantable rotary blood pump is not clearly defined. A detection system is considered to be necessary for pump flow monitoring and abnormal conditions such as back flow or a sucking phenomenon where the septum or left ventricle wall is sucked into the cannula, etc. The ultrasound flowmeter is durable and reliable but the control system should not be totally dependent on the flowmeter. If the flowmeter breaks, the rotary blood pumps have no control mechanism. Therefore, the authors suggest controlling the pumps by an intrinsic parameter. One left ventricular assist device (LVAD) calf model was studied where the flow rate and waveform of the pump flow proved to identify the sucking phenomenon. Thus, the pump flow rate was calculated from the required power, motor speed, and heart rate. The value of the coefficient of determination (R2) between the measured and estimated pump flow rate was 0.796. To estimate this abnormal phenomenon, 2 methods were evaluated. One method was the total pressure head in which the pump flow rate and motor speed were estimated. During normal conditions the total pressure head is 79.5 +/- 7.0 mm Hg whereas in the abnormal condition, it is 180.0 +/- 2.8 mm Hg. There was a statistical difference (p < 0.01). Another method is using a current waveform. There is an association between the current and pump flow waves. The current was differentiated and squared to calculate the power of the differentiated current. The normal range of this value was 0.025 +/- 0.029; the abnormal condition was 11.25 +/- 15.13. There was a statistical difference (p < 0.01). The predicted flow estimation method and a sucking detection method were available from intrinsic parameters of the pump and need no sensors. These 2 methods are simple, yet effective and reliable control methods for a rotary blood pump. PMID- 10816209 TI - Communications: burl osborne pioneer organ replacement hero award presentation PMID- 10816210 TI - The conservative management of acoustic neuroma: a review of forty-four patients with magnetic resonance imaging. PMID- 10816211 TI - Aetiological investigations of hearing loss in childhood: a review. PMID- 10816212 TI - The clinical value of septal perforation biopsy. AB - Biopsy from the edge of a septal perforation is performed to diagnose potentially significant aetiological conditions, including malignant pathology and Wegener's granulomatosis. However, it is common for the histological result to be undiagnostic and unhelpful in planning an individual patient's management. To clarify the role of septal perforation biopsy, we reviewed all biopsies performed on our patients over a 13-year period. Of the 65 biopsies performed, in 63 patients, none had a histological result which changed the clinical diagnosis made before biopsy. Forty-three patients had clinically benign perforations biopsied, and no histology other than chronic inflammation was obtained. In 16 patients with a clinical vasculitis, none had this diagnosis confirmed histologically. One positive biopsy was obtained from two patients with clinically malignant perforations yet both were treated for T-cell lymphoma. The subsequent management of all patients was based on their clinical diagnosis rather than their biopsy results. We suggest only clinically malignant perforations are worthwhile biopsying. PMID- 10816213 TI - Comparative audit of tonsillectomy. AB - The Comparative Audit Service of the Royal College of Surgeons of England undertook an audit of tonsillectomy throughout England and Wales in 1997. There were three parts to the audit: a management questionnaire related to tonsillectomy and its indications; an inpatient questionnaire for patients undergoing tonsillectomy; and a postoperative patient questionnaire. One hundred and fourteen ENT consultants returned the management questionnaire, 2450 questionnaires on inpatient details were returned by 132 consultants, and 1408 patients returned postoperative questionnaires. Two-thirds of the patients underwent tonsillectomy for recurrent acute tonsillitis, and most stayed 1 night in hospital; only 3% were day cases. Postoperative haemorrhage necessitated return to theatre for 0.75% of patients. Almost 7% of patients had to be readmitted after discharge. Overall, 92% of patients felt that their throat was better than before the operation, but slightly less than this number (88%) were pleased that they had the operation. The results are further discussed in the paper. The ability to cross-reference each part of the audit for individual patients proved extremely useful. PMID- 10816214 TI - A modified spacer device for inhalational drug therapy for chronic bronchitis/asthma in laryngectomised patients. AB - Following total laryngectomy, patients who need to continue their inhalational drug therapy for chronic pulmonary conditions experience a great deal of difficulty in administering the drugs. We describe a new spacer device to deliver inhalational drug therapy to laryngectomized patients which is simple to use and does not interfere with the patient's life style. The device involves a standard metered dose inhaler, a paediatric face mask and a small volume spacer. This device was used successfully in six laryngectomised patients for 12-18 months, who needed to continue their inhalational drug therapy for their pulmonary condition. PMID- 10816215 TI - Automated electrogustometry: a new paradigm for the estimation of taste detection thresholds. AB - Electrogustometery, first introduced in the 1950s for the clinical assessment of taste function, has returned to favour in recent years. This is due, in part, to the credence given to the procedure by an increased understanding of the mechanisms of taste transduction. The reliability and validity of electrogustometric assessments of taste function should increase with the use of automated testing. An automated computer-controlled testing paradigm was used to obtain threshold estimates simultaneously from the left and right sides of the tongue in 97 volunteers aged between 17 and 63 years. A small but statistically significant advantage was found for thresholds from the right side of the tongue (P<0.01). The range of thresholds, and of interside differences obtained with this procedure, are reported, and practical issues related to testing are discussed. PMID- 10816216 TI - The development of the stria vascularis in the human foetus. AB - The development of the stria vascularis in the human cochlea was studied in step sections of 81 human foetal temporal bones. The stria vascularis primordium can be identified as a ridge of epithelial cells on the lateral wall of the cochlear duct. The first signs of differentiation appear at the 11th week, but it is not until the 17th-18th week that the typical trilaminar structure is observed. The appearance of similar cells with notched nuclei in both marginal and mesenchymal layers at this stage suggests the possibility that some of the intermediate cells may be of epithelial origin. By the 21st week, the overall appearance resembles that of the adult structure. This occurs 1 week after the opening of the tunnel of Corti, and possibly marks the onset of cochlear function. PMID- 10816217 TI - Treatment of lymphangiomas of the head and neck in children by intralesional injection of OK-432 (Picibanil). AB - The treatments previously used for lymphangiomas of the head and neck in children surgery and intralesional injection of sclerosants-are associated with significant morbidity. A new treatment-intralesional injection of OK-432-was used for lymphangiomas of the head and neck in 11 children. The results were total shrinkage in two, marked shrinkage in two, slight shrinkage in five and no response in two. The results were not affected by previous surgery nor by whether aspiration prior to injection was possible. There were no recurrences in those children in whom shrinkage occurred and no child had subsequent surgery following injection. The results of this series support those of previous series showing that OK-432 injection is an effective and safe treatment for lymphangiomas of the head and neck in children. PMID- 10816218 TI - Parameters for the preoperative evaluation of arteriosclerosis for free-flap transfers in head and neck surgery. AB - The microsurgical transfer of free tissue has become essential for reconstructive surgery in the head and neck, and arteriosclerosis is one of the risk factors for microvascular anastomosis. In order to detect severe arteriosclerosis and to determine the parameters useful for evaluating arteriosclerosis preoperatively, the grade of arteriosclerosis was investigated with respect to age, PWV (Aortic Pulse Wave Velocity), ocular fundus findings, and histopathological findings of the recipient arteries in 40 patients with malignant tumours. Severe arteriosclerosis was detected in 2/40 patients by the PWV, in the same 2/40 patients by the ocular fundus findings and in the same 2/40 patients by histopathological findings. The current observations indicate that the PWV and ocular fundus findings are useful parameters for the preoperative evaluation of the grade of arteriosclerosis. PMID- 10816219 TI - Herpes zoster oticus treated with acyclovir and prednisolone: clinical manifestations and analysis of prognostic factors. AB - Herpes zoster oticus is a cranial polyneuropathy with facial nerve involvement as its main feature. The prognosis of the facial palsy is usually poor. Thirty patients with herpes zoster oticus suffering from facial palsy were admitted for parenteral acyclovir and oral prednisolone. Multiple regression analysis of improvement of facial palsy showed three significant covariates: age, multiple nerve palsies, and the initial grading of the palsy. The recovery of the facial palsy treated with acyclovir and prednisolone was good, and possibility of a good outcome was greater when the initial grade of the palsy was higher. Multiple nerve palsies and age had negative effects on the improvement. PMID- 10816220 TI - Sensorineural hearing loss in ulcerative colitis. AB - Sensorineural hearing loss has been described in a small number of patients with ulcerative colitis. At present, it is not known whether this is a rare and sporadic association, or whether these reports represent an under recognised extra-intestinal manifestation of ulcerative colitis. The aim of this study was to determine the prevalence of hearing loss in patients with ulcerative colitis. Twenty patients with active ulcerative colitis (mean age 45 years) were recruited prospectively along with 20 healthy age- and sex-matched controls. Otoscopy, tympanometry and pure tone audiometry were performed. The activity of the disease was scored according to a 12-month cumulative disease activity index. No patients complained of auditory symptoms. Otoscopy and tympanometry were normal in all patients and controls. Pure tone audiometry showed significant sensorineural hearing loss over all frequencies in patients with ulcerative colitis compared with controls. This preliminary study indicates that subclinical sensorineural hearing loss is associated with ulcerative colitis. Further study is warranted to determine the nature of the relationship and its pathological basis. PMID- 10816221 TI - Partial laryngectomy for recurrent laryngeal carcinoma. AB - From July 1975 to January 1998, 33 patients underwent partial laryngeal resection for residual or recurrent tumour after primary radical radiotherapy. Sixteen patients had T1 tumours on presentation, 14 were T2 and three were T3. Six patients underwent a supraglottic (horizontal) laryngectomy, 24 had a vertical partial laryngectomy, two had an endoscopic laser resection and one had an endoscopic laser resection followed by a vertical partial laryngectomy. The median time interval between radiotherapy and salvage surgery was 10 months (range 2-188 months). The median follow-up period was 41 months (range 12-185 months). There were five major postoperative complications (15%); two patients developed a pharyngeal fistula and three required further surgery for laryngo tracheal stenosis. Twenty-five patients (76%) retained their larynx with satisfactory speech and swallowing. Eight patients (24%) had to be converted to a total laryngectomy, seven for recurrent disease and one for laryngeal stenosis. Of the eight patents converted, seven had normal swallowing and six developed good tracheo-oesophageal speech. Seven patients (21%) developed recurrent tumour after partial laryngectomy and were subjected to total laryngectomy; six of these seven were salvaged. Only one of the 33 patients died with recurrent tumour, giving an ultimate disease-related survival of 97%. Conservation laryngeal surgery for salvage of selected patients who fail radical radiation therapy is safe, effective, and results in reasonable preservation of laryngeal function. PMID- 10816222 TI - Quality of life issues in recurrent respiratory papillomatosis. AB - Twenty-six adult patients attending the Royal National Throat, Nose, and Ear Hospital with Recurrent Respiratory Papillomatosis completed two postal questionnaires. One was the generic Short Form-36 (SF-36) quality of life instrument. Severely affected patients had lower scores in all dimensions of this instrument, with large differences from normal controls in dimensions of pain, physical limitation, and energy/vitality. The second questionnaire was newly devised, and designed to ask about a wide variety of larynx-specific symptoms. Answers by patients were compared with those of normal controls, and symptoms selected as significantly more likely to be reported by patients were studied further. Correlation was seen with clinical parameters, and questions likely to be responsive to clinical change in disease burden were identified. PMID- 10816223 TI - Children with recurrent episodes of acute otitis media: the effect of local administration of immunoglobulin G on acute otitis media, colonization and turnover of non-encapsulated Haemophilus influenzae in the nasopharynx. AB - In most children with recurrent episodes of acute otitis media (AOM), tube treatment is successful, but there are those who nevertheless suffer from middle ear infections. The aim of the present study was to ascertain whether local administration of immunoglobulin could reduce the number of episodes of otorrhoea in otitis-prone infants <2 years old who were treated with tubes, or whether it could affect the nasopharyngeal colonization and turnover of bacterial pathogens in the nasopharynx. IgG or placebo were also administered intranasally daily for 6 months to 50 infants, randomized in a double-blind study. An arbitrarily primed polymerase chain reaction (AP-PCR) was used to characterize the different isolates of NTHI (non-encapsulated, non-typable Haemophilus influenzae). Three infants in the IgG group and six infants in the control group suffered from > or =3 episodes of acute otitis media. No effect on the nasopharyngeal colonization or the turnover of non-encapsulated H. influenzae in the nasopharynx could be detected in either group. PMID- 10816224 TI - A history of chemical peeling. PMID- 10816225 TI - Transcatheter duplex ultrasound-guided sclerotherapy for treatment of greater saphenous vein reflux: preliminary report. AB - BACKGROUND: Surgical ligation and stripping of the greater saphenous vein has been the gold standard for treatment of saphenofemoral junction incompetence for several years. Although sclerotherapy of the greater saphenous vein has also been advocated by some phlebologists, the procedure can be technically challenging and has resulted in inadvertent nontarget injection. OBJECTIVE: The purpose of this study was to assess the effectiveness and safety of transcatheter duplex-guided sclerotherapy for the treatment of varicose veins due to saphenofemoral junction reflux. METHODS: Fifty-one greater saphenous veins in 50 patients were treated with transcatheter sclerotherapy. Using local anesthesia and ultrasound guidance, the greater saphenous vein was entered 15-45 cm below the saphenofemoral junction. An infusion catheter was placed over a guidewire and positioned under ultrasound guidance, and 3% sodium tetradecyl sulfate was administered below the saphenofemoral junction and along the course of an "empty" greater saphenous vein via the catheter. RESULTS: Catheter placement and treatment was possible in all patients, with 2-5 ml of 3% sodium tetradecyl sulfate administered per session. At the 24-hour and 1-week follow-ups, all treated greater saphenous vein segments were closed following initial treatment, with no flow detectable by continuous wave or color Doppler interrogation. No patients required re-treatment, with all veins remaining closed at 2- to 12-months follow-up. There have been no adverse reactions. CONCLUSION: Transcatheter duplex ultrasound-guided sclerotherapy should improve both the safety and efficacy of treatment compared to conventional ultrasound-guided sclerotherapy and offers an alternative to surgical ligation and stripping for those patients wishing to avoid surgery. PMID- 10816226 TI - Ber-EP4-positive phenotype differentiates actinic keratosis from superficial basal cell carcinoma. AB - BACKGROUND: Well-defined histopathologic criteria exist to distinguish actinic keratosis (AK) from superficial basal cell carcinoma (BCC). A similar morphology of downwardly budding dysplastic keratinocytes may occur in both entities, creating potential for errors in diagnosis. A marker that could reliably distinguish these two lesions would overcome this difficulty in diagnosis. OBJECTIVE: To investigate whether Ber-EP4 staining is useful in distinguishing AK from superficial BCC, and to determine whether AK exhibits a cellular phenotype that is more consistent with BCC or squamous cell carcinoma (SCC). METHODS: We subjected tissue sections from superficial BCC, AK, and squamous intraepithelial neoplasia (SIN) demonstrating epidermal budding to immunohistochemical staining with monoclonal antibody Ber-EP4. RESULTS: Abnormal keratinocytes in all specimens of superficial BCC (5 of 5) were Ber-EP4 positive; all AK (10 of 10) and SIN (8 of 8) were Ber-EP4 negative. CONCLUSION: Ber-EP4 staining reliably distinguishes AK from superficial BCC. The lack of Ber-EP4 staining of AK supports the currently accepted pathogenetic dogma that SIN and SCC arise from AK, but BCC does not. PMID- 10816227 TI - Laser hair transplantation II. AB - BACKGROUND: Micrografting has revolutionized hair transplantation, resulting in far more natural results than have been achieved prior to this technique. Operative problems including bleeding, graft compression, and ease of graft insertion have been alleged to be decreased with the use of a high-energy pulsed CO2 laser for creation of recipient sites. However, because of thermal injury to surrounding tissue, diminished hair growth and slower wound healing also have been seen. OBJECTIVE: A new laser handpiece, designed to minimize tissue thermal damage to hair recipient sites was tested in comparison to 18-gauge needle recipient sites in the same patients. Hair growth counts at 6 months as well as various measurements of operative problems were compared. METHODS: Two hundred laser grafts were performed on one scalp side and compared to 200 needle grafts on the opposite side. Hair counts preoperatively and at 6 months were performed. Biopsies for depth and surrounding tissue damage were taken intraoperatively. Time for completion of various stages of the procedure for each side were recorded, as well as the incidence of side effects per side. RESULTS: Hair growth was equal for each side. Bleeding and operative time were significantly reduced and ease of graft insertion was increased on the laser side. Graft compression and scarring were not seen. CONCLUSIONS: Laser hair transplantation is faster than using conventional techniques and results in equal hair growth, and should be considered as a viable alternative technique. PMID- 10816228 TI - A single postoperative application of nitroglycerin ointment does not increase survival of cutaneous flaps and grafts. AB - BACKGROUND: Nitroglycerin is a vasodilator that has been reported to improve cutaneous flap and graft survival. It has not been tested in controlled studies. OBJECTIVE: We designed our study to test the effectiveness of a single postoperative application of nitroglycerin on flap and graft survival. METHODS: Eighty-eight surgical repairs received topical nitroglycerin and 85 received control ointment (polysporin). Treatment sites were evaluated on postoperative day 7 and assigned a percentage of surface area survival. RESULTS: There was no significant difference in the complication rate of flaps and grafts treated with nitroglycerin (12.5%) compared with those treated with control ointment (8.4%) (P = .244). Subset analysis of flaps as a group and grafts as a group were not meaningful because the complication rates were so low. CONCLUSION: There is no survival increase of flaps and grafts treated with a single application of nitroglycerin ointment. PMID- 10816229 TI - Prospective study of hair reduction by diode laser (800 nm) with long-term follow up. AB - BACKGROUND: Multiple laser systems are available for the purpose of hair removal. OBJECTIVE: The purpose of this study was to determine the safety and long-term efficacy of the 800 nm, pulsed diode laser at reducing hair count. METHODS: Fifty volunteers, primarily Fitzpatrick skin types II and III, with dark brown or black hair, were treated with a diode laser (800 nm, 10-40 J/cm2, 5-30 msec, 9 mm 9 mm, 5 degrees C chilled handpiece). Each subject had eight treatment sites at varying fluences and pulse durations, as well as a varying number of treatments and pulses. Hair counts were obtained at each site at baseline, 1, 3, 6, 9, and an average of 20 months after treatment. RESULTS: After one treatment, hair regrowths ranged from 22 to 31% at the 1-month follow-up visit, then remained stable between 65 and 75% from the 3-month to the averaged 20-month follow-up. After two treatments there were relatively longer growth delays, with hair regrowths plateauing beginning at 6 months after treatment and ranging from 47 to 66% for the remainder of the follow-up evaluations. Side effects were limited to pigmentary changes, transient in subjects with skin types II and III. CONCLUSIONS: This 800 nm diode laser with a chilled sapphire tip and variable pulse duration is safe and effective for long-term hair reduction in individuals with skin types II and III. PMID- 10816230 TI - Effect of external ultrasound postliposuction: a side-to-side comparison study. AB - BACKGROUND: External ultrasound (EUS) has been used as a therapeutic modality for more than 30 years by multiple medical specialists, primarily for stimulating the repair of soft tissue injuries and relieving pain. It has also been used to treat postoperative swelling after liposuction. There are no controlled studies documenting the objective and subjective effects of EUS following liposuction. OBJECTIVE: The purpose of this preliminary study was to evaluate the role of EUS in the relief of postoperative symptoms following traditional tumescent liposuction. METHODS: Twenty-five patients underwent tumescent liposuction of the abdomen or hips. Within 2 weeks after the procedure, a double-blind study was initiated in which EUS therapy was applied at therapeutic energies to one-half of the treated area, while the other side was treated with placebo settings. Patients were treated biweekly for a total of six to eight treatments. Both objective and subjective parameters were assessed during the subsequent side-by side evaluations by the patient and observer. RESULTS: After four treatments, 18 of 25 patients had the same postoperative symptoms on each side. Two had fewer symptoms on the EUS side and five were worse on the EUS side. After completion of all treatments, 17 of 25 again had the same symptoms on the therapeutic and placebo sides. Three had improved symptoms and five were worse on the EUS side. The subjective findings revealed that all patients believed the treatment helped significantly on both the placebo and EUS-treated sides. CONCLUSION: EUS after liposuction of the abdomen at the study parameters provided no objective benefit compared to placebo in postoperative recovery. Patient acceptance and perception of benefit are high. Further studies are needed to determine the optimal timing and energies required for therapeutic value. PMID- 10816231 TI - Er:YAG laser for the treatment of actinic keratoses. AB - BACKGROUND: There is no single optimal treatment for multiple facial actinic keratoses. The existing therapies such as topical 5-fluorouracil, chemical peels, cryotherapy, dermabrasion, and CO2 laser resurfacing can produce prolonged recovery time or are often operator dependent. OBJECTIVE: The purpose of this study was to investigate another therapeutic modality which provides a shorter recovery time with uniform results. We performed a prospective pilot study investigating the use of the Er:YAG laser for the treatment of multiple facial actinic keratoses. METHODS: Five patients with multiple facial actinic keratoses were treated with two to three passes of Er:YAG laser. Anesthesia was achieved in all cases by topical application and local infiltration when indicated. All patients were treated with 2.0 J, 5 mm spot size, and a fluence of 10 J/cm2. Clinical and histologic evaluations were performed both pre- and postoperatively. RESULTS: All patients showed a decrease in the total number of clinical actinic keratoses on the face ranging from 86 to 96%. In addition to the reversal of actinic damage in the epidermis, histologic evidence revealed increased fibroplasia and decreased superficial solar elastosis 3 months after the laser resurfacing. Reepithelialization occurred in 5-8 days, and erythema lasted for about 3-6 weeks after the procedure. There was no evidence of scarring or pigmentary changes in any of the patients during the follow-up period. CONCLUSION: Er:YAG laser skin resurfacing is a safe and effective treatment for multiple facial actinic keratoses. Histologic data suggest a new zone of collagen deposition occurs in the superficial papillary dermis. Under our current parameters, Er:YAG laser skin resurfacing has a relatively short recovery period and a low risk of scarring. Unlike the CO2 laser, Er:YAG laser skin resurfacing can be performed with topical anesthesia alone. PMID- 10816232 TI - Injectability and tissue compatibility of poly-(N-vinyl-2-pyrrolidone) in the skin of rats: a pilot study. AB - BACKGROUND: Filling substances have been used in dermatologic surgery for decades, but an ideal agent has yet to be discovered. Poly-(N-vinyl-2 pyrrolidone) is a hydrogel that has been used in medical settings for more than 50 years, but not as a cutaneous filling agent. OBJECTIVE: We investigated the intracutaneous injectability and tissue compatibility of this hydrogel in a rat model. Particular attention was paid to ease of injection through small needles, volume retention of the implant, clinical course, and histocompatibility. METHODS: The shaved backs of 12 anesthetized Sprague-Dawley rats were injected with the sterilized hydrogel and the rats closely observed. The rats were sacrificed in groups of four at 2, 4, and 12 weeks after implantation. Implant size was measured, volume calculated, and biopsies taken at each time interval. RESULTS: Poly-(N-vinyl-2-pyrrolidone) is easily injected through 30-gauge needles. All rats tolerated the implants well clinically. Histopathology revealed well-circumscribed implants with pseudoencapsulation, neoangiogenesis, and mixed inflammatory cells predominating at the periphery. Volume calculations revealed an average of 33% reduction at 4 weeks and 35% reduction at 12 weeks. CONCLUSION: Poly-(N-vinyl-2-pyrrolidone) is easy to inject intracutaneously and is well tolerated in the rat model. Short-term volume retention is good. Histopathology suggests a subclinical inflammatory reaction expected with implantation of a synthetic substance into the skin. Additional studies are necessary to investigate the continued persistence of the hydrogel and its long-term effects on surrounding tissue. PMID- 10816233 TI - Level of fibronectin mRNA is markedly increased in human chronic wounds. AB - BACKGROUND: Acute wound healing has been extensively investigated over the years, however, little is known about possible healing defects in chronic wounds. Fibronectin (FN) plays a critical role in different phases of wound healing and has been demonstrated to be degraded in chronic wounds by proteases. Fibroblasts cultured from chronic leg ulcers showed a higher level of FN compared to normal fibroblasts. OBJECTIVE: We explored whether the increase in FN protein in chronic wounds is due to increased FN mRNA. In addition, the level of alpha5beta1 integrin FN cell surface receptor was also examined. METHOD: Skin biopsies were taken from normal skin within a few hours of Mohs surgery and from the edge of chronic venous leg ulcers. In situ hybridization was performed to determine the level of FN mRNA. The level of integrin alpha5beta1 was determined by immunohistochemistry. RESULTS: The level of FN mRNA in normal skin and acute wounds was undetectable. In contrast, FN mRNA was heavily induced throughout the dermis of chronic wounds. Immunostaining using a monoclonal antibody against the alpha5 subunit of integrin revealed that chronic wounds and normal skin showed undetectable levels of alpha5beta1 integrin. A large induction of alpha5 was observed in acute wounds. CONCLUSION: For reepithelization to occur, epidermal keratinocytes need to migrate over the wound surface, a process requiring an interaction between FN and its cell surface receptor integrin alpha5beta1. These findings suggest that although FN mRNA is increased in chronic wounds, lack of FN cell surface receptor may prevent migration of epidermal keratinocytes in chronic wounds. PMID- 10816234 TI - Closure of the greater saphenous vein with endoluminal radiofrequency thermal heating of the vein wall in combination with ambulatory phlebectomy: preliminary 6-month follow-up. AB - BACKGROUND: Incompetence of the saphenofemoral junction with reflux into the greater saphenous vein is one cause of chronic venous hypertension which may lead to the development of varicose and telangiectatic leg veins. Therefore treatment is necessary. OBJECTIVE: To evaluate a novel method for closing the incompetent greater saphenous vein at its junction with the femoral vein through an endoluminal approach. METHODS: Ten patients with reflux at the saphenofemoral junction into the greater saphenous vein were treated with radiofrequency heating of the vein wall through an endoluminal catheter. Patients were evaluated at 3 and 6 months to determine treatment efficacy as well as adverse sequelae. RESULTS: All treated patients achieved complete closure of the saphenofemoral junction and greater saphenous vein. Complete treatment took an average of 20 minutes. Adverse sequelae were minimal, with 2 of 12 patients having mild erythema for 2-3 days. CONCLUSION: Endoluminal radiofrequency thermal heating of an incompetent greater saphenous vein has been shown to be easily accomplished and efficacious throughout the 6-month follow-up period. PMID- 10816235 TI - Clinical comparative study between cryotherapy and local dermabrasion for the treatment of solar lentigo on the back of the hands. AB - BACKGROUND: Solar lentigo is a common and unsightly dermatosis that has a variety of proposed treatments. OBJECTIVE: This study was done to assess the efficacy and the effectiveness of localized dermabrasion compared with cryotherapy with liquid nitrogen on solar lentigo on the back of the hands. METHODS: Ten female patients aged 64-96 years with solar lentigo on the back of the hands were treated with dermabrasion or cryotherapy and observed over a 6-month period. RESULTS: The postsurgery signs and symptoms were less intense and better tolerated with localized dermabrasion. More than 50% of the patients treated with cryotherapy still had hypochromia in the treated areas 6 months after treatment, compared with 11% of the patients treated with dermabrasion. The percentage of recurrence was the same with the both treatments (55.55%). CONCLUSION: Localized dermabrasion is an efficacious and effective technique comparable to cryotherapy for the treatment of solar lentigo on the back of the hands. PMID- 10816236 TI - Involvement of keratinocyte activation phase in cutaneous graft healing: comparison of full-thickness and split-thickness skin grafts. AB - BACKGROUND: Little is known about keratinocytic activation in the graft take and healing process. OBJECTIVE: To investigate the clinical and molecular differences between pure epidermal sheet graft (PESG), split-thickness skin graft (STSG), and full-thickness skin graft (FTSG). METHODS: Three different thickness skin grafts (PESG, STSG, and FTSG) were performed onto three kinds of porcine wounds: shallow, deep, and full. Graft take, contraction, and Ki-67 and beta1 integrin expression in epidermis were studied. RESULTS: All grafts took well. As expected, full wounds covered by PESG and STSG contracted more than those covered by FTSG, whereas shallow wounds covered by FTSG contracted more than those covered by STSG. No difference in contracture was observed among deep wounds covered by PESG, STSG, and FTSG. Up-regulation of Ki-67 and beta1 integrin expression was greater in PESG and STSG, compared with little expression in FTSG. CONCLUSION: The keratinocytic activation phase may occur both in the STSG and PESG healing process, as well as serum imbibition, inosculatory, and revascularization phases. PMID- 10816237 TI - Lip rejuvenation. AB - BACKGROUND: The management of the senile lip remains a dilemma. Allogenic fillers often feel unnatural, lip resurfacing is not adequate, and fat transfers may disappear. OBJECTIVE: To develop a more reliable step-by-step approach to lip augmentation. METHOD: Lips were divided into types: (1) the simple senile lip that had lost its fullness-treated with fat augmentation; (2) the lip with rhagades-treated with fat augmentation and laser resurfacing; and (3) the duckbill lip-treated with lip advancement, fat augmentation, and laser resurfacing. Tattooing to increase the degree of redness was performed on all types of lips. RESULT: If there had been a previously full lip, it was possible to rejuvenate the lip by simple augmentation with autologous fat transfer. The average number of fat transfer sessions to generate this pleasing lip was two to three. The lip with rhagades required fat filling, with laser resurfacing to achieve a new contour. The duckbill lip required a lip advancement along with lipofilling and laser resurfacing. All types benefited from lip tattooing. CONCLUSIONS: It was possible with fat augmentation and/or laser resurfacing to generate a pleasing lip in type 1 and 2 lips. Lip type 3 required a lip advancement along with fat augmentation and laser resurfacing. Lip tattooing accentuated all the lip types. PMID- 10816238 TI - Treatment of Hori's nevus with the Q-switched Nd:YAG laser. AB - BACKGROUND: Hori's nevus is an acquired pigmented lesion involving bilateral blue brown facial macules. There has been a dearth of reported treatment modalities for this condition. OBJECTIVE: The purpose of this study was to evaluate the efficacy of the Q-switched Nd:YAG laser for the treatment of Hori's nevus. METHODS: The Q-switched Nd:YAG laser was used to treat Hori's nevus in 66 Asian patients. Patients were treated up to seven times. The follow-up time after the final treatment ranged from 3 to 44 months. RESULTS: Twenty six percent of patients showed good to excellent clearing after one to two treatments. Fifty percent of patients who underwent more than two treatments received good to excellent results. CONCLUSION: The Q-switched Nd:YAG laser can be used to treat Hori's nevus. Results are not as good as those seen with nevus of Ota. PMID- 10816239 TI - Instruction of compression therapy by means of interface pressure measurement. AB - BACKGROUND: Compression therapy of the leg is the cornerstone in the conservative treatment of venous ulcers. The application of compression bandages, however, is largely a matter of personal experience. OBJECTIVE: To evaluate the interface pressure under compression bandages and to improve the technique. METHODS: Six courses on wound healing with 24-28 participants as well as individual training at our hospital were provided. Interface pressure at the distal medial calf was measured using a simple, but accurate pressure sensor that was built for this purpose (accuracy: +/-3 mm Hg). RESULTS: During the wound healing courses, the absolute difference from the target pressure of 35-45 mmHg improved from 8.4 mm Hg (95% CI 0.0-34.1) to 3.5 mm Hg (95% CI 0.0-14.0) (P = .0001). After four sessions, interface pressures greater than 60 mmHg were avoided. During individual training, even nurses with everyday experience in compression therapy improved their accuracy. CONCLUSION: There is a need for objective measurement of interface pressure in the teaching of compression therapy with bandages. The principles can be taught during a few exercises. However, repeated practice over a longer period of time is necessary to reach a certain accuracy. PMID- 10816240 TI - The youthful forehead: placement of skin incisions in hidden furrows. AB - BACKGROUND: Attaining good aesthetic results remains a primary goal in removal of benign cutaneous facial lesions. OBJECTIVE: Strategic planning of the incision is perhaps the most critical step in excision of such a lesion. METHODS: A study of one case of epidermoid cyst excision from a youthful forehead was undertaken. RESULTS: Poor surgical planning of a simple cyst excision from the forehead resulted in placement of the incision inferior to a natural furrow and within the basin defect, producing a noticeable scar. Facial animation accentuates the aesthetically poor placement of the surgical incision. CONCLUSION: The detection of hidden furrows through facial animation during preoperative planning, especially in the youthful forehead, is imperative for achievement of an optimal aesthetic result. When possible, incisions should be concealed within natural furrows. PMID- 10816241 TI - Angiomatoid fibrous histiocytoma: a case report and review of the literature. AB - We describe the case of a 5-year-old boy found to have an enlarging mass of the posterior neck. The patient underwent an uneventful excision of the mass, and pathologic examination revealed it to be an angiomatoid fibrous histiocytoma. This case report presents a review of the recent literature of this rare soft tissue tumor. PMID- 10816242 TI - Use of disposable otoscope specula in cryotherapy. PMID- 10816243 TI - Stellate purse-string closure. AB - BACKGROUND: There are fundamental concepts we use in managing surgical defects. Whether planning a primary closure or a local flap, we frequently modify the basic design to maximize aesthetic outcomes, taking into consideration a number of factors including the location of the defect and tissue availability. OBJECTIVE: We describe the stellate modified purse-string closure, a novel flap modification. METHOD: Report of an illustrated case. RESULT: A patient with vertex scalp defect was reconstructed using the stellate purse-string flap. CONCLUSION: Certain modifications of commonly used reconstructive techniques can be utilized in specific situations to enhance cosmesis. Advantages of this modification are discussed. PMID- 10816244 TI - Modified Burow's wedge flap for upper lateral lip defects. AB - BACKGROUND: There are fundamental concepts we use in managing surgical defects. Whether planning a primary closure or a local flap, we frequently modify the basic design to maximize aesthetic outcomes, taking into consideration a number of factors including the location of the defect and tissue availability. OBJECTIVE: We describe a modified Burow's wedge flap for upper lateral lip defects. METHOD: Report of an illustrated case. RESULT: A patient with an upper lip defect was successfully reconstructed using the modified Burow's wedge flap, where the Burrow's wedge is placed on the mucocutaneous lip. CONCLUSION: Certain modifications of commonly used reconstructive techniques can be utilized in specific situations to enhance cosmesis. For the Burow's wedge flap, the dermatologic surgeon has several options in placing the Burow's triangle. This is an example of how alternatives in a closure can be used depending on the laxity of the skin and the size of the defect. Advantages and disadvantages of this alternative placement of the Burow's triangle are discussed. PMID- 10816245 TI - Artifacts in frozen section preparation. PMID- 10816246 TI - What is basosquamous carcinoma? PMID- 10816247 TI - A novel method to remove Monsel's stain. PMID- 10816248 TI - Signal to forces: central themes in cytokinesis. PMID- 10816249 TI - Multiple types of calcium signals are associated with cell division in zebrafish embryo. AB - Recent studies suggested that a Ca(2+) signal is involved in the regulation of cell division. For example, using a confocal imaging technique, we have shown that a localized Ca(2+) elevation was clearly associated with the onset of cytokinesis in zebrafish embryo [Chang and Meng (1995) J. Cell Biol. 131:1539 1545]. This finding was later confirmed in studies using aequorin as a Ca(2+) probe. Here, we used a 4-D confocal measurement technique to further characterize the properties of the Ca(2+) signal associated with cell division. We found evidence that there were three types of Ca(2+) signals associated with different stages of cell cleavage in embryonic cell. The first type was repetitive Ca(2+) spikes that emerged several minutes before the first cell cleavage began. These Ca(2+) spikes were first distributed broadly over the central region of the blastodisc and then gradually localized in the equatorial region; they appeared to play the role of determining the position of the first cleavage plane. The second type was a calcium wave that propagated along the cleavage furrow and appeared to guide the furrow extension during the progression of cytokinesis. The third type was a group of post-cleavage calcium spikes that appeared to be responsible for furrow deepening and maintenance of the contractile band. When this type of Ca(2+) transient was blocked by injecting BAPTA or heparin, cell cleavage regressed and the structure of the contractile band could no longer be maintained. PMID- 10816250 TI - Citron, a Rho target that affects contractility during cytokinesis. AB - The small GTPase Rho, which regulates cell shape, is thought to contribute to cytokinesis. Recently, Citron was characterized as a Rho target. This large protein contains a Ser/Thr kinase domain related to that of ROCK, another Rho effector. Both endogenous Citron and recombinant Citron localize to the cleavage furrow in dividing cells and to the midbody in post-mitotic cells. Moreover, overexpression of Citron deleted from its C-terminal sequence caused abnormal contractions specifically during cytokinesis, resulting in the formation of multinucleated cells. Cell shape, F-actin, intermediate filaments, and microtubules appeared essentially normal in these cells during interphase. Thus, Citron is a Rho effector that appears to function during cytokinesis, modulating its contractile process. In brain, however, Citron is highly expressed in a subset of neurons as a brain-specific isoform that lacks a kinase domain, Citron N. This protein accumulates in synapses and associates to the NMDA receptor via interaction with the adaptor protein PSD95, suggesting that the function of Citron is specialized in the neurons. PMID- 10816252 TI - Molecular biological approaches to study myosin functions in cytokinesis of Dictyostelium. AB - The cellular slime mold Dictyostelium discoideum is amenable to biochemical, cell biological, and molecular genetic analyses, and offers a unique opportunity for multifaceted approaches to dissect the mechanism of cytokinesis. One of the important questions that are currently under investigation using Dictyostelium is to understand how cleavage furrows or contractile rings are assembled in the equatorial region. Contractile rings consist of a number of components including parallel filaments of actin and myosin II. Phenotypic analyses and in vivo localization studies of cells expressing mutant myosin IIs have demonstrated that myosin II's transport to and localization at the equatorial region does not require regulation by phosphorylation of myosin II, specific amino acid sequences of myosin II, or the motor activity of myosin II. Rather, the transport appears to depend on a myosin II-independent flow of cortical cytoskeleton. What drives the flow of cortical cytoskeleton is still elusive. However, a growing number of mutants that affect assembly of contractile rings have been accumulated. Analyses of these mutations, identification of more cytokinesis-specific genes, and information deriving from other experimental systems, should allow us to understand the mechanism of contractile ring formation and other aspects of cytokinesis. PMID- 10816251 TI - Cytokinesis in Tetrahymena: determination of division plane and organization of contractile ring. AB - A protein, Tetrahymena p85, is localized to the presumptive division plane before the formation of the contractile ring. p85 directly interacts with Tetrahymena calmodulin (CaM) in a Ca(2+)-dependent manner, and p85 and CaM colocalize in the division furrow. A Ca(2+)/CaM inhibitor N-(6-Aminohexyl)-5-chloro-1 naphthalenesulfonamide HCl (W7) inhibits the direct interaction between p85 and Ca(2+)/CaM. W7 also inhibits the localization of p85 and CaM to the division plane, and the formation of the contractile ring and division furrow. Tetrahymena fimbrin and elongation factor-1a (EF-1alpha), which induce bundling of Tetrahymena F-actin, are also localized to the division furrow during cytokinesis. The Tetrahymena fimbrin has two actin-binding domains, but lacks the EF-hand Ca(2+)-binding motif, suggesting that Tetrahymena fimbrin probably cross links actin filaments in a Ca(2+)- insensitive manner during cytokinesis. The evidence also indicates that Ca(2+)/CaM inhibits the F-actin-bundling activity of EF-1alpha; and EF-1alpha and CaM colocalize in the division furrow. In this review, we propose that the Ca(2+)/CaM signal and its target protein p85 cooperatively regulate the determination of the division plane, and that a Ca(2+) insensitive actin-bundling protein, Tetrahymena fimbrin, and a Ca(2+)/CaM sensitive actin-bundling protein, EF-1alpha, play pivotal roles in regulating the organization of the contractile ring microfilaments. PMID- 10816253 TI - Molecular analysis of racE function in Dictyostelium. AB - Dictyostelium has long proven to be a valuable system for studying various aspects of the cytoskeleton and cell motility. In this review we describe the isolation of a novel gene, racE, and how we have used multiple approaches to learn how the product of this gene is involved in cytokinesis. The racEgene was isolated in a screen designed to identify genes specifically required for cytokinesis. The use of GFP fusion proteins, coupled with mutational analysis, allowed us to determine that racE exerts its function at the plasma membrane throughout the entire cell cycle. Measurements of cortical tension and observations of live cells in suspension culture revealed that racE is involved in the regulation of cortical tension and actin organization at the cortex. We postulate that in the absence of proper cortical tension, cytokinesis cannot proceed normally. PMID- 10816254 TI - Cytokinesis in fission yeast: a myosin pas de deux. AB - Cytokinesis in the fission yeast, Schizosaccharomyces pombe consists of two distinct but overlapping events: the assembly and constriction of a cytokinetic actomyosin ring (CAR) and the formation of a cross wall or septum. These two processes must be spatially and temporally coordinated both with each other and with other cell cycle events, most notably spindle formation and anaphase chromosome segregation. In fission yeast, the CAR contains two unusual type II myosins, Myo2, encoded by the gene myo2(+), and Myp2, encoded by myp2(+). The relationship of these two proteins to each other and their relative contribution to CAR assembly and contraction is largely unknown. Here we review what is known about the role of each myosin in cytokinesis and present some new information concerning their regulation and possible physical interaction. PMID- 10816255 TI - Microtubule and actin-dependent movement of the formin cdc12p in fission yeast. AB - Although a number of gene products involved in cytokinesis have been identified, still little is known about how these proteins are localized to the proper site and assembled into a ring structure. How is the plane of cell division is positioned in the cell? Schizosaccharomyces pombe are simple rod-shaped eukaryotic cells that divide by medial fission using a medial contractile ring. S. pombe cdc12p encodes a member of the formin gene family, proteins with conserved roles in cytokinesis and actin organization. cdc12p is required specifically for the formation of the medial ring and is located in this ring during mitosis. Time-lapse microscopy of cells expressing GFP-cdc12p protein fusions reveals that during interphase, S. pombe cdc12p is present in a discrete, motile cytoplasmic particle that moves using both actin and microtubules. At the onset of mitosis, the spot moves to the future site of cell division and spreads out into a ring. These studies demonstrate that a cytokinesis factor may travel on both microtubule and actin networks to the site of contractile ring assembly. These findings suggest a potential mechanism for how the mitotic spindle positions the cell division plane in animal cells. PMID- 10816256 TI - Involvement of PCH family proteins in cytokinesis and actin distribution. AB - Pombe Cdc15 homology (PCH) proteins constitute an extensive protein family whose members have been found in diverse eukaryotic organisms. These proteins are characterized by the presence of several conserved sequence and structural motifs. Recent studies in yeast and mammalian cultured cells have implicated these proteins in actin-based processes, in particular, cytokinesis. Here we review the recent findings on the in vivo localization, function, and binding partners of PCH family members. We also provide new microscopy data regarding the in vivo dynamics of a budding yeast PCH protein involved in cytokinesis. PMID- 10816257 TI - Regulation of intermediate filament organization during cytokinesis: possible roles of Rho-associated kinase. AB - Intermediate filaments (IFs), which form the structural framework of cytoskeleton, have been found to be dramatically reorganized during mitosis. Some protein kinases activated in mitosis are thought to control spatial and temporal IF reorganization through phosphorylation of IF proteins. Rho-associated kinase (Rho-kinase), one of the putative targets of the small GTPase Rho, does phosphorylate IF proteins, specifically at the cleavage furrow during cytokinesis. This cleavage furrow-specific phosphorylation plays an important role in the local IF breakdown and efficient separation of IF networks. Recent studies on Rho signaling pathways have introduced new models about the molecular mechanism of rearrangements of cytoskeletons including IFs during cytokinesis. PMID- 10816258 TI - Real-time high-resolution optical sectioning suggests biphasic cytokinetic mechanism in Dictyostelium discoideum. AB - Despite its biological significance, much of the mechanism of cytokinesis is not yet resolved. The problems include: (1) signaling mechanism determining the position of the cleavage furrow, (2) molecular and mechanistic nature of the contractile ring, and (3) the origin of forces responsible for cleavage. Using high-resolution imaging technique, the present study analyzes morphometric changes of cytokinesis in wild type (NC4) Dictyostelium discoideum amoeba. A sample was prepared by the agar-overlay method, creating 3-mm-thick, nearly two dimensional cells; and high-resolution image was acquired at 16.7 milliseconds' temporal, 234 nm x, y-, and 100 nm z-axis resolutions. Under this condition, the formation of cleavage furrow initiates at mitotic telophase, and daughter cells separate 18-22 minutes after the furrow initiation. We found that the compression of cells and the room temperature need to be carefully controlled for cytokinesis to proceed in an orderly manner. The results demonstrate that the pole-to-pole distance increases by 83% during the initial 5 minutes of cytokinesis, while the distance of equator only decreases by 56%. In contrast, during the subsequent 5 minutes, the pole-to-pole distance only increases by 17%, while the equator distance decreases as much as by 44%. This study indicates that cytokinesis consists of at least two different phases, each of which results from a different mechanism. PMID- 10816259 TI - Assembly of cytoskeletal proteins into cleavage furrows of tissue culture cells. AB - We review results obtained after fluorescent actin and myosin II probes were microinjected into interphase and prophase PtK2 and LLC-PK tissue culture cells to follow the changing distribution of these cytoskeletal proteins in the live cells during division. The fluorescent probes first begin to assemble into the future furrow region during mid-anaphase before any sign of initial contractions. The total concentrations of F-actin and myosin in the cleavage furrow begin to decrease a few minutes after the onset of furrow contraction. The cell's shape and the position of its mitotic spindle affect the deposition of cytoskeletal proteins in the forming cleavage furrow. In cells with two spindles, contractile proteins were recruited not only to the cortex bordering the former metaphase plates but also to the cortex midway between each pair of adjacent non-daughter poles or centrosomes. The furrowing between adjacent poles seen in these cultured cells are similar to the furrows observed by Rappaport [(1961) J Exp Zool 148:81 89] when echinoderm eggs were manipulated into a torus shape so that the poles of two mitotic spindles were adjacent to one another. These observations on injected tissue culture cells suggest that vertebrate cells share common mechanisms for the establishment of the cleavage furrow with echinoderm cells. PMID- 10816260 TI - Relationships between the central spindle and the contractile ring during cytokinesis in animal cells. AB - During late anaphase and telophase, animal cells develop a bundle of antiparallel, interdigitating microtubules between the two daughter nuclei. Recent data indicate that this structure, called the central spindle, plays an essential role during cytokinesis. Studies in Drosophila and on vertebrate cells strongly suggest that the molecular signals for cytokinesis specifically emanate from the central spindle midzone. Moreover, the analysis of Drosophila mutants defective in cytokinesis has revealed a cooperative interaction between the central spindle microtubules and the contractile ring: when either of these structures is perturbed, the proper assembly of the other is disrupted. Based on these results we propose a model for the role of the central spindle during cytokinesis. We suggest that the interaction between central spindle microtubules and cortical actin filaments leads to two early events crucial for cytokinesis: the positioning of the contractile ring, and the stabilization of the plus ends of the interdigitating microtubules that comprise the central spindle. The latter event would provide the cell with a specialized microtubule scaffold that could mediate the translocation of plus-end-directed molecular motors to the cell's equator. Among the cargoes transported by these motors could be proteins involved in the regulation and execution of cytokinesis. PMID- 10816261 TI - New technique for examining invertebrate membranes using high resolution scanning electron microscopy. PMID- 10816262 TI - Evaluation of fixation methods for ultrastructural study of Caenorhabditis elegans embryos. AB - Various fixation methods for transmission electron microscopy (TEM) were tested on Caenorhabditis elegans embryos. By combining various techniques, using 3.4% chitinase in combination with 1% alpha-chymotripsin, we were able to establish a new fixation procedure that for the first time preserves both membranes and internal cellular ultrastructure of C. elegans embryos in different stages of development. This unique procedure will enable a hitherto unattainable standard for TEM research of C. elegans embryos. Sectioning of specific developmental stages fixed with this method allows a detailed study of ultrastructural aspects of embryogenesis. PMID- 10816263 TI - Ins and outs of digital electron microscopy. AB - In order to do any sort of computer processing of the images produced by an electron microscope they have to be digitized. Therefore, any form of microscopy that involves computer processing has to be considered as digital microscopy independently of whether the original images were recorded on film and scanned or recorded directly on a digital medium such as the charge coupled device (CCD). As soon as one digitizes images, one encounters a range of effects and artefacts specific to the digital world such as aliasing, quantization errors, and ambiguities in the modulation transfer function. Even a quick discussion of these effects makes it clear that what comes out of a digitizer is not related in a simple way to what goes in. After introducing the commonly used recording media, I will discuss the most important effects of digitization and then compare the limitations and advantages of the different recording media for various applications. PMID- 10816264 TI - Digitally collected cryo-electron micrographs for single particle reconstruction. AB - Several advantages and disadvantages have been cited for image collection with a slow-scan CCD camera. Here we explore its use for cryo-EM single particle reconstruction and present two practical examples. The icosahedral adenovirus (Ad) type 2 ( approximately 150 MDa) was reconstructed from 396 particle images. The Fourier shell correlation (FSC) 0.5 threshold and the Fourier shell phase residual (FSPR) 45 degrees criterion yielded 17 AA resolution for the ordered viral capsid. Visual comparison with the filtered Ad2 crystallographic hexon confirmed a resolution range of 15-17 A. The asymmetric DNA-PKcs protein (470 kDa) was reconstructed from 9,473 particle images, using a previously published reconstruction based on class-sum images as an orientational search model [Chiu et al. (1998) J. Mol. Biol. 284:1075-1081]. FSC and FSPR methods yielded 17 A resolution for the new DNA-PKcs reconstruction, indicating a small but noticeable improvement over that of the class-sum based reconstruction. Despite the lack of symmetry for DNA-PKcs and its lower image contrast compared to Ad2 (0.8% vs. 2.5%), the same resolution was obtained for both particles by averaging significantly more DNA-PKcs images. Use of the CCD camera enables the microscopist to adjust the electron beam strength interactively and thereby maximize the image contrast for beam sensitive samples. On-line Fourier transformation also allows routine monitoring of drift and astigmatism during image collection, resulting in a high percentage of micrographs suitable for image processing. In conclusion, our results show that digital image collection with the YAG-scintillator slow-scan CCD camera is a viable approach for 3D reconstruction of both symmetric and asymmetric particles. PMID- 10816265 TI - Angular reconstitution of the Staphylothermus marinus phosphoenolpyruvate synthase. AB - The processes of single particle electron crystallography and three-dimensional angular reconstitution are applied to digital cryoelectron images of a macromolecular complex, the Staphylothermus marinus phosphoenolpyruvate synthase. In particular, the application of IQAD (iterative quaternionic angular determination) is exemplified in the context of more canonical approaches. PMID- 10816266 TI - Electron detection characteristics of a slow-scan CCD camera, imaging plates and film, and electron image restoration. AB - Electron detection characteristics are summarized for the slow scan CCD (SSC) camera, imaging plates, and film. The advantage of each detector is demonstrated with the selected examples of electron diffraction and imaging. The Richardson Lucy algorithm for image restoration is described and tested for images recorded with the SSC camera. The effectiveness of image restoration is demonstrated for the recorded high-resolution lattice image, energy-loss spectrum, and convergent beam electron diffraction (CBED) pattern. PMID- 10816267 TI - Characterisation of the signal and noise transfer of CCD cameras for electron detection. AB - Methods to characterise the performance of CCD cameras for electron detection are investigated with particular emphasis on the difference between the transfer of signal and noise. Similar to the Modulation Transfer Function MTF, which describes the spatial frequency dependent attenuation of contrast in the image, we introduce a Noise Transfer Function NTF that describes the transfer of the Poisson noise that is inevitably present in any electron image. A general model for signal and noise transfer by an image converter is provided. This allows the calculation of MTF and NTF from Monte-Carlo simulations of the trajectories of electrons and photons in the scintillator and the optical coupling of the camera. Furthermore, accurate methods to measure the modulation and noise transfer functions experimentally are presented. The spatial-frequency dependent Detection Quantum Efficiency DQE, an important figure of merit of the camera which has so far not been measured experimentally, can be obtained from the measured MTF and NTF. The experimental results are in good agreement with the simulations and show that the NTF at high spatial frequencies is in some cases by a factor of four higher than the MTF. This implies that the noise method, which is frequently used to measure the MTF, but in fact measures the NTF, gives over-optimistic results. Furthermore, the spatial frequency dependent DQE is lower than previously assumed. PMID- 10816268 TI - New high-DQE imaging plate scanner using the reflected readout laser signal for noise corrections. AB - Imaging Plates (IPs) are in principle ideal electron detectors combining a large active layer area with a high sensitivity, linear dynamic range detection over 5 orders of magnitude. A moderate resolution and a decreasing detection quantum efficiency (DQE) for higher electron doses limit their use so far. The decrease of the DQE results from linear noise contributed by readout laser instabilities and inhomogeneities of the IP active layer. Here we present data on a new IP drum scanner prototype. This scanner combines twin channel amplification electronics with a new type of readout laser which allows a smaller readout focus and increased stability. The current nominal pixel size is 25 microm, and the measured modulation transfer function (MTF) indicates that further reduction of the scanning step size down to pixel sizes in the range of 12-15 microm should be possible. A unique feature of the new scanner is the simultaneous recording of the reflected readout laser light. The reflected light signal can be used for a posteriori alignment of repeated scans of one individual IP and for a correction of one part of the high spatial frequency noise contribution (reflected light correction). The posteriori alignment now allows an easy conventional gain normalization of the luminescence signal without using special markers on the IP. Both corrections lead to an increase of the DQE for high electron doses. PMID- 10816269 TI - Evaluation of scanners and CCD cameras for high-resolution TEM of protein crystals and single particles. AB - The modulation transfer function (MTF) and the geometric errors of two flatbed scanners, a slow-scan CCD (SSC) camera and film, have been measured and compared. The geometric errors of the SSC camera and film have been measured using diffraction spots from a lipid crystal. The SSC camera was shown to have the smallest geometric errors while film had the best MTF. Even though film had the best MTF, this is significantly reduced when scanning the film, so that the MTF of the film and scanner combined are comparable to the MTF of the SSC camera. PMID- 10816270 TI - Exit wave reconstructions using through focus series of HREM images. AB - The through focus exit wave reconstruction technique uses a series of high resolution electron microscopy images to reconstruct the complex electron wavefunction at the exit plane of the specimen. The feasibility of the through focus exit wave reconstruction method and its most important limitations are discussed. It is shown that-provided all aberrations of the microscope are well corrected for-a large improvement in the interpretability of the information can be obtained. PMID- 10816272 TI - Carotid stenting: rethinking the investigational-regulatory strategy PMID- 10816271 TI - Carotid stenting in patients older than 65 years with inoperable carotid artery disease: a single-center experience. AB - Carotid angioplasty and stenting to treat extracranial carotid stenosis is an alternative (as yet not widely accepted) to high-risk surgery, but its safety and efficacy are little known, especially in elderly patients. We reviewed our 3-year experience of treating 100 elderly patients (> 65 years old) considered to be inoperable (76 men, 24 women, mean age 76+/-10 years, mean follow-up 18+/-9.2 months) and present two case histories. Most (85%) were symptomatic (transient ischemic attacks in 60, stroke in 25); 80 had concomitant coronary artery disease (severe in 30 [defined by > 70% stenosis in two or more epicardial coronary arteries or the left main coronary artery]) and 25 had severe left ventricular dysfunction (ejection fraction < or =20%). The procedure was technically successful in all patients; there was one major stroke and no patient died. Postprocedure, 15% had minor complications: reversible neurological deficit (5%), pulmonary edema (3%), prolonged hypotension (3%), vascular access complications (3%), and neck hematoma (1%). Over 90% of patients were discharged home within 24 hr. PMID- 10816273 TI - Electromechanical mapping of regional left ventricular function in humans: comparison with echocardiography. AB - A catheter-based method of mapping left ventricular electromechanical regional function may be used to optimize application of local myocardial therapies by demarcating zones of ischemia or infarction. We thus performed a detailed comparison between electromechanical parameters and segmental function as assessed by echocardiography in 10 patients (3 with normal ventricles and 7 with old infarcts). Using a 16-segment model, unipolar voltage and local shortening were significantly and independently related to echo score by multivariate analysis, having a concordance with echo score of 73% for shortening and 79% for voltage. Area under ROC curves, expressing the ability to differentiate normal from abnormal segments, had values of 0.75 and 0.81 for local shortening and unipolar voltage, respectively. In conclusion, automatic assessment of regional ventricular function can be achieved independently by electrical and mechanical parameters, compared with echocardiography, permitting an integrated approach to the evaluation of ventricular function and aiding localization of catheter-based therapies. PMID- 10816275 TI - Bigger is better but longer is a limitation PMID- 10816274 TI - Importance of lesion length on new device angioplasty of native coronary arteries. NACI Investigators. New Approaches to Coronary Interventions. AB - The influence of lesion length on early and late outcomes after new device angioplasty has not been well documented. We reviewed the clinical and angiographic outcomes of 2,980 patients (3,902 lesions) undergoing new device angioplasty of native vessels enrolled in the New Approaches to Coronary Interventions (NACI) Registry. Patients were divided into three groups according to the longest lesion length (< 10, 10-20, and > 20 mm) treated. Patients with the longest lesions had more multivessel disease (56.9% vs. 49.0%, P<0.05), right coronary artery disease (52.7% vs. 32.0%, P<0.001), and total occlusions (19.1% vs. 2.5%, P<0.001) than patients with shorter lesions. Longest lesions had the smallest minimal lumen diameter (P<0.001) at baseline and at the end of the procedure. Although in-hospital events were similar, there were differences in clinical outcomes at 1 year due mainly to more target lesion revascularization in the longest lesion group (P<0.01). Multivariate analysis showed that each 1-mm increase in lesion length was associated with an increase relative risk of 1.014 (95% CI, 1.004-1.025) for target lesion revascularization at 1 year. We conclude that despite similar early clinical events, patients undergoing new device angioplasty of longer lesions have more target lesion revascularization at 1 year. PMID- 10816276 TI - Balloon angioplasty of coarctation of the aorta: a safe alternative for surgery in adults: immediate and mid-term results. AB - Patients with coarctation of the aorta can be treated either with surgery or with balloon angioplasty. So far, the last method has proved to be successful in children, but results of this treatment in (young) adults are virtually unknown. The aim of this study was to evaluate the immediate and mid-term follow-up results of balloon angioplasty of native coarctation in (mainly young) adults. Coarctation of the aorta was diagnosed by means of ultrasound or angiography, and defined as a stenosis with a pressure gradient greater than 20 mm Hg. The balloon angioplasty-procedure was carried out under complete anesthesia, and was considered to be successful, if the pressure gradient was reduced to less than 20 mm Hg. Nineteen consecutive adults (12 males, 7 females; aged 14-67 years, median 29) with native coarctation were treated from 1995-99. Mean pressure gradient decreased from 49.3+/- 20.8 to 4.8+/-8.2 mm Hg (P<0.0001). One patient showed a suboptimal result with a residual pressure gradient of 28 mm Hg. In one other patient a stent was placed on request of the referring physician. Follow-up was 100% complete and ranged from 3-47 months (mean 20.2+/- 12.9). At 1-year follow up mean systolic blood pressure was reduced from 159.4+/-19.5 to 132.5+/-17.6 mm Hg (n = 18; P<0.0001), and mean ankle-arm pressure index improved from 0.73+/ 0.09 to 0.96+/-0.05 (n = 18; P<0.0001). Anti-hypertensive medication could either be reduced or stopped in 7 patients (53.8%). With ultrasound or angiography or MRI, no patients had signs of aneurysm formation or worsening restenosis during follow-up. In adult patients with uncomplicated native coarctation of the aorta, balloon angioplasty (without stenting) would seem to be an excellent and safe alternative for surgery. In our hospital it has completely replaced surgical correction in such patients. PMID- 10816277 TI - Predictive factors for the second restenosis after coronary interventions. AB - One of the major limitations in coronary intervention is restenosis. This study was aimed to identify clinical, angiographic, and procedural factors that may be related to the second restenosis (SR). We studied 101 patients who underwent more than two follow-up coronary angiograms after two coronary interventions between January 1996 and December 1998 in Chonnam University Hospital (out of 4,092 total coronary interventions in 3,030 patients during the same period). The patients were divided into two groups according to the evidence of SR. Fifty-two patients (group A: 57+/-10 years, M:F = 44:8) who had SR and the other 49 patients (group B: 54+/-9 years, M:F = 44:5) without SR were analyzed. Clinical features, angiographic characteristics, coronary interventional procedures, and other risk factors were compared between two groups by univariate analysis and multivariate stepwise logistic regression analysis was performed for the predictive factors for SR. The clinical variables of age, sex, clinical diagnosis, and risk factors were not different between two groups. The lesion severer than B(2) by AHA/ACC classification were associated with SR (P<0.05). Recurrent angina as an indication for follow-up angiography was associated with SR (P<0.01). Predictive factors associated with SR were patient's subjective symptom and lesion severer than type B(2) according to AHA/ACC classification. PMID- 10816278 TI - Clinical and angiographic follow-up after single long GFX coronary stent implantation. AB - In order to identify predictors of late restenosis after GFX stent implantation, procedural and 6-month clinical and angiographic follow-up data of prospectively entered 141 consecutive lesions treated with a single long (24 or 30 mm) GFX stent were compared to 66 consecutive lesions requiring a single short (12 or 18 mm) stent. The initial clinical success rate of 97% and thrombosis rate of 1.4% with long stents were similar to 97% and 0% with short stents (P = NS). Their respective binary restenosis rates were 34.7% and 23.3% for long and short stents as a whole (P = NS), but being 10.0% for 12 mm, 26.0% for 18 mm, 31.3% for 24 mm, and 39.2% for 30 mm. When proximal and distal reference diameters at baseline were compared between the lesions with and without restenosis, proximal reference diameters were not statistically different (3.02+/-0.42 mm vs. 3.18+/-0.62 mm) and the restenosis group had significantly smaller distal reference diameters (2.15+/-0.48 mm vs. 2.55+/-0.53 mm, P<0.0001). The treatment of long lesions with single long-stent implantation can be accomplished with high success and low complication rates. Single long-stent implantation may be effective, if the distal reference size of the long narrowing is big enough to accept the stent. PMID- 10816279 TI - Primary transcatheter treatment of congenital pulmonary arteriovenous malformation causing cyanosis of the newborn. AB - We report on successful transcatheter therapy of congenital pulmonary arteriovenous malformation resolving persistent cyanosis and obviating the need for surgical pneumonectomy or lobectomy. PMID- 10816280 TI - Pulmonary arteriovenous malformations: transcatheter embolization options. PMID- 10816281 TI - Balloon arterioplasty of recurrent coarctation after the modified Norwood procedure in infants. AB - Coarctation of the reconstructed aorta after the modified Norwood procedure for hypoplastic left heart syndrome is a relatively common occurrence that can have significant adverse effects. From 1992-98, 65 infants with hypoplastic left heart syndrome or variants thereof underwent a modified Norwood procedure. Of the 50 survivors of Stage I palliation, 10 (20%) were subsequently diagnosed with clinically significant obstruction of the aortic arch at a median age of 5.1 months. Eight of these patients underwent coarctation balloon arterioplasty. All 8 patients underwent successful dilation of the coarctation, with a decrease in the pressure gradient from 43+/-20 to 4+/-5 mm Hg and an increase in aortic diameter at the site of obstruction from 3.5+/-0.5 to 7.0+/-1.0 mm (both P<0.001). There was also a significant increase in lower extremity systolic blood pressure. Although there was no immediate increase in cardiac output after dilation in most patients, ventricular function was improved in 3 patients within 24-48 hr of the procedure and in all but 2 at the most recent follow-up (median 2 years). Two patients developed bradycardia during wire placement that required cardiopulmonary resuscitation. Two others died suddenly during the follow-up period. All surviving patients have undergone subsequent palliative procedures; none have required repeat intervention for arch obstruction. We found a 20% incidence of coarctation in survivors of the modified Norwood procedure. Balloon arterioplasty is effective in relieving this obstruction, and should be carried out promptly upon diagnosis to minimize its deleterious effects on ventricular function. Balloon dilation, however, should be considered a high-risk procedure in these fragile infants, and special attention must be given to cardiovascular support. PMID- 10816282 TI - Pulmonary thrombosis, homocysteinemia, and reperfusion edema in an adolescent. AB - Deep vein thrombosis, pulmonary embolism, and pulmonary thrombosis in situ are rare in childhood and adolescence [1,2]. Unfortunately, these diagnoses may be unsuspected in a pediatric patient with dyspnea and chest pain. This article illustrates the diagnostic and therapeutic challenges that arose from unrecognized chronic thrombotic disease in an adolescent. PMID- 10816283 TI - Alteration of coronary flow velocity during spontaneous angina in a patient with microvascular angina. AB - Phasic coronary flow velocity in the left and right coronary artery was recorded in a patient with microvascular angina. Coronary flow velocity during anginal attack was characterized by diminished systolic forward velocity, the appearance of systolic flow reversal, increase in diastolic flow velocity and its rapid deceleration. It was also accompanied with abnormal coronary flow reserve. These findings completely recovered at follow-up examination. A subgroup of patients with microvascular angina may show unique and reversible coronary flow abnormalities during chest pain. PMID- 10816284 TI - Transcatheter treatment of a case of Lutembacher syndrome. AB - Lutembacher syndrome refers to the rare combination of congenital atrial septal defect and acquired mitral stenosis. This condition is usually treated by mitral valve operation with concomitant closure of the atrial septal defect. We describe a case of Lutembacher syndrome that was treated successfully with percutaneous transcatheter mitral commissurotomy using the Inoue balloon and closure of the atrial septal defect with the Amplatzer atrial septal defect occluder. PMID- 10816285 TI - Staphylococcal pericarditis following percutaneous transluminal coronary angioplasty. AB - Infectious complications occurring after percutaneous transluminal coronary angioplasty are uncommon. We are reporting a case of bacterial pericarditis developing 1 week after coronary angioplasty and stent implantation. Treatment with appropriate antibiotics and drainage of the infected pericardial effusion was followed by a protracted hospital course and eventual control of infection and discharge of the patient. PMID- 10816286 TI - Fatal left main coronary artery embolism from aortic valve endocarditis following cardiac catheterization. AB - Coronary artery embolization has been associated with sudden cardiac death. It is more commonly seen with aortic valve endocarditis. It manifests as acute myocardial ischemia or infarction, causing instability of the cardiac rhythm, which may be fatal. We report a patient with aortic valve endocarditis who had sudden cardiac death following coronary angiography. Autopsy revealed embolic occlusion of the left main coronary artery. PMID- 10816287 TI - Endoluminal treatment of acute aortoiliac thrombosis. AB - A 55-year-old woman presented with an ulcerative stenosis of the distal abdominal aorta. She was scheduled to undergo percutaneous aortic balloon angioplasty the next day. Upon readmission, however, she complained of lower abdominal pain. A second abdominal angiogram revealed thrombosis of the infrarenal abdominal aorta and left common iliac artery. This was treated by means of simple transcatheter thrombus aspiration and thrombolysis, followed by stenting of the aorta. PMID- 10816288 TI - Membrane-covered stents for the treatment of aortocoronary vein graft disease. AB - We report of a 78-year-old patient who underwent angioplasty for two de novo lesions in different aortocoronary venous bypass grafts. Whereas one lesion was treated by conventional stents, two adjacent lesions in a second bypass graft were covered by two newly designed stents with a polytetrafluorethylene (PTFE) membrane in between two layers of struts. The patient was first readmitted at month 5 with recurrent angina pectoris. High-grade in-stent restenosis of the lesion covered by the conventional stent was diagnosed, whereas the lesions treated by the membrane-covered stent revealed no restenosis. The patient underwent subsequent balloon angioplasty and was readmitted with unstable angina 8 months after the initial procedure. The angiogram revealed subtotal occlusion of the vein graft due to diffuse in-stent restenosis within the conventional stent. Undergoing re-PTCA, the patient received an additional membrane-covered stent placed over the proximal device. For the following 7 months, the patient stayed asymptomatic and underwent elective reangiography, which revealed a satisfying long-term result with no relevant luminal loss at the target site. This intraindividual comparison of stents in aortocoronary vein grafts suggests that PTFE membrane-covered stents might reduce restenosis as compared to conventional stents in venous bypass grafts. Furthermore, these stents potentially represent a new treatment strategy to fight in-stent restenosis in selected lesions. PMID- 10816289 TI - Promises, promises: the covered stent. PMID- 10816290 TI - Ultrasound thrombolysis for the treatment of thrombotic occlusion of degenerated saphenous vein grafts. AB - Despite improvements in catheter-based revascularization outcomes, coronary interventionalists face difficult challenges in the treatment of the thrombus laden coronary lesion. In this report, we describe the use of the Acolysis device, which utilizes high-frequency (41.9 kHz) ultrasonic energy to vibrate a small metal tip at the end of a 4.5 Fr catheter to treat two thrombotically occluded saphenous vein grafts in two patients. In both cases, the Acolysis device provided normalization of flow with angiographically evident dissolution of thrombus and excellent acute angiographic and clinical results. We conclude that in these two selected cases the Acolysis device was used safely and effectively for thrombus debulking as an adjunct to stenting in diseased saphenous vein bypass grafts. PMID- 10816291 TI - Immediate sealing of arterial puncture sites after cardiac catheterization and coronary interventions: initial U.S. feasibility trial using the Duett vascular closure device. AB - The aim of this first U.S. feasibility study was to investigate the safety and efficacy of a novel vascular sealing device, Duett, following percutaneous endovascular procedures. Immediately following a catheterization procedure, the sealing device was deployed at the femoral arterial access site in 43 patients (diagnostic 29, intervention 11, intervention + abciximab 3). Patients were followed up at 1 month with clinical assessment, ankle-brachial index measurement, and Doppler ultrasound of the treated femoral artery puncture site. Successful hemostasis was achieved with the Duett alone in 42/43 (97.7%) patients. There was one uncomplicated crossover to manual compression. The time to hemostasis was 4.0+/-1.5, 6.9+/-4.2, and 5.8+/-1.2 min for diagnostic, interventional, and abciximab patients, respectively. At 1-month follow-up, one patient (2.3%) required ultrasound-guided compression for treatment of a pseudoaneurysm. There were no other major complications. This novel vascular sealing device appears to achieve rapid and safe hemostasis successfully immediately following a wide range of percutaneous endovascular procedures. PMID- 10816292 TI - Glue for the artery and tissue track. PMID- 10816293 TI - Unsuccessful long-term outcome after treatment of a vein graft false aneurysm with a polytetrafluoethylene-coated Jostent. AB - We describe a case in which a stented ulcerated plaque in an old vein graft ruptured to a huge false aneurysm. By the use of a PTFE-coated Jostent, the false aneurysm could be percutaneously closed. However, 6 months later, a new false aneurysm, probably due to focal perforation of the covered Jostent, developed. PMID- 10816294 TI - The dark side(s) of the force. PMID- 10816295 TI - Elastic recoil of coronary stents: a comparative analysis. AB - Minimum elastic recoil (ER) has became an essential feature of new coronary stents when deployed in artheromatous lesions of various morphologies. The ER of coronary stent might be an important component of 6-month restenosis rate by minimizing the luminal loss. We evaluated the intrinsic ER of 23 coronary stents with a mechanical test bench. The amount of ER for one size of stent (3.0 mm) was quantified using a 3D optical contactless machine (Smartscope MVP, Rochester, NY). The stents were expanded on their own balloon for the precrimped stents; the uncrimped stents were expended using identical 3.0-mm balloons. Two types of measurements were done without exterior stress and with a 0.2-bar exterior stress, directly on the stent at the end of balloon expansion, immediately after balloon deflation, and then 30 min, 60 min, and 120 min after. ER ranged from 1.54%+/-0.81% (Bestent BES 15) to 16.51%+/-2.89% (Paragon stent) without stress (P<0.01) and from 2.35%+/-1.14% (Bestent BES 15) to 18.34%+/-2.41% (Cook GR2) under 0.2-bar pressure (P<0.0001). Furthermore, there was a significant reduction between the mean result of tubular stents (TS) and coil stents (CS). The results of in vitro mechanical tests may confirm strongly the interest of a minimum ER in the prevention of the 6-month restenosis. PMID- 10816296 TI - Stent longitudinal flexibility: a comparison of 13 stent designs before and after balloon expansion. AB - Longitudinal flexibility is an important property of coronary stents, facilitating delivery and allowing the expanded stent to conform to vessel contour. Subjective descriptions of flexibility abound, but there are few independent quantitative data to aid stent selection. A three-point bend test was employed to measure stiffness, the reciprocal of flexibility, for 13 stent designs in the unexpanded (bare) state, then after expansion with a 3.5-mm balloon. For eight of the designs, stiffness of the proprietary stent/balloon delivery system was also measured. In the unexpanded state, there was a wide spread of stiffness, which ranged from 0.5+/-0.2 to 91.5+/-10.0 g force/mm, depending on design. Stiffness was least for the coil (Wiktor and Crossflex) and hybrid (AVE GFX and Bard XT) designs. The MultiLink was the most flexible and the Crown the stiffest of the slotted tube designs. All stents became stiffer upon expansion. For most manufacturer-mounted stents, the delivery balloon was the main determinant of stent/balloon delivery system stiffness. Manufacturer-mounted stent profile ranged from 1.15+/-0.11 mm for the Jostent to 1.53 +/- 0.05 mm for the MultiLink system. Independent quantitative assessment of characteristics such as flexibility and profile should aid rational comparison of stent designs. PMID- 10816297 TI - Infant buttoned device. PMID- 10816298 TI - Report of "subclinical aortic perforation" with the infant double-button (Sideris) patent ductus occluder was most likely operator related. PMID- 10816299 TI - Abstracts PMID- 10816301 TI - Legal challenges to animal experimentation. PMID- 10816302 TI - A new slant on the development of orientation selectivity. PMID- 10816303 TI - AMPA receptors jump the synaptic cleft. PMID- 10816304 TI - Designer genes and anti-anxiety drugs. PMID- 10816305 TI - Sighs and gasps in a dish. PMID- 10816306 TI - Rapid and selective induction of BDNF expression in the hippocampus during contextual learning. AB - The hippocampus is required for many forms of long-term memory in both humans and animals, and formation of long-lasting memories requires the synthesis of new proteins. Furthermore, the long-term potentiation (LTP) of hippocampal synapses, a widely studied model of memory, also depends on both de novo gene transcription and protein synthesis and results in the activation of transcription from promotors containing the cAMP response element (CRE). Expression of several genes is induced during the establishment of LTP; these include the immediate-early genes (IEGs) BDNF (brain-derived neurotrophic factor), zif268 and C/EBPbeta (CCAAT-enhancer binding protein beta), all of which contain CRE sites within their promotor regions. However, these genes induced by LTP are not known to be rapidly induced following learning in a natural setting. Here we demonstrate rapid and selective induction of BDNF expression during hippocampus-dependent contextual learning. PMID- 10816307 TI - An oblique effect in human primary visual cortex. AB - Visual perception critically depends on orientation-specific signals that arise early in visual processing. Humans show greater behavioral sensitivity to gratings with horizontal or vertical (0 degrees /90 degrees; 'cardinal') orientations than to other, 'oblique' orientations. Here we used functional magnetic resonance imaging (fMRI) to measure an asymmetry in the responses of human primary visual cortex (V1) to oriented stimuli. We found that neural responses in V1 were larger for cardinal stimuli than for oblique (45 degrees /135 degrees ) stimuli. Thus the fMRI pattern in V1 closely resembled subjects' behavioral judgments; responses in V1 were greater for those orientations that yielded better perceptual performance. PMID- 10816308 TI - Cell replacement therapies for central nervous system disorders. AB - In animal models, immature neural precursors can replace lost neurons, restore function and promote brain self-repair. Clinical trials in Parkinson's disease suggest that similar approaches may also work in the diseased human brain. But how realistic is it that cell replacement can be developed into effective clinical therapy? PMID- 10816309 TI - Synaptic plasticity and dynamic modulation of the postsynaptic membrane. AB - The biochemical composition of the postsynaptic membrane and the structure of dendritic spines may be rapidly modulated by synaptic activity. Here we review these findings, discuss their implications for long-term potentiation (LTP) and long-term depression (LTD) and propose a model of sequentially occurring expression mechanisms. PMID- 10816310 TI - Synaptic activation of AMPA receptors inhibits GABA release from cerebellar interneurons. AB - A single neurotransmitter elicits diverse physiological responses through activation of multiple receptor subtypes and/or heterosynaptic interactions involving distinct synaptic targets. We found that a typical excitatory transmitter released from the climbing fiber (CF) in the cerebellar cortex not only excited Purkinje cells directly but also presynaptically inhibited GABAergic transmission from interneurons converging on the same Purkinje cells. Both homosynaptic and heterosynaptic actions of the CF transmitter (possibly glutamate) were mediated by activation of AMPA receptors. Dual AMPA receptor mediated functions of excitation and disinhibition may ensure transmission of cerebellar CF signals controlling sensorimotor coordination. PMID- 10816311 TI - Cyclic GMP-dependent feedback inhibition of AMPA receptors is independent of PKG. AB - In central neurons, the second messenger cGMP is believed to induce long-term changes in efficacy at glutamatergic synapses through activation of protein kinase G (PKG). Stimulating nitric oxide synthase, activating soluble guanylyl cyclase or elevating concentrations of intracellular cGMP depressed excitatory synaptic transmission in CA1 hippocampal neurons. Unexpectedly, intracellular cGMP depressed responses of AMPA receptors and inhibited excitatory postsynaptic currents in hippocampal neurons independently of phosphorylation. Our findings demonstrate that cGMP's modulation of excitatory transmission may involve a coupling of AMPA channel activity to levels of cGMP. PMID- 10816312 TI - Tracking presynaptic Ca2+ dynamics during neurotransmitter release with Ca2+ activated K+ channels. AB - Neurotransmitter release during action potentials is thought to require transient, localized [Ca2+]i as high as hundreds of micromolar near presynaptic release sites. Most experimental attempts to characterize the magnitude and time course of these Ca2+ domains involve optical methods that sample large volumes, require washout of endogenous buffers and often affect Ca2+ kinetics and transmitter release. Endogenous calcium-activated potassium (KCa) channels colocalize with presynaptic Ca2+ channels in Xenopus nerve-muscle cultures. We used these channels to quantify the rapid, dynamic changes in [Ca2+]i at active zones during synaptic activity. Confirming Ca2+-domain predictions, these KCa channels revealed [Ca2+]i over 100 microM during synaptic activity and much faster buildup and decay of Ca2+ domains than shown using other techniques. PMID- 10816313 TI - Excitatory peptides and osmotic pressure modulate mechanosensitive cation channels in concert. AB - Behavioral and neuroendocrine responses underlying systemic osmoregulation are synergistically controlled by osmoreceptors and neuropeptides released within the hypothalamus. Although mechanisms underlying osmoreception are understood, the cellular basis for the integration of osmotic and peptidergic signals remains unknown. Here we show that the excitatory effects of angiotensin II, cholecystokinin and neurotensin on supraoptic neurosecretory neurons are due to the stimulation of the stretch-inactivated cation channels responsible for osmoreception. This molecular convergence underlies the facilitatory effects of neuropeptides on responses to osmotic stimulation and provides a basis for the gating effects of plasma osmolality on the responsiveness of osmoregulatory neurons to peptidergic stimulation. PMID- 10816314 TI - Overexpression of Math1 induces robust production of extra hair cells in postnatal rat inner ears. AB - For mammalian cochlear hair cells, fate determination is normally completed by birth. We report here that overexpression of Math1, a mouse homolog of the Drosophila gene atonal, in postnatal rat cochlear explant cultures resulted in extra hair cells. Surprisingly, we found that the source of the ectopic hair cells was columnar epithelial cells located outside the sensory epithelium in the greater epithelial ridge, which normally give rise to inner sulcus cells. Moreover, Math1 expression also facilitated conversion of postnatal utricular supporting cells into hair cells. Thus Math1 was sufficient for the production of hair cells in the ear, and immature postnatal mammalian inner ears retained the competence to generate new hair cells. PMID- 10816315 TI - Sedative but not anxiolytic properties of benzodiazepines are mediated by the GABA(A) receptor alpha1 subtype. AB - Inhibitory neurotransmission in the brain is largely mediated by GABA(A) receptors. Potentiation of GABA receptor activation through an allosteric benzodiazepine (BZ) site produces the sedative, anxiolytic, muscle relaxant, anticonvulsant and cognition-impairing effects of clinically used BZs such as diazepam. We created genetically modified mice (alpha1 H101R) with a diazepam insensitive alpha1 subtype and a selective BZ site ligand, L-838,417, to explore GABA(A) receptor subtypes mediating specific physiological effects. These two complimentary approaches revealed that the alpha1 subtype mediated the sedative, but not the anxiolytic effects of benzodiazepines. This finding suggests ways to improve anxiolytics and to develop drugs for other neurological disorders based on their specificity for GABA(A) receptor subtypes in distinct neuronal circuits. PMID- 10816316 TI - Motor coordination without action potentials in the mammalian spinal cord. AB - Coordination of neuronal activity to produce movement is generally thought to depend on spike activity in premotor interneuronal networks. Here we show that even without such activity, the neonatal rat spinal cord could produce a stable motor rhythm mediated by the synchronization of motor neuron oscillations across gap junctions. These rhythms, however, were not coordinated between motor pools in different parts of the spinal cord. We further showed that neural coordination through gap junctions contributed to the fundamental organization and function of spinal motor systems. These results suggest that neural coordination across gap junctions is important in motor-pattern generation in the mammalian spinal cord. PMID- 10816317 TI - Reconfiguration of the neural network controlling multiple breathing patterns: eupnea, sighs and gasps [see comment]. AB - Are different forms of breathing derived from one or multiple neural networks? We demonstrate that brainstem slices containing the pre-Botzinger complex generated two rhythms when normally oxygenated, with striking similarities to eupneic ('normal') respiration and sighs. Sighs were triggered by eupneic bursts under control conditions, but not in the presence of strychnine (1 microM). Although all neurons received synaptic inputs during both activities, the calcium channel blocker cadmium (4 microM) selectively abolished sighs. In anoxia, sighs ceased, and eupneic activity was reconfigured into gasping, which like eupnea was insensitive to 4 microM cadmium. This reconfiguration was accompanied by suppression of synaptic inhibition. We conclude that a single medullary network underlies multiple breathing patterns. PMID- 10816318 TI - Diverse receptive fields in the lateral geniculate nucleus during thalamocortical development. AB - Most models of thalamocortical development in the visual system assume a homogeneous population of thalamic inputs to the cortex, each with concentric on- or off-center receptive fields. To test this, we made high-resolution spatial maps of receptive fields in the developing ferret lateral geniculate nucleus (LGN). Developing receptive fields (RFs), had a variety of shapes: some concentric, others elongated (like adult cortical receptive fields) and some with 'hot spots' of sensitivity. These receptive fields seemed to arise from convergence of multiple retinal afferents onto LGN neurons. We present a Hebbian model whereby imprecise retinogeniculate connections help refine geniculocortical connections, sharpening both thalamocortical topography and perhaps orientation selectivity. PMID- 10816319 TI - Stimulus dependence of two-state fluctuations of membrane potential in cat visual cortex. AB - Membrane potentials of cortical neurons fluctuate between a hyperpolarized ('down') state and a depolarized ('up') state which may be separated by up to 30 mV, reflecting rapid but infrequent transitions between two patterns of synaptic input. Here we show that such fluctuations may contribute to representation of visual stimuli by cortical cells. In complex cells of anesthetized cats, where such fluctuations are most prominent, prolonged visual stimulation increased the probability of the up state. This probability increase was related to stimulus strength: its dependence on stimulus orientation and contrast matched each cell's averaged membrane potential. Thus large fluctuations in membrane potential are not simply noise on which visual responses are superimposed, but may provide a substrate for encoding sensory information. PMID- 10816320 TI - The locus of attentional effects in texture segmentation. AB - Cuing covert spatial attention can increase spatial resolution. Here we pinpointed the specific locus of this effect using texture segmentation. At the level of visual cortex, texture segmentation theoretically involves passage of visual input through two layers of spatial linear filters separated by a pointwise nonlinearity. By manipulating the textures to differentially stimulate first- or second-order filters of various scales, we showed that the attentional effect consistently varied with the latter. These psychophysical results further support the hypothesis that attention increases resolution at the attended location and are consistent with an effect of attention at stages as early as the primary visual cortex. PMID- 10816321 TI - Role of INTERLEUKIN-6 in the pathogenesis of multiple myeloma. AB - Multiple myeloma (MM) is a currently incurable disease caused by the proliferation of malignant plasma cells. Although the pathogenesis of the disease still remains unclear, recent research in the biology of MM has produced new insights into the factors that control the growth and survival of myeloma cells. Among the growth factors, interleukin-6 (IL-6) has an essential role. Evidence suggests that IL-6 is not only a growth factor, but also a survival factor in MM, inhibiting apoptosis in myeloma cells. IL-6 interacts with several factors which are involved in the pathogenesis of MM, such as adhesion molecules, tumour suppressor genes and oncogenes. Considering the essential role of IL-6, it could serve as a target for new therapeutic interventions. Neutralizing the effect of IL-6 may result in a regression of tumour progression. PMID- 10816322 TI - Ganglioside-mediated metabolic synchronization of protein synthesis activity in cultured hepatocytes. AB - An ultradian oscillation of protein synthesis was detected by synchronization of metabolic activity in rat hepatocyte cultures. This oscillation occurs in dense cultures in fresh medium, but not in sparse ones. Metabolic synchronization of sparse cultures, however, was initiated by conditioned medium or addition of 0.3 0.5 microm of a mixture of bovine brain gangliosides to fresh culture medium along with either 0.06-0.2 microm GM1 or 0.1-0.2 microm GDIa. GTIb and GDIb did not produce oscillations, nor did human liver ganglioside GM3. High expression of GM1 ganglioside determinants in hepatocytes maintained in the conditioned medium purified polyclonal antibodies to GM1 was coupled with protein synthetic oscillatory activity, i.e. metabolic synchronization. Incubation of dense cultures with GM1-antibodies for 24 h decreased the amplitude of these oscillations. In sparse cultures maintained in fresh medium where protein synthesis showed no oscillatory pattern, GM1 expression was low. PMID- 10816323 TI - Glucocorticoids enhance serum deprivation- but not calcium-induced cytotoxicity in rat C6 glioma cells. AB - While glucocorticoids have been shown to exacerbate calcium-induced neuronal damage, little is known about the effects of these hormones on calcium-induced damage to glial cells. Here we examine the effect of synthetic glucocorticoid dexamethasone on calcium ionophore A23187 and serum deprivation-induced damage to rat C6 glioma cells. Treatment of the glioma cells with A23187 reduced cell viability, similar in extent to that observed with serum deprivation. Both A23187 and serum deprivation caused cell damage without degradation of the genomic DNA into nucleosomic fragments. In addition, the reduction in cell viability caused by A23187 was not significantly altered by DEX at concentrations enhancing serum deprivation-induced cell death. These results suggest that the cytotoxic effect of A23187 on glial cells may be mediated through a mechanism different from that underlying serum deprivation-induced cell death, and that, in contrast to calcium induced neuronal damage, calcium-induced damage to glial cells is likely to be insensitive to glucocorticoids. PMID- 10816324 TI - Kinetic characterization of calcium uptake by the rat liver Golgi apparatus. AB - We carried out a kinetic characterization of the Ca(2+)active transport in the rat liver Golgi Apparatus (GA) membrane. Calcium accumulation by vesicles of a GA enriched fraction was found to be a function of both Ca(2+)and ATP-Mg concentrations, it was inhibited by 2 microm thapsigargin but not stimulated by 3 microm calmodulin. The kinetic parameter values obtained for the GA Ca(2+)pump were: J(max)of 3.96 nmol/mg min, K(m)for Ca(2+)of 0.150 microm and two K(m)'s for ATP of 1.14 microm and 519 microm. These results were almost identical to those obtained for the endoplasmic reticulum (ER) fraction, indicating that the GA Ca(2+)pump is a sarco/endoplasmic reticulum (SERCA) P-type, analogous-if not identical-to that present in the ER. PMID- 10816325 TI - Proteolytic activity in intact sheets of polarized epithelial cells as determined by a cell-permeable fluorogenic substrate. AB - The purpose of the present investigation was to develop a system for continuous evaluation of extralysosomal proteolytic activity and its regulation in polarized epithelial cells. Filter inserts containing a tight monolayer of primary cultured pig thyrocytes were placed in a thermostated aluminium block. The cell-permeable, fluorogenic calpain and proteasome substrate succinyl-Leu-Leu-Val-Tyr-7-amino-4 methylcoumarin was added to the apical buffer and fluorescence changes were continuously measured via the fibre optics of a luminometer held at a fixed distance from the cell layer. Basal proteolytic activity was reduced by 60-70% by the proteasome inhibitor lactacystin. Proteolysis was increased within a few minutes after application of Ca(2+)-mobilizing agents (ionomycin, 4-bromo-A23187, thapsigargin and maitotoxin). Forskolin and staurosporine also enhanced the proteolytic activity. We conclude that Ca(2+)mobilization, and possibly also changes of protein kinase activity, rapidly increase non-lysosomal proteolysis in the intact thyroid epithelium. PMID- 10816326 TI - GALECTIN-3 expression in differentiating human myeloid cells. AB - Galectin-3 is an endogenous mammalian carbohydrate-binding protein with affinity for ABH group carbohydrate epitopes and polylactosamine glycans present on cell surface and extracellular matrix glycoproteins. It has been shown to play a role in cell differentiation, morphogenesis, adhesion and cell proliferation regulation. Progenitor cell proliferation in bone marrow depends on stem cell factors including those modulating their adhesion to the bone marrow stroma. The present study shows that the 32 kD galectin-3 is developmentally expressed in human myeloid cells and is strongly upregulated on the cell surface of late mature myeloid cells. Despite the fact that the expression of the galectin-3 is very low in CD34+ early myeloid cell, a 70 kD protein is found by Western blotting using antibodies specific to galectin-3, exclusively in those cells. Finally, exogenous human recombinant galectin-3 binds strongly to CD34+ early myeloid cells and emphasizes granulocyte-colony stimulating factor (G-CSF) driven proliferation in vitro. PMID- 10816328 TI - Experimental models for the study of cardiovascular function and disease. AB - In the study of cardiovascular biology, both under conditions of health and disease, the investigator enjoys the availability of a vast range of experimental models ranging from man to a single molecule and beyond. There is also a vast spectrum of measurable indices of function and injury. This is particularly so in the case of myocardial ischemia, a disease which still contributes to the majority of deaths in the Western Hemisphere. Each experimental model, each species and each end-point has its own inherent advantages and disadvantages and appreciating these will help the investigator select the most appropriate study system for the particular question under investigation. This article endeavours to identify some of these strengths and weaknesses and reveals the frequently encountered paradox that the greater the amount and reproducibility of data the further removed is the model from clinical reality. Fortunately, however, an appreciation of this 'weakness' can often be exploited for the advancement of knowledge. PMID- 10816327 TI - Apoptotic and non-apoptotic modes of programmed cell death in MCF-7 human breast carcinoma cells. AB - Apoptosis is a specific mode of programmed cell death (PCD), recognized by characteristic morphological and molecular changes. Here we present evidence for a non-apoptotic type of PCD in human MCF-7 breast carcinoma cells. We used TNF alpha and tyrphostin AG213 to induce apoptotic and non-apoptotic cell death respectively in vitro. Microscopic and immunohistochemical studies, together with DNA analysis and flow cytometric analysis of p53 and bcl-2 oncogene expression, revealed some novel characteristics of non-apoptotic cell death. We show here for the first time some of the biochemical features of an experimentally induced non apoptotic PCD and emphasize the distinct biochemical events leading to apoptotic and non-apoptotic PCD. PMID- 10816329 TI - Animal models of chronic heart failure. AB - Chronic heart failure is associated with multiple pathophysiological alterations and adaptations, such as marked anatomic and biochemical changes of the myocardium, left ventricular dysfunction and dilatation, increased systemic vascular resistance, and activation of neurohumoral and cytokine systems. The use of animal models has provided a new insight into the complex pathogenesis of this syndrome and supplemented clinical experience. However, all of the animal models used have advantages and limitations, and the transfer from experimental to human heart failure needs critical evaluation. The current review will focus upon new aspects of rat and rabbit models of heart failure. PMID- 10816330 TI - The isolated blood and perfusion fluid perfused heart. AB - The isolated heart is deservedly one of the most popular experimental models in cardiovascular research, both in terms of cost and the quality and quantity of data it provides. However, it is a deceptively simple model, capable of throwing many problems in the path of the inexperienced or unwary perfuser. The following article discusses the advantages and disadvantages of the various types of isolated heart perfusion (Langendorff and working; blood and buffer perfused). We attempt to give an insight into the many factors which must be taken into consideration when first establishing these preparations, we identify the range of indices that can be measured and the potential pitfalls which, with a little care, can be readily avoided. PMID- 10816331 TI - Effects of chronic ethanol consumption on alpha-adrenergic-induced contractions and endothelium-dependent relaxations in rat thoracic aorta. AB - The effects of chronic oral administration of ethanol (7.2% daily during 24 weeks) on the contractions induced by phenylephrine (Phe) and the endothelium dependent relaxation responses to acetylcholine (ACh) were studied in rat thoracic aorta. Ethanol pretreatment significantly attenuated the contractile responses to Phe, resulting in parallel shift of the concentration-response curve to the right. EC(50)values of Phe were 64.6+/-11.2 and 95.5+/-8.5 nmol l(-1)in control and ethanol-fed rats, respectively. On the other hand, either calcium induced contractions or relaxation responses to ACh and sodium nitroprusside were similar in the vessels of the control and ethanol-treated rats. These results suggest that chronic ethanol ingestion significantly attenuates the alpha(1) adrenergic-induced contractions but does not affect the relaxation responses mediated by nitric oxide in rat aortic rings. PMID- 10816332 TI - Possible mechanism of cardioprotective effect of ischaemic preconditioning in isolated rat heart. AB - The present study is designed to investigate the mechanism of the cardioprotective effect of ischaemic preconditioning. Isolated perfused rat heart was subjected to global ischaemia for 30 min followed by reperfusion for 120 min. Coronary effluent was analysed for LDH and CK release to assess the degree of cardiac injury. Myocardial infarct size was estimated macroscopically using TTC staining. Four episodes of ischaemic preconditioning markedly reduced LDH and CK release in the coronary effluent and decreased myocardial infarct size. Administration of prazosin (alpha(1)adrenoceptor antagonist) before global ischaemia reduced the extent of ischaemia-reperfusion induced myocardial injury. The cardioprotective effect of ischaemic preconditioning was abolished by prazosin and colchicine (microtubule disaggregator). On the basis of these results, it may be concluded that the cardioprotective effects of ischaemic preconditioning may be mediated through stimulation of alpha(1)adrenoceptors and translocation of PKC. PMID- 10816333 TI - Short treatments of normotensive and hypertensive rats by angiotensin II and nitric oxide inhibitor induce an increase of noradrenaline sensitivity in isolated vena portae preparations. AB - The aim of the study was to examine the influence of a short-term treatment of conscious Wistar Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) by angiotensin II (ANG II) and by ANG II in combination with either l -NAME, HOE 140 or minoxidil on the mean arterial blood pressure (MABP) and the noradrenaline sensitivity in isolated portal vein preparations. MABP was significantly increased by ANG II treatment and ANG II plus l -NAME. However, it was slightly affected by ANG II in association with HOE 140, and significantly lowered by ANG II plus minoxidil. In control animals noradrenaline increased the frequency and the tone of contractile force. While ANG II enhanced the contractile response to noradrenaline, neither in combination with l -NAME, HOE 140 nor minoxidil prevented such an increase in the response to noradrenaline. In the presence of ergotamine, the contractile response to noradrenaline was completely blocked not only in control animals, but also in animals treated with ANG II alone or in combination with minoxidil. However, ergotamine (3 microm) failed to block completely the contractile response to noradrenaline in vessels from animals treated by ANG II in combination with l -NAME or HOE 140. These data suggest that ANG II causes an increase of noradrenaline sensitivity in the isolated portal vein of rat. l -NAME and HOE 140 unmask a contractile response to noradrenaline in the presence of ergotamine which seems to be mediated not only by alpha-adrenoceptors, but may be compensated by an endothelial relaxation. PMID- 10816334 TI - Acetazolamide and enalapril combination offers complete protection from nitric oxide-deficient stroke in stroke-prone spontaneously hypertensive rats. AB - Chronic oral administration of l -NAME precipitates stroke in stroke-prone spontaneously hypertensive rats (SHRSP). The present study investigated whether acetazolamide (an acidotic agent) given alone or in combination with an angiotensin blocker (enalapril maleate) offers any protection from NO-deficient stroke in SHRSP. We also examined whether protection from NO-deficient stroke involves activation of K(+)channels. Five-week-old SHRSP drank saline (group I), l -NAME (group II), l -NAME+enalapril (group III), l -NAME+acetazolamide (group IV), and l -NAME+enalapril+acetazolamide (group V). Within a few hours following onset of stroke, rats were attached to a blood pressure recorder. In subsequent experiments, to investigate the involvement of K(+)channels, glibenclamide and BaCl(2)(K(+)channel blockers) were included in the drinking solutions that were given to the SHRSP groups receiving l -NAME, acetazolamide and enalapril. Group I of SHRSP did not develop stroke. Group II, III and IV developed stroke in 12+/-2, 29+/-2 and 20+/-2 days, respectively. SHRSP from group V did not develop stroke. However, they died in 70+/-2 days. The glibenclamide and BaCl(2)administration failed to prevent this protection from stroke. In conclusion, concurrent administration of acetazolamide and enalapril prevents onset of NO-deficient stroke in SHRSP. These stroke-protective effects are independent of reductions in mean or systolic blood pressures and do not involve an activation of K(+)channels. PMID- 10816335 TI - Comparative anti-nociceptive, anti-inflammatory and toxicity profile of nimesulide vs nimesulide and piperine combination. AB - Piperine is an inhibitor of various hepatic and other enzymes involved in the biotransformation of drugs. Preliminary pharmacokinetic studies conducted by us suggested the increased bioavailability of nimesulide co-administered with piperine. The present study was, thus, conducted to evaluate the antinociceptive, anti-inflammatory and toxicity profile of a new nimesulide-piperine combination administered orally as compared with nimesulide alone. Antinociceptive efficacy was tested using an acetic acid writhing test and tail flick latency test (TFL). The ED50 value of a nimesulide-piperine combination in writhing test was calculated to be significantly lower (1.5 mg kg(-1)) as compared to (11.2 mg kg( 1)) of nimesulide alone. The antinociceptive effect was lesser in the tail flick latency test as compared to what was observed in the writhing test indicating the peripheral action of the Non-Steriodal Anti-Inflammatory Drug (NSAID). In carrageenan-induced inflammatory tests, the nimesulide-piperine combination was found to be dose-to-dose superior than nimesulide alone. Acute toxicity studies on mice revealed a reduction in lethal dose (LD50) of the combination (980 mg kg( 1)) as compared to nimesulide (1500 mg kg(-1)) alone. Results from the present study suggest a better therapeutic index for the nimesulide-piperine combination indicating that this combination would further reduce the frequency of adverse effects associated with nimesulide alone. PMID- 10816336 TI - Withania somnifera root extract in the regulation of lead-induced oxidative damage in male mouse. AB - The importance of Withania somnifera root extract in the regulation of lead toxicity with special reference to lipid peroxidative process has been investigated in liver and kidney tissues. While lead treatment (0.5 mg kg(-1)body wt. day(-1)for 20 days) enhanced hepatic and renal lipid peroxidation (LPO), administration of plant extract in the doses of 0.7 g kg(-1)and 1.4 g kg(-1)body wt. day(-1)along with equivalent doses of lead acetate for 20 days significantly decreased LPO and increased the activities of antioxidant enzymes, viz., superoxide dismutase (SOD) and catalase (CAT), thus retaining normal peroxidative status of the tissues. We suggest that the ameliorating role of root extract of W. somnifera in the lead intoxicated mice could be the result of its antiperoxidative action. PMID- 10816337 TI - Changes in histamine content following pharmacologically-induced mast cell degranulation in the rat conjunctiva. AB - Compound 48/80 was applied into one eye of male Wistar rats and a drop of vehicle into the contralateral eye. Another group of rats received sodium cromoglycate in both eyes every 6 h for a period of 48 h. One eye was challenged with compound 48/80 30 min after the end of treatment with sodium cromoglycate. The eyes were monitored clinically and the histamine content of the conjunctiva was determined fluorometrically. The basal histamine levels in rat conjunctival homogenates were quantified. Pharmacologically-induced mast cell degranulation by a single application of 0.1 g ml(-1)of compound 48/80 resulted in significant decreases of conjunctival histamine levels 1, 12 and 24 h after challenge. Sodium cromoglycate prevented the effect of compound 48/80 when administered into the eye prior to the challenge with the non-immunogenic histamine releaser. Upon termination of the application, the membrane stabilizer was unable to reverse the reduced histamine levels in the conjunctival homogenates. PMID- 10816338 TI - Excitatory and inhibitory effects of second-generation cholinesterase inhibitors on rat gastrointestinal transit. AB - We investigated the influence of increasing oral doses of second generation acetylcholinesterase inhibitors (AChEI) such as tacrine (0.25, 0.5, 1, 2, 3, 4, 10, and 20 mg kg(-1)), eptastigmine (0.5, 4, 8, 12, 20 and 40 mg kg(-1)) and E2020 (0.18, 0.25, 0.5, 1, 2, 3, 4 and 10 mg kg(-1)) on the distance travelled by a charcoal meal administered 30 min after each compound, in comparison with physostigmine (0.5, 1, 2, 4, 8 and 12 mg kg(-1)). An inverted U regression was observed with a significant parabola between the centimetres travelled and the log of the doses for all AChEI. The maximal stimulating doses (mg kg(-1)) were 2 for physostigmine, 4 for eptastigmine, 3 for tacrine and E2020, while the inhibitory doses were 12 for physostigmine, 40 for eptastigmine, 20 for tacrine and 10 for E2020. The stimulating and inhibiting effects on gastrointestinal propulsion were significantly reversed by 0.25 mg kg(-1)of scopolamine hydrobromide. A dose of scopolamine hydrobromide (0.06 mg kg(-1)) or methylbromide (0.25 mg kg(-1)), pirenzepine dihydrochloride (0.25 mg kg(-1)) and mecamylamine hydrochloride (0.5 mg kg(-1)), which per se did not affect gastrointestinal propulsion, antagonized both the stimulating and inhibitory effect of eptastigmine. Thus, the biphasic effect is peripherally mediated through both muscarinic (at least M(1)) and nicotinic receptors. PMID- 10816340 TI - Volume contents and index PMID- 10816339 TI - Comprehensibility of the package leaflets of all medicinal products for human use: a questionnaire survey about the use of symbols and pictograms. AB - Directive 92/27EEC establishes that the package leaflet is a document, which must be included in the package of medicinal products for human use in EU countries. This informative leaflet is directed at the users and it must give full and comprehensible information. The Law suggests the use of symbols but it does not give advice about the subjects to be represented. In order to evaluate the attitude of patients towards package leaflets provided with symbols, we planned a survey interviewing 1004 patients in pharmacies. The data suggest that Italian patients usually read the package leaflet but they neither understand it easily nor find the needed information readily. Most respondents (74.3%) considered the use of symbols helpful in finding the needed information. We proposed five symbols for each heading (therapeutic indications, side effects, paediatric use, contraindications, use in pregnancy) and asked to select which symbol could be used. In the case of 'side effects', 'paediatric use', 'use in pregnancy' and 'dose', most of the respondents chose the same symbol. In the case of 'therapeutic indications' and 'contraindications' there was no uniformity in the answers. The choice depends greatly on education, age and employment of respondents. PMID- 10816341 TI - Factors affecting improvement on the American Board of Surgery In-Training Exam (ABSITE). AB - BACKGROUND: American Board of Surgery In-Training Exam (ABSITE) performance can be used to assess resident knowledge and to evaluate surgical curriculum. To determine factors that lead to improved resident ABSITE performance, a prospective study was performed. MATERIALS AND METHODS: Thirty-four surgical residents in Program Years 2-5 completed pre- and a post-ABSITE questionnaires about their anxiety, self-efficacy, physical preparation, and academic preparation for the ABSITE. Department records were used to determine resident probationary status and conference attendance. A preliminary analysis of ABSITE scores indicated a significant improvement between 1998 and 1999 percentile scores (paired t = -2. 25, P = 0.03; m = 11.9, SD = 30.5, median = 7). An improvement in percentile rank score was calculated and used as the dependent variable in a stepwise regression analysis. The following served as independent variables: previous exam performance, anxiety, probationary status, amount of sleep before exam, confidence to score in the 25th and the 50th percentiles, and attendance at the three conferences rated most valuable by the residents. RESULTS: Results of the regression analysis demonstrate that all factors account for 62.3% of the variance in improvement scores. A stepwise analysis indicated that the combination of attendance (40.2%) and previous performance (18.3%) was significant in explaining 58.5% of the variance in improvement scores. Furthermore, Pearson's correlations indicated that probationary status (+.58, P = 0.001), anxiety (+0.53, P = 0.001), amount of study (+0.61, P = 0.001), past ABSITE performance (-0.60, P = 0.001), and conference attendance (+0. 56, P = 0.001) were correlated with ABSITE improvement. CONCLUSIONS: This study demonstrates that resident individual effort, past ABSITE performance, and academic conference attendance have led to resident ABSITE improvement. PMID- 10816342 TI - Improving the Surgeon's participation in research: is It a problem of training or priority? AB - BACKGROUND: Although numerous important contributions have originated from basic science research performed by surgeons, it seems that such dedicated work is becoming increasingly difficult to accomplish. What are the reasons for this change and what improvements can be made? This study aims to characterize the basic research training and careers of senior academic surgeons to assess and devise strategies for sustaining productive and quality surgical research. METHODS: A 25-item survey was sent to 850 senior-level members of academic societies, including the Association of Academic Surgeons, Society of University Surgeons, and American Surgical Association. It addressed each surgeon's clinical and research training and career, as well as opinions concerning surgical research. RESULTS: Three hundred seventy-seven (44%) surveys were received. Mean age was 64 years, and 73% were full professors. Seventy-two percent of respondents performed basic science research during training, and for 71% of this group, research was a significant reason for choosing a clinical specialty. Ninety-one percent performed research in the same specialty area during and after training. Of those who performed research during training, a full 99% continued to perform research on completion of training. However, 38% stopped performing basic research by age 39. Seventeen and twenty-three percent stopped basic research between 40 and 49 and between 50 and 59 years of age, respectively. The most common factors causing them to stop were increased clinical load (40%) and increased administrative duties (38%). For respondents who had stopped research prior to age 40, 73% cited increased clinical load as the primary reason. Eighty five percent felt a dedicated research period should be included in surgery training. CONCLUSIONS: Most respondents had participated in basic research during training, and continued similar research after training. However, an overwhelming clinical practice at the junior faculty level seemed to hinder research. We conclude: (1) the majority consensus is that research training is integral to the development of academic surgeons; (2) such research training opportunities appear adequate; however, (3) faculty performing research, particularly at the junior level, need to be better protected from other academic duties, such as clinical practice and administration. The challenge to the leadership of academic surgery will be to enhance such research productivity in the context of increasing academic demands. PMID- 10816343 TI - Peptide YY and vitamin E inhibit hormone-sensitive and -insensitive breast cancer cells. AB - We have shown that peptide YY, an endogenous gut hormone, and vitamin E succinate (VES) inhibit pancreatic cancer cell growth in vitro. We hypothesized that PYY and VES would inhibit breast cancer cell viability regardless of the hormone receptor status. Human breast ZR-75 ductal carcinoma (estrogen receptor negative) and MCF-7 adenocarcinoma (estrogen receptor positive) cells were cultured and exposed to VES (10 pg/ml), PYY (500 pmol), or both agents together. MTT assay was performed at 24, 48, and 72 h to evaluate cell viability. At every time interval, PYY and VES significantly inhibited cell growth compared to control. The effects of PYY were similar in magnitude to those of VES. Combining the agents resulted in a significant additive inhibition of growth with the greatest effect seen at 72 h. We have shown that PYY and vitamin E inhibit in vitro growth of breast cancer cells with variable hormone receptor status. When used in combination, the agents have a significant increase in effect. Further studies are ongoing to define the mechanism of action of these agents and to translate the experiments to an in vivo model. PMID- 10816344 TI - Amino acid uptake and regulation in multicellular hepatoma spheroids. AB - BACKGROUND: Cancer cells maintained in monolayer tissue culture are frequently used to study tumor biology and nutrient uptake, but there is a concern that this system may not fully reflect clinical tumor physiology. Because cells grown in a 3-D configuration more closely resemble an in vivo environment, a model was developed and characterized for the growth of SK-Hep human hepatoma cells in suspension as multicellular tumor spheroids (MTS). The measurement of nutrient uptake in such a system has never been established. MATERIALS AND METHODS: SK-Hep cultures were initiated as single cell suspensions and grown as MTS in siliconized spinner flasks. The transport of several individual amino acids (arginine, glutamate, leucine, alpha-(N-methylamino)isobutyric acid (MeAIB), and glutamine (GLN)) was measured in SK-Hep single cell suspensions and MTS (0. 50 0.60 mm diameter) by a radiotracer/rapid filtration technique, as was the regulation of glutamine uptake by phorbol esters. l-[(3)H]GLN uptake was also measured in larger spheroids (0.85-1.5 mm diameter). MTS cellularity was evaluated by histological examination, and single cell integrity after the transport assay was confirmed by scanning electron microscopy (SEM). RESULTS: SK Hep MTS displayed gradients of cellular morphology and staining, with central necrosis visible at diameters >0.8 mm. Single cell suspensions endured the rapid filtration technique based on functional Na(+)-dependent uptake rates and SEM analysis. Of all amino acids tested, only GLN transport rates were visibly affected by growth format. In small MTS, Na(+)-dependent GLN uptake was diminished by 40%, but was 40-53% higher in MTS >1 mm displaying central necrosis, when compared to single cell suspensions. Likewise, slight parallel changes in glutamine transporter ATB(0) mRNA levels were observed in Northern blot analysis. Finally, phorbol ester-dependent GLN transport down-regulation (by 40-50%), previously established in SK-Hep monolayers, remained operative in all cell formats tested. CONCLUSIONS: The data suggest that the tumor microenvironment differentially impacts the uptake of specific nutrients despite the conservation of key regulatory pathways. This MTS technique may prove useful for further studies on the role of nutrient transport in nascent tumor growth. PMID- 10816345 TI - The angiogenesis inhibitor, endostatin, does not affect murine cutaneous wound healing. AB - BACKGROUND: Endostatin is a potent angiogenesis inhibitor, which is currently being used in Phase I trials as an antitumor agent. The purpose of this study was to determine whether endostatin has an effect on wound healing in a murine model. MATERIALS AND METHODS: The function of endostatin was confirmed using a human microvascular endothelial cell (HMVEC) proliferation assay in which cells are treated for 4 days with growth media plus or minus endostatin. Full-thickness incisions were made on the dorsum of athymic nude mice and closed primarily with skin staples. PVA sponges were implanted in some wounds to determine vascular ingrowth. Subsequently, mice were treated with recombinant human endostatin at 20 mg/kg/day or 50 mg/kg/dose BID versus control for a total of 14 days. On Days 2, 4, 8, 12, and 16, three mice per group had serum samples drawn and were sacrificed. Perpendicular breaking strength (N) was determined using an Instron 5540 tensometer. Wound strength was determined by dividing breaking strength by wound area (N/cm(2)). Vascular density in sponges was determined using CD31 immunohistochemistry. Serum endostatin concentrations were determined using a commercially available ELISA kit. RESULTS: Endostatin caused a significant reduction of endothelial cell proliferation after 4 days compared to media alone (72%, P = 0.031). At all time points tested, there was no statistical difference in the wound-breaking strength between endostatin and control-treated mice at either the low or high dose. Serum endostatin levels were consistently 10-fold higher in endostatin-treated mice than in controls. No differences in vascular density were seen in endostatin versus control-treated mice as determined by CD31 immunohistochemistry of PVA sponges. CONCLUSION: Therapy with human endostatin does not induce a significant decrease in breaking strength of cutaneous wounds in mice. PMID- 10816346 TI - Association of artery wall hypoxia and cellular proliferation at a vascular anastomosis. AB - BACKGROUND: We hypothesize that arterial wall hypoxia incites the pathologic formation of intimal hyperplasia at an artery anastomosis. We have determined from previous studies performed in our laboratory, the oxygen tension profiles of the artery wall at various times after vascular anastomosis. The purpose of this study is to determine the rate of cellular proliferation at an artery anastomosis when the artery wall is most hypoxic. MATERIALS AND METHODS: Expanded polytetrafluoroethylene (ePTFE) grafts were placed end to end in the infrarenal aorta of 27 New Zealand white rabbits. The anastomotic aortic wall oxygen (O(2)) tensions were measured with an O(2) microelectrode in rabbits 0, 7, 14, 28, and 42 days after surgery. O(2) tensions were also measured in 4 control rabbits for comparison. 5-Bromo-2'-deoxyuridine (BrDU) was injected intraperitoneally 24 h prior to rabbit sacrifice. After O(2) tension measurements, the rabbits were sacrificed and the aortic grafts were harvested. Bioquant morphometrics was used to measure cells with BrDU counterstaining and intimal thickness in 17 rabbits: in control (n = 4), Day 0 (n = 4), 7 (n = 5), and 42 (n = 4). Student's t test was used to compare O(2) tensions, cellular proliferation, and intimal hyperplasia between days. RESULTS: The pO(2) levels at the outer layers of the aorta, 1 mm distal to the distal aortic graft anastomosis, were 61.0 +/- 2 (+/ SE) mm Hg for controls, 19.8 +/- 1 mm Hg for Day 7 (P < 0.0001), 19.0 +/- 1 mm Hg for Day 14, 39.2 +/- 1 mm Hg for Day 28, and 58.5 +/- 1 mm Hg for Day 42 aortic grafts. BrDU-staining ratios in the intima were significantly higher in the Day 7 aortic grafts, 28.6 +/- 3%, versus BrDU-staining ratio, 1.4 +/- 1%, in Day 42 aortic grafts (P < 0.0002). CONCLUSIONS: Cellular proliferation is highest at Day 7 when the artery wall is most hypoxic and returns to baseline as O(2) tensions normalize. PMID- 10816347 TI - Epidermal growth factor alters the bax:bcl-w ratio following massive small bowel resection. AB - BACKGROUND: Following massive small bowel resection (SBR), the expression of bax and bcl-w is associated with increased enterocyte apoptosis. Epidermal growth factor (EGF) has been shown to enhance enterocyte proliferation and retard apoptosis in the adapting bowel. This study examined the effect of EGF on the expression of these bcl-2 family members during adaptation. MATERIALS AND METHODS: Mice (C57Bl/6; n = 38) underwent a 50% SBR or sham operation and then were randomized to receive twice-daily orogastric saline or EGF (50 microg/kg/day). After 3 days, the remnant ileum was removed, apoptotic index (No. apoptotic bodies/crypt) calculated, and expression of mRNA and protein for bax and bcl-w quantified. RESULTS: EGF prevented the expected increase in the apoptotic index after SBR and altered the ratio of bax to bcl-w in favor of cell survival. CONCLUSION: Following massive small bowel resection, EGF retards rates of enterocyte apoptosis and modifies the expression of bcl-2 family members. By decreasing bax and increasing bcl-w expression, the balance between pro- and anti apoptotic genes is shifted in favor of cell survival. Alteration of bcl-2 family member expression may be an important mechanism by which EGF reduces the increased enterocyte apoptosis that occurs after massive small bowel resection. PMID- 10816348 TI - Extracellular matrix regulates the hepatocellular heat shock response. AB - BACKGROUND: Cirrhosis is characterized by the accumulation of collagen within the extracellular matrix (ECM) of the liver and progressive hepatocellular dysfunction. Since recent studies have shown that the ECM can modulate cellular function, we examined whether the ECM could contribute to hepatocellular dysfunction. To address this question we examined hepatocyte behavior in two different ECM environments. MATERIALS AND METHODS: Primary rat hepatocytes were cultured as a monolayer on collagen or as multicellular aggregates (spheroids) within a laminin-rich ECM. Hepatocytes were then compared for viability, response to proinflammatory cytokines, and their capacity to activate a heat shock response and adopt a thermotolerant phenotype. In addition, we compared the ability of prior heat shock exposure to protect hepatocytes from tumor necrosis factor (TNF) alpha/actinomycin-D-induced apoptosis in the two different ECM environments. RESULTS: Hepatocytes cultured as a monolayer on collagen exhibited decreased viability, underwent spontaneous apoptosis, and displayed an attenuated cytokine-stimulated nitric oxide production compared to hepatocytes cultured as spheroids. In response to heat, hepatocytes in both ECM environments expressed inducible heat shock protein 70 (hsp72). But, only the hepatocyte spheroids exhibited thermotolerance in response to a subsequent thermal challenge. In contrast to previous reports, induction of the heat shock response failed to protect hepatocytes against TNFalpha-induced apoptosis. CONCLUSIONS: These data demonstrate that the ECM can play an integral role in specific hepatocellular behaviors. Furthermore, the progressive deposition of collagen within the ECM, which is characteristic of fibrotic liver diseases, may directly contribute to the progressive hepatocellular dysfunction observed in cirrhosis. Hepatocellular viability, response to proinflammatory cytokines, heat shock response, and thermotolerance were all altered depending on the composition of the ECM. In contrast, TNFalpha-induced apoptosis was independent of the composition of the ECM. PMID- 10816349 TI - Extracorporeal circulation exacerbates microvascular permeability after endotoxemia. AB - BACKGROUND: Extracorporeal life support without prior inflammatory stimuli results in a modest increase in microvascular permeability. Initiating ECLS after shock, sepsis, or hypoxia results in marked increases in interstitial fluid and total body water. We sought to determine whether an inflammatory stimulus prior to initiating ECLS increases microvascular permeability to protein. METHODS: An anesthetized canine lymphatic fistula model was used to study Starling forces. LPS + ECLS received 1 mg/kg LPS 1 h prior to initiating ECLS, and the group ECLS received saline vehicle. To determine mesenteric microvascular permeability, mesenteric venous pressure was elevated to 32 +/- 1 mm Hg to reach a minimal lymph protein concentration (C(L)). With simultaneous measurement of plasma protein concentration (C(P)), the reflection coefficient, sigma, was calculated using the formula sigma = 1 - C(L)/C(P). Transvascular protein clearance and filtration coefficient (K(f)) were calculated from the measured Starling variables. After a steady state was achieved, normothermic right atrial-to femoral artery ECLS was initiated and continued for 2 h and then discontinued. Measurements were continued for 30 min after ECLS was discontinued. Measurements were continued for 30 min after ECLS was discontinued. Within-group comparisons were made with ANOVA and Fisher's LSD, and between-group comparisons were made with Student's t tests where appropriate. RESULTS: The reflection coefficient (sigma) decreased significantly from 0.77 +/- 0.02 to 0.53 +/- 0.07 with LPS + ECLS and was lower compared to ECLS alone (0.77 +/- 0.02 to 0.65 +/- 0.03). Transvascular protein clearance increased to a greater extent in LPS + ECLS from 266 +/- 46 to 819 +/- 125 microl/min compared to ECLS alone (284 +/- 49 to 819 +/ 125 microl/min) (P = 0. 06). K(f) increased in both groups after initiation of ECLS, but to a significantly greater extent in LPS + ECLS at 60 min (30.7 +/- 2.2 microl/min/mm Hg/g in ECLS and 50.0 +/- 8.9 microl/min/mm Hg/g in LPS + ECLS). Ileal tissue water increased in both groups, but there were no differences between groups. CONCLUSIONS: Initiation of ECLS after exposure to LPS increases ileal microvascular permeability to protein and water to a greater extent than ECLS alone. PMID- 10816350 TI - Bolus transit assessed by an esophageal stress test in postfundoplication dysphagia. AB - INTRODUCTION: Dysphagia is common after Nissen fundoplication but the relationship between dysphagia and bolus transit is poorly defined. This study compared bolus transit of fundoplication patients to normal individuals. METHODS: Twelve fundoplication patients and 20 healthy volunteers rated their ability to swallow eight bolus consistencies from no difficulty (0) to extreme difficulty (3) to compute a dysphagia score (range = 0-24). A 16-lumen manometric assembly was positioned across the esophagogastric junction (EGJ) and subjects were imaged fluoroscopically in a supine posture while swallowing 5 cc liquid barium and a 5 cc marshmallow-like viscoelastic barium bolus. Videofluoroscopic images were analyzed for total esophageal transit time and the fraction of time required to cross the EGJ. Manometric tracings were analyzed for the intrabolus pressure proximal to the EGJ, intragastric pressure, and distal peristaltic amplitude for each bolus. RESULTS: Dysphagia scores for fundoplication patients were significantly higher (7.3 +/- 5.1, range = 1-17) than for normals (0.5 +/- 0.6, range = 0-2). This correlated with longer total transit times for liquids and solids (r = 0.60, P < 0.01) and a greater percentage of transit time attributable to the EGJ transit. Retrograde flow at the EGJ (escape of bolus proximally up the esophagus) and peristaltic dysfunction were more frequent in fundoplication patients. However, no differences existed in manometric parameters between groups. CONCLUSIONS: Fundoplication impairs both liquid and solid esophageal bolus transit. Dysphagia perceived by fundoplication patients correlated with increased transit time, particularly across the EGJ. Combined quantitative evaluation with manometry and fluoroscopy reveals functional defects in fundoplication subjects, which are not evident by either modality alone. PMID- 10816351 TI - Hyperventilation increases muscle protein synthesis in critically ill trauma patients. AB - BACKGROUND: Critically ill trauma patients are often in negative nitrogen balance and demonstrate advanced muscle protein wasting, which is in part due to a decrease in muscle protein synthesis. Previous studies have been performed on the relationship between pH and protein metabolism. Some evidence suggests that alkalosis might enhance protein synthesis. The purpose of the present study is to determine whether protein synthesis is increased in trauma patients who have a respiratory alkalosis from hyperventilation. METHODS: Trauma patients in the intensive care unit (n = 8) who were treated with hyperventilation for elevated intracranial pressures were enrolled. Muscle protein synthesis rates were determined in vivo using the flooding method with l-[(2)H(5)]phenylalanine. Measurements were performed twice on each patient within a 36-h period, first during hyperventilation and then after hyperventilation was discontinued. Hemoglobin oxygen saturation was maintained above 95% for all measurements. RESULTS: Protein synthesis in muscle was 1.38 +/- 0.11%/day during hyperventilation (pH 7.50 +/- 0.02, pCO(2) 27.3 +/- 1.0 mm Hg) and 0.93 +/- 0.15%/day after respiratory parameters were normalized (pH 7.39 +/- 0.01, pCO(2) 39.4 +/- 1.5 mm Hg). The synthesis rate was significantly higher (P < 0.01, paired t test), 0.46 +/- 0.13%/day (32.6%), at the time of hyperventilation. CONCLUSION: Muscle protein synthesis is elevated during hyperventilation in critically ill patients with traumatic brain injury. We believe this preliminary study provides data that warrant confirmation in larger clinical studies. It suggests that this ventilatory therapeutic strategy may have a role in mitigating the negative nitrogen balance and muscle protein wasting that can impair the recovery of these patients. PMID- 10816352 TI - Focal adhesion kinase up-regulation and signaling in activated keratinocytes. AB - BACKGROUND: During wound healing keratinocytes undergo a process called "activation" that enables the cells to spread and migrate on wound matrix molecules. Focal adhesion kinase (FAK) is a key component of integrin-mediated intracellular signaling. We investigated the induction of FAK and its signaling activity during keratinocyte activation. MATERIALS AND METHODS: Keratinocytes were harvested from normal human skin. Previous work has shown that culture of keratinocytes causes activation in a manner similar to reepithelialization. Freshly isolated, unactivated cells were compared with cultured, activated cells. Activated cells were further examined either as growing colonies or after replating on type I collagen. FAK content was assessed by Western blotting. FAK distribution was shown using indirect immunofluorescence. FAK signaling activity was assessed using an antiphosphotyrosine antibody. RESULTS: FAK was not detectable by Western blotting in freshly isolated cells. In contrast FAK was detected in activated cells. FAK was up-regulated between Days 2 and 4 after cell isolation from skin. Immunostaining of activated, growing keratinocyte colonies in vitro showed a diffuse, cytoplasmic pattern. When these cells were replated on collagen, FAK became concentrated in focal adhesions. Lysates from replated cells showed increased tyrosine phosphorylation of FAK. CONCLUSIONS: In summary FAK is induced in keratinocytes in a time course comparable to that of activation. FAK is phosphorylated and undergoes redistribution to focal adhesions when cells are plated on the beta(1) integrin ligand collagen. These data suggest that induction of FAK and subsequent FAK-induced signaling may be responsible for changes in integrin-mediated behavior of activated keratinocytes during reepithelialization. PMID- 10816353 TI - Mechanism of the beneficial effects of pentoxifylline during sepsis: maintenance of adrenomedullin responsiveness and downregulation of proinflammatory cytokines. AB - BACKGROUND: Although it is known that pentoxifylline (PTX) produces various beneficial effects during sepsis, it remains unknown whether this agent has any salutary effects on the depressed vascular responsiveness to adrenomedullin (ADM), a novel potent vasodilatory peptide, under such conditions. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). One hour after CLP, PTX (50 mg/kg body wt) or vehicle (normal saline) was infused intravenously over 90 min. Twenty hours after CLP (i.e., the late, hypodynamic stage of sepsis), the thoracic aorta and small intestine were isolated and preconstricted by norepinephrine. Rat ADM (10(-7) M) was applied, and the percentage of ADM-induced relaxation in the aortic rings and resistance vessels in the small intestine was determined. In addition, plasma ADM was determined by radioimmunoassay and tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6 levels were measured by enzyme-linked immunosorbent assay. RESULTS: The percentage of ADM-induced vascular relaxation in the aortic rings and resistance vessels of the isolated gut was significantly reduced 20 h after CLP. Administration of PTX early after the onset of sepsis, however, prevented the decrease in vascular ADM responsiveness at the macro- and microcirculatory levels. Plasma ADM levels increased after CLP, irrespective of PTX infusion, indicating that the effect of PTX was not mediated by altering ADM release. The upregulated TNF-alpha, IL-1beta, and IL-6 during late sepsis were, however, attenuated by PTX administration, suggesting that maintenance of ADM responsiveness by this agent appears to be due to downregulation of these cytokines. CONCLUSIONS: Since early administration of PTX maintains vascular ADM responsiveness even during the late stage of sepsis, this agent appears to be a useful adjunct in preventing the deterioration in hemodynamics and cardiovascular function during the progression of polymicrobial sepsis. PMID- 10816354 TI - Glucagon-like peptide 2: a new treatment for chemotherapy-induced enteritis. AB - BACKGROUND: Glucagon-like peptide 2 (GLP-2) is a recently identified intestinal epithelium-specific growth factor that has been shown to reduce the severity of inflammatory disorders of the intestine in rodent models. We hypothesized that GLP-2 administration would be beneficial in chemotherapy-induced enteritis either by preventing injury or by promoting recovery. MATERIAL AND METHODS: Rats received no drug (control), chemotherapy alone [5-fluorouracil (5-FU), 190 mg/kg, ip] (Chemo), 5-FU followed by 3 days of GLP-2 analog (ALX-0600, 0.1 microg, sc twice daily) (CH-G), or GLP-2 analog for 6 days prior to 5-FU and for 3 days afterward (G-CH-G). Animals were pair fed. Rats received 5-bromo-2-deoxyuridine (Br-dU, 50 mg/kg, 2.5 h prior to sacrifice on Day 3 postchemotherapy) for immunohistochemical assessment of cellular proliferation. RESULTS: Chemotherapy induced significant reductions in body weight, villus height, and crypt depth compared with controls. Intestinal wet weight, villus height, and crypt depth were significantly higher for the CH-G group compared with the Chemo group. The CH-G group also showed a significant improvement in villus height compared with the G-CH-G group. Crypt depth, but not jejunal wet weight or villus height, was significantly improved in the G-CH-G group compared with the Chemo group. The percentage of Br-dU-labeled cells in the intestinal crypts did not differ among the groups. CONCLUSIONS: These results suggest, for the first time, that GLP-2 treatment initiated after chemotherapy administration enhances intestinal recovery. In contrast, GLP-2 treatment initiated prior to chemotherapy administration to prevent injury has less beneficial effect. GLP-2 administration may be beneficial to patients suffering from chemotherapy-induced enteritis. PMID- 10816355 TI - The effect of age and prolonged cold ischemia times on the national allocation of cadaveric renal allografts. AB - BACKGROUND: National sharing of cadaveric renal allografts for perfectly matched kidneys (0 antigen mismatch) has improved outcome in the recipients of these kidneys despite increasing cold storage times. However, there may be limits to outcome improvement of matched kidneys based on age and cold storage time. MATERIALS AND METHODS: To determine if national sharing of kidneys based on matching improves outcome regardless of donor age and cold storage time, we evaluated the United Network for Organ Sharing (UNOS) Scientific Registry for all recipients of cadaveric kidney transplants between January 1, 1990 and July 31, 1998. We divided the recipients into four groups based on donor age and cold storage time. Group 1 comprised young donors (donor age <55 years) with average (<24 h) cold storage time; group 2, young donors with long (>/=24 h) cold storage time; group 3, older donors (donor age >/=55 years) with average cold storage time; and group 4, older donors with long cold storage time. RESULTS: A total of 64,046 recipients were evaluated: 35,061 (55%) in group 1, 21,264 (33%) in group 2, 4308 (7%) in group 3, and 3414 (5%) in group 4. Early graft performance progressively decreased from group 1 to group 4. Delayed graft function (DGF: dialysis requirement in the first 7 days posttransplant) was 18, 29, 33, and 42% (P < 0.0001); serum creatinine at 3 years (in mg/dl) was 1.70 +/- 0.8, 1.73 +/- 0.9, 2. 31 +/- 1.0, and 2.42 +/- 1.1 (P < 0.0001); 1-year graft survival was 87, 84, 79, and 77% (P < 0.0001); and 3-year graft survival was 77, 74, 63, and 62% (P < 0.0001, for groups 1 and 2 vs groups 3 and 4, respectively). The trends in DGF persisted through the groups in 0 antigen mismatched kidneys. CONCLUSIONS: Early function is adversely affected by prolonged cold storage, despite matching, in recipients of younger and older donor kidneys. Long-term function does not appear to be affected by prolonged cold storage. Recipients of kidneys from donors >/=55 years of age have significantly worse short- and long-term outcome and may not benefit from national sharing. PMID- 10816356 TI - Outcome and quality of life of patients with acute pancreatitis requiring intensive care. AB - BACKGROUND: Patients with severe acute pancreatitis often require intensive care unit (ICU) admission, have multiple complications, spend weeks to months in the hospital, and consume a large amount of resources. The aim of this study was to evaluate the ICU course, costs, mortality, and quality of life of patients who require ICU admission for acute pancreatitis. METHODS: Patients with acute pancreatitis requiring ICU admission were identified retrospectively. Data regarding in-hospital morbidity, mortality, and hospital costs were obtained. Long-term quality of life was assessed using the Short Form-36 Health Survey (SF 36). RESULTS: Fifty-two patients were identified. There were 31 men and 21 women: the mean age was 53 years (range, 22-89). The most common causes of acute pancreatitis were gallstones (44%) and alcoholism (17%). Pulmonary failure (52% required mechanical ventilation) and renal failure (21% required dialysis) were common. There were 39 (75%) hospital survivors and 13 (25%) nonsurvivors. In the first 24 h, the mean Acute Physiology and Chronic Health Evaluation (APACHE) II scores were 10 +/- 6 in survivors and 16 +/- 4 in the nonsurvivors (<0.01). Mean length of ICU (15 +/- 18 and 28 +/- 31 days) and hospital (40 +/- 34 and 38 +/- 34 days) stays were similar in survivors and nonsurvivors, respectively (NS). The mean hospital cost for survivors was $83,611 +/- 88,434 and that for nonsurvivors was $136,730 +/- 95,045 (P = 0. 09). The estimated cost to obtain one hospital survivor was $129,188. Of the 39 hospital survivors, 5 died later, 21 completed the SF-36, and 13 were lost to follow-up. Long-term quality of life (SF-36) was similar to that of an age-matched population. Twenty of twenty-one felt their general health was at least as good as it had been 1 year previously. CONCLUSIONS: Patients with severe acute pancreatitis need prolonged ICU and hospital stays. APACHE II may be a good predictor of outcome; further, prospective evaluation is needed. Although resource utilization is high, most patients survive and have good long-term quality of life. PMID- 10816357 TI - Genetic regulation of root hair development in Arabidopsis thaliana: a network model. AB - The root epidermis of Arabidopsis thaliana is formed by alternate files of hair and non-hair cells. Epidermal cells overlying two cortex cells eventually develop a hair, while those overlying only one cortex cell do not. Here we propose a network model that integrates most of the available genetic and molecular data on the regulatory and signaling pathways underlying root epidermal differentiation. The network architecture includes two pathways; one formed by the genes TTG, R homolog, GL2 and CPC, and the other one by the signal transduction proteins ETR1 and CTR1. Both parallel pathways regulate the activity of AXR2 and RHD6, which in turn control the development of root hairs. The regulatory network was simulated as a dynamical system of eight discrete state variables. The distinction between epidermal cells contacting one or two cortical cells was accounted for by fixing the initial states of CPC and ETR1 proteins. The model allows for predictions of mutants and pharmacological effects because it includes the ethylene receptor. The dynamical system reaches one of the six stable states depending upon the initial state of the CPC variable and the ethylene receptor. Two of the stable states describe the activation patterns observed in mature trichoblasts (hair cells) and atrichoblasts (non-hair cells) in the wild-type phenotype and under normal ethylene availability. The other four states correspond to changes in the number of hair cells due to experimentally induced changes in ethylene availability. This model provides a hypothesis on the interactions among genes that encode transcription factors that regulate root hair development and the proteins involved in the ethylene transduction pathway. This is the first effort to use a dynamical system to understand the complex genetic regulatory interactions that rule Arabidopsis primary root development. The advantages of this type of models over static schematic representations are discussed. PMID- 10816358 TI - Comparing risk factors for population extinction. AB - Extinction risk of natural populations of animals and plants is enhanced by many different processes, including habitat size reduction and toxic chemical exposure. We develop a method to evaluate different risk factors in terms of the decrease in the mean extinction time. We choose a population model with logistic growth, environmental and demographic stochasticities with three parameters (intrinsic growth rate r, carrying capacity K, and environmental noise sigma(2)(e)). The reduction in the habitat size decreases carrying capacity K only, whilst toxic chemical exposure decreases survivorship (or fertility) and in effect reduces both r and K. We derived a formula for the reduction in habitat size that decrease the mean extinction time by the same magnitude as a given level of toxic chemical exposure. In a large population (large K) or in a slowly growing population (small r), a small decrease in survivorship can cause the extinction risk increase corresponding to a significant reduction in the habitat size. This conclusion depends also on the nonlinearity of dose-effect relationship. To illustrate the method, we analyse a freshwater fish, Japanese crucian carp (Carassius auratus subsp.) in Lake Biwa. PMID- 10816359 TI - Extinction risk of a density-dependent population estimated from a time series of population size. AB - Environmental threats, such as habitat size reduction or environmental pollution, may not cause immediate extinction of a population but shorten the expected time to extinction. We develop a method to estimate the mean time to extinction for a density-dependent population with environmental fluctuation. We first derive a formula for a stochastic differential equation model (canonical model) of a population with logistic growth with environmental and demographic stochasticities. We then study an approximate maximum likelihood (AML) estimate of three parameters (intrinsic growth rate r, carrying capacity K, and environmental stochasticity sigma(2)(e)) from a time series of population size. The AML estimate of r has a significant bias, but by adopting the Monte Carlo method, we can remove the bias very effectively (bias-corrected estimate). We can also determine the confidence interval of the parameter based on the Monte Carlo method. If the length of the time series is moderately long (with 40-50 data points), parameter estimation with the Monte Carlo sampling bias correction has a relatively small variance. However, if the time series is short (less than or equal to 10 data points), the estimate has a large variance and is not reliable. If we know the intrinsic growth rate r, however, the estimate of K and sigma(2)(e)and the mean extinction time T are reliable even if only a short time series is available. We illustrate the method using data for a freshwater fish, Japanese crucian carp (Carassius auratus subsp.) in Lake Biwa, in which the growth rate and environmental noise of crucian carp are estimated using fishery records. PMID- 10816360 TI - Modeling size-dependent photosynthesis: light absorption and the allometric rule. AB - Microalgal photosynthesis can be predicted using empirical allometric or mechanistic bio-optic models. These two descriptions are usually considered independently. We compare the size scaling of photosynthesis predicted by these two models. Size scaling exponents for phytoplankton often deviate from the allometric 3/4 rule. This may be because the allometric model does not account for the size dependence of light absorption and its effect on the size scaling of photosynthesis. In contrast to the allometric model and experimental data, the bio-optic model predicts photosynthesis should be independent of cell size when intracellular pigment concentrations are low or inversely related to cell diameter. A composite of the allometric and bio-optic models is described and compared to laboratory data of light-limited nutrient-saturated diatom photosynthesis. The allo-bio-optic model provides a mechanistic explanation for the anomalous size scaling found in laboratory and field studies of microalgal photosynthesis and growth. PMID- 10816361 TI - Impaired vascular dynamics in normotensive diabetic rats induced by streptozotocin: tapered T-tube model analysis. AB - This study is to explore the changes of arterial mechanical properties in streptozotocin (STZ)-diabetic rats, based on the exponentially tapered T-tube model. Rats given STZ 65 mg kg(-1)i.v. are compared with untreated weight- and age-matched controls. A high-fidelity pressure sensor and electromagnetic flow probe measured pulsatile pressure and flow waves in the ascending aorta, respectively. Diabetic rats exhibit isobaric vasodilatation that is characterized by an increase in cardiac output and no significant changes in aortic pressure. Total peripheral resistance of diabetic rats is lower than that of weight- and age-matched controls. Diabetic rats have higher total peripheral compliance (2.86+/-0.70 microl mm Hg(-1)) than do weight- (1.77+/-0.34 microl mm Hg(-1)) and age-matched (1.87+/-0.69 microl mm Hg(-1)) controls. Aortic characteristic impedance is reduced from 0.017+/-0.003 mm Hg min kg ml(-1)in weight- and 0.020+/ 0.004 mm Hg min kg ml(-1)in age-matched controls to 0.010+/-0.004 mm Hg min kg ml(-1)in diabetic rats. Moreover, diabetic rats show shorter wave transit time in lower body circulation (17.86+/-1.91 ms) than do weight- (20.45+/-1.91) and age matched (23.05+/-2.04 ms) controls. Under isobaric vasodilatation, the decreased resistance and increased compliance in peripheral circulation suggest that the contractile dysfunction of the smooth muscle cells may occur in resistance arterioles in diabetes. With unaltered aortic pressure, an impairment in aortic distensibility of STZ-diabetic rats is manifest on the reduced wave transit time rather than on the diminished aortic characteristic impedance. PMID- 10816363 TI - Modeling of diffusion and concurrent metabolism in cutaneous tissue. AB - Clearance by cutaneous metabolism can shield the body from penetration of environmental and therapeutic xenobiotics. Here we report on a physical model to relate Fickian diffusion and concurrent Michaelis-Menten metabolism of drugs in the viable epidermis of human skin. For this purpose, we numerically generated substrate concentration profiles within the metabolizing tissue and the resulting donor-to-receiver substrate fluxes through the tissue for various mass transport and metabolism parameters. To validate the model, permeation and concurrent metabolism of a peptidomimetic compound, L -Ala-4-methoxy-2-naphthylamide (Ala MNA), across both stripped human skin and HaCaT cell culture sheets were compared to numerical simulations. Parameter estimates for those calculations were extracted from independent experiments. Experimental data and numerical predictions were in excellent agreement. Also, numerical fits and independently validated parameters correlated closely, indicating the principal validity of the physical model. Numerical simulations and theoretical derivations illustrate the kinetic impact of the factors involved, i.e. the diffusion coefficient D, substrate donor concentration C(S,D), substrate partition coefficient P, tissue thickness L and maximum metabolic rate V(max), on drug permeation, with L having the strongest effect. In the steady state, the coefficient 2 alpha, i.e. the dimensionless ratio of the residence time term (L(2)/D) of a substrate in the tissue to the metabolic half-life term (C(S,D)P/2 V(max)), allows to estimate concentration gradients within the tissue and the extent of metabolism. High 2 alpha values represent practically complete metabolic cleavage upon penetration. Epidermis ( approximately 40 microm thick) of stripped human skin and HaCaT sheets ( approximately 10 microm) had 2 alpha values of 43 and 2.7, respectively, indicating that intact Ala-MNA could only permeate HaCaT sheets, but not skin. Independent permeation experiments confirmed this outcome. This physical model may be applicable to other metabolizing tissues as well. PMID- 10816362 TI - A probabilistic model for ligand-cytoskeleton transmembrane adhesion: predicting the behavior of microspheres on the surface of migrating cells. AB - A theoretical model describing the attachment and cytoskeletal coupling of microspheres to the dorsal surface of motile cells was developed. Integral membrane receptors beneath a ligand-coated microsphere are allowed to be either free, attached to the microsphere, bound to the rearward moving actin network, or linked to both the bead and the cytoskeleton, and to switch between these four states. The binding transitions being modeled as chemical reactions governed by rate constants taken from literature, the chance for a receptor to be in each binding state over time is obtained by solving mass-balance equations for the probability functions. The population of n such receptors beneath the microsphere is accounted for by a binomial distribution for each state. Adhesion and transmembrane coupling (resulting in microsphere transport) being defined by a minimal number of ligand-receptor and receptor-cytoskeleton bonds, respectively, the probabilities of attachment and transport of the microsphere over time are expressed in terms of state probability distributions. It is found that increasing the ligand density raises the attachment and transport probabilities, in good quantitative agreement with recent experiments using optical tweezers and accurate position tracking. Increasing the bead size does not affect attachment, but raises the transport probability with a marked transition for bead diameter around 100 nm, as for experimental data. Increasing the restraining force decreases the transport probability, probably by inducing a rupture of receptor cytoskeleton bonds. This study thus provides a framework that helps understand the process of cortical flow associated with cell locomotion. PMID- 10816364 TI - Learning and the theory of games. AB - The potential for Game Theory in the Practice of Psychiatry is outlined. The deficiences of current Learning Theory in explaining observed phenomena are highlighted and the potential for a Game Theory analysis highlighted. The extension to "Game Learning" is of broad application. PMID- 10816365 TI - Optimal trajectories for the short-distance foraging flights of swans. AB - Optimal flight theory relates body measurements (wing span, body cross-section, body mass) and aerodynamic variables (air density, drag, profile and induced power ratios) to the most energy-efficient velocity for long distance migration. For short-range (2-10 km) foraging flights the theory is expanded to include non negligible costs for take-off and energy savings/losses for climbing to altitude (drag decreases with air density and therefore with altitude). The theory predicts clear differences between Tundra and Trumpeter swans. Generally speaking, for flights between 2 and 10 km Trumpeter swans can be expected to fly approximately 5-10 m lower in altitude and 1-2 ms(-1)more slowly than Tundra swans. Moreover, the total energy required for these foraging flights is approximately 150% larger for a Trumpeter than a Tundra swan (80 vs. 120 kJ of direct mechanical energy for a 5 km flight), suggesting that Trumpeter swans may be less inclined to take-off than Tundra swans. These factors indicate that even Trumpeters native to the area (as opposed to recently translocated) would be more vulnerable to hunting than native Tundra swans. The expanded theory is compared to observations made in Utah's Bear River Migratory Bird Refuge. PMID- 10816366 TI - Competitive coexistence in antiviral immunity. AB - Adaptive immunity to viruses in vertebrates is mediated by two distinct but complementary branches of the immune system: the cellular response, which eliminates infected cells, and the humoral response, which eliminates infectious virus. This leads to an interesting contest, since the two responses compete, albeit indirectly, for proliferative stimuli. How can a host mount a coordinated antiviral campaign? Here we show that competition may lead to a state of "competitive coexistence" in which, counterintuitively, each branch complements the other, with clinical benefit to the host. The principle is similar to free market economics, in which firms compete, but the consumer benefits. Experimental evidence suggests this is a useful paradigm in antiviral immunity. PMID- 10816367 TI - Haldane's rule: hybrid sterility affects the heterogametic sex first because sexual differentiation is on the path to species differentiation. AB - Prevention of recombination is needed to preserve both phenotypic differentiation between species and sexual phenotypic differentiation within species. For species differentiation (speciation), isolating barriers preventing recombination may be pre-zygotic (gamete transfer barriers), or post-zygotic (either a developmental barrier resulting in hybrid inviability, or a chromosomal-pairing barrier resulting in hybrid sterility). The sterility barrier is usually the first to appear and, although often initially only manifest in the heterogametic sex (Haldane's rule), is finally manifest in both sexes. For sexual differentiation, the first and only barrier is chromosomal-pairing, and always applies to the heterogametic sex. For regions of sex chromosomes affecting sexual differentiation there must be something analogous to the process generating the hybrid sterility seen when allied species cross. Explanations for Haldane's rule have generally assumed that the chromosomal-pairing barrier initiating evolutionary divergence into species is due to incompatibilities between gene products ("genic), or sets of gene products ("polygenic), rather than between chromosomes per se ("chromosomal"). However, if chromosomal incompatibilities promoting incipient sexual differentiation could also contribute to the process of incipient speciation, then a step towards speciation would have been taken in the heterogametic sex. Thus, incipient speciation, manifest as hybrid sterility when "varieties" are crossed, would appear at the earliest stage in the heterogametic sex, even in genera with homomorphic sex chromosomes (Haldane's rule for hybrid sterility). In contrast, it has been proposed that Haldane's rule for hybrid inviability needs differences in dosage compensation, so could not apply to genera with homomorphic sex chromosomes. PMID- 10816368 TI - A hypercycle theory of proliferation of viruses and resistance to the viruses of transgenic plant. AB - A set of dynamical equations for the proliferation of two typical viruses TMV and PVY has been derived from the reaction equations describing their replication, assembly and translation. These equations can be seen as the generalization of hypercycle theory to the system. The quantitative explanation on the phenomena of proliferation of plant virus and the mechanism of resistance to the disease of transgenic plant is offered. The phenomenon of specific cessation of minus-strand RNA accumulation in the proliferation of TMV, the cross-protection of plant viruses and the mechanism of resistance to viruses of transgenic plant are discussed based on the computer simulation of the proliferation of viruses and the prediction of the secondary structure of the genomic RNA. PMID- 10816369 TI - Competitive speciation in quantitative genetic models. AB - We study sympatric speciation due to competition in an environment with a broad distribution of resources. We assume that the trait under selection is a quantitative trait, and that mating is assortative with respect to this trait. Our model alternates selection according to Lotka-Volterra-type competition equations, with reproduction using the ideas of quantitative genetics. The recurrence relations defined by these equations are studied numerically and analytically. We find that when a population enters a new environment, with a broad distribution of unexploited food sources, the population distribution broadens under a variety of conditions, with peaks at the edge of the distribution indicating the formation of subpopulations. After a long enough time period, the population can split into several subpopulations with little gene flow between them. PMID- 10816370 TI - The interaction of pathogens with humans. PMID- 10816372 TI - Expression and regulation of the streptokinase gene. AB - Recent research in various areas has appreciably expanded our knowledge of streptokinase, a plasminogen activator produced by all human group A (GAS), group C (GCS), and group G (GGS) streptococci. Several molecular genetic approaches are described here to study the expression of the streptokinase gene, skn. Southern hybridization analysis demonstrated homology of synteny of ska, skc, and skg in the genomes of the above serogroups. S1 nuclease mapping, the use of transcriptional fusions to beta-galactosidase and luciferase reporter genes, in conjunction with site-directed mutagenesis, led to the localization of the core promoter region of skc and the identification of a cis-active upstream region required for full promoter activity. Circular permutation analysis of the promoter upstream region identified an intrinsic DNA bending locus as the pivotal DNA element stimulating the activity of the core promoter. The detection of skn allele-specific expression phenotypes, which proved not to be due to different skn mRNA half-lives, prompted allele swap experiments, showing that promoter activity is dictated by the host genetic background, rather than the sequence of the regulatory region. These findings suggest the involvement in skn expression of an as yet unidentified transcriptional activator that contacts the bent DNA region. Transcription termination of skc is directed by a bidirectional terminator whose structural requirements for termination efficiency were determined with base substitution mutants fused to a chloramphenicol acetyl transferase reporter. Finally, mutagenic plasmids are described for insertion duplication and allele replacement mutagenesis of the skn locus. PMID- 10816371 TI - Nonfibrinolytic functions of plasminogen. AB - Although the roles of plasminogen and plasmin in mediating blood clot dissolution are well known, the availability of mice deficient for components of the fibrinolytic system has allowed direct approaches to be made toward elucidating the role of these proteins in other diverse physiological and pathophysiological processes. A number of these studies have identified plasminogen as playing an important role in inflammation and other cell migratory processes. With the identification of receptors for plasminogen on a number of pathogens, and the ability to activate plasminogen through either endogenous production of plasminogen activators or utilization of host activators, mice deficient for components of the fibrinolytic system offer a unique approach toward further elucidating the importance of this system in pathogen infection and dissemination. PMID- 10816373 TI - Plasminogen activation in degradation and penetration of extracellular matrices and basement membranes by invasive bacteria. AB - Methods to assess in vitro the role of plasminogen activation in enterobacterial degradation of extracellular matrices and their protein components as well as in penetration through basement membrane are described. Development of these methods was initiated after the findings that enterobacterial surface structures (fimbriae and the Pla surface protease) function in plasminogen activation as well as in laminin- and/or fibronectin-specific adhesion. Enterobacteria with these properties degrade radiolabeled laminin as well as metabolically labeled extracellular matrix from cultured endothelial or epithelial cells. Plasmin coated bacteria also penetrate through the reconstituted basement membrane preparation Matrigel. The processes are dependent on plasminogen activation by the invasive bacteria. The results suggest a pathogenic similarity between enterobacteria and tumor cells in cellular metastasis through tissue barriers. PMID- 10816374 TI - The generation of enzymatically active plasmin on the surface of spirochetes. AB - The spirochete Borrelia burgdorferi, the etiologic agent of Lyme disease, is transmitted to the host by a feeding Ixodid tick. The spirochete subsequently disseminates through the skin, enters the bloodstream, and becomes systemic. A potential mechanism for this invasiveness was identified with the discovery that B. burgdorferi can bind components of the plasminogen activation system (PAS). The methodology for analyzing the generation of enzymatically active plasmin on the surface of this organism is given, and applied to measure spirochete viability, strain differences, and breakdown of extracellular matrix (ECM) macromolecules. Plasmin acquisition by B. burgdorferi was measured photometrically by a specific chromogenic substrate. The growth of B. burgdorferi in culture was not affected by the presence of active plasmin on the spirochete surface. Plasmin-coated B. burgdorferi degraded the purified (ECM) components fibronectin, laminin, and vitronectin, but not collagen. The addition of B. burgdorferi with surface plasmin to a radiolabeled, native ECM resulted in degradation of noncollagenous protein, as measured by release of solubilized radioactivity. Breakdown of purified ECM components or native ECM did not occur after exposure to untreated spirochetes or spirochetes treated with uPA or PLG alone. These results provide in vitro evidence that enzymatically active plasmin on the surface of B. burgdorferi may be partially responsible for its invasiveness. PMID- 10816375 TI - Analysis of plasminogen-binding M proteins of Streptococcus pyogenes. AB - Group A streptococci are common human pathogens that cause a variety of infections. They express M proteins which are important cell wall-bound type specific virulence factors. We have found that a set of strains, associated primarily with skin infections, express M proteins that bind plasminogen and plasmin with high affinity. The binding is mediated by a 13-amino-acid internal repeated sequence located in the N-terminal surface-exposed portion of these M proteins. This sequence binds to kringle 2 in plasminogen, a domain that is not involved in the interaction with streptokinase, a potent group A streptococcal activator of plasminogen. It could be demonstrated that plasminogen, absorbed from plasma by growing group A streptococci expressing the plasminogen-binding M proteins, could be activated by exogenous and endogenous streptokinase, thereby providing the bacteria with a surface-associated enzyme that could act on the tissue barriers in the infected host. PMID- 10816376 TI - Helicobacter pylori interactions with plasminogen. AB - Helicobacter pylori is the causative agent of chronic gastritis, peptic ulcer, and gastric malignancies. A number of virulence factors have been described including several adhesins, a cytotoxin, neutrophil-activating protein, and expression of binding of extracellular matrix proteins, like collagen type IV, laminin, and vitronectin. H. pylori strains commonly express binding of soluble plasminogen. Coccoid forms also express binding. Plasminogen binding was optimal at pH 7.0. The binding is mediated by two cell surface proteins of 42 and 57 kDa. Scatchard plot analysis showed a straight line with a K(d) of 7 x 10(-7) M. Lysine and E-aminocaproic acid inhibited binding. The binding domain on the plasminogen molecule is the fifth kringle, miniplasminogen. Plasminogen is converted to plasmin by tissue plasminogen activator. During H. pylori infection the activity of tissue plasminogen activator is decreased and that of urokinase increased. This is reversed after eradication therapy. The plasminogen binding and conversion to plasmin is the only proteolytic activity of H. pylori, and may enhance tissue penetration and be involved in carcinogenesis. PMID- 10816377 TI - Assays for functional binding of plasminogen to viral proteins. AB - Cleavage of the hemagglutinin (HA) molecule by proteases is a prerequisite for the infectivity of influenza A viruses. Plasminogen binds to the viral glycoprotein neuraminidase (NA), and NA-bound plasminogen is activated to plasmin, which cleaves the HA of influenza A/WSN/33 (WSN) (H1N1) virus. Here we present assays for detecting functional plasminogen binding to the influenza virus NA. PMID- 10816378 TI - Interaction of group A streptococci with human plasmin(ogen) under physiological conditions. AB - A series of methods for analyzing the interaction of group A streptococci with the human plasminogen system are described. Examples of group A streptococcal isolates capable of assembling surface plasminogen activator activity when grown in human plasma are presented and the key requirements for this process are evaluated. The stabilities of cell-associated plasmin and plasminogen activator complexes are compared and a model for the interaction of group A streptococci with the plasminogen system in an infected host is presented. PMID- 10816379 TI - Role of the pleiotropic effects of plasminogen deficiency in infection experiments with plasminogen-deficient mice. AB - Plasminogen-deficient mice hold great promise as tools for analyzing the contribution of plasminogen activators produced by infectious agents to pathogenesis. However, the pathology caused by congenital plasminogen deficiency complicates the interpretation of infection experiments conducted with these animals. This pathology, the most prominent features of which are poor weight gain, wasting after about 60 days of age, and shortened lifespan, results from the inability of the mice to clear small fibrin thrombi. This article describes strategies for distinguishing the contribution of this pathology from the direct effects of depriving infectious agents of plasminogen. These strategies depend on the use of mouse genotypes in which the correlation of plasminogen deficiency with fibrin-dependent pathology is broken. Mice with plasminogen activator deficiencies are unable to generate plasmin and develop pathologies identical to those seen in plasminogen-deficient mice. However, unlike plasminogen-deficient mice, they do make plasminogen available to the infectious agent. Fibrinogen deficient mice also deficient for plasminogen do not develop the pathology typical of plasminogen deficiency. These mice allow examination of plasminogen deficiency in the absence of fibrin-dependent pathology. Use of fibrinogen deficient mice is complicated by the possibility that fibrin may be the key substrate of plasmin generated by the infectious agent. PMID- 10816380 TI - The potential role for nephritis-associated plasmin receptor in acute poststreptococcal glomerulonephritis. AB - Immunoglobulin G from a patient convalescing from acute poststreptococcal glomerulonephritis (APSGN) bound specific antigenic sites in early APSGN glomeruli. A streptococcal cytoplasmic antigen (preabsorbing antigen, PA-Ag), could selectively preabsorb fluorescein isothiocyanate (FITC)-labeled IgG and prevented glomerular staining. The antigen was purified and identified as an M(r) approximately 43,000 protein with a pI of 4.7 that strongly activated complement C3 (N. Yoshizawa, S. Oshima, I. Sagel, J. Shimizu, and G. Treser, 1992, J. Immunol. 148, 3110-3116). In the present study, a nephritogenic antigen was purified by affinity chromatography using APSGN IgG-immobilized Sepharose followed by chromatography on an anion-exchange resin. Purification was monitored by ELISA and Western blotting using the binding characteristics of the specific antibodies present in APSGN serum. The molecular weight of the purified antigen, named nephritis-associated plasmin receptor (NAPlr), was an M(r) approximately 43,000 protein and the internal amino acid sequence was found to be homologous to those of the plasmin receptor (Plr) of group A streptococci strain 64/14 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus subtilis. The purified NAPlr exhibited GAPDH activity and plasmin(ogen) binding activity. Using FITC-labeled rabbit anti-NAPlr, the antigen was found to be present in the glomeruli of 22 of 22 patients in the early stage of APSGN. Bacterial Plr was also demonstrated in human APSGN glomeruli for the first time using monoclonal antibody to the recombinant Plr protein. Antibody to NAPlr was found in the sera of 46 of 50 (92%) patients within 3 months of onset. These results led us to speculate that NAPlr bound to the glomeruli may contribute to the pathogenesis of APSGN via plasmin and complement activation. PMID- 10816381 TI - Interactions of CCR5 and CXCR4 with CD4 and gp120 in human blood monocyte-derived dendritic cells. AB - Dendritic cells (DC) and macrophages play an important role in the generation of immune responses and transmission of HIV infection. It has been recently found that, in the presence of gp120, CD4 can be efficiently coimmunoprecipitated by anti-CXCR4 antibodies from lymphocytes and monocytes but not from blood monocyte derived macrophages. The gp120-CD4-CXCR4 complex formation paralleled the ability for these cell types to support X4 (LAV) HIV-1 envelope glycoprotein (Env) mediated fusion. Here we report that, unlike macrophages but similar to lymphocytes and monocytes, human blood monocyte-derived DC allow efficient complex formation among the HIV-1 coreceptor CXCR4, the primary receptor CD4, and the Env gp120 (LAV) which parallels their fusion ability with cells expressing HIV-1 Env (LAV). In addition, DC behaved similarly to macrophages, lymphocytes, and monocytes in their ability to support formation of complexes between CD4 and the other major HIV-1 coreceptor CCR5 even in the absence of gp120 as demonstrated by CD4 coimmunoprecipitation with anti-CCR5 antibodies. Further, the amount of gp120-CD4-CXCR4 (or CCR5) complexes was proportional to the extent of cell fusion mediated by the HIV-1 Env (LAV or JRFL, respectively). These results demonstrate that of all the major types of host cells important for HIV-1 infection, the first central stage in the entry mechanism, the formation of gp120 CD4-coreceptor complexes, is not impaired except for the formation of the gp120 CD4-CXCR4 complex in macrophages. Therefore, for most CD4+ target cells restraint(s) on productive HIV-1 infection appears to occur at stages of the virus life cycle subsequent to the gp120-CD4-coreceptor complex formation. PMID- 10816382 TI - Inefficient formation of a complex among CXCR4, CD4 and gp120 in U937 clones resistant to X4 gp120-gp41-mediated fusion. AB - Certain subclones (designated as minus clones) of the promonocytic U937 cell line do not support efficient infection and fusion mediated by T cell line adapted (TCLA) X4 HIV-1 gp120-gp41 (Env) although the CXCR4 and CD4 concentrations at their surfaces are similar to those at the surfaces of clones susceptible to HIV 1 entry (plus clones) (H. Moriuchi et al., J. Virol. 71, 9664-9671, 1997). To test the hypothesis that inefficient formation of gp120-CD4-CXCR4 complexes could contribute to the mechanism of resistance to Env-mediated fusion in the minus clones, we incubated plus and minus cells with HIV-1 LAI gp120 and coimmunoprecipitated CD4 by using anti-CXCR4 antibodies. The gp120 induced inefficient coimmunoprecipitation of CD4 in the minus clones but not in the plus ones. Overexpression of CD4 resulted in significant restoration of the minus clones' susceptibility to fusion in parallel with an increase in the amount of the gp120-CD4-CXCR4 complexes. These results not only suggest that the resistance to TCLA X4 HIV-1 entry in the U937 minus clones is due to the inability of these cells to efficiently form complexes among CD4, gp120, and CXCR4, but also provide a direct evidence for the correlation between fusion and the cell surface concentration of the complexes among CXCR4, CD4, and gp120. These data and similar recent observations in macrophages suggest that inefficient complex formation among CXCR4, CD4, and gp120 could be a general mechanism of cell resistance to gp120-gp41-mediated fusion and a major determinant of HIV-1 evolution in vivo. PMID- 10816383 TI - Spontaneously regressing oral papillomas induce systemic antibodies that neutralize canine oral papillomavirus. AB - Canine oral papillomavirus (COPV) infection of naive beagle dogs causes oral papillomas, most of which spontaneously regress. Regressor beagles do not develop new oral papillomas because of COPV type-specific, cell-mediated immunity, COPV neutralizing antibodies, or both. Formalin-fixed native and recombinant COPV vaccines that target the systemic immune system induce neutralizing antibodies that prevent development of oral papillomas. This study was designed to determine whether spontaneously regressing mucosal papillomas also targeted the systemic immune system to induce circulating, neutralizing IgG antibodies that protect against infection by COPV. To accomplish this goal, IgG was fractionated from sera collected from weanling beagles and regressor beagles and tested for conferring protection by passive immunization. Serum was tested by ELISA for antibodies against intact virions and then pooled for passive transfer to naive beagles. Preimmune sera were neither reactive by ELISA nor protective by passive transfer. On the other hand, IgG antibodies from regressor beagles were reactive by ELISA and passive transfer conferred protection against COPV challenge. Circulating IgG antibodies induced by spontaneous regression of canine oral papillomas protect beagles against intraoral infection by COPV, a model for mucosotropic HPV. PMID- 10816384 TI - Differentiation status of rat ductal cells and ethionine-induced hepatic carcinomas defined with surface-reactive monoclonal antibodies. AB - To gain further insight into the differentiation of oval cells and their role in carcinogenesis, we have generated cell surface reactive monoclonal antibodies (MAbs) by a Balb/c nude mouse (nu/nu) immunization protocol. Three MAbs designated OC.4, OC.5, and OC.10 were generated from a mouse immunized with CDE6, an oval cell line established from oval cells induced by feeding a choline deficient diet containing 0.1% ethionine (CDE). These MAbs demonstrated stage specific expression in fetal liver and displayed strong reactivity with oval and bile duct epithelial cells. In general, oval cells displayed a more mature phenotype than fetal ductal cells, suggesting the existence in adult liver of more primitive ductal progenitors. A fourth MAb recognized a cytoplasmic antigen (OC.6) expressed by mucus-secreting hepatic ducts induced by CDE diet. Immunocytochemical analysis indicated that OC.4, OC.5, and OC.10 were also expressed on CDE-induced, OV6+ hepatocellular carcinomas (HCC) but not on OV6+ HCC induced by the Solt/Farber protocol. In most cases, CDE-induced, OV6+ HCC expressed early ductal developmental markers such as OC.10 but lacked those expressed at later stages (OC.5, OC.4). These new MAb will be useful for characterizing HCC subpopulations with oval cell characteristics and for isolating biliary cells at antigenically defined stages during differentiation. PMID- 10816385 TI - Retinoic acid affects the EGF-R signaling pathway during differentiation induction of human endometrial adenocarcinoma cells. AB - We have shown that moderately differentiated endometrial adenocarcinoma (RL95-2) cells differentiate in response to retinoic acid treatment, illustrated by their reorganization of actin filaments and cell enlargement (Carter et al., Anticancer Res. 16, 17-24, 1996). Tyrphostin, an inhibitor of epidermal growth factor receptor (EGF-R)-associated protein tyrosine kinases, caused a dramatic reorganization of actin filaments in RL95-2 cells, similar to retinoic-acid treated cells (Carter and Bellido, J. Cell. Physiol. 178, 320-332, 1999). We evaluated the possibility that the differentiating effects of retinoids are due to retinoic-acid-induced decreases in phosphorylation of EGF-R and changes in downstream effector proteins. Retinoic acid caused a decrease in tyrosine phosphorylation of EGF-R. Retinoic acid treatment induced a dramatic actin filament reorganization and cell enlargement. Treatment with EGF reversed this effect, because cells treated with retinoic acid followed by EGF only possessed disrupted actin aggregates and appeared small, thus resembling medium controls. Retinoic acid induced a relocalization and decrease in the amount of Shc protein, another actin-binding protein which is an adaptor protein for EGF-R signaling. In addition, retinoic acid induced a relocalization of gelsolin from the plasma membrane to the cytoplasm. Retinoic acid decreased cell detachment in detachment assays; one-half as many retinoic-acid-treated cells detached as in controls. These results are consistent with the idea that retinoic acid induces differentiation of RL95-2 cells by interfering with the EGF-R signaling pathway. PMID- 10816386 TI - Expression of protein tyrosine phosphatases and its significance in esophageal cancer. AB - Expression of mRNA protein tyrosine phosphatases (PTPs) was surveyed in an esophageal cancer cell line by RT-PCR using degenerate primers. The mRNAs for eight kinds of PTPs were expressed in the cell line. We examined mRNA expression of these PTPs in 12 cases of esophageal cancer by Northern analysis. Significant signals were obtained for three kinds of PTPs, PTP1B, PTPH1, and PTPD1. The magnitude of expression of each PTP was measured as the ratio of the signal intensity of each PTP to that of a control gene (NADPH), and the ratio was then compared to normal mucosa around the cancer lesion. Among the three kinds of PTPs, the expression of PTP1B mRNA was significantly depressed in cancer lesions compared with that in the surrounding normal mucosa. In contrast, the expression of PTPH1 mRNA was significantly increased in cancer lesions compared with that in normal mucosa. PTPD1 did not show any significant trend in comparisons of cancer and surrounding normal mucosa. The results suggest that PTP1B and PTPH1 are engaged in opposing signaling pathways, the tumor-suppressive and tumor-promoting pathways, respectively, in esophageal carcinogenesis. PMID- 10816388 TI - Reply PMID- 10816387 TI - Re: Antibodies to squalene in Gulf War Syndrome. PMID- 10816390 TI - The ultrasonic barberpole: midgut volvulus and malrotation in a young adult PMID- 10816389 TI - Editorial note PMID- 10816391 TI - Air injection during FNA-ultrasound and mammographic appearances PMID- 10816392 TI - Giant myelolipoma of the adrenal gland: natural history PMID- 10816394 TI - Adrenaline in reactions to intravenous contrast medium PMID- 10816393 TI - Reply to dr troughton PMID- 10816396 TI - Publisher's announcement PMID- 10816395 TI - UNENHANCED HELICAL CT FOR RENAL COLIC - IS THE RADIATION DOSE JUSTIFIABLE? PMID- 10816397 TI - Pulmonary complications following lung transplantation. AB - Lung transplantation has become an accepted procedure for the treatment of end stage lung disease, being successful in improving the quality and length of life of many patients. The recognition and early treatment of complications is important for long-term survival of lung transplant recipients. The radiologist frequently plays a central role in investigation. The radiological appearance of pulmonary complications arising in the immediate, early (within 2 months) and late (after 2 months) stages post-transplantation are reviewed. PMID- 10816398 TI - The clinical and radiological features of Fanconi's anaemia. AB - Fanconi's anaemia is a severe refractory anaemia, associated with congenital malformations in approximately two-thirds of cases. Although these malformations may involve every organ system, suggestive dysmorphic features include growth retardation, radial ray deformities and urinary malformations. These malformations are not specific for Fanconi's anaemia, but should be recognized during pregnancy, or later in childhood, and suggest the possibility of inherited haematopoiesis disorders. PMID- 10816399 TI - Assessment of renal artery stenosis: comparison of captopril renography and gadolinium-enhanced breath-hold MR angiography. AB - AIM: To determine the accuracy of captopril renography (CR) and gadolinium enhanced breath-hold magnetic resonance (MR) angiography in the diagnosis of 50 99% renal artery stenosis (RAS). MATERIALS AND METHODS: Forty-three patients with possible RAS, of whom 53% had renal function impairment (creatinine >130 micromol/l), were included.(99m)Tc-mercaptoacetyl triglycine (MAG(3)) renography was performed after an oral dose of 25 mg captopril. Gadolinium-enhanced MR angiography was performed on a standard 1.5 Tesla system: TR 13.5, TE 3.5, flip angle 60 degrees, matrix 195 x 512. Intra-arterial digital subtraction angiography (DSA) was the standard of reference. RESULTS: Captropril renography accurately categorized 22 of 26 patients who had either uni- or bilateral RAS of 50-99%. The sensitivity and specificity of CR for the detection of 50-99% stenosis were 85 and 71%, respectively. With MR angiography one occluded artery was incorrectly diagnosed as a stenosis. Sensitivity and specificity were 100 and 94%, respectively. The difference between the accuracies of MR angiography and CR was statistically significant (P = 0.02). The accuracy of CR was lower in patients with renal impairment (70%) than in those with normal renal function (90%). CONCLUSION: MR angiography showed a high accuracy in diagnosing RAS of between 50 and 99%. CR was less accurate than MR angiography, especially in patients with renal function impairment. In patients with normal renal function, however, CR remains a useful diagnostic test. PMID- 10816400 TI - Symptomatic and asymptomatic accessory navicular bones: findings of Tc-99m MDP bone scintigraphy. AB - AIM: The accuracy of bone scintigraphy in diagnosing symptomatic accessory navicular bones has not been well studied. We conducted a retrospective study to explore the results and use of scintigraphy in symptomatic and asymptomatic accessory navicular bones. MATERIALS AND METHODS: Thirteen patients with a total of 13 symptomatic and 10 asymptomatic accessory navicular bones were included in the study. We used a scoring system to grade the scintigraphic abnormalities. The patients' symptoms and scintigraphic findings were recorded. RESULTS: Though focally increased radiopharmaceutical uptake was observed in all symptomatic accessory naviculars, half of the asymptomatic accessory navicular bones had the same manifestations. The scoring system was of no value in differentiating symptomatic from asymptomatic accessory navicular bones. CONCLUSION: Bone scintigraphy is a sensitive but not a specific tool for diagnosing a symptomatic accessory navicular. PMID- 10816401 TI - Prospective evaluation of ultrasound in distal ileal and colonic obstruction. AB - AIM: To evaluate prospectively the usefulness of ultrasound in determining the site and cause of distal small bowel and colonic obstruction. MATERIALS AND METHODS: Ultrasound findings in 60 consecutive patients with suspected distal ileal or colonic obstruction were correlated with final surgical and radiological diagnoses. The diagnostic value of ultrasound was compared with plain abdominal radiography (AXR) for the presence and level of obstruction. RESULTS: Forty-five patients were confirmed to be obstructed. Ultrasound correctly identified obstruction in 44/45 and the cause in 36/45 (80%). Overall sensitivity of US for obstruction was 98% and specificity 80%, compared with 79% and 53% respectively for the AXR. CONCLUSION: Ultrasound is useful in determining the presence and cause of distal ileal and colonic obstruction. PMID- 10816402 TI - Facial lymphadenopathy in nasopharyngeal carcinoma. AB - AIMS: This paper reports the findings of facial nodal metastasis in nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: The film records of 1916 patients with histologically confirmed NPC seen over a 5-year period were reviewed. RESULTS: Eight facial nodes were demonstrated in three (0.2%) patients. There were three buccinator, two malar, two infraorbital and one mandibular nodes. CONCLUSION: Facial nodal metastasis in NPC is unusual, but may be seen at presentation or during recurrence. PMID- 10816403 TI - Optimized enhancement in helical CT: experiences with a real-time bolus tracking system in 628 patients. AB - AIMS: Ultrafast detector technology enables bolus-triggered application of contrast media. In a prospective study we investigated the benefit of this new method with the intention of optimizing enhancement during examination of the chest and abdomen. MATERIALS AND METHODS: In total, we examined 548 patients under standardized conditions. All examinations were performed on a Somatom Plus 4 Power CT system (Siemens Corp., Forchheim, Germany) using the CARE-Bolus software. This produces repetitive low-dose test images (e.g. for the lung: 140 kV, 43 mA, TI 0.5 s) and measures the Hounsfield attenuation in a pre-selected region of interest. After exceeding a defined threshold, a diagnostic spiral CT examination was begun automatically. The data obtained from 321 abdominal CT and 179 lung CT examinations were correlated with different parameters such as age, weight and height of the patients and parameters of vascular access. In a group of 80 patients, the injection of contrast medium was stopped after reaching a pre defined threshold of an increase of 100 HU over the baseline. Then, we assessed the maximal enhancement of liver, pulmonal artery trunk and aortic arch. RESULTS: There was no correlation between bolus geometry and age, body surface or weight. In helical CT of the abdomen the threshold was reached after a mean trigger time of 27 s (range 13-67 s) and only 65 ml (range 41-105 ml) of contrast medium were administered. In helical CT of the lung the threshold was reached after 21 s (range 12-48 s) and the mean amount of administered contrast medium was 48 ml (range 38-71 ml). CONCLUSION: Bolus triggering allows optimized enhancement of the organs and reduces the dose of contrast material required compared with standard administration. PMID- 10816404 TI - Digital imaging improves upright stereotactic core biopsy of mammographic microcalcifications. AB - AIM: This comparative study was carried out to assess the effect of using digital images compared to conventional film-screen mammography on the accuracy of core biopsy of microcalcifications using upright stereotactic equipment. MATERIALS AND METHODS: The biopsy results from a consecutive series of 104 upright stereotactic 14-gauge core biopsies performed with conventional X-ray (Group A) were compared with 40 biopsies carried out using stereotaxis with digital imaging (Group B). In all cases specimen radiography was performed and analysed for the presence of calcifications. Pathological correlation was then carried out with needle and surgical histology. RESULTS: The use of digital add-on equipment increased the radiographic calcification retrieval rate from 55 to 85% (P < 0.005). The absolute sensitivity of core biopsy in pure ductal carcinoma in situ (DCIS) cases rose from 34 to 69% (P < 0.03), with the complete sensitivity increasing from 52 to 94% (P < 0.005). For DCIS with or without an invasive component the absolute sensitivity rose from 41 to 67% (P = 0.052), while the complete sensitivity was 59% before and 86% after the introduction of digital imaging (P < 0.04). CONCLUSION: Digital equipment improves the performance of upright stereotactic core biopsy of microcalcifications, giving a significantly increased success rate in accurately obtaining calcifications. This leads to an improvement in absolute and complete sensitivity of core biopsy when diagnosing DCIS. PMID- 10816405 TI - Choledochal cyst: comparison of MR and conventional cholangiography. AB - AIMS: To assess the diagnostic value of magnetic resonance (MR) cholangiography versus conventional cholangiography in patients with choledochal cyst and to determine whether MR cholangiography can be considered an alternative to conventional cholangiography. MATERIALS AND METHODS: Thirteen patients with choledochal cyst were examined by MR cholangiography and conventional cholangiograms. Magnetic resonance cholangiography employed T2-weighted axial and coronal fast spin-echo, single and multislab single-shot fast spin-echo sequences, including source images with maximum intensity projections. The diagnostic value of MR cholangiography and conventional cholangiograms was assessed and compared using the criteria of depiction of morphology, anomalous pancreaticobiliary duct union and demonstration of complications such as stones. A four-point diagnostic scale was applied to the delineation of the ductal anatomy with the Wilcoxon signed-ranks test and McNemar's test used for statistical analysis. RESULTS: The depiction of the choledochal cyst was significantly better with MR cholangiography than with conventional cholangiography (P = 0.03). The detection rate of an anomalous pancreaticobiliary duct union was not significantly different with either method (P = 0.641), nor was the detection rate of bile duct stones (P = 0.375). CONCLUSION: Magnetic resonance cholangiography provides data equivalent to or superior to those from conventional cholangiography in evaluating choledochal cyst. Magnetic resonance cholangiography is recommended as a non-invasive examination of choice for the evaluation of choledochal cyst. PMID- 10816406 TI - Low grade gastric MALT Lymphoma: radiographic findings. AB - AIMS: Gastric MALT (mucosa-associated lymphoid tissue) lymphoma is now recognized as a distinct entity within extranodal non-Hodgkin's lymphoma. The purpose of this study was to describe the radiographic findings in low grade gastric MALT lymphoma. MATERIALS AND METHODS: We retrospectively reviewed the radiographic findings in 22 cases of low-grade gastric MALT lymphoma. The study group consisted of 15 men and seven women (median age 68 years, range 41-91 years). Lesions were designated as infiltrative or polypoid by consensus of two radiologists. Polypoid lesions were categorized by number and size. Anatomical site within the stomach and presence of transpyloric or oesophagogastric extension was determined for each case. The presence of abdominal lymphadenopathy was categorized as regional or distant. The presence of Helicobacter pylori was determined from endoscopic and surgical biopsies. RESULTS: Computed tomography (CT) revealed abnormalities of the stomach in 19 cases of the 21 in which it was performed. There were 14 infiltrative lesions and five polypoid lesions. Of the 14 infiltrative lesions, the mean gastric wall thickness was 2.2 cm (range 0.8 6.0 cm). There were three single and two multiple polypoid lesions (mean size 2.2 cm, range 1. 5-2.7 cm). Transpyloric extension was observed in two cases and oesophagogastric extension in one. Abdominal lymphadenopathy was observed in 10 of 21 patients. Helicobacter pylori was found in 19 of 22 cases (86%). CONCLUSION: Low grade B cell gastric MALT lymphomas present with an infiltrative form on CT in about three-quarters of cases and a polypoid pattern in the remainder. Abdominal lymphadenopathy is seen in approximately one-half of cases. There is a high association with Helicobacter pylori. PMID- 10816407 TI - Ultrasound guided core biopsy of suspicious mammographic calcifications using high frequency and power Doppler ultrasound. AB - AIM: The pre-operative diagnosis of suspicious mammographic microcalcifications usually requires stereotactic needle biopsy. The aim of this study was to evaluate if high frequency 13 MHz ultrasound (HFUS) and power Doppler (PD) can aid visualization and biopsy of microcalcifications. MATERIALS AND METHODS: Forty four consecutive patients presenting with microcalcifications without associated mammographic or palpable masses were examined with HFUS and PD. Ultrasound-guided core biopsy (USCB) was performed where possible. Stereotactic biopsy was carried out when US-guided biopsy was unsuccessful. Surgery was performed if a diagnosis of malignancy was made on core biopsy or if the repeat core biopsy was non diagnostic. RESULTS: Forty-one patients (93%) had ultrasound abnormalities corresponding to mammographic calcification. USCB was performed on 37 patients. In 29/37, USCB obtained a definitive result (78.4%). USCB was non-diagnostic in 4/9 benign (44.4%) and 4/28 (14.3%) malignant lesions biopsied. The complete and absolute sensitivities for malignancy using USCB were 85.7% (24/28) and 81% (23/28), respectively. USCB correctly identified invasive disease in 12/23 (52.2%) cases. There was no significant difference in the presence of abnormal flow on PD between benign and malignant lesions. However, abnormal PD vascularity was present in 43.5% of invasive cancer and was useful in directing successful biopsy in eight cases. CONCLUSION: The combination of high frequency US with PD is useful in the detection and guidance of successful needle biopsy of microcalcifications particularly where there is an invasive focus within larger areas of DCIS. PMID- 10816408 TI - The effect of timing of intravenous muscle relaxant on the quality of double contrast barium enema. AB - AIM: To determine whether the timing of buscopan administration during double contrast barium enema examination (DCBE) affects diagnostic quality. MATERIALS AND METHODS: In a prospective setting, 100 consecutive adult out-patients referred for DCBE received 20 mg buscopan (hyoscine-N-butylbromide) intravenously, either before infusion of barium suspension (Group A) or after barium infusion and gas insufflation (Group B). A subjective assessment of ease of contrast medium infusion was made at the time of examination and the films subsequently analysed by two radiologists unaware of the mode of relaxant administration, who noted the quality of mucosal coating and made subjective and objective measurements of segmental distension. RESULTS: There was no significant difference in screening times, infusion difficulty or colonic contrast medium coating between the two groups. Subjective assessment of distension of the caecum, ascending colon, transverse colon and rectum were not significantly different. Patients receiving intravenous relaxant after barium and gas infusion had less subjective descending (P = 0. 05) and sigmoid (P = 0.04) colon distension, but there was no significant difference with respect to maximal bowel diameter in any of the segments measured. CONCLUSION: The timing of intravenous administration during DCBE is likely to have no significant effect on the diagnostic quality of the study. PMID- 10816409 TI - Hodgkin's lymphoma of the urinary bladder. PMID- 10816410 TI - Metastatic liposarcoma of the brain. PMID- 10816411 TI - Conservative surgical removal of submandibular duct calculus following interventional sialography. PMID- 10816412 TI - Encyclopaedia of medical imaging, vol. II PMID- 10816413 TI - Living with radiation PMID- 10816414 TI - Differential diagnosis in head and neck imaging PMID- 10816415 TI - Breast ultrasound PMID- 10816417 TI - Inhibition of glutaminase expression by antisense mRNA decreases growth and tumourigenicity of tumour cells. AB - Phosphate-activated glutaminase has a critical role in tumours and rapidly dividing cells and its activity is correlated with malignancy. Ehrlich ascites tumour cells transfected with the pcDNA3 vector containing an antisense segment (0.28 kb) of rat kidney glutaminase showed impairment in the growth rate and plating efficiency, as well as a shortage in the glutaminase protein and activity. The C-terminal segment used is well conserved in all glutaminase sequences known. The transfected cells, named 0.28AS-2, displayed remarkable changes in their morphology compared with the parental cell line. The 0.28AS-2 cells also lost their tumourigenic capacity in vivo. Control mice developed an ascitic tumour, with a lifespan of 16+/-1 days, when inoculated with 10(7) cells/mouse; on the contrary, animals inoculated with transfected cells up to 2.5 times the cell numbers of control mice did not develop tumours and behaved as healthy animals. The ability to revert the transformed phenotype of antisense transfected cells confirms the relevance of glutaminase in the transformation process and could provide new ways for the study of gene therapy. PMID- 10816418 TI - Saccharomyces cerevisiae Yak1p protein kinase autophosphorylates on tyrosine residues and phosphorylates myelin basic protein on a C-terminal serine residue. AB - The serine/threonine protein kinase, Yak1p, functions as a negative regulator of the cell cycle in Saccharomyces cerevisiae, acting downstream of the cAMP dependent protein kinase. In the present work we report that overexpression of haemagglutinin-tagged full-lengthYak1p and an N-terminally truncated form (residues 148-807) lead to growth arrest in PKA compromised yak1 null yeast cells. Both forms of recombinant Yak1p kinase were catalytically active and preferred myelin basic protein (MBP) as a substrate over several other proteins. Phosphopeptide analysis of bovine MBP by tandem MS revealed two major Yak1p phosphorylation sites, Thr-97 and Ser-164. Peptides containing each site were obtained and tested as Yak1p substrates. Both forms of Yak1p phosphorylated a peptide containing the Ser-164 residue with far more efficient kinetics than MBP. The maximal velocity (V(max)) values of the full-length Yak1p reaction were 110+/ 21 (Ser-164) and 8.7+/-1.7 (MBP), and those of N-terminally truncated Yak1p were 560.7+/-74.8 (Ser-164) and 34. 4+/-2.2 (MBP) pmol/min per mg of protein. Although neither form of Yak1p was able to phosphorylate two generic protein tyrosine kinase substrates, both were phosphorylated on tyrosine residues in vivo and underwent tyrosine autophosphorylation when reacted with ATP in vitro. Tandem MS showed that Tyr-530 was phosphorylated both in vivo and in vitro after reaction with ATP. Pre-treatment with protein tyrosine phosphatase 1B removed all of Yak1p phosphotyrosine content and drastically reduced Yak1p activity against exogenous substrates, suggesting that the phosphotyrosine content of the enzyme is essential for its catalytic activity. Although the N-terminally truncated Yak1p was expressed at a lower level than the full-length protein, its catalytic activity and phosphotyrosine content were significantly higher than those of the full-length enzyme. Taken together, our results suggest that Yak1p is a dual specificity protein kinase which autophosphorylates on Tyr-530 and phosphorylates exogenous substrates on Ser/Thr residues. PMID- 10816416 TI - Rho GTPases and their effector proteins. AB - Rho GTPases are molecular switches that regulate many essential cellular processes, including actin dynamics, gene transcription, cell-cycle progression and cell adhesion. About 30 potential effector proteins have been identified that interact with members of the Rho family, but it is still unclear which of these are responsible for the diverse biological effects of Rho GTPases. This review will discuss how Rho GTPases physically interact with, and regulate the activity of, multiple effector proteins and how specific effector proteins contribute to cellular responses. To date most progress has been made in the cytoskeleton field, and several biochemical links have now been established between GTPases and the assembly of filamentous actin. The main focus of this review will be Rho, Rac and Cdc42, the three best characterized mammalian Rho GTPases, though the genetic analysis of Rho GTPases in lower eukaryotes is making increasingly important contributions to this field. PMID- 10816419 TI - Tyrosine phosphorylation of the vascular endothelial-growth-factor receptor-2 (VEGFR-2) is modulated by Rho proteins. AB - The effects of Rho-specific modifying toxins on the tyrosine phosphorylation of endothelial cell proteins were investigated. Incubation of the cells with the Rho activating toxin cytotoxic necrotizing factor 1 (CNF1) induced a marked increase in the tyrosine phosphorylation of a number of signalling intermediates of the vascular endothelial growth factor (VEGF)-mediated cascade, including focal adhesion kinase, paxillin, phospholipase Cgamma1 and a Shc-associated protein of 195 kDa. Both CNF1- and VEGF-dependent tyrosine phosphorylation of these proteins were significantly reduced by prior incubation with C3 transferase, a known inhibitor of RhoA function, suggesting a Rho-dependent mechanism. The stimulation of endothelial cells with CNF1 resulted in a marked increase in the tyrosine phosphorylation of the VEGF receptor (VEGFR)-2, which was correlated with a stimulation of its kinase activity and with its association with downstream tyrosine phosphorylated proteins. The stimulatory effect of CNF1 was specific for VEGFR-2 since the phosphotyrosine content of VEGFR-1 was not affected by the toxin. Transient overexpression of a dominant-active RhoA mutant also induced an increase in the tyrosine phosphorylation of the VEGFR-2, whereas overexpression of a dominant-inactive form of the protein was without effect. Taken together, these results indicate that Rho proteins may play an important role in angiogenesis by modulating the tyrosine phosphorylation levels of VEGFR-2. PMID- 10816420 TI - Human Sug1/p45 is involved in the proteasome-dependent degradation of Sp1. AB - The transcription factor Sp1 was previously shown to undergo proteasome-dependent degradation when cells were glucose-starved and stimulated with the adenylate cyclase inducer, forskolin. However, the control of the Sp1 degradation process is largely unknown. Using in vitro and in vivo interaction studies, we show in the present study that Sp1 interacts with human Sug1 [hSug1, also known as p45 or thyroid-hormone-receptor interacting protein ('TRIP1')], an ATPase subunit of the 26 S proteasome and a putative transcriptional modulator. This interaction with Sp1 occurs through the C-terminus of hSug1, the region that contains the conserved ATPase domain in this protein. Both in vitro studies, in reconstituted degradation assays, and in vivo experiments, in which hSug1 is overexpressed in normal rat kidney cells, show that full-length hSug1 is able to stimulate the proteasome-dependent degradation of Sp1. However, hSug1 truncations that lack either the N- or C-terminal domain of hSug1 act as dominant negatives, inhibiting Sp1 degradation in vitro. Also, an ATPase mutant of hSug1, while still able to bind Sp1, acts as a dominant negative, blocking Sp1 degradation both in vitro and in vivo. These results demonstrate that hSug1 is involved in the degradation of Sp1 and that ATP hydrolysis by hSug1 is necessary for this process. Our findings indicate that hSug1 is an exchangeable proteasomal component that plays a critical regulatory role in the proteasome-dependent degradation of Sp1. However, hSug1 is not the factor limiting Sp1 degradation in the cells treated with glucosamine. This and other considerations suggest that hSug1 co-operation with other molecules is necessary to target Sp1 for proteasome degradation. PMID- 10816421 TI - CLC-Nt1, a putative chloride channel protein of tobacco, co-localizes with mitochondrial membrane markers. AB - The voltage-dependent chloride channel (CLC) family of membrane proteins has cognates in animals, yeast, bacteria and plants, and chloride-channel activity has been assigned to most of the animal homologues. Lack of evidence of CLC functions in plants prompted us to characterize the cellular localization of the tobacco CLC-Nt1 protein. Specific polyclonal antibodies were raised against an N terminal polypeptide of CLC-Nt1. These antibodies were used to probe membrane proteins prepared by various cell-fractionation methods. These included aqueous two-phase partitioning (for plasma membranes), free-flow electrophoresis (for vacuolar and plasma membranes), intact vacuole isolation, Percoll-gradient centrifugation (for plastids and mitochondria) and stepped, linear, sucrose density-gradient centrifugation (for mitochondria). Each purified membrane fraction was characterized with specific marker enzyme activities or antibodies. Our studies ruled out the possibility that the major cell localization of CLC-Nt1 was the vacuolar or plasma membranes, the endoplasmic reticulum, the Golgi apparatus or the plastids. In contrast, we showed that the tobacco CLC-Nt1 specifically co-localized with the markers of the mitochondrial inner membrane, cytochrome c oxidase and NAD9 protein. CLC-Nt1 may correspond to the inner membrane anion channel ('IMAC') described previously in animal and plant mitochondria. PMID- 10816422 TI - Chicken Y-box proteins chk-YB-1b and chk-YB-2 repress translation by sequence specific interaction with single-stranded RNA. AB - Y-Box proteins comprise a large family of multifunctional proteins with a wide spectrum of activities in both transcription and translational regulation of gene expression. Earlier, we have reported on the involvement of chk-YB-2 in transcriptional regulation of Rous sarcoma virus long terminal repeats and the involvement of chk-YB-1b in transcriptional regulation of alpha1(I) collagen genes. Here, we have investigated the potential role of chk-YB-2 and chk-YB-1b in RNA metabolism. We report that chk-YB-2 and chk-YB-1b are localized predominantly in the cytoplasm and that they both can bind single-stranded RNA in a sequence specific and reversible manner. Well-conserved cold-shock domain, N-terminal proline-rich domain and the alternating clusters of acidic and basic amino acids located in the C-terminal ends of these two proteins were all found to be necessary for their RNA-binding ability. Further, we demonstrate that these two proteins inhibit translation in vitro and that binding to RNA is required for this inhibition. The significance of these results is discussed. PMID- 10816423 TI - Cholesterol decreases secretion of the secreted form of amyloid precursor protein by interfering with glycosylation in the protein secretory pathway. AB - Cerebral deposits of beta-amyloid (betaA) are a major feature in Alzheimer's disease. betaA is derived from amyloid precursor protein (APP). APP is subject to N- and O-glycosylation and undergoes a series of proteolytic cleavages that lead to the release of betaA or of a non-amyloidogenic secreted form of APP (APPs). We used primary neuronal and glial cultures to investigate how cholesterol affects the production and secretion of APPs. Exposure to cholesterol for 2 h did not change the neuronal release of APPs; after 6 h APPs release was slightly lower, whereas 24 h of exposure decreased APPs in the medium by approx. 60%. The time courses were similar in astrocytes and microglia preparations. To verify whether the effect of cholesterol was a consequence of membrane rigidification we tested the activity of ganglioside GM1 and prion protein fragment PrP 106-126, which affect membrane fluidity similarly to cholesterol, on APPs secretion. Neither altered the production of APPs. APP mRNA and the total amount of APP in the cells were slightly decreased by cholesterol after 2 and 24 h respectively. Immunoblot analysis of APP associated with neuronal cells and astrocytes indicated that cholesterol progressively decreased the glycosylated forms of the protein; a similar tendency was noted in cells treated with brefeldin A and monensin, two substances that interfere with protein glycosylation. The cell-surface biotinylation method showed that in cholesterol-treated cells APP reached the plasma membrane. Our results indicate that cholesterol decreases the secretion of APPs by interfering with APP maturation and inhibiting glycosylation of the protein; although APP is inserted in the membrane it is not cleaved by alpha secretase. PMID- 10816424 TI - Oxalomalate, a competitive inhibitor of aconitase, modulates the RNA-binding activity of iron-regulatory proteins. AB - We investigated the effect of oxalomalate (OMA, alpha-hydroxy-beta-oxalosuccinic acid), a competitive inhibitor of aconitase, on the RNA-binding activity of the iron-regulatory proteins (IRP1 and IRP2) that control the post-transcriptional expression of various proteins involved in iron metabolism. The RNA-binding activity of IRP was evaluated by electrophoretic mobility-shift assay of cell lysates from 3T3-L1 mouse fibroblasts, SH-SY5Y human cells and mouse livers incubated in vitro with OMA, with and without 2-mercaptoethanol (2-ME). Analogous experiments were performed in vivo by prolonged incubation (72 h) of 3T3-L1 cells with OMA, and by injecting young mice with equimolar concentrations of oxaloacetate and glyoxylate, which are the precursors of OMA synthesis. OMA remarkably decreased the binding activity of IRP1 and, when present, of IRP2, in all samples analysed. In addition, the recovery of IRP1 by 2-ME in the presence of OMA was constantly lower versus control values. These findings suggest that the severe decrease in IRP1 RNA-binding activity depends on: (i) linking of OMA to the active site of aconitase, which prevents the switch to IRP1 and explains resistance to the reducing agents, and (ii) possible interaction of OMA with some functional amino acid residues in IRP that are responsible for binding to the specific mRNA sequences involved in the regulation of iron metabolism. PMID- 10816425 TI - Kinetic study of the inactivation of ascorbate peroxidase by hydrogen peroxide. AB - The activity of ascorbate peroxidase (APX) has been studied with H(2)O(2) and various reducing substrates. The activity decreased in the order pyrogallol>ascorbate>guaiacol>2, 2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS). The inactivation of APX with H(2)O(2) as the sole substrate was studied. The number of H(2)O(2) molecules required for maximal inactivation of the enzyme was determined as approx. 2.5. Enzymic activity of approx. 20% of the original remained at the end of the inactivation process (i.e. approx. 20% resistance) when ascorbate or ABTS was used as the substrate in activity assays. With pyrogallol or guaiacol no resistance was seen. Inactivation by H(2)O(2) followed over time with ascorbate or pyrogallol assays exhibited single exponential decreases in enzymic activity. Hyperbolic saturation kinetics were observed in both assay systems; a similar dissociation constant (0.8 microM) for H(2)O(2) was obtained in each case. However, the maximum rate constant (lambda(max)) obtained from the plots differed depending on the assay substrate. The presence of reducing substrate in addition to H(2)O(2) partly or completely protected the enzyme from inactivation, depending on how many molar equivalents of reducing substrate were added. An oxygen electrode system has been used to confirm that APX does not exhibit a catalase-like oxygen-releasing reaction. A kinetic model was developed to interpret the experimental results; both the results and the model are compared and contrasted with previously obtained results for horseradish peroxidase C. The kinetic model has led us to the conclusion that the inactivation of APX by H(2)O(2) represents an unusual situation in which no enzyme turnover occurs but there is a partition of the enzyme between two forms, one inactive and the other with activity towards reducing substrates such as ascorbate and ABTS only. The partition ratio is less than 1. PMID- 10816426 TI - Positional- and stereo-selectivity of fatty acid oxygenation catalysed by mouse (12S)-lipoxygenase isoenzymes. AB - A quantitative stereochemical analysis of the products generated by recombinant mouse (12S)-lipoxygenase isoenzymes was performed with arachidonic acid and linoleic acid as substrates. The leucocyte-type (12S)-lipoxygenase generated, in addition to 12-hydroxyeicosatetraenoic acid (12-HETE) as the main product, 15- and 8-HETE from arachidonic acid and 13- and 9-hydroxyoctadecadienoic acid (13- and 9-HODE) from linoleic acid. The platelet-type enzyme oxygenated arachidonic acid to 12- and 8-HETE and linoleic acid to 13- and 9-HODE, whereas the epidermis type (12S)-lipoxygenase reaction was essentially mono-specific with arachidonic acid but oxygenated linoleic acid to both 13- and 9-HODE. 12-HETE and 13-HODE were almost exclusively the S enantiomers. 8-HETE was the R enantiomer as a side product of the platelet-type (12S)-lipoxygenase reaction but the S enantiomer as a side-product of the leucocyte-type reaction. 9-HODE was generated as the R enantiomer by the platelet-type and the epidermis-type isoenzymes and as the S enantiomer by the leucocyte-type (12S)-lipoxygenase. On the basis of published models of lipoxygenase-substrate interaction, the stereochemistry of the products generated by the platelet- and epidermis-type (12S)-lipoxygenases is in agreement with a fixed 'tail-to-head' orientation of the substrate fatty acid in the binding pocket of these enzymes, whereas that of the reaction products of the leucocyte-type (12S)-lipoxygenase can be explained only when the inverse orientation of the substrate or a rotational isomerism along the longitudinal axis of the substrate is allowed. Both the product spectra generated and the sensitivity towards the 12-lipoxygenase selective inhibitors N-benzyl-N-hydroxy-4 phenylpentanamide and cinnamyl-3,4-dihydroxy-alpha-cyanocinnamate indicated the platelet-type and the epidermis-type isoenzymes to be biochemically more related to each other than to the leucocyte-type (12S)-lipoxygenase. PMID- 10816427 TI - Expression of an active form of recombinant Ty1 reverse transcriptase in Escherichia coli: a fusion protein containing the C-terminal region of the Ty1 integrase linked to the reverse transcriptase-RNase H domain exhibits polymerase and RNase H activities. AB - Replication of the Saccharomyces cerevisiae Ty1 retrotransposon requires a reverse transcriptase capable of synthesizing Ty1 DNA. The first description of an active form of a recombinant Ty1 enzyme with polymerase and RNase H activities is reported here. The Ty1 enzyme was expressed as a hexahistidine-tagged fusion protein in Escherichia coli to facilitate purification of the recombinant protein by metal-chelate chromatography. Catalytic activity of the recombinant protein was detected only when amino acid residues encoded by the integrase gene were added to the N-terminus of the reverse transcriptase-RNase H domain. This suggests that the integrase domain could play a role in proper folding of reverse transcriptase. Several biochemical properties of the Ty1 enzyme were analysed, including the effect of MgCl(2), NaCl, temperature and of the chain terminator dideoxy GTP on its polymerase activity. RNase H activity was examined by monitoring the cleavage of a RNA-DNA template-primer. Our results suggest that the distance between the RNase H and polymerase active sites corresponds to the length of a 14-nucleotide RNA-DNA heteroduplex. The recombinant protein produced in E. coli should be useful for further biochemical and structural analyses and for a better understanding of the role of integrase in the activation of reverse transcriptase. PMID- 10816428 TI - Cytochrome c release from isolated rat liver mitochondria can occur independently of outer-membrane rupture: possible role of contact sites. AB - Percoll-purified rat liver mitochondria were shown to contain BAX dimer and rapidly (<2 min) release 5-10% of their cytochrome c when incubated in a standard KCl incubation medium under energized conditions. This release was not accompanied by release of adenylate kinase (AK), another intermembrane protein, and was not inhibited by Mg(2+), dATP, inhibitors of the permeability transition or ligands of the peripheral benzodiazepine receptor. However, release was greatly reduced by the presence of 5% (w/v) dextran (40 kDa), which caused a decrease in the light scattering (A(520)) of mitochondrial suspensions. Dextran also inhibited both mitochondrial oxidation of exogenous ferrocytochrome c in the presence of rotenone and antimycin, and respiratory-chain-driven reduction of exogenous ferricytochrome c. Hypo-osmotic medium or digitonin treatment of mitochondria caused a large additional release of both cytochrome c and AK that was not blocked by dextran. Polyaspartate, which stabilizes the low conductance state of the voltage-dependent anion channel (VDAC), increased cytochrome c release. VDAC and BAX are both found at the contact sites between the inner and outer membranes and dextran is known to stabilize these contact sites in isolated mitochondria. Thus our data suggest that regulation of a specific permeability pathway for cytochrome c may be mediated by changes in protein-protein interactions within contact sites. The adenine nucleotide translocase is known to bind to VDAC and thus provides an additional link between the specific cytochrome c release pathway and the permeability transition. PMID- 10816429 TI - Phosphatidylinositol 3-kinase, protein kinase B and ribosomal S6 kinases in the stimulation of thyroid epithelial cell proliferation by cAMP and growth factors in the presence of insulin. AB - The proliferation of most normal cells depends on the co-operation of several growth factors and hormones, each with a specific role, but the key events involved in the action of each necessary stimulant remain largely uncharacterized. In the present study, the pathways involved in the mechanism(s) of co-operation have been investigated in primary cultures of dog thyroid epithelial cells. In this physiologically relevant system, thyroid stimulating hormone (TSH) acting through cAMP, epidermal growth factor (EGF) and phorbol esters (such as PMA) induce DNA synthesis. Their effect requires stimulation of the insulin-like growth factor-1 (IGF-1) receptor by either IGF-1 or insulin, which are not themselves mitogenic agents. In contrast, hepatocyte growth factor (HGF) is itself fully mitogenic. The results of the study demonstrate that cAMP, EGF, HGF and PMA stimulate p70 ribosomal S6 kinase (p70 S6 kinase). However, insulin/IGF-1 also stimulate p70 S6 kinase. Thus stimulation of p70 S6 kinase might be necessary, but is certainly not sufficient, for the induction of DNA synthesis and is not specific for any stimulated pathway. In contrast, phosphatidylinositol 3-kinase (PI 3-kinase) and protein kinase B (PKB) activation by insulin and HGF is strong and sustained, whereas it is weak and transient with EGF and absent in the presence of TSH or PMA. These findings suggest that: (i) stimulation of PI 3-kinases and/or PKB is not involved in the cAMP-dependent pathways leading to thyrocyte proliferation, or in the action of PMA, (ii) the stimulation of the PI 3-kinase/PKB pathway may account for the permissive action of insulin/IGF-1 in the proliferation of these cells, and (iii) the stimulation of this pathway by HGF may explain why this agent does not require insulin or IGF 1 for its mitogenic action. PMID- 10816430 TI - Lipidation of apolipoprotein E influences its isoform-specific interaction with Alzheimer's amyloid beta peptides. AB - The inheritance of the apolipoprotein E (apoE) epsilon4 allele is a prevailing risk factor for sporadic and familial Alzheimer's disease (AD). ApoE isoforms bind directly to Alzheimer's amyloid beta (Abeta) peptides both in vitro and in vivo. Recent studies suggest that association of apoE with lipids may modulate its interaction with Abeta. We examined the binding of lipid-associated and delipidated apoE3 and apoE4 isoforms to Abeta utilizing a solid-phase binding assay and estimated the dissociation constants for the interaction of various apoE and Abeta species. Using native apoE isoforms from stably transfected RAW 264 and human embryonic kidney 293 cells, apoE3 had greater affinity than apoE4 for both Abeta1-40 and Abeta1-42. Delipidation of apoE decreased its affinity for Abeta peptides by 5-10-fold and abolished the isoform-specificity. Conversely, incorporation of apoE isoforms produced by baculovirus-infected Sf9 cells into reconstituted human high-density-lipoprotein lipoparticles restored the affinity values for Abeta peptides and resulted in preferential binding of apoE3. The data demonstrate that native lipid-associated apoE3 binds to Abeta peptides with 2-3 fold higher affinity than lipid-associated apoE4. Since the isoforms' binding efficiency correlate inversely with the risk of developing late-onset AD, the results suggest a possible involvement of apoE3 in the clearance or routing out of Abeta from the central nervous system as one of the mechanisms underlying the pathology of the disease. PMID- 10816431 TI - Interaction of Bacillus subtilis CsaA with SecA and precursor proteins. AB - CsaA from the Gram-positive bacterium Bacillus subtilis has been identified previously as a suppressor of the growth and protein-export defect of Escherichia coli secA(Ts) mutants. CsaA has chaperone-like activities in vivo and in vitro. To examine the role of CsaA in protein export in B. subtilis, expression of the csaA gene was repressed. While export of most proteins remained unaffected, export of at least two proteins was significantly reduced upon CsaA depletion. CsaA co-immunoprecipitates and co-purifies with the SecA proteins of E. coli and B. subtilis, and binds the B. subtilis preprotein prePhoB. Purified CsaA stimulates the translocation of prePhoB into E. coli membrane vesicles bearing the B. subtilis translocase, whereas it interferes with the SecB-mediated translocation of proOmpA into membrane vesicles of E. coli. The specific interaction with the SecA translocation ATPase and preproteins suggests that CsaA acts as a chaperone that promotes the export of a subset of preproteins in B. subtilis. PMID- 10816432 TI - Suppression of temperature-sensitivity of a dnaA46 mutant by excessive DNA supercoiling. AB - We report here that the high-temperature sensitivity of a dnaA46 mutant was suppressed by addition of high concentrations of NaCl into the culture medium. This suppression was also observed with other high-temperature-sensitive dnaA mutants, except dnaA167 and dnaA508 mutants, which have mutations in the N terminal region of DnaA protein. Since high concentrations of NaCl in the medium increased negative DNA supercoiling in a dnaA46 mutant, we hypothesized that the increase in DNA supercoiling is involved in the suppression of the temperature sensitivity of the dnaA46 mutant by high concentrations of NaCl. This hypothesis was supported by in vitro and in vivo results as follows. A low DNA replication activity of purified DnaA46 protein at high temperatures was increased in line with an increase in DNA supercoiling of template DNA. The dnaA46 mutant showed higher sensitivity to nalidixic acid, a DNA-relaxing drug, than did the wild-type cells under the conditions of high temperatures and high concentrations of NaCl. PMID- 10816433 TI - The m3 muscarinic acetylcholine receptor is coupled to mitogen-activated protein kinase via protein kinase C and epidermal growth factor receptor kinase. AB - The acetylcholine analogue carbachol rapidly activated mitogen-activated protein kinase (MAPK), and caused tyrosine phosphorylation of the adapter protein p52 Shc and the epidermalgrowth factor (EGF) receptor, in human embryonic kidney cells stably expressing m3 muscarinic receptors. The protein kinase C (PKC) inhibitor GF109203X caused a significant partial inhibition of m3 receptor-mediated activation of MAPK. The PKC-independent MAPK activity elicited by carbachol in the presence of GF109203X was reproducibly abolished by AG1478, an inhibitor of EGF-receptor tyrosine kinase activity, and by the Src tyrosine kinase inhibitor PP1. In a subset of these experiments, GF109203X concomitantly increased carbachol-induced tyrosine phosphorylation of p52 Shc and the EGF receptor. In co stimulation experiments, carbachol and EGF activated MAPK in a non-additive fashion; moreover, EGF-induced association of Shc with the phosphorylated EGF receptor was inhibited by carbachol. This effect of carbachol was blocked by GF109203X. The results indicate that MAPK activation by m3 receptor stimulation is regulated by two pathways; one dependent on PKC, and the other mediated via the EGF receptor and Src. Moreover, the EGF-receptor-dependent pathway may be subject to negative-feedback regulation via m3 receptor-coupled activation of PKC. PMID- 10816434 TI - Purification from rat liver of a novel constitutively expressed member of the aldo-keto reductase 7 family that is widely distributed in extrahepatic tissues. AB - Antiserum raised against human aflatoxin B(1) aldehyde reductase 1 (hAFAR1) has been used to identify a previously unrecognized rat aldo-keto reductase (AKR). This novel enzyme is designated rat aflatoxin B(1) aldehyde reductase 2 (rAFAR2) and it characteristically migrates faster during SDS/PAGE than does the archetypal ethoxyquin-inducible rAFAR protein (now called rAFAR1). Significantly, rAFAR2 is essentially unreactive with polyclonal antibodies raised against rAFAR1. Besides its distinct electrophoretic and immunochemical properties, rAFAR2 appears to be regulated differently from rAFAR1 as it is expressed in most rat tissues and does not appear to be induced by ethoxyquin. Multiple forms of rAFAR2 have been identified. Anion-exchange chromatography on Q-Sepharose, followed by adsorption chromatography on columns of Matrex Orange A and Cibacron Blue, have been employed to purify rAFAR2 from rat liver cytosol. The Q-Sepharose chromatography step resulted in the resolution of rAFAR2 into three peaks of AKR activity, two of which were purified and shown to be capable of catalysing the reduction of 2-carboxybenzaldehyde, succinic semialdehyde, 4-nitrobenzaldehyde and 9,10-phenathrenequinone. The two most highly purified rAFAR2-containing preparations eluted from the Cibacron Blue column were 91 and 98% homogeneous. Analysis of these by SDS/PAGE indicated that the least anionic (peak CBA5) comprised a polypeptide of 37.0 kDa, whereas the most anionic (peak CBA6) contained two closely migrating polypeptides of 36.8 and 37.0 kDa; by contrast, in the present study, rAFAR1 was estimated by SDS/PAGE to be composed of 38.0 kDa subunits. Final purification of the 37 kDa polypeptide in CBA5 and CBA6 was accomplished by reversed-phase HPLC. Partial proteolysis of the two preparations of the 37 kDa polypeptide with Staphylococcus aureus V8 protease yielded fragments of identical size, suggesting that they represent the product of a single gene. Furthermore, the peptide maps from CBA5 and CBA6 differed substantially from that yielded by rAFAR1, indicating that they are genetically distinct from the inducible reductase. A peptide generated by CNBr digestion of the 37 kDa polypeptide from CBA6 was shown by Edman degradation to share 88% sequence identity with residues Tyr(168)-Leu(183) of rAFAR1. This provides evidence that the rat protein identified by its cross-reactivity with anti-hAFAR1 serum is an additional member of the AKR7 family. PMID- 10816435 TI - Increased translation efficiency and antizyme-dependent stabilization of ornithine decarboxylase in amino acid-supplemented human colon adenocarcinoma cells, Caco-2. AB - The mechanisms of the response of ornithine decarboxylase(ODC), the rate-limiting enzyme in polyamine biosynthesis, to amino acid supplementation were studied in the human colon adenocarcinoma cell line, Caco-2. Supplementation of serum deprived, subconfluent Caco-2 cells with any one of a series of amino acids (10 mM) resultedin increased ODC activity, reaching a maximum of approx. 12.5-fold after approx. 4 h, over control cells either not supplemented or supplemented with iso-osmolar D-mannitol. Glycine, L-asparagine and L-serine, as well as their D-enantiomers, were the strongest effectors and acted in a concentration dependent manner; millimolar concentrations of most of these amino acids being sufficient to significantly increase ODC activity. In contrast, supplementation with D-methionine, L-lysine, L-aspartate or L-glutamate had little or no effect on ODC activity, whereas supplemental L-methionine, L-arginine, L-ornithine or L cysteine was inhibitory. Polyamine assays showed that the putrescine content of cells varied in accordance with the changes in ODC activity. Western-blot and Northern-blot analyses revealed specifically increased levels of ODC protein but not mRNA,respectively, in response to supplementation with an ODC-inducing amino acid. Suppression of the increase in cycloheximide-treated cellsconfirmed a requirement for protein synthesis. Pulse-labelling of cellswith [(35)S]methionine showed a 3-fold increase in thesynthesis of ODC protein after 4 h of supplementation with glycineor L-serine. Supplemental glycine also augmented, reversibly, the half-life of ODC by almost 4-fold and simultaneously decreased the activity of putrescine-induced free antizyme. These results suggest that translational, but not transcriptional, regulation of ODC takes part in ODC induction by amino acids in Caco-2 cells. However, it also appears to occur in concert with decreased enzyme in activation and/or degradation. PMID- 10816436 TI - Signalling pathways of insulin-like growth factor-I that are augmented by cAMP in FRTL-5 cells. AB - We have reported that pretreatment of rat FRTL-5 thyroid cells with thyrotropin (TSH) markedly potentiates the mitogenic response to insulin-like growth factor-I (IGF-I). The present study was undertaken to determine whether the augmentation by cAMP of IGF-I-dependent tyrosine phosphorylation of known IGF-I receptor substrates plays an important role in the cAMP-dependent potentiation of DNA synthesis induced by IGF-I. Pretreatment with TSH or dibutyryl cAMP did not affect the IGF-I-dependent tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1). In contrast, cAMP pretreatment potentiated the tyrosine phosphorylation of IRS-2 induced by IGF-I, but did not affect the amount of IRS 2. We found that the IGF-I-dependent tyrosine phosphorylation of 66 kDa Shc (Src homology collagen) was markedly increased by cAMP pretreatment, and that this change was mainly due to an increase in the levels of 66 kDa Shc protein. Under these conditions, cAMP pretreatment significantly increased binding of Grb2 (growth-factor-receptor-bound protein 2) to Shc in response to IGF-I, and activation of MAP kinase (mitogen-activated protein kinase) induced by IGF-I was also enhanced by cAMP. The presence of PD98059, an inhibitor of MEK (MAP kinase/Erk kinase), during treatment with IGF-I partially inhibited the cAMP dependent augmentation of DNA synthesis in response to IGF-I. On the other hand, cAMP pretreatment increased binding of the phosphoinositide 3-kinase (PI 3 kinase) p85 subunit to IRS-2, which was reflected in PI 3-kinase activity. LY294002, a PI 3-kinase inhibitor, strongly depressed IGF-I-dependent DNA synthesis after pretreatment with and without TSH or dibutyryl cAMP. Our results suggest that the interaction between cAMP-dependent and IGF-I-dependent pathways leads to an augmentation of cell proliferation, which is mediated, at least in part, through the MAP kinase and PI 3-kinase signalling pathways. These effects are mediated by changes in tyrosine phosphorylation of IGF-I receptor substrates, including IRS-2 and Shc. PMID- 10816437 TI - Phosphocholine-containing, zwitterionic glycosphingolipids of adult Onchocerca volvulus as highly conserved antigenic structures of parasitic nematodes. AB - Human Onchocerca volvulus infection sera were found to recognize zwitterionic glycolipids of O. volvulus and to cross-react with those of other parasitic nematodes (Ascaris suum, Setaria digitata and Litomosoides sigmodontis). By the use of an epitope-specific monoclonal antibody, zwitterionic glycolipids of all these nematode species were observed to contain the antigenic determinant phosphocholine. A hyperimmune serum specific for arthro-series glycolipid structures reacted with the various neutral glycolipids of all these nematodes, which demonstrated that their oligosaccharide moieties belonged to the arthro series of protostomial glycolipids. These results indicated that arthro-series glycosphingolipids carrying, in part, phosphocholine substituents, represent highly conserved, antigenic glycolipid markers of parasitic nematodes. Three glycolipid components of the O. volvulus zwitterionic fraction were structurally characterized by matrix-assisted laser-desorption/ionization time-of-flight MS, methylation analysis and exoglycosidase treatment. Their chemical structures were elucidated to be phosphocholine-6GlcNAc(beta1-3)Man(beta1-4)Glc(1-1)ceramide, GalNAc(beta1-4)[phosphocholine-6]GlcNAc(beta1-3)Man(beta1-4)Glc(1-1) ceramide and Gal(alpha1-3)GalNAc(beta1-4)[phosphocholine-6]GlcNAc(beta1-3)Man(beta 1-4)Glc(1 1)ceramide for the zwitterionic ceramide tri-, tetra- and penta-hexosides respectively. The ceramide composition was found to be dominated by 2 hydroxylated docosanoic (C(22h:0)), tricosanoic (C(23h:0)) and tetracosanoic (C(24h:0)) acids, and C(17) sphingosine (C(d17:1)) (where (h) is hydroxylated and (d) is dihydroxylated). PMID- 10816438 TI - Increase of mitochondria and mitochondrial DNA in response to oxidative stress in human cells. AB - Mitochondrial respiratory function is impaired in the target tissues of patients with mitochondrial diseases and declines with age in various human tissues. It is generally accepted that respiratory-chain defects result in enhanced production of reactive oxygen species and free radicals in mitochondria. Recently, we have demonstrated that the copy number of mitochondrial DNA (mtDNA) is increased in the lung tissues of elderly human subjects. The mtDNA copy number was suggested to be increased by a feedback mechanism that compensates for defects in mitochondria harbouring mutated mtDNA and a defective respiratory system. However, the detailed mechanism remains unclear. In this study, we treated a human lung fibroblast cell line, MRC-5, with H(2)O(2) at concentrations of 90-360 microM. After the treatment for 24-72 h, we found that cells were arrested at G(0) and G(1) phases but that mitochondrial mass and mtDNA content were significantly increased in a concentration- and time-dependent manner. Moreover, the oxidative stress induced by buthionine sulphoximine was also found to cause an increase in mitochondrial mass of the treated cells. Increased uptake of a vital mitochondrial dye Rhodamine 123 and enhanced tetrazolium [MTT, 3-(4, 5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] reduction revealed that the mitochondria increased by H(2)O(2) treatment were functional. In addition, the increase in the mitochondrial mass was also observed in cell-cycle arrested cells induced by mimosine, lovastatin and genistein. Taken together, these findings suggest that the increase in mitochondrial mass and mtDNA content are the early molecular events of human cells in response to endogenous or exogenous oxidative stress through cell-cycle arrest. PMID- 10816439 TI - Perivenous localization of insulin receptor protein in rat liver, and regulation of its expression by glucose and oxygen in hepatocyte cultures. AB - Insulin stimulates glucose utilization in the liver, which occurs mainly in the less aerobic, perivenous, zone. Accordingly, the insulin receptor protein was predominantly expressed in this area, although the insulin receptor mRNA was homogeneously distributed. In hepatocyte cultures venous O(2) partial pressure (pO(2)) induced insulin receptor protein expression. High glucose concentrations enhanced insulin receptor protein under arterial and venous pO(2). The induction of insulin receptor protein by venous pO(2) would explain its zonated expression. PMID- 10816440 TI - Zebrafish ftz-f1 gene has two promoters, is alternatively spliced, and is expressed in digestive organs. AB - Fushi-tarazu Factor-1 (FTZ-F1) is a family of nuclear receptors involved in various developmental processes. We have cloned a zebrafish FTZ-F1 gene, termed ff1, which belongs to the fetoprotein transcription factor/liver receptor homologue-1 (FTF/LRH-1) subgroup of the FTZ-F1 family. Four transcripts arise as a result of differential promoter usage and alternative splicing at the 3'-most exons. The longer transcript, form A, encodes a transcriptional activator. The shorter transcript, form B, lacks the activation domain, and hence could not activate transcription. The difference in promoter usage generates FF1 proteins with different N-terminal sequences. All four transcripts appear to be expressed in most of the adult tissues, whereas, during embryo development, the IIA form is the predominant transcript. Reverse transcriptase-PCR and in situ hybridization experiments showed that the ff1 transcript is expressed in the hypothalamus, spinal cord, mandibular arch and digestive organs, including pancreas, liver, and intestine. The expression of ff1 in the digestive organs implies its function in gut development. PMID- 10816441 TI - Expression and protein-binding studies of the EEN gene family, new interacting partners for dynamin, synaptojanin and huntingtin proteins. AB - EEN, identified initially as a fusion partner to the mixed-lineage leukaemia gene in human leukaemia, and its related members, EEN-B1 and EEN-B2, have recently been shown to interact with two endocytic molecules, dynamin and synaptojanin, as well as with the huntingtin protein. In the present study, we show that the expression of the EEN gene-family members is differentially regulated. Multiple spliced variants were identified for EEN-B2. In the brain, EEN-B1 and EEN-B2 mRNA are preferentially expressed in the cerebellar Purkinje and granule cells, dentate gyrus cells, hippocampal pyramidal neurons and cerebral granule cells. The expression patterns of EEN-B1 and EEN-B2 mRNA in the brain overlap with those of dynamin-I/III, synaptojanin-I and huntingtin, whereas the ubiquitous expression of EEN is consistent with that of dynamin-II. In testes, members of the EEN family are co-expressed with testis-type dynamin and huntingtin in Sertoli cells and germ cells respectively. Our results on the overlapping expression patterns are consistent with the proposed interaction of EEN family members with dynamin, synaptojanin and huntingtin protein in vivo. Although all three EEN family members bind to dynamin and synaptojanin, EEN-B1 has the highest affinity for binding, followed by EEN and EEN-B2. We also demonstrate that amphiphysin, a major synaptojanin-binding protein in brain, can compete with the EEN family for binding to synaptojanin and dynamin. We propose that recruitment of the EEN family by dynamin/synaptojanin to clathrin-coated pits can be regulated by amphiphysin. PMID- 10816442 TI - Characterization of N-myristoyltransferase from Plasmodium falciparum. AB - The gene coding for myristoyl-CoA:protein N-myristoyltransferase (NMT) has been cloned from the malaria parasite Plasmodium falciparum. The gene appears to be single copy and mRNA is expressed in asexual blood-stage forms. Comparison of cDNA and genomic sequences identified three small introns. The open reading frame codes for a 410-amino-acid protein and no evidence of forms with an extended N terminal coding sequence was obtained. Residues important in substrate binding and in the catalytic mechanism in other species are conserved. The protein was expressed from a plasmid in Escherichia coli, partially purified and shown to have enzymic activity using a synthetic peptide substrate. Comparison of the malaria parasite protein with that derived from the human gene showed a different pattern of inhibition by chemical modification. Human NMT activity was inhibited by diethylpyrocarbonate and partially inhibited by iodacetamide, whereas P. falciparum NMT activity was not inhibited by either pre-treatment. Since the enzyme in infectious fungi is a target for potential chemotherapeutic drugs, it should also be investigated in the context of parasitic infections such as that responsible for malaria. PMID- 10816443 TI - Action pattern and substrate specificity of the hyaluronan lyase from group B streptococci. AB - The hyaluronan lyase of group B streptococci rapidly cleaves hyaluronan by an elimination mechanism to yield the unsaturated disaccharide 2-acetamido-2-deoxy-3 O-(beta-D-gluco-4-enepyranosyluronic acid)-D-glucose. Additionally, it has been shown that the enzyme has limited specificity for achondroitin sulphate and cleaves the chain at unsulphated sites [Baker,Yu, Morrison, Averett and Pritchard (1997) Biochem. J. 327,65-71]. In the present extension of that study it was found that 6-sulphated regions of chondroitin sulphate are also susceptible to cleavage by this hyaluronan lyase. Of the four 6- and/or 4-sulphated tetrasaccharides which can be isolated from testicular hyaluronidase digests of chondroitin sulphate, only those two tetrasaccharides with a6-sulphated disaccharide at the reducing end were cleaved. From thisand other data, a model is proposed for the cleavage specificity of hyaluronan lyase on a chondroitin sulphate. Evidence is presented in support of an action pattern for hyaluronan lyase which involves aninitial random endolytic cleavage followed by rapid exolytic and processive release of unsaturated disaccharide. Since the on lyoligosaccharides which tend to accumulate in near-complete digests of hyaluronan are unsaturated, it is argued that the processive cleavage occurs from the non-reducing to the reducing end of a hyaluronan chain. This detailed knowledge of substrate specificity contributes to our understanding of the enzyme's role in Group B streptococcal pathogenesis. In addition, the hyaluronan lyase may find application in sequence studies of chondroitin sulphates. PMID- 10816445 TI - Salmonella enterica serovar typhi uses type IVB pili to enter human intestinal epithelial cells. AB - DNA sequencing upstream of the Salmonella enterica serovar Typhi pilV and rci genes previously identified in the ca. 118-kb major pathogenicity island (X.-L. Zhang, C. Morris, and J. Hackett, Gene 202:139-146, 1997) identified a further 10 pil genes apparently forming a pil operon. The product of the pilS gene, prePilS protein (a putative type IVB structural prepilin) was purified, and an anti prePilS antiserum was raised in mice. Mutants of serovar Typhi either lacking the whole pil operon or with an insertion mutation in the pilS gene were constructed, as was a strain in which the pilN to pilV genes were driven by the tac promoter. The pil(+) strains synthesized type IVB pili, as judged by (i) visualization in the electron microscope of thin pili in culture supernatants of one such strain and (ii) the presence of PilS protein (smaller than the prePilS protein by removal of the leader peptide) on immunoblotting of material pelleted by high speed centrifugation of either the culture supernatant or sonicates of pil(+) strains. Control pil mutants did not express the PilS protein. A pilS mutant of serovar Typhi entered human intestinal INT407 cells in culture to levels only 5 to 25% of those of the wild-type strain, and serovar Typhi entry was strongly inhibited by soluble prePilS protein (50% inhibition of entry at 1.4 microM prePilS). PMID- 10816444 TI - Bcl-2 accelerates retinoic acid-induced growth arrest and recovery in human gastric cancer cells. AB - The role of Bcl-2 as an anti-apoptotic protein has been well documented. In the present work, we present evidence that Bcl-2 may also be involved in cell growth regulation. SC-M1 is an unique cell line which responds to retinoic acid (RA) treatment with reversible growth arrest [Shyu, Jiang, Huang, Chang, Wu, Roffler and Yeh (1995) Eur. J. Cancer 31, 237-243]. In this study, when treated with RA, SC-M1/Bcl2 cells, which were generated by transfecting SC-M1 cells with bcl-2 DNA, were growth-arrested two days earlier than SC-M1/neo cells, which were generated by transfecting SC-M1 cells with vector DNA. This indicates that Bcl-2 accelerates RA-induced growth arrest. In addition to the accelerated growth arrest, RA-treated SC-M1/Bcl2 cells also recovered from growth arrest two days faster than SC-M1/neo cells after the removal of RA. Previously, we had identified the cyclin-dependent kinase inhibitor p21((WAF1/CIP1)) (p21) as a mediator of RA-induced growth arrest [Tsao, Li, Kuo, Liu and Chen (1996) Biochem. J. 317, 707-711]. In a search for the mechanism by which Bcl-2 affects growth regulation, we found that p21 gene expression was more prominent in SC-M1/Bcl2 cells than in SC-M1/neo cells in the presence of RA, but when RA was removed, p21 gene expression levels in SC-M1/Bcl2 cells were also reduced earlier than in SC M1/neo cells. The present report is the first to show that Bcl-2 accelerates not only growth arrest but also recovery from growth arrest. Moreover, the close correlation between the effect of Bcl-2 on both RA-induced growth arrest and RA induced p21 gene expression suggests the possibility that Bcl-2 affects cell growth through the mechanism of p21. PMID- 10816446 TI - Priming with Chlamydia trachomatis major outer membrane protein (MOMP) DNA followed by MOMP ISCOM boosting enhances protection and is associated with increased immunoglobulin A and Th1 cellular immune responses. AB - We previously reported that DNA vaccination was able to elicit cellular immune responses and partial protection against Chlamydia trachomatis infection. However, DNA immunization alone did not generate immune responses or protection as great as that induced by using live organisms. In this study, we evaluated the immunologic effects of a combinational vaccination approach using C. trachomatis mouse pneumonitis (MoPn) major outer membrane protein (MOMP) DNA priming followed by boosting with immune-stimulating complexes (ISCOM) of MOMP protein (MOMP ISCOM) for protection of BALB/c mice against MoPn lung infection. Substantially better protection to challenge infection was observed in mice given combinational vaccination compared with mice given MOMP ISCOM immunization alone, and the protection approximated that induced by live organisms. Enhanced protection was correlated with stronger delayed-type hypersensitivity, higher levels of gamma interferon production, and increased immunoglobulin A antibody responses in lung homogenates. The results indicate that DNA priming followed by ISCOM protein boosting may be useful in designing a fully protective chlamydial vaccine. PMID- 10816448 TI - Protection against virulent Mycobacterium avium infection following DNA vaccination with the 35-kilodalton antigen is accompanied by induction of gamma interferon-secreting CD4(+) T cells. AB - Mycobacterium avium is an opportunistic pathogen that primarily infects immunocompromised individuals, although the frequency of M. avium infection is also increasing in the immunocompetent population. The antigen repertoire of M. avium varies from that of Mycobacterium tuberculosis, with the immunodominant 35 kDa protein being present in M. avium and Mycobacterium leprae but not in members of the M. tuberculosis complex. Here we show that a DNA vector encoding this M. avium 35-kDa antigen (DNA-35) induces protective immunity against virulent M. avium infection, and this protective effect persists over 14 weeks of infection. In C57BL/6 mice, DNA vaccines expressing the 35-kDa protein as a cytoplasmic or secreted protein, both induced strong T-cell gamma interferon (IFN-gamma) and humoral immune responses. Furthermore, the antibody response was to conformational determinants, confirming that the vector-encoded protein had adopted the native conformation. DNA-35 immunization resulted in an increased activated/memory CD4(+) T-cell response, with an accumulation of CD4(+) CD44(hi) CD45RB(lo) T cells and an increase in antigen-specific IFN-gamma production. The protective effect of the DNA-35 vectors against M. avium infection was comparable to that of vaccination with Mycobacterium bovis BCG and significantly greater than that for previous treated infection with M. avium. These results illustrate the importance of the 35-kDa protein in the protective response to M. avium infection and indicate that DNA vaccination successfully promotes a sustained level of protection during chronic M. avium infection. PMID- 10816447 TI - Extending the CD4(+) T-cell epitope specificity of the Th1 immune response to an antigen using a Salmonella enterica serovar typhimurium delivery vehicle. AB - We analyzed the CD4 T-cell immunodominance of the response to a model antigen (Ag), MalE, when delivered by an attenuated strain of Salmonella enterica serovar Typhimurium (SL3261*pMalE). Compared to purified MalE Ag administered with adjuvant, the mapping of the peptide-specific proliferative responses showed qualitative differences when we used the Salmonella vehicle. We observed the disappearance of one out of eight MalE peptides' T-cell reactivity upon SL3261*pMalE immunization, but this phenomenon was probably due to a low level of T-cell priming, since it could be overcome by further immunization. The most striking effect of SL3261*pMalE administration was the activation and stimulation of new MalE peptide-specific T-cell responses that were silent after administration of purified Ag with adjuvant. Ag presentation assays performed with MalE-specific T-cell hybridomas showed that infection of Ag-presenting cells by this intracellular attenuated bacterium did not affect the processing and presentation of the different MalE peptides by major histocompatibility complex (MHC) class II molecules and therefore did not account for immunodominance modulation. Thus, immunodominance of the T-cell response to microorganisms is governed not only by the frequency of the available T-cell repertoire or the processing steps in Ag-presenting cells that lead to MHC presentation but also by other parameters probably related to the infectious process and to the bacterial products. Our results indicate that, upon infection by a microorganism, the specificity of the T-cell response induced against its Ags can be much more effective than with purified Ags and that it cannot completely be mimicked by purified Ags administered with adjuvant. PMID- 10816449 TI - DNA vaccination against tuberculosis: expression of a ubiquitin-conjugated tuberculosis protein enhances antimycobacterial immunity. AB - Genetic immunization is a promising new technology for developing vaccines against tuberculosis that are more effective. In the present study, we evaluated the effects of intracellular turnover of antigens expressed by DNA vaccines on the immune response induced by these vaccines in a mouse model of pulmonary tuberculosis. The mycobacterial culture filtrate protein MPT64 was expressed as a chimeric protein fused to one of three variants of the ubiquitin protein (UbG, UbA, and UbGR) known to differentially affect the intracellular processing of the coexpressed antigens. Immunoblot analysis of cell lysates of in vitro-transfected cells showed substantial differences in the degradation rate of ubiquinated MPT64 (i.e., UbG64 < UbA64 < UbGR64). The specific immune response generated in mice correlated with the stability of the ubiquitin-conjugated antigen. The UbA64 DNA vaccine induced a weak humoral response compared to UbG64, and a mixed population of interleukin-4 (IL-4)- and gamma interferon (IFN-gamma)-secreting cells. Vaccination with the UbGR64 plasmid generated a strong Th1 cell response (high IFN-gamma, low IL-4) in the absence of a detectable humoral response. Aerogenic challenge of vaccinated mice with Mycobacterium tuberculosis indicated that immunization with both the UbA64- and UbGR64-expressing plasmids evoked an enhanced protective response compared to the vector control. The expression of mycobacterial antigens from DNA vaccines as fusion proteins with a destabilizing ubiquitin molecule (UbA or UbGR) shifted the host response toward a stronger Th1 type immunity which was characterized by low specific antibody levels, high numbers of IFN-gamma-secreting cells, and significant resistance to a tuberculous challenge. PMID- 10816450 TI - Adhesion molecule deficiencies increase Porphyromonas gingivalis-induced alveolar bone loss in mice. AB - Alveolar bone resorption can be induced in specific-pathogen-free mice by oral infection with Porphyromonas gingivalis (P. J. Baker, R. T. Evans, and D. C. Roopenian, Arch. Oral Biol. 39:1035-1040, 1994). Here we used a mouse strain, C57BL/6J, which is relatively resistant to P. gingivalis-induced bone loss to examine whether partial or complete deletion of various adhesion molecules would increase susceptibility. Complete deletion of P-selectin or nearly complete lack of expression of intercellular adhesion molecule 1 (ICAM-1) led to increased susceptibility to bone resorption after oral infection, while a hypomorphic defect in beta(2)-integrins did not. Both the total amount of bone lost and the number of sites at which there was significant loss were increased in mice deficient in either ICAM-1 or P-selectin. Each of the three adhesion molecule deficiencies was sufficient to decrease P. gingivalis-specific serum immunoglobulin G responses, but lower antibody titers did not lead to increased bone loss in partially beta(2)-integrin-deficient mice. In conclusion, P-selectin and ICAM-1 deficiencies increase susceptibility to and severity of alveolar bone loss after P. gingivalis infection. This finding underscores the importance of innate immunity in protection against P. gingivalis-induced alveolar bone resorption. PMID- 10816451 TI - Enterohemorrhagic Escherichia coli induces apoptosis which augments bacterial binding and phosphatidylethanolamine exposure on the plasma membrane outer leaflet. AB - Enterohemorrhagic Escherichia coli (EHEC) is a gastrointestinal pathogen that causes watery diarrhea and hemorrhagic colitis and can lead to serious and even fatal complications such as hemolytic uremic syndrome. We investigated the ability of EHEC to kill host cells using three human epithelial cell lines. Analysis of phosphatidylserine expression, internucleosomal cleavage of host cell DNA and morphological changes detected by electron microscopy changes revealed evidence of apoptotic cell death. The rates and extents of cell death were similar for both verotoxin-producing and nonproducing strains of EHEC as well as for a related gastrointestinal pathogen, enteropathogenic E. coli (EPEC). The induction of apoptosis by bacterial attachment was independent of verotoxin production and greater than that produced by a similar treatment with verotoxin alone. Expression of phosphatidylethanolamine, previously reported to bind EHEC and EPEC, was also increased on apoptotic cells but with little correlation to phosphatidylserine expression. Phosphatidylethanolamine levels but not phosphatidylserine levels on dying cells correlated with EHEC binding. Cells treated with phosphatidylethanolamine-containing liposomes also showed increased EHEC binding. These results suggest that bacterial induction of apoptosis offers an advantage for bacterial attachment by augmenting outer leaflet levels of the phosphatidylethanolamine receptor. PMID- 10816452 TI - Effect of preexisting immunity to Salmonella on the immune response to recombinant Salmonella enterica serovar typhimurium expressing a Porphyromonas gingivalis hemagglutinin. AB - Recombinant Salmonella strains expressing foreign heterologous genes have been extensively studied as live oral vaccine delivery vectors. We have investigated the mucosal and systemic immune responses following oral immunization with a recombinant Salmonella enterica serovar Typhimurium expressing the hemagglutinin HagB from Porphyromonas gingivalis, a suspected etiological agent of adult periodontal disease. We have previously shown a primary mucosal and systemic response following oral immunization with chi4072/pDMD1 and recall responses following boosting at 14 weeks after primary immunization. In this study, we examined the effects of earlier boosting as well as the effects of deliberately induced immunity to the Salmonella carrier strain on subsequent immune responses. Mice boosted at week 7 following immunization, a point which corresponded to the peak of the primary response, generally showed lower responses than those boosted at week 14. When mice were preimmunized with the Salmonella carrier alone and then immunized with the recombinant strain 7 or 14 weeks later, significant reductions were seen for serum immunoglobulin G (IgG) antibodies at week 14 and for salivary IgA at week 7. No reductions were seen in serum IgA or vaginal wash IgA antibodies. Mice appear to be refractory to boosting with orally administered salmonellae at 7 weeks. Deliberate immunization with the carrier strain did not appreciably affect recall responses at 14 weeks, with the exception of the serum IgG responses, nor did it affect colonization of the Peyer's patches. PMID- 10816453 TI - Infection of macrophage-like THP-1 cells with Mycobacterium avium results in a decrease in their ability to phosphorylate nucleolin. AB - This study of the phosphorylation ability of macrophage-like cells upon infection with Mycobacterium avium was undertaken to establish potential targets of the interference with host response mechanisms. Cytosolic and membrane fractions from noninfected and infected cells were incubated with [gamma-(32)P]ATP, in the presence of Mg(2+) and the absence of Ca(2+), and the patterns of phosphoproteins synthesized were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Lower levels of a 110-kDa phosphoprotein were observed in association with cytosolic fractions from mycobacterium-infected cells compared to noninfected cells or cells treated with lipopolysaccharide or having ingested Escherichia coli or killed M. avium. The 110-kDa phosphoprotein was present in the soluble fraction (230,000 x g supernatant) after the kinase incubation, from where it was partially purified and identified as phosphonucleolin by amino acid sequencing. The decrease in nucleolin phosphorylation observed was not related to changes in the cytosolic or membrane levels of this protein, and was detected also in the cytosolic fraction of (32)P-labeled intact cells. PMID- 10816454 TI - Mucosal and systemic immune responses to chimeric fimbriae expressed by Salmonella enterica serovar typhimurium vaccine strains. AB - Recombinant live oral vaccines expressing pathogen-derived antigens offer a unique set of attractive properties. Among these are the simplicity of administration, the capacity to induce mucosal and systemic immunity, and the advantage of permitting genetic manipulation for optimal antigen presentation. In this study, the benefit of having a heterologous antigen expressed on the surface of a live vector rather than intracellularly was evaluated. Accordingly, the immune response of mice immunized with a Salmonella enterica serovar Typhimurium vaccine strain expressing the Escherichia coli 987P fimbrial antigen on its surface (Fas(+)) was compared with the expression in the periplasmic compartment (Fas(-)). Orally immunized BALB/c mice showed that 987P fimbriated Salmonella serovar Typhimurium CS3263 (aroA asd) with pCS151 (fas(+) asd(+)) elicited a significantly higher level of 987P-specific systemic immunoglobulin G (IgG) and mucosal IgA than serovar Typhimurium CS3263 with pCS152 (fasD mutant, asd(+)) expressing 987P periplasmic antigen. Further studies were aimed at determining whether the 987P fimbriae expressed by serovar Typhimurium chi4550 (cya crp asd) could be used as carriers of foreign epitopes. For this, the vaccine strain was genetically engineered to express chimeric fimbriae carrying the transmissible gastroenteritis virus (TGEV) C (379-388) and A (521-531) epitopes of the spike protein inserted into the 987P major fimbrial subunit FasA. BALB/c mice administered orally serovar Typhimurium chi4550 expressing the chimeric fimbriae from the tet promoter in pCS154 (fas(+) asd(+)) produced systemic antibodies against both fimbria and the TGEV C epitope but not against the TGEV A epitope. To improve the immunogenicity of the chimeric fimbriae, the in vivo inducible nirB promoter was inserted into pCS154, upstream of the fas genes, to create pCS155. In comparison with the previously used vaccine, BALB/c mice immunized orally with serovar Typhimurium chi4550/pCS155 demonstrated significantly higher levels of serum IgG and mucosal IgA against 987P fimbria. Moreover, mucosal IgA against the TGEV C epitope was only detected with serovar Typhimurium chi4550/pCS155. The induced antibodies also recognized the epitopes in the context of the full-length TGEV spike protein. Hence, immune responses to heterologous chimeric fimbriae on Salmonella vaccine vectors can be optimized by using promoters known to be activated in vivo. PMID- 10816455 TI - Interactions between periodontal bacteria and human oral epithelial cells: Fusobacterium nucleatum adheres to and invades epithelial cells. AB - Bacteria are causative agents of periodontal diseases. Interactions between oral bacteria and gingival epithelial cells are essential aspects of periodontal infections. Using an in vitro tissue culture model, a selected group of gram negative anaerobic bacteria frequently associated with periodontal diseases, including Bacteroides forsythus, Campylobacter curvus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia, were examined for their ability to adhere to and invade primary cultures of human gingival epithelial cells (HGEC). The effects of these bacteria on the production of interleukin-8 (IL-8), a proinflammatory chemokine, were also measured. These studies provided an initial demonstration that F. nucleatum adhered to and invaded HGEC and that this was accompanied by high levels of IL-8 secretion from the epithelial cells. The attachment and invasion characteristics of F. nucleatum were also tested using KB cells, an oral epithelial cell line. The invasion was verified by transmission electron microscopy and with metabolic inhibitors. Invasion appeared to occur via a "zipping" mechanism and required the involvement of actins, microtubules, signal transduction, protein synthesis, and energy metabolism of the epithelial cell, as well as protein synthesis by F. nucleatum. A spontaneous mutant, lam, of F. nucleatum, isolated as defective in autoagglutination, was unable to attach to or invade HGEC or KB cells, further indicating the requirement of bacterial components in these processes. Sugar inhibition assays indicated that lectin-like interactions were involved in the attachment of F. nucleatum to KB cells. Investigation of these new virulence phenotypes should improve our understanding of the role of F. nucleatum in periodontal infections. PMID- 10816456 TI - Microgravity as a novel environmental signal affecting Salmonella enterica serovar Typhimurium virulence. AB - The effects of spaceflight on the infectious disease process have only been studied at the level of the host immune response and indicate a blunting of the immune mechanism in humans and animals. Accordingly, it is necessary to assess potential changes in microbial virulence associated with spaceflight which may impact the probability of in-flight infectious disease. In this study, we investigated the effect of altered gravitational vectors on Salmonella virulence in mice. Salmonella enterica serovar Typhimurium grown under modeled microgravity (MMG) were more virulent and were recovered in higher numbers from the murine spleen and liver following oral infection compared to organisms grown under normal gravity. Furthermore, MMG-grown salmonellae were more resistant to acid stress and macrophage killing and exhibited significant differences in protein synthesis than did normal-gravity-grown cells. Our results indicate that the environment created by simulated microgravity represents a novel environmental regulatory factor of Salmonella virulence. PMID- 10816457 TI - Inhibition of leukocyte entry into the brain by the selectin blocker fucoidin decreases interleukin-1 (IL-1) levels but increases IL-8 levels in cerebrospinal fluid during experimental pneumococcal meningitis in rabbits. AB - The polysaccharide fucoidin is a selectin blocker that inhibits leukocyte recruitment into the cerebrospinal fluid (CSF) during experimental pneumococcal meningitis. In the present study, the effect of fucoidin treatment on the release of the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), and IL-8 into the CSF was investigated. Rabbits (n = 7) were treated intravenously with 10 mg of fucoidin/kg of body weight every second hour starting 4 h after intracisternal inoculation of approximately 10(6) CFU of Streptococcus pneumoniae type 3 (untreated control group, n = 7). CSF samples were obtained every second hour during a 16-h study period. Treatment with fucoidin caused a consistent and significant decrease in CSF IL-1 levels (in picograms per milliliter) between 12 and 16 h (0 versus 170, 0 versus 526, and 60 versus 1,467, respectively; P < 0.02). A less consistent decrease in CSF TNF alpha levels was observed in the fucoidin-treated group, but with no significant difference between the two groups (P > 0.05). In contrast, there was no attenuation in CSF IL-8 levels. Indeed, there was a significant increase in CSF IL-8 levels (in picograms per milliliter) in the fucoidin-treated group at 10 and 12 h (921 versus 574 and 1,397 versus 569, respectively; P < 0.09). In conclusion, our results suggest that blood-derived leukocytes mainly are responsible for the release of IL-1 and to some degree TNF-alpha into the CSF during pneumococcal meningitis, whereas IL-8 may be produced by local cells within the brain. PMID- 10816458 TI - SspA is required for lethal Salmonella enterica serovar Typhimurium infections in calves but is not essential for diarrhea. AB - Salmonella pathogenicity island 1 (SPI-1) encodes virulence determinants, which are important for enteropathogenicity in calves. To determine whether the Salmonella enterica serovar Typhimurium SPI-1 effector proteins SspA and SptP are important for enteropathogenicity, strains lacking these proteins were tested during oral infection of calves. Calves infected with a sptP mutant or its isogenic parent developed diarrhea and lethal morbidity. In contrast, calves infected with an sspA mutant developed diarrhea, which resolved within 10 days but did not result in mortality. The sspA mutant was recovered from bovine intestinal tissues at numbers similar to those obtained for its isogenic parent and caused marked intestinal lesions. Thus, the severity of pathological changes caused by serovar Typhimurium strains or their ability to cause diarrhea were not predictive of their ability to cause lethal morbidity in calves. We conclude that factors other than or in addition to bacterial colonization, intestinal lesions, or electrolyte loss contribute to lethal morbidity in calves infected with serovar Typhimurium. PMID- 10816459 TI - Naturally acquired and vaccine-elicited antibodies block erythrocyte cytoadherence of the Plasmodium vivax Duffy binding protein. AB - Malaria merozoites require the presence of specific surface receptors on the red blood cell for invasion. Plasmodium vivax, requires the Duffy blood group antigen as an obligate receptor for invasion. The parasite Duffy binding protein (DBP) is the ligand involved in this process, making the DBP a potential vaccine candidate. A preliminary objective was to study whether people exposed to vivax malaria acquire antibodies that have the ability to block erythrocyte cytoadherence to the PvDBP. In comparison, we studied the immunogenicity of various recombinant DBP vaccines and investigated their potential to induct antifunctional antibodies. In order to do so, recombinant proteins to different regions of the putative ectodomain of the DBP and a DNA vaccine were used to immunize laboratory animals. An in vitro cytoadherence assay was used to investigate the presence of antifunctional antibodies in plasmas from people naturally exposed to vivax malaria, as well as in antisera obtained by animal vaccination. Our results showed that human plasma from populations naturally exposed to vivax malaria, as well as antisera obtained by vaccination using recombinant proteins, a DNA vaccine, and a synthetic peptide to DBP, inhibited in vitro binding of human erythrocytes to the DBP ligand domain (DBP(II)) in correlation to their previously measured antibody titer. Our results provide further evidence for the vaccine potential of this essential parasite adhesion molecule. PMID- 10816460 TI - Characterization of binding of Candida albicans to small intestinal mucin and its role in adherence to mucosal epithelial cells. AB - In order to approximate and adhere to mucosal epithelial cells, Candida must traverse the overlying mucus layer. Interactions of Candida species with mucin and human buccal epithelial cells (BECs) were thus investigated in vitro. Binding of the Candida species to purified small intestinal mucin showed a close correlation with their hierarchy of virulence. Significant differences (P < 0.05) were found among three categories of Candida species adhering highly (C. dubliniensis, C. tropicalis, and C. albicans), moderately (C. parapsilosis and C. lusitaniae) or weakly (C. krusei and C. glabrata) to mucin. Adherence of C. albicans to BECs was quantitatively inhibited by graded concentrations of mucin. However, inhibition of adherence was reversed by pretreatment of mucin with pronase or C. albicans secretory aspartyl proteinase Sap2p but not with sodium periodate. Saturable concentration- and time-dependent binding of mucin to C. albicans was abrogated by pronase or Sap2p treatment of mucin but was unaffected by beta-mercaptoethanol, sodium periodate, neuraminidase, lectins, or potentially inhibitory sugars. Probing of membrane blots of the mucin with C. albicans revealed binding of the yeast to the 66-kDa cleavage product of the 118-kDa C terminal glycopeptide of mucin. Although no evidence was found for the participation of C. albicans cell surface mannoproteins in specific receptor ligand binding to mucin, inhibition of binding by p-nitrophenol (1 mM) and tetramethylurea (0.36 M) revealed that hydrophobic interactions are involved in adherence of C. albicans to mucin. These results suggest that C. albicans may both adhere to and enzymatically degrade mucins by the action of Saps, and that both properties may act to modulate Candida populations in the oral cavity and gastrointestinal tract. PMID- 10816461 TI - Enterotoxicity and cytotoxicity of Vibrio parahaemolyticus thermostable direct hemolysin in in vitro systems. AB - Vibrio parahaemolyticus is a marine bacterium known to be a common cause of seafood gastroenteritis worldwide. The thermostable direct hemolysin (TDH) has been proposed to be a major virulence factor of V. parahaemolyticus. TDH causes intestinal fluid secretion as well as cytotoxicity in a variety of cell types. In this study, we investigated the interplay between the hemolysin's enterotoxic and cytotoxic effects by using both human and rat cell monolayers. As revealed by microspectrofluorimetry, the toxin causes a dose-dependent increase in intracellular free calcium in both Caco-2 and IEC-6 cells. This effect was reversible only when low toxin concentrations were tested. The TDH-activated ion influx pathway is not selective for calcium but admits ions such sodium and manganese as well. Furthermore, in the same range of concentration, the hemolysin triggers a calcium-dependent chloride secretion. At high concentrations, TDH induces a dose-dependent but calcium-independent cell death as assessed by functional, biochemical, and morphological assays. PMID- 10816462 TI - Identification and expression of a Mycoplasma gallisepticum surface antigen recognized by a monoclonal antibody capable of inhibiting both growth and metabolism. AB - In order to identify antigenic proteins of Mycoplasma gallisepticum, monoclonal antibodies (MAbs) against virulent M. gallisepticum R strain were produced in mice. MAb 35A6 was selected for its abilities to inhibit both growth and metabolism of M. gallisepticum in vitro. The MAb recognized a membrane protein with an apparent molecular mass of 120 kDa. The corresponding gene, designated the mgc3 gene, was cloned from an M. gallisepticum genomic DNA expression library and sequenced. The mgc3 gene is a homologue of the ORF6 gene encoding 130-kDa protein in the P1 operon of M. pneumoniae and is localized downstream of the mgc1 gene, a homologue of the P1 gene. To assess the characteristics of MGC3 protein, all 10 TGA codons in the mgc3 gene, which encode a tryptophan in the Mycoplasma species, were replaced with TGG codons, and recombinant fowlpox viruses (FPV) harboring the altered mgc3 gene were constructed. One of the recombinant FPVs was improved to express MGC3 protein on the cell surface in which the signal peptide of MGC3 protein was replaced with one from Marek's disease virus gB. These results should provide the impetus to develop a vaccine based on MGC3 protein which can induce antibodies with both growth inhibition and metabolic-inhibition activities using a recombinant FPV. PMID- 10816463 TI - Intercellular communication in Helicobacter pylori: luxS is essential for the production of an extracellular signaling molecule. AB - Individual bacteria of numerous species can communicate and coordinate their actions via the production, release, and detection of extracellular signaling molecules. In this study, we used the Vibrio harveyi luminescence bioassay to determine whether Helicobacter pylori produces such a factor. Cell-free conditioned media from H. pylori strains 60190 and 26695 each induced >100-fold greater luminescence in V. harveyi than did sterile culture medium. The H. pylori signaling molecule had a molecular mass of <10 kDa, and its activity was unaffected by heating to 80 degrees C for 5 min or protease treatment. The genome sequence of H. pylori 26695 does not contain any gene predicted to encode an acyl homoserine lactone synthase but does contain an orthologue of luxS, which is required for production of autoinducer-2 (AI-2) in V. harveyi. To evaluate the role of luxS in H. pylori, we constructed luxS null mutants derived from H. pylori 60190 and 26695. Conditioned media from the wild-type H. pylori strains induced >100-fold-greater luminescence in the V. harveyi bioassay than did conditioned medium from either mutant strain. Production of the signaling molecule was restored in an H. pylori luxS null mutant strain by complementation with a single intact copy of luxS placed in a heterologous site on the chromosome. In addition, Escherichia coli DH5alpha produced autoinducer activity following the introduction of an intact copy of luxS from H. pylori. Production of the signaling molecule by H. pylori was growth phase dependent, with maximal production occurring in the mid-exponential phase of growth. Transcription of H. pylori vacA also was growth phase dependent, but this phenomenon was not dependent on luxS activity. These data indicate that H. pylori produces an extracellular signaling molecule related to AI-2 from V. harveyi. We speculate that this signaling molecule may play a role in regulating H. pylori gene expression. PMID- 10816464 TI - Identification of an immunodominant ABC transporter in methicillin-resistant Staphylococcus aureus infections. AB - Immunoblotting sera from 26 patients with septicemia due to an epidemic strain of methicillin-resistant Staphylococcus aureus (EMRSA-15), 6 of whom died, revealed an immunodominant EMRSA-15 antigen at 61 kDa. There was a statistically significant correlate (P < 0.001) between survival and immunoglobulin G to the 61 kDa band. The antigen was identified by sequencing positive clones obtained by screening a genomic expression library of EMRSA-15 with pooled sera from patients taken after the septicemic episode. Eluted antibody reacted with the 61-kDa antigen on immunoblots. The amino terminus was obtained by searching the S. aureus NCTC 8325 and MRSA strain COL databases, and the whole protein was expressed in Escherichia coli TOP 10F'. The derived amino acid sequence showed homology with ABC transporters, with paired Walker A and Walker B motifs and 73% homology to YkpA from Bacillus subtilis. Epitope mapping of the derived amino acid sequence with sera from patients who had recovered from EMRSA-15 septicemia delineated seven epitopes. Three of these epitopes, represented by peptides 1 (KIKVYVGNYDFWYQS), 2 (TVIVVSHDRHFLYNNV), and 3 (TETFLRGFLGRMLFS), were synthesized and used to isolate human recombinant antibodies from a phage antibody display library. Recombinant antibodies against peptides 1 and 2 gave logarithmic reductions in organ colony counts, compared with control groups, in a mouse model of the infection. This study suggests the potential role of an ABC transporter as a target for immunotherapy. PMID- 10816465 TI - Brucella abortus and its closest phylogenetic relative, Ochrobactrum spp., differ in outer membrane permeability and cationic peptide resistance. AB - The outer membrane (OM) of the intracellular parasite Brucella abortus is permeable to hydrophobic probes and resistant to destabilization by polycationic peptides and EDTA. The significance of these unusual properties was investigated in a comparative study with the opportunistic pathogens of the genus Ochrobactrum, the closest known Brucella relative. Ochrobactrum spp. OMs were impermeable to hydrophobic probes and sensitive to polymyxin B but resistant to EDTA. These properties were traced to lipopolysaccharide (LPS) because (i) insertion of B. abortus LPS, but not of Escherichia coli LPS, into Ochrobactrum OM increased its permeability; (ii) permeability and polymyxin B binding measured with LPS aggregates paralleled the results with live bacteria; and (iii) the predicted intermediate results were obtained with B. abortus-Ochrobactrum anthropi and E. coli-O. anthropi LPS hybrid aggregates. Although Ochrobactrum was sensitive to polymyxin, self-promoted uptake and bacterial lysis occurred without OM morphological changes, suggesting an unusual OM structural rigidity. Ochrobactrum and B. abortus LPSs showed no differences in phosphate, qualitative fatty acid composition, or acyl chain fluidity. However, Ochrobactrum LPS, but not B. abortus LPS, contained galacturonic acid. B. abortus and Ochrobactrum smooth LPS aggregates had similar size and zeta potential (-12 to -15 mV). Upon saturation with polymyxin, zeta potential became positive (1 mV) for Ochrobactrum smooth LPS while remaining negative (-5 mV) for B. abortus smooth LPS, suggesting hindered access to inner targets. These results show that although Ochrobactrum and Brucella share a basic OM pattern, subtle modifications in LPS core cause markedly different OM properties, possibly reflecting the adaptive evolution of B. abortus to pathogenicity. PMID- 10816466 TI - Treponema pallidum subsp. pertenue displays pathogenic properties different from those of T. pallidum subsp. pallidum. AB - The present study described the susceptibility of C4D guinea pigs to cutaneous infection with Treponema pallidum subsp. pertenue Haiti B strain. The general manifestations of the disease in adults and neonates differ, to a certain degree, from those induced by T. pallidum subsp. pallidum Nichols strain. Noticeable differences between the infections were reflected in the character of the skin lesions, their onset and persistence, and the kinetics of the humoral response. The incidence and dissemination of cutaneous yaws lesions in very young guinea pigs were remarkably different from the low frequency observed in a similar age group of syphilis infection, 100 versus 17%, respectively. Moreover, as opposed to T. pallidum subsp. pallidum, T. pallidum subsp. pertenue does not cross the placenta. Offspring born to yaws-infected mothers did not produce immunoglobulin M antibodies and their organs, examined by PCR and rabbit infectivity test (RIT), were all negative. Examination of a large number of tissues and organs in adult, neonate, and maternal yaws by PCR and RIT clearly demonstrated that, unlike syphilis, there was a low incidence and short persistence of the yaws pathogen in internal organs. These findings stress the dermotropic rather than the organotropic character of yaws and provide further evidence of distinctive biological and pathological differences between yaws and venereal syphilis. PMID- 10816467 TI - Streptococcal erythrogenic toxin B abrogates fibronectin-dependent internalization of Streptococcus pyogenes by cultured mammalian cells. AB - Streptococcus pyogenes secretes several proteins that influence host-pathogen interactions. A tissue-culture model was used to study the influence of the secreted cysteine protease streptococcal erythrogenic toxin B (SPE B) on the interaction between S. pyogenes strain NZ131 (serotype M49) and mammalian cells. Inactivation of the speB gene enhanced fibronectin-dependent uptake of the pathogen by Chinese hamster ovary (CHO-K1) cells compared to that in the isogenic wild-type strain. Preincubation of the NZ131 speB mutant with purified SPE B protease significantly inhibited fibronectin-dependent uptake by both CHO-K1 and CHO-pgs745 cells. The effect was attributed to an abrogation of fibronectin binding to the surface of the bacteria that did not involve either the M49 protein or the streptococcal fibronectin-binding protein SfbI. In contrast, pretreatment of the NZ131 speB mutant with SPE B did not influence sulfated polysaccharide-mediated uptake by CHO-pgs745 cells. The results indicate that the SPE B protease specifically alters bacterial cell surface proteins and thereby influences pathogen uptake. PMID- 10816468 TI - A major secreted elastase is essential for pathogenicity of Aeromonas hydrophila. AB - Aeromonas hydrophila is an opportunistic pathogen and the leading cause of fatal hemorrhagic septicemia in rainbow trout. A gene encoding an elastolytic activity, ahyB, was cloned from Aeromonas hydrophila AG2 into pUC18 and expressed in Escherichia coli and in the nonproteolytic species Aeromonas salmonicida subsp. masoucida. Nucleotide sequence analysis of the ahyB gene revealed an open reading frame of 1,764 nucleotides with coding capacity for a 588-amino-acid protein with a molecular weight of 62,728. The first 13 N-terminal amino acids of the purified protease completely match those deduced from DNA sequence starting at AAG (Lys 184). This finding indicated that AhyB is synthesized as a preproprotein with a 19-amino-acid signal peptide, a 164-amino-acid N-terminal propeptide, and a 405 amino-acid intermediate which is further processed into a mature protease and a C terminal propeptide. The protease hydrolyzed casein and elastin and showed a high sequence similarity to other metalloproteases, especially with the mature form of the Pseudomonas aeruginosa elastase (52% identity), Helicobacter pylori zinc metalloprotease (61% identity), or proteases from several species of Vibrio (52 to 53% identity). The gene ahyB was insertionally inactivated, and the construct was used to create an isogenic ahyB mutant of A. hydrophila. These first reports of a defined mutation in an extracellular protease of A. hydrophila demonstrate an important role in pathogenesis. PMID- 10816469 TI - Listeriolysin O as a reporter to identify constitutive and in vivo-inducible promoters in the pathogen Listeria monocytogenes. AB - Listeria monocytogenes is a facultative intracellular gram-positive bacterium capable of growing in the cytoplasm of infected host cells. Bacterial escape from the phagosomal vacuole of infected cells is mainly mediated by the pore-forming hemolysin listeriolysin O (LLO) encoded by hly. LLO-negative mutants of L. monocytogenes are avirulent in the mouse model. We have developed a genetic system with hly as a reporter gene allowing the identification of both constitutive and in vivo-inducible promoters of this pathogen. Genomic libraries were created by randomly inserting L. monocytogenes chromosomal fragments upstream of the promoterless hly gene cloned into gram-positive and gram-negative shuttle vectors and expressed in an LLO-negative mutant strain. With this hly based promoter trap system, combined with access to the L. monocytogenes genome database, we identified 20 in vitro-transcribed genes, including genes encoding (i) p60, a previously known virulence gene, (ii) a putative new hemolysin, and (iii) two proteins of the general protein secretion pathway. By using the hly based system as an in vivo expression technology tool, nine in vivo-induced loci of L. monocytogenes were identified, including genes encoding (i) the previously known in vivo-inducible phosphatidylinositol phospholipase C and (ii) a putative N-acetylglucosamine epimerase, possibly involved in teichoic acid biosynthesis. The use of hly as a reporter is a simple and powerful alternative to classical methods for transcriptional analysis to monitor promoter activity in L. monocytogenes. PMID- 10816470 TI - Heterologous expression of an immunogenic pneumococcal type 3 capsular polysaccharide in Lactococcus lactis. AB - In order to develop a new system for the analysis of capsular biosynthetic pathways we have explored the possibility of expressing type 3 capsular polysaccharide (CPS) from the pathogen Streptococcus pneumoniae in Lactococcus lactis, an unencapsulated lactic acid bacterium being developed as a vaccine delivery vehicle for mucosal immunization. Only three of the four type 3 CPS biosynthesis genes were found to be necessary for the abundant formation (120 mg liter(-1)) of an extracellular type 3 CPS in L. lactis, implying a role for the type 3-specific synthase in the extracellular transport of the CPS or implying the existence of an alternative export system in L. lactis. The authenticity of the expressed heterologous polysaccharide was established by chemical and immunological analyses. Proton and carbon nuclear magnetic resonance spectroscopy of CPSs purified from L. lactis and S. pneumoniae showed that the two CPS structures were identical. When mice were immunized intraperitoneally with 3.5 x 10(6) CFU of live recombinant lactococci expressing a total of approximately 0.5 microgram of type 3 CPS, the immune responses elicited appeared identical to those observed in mice inoculated with 0.5 microgram of type 3 CPS purified from S. pneumoniae. These findings show that L. lactis is a useful host in which to study the role and function of genes involved in the production of bacterial capsules. Additionally, L. lactis shows potential as a host for the safe production of capsule antigens and as a vaccine delivery vehicle for polysaccharide antigens. PMID- 10816471 TI - Inhibition of staphylococcal enterotoxin B-induced lymphocyte proliferation and tumor necrosis factor alpha secretion by MAb5, an anti-toxic shock syndrome toxin 1 monoclonal antibody. AB - Toxic shock syndrome (TSS) is primarily caused by toxic shock syndrome toxin-1 (TSST-1) and staphylococcal enterotoxin B (SEB). These toxins belong to a family of pyrogenic toxin superantigens (PTSAgs) produced by Staphylococcus aureus and exhibit several shared biological properties, including the induction of massive cytokine release and Vbeta-specific T-cell proliferation. The crystal structures of most PTSAgs are now published, and they demonstrate a striking similarity in conformational architecture even though their primary protein sequences are different. Despite these structural and immunobiological similarities, no cross reactivity between TSST-1 and other PTSAgs has been demonstrated in serological or neutralization assays. Our laboratory has developed a neutralizing murine anti TSST-1 monoclonal antibody (MAb5) which displayed cross-reactivity with SEB by enzyme-linked immunosorbent assay. The aim of the present study was to evaluate whether MAb5 can also cross-neutralize SEB-induced superantigenic activities in vitro. MAb5 was found to partially inhibit SEB-induced T-cell mitogenesis (63%) and tumor necrosis factor alpha (TNF-alpha) secretion (70%) in human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner, while an isotypic anti TSST-1 monoclonal antibody showed no effect. Epitope mapping revealed that MAb5 bound to TSST-1 residues 47 to 56 ((47)FPSPYYSPAF(56)) and to SEB residues 83 to 92 ((83)DVFGANYYYQ(92)), sequences that located in different regions of these toxins and are structurally dissimilar. SEB peptide (83)DVFGANYYYQ(92) was synthesized and found to also inhibit SEB-induced mitogenesis and TNF-alpha secretion in human PBMC. Our results demonstrate for the first time that MAb5 binds to different epitopes on TSST-1 and SEB that appear functionally important in inducing T-cell mitogenesis and TNF-alpha secretion in vitro. PMID- 10816472 TI - Identification and characterization of murine cytotoxic T cells that kill Mycobacterium tuberculosis. AB - As we seek to develop and evaluate new vaccines against tuberculosis, it is desirable that we understand the mechanisms of protective immunity in our models. Adoptive transfer of protection with hsp65-specific T-cell clones from infected or vaccinated mice into naive mice had indicated that cytotoxic T cells can make a major contribution to protection. We characterized 28 CD4(+) CD8(-) and 28 CD4( ) CD8(+) hsp65-specific T-cell clones derived from infected or vaccinated mice. Half of the CD4(+) CD8(-) and 64% of the CD4(-) CD8(+) clones were cytotoxic. Cytotoxicity was associated with high expression of CD44 and gamma interferon production. Most (86%) of the cytotoxic CD4(+) CD8(-) clones lysed target cells via the Fas-FasL pathway, and most (83%) of the cytotoxic CD4(-) CD8(+) clones lysed target cells via cytotoxic granules. Only the clones using the granule mediated pathway caused substantial loss of viability of virulent Mycobacterium tuberculosis during lysis of infected macrophages, and the degree of killing closely correlated with the availability of granule marker enzyme activity. Granule-mediated cytotoxicity thus may have a key role in protection against tuberculosis by delivering mycobactericidal granule contents. PMID- 10816473 TI - Interaction of Listeria monocytogenes with human brain microvascular endothelial cells: an electron microscopic study. AB - Internalization of Listeria monocytogenes into human brain microvascular endothelial cells (HBMEC) has recently been demonstrated to be dependent upon the inlB gene. In the present scanning electron microscopic study we show that L. monocytogenes efficiently interacts with the surface of HBMEC in an inlB independent manner which is also different from invasion. The inlB-dependent invasion of HBMEC by L. monocytogenes is accompanied by intracellular multiplication, movement, and production of bacterium-containing protrusions. These protrusions extend from the cell surface without perturbation of any adjacent cellular membrane. PMID- 10816474 TI - CS22, a novel human enterotoxigenic Escherichia coli adhesin, is related to CS15. AB - Enterotoxigenic Escherichia coli (ETEC) expresses a broad spectrum of O:H antigens. Serogroup O20 is one of the most prevalent among the ETEC strains lacking any of the defined colonization factors (CFs), in Argentina. An O20:H- strain, ARG-3, adhered to Caco-2 cells and exhibited a thermoregulated 15.7-kDa protein band upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). An antiserum against this protein inhibited ARG-3 adhesion to Caco-2 cells and bound to very thin fibrilla-like structures on the bacterial surface. A 15.7 kDa protein-defective mutant failed to adhere to Caco-2 cells and lacked immunogold-labeled surface structures. The N-terminal amino acid sequence of the structural subunit showed 95% homology to that of CS15 of ETEC (former antigen 8786) and 65% homology with fimbria SEF14 of Salmonella enterica serovar Enteritidis. Nevertheless, the molecular size of ARG-3 adhesin was different from that of CS15, as revealed by SDS-PAGE and mass spectrometry. Both proteins are immunologically related, yet not identical, since an antiserum against the 15.7 kDa protein reacted solely with ARG-3 after absorption with bacteria bearing CS15. Moreover, only under low stringency conditions could DNA from strain ARG-3 be amplified by PCR using primers derived from the nfaA sequence of CS15. Thus, from the DNA sequence obtained from the ARG-3 PCR product, it could be deduced that the subunit protein differed in 30 residues from that of CS15. ARG-3 adhesin was found in 60% of the O20:H- CF-negative ETEC strains from Argentina; however, it appeared restricted to this serotype. We propose the designation CS22 for the herein identified nonfimbrial adhesin of human ETEC. PMID- 10816475 TI - Overexpression of protective antigen as a novel approach to enhance vaccine efficacy of Brucella abortus strain RB51. AB - Brucella abortus strain RB51 is an attenuated rough strain that is currently being used as the official live vaccine for bovine brucellosis in the United States and several other countries. We reasoned that overexpression of a protective antigen(s) of B. abortus in strain RB51 should enhance its vaccine efficacy. To test this hypothesis, we overexpressed Cu/Zn superoxide dismutase (SOD) protein of B. abortus in strain RB51. This was accomplished by transforming strain RB51 with a broad-host-range plasmid, pBBR1MCS, containing the sodC gene along with its promoter. Strain RB51 overexpressing SOD (RB51SOD) was tested in BALB/c mice for its ability to protect against challenge infection with virulent strain 2308. Mice vaccinated with RB51SOD, but not RB51, developed antibodies and cell-mediated immune responses to Cu/Zn SOD. Strain RB51SOD vaccinated mice developed significantly (P < 0.05) more resistance to challenge than those vaccinated with strain RB51 alone. The presence of the plasmid alone in strain RB51 did not alter its vaccine efficacy. Also, overexpression of SOD did not alter the attenuation characteristic of strain RB51. PMID- 10816476 TI - Brucella abortus strain RB51 as a vector for heterologous protein expression and induction of specific Th1 type immune responses. AB - Brucella abortus strain RB51 is a stable, rough, attenuated mutant widely used as a live vaccine for bovine brucellosis. Our ultimate goal is to develop strain RB51 as a preferential vector for the delivery of protective antigens of other intracellular pathogens to which the induction of a strong Th1 type of immune response is needed for effective protection. As a first step in that direction, we studied the expression of a foreign reporter protein, beta-galactosidase of Escherichia coli, and the 65-kDa heat shock protein (HSP65) of Mycobacterium bovis in strain RB51. We cloned the promoter sequences of Brucella sodC and groE genes in pBBR1MCS to generate plasmids pBBSODpro and pBBgroE, respectively. The genes for beta-galactosidase (lacZ) and HSP65 were cloned in these plasmids and used to transform strain RB51. An enzyme assay in the recombinant RB51 strains indicated that the level of beta-galactosidase expression is higher under the groE promoter than under the sodC promoter. In strain RB51 containing pBBgroE/lacZ, but not pBBSODpro/lacZ, increased levels of beta-galactosidase expression were observed after subjecting the bacteria to heat shock or following internalization into macrophage-like J774A.1 cells. Mice vaccinated with either of the beta-galactosidase-expressing recombinant RB51 strains developed specific antibodies of predominantly the immunoglobulin G2a (IgG2a) isotype, and in vitro stimulation of their splenocytes with beta-galactosidase induced the secretion of gamma interferon (IFN-gamma), but not interleukin-4 (IL-4). A Th1 type of immune response to HSP65, as indicated by the presence of specific serum IgG2a, but not IgG1, antibodies, and IFN-gamma, but not IL-4, secretion by the specific-antigen stimulated splenocytes, was also detected in mice vaccinated with strain RB51 containing pBBgroE/hsp65. Studies with mice indicated that expression of beta galactosidase or HSP65 did not alter either the attenuation characteristics of strain RB51 or its vaccine efficacy against B. abortus 2308 challenge. PMID- 10816477 TI - Local anticandidal immune responses in a rat model of vaginal infection by and protection against Candida albicans. AB - Humoral (antibody [Ab]) and cellular Candida-specific immune responses in the vaginas of pseudoestrus rats were investigated during three successive infections by Candida albicans. After the first, protective infection, Abs against mannan and aspartyl proteinase antigens were present in the vaginal fluid, and their titers clearly increased during the two subsequent, rapidly healing infections. In all animals, about 65 and 10% of vaginal lymphocytes (VL) were CD3(+) (T cells) and CD3(-) CD5(+) (B cells), respectively. Two-thirds of the CD3(+) T cells expressed the alpha/beta and one-third expressed the gamma/delta T-cell receptor (TCR). This proportion slightly fluctuated during the three rounds of C. albicans infection, but no significant differences between infected and noninfected rats were found. More relevant were the changes in the CD4(+)/CD8(+) T-cell ratio, particularly for cells bearing the CD25 (interleukin-2 receptor alpha) marker. In fact, a progressively increased number of both CD4(+) alpha/beta TCR and CD4(+) CD25(+) VL was observed after the second and third Candida challenges, reversing the high initial CD8(+) cell number of controls (estrogenized but uninfected rats). The CD3(-) CD5(+) cells also almost doubled from the first to the third infection. Analysis of the cytokines secreted in the vaginal fluid of Candida-infected rats showed high levels of interleukin 12 (IL 12) during the first infection, followed by progressively increasing amounts of IL-2 and gamma interferon during the subsequent infections. No IL-4 or IL-5 was ever detected. During the third infection, VL with in vitro proliferative activity in response to an immunodominant mannoprotein antigen of C. albicans were present in the vaginal tissue. No response to this antigen by mitogen responsive blood, lymph node, and spleen cells was found. In summary, the presence of protective Ab and T helper type 1 cytokines in the vaginal fluids, the in vitro proliferation of vaginal lymphocytes in response to Candida antigenic stimulation, and the increased number of activated CD4(+) cells and some special B lymphocytes after C. albicans challenge constitute good evidence for induction of locally expressed Candida-specific Ab and cellular responses which are potentially involved in anticandidal protection at the vaginal level. PMID- 10816478 TI - Characterization of FimY as a coactivator of type 1 fimbrial expression in Salmonella enterica serovar Typhimurium. AB - Type 1 fimbriae of Salmonella enterica serovar Typhimurium are surface appendages that carry adhesins specific for mannosylated host glycoconjugates. Regulation of the major fimbrial subunit is thought to be controlled by a number of ancillary fim genes, including fimZ, fimY, fimW, and fimU. Previous studies using a FimZ mutant have indicated that this protein is necessary for fimA expression, and in vitro DNA binding assays determined that FimZ is a transcriptional activator that binds directly to the fimA promoter. To determine the role of FimY as a potential regulator of fimbrial expression, a fimY mutant of serovar Typhimurium was generated by allelic exchange. This mutant was found to be phenotypically nonfimbriate. No transcription from the fimA promoter was detected in a fimY mutant containing a fimA-lacZ reporter construct located on the chromosome. In addition, transcription from the cloned fimY promoter was not detected in Escherichia coli unless both FimZ and FimY were present, indicating that these proteins also act as coactivators of fimY expression. Consistent with these results, there is no transcription from a fimY-lacZ reporter construct within a serovar Typhimurium fimY or fimZ mutant. Studies using the fimY-lacZ construct reveal that expression of this gene varies with environmental conditions in a manner similar to fimA expression. Extensive in vitro DNA binding assays using extracts from E. coli that overexpress FimY, as well as partially purified FimY, were unable to identify a specific interaction between FimY and the fimA or fimY promoter. The results indicate that FimY is a positive regulator of fimbrial expression and that this protein acts in cooperation with FimZ to regulate the expression of Salmonella type 1 fimbrial appendages. PMID- 10816479 TI - Antigenic equivalence of human T-cell responses to Mycobacterium tuberculosis specific RD1-encoded protein antigens ESAT-6 and culture filtrate protein 10 and to mixtures of synthetic peptides. AB - The early secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10) are promising antigens for reliable immunodiagnosis of tuberculosis. Both antigens are encoded by RD1, a genomic region present in all strains of Mycobacterium tuberculosis and M. bovis but lacking in all M. bovis bacillus Calmette-Guerin vaccine strains. Production and purification of recombinant antigens are laborious and costly, precluding rapid and large-scale testing. Aiming to develop alternative diagnostic reagents, we have investigated whether recombinant ESAT-6 (rESAT-6) and recombinant CFP-10 (rCFP-10) can be replaced with corresponding mixtures of overlapping peptides spanning the complete amino acid sequence of each antigen. Proliferation of M. tuberculosis specific human T-cell lines in response to rESAT-6 and rCFP-10 and that in response to the corresponding peptide mixtures were almost completely correlated (r = 0.96, P < 0.0001 for ESAT-6; r = 0.98, P < 0.0001 for CFP-10). More importantly, the same was found when gamma interferon production by peripheral blood mononuclear cells in response to these stimuli was analyzed (r = 0.89, P < 0.0001 for ESAT-6; r = 0.89, P < 0.0001 for CFP-10). Whole protein antigens and the peptide mixtures resulted in identical sensitivity and specificity for detection of infection with M. tuberculosis. The peptides in each mixture contributing to the overall response varied between individuals with different HLA-DR types. Interestingly, responses to CFP-10 were significantly higher in the presence of HLA-DR15, which is the major subtype of DR2. These results show that mixtures of synthetic overlapping peptides have potency equivalent to that of whole ESAT-6 and CFP-10 for sensitive and specific detection of infection with M. tuberculosis, and peptides have the advantage of faster production at lower cost. PMID- 10816480 TI - Interleukin-6 induces early gamma interferon production in the infected lung but is not required for generation of specific immunity to Mycobacterium tuberculosis infection. AB - Immunity to Mycobacterium tuberculosis is dependent upon the generation of a protective gamma interferon (IFN-gamma)-producing T-cell response. Recent studies have suggested that interleukin-6 (IL-6) is required for the induction of a protective T-cell response and that IL-4 may suppress the induction of IFN-gamma. To evaluate the role of the cytokines IL-6 and IL-4 in the generation of pulmonary immunity to M. tuberculosis, IL-6 and IL-4 knockout mice were infected with M. tuberculosis via aerosol. The absence of IL-6 led to an early increase in bacterial load with a concurrent delay in the induction of IFN-gamma. However, mice were able to contain and control bacterial growth and developed a protective memory response to secondary infection. This demonstrates that while IL-6 is involved in stimulating early IFN-gamma production, it is not essential for the development of protective immunity against M. tuberculosis. In contrast, while the absence of IL-4 resulted in increased IFN-gamma production, this had no significant effect upon bacterial growth. PMID- 10816481 TI - Evidence of commonality between canine and human extraintestinal pathogenic Escherichia coli strains that express papG allele III. AB - Although dogs have been proposed as carriers of extraintestinal pathogenic Escherichia coli (ExPEC) with infectious potential for humans, presumed host species-specific differences between canine and human ExPEC strains have cast doubt on this hypothesis. The recent discovery that allele III of papG (the P fimbrial adhesin gene) predominates among human cystitis isolates and confers an adherence phenotype resembling that of canine ExPEC prompted the present reevaluation of the canine-human ExPEC connection. Sixteen paired pap-positive urine and rectal E. coli isolates from dogs with urinary tract infection were studied. papG (adhesin) and papA (pilin) allele type, agglutination phenotypes, virulence factor genotypes, and randomly amplified polymorphic DNA and pulsed field gel electrophoresis fingerprints were analyzed and compared with those of human ExPEC controls. The 16 canine strains contained predominantly papG allele III. Agglutination phenotypes segregated strictly according to papG allele status and were homogeneous among strains with the same papG allele profile irrespective of their human versus canine origin. Canine and human PapG variant III peptide sequences were highly homologous, without host species-specific differences. The most prevalent canine papA allele was F48, a novel variant recently identified among human urosepsis isolates. In addition to pap, human ExPEC-associated virulence genes detected among the canine strains included sfa/focDE, sfaS, fyuA, hlyA, cnf1, cdtB, kpsMT-II and -III, rfc, traT, ompT, and a marker for a pathogenicity-associated island from archetypal human ExPEC strain CFT073. Molecular fingerprinting confirmed the fecal origin of all but one canine urine isolate and showed one pair of O6 canine urine and fecal isolates to be extremely similar to an O6 human urosepsis isolate with which they shared all other genotypic and phenotypic characteristics analyzed. These data demonstrate that canine ExPEC strains are similar to, and in some instances essentially indistinguishable from, human ExPEC strains, which implicates dogs and their feces as potential reservoirs of E. coli with infectious potential for humans. PMID- 10816482 TI - Immune response to an 18-kilodalton outer membrane antigen identifies lipoprotein 20 as a Helicobacter pylori vaccine candidate. AB - Experiments were performed using the standardized murine model of Helicobacter pylori infection to determine the immunogenicity of H. pylori outer membrane vesicles in immune protection. These vesicles, which are naturally shed from the surface of the bacterium, induce a protective response when administered intragastrically to mice in the presence of cholera holotoxin, despite the absence of the urease enzyme and associated Hsp54 chaperonin. Immunoblotting identified a specific serum immunoglobulin G (IgG) response to an 18-kDa outer membrane protein in a significant number of immunized animals. This commonly expressed, immunodominant protein was subsequently identified as lipoprotein 20 (Lpp20). Hybridoma backpacks secreting an IgG1 subclass monoclonal antibody to Lpp20 were generated in H. pylori-infected mice and were found to significantly reduce bacterial numbers, providing evidence that this surface-exposed antigen is a true vaccine candidate and not merely an antigenic marker for successful, protective immunization. PMID- 10816483 TI - Antibody is required for protection against virulent but not attenuated Salmonella enterica serovar typhimurium. AB - Resolution of infection with attenuated Salmonella is an active process that requires CD4(+) T cells. Here, we demonstrate that costimulation via the surface molecule CD28, but not antibody production by B cells, is required for clearance of attenuated aroA Salmonella enterica serovar typhimurium. In contrast, specific antibody is critical for vaccine-induced protection against virulent bacteria. Therefore, CD28(+) CD4(+) T cells are sufficient for clearance of avirulent Salmonella in naive hosts, whereas CD4(+) T cells and specific antibodies are required for protection from virulent Salmonella in immune hosts. PMID- 10816484 TI - A model of infected burn wounds using Escherichia coli O18:K1:H7 for the study of gram-negative bacteremia and sepsis. AB - A difficulty that has emerged in the development and preclinical evaluation of adjuvant therapies for gram-negative sepsis is the lack of easily studied animal models that closely mimic human infection. An objective of this study was to adapt a previously described model of infection in burned mice to rats with a defined bacterial strain of Escherichia coli. Challenge with two colonies of live E. coli O18:K1:H7 bacteria into an 8% full-thickness burn of the dorsal skin surface of rats produced predictable bacteremia at 24 to 48 h and 80 to 100% mortality at 3 to 4 days. E. coli O18:K1:H7 was approximately 10-million-fold more virulent than several other gram-negative bacterial strains. The model should be a useful tool in studying the pathogenicity of burn wound infections and in evaluating the efficacy of novel adjuvant therapies for gram-negative sepsis. PMID- 10816485 TI - Construction and characterization of Haemophilus ducreyi lipooligosaccharide (LOS) mutants defective in expression of heptosyltransferase III and beta1,4 glucosyltransferase: identification of LOS glycoforms containing lactosamine repeats. AB - To begin to understand the role of the lipooligosaccharide (LOS) molecule in chancroid infections, we constructed mutants defective in expression of glycosyltransferase genes. Pyocin lysis and immunoscreening was used to identify a LOS mutant of Haemophilus ducreyi 35000. This mutant, HD35000R, produced a LOS molecule that lacked the monoclonal antibody 3F11 epitope and migrated with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Structural studies indicated that the principal LOS glycoform contains lipid A, Kdo, and two of the three core heptose residues. HD35000R was transformed with a plasmid library of H. ducreyi 35000 DNA, and a clone producing the wild-type LOS was identified. Sequence analysis of the plasmid insert revealed one open reading frame (ORF) that encodes a protein with homology to the WaaQ (heptosyltransferase III) of Escherichia coli. A second ORF had homology to the LgtF (glucosyltransferase) of Neisseria meningitidis. Individual isogenic mutants lacking expression of the putative H. ducreyi heptosyltransferase III, the putative glucosyltransferase, and both glycosyltransferases were constructed and characterized. Each mutant was complemented with the representative wild-type genes in trans to restore expression of parental LOS and confirm the function of each enzyme. Matrix-assisted laser desorption ionization mass spectrometry and SDS-PAGE analysis identified several unique LOS glycoforms containing di-, tri-, and poly-N-acetyllactosamine repeats added to the terminal region of the main LOS branch synthesized by the heptosyltransferase III mutant. These novel H. ducreyi mutants provide important tools for studying the regulation of LOS assembly and biosynthesis. PMID- 10816486 TI - Pilus-mediated adherence of Haemophilus influenzae to human respiratory mucins. AB - Haemophilus influenzae, especially the nontypeable strains, are among the most common pathogens encountered in patients with chronic lung disease and otitis media. We and others have demonstrated that respiratory isolates of nontypeable H. influenzae bind to human mucins, but the mechanism of binding is not entirely clear. We have therefore examined the role of pili in the adherence of both type b and nontypeable H. influenzae to human respiratory mucins. We used isogenic H. influenzae strains with a mutation in the structural gene for pilin (hifA), a laboratory H. influenzae strain transformed with a type b pilus gene cluster (from strain C54), antibodies raised against H. influenzae HifA, and Escherichia coli strains carrying a cloned type b pilus gene cluster (from strain AM30) in these studies. All bacteria lacking HifA or the pilus gene cluster had decreased adherence of piliated H. influenzae to mucins, and Fab fragments of anti-HifA antibodies inhibited the adherence. E. coli strains carrying the cloned type b pilus gene cluster were six to seven times more adhesive than strains carrying the vector. The role of other putative adhesins was not examined and thus cannot be excluded, but these studies support a role for pili in the binding of H. influenzae to human respiratory mucins. PMID- 10816487 TI - Transcriptional organization and function of invasion genes within Salmonella enterica serovar Typhimurium pathogenicity island 1, including the prgH, prgI, prgJ, prgK, orgA, orgB, and orgC genes. AB - Salmonella enterica serovar Typhimurium initiates infection of a host by inducing its own uptake into specialized M cells which reside within the epithelium overlaying Peyer's patches. Entry of Salmonella into intestinal epithelial cells is dependent upon invasion genes that are clustered together in Salmonella pathogenicity island 1 (SPI-1). Upon contact between serovar Typhimurium and epithelial cells targeted for bacterial internalization, bacterial proteins are injected into the host cell through a type III secretion system that leads to internalization of the bacteria. Previous work has established that the prgH, -I, -J, and -K and orgA genes reside in SPI-1, and the products of these genes are predicted to be components of the invasion secretion apparatus. We report that an error in the published orgA DNA sequence has been identified so that this region encodes two small genes rather than a single large open reading frame. These genes have been designated orgA and orgB. Additionally, an opening reading frame downstream of orgB, which we have designated orgC, has been identified and partially characterized. Previously published work has indicated that the prgH, I, -J, and -K genes are transcribed from a promoter distinct from that used by the gene immediately downstream, orgA. Here, we present experiments indicating that orgA expression is driven by the prgH promoter. In addition, using reverse transcriptase PCR analysis, we have found that this polycistronic message extends downstream of prgH to include a total of 10 genes. To more fully characterize this invasion operon, we demonstrate that the prgH, prgI, prgJ, prgK, orgA, and orgB genes are each required for invasion and secretion, while orgC is not essential for the invasive phenotype. PMID- 10816488 TI - Adhesion of Aspergillus species to extracellular matrix proteins: evidence for involvement of negatively charged carbohydrates on the conidial surface. AB - Invasive lung disease caused by Aspergillus species is a potentially fatal infection in immunocompromised patients. The adhesion of Aspergillus fumigatus conidia to proteins in the basal lamina is thought to be an initial step in the development of invasive aspergillosis. The purpose of this study was to determine the mechanism of adhesion of A. fumigatus conidiospores to basal-lamina proteins and to determine whether conidia possess unique adhesins which allow them to colonize the host. We compared conidia from different Aspergillus species for the ability to bind to purified fibronectin and intact basal lamina. Adhesion assays using immobilized fibronectin or type II pneumocyte-derived basal lamina showed that A. fumigatus conidia bound significantly better than those of other Aspergillus species to both fibronectin and intact basal lamina. Neither desialylation nor complete deglycosylation of fibronectin decreased the binding of A. fumigatus conidia to fibronectin, suggesting that oligosaccharides on fibronectin were not involved in conidiospore binding. Further evidence for this hypothesis came from experiments using purified fragments of fibronectin; A. fumigatus conidia preferentially bound to the nonglycosylated 40-kDa fragment which contains the glycosaminoglycan (GAG) binding domain. Negatively charged carbohydrates, including dextran sulfate and heparin, as well as high-ionic strength buffers, inhibited binding of A. fumigatus conidia to both fibronectin and intact basal lamina, suggesting that negatively charged carbohydrates on the surface of the conidium may bind to the GAG binding domain of fibronectin and other basal-lamina proteins. These data provide evidence for a novel mechanism of conidial attachment whereby adherence to fibronectin and other basal-lamina proteins is mediated via negatively charged carbohydrates on the conidial surface. PMID- 10816489 TI - Schistosoma mansoni phosphoenolpyruvate carboxykinase, a novel egg antigen: immunological properties of the recombinant protein and identification of a T cell epitope. AB - In schistosomiasis mansoni, hepatic granulomatous inflammation surrounding parasite eggs is mediated by CD4(+) T helper (Th) cells sensitized to schistosomal egg antigens (SEA). We previously showed that a prominent lymphoproliferative response of CD4(+) Th cells from schistosome-infected C57BL/6 (BL/6) mice was directed against a 62-kDa component of SEA. A partial amino acid sequence of the 62-kDa component was found to be identical with one present in the enzyme phosphoenolpyruvate carboxykinase (PEPCK). Based on this sequence, a cDNA clone containing the entire coding region of PEPCK was identified, and the full recombinant Schistosoma mansoni PEPCK (rSm-PEPCK) of 626 amino acids was purified from a prokaryotic expression system. rSm-PEPCK strongly stimulated a specific T-cell hybridoma, 4E6, as well as CD4(+) Th cells from SEA-immunized BL/6 mice and from infected BL/6, CBA, and BALB/c mice. In the infected mice, rSm PEPCK elicited significant gamma interferon production as well as, to a lesser extent, production of interleukin-2 and -5. In BL/6 and BALB/c mice, the CD4(+) Th cell response to rSm-PEPCK was greater than that directed against the egg antigen Sm-p40; conversely, CBA mice responded better to Sm-p40 than to Sm-PEPCK. A 12-amino-acid region (residues 398 to 409: DKSKDPKAHPNS) was demonstrated to contain a T-cell epitope; synthetic peptides containing this epitope significantly stimulated specific hybridoma 4E6 and polyclonal CD4(+) Th cells. The identification and characterization of immunogenic egg components will contribute to the understanding and possible control of T-cell-mediated schistosomal disease. PMID- 10816490 TI - Expression of interleukin-1beta, tumor necrosis factor alpha, and interleukin-6 in human peripheral blood leukocytes exposed to human granulocytic ehrlichiosis agent or recombinant major surface protein P44. AB - Human granulocytic ehrlichiosis (HGE) is an emerging febrile systemic disease caused by the HGE agent, an obligatory intracellular bacterium of granulocytes. The pathogenicity- and immunity-related mechanisms of HGE are unknown. In this study, several cytokines generated in human peripheral blood leukocytes (PBLs) incubated with the HGE agent or a recombinant 44-kDa major surface protein (rP44) of the HGE agent were examined by reverse transcription-PCR and a capture enzyme linked immunosorbent assay. The HGE agent induced expression of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and IL-6 mRNAs and proteins in PBLs in a dose-dependent manner to levels as high as those resulting from Escherichia coli lipopolysaccharide stimulation. The kinetics of induction of these three cytokines in PBLs by rP44 and by the HGE agent were similar. Proteinase K treatment of the HGE agent or rP44 eliminated the ability to induce these three cytokines. Induction of these cytokine mRNAs was not dependent on superoxide generation. These results suggest that P44 proteins have a major role in inducing the production of proinflammatory cytokines by PBLs. Expression of IL 8, IL-10, gamma interferon, transforming growth factor beta, and IL-2 mRNAs in response to the HGE agent was not remarkable. Among PBLs, neutrophils and lymphocytes expressed IL-1beta mRNA but not TNF-alpha or IL-6 mRNA in response to the HGE agent, whereas monocytes expressed all three of these cytokine mRNAs. These observations suggest that induction of proinflammatory-cytokine gene expression by the major outer membrane protein of the HGE agent in monocytes, which are not the primary host cells of the HGE agent, contributes to HGE pathogenesis and immunomodulation. PMID- 10816491 TI - A subdominant CD8(+) cytotoxic T lymphocyte (CTL) epitope from the Plasmodium yoelii circumsporozoite protein induces CTLs that eliminate infected hepatocytes from culture. AB - Previous studies indicated that the Plasmodium yoelii circumsporozoite protein (PyCSP) 57-70 region elicits T cells capable of eliminating infected hepatocytes in vitro. Herein, we report that the PyCSP58-67 sequence contains an H-2(d) binding motif, which binds purified K(d) molecules in vitro with low affinity (3, 267 nM) and encodes an H-2(d)-restricted cytotoxic T lymphocyte (CTL) epitope. Immunization of BALB/c mice with three doses of a multiple antigen peptide (MAP) construct containing four branches of amino acids 57 to 70 linked to a lysine glycine core [MAP4(PyCSP57-70)] and Lipofectin as the adjuvant induced both T cell proliferation and a peptide-specific CTL response that was PyCSP59-67 specific, H-2(d) restricted, and CD8(+) T cell dependent. Immunization with either DNA encoding the PyCSP or irradiated sporozoites demonstrated that this CTL epitope is subdominant since it is not recognized in the context of whole CSP immunization. The biological relevance of this CTL response was underlined by the demonstration that it could mediate genetically restricted, CD8(+)- and nitric oxide-dependent elimination of infected hepatocytes in vitro, as well as partial protection of BALB/c mice against sporozoite challenge. These findings indicate that subdominant epitopes with low major histocompatibility complex affinity can be used to engineer epitope-based vaccines and have implications for the selection of epitopes for subunit-based vaccines. PMID- 10816492 TI - Jejunal brush border microvillous alterations in Giardia muris-infected mice: role of T lymphocytes and interleukin-6. AB - Intestinal colonization with the protozoan Giardia causes diffuse brush border microvillous alterations and disaccharidase deficiencies, which in turn are responsible for intestinal malabsorption and maldigestion. The role of T cells and/or cytokines in the pathogenesis of Giardia-induced microvillous injury remains unclear. The aim of this study was to assess the role of T cells and interleukin-6 (IL-6) in the brush border pathophysiology of acute murine giardiasis in vivo. Athymic nude (nu(-)/nu(-)) CD-1 mice and isogenic immunocompetent (nu(+)/nu(+)) CD-1 mice (4 weeks old) received an axenic Giardia muris trophozoite inoculum or vehicle (control) via orogastric gavage. Weight gain and food intake were assessed daily. On day 6, segments of jejunum were assessed for parasite load, brush border ultrastructure, IL-6 content, maltase and sucrase activities, villus-crypt architecture, and intraepithelial lymphocyte (IEL) infiltration. Despite similar parasitic loads on day 6, infected immunocompetent animals, but not infected nude mice, showed a diffuse loss of brush border microvillous surface area, which was correlated with a significant reduction in maltase and sucrase activities and a decrease in jejunal IL-6 concentration. In both athymic control and infected mice, jejunal brush border surface area and disaccharidases were high, but levels of tissue IL-6 were low and comparable to the concentration measured in immunocompetent infected animals. In both immunocompetent and nude mice, infection caused a small but significant increase in the numbers of IELs. These findings suggest that the enterocyte brush border injury and malfunction seen in giardiasis is, at least in part, mediated by thymus-derived T lymphocytes and that suppressed jejunal IL-6 does not necessarily accompany microvillous shortening. PMID- 10816493 TI - The response regulator PhoP is important for survival under conditions of macrophage-induced stress and virulence in Yersinia pestis. AB - The two-component regulatory system PhoPQ has been identified in many bacterial species. However, the role of PhoPQ in regulating virulence gene expression in pathogenic bacteria has been characterized only in Salmonella species. We have identified, cloned, and sequenced PhoP orthologues from Yersinia pestis, Yersinia pseudotuberculosis, and Yersinia enterocolitica. To investigate the role of PhoP in the pathogenicity of Y. pestis, an isogenic phoP mutant was constructed by using a reverse-genetics PCR-based strategy. The protein profiles of the wild type and phoP mutant strains, grown at either 28 or 37 degrees C, revealed more than 20 differences, indicating that PhoP has pleiotrophic effects on gene expression in Y. pestis. The mutant showed a reduced ability to survive in J774 macrophage cell cultures and under conditions of low pH and oxidative stress in vitro. The mean lethal dose of the phoP mutant in mice was increased 75-fold in comparison with that of the wild-type strain, indicating that the PhoPQ system plays a key role in regulating the virulence of Y. pestis. PMID- 10816494 TI - Investigation into the mechanism of gamma interferon-mediated inhibition of Toxoplasma gondii in murine astrocytes. AB - Toxoplasma gondii is an obligate intracellular parasite that is a common opportunistic pathogen of the central nervous system in AIDS patients. Gamma interferon (IFN-gamma) alone or in combination with interleukin-1 (IL-1), IL-6, or tumor necrosis factor alpha significantly inhibits the growth of T. gondii in murine astrocytes, suggesting these are important nonimmune effector cells in the brain. Inhibition was found to be independent of a nitric oxide-mediated or tryptophan starvation mechanism. Both reactive oxygen intermediates and iron deprivation are IFN-gamma-mediated mechanisms known to operate against intracellular parasites in other cell types. Astrocytes generated from mice genetically deficient in the production of reactive oxygen intermediates (phox(-/ ) mice) were found to inhibit growth of T. gondii when stimulated with IFN-gamma alone or in combination with other cytokines. The reactive oxygen inhibitor catalase and the reactive oxygen scavengers mannitol and thiourea failed to reverse the IFN-gamma-induced inhibition of T. gondii in astrocytes. These data indicate that IFN-gamma-induced inhibition in astrocytes is independent of reactive oxygen intermediates. IFN-gamma-induced inhibition could not be reversed by the addition of iron salts, ferric citrate, ferric nitrate, or ferric transferrin. Pretreatment of astrocytes with desferrioxamine also did not induce the inhibition of T. gondii. These data indicate that the mechanism of IFN-gamma inhibition was not due to iron deprivation. IFN-gamma had no effect on T. gondii invasion of astrocytes, but inhibition of growth and loss of tachyzoite vacuoles were evident in IFN-gamma-treated astrocytes by 24 h after invasion. Overall, these data suggest that IFN-gamma-activated astrocytes inhibit T. gondii by an as yet-unknown mechanism. PMID- 10816495 TI - Afa/Dr diffusely adhering Escherichia coli C1845 infection promotes selective injuries in the junctional domain of polarized human intestinal Caco-2/TC7 cells. AB - The Afa/Dr diffusely adhering Escherichia coli (DAEC) C1845 strain harboring the F1845 fimbrial adhesin interacts with the brush border-associated CD55 molecule and promotes elongation of brush border microvilli resulting from rearrangement of the F-actin network. This phenomenon involves the activation of a cascade of signaling coupled to the glycosylphosphatidylinositol-anchored receptor of the F1845 adhesin. We provide evidence that infection of the polarized human intestinal cell line Caco-2/TC7 by strain C1845 is followed by an increase in the paracellular permeability for [(3)H]mannitol without a decrease of the transepithelial resistance of the monolayers. Alterations in the distribution of tight-junction (TJ)-associated occludin and ZO-1 protein are observed, whereas the distribution of the zonula adherens-associated E-cadherin is not affected. Using the recombinant E. coli strains HB101(pSSS1) and -(pSSS1C) expressing the F1845 fimbrial adhesin, we demonstrate that the adhesin-CD55 interaction is not sufficient for the induction of structural and functional TJ lesions. Moreover, using the actin filament-stabilizing agent Jasplakinolide, we demonstrate that the C1845-induced functional alterations in TJs are independent of the C1845 induced apical cytoskeleton rearrangements. The results indicated that pathogenic factor(s) other than F1845 adhesin may be operant in Afa/Dr DAEC C1845. PMID- 10816496 TI - Identification of a human lactoferrin-binding protein in Gardnerella vaginalis. AB - Previous studies have shown that Gardnerella vaginalis can utilize iron-loaded human lactoferrin as a sole source of iron. In this study, G. vaginalis cells were shown to bind digoxigenin (DIG)-labeled human lactoferrin in a dot blot assay. Using the DIG-labeled human lactoferrin, a 120-kDa human lactoferrin binding protein was detected by Western blot analysis of G. vaginalis proteins. The lactoferrin-binding activity of this protein was found to be heat stable. Competition studies indicated that this binding activity was specific for human lactoferrin. Treatment of G. vaginalis cells with proteases suggested that this protein was surface exposed. An increase in lactoferrin binding by the 120-kDa protein was observed in G. vaginalis cells grown under iron-restrictive conditions, suggesting that this activity may be iron regulated. PMID- 10816497 TI - Immune response against a cross-reactive epitope on the heat shock protein 60 homologue of Helicobacter pylori. AB - We previously established a monoclonal antibody (MAb), designated H9, which reacts with the heat shock protein 60 (HSP60) homologue of Helicobacter pylori as well as with other bacterial and human HSP60s. To determine the importance of a cross-reactive epitope on H. pylori HSP60 in H. pylori immunopathogenesis, we performed (i) mapping of an epitope on H. pylori HSP60 recognized by the H9 MAb, (ii) analysis of immunoglobulin G responses of patients with or without H. pylori infection to its epitope region, and (iii) studies of the protective effect of immunization with its epitope region on H. pylori infection in mice. The epitope recognized by the H9 MAb was mapped to the sequence of amino acids 189 to 203 (VEGMQFDRGYLSPYF) on the H. pylori HSP60 molecule. It was confirmed that the synthesized peptide designated pH9 was recognized by the H9 MAb. Enzyme-linked immunosorbent assay analysis showed that patients with H. pylori infection (n = 349) had significantly lower titers of pH9 antibody than did uninfected patients (n = 200) (P < 0.001), but this was not the case with purified H. pylori HSP60 recombinant Escherichia coli GroEL, or recombinant human HSP60. In C57BL/6 mice immunized with the pH9 peptide with Freund's complete adjuvant (FCA), the number of H. pylori organisms colonizing the stomach was significantly lower than that in mice immunized with pCont plus FCA (P < 0.0001) or FCA only (P < 0.005). The results suggest that the immune response to the cross-reactive epitope (pH9 region) on H. pylori HSP60 is unique and might be associated with protection against H. pylori infection. PMID- 10816498 TI - A model for sequestration of the transmission stages of Plasmodium falciparum: adhesion of gametocyte-infected erythrocytes to human bone marrow cells. AB - With the aim of developing an appropriate in vitro model of the sequestration of developing Plasmodium falciparum sexual-stage parasites, we have investigated the cytoadherence of gametocytes to human bone marrow cells of stromal and endothelial origin. Developing stage III and IV gametocytes, but not mature stage V gametocytes, adhere to bone marrow cells in significantly higher densities than do asexual-stage parasites, although these adhesion densities are severalfold lower than those encountered in classical CD36-dependent assays of P. falciparum cytoadherence. This implies that developing gametocytes undergo a transition from high-avidity, CD36-mediated adhesion during stages I and II to a lower-avidity adhesion during stages III and IV. We show that this adhesion is CD36 independent, fixation sensitive, stimulated by tumor necrosis factor alpha, and dependent on divalent cations and serum components. These data suggest that gametocytes and asexual parasites utilize distinct sets of receptors for adhesion during development in their respective sequestered niches. To identify receptors for gametocyte-specific adhesion of infected erythrocytes to bone marrow cells, we tested a large panel of antibodies for the ability to inhibit cytoadherence. Our results implicate ICAM-1, CD49c, CD166, and CD164 as candidate bone marrow cell receptors for gametocyte adhesion. PMID- 10816499 TI - The capsule is a virulence determinant in the pathogenesis of Pasteurella multocida M1404 (B:2). AB - Capsules from a range of pathogenic bacteria are key virulence determinants, and the capsule has been implicated in virulence in Pasteurella multocida. We have previously identified and determined the nucleotide sequence of the P. multocida M1404 (B:2) capsule biosynthetic locus (J. D. Boyce, J. Y. Chung, and B. Adler, Vet. Microbiol. 72:121-134, 2000). The cap locus consists of 15 genes, which can be grouped into three functional regions. Regions 1 and 3 contain genes proposed to encode proteins involved in capsule export, and region 2 contains genes proposed to encode proteins involved in polysaccharide biosynthesis. In order to construct a mutant impaired in capsule export, the final gene of region 1, cexA, was disrupted by insertion of a tetracycline resistance cassette by allelic replacement. The genotype of the tet(M) OmegacexA mutant was confirmed by Southern hybridization and PCR. The acapsular phenotype was confirmed by immunofluorescence, and the strain could be complemented and returned to capsule production by the presence of a cloned uninterrupted copy of cexA. Wild-type, mutant, and complemented strains were tested for virulence by intraperitoneal challenge of mice; the presence of the capsule was shown to be a crucial virulence determinant. Following intraperitoneal challenge of mice, the acapsular bacteria were removed efficiently from the blood, spleen, and liver, while wild type bacteria multiplied rapidly. Acapsular bacteria were readily taken up by murine peritoneal macrophages, but wild-type bacteria were significantly resistant to phagocytosis. Both wild-type and acapsular bacteria were resistant to complement in bovine and murine serum. PMID- 10816500 TI - Characterization of hemolysin of Moraxella bovis using a hemolysis-neutralizing monoclonal antibody. AB - A concentrated bacterial culture supernatant from the hemolytic Moraxella bovis strain UQV 148NF was used to immunize mice and generate monoclonal antibodies (MAbs). One, MAb G3/D7, neutralized the hemolytic activity of M. bovis and recognized a 94-kDa protein by Western blot analysis in hemolytic M. bovis strains representing each of the different fimbrial serogroups. Exposure of corneal epithelial cells to M. bovis concentrated culture supernatants demonstrated a role for an exotoxin in the pathogenesis of infectious bovine keratoconjunctivitis, while neutralization of hemolytic and cytotoxic activities by MAb G3/D7 implies that these activities are related or have common epitopes. The action of M. bovis hemolysin was further characterized in sheep erythrocyte preparations with a binding step and Ca(2+) required for lysis to proceed, similar to the RTX family of bacterial exotoxins. Neutralization of lytic activity in vitro is evidence for the presence of M. bovis antigens, which may be capable of protecting cattle from the development of infectious bovine keratoconjunctivitis. PMID- 10816501 TI - Clostridium perfringens iota toxin: binding studies and characterization of cell surface receptor by fluorescence-activated cytometry. AB - The binding characteristics of iota toxin, a binary enterotoxin produced by Clostridium perfringens type E, were studied by fluorescence-activated cytometry. The proteolytically activated binding component of iota toxin, iota b (Ib), bound to various cell types when incubated at 4, 25, or 37 degrees C for 10 min. The binding of Ib was inhibited by antisera against C. perfringens type E or Clostridium spiroforme culture supernatants, but not C. perfringens types C or D. Pretreatment of Vero cells with glycosidases or lectins did not affect Ib interactions, while pronase effectively prevented Ib binding to the cell surface. The Ib protomer (Ibp) bound to the cell surface, but trypsinization of Ibp was necessary for docking of the ADP-ribosylating component, iota a (Ia). Ia attached to cell-bound Ib within 10 min at 37 degrees C, but surface levels of Ia decreased 90% after 30 min and were undetectable by 60 min. Detectable surface levels of Ib also diminished over time, and Western blot analysis suggested internalization or embedment of Ib into the membrane. PMID- 10816502 TI - Role of hyphal formation in interactions of Candida albicans with endothelial cells. AB - The ability to change from yeast to hyphal morphology is a major virulence determinant of Candida albicans. Mutants with defined defects in filamentation regulatory pathways have reduced virulence in mice. However, is it poorly understood why hyphal formation is critical for C. albicans to cause hematogenously disseminated infections. We used recently constructed mutants to examine the role of hyphal formation in the interactions of C. albicans with endothelial cells in vitro. These interactions included the ability of the mutants to invade and injure endothelial cells. Because the formation of hyphae may influence the host inflammatory response to C. albicans, we also investigated the capacity of these mutants to stimulate endothelial cells to express E selectin and intercellular adhesion molecule 1. We infected endothelial cells with C. albicans strains containing homozygous null mutations in the following filamentation regulatory genes: CLA4, CPH1, EFG1, and TUP1. Whereas the wild-type strain formed true hyphae on endothelial cells, we found that neither the Deltaefg1 nor the Deltacph1 Deltaefg1 double mutant germinated. The Deltatup1 mutant formed only pseudohyphae. We also found that the Deltaefg1, Deltacph1 Deltaefg1, and Deltatup1 mutants had significantly reduced capacities to invade and injure endothelial cells. Therefore, Efg1p and Tup1p contribute to virulence by regulating hyphal formation and the factors that enable C. albicans to invade and injure endothelial cells. With the exception of the Deltacph1 Deltaefg1 mutant, all other mutants stimulated endothelial cells to express at least one of the leukocyte adhesion molecules. Therefore, the combined activities of Cph1p and Efg1p are required for C. albicans to stimulate a proinflammatory response in endothelial cells. PMID- 10816504 TI - Role of interleukin-18 (IL-18) during lethal shock: decreased lipopolysaccharide sensitivity but normal superantigen reaction in IL-18-deficient mice. AB - Lethal shock can be associated with excessive secretion of cytokines such as tumor necrosis factor (TNF) and gamma interferon (IFN-gamma). IFN-gamma mediates macrophage activation and appears to be controlled by interleukin (IL)-12 and IL 18. To investigate the role of IL-18 in vivo, we generated IL-18-deficient mice by gene targeting. IL-18(-/-) mice showed decreased sensitivity towards lipopolysaccharide (LPS)-induced shock. LPS-induced IFN-gamma production was abrogated, yet induction of IL-12 and TNF was not affected. Both wild-type and IL 18-deficient mice succumbed to LPS-induced lethal shock after sensitization with D-galactosamine. However, in marked contrast to LPS, the bacterial superantigen Staphylococcus aureus enterotoxin B (SEB) induced comparable serum levels of IFN gamma in IL-18(+/+) and IL-18(-/-) mice, accompanied by an upregulation of cell surface markers CD14, CD122 (IL-2Rbeta), and CD132 (IL-2Rgamma) on peritoneal macrophages. Moreover, SEB injection rendered IL-18-deficient mice sensitive for subsequent challenge with LPS. The degree of sensitization was comparable to that in wild-type controls with respect to lethality. However, LPS-induced TNF levels in serum were significantly reduced in SEB-sensitized IL-18-deficient mice. These results imply that IL-18 plays an important role in induction of IFN-gamma and lethality in response to LPS. PMID- 10816503 TI - Identification of potential vaccine and drug target candidates by expressed sequence tag analysis and immunoscreening of Onchocerca volvulus larval cDNA libraries. AB - The search for appropriate vaccine candidates and drug targets against onchocerciasis has so far been confronted with several limitations due to the unavailability of biological material, appropriate molecular resources, and knowledge of the parasite biology. To identify targets for vaccine or chemotherapy development we have undertaken two approaches. First, cDNA expression libraries were constructed from life cycle stages that are critical for establishment of Onchocerca volvulus infection, the third-stage larvae (L3) and the molting L3. A gene discovery effort was then initiated by random expressed sequence tag analysis of 5,506 cDNA clones. Cluster analyses showed that many of the transcripts were up-regulated and/or stage specific in either one or both of the cDNA libraries when compared to the microfilariae, L2, and both adult stages of the parasite. Homology searches against the GenBank database facilitated the identification of several genes of interest, such as proteinases, proteinase inhibitors, antioxidant or detoxification enzymes, and neurotransmitter receptors, as well as structural and housekeeping genes. Other O. volvulus genes showed homology only to predicted genes from the free-living nematode Caenorhabditis elegans or were entirely novel. Some of the novel proteins contain potential secretory leaders. Secondly, by immunoscreening the molting L3 cDNA library with a pool of human sera from putatively immune individuals, we identified six novel immunogenic proteins that otherwise would not have been identified as potential vaccinogens using the gene discovery effort. This study lays a solid foundation for a better understanding of the biology of O. volvulus as well as for the identification of novel targets for filaricidal agents and/or vaccines against onchocerciasis based on immunological and rational hypothesis-driven research. PMID- 10816505 TI - Inhibition of adhesion of Escherichia coli k88ac fimbria to its receptor, intestinal mucin-type glycoproteins, by a monoclonal antibody directed against a variable domain of the fimbria. AB - Strains of enterotoxigenic Escherichia coli that express K88 fimbriae are among the most common causes of diarrhea in young pigs. Adhesion of bacteria to receptors on intestinal epithelial cells, mediated by K88 fimbriae, is the initial step in the establishment of infection. Three antigenic variants of K88 fimbriae exist in nature: K88ab, K88ac, and K88ad. K88ac is the most prevalent and may be the only variant of significance in swine disease. Each K88 fimbrial variant is composed of multiple antigenic determinants. Some of these determinants are shared among the three variants and may be referred to as conserved epitopes, whereas others are unique to a specific variant and may be referred to as variable epitopes. In this study, monoclonal antibodies (MAbs) specific to either variable or conserved epitopes of K88ac fimbriae were produced. The specificity of each MAb was tested by enzyme-linked immunosorbent and immunoblot assays. Fab fragments were prepared from these MAbs and were tested for their ability to block the binding of K88-positive bacteria and purified fimbriae to porcine enterocyte brush border vesicles and purified K88 receptors, respectively. The purified receptors were intestinal mucin-type sialoglycoproteins (IMTGP) isolated from porcine enterocytes (A. K. Erickson, D. R. Baker, B. T. Bosworth, T. A. Casey, D. A. Benfield, and D. H. Francis, Infect. Immun. 62:5404-5410, 1994). Fab fragments prepared from MAbs specific for variable epitopes blocked the binding of bacteria to brush borders and of fimbriae to IMTGP. However, those from MAbs specific for a conserved epitope did not. These observations indicate that the receptor-binding domain of a K88ac fimbria is contained, at least in part, within the antigenically variable epitopes of that fimbria. Epitope mapping for one of the MAbs, which recognizes a linear epitope on K88ac fimbriae, indicated that this MAb binds to the region from amino acid no. 64 to no. 107 on the major subunit of K88ac fimbriae. PMID- 10816506 TI - Expression of Staphylococcus aureus clumping factor A in Lactococcus lactis subsp. cremoris using a new shuttle vector. AB - Staphylococcus aureus harbors redundant adhesins mediating tissue colonization and infection. To evaluate their intrinsic role outside of the staphylococcal background, a system was designed to express them in Lactococcus lactis subsp. cremoris 1363. This bacterium is devoid of virulence factors and has a known genetic background. A new Escherichia coli-L. lactis shuttle and expression vector was constructed for this purpose. First, the high-copy-number lactococcal plasmid pIL253 was equipped with the oriColE1 origin, generating pOri253 that could replicate in E. coli. Second, the lactococcal promoters P23 or P59 were inserted at one end of the pOri253 multicloning site. Gene expression was assessed by a luciferase reporter system. The plasmid carrying P23 (named pOri23) expressed luciferase constitutively at a level 10,000 times greater than did the P59-containing plasmid. Transcription was absent in E. coli. The staphylococcal clumping factor A (clfA) gene was cloned into pOri23 and used as a model system. Lactococci carrying pOri23-clfA produced an unaltered and functional 130-kDa ClfA protein attached to their cell walls. This was indicated both by the presence of the protein in Western blots of solubilized cell walls and by the ability of ClfA positive lactococci to clump in the presence of plasma. ClfA-positive lactococci had clumping titers (titer of 4,112) similar to those of S. aureus Newman in soluble fibrinogen and bound equally well to solid-phase fibrinogen. These experiments provide a new way to study individual staphylococcal pathogenic factors and might complement both classical knockout mutagenesis and modern in vivo expression technology and signature tag mutagenesis. PMID- 10816507 TI - Genetic relatedness and superantigen expression in group A streptococcus serotype M1 isolates from patients with severe and nonsevere invasive diseases. AB - The relatedness of group A streptococcal (GAS) strains isolated from 35 Canadian patients with invasive disease of different severity was investigated by a variety of molecular methods. All patients were infected with M1T1 strains and, based on clinical criteria, were classified as severe (n = 21) and nonsevere (n = 14) invasive GAS infection cases. All the M1 strains studied had the emm1.0 allele and the same streptococcal pyrogenic exotoxin (Spe) genotype, speA(+) speB(+) speC speF(+) speG(+) speH smeZ(+) ssa. All isolates had the same speA allotype, speA2. The randomly amplified polymorphic DNA banding pattern with two different primers was identical for all strains, and pulsed field gel electrophoresis analysis showed that 33 and 30 isolates had identical banding patterns after DNA digestion with SfiI or SmaI, respectively; the nonidentical isolates differed from the main pattern by only one band. A relatively high degree of polymorphism in specific regions of the sic gene was observed among isolates; however, this polymorphism was not associated with disease severity. Likewise, although the phenotypic expression of SpeA, SpeB, and SpeF proteins varied among the M1T1 isolates, there was no correlation between the amount of Spe expressed and disease severity. Importantly, mitogenic and cytokine responses induced by partially purified bacterial culture supernatants containing a mixture of expressed superantigens were very similar for isolates from severe and nonsevere cases (P > 0.1). Together, the data indicate that highly related invasive M1T1 isolates, some indistinguishable, can cause disease of varying severity in different individuals. These findings underscore the contribution of host factors to the outcome of invasive GAS infections. PMID- 10816508 TI - Bacterial cell wall-induced arthritis: chemical composition and tissue distribution of four Lactobacillus strains. AB - To study what determines the arthritogenicity of bacterial cell walls, cell wall induced arthritis in the rat was applied, using four strains of Lactobacillus. Three of the strains used proved to induce chronic arthritis in the rat; all were Lactobacillus casei. The cell wall of Lactobacillus fermentum did not induce chronic arthritis. All arthritogenic bacterial cell walls had the same peptidoglycan structure, whereas that of L. fermentum was different. Likewise, all arthritogenic cell walls were resistant to lysozyme degradation, whereas the L. fermentum cell wall was lysozyme sensitive. Muramic acid was observed in the liver, spleen, and lymph nodes in considerably larger amounts after injection of an arthritogenic L. casei cell wall than following injection of a nonarthritogenic L. fermentum cell wall. The L. casei cell wall also persisted in the tissues longer than the L. fermentum cell wall. The present results, taken together with those published previously, underline the possibility that the chemical structure of peptidoglycan is important in determining the arthritogenicity of the bacterial cell wall. PMID- 10816509 TI - Receptor structure for F1C fimbriae of uropathogenic Escherichia coli. AB - F1C fimbriae are correlated with uropathogenic Escherichia coli strains. Although F1C fimbriae mediate binding to kidney tubular cells, their receptor is not known. In this paper, we demonstrate for the first time specific carbohydrate residues as receptor structure for F1C-fimbria-expressing E. coli. The binding of the F1C fimbriated recombinant E. coli strain HB101(pPIL110-54) and purified F1C fimbriae to reference glycolipids of different carbohydrate compositions was evaluated by using thin-layer chromatography (TLC) overlay and solid-phase binding assays. TLC fimbrial overlay analysis revealed the binding ability of purified F1C fimbriae only to glucosylceramide (GlcCer), beta1-linked galactosylceramide 2 (GalCer2) with nonhydroxy fatty acids, lactosylceramide, globotriaosylceramide, paragloboside (nLc(4)Cer), lactotriaosylceramide, gangliotriaosylceramide (asialo-GM(2) [GgO(3)Cer]) and gangliotetraosylceramide (asialo-GM(1) [GgO(4)Cer]). The binding of purified F1C fimbriae as well as F1C fimbriated recombinant E. coli strain HB101(pPIL110-54) was optimal to microtiter plates coated with asialo-GM(2) (GgO(3)Cer). The bacterial interaction with asialo-GM(1) (GgO(4)Cer) and asialo-GM(2) (GgO(3)Cer) was strongly inhibited only by disaccharide GalNAcbeta1-4Galbeta linked to bovine serum albumin. We observed no binding to globotetraosylceramide or Forssman antigen (Gb(5)Cer) glycosphingolipids or to sialic-acid-containing gangliosides. It was demonstrated that the presence of a GalCer or GlcCer residue alone is not sufficient for optimal binding, and additional carbohydrate residues are required for high affinity adherence. Indeed, the binding efficiency of F1C fimbriated recombinant bacteria increased by 19-fold when disaccharide sequence GalNAcbeta1-4Galbeta is linked to glucosylceramide as in asialo-GM(2) (GgO(3)Cer). Thus, it is suggested that the disaccharide sequence GalNAcbeta1-4Galbeta of asialo-GM(2) (GgO(3)Cer) which is positioned internally in asialo-GM(1) (GgO(4)Cer) is the high-affinity binding epitope for the F1C fimbriae of uropathogenic E. coli. PMID- 10816510 TI - In vitro resistance of Staphylococcus aureus to thrombin-induced platelet microbicidal protein is associated with alterations in cytoplasmic membrane fluidity. AB - Platelet microbicidal proteins (PMPs) are small, cationic peptides which possess potent microbicidal activities against common bloodstream pathogens, such as Staphylococcus aureus. We previously showed that S. aureus strains exhibiting resistance to thrombin-induced PMP (tPMP-1) in vitro have an enhanced capacity to cause human and experimental endocarditis (T. Wu, M. R. Yeaman, and A. S. Bayer, Antimicrob. Agents Chemother. 38:729-732, 1994; A. S. Bayer et al., Antimicrob. Agents Chemother. 42:3169-3172, 1998; V. K. Dhawan et al., Infect. Immun. 65:3293 3299, 1997). However, the mechanisms mediating tPMP-1 resistance in S. aureus are not fully delineated. The S. aureus cell membrane appears to be a principal target for the action of tPMP-1. To gain insight into the basis of tPMP-1 resistance, we compared several parameters of membrane structure and function in three tPMP-1-resistant (tPMP-1(r)) strains and their genetically related, tPMP-1 susceptible (tPMP-1(s)) counterpart strains. The tPMP-1(r) strains were derived by three distinct methods: transposon mutagenesis, serial passage in the presence of tPMP-1 in vitro, or carriage of a naturally occurring multiresistance plasmid (pSK1). All tPMP-1(r) strains were found to possess elevated levels of longer chain, unsaturated membrane lipids, in comparison to their tPMP-1(s) counterparts. This was reflected in corresponding differences in cell membrane fluidity in the strain pairs, with tPMP-1(r) strains exhibiting significantly higher degrees of fluidity as assessed by fluorescence polarization. These data provide further support for the concept that specific alterations in the cytoplasmic membrane of S. aureus strains are associated with tPMP-1 resistance in vitro. PMID- 10816511 TI - Recruitment of CD55 and CD66e brush border-associated glycosylphosphatidylinositol-anchored proteins by members of the Afa/Dr diffusely adhering family of Escherichia coli that infect the human polarized intestinal Caco-2/TC7 cells. AB - The Afa/Dr family of diffusely adhering Escherichia coli (Afa/Dr DAEC) includes bacteria expressing afimbrial adhesins (AFA), Dr hemagglutinin, and fimbrial F1845 adhesin. We show that infection of human intestinal Caco-2/TC7 cells by the Afa/Dr DAEC strains C1845 and IH11128 is followed by clustering of CD55 around adhering bacteria. Mapping of CD55 epitopes involved in CD55 clustering by Afa/Dr DAEC was conducted using CD55 deletion mutants expressed by stable transfection in CHO cells. Deletion in the short consensus repeat 1 (SCR1) domain abolished Afa/Dr DAEC-induced CD55 clustering. In contrast, deletion in the SCR4 domain does not modify Afa/Dr DAEC-induced CD55 clustering. We show that the brush border-associated glycosylphosphatidylinositol (GPI)-anchored protein CD66e (carcinoembryonic antigen) is recruited by the Afa/Dr DAEC strains C1845 and IH11128. This conclusion is based on the observations that (i) infection of Caco 2/TC7 cells by Afa/Dr DAEC strains is followed by clustering of CD66e around adhering bacteria and (ii) Afa/Dr DAEC strains bound efficiently to stably transfected HeLa cells expressing CD66e, accompanied by CD66e clustering around adhering bacteria. Inhibition assay using monoclonal antibodies directed against CD55 SCR domains, and polyclonal anti-CD55 and anti-CD66e antibodies demonstrate that CD55 and CD66e function as a receptors for the C1845 and IH11128 bacteria. Moreover, using structural draE gene mutants, we found that a mutant in which cysteine replaced aspartic acid at position 54 displayed conserved binding capacity but failed to induce CD55 and CD66e clustering. Taken together, these data give new insights into the mechanisms by which Afa/Dr DAEC induces adhesin dependent cross talk in the human polarized intestinal epithelial cells by mobilizing brush border-associated GPI-anchored proteins known to function as transducing molecules. PMID- 10816512 TI - A recombinant baculovirus-expressed Plasmodium falciparum receptor-binding domain of erythrocyte binding protein EBA-175 biologically mimics native protein. AB - EBA-175 of Plasmodium falciparum is a merozoite ligand that binds its receptor glycophorin A on erythrocytes during invasion. The ligand-receptor interaction is dependent on sialic acids as well as the protein backbone of glycophorin A. Region II (RII) of EBA-175 has been defined as the receptor-binding domain. RII is divided into regions F1 and F2, which contain duplicated cysteine motifs. We expressed RII in a baculovirus and show that RII binds erythrocytes with a specificity identical to that of the native protein. We found that, consistent with the binding of erythrocytes to COS cells expressing F2, recombinant baculovirus-expressed F2 bound erythrocytes. About 20% of all baculovirus expressed RII is N-glycosylated, unlike native P. falciparum proteins that remain essentially unglycosylated. However, glycosylation of recombinant RII did not affect its immunogenicity. Antibodies raised against both glycosylated and unglycosylated baculovirus-expressed RII recognized P. falciparum schizonts in immunofluorescence assays and also gave similar enzyme-linked immunosorbent assay titers. Furthermore, these antibodies have similar abilities to block native EBA 175 binding to erythrocytes. These results allow the development of RII as a vaccine candidate for preclinical assessment. PMID- 10816513 TI - Metalloprotease is not essential for Vibrio vulnificus virulence in mice. AB - Previous work suggested that a metalloprotease, Vvp, may be a virulence factor of Vibrio vulnificus, which causes severe wound infection and septicemia in humans. To determine the role of Vvp in pathogenesis, we isolated an isogenic protease deficient (PD) mutant of Vibrio vulnificus by in vivo allelic exchange. This PD mutant was as virulent as its parental strain in mice infected intraperitoneally and was 10-fold more virulent in mice infected via the oral route. Furthermore, the PD mutant was indistinguishable from its parental strain in invasion from peritoneal cavity into blood stream, enhancement of vascular permeability, growth in murine blood, and utilization of hemoglobin and transferrin. These data suggest that Vvp is not essential for virulence in the mouse. However, the cytolysin activity in the culture supernatant of the PD mutant was found to be twofold higher than that of the wild-type strain and remained for a much longer period. The higher cytolysin activity of the PD mutant may be associated with the enhanced virulence in mice infected via the oral route. PMID- 10816514 TI - The SA85-1.1 protein of the Trypanosoma cruzi trans-sialidase superfamily is a dominant T-cell antigen. AB - Trypanosoma cruzi currently infects 18 million people, and 30% of those infected develop a chronic inflammatory process that causes significant morbidity or mortality. The major histocompatibility complex class II (MHC-II)-restricted T cell response is critical to the control of the infection and to the ensuing inflammatory pathology. The specific epitopes or major antigens of this response have not been identified. The parasite simultaneously expresses variant members of the trans-sialidase superfamily. To begin to analyze the MHC-II response to these variant proteins, the response to a single surface protein, SA85-1.1, was initiated. These studies have demonstrated that a biased gamma interferon (IFN gamma) response to the SA85-1.1 protein develops during T. cruzi infection. In addition, adoptive transfer of a CD4 clone that recognizes an SA85-1.1 epitope, named epitope 1, and immunization with a peptide encoding epitope 1 were protective and suggested that epitope 1 may be immunodominant. In this report IFN gamma intracellular staining demonstrated that splenocytes from acutely and chronically infected mice, incubated with SA85-1.1 protein or peptides that encode epitope 1, result in IFN-gamma synthesis by 4 to 6% of the splenic CD4 cells. These data indicate that during T. cruzi infection epitope 1 is a major epitope and that 4 to 6% of the CD4 cells are stimulated by a single trans sialidase superfamily epitope and suggest that a combination of trans-sialidase superfamily proteins combines to stimulate a majority of CD4 cells. These data suggest that during T. cruzi infection the CD4 response to the trans-sialidase superfamily is critical to the protective response and to the ensuing chronic inflammatory pathology. PMID- 10816515 TI - Gram-positive bacteria are potent inducers of monocytic interleukin-12 (IL-12) while gram-negative bacteria preferentially stimulate IL-10 production. AB - Interleukin-10 (IL-10) and IL-12 are two cytokines secreted by monocytes/macrophages in response to bacterial products which have largely opposite effects on the immune system. IL-12 activates cytotoxicity and gamma interferon (IFN-gamma) secretion by T cells and NK cells, whereas IL-10 inhibits these functions. In the present study, the capacities of gram-positive and gram negative bacteria to induce IL-10 and IL-12 were compared. Monocytes from blood donors were stimulated with UV-killed bacteria from each of seven gram-positive and seven gram-negative bacterial species representing both aerobic and anaerobic commensals and pathogens. Gram-positive bacteria induced much more IL-12 than did gram-negative bacteria (median, 3,500 versus 120 pg/ml at an optimal dose of 25 bacteria/cell; P < 0.001), whereas gram-negative bacteria preferentially stimulated secretion of IL-10 (650 versus 200 pg/ml; P < 0.001). Gram-positive species also induced stronger major histocompatibility complex class II restricted IFN-gamma production in unfractionated blood mononuclear cells than did gram-negative species (12,000 versus 3,600 pg/ml; P < 0.001). The poor IL-12 inducing capacity of gram-negative bacteria was not remediated by addition of blocking anti-IL-10 antibodies to the cultures. No isolated bacterial component could be identified that mimicked the potent induction of IL-12 by whole gram positive bacteria, whereas purified LPS induced IL-10. The results suggest that gram-positive bacteria induce a cytokine pattern that promotes Th1 effector functions. PMID- 10816516 TI - Cooperation between reactive oxygen and nitrogen intermediates in killing of Rhodococcus equi by activated macrophages. AB - Rhodococcus equi is a facultative intracellular bacterium of macrophages which can infect immunocompromised humans and young horses. In the present study, we examine the mechanism of host defense against R. equi by using a murine model. We show that bacterial killing is dependent upon the presence of gamma interferon (IFN-gamma), which activates macrophages to produce reactive nitrogen and oxygen intermediates. These two radicals combine to form peroxynitrite (ONOO(-)), which kills R. equi. Mice deficient in the production of either the high-output nitric oxide pathway (iNOS(-/-)) or the oxidative burst (gp91(phox-/-)) are more susceptible to lethal R. equi infection and display higher bacterial burdens in their livers, spleens, and lungs than wild-type mice. These in vivo observations, which implicate both nitric oxide (NO) and superoxide (O(2)(-)) in bacterial killing, were reexamined in cell-free radical-generating assays. In these assays, R. equi remains fully viable following prolonged exposure to high concentrations of either nitric oxide or superoxide, indicating that neither compound is sufficient to mediate bacterial killing. In contrast, brief exposure of bacteria to ONOO(-) efficiently kills virulent R. equi. The intracellular killing of bacteria in vitro by activated macrophages correlated with the production of ONOO(-) in situ. Inhibition of nitric oxide production by activated macrophages by using N(G)-monomethyl-L-arginine blocks their production of ONOO(-) and weakens their ability to control rhodococcal replication. These studies indicate that peroxynitrite mediates the intracellular killing of R. equi by IFN-gamma activated macrophages. PMID- 10816517 TI - Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct. AB - Strains of Staphylococcus aureus were transformed with plasmid DNA containing a Photorhabdus luminescens lux operon (luxABCDE) that was genetically modified to be functional in both gram-positive and gram-negative bacteria. S. aureus cells containing this novel lux construct, downstream of an appropriate promoter sequence, are highly bioluminescent, allowing the detection of fewer than 100 CFU in vitro (direct detection of exponentially dividing cells in liquid culture). Furthermore, these bacteria produce light stably at 37 degrees C and do not require exogenous aldehyde substrate, thus allowing S. aureus infections in living animals to be monitored by bioluminescence. Two strains of S. aureus 8325 4 that produce high levels of constitutive bioluminescence were injected into the thigh muscles of mice, and the animals were then either treated with the antibiotic amoxicillin or left untreated. Bioluminescence from bacteria present in the thighs of the mice was monitored in vivo over a period of 24 h. The effectiveness of the antibiotic in the treated animals could be measured by a decrease in the light signal. At 8 h, the infection in both groups of treated animals had begun to clear, as judged by a decrease in bioluminescence, and by 24 h no light signal could be detected. In contrast, both groups of untreated mice had strong bioluminescent signals at 24 h. Quantification of CFU from bacteria extracted from the thigh muscles of the mice correlated well with the bioluminescence data. This paper shows for the first time that bioluminescence offers a method for monitoring S. aureus infections in vivo that is sensitive and noninvasive and requires fewer animals than conventional methodologies. PMID- 10816518 TI - Carcinoembryonic antigen family receptor specificity of Neisseria meningitidis Opa variants influences adherence to and invasion of proinflammatory cytokine activated endothelial cells. AB - The carcinoembryonic antigen (CEA) family member CEACAM1 (previously called biliary glycoprotein or CD66a) was previously shown to function as a receptor that can mediate the binding of Opa protein-expressing Neisseria meningitidis to both neutrophils and epithelial cells. Since neutrophils and polarized epithelia have both been shown to coexpress multiple CEACAM receptors, we have now extended this work to characterize the binding specificity of meningococcal Opa proteins with other CEA family members. To do so, we used recombinant Escherichia coli expressing nine different Opa variants from three meningococcal strains and stably transfected cell lines expressing single members of the CEACAM family. These infection studies demonstrated that seven of the nine Opa variants bound to at least one CEACAM receptor and that binding to each of these receptors is sufficient to trigger the Opa-dependent bacterial uptake by these cell lines. The other two Opa variants do not appear to bind to either CEACAM receptors or heparan sulfate proteoglycan receptors, which are bound by some gonococcal Opa variants, thus implying a novel class of Opa proteins. We have also extended previous studies by demonstrating induction of CEACAM1 expression after stimulation of human umbilical vein endothelial cells with the proinflammatory cytokine tumor necrosis factor alpha, which is present in high concentrations during meningococcal disease. This induced expression of CEACAM1 leads to an increased Opa-dependent bacterial binding and invasion into the primary endothelia, implying that these interactions may play an important role in the pathogenesis of invasive meningococcal disease. PMID- 10816519 TI - Shigella flexneri IpaH(7.8) facilitates escape of virulent bacteria from the endocytic vacuoles of mouse and human macrophages. AB - The behavior of Shigella flexneri ipaH mutants was studied in human monocyte derived macrophages (HMDM), in 1-day-old human monocytes, and in J774 mouse macrophage cell line. In HMDM, strain pWR700, an ipaH(7.8) deletion mutant of S. flexneri 2a strain 2457T, behaved like the wild-type strain 2457T. This strain caused rapid host cell death by oncosis, and few bacterial CFU were recovered after incubation in the presence of gentamicin as previously described for 2457T infected HMDM. However, analysis of bacterial compartmentalization within endocytic vacuoles with gentamicin and chloroquine indicated that more pWR700 than 2457T was present within the endocytic vacuoles of HMDM, suggesting that ipaH(7.8) deletion mutant transited more slowly from the vacuoles to the cytoplasm. In contrast to findings with HMDM, CFU recovered from pWR700-infected mouse J774 cells were 2 to 3 logs higher than CFU from 2457T-infected J774 cells. These values exceeded CFU recovered after infection of J774 cells with plasmid cured avirulent strain M4243A1. Incubation with gentamicin and chloroquine clearly showed that pWR700 within J774 cells was mostly present within the endocytic vacuoles. This distribution pattern was similar to that seen with M4243A1 and contrasted with the pattern seen with 2457T. Complementation of pWR700 with a recombinant clone expressing ipaH(7. 8) restored the intracellular distribution of bacteria to that seen with the wild-type strain. Strains with deletions in ipaH(4.5) or ipaH(9.8), however, behaved like 2457T in both HMDM and J774 cells. The distribution profile of pWR700 in 1-day-old monocytes was similar to that seen in J774 cells. Like infected J774 cells, 1-day-old human monocytes demonstrated apoptosis upon infection with virulent Shigella. These results suggest that a role of the ipaH(7. 8) gene product is to facilitate the escape of the virulent bacteria from the phagocytic vacuole of monocytes and macrophages. PMID- 10816520 TI - Innate immune responses in children and adults with Shigellosis. AB - An array of pro- and anti-inflammatory mediators of the innate immune system was analyzed in stool, urine, and rectal mucosa samples from adults and children with shigellosis to better understand their role in recovery from and in the immunopathogenesis of the disease. Increased concentrations of lactoferrin (Lf), myeloperoxidase (MPO), prostaglandin E(2), and leukotriene B(4) (LTB(4)) in stool during acute shigellosis in both children and adults indicated that activated cells of the innate defense system at the mucosal site were secreting the mediators. Increased concentration of MPO and 8-iso-prostaglandin F(2alpha) and lower levels of superoxide dismutase (SOD) activity in stool during acute Shigella infection suggested increased formation of reactive oxygen species, free radical-catalyzed peroxidation of membrane lipids, and decreased scavenging of the reactive oxygen radicals. In children, lower expression of SOD in tissue with severe inflammation and lower levels of SOD activity in stool for longer periods compared to adults may further worsen the tissue damage and predispose the children to a lowered defense. Both adult and pediatric patients had significantly higher expression of inducible nitric oxide synthase (iNOS) in the rectum with severe inflammation, compared to that seen with mild inflammation, accompanied by persistently up-regulated iNOS mRNA, reflecting increased production of nitric oxide at the local site. However, in contrast to adults, reduced urinary nitrate levels in pediatric patients during acute shigellosis suggested lower production of nitric oxide in the renal compartment. Persistent production of Lf in pediatric patients may contribute to chronic inflammation in the rectum. In addition, increased production of proinflammatory mediators in the rectum of patients with severe histology suggested contribution of these molecules to the immunopathogenesis of severe colitis caused by shigellae. PMID- 10816521 TI - Pyrogenic toxin superantigen site specificity in toxic shock syndrome and food poisoning in animals. AB - Staphylococcus aureus and Streptococcus pyogenes express pyrogenic toxin superantigens (PTSAgs) that are associated with toxic shock syndrome (TSS) and staphylococcal food poisoning (SFP). Most PTSAgs cause TSS in deep-tissue infections, whereas only TSS toxin 1 (TSST-1) is associated with menstrual, vaginal TSS. In contrast, SFP has been linked only with staphylococcal enterotoxins (SEs). Because of the differential abilities of PTSAgs to cause systemic or localized symptoms in a site-dependent manner, the present study was undertaken to assess the toxins' abilities to cross mucosal barriers. The activity of three PTSAgs when delivered orally, vaginally, or intravenously to rabbits and orally to monkeys was investigated. TSST-1 induced shock via all three routes in rabbits. Although active when administered intravenously, SEC1 and streptococcal pyrogenic exotoxin A (SPEA) did not cause symptoms when administered orally or vaginally. Only SEC1 induced emesis in the monkey feeding assay. TSST-1, albeit less stable than SEC1 and SPEA to pepsin, induced diarrhea in monkeys. Our results may explain the unique association of TSST-1 with menstrual TSS and why SPEA is only rarely associated with TSS after pharyngitis, despite being highly associated with TSS after subcutaneous infections. Finally, our studies indicate that enterotoxicity in SFP is not the result of superantigenicity. PMID- 10816522 TI - Production of basic fibroblast growth factor and interleukin 6 by human smooth muscle cells following infection with Chlamydia pneumoniae. AB - Chlamydia pneumoniae infection has been associated with asthma and atherosclerosis. Smooth muscle cells represent host cells for chlamydiae during chronic infection. In this study we demonstrated that C. pneumoniae infection of human smooth muscle cells in vitro increased production of interleukin 6 (IL-6) and basic fibroblast growth factor (bFGF) as shown by reverse transcription-PCR, immunoblotting, and enzyme-linked immunosorbent assay. In contrast, levels of platelet-derived growth factor A-chain mRNA were not affected after infection. The stimulation of bFGF and IL-6 production was most effective when viable chlamydiae were used as inoculum. Furthermore, inhibition of bacterial protein synthesis with chloramphenicol prevented up-regulation of IL-6 and bFGF in infected cells. Addition of IL-6 antibody to infected cultures diminished bFGF expression, indicating involvement of produced IL-6. These findings suggest that chlamydial infection of smooth muscle cells elicits a cytokine response that may contribute to structural remodeling of the airway wall in chronic asthma and to fibrous plaque formation in atherosclerosis. PMID- 10816523 TI - Antibody interactions with the capsule of Cryptococcus neoformans. AB - Monoclonal antibodies to the encapsulated fungus Cryptococcus neoformans produce different immunofluorescence (IF) patterns after binding to the polysaccharide capsule. To explore the relationship between the IF pattern and the location of antibody binding, two immunoglobulin M (IgM) monoclonal antibodies (MAbs) (12A1 and 13F1) that differ in protective efficacy and IF pattern and one protective IgG1 MAb (2H1) were studied by IF and electron microscopy (EM). Fixing C. neoformans cells in lung tissue for EM resulted in significantly better preservation of the capsule than fixing yeast cells in suspension. The localization of MAbs 12A1 and 13F1 by immunogold EM differed depending on whether the MAb was bound to cells in cut tissue sections embedded in plastic or to cells in solution. In cut tissue sections, MAbs 12A1 and 13F1 bound throughout the capsule, whereas in solution both MAbs bound near the capsule surface. To investigate whether antibody binding to the C. neoformans capsule affected the binding of other primary or secondary reagents, various combinations of MAbs 12A1, 13F1, and 2H1 were studied by direct and indirect IF. The IF pattern and location of binding for MAbs 12A1, 13F1, and 2H1 varied depending on the presence of other capsule-binding MAbs and the method of detection. The results show that (i) binding of MAbs to the C. neoformans polysaccharide capsule can modify the binding of subsequent primary or secondary antibodies; (ii) the IgM MAbs bind primarily to the outer capsule regions despite the occurrence of their epitopes throughout the capsule; and (iii) MAb 2H1 staining of newly formed buds is reduced, suggesting quantitative or qualitative differences in bud capsule. PMID- 10816524 TI - Drastic reduction of a filarial infection in eosinophilic interleukin-5 transgenic mice. AB - In order to establish the role of eosinophils in destroying parasites, transgenic mice have been used in experimental helminthiases but not in filariasis. Litomosoides sigmodontis offers a good opportunity for this study because it is the only filarial species that completes its life cycle in mice. Its development was compared in transgenic CBA/Ca mice overexpressing interleukin-5 (IL-5) and in wild-type mice following subcutaneous inoculation of 40 infective larvae. An acceleration of larval growth was observed in the IL-5 transgenic mice. However, the recovery rate of adult worms was considerably reduced in these mice, as evidenced 2 months postinoculation (p.i.). The reduction occurs between days 10 and 30 p.i. in the coelomic cavities. As early as day 10, spherical aggregates of eosinophils and macrophages are seen attached on live developing larvae (always similarly localized on the worm) in both wild-type and transgenic mice. However, on day 60 p.i., granulomas were found in the transgenic mice only, probably because of the higher density of eosinophils. Furthermore, on day 30 p.i., young filariae are seen trapped in granulomas, some of them surrounded by Splendore Hoeppli deposits, which illustrates the release of the major basic protein by eosinophils. The high protection rate obtained (65%) is similar to that observed previously in BALB/c mice following vaccination with irradiated larvae. Both protocols have a common factor, the high production of IL-5 and eosinophilia. However, protection occurs later in primary infected transgenic mice because specific antibodies are not yet present at the time of challenge. PMID- 10816525 TI - An OmpA-like protein from Acinetobacter spp. stimulates gastrin and interleukin-8 promoters. AB - Bacterial overgrowth in the stomach may occur under conditions of diminished or absent acid secretion. Under these conditions, secretion of the hormone gastrin is elevated. Alternatively, bacterial factors may directly stimulate gastrin. Consistent with this hypothesis, we found that mice colonized for 2 months with a mixed bacterial culture of opportunistic pathogens showed an increase in serum gastrin. To examine regulation of gene expression by bacterial proteins, stable transformants of AGS cells expressing gastrin or interleukin-8 (IL-8) promoters were cocultured with live organisms. Both whole-cell sonicates and a heat-stable fraction were also coincubated with the cells. A level of 10(8) organisms per ml stimulated both the gastrin and IL-8 promoters. Heat-stable proteins prepared from these bacterial sonicates stimulated the promoter significantly more than the live organism or unheated sonicates. A 38-kDa heat-stable protein stimulating the gastrin and IL-8 promoters was cloned and found to be an OmpA-related protein. Immunoblotting using antibody to the OmpA-like protein identified an Acinetobacter sp. as the bacterial species that expressed this protein and colonized the mouse stomach. Moreover, reintubation of mice with a pure culture of the Acinetobacter sp. caused gastritis. We conclude that bacterial colonization of the stomach may increase serum gastrin levels in part through the ability of the bacteria to produce OmpA-like proteins that directly stimulate gastrin and IL-8 gene expression. These results implicate OmpA-secreting bacteria in the activation of gastrin gene expression and raise the possibility that a variety of organisms may contribute to the increase in serum gastrin and subsequent epithelial cell proliferation in the hypochlorhydric stomach. PMID- 10816526 TI - Antibodies against thrombospondin-related anonymous protein do not inhibit Plasmodium sporozoite infectivity in vivo. AB - Thrombospondin-related anonymous protein (TRAP), a candidate malaria vaccine antigen, is required for Plasmodium sporozoite gliding motility and cell invasion. For the first time, the ability of antibodies against TRAP to inhibit sporozoite infectivity in vivo is evaluated in detail. TRAP contains an A-domain, a well-characterized adhesive motif found in integrins. We modeled here a three dimensional structure of the TRAP A-domain of Plasmodium yoelii and located regions surrounding the MIDAS (metal ion-dependent adhesion site), the presumed business end of the domain. Mice were immunized with constructs containing these A-domain regions but were not protected from sporozoite challenge. Furthermore, monoclonal and rabbit polyclonal antibodies against the A-domain, the conserved N terminus, and the repeat region of TRAP had no effect on the gliding motility or sporozoite infectivity to mice. TRAP is located in micronemes, secretory organelles of apicomplexan parasites. Accordingly, the antibodies tested here stained cytoplasmic TRAP brightly by immunofluorescence. However, very little TRAP could be detected on the surface of sporozoites. In contrast, a dramatic relocalization of TRAP onto the parasite surface occurred when sporozoites were treated with calcium ionophore. This likely mimics the release of TRAP from micronemes when a sporozoite contacts its target cell in vivo. Contact with hepatoma cells in culture also appeared to induce the release of TRAP onto the surface of sporozoites. If large amounts of TRAP are released in close proximity to its cellular receptor(s), effective competitive inhibition by antibodies may be difficult to achieve. PMID- 10816527 TI - Lack of protection in mice and necrotizing bronchointerstitial pneumonia with bronchiolitis in guinea pigs immunized with vaccines directed against the hsp60 molecule of Mycobacterium tuberculosis. AB - In this study, the hsp60 and hsp70 heat shock protein antigens of Mycobacterium tuberculosis were tested as potential vaccine candidates, using purified recombinant protein antigens or antigens encoded in the form of a DNA plasmid vaccine. Guinea pigs vaccinated with a mixture of the two proteins showed no evidence of resistance to low-dose aerosol challenge infection and quickly developed severe lung damage characterized by necrotizing bronchointerstitial pneumonia and bronchiolitis. As a result, we turned instead to a DNA vaccination approach using a plasmid encoding the hsp60 antigen of M. tuberculosis. Although immunogenic in mice, vaccination with plasmid DNA encoding hsp60 was not protective in that model or in the guinea pig model and again gave rise to similar severe lung damage. This study seriously questions the safety of vaccines against tuberculosis that target highly conserved heat shock proteins. PMID- 10816528 TI - Listeria monocytogenes-infected human dendritic cells: uptake and host cell response. AB - Dendritic cells (DCs) are potent antigen-presenting cells and play a crucial role in initiation and modulation of specific immune responses. Various pathogens are able to persist inside DCs. However, internalization of the gram-positive bacterium Listeria monocytogenes into human DCs has not yet been shown. In the present study, we demonstrate that human monocyte-derived immature DCs can efficiently phagocytose L. monocytogenes. This uptake is independent of listerial adhesion factors internalin A and internalin B but requires cytoskeletal motion and factors present in human plasma. A major portion of internalized bacteria is found in membrane-bound phagosomes and is rarely free in the cytosol, as shown by transmission electron microscopy and by using an L. monocytogenes strain expressing green fluorescent protein when in the host cell cytosol. The infection caused maturation of the immature DCs into mature DCs displaying high levels of CD83, CD25, major histocompatibility complex class II, and the CD86 costimulator molecule. This effect appeared to be largely mediated by listerial lipoteichoic acid. Although L. monocytogenes infection is known to induce death in other cell types, infection of human DCs was found to induce necrotic but not apoptotic death in fewer than 20% of DCs. Therefore, the ability of DCs to act as effective antigen-presenting cells for listerial immunity is probably enhanced by their resistance to cell death, as well as their ability to rapidly differentiate into mature, immunostimulatory DCs upon encountering bacteria. PMID- 10816529 TI - Role of EspB in experimental human enteropathogenic Escherichia coli infection. AB - Enteropathogenic Escherichia coli (EPEC), a leading cause of diarrhea among infants in developing countries, induces dramatic alterations in host cell architecture that depend on a type III secretion system. EspB, one of the proteins secreted and translocated to the host cytoplasm via this system, is required for numerous alterations in host cell structure and function. To determine the role of EspB in virulence, we conducted a randomized, double-blind trial comparing the ability of wild-type EPEC and an isogenic DeltaespB mutant strain to cause diarrhea in adult volunteers. Diarrhea developed in 9 of 10 volunteers who ingested the wild-type strain but in only 1 of 10 volunteers who ingested the DeltaespB mutant strain. Marked destruction of the microvillous brush border adjacent to adherent organisms was observed in a jejunal biopsy from a volunteer who ingested the wild-type strain but not from two volunteers who ingested the DeltaespB mutant strain. Humoral and cell-mediated immune responses to EPEC antigens were stronger among recipients of the wild-type strain. In addition, four of the volunteers who ingested the wild-type strain had lymphoproliferative responses to EspB. These results demonstrate that EspB is a critical virulence determinant of EPEC infections and suggest that EspB contributes to an immune response. PMID- 10816530 TI - Biosynthesis and functions of melanin in Sporothrix schenckii. AB - Sporothrix schenckii is a human pathogen that causes sporotrichosis, an important cutaneous mycosis with a worldwide distribution. It produces dark-brown conidia, which infect the host. We found that S. schenckii synthesizes melanin via the 1,8 dihydroxynaphthalene pentaketide pathway. Melanin biosynthesis in the wild type was inhibited by tricyclazole, and colonies of the fungus were reddish brown instead of black on tricyclazole-amended medium. Two melanin-deficient mutant strains were analyzed in this study: an albino that produced normal-appearing melanin on scytalone-amended medium and a reddish brown mutant that accumulated and extruded melanin metabolites into its medium. Scytalone and flaviolin obtained from cultures of the reddish brown mutant were identified by thin-layer chromatography, high-performance liquid chromatography, and UV spectra. Transmission electron microscopy showed an electron-dense granular material believed to be melanin in wild-type conidial cell walls, and this was absent in conidial walls of the albino mutant unless the albino was grown on a scytalone amended medium. Melanized cells of wild-type S. schenckii and the albino grown on scytalone-amended medium were less susceptible to killing by chemically generated oxygen- and nitrogen-derived radicals and by UV light than were conidia of the mutant strains. Melanized conidia of the wild type and the scytalone-treated albino were also more resistant to phagocytosis and killing by human monocytes and murine macrophages than were unmelanized conidia of the two mutants. These results demonstrate that melanin protects S. schenckii against certain oxidative antimicrobial compounds and against attack by macrophages. PMID- 10816531 TI - In vivo administration of mycobacterial cord factor (Trehalose 6, 6'-dimycolate) can induce lung and liver granulomas and thymic atrophy in rabbits. AB - Trehalose 6,6'-dimycolate (TDM) is a cell surface molecule of Mycobacterium tuberculosis. TDM induced a loss of body weight and prominent granulomas in the liver and lungs by the intravenous injection of TDM into rabbits. TDM also induced atrophy of the thymus and spleen due to apoptosis. By contrast, sulfolipid (2,3,6, 6'-tetraacyl trehalose 2'-sulfate) induced neither toxicity, nor granuloma formation, nor atrophy of the thymus and spleen. In rabbits the histopathological changes were more dramatic than in mice. The rabbit model may be more sensitive and may provide more information on the beneficial or pathological effects of TDM. PMID- 10816532 TI - Interaction of Shigella flexneri IpaC with model membranes correlates with effects on cultured cells. AB - Invasion of enterocytes by Shigella flexneri requires the properly timed release of IpaB and IpaC at the host-pathogen interface; however, only IpaC has been found to possess quantifiable activities in vitro. We demonstrate here that when added to cultured cells, purified IpaC elicits cytoskeletal changes similar to those that occur during Shigella invasion. This IpaC effect may correlate with its ability to interact with model membranes at physiological pH and to promote entry by an ipaC mutant of S. flexneri. PMID- 10816533 TI - Replacement of histidine 340 with alanine inactivates the group A Streptococcus extracellular cysteine protease virulence factor. AB - Streptococcus pyogenes expresses a highly conserved extracellular cysteine protease that is a virulence factor for invasive disease, including soft tissue infection. Site-directed mutagenesis was used to generate a His340Ala recombinant mutant protein that was made as a stable 40-kDa zymogen by Escherichia coli. Purified His340Ala protein was proteolytically inactive when bovine casein and human fibronectin were used as substrates. Wild-type 28-kDa streptococcal protease purified from S. pyogenes processed the 40-kDa mutant zymogen to a 28 kDa mature form, a result suggesting that the derivative protein retained structural integrity. The data are consistent with the hypothesis that His340 is an enzyme active site residue, an idea confirmed by recent solution of the zymogen crystal structure (T. F. Kagawa, J. C. Cooney, H. M. Baker, S. McSweeney, M. Liu, S. Gubba, J. M. Musser, and E. N. Baker, Proc. Natl. Acad. Sci. USA 97:2235-2240, 2000). The data provide additional insight into structure-function relationships in this S. pyogenes virulence factor. PMID- 10816534 TI - Gamma interferon is essential for clearing Mycobacterium genavense infection. AB - Factors determining the in vivo replication of the opportunistic pathogen Mycobacterium genavense are largely unknown. Following intravenous injection of a patient isolate, M. genavense could not be recovered by culture or detected by PCR in the livers or spleens of infected BALB/c mice. In contrast, M. genavense was found to chronically persist and multiply in the livers and spleens of intravenously infected syngeneic gamma-interferon-gene-deficient (GKO) mice as evidenced by acid-fast stains of infected tissues and recovery by both PCR and liquid culture following organ homogenization. In GKO mice, M. genavense elicited a chronic inflammatory response, resulting in marked splenomegaly and extensive lymphadenopathy. Granulomatous lesions in the livers of GKO mice were diffuse, were composed of monocytes, neutrophils, and CD3(+) cells, and were histochemically negative for inducible nitric oxide synthase. PMID- 10816535 TI - Do kinetics of the humoral response to Cryptococcus neoformans proteins during murine cryptococcosis reflect outcome? AB - The kinetics of the humoral response to Cryptococcus neoformans proteins were studied in outbred mice infected with isolate NIH52D. Future nonsurvivors had earlier and stronger (i.e., more bands recognized) humoral responses than survivors. In addition, antibodies to a 56- to 60-kDa membrane antigen and to a 39- to 40-kDa cytosolic antigen were detected more frequently in samples from future nonsurvivors and from survivors, respectively (P < 0.05). PMID- 10816536 TI - Stimulation of Bordetella pertussis adenylate cyclase toxin intoxication by its hemolysin domain. AB - The internalization of the N-terminal catalytic domain of Bordetella pertussis adenylate cyclase toxin (ACT) across the cytoplasmic membrane has been considered to occur independently from protein-protein interactions which can lead to oligomerization required for hemolytic activity by its C-terminal hemolysin domain. Here we report that when added in excess, this hemolysin domain stimulates the internalization, suggesting the involvement of protein-protein interactions in cell-invasive activity of ACT, as well as its hemolytic activity. PMID- 10816537 TI - Toll-like receptors confer responsiveness to lipopolysaccharide from Porphyromonas gingivalis in human gingival fibroblasts. AB - Gingival fibroblasts produce proinflammatory cytokines in response to lipopolysaccharide (LPS) from periodontopathic bacteria. Recently it has become evident that the human homologue of Drosophila Toll can transduce intracellular signaling by LPS stimulation. Toll-like receptors (TLRs) have been identified in myeloid cells; however, their role in nonmyeloid cells such as gingival fibroblasts has not been fully elucidated. Here, we report that human gingival fibroblasts constitutively express TLR2 and TLR4 and that their levels of expression are increased by stimulation with LPS from Porphyromonas gingivalis. Upregulated expression of interleukin-6 gene and protein in fibroblasts stimulated with LPS is inhibited by anti-TLR4 antibody. These findings suggest that TLRs may confer responsiveness to LPS in gingival fibroblasts. PMID- 10816538 TI - Interaction of human phagocytes with pigmentless Aspergillus conidia. AB - A defect in the pksP gene of Aspergillus fumigatus is associated with the loss of conidial pigmentation, a profound change of the conidial surface structure, and reduced virulence. The structural change of the conidial surface structure was not observed in similar A. nidulans wA mutants. Our data indicate that the pigment of both species is important for scavenging reactive oxygen species and for protection of conidia against oxidative damage. PMID- 10816539 TI - Detection of 23 immunogenic pneumococcal proteins using convalescent-phase serum. AB - A genomic expression library of Streptococcus pneumoniae was screened with a convalescent-phase serum for immunoreactive proteins. Six known and 17 unknown pneumococcal proteins were detected. Five of the known proteins were surface located virulence factors, and eight of the unknown proteins were putative membrane proteins. PMID- 10816540 TI - Salmonella enterica serovars Typhimurium and Dublin can lyse macrophages by a mechanism distinct from apoptosis. AB - Salmonella enterica serovars Typhimurium and Dublin lysed primary bovine alveolar macrophages and immortalized J774.2 macrophage-like cells in the absence of either the morphological changes or DNA fragmentation characteristic of apoptosis. Macrophage lysis was dependent on a subset of caspases and an intact sipB gene. PMID- 10816541 TI - Administration of endotoxin associated with lipopolysaccharide tolerance protects mice against fungal infection. AB - Lipopolysaccharide (LPS) pretreatment of mice resulted in a significantly enhanced survival after disseminated Cryptococcus neoformans infection. The survival was associated with reduced fungal burden in tissues. LPS-pretreated mice had lower levels of cytokines in blood, spleen, and lungs and higher levels in brain. Pentoxifylline abolished the beneficial effect of LPS pretreatment. PMID- 10816542 TI - Cell specificity of Helicobacter pylori cytotoxin is determined by a short region in the polymorphic midregion. AB - There are two alleles of the vacuolating cytotoxin gene from Helicobacter pylori, which code for toxins with different cell specificities. By analyzing the phenotypes of natural and artificial chimeras between the two forms of the protein, we have delimited a short stretch of amino acids which determine the cell specificity. PMID- 10816543 TI - Constitutive mutations of the Salmonella enterica serovar Typhimurium transcriptional virulence regulator phoP. AB - The PhoP-PhoQ two-component system is necessary for the virulence of Salmonella spp. and is responsible for regulating several modifications of the lipopolysaccharide (LPS). Mutagenesis of the transcriptional regulator phoP resulted in the identification of a mutant able to activate transcription of regulated genes approximately 100-fold in the absence of PhoQ. Sequence analysis showed two single-base alterations resulting in amino acid changes at positions 93 (S93N) and 203 (Q203R). These mutations were individually created, and although each resulted in a constitutive phenotype, the double mutant displayed a synergistic effect both in the induction of PhoP-activated gene expression and in resistance to antimicrobial peptides. The constitutive phoP gene was placed under the control of an arabinose-inducible promoter to examine the kinetics of PhoP activated gene induction and the resultant modifications of LPS. Gene induction and 2-hydroxymyristate modification of the lipid A were shown to occur within minutes of the addition of arabinose and to peak at 4 h. As the first constitutive mutant of phoP identified, this allele will be invaluable to future genetic and biochemical studies of this and likely other regulatory systems. PMID- 10816544 TI - Use of pertussis toxin encoded by ptx genes from Bordetella bronchiseptica to model the effects of antigenic drift of pertussis toxin on antibody neutralization. AB - Recently, concern has been voiced about the potential effect that antigenic divergence of circulating strains of Bordetella pertussis might have on the efficacy of pertussis vaccines. In order to model antigenic drift of pertussis toxin, a critical component of many pertussis vaccines, and to examine the effects of such drift on antibody neutralization, we engineered a strain of B. pertussis to produce a variant pertussis toxin molecule that contains many of the amino acid changes found in the toxin encoded by Bordetella bronchiseptica ptx genes. This altered form of the toxin, which is efficiently secreted by B. pertussis and which displays significant biological activity, was found to be neutralized by antibodies induced by vaccination as readily as toxin produced by wild-type B. pertussis. These findings suggest that significant amino acid changes in the pertussis toxin sequence can occur without drastically altering the ability of antibodies to recognize and neutralize the toxin molecule. PMID- 10816545 TI - Mutation of waaN reduces Salmonella enterica serovar Typhimurium-induced enteritis and net secretion of type III secretion system 1-dependent proteins. AB - Mutation of waaN, a gene involved in lipid A biosynthesis, reduced enteropathogenic responses induced by Salmonella enterica serovar Typhimurium in bovine ligated ileal loops. However, the secretion of key virulence determinants was also reduced, and therefore the reduction in enteropathogenicity cannot be solely attributed to a reduction in biological activity of lipid A. PMID- 10816546 TI - A functional cra gene is required for Salmonella enterica serovar typhimurium virulence in BALB/c mice. AB - A minitransposon mutant of Salmonella enterica serovar Typhimurium SR-11, SR-11 Fad(-), is unable to utilize gluconeogenic substrates as carbon sources and is avirulent and immunogenic when administered perorally to BALB/c mice (M. J. Utley et al., FEMS Microbiol. Lett., 163:129-134, 1998). Here, evidence is presented that the mutation in SR-11 Fad(-) that renders the strain avirulent is in the cra gene, which encodes the Cra protein, a regulator of central carbon metabolism. PMID- 10816547 TI - The collagen-binding adhesin is a virulence factor in Staphylococcus aureus keratitis. AB - A collagen-binding strain of Staphylococcus aureus produced suppurative inflammation in a rabbit model of soft contact lens-associated bacterial keratitis more often than its collagen-binding-negative isogenic mutant. Reintroduction of the cna gene on a multicopy plasmid into the mutant helped it regain its corneal adherence and infectivity. The topical application of a collagen-binding peptide before bacterial challenge decreased S. aureus adherence to deepithelialized corneas. These data suggest that the collagen-binding adhesin is involved in the pathogenesis of S. aureus infection of the cornea. PMID- 10816548 TI - Curli loci of Shigella spp. AB - An unstable chromosomal element encoding multiple antibiotic resistance in Shigella flexneri serotype 2a was found to include sequences homologous to the csg genes encoding curli in Escherichia coli and Salmonella enterica serovar Typhimurium. As curli have been implicated in the virulence of serovar Typhimurium, we investigated the csg loci in all four species of Shigella. DNA sequencing and PCR analysis showed that the csg loci of a wide range of Shigella strains, of diverse serotypes and different geographical distributions, were almost universally disrupted by deletions or insertions, indicating the existence of a strong selective pressure against the expression of curli. Strains of enteroinvasive E. coli (EIEC), which share virulence traits with Shigella spp. and cause similar diseases in humans, also possessed insertions or deletions in the csg locus or were otherwise unable to produce curli. Since the production of curli is a widespread trait in environmental isolates of E. coli, our results suggest that genetic lesions that abolish curli production in the closely related genus Shigella and in EIEC are pathoadaptive mutations. PMID- 10816549 TI - Reduced transendothelial migration of monocytes infected by Coxiella burnetii. AB - The migratory properties of THP1 monocytes infected by Coxiella burnetii were determined in a transmigration assay across a human microvascular endothelial cell monolayer. Transendothelial migration of monocytes infected by virulent, but not avirulent, C. burnetii was inhibited. This inhibition was observed in spite of conserved adherence properties of infected monocytes. PMID- 10816550 TI - Characterization of an anticryptococcal protein isolated from human serum. AB - Human serum at low concentrations inhibits the growth of Cryptococcus neoformans in vitro. Fractionation of serum yielded a purified inhibitory protein with a molecular mass of approximately 81.8 kDa, a pI of approximately 6.2, and an amino acid sequence that matched that of human transferrin. The inhibitory activity and that of apotransferrin and 5% human serum were reversed by 10 microM freshly prepared FeCl(3). PMID- 10816551 TI - Overexpression of a nuclear protein, TIEG, mimics transforming growth factor-beta action in human osteoblast cells. AB - Although transforming growth factor-beta (TGF-beta) is a growth factor with many known regulatory activities in many different cell types, its intracellular signaling pathway is still not fully understood. A TGF-beta-inducible early gene (TIEG) was discovered and shown by this laboratory to be a 3-zinc finger transcription factor family member; its expression is rapidly induced in cells treated with TGF-beta. To ascertain whether TIEG plays a major role in the TGF beta pathway, human osteosarcoma MG-63 cells were stably transfected either with an expression vector containing a TIEG cDNA or with the vector alone. Clones that contain only the vector express normal levels of TIEG mRNA and protein and display the same patterns of gene expression and levels of cell proliferation as the nontransfected, non-TGF-beta-treated parental cells. However, transfected cells that overexpress TIEG mRNA and protein (TIEG-6 and TIEG-7) display changes that mimic those of MG-63 cells treated with TGF-beta, i.e. increased alkaline phosphatase activity, decreased levels of osteocalcin mRNA and protein, and decreased cell proliferation. The degree of these changes correlated with the level of TIEG expressed in the cell lines. TGF-beta treatment of the overexpressed cells showed no added effects. These findings and other published reports support a primary role of TIEG as a transcription factor in the TGF-beta signaling pathway. PMID- 10816552 TI - A natural dominant negative P2X1 receptor due to deletion of a single amino acid residue. AB - The P2X1 receptor belongs to a family of oligomeric ATP-gated ion channels with intracellular N and C termini and two transmembrane segments separating a large extracellular domain. Here, we describe a naturally occurring dominant negative P2X1 mutant. This mutant lacks one leucine within a stretch of four leucine residues in its second transmembrane domain (TM2) (amino acids 351-354). Confocal microscopy revealed proper plasma membrane localization of the mutant in stably transfected HEK293 cells. Nevertheless, voltage-clamped HEK293 cells expressing mutated P2X1 channels failed to develop an ATP or ADP-induced current. Furthermore, when co-expressed with the wild type receptor in Xenopus oocytes, the mutated protein exhibited a dose-dependent dominant negative effect on the normal ATP or ADP-induced P2X1 channel activity. These data indicate that deletion of a single apolar amino acid residue at the inner border of the P2X1 TM2 generates a nonfunctional channel. The inactive and dominant negative form of the P2X1 receptor may constitute a new tool for the study of the physiological role of this channel in native cells. PMID- 10816553 TI - Light-induced down-regulation of the rat class 1 dynein-associated protein robl/LC7-like gene in visual cortex. AB - Dynein and kinesin are the main microtubule-dependent motors that mediate intracellular movement in eukaryotic organisms. We have cloned a full-length cDNA encoding rat dynein light chain protein, robl/LC7-like (class 1), from visual cortex. We found that rat robl/LC7-like gene is highly expressed in neocortex and displays the unusual feature of being rapidly down-regulated by sensory stimulation. This effect was seen at both mRNA and protein levels in visual cortex, being detectable in as little as 45 min after the onset of visual stimulation. Down-regulation by sensory stimulation was also found within ocular dominance columns of area V1 in monocularly deprived monkeys. Our results suggest a high turnover rate of the robl/LC7-like protein and the presence of a repressor mechanism in neurons that is tightly coupled to synaptic stimulation. PMID- 10816554 TI - Human alpha 1,3/4 fucosyltransferases. Characterization of highly conserved cysteine residues and N-linked glycosylation sites. AB - Human alpha1,3 fucosyltransferases (FucTs) contain four highly conserved cysteine (Cys) residues, in addition to a free Cys residue that lies near the binding site for GDP-fucose (Holmes, E. H., Xu, Z. , Sherwood, A. L., and Macher, B. A. (1995) J. Biol. Chem. 270, 8145-8151). The participation of the highly conserved Cys residues in disulfide bonds and their functional significance were characterized by mass spectrometry (MS) analyses and site-directed mutagenesis, respectively. Among the human FucTs is a subset of enzymes (FucT III, V, and VI) having highly homologous sequences, especially in the catalytic domain, and Cys residues in FucT III and V were characterized. The amino acid sequence of FucT III was characterized. Peptides containing the four conserved Cys residues were detected after reduction and alkylation, and found to be involved in disulfide bonds. The disulfide bond pattern was characterized by multiple stage MS analysis and the use of Glu-C protease and MS/MS analysis. Disulfide bonds in FucT III occur between Cys residues (Cys(81) to Cys(338) and Cys(91) to Cys(341)) at the N and C termini of the catalytic domain, bringing these ends close together in space. Mutagenesis of highly conserved Cys residues to Ser in FucT V resulted in proteins lacking enzymatic activity. Three of the four mutants have molecular weights similar to wild type enzyme and maintained an ability to bind GDP, whereas the other (Cys(104)) produced a series of lower molecular weight bands when characterized by Western blot analysis, and did not bind GDP. FucTs have highly conserved, potential N-linked sites, and our mass spectrometry analyses demonstrated that both N-linked sites are modified with oligosaccharides. PMID- 10816555 TI - Deletion of the serotonin 5-HT2C receptor PDZ recognition motif prevents receptor phosphorylation and delays resensitization of receptor responses. AB - Phosphorylation-deficient serotonin 5-HT(2C) receptors were generated to determine whether phosphorylation promotes desensitization of receptor responses. Phosphorylation of mutant 5-HT(2C) receptors that lack the carboxyl-terminal PDZ recognition motif (Ser(458)-Ser-Val-COOH; DeltaPDZ) was not detectable based on a band-shift phosphorylation assay and incorporation of (32)P. Treatment of cells stably expressing DeltaPDZ or wild-type 5-HT(2C) receptors with serotonin produced identical maximal responses and EC(50) values for eliciting [(3)H]inositol phosphate formation. In calcium imaging studies, treatment of cells expressing DeltaPDZ or wild-type 5-HT(2C) receptors with 100 nm serotonin elicited initial maximal responses and decay rates that were indistinguishable. However, a second application of serotonin 2.5 min after washout caused maximal responses that were approximately 5-fold lower with DeltaPDZ receptors relative to wild-type 5-HT(2C) receptors. After 10 min, responses of DeltaPDZ receptors recovered to wild-type 5-HT(2C) receptor levels. Receptors with single mutations at Ser(458) (S458A) or Ser(459) (S459A) decreased serotonin-mediated phosphorylation to 50% of wild-type receptor levels. Furthermore, subsequent calcium responses of S459A receptors were diminished relative to S458A and wild type receptors. These results establish that desensitization occurs in the absence of 5-HT(2C) receptor phosphorylation and suggest that receptor phosphorylation at Ser(459) enhances resensitization of 5-HT(2C) receptor responses. PMID- 10816556 TI - Close approximation of putative alpha -helices II, IV, VII, X, and XI in the translocation pathway of the lactose transport protein of Streptococcus thermophilus. AB - The lactose transport protein (LacS) of Streptococcus thermophilus belongs to a family of transporters in which putative alpha-helices II and IV have been implicated in cation binding and the coupled transport of the substrate and the cation. Here, the analysis of site-directed mutants shows that a positive and negative charge at positions 64 and 71 in helix II are essential for transport, but not for lactose binding. The conservation of charge/side-chain properties is less critical for Glu-67 and Ile-70 in helix II, and Asp-133 and Lys-139 in helix IV, but these residues are important for the coupled transport of lactose together with a proton. The analysis of second-site suppressor mutants indicates an ion pair exists between helices II and IV, and thus a close approximation of these helices can be made. The second-site suppressor analysis also suggests ion pairing between helix II and the intracellular loops 6-7 and 10-11. Because the C terminal region of the transmembrane domain, especially helix XI and loop 10-11, is important for substrate binding in this family of proteins, we propose that sugar and proton binding and translocation are performed by the joint action of these regions in the protein. Indeed, substrate protection of maleimide labeling of single cysteine mutants confirms that alpha-helices II and IV are directly interacting or at least conformationally involved in sugar binding and/or translocation. On the basis of new and published data, we reason that the helices II, IV, VII, X, and XI and the intracellular loops 6-7 and 10-11 are in close proximity and form the binding sites and/or the translocation pathway in the transporters of the galactosides-pentosides-hexuronides family. PMID- 10816557 TI - An essential role of the nuclear factor of activated T cells in the regulation of the expression of the cyclooxygenase-2 gene in human T lymphocytes. AB - We have previously reported that transcriptional induction of cyclooxygenase-2 (COX-2) isoenzyme occurs early after T cell receptor triggering, suggesting functional implications of cyclooxygenase activity in this process. Here, we identify the cis-acting elements responsible for the transcriptional activation of this gene in human T lymphocytes. COX-2 promoter activity was induced upon T cell activation both in primary resting T lymphocytes and in Jurkat cells. This induction was abrogated by inhibition of calcineurin phosphatase with the immunosuppressive drug cyclosporin A, whereas expression of an active calcineurin catalytic subunit enhanced COX-2 transcriptional activation. Moreover, cotransfection of nuclear factor of activated T cells (NFAT) wild type protein transactivated COX-2 promoter activity. Conversely, dominant negative mutants of NFATc or c-Jun proteins inhibited COX-2 induction. Electrophoretic mobility shift assays and site-directed mutagenesis allowed the identification of two regions of DNA located in the positions -117 and -58 relative to the transcriptional start site that serves as NFAT recognition sequences. These results emphasize the central role that the Ca(2+)/calcineurin pathway plays in COX-2 transcriptional regulation in T lymphocytes pointing to NFAT/activator protein-1 transcription factors as essential for COX-2 promoter regulation in these cells. PMID- 10816558 TI - The survival motor neuron protein of Schizosacharomyces pombe. Conservation of survival motor neuron interaction domains in divergent organisms. AB - Spinal muscular atrophy is a common often lethal neurodegenerative disease resulting from deletions or mutations in the survival motor neuron gene (SMN). SMN is ubiquitously expressed in metazoan cells and plays a role in small nuclear ribonucleoprotein assembly and pre-mRNA splicing. Here we characterize the Schizosacharomyces pombe orthologue of SMN (yeast SMN (ySMN)). We report that the ySMN protein is essential for viability and localizes in both the cytoplasm and the nucleus. Like human SMN, we show that ySMN can oligomerize. Remarkably, ySMN interacts directly with human SMN and Sm proteins. The highly conserved carboxyl terminal domain of ySMN is necessary for the evolutionarily conserved interactions of SMN and required for cell viability. We also demonstrate that the conserved amino-terminal region of ySMN is not required for SMN and Sm binding but is critical for the housekeeping function of SMN. PMID- 10816559 TI - The docking of primed vacuoles can be reversibly arrested by excess Sec17p (alpha SNAP). AB - Homotypic vacuole fusion occurs in ordered stages of priming, docking, and fusion. Priming, which prepares vacuoles for productive association, requires Sec17p (the yeast homolog of alpha-SNAP), Sec18p (the yeast NSF, an ATP-driven chaperone), and ATP. Sec17p is initially an integral part of the cis-SNARE complex together with vacuolar SNARE proteins and Sec18p (NSF). Previous studies have shown that Sec17p is rapidly released from the vacuole membrane during priming as the cis-SNARE complex is disassembled, but the order and causal relationship of these subreactions has not been known. We now report that the addition of excess recombinant his(6)-Sec17p to primed vacuoles can block subsequent docking. This inhibition is reversible by Sec18p, but the reaction cannot proceed to the tethering and trans-SNARE pairing steps of docking while the Sec17p block is in place. Once docking has occurred, excess Sec17p does not inhibit membrane fusion per se. Incubation of cells with thermosensitive Sec17-1p at nonpermissive temperature causes SNARE complex disassembly. These data suggest that Sec17p can stabilize vacuolar cis-SNARE complexes and that the release of Sec17p by Sec18p and ATP allows disassembly of this complex and activates its components for docking. PMID- 10816560 TI - Ligand interactions in the adenosine nucleotide-binding domain of the Hsp90 chaperone, GRP94. II. Ligand-mediated activation of GRP94 molecular chaperone and peptide binding activity. AB - The N-terminal domain of eukaryotic Hsp90 proteins contains a conserved adenosine nucleotide binding pocket that also serves as the binding site for the Hsp90 inhibitors geldanamycin and radicicol. Although this domain is essential for Hsp90 function, the molecular basis for adenosine nucleotide-dependent regulation of GRP94, the endoplasmic reticulum paralog of Hsp90, remains to be established. We report that bis-ANS (1,1'-bis(4-anilino-5-napthalenesulfonic acid), an environment sensitive fluorophore known to interact with nucleotide-binding domains, binds to the adenosine nucleotide-binding domain of GRP94 and thereby activates its molecular chaperone and peptide binding activities. bis-ANS was observed to elicit a tertiary conformational change in GRP94 similar to that occurring upon heat shock, which also activates GRP94 function. bis-ANS activation of GRP94 function was efficiently blocked by radicicol, an established inhibitory ligand for the adenosine nucleotide binding pocket. Confirmation of the N-terminal nucleotide binding pocket as the bis-ANS-binding site was obtained following covalent incorporation of bis-ANS into GRP94, trypsinolysis, and sequencing of bis-ANS-labeled limit digestion products. These data identify a ligand dependent regulation of GRP94 function and suggest a model whereby GRP94 function is regulated through a ligand-dependent conversion of GRP94 from an inactive to an active conformation. PMID- 10816561 TI - Ligand interactions in the adenosine nucleotide-binding domain of the Hsp90 chaperone, GRP94. I. Evidence for allosteric regulation of ligand binding. AB - X-ray crystallographic studies of the N-terminal domain of Hsp90 have identified an unconventional ATP binding fold, thereby inferring a role for ATP in the regulation of the Hsp90 activity. In this report, N-ethylcarboxamidoadenosine (NECA) was used to investigate the nucleotide binding properties of GRP94, the endoplasmic reticulum paralog of Hsp90. Whereas Hsp90 did not bind NECA, GRP94 bound NECA in a saturable manner with a K(d) of 200 nm. NECA binding to GRP94 was efficiently blocked by geldanamycin and radicicol. Analysis of ligand binding stoichiometries by radioligand and calorimetric techniques indicated that GRP94 bound 1 mol of NECA/mol of GRP94 dimer. In contrast, GRP94 bound radicicol at a stoichiometry of 2 mol of radicicol/mol of GRP94 dimer. In [(3)H]NECA displacement assays, GRP94 displayed binding interactions with ATP, dATP, ADP, AMP, cAMP, and adenosine, but not GTP, CTP, or UTP. To accommodate the 0.5 mol of NECA:mol of GRP94 binding stoichiometry observed for the native GRP94 dimer, a model for allosteric regulation (negative cooperativity) of ligand binding is proposed. A hypothesis on the regulation of GRP94 conformation and activity by adenosine-based ligand(s) other than ATP and ADP is presented. PMID- 10816562 TI - The C-terminal region of proSAAS is a potent inhibitor of prohormone convertase 1. AB - ProSAAS is a recently discovered 26-kDa neuroendocrine protein that was previously found to inhibit prohormone convertase (PC) 1 and not PC2. In the present study, the specificity of proSAAS toward other members of the prohormone convertase family was determined. Two microm proSAAS selectively inhibits PC1 but not furin, PACE4, PC5A, or PC7. The PC1 inhibitory region of proSAAS was mapped to an 8-12-residue region near the C terminus that includes a critical Lys-Arg sequence. Synthetic peptides corresponding to this region are competitive inhibitors of PC1 with apparent K(i) values of 14-40 nm. The inhibition becomes more effective with incubation time, indicating that the inhibitor is slow binding. A fusion protein containing the inhibitory region of proSAAS linked to the C terminus of glutathione S-transferase binds the 71-kDa form but not the 85 kDa form of PC1. This binding, which occurs at pH 5.5 and not at pH 7.4, is stable to incubation at room temperature for 1 h in the presence or absence of 0.5% Triton X-100 and/or 0.5 m NaCl. The removal of Ca(2+) with chelating agents partially releases the bound PC1. High concentrations of the inhibitory peptide quantitatively release the bound PC1. Taken together, these data support the proposal that proSAAS functions as an endogenous inhibitor of PC1. PMID- 10816563 TI - Immediate-early MEK-1-dependent stabilization of rat smooth muscle cell cyclooxygenase-2 mRNA by Galpha(q)-coupled receptor signaling. AB - Activation of Galpha(q)-coupled P2Y nucleotide receptors strongly (>100-fold) induces the rat vascular smooth muscle cell cyclooxygenase-2 (COX-2) mRNA, yet transcription is induced only approximately 3-fold over 1 h. Intact cell decay analysis of tetracycline-suppressible luciferase chimera mRNAs shows that regulated stabilization of the intrinsically unstable mRNA contributes to this response. Deletion mapping of the 2468-base COX-2 mRNA 3'-untranslated region (UTR) shows that a distal, 130-base AU-rich region functions as a cis-acting regulated stabilization response element, which under basal conditions serves as the dominant instability determinant for the 3'-UTR. Regulation of this response is through the p42/44 MAP kinases, whereas the p38 MAP kinases are not involved. The stabilization response element binds avidly and specifically to a prominent nuclear-enriched approximately 90-kDa factor and several less abundantly labeled mRNA binding proteins that are unaffected by P2Y receptor signaling. Although other instability determinants are located throughout the rat COX-2 mRNA 3'-UTR, mitogen signaling only interferes with rapid decay mediated by its most distal 130 bases. A complex of nuclear factors that bind this mRNA region specifically may include candidate targets for regulatory modulation. These observations support the general notion that the rapid induction of immediate-early gene expression through mitogenic receptors involves simultaneous activation of transcriptional and post-transcriptional mechanisms. PMID- 10816564 TI - Novel propeptide function in 20 S proteasome assembly influences beta subunit composition. AB - The assembly of eukaryotic 20 S proteasomes involves the formation of half proteasomes where precursor beta-type subunits gather in position on an alpha subunit ring, followed by the association of two half-proteasomes and beta subunit processing. In vertebrates three additional beta-subunits (beta1i/LMP2, beta2i/MECL1, and beta5i/LMP7) can be synthesized and substituted for constitutive homologues (beta1/delta, beta2/Z, and beta5/X) to yield immunoproteasomes, which are important for generating certain antigenic peptides. We have shown previously that when all six beta-subunits are present, cooperative assembly mechanisms limit the diversity of proteasome populations. Specifically, LMP7 is incorporated preferentially over X into preproteasomes containing LMP2 and MECL1. We show here that the LMP7 propeptide is responsible for this preferential incorporation, and it also enables LMP7 to incorporate into proteasomes containing delta and Z. In contrast, the X propeptide restricts incorporation to proteasomes with delta and Z. Furthermore, we demonstrate that the LMP7 propeptide can function in trans when expressed on LMP2, and that its NH(2)-terminal and mid-regions are particularly critical for function. In addition to identifying a novel propeptide function, our results raise the possibility that one consequence of LMP7 incorporation into both immunoproteasomes and delta/Z proteasomes may be to increase the diversity of antigenic peptides that can be generated. PMID- 10816565 TI - A Ca2+-dependent tryptic cleavage site and a protein kinase A phosphorylation site are present in the Ca2+ regulatory domain of scallop muscle Na+-Ca2+ exchanger. AB - Digestion of scallop muscle membrane fractions with trypsin led to release of soluble polypeptides derived from the large cytoplasmic domain of a Na(+)-Ca(2+) exchanger. In the presence of 1 mm Ca(2+), the major product was a peptide of approximately 37 kDa, with an N terminus corresponding to residue 401 of the NCX1 exchanger. In the presence of 10 mm EGTA, approximately 16- and approximately 19 kDa peptides were the major products. Polyclonal rabbit IgG raised against the 37 kDa peptide also bound to the 16- and 19-kDa soluble tryptic peptides and to a 105-110-kDa polypeptide in the undigested membrane preparation. The 16-kDa fragment corresponded to the N-terminal part of the 37-kDa peptide. The conformation of the precursor polypeptide chain in the region of the C terminus of the 16-kDa tryptic peptide was thus altered by the binding of Ca(2+). Phosphorylation of the parent membranes with the catalytic subunit of protein kinase A and [gamma-(32)P]ATP led to incorporation of (32)P into the 16- and 37 kDa soluble fragments. A site may exist within the Ca(2+) regulatory domain of a scallop muscle Na(+)-Ca(2+) exchanger that mediates direct modulation of secondary Ca(2+) regulation by cAMP. PMID- 10816566 TI - The zinc-responsive regulator Zur and its control of the znu gene cluster encoding the ZnuABC zinc uptake system in Escherichia coli. AB - The synthesis of the Escherichia coli zinc transporter, encoded by the znuACB gene cluster, is regulated in response to the intracellular zinc concentration by the zur gene product. Inactivation of the zur gene demonstrated that Zur acts as a repressor when binding Zn(2+). Eight chromosomal mutant zur alleles were sequenced to correlate the loss of Zur function with individual mutations. Wild type Zur and ZurDelta46-91 formed homo- and heterodimers. Dimerization was independent of metal ions since it also occurred in the presence of metal chelators. Using an in vivo titration assay, the znu operator was narrowed down to a 31-base pair region overlapping the translational start site of znuA. This location was confirmed by footprinting assays. Zur directly binds to a single region comprising a nearly perfect palindrome. Zinc chelators completely inhibited and Zn(2+) in low concentrations enhanced DNA binding of Zur. No evidence for autoregulation of Zur was found. Zur binds at least 2 zinc ions/dimer specifically. Although most of the mutant Zur proteins bound to the znu operator in vitro, no protection was observed in in vivo footprinting experiments. Analysis of the mutant Zur proteins suggested an amino-terminal DNA contact domain around residue 65 and a dimerization and Zn(2+)-binding domain toward the carboxyl-terminal end. PMID- 10816567 TI - Stimulation of human neutrophils by chemotactic factors is associated with the activation of phosphatidylinositol 3-kinase gamma. AB - The activation of human polymorphonuclear neutrophil leukocytes (neutrophils) is associated with an increased synthesis of the highly phosphorylated phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)). The aims of the present investigation were to determine whether the newly described, G protein dependent phosphatidylinositol 3-kinase (PI3K), p110gamma, was involved in the responses to chemotactic factors interacting with G protein-coupled receptors. The presence of p110gamma in neutrophils was first established both at the protein and the mRNA level. Stimulation of the cells with fMet-Leu-Phe or interleukin-8 increased the PI3K activity in p110gamma, but not p85, immunoprecipitates. The time course of this effect (threshold within less than 5 s, maximal activation at 10-15 s) was consistent with that of the generation of PtdIns(3,4,5)P(3). Wortmannin, a PI3K inhibitor, abrogated the effects of fMet Leu-Phe, which were also significantly inhibited by pertussis toxin. Finally, fMet-Leu-Phe also induced a significant translocation of p110gamma to a particulate fraction derived from these cells. These data indicate that p110gamma represent the major PI3K activated by fMet-Leu-Phe and interleukin-8 at very early time points following the stimulation of human neutrophils. PMID- 10816568 TI - Overexpression of SERCA2b in the heart leads to an increase in sarcoplasmic reticulum calcium transport function and increased cardiac contractility. AB - The sarcoplasmic reticulum calcium ATPase SERCA2b is an alternate isoform encoded by the SERCA2 gene. SERCA2b is expressed ubiquitously and has a higher Ca(2+) affinity compared with SERCA2a. We made transgenic mice that overexpress the rat SERCA2b cDNA in the heart. SERCA2b mRNA level was approximately approximately 20 fold higher than endogenous SERCA2b mRNA in transgenic hearts. SERCA2b protein was increased 8-10-fold in the heart, whereas SERCA2a mRNA/protein level remained unchanged. Confocal microscopy showed that SERCA2b is localized preferentially around the T-tubules of the SR, whereas SERCA2a isoform is distributed both transversely and longitudinally in the SR membrane. Calcium-dependent calcium uptake measurements showed that the maximal velocity of Ca(2+) uptake was not changed, but the apparent pump affinity for Ca(2+) (K(0.5)) was increased in SERCA2b transgenic mice (0.199 +/- 0.011 micrometer) compared with wild-type control mice (0.269 +/- 0.012 micrometer, p < 0.01). Work-performing heart preparations showed that SERCA2b transgenic hearts had a higher rates of contraction and relaxation, shorter time to peak pressure and half-time for relaxation than wild-type hearts. These data show that SERCA2b is associated in a subcompartment within the sarcoplasmic reticulum of cardiac myocytes. Overexpression of SERCA2b leads to an increase in SR calcium transport function and increased cardiac contractility, suggesting that SERCA2b plays a highly specialized role in regulating the beat-to-beat contraction of the heart. PMID- 10816569 TI - Conformational activation of radixin by G13 protein alpha subunit. AB - G(13) protein, one of the heterotrimeric guanine nucleotide-binding proteins (G proteins), regulates diverse and complex cellular responses by transducing signals from the cell surface presumably involving more than one pathway. Yeast two-hybrid screening of a mouse brain cDNA library identified radixin, a member of the ERM family of three closely related proteins (ezrin, radixin, and moesin), as a protein that interacted with Galpha(13). Interaction between radixin and Galpha(13) was confirmed by in vitro binding assay and by co-immunoprecipitation technique. Activated Galpha(13) induced conformational activation of radixin, as determined by binding of radixin to polymerized F-actin and by immunofluorescence in intact cells. Finally, two dominant negative mutants of radixin inhibited Galpha(13)-induced focus formation of Rat-1 fibroblasts but did not affect Ras induced focus formation. Our results identifying a new signaling pathway for Galpha(13) indicate that ERM proteins can be activated by and serve as effectors of heterotrimeric G proteins. PMID- 10816570 TI - A possible role for caveolin as a signaling organizer in olfactory sensory membranes. AB - Fast kinetics and sensitivity of olfactory signaling raise the question of whether the participating proteins may be associated in supramolecular transduction complexes. We found evidence that caveolin proteins could play an important role in organizing signaling elements in olfactory sensory neurons. Western blot analysis indicated that caveolins are highly enriched in olfactory sensory membranes, where they co-localize in detergent-insoluble complexes with key components of the signaling pathways. Furthermore, the results of immunoprecipitation experiments suggest that G proteins and effector enzyme form preassembled subcellular complexes with caveolins. Since anti-caveolin antibodies and synthetic peptides derived from the scaffolding domains of caveolin-1 and caveolin-2 effectively attenuated second messenger responses in sensory cilia preparations in a characteristic manner, the data led to the suggestion that caveolins could mediate the assembly of signaling complexes within specialized membrane microdomains of olfactory sensory neurons. PMID- 10816571 TI - Interaction between endothelial differentiation-related factor-1 and calmodulin in vitro and in vivo. AB - Calmodulin (CaM) is the principal Ca(2+) receptor protein inside the cell. When activated by Ca(2+), CaM binds and activates target proteins, thus altering the metabolism and physiology of the cell. Under basal conditions, calcium-free CaM binds to other proteins termed CaM-binding proteins. Recently, we described endothelial differentiation-related factor (EDF)-1 as a protein involved in the repression of endothelial cell differentiation (Dragoni, I., Mariotti, M., Consalez, G. G., Soria, M., and Maier, J. A. M. (1998) J. Biol. Chem. 273, 31119 31124). Here we report that (i) EDF-1 binds CaM in vitro and in vivo; (ii) EDF-1 is phosphorylated in vitro and in vivo by protein kinase C; and (iii) EDF-1-CaM interaction is modulated by the concentrations of Ca(2+) and by the phosphorylation of EDF-1 by protein kinase C both in vitro and in vivo. In addition, 12-O-tetradecanoylphorbol-13-acetate treatment of human umbilical vein endothelial cell stimulates the nuclear translocation of EDF-1. On the basis of the high homology of EDF-1 with multiprotein bridging factor-1, a transcriptional coactivator that binds TATA-binding protein (TBP), we also demonstrate that EDF-1 interacts with TBP in vitro and in human endothelial cells. We hypothesize that EDF-1 serves two main functions in endothelial cells as follows: (i) to bind CaM in the cytosol at physiologic concentrations of Ca(2+) and (ii) to act in the nucleus as a transcriptional coactivator through its binding to TBP. PMID- 10816572 TI - Caveolin-1 expression inhibits Wnt/beta-catenin/Lef-1 signaling by recruiting beta-catenin to caveolae membrane domains. AB - Caveolin-1 is a principal component of caveolae membranes. In NIH 3T3 cells, caveolin-1 expression is dramatically up-regulated in confluent cells and localizes at areas of cell-cell contact. However, it remains unknown whether caveolin-1 is involved in cell-cell signaling. Here, we examine the potential role of caveolin-1 in regulating beta-catenin signaling. beta-Catenin plays a dual role in the cell, linking E-cadherin to the actin cytoskeleton and in Wnt signaling by forming a complex with members of the lymphoid enhancing factor (Lef 1) family of transcription factors. We show that E-cadherin, beta-catenin, and gamma-catenin (plakoglobin) are all concentrated in caveolae membranes. Moreover, we demonstrate that activation of beta-catenin/Lef-1 signaling by Wnt-1 or by overexpression of beta-catenin itself is inhibited by caveolin-1 expression. We also show that recombinant expression of caveolin-1 in caveolin-1 negative cells is sufficient to recruit beta-catenin to caveolae membranes, thereby blocking beta-catenin-mediated transactivation. These results suggest that caveolin-1 expression can modulate Wnt/beta-catenin/Lef-1 signaling by regulating the intracellular localization of beta-catenin. PMID- 10816573 TI - Human DnaJ homologs dj2 and dj3, and bag-1 are positive cochaperones of hsc70. AB - DnaJ is an essential cochaperone of mammalian heat shock cognate 70 (hsc70) protein. We previously found that dj2 (HSDJ/hdj-2/rdj1), rather than dj1 (hsp40/hdj-1), is a partner DnaJ for the hsc70-based chaperone system. Here, we compared the distribution of dj1, dj2, and the newly found dj3 (cpr3/DNJ3/HIRIP4/rdj2) in cultured cells. Both dj3 as well as dj2 were farnesylated and were ubiquitously expressed. In immunocytochemical and subfractionation studies, these two proteins colocalized with hsc70 under normal conditions. However, dj1 and hsc70 apparently colocalized in the nucleoli after heat shock. Simultaneous depletion of dj2 and dj3 from rabbit reticulocyte lysate markedly reduced mitochondrial import of pre-ornithine transcarbamylase and refolding of guanidine-denatured luciferase. Re-addition of either dj2 or dj3 led to recovery of these reactions. In a reconstituted system, both hsc70-dj2 and hsc70-dj3 were effective in protein refolding. Anti-apoptotic protein bag-1 further stimulated ATP hydrolysis and protein refolding by both pairs. Thus, dj2 and dj3 are the partner DnaJs of hsc70 within the cell, functionally similar and much more efficient than dj1, and bag-1 is a positive cochaperone of the hsc70 dj2 and hsc70-dj3 systems. PMID- 10816574 TI - Absence of the mitochondrial AAA protease Yme1p restores F0-ATPase subunit accumulation in an oxa1 deletion mutant of Saccharomyces cerevisiae. AB - The nuclear gene OXA1 encodes a protein located within the mitochondrial inner membrane that is required for the biogenesis of both cytochrome c oxidase (Cox) and ATPase. In the absence of Oxa1p, the translocation of the mitochondrially encoded subunit Cox2p to the intermembrane space (also referred to as export) is prevented, and it has been proposed that Oxa1p could be a component of a general mitochondrial export machinery. We have examined the role of Oxa1p in light of its relationships with two mitochondrial proteases, the matrix protease Afg3p Rca1p and the intermembrane space protease Yme1p, by analyzing the assembly and activity of the Cox and ATPase complexes in Deltaoxa1, Deltaoxa1Deltaafg3, and Deltaoxa1Deltayme1 mutants. We show that membrane subunits of both complexes are specifically degraded in the absence of Oxa1p. Neither Afg3p nor Yme1p is responsible for the degradation of Cox subunits. However, the F(0) subunits Atp4p, Atp6p, and Atp17p are stabilized in the Deltaoxa1Deltayme1 double mutant, and oligomycin-sensitive ATPase activity is restored, showing that the increased stability of the ATPase subunits allows significant translocation and assembly to occur even in the absence of Oxa1p. These results suggest that Oxa1p is not essential for the export of ATPase subunits. In addition, although respiratory function is dispensable in Saccharomyces cerevisiae, we show that the simultaneous inactivation of AFG3 and YME1 is lethal and that the essential function does not reside in their protease activity. PMID- 10816575 TI - Inhibition of vascular endothelial growth factor expression in HEC1A endometrial cancer cells through interactions of estrogen receptor alpha and Sp3 proteins. AB - Treatment of HEC1A endometrial cancer cells with 10 nm 17beta-estradiol (E2) resulted in decreased vascular endothelial growth factor (VEGF) mRNA expression, and a similar response was observed using a construct, pVEGF1, containing a VEGF gene promoter insert from -2018 to +50. In HEC1A cells transiently transfected with pVEGF1 and a series of deletion plasmids, it was shown that E2-dependent down-regulation was dependent on wild-type estrogen receptor alpha (ERalpha) and reversed by the anti-estrogen ICI 182, 780, and this response was not affected by progestins. Deletion analysis of the VEGF gene promoter identified an overlapping G/GC-rich site between -66 to -47 that was required for decreased transactivation by E2. Protein-DNA binding studies using electrophoretic mobility shift and DNA footprinting assays showed that both Sp1 and Sp3 proteins bound this region of the VEGF promoter. Coimmunoprecipitation and pull-down assays demonstrated that Sp3 and ERalpha proteins physically interact, and the interacting domains of both proteins are different from those previously observed for interactions between Sp1 and ERalpha proteins. Using a dominant negative form of Sp3 and transcriptional activation assays in Schneider SL-2 insect cells, it was confirmed that ERalpha-Sp3 interactions define a pathway for E2-mediated inhibition of gene expression, and this represents a new mechanism for decreased gene expression by E2. PMID- 10816576 TI - Threonine phosphorylation diverts internalized epidermal growth factor receptors from a degradative pathway to the recycling endosome. AB - Transregulation of the epidermal growth factor receptor (EGFR) by protein kinase C (PKC) serves as a model for heterologous desensitization of receptor tyrosine kinases, but the underlying mechanism remained unknown. By using c-Cbl-induced ubiquitination of EGFR as a marker for transfer from early to late endosomes, we provide evidence that PKC can inhibit this process. In parallel, receptor down regulation and degradation are significantly reduced. The inhibitory effects of PKC are mediated by a single threonine residue (threonine 654) of EGFR, which serves as a major PKC phosphorylation site. Biochemical and morphological analyses indicate that threonine-phosphorylated EGFR molecules undergo normal internalization, but instead of sorting to lysosomal degradation, they recycle back to the cell surface. In conclusion, by sorting EGFR to the recycling endosome, heterologous desensitization restrains ligand-induced down-regulation of EGFR. PMID- 10816577 TI - Characterization of isolated acidocalcisomes of Trypanosoma cruzi. AB - The acidocalcisome is an acidic calcium store in trypanosomatids with a vacuolar type proton-pumping pyrophosphatase (V-H(+)-PPase) located in its membrane. In this paper, we describe a new method using iodixanol density gradients for purification of the acidocalcisome from Trypanosoma cruzi epimastigotes. Pyrophosphatase assays indicated that the isolated organelle was at least 60-fold purified compared with the large organelle (10,000 x g) fraction. Assays for other organelles generally indicated no enrichment in the acidocalcisome fraction; glycosomes were concentrated 5-fold. Vanadate-sensitive ATP-driven Ca(2+) uptake (Ca(2+)-ATPase) activity was detectable in the isolated acidocalcisome, but ionophore experiments indicated that it was not acidic. However, when pyrophosphate was added, the organelle acidified, and the rate of Ca(2+) uptake increased. Use of the indicator Oxonol VI showed that V-H(+)-PPase activity generated a membrane potential. Use of sulfate or nitrate in place of chloride in the assay buffer did not affect V-H(+)-PPase activity, but there was less activity with gluconate. Organelle acidification was countered by the chloride/proton symport cycloprogidiosin. No vacuolar H(+)-ATPase activity was detectable in isolated acidocalcisomes. However, immunoblots showed the presence of at least a membrane-bound V-H(+)-ATPase subunit, while experiments employing permeabilized epimastigotes suggested that vacuolar H(+)-ATPase and V-H(+)-PPase activities are present in the same Ca(2+)-containing compartment. PMID- 10816578 TI - Requirement of an AP-1 site in the calcium response region of the involucrin promoter. AB - Involucrin is a major protein of the cornified envelope of keratinocytes that provides much of the structural integrity of the skin. The gene expression of this differentiation marker is induced by elevated extracellular calcium in cultured human keratinocytes. A 3.7-kilobase fragment of this gene contains the necessary elements to drive a luciferase reporter in a calcium-dependent manner. We have sequenced the upstream region of the involucrin promoter and localized a calcium response element that contains an activating protein-1 (AP-1) site (TGAGTCA). Mutation of this site abolished the promoter activation by calcium. Compared with cells grown in 0.03 mm calcium, the binding activity of factors within nuclear extracts from keratinocytes for this AP-1 site was enhanced 3-fold in cells grown in 1.2 mm calcium. Immunoelectrophoretic mobility shift (supershift) assays identified JunD, Fra1, and Fra2 as the major factors that bind to the AP-1 element. Western analysis of the proteins in the nuclear extracts showed that the levels of c-Jun, JunB, JunD, FosB, and Fra2 increased and the levels of c-Fos and Fra1 decreased slightly with calcium treatment. The effect of calcium on the involucrin promoter was enhanced synergistically by phorbol 12-myristate 13-acetate (PMA) in a protein kinase-dependent manner. In conclusion, calcium-regulated involucrin gene expression is mediated at least in part by AP-1 transcription factors. PMID- 10816579 TI - Control of bisecting GlcNAc addition to N-linked sugar chains. AB - In the present study, experimental control of the formation of bisecting GlcNAc was investigated, and the competition between beta-1,4-GalT (UDP-galactose:N acetylglucosamine beta-1, 4-galactosyltransferase) and GnT-III (UDP-N acetylglucosamine:beta-d-mannoside beta-1, 4-N-acetylglucosaminyltransferase) was examined. We isolated a beta-1,4-GalT-I single knockout human B cell clone producing monoclonal IgM and several transfectant clones that overexpressed beta 1,4-GalT-I or GnT-III. In the beta-1,4-GalT-I-single knockout cells, the extent of bisecting GlcNAc addition to the sugar chains of IgM was increased, where beta 1,4-GalT activity was reduced to about half that in the parental cells, and GnT III activity was unaltered. In the beta-1,4-GalT-I transfectants, the extent of bisecting GlcNAc addition was reduced although GnT-III activity was not altered significantly. In the GnT-III transfectants, the extent of bisecting GlcNAc addition increased along with the increase in levels of GnT-III activity. The extent of bisecting GlcNAc addition to the sugar chains of IgM was significantly correlated with the level of intracellular beta-1,4-GalT activity relative to that of GnT-III. These results were interpreted as indicating that beta-1, 4-GalT competes with GnT-III for substrate in the cells. PMID- 10816580 TI - Characterization of glucosinolate uptake by leaf protoplasts of Brassica napus. AB - The uptake of radiolabeled p-hydroxybenzylglucosinolate (p-OHBG) by protoplasts isolated from leaves of Brassica napus was detected using silicone oil filtration technique. The uptake was pH-dependent with higher uptake rates at acidic pH. Imposition of a pH gradient (internal alkaline) across the plasma membrane resulted in a rapid uptake of p-OHBG, which was inhibited in the presence of carbonyl cyanide m-chlorophenylhydrazone, indicating that the uptake is dependent on a proton motive force. Dissipation of the internal positive membrane potential generated a small influx as compared with that seen for pH gradient (DeltapH). Kinetic studies demonstrated the presence of two uptake systems, a saturable and a linear component. The saturable kinetics indicated carrier-mediated translocation with a K(m) of 1.0 mm and a V(max) of 28.7 nmol/microl/h. The linear component had very low substrate affinity. The carrier-mediated transport had a temperature coefficient (Q(10)) of 1.8 +/- 0.2 in the temperature range from 4-30 degrees C. The uptake was against a concentration gradient and was sensitive to protonophores, uncouplers, H(+)-ATPase inhibitors, and the sulfhydryl group modifier p-chloromercuriphenylsulfonic acid. The carrier mediated uptake system had high specificity for glucosinolates because glucosinolate degradation products, amino acids, sugars, or glutathione conjugates did not compete for p-OHBG uptake. Glucosinolates with different side chains were equally good competitors of p-OHBG uptake, which indicates that the uptake system has low specificity for the glucosinolate side chains. Our data provide the first evidence of an active transport of glucosinolates by a proton coupled symporter in the plasma membrane of rape leaves. PMID- 10816581 TI - Determinants of translocation and folding of TreF, a trehalase of Escherichia coli. AB - One isoform of trehalase, TreF, is present in the cytoplasm and a second, TreA, in the periplasm. To study the questions of why one enzyme is exported efficiently and the other is not and whether these proteins can fold in their nonnative cellular compartment, we fused the signal sequence of periplasmic TreA to cytoplasmic TreF. Even though this TreF construct was exported efficiently to the periplasm, it was not active. It was insoluble and degraded by the periplasmic serine protease DegP. To determine why TreF was misfolded in the periplasm, we isolated and characterized Tre(+) revertants of periplasmic TreF. To further characterize periplasmic TreF, we used a genetic selection to isolate functional TreA-TreF hybrids, which were analyzed with respect to solubility and function. These data suggested that a domain located between residues 255 and 350 of TreF is sufficient to cause folding problems in the periplasm. In contrast to TreF, periplasmic TreA could fold into the active conformation in its nonnative cellular compartment, the cytoplasm, after removal of its signal sequence. PMID- 10816582 TI - FXXLF and WXXLF sequences mediate the NH2-terminal interaction with the ligand binding domain of the androgen receptor. AB - The nuclear receptor superfamily members of eukaryotic transcriptional regulators contain a highly conserved activation function 2 (AF2) in the hormone binding carboxyl-terminal domain and, for some, an additional activation function 1 in the NH(2)-terminal region which is not conserved. Recent biochemical and crystallographic studies revealed the molecular basis of AF2 is hormone-dependent recruitment of LXXLL motif-containing coactivators, including the p160 family, to a hydrophobic cleft in the ligand binding domain. Our previous studies demonstrated that AF2 in the androgen receptor (AR) binds only weakly to LXXLL motif-containing coactivators and instead mediates an androgen-dependent interaction with the AR NH(2)-terminal domain required for its physiological function. Here we demonstrate in a mammalian two-hybrid assay, glutathione S transferase fusion protein binding studies, and functional assays that two predicted alpha-helical regions that are similar, but functionally distinct from the p160 coactivator interaction sequence, mediate the androgen-dependent, NH(2)- and carboxyl-terminal interaction. FXXLF in the AR NH(2)-terminal domain with the sequence (23)FQNLF(27) mediates interaction with AF2 and is the predominant androgen-dependent interaction site. This FXXLF sequence and a second NH(2) terminal WXXLF sequence (433)WHTLF(437) interact with different regions of the ligand binding domain to stabilize the hormone-receptor complex and may compete with AF2 recruitment of LXXLL motif-containing coactivators. The results suggest a unique mechanism for AR-mediated transcriptional activation. PMID- 10816583 TI - Presenilin 1 regulates pharmacologically distinct gamma -secretase activities. Implications for the role of presenilin in gamma -secretase cleavage. AB - Presenilins (PSs) are polytopic membrane proteins that have been implicated as potential therapeutic targets in Alzheimer's disease because of their role in regulating the gamma-secretase cleavage that generates the amyloid beta protein (Abeta). It is not clear how PSs regulate gamma-secretase cleavage, but there is evidence that PSs could be either essential cofactors in the gamma-secretase cleavage, gamma-secretase themselves, or regulators of intracellular trafficking that indirectly influence gamma-secretase cleavage. Using presenilin 1 (PS1) mutants that inhibit Abeta production in conjunction with transmembrane domain mutants of the amyloid protein precursor that are cleaved by pharmacologically distinct gamma-secretases, we show that PS1 regulates multiple pharmacologically distinct gamma-secretase activities as well as inducible alpha-secretase activity. It is likely that PS1 acts indirectly to regulate these activities (as in a trafficking or chaperone role), because these data indicate that for PS1 to be gamma-secretase it must either have multiple active sites or exist in a variety of catalytically active forms that are altered to an equivalent extent by the mutations we have studied. PMID- 10816584 TI - SPECs, small binding proteins for Cdc42. AB - The Rho GTPase, Cdc42, regulates a wide variety of cellular activities including actin polymerization, focal complex assembly, and kinase signaling. We have identified a new family of very small Cdc42-binding proteins, designated SPECs (for Small Protein Effector of Cdc42), that modulates these regulatory activities. The two human members, SPEC1 and SPEC2, encode proteins of 79 and 84 amino acids, respectively. Both contain a conserved N-terminal region and a centrally located CRIB (Cdc42/Rac Interactive Binding) domain. Using a yeast two hybrid system, we found that both SPECs interact strongly with Cdc42, weakly with Rac1, and not at all with RhoA. Transfection analysis revealed that SPEC1 inhibited Cdc42-induced c-Jun N-terminal kinase (JNK) activation in COS1 cells in a manner that required an intact CRIB domain. Immunofluorescence experiments in NIH-3T3 fibroblasts demonstrated that both SPEC1 and SPEC2 showed a cortical localization and induced the formation of cell surface membrane blebs, which was not dependent on Cdc42 activity. Cotransfection experiments demonstrated that SPEC1 altered Cdc42-induced cell shape changes both in COS1 cells and in NIH-3T3 fibroblasts and that this alteration required an intact CRIB domain. These results suggest that SPECs act as novel scaffold molecules to coordinate and/or mediate Cdc42 signaling activities. PMID- 10816585 TI - Cross-linking of plasminogen activator inhibitor 2 and alpha 2-antiplasmin to fibrin(ogen). AB - In this study, we identified lysine residues in the fibrinogen Aalpha chain that serve as substrates during transglutaminase (TG)-mediated cross-linking of plasminogen activator inhibitor 2 (PAI-2). Comparisons were made with alpha(2) antiplasmin (alpha(2)-AP), which is known to cross-link to lysine 303 of the Aalpha chain. A 30-residue peptide containing Lys-303 specifically competed with fibrinogen for cross-linking to alpha(2)-AP but not for cross-linking to PAI-2. Further evidence that PAI-2 did not cross-link via Lys-303 was the cross-linking of PAI-2 to I-9 and des-alphaC fibrinogens, which lack 100 and 390 amino acids from the C terminus of the Aalpha chain, respectively. PAI-2 or alpha(2)-AP was cross-linked to fibrinogen and digested with trypsin or endopeptidase Glu-C, and the resulting peptides analyzed by mass spectrometry. Peptides detected were consistent with tissue TG (tTG)-mediated cross-linking of PAI-2 to lysines 148, 176, 183, 457 and factor XIIIa-mediated cross-linking of PAI-2 to lysines 148, 230, and 413 in the Aalpha chain. alpha(2)-AP was cross-linked only to lysine 303. Cross-linking of PAI-2 to fibrinogen did not compete with alpha(2)-AP, and the two proteins utilized different lysines in the Aalpha chain. Therefore, PAI-2 and alpha(2)-AP can cross-link simultaneously to the alpha polymers of a fibrin clot and promote resistance to lysis. PMID- 10816586 TI - Membrane association of and critical residues in the catalytic domain of human neuropathy target esterase. AB - Neuropathy target esterase (NTE) is an integral membrane protein in vertebrate neurons and a member of a novel family of putative serine hydrolases. Here we show that NEST, a recombinant polypeptide expressed in Escherichia coli, reacts with an ester substrate and covalent inhibitors in a manner very similar to NTE. NEST comprises residues 727-1216 of human NTE, and site-directed mutagenesis revealed that serine 966 and two aspartate residues, Asp(1086) and Asp(960), are critical for catalysis. The results of mutating the 11 histidines in NEST suggest that NTE does not use a conventional catalytic triad. By reacting NEST with [(3)H]diisopropyl fluorophosphate, Ser(966) was confirmed as the active-site serine, and evidence was obtained that an isopropyl group is transferred from the Ser(966) adduct to an aspartate residue. Detergent was required both for solubilization of NEST from lysates of E. coli and during purification procedures. Catalytic activity was lost in detergent extracts, but was restored when purified NEST was incorporated into dioleoylphosphatidylcholine liposomes. Hydropathy analysis did not indicate the presence of membrane-spanning segments within the NEST sequence. However, biochemical evidence including detergent-phase separation experiments and the resistance of liposome-incorporated NEST to proteolysis indicated that, unlike most eukaryotic serine hydrolases, the catalytic domain of NTE has integral membrane protein properties. PMID- 10816587 TI - Identification of a short (C15) chain Z-isoprenyl diphosphate synthase and a homologous long (C50) chain isoprenyl diphosphate synthase in Mycobacterium tuberculosis. AB - We report the cloning, overexpression, and partial characterization of two unique Z-isoprenyl diphosphate synthase homologs from Mycobacterium tuberculosis. The first enzyme, Rv1086, adds one isoprene unit to omega,E-geranyl diphosphate. The product, omega,E, Z-farnesyl diphosphate, is the putative substrate of the second enzyme, Rv2361c. This enzyme adds seven more isoprene units to omega, E,Z farnesyl diphosphate and releases decaprenyl diphosphate. Both open reading frames were cloned from the M. tuberculosis H37Rv genome and overexpressed in M. smegmatis. Membrane and cytosol fractions from wild type and the two recombinant strains were assayed for [(14)C]isopentenyl diphosphate incorporation into isoprenyl diphosphates in the presence of various allylic isoprenyl diphosphate acceptors. Membrane fractions of recombinant cells overexpressing Rv2361c incubated with farnesyl diphosphate showed a 10-fold increase of [(14)C]isopentenyl diphosphate incorporation into decaprenyl diphosphate. Membrane fractions of recombinant cells overexpressing Rv1086 incubated with geranyl diphosphate showed a 5-fold increase of [(14)C]isopentenyl diphosphate incorporation into farnesyl diphosphate. Analysis of the stereochemistry revealed that all of the overexpressed farnesyl diphosphate was in the omega,E, Z configuration. This is the first description of a short chain isoprenyl diphosphate synthase that generates products with Z-stereochemistry. Previously, all known short chain isoprenyl diphosphate synthases catalyze the synthesis of products with E-stereochemistry. PMID- 10816588 TI - KCNQ5, a novel potassium channel broadly expressed in brain, mediates M-type currents. AB - KCNQ2 and KCNQ3, both of which are mutated in a type of human neonatal epilepsy, form heteromeric potassium channels that are expressed in broad regions of the brain. The associated current may be identical to the M-current, an important regulator of neuronal excitability. We now show that the RNA encoding the novel KCNQ5 channel is also expressed in brain and in sympathetic ganglia where it overlaps largely with KCNQ2 and KCNQ3. In addition, it is expressed in skeletal muscle. KCNQ5 yields currents that activate slowly with depolarization and can form heteromeric channels with KCNQ3. Currents expressed from KCNQ5 have voltage dependences and inhibitor sensitivities in common with M-currents. They are also inhibited by M1 muscarinic receptor activation. A KCNQ5 splice variant found in skeletal muscle displays altered gating kinetics. This indicates a molecular diversity of channels yielding M-type currents and suggests a role for KCNQ5 in the regulation of neuronal excitability. PMID- 10816589 TI - Cobalt-mediated dimerization of the human natural killer cell inhibitory receptor. AB - Upon engagement of specific class I major histocompatibility complex (MHC) molecules on target cells, inhibitory receptors on natural killer (NK) cells deliver a negative signal that prevents the target cell lysis by NK cells. In humans, killer cell immunoglobulin-related receptors (KIR) with two immunoglobulin-like domains (KIR2D) modulate the lysis of target cells bearing specific HLA-C alleles (Moretta, A., Vitale, M., Bottino, C., Orengo, A. M., Morelli, L., Augugliaro, R., Barbaresi, M., Ciccone, E., and Moretta, L. (1993) J. Exp. Med. 178, 597-604). The transduction of inhibitory signals by KIR2D molecules is impaired by the zinc chelator, 1,10-phenanthroline, and mutation of a putative zinc-binding site (Rajagopalan, S., and Long, E. O. (1998) J. Immunol. 161, 1299-1305), but the mechanism by which zinc may affect the function of KIR remains unknown. In this study, the inhibitory NK receptor KIR2DL1 was discovered to dimerize in the presence of Co(2+) as observed on native gel electrophoresis and by gel filtration column chromatography. Furthermore, Co(2+)-mediated KIR2DL1 dimer binds to HLA-Cw4 with higher affinity than the wild type KIR2DL1 monomer. Replacement of the amino-terminal His residue by Ala abolishes the ability of KIR2DL1 to bind Co(2+), indicating that Co(2+)-mediated KIR2DL1 dimerization involves pairing of the D1 domain. Although not observed on native gels, the inhibitory receptor KIR2DL1 can be chemically cross-linked into dimers in the presence of Zn(2+) and its related divalent metal ions, suggesting that Co(2+) mediated dimerization of KIR2DL1 may mimic a weaker interaction between KIR2DL1 and zinc in vivo. PMID- 10816590 TI - TRP4 (CCE1) protein is part of native calcium release-activated Ca2+-like channels in adrenal cells. AB - Mammalian TRP proteins have been implicated to function as ion channel subunits responsible for agonist-induced Ca(2+) entry. To date, TRP proteins have been extensively studied by heterologous expression giving rise to diverse channel properties and activation mechanisms including store-operated mechanisms. However, the molecular structure and the functional properties of native TRP channels still remain elusive. Here we analyze the properties of TRP4 (CCE1) channels in their native environment and characterize TRP expression patterns and store-operated calcium currents that are endogenous to bovine adrenal cells. We show by Northern blot analysis, immunoblots, and immunohistochemistry that TRP4 transcripts and TRP4 protein are present in the adrenal cortex but absent in the medulla. Correspondingly, bovine adrenal cortex cells express TRP4 abundantly. The only other TRP transcript found at considerable levels was TRP1, whereas TRP2, TRP3, TRP5(CCE2), and TRP6 were not detectable. Depletion of calcium stores with inositol 1,4,5-trisphosphate or thapsigargin activates store-operated ion channels in adrenal cells. These channels closely resemble calcium release activated Ca(2+) (CRAC) channels. Expression of trp4(CCE1) cDNA in antisense orientation significantly reduces both, the endogenous CRAC-like currents and the amount of native TRP4 protein. These results demonstrate that TRP4 contributes essentially to the formation of native CRAC-like channels in adrenal cells. PMID- 10816591 TI - Role of the C terminus of the alpha 1C (CaV1.2) subunit in membrane targeting of cardiac L-type calcium channels. AB - We have previously demonstrated that formation of a complex between L-type calcium (Ca(2+)) channel alpha(1C) (Ca(V)1.2) and beta subunits was necessary to target the channels to the plasma membrane when expressed in tsA201 cells. In the present study, we identified a region in the C terminus of the alpha(1C) subunit that was required for membrane targeting. Using a series of C-terminal deletion mutants of the alpha(1C) subunit, a domain consisting of amino acid residues 1623 1666 ("targeting domain") in the C terminus of the alpha(1C) subunit has been identified to be important for correct targeting of L-type Ca(2+) channel complexes to the plasma membrane. Although cells expressing the wild-type alpha(1C) and beta(2a) subunits exhibited punctate clusters of channel complexes along the plasma membrane with little intracellular staining, co-expression of deletion mutants of the alpha(1C) subunit that lack the targeting domain with the beta(2a) subunit resulted in an intracellular localization of the channels. In addition, three other regions in the C terminus of the alpha(1C) subunit that were downstream of residues 1623-1666 were found to contribute to membrane targeting of the L-type channels. Deletion of these domains in the alpha(1C) subunit resulted in a reduction of plasma membrane-localized channels, and a concomitant increase in channels localized intracellularly. Taken together, these results have demonstrated that a targeting domain in the C terminus of the alpha(1C) subunit was required for proper plasma membrane localization of the L type Ca(2+) channels. PMID- 10816592 TI - Tissue transglutaminase, coagulation factor XIII, and the pro-polypeptide of von Willebrand factor are all ligands for the integrins alpha 9beta 1 and alpha 4beta 1. AB - We previously reported that MOLT-3 human lymphocyte-like leukemia cells adhere to tissue-type transglutaminase (tTG) through the integrin alpha(4)beta(1). We now report that G-361 human melanoma cells also adhere to tTG, although they do not express alpha(4)beta(1). G-361 cells utilize two additional integrins, alpha(9)beta(1) and alpha(5)beta(1) to adhere to tTG. Furthermore, blood coagulation factor XIII (FXIII), another member of the transglutaminase family that is highly homologous to tTG, and propolypeptide of von Willebrand factor (pp vWF) also promoted cell adhesion through alpha(9)beta(1) or alpha(4)beta(1) in G 361 or MOLT-3 cells, respectively. In the case of pp-vWF, alpha(9)beta(1) and alpha(4)beta(1) both bind to the same site, comprised of 15 amino acid residues and designated T2-15. Moreover, SW480 human colon cancer cells stably transfected to express alpha(9)beta(1), but not mock transfectants, adhered to tTG, FXIII, pp vWF, and T2-15/bovine serum albumin conjugate. These data identify tTG, FXIII, and pp-vWF as shared ligands for the integrins alpha(9)beta(1) and alpha(4)beta(1). This report is the first to unambiguously show that these two integrins share the same cell adhesion site within one protein and provides strong support for classifying alpha(9)beta(1-) and alpha(4)-integrins as functionally related members of an integrin subfamily. PMID- 10816593 TI - alpha B-crystallin gene induction and phosphorylation by MKK6-activated p38. A potential role for alpha B-crystallin as a target of the p38 branch of the cardiac stress response. AB - The MAPK kinase MKK6 selectively stimulates p38 MAPK and confers protection against stress-induced apoptosis in cardiac myocytes. However, the events lying downstream of p38 that mediate this protection are unknown. The small heat shock protein, alphaB-crystallin, which is expressed in only a few cell types, including cardiac myocytes, may participate in MKK6-mediated cytoprotection. In the present study, we showed that, in cultured cardiac myocytes, expression of MKK6(Glu), an active form of MKK6, led to p38-dependent increases in alphaB crystallin mRNA, protein, and transcription. MKK6(Glu) also induced p38-dependent activation of the downstream MAPK-activated protein kinase, MAPKAP-K2, and the phosphorylation of alphaB-crystallin on serine-59. Initially, exposure of cells to the hyperosmotic stressor, sorbitol, stimulated MKK6, p38, and MAPKAP-K2 and increased phosphorylation of alphaB-crystallin on serine 59. However, after longer times of exposure to sorbitol, the cells began to undergo apoptosis. This sorbitol-induced apoptosis was increased when p38 was inhibited in a manner that would block alphaB-crystallin induction and phosphorylation. Thus, under these conditions, the activation of MKK6, p38, and MAPKAP-K2 by sorbitol can provide a degree of protection against stress-induced apoptosis. Supporting this view was the finding that sorbitol-induced apoptosis was nearly completely blocked in cells expressing MKK6(Glu). Therefore, the cytoprotective effects of MKK6 in cardiac myocytes are due, in part, to phosphorylation of alphaB-crystallin on serine 59 and to the induction of alphaB-crystallin gene expression. PMID- 10816594 TI - The thylakoid delta pH-dependent pathway machinery facilitates RR-independent N tail protein integration. AB - The thylakoidal DeltapH-dependent and bacterial twin arginine transport systems are structurally and functionally related protein export machineries. These recently discovered systems have been shown to transport folded proteins but are not known to assemble integral membrane proteins. We determined the translocation pathway of a thylakoidal FtsH homologue, plastid fusion/protein translocation factor, which is synthesized with a chloroplast-targeting peptide, a hydrophobic signal peptide, and a hydrophobic membrane anchor. The twin arginine motif in its signal peptide and its sole integration requirement of a DeltapH suggested that plastid fusion/protein translocation factor employs the DeltapH pathway. Surprisingly, changing the twin arginine to twin lysine or deleting the signal peptide did not abrogate integration capability or characteristics. Nevertheless, three criteria argue that all three forms require the DeltapH pathway for integration. First, integration was competed by an authentic DeltapH pathway precursor. Second, antibodies to DeltapH pathway component Hcf106 specifically inhibited integration. Finally, chloroplasts from the hcf106 null mutant were unable to integrate Pftf into their thylakoids. Thus, DeltapH pathway machinery facilitates both signal peptide-directed and N-tail-mediated membrane integration and does not strictly require the twin arginine motif. PMID- 10816595 TI - Plasma hyaluronan-binding protein is a serine protease. AB - CTCF is an essential factor for optimal transcription from the amyloid beta protein precursor promoter. A proteolytic activity detected in bovine, rabbit, horse, and human serum cleaves CTCF at three major sites, resulting in a modified mobility shift pattern of the fragments that retain DNA binding ability. The protease was purified to electrophoretic homogeneity, partially sequenced, and identified as the plasma hyaluronan-binding protein. The proteolytic activity was selectively abolished by various serine protease inhibitors, including the Kunitz type protease inhibitor domain of amyloid beta-protein precursor. Reduction with beta-mercaptoethanol showed that the 70-kDa protein consists of two polypeptides with apparent molecular masses of 44 and 30 kDa. The serine protease domain was localized to the 30-kDa polypeptide as determined by [(3)H]diisopropylfluorophosphate binding. PMID- 10816596 TI - Selectively desulfated heparin inhibits fibroblast growth factor-induced mitogenicity and angiogenesis. AB - Fibroblast growth factors (FGFs) are known to induce formation of new blood vessels, angiogenesis. We show that FGF-induced angiogenesis can be modulated using selectively desulfated heparin. Chinese hamster ovary cells (CHO677) deficient in heparan sulfate biosynthesis were employed to assess the function of heparin/heparan sulfate in FGF receptor-1 (FGFR-1) signal transduction and biological responses. In the presence of FGF-2, FGFR-1 kinase and subsequent mitogen-activated protein kinase Erk2 activities were augmented in a dose dependent manner, whereas high concentrations of heparin resulted in decreased activity. The length of the heparin oligomer, minimally an 8/10-mer, was critical for the ability to enhance FGFR-1 kinase activity. The N- and 2-O-sulfate groups of heparin were essential for binding to FGF-2, whereas stimulation of FGFR-1 and Erk2 kinases by FGF-2 also required the presence of 6-O-sulfate groups. Sulfation at 2-O- and 6-O-positions was moreover a prerequisite for binding of heparin to a lysine-rich peptide corresponding to amino acids 160-177 in the extracellular domain of FGFR-1. Selectively 6-O-desulfated heparin, which binds to FGF-2 but fails to bind the receptor, decreased FGF-2-induced proliferation of CHO677 cells, presumably by displacing intact heparin. Furthermore, FGF-2-induced angiogenesis in chick embryos was inhibited by 6-O-desulfated heparin. Thus, formation of a ternary complex of FGF-2, heparin, and FGFR-1 appears critical for the activation of FGFR-1 kinase and downstream signal transduction. Preventing complex formation by modified heparin preparations may allow regulation of FGF-2 functions, such as induction of angiogenesis. PMID- 10816597 TI - Regulation of multidrug resistance 1 (MDR1)/P-glycoprotein gene expression and activity by heat-shock transcription factor 1 (HSF1). AB - Infection of HeLa cells with adenovirus-carrying HSF1(+) cDNA, which encodes a mutated form of HSF1 with constitutive transactivation capacity, increased multidrug resistance 1 (MDR1) mRNA level and P-glycoprotein (P-gp) cell surface content and stimulated rhodamine 123 accumulation and vinblastine efflux activity. On the other hand, infection with adenovirus-carrying HSP70 and HSP27 cDNAs did not increase MDR1/P-gp expression. HSF1 regulates MDR1/P-gp expression at the transcriptional level, since HSF1(+) bound the heat-shock consensus elements (HSEs) in the MDR1 gene promoter and also activated the expression of an MDR1 promoter-driven reporter plasmid (pMDR1(-1202)). In addition, heat-shock increased pMDR1(-1202) promoter activity but not the activity of a similar reporter plasmid with point mutations at specific HSEs, and the heat-induced increase was totally inhibited by co-transfection with an expression plasmid carrying HSF1(-), a dominant negative mutant of HSF1. The stress inducers arsenite, butyrate, and etoposide also increased pMDR1(-1202) promoter activity, but the increase was not inhibited (in the case of butyrate) or was only partially inhibited (in the case of arsenite and etoposide) by HSF1(-). These results demonstrate that HSF1 regulates MDR1 expression, and that the HSEs present in the -315 to -285 region mediate the heat-induced activation of the MDR1 promoter. However, other factors may also participate in MDR1 induction by stressing agents. PMID- 10816598 TI - v-Src induces tyrosine phosphorylation of focal adhesion kinase independently of tyrosine 397 and formation of a complex with Src. AB - The non-receptor tyrosine kinase FAK plays a key role at sites of cellular adhesion. It is subject to regulatory tyrosine phosphorylation in response to a variety of stimuli, including integrin engagement after attachment to extracellular matrix, oncogene activation, and growth factor stimulation. Here we use an antibody that specifically recognizes the phosphorylated form of the putative FAK autophosphorylation site, Tyr(397). We demonstrate that FAK phosphorylation induced by integrins during focal adhesion assembly differs from that induced by activation of a temperature-sensitive v-Src, which is associated with focal adhesion turnover and transformation. Specifically, although v-Src induces tyrosine phosphorylation of FAK, there is no detectable phosphorylation of Tyr(397). Moreover, activation of v-Src results in a net decrease in fibronectin-stimulated phosphorylation of Tyr(397), suggesting possible antagonism between v-Src and integrin-induced phosphorylation. Our mutational analysis further indicates that the binding of v-Src to Tyr(397) of FAK in its phosphorylated form, which is normally mediated, at least in part, by the SH2 domain of Src, is not essential for v-Src-induced cell transformation. We conclude that different stimuli can induce phosphorylation of FAK on distinct tyrosine residues, linking specific phosphorylation events to ensuing biological responses. PMID- 10816599 TI - Functional regulation of gamma-aminobutyric acid transporters by direct tyrosine phosphorylation. AB - Tyrosine phosphorylation regulates multiple cell signaling pathways and functionally modulates a number of ion channels and receptors. Neurotransmitter transporters, which act to clear transmitter from the synaptic cleft, are regulated by multiple second messenger pathways that exert their effects, at least in part, by causing a redistribution of the transporter protein to or from the cell surface. To test the hypothesis that tyrosine phosphorylation affects transporter function and to determine its mechanism of action, we examined the regulation of the rat brain gamma-aminobutyric acid (GABA) transporter GAT1 expressed endogenously in hippocampal neurons and expressed heterologously in Chinese hamster ovary cells. Inhibitors of tyrosine kinases decreased GABA uptake; inhibitors of tyrosine phosphatases increased GABA uptake. The decrease in uptake seen with tyrosine kinase inhibitors was correlated with a decrease in tyrosine phosphorylation of GAT1 and resulted in a redistribution of the transporter from the cell surface to intracellular locations. A mutant GAT1 construct that was refractory to tyrosine phosphorylation could not be regulated by tyrosine kinase inhibitors. Activators of protein kinase C, which are known to cause a redistribution of GAT1 from the cell surface, were additive to the effects of tyrosine kinase inhibitors suggesting that multiple signaling pathways control transporter redistribution. Application of brain-derived neurotrophic factor, which activates receptor tyrosine kinases, up-regulated GAT1 function suggesting one potential trigger for the cellular regulation of GAT1 signaling by tyrosine phosphorylation. These data support the hypothesis that transporter expression and function is controlled by the interplay of multiple cell signaling cascades. PMID- 10816600 TI - Purine phosphoribosyltransferases. PMID- 10816601 TI - Metallochaperones, an intracellular shuttle service for metal ions. PMID- 10816602 TI - Transcriptional control in myelinating glia: flavors and spices. AB - Transcriptional control in myelinating glia is often described in terms of a handful of trans-acting proteins with preferential expression in these cells. An equally valid approach is the identification of cis-acting elements in genes, which are specifically transcribed in myelinating glia. Regulatory regions of several myelin genes have been analyzed in transgenic animals, transient transfections and in vitro. In some cases, these studies have identified regions responsible for glial expression within the promoters or immediate upstream regions. Other myelin genes possess promoters, which simply secure basal levels of transcription, but do not contain glia-specific cis-acting elements. Promoters of myelin genes also differ strongly in other respects. They either contain a TATA-box or are TATA-less and GC-rich. They exhibit multiple transcription initiation sites or a single strong one. Binding sites for general transcription factors, such as NF-I, Sp1, and CAAT-box binding proteins, and for downstream effectors of major signaling pathways are found in them in abundance. In agreement, members of the AP-1, CREB, STAT, and NF-kappaB families are well described components of the transcription machinery in myelinating glia. Together with several members of the nuclear receptor family, they are an intrinsic part of the transcriptional control in myelinating glia. PMID- 10816604 TI - Aquaporin-4-containing astrocytes sustain a temperature- and mercury-insensitive swelling in vitro. AB - In order to understand the molecular mechanism underlying astroglial swelling, we studied primary astrocyte cultures from newborn mouse and analyzed them for expression of functional water channels. Immunocytochemical analysis of mouse brain confirms the presence of AQP4 location in astrocytic endfeet with a polarized pattern, as found in rat. Using Southern blot PCR and Western blot analysis, we demonstrate that primary astrocyte cultures from mouse express the AQP4 water channel at both the RNA and protein levels. Two polypeptides, of 30 kDa and 32 kDa, were identified in the astrocytes. Densitometric analysis demonstrates that the 32-kDa form represents 25% of the total AQP4 protein. Moreover, immunofluorescence experiments show strong surface membrane expression of AQP4 protein in cultured cells, even though the polarity of the expression is not maintained. Furthermore, functional studies indicate that cultured astrocytes manifest rapid and temperature-independent volume changes in response to osmotic gradients, in agreement with a channel-mediated water transport. Water movement was found to be HgCl(2) insensitive, suggesting AQP4 and AQP7 as putative water channels. Using Western blot and PCR experiments, we exclude the presence of AQP7 in astrocytes, indicating that only AQP4 is responsible for the rapid water movement. Altogether, the results indicate that primary astrocyte cultures are a valid cell model for further investigation of the molecular mechanism of water movement in the brain and its physiological regulation. PMID- 10816603 TI - Unloading and refilling of two classes of spatially resolved endoplasmic reticulum Ca(2+) stores in astrocytes. AB - Signaling by two classes of endoplasmic reticulum (ER) Ca(2+) stores was studied in primary cultured rat astrocytes. Cytosolic and intra-ER Ca(2+) concentrations ([Ca(2+)](CYT) and [Ca(2+)](ER)) were measured with, respectively, Fura-2 and Furaptra, in separate experiments. The agonists, glutamate and ATP, released Ca(2+) primarily from cyclopiazonic acid (CPA)-sensitive ER Ca(2+) stores (CPA inhibits ER Ca(2+) pumps). Agonist-evoked release was abolished by prior treatment with CPA but was unaffected by prior depletion of caffeine/ryanodine (CAF/RY)-sensitive ER Ca(2+) stores. Conversely, prior depletion of the CPA sensitive stores did not interfere with Ca(2+) release or reuptake in the CAF/RY sensitive stores. Unloading of the CPA-sensitive stores, but not the CAF/RY sensitive stores, promoted Ca(2+) entry through "store-operated channels." Resting [Ca(2+)](ER) averaged 153 microM (based on in situ calibration of Furaptra: K(D) = 76 microM, vs 53 microM in solution). The releasable Ca(2+) in both types of ER Ca(2+) stores was increased by Na(+) pump inhibition with 1 mM ouabain or K(+)-free medium. Using high spatial resolution imaging and image subtraction methods, we observed that some regions of the ER (45-58% of the total ER) unloaded and refilled when CPA was added and removed. Other regions of the ER (24-38%) unloaded and refilled when CAF was added and removed. The overlap between these two classes of ER was only 10-18%. These data indicate that there are two structurally separate, independent components of the ER and that they are responsible for the functional independence of the CPA-sensitive and CAF/RY sensitive ER Ca(2+) stores. PMID- 10816605 TI - Glial responses, clusterin, and complement in permanent focal cerebral ischemia in the mouse. AB - There is considerable evidence that complement activation occurs within the CNS in inflammatory and degenerative disorders, but little is known about its involvement in the pathophysiology of cerebral ischemia. Our study sought to characterize the glial response and the expression of complement factors after permanent focal cerebral ischemia in the mouse, using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and immunohistochemistry. mRNA expression of glial fibrillary acidic protein (GFAP) increased at day 1 and peaked 3 days after middle cerebral artery (MCA) occlusion in the perifocal area. Immunohistochemical staining for GFAP indicated that astroglia were activated the day after MCA occlusion. Microglial activation, as assessed by lectin-binding experiments, increased by 1 day after MCA occlusion in the perifocal area and peaked at 3 days postocclusion. RT-PCR experiments demonstrated an increased expression of clusterin, C1qB, and C4 mRNA in the ischemic cortex, with a peak level at 3 days after MCA occlusion. Clusterin, C1qB, and C4 mRNA were located in the perifocal area, as assessed by in situ hybridization. Reactive astrocytes within the cortex medial to the ischemic lesion were found to be strongly immunoreactive for clusterin. In addition, we observed C1q-positive macrophage-like cells within the infarcted core at 3 days postocclusion. At 7 days after the onset of ischemia, increased C4 immunostaining was restricted to perifocal neurons. We conclude that local expression of complement components may contribute to the inflammation observed in this model, thereby representing an important process in secondary injury mechanisms after focal cerebral ischemia. PMID- 10816606 TI - Adenosine-induced expression of interleukin-6 in astrocytes through protein kinase A and NF-IL-6. AB - In various neurologic diseases, astrocytes express interleukin-6 (IL-6), which is an endogenous pyrogen, a neuroprotective factor, and a regulator of the blood brain barrier. The expression of IL-6 in astrocytes is stimulated by extracellular adenosine through A(2B) receptors. To investigate the signaling cascade that induces IL-6 gene transcription further, we transfected primary mouse astrocytes with a reporter gene construct, in which luciferase expression is directed by the human IL-6 promoter. Expression of PKI, an inhibitor of protein kinase A (PKA), interfered with IL-6 transcription indicating that PKA mediates the effect of adenosine. The CAAT box of the IL-6 promoter is necessary for the stimulation by adenosine as a mutation in this element reduced the stimulation by adenosine. Indeed, the cAMP agonist forskolin increased the binding of the transcription factors NF-IL-6 and C/EBPdelta to the CAAT box of the IL-6 promoter in nuclear extracts of astrocytes. Inhibition of the de novo synthesis of NF-IL-6 by cycloheximide or an antisense oligonucleotide reduced the enhancement of NF-IL-6 binding to the CAAT box and inhibited stimulation of IL-6 transcription by forskolin. In addition, overexpression of NF-IL-6 induced IL-6 transcription. This suggests that adenosine induces the de novo synthesis of NF IL-6 through activation of PKA and thereby stimulates transcription of IL-6 in astrocytes. PMID- 10816607 TI - Rat oligodendrocytes express the vitamin D(3) receptor and respond to 1,25 dihydroxyvitamin D(3). AB - The present study investigates the presence of vitamin D receptor (VDR) in cells of the rat oligodendrocyte (OL) lineage. VDR transcripts were detected by in situ hybridization in a fraction of rat OL in secondary cultures. The VDR protein was shown to be co-localized in cells that are also recognized by an anti-myelin basic protein (MBP) antibody. Likewise, in vivo, VDR-positive cells were found in the brain white matter, such as the internal capsule of the striatum or the corpus callosum but also in the spinal cord. At least part of these positive cells in vivo correspond to OL, since they were co-stained by an anti-carbonic anhydrase II antiserum. Northern blot analyses of the CG-4 OL cell line demonstrated that the VDR transcripts are already found in the O-2A precursors. There was a two-fold increase in the relative abundance of these transcripts in differentiated OL or in type-2 astrocytes. 1, 25-dihydroxyvitamin D(3) [1,25 (OH)(2)D(3)] increased the pool of transcripts encoding its own receptor, the VDR. The hormone also enhanced the abundance of the mRNA of the nerve growth factor (NGF) and of its low-affinity receptor, the p75(NTR) protein. By contrast, the hormone had no effect on the levels of MBP or proteolipid protein (PLP) mRNA. This finding suggests that unlike retinoic acid (RA) or thyroid hormone, 1,25 (OH)(2)D(3) has no regulatory action on the synthesis of myelin proteins. PMID- 10816608 TI - Comparative anatomy of the cerebellar cortex in mice lacking vimentin, GFAP, and both vimentin and GFAP. AB - In the cerebellum of adult mammals, glial fibrillary acidic protein (GFAP) and vimentin (VIM) are coexpressed in Golgi epithelial cells (GEC), also known as Bergmann glia. In this study we used three transgenic knockout mice (GFAP, VIM and double GFAP and VIM) to analyze the involvement of these proteins in the building of glial filaments and in neuron-glia interactions. The cerebella of VIM, GFAP, and GFAP/VIM mutant mice were processed by the rapid Golgi method and also for electron microscopy. In VIM mutant mice, Bergmann fibers are hypertrophic with thickened appendages. In the electron microscope they appear as large glial profiles devoid of glial filaments, with embedded dendritic thorns and parallel fiber boutons. In addition, signs of degeneration are observed in Purkinje cells. In GFAP mutant mice, GEC exhibit fine, delicate processes, as those seen in wild-type animals, however, a large accumulation of lamellae and granular appendages was observed along their surfaces, which came into contact with each other. The electron microscope exhibited fine and scarce astroglial profiles containing some glial filaments, a stunted glia limitans, and the presence of large extracellular spaces. In double mutant mice, the two phenotypes are expressed but appear attenuated, with a total absence of glial filaments and the general appearance of immaturity for GEC. In conclusion, it appears that the absence of each of the proteins yields a specific phenotype and that the defects are not necessarily additive. PMID- 10816609 TI - Spatial distribution of spermine/spermidine content and K(+)-current rectification in frog retinal glial (Muller) cells. AB - Previous studies in retinal glial (Muller) cells have suggested that (1) the dominant membrane currents are mediated by K(+) inward-rectifier (Kir) channels (Newman and Reichenbach, Trends Neurosci 19:307-312, 1996), and (2) rectification of these Kir channels is due largely to a block of outward currents by endogenous polyamines such as spermine/spermidine (SPM/SPD) (Lopatin et al., Nature 372:366 369, 1994). In frog Muller cells, the degree of rectification of Kir-mediated currents is significantly higher in the endfoot than in the somatic membrane (Skatchkov et al., Glia 27:171-181, 1999). This article shows that in these cells there is a topographical correlation between the local cytoplasmic SPM/SPD immunoreactivity and the ratio of inward to outward K(+) currents through the surrounding membrane area. Throughout the retina, Muller cell endfeet display a high SPM/SPD immunolabel (assessed by densitometry) and a large inward rectification of K(+) currents, as measured by the ratio of inward to outward current produced by step changes in [K(+)](o). In the retinal periphery, Muller cell somata are characterized by roughly one-half of the SPM/SPD immunoreactivity and K(+)-current rectification as the corresponding endfeet. In the retinal center, Muller cell somata are virtually devoid of both SPM/SPD immunolabel and K(+)-current inward rectification. Comparing one region of the retina with another, we find an exponential correlation between the local K(+) rectification and the local SPM/SPD content. This finding suggests that the degree of inward rectification in a given membrane area is determined by the local cytoplasmic polyamine concentration. PMID- 10816610 TI - Transient hypoxia/hypoglycemia upregulates endothelin B receptors in cultured rat astrocytes. AB - Endothelins are potent vasoactive peptides that bind to their specific receptors, playing an important role in the CNS under physiological and pathophysiological conditions. Astrocytes, which have been shown to express these receptors, also have a considerable role to play under physiological and pathophysiological conditions, particularly those involved in delayed neuronal death. We carried out in vitro receptor autoradiographic binding experiments using specific ligands for endothelin receptors on cultured rat astrocytes. On astrocytes, the specific binding sites for (125)I-PD151242 (a selective endothelin A receptor antagonist) and (125)I-IRL 1620 (a selective endothelin B receptor agonist) were detected. We also characterized the qualitative and quantitative changes of endothelin receptors 24 h after subjecting cultured rat astrocytes to a transient 4-h hypoxia/hypoglycemia insult, used as a model of delayed neuronal death. After transient hypoxia/hypoglycemia, the number of endothelin B receptors increased significantly on cultured astrocytes, but this did not occur among the endothelin A receptors. These findings suggest that the astrocytic effects associated with endothelin in delayed neuronal death include gliosis or the repair process or both, manifested primarily by an increase in the number of endothelin B receptors. This rise does not require interaction with other types of CNS cells. Endothelin A receptors might have a role taking their number into consideration on rat astrocytes under both physiological and pathophysiological conditions. PMID- 10816611 TI - A clinical and bacteriological study of children suffering from haemolytic uraemic syndrome in Tucuman, Argentina. AB - Haemolytic uraemic syndrome (HUS) is a disease with serious consequences for children, such as terminal chronic renal failure. During the last few years there have been numerous studies undertaken to determine whether there is a relationship between this disease and the presence of Shiga toxin-producing bacteria. Escherichia coli (E. coli) O157:H7 is one of the most frequent etiologic agents of HUS. It acts through cytotoxins called Shiga toxin 1 (Stx1) and/or Shiga toxin 2 (Stx2) and carries a 90-Kb plasmid codified for an adhesion fimbria which is part of its pathogenicity. The objectives of this study were to: 1). confirm whether there exists a relationship between severity and clinical presentation of HUS; 2). prove the existence of Stx1 and/or Stx2 in the faeces of HUS patients; and 3). detect the presence of Stx1- and/or Stx2-producing E. coli. Our results did not show any difference in the average age, sex or clinical behavior between children with diarrhea positive (D+) HUS and diarrhea negative (D-) HUS. Male patients were predominant, as was incidence during summer, considering all cases. Nor could we find any relationship between severity and HUS type. E. coli O157:H7 was isolated in 40% of the patients with (D+) HUS and in 50% of patients with (D-) HUS. Another serotype, O55:K59, was also isolated (7%). Stx1 and/or Stx2 were found in all HUS cases. The following virulence factors of E. coli strains isolated from 12 patients were found: Adhesion fimbria (100%), Stx1 (16%), Stx2 (32%), and Stx1 + Stx2 (50%). None of these factors was found in control patients. Sixty-three percent of the HUS cases showed seroconversion for lipopolysaccharides of E. coli O157. We drew the following conclusions: 1). there is no significant relationship between seriousness of HUS and type of disease; 2). an association exists between HUS and the production of Stx1 and Stx2; 3). the incidence of E. coli O157:H7 was high in Tucuman, Argentina; and 4). Stx2 alone or in association with Stx1 was the predominant toxin. PMID- 10816612 TI - Molecular typing of vibrio vulnificus isolated from clinical specimens by pulsed field gel electrophoresis and random amplified polymorphic DNA analysis. AB - Primary V. vulnificus septicemia has been continuously reported in Korea. We analyzed the molecular diversity of V. vulnificus strains isolated from clinical specimens using pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) analysis. We analyzed a total of 23 V. vulnificus strains isolated from 22 patients between 1992 and 1997. RAPD analysis was performed using five random primers, and we obtained chromosomal DNA restriction patterns with NotI and SfiI by PFGE. Two isolates from one patient disclosed similarity value 1.0 by RAPD and had an indistinguishable pattern when analyzed with PFGE, which indicated that they were the same strains. The remaining 22 isolates from 22 patients could be separated into 5 different molecular types in RAPD analysis. The range of similarity values among the isolates was wide (0.29-0.81). Of 22 strains, four strains could not be typed in repeated trials by PFGE with NotI and SfiI. The PFGE patterns of 18 strains showed a remarkable polymorphism consisting 12-19 fragments (20-870 kb). These results show that V. vulnificus strains isolated from Korea are genetically diversified for the most part. PMID- 10816613 TI - Skin reaction to yellow fever vaccine after immunization with rabies vaccine of chick embryo cell culture origin. AB - Skin reaction to yellow fever vaccine was examined after immunization with rabies vaccine. The two vaccines contained substrates from chick embryo cells (rabies vaccine) and chick whole embryo (yellow fever attenuated vaccine), as well as gelatin. A prick test with gelatin showed negative results in all vaccinees examined. An intradermal skin test revealed that the yellow fever vaccine had reacted with an anti-egg protein antibody-like substance in a case with a history of egg allergy before rabies vaccination. A case inoculated two times with the rabies vaccine revealed a positive reaction to egg-white protein as well as the yellow fever vaccine. This case had no anamnesis of egg allergy. Thus, an antibody reactive to the egg-white protein and/or the yellow fever vaccine was inducible by the rabies vaccine. The reaction of this antibody was not systemic but local at the skin test by the yellow fever vaccine. The period of the rabies vaccine sensitization reactive to the yellow fever vaccine could be estimated as longer than 14.3 +/- 9.6 days (mean +/- SD), based on a follow-up examination of the positive skin reaction in 41 of 84 cases examined. We therefore conclude that the yellow fever vaccine can be safely administered at an interval of at least four weeks after a second rabies vaccination. PMID- 10816614 TI - Il-1, EGF, and HGF suppress the antiviral activity of interferon in primary monkey hepatic parenchymal cells. AB - The purpose of this study was to elucidate the mechanism behind low efficacy of interferon therapy to hepatitis C virus infected patients by using primary monkey hepatic parenchymal cells as a surrogate of primary human liver cells. The effects of various cytokines on the antiviral activity of IFNs in the monkey hepatic cells were studied to search for physiological inhibitors. Interleukin-1 alpha, EGF, and HGF showed suppressive effects on the antiviral activity of IFN alpha, -beta in primary monkey hepatic cells when examined by the yield reduction method using vesicular stomatitis virus (VSV). In contrast, 50 ng/ml of TNF and IL-6 had no suppressive effect on the IFN-induced antiviral state in the hepatic cells. PMID- 10816615 TI - Paragonimiasis acquired by eating boar meat: current status in Japan. PMID- 10816616 TI - Detection of adeno-associated virus type 2 in patients with viral infection. PMID- 10816617 TI - Detection of a multi-prefectural E coli O157:H7 outbreak caused by contaminated Ikura-Sushi ingestion. PMID- 10816618 TI - Outbreak of salmonellosis caused by ingestion of cuttlefish chips: epidemiological analysis by pulsed-field gel electrophoresis. PMID- 10816619 TI - Annual Surveillance Report of HIV/AID in Japan, 1997. AIDS Surveillance Committee, Ministry of Health and Welfare, Japan. Working Group of Annual AIDS Surveillance, Ministry of Health and Welfare, Japan. PMID- 10816620 TI - Carotid endarterectomy in the community hospital in patients age 80 and older. AB - OBJECTIVE: To determine whether the rates of death and complications of carotid endarterectomy (CE) were different in the octogenarian population than in patients younger than age 80. SUMMARY BACKGROUND DATA: The utility of CE depends on the ability of the surgeon and hospital to attain low rates of death and complications, including all subgroups of the patient population. In the past 30 years, the number of people age 85 and older has increased 274%. METHODS: Detailed chart review was carried out on all CE procedures done from 1979 through 1998. Descriptive demographic data, risk factors, surgical details, length of stay, deaths, and complications were recorded. RESULTS: A total of 2,398 CEs were performed in 1,970 patients; 2,180 procedures were performed in 1,783 patients younger than 80, and 218 CEs were performed in 187 patients age 80 and older. Sixty-five percent of the octogenarians and 67% of patients younger than age 80 had neurologic symptoms. Among asymptomatic patients, 89% had stenosis of 75% or more. There were 62 strokes in the 2,180 procedures in the younger group, for a stroke rate of 2.8%, and 7 strokes in the 218 procedures in the older group, for a stroke rate of 3.2%. The death rates were 0.9% for the octogenarians and 1.4% for the younger group. CONCLUSIONS: Carotid endarterectomy can be safely performed in a community hospital in patients age 80 and older. Outcomes in octogenarians were not significantly different than those of younger patients and were within the range required for CE to be considered beneficial in the prevention of stroke. PMID- 10816621 TI - Treatment of experimentally induced caval thrombosis with oral low molecular weight heparin and delivery agent in a porcine model of deep venous thrombosis. AB - OBJECTIVE: This experiment evaluated enterally administered low molecular weight heparin (LMWH) combined with sodium N-[10-(2-hydroxybenzoyl)amino] decanoate (SNAD) for the treatment of induced venous thrombosis. SUMMARY BACKGROUND DATA: SNAD is a delivery agent that potentiates the gastrointestinal absorption of LMWH. METHODS: Forty female pigs were equally assigned to four groups: control (saline); enteral LMWH, 2,000 IU/kg; enteral SNAD, 50 mg/kg; and enteral LMWH, 2,000 IU/kg and SNAD, 50 mg/kg. Under fluoroscopic guidance, the infrarenal vena cava was occluded with a balloon catheter. Two milliliters of ethanol was injected into the distal vena cava. The inflated balloon catheter remained in situ for 5 days, at which time animals angiographically exhibiting thrombus were randomly assigned to the four groups. Study medications were dosed at 12-hour intervals by means of a gastrostomy tube placed previously. After 7 days of treatment, thrombus was extracted. A separate group of 10 animals was used to measure plasma antifactor Xa levels for 6 hours after enteral dosing of LMWH/SNAD. RESULTS: The amount of residual thrombus after treatment with enteral LMWH/SNAD was significantly decreased. Antifactor Xa levels were significantly elevated in the LMWH/SNAD group versus baseline. CONCLUSION: The combination of enterally administered LMWH and SNAD given for 7 days appeared to decrease caval thrombosis in this model of deep vein thrombosis. Enteral LMWH/SNAD effected an increase in plasma levels of antifactor Xa. PMID- 10816622 TI - Clinical relevance of molecular staging for melanoma: comparison of RT-PCR and immunohistochemistry staining in sentinel lymph nodes of patients with melanoma. AB - OBJECTIVE: To determine the clinical significance of a molecular assay based on the reverse transcriptase polymerase chain reaction (RT-PCR) for the presence of micrometastatic melanoma cells in sentinel lymph nodes (SLNs). SUMMARY BACKGROUND DATA: Routine histologic examination of lymph nodes often underestimates the presence of micrometastatic disease. The authors have previously shown that an RT PCR assay designed to detect melanocyte-specific expression of the tyrosinase gene could be used to define a population of patients at higher risk for both recurrence and death compared with routine hematoxylin and eosin (H&E) histology. In this study, the authors used the tyrosinase RT-PCR assay in a patient population examined by a more detailed histologic analysis, including S-100 immunohistochemistry. METHODS: Patients underwent lymphatic mapping and SLN biopsy. SLN specimens were bivalved, and half of each specimen was serially sectioned and examined by routine H&E histology and S-100 immunohistochemistry. The other half of each specimen was analyzed by a nested RT-PCR assay. RESULTS: Hematoxylin and eosin histology detected metastatic disease in 36 (16%) of the 233 patients tested. S-100 immunohistochemistry detected micrometastatic disease in another 16 patients, and 114 (63%) of 181 patients with histology-negative nodes had positive findings on RT-PCR. There were significant differences between PCR-positive and PCR-negative patient groups in Breslow thickness, Clark level, and the presence of ulceration of the primary tumor, factors that have been shown to correlate with recurrence and survival. CONCLUSIONS: These results suggest that RT-PCR can increase the sensitivity of detection of metastatic melanoma cells in SLNs over the current standard methods, including H&E histology and S 100 immunohistochemistry. Further long-term follow-up is needed to detect actual differences in recurrence and overall survival. PMID- 10816623 TI - Blunt hepatic injury: a paradigm shift from operative to nonoperative management in the 1990s. AB - OBJECTIVE: To analyze the outcome of hemodynamically stable patients with blunt hepatic injury managed nonoperatively, and to examine the impact of this approach on the outcome of all patients with blunt hepatic injury. SUMMARY BACKGROUND DATA: Until recently, operative management has been the standard for liver injury. A prospective trial from the authors' institution had shown that nonoperative management could safely be applied to hemodynamically stable patients with blunt hepatic injury. The present study reviewed the authors' institutional experience with blunt hepatic trauma since that trial and compared the results with prior institutional experience. METHODS: Six hundred sixty-one patients with blunt hepatic trauma during the 5-year period ending December 1998 were reviewed (NONOP2). The outcomes were compared with two previous studies from this institution: operative 1985 to 1990 (OP) and nonoperative 1993 to 1994 (NONOP1). RESULTS: All 168 OP patients were managed operatively. Twenty-four (18%) of 136 NONOP1 patients and 101 (15%) of the 661 NONOP2 patients required immediate exploration for hemodynamic instability. Forty-two (7%) patients failed nonoperative management; 20 were liver-related. Liver-related failures of nonoperative management were associated with higher-grade injuries and with larger amounts of hemoperitoneum on computed tomography scanning. Twenty-four hour transfusions, abdominal infections, and hospital length of stay were all significantly lower in the NONOP1 and NONOP2 groups versus the OP cohort. The liver-related death rate was constant at 4% in the three cohorts over the three time periods. CONCLUSIONS: Although urgent surgery continues to be the standard for hemodynamically compromised patients with blunt hepatic trauma, there has been a paradigm shift in the management of hemodynamically stable patients. Approximately 85% of all patients with blunt hepatic trauma are stable. In this group, nonoperative management significantly improves outcomes over operative management in terms of decreased abdominal infections, decreased transfusions, and decreased lengths of hospital stay. PMID- 10816624 TI - Choice of surgical technique influences perioperative outcomes in liver transplantation. AB - OBJECTIVE: To examine how the choice of surgical technique influenced perioperative outcomes in liver transplantation. SUMMARY BACKGROUND DATA: The standard technique of orthotopic liver transplantation with venovenous bypass (VVB) is commonly used to facilitate hemodynamic stability. However, this traditional procedure is associated with unique complications that can be avoided by using the technique of liver resection without caval excision (the piggyback technique). METHODS: A prospective comparison of the two procedures was conducted in 90 patients (34 piggyback and 56 with VVB) during a 2.5-year period. Although both groups had similar donor and recipient demographic characteristics, posttransplant outcomes were significantly better for the patients undergoing the piggyback technique. The effect of surgical technique was examined using a stepwise approach that considered its impact on two levels of perioperative and postoperative events. RESULTS: The analysis of the first level of perioperative events found that the piggyback procedure resulted in a 50% decrease in the duration of the anhepatic phase. The analysis of the second level of perioperative events found a significant relation between the anhepatic phase and the duration of surgery and between the anhepatic phase and the need for blood replacement. The analysis of the first level of postoperative events found that the intensive care unit stay was significantly related to both the duration of surgery and the need for blood replacement. The intensive care unit stay was in turn related to the second level of postoperative events, namely the length of hospital stay. Finally, total charges were directly related to length of hospital stay. The overall 1-year actuarial patient and graft survival rates were 94% in the piggyback and 96% in the VVB groups, respectively. CONCLUSIONS: These data demonstrate that surgical choices in complex procedures such as orthotopic liver transplantation trigger a chain of events that can significantly affect resource utilization. In the current healthcare climate, examination of the sequence of events that follow a specific treatment may provide a more complete framework for choosing between treatment alternatives. PMID- 10816625 TI - Surgical management of anatomical variations of the right lobe in living donor liver transplantation. AB - OBJECTIVE: To review the anatomical variations of the right lobe encountered in 40 living liver donors, describe the surgical management of these variations, and summarize the results of these procedures. SUMMARY BACKGROUND DATA: Anatomical variability is the rule rather than the exception in liver and biliary surgery. To make effective use of liver segments from living donors for transplantation, surgical techniques must be adapted to the anomalies. METHODS: Donor evaluation included celiac and mesenteric angiography with portal phase, magnetic resonance angiography, and intraoperative ultrasonography and cholangiography. Arterial anastomoses were generally between the donor right hepatic artery and the recipient main hepatic artery. Jump-grafts were constructed for recipients with hepatic artery thrombosis, and double donor arteries were joined to the bifurcation of the recipient hepatic artery. The branches of a trifurcated donor portal vein were isolated during the parenchymal transection, joined in a common cuff, and anastomosed to the recipient main portal vein. Significant accessory hepatic veins were preserved, brought together in a common cuff if multiple, and anastomosed to the recipient cava. The bile ducts were individually drained through a Roux-en-Y limb, and stents were placed in most patients. RESULTS: Forty right lobe liver transplants were performed between adults. No donor was excluded because of prohibitive anatomy. Seven recipients had a prior transplant and five had a transjugular intrahepatic portosystemic shunt (TIPS). Arterial anomalies were noted in six donors and portal anomalies in four. Arterial jump-grafts were required in three. Sixteen had at least one significant accessory hepatic vein, and one had a double right hepatic vein. There were no vascular complications. Multiple bile ducts were found in 27 donors. Biliary complications occurred in 33% of patients without stents and 4% with stents. CONCLUSIONS: Anatomical variations of the right lobe can be accommodated without donor complications or complex reconstruction. Previous transplantation and TIPS do not significantly complicate right lobe transplantation. Microvascular arterial anastomosis is not necessary, and vascular complications should be infrequent. Biliary complications can be minimized with stenting. PMID- 10816626 TI - Single-layer continuous versus two-layer interrupted intestinal anastomosis: a prospective randomized trial. AB - OBJECTIVE: To determine the suitability of a single-layer continuous technique for intestinal anastomosis in a surgical training program. SUMMARY BACKGROUND DATA: Several recent reports have advocated the use of a continuous single-layer technique for intestinal anastomosis. Purported advantages include shorter time for construction, lower cost, and perhaps a lower rate of anastomotic leakage. The authors hypothesized that the single-layer continuous anastomosis could be safely introduced into a surgical training program and that it could be performed in less time and at a lower cost than the two-layer interrupted anastomosis. METHODS: The study was conducted during a 3-year period ending September 1999. All adult patients requiring intestinal anastomosis were considered eligible. Patients who required anastomosis to the stomach, duodenum, and rectum were excluded. Patients were also excluded if the surgeon did not believe either technique could be used. Patients were randomly assigned to one- or two-layer techniques. Single-layer anastomoses were performed with a continuous 3-0 polypropylene suture. Two-layer anastomoses were constructed using interrupted 3 0 silk Lembert sutures for the outer layer and a continuous 3-0 polyglycolic acid suture for the inner layer. The time for anastomosis began with the placement of the first stitch and ended when the last stitch was cut. Anastomotic leak was defined as radiographic demonstration of a fistula or nonabsorbable material draining from a wound after oral administration, or visible disruption of the suture line during reexploration. RESULTS: Sixty-five single-layer and 67 two layer anastomoses were performed. The groups were evenly matched according to age, sex, diagnosis, and location of the anastomosis. Two leaks (3.1%) occurred in the single-layer group and one (1.5%) in the two-layer group. Two abscesses (3.0%) occurred in each group. A mean of 20.8 minutes was required to construct a single-layer anastomosis versus 30.7 minutes for the two-layer technique. Mean length of stay was 7.9 days for single-layer patients and 9.9 days for two-layer patients; this difference did not quite reach statistical significance. Cost of materials was $4.61 for the single-layer technique and $35.38 for the two-layer method. CONCLUSIONS: A single-layer continuous anastomosis can be constructed in significantly less time and with a similar rate of complications compared with the two-layer technique. It also costs less than any other method and can be incorporated into a surgical training program without a significant increase in complications. PMID- 10816627 TI - Gut peptide receptor expression in human pancreatic cancers. AB - OBJECTIVE: To determine the prevalence of gastrointestinal (GI) peptide receptor expression in pancreatic cancers, and to further assess signaling mechanisms regulating neurotensin (NT)-mediated pancreatic cancer growth. SUMMARY BACKGROUND DATA: Pancreatic cancer remains one of the leading causes of GI cancer death; novel strategies for the early detection and treatment of these cancers is required. Previously, the authors have shown that NT, an important GI hormone, stimulates the proliferation of an NT receptor (NTR)-positive pancreatic cancer. METHODS: A total of 26 human pancreatic adenocarcinomas, obtained after resection, and 5 pancreatic cancer xenografts were analyzed for expression of NTR, vasoactive intestinal peptide receptor (VIPR), substance P receptor (SPR), and gastrin-releasing peptide receptor (GRPR). In addition, NTR expression, [Ca2+]i mobilization, and growth in response to NT was determined in L3.6, a metastatic pancreatic cancer cell line. RESULTS: Neurotensin receptor was expressed in 88% of the surgical specimens examined and all five of the pancreatic cancer xenografts. In contrast, VIPR, SPR, and GRPR expression was detected in 31%, 27%, and 8% of pancreatic cancers examined, respectively. Expression of NTR, functionally coupled to the Ca2+ signaling pathway, was identified in L3.6 cells; treatment with NT (10 micromol/L) stimulated proliferation of these cells. CONCLUSIONS: The authors demonstrated NTR expression in most of the pancreatic adenocarcinomas examined. In contrast, VIPR, SPR, and GRPR expression was detected in fewer of the pancreatic cancers. The expression of NTR and other peptide receptors suggests the potential role of endocrine manipulation in the treatment of these cancers. Further, the presence of GI receptors may provide for targeted chemotherapy or radiation therapy or in vivo scintigraphy for early detection. PMID- 10816628 TI - Academic health systems management: the rationale behind capitated contracts. AB - OBJECTIVE: To determine why hospitals enter into "capitated" contracts, which often generate accounting losses. The authors' hypothesis is that hospitals coordinate contracts to keep beds full and that in principal, capitated contracts reflect sound capacity management. SUMMARY BACKGROUND DATA: In high-overhead industries, different consumers pay different prices for similar services (e.g., full-fare vs. advanced-purchase plane tickets, full tuition vs. financial aid). Some consumers gain access by paying less than total cost. Hospitals, like other high-overhead business enterprises, must optimize the use of their capacity, amortizing overhead over as many patients as possible. This necessity for enhanced throughput forces hospitals and health systems to discount empty beds, sometimes to the point where they incur accounting losses serving some payors. METHODS: The authors analyzed the cost accounting system at their university teaching hospital to compare hospital and intensive care unit (ICU) lengths of stay (LOS), variable direct costs (VDC), overhead of capitated patients, and reimbursement versus other payors for all hospital discharges (n = 29,036) in fiscal year 1998. The data were analyzed by diagnosis-related groups (DRGs), length of stay (LOS), insurance carrier, proximity to hospital, and discharge disposition. Patients were then distinguished across payor categories based on their resource utilization, proximity to the hospital, DRG, LOS, and discharge status. RESULTS: The mean cost for capitated patients was $4,887, less than half of the mean cost of $10,394 for the entire hospitalized population. The mean capitated reimbursement was $928/day, exceeding the mean daily VDC of $616 but not the total cost of $1,445/day. Moreover, the mean total cost per patient day of treating a capitated patient was $400 less than the mean total cost per day for noncapitated patients. The hospital's capitated health maintenance organization (HMO) patients made up 16. 0% of the total admissions but only 9.4% of the total patient days. Both the mean LOS of 3.4 days and the mean ICU LOS of 0.3 days were significantly different from the overall values of 5.8 days and 1 day, respectively, for the noncapitated population. For patients classified with a DRG with complication who traveled from more than 60 miles away, the mean LOS was 10.7 days and the mean total cost was $21,658. This is in contrast to all patients who traveled greater than 60 miles, who had an LOS of 7.2 days and a mean total cost of $12,569. CONCLUSION: The capitated payor directed the bulk of its subscribers to one hospital (other payors transferred their sicker patients). This was reflected in the capitated group's lower costs and LOS. This stable stream of relatively low-acuity patients enhanced capacity utilization. For capitated patients, the hospital still benefits by recovering the incremental cost (VDC) of treating these patients, and only a portion of the assigned overhead. Thus, in the short run, capitated patients provide a positive economic benefit. Other payors' higher-acuity patients arrive more randomly, place greater strains on capacity, and generate higher overhead costs. This results in differential reimbursement to cover this incremental overhead. Having a portfolio of contracts allows the hospital to optimize capacity both in terms of patient flows and acuity. One risk of operating near capacity is that capitated patients could displace other higher-paying patients. PMID- 10816629 TI - Financial impact of tertiary care in an academic medical center. AB - OBJECTIVE: To analyze the financial impact of three complex vascular surgical procedures to both an academic hospital and a department of surgery and to examine the potential impact of decreased reimbursements. SUMMARY BACKGROUND DATA: The cost of providing tertiary care has been implicated as one potential cause of the financial difficulties affecting academic medical centers. METHODS: Patients undergoing revascularization for chronic mesenteric ischemia, elective thoracoabdominal aortic aneurysm repair, and treatment of infected aortic grafts at the University of Florida were compared with those undergoing elective infrarenal aortic reconstruction and carotid endarterectomy. Hospital costs and profit summaries were obtained from the Clinical Resource Management Office. Departmental costs and profit summary were estimated based on the procedural relative value units (RVUs), the average clinical cost per RVU ($33.12), surgeon charges, and the collection rate for the vascular surgery division (30.2%) obtained from the Faculty Group Practice. Surgeon work effort was analyzed using the procedural work RVUs and the estimated total care time. The analyses were performed for all payors and the subset of Medicare patients, and the potential impact of a 15% reduction in hospital and physician reimbursement was analyzed. RESULTS: Net hospital income was positive for all but one of the tertiary care procedures, but net losses were sustained by the hospital for the mesenteric ischemia and infected aortic graft groups among the Medicare patients. In contrast, the estimated reimbursement to the department of surgery for all payors was insufficient to offset the clinical cost of providing the RVUs for all procedures, and the estimated losses were greater for the Medicare patients alone. The surgeon work effort was dramatically higher for the tertiary care procedures, whereas the reimbursement per work effort was lower. A 15% reduction in reimbursement would result in an estimated net loss to the hospital for each of the tertiary care procedures and would exacerbate the estimated losses to the department. CONCLUSIONS: Caring for complex surgical problems is currently profitable to an academic hospital but is associated with marginal losses for a department of surgery. Economic forces resulting from further decreases in hospital and physician reimbursement may limit access to academic medical centers and surgeons for patients with complex surgical problems and may compromise the overall academic mission. PMID- 10816630 TI - Cost-efficacy of cultured epidermal autografts in massive pediatric burns. AB - OBJECTIVE: To assess the efficacy of cultured epidermal autografts (CEA) for closure of burn wounds in pediatric burn patients with full-thickness burns of more than 90% total body surface area. SUMMARY BACKGROUND DATA: Paucity of donor sites in massive burns makes the use of expanded skin of paramount importance. CEA techniques have been used in burned patients with differing and controversial results. The true impact and the efficacy of such techniques in massive burns remain uncertain. METHODS: Patients with full-thickness burns of more than 90% body surface area treated between May 1988 and May 1998 were studied. Patients grafted with CEA were compared with patients grafted with conventional meshed autografts. Rates of death and complications, length of hospital stay (LOS), hospital cost, acute readmissions for reconstruction, and quality of scars were studied as outcome measures. RESULTS: Patients treated with CEA had a better quality of burn scars but incurred a longer LOS and higher hospital costs. Both groups had comparable readmissions for open wounds, but patients treated with CEA required more reconstructive procedures during the first 2 years after the injury. The incidence of sepsis and pneumonia in both groups was comparable. CONCLUSIONS: Conventional meshed autografts are superior to CEA for containing hospital cost, diminishing LOS, and decreasing the number of readmissions for reconstruction of contractures. However, the use of CEA provides better scar quality such that perhaps future research should focus on bioengineered dermal templates to promote take and diminish long-term fragility. PMID- 10816631 TI - Association of infiltrating lobular carcinoma with positive surgical margins after breast-conservation therapy. AB - OBJECTIVE: To determine whether infiltrating lobular carcinoma (ILC) is associated with high positive-margin rates for single-stage lumpectomy procedures, and to define clinical, mammographic, or histologic characteristics of ILC that might influence the positive-margin rate, thereby affecting treatment decisions. SUMMARY BACKGROUND DATA: Infiltrating lobular cancer represents approximately 10% of all invasive breast carcinomas and is often poorly defined on gross examination. METHODS: A group of 47 patients with biopsy-proven ILC undergoing breast-conservation therapy (BCT) at the University of Virginia Health Sciences Center between 1975 and 1999 was compared with a group of 150 patients with infiltrating ductal cancer undergoing BCT during the same time period. The pathology of the lumpectomy specimen was reviewed for each patient to confirm surgical margin status. Office and surgical notes as well as mammography reports were examined to determine whether the lesions were deemed palpable before and during surgery. Patients were stratified according to age, family history, tumor size, tumor location, and histologic features of the tumor. RESULTS: The incidence of positive margins was greater in the ILC group compared with the infiltrating ductal cancer group. Patient age, family history, and preoperative palpability of the tumor did not correlate with surgical margin status. Of the mammographic features identified, including spiculated mass, calcifications, architectural distortion, and other densities, only architectural distortion predicted positive surgical margin status. Tumor grade, tumor size, lymph node status, and receptor status were not predictive of surgical margin status. CONCLUSIONS: For patients with ILC, BCT is feasible, but these patients are at high risk of tumor-positive resection margins (51% incidence) after the initial resection. Only the mammographic finding of architectural distortion was identified as a preoperative marker reliably identifying a subgroup of ILC patients at especially high risk for a positive surgical margin. For all patients with ILC considering BCT, careful counseling about the potential need for a second procedure to treat the positive margin should be included in the treatment discussion. PMID- 10816632 TI - Patient compliance is critical for equivalent clinical outcomes for breast cancer treated by breast-conservation therapy. AB - OBJECTIVE: To determine the compliance with a standard breast-conservation therapy (BCT) program in a predominantly indigent, minority population of patients with early breast cancer (stage I and II) served by a rural state institution in the South; to compare the clinical outcomes of this group with those reported in clinical trials; and to examine the socioeconomic factors that may have contributed to the rate of compliance. SUMMARY BACKGROUND DATA: Disease free survival and overall survival in early breast cancer treated by BCT versus modified radical mastectomy are reported to be equivalent in prospective randomized trials. However, patients enrolled in clinical trials may not be representative of patients living in the various diverse communities that make up the United States. The authors' hypothesis is that patients enrolled in clinical trials at the national level may not be representative of indigent patients in the rural South and that clinical trial results may not be directly applicable. METHODS: A retrospective review of 55 women with early-stage breast cancer treated from 1990 to 1995 was performed. Clinical data, compliance with treatment and clinical follow-up, and recurrence rates were examined. Statistical analysis performed include the Fisher exact test, Kaplan-Meier survival analysis, and log rank test. RESULTS: Full compliance (defined as completion of the entire course of radiation therapy and clinical follow-up) with the BCT program was observed in only 36% of patients. Fifteen of the 35 noncompliant patients did not complete radiation therapy. A significantly higher local failure rate was observed: 8 of these 15 patients (53%) have had local failure. In contrast, patients who were either in full compliance with the BCT program or were deficient only in that they missed part of their clinical follow-up had local failure rates of 5% (1/20) and 10% (2/20), respectively. Age, race, stage of cancer, economic status (measured by availability of medical insurance), distance of patient's residence from the hospital, and education level were evaluated as potential predictors of compliance. None predicted patient compliance, although a trend toward higher compliance was noted in patients with a higher education level, as determined by literacy testing. CONCLUSIONS: Compliance with the BCT protocol at the authors' institution was worse than reported in clinical trials, and noncompliance translated into a significant increase in the local failure rate. Factors examined suggest that literacy may play a role in predicting compliance. Although BCT should be discussed with all breast cancer patients, the judicious application of clinical trial data to an institution's local population is warranted. PMID- 10816633 TI - Quality of life and outcomes after pancreaticoduodenectomy. AB - OBJECTIVE: To assess the quality of life (QOL) and functional outcome of patients after pancreaticoduodenectomy. SUMMARY BACKGROUND DATA: Pancreaticoduodenectomy is gaining acceptance and is being performed in increasing numbers for various malignant and benign diseases of the pancreas and periampullary region. There is a general impression that pancreaticoduodenectomy can severely impair QOL and alter normal activities. Only a few small studies have evaluated QOL after pancreaticoduodenectomy. METHODS: A standard QOL questionnaire was sent to 323 patients surviving pancreaticoduodenectomy who had undergone surgery at The Johns Hopkins Hospital between 1981 and 1997. Thirty items on a visual analog scale were categorized into three domains: physical (15 items), psychological (10 items), and social (5 items). Scores are reported as a percentile, with 100% being the highest possible score. The same QOL questionnaire was also sent to laparoscopic cholecystectomy patients and healthy controls. A separate component of the questionnaire asked about functional outcomes and disabilities. RESULTS: Overall QOL scores for the 192 responding pancreaticoduodenectomy patients in the three domains (physical, psychological, social) were 78%, 79%, and 81%, respectively. These QOL scores were comparable to those of the 37 laparoscopic cholecystectomy patients and the 31 healthy controls. The pancreaticoduodenectomy patients were subgrouped into chronic pancreatitis, other benign disease, pancreatic adenocarcinoma, and other cancers. Patients who underwent resection for chronic pancreatitis and pancreatic adenocarcinoma had significantly lower QOL scores in the physical and psychological domains compared with the laparoscopic cholecystectomy patients and the healthy controls. Common problems after pancreaticoduodenectomy were weight loss, abdominal pain, fatigue, foul stools, and diabetes. CONCLUSIONS: This is the largest single-institution experience assessing QOL after pancreaticoduodenectomy. These data demonstrate that as a group, patients who survive pancreaticoduodenectomy have near-normal QOL scores. Many patients report weight loss and symptoms consistent with pancreatic exocrine and endocrine insufficiency. Most patients have QOL scores comparable to those of control patients and can function independently in daily activities. PMID- 10816634 TI - Surgical management of recurrent pancreatitis in children with pancreas divisum. AB - OBJECTIVE: To analyze an institutional experience with pancreatitis in childhood to clarify the frequency of pancreas divisum in that patient population, the characteristics of pancreatitis in children with pancreas divisum, and the role of surgical management in their treatment. SUMMARY BACKGROUND DATA: The role of pancreas divisum in causing acute and relapsing pancreatitis and chronic, recurring abdominal pain is controversial. Although the anatomical abnormality is present from birth, most investigators have reported cases with onset of symptoms in adulthood. The reported pediatric experience with this disorder is small, and the natural history of pancreatitis in children with pancreas divisum has not been well elucidated. METHODS: A retrospective chart review of all children 18 years of age and younger with a discharge diagnosis of pancreatitis identified 135 patients treated in the authors' institution from 1978 to 1998. Ten patients were found to have anatomical variants of pancreas divisum associated with recurrent or chronic pancreatitis. The medical records of these patients were reviewed for data on the presentation, diagnostic findings, imaging studies, treatment, surgical findings, and pathologic findings in these children. Chart review and telephone calls were used to assess the current state of health in nine patients available for follow-up. RESULTS: Pancreas divisum was identified in 7.4% of all children with pancreatitis and 19.2% of children with relapsing or chronic pancreatitis. Patients had early onset of recurrent episodic epigastric pain and vomiting, at a mean age of 6 years. Three patients had a positive family history of pancreatitis and one was proven by DNA analysis to have hereditary pancreatitis. Pancreatitis was documented by elevated amylase or lipase levels, and endoscopic retrograde cholangiopancreatography was the method of diagnosis of pancreas divisum in all patients. Eight patients had complete pancreas divisum and two had incomplete variants. Eight patients underwent surgery to improve ductal drainage. Seven underwent transduodenal sphincteroplasty of the accessory papilla, along with sphincteroplasty of the major papilla in two (plus septoplasty in one). Three patients underwent longitudinal pancreaticojejunostomy, as a primary procedure in one patient with midductal stenosis and in two because of recurring pancreatitis after sphincteroplasty. The surgical findings and histologic examination of five patients undergoing distal pancreatectomy revealed striking changes of advanced chronic pancreatitis. Patients responding to sphincteroplasty alone showed less severe histologic changes. Overall, three of seven patients had excellent results, three were improved, and one had continued disabling attacks of pancreatitis. The mean duration of follow-up was 7.3 years, and there were no deaths. No patients had endocrine or exocrine pancreatic insufficiency, and none required chronic analgesics. CONCLUSIONS: Pancreas divisum is an important cause of recurrent pancreatitis in childhood and should be sought aggressively in children with more than one episode of pancreatitis or pancreatitis with a history of chronic recurrent abdominal pain. Surgical intervention is directed toward relief of ductal obstruction and may involve accessory duct sphincteroplasty alone or in combination with major sphincteroplasty and septoplasty. Patients with more distal ductal obstruction or ductal ectasia may benefit from pancreaticojejunostomy. PMID- 10816635 TI - Duodenopancreatic resections in patients with multiple endocrine neoplasia type 1. AB - OBJECTIVE: To review the authors' 7-year experience with a surgical approach for pancreatic and duodenal neuroendocrine tumors (NETs) in patients with multiple endocrine neoplasia type 1 (MEN 1) designed to remove all gross tumor with limited complications, preserving pancreatic function. SUMMARY BACKGROUND DATA: MEN 1 is an autosomal dominant familial neoplasia syndrome characterized by the development of NETs of the duodenum and pancreas. Some tumors are clinically insignificant or follow a benign course, although a subset pursues a malignant, lethal natural history; the risk of surgical management must be appropriate to the disease course. METHODS: The clinical, biochemical, genetic, and pathologic data were retrospectively reviewed for 21 consecutive MEN 1 patients undergoing pancreatic resection for NETs between 1993 and 1999 at one institution. Age at operation, presenting symptoms, results of preoperative and intraoperative localization studies, major and minor complications, and pathology, including metastases, were analyzed. RESULTS: The surgical approach was selected based on the location and size of the tumors. Five patients required pancreaticoduodenectomy, 11 patients underwent non-Whipple pancreatic resections, and 5 underwent simple enucleation of benign NETs. The incidence of regional lymph node metastases was 33%. CONCLUSIONS: Major pancreatic procedures can be performed safely in most patients with MEN 1 and NETs. Because NETs are the most common MEN 1-related cause of death in the authors' kindreds, an aggressive surgical approach, including early intervention before malignant spread and major pancreatic resection where indicated, appears justified. PMID- 10816636 TI - Functional outcomes after ileal pouch-anal anastomosis for chronic ulcerative colitis. AB - OBJECTIVE: To assess long-term outcomes after ileal pouch-anal anastomosis (IPAA) for chronic ulcerative colitis (CUC) with specific emphasis on patient sex, childbirth, and age. SUMMARY BACKGROUND DATA: Childbirth and the process of aging affect pelvic floor and anal sphincter function independently. Early function after IPAA is good for most patients. Nonetheless, there are concerns about the impact of the aging process as well as pregnancy on long-term functional outcomes after IPAA. METHODS: Functional outcomes using a standardized questionnaire were prospectively assessed for each patient on an annual basis. RESULTS: Of the 1,454 patients who underwent IPAA for CUC between 1981 and 1994, 1,386 were part of this study. Median age was 32 years. Median length of follow-up was 8 years. Pelvic sepsis was the primary cause of pouch failure irrespective of sex or age. Functional outcomes were comparable between men and women. Eighty-five women who became pregnant after IPAA had pouch function, which was comparable with women who did not have a child. Daytime and nocturnal incontinence affected older patients more frequently than younger ones. Incontinence became more common the longer the follow-up in older patients, but this was not found in younger patients. Poor anal function led to pouch excision in only 3 of 204 older patients. CONCLUSIONS: Incontinence rates were significantly higher in older patients after IPAA for CUC compared with younger patients. However, this did not contribute to a greater risk of pouch failure in these older patients. Patient sex and uncomplicated childbirth did not affect long-term functional outcomes. PMID- 10816637 TI - The proopiomelanocortin system in cutaneous neuroimmunomodulation. An introductory overview. PMID- 10816638 TI - The proopiomelanocortin system. AB - POMC (31,000 MW) is localized to the pituitary, brain, skin, and other peripheral sites. The particular enzyme profile present within a cell dictates the nature of the hormonal ligand (melanocortin) synthesized and secreted: melanotropic peptides (alpha-MSH beta-lipotropin, lambda-MSH), corticotropin (ACTH), several endorphins (e.g., met-enkephalin). These POMC-derived peptides mediate their actions through typical seven-spanning membrane receptors (MCRs; MCR1, 2, 3, 4, and 5). A specific melanocortin acting on a specific MCR regulates a particular biological response; for example, alpha-MSH on MCR1 increases melanogenesis within melanocytes, ACTH on MCR2 increases cortisol production within adrenal zona fasciculata cells. Within the brain melanocortins regulate satiety (MCR4) and erectile activity (MCR?). MCRs have been localized by melanocortin macromolecular probes, for example, fluorescent to human epidermal melanocytes and also to keratinocytes, suggesting that systemic melanocortins or localized POMC products might regulate these integumental cellular elements in synchrony to enhance skin pigmentation and/or immunological responses. Superpotent, prolonged acting melanotropic peptides have been synthesized and their application in clinical medicine has been demonstrated. MCR antagonists have been used to discover and further delineate other roles of melanocortin ligands. For example, melanocortin-induced satiety can be antagonized by a melanocortin antagonist. Defects in melanocortin ligand biosynthesis, secretion, and melanocortin receptor function can lead to a diverse number of pathological states. PMID- 10816639 TI - POMC-derived peptides and their biological action. AB - It has long been known that a large number of POMC-related peptides are found in skin. In this introduction I describe the formation of POMC-derived peptides in various tissues to indicate that processing is largely tissue-dependent. I focus on the peptides from the N-terminal fragment, such as gamma-MSH, ACTH and alpha MSH, and beta-lipopropin as well as beta-endorphin. I touch on the factors that control the synthesis of the various peptides, which are now numerous and varied, and again are tissue specific. The biologic activity of the peptides generated from POMC are described in relation to their possible action in skin. In addition, I describe a new class of peptides induced in skin following injury and which are of great interest. PMID- 10816640 TI - The pituitary-skin connection in amphibians. Reciprocal regulation of melanotrope cells and dermal melanocytes. AB - In amphibians, alpha-MSH secreted by the pars intermedia of the pituitary plays a pivotal role in the process of skin color adaptation. Reciprocally, the skin of amphibians contains a number of regulatory peptides, some of which have been found to regulate the activity of pituitary melanotrope cells. In particular, the skin of certain species of amphibians harbours considerable amounts of thyrotropin-releasing hormone, a highly potent stimulator of alpha-MSH release. Recently, we have isolated and sequenced from the skin of the frog Phyllomedusa bicolor--a novel peptide named skin peptide tyrosine tyrosine (SPYY), which exhibits 94% similarity with PYY from the frog Rana ridibunda. For concentrations ranging from 5 x 10(-10) to 10(-7) M, SPYY induces a dose-related inhibition of alpha-MSH secretion. At a dose of 10(-7) M, SPYY totally abolished alpha-MSH release. These data strongly suggest the existence of a regulatory loop between the pars intermedia of the pituitary and the skin in amphibians. PMID- 10816641 TI - The subtilisin/kexin family of precursor convertases. Emphasis on PC1, PC2/7B2, POMC and the novel enzyme SKI-1. AB - Proopiomelanocortin (POMC) is a precursor to various, bioactive peptides including ACTH, beta LPH, alpha MSH, and beta endorphin (beta END). Processing of POMC at dibasic residues is tissue-specific and is performed by either PC1 alone (resulting in ACTH and beta LPH, anterior pituitary corticotrophes) or by a combination of PC1 and PC2 (yielding alpha MSH and beta END, pituitary neurointermediate lobe and hypothalamus). The PC2-specific binding protein 7B2 is intimately involved in the zymogen activation of proPC2 into PC2. Structure function studies of these enzymes demonstrated the presence of N- and C-terminal domains, as well as specific amino acids within the catalytic segment that influence the degree of activity of each enzyme and the interaction of PC2 with 7B2. The tissue distribution, plasticity of expression, and the multiple precursors that are differentially cleaved by PC1 and/or PC2, predict a wide array of combinatorial activities of these convertases within the endocrine and neuroendocrine system. The phenotypic consequences of the absence of genetic expression of either PC1 or PC2 are now explored using knockout mice and in human patients suffering from obesity and diabetes. PMID- 10816642 TI - The role of melanocortins in adipocyte function. AB - It has been demonstrated that adipocytes express high affinity ACTH and alpha-MSH binding sites, and that ACTH, alpha-MSH, and beta-LPH are potent lipolytic hormones. Considerable species variability exists in the lipolytic response to melanocortins, however. Recently, MC2 and MC5 receptor-mRNA was found in both murine adipocytes and in the 3T3-L1 murine embryonic fibroblast cell line, but only after the 3T3-L1 cells had differentiated into adipocytes. The 3T3-L1 cell line was used to characterize the pharmacological properties of both MC2 and MC5 receptors in situ. Both murine MC2 and MC5 receptors are functional in the adipocyte, although the MC5 receptor required high doses of alpha-MSH to activate cylase. ACTH potently stimulates cyclase with EC50 values that are consistent with the hypothesis that the murine MC2 receptor, not the MC5 receptor, mediates stress-induced lipolysis via release of ACTH from the pituitary. PMID- 10816643 TI - An immunocytochemical approach to the study of beta-endorphin production in human keratinocytes using confocal microscopy. AB - Proopiomelanocortin (POMC) is a protein that is posttranslationally processed to yield POMC peptides. The main site of POMC expression is the anterior pituitary lobe but many other sources have been identified. There is evidence that the skin produces POMC peptides, although their roles have not yet been defined. In the skin, regulation of POMC gene expression is known to be hair-cycle dependent, and it is localized to the sebaceous gland. In particular, beta-endorphin, a POMC peptide, has been shown to be modulated by TPA, IL-1 alpha, and ultraviolet radiation in keratinocytes. These results were obtained by examination of POMC mRNA levels using the Northern blot method; beta-endorphin protein production by the Western blot method on cultured cells; and immunocytochemistry for tissue preparations. This report represents an approach to use immunocytochemistry to quantify beta-endorphin production in cultured human keratinocytes. Additionally, we examined whether exposure to 20 mJ ultraviolet B radiation (UVB) and/or UVA could influence beta-endorphin production in these cells. Keratinocytes were grown in monolayers, in serum-free medium, fixed, and incubated with antiserum to whole synthetic beta-endorphin. Fluorescence microscopy was performed with a confocal laser scanning microscope. The integrated level of fluorescence was evaluated in n = 18 +/- 8 individual cells, and this was assumed to be proportional to beta-endorphin content. High variability was observed in the fluorescence intensity among cells. No significant differences between control and UVB- or UVA + UVB-treated cells was found. Similar results were produced by using brefeldin A, a compound that disrupts the secretory pathway, eliminating the possibility that the absence of a difference between beta-endorphin content in the treated and control cells was due to secretion of the peptide into the medium. We conclude that: (1) beta-endorphin or beta-endorphin-like peptides are produced in human keratinocytes and are readily detected by immunocytochemistry; (2) under the conditions tested, UVA and/or UVB did not increase beta-endorphin like immunoreactivity in these cells. PMID- 10816644 TI - UV light and MSH receptors. AB - Ultraviolet B (UVB) radiation in the skin induces pigmentation that protects cells from further UVB damage and reduces photocarcinogenesis. Although the mechanisms are not well understood, our laboratory has shown that UVB radiation causes increased MSH receptor activity by redistributing MSH receptors from internal pools to the external surface, with a resultant increase in cellular responsiveness to MSH. By this means, UVB and MSH act synergistically to increase melanin content in the skin of mice and guinea pigs. In humans, MSH causes increased skin pigmentation, predominantly in sun-exposed areas. We have shown recently that UVB irradiation and exposure to MSH or to dbcAMP, stimulates production of mRNAs for both alpha MSH receptors and POMC in human melanocytes and keratinocytes. This indicates that at least one action of UVB on the pigmentary system is mediated through increased MSH receptor production, as well as through the production of the signal peptides, MSH and ACTH, that can further activate MSH receptors. The results add support to the hypothesis that the effects of UVB on cutaneous melanogenesis are mediated through a series of coordinated events in which MSH receptors and POMC-derived peptides play a central role. PMID- 10816645 TI - The melanocortin-1 receptor and human pigmentation. AB - alpha-Melanocyte stimulating hormone (alpha-MSH) is known to be the main physiologic regulator for integumental pigmentation of various vertebrate species. However, the role of alpha-MSH and related melanocortins in the regulation of human cutaneous pigmentation is only beginning to be understood. Cloning of the melanocortin-1 receptor (MC1R), and the feasibility of establishing normal human epidermal melanocyte cultures have made it possible to demonstrate direct and specific biological effects of alpha-MSH on these cells. It is now recognized that both alpha-MSH and ACTH have similar mitogenic and melanogenic effects on human epidermal melanocytes. These effects are mediated by binding of these hormones to the specific MC1R that recognizes them both with similar affinity. Human MC1R is homologous to its mouse counterpart in that its activation leads to stimulation of eumelanin synthesis. MC1R is also the binding site for agouti signaling protein (ASP), the product of the agouti locus. Human epidermal melanocytes respond to purified recombinant mouse or human ASP, with a reduction in basal tyrosinase activity, and complete abrogation of the mitogenic and melanogenic effects of alpha-MSH. These results suggest that ASP induces pheomelanin synthesis by competing with alpha-MSH for binding to the MC1R. This receptor seems to be subject to regulation by a variety of paracrine and/or autocrine factors that are synthesized in response to exposure of the skin to ultraviolet radiation (UVR). Activation of MC1R seems to be pivotal for UV induced melanogenesis, since stimulation of the cAMP pathway plays a key role in the melanogenic response of human epidermal melanocytes. The melanogenic response to UVR might be influenced by the presence of allelic variants of the MC1R gene. Allelic variants have been identified and shown to be associated with red hair, poor tanning ability, and possibly melanoma. The possible influence of these variants on the function of the MC1R needs to be investigated, in order to understand the physiological consequence of these mutations. Also, the interaction of alpha-MSH with other factors that are known to affect pigmentation needs to be better understood in order to define the role possible of this hormone and its receptor in acquired human cutaneous hyper- or hypopigmentation. PMID- 10816646 TI - Genetic studies of the human melanocortin-1 receptor. AB - Genetic approaches have suggested a critical role for the melanocortin-1 receptor in the control of pigmentation. We showed that this gene is unusually polymorphic in European populations and that, of the many variants, three in particular appear to be associated with red hair or fair skin. Family studies suggest these are inherited as an autosomal recessive trait (or at least approximate to this in many families). To date all individuals with two of these three changes (homozygote or compound heterozygote) have red hair. Early functional studies are in keeping with defective signalling through MC1R. An interested and perhaps unexpected question relates to the evolutionary factors that have given rise to such variants. Two models can be proposed, that are based on multiple alleles with minor changes in function or genetic hitch-hicking. PMID- 10816647 TI - Molecular pharmacology of Agouti protein in vitro and in vivo. AB - Agouti protein and Agouti-related protein (Agrp) are paracrine signaling molecules that act by antagonizing the effects of melanocortins, and several alternatives have been proposed to explain their mechanisms of action. Genetic crosses in a sensitized background uncover a phenotypic difference between overexpression of Agouti and loss of Mc1r function, demonstrate that a functional Mc1r is required for the pigmentary effects of Agouti, and suggest that Agouti protein can act as an agonist of the Mc1r in a way that differs from alpha-MSH stimulation. In vitro, Agouti protein inhibits melanocortin action by two mechanisms: competitive antagonism that depends on the carboxyterminus of the protein, and downregulation of melanocortin receptor signaling that depends on the aminoterminus. Our findings provide evidence of a novel signaling mechanism whereby alpha-MSH and Agouti protein function as independent ligands that inhibit each other's binding and transduce opposite signals through a single receptor. PMID- 10816648 TI - Synthetic melanocortin receptor agonists and antagonists. AB - Multiple melanocortin receptor subtypes with distinct cell and tissue distribution patterns have recently been identified, thereby presenting numerous opportunities for biological investigation. Many of these studies could benefit from the availability of subtype selective or specific agonists and antagonists. The purpose of this report is to summarize the state of available melanocortin receptor agonists and antagonists. PMID- 10816649 TI - Proopiomelanocortin and the immune-neuroendocrine connection. AB - This presentation will cover the history, recent developments in, and implications of the ability of both the immune and neuroendocrine systems to produce POMC. The discovery of POMC in immune cells was one of the events that heralded a molecular understanding of neuroimmunomodulation. This, together with the presence of opiate and ACTH receptors on lymphocytes and macrophages, provided the first biochemical circuit for which the same signal molecules and receptors could be used for intrasystem regulation, as well as bidirectional communication between the immune and neuroendocrine systems. Today we have a quite good understanding of the regulation and processing of POMC in immune cells, as well as the interaction of its product peptides with other cytokines. For instance, IL-1 causes POMC production by immune cells, and the POMC product, alpha-MSH, in turn, acts functionally as an IL-1 antagonist. In the past year, the expression of full-length POMC mRNA has been reported and this solved one of the paradoxes with respect to POMC production, processing, and secretion. We provide data on these developments together with quite startling findings on the physiologic function of POMC peptides in the immune system. Among these are the local antinociceptive effects of immune cell-derived beta-endorphin, altered hematopoiesis in opiate receptor-deficient animals, and the diagnosis of ACTH insensitivity by a deficiency of ACTH receptors on lymphocytes. PMID- 10816650 TI - Mechanisms of antiinflammatory action of alpha-MSH peptides. In vivo and in vitro evidence. AB - alpha-Melanocyte stimulating hormone (alpha-MSH) modulates all forms of inflammation by acting on peripheral inflammatory cells, glial inflammatory cells, and on CNS receptors that activate descending antiinflammatory neural pathways. The multiple actions of this ancient peptide suggest that there is no singular biochemical mechanism through which it exerts its antiinflammatory activity. However, research on IL-10 deficient and Agouti protein hypersecreting mice provide new insights into the actions of the peptide in living animals. Studies of cultured human astrocytes, whole murine brain, and human monocyte/macrophages indicate that a primary effect of the peptide is modulation of activation of the nuclear transcription factor kappa B. The latter influence may underlie the established reduction of gene expression and production of proinflammatory peptides and inducible nitric oxide by alpha-MSH peptides. PMID- 10816651 TI - alpha-MSH in systemic inflammation. Central and peripheral actions. AB - Until recently, inflammation was believed to arise from events taking place exclusively in the periphery. However, it is now clear that central neurogenic influences can either enhance or modulate peripheral inflammation. Therefore, it should be possible to improve treatment of inflammation by use of antiinflammatory agents that reduce peripheral host responses and inhibit proinflammatory signals in the central nervous system (CNS). One such strategy could be based on alpha-melanocyte stimulating hormone (alpha-MSH). Increases in circulating TNF-alpha and nitric oxide (NO), induced by intraperitoneal administration of endotoxin in mice, were modulated by central injection of a small concentration of alpha-MSH. Inducible nitric oxide synthase (iNOS) activity and iNOS mRNA in lungs and liver were likewise modulated by central alpha-MSH. Increase in lung myeloperoxidase (MPO) activity was significantly less in lungs of mice treated with central alpha-MSH. Proinflammatory agents induced by endotoxin were significantly greater after blockade of central alpha-MSH. The results suggest that antiinflammatory influences of neural origin that are triggered by alpha-MSH could be used to treat systemic inflammation. In addition to its central influences, alpha-MSH has inhibitory effects on peripheral host cells, in which it reduces release of proinflammatory mediators. alpha-MSH reduces chemotaxis of human neutrophils and production of TNF-alpha, neopterin, and NO by monocytes. In research on septic patients, alpha-MSH inhibited release of TNF-alpha, interleukin-1 beta (IL-1 beta), and interleukin-8 (IL-8) in whole blood samples in vitro. Combined central and peripheral influences can be beneficial in treatment of sepsis. PMID- 10816652 TI - Human peripheral blood-derived dendritic cells express functional melanocortin receptor MC-1R. AB - The neuropeptide, alpha-melanocyte-stimulating hormone (alpha-MSH) is well known for its immunomodulating capabilities. alpha-MSH antagonizes the activity of numerous proinflammatory mediators; for example, Interleukin-1 (IL-1), IL-6, tumor necrosis factor alpha (TNF alpha), and bacterial endotoxin. In vivo alpha MSH has been shown to suppress a contact hypersensitivity reaction in mice, and to induce hapten-specific tolerance. Since antigen presenting cells (APC) represent key elements for tolerance induction, the effect of alpha-MSH, and the expression of its receptor-melanocortin receptor-1 (MC-1R), on human peripheral blood-derived monocytes and dendritic cells (DC), was investigated. Semiquantitative RT-PCR demonstrated that monocytes and DC express MC-1R, but none of the other members of the MC-receptor family. Moreover, the extent of MC 1R expression correlated with the state of activation of these cells. Since the major ligand of MC-1R is alpha-MSH the question of whether alpha-MSH affects the function of monocyte derived DC was further investigated. We found that the expression of the costimulatory molecules CD 86 and CD 40 was downregulated on DC in the presence of alpha-MSH. Thus, alpha-MSH may exert its immunosuppressive effects by altering the function of APC. PMID- 10816653 TI - Neural influences on induction of contact hypersensitivity. AB - Contact hypersensitivity (CH)-induction begins when cutaneous antigen-presenting cells (APC) capture hapten that has been applied epicutaneously, and the process prepares hapten for presentation to T-cells. APCs are functionally plastic, are influenced by the microenvironment in which they reside, and their functional properties have a profound effect on the phenotype of the hapten-specific T-cells that they activate. Ultraviolet B radiation (UVR) distorts the cutaneous microenvironment, thereby altering local APC function, and changing the immune outcome from sensitization to unresponsiveness. Although UVR induces keratinocytes to produce TNF alpha and IL-10 (cytokines that have been implicated in failed CH-induction and tolerance, respectively, after UVR), dermal mast cells turn out to be the source of these immunomodulatory cytokines. Mast cell degranulation is triggered by CGRP released from UVR-exposed cutaneous nerve termini. Even in normal skin, cutaneous nerves influence the immune response to haptens. Substance P released from cutaneous nerves acts as an adjuvant, raising the immunogenicity of epicutaneously applied haptens. Thus, the nerves and the neuropeptides that these processes release contribute to the cutaneous microenvironment. By altering APC function, cutaneous nerves can dictate the quality and the quantity of immune responses to antigens of the skin. PMID- 10816654 TI - Role of epidermal cell-derived alpha-melanocyte stimulating hormone in ultraviolet light mediated local immunosuppression. AB - Irradiation of the skin with ultraviolet light (UV) results in profound alterations of both local and systemic immune responses. These effects are largely mediated by soluble mediators released from epidermal cells in response to UV. It is well known that keratinocytes release increased amounts of cytokines upon UV-irradiation. UV-light also induces the release of the proopiomelanocortin (POMC)-derived peptide, alpha-melanocyte-stimulating hormone (alpha MSH), from keratinocytes, and upregulates the expression of POMC mRNA. alpha MSH exerts a variety of immunomodulating and antiinflammatory effects, mainly by virtue of its capacity to alter the function of antigen presenting cells and vascular endothelial cells. Within an in vivo mouse-model, both intravenous and topical application of alpha MSH resulted in inhibiting the induction, eliciting a contact hypersensitivity reaction, and inducing hapten-specific tolerance. These findings indicate that alpha MSH, released in the epidermis after UV irradiation, may contribute to UV-mediated immunosuppression. The therapeutic application of alpha MSH or alpha MSH-derived peptides may prove to be a useful approach for treating inflammatory skin diseases. PMID- 10816655 TI - alpha-MSH and the regulation of melanocyte function. AB - alpha-MSH, has numerous actions in the skin and by activating the MC1 receptor (MC1-R) on melanocytes it stimulates melanogenesis. Rather than producing large increase in melanin production alpha-MSH acts specifically to stimulate eumelanin synthesis. Although this could be important in determining skin color and tanning there is debate as to the pigmentary significance of alpha-MSH in humans. Circulating levels of alpha-MSH are negligible and although it is produced in the skin by different cell types, including melanocytes, the major skin form is desacetyl alpha-MSH, and this is a weak agonist at MC1-R. Certain ACTH peptides, notably ACTH1-17, are more potent agonists at the MC1-R and, since their skin concentrations exceed those of alpha-MSH, they could serve as natural ligands at this receptor and regulate pigmentary responses in humans. Activation of MC1-R does, however, produce other responses in human melanocytes. Thus, alpha-MSH stimulates melanocyte dendricity and attachment to extracellular matrix proteins. It also protects melanocytes from the damaging effects of oxidative stress, and regulates their production of NO by modulating the induction of iNOS--as it does within macrophages. alpha-MSH clearly affects various aspects of melanocyte behavior and its melanogenic effects could be the consequence of a more fundamental role in the melanocyte. The precise nature of this role is unclear, but it could be part of a generic role that alpha-MSH and other POMC peptides have in skin homeostasis. PMID- 10816656 TI - Molecular basis of the alpha-MSH/IL-1 antagonism. AB - The neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) is recognized as a potent mediator of immune and inflammatory reactions. Accordingly, alpha-MSH in vitro, as well as in vivo, antagonizes the proinflammatory activities of cytokines such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor alpha (TNF alpha). Since the molecular basis of these antiinflammatory effects is not well known, the influence of alpha-MSH on IL-1 beta-induced chemokine production and transcription factor activation was investigated in human keratinocytes. alpha-MSH, in a dose-dependent manner, after 48 h, significantly reduced the IL-1 beta mediated secretion of the C-X-C chemokines IL-8 and Gro alpha. This was confirmed by semiquantitative RT-PCR, which revealed a marked down-regulation in IL-8 and Gro alpha mRNA expression. Furthermore, we determined the effect of alpha-MSH on the IL-1 beta-induced activation of the nuclear factor kappa B (NF kappa B)--a major transcription factor for chemokine genes. Electrophoretic mobility-shift-assays showed that alpha-MSH, in a dose range from 10(-6) to 10( 12) M, significantly downregulated the IL-1 beta-induced activation of NF kappa B 10 minutes after stimulation. Therefore, NF kappa B inactivation by alpha-MSH appears to be a crucial event, one that is responsible for the downregulation of cytokine gene transcription. PMID- 10816657 TI - Expression of functional melanocortin receptors and proopiomelanocortin peptides by human dermal microvascular endothelial cells. AB - Human dermal microvascular endothelial cells (HDMEC) are capable of mediating leukocyte-endothelial interactions by the expression of cellular adhesion molecules and the release of proinflammatory cytokines and chemokines during cutaneous inflammation. Recent studies support the important role for proopiomelanocortin (POMC) peptides, such as alpha-melanocyte stimulating hormone (alpha-MSH), as immunomodulators in the cutaneous immune system. The purpose of the studies described here was to determine whether HDMEC serves as both target and source for POMC peptides. RT-PCR and Northern blot studies demonstrated the constitutive expression of mRNA for the adrenocorticotropin (ACTH) and alpha-MSH specific melanocortin receptor 1 (MC-1R) in HDMEC, and the microvascular endothelial cell line HMEC-1 that could be upregulated by stimulation with IL-1 beta and alpha-MSH. HDMEC responded to stimulation by alpha-MSH with a dose- and time-dependent synthesis and release of the CXC chemokines, IL-8 and GRO alpha. Likewise, alpha-MSH augmented HDMEC chemokine release induced by TNF or IL-1. HD MEC were found to constitutively express POMC and prohormone convertase 1 (PC-1); the latter being required to generate ACTH from the POMC prohormone. POMC and PC 1 mRNA expression are increased as a result of stimulation with UVB and UVA1 radiation, IL-1, and alpha-MSH. In addition, UV-radiation is capable of inducing the release of HDMEC, ACTH, and alpha-MSH in a time- and dose-dependent fashion. Thus, these data provide evidence that HDMEC are capable of expressing functional MC-1R, POMC, and PC-1 mRNA; and of releasing POMC peptides with UV light, IL-1, and alpha-MSH as regulatory factors. The expression and regulation of these peptides may be of importance, not only for the autocrine or paracrine regulation of physiologic functions of dermal endothelial cells, but also for the regulation of certain microvascular-mediated cutaneous or systemic inflammatory responses. PMID- 10816658 TI - Mechanisms of the antiinflammatory effects of alpha-MSH. Role of transcription factor NF-kappa B and adhesion molecule expression. AB - The recruitment of leukocytes from the circulation to inflamed tissue is regulated by the expression of adhesion molecules on both leukocytes and endothelial cells. The proopiomelanocortin-derived peptide alpha-melanocyte stimulating hormone (alpha-MSH) is known to modulate inflammation. Thus, we investigated the influence of alpha-MSH on the LPS-induced expression of the adhesion molecules E-selectin and VCAM-1 on endothelial cells. Human microvascular endothelial cells (HMEC-1) were treated with LPS (100 ng/ml) alone or in the presence of alpha-MSH (10(-8) to 10(-16) M). RT-PCR analysis showed that alpha-MSH significantly reduced LPS-induced expression of VCAM-1 and E selectin. Since many adhesion molecules contain regulatory NF-kappa B sites in their promoter region, the role of alpha-MSH in the activation of the transcription factor NF-alpha B was also investigated. alpha-MSH significantly downregulated the LPS-mediated activation of NF-kappa B, in a dose-dependent manner. These findings indicate that modulation of the transcription factor NF kappa B is a crucial molecular event, one that seems to be responsible for the antiinflammatory effects of alpha-MSH. PMID- 10816659 TI - POMC and fibroblast biology. AB - We evidenced in vitro proopiomelanocortin (POMC) mRNA-transcription in human dermal fibroblasts using Northern blot hybridization. Modulation of POMC gene expression by cytokines (transforming growth factor-beta, TGF-beta, and tumor necrosis factor-alpha, TNF-alpha) was investigated by incubating human normal fibroblasts with 1 and 10 ng/ml cytokines, either alone or in combination, for 24 hours. Our results show that dermal fibroblasts express POMC at significant levels under unstimulated conditions. POMC steady-state levels were significantly reduced by addition of TGF-beta. On the other hand, TNF-alpha exerted a stimulatory effect on POMC mRNA transcription, partially counteracting the effect of TGF-beta. These data provide the first demonstration of POMC gene expression in cultured skin fibroblasts. The opposite regulatory effect of TGF-beta and TNF alpha, two cytokines primarily involved in extracellular matrix regulation, suggests a possible role for POMC-derived peptides in fibroblast activity. We also investigated POMC mRNA expression in keloid-derived fibroblasts in culture, and its regulation by TGF-beta added at the highest concentration documented for inhibition. Keloid-derived fibroblasts showed clearly detectable levels of POMC mRNA in basal conditions, and no alteration of POMC gene expression was observed when TGF-beta was added in culture. The altered TGF-beta regulation of POMC mRNA levels suggest that POMC-derived peptides may play a role in the pathogenesis of keloid formation through an autocrine/paracrine network, resulting in modulation of extracellular matrix synthesis. PMID- 10816660 TI - The role of proopiomelanocortin-derived peptides in skin fibroblast and mast cell functions. AB - We have previously described proopiomelanocortin (POMC) gene-expression in human normal cultured dermal fibroblasts, and its dose- and time-dependent modulation by transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha). The aim of the work described here was to investigate POMC-derived peptide release in vitro by cultured fibroblasts following incubation with different concentrations of both TNF-alpha and TGF-beta for 24 hours (1, 5, and 10 ng/ml). The effect of simultaneous addition of both TNF-alpha and TGF-beta (10 ng/ml) was also evaluated. Culture supernatants of human skin fibroblasts were collected to detect adrenocorticotropin hormone (ACTH), alpha-melanotropin (alpha MSH), and beta-endorphin (beta-EP) levels by specific immunoenzymatic assay. We investigated the in vitro histamine-releasing activity of the POMC-derived peptides, alpha-MSH and beta-EP, on human foreskin mast cells. Detection of cleavage products in supernatants from cultured normal human dermal fibroblasts indicated intracellular processing by POMC protein. We were able to measure detectable levels of all peptides in basal conditions. TNF-alpha addition resulted in an increase in beta-EP and ACTH levels. TGF-beta-stimulated fibroblasts showed no alteration in beta-EP and alpha-MSH levels, whereas ACTH release was significantly enhanced. Both alpha-MSH and beta-EP induced histamine release from human foreskin mast cells in vitro with beta-EP-induced histamine levels as high as those observed with the calcium ionophore, ionomycin. Our data document fibroblast POMC-derived peptide release and modulation by cytokines, suggesting that they have a possible role in extracellular matrix deposit regulation and skin inflammation. PMID- 10816661 TI - Alpha-melanocyte-stimulating hormone modulates activation of NF-kappa B and AP-1 and secretion of interleukin-8 in human dermal fibroblasts. AB - Alpha-melanocyte-stimulating hormone (alpha-MSH) has evolved as a mediator of diverse biological activities in an ever-growing number of non-melanocytic cell types. One mechanism by which alpha-MSH exerts its effects is modulation of AP-1 and NF-kappa B. These two transcription factors also play an important role in fibroblasts, in extracellular matrix composition, and in cytokine expression. By use of electric mobility shift assays, we demonstrate that alpha-MSH (10(-6) to 10(-14) M) activates AP-1 in human dermal fibroblasts, whereas coincubation with interleukin-1 beta (IL-1 beta) results in suppression of its activation. alpha MSH also induces activation of NF-kappa B but does not modulate DNA binding on costimulation with IL-1 beta. Since AP-1 and NF-kappa B are key elements in controlling interleukin-8 (IL-8) transcription, human fibroblasts were treated with alpha-MSH and IL-1 beta for 24 hours, and cytokine levels in the supernatants were measured by ELISA. alpha-MSH alone had little effect, whereas coincubation with IL-1 beta led to marked downregulation of IL-8 secretion (at most 288 +/- 152 ng/mL) when compared to treatment with IL-1 beta alone (919 +/- 157 ng/mL). Our results indicate that alpha-MSH exerts modulatory effects on the activation of NF-kappa B and AP-1, and that it can regulate chemokine secretion in human dermal fibroblasts. These effects of alpha-MSH may have important regulatory functions in extracellular matrix composition, wound healing, or angiogenesis. PMID- 10816662 TI - Cutaneous expression of CRH and CRH-R. Is there a "skin stress response system?". AB - The classical neuroendocrine pathway for response to systemic stress is by hypothalamic release of corticotropin releasing hormone (CRH), subsequent activation of pituitary CRH receptors (CRH-R), and production and release of proopiomelanocortin (POMC) derived peptides. It has been proposed that an equivalent to the hypothalamic-pituitary-adrenal axis functions in mammalian skin, in response to local stress (see Reference 1). To further define such system we used immunocytochemistry, RP-HPLC separation, and RIA techniques, in rodent and human skin, and in cultured normal and malignant melanocytes and keratinocytes. Production of mRNA for CRH-R1 was documented in mouse and human skin using RT-PCR and Northern blot techniques; CRH binding sites and CRH-R1 protein were also identified. Addition of CRH to immortalized human keratinocytes, and to rodent and human melanoma cells induced rapid, specific, and dose-dependent increases in intracellular Ca2+. The latter were inhibited by the CRH antagonist alpha-helical-CRH(9-41) and by the depletion of extracellular calcium with EGTA. CRH production was enhanced by ultraviolet light radiation and forskolin (a stimulator for intracellular cAMP production), and inhibited by dexamethasone. Thus, evidence that skin cells, both produce CRH and express functional CRH-R1, supports the existence of a local CRH/CRH-R neuroendocrine pathway that may be activated within the context of a skin stress response system. PMID- 10816663 TI - Corticotropin releasing factor receptors and their ligand family. AB - The CRF receptors belong to the VIP/GRF/PTH family of G-protein coupled receptors whose actions are mediated through activation of adenylate cyclase. Two CRF receptors, encoded by distinct genes, CRF-R1 and CRF-R2, and that can exist in two alternatively spliced forms, have been cloned. The type-1 receptor is expressed in many areas of the rodent brain, as well as in the pituitary, gonads, and skin. In the rodent, one splice variant of the type-2 receptor, CRF-R2 alpha, is expressed mainly in the brain, whereas the other variant, CRF-R2 beta, is found not only in the CNS, but also in cardiac and skeletal muscle, epididymis, and the gastrointestinal tract. The poor correlation between the sites of expression of CRF-R2 and CRF, as well as the relatively low affinity of CRF for CRF-R2, suggested the presence of another ligand, whose existence was confirmed in our cloning of urocortin. This CRF-like peptide is found not only in brain, but also in peripheral sites, such as lymphocytes. The broad tissue distribution of CRF receptors and their ligands underscores the important role of this system in maintenance of homeostasis. Functional studies of the two receptor types reveal differences in the specificity for CRF and related ligands. On the basis of its greater affinity for urocortin, in comparison with CRF, as well as its brain distribution, CRF-R2 may be the cognate receptor for urocortin. Mutagenesis studies of CRF receptors directed toward understanding the basis for their specificity, provide insight into the structural determinants for hormone receptor recognition and signal transduction. PMID- 10816664 TI - alpha-MSH can control the essential cofactor 6-tetrahydrobiopterin in melanogenesis. AB - In the human epidermis both keratinocytes and melanocytes express POMC m-RNA. Immunohistochemical studies of both cell types demonstrate significantly higher levels of alpha-MSH in melanocytes than in keratinocytes. Both cell types also hold the full capacity for de novo synthesis/recycling of the essential cofactor (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin (6BH4). 6BH4 is critical for the hydroxylation of the aromatic amino acids L-phenylalanine, L-tyrosine, and L tryptophan, for nitric oxide production and in various immune modulatory processes. Recently it was shown that tyrosinase activity is regulated by 6BH4 through a specific allosteric inhibition. The tyrosinase/6BH4 inhibition can be activated by 1:1 complex formation between 6BH4 and alpha-MSH, but an excess of alpha-MSH over 6BH4 can inhibit tyrosinase due to complex formation by tyr2 in the alpha-MSH sequence. In both melanocytes and keratinocytes 6BH4 controls the L tyrosine supply via phenylalanine hydroxylase (PAH). Recently we were able to show that the cellular uptake of L-phenylalanine and its intracellular turnover to L-tyrosine is crucial for melanogenesis. alpha-MSH can promote the production of L-tyrosine via PAH due to activation of the PAH tetramer to the more active dimer by removing 6BH4 from the regulatory binding domain on the enzyme. In conclusion, alpha-MSH can control (1) intracellular L-tyrosine formation from L phenylalanine in both melanocytes and keratinocytes, and (2) tyrosinase activity, directly, in melanocytes. PMID- 10816665 TI - Melanocortins and the treatment of nervous system disease. Potential relevance to the skin? AB - For several decades melanocortins have been implicated in the modulation of brain function. More recently, this idea has been supported by the identification and cloning of melanocortin (MC) receptors in the nervous system. MCs stimulate axonal growth in fetal neural tissue or in neural cell lines in culture. This feature was utilized in screening their neurotrophic or neuroprotection potential in animal studies of nervous system disease (peripheral nerve and spinal cord trauma, toxic and metabolic neuropathies, EAN, EAE, etc.). Some of these effects may be mediated by MC4 receptor activation, although as yet unknown receptors may also be involved (for instance, protection by Org 2766). To what extent MC nervous system effects are related to known effects of MCs in skin- and neuro immune systems, remains to be discovered. Nevertheless, it is of interest to note that activation of brain MC4 receptors profoundly affects care behavior for the body surface (skin and fur). The excessive grooming response in rodents exhibits a remarkable functional correlation with MSH activity in a brain-skin axis. PMID- 10816667 TI - Expression of MC1- and MC5-receptors on the human mast cell line HMC-1. PMID- 10816666 TI - The skin POMC system (SPS). Leads and lessons from the hair follicle. AB - Human and murine skin are prominent extrapituitary sources and targets for POMC products. The expression of, for example, ACTH, alpha-MSH, beta-endorphin, and MC 1-receptors fluctuates during synchronized hair follicle cycling in C57BL/6 mice. Since hair growth can be induced by ACTH injections in mice and mink, and since high doses of MSH peptides modulate epidermal and/or follicle keratinocyte proliferation in murine skin organ culture, some POMC products may operate as locally generated growth modulators, in addition to their roles in cutaneous pigment and immunobiology. Intrafollicularly generated ACTH and alpha-MSH as well as their cognate receptors may assist in the maintenance of the peculiar immune privilege of the anagen hair bulb. Possibly, they are also involved in the development of the follicle pigmentary unit, with whose generation their expression coincides. Given that murine skin also expresses (in a hair-cycle dependent way) CRH and CRH-R, which control pituitary POMC expression and in view of the fact that CRH arrests follicles in telogen, this suggests the existence of a local skin POMC system (SPS). This may be an integral component of cutaneous stress response-systems, and may most instructively be studied using the murine hair cycle as a model. PMID- 10816668 TI - Characterization of mu-opiate receptor in human epidermis and keratinocytes. PMID- 10816669 TI - Characterization of a polyclonal antibody raised against the human melanocortin-1 receptor. AB - We have generated a polyclonal antibody raised against a synthetic peptide corresponding to the amino acids 2-18 of the extracellular, N-terminal domain of the human melanocortin-1 receptor (MC-1R). Specificity of the affinity-purified anti-MC-1R antibody was confirmed by dot blot analysis with the antigenic peptide. The antibody detected MC-1R antigenicity on the surface of normal human melanocytes and WM35 melanoma cells, as shown by FACS and immunofluorescence analysis. The antibody was suitable for immunoperoxidase staining of deparaffinized skin sections, revealing prominent MC-1R staining of a cutaneous melanoma as opposed to undiseased skin in which normal melanocytes were only occasionally immunoreactive. Distinct adnexal structures in normal skin also displayed MC-1R immunostaining. Specificity of the MC-1R immunoreactivity in each technique was confirmed by preabsorption with the immunogenic peptide, omission, or substitution of the primary antibody with preimmune serum. Our results provide a baseline for future studies on MC-1R expression in diseased human skin. PMID- 10816670 TI - Effects of ethanol consumption on beta-endorphin levels and natural killer cell activity in rats. PMID- 10816671 TI - Murine dendritic cells express functional delta-type opioid receptors. PMID- 10816672 TI - Expression of corticotropin releasing hormone in malignant melanoma. PMID- 10816673 TI - Depression modulates pruritus perception. A study of pruritus in psoriasis, atopic dermatitis and chronic idiopathic urticaria. PMID- 10816674 TI - alpha-MSH immunomodulation acts via rel/NF-kappa B in cutaneous and ocular melanocytes and in melanoma cells. PMID- 10816675 TI - Microinjection of alpha-MSH followed by UV-irradiation blocks HSP 72 in human keratinocytes. PMID- 10816676 TI - beta-Endorphin binding and regulation of cytokine expression in Langerhans cells. AB - Neuropeptides and neurohormones have been shown to be able to regulate cutaneous immune reactions. Binding of beta-endorphin (beta-end) on epidermal Langerhans cells (LC) and effects of beta-end on cytokine expression were examined. Biotinylated beta-end bound to the mouse LC-like cell line, XS52, and the binding was replaced with intact beta-end but not with substance P. beta-End augmented secretion of IL-1 beta and IL-10 from XS52 cells were induced by a combination of LPS and GM-CSF. Induction of TNF alpha was suppressed by beta-end. The regulation of cytokine expression was confirmed in fresh LC by RT-PCR. These results suggest that beta-end is a regulator of skin immune function. PMID- 10816677 TI - alpha-MSH reduces vasculitis in the local Shwartzman reaction. PMID- 10816678 TI - Implications of the phenotype of POMC deficiency for the role of POMC-derived peptides in skin physiology. PMID- 10816679 TI - Serotonin in human allergic contact dermatitis. PMID- 10816680 TI - Differential temporal and spatial expression of POMC mRNA and of the production of POMC peptides during the murine hair cycle. PMID- 10816681 TI - Effective treatment of pruritus with naltrexone, an orally active opiate antagonist. PMID- 10816682 TI - Developmentally regulated expression of alpha-MSH and MC-1 receptor in C57BL/6 mouse skin suggests functions beyond pigmentation. PMID- 10816683 TI - Plasma beta-endorphin concentrations during natural and artificially induced winter hair growth in mink (Mustela vison). PMID- 10816684 TI - Expression of proopiomelanocortin peptides and prohormone convertases by human dermal microvascular endothelial cells. PMID- 10816685 TI - ACTH production in C57BL/6 mouse skin. PMID- 10816686 TI - Coexpression of nitric oxide synthase and POMC peptides in the dystrophic C57BL/6J mouse. PMID- 10816687 TI - Ligand binding profile and effects of melanin-concentrating hormone on fish and mammalian skin cells. PMID- 10816688 TI - Occurrence of four MSHs in dogfish POMC and their immunomodulating effects. PMID- 10816689 TI - Specific binding sites for beta-endorphin on keratinocytes. PMID- 10816690 TI - Skin POMC peptides. Their binding affinities and activation of the human MC1 receptor. PMID- 10816691 TI - The expression of alpha-MSH by melanocytes is reduced in vitiligo. PMID- 10816692 TI - alpha-MSH inhibits lipopolysaccharide induced nitric oxide production in B16 mouse melanoma cells. PMID- 10816693 TI - The impact of culture on technology. PMID- 10816694 TI - Female sterilisation. PMID- 10816695 TI - The "lost generation": IT education for healthcare professionals. PMID- 10816696 TI - The communication revolution: winners and losers. PMID- 10816697 TI - Technology in patient care. PMID- 10816698 TI - Surgery in the information age. AB - Currently, surgical procedures involve direct data flows of sensation and mechanical output. As information technology progresses, surgery will change to a system of electronic data flows, with technical, ethical, and training implications. PMID- 10816699 TI - The potential impact of home telecare on clinical practice. AB - Home telecare, in which the health status of patients at home is monitored remotely, has the potential to improve care and reduce costs. Its widespread implementation would require fundamental changes in the healthcare system. PMID- 10816700 TI - Health online: the future isn't what it used to be. AB - Over the next 10 years, the healthcare system will change to focus more on preventive medicine and healthcare in the home, with fewer doctors and a new class of home healthcare providers. Healthcare professionals need to debate how best to manage these changes. PMID- 10816701 TI - General practice and the new technologies. PMID- 10816702 TI - The stage is set for the diffusion of positron emission tomography (PET) in oncology. PMID- 10816703 TI - Positron emission tomography (PET): experience with a large-field-of-view three dimensional PET scanner. AB - Positron emission tomography (PET) using fluorine-18 fluorodeoxyglucose (FDG) is an accurate technique for staging and therapeutic monitoring in oncology. We evaluated our use of FDG PET in an oncology centre after our first 2500 studies, and summarise our experience of PET for the major referral indications. Optimised for clinical service, PET offers lower scanning costs and therefore improved cost effectiveness. PMID- 10816704 TI - Filmless radiology: a dream becomes reality. PMID- 10816705 TI - Culture shock. Molecular methods for diagnosis of infectious diseases. AB - Nucleic acid detection methods, such as polymerase chain reaction (PCR), can often detect specific microbial pathogens, virulence markers and antimicrobial resistance genes more rapidly and with greater sensitivity and specificity than culture and conventional identification and susceptibility testing. Multiplex PCR can detect multiple genes in a single assay; this capability will be greatly extended by new techniques such as the DNA chip. However, limitations and pitfalls of nucleic detection methods remain. PMID- 10816706 TI - Electronic prescribing and computer-assisted decision support systems. PMID- 10816707 TI - The Clinical Information Access Project. PMID- 10816708 TI - Providing evidence-based answers to clinical questions. A pilot information service for general practitioners. AB - OBJECTIVE: To pilot a clinical information service for general practitioners. METHODS: A representative sample of 31 GPs was invited to submit clinical questions to a local academic department of general practice. Their views on the service and the usefulness of the information were obtained by telephone interview. RESULTS: Over one month, nine GPs (29% of the sample, 45% of those stating an interest), submitted 20 enquiries comprising 45 discrete clinical questions. The median time to search for evidence, appraise it and write answers to each enquiry was 2.5 hours (range, 1.0-7.4 hours). The median interval between receipt of questions and dispatch of answers was 3 days (range, 1-12 days). CONCLUSIONS: The GPs found the answers useful in clinical decision making; in four out of 20 cases patient management was altered. PMID- 10816709 TI - Health technology assessment in Australia. PMID- 10816710 TI - Assessing new health technologies: lessons to be learned from drugs. AB - Balancing the interests of producers, consumers and purchasers (governments) increases the complexity of evidence-based evaluation of non-drug health technologies. PMID- 10816711 TI - "Picking winners": assessing new health technology. AB - Are evidence-based approaches ready for health technology? PMID- 10816712 TI - Telehealth: opportunities and liabilities. PMID- 10816713 TI - Medicolegal risk in telemedicine: risk control in teleradiology. AB - Teleradiology, the largest single application of telemedicine in Australia, has the potential to improve healthcare access, delivery and standards, but also raise complex new legal and ethical issues. In the absence of any cases of alleged negligence in teleradiology before the courts, a hypothetical case reveals some of the medicolegal risks. These may be minimised by measures including adherence to the Royal Australian and New Zealand College of Radiologists' guidelines on teleradiology. PMID- 10816714 TI - Digital camera and the Internet: bringing the patient home. PMID- 10816715 TI - [Central nervous system demyelinating disease following hepatitis B vaccination with GenHevac B. Review of ten years of spontaneous notifications (1989-1998)]. AB - OBJECTIVES: To describe and analyze spontaneous reports of central nervous system (CNS) demyelinating disease including multiple sclerosis, following vaccination with GenHevac B vaccine, from 1989 to December 31, 1998. METHODS: Descriptive analysis of adverse event reports in the vaccinated population, including the number of cases of CNS demyelinating disease, their frequencies, their dates of onset in relation to dates of report and their distribution according to age, sex and the number of injections. A Kaplan-Meier curve was used to analyze the time period between the last dose of vaccine and the onset of CNS demyelinating disease. RESULTS: Overall, 187 cases of CNS demyelinating disease were spontaneously reported, (0.54 reports per 100,000 doses of GenHevac B distributed). The average time period between the occurring date of onset of the disease and its subsequent report was 24 months. The average age of onset was 31.7 years old and 73% of cases were women. The time between the last dose of vaccine and the onset of disease was regularly distributed from 1 day to 5 years (median: 60 days). CONCLUSION: These results, together with available clinical, epidemiological data regarding multiple sclerosis, do not suggest a causal relationship between CNS demyelinating disease and vaccination with GenHevac B. PMID- 10816716 TI - [Description of long-term stays, based on PMSI data, in hospital specializing in short-term stays]. AB - OBJECTIVE: In a context of organization of care where the budget of hospitals is a function of the number and of the severity of the in-patients and not of the duration of stay, stays of long duration (SLD) in short-stay hospitals represent a problem of both medical and administrative management. To identify the characteristics of long-duration stays. METHODS: The data used in this retrospective study were drawn from the Standardized Discharge Summaries of the year 1997 of a University Hospital of the Paris area (France). A stay of long duration (SLD) was defined in an arbitrary way as a stay exceeding 30 days. The stays of long duration (> 30 days) were subdivided in "long stays" (from 31 to 60 days) and "very long stays" (more than 60 days). RESULTS: The SLDs represent 3.7% of the discharge summaries of our hospital, among them, 40% are medical DRGs and 60% surgical DRGs. The patients in SLD more often come from other structures of care than the patients having a short duration stay of (SDS) coming mainly from their residence and were also hospitalised in several different units during their stay. Patients having a long stay were more often classified in DRGs outside the principal activity of this hospital (i.e. cardiovascular diseases). CONCLUSION: This first approach suggests that a set of simple descriptive variables (pre-existing and acquired co-morbidity, admission in surgical ward, multi-unit stay...) makes it possible to identify the patients likely to have a long duration stay. Simple variables added to the current hospital minimum medical record would make it possible to consider a predictive approach. PMID- 10816717 TI - [Epidermoid carcinoma developing in a chronic anal fistula]. AB - BACKGROUND: Malignant degeneration of anal fistula is a rare occurrence in the course of a common disease. Abdominoperineal resection offers the only hope of cure. CASE REPORT: A 41-year-old patient who was operated 14 years earlier for anal fistula presented recurrence treated by fistulectomy. Histology revealed a well-differentiated squamous-cell carcinoma. Radiotherapy (45 Gy) was added postoperatively. Five months later, local recurrence was treated by chemotherapy and abdominoperineal resection. Two months later the patient died from cachexia with local recurrence. DISCUSSION: Biopsies of anal fistula tract and histology of all fistulectomies are required for early diagnosis of possible carcinoma. Abdominoperineal resection offers the only hope of cure. PMID- 10816718 TI - [Serum lipoperoxide in a pregnant woman and in term and premature neonates]. PMID- 10816719 TI - [Acinetobacter baumannii community-acquired pneumonia in a patient with HIV infection]. PMID- 10816720 TI - [Rapid selection that rectifies a diagnosis of arteritis!]. PMID- 10816721 TI - [Effect of a converting enzyme inhibitor, ramipril, on cardiovascular events in high-risk patients]. PMID- 10816722 TI - [Plea for interdisciplinary dimension in medicine. The example of consent]. AB - A philosophical and legal analysis of consent in the patient-doctor relationship leads to operational limitations. Getting over these limits requires a transdiciplinary approach aimed at understanding the living world and modeling its evolution. This approach rests on recognition of different philosophical, legal, sociological, economical, and historical reality levels, permitting a new non-exclusive role of consent in medicine, i.e. the respect of human dignity. PMID- 10816723 TI - [Inhibiting disease progression. Facts, promises and questions]. PMID- 10816724 TI - [Growth factors. Role in the progression of renal lesions]. AB - FROM PATHOPHYSIOLOGY TO THERAPEUTICS: Nephrologists are faced with the continuing problem of helping patients avoid the onset or retard the development of end stage renal failure. Despite the treatments available, the risk is still high for patients and the cost a heavy burden for the public health budget. These facts underline the importance of a detailed understanding of the mechanisms leading to the destruction of renal parenchyma in order to develop therapeutic strategies capable of slowing the inevitable progression of kidney lesions. GROWTH FACTORS: It is currently recognized that a major reduction in the number of functional nephrons, whatever the initial cause, leads in itself to a progressive deterioration of healthy nephrons and finally to complete destruction of the kidney. The underlying mechanisms remain largely unknown. One possible mechanism would involve an overexpression of several growth factors in the damaged renal parenchyma. We present in this review experimental data obtained with various approaches, including pharmacological and/or dietetic modulations and the establishment of transgenic mouse lines, to demonstrate the key role played by growth factors in the progression of renal lesions. The pathways followed by these growth factors in the process of renal destruction as well as certain elements leading to their overexpression are also discussed. PMID- 10816725 TI - [The renin-angiotensin system and progression of kidney disease]. AB - CONSTRICTIVE ACTIVITY OF THE RAS AND DEVELOPMENT OF RENAL DISEASES: Several studies have suggested that the constitutive level of activity of the renin angiotensin system (RAS), and especially of angiotensin converting enzyme (ACE) which plays an important role in the kidney in determining intrarenal angiotensin and kinin concentrations, is genetically determined and linked to the risk of developing several vascular diseases, including diabetic nephropathy, and to the risk of renal function deterioration in glomerular diseases of several origins. INHIBITION OF THE RAS AND PROGRESSION OF RENAL DISEASES: Large controlled clinical trials have shown over the past years that inhibition of ACE has a beneficial effect and protects against degradation of renal function in type I diabetes with microalbuminuria and also in renal diseases of several origins. PATHOPHYSIOLOGY: All these observations taken together suggest that activation of the RAS, which is necessary in certain circumstances to maintain glomerular filtration and tissue perfusion, can have a long-term deleterious effect on the heart, vessels, and kidneys, especially through glomerular hypertension which can lead to glomerulosclerosis. They support the use of ACE inhibitors, within the indications deduced from analyses of the large clinical trials, for protecting renal function in kidney diseases. PMID- 10816726 TI - [Role of matrix metalloproteinases in the progression of renal lesions]. AB - METALLOPROTEINASES: The progression to end-stage kidney disease is accompanied by accumulation of extracellular matrix (ECM) proteins. The degradation of ECM proteins occurs by the action of proteases, notably the matrix metalloproteinases (MMP). MMP can be classified into four major groups, such as interstitial collagenases, gelatinases, stromelysins, and membrane-type (MT)-MMP. MMP are also involved in the regulation of cell proliferation and possibly of apoptosis. In the kidney, MMP are synthesized by intrinsic glomerular cells and tubular epithelial cells. EXPERIMENTAL DATA: MMP and TIMP expression is well studied in a variety of kidney disorders, particularly in diabetes mellitus and in experimental glomerulonephritis. In diabetes mellitus, an increased plasma concentration of MMP-9 was found to represent the earliest marker of diabetic kidney disease. In anti-Thy 1.1 nephritis, a rat model of mesangial proliferative glomerulonephritis, an increase in proliferation of mesangial cells is associated with augmented expression of MMP-2 and accumulation of extracellular matrix proteins. These inflammatory features can be attenuated by a synthetic MMP inhibitor. ABNORMAL MMP AND TIMP REGULATION: The hypothesis that MMP play an important role in the progression of nephropathies is mainly based on the well known ability of MMP to degrade ECM components, on the association of altered MMP expression in a large number of kidney diseases, and on the ability of MMP-2 to induce or to sustain an inflammatory mesangial cell phenotype. However, since there are only few interventional or functional studies, uncertainty persists as to whether abnormal regulation of MMP and TIMP represents a cause or simply an effect of the respective renal diseases. A DUAL ROLE: In nephropathies, MMP appear to play a dual role as antifibrotic enzymes and as proinflammatory mediators. The exact biologic function in a given renal disorder may depend upon the level of net MMP activity and on the acuteness or the chronicity of the respective disease. PMID- 10816727 TI - Interaction of Saccharomyces cerevisiae with gold: toxicity and accumulation. AB - This paper examines the effects of ionic gold on Saccharomyces cerevisiae, as determined by long-term (growth in gold-containing media) and short-term interactions (H+ efflux activity). An increasing gold concentration inhibited growth and at < 0.2 mM Au, growth was not observed. Transmission electron microscopy revealed no differences in ultrastructure but fine electron dense particles were observed in unstained preparations from gold-containing medium. After glucose addition (to 10 mM) to starved suspensions of S. cerevisiae, glucose-dependent reduction of external pH occurred as the cells extruded protons. In the presence of increasing gold concentrations, the lag time before proton extrusion did not change but the rate and duration decreased significantly with a marked influence on proton efflux rate being observed at < or = 10 microM. Extension of preincubation time of yeast cells in gold-containing medium resulted in a decreasing proton efflux rate and colloidal phase formation in the cell suspensions, the time between gold addition and the beginning of colloidal phase formation depending on the gold concentration used. Both Ca and Mg enhanced the inhibitory effect of gold on the yeast cells with Ca showing a stronger inhibitory effect than Mg. PMID- 10816728 TI - Iron uptake by the yeast Pichia guilliermondii. Flavinogenesis and reductive iron assimilation are co-regulated processes. AB - Pichia guilliermondii cells overproduce riboflavin (vitamin B2) in responce to iron deprivation. The increase in ferrireductase activity in iron-starved P. guilliermondii cells correlated with the increase in flavin excretion. As in Saccharomyces cerevisiae, a typical b-type cytochrome spectrum was associated with the plasma membrane fraction of P. guilliermondii and the cell ferrireductase activity was strongly inhibited by diphenylene-iodonium, an inhibitor of flavoproteins, in both yeasts. Mutants of P. guilliermondii with increased ferrireductase activity were selected for further investigation of the relationship between iron reduction/uptake and flavin production. The obtained mutation has been called hit (high iron transport). A hit mutant with a single recessive mutation showed the following phenotype: high ferrireductase activity, increased rate of iron uptake and elevated flavinogenic activity. Cu(II) (50 microns) strongly inhibited the growth of the hit mutant compared to the wild type. The mutant cells grown in copper-supplemented medium (5-25 microns) showed an increase of the ferrireductase activity (up to 2-3 fold). The copper content of the mutant cells grown under these conditions was also higher (1.5-2 fold) than that of the wild-type. The role of the HIT gene of P. guillermondii in the regulation of iron, copper and flavin metabolisms is discussed. PMID- 10816729 TI - Identification of a fungal triacetylfusarinine C siderophore transport gene (TAF1) in Saccharomyces cerevisiae as a member of the major facilitator superfamily. AB - Transport proteins of microorganisms may either belong to the ATP-binding cassette (ABC) superfamily or to the major facilitator (MFS)-superfamily. MFS transporters are single-polypeptide membrane transporters that transport small molecules via uniport, symport or antiport mechanisms in response to a chemiosmotic gradient. Although Saccharomyces cerevisiae is a non-siderophore producer, various bacterial and fungal siderophores can be utilized as an iron source. From yeast genome sequencing data six genes of the unknown major facilitator (UMF) family were known of which YEL065w Sce was recently identified as a transporter for the bacterial siderophore ferrioxamine B (Sit1p). The present investigation shows that another UMF gene, YHL047c Sce, encodes a transporter for the fungal siderophore triacetylfusarinine C. The gene YHL047c Sce (designated TAF1) was disrupted using the kanMX disruption module in a fet3 background (strain DEY 1394 delta fet3), possessing a defect in the high affinity ferrous iron transport. Growth promotion assays and transport experiments with 55Fe-labelled triacetylfusarinine C showed a complete loss of iron utilization and uptake in the disrupted strain, indicating that TAF1 is the gene for the fungal triacetylfusarinine transport in Saccharomyces cerevisiae and possibly in other siderophore producing fungi. PMID- 10816730 TI - Effect of some heavy metal ions on copper-induced metallothionein synthesis in the yeast Saccharomyces cerevisiae. AB - Copper-induced metallothionein (MT) synthesis in Saccharomyces cerevisiae was investigated in order to associate this exclusively with Cu2+ in vivo, when cultured in nutrient medium containing other heavy metal ions. Expression of the CUP1 promoter/lacZ fusion gene was inhibited by all heavy metal ions tested, especially Cd2+ and Mn2+. By adding Cd2+ and Mn2+ at 10 microM concentration, the beta-galactosidase activity decreased by about 80% and 50% of the maximum induction observed with 1 mM CuSO4, respectively. Furthermore, cell growth was markedly inhibited by combinations of 1 mM-Cu2+ and 1 microM-Cd2+. Therefore, the yeast S. cerevisiae could not rely on MT synthesis as one of the copper resistance mechanisms, when grown in a Cd2+ environment. In contrast, the presence of Mn2+ in the nutrient medium showed alleviation rather than growth inhibition by high concentrations of Cu2+. The recovery from growth inhibition by Mn2+ was due to decreased Cu2+ accumulation. Inhibitory concentrations of Co2+, Ni2+ and Zn2+ on expression of the CUP1p/lacZ fusion gene were at least one order of magnitude higher than that of Cd2+ and Mn2+. These results are discussed in relation to Cu2+ transport and Cu-induced MT synthesis in the copper-resistance mechanism of the yeast S. cerevisiae. PMID- 10816732 TI - Structure of the pyoverdin PVD 2908--a new pyoverdin from Pseudomonas sp. 2908. AB - An unknown siderophore (pyoverdin) was isolated from the strain Pseudomonas sp. 2908. The structure of the pyoverdin--called PVD 2908--was elucidated by spectroscopic methods and degradation studies. Some other siderophores were identified by LC/ESI-MS-screening based on the knowledge of PVD 2908. PMID- 10816731 TI - Transport and utilization of rhizoferrin bound iron in Mycobacterium smegmatis. AB - Transport and metabolization of iron bound to the fungal siderophore rhizoferrin was analyzed by transport kinetics, Mossbauer and EPR spectroscopy. Saturation kinetics (vmax = 24.4 pmol/(mg min), K(m) = 64.4 microM) and energy dependence excluded diffusion and provided evidence for a rhizoferrin transport system in M. smegmatis. Based on the spectroscopic techniques indications for intracellular presence of the ferric rhizoferrin complex were found. This feature could be of practical importance in the search of novel drugs for the treatment of mycobacterial infections. EPR and Mossbauer spectroscopy revealed different ferritin mineral cores depending on the siderophore iron source. This finding was interpreted in terms of different protein shells, i.e. two types of ferritins. PMID- 10816733 TI - Revised structures of the pyoverdins from Pseudomonas putida CFBP 2461 and from Pseudomonas fluorescens CFBP 2392. AB - Several suggestions for structures of the siderophores (pyoverdins) from Pseudomonas spp. can be found in the literature which are based on a FAB mass spectrometric analysis only. Availability of two original strains of two Pseudomonas spp. allowed to re-investigate the structure of their pyoverdins. In both cases the amino acid sequence had to be corrected. In addition, D- and L amino acids could be identified and located in the peptide chain. The knowledge of the correct structures is important in view of an ongoing study to establish relationships between the nature of the peptide chains of pyoverdins and their recognition by outer membrane proteins. PMID- 10816734 TI - Male reproductive effect of nickel sulphate in mice. AB - Nickel sulphate was administered orally to adult male mice at dose level of 5 and 10 mg/kg body weight (5 days per week) for 35 days. There was no change in body weight. However a significant decrease in absolute and organ-to-body weight ratios of testes, epididymides, seminal vesicles and prostate gland was observed. The sperm abnormality, associated with decrease in sperm motility and sperm count was also observed. Significant alterations in the activities of marker testicular enzymes, viz. sorbitol dehydrogenase (decreases), lactate dehydrogenase (increases) and gamma-glutamyl transpeptidase (increases) associated with histopathological changes in testes, epididymides and seminal vesicles, were also observed. Accumulation of nickel in testes, epididymides and seminal vesicles was also observed. The study reveals that the oral exposure to nickel may affect the histology of testes, epididymides, seminal vesicles and sperms morphology. These testicular and spermatotoxic changes may be responsible for observed male mediated developmental toxic effects. PMID- 10816735 TI - Effect of zinc administration on thyrotropin releasing hormone-stimulated prolactinemia in healthy men. AB - Previous in vitro studies have demonstrated zinc (Zn++) inhibition of basal and of potassium (K+) or thyrotropin-releasing hormone (TRH)-stimulated prolactin (PRL) secretion, in a selective, reversible, and dose-dependent manner. Thus, Zn++ may regulate physiologically pituitary PRL secretion. Furthermore, studies with patients with uremia, cirrhosis or prolactinoma, have shown the coexistence of hypozincemia and hyperprolactinemia and zinc supplementation did not correct hyperprolactinemia in these patients. In normal individuals Zn++ administration produced controversial results on PRL secretion. Here, we investigated whether zinc administration affects TRH-stimulated PRL in healthy men. We found that Zn++ administration does not change the TRH-stimulated PRL. Therefore, in normal conditions, Zn++ does not inhibit TRH-stimulated prolactinemia. In addition, we found that acute increases of blood PRL and TRH do not alter blood Zn++ levels. PMID- 10816736 TI - Cadmium induced lipid peroxidation in rat testes and protection by selenium. AB - The main goal of this study was to investigate the role of cadmium in the promotion of lipid peroxidation in the homogenates of rat testes and the effect of selenium on lipid peroxidation in testes of rats after cadmium injection. Treatment of rats with cadmium resulted in a time- and dose-related accumulation of the metal ions in testes. The concentrations of cadmium, copper, zinc, selenium and iron in the tissues were determined by an atomic absorption spectrophotometer and lipid peroxidation in testes was measured by a spectrophotometer. Cadmium produced enhanced lipid peroxidation in testes. These cadmium-induced changes were accompanied by a significant increase of iron and copper, and a decrease of zinc in testes. Concurrent treatment with selenium and cadmium reduced the cadmium-induced alterations in lipid peroxidation and essential metal levels. Data suggest that lipid peroxidation was associated with cadmium toxicity in testes and that the addition of selenium was found to be effective in attenuation of this effect. PMID- 10816737 TI - Comparison of clastogenic effects of inorganic selenium salts in mice in vivo as related to concentrations and duration of exposure. AB - Inorganic selenium compounds in the diet have been known to protect against cancer in laboratory animals, but were harmful in high concentrations. In the present work, the relative effects of two salts, sodium selenite and sodium selenate, administered to mice in vivo, in different concentrations and durations of exposure, were compared. Aqueous solutions of each salt (7, 14, 21 and 28 mg Kg-1 bw) were fed by gavaging to mice matched in age and sex. The animals were sacrificed at intervals of 6, 12, 18 and 24 h and chromosome preparations were made following the usual schedule of colchicine-hypotonic-fixative-airdrying Giemsa staining. The endpoints screened were chromosomal aberrations (CA) and damaged cells (DC). Both salts affected chromosome structure and spindle formation, sodium selenite being more cytotoxic than sodium selenate. The frequencies of aberrations induced were directly proportional to the concentrations used and duration of exposure. PMID- 10816738 TI - Immunomodulatory effect of selenosemicarbazides and selenium inorganic compounds, distribution in organs after selenium supplementation. AB - Antioxidant properties of selenium producing a protective barrier against free radicals play an important role in numerous metabolic and immunologic processes associated with oxidation-reduction reactions which take place during intracellular digestion of phagocyted bacteria. The aim of our study was to examine the properties of an organic compound of selenium, 4-(o-tolilo) selenosemicarbazide of p-chlorobenzoic acid in terms of its retention in organs, effect on erythropoesis and phagocytic abilities of neutrophiles as well as antioxidant properties in neutrophiles tested with NBT test. This compound as well as inorganic sodium selenate was given to Swiss mice at the dose of 10(-3) g Se/kg for the period of 10 days. The concentrations of selenium in livers of mice treated with sodium selenate and selenosemicarbazide were found to be higher than in controls (18.7 micrograms lg-1 and 23.2 micrograms lg-1 vs. 12 micrograms lg 1, respectively). Analysis of blood cells count has shown a significant decrease in neutrophile levels in both groups treated with selenium. The influence of selenium compounds on phagocytosis and especially NBT test has been determined (3.8% of positive cells in the controls vs. 2.2% and 0.9% in the groups treated with sodium selenate and selenosemicarbazide, respectively). Our preliminary investigations suggest that selenosemicarbazides are biologically active compounds and can modify neutrophile functions. PMID- 10816739 TI - Effect of lithium augmentation on the trace elemental profile in diabetic rats. AB - Energy dispersive X-ray fluorescence technique was employed to study the interactions among various elements, viz.: K, Zn, Br, Fe, Cu, Br & Rb in 4 groups of rats viz. control-GI, diabetic-GII (diabetes induced by i.p. administration of alloxan monohydrate at a dose level of 150 mg/kg b.w; single injection), lithium treated-GIII (lithium administered as Li2 Co3 at a dose level of 1.1 g of Li2 Co3/kg animal diet; free access; serum lithium levels--0.5-1.2 mEq/L) and lithium + diabetic rats-GIV. The different treatments continued for a total duration of 1 month. The K contents were found to be significantly lowered in all the treatment groups which was maximum (28%) in lithium treated diabetic rats. Depression in the levels of Rb was noticed in lithium treated and lithium treated diabetic (G III and G-IV) groups. However, the levels (Rb) remained unaltered in diabetic (G II) group. Interestingly, a significant decline was observed in Fe levels in G III following lithium administration but the levels remained unchanged in G-IV with lithium administration to diabetic rats. On the other hand, lithium treatment to normal (G-III) and diabetic (G-IV) rats indicated statistically significant elevation in the levels of Cu and Br. However, diabetic (G-II) rats did not indicate any elevation in the levels of these two elements. Interestingly, the concentrations of Zn were found to be significantly elevated in all the treatment groups, which was maximum (37%) in G-III (lithium) group. A comparison of various elements from lithium treated diabetic group G-IV with the corresponding elements from the diabetic group G-II, implied a significant depression in K and Rb levels and a significant elevation in the levels of Br. PMID- 10816740 TI - [Use of megazol for the treatment of trypanosomiasis]. PMID- 10816741 TI - [Melarsoprol]. PMID- 10816742 TI - [Chad: mitigated results of a fixed strategy]. PMID- 10816743 TI - [Blue medical libraries: experience in the district health centers]. PMID- 10816744 TI - [From tropical medicine ... to international health]. PMID- 10816745 TI - [Sleeping sickness: danger in indifference!]. PMID- 10816746 TI - [Efficacy of permethrin-impregnated Olyset Net mosquito nets in a zone with pyrethroid resistant vectors. I--Entomologic evaluation]. AB - The efficacy of permethrin-treated Olyset Net mosquito nets on malaria transmission and morbidity was studied in Kafine, a village located in the savanna region of the Cote d'Ivoire in Africa. After collecting sociodemographic, entomological, and parasitological data, bednets were distributed first in the southern half of the village and then in the whole village. Throughout the study period, mosquito specimens were captured on the skin of inhabitants at four points in the village between 6 PM and 6 AM both inside (but outside bednets) and outside houses. Prior to distribution of bednets, the mean biting rate (MBR) by Anopheles gambiae was 77.4 bites per man per night (b/m/n). The mean parturity rate (MPR) was 40.6 p. 100, the sporozootic index (SI) was 0.99 p. 100, and the mean entomological inoculation rate (MEIR) was 0.7 infectious bites per man per night (b+/m/n). Six months after distribution of bednets in the southern half of the village, MBR was 80.2 b/m/n, MPR was 32 p. 100, SI was 1.8 p. 100, and MEIR was 0.83 b+/m/n. After extending distribution to the whole village, data from November 1996 to July 1997 were as follows: MBR, 67.8 b/m/n; MPR, 20.1 p. 100; SI, 0.65 p. 100; and MEIR, 0.66 p. 100. From August 1977 to July 1998, data were as follows: MBR, 102.6 b/m/n; MPR, 26.2 p. 100; SI, 1.15 p. 100; and MEIR, 0.74 b+/m/n. Comparative analysis of these data showed that use of bednets had no effect on the bite or entomological inoculation rate. This is in agreement with the documented resistance of vectors in the study zone to permethrin. Despite the known stimulation/repulsion effect of permethrin, use of treated bednets had no real impact on transmission. This inefficacy could be related to the high prevalence (80 p. 100) of the Kdr gene (responsible for resistance) in the savanna form of Anopheles gambiae. PMID- 10816747 TI - [Efficacy of permethrin-impregnated Olyset Net mosquito nets in a zone with pyrethroid resistance vectors. II. Parasitic and clinical evaluation]. AB - The efficacy of permethrin-treated Olyset Net mosquito nets in preventing transmission and morbidity of malaria was studied in Kafine, a village in the rice-growing region of the Cote d'Ivoire. Entomological data demonstrated that permethrin-treated mosquito nets failed to reduce transmission of malaria. Laboratory tests showed that Anopheles gambiae s.s. in Kafine were resistant to permethrin and other pyrethroids. Study included a cohort of 163 children under five years of age who did or did not use mosquito nets. The number of patients seeking treatment for malaria attacks with fever was recorded weekly. No difference was found between the user and non-user groups with regard to the percentage of children exhibiting Plasmodium flaciparum trophozoites or gametocytes or to the mean parasite load. However the rate of high density parasitemia and malaria attacks was twice as high in the non-user group. This difference between users and non-users of mosquito nets cannot be explained only as the result of a physical barrier against bites. In areas where mosquitoes are resistant to pyrethroids, tests conducted in experimental huts have shown that impregnated mosquito nets reduce host/vector contact. However further study will be need to evaluate the overall impact of impregnated nets on malaria in the areas where mosquitoes are resistant to insecticides. PMID- 10816748 TI - [Malaria in expatriates in Abidjan]. AB - This study was carried out in 1998 to assess attitudes and practices with regard to prophylaxis for malaria in expatriate families with children attending French and international schools in Abidjan, Cote d'Ivoire. Data was obtained for a total of 4990 subjects of various ages. Forty point five percent of families reported use of drug prophylaxis. Use of drug prophylaxis decreased with age (69.2 p. 100 up to 1 year versus 34.5 p. 100 after 15 years). The proportion of subjects using bednets was 7.5 p. 100. Some subjects reported use of ineffective or dangerous substances such as pyrimethamin alone (17.5 p. 100) or amodiaquin. The incidence of reported malarial attacks increased in function of the length of stay in the country (50 p. 100 after 15 years). The most effective method of drug prophylaxis appeared to be a chloroquin and proguanil combination which was used by 25.4 p. 100 of subjects. The incidence of malarial attacks in subjects using this combination was 20.8 p. 100 as compared to 43.2 p. 100 in subjects using no drug prophylaxis. Forty-seven percent of the study population reported use of other preventive measures but none correlated with a decrease in the number of malaria attacks. Information concerning preventive measures is readily available but sometimes contradictory. Information provided locally seems to be the most effective but not always the best adapted. To increase the awareness among expatriates, it will be necessary to provide more accurate information about effective preventive measures especially regarding bed nets and drugs. The suggested optimal duration of chemoprophylaxis is unclear but this study shows that there is no difference between those that do or do not use it when the length of stay exceeds ten years. PMID- 10816749 TI - [Prevalence of HIV infection in patients with pellagra and pellagra-like erythemas]. AB - Pellagra is a systemic disorder caused by severe niacin deficiency. While uncommon in Europe and North America, pellagra and pellagra-like erythema are frequently encountered in undernourished adults in poor African countries. The purpose of this three-year prospective study was to determine the prevalence of HIV infection in patients with pellagra. Between 1996 and 1998, all documented cases of pellagra and pellagra-like erythema diagnosed in the Dermatology Department and Internal Medicine Department of the Teaching Hospital in Lome, Togo were included. Patients underwent screening tests for HIV infection. During the study period, pellagra or pellagra-like erythema was diagnosed in a total of 108 patients (59 women and 49 men) with a mean age of 41 +/- 3.5 years (range, 18 to 68 years). Serology tests for HIV were positive in 6 of these patients (5.5 p. 100; mean age 35 years). In four asymptomatic patients with no opportunistic infection, detection of HIV was an incidental discovery. The other two patients had AIDS symptoms. The principal causes of pellagra were malnutrition (n = 30), alcoholism (n = 15), and combined malnutrition and alcoholism (n = 60). The findings of this study suggest that the incidence of HIV infection in patients with pellagra and pellagra-like erythema is low, i.e., not higher than in the general population. This study also confirms previous etiologic and epidemiological data concerning pellagra in poor countries, i.e., the preponderant role of nutritional deficiency. PMID- 10816750 TI - [A new case of subcutaneous dirofilariasis]. AB - Authors describe another case of subcutaneous dirofilariasis due to Dirofilaria (Nochtiella) repens in France. A 26-year-old woman was infected while on vacation in Cap d'Agde on the Mediterranean coast. The patient presented with a subcutaneous nodule in the right subclavicular region. Examination of the nodule after surgical excision revealed the presence of a worm identified as an immature female Dirofilaria repens. Dirofilariasis is a rare anthroponotic disease encountered only in the Old World, particularly in Southeastern France including Corsica which has the second highest number of reported cases after Italy. Since man is a dead-end for the parasite, Dirofilaria repens does not mature and hence most human infections present as isolated subcutaneous nodules. Nodules are usually located in areas exposed to bites by the dipteres, i.e. the face (46 p. 100 of cases reported in the world) and the periocular and palpebral region (30 p. 100). Diagnosis is based mainly on morphological examination of the worm after surgical excision. However promising results in diagnosis of ocular and visceral forms of Dirofilaria repens and understanding of helminthiasis have been achieved thanks to progress in immunological techniques, i.e., ELISA and western blot, and DNA analysis based on polymerase chain reaction. PMID- 10816751 TI - [Uterine perforation after an illegal abortion]. AB - This study focuses on problems related to the management of peritonitis following non-medically assisted abortions in developing countries. Between January 1, 1997 and December 31, 1998, four cases of peritonitis due to perforation of the uterus occurred in a consecutive series of 101 women treated following non-medically assisted abortions at the Gynecology and Obstetrics Clinic of the University Hospital Center in Dakar, Senegal. Abortions were performed by untrained persons using dangerous instruments (wood or metal probes) for prices ranging from 5000 to 30,000 CFA francs. The mean interval between abortion and hospitalization was seven days. All patients presented in poor condition with low-grade symptoms of peritonitis. In 3 of 4 cases, the site of perforation was located in the isthmus (anterior, posterior, and lateral). Extensive necrotic lesions required hysterectomy. Postoperative complications occurred in 3 cases including parietal infection in one case, repeat peritonitis requiring re-operation in one case, and fatal iliomesenteric infarction in one case. Prevention could best be achieved by reducing unwanted pregnancies by better sex education and access to contraceptive techniques. PMID- 10816752 TI - [Scytalidium dimidiatum pseudodermatophyte, agent of superficial mycoses and phaehyphomycosis]. AB - Scytalidium dimidiatum is a fungus found mainly in tropical and subtropical zones. Infection can cause a benign disease closely resembling dermatophytosis. In immunocompromised hosts, Scytalidium dimidiatum can also lead to phaehyphomycosis. Although awareness of these hyphae remains limited in developed countries, their incidence is growing due to increasing immigration and tourism. The rising incidence is well illustrated by three patients who presented onyxis and squamous-like manifestations on the arch of the foot upon returning from trips overseas and in whom various treatments were unsuccessful. In all three cases, culture in non-selective Sabouraud medium identified Scytalidium dimidiatum. These findings underline the need for laboratory testing before undertaking local or systemic treatment of onyxis especially since this pathogen can cause systemic disease. Study of ribosome genes showed that Scytalidium hyalinum is an homologous unpigmented mutant form of Scytalidium dimidatum. No antifungal agent has been effective for management of superficial manifestations and prevention depends mainly on the use of appropriate footwear in endemic areas. PMID- 10816753 TI - [Coexistence of mass hysteria, konzo and HTLV-1 virus in the Democratic Republic of the Congo]. AB - A cross-sectional study was carried out in Pindi located 115 kilometers from Kikwit, Democratic Republic of the Congo to characterize a local school epidemic involving paralysis of the lower extremities, identify risk factors, and establish differential diagnosis with konzo and spastic paralysis related to human T-lymphotropic virus type 1 (HTLV-1). Data was obtained using a qualitative approach based on records, interviews, focus group technique, and neurological examination. Blood tests using the ELISA and western blot tests were performed to detect HTLV-1 and HIV 1 and 2. A total of 41 cases of paralysis were observed between 1994 and 1998. All patients were female and most (n = 28) were between the ages of 16 and 20 at the time of the study. The majority of cases were recorded in 1998 (31 prevalent cases and 16 incidents). Epidemiological data, clinical findings, and laboratory tests suggested that the etiology was mass hysteria with somatic conversion rather than toxic or viral causes in most cases. The psychosocial environment played an important role in the spread of the epidemic. These findings demonstrate the crucial role of the psychosocial environment in the occurrence of mass hysteria and support use of integrated health programs in developing countries. PMID- 10816754 TI - [Tolerance of quinine administered as an intrarectal solution in children in French-speaking Africa]. AB - Pharmacokinetic studies have documented the efficacy of intrarectal quinine (IRQ) for treatment of childhood malaria. As a result, this technique has gradually supplanted intramuscular injection of quinine (IMQ), a common source of complications in children. To assess the safety of this technique, outcome was routinely monitored from 1987 to 1997. This report presents immediate tolerance observed in clinical studies involving 915 children as well as two surveys conducted in Niamey, Niger on 2764 children in 1995 and on 714 children in 1997. In the second study tolerance of IRQ (n = 364) and IMQ (n = 350) was compared. Long-term tolerance was evaluated for 11 years in a single location, i.e. Morondava, Madagascar. In 1995 IRQ accounted for 19.6 p. 100 of antimalarial treatments prescribed in Niamey. In 1997, IRQ accounted for 34.5 p. 100 of quinine prescriptions (excluding infusions) written in medical centers and 65.7 p. 100 in the Pediatric Department B of the hospital in Niamey. Overall tolerance was good, thus confirming previous clinical studies. No major and/or irreversible complication was observed. Early rejection (12.9 p. 100), intestinal transit problems (4.3 p. 100), and watery stools (12.9 p. 100) were the most common problems. In contrast, IMQ led to residual pain (3.1 p. 100), local inflammation (3.1 p. 100), abscess (0.6 p. 100), and lower extremity disability (0.3 p. 100). The precautions for use, e.g. proper dilution, and staff training requirements are reviewed in the discussion. PMID- 10816755 TI - [Dirofilariasis, an emerging and underestimated zoonoses in France]. AB - A total of 71 cases of dirofilariasis due to Dirofilaria repens and two cases of pulmonary dirofilariasis due to Dirofilaria have been reported in France. As a result France ranks second after Italy in the incidence of this emerging zoonoses that has been given little attention (48 p. 100 of cases in the last ten years). Most human infections (80 p. 100) have occurred in Mediterranean areas but in the last ten years a few cases have been reported in Southwestern France and at least one case was documented in a northern area (Alsace). The epidemiological profile of human dirofilariasis in France is consistent with usual data showing highest incidence in adults and no gender preference. More than 50 p. 100 of superficial nodules are located on the head with the most common site being the conjuctiva and orbit (29.4 p. 100). Nodules on the breast and male genitals as well as pulmonary nodules require differential diagnosis to rule out neoplasm. Intra abdominal forms are usually discovered incidentally. Identification is rarely based on detection of a whole adult worm but rather on observation of a nematode segment on histological sections. In most cases it is an unexpected finding. Data concerning canine heartworm in France is available but the extent of infestation is probably underestimated and will require further study. In addition, data on other animal reservoirs should be updated. PMID- 10816756 TI - [Management strategies for endemic goiter in developing countries]. AB - Despite significant progress in the last decades, endemic goiter remains a serious public health problem in the developing world, especially in Africa. Even in countries that have successfully reduced overall incidence to acceptable levels, endemic areas often remain. This persistence is due to the inadequacy of preventive measures and poor follow-up of control programs. The main etiologic factor in endemic goiter is inadequate dietary intake of iodine. This commonly occurs in communities depending exclusively on local produce grown on iodine-poor land, especially in mountain areas. Endemic goiter is epidemiologically associated with cretinism, deaf-mutism, and mental retardation. Even mild iodine deficiency leads to clinical hypothyroidism and moderate myxoedema with significantly reduced intellectual performance. Prevention of endemic goiter depends mainly on increasing the iodine intake of people in endemic areas. When iodine intake reaches the estimated adult minimum requirement (100 to 150 micrograms per day), the prevalence of goiter decreases. Two approaches have been used to increase iodine intake. The first consists of adding iodine to food staples such as table salt. The second consists of medical treatment using agents such as iodized oil. Iodization or iodination of salt is the most widespread and cost-effective method of prevention. Administration of iodized oil has been used only in severely endemic areas and in regions where reliable provision of iodinized salt is prevented by geographical barriers or political factors. However, iodized oil has been helpful in the start-up phase of prevention programs using iodized salt, either as an emergency measure or as a mean of convincing officials of the efficacy of iodine prophylaxis. PMID- 10816757 TI - [A case of Ebola virus hemorrhagic fever in Libreville (Gabon), fatal after evacuation to South Africa]. PMID- 10816758 TI - [Neonatal screening for sickle cell anemia and other hemoglobinopathies in Martinique]. PMID- 10816759 TI - [Acute optic neuromyelitis or Devic syndrome in a six-year old child in Burkina Faso]. PMID- 10816760 TI - Role of nuclear lamins in nuclear segmentation of human neutrophils. AB - Nuclear breakdown leading to the formation of apoptotic bodies has been postulated to involve degradation of nuclear structural proteins, such as lamins A/C and B. Although nuclear segmentation occurs during the maturation of polymorphonuclear leukocytes (neutrophils), its mechanism is not known. We found that human neutrophils have lamin B but lack lamins A/C while mononuclear cells possess all three types of lamin as assessed by immunoblotting. Differentiation of human promyelocytic HL-60 cells into neutrophil-like cells was also accompanied by the down-regulation of lamins A/C but not of lamin B. Moreover, when compared with normal cells, neutrophils with the Pelger-Huet anomaly of nuclear hyposegmentation exhibited significantly lower activity of caspase-6, a lamin A/C-cleaving enzyme. Differentiated HL-60 cells showed higher activity of caspase-6 than that of untreated cells. These observations allow us to speculate that remodeling of nuclear lamins might underlie the mechanism for nuclear segmentation of neutrophils. PMID- 10816761 TI - Bis[(benzo-15-crown-5)-15-yl methyl] pimelate forms ion channels in planar lipid bilayer: a novel model ion channel. AB - Some crown ethers translocate cations across the liposomal membrane either by a carrier mechanism or by forming ion channels. We report formation of ion channels in lipid bilayer membranes by bis[(benzo-15-crown-5)-15-yl methyl] pimelate, a crown ether known to form ion inclusion complexes with alkali metal cations. The channels have characteristic long openings lasting several seconds and a low conductance (4 pS in 500 mM KCl and 2.5 pS in 500 mM NaCl). A model of the crown ether channel formed by stacking of four monomers is proposed. A large database of structural information on crown ethers and their ion inclusion complexes as well as large family of crown ethers with a variety of substitutions in the ring are commercially available. Thus the crown ether channel is an attractive model system to study the role of various chemical moieties in ion conduction which may provide deeper insight into understanding the mechanism(s) of selectivity, ion transport, etc. in biological ion channels. PMID- 10816762 TI - In vitro high resolution proton magnetic resonance study of human cerebellar development during the period from the fetus to childhood. AB - In the development of the human cerebellum, the intracellular metabolites were monitored during the period from the fetus to childhood by in vitro high resolution proton (1H) magnetic resonance (MR) spectroscopy. The spectra from fetus (15-30 post-menstrual weeks; n = 3), infant (1-24 months of age; n = 6) and child (7-14 years of age; n = 5) groups showed resonances from seventeen different metabolites. The level of N-acetylaspartate (NAA), one of the metabolites, was observed in age-dependent increases, two- and three-fold increases for infant and child groups from the NAA of the fetus group, respectively. The rapid increases in the creatine (Cre) level (approximately three-fold) in the fetus and infant groups were observed in the child group (approximately four-fold). Taurine (Tau) was noted at the highest concentration in the fetus group. Slight increases in concentrations of alanine, glutamate, glutamine, and glycine and a significant increase in the concentration of N acetylaspartylglutamate were also noted in the fetus and infant groups. Other metabolite concentrations did not change significantly throughout the studied age groups. These findings indicate that synthesis of metabolites, especially of NAA and Cre, during the development of the cerebellum are closely correlated with mitochondrial energy metabolism, and as such, may reflect mitochondrial integrity in the cerebellum. PMID- 10816763 TI - Effects of chronic physical activity and of ultrasound treatment on bone consolidation. AB - The objective of this study was to investigate the effects of ultrasound treatment and physical exercise on the velocity of bone consolidation and resistance to deformation. We performed osteotomy in the upper third of the right tibia of rats. Physical training consisted of swimming 1 h per day with a load of 5% b.w. for 30 days. Therapy with medium-intensity ultrasound was applied daily on the damaged area. Wistar rats were divided into the following groups: osteotomized sedentary animals with no ultrasound treatment (1.OSnUS), osteotomized trained animals with no ultrasound treatment (2.OTnUS), osteotomized sedentary animals with ultrasound treatment (3.OSwUS), and osteotomized trained animals with ultrasound treatment (4.OTwUS). The animals were sacrificed for the following analyses: muscle glycogen, serum alkaline phosphatase at the 5th, 10th, 20th, and 30th days, test of maximum resistance to flexion, rupture flexion and mean tibial rigidity at the 30th day. Muscle glycogen was increased at the 20th day; alkaline phosphatase was elevated at the 5th and 20th days in groups 3.OSwUS and 4.OTwUS, and decreased at the 10th day. Groups 1.OSnUS and 2.OTnUS did not show significant variations. In the mechanical resistance tests, we noted that ultrasound therapy and the association of physical activity used in the present study showed significant differences in bone resistance and bone rigidity after 30 days of treatment. These facts suggest that ultrasound or physical activity, or their combination may accelerate the process of bone tissue repair. PMID- 10816764 TI - The static magnetic field effects on ouabain H3 binding by cancer tissue. AB - Radioactive labeled ouabain was used for estimating the static magnetic field (SMF) induced cell volume changes. Ouabain is a specific inhibitor of Na+/K+ ATPase, and can be used for estimating its quantity--thus giving information about the cell volume changes. Ouabain binding by cancer and normal glandular tissues of breast cancer patients and normal glandular tissues of healthy women was measured after exposure of tissues to SMF 0.2T. SMF exposure led to a decrease of ouabain binding in both normal and cancer tissues when ouabain concentration in the external medium was 10(-9) M, while in the case of higher concentrations of ouabain (10(-7) M, 10(-6) M) an increase of ouabain binding was seen. The normal glandular tissues of healthy women were sensitive to SMF only at the highest concentrations of ouabain used in our experiments. The SMF-induced decrease of binding at low ouabain concentrations was considered as an evidence for the dehydration effect of SMF. It is suggested that the SMF could influence the cancer cell metabolism through cell hydration changes. PMID- 10816765 TI - A theoretical study of the radiological properties and water equivalence of Fricke and polymer gels used for radiation dosimetry. AB - A method is described which determines the radiological equivalence of different materials by comparing their macroscopic photon and electron interaction parameters over the energy range of interest. This method has been applied to Fricke and polymer gels used for radiation dosimetry. Photon and electron dose distributions were calculated using a Monte Carlo technique. 6 MV photon gel depth dose curves were all within 1% of those in water over the depth range studied. 6 MeV electron beam depth dose curves were all within 1 mm of those in water. The results show that for the gels investigated, at megavoltage energies the dominant property determining water equivalence is the electron density of the gel, and therefore future research into gel manufacture for external beam radiotherapy should aim to have the electron density (electrons per unit volume) as close as possible to water. However, care needs to be taken when using gels for dosimetry of photons with incident energy < 100 keV. PMID- 10816766 TI - The inhomogeneous conductivity property of cranial tissues and its representation in numerical head models. AB - The inhomogeneous conductivity of cranial tissues is discussed in this paper. A novel approach for including detailed tissue inhomogeneity is proposed and developed using pseudo-conductivity values which are derived from the limited data available from physiological experiments. Simulation studies of the effects caused by the inclusion of inhomogeneity in the EEG forward problem are also presented in this paper. The results suggest that the accurate representation of the tissue inhomogeneity improves the quality of the computed EEG forward solution. Conclusions about the effects of inhomogeneity on the potential distribution are given. PMID- 10816767 TI - Comparison of effective source-surface distances for electron beams derived from measurements made under different scatter conditions. AB - The purpose of this paper is to investigate the effect of scatter conditions around the chamber on the effective source-surface distance (SSDeff) calculation. Three setups were considered, viz with the chamber: in air, in solid water at the surface, and in solid water at dmax. Ionization measurements for electron beams from a Varian Clinac-210 degrees C were recorded from 100 cm to 120 cm nominal SSD in 5 cm increments, using a parallel-plate Markus chamber. Two electron energies, 6 and 12 MeV, were investigated with a range of electron applicators (Series-III, 10 x 10 to 25 x 25 cm2) and different cerrobend inserts (square and rectangular). The study was undertaken by placing the chamber in air and in solid water, respectively. SSDeff was calculated from the ionisation chamber measurements using the method of Khan (1984). The results are summarised in the table 1. In almost all cases the SSDeff calculated using the in-air data is less. This is likely to be due to a larger proportion of scatter off the applicators reaching the chamber when it is in air than when it is at dmax in the phantom. The results also show the difference is reduced for a larger applicator, probably because the applicator scatter component reaching the in-air measurement point is reduced. PMID- 10816768 TI - A survey of patient dose and image quality factors for CT scanners in Victoria. AB - Image quality and dose parameters have been measured on most makes and models of CT scanners in Victoria. Image quality was assessed using a dose efficiency and imaging performance parameter (Q-value). Factors relating to patient dose were measured. Patient doses using CTDIw in mGy were determined for average size patients for the abdomen procedure. Guidance levels have been recommended for CT procedures in Victoria. PMID- 10816769 TI - Local shielding of high dose rate brachytherapy in an operating theatre. AB - A high-dose rate brachytherapy facility was installed into an established operating theatre by using local shielding in the form of mobile lead screens and by taking advantage of the ease with which staff movements can be controlled in an operating suite. This facility was inexpensive to develop, and has proved clinically efficient and entirely adequate from a radiation safety standpoint. PMID- 10816770 TI - [Tendon transfers to restore hand function following peripheral nerve injury in the arm]. AB - Three patients, a woman aged 33 years, a man aged 42 and a woman aged 66, had a loss of hand function. This was due to denervation of a peripheral nerve, in essence the radial, ulnar and median nerves respectively. In the first two cases this was caused by trauma and in the third by compression neuropathy. The problems were loss of wrist and finger extension (a dropping hand) in the first, loss of intrinsic muscle function and thumb adduction in the second and loss of opposition in the third patient. Tendon transfers were performed by diverting a tendon insertion of a still functional muscle to another tendon to correct the disturbed mechanical balance in the affected upper extremity. After initial immobilisation during several weeks intensive rehabilitation was started. All patients had improvement of range of motion and power in the affected wrist and fingers. In this way a better daily function and resumption of work were accomplished. PMID- 10816771 TI - [Protection from sunlight, particularly for children]. AB - Excessive exposure to ultraviolet radiation (UVR) in childhood has been recognized as an extrinsic risk factor for the development of skin cancer later in life. The risk of squamous cell carcinoma is clearly related to the lifetime accumulated UVR dose, while the risk of melanoma and basal cell carcinoma is increased by intermittent UVR exposure, which is frequently responsible for sunburn. Besides UVR exposure, several intrinsic factors increase the risk of skin cancer, especially the risk of melanoma, such as skin type, the number of normal and dysplastic melanocytic naevi and family history. Responsible sun behaviour in childhood is most important to prevent skin cancer: stay in the shade, especially between 12:00 and 3:00 p.m., wear protective clothing, including a hat and sunglasses, try to build-up natural adaptation of the skin to UVR and apply a sunscreen to UVR exposed skin. There are strong indications that the possible preventive effect of sunscreens on the development of skin cancer is abolished in case of prolonged and intensive UVR exposure, because of a false feeling of safety. The most important measure to prevent the development of melanomas and basal cell carcinomas is probably the prevention of sunburn in childhood. PMID- 10816772 TI - [Physical diagnosis--percussion and palpation of liver]. AB - Percussion and palpation of the liver can produce useful information regarding size, consistency and the surface of the lower liver edge. However, the accuracy of the physical examination of the liver is limited, especially concerning the predictive value of the absence at physical examination of hepatomegaly (> or = 15 cm liver width in the right midclavicular line) for the absence of a truly enlarged liver. Also, the inter-observer variability may be considerable. These limitations of the test may be partly due to a number of pitfalls and misconceptions concerning physical examination of the liver. PMID- 10816773 TI - [Clinical thinking and decision making in practice. A severely ill elderly man with icterus]. AB - A 74-year-old man was admitted to hospital because of jaundice and malaise of several weeks' duration. Five years earlier he had sustained a stroke from which he had recovered almost completely. On physical examination he was overweight and had an enlarged liver. Laboratory values were consistent with cholestasis and hepatitis. An abdominal ultrasound showed multiple nodular abnormalities in the liver consistent with a malignancy. Rapidly developing abnormalities in blood coagulation were thought to be a contraindication to hepatic biopsy. The patient deteriorated and sustained a new stroke. The physicians were convinced that he had cancer and could not be cured. They planned further diagnostic studies, but at the same time made an advance directive for non-resuscitation. Three days after admission the patient was found dead; no consent for an autopsy was obtained. If it is suspected that a patient is suffering from a malignant disease, the malignancy should be demonstrated or excluded as quickly as possible in the least uncomfortable way. Also, the patient and his family should be informed of any restriction on the possibilities of treatment. PMID- 10816774 TI - [Detailed analysis of the societal costs of diabetes mellitus]. AB - OBJECTIVE: Estimation of the societal costs of diabetes and related complications in support of the revision of the diabetes guidelines. DESIGN: Retrospective and descriptive cross-sectional study. METHODS: Relative risks (RRs) of diabetes complications, by age, were determined by comparing patients with and without diabetes. Using existing 1994 General Practitioner registry data on prevalence, on costs of illnesses and on absenteeism, the contribution of diabetes to the costs of other illnesses was estimated on the basis of aetiological fractions calculated with the RRs found. Cost due to absenteeism by diabetes complications were calculated using the friction method. RESULTS: In 1994, the societal cost of diabetes were 1.67 milliard Dutch guilders (1.55-1.87; range determined by applying the 95% confidence intervals of the RRs; in Euro: 758 million (703-848). The costs due to absenteeism from work were almost 0.2 milliard. The medical costs of diabetes were 2.5% of the health care budget. The contribution of diabetes to the medical costs of cardiovascular diseases was 14%. PMID- 10816775 TI - [Scrotal testosterone patches: a good addition to therapeutic options for hypogonadal men]. AB - OBJECTIVE: To report the experience with treatment of hypogonadal men with the scrotal application of testosterone patches. DESIGN: Prospective and descriptive. METHOD: In 12 unselected hypogonadal men (aged 27-58 years; serum concentration of testosterone < 10 nmol/l) who complained of adverse effects of oral androgen treatment, the treatment was changed to scrotal application of testosterone patches (Testoderm). First the androgen substitution was stopped for 6 weeks. Follow-up was 4-5 years, during which regular laboratory tests were carried out, the patients filling in questionnaires regarding their general mood and the frequency of sexual feelings/activities. RESULTS: Testosterone levels reached physiological values and a beneficial effect was observed on general mood and sexual functioning. These effects remained stable throughout the entire period of 4-5 years of study. Skin reactions occurred in 2 men. Dihydrotestosterone levels increased to supraphysiological values. Other clinical, biochemical and haematological parameters did not reveal any detrimental effects. The patch, which should be applied to a dry and shaved scrotal skin, was a satisfying androgen replacement therapy for more than 4 years in 7 out of 12 men. CONCLUSION: In this open study in hypogonadal men with complaints about previous testosterone therapy, scrotal testosterone patches were a useful therapy, leading to the testosterone levels in the physiological range. PMID- 10816776 TI - [Posttraumatic syringomyelia in 2 patients with thoracic spinal cord lesions]. AB - Two patients, men aged 42 and 40 years, developed new neurological symptoms 3 months and 22 years, respectively, after a traumatic high thoracic spinal cord injury. The MRI scan showed a cavity in the central part of the spinal cord, on which the diagnosis of 'posttraumatic syringomyelia' could be based. In one of the patients a syringo-subarachnoidal shunt was created, the other was treated conservatively because of a severe concomitant thoracic kyphosis. Posttraumatic syringomyelia is a potentially life-threathening late complication of spinal cord injury and is characterized by development of new neurological symptoms after a variable time interval. The most typical symptom of non-traumatic syringomyelia, viz. diminution of vital sensitivity without loss of gnostic sensitivity, is not necessarily present in posttraumatic syringomyelia. Surgical treatment of posttraumatic syringomyelia is advocated if there is progressive neurological deterioration, and consists of drainage of the syrinx. PMID- 10816777 TI - [Summary of the 'Cholesterol' guideline (first revision) of the Dutch College of Family Practice]. PMID- 10816778 TI - [Severe Plasmodium falciparum malaria in pregnancy: a threat to mother and child]. PMID- 10816779 TI - Molecular sex identification of stillborn and neonate individuals ("Traufkinder") from the burial site Aegerten. AB - The study reconstructs the sex ratio of 121 stillborn and neonate individuals from the early modern burial site Aegerten, Switzerland. The immature individuals, who were not baptised before death, were buried along the walls of the church. To perform a molecular sex identification, bone samples from the infants were collected from different skeletal elements. Ancient DNA (aDNA) was isolated by a combination of automated phenol/chloroform extraction and precipitation with silica powder. A combination of manuell Chelex extraction and purification kit was also used to perform an extraction. Finally, the aDNA extracts were amplified with a primer system that amplifies a part of the amelogenin gene located on the human sex chromosomes. The morphometrical sex determination of the children suggests a large disproportion of female individuals (about 60%). This finding was compared to PCR-based amplification results. In contrast, the results of the molecular sex identification were a high proportion of male individuals. Looking at these results, it should be noted that the high mortality of male individuals during the last months of pregnancy and during the first month after birth is in accordance with the natural sequence of death also found in recent populations. PMID- 10816780 TI - Evaluation of morphological sex determinations by molecular analyses. AB - The study presents an evaluation of morphological sex determinations of adult skeletal individuals based on traits of the ossa coxae and the cranium. The evaluation criterion was genetic analysis of the amelogenine gene, which represents parts of the X- and the Y-chromosome (Mannucci et al. 1994). The study was carried out on 33 human skeletons from an early modern burial site in Lower Saxony. In this skeletal series, 88% of the morphological sex determinations matched the genetic sex. The percentage of matches was further improved, if only those morphological determinations were taken into account that were classified as unambiguous by a self-evaluation. In the reverse case, a significant number of non-matching determinations (33%) resulted from those cases in which a sex determination still seemed possible but was classified as "ambiguous" in the self evaluation. At least within this skeletal series, no clear connection could be detected between the number of matching results and the presence or absence of the ossa coxae. This might be due to a strong cranial dimorphism within this particular skeletal series. PMID- 10816781 TI - Megaplex DNA typing can provide a strong indication of the authenticity of ancient DNA amplifications by clearly recognizing any possible type of modern contamination. AB - Recent experiments revealed the perfect applicability of megaplex typing by autosomal short tandem repeats (STRs) to degraded DNA. The advantages of megaplex approaches lie in reduced amounts of sample material that are necessary and in remarkable time saving. Furthermore, megaplex typing clearly recognizes possible contaminations and thus has a large potential for indicating authenticity in ancient DNA analysis. This is demonstrated by three examples in which various types of contaminations could clearly be identified as such and even traced back to their origin. This would have been impossible using control samples, due to the sporadic nature of these types of contaminations. PMID- 10816782 TI - Short Tandem Repeat loci analysis in forensic casework. AB - The development of polymerase chain reaction typing method has enhanced the possibilities of genetic identification of biological evidence samples for forensic caseworks. The authors report the use of multiplex amplification reactions associating several short tandem repeat loci. These amplification systems produce reliable results and a sufficient incrimination power to send the cases before the court. PMID- 10816783 TI - STR-genotyping of archaeological human bone: experimental design to improve reproducibility by optimisation of DNA extraction. AB - The analysis of degraded DNA with the help of short tandem repeat loci (STRs) is an important source of information both in forensic casework and in the anthropological context. The reproducibility of STR-genotyping of highly degraded or "ancient" DNA can be reduced by the generation of artifacts during PCR amplification. The frequency and amount of these artifacts--allelic dropout and the generation of shadow bands--are related to the quality and quantity of the extracted DNA amplified in a PCR reaction. Therefore, one important strategy to increase the reproducibility of STR-genotyping of samples containing degraded DNA is the optimisation of the DNA extraction. PMID- 10816784 TI - Ancient DNA-typing approaches for the determination of kinship in a disturbed collective burial site. AB - Several DNA-typing approaches are applied for identification and kinship analysis. Autosomal Short Tandem Repeat (STR) typing produces the genetic fingerprint that is unique to an individual. Y-chromosomal STR typing identifies individuals of the same paternal lineage, and sequence analysis of the hypervariable region of the mitochondrion can identify maternally related individuals. The combined approach of these DNA-typing methods allows the determination of kinship even in complex collective burial situations. In a bronze age collective site, the typing methods were tested for applicability to ancient DNA. For each approach, results were obtained, leading to the conclusion that the determination of kinship is achievable. PMID- 10816785 TI - STR allelic frequencies in a German skeleton collection. AB - Chromosomal DNA was isolated from bones from a German skeleton collection (Goslar, 18th century) and detected by PCR. Nine microsatellite regions were amplified by multiplex reactions using the AmpFlSTR Profiler Plus kit and analysed to obtain their allelic distribution. A statistical evaluation of the results revealed no allelic differentiation between the historic sample and a modern German one at each locus. PMID- 10816786 TI - Ancient DNA fragments longer than 300 bp. AB - It is widely assumed that ancient DNA (aDNA) extracts contain no authentic templates longer than 300 bp. Here we present results which show that fragments of up to 800 bp in length can be reproducibly amplified from aDNA extracts. The amplification involves the short tandem repeat (STR) locus HUMVWA31A. Authentication of the amplified fragments is carried out by measures of expectancy. PMID- 10816787 TI - Paleopathological and molecular evidence of human bone tuberculosis in Iron Age Lithuania. AB - Skeletal remains of two individuals, showing lesions suggestive of bone tuberculosis, from the archaeological sites of Marvele and Sukioniai in Lithuania were analyzed at the DNA level. The diagnosis of bone tuberculosis was confirmed in the remains from Marvele by amplifiying a 245-bp fragment of a repetitive insertion element-like sequence (IS 6110) of Mycobacterium tuberculosis DNA. This is direct evidence for the presence of tuberculosis in Lithuania at the beginning of the first millennium AD. The individual from Sukioniai was found to be tuberculosis-negative. No PCR product was obtained for the 245-bp target sequence or for a smaller 123-bp DNA fragment specific for Mycobacterium tuberculosis. However, amplifiable ancient DNA appeared to be present in the examined specimen as was shown by the results of the DNA-based sex identification, which indicated, consistent with the bone morphology, a male individual. PMID- 10816788 TI - Reconstruction of kinship by fecal DNA analysis of orangutans. AB - Genetic analysis is a useful tool for assigning biological relationships. Thus, it will improve genetic management of wild animal populations and breeding colonies. Kinship analysis will give new insights into the behavior, sociobiology and genetic management of orangutans. In this study, chromosomal DNA from orangutan (Pongo pygmaeus ssp.) was extracted from excrements. Feces samples were screened for up to nine microsatellite markers from related zoo populations of orangutans (Pongo pygmaeus ssp.) kept at the Zoological Garden Berlin and the Zoological Garden Heidelberg, Germany. Family structures are documented in the "International Studybook of the Orangutan" (Perkins 1995) and the "Europaisches Erhaltungszucht Programm 1998" (Becker 1998). To examine whether human short tandem repeat loci (STR) are suitable for the reconstruction of kinship in orangutans, nine STRs, commonly used in forensic studies and the amelogenin system, were amplified in a multiplex-PCR approach (AmpFlSTR Profiler Plus). We were able to show that five of the nine human autosomal STRs in question amplified successfully in orangutans. Thus, we could reconstruct kinship structures of the Berlin and Heidelberg populations. PMID- 10816789 TI - Palaeogenetic analysis of (pre)historic artifacts and its significance for anthropology. AB - The possibility of isolating ancient DNA (aDNA) from all kinds of (pre)historic anthropogenetic artifacts opens new perspectives. This study applies palaeogenetic techniques to three anthropological issues: 1. Palaeodiet. DNA sequences from organic residues in vessels identify Precolumbian Aztec diet. 2. (Pre)historic husbandry and economic structures. aDNA data can reveal the species and the genetic evolutionary stage of animals and plants and show the manner and the extent of their growth, cultivation, or domestication. 3. Production techniques, use, and functionality. Identification of the plant or animal source of an archaeological find can reveal the use or the function of the find. Examples from a Celtic "sausage-end" and an Aztec "eye salve" are given. PMID- 10816790 TI - Trace analysis of endogenous and exogenous biomolecules from archaeological skeleton materials. AB - An analytical procedure was developed for the determination of pharmacologically active substances in archaeological skeleton materials. In comparative model studies, added ("spiked") test biomolecules of varying chemical behaviours were extracted from sample matrices and percentage of the analytes recovered were estimated using gas chromatography/mass spectrometry (GC/MS) or high performance liquid chromatography (HPLC). For the sterols and steroids studied, several organic solvents were appropriate. Extraction yields for the alkaloid nicotine, representing non-endogenous basic agents, were increased by alkalizing with triethylamine or by extracting with a two-phase system consisting of an alkaline aqueous and an organic layer (toluene). The flavonol quercetin was extractable only in an acidic environment. In a screening for native biomolecules using GC/MS, nicotine was identified in individual samples, in addition to lipophilic substances such as the endogenous cholesterol and its degradation products and two phytosterols which may have migrated into the bones from the surrounding soil. PMID- 10816791 TI - Possibilities of extraction and characterization of ancient plasma proteins in archaeological bones. AB - Due to the mineral matrix bone proteins are capable of surviving during centuries after inhumation, but cross-linking with other bone proteins as well as fragmentation and complex reactions with humic acids and microorganisms lead to considerable alterations in molecular weight and structure of these proteins. Our group concentrates on polymorphic plasma proteins which diffuse out of the capillary system into the bone matrix where they adsorb to the mineralic substrate. So far, only little is known about the degradation and alteration of these proteins in fossil bones. It has to be evaluated whether the aged proteins still contain some of the information which renders them a valuable tool for forensic questions and population genetics in recent populations. Therefore we tried by modification of already existing methods to expand plasma protein identification and subtyping into the new field of aged plasma proteins. PMID- 10816792 TI - Molecular genetic analysis of the polymorphic apolipoprotein E in modern and ancient human DNA samples. AB - In this report, methodical bases for the molecular genetic analysis of the three common apolipoprotein E alleles APOE*2, APOE*3 and APOE*4 in DNA isolated from ancient human skeletal remains are described. Considering that ancient DNA target regions for amplification are generally quite small, the detection method is based on short amplification products in the range from 71 bp to 75 bp. The applicability of the modified method for APOE genotyping was examined in modern human DNA samples. PMID- 10816793 TI - Local or nonlocal? A research of strontium isotope ratios of teeth and bones on skeletal remains with artificial deformed skulls. AB - The analysis of strontium isotopic composition of teeth and bone served as an approach to determine nonlocal individuals with artificially deformed skulls in Teuton and Gepid sites. A differing Sr-isotopic composition between tooth and bone from the same skeleton reveals a residence change between early childhood and the last ten years before death. The results show that most Teuton and Gepid individuals investigated are local habitants. PMID- 10816794 TI - Differentiation between occupational and dietary-related intake of barium in the skeleton. AB - On the basis of the reconstruction of general nutritional patterns by trace element data on historic populations, an examination on occupational Ba intakes in the skeleton was carried out for a historical metallurgy population. The concentrations of the trace elements Ba, Sr, and Pb as well as the matrix element Ca were determined by AAS analysis. Their correlations within the population were analysed statistically. Evidence of a correlation between Ba and Sr concentrations and the lack of a correlation between Ba and Pb concentrations is interpreted as evidence for a probably solely dietary-related Ba intake in the skeleton. Ba intake from working in ore smelting plants is therefore unlikely or at least insignificant. These conclusions are supported by analysis results of modern food items from the catchment area of the historical population. A sufficient Ba supply through rye was determined, thereby explaining the concentration of Ba in the bone material of the historic population. PMID- 10816795 TI - Neighbours in different habitats--subsistence and social differentiation in early mediaeval populations of the eastern Swabian Alb. AB - The sites of Neresheim and Kosingen are located in the easternmost part of the Swabian Alb in close proximity, but belong to separate natural units characterized by clearly different ecological properties. By investigating early mediaeval skeletal samples of the populations, two questions were addressed: (1) to what extent can modes of subsistence be explained by the ecological context, and (2) does this affect patterns of nutritional and social differentiation within the populations? Reconstruction of dietary bases by trace element analyses revealed a mixed diet consisting of both crops and animal-derived products at Neresheim, while the nutrition at Kosingen suggests the consumption of food components enriched in dietary calcium with a stronger emphasis on animal products. These results are in accordance with what may be expected based on the options available in the respective habitat. People in Neresheim mostly relied on crop farming in the fertile areas of their environment and may have used less favourable sections for livestock farming. In Kosingen, subsistence was based primarily on pastoral agriculture supplemented by horticulture and reflects a close adjustment to the ecological potential. Cluster-analytical treatment of trace element data led to the formation of groups differing in diet, and we investigated whether this difference corresponded with social affiliation. In Neresheim, a non-random congruence of higher social status and access to high quality food was revealed, while such patterns were not observable in Kosingen. These interpopulational differences can be explained by bio-cultural interrelations between ecological properties of the natural units and reconstructed living conditions. PMID- 10816796 TI - Non-pharmacological treatments for atrial fibrillation. A critical perspective on the status quo. AB - In a proportion of patients with atrial fibrillation, antiarrhythmic drugs are either ineffective, unsafe, or poorly tolerated. Accordingly, a variety of non pharmacological treatments have been developed. This article critically reviews these modalities. (i) For ventricular rate control, catheter ablation of the atrioventricular node with pacemaker implantation is commonplace. An alternative is atrioventricular node modulation using a procedure similar to "slow pathway" ablation. (ii) For restoration of sinus rhythm, internal cardioversion using low energy shocks is highly effective; this has prompted the development of atrial and dual chamber defibrillators. (iii) To eliminate the atrial fibrillation substrate, a number of surgical procedures have been developed, of which the most effective is the "Maze" operation. The efficacy of this operation cannot be reproduced by conventional catheter ablation, and current research is concentrating on simplified procedures using new catheter designs for linear ablation. (iv) Finally, pacemakers and catheter ablation may be used to suppress the triggers for atrial fibrillation episodes. A number of atrial algorithms are under investigation for overdrive suppression of ectopy, and the use of multisite atrial pacing to alter the atrial response to ectopy has shown promising results. Catheter ablation has shown considerable success in preventing "focal" atrial fibrillation that is triggered or driven by ectopy arising usually from the pulmonary veins. To date, there are few data regarding the long-term efficacy and safety of these techniques, and their effects on quality of life. However, ongoing multicentre trials addressing these issues are expected to report over the next few years. PMID- 10816797 TI - [Present concepts of congenital long QT syndrome]. AB - The congenital long QT syndrome is characterised by the presence of syncopes due to torsades de pointe which may degenerate to ventricular fibrillation and cause sudden death. These syncopes occur in young subjects with electrocardiographic abnormalities and prolongation of the QT interval. Patients with the autosomally dominant transmitted Romano-Ward syndrome with normal audition are classically opposed to those with the Jervell and Lange-Nielsen autosomally recessive syndrome who have bilateral total deafness. Our understanding of the congenital long QT syndrome has improved in recent years with respect to the physiopathology, diagnosis and treatment, due to research in the fields of genetics, electrocardiography and electrophysiology. The diagnosis is based on analysis of the phenotype and genotypes. A family enquiry is always necessary to detect unrecognised forms. Five culprit genes have been identified for the Romano Ward syndrome. All code for subunits of sodium or potassium channels: two a subunits of the potassium channels (QVLQT1 for LQT1, HERG for LQT2), the a subunit of the sodium channel INa (SCN5A for LQT3), and two regulatory subunits of potassium channels (KCNE1 for LQT5 regulating the KvLQT1 channel and MiRP1 regulating HERG). The concept of genetic heterogeneity of the congenital long QT syndrome may thus be understood: different genes may be responsible for the same phenotype. Except for specific cases, the usual treatment is life-long betablocker therapy and the avoidance of a large number of drugs, the list of which is continually updated. A multicentre trial is underway to validate betablocker therapy for the prevention of cardiac events in a LQT1 genotype population. Prospective studies will be necessary to assess gene-specific treatments. PMID- 10816798 TI - [New invasive cartography techniques in electrophysiology]. AB - Fulguration, a widely used technique in the 80s, has been replaced by radiofrequency ablation. The limited and better controlled intra-cardiac lesions obtained by this method allowed the elimination of a large number of tachycardias with a high success rate. However, there are many arrhythmias, particularly at the atrial level, which are so complex that a limited analysis at a number of points of endocardial recording is inadequate. Recently, a number of systems has been introduced which considerably increased the acuity of observation of arrhythmias and the efficacy of their ablation. First of all, there are improved techniques of classical electrophysiological recording. Then, new systems of temporo-spatial recordings were developed and are described in this article. These use computer systems and enable particularly reliable and detailed approaches to cardiac anatomy and electrophysiology. However, the optimal use of these techniques can only be in specialised centres, by highly trained cardiologists in classical electrophysiology, for which they are complementary to rather than substitute for. PMID- 10816799 TI - [Left bundle branch block. Electrocardiographic and prognostic aspects]. AB - The concept of left bundle branch block (LBBB) was recognised at the beginning of the 20th century but confusion, due to the extrapolation data from animal experimentation, persisted for many years between the electrocardiographic appearances of LBBB and right bundle branch block (RBBB). The typical appearances of LBBB are now well known and consist of: 1) increased duration of the QRS complex > 0.12 seconds; 2) a wide, exclusive R wave with a plateau or notched summit in the left precordial leads and usually in D1 and aVL; 3) an important delay in the intrinsecoid deflection in the left precordial leads (0.08 to 0.12 seconds after the onset of QRS); 4) an axis of repolarisation opposite that of the QRS complex with so-called "secondary" abnormalities. The authors emphasise that some electrocardiographic variants carry a poor prognosis, in particular those with major QRS axis deviation to the left or, much less commonly, to the right. The diagnosis of left ventricular hypertrophy is possible in cases of LBBB by using the criteria of QRS amplitude in the left precordial leads. On the other hand, the diagnosis of myocardial infarction is more difficult, the criteria being very specific but having a sensitivity < 50%. The deleterious effects of LBBB on the haemodynamics are well known but their study has become a new firld of research since the introduction of bi-ventricular pacing for the treatment of cardiac failure. In dilated cardiomyopathy, LBBB increases the duration of functional mitral regurgitation and decreases left ventricular filling times. The prognostic implications of LBBB have been the object of many studies: the reports in the literature indicate a large increase in mortality when LBBB develops in patients over 44 years of age. The progression to complete atrioventricular block is common only when the HV interval exceeds 100 ms. In other cases, the prophylactic implantation of a cardiac pacemaker does not improve the prognosis which depends on the severity of the underlying cardiac disease. PMID- 10816800 TI - [Electrophysiologic and anatomic atrial remodeling: an atrial rhythmic cardiomyopathy]. AB - In 1995, Wijffels and Alessie, using a curious goat model of atrial fibrillation, introduced the concept of atrial remodelling. The classical atrial substrate (anatomopathological-dilatation and hypokinesis-, and electrophysiological-short refractory periods and decreased conduction-), appeared not only to be one of the causes of atrial fibrillation but also the consequence of atrial fibrillation itself, the mechanism being a vicious circle. In addition to ventricular rhythmic cardiomyopathy, responsible for cardiac failure, the concept of atrial rhythmic cardiomyopathy with the same mechanical and electrophysiological consequences, has developed. These changes, characterised mainly by calcium overload associated with cellular hibernation and differentiation, have not been totally elucidated but have already renewed the physiopathology of atrial fibrillation. PMID- 10816801 TI - [Syncope, conduction disorders. Controversial indications for cardiac stimulation]. AB - The objective of permanent pacemaker implantation is to provide against an increased risk of death or to improve quality of life by abolishing symptoms. In both cases, certain indications for pacing have clearly demonstrated to be strongly beneficial in well selected patients, but other are still controversial, due to the lack of convincing and converging published data, or to the absence of general consensus among specialists, or because selection criteria for pacing have been poorly defined. We try to clarify when to pace or not to pace in such conditions as first degree AV block, type I second degree AV block, intracardiac conduction defects, including those occurring at the acute stage of myocardial infarct or after cardiac surgery, sick sinus syndrome in cardiac transplant recipients, carotid sinus syndrome, vasovagal syncope, and unexplained syncopes. PMID- 10816802 TI - [Brugada's syndrome]. AB - Sudden death by ventricular fibrillation is a possible complication of most cardiac diseases but there are some cases of ventricular fibrillation assumed to be idiopathic, in which the most complete aetiological investigations remain negative. Syncope and sudden death by torsades de pointe then ventricular fibrillation of long QT syndromes without cardiac disease are well known: they are purely electric abnormalities, the ionic and genetic bases of which are becoming progressively better known. In 1991, a new syndrome was described by Pedro and Joseph Brugada with particular electrocardiographic appearances (right bundle branch block with ST elevation in leads V1-V3) and episodes of ventricular fibrillation occurring in patients without apparent heart disease. This syndrome, clinically similar to the unexpected cases of sudden death during their sleep of young men from South East Asia or living there, has aroused much interest. Similarly to the congenital long QT syndrome, the concept of a purely arrhythmic abnormality seems apparent and our understanding of the sequence leading from the electrocardiographic changes to the underlying ionic abnormalities initiating them and the genetic disorders which program them, have made great strides. PMID- 10816803 TI - [Role of antiarrhythmics in the post-infarction period]. AB - In the thrombolytic era, the mortality of myocardial infarction has been considerably reduced. The prognosis has also improved due to early treatment and the correction of residual ischaemia. Betablockers are valuable antiarrhythmic agents, both in the acute and chronic phases of infarction. Irrespective of the size of the infarct scar, a better prognosis is observed in patients taking betablockers. Class I antiarrhythmics, though, should be proscribed after the results of the CAST studies: these antiarrhythmics are effective on ventricular arrhythmias but do not improve the prognosis because of their proarrhythmic effects aggravated by ischaemia or left ventricular dysfunction. Of the Class III antiarrhythmics, amiodarone has been shown to reduce the incidence of sudden death in the post-infarction period in patients with ventricular hyperexcitability or severe left ventricular dysfunction. At present, classical antiarrhythmic therapy is opposed to the implantation of an automatic defibrillator in cases of serious arrhythmias after myocardial infarction. PMID- 10816804 TI - [Coronary revascularization: the design and the duty ... republican]. PMID- 10816805 TI - [Hospital results of salvage angioplasty with coronary stenting after failure of thrombolysis in the acute phase of myocardial infarction]. AB - Between December 1991 and February 19999, 25 patients (56 +/- 10 years) underwent salvage angioplasty with coronary stenting after failure of thrombolysis (TIMI 2), rtPA (N = 18), n-PA (N = 4), K2-tu-PA (N = 2) and streptokinase (N = 1). All were treated by aspirin and 96% were given ticlopidine for one month. The culprit artery was the left anterior descending (48%), the left circumflex (8%) or the right coronary (44%). The average ejection fraction was 41%; 4 patients (16%) had cardiogenic shock. The stents were implanted for occlusive coronary dissection (36%), threatening dissection (40%), partial result of angioplasty (20%) or of first intention (4%). In all, 31 stents were implanted (1.2 +/- 0.57 stent/target lesion ratio with an average length of 20.9 +/- 10.2 mm). The stents were tabular in 51% of cases. The angiographic success rate (TIMI 3 and residual stenosis < 50%) was 96% with maximum inflation pressures of 13.7 +/- 2.5 atm and balloons with an average diameter of 3.3 +/- 0.5 mm. Intra-aortic balloon pumping was required in 7 patients (28%). The 30 day results included a mortality rate of 16% (4 patients), a recurrence of infarction in 4%; there were no repeat angioplasties, coronary bypass surgery or blood transfusions. The predictive factors of recurrent coronary events were: age over 60 (p = 0.04), multivessel coronary disease (p = 0.007), cardiogenic shock (p = 0.004) and left ventricular dysfunction (p = 0.015). The authors conclude that cases of failure of thrombolysis are at high risk and that salvage angioplasty with coronary stenting is associated with excellent angiographic results. Patients with cardiogenic shock, however, have a high mortality, irrespective of coronary patency and the use of intra-aortic balloon pumping. PMID- 10816806 TI - [Percutaneous coronary angioplasty via the radial artery. Feasibility and safety]. AB - The authors report the results of percutaneous coronary angioplasty using the radial approach with respect to feasibility and safety. Between February 1994 and December 1998, out of a total of 9,070 coronary angioplasties performed in our Institute, the radial arterial approach was chosen in 5,354 cases (59%). During this period, the proportion of procedures performed via the radial artery progressively increased to 78% of cases in 1998. Cannulation of the radial artery was successful in 5,244 cases (98%). Rotatory atherectomy was used in 96 cases (2%), the excimer laser in 38 cases (1%) and stenting was performed in 3,350 cases (64%). Angiographic success was obtained in 5,224 patients (99%). Serious cardiac complications occurred in 82 patients (1.5%), including 17 deaths (0.3%), 22 Q wave myocardial infarctions (0.4%), 28 non-Q wave infarcts (0.5%), 5 coronary bypass procedures (0.1%) and 10 sub-acute occlusions (0.2%). The vascular complications included 1 death due to an ischaemic cerebrovascular accident, 25 haematomas at the puncture site (0.5%) and 3 surgical arterial repairs. The radial pulse disappeared after the procedure in 4% of cases, all asymptomatic. Angioplasty by the radial artery in the hands of trained operators is feasible and associated with a relatively low rate of cardiac and vascular complications. PMID- 10816807 TI - [Hematoma of the aortic wall: from diagnosis to treatment]. AB - Haematoma of the thoracic aortic wall is a relatively new concept, the physiopathology of which remains controversial. It results from an haemorrhage of the aortic wall due to rupture of the vasa-vasorum without communication with the arterial lumen. This is a diagnosis of elimination of dissection of the aorta which has been made possible by modern techniques of imaging, such as transoesophageal echocardiography, helicoidal scanner and magnetic nuclear resonance imaging. The prognosis of haematoma of the aortic wall is not as bad as that of dissection of the aorta. Recent studies have shown that the condition may stabilise, regress or progress towards complications of two types: early, dissection or fissuration of the aorta, and late, aortic aneurysm. This is a medico-surgical emergency, the treatment of which is not well codified. However, schematically, haematoma of the aortic wall should be managed in the same way as dissection of the aorta: surgery when the ascending aorta is affected, medical treatment in other cases in the absence of complications. PMID- 10816808 TI - [Cardiac beriberi]. AB - Interest has recently risen regarding thiamine deficiency (beriberi). In industrial countries, not only alcoholics, but also deprived people with malnutrition, elderly patients and patients with AIDS are at risk of thiamine deficiency. Moreover, long-term furosemide use may be associated with thiamine deficiency through urinary loss, contributing to cardiac insufficiency in patients with congestive heart failure. Cardiovascular (wet beriberi) manifestations of thiamine deficiency are characterized by peripheral vasodilatation with increased cardiac output, myocardial lesion, sodium and water retention and biventricular myocardial failure. Treatment consists of thiamine administration with rapid clinical improvement after supplementation. PMID- 10816809 TI - [Malignant mediastinal schwannoma in Recklinghausen's disease presenting as cardiac tamponade. Report of two cases]. AB - The authors report two cases of Recklinghausen's disease admitted for cardiac tamponade. Two-dimensional echocardiography showed a pericardial tumour in addition to a compressive pericardial effusion. After pericardial drainage, the outcome was rapidly fatal. Histological examination confirmed the diagnosis of a malignant intrapericardial schwannoma. A review of the literature revealed the scarcity of mediastinal malignant schwannoma. Associated with Von Reckinghausen's disease in 2 to 13% of cases, the prognosis is poor. The presentation of malignant mediastinal schwannoma with cardiac tamponade remains very rare. PMID- 10816810 TI - [Diagnosis of cardiac tumor by percutaneous thromboaspiration. A case report]. AB - Percutaneous endovascular thromboaspiration is a valuable tool as illustrated by the case of a patient suffering from a large intracardiac tumour. Histological and immuno-histochemical analysis of the tumour fragments provided the diagnosis of a cardiac angiosarcoma. The patient was a 44 year old man admitted for a large sero-sanguinous pericardial effusion which recurred after drainage. The case was complicated by a haemorrhagic cerebrovascular accident unrelated to a secondary deposit. Initially suspected after transthoracic echocardiography, the diagnosis of a tumour invading the right atrium was confirmed by transoesophageal echocardiography and cardiac CT scan. Surgery was declined and as the diagnosis of lymphoma could not be excluded, the patient underwent biopsy by an original method of percutaneous thromboaspiration. This minimally invasive, low cost technique would appear to be a valuable alternative to other endovascular biopsy techniques (saber, biotome) and to surgical biopsy, and could be proposed as the technique of first intent in an a priori non-operable intra-cardiac tumour or when lymphoma is suspected. PMID- 10816811 TI - [Coronary angioplasty, "mini-hospitalization" and cigarettes. Too often we can't see the coronary disease forest for the stenosis trees]. PMID- 10816812 TI - [Cardiovascular research in France]. PMID- 10816813 TI - [Modeling of peptides and proteins in a membrane environment.II. Structural and energetic aspects of Glycophorin A in a lipid bilayer]. AB - The conformational space of a hydrophobic peptide fragment of glycophorin A in a lipid membrane was studied with the Monte Carlo method using the solvation model described in the first communication of this series. The simulation was performed for various starting orientations of the peptide relative to the membrane bilayer: outside, inside, partially immersed, and transbilayer. We showed that the membrane substantially stabilizes the alpha-helical conformation of the central hydrophobic part of the glycophorin A molecule, which for the most part is immersed in the apolar core of the bilayer. For various conformational states, energy values were calculated and the orientations of the peptide relative to the membrane were characterized. Depending on the thickness of the bilayer, either an entirely alpha-helical conformation in transbilayer orientation or a conformation with a kink in the central part of the helix with the N- and C-termini exposed on one side of the membrane corresponds to the minimal-energy structure. The transmembrane orientation of glycophorin A is energetically advantageous when the membrane thickness is close to the length of its hydrophobic helical portion, which is consistent with the effect of "hydrophobic match" observed experimentally. The prospects for further refinement of the model are discussed. PMID- 10816814 TI - [Single amino acid substitutions in the Ca2+-binding site of recoverin.II.The unusual behavior of the protein upon the binding of calcium ions]. AB - The structural properties of myristoylated forms of recombinant recoverin of the wild type and of its mutants with damaged second and/or third Ca(2+)-binding sites were studied by fluorimetry and circular dichroism. The interaction of wild type recoverin with calcium ions was shown to induce unusual structural rearrangements in its molecule. In particular, protein binding with Ca2+ ions results in an increase in the mobility of the environment of Trp residues, in higher hydrophobicity, and in elevated thermal stability (its thermal transition shifts by 15 degrees C to higher temperatures) but has almost no effect on its secondary structure. Similar structural changes induced by Ca2+ are also characteristic of the -EF2 mutant of recoverin whose second Ca(2+)-binding site is modified and cannot bind calcium ions. The structural properties of the -EF3 and -EF2,3 mutants (whose third or simultaneously second and third Ca(2+)-binding sites, respectively, are modified and damaged) are practically indifferent to calcium ions. PMID- 10816815 TI - [Mapping of active site of alcohol dehydrogenase with low-molecular ligands]. AB - In search of an active alcohol dehydrogenase inhibitor, the structure of which may serve as the basis for a potential drug design, the active site of alcohol dehydrogenase containing NAD and Zn2+ ions was mapped using the method of molecular mechanics. Molecular docking was performed using a number of ligands containing characteristic functional groups: formate ion, ammonia, ammonium ion, methanol, and methylamine. Sites of preferable binding were revealed for each ligand and arranged in order of decreasing energy of binding to the enzyme. A comparison of the predicted ligand-binding sites and the experimental data on the location of water and inhibitor binding sites in the known structures of corresponding alcohol dehydrogenase complexes indicated a coincidence of the complex formation sites, which confirms the validity of the method and provides the requirements for a highly effective inhibitor (the pharmacophore model). PMID- 10816816 TI - [Identification of the protease responsible for the antiadhesive properties of dog blood serum]. AB - The antiadhesive effect of dog blood serum described previously was shown to be associated with the proteolytic activity of the serum components. The protease exhibiting this antiadhesive effect was isolated by several fractionation stages and identified with plasmin. PMID- 10816817 TI - [A study of aspartyl proteases using intramolecularly quenched fluorogenic peptide substrates]. AB - A series of fluorogenic tetra-, penta-, and hexapeptide substrates of the general structure Abz-X-Phe-Phe-Y-Ded (or -pNa in place of -Ded), where X = Ala, Ala-Ala, or Val-Ala and Y = -, Ala, or Ala-Ala, were proposed. Kinetic parameters of hydrolysis of these substrates by pepsin, cathepsin D, human gastricsin, pig pepsin, calf chymosin, and aspergillopepsin A were determined. The compounds synthesized proved to be effective substrates for aspartyl proteases of diverse origins. PMID- 10816818 TI - [Synthesis and properties of photolabile caged phosphotriester derivatives of dinucleoside phosphates]. AB - Dinucleoside phosphates that harbor phosphate groups transiently blocked (caged) by o-nitrobenzyl or o-nitroveratryl residues were synthesized. It was shown that the conditions of the UV-induced deprotection largely depend on the nature of the protective group. The phosphotriesters obtained were resistant toward snake venom phosphodiesterase and nucleases of the cellular extract. The synthesis of the dinucleoside phosphates containing a photolabile group preceded the incorporation of the modified blocks into extended oligonucleotides by the phosphoramidite method. PMID- 10816819 TI - [Synthesis of disaccharide Neu5Gcalpha(2-6)GalNAcalpha as a spacer glycoside]. AB - The first synthesis of the Neu5Gc analogue of SiaTn disaccharide, which can be detected in breast tumors by immunochemical methods, is reported. The regioselective sialylation of (3-trifluoroacetamidopropyl)-2-azido-2-deoxy-alpha D-galactopyranoside with peracetate of the methyl ester of N acetoxyacetylneuraminic acid beta-ethylthioglycoside in the presence of N iodosuccinimide and trifluoromethanesulfonic acid (or its trimethylsilyl ester) resulted in the derivatives of alpha- and beta-sialyl(2-->6)galactosaminide in 39 and 32% yields, respectively. The catalytic hydrogenolysis of the azido group and subsquent N- and O-acetylation of the alpha-anomer gave the trifluoroacetamidopropyl glycoside peracetate. Removal of the protective groups led to glycoside Neu5Gc alpha(2-->6)GalNAc alpha-O(CH2)3NH2. Using the Neu5Gc derivative with acetoxyacetyl groups at positions O9 and O4 as a donor increases the alpha-selectivity of sialylation to afford the alpha- and beta-anomers in 69 and 8% yields, respectively. PMID- 10816821 TI - [The study of hydrolysis of new lipid-like substrates and trilaurin in monolayers catalyzed with the lipase from Pseudomonas fluorescens]. AB - A simple method for determining the enzymic hydrolysis parameters of lipid-like substrates and trilaurin assembled in monolayers at the water-air interface was suggested. At a surface pressure of 10 mN/m, the initial rates of lipolysis were found to be proportional to the decrease in area of the substrate monolayer caused by the enzymic hydrolysis in a single-compartment Langmuir balance. The kinetic parameters for the hydrolysis of trilaurin and three 1,3 dilaurylpseudoglycerides acetylated in position 2 with an amino acid (phenylalanine, leucine, or valine) catalyzed with lipase from Pseudomonas fluorescens were determined. Unlike models of enzymic hydrolysis that neglect the thickness of the substrate monolayer, our method allows the determination of kinetic parameters in standard dimensions. The values of kcat for the synthetic pseudoglycerides were found to be significantly higher than that for trilaurin, while the values of Km(app) were close. This may be due to the presence of positively charged primary amino groups in the molecules of pseudoglycerides. PMID- 10816820 TI - [Synthesis of the lupane group triterpenoids and there hepatoprotective activity]. AB - Hemisuccinates, hemiphthalates, acetylsalicylates, cinnamates, and p methoxycinnamates of lupeol, betulin, and 3-O-acetylbetulin were synthesized via interaction with corresponding acid anhydrides or acid chlorides. A number of betulin esters in position 3 and 28 were shown to exhibit a pronounced hepatoprotective effect similar to that of betulin and silibor. These experimental data were in a good agreement with the computer prediction of their biological activity. Betulin 3,28-bis-hemiphthalate was more effective than carsil in models of experimental hepatitis caused by carbon tetrachloride, tetracycline, and ethanol. PMID- 10816822 TI - [A noninstrumental Immunoassay based on colloidal dyes]. AB - Detecting labels based on water dispersions of colloidal textile dyes were developed that are useful in various analytical and diagnostic test systems for a simple visual assessment of the assay. Colored water-insoluble particles of dyes were used for the sorptional immobilization of streptavidin on their surface. The resulting streptavidin-dye (STR-DYE) complexes possessed a high visualizing capacity and were used for the combined detection of pesticides (simazine and 2,4 dichlorophenoxyacetic acid) by noninstrumental immunoassay (DYE-comb-assay, competitive dot-immunoassay in the comb format). The detection limits and the duration of our DYE-comb-assay (4 ng/ml, 20-25 min), HRP-comb-assay (competitive dot-immunoassay in the comb format using the enzymic conjugate of STR with horseradish peroxidase) (16 ng/ml), and the traditional competitive ELISA (12-16 ng/ml, 1.5 h) were compared. This DYE-comb-assay is simple enough and can be used under field conditions. PMID- 10816823 TI - [Degradation of phosphatidylnucleosides induced by phospholipases]. AB - Hydrolysis of phosphatidylnucleosides, 5'-(rac-1-hexadecyl-2-palmitoyl-sn-glycero 3-phosphoryl)-3'-azido- 3'-deoxythymidine and -2',3'-didehydro-3'-deoxythymidine, effected by phospholipases (PL) A2, C, and D was studied to reveal the metabolism of these derivatives. It was shown that PLA2 deacetylates the glycerol residue at position 2, PLC is inactive, and PLD hydrolyzes the phosphatidylnucleosides to give free nucleosides. PMID- 10816824 TI - Vaccine manufacturer should not be implicated. PMID- 10816825 TI - Comments on practising complementary and alternative modalities. PMID- 10816826 TI - Simple facts and numbers. PMID- 10816827 TI - Need to distinguish between studies designed to give an unbiased answer and those not so designed. PMID- 10816828 TI - An ethicist's commentary on whether ineffective medication should be permitted to be sold. PMID- 10816829 TI - Professionalism--a code of behavior. PMID- 10816830 TI - Equine phacoclastic uveitis: the clinical manifestations, light microscopic findings, and therapy of 7 cases. AB - This retrospective clinical study describes the clinical manifestations, light microscopic findings, and diagnosis and treatment of acute and chronic lens rupture in the horse. Rupture of the lens capsule in the horse usually results in a chronic, blinding inflammation (phacoclastic uveitis) unless prompt surgical and medical therapies are implemented. The clinical manifestations of acute lens capsule rupture included: cataract; intralenticular displacement of iridal pigment; lens cortical fragments attached to the perforated lens capsule, iris, and corneal endothelium; miosis; aqueous flare; and usually a corneal or scleral perforation with ulceration or focal full thickness corneal edema and scarring. The clinical signs of chronic phacoclastic uveitis include blindness, phthisis bulbi, and generalized corneal opacification related to scarring, vascularization, pigmentation, and edema. In one horse, acute phacoclastic uveitis was successfully treated with phacoemulsification to remove the ruptured lens and medical therapy to control the accompanying inflammation. The affected eyes of the horses with chronic phacoclastic uveitis were enucleated because of persistent clinical signs of nonulcerative keratitis and uveitis, despite long term medical management. The clinical manifestations and lack of improvement with medical therapy are similar in the horse, dog, cat, and rabbit. However, the histologic findings in equine phacoclastic uveitis differ significantly from those in the dog, and rabbit. PMID- 10816831 TI - Epidemiological study of Yersinia enterocolitica in swine herds in Quebec. AB - The objectives of this study were the identification of the different contamination sources of Yersinia enterocolitica, as well as the determination of the prevalence and the distribution of the different genotypes in swine herds. The owners of 20 farms, located in the Richelieu-Yamaska region, agreed to participate in the study. Each farm was visited a minimum of 5 times between May and October 1997, and, at each visit, 20 environmental and 10 fecal samples were collected. Yersinia enterocolitica isolates were identified, serotyped, and submitted to a genetic characterization by pulsed-field gel electrophoresis. The correlation coefficient (0.61) between prevalence in environment and in feces was significant (P = 0.004). Among the 153 positive samples, 93.5% belonged to serotype 0:3. The comparison of PFGE profiles revealed that all environmental Y. enterocolitica isolates had a profile identical to that of isolates recovered in feces from the corresponding farms. Also, when the genetic profiles of isolates recovered from feces collected at the first visit were compared with the profiles of isolates obtained from the subsequent visits, the same profile was observed on every farm. We concluded that environment does not represent the main source of contamination of swine by Y. enterocolitica and that, in most instances, the same strain persists in a barn from one production lot to another. PMID- 10816832 TI - Use of ionophores in lactating dairy cattle: a review. AB - Ionophores are feed additives that alter rumen microbial populations through ion transfer across cell membranes. Although ionophores have been used widely in the beef industry for improved feed efficiency and control of coccidiosis, there has been limited use by the dairy industry. In Canada, the label warning prohibiting the use of monensin premix in lactating dairy cattle was removed in June 1996. Following this, in December 1997, a controlled release capsule containing monensin was approved for use in dairy cattle as an aid to prevent subclinical ketosis. Monensin may have several advantages for dairy cattle, including improved energy metabolism, increased milk production, and altered milk components. This literature review was primarily conducted in 1996 by using the Agricola and CAB search databases. Other relevant articles published since the search (up to 1998) have been added. This review will provide practitioners with relevant references in the published literature regarding ionophore use in dairy cattle. It should also give some guidance as to what effects might be anticipated with the use of ionophores in lactating dairy animals. PMID- 10816833 TI - Ligature slippage during standing laparoscopic ovariectomy in a mare. AB - Suture ligature failure is a potential complication during laparoscopic ovariectomy techniques utilizing ligatures as a means of hemostasis. This complication in the standing mare and the successful use of laparoscopic electrosurgical instrumentation as the sole means of providing hemostasis to the mesovarium of a mare are described. PMID- 10816834 TI - Production of sexed lambs after biopsy of ovine blastocyts produced in vitro. AB - Embryos were generated by in vitro fertilization of in vitro-matured oocytes, cultured to the blastocyst stage, biopsied for sex determination by a PCR-based procedure, and transferred to synchronized recipients. Three out of 5 sheep (60%) were diagnosed pregnant, and 4 lambs of predicted sex were born. PMID- 10816835 TI - Eccrine adenocarcinoma of the footpads in 2 cats. AB - Adenocarcinoma of sweat glands of the footpads was diagnosed in 2 cats. Clinical signs included lameness and swelling of multiple digits. Pulmonary metastasis was detected in one case. Diagnosis was based on histopathological and immunohistochemical findings. Eccrine adenocarcinoma should be included in the differential diagnosis of footpads lesions in aged cats. PMID- 10816836 TI - Velogenic Newcastle disease in imported caged birds. AB - Velogenic Newcastle disease was diagnosed in pet birds intended for importation into Canada. Virological and histopathological examination confirmed the presence of the disease. The group of birds was denied entry into Canada. Similar birds illegally imported are a potential source of velogenic Newcastle disease virus and are a threat to domestic poultry. PMID- 10816837 TI - Recurrent intussusception in a 14-month-old, spayed, female German shepherd cross. PMID- 10816838 TI - Tansy ragwort poisoning in a horse in southern Ontario. AB - Bizarre behavior, apparent lameness, and colic were noticed in 1 of 3 horses on a pasture overgrown by weeds during a drought. Liver failure and hepatoencephalopathy were diagnosed, caused by pyrrolizidine alkaloid toxicosis associated with consumption of tansy ragwort. The horse made a full recovery when removed from the pasture. PMID- 10816839 TI - Salmonella typhimurium DT 104 in sheep. PMID- 10816840 TI - Distribution of Streptococcus suis capsular types in 1999. PMID- 10816841 TI - The unconscious and psychosis. Some considerations on the psychoanalytic theory of psychosis. AB - The author contends that the various psychoanalytic theories and techniques employ different models of the unconscious, each relating to a different unconscious reality describable in terms of specific mental functions. He reviews in particular the Freudian dynamic unconscious, based on repression; the Kleinian unconscious, which adds the notions of unconscious fantasy and splitting of the object; Bion's conception of the unconscious as a mental function of which the subject is unaware but which can formulate thoughts and metabolise emotions; and the neuroscientific view of the unconscious as coinciding with that of which one is unaware and not with the Freudian repressed. The author thus distinguishes between the dynamic and the emotional unconscious and between 'unconscious' and 'unaware', and notes the role of distortion of the 'unaware' perceptions involved in the analytic relationship in the impasse situation. He is particularly concerned to show that, whereas neurosis involves the dynamic unconscious, psychosis alters the emotional unconscious, the entity underlying the sense of identity and the 'unaware' consciousness of existence. In psychosis the emotional unconscious is blinded, so that the patient is conscious but lacks awareness. The dynamic unconscious is also affected. After presenting two case histories, the author draws attention to the need for further clinical and theoretical research in this field. PMID- 10816842 TI - The psychoanalyst and the transsexual patient. AB - The author remarks on the dearth of psychoanalytic literature concerning analyses of transsexuals and of clinical experience with such patients in general. Drawing on her personal work with transsexual patients at a specialised centre, as well as with children suffering from gender identity disorders and their parents, she is able to specify the factors which, in her view, make the psychoanalysis and psychotherapy of these subjects so difficult. In particular, they are totally focused on the body and on their intention of securing sex reassignment by hormonal and surgical treatments, so that they rule out the involvement of any psychic element. The psychic functioning of these patients is illustrated by some clinical vignettes, and the transference and countertransference problems are discussed. The author shows how the patients concerned have great difficulty in accepting a psychological approach to their problems; they do not speak the language of wishes and conflict, and claim to remember nothing of their childhood or past life. She concludes that transsexualism is a narcissistic disorder in which the constitution of the self has been profoundly impaired and that it is only since analysis have embarked on the treatment of non-neurotic patients that the condition has become accessible to psychoanalytic psychotherapy or indeed to psychoanalysis. The difficulties notwithstanding, the author considers that such work is worthwhile and that patients can benefit from it. PMID- 10816843 TI - Sexuality and perversion a hundred years on: discovering what Freud discovered. AB - The author considers how Freud's original formulation of the sexual drive may be understood in contemporary terms. Parallels between psychoanalysis, art history and the social and natural sciences underline the idea of sexuality as a construct that can be revisited. The nature of sexuality depends on how its irreducible biological basis is interpreted by a particular society at a particular time. The importance of revisiting is illustrated by Klein's concept of a position, which revisits the theoretical construct of a developmental stage. Freud's own construction of sexuality is based on his view of drive as comprising source, aim and object. Freud, in the Darwinian climate of his time, considered the object in terms of whether it served the fulfillment of the aim, but his paradigm can today be understood more broadly than Freud himself was able to. A discussion of perversion shows that psychoanalysis has moved from seeing it as defensive against instinctual derivatives to seeing it as defensive against object-relatedness. This reflects a shift in the analytic conceptualization of sexuality in general. Freud's paradigm for the sexual drive can be reconceived in terms of source, aim and quality of relatedness to object. Such a view of sexuality also has implications for gender issues, for the comparison of male and female perversion, and for the relation between sexuality and love. PMID- 10816844 TI - Some views on the manifestation of the death instinct in clinical work. AB - The author argues that the postulation of the instinctual drive towards death can be seen as an attempt to account for the manifestation of a destructive psychological force that is palpably present in many of our patients. He discusses some of the experiences, activities and aims that reflect this destructive psychic force, and the conscious and unconscious gratification that is intrinsically bound up with it. What is 'deadly' is the way in which meaning, specificity and differences are attacked, and any developmental processes retarded or undermined. The vitality is taken out of the patient himself as well as his objects, and although in an important sense these drives are 'anti-life', the author suggests that their aim is not literally to kill or to annihilate, but that the patient feels compelled to maintain a link with the object that often has an evidently tormenting quality. Using a detailed clinical illustration, the author argues that the gratification that is bound up in these activities, and which gives them such a compulsive quality, does not result from fusion with the life instinct, with the resultant libidinisation of the death instinct. On the contrary, the gratification obtained from attacking, spoiling and undermining, whether directed to the self or the object, is an essential element of such a destructive drive. PMID- 10816845 TI - Interpretation and containment. AB - The author explores two aspects of the analyst's effort to imagine the inner world of his patient and the way that they are manifest in the clinical moment. The first of these is the analyst's recognition and interpretation of his patient's elaborated fantasies. This current of the analyst's imagination is most often evoked by the patient's communication of whole-object transferences, which occurs largely in his verbal associations. The second is the analyst's reception and transformation of his patient's primitive emotional experience, a process that Bion has called containment. This second imaginative current is most often evoked by the patient's communication of part-object transferences, which occurs largely in affect and action. Interpretation and containment both go on at once in clinical work, although one or the other is usually dominant. Attention to the interplay of interpretation and containment in the clinical moment enables us to identify the articulation of whole- and part-object transferences and to integrate ego-psychological and Kleinian frames of reference in clinical work. In addition, the concept of mutual containment opens Kleinian theory to the possibility of a two-person psychology in which the roles of analyst and patient are more symmetrical than they are usually conceived to be within this frame of reference. The author presents two clinical examples to demonstrate the interplay of interpretation and containment. In the first, these processes operate smoothly. In the second, the process of containment is strained but ultimately successful. PMID- 10816846 TI - Reflections on 'thinking in the presence of the other'. AB - The author argues that as analysis progresses, mutual processes of projective and introjective identification result in an interpretation of analyst and patient such that the contributions that each participant makes to the analytic dialogue are to a significant degree imbued with the prior contributions of the other. This consideration renders the concepts of evidence and confirmation ambiguous and applicable only with a certain amount of caution. Further complexity is added to analytic protocols by the limited regulation of interpretation by the general guidelines of the theoretical orientation in play. This understanding of the analytic process contrasts sharply with that put forward by those who place particular emphasis on the centrality of the so-called real relationship in bringing about change. PMID- 10816847 TI - A concerned critique of psychoanalytic education. AB - The author explores some central problems in contemporary psychoanalytic education. He compares strengths and limitations of the two dominant models of psychoanalytic training, the traditional Eitingon model and the French model, and reviews current efforts to modify these models in order to overcome their limitations. In examining problems common to both educational systems, the author highlights the following issues: a tendency to infantilise psychoanalytic candidates, a persisting trend towards isolation from the scientific community, a lack of consistent concern for the total educational experience of candidates, authoritarian management and a denial of the effects of external, social reality on psychoanalytic education. Proposed solutions to these problems include: a stress on 'step-by-step' evaluation of candidates' progression, a greater emphasis on the cognitive aspects of seminars and supervision, particularly, a systematic exploration of the psychoanalytic method and its applications, a re examination of the usefulness of the function of the training analyst status, an integration of teaching and practical experience in systematic research, and the incorporation of contemporary educational methods as part of the strengthening of the academic ambience of psychoanalytic institutes. The author concludes with a list of fifteen questions that may provide a quick indication of how for a psychoanalytic institute has progressed with the work of educational innovation. PMID- 10816848 TI - The primal scene and Picasso's Guernica. AB - The author examines a group of works by Picasso dating from the late 1930s in terms of the artist's experiences as documented by his biographers and of primal scene fantasies as described in the field of psychoanalysis by, in particular, Freud and Klein. Pointing out that the artist himself is on record as inviting such a consideration, he contends that these fantasies constitute the latent motivating force behind one of Picasso's most famous paintings, the mural Guernica, and a number of other productions from the same period. Biographical accounts are drawn upon to show how aspects of his inner world are revealed in the specific works described and reproduced in this paper. The role of women is shown to have been particularly relevant. The author demonstrates how Picasso's constant pattern of triangular relationships culminated in his personal crisis of 1935, which, together with the Spanish Civil War, reflecting as it did the conflicts of his internal and external relations, contributed to the production of the works in this group. The artist is seen as attempting to work through and make reparation for envious attacks on the parental objects, but it is pointed out that art works should not be assessed by the criterion of therapeutic change. PMID- 10816849 TI - Biological and integrative studies on affect. PMID- 10816850 TI - Holocaust: affect and memory. PMID- 10816851 TI - Affect and psychosis. PMID- 10816852 TI - Development of affect in bilingual patients. PMID- 10816853 TI - Pleasure in phantasy and reality. PMID- 10816854 TI - Affect, somatization and symbolization. PMID- 10816855 TI - Art, affect and aesthetics. PMID- 10816856 TI - Affective self-disclosure by the analyst. PMID- 10816858 TI - Incommensurable worlds. PMID- 10816857 TI - The recovered memories controversy. PMID- 10816859 TI - 'Dreams that turn over a page...'. PMID- 10816860 TI - 'Relating to the superego' by Edna O'Shaughnessy. PMID- 10816861 TI - Two cultures in medicine. PMID- 10816862 TI - Withdrawing and withholding life-prolonging medical treatment from adult patients. PMID- 10816863 TI - How can we afford costly medicines? AB - Affording the ever-increasing cost of medicines is a struggle throughout the NHS. A College Working Party has addressed the issues in an extensive recent report that analyses the cost of medicines, their licensing and evaluation, priority setting and its ethical and legal implications. It considers options for increasing the available resource, but explains why everyone concerned with the evaluation, funding and use of medicines must recognise the need to set priorities. Though the report does not pretend to have solved the problem facing the NHS, it describes many examples of good practice and suggests a way forward. PMID- 10816864 TI - The pharmaceutical industry and the clinical professions. PMID- 10816865 TI - The National Clinical Guidelines for Stroke. PMID- 10816866 TI - The adolescent with respiratory disease. PMID- 10816867 TI - Nutrition, eating and gastrointestinal conditions in adolescence. PMID- 10816868 TI - Abnormalities of growth and development in puberty. PMID- 10816869 TI - Musculoskeletal diseases in adolescence. PMID- 10816870 TI - Congenital heart disease in adolescence. AB - Adolescents and young adults with congenital heart disease are increasingly commonly seen in cardiological practice. Their problems are particular and often unique. Their management requires training and experience in the management of congenital heart disease by the attending cardiologist. In these days of clinical governance, management of ACHD patients must be supervised by clinicians specifically trained in handling congenital cardiac malformation. PMID- 10816871 TI - Adolescent nephro-urology. PMID- 10816872 TI - Alcohol abuse and the burden on the NHS--time for action. AB - Alcohol misuse is a major drain on the resources of the NHS. Strategies that are adequately resourced are urgently needed to be able to cope with the burden of alcohol-related illness. PMID- 10816873 TI - Designing meningitis vaccines. AB - Conjugate polysaccharide vaccines are a recent intervention to combat the relative inability of young children to mount an effective immune response against encapsulated bacteria, especially Haemophilus influenzae (Hib), Neisseria meningitidis (Nm) and Streptococcus pneumoniae (Sp). These organisms cause the majority of community acquired septicaemia and meningitis in UK children. Their capsular polysaccharides, important virulence factors in evading phagocytosis, are poorly immunogenic in young children compared to adults. Conjugation, by covalent linking, of the polysaccharide to an immunogenic protein, has been demonstrated for each of these organisms to produce good antibody response to the polysaccharide. Conjugate Hib vaccines have proven effective in reducing Hib meningitis and invasive disease in the countries that have introduced them. Pneumococcal conjugate vaccines have proven effective in preventing invasive disease caused by serotypes contained in the vaccines. Efficacy studies are awaited for meningococcal conjugate vaccines. PMID- 10816874 TI - Behcet's syndrome. AB - We report our clinical experience of 230 patients referred to the Hammersmith hospital with a working diagnosis of Behcet's syndrome. The pathogenesis, diagnosis and management of the syndrome are discussed. PMID- 10816875 TI - Selection bias in the management of unstable angina. AB - OBJECTIVES: To examine the criteria for selecting patients presenting with unstable angina for cardiac catheterisation and to assess the extent to which these criteria successfully incorporate high risk groups. METHODS AND RESULTS: This was a prospective cohort study of 517 patients admitted with unstable angina with 12 months follow-up; 139 patients (26.9%) had cardiac catheterisation 32 days or longer after presentation. The odds of early catheterisation were increased by regional ST segment depression on the presenting ECG (odds ratio (OR) 1.70, 95% confidence intervals (CI) 1.01-2.87) and ongoing ischaemic chest pain more than 12 hours after admission (OR 9.72, CI 6.10-15.49), and reduced by age over 65 years (OR 0.56, 95% CI 0.35-0.90) and heart failure (OR 0.26, CI 0.11 0.64). The 12-month rates of myocardial infarction (MI) or death were 8.6% and 17.7% (p = 0.01) in patients who were and were not referred for early cardiac catheterisation, respectively. Survival analysis showed that the odds of MI and death in the first 12 months were increased substantially by heart failure (OR 2.82, 95% CI 1.53-5.20) and age over 65 (OR 1.91, 95% CI 1.13-3.23). CONCLUSION: Selection for early cardiac catheterisation in this unstable angina population was largely ischaemia-driven, based on ongoing chest pain and ST segment depression. This policy was associated with a low event rate in the ischaemic group, but it failed to target elderly patients and those with heart failure who were at greatest risk of MI and death during the first year. PMID- 10816876 TI - Charter for disabled people using hospitals: a completed access audit cycle. AB - This completed audit cycle assessed access for disabled people to a district general hospital, in relation to standards laid down in the Royal College of Physicians Charter for Disabled People using Hospitals. The project was effective in demonstrating problems and implementing change to overcome them. It was also useful in raising disability awareness in the young investigators, who easily recognised the shortcomings in facilities for disabled people, and is proposed as a possible model for inclusion in medical undergraduate training programmes to raise disability awareness amongst a new generation of doctors. PMID- 10816877 TI - Lectins, colitis and colon cancer. PMID- 10816878 TI - Colorectal cancer. PMID- 10816879 TI - Clinical research funding: a priority for the Medical Research Council. PMID- 10816880 TI - Inflammation in cardiovascular disease. PMID- 10816881 TI - Epics in intensive care: acute asthma. PMID- 10816882 TI - National sentinel audit on stroke--cognitive assessments. PMID- 10816883 TI - Congestive heart failure--can the nephrologist help? PMID- 10816884 TI - Adolescent medicine. PMID- 10816885 TI - What the SHO really does. PMID- 10816886 TI - SHOs' experience of practical procedures. PMID- 10816887 TI - Telephone access to healthcare. PMID- 10816889 TI - Views of junior medical staff who work full shift rotas. PMID- 10816888 TI - Botulinum toxin follow-up clinics: training implications. PMID- 10816890 TI - [Generations-conscious policy does not condone financial constraints to health services]. PMID- 10816891 TI - [Nursing home rehabilitation, functional independence and quality of life: a pilot study of somatic nursing home rehabilitation patients]. AB - In a pilot study of 18 patients the relation between nursing home rehabilitation, functional independence and quality of life was examined. Furthermore, measurement instruments and study design were evaluated. Nursing home rehabilitation proved to be associated with (partially significant) improvement in functional independence and quality of life. Provided the inclusion of a control group and an observation period of adequate duration, study design and measurement instruments seemed to be appropriate for application in a multicenter study. PMID- 10816892 TI - [Model of costs of care for dementia: community-dwelling vs. institutionalization]. AB - This article describes a model of costs for Alzheimer's Disease. The cost of institutionalization and the cost of living at home for patients with Alzheimer's Disease are calculated and compared. The average yearly cost of living at home is 10,810 Dutch guilders for male patients and 12,771 Dutch guilders for female patients (prices of 1996). Institutionalization is considerably more expensive: the average yearly cost associated with admission to a residential home is 48,180 Dutch guilders and 98,915 versus 102,930 Dutch guilders for costs associated with admission to a specialised respectively nonspecialized nursing home. Differences between men and women could not be taken into account in these last three estimations. The cost of living at home increases when the cost of informal care is included. However, when the cost were valued with a tariff of 29.50 guilders per hour, institutionalisation remains more expensive than living at home. This is also the case for severely demented patients, as the costs of institutions are also related to the level of care. PMID- 10816893 TI - [Professional expertise of higher level employees; age stereotyping in self assessments and supervisor ratings]. AB - In this contribution the relationship between higher level employees' age and two different types of assessments, i.e. self-ratings and supervisor ratings, of professional expertise is described. In research literature age stereotypes are often reported to influence supervisor ratings. Two hypotheses were tested with original survey data, collected in 1995, from 420 higher level employees and 224 of their direct supervisors. For the measurement of professional expertise, a domain-independent measurement instrument was developed consisting of five different scales, i.e. knowledge, meta-cognitive knowledge, skills, social recognition and growth and flexibility. The instrument consists of 78 items and is very reliable. The outcomes regarding discriminant and convergent validity indicate that the distinctive power of the five scales is satisfactory. Concerning the analyses of the effects of age, our hypotheses have largely been confirmed. We found that employee ratings do not differ by age. By and large, supervisors have a more positive view concerning the capabilities of younger employees. Our data indicate that supervisors make a difference in evaluating the higher level over-fifties compared with the evaluation of the middle-aged (35-49) and the starters (20-34). Pairwise comparisons indicate a leniency effect; in general self-ratings are higher compared to supervisor ratings, in particular in the seniors. PMID- 10816894 TI - [Memory tests in healthy elderly: age factors and norms]. AB - The study aimed to assess the strength of age effects on both standard laboratory and ecological memory tests and the psychometric qualities of these tests. Furthermore, norm data are constructed. Memory performance was assessed in a random group of older adults (mean age 62 years, range 46-89) and a group of older adults having memory complaints who applied for memory training (mean age 63 years, range 45-85). Age effects were found on almost all memory tests, whether artificial laboratory or more ecological tests were used. Age effects remained generally present after correction for educational level. Retest reliabilities of the ecological memory tests did not differ systematically from those of standard laboratory tests. However, not all tests showed satisfactory retest reliabilities, this was even true for tests often used in clinical settings. For tests with retest reliabilities above r = .65 norms corrected for age and educational level were provided. The group of older adults having memory complaints performed on average better than the random group of older adults from the population. In the first group, higher performance thresholds should be employed in assessing whether memory performance is deviant. PMID- 10816895 TI - [Clonidine within the scope of balanced inhalation anesthesia with sevoflurane- effects on pEEG parameters]. AB - The anaesthetic-saving property of clonidine has often been reported. In our own prospective, randomized study, in which the depth of anaesthesia was controlled by using spectral edge frequency (target-SEF90 = 10 Hz) and clinical parameters, we confirmed the anaesthetic-saving property only for fentanyl (-20%). On the other hand, there was no difference in MAC-sevoflurane values between the groups in keeping a steady target-SEF. For this reason, we analysed the stored pEEG data with regard to clonidine-specific effects. Twenty-eight patients were included in our study. The patients were randomly treated preoperatively with an infusion of 4 micrograms/kg KG Clonidine (clonidine group) or a placebo (placebo group). For anaesthesia, a standardized procedure with fentanyl, propofol, rocuronium, N2O/O2/sevoflurane was performed. The depth of anaesthesia was controlled by using SEF90 and clinical parameters. SEF90, SEF50 and the EEG power-spectrum were analysed over a five-minute period in the steady state of anaesthesia without surgical manipulation. Both placebo and the clonidine showed comparable values for SEF90 (9.9 +/- 1 vs. 10.7 +/- 0.9 Hz). On the other hand, there were differences in the SEF50 values between the groups (3.3 +/- 0.7 vs. 6.4 +/- 2.2 Hz). In the clonidine group, relative alpha-power was higher than in the placebo group (36.3 +/- 15.8 vs. 16.8 +/- 8.8%). Parallel to this effect, there was a reduction in relative delta-power in the clonidine group (47.6 +/- 15.2 vs. 65.4 +/- 9.1%). The described EEG effects on the power-spectrum inevitably influence SEF50 and SEF90. In our opinion, SEF50 is not a powerful predictor of depth of anaesthesia, when anaesthesia is performed in the way described. By performing this variation of balanced anaesthesia and co-medication with clonidine, higher values of SEF90 (11-14 Hz) seem to be adequate for surgical manipulation. PMID- 10816896 TI - [Initiation of lipid peroxidation (LPO) in blood during intraoperative mechanical autotransfusion--is hepatotoxicity of lipid peroxidation products of clinical significance?]. AB - Machine autotransfusion using cell-saver is a well-established method of saving homologous blood during extensive surgical procedures. The processing of blood may induce the initiation of lipid peroxidation (LPO) with the release of hepatotoxic products. A series of 42 patients undergoing primary (n = 20) or revision (n = 22) hip arthroplasty comprised the study group. Patients received an average of 1,260 ml of autologous blood and 2.2 units of homologous packed cells. The concentration of thiobarbituric acid reactive substances (TBARS) as LPO metabolites was measured in the patients' plasma, in the autologous packed cells as well as in the supernatants of the cell-saver-processed blood. Additionally, parameters of iron metabolism, haemoglobin levels, haematocrit as well as the activities of so-called liver enzymes aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltranspeptidase and cholinesterase were determined. An initiation of LPO was detectable during the process of machine autotransfusion, but this took place mainly ex vivo. High concentrations of TBARS were detectable in the supernatants after cell-separation processing. We observed a decline in haemoglobin concentration and haematocrit during the perioperative period. Postoperatively, we found a significant iron deficiency as a consequence of the perioperative blood loss. There was not sufficient evidence of a postoperative liver disorder induced by toxic metabolites of LPO. To sum up, there is only a low contamination of the organism with LPO products during the process of machine autotransfusion. Therefore, an induction of liver damage seems to be improbable. PMID- 10816897 TI - [How accurate is invasive blood pressure determination with fluid-filled pressure line systems?]. AB - It has been discussed by other authors that clinically relevant mistakes can occur in the measurement of invasive blood pressure. For this reason, we investigated all pressure lines used in our hospital. Our studies showed that exact measurement of invasive blood pressure using fluid-filled pressure measuring systems is not possible in the arterial, venous or pulmonal-arterial areas. Iatrogen mistakes should be excluded by working carefully. Exact knowledge of the physical qualities and the dynamic response of the fluid-filled pressure line used is required for judging the measuring accuracy. In clinical practice, measurement errors can amount to 40%. An acknowledged method to verifying errors is the Gabarith system developed by Billiet and Colardyn, which can determine the extent of the biggest-possible error after investigating the pressure line. A reduction of measurement error to below 2% can be achieved by carefully combining the individual components of the measuring system and, if necessary, by using an industrially-produced damping device (resonance overshoot eliminator [R.O.S.E.]). In this way, standardized measuring systems can be made available for clinical practice. PMID- 10816898 TI - Analysis of salbutamol and related impurities by derivative spectrometry. AB - Ultraviolet derivative spectrometry has been proposed for the analysis of salbutamol and related impurities. The assay of salbutamol aldehyde, 5-formyl saligenin, and salbutamol ketone was performed in sodium hydroxide 0.1 mol/l solutions, using first and second derivative spectra. The method has been applied for the assay of related impurities of commercial samples of salbutamol sulfate. PMID- 10816899 TI - Studies on some 3(2H)-pyridazinone derivatives with antinociceptive activity. AB - Nineteen new [6-(4-methoxyphenyl)-3(2H)-pyridazinone-2-yl]-acetamide (1-10) and 3 [6-(4-methoxyphenyl)-3(2H)-pyridazin-one-2-yl]propanamide (11-19) derivatives have been synthesized in this study. The structures of the compounds have been elucidated by their IR and NMR spectral data and elemental analysis. Antinociceptive activity of the compounds has been investigated by modified Koster's Test in mice, using aspirin as a reference. All the compounds (at 100 mg/kg dose) except 1 and 9 have been found more potent than aspirin. Compound 6 in the group of acetamide derivatives and compound 15 in the group of propanamides exhibited the highest antinociceptive activity. In addition, the propanamides have generally been found more potent than acetamides. In addition to these studies, the quantitative relationships between some structural parameters (such as log P, parachor, molar refractivity, and molecular connectivity indices) and antinociceptive activity of the compounds have been investigated. Statistical regression analysis has shown a close relationship to exist between the first-order molecular connectivity index (1 chi) and the antinociceptive activity. PMID- 10816900 TI - Naphthazarin derivatives (VI): synthesis, inhibitory effect on DNA topoisomerase I and antiproliferative activity of 2- or 6-(1-oxyiminoalkyl)-5,8-dimethoxy-1,4 naphthoquinones. AB - 2- or 6-(1-Hydroxyiminoalkyl)-5,8-dimethoxy-1,4-naphthoquin-one (DMNQ) and 6-(1 propyloxyimino- alkyl)-DMNQ derivatives were synthesized, and their inhibitory effects on DNA topoisomerase-I (TOPO-I) and antiproliferative activities against L1210 cells were examined. In a comparison, it was found that 6-(1 hydroxyiminoalkyl)-DMNQ derivatives exhibited higher potencies in both bioactivities than 2-(1-hydroxyiminoalkyl)-DMNQ analogues, suggesting that the difference in bioactivities between two positional isomers might be due to the steric hindrance of the side chain. It is noteworthy that the optimal size of alkyl group for both bioactivities of 6-(1-hydroxyiminoalkyl)-DMNQ derivatives was pentyl to octyl (IC50, 22-29 microM) for the inhibition of TOPO-I and propyl to nonyl (ED50, 0.12-0.19 microM) for the antiproliferative activity. In addition, a similar potency of bioactivities was expressed by 6-(1 propyloxyiminoalkyl)-DMNQ derivatives, propylation products of the oximes. PMID- 10816901 TI - Cytotoxic activity of cerium complexes with coumarin derivatives. Molecular modeling of the ligands. AB - Cerium complexes of Umbellipherone, Mendiaxon, Warfarin, Coumachlor, and Niffcoumar have been synthesized by reaction of the ligands with cerium nitrate in a stoichiometric ratio of 1:2. The formation of the complexes has been proved on the basis of elemental analysis, conductivities, IR spectroscopy, and 1H-NMR spectroscopy. The molecules of the ligands were optimized by means of the semiempirical quantum mechanical method PM3 to the energetically most stable conformers. All the ligands were characterized by molecular and submolecular electronic indices and the putative donor centers are proposed. It is concluded that the lactone- and the keto-carbonyl groups of Warfarin, Coumachlor, and Niffcoumar are bonded to the metal ion as bidentate ligands. The other two coumarins are bonded as monodentate ligands. Conductivity measurements show the non-electrolytic nature of the complexes. Cytotoxic screening by MTT assay was carried out. The cerium complexes were found to be more active than the inorganic salts. PMID- 10816902 TI - Synthesis, DNA-binding, and antiviral activity of certain pyrazolo[3,4 d]pyrimidine derivatives. AB - Some new pyrazolo[3,4-d]pyrimidine derivatives have been prepared and tested for their antiviral and DNA-binding activities. Compounds 4, 5, and 6 showed high binding affinity to DNA at concentrations of 19, 27, and 28 micrograms/ml, respectively. On the other hand, compounds 6 and 10 reduced the number of viral plaques of Herpes simplex type-1 (HSV-1) by 66 and 41%, respectively. The detailed synthesis and spectroscopic and biological data are reported. PMID- 10816903 TI - Health anxiety moderates the effects of distraction versus attention to pain. AB - Little is known about the relationship between health anxiety and chronic pain. The present study explored whether individual differences in health anxiety would influence the response of chronic pain patients to physical therapy. Furthermore, the interaction of health anxiety with coping strategy usage (distraction versus attention) was studied. Participants were 81 chronic pain patients who were interviewed and completed measures of pain, anxiety and cognition following an active physiotherapy session in which they either: (1) attended to physical sensations; (2) distracted from physical sensations or (3) completed the session as usual. Health anxious, compared to non-health anxious, individuals worried more about their health and injury during the session and attended to and catastrophically misinterpreted sensations more frequently. A complex interaction between health anxiety and coping strategy emerged. Among health anxious patients, attention to sensations resulted in lower anxiety and pain than did distraction. It appears as though attention had a short-term anxiety reducing effect for health anxious patients. Among non-health anxious patients, attention resulted in greater worry about health than distraction. Clinical and theoretical implications are discussed. PMID- 10816904 TI - Appraisal and control of sexual and non-sexual intrusive thoughts in university students. AB - This study examined differences in the appraisal and thought control strategies associated with the perceived control of unwanted sexual and non-sexual intrusive thoughts. Eleven appraisal dimensions, subjective physiological arousal and 10 thought control strategies were measured in 171 university students who were administered the Revised Obsessive Intrusions Inventory-Sex Version, a self report measure of unwanted intrusive thoughts. Thought-action fusion (TAF) likelihood was a significant unique predictor of the perceived controllability of respondents' most upsetting sexual and non-sexual intrusive thought. Moreover greater subjective physiological arousal was a significant predictor of reduced control over sexual intrusions, whereas worry that one might act on an intrusive thought and greater effort to control the intrusion were significant unique predictors of the control of non-sexual intrusive thoughts. Various thought control strategies were more often used in response to non-sexual than sexual cognitions. The results are discussed in terms of the differential role of various appraisal processes in the control of unwanted sexual and non-sexual thoughts. PMID- 10816905 TI - Worrying about chronic pain: vigilance to threat and misdirected problem solving. AB - Worry is reviewed to inform a cognitive-behavioural understanding of chronic pain. How worry comes to dominate is discussed as 'vigilance' and why worry comes to dominate is discussed as 'problem solving'. These two aspects are applied to worrying about chronic pain. Chronic pain is re-presented as chronic vigilance to threat that may lead to a perservation of attempts at solving the problem of achieving escape from pain. Attempts will be frustrated by the insolubility of the problem of chronic pain. The clinical and theoretical implications of this application are discussed. PMID- 10816906 TI - Child resiliency in inner-city families affected by HIV: the role of family variables. AB - This study examined the role of family variables in child resiliency within a sample of African-American, inner-city children whose mothers are HIV-infected. Variables from three dimensions of the family were included: family structural variables, maternal variables, and mother-child (parenting) variables. The participants were 82 children between the ages of 6 and 11 and their HIV-infected mothers. Correlational analyses indicated that resiliency was associated only with three parenting variables: parent-child relationship, parental monitoring, and parental structure in the home. Hierarchical regression analyses indicated a multiplicative relationship between parental monitoring and parent-child relationship and between parental monitoring and parental structure in the home, suggesting that parenting variables potentiate each other. Clinical implications of the findings are considered. PMID- 10816907 TI - Worry in children is related to perceived parental rearing and attachment. AB - In a sample of 159 primary school children, the relationship between perceived parental rearing behaviours and self-reported attachment style, on the one hand, and worry, on the other hand, was investigated. Children completed (a) the EMBU, a questionnaire measuring perceptions of parental rearing behaviours, (b) a single-item measure of attachment style, and (c) the Penn State Worry Questionnaire for Children (PSWQ-C), an index of severity of worrying. Results showed that parental rearing behaviours, in particular rejection and anxious rearing, were positively associated with worry. Thus, children who perceived their parents as more rejective and anxious reported higher levels of worry. Furthermore, self-reported attachment style appeared to be related to worry. More specifically, children who classified themselves as avoidantly or ambivalently attached displayed higher levels of worry than did children who classified themselves as securely attached. These findings are consistent with the notion that family environment factors such as parental rearing and attachment style contribute to the severity of anxiety symptoms in children. PMID- 10816908 TI - Predicting who suffers psychological trauma in the first year after a road accident. AB - This study followed up a cohort (n = 72) of consecutive road accident attendees to hospital for one year, assessing them soon after the accident and 3, 6 and 12 months later with the General Health Questionnaire (28-item version); Impact of Event Scale and Posttraumatic Stress Disorder Interview. The aim of the study was to identify demographic, accident and subjective reality variables that could predict who was likely to suffer psychological disorder in the future. The extent of injury was a particular focus for the study. Results showed that at different time periods, using stepwise, multiple regression analyses, between 12 and 77% of variance in trauma measures could be predicted. Severity of injury was a stronger predictor longer after the accident. PMID- 10816909 TI - Phobic dimensions: IV. The structure of animal fears. AB - Research designed to determine the number and kind of dimensions underlying self reports of animal fears is relatively rare. To contribute further knowledge to this area of study, Davey's methodology [Davey, G. C. L. (1994a). Self-reported fears to common indigenous animals in an adult UK population: the role of disgust sensitivity. British Journal of Psychology, 85, 541-554.] was improved. Principal components analysis with Varimax rotation of the self-ratings to items of Davey's Animal Fears Questionnaire returned by Ss from a Dutch community sample (N = 214) revealed four reliable, relatively independent dimensions: (1) fear-relevant animals, (2) dry or non-slimy invertebrates, (3) slimy or wet looking animals and (4) farm animals. Replicating Davey (1994a), females, relative to males, reported higher levels on most fear items. Principal components analysis with Oblimin rotation involving animal fears scales (derived from the dimensions identified in the present study), dimensions of non-animal fears, disgust sensitivity, sex-role orientation and the major dimensions of personality from the Eysenckian system revealed 4 higher-order factors, namely specific animals fears, positive affectivity, toughmindedness and negative affectivity. At an even higher level, these 4 higher-order factors merged into two factors: (1) a bipolar positive affectivity versus neuroticism/general emotionality/negative affectivity factor and (2) a toughmindedness dimension. Studies such as these contribute in helping provide the elements of the hierarchical model of fears proposed by Taylor [Taylor, S. (1998). The hierarchic structure of fears. Behaviour Research and Therapy, 36, 205-214.]. Findings across different studies suggest that there are at least 5 first-order dimensions of animal fears, the above 4 and predatory (fierce) animals, that may be included in such a model. PMID- 10816910 TI - [Anthelmintic resistance in gastrointestinal Strongylidae in sheep and goats: new data on prevalence, epidemiology, preventive measures and alternatives to anthelmintic drugs]. AB - Anthelmintic-resistant gastrointestinal nematodes represent the major problem in controlling of parasites of small ruminants in central Europe. This article gives a brief update on the prevalence of resistant populations in sheep and goat farms in Europe. New data on the selection and spread of anthelmintic resistance are reported and recommendations are given for minimising further increases in the incidence of resistant populations. New strategies which may play a role in integrated control programs in the future, e.g. vaccination and the development of nematode-resistant breeds of animals or of nematophagous fungi are briefly discussed. PMID- 10816911 TI - [Endoparasites in swine in different age groups and management systems]. AB - In different age groups endoparasites are of varying importance. Sows are predominantly infected with gastrointestinal strongyles (mainly Oesophagostomum spp.), Ascaris suum and Eimeriae, to a lesset extent with Trichuris suis, Hyostrongylus rubidus, Strongyloides ransomi or Isospora suis, while in suckling piglets I. suis and occasionally S. ransomi frequently occur. Weaners and fatteners are infected with coccidia and gastrointestinal strongyles, later with A. suum. Whipworms are also found occasionally. I suis causes diarrhoea in suckling piglets, while in older animals the infection is not correlated with disease. Damage induced by helminths is mainly due to reduced performance, in the case of ascarosis to reduced carcass value. Therefore planned antiparasitic measures should be taken at all stages of pig keeping. Hygienic measures can reduce infection pressure; however parasite elimination is difficult to obtain under conventional management conditions. Consequently the control of parasitic infection at the early stage of piglet production is desirable. PMID- 10816912 TI - [Current status of the discussion on the certification of so-called "Trichinella free areas"]. AB - In the last century already, Friedrich Albert Zenker and Rudolf Virchow have provided the essential prerequisites for the examination of pork for Trichinella in Germany. According to Directives 64/433/EEC and 77/96/EEC, this examination is still required in all member states of the European Union (EU) for trade between EU countries and for pork imported from third countries. According to Directive 64/433/EEC (Article 6, para 2), the examination of swine for Trichinella may be dispensed with if, on the basis of epidemiological studies, the stocks are found to be free from Trichinella and there is an efficient system of ante-mortem and post-mortem examination and control. However, it had been impossible to introduce these provisions into practice due to the fact that criteria were lacking. Therefore, the Scientific Veterinary Committee of the European Commission established a working group which has elaborated a concept for the certification of Trichinella-free pig stocks. The concept includes, inter alia, definitions of the essential components of "non-endemic area" and "Trichinella-free stock" and the requirement to be complied with for certification. Details are given on measures taken to control continued compliance with the certification requirements, the reporting and documentation system and the measures to be taken when trichinellosis is detected at a certified farm. PMID- 10816913 TI - [Control of Echinococcus multilocularis in Central Europe-- possibilities and limitations]. AB - Human alveolar echinococcosis is a life-threatening parasitic zoonosis which occurs in rare cases in Central Europe. It is caused by the larval stage of Echinococcus multilocularis. In this publication, strategies for the control of parasites of the genus Echinococcus are reviewed and discussed with regard to their applicability to E. multilocularis on the background of the epidemiological situation in Central Europe. First results of field studies are presented in which strategies based on the application of praziquantel-containing baits to foxes have been tried. Requirements concerning the surveillance of control measures are demonstrated and the limitations of the imaginable control strategies discussed. PMID- 10816914 TI - [Eradication of Chorioptes bovis mange in a dairy herd with turn-out to graze by Eprinex Pour-On]. AB - The possibility to control or even eradicate chorioptes manage by a single herd treatment with EPRINEX Pour-On (dosage: 0.5 mg Eprinomection/kg bodyweight) during pasture season was investigated in a dairy herd of 320 dairy cows. A further aim of the study was to evaluate whether such mid-summer treatment with EPRINEX Pour-On due to its endo-ectocide action would at the same time also result in a metaphylaxis of gastro-intestinal and lung worms and in a reduction of fly infestation. Due to clinical symptoms manage prevalence in the herd prior to treatment was 11 percent. After treatment, clinical symptoms disappeared within 2 months completely and did not re-occur during the subsequent housing period. Chorioptes mites during the entire trial period no longer were detected. The final clinical and parasitological investigation shortly before turn-out the next year (April 1999) demonstrated chorioptes mange to be eradicated clinically and parasitologically. Due to the mid-summer treatment infestation with gastrointestinal helminths also was eliminated and clinical symptoms of helminth infestations during the pasture season no longer were observed. However, at housing in November, low numbers of eggs of gastrointestinal nematodes were detected in 6% of dairy cows, 32% of second-season and 63% of first-season heifers, respectively. Milk yield per cow and day on average increased by 1 litre after treatment with EPRINEX Pour-On. This increase in production is likely a result of the reduction in total parasite burden of lactating cows. PMID- 10816915 TI - [Treatment of oestriasis and hypodermosis in red deer (Cervus elaphus hippelaphus) and roe deer (Capreolus c. capreolus) with ivermectin (Ivomec)]. AB - In two test areas in Austria (Burgenland, Lower Austria) ivermectin has been used to control oestrinosis and hypodermosis in red- and roe deer since many years (1985 and 1987, respectively). The results until 1992, using the injectable solution Ivomec-injection 1% only, have been reported (Kutzer, 1994). In continuation of these tests Ivomec-Praemix 0.6% was examined and it turned out that the treatment was extremely successful. In practice Ivomec-Praemix 0.6% has to be given preference due to its stability. The most favourable dosage for red- and roe deer was 2 x 0.4 mg ivermectin/kg bodyweight administered at an interval of one week, although equally good results could be obtained with 1 x 0.3-0.4 mg ivermectin/kg bodyweight. With the above mentioned dosages against Pharyngomyia picta and Cephenemyia stimulator an up to 100% and against Hypoderma actaeon a 100% success of treatment could be achieved. The administration in two doses has the advantage to eradicate intestinal and lung nematodes at the same time if there are any. Finally, it was found that the controlling measures should not be interrupted for more than one year, if no treatment was done in the neighbouring hunting grounds. PMID- 10816916 TI - [The flea in cultural history and first effects of its control]. AB - The knowledge about the biology and the origin of fleas from different periods of time as well as different geographical regions are described, starting from antiquity. Among this, the role of fleas in literature and art, for example in fairy tales and legends, proverbs and fables, painting and music and also the so called "circus of fleas" are covered. In connection with those aspects of cultural history the first efforts of control of flea plagues are mentioned. In addition to magic spells, these also include more or less complicated fleatraps for fleas and attempts of chemical control. PMID- 10816917 TI - Sedation agents. PMID- 10816918 TI - DGA discrepancies corrected. PMID- 10816919 TI - Pivotal points. PMID- 10816920 TI - Bacteraemia. PMID- 10816921 TI - Bacteraemia. PMID- 10816922 TI - Emergency dental drugs. PMID- 10816923 TI - Dental treatment for people with challenging behaviour: general anaesthesia or sedation? AB - The dental care of people with severe learning disability and challenging behaviour presents many problems. The maintenance of oral health by regular examination, prevention and treatment may be difficult because of the limitations in patient cooperation. In many cases the diagnosis of orofacial pain may need to be discounted as a cause of uncharacteristic and sometimes aggressive behaviour. In such cases the use of general anaesthesia for examination and treatment would seem to be the obvious option but this strategy has limitations. This paper undertakes a focused review of sedation techniques as an alternative to general anaesthesia in the treatment of people with challenging behaviour. The use of novel techniques of sedation combining intravenous with oral or intranasal routes is described with patients treated in a community dental health centre. All patients had previously received treatment using general anaesthesia. The techniques described proved effective and safe for use in the primary care setting. PMID- 10816924 TI - Oral cancer in young adults: report of three cases and review of the literature. AB - Oral cancer in young adults is fortunately uncommon in the UK. However, since it is so rare, when cases present they are often misdiagnosed and inappropriately treated leading to delay in definitive treatment. This may, in turn, lead to a poorer prognosis for these patients. It is debatable if oral cancer in younger adults carries an inherently poor prognosis and presents with more aggressive tumours. Three cases of oral cancer in young adults, aged under 30 years are presented and the literature reviewed with respect to oral cancer in this group of patients. PMID- 10816925 TI - Comment on: oral cancer in young adults. PMID- 10816926 TI - Interesting presentation of a retained upper deciduous incisor with apical fenestration. AB - A case is described where intrusion of a primary incisor resulting in apical fenestration may have occurred secondary to occlusal forces. The effects on the permanent dentition and possible aetiological factors are discussed. PMID- 10816927 TI - Complete dentures: an introduction. PMID- 10816928 TI - Clinical assessment. PMID- 10816929 TI - Referral patterns and the referral system for oral surgery care. Part 2: The referral system and telemedicine. AB - OBJECTIVE: To investigate GDP opinions of the current referral system and to investigate the need and demand for telemedicine in oral surgery referrals. DESIGN: Postal questionnaire. SETTING: 400 GDPs in Greater Manchester. RESULTS: 84% participation rate. 48% were not satisfied overall with the service of their current specialist oral surgery referral site. The principal reason was the length of the waiting time for consultation and treatment. Distance for patients to travel to the specialist unit was also of concern, even though most patients (89%) travelled short distances (return journey of twelve miles or less). 23% of respondents wished to improve their ability to communicate with the oral surgeon and 70% wanted involvement in the patient consultation. Both of these requirements were more likely in younger practitioners. CONCLUSION: There is a need and demand for change in the referral system for oral surgery specialist care. Telemedicine could conceivably be one way to improve access to specialist oral surgery care. PMID- 10816930 TI - The effect of orthodontic referral guidelines: a randomised controlled trial. AB - OBJECTIVE: To develop and evaluate the effectiveness of referral guidelines for the referral of orthodontic patients to consultant and specialist practitioner orthodontists. DESIGN: Single centre randomised controlled trial with random allocation of referral guidelines for orthodontic treatment to general dental practitioners. SETTING: Hospital orthodontic departments and specialist orthodontic practices in Manchester and Stockport. SUBJECTS: General dental practitioners and the patients they referred for orthodontic treatment. MAIN OUTCOME MEASURE: Appropriateness of referral, defined as whether the patient was accepted for orthodontic treatment. RESULTS: The referral guidelines did not reduce the number of inappropriate referrals. CONCLUSIONS: Referral guidelines for orthodontic referrals did not influence the behaviour of the general dental practitioners. More research into the optimum methods of dissemination and implementation of referral guidelines for use in the general dental service is needed. PMID- 10816931 TI - Antibiotic prescribing for dental conditions: general medical practitioners and dentists compared. AB - OBJECTIVE: To compare the level of prescribing and types of antibiotics prescribed for dental problems by general medical practitioners and dentists. DESIGN: Secondary analysis of standard consultation data and prescription records from four different settings. SETTING: 30 participating general practices in the General Practice Morbidity Database for Wales in 1996. SUBJECTS: 1,185 attendances for tooth-related problems, at 30 participating practices in the General Practice Morbidity Database for Wales in 1996. Standard consultation records from a GDP emergency dental rota, and two weekend emergency dental clinics: one in a health centre, the other in a dental hospital. RESULTS: More than two thirds (68%) of attendances at general medical practices for tooth related problems resulted in a prescription for antibiotics. In contrast less than a third (28%) of patients seen by a GDP rota, about half (52%) of patients at a weekend emergency clinic in a health centre, and just more than a third (38%) of patients attending the dental hospital clinic received antibiotics. General medical practitioners were also more likely to prescribe broad-spectrum antibiotics than dentists. CONCLUSION: For acute dental problems general medical practitioners are more likely to prescribe antibiotics than dentists. There also appear to be inter-professional differences in the tendency to prescribe broad spectrum antibiotics. Initiatives to rationalise prescribing for dental conditions may need to target doctors as well as dentists. PMID- 10816932 TI - [The cerebral organization of creativity. I. The development of a psychological test]. PMID- 10816933 TI - [A neuropsychological approach to the analysis and evaluation of visual representations]. PMID- 10816934 TI - [Evoked cortical potentials of the cerebral hemispheres during a comparison of visual stimuli]. PMID- 10816935 TI - [The specialization of the cerebral hemispheres in the perception of the intonation of the Russian language]. PMID- 10816936 TI - [The comparative characteristics of spatial hearing in patients with different forms of cortical epilepsy]. PMID- 10816937 TI - [The EEG parameter dynamics of younger schoolchildren during a functional change in CNS status (relaxation) under exposure to rhythmic auditory stimulation]. PMID- 10816938 TI - [The dynamics of cardiac rhythm during sleep in young infants with the perception of tonal stimuli]. PMID- 10816939 TI - [The characteristics of the personality component in the psychophysiological potential of different socio-professional groups]. PMID- 10816940 TI - [The dynamic characteristics of the human resonance EEG reactions to rhythmic photostimulation]. PMID- 10816941 TI - [Jendrassik's maneuver creates the conditions for triggering involuntary stepping movements]. PMID- 10816942 TI - [The effect of general cooling on the electromyographic characteristics of muscle fatigue evoked by dynamic loading]. PMID- 10816943 TI - [The individual somatotype characteristics and skeletal muscle energetics of girls 7 to 11 years old]. PMID- 10816944 TI - [The characteristics of the morphological traits and body function in adolescents adapting to the North]. PMID- 10816945 TI - [Eiconol, a complex of unsaturated fatty acids, optimizes human cognitive activities]. PMID- 10816946 TI - [The phospholipid profile of inhabitants of the European North of Russia]. PMID- 10816947 TI - [The estimation of a complex of hemorheological parameters in erythrocytosis]. PMID- 10816948 TI - [A prognostic factor for the risk of developing pathological processes based on the polymorphism of the enzymes of xenobiotic metabolism]. PMID- 10816949 TI - [A hypothesis on the nature of electrodermal reactions]. PMID- 10816951 TI - [In Process Citation] PMID- 10816950 TI - [The immunological reactivity of healthy persons of different ages and the ABO system antigens]. PMID- 10816952 TI - [The axiologic structure of the human]. AB - The essential characteristic of human being demands unveiling the variety of values that constitute it and create a specific "axiological structure". The main features of this structure are as follows: 1. the axiological structure of human being is radically individualistic; 2. the above structure comprises different types of values (not only strictly moral ones); 3. the structure in question is hierarchical; 4. the axiological structure comprises both positive and negative values; 5. the axiological structure contains within itself both deterministic and indeterministic factors; 6. the axiological structure is dynamic; 7. the axiological structure is dialectic; 8. the axiological structure is made up of both the actually existing and potential values, as well as the realized and merely postulated values; 9. the axiological structure may be characterized by both the harmony and disharmony of its component elements (values); 10. the axiological structure has a teleological character: its most significant "determining" factor is man's attitude towards values. PMID- 10816953 TI - [Quality of life: an ethical and medical issue]. AB - Quality of life as a criterion of evaluation of treatment and other medical activities was analysed with respect to meaning and the differences which occur in the understanding of quality of life by the physician and the patient. Attaining has been paid to these conditions under which the opinions of the physician and patient are common. It is proposed to accept the value of life as superior to quality of life. Realising such principle the possibility of compatibility of moral and pragmatic arguments could be expected in the use of quality of life as a criterion. PMID- 10816954 TI - [Is personalism or utilitarianism an adequate foundation of medical ethics?]. AB - The article rejects utilitarianism as a proper theory for medical ethics. Utilitarians lavishly use various slogans of effective action, development and better civilization. However, the principle of prosperity of humanity in the utilitarian interpretation makes the value of the human person subject to society. Social interest threatens the individual here because it defines his/her value of life. The drift towards maximalization of benefits and prosperity of humanity strikes the seriously ill, e.g. babies with brain damages, Down's syndrome, etc., people after accidents and with serious brain defects, the terminally ill. The principle of quality of life (lebensunwertes Leben) used by utylitarians allows them to argue, that euthanasia, abortion is in the interest of the patient. Some utilitarians openly admit that such ideas as "universal happiness", "prosperity", "benefit" are empty ideas, fictions to which one cannot attribute any contents. So utilitarianism, not defining its fundamental ideas, can easily change medical ethics in a theory of elimination of the uncomfortable people. Therefore, as a theory utilitarianism cannot serve as the basis for medical ethics. PMID- 10816955 TI - [Teachings in bioethics from the Pontifical Academy of Theology in Cracow]. AB - The paper consists of two parts. In part one the author outlines conditions of introducing bioethics as lectures in university faculties of medicine and theology. After a few years of intensive development of bioethics, now some stagnancy can be observed. The author analyzes reasons for slowing down the intensity of that development. In part two the author reports of teaching activities in the Institute of Bioethics of the Pontifical Academy of Theology in Cracow. The teaching is on two levels. There is a basic course and postgraduate studies for theological and philosophical students. Also courses and conferences for health care workers are held. PMID- 10816956 TI - [The psychology of suffering]. AB - Man's common experience interpreted in the humanities points to the inevitability of suffering. Suffering was even termed a basic border situation (C. Jaspers). The so-called scientific psychology hardly analyses human suffering. S. Freud regarded culture as a source of suffering and the behaviourists did not see the need for an analysis of man's inner experience. Radical change of the view on suffering was brought about in the works of V. E. Frankl who thought man to be homo patients (to be a man means to suffer). There are various sources and kinds of suffering. Based on his own vast researches the author characterizes three kinds of man's suffering: physical (pain, somatic diseases), psychical (hardships, mental disorders and illnesses) and spiritual (lack of a meaningful life, moral dilemmas). He also puts forward a hypothesis that a psychic suffering is a correlative between both mental and spiritual sufferings. Suffering fulfills a variety of functions in the life and personality of a human being; it can cause a personality degradation or can further a personality development. Therefore, we can speak of the ambivalent character of suffering. The elementary psychological problem, encountered in suffering, is to give suffering some meaning. In order to do this one must ask about future: for whom and what do I suffer? Whereas the question about the ultimate origin of suffering (why do I suffer?) not only makes it possible to explain the issue fully but first of all allows us to recognize suffering as a mystery of human existence. The author proves that the analyses carried out by John Paul II in Salvifici doloris are coherent with the principles of logotherapy. However, the analyses are more profound since they point to individual, social, cultural and transcendental dimensions of suffering. PMID- 10816957 TI - [Medical ethics and patient responsibility]. AB - When we say "medical ethics" we understand the responsibility of the physician for his medical education and his attitude to his patient. But Hippocrates is known to have said that the efficiency and good results of the treatment depends not only on the physician but on the patient and his engagement, his observance of the doctors' advice, his attitude to his own psyche and body, both in health as in illness. This is an ethical problem known to every practitioner, the problem of ethics of the patient, which ought to be more widely disseminated in society. PMID- 10816958 TI - [Persistent vegetative state: medical, moral, legal and economic aspects]. AB - The classical definition of persistent vegetative state describes patient who is wakeful but not awake, and indicates intact function of the brain stem and no function of brain cortex. The presence and the degree of awareness is most difficult to assess and causes moral and legal controversies concerning the management of persistent vegetative state. The absence of the perceptual awareness may be the most important clinical characterization for diagnosis. Differential diagnosis includes coma, brain death, locked-in syndrome and minimally responsive state. The pressure of economical factors (managed care) on medical care makes this problem even more complicated. PMID- 10816959 TI - [Ethical aspects of preimplantation diagnostics]. AB - Pre-implantation Genetic Diagnosis (PGD) enables to genetic examination of an embryo springed up in vitro before his transfer to mother's womb. The ethical valuation of this affair is different. Fundamentally, ethic-medical debate takes the interest in practical advantages and in unwholesome of this method. But ethic theological discussion indicates' that PGD is the breaking of regulations concerning in the protection of the embryo. According to theological point of view, PGD is also the violation of conviction about human dignity and his rights to life from the conceiving. Therefore human embryo from his beginning should be protected against the instrumentalization or the extermination. Moreover the selection of embryos supposes a repartition between valuable and worthless life. Totally, PGD is contradictory to the teaching of Church. It produces greater complications than solutions of problems. PMID- 10816960 TI - [Histopathologic consultations for quality control. Ethical aspects]. AB - Histopathology and cytopathology are subjective, descriptive methods and there is a need for second opinion of second pathologist. This should be solved in a systematic way from both, organisational and financial point o view. This could not rely on a friendly consultation of a colleague pathologist only. Consultations are integral part of quality assurance and quality control. Quality control in gynaecological cytopathology is associated with reexamination of 1 in every 100 negative cytological smears. High grade dysplasia of the gastric mucosa requires rapid second endoscopy and second opinion prior to gastrectomy. All this is discussed from the point of view of common sense and ethics which (thanks God) are compatible. Protection against unwanted genetic molecular tests still is not satisfactory. PMID- 10816961 TI - [Physician-patient relations: sympathy or empathy?]. AB - At the beginning the author presents the definitions of antipathy, sympathy, empathy and syntony as different types of human interrelations. Then he recalls the kinds of physician-patient relations and discusses the antipathy, especially sympathy and empathy appearing in those relations. At the end the author proves the superiority of empathy over sympathy in the solution of problems of medical practice to the advantage of both the patients and the physicians. PMID- 10816962 TI - [[Selected legal aspects related to medical practice]. AB - The question of the physician's liability, both that of civil as well as penal law nature--is always emotionally approached. Dynamic development of medical and biological sciences as well as technics is the cause of progress but it also gives rise to the increase of hazards or abuses in medical therapy. If we speak of the therapeutic intervention being originally legal we mean that it is carried out in compliance with the principles of medical art. In such circumstances, even though the intervention resulted in negative effects, the intervening physician cannot be made penally liable. Civil law liability, in its turn, may have either ex contractu or ex delictu basis. When the general prerequisites of this kind of liability are present, the intervening physician (Art. 353 or 415 of Civil Code) or the State Treasury (Art. 417 of Civil Code) may be made liable for causing damage, joint and several liability of the physician and the Treasury being also possible (Art. 420 of Civil Code). The carrying out of therapeutic intervention without the law required consent of the patient may lead--on the basis of Polish law--to the physician's civil law liability for the infringement of the patient's personal interests even though the intervention ended in success (Articles 23 and 24 of Civil Code). From the point of view of Polish penal law such situation may cause the physician's penal liability for the offence against freedom (Art. 192 of Penal Code). The euthanatic homicide should be, and in Polish law, is an offence. Considering the potential abuses arising from making the euthanasia legal, penal law whose major function is that of the guarantee nature, must ensure safeguards vis-a-vis life to the utmost limit. Polish Legislator shows, however, full understanding of the extremely difficult and conflict-generating situation in which the individual committing euthanatic homicide may find himself. Hence, in section 2 of Art. 150 of Penal Code the Legislator declared that "in exceptional, particularly justified cases, the Court may apply extraordinary mitigation of penalty or even depart at all from meeting out the penalty". Law regulations cannot however solve problems whose moral and ethical dimension exceeds sometimes that limited to law only. Hence in plenty of cases the physicians are left to themselves with the "verdicts" produced by their own conscience. And indeed, these verdicts may many a time be more severe than the decision of the Court because the physicians cannot appeal from them. PMID- 10816963 TI - [Ethical aspects of forensic psychiatry and psychology]. AB - Psychiatry is a discipline which poses particularly sophisticated and subtile requirements from the point of view of professional ethics, as well as general morality, especially when a psychiatrists is a forensic court expert. The necessity of freeing oneself from the temptation to succum to one's distance, and preserve a suitable professional perspectives, with a simultaneous understanding of another human being, often forces the forensic expert to face the toughest and doctor. One also has to bear in mind the specific triple responsibility of the forensic psychiatrists: before the examined person, the confidence in him and whose interests also need to be respected and protected. Another deontological problem is the issue of keeping professional secrets which should belong to the duties of every psychiatrists and psychologists, and particularly that of a forensic expert. The issue of professional secrets in medicine is discussed in the Code of Medical Ethics (1993) and in Act on the Protection of Mental Health (1995). Finally, the deontological problems of forensic psychiatry and psychology can be analyzed at the most general level, when for instance, totalitarian states abuse institutions whose aim is to bring assistance to people with mental disturbances, for political purposes. Such accusation were never made with relation to Poland. PMID- 10816964 TI - [Gene therapy--hopes and fears]. AB - Gene therapy assumes the correction of a genetic defect by the delivery of a correct DNA sequence to the target cells. Depending on the target cells two types gene therapy have been defined: somatic and germinal. By July 1998, 351 protocols of somatic therapy were approved by the Recombinant DNA Advisory Committee. The majority of protocols focus on cancer therapy and monogenic diseases. By now, still there is more unfulfilled expectation than clinically sound achievements, since no effective prevention or successful treatment for genetic diseases or cancer have been developed. Germline genetic modification is considered as the treatment of choice for such a diseases like retinoblastoma. Tay-Sachs, Lesch Nyhan and metachromatic leuko-dystrophy. This approach which is still illegal or prohibited by rules in many European countries, is gathering more and more advocates. Once we learn how to control gene expression the perspectives for clinical application of gene therapy might be enormous. The safety of genetic modification of gametes or embryonal stem cells remains to be properly addressed and successfully solved. The ethical issues of germinal gene therapy are still the subject of controversial opinions among the scientists, lawyers and philosophers. PMID- 10816965 TI - [Personalization concepts of medical ethics in the teachings of John Paul II]. AB - According to John Paul's teaching, the medicine cannot be purely technological, but needs to have some human character. The medicine doctors should consider patients as a whole, not only their problems bound with the treating them as patients, and to cure them from their illnesses, but together considering their feelings, and their intellectual and spiritual life. John Paul's teaching in relation to "the personalizing of medicine" means such a consideration of the human being that is leading to the respect about the body, spirit and their culture. Therefore the knowledge about the biological properties of human life, should be accompanied together with the look onto their dignity. Nobody could be a doctor who cures only one organ of the human's body, but must see the whole human person together with the relations to other persons connected with that patient, because they have very often much of influence on to the health of a patient. That "personalizing of the medicine" should be done on such a way, when it is seen the total ill person. The doctor should be having friendly and even heartily personal relationship to the patient, which should lead to a dialog between them, full of respect and confidence. Nobody is allowed to consider an ill person as a clinical case, but should see a person who is expecting not only professional, competent medical attention, but also the understanding of the patient's spirit, which is being bound with that illness. In this way, John Paul II is telling us, that the health service should not be based entirely on the professional knowledge and skill. The doctor should see in the ill person his own dearest brother or sister, whom we are to love very much, and should be taking part in the illnesses of them. PMID- 10816966 TI - [The structure of meta-ethical reasoning and "paradigmatic instability" of medicine]. AB - Meta-ethics is such a philosophical discipline that it analyses meaning and logical status of moral conceptions and arguments. Its scope of interest is focused on such problems, among others, as to what extent are moral values the intrinsic part of our reality and, on the other hand, to what extent are they merely the expression or certain conventions, emotions or directives. Codes of medical ethics and other legal acts concerning ethical commitments, both domestic and international (as, for example, Declaration of Helsinki), are based on a certain hidden meta-ethical presumption that morality is a human creation, and thus it belongs to a certain so-called "social realm". As such, medical ethics undergoes continuous transformations with regard to technological progress, which creates new moral challenges. This frequently leads to conflicts between moral, legal and praxiological norms under the absence of consensual conformity of utilitarian and deontological ways of moral reasoning. It may be useful for medical ethics to differentiate between axiological and thetic norms that are present in codes of medical ethics and in various international bioethical declarations and conventions. PMID- 10816967 TI - [Definition of mental illness and discoursive strategies in psychiatry]. AB - Defining mental illness was presented in the article both as a matter of medical knowledge and a political issue. This latter aspect cannot be successfully dealt with by psychiatry itself, since it is a branch of medicine, nevertheless bioethics offers here its competences and possibilities. The presentation of some elements of traditional strategies in defining mental illness introduces a draft of such a project of the definition procedure, which reinforces the constantly threatened (by the decrease of sovereignity) social and legal status of psychiatry, and--on the other hand--enables us to support the evidently handicapped status of psychiatric patients. This solitary definition strategy, which support both psychiatric circles and patients, assumes that a popular modern tendency to deny the very reality of the mental illness is to be avoided. The definition of mental illness proposed in the article is pragmatic in character and is based on a definition of mental illness as a kind of spiritual disorder. PMID- 10816968 TI - [Bioethics in pluralistic societies--what does it all mean?]. AB - The phrase "bioethics in pluralistic society" has recently become a common phrase. At the same time it has become so notoriously ambiguous that it is worth to ask about its various meanings in order to reveal different senses and non senses hidden beneath it. The terms that compose this phrase have undergone critical analysis with particular emphasis put on the term "bioethics". The need for further development of bioethics as philosophical discipline, and not its various politically determined mutations, is stressed in concluding remarks. PMID- 10816969 TI - [The significance of animals in biomedical research]. AB - The mission of medicine is maintenance of health, elimination of suffering and prolongation of life. These aims can be achieved by medicine based on experimental determination, because only then it becomes a real science. The nature of human mind has led the man since the beginning of humanity on the earth to the cognition of his environment and himself. Being intellectually superior than other living creatures, the man got power over them. In his endless efforts to expand knowledge about living organisms, including his own, he started to use animals. The man has used animals for cognitive purposes for ages and is still doing it, however his motivation has changed and is still changing. Cognition of functions of living organisms on the basis of observation solely, without any interference into the living body gave a lot of important information, yet, generally, this method was of little use for the development of science. Only the use of animals could give information about this what was earlier unknown and impossible. The long-lasting evolution of experimental studies of living functions of higher organisms resulted in achieving a perfect level in biomedical studies. Vivisection, as it was understood years ago, has become history. For a chronic experiment, an animal is surgically prepared according to the researcher's intention. The surgery and the postoperative period follow the principles used in human surgery. After the convalesce period, the animal is used for further experiments. On such prepared animals, the investigations in experimental cardiology, neurophysiology, gastroenterology and other medical disciplines are performed. The animal prepared for longlasting experiments do not suffer from pain during both the experiments and intervals between them. Another important achievement in chronic experiments is considerable reduction of the number of animals used in experimental medicine. Undoubtedly, the greatest achievements in medicine in the 19th and 20th centuries were possible due to the use of animals. There is a strong relationship between a rapid progress in experiments on animals and evident progress in clinical medicine. In the second half of the 20th century the man left the globe for the first time and reached another planet. Tis unusual event in the history of humanity was possible due to space medicine based on biomedical experiments with the use of animals. Also, the man's ability to reach the depths of oceans cost the lives of many experimental animals. Despite advances that have already been made, we are still in the early stages of understanding the complex workings of the body. This makes the replacement of animal experiments a slow process. At the same time, our increasing knowledge is opening up whole new areas of medical research which in turn give rise to a need for further animal use. While it may be difficult to envisage total replacement, the proposition of work that can be done without animals is increasing all the time. Many efforts are being made to find alternatives to animal experiments, to increase the usefulness of those that already exist, and to refine animal research methods. But at present days biomedicine, with its experiments on animals, reveals the laws of nature which the clinical and his patient can use to improve the life quality, prolong the life span and eliminate suffering. We all want to lead a healthy and enjoyable life. Most of us want the benefits of modern medical research-benefits that we would not have without the contribution of animal research. Yet, the health and well being of animals is important too. Clearly, those involved in this research have a moral and legal obligation to ensure that laboratory animals are well housed and cared for, any distress is kept to a minimum and that as few animals as possible are used. In the end, the question of whether animals should be used in medicines research is one everyone must answer for themselves, but the decision is more c PMID- 10816970 TI - Genome complexity and organization in the red imported fire ant Solenopsis invicta Buren. AB - The red imported fire ant Solenopsis invicta is the most destructive invading arthropod in the southern United States, yet little is known about its genome complexity and organization. Here we report the size, organization and GC content of S. invicta genome. DNA reassociation kinetics using S1 nuclease assay and a modified second-order kinetics model indicated that the S. invicta genome is approximately 0.62 picograms or 5.91 x 10(8) base pairs, composed of 36% unique, 41% moderately repetitive and 23% highly repetitive/foldback sequences. Comparison of the reassociation kinetics of short and long DNA fragments revealed that the sequence arrangement follows a pattern of short period interspersion, as in most organisms with relatively large genomes. Melting-temperature analysis showed that the GC content of the fire ant genomic DNA is 34.8%, similar to that of most eukaryotic organisms. The results reveal that the fire ant genome is much larger and more complex than those of a number of hymenopteran insects studied to date. Our study provides a foundation for further analysis and genetic manipulation of the S. invicta genome. PMID- 10816971 TI - Kruppel homolog (Kr h) is a dosage-dependent modifier of gene expression in Drosophila. AB - A lethal mutation in the Kruppel homolog (Kr h) was isolated in screens of P element insertion mutations for modifiers of white gene expression. The mutation occurs in the 5' untranslated region of the Kr h gene and causes a lightening of the eye colour for several alleles of white due to a decrease in white steady state mRNA levels at pupal stages. Two related genes, scarlet and brown, were significantly affected as well in early pupae. Genetic analysis of different white alleles suggests that enhancer sequences are necessary for interaction with KR H. Thus, the Kr h gene is a member of the dosage-dependent hierarchy effective upon white. PMID- 10816972 TI - Chromosomal effects on male and female components of sperm precedence in Drosophila. AB - Recent experiments with Drosophila have demonstrated that the success of sperm in multiply mated females depends on the genotype of both the male and the female. To further characterize the distinction between male and female roles in sperm success, we scored variation in both sexes in sperm competitive ability among a set of chromosome replacement lines that allow identification of effects to each chromosome. We detected significant male and female effects on sperm precedence, defined as the ability of a male's ejaculate to displace resident sperm (P2) or avoid being displaced by subsequent matings (P1). Tests of effects of first, second and third chromosome substitutions revealed significant differences among third chromosomes in male sperm precedence (both P1 and P2) and a first x second chromosome interaction in female's effect on sperm precedence (only P1). We found no significant correlation between male and female effects on sperm precedence, suggesting that the variation found in both P1 and P2 has a different genetic cause in the two sexes. PMID- 10816973 TI - The genetic structure of female life history in D. melanogaster: comparisons among populations. AB - Two questions were addressed: (1) What is the genetic variance-covariance structure of a suite of four female life history traits in D. melanogaster? and (2) Does the genetic architecture of these traits differ among populations? Three populations of D. melanogaster were studied. Genetic variances and covariances were estimated by sib analysis three times for each population: immediately upon establishment of populations in the laboratory, and subsequently after approximately 6 months and 2 years of laboratory culture. Entire genetic variance covariance matrices, as well as their individual components, were compared between populations by means of likelihood ratio tests. All traits studied were significantly heritable in at least one-half of estimates. Despite large sample sizes, additive genetic covariances were for the most part not statistically significant, and only two significant negative covariance estimates were obtained throughout the experiments. Therefore, these experiments provide little support for evolutionary life history theories that are based on negative genetic correlations among life history components. Neither do they support the idea that genetic variance for fitness components is maintained by trade-offs. Evidence suggests that the G matrix of one population was initially different from those of the other two populations. Those differences disappeared after 2 years of laboratory culture. At the level of individual (co)variance components, there were relatively few differences among populations, and the overall impression was that the three populations had generally similar genetic architectures for the traits studied. PMID- 10816974 TI - Bayesian estimation of range for microsatellite loci. AB - Microsatellite loci have become important in population genetics because of their high level of polymorphism in natural populations, very frequent occurrence throughout the genome, and apparently high mutation rate. Observed repeat numbers (alleles size) in natural populations and expectations based on computer simulations suggest that the range of repeat numbers at a microsatellite locus is restricted. This range is a key parameter that should be properly estimated in order to proceed with calculations of divergence times in phylogenetic studies and to better investigate the within- and between-population variability. The 'plug-in' estimate of range based on the minimum and maximum value observed in a sample is not satisfactory because of the relatively large number of alleles in comparison with typical sample sizes. In this paper, a set of data from 30 dinucleotide microsatellite loci is analysed under the assumption of independence among loci. Bayesian inference on range for one locus is obtained by assuming that constraints on range values exist as sharp bounds. Closed-form calculations and robustness revealed by our analysis suggest that the proposed Bayesian approach might be routinely used by researchers to classify microsatellite loci according to the estimated value of their allelic range. PMID- 10816975 TI - The effect of epistasis on the structure of hybrid zones. AB - Within hybrid zones that are maintained by a balance between selection and dispersal, linkage disequilibrium is generated by the mixing of divergent populations. This linkage disequilibrium causes selection on each locus to act on all other loci, thereby steepening clines, and generating a barrier to gene flow. Diffusion models predict simple relations between the strength of linkage disequilibrium and the dispersal rate, sigma, and between the barrier to gene flow, B, and the reduction in mean fitness, W. The aim of this paper is to test the accuracy of these predictions by comparison with an exact deterministic model of unlinked loci (r = 0.5). Disruptive selection acts on the proportion of alleles from the parental populations (p,q): W = exp[-S(4pq)beta], such that the least fit genotype has fitness e-s. Where beta << 1, fitness is reduced for a wide range of intermediate genotypes; where beta >> 1, fitness is only reduced for those genotypes close to p = 0.5. Even with strong epistasis, linkage disequilibria are close to sigma 2p'ip'j/rij, where p'i, p'j are the gradients in allele frequency at loci i, j. The barrier to gene flow, which is reflected in the steepening of neutral clines, is given by [formula: see text] where r, the harmonic mean recombination rate between the neural and selected loci, is here 0.5. This is a close approximation for weak selection, but underestimates B for strong selection. The barrier is stronger for small beta, because hybrid fitness is then reduced over a wider range of p. The widths of the selected clines are harder to predict: though simple approximations are accurate for beta = 1, they become inaccurate for extreme beta because, then, fitness changes sharply with p. Estimates of gene number, made from neutral clines on the assumption that selection acts against heterozygotes, are accurate for weak selection when beta = 1; however, for strong selection, gene number is overestimated. For beta > 1, gene number is systematically overestimated and, conversely, when beta < 1, it is underestimated. PMID- 10816976 TI - Correlated responses to selection for large body size in oMt1a-oGH transgenic mice: reproductive traits. AB - Correlated responses in female reproductive performance were evaluated following short-term selection within full-sib families for increased 8-week body weight in two replicates of four lines of mice: two ovine metallothionein-ovine growth hormone (oMt1a-oGH) transgene-carrier lines, one from a high-growth background (TM) and one from a control background (TC), and two non-transgenic lines, one from each of these genetic backgrounds (NM and NC, respectively). A fifth line (CC), not containing the transgene, served as a randomly selected control. The initial frequency of the oMt1a-oGH transgene construct in the TM and TC lines was 0.5. The frequency of transgenic females sampled at generations 7 and 8 of selection was 84.0% and 6.1% in the TC and TM lines, respectively. No significant female infertility differences were detected between transgene-carrier and non transgenic lines or between transgenic and non-transgenic mice within carrier lines, whereas high-growth background lines had a higher infertility than control background lines (P < 0.05). Correlated responses in the TC transgene-carrier line were suggestive of reduced reproductive performance as indicated by increased post-implantation mortality (P < 0.05), number of dead fetuses plus implants (P < 0.05), and loss of fetuses from day 16 to parturition (P < 0.001). For the first two traits, the negative correlated responses were accounted for by the reduced performance of transgenic compared with non-transgenic females. Embryos carrying the transgene may also have a lower viability. In contrast, the NC non-transgenic line did not exhibit reduced reproductive performance for these traits. The low frequency of the transgene in the high-growth background TM line was associated with reduced fitness and a lower additive effect for 8-week body weight compared with the control background TC line. PMID- 10816977 TI - Serum leptin concentration in pigs selected for high or low daily food intake. AB - Selection for high or low daily food intake (DFI) in Large White pigs resulted in higher serum leptin concentration, fat deposition and food intake in the high DFI line. The response in serum leptin concentration indicated that the higher fat deposition of the high DFI line was not due to insufficient leptin production, as in the Lepob/Lepob mouse. Serum leptin was more highly correlated with fat deposition than with food intake indicating that the response in serum leptin was primarily due to increased fat deposition rather than to higher energy intake per se. The low correlations between serum leptin measured at 30 kg and performance test traits indicate that serum leptin would not be efficient for selection of animals prior to performance test. However, the consistent positive correlations between serum leptin and a measure of fat deposition suggest that serum leptin could usefully be incorporated in selection criteria for genetic improvement of carcass lean content in pigs. PMID- 10816978 TI - Quantitative genetic variation of leaf size and shape in a mixed diploid and triploid population of Populus. AB - In the interspecific cross of Populus trichocarpa x P. deltoides, unexpected simultaneous occurrence of diploid hybrids and triploid hybrids (with two alleles from the female parent and one from the male parent at each locus) led us to examine the evolutionary genetic significance of this phenomenon. As expected, leaf size and shape of the triploid progeny are closer to the female P. trichocarpa than male P. deltoides parent. Although the pure triploid progeny population did not have higher genetic variance in leaf traits than the pure diploid population, the former appears to hide much non-additive genetic variance and display strong genetic control over the phenotypic plasticity of leaf traits. It is suggested that the cryptic non-additive variance, especially epistasis, can be released when a population is disturbed by changes in the environment. A mixed diploid and triploid progeny population combines phenotypic and genetic characteristics of both pure hybrids and is considered to be of adaptive significance for populars to survive and evolve in a fluctuating environment. The significant effect due to general and specific combining ability differences at the population level suggests that the population divergence of these two species is under additive and non-additive genetic control. PMID- 10816979 TI - Mapping quantitative trait loci for principal components of bone measurements and osteochondrosis scores in a wild boar x large white intercross. AB - Data on osteochondrosis and femur dimensions from 195 F2 pigs from a wild boar x Large White intercross were analysed with the aim of detecting quantitative trait loci (QTLs) for normal and disturbed bone formation. The information from numerous recorded traits was summarized by principal component analysis and analysed by least-squares interval mapping. An increase in the proportion of wild boar alleles across the genome increased length versus width of femur and reduced the prevalence of osteochondrosis. The presence of QTLs with an impact on femur dimensions was indicated on chromosomes 2, 4, 16 and 17 and on osteochondrosis on chromosomes 5, 13 and 15. A substantial effect of the chromosome 5 QTL calls for further studies within commercial populations to evaluate whether marker-assisted selection could be used to reduce the prevalence of osteochondrosis. PMID- 10816980 TI - Markov chain Monte Carlo for mapping a quantitative trait locus in outbred populations. AB - A Bayesian approach is presented for mapping a quantitative trait locus (QTL) using the 'Fernando and Grossman' multivariate Normal approximation to QTL inheritance. For this model, a Bayesian implementation that includes QTL position is problematic because standard Markov chain Monte Carlo (MCMC) algorithms do not mix, i.e. the QTL position gets stuck in one marker interval. This is because of the dependence of the covariance structure for the QTL effects on the adjacent markers and may be typical of the 'Fernando and Grossman' model. A relatively new MCMC technique, simulated tempering, allows mixing and so makes possible inferences about QTL position based on marginal posterior probabilities. The model was implemented for estimating variance ratios and QTL position using a continuous grid of allowed positions and was applied to simulated data of a standard granddaughter design. The results showed a smooth mixing of QTL position after implementation of the simulated tempering sampler. In this implementation, map distance between QTL and its flanking markers was artificially stretched to reduce the dependence of markers and covariance. The method generalizes easily to more complicated applications and can ultimately contribute to QTL mapping in complex, heterogeneous, human, animal or plant populations. PMID- 10816981 TI - More about quantitative trait locus mapping with diallel designs. AB - We present a general regression-based method for mapping quantitative trait loci (QTL) by combining different populations derived from diallel designs. The model expresses, at any map position, the phenotypic value of each individual as a function of the specific-mean of the population to which the individual belongs, the additive and dominance effects of the alleles carried by the parents of that population and the probabilities of QTL genotypes conditional on those of neighbouring markers. Standard linear model procedures (ordinary or iteratively reweighted least-squares) are used for estimation and test of the parameters. PMID- 10816983 TI - Federal government issues "fraud alert" to physicians. PMID- 10816982 TI - Marker-assisted selection using ridge regression. AB - In cross between inbred lines, linear regression can be used to estimate the correlation of markers with a trait of interest; these marker effects then allow marker assisted selection (MAS) for quantitative traits. Usually a subset of markers to include in the model must be selected: no completely satisfactory method of doing this exists. We show that replacing this selection of markers by ridge regression can improve the mean response to selection and reduce the variability of selection response. PMID- 10816984 TI - Minimally invasive coronary artery bypass surgery. PMID- 10816985 TI - Hemophagocytic syndrome following an Epstein-Barr virus infection: a case report and literature review. AB - Hemophagocytosis is an uncommon disorder characterized by proliferation of histiocytes that actively engulf other hematopoietic cells causing cytopenia. Reactive or secondary hemophagocytosis is very rare in healthy adults in the US. Various infectious, as well as neoplastic and immunologic etiologies of reactive hemophagocytosis have been reported. It is a non-malignant, reactive disorder characterized by hemophagocytosis in the bone marrow and reticuloendothelial system (RES) resulting in pancytopenia, fever, hepatic dysfunction, and disseminated intravascular coagulation (DIC). No consensus exists in the literature regarding optimal treatment of virus-associated hemophagocytic syndrome (VAHS). We report a case of VAHS in a previously healthy immunocompetent male and review the diagnosis and management of this rare disorder. PMID- 10816986 TI - The burden of cancer in Kentucky: the 1997 Kentucky cancer incidence report. AB - The purpose of collecting this important disease burden information is of course not simply for reports like this one. The real value lies in use of data for program planning, resource allocation, program design, and evaluation. Data in the annual KCR report and on the website should be a valuable resource for district and county health departments in their assessment of health problems in their area and as critical decisions are made about how to utilize limited intervention resources. The Kentucky Cancer Registry will continue to work very closely with the Community Outreach Division of the Kentucky Cancer Program to make this data available on an annual basis to District Cancer Councils to help guide their assessment and planning process. District and local health departments are encouraged to actively participate in the District Cancer Councils and in the cancer control activities guided by the regional community outreach coordinators. PMID- 10816987 TI - Peril? Litigation! PMID- 10816988 TI - Molecular analysis of Duffy, Yt and Colton blood groups in Taiwanese, Filipinos and Thais. AB - We used a polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) method for DNA-based typing of Duffy, Yt and Colton blood groups in Taiwanese, Filipinos and Thais. A total of 200 Taiwanese, 115 Filipinos and 105 Thais were studied. In the Duffy blood group in Taiwanese, 180 cases (90%) were homozygote of Fya, 18 cases (9%) were double heterozygote of Fya and Fyb, and 2 cases (1%) were homozygote of Fyb. In Filipinos, 98 cases (85.2%) were homozygote of Fya, 16 cases (14.0%) were double heterozygote of Fya and Fyb and 1 case (0.8%) was homozygote of Fyb. In Thais, 87 cases (82.9%) were homozygote of Fya, 18 cases (17.1%) were double heterozygote of Fya and Fyb, and no case of Fyb was found. These results correlate well with serological phenotype. For the Yt blood group, only YT1 was found in Taiwanese. Among Filipinos, 114/115 (99.1%) was YT1/1 and 1/115 (0.9%) was YT1/2. In Thais, 103/105 (98.1%) was YT1/1 and 2/105 (1.9%) was YT1/2. For the Colton blood group, the results showed that there was only Coa allele in these three populations. Our results provide the first data of the Yt and Colton blood groups in these three populations. PMID- 10816989 TI - Hypothyroid myopathy-pathological and ultrastructural study. AB - Seventeen patients with hypothyroid myopathy were studied before and after thyroxine (T4) treatment. The severity of clinical myopathy was assessed with the aid of a modified rating scale. Laboratory findings including thyroid function, serum creatine kinase (CK), and electromyography were assessed at regular time intervals until a final muscle biopsy was performed. The average period of follow up was 1.8 years. The authors emphasize: 1) in skeletal muscle pathology of hypothyroidism, the fiber atrophy and increased central nuclear counts are evidence of clinical myopathy during thyroxine treatment; 2) in ultrastructural pathology, the abnormal glycogen accumulation accounts largely for clinical severity and its ongoing resolution is parallel to steady amelioration following T4 therapy, while mitochondrial abnormalities are insignificant in clinical correlation and probably become permanent in some cases with prolonged hypothyroidism; and 3) serial needle biopsies of skeletal muscle are impractical for long-term study of hypothyroid myopathy, but they may be reserved for those patients with a sustained myopathic complaints on T4 therapy. PMID- 10816990 TI - Core decompression for osteonecrosis of the femoral head at pre-collapse stage. AB - Twenty-five patients (27 hips) were retrospectively studied for core decompression in the treatment of osteonecrosis of the femoral heads at pre collapse stage, Ficat and Alert stages I and II, from Apr. 1984 to Jun. 1998 with follow-up period at least 1 yr (mean 28 months). Eleven hips (10 patients) were considered to have failed due to progressive collapsed more than 2 mm, severe pain or reoperation. We further analyzed the results with regard to the stages, the size of the necrotic area according to the Ohzono classification, the lateral head index (LHI) and the Kerboul combined necrotic angle. The survival rate according to the Ohzono classification was type 1B: 100%, 1C: 44%, 2: 25%, and 3B: 0%. Hips with Kerboul angle less than 250 degrees revealed satisfactory results in all except one hip. All hips with Kerboul angle more than 250 degrees collapsed. Fourteen of the 15 hips (93%) with lateral head index more than 20% did not collapse and all hips except one (7/8) with LHI less than 12% collapsed. We conclude core decompression yields satisfactory results in osteonecrotic femoral heads at pre-collapse stage and with small necrotic area or good lateral buttress. PMID- 10816991 TI - Strength and functional performance of patients with limb-girdle muscular dystrophy. AB - This study was design to reach a better understanding of muscle strength, motor function and activity of daily living in patients of limb-girdle muscular dystrophy. Forty-eight patients diagnosed as cases of limb-girdle muscular dystrophy were included in this study. Manual muscle testing was used to evaluate muscle strength. The Brooke and Vignos scales were used to grade upper and lower extremities function, respectively, and the ability of daily living activity was measured by Barthel index. Our patients showed progressive symmetrical limb girdle muscular weakness. Upon regression analysis we found that mean muscle strength was inversely related to disease duration (years) as follows: mean muscle strength = 0.6052 + (0.6309/disease duration). According to the Brooke functional scale, 89.6% of our patients were graded as 1-3 and 10.4% were graded as 5. On the Vignos functional scale, 79.1% of patients fell into the grades 1-5, one person (2.1%) in grade 6 and 18.8% in grade 9 category. The average Barthel index was 85.3 +/- 20.7. Mean muscle strength was significantly correlated with the average Barthel, Vignos and Brooke functional scales. Our study could offer the strength and functional performance of limb-girdle muscular dystrophy on natural history. The muscle strength declined in Taiwanese patients of limb girdle muscular dystrophy in a typical pattern. Regression analysis showed that the strength was inversely related to disease duration. These findings demonstrate that most of our patients suffered from mild or moderate physical disability. PMID- 10816992 TI - Intravesical mitoxantrone in superficial bladder cancer. AB - Twenty-five patients with Ta-T1 transitional cell carcinoma of the bladder were treated by transurethral resection (TUR) followed by intravesical chemoprophylaxis with mitoxantrone (10 mg diluted in 50 ml normal saline) administered weekly for 6 weeks. After a mean follow-up of 12 months, 76% of patients in the whole group, 69% in the newly diagnosed patients and 89% in the group of previously relapsed patients remained relapse free. These rates compare favorably with the other prophylactic agents available. Therapy was well tolerated in most patients, with only two patients reporting grade 3 local toxicity. We conclude that mitoxantrone is an effective and safe agent for intravesical chemotherapy. PMID- 10816993 TI - Ptosis surgery on chronic myasthenia gravis. AB - We report three cases of long-standing, early onset myasthenia gravis. The eyes of all three patients eventually became disabled, and the patients were almost unable to move their eyes in any direction. We termed this condition as "frozen eyes" appearance. We discuss the unique clinical condition and follow the electromyographic and neuro-ophthalmological characteristics. The patients all suffered from ptosis, mild to moderate exotropia, and facial and oropharyngeal weakness. All of the patients responded to the neostigmine test positively at the early stage. Two of them were seropositive. The results of electromyography for two of these three cases were abnormal. The average duration of follow-up was 5 years. The treatments included conventional recession and resection procedures with adjustable sutures for strabismus and frontalis suspension for blepharoptosis. The patients achieved stable satisfactory results in over one-and a-half years of postoperative follow up. PMID- 10816994 TI - Corneal subepithelial opacities associated with inappropriate contact lens care. AB - PURPOSE: To report two cases of corneal subepithelial opacities, which may be related to improper lens care by the patients. METHODS: Slit-lamp examination and monitoring of contact lens care procedures. RESULTS: Two female patients experienced similar symptoms of soreness, blurring, and red eyes after wearing soft contact lenses. Slit-lamp examinations demonstrated numerous subepithelial opacities in the central cornea. Both patients used the same kind of cleaner (MiraFlow) inappropriately, which resulted in the possibility of residual cleaner remaining in the lenses. The symptoms and signs gradually subsided after discontinued use of soft contact lens and application of topical steroid. CONCLUSIONS: Residual MiraFlow in soft contact lenses may result in corneal subepithelial opacities. PMID- 10816995 TI - Major hemorrhage in a patient with multiple submucous leiomyomata during the treatment of long-acting gonadotropin-releasing hormone agonist. AB - Gonadotropin releasing hormone agonist (GnRH-agonist) therapy has been useful as an adjunct before myomectomy or hysterectomy for uterine myoma but the concealed risk is often overlooked. We report an extremely rare clinical presentation of a patient with multiple submucosal myomata during the treatment of long-acting gonadotropin-releasing hormone agonist (GnRH-agonist) in a 23-year-old, virgin woman. This patient exhibited heavy menstruation and severe anemia for half of a year. Ultrasound demonstrated multiple submucous myomata and intramural myomata. She received a conservative medical treatment by GnRH-agonist. The patient showed marked suppression of serum estradiol concentrations throughout treatment (< 20 pg/ml since first dose injection). The volume of the uterus decreased 21% and the total volume of the uterine myomata decreased 27% at the end of the second dose injection. However, a sudden onset of major hemorrhage occurred at the 65th day without "add-back" hormonal replacement therapy after initial therapy of GnRH agonist. Hypovolemic shock followed soon and immediately resuscitation was performed. After resuscitation, the patient was treated with hysteroscopic myomectomy, followed by 30 ml balloon Foley catheter placement for compressing the intrauterine rough surface and hormonal replacement therapy. When uterus returned to the normal size at the end of the first week, intrauterine device was positioned and maintained for three months. The patient married four months later and got pregnant soon. Now she has a pregnancy of 22 gestational weeks. The phenomenon suggests presence of concealed and potential risk of GnRH-agonist for managing a patient with multiple submucous myomata, even though GnRH-agonist is a well-documented transient treatment for uterine myomata not only by its effect on tumor shrinkage and decreasing blood loss during the myomectomy but also by providing a time for hematological recovery. This unexpected and unwanted clinical presentation should be alerted. PMID- 10816996 TI - [Recommendations for the perioperative management of heart disease patients in non-cardiac surgery. Societa Italiana de Anestesia, Analgesia, Rianimaione e Terapia Intensiva]. PMID- 10816997 TI - [When does accidental hypothermia become voluntary hypothermia?]. PMID- 10816998 TI - [In Process Citation] PMID- 10816999 TI - Critical point in sepsis: VO2/DO2 controversy. PMID- 10817000 TI - Effective doses of cisatracurium. Potentiation by sevoflurane and increasing requirements with age. AB - BACKGROUND: The aim of the study was to investigate whether or not sevoflurane at different concentrations influenced the ED(effective dose) 50, 90 and 95 of cisatracurium and whether there is any relationship with age. METHODS: Individual dose response curves for cisatracurium were constructed in 80 consecutive patients randomly assigned to four groups: group 1 TIVA with fentanyl and propofol; group 2, 3 and 4 sevoflurane 1, 1.5, 2 MAC respectively, MAC age adjusted and with N2O 65%. Patients were premedicated with diazepam and induction accomplished with fentanyl (1.5-2 micrograms/kg) and propofol (1.5-2 mg/kg); following intubation with succinylcholine (0.4 mg/kg), TIVA or sevoflurane at the desired MAC was maintained until equilibration occurred and then cisatracurium administered in 8-10 micrograms/kg boluses until the attainment of ED95. Neuromuscular monitoring utilized TOF stimulation and ratio evaluation between Ist twitch of the train and basal twitch (T1/Tc) and continuous recording of accelerometry (Tof-guard). Single dose response curves were analyzed with sigmoid equations; the Eds and other parameter means were subjected to analysis of variance for parametric and non parametric data. RESULTS: More boluses were required under TIVA and sevo 1 MAC anesthesia (8.3 and 7.1 vs 4.9 of sevo 1.5 and 2 MAC); maximal depression (94.95%) was reached more frequently with sevo, while under TIVA an average 90% depression only was obtained. Mean ED50 (microgram/kg) where 22 (sevo) vs 34 (TIVA): ED90 averaged 36 (sevo) vs 53 (TIVA); ED95 averaged 39 (sevo) vs 56 (TIVA), with significant differences (p < 0.05) between Eds obtained from TIVA group in comparison with sevoflurane groups, but without differences between averaged EDs obtained with different MAC values among sevoflurane groups. The investigation of the relationship between age and Eds demonstrated a linear regression for all groups considered together (p = 0.000) for all Eds (50, 90, 95). CONCLUSIONS: In conclusion, the presence of MAC multiples of sevoflurane potentiated only slightly the dosages of cisatracurium and EDs linearly increase with age, particularly under sevoflurane anesthesia. PMID- 10817001 TI - [The use of alfentanil for short duration surgery in pediatric anesthesia]. AB - BACKGROUND: In the present paper, the effectiveness of the alfentanyl-propofol combination versus fentanyl-propofol association in pediatric anesthesia during short surgical procedures was investigated. METHODS: EXPERIMENTAL DESIGN: randomized study. SETTING: This study was carried out at the Surgical-Division of the Pediatric Clinic of the University "La Sapienza", Rome. PATIENT: 100 children (age 4.95 +/- 0.26 yr, body weight 21.13 +/- 0.86 kg) undergone to short anesthesia (50 +/- 0.01 min), were examined. PATIENTs were divided randomly into two groups, A and B, each composed of 50 children. PATIENTs in group A received an alfentanyl bolus dose of 50 micrograms.kg-1 and a continuous infusion of 0.50 microgram.kg-1.min-1, whereas patients in group B received a fentanyl bolus dose of 5 micrograms.kg-1 and a continuous infusion of 0.16 microgram.kg-1.min-1. In all patients, propofol was given in the same dose (bolus dose 1 mg.kg-1; continuous infusion 0.1 mg.kg-1.min-1) in association with opioid analgesic. MEASUREMENTS: non-invasive arterial pressure, EKG, heart rate, rectal temperature, oxygen saturation, capnography; during the postoperative course, we evaluated the motor activity response, the degree of weakfulness and consciousness. RESULTS: No significant differences between two groups in the systolic, diastolic and mean arterial pressure were pointed out whereas a decrease in heart rate in the group B and a better and more rapid restoration of breathing in group A was observed. Children of group A, after awakening, were more quiet and answered more readily to simple orders (64%). In group B 50% of children showed psychomotor unrest and 10% of them cried. The group that received alfentanyl answered more readily than group B (42% versus 26%). CONCLUSIONS: The present study shows that alfentanyl allows a better control of the surgical analgesia and it assures a good analgesic cover for all short surgical procedures without risk of early respiratory depression in postoperative period. Furthermore alfentanyl assures an early postoperative recovery due to the minimal pool, minimal undesired effects, and a cardiocirculatory stability. PMID- 10817002 TI - Unilateral spinal anesthesia or combined sciatic-femoral nerve block for day-case knee arthroscopy. A prospective, randomized comparison. AB - BACKGROUND: To evaluate the time required to achieve surgical block and fulfill standardized discharge criteria in outpatients receiving knee arthroscopy with either unilateral spinal anesthesia or combined sciatic-femoral nerve block. METHODS: After a standard midazolam/ketoprofen premedication and baseline measurement of cardiovascular parameters, 50 ASA physical status I-II patients scheduled for elective outpatient knee arthroscopy were randomized to receive unilateral spinal anesthesia with 8 mg of 0.5% hyperbaric bupivacaine injected without barbotage through a 25-gauge Whitacre spinal needle (group USA, n = 25), or combined sciatic-femoral nerve block with 25 ml of 2% mepivacaine (15 ml for femoral nerve block and 10 ml for sciatic nerve block) (group SFNB, n = 25). Times from local anesthetic injection to achievement of surgical block defined as the presence of adequate motor (complete motor blockade of the operated limb in the USA group and inability to move the ankle and the knee of the operated limb in the SFNB group) and sensory (loss of pinprick sensation at T12 on the operated side in the USA group, or in the femoral and sciatic nerves distribution in the SFNB group) blocks was recorded. Times to block resolution, urination, unassisted ambulation, and readiness to home discharge were also recorded by a blind observer, as well as occurrence of untoward events during surgery. RESULTS: Surgical block was achieved in 15 +/- 6 min in group USA and 16 +/- 6 min in group SFNB (p = NS). No differences in hemodynamic undesired effects and success rate were observed. Even though USA patients showed a faster time to ambulation (166 +/- 44 min versus 217 +/- 49 min, p = 0.002) and later urination (240 +/- 90 min versus 145 +/- 36 min, p = 0.0001) than SFNB group, no differences in home discharging were reported (246 +/- 98 min versus 211 +/- 77 min, respectively). Bladder catheterization was required in 2 patients of USA group only; however, no patient was admitted to the hospital because of urinary retention. CONCLUSIONS: In outpatient knee arthroscopy, a combined sciatic-femoral nerve block with 2% mepivacaine provides similarly successful anesthesia with onset times and home discharge similar to those provided by unilateral spinal anesthesia. PMID- 10817003 TI - Customization of SAPS II for the assessment of severity in Italian ICU patients. ARCHIDIA. Archivio Diagnostico. AB - BACKGROUND: To assess the accuracy of SAPS II on the Italian population and to perform a customization of the model. DESIGN: observational prospective study. PATIENTS: 24 participating centers. 6794 patients out of 9185 enrolled in the cohort study. MEASUREMENTS: the performance of SAPS II was assessed with calibration and discrimination. In the case of standard model not fitting the data, new logistic regression equation has been calculated using Archidia database. RESULTS: SAPS II showed a good discrimination, but a bad calibration. The new logit has been calculated on the population examined in 1995 and successively tested on the population collected during 1996. In both samples discrimination and calibration results were good. CONCLUSIONS: Our customized model reaches a high standard in calibration and discrimination. We suggest this model for future application in Italian ICU. PMID- 10817004 TI - [Comparative study of the effects of morphine and tramadol in the treatment of postoperative pain]. AB - BACKGROUND: To compare morphine and tramadol in the treatment of postoperative pain (POP), using a system of Patient Controlled Analgesia (PCA). METHODS: EXPERIMENTAL DESIGN: Randomised comparative study on 88 patients, undergoing general major surgical operations (gastrectomy, colectomy and hemicolectomy). PATIENTS: PCA i.v.: group M treated with morphine (at a dilution of 1 mg/ml) and group T with tramadol (at a dilution of 10 mg/ml). Loading dose: 0.05 mg/kg for the group treated with morphine and 0.5 mg/kg for the group treated with tramadol. Baseline infusion: 1.5 ml/h. Bolus on demand: 0.2 ml every 30 minutes. At T0, after 1 hour, 3 and 18 hours assessment of pain intensity, level of sedation, respiratory and cardiocirculatory parameters. After 18 hours assessment of the amount of drug administered, the number of boluses requested and given and any side effects. STATISTICAL ANALYSIS: ANOVA test, Student "t", chi 2. RESULTS: A statistically significant reduction in POP was found after 1 hour, 3 and 18 hours with no significant differences between the two groups. No statistically significant differences in cardiocirculatory or respiratory parameters or level of sedation were found. Few side effects were observed. CONCLUSIONS: Tramadol determines pain relief similar to morphine, with the advantage that it does not lead to abuse, tolerance or addiction. PMID- 10817005 TI - [Injury of the long thoracic nerve after LRA, which etiology? Clinical case]. AB - The long thoracic nerve supplies the serratus magnus muscle. Its injury causes "winging" of the scapula and functional limitation of the upper limb. Such lesion is mostly due to chronic trauma, occasionally iatrogenic. This case report presents a case of serratus magnus muscle paralysis following an anesthetic block of the brachial plexus performed at the time of a minor orthopedic surgery of the wrist, in a young healthy woman who was a tennis player. The observed lesion seemed to have been caused by the anesthetic technique employed, but, at the same time, it is impossible to rule out, in the etiology of the disease, a role of other predisposing factors, such as the intense sport activity above mentioned, or of concurrent factors, such as an incorrect position in the immediate postoperative period. PMID- 10817006 TI - [The predictive lidocaine test in treatment of neuropathic pain]. AB - BACKGROUND: In daily practice, the intravenous lidocaine drip has been introduced as a predictive test for subsequent oral treatment with adjuvant drugs (anti depressants, channel blockers and anti-convulsants). Our aim is the assessment of the correlation between the test response and the effectiveness of the consequent oral drug therapy. METHODS: 183 inpatients (central and peripheral neuropathic pain), treated between 1996-1997, were retrospectively checked. The trial was conducted as follows: a VAS reading > 5 was taken; a subsequent continuous i.v. lidocaine drip was given, at a dose of 4 mg/kg, in saline solution; a VAS reading was taken before (VAS 0), every 5 minutes, and at the end of the drip (VAS 1); the results of the drip were to be considered positive where pain relief was > or = 50%; irrespective of test results, all patients were given a different follow up drug therapy; a VAS reading was taken one month after the drug therapy (VAS 2). RESULTS: Eighty-five patients (90%), responders to lidocaine, had a pain relief and 71 patients (85%), no responders to lidocaine, did not have improvement by taking oral drugs. CONCLUSIONS: In agreement with a other authors, we also noted that there was a statistically significant correlation between the results obtained and the therapy prescribed; the usefulness of a lidocaine drip as routine procedure to predict the therapeutic response of neuropathic pain to adjuvant analgesics is underlined. PMID- 10817007 TI - [General anesthesia and undiagnosed cerebral metastasis. Clinical case]. AB - A case of late recovery from anesthesia in a patient undergoing bronchoscopy for surgical removal of metastatic bronchial mass is presented. The patient was comatose and a CT scan revealed the presence of bleeding inside a tumor, probably a metastasis, located in the right cerebellum. This report demonstrates that undetected cerebral metastases might lead to late recovery from anesthesia and underlines that accurate neurologic examination is mandatory in patients affected by tumors potentially spreading to the brain. PMID- 10817008 TI - [Continuous updating of italian certification in Anesthesiology-resuscitation]. PMID- 10817009 TI - [Treatment of alcoholic liver diseases. Abstinence, nutritional support, drug therapy, liver transplantation]. AB - The review summarizes clinically established treatment forms of alcoholic liver disease in four main chapters: abstinence, nutritional supportation, drug therapy and liver transplantation are discussed. Drug therapy is described according to the three types of alcoholic hepatopathies (fatty liver, hepatitis and cirrhosis). Early diagnosis and treatment depending on the severity and stage of alcoholic liver disease are of importance for the attempts to retard progression and improve prognosis. PMID- 10817010 TI - [Combination of hormone replacement therapy with buspirone in the treatment of climacteric complaints]. AB - At the menopause outpatient unit among 100 patients with climacteric problems 50 were treated with continuous hormone replacement therapy alone (Group A), and in 50 other cases were given a combination of a half dose hormone replacement therapy (HRT) and anxiolytic drug (Group B). After a 3 month treatment period the complaints were reduced at the same extent in both groups. The frequency of the side effects was similar in both groups. The combination of a reduced dose of HRT and an anxiolytic drug can be an alternative treatment to patients refusing HRT because of cancerophobia or side effects and to those, whom the reduced dose of HRT might be sufficient to avoid climacteric complaints. PMID- 10817011 TI - [Comparative study of the attitudes of students and nurses to physician-assisted suicide--first results]. AB - While the popular media and the professional literature deal with the topic of euthanasia intensively, the problems of physician-assisted suicide received insufficient attention in Hungary. The authors review the most important details of the physician-assisted suicide. A twelve-item scale to measure attitude toward PAS (constructed and validated by G. Domino) was administered to the samples of Hungarian social science, medical students and nurses. The distributions of responses for the 12 items for the social and medical samples are compared and discussed. Also a cross-cultural comparison was made with an American student sample examined by Domino. The social science students who have the fewest personal experiences with serious, or terminally ill patients are the most liberal group, characterised by the most permissive attitudes toward PAS. Nurses who have everyday contact and experience with these patients are the most conservative; they show more or less conclusively the lowest acceptability rate of PAS. The attitudes of medical student's group, of the would-be physicians are between them, and they are the most controversial and ambivalent. Future research is necessary to get more information about attitudes toward physician-assisted suicide. PMID- 10817012 TI - [Minimally invasive adrenalectomy with posterior retroperitoneoscopy]. AB - Authors report a total of 14 adrenalectomies performed from a posterior (lumbotomy) approach, using minimally invasive retroperitoneoscopic technique. The "UltraCision" scalpel, an ultrasound activated cutter-coagulation device has been used for operative tissue dissection. Two cases were converted. The mean operating time of the successful 12 cases were 128 minutes. No mortality and no septic complication occurred. The mean operative blood loss of the 12 procedures were less than 100 ml. The mean hospitalisation was 3-5 days, and the complete recovery needed 2-3 weeks. On the basis of our own experiences and the ones of prospective, randomized clinical studies of the literature, the retroperitoneoscopic adrenalectomy from a posterior approach is recommended for the surgical treatment of benign tumours of the suprarenal gland with a size less than 5 cm diameter. The low conversion- and complication-rate, the minimal operative blood loss, the short hospitalization and quick recovery time all are the advantages of this method. It can be performed after previous abdominal operation and in cases with morbid obesity. The technique is suitable for bilateral adrenalectomies as well. This method is not justified for the removal of malignant and/or larger than 5 cm adrenal tumours. Coagulopathies are contraindications. PMID- 10817013 TI - [Dr. Pal Fromm, the first nursery physician in Hungary, military physician to Lajos Kossuth, medical officer]. PMID- 10817014 TI - ["They look after their health": matters of health in the habits and life of ancient Egyptians]. PMID- 10817015 TI - [About the treatment of allergic respiratory diseases (Histamine, histaminase, histamine-azoprotein, antihistamines). 1950]. PMID- 10817016 TI - The pedunculopontine nucleus: its role in the genesis of movement disorders. AB - The pedunculopontine nucleus (PPN) is located in the dorso-lateral part of the ponto-mesencephalic tegmentum. The PPN is composed of two groups of neurons: one containing acetylcholine, and the other containing non-cholinergic neurotransmitters (GABA, glutamate). The PPN is connected reciprocally with the limbic system, the basal ganglia nuclei (globus pallidus, substantia nigra, subthalamic nucleus), and the brainstem reticular formation. The caudally directed corticolimbic-ventral striatal-ventral pallidal-PPN-pontomedullary reticular nuclei-spinal cord pathway seems to be involved in the initiation, acceleration, deceleration, and termination of locomotion. This pathway is under the control of the deep cerebellar and basal ganglia nuclei at the level of the PPN, particularly via potent inputs from the medial globus pallidus, substantia nigra pars reticulata and subthalamic nucleus. The PPN sends profuse ascending cholinergic efferent fibers to almost all the thalamic nuclei, to mediate phasic events in rapid-eye-movement sleep. Experimental evidence suggests that the PPN, along with other brain stem nuclei, is also involved in anti-nociception and startle reactions. In idiopathic Parkinson's disease (IPD) and parkinson plus syndrome, overactive pallidal and nigral inhibitory inputs to the PPN may cause sequential occurrences of PPN hypofunction, decreased excitatory PPN input to the substantia nigra, and aggravation of striatal dopamine deficiency. In addition, neuronal loss in the PPN itself may cause dopamine-resistant parkinsonian deficits, including gait disorders, postural instability and sleep disturbances. In patients with IPD, such deficits may improve after posteroventral pallidotomy, but not after thalamotomy. One of the possible explanations for such differences is that dopamine-resistant parkinsonian deficits are mediated to the PPN by the descending pallido-PPN inhibitory fibers, which leave the pallido-thalamic pathways before they reach the thalamic targets. PMID- 10817017 TI - The effective concentration and exposure time of mitomycin-C for the inhibition of lens epithelial cell proliferation in rabbit eyes. AB - The proliferation of residual lens epithelial cells following cataract surgery is assumed to be a major cause of posterior capsular opacification. To assess the efficacy of mitomycin-C in preventing posterior capsular opacification, we determined the effective concentration and exposure time of mitomycin-C in inhibiting rabbit lens epithelial cell proliferation. The fourth-passaged rabbit lens epithelial cells were maintained for one day and then exposed to mitomycin-C for 1, 2, 3, and 5 minutes, respectively. There were 9 different plating concentrations of mitomycin-C with two-fold serial dilution. The maintenance of the phenotypic properties of lens epithelial cells was confirmed by continuous transcription of lambda-crystalline mRNA determined by reverse transcription polymerase chain reaction and the polymorphism of the restriction fragment. Cell proliferation was assayed with 3H-thymidine incorporation into DNA. The fourth passaged cells maintained the expression of lambda-crystalline mRNA, suggesting that they are phenotypically authentic lens epithelial cells. The effective concentrations and exposure time of mitomycin-C were 0.1 mg/ml for 1 minute and 2 minutes, and 0.025 mg/ml for 2 minutes. By these results, we postulated that mitomycin-C at relatively short incubation times could be clinically used for prevention of posterior capsular opacification after cataract surgery. PMID- 10817018 TI - A clinical analysis of 11 patients with chronic actinic dermatitis in Korea. AB - Chronic actinic dermatitis is a rare disease worldwide and also in Korea. However there has been no clinicohistologic and photobiological analysis of chronic actinic dermatitis in Korea. We examined 11 patients who were diagnosed as chronic actinic dermatitis and the results of this study were compared with previous reports. Most patients were elderly men who had erythematous papules or lichenified plaques on sun-exposed areas such as the face, neck, and dorsum of the hands with severe itching sensation. All patients had severe sensitivity to UVB and biopsied specimens showed findings of chronic eczema. Five patients had positive photopatch test materials. The patients were treated with systemic and topical steroid, cyclosporine and antihistamine. PMID- 10817019 TI - Early ultrastructural changes of apoptosis induced by fumonisin B1 in rat liver. AB - A time sequential study was performed to investigate the histological and ultrastructural findings of fumonisin B1-induced apoptosis in the male Sprague Dawley rat liver. Six hours after administration of FB1, marked morphologic changes of hepatocytes included the appearance of small vacuoles along the margin of cell membrane. Twelve hours after injection of FB1, acidophilic degeneration of cells occurred, but no fragmented nucleus was evident around the centrilobular area, with few apoptotic cells. By electron microscope, the degenerated acidophilic cells revealed following changes: characteristic formation of cytoplasmic vacuoles, condensed cytoplasm, detachment from neighboring cells, and as well as margination of nuclear chromatin and swollen mitochondria with amorphous matrical deposit. The number of apoptotic cells or bodies was further enhanced at 24 hours in the vicinity of dense acidophilic cells, resulting in a marked increase over the values of control rats. Serum analysis revealed the elevation of cholesterol levels from the beginning to the end of this experiment. Morphologic data and serum findings in this study support the theory that FB1 induced alteration of membrane lipid constituents of the hepatocytes are likely to be early key events in explaining the FB1 apoptotic effect. PMID- 10817020 TI - Masturbation and its relationship to sexual activities of young males in Korean military service. AB - This study examined the masturbatory experiences and other sexual activities of young Korean males in military service. The actual status of masturbation and its relationship to sexual activity questionnaire. A total of 1,212 young males among military personnel in Korea were interviewed with sexuality questionnaires on masturbation, sexual intercourse and personal characteristics. We divided these subjects into four groups according to the age of initiation of masturbation and analyzed the relationship between masturbation and other sexual activities. The mean age of subjects was 22.03 +/- 1.22 (19-27) years. The percentage of men who reported ever having masturbated was 98.1% (1189/1212) and the average age of initiation of masturbation was 14.26 +/- 1.66 years. So we divided 1,212 males into four groups on the basis of the average age below and above one standard deviation. Overall, 67.7% (821/1212) had experienced sexual intercourse, and significantly, the earlier the initiation age of masturbation, the higher the coitus rate in each group (p < 0.05). About 21.5% of the men who were not virgins had experienced their first sexual intercourse with prostitutes. The mean age of first coitus, the incidence of sexually transmitted disease (STD) and the frequency of masturbation were closely linked to the initiation age of masturbation, respectively (p < 0.05). A masturbatory guilt feeling was seen in about 10.9% (132/1212) and there was no significant difference according to the types of religious worship (p = 0.227). On the basis of this study, sexual activities generally increased accordingly as the beginning of masturbation was earlier. Coital incidence in this study was 67.7% for young males in Korean military service, and 21.5% of them had their first sexual intercourse with prostitutes. Prostitution still plays an important role in the sexual lives of males in Korea. The incidence of STD was over 10% and homosexual manifestation was seen in 1.07% of subjects. Thus a proper and sound sex education at school during childhood is needed in Korea. A large scale survey of sexual behavior in various populations is essential for the improvement of correct sexual concepts. PMID- 10817021 TI - The classification of segmental vitiligo on the face. AB - Segmental vitiligo usually has an onset early in life and spreads rapidly within the affected area. Among 1,300 patients with vitiligo, 191 patients with segmental vitiligo involving the face were evaluated. In this study, the distribution of segmental vitiligo on the face could be classified into 5 patterns which have distinctive features. This classification of facial segmental vitiligo can provide some indication of the future distribution of early lesions if they have begun to spread. PMID- 10817022 TI - Endoscopic snare resection for tumors of the ampulla of Vater. AB - Though surgical resection has been the traditional treatment for tumors of the ampulla of Vater, endoscopic maneuvers such as snare resection, laser photodestruction and electrofulguration have recently been introduced to avoid operation-related morbidity and mortality. From 1994 to 1996, 6 patients with ampullary tumor were managed by endoscopic snare resection and regularly followed. Endoscopic snare resection of the ampullary tumor was technically feasible in all patients and each procedure was performed in a single session. Histologic diagnoses of the resected specimens were adenoma in 4 patients and adenoma with coexistent adenocarcinoma in 2 patients. Resection margins were negative in all patients except 1 with coexistent adenocarcinoma and a radical pancreaticoduodenectomy was performed in that case. For the other patient with adenocarcinoma foci, no further treatment was persued since he was 72-year-old and refused operation. Acute pancreatitis developed in 2 patients after endoscopic therapy, but was resolved with conservative management. There was no procedure-related death. Surveillance duodenoscopy performed at 1 and 6 months after endoscopic resection revealed no evidence of recurrent tumor in 4 patients with adenoma. Among them, 3 patients are alive without evidence of recurrence at 16-37 months after resection, but 1 patient was lost after 9 months of follow-up. The patient with adenocarcinoma in whom a pancreaticoduodenectomy was performed, has been alive without recurrence for 12 months. Oral 5-fluorouracil was administered for the other patient with adenocarcinoma foci. Though he experienced local recurrence at 13 months after the procedure, he has been alive for 28 months after resection. In conclusion, endoscopic snare resection may be applied as a viable alternative to surgery in selected patients with small ampullary tumors. PMID- 10817023 TI - The prevalence and associated risk factors of renal artery stenosis in patients undergoing cardiac catheterization. AB - Renal artery stenosis may be a cause of hypertension and a potential contributor to progressive renal insufficiency. However, the prevalence of renal artery disease in a general population is poorly defined. The purposes of this study were to evaluate the prevalence of angiographically-determined renal artery narrowing in a patient population undergoing routine cardiac catheterization, and to identify the risk factors for renal artery stenosis. After left ventriculography, abdominal aortography was performed to screen for the presence of renal artery stenosis. A total of 427 patients (274 males, 153 females) were studied and the mean age was 59 years. Renal artery narrowing was identified in 10.5% of patients. Significant (> or = 50% diameter narrowing) renal artery stenosis was found in 24 patients (5.6%) and insignificant stenosis was found in 21 patients (4.9%). Significant unilateral stenosis was present in 4.2% of patients and bilateral stenosis was present in 1.4%. The stem of the renal artery was a more common site of stenosis in 62.2% of patients than in the ostium (37.8%), but the severity of stenosis was not significantly different according to the site of stenosis. By univariate and multivariate logistic regression analysis, the association of clinical variables with renal artery stenosis was assessed. Multivariable predictors included age, hypertension and peripheral vascular disease (p < 0.05). The variables such as sex, smoking history, hyperlipidemia, renal insufficiency, as well as the presence of obesity, severity of coronary heart disease and D.M., were not associated. In conclusion, the prevalence of angiographically-determined renal artery narrowing in a patient population undergoing cardiac catheterization is 10.5%. Old age, hypertension and evidence of peripheral vascular disease represent the predictors of renal artery stenosis. PMID- 10817024 TI - The prediction of adverse pregnancy outcome using low unconjugated estriol in the second trimester of pregnancy without risk of Down's syndrome. AB - To investigate the relationship between low unconjugated estriol (uE3) levels in the second trimester and adverse perinatal outcomes in pregnancies without increased risk for Down's syndrome, 1,096 women under 35 years of age underwent a mid-trimester AFP-hCG-uE3 screening test between January 1995 and June 1998. Multiple pregnancies, maternal diabetes, smoking and elevation of AFP and hCG levels more than 2.0 multiple of median (MoM) were excluded from our study population. The results were divided into a low-uE3 group with uE3 levels at or below 0.75 MoM and a normal uE3 group with uE3 levels above 0.75 MoM. The risk for adverse pregnancy outcome was compared between the two groups and the role of low uE3 as a predictor of adverse pregnancy outcome was determined. The data were assessed using chi 2 or Fisher exact test and then logistic regression was used for the final analysis. The odds ratio (OR) and corresponding 95% confidence intervals (CI) were also calculated. Unconjugated E3 levels at or below 0.75 MoM was significantly associated with fetal growth restriction after adjustment for maternal age, weight, sampling weeks, AFP and hCG levels (OR 0.413, 95% CI 0.174 0.900; P = 0.035). Low uE3 levels in the second-trimester could help in the detection of fetal growth restriction by a low risk group in Down's syndrome. Careful gestational dating and serial clinical and sonographic assessment of fetal growth may be required for the clinician to manage these parturients. PMID- 10817025 TI - Usefulness of a developed four-channel EGG system with running spectrum analysis. AB - Electrogastrography (EGG) is a method of measuring action potentials on the abdomen. It is noninvasive, inexpensive and easy to measure. However, the EGG signal has a very low frequency (0.05 Hz) and an extremely low amplitude (10-100 microV). Consequently, its measured waveform is difficult to analyze and it is not yet completely understood. In this study, a four-channel EGG measurement system was built to measure the action potential of the stomach. This system was compared with the commercially available one-channel Digitrapper EGG. The 3 cpm percentages were compared between the best channel of the four-channel system and channel 1, whose electrode position was similar to the commercially available one channel system for normal subjects. The average 3 cpm percentage of the best channel and channel 1 for preprandial measurement was 89.5% and 83.2%, respectively, and this was statistically significant (p < 0.001). Also the average 3 cpm percentage of the best channel and channel 1 for postprandial measurement was 90.4% and 76.5%, respectively, and this was statistically significant (p = 0.003). From these results, it can be concluded that a multi channel EGG system is required for better EGG measurement. PMID- 10817026 TI - The effect of pelvic floor muscle exercises on genuine stress incontinence among Korean women--focusing on its effects on the quality of life. AB - This study's purpose was to compare the treatment efficacy and the effects on the patients' quality of life of the pelvic floor muscle (PFM) exercise and the functional electrical stimulation (FES)-biofeedback method. Ninety female incontinence patients were randomly selected and evenly divided into three groups: control, intensive PFM exercise, and FES-biofeedback groups. They were treated for 6 weeks. The subjective changes in the severity of incontinence and discomfort in daily and social life were measured using a translated version of Jackson's Bristol female urinary symptom questionnaire. Objective changes of pelvic muscle contraction force were measured by perineometer. Pre and post treatment maximal pelvic floor muscle contractile pressure (PMC pressure) among the three groups showed statistically significant differences (p < 0.001). Especially the FES-biofeedback group showed significantly increased maximal PMC pressure compared with other groups (p < 0.001). From the questionnaire, pre and post-treatment changes in the severity of urinary incontinence and discomfort due to incontinence showed significant differences among the three groups (p < 0.001). The level of discomfort in daily life, social activity, physical activity, personal relations and discomfort due to urinary symptoms had largely changed and the FES-biofeedback group, in particular, showed a significant decrease after treatment. In conclusion, when PFM exercise and FES-biofeedback were compared in terms of their effects on the patients' quality of life, FES biofeedback proved to be more effective than verbal explanation or simple PFM exercise. PMID- 10817027 TI - Adrenergic sensitivity of uninjured C-fiber nociceptors in neuropathic rats. AB - We investigated the adrenergic sensitivity of afferent fibers in the L4 dorsal roots of rats with a unilateral ligation of the L5-L6 spinal nerves. About 12% of nociceptive fibers on the affected side were excited by sympathetic stimulation or by intra-arterial injection of norepinephrine which did not affect A beta fiber activity. Sympathetic excitation of nociceptive fibers was suppressed by alpha 1-antagonist prazosin, while it was unaffected by alpha 2-antagonist yohimbine. Most of these fibers were excited by intra-arterial injection of alpha 1-agonist phenylephrine, without being affected by an injection of alpha 2 agonist clonidine. Sympathetic excitation was blocked by lidocaine applied near the receptive fields of recorded fibers. The results suggested that some nociceptors remaining intact after partial nerve injury become sensitive to sympathetic activity by the mediation of alpha 1-adrenoceptors in the peripheral endings. PMID- 10817028 TI - Prenatal genetic diagnosis from maternal blood: simultaneous immunophenotyping and FISH of fetal nucleated erythrocytes isolated by negative and positive magnetic activated cell sorting. AB - Fetal nucleated red blood cells (nRBCs) are rare in maternal circulation, but their presence constitutes a potential source of non-invasive prenatal genetic diagnosis. This study was undertaken to establish a non-invasive prenatal genetic diagnosis method using isolated fetal nRBCs. A multi-step method including triple density gradient and magnetic activated cell sorting (MACS) using CD45 and CD71, cytospin centrifugation, K-B staining, and glycophorin A-immuno fluorescence in situ hybridization (GPA-immuno FISH) was performed. The study population included 65 patients from 8 to 41 weeks of gestation, and fetal nRBC was separated from all cases. The number of fetal nRBCs retrieved was 12.8 +/- 2.7 in 8 to 11 gestational weeks, 15.2 +/- 6.5 in 12 to 18 gestational weeks, 16.4 +/- 6.5 in 19 to 23 gestational weeks, 10.6 +/- 3.2 in 24 to 28 gestational weeks, and 5.5 +/- 1.9 in 35 to 41 gestational weeks: the mean number of nRBCs collected from 20 ml of maternal peripheral blood was 13.7 +/- 6.2. The highest value of yield was 45.6% from 12 to 18 weeks gestation. The fetal sex determination confirmed by amniocentesis or chorionic villus sampling showed 100% sensitivity and 91.7% specificity for males; 91.7% sensitivity and 100% specificity for females. We showed that fetal cells can be reliably enriched from maternal blood and that they can be used for detecting specific chromosomes by FISH with a specificity superior to current non-invasive methods. PMID- 10817029 TI - Membrane stretch increases the activity of Ca(2+)-activated K+ channels in rabbit coronary vascular smooth muscles. AB - It has been proposed that Ca(2+)-activated K+ channels play an essential role in maintaining vascular tone during stretch of blood vessel. However, the underlying mechanism of stretch-induced change of Ca(2+)-activated K+ channel activities are still unknown. The present experiment was designed to investigate the effect of membrane stretch on these channels whose activity was measured from rabbit coronary smooth muscle cells using a patch clamp technique. Ca(2+)-activated K+ channel were identified by their Ca2+ and voltage dependencies and its large conductances as in other preparations. Perfusion of cells with a hypotonic solution, which mimics stretching the cell membrane by making a cell swelling, produced an increase in channel activity in cell-attached patch mode. The similar increase was observed when negative pressure was applied into the patch pipette for stretching the cell membrane within a patch area. In inside-out patch, stretch still increased channel activity even under the conditions which exclude the possible involvement of secondary messengers, or of transmembrane Ca2+ influx via stretch-activated cation channels. Pretreatment of arachidonic acid or albumin showed no effect on stretch-induced channel activation, excluding the possibility of fatty acids mediated channel activation during membrane stretch. These results indicate that the stretch may directly increase the activity of Ca(2+)-activated K+ channels in our experimental condition. PMID- 10817030 TI - Inflammatory pseudotumor of urinary bladder. AB - A previously healthy 44-year-old male was admitted with the chief complaint of intermittent gross hematuria. On initial ultrasonographic and CT examination, a grossly protruding intravesical tumor was noted and, under the impression of a malignant bladder tumor, transurethral resection was performed. The histological findings were spindle cells with elongated cytoplasm with rare mitotic figures distributed in myxoid stroma, consistent with diagnosis of inflammatory pseudotumor of the bladder. The benign nature of this tumor warrants conservative surgical management, usually consisting of transurethral resection or partial cystectomy. No reports of metastasis have been reported following complete excision. Therefore, any suspicion and recognition of this entity is imperative to avoid performing an irreversible radical procedure. PMID- 10817031 TI - Liddle's syndrome: a report in a middle-aged woman. AB - A 54-year-old woman with diabetes mellitus was hospitalized with generalized edema and weakness. She was also found to have hypertension, hypokalemia and metabolic alkalosis. Detailed examination showed subnormal plasma renin activity and plasma aldosterone concentration. Adrenal CT scanning revealed no adrenal tumor. A successful treatment with amiloride established the diagnosis of Liddle's syndrome for the patient. Liddle's syndrome, a rare hereditary disease usually found in young patients, should be considered in the differential diagnosis of hypertension even in elderly individuals. PMID- 10817032 TI - Muscle fiber type disproportion with an autosomal dominant inheritance. AB - Congenital muscle fiber type disproportion (CFTD) has been described as a form of congenital myopathy characterized by the smallness and marked predominance of type 1 fibers in a muscle biopsy. Clinical manifestations include hypotonia, nonprogressive muscle weakness, joint contractures, and skeletal deformities. However, it has also been noted that the same pathologic alterations appeared in clinically diverse conditions. Recently, we experienced a family, a mother and two children, in which a muscle biopsy showed the mother to have muscle fiber type disproportion. This case was unusual in that there was a significant progression of weakness, an absence of neonatal hypotonia, and other commonly associated musculo-skeletal deformities. In this report, we describe the clinicopathologic features of the family with a brief review about muscle fiber type disproportion. PMID- 10817033 TI - The first Korean case of human pulmonary dirofilariasis. AB - Human pulmonary dirofilariasis has been documented from many parts of the world, but not in Korea so far. We experienced a patient of pulmonary dirofilariasis who had visited a local clinic because of chest pain for 1 month. On chest radiograph, a coin lesion of 2 cm diameter and enlargement of the mediastinal lymph node were shown. An exploratory lung resection was done. Pathologically the lesion was a pulmonary dirofilariasis complicated with necrotic pneumonia, fibrosis, and infarction. At the center of the lesion, degenerated nematode sections with multilayered cuticle, thick musculature, and bilateral internal ridges on each side were found, which was identified to be Dirofilaria immitis. This is the first report of human pulmonary dirofilariasis in Korea. PMID- 10817034 TI - Rupture of the rectosigmoid colon with evisceration of the small bowel through the anus. AB - Spontaneous rupture of the rectosigmoid colon and herniation of the small intestine through the rupture site and eventual evisceration through the anus is a very rare event. In the literature, only 42 cases have been reported. The majority of them occurred in patients with rectal prolapse and one case was reported in association with a third-degree uterine prolapse. We experienced an 81-year-old female patient with rectal prolapse and second-degree uterine prolapse complicated by spontaneous perforation of the rectosigmoid colon and anal evisceration of the small intestine. Segmental resection of the nonviable small intestine, primary repair of the ruptured rectosigmoid colon, and sigmoid loop colostomy were performed, and the patient recovered well. In our patient, both rectal and uterine prolapses cooperatively damaged the anterior wall of the rectosigmoid colon and resulted in perforation. So, rectal and uterine prolapses should be treated before the complication develops. In this patient, uterine prolapse should be treated because of the recurrence of this rare episode. PMID- 10817035 TI - Lobular carcinoma in situ in sclerosing adenosis. AB - The initial presentation of breast malignancy as noninvasive carcinoma in an area of sclerosing adenosis is unusual. Especially, lobular carcinoma in situ in sclerosing adenosis sometimes can be a potential source of confusion with invasive lobular carcinoma. We report a case of lobular carcinoma in situ presenting in adenosis exhibiting patterns akin to invasive lobular carcinoma, thus leading to potential misdiagnosis. Overall architecture of the lesion as seen at lower power and immunohistochemistry can be useful to distinguish between sclerosing adenosis with lobular carcinoma in situ and infiltrating lobular carcinoma. PMID- 10817036 TI - MRI--"magic radiotherapy imaging" for treatment planning? PMID- 10817037 TI - Imaging liver metastases: current limitations and future prospects. AB - The diagnosis of liver metastasis relies on imaging. The sensitivity of ultrasound, CT and magnetic resonance techniques for detecting liver metastases can only be assessed in comparison with surgical inspection, intraoperative ultrasound and pathological examination, all methods that are of uncertain accuracy in detecting very small lesions. With current imaging technology, we should detect virtually all liver metastases 2 cm or larger in size, and most of those 1-2 cm in size. Even with optimum imaging, at present we detect only about one-half of metastatic nodules smaller than 1 cm in patients undergoing liver resection and pathological correlation. Improvements in the earlier detection of metastases will probably require a fundamentally different approach from that of conventional anatomical methods. Micrometastases produce alterations in blood flow that may be recognized by radionuclide or Doppler perfusion methods. PMID- 10817038 TI - MRI screening for acoustic neuroma: a comparison of fast spin echo and contrast enhanced imaging in 1233 patients. AB - Gadolinium enhanced MRI is the gold standard investigation for the detection of acoustic neuroma. Non-contrast MRI sequences have been suggested as an alternative for screening examinations. In order to determine the utility of fast spin echo imaging, both gadolinium enhanced T1 weighted images and fast spin echo T2 weighted images were acquired in 1233 consecutive patients referred for exclusion of acoustic neuroma. Two radiologists independently recorded their findings. Fast spin echo T2 weighted images were evaluated with respect to the visibility of nerves within the internal auditory canals and allocated a confidence score for the presence or absence of acoustic neuroma. 33 acoustic neuromas were identified. Only 56% were confidently identified on fast spin echo T2 weighted images alone; gadolinium enhanced T1 weighted images were required to confirm the diagnosis in 44% of the cases, including 9 of the 10 intracanalicular tumours. However, when identification of two normal intracanalicular nerves is employed as the criterion of normality, the single fast spin echo T2 weighted sequence excluded acoustic neuroma in 59% of this screened population. It is concluded that an imaging strategy intended to identify small intracanalicular acoustic neuromas cannot rely on fast spin echo T2 weighted imaging alone. Gadolinium enhanced T1 weighted imaging could be restricted to patients where fast spin echo images do not exclude acoustic neuroma but this strategy requires continuous supervision by an experienced radiologist. In most practices the screening examination should continue to include a gadolinium enhanced sequence in order to optimize the detection of small acoustic neuromas. PMID- 10817039 TI - Haemostatic effects of low osmolar non-ionic and ionic contrast media: a double blind comparative study. AB - In this prospective, double-blind, randomized study the effects of a non-ionic contrast medium (Iopromide) on the haemostatic system were compared with those of a low osmolar ionic medium (Ioxaglate). The aim was to investigate in vivo whether a non-ionic contrast agent is less anticoagulant or more pro-thrombotic than an ionic medium. A large number of haemostatic parameters, including activation markers, were measured. Either Iopromide (n = 16; median volume 102 ml; 95% confidence interval 90-108 ml) or Ioxaglate (n = 15; median 105 ml; 95% confidence interval 95-114 ml) was given to 31 patients scheduled for abdominal and femoral arteriography. Blood for laboratory investigations was collected before, and 5 and 30 min after, administering the contrast medium. Indications for activation of coagulation and platelets were already found in nearly 50% of the patients before any contrast medium was given. Both Iopromide and Ioxaglate caused further increases in thrombin-antithrombin complex, prothrombin fragments 1 + 2 and beta-thromboglobulin. The degree of activation was similar for both agents. Anticoagulant effects were not observed. The haemorheological effects were compatible with haemodilution by 5-8%, again without differences between the contrast agents. Contrary to the findings from in vitro studies, we found no significant differences between the effects of the non-ionic Iopromide and the ionic Ioxaglate on coagulation and platelets. Both agents activated these systems to a limited, but identical, degree. Our results support the notion that the catheterization procedure per se may represent a source of haemostatic activation and that the ionic contrast agent studied has insufficient anticoagulant effect to prevent clotting activation being induced by the contrast medium. PMID- 10817040 TI - Distribution of cranial MRI abnormalities in patients with symptomatic and subclinical CADASIL. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare, inherited cause of early stroke and dementia, with a poor prognosis. This study was performed to clarify lesion appearance and pattern of lesion distribution in CADASIL. 20 members of a single family were tested for the CADASIL gene mutation and studied with cranial MRI. Scans were evaluated for lesion load and pattern of lesion distribution. 19 patients had cranial MRI, of whom 11 had normal MRI scans, were clinically unaffected and tested negative for the CADASIL gene mutation. The remaining eight patients had abnormal cranial MRI scans: seven patients were positive for the CADASIL gene mutation and one (untested) patient was severely clinically affected. Three of the patients who tested positive for the CADASIL gene mutation were clinically unaffected at the time of imaging. All eight patients with abnormal cranial MRI had subcortical white matter abnormalities, mostly in frontal and temporal lobes. Lesions involving the corpus callosum were present on sagittal T2 weighted images in four of five clinically affected and one of three clinically unaffected patients. Lesions involving the deep grey nuclei and the brain stem were common. On T1 weighted images, lesions were either poorly defined (confluent white matter hypointensity) or well defined (cystic infarcts or enlarged perivascular spaces). Atrophy was infrequent. Familiarity with the range of cranial MRI appearances may aid diagnosis of CADASIL. Recognition of cranial imaging features in asymptomatic CADASIL patients could prompt earlier diagnosis. PMID- 10817041 TI - Asymmetrical hemithorax volume loss due to Wegener's granulomatosis. AB - Three patients with Wegener's granulomatosis (WG) established by clinical, serological and histological criteria were noted to have marked asymmetrical hemithorax volume loss on thoracic CT. Lung dimensions were analysed from the CT in each case. Evidence of airways disease, parenchymal abnormalities and pleural changes was evaluated on CT, in order to establish the aetiology of the volume loss. Previous pulmonary infection and thoracic intervention were excluded by the clinical data. The three patients had chronic treated thoracic WG for 1-9 years. There was severe asymmetrical pleural disease in one case and parenchymal disease with evidence of fibrotic healing but no evidence of bronchial disease in two cases. Marked asymmetrical volume loss of a hemithorax is a previously unreported finding and should be added to the features of primary chronic thoracic WG. This finding does not require investigation for additional pathology. PMID- 10817042 TI - Incidence and significance of demonstrating the meniscofemoral ligament on MRI. AB - The aim was to evaluate how often MRI can detect the meniscofemoral ligament and if there is any relationship with the presence of lateral meniscal tears. Sagittal and coronal MR images of 138 patients with arthroscopically proven meniscal tears were included in this study. MR images were retrospectively reviewed for the presence of a meniscofemoral ligament and a lateral meniscal tear. The chi 2 test was used for statistical analysis. Either one or both meniscofemoral ligaments were shown in 114 (83%) of 138 patients on MR images. In 61 cases with a lateral meniscal tear, the meniscofemoral ligament was present in 82% (50/61) and absent in 18% (11/61). In 77 cases without lateral meniscal tear, the meniscofemoral ligament was present in 83% (64/77) and absent in 17% (13/77). A significant correlation was not established between the meniscofemoral ligament and lateral meniscal tear (p = 0.85). PMID- 10817043 TI - Peripheral quantitative CT of the forearm: scanner cross-calibration using patient data. AB - Bone density measurements by peripheral quantitative CT (pQCT) of the distal radius in 87 women (age 38.4 +/- 12.3 years) were obtained on a both a Stratec XCT 900 and an updated version XCT 1400. Axial area, and total and trabecular bone mineral density (BMD) results from both scanners were compared for all patients using linear regression analysis. To achieve scanner calibration we used the intercept and slope of the patients' correlations. The correlations of the patients' values were good for area (r = 0.83) as well as total BMD (r = 0.90) and excellent for trabecular BMD (r = 0.97). The cross-calibration approach in our study provided compatibility of pQCT for trabecular and total ROIs, the clinically mostly utilized volumes of measurement. PMID- 10817044 TI - Radiation doses received in the UK Breast Screening Programme in 1997 and 1998. AB - The mean glandular doses (MGDs) to samples of women attending for mammographic screening are measured routinely at screening centres in the UK Breast Screening Programme (NHSBSP). This paper reviews the data collected during screening in the NHSBSP in 1997 and 1998 for 23,752 films, using 171 X-ray sets, for 8745 women. Average MGD was 2.03 mGy per oblique film and 1.65 mGy per craniocaudal film, similar to the MGDs found previously in the NHSBSP for the years 1994 and 1995. MGD was found to increase with compressed breast thickness where the tube potential was selected manually, so that the average dose for 10 cm thick breasts was 2.7 times the average for all breasts. For large breasts (> 70 mm) the use of X-ray sets such as the IGE DMR, which automatically select the beam quality for each breast, resulted in lower doses compared with sets using manual tube potential selection. MGD to the standard breast was found to vary from 0.7 to 2.2 mGy and to be correlated with the average MGD per mediolateral oblique film for the women screened on that system (R = 0.79). No correlation between age and MGD was found within the invited age range of 50-64 years. PMID- 10817045 TI - Mandibular bone mineral density measured using dual-energy X-ray absorptiometry: relationship to hip bone mineral density and quantitative ultrasound at calcaneus and hand phalanges. AB - The main aim of this cross-sectional study was the estimation of relationships between mandibular bone mineral density (m-BMD), hip bone mineral densities (BMDs) and quantitative ultrasound at calcaneus and hand phalanges. Correlations between m-BMD and age, years since menopause (YSM) and body size were also evaluated. 42 edentulous persons (36 females and 6 males aged 60.5 +/- 6.9 years) were evaluated. In the group studied no factors affecting bone metabolism (either medical conditions or medications) were noted. Bone status was assessed by dual energy X-ray absorptiometry (mandible, hip--Lunar DPX-L), and quantitative ultrasound (calcaneus--Lunar Achilles which measures speed of sound (SOS, m s-1) and broadband ultrasound attenuation (BUA, dB MHz-1); and phalanges of the hand- DBM Sonic 1200 which measures amplitude-dependent speed of sound (AD-SOS, m s 1)). CV% for mandibular measurements was 2.06%. m-BMD correlated significantly with the following measurements: femoral neck r = 0.39, p < 0.01; Ward's r = 0.39, p < 0.01; calcaneal BUA r = 0.39, p < 0.01; and phalangeal AD-SOS r = 0.4, p < 0.01. Negative correlation consistent with a significant decrease with age was observed in m-BMD (r = -0.36, p < 0.05) and AD-SOS (r = -0.4, p < 0.01). BMD in the mandible also decreased with YSM (r = -0.47, p < 0.01). m-BMD was correlated with age, YSM, height and weight in stepwise, multiple, linear regression analysis. The following equation was obtained: m-BMD = -2.21 + 0.018 x height (cm) -0.02 x YSM (years) + 0.13 x age (years). It may be concluded that mandibular BMD may be an appropriate measurement site for the evaluation of skeletal status in osteoporosis. PMID- 10817046 TI - Radiation risk estimation in varicocele embolization. AB - Varicocele embolization is performed in healthy young men with normal life expectancy. Therefore, it is essential that the radiation risks associated with the procedure are minimized. The radiation risks associated with varicocele embolization have been estimated retrospectively from dose-area product records in a series of 41 cases and compared with a prospective series of 10 cases. Lithium fluoride dosemeters were used to measure gonad dose in the prospective series. Estimated lifetime fatal cancer risk was of the order of 0.1% in the retrospective series. A seven-fold reduction in median radiation dose was observed in the prospective series. The results indicate that with meticulous attention to technique, substantial reductions in radiation exposure can be achieved. PMID- 10817047 TI - Acute mild hypothermia caused by a low dose of X-irradiation induces a protective effect against mid-lethal doses of X-rays, and a low level concentration of ozone may act as a radiomimetic. AB - Acute changes in core body temperature following exposure to a low dose of X-rays were assessed in unanaesthetized and unrestrained mice. Radiotelemetry techniques were used to monitor core body temperature continuously. Following exposure to a 20 cGy dose of X-rays, the mice displayed a rapid and significant reduction in core body temperature relative to the sham-treated (non-irradiated) control animals. The present studies, and those by others, showed that pre-exposure to X rays at doses as low as 20 cGy may result in a reduced mortality rate following subsequent exposure to X-rays at mid-lethal dose levels. This indicates an increased tolerance to radiation. An additional experiment was conducted to examine whether the reduction in the mortality rate following exposure to mid lethal doses of radiation could be found when mice were subjected to a stressor, ozone inhalation, which induced a suppression in body temperature. The results showed that following inhalation of ozone at a concentration of 0.5 ppm, 93% of the treated animals survived a mid-lethal dose of radiation, whereas 50% of the sham-control animals died within 30 days. These results suggest that low-dose induced tolerance to radiation may be dependent on a brief exposure to ozone, and a reduction in core temperature may be necessary to obtain tolerance effects in response to a mid-lethal dose of radiation. PMID- 10817048 TI - External beam radiotherapy in exudative age-related macular degeneration: a pooled analysis of phase I data. AB - In recent years external beam radiotherapy (EBRT) has been proposed as a treatment for the wet form of age-related macular degeneration (AMD) where choroidal neovascularization (CNV) is the hallmark. While the majority of pilot (Phase I) studies have reported encouraging results, a few have found no benefit, i.e. EBRT was not found to result in either improvement or stabilization of visual acuity of the treated eye. The natural history of visual loss in untreated CNV of AMD is highly variable. Loss of vision is influenced mainly by the presenting acuity, and size and composition of the lesion, and to a lesser extent by a variety of other factors. Thus the variable outcome reported by the small Phase I studies of EBRT published to date may simply reflect the variation in baseline factors. We therefore obtained information on 409 patients treated with EBRT from eight independent centres, which included details of visual acuity at baseline and at subsequent follow-up visits. Analysis of the data showed that 22.5% and 14.9% of EBRT-treated eyes developed moderate and severe loss of vision, respectively, during an average follow-up of 13 months. Initial visual acuity, which explained 20.5% of the variation in visual loss, was the most important baseline factor studied. Statistically significant differences in loss of vision were observed between centres, after considering the effects of case mix factors. Comparisons with historical data suggested that while moderate visual loss was similar to that of the natural history of the disease, the likelihood of suffering severe visual loss was halved. However, the benefit in terms of maintained/improved vision in the treated eye was modest. PMID- 10817049 TI - A therapy beam training aid. AB - Beams of X-rays and electrons used in external beam radiotherapy are characterized by differences in penetration, build-up, divergence, scatter etc. This short communication describes a very simple phosphor-loaded box that can be used to demonstrate directly to students the different properties of beams and the effect of field modifiers such as wedge filters, lead shields etc. PMID- 10817050 TI - Investigation of dose homogeneity in paediatric anthropomorphic phantoms for a simple total body irradiation technique. AB - The technique for treating total body irradiation patients used at the centre involves no compensation for the inhomogeneity of patient shape. Dose is prescribed to the lung, and monitor units are derived from standard data depending on the external dimensions of the patient at nipple level. Dose measurements were made during standard treatments on three paediatric anthropomorphic phantoms representing children of 5, 10 and 15 years of age. The results confirmed that the measured dose to the lung was within 4% of the prescribed dose, and dose homogeneity was within +/- 5%, excluding the neck, where the higher measured doses were still within tissue tolerance. PMID- 10817051 TI - MRI of congenital coronary artery aneurysm. AB - A case is described in which a myocardial infarction in a young woman was associated with a congenital aneurysm of the left coronary artery. Coronary angiography had shown displacement and almost total occlusion of the left circumflex artery. MRI demonstrated that this was owing to an aneurysm of the left circumflex coronary artery. PMID- 10817052 TI - Tension pneumocephalus and pneumorachis secondary to subarachnoid pleural fistula. AB - A case of tension pneumocephalus and pneumorachis secondary to a subarachnoid pleural fistula after thoracic spinal surgery is described. This rare complication was diagnosed on CT. The imaging findings, significance and management of this unusual condition are discussed. PMID- 10817053 TI - Ultrasound diagnosis of oesophagostomiasis. AB - Human infection with Oesophagostomum bifurcum, a parasitic intestinal helminth, is endemic in parts of West Africa. Oesophagostomum bifurcum juveniles develop in the colonic wall, causing pus-filled granulomas. The pathology has two distinct forms. Multinodular oesophagostomiasis comprises hundreds of small nodules within a thickened, oedematous wall of the large intestine. Uninodular oesophagostomiasis, called the Dapaong tumour, presents as a painful 30-60 mm granulomatous mass in the abdominal wall or within the abdominal cavity. Diagnosis of oesophagostomiasis on clinical grounds alone is difficult. We describe cases illustrating the ultrasound appearance of these two presentations. Multinodular disease shows nodular "target" and "pseudokidney" colonic lesions. The Dapaong tumour is an echo-free ovoid lumen enveloped within a well defined poorly reflective wall. PMID- 10817054 TI - MR findings of ovarian tumours with cystic components. AB - This article depicts the MR findings of ovarian tumours with cystic components. MRI can predict the cyst contents and evaluate the potential for malignancy when an irregular solid portion or a papillary nodule is observed within the cystic tumour. Post-contrast T1 weighted MRI is helpful for the identification of solid areas that enhance. Fat suppression imaging is useful for differentiation between mature cystic teratoma and other pathology. Fat suppressed post-contrast T1 weighted MRI may demonstrate contrast enhancement, reflecting inflammatory or neoplastic processes in the pelvis. PMID- 10817055 TI - Injection of contrast media in intravenous cannulation using a power infusion device. PMID- 10817056 TI - Radiotherapy or surgery in perianal Paget's disease? PMID- 10817057 TI - A lump in the groin. PMID- 10817058 TI - Does brain penetration of anti-HIV drugs matter? PMID- 10817059 TI - Bacterial vaginosis and HIV infection. PMID- 10817060 TI - The British Co-operative Clinical Group (BCCG) PMID- 10817061 TI - The repertoire of human efforts to avoid sexually transmissible diseases: past and present. Part 1: Strategies used before or instead of sex. AB - BACKGROUND/OBJECTIVE: Despite the focus by public health programmes on condoms, chastity, or monogamy, people use a much wider variety of strategies to minimise their personal risk of sexually transmissible disease (STD). The objective of this study was to compile a comprehensive list of personal and societal STD avoidance strategies. METHODS: Data from clinical and research observations, computer searches, and historical texts were pooled. RESULTS: In addition to discriminating between potential sexual partners, a variety of behaviours before or instead of sex were identified that have been perceived to alter STD risk. Traditional STD avoidance strategies were often poorly documented and difficult to disentangle from other drives such as the maintenance of social order, paternity guarantee, and eugenics. They also varied in popularity in time and place. Some examples were displacement activities such as masturbation or exercise, circumcision, infibulation, shaving, vaccination, or requiring partners to be tested for infection. Social and moral forces typically discourage non marital sex, and this affects most people most of the time but few people all of the time. CONCLUSION: The full spectrum of STD avoidance strategies warrants further study because some are ubiquitous across cultures and because they have the potential to complement or undermine safer sex programmes. Because of their greater acceptability, some less efficacious strategies may have greater public health importance than less popular but more efficacious strategies such as condoms. PMID- 10817062 TI - Attack rates of human papillomavirus type 16 and cervical neoplasia in primiparous women and field trial designs for HPV16 vaccination. AB - BACKGROUND: Identification of human papillomavirus type 16 (HPV16) as the major risk factor for cervical neoplasia, and mass production of DNA free HPV capsids have paved the way to preventive vaccination trials. Design of such trials requires reliable attack rate data. OBJECTIVE: Determination of (1) HPV16 and (2) cervical neoplasia attack rates in primiparous women. Estimation of actuarial sample sizes for HPV16 vaccination phase IV trials. DESIGN: A longitudinal cohort study. METHODS: Population based Finnish Maternity Cohort (FMC) and Finnish Cancer Registry (FCR) were linked for the identification of two cohorts of primiparous women: (1) a random subsample of the FMC: 1656 women with two pregnancies between 1983-9 or 1990-6 and living in the Helsinki metropolitan area, and (2) all 72,791 primiparous women living in the same area during 1983 94. Attack rate for persistent HPV16 infection (1) was estimated in 1279 seronegative women by proportion of seroconversions between the first and the second pregnancy. Comparable 10 year cumulative incidence rate (CR) of cervical intraepithelial neoplasia grade III and cervical cancer (CIN III+) (2) was estimated based on cases registered at the FCR during 1991-4. RESULTS: The HPV16 attack rates were 13.8% (< 18 years), 7.0% (18-19 years), 2.3% (21 years), 2.4% (23 years), and 4.5% (< 25 years). Number of vaccinees required for a 5 year efficacy trial with persistent HPV16 infection as the end point ranged between 1000 and 3900, assuming 80% power, 90%-70% vaccine efficacy (VE), and misclassification. The CRs of CIN III+ were 0.33% (< 18 years), 0.44% (18-19 years), 0.21% (20-24 years), and 0.28% (< 25 years). Number of vaccinees required for a 10 year efficacy trial with HPV16 positive CIN III+ as the end point was 15,000 assuming 80% power, 90% VE, and 75% aetiological fraction of CIN III+ for HPV16. CONCLUSIONS: The attack rates of HPV16 and CIN III+ identify primiparous women under 25 years of age among target populations for postnatal HPV vaccination at phase II/III trials. PMID- 10817064 TI - Home screening for chlamydial genital infection: is it acceptable to young men and women? AB - OBJECTIVES: To determine the acceptability, to young men and women, of home screening for chlamydial infection. METHODS: We wrote to a random sample of 208 women aged 18-25 years and 225 men aged 18-35 years from three general practices, inviting them to undergo home screening for chlamydial infection. They were asked to return, by normal post, a urine specimen (for men and half of the women) or a vulval swab (other half of the women) for ligase chain reaction (LCR) testing for chlamydial infection. They were also asked to return a short questionnaire about risk status and the acceptability of this approach. RESULTS: The participation rate among the available sample was 39% for women and 46% for men (p = 0.3). However, among women, the rate was slightly higher (p = 0.05) for urine samples (47%) than for vulval swabs (32%). Six per cent of women and 9% of men declined to take part, while 42% of women and 33% of men failed to respond. Two men objected to receiving the package at home. We received few other comments, positive and negative in about equal measure. CONCLUSION: Home screening for chlamydial infection is a potentially efficient method of reaching young people who may have little contact with health services. Men were at least as likely as women to respond to this screening approach. Home screening might form a useful component of a future chlamydial screening programme in the United Kingdom. PMID- 10817063 TI - Investigation of the increased incidence of gonorrhoea diagnosed in genitourinary medicine clinics in England, 1994-6. AB - OBJECTIVES: To determine important risk factors associated with cases of gonorrhoea in England, and whether any particular risk groups were associated with the substantial rise in numbers of cases seen between 1994 and 1996. DESIGN: Two retrospective cross sectional surveys. SETTING: 70 randomly selected genitourinary medicine (GUM) clinics in England. SUBJECTS: 10% of all gonorrhoea patients attending GUM clinics in England in 1994 (847 patients) and 1996 (1146 patients). MAIN OUTCOME MEASURES: For risk factors in 1996 (study 1), unadjusted rates per 100,000 population aged 14-70 and relative rates (RR) with 95% confidence intervals (CIs). For the change in risk factors between 1994 and 1996 (study 2), adjusted odds ratios (ORs) with 95% CIs, derived from logistic regression analyses of data on patients in 1996, with patients in 1994 as the comparison group. RESULTS: The incidence of gonorrhoea in 1996 was higher in homosexual males (812 per 100,000; RR = 30.2, CI = 25.2 to 36.0) compared with heterosexual males (27 per 100,000); in black Caribbeans (467 per 100,000; 21.4, 17.9 to 25.5) and black Africans (235 per 100,000; 10.8, 7.5 to 15.5) compared with white people (22 per 100,000); and in previous GUM clinic attenders (433 per 100,000; 37.93, 35.46 to 40.56) compared with those who had not attended previously (11 per 100,000). However, most patients were either white or heterosexual. Heterosexual patients in 1996 were significantly more likely to have reduced sensitivity to penicillin (2.55, 1.20 to 5.41) than those in 1994. Male homo/bisexual patients in 1996 were significantly more likely to be from the north west (3.77, 1.45 to 9.80) and to have either reduced sensitivity (2.63, 1.03 to 6.73) or complete resistance (1.98, 1.03 to 3.78) to penicillin, compared with those in 1994. CONCLUSIONS: Homo/bisexual men and the black Caribbean population in England experience a disproportionate burden of gonococcal infections, however, the bulk of diagnoses are in white heterosexuals. No single risk group was associated with the rise in numbers of cases between 1994 and 1996. Resistance to penicillin is widespread and has increased in homo/bisexual men, and it is possible that a rise in treatment failures has, to some extent, enhanced transmission of gonorrhoea and contributed to the rise in numbers of diagnoses in this group. PMID- 10817065 TI - Epidemiology of ectopic pregnancy during a 28 year period and the role of pelvic inflammatory disease. AB - OBJECTIVES: We analysed the epidemiology of ectopic pregnancy (EP) during a 28 year period, 1970-97, using methods applicable to ecological studies in order to test the hypothesis that a reduction of pelvic inflammatory disease (PID) will be associated with a decrease of EP. METHODS: Hospital records of patients aged 15 54 admitted to our department of gynaecology were reviewed for EP and PID for the period 1 January 1970 to 31 December 1997. EP for the period 1970-4 was based on available statistics. The total number for EP was 1270 and for PID 2559. The total population for the catchment area was 100,000-120,000 during the study period. Incidences were age standardised and calculated using official population statistics to represent the average female population in the five 5 year periods 1970-4, 1975-9, 1980-4, 1985-9, 1990-4, and in each of the consecutive years 1995, 1996, and 1997. Incidences for EP were calculated per 1000 women and per 1000 pregnancies while those for PID per 1000 women. National statistical data of EP were available for 1975-94 and were used for comparison with the local study. RESULTS: The EP incidences increased from 7.7 per 1000 pregnancies in the first 5 year period to 13.4 in the second, and continued to rise for another decade reaching the peak figures of 16.6 in 1985-9--that is, more than a twofold increase. Since then and to 1997 the EP incidence has decreased by 30%. PID admissions increased during the study period from 2.7 per 1000 women in the first 5 year period to 3.2 in the second. From then on they continuously decreased and reached a low of 0.5 in 1997. The greatest changes occurred in women < or = 24 years of age. The peak incidence for this age group was 7.7 in 1975-9, and the lowest was 0.4 per 1000 women in 1996. The greatest reduction of EPs was noted for women < or = 24 years old, from a high of 10.0 in 1975-9, coinciding with the peak incidence of PID, to a low of 4.0 in 1997, a reduction of 58.4%. The incidence of EP was two to three times higher in women > or = 25 years old, most obvious in those > or = 30 years, with peak figures of 20.9 per 1000 pregnancies in 1985-9, and 13.9 in 1997, a reduction of 33.4% and the lowest figures for the past 23 years. For women aged 25-29 years the incidence peaked in the previous 5 year period 1980-4--that is, one 5 year period later than for those < or = 24 years, which we interpret as cohort effects in relation to PID. CONCLUSIONS: Reduction of PID was strongly associated with a decline of EP. The decline was greater and immediate for women < or = 24 years old, than for those > or = 25 years. The two to three times higher EP incidence in women > or = 25 years of age was most probably due to a cohort effect as the peak of PID occurred a decade earlier in women < or = 24 years old. Prevention of PID may not only reduce EP but also reduce adverse effects on tubal patency. PMID- 10817066 TI - Validity of the vaginal discharge algorithm among pregnant and non-pregnant women in Nairobi, Kenya. AB - OBJECTIVE: To evaluate the validity of different algorithms for the diagnosis of gonococcal and chlamydial infections among pregnant and non-pregnant women consulting health services for vaginal discharge in Nairobi, Kenya. METHODS: Cross sectional study among 621 women with complaints of vaginal discharge in three city council clinics between April and August 1997. Women were interviewed and examined for symptoms and signs of sexually transmitted infections (STIs). Specimens were obtained for laboratory diagnosis of genital infections, HIV, and syphilis. The data were used to evaluate the Kenyan flow chart as well as several other generated algorithms. RESULTS: The mean age was 24 years and 334 (54%) were pregnant. The overall prevalence rates were: 50% candidiasis, 23% trichomoniasis, 9% bacterial vaginosis, 7% gonorrhoea, 9% chlamydia, 7% syphilis, and 22% HIV. In non-pregnant women, gonococcal and chlamydial infection was significantly associated with (1) demographic and behavioural risk markers such as being single, younger than 20 years, multiple sex partners in the previous 3 months; (2) symptom fever; and (3) signs including presence of yellow or bloody vaginal discharge, cervical mucopus, cervical erythema, and friability. Among pregnant women only young age, dysuria, and fever were significantly associated with cervical infection. However, none of these variables was either sensitive or specific enough for the diagnosis of cervical infection. Several algorithms were generated and applied to the study data. The algorithm including risk markers performed slightly better than the current Kenyan algorithm. CONCLUSION: STIs form a major problem in the Nairobi area and should be addressed accordingly. None of the tested algorithms for the treatment of vaginal discharge would constitute a marked improvement of the existing flow chart. Hence, better detection tools for the specific aetiology of vaginal discharge are urgently needed. PMID- 10817067 TI - Sexually transmitted infections and HIV in a rural community in the Lesotho highlands. AB - OBJECTIVES: To conduct a knowledge, attitude, and practice (KAP) study and to determine the prevalence of sexually transmitted diseases (STDs), including HIV, in a community residing in remote, rural Lesotho. METHODS: In 1995 a cross sectional, community based epidemiological study was conducted on a population of 7500 people living in 89 villages. A total of 29 villages were randomly selected and a systematic sample of houses within villages was obtained. Questionnaires were administered to subjects after written consent was obtained. Determination of N gonococcus and C trachomatis infection was done on urine using ligase chain reaction (LCR) technology. Using blood specimens, syphilis was diagnosed by RPR and TPHA tests and HIV by a single ELISA and confirmed with a western blot. RESULTS: Questionnaires were administered to 277 women, 100 men, and 149 youths (12-15 years). Chlamydia was diagnosed in 28.4% of adults, gonorrhoea in 5.9%, syphilis in 11.3%, and HIV infection in 6.3%. All cases of HIV occurred along the main road (p = 0.001) and 72% of individuals with gonorrhoea were co-infected with chlamydia (p = 0.0001). 11.6% of women and 38.0% of men had had sex with a non-regular partner in the past 3 months and none had used condoms. CONCLUSION: A high prevalence of STDs and HIV infection was found in a population characterised by low levels of knowledge about STD/HIV, high risk sexual behaviour, and evidence of inappropriate health seeking behaviour for STDs. PMID- 10817068 TI - HIV associated culture proved tuberculosis has increased in north central London from 1990 to 1996. AB - OBJECTIVES: To examine rates of culture proved tuberculosis in HIV infected patients in three specialist centres in north central London. METHODS: Cases of tuberculosis in patients with previously documented HIV infection from 1990 to 1996 were identified retrospectively from microbiological/clinical records at Chelsea and Westminster, St Mary's, and University College London Hospitals. RESULTS: Between 1990 and 1996 202 cases of culture proved tuberculosis were identified at the three centres. Of these, 132/202 (65.3%) occurred in homosexual/bisexual men, 41/202 (20.3%) were in patients with heterosexual contact in sub-Saharan Africa, and 29/202 (14.4%) were in "others." Overall 148/202 (73.3%) had pulmonary tuberculosis. The total number of HIV infected individuals seen at the three centres increased from 4298 in 1990 to 5048 in 1996. Rates of tuberculosis in the three centres increased from 0.46% in 1990 to 0.83% in 1996. Part of this increase was due to an increase in tuberculosis among Africans from 1993 to 1996. CONCLUSIONS: Rates of HIV associated tuberculosis increased in these three centres in north central London between 1990 and 1996. In part this was due to an increase in the number of African patients with HIV infection attending the three centres. In addition, there was circumstantial evidence of recent transmission among homosexual men with HIV infection. Prospective "real time" surveillance of tuberculosis in HIV infected patients is needed in order to detect case clustering and to improve tuberculosis control. PMID- 10817069 TI - A new visual indicator of chlamydial cervicitis? AB - OBJECTIVES: To determine the usefulness of endocervical discharge opacity as a risk indicator for chlamydial infection in relation to two acknowledged visual indicators--yellow endocervical discharge and easily induced mucosal bleeding of the cervix. METHODS: Women from two family planning clinics, a therapeutic abortion clinic, and a university student health clinic (n = 1418 total) consented to a pelvic examination and chlamydia testing, and completed a questionnaire on socio-demographics, sexual behaviour, medical history, and symptoms. A case of chlamydia was defined as positive by culture or blocked enzyme immunoassay in an endocervical swab. RESULTS: The prevalence of chlamydial infection in the clinics was 6.3%. All three of the visual indicators--yellow endocervical discharge, easily induced bleeding, and opaque cervical discharge- were statistically significantly and independently associated with chlamydial infection (odds ratios 2.8, 2.3, and 2.9 respectively), independent of clinic type. Adjustment for the other visual indicators made little difference to the odds ratios. CONCLUSION: Opacity of endocervical discharge was at least as important as the other two commonly acknowledged indicators of chlamydial cervicitis--yellow endocervical discharge and easily induced mucosal bleeding of the cervix. PMID- 10817070 TI - Psychological consequences of sexual assault among female attenders at a genitourinary medicine clinic. PMID- 10817071 TI - Multiple, genital lobular capillary haemangioma (pyogenic granuloma) in a young woman: a diagnostic puzzle. AB - A 21 year old woman presented with multiple lobulated lesions on the labia majora. The surface of most of the lesions was ulcerated revealing a glistening surface. All lesions were excised. The histopathology revealed features suggestive of lobular capillary haemangioma (pyogenic granuloma). Pyogenic granuloma is considered as a reactive hyperproliferative vascular response to trauma or other stimuli. A literature search revealed reports of a few cases of lobular capillary haemangioma of the glans penis but not on the female genitalia. This case is presented to help physicians become aware that lobular capillary haemangiomas (pyogenic granuloma) may occur at this site. PMID- 10817072 TI - Syphilis screening programme in Athens, 1974-98. PMID- 10817073 TI - Extrapulmonary tuberculosis masking disseminated histoplasmosis in AIDS. PMID- 10817074 TI - "Sidies". PMID- 10817075 TI - Prevalence of HPV cervical infections among imprisoned women in Barcelona, Spain. PMID- 10817076 TI - Detection of penicillinase producing Neisseria gonorrhoeae strains in Cuba, 1995 8. PMID- 10817077 TI - Rising HIV prevalence in STD clinic attenders at Chandigarh (north India)--a relatively low prevalence area. PMID- 10817078 TI - HIV seropositivity in women with syphilis in Delhi, India. PMID- 10817079 TI - Immune reconstitution CMV pneumonitis. PMID- 10817081 TI - Plasminogen activator inhibitor-1 and carotid intima-media thickening in patients with newly detected primary hypertension. AB - OBJECTIVE: To investigate the correlation between ultrasonographically evaluated intima-media thickness (IMT) of common carotid artery (CCA) and cardiovascular risk factors for subjects with newly detected, uncomplicated and untreated primary hypertension. METHODS: The study population consisted of 200 subjects (123 men and 77 women, aged 46+/-7.5 years). Blood pressure was measured in the clinical setting and by 24 h noninvasive ambulatory monitoring. Fasting levels of blood glucose, plasma lipids and lipoproteins, fibrinogen and plasminogen activator inhibitor (PAI)-1 were measured. Ultrasound examination included measurement of far-wall intima-media complex of CCA and morphologic evaluation of occurrence of plaques in carotid and femoral bifurcations. RESULTS: The prevalence of greater than normal IMT (mean IMT > or =0.80 mm) was 22%. Significant univariate correlations to the dichotomy between normal and greater than normal mean IMT were detected for age, smoking, level of LDL cholesterol, level of PAI-1 and total ultrasonographic score. Multivariate logistic regression analysis confirmed the associations between greater than normal mean IMT and plasma concentrations of LDL cholesterol and PAI-1 as well as total ultrasonographic score. CONCLUSION: Greater than normal IMT of CCA was more strictly related to other cardiovascular risk factors than it was to blood pressure and was strongly associated with the occurrence of atherosclerotic plaques in carotid and femoral arteries. The role of PAI-1 in intima-media thickening that is emerging suggests that fibrinolytic balance is an important determinant of vessel-wall homeostasis in hypertensive patients. PMID- 10817080 TI - Clustering of biological risk factors for cardiovascular disease and the longitudinal relationship with lifestyle of an adolescent population: the Northern Ireland Young Hearts Project. AB - OBJECTIVE: To determine whether clustering of biological cardiovascular disease (CVD) risk factors exists and to investigate the longitudinal relationship between lifestyle parameters (dietary intake, physical activity and smoking behaviour) and clustering of biological CVD risk factors, which was defined as belonging to one or more sex-specific 'high-risk' quartiles for the ratio between levels of total serum cholesterol and high-density lipoprotein cholesterol, diastolic blood pressure, body fatness (sum of four skinfolds) and cardiopulmonary fitness (number of laps accomplished on a 20 m shuttle-run test). METHODS: The study was part of the Northern Ireland Young Hearts Project, a 3 year observational longitudinal study concerning adolescent boys (n=229) and girls (n=230) who were initially aged 12 years. The longitudinal relationships were analysed with generalized estimating equations. RESULTS: Significant clustering of biological CVD risk factors was observed both for boys and for girls, but the stability over time was rather low. Smoking was the only lifestyle parameter related to this clustering and was observed only among girls (rate ratio 1.5, P < 0.01); furthermore, none of the lifestyle parameters was significantly related to this clustering. CONCLUSIONS: Because biological CVD risk factors tend to cluster, it is important to investigate these risk factors together. However, for subjects in this age group, according to our analysis, lifestyle parameters were hardly related to this clustering. PMID- 10817082 TI - Plasma level of homocysteine is correlated to extracranial carotid-artery atherosclerosis in non-hypertensive Japanese. AB - BACKGROUND: Results of some epidemiologic studies in Western countries have clarified that hyperhomocysteinemia is a plausible risk factor for atherosclerotic vascular disease, but its role in Japanese communities is not known. DESIGN: A community-based cross-sectional design. METHODS: We performed a cross-sectional study of 474 elderly men aged 60-74 years in two Japanese rural communities (Noichi in southwestern Japan and Ikawa in northeastern Japan). We examined the association between plasma concentrations of homocysteine and the maximum intima-media thickness (assessed by ultrasonography). RESULTS: The prevalence of thickening was 10.7% for the lowest tertile of homocysteine level and 21.1% for the highest tertile. For the subjects without hypertension, the odds ratio for having carotid intima-media thickening was 5.8; it was significantly higher for the highest tertile of homocysteine level than it was for the lowest after adjusting for age, hypercholesterolemia, hypoalphalipoproteinemia, diabetes, and smoking by using a multiple logistic regression model. However, its correlation was not evident for those with hypertension. CONCLUSIONS: High levels of plasma homocysteine are correlated to extracranial carotid artery atherosclerosis in elderly men without hypertension in Japanese rural communities. PMID- 10817083 TI - Job control, job demands and social support at work in relation to cardiovascular risk factors in MONICA 1995, Goteborg. AB - BACKGROUND: Job control and work environment are related to risk of coronary heart disease (CHD), but there is limited understanding of the independent risks associated with these factors. OBJECTIVE: To investigate the association between psychosocial work characteristics and biological risk factors for both sexes for a random population sample in Goteborg, Sweden. DESIGN: A cross-sectional study. METHODS: We used an age-stratified random sample of men and women aged 25-64 years comprising 1200 men and 1412 women, from which 746 men and 872 women responded to the invitation for screening, which included questionnaires and physical/laboratory investigations in 1995. RESULTS: Women had lower job control than did men (P=0.00001); job demands were equal and social support at work slightly higher among women (P=0.04). Job control was positively related to education and social group. Smoking women had low job control and high job demands. Women with high grades of psychological stress had low job control and low social support at work (P=0.001 and P=0.01). For both sexes job demands were high (P=0.0001) among those who reported high psychological stress. Men with high job control and high social support at work were more physically active during leisure time. Subjects with job strain had low social support (P=0.01). Job stress factors were not related to biological coronary risk factors. CONCLUSIONS: Women had lower job control than did men. Job control was positively related to education, social class and physical activity. Psychosocial factors were not related to biological coronary risk factors. PMID- 10817084 TI - Risk factors for coronary heart disease and persistent infection with Chlamydia pneumoniae or cytomegalovirus: a population-based study. AB - BACKGROUND: A large number of epidemiological and pathological studies have reported on associations between coronary heart disease and persistent infection with Chlamydia pneumoniae or cytomegalovirus, but relatively few have reported on possible relations between these infections and vascular risk factors. OBJECTIVE: To determine whether serum concentrations of immunoglobulin G antibodies to C. pneumoniae or cytomegalovirus are correlated to standard vascular risk factors, markers of inflammation and indicators of socioeconomic status. METHODS: We performed a cross-sectional sero-epidemiological study nested within a randomized trial involving five general practices in Bedfordshire, UK. We made measurements of a number of standard vascular risk factors, serum markers of systemic inflammation and other relevant characteristics in 704 individuals. RESULTS: There were significant associations between C. pneumoniae immunoglobulin G levels and male sex and cigarette smoking (2P < 0.01 for each) and between cytomegalovirus immunoglobulin G levels and age (2P < 0.001). Other factors were not significantly associated with serum antibodies to either persistent infection. CONCLUSIONS: Serological evidence of persistent infection with C. pneumoniae or cytomegalovirus in this population was not strongly associated with most standard vascular risk factors and other characteristics. The main implication is that such risk factors are not likely to be important confounders or mediators of the reported associations between coronary heart disease and these agents. PMID- 10817085 TI - Social relations and the metabolic syndrome in middle-aged Swedish women. AB - BACKGROUND: Both social isolation and the metabolic syndrome are independently associated with greater than normal cardiovascular risk. DESIGN: A population based cross-sectional study of middle-aged Swedish women. METHODS: The study group consisted of 300 healthy women (aged 31-65 years) who were representative of women living in the greater Stockholm area. Social isolation was measured by using a condensed version of the Interpersonal Support Evaluation List. Health behaviours were assessed and a full serum-lipid-level and haemostatic profile was obtained by standardized methods. The metabolic syndrome was defined as the presence of two or more of these components: fasting serum level of glucose > or = 7.0 mmol/l, arterial blood pressure > or = 160/90 mmHg, fasting serum level of triglycerides > or = 1.7 mmol/l or high-density lipoprotein < 1.0 mmol/l, or both, and central obesity (waist:hip ratio > 0.85 or body mass index > 30 kg/m2, or both). RESULTS: After adjustment for age, menopausal status, educational level, smoking, exercise habits and consumption of alcohol, the risk ratio for the metabolic syndrome for women in the lower compared with women in the upper social-support quartile was 3.5 (95% confidence interval 1.1-11.4), whereas that of women in the two middle quartiles was 2.2 (95% confidence interval 0.67-7.2; P for trend 0.02). CONCLUSIONS: Social isolation was associated with the metabolic syndrome for these middle-aged women. The findings suggest that the metabolic syndrome and its components may be mediators of the reported association between social isolation and cardiovascular disease. PMID- 10817086 TI - Efficacy and safety of cerivastatin for type 2 diabetes and hypercholesterolaemia. Hyperlipidaemia in Diabetes Mellitus investigators. AB - BACKGROUND: The prevalence of coronary heart disease (CHD) is markedly increased in diabetic patients compared with non-diabetic individuals, and its prognosis is less good. Serum total and low-density lipoprotein (LDL) cholesterol concentrations have been shown to be powerful predictors of CHD morbidity and mortality in patients with type 2 diabetes. The available data suggest that the target cholesterol concentration in patients with diabetes should be similar to that in non-diabetic individuals with a previous myocardial infarction. This led us to investigate the efficacy, tolerability and safety of a new, highly potent statin, cerivastatin, in diabetic hyperlipidaemia. METHODS: This was a multinational, multicentre, double-blind, randomized study in type 2 diabetic patients with hypercholesterolaemia (LDL cholesterol >3.35 mmol/l; triglycerides <4.56 mmol/l). Eligible patients were randomly assigned to groups to receive cerivastatin 0.1 mg or 0.3 mg or placebo in a ratio of 2:2:1 for 12 weeks. They were monitored in the clinic every 4 weeks. RESULTS: Of the 453 patients screened, 265 were allocated to the study groups. Fifty-one received placebo and 107 patients were assigned to each active treatment group (0.1 mg and 0.3 mg cerivastatin). At the close of the study, total cholesterol had decreased by 13.7% and 23.5%, LDL cholesterol decreased by 20.2% and 33.8%, and triglyceride concentrations decreased by 3.9% and 12.3% in the cerivastatin 0.1 mg and 0.3 mg groups, respectively. There was no significant difference between the groups in haemoglobin A1c, adverse events or increases in liver and muscle enzymes during the study period. CONCLUSIONS: Hypercholesterolaemic patients with type 2 diabetes had a significant reduction in LDL cholesterol and total cholesterol concentrations after cerivastatin treatment once daily. The dose of 0.3 mg cerivastatin is effective in diabetic hypercholesterolaemia, with co-reduction of triglyceride concentrations. The effect of cerivastatin on coronary morbidity and mortality is currently being investigated in clinical trials. PMID- 10817087 TI - Bibliography. Current world literature. PMID- 10817088 TI - Improving reproductive sexual health: a primary goal at the beginning of the new millennium. AB - At the beginning of the millennium, it is important that experts and official organizations direct their strategic plans towards the improvement of reproductive sexual health. Adolescents represent the major target of these plans in an effort to improve their sexual behavior. Sexually transmitted diseases must be reduced since their prevalence is very high in certain eastern European countries. These plans should be considered together with the development of new fertility control methods which provide at the same time bactericidal and antiviral properties. Education, provided especially through the media, should be considered a tool for the improvement of the present situation. PMID- 10817089 TI - From the contraceptive to the anthropocentric revolution (Gregory Pincus in memoriam). AB - The author has seen more progress in science and technology than all scientists of all preceding periods together since the dawn of history and has witnessed a number of powerful revolutions (e.g. scientific, information, postindustrial, globalization, environmental, contraceptive, reproductive health, gender equity and demographic) that have profoundly changed the world and our perception of it. The contraceptive revolution started off this irreversible process with spectacular results. By 1990, contraception had become a global lifestyle for more than 900 million couples. It also resulted in subsequent revolutions in reproductive health and gender equity in a new world with a dramatically changed population structure with fewer and fewer children and more and more elderly. These revolutions will assist humankind in the 21st century in deconstructing the deterministic world view of past centuries and replacing it with a science-driven anthropocentric world view. PMID- 10817090 TI - Family planning perspectives at the beginning of the next century. AB - Looking back over the past 40 years or so, family planning has indeed been the success story of development; it has made an extraordinary difference to the lives of tens of millions of women. Globally speaking, the unmet needs for family planning and reproductive health still exist. Meeting these challenges would require better and improved methods of contraception, addressing issues such as sexually transmitted infections, unsafe abortions, cultural, religious, legal and political barriers. Better utilization of communication channels and investment in 'human capital'--informing people, opening a dialogue to reach consensus for action, are crucial. Finally, changes at policy and administrative level, including improvements in financial and human resource management, and making family planning and reproductive health programs sustainable will be important as we move into the 21st century. PMID- 10817091 TI - Reproductive health for all: a commitment towards the new millennium. AB - The world government community made a commitment in 1994 to make reproductive health care universally available no later than the year 2015. The commitment has been confirmed by a special session of the United Nations General Assembly in July 1999. The commitment has to be backed up by resources, and the health profession has to provide the know-how. Social action will also be needed. Progress has been made, but challenges remain. PMID- 10817092 TI - Reproductive and sexual health and safe motherhood in the developing world. AB - The aim of this paper is to discuss the main hurdles and possible solutions concerning reproductive and sexual health and safe motherhood in the developing countries. In response to the changing global situation, a broader new concept of reproductive health has emerged. The Programme of Action of The International Conference on Population and Development (ICPD) emphasizes and highlights various aspects of reproductive health. No population in the world has met the goals of the ICPD. The problems are particularly acute in developing countries: between 20 and 40% of births are unwanted or wrongly timed, posing hardships for families and jeopardizing the health of a million women and children. An estimated 50 million induced abortions are performed each year, with some 20 million of these performed under unsafe conditions or by untrained providers. Almost 600,000 women die every year due to pregnancy-related causes, 99% of them in developing countries. Approximately 7.6 million infant deaths occur during the perinatal period each year. There are more than 333 million new cases of curable sexually transmitted diseases world-wide each year. Nearly 22 million people are estimated to be infected with the human immunodeficiency virus, of whom 14 million are in sub-Saharan Africa, with rapidly increasing numbers of infected persons in South and South-East Asia. When compared to the developed world, these figures are staggering because of barriers operating at different levels and preventing women from receiving care that is timely and of good quality. United efforts of obstetricians, pediatricians, sociologists, and different governmental and non governmental organizations are required to achieve our targets. PMID- 10817093 TI - From abortion to contraception in Europe. AB - OBJECTIVES: This review summarizes abortion statistics, mainly in Europe and primarily in Eastern Europe. The reasons for unwanted pregnancies and consequent abortions are analyzed. Recommendations are proposed to achieve a shift from abortion to contraception in fertility regulation. METHODS: The relevant literature and the results of the data collected by the author in 16 Eastern European countries by means of a questionnaire are reviewed. CONCLUSIONS: The need and the possibilities for better future perspectives are outlined in the conclusions of The Szeged Declaration. The major problems identified were the low level or lack of training of professionals, deficiencies in the knowledge of current family planning methods among professionals, consumers, policy makers and media representatives, the lack of supplies of modern contraceptives to meet the needs of the population, the lack of sex education in schools and the high level of induced abortions. In the majority of the countries in the Eastern European region, there is a need for the establishment of a system for the delivery of high-quality family planning services. This imposes a requirement for the establishment of a system for the continuous supply of contraceptives, through national family planning programs, by governmental and/or nongovernmental organizations. To increase contraceptive use, countries in this region should consider the establishment of extensive educational programs on current family planning methods and other aspects of reproductive health for their health professionals, consumers and representatives of the media. Sex education in schools should be promoted. Governments should be encouraged to invest more in setting up and running family planning services and programs. PMID- 10817094 TI - Guaranteeing freedom of choice in matters of contraception and abortion in Europe: some personal remarks. AB - The most important beneficial effect of modern contraception is that it has promoted the concept of birth control as a fundamental right of women from the viewpoint of equality between men and women. This right will lead to further rights for women, in particular, that of a free choice in respect of their sex lives, their method of procreation and, in more general terms, their way of life and their legitimate place in society. PMID- 10817095 TI - Contraceptive services for adolescents in Latin America: facts, problems and perspectives. AB - This review presents facts about sexual and contraceptive behavior of Latin American adolescents, analyzes barriers to contraception, and summarizes present perspectives. Between 13 and 30% of Latin American adolescent women live in union before their 20th birthday and between 46 and 63% have had sexual relations. The prevalence of contraceptive use among adolescents at risk of pregnancy remains very low. The pill is the best known contraceptive method. When sexual activity becomes a permanent practice, contraceptive use increases but remains low. Barriers to contraception can be identified as: (1) arising from adolescents themselves (moral objections, alleged medical reasons, lack of confidence in adults and in the health system, promiscuity; (2) arising from the sexual partner (partner's opposition, masculine irresponsibility); (3) arising from adults (moral objections, fear of sex education, adult control and power of decision making); (4) arising from the health system (inappropriateness of services, regulatory barriers, gender inequality); (5) arising from health professionals (medical barriers to contraceptive use, discomfort with sexual matters); (6) arising from the educational system (educational failure, teachers' reluctance); and (7) arising from other social agents (religious opposition, media ambivalent messages, fund restraints). There have been improvements in recent years, including the achievements of groups working for and with adolescents, and the support from distinguished personalities. PMID- 10817096 TI - Contraceptive strategies for young women in the 21st century. AB - Safe, effective and affordable contraceptives have been available for a few decades, yet, in many countries, the struggle for reproductive rights continues. Children, still, are forced to give birth to children in many nations. In most industrialized countries, where contraceptive counselling and abortions have been options, fertility rates have reached all time lows. Effective contraception improves health and well-being as well and may be used for conditions other than birth control. Young girls often initially take oral contraceptives primarily to reduce menstrual pain and blood loss; they also welcome a contraceptive that eliminates menstrual bleeding. Women using oral contraceptives and Norplant experience about 50% reduction in menstrual blood; 90% reduction in blood loss is achieved with the levonorgestrel intrauterine system (Mirena). Teenagers and their parents are often misinformed about the side-effects of oral contraceptives and birth control in general, which may adversely affect compliance. Adequate, user-friendly and supportive information about contraception is necessary to ensure proper use of the pill and other birth control methods. In addition, emergency contraceptives should be easily accessible. The mortality and morbidity of young women would be dramatically reduced by the global use of medical abortion as well. Only a combined effort by policy makers, educators, parents and health-care providers can enhance the reproductive (and future!) health of both young females and males. PMID- 10817097 TI - Appropriate clinical management of women taking valproate. PMID- 10817098 TI - Blue genes and the mechanism of action of antidepressants. PMID- 10817099 TI - Positive treatment effect of estradiol in postpartum psychosis: a pilot study. AB - BACKGROUND: Postpartum illnesses with psychiatric symptoms and serious adverse sequelae are highly prevalent during the childbearing years. Despite multiple medical contacts, these illnesses often remain unidentified and untreated. To study the association between estradiol and puerperal psychosis, we measured serum concentration of estradiol and performed an open-label trial of physiologic 17beta-estradiol in women with this disorder. METHOD: Ten women with ICD-10 psychosis with postpartum onset consecutively recruited from a psychiatric duty unit were studied. Serum estradiol concentration was measured at baseline and weekly during sublingual 17beta-estradiol treatment for 6 weeks. The treatment effect was evaluated by a clinician-rated psychiatric symptom scale (the Brief Psychiatric Rating Scale [BPRS]). RESULTS: The baseline serum estradiol levels (mean = 49.5 pmol/L; range, 13-90 pmol/L) were even lower than the threshold value of gonadal failure, and the patients exhibited high scores on the psychiatric symptom scale (mean BPRS total score = 78.3; range, 65-87). During the first week of 17beta-estradiol treatment, psychiatric symptoms diminished significantly (BPRS score decreased to a mean of 18.8, p < .001). Until the end of the second week of treatment, serum estradiol concentrations rose to near the values normally found during the follicular phase, and the patients became almost free of psychiatric symptoms. CONCLUSION: The reversal of psychiatric symptoms in all patients by treating documented estradiol deficiency suggests that estradiol plays a role in the pathophysiology and may have a role in the treatment of this condition. There was a rebound of psychotic symptoms in the 1 patient who discontinued estradiol treatment. Given the small number of patients, this area deserves further study. PMID- 10817100 TI - Serum valproate levels in 6 breastfeeding mother-infant pairs. AB - BACKGROUND: Women with bipolar disorder are at high risk for recurrence of an affective episode in the postpartum period, and treatment with a mood stabilizer may be indicated. Few data are available to inform the risk-benefit decision regarding the use of valproate for women with bipolar disorder who elect to breast-feed. METHOD: Serum valproate levels were obtained from 6 breastfeeding mother-infant pairs. All mothers had a diagnosis of bipolar disorder (Research Diagnostic Criteria) and were taking divalproex sodium as prophylaxis for or treatment of a recurrent affective episode. None of the mothers received valproate during pregnancy. RESULTS: The mothers had serum valproate levels near or within the therapeutic range (39.4 to 79.0 microg/mL). Infant serum levels were low, ranging from 0.7 to 1.5 microg/mL (0.9%-2.3% of maternal serum levels). No adverse clinical effects were observed in the infants. CONCLUSION: Serum valproate levels were low in nurslings of mothers treated with valproate. These data can be used to inform clinical decisions regarding the use of valproate during breastfeeding. PMID- 10817101 TI - Medication status and polycystic ovary syndrome in women with bipolar disorder: a preliminary report. AB - BACKGROUND: In patients with epilepsy, polycystic ovary (PCO) syndrome has been reported to be associated with the use of the anticonvulsant divalproex sodium. Whether PCO syndrome is associated with divalproex use in patients with bipolar disorder has not previously been explored. METHOD: Twenty-two female outpatients with a DSM-IV diagnosis of bipolar disorder who were between the ages of 18 and 45 years (inclusive) and who were taking lithium and/or divalproex (10, divalproex monotherapy; 10, lithium monotherapy; 2, divalproex/lithium combination therapy) were evaluated. Patients completed questionnaires about their medical, psychiatric, and reproductive health histories, and body mass indices were calculated. In the early follicular phase of their menstrual cycle, women were examined for hirsutism, given a pelvic ultrasound, and/or assessed for changes in laboratory values such as serum levels of testosterone, free testosterone, estradiol, estrone, dehydroepiandrosterone, dehydroepiandrosterone sulfate, luteinizing hormone, follicle-stimulating hormone, and 17-OH progesterone. RESULTS: All 10 patients on lithium monotherapy, 6 of 10 patients on divalproex monotherapy, and both of the patients on divalproex/lithium combination therapy reported some type of menstrual dysfunction, which, in 4 cases, had preceded the diagnosis of bipolar disorder. Hirsutism was not common in any group, but obesity was prominent in all groups. Ovarian ultrasound revealed an increased number of ovarian follicles in 1 patient taking lithium and in none of the patients taking divalproex. Hormonal screening did not indicate PCO-like changes in any patient. CONCLUSION: In this pilot study of bipolar patients, PCO-like changes were not seen in women receiving divalproex or lithium. However, independent of therapeutic agent used, the bipolar women in this study reported high rates of menstrual disturbances, suggesting that the hypothalamic-pituitary-gonadal axis may be compromised in some women with bipolar disorder. PMID- 10817102 TI - Prevalence of overweight and obesity in bipolar patients. AB - BACKGROUND: Patients who receive pharmacologic treatment for bipolar illness frequently gain weight. This study evaluated the prevalence of overweight and obesity in an unselected group of bipolar patients and matched reference subjects. METHOD: The prevalence of overweight, obesity, and central adiposity was evaluated in 89 euthymic bipolar (DSM-IV) patients and 445 reference subjects, matched for age and sex, using a cross-sectional study design. RESULTS: Female patients were more often overweight and obese than female reference subjects (chi2 = 9.18, df = 2, p = .01). The frequency of overweight was similar in male patients and male reference subjects, but male patients were more likely to be obese. Patients were more centrally obese than the general population in women (chi2 = 32.21, df = 1, p = <.001) and in men (chi2= 8.81, df = 1, p = .003). Patients treated with antipsychotic drugs were more obese than patients not receiving these drugs (chi2= 4.7, df = 1, p = .03). CONCLUSION: Body fat is more centrally distributed in pharmacologically treated bipolar patients than in matched population controls. Obesity is more prevalent in patients than in the general population. Obesity prevalence is clearly related to the administration of antipsychotic drugs. PMID- 10817103 TI - Clinical predictors of acute response with quetiapine in psychotic mood disorders. AB - BACKGROUND: In controlled studies of patients with schizophrenia, the atypical antipsychotic quetiapine, 300 mg/day, has been shown to be as effective in the treatment of positive and negative symptoms as haloperidol. However, little is known about the efficacy of quetiapine in patients with psychotic mood disorders. The purpose of this study was to assess the efficacy of quetiapine in the treatment of psychotic mood disorders in comparison with nonaffective psychotic disorders and identify clinical factors associated with quetiapine response. METHOD: In a naturalistic setting, by reviewing medical records, we assessed response to quetiapine and factors associated with response to quetiapine in 145 consecutive patients newly treated with the drug at a nonprofit academic psychiatric hospital. These patients had received a discharge diagnosis of bipolar disorder (manic, mixed, or depressive type), major depression with psychotic features, schizophrenia, schizoaffective disorder (bipolar or depressive type), delusional disorder, or psychosis not otherwise specified (NOS) according to DSM-IV criteria. RESULTS: Patients with a diagnosis of bipolar disorder, manic, mixed, or depressed and schizoaffective disorder, bipolar type displayed higher response rates (> 74%) compared with patients with schizophrenia. However, this finding did not achieve statistical significance. A diagnosis of major depression with psychotic features (p = .02) and longer duration of illness (p = .03) were associated with less chance of responding. CONCLUSION: Quetiapine may be a useful alternative or adjunctive treatment for patients with bipolar and schizoaffective disorders. PMID- 10817104 TI - The treatment of sedative-hypnotic dependence: evaluating clinical predictors of outcome. AB - BACKGROUND: The objectives of this 6-month prospective study were to evaluate the efficacy of detoxification treatment for sedative-hypnotic dependence, examine the demographic and clinical predictors of outcome, and determine whether anxiety or other psychiatric comorbidity has a negative impact on outcome. METHOD: Eighty two patients with alcohol or benzodiazepine dependence (DSM-IV diagnostic criteria) were consecutively recruited upon entering treatment and were assessed by clinical and semistructured interviews, the Global Assessment Scale, the Hamilton Rating Scale for Depression, the Beck Depression Inventory, the revised 90-item Symptom Checklist, and urine drug screening. RESULTS: Both alcohol- and benzodiazepine-dependent patients succeeded in reducing their reported use of sedative-hypnotic substances during the follow-up period. However, at 3 months, benzodiazepine-dependent patients fared less well than alcohol-dependent patients in terms of several outcome measures: they reported a lower rate of achieving abstinence, shorter periods of continuous abstinence, and more frequent drug use. At 6 months, the differences in outcome among the drug groups were not maintained. Variables such as sex, drug group, and indicators of psychiatric status had little impact on outcome measures. Benzodiazepine-dependent patients reported significant decreases in their level of anxiety over the follow-up period despite substantial reductions in benzodiazepine use. CONCLUSION: Clinicians may be encouraged regarding the detoxification of patients who have used benzodiazepines at high doses or for long periods of time, or who have comorbid anxiety or other psychiatric disorders. PMID- 10817105 TI - Bupropion sustained release versus paroxetine for the treatment of depression in the elderly. AB - BACKGROUND: Depression is a serious and widespread emotional disorder among the elderly. This study compared the efficacy and safety of bupropion sustained release (SR) with the selective serotonin reuptake inhibitor paroxetine in the treatment of major depression in elderly outpatients. METHOD: Elderly (> or = 60 years) outpatients with major depressive disorder (DSM-IV criteria) were evaluated in this 6-week multicenter, randomized, double-blind study comparing bupropion SR, 100-300 mg/day, and paroxetine, 10-40 mg/day. Efficacy was assessed by changes in scores on the Hamilton Rating Scales for Depression (HAM-D) and Anxiety (HAM-A) and the Clinical Global Impressions-Severity of Illness and Improvement scales. Safety was assessed by monitoring adverse events, vital signs, and body weight. RESULTS: A total of 100 patients ranging in age from 60 to 88 years were randomly assigned to treatment with bupropion SR (N = 48) or paroxetine (N = 52). Measurements of efficacy were similar between the 2 treatment groups, with both groups showing improved scores on all depression rating scales. Headache, insomnia, dry mouth, agitation, dizziness, and nausea occurred in > 10% of patients in both groups; somnolence, diarrhea, constipation, and anorexia occurred in > 10% of patients in the paroxetine group. No statistically significant differences between groups in vital signs or weight were found. CONCLUSION: Both bupropion SR and paroxetine were safe and effective for the treatment of depression in the elderly. Because of its favorable side effect profile, bupropion SR may provide a safe and effective nonserotonergic treatment alternative that is well suited as an antidepressant for the elderly. PMID- 10817106 TI - Nefazodone in patients with treatment-refractory posttraumatic stress disorder. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) is a highly prevalent and often chronic disorder among combat veterans, persisting in as many as 15% of Vietnam veterans for at least 20 years. Treatment response in veterans with combat related PTSD has been disappointing. Although anxiolytics, anticonvulsants, antipsychotics, and antidepressants have been tried, none has been consistently associated with improvement in all primary symptom domains (i.e., intrusive recollections, avoidance/numbing, and hyperarousal). This open-label study evaluated the use of nefazodone in a group of Vietnam veterans with chronic, treatment-refractory symptoms of PTSD. METHOD: Male outpatients with DSM-IV PTSD who had failed a minimum of 3 previous medication trials were eligible for the study. Nineteen Vietnam combat veterans entered the study and were treated with nefazodone, 100-600 mg/day, for 12 weeks. PTSD symptoms, anxiety, depression, sleep, sexual functioning, and adverse events were assessed weekly. RESULTS: Severity of depression lessened, as did PTSD symptoms of intrusive recollections, avoidance, and hyperarousal. Depressive symptom severity as measured by the Beck Depression Inventory decreased by a mean of 30%. Similarly, there was an overall drop in the intensity of PTSD symptoms as measured by the Clinician Administered PTSD Scale of 32% with a 26% improvement for symptoms of intrusion, 33% for avoidance, and 28% for arousal. In addition, improvements in sleep and sexual functioning were reported. The mean daily dose of nefazodone after 12 weeks was 430 mg (range, 200-600 mg/day). The most frequently reported side effects were headaches (53%), dry mouth (42%), and diarrhea (42%), but side effects tended to be mild and transient. CONCLUSION: In this group of Vietnam veterans with chronic treatment-refractory PTSD and multiple comorbid Axis I psychiatric disorders, nefazodone was well tolerated and effective. Larger, controlled studies are warranted. PMID- 10817107 TI - Risperidone in acutely exacerbated schizophrenia: dosing strategies and plasma levels. AB - BACKGROUND: The optimal risperidone dosing strategy for acute schizophrenia requires elucidation. Furthermore, plasma levels of risperidone and its active metabolite (9-hydroxyrisperidone) at a given dose vary greatly among different individuals. For patients who metabolize risperidone slowly, a medium dose results in excessively high plasma levels, which might be related to adverse events and perhaps poor response. We thus investigated whether dose reduction to diminish adverse reactions associated with ordinary risperidone doses could still yield efficacy for acutely exacerbated schizophrenia. METHOD: Thirty-one newly hospitalized Chinese patients with acute exacerbation of schizophrenia (DSM-IV) entered this prospective, 6-week open trial. Risperidone doses were titrated to 6 mg/day (if tolerable) over 3 days, but were lowered thereafter if side effects appeared. Efficacy and side effect assessments were conducted on days 0, 4, 14, 28, and 42. Endpoint steady-state plasma levels of risperidone and 9 hydroxyrisperidone were analyzed by high performance liquid chromatography with ultraviolet detection. RESULTS: Thirty patients completed the trial. Of them, 17 tolerated the 6-mg target dose well, while the other 13 received lower final doses (mean +/- SD = 3.6 +/- 0.9 mg, p = .0001) for curtailing treatment-emergent side effects. At endpoint, 92.3% of the 13 low-dose individuals responded to treatment (20% or more reduction in the total Positive and Negative Syndrome Scale score), compared with 52.9% of the 17 high-dose subjects (p < .05). No significant between-group differences were revealed in other minor efficacy measures. Of note, endpoint plasma levels of the active moiety (risperidone plus 9-hydroxyrisperidone) were similar between the low- and high-dose groups (40.4 +/ 31.1 ng/mL vs. 49.7 +/- 13.4 ng/mL, NS). CONCLUSION: The results of this preliminary trial suggest that up to 6 mg of risperidone is efficacious in treating patients with acute exacerbation of schizophrenia. Nearly 60% of the patients could tolerate a 6-mg dose. For the other 40%, reducing dosages to 3.6 +/- 0.9 mg for relieving side effects still yielded efficacy. The 2 dose groups were comparable in the endpoint steady-state plasma drug concentrations. PMID- 10817108 TI - Treatment of Kleine-Levin syndrome: melatonin on the starting block. PMID- 10817109 TI - Multiple rib fractures secondary to severe tardive dystonia and respiratory dyskinesia. PMID- 10817110 TI - Nefazodone-associated subjective complaints of burning sensations. PMID- 10817111 TI - Metrifonate for Alzheimer's disease patients. PMID- 10817112 TI - Urinary incontinence with risperidone. PMID- 10817113 TI - Underuse of antidepressants in major depression: prevalence and correlates in a national sample of young adults. AB - BACKGROUND: Epidemiologic studies have reported disturbingly low rates of treatment for major depression in the United States. To better understand this phenomenon, we studied the prevalence and predictors of antidepressant treatment in a national sample of individuals with major depression. METHOD: Between 1988 and 1994, 7589 individuals, aged 17-39 years and drawn from a national probability sample, were administered the Diagnostic Interview Schedule as part of the National Health and Nutrition Examination Survey. Interviewers asked about prescription drug use and checked medication bottles to record the name and type of medications. RESULTS: A total of 312 individuals, or 4.1% of the sample, met DSM-III criteria for current major depression. Only 7.4% of those with current major depression were being treated with an antidepressant. Among individuals with current major depression, being insured and having a primary care provider each predicted a 4-fold increase in odds of antidepressant treatment; telling the primary provider about depressive symptoms predicted a 10-fold increase in treatment. CONCLUSION: The study's findings support the notion that a serious gap exists between the established efficacy of antidepressant medications and rates of treatment for major depression in the "real world." Underreporting of depressive symptoms to providers and problems with access to general medical care appear to be 2 major contributors to this problem. PMID- 10817114 TI - Cancer prevalence estimates based on tumour registry data in the Surveillance, Epidemiology, and End Results (SEER) Program. AB - BACKGROUND: The Connecticut Tumor Registry (CTR) has collected cancer data for a sufficiently long period of time to capture essentially all prevalent cases of cancer, and to provide unbiased estimates of cancer prevalence. However, prevalence proportions estimated from Connecticut data may not be representative of the total US, particularly for racial/ethnic subgroups. The purpose of this study is to apply the modelling approach developed by Capocaccia and De Angelis to cancer data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute to obtain more representative US site specific cancer prevalence proportion estimates for white and black patients. METHODS: Incidence and relative survival were modelled and used to obtain estimated completeness indices of SEER prevalence proportions for all cancer sites combined, stomach, cervix uteri, skin melanomas, non-Hodgkin's lymphomas, lung and bronchus, colon/rectum, female breast, and prostate. For validation purposes, modelled completeness indices were computed for Connecticut and compared with empirical completeness indices (the ratio of Connecticut based prevalence proportion estimates using 1973-1993 data to 1940-1993 data). The SEER based modelled completeness indices were used to adjust SEER prevalence proportion estimates for white and black patients. RESULTS: Model validation showed that the adjusted SEER cancer prevalence proportions provided reasonably unbiased prevalence proportion estimates in general, although more complex modelling of the completeness indices is necessary for female cancers of the colon, melanoma, breast, cervix, and all cancers combined. The SEER-based cancer prevalence proportions are incomplete for most cancer sites, more so for women, whites, and at older ages. For all cancers combined, prevalence proportions tended to be higher for whites than blacks. For the site-specific cancers this was true for stomach, prostate, cervix uteri, and lung and bronchus (men only). For colon/rectal cancers the prevalence proportions were higher for blacks through ages 59 (men) and 64 (women), and then for the remaining ages they were higher for whites. Prevalence proportions were lowest for stomach cancer and highest for prostate and female breast cancers. Men experienced higher prevalence proportions than women for skin melanomas, non-Hodgkin's lymphomas, lung and bronchus, and colon/rectal cancers. CONCLUSION: The modelling approach applied to SEER data generally provided reasonable estimates of cancer prevalence. These estimates are useful because they are more representative of cancer prevalence than previously obtained and reported in the US. PMID- 10817115 TI - Anthropometric variables in relation to risk of breast cancer in middle-aged women. AB - BACKGROUND: Many epidemiological studies have assessed the relationships between anthropometric variables and breast cancer risk. However, methodological approaches for analysing these factors differ appreciably. Also, age when maximum height is achieved has been identified as a potential risk factor for breast cancer in premenopausal women, but this issue has not been studied in postmenopausal women. METHODS: The participants in this population-based case control study were postmenopausal women 50-64 years of age from the general female population of western Washington State. It included 479 women with incident primary breast cancer and 435 controls. RESULTS: This study found that: (i) women who gained over 70 pounds since age 18 had an increased risk of breast cancer relative to those who stayed within 10 pounds of their weight at age 18 (odds ratio [OR] = 2.7; 95% CI: 1.5-4.9), (ii) women with body mass indices (BMI) below what is considered healthy had a decreased risk (OR = 0.4; 95% CI: 0.2-1.1) while women with a BMI in the obese range had an increased risk of breast cancer (OR = 1.4; 95% CI: 1.0-2.1), and (iii) women who reached their maximum height at or after the age of 18 had a decreased risk of breast cancer compared to women who reached their maximum height at age 13 or younger (OR = 0.7; 95% CI: 0.5 1.0). CONCLUSIONS: By examining various anthropometric variables using clinically relevant strata, a clearer picture of how these variables relate to postmenopausal breast cancer risk was developed. Similar to younger women, postmenopausal women who reached their maximum height at later ages had a decreased risk of breast cancer. PMID- 10817116 TI - Variation in the interaction between familial and reproductive factors on the risk of breast cancer according to age, menopausal status, and degree of familiality. AB - BACKGROUND: Studies have found that reproductive factors might have a variable effect on the occurrence of breast cancer (BC) according to the existence or not of a family history of BC. The effect of a family history of BC on the risk of BC may also vary according to the age at diagnosis and the degree of kinship. This may confound the relation between familial risk and reproductive factors. A combined analysis was performed to study the interaction between familial risk and reproductive factors according to degree of familiality, age at interview and menopausal status. METHODS: The present analysis included 2948 cases and 4170 controls in seven case-control studies from four countries. The combined relative risks were estimated using a Bayesian random-effects logistic regression model. RESULTS: The main effects of reproductive life factors on the risk of BC are in agreement with previous studies. Two-way interactions between subject's age or menopausal status and a family history of BC were not significant. Although the three-way interaction between age, familial risk and parity was not significant, familial risk seemed to be increased slightly for women with high parity compared with women with low parity in the older age group, and seemed to be slightly decreased for women with high parity compared with women with low parity in younger women. The subject's age also appeared to have an effect on the interaction between familial risk and the age at first childbirth (P = 0.1). CONCLUSIONS: A possible influence of reproductive and menstrual factors on familial risk of BC has been suggested previously and was also evident in the present study. Three-way interactions between age, family history and parity or age at first childbirth might exist and they merit further investigation. PMID- 10817117 TI - Lifetime environmental exposure to tobacco smoke and primary lung cancer of non smoking Taiwanese women. AB - BACKGROUND: For a female population with a high lung cancer mortality rate, such as Taiwanese women, who smoke relatively rarely, but live in an environment with high male smoking prevalence, the risk and population burden of lung cancer due to environmental tobacco smoke (ETS) are relatively important. METHODS: An age matched case-control study was designed to investigate the effects of cumulative environmental exposure to tobacco smoke during childhood and adult life on lung cancer risk among non-smoking women in Taiwan. Information on passive smoking from all possible sources and life periods were obtained from interviews with 268 and 445 lifetime non-smoking cases and controls. Conditional logistic regression and synergism 'S' index were applied to the data to assess the independent and joint effects of passive smoking in different life stages while controlling for possible confounding variables. RESULTS: Risks of contracting lung cancer among women near-distantly exposed to the highest level of ETS in childhood (>20 smoker years) and in adult life (>40 smoker-years) were 1.8-fold (95% CI: 1.2-2.9) and 2.2-fold (95% CI: 1.4-3.7) higher than that among women being never exposed to ETS, and the two variables accounted for about 37% of tumours in this non-smoking female population. Children were found to be more susceptible to ETS than adults and such early exposure was found to modify the effect of subsequent tobacco smoke exposure in adult life based on an additive interaction model. CONCLUSIONS: Environmental tobacco smoke exposure occurring in childhood potentiates the effect of high doses of exposure in adult life in determining the development of lung cancer. Smoking prohibition would be expected to protect about 37% of non smoking Taiwanese women against lung cancer. PMID- 10817118 TI - Prospective study on the relation of cigarette smoking with cancer of the liver and stomach in an endemic region. AB - BACKGROUND: Smoking has not been confirmed as a risk factor for cancers of the liver and stomach. The authors examined prospectively the relationship between smoking and these cancers in an endemic region. METHODS: The data used were a cohort study on the relationship between lifestyle and health in the region having the highest liver cancer mortality in Japan. Of the cohort members, 4050 males aged > or =40 years were included in the present analysis with a 9-year mean follow-up. Cox proportional hazards regression was used to estimate relative risks (RR) for cancer of the liver, stomach, smoking-related sites and others, while adjusting for age, residence, and alcohol intake. RESULTS: By the end of the study period, 59 cases of liver cancer and 53 cases of stomach cancer were identified. Current smokers, compared to subjects who had never smoked, had a threefold risk of liver cancer (RR = 3.3; 95% CI: 1.2-9.5) and a twofold risk of stomach cancer (RR = 2.2; 95% CI: 0.8-5.7). Sub-cohort analysis showed that adjustment for history of chronic liver disease did not attenuate the risk of liver cancer. Light/medium smokers had almost the same risk of these cancers as heavy smokers, while they showed a relatively low risk of smoke-related cancers. CONCLUSIONS: The present results indicate that smoking is a risk factor of liver and stomach cancer in a population with a high background risk for these cancers. However, causal inferences should be made cautiously due to a lack of information on known risk factors. PMID- 10817119 TI - Occupational risk factors for urothelial carcinoma: agent-specific results from a case-control study in Germany. MURC Study Group. Multicenter Urothelial and Renal Cancer. AB - BACKGROUND: This multicentre population-based case-control study was conducted to estimate the urothelial cancer risk for occupational exposure to aromatic amines, polycyclic aromatic hydrocarbons (PAH), and chlorinated hydrocarbons besides other suspected risk factors. METHODS: In a population-based multicentre study, 1035 incident urothelial cancer cases and 4298 controls matched for region, sex, and age were interviewed between 1991 and 1995 for their occupational history and lifestyle habits. Exposure to the agents under study was self-assessed as well as expert-rated with two job-exposure matrices and a job task-exposure matrix. Conditional logistic regression was used to calculate smoking adjusted odds ratios (OR) and to control for study centre and age. RESULTS: Urothelial cancer risk following exposure to aromatic amines was only slightly elevated. Among males, substantial exposures to PAH as well as to chlorinated solvents and their corresponding occupational settings were associated with significantly elevated risks after adjustment for smoking (PAH exposure, assessed with a job-exposure matrix: OR = 1.6, 95% CI: 1.1-2.3, exposure to chlorinated solvents, assessed with a job task-exposure matrix: OR = 1.8, 95% CI: 1.2-2.6). Metal degreasing showed an elevated urothelial cancer risk among males (OR = 2.3, 95% CI: 1.4 3.8). In females also, exposure to chlorinated solvents indicated a urothelial cancer risk. Because of small numbers the risk evaluation for females should be treated with caution. CONCLUSIONS: Occupational exposure to aromatic amines could not be shown to be as strong a risk factor for urothelial carcinomas as in the past. A possible explanation for this finding is the reduction in exposure over the last 50 years. Our results strengthen the evidence that PAH may have a carcinogenic potential for the urothelium. Furthermore, our results indicate a urothelial cancer risk for the use of chlorinated solvents. PMID- 10817120 TI - Nicotine dependence and its familial aggregation in Chinese. AB - BACKGROUND: Nicotine dependence is a significant public health problem. This study attempted to characterize the prevalence and familial aggregation of nicotine dependence in China. METHODS: In 1998, we initiated a community-based cross-sectional study among residents of the Yuexi County in Anqing, China. A total of 991 current smokers from 488 randomly selected nuclear families were recruited and surveyed by use of the standardized Fagerstrom Test of Nicotine Dependence (FTND) questionnaire and the Revised Tolerance Questionnaire (RTQ). All study subjects were offspring in their respective nuclear families, and 478 male-male pairs (aged 41.7 +/- 12.1 years) were finally used for data analyses, because the number of female current smokers (n = 5) was too small. RESULTS: The correlation coefficient of the FTND and the RTQ scores was as high as 0.84 (P < 0.0001). Nicotine dependence, defined as an FTND score > or =8 or an RTQ score > or =28, had a prevalence of 12.7% and 11.1%, respectively. The respective sibling recurrent risk was 1.7 and 2.4, according to the FTND or the RTQ criteria. The adjusted odds ratios and 95% CI of nicotine dependence of second siblings in families in which the first sibling was nicotine dependent were 2.13 (95% CI: 1.02-4.43) and 3.50 (95% CI: 1.65-7.36) respectively, according to the FTND and RTQ criteria. CONCLUSIONS: The prevalence of nicotine dependence in male current smokers in China was comparable to that reported in previous US and European studies. Our findings suggest that genetic influences may play an important role in vulnerability to nicotine addiction. PMID- 10817121 TI - Tobacco smoking and risk of hip fracture in men and women. AB - BACKGROUND: Previous findings suggest that tobacco smoking increases the risk of hip fracture in women. A similar adverse effect of smoking is suspected to be present in men, but bone mineral density studies have raised the concern that men may be more sensitive to the deleterious effect of smoking on bone than women. In this study we prospectively determined the influence of current, previous, and cumulative smoking history on risk of hip fracture in men and women and addressed the issue of possible gender difference in the susceptibility to tobacco smoking. METHODS: Pooled data from three population studies conducted in Copenhagen with detailed information on smoking habit. A total of 13,393 women and 17,379 men, initially examined between 1964 and 1992, were followed until 1997 for first admission due to hip fracture. The relative risks (RR) of hip fracture associated with smoking were estimated by means of multiplicative Poisson regression models. RESULTS: During follow-up, 722 hip fractures were identified in women, and 447 in men. After adjustment for potential confounders, including body mass index, female current smokers had an RR of hip fracture of 1.36 (95% CI: 1.12-1.65) and male smokers 1.59 (95% CI: 1.04-2.43) relative to never smokers. In both sexes, the RR of hip fracture gradually increased by current and accumulated tobacco consumption. The RR were consistently higher in men than in women, but the test for interaction between sex and tobacco smoking was insignificant. After 5 years, male ex-smokers had an adjusted RR of 0.73 (95% CI: 0.55-0.98) relative to current smokers, while no significant decrease in risk was observed in female ex smokers (RR = 0.91; 95% CI: 0.72-1.17)). Approximately 19% of all hip fractures in the present study population were attributable to tobacco smoking. CONCLUSION: Tobacco smoking is an independent risk factor for hip fracture in men and women, and there appears to be no gender differences in smoking related risk. Smoking cessation reduces the risk of hip fracture in men after 5 years, while the deleterious effect of smoking seems to be more long-lasting in female ex-smokers. PMID- 10817122 TI - Saturated fat, vitamin C and smoking predict long-term population all-cause mortality rates in the Seven Countries Study. AB - BACKGROUND: The Seven Countries Study has shown that population mortality rates for various chronic diseases are related to diet and smoking. This paper addresses the associations between diet, smoking and 25-year all-cause mortality. METHODS: Baseline surveys were carried out between 1958 and 1964 on 12,763 middle aged men constituting 16 cohorts in seven countries. In 1987/88 equivalent food composites representing the average food intake of each cohort at baseline were collected and chemically analysed in one central laboratory. During 25 years of follow-up 5973 men died and age-adjusted population mortality rates were calculated for each cohort. RESULTS: Multivariate linear regression analyses showed that the population intake of saturated fat and the prevalence of smoking were positively associated with population all-cause mortality rates. Population vitamin C intake was inversely associated with all-cause mortality. It was calculated that a reduction in saturated fat intake of 5% of energy, a 20 mg/d increase in vitamin C and a 10% decrease in the prevalence of smokers may decrease the 25-year all-cause population mortality rate by 12.4% (95% CI: 5.6, 19.4%) at an average population all-cause mortality rate of 45%. CONCLUSION: At the population level saturated fat, vitamin C and cigarette smoking are important determinants of all-cause mortality. PMID- 10817123 TI - Association of thromboangiitis obliterans with cigarette and bidi smoking in Bangladesh: a case-control study. AB - BACKGROUND: In addition to cigarettes, bidi, made of unprocessed and low-grade tobacco, is being smoked widely in Bangladesh and in other south Asian countries. The cause-and-effect relationship is established between thromboangiitis obliterans (TAO) and smoking. However, type of smoking material(s) most strongly related to TAO is not yet determined. METHODS: We conducted a hospital-based case control study in Rajshahi, Bangladesh, to examine the relationship of type of smoking materials (cigarette versus bidi) with TAO on 103 pairs of cases and controls matched by age and sex during the period 1995 to 1996. The inclusion criteria for cases were newly diagnosed TAO and current smoker, while those for controls were current smokers admitted to the hospital due to non-cardiovascular diseases. RESULTS: Among the cases 35.0% and 65.0% were cigarette and bidi smokers, while among the controls 69.9% and 30.1%, respectively. Using logistic regression approach, considering cigarette smoking approximately 10 per day as reference, bidi smoking >20 per day (odds ratio [OR] = 34.76, 95% CI: 6.11 197.67) and 11-20 per day (OR = 7.12, 95% CI: 2.35-21.63) had greater risk of TAO after adjusting confounding factors. Respective OR for bidi smoking approximately 10 per day, cigarette smoking 11-20 per day and cigarette smoking >20 per day, were 2.18 (95% CI: 0.64-7.51), 3.81 (95% CI: 1.37-10.57) and 6.88 (95% CI: 1.87 25.30). CONCLUSION: Within the limits inherent to case-control study, our findings suggest that bidi smoking may well play a more important role in causing TAO than cigarettes. It leads to the speculation that unprocessed and low-grade tobacco used for producing bidi might play a more potent role to initiate TAO than cigarettes. PMID- 10817124 TI - Causality and temporality in the study of short-term effects of air pollution on health. AB - BACKGROUND: The relationship between air pollution and health is usually analysed by time series analysis. However, temporality (i.e. looking to see whether the cause precedes the effect) is rarely examined in detail. METHODS: Models were fitted to daily levels of black smoke, nitrogen dioxide, sulphur dioxide, ozone and mortality from Barcelona 1986-1995 to account for seasonality, environmental temperature, days of the week and influenza epidemics. Cross-correlations of the residuals were plotted for different lags. RESULTS: Clear evidence of a temporal relationship between mortality and air pollution found for all four pollutants in that changes in the pollutant preceded changes in mortality, implying causality. However the pattern of dependence was different for each pollutant. CONCLUSION: The cross-correlation plot is a useful tool in the analysis of air pollution time series. PMID- 10817125 TI - Seasonal variation in mortality in Scotland. AB - BACKGROUND: Seasonal patterns in mortality have been recognized for many years. This study assesses seasonal variation in mortality in Scotland between 1981 and 1993 and considers its association with socioeconomic status and outdoor temperature. METHODS: Lagged Poisson regression analysis of numbers of deaths and average weekly temperature with adjustment for serial autocorrelation and influenza epidemics. RESULTS: There was significant seasonal variation in weekly death rates with a difference of about 30% between a summer trough and a winter peak. This variation was principally attributable to respiratory disease, cerebrovascular disease and coronary artery disease. Seasonal variation in mortality fell from around 38% in 1981-1983 to around 26% in 1991-1993. There was no clear evidence of a relationship between socioeconomic status and seasonal mortality, however the extent of the fall in seasonal variation was greater in deprived areas than in affluent areas. Overall, a 1 degree C decrease in mean temperature was associated with a 1% increase in deaths one week later. The lag in this relationship varied by cause of death and underlying temperature. CONCLUSIONS: Seasonal variations in mortality and the relationship between temperature and mortality are a significant public health problem in Scotland. It is likely that the strength of this relationship is a result of the population being unable to protect themselves adequately from the effects of temperature rather than the effects of temperature itself. PMID- 10817126 TI - Chlamydia pneumoniae serological status is not associated with asthma in children or young adults. AB - BACKGROUND: The factors that cause the allergic sensitization and inflammation in asthma still remain to be clarified. A role for Chlamydia pneumoniae has been suggested although serological studies have produced conflicting findings. This study aims to clarify the relationship between asthmatic variables and C. pneumoniae serological status. METHODS: A case-control study was undertaken on an asthma-enriched subset from a longitudinal birth cohort. In all, 198 subjects (96 with self-reported asthma) had C. pneumoniae serology (microimmunofluorescence [MIF] IgG, IgA) undertaken at age 11 and age 21 and assessment made in relation to a number of asthma variables. RESULTS: The only statistically significant finding was in subjects self-reporting asthma at age 21 who had evidence of lower IgG titres (P = 0.046), a finding in the opposite direction to that expected from the hypothesis. Subjects with high IgG titres (> or =128) were less likely to have reported ever having asthma; odds ratio (OR) = 0.29, (95% CI: 0.10-0.87). No association existed between symptoms suggestive of asthma in the previous 12 months and either IgG (P = 0.127) or IgA (P = 0.189) antibody titres at age 21. Likewise, no association was found between symptoms suggestive of asthma in the previous two years and C. pneumoniae IgG antibody titre (P = 0.81) at age 11. There was no evidence of an association with any of the other variables examined at either age 11 or age 21. These included use of inhaled steroids, serum IgE levels, airway responsiveness, skin test evidence of atopy, or smoking status. CONCLUSION: The results of this study suggest that C. pneumoniae infection when diagnosed by MIF serology is not a major risk factor for the development of asthma in children and young adults. The study has not, however, addressed the role this organism may play in specific asthmatic subsets or asthma exacerbations. PMID- 10817127 TI - Primary and secondary infertility in sub-Saharan Africa. AB - BACKGROUND: No previous study has provided national estimates of the prevalence of primary and secondary infertility in sizeable areas of sub-Saharan Africa. METHODS: Primary infertility is measured by the proportion childless among women who entered their first marriage at least 7 years before date of censoring. Secondary infertility is measured by the 'subsequently infertile estimator' from parous ever-married women. Exposure begins at the age of the woman at the birth of her first child, and exposure ends when the woman is of an age, which is 5 years lower than her age at censoring. These last 5 years are used to determine her status as infertile or fertile at the last observation 5 years before censoring. A woman is considered infertile at last observation if she has had no livebirths during the last 5 years before censoring, otherwise she is considered fertile. A woman who has not given birth at age a or later is defined as being 'infertile subsequent to age a'. The index of the proportion subsequently infertile at age a is estimated as the number of women infertile subsequent to age a, divided by the total number of women observed at that age. Infertility is estimated for women age 20-44. RESULTS: Primary infertility is relatively low and it exceeds 3% in less than a third of the 28 African countries analysed. In contrast, elevated levels of secondary infertility prevail in most countries. Secondary infertility for women age 20-44 ranges from 5% in Togo to 23% in Central African Republic. CONCLUSIONS: It is feasible to gauge national levels of primary and secondary infertility from population based surveys including a birth history. The prevalence of infertility of pathological origin is so high in sub Saharan Africa that infertility is not merely an individual concern, it is a public health problem. PMID- 10817128 TI - Cerebral palsy and multiple births in China. AB - BACKGROUND: A population-based study on prevalence of cerebral palsy in multiple births has not been carried out in China. The purpose of this paper was to determine the prevalence of cerebral palsy in multiple births and to explore the influence of multiple pregnancy on cerebral palsy after controlling for birthweight. METHODS: A cross-sectional study of cerebral palsy was carried out among 388,192 children aged <7 years in seven cities of Jiangsu province in China. Information about birthweight and plurality was obtained from routine health care records. Pediatricians at city level diagnosed all cases. All the doctors involved had taken part in a training programme held by Beijing Medical University. Stratified analysis by birthweight and its standard normal deviate was employed to compare the prevalence of cerebral palsy in multiples and singletons. RESULTS: The prevalence of cerebral palsy for children aged <7 years in multiples was 9.7 per 1000 children (95% confidence interval [CI]: 6.5-14.0), which was 6.5 times that in singletons (95% CI: 4.4-9.3). The overall neonatal mortality rate was 60.9 per 1000 liveborn multiples, being highest (944.4 per 1000) in the 500-999 g birthweight groups. Most liveborn multiples weighing <1500 g at birth probably died from diseases related to very low birthweight prior to this study. The prevalence of cerebral palsy in multiple births was likely to be higher than that reported in developed countries for children weighing 1500-2499 g even though our data were from a cross-sectional study. When stratified by birthweight, the prevalence of cerebral palsy in multiples weighing <2500 g had tended to be lower than that of singletons in the same birthweight group. In contrast, in normal birthweight categories multiple births had a higher prevalence of cerebral palsy than singletons. When stratified by birthweight normal deviate, the prevalence of cerebral palsy in multiple births was uniformly higher than that in singletons in all birthweight strata and the prevalence of cerebral palsy among multiples appeared to be augmented as birthweight increased. CONCLUSION: The prevalence of cerebral palsy in multiples, 9.7 per 1000 children, is 6.5 times that in singletons. Survival of low birthweight infants is lower in China than in developed countries and survival quality of Chinese children weighing 1500-2499 g needs to be further improved. In terms of birthweight multiples and singletons may be heterogeneous. It might be difficult to directly use actual birthweight specific prevalence to compare the prevalence of cerebral palsy in multiples and singletons. Birthweight normal deviate specific prevalence of cerebral palsy suggests that multiple pregnancy is an independent risk factor for cerebral palsy in all birthweight groups. Multiples are in adverse circumstances very early in gestation and as the foetus matures the risk of cerebral palsy increases. PMID- 10817129 TI - Epidemiological models and related simulation results for understanding of contraceptive adoption in India. AB - BACKGROUND: For the first time, models using multilevel analysis of Indian data and related simulation results are reported. They take hierarchical structure into account and incorporate variables from all levels to get correct analysis and proper interpretation of data on current contraceptive use (including sterilization and modern methods). METHODS: The data from an Indian State, Uttar Pradesh (UP), collected by the National Family Health Survey (NFHS) conducted during 10 October 1992 to 22 February 1993 was used. For model I, 7851 currently married women who were neither pregnant nor had continuing post-partum amenorrhoea (PPA) were considered. For model II, these women with at least one child (n = 6748) were used. Two-level logistic regression analysis was carried out for which women's level (level 1) and PSU (Primary Sampling Unit) level (level 2) variables were considered. The results were considered significant at the 5% level of significance. Simulation analysis using each model was also carried out. RESULTS: Model I reveals that those more likely to adopt contraception were women exposed to a TV message (odds ratio [OR] = 1.3; 95% CI: 1.1-1.6); whose houses were pucca (bricks and mortar) (OR = 1.3; 95% CI: 1.1 1.5); who were educated to high school level and above (OR = 2.9; 95% CI: 2.2 3.7); whose husbands were literate with schooling of > or =11 years (OR = 1.7; 95% CI: 1.4-2.1); and who had > or =2 living sons (OR = 2.2; 95% CI: 1.1-4.4). Muslim and other religious women were less likely than Hindu women to adopt contraception (OR = 0.5; 95% CI: 0.4-0.6). Also, the PSU level availability of all weather road was positively associated with contraceptive adoption (OR = 1.4; 95% CI: 1.1-1.7). The PSU level variance, which is the unexplained PSU level variation after controlling for the considered characteristics, was significantly higher. The simulation results revealed that public health education (a TV message) was found to be more effective among less educated women. The PSU level availability of all weather road was as effective as public health education. Similar results were evident from the analysis of second data set (model II) with the noticeable finding that those whose last child is surviving are most likely to adopt contraception (OR = 8.82; 95% CI: 1.01-77.38). CONCLUSIONS: These results reveal that the survival status of the last child has a marked effect on the adoption of contraception in UP. They further support the idea that public health education (a TV message) is more effective among less educated women. Also, the PSU level presence of all weather road is equally effective. Consideration of higher level variables provides not only more accurate results but also important public health clues to help the policy planners. PMID- 10817130 TI - Physical activity and hip fracture: a population-based case-control study. Swedish Hip Fracture Study Group. AB - BACKGROUND: A growing body of literature suggests that physical activity may be a protective factor against hip fracture. METHODS: To study the association between hip fracture risk and recreational physical activity at various ages, changes in activity during adult life, occupational physical activity and how risks vary by adult weight change, we performed a population-based case-control study among postmenopausal women aged 50-81 years residing in six counties in Sweden in 1993 1995. The analysis consisted of 1327 women with hip fracture and 3262 randomly selected controls. Information on leisure physical activity before age 18, at 18 30 years and during recent years was based on a questionnaire. Data on occupational physical activity were collected through an independent classification of job titles obtained from record linkage with census data from 1960, 1970 and 1980. RESULTS: There was a protective effect of recent leisure physical activity. Compared to women who reported no leisure activity, the odds ratios (OR) were 0.79 (95% CI: 0.62-1.00), 0.67 (95% CI: 0.54-0.84) and 0.48 (95% CI: 0.39-0.60) for women who exercised <1 h per week, 1-2 h per week, and 3+ h per week, respectively. These decreased OR were more pronounced in women who had lost weight after 18 years of age than in those who had gained weight. Women with high physical activity at both 18-30 years and during recent years did not have a stronger protection than those with isolated high activity late in life, after accounting for recent activity. Occupational physical activity was not associated with hip fracture risk in this study. CONCLUSIONS: Recent physical activity is protective against hip fracture. The protective effect is most pronounced in women who had lost weight after age 18. PMID- 10817131 TI - Risks older drivers face themselves and threats they pose to other road users. AB - BACKGROUND: Although there is an ever increasing literature on older drivers, there is no comprehensive up-to-date presentation of how older drivers are impacted by traffic safety, and how they impact the road safety of others. METHODS: This paper uses 1994-1996 US data to determine how many rates related to traffic safety depend on the age and sex of road users (fatalities, fatalities per licensed driver, etc.) Threats drivers pose to other road users are estimated by driver involvement in pedestrian fatality crashes. RESULTS: It is found that renewing the licence of a 70-year-old male driver for another year poses, on average, 40% less threat to other road users than renewing the license of a 40 year-old male driver. The fatality risks drivers themselves face generally increase as they age, with the increased risk of death in the same severity crash being a major contributor. If this factor is removed, crash risks for 70-year-old male drivers are not materially higher than for 40-year-old male drivers; for female drivers they are. CONCLUSIONS: Most driver rates increase substantially by age 80, in many cases to values higher than those for 20-year-olds. Given that a death occurs, the probability that it is a traffic fatality declines steeply with age, from well over 20% for late teens through mid twenties, to under one per cent at age 65, and under half a per cent at age 80. PMID- 10817132 TI - Parkinson's disease mortality and pesticide exposure in California 1984-1994. AB - BACKGROUND: In the last two decades reports from different countries emerged associating pesticide and herbicide use with Parkinson's disease (PD). California growers use approximately 250 million pounds of pesticides annually, about a quarter of all pesticides used in the US. METHODS: We employed a proportional odds mortality design to compare all cases of PD recorded as underlying (1984 1994) or associated causes (1984-1993) of death occurring in California with all deaths from ischaemic heart disease (ICD-9 410-414) during the same period. Based on pesticide use report data we classified California counties into several pesticide use categories. Agricultural census data allowed us to create measures of percentage of land per county treated with pesticides. Employing logistic regression models we estimated the effect of pesticide use controlling for age, gender, race, birthplace, year of deaths, and education. RESULTS: Mortality from PD as the underlying cause of death was higher in agricultural pesticide-use counties than in non-use counties. A dose response was observed for insecticide use per county land treated when using 1982 agricultural census data, but not for amounts of restricted pesticides used or length of residency in a country prior to death. CONCLUSIONS: Our data show an increased PD mortality in California counties using agricultural pesticides. Unless all of our measures of county pesticide use are surrogates for other risk factors more prevalent in pesticide use counties, it seems important to target this prevalent exposure in rural California in future studies that use improved case finding mechanisms and collect pesticide exposure data for individuals. PMID- 10817133 TI - Estimation of the prevalence of epilepsy in the Benin region of Zinvie using the capture-recapture method. AB - BACKGROUND: The prevalence of epilepsy was estimated in two villages of 3134 inhabitants, in Benin, in April and May 1997 using the capture-recapture method. METHODS: Information was obtained from (i) a door-to-door cross-sectional study, (ii) a non-medical source consisting of key informants (traditional practitioners, teachers, village leaders, and religious representatives) and (iii) a medical source through evaluation of medical records in health centres. In all the three situations, the diagnosis of epilepsy was confirmed by a neurologist. RESULTS: The door-to-door survey found 50 epileptics, i.e. a prevalence of 15.9 per 1000. The non-medical source found 26 patients. The medical source found only four patients. In total, 66 epileptics were found by combining the three sources, giving a prevalence of 21.1 per 1000. After application of the capture-recapture method, the estimated number of cases from the door-to-door survey and non-medical source was 105, and 110 cases when the medical source was considered as well. The respective prevalences were 33.5 per 1000, and 35.1 per 1000. CONCLUSIONS: The door-to-door survey has been usefully improved by using key informants. The epilepsy prevalence estimate found by capture-recapture is clearly higher than that found by traditional cross sectional methods, and could better depict the frequency of epilepsy in Africa. PMID- 10817134 TI - Accuracy of cause-of-death coding in Taiwan: types of miscoding and effects on mortality statistics. AB - BACKGROUND: The objectives of this study were to assess the accuracy of cause-of death coding, determine the extent to which coders follow the selection rules of coding set out in the International Classification of Diseases, 9th Revision (ICD 9), and the effects of miscoding on mortality statistics in Taiwan. METHOD: A systematic sample of 5621 death certificates was reviewed. The underlying cause of death (UCD) selected by the reviewer for each death certificate was compared with that selected by the original coder. The UCD was selected according to ACME (Automated Classification of Medical Entities) Decision Tables. RESULTS: The overall agreement rates between the reviewer and coders according to the three digit and two-digit categories of ICD-9 were 80.9% and 83.9%, respectively. Good agreement was found for malignant neoplasms (kappa = 0.94) and injuries and poisoning (kappa = 0.97), but there was poor agreement for nephrotic diseases (kappa = 0.74), hypertension-related diseases (kappa = 0.74), and cerebral infarction (kappa = 0.77). Reasons for disagreements included disagreement in nomenclature (42.8%), inappropriate judgement of causal relationships (41.5%), and incorrect interpretation of Selection Rule 3 and Modification Rules (15.7%). CONCLUSION: This study showed various levels of agreement for different diseases between the reviewer and the original coders in selection of the UCD. Owing to the 'compensatory effect of errors', the national mortality statistics were not affected significantly. The national administration should undertake routine internal studies to control the quality of UCD coding practices. PMID- 10817135 TI - Behavioural and serological human immunodeficiency virus risk factors among female commercial sex workers in Cambodia. AB - BACKGROUND: The spread of human immunodeficiency virus (HIV) in Cambodia is mainly caused by sexual transmission and the high-risk group in this country are female commercial sex workers (CSW). There are two types of CSW, direct CSW (DCSW) and indirect CSW (IDCSW), who are different from each other in sexual activities. This study was conducted in order to describe the risk factors on HIV for each type of CSW, and to establish effective preventive strategies against the HIV epidemic among CSW. METHODS: The participants, 143 DCSW and 94 IDCSW, were interviewed using a questionnaire to determine their demographic characteristics and behaviour. Blood samples were taken for serological tests on HIV, Chlamydia trachomatis and syphilis. The association between their behavioural pattern and their serological results was analysed. RESULTS: The questionnaire study showed that IDCSW had a riskier behavioural pattern than DCSW. The HIV seroprevalence rates of the DCSW and the IDCSW were 52.4% and 22.3%, respectively. Univariate logistic analyses showed a significant association between HIV antibody (HIV-Ab) and current age, age at commencement of commercial sex work, duration of commercial sex work, and the seropositivity of Chlamydia trachomatis-IgG antibody (CT-IgG-Ab) among the DCSW. The analyses also showed a significant relationship between HIV-Ab and CT-IgG-Ab among the IDCSW. CONCLUSIONS: Improving condom use rate is very important in order to prevent an HIV epidemic among the two types of CSW. This study also suggests it is important to prevent sexually transmitted disease (STD) such as Chlamydia trachomatis infection. The STD control programme could be efficient for HIV prevention, especially among DCSW. PMID- 10817136 TI - A spatial statistical approach to malaria mapping. AB - BACKGROUND: Good maps of malaria risk have long been recognized as an important tool for malaria control. The production of such maps relies on modelling to predict the risk for most of the map, with actual observations of malaria prevalence usually only known at a limited number of specific locations. Estimation is complicated by the fact that there is often local variation of risk that cannot be accounted for by the known covariates and because data points of measured malaria prevalence are not evenly or randomly spread across the area to be mapped. METHOD: We describe, by way of an example, a simple two-stage procedure for producing maps of predicted risk: we use logistic regression modelling to determine approximate risk on a larger scale and we employ geo statistical ('kriging') approaches to improve prediction at a local level. Malaria prevalence in children under 10 was modelled using climatic, population and topographic variables as potential predictors. After the regression analysis, spatial dependence of the model residuals was investigated. Kriging on the residuals was used to model local variation in malaria risk over and above that which is predicted by the regression model. RESULTS: The method is illustrated by a map showing the improvement of risk prediction brought about by the second stage. The advantages and shortcomings of this approach are discussed in the context of the need for further development of methodology and software. PMID- 10817137 TI - Ten years of serological surveillance in England and Wales: methods, results, implications and action. AB - BACKGROUND: The first age-stratified serological survey of antibody to measles, mumps and rubella in the UK was conducted in 1986/87 prior to the introduction of MMR vaccine into the immunization programme. Serum collection and testing have continued annually, allowing trends over time to be monitored. These sera have also been available for ad hoc surveys of other infections. METHODS: Residual sera are collected in participating laboratories and sent to a central store where they are irrevocably unlinked from identifying data. A unique identity number is assigned to each serum and details of age and sex are collated on a database. The sera are accessed for testing as required. RESULTS: The results of recurring and other surveys performed over the last ten years are presented. These demonstrate that opportunistic serum samples are an ideal resource for serological surveillance programmes. CONCLUSIONS: The serological surveillance programme has provided past exposure profiles for many infections. These data have resulted in a number of national policy changes and have been instrumental in shaping the UK vaccination programme. PMID- 10817138 TI - Underestimation of Mycobacterium tuberculosis infection in HIV-infected subjects using reactivity to tuberculin and anergy panel. AB - BACKGROUND: This study aimed to evaluate purified protein derivative (PPD) reactivity and its interrelationship with anergy panel and CD4+ lymphocytes in HIV-infected subjects as compared to PPD reactivity in HIV-uninfected individuals in a tuberculosis endemic and high Bacillus Calmette-Guerin (BCG) coverage environment. METHODS: Clients of four Mexico City HIV detection centres were screened for HIV-1 antibodies (ELISA or haemagglutination, Western Blot); reactivity to PPD (Mantoux PPD, 5TU RT-23), Candida (1:1000, 0.1 ml), and tetanus toxoid (10Lf, 0.1 ml); and CD4+ T cells. Active tuberculosis was excluded. Informed consent was obtained. RESULTS: From 5130 clients 1168 subjects were enrolled; of these 801 (68.6%) were HIV positive. Reactivity to PPD among HIV positive subjects was found in 174 (22%), 261 (32.6%), and 296 (37%), at PPD cutoff levels of > or =10 mm, > or =5 mm, and > or =2 mm as compared to 224 (61%) of 367 HIV-negative individuals' reactors to PPD (> or =10 mm) (P < 0.001). After exclusion of anergic individuals using two cutoff levels for cutaneous allergens (< or =2 mm and < or =5 mm), PPD reactivity between HIV-infected and uninfected individuals continued to be significantly different. Only HIV-infected individuals with CD4+ T cells > or =500 cells/mm3 had similar reactivity to PPD as HIV-uninfected individuals. Variables associated with PPD reactivity were CD4+ T cell counts, BCG scar, HIV infection and age. CONCLUSIONS: PPD reactivity was useful to diagnose tuberculosis infection only among HIV-infected individuals with CD4+ counts > or =500 cells/mm3. Among individuals with lower counts, lowering cutoff levels or using anergy panel did not permit comparable reactivity as that observed among HIV-uninfected individuals. PMID- 10817139 TI - Serological analysis of a cryptosporidiosis epidemic. AB - BACKGROUND: A cryptosporidiosis epidemic occurred among residents and visitors to Collingwood, Ontario, during March 1996. Fifty-five per cent of 36 confirmed cases were Collingwood visitors and 57% of Collingwood resident cases were under 10 years of age. The low level of reported diarrhoeal illness among adult Collingwood residents caused government officials and physicians to question whether an epidemic had occurred in Collingwood. METHODS: To better evaluate the extent of the epidemic, anonymous surplus sera from 89 adult Collingwood residents, collected for routine tests prior to, during and after the epidemic, and from 80 adult Toronto residents were tested using a Western blot assay for IgG antibody response to two Cryptosporidium antigen groups (15/17-kDa and 27 kDa). RESULTS: For sera collected from 1 January 1996 to 17 June 1996, a higher fraction of Collingwood residents had a detectable serological response (P < 0.002) and the mean intensity of serological responses was higher for Collingwood than Toronto residents (P < 0.001). The mean intensity of serological responses for Collingwood residents was higher in specimens drawn during the 8 weeks following the initial case reports compared to those drawn before or after this period (15/17-kDa, P < 0.02; 27-kDa, P < 0.10). CONCLUSIONS: These elevated serological responses indicate that Cryptosporidium infections among Collingwood residents likely occurred more commonly than illness reports suggested, consistent with a community-wide cryptosporidiosis epidemic. Similar studies should be considered in future suspected cryptosporidiosis epidemic investigations. PMID- 10817140 TI - Reduction in male births among workers exposed to metal fumes. PMID- 10817141 TI - What are the major correlates of macronutrient selection in Western populations? AB - In order to better understand the factors that may influence and regulate the intake of the macronutrients carbohydrate, fat and protein a 7 d diet diary technique was employed to study eating behaviour in the natural environment of free-living human subjects. In general, factors that promote energy intake tend to promote fat and protein intake to a greater extent than carbohydrate intake. This increased intake occurs as the result of: environmental factors such as social facilitation and the time of day, week or lunar phase; subjective factors such as hunger and elation; individual difference factors such as obesity, restraint and ageing. There are indications that the intake of macronutrients is regulated by negative feedback systems. In the short term, the amount of protein remaining in the stomach at the onset of a meal appears to have a restraining effect on intake, especially protein intake. Over several days, macronutrient intake appears to be affected by a nutrient-specific delayed negative feedback. Protein intake during 1 d is negatively associated with protein intake 2 and 3 d later, while carbohydrate intake is negatively related to later carbohydrate intake, and fat intake to later fat intake; both peaking after a 2 d delay. Studies of the intakes of twins suggested that many aspects of the control of macronutrient intake are influenced by inheritance; these factors include the overall amounts ingested, the before-meal stomach contents and the responsiveness of the subject to the negative impact of the stomach contents. The results indicate that macronutrient intakes are regulated by multiple persistent processes that are to a large extent inherited. PMID- 10817142 TI - Diet selection and animal state: an integrative framework. AB - In the present paper we deal with the problems of explaining and predicting diet selection of animals under controlled conditions, i.e. conditions that can be described and in which any influences of the environment can be either controlled or at least monitored. Diet selection is considered within an integrative framework of feeding behaviour that views both food intake and diet selection as an outcome of the animal's internal state and knowledge of the feeding environment. Three questions that arise from the framework are considered: (1) how do animals learn about foods available to them as a choice? (2) what changes in internal state affect diet selection? (3) how much time is needed for a change in the animal's internal state to be detected and for it to react to this change through a modification of its diet selection? It is proposed that animals have developed behavioural mechanisms that allow them to recognize foods on the basis of their nutritional as well as other properties. The rate at which animals learn about foods depends largely on the extent of the animal's deficiency and on the extent of the post-ingestive consequences induced by the foods. There is little evidence that animals modify their diet selection in response to short-term systemic fluctuation of their internal environment. On the other hand, long-term changes in the internal state of the animal lead to consequent long-term changes in diet selection. The time needed for a change in diet selection to be observed depends on the deviation created in the animal's internal state, either as a result of a physiological change or as a consequence of feeding. Thus, a more appropriate question to consider is not 'what time period matters to the animal?' but 'how much change or deviation in the internal state is the animal prepared to accept?' PMID- 10817143 TI - High-fat and low-fat (behavioural) phenotypes: biology or environment? AB - It is now widely accepted that obesity develops by way of genetic mechanisms conferring specific dispositions which interact with strong environmental pressures. It is also accepted that certain dispositions constitute metabolic risk factors for weight gain. It is less well accepted that certain patterns of behaviour (arising from biological demands or environmental influences) put individuals at risk of developing a positive energy balance and weight gain (behavioural risk factors). Relevant patterns of behaviour include long-lasting habits for selecting and eating particular types of foods. Such habits define two distinct groups characterized as high-fat (HF) and low-fat (LF) phenotypes. These habits are important because of the attention given to dietary macronutrients in body-weight gain and the worldwide epidemic of obesity. Considerable evidence indicates that the total amount of dietary fat consumed remains the most potent food-related risk factor for weight gain. However, although habitual intake of a high-fat diet is a behavioural risk factor for obesity, it does not constitute a biological inevitability. A habitual low-fat diet does seem to protect against the development of obesity, but a high-fat diet does not guarantee that an individual will be obese. Although obesity is much more prevalent among HF than LF, some HF are lean with BMI well within the normal range. The concept of 'different routes to obesity' through a variety of nutritional scenarios can be envisaged, with predisposed individuals varying in their susceptibility to different dietary inputs. In a particular subgroup of individuals (young adult males) HF and LF displayed quite different profiles of appetite control, response to nutrient challenges and physiological measures, including BMR, RQ, heart rate, plasma leptin levels and thermogenic responses to fat and carbohydrate meals. These striking differences suggest that HF and LF can be used as a conceptual tool to investigate the relationship between biology and the environment (diet) in the control of body weight. PMID- 10817144 TI - Assuaging nutritional complexity: a geometrical approach. AB - We have introduced a framework that enables the identification of the important elements in complex nutritional systems, and the quantification of the interactions among them. These interactions include those among the multiple constituents of the ingesta, as well as between behavioural (ingestive) and physiological (post-ingestive) components of nutritional homeostasis. The resulting descriptions provide a powerful means to generate and test hypotheses concerning the mechanisms, ecology and evolution of nutritional systems. We provide an overview of the key concepts involved in our scheme, and then introduce four examples in which the framework is used to develop and test hypotheses. In the first example we use comparative methods based on a data set of 117 insect species to test a prediction about the relationship between evolving an association with bacterial endosymbionts and the composition of the optimal diet. Second, using two species of locusts (a grass specialist and a generalist), we consider the relationship between an animal's diet breadth and the decision rules employed when feeding on foods containing suboptimal protein: carbohydrate values. Third, we introduce a mathematical model that predicts the dose-response properties of gustatory systems in the context of nutritional homeostasis. Finally, we consider the interaction between tannic acid and macronutrient balance in the diet of locusts. PMID- 10817145 TI - Effects of extreme environments on food intake in human subjects. AB - Effects of extreme environments on food intake in human subjects are analysed as behavioural and physiological adaptations to annual and circadian rhythms, temperature and altitude. Effects of the environment on food intake through food availability have direct consequences on energy balance and body weight. Different geographical regions show variations in the composition of dietary foods, i.e. the relative proportions of carbohydrate, protein and fat. In developing countries the annual cycle appears to affect body weight through dependence on food availability. In West-European countries this effect appears to depend on physical activity. Energy and macronutrient intakes appear to follow a circadian pattern, with breakfast being relatively high in carbohydrate and dinner being relatively high in fat. In cold conditions, maintaining an adequate food intake is important in sustaining normal physiological responses to cold. Evidence for a possible cold-induced increase in appetite is poor. A condition influencing level of intake is the palatability of the food. High altitude, i.e. hypobaric hypoxia, appears to reduce appetite, energy intake and body mass, irrespective of acute mountain sickness (AMS). Meal size is reduced and meal frequency increased. Under circumstances of AMS, dissociation between appetite and hunger occurs. Thus, spontaneous adaptation to extreme environments requiring increased energy intake occurs first by adaptation of body weight to a new energy balance. In general, prevention of a negative energy balance occurs by learning with respect to food intake. After return to normal, adjustment of energy intake to the original energy balance occurs with restoration of body weight. PMID- 10817146 TI - Habitat selection according to the ability of animals to eat, digest and detoxify foods. AB - Large herbivores play a major role in shaping vegetation community dynamics through selective consumption of particular plants and plant communities. An understanding of the factors influencing diet selection at the level of individual bites ('bite scale') is important for prediction of the impact of herbivores on vegetation at the habitat scale. Bite-scale diet selection represents an integration of the twin goals of maximizing nutrient intake and minimizing toxin intake. Recent research with ruminants in pen-fed situations has shown that animals are able to make choices between artificial foods that maximize growth and other production variables. The role of post-ingestive feedback as an important mechanism for allowing animals to assess the nutritional quality of particular foods, and so select optimal diets, has been recognized in a number of recent experiments. Our understanding of the role of toxin intake minimization in diet selection decisions is more rudimentary. An important advance in the last decade has been the acknowledgement of the role of post ingestive feedback and learning as a mechanism for avoidance of dietary toxicity. Further research is required to assess the importance of these processes in relation to free-grazing animals. The extent to which an understanding of bite scale diet selection can be used to predict habitat utilization is not well understood. At the habitat scale additional factors such as predator avoidance, social constraints, avoidance of parasitism and microclimatic effects have an important influence on foraging decisions. Future research needs to focus on developing a quantitative understanding of such decisions at the habitat scale. PMID- 10817147 TI - Social determinants of food choice. AB - Food choice is influenced by a large number of factors, including social and cultural factors. One method for trying to understand the impact of these factors is through the study of attitudes. Research is described which utilizes social psychological attitude models of attitude-behaviour relationships, in particular the Theory of Planned Behaviour. This approach has shown good prediction of behaviour, but there are a number of possible extensions to this basic model which might improve its utility. One such extension is the inclusion of measures of moral concern, which have been found to be important both for the choice of genetically-modified foods and also for foods to be eaten by others. It has been found to be difficult to effect dietary change, and there are a number of insights from social psychology which might address this difficulty. One is the phenomenon of optimistic bias, where individuals believe themselves to be at less risk from various hazards than the average person. This effect has been demonstrated for nutritional risks, and this might lead individuals to take less note of health education messages. Another concern is that individuals do not always have clear-cut attitudes, but rather can be ambivalent about food and about healthy eating. It is important, therefore, to have measures for this ambivalence, and an understanding of how it might impact on behaviour. PMID- 10817148 TI - Conditioned food aversions: principles and practices, with special reference to social facilitation. AB - Conditioned food aversion is a powerful experimental tool to modify animal diets. We have also investigated it as a potential management tool to prevent livestock from grazing poisonous plants such as tall larkspur (Delphinium barbeyi), white locoweed (Oxytropis sericea) and ponderosa pine (Pinus ponderosa) on western US rangelands. The following principles pertain to increasing the strength and longevity of aversions: mature animals retain aversions better than young animals; novelty of the plant is important, although aversions can be created to familiar plants; LiCl is the most effective emetic, and the optimum dose for cattle is 200 mg/kg body weight; averted animals should be grazed separately from non-averted animals to avoid the influence of social facilitation which can rapidly extinguish aversions. Social facilitation is the most important factor preventing widespread application of aversive conditioning. When averted animals see other animals eat the target food they will sample it, and if there is no adverse reaction they will continue eating and extinguish the aversion. However, if averted animals can be grazed separately, aversions will persist. Aversive conditioning may provide an effective management tool to prevent animals from eating palatable poisonous plants that cause major economic loss. PMID- 10817149 TI - Pre-operative nutritional assessment. AB - Protein-energy undernutrition, or the possibility of its development, has been documented to occur frequently in surgical patients admitted to hospital. Nutritional status is known to deteriorate over the course of the hospital stay, with poor awareness by medical and nursing staff as to the deleterious effects of impaired nutritional status on clinical outcome and hospital costs. While there is no consensus on the best method for assessment of the nutritional status of surgical patients pre-operatively, there are a number of techniques available. These techniques can be divided into two types, those suitable for screening for nutrition risk on admission to hospital and those used to fully assess nutritional status. Both techniques have their limitations, but if used correctly, and their limitations recognized, should identify the appropriate degree of nutritional intervention for an individual patient in a timely and cost effective manner. The techniques currently available for nutritional screening and nutritional assessment are reviewed, and their applicability to the Irish setting are discussed in the present paper. PMID- 10817150 TI - Immunonutrition and surgical practice. AB - Immunonutrition generally refers to the effect of the provision of specific nutrients on the immune system. These nutrients typically have immunoenhancing properties, and recent advances in nutrition support involve studies designed to exploit the desirable biological properties of these nutrients. The term immunonutrition strictly implies that we are focusing on the effect of certain nutrients on aspects of the immune system. However, in reality immunonutrition also refers to studies that not only examine the function of lymphocytes and leucocytes, but which also study the influence of key nutrients on the acute phase response, the inflammatory response and on gastrointestinal structure and function. The interest, therefore, is on the impact of immunonutrition on all aspects of host defence mechanisms in response to a catabolic stress. Major surgery evokes an acute-phase response, a transient immunosuppression and alterations in gastrointestinal function. Normal function is usually restored after a few days; however, in a subgroup of patients homeostasis may be lost and development of the systemic inflammatory response syndrome (SIRS) ensues. Results of recent clinical trials suggest that provision of immunomodulatory nutrients, including glutamine, arginine, n-3 polyunsaturated fatty acids and dietary nucleotides, may promote restoration of normal tissue function post-operatively and prevent the occurrence of SIRS. PMID- 10817151 TI - Observation of intramyocellular lipids by means of 1H magnetic resonance spectroscopy. AB - Magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) are being increasingly used for investigations of human muscle physiology. While MRI reveals the morphology of muscles in great detail (e.g. for the determination of muscle volumes), MRS provides information on the chemical composition of the tissue. Depending on the observed nucleus, MRS allows the monitoring of high energy phosphates (31P MRS), glycogen (13C MRS), or intramyocellular lipids (1H MRS), to give only a few examples. The observation of intramyocellular lipids (IMCL) by means of 1H MRS is non-invasive and, therefore, can be repeated many times and with a high temporal resolution. MRS has the potential to replace the biopsy for the monitoring of IMCL levels; however, the biopsy still has the advantage that other methods such as those used in molecular biology can be applied to the sample. The present study describes variations in the IMCL levels (expressed in mmol/kg wet weight and ml/100 ml) in three different muscles before and after (0, 1, 2, and 5 d) marathon runs for a well-trained individual who followed two different recovery protocols varying mainly in the diet. It was shown that the repletion of IMCL levels is strongly dependent on the diet post exercise. The monitoring of IMCL levels by means of 1H MRS is extremely promising, but several methodological limitations and pitfalls need to be considered, and these are addressed in the present review. PMID- 10817152 TI - NMR of glycogen in exercise. AB - Natural-abundance 13C NMR spectroscopy is a non-invasive technique that enables in vivo assessments of muscle and/or liver glycogen concentrations. Over the last several years, 13C NMR has been developed and used to obtain information about human glycogen metabolism with diet and exercise. Since NMR is non-invasive, more data points are available over a specified time course, dramatically improving the time resolution. This improved time resolution has enabled the documentation of subtleties of muscle glycogen re-synthesis following severe glycogen depletion that were not previously observed. Muscle and liver glycogen concentrations have been tracked in several different human populations under conditions that include: (1) muscle glycogen recovery from intense localized exercise with normal insulin and with insulin suppressed; (2) muscle glycogen recovery in an insulin resistant population; (3) muscle glycogen depletion during prolonged low intensity exercise; (4) effect of a mixed meal on postprandial muscle and liver glycogen synthesis. The present review focuses on basic 13C NMR and gives results from selected studies. PMID- 10817153 TI - Introduction to in vivo 31P magnetic resonance spectroscopy of (human) skeletal muscle. AB - 31P magnetic resonance spectroscopy (MRS) offers a unique non-invasive window on energy metabolism in skeletal muscle, with possibilities for longitudinal studies and of obtaining important bioenergetic data continuously and with sufficient time resolution during muscle exercise. The present paper provides an introductory overview of the current status of in vivo 31P MRS of skeletal muscle, focusing on human applications, but with some illustrative examples from studies on transgenic mice. Topics which are described in the present paper are the information content of the 31P magnetic resonance spectrum of skeletal muscle, some practical issues in the performance of this MRS methodology, related muscle biochemistry and the validity of interpreting results in terms of biochemical processes, the possibility of investigating reaction kinetics in vivo and some indications for fibre-type heterogeneity as seen in spectra obtained during exercise. PMID- 10817154 TI - Arterio-venous differences to study macronutrient metabolism: introduction and overview. AB - The arterio-venous difference technique is now well established in the study of organ and tissue metabolism. This technique requires samples to be obtained of the arterial blood supplying and the venous drainage from a tissue, together with a measurement of the blood flow through the tissue. The technique is most appropriate when the arterial concentration and tissue metabolism of a substance are constant, and when the blood flow is stable. If these criteria are not satisfied, care is needed in the interpretation of the results obtained. It should be recognized that the arterio-venous difference technique only measures the net exchange of a substance with the tissue, and that tracers are needed if unidirectional flux needs to be estimated. The other factors which must be borne in mind when intending to use this technique are the transit times of blood and the substance of interest through a tissue, the volume of distribution of the substance in the tissue, and the possibility that the venous samples obtained are derived from a mixture of different tissues. PMID- 10817155 TI - Macronutrient metabolism of adipose tissue at rest and during exercise: a methodological viewpoint. AB - The metabolism of white adipose tissue is regulated by many factors, including hormones and substrates delivered in the blood, the activity of the autonomic nervous system and the rate of flow of blood through the tissue. An integrated view of adipose tissue metabolism can only be gained, therefore, from studies in vivo. Of the various techniques available for studying adipose tissue metabolism in vivo, the measurement of arterio-venous differences offers some unique possibilities. In human subjects this technique has been performed mostly by catheterization of the venous drainage of the subcutaneous abdominal depot. Studies using this technique indicate that adipose tissue has an active pattern of metabolism, responding rapidly to meal ingestion by suppressing the release of non-esterified fatty acids, or to exercise with an increase in fat mobilization. Adipose tissue blood flow may also change rapidly in these situations; for instance, it increases markedly after a meal, potentially increasing the delivery of triacylglycerol to the enzyme lipoprotein lipase (EC 3.1.1.34) for hydrolysis. During exercise, there is evidence that adipose tissue blood flow does not increase sufficiently to allow delivery of all the fatty acids released into the systemic circulation. The various adipose tissue depots have their own characteristic metabolic properties, although in human subjects these are difficult to study with the arterio-venous difference technique. A combination of tracer infusion with selective catheterization allows measurements of leg, splanchnic and non-splanchnic upper-body fat mobilization and triacylglycerol clearance. Development of such techniques may open up new possibilities in the future for obtaining an integrated picture of adipose tissue function and its depot-specific variations. PMID- 10817156 TI - Limb and skeletal muscle blood flow measurements at rest and during exercise in human subjects. AB - The aim of the present review is to present techniques used for measuring blood flow in human subjects and advice as to when they may be applicable. Since blood flow is required to estimate substrate fluxes, energy turnover and metabolic rate of skeletal muscle, accurate measurements of blood flow are of extreme importance. Several techniques have therefore been developed to enable estimates to be made of the arterial inflow to, venous outflow from, or local blood flow within the muscle. Regional measurements have been performed using electromagnetic flow meters, plethysmography, indicator methods (e.g. thermodilution and indo-cyanine green dye infusion), ultrasound Doppler, and magnetic resonance velocity imaging. Local estimates have been made using 133Xe clearance, microdialysis, near i.r. spectroscopy, positron emission tomography and laser Doppler. In principle, the aim of the study, the type of interventions and the limitations of each technique determine which method may be most appropriate. Ultrasound Doppler and continuous indo-cyanine green dye infusion gives the most accurate limb blood flow measurements at rest. Moreover, the ultrasound Doppler is unique, as it does not demand a steady-state, and because its high temporal resolution allows detection of normal physiological variations as well as continuous measurements during transitional states such as at onset of and in recovery from exercise. During steady-state exercise thermodilution can be used in addition to indo-cyanine green dye infusion and ultrasound Doppler, where the latter is restricted to exercise modes with a fixed vessel position. Magnetic resonance velocity imaging may in addition be used to determine blood flow within deep single vessels. Positron emission tomography seems to be the most promising tool for local skeletal muscle blood-flow measurements in relation to metabolic activity, although the mode and intensity of exercise will be restricted by the apparatus design. PMID- 10817157 TI - Skeletal muscle substrate metabolism during exercise: methodological considerations. AB - The aim of the present article is to evaluate critically the various methods employed in studies designed to quantify precisely skeletal muscle substrate utilization during exercise. In general, the pattern of substrate utilization during exercise can be described well from O2 uptake measurements and the respiratory exchange ratio. However, if the aim is to quantify limb or muscle metabolism, invasive measurements have to be carried out, such as the determination of blood flow, arterio-venous (a-v) difference measurements for O2 and relevant substrates, and biopsies of the active muscle. As many substrates and metabolites may be both taken up and released by muscle at rest and during exercise, isotopes can be used to determine uptake and/or release and also fractional uptake can be accounted for. Furthermore, the use of isotopes opens up further possibilities for the estimation of oxidation rates of various substrates. There are several methodological concerns to be aware of when studying the metabolic response to exercise in human subjects. These concerns include: (1) the muscle mass involved in the exercise is largely unknown (bicycle or treadmill). Moreover, whether the muscle sample obtained from a limb muscle and the substrate and metabolite concentrations are representative can be a problem; (2) the placement of the venous catheter can be critical, and it should be secured so that the blood sample represents blood from the active muscle with a minimum of contamination from other muscles and tissues; (3) the use of net limb glycerol release to estimate lipolysis is probably not valid (triacylglycerol utilization by muscle), since glycerol can be metabolized in skeletal muscle; (4) the precision of blood-borne substrate concentrations during exercise measured by a-v difference is hampered since they become very small due to the high blood flow. Recommendations are given in order to obtain more quantitative and conclusive data in studies investigating the regulatory mechanisms for substrate choice by muscle. PMID- 10817158 TI - Microdialysis: use in human exercise studies. AB - Microdialysis has been used for 25 years to study brain function in vivo. Recently, it has been developed for investigations on peripheral tissues. A microdialysis catheter is an artificial blood vessel system which can be placed in the extracellular space of various tissues such as adipose tissue and skeletal muscle in order to examine these tissues in situ. Molecules are collected from the tissue by the device and their true interstitial concentration can be estimated. Metabolically-active molecules can be delivered to the interstitial space through the microdialysis probe and their action on the tissue can be investigated locally without producing generalized effects. It is also possible to study local tissue blood flow with microdialysis by adding a flow marker (usually ethanol) to the microdialysis solvent. The microdialysis technique is particularly useful for studies of small and water-soluble molecules. A number of important observations on the in vivo regulation of lipolysis, carbohydrate metabolism and blood flow in human skeletal muscle and adipose tissue have been made recently using microdialysis. PMID- 10817159 TI - Microdialysis of skeletal muscle at rest. AB - Techniques in human skeletal muscle research are by necessity predominantly 'descriptive'. Microdialysis has raised high expectations that it could meet the demand for a method that allows 'mechanistic' investigations to be performed in human skeletal muscle. In the present review, some views are given on how well the initial expectations on the use of the microdialysis technique in skeletal muscle have been fulfilled, and the areas in which additional work is needed in order to validate microdialysis as an important metabolic technique in this tissue. The microdialysis catheter has been equated to an artificial blood vessel, which is introduced into the tissue. By means of this 'vessel' the concentrations of compounds in the interstitial space can be monitored. The concentration of substances in the collected samples is dependent on the rate of perfusate flow. When perfusate flow is slow enough to allow complete equilibration between interstitial and perfusate fluids, the concentration in the perfusate is maximal and identical to the interstitial concentration. Microdialysis data may be influenced by changes in blood flow, especially in instances where the tissue diffusivity limits the recovery in vivo, i.e. when recovery in vitro is 100%, whereas the recovery in vivo is less than 100%. Microdialysis data indicate that a significant arterial-interstitial glucose concentration gradient exists in skeletal muscle but not in adipose tissue at rest. While the concentrations of glucose and lactate in the dialysate from skeletal muscle are close to the expected values, the glycerol values obtained for muscle are still puzzling. Ethanol added to the perfusate will be cleared by the tissue at a rate that is determined by the nutritive blood flow (the microdialysis ethanol technique). It is concluded that microdialysis of skeletal muscle has become an important technique for mechanistic studies in human metabolism and nutrition. PMID- 10817160 TI - Vasoactive substances in the interstitium of contracting skeletal muscle examined by microdialysis. AB - In the study of the regulation of skeletal muscle blood flow during exercise it is useful to obtain information regarding the concentrations of vasoactive substances in the muscle interstitium, a site where the compounds act on the vascular and skeletal muscle cells. The microdialysis technique is a useful tool for measuring interstitial substances in the muscle at rest and during exercise in human subjects, and the technique can also be used to study the effect of both systemic and local interventions in a specific area of an exercising muscle. Probe recovery, which represents the relative amount of a substance that is diffusing to the dialysis membrane, changes from rest to exercise and can be determined by the internal-standard technique which allows for a relatively high time resolution (min). Furthermore, the use of electrodes at the microdialysis outlet makes it possible to perform continuous measurements of interstitial substances. The present review gives examples of how the microdialysis technique has been applied to study potentially important vasodilators such as adenosine, NO and K+ in human skeletal muscles and highlights areas for future research to establish the functional importance of these compounds. PMID- 10817161 TI - An introduction to the use of tracers in nutrition and metabolism. AB - The present article is a review written at a level suitable for students and new workers to the field of techniques in common current use for the measurement of static and dynamic features of metabolism, especially nutritional metabolism. It covers the nature of radioactive and stable-isotope tracers, the means of measuring them, and the advantages and disadvantages of their use. The greater part of the review deals with methods for the measurement of pool sizes and metabolic processes, with the emphasis being on protein metabolism, a field the author knows best. The examples given are from a variety of sources, including the work of the author, but the principles underlying the techniques are universally applicable to all metabolic investigations using tracers. PMID- 10817162 TI - Body composition, water turnover and energy turnover assessment with labelled water. AB - Our understanding of human energy metabolism has benefited greatly from the application of water labelled with 2H and 18O for the measurement of total body water, water turnover and total daily energy expenditure. Applications include validation of techniques for the assessment of dietary intake and physical activity, assessment of water and energy requirement and the assessment of the effect of dietary and physical activity interventions, including its use with endurance athletes competing at the highest level. Critical aspects of the application are isotope dose preparation, sample collection, sample analysis and the calculation procedure. The labelled-water method can easily be applied in normal living conditions, including exercise, and in the clinical setting. However, sample analysis requires a sophisticated laboratory with an isotope ratio mass spectrometer and a sample preparation system. Examples of insights based on labelled-water studies are: (1) self-reported dietary intakes often underestimate energy requirements; (2) subjects have problems maintaining energy balance when daily energy expenditure exceeds 2.5 times resting energy expenditure. Devices for the assessment of physical activity validated using labelled water allow the study of activity patterns and strategies to influence the activity level of a sedentary society. PMID- 10817163 TI - Use of stable isotopes to study carbohydrate and fat metabolism at the whole-body level. AB - The present review discusses the advantages and limitations of using stable isotope tracers to assess carbohydrate and fat metabolism at the whole-body level. One advantage of stable- (v. radioactive-) isotope tracers is the relative ease with which the location of a label within a molecule can be determined using selected-ion-monitoring GC-mass spectrometry (SIM-GC-MS). This technique minimizes potential problems due to label recycling, allows the use of multiple labelled compounds simultaneously (e.g. to quantify glucose cycling), and perhaps most importantly, has led to the development of unique stable-isotope methods for, for example, quantifying gluconeogenesis. However, the limited sensitivity of SIM-GC-MS sometimes requires that relatively large amounts of a stable-isotope tracer be used, thus increasing cost and potentially altering metabolism. At least theoretically, stable- (or radioactive-) isotope tracers can also be used in conjunction with indirect calorimetry to estimate utilization of muscle glycogen or triacylglycerol stores, thus potentially circumventing the need to obtain muscle biopsies. These calculations, however, require certain critical assumptions, which if incorrect could lead to major errors in the values obtained. Despite such limitations, stable-isotope tracers provide a powerful and sometimes unique tool for investigating carbohydrate and fat metabolism at the whole-body level. With continuing advances in availability, instrumentation and methods, it is likely that stable-isotope tracers will become increasingly important in the immediate future. PMID- 10817164 TI - Quantifying the contribution of gluconeogenesis to glucose production in fasted human subjects using stable isotopes. AB - The contribution of gluconeogenesis to glucose production is estimated from the enrichment of the H bound to C-5 of glucose relative to either that bound to C-2 of glucose or the enrichment in body water on ingesting 2H2O in the fasted state. Contributions of all gluconeogenic substrates are included in the estimate and the limitation of an uncertain precursor enrichment removed. The half-life of 2H2O in body water precludes a repeat study for many weeks. Glycogen cycling could result in underestimation, but there is evidence that glycogen cycling does not occur in liver in the fasted state. Gluconeogenesis has been estimated by mass-isotopomer-distribution analyses, usually by administering 13C-labelled glycerol. Underestimates emphasize the major limitation of the method, i.e. the need to assume a single enrichment of the precursor pool. Estimates of gluconeogenesis from isotopomer distribution in arterial-blood glucose and lactate on infusing [U-13C6] glucose are unreliable, as a proportion of the glucose is formed from glycerol and from amino acids not converted to glucose via pyruvate. Loss of label in the Krebs cycle and relying on enrichment of arterial blood lactate as a measure of hepatic pyruvate further add to the uncertainty. Estimates of the rate of gluconeogenesis by NMR are obtained by subtraction of the rate of glycogenolysis determined by NMR from the rate of glucose production. Estimates are then the mean rate for the period over which glycogen contents are measured. Technical considerations can limit the accuracy of analyses and result in overestimates. PMID- 10817165 TI - Glycerol production and utilization measured using stable isotopes. AB - The rate of appearance of glycerol in the systemic circulation is determined from the enrichment of arterial blood glycerol when labelled glycerol is infused intravenously. This value provides a good measure of whole-body lipolysis during fasting, except that arterial infusion and venous sampling, if feasible, would probably give a higher more-accurate value. Lipolysis occurs primarily in adipose tissue, although other tissues contribute, notably muscle. Measurement is based on the difference in the enrichment of the glycerol entering and leaving the tissue. Lipolysis is underestimated by the extent to which glycerol released by lipolysis does not enter the systemic circulation, as occurs when lipolysis takes place in the non-hepatic tissue of the splanchnic bed. Glycerol released into the systemic circulation is utilized mainly by liver, although kidney and muscle are also major users of glycerol. Measurement of glycerol utilization is based on the amount of labelled glycerol taken up by the tissues. Other tissues probably utilize glycerol to a smaller extent, but in total this represents a significant amount. Most glycerol taken up by liver is converted to glucose. Glucose is probably the major source of glycerol-3-phosphate used in the esterification of fatty acids by adipose tissue. PMID- 10817166 TI - Correction factors for 13C-labelled substrate oxidation at whole-body and muscle level. AB - The oxidation of fatty acids, carbohydrates and amino acids can be measured by quantifying the rate of excretion of labelled CO2 following administration of 14C or 13C-labelled substrates at whole-body and tissue level. However, there is a theoretical need to correct the oxidation rates for the proportion of labelled CO2 that is produced via oxidation but not excreted. Furthermore, depending on the substrate and position of the C label(s), there may also be a need to correct for labelled C from the metabolized substrate that does not appear as CO2, but rather becomes temporarily fixed in other metabolites. The bicarbonate correction factor is used to correct for the labelled CO2 not excreted. Recently, an acetate correction factor has been proposed for the simultaneous correction of CO2 not excreted and label fixed in other metabolites via isotopic exchange reactions, mainly in the tricarboxylic acid cycle. Changes in metabolic rate induced, for example, by feeding, hormonal changes and physical activity, as well as infusion time, have been shown to affect both correction factors. The present paper explains the theoretical and physiological basis of these correction factors and makes recommendations as to how these correction factors should be used in various physiological conditions. PMID- 10817167 TI - Tracers to investigate protein and amino acid metabolism in human subjects. AB - Three tracer methods have been used to measure protein synthesis, protein breakdown and protein oxidation at whole-body level. The method using L-[1 (13)C]leucine is considered the method of reference. These methods have contributed greatly to the existing knowledge on whole-body protein turnover and its regulation by feeding, fasting, hormones and disease. How exercise and ingestion of mixed protein-containing meals affect whole-body protein metabolism is still open to debate, as there are discrepancies in results obtained with different tracers. The contribution of whole-body methods to the future gain of knowledge is expected to be limited due to the fact that most physiological disturbances have been investigated extensively, and due to the lack of information on the relative contribution of various tissues and proteins to whole body changes. Tracer amino acid-incorporation methods are most suited to investigate these latter aspects of protein metabolism. These methods have shown that some tissues (liver and gut) have much higher turnover rates and deposit much more protein than others (muscle). Massive differences also exist between the fractional synthesis rates of individual proteins. The incorporation methods have been properly validated, although minor disagreements remain on the identity of the true precursor pool (the enrichment of which should be used in the calculations). Arterio-venous organ balance studies have shown that little protein is deposited in skeletal muscle following a protein-containing meal, while much more protein is deposited in liver and gut. The amount deposited in the feeding period in each of these tissues is released again during overnight fasting. The addition of tracers to organ balance studies allows the simultaneous estimation of protein synthesis and protein breakdown, and provides information on whether changes in net protein balance are caused primarily by a change in protein synthesis or in protein breakdown. In the case of a small arterio-venous difference in a tissue with a high blood flow, estimates of protein synthesis and breakdown become very uncertain, limiting the value of using the tracer. An additional measurement of the intracellular free amino acid pool enrichment allows a correction for amino acid recycling and quantification of the inward and outward transmembrane transport. However, in order to obtain reliable estimates of the intramuscular amino acid enrichment and, therefore, of muscle protein synthesis and breakdown in this so-called three-pool model, the muscle should be freeze-dried and the resulting fibres should be freed from connective tissue and small blood clots under a dissection microscope. Even when optimal precautions are taken, the calculations in these tracer balance methods use multiple variables and, therefore, are bound to lead to more variability in estimates of protein synthesis than the tracer amino acid incorporation methods. In the future, most studies should focus on the measurement of protein synthesis and breakdown in specific proteins in order to understand the mechanisms behind tissue adaptation in response to various stimuli (feeding, fasting, exercise, trauma, sepsis, disuse and disease). The tracer laboratories, therefore, should improve the methodology to allow the measurement of low tracer amino acid enrichments in small amounts of protein. PMID- 10817168 TI - An overview of methods for assessment of free radical activity in biology. AB - Assays which purport to assess free radical activity in biological systems are multiple. However, despite numerous published descriptions of new methods and modifications of methods to assess free radical activity in biological materials, there is still a lack of reliable techniques for quantification of activity in vivo. Analysis of a number of related indicators and use of a variety of approaches appears the only reliable way to evaluate these processes in vivo. In studies of free radical generation by contracting skeletal muscle we have attempted to use a variety of indicators, including measurement of endogenous antioxidant levels, measurement of indirect indicators of free radical activity (e.g. products of lipid peroxidation, DNA oxidation or protein oxidation) and, where possible, measurement of direct indicators of free radical activity by electron spin resonance techniques. In view of the relative lack of specificity of many available techniques, caution should be exerted in evaluating the numerous examples of isolated single measures of free radical activity which are present in the scientific literature. PMID- 10817169 TI - Methods to detect DNA damage by free radicals: relation to exercise. AB - Epidemiological investigations repeatedly show decreased morbidity from regular exercise compared with sedentary life. A large number of investigations have demonstrated increased oxidation of important cellular macromolecules, whereas other investigators have found no effects or even signs of lowering of oxidation of macromolecules. In particular, extreme and long-duration strenuous exercise appears to lead to deleterious oxidation of cellular macromolecules. The oxidation of DNA is important because the oxidative modifications of DNA bases, particularly the 8-hydroxylation of guanine, are mutagenic and have been implicated in a variety of diseases such as ageing and cancer. The methodologies for further investigation of the relationship between DNA oxidation and exercise are available. The preferred methods rely on HPLC or GC-mass spectrometry; whereas the theoretically-attractive liquid chromatography-tandem mass spectrometry is being developed. Caution should be taken to avoid artifacts because of the six orders of magnitude of difference between oxidized and non oxidized DNA bases in tissues. The methods can be used to estimate tissue levels, i.e. a local concentration of oxidized DNA, or to estimate the rate of body DNA oxidation by the urinary output of repair products, the latter being a method that is independent of repair. During exercise there appears to be a shifting of dietary-dependent antioxidant, e.g. vitamin C and vitamin E, from muscle to plasma, and an increased oxidation in plasma of these antioxidants. Supplementation trials with antioxidants have not been able to increase exercise performance; however, optimum nutrition with antioxidants and possibly supplementation, could be important in the prevention of diseases in the long term. The pattern from these observations appears to be quite consistent; immediately after exercise, regardless of how intense, there do not appear to be any signs of oxidative damage to DNA. Acute or prolonged moderate exercise does not produce signs of oxidative DNA damage and might even be associated with lowering of the levels of oxidation of tissue DNA; however, after long-duration and intense exercise an increase in oxidative DNA modifications is apparent. We suggest as a hypothesis that the relationship between exercise and health is U shaped. This hypothesis needs to be tested in detail in order to establish the maximum beneficial exercise level with regard to oxidative DNA modification, and also the level that could be deleterious and might even increase the risk for cancer and other diseases. PMID- 10817170 TI - Determination of activity of antioxidants in human subjects. AB - Evidence from biochemical and animal models suggests that nutritional antioxidants should inhibit the development of diseases such as CHD and certain cancers. This evidence is not clearly corroborated by intervention studies in human subjects, due, in part, to inadequacies in current analytical methodologies. Although in vitro assays can give useful information on the attributes required by a compound to act as an antioxidant, results may have little nutritional relevance due to limited bioavailability. The determination of antioxidants in blood is often used as a measure of antioxidant status in vivo, but may not necessarily reflect concentrations in target tissues where oxidative stress is greatest. In addition, the accumulation of antioxidants in selective tissues may not be apparent from plasma measurements. Participation in quality control schemes for antioxidant determination by HPLC allows inter-laboratory comparison of results. Moderation of indices of oxidative damage to lipids, proteins and DNA can provide information on the effectiveness of compounds as nutritional antioxidants. However, most current methods of assessing oxidative stress are subject to confounding factors of non-oxidative origin. Assays for total antioxidant capacity in plasma differ in their type of oxidation source, target and measurement used to detect the oxidized product. They give different results, should never be used in isolation, and results should be interpreted with caution. Until more is known about the activity and metabolic fate of antioxidants, caution should be exercised in the consumption of large amounts of commercially-available antioxidant preparations. PMID- 10817171 TI - Analysis of cellular responses to free radicals: focus on exercise and skeletal muscle. AB - Muscular exercise results in an increased production of radicals and other forms of reactive oxygen species (ROS). Recent evidence suggests that radicals and other ROS are an underlying aetiology in exercise-induced disturbances in muscle redox status. These exercise-induced redox disturbances in skeletal muscle are postulated to contribute to both muscle fatigue and/or exercise-induced muscle injury. To defend against ROS, muscle cells contain complex cellular defence mechanisms to reduce the risk of oxidative injury. Two major classes (enzymic and non-enzymic) of endogenous protective mechanisms work together to reduce the harmful effects of oxidants in the cell. Primary antioxidant enzymes include superoxide dismutase (EC 1.15.1.1; SOD), GSH peroxidase (EC 1.11.1.9; GPX), and catalase (EC 1.11.1.6); these enzymes are responsible for removing superoxide radicals, H2O2 and organic hydroperoxides, and H2O2 respectively. Important non enzymic antioxidants include vitamins E and C, beta-carotene, GSH and ubiquinones. Vitamin E, beta-carotene and ubiquinone are located in lipid regions of the cell, whereas GSH and vitamin C are in aqueous compartments of the cell. Regular endurance training promotes an increase in both total SOD and GPX activity in actively-recruited skeletal muscles. High-intensity exercise training has been shown to be generally superior to low-intensity exercise in the upregulation of muscle SOD and GPX activities. Also, training-induced upregulation of antioxidant enzymes is limited to highly-oxidative skeletal muscles. The effects of endurance training on non-enzymic antioxidants remain a relatively uninvestigated area. PMID- 10817172 TI - Quality of life and surgical outcome after laparoscopic Nissen and Toupet fundoplication: one-year follow-up. AB - BACKGROUND AND STUDY AIMS: Quality of life data are becoming widely accepted as a measure of surgical outcome, but the multifaceted symptoms in patients with gastrointestinal disorders are a challenge for this type of evaluation. The aim of the present study was to determine any potential differences in quality of life, specifically in patients undergoing either laparoscopic "floppy" Nissen fundoplication or Toupet fundoplication. PATIENTS AND METHODS: Using the Gastrointestinal Quality of Life Index (GIQLI), the quality of life data for 175 consecutive patients undergoing laparoscopic "floppy" Nissen (n=107) or Toupet (n=68) fundoplication at our department of surgery over a period of 30 months were evaluated prospectively. The patients included 97 men and 78 women, with a mean age of 52 years. The GIQLI creates a general score for quality of life by classifying five different subscales: gastrointestinal symptoms, emotional status, physical and social functions, and stress of medical treatment. This questionnaire was given to the patients preoperatively, and on three occasions after surgery -- at six weeks, three months, and one year. RESULTS: The analysis showed that the patients had a low GIQLI preoperatively in comparison with healthy individuals (mean 90.4 vs. 122.6 points), with all subscales being affected. The general score improved significantly six weeks postoperatively (mean: 118.2 points; P<0.05), showed further improvement at three months (mean: 124.2 points), and remained stable at one year (mean: 123.1 points; P<0.01) postoperatively. There were no differences in the quality of life or side effects between patients with a Nissen or Toupet fundoplication, except regarding the frequency of mild, transient dysphagia. CONCLUSIONS: Patients with gastroesophageal reflux disease suffer from a poor quality of life. After laparoscopic fundoplication, the quality of life improves and becomes comparable to that of healthy individuals. In our view, quality of life data should be evaluated as a major factor in determining the role of surgical interventions. Patients should receive consultation and advice regarding quality of life questions prior to surgery. PMID- 10817173 TI - Diagnostic yield and effect on clinical outcomes of push enteroscopy in suspected small-bowel bleeding. AB - BACKGROUND AND STUDY AIMS: Endoscopic examination of the small bowel (enteroscopy) has become an important modality in the investigation of patients with obscure gastrointestinal bleeding/anaemia. The aim of this study was to look at our clinical experience and also to determine the clinical outcomes following push enteroscopy (PE) in patients with suspected small intestinal bleeding. PATIENTS AND METHODS: The indications for PE were iron-deficiency anaemia/occult or obscure gastrointestinal bleeding (50 patients) and overt gastrointestinal bleeding (28 patients). Out of these 78 patients, a subgroup of 21 unselected patients with suspected small intestinal bleeding (six men, 15 women; mean age 59, range 26-85) was recruited to determine the clinical outcomes following PE. The principal outcome measures in this study were a) whether the procedure led to a change in the management of the patients referred for PE and b) whether it saved them from having further investigations. RESULTS: In those patients with undiagnosed gastrointestinal bleeding/anaemia, small-bowel arteriovenous malformations (AVMs) were detected in 24/78 (31%). Heater probe ablation therapy was performed in 23 patients with small-bowel and gastric AVMs. In total, seven of these 23 patients (30%) re-bled or required transfusion; these seven had repeat enteroscopy and heater probe ablation therapy. Three of these seven re bled and required further enteroscopy and therapy. PE resulted in a change in management and treatment plans in eight of 20 patients (40%). The certainty of diagnosis increased in seven of 20 patients (35%). It increased from a mean value of 1.35 before PE to 2.40 following PE ( P = 0.01). The test was assigned a median "usefulness score" of 3 by the requesting physicians on an integer scale from 1 (not helpful) to 5 (very helpful). CONCLUSIONS: We conclude that (i) PE can establish a diagnosis in a high proportion of patients with undiagnosed gastrointestinal bleeding/anaemia; (ii) heater probe ablation therapy of vascular lesions can be performed routinely at the time of PE; (iii) a significant proportion of patients (42%) referred for enteroscopy have lesions in the stomach/proximal duodenum that are missed at diagnostic endoscopy; (iv) PE provides helpful information in routine clinical practice and is perceived as a helpful test by requesting physicians; (v) PE improves certainty of diagnosis as perceived by requesting physicians, and plays an important role in altering management plans. PMID- 10817174 TI - Does gastroscopy induce myocardial ischemia in patients with coronary heart disease? AB - BACKGROUND AND STUDY AIMS: Gastroscopy has been reported to be dangerous for unstable patients with coronary heart disease (CHD). The aims of this study were to find out whether endoscopy is equally liable to cause myocardial ischemia in stable CHD patients, and whether this can be predicted prior to endoscopy, and to find out the frequency of abnormal findings in patients for whom a secondary prophylaxis with acetylsalicylic acid (ASA) is indicated. PATIENTS AND METHODS: Electrocardiograph recording using a Holter monitor was performed during gastroscopy in 71 patients with stable CHD, to check for silent ischemia. To predict potential ischemia during gastroscopy, the Holter monitoring ECG was applied prior to a treadmill test, and withdrawn after gastroscopy 16-22 hours later. RESULTS: During gastroscopy, 30 patients (42%) had silent ischemia, but only 1 patient (1%) became symptomatic. Ischemia was dependent on heart-rate (median heart rate with ischemia 124 beats/min, without 104 beats/min). Abnormal findings on gastroscopy were found in 53 patients (75%). They implied a potential bleeding risk in 30 patients (42%) and prevented the indication for ASA in 6 of them (8%). CONCLUSIONS: Gastroscopy is potentially a harmful procedure for CHD patients, but the incidence of ischemic periods may be reduced by conscious sedation and, if the patient is receiving beta-blocking agent therapy, by applying this medication prior to gastroscopy. PMID- 10817175 TI - High complication rate of bile duct stents in patients with chronic alcoholic pancreatitis due to noncompliance. AB - BACKGROUND AND STUDY AIMS: Biliary obstruction in chronic pancreatitis is frequently treated by endoscopic insertion of a plastic stent into the common bile duct, a therapy regarded as having a low complication rate. The aim of this study is to analyze the frequency and severity of complications caused by biliary stents in patients with chronic alcoholic pancreatitis. PATIENTS AND METHODS: We retrospectively analyzed all our patients with chronic pancreatitis (n = 14) who were provided with a plastic stent for biliary stenosis between June 1993 and December 1997. Stent exchanges were followed until December 1998. RESULTS: Stent insertion was performed without early complications and was successful in each patient. Only two patients were admitted after 3-4 months at the scheduled dates for stent exchange, both without complications. In one of these patients, the bile duct stenosis was reopened after two stent exchanges over a total period of 8 months. Most of our patients (n=12) did not come at the arranged dates for stent exchange. They were repeatedly admitted (mean 2.9 times/patient, range 1-5) as emergency cases with severe complications of biliary obstruction, such as cholangitis or biliary sepsis. Reopening of the bile duct stenosis was not achieved in these patients. CONCLUSIONS: We associate the high rate of complications with the noncompliance of our patients, who were all alcoholics. The high incidence of late complications in noncompliant patients is a limitation of biliary stenting, and appears to be potentially harmful. PMID- 10817176 TI - Technical considerations and patient comfort in total colonoscopy with and without a transparent cap: initial experiences from a pilot study. AB - BACKGROUND AND STUDY AIMS: Even though colonoscopy was introduced 30 years ago, endoscopists still leave a small percentage of the colonic mucosal surface unexamined because of the limitations of the procedure, so there is still room for technical improvements. The aim of this pilot study was to test the feasibility of attaching a transparent cap to the tip of the colonoscope, partly to gather basic data and experience for planning a larger randomized study, and partly to evaluate any technical advantages or disadvantages, and also to appraise the patients' experiences. PATIENTS AND METHODS: In 50 consecutive patients examined by a single experienced endoscopist, a prospective pilot study was carried out comparing the use of a colonoscope fitted with a transparent cap (in half of the colonoscopies) with the use of an ordinary colonoscope without a cap (in the remaining 25 patients). The following parameters were recorded: indication for colonoscopy, time to reach the cecum, total time for the colonoscopy, findings, diagnosis, type of colonoscopy (diagnostic or therapeutic, partial or total, ileal intubation), the amount of analgesia and sedation given during the endoscopy, and complications. At the end of the examination and also before leaving the hospital, the patients were asked by a nurse to estimate pain experienced during the colonoscopy, using a visual analogue scale. RESULTS: The time for the procedure, the ability to perform a complete colonoscopy (including ileal intubation when it was medically relevant), and the complication rate turned out to be similar in both groups. There were no differences between the amounts of analgesia and sedation given during the endoscopy or between the patients' estimations of the pain experienced. CONCLUSIONS: The time to reach the cecum and the total time for the colonoscopy is the same with or without the cap, which is well tolerated by patients. Using the cap greatly facilitates the possibility of finding small polyps behind folds in the colon, because the folds can be straightened, thereby improving the view, although in this small pilot study it could not be proven that the number of polyps found was greater using a cap-fitted colonoscope. PMID- 10817177 TI - Dilation of benign strictures following low anterior resection using Savary Gilliard bougies. AB - BACKGROUND AND STUDY AIMS: Benign anastomotic strictures occur in up to 22% of patients after colorectal resections. Initially, treatment for these strictures was surgical, but nowadays endoscopic dilation techniques are preferred. This study was conducted to assess the efficacy of dilation using SavaryGilliard bougies. PATIENTS AND METHODS: From 1987 to 1994, 256 consecutive patients underwent low anterior resection (LAR). Twenty-one patients (8.2%) developed a stricture of the colorectal anastomosis. Follow-up data were available for 18 of these patients. The patients were treated using endoscopic Savary dilation, with bougies of increasing diameters (10-19 mm). The mean follow-up period was 19 months (1-60 months). RESULTS: Stricture symptoms presented after a mean period of 7.7 months after LAR. In three of the 18 patients, the stenosis was caused by local recurrence, and these patients were excluded from further evaluation. Normal defecation was restored in 10 of the remaining 15 patients, and symptoms disappeared. In five patients, there was only partial improvement, but only three of them required another type of treatment. Of four patients who received radiotherapy and developed a strictured anastomosis, two had successful dilations. A normal defecation pattern was never regained if more than three dilations were necessary. No complications caused by Savary dilation were observed. CONCLUSIONS: In this study population, Savary dilation appeared to be a safe and effective treatment for benign anastomotic strictures after LAR. All successfully treated patients (ten of 15) required no more than three dilations. Two other patients had partial success. Only three patients required another form of treatment (two endoscopic, one surgical). PMID- 10817179 TI - Endoscopic ultrasound features of protein-losing gastropathy with hypertrophic gastric folds. AB - BACKGROUND AND STUDY AIMS: We aim to clarify the endoscopic ultrasound (EUS) features of protein-losing gastropathy with hypertrophic gastric folds (PLGH), including Menetrier's disease. PATIENTS AND METHODS: We analyzed the EUS and histologic findings in five patients who underwent both endoscopic ultrasonography and endoscopic resection. RESULTS: Histologically, we diagnosed one patient as having acute gastritis, three patients as having Menetrier's disease, and the remaining patient as having hypertrophic lymphocytic gastritis (HLG). Helicobacter pylori was recognized in all but one patient. At EUS every patient was found to have giant gastric folds (13 to 20 mm in diameter), resulting from echogenic thickening of the mucosal layer with or without cystic components. Two patients who underwent eradication therapy of H. pylori showed both clinical and morphologic resolution. CONCLUSIONS: Echogenic thickening of the mucosal layer may be a characteristic EUS feature of protein-losing gastropathy with hypertrophic gastric folds, and H. pylori may be one of the causative agents. PMID- 10817178 TI - Mucin-hypersecreting bile duct tumor characterized by a striking homology with an intraductal papillary mucinous tumor (IPMT) of the pancreas. AB - BACKGROUND AND STUDY AIMS: The mucin-hypersecreting bile duct tumor, which closely resembles an intraductal papillary mucinous tumor (IPMT) of the pancreas, is rare, and its clinical features are not well known. We report our experience of nine patients with this type of tumor, and analyze the data in order to elucidate its clinicopathologic characteristics. PATIENTS AND METHODS: Between 1995 and 1998, nine consecutive patients (four men, five women; mean age 54) who were diagnosed as having a mucin-hypersecreting bile duct tumor were enrolled in this study. RESULTS: Recent or previous attacks of biliary pain and acute cholangitis were reported by most of the patients. Characteristics included a widely open ampulla of Vater with extrusion of mucin, and a diffuse dilated intrahepatic and extrahepatic bile duct with amorphous filling defects on cholangiogram. On cholangioscopic examination, a papillary mass or minute mucosal lesion was found in the dilated bile duct containing thick viscid mucin. Nine patients were referred for an operation, and curative resection was performed in eight of them. Using histologic examination, well differentiated adenocarcinoma in the background of benign hyperplasia and adenoma was documented in all patients except one, who showed pure adenoma. CONCLUSION: The mucin hypersecreting bile duct tumor can be characterized by a striking homology with IPMT of the pancreas in clinical, radiologic, and pathologic features. PMID- 10817180 TI - A 47-year-old woman with pancreatitis and early satiety. PMID- 10817181 TI - Randomization is not the (only) answer: a plea for structured objective evaluation of endoscopic therapy. AB - Endoscopic therapy clearly has a primary role in many clinical conditions (such as dysphagia, obstructive jaundice, bleeding, and colonic polyps). There is much less certainty about the role of endoscopy in many other clinical contexts (e.g. management of chronic pancreatitis). Randomization is the gold standard for evaluation of competing therapies. Unfortunately, there are many difficulties in mounting meaningful randomized controlled trials of endoscopic methods. Many have been done, but few have provided us with real practical answers. This article argues that the evidence we need to advise patients is often better obtained through very stringent observational studies, provided that all necessary data elements are defined and documented and independent objective arbiters (referees) are fully involved in the process. PMID- 10817182 TI - Magnetic resonance imaging in gastroenterology: time to say good-bye to all that endoscopy? AB - Magnetic resonance imaging (MRI) has been described as the most important development in medical diagnosis since the discovery of the roentgen ray more than 100 years ago. The effectiveness of MRI has been extended to make it applicable in a wide variety of gastrointestinal disorders. The attention of gastroenterologists is currently focusing on pancreaticobiliary and bowel diseases. Magnetic resonance cholangiopancreatography (MRCP) has become a competitive alternative to diagnostic endoscopic retrograde cholangiopancreatography in a variety of hepatobiliary and pancreatic diseases. Magnetic resonance enteroscopy has the potential to become the preferable method for evaluating the entire small bowel; virtual colonoscopy, on the other hand, is far from the stage at which it could be promoted as a tool for general screening purposes in suspected colonic diseases. Its drawbacks include problems with standardization, implementation of the techniques in generalized settings, and patient acceptance. PMID- 10817183 TI - The Second European Endoscopy Forum (Sintra, Portugal, 17-18 June 1999): twenty questions on the esophagogastric junction. PMID- 10817184 TI - Polypoid metastatic hepatocellular carcinoma of the esophagus occurring after endoscopic variceal band ligation. AB - This report describes a rare case of metastatic hepatocellular carcinoma (HCC) presenting as a polypoid mass in the lower esophagus after endoscopic variceal band ligation (EVL). A 56-year-old man underwent EVL for variceal bleeding in September 1993. He presented with dysphagia and tarry stool in December 1993. An endoscopic examination revealed a semipedunculated polypoid mass at the lower part of the esophagus, where EVL had been performed 3 months previously. The histologic examination at autopsy revealed that the polypoid mass consisted of metastatic HCC that had spread via the retrograde portal flow. PMID- 10817185 TI - Life-threatening intraabdominal arterial embolization after histoacryl injection for bleeding gastric ulcer. AB - N-butyl-cyanoacrylate (Histoacryl) injection has become the treatment of choice for acutely bleeding esophagogastric varices, and is the only effective option for endoscopic treatment of gastric varices. Recent reports confirm the ability of Histoacryl injection therapy to achieve immediate hemostasis in cases of gastric ulcer bleeding or Dieulafoy ulcer, where conventional endoscopic hemostatic treatment had failed. Although the overall safety record of Histoacryl injection has been relatively good, there have been scattered cases of serious complications. Here, we present two patients showing life-threatening intraabdominal arterial embolization after Histoacryl injection. They had chronic gastric ulcers with active arterial bleeding. In spite of attempts at hemostatic treatment, complete hemostasis was not achieved. We injected Histoacryl, diluted with Lipiodol, into bleeding gastric ulcers, resulting in successful hemostasis. Soon after the procedure, intraabdominal arterial embolization developed in both patients. One patient survived and the other died. Based on these experiences, we would like to warn gastrointestinal endoscopists to be alert to these fatal complications, and we propose that less diluted Histoacryl seems to be preferable in cases of bleeding peptic ulcers. PMID- 10817186 TI - Bedside endosonography and endosonography-guided fine-needle aspiration in critically ill patients: a way out of the deadlock? AB - Endosonography and endosonography-guided fine-needle aspiration (EUS-FNA) are now established diagnostic techniques, which are performed electively in endoscopy suites. We report here the bedside use of EUS-FNA in three critically ill patients in an intensive-care unit, with a significant impact on the outcome. A mediastinal abscess after percutaneous dilational tracheotomy was aspirated in one patient, leading to appropriate antibiotic therapy and complete recovery. A paratracheal hematoma compressing the right main bronchus was aspirated in a patient with polytrauma, relieving the pressure effects. The third patient, who had end-stage dilated cardiomyopathy and was being evaluated for cardiac transplantation, was found to have an apical lung lesion suspicious for bronchogenic carcinoma. EUS was performed to exclude mediastinal metastasis and allow simultaneous resection at the time of transplantation. Although a metastasis was excluded by EUS-FNA, the patient died while awaiting surgery. We conclude that bedside EUS-FNA is a feasible procedure, and in experienced hands it can offer an alternative in life-threatening situations. PMID- 10817187 TI - Hemosuccus pancreaticus--a rare cause of gastrointestinal bleeding: diagnosis and interventional radiological therapy. AB - Hemorrhage from the pancreatic duct, i.e. hemosuccus pancreaticus (HP), is a rare cause of gastrointestinal bleeding. Pancreatic hemosuccus is usually due to the rupture of an aneurysm of a visceral artery, most likely the splenic artery, in chronic pancreatitis. Other causes of HP are rare. We present a case of HP in a female patient with no history but with positive findings of chronic calcifying pancreatitis upon ultrasonographic investigation, computed tomography scan, and endoscopic retrograde cholangiopancreatography. With detectable fresh blood in the descending duodenum, angiography of the celiac artery revealed an aneurysm of the splenic artery as the suspected cause of intermittent bleeding from the pancreatic duct. The treatment is traditionally surgical or by interventional radiological means. This is the first case described in the literature in which interventional radiological therapy involved implantation of an uncoated metal Palmaz stent in the splenic artery. In the follow-up of 18 months no relapse of HP was observed. PMID- 10817188 TI - Is bleeding after endoscopic sphincterotomy always a problem? PMID- 10817189 TI - Guidelines of the French Society of Digestive Endoscopy: total colonoscopy indications. The Council of the French Society of Digestive Endoscopy (SFED). PMID- 10817190 TI - Duodenal ulcer perforation following cyanoacrylate injection. PMID- 10817191 TI - A new stent device (Choo stent) for palliation of malignant gastric outlet obstruction. PMID- 10817192 TI - An economically constructed feeding tube for pull-type percutaneous endoscopic gastrostomy. PMID- 10817193 TI - Simultaneous endoscopic variceal sclerotherapy and ligation (SSL) with a newly designed multiple band ligator system accepting a sclerotherapy needle through the working channel. PMID- 10817194 TI - Bronchoesophageal fistula as a complication of percutaneous endoscopic gastrostomy. PMID- 10817195 TI - Granulomatous hepatitis in an immunocompromised patient diagnosed by minilaparoscopy. PMID- 10817196 TI - Rectal fistula causing persistent watery diarrhea. PMID- 10817197 TI - Image of an esophageal intramural hematoma. PMID- 10817198 TI - Endoscopic diagnosis in typhoid fever. PMID- 10817199 TI - The nature of oxygen in sporopollenin from the pollen of Typha angustifolia L. AB - Native and peracetylated sporopollenin from the pollen of Typha angustifolia L. was investigated using several spectroscopic methods, inducing Fourier transform infrared spectroscopy (FTIR), solid-state 13C-nuclear magnetic resonance spectroscopy (13C-NMR) and X-ray photoelectron spectrometry (XPS). Interpretation of the experimental data shows that the greater part of oxygen found in sporopollenin originates from hydroxyl groups and must be derived from aliphatics and not from aromatics. This result indicates that not only aromatics and long unbranched aliphatics but also poly-hydroxyl aliphatic components are involved in the complex structure of the polymer. Furthermore, it is most probable that the monomers of the sporopollenin skeleton are linked by ether- and not by ester linkage. Two possible approaches are suggested for the characterisation of sporopollenin structure. PMID- 10817200 TI - Iridoid glucosides from Phlomis tuberosa L. and Phlomis herba-ventis L. AB - A new iridoid glucoside, 5-desoxysesamoside, was isolated from Phlomis tuberosa L. (Lamiaceae) together with three known iridoid glucosides sesamoside, shanziside methyl ester and lamalbid. Lamiide was found in P. herba-ventis ssp. pungens in high concentrations. PMID- 10817201 TI - A new cucurbitacin glycoside from Kageneckia oblonga (Rosaceae). AB - A novel cucurbitacin glycoside has been isolated from aerial parts of Kageneckia oblonga R. et P. and shown to be 3beta-(beta-D-glucosyloxy)-16alpha,23alpha epoxycuc urbita-5,24-dien-11-one. The structure was established by usual spectroscopic and two-dimensional (2D) NMR techniques. This compound has found to be nontoxic when tested in-vivo cell culture assays. In previous investigations we reported 23,24-dihydrocucurbitacin F and prunasine. This was the first report on cucurbitacins from the genus Kageneckia (Rosaceae). PMID- 10817202 TI - Succinopyoverdins--a new variety of the pyoverdin chromophore. AB - Pseudomonas spp. of the fluorescent group produce siderophores (so-called pyoverdins) consisting of a peptide chain attached to a pyrimidoquinoline ring system which is derived from a condensation product of L-Dab and D-Tyr. Commonly several related compounds are found to accompany the pyoverdins having the same peptide chain, but differing in the heterocyclic part. The structure elucidation of a new variety (succinopyoverdin) is described here. PMID- 10817203 TI - Can the peptide chain of a pyoverdin be bound by an ester bond to the chromophore?--The old problem of pseudobactin 7SR1. AB - The structure which had been proposed for the pyoverdin named pseudobactin 7SR1 (Yang and Leong, 1984) differed from those of all other pyoverdins investigated so far: its peptide chain was supposedly linked to the chromophore not by an amide bond originating from its N-terminal amino acid, but rather by an ester bond involving one of the three Ser. It will be shown that the peptide chain of pseudobactin 7SR1 is actually bound to the chromophore amidically by its N terminal Ser and that it comprises a cyclodepsipeptidic substructure with an ester bond between the C-terminal Thr and the OH-group of the second Ser in the chain. PMID- 10817204 TI - A new class of biflavonoids: 2'-hydroxygenistein dimers from the roots of white lupin. AB - Two novel isoflavonoid dimers presumably originating from 2'-hydroxygenistein, 5,7,4'-trihydroxycoumaranochroman-4-one-(3-->5"')-5",7",2"'4"'- tetrahydroxyisoflavone (1, lupinalbisone A) and 5,7,4'-trihydroxycoumaranochroman 4-one-(3-6")-5",7",2"',4"'-te trahydroxyisoflavone (2, lupinalbisone B) were isolated from the roots of Lupinus albus L., and their structures involving relative stereochemistry were elucidated by spectroscopic methods. Using horse radish peroxidase and 2'-hydroxygenistein (3) as the substrate revealed the formation of these dimers together with 5,7,4'-trihydroxycoumaronochromone (4, lupinalbin A). Dimerization of 3 caused a remarkable increase of antifungal activity. PMID- 10817205 TI - Systematic study of high molecular weight compounds in Amazonian plants by high temperature gas chromatography-mass spectrometry. AB - The fractions of hexane and dichloromethane extraction from marupa (Simaruba amara) and (Bertholletia excelsa) leaves were analyzed by HT-HRGC (high temperature high resolution gas chromatography) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds can be characterized including unusual high molecular weight compounds. PMID- 10817206 TI - Comparison of headspace techniques for sampling volatile natural products in a dynamic system. AB - Commonly used dynamic sorption techniques for collecting biologically active volatile compounds have been compared. Solid phase microextraction (SPME) using two types of fibers (polydimethylsiloxane, PDMS, 100 microm, and carbowax/divinylbenzene, CW/DVB, 65 microm) was compared to purge and trap methods (Porapak Q, Tenax TA and charcoal) and a technique based on absorption in methanol in a cooling bath. Sampling was done in a stream of purified air (20 ml/min) in a closed and temperature-regulated (27 degrees C) glass tube, passing over a capillary tube containing a hexane solution of tridecane, heptadecane, 1 octen-3-ol, 1-hexadecanol, ethyl tetradecanoate, alpha-pinene, linalool, terpinen 4-ol, cis-verbenol, verbenone, beta-caryophyllene, E,E-farnesol, and geranylgeraniol. With all of the methods, the sampling was performed for a period of 30 min before extraction and analysis was done on a GC-FID system. In general, SPME gave a higher response for all compounds except for alpha-pinene, which was only extracted by the CW/DVB fiber. Purge and trap methods and methanol absorption gave the same response for all substances extracted. None of the methods extracted hexadecanol and geranylgeraniol under the conditions used. However, the SPME equipped with the PDMS coating extracted heptadecane, E,E farnesol and ethyl tetradecanoate. Our results show that SPME, when selecting the fibers to fit the polarity and volatility of the compounds, is an outstanding extraction method compared to purge and trap and methanol absorption, especially for a qualitative analysis. The best conditions for storing fibers exposed to compounds of high volatility were at low temperatures (6 degrees C) in sealed vials, while the worst way was to leave the exposed fiber unprotected at room temperature (22 degrees C). The dynamic sampling system was effectively tested on a fruiting body of a polypore fungus (Ganoderma applanatum) emitting 1-octen-3 ol, and again SPME showed to be the most sensitive technique. PMID- 10817207 TI - Seeds of Trichosanthes kirilowii, an energy-rich diet. AB - The kernels of Trichosanthes kirilowii seeds contain a green oil which makes up for 62% of their dry matter. This oil consists up to 95% of triglycerides, 2% of glycolipids, 1.3% of phospholipids and 1.8% of chlorophylls. As fatty acid components the triglycerides, glycolipids and phospholipids contain the unsaturated fatty acids linoleic and oleic acid and the saturated palmitic acid. In the triglycerides 19% of the C18:3 acid occur with the configuration delta9 cis, delta11 trans, delta13 cis. This acid is called trichosanic acid and is absent in glycolipids and phospholipids which contain instead another C18:3 fatty acid, which has conjugated double bounds and occurs with an amount of 21% and 3%, respectively. Typically, these oil seeds contain in addition up to 30% of their dry matter proteins and up to 2.5% mono- and oligosaccharides. The monosaccharides consist of rhamnose, galactose and glucose and the oligosaccharides represent a mixture of tri- and tetrasaccharides. PMID- 10817208 TI - Two-dimensional TLC separation and mass spectrometric identification of anthraquinones isolated from the fungus Dermocybe sanguinea. AB - A new two-dimensional TLC technique was developed to separate substituted anthraquinones on silica plates using n-pentanol-pyridine-methanol (6:4:3, v/v/v) and toluene-ethyl acetate-ethanol-formic acid (10:8:1:2, v/v/v/v) as eluents. The good separation power of the new technique was demonstrated by applying it to the analysis of complex anthraquinone mixtures isolated from the Scandinavian Dermocybe sanguinea. Emodin, physcion, endocrocin, dermolutein, dermorubin, 5 chlorodermorubin, emodin-1-beta-D-glucopyranoside, dermocybin-1-beta-D glucopyranoside and dermocybin, and five new, earlier in D. sanguinea unidentified compounds, 7-chloroemodin, 5,7-dichloroemodin, 5,7 dichloroendocrocin, 4-hydroxyaustrocorticone and austrocorticone, were separated and identified on the basis of Rf-values, UV/Vis spectra and mass spectra. PMID- 10817209 TI - Investigations into enzymes of nitrogen metabolism of the ectomycorrhizal basidiomycete, Suillus bovinus. AB - Axenic mycelia of the ectomycorrhizal basidiomycete, Suillus bovinus, were grown in liquid media under continuous aeration with compressed air at 25 degrees C in darkness. Provided with glucose as the only carbohydrate source, they produced similar amounts of dry weight with ammonia, with nitrate or with alanine, 60-80% more with glutamate or glutamine, but about 35% less with urea as the respectively only exogenous nitrogen source. In crude extracts of cells from NH4(+)-cultures, NADH-dependent glutamate dehydrogenase exhibited high aminating (688 nmol x mg protein(-1) x min(-1)) and low deaminating (21 nmol x mg protein( 1) x min(-1)) activities. Its Km-values for 2-oxoglutarate and for glutamate were 1.43 mM and 23.99 mM, respectively. pH-optimum for amination was about 7.2, that for deamination about 9.3. Glutamine synthetase activity was comparatively low (59 nmol x mg protein(-1) x min(-1)). Its affinity for glutamate was poor (Km = 23.7 mM), while that for the NH4+ replacing NH2OH was high (Km = 0.19 mM). pH optimum was found at 7.0. Glutamate synthase (= GOGAT) revealed similar low activity (62 nmol x mg protein(-1) x min(-1)), Km-values for glutamine and for 2 oxoglutarate of 2.82 mM and 0.28 mM, respectively, and pH-optimum around 8.0. Aspartate transaminase (= GOT) exhibited similar affinities for aspartate (Km = 2.55 mM) and for glutamate (Km = 3.13 mM), but clearly different Km-values for 2 oxoglutarate (1.46 mM) and for oxaloacetate (0.13 mM). Activity at optimum pH of about 8.0 was 506 nmol x mg protein(-1) x min(-1) for aspartate conversion, but only 39 nmol x mg protein(-1) x min(-1) at optimum pH of about 7.0 for glutamate conversion. Activity (599 nmol x mg protein(-1) x min(-1)), substrate affinities (Km for alanine = 6.30 mM, for 2-oxoglutarate = 0.45 mM) and pH-optimum (6.5-7.5) proved alanine transaminase (= GPT) also important in distribution of intracellular nitrogen. There was comparatively low activity of the obviously constitutive enzyme, urease, (42 nmol x mg protein(-1) x min(-1)) whose substrate affinity was rather high (Km = 0.56 mM). Nitrate reductase proved substrate induced; activity could only be measured after exposure of the mycelia to exogenous nitrate. Routes of entry of exogenous nitrogen and tentative significance of the various enzymes in cell metabolism are discussed. PMID- 10817210 TI - Use of immobilized Candida cells on xylitol production from sugarcane bagasse. AB - In this study we used the yeast Candida guilliermondii FTI 20037 immobilized by entrapment in Ca-alginate beads (2.5-3 mm diameter) for xylitol production from concentrated sugarcane bagasse hemicellulosic hydrolysate in a repeated batch system. The fermentation runs were carried out in 125- and 250-ml Erlenmeyer flasks placed in an orbital shaker at 30 degrees C and 200 rpm during 72 h, keeping constant the proportion between work volume and flask total volume. According to the results, cell viability was substantially high (98%) in all fermentative cycles. The values of parameters xylitol yield and volumetric productivity increased significantly with the reutilization of the immobilized biocatalysts. The highest values of xylitol final concentration (11.05 g/l), yield factor (0.47 g/g) and volumetric productivity (0.22 g/lh) were obtained in 250-ml Erlenmeyer flasks containing 80 ml of medium plus 20 ml of immobilized biocatalysts. The support used in this study (Ca-alginate) presented stability in the experimental conditions used. The results show that the use of immobilized cells is a promising approach for increasing the xylitol production rates. PMID- 10817211 TI - Novel spiciferone derivatives from the fungus Drechslera hawaiiensis isolated from the marine sponge Callyspongia aerizusa. AB - From the marine sponge Callyspongia aerizusa collected from the Sea of Bali, Indonesia, fungal isolates of Drechslera hawaiiensis were obtained. Culture filtrates of the fungus yielded four spiciferone derivatives which include spiciferone A (1) and B (2), and two other novel derivatives including spiciferol A (3) which is an alcohol congener of spiciferone A (1) and compound 4 which is an monocyclic spiciferone congener featuring a butoxyl side chain. The structures of the novel compounds were established on the basis of NMR spectroscopic (1H, 13C, COSY) and mass spectrometric (EIMS) data. PMID- 10817212 TI - Gene expression of Medicago sativa inoculated with Sinorhizobium meliloti as modulated by the xenobiotics cadmium and fluoranthene. AB - Alfalfa plants (Medicago sativa cv. Europe) inoculated with Sinorhizobium meliloti 2011 (formerly Rhizobium meliloti, de Lajudie et al., 1994) were cultivated for 14 days under standardized growth conditions in mineral medium with addition of the heavy metal cadmium or the polycyclic aromatic hydrocarbon fluoranthene. These xenobiotics significantly reduced the numbers of root nodules before any visible damage to the plant could be detected. EC10, EC50, and EC90 (effective concentrations reducing nodulation, shoot and root fresh weight by 10, 50, or 90% compared to the control without pollutant) were calculated. EC50 for cadmium ranged from 5.8 microM (nodulation) to more than 20 microM (root fresh weight). Testing fluoranthene resulted in an EC50 of 2.5 microg cm(-2) for nodulation, and EC50 values of more than 35 microg cm(-2) for shoot and root biomass production, indicating that the effect parameter nodulation is 10-fold more sensitive than shoot and root fresh weight. With mRNA differential display techniques the effects of both xenobiotics on gene expression in alfalfa root systems were studied. 37 differentially displayed transcripts were detected. Two of them, called DDMs1 and DDMs2, were confirmed by northern hybridization to be down-regulated in the presence of the xenobiotics. The expression of transcript DDMs1 was enhanced in alfalfa control plants inoculated with rhizobia, the transcript level was increased 2.5-3-fold compared to non-inoculated plants. This positive effect of nodulation was suppressed, partly by 35 microg cm(-2) fluoranthene and totally by 20 microM cadmium. The decrease in DDMs1 transcription was highly affected by the cadmium concentration with an EC50 of 5.9 microM. Compared to nodulation, almost identical EC10, EC50, and EC90 values were found for DDMs1 expression. Sequence analysis of DDMs1 revealed a significant overall homology (50% identity) to a hypothetical protein from Arabidopsis thaliana with high similarity to a copper transporting ATPase. High levels of transcript DDMs2 were observed in control plants with a 50% decrease in the xenobiotic-treated plants. DDMs2 gave a strong homology (82% identity) to the cytoplasmatic 60S ribosomal protein L18 from Arabidopsis thaliana. PMID- 10817213 TI - Metabolic transformation of the brassinosteroid 24-epi-castasterone by the cockroach Periplaneta americana. AB - After feeding of 24-epi-castasterone to the cockroach Periplaneta americana an organ-specific epimerization of the brassinosteroid to 2,24-diepi-castasterone could be detected in female insects. The metabolite being observed only in the ovaries and not in the testes of the insect was identified by GC/MS in comparison with a synthesized authentic sample. Contrary, 24-epi-brassinolide is not metabolized in the sexual organs of Periplaneta americana. This is the first evidence of a metabolic transformation of a brassinosteroid in insects. PMID- 10817214 TI - Effect of counterions on the influence of dodecyltrimethylammonium halides on thermotropic phase behaviour of phosphatidylcholine bilayers. AB - Effects of dodecyltrimethylammonium chloride (DTAC), dodecyltrimethylammonium bromide (DTAB) and dodecyltrimethylammonium iodide (DTAI) on thermotropic phase behaviour of phosphatidylcholine bilayers as well as on 1H NMR spectra were studied. In order to enhance the effect of counterions on water structure two series of experiments were performed. In the first one the surfactants were added to the water phase and in the other one directly to lipid phase (a mixed film was formed). The effects of particular surfactants on the main phase-transition temperature were more pronounced when they were added to the water phase (1st method) instead of the lipid phase (2nd method). Furthermore, in the case of the first method the transitions were found asymmetrical while in the second method nearly symmetrical. It is suggested that surfactant-poor and surfactant-rich domains are formed when surfactants are added to the water phase. PMID- 10817215 TI - Phospholipids in Mediterranean cephalopods. AB - Polar lipids of the cephalopods Eledone moschata, Sepia officinalis and Todarodes sagittatus mantle, represent 50.5%, 66.1% and 74.2% of wet tissue respectively. On the other hand the polar lipids of these three species of cephalopods constitute of 80.8%, 94.8% and 93.7% of phospholipids, respectively. The main phospholipids identified were phosphatidylcholine (52.2, 51.3 and 58.4% of total phospholipids respectively in the above mentioned species), phosphatidylethanolamine (18.1, 19.7 and 23.9%), sphingomyelin (10.7, 15.2 and 6.7%), lyso-phosphatidylcholine (3.1, 3.8 and 1.8%) and the unusual lipid ceramide aminoethylphosphonic acid (15.9, 10 and 9.2%). The 56.8% of phosphatidylcholine in Eledone moschata, the 46% in Sepia officinalis and the 74.1% in Todarodes sagittatus refer to the structure of 1,2-diacyl-glycerocholine and the remaining percentage refer to the structure of 1-o-alkyl-2-acyl glycerocholine or 1-o-alkyl-1-enyl-2-acyl-glycerocholine. The 87.2% of phosphatidylethanolamine in Eledone moschata, the 81% in Sepia officinalis and the 90.7% in Todarodes sagittatus refer to the structure of 1,2-diacyl glyceroethanolamine and the remaining percentage refer to the structure of 1-o alkyl-2-acyl-glyceroethanolamine or 1-o-alkyl-1-enyl-2-acyl-glyceroethanolamine. The major saturated fatty acids in phosphatidylcholine and phosphatidylethanolamine were C16:0 (30.3-67.5% and 23.2-54.5%) and C18:0 (3.6 17% and 15.4-28%), respectively, while the major unsaturated fatty acids in these lipids were C18:1n-9, n-7 (1.0-7.3% and 5.3-10.5%), C20:5n-3 (1.5-9.8% and 4,5 15.8%) and C22:6n-3 (12.5-42.0% and 7.0-11.3%), respectively. PMID- 10817216 TI - Antinociceptive properties of morusin, a prenylflavonoid isolated from Morus nigra root bark. AB - The antinociceptive effects of morusin (1), the main prenylflavonoid present in the Morus nigra root barks have been investigated in classical models of pain in mice. The results showed that 1 exhibits a promising antinociceptive or analgesic profile by the intraperitoneal route, being more potent than some standard drugs used as reference. The mechanism by which the morusin exerts antinociceptive activity still remains undetermined, but our results strongly suggest that it involves the participation of the opioid system. PMID- 10817217 TI - Transfer to in vitro conditions influences expression and intracellular distribution of galectin-3 in murine peritoneal macrophages. AB - Galectin-3 is a beta-galactoside-binding lectin that has been implicated in numerous physiological processes, including mRNA splicing, cell differentiation, tumor metastasis and the stress response. We have studied effects of transfer of resident murine peritoneal macrophages to in vitro conditions on galectin-3 in different cell compartments. Galectin-3 was purified by immunoprecipitation with rat monoclonal antibody M3/38, and analyzed by immunoblotting using the same antibody. Transfer to in vitro conditions nearly doubled the total amount of galectin-3 in cells, and caused significant alterations in its intracellular distribution, indicating that galectin-3 is involved in the adaptation of peritoneal macrophages to in vitro conditions. PMID- 10817218 TI - Effect of a pulsed magnetic field and of first cold-pressure sunflower oil on mice. AB - In previous studies it has been shown that exposure of mice to a 12-Hz 6 mT unipolar square pulsed magnetic field (PMF) suppressed the excess of weight due to application of 1st cold-pressure sunflower oil. This time we considered the effect of oil and/or PMF on the growing curves lifespans of mice. The exposure took place for 30 min 5 days a week, from the 7th week of life to death. The results are 1) a broken slope in the growing curves from the 125th day of aging: the exposed mice were lighter than the controls, keeping the differences between the growing curves needed a repeated exposure all life long; 2) a significant increase in the lifespan of the controls which received oil versus the controls which received water; 3) an increase in the lifespan of the exposed mice versus the non-exposed control batches. On one hand it has been reported that essential polyunsaturated fatty acids found in first cold-pressure sunflower oil played a prominent role in membrane structures and in immune equilibrium. On the other hand, it was shown that oscillating electric fields could activate Na+K+-ATPase. PMID- 10817219 TI - Protective effect of L-cysteine and glutathione on rat brain Na+,K+-ATPase inhibition induced by free radicals. AB - The aim of this study was to investigate whether the preincubation of brain homogenates with L-phenylalanine (Phe), L-cysteine (Cys) or reduced glutathione (GSH) could reverse the free radical effects on Na+,K+-ATPase activity. Two well established systems were used for the production of free radicals: 1) FeSO4 (84 microM) plus ascorbic acid (400 microM) and 2) FeSO4, ascorbic acid and H2O2 (1 mM) for 10 min at 37 degrees C in homogenates of adult rat whole brain. Changes in brain Na+,K+-ATPase activity and total antioxidant status (TAS) were studied in the presence of each system separately, with or without Phe, Cys or GSH. TAS value reflects the amount of free radicals and the capacity of the antioxidant enzymes to limit the free radicals in the homogenate. Na+,K+-ATPase was inhibited by 35-50% and TAS value was decreased by 50-60% by both systems of free radical production. The enzymatic inhibition was completely reversed and TAS value increased by 150-180% when brain homogenates were preincubated with 0.83 mM Cys or GSH. However, this Na+,K+-ATPase inhibition was not affected by 1.80 mM Phe, which produced a 45-50% increase in TAS value. It is suggested that the antioxidant action of Cys and GSH may be due to the binding of free radicals to sulfhydryl groups of the molecule, so that free radicals cannot induce Na+,K+ ATPase inhibition. Moreover, Cys and GSH could regulate towards normal values the neural excitability and metabolic energy production, which may be disturbed by free radical action on Na+,K+-ATPase. PMID- 10817220 TI - Site-specific in vivo targeting of magnetoliposomes using externally applied magnetic field. AB - Human serum albumin labeled with technetium-99m was encapsulated together with magnetite particles into phosphatidylcholine/cholesterol liposomes. In order to investigate the stability of this complex and its ability to be used for magnetic drug targeting, the in-vivo distribution after intravenous administration in rats was estimated. For in-vivo targeting an SmCo permanent magnet with intensity approximately 0.35 T was attached near the right kidney. Difference between the relative radioactivity in the magnetically targeted right kidney (25.92+/-5.84%) and non-targeted left kidney (0.93+/-0.05%) is sufficiently high for relevant clinical applications. PMID- 10817221 TI - A neural network model for the self-organization of cortical grating cells. AB - A neural network model with incremental Hebbian learning of afferent and lateral synaptic couplings is proposed,which simulates the activity-dependent self organization of grating cells in upper layers of striate cortex. These cells, found in areas V1 and V2 of the visual cortex of monkeys, respond vigorously and exclusively to bar gratings of a preferred orientation and periodicity. Response behavior to varying contrast and to an increasing number of bars in the grating show threshold and saturation effects. Their location with respect to the underlying orientation map and their nonlinear response behavior are investigated. The number of emerging grating cells is controlled in the model by the range and strength of the lateral coupling structure. PMID- 10817222 TI - Inhibitory effect of a cyclic urea derivative on rubella virus replication. AB - 1-(4-Morpholinomethyl)-tetrahydro-2(1H)-pyrimidinone (mopyridone) exhibited a marked activity against rubella virus (Judith and RA27/3 strains), a MIC50 value of 0.9 microM and selectivity ratio of 557.7 been found in the case of Judith strain. These data, in addition to the previous ones about its anti-alphavirus effects suggest the compound to be considered as a broad spectrum inhibitor of togavirus replication. PMID- 10817223 TI - Antinematodal activity of some tropical rainforest plants against the pinewood nematode, Bursaphelenchus xylophilus. AB - Sixty five methanolic extracts of Sumatran rainforest plants representing 63 species of 21 families were assayed in vivo for antinematodal activity against Bursaphelenchus xylophilus using our cotton ball-fungal mat method. Extracts of 27 plants species from 14 families exhibited antinematodal activity, while 37 species were inactive. Among them, three extracts of Bischofia javanica, Knema hookeriana and Areca catechu exhibited very strong activity at minimum effective dose (MED) of 0.7 mg/cotton ball (mg/bl.). Eight extracts from Allamanda cathartica, Ervatamia corymbosa, Hoya diversifolia, Bischofia javanica, Derris malacensis, Melastoma melabathricum, Ophiorriza konsteleary and Brucea sumatrana also showed strong activity (MED, 5 mg/bl.). PMID- 10817225 TI - Genotyping Cryptococcus neoformans var. neoformans with specific primers designed from PCR-fingerprinting bands sequenced using a modified PCR-based strategy. AB - Cryptococcus neoformans var. neoformans can be subdivided into six genotypes (VN1 VN6) based on different combinations of four major bands (420, 475, 540 and 800 bp) obtained by polymerase chain reaction (PCR) fingerprinting using the (GACA)4 primer. The aim of this study was to identify specific primers able to amplify these bands. A modified PCR-based sequencing strategy was adopted to overcome the limitations of using (GACA)4 as a single cycle sequencing primer. The original bands, made up of amplicons with two terminal (GACA)4 sequences, were digested with five restriction enzymes. Digestion products yielding two or three fragments were selected. Each fragment was expected to have no more than one terminal (GACA)4 sequence, making cycle sequencing possible. Fragments were purified and sequenced with the (GACA)4 primer. New primers specific for each of the four major bands were then designed and the remaining regions were sequenced using both purified bands and PCR-fingerprinting products as template. These primers were used to amplify the genomic DNA of 12 C. neoformans strains and five strains of other yeast species. The new primers, used as separate pairs or in a mixture of all pairs, amplified the expected bands only in C. neoformans var. neoformans strains, confirming the species specificity of the bands selected for molecular typing of this yeast. PMID- 10817224 TI - Two antinematodal phenolics from Knema hookeriana, a Sumatran rainforest plant. AB - The activity-guided chromatographic purification of the methanol extract of Knema hookeriana, using pine wood nematodes Bursaphelenchus xylophilus has successfully led to the isolation and characterization of two phenolic antinematodal compounds with minimum effective dose (MED) of 4.5 and 20 microg/cotton ball (microg/bl.) or 0.018 and 0.073 microM/cotton ball (microM/bl.), respectively. Based on their chemical and spectral properties, these compounds were determined to be 3 undecylphenol (1) and 3-(8Z-tridecenyl)-phenol (2). These compounds were isolated for the first time from this species, and 2 seems to be a novel compound. PMID- 10817226 TI - Precipitate production by some Malassezia species on Dixon's agar. AB - The production of a precipitate by Malassezia species on Dixon's agar was observed. Malassezia furfur (n = 12), M. obtusa (n = 2) and M. slooffiae (n = 3) were precipitate negative, while M. sympodialis (n = 32) and M. globosa (n = 6) were precipitate positive. This test may be useful in differentiating Malassezia species. PMID- 10817227 TI - Presence of a pertussis toxin-sensitive G protein alpha subunit in Sporothrix schenckii. AB - As an initial step in the study of the role of G proteins in signal transduction in Sporothrix schenckii, we identified a Galphai subunit using different experimental approaches. Western blots of fungal membrane preparations using anti Galphacommon and anti-Galphai1-Galphai2 antibodies identified a band of approximately 41 kDa. Pertussis toxin-catalyzed adenosine diphosphate (ADP) ribosylation of these membrane fractions confirmed the presence of a protein substrate of 41 kDa. A 357 bp polymerase chain reaction (PCR) product obtained using fungal DNA as template and primers targeted to conserved Galphai sequences, was used as a probe to isolate a clone from an S. schenckii genomic library. A partial sequence for a Galphai subunit was obtained from this clone. The sequence was completed using the rapid amplification of cDNA ends (RACE) technique with mycelium and yeast cDNA. The cDNA sequence revealed a 1059 bp open reading frame encoding a 353 amino acid Galphai subunit of 41 kDa, more than 90% identical to the CPG-1 of Cryphonectria parasitica, and GNA-1 of Neurospora crassa. The genomic sequence was obtained by PCR using fungal DNA, and revealed a 1250 bp sequence and the presence of three introns. These results provide evidence for the first time of the presence and expression of a Galphai homolog in a pathogenic dimorphic fungus. PMID- 10817228 TI - Identification of Candida albicans clinical isolates by PCR amplification of an EFB1 gene fragment containing an intron-interrupted open reading frame. AB - The use of a single pair of primers, deduced from the intron and exon nucleotide sequences of the Candida albicans EFB1 gene, in polymerase chain reaction (PCR) assays performed with whole cells of both laboratory strains and clinical isolates of Candida species, resulted in the species-specific amplification of a 785 bp DNA fragment in C. albicans strains. Clinical C. albicans isolates were tested, and 85 out of 86 generated the expected PCR-amplified product; other Candida species, both laboratory strains and clinical isolates, as well as laboratory strains belonging to other fungal genera, including medically relevant taxa, failed to amplify any DNA fragment. In addition, unusual C. albicans isolates (glucosamine- and N-acetylglucosamine-negative) from Africa also yielded the expected PCR-generated DNA fragment. These results indicate that genes containing intron sequences may be useful to design species-specific primers for the identification of fungal strains by PCR. PMID- 10817229 TI - Flow cytometric assessment of human peripheral blood mononuclear cells in response to a coccidioidal antigen. AB - The in vitro responses of peripheral blood mononuclear cells (PBMC) from healthy immune and non-immune donors were assessed by flow cytometry after incubation with the coccidioidal antigen toluene spherule lysate (TSL). After 120 h of incubation with 100 microg ml(-1) of TSL, expression of the activation markers CD69, CD25 and human leukocyte antigen-DR were all significantly increased in CD3+ lymphocytes from immune donors compared to non-immune donors (P < 0.03 for all). No differences in the surface expression of the costimulatory molecules CD28, CD152 or CD154 was seen between immune and non-immune donors after either 24 or 120 h of TSL incubation, nor were differences detected in the expression of the B7 ligands CD80 or CD86 on CD14+ monocytes. The percent of CD3+ lymphocytes expressing intracellular interferon-gamma (IFN-gamma) was significantly increased in immune compared to non-immune donors and was further increased by the addition of 10 ng ml(-1) of human recombinant interleukin (IL)-12 (P < 0.05 for both). Both CD4+ and CD8+ lymphocytes contributed to IFN-gamma production. These data indicate that coccidioidal antigen stimulation of lymphocytes from healthy immune donors leads to specific expression of activation molecules and production of intracellular IFN-gamma. Addition of IL-12 leads to a significant recruitment of cells producing IFN-gamma among immune donors. PMID- 10817230 TI - The effects of diffusates from the spores of Aspergillus fumigatus and A. terreus on human neutrophils, Naegleria gruberi and Acanthamoeba castellanii. AB - Diffusates from dormant spores of Aspergillus fumigatus are inhibitory to certain functions of immune phagocytic cells and soil protozoa. An assay of human neutrophil phagocytosis and an in vitro method of measuring inhibition of the growth of free living amoebae are described. The anti-phagocytic and anti-amoebal effects of diffusates from clinical and environmental isolates of A. fumigatus and of A. terreus were measured using these methods. Spore diffusates from all isolates of A. fumigatus and A. terreus tested significantly inhibited neutrophil phagocytosis. Spore diffusates from A. fumigatus, but not A. terreus, significantly inhibited the growth of Naegleria gruberi. Spore diffusate from A. fumigatus did not inhibit the growth of Acanthamoeba castellanii. The relevance of these findings to the pathogenicity of A. fumigatus and A. terreus is discussed. PMID- 10817231 TI - Molecular and conventional taxonomy of the Microsporum canis complex. AB - The validity of taxa around Microsporum canis was evaluated by a combination of phenetic and molecular methods. Morphological and physiological features were compared with results of sequencing of the internal transcribed spacer (ITS) region of the ribosomal operon, polymerase chain reaction (PCR) fingerprinting and amplified fragment length polymorphism (AFLP) analysis. The seven species investigated seem to be infraspecific taxa and were reclassified or synonymized as M. canis (teleomorph: Arthroderma otae), M. ferrugineum, and M. audouinii. PMID- 10817232 TI - An evaluation of the in vitro activity of terbinafine. AB - Terbinafine has previously been shown to be highly active against dermatophytes and many other filamentous fungi. However, its activity against yeasts is controversial, with earlier reports suggesting that it has low activity, while more recent studies demonstrated that terbinafine is effective against yeasts. In this study, the in vitro activity of terbinafine was evaluated against a broad range of fungal isolates. We examined the susceptibility of 100 yeast strains (10 species including Candida albicans, non-C. albicans, fluconazole-susceptible and resistant candidal strains), and 184 strains of filamentous fungi and dermatophytes (29 species including Aspergillus, Fusarium, Sporothrix, Trichophyton rubrum, T. mentagrophytes, T. tonsurans, Microsporum canis and Epidermophyton floccosum), using the NCCLS M27-A microdilution methodology for yeasts and a modified M38-P methodology for moulds. The endpoint for terbinafine was defined as 80% inhibition compared with the growth control well. The mean yeast and filamentous fungi minimum inhibitory concentration values +/- SEM (in microg ml(-1)) for terbinafine were: 6.60 +/- 0.73 and 1.04 +/- 0.28, respectively. In conclusion, our data suggest that terbinafine, in addition to its potent activity against dermatophytes, is considerably effective against a broad range of yeasts and filamentous fungi in vitro. Therefore, investigations concerning its antifungal activity in vivo against such organisms should be pursued. PMID- 10817233 TI - Antifungal activity of splenic, liver and pulmonary macrophages against Candida albicans and effects of macrophage colony-stimulating factor. AB - Disseminated infections due to Candida albicans are frequently encountered in immunocompromised patients. We compared the antifungal activities of macrophages residing in spleen, liver and lungs of rabbits against blastoconidia and pseudohyphae of C. albicans. Splenic adherent cells (SAC), Kupffer cells (KC) and pulmonary alveolar macrophages (PAM) all ingested blastoconidia efficiently. SAC caused significantly more damage to unopsonized pseudohyphae compared with KC (P < 0.01) or PAM (P < 0.001). Incubation of SAC with 15 ng ml(-1) of recombinant human macrophage colony-stimulating factor (M-CSF) at 37 degrees C for 2 days significantly enhanced phagocytosis (P = 0.02) and killing (P = 0.05) of blastoconidia. In contrast, M-CSF had no effect on phagocytic activities of KC or PAM against blastoconidia or on damage caused by any of the macrophages to pseudohyphae of C. albicans. Thus, although all three resident macrophage types ingest blastoconidia efficiently, they differ in their capacity to cause damage to pseudohyphae and in their responsiveness to M-CSF for antifungal activation. M CSF augments the capacity of SAC to ingest and kill blastoconidia and may therefore have a role in the treatment and prevention of hematogenously disseminated candidiasis. PMID- 10817234 TI - Subcutaneous phaeohyphomycosis in transplant recipients: review of the literature and demonstration of in vitro synergy between antifungal agents. AB - Infection of subcutaneous tissue by black fungi (subcutaneous phaeohyphomycosis) has only been reported in six transplant patients, all of whom were solid organ recipients. These patients presented with indolent, localized infections at least 1 year after transplant, while on maintenance immunosuppressive regimens. They were cured by surgical resection, either alone or in conjunction with antifungal agents. We report a case of subcutaneous phaeohyphomycosis occurring in a bone marrow transplant recipient receiving high doses of immunosuppressive agents, in whom widespread subcutaneous infection due to Exophiala jeanselmei was not eradicated by repeated resections and therapy with amphotericin B and flucytosine. The infection was eventually cured after addition of itraconazole to the therapeutic regimen. Results of in vitro testing of the isolate for susceptibility to a combination of amphotericin B, flucytosine and itraconazole confirmed the potential role of combination antifungal therapy in the setting of refractory infection. PMID- 10817235 TI - Effect of cytokines on antifungal activity of human polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis. AB - In our previous study, it was observed that human peripheral blood polymorphonuclear leucocytes (PMNs) exhibited a fungistatic effect on yeast cells of Paracoccidioides brasiliensis, and that interferon-gamma (IFN-gamma), but not tumor necrosis factor-alpha or interleukin-8 (IL-8), enhanced the antifungal activity of PMNs. In the present study, granulocyte-macrophage colony-stimulating factor (GM-CSF) also enhanced the PMN activity. GM-CSF-activated PMNs exhibited a killing effect on P. brasiliensis isolate Bt-4 and an enhanced fungistatic effect on isolate Aoki. IL-1beta activated PMNs to kill isolate Bt-4. Granulocyte colony stimulating factor had no effect. Combinations of IFN-gamma with GM-CSF or IL 1beta, but not a combination of GM-CSF and IL-1beta, exhibited a synergistic effect in enhancing the antifungal activity of PMNs. These results strongly suggest that PMNs activated with IFN-gamma, GM-CSF and/or IL-1beta might play an important role in host defense in early infection with P. brasiliensis due to their enhanced antifungal activity. PMID- 10817236 TI - Accreditation and point-of-care testing. PMID- 10817237 TI - Measurement of leptin and leptin binding in the human circulation. PMID- 10817238 TI - Measurement of immune complexes is not useful in routine clinical practice. PMID- 10817239 TI - Aldosterone and renin measurements. PMID- 10817240 TI - Role of biochemical assessment in management of corticosteroid withdrawal. PMID- 10817241 TI - Effect of the inflammatory response on trace element and vitamin status. PMID- 10817242 TI - Transient pituitary hypothyroidism in a patient with ectopic adrenocorticotrophic hormone secretion. AB - We report the case of a 55-year-old woman who presented with hypercortisolism secondary to ectopic adrenocorticotrophic hormone secretion and severe non thyroidal illness syndrome (NTIS) due to metastatic small cell lung carcinoma associated with severe infections. The patient initially showed hormonal profiles of pituitary hypothyroidism and gonadal hypofunction. After decrease in cortisol production following treatment with chemotherapy and metyrapone, serum thyroid hormones and thyroid-stimulating hormone (TSH) concentrations normalized. Study of the relative contributions of cortisol and pro-inflammatory cytokines (interleukin-6 and tumour necrosis factor alpha) to the overall variability in thyroid function tests disclosed a significant and independent effect of serum cortisol on serum TSH concentrations; the variability in free thyroid hormone concentration was explained only by changes in TSH concentration. These observations indicate that cortisol could be the major determinant of changes in serum TSH concentrations in clinical conditions accompanied by hypercortisolism, as occurs in NTIS. PMID- 10817243 TI - Occurrence of hyperhomocysteinaemia in cardiovascular, haematology and nephrology patients: contribution of folate deficiency. AB - We investigated the contribution of plasma folate deficiency to hyperhomocysteinaemia in selected patient groups. Based on our observations, we have determined a lower folate reference interval cut-off using homocysteine as a metabolic marker of folate deficiency. Four hundred and twenty-five consecutive plasma specimens from cardiology (n = 120), haematology (n = 190) and nephrology (n = 115) patients were analysed for homocysteine and plasma folate concentrations. Healthy volunteers were used as controls (n = 117). We observed elevated homocysteine values above our upper reference limit of 13 micromol/L in 20.1%, 28.4% and 74.8% of the cardiology, haematology and nephrology patients, respectively. All but 1.9% of the patients had plasma folate values greater than the lower reference interval limit (3.4 nmol/L) for our folate assay. The percentage of patients from cardiology and haematology clinics who were hyperhomocysteinaemic and had folate values > 15 nmol/L was 5.0% and 4.2%, respectively. In contrast, 58% of our nephrology patients with folate values > 15 nmol/L were hyperhomocysteinaemic. In all three groups, an inverse relationship was found between folate and homocysteine. The folate/homocysteine ratios in the patient groups were approximately one-third of the values observed in our control group. Folate deficiency appears to be the primary cause of hyperhomocysteinaemia in our cardiology and thrombosis patients. However, severe folate deficiency appears to be uncommon. The majority of our nephrology patients are hyperhomocysteinaemic without an apparent folate deficiency. We conclude that raising the lower reference interval cut-off for folate to 15 nmol/L would help to identify individuals at risk for hyperhomocysteinaemia in our non-uraemic patient population. Increasing folate supplementation to maintain a plasma concentration above 15 nmol/L in cardiac, thrombosis and renal patients would greatly reduce the occurrence of hyperhomocysteinaemia in these patients. PMID- 10817244 TI - LDL subfractions in patients with myocardial infarction: effect of smoking and beta-blocker treatment. AB - The aim of the present case-control study was to estimate, by density gradient ultracentrifugation, LDL heterogeneity in myocardial infarction, and to evaluate the effect of smoking and beta-blocker treatment on LDL subfraction profile. Our results show that patients who survive myocardial infarction have an abundance of small, dense LDL in their plasma, compared with controls. Patients who were on beta-blockers and those who smoked showed a more atherogenic LDL subfraction profile than the rest. In patients on beta-blocker treatment, the proportion of LDL3 was positively correlated with triglyceride concentration and body mass index. Dense LDL predominates in patients irrespective of smoking or beta-blocker treatment. The relative risk, calculated by logistic regression as the odds ratio of high LDL3, was 7.5 (95% confidence interval 2.5-22.1) and was not significantly influenced when smoking, beta-blocker treatment, triglycerides or the other parameters of the study were included in the statistical model. PMID- 10817245 TI - Radioimmunoassay for the measurement of thrombospondin in plasma and breast cyst fluid: validation and clinical application. AB - Thrombospondin is an adhesive protein that has been implicated in malignancy, specifically in tumour progression and angiogenesis. We developed a radioimmunoassay for the measurement of thrombospondin in plasma and breast cyst fluid. The assay exhibited high accuracy, with recoveries of 102-136% and acceptable imprecision, with an intra-assay coefficient of variation (CV) of <7.5% across the analytical range 30-1000 ng/mL and inter-assay CV of 4.4% and 7.7% at 152 and 224 ng/mL, respectively. Thrombospondin measured in the breast cyst fluid of patients with gross cystic disease of the breast showed that patients with type II (Na+) cysts had significantly higher concentrations than type I (K+) cysts. The plasma thrombospondin reference range was determined as 131-274 ng/mL. Patients with breast cancer had significantly higher plasma thrombospondin concentrations than normal individuals or patients with benign breast disease. Plasma thrombospondin was higher in breast cancer patients with lymph node involvement. PMID- 10817246 TI - Evaluation of assays of serum IgG subclasses and IgG antigen-specific antibodies in the investigation of recurrent infection. AB - Patients who have recurrent infections require laboratory investigation for possible underlying immunodeficiency. We retrospectively evaluated the usefulness of two relevant assays (serum IgG subclass concentration and levels of IgG antigen-specific antibodies to a panel of relevant carbohydrate and protein antigens) in the management of patients referred to a regional clinical immunology service over a 3-year period. Of 97 patients for whom both assays were performed, five (5.2%) had a low result for IgG subclasses but this did not influence the management of any patient; 51 patients (53%) had a low result for antigen-specific antibodies and this influenced management in 43 cases. We conclude that knowledge of the serum IgG subclass concentrations is of dubious relevance and this calls into question whether the assay should continue to be offered. This regional service has ceased to perform the assay routinely in such patients. Our findings require confirmation by a larger multicentre study that should assess the clinical outcome of any changes in practice. PMID- 10817247 TI - A model for harmonization of routine clinical chemistry results between clinical laboratories. AB - Clinical chemistry laboratory results from different laboratories often show large between-laboratory variation due to factors such as differences in method principles, method applications, calibration procedures or the application of different instrument factor settings within the same calibration procedure. We have examined the possible use of common calibrators to reduce this variation. Three different calibrators were compared: A, freeze-dried preparations of pooled patients' serum samples, spiked to give three concentration levels; B, freeze dried preparations of pooled patients' serum samples selected on the basis of elevated enzyme activities at three levels; C, a single calibrator consisting of frozen pooled serum samples. These calibrators were sent to 11 participating laboratories together with 14 fresh patients' serum samples. We report the variation of the results of 21 general clinical chemistry analytes obtained in the patients' serum samples before and after recalculation on the basis of the results of the calibrators. For most analytes the use of a multiple point linear regression calibration function is able to reduce the between-laboratory variation considerably from more than 30% (enzymes) to values well within the bias limits set by European quality specifications, when the necessary conditions are met. These conditions include the commutability of the calibrator(s) with fresh patients' material. For the enzymes, calibrator material originating from selectively pooled patients' samples appeared to be necessary, whereas for the substrates selectively pooled serum calibrators spiked with exogenous supplements may be used. For harmonization to be effective in practice, calibrators need to be stable over time and to carry assigned values set by certified reference laboratories, and the quality performance of participating laboratories should be appropriately monitored. PMID- 10817248 TI - Simple high-performance liquid chromatographic method to monitor vigabatrin, and preliminary review of concentrations determined in epileptic patients. AB - A simple and rapid high-performance liquid chromatographic method has been developed for the determination of vigabatrin concentrations in plasma or serum. The assay uses only 100 microL of specimen and has been found to be linear over a concentration range of 1 to 50 mg/L. The limit of detection has been determined as 1 mg/L, and the between-batch coefficient of variation for the two internal quality controls routinely analysed (n = 33) has been found to be less than 5%. There was no evidence of interferences in the assay from other commonly prescribed anti-epileptic drugs. This method has been applied to routine clinical specimens to determine the concentration of vigabatrin in 47 patient specimens over a 12-month period. It was found that only 63% of the male group and 53% of the female group were within the proposed target concentration of 5 to 35 mg/L. In addition, it was found that 26% of the male group and 36% of the female group were found to have concentrations below 5 mg/L, which may indicate lack of compliance and/or lack of therapeutic efficacy of treatment. PMID- 10817249 TI - New method for faecal fat determination by mid-infrared spectroscopy, using a transmission cell: an improvement in standardization. AB - Current techniques used in clinical laboratories for faecal fat determination, such as the Van de Kamer method, are not very accurate or precise. This became apparent when results obtained by different laboratories were compared, and could explain the disappointing performance of near-infrared and mid-infrared spectroscopy since the accuracy of these techniques depends upon the accuracy of the calibration used (i.e. inaccurate wet chemical analysis). In order to improve standardization, we developed and tested a new quantitative method in three laboratories, based on Fourier transform infrared (FT-IR) spectroscopy. Fatty acids were extracted from faecal samples with acidified petroleum ether-ethanol and the extracts were dried and dissolved in chloroform. An infrared spectrum of the extracts was recorded in the range 4000-650 cm(-1), using an infrared transmission cell. Standard mixtures of stearic and palmitic acids (65:35) were used for calibration. Quantification was based on the absorbance band of the CH2 group (2855 cm(-1)) of free fatty acids and fatty acid glycerol esters. The calibration curve showed excellent linearity. The correlation coefficient between the titrimetric Van de Kamer and FT-IR methods was 0.96 (y = 1.12x-0.02, standard error of prediction = 0.89 g% fat). No significant difference was found when the FT-IR results of 28 faecal samples from patients were compared between two different university hospital laboratories. The new FT-IR method, using primary standards, is simple and rapid, and provides satisfactory intra- and inter laboratory precision for the diagnosis and monitoring of steatorrhoea. PMID- 10817250 TI - Application and validation of a urinary methadone metabolite (EDDP) immunoassay to monitor methadone compliance. AB - A total of 1381 urine specimens were screened using a Microgenics CEDIA urinary primary methadone metabolite (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine; EDDP) immunoassay (cut-off calibrator concentration of 100 microg/L) in combination with a Dade Behring EMIT urinary methadone immunoassay (cut-off calibrator concentration of 300 microg/L). Of these, 642 (46%) were found to be positive using the EDDP assay but only 541 (39%) were found to be positive by the methadone assay. Out of the 108 specimens which were EDDP-positive but negative by the methadone assay, 47 (7%) could not be confirmed as positive using the routine in-house method of gas chromatography incorporating nitrogen-specific detection. Of these 47 results, 38 were re-screened using a more sensitive gas chromatography-mass spectrometry (GC-MS) technique, which demonstrated the presence of EDDP in every case. There was insufficient specimen to analyse the remaining nine samples. There were seven specimens which gave negative results by the EDDP and GC-MS assays but which had methadone concentrations ranging from 4000 to 37,500 microg/L. These were therefore presumed to be 'spiked' with methadone, i.e. to have had methadone added to the specimen to yield positive screening results and simulate compliance. It is concluded that the Microgenics CEDIA EDDP assay is a sensitive and reliable technique to determine the compliance of subjects prescribed methadone for opiate detoxification and maintenance. PMID- 10817251 TI - Effect of urine storage on urinary uric acid concentrations. AB - Accurate determination of serum and urinary uric acid concentrations is essential for the diagnosis and classification of gout according to uric acid metabolism derangement. Urine and/or serum samples are often kept at either 4 degrees C or 20 degrees C until assayed, when a large number of samples are handled simultaneously. Our preliminary study indicated a significant decrease in urinary uric acid concentration after preservation, regardless of the storage temperature. Uric acid crystals were often observed in these cases which showed a marked decrease in urinary uric acid concentration after storage. In the present study, we sought the factor(s) that might cause this decrease in urinary uric acid concentration, as well as measures to overcome the problem. High urinary uric acid concentration and low pH proved to play major roles in the decrease in urinary uric acid concentration after storage. In contrast, dilution of the urine samples before storage resulted in no significant change in urinary uric acid concentration. Based on these results, we recommend diluting urine before storage for determination of uric acid concentration and avoiding underestimation. PMID- 10817252 TI - Apo(a) isoforms do not predict risk for coronary heart disease in a Gulf Arab population. AB - It is well recognized that blood lipoprotein A [Lp(a)] levels constitute an important risk factor for atherosclerotic vascular disease. In some populations, mainly Caucasian, Lp(a) levels and coronary heart disease (CHD) risk are determined by the pattern of apolipoprotein a [apo(a)] polymorphism. It is currently unclear if these observations apply to other populations and ethnic groups. The aim of the current study is to determine to what extent known apo(a) polymorphisms associate with development of CHD in a Kuwaiti Arab population. Serum Lp(a) levels were measured by enzyme-linked immunosorbent assay and apo(a) isoforms determined by a high-resolution sodium dodecyl sulphate/agarose gel electrophoresis with immunoblotting in two groups of Kuwaiti subjects: healthy controls (n = 140) and subjects with CHD (n = 140). Blood lipids and anthropometric parameters were also determined in these subjects by standard methods. Serum Lp(a) levels were greater in those with CHD than in those in the healthy group (P < 0.001). There was no consistent trend in the pattern of serum Lp(a) levels found with specific apo(a) isoforms in either group of subjects. There was, therefore, no simple relationship between the isoform pattern (and number of kringle-IV repeats) and serum Lp(a) concentration, unlike in certain other populations. Additionally, almost identical proportions of subjects in either group had single-banded (homozygous, approximately 70%), double-banded (heterozygous, approximately 23%) and no-band (null, approximately 7%) phenotypes. The distribution of the five identified isoforms (F, S1, S2, S3 and S4) also was almost identical for both groups of subjects, whether homozygous or heterozygous, and whether classified into fast-moving (F, S1 and S2) or slow moving (S3 and S4) isoforms. We conclude that the frequency and pattern of distribution of apo(a) phenotypes did not differ significantly between healthy control Kuwaiti Arab subjects and those with CHD. It is thus unlikely that an individual's apo(a) phenotype can predict both serum Lp(a) level and risk for CHD, irrespective of race and/or ethnic grouping. PMID- 10817253 TI - Hyperproinsulinaemia in elderly subjects: evidence for age-related pancreatic beta-cell dysfunction. AB - We investigated whether pancreatic beta-cell dysfunction has a role in the pathogenesis of glucose intolerance in the elderly by comparing plasma glucose, immunoreactive insulin, C-peptide and proinsulin concentrations during a 100-g oral glucose load in six healthy older male volunteers [aged 69 (65.9-74.9) years; median (95% confidence limits)] and seven young male controls [aged 24.0 (21.9-26.3) years]. Fasting and integrated concentrations of glucose and C peptide were similar in the elderly and young. Although fasting, insulin and proinsulin levels were similar, integrated insulin (P=0.05) and proinsulin (P<0.05) concentrations were higher in the elderly than in controls. Insulin resistance, measured as homeostasis model assessment, was greater (P<0.05) in the elderly than in controls. Elderly men had greater molar ratios of integrated insulin:C-peptide (P<0.05) and proinsulin:C-peptide (P<0.01) but their respective fasting molar ratios were similar when compared with controls. Pancreatic beta cell secretion in older subjects, as assessed by C-peptide concentrations, was inappropriately low in the presence of insulin resistance. Their post-prandial 'hyperinsulinaemia' is probably due to a combination of hyperproinsulinaemia and reduced metabolic clearance of insulin. Older subjects had disproportionately high proinsulin to C-peptide levels, suggesting pancreatic beta-cell dysfunction. These results are consistent with the notion that decreased insulin sensitivity and pancreatic beta-cell dysfunction may predispose the elderly to glucose intolerance. PMID- 10817254 TI - A reliable non-separation fluorescence quenching assay for total glycated serum protein: a simple alternative to nitroblue tetrazolium reduction. AB - A simple non-separation assay for the measurement of total glycated serum protein is described. It was found that the fluorescence intensity of a solution of a fluorescein-boronic acid derivative was quenched in proportion to the amount of serum added. This led to the development of an assay in which 10 microL of serum is added to 4 mL of a solution of the fluorescein-boronic acid derivative and the fluorescence intensity is measured after 15 min. The results, as measured by drop in fluorescence intensity, calibrated by a single standard, were compared with the results for nitroblue tetrazolium (NBT) reduction of fructosamine and showed good correlation (r=0.936, n=114). The intra-assay precision (seven samples each measured 10 times) was less than 2.1% (concentration range 190-660 micromol/L); inter-assay precision for seven samples in 10 assays was less than 2.5% (over the same concentration range). Dilution of serum that had a high concentration of total glycated protein showed the assay to be linear. Serum samples (with low, medium and high total glycated protein concentrations) showed less than 2.1% difference from base results with added glucose (up to 60 mmol/L), less than 9.7% difference with added bilirubin (up to 250 micromol/L) and less than 6.9% with added triglycerides (up to 50 mmol/L). Addition of haemoglobin (up to 0.9 g/dL) with high glycation (11.7% HbA1c) to plasma (298 micromol/L total glycated protein) showed less than 10% difference from the base result. Assays performed over a range of temperatures (12-34 degrees C) showed no significant differences in the results. The assay gives similar results to the currently used NTB method but with significantly less susceptibility to interferences. As such the method should be a useful aid in the management of diabetes. PMID- 10817255 TI - Quantitative and qualitative analysis of haemoglobin variants using capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE) has been introduced into the clinical chemistry laboratory because of its range of potential applications. In this paper, we evaluate an alkaline CZE method for the quantification of HbA2 and HbF and also assess the combination of the alkaline CZE method with an acid CZE method for the determination of haemoglobin variants in an automated fashion. Correlation of the HbA2% determined between the HbA2-CZE method (alkaline conditions) and the Helena Sickle Thal Quick Column method was good (r = 0.91). The correlation between the HbF% determined by the HbA2-CZE method and by the alkaline denaturation method was acceptable (r = 0.81). The HbA2-CZE method was able to identify a large number of haemoglobin variants. The variants HbC and HbE or HbS and HbD, however, had the same characteristics under alkaline conditions and could therefore not be discriminated from each other. The identification of these overlapping variants could be accomplished by the analysis of the blood specimens in combination with the HbA1c-CZE method (acid conditions). We conclude that the presented applications for capillary zone electrophoresis can be used for quantitative and qualitative analysis of haemoglobin variants. PMID- 10817256 TI - Non-fasting reference intervals for the Abbott IMx homocysteine and AxSYM plasma folate assays: influence of the methylenetetrahydrofolate reductase 677 C-->T mutation on homocysteine. AB - Plasma homocysteine comes under both genetic and nutritional control. B vitamins and particularly folate are important factors in homocysteine metabolism. We have obtained reference intervals for total plasma homocysteine and plasma folate. We have also determined the influence of methylenetetrahydrofolate reductase (MTHFR) genotype on plasma homocysteine concentrations in healthy individuals. Reference intervals for Abbott IMx homocysteine and AxSYM plasma folate assays were established using 116 volunteers recruited from hospital staff. Exclusion criteria included cardiac, hepatic or renal disorders, and use of over-the counter prescription medications. An exception was the inclusion of three women using oral contraceptives and one woman receiving post-menopausal oestrogen supplementation. Methylenetetrahydrofolate reductase 677C-->T genotyping was performed on 101 of the volunteers to determine whether the MTHFR 677T allele influences homocysteine concentrations in healthy individuals. Reference intervals for homocysteine and folate were determined using the mean+/-2 standard deviations of the data. Folate/homocysteine ratios were sorted by MTHFR C677T genotype. Homocysteine correlated negatively with plasma folate. Mean male homocysteine concentrations were significantly higher (9.0 micromol/L; P<0.05) than the mean value (7.1 micromol/L) obtained for females. Mean homocysteine values were significantly higher in subjects who were homozygous for the MTHFR 677T allele when compared with the 677CC genotype (P<0.05). Ratios of folate/homocysteine were 20% and 7.4% lower in the male and female 677TT group than in the 677CC group, respectively. The mean homocysteine value of 43 volunteers who were taking multivitamins was not significantly different from that of 73 who were not vitamin supplemented. Conversely, the mean folate value was slightly greater, and statistically significant, in the group taking vitamin supplements. The mean folate values and reference intervals were not significantly different when grouped by sex or age. MTHFR 677C-->T mutations influenced homocysteine values observed in our study of healthy volunteers, even though we did not observe outright folate-deficient individuals. Our random homocysteine values were similar to the fasting homocysteine values obtained in other studies. PMID- 10817257 TI - Micro-flowcell conductometric sweat analysis for cystic fibrosis diagnosis. AB - This paper describes a device specifically designed to facilitate neonatal sweat testing. The components are sized appropriately for attachment to the limbs of newborns. Iontophoretic electrodes, with pilocarpine gel inserts, are latched into small holders attached by straps to the limb. The holder at the anodic site remains in place to receive and align the sensor cell, which uses a conical collecting surface to channel the sweat directly and anaerobically from the sweat ducts to the continuous flow-through conductivity cell within its body. A crib side analysis unit incorporates an iontophoretic power supply and displays a continuous readout of sweat electrical conductivity. The average conductivity during a specific time interval and the initial sweating rate are automatically displayed. The method, which simplifies sweat tests, is currently being assessed in three neonatal clinical trials to test its ability to reduce test failures in the newborn due to insufficient sweat. PMID- 10817258 TI - Bilirubin interference with a salicylate assay performed on an Olympus analyser. PMID- 10817259 TI - Falsely low vancomycin results using the Abbott TDx. PMID- 10817260 TI - Human bone cell cultures in biocompatibility testing. Part I: osteoblastic differentiation of serially passaged human bone marrow cells cultured in alpha MEM and in DMEM. AB - Well-characterised human osteoblastic bone marrow cell cultures are a useful in vitro tool to analyse bone tissue/biomaterials interactions. In this work, human bone marrow was cultured in experimental conditions described to favour osteoblastic differentiation and, serially passaged cells were cultured in two widely used culture media, minimum essential medium Eagle, alpha modification (alpha-MEM) and Dulbecco's modified Eagle's medium (DMEM). Cultures were grown for 35 d and compared concerning morphologic appearance on scanning electron microscopy (SEM), cell viability/proliferation, total protein content, activity of alkaline phosphatase (ALP) and ability to form calcium phosphate deposits. Results showed that cell proliferation was similar in cultures grown in the two media but ALP activity and ability to form mineralised deposits were lower in DMEM cultures. In both experimental situations, osteoblastic parameters were strongly reduced on cell passage, particularly from the first to the second subculture. In the experimental conditions used (presence of ascorbic acid, sodium beta-glycerophosphate and dexamethasone in the primary and secondary cultures), osteoblastic differentiation was observed in the first and second subcultures grown in alpha-MEM and in the first subculture grown in DMEM. These results underline the importance of the definition of the experimental conditions in studies involving bone cell cultures. PMID- 10817261 TI - Human bone cell cultures in biocompatibility testing. Part II: effect of ascorbic acid, beta-glycerophosphate and dexamethasone on osteoblastic differentiation. AB - This work analyses the proliferation/differentiation behaviour of human bone marrow cells cultured in alpha-minimum essential medium supplemented with 10% foetal bovine serum (standard medium) and in the presence of ascorbic acid (AA, 50 microg ml(-1)), beta-glycerophosphate (betaGP, 10 mmol) and dexamethasone (Dex, 10 nmol) under selected experimental conditions. Cultures were compared concerning cell morphology, cell growth, ALP activity and ability to form calcium phosphate deposits. Cells growing in the various experimental conditions proliferated gradually with the incubation time and presented high ALP activity. Cultures grown in standard medium and in the presence of either AA or Dex failed to form calcium phosphate deposits. Cultures grown in the presence of betaGP, betaGP + AA and betaGP + AA + Dex, i.e. in the presence of a source of phosphate ions, showed the formation of a mineralised extracellular matrix. The presence of Dex resulted in a significant induction in the ALP activity and ability to form mineral deposits. The behaviour of the various cell cultures is in agreement with previous studies stating a reciprocal and functionally coupled relationship between proliferation and differentiation, i.e. cultures grown in a medium containing betaGP presented a less proliferative but more differentiated osteoblastic cell population, as compared to cultures lacking the mineralisation process. PMID- 10817262 TI - The effects of calcium phosphate cement particles on osteoblast functions. AB - Calcium phosphate cements (CPC) are increasingly used in the orthopedic field. This kind of cement has potential applications in bone defect replacements, osteosynthetic screw reinforcements or drug delivery. In vivo studies have demonstrated a good osteointegration of CPC. However, it was also observed that the resorption of CPC could create particles. It is known from orthopedic implant studies that particles can be responsible for the peri-implant osteolysis. Biocompatibility assessment of CPC should then be performed with particles. In this study, we quantified the functions of osteoblasts in the presence of beta TCP, brushite and cement particles. Two particle sizes were prepared. The first one corresponded to the critical diameter range 1-10 microm and the second one had a diameter larger than 10 microm. We found that CPC particles could adversely affect the osteoblast functions. A decrease in viability, proliferation and production of extracellular matrix was measured. A dose effect was also observed. A ratio of 50 CPC particles per osteoblast could be considered as the maximum number of particles supported by an osteoblast. The smaller particles had stronger negative effects on osteoblast functions than the larger ones. Future CPC development should minimize the generation of particles smaller than 10 microm. PMID- 10817263 TI - Drug release behavior of beads and microgranules of chitosan. AB - Beads and microgranules carriers have important potential applications for the administration of therapeutic molecules. A novel approach for the preparation of chitosan beads and microgranules is presented. The present work is an investigation of the in vitro release kinetics of diclofenac sodium (DFS) from chitosan beads and microgranules. The in vitro release profiles of DFS from chitosan beads and microgranules are monitored using Shimadzu 1601 UV-VIS spectrophotometer. Drug release behavior of beads and microgranules has been compared. The release rate of DFS from the beads has been found to be slower in comparison to the microgranules. It may also be noted that the percent and amount of the drug release were much higher in acidic solution than in basic solution, probably due to the swelling properties of the matrix at acidic pH. PMID- 10817264 TI - Diverse mechanisms of osteoblast spreading on hydroxyapatite and titanium. AB - Hydroxyapatite (HA) is an osteoconductive implant material. We previously demonstrated that RGD peptides regulate the spreading of HOS cells on HA but not on titanium, speculating that the osteoconductivity of HA might be attributed to this RGD domain-dependent spreading of osteoblasts. To confirm this hypothesis, the molecules which regulate the spreading of HOS cells on HA and on titanium were investigated. The 50% effective dose (ED50) of RGD peptide for the spreading on HA was five fold lower comparing to titanium. Anti-alphaV integrin antibody, vitronectin, and fibronectin inhibited the spreading on HA but not on titanium. In Western blot analysis, vitronectin and fibronectin were found in components adsorbed to HA but not to titanium. Taken together, the spreading of HOS cells on HA but not on titanium requires the interaction of alphaV integrin and its ligands. The ED50 of the RGD peptides on titanium but not on HA was remarkably reduced by neuraminidase treatment, that by itself could not inhibit the spreading on both materials. This phenomenon suggests that RGD domain and sialic acid cooperatively but not independently mediate the spreading of HOS cells on titanium. Collectively, the molecules regulating the spreading on HA are apparently different from those on titanium. The spreading of osteoblasts mediated by RGD domain of vitronectin and fibronectin might contribute to the osteoconductive ability of HA. PMID- 10817265 TI - Thin film of low-crystalline calcium phosphate apatite formed at low temperature. AB - Surface modification of biomaterials to improve biocompatibility without changing their bulk properties is desired for many clinical applications and has become an emerging technology in biomaterial research and industry. In the present study, a simple method of coating the solid surfaces of metals, organic tissue matrices, glasses, inorganic ceramics as well as organic polymers with a thin film of low crystalline apatite crystals (LCA) was developed. Acidic solution containing calcium and phosphate ions was neutralized with alkaline solution to form calcium phosphate precipitates at low temperature. Precipitates of solid calcium phosphate particles were, then, removed by filtration. Concentration of free ions in the filtered ion solution which were not involved in the formation of calcium phosphate precipitate was high enough to induce the heterogeneous nucleation on the solid surfaces at low temperature. Thin layers of calcium phosphate crystals were formed on the surfaces of metals, glasses, inorganic ceramics, organic polymers including hydrophobic ones, and biological tissue matrices with this solution. The thin layer of crystals consisted of poorly crystalline calcium phosphate apatite crystals which contain high amount of labile ions like bone crystals and did not dissolve in the physiologic solutions. Various cells attached to this crystal layer and proliferated well. PMID- 10817266 TI - Bioresorbable microspheres by spinning disk atomization as injectable cell carrier: from preparation to in vitro evaluation. AB - Vesico-ureteral reflux, a common pathology in children, can be treated cystoscopically by injection of a bulking material underneath the most distal, intramural ureter, which forces the latter to do a detour, increasing its submucosal path. This increase of the length of the submucosal path of the ureter within the bladder is directly responsible for the anti-reflux effect. So far Teflon and collagen paste have been commonly used as bulking materials. We suggest replacing these materials by living tissue consisting of bladder smooth muscle, normally present at this location. The aim of this work is to provide a long-term effective treatment by producing bioresorbable microspheres which can act as a support matrix and an entrapment substance for bladder smooth muscle cells, with the goal of an in vivo transfer of the in vitro cultured cells with a minimal surgical procedure. By the use of Spinning Disk Atomization, which has specifically been developed for this purpose, we have shown two methods for the preparation of porous poly(lactic acid) microspheres with tunable sizes from 160 to 320 microm. The controlled solvent burst method has shown the advantage over the crystal leaching method in the direct creation of microspheres with large closed pores, by atomizing the polymer solution in controlled temperature conditions. Microspheres with various closed pore structures have thus been prepared. The innovation of this work is in the direct and rapid formation of porous microspheres with a pore morphology which is designed to create cavities suitable for adherence and growth of cells by adapting the temperature conditions of atomization. Injection tests have shown promising results in using these cell loaded microspheres for future non-invasive tissue engineering. PMID- 10817267 TI - Segmental bone repair by tissue-engineered periosteal cell transplants with bioresorbable fleece and fibrin scaffolds in rabbits. AB - The biological bone healing depends on the presence of osteochondral progenitors and their ability for proliferation. Isolated periosteal cells were seeded into biodegradable PGLA polymer fleece or fibrin beads and cultivated for 14 days after prior monolayer culture. On 12 New Zealand white rabbits 8 mm metadiaphyseal ulna defects were created bilaterally and subsequently filled with cell-fibrin beads, with polymers seeded with cells compared to controls with fibrin beads and polymers alone and untreated defects. A semiquantitative grading score was applied for histomorphological and radiological analysis after 28 days. Histologically intense bone formation was observed in both experimental groups with cell transplants only. The histological and radiological scoring was superior for both experimental groups. Control groups revealed only poor healing indices and untreated defects did not heal. The highest histological score was noted in the group with polymer fleeces containing periosteal cells. Applying the radiographic score system we determined a significant difference between experimental groups and controls without cells. The radiographic and histological scores for both experimental groups containing periosteal cells differed not significantly. The results strongly encourage the approach of the transplantation of pluripotent mesenchymal cells within a suitable carrier structure for the reconstruction of critical size bone defects. PMID- 10817268 TI - Treatment of rat pancreatic islets with reactive PEG. AB - Covalent attachment of polymers to cells and tissues could be used to solve a variety of problems associated with cellular therapies. Insulin-dependent diabetes mellitus is a disease resulting from the autoimmune destruction of the beta cells of the islets of Langerhans in the pancreas. Transplantation of islets into diabetic patients would be an attractive form of treatment, provided that the islets could be protected from the host's immune system in order to prevent graft rejection. If reaction of polyethylene glycol (PEG) segments with the islet surface did not damage function, the immunogenicity and cell binding characteristics of the islet could be altered. To determine if this process damages islets, rat islets have been isolated and treated with protein-reactive PEG-isocyanate (MW 5000) under mild reaction conditions. An assessment of cell viability using a colorimetric mitochondrial activity assay showed that treatment of the islets with PEG-isocyanate did not reduce cell viability. Insulin release in response to secretagogue challenge was used to evaluate islet function after treatment with the polymer. The insulin response of the PEG-treated islets was not significantly different than untreated islets in a static incubation secretagogue challenge. In addition, PEG-isocyanate-treated islets responded in the same manner as untreated islets in a glucose perifusion assay. Finally, the presence of PEG on the surface of the islets after treatment with the amine reactive N-hydroxysuccinimide-PEG-biotin (not PEG-isocyanate) was confirmed by indirect fluorescence staining. These results demonstrate the feasibility of treating pancreatic islets with reactive polymeric segments and provide the foundation for further investigation of this novel means of potential immunoisolation. PMID- 10817269 TI - Direct and indirect effects of P2O5 glass reinforced-hydroxyapatite composites on the growth and function of osteoblast-like cells. AB - Human osteoblast-like cells were plated on hydroxyapatite and P2O5-glass reinforced hydroxyapatite composite discs. They were also cultured in the presence of media obtained by incubating the discs in the absence of cells. The effects of these direct and indirect interactions were examined by measuring cell proliferation and the expression of certain key extracellular matrix antigens. One composite was found to initially delay cell growth, while the extract of a different composite appeared to down-regulate DNA synthesis. Flow cytometry analysis showed that growth directly on the discs had little effect on collagen type I, but reduced fibronectin and osteocalcin levels. The extracts of the materials generally had less effect, although one extract obtained from the glass reinforced hydroxyapatite significantly down-regulated fibronectin. These in vitro studies thus suggest that there were only few differences overall in the growth of the cells directly on the glass-reinforced compared with the hydroxyapatite discs and also only relatively small effects of the extracts on the cells. However, the flow cytometry results suggest that both the materials and the extracts may have a potentially important influence on connective tissue production, and that these effects are both material- and antigen-specific. PMID- 10817270 TI - Molecular mechanisms of improved adhesion and growth of an endothelial cell line cultured on polystyrene implanted with fluorine ions. AB - Endothelial cells derived from the bovine pulmonary artery (line CPAE, CCL 209, American Tissue Culture Collection, Rockville, MD, USA) were cultured on pristine or fluorine ion-irradiated polystyrene (5 x 10(12) or 5 x 10(14) F ions/cm2, 150 keV). At 24-h post-seeding interval, the number of cells which adhered to the ion modified polystyrene was significantly higher than on the unmodified material (+20 and +58% in cultures with the polystyrene irradiated by lower and higher ion doses, respectively). On day 7, the populations cultured on the irradiated substrates grew to higher densities, exceeding the controls at the lower and higher ion doses by 69 and 180%, respectively. The cells on ion-implanted samples were also larger (+70-95% and +90-99% at the lower and higher ion doses, respectively) and contained more protein (+16% at both ion doses). As was shown by ELISA, the polystyrene irradiated by the higher ion dose enhanced the expression of a cytoskeletal protein, vimentin (+65%) and protein of focal adhesion plaques, talin (+15%). The content of integrin alpha5beta1 (VLA-5), receptor for fibronectin, was increased at both lower and higher ion doses (+22 and +57%). In contrast to this, the content of ICAM-1 and vinculin was similar in cells grown on both pristine and ion-irradiated growth substrates. Moreover, the expression of VCAM-1 and ELAM-1 was lower by 11-14% in both ion dose groups. The present study has shown that ion implantation of polymers improves the adhesion and growth of endothelial cells without elevating the expression of immunoglobulin and selectin types of adhesion molecules. This surface modification should promote colonization of an artificial vascular prosthesis by endothelial cells and make it less vulnerable by immune system cells of the recipient. PMID- 10817272 TI - Subspecialty clinics require evidence of efficacy. PMID- 10817271 TI - Photopolymers in orthopedics: characterization of novel crosslinked polyanhydrides. AB - Novel, high modulus, degradable polymers were prepared from methacrylated anhydride monomers of tricarballylic acid (MTCA) and pyromellitylimidoalanine (MPMA-ala). Kinetic studies indicate that the time scale of photopolymerization of MTCA (< 30 s) is suitable for in vivo applications. Additionally, the tensile modulus of copolymers of these novel monomers with methacrylic anhydride (MA) ranged from 0.8 to 2.1 GPa, which lies between the modulus of trabecular and cortical bone. Degradation studies indicate that the copolymers of MTCA and MPMA ala with MA are initially surface degrading, which is important to maintaining polymer strength through the degradation process. Monomers such as these that can be rapidly polymerized using ultraviolet or blue light into high modulus degradable materials have great potential in orthopedics. PMID- 10817273 TI - Should performance of diagnostic procedures in pediatric gastroenterology be limited to pediatric gastroenterologists? PMID- 10817274 TI - Sedation for pediatric endoscopic procedures. PMID- 10817275 TI - Treatment of acute diarrhea in the new millennium. PMID- 10817276 TI - Oral rehydration therapy: a critical component in integrated management of childhood illness. PMID- 10817277 TI - Transmission of hepatitis C virus from infected mother to offspring during subsequent pregnancies. AB - BACKGROUND: Several studies have demonstrated that hepatitis C virus (HCV) may be transmitted from mother to offspring. To date, however, little is known about the risk of vertical transmission during subsequent pregnancies. The purpose of this study was to evaluate the risk of vertical HCV transmission in offspring in subsequent pregnancies of HCV infected women. METHODS: In a multicenter study, two groups of index cases were selected. Group 1 included 75 children investigated for HCV infection during prospective studies of vertical transmission. Group 2 included children born to HCV-infected mothers and found to be HCV infected, independent of studies on vertical transmission. All children in the index cases had one or more siblings. Anti-HCV, HCV-RNA (determined by polymerase chain reaction), and HCV genotype were evaluated in all the infected children, their mothers, and siblings. RESULTS: The results indicate that a mother who has already delivered an HCV-infected baby is not at greater risk of infecting her second child. Duration of maternal infection does not seem to be a risk factor in offspring infection, because HCV infection is equally distributed among first-born infants and infants of subsequent births. Because clustering of HCV infection among siblings appeared to be rare in this study, data also indirectly confirm that the risk of horizontal transmission of HCV among siblings is low. CONCLUSION: For practical purposes, the current observations indicate that mothers who have already delivered an HCV-infected child can be advised that this event does not increase the probability of infecting the second child. PMID- 10817278 TI - Contribution of villous atrophy to reduced intestinal maltase in infants with malnutrition. AB - BACKGROUND: It has been known for many years that small intestinal maltase activities are reduced in malnourished infants and in other patients with villous atrophy. The recent availability of human maltase-glucoamylase cDNA provides the opportunity to test the hypothesis that villous atrophy accounts for the reduced maltase enzyme activity in malnourished infants. METHODS: Mucosal biopsy specimens obtained for clinical evaluation of malnourished infants with poor responses to refeeding were examined by quantitative methods for enzyme activity and mRNA levels. RESULTS: Maltase activity and maltase-glucoamylase mRNA were reduced (approximately 45% of normal). When maltase-glucoamylase message was normalized to villin message, a structural protein expressed only in enterocytes, a preservation of maltase messages in surviving enterocytes was documented. The luminal glucose transporter-villin message was also preserved. CONCLUSIONS: The loss of maltase-glucoamylase message paralleled the reduction in villin message and degree of villous atrophy. The reduced maltase-glucoamylase message also paralleled sucrase-isomaltase message, previously found to be decreased in proportion to villous atrophy of malnourished infants. The data directly demonstrate, for the first time, that the terminal steps of starch 1-4 starch digestion and sucrase-isomaltase 1-6 starch digestion are decreased in malnourished infants, secondary to villous atrophy. These data in prior and present reports suggest that mechanisms underlying the chronic villous atrophy of malnutrition should be a priority for investigations in malnourished infants with slower than expected weight gain during refeeding. PMID- 10817279 TI - Objective ranking of fibrosis in standard histologic sections of human neonatal liver: applicability to alpha1-antitrypsin deficiency. AB - BACKGROUND: The etiologic heterogeneity of fibrotic liver disease has resulted in the formulation of diverse, often disease-specific, classification systems for biopsy assessment, based on tissue morphology and staining. Their qualitative nature and observer dependency remain a concern, and no classification exists for several significant conditions--for example, alpha1-antitrypsin deficiency (alpha1-ATD). The authors propose a disease- and morphology-independent numeric ranking system to objectively quantify fibrosis in a standard histologic section, based on its content of protein amino acids. This PNC system is applied to two cases of alpha1-ATD liver fibrosis. METHODS: High-performance liquid chromatography separation of the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC)-labeled acid hydrolysate of an individual needle biopsy section, followed by the calculation of specific amino acid ratios to eliminate confounding variables. RESULTS: As required by the PNC system, three numeric values were identified per tissue section, one increasing (P quotient), one decreasing (N quotient), one constant (C quotient) as fibrosis progresses, assessed by calibration against Knodell-staged samples. Generated for the alpha1-ATD sections, these three coordinates numerically referenced the degree of fibrosis in a manner that in each case was consistent with the histologic evaluation, the laboratory values, and the clinical course. CONCLUSIONS: Numeric, objective referencing of the degree of fibrosis in routine liver biopsy sections, based on the PNC system, is technically possible. PMID- 10817280 TI - Evaluation of the use of botulinum toxin in children with achalasia. AB - BACKGROUND: Achalasia is rare in children. Recently, injection of botulinum toxin into the lower esophageal sphincter has been studied as an alternative to esophageal pneumatic dilatation or surgical myotomy as treatment for achalasia. In the current study, the effects of botulinum toxin were investigated in the largest known series of children with achalasia. METHODS: Treatment for achalasia was assessed in 23 pediatric patients who received botulinum toxin from June 1995 through November 1998. Those who continued to receive botulinum toxin and did not subsequently undergo pneumatic dilatation or surgery were considered repeat responders. Results were compared with those of published studies evaluating the use of botulinum toxin in adults with achalasia. RESULTS: Nineteen patients initially responded to botulinum toxin. Mean duration of effect was 4.2 months +/ 4.0 (SD). At the end of the study period, three were repeat responders, three experienced dysphagia but did not receive pneumatic dilatation or surgery, three underwent pneumatic dilatation, eight underwent surgery, three underwent pneumatic dilatation with subsequent surgery, and three awaited surgery. Meta analysis shows that, in the current study group, the data point expressing time of follow-up evaluation versus percentage of patients needing one injection session without additional procedures (botulinum toxin injection, pneumatic dilatation, or surgery) falls within the curve for those in studies on adult patients receiving botulinum toxin for achalasia. CONCLUSIONS: Botulinum toxin effectively initiates the resolution of symptoms associated with achalasia in children. However, one half of patients are expected to need an additional procedure approximately 7 months after one injection session. The authors recommend that botulinum toxin be used only for children with achalasia who are poor candidates for either pneumatic dilatation or surgery. PMID- 10817281 TI - Milk-borne insulin with trypsin inhibitor in milk induces pancreatic amylase development at the onset of weaning in rats. AB - BACKGROUND: The physiologic significance of milk-borne hormones and growth factors for internal organs of suckling animals is poorly understood. In this study the significance of milk-borne insulin was evaluated, as well as its combination with trypsin inhibitor, and its role in the development of pancreatic digestive capacity at the time of weaning was investigated. METHODS: Experiments were performed using insulin-deficient milk formula (standard formula), insulin (20 ng/ml) formula, or insulin with trypsin inhibitor (1 U/ml) formula by a rat artificial-rearing technique. RESULTS: In 17-day-old rats administered standard formula, the plasma insulin level was as low as that in 14-day-old rats. When insulin-trypsin inhibitor formula was administered to rat pups, the plasma insulin level was significantly higher than those in rats given standard or insulin formula. In rats artificially reared on standard formula, the usual developmental increases in pancreatic amylase activity and plasma insulin concentration at the beginning of weaning did not occur. Insulin formula elevated the pups' plasma insulin concentration and amylase activity at the onset of weaning but not to the levels observed in mother-reared rats. In rats reared on insulin-trypsin inhibitor formula, the developmental increases in the plasma insulin concentration and amylase activity observed in mother-reared rats were induced. CONCLUSIONS: The present study demonstrates the necessity of milk-borne insulin for the development of pancreatic amylase during the weaning period. PMID- 10817282 TI - Management of acute gastroenteritis in Europe and the impact of the new recommendations: a multicenter study. The Working Group on acute Diarrhoea of the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition. AB - BACKGROUND: The European Society for Paediatric Gasteroenterology, Hepatology and Nutrition (ESPGHAN) issued two sets of recommendations for the treatment of infants with acute gastroenteritis (1992, 1997). The purpose of this multicentre study performed in 29 European countries was to determine how closely current treatment compares with the ESPGHAN recommendations. METHODS: The outline of the study was based on a questionnaire that addressed the management of a 6-month-old infant with acute gastroenteritis complicated by mild to moderate dehydration. National coordinators circulated the questionnaire to randomly selected primary care physicians and to hospital-based paediatricians. RESULTS: A total of 2997 questionnaires were returned, of which 1768 were from Western Europe (WE) and 1229 from Central and Eastern Europe (CEE). Eighty-four percent of responding physicians said they would follow the ESPGHAN recommendation to use oral rehydration solution (ORS) for rehydration, with 66% using an ORS containing 60 mmol/l sodium ORS. Only 16% (WE 15%, CEE 19%) would follow the guidelines and use rapid oral rehydration over 3 to 4 hours. Forty-five percent would rehydrate infants in a 3- to 6-hour period (WE 35%, CEE 60%), and 17% (WE 23%, CEE 9%) would extend the rehydration period to 12 to 24 hours. ESPGHAN recommendation of rapid reintroduction of normal feeding after 3 to 4 hours of oral rehydration would be followed by only 21% of responding physicians, and only 43% (WE 46%, CEE 38%) would start feeding with full-strength formula. However, the guideline about continuation of breast-feeding is widely followed (total 77%; WE 78%, CEE 75%). Thirty-six percent (WE 45%, CEE 23%) use a lactose-containing formula after successful oral rehydration. Contrary to the ESPGHAN guideline 35% (WE 30%, CEE 42%) would use a lactose-free formula and 19% (WE 12%, CEE 28%) a lactose and cow's milk protein-free formula. Only 37% (WE 30%, CEE 46%) of responding physicians would follow the recommendation to use ORS to replace ongoing losses from watery diarrhoea. CONCLUSIONS: The results of the survey suggest that with the exception of recommending ORS for rehydration and continuation of breast feeding during diarrhoea, a minority of responding European physicians follow the ESPGHAN guidelines for optimal management of children with acute gastroenteritis. PMID- 10817283 TI - Effects of liver transplantation on long-chain polyunsaturated fatty acid status in infants with biliary atresia. AB - BACKGROUND: The long-chain polyunsaturated fatty acid (LC-PUFA) status of infants with untreated biliary atresia (BA) is known to be poor and is correlated to the severity of the liver disease. Liver transplantation (LT) markedly increases survival of patients with BA but the extent to which this reverses poor LC-PUFA status is not known. METHODS: To explore this question, the erythrocyte (red blood cell, RBC) phospholipid content of eight infants with BA who underwent LT was determined 2 months after an initial portoenterostomy, immediately before LT, and 6 and 12 months after LT. Before LT, all infants were fed a protein hydrolysate formula containing medium-chain triglycerides and essential fatty acids. Afterward, they were fed a normal diet for age. The RBC phospholipid content at each time point was compared with that of 28 age-matched control infants. RESULTS: Just before LT, median RBC phospholipid content of C20:4n-6, C20:5n-3, and C22:6n-3 was 25%, 48%, and 30% lower, respectively, than that observed in age-matched control infants. After LT, the RBC phospholipid content of most fatty acids reached normal values by 6 months. However, that of C20:4n-6 and C22:6n-3 contents remained 5% and 15% lower, respectively, than in normal control infants. Twelve months after LT, C20:4n-6 content remained lower than in normal children, but that of C22:6n-3 did not differ. The ratio of C20:3n 6/C20:4n-6, a reflection of delta-5 desaturase activity, was abnormal compared with normal children before LT (0.17 vs. 0.10, P < 0.009) but normalized by 6 months after LT (0.11 vs. 0.10, not significant). CONCLUSIONS: These data show that the abnormal LC-PUFA status of children with BA improves after LT but is not entirely reversed within a year after surgery. They suggest that the abnormal status before LT may be secondary, in part, to low delta-5 desaturase activity. The extent to which a different pre- and/or post-LT diet can prevent PUFA deficiency and/or hasten recovery of PUFA status remains to be determined. PMID- 10817284 TI - Early development of human gastric H,K-adenosine triphosphatase. AB - BACKGROUND: Little is known about the early development of the gastric acid secretion in human neonates. The purpose of this study was to examine the early development of gastric H,K-adenosine triphosphatase (ATPase) by analyzing human gastric biopsy specimens. METHODS: Eighty-eight neonates from week 25 to week 42 of gestation who were treated in a neonatal intensive care unit underwent gastroscopy with biopsy specimens obtained from the corpus. The expression of gastric H,K-ATPase protein in the gastric biopsy specimens was assessed by Western blot analysis, using an antibody directed against the gastric H,K-ATPase. The amount of H,K-ATPase expressed was compared with age, gender, clinical factors, diseases, and the macroscopic and histologic findings at endoscopy. RESULTS: The expression of human gastric H,K-ATPase increased significantly with gestational age. There was a significant increase in the expression of gastric H,K-ATPase during the first 82 days after birth. Boys had a significantly higher expression of gastric H,K-ATPase than girls did, when it was adjusted for gestational and postnatal age. Neither the clinical features nor treatments showed significant correlations with the expression of human gastric H,K-ATPase when controlling for gestational and postnatal age. CONCLUSIONS: This study shows that human gastric H,K-ATPase is expressed from week 25 of gestation, which agrees with earlier findings of gastric pH in preterm infants. The amount of enzyme expressed increases with gestational and postnatal age. The authors speculate that the susceptibility to gastric lesions seen in neonates is not related to the amount of H,K-ATPase. However, studies elucidating the ontogeny of gastric mucosal defense mechanisms are warranted. PMID- 10817285 TI - Effect of intravenous lipid emulsions on hepatic cholesterol metabolism. AB - BACKGROUND: Total parenteral nutrition offers the chance of survival to children who have had extensive gut resections or gut failure. However, in infants it is often associated with serious complications including cholestatic liver disease. The causes of these complications remain unclear, although it has been suggested that the lipid emulsions used in total parenteral nutrition may be responsible. METHODS: An in vitro system was developed to study the effect of lipid emulsions on hepatic cholesterol metabolism using cultured hepatocytes. RESULTS: Incubations of Hep G2 cells with medium containing Intralipid (Pharmacia and Upjohn, Milton Keynes, UK) demonstrated that the fat emulsion mediated a powerful dose-dependent but reversible inhibition of cholesterol uptake. In addition Intralipid was shown to stimulate the efflux of cholesterol from Hep G2 cells. The component or components of the Intralipid responsible for these effects and the mechanism by which they act remain to be established. CONCLUSIONS: Intravenous lipid emulsions may interfere with hepatic cholesterol metabolism in vivo. This may have implications for the development of total parenteral nutrition-associated cholestasis in neonates. PMID- 10817287 TI - Normal growth velocity before diagnosis of celiac disease. AB - BACKGROUND: Clinical experience leads us to believe there may be patients with celiac disease who have not yet been treated, with a normal physical growth. METHODS: To evaluate height velocity of patients with confirmed celiac disease before their diagnosis and treatment, anthropometric measurements taken by the general pediatricians in charge of the primary care of the patients before they were sent to the authors' hospital were studied. Forty-two growth periods (available velocities) were measured at varying intervals (ranging from 6 to 27 months) in 23 patients aged 0.1 to 10.66 years were analyzed. RESULTS: All patients studied during the first semester of life (n = 5) showed normal growth velocity, and 6 of 10 patients showed normal growth velocity during the second semester of life. Ten of 12 patients between the ages of 1.0 and 1.99 years of age showed normal height velocity, and 7 of 9 patients aged 2.0 to 10.66 years showed normal height velocity. Normal height velocities were found not only during the first year of life, but also in children aged 1 to 8 years. CONCLUSIONS: Results should alert pediatricians and those in gastroenterology and growth clinics. In the latter case, norms for studying children who have short stature but are growing at a normal rate should not be a condition for excluding a child from screening for celiac disease. PMID- 10817286 TI - Effect of type of early infant feeding on fatty acid composition of plasma lipid classes in full-term infants during the second 6 months of life. AB - BACKGROUND: Previously, the authors found significantly higher arachidonic and docosahexaenoic acid values in plasma lipids in 2-month-old full-term infants fed human milk than in those receiving formula. This is the report of data obtained in full-term infants during the second half of the first year of life. METHODS: Healthy, full-term infants fed human milk (n = 12) or formula without preformed long-chain polyunsaturated fatty acids (n = 12) were investigated. Fatty acid composition of plasma lipid classes was determined by high-resolution capillary gas-liquid chromatography. RESULTS: Linoleic acid acid values in plasma phospholipids (18.5 [3.94] vs. 20.79 [4.34]) and gamma-linolenic acid values in plasma cholesteryl esters (0.17 [0.09] vs. 0.27 [0.20]) and triacylglycerols (0.27 [0.18] vs. 0.46 [0.27]) were significantly (P < 0.05) lower in breast-fed infants than in those receiving formula. Data are percentage weight by weight shown as median (range from 1st to 3rd quartile) for breast-fed vs. formula fed infants, respectively. In contrast, arachidonic acid values in plasma phospholipids (10.05 [2.90] vs. 7.03 [1.87]; P < 0.01), cholesteryl esters (7.54 [3.58] vs. 4.09 [1.81]; P < 0.05), and triacylglycerols (1.28 [0.84] vs. 0.80 [0.39]; P < 0.05), as well as docosahexaenoic acid values in plasma phospholipids (1.92 [0.36] vs. 1.02 [0.31]; P < 0.001), cholesteryl esters (0.39 [0.13] vs. 0.15 [0.13]; P < 0.001), and triacylglycerols (0.17 [0.17] vs. 0.09 [0.04]; P < 0.01) were significantly higher in infants fed human milk than in those receiving formula. CONCLUSION: Healthy, full-term infants fed formula without preformed dietary long-chain polyunsaturated fatty acids are unable to match the arachidonic and docosahexaenoic acid status of breast-fed infants even during the second half of the first year of life. PMID- 10817288 TI - Clinical quiz. Ascaris infection. PMID- 10817289 TI - Postsurgical recurrences in Crohn's disease: why, when and how to prevent them. PMID- 10817290 TI - New insight into left ventricular function in tricuspid atresia after total cavopulmonary connection: a three-dimensional echocardiographic study. AB - BACKGROUND: In patients with tricuspid atresia palliated by construction of a total cavopulmonary connection, both pulmonary and systemic circulations depend on the performance of the dominant left ventricle. When estimating the volume of such ventricles using cross-sectional echocardiography, it is necessary to make assumptions concerning the geometry of the ventricular shape. This is avoided by three-dimensional echocardiography, which provides direct volumetric data. Our purpose was to apply this new method to quantify left ventricular volumes and function in patients with tricuspid atresia after construction of a total cavopulmonary connection. METHODS: We studied ten patients (median age: 8 years) with tricuspid atresia who had undergone a total cavopulmonary connection, comparing them with 10 normal children matched for age, sex and size. The three dimensional echocardiography was performed with electrocardiographic and respiratory gating. A new transthoracic integrated probe designed for small children was used with a rotational scanning increment of 3 degrees. The 60 slices obtained from the ventricular cavity were stored and formatted in a commercial system (TomTec). End-diastolic and end-systolic volumes, stroke volume and ejection fraction were calculated after manual tracing of the endocardial surfaces. The volumes were indexed to the body surface area. RESULTS: As seen in the reconstructions, the dominant left ventricle in tricuspid atresia had a spherical shape, whereas the normal left ventricle is oblong. The left ventricular volumes and function in tricuspid atresia were 54+/-4 ml/m2 (end diastolic volume), 28+/-3 ml/m2 (end-systolic volume), 26+/-7 ml/m2 (stroke volume) and 48+/-6% (ejection fraction). These volumes were not different from those obtained in the controls (p = NS). The left ventricular stroke volume and ejection fraction in 10 patients with tricuspid atresia were lower than those calculated for the controls (p < 0.05). CONCLUSIONS: Three-dimensional echocardiography provides a quantitative insight into the pathophysiologic function of the dominant left ventricle in tricuspid atresia after construction of a total cavopulmonary connection. PMID- 10817291 TI - Spectrum and influence of hypoplasia of the left heart in neonatal aortic coarctation. AB - Obstruction of the left ventricular outflow tract may be associated with hypoplasia of the left heart, which importantly influences the options for treatment. Although the influence of the size of the left heart on the outcome for critical aortic stenosis has been described, less is known about the spectrum of such hypoplasia seen with neonatal aortic coarctation, and how this influences outcome. To determine, first, the spectrum and influence of hypoplasia of the left heart in neonatal coarctation, second, if the previously described critical values for adequacy of the left heart in neonates with critical aortic stenosis are applicable to neonates with coarctation, and, third, if any of the variables or associated abnormalities are risk factors for recoarctation, we studied 63 neonates who underwent repair of coarctation. From the initial echocardiogram, we measured multiple structures in the left heart, and calculated a score for adequacy as has been done for critical aortic stenosis. The sizes were compared to previously reported minimal values. We then analyzed the influence of the variables and the associated anomalies on outcome. There were no deaths. There was a broad spectrum of sizes that did not correlate with the need for re intervention. The calculated score for adequacy would have predicted survival in only 56% of the patients, and 73% of the neonates had at least one parameter measured in the left heart below the previously reported minimal values. There is, therefore, a broad spectrum of sizes for the left heart in neonates with aortic coarctation that is not predictive of outcome. Minimal sizes, and the score for adequacy used for critical aortic stenosis, are not applicable to neonates with coarctation. PMID- 10817292 TI - Physical activity in children and adolescents with congenital heart disease. Aspects of measurements with an activity monitor. AB - The aim of the present study was to evaluate different aspects of a device designed to monitor physical activity. Measurements of different axes and placement of the monitor were tested using a treadmill with ranging increments in incline or speed. The monitor was also used to assess the level of physical activity among children and adolescents with congenital heart disease and in healthy controls at the same age. The results indicate that the monitor is a valid and reproducible instrument for measurements of physical activity. The study revealed that the level of activity was higher for healthy boys than healthy girls (p<0.0001). Boys with congenital heart disease also displayed higher values compared to girls with congenital heart disease, although the difference was not significant (p=0.067). Healthy boys revealed a significantly higher level of activity than did boys with congenital heart disease (p=0.003), but no such difference was found in girls (p=0.757). Nor were any differences found between younger and older individuals among patients with congenital heart disease. Young healthy controls, however, showed significantly higher levels of activity than their older counterparts. There were differences in activity monitored during the week, with lower activity in the weekends, but the activity on the same day in different weeks seemed stable. Neither were there any differences between measurements over whole weeks. The results indicate that the Computer Science & Application monitor is a valid instrument for assessing physical activity. The monitor may also be used, therefore, to validate the levels of physical activity level in children with congenital heart disease after medical and surgical treatment. PMID- 10817293 TI - Effect of physical training in children and adolescents with congenital heart disease. AB - In order to test the effect of systematic supervised physical training, we divided a total of 129 children and adolescents with congenital heart disease into a group undergoing intervention and a control group. All patients underwent exercise tests, measurements of physical activity, and a survey of psychosocial factors. An improvement in uptake of peak level of oxygen was observed after intervention. There was also an improvement in physical activity in both groups measured by a monitor, although this was significant only in those with intervention. The psychosocial scales measured by the Child Behavior Checklist showed a decrease in internalizing scores for those subjected to intervention. This was decreased due to decreased withdrawal and somatic complaints. In conclusion, we recommend systematic supervised training, including testing of routine follow-ups, in patients with congenital heart disease. PMID- 10817294 TI - Non-invasive determination of the systolic peak-to-peak gradient in children with aortic stenosis: validation of a mathematical model. AB - Doppler derived systolic pressure gradients have become widely applied as noninvasively obtained estimates of the severity of aortic valvar stenosis. There is little correlation, however, between the Doppler derived peak instantaneous gradient and the peak-to-peak gradient obtained at catheterisation, the latter being the most applied variable to determine severity in children. The purpose of this study was to validate a mathematical model based on data from catheterisation which estimates the peak-to-peak gradient from variables which can be obtained by noninvasive means (Doppler derived mean gradient and pulse pressure), according to the formula: peak-to-peak systolic gradient = 6.02+/ 1.49*(mean gradient)-0.44*(pulse pressure). Simultaneous cardiac catheterization and Doppler studies were performed on 10 patients with congenital aortic valvar stenosis. Correlations between the gradients measured at catheter measured, and those derived by Doppler, were performed using linear regression analysis. The mean gradients correlated well (y = 0.67 x +11.11, r = 0.87, SEE = 6 mm Hg, p = 0.001). The gradients predicted by the formula also correlated well with the peak to-peak gradients measured at catheter (y = 0.66 x +14.44, r = 0.84, SEE = 9 mm Hg, p = 0.002). The data support the application of the model, allowing noninvasive prediction of the peak-to-peak gradient across the aortic valvar stenosis. PMID- 10817295 TI - Isolated origin of the left subclavian artery from the left pulmonary artery. AB - We describe two children with isolated origin of the left subclavian artery from the left pulmonary artery detected by echocardiography during the assessment of their congenital cardiac malformations. Both patients demonstrated pre-operative evidence of subclavian steal. This entity results from persistence of the dorsal segment of the sixth left arch, with regression of the left fourth arch and interruption of the left dorsal arch distal to the origin of the seventh left intersegmental artery. The significance of this finding relates to the potential for pulmonary overcirculation, which could have significant post-operative ramifications if not detected prior to surgical repair of an associated cardiac malformation. This entity differs from cases with a right aortic arch and aberrant left subclavian artery which has the potential to form a vascular ring, unlike cases with isolated origin of the left subclavian artery from the pulmonary artery that do not cause compression of the airway. PMID- 10817296 TI - Left ventricular function in adolescents and adults with restrictive ventricular septal defect and moderate left-to-right shunting. AB - BACKGROUND: The long-term haemodynamic effects of a restrictive ventricular septal defect permitting moderate left-to-right shunting are not known. PATIENTS AND METHODS: Echocardiographic measurements of left heart dimensions and function were compared between a group of 9 older children and adults (median age 21 years, range 12-24.5 years) having restrictive ventricular septal defects (median Qp/Qs 1.7, range 1.4-2.1) and a group of 10 age matched controls. RESULTS: Left ventricular mass indexed to body surface area was significantly greater in subjects than in controls (102+/-29 vs. 75+/-13 g/m2, p=0.02), although there was no significant difference in the ratio of mass to volume. There were no significant differences between indexes of left ventricular systolic function. Ratios of peak early to late diastolic ventricular filling were lower in those with septal defects (1.5+/-0.3 vs. 1.8+/-0.3, p=0.03), but there were no differences in other indexes of diastolic function. CONCLUSIONS: Resting left ventricular function does not appear to have deteriorated by early adult life in patients with restrictive ventricular septal defects and moderate volume loading. This would support a continued conservative approach to management in these patients. PMID- 10817297 TI - Neonatal pulmonary hypertension during extracorporeal membrane oxygenation. AB - OBJECTIVES: This prospective study was designed to monitor severe pulmonary hypertension during extra corporeal membrane oxygenation using echo Doppler variables. BACKGROUND: All neonates treated with extracorporeal membrane oxygenation also have severe pulmonary hypertension. A study which monitors the reaction of the pre-existing pulmonary hypertension during extracorporeal oxygenation by frequent sampling of those variables related to pulmonary pressure is still lacking. Such a study is necessary to analyze the complex haemodynamic changes in patients undergoing extracorporeal membrane oxygenation. METHOD: In 29 neonates, we estimated pulmonary arterial pressure using peakflow velocity of regurgitation across the tricuspid- and pulmonary valve, peakflow velocity of shunting across persistent arterial ductus, and systolic time intervals of the right ventricle. Correlation between the several estimations of pulmonary arterial pressure were analysed with the Spearman correlation coefficient. RESULTS: Systolic pulmonary arterial pressure measured by the velocity of tricuspid regurgitation illustrated severe pulmonary hypertension prior to extra corporeal membrane oxygenation (mean 63 mmHg, sd 20). Similar levels for the systolic pulmonary arterial pressure could be derived (mean 73 mmHg, sd 17) from ductal shunting. A fair correlation of 0.76 (p< 0.002) could be demonstrated. Pulmonary hypertension responded well and quickly to treatment by extra corporeal membrane oxygenation, with reductions within 24 hours to mean systolic levels of 35 mmHg, sd 23. This very early reaction has not previously been demonstrated and could be of importance in defining parameters for weaning from cardiopulmonary bypass. Diastolic pulmonary arterial pressure was investigated because of its relation to vascular resistance. It proved more difficult to measure because of the low incidence of pulmonary regurgitation. Derived diastolic pressures did not show any good correlations. CONCLUSION: Pulmonary hypertension is well documented prior to extra corporeal membrane oxygenation and response very quickly to the institution of treatment. Ultra sound techniques are indicated at the bedside, and prove useful in monitoring pulmonary blood pressure during the procedure. PMID- 10817298 TI - Isomeric arrangement of the left atrial appendages and visceral heterotaxy: two atypical cases. AB - We present two cases in which there was isomeric arrangement of the left atrial appendages in association with multiple spleens and complete heart block. In both of these, the venous connections were normal. In one case the arrangement of the bronchial tree and the lungs was normal, the liver was left-sided, and the stomach was right-sided. In the other case, there was left isomeric broncho pulmonary morphology, and both liver and stomach were right-sided. These unusual combinations show the need for full description of the morphology of other organs, and the venoatrial connections, in cases with isomeric arrangement of the atrial appendages. They also demonstrate that connection of the inferior caval vein cannot be taken as a reliable marker of an atrium having a morphologically right appendage. PMID- 10817299 TI - Common arterial trunk with discordant atrioventricular connections. PMID- 10817300 TI - Anomalous origin of the main stem of the left coronary artery from the non-facing sinus of valsalva associated with sudden death in a young athlete. AB - A young female athlete is described with anomalous origin of the main stem of the left coronary artery from the non-facing sinus of Valsalva who sustained myocardial infarction and died suddenly after physical exertion. Autopsy findings illustrated the mechanistic importance of acute angle take-off of the left main coronary artery and a slit-like orifice, which was likely compressed and obstructed by acute expansion of the aortic wall. This rare type of coronary anomaly has been regarded as having little clinical significance, but it can lead to sudden cardiac death under physical exertion. PMID- 10817301 TI - Dependence on a respiratory ventilator due to an atrial septal defect. AB - An atrial septal defect is commonly thought of as a benign cardiac lesion especially in infancy. The haemodynamic consequences for the lungs, nonetheless, can be comparable to that produced by patency of the arterial duct. In a preterm boy, this lesion led to the development of broncho-pulmonary dysplasia, and dependence on a respiratory ventilator. He could be extubated shortly after surgical closure of the septal defect. PMID- 10817303 TI - Intermittent complete closure of the arterial duct. AB - Intermittent functional closure of a patent duct has been reported both clinically and echocardiographically. We describe the case of a 6-year-old with intermittent complete closure of a patent duct occurring during attempted transcatheter closure. Recurrence of clinical and echocardiographic signs prompted restudy. A patent duct of moderate size was demonstrated and successfully occluded by coil embolization. The interventional aspects of this unusual condition are discussed. PMID- 10817302 TI - Aortic vasculopathy with aneurysm: a rare cause of fetal hydrops. AB - An hydropic fetus seen at 28-weeks gestation had a saccular aortic aneurysm in the descending thoracic aorta. Histology disclosed marked fibrointimal hyperplasia, thrombus, and attenuation of the tunica media. The remainder of the descending thoracic aorta showed fibrointimal hyperplasia. We speculate that the narrowed lumen and rigid aortic wall resulting from this vasculopathy provided an increased afterload leading to cardiac failure. PMID- 10817304 TI - Fetal hydrops due to a tachyarrhythmia progressing to organic pulmonary stenosis. AB - We discovered supraventricular tachycardia with advanced hydrops in the setting of normal cardiac anatomy at 26 weeks of gestation which resolved successfully following administration of digoxin and flecainide to the mother. A female baby was born after a premature rupture of the membranes at 30.6 weeks. The neonate was in sinus rhythm, showed a progressive regression of right ventricular insufficiency, but developed signs of acquired pulmonary valvar and subvalvar stenosis at 2 months of age. PMID- 10817305 TI - Diagnosis and natural history of isolated congenital pulmonary regurgitation in fetal life. AB - We describe a rare instance of isolated pulmonary regurgitation caused by a dysplastic pulmonary valve which was detected prenatally. Fetal echocardiography demonstrated severe pulmonary regurgitation, and progressive cardiomegaly because of right ventricular volume overload. After birth, conservative therapy was successful in alleviating the pulmonary vascular resistance, and the pulmonary regurgitation gradually decreased. PMID- 10817306 TI - Influence of mechanical ventilation and inhalation of pulmonary vasodilators, upon pulmonary blood flow and pulmonary vascular resistance. PMID- 10817307 TI - Public health in transition. PMID- 10817308 TI - Inequalities in life expectancy in Lithuania by level of education. AB - The purpose of this study was to examine educational inequalities in life expectancy of the Lithuanian population. The life-tables by level of education were calculated on the basis of the individual records of the 1989 census, which were linked to the death records of males and females, aged 25-70 years. In comparison with the group with university education, the life expectancy of males with primary or lower education was 11.7 years shorter, and of females 4.3 years shorter. The greatest impact of educational differentials on life expectancy was the inequality found in the mortality of the population, aged 25-44 years. Sex differences in life expectancy were greatest among those with primary or lower education. External causes of death contributed most to educational differences in life expectancy of males, whereas cardiovascular diseases had a major impact to educational differences in females. PMID- 10817309 TI - Do early unemployment and health status among young men and women affect their chances of later employment? AB - The aim of this 5-year prospective study was to investigate the risk of future unemployment among young people who had experienced early unemployment, and to examine whether this risk was influenced by their health status. A total of 1,083 pupils in the final year of compulsory schooling were included in the cohort. The non-participation rate was 2%. At the time of a five-year follow-up, of those who were unemployed during the first two-year period, 71% of the men and 49% of the women were unemployed, had recent experience of unemployment, or were outside the labour market. The relative risk of being unemployed was 2.39 for men (95% CI 1.85-3.10) and 1.76 for women (95% CI 1.25-2.48) among those who had experienced early unemployment compared with those who had been in Youth Opportunities Programmes (YOP) or in work. Young women in YOP had the same risk of later unemployment as those who had experienced early unemployment, while young men in YOP did not have increased risk. Health status and health behaviour had only a minor influence on the risk of unemployment. PMID- 10817310 TI - Stable and dynamic differences in injury mortality between the Nordic countries. What do they say about inherent national characteristics with regard to risk? AB - The aim of this study was to analyse long-term trends and patterns in injury mortality between the Nordic countries and to distinguish differences that are relatively temporary from those that are more durable. Both total injury mortality and special subcategories were examined. Some stable similarities and differences were found. Two kinds of injuries are distinguished: those that largely have a social genesis and those to which environmental causality applies. The general pattern is that there has been an increase in injuries with a social genesis and a decrease in those with an environment-related genesis. Finland has high rates of both types of injuries. Denmark has high rates of injuries with a social genesis, but low rates of those with an environmental genesis. By contrast, Norway has high rates of injuries with an environmental genesis and low rates of those with a social genesis. Sweden has low or medium rates of both types of injuries. PMID- 10817311 TI - Family caregiving in dementia--an analysis of the caregiver's burden and the "breaking-point" when home care becomes inadequate. AB - The burden of caregivers of patients suffering from of Alzheimer type dementia (DAT) and vascular dementia (VD) was analysed at the critical time, the "breaking point", when home care becomes insufficient and/or inadequate and the caregiver burden has probably reached its upper limit. Primary family caregivers of 39 DAT and 40 VD patients who were being considered for relocation into group-living units were studied. Total caregiving burden and different aspects of the burden: general strain, isolation, disappointment, and emotional involvement, were correlated with the patients' diagnoses, abilities, and symptoms. Closer kinship to the patient imposed a heavier burden. The caregiver's gender, social class, and previous institutionalization of the patient did not influence the caregiver burden. There was no significant correlation between the patients' ADL ability or cognition and the burden. A higher level of disappointment was found among the VD carers. Different symptomatology in patients of the two diagnostic groups was related to special aspects of the burden. Multiple regression analysis showed that the amount of caregiving time each week and impaired sense of own identity, misidentifications, clinical fluctuations, and nocturnal deterioration in the patients predicted the breaking-point. PMID- 10817312 TI - Association of Helicobacter pylori infection with lifestyle, chronic disease, body-indices, and age at menarche in Danish adults. AB - This study examines the association between lifestyle factors, chronic disease, body-indices, and the seroprevalence of Helicobacter pylori infection in Danish adults. The relationship between age at menarche and H. pylori infection is also assessed. A random sample of 3,608 Danish adults completed a questionnaire about lifestyle factors (smoking habits, alcohol consumption, and coffee and tea intake), doctor diagnosed chronic diseases (heart conditions, diabetes, chronic bronchitis, and hypertension), menarche, and socio-demographic factors. A total of 2,913 participants were eligible for the present study. Electrocardiographs were recorded and blood pressure and serum lipid levels (HDL cholesterol, triglyceride, and cholesterol) were measured. Height and weight were determined and body mass index (BMI) calculated. Sera were analysed with an enzyme-linked immunosorbent assay for the presence of H. pylori specific IgG antibodies. The seroprevalence of H. pylori infection was associated with weekly alcohol intake > or = 6 drinks (odds ratio 0.7, 95% confidence interval 0.6-0.9) due to a low rate of H. pylori infection among wine drinkers (odds ratio 0.6, 95% confidence interval 0.5-0.7). No associations were found with smoking habits or serum lipids. People with upper quartile BMI (> or = 26.8 kg/m2) were more likely to be seropositive for antibodies to H. pylori (odds ratio 1.6, 95% confidence interval 1.1-2.4). Chronic bronchitis (odds ratio 1.6, 95% confidence interval 1.1-2.5) and unspecified heart condition (odds ratio 2.0, 95% confidence interval 1.1-3.3) was more often seen in IgG seropositive women than in uninfected women. The likelihood of being seropositive for IgG antibodies to H. pylori increased with age at menarche (odds ratio per year 1.10, 95% confidence interval 1.02-1.19). Previously reported associations with age and socioeconomic status were confirmed. We conclude that wine drinking is associated with lower rates of H. pylori infection in Danish adults. The seroprevalence of H. pylori infection is increased in people with high BMI. H. pylori infection may relate to a history of late menarche and chronic bronchitis in Danish women. PMID- 10817313 TI - Cost-effectiveness of an innovative four-year post-discharge programme for elderly patients--prospective follow-up of hospital and nursing home use in project elderly and randomized controls. AB - The cost-effectiveness of a post-discharge programme on the use of hospital care and the continuity of care was assessed in an elderly cohort (n = 204) discharged from the city hospital. The participation rate was 97.6%, and the patients were aged 75 years or over and lived alone. The randomized controls (n = 204) received standard aftercare. During the follow-up the costs of university hospital care decreased by 52% in the intervention group and by 24% in the control group per patient year, compared with the costs in the year preceding the project. This happened despite the higher morbidity in the intervention group in terms of fractures and the use of university hospital care in the year preceding the project. There was also a tendency in the intervention group for the previous non users of university hospital care to remain non-users during the follow-up. The costs of city hospital care increased by 16% and 5%, and of all hospital care by 1.3% and 0.2%, respectively. There were no differences in admissions to permanent care in the nursing homes. The intervention group did not make their first contact with the hospitals or permanent care in nursing homes earlier than the control group during the follow-up. The co-operation between hospital and domiciliary care and voluntary workers was well-suited to the innovative care of the elderly people. PMID- 10817314 TI - The organizational pendulum--healthcare management in Sweden 1865-1998. AB - In recent years, there have been a number of large-scale changes in the organization of healthcare in Sweden, as in many other countries. From a longer time perspective, however, there has been a more or less permanent oscillation between centralization and decentralization, and also between competition and co operation, in Swedish healthcare organizations. In fact, there seems to be something like a pendulum moving all the time between opposite forms of organization. The frequency of these changes has been increasing during the last fifteen years, as different organizational models have replaced each other at shorter and shorter intervals. This development is partly due to the increasing rate of change in modern society, but partly also due to an element of fashion in the area of management. There is, however, a growing change fatigue in many healthcare organizations, and also increasing demands for empirical evidence on the efficiency and effectiveness of different organizational models before they are introduced into the healthcare system. PMID- 10817315 TI - Suicidal ideation and suicide attempts among adult Danes. AB - This study describes the prevalence of suicidal ideation and attempted suicide in a representative survey among adults in Denmark and gives the proportion of people reporting a suicide attempt that results in contact with the health care system. The data for the 1994 Danish national health interview survey were collected by personal interview and a self-administered questionnaire. A subsample of 1362 individuals participated in the part of the survey that addressed suicidal behaviour (64% of the random sample). The results show that 6.9% reported having had suicidal thoughts within the past year. Averaging across all age groups the overall prevalence of people reporting ever having made a suicide attempt was 3.4% and the one-year prevalence was 0.5%. Suicidal ideation was more prevalent among young people than among older people, whereas ever attempted suicide showed no age gradient. Both suicidal ideation and ever attempted suicide were more prevalent among economically inactive people (e.g. unemployed) and among unmarried or divorced people. We estimate that 50-60% of suicide attempts reported in a representative, national survey become known to the healthcare system. PMID- 10817316 TI - Social support and physical and psychological recovery one year after myocardial infarction or coronary artery bypass surgery. AB - The aim of this study was to evaluate the role of different support factors supposed to explain physical and psychological recovery after myocardial infarction (MI) or coronary artery bypass surgery (CABS). The subjects comprised 147 MI patients and 159 CABS patients. Support factors included formal services, semi-formal assistance, and informal social support. The outcome measures used for analysis were functional activities level (Duke Activity Status Index, DASI), physical working capacity, anxiety, and depression one year after MI or CABS. In general, support factors had a limited role in this study. The patient's functional and psychological status at three months was the main determinant to recovery at one year. The outcome factors measured at three months explained 36 56% of their variance at one year, and the support factors increased the explanatory power by 0-10%. The support model employed in this study revealed that some single factors may have a positive or negative role in the recovery after MI or CABS. PMID- 10817317 TI - Changes in condom use after HIV diagnosis. AB - We investigated changes in condom use after HIV diagnosis. The study population comprised 78 asymptomatic HIV-infected subjects recruited from a clinic in Oslo, Norway, during 1988-92. In total, 240 follow-up visits were carried out. The response variable was repeated measurements of whether or not condoms were always used during anal/vaginal intercourse at follow-up. The explanatory variables were: time from HIV diagnosis to follow-up, exposure group (heterosexually infected, homosexual men, or infected through injecting drug use), time of HIV diagnosis (before 1987 or later), and history of condom use before HIV diagnosis. Random effects logistic regression analysis was used to study incremental changes in condom use, accommodating an unbalanced repeated measurement design. The use of condoms increased monotonously over time after HIV diagnosis in all exposure groups. Condom use was least likely among injecting drug users, subjects diagnosed before 1987 and subjects without a history of frequent condom use before HIV diagnosis. PMID- 10817318 TI - Classification of non-respondents in a population-based tobacco cessation contest -"Quit and Win". PMID- 10817319 TI - Increase in the prevalence of common mental disorders during an upswing in the national economy. PMID- 10817320 TI - Crush injuries of the pelvis. PMID- 10817321 TI - Video-assisted thoracoscopic introduction of talc in the treatment of recurrent spontaneous pneumothorax. AB - OBJECTIVE: To present our experience of using a video-assisted thoracic surgical (VATS) technique to treat spontaneous pneumothorax. DESIGN: Retrospective study. SETTING: Teaching hospital, The Netherlands. SUBJECT: 34 patients with recurrent spontaneous pneumothoraces. INTERVENTION: Combined bullectomy, parietal pleural abrasion, and spraying with talc using the Tissuecol (Immuno) spray device using a VATS technique. Measurement of pain by visual analogue scale (VAS) and analgesic requirements. MAIN OUTCOME MEASURES: Duration of drainage and hospital stay, pain, and morbidity. RESULTS: Patients were followed up after 4 weeks and 1 year. Median drainage time postoperatively was 2 days (range 1-3) and duration of hospital stay 3 days (range 2-5). One pneumothorax recurred and one wound became infected. Patients scored a median of 4 (range 1-9) on the VAS for pain at 4 weeks, and 2 (range 0-6) at 1 year. CONCLUSION: Bullectomy combined with pleural abrasion and the introduction of talc using a VATS technique is safe and effective, and requires only a short hospital stay. PMID- 10817322 TI - Truncal ligation of the inferior thyroid arteries does not affect the incidence of hypocalcaemia after thyroidectomy. AB - OBJECTIVE: To assess the effect of truncal ligation of the inferior thyroid arteries during bilateral subtotal thyroidectomy on the incidence of hypocalcaemia after thyroidectomy. DESIGN: Prospective non-randomised study. SETTING: Teaching hospital, Turkey. SUBJECTS: 216 patients who had bilateral subtotal thyroidectomy for non-toxic nodular goitre between 1990 and 1996. INTERVENTIONS: The trunk of the each inferior thyroid artery was simply ligated during bilateral subtotal thyroidectomy. MAIN OUTCOME MEASURES: Clinical examination, and measurement of serum calcium, ionised calcium, and inorganic phosphate concentrations before and after operation. RESULTS: Four patients (2%) had low concentrations of total and ionised calcium during the postoperative period. On physical examination three of them had spasms of the facial nerve, as seen in tetany. They were given calcium supplements orally and their laboratory results returned to reference ranges within 180 days. No patients developed permanent hypocalcaemia. CONCLUSION: Truncal ligation of the inferior thyroid arteries during bilateral subtotal thyroidectomy has no effect on the incidence of hypocalcaemia after thyroidectomy. PMID- 10817323 TI - Limited role of meta-iodobenzylguanidine scintigraphy in imaging phaeochromocytoma in patients with multiple endocrine neoplasia type II. AB - OBJECTIVE: To compare diagnostic applicability of combined computed tomography (CT) and magnetic resonance imaging (MRI), with that of meta-iodobenzylguanidine (MIBG) scintigraphy in the preoperative localisation of MEN II related phaeochromocytoma. DESIGN: Retrospective study SETTING: University hospital, The Netherlands. MATERIALS: 17 patients with MEN II patients (33 adrenal glands) who were operated on for phaeochromocytoma. MIBG scintigraphy, CT and MRI were used to localize phaeochromocytoma. Histopathologically, an adrenomedullary lesion more than 1 cm in size was classified as a phaeochromocytoma. MAIN OUTCOME MEASURES: Sensitivity, specificity, and diagnostic accuracy of combined CT and MRI and MIBG scintigraphy, compared with histopathological findings. RESULTS: Sensitivity of combined CT and MRI (27 adrenal glands) was 87%, with a specificity of 100% and a diagnostic accuracy of 89%. MIBG scintigraphy (31 adrenal glands) had a sensitivity of 92%, a specificity of only 17%, and a diagnostic accuracy of 77%. CONCLUSION: If unilateral adrenalectomy is done when only one adrenal gland contains a phaeochromocytoma, then MRI should be the method of choice for localising MEN type II related phaeochromocytoma. MIBG scintigraphy can be restricted to those patients in whom MRI does not show a tumour. PMID- 10817324 TI - Incidence of glove perforations in gastrointestinal surgery and the protective effect of double gloves: a prospective, randomised controlled study. AB - OBJECTIVE: To investigate the incidence of glove perforations during various types of gastrointestinal surgery, to record the incidence of blood contamination of the hands, and to evaluate the protective effect of double gloving. DESIGN: Randomised prospective trial. SETTING: University hospital, Denmark. MATERIAL: 566 pairs of gloves tested on surgeons, assistants, and scrub nurses. INTERVENTIONS: The participants were allocated to wear either single gloves or Indicator double gloves. MAIN OUTCOME MEASURES: The number of glove perforations and the incidence of blood contamination of the hands. RESULTS: The perforation rate in single gloves was 53/306 (17%), and that of both outer and inner Indicator gloves at corresponding sites was 6/260 (2%) (p < 0.0001). Double gloving reduced the rate of blood contamination of the hands among surgeons from 15/115 (13%) to 2/98 (2%) (p < 0.005). CONCLUSION: The use of Indicator double gloves is recommended in gastrointestinal surgery because of the appreciable protection against blood contamination that they offer. PMID- 10817325 TI - Neither low-calorie diet nor vertical banded gastroplasty influence gastro oesophageal reflux in morbidly obese patients. AB - OBJECTIVE: Investigate the effect of a liquid diet, and of vertical banded gastroplasty, on gastro-oesophageal reflux in morbidly obese patients. DESIGN: Prospective study, patients being their own controls. SETTING: University hospital, Sweden. SUBJECTS: 25 morbidly obese patients, listed for vertical banded gastroplasty. INTERVENTIONS: 24 hour ambulatory intra-oesophageal pH monitoring three weeks before operation and repeated after 10-14 days on a liquid very low calorie diet, and finally three weeks after operation, while still on a liquid very low calorie diet. MAIN OUTCOME MEASURES: Percentage of time that pH in the oesophagus was < or =4. RESULTS: There were no changes in the amount of reflux, either after a liquid diet or after vertical banded gastroplasty. CONCLUSIONS: Neither the liquid diet nor vertical banded gastroplasty alone had any appreciable effect on gastro-oesophageal reflux. PMID- 10817326 TI - Tonic motor activity of the narrow gastric tube used as an oesophageal substitute. AB - OBJECTIVE: To assess the compliance, the existence of basal tone, the sensory response to distension, and the integrity of the neurohumoral enterogastric inhibitory reflex of the narrow gastric tube as an oesophageal substitute. DESIGN: Explanatory experimental study. SETTING: University hospital, The Netherlands. PATIENTS: Eight patients after oesophagectomy, formation of a narrow gastric tube, and cervical gastro-oesophagostomy. INTERVENTIONS: Measurements of intragastric tonic motor activity with a Barostat. MAIN OUTCOME MEASUREMENTS: Compliance, sensations during stepwise pressure increments, and changes in tone induced by glucagon and intraduodenal feeding. RESULTS: The compliance of the narrow gastric tube was found to be low (median 13.5 ml/mmHg (range 5-21)). Most of the patients perceived minimal sensation on distension. All patients had phasic tonic contractions during distension of the gastric tube. The median (range) relaxation after glucagon had been given intravenously was 14 ml (range 3 57) (p < 0.05). The tonic reaction to nutrients in the duodenum was not significant. CONCLUSIONS: The compliance of the narrow gastric tube is low. Postprandial symptoms are not caused by distension of the proximal part of the narrow gastric tube. Basal gastric tone and phasic activity were at least partially restored over time after operation. The enterogastric inhibitory reflex is eliminated by oesophageal resection and reconstruction of a gastric tube. PMID- 10817327 TI - Intestinal perforation by foreign bodies. AB - OBJECTIVE: To present our experience of patients who presented with intestinal perforation by foreign bodies to our hospital between 1980 and 1998. DESIGN: Retrospective study. SETTING: University hospital, Spain. SUBJECTS: 21 patients who presented with intestinal perforation by foreign bodies. MAIN OUTCOME MEASURES: Size of perforations, nature of foreign bodies, clinical symptoms, radiological findings, treatment and outcome. RESULTS: The most common location was the terminal ileum (n = 11), followed by the rectosigmoid junction (n = 5). The objects were mainly bone fragments and toothpicks. The diagnosis was generally made at operation, and only 4 of the 21 patients had signs of pneumoperitoneum on the preoperative abdominal film. The most common treatment was simple suture of the defect. CONCLUSIONS: Intestinal perforation by a foreign body is uncommon, and normally affects the ileocaecal and rectosigmoid regions, in which it is unusual to find pneumoperitoneum preoperatively. It must be considered in the differential diagnosis of such conditions as acute appendicitis and diverticulitis. PMID- 10817328 TI - Introduction of herniorraphy with mesh plug and patch. AB - OBJECTIVE: To evaluate the introduction of the Perfix mesh plug and patch system for inguinal hernia repair. DESIGN: Prospective consecutive follow-up study. SETTING: Teaching hospital, Sweden. SUBJECTS: 139 patients with 145 hernias who were operated on for inguinal hernia with the Perfix mesh plug and patch technique during 1997. MAIN OUTCOME MEASURES: Operating time, sick leave, time to full recovery, morbidity, recurrence rate. RESULTS: The median operating time was 35 minutes (range 15-105) and the mean follow-up was 9 months (range 4-13). Office workers required a mean of 7 days off work (range 0-43) and manual workers 15 days (range 0-90). Retired patients took 21 days (0-30) to recover fully, office workers 22 days (7-70), manual workers 30 days (7-90), students 34 days (0 60) and unemployed patients 60 days (21-150). There were 17 minor complications within 30 days and 2 recurrences during the follow up period. CONCLUSION: Herniorraphy with a mesh plug and patch can easily be introduced with good short term results. PMID- 10817329 TI - High recurrence rate 12 years after primary inguinal hernia repair. AB - OBJECTIVE: To find out the long term recurrence rate after repair of the inguinal ligament (Griffith) for inguinal hernia in one hospital. DESIGN: Retrospective study. SETTINGS: Teaching hospital, The Netherlands. SUBJECTS AND INTERVENTION: All patients who had had a Griffith repair for a primary inguinal hernia in 1985 were re-examined after at least 12 years by an independent examiner. RESULTS: Of the 102 patients included in the study, 45 patients had died and 17 could not be traced. Of the remaining 40 patients (45 hernias), 10 (22%) had developed recurrences. In 4 patients the asymptomatic hernia was discovered by the investigator. 11 further patients had developed a hernia on the opposite site resulting in a total of 16 patients (40%) with bilateral hernias. CONCLUSIONS: The long term recurrence rate of an inguinal hernia by reconstruction of the inguinal ligament is high and even higher when assessed by physical examination. The high recurrence rate and frequent bilateral recurrence might favour repairs with mesh reinforcement. PMID- 10817330 TI - Suspected acute appendicitis: is ultrasonography or computed tomography the preferred imaging technique? AB - OBJECTIVE: To compare the sensitivity and specificity of unenhanced spiral computed tomography (CT) and ultrasonography (US) in patients with suspected acute appendicitis. DESIGN: Prospective study. SETTING: University hospital, Germany. SUBJECTS: 120 consecutive patients with acute appendicitis as a differential diagnosis, whose clinical findings were not enough to make operation essential, but were too severe to send home. INTERVENTIONS: CT and US of the appendix. MAIN OUTCOME MEASURES: Sensitivity, specificity, and positive and negative predictive value. RESULTS: The results were correlated with surgical and histopathological findings at appendicectomy or clinical follow-up. 93 patients had acute appendicitis, 27 patients did not. The sensitivity of CT was 95% and of US 87%. The corresponding specificities were 89% and 74%, positive predictive values 97% and 92%, negative predictive values 83% and 63%. In the 27 patients who did not have acute appendicitis, the correct diagnosis was established with CT in 14 patients and with US in eight. CONCLUSION: CT is more sensitive and specific than US in patients suspected of having acute appendicitis, but in whom the presentation is equivocal. The use of unenhanced spiral CT led to a significant improvement in the accuracy of preoperative diagnosis and a lower negative appendicectomy rate. PMID- 10817331 TI - Vascularised knee joint transplantation in man: the first two years experience. AB - OBJECTIVE: To describe our early experience with a new technique for restoring destroyed knee joints to give reasonable functional results. DESIGN: Observational clinical trial. SETTING: Level-1-Trauma centre, Germany. SUBJECTS: 5 patients with large bone defects of the knee and loss of the extensor apparatus caused either by serious injury alone, or infection after serious injury. INTERVENTIONS: Transplantation of fresh and perfused knee joints with a vascular pedicle from multiorgan donors under immunosuppression. MAIN OUTCOME AND MEASURES: Ability to walk, need to remove one transplanted joint. RESULTS: Four patients are able to walk, the range of movement being from 50 degrees-120 degrees. The first patient additionally had to be provided with a total knee joint arthroplasty. In the third patient the graft became infected and had to be removed. She finally had an arthrodesis and bone lengthening by the Ilizarov technique. CONCLUSIONS: Transplantation of the knee joint may be an alternative to bone lengthening or amputation for patients with total loss of the extensor apparatus. PMID- 10817332 TI - Vascular structure and microcirculation of experimental pancreatic carcinoma in rats. AB - OBJECTIVE: To study angiogenesis and microcirculation in experimental pancreatic carcinoma. DESIGN: Open experimental study. SETTING: 2 University hospitals, Germany and USA. ANIMALS: 16 male Lewis rats. INTERVENTIONS: Induction of a duct like pancreatic cancer in the pancreas and peritoneum by interposition of fragments of tumour between 2 inert transparent polymethylmethacrylate plates. MAIN OUTCOME MEASURES: Microcirculation in the tumour and interaction between leucocytes, tumour, and endothelium investigated by intravital microscopy. RESULTS: The density of vessels in the carcinoma was significantly less than in normal pancreatic tissue (p = 0.0004). The vasculature of the tumour was characterised by a lack of differentiation in architecture of vessels, formation of sinusoidal and lacunar vessels and sudden changes in diameter of vessels. Red blood cell velocity differed among tumour vessels, but mean values were similar to those of normal exocrine pancreas. The mixed lymphocyte culture test indicated that the cell line DSL6A was immunogenic. However, high-affinity leucocyte endothelium-interaction was significantly reduced in the tumour's microcirculation after both orthotopic and heterotopic implantation (p = 0.002). Rates of apoptosis were suppressed in heterotopic tumours compared with orthotopic ones. Tumour growth was faster in heterotopic tumours. CONCLUSIONS: Experimental duct-like pancreatic carcinoma can be differentiated from normal pancreas by: chaotic arrangement of vessels with loss of differentiation of architecture and heterogeneous distribution; the formation of sinusoidal or lacunar vessels; lower vascular density with similar erythrocyte velocity; increase in mean diameter of vessels; reduced leucocyte-endothelium interaction despite confirmed immunogeneity independent of wall shear rates. The site of implantation influences apoptosis and growth rates. PMID- 10817333 TI - Acute pancreatitis and amaurosis after right parathyroidectomy for primary hyperparathyroidism. PMID- 10817334 TI - Treatment of an infected aortobifemoral graft by explantation and in situ reconstruction with an autogenous aortobifemoral saphenous vein graft. PMID- 10817336 TI - How to repair an adult indirect inguinal hernia? PMID- 10817335 TI - Acute mesenteric ischaemia: minimal invasive management by combined laparoscopy and percutaneous transluminal angioplasty. PMID- 10817337 TI - A novel perfusion system for animal cell cultures by two step sequential sedimentation. AB - A novel perfusion system was developed for high density culture of animal cells. The system consists of an airlift bioreactor, a setting tank and a flat settler. Both the settling tank and flat settler have two connecting pipes for transporting the cells from and back to the reactor, respectively. Thus, the cell flow in the settlers can be controlled in uni-direction, avoiding the countercurrent flow of the cells. During perfusion cultures, the cells firstly settled in the settling tank, then, unsettled cells in the tank were transferred to the flat settler for re-settling. With the application of the system to hybridoma cell cultures, it was found that the maximum viable cell density, monoclonal antibody concentration and average productivity were 1.31 x 107 cells ml-1, 400 mg l-1 and 461 mg l-1 d-1, respectively, which were much higher than those of a batch culture. Both theoretical analysis and experimental results showed a much higher separation efficiency in such a two-step sedimentation system than that in a conventional one-step sedimentation system. In addition, the volumetric ratio of the sedimentation devices to the culture volume in our developed system is much lower, which may be potentially useful on an industrial scale. PMID- 10817338 TI - D-malate production by permeabilized Pseudomonas pseudoalcaligenes; optimization of conversion and biocatalyst productivity. AB - For the development of a continuous process for the production of solid D-malate from a Ca-maleate suspension by permeabilized Pseudomonas pseudoalcaligenes, it is important to understand the effect of appropriate process parameters on the stability and activity of the biocatalyst. Previously, we quantified the effect of product (D-malate2 -) concentration on both the first-order biocatalyst inactivation rate and on the biocatalytic conversion rate. The effects of the remaining process parameters (ionic strength, and substrate and Ca2 + concentration) on biocatalyst activity are reported here. At (common) ionic strengths below 2 M, biocatalyst activity was unaffected. At high substrate concentrations, inhibition occurred. Ca2+ concentration did not affect biocatalyst activity. The kinetic parameters (both for conversion and inactivation) were determined as a function of temperature by fitting the complete kinetic model, featuring substrate inhibition, competitive product inhibition and first-order irreversible biocatalyst inactivation, at different temperatures simultaneously through three extended data sets of substrate concentration versus time. Temperature affected both the conversion and inactivation parameters. The final model was used to calculate the substrate and biocatalyst costs per mmol of product in a continuous system with biocatalyst replenishment and biocatalyst recycling. Despite the effect of temperature on each kinetic parameter separately, the overall effect of temperature on the costs was found to be negligible (between 293 and 308 K). Within pertinent ranges, the sum of the substrate and biocatalyst costs per mmol of product was calculated to decrease with the influent substrate concentration and the residence time. The sum of the costs showed a minimum as a function of the influent biocatalyst concentration. PMID- 10817339 TI - Influence of controlled glucose oscillations on a fed-batch process of recombinant Escherichia coli. AB - The influence of glucose oscillations on cell growth and product formation of a recombinant Escherichia coli culture producing a heterologous alpha-glucosidase was studied in fed-batch cultures in a laboratory bioreactor. Glucose oscillations were created by an on/off-feeding mode in either fast cycles (1 min) or slow cycles (4 min) and compared to a process with constant glucose addition. The study indicates that glucose oscillations influence the product stability and the overgrowth of plasmid-free cells if such cultures are not performed under continuous pressure for selection of plasmid-containing cells. Although the glucose uptake capacity decreased after induction of the recombinant alpha glucosidase in all cultures performed, the up-growth of plasmid-free cells during the production phase was strongly inhibited by fast oscillations. In contrast, plasmid-free cells grew up when constant feeding or slow cycles were applied. Our data suggest that the various feed protocols effect the specific carbon dioxide formation rate differently, with the highest production of carbon dioxide in the cultivations with fast cycles. In connection to product formation the initial alpha-glucosidase accumulation was the same in all cultures, but the stability of the product was significantly lower in the cultivation with slow cycles. Our results from laboratory experiments are discussed in relation to the mixing situation in large-scale bioreactors. PMID- 10817340 TI - Application of model-predictive control based on artificial neural networks to optimize the fed-batch process for riboflavin production. AB - The fed-batch process for commercial production of riboflavin (vitamin B2) was optimized on-line using model-predictive control based on artificial neural networks (ANNs). The information required for process models was extracted from both historical data and heuristic rules. After each cultivation the process model was readapted off-line to include the most recent process data. The control signal (feed rate), however, was optimized on-line at each sampling interval. An optimizer simulated variations in the control signal and assessed the forecasted model outputs according to an objective function. The optimum feed profile for increasing the product yield (YB2/S) and the amount of riboflavin at the time of harvesting was adjusted continuously and applied to the process. In contrast to the control by set-point profiles, the novel ANN-control is able to react on-line to variations in the process and also to incorporate the new process information continuously. As a result, both the total amount of riboflavin produced and the product yield increased systematically by more than 10% and the reproducibility of seven subsequently optimized batches was enhanced. PMID- 10817341 TI - Electrically stimulated induction of hsp70 gene expression in mouse astroglia and fibroblast cells. AB - Mouse astroglial cells, which were cultured on an electrode, were found responsive to an electric stimulation of sine wave potential in enhancing hsp70 mRNA resulting from an activation of hsp70 gene expression. On the basis of this finding, electrically responsive cells were established by transfecting mouse 3T3 L1 cells with a constructed plasmid encoding hsp70 promoter and the firefly luciferase gene. A stable cell line has been established through selection of heat-stimulated luciferase expression. A 1-h electric stimulation of the cells resulted in activation of luciferase expression, which was confirmed to produce an increase in light emission. The sequential pattern of the electrically stimulated expression of luciferase was found different from that of the heat stimulation. Furthermore, the promoter was activated depending on the potential and duration of the stimulation applied. Consequently, the electric stimulation has proven effective on activating hspP70 promoter. This cell line is feasible in expressing the gene of interest by electrical stimulation, which lead us to construct environment responsive cells in general. PMID- 10817342 TI - Use of a plasmid of a yersinia enterocolitica biogroup 1A strain for the construction of cloning vectors. AB - A plasmid with a size of 2,682 base pairs isolated from the Yersinia enterocolitica biogroup 1A strain # 29807 was characterized in respect to its suitability as a basic replicon for cloning vectors. The copy number of the plasmid was determined to be approximately 14 copies per cell and it was shown to be compatible with vectors with an origin of replication derived from ColE1 and p115A. The replication region of the plasmid encodes a primer RNAI and countertranscript RNAII. Two vectors, pIV1 and pIV2, containing a kanamycin resistance gene and the lacZalpha fragment with the multiple cloning site of pBluescriptSK + were constructed. A mobilizable derivative was successfully introduced into different bacteria belonging to the family Enterobacteriacea. To prove the applicability of the novel vectors for cloning purposes, a 13 kb hemolysin operon of Escherichia coli was inserted into pIV1, and the resulting recombinant plasmid was stably maintained and expressed in E. coli and Y. enterocolitica. PMID- 10817343 TI - Extent of incorporation of hydroxycinnamaldehydes into lignin in cinnamyl alcohol dehydrogenase-downregulated plants. AB - Down-regulation of cinnamyl alcohol dehydrogenase leads to an accumulation of cinnamaldehydes available for incorporation into the developing lignin polymer. Using electron spin resonance spectroscopy we have demonstrated that the parent radical of 4-hydroxy-3-methoxycinnamaldehyde is generated by peroxidase catalysed oxidation. The extent of radical generation is similar to that of 4-hydroxy-3 methoxycinnamyl alcohol and is increased by further aromatic methoxylation. From the distribution of the electron-spin density, it was predicted that the regiochemistry of 4-hydroxy-3-methoxycinnamaldehyde coupling would be similar to that of the corresponding alcohol, with the possibility of a higher degree of 8-O 4 linkages occurring. These predictions were confirmed by polymerisation studies, which also showed that after radical coupling the alpha,beta-enone structure was regenerated. This suggests that, although the cross-linking and physical properties of cinnamaldeyde rich lignins differ from that of normal lignins, cinnamaldehydes are incorporated into the lignin polymer under the same controlling factors as the cinnamyl alcohols. PMID- 10817344 TI - Determination of NADH-dependent glutamate synthase (GOGAT) in Spodoptera frugiperda (Sf9) insect cells by a selective 1H/15N NMR in vitro assay. AB - This is the second of two papers [Drews, M., Doverskog, M., Ohman, L., Chapman, B.E., Jacobsson, U., Kuchel, P.W., Haggstrom, L., 2000. Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR. J. Biotechnol. 78, 23-37]. where the general goal has been to determine and characterise the glutamine metabolism in Sf9 cells. The presence of glutamate synthase (GOGAT) activity was investigated in cell-free extracts of S. frugiperda (Sf9) insect cells by modified 1H/15N spin-echo and gradient enhanced multiple quantum coherence NMR spectroscopy techniques. Cell-free extracts were prepared from cells cultured in a serum-free medium. The assay conditions were based on conventional spectrophotometric and chromatographic methods. NMR data showed that nitrogen from [5-15N] glutamine was selectively incorporated into 2-oxoglutarate forming [2-15N] glutamate with a specific activity of 4.15 +/- 0.21 nmol [2-15N] glutamate min -1 (mg total protein)-1 in the cell-free extracts. The enzyme activity was exclusively dependent on NADH as coenzyme and was completely inhibited by 1 mM azaserine. From the results obtained, we conclude that Sf9 cells possess NADH-GOGAT activity. Furthermore, the high specificity of the NMR method enables distinction of competing reactions from glutaminase and glutamate dehydrogenase. PMID- 10817345 TI - Breast cancer screening uptake in women at increased risk of developing hereditary breast cancer. AB - This multicenter study assessed breast cancer screening uptake in 461 unaffected women at increased risk of developing breast cancer on the basis of family history who approached familial cancer clinics for advice about surveillance options. At the time of attending the clinic, 89% and 90% of participants were vigilant with respect to age- and risk-specific recommendations for mammography and clinical breast examination, respectively, and 51% reported practicing breast self-examination monthly or more frequently. The degree to which health outcomes are perceived to be under one's personal control (chi2 = -2.09, p = 0.0037) and breast cancer anxiety (chi2 = 8.11, p = 0.044) were both associated with monthly or more frequent breast self-examination, while there were no associations with sociodemographic characteristics. A significantly lower percentage (56%) of women aged <30 were vigilant with respect to mammography recommendations, compared to 77%, 96% and 98% of women aged 30-39, 40-49 and >50, respectively (chi2 = 37.2, p < 0.0001). These relatively low rates of mammographic screening in young women may reflect concerns about increased cancer risk associated with early and repeated radiation exposure or lack of sensitivity in young women with radiographically dense breasts. If mammographic screening is ultimately shown to lower mortality in women at high risk, there will be a strong case to promote screening in young women. The need for regular mammographic screening would then need to be highlighted and reinforced amongst young women and their referring physicians. Awareness amongst general practitioners, who are largely responsible for referral to screening services, would also need to be increased. PMID- 10817346 TI - The prognostic value of proliferation indices: a study with in vivo bromodeoxyuridine and Ki-67. AB - Proliferation indices are intended to help patients and clinicians make treatment decisions. We have previously demonstrated that a proliferation index based on in vivo labeling of S-phase cells with bromodeoxyuridine (BrdUrd) correlates with Ki 67 labeling index (LI). We now compare the prognostic value of these indices. With written consent, we gave 129 women with biopsy confirmed breast cancer 200 mg/M2 BrdUrd during 30 min immediately preceding surgery. We used IU-4 anti BrdUrd antibody to count the immunohistochemical labeling index (LI) of DNA incorporated BrdUrd in 2,000 cells and MIB-1 to count Ki-67 (118 cases). Patients received standard surgical and adjuvant treatment. No patients were lost to follow-up and patients were followed a minimum of 2 (median 5.1) years. We compared survival and recurrence in tumors with high vs low labeling indices. We found that women in the low BrdUrd LI group had better disease free survival (92% vs 67% 5-yr DFS p = 0.001) and overall survival (94% vs 70% 5-yr OS, p = 0.0001) than those with a high LI. In comparison, a low Ki-67 index predicted better OS (87% vs 80% 5-yr OS, p = 0.020) and a trend for better DFS (84% vs 72% DFS p = 0.055). The apparent superiority of BrdUrd LI over Ki-67 LI is likely due to chance (p = 0.18). In multivariate survival analyses we found that BrdUrd LI proliferative index significantly improves prediction of DFS or OS even when node status, age or tumor size is in the model. We conclude that markers of proliferation are useful adjuncts in predicting patient prognosis. PMID- 10817347 TI - Mammographic density changes in perimenopausal and postmenopausal women: is effect of hormone replacement therapy predictable? AB - BACKGROUND: Mammographic density adversely affects diagnostic accuracy and may be a risk factor for breast cancer. Mammographic density is affected by hormone replacement therapy (HRT). OBJECTIVE: To assess mammographic density in postmenopausal women with and without HRT. METHOD: Part I. Mammographic density was determined in 1232 postmenopausal women attending a breast screening clinic. Density was compared between HRT users and nonusers in three age groups. When available, previous mammograms were assessed for interval density change. Part II. Density change in 162 women during transition from premenopause to postmenopause was recorded. HRT effect was assessed. RESULTS: Part I. There was no density difference between postmenopausal HRT-users and nonusers younger than age 55. Thereafter, the density was significantly less in nonusers. In the majority of HRT-users the density remained at pretreatment levels but density increased in 8% of women after HRT was started. Part II. Of 117 HRT nonusers followed over menopause 38% had a density decrease before age 55. Of those started on HRT, 80% had no density change, 18% decreased density and 2% increased density. CONCLUSIONS: The density decreases significantly after age 55 with the greatest change occurring between age 55 and 64. This decrease does not occur in the majority of HRT-users. Usually HRT maintains the density present at the time HRT is started. Density decreases after HRT is started in some women suggesting refractoriness to hormones. In fewer than 8% of women density increases commensurate with HRT. Any adverse effect of HRT may depend on the receptivity of the epithelial elements which, in turn, may be reflected by the mammographic density at the time HRT is started. PMID- 10817348 TI - Single agent epirubicin as first line chemotherapy for metastatic breast cancer patients. AB - In order to better explore the toxicity and the activity of high dose epirubicin (120 mg/m2, 3 weeks) we analyzed a population of 127 metastatic breast cancer patients, treated in a randomized clinical trial conducted to evaluate the cardioprotective effect of dexrazoxane against epirubicin induced cardiotoxicity. All the patients had a diagnosis of metastatic breast cancer, an ECOG performance status < or = 2 and normal hematologic, renal, hepatic and cardiac function. No prior adjuvant chemotherapy including anthracycline was allowed. Epirubicin was given at the dose of 120 mg/m2 i.v. bolus every 3 weeks. One hundred twenty five patients were evaluable for toxicity and response. Seventeen patients (11%) had a complete response and 47 patients (37%) a partial response, for an overall response rate of 48%. The median progression free and overall survivals were 8.3 months and 18.3 months, respectively. Grade 3 and 4 leukopenia were observed in 8% and 7% of the patients, respectively. The most frequent nonhematological grade 3 toxicities were alopecia (87%), nausea and vomiting (16%), and mucositis (8%). Cardiotoxicity, defined as occurrence of congestive heart failure, decrease in resting left ventricular ejection fraction (L-VEF) to < or = 45%, or 20 EF units decrease from baseline L-VEF, was observed in 19% of the patients, after a median cumulative dose of epirubicin of 720 mg/m2 (range 120-1440). This study confirms in a large series of patients the activity of high dose epirubicin; however, the high incidence of cardiotoxicity requires a careful evaluation of cardiac risk factors before treatment. PMID- 10817349 TI - Infiltrating dendritic/Langerhans cells in primary breast cancer. AB - It is fully anticipated that dendritic cells (DCs) will become a mainstay for inclusion in biological therapies for patients with cancer including breast cancer. To elucidate the cellular composition of DCs infiltrating human breast cancers, we investigated the correlations between the density of infiltrating DCs and some clinicopathological factors of breast cancer patients, examined cytokine expression on cancer cells and finally, assessed the numbers of CD45RO+ tumor infiltrating lymphocytes (TIL). Tissues adjacent to cancer nests contained significantly more S-100 protein+ and S-100 protein+ CD1a- DCs, but less CD1a+ DCs, than the nests. In invasive ductal carcinomas infiltration by S-100 protein+ DCs within and adjacent to nests, CD1a+ DCs within nests and S-100 protein+ CD1a- DCs adjacent to nests was denser than that in non-invasive carcinomas. With respect to the histological subtypes, there were fewer DCs in scirrhous carcinomas. Patients with stage IV disease had significantly fewer DCs of primary lesions than at other clinical stages. There were good correlations between infiltration by S-100 protein+ DCs and expression of the cytokines GM-CSF, IL 1alpha and TNF-alpha on cancer cells and between GM-CSF expression and S-100 protein+ CD1a- DCs. There was a close correlation between CD45RO+ TIL and S-100 protein+ DC densities both within and adjacent to the cancer nests and the S-100 protein+ CD1a- DC density adjacent to the cancer nests. Despite extensive immunoelectron microscopic observation, CD1a+ DCs within cancer nests contained only few Birbeck's granule-like structure. These data indicate that cancer nests are infiltrated predominantly by CD1a+ DCs, whereas S-100 protein+ CD1a- DCs predominate in surrounding tissues, and a infiltration by DCs may require cytokine expression on cancer cells and simultaneous lymphocyte infiltration. The findings of this clinicopathological study indicate the importance of evaluating simultaneously the types and localizations of infiltrating DCs in cancer tissues. PMID- 10817350 TI - Shorter CAG repeat length in the androgen receptor gene is associated with more aggressive forms of breast cancer. AB - The androgen receptor (AR) is a transcription factor mediating the action of androgens. The AR gene is localized on chromosome X and it contains a series of CAG trinucleotide repeats. The length of the CAG repeats varies among individuals and this polymorphism is believed to be related to AR transcriptional activity. Studies have shown that fewer CAG repeats are associated with an increased risk as well as more aggressive forms of prostate cancer. Although AR is expressed in breast cancer and the impact of androgen and AR on breast cancer has been recognized, the role of the CAG repeats in breast cancer remains unknown. In this study, we measured the CAG repeats in breast cancer tissue using a PCR-based method. Of the 133 patients with primary breast cancer, 102 were heterozygous and 31 were homozygous. The mean CAG repeat number for homozygous women was 21; for heterozygous women the repeat number mean was 20 for the short allele and 24 for the long allele. The length of CAG repeats either in one allele or in both alleles was inversely correlated with the histological grade of breast cancer (r = -0.23 or -0.26, respectively, p < 0.05). An association between positive lymph nodes and fewer CAG repeats in both alleles was also suggested (p = 0.06). Furthermore, survival analysis indicated that the total number of CAG repeats in both alleles was associated with patient overall survival. With every CAG repeat increase, there was a 6% reduction in the risk of death (RR = 0.94, p = 0.03). The association remained significant after controlling for the homozygous and heterozygous status (RR = 0.92, p = 0.01). The association became no longer significant when clinical and pathological variables were adjusted in the analysis but this could be due to the reduction of sample size in the multivariate analysis. CAG heterozygosity and difference in number of CAG repeats between the two alleles were not associated with either disease features or patient survival. Our results suggest that longer CAG repeats may occur more frequently in less aggressive cancer and that the CAG repeats may play a role in breast cancer progression. PMID- 10817351 TI - Lipid peroxidation, free radical production and antioxidant status in breast cancer. AB - Reactive oxygen metabolites (ROMs), including superoxide anion (O2*-), hydrogen peroxide (H2O2) and hydroxyl radical (*OH), play an important role in carcinogenesis. There are some primary antioxidants such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) which protect against cellular and molecular damage caused by the ROMs. We conducted the present study to determine the rate of O2*- and H2O2 production, and concentration of malondialdehyde (MDA), as an index of lipid peroxidation, along with the SOD, GPx and CAT activities in 54 breast cancer (BC) patients. Forty-two age- and sex matched patients with minor surgical problems, who had no history of any neoplastic or breast disorders, were taken as controls. The rate of O2*- production was significantly higher (p < 0.001) in BC patients than controls, irrespective of clinical stages and menopausal status. Similarly, H2O2 production was significantly higher in BC patients, especially in stage III and postmenopausal groups, as compared to the respective controls. MDA concentration was also observed significantly elevated in stage II (p < 0.001), stage III (p < 0.01), postmenopausal (p < 0.005), and premenopausal (p < 0.02) group as compared to their corresponding controls. SOD and GPx activities were found significantly raised in all the groups (p < 0.001), except the GPx activity was found a smaller alteration in stage IV (p < 0.02). On the contrary, CAT activity was found significantly depressed in all the study groups. The maximum depression was observed in stage II (-61.8%). Lower CAT activity in our study may be the effect of higher production of ROMs, particularly O2*- and *OH. SOD and GPx, however, were less effected by these higher ROMs production. The results of our study have shown a higher ROMs production and decreased CAT activity, which support the oxidative stress hypothesis in carcinogenesis. The relatively higher SOD and GPx may be due to the response of increased ROMs production in the blood. However, the higher SOD and GPx activities may be inadequate to detoxify high levels of H2O2 into H2O leading to the formation of the most dangerous *OH radical followed by MDA. Therefore, administration of CAT may be helpful in the management of BC patients. However, further elaborate clinical studies are required to evaluate the role of such antioxidant enzymes in BC management. PMID- 10817352 TI - Prognostic relevance of cerbB2 expression following neoadjuvant chemotherapy in patients in a randomised trial of neoadjuvant versus adjuvant chemoendocrine therapy. AB - Recent advances in the detection and treatment of breast cancer have led to an intensive search for new markers of both prognosis and chemoresponsiveness. The oncogene cerbB2 has proved to be one of the most promising markers currently under study, both as a predictor of chemoresponsiveness and as a marker of poor prognosis. In addition the increasing use of neoadjuvant chemotherapy has led to the loss of standard prognostic criteria. In order to study the potential role of cerbB2 expression as an indicator of chemoendocrine resistance and poor prognosis, both before and after chemotherapy, we obtained tumour sections from 283 women enrolled onto a neoadjuvant trial. In this trial patients were randomised to receive either primary surgery followed by adjuvant chemoendocrine treatment or neoadjuvant chemoendocrine therapy followed by surgery. CerbB2 status was determined immunohistochemically on all of these patients. Thirty eight percent of the tumours were cerbB2 positive. There was no significant difference in expression between the adjuvant (41%) and neoadjuvant arms (35%). CerbB2 positive patients were much more likely to have shown non-response to chemoendocrine therapy (p < 0.001) and had a worse DES (p < 0.05). The best prognosis was seen in cerbB2 negative patients receiving neoadjuvant chemoendocrine therapy who showed a significantly better DFS (p < 0.05), than the cerbB2 negative patients receiving adjuvant therapy. PMID- 10817353 TI - Surgical treatment of hepatic metastases from breast cancer. AB - We have performed a retrospective study to evaluate whether surgical treatment is beneficial in patients with hepatic metastases from breast cancer. Between September 1985 and September 1998, 25 patients with hepatic metastases (14 solitary and 11 multiple), eight of whom had extrahepatic metastases, underwent hepatectomy. All of the detectable liver metastasis were resected in all of the cases. There were no severe postoperative complications. All but one of the patients received adjunctive polychemotherapy after the hepatectomy. After the hepatectomy, recurrent tumors were detected in 18 of the patients, being located in the liver in 12 (67%) of them. Overall, however, hepatectomy ensured that the liver was clinically recurrence-free for a median of 24 months (range 2-132 months). Eleven patients died of recurrent tumors, two died of other causes and the remaining 12 are currently alive. The 2- and 5-year cumulative survival rates after hepatectomy were 71% and 27%, respectively, and the median survival duration was 34.3 +/- 3.2 months, much better than the period of 8.5 months for another series of patients treated with standard or non-surgical therapies at our institution. The number and the size of hepatic metastases, the interval between treatment of the primary lesion and hepatectomy, and the existence of extrahepatic metastasis were not adverse prognostic factors. In conclusion, our data, although limited and highly selective, suggest that surgical treatment of hepatic metastases from breast cancer may prolong survival in certain subgroups of patients to a greater extent than standard or non-surgical therapies. PMID- 10817354 TI - Survival and tumor characteristics of German hereditary breast cancer patients. AB - Reports from different countries have been inconclusive in attempting to relate the BRCA1 mutation status to the survival of breast cancer patients. The purpose of this study was to investigate overall and disease-free survival for German hereditary breast cancer patients. Data on clinical outcome and data on age at diagnosis of breast cancer, histology, tumor size, lymph node status, histological grade, and laterality of 36 breast cancer patients from 12 families with a BRCA1 mutation and from one family with strong evidence for linkage to BRCA1 were compared with those of 49 hereditary breast cancer patients from 23 families that did not harbor a BRCA1 mutation. Overall and disease-free survival was estimated for both groups. BRCA1 mutation carriers had a significantly earlier age of diagnosis than non-carriers (p = 0.0001) and more frequently developed contralateral breast cancer (p = 0.04). Also, BRCA1-associated tumors more frequently were of larger size (p = 0.041) and higher grade of malignancy (p = 0.005) than non-BRCA1-associated tumors. Whereas no difference in overall survival was seen, disease-free survival at 10 years differed significantly with 53.3% for BRCA1 mutation carriers and 76% for non-carriers (p = 0.02). However, after stratification for age and in multivariate analysis for mutation status, age, and bilaterality, it was shown that the worse prognosis for BRCA1 mutation carriers disappeared. Our results suggest that the worse prognosis of BRCA1 mutation carriers in terms of disease-free survival may in large part be due to the age of onset of breast cancer in this population. Thus, BRCA1 mutation status does not appear to be an independent prognostic factor. PMID- 10817355 TI - Purification of an ascomycin derivative with simulated moving bed chromatography. A case study. AB - A purification process was developed for the separation of a semi-synthetic ascomycin derivative from its by-products. The process consists of a silica gel filtration and crystallization step prior to two simulated moving bed (SMB) separations, where in the first part the polar by-products and in the second part the apolar by-products were removed. The desired purity was achieved in a final crystallization step. Key elements of the whole process were the design of the first crystallization to obtain a product feasible for SMB chromatography and the specification of operating parameters for the two corresponding SMB steps. Starting from a crude product with an assay of only 44.9% an overall yield for the whole process of 81.0% was achieved with a final purity of >98%. PMID- 10817356 TI - Robustness testing of a reversed-phase high-performance liquid chromatographic assay: comparison of fractional and asymmetrical factorial designs. AB - Robustness tests were performed on a reversed-phase HPLC assay for triadimenol. Different experimental designs were compared. Two-level fractional factorial designs with different resolutions were used to study the influence of procedure related factors. The factors chromatographic column manufacturer at four levels and instrument at three levels were stepwise included in the study using asymmetrical factorial designs. The significance of the factor effects was determined statistically, using two types of error estimates in the calculation of critical effects, and graphically, by means of half-normal plots. The asymmetrical designs turned out to be an efficient and economic method to examine the influence of factors at different numbers of levels in the robustness testing of analytical methods. PMID- 10817357 TI - Solid-phase extraction followed by high-performance liquid chromatography ionspray interface-mass spectrometry for monitoring of herbicides in environmental water. AB - In this work we developed a sensitive and specific multiresidue method, based on reversed-phase liquid chromatography-mass spectrometry, with an ionspray interface (LC-ISI-MS), for determining 52 of most representative compounds of herbicides in water samples. The procedure used involved passing 0.5 l of surface water, 2 l of ground water and 4 l of drinking water samples, respectively, through a 0.5 g graphitized carbon black (GCB) extraction cartridge. Base-neutral and acid herbicides were differential eluted from GCB cartridge and follow analyzed by HPLC-ISI-MS apparatus. A conventional 4.6-mm-ID reversed-phase LC C18 column, operating with a mobile phase flow-rate of 1 ml/min, was used to chromatograph the analytes. A flow of 100 microl/min of the column effluent was diverted to the ISI source. The study demonstrates the sensitivity of the technique, with detection limit under 10 ng/l in drinking water samples. Performance data for the method such as recovery and precision are also reported. PMID- 10817358 TI - Liquid chromatography-electrospray ionization mass spectrometry of the diarrhetic shellfish-poisoning toxins okadaic acid, dinophysistoxin-1 and pectenotoxin-6 in bivalves. AB - Determination of diarrhetic shellfish-poisoning (DSP) toxins, okadaic acid (OA), dinophysistoxin-1 (DTX1) and pectenotoxin-6 (PTX6) was carried out by liquid chromatography (LC) followed by on-line atmospheric pressure electrospray ionization-mass spectrometric (ESI-MS) detection with a heated capillary interface. Mass spectra of authentic OA, DTXI and PTX6 standards exhibited abundant [M-H] at m/z 803, 817 and 887, respectively. Linearity of peak area obtained by selected-ion monitoring (SIM) for [M-H]- of each toxin was confirmed over a wide range of concentrations from 10 pg to 30 ng. LC-ESI-MS analysis of OA, DTX1 and PTX6 in scallops and mussels, collected at the same site (Mutsu Bay, Japan), was carried out. Scallops and mussels collected at the same site showed different toxin profiles. Although PTX6 was detected from scallops, it was not detected from mussels. PMID- 10817359 TI - Determination of anionic surfactants during wastewater recycling process by ion pair chromatography with suppressed conductivity detection. AB - A direct approach utilizing ion pairing reversed-phase chromatography coupled with suppressed conductivity detection was developed to monitor biodegradation of anionic surfactants during wastewater recycling through hydroponic plant growth systems and fixed-film bioreactors. Samples of hydroponic nutrient solution and bioreactor effluent with high concentrations (up to 120 mS electrical conductance) of inorganic ions can be analyzed without pretreatment or interference. The presence of non-ionic surfactants did not significantly affect the analysis. Dynamic linear ranges for tested surfactants [Igepon TC-42, ammonium lauryl sulfate, sodium laureth sulfate and sodium alkyl (C10-C16) ether sulfate] were 2 to approximately 500, 1 to approximately 500, 2.5 to approximately 550 and 3.0 to approximately 630 microg/ml, respectively. PMID- 10817360 TI - Extraction of hydrocarbon contamination from soils using accelerated solvent extraction. AB - Accelerated solvent extraction was studied as a method for the extraction of hydrocarbon contamination from wet and dry soils. Temperatures from 125 to 200 degrees C and six different solvents were investigated. Nonpolar solvents could not achieve complete recovery from wet soils at the temperatures studied. Optimum conditions were found to be 175 degrees C with dichloromethane-acetone (1:1, v/v) with 8 min heat-up time and 5 min static time. Quantitative recoveries for diesel range organics (DROs) and waste oil organics (WOOs) were obtained using the optimized conditions. The recovery of DROs and WOOs from three matrices at two concentrations (5 and 2000 mg/kg) averaged 115%. These results show that accelerated solvent extraction can generate results comparable to those obtained using Soxhlet or sonication. PMID- 10817361 TI - Determination of acetic acid in aqueous samples, by water-phase derivatisation, solid-phase microextraction and gas chromatography. AB - The direct derivatisation of acetic acid with n-hexyl chloroformate and with benzyl bromide in water was evaluated. With n-hexyl chloroformate, acetic acid did not give the n-hexyl acetate derivative, but the reaction of acetic acid with benzyl bromide in aqueous solution resulted in the formation of benzyl acetate. The derivatisation of acetic acid with benzyl bromide and the headspace solid phase microextraction (SPME) of benzyl acetate were optimised. Under optimum conditions, the limit of detection for acetic acid was 260 nM, and the relative standard deviation of the overall procedure at 1.10(-4) M acetic acid was 15.6% (n = 10). A linear response was obtained in the 1 x 10(-4) to 5 x 10(-6) M concentration range (R2 = 0.993, n = 6). Although Carbowax-divinylbenzene (CW DVB)-coated fibres exhibited a higher extraction capacity for benzyl acetate, polyacrylate (PA) was selected, because its mechanical stability was better than that of CW-DVB fibres. Moreover, the relative standard deviation of the SPME was better with PA (1.5%, n = 10 at 1 x 10(-5) M) than with CW-DVB-coated fibres (8.0%, n = 10 at 1 x 10(-5) M). Thus, a new analytical method for the quantitative determination of micromolar concentrations of acetic acid in the aqueous phase was developed. This method is based on water-phase derivatisation with benzyl bromide, headspace SPME with PA fibres and GC-FID. It was observed experimentally that benzyl alcohol formed by hydrolysis of the reagent affected the fibre-gas phase partitioning of benzyl acetate. PMID- 10817362 TI - Analysis of pitch deposits produced in kraft pulp mills using a totally chlorine free bleaching sequence. AB - Two organic deposits accumulated in a Kraft pulp mill during pulping of Eucalyptus globulus wood and throughout a TCF (totally chlorine free) bleaching sequence were characterized. One deposit was collected after cooking and an oxygen delignification stage while the other was collected after bleaching with hydrogen peroxide. The deposits were Soxhlet extracted with acetone, and the extracts redissolved in chloroform and subsequently analyzed by gas chromatography (GC) and GC-mass spectrometry (MS) using short and medium length high temperature capillary columns, respectively. On the other hand, the insoluble residues left after the acetone extraction were analyzed by Curie-point flash pyrolysis-GC-MS and by pyrolysis-methylation-GC-MS. The compounds identified in the deposits arise from the E. globulus wood lipophilic extractives that survive the pulping and bleaching processes. Triglycerides were completely hydrolyzed during the Kraft cooking and the fatty acids dissolved. Steroids (alcohols, hydrocarbons, ketones and esters) and waxes were the main components in the deposit collected after the oxygen delignification stage. After the bleaching with hydrogen peroxide, content of the waxes were reduced and fatty acids appeared. High amounts of fatty acids salts were also identified in the deposit collected after the oxygen stage, and in minor amounts in the deposit collected after hydrogen peroxide bleaching. In contrast, this deposit was mainly made up of high amounts of lignin-derived phenolic moieties. PMID- 10817363 TI - Determination of triazines in soil leachates by solid-phase microextraction coupled to gas chromatography-mass spectrometry. AB - A solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) method was developed for the evaluation of the leachability order of selected triazines (propazine, terbuthylazine, sebuthylazine, ametryn, prometryn and terbutryn) in soil/sediment samples (organic carbon content ranging from 0.19 to 0.42%), analysing fractions collected from a soil packed microcolumn elution experiments. The procedure is fast, simple, highly sensitive and solvent free. SPME-GC-MS was also employed for the quantitative determination of triazines in the soil leachate, since the method showed good recovery yield. Detection limits were always better than 1 ng ml(-1). The method was tested on a contaminated landfill top soil. Prometryn and ametryn were identified through their MS spectra and then quantified. Terbuthylazine was used to assess recovery. Results compared well with those obtained by solvent extraction followed by HPLC-UV detection. PMID- 10817364 TI - Extraction of DDT [1,1,1,-trichloro-2,2-bis(p-chlorophenyl)ethane] and its metabolites DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene] and DDD [1,1 dichloro-2,2-bis(p-chlorophenyl)-ethane]) from aged contaminated soil. AB - Pressurised liquid extraction (PLE) was used to extract DDT [1,1,1,-trichloro-2,2 bis(p-chlorophenyl)ethane] and its metabolites, DDD [1,1-dichloro-2,2-bis(p chlorophenyl)ethane] and DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene] from an aged, contaminated soil. Using three sequential static phases, PLE removed an equivalent quantity of DDT and its metabolites as Soxhlet extraction, in less time and with less solvent. Recovery was almost quantitative, implying appropriate sample work-up and manipulation. PMID- 10817365 TI - Automatic microgravimetric determination of fats in milk products by use of supercritical fluid extraction with on-line piezoelectric detection. AB - An on-line supercritical fluid extraction-piezoelectric detection system was developed and applied to the quantitative gravimetric determination of total fat in food samples (skimmed milk and cocoa). The proposed assembly provides all the advantages of an on-line system as regards automation, in addition to acceptable sensitivity and precision. Its strength lies in the design of the interface between the supercritical fluid extractor and the piezoelectric detector. Samples of skimmed milk and cocoa are weighed in the extraction thimble, previously loaded with I g of diatomaceous earth. A temperature of 100 degrees C and a CO2 fluid density of 0.60 mg/ml are used for extraction. The linear calibration range thus achieved is 0.005-0.07% w/w total fat, and the relative standard deviation is +/-2.3% (n=11; P=0.05). The throughput is six samples h(-1) (for the overall process). The proposed method was used to determine the total fat in food samples (milk, cocoa), the results being competitive with those of the Soxhlet methods for the same purpose. PMID- 10817366 TI - Determination of vitamins A and E in milk powder using supercritical fluid extraction for sample clean-up. AB - A method for the analysis of the natural contents of vitamins A and E in milk powder has been developed. The method utilises supercritical fluid extraction, a miniaturised alkaline saponification procedure and reversed-phase HPLC with UV detection. Modifications of the sample matrix, combinations of static and dynamic modes of extraction and effects of changes in extraction parameters such as temperature, flow-rate, time, collection solvent and collection temperature were studied to optimise the extraction efficiency and selectivity. Supercritical CO2 at 80 degrees C and 37 MPa, modified with 5% methanol and pumped at a flow-rate of 1.0 ml/min, gave recoveries of 99 and 96% for vitamins A and E, respectively, using a 15 min static followed by a 15 min dynamic extraction. The measurements gave a within-day RSD of 4% for both vitamin A and E, and between-day RSDs of 4 and 8% for vitamins A and E, respectively. PMID- 10817367 TI - Chiral separation of unmodified amino acids with non-aqueous capillary electrophoresis based on the ligand-exchange principle. AB - A ligand exchange mechanism in non-aqueous capillary electrophoresis was employed for the separation of eight unmodified amino acids using chiral complexes of copper(II) with L-proline and L-isoleucine. The electrophoretic medium consisted of 25 mM ammonium acetate and 1 M acetic acid in methanol. We were able to completely separate the enantiomeric pairs of each of the investigated racemic amino acids. We also report the optimization of the separation parameters, such as pH*, composition of the complex, and concentration of the complexing agents. PMID- 10817368 TI - Quantitative studies on the adsorption of proteins to the bare silica wall in capillary electrophoresis. II. Effects of adsorbed, neutral polymers on quenching the interaction. AB - A novel method is reported for quantifying protein adsorption to naked silica tubings and for assessing the efficacy of polymers added to the background electrolyte as dynamic wall modifiers. It consisted of flushing a fluorescently labelled protein (myoglobin) into a capillary equilibrated in Tris-acetate buffer, pH 5.0, until full saturation of the potential adsorbing sites. Desorption was then affected by electrophoretically driving sodium dodecyl sulphate micelles into the capillary from the cathodic reservoir: the peak of eluted material is quantified by using a dual laser beam instrument able to read the fluorescein isothiocyanate-derivatized myoglobin at 520 nm and the internal standard (sulphorodamine) at 630 nm. Four polymers have been assessed as potential quenchers of interaction of proteins with the silica wall: hydroxypropylmethylcellulose (HPMC, Mr = 1000000), hydroxyethylcellulose (HEC, Mr = 27000), poly(vinyl alcohol) (PVA, Mr = 49000) and short-chain poly(dimethylacrylamide) [poly(DMA)] (average Mr ca. 150000). HPMC, poly(DMA) and PVA were effective in the 0.005 to 0.02% (w/v) range, whereas HEC was active in the 0.1 to 0.8% concentration range. All polymers, however, except for poly(DMA), exhibited a rather poor performance in suppressing protein interactions with the siliceous surface, and could inhibit adsorption only by, at most, 50% (contrary to oligoamines which can quench such interactions by >90%). It is hypothesized that dynamically adsorbed polymers leave ample regions of the capillary inner surface unmasked, thus allowing strong interactions of proteins with the silica wall. This is also confirmed by the modest reduction of electroendoosmotic flow upon polymer adsorption, as compared with an untreated silica surface. Although poly(DMA) can inhibit protein adsorption by as much as 85%, its hydrophobic nature could in turn provide more adsorption sites for less hydrophilic proteins than myoglobin. PMID- 10817369 TI - Analysis of aliphatic alcohol ethoxylates in terms of alkyl and ethylene oxide chain lengths by reversed-phase liquid chromatography with evaporative light scattering detection. AB - Aliphatic alcohol ethoxylates are nonionic surfactants which are incorporated in many industrial formulations as complex mixtures of alkyl homologs and ethylene oxide oligomers. Determination of both the homolog and oligomer distributions is required for product control. The proposed method consisted of three reversed phase liquid chromatographic separation steps carried out on the same C18-bonded silica column. The first step was a preparative one: the sample mixture was fractionated according to the alkyl chain length without discrimination between ethylene oxide oligomers by using methanol-water eluent. The even homologs (EH) were collected together in a single fraction, the odd homologs (OH) in another. In the second and third steps, respectively, EH and OH fractions were separated according to the alkyl chain length and the number of ethylene oxide units simultaneously by changing the mobile phase composition to acetonitrile-water and by using evaporative light scattering detection. PMID- 10817370 TI - Separation and determination of rare earth elements by Dowex 2-X8 resin using sodium trimetaphosphate as elution agent. AB - The distribution coefficients of rare earth elements and thorium with Dowex 2-X8, 200-400 mesh, a strongly basic anion-exchange resin, have been determined regarding four different concentrations of sodium trimetaphosphate (3 x 10(-3), 5 x 10(-3), 7 x 10(-3) and 0.01 M). The separation of the rare earths and thorium obtained from an Australian monazite has been investigated by anion-exchange chromatography with sodium trimetaphosphate concentration gradient on a Dowex 2 X8 ion-exchange columns. The order of elution of the elements was the reverse of the order of elution of the same elements on Dowex 1 resin. The elution was investigated using 5 mg and 250 mg samples. In the separation of 5 mg samples, all elements were separated in 29 min. It has been seen that the elution peaks are narrow, tailing effects are very small, Dy and Y are well separated. Qualitative and quantitative determinations were realized by spectrofluorometry. PMID- 10817371 TI - Electric hypersensitivity and neurophysiological effects of cellular phones- facts or needless anxiety? PMID- 10817372 TI - Human brain activity during exposure to radiofrequency fields emitted by cellular phones. AB - OBJECTIVES: The aim of this study was to explore the possible influence of radiofrequency (RF) radiation exposure on human brain function. METHODS: The electroencephalographic (EEG) activity of 19 volunteers was quantitatively analyzed. Ten of the subjects were men (28-48 years of age) and 9 were women (32 57 years of age). The sources of exposure were 5 different cellular phones (analogue and digital models) operating at a frequency of 900 MHz or 1800 MHz. The EEG activity was recorded in an awake, closed-eyes situation. Six 30-minute experiments, including 1 sham exposure, were made for each subject. The duration of a real exposure phase was 20 minutes. RESULTS: Exposure to one of the phones caused a statistically significant change in the absolute power at the delta band of the EEG recording. However, no difference was seen in the relative power of the same band, and no changes occurred during exposure to other phones at any frequency bands. CONCLUSIONS: The findings of this study suggest that exposure to radiofrequency fields emitted by cellular phones has no abnormal effects on human EEG activity. The observed difference in 1 parameter was probably caused by statistical chance. PMID- 10817373 TI - Provocation of electric hypersensitivity under everyday conditions. AB - OBJECTIVES: In most previous provocation studies subjects suffering from "electric hypersensitivity" have not been able to determine correctly whether or not they have been subjected to a sham or true provocation to magnetic or electric fields. However, an often-discussed weakness is that most of the earlier provocation studies have been performed in a laboratory situation, often with simulated fields, which may not be representative of conditions prevailing in the homes or workplaces of the patients. Criticism has also been put forth about neglect of the long latency period of symptoms. Therefore, a provocation study was performed in the homes or workplaces of the patients, where we also studied the symptoms and on-off answer 24 hours after the exposure. METHODS: Fifteen subjects selected as having fast and distinct reactions from electric equipment were provoked on 4 occasions: mainly 2 true and 2 sham provocations. The intervals between exposure were a few or more days in order to provide the subjects with an opportunity to recover before the next provocation. A control group of healthy subjects with normal hearing and vision verified that the provocations were performed in a blind manner. RESULTS: The patients suffering from "electric hypersensitivity" were no better than the control group in deciding whether or not they were exposed to electric and magnetic fields. CONCLUSIONS: Exposure to electric and magnetic fields per se does not seem to be a sufficient cause of the symptoms experienced by this patient group. PMID- 10817374 TI - Cancer incidence among male pulp and paper workers in Norway. AB - OBJECTIVES: The study investigated cancer incidence among 23,718 male pulp and paper workers employed continuously for at least 1 year between 1920 and 1993 in Norway. METHODS: The name, date of birth, personal identification number, dates of hire and termination for all employment periods, specific department, and job categories were registered for each worker. Six subcohorts were established (sulfite mill, sulfate mill, paper mill, maintenance department, administrative staff and other departments). Data on the cohort were linked with data in the Norwegian Cancer Register. The follow-up period for cancer incidence, date of death, or emigration was from 1953 through 1993. RESULTS: An excess incidence of lung cancer was found among both short- and long-term employees [standardized incidence ratio (SIR) 1.5, 95% confidence interval (95% CI) 1.13-2.03 and SIR 1.2, 95% CI 1.09-1.34, respectively], especially for workers with the longest latency (SIR 1.3, 95% CI 1.08-1.44) and for sulfite mill workers (SIR 1.5, 95% CI 1.09-1.99). The risk for pleural mesothelioma was also increased (SIR 2.4, 95% CI 1.45-3.75), especially among maintenance workers. The results also showed an increased risk for malignant melanoma (SIR 1.3, 95% CI 1.04-1.60), an unexpected finding. CONCLUSIONS: Almost all the increased risk for lung cancer can be explained by a combination of smoking habits and asbestos use. although an effect of other work-related exposures (sulfur and chloride compounds, wood dust) cannot be excluded. Most of the cases of pleural mesothelioma occurred in departments where asbestos was used. There is no clear explanation for the excess of malignant melanoma, and the finding may be a chance occurrence. PMID- 10817375 TI - Cancer incidence among Norwegian airline pilots. AB - OBJECTIVES: In this retrospective cohort study, the cancer incidence of commercial pilots was studied to determine whether exposure at work has any influence on the incidence of cancer. METHODS: The cohort was established from the files of the Civil Aviation Administration and included people who had valid licenses as commercial pilots between 1946 and 1994. Basic data about their flight careers were recorded, and exposure to cosmic radiation was estimated. The cohort was linked to the Cancer Register of Norway. The observed number of cases was compared with that expected based on national rates. RESULTS: A group of 3701 male pilots was followed over 70 560 person-years. There were 200 cases of cancer versus 188.8 expected, with a standardized incidence ratio (SIR) of 1.06 and a 95% confidence interval (95% CI) of 0.92-1.22. No significant decreased risk was found for any cancer site. Excess risks were found for malignant melanoma (22 cases SIR 1.8, 95% CI 1.1-2.7) and nonmelanoma skin cancer (14 cases, SIR 2.4, 95% CI 1.3-4.0). For malignant melanoma, there was a significant trend for the SIR by cumulative dose. CONCLUSIONS: For most cancer sites, the incidence among pilots did not deviate from that of the general population and could not be related to block hours of flight time or dose. It seems more likely that the excess risks of malignant melanoma and skin cancer are explained by factors related to life-style rather than by conditions at work. PMID- 10817376 TI - Incidence rates of malignant mesothelioma in Denmark and predicted future number of cases among men. AB - OBJECTIVES: This study analyzed the incidence rates of malignant mesothelioma in Denmark in order to predict the future number of cases that will occur among Danish men. METHODS: The 1912 cases of malignant mesothelioma reported to the Danish Cancer registry in 1943-1993 were analyzed in order to describe current incidence rates. By a Poisson regression model the relative risks of synthetic birth cohorts were estimated and used in the prediction of the future number of cases that will occur among Danish men. RESULTS: The incidence rate increased to 1.33 per 100000 person-years in 1983-1987 among men and to 0.51 in 1973-1977 among women. From the Poisson regression model, the risk for birth cohorts of men, relative to the 1940-1944 cohort, peaked in the 1940-1944 cohort and decreased to 0.57 in the 1950-1954 cohort. The age-specific incidence rate peaked at 246 per 100000 person-years in the age group 80-84 years. The future annual number of mesothelioma cases is expected to peak around 2015 with 93 cases among men born before 1955. CONCLUSIONS: The fit of the models was not ideal, but with careful interpretation of the results, it was concluded that a further increase in the number of mesothelioma cases can be expected, and the effect of regulating the environmental exposure to asbestos cannot be expected within the next 10-15 years. PMID- 10817377 TI - Body burden of aluminum in relation to central nervous system function among metal inert-gas welders. AB - OBJECTIVES: The relationship between elevated internal aluminum loads and central nervous system function was studied among aluminum welders, and the threshold level for adverse effect was defined. METHODS: For 65 aluminum welders and 25 current mild steel welders body burden was estimated, and the aluminum concentrations in serum (S-Al) and urine (U-Al) were analyzed with graphite furnace atomic absorption spectrometry with Zeeman background correction. Referents and low-exposure and high-exposure groups were defined according to an aggregated measure of aluminum body burden, the group median S-Al levels being 0.08, 0.14, and 0.46 micromol/l, respectively, and the corresponding values for U Al being 0.4, 1.8, and 7.1 micromol/l. Central nervous system functions were assessed with a neuropsychological test battery, symptom and mood questionnaires, a visual and quantitative analysis of electroencephalography (EEG), and P3 event related potentials with pitch and duration paradigms. RESULTS: Subjective symptoms showed exposure-related increases in fatigue, mild depression, and memory and concentration problems. Neuropsychological testing revealed a circumscribed effect of aluminum, mainly in tasks demanding complex attention and the processing of information in the working memory system and in the analysis and recall of abstract visual patterns. The visual EEG analysis revealed pathological findings only for aluminum welders. Mild, diffuse abnormalities were found in 17% of the low-exposure group and 27% of the high-exposure group, and mild to moderate epileptiform abnormalities at a frequency of 7% and 17%, respectively. CONCLUSIONS: Both objective neurophysiological and neuropsychological measures and subjective symptomatology indicated mild but unequivocal findings dose-dependently associated with increased aluminum body burden. The study indicates that the body burden threshold for adverse effect approximates an U-Al value of 4-6 micromol/l and an S-Al value of 0.25-0.35 micromol/l among aluminum welders. PMID- 10817378 TI - Time to pregnancy among female greenhouse workers. AB - OBJECTIVES: This study examined the possibility that work in greenhouses with potential exposure to pesticides entails a risk for reduced fecundity in terms of increased time to pregnancy. METHODS: Among 1767 female members of the Danish Gardeners Trade Union, telephone interview data were obtained on the 492 most recent pregnancies of women employed when they stopped contraception to get a child (the starting time). The pregnancies were classified according to job characteristics at the starting time. The ratio between the likelihood of pregnancy during a month for the exposed persons versus the referents (the fecundability ratio) was estimated by discrete proportional hazards regression. RESULTS: The adjusted fecundability ratio for workers in flower greenhouses versus other union members was 1.11 [95% confidence interval (95% CI) 0.90-1.36]. Among workers in flower greenhouses the handling of cultures many hours per week, the spraying of pesticides, and the nonuse of gloves was related to reduced fecundability [adjusted fecundability ratio 0.69 (95% CI 0.47-1.03), 0.78 (95% CI 0.59-1.06), and 0.67 (95% CI 0.46-0.98), respectively]. CONCLUSIONS: The findings suggest that female workers in flower greenhouses may have reduced fecundability and that exposure to pesticides may be part of the causal chain. Additional studies of fertility among women working in greenhouses are highly warranted. PMID- 10817379 TI - Maternal occupational risk factors for oral clefts. Occupational Exposure and Congenital Malformation Working Group. AB - OBJECTIVES: This study investigated the role of maternal exposures at work during pregnancy in the occurrence of oral clefts. METHODS: The occupational exposures of 851 women (100 mothers of babies with oral clefts and 751 mothers of healthy referents) who worked during the first trimester of pregnancy were studied. All the women were part of a multicenter European case-referent study conducted using 6 congenital malformation registers between 1989 and 1992. In each center, the mother's occupational history, obtained from an interview, was reviewed by industrial hygienists who were blinded to the subject's status and who assessed the presence of chemicals and the probability of exposure. Odds ratios (OR) were estimated by a multivariate analysis including maternal occupation or occupational exposures during the first trimester of pregnancy and possible confounding factors such as center of recruitment, maternal age, urbanization, socioeconomic status, and country of origin. RESULTS: After adjustment for confounding factors, cleft palate only was significantly associated with maternal occupation in services such as hairdressing [OR 5.1, 95% confidence interval (95% CI) 1.0-26.0] and housekeeping (OR 2.8, 95% CI 1.1-7.2). The analysis suggests that the following occupational exposures are associated with orofacial clefts: aliphatic aldehydes (OR 2.1, 95% CI 0.8-5.9) and glycol ethers (OR 1.7, 95% CI 0.9-3.3) for cleft lip with or without cleft palate and lead compounds (OR 4.0, 95% CI 1.3-12.2), biocides (OR 2.5, 95% CI 1.0-6.0), antineoplastic drugs (OR 5.0, 95% CI 0.8-34.0), trichloroethylene (OR 6.7, 95% CI 0.9-49.7), and aliphatic acids (OR 6.0, 95% CI 1.5-22.8) for cleft palate only. CONCLUSIONS: Due to the limited number of subjects, these results must be interpreted with caution. However, they point out some chemicals already known or suspected as reproductive toxins. PMID- 10817380 TI - Occupational exposure to inhalative irritants and methacholine responsiveness. AB - OBJECTIVES: Occupational exposures to inhalative irritants have been associated with an increased reporting of respiratory symptoms in previous studies. Methacholine responsiveness represents a continuous measure of airway responsiveness. As such, it may be less subject to recall bias and more sensitive to detecting effects of occupational exposure on airways. Such effects may be stronger among atopic persons. The objective of the study was to examine the relationship between self-reports of occupational exposure to dusts, gases, vapors, aerosols, and fumes and methacholine responsiveness. METHODS: A sample was studied of never smokers (N=3044) chosen randomly from 8 areas in Switzerland. Atopy was defined as any positive skin test to 8 inhalative allergens. Nonspecific bronchial reactivity was tested using methacholine chloride and quantified by calculating the slope of the dose-response. RESULTS: The methacholine slopes were 19% [95% confidence interval (95% CI) 6-32] higher for never smokers with exposure to dusts, fumes, vapors, gases, or aerosols than for the unexposed group. When only atopic never smokers were examined. the increase was larger (37%, 95% CI 7-75), and for persons with >2 positive skin prick tests the effect was still higher (42%, 95% CI -1.5-104). Exposure to vapors and aerosols was strongly associated with increased methacholine slopes among the atopic subjects. CONCLUSIONS: Occupational exposure, particularly to dusts and fumes, was associated with increased bronchial reactivity in never smokers in this study. The magnitude of the effect was larger among atopic subjects. PMID- 10817381 TI - Modification of serum proteins in guinea pigs immunized and challenged with toluene diisocyanate. AB - OBJECTIVES: Guinea pigs were used to determine whether immunization and challenge by toluene diisocyanate (TDI) induce changes in the serum protein concentrations of the "acute-phase response" and whether TDI can form adducts with serum proteins. METHODS: Guinea pigs were immunized by weekly intradermal injections of TDI and challenged with TDI 7 days after the 3rd injection. The animals were killed 6 hours after the challenge, and serum was analyzed for protein characterization by gel electrophoresis and for specific antibodies to TDI by enzyme-linked immunosorbent assay (ELISA) and Western blotting. RESULTS: The total serum protein concentration of the immunized TDI-challenged guinea pigs increased in comparison with that of nonimmunized animals [75 (SE 0.7) versus 47.4 (SE 2.3) mg/ml; ]. Albumin and alpha, and alpha2 globulins increased significantly [respectively: 65.8 (SE 0.2)%, 2.1 (SE 0.1)% and 7.2 (SE 0.1)% versus 59 (SE 1.3)%, 1.3 (SE 0.1)% and 3.7 (SE 0.1)%], whereas beta1 and beta2 globulins decreased in the immunized TDI-challenged guinea pigs [7.8 (SE 0.2)% and 0.8 (SE 0.2)% versus 15.8 (SE 0.7)% and 4.8 (SE 0.2)%]. The gamma globulin concentrations did not change significantly. In the immunized TDI-challenged animals, albumin was modified by TDI and ran faster on agarose gel electrophoresis than did albumin from nonimmunized guinea pigs. In the ELISA, only immunized animals had high titers of TDI-specific antibodies (IgG and IgG1); by blotting, the antibodies reacted against TDI, the TDI-BSA-conjugate and several TDI-conjugated guinea pig serum proteins, but they did not react against any native or denaturated serum protein when unconjugated with TDI. CONCLUSIONS: These findings indicate that, in guinea pigs, immunization and challenge with TDI induces changes in serum proteins of the "acute phase response" and TDI is adducted to serum proteins with different molecular weights (eg, albumin). PMID- 10817382 TI - Physical work load between 1970 and 1993--did it change? AB - OBJECTIVES: This study investigates changes in self-reported and expert-evaluated physical work loads between 1970 and 1993 in relation to calendar year, birth cohort, and gender in an urban and suburban population sample (232 men and 252 women) born between 1935 and 1952. METHODS: A self-administered questionnaire was answered in 1993 concerning different aspects of physical work loads between 1970 and 1993. With the use of a classification matrix, the objective physical work load on different body regions was also assessed. RESULTS: Between 1970 and 1993 the fraction of subjects in blue-collar occupations and the physical work loads decreased among the men, but they both increased among the women. Physical work loads were, in general, higher among the men than among the women at younger ages (below 30 years), but less so at higher ages. Expert evaluations of the musculoskeletal load showed a pattern similar to that of self-reported work loads. CONCLUSIONS: The gender difference in work load development with age may have implications for the development of musculoskeletal disorders. PMID- 10817383 TI - Unemployment and health care utilization. AB - OBJECTIVES: This study attempted to determine whether prior use of health services predicts a subsequent risk of unemployment and also to describe the acute effects of exposure to unemployment on the use of health care services. MATERIAL AND METHODS: The 1986 census records were linked with comprehensive health care information for the period 1983-1989 for over 44629 randomly selected residents of Manitoba, Canada. All cause and cause-specific rates of hospital admission and ambulatory physician contacts were compared between 1498 unemployed and 18272 employed persons across 4 consecutive time periods related to the onset of unemployment. RESULTS: The adjusted rates of hospital admission and physician contacts were higher among the unemployed across all 4 periods. When persons with a history of mental health treatment were excluded, health care use in the period prior to the onset of unemployment was equivalent among the employed and unemployed. When a history of mental health treatment was controlled for, all cause and cause-specific health care use was elevated among the unemployed during the unemployment spell. CONCLUSIONS: Unemployed persons had increased hospitalization rates before their current spell of unemployment. Much of this difference was due to the subgroup with prior mental health treatment. For persons without prior mental health care, hospitalization increased after a period of unemployment. PMID- 10817384 TI - Functional foods--blurring the distinction between food and medicine. PMID- 10817385 TI - The vertebral artery: developmental pathology. AB - The vertebral artery (VA), whose embryogenesis is unique, different from that of any other vessel, is characterised by a great variety of malformations and anomalies. Some of the formers are truly pathological (that is symptomatic); the latter are just either anatomic or angiographic by chance findings. All of them should be kept in mind by the surgeon approaching the deep cervical and cranio spinal regions, as well as by the interventional radiologist. Width and length anomalies of the VA, tortuosity and kinking, course anomalies, duplication and fenestration, persistence of primitive arteries, anomalies of collateral branches are discussed in the light of a literature review. Other pathologies of the cervical VA (spontaneous aneurysms and arteriovenous fistulae) associated exclusively with genetic diseases as neurofibromatosis type 1 (NF1) and fibro muscular dysplasia (FMD) are also mentioned. PMID- 10817386 TI - Chemical sympathectomy in the relief of persistent sympathetic pain after carpal tunnel release. A case report. AB - Although carpal tunnel syndrome (CTS) is a common entrapment neuropathy, reflex sympathetic dystrophy is an extremely rare and devastating complication. In this report, a case of chemical sympathectomy in a patient who had undergone previous three surgical interventions elsewhere for unilateral CTS, with persistent sympathetic pain. PMID- 10817387 TI - Fatal basilar artery thrombosis after traumatic cervical facet dislocation. Case report. AB - The authors detail a rare case of basilar artery thrombosis in a patient with traumatic cervical spine facet dislocation. Although the patient's deficits could initially be explained by the spinal injury, deterioration to a "locked-in-state" could not. In addition to vertebral artery injuries, the basilar artery can also be indirectly involved in cases of cervical spine trauma. In the rare viable patient, immediate reduction of cervical spine dislocation may allow endovascular thrombolysis, if not otherwise contraindicated. PMID- 10817388 TI - Stretching (?) of the spinal cord as a cause of paraplegia in a patient with Cushing's syndrome. AB - Paraplegia with a total anesthesia level was found in a patient who had Cushing's syndrome. There was marked osteoporosis of the spine and a moderate wedge shaped compression fracture of the T4 vertebral body but no stenosis of the canal could be observed. On the grounds of the CT findings stretching of the spinal cord caused by the rapidly evolving kyphosis of the thoracic tract has been thought to be the responsible factor of the clinical deficits. PMID- 10817389 TI - Lymphocytic hypophysitis. Case report. AB - Lymphocytic hypophysitis is a very unusual disease typically observed in the peripartum period but found also in non-pregnant women or in men. We report the case of a 50-year-old woman with a five-year history of erithema nodosus for which was treated with variable doses of steroids. One year before admission the patient began to complain of headache, amenorrhea and rapidly progressive hypopituitarism. Magnetic resonance imaging showed an expanding sellar mass with homogeneous contrast enhancement while lacking the hyperintense signal of posterior lobe. The MRI findings and the history of autoimmune disease raised the suspicion of hypophysitis. The growth of the lesion and its unresponsiveness to the prolonged steroid therapy made surgery, which is both diagnostic and therapeutic, mandatory. The pathogenesis, diagnosis and management of this unusual clinical condition are discussed. PMID- 10817390 TI - Multiple meningiomas in different neuraxial compartments. Report of two cases. AB - Multiple meningiomas in different neuraxial compartments are quite rare. The authors report two new cases of association between cranial and spinal meningiomas, one of them in a patient operated upon for multiple intracranial meningiomas. The first case was a 60-year-old woman with progressive paraparesis who had been operated on 13 years earlier for multiple intracranial meningiomas. A myelo-CT scan showed a block of contrast medium at T1-T2; the lesion was removed via a standard laminectomy. The second patient was a 76-year-old woman with a 6-month history of spastic paraparesis. MRI detected an extramedullary intradural lesion at T6-T7. A cerebral MRI, performed because of the onset of seizures, showed a right parietal lesion. Removal of the thoracic lesion was followed, 6 months later, by removal of the cerebral one. Both patients presented a progressive improvement of the paraparesis and returned to normal daily activities. PMID- 10817391 TI - Ruptured intracranial dermoid cyst. AB - Intradural dermoids are rare congenital tumors representing approximately 0.05% of all intracranial lesions. These benign tumors have a typical appearance on CT and MR due to their lipid components. The complication caused by rupture are the spillage of the fatty material into the cerebrospinal fluid. We report a case of a ruptured dermoid cyst showing fat/fluid levels in both side ventricles and fatty material in the subarachnoid space on CT and MR-imaging and the follow-up over four years after incomplete resection of the tumor. PMID- 10817392 TI - Abducens nerve paresis as first symptom of trigeminal neurinoma. Report of two cases and review of the literature. AB - Two cases of neurinoma of the Gasserian ganglion with abducens nerve paresis as the first symptom are described and 27 cases culled from the literature are reviewed. The anatomo-surgical and clinical-radiological features of this pathology were re-assessed in an attempt to identify the pathogenetic mechanism responsible for a trigeminal neurinoma manifesting with VI cranial nerve palsy. PMID- 10817393 TI - Surgical treatment of penetrating orbito-cranial injuries. Case report. AB - Penetrating orbital injuries are not frequent but neither are they rare. The various diagnostic and therapeutic problems are related to the nature of the penetrating object, its velocity, shape and size as well as the possibility that it may be partially or wholly retained within the orbit. The authors present another case with unusual characteristics and discuss the strategies available for the best possible treatment of this traumatic pathology in the light of the published data. The patient in this case was a young man involved in a road accident who presented orbito-cerebral penetration caused by a metal rod with a protective plastic cap. Following the accident, the plastic cap (2.5x2 cm) was partially retained in the orbit. At initial clinical examination, damage appeared to be exclusively ophthalmological. Subsequent CT scan demonstrated the degree of intracerebral involvement. The damaged cerebral tissue was removed together with bone fragments via a bifrontal craniotomy, the foreign body was extracted and the dura repaired. Postoperative recovery was normal and there were no neuro ophthalmological deficits at long-term clinical assessment. Orbito-cranial penetration, which is generally associated with violent injuries caused by high velocity missiles, may not be suspected in traumas produced by low-velocity objects. Diagnostic orientation largely depends on precise knowledge of the traumatic event and the object responsible. When penetration is suspected and/or the object responsible is inadequately identified, a CT scan is indicated. The type of procedure to adopt for extraction, depends on the size and nature of the retained object. Although the possibility of non-surgical extraction has been described, surgical removal is the safest form of treatment in cases with extensive laceration and brain contusion. PMID- 10817394 TI - Measurement of brain natriuretic peptide in plasma samples and cardiac tissue extracts by means of an immunoradiometric assay method. AB - We evaluated the analytical characteristics and clinical usefulness of a commercial immunoradiometric assay (IRMA) kit for brain natriuretic peptide (BNP). Mean (+/-SD) plasma BNP concentrations measured in 129 normal subjects were 2.9+/-2.7 pmol/l (median 2.2 pmol/l; range 0.1-12.4 pmol/l). The mean (+/- SD) value observed in healthy men (2.1 +/- 2.0 pmol/l, n = 49) was significantly (p=0.0009) different to that found in women (3.4 +/- 2.9 pmol/l, n=80). A positive relationship (R=0.214, p=0.0174) was found between BNP values and age. In 65 patients with cardiac diseases, BNP levels increased with the progression of clinical severity of disease; patients with more severe disease [NYHA functional class III-IV, mean (+/- SD) BNP +/- 254 +/- 408 pmol/l, n=22] showed significantly (p<0.0001) increased values compared to patients with mild symptoms of disease (NYHA functional class I-II, mean (+/- SD) BNP=19.6 +/- 17.2 pmol/l, n=43). Furthermore, in 32 patients with chronic renal failure, greatly increased (p<0.0001) BNP values were found both before (mean +/- SD=88. 1+/- 111.1 pmol/l) and after haemodialysis (mean +/- SD=65.6 +/- 76.7 pmol/l), with a significant reduction after haemodialysis (p=0.0004) compared to pre-haemodialysis. The mean (+/- SD) BNP value found in atrial extracts collected during aorto-coronary bypass operations in 15 patients was 14.5 +/- 51.9 pmol/g of cardiac tissue. Moreover, the mean (+/- SD) tissue levels of BNP in 7 heart transplant recipients were 128.4 +/- 117.2 pmol/g of cardiac tissue in atrium, 68.4 +/- 76.7 pmol/g in ventricle, and 10.9 +/- 8.5 pmol/g in interventricular septum. Finally, BNP values found in cardiac tissues of two subjects collected at autopsy were considerably lower (on average 1/1000) than those observed in cardiac tissues of patients with cardiac diseases. The IRMA method for BNP determination evaluated in this study showed a good degree of sensitivity, precision and practicability. Therefore, this method should be a reliable tool for the measurement of plasma BNP levels for both experimental studies and routine assay. PMID- 10817395 TI - Presence of choloyl- and chenodeoxycholoyl-coenzyme A thioesterase activity in human liver. AB - In human liver homogenate the formation of bile acid-CoA thioesters is localized both to the microsomal fraction catalysed by an ATP-dependent synthetase and to the peroxisomal fraction catalysed by the thiolase in the last step of the beta oxidative cleavage of the 5beta-cholestanoyl side chain. The cytosolic bile acid CoA:amino acid N-acyltransferase catalyse the conjugation of the CoA-activated bile acids with taurine or glycine prior to secretion into bile. The formation of bile acid-CoA esters is considered the rate-limiting step in bile acid amidation. So far, a bile acid-CoA cleaving activity has not been assessed in the research of bile acid amidation in human liver. In this work, a bile acid-CoA cleaving activity has been demonstrated at a rate that may influence the concentration of bile acid-CoA thioesters, free bile acids and amidated bile acids within the hepatocyte. Recently, it was shown that free chenodeoxycholic acid, formed by the thioesterase, is the physiological ligand of the farnesoid X receptor. A multiorganelle distribution of the bile acid-CoA hydrolytic activity was found. In the postnuclear fraction of human liver homogenate, apparent Km and Vmax for the cleavage of choloyl-CoA were 7.7 x 10-5 mol/L and 3.6 nmol x mg-1 x min-1 respectively. The corresponding values for chenodeoxycholoyl-CoA cleavage were 7.1 x 10-5 mol/L and 4.8 nmol x mg-1 x min-1. Hydrolytic activities were detected in the microsomal and the peroxisomal fractions where the bile acid-CoA esters are formed as well as in cytosol housing the N-acyltransferase activity. Compared to the bile acid-CoA synthetase activities, the hydrolytic activities were considerably higher, both in the postnuclear fraction and in the microsomal fraction. The thioesterase activities were in the same range as detected for the N-acyltransferase activities both in the postnuclear fraction and in the cytosolic fraction. The mere presence of thioesterase in microsomes, peroxisomes and cytosol seems counterproductive to bile acid amidation. The thioesterases may have an indirect regulatory function on the bile acid synthesis and are important for the regulation of bile acid synthesis by providing free chenodeoxycholic acid, the most potent activator of the farnesoid X receptor. PMID- 10817396 TI - Vitamin D deficiency in Pakistani premenopausal women living in Norway is not associated with evidence of reduced skeletal strength. AB - In a cross-sectional, age-matched study, vitamin D status and skeletal status were compared between 26 Pakistani premenopausal women living in Oslo and 24 Norwegian women. The serum calcidiol concentration was significantly lower in Pakistani women (mean and 95% confidence interval (CI) 22 (16-27) nmol l-1 vs. 65 (55-74) nmol l-1, p < 0.001) and serum iPTH significantly higher (5.4 (4.1-6.8) pmol l-l vs. 3.4 (3.0-3.7) pmol l-1, p<0.05). No differences concerning bone mineral density (BMD) measured by DXA in the lumbar spine, femoral neck and total body or ultrasound parameters of the calcaneus were found between the Pakistani and Norwegian women. The Pakistani women were advised to take vitamin D supplementation, but after 1 year no significant changes were found in serum calcidiol or serum iPTH in 17 Pakistani women. Premenopausal Pakistani women living in Oslo, Norway seem to have increased risk of developing vitamin D deficiency with secondary hyperparathyroidism, but there is no evidence that their skeletal strength differs from that of Norwegian women. Advice on vitamin D supplementation did not result in a normalization of the vitamin D deficiency within 1 year of follow-up. PMID- 10817397 TI - Serum eosinophil cationic protein in asthma of farmers. AB - OBJECTIVE: There is need for relevant markers of bronchial inflammation in epidemiologic studies of asthma. Serum eosinophil cationic protein (ECP) is a marker of eosinophil inflammation and asthma activity. We have studied serum ECP in atopic farmers with current asthma, in non-atopic asthmatics and in non asthmatic, non-atopic controls. METHODS AND RESULTS: In a cross-sectional study of a representative sample of 8,482 farmers in Norway, asthma was recorded using a self-administered questionnaire; spirometry and serum sampling were performed on all of them. Atopy was screened with Phadiatop and RAST analyses to the mites Lepidoglyphus destructor and Tyrophagus putrescentiae in all asthma cases and controls. All the identified atopics had additional RAST analyses on a set of allergens. Serum ECP was tested in 60 persons with current asthma and atopy (mean 16.2, 95% CI 13.2-19.3), 127 non-atopic asthmatics (mean 9.1, 95% CI 8.0-10.2) and 39 non-atopic controls (mean 5.5, 95% CI 4.0 7.0). ECP levels in atopic asthmatics were associated with number of positive allergens and reduction of FEVI values. Moreover, the ECP levels were elevated with allergy to swine, cow, D. pteronyssinus, L. destructor, A. siro, T. putrescentiae, timothy grass and the cereal grains: wheat, oat, barley and rye. CONCLUSION: Serum ECP seems feasible as an indicator of inflammatory activity in epidemiological studies of current allergic asthma, and may help to indicate the importance of specific allergens. Although the ECP values were significantly more elevated in atopic than in non atopic asthma, elevated serum ECP was not specific for atopic asthma. PMID- 10817398 TI - Twenty-four hours of insulin infusion does not lower plasma lipoprotein(a) in healthy men. AB - The effect of 24-h exogenous hyperinsulinaemia on the plasma level of the atherogenic lipoprotein(a) (Lp(a)) is unknown. We evaluated the responses of plasma cholesterol, triglycerides, apolipoprotein (apo) B and Lp(a) during 24-h insulin infusion (180 pmol/kg/h) in 6 healthy men. Plasma total cholesterol (p < 0.01) and triglycerides (p < 0.05) decreased after 24 h of hyperinsulinaemia. Apo B was unchanged after 8 h (-2.4 +/- 3.0%, n.s.) and decreased by 10.9 +/- 4.8% (p<0.025) after 24 h of insulin. In contrast, Lp(a) did not decrease (+28.4 +/- 18.4%, n.s., and +/- 49.1 +/- 22.0%, n.s., after 8 and 24 h of insulin, respectively). This experiment supports the hypothesis that moderate hyperinsulinaemia has a different effect on the plasma level of triglyceride-rich lipoproteins compared to Lp(a). PMID- 10817399 TI - Impact of blood sampling in very preterm infants. AB - In a prospective investigation, 99 very preterm infants (gestational age (GA) 24 32 weeks, birthweight 560-2,255 g) were studied during the first 4 weeks of life. The infants were divided into two groups: infants born extremely early (GA <28 weeks, n = 20) and infants of GA 28 - 32 weeks; the groups were then subdivided into critically ill or not. Diagnostic blood sampling and blood transfusion events were recorded. In total, 1905 blood samples (5,253 analysis) were performed, corresponding to 0.7 samples (1.9 analysis) per day per infant. The highest frequencies were found during the first week, in infants with extremely low GA and in critically ill infants. The mean blood loss and transfusion volume values were 13.6 ml/kg and 6.3 ml/kg, respectively. In total, 19 infants (19%) received 34 transfusions corresponding to 0.3 transfusions per infant. Thirteen out of 20 infants of extremely low GA received 28 blood transfusions, corresponding to 27.0 ml/kg of blood on average during the study period. Four developed late anaemia; thus, in total, 14 (70%) of the infants born extremely early received 35 transfusions during the first 3 months of life, corresponding to a total mean of 34.8 ml/kg. For the extremely preterm infants a significant correlation between sampled and transfused blood volume was found (mean 37.1 and 33.3 ml/kg, respectively, r = + 0.71, p = 0.0003). The most frequently requested analyses were glucose, sodium and potassium. Few blood gas analyses were requested (1.9/ infant). No blood losses attributable to excessive generous sampling were detected. The results show an acceptable low frequency of sampling and transfusion events for infants of GA 28-32 weeks. The study emphasizes the necessity of thorough reflection and monitoring of blood losses when ordering blood sampling in extremely preterm, critically ill infants. PMID- 10817400 TI - Plasma concentrations of endothelial vasoactive substances in clinically healthy subjects. associations with urinary albumin excretion and ambulatory blood pressure. AB - Elevated urinary albumin excretion (UAE) is a predictor of cardiovascular disease, and one possible explanation is that elevated UAE reflects a generalized vascular dysfunction. The present study tests whether the plasma concentrations of the two main endothelial vasoactive substances (nitric oxide and endothelin-1 [ET-1]) are changed in clinically healthy subjects with elevated UAE (>6.6 microg/ min-the 90th percentile in the background population) and to test associations between these concentrations and systemic blood pressure. Twenty seven subjects with elevated UAE were compared with 46 matched controls with normoalbuminuria. Plasma concentration of ET-1 was measured using an ELISA method and plasma concentration of nitrate/nitrite using a photometric method. Twenty four-hour blood pressure was measured using a portable recorder (TM-2421). No significant differences in the concentrations of nitrate/nitrite and ET-1 were found between the groups, e.g. 21 (10-105) vs. 18 (11 -152) (p=0.33) and 0.98 (0.58 1.95) vs. 1.10 (0.54 -1.50) (p = 0.27), respectively. However, plasma nitrate/nitrite was significantly positively correlated to systolic and diastolic blood pressure in subjects with normoalbuminuria but not in subjects with elevated UAE. In contrast, plasma ET-1 concentration was significantly positively correlated to systolic blood pressure only in subjects with elevated UAE. In conclusion, elevated UAE is not associated with changed plasma concentrations of endothelial vasoactive substances in clinically healthy subjects. However, nitrate/nitrite is positively correlated to BP only in subjects with normoalbuminuria, and ET-1 is positively correlated to BP only in subjects with elevated UAE. PMID- 10817401 TI - Effects of SCH 42495, a neutral endopeptidase inhibitor, on cardiac function and mass in rats after repeated hyperbaric exposures. AB - The aim of the present study was to investigate the effects of inhibition of neutral endopeptidase on left ventricular hypertrophy (LVH), myocardial necrosis and haemodynamic changes, previously shown in rats after repeated hyperbaric exposures to 5 bar, by using the novel neutral endopeptidase inhibitor SCH 42495. Ten test rats underwent chamber dives daily for 40 consecutive days, and 10 control rats were exposed in the same chamber for an equivalent period of time, but in air at 1 bar. The rats received SCH 42495 orally (30 mg/kg twice a day) for 40 days. After 40 days, the body mass was identical in the two groups. Test rats and control rats had equal heart mass (both totally and left ventricular myocardium, including the ventricular septum/100 g), thus indicating that long term treatment with SCH 42495 inhibits hyperbarically induced LVH. The left ventricular pressure (LVP) and the maximal velocity of LVP increase and decrease (+/- dP/dt) were similar in the test rats compared to the control rats at 1 bar. Previously, we found a higher LVP and dP/dt in non-treated test rats in otherwise identical experiments. This indicates that SCH 42495 "normalizes" the cardiac function of test rats after repeated hyperbaric exposures. The systolic arterial pressure, heart rate (HR) and respiratory frequency (RF) were similar in the two groups throughout the experiments. However, treatment with SCH 42495 lowered the blood pressure compared to previously non-treated rats. In conclusion, long-term administration of the neutral endopeptidase inhibitor SCH 42495 prevented previously shown changes in cardiac function as well as myocardial mass after 40 consecutive exposures to 5 bar. PMID- 10817402 TI - Plasma levels of N-terminal proatrial natriuretic peptide in children are dependent on renal function and age. AB - Plasma levels of natriuretic peptides are used as diagnostic markers of heart failure. The aim of this study was to analyse the relation between plasma levels of N-terminal proatrial natriuretic peptide (Nt-proANP) and renal function, and to develop reference values in children. Nt-proANP was measured in the plasma of 86 patients whose glomerular filtration rate (GFR) was determined by use of the X ray contrast medium iohexol and a fluorescence technique. Blood samples for Nt proANP were also collected in 399 reference children, aged 0 - 15 years. The relationship between Nt-proANP and GFR was examined using a multiple regression analysis. The mean value of Nt-proANP was markedly higher in children with heart failure than in children with malignant or urologic diseases (p<0.001). The variability in plasma levels of Nt-proANP was mainly (adjusted R2=0.81) explained by the following four variables: presence of heart failure, GFR, age and previous treatment with anthracyclins. Plasma levels of the peptide are raised at birth, but fall rapidly to adult levels. We conclude that the plasma levels of Nt-proANP are age-dependent. Moderately elevated values were registered in children with severe renal impairment. Heart failure is regularly associated with excessive elevation of Nt-proANP in plasma. Our findings suggest that the influence of heart failure on levels of this peptide in children greatly exceeds the influence of renal dysfunction. PMID- 10817403 TI - Effect of cycloheximide on tryptophan binding to rat hepatic nuclei. AB - This study evaluated whether cycloheximide, an inhibitor of protein synthesis, would affect the binding of L-tryptophan to rat hepatic nuclei or nuclear envelopes. Previous reports have indicated that the binding of L-tryptophan to hepatic nuclear envelope protein was saturable, stereospecific, and of high affinity. Also, the administration of L-tryptophan rapidly stimulated hepatic protein synthesis. In this study, we determined that the addition of cycloheximide in vitro inhibited 3H-tryptophan binding to hepatic nuclei or nuclear envelopes. Heat-treated cycloheximide failed to have this inhibitory binding effect. In vivo treatment of rats with cycloheximide diminished in vitro 3H-tryptophan binding to hepatic nuclei of treated rats compared to controls. Puromycin, another inhibitor of hepatic protein synthesis, when added in vitro did not affect 3H-tryptophan binding to hepatic nuclei but did diminish in vitro binding after in vivo treatment. Thus, cycloheximide added in vitro diminished 3H tryptophan binding to hepatic nuclei probably by its structural effect on the receptor while cycloheximide administered in vivo may also act in part by inhibiting protein synthesis. PMID- 10817404 TI - Synthesis of conformationally constrained hydroxy-alpha-amino acids by intramolecular conjugate addition. AB - An efficient and easily applicable method for the synthesis of a variety of hydroxy-alpha-amino acids analogues of serine and phenylalanine has been established. The method involves the stereoselective intramolecular conjugate addition of the benzamide group to cyclohexenone promoted by Lewis acid. Subsequent transformations of functional groups provide the conformationally constrained 2-hydroxy- and 2,4-dihydroxy-6-phenylcyclohexane-alpha-amino acids. PMID- 10817405 TI - Polyamines are unevenly distributed within the rat and rabbit kidney. AB - Aliphatic polyamines have generally been measured on the whole kidney. Since the kidney is composed of a variety of cells, whole organ data are of limited value for the interpretation of the functions of the polyamines. The aim of this study was to establish the distribution pattern of putrescine, spermidine and spermine within the kidneys of male and female rats and rabbits. It is shown that the polyamines are unevenly distributed along the cortico-papillary axis. Each amine exhibited its own distinct distribution pattern. The polyamines are predominantly located in the cortex. Putrescine levels increased gradually from the cortex to the papillary tip in rabbits, whereas, in rats, fluctuations in putrescine level were marked. In the six zones of the rabbit kidney studied, spermidine and spermine concentrations were markedly higher in females than in males. This difference was less marked in rats. PMID- 10817406 TI - Homocysteine, vitamins B6, B12, folate, and risk of coronary artery disease in patients undergoing diagnostic coronary angiography. AB - Homocysteine and vitamins B were correlated with coronary artery disease in patients undergoing diagnostic coronary angiography. 160 patients having > or =1 stenosis (G1), 55 patients having normal coronary arteries (G2) and 171 healthy volunteers (G3) were prospectively recruited. Homocysteine levels were significantly higher in patients, particularly in those with normal coronary angiograms, than in healthy subjects (13.8 +/-6.3 micromol/L in G1 (p < 0.0001) and 15.2 +/- 8.8 micromol/L in G2 (p < 0.0001) versus 10.1 +/- 3.1 micromol/L in G3). Homocysteine levels were not related to the extent of coronary artery disease. In patients with normal angiogram, vitamin B12 and folate levels were significantly higher compared with the other groups (p < 0.05 and p < 0.001, respectively) showing that vitamin B deficiency was not involved in the hyperhomocysteinemia. In conclusion, homocysteine and vitamins B levels do not contribute to discriminate for the presence of coronary artery disease in patients undergoing diagnostic coronary angiography. Homocysteine levels, however, were higher in patients referred for coronary angiography than in healthy controls. PMID- 10817407 TI - Effects of hypoxia on plasma amino acids of fetal sheep. AB - Secondary amino acid disturbances from circulatory responses during hypoxia may cause problems in interpreting plasma amino acid profiles of sick babies investigated for possible inherited defects. Systematic studies to characterise them are difficult in man. We investigated the effects of hypoxia on plasma amino acids by studying 9 late gestation fetal sheep in utero during 11 one hour episodes of moderately severe isocapnic hypoxia. In 6 experiments, maternal plasma amino acids were also monitored. Fourteen fetal plasma amino acids increased significantly, with the largest proportionate changes in alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, ornithine and lysine. Maternal amino acids did not increase. Probable explanations were reflex peripheral vasoconstriction in skeletal muscle beds and decreased hepatic blood flow. The findings extend our knowledge of the fetal response to hypoxic stress, demonstrate the importance of skeletal muscle in branched-chain amino acid metabolism, and should help with interpretation of postnatal plasma amino acid disturbances. PMID- 10817408 TI - Importance of proline and other amino acids during honeybee flight--Apis mellifera carnica POLLMANN). AB - The levels of proline and other amino acids in the haemolymph and other body parts of honeybee foragers were investigated by HPLC analysis. The concentrations of proline in the blood of glucose-fed or -injected bees finishing their exhaustive tethered flights on a roundabout were significantly reduced compared to bees that were fed and rested for one hour. This indicates some utilization of proline during flight metabolism. The levels of essential amino acids and of the sum of all amino acids except proline remained roughly constant, indicating that the decrease of proline did not result from a changed haemolymph volume. 14C labelled proline was injected into bees either shortly before starting their flight or before a resting period of equal duration in an incubator at the same temperature. Bees that rested had incorporated more proline into thorax body protein, and less of the labelled substance was unrecovered ("missing") and considered to be respired or less probably defecated. If the entire amount of missing 14C-proline is regarded as exhaled, the oxidative breakdown of proline reached higher levels after flight than in rested bees. This is another hint that proline is utilized during flight. Usually the exhaled amount did not exceed 10 microg proline in half an hour of flight. Although our data indicate involvement of proline in flight metabolism, the amount metabolized is low compared to the utilization of carbohydrates. PMID- 10817409 TI - Synthesis and antibacterial activity of some new non-proteinogenic amino acids containing thiazole residues. AB - Some new thioamides and thiazoles have been synthesized using canavanine, S cysteine, homo-S-cysteinesulfonamides and their N-omega aminoethylated derivatives as adducts in order to investigate the structure-antimicrobial activity relationships. The compounds showed substantial antibacterial activity in vitro against various gram-positive (Staphylococcus aureus, Bacillus cereus etc.) and gram-negative (Escherichia coli, Proteus vulgaris etc.) bacteria. These findings indicate that the presence of the thiazole residue is an essential factor for the antibacterial effect. PMID- 10817410 TI - Conservation of the basic pattern of cellular amino acid composition during biological evolution in plants. AB - The cellular amino acid composition of plant cells was analyzed. The callus of carrot (Daucus carota), leaves of Torenia fournieri and protocomb-like body of Cymbidium, s.p. were examined as examples of plant cells. The cellular amino acid compositions differed in the plant cells, but their basic patterns were quite similar. It is concluded that the basic pattern of the cellular amino acid composition is conserved in all terrestrial organisms, including plants. PMID- 10817411 TI - Relationship between lifting capacity and anthropometric measures. AB - STUDY DESIGN: Prospective analysis of relationship between lifting capacity and multiple anthropometric variables. OBJECTIVES: To determine the relationship between lifting capacity and anthropometric variables and to model this relationship quantitatively. BACKGROUND: Low-back injuries commonly occur in individuals who perform lifting tasks. Objective data are needed to determine preinjury lifting capacity that, in turn, might be used to guide decisions during rehabilitation of these injuries. METHODS AND MEASURES: We recorded age and sex and measured the following variables for 35 men and 23 women between the ages of 22 and 40: height, weight, percentage of body fat, torso height, pelvic width, pelvic girth, arm length, thigh girth, and calf girth. Variables were selected for the study on the basis of theoretical modeling or previous research regarding the relationship between study variables and lifting capacity. Subjects also were tested to determine their maximum lifting capacity by using a lordotic lumbar spine lifting technique. RESULTS: Stepwise regression analysis indicated that the combination of sex, age, thigh girth, pelvic girth, and percentage body fat was significantly related to maximum lift capacity (multiple R2 = 0.76). The mean absolute difference (+/- SD) between lifted amount predicted by the regression model and the actual amount lifted was 118.6 +/- 86 N (26 +/- 19.3 lb), which corresponded to an average absolute error of 16% (SD = 14%) of the actual weight lifted. CONCLUSION: The results may be useful in estimating 1 aspect of preinjury lifting capacity. Similar studies are needed to model the requirements of frequency of lift, duration of lifting efforts, variety of hand-object coupling, and combined lifting and reaching. PMID- 10817412 TI - Metabolic and cardiorespiratory responses to continuous box lifting and lowering in nonimpaired subjects. AB - STUDY DESIGN: A within-subject experimental design. OBJECTIVES: To compare the magnitude of metabolic and cardiorespiratory changes produced during box lifting and lowering among combinations of lift technique (leg lift and leg-torso lift) and lift weight (10.8 and 15.4 kg). BACKGROUND: Continuous box lifting and lowering can be used as an exercise in a low-back rehabilitation program. Awareness of the possible cardiovascular stress of this activity is important to the clinician because some patients may have existing cardiovascular pathologies or possess unknown risk factors for cardiovascular disease. METHODS AND MEASURES: A group of 17 nonimpaired men 26 +/- 8 years of age (mean +/- SD) performed the 4 experimental trials on different days in a counterbalanced order determined by a Latin Square design. Lifting and lowering was performed for 6 continuous minutes at a rate of 12 cycles per minute. Physiologic variables were oxygen uptake, minute ventilation, heart rate, systolic blood pressure, diastolic blood pressure, metabolic equivalent, and rate-pressure product. RESULTS: There were stepwise increases in the values for oxygen uptake, minute ventilation, heart rate, metabolic equivalent, and rate-pressure product from the leg-torso lift to the leg lift and from 10.8 to 15.4 kg of weight within each lift technique (with the exception that minute ventilation and heart rate did not differ between the leg-torso lift at 15.4 kg and the leg lift at 10.8 kg). For the 4 lifts, values (mean +/- SD) varied from 20.3 +/- 5.4 to 28.8 +/- 5.8 mL x kg x min(-1) for oxygen uptake, 42.2 +/- 11.1 to 66.4 +/- 15.2 L x min(-2) for minute ventilation,129 +/- 20.6 to 156 +/- 16.5 beats x min(-1) for heart rate, 5.8 +/- 1.6 to 8.2 +/- 1.6 for metabolic equivalent, and 197 +/- 49.4 to 245 +/- 41.2 for rate-pressure product (x10(-2)). CONCLUSION: The leg lift with the 15.4-kg weight produced the greatest physiologic stress. Because of the magnitude of the increase in the variables measured for all 4 types of lifts, clinicians should closely monitor patients' response to this type of exercise. PMID- 10817413 TI - Transmissivity of coupling agents used to deliver ultrasound through indirect methods. PMID- 10817414 TI - Influence of Q-angle on lower-extremity running kinematics. AB - STUDY DESIGN: Two-group posttest-only comparison. OBJECTIVE: To assess the influence of the Q-angle on the 3-dimensional lower-extremity kinematics during running. BACKGROUND: An excessive Q-angle has been implicated in the development of knee injuries by altering the lower-extremity locomotion kinematics. Previous investigations using 2-dimensional analyses during walking did not support this hypothesis. METHODS AND MEASURES: We hypothesized that individuals with Q-angles more than 15 degrees would display an increase in rearfoot eversion and tibial internal rotation during running. Thirty-two nonimpaired subjects (men: n = 16, mean age = 22 +/- 3 years; women: n = 16, mean age = 23 +/- 3 years) ran over ground, and 3-dimensional kinematic data were collected from the right lower extremity. Subjects with a Q-angle of 15 degrees or less comprised the low-Q angle group, whereas those with Q-angles of more than 15 degrees comprised the high-Q-angle group. Segment and joint maximum angles and the times when the maxima occurred during stance were measured. RESULTS: The Q-angle magnitude did not increase the maximum segment or joint angles during running. The groups displayed similar maximum angles for rearfoot eversion (low Q-angle, -15.5 +/- 5.0 degrees; high Q-angle, -15.6 +/- 6.6 degrees) and tibial internal rotation (low Q-angle, -8.8 +/- 4.8 degrees; high Q-angle, -6.8 +/- 5.1 degrees). The high Q-angle group (39.5 +/- 16.3%) achieved maximum tibial internal rotation later in the stance phase than the low-Q-angle group (28.8 +/- 10.7%). CONCLUSIONS: In support of the previous investigations involving Q-angle influences on kinematics, our study did not reveal any differences between groups in maximum joint or segment angles. The kinematic information did reveal that the high-Q angle group displayed an increase in time to maximum tibial internal rotation. The impact of this single factor on producing knee injury is unknown. PMID- 10817415 TI - Measurements of temporal aspects of gait obtained with a multimemory stopwatch in persons with gait impairments. AB - STUDY DESIGN: Repeated measures of 14 temporal factors of gait obtained with a multimemory stopwatch from a variety of subjects with locomotor impairments. OBJECTIVES: To estimate the intratester and intertester reliability of 14 temporal factors of gait by using a multimemory stopwatch; to compare novice and expert clinicians at mastery of making these temporal measurements. BACKGROUND: Temporal gait measures are useful for describing the effectiveness of treatment interventions in patients with locomotor impairments. METHODS AND MEASURES: Eleven adult subjects (mean age, 48.4 years; SD, 5.7 years), 10 with locomotor impairments and 1 elderly adult, ambulated along a 6-m walkway 3 times at a self selected walking speed. The subjects were videotaped from the side as they walked. Four physical therapists independently analyzed the videotapes on 2 occasions; 2 examiners were recent graduates, and 2 others had 23 years of clinical experience. Intraclass correlation coefficients were used to estimate intratester reliability. A component of variance analysis quantified the sources of variation. RESULTS: Intraclass correlation coefficients for each of the 14 variables varied from 0.88 to 0.98. The major contributor to variance was subject, followed by trial, error, and tester; the tester factor generally contributed less than 1% to the total variance. CONCLUSIONS: Reliable measurements of the temporal aspects of gait can be made by using a multimemory stopwatch and videotape in a clinical setting on patients with various locomotor problems. Our data suggest that measurements obtained by more experienced physical therapists were no more reliable than those made by recent graduates. PMID- 10817416 TI - The social validation of three physical restraint procedures: a comparison of young people and professional groups. AB - The treatment acceptability of three forms of physical restraint was evaluated with three groups of raters. One restraint method involved personal restraint in a chair, two other methods involved personal restraint on the floor. The three groups of raters were special education staff, residential staff, and a group of young adults with no experience of residential services. Ratings, of videotaped role-play using the Treatment Evaluation Inventory (Kazdin, 1980) revealed that the chair method of restraint was rated as more acceptable than the other method to all three groups. The residential staff rated the chair method as more acceptable than the other methods. The results are discussed in terms of the importance of evaluating restrictive, emergency procedures, and future methodological refinements. PMID- 10817417 TI - Reducing overselective attention to compound visual cues with extended training in adolescents with severe mental retardation. AB - Because of the devastating impact a disturbance in responding to multiple cues can have on a child's development, this investigation determined whether computer touch-screen technology could be utilized to improve the attentional skills of students with severe developmental disabilities after attentional deficits were initially identified. In particular, we assessed whether establishing prior reinforcement histories for separate stimuli would control how adolescents with severe mental retardation attended to visual compounds when extended training was given. Initially, prior reinforcement contingencies of individual stimuli failed to control the attention of the adolescents (Huguenin, 1997). Longer single stimulus pretraining and additional exposure to compounds containing stimulus components with conflicting reinforcement histories, however, eventually proved effective in determining what aspects of complex visual cues they attended to. In most instances, the adolescents selectively responded to stimulus elements whose prior reinforcement histories were unchanged in the compound after additional training was administered. Stimulus elements with a reversed prior reinforcement contingency were usually ignored. The reliability of the effect of prior reinforcement histories of individual stimulus elements on attention to visual compounds following additional training was confirmed with multiple testing procedures, automatically administered by a computer. Even though presenting conflict compounds initially identified students with overselective attention, extended exposure to single stimulus training and conflict compounds alleviated stimulus overselectivity and improved their attentional skills. After individual stimulus-response relations were reestablished and sufficiently reinforced to reduce disrupting effects when compound training cues were presented, stimulus overselectivity was eliminated. Through longer single stimulus pretraining and additional exposure to training compounds, adolescents with severe mental retardation learned to selectively attend to each component of visual compounds when prior reinforcement histories associated with the individual stimulus elements were manipulated. The findings of this investigation indicated that overselective attention among students with developmental disabilities is not an unmodifiable perceptual characteristic. They also revealed that overselective attention may be due to the disrupting effects of compound training cues which can be minimized through longer single stimulus pretraining and repeated presentations of compound training cues. Utilizing computer technology to administer procedures similar to those described in this study may permit students with developmental disabilities to acquire essential attentional skills for learning educational tasks involving complex cues. PMID- 10817418 TI - Self-injurious behavior and sleep disturbance in adults with developmental disabilities. AB - Sleep patterns of 30 individuals with self-injurious behavior and mental retardation were compared with those of 30 matched controls residing in the same residential facility that did not self-injure. Individuals were recorded as asleep or awake during 30 min intervals for eight hours per night. The results of a Wilcoxon signed-ranks test (p < .05) indicated that individuals with self injury slept significantly less than individuals without self-injury. chi2 analyses (p < .01) indicated significantly greater variability in the number of intervals recorded as asleep among individuals with self-injury than their matched controls. These results are congruent with previous findings of sleep disturbance among persons with mental retardation and behavior problems. The possibility of neurochemical dysregulation in sleep disturbance among individuals with daytime self-injury is discussed. PMID- 10817419 TI - Response covariation of escape-maintained aberrant behavior correlated with sleep deprivation. AB - We examined the relation between sleep deprivation and changes in level and allocation of aberrant behavior (aggression and self-injury) for a child with moderate mental retardation. First, a series of functional analyses identified that self-injury (SIB) and aggression were maintained by escape from demands (i.e., were members of the same response class). Escape from demand conditions were then held constant over time while natural levels of sleep deprivation occurred. This final analysis demonstrated a correlation between sleep deprivation and increases in escape-maintained aberrant behavior. Sleep deprivation seemed to be related to increases in levels of self-injury but did not seem to influence levels of aggression. PMID- 10817420 TI - Effects of verbal elaborations on memory for sentences in adults with mental retardation. AB - We examined the effects of verbal elaborations on memory for verbal material in adults with mental retardation and a control group of adults of normal intelligence. During acquisition, three types of sentences were presented that differed in elaboration of the subject-adjective relationship: a) non-elaborated base sentences; b) base sentences with arbitrary verbal elaborations; and c) base sentences with explanatory verbal elaborations that clarified the significance of the subject-adjective relationship. At test, we varied the retrieval context by presenting either base sentences alone or base sentences with the verbal elaborations. A three-alternative, forced-choice recognition procedure was used to test memory for the target adjectives. Results yielded a positive effect of retrieval support (elaborations present at study and test) on recognition performance for adults with mental retardation. These data imply that the retrieval context plays an important role in recognition memory for adults with mental retardation. PMID- 10817421 TI - Teaching children with autism to initiate and sustain cooperative play. AB - This study investigated the acquisition, transfer, and maintenance of cooperative play in six children with autism. Two approaches were compared. In one approach, the participants observed two models engaging in cooperative play, before taking the place of one of the models while the play episode just observed was repeated. The second approach was identical to the first except that the participants were now required to verbally describe the modeled play episode before taking the place of one of the models. During training, modeled play episodes varied across play topics, and the criterion for mastery was first trial learning of novel play episodes. A nonconcurrent multiple baseline design across participants was applied. The results showed that the participants failed to acquire cooperative play until the verbal description was included in the training procedure. Following training with verbal description, all participants: a) could initiate episodes and sustain episodes initiated by their play partner; b) were able to take turns in episodes that were considerably longer than the episodes practiced during training; c) varied their play within and between play episodes; and, d) transferred those skills across play partners, settings, and time. PMID- 10817422 TI - The evolution of ductus arteriosus treatment. AB - The treatment of patent ductus arteriosus (PDA) has evolved over the years. We reviewed 231 non-premature children (group 1) undergoing surgical closure of a PDA between January 1985 and December 1997, and 30 children (group 2) undergoing transcatheter closure from May 1995 to December 1998. The median age and weight at operation in group 1 were 13 months (range, 0.5-174 months) and 9.5 kg (range, 1.9-49.7 kg), respectively. There was one intra-operative death (0.4%) secondary to hemorrhage. Immediate extubation was performed in 208 patients (90%). Intra operative chest tube use decreased from 73.3% to 10% between the 1985-88 and 1996 97 periods (P < 0.001). Postoperative pneumothoraces occurred in 33/131 (25%) patients with only one patient (0.7%) requiring drainage. Eleven patients had complications including wound infection in four, vocal cord paralysis in three, and left pulmonary artery stenosis in one. The median length of stay (LOS) was 5 days (range, 2-43 days). Follow-up echocardiogram was performed in 146/230 patients (63%) and revealed a residual PDA in six (4%); two being re-ligated, two remaining clinically insignificant, and two spontaneously resolved at 7 and 28 months follow-up. The remaining 84 patients had no clinical signs of a residual PDA. In group 2, where a transcatheter coil occlusion technique was used, the median age and weight at procedure were 31 months (range, 9-320 months) and 14.9 kg (range, 9-69.7 kg), respectively. Vascular complications occurred in four patients (13.3%). One patient developed hemolysis and hemoglobinuria requiring hospital admission. Four patients required a second intervention. At the most recent echocardiographic assessment, four patients (13.3%) had a residual PDA. PMID- 10817423 TI - The role of video-assisted thoracic surgery for the treatment of lung cancer: lung lobectomy by thoracoscopy versus the standard thoracotomy approach. AB - Our objective was to evaluate the usefulness, safety, validity and benefits of video-assisted thoracoscopic surgery (VATS) for performing pulmonary lobectomy in 24 patients with clinical NO stage I primary non-small-cell lung cancer compared with 30 patients who underwent a conventional thoracotomy. There were no significant differences in the intra-operative blood loss, duration of operation, or duration of chest tube drainage between the VATS group and the standard lobectomy group, but in this VATS' experience, patients had less postoperative pain. Numbers and distributions of dissected lymph-nodes were similar in patients whether undergoing standard thoracotomy or VATS lobectomy. We can confirm that the safety and validity of VATS are virtually identical to those of the standard thoracotomy approach in the lobectomy. However, the former technique causes less discomfort to patients and requires a shorter recovery period of laboratory data and IL-6 concentrations in thoracic drainage fluid. We conclude that VATS major lung resection is technically feasible. Stringent patient selection is important and special training is needed. PMID- 10817424 TI - Long-term effects of combined iliac dilatation and distal arterial surgery. AB - PURPOSE: When standard aortofemoral surgical procedure is combined with lower extremity vascular surgery, problems related with the hospital stay, morbidity, mortality and the cost of treatment will exist. The number of reports relating to combined iliac artery PTA and distal bypass surgery is limited. After the development of stenting procedures, the results of arterial system plasty have much more improved. This report reviews our preliminary experience with iliac artery angioplasty with distal bypass procedures. PATIENTS AND METHODS: A total of 41 patients have undergone combined iliac artery dilatation and distal arterial revascularization. Angioplastic procedures were performed in the angiography suite and distal surgery was carried out at the same day or the day after. Of all patients, 29 underwent percutaneous transluminal angioplasty (PTA) and 12 underwent combined PTA and stent placement. Ipsilateral femoropopliteal bypass was performed as a distal revascularization procedure in all patients. RESULTS: Mean systolic iliac artery pressure gradients improved from 34.7+/-8.6 mmHg to 3.9+/-3.2 mmHg after angioplastic procedures (P < 0.0001). Six patients needed reangioplasty because of restenosis in the follow-up period. Thrombectomy was performed on 1 patient in the early postoperative period and re-do femoropopliteal bypass was performed on two patients in the 2nd and 23rd months. Three minor wound infections were successfully treated with antibiotics and local care. Mean follow-up was 21.4 months (range 1-48 months). By life-table analyses, the overall 4-year cumulative primary patency of combined procedures was 78.1%. CONCLUSION: The results show that the combined procedure is a suitable method for the treatment of patients with multiple stenotic lesions at the iliac and distal arterial levels. We believe that the combined use of PTA and distal vascular surgery by an experienced surgical team will give beneficial results and a highly satisfactory outcome in this group of patients. PMID- 10817425 TI - Adrenalin tolerance does not prevent bacterial translocation in a murine burn model. AB - Burn injury causes mesenteric vasoconstriction and bacterial translocation. Since catecholamines are powerful vasoconstrictors and elevated immediately after burn injury, we hypothesised that adrenaline tolerance might decrease burn-induced mesenteric vasoconstriction and bacterial translocation. Adrenaline tolerance was developed in Swiss albino mice. Adrenaline tolerant and control animals were subdivided into sham-burn and burn subgroups. 24 h after sham-burn or burn injury, specimens were obtained for microbiological evaluation. Also, in a separate group of adrenaline tolerant and control animals, superior mesenteric blood flow was measured. Burn injury increased bacterial translocation rate in both control (P = 0.001) and adrenaline tolerant groups (P = 0.0351). The caecal bacterial level increase was significant after burn injury in control groups (P = 0.0004) but was not significant in adrenaline tolerant animals (P = 0.743). Mesenteric blood flow was decreased significantly by burn injury in both control and adrenaline tolerant animals (P < 0.00001). The results showed that catecholamines do not mediate postburn mesenteric vasoconstriction or bacterial translocation. PMID- 10817426 TI - The implication of nitric oxide in the process of bacterial translocation. AB - Since there is increasing evidence indicating nitric oxide [NO] would play a role in sepsis, we decided to investigate whether this multifaceted mediator is directly implicated in the process of bacterial translocation. A total of 48 rats received intraperitoneal either Zymosan A (group Z) for systemic inflammation production or sodium chloride solution (controls); they were then further subdivided into three groups of eight animals each, being given, through the tail vein: L-NAME (N-nitro-L-arginine] for inhibition of NO production; SNP (sodium nitroprusside) as NO donor; or sodium chloride as control. After 2 h, the mesenteric lymph node complex was excised, under sterile conditions, and, using standard bacteriological techniques, bacterial translocation was assessed as colony forming units per gram of tissue (CFU/g). Statistical evaluation of the bacteriological data revealed a significant increase of bacterial translocation in all rats subjected to systemic inflammation (group Z) versus controls (P = 0.01) Control rats that were subjected to L-NAME treatment exhibited a statistically significant increase (P = 0.001) in CFU/g compared to sodium chloride treated rats, while SNP treatment revealed no difference in relation to sodium chloride treated rats. Group Z rats, subjected to L-NAME treatment, similarly exhibited a statistically significant increase (P = 0.01) in CFU/g compared to sodium chloride treated rats, while SNP treatment led to a statistical increase of bacterial translocation in relation to sodium chloride treated rats (P = 0.05). The results of this study lead us to suggest that NO appears to participate in the process of bacterial translocation. PMID- 10817427 TI - Effect of plaunotol on bacterial translocation in the rat small intestine. AB - BACKGROUND AND AIMS: Bacterial translocation is precipitated by an increase in bacteria or endotoxin, depression of the membrane barrier, and an increase in mucosal permeability. Plaunotol is a mucosal protective agent, and observed to have a strong suppressive effect on superoxide production. In this study, the effect of plaunotol on bacterial translocation was examined using the model of ischemia and reperfusion. METHOD: Male Sprague Dawley rats were used to create the following model for evaluation of bacterial translocation: (i) the control group; (ii) the preventive dose group (plaunotol 30 mg/kg/day one week before surgery); (iii) the therapeutic dose group (plaunotol 30 mg/kg/day one week after surgery); and (iv) the full dose group (plaunotol 30 mg/kg/day one week before surgery and one week after surgery). Bacterial translocation was assessed as the blood concentration of the endotoxin. RESULTS: In the control group, the endotoxin increased significantly 3 days postsurgery (13.7+/-5.6 pg/ml) compared with before surgery (1.1+/-0.1 pg/ml). In the preventive and full-dose groups, the erndotoxin decreased significantly 3 days postsurgery (4.4+/-2.8 pg/ml, 5.7+/ 2.7 pg/ml, respectively) compared with that of the control group. CONCLUSION: Plaunotol in the preventive and full-dose groups decreased the endotoxin. This suggests that plaunotol is one of the protectors for bacterial translocation. PMID- 10817428 TI - Differences in gut integrity following abdominal surgery according to the magnitude of the surgical stress. AB - BACKGROUND: An increase in serum diamine oxidase (DAO) activity reflects intestinal mucosal damage. This was used to estimate the effect of surgical stress after elective abdominal surgery on gut integrity. METHODS: Patients with gastrointestinal tract cancers were placed in either group T (transthoracic esophagectomy for esophageal cancer, n = 9) or group L (laparotomy, n = 10). The serum DAO activity was measured pre-operatively, and on postoperative days 1, 3, and 8. RESULTS: Transthoracic esophagectomy was associated with a more extensive stress than the operations in group L, as measured by the intra-operative blood loss, the amount of blood transfused, and the operative time (P = 0.007, P = 0.0002, P = 0.0011, respectively). Following surgery, the serum DAO activity was decreased markedly in all patients in group T. In contrast, the activity was unchanged in group L (P = 0.04). CONCLUSIONS: The severity of the surgical insult plays a significant role in the decrease in serum DAO activity. These results suggest that surgical stress influences gut integrity following elective abdominal surgery. PMID- 10817429 TI - Sigmoidometry: a method assessing sigmoid colon function. AB - The purpose of this communication is to present a method that assesses the sigmoid colon (SC) volume, pressure and compliance both quantitatively and qualitatively. The response of the SC pressure to SC distension was recorded in 12 healthy volunteers. The SC was distended by means of a condom, which was applied to an 8 F tube and inflated with carbon dioxide at a rate of 150 ml/min. The SC pressure was measured by a saline-perfused 8 F tube and the intra abdominal pressure by a catheter placed in the urinary bladder. A curve called 'sigmoidometrogram' could be recorded and assessed qualitatively and quantitatively. The curve had a 'tone limb' representing the SC pressure upon distension and an 'evacuation limb' representing the SC pressure during balloon expulsion. The former showed insignificant pressure elevation (P > 0.05) with increasing volumes of CO2 inflation. It exhibited minor fluctuations which paralleled those of the intra-abdominal pressure. The 'evacuation limb' described a curve with an ascending limb continuous with the tone limb and a descending one, which descended vertically to the base line. Quantitatively, the infused CO2 volume and intrasigmoid and intra-abdominal pressures could be determined from the curve at each of the distension and expulsion sensation. Consequently, the SC compliance could be calculated. A normal sigmoidometrogram seems to reflect the normality of the sigmoid wall structure, afferent axons and central sensory pathways. Alteration of any of the components of this pathway is expected to change the measurements of SC volumes, pressure, compliance and sensation; it would also disturb the morphometry of the curve. PMID- 10817430 TI - Diverticulitis of the caecum and ascending colon: an unavoidable diagnostic pitfall? AB - PURPOSE: The infrequency of right-sided colonic diverticulitis prompted this presentation of our experiences, with emphasis on the diagnostic aspects. PATIENTS AND METHODS: Charts and documentation regarding 20 patients who underwent surgery for diverticulitis of the caecum and/or ascending colon over 22 years were reviewed. RESULTS: Eleven patients underwent pre-operative instrumental examinations: right-sided diverticulitis was recognized in five patients (two by barium enema, two by both ultrasonography and computerized tomography, one by all three examinations) and was suspected in another four. All diagnoses on merely clinical grounds--acute appendicitis in 10 patients and perforated peptic ulcer in one--were erroneous. Surgery consisted of 13 right standard or limited hemicolectomies, six conservative procedures and one Mickulicz' operation and subsequent right hemicolectomy. No operative deaths or long-term failures were reported. CONCLUSIONS: In the presence of clinical features atypical of acute appendicitis, right-sided colonic diverticulitis should be taken into account; pre-operative instrumental examinations might increase diagnostic accuracy, thereby leading to a more correct therapeutic approach. PMID- 10817431 TI - Hypoganglionic megacolon associated Chiliaditi's syndrome treated by subtotal colectomy: a case report. AB - In 1910, Chiliaditi described hepatodiaphragmatic interposition of the colon (Chiliaditi's syndrome). The condition is easily recognised on an antero posterial chest radiograph. The interposed segment of bowel is usually the hepatic flexure of the colon. The degree of interposition varies from simple wedging to complete occupation of the right subdiaphragmatic space. The management is usually conservative and surgical intervention is rarely indicated. We present a patient with hepatodiaphragmatic interposition of the colon, associated with megacolon, requiring subtotal colectomy and ileosigmoid anastomosis for severe chronic constipation. PMID- 10817432 TI - Acute colonic intramural hematoma due to blunt abdominal trauma. AB - Trauma to the colon is uncommon and accounts for only 3-5% of all blunt abdominal injuries. Among them, intramural hematoma of the colon is a rare complication and the acute form is rarer than the chronic form. We report a 37-year-old man who presented with abdominal pain followed by intestinal obstruction due to a blunt trauma. The initial diagnosis was done by sonography and proved by computed tomography (CT). Abdominal sonography also detected an increment in the size of the hematoma with progressive abdominal cramping pain that prompted urgent laparotomy. Ileocolic segmental resection with end-to-end ileocolostomy was performed and the patient recovered uneventfully. Based on our experience with a patient suffering from an intramural colonic hematoma following blunt abdominal trauma (BAT) and based on a review of the literature, we discuss the different clinical manifestations, difficulties of diagnosis, and different treatment modalities of this disease entity. We conclude that acute colonic hematoma can be diagnosed by sonography and/or CT in contrast to the early reported cases, in the pre-CT era, when they could only be diagnosed at laparotomy. Endoscopy may also be helpful for diagnosis in some cases. Although expectant therapy may be successful in some cases, the majority of the cases may need operation. PMID- 10817433 TI - Isolated blunt trauma injury to the hepatic duct. AB - Isolated bile duct injuries are quite rare. The diagnosis may be difficult. The presence of continued abdominal pain, nausea, vomiting, distention and jaundice in addition to abnormal liver function tests and often a leukocytosis mandate that a biliary tract injury be ruled out. The most important diagnostic study for confirming a biliary tract injury is paracentesis. The ERCP is then done to delineate the anatomy of the injury. Transhepatic cholangiography is an alternative to the ERCP. Treatment for a biliary duct injury must be selected on an individual basis. PMID- 10817434 TI - Biliary leakage following T-tube removal. AB - This study evaluates the incidence of biliary leakage following T-tube removal from the common bile duct (CBD) in 97 patients who underwent open CBD exploration. In 93 patients, this was following exploration for CBD stones, in two patient it was for obstructive jaundice due to hydatid disease and in a further two patients it was following CBD injury. T-tube cholangiography (TTC) was carried out 7-10 days postoperatively and, if the examination was normal, the T-tube was removed 12-14 days postoperatively (2 months for the CBD injury patients). Following T-tube removal, six patients developed severe abdominal pain, sweating and tachycardia. They were treated with antibiotics, parenteral fluids, and analgesia. Three patients settled with this management. Two patients developed sub-hepatic collections and required open drainage. One patient developed a small pelvic collection, which was aspirated transvaginally. A seventh patient was re-admitted 2 weeks following T-tube removal and laparotomy revealed biliary peritonitis. The patient died the following day. Biliary leakage following removal of a T-tube is not uncommon. It has a significant morbidity and mortality. Our experience and that of the reviewed literature suggests that the aetiology is multifactorial. The management and outcome of this complication is discussed. PMID- 10817435 TI - Carcinoid of the ampulla of Vater: presentation of a new case. AB - The appendix is the most common location for carcinoid tumors within the gastrointestinal tract. Carcinoid tumors of the ampulla of Vater is an extremely rare entity; only 71 cases of carcinoid of the ampulla of Vater have been reported in the literature to date. The clinical picture is non-specific, and endoscopy is the main diagnostic procedure; but large and deep biopsies are often needed to verify the histological nature of the tumor. Surgery is the treatment of choice. We report a new case of this rare tumor, and a review of the literature concerning the clinical findings, diagnosis, therapeutic challenge and results. PMID- 10817436 TI - A case of hemorrhagic cyst of the pancreas resembling the cystic endometriosis. AB - A 47-year-old Japanese woman with a history of epigastric pain and a recent episode of acute pancreatitis (back pain, nausea, and vomiting) and anemia was found to have a pancreatic cyst of the tail on CT-scan and ultrasonography. Especially, ultrasonography revealed the papillary solid lesion in the cyst. With the tentative diagnosis of a cystic neoplasm, distal pancreatectomy was performed. Histological examination of sections showed massive hemorrhage, surrounded fibrous connective tissue, and numerous macrophages with hemosiderin deposits; these histological findings resembled cystic endometriosis. The clinicopathological features and pathogenesis of the pancreatic endometrial cyst are discussed. PMID- 10817437 TI - Long-term and short-term survivors after pancreatectomy for pancreatic cancer. AB - Out of 63 Japanese patients with pancreatic carcinoma who underwent surgical resection, 8 short-term survivors who died within 3 months after resection and 6 long-term survivors who were alive for more than 3 years after resection were compared regarding 26 clinicopathological parameters. The 8 short-term survivors were significantly older than the 6 long-term survivors (63.7 versus 47.8 years, P = 0.0099). The mean peripheral lymphocyte count was significantly smaller in the short-term survivors than in the long-term survivors (1,212 versus 2,115 /microl, P = 0.0459). Operative blood loss was significantly larger in the short term survivors than in the long-term survivors (2,393 versus 1,043 g, P = 0.0157). The surgical margin was affected by malignant cells in 7 of the 8 short term survivors, but in only 2 of the 6 long-term survivors (P = 0.0362). Of the 8 short-term survivors, 5 were in comprehensive stage IV and 3 in stage III, while 3 of the 6 long-term survivors were in stage III, two in stage II, and one in stage I (P = 0.0487). All the 8 short-term survivors were of the comprehensive curability C, while 3 of the 6 long-term survivors were of A, one B and the other two C (P = 0.0239). Multiple regression analysis of these 6 profound factors showed that the peripheral lymphocyte count was an independent significant parameter to differentiate the short-term and long-term survivors. These findings suggest that, although the aggressive nature of pancreatic cancer has been accepted, the clinical course after pancreatectomy would also depend upon the immunological state of the patient. PMID- 10817438 TI - Fournier's gangrene in Nigeria: a review of 21 consecutive patients. AB - A review of 21 consecutive cases of Fournier's gangrene seen at the University of Port Harcourt Teaching Hospital (UPTH) in the period 1994-1998 is presented. All the patients were males with an average age of 43.4 years. Most of the patients had a predisposing factor. There was a low incidence of pre-existing medical conditions. The commonest organisms isolated were of the enterobacteria group. All the patients were treated with antibiotics, surgical debridement and frequent wound dressings with hypertonic saline, hydrogen peroxide and Eusol. In 71.4% of the cases, the wounds healed by secondary intention. The mortality was 9.5%. The deaths occurred in the older age group. We recommend surgical debridement of necrotic tissues as they present and suggest that the surgical procedures be minimal as scrotal wounds heal satisfactorily. PMID- 10817439 TI - Perceived quality of care of inguinal hernia repair: assessment before and after the procedure. AB - OBJECTIVE: Assessment of the perceived quality of care for inguinal hernia repair procedures. DESIGN: A two-step descriptive study using specific questionnaires. SETTING: A tertiary care University Hospital. SUBJECTS: Random sample of patients diagnosed of inguinal hernia in 1996. Group A, patients seen at the outpatient clinic before admission; group B, patients seen for follow-up after the surgical procedure. INTERVENTIONS: Two different questionnaires were used. MAIN OUTCOME MEASURES: validity of the questionnaire was evaluated using Cronbach's alpha value. Scores were measured using the z value. RESULTS: Total number of patients was 194. Both groups were comparable. The questionnaire was able to explain 66.3% of the variance. The Cronbach's alpha value was 0.90. Scores recorded were significantly different (P < 0.01) regarding the health care workers' attitude and hospital commodities' variables (rated as better than expected). Differences in scores for information supplied, overall results of the surgical procedure and food variables were not statistically significant. CONCLUSIONS: (i) The study of several fields where the patient's expectations are higher or lower contributes to prioritizing efforts to improve quality in the Surgery Department. (ii) The differences seen strongly suggest the need for patient's satisfaction surveys to be specific by diagnosis/dimension and adapted to patients' expectations. PMID- 10817440 TI - Comparison the efficacy of laparosonic coagulating shears and electrosurgery in laparoscopically assisted vaginal hysterectomy: preliminary results. AB - OBJECTIVE: To compare the safety and effectiveness of laparosonic coagulating shears (LCS) and electrosurgery for use in laparoscopically assisted vaginal hysterectomy (LAVH). STUDY DESIGN: In this prospective study, patients undergoing LAVH performed by one of the authors from October 1997 to January 1998 were assigned at random to the electrosurgery group (n = 20) or the LCS group (n = 20). Procedures performed with LCS or electrosurgery included coagulation and separation of infundibulopelvic or utero-ovarian round ligaments, vesico-uterine visceral peritoneal fold dissection, and anterior and posterior colpotomy. Outcome measures were operative time, blood loss, decrease in hemoglobin values, and length of hospitalization. RESULTS: The mean operative time (90+/-22.9 min versus 80.3+/-17.1 min, P = 0.391), blood loss (308+/-167 ml versus 250+/-104 ml, P = 0.11), and hemoglobin decrease (1.57+/-0.769 mg/dl versus 1.36+/-0.886 mg/dl, P = 0.55) were slightly greater in the LCS group than in the electrosurgery group, although these differences were not statistically significant. The length of hospital stay was similar in the two groups (5 days). No patients developed serious complications related to electrosurgery or LCS. CONCLUSION: Our findings indicate that LCS is as safe and effective as electrosurgery, and may offer an alternative option for patients undergoing LAVH. PMID- 10817441 TI - Molecular genetics of migraine headaches: a review. AB - Following the recent discovery of neural calcium channel mutations in familial hemiplegic migraine, genetic linkage and association studies have been performed world-wide in an effort to unveil the genetic basis of the more common types of migraine too. Mutations in neural calcium channels, mitochondrial DNA, serotonin receptors and transporter, dopamine receptors and genetic prothrombotic risk factors have been especially investigated and are discussed here. No unambiguous conclusions have, however, been reached. FHM remains an isolated success story in the quest for the genetic basis of migraine. PMID- 10817442 TI - Migraine patients show an increased density of dopamine D3 and D4 receptors on lymphocytes. AB - Recent studies have revealed peculiar functional and genetic features of dopamine receptors in migraine. As peripheral blood lymphocytes (PBL) may represent a tool for peripheral detection of neuroreceptors, we compared the expression of dopamine D3 (DRD3) and D4 (DRD4) receptors on PBL in migraine patients and in healthy controls using radioligand binding assay techniques in the presence of antidopamine D2-like receptor antibodies. The dopamine D2-like receptor agonist [3H]7-OH-DPAT was used as a radioligand. An increased density of both DRD3 (P=0.0006) and DRD4 (P=0.002) on PBL was observed in migraineurs compared with controls. This up-regulation might reflect central and/or peripheral dopamine receptor hypersensitivity due to hypofunction of the dopaminergic system. These findings support the view that dopamine D2-like receptors are involved in the determination of the so-called migraine trait, which may help to elucidate several clinical features of the disease. PMID- 10817443 TI - Are the periodic changes of neurophysiological parameters during the pain-free interval in migraine related to abnormal orienting activity? AB - OBJECTIVE AND METHODS: Migraine patients are characterized by increased amplitude and reduced habituation of contingent negative variation (CNV). Furthermore, the CNV underlies periodic changes during the pain-free interval, being maximal before attack. The periodicity of CNV is related to periodic changes in habituation, probably due to variation of orienting activity during the pain-free interval. CNV and orienting response (OR) were studied in 20 females suffering from migraine without aura and in 12 matched healthy females. The neurophysiological recordings in the group of patients were performed 1-4 days before and 4 days after a migraine attack. The amplitudes and habituation of early and late components and total CNV were calculated. The OR was assessed using the habituation of the skin conductance response (SCR) and alpha blocking (AB). The non-parametric tests were employed for statistical analysis. RESULTS: There were no differences between the two groups for habituation of all CNV components and of SCR following an attack. However, the habituation of AB was significantly reduced in migraine. Before attack we observed a significantly reduced habituation of the early and total CNV and of the AB compared to controls and recordings performed after an attack. The habituation of the late component and of SCR remained unchanged. CONCLUSIONS: The abnormal habituation could be explained by the periodic changes of physiological parameters during the pain free interval. The impaired habituation of early CNV in migraine is associated with increased orienting activity seen only in the central component (AB) of OR. PMID- 10817444 TI - Comparison of the efficacy of zolmitriptan and sumatriptan: issues in migraine trial design. AB - In this international, multicentre, double-blind, placebo-controlled, single attack study, 'triptan naive' migraine patients were randomized in an 8:8:1 ratio to receive zolmitriptan 5 mg, sumatriptan 100 mg or placebo. The all-treated analysis included 1058 patients who took study medication. The primary endpoint, complete headache response, was reported by 39%, 38% and 32% of patients treated with zolmitriptan, sumatriptan and placebo, respectively, with no significant difference between treatment groups. In patients with moderate headache at baseline, complete response was significantly greater following zolmitriptan than after placebo (48% vs. 27%; P=0.01); there was no significant difference between sumatriptan and placebo groups (40% vs. 27%). In patients with severe baseline headache (where a greater reduction in headache intensity is required for a headache response), there was no significant difference between any groups in complete headache response rates. For secondary endpoints, active treatment groups were significantly superior to placebo for: 1-, 2- and 4-h headache response (e.g. 2-h headache response rates: zolmitriptan 59%; sumatriptan 61%; placebo 44%; P < 0.01 vs. placebo); pain-free response rates at 2 and 4 h; alleviation of nausea and vomiting; use of escape medication and restoration of normal activity. The incidence of adverse events was similar between zolmitriptan and sumatriptan groups but was slightly lower in the placebo group. The lack of difference between active treatments and placebo for complete response probably reflects the high placebo response obtained, which is probably a result of deficiencies in trial design. For example, the randomization ratio may result in high expectation of active treatment. Thus, while ethically patient exposure to placebo should be minimized, this must be balanced against the scientific rationale underpinning study design. PMID- 10817446 TI - Motor cortex excitability in patients with migraine with aura and hemiplegic migraine. AB - We studied the excitability of the motor cortex using transcranial magnetic stimulation (TMS) in 12 patients with migraine with aura (MA) and nine patients with familial hemiplegic migraine (FHM). Motor thresholds at rest, the duration of the cortical and peripheral silent period and intracortical inhibition and facilitation using paired-pulse TMS at intervals of 2 to 15 ms were measured with patients free of attacks for at least 48 h. In contrast to previous reports we could not find any significant differences between patient groups and compared to controls (n=17) in the parameters tested. The results suggest that there are no interictal changes of excitability of the motor cortex in migraine. This study does not support the concept of general cortical hyperexcitability in migraine secondary to a genetic predisposition or a structural alteration of inhibitory interneurones in the cortex due to repeated parenchymal insults during attacks. PMID- 10817445 TI - Effects of the anti-migraine drug sumatriptan on muscle energy metabolism: relationship to side-effects. AB - Sumatriptan succinate (Imitrex) is a 5-HT (5-hydroxytryptamine) agonist used for relief of migraine symptoms. Some individuals experience short-lived side effects, including heaviness of the limbs, chest heaviness and muscle aches and pains. The effects of this drug on skeletal muscle energy metabolism were studied during short submaximal isometric exercises. We studied ATP flux from anaerobic glycolysis (An Gly), the creatine kinase reaction (CK) and oxidative phosphorylation (Ox Phos) using 31P nuclear magnetic resonance spectroscopy (31P MRS) kinetic data collected during exercise. It was found that side-effects induced acutely by injection of 6 mg sumatriptan succinate s.c. were associated with reduced oxygen storage in peripheral skeletal muscle 5-20 min after injection as demonstrated by a transient reduction in mitochondrial function at end-exercise. These results suggest that mild vasoconstriction in peripheral skeletal muscle is associated with the action of sumatriptan and is likely to be the source of the side-effects experienced by some users. Migraine with aura patients were more susceptible to this effect than migraine without aura patients. PMID- 10817447 TI - Cluster of MMPI personality profiles in chronic tension-type headache and predictable response to Fluoxetine. AB - A study was made of the personality profiles of a sample of 51 patients with chronic tension-type headache (CTH) employing the Minnesota Multiphasic Personality Inventory (MMPI). Two clusters were obtained by applying multivariate classification techniques: cluster 1 (with elevations on scales Hy, D and Hs only) and cluster 2 (exhibiting elevations on most scales except Pd, Mf and Si). Fifty subjects without chronic pain or known psychiatric disorders, and extracted from the same social setting as the patients, completed the MMPI as a control group. Fluoxetine treatment was started in the CTH group, with follow-up over a 1 year period. Chi-squared analysis correlating the clusters obtained to different pain-related variables and epidemiological parameters revealed a significant association to sex only. There were no differences in therapeutic response between the two clusters. However, the patients belonging to the less perturbed cluster who exhibited profiles analogous to those of the control population showed significant improvement with respect to the global sample and their own cluster. PMID- 10817448 TI - Orientation discrimination thresholds in migraine: a measure of visual cortical inhibition. AB - Orientation discrimination is a visual task dependent on inhibitory mechanisms in the visual cortex. In this study, orientation discrimination thresholds for bar and grating patterns were measured at two visual field locations in subjects with migraine with (n = 20) and without aura (n = 20) and in migraine-free control subjects (n = 20). No statistically significant differences were found between migraine groups and the control group on either task at foveal or peripheral visual field locations. No significant correlations were found between psychophysical thresholds and age, total lifetime auras or total lifetime migraine episodes. However, a trend was seen toward slightly impaired performance on the two foveal tasks in a subgroup of subjects with the highest total lifetime aura count. Thus we have found no convincing evidence that impaired cortical inhibitory mechanisms are a predisposing characteristic in migraine, but cannot rule out the possibility that cortical inhibitory mechanisms may be adversely affected by repeated visual auras. PMID- 10817449 TI - A SUNCT case with response to surgical treatment. PMID- 10817450 TI - Animal definition: a necessity for the validity of animal experiments? AB - In most scientific journals, experimental animals are described poorly. Whether this is scientifically justified is discussed in this article. It was concluded that when laboratory animals are used in scientific experiments, which almost always are of a quantitative nature, a detailed animal definition is imperative. PMID- 10817451 TI - Guidelines for reporting the results of experiments on fish. AB - A detailed account of experimental design, including an accurate description of the animals used, is an essential part of good research practice. Without these details, the reader will be unable not only to form an opinion on the significance of the findings but also to repeat the experiment in another laboratory. This paper presents suggested guidelines for reporting experimental studies using fish. PMID- 10817452 TI - R-weighting provides better estimation for rat hearing sensitivity. AB - Since sounds may induce physiological and behavioural changes in animals, it is necessary to assess and define the acoustic environment in laboratory animal facilities. Sound studies usually express sound levels as unweighted linear sound pressure levels. However, because a linear scale does not take account of hearing sensitivity-which may differ widely both between and within species at various frequencies-the results may be spurious. In this study a novel sound pressure level weighting for rats, R-weighting, was calculated according to a rat's hearing sensitivity. The sound level of a white noise signal was assessed using R weighting, with H-weighting tailored for humans, A-weighting and linear sound pressure level combined with the response curves of two different loudspeakers. The sound signal resulted in different sound levels depending on the weighting and the type of loudspeaker. With a tweeter speaker reproducing sounds at high frequencies audible to a rat, R- and A-weightings gave similar results, but the H weighted sound levels were lower. With a middle-range loudspeaker, unable to reproduce high frequencies, R-weighted sound showed the lowest sound levels. In conclusion, without a correct weighting system and proper equipment, the final sound level of an exposure stimulus can differ by several decibels from that intended. To achieve reliable and comparable results, standardization of sound experiments and assessment of the environment in animal facilities is a necessity. Hence, the use of appropriate species-specific sound pressure level weighting is essential. R-weighting for rats in sound studies is recommended. PMID- 10817453 TI - Coronary artery anomalies and aortic valve morphology in the Syrian hamster. AB - In the Syrian hamster, anomalies in the origin of the left coronary artery are significantly associated with the bicuspid condition of the aortic valve. In this species, bicuspid aortic valves are expressions of a trait, the variation of which takes the form of a phenotypic continuum, ranging from a tricuspid aortic valve with no commissural fusion to a bicuspid aortic valve with the aortic sinuses located in ventrodorsal orientation and devoid of any raphe. The intermediate stages of the continuum are represented by tricuspid aortic valves with a more or less extensive fusion of the ventral commissure and bicuspid aortic valves with a more or less developed raphe located in the ventral aortic sinus. The present study was designed to decide whether there is a gap between tricuspid and bicuspid aortic valves regarding the incidence of coronary artery anomalies, or whether this incidence varies according to the different tricuspid and bicuspid morphotypes of the continuum. The study was carried out in Syrian hamsters belonging to a single inbred family with a high incidence of tricuspid aortic valves with fusion of the ventral commissure, bicuspid aortic valves, and anomalies in the origin of the left coronary artery, i.e. single right coronary artery ostium in aorta, anomalous origin of the left coronary artery from the pulmonary artery, and anomalous origin of the left coronary artery from the dorsal aortic sinus. The specimens were examined by means of a stereomicroscope and, in several cases, scanning electron microscopy was also used. The relationships between anomalous coronary artery patterns and aortic valve morphologies were tested using a logistic regression model. The results obtained indicate that there is no discontinuity between tricuspid and bicuspid aortic valves regarding the incidence of coronary artery anomalies. The probability of occurrence of anomalous coronary artery patterns increases continuously according to the deviation degree of the aortic valve from its normal (tricuspid) design. The present findings suggest that in the Syrian hamster, the morphogenetic mechanisms involved in the formation of congenital anomalous aortic valves and anomalies in the origin of the left coronary artery, respectively, are strongly related from an aetiological viewpoint. PMID- 10817454 TI - Dietary fats influence consumption and metabolic measures in male and female laboratory mice. AB - Juvenile and adult male and female Swiss mice in metabolism cages were fed one of four specially-formulated, pelleted diets containing respectively 8% saturated vegetable fat, 8% soya oil, 8% olive oil and 2% soya oil. The identities of the diets were hidden from the experimenter. Subjects were individually housed in metabolism cages and their consumption of food, growth and eliminative activities were measured. Clearly, these non-isocaloric diets differed in palatability, producing complex effects on growth as well as metabolic measures. Many indices were influenced by age, sex, and the duration of dietary exposure. Interactions between factors were common. Dietary fats appear to have subtle effects on the physiology and behaviour of rodents and may account for some differences between studies. PMID- 10817455 TI - A new technique to determine hydrogen excreted by gnotobiotic rats. AB - A new system, that allowed the monitoring of hydrogen (H2) excretion by gnotobiotic rats without affecting their defined microbial status, was developed. The system consists of an isolator containing a chamber for an experimental animal, and a life-support system (LSS), with a sampling port outside the isolator connected to it. H2 accumulation in the system was measured by analysing a defined volume of gas after removal. H2 concentrations were determined with an electrochemical cell or by gas chromatography. To validate this technique, H2 excretion by germ-free (GF) and mono-associated rats fed a chemically defined diet was measured after oral application of lactulose. Mono-associated rats had been obtained by colonizing GF rats with a H2-producing Clostridium perfringens type A strain isolated from human faeces of a healthy volunteer. Application of 50 mg lactulose to the mono-associated rats resulted in a significant increase in H2 excretion. The net H2 excretion was 7.82+/-1.28 ml H2 in 12 h corresponding to a net maximal rate of 1.1+/-0.3 ml H2/h. In contrast, in experiments with GF rats, less than 0.13 ml H2 were detectable within 12 h. The technique presented is a useful tool for studying bacterial H2 metabolism in vivo under gnotobiotic conditions. PMID- 10817456 TI - Salivary and plasma cortisol response to adrenocorticotropin administration in pigs. AB - Two experiments were conducted to determine the responsiveness of salivary and plasma cortisol to acute (i.v.), depot (i.m.) and chronic (repeated i.m.) adrenocorticotropin (ACTH) administration in swine. In Experiment 1, barrows (castrated pigs) were assigned to one of three injection treatments: (1) saline i.m. (SHAM1, n=2); (2) 0.75 IU/kg BW ACTH in saline i.v. (ACUTE, n=2); (3) 2.25 IU/kg BW ACTH in gel i.m. (DEPOT, n=3). Total cortisol concentrations were determined for concurrent saliva and blood samples. Correlations between salivary and plasma cortisol within treatments were: SHAM1, r=0.60; ACUTE, r=0.58; DEPOT, r=0.79. In Experiment 2, barrows were assigned to one of two injection treatments: (1) gel i.m. (SHAM2, n=3); (2) 2.25 IU/kg BW ACTH in gel i.m. (CHRONIC, n=4). The injections occurred every 6 h for a total of eight injections. Concurrent saliva and blood samples were obtained every 3 h for 72 h followed by an increasing sampling interval until day 6. Overall correlations between salivary and plasma cortisol were: SHAM2, r=0.30 and CHRONIC, r=0.61. Experiment 1 found that the relationship between salivary and plasma cortisol was stronger during longer (DEPOT) than shorter (ACUTE) ACTH stimulation. Experiment 2 found a strong relationship between the two measurements during chronic ACTH stimulation, but that relationship weakened after ACTH stimulation ceased. PMID- 10817457 TI - A simplified technique for non-surgical catheterization of the vena cava cranialis in pigs and an evaluation of the method. AB - Restraint and venepuncture affect blood parameters and are stressful for pigs. Therefore, when experiments require frequent blood sampling, catheterization is the best option, both ethically and scientifically. A simplified non-surgical method for catheterization of the vena cava cranialis in sows is described. The vena cava is catheterized via the vena jugularis externa, which is punctured while the sow is restrained by nose snaring. The procedure can be carried out by two experimenters in 5 min, it requires little equipment and can be performed under farm or very simple experimental conditions. Results from 89 catheterizations are reported. Seventy-three of the catheters remained functional for 5 days or more. There was a tendency for a higher success rate in dry sows than in highly pregnant sows. The only behavioural effect observed after catheterization was a short-lasting increase in motivation for wallowing in some sows. No illness that could be attributed to the catheterization occurred. PMID- 10817458 TI - Fighting in NIH/S male mice: consequences for behaviour in resident-intruder tests and physiological parameters. AB - Fighting is known to occur frequently in male mouse groups. In this study with outbred NIH/S mice, the possible impact of individual aggressiveness on fighting in groups and on the social status of animals was studied. Male mice were pre tested in a resident-intruder (RI) test and rated as initially aggressive or non aggressive according to their attack behaviour against an intruder. Thereafter they were randomly allocated to new social groups, with four mice per cage. Fighting in groups was increased when several initially aggressive animals were included in the group. Within the groups, animals were rated as dominants and subordinates according to their behaviour toward a strange intruder introduced into their home-cage (Group Intruder, GI) test and the occurrence of wounds. Additionally, subordinates were divided into aggressive and non-aggressive categories according to their behaviour in the second RI test, which was performed 3 weeks after grouping. The behaviour in the RI test prior to group housing did not predict the individual social status or possibility of being wounded in the new social environment. On the other hand, the social relationships in the new group affected the behaviour in a subsequent RI test. All dominants showed aggressive behaviour during the second RI test. Those subordinates which behaved aggressively during this test received the most numerous and serious wounds, suggesting that in the new groups their interactions with the other group members were mostly aggressive. The reduced weight of epididymal adipose tissue in dominant and aggressive subordinates may indicate that they had fought continuously. Social status or levels of fighting in a group did not affect individual weight gain or the other physiological parameters measured. The wounded animals had enlarged spleens and reduced weights of epididymal adipose tissue, which were probably the results of increased activity of the immune system and reduced welfare, respectively. In conclusion, individual aggressiveness seems to be greatly affected by the demands of the social environment. Fighting in mouse groups leading to wounded animals may have effects on physiological research parameters. PMID- 10817459 TI - Rapid oral endotracheal intubation with a fibre-optic scope in rabbits: a simple and reliable technique. AB - The orotracheal intubation of rabbits is complicated by their oropharyngeal anatomy. Numerous techniques to intubate rabbits have been described; however, these methods require specialized devices, tracheostomy, or are performed in a blind fashion. We describe a technique for the intubation of the rabbit under direct visualization with a standard, small-bore, fibre-optic laparoscope, which is both rapid and simple to perform. PMID- 10817460 TI - Adverse effects on growth rates in rats caused by buprenorphine administration. AB - As a routine postoperative treatment, a single dose of buprenorphine was given to rats at a dose of 0.05 mg/kg subcutaneously. However, some rats developed abnormal secretions around the nose and mouth and some animals died 3-5 days after surgery and analgesic treatment. At autopsy a yellow fibrous mass was found in the stomach and intestines. Observations of animals given buprenorphine revealed an abnormal ingestion of bedding material. This caused a disturbance to normal digestion, with gastric distension, weight loss or decreased growth rate, constipation and occasionally death. In this study rats were monitored for 6 days following surgery and analgesic treatment. A comparison of growth rates was made between rats given saline and buprenorphine or nalbuphine and between animals kept on bedding or grid floors for the first 24h after treatment. Of the animals held on bedding, the buprenorphine-treated animals did not lose weight as the other animals did, but had on the other hand a decreased growth rate during the measuring period of 6 days after surgery. When denied access to bedding for the first 24 h after surgery, rats given saline or nalbuphine had a reduced weight gain over the first 24 h, similar to the groups held on bedding. Rats held on grid floors and given buprenorphine continued to gain weight for the first 24 h. From day 3, there was no significant difference between the groups, which all gained weight. PMID- 10817461 TI - Effects of repeated anaesthesia with ketamine/medetomidine and of pre-anaesthetic administration of buprenorphine in rats. AB - Two groups of rats were anaesthetized at weekly intervals for 6 weeks with either ketamine/medetomidine alone (60 mg/0.4 mg/kg i.p.) or ketamine/medetomidine (45 mg/0.3 mg/kg i.p.) one hour following buprenorphine (0.05 mg/kg s.c.). Animals that received buprenorphine had longer periods of surgical anaesthesia (P = 0.04) and a greater depression of both mean pedal withdrawal score (P < 0.01) and mean respiratory rate (P = 0.014). Mean total duration of anaesthesia was also greater in the buprenorphine group on day 1. Sleep times reduced with successive doses of anaesthetic in the buprenorphine group (P = 0.024). Two animals in the buprenorphine group died. Repeated anaesthesia with ketamine/medetomidine alone was not associated with anaesthetic mortality. These results indicate that although buprenorphine has a clear anaesthetic-sparing effect, its use with ketamine/medetomidine may be associated with an increased risk of anaesthetic related mortality. PMID- 10817462 TI - Granulomatous inflammation in the tails of mice associated with Mycobacterium chelonae infection. AB - This report describes a case series of granulomatous inflammation in the tails of seven immunocompromised mice. The disease was associated with Mycobacterium chelonae infection. The source and route of infection remained unclear. Spontaneous infection with M. chelonae has not previously been documented in mice. We conclude that M. chelonae, like in humans, should be considered as a facultative pathogen in laboratory animals, particularly under immunosuppressive conditions. PMID- 10817463 TI - Attempted eradication of salmonellosis from a colony of short-tail grey opossums (Monodelphis domestica). AB - Prior to re-housing a colony of laboratory short-tail grey opossums, the animals were found to harbour salmonella. This paper describes an unsuccessful attempt to eradicate the infection from the colony by means of antibiotic treatment and hygienic measures. A pilot treatment of five animals which received enrofloxacin 10 mg/kg for 5 days appeared to be successful in that no salmonellae were recovered from faeces or organs sampled after treatment. The process was repeated on the whole colony prior to a change of accommodation but 2 animals were found to be still infected, 5 weeks after cessation of treatment. PMID- 10817464 TI - Sexual abstinence at age 21 in New Zealand: the importance of religion. AB - Most research on adolescent sexual behaviour has focused on early initiation and consequent risks. We have instead examined the circumstances of young people who have not had sexual intercourse before age 21, in order to throw light on the ways in which young people might resist societal pressures for early sexual intercourse. The sample was a cohort born in Dunedin, New Zealand in 1972/73, formed at age 3, and followed with regular assessments of personal, family and educational functioning to age 21. At age 18 and 21 information on sexual behaviour was collected, using a computer presented questionnaire. The response rate at age 21 was 935/1020 (91.7%) of the survivors of the original cohort. Overall 11.3% of the men and 8.1% of the women reported never having sexual intercourse. Sex with a man was reported by 20 men (4.5%), of whom only two reported having sex only with men. Being first born and being persistently involved in religious activities, measured at both 11 years and 21 years, were significant predictors of abstinence for both sexes. Examination of perceptions of an ideal lifestyle, sexual behaviour and religious involvement showed that religion was an important factor in decisions to delay sexual intercourse past age 20, especially for men. It would be helpful to examine further the features of moral decision making which are characteristic of religious experiences. PMID- 10817465 TI - Contamination of medicine injection paraphernalia used by registered medical practitioners in south India: an ethnographic study. AB - While considerable attention has been directed at the important role of intravenous drug use in the spread of human immunodeficiency virus (HIV) and hepatitis B, little research to date has been conducted on the role of medicine injections in disease transmission. This is the case despite the fact that (a) the number of medicine injections is several orders of magnitude greater than injections of illegal drugs and (b) the networks of people potentially affected by contaminated medicine injection paraphernalia is far wider. In this article we examine the medicine injecting practices of a random sample of 40 registered medical practitioners (RMP) who have not had formal training in allopathic medicine (do not have MBBS or MD degrees) in Tamil Nadu, India. Attention is drawn to: (a) the lack of vigilance practitioners exercise in maintaining hygienic needles and syringes, (b) their perceptions of what constitutes acceptable hygienic procedure and (c) how patients respond in contexts where they are able to purchase disposable needles and syringes directly from practitioners or from the open market prior to visiting a practitioner. Study results are a cause for alarm and indicate widespread contamination of injection paraphernalia as well as common reuse of disposable needles. The study was confined to RMPs and the researchers strongly suggest that future studies of MBBS trained doctors practising in the public and private sectors be carried out. A structured observation instrument developed to record needle and syringe contamination during the process of injection administration is provided. PMID- 10817466 TI - Hong Kong Chinese perceptions of the experience of unrelated bone marrow donation. AB - This paper presents a qualitative perspective of the Chinese experience of unrelated bone marrow donation. A total population of 37 Chinese men and women, residing in Hong Kong who had donated bone marrow to an unrelated recipient were interviewed and asked their retrospective perceptions and experiences during the process of unrelated bone marrow donation. The majority was female (60%) and single (59.3%). The median age was 30.9 years. The main themes drawn from thematic content analysis included: association with bad and good fortune, religious concerns associated with complete bodies, barriers posed by the extended family, fulfilling personal identities linked to perceptions of altruism, reciprocity and generosity and the differences between the expectations and reality of the experience of bone marrow donation. These accounts indicate how, for Chinese populations, the act of body fluid donation cannot be isolated from the experiences of life-long immersion in the dominant social and cultural processes of the time. In addition, Confucian notions of the body, the power of body fluids to pollute, the importance of blood and bones as sources of vital energy and the process of bodily transference into the after life are highlighted. Furthermore, what is also identifiably Chinese is that donation is more of a self-fulfilling act than a social act with very little familial or social recognition being accorded in the public world. The conclusion highlights how donors entered into the decision to donate with little awareness of the long term consequences. The process, however, was seen by the donors to be beneficial. A compelling argument is made for more educational and emotional support regarding the experience of bone marrow donation from Hong Kong's formal service providers. PMID- 10817467 TI - Chronic physical illness, psychiatric disorder and disability in the workplace. AB - While agreement is growing that mental illness burdens the North American economy, how it impacts productivity--particularly compared to physical illness- is unclear. Hypothesizing that lost work days are only the tip of the iceberg, we also examined the association of mental and chronic physical illness with partial work days and days requiring extra effort to function. Data from 4225 employed individuals, aged 18-54, were analyzed. These were a subset of respondents to the Ontario Health Survey's Mental Health Supplement, a 1990/91 epidemiologic survey of households across Ontario, Canada. Psychiatric disorder was assessed using the University of Michigan' modification of WHO's Composite International Diagnostic Interview (UM-CIDI). Similar to US reports, professional/managerial groups had lower rates of affective and anxiety disorders and fewer disability days compared to the rest of the workforce. However, no single occupational group was consistently at greater risk for either physical or psychiatric problems. Even after accounting for sociodemographic characteristics and work conditions, mental and physical status had clear, but different, impacts on productivity. Physical conditions alone had a fairly constant effect across all types of disability days and were the largest contributor to total work day loss. They also significantly impacted partial and extra effort days but were far less important than conditions involving a mental disorder. Respondents with mental health problems, either alone or in combination with physical illnesses, appeared more likely to go to work but to require greater effort to function. WHO projects that mental illness will become the second most important cause of global disease burden in the next century. Our findings suggest that among working individuals, it affects productivity more subtly than does physical illness. However, with an estimated eight percent of Ontario's workforce experiencing more than two months annually of decreased productivity, it still incurs significant social and economic costs. PMID- 10817468 TI - The three-year persistence of depressive symptoms in men and women. AB - Little is known about sex differences in the course of depression. The results of the few surveys of the general population are inconsistent. Furthermore, the generalizability of most of these findings is limited. First, only a handful of studies of the general population use a prospective design to estimate the persistence of depression in women and men. Second, in some studies only particular subgroups of the general population are sampled, and finally, depression persistence is often measured using unconventional short time intervals. Using data from five waves of the Flemish subsample of the Panel Study of Belgian Households (PSBH)--a probability sample of 1168 (52.5%) women and 1055 (47.5%) men--we tried to go beyond these shortcomings. Depression persistence is estimated using a self-report inventory on four occasions separated by intervals of one year (1993, 1994, 1995, 1996). Results show a significant influence of sex, other sociodemographic characteristics, and depression severity at baseline on three-year depression persistence. Women experience more chronicity, a difference that can be partially ascribed to sex differences in employment status, education and marital status. Persistence in linked to marital status in women. In men persistence is associated with level of education and with employment status. Depression severity at baseline, however, largely explains the influence of these social conditions on depression persistence. PMID- 10817469 TI - Theoretical perspectives on suicide in gay men with AIDS. AB - Suicide among gay men with AIDS constitutes a significant theoretical and practical problem. In order to understand this problem it is necessary to consider the specific circumstances that surround these individuals. Whilst there are many theories of suicide with which to address the problem, few are able to capture the special context within which gay men with AIDS live. This article explores three broad concepts from the theoretical literature on suicide that have explanatory potential in this unique context. The three concepts are: the search for meaning in the midst of suffering, Shneidman's [Shneidman, E., 1985. Definition of Suicide. John Wiley, New York] notion of unendurable psychological pain, and hopelessness. This article attempts to demonstrate how these three concepts can incorporate the experiences of gay men with AIDS and contribute to understanding the problem of suicide in this population. PMID- 10817470 TI - God should give daughters to rich families only: attitudes towards childbearing among low-income women in Punjab, Pakistan. AB - We evaluated perceptions and experiences of bearing sons and daughters among 42 women in Punjab, Pakistan, with special emphasis on son preference, changes in women's status within the marital family and resulting health effects. Data were collected through repeated, in-depth interviews in Urdu or Punjabi in an urban area in Lahore and a village 40 km. outside of Lahore. For triangulation purposes, four focus group discussions were performed with additional women, as well as in-depth interviews with eight mothers-in-law, three traditional practitioners and three medical practitioners. In general, these women felt that they had limited control over their lives, and this was exemplified by early marriages, high expectations on newly wed women to conceive and poor access to contraceptives. Women frequently expressed a strong preference for sons, mostly for economic reasons, reflecting women's subordinate position in society and the low economic value placed on women's work. Mothers of sons mainly discussed health problems during pregnancy and health effects of repeated childbearing. Mothers of daughters and women without children spoke of harassment in the family as well as in society. The results should be of importance in the public health planning in Pakistan as well as for those engaged in women's health issues internationally. PMID- 10817471 TI - Clinical risk and collective competence in the hospital emergency department in the UK. AB - The hospital emergency department (ED) is a risky environment, often subject to litigation for negligence. Risk is defined as an avoidable increase in the probability of an adverse outcome for a patient. With the aim of identifying the sources of risk, this study carried out participant observation and collected critical incidents in two EDs in the UK for a period of 30 months. Active failures included delay in beginning investigations or treatment, failure to obtain diagnostic information, misinterpretation of diagnostic information and the administration of inappropriate treatment. Three latent conditions underlay these failures: patients' unrestricted access to the ED, cognitive errors by individual members of staff and a strict horizontal and vertical division of labour. An analysis of the incidents resulting from the third latent condition identified a contradiction between the division of labour and working conditions in the ED. The paradigm circumstances under which this contradiction can result in active failures are described. The management of risks arising in this way could be improved by developing a workplace culture in which 'sapiential authority'--authority derived from experience, special access to information or being at hand in an emergency--is recognised in addition to authority derived from a formal status. However, as long the contradictions between the division of labour and working conditions remain, accidents should be considered normal events. PMID- 10817472 TI - Ambiguity: exploring the complexity of roles and boundaries when working with volunteers in well woman clinics. AB - Recently in the United Kingdom there has been a focus on strengthening the participation of volunteers in the National Health Service (NHS), particularly within primary health care services. Despite this focus, the challenges and opportunities of collaborative working between volunteer and paid workers has been under explored. Drawing on the findings from an ethnographic study, the concept of ambiguity is used to illuminate the complexity of roles and boundaries when working with volunteers in community well woman clinics. The findings are based on data collected during an eighteen month period of participant observation, interviews with 26 volunteer and paid workers and a review of the clinics' operational policy documents. Recognising the ambiguous position of volunteers in formal organisations is central to gaining an insight into their experiences of volunteering and into their relationships with paid workers. Volunteers within formal organisations are likely to experience role ambiguity, as often roles and expectations may not be clearly differentiated. Additionally, volunteering within formal organisations is inherently contradictory, as it is both work and a leisure pursuit. A lack of understanding as to which element, either work or leisure, is being emphasised can lead to tensions in relationships between volunteer and paid workers. Learning to work with ambiguity is central to being able to work effectively with volunteers and strategies for managing ambiguity are proposed. PMID- 10817473 TI - Women's experiences of maternity care: satisfaction or passivity? AB - This study is a first attempt to describe Lebanese women's responses to the medical management of their pregnancy and delivery. A qualitative approach in data collection and analysis was adopted to gain an in-depth view of women's perceptions. Women of any parity undergoing a normal vaginal delivery during the three months preceding the interview were interviewed in different areas in Lebanon: one urban, one semi-rural and two remote rural. Childbirth for all the women interviewed was managed within the medical system. Findings show that women accord total trust to their physicians, and very rarely question the usefulness of many routinely applied procedures, even those which the literature shows are unnecessary. When probed, women report that many aspects of the technical care are intimidating and that they experience discomfort with these procedures. Women are more vocal about patient-provider communication and value good interaction with their provider. The extent of passivity and feelings of discontent women have varies according to their social class and the amount of psychosocial support they receive throughout the process of childbirth. PMID- 10817474 TI - The social gradient in life expectancy: the contrary case of Okinawa in Japan. AB - This paper examines the social gradient theory of health and life expectancy presented by Evans and his colleagues [Evans, R.G., Barer, M.L. and Marmor, T.R. (Eds.), 1994. Why are some People Healthy and others not? The Determinants of Health of Populations. Aldine de Gruyter, New York]. They maintain that social hierarchy is the determining factor in the health of large populations largely because it promotes differences in stress or the ability to cope with stress. For example, as Japan has risen to the top ranks of the economic hierarchy of nations in the late 20th century, Japanese life expectancy improved dramatically. Evans [Evans, R.G., 1994. Introduction. In: Evans, R., Barer, M., Marmor T. (Eds.), Why are some People Healthy and others not? The Determinants of Health of Populations. Aldine de Gruyter, New York, pp. 3-26.] notes that something lies behind this rapid increase in longevity and the major change was the hierarchical position of Japan relative to the rest of the world. However, we reviewed life expectancy data within Japan and found that Okinawans traditionally rank at the top in health and life expectancy and at the bottom in socioeconomic indicators. We find that the social gradient thesis does not apply in Japan and suggest that what is more important for health are health lifestyles, especially diet and social support. More research is needed to assess the validity of the social gradient thesis if it is to be used on a cross-national basis. PMID- 10817475 TI - Socio-economic status and the utilisation of physicians' services: results from the Canadian National Population Health Survey. AB - This paper assesses the extent to which Canada's universal health care system has eliminated socio-economic barriers in the use of physician services by examining the role of socio-economic status in the differential use of specific, publicly insured, primary and specialist care services. Data from the 1994 National Population Health Survey, a nationally representative survey, were analysed using multiple logistic regression. In order to control for the association between primary and specialist utilisation, a two-staged least squares method was used for models explaining specialist utilisation. Health need, as measured by perceived health status and number of health problems, was found to be consistently associated with increased physician utilisation, for both primary and specialist visits. Whereas the likelihood of an individual making at least one visit to a primary care physician was found to be independent of income, those with lower incomes were more likely to be frequent users of primary care, that is, make at least six visits to a primary care physician. Even after adjusting for the greater utilisation of primary care services by those in lower socio-economic groups, and, therefore, their higher exposure to the risk of referral, the utilisation of specialist visits was greater for those in higher socio-economic groups. Canadians lacking a regular medical doctor were less likely to receive primary and specialist care, even after adjustments for socio economic variables such as income and education. Although financial barriers may not directly impede access to health care services in Canada, differential use of physician services with respect to socio-economic status persists. After adjusting for differences in health need, Canadians with lower incomes and fewer years of schooling visit specialists at a lower rate than those with moderate or high incomes and higher levels of education attained despite the existence of universal health care. PMID- 10817476 TI - Income inequality, social cohesion and the health status of populations: the role of neo-liberalism. AB - There has been a recent upsurge of interest in the relationship between income inequality and health within nations and between nations. On the latter topic Wilkinson and others believe that, in the advanced capitalist countries, higher income inequality leads to lowered social cohesion which in turn produces poorer health status. I argue that, despite a by-now voluminous literature, not enough attention has been paid to the social context of income inequality--health relationships or to the causes of income inequality itself. In this paper I contend that there is a particular affinity between neo-liberal (market-oriented) political doctrines, income inequality and lowered social cohesion. Neo liberalism, it is argued, produces both higher income inequality and lowered social cohesion. Part of the negative effect of neo-liberalism on health status is due to its undermining of the welfare state. The welfare state may have direct effects on health as well as being one of the underlying structural causes of social cohesion. The rise of neo-liberalism and the decline of the welfare state are themselves tied to globalization and the changing class structures of the advanced capitalist societies. More attention should be paid to understanding the causes of income inequalities and not just to its effects because income inequalities are neither necessary nor inevitable. Moreover, understanding the contextual causes of inequality may also influence our notion of the causal pathways involved in inequality-health status relationships (and vice versa). PMID- 10817477 TI - Deprivation and sudden infant death syndrome. AB - This research uses a case-control study to examine the relationship between deprivation of area of residence and incidence of sudden infant death syndrome (SIDS) in New Zealand. Three hundred sixteen cases and 1221 controls were elicited with geocodable addresses. Infants living in the most deprived areas had a 5.9- (95%CI = 2.9, 12.2) fold increased risk of SIDS compared with those infants living in the least deprived areas. After adjusting for individual-level factors there was no remaining area-level effect. Nearly 27% of control infants lived in the two lowest deciles of deprivation, and almost half of SIDS deaths occurred in these areas. Incidence of SIDS is strongly associated with both deprivation of area of residence and individual socioeconomic factors. This indicates the importance of reaching parents in deprived areas, however, given that approximately 50% of SIDS deaths occurred in areas other than the two most deprived, we recommend that SIDS health messages be delivered to the general population. PMID- 10817478 TI - Increasing mortality differentials by residential area level of poverty: Britain 1981-1997. AB - This paper considers mortality for ages 15-64 for the period 1981-97 in Britain according to population deciles defined by an area-based poverty measure. Over the study period there has been a polarization of life chances such that by 1994 97 almost one quarter of deaths in this age group can be attributed to unfavourable socioeconomic circumstances. Mortality differentials according to socioeconomic circumstances increased in tandem with increases in income inequality. A commitment to redistributive social policies is necessary if the trend of increasing inequality is to be reversed. PMID- 10817479 TI - Surveillance for adverse events associated with anthrax vaccination--U.S. Department of Defense, 1998-2000. AB - Concerns about the potential use of anthrax as a biologic weapon prompted the U.S. Department of Defense (DoD) to announce on December 15, 1997, anthrax vaccination of all U.S. military personnel. This effort is coordinated by the Anthrax Vaccine Immunization Program (AVIP). AVIP plans a phased vaccination process to achieve total force protection against anthrax by 2004. The current phase of implementation includes vaccination of all service members and mission essential DoD civilian employees assigned or deployed to high-threat areas. On the basis of program monitoring, as of April 12, 2000, 425,976 service members had received 1,620,793 doses of anthrax vaccine adsorbed (AVA) (Bioport, Inc., Lansing, Michigan). Some service members have cited concerns about vaccine safety and efficacy in their decision to refuse vaccination, despite the possibility of administrative or disciplinary actions. To assess anthrax vaccination safety, DoD has conducted surveys of vaccinated personnel. This report describes three completed or ongoing surveys. The findings indicate that rates of local reactions were higher in women than men and that no patterns of unexpected local or systemic adverse events have been identified. PMID- 10817480 TI - Serogroup W-135 meningococcal disease among travelers returning from Saudi Arabia -United States, 2000. AB - On April 9, 2000, CDC was notified by national public health agencies in several European countries of cases of serogroup W-135 meningococcal disease among pilgrims returning from the Hajj in Mecca and their close contacts. As of April 20, 2000, the New York City Department of Health had reported three cases of serogroup W-135 meningococcal disease in the United States. PMID- 10817481 TI - Alcohol policy and sexually transmitted disease rates--United States, 1981-1995. AB - In the United States, adolescents and young adults are at higher risk for acquiring sexually transmitted diseases (STDs) than older adults. In addition, young persons who drink alcohol may be more likely than persons who abstain to participate in high-risk sexual activity, such as unprotected sexual intercourse or multiple sexual partners. If alcohol consumption promotes risky sexual behavior (disinhibition caused by the effects of alcohol), state government alcohol policies, such as alcohol taxation and minimum legal drinking age requirements, might reduce STD incidence among adolescents and young adults. Higher alcohol taxes and increases in the minimum legal drinking age have been associated with lower incidences of adverse alcohol-related health outcomes (e.g., motor-vehicle crash-related deaths, liver cirrhosis, suicide, and violent crime, including domestic violence). This report summarizes the findings of a study that suggest higher alcohol taxes and higher minimum legal drinking ages are associated with lower STD incidence among certain age groups. PMID- 10817482 TI - Progress toward global poliomyelitis eradication, 1999. AB - In 1988, the World Health Assembly resolved to eradicate poliomyelitis globally by the end of 2000. Since then, substantial progress has been made in implementing polio eradication strategies, and during 1999 these activities were accelerated to reach the global target. The number of countries where polio is endemic decreased, and the number and quality of vaccination rounds increased. Acute flaccid paralysis (AFP) surveillance improved, and political commitment and the global partnership for polio eradication strengthened. This report updates progress toward achieving the polio eradication goal during 1999. PMID- 10817483 TI - Surveillance for Lyme disease--United States, 1992-1998. AB - PROBLEM/CONDITION: Lyme disease is caused by infection with the spirochete Borrelia burgdorferi and is the most commonly reported vector-borne disease in the United States. Borrelia burgdorferi is transmitted to humans by infected Ixodes scapularis and I. pacificus ticks. Lyme disease is typically evidenced in its early stage by a characteristic rash (erythema migrans), accompanied by nonspecific symptoms (e.g., fever, malaise, fatigue, headache, myalgia, and arthralgia). Lyme disease can usually be treated successfully with standard antibiotic regimens. REPORTING PERIOD: 1992-1998. DESCRIPTION OF SYSTEM: Lyme disease surveillance data are reported to CDC through the National Electronic Telecommunication System for Surveillance, a computerized public health database for nationally notifiable diseases. During 1992-1998, data regarding reported cases of Lyme disease included county and state of residence, age, sex, and date of onset. Descriptive analyses were performed, and cumulative incidence by state, county, age group, and sex were calculated. RESULTS: During 1992-1998, a total of 88,967 cases of Lyme disease was reported to CDC by 49 states and the District of Columbia, with the number of cases increasing from 9,896 in 1992 to 16,802 in 1998. A total of 92% of cases was reported from eight northeastern and mid Atlantic states and two north-central states. Children aged 5-9 years and adults aged 45-54 years had the highest mean annual incidence. INTERPRETATION: Lyme disease is a highly focal disease, with the majority of reported cases occurring in the northeastern and north-central United States. The number of reported cases of Lyme disease increased during 1992-1998. Geographic and seasonal patterns of disease correlate with the distribution and feeding habits of the vector ticks, I. scapularis and I. pacificus. PUBLIC HEALTH ACTION: The results presented in this report will help clinicians evaluate the prior probability of Lyme disease and provide the framework for targeting human Lyme disease vaccine use and other prevention and treatment interventions. PMID- 10817484 TI - Surveillance for influenza--United States, 1994-95, 1995-96, and 1996-97 seasons. AB - PROBLEM/CONDITION: Influenza epidemics occur nearly every year during the winter months and are responsible for substantial morbidity and mortality in the United States, including an average of approximately 114,000 hospitalizations and 20,000 deaths per year. REPORTING PERIOD: This report summarizes U.S. influenza surveillance data from October 1994 through May 1997, from both active and passive surveillance systems. DESCRIPTION OF SYSTEM: During the period covered, CDC received weekly reports from October through May from a) state and territorial epidemiologists on estimates of local influenza activity, b) approximately 140 sentinel physicians on their total number of patient visits and the number of cases of influenza-like illness (ILI), and c) approximately 70 World Health Organization (WHO) collaborating laboratories in the United States on weekly influenza virus isolations. WHO collaborating laboratories also submitted influenza isolates to CDC for antigenic analysis. Throughout the year, vital statistics offices in 121 cities reported deaths related to pneumonia and influenza (P&I) weekly, providing a measure of the impact of influenza on mortality. RESULTS: During the 1994-95 influenza season, 25 state epidemiologists reported regional or widespread activity at the peak of the season. Cases of ILI reported by sentinel physicians exceeded baseline levels for 4 weeks, peaking at 5%. Influenza A(H3N2) was the most frequently isolated influenza virus type/subtype. The longest period of sustained excess mortality was 5 consecutive weeks, when the percentage of deaths attributed to P&I exceeded the epidemic threshold, peaking at 7.6%. During the 1995-96 season, 33 state epidemiologists reported regional or widespread activity at the peak of the season. ILI cases exceeded baseline levels for 5 weeks, peaking at 7%. Influenza A(H1N1) viruses predominated, although influenza A(H3N2) and influenza B viruses also were identified throughout the United States. P&I mortality exceeded the epidemic threshold for 6 consecutive weeks, peaking at 8.2%. The 1996-97 season was the most severe of the three seasons summarized in this report. Thirty-nine state epidemiologists reported regional or widespread activity at the peak of the season. ILI reports exceeded baseline levels for 5 consecutive weeks, peaking at 7%. The proportion of respiratory specimens positive for influenza peaked at 34%, with influenza A(H3N2) viruses predominating. Influenza B viruses were identified throughout the United States, but only one influenza A(H1N1) virus isolate was reported overall. The proportion of deaths attributed to P&I exceeded the epidemic threshold for 10 consecutive weeks, peaking at 9.1%. INTERPRETATION: Influenza A(H1N1), A(H3N2), and B viruses circulated during 1994-1997. Local surveillance data are important because of geographic and temporal differences in the circulation of influenza types/subtypes. PUBLIC HEALTH ACTIONS: CDC conducts active national surveillance annually from October through May for influenza to detect the emergence and spread of influenza virus variants and monitor the impact of influenza-related morbidity and mortality. Surveillance data are provided weekly throughout the influenza season to public health officials, WHO, and health-care providers and can be used to guide prevention and control activities, vaccine strain selection, and patient care. PMID- 10817485 TI - Attention-deficit/hyperactivity disorder: a neuropsychiatric disorder with childhood onset. AB - Attention-deficit/hyperactivity disorder (ADHD) is characterized by attentional problems such as hyperkinesia, restlessness and disturbances in timing. Environmental psychosocial factors interact with a genetic predisposition in causing measurable biological impairment. It is a common, persistent disorder of childhood that may change in manifestation with development from preschool through adult life. Untreated, it predisposes children to psychiatric and social pathology later in life. Unlike other psychiatric disorders of childhood, it can be successfully treated. PMID- 10817486 TI - Multiple symmetric lipomatosis: an unusual cause of childhood obesity and mental retardation. AB - Multiple symmetric lipomatosis (MSL), also known as Launois-Bensaude syndrome or Madelung's disease, is a rare disorder predominantly seen in middle-aged male patients. The disorder is characterized by large subcutaneous fat masses distributed around the neck, shoulders, and other parts of the trunk, often associated with nervous system abnormalities. A close relationship to alcoholism, metabolic disturbances and malignant tumours has been observed. Until now, MSL has only been described in adults. We report on the first two children, a 9-year old girl and a 13-year-old boy, with the characteristic clinical findings of MSL. The girl presented with severe obesity, developmental delay, mild mental retardation, peripheral neuropathy, and latent hypothyroidism. In addition, she had elevated lactate concentrations in blood and cerebral spinal fluid suggesting mitochondrial dysfunction. Biochemical analyses of muscle showed a respiratory chain complex II deficiency. The boy suffered from severe obesity, mild mental retardation and insulin resistant diabetes mellitus. In both children, analyses of the mitochondrial genome did not reveal major deletions nor the MERRF 8344 point mutation. MSL seems to be a new neurometabolic disorder with heterogeneous clinical expression whose pathogenesis is still unknown. PMID- 10817487 TI - Diaphragmatic spinal muscular atrophy with bulbar weakness. AB - We present the clinical and histopathological features of a child affected by diaphragmatic spinal muscular atrophy. The child was born with mild distal arthrogryposis, mild hypotonia and developed marked diaphragmatic and bulbar muscle weakness in the first week of life. Electrophysiological and pathological investigations performed at presentation were not conclusive, while the investigations performed at 3 months showed a clear neurogenic picture. Genetic studies excluded involvement of the SMN gene, or of other genes located on chromosome 5q, confirming that this syndrome represents a different entity from typical proximal spinal muscular atrophy. PMID- 10817488 TI - Velocardiofacial syndrome associated with atrophy of the shoulder girdle muscles and cervicomedullary narrowing. AB - Velocardiofacial syndrome is the most common microdeletion syndrome in humans. It is secondary to a chromosome 22q11 rearrangement and is characterized by craniofacial abnormalities, heart defects and learning disability. We report a case of a 10-year-old girl with a chromosome 22q11 deletion who, in addition to learning difficulties, hypernasal speech and mild dysmorphic features, had weakness and wasting of the shoulder girdle muscles but no cardiac involvement. Brain magnetic resonance imaging revealed narrowing of the cervicomedullary junction. The clinical features of this patient with velocardiofacial syndrome further expand the spectrum of abnormalities associated with this condition. PMID- 10817489 TI - An unusual case of hyperekplexia. AB - Hyperekplexia is a rare paroxysmal disorder characterized by exaggerated startle response, hypertonia during infancy and a transient increase in tone following startle attacks. We report an unusual case of hyperekplexia in a young man. In addition to common symptoms of the condition, we found generalized spasticity persisting beyond infancy, and facial and skeletal dysmorphism. Because of an unsteady gait with frequent falls and raised serum creatine kinase levels, a congenital myopathy had been suspected in the past and an abnormal muscle biopsy had been documented. We diagnosed hyperekplexia at the age of 21 years on clinical grounds and following the response to pharmacological treatment. A mutation in the alpha1 subunit of the glycine receptor confirmed the diagnosis. A repeated needle muscle biopsy demonstrated mild myopathic changes, which we considered to be secondary to increased muscle tone. This case highlights the diagnostic difficulties of hyperekplexia, particularly in sporadic cases with unusual presentation. PMID- 10817490 TI - Landau-Kleffner syndrome with onset at 18 months and an initial diagnosis of pervasive developmental disorder. AB - We report one of the youngest and most intensively studied cases of Landau Kleffner syndrome, with a follow-up of 5 years. The boy developed normally until the age of 18 months when he had two attacks, possibly epileptic. Electroencephalogram (EEG) was normal. Over the next 5 months he lost his six to ten words, did not engage with other children and became mute. When he was 34 months old a child-psychiatrist suggested a diagnosis of pervasive developmental disorder or developmental dysphasia. An EEG 3 months later showed abnormalities typical of Landau-Kleffner syndrome. His non-verbal abilities were normal as well as his neurological examination and brain magnetic resonance imaging (MRI). A trial of clobazam and vigabatrin was unsuccessful. When he was 4 years and 9 months old he was treated with corticosteroids and within 3 months his vocabulary increased from the standard for 1 1/2 years of age to that for 2 1/2 years of age. His language abilities continued to improve slowly until a stagnation period at the age of 6 years and 9 months. A second course of corticosteroids improved his comprehension and vocabulary to an almost normal level, and his EEG normalized. A total of 11 EEGs were obtained; all included sleep, but continuous spike and wave during slow sleep was never documented. This report illustrates that Landau-Kleffner syndrome should be considered as an alternative diagnosis in children diagnosed with developmental dysphasia. An EEG including sleep should be considered, and in the presence of abnormalities a trial of anti-epileptic drugs or corticosteroids should be undertaken. PMID- 10817491 TI - Gene table: epilepsy (update). PMID- 10817492 TI - Rett syndrome: genetic horizons. Report on a satellite symposium at the 3rd Congress of the European Paediatric Neurology Society Nice, November, 1999. PMID- 10817493 TI - Cancer vaccines. PMID- 10817494 TI - Can biological markers improve the management of breast cancer patients? Biomarkers Ad-hoc Group of the United Kingdom Coordinating Committee on Cancer Research. PMID- 10817495 TI - Biological markers: maintaining standards. Biomarkers Ad-hoc Group of the United Kingdom Coordinating Committee on Cancer Research. PMID- 10817496 TI - Concentrations of tamoxifen and its major metabolites in hormone responsive and resistant breast tumours. AB - Patients treated with tamoxifen (TAM) for primary breast cancer often manifest de novo or acquired resistance, possibly through changes in drug metabolism. Using solid-phase extraction methods and reversed-phase high-performance liquid chromatography separations, levels of TAM and metabolites 4-hydroxytamoxifen (4OH) and desmethyltamoxifen (DMT) have been measured in plasma and tumour tissue from breast cancer patients treated with TAM for at least 3 months. Patients were categorized into those with tumours responding to TAM and those showing de novo or acquired resistance. Levels of TAM, 4OH and DMT in both plasma and tissue samples were correlated with clinical response, length of treatment and patient weight. Interesting results included accumulation of 4OH in tumour tissues over time in all patients, with significance reached in the acquired resistance group. In addition, significantly lower levels of 4OH and DMT were found in plasma taken from responding patients after 3 months of treatment when compared to non responding patients, and a small group of ER-poor patients showed significantly lower levels of all three species in plasma when compared to other patients. Whilst not explaining TAM resistance in all cases, these differences could account for the development of resistance to TAM treatment in certain subgroups of patients. PMID- 10817497 TI - Risk factors for nephrotoxicity after ifosfamide treatment in children: a UKCCSG Late Effects Group study. United Kingdom Children's Cancer Study Group. AB - The aim of this multicentre study was to document the nephrotoxicity associated with ifosfamide and evaluate risk factors in 148 children and young people with sarcomas who underwent investigation of renal function on one occasion each, at a median of 6 (range 1-47) months after completion of ifosfamide (median dose 62.0 (range 6.1-165.0) g/m2). Investigations included glomerular filtration rate (GFR), serum bicarbonate (HCO3) and phosphate (PO4), and renal tubular threshold for phosphate (Tmp/GFR). A clinically relevant nephrotoxicity score' was derived. GFR was < 90 ml/min/1.73 m2 in 61 of 123 evaluable patients, Tmp/GFR < 0.9-1.1 mmol/l (age-dependent) in 45/103, serum PO4 < 0.9-1.mmol/l (age-dependent) in 28/135, and serum HCO3 < 20 (< 18 in infants) mmol/l in 22/95. Of 76 fully evaluable patients: 50% had mild, 20% moderate and 8% severe nephrotoxicity. Higher total ifosfamide dose correlated significantly with greater glomerular and tubular toxicity (P < 0.01); other risk factors, including age at treatment, demonstrated no consistent significant independent effect. Chronic ifosfamide related glomerular and proximal tubular toxicity were common in this large comprehensive study. Restriction of total ifosfamide dose to < 84 g/m2 will reduce the frequency of, but not abolish, clinically significant nephrotoxicity, whilst doses > 119 g/m2 are associated with a very high risk of severe toxicity. PMID- 10817498 TI - Polymorphisms in GSTM1, GSTT1 and CYP1A1 and risk of pancreatic adenocarcinoma. AB - A prospective study of 149 unselected incident cases of pancreatic adenocarcinoma and 146 ethnically-matched controls found no associations between GSTM1 (adjusted odds ratio (AOR) 1.14), GSTT1 (AOR: 1.19) and CYP1A1 (AOR: 1.08) polymorphisms and pancreatic cancer susceptibility. Smoking and drinking status did not affect results. These polymorphisms do not appear to be important gene modifiers in pancreatic cancer. PMID- 10817499 TI - Molecular detection of thyroglobulin mRNA transcripts in peripheral blood of patients with thyroid disease by RT-PCR. AB - The sensitive detection of circulating tumour cells in patients with differentiated thyroid cancer may precede the detection of relapse by other diagnostic studies--such as serum thyroglobulin-and thus may have important therapeutic and prognostic implications. We performed reverse transcription polymerase chain reaction (RT-PCR) on blood samples from patients diagnosed with thyroid disease using two different RT-PCR sensitivities. Additionally, tissue specificity of TG mRNA-expression was determined using RNA extracts from 27 different human tissues. The lower limit of detection was 50-100 TG mRNA producing cells/ml blood using a 'normal' RT-PCR sensitivity and 10-20 cells/ml blood using a 'high' sensitivity. With the normal sensitivity TG mRNA was detected in 9/13 patients with thyroid cancer and metastasis, 63/137 patients with a history of thyroid cancer and no metastasis, 21/85 with non-malignant thyroid disease and 9/50 controls. With the high sensitivity TG mRNA was detected in 11/13 patients with thyroid cancer and metastasis, 111/137 patients with a history of thyroid cancer and no metastasis, 61/85 with non-malignant thyroid disease and 41/50 controls. Interestingly, using the normal RT-PCR sensitivity TG mRNA transcripts are specific for thyroid tissue and detectable in the peripheral blood of controls and patients with thyroid disease, which correlates with a diagnosis of metastasized thyroid cancer. However, with a high RT-PCR sensitivity, TG mRNA expression was found not to be specific for thyroid tissue and was not correlated with a diagnosis of thyroid cancer in patients. As a consequence, to date TG mRNA detected by RT-PCR in the peripheral blood cannot be recommended as a tumour marker superior to TG serum-level. PMID- 10817500 TI - Morphometric grading of breast cancer: thresholds for tubular differentiation. AB - We evaluated the degree of tubular differentiation in 172 samples of invasive ductal breast cancer in order to determine numerical thresholds for histological breast cancer grading. The tubular differentiation in each sample was defined as the fraction of fields showing tubular differentiation (FTD). The analysis was based on Kaplan-Meier curves reflecting survival and recurrence of disease, univariate and multivariate analyses of Cox's regression, and maximum efficiencies of ROC analysis. The minimum P-value cut-off for FTD was determined at 59%. The practical interpretation is that tubular differentiation in the neoplasm observed in at least 60% of microscopical fields in the tumour area indicates favourable prognosis of disease. The relative risks for breast cancer death for patients with FTD below 59% as compared with those with FTD above 59% were 6.7--and 6.3-fold (univariate and multivariate analyses respectively). Another threshold could be determined at FTD 23%, although this threshold was associated with clearly lower statistical significancies. The paper introduces two possible solutions for application of the thresholds to the morphometric breast cancer grading system. The study also emphasizes the clinical relevance of the evaluation of tubular differentiation in breast cancer. The consistent morphometric evaluation method was vital in allowing the full weight of the biological significance of tubular differentiation to emerge. PMID- 10817501 TI - nm23-H1 expression defines a high-risk subpopulation of patients with early-stage epithelial ovarian carcinoma. AB - The role of the nm23 gene in human ovarian cancer is still controversial. We studied the expression of the nm23-H1 gene in 247 human epithelial ovarian carcinomas. The patients were followed-up until their death, or for a minimum of 5 years if they survived. The expression of the gene was studied by means of immunohistochemistry and a semiquantitative scoring system considering the staining intensity and the number of reactive tumour cells. Patients carrying tumours with higher expression scores (4-6 on a scale from 0 to 6) had a significantly lower survival (P = 0.01) than the rest. Further stratified statistical analysis revealed that this effect was mainly attributable to the subgroup of patients with early-stage (I and II), well- and moderately differentiated tumours. In fact, a multivariate analysis carried out for this subset of patients showed nm23-overexpression to be the only significant independent predictor of an ominous prognosis. The association of nm23 overexpression with a worse prognosis was most probably not due to mutation of the nm23 gene, since mutational analysis in 60 tumours by means of single-strand conformational polymorphism and direct sequencing disclosed only one mutation, which was located outside the open reading frame. Our results seem to indicate that nm23 expression is associated with a significantly worse prognosis in early stage, well-differentiated epithelial ovarian carcinoma, a finding with important clinical implications, considering that many patients with ovarian cancers showing these features do not undergo any further treatment beyond surgical staging. If confirmed, they could help in tailoring the treatment of these patients in the future. PMID- 10817502 TI - The expression profile for the tumour suppressor gene PTEN and associated polymorphic markers. AB - PTEN, a putative tumour suppressor gene associated with prostate and other cancers, is known to be located within the chromosomal region 10q23.3. Transcription of the PTEN gives rise to multiple mRNA species. Analyses by Northern blots, using cell lines which express PTEN together with cell lines which have lost the PTEN or carry a truncated version of the gene, has allowed us to demonstrate that the pseudogene is not transcribed. In addition, 3' RACE studies confirmed that the multiple mRNA species arising from the gene probably result from the use of alternative polyadenylation sites. No evidence for tissue- or cell-specific patterns of transcription was found. Analysis by 5' RACE placed the putative site for the start of transcription around 830 bp upstream of the start codon. A map of the location of the PTEN gene with a series of overlapping YAC, BAC and PACs has been constructed and the relative position of eight microsatellite markers sited. Two known and one novel marker have been positioned within the gene, the others are in flanking regions. The more accurate location of these markers should help in future studies of the extent of gene loss. Several polymorphisms were also identified, all were within introns. Four of the common polymorphisms appear to be linked. In blood, DNA from 200 individuals, including normal, BPH and prostate cancer patients, confirmed this link. Only two samples of 200 did not carry the linked haplotype, both were patients with advanced prostate cancer. It is possible that such rearrangements within PTEN could be evidence of predisposition to prostate cancer in this small number of cases. PMID- 10817503 TI - High incidence of SV40-like sequences detection in tumour and peripheral blood cells of Japanese osteosarcoma patients. AB - Recent studies have revealed the evidence for the significance of SV40 genome in human malignancies. In this paper, the presence of SV40-like sequences was investigated in 54 Japanese osteosarcomas in which mutations of the retinoblastoma (Rb), p53, MDM2, and CDK4 genes had been already analysed. Using polymerase chain reaction and Southern hybridization, SV40-like sequences were detected in 25 cases (46.3%). In most cases, only a part of SV40 genome was detected, and the regulatory region containing enhancer sequences was most frequently found (21/54, 38.9%). There was no apparent relationship between the presence of SV40-like sequences and tumour suppressor genes mutations in each tumour. The SV40-like sequences were also detected in peripheral blood cells of substantial proportion of the patients (43.3%), whereas the incidence was much lower (4.7%) in normal healthy controls. This difference is statistically highly significant (P < 0.0001), suggesting that the presence of SV40-like sequences, even if only a part, may play some roles to predispose individuals to osteosarcoma. PMID- 10817505 TI - Nuclear translocation of beta-catenin in colorectal cancer. AB - Post-translational stabilization of beta-catenin through mutation of the adenomatous polyposis coli (APC) gene has been proposed as an early step in colorectal carcinogenesis. Beta-catenin may translocate from the cytoplasm to the nucleus, where it might serve as a transcriptional factor to stimulate tumour formation. We investigated intracellular localization of beta-catenin in sporadic colorectal adenomas and cancers as well as familial adenomatous polyposis (FAP). Nuclear over-expression of beta-catenin was observed in 35% (7/20) of intramucosal cancers and 42% (23/55) of invasive cancers but was not seen in any adenomas from sporadic or FAP cases. Cytoplasmic beta-catenin in adenomas was significantly higher than that of normal mucosa in both sporadic and FAP cases. The cytoplasmic intensity index of cancers was significantly higher than that of sporadic adenomas, but the index was not correlated with nuclear expression in cancers. These findings suggest that nuclear translocation of beta-catenin is involved in development of intramucosal cancer rather than adenoma, independent of APC mutations. Cytoplasmic accumulation of beta-catenin may occur in adenomas, but it remains to be determined whether this is a cause or a consequence of colorectal cancer. PMID- 10817504 TI - Quantitation of Fas and Fas ligand gene expression in human ovarian, cervical and endometrial carcinomas using real-time quantitative RT-PCR. AB - Alterations in the expression of Fas (CD95/APO-1) and its ligand (FasL) have been demonstrated in various types of cancers as a mechanism for tumour cell to escape from the immune system. In the present study, we evaluated the expression of the Fas and FasL genes in a wide range of primary gynaecological carcinomas. These included 31 ovarian, 29 cervical and 25 endometrial carcinoma tissues as well as four ovarian and three cervical carcinoma cell lines. Our real-time quantitative reverse transcription polymerase chain reaction analysis revealed that down regulation of Fas expression is more prominent than the up-regulation of FasL expression in all types of gynaecological cancer studied. This down-regulation of Fas expression was also true for the seven carcinoma cell lines. Only one cervical carcinoma cell line, DoT, exhibited a high level of FasL expression. These results indicated that down-regulation of Fas expression is a common abnormality in many types of cancers including gynaecological cancers, whereas an increase in FasL expression is not a common phenomenon in these cancers. PMID- 10817506 TI - Angiogenically active vascular endothelial growth factor is over-expressed in malignant human and rat prostate carcinoma cells. AB - Vascular endothelial growth factor (VEGF) is one of the most potent factors for stimulating angiogenesis, an essential process required for expansion of primary tumour and dissemination of malignant cells. To investigate the possible role of VEGF in facilitating metastasis of prostate cancer via stimulating angiogenesis, we have used Northern and slot blotting, reverse transcription polymerase chain reaction, nucleotide sequence analysis and enzyme-linked immunosorbent assay to compare the VEGF expression in series of human and rat cell lines with either benign or malignant characteristics. We have also employed the chick chorioallantoic membrane (CAM) assay to measure the angiogenic activity of the VEGF derived from both benign and malignant cells. The level of VEGF mRNA expressed in the seven malignant human and rat cell lines is 3.5- to 10-fold higher than that expressed in the benign cell lines. The three metastatic variants, generated by transfection of a benign cell line with DNA extracted from prostate carcinoma cells, expressed 2.5 to 5 times more VEGF mRNA than their parental benign cells. While VEGF 121 and 165 were predominantly expressed by both the benign and malignant cells, the transcript representing VEGF 189 isoform was only detected in the malignant cells. At protein level, three human malignant cell lines produced more VEGF (2.7-7.9 ng ml(-1)) than the benign cell line (1.3 ng ml(-1)). CAM assay detected a VEGF-dependent angiogenic activity in the medium from malignant cells, but only a relatively weak VEGF-independent activity in the medium from benign cells. These results demonstrated that malignant cells did over-express VEGF and only the VEGF derived from malignant cells was angiogenically active. Thus, we suggest that the VEGF produced by malignant cells might play an important role in facilitating metastasis of prostatic cancer. PMID- 10817507 TI - Plasminogen activator inhibitor 1 may promote tumour growth through inhibition of apoptosis. AB - Plasminogen activator inhibitor 1 (PAI-1) has been found to be a bad prognostic factor in a number of tumours but the reason has not been fully explained. The human prostate cancer cell line PC-3 and the human promyelocytic leukaemia cell line HL-60 were used in this study to determine the effect of PAI-1 on spontaneous and induced apoptosis in culture. Apoptosis was induced with camptothecin or etoposide. Addition of a stable variant of PAI-1 or wild-type PAI 1 to these cells resulted in a significant inhibition of apoptosis. In contrast, both the latent form of PAI-1 and the stable variant of PAI-1 inactivated by a specific neutralizing monoclonal antibody, or the stable variant of PAI-1 in a complex with recombinant urokinase did not inhibit apoptosis. This indicated that the inhibitory activity of PAI-1 was required for its anti-apoptotic effect but the urokinase-type plasminogen activator receptor was not involved. These findings provide an explanation for the bad prognostic correlation of high PAI-1 levels in tumours. The anti-apoptotic effect was also found in non-tumoural cells including human umbilical vein endothelial cells and the benign human breast epithelial cell line MCF-10A, suggesting that this is a novel physiologic function of PAI-1. PMID- 10817508 TI - Inhibition of radiation-induced apoptosis by dexamethasone in cervical carcinoma cell lines depends upon increased HPV E6/E7. AB - Through a glucocorticoid-responsive promoter, glucocorticoids can regulate the transcription of the human papillomavirus (HPV) E6 and E7 viral genes which target the tumour suppressor proteins p53 and Rb respectively. In C4-1 cells, the glucocorticoid dexamethasone up-regulated HPV E6/E7 mRNA and decreased radiation induced apoptosis. In contrast, dexamethasone had no effect on apoptosis of cells that either lack the HPV genome (C33-a) or in which HPV E6/E7 transcription is repressed by dexamethasone (SW756). Irradiated C4-1 cells showed increased p53 expression, while dexamethasone treatment prior to irradiation decreased p53 protein expression. In addition, p21 mRNA was regulated by irradiation and dexamethasone in accordance with the observed changes in p53. Overall, glucocorticoids decreased radiation-induced apoptosis in cervical carcinoma cells which exhibit increased HPV E6/E7 transcription and decreased p53 expression. Therefore, in HPV-infected cervical epithelial cells, p53-dependent apoptosis appears to depend upon the levels of HPV E6/E7 mRNA. PMID- 10817509 TI - Regulatory effect of interleukin-4 and interleukin-13 on colon cancer cell adhesion. AB - To assess the role of interleukin-4 (IL-4) and interleukin-13 (IL-13) in colon cancer cell-cell adhesion, we investigated the effect of both cytokines in human colon cancer cell line, colo205 cell-cell adhesion. IL-4 receptor was expressed on the cell surface of colo205, and recombinant IL-4 inhibited colo205 cell-cell adhesion in a dose-dependent fashion without inhibiting cell proliferation. Flow cytometric analysis revealed that monoclonal antibodies (mAbs) directed against E cadherin and carcinoembryonic antigen (CEA) inhibited colo205 cell-cell adhesion and IL-4 significantly inhibited the expression of E-cadherin and CEA. IL-13 also inhibited colo205 cell-cell adhesion. These results indicated that IL-4 and IL-13 inhibited colon cancer cell-cell adhesion by down-regulation of E-cadherin and CEA molecules. We then investigated the expression of both cytokines from freshly isolated colon cancer tumour-infiltrating lymphocytes (TILs). With reverse transcription-polymerase chain reaction and flow cytometric analysis, we demonstrated that colon TILs expressed IL-4 and IL-13 mRNA and protein. These results suggest that Th 2 type cytokines IL-4 and IL-13 locally-produced from TILs may regulate colon cancer adhesion by down-regulation of adhesion molecules. PMID- 10817511 TI - Isolation and characterization of an IGROV-1 human ovarian cancer cell line made resistant to Ecteinascidin-743 (ET-743). AB - By exposing Igrov-1 human ovarian cancer cells to increasing concentrations of Ecteinascidin-743 (ET-743), either for a short or prolonged time, we obtained sublines resistant to ET-743 which overexpress Pgp. The most resistant clone (Igrov-1/25 ET) was evaluated for biological and pharmacological characterizations. The increased Pgp levels of Igrov-1/25 ET were not due to amplification of the mdr-1 gene but to increased mRNA levels. No increase in other multidrug resistance-related proteins such as MRP or LRP was observed in Igrov-1/25 ET. The IC50 values of ET-743 against Igrov-1/25 ET was approximately 50 times higher than the parental cell line. Resistance was not reversed while maintaining the cell line in drug-free medium for at least 24 months. Igrov-1/25 ET was cross-resistant to Doxorubicin and VP16 while it was equally sensitive to L-PAM, MNNG, CPT and only marginally less sensitive to Cis-DDP and Oxaliplatin compared to the parental cell line. Igrov-1/25 ET exposed to Doxorubicin retained this drug much less, mainly because of a more efficient drug efflux. The cyclosporine analogue SDZ PSC-833 reversed the resistance of Igrov-1/25 ET to ET 743, without any enhancement of the drug activity against the parental Igrov-1 cell line. Igrov-1/25 ET exhibits typical features of cell lines overexpressing the mdr-1 gene and can be a potentially useful tool in selecting ET-743 non-cross resistant analogues as well as to investigate methods to counteract resistance to this drug. PMID- 10817510 TI - Antagonists of growth hormone-releasing hormone (GH-RH) inhibit IGF-II production and growth of HT-29 human colon cancers. AB - Insulin-like growth factors (IGFs) I and II are implicated in progression of various tumours including colorectal carcinomas. To interfere with the production of IGFs, we treated male nude mice bearing xenografts of HT-29 human colon cancer with various potent growth hormone-releasing hormone (GH-RH) antagonists. Twice daily injections of antagonist MZ-4-71, 10 microg intraperitoneally or 5 microg subcutaneously (s.c.) resulted in a significant 43-45% inhibition of tumour growth. Longer acting GH-RH antagonists, MZ-5-156 and JV-1-36 given once daily at doses of 20 microg s.c. produced a 43-58% decrease in volume and weight of cancers. Histological analyses of HT-29 cancers demonstrated that both a decreased cell proliferation and an increased apoptosis contributed to tumour inhibition. GH-RH antagonists did not change serum IGF-I or IGF-II levels, but significantly decreased IGF-II concentration and reduced mRNA expression for IGF II in tumours. In vitro studies showed that HT-29 cells produced and secreted IGF II into the medium, and addition of MZ-5-156 dose-dependently decreased IGF-II production by about 40% as well as proliferation of HT-29 cells. Our studies demonstrate that GH-RH antagonists inhibit growth of HT-29 human colon cancers in vivo and in vitro. The effect of GH-RH antagonists may be mediated through a reduced production and secretion of IGF-II by cancer cells. PMID- 10817512 TI - Involvement of energy metabolism in the production of 'bystander effects' by radiation. AB - These experiments were done to determine if interference with energy metabolism and REDOX biochemistry during low LET radiation exposure would alter the ability of medium harvested from the irradiated cells to induce a bystander effect in unirradiated cells. Human keratinocyte cells and CHO-K1 mutant cell lines were irradiated using cobalt 60. Clonogenic assays were used to determine the reproductive death of the cells exposed to direct irradiation or medium from irradiated cells. The persistence in progeny was also examined. Use of apoptosis inhibitors or medium from the LDH or G6PD null cell lines, reduced or prevented the bystander effect. Transfection with G6PD recovered the effect. Treatment with anti-oxidant substances, L-lactate and L-deprenyl prevented bystander factor associated cell kill. The lactate analogue, oxamate, was less effective. Data from experiments where media harvested from the different cell lines was exchanged suggest that signal production and cellular response may involve different mechanisms. The effects on exposed cells were transmitted to progeny which also showed excessive levels of cell death for several generations. The results suggest that energy/REDOX metabolism may be involved in the expression of a radiation induced bystander response. Given the aberrant energy metabolism in tumour cells, this may have implications for dose escalation in radiotherapy. PMID- 10817513 TI - Relevance of proliferative and pro-apoptotic factors in non-small-cell lung cancer for patient survival. AB - This investigation first set out to analyse which cellular proliferative and apoptotic factors, in addition to the clinical prognostic factors, are most predictive in patients with non-small-cell lung carcinomas (NSCLC). To this purpose, we related the proliferative factors proliferating cell nuclear antigen (PCNA), cyclin A, cyclin D1, cyclin-dependent kinase 2 (cdk2), cdk4 and the proportion of cell cycle phases in NSCLC to the survival times of 150 patients. Additionally, we associated the expressions of Fas, Fas ligand and caspase-3 in NSCLC to patient survival. Immunohistochemistry was used to determine the proteins and flow cytometry to assess the proportion of cell cycle phases. Patients with PCNA-positive carcinomas had significantly shorter survival times than patients with PCNA-negative carcinomas (median survival times: 51 vs 89 weeks). Corresponding results were obtained with the factor cyclin A (64 vs 92 weeks), with the factor cdk2 (76 vs 89 weeks), with the factor cdk4 (62 vs 102 weeks) and with the proportion of S phases (86 vs 121 weeks). Patients with an expression of the apoptotic factors had a more favourable prognosis than patients with negative carcinomas. The median survival times of cancer patients with Fas expression was 86 weeks and of those without Fas expression only 69 weeks. Corresponding results were obtained with the Fas ligand (87 vs 41 weeks) and caspase 3 (87 vs 34 weeks). In order to determine whether a combination of factors can yield improved prognostic information, we investigated all possible combinations of the proliferative and apoptotic factors. Patients with tumours having a high proliferative activity, but which did not express apoptotic factors had the shortest survival times while patients with a low proliferative activity and a high expression of apoptotic factors had the most favourable outcome. A multivariate analysis (Cox model) of the cellular and clinical prognostic factors indicated that stage, lymph node involvement, Fas, PCNA and cyclin A are the most important prognostic factors for the clinical outcome of patients with non-small cell lung carcinomas. PMID- 10817514 TI - Adjuvant interferon in the treatment of melanoma. PMID- 10817515 TI - Serum tissue polypeptide-specific antigen (TPS): what is its diagnostic value? PMID- 10817516 TI - Genomic arrangement of a putative operon involved in maltose transport in the Mycobacterium tuberculosis complex and Mycobacterium leprae. AB - A Mycobacterium bovis gene coding for a putative MalE maltose binding protein was cloned and its full-length sequence determined. Database searches revealed 99.9% identity with IpqY, encoding a putative sugar uptake protein from Mycobacterium tuberculosis strain H37Rv. The deduced protein product showed high sequence similarity to MalE-like proteins from a variety of bacterial species, including Mycobacterium leprae. Analysis of flanking database sequences from M. tuberculosis and M. leprae revealed the presence of malF-, malG- and malK-like genes. Comparison of these mycobacterial sequences with other maltose operons has allowed us to deduce a unique genomic arrangement of the genes involved in the uptake of maltose in members of the Mycobacterium tuberculosis complex and M. leprae. PMID- 10817517 TI - Genomic variability of hepatitis G virus/GBV-C at the NS3 region: clinical implications. AB - A new hepatitis virus, named GBV-C or hepatitis G virus (HGV), closely related to the hepatitis C virus (HCV), was identified in 1994. The existence of quasispecies in HCV is very important. In this work polymerase chain reaction amplification of the NS3 region of the genome of GBV-C/HGV and heteroduplex mobility assay (HMA) were combined to investigate the presence of quasispecies in patients with chronic infection by GBV-C/HGV. Patients with chronic infection by HCV were used to validate the method. The HMA was also used to investigate the similarity between the cited genomic region of GBV-C/HGV in different infected patients. A high degree of heterogeneity was found for HGV existing as quasispecies and as differences between samples. This is of extreme importance because of the intrinsic clinical and pathogenic implications of quasispecies of a virus capable of producing disease, and is in accord with other studies which report on the genomic variability of the NS3 region. PMID- 10817518 TI - Fatty acids enhanced tubermycin production by Pseudomonas strain 2HS. AB - A new microbial isolate, Pseudomonas 2HS, produced trace amounts of a greenish yellow pigment when grown aerobically in a 1% yeast extract medium at 30 degrees C and shaken at 250 rpm for 5 days. In contrast, cells produced more greenish yellow pigment (2.16 mg/15 ml culture) when grown in the presence of 0.5% 12 hydroxyoctadecanoic acid (w/v). The greenish-yellow pigment was identified as phenazine-1-carboxylic acid (tubermycin B), and the Pseudomonas 2HS was identified as P. aeruginosa 2HS. This is the first report that 12 hydroxyoctadecanoic, ricinoleic and other fatty acids can enhance the production of phenazine-1-carboxylic acid by a Pseudomonas species. PMID- 10817520 TI - Growth of the fungal pathogen Candida in parotid saliva of patients with burning mouth syndrome. AB - Subclinical Candida infection has been suggested as one of the aetiological factors in patients with burning mouth syndrome (BMS). In order to investigate the possible factors which contribute to the relatively high isolation rate of Candida in BMS, parotid saliva samples (20 in toto) from patients with this condition were collected and the growth of Candida in each sample dynamically observed using a computerized turbidometric assay system. A total of thirteen parotid saliva samples obtained from healthy individuals served as normal controls. The results showed no significant growth differential within the test and control saliva samples, when a single isolate each of Candida albicans and Candida tropicalis were cultured for 24 h, at 37 degrees C. A single isolate of Candida glabrata tended to grow better in the saliva from BMS patients than the controls. These results indicate that the composition of saliva may be a contributory factor for the high isolation rate of Candida in saliva of BMS patients. PMID- 10817519 TI - Endotoxin-triggered haematological interactions in Fusobacterium necrophorum infections. AB - The haematological mechanisms in the course of liver abscess formation were evaluated. They were examined by employing viable cells of Fusobacterium necrophorum subsp. necrophorum and Fusobacterium necrophorum subsp. funduliforme in comparison with their endotoxins. Whole cell infection with F.n. necrophorum led to neutrophilia and to a concomitant monocytosis in parallel with those responses induced by the in vivo injection of its endotoxin. Viable infection with F.n. funduliforme was characterized by a sustained endotoxin-related monocytosis against neutropenia. The stimulatory impact of endotoxin on monocytes when released from a viable F.n. funduliforme infection suggested an inherently peculiar mechanism which differed from the induction of both neutrophilia and monocytosis when F.n. funduliforme endotoxin was administered alone. The neutrophilic inducing capacity of the F.n. necrophorum endotoxin was equally illustrated by its positive chemotactic effect on polymorphonuclear neutrophils in vitro. The data presented here emphasize the virulence of F.n. necrophorum viewed in reference to changes in leucocyte trafficking and as complemented by a relatively high endotoxin content. PMID- 10817521 TI - Studies on mevinolin production by some fungi. AB - The potentiality of 25 fungal species, belonging to fourteen genera isolated from Egyptian soils, to produce mevinolin, a hypocholesterolaemic agent, when grown on selected substrates was tested. Samples of culture filtrates were tested by thin layer chromatography and the positive results were further assessed by high pressure liquid chromatography analysis. It was found that nearly one-third of the fungi showed positive results for production of mevinolin. Aspergillus terreus was distinguished by its capacity to produce mevinolin when cultivated on selected media. Some factors influencing mevinolin production and the growth of A. terreus were also studied. The maximal mevinolin yields were achieved after 8 days incubation at 30 degrees C. An initial pH value of 5-6 was the optimum for growth of A. terreus and mevinolin production. PMID- 10817522 TI - Angiogenesis inhibitors. New anticancer strategy. AB - Neoangiogenesis has been proved to be crucial in neoplasmatic tumor growth and metastases. Over the last few years, the factors that have both a positive (angiogenic) and negative (antiangiogenic) influence on tumor growth have been identified. The potential use of natural and synthetic factors that suppress vasculature formation as anticancer drugs is currently under intense investigation. Recently, several antiangiogenic compounds, including TNP-470 or matrix metalloproteinase inhibitors, have entered clinical trials. This review will describe the main groups of angiogenesis inhibitors, their mechanisms of action and some data from clinical studies. PMID- 10817523 TI - Behavioral and memory improving effects of mirtazapine in rats. AB - These experiments examined the effects of the antidepressant mirtazapine in several behavioral and memory tests. The tests were carried out on male Wistar rats weighing about 200 g. The drugs were injected 30 min before the tests. The aim of the locomotor activity test was to select a dose which had no influence on the motility of the animals and, at the same time, was active at least in one behavioral test. The chosen dose was 2.5 mg/kg. In the two-compartment exploratory test, 2.5 mg/kg of mirtazapine had a distinct anxiolytic effect after the first treatment, after 7 days the effect was weaker but still significant and it disappeared after 14 days. In the forced swimming test, the immobility time was shortened only after 14 days of administering the drug. In the maze test, mirtazapine shortened the food finding time (it improved memory) and counteracted memory loss induced by scopolamine. In the conditioned avoidance responses test (CARs), mirtazapine improved memory only after its earlier impairment by scopolamine. The authors cohclude, contrary to some published data, that after proper dose (adequate for other tests but not for the locomotor activity test), mirtazapine has a distinct memory improving activity or a memory restoring effect after scopolamine treatment. PMID- 10817524 TI - Synergistic effect of pramipexole and sertraline in the forced swimming test. AB - The authors studied the effect of sertraline, one of selective serotonin reuptake inhibitors (SSRIs), and pramipexole, administered jointly, to male Wistar rats in the forced swimming test. Both those drugs were injected three times (24, 5 and 1 h before the test): sertraline at doses of 5 and 10 mg/kg ip, pramipexole at doses of 0.05, 0.1 and 0.3 mg/kg sc. Sertraline given separately was inactive in the test used. Pramipexole reduced the immobility time only at a dose of 0.3 mg/kg. Joint administation of both those drugs distinctly shorted the immobility time, that effect.being particularly strong at pramipexole, 0.3 mg/kg, and sertraline, 5 or 10 mg/kg. The obtained results indicate that sertraline--like the previously tested citalopram and fluoxetine--shows a synergistic effect when given with pramipexole in the forced swimming test. PMID- 10817525 TI - Effect of sigma ligands on the cocaine-induced convulsions in mice. AB - It has been hypothesized that some of negative effects exerted by cocaine are mediated via sigma (sigma) receptors. This report demonstrates the effects of selective sigma ligands, panamesine, DTG, rimcazole and SA 4503, on the cocaine induced convulsions in mice and locomotor hyperactivity in rats. Only panamesine decreased both these effects of cocaine, whereas DTG and rimcazole increased the total time of cocaine-evoked convulsions and locomotor activity. SA 4503 slightly enhanced and prolonged cocaine-induced convulsions but it was ineffective in locomotor hyperactivity test. Moreover, the increase in cocaine-induced locomotor hyperactivity evoked by DTG was antagonized by panamesine. The obtained results indicate that panamesine, a selective sigma ligand with a preference for sigma1 receptor subtype and potential antagonistic activity, decreased the effects of cocaine. DTG and rimcazole (potential sigma1/sigma2 sites agonists) as well as SA 4503 (potential sigma1 site agonist) showed rather opposite effects. These findings support the idea that sigma2 receptor subtype is involved in psychomotor stimulant effects of cocaine while sigma1 receptor subtype participates in the cocaine-induced convulsions. In addition, sigma receptor antagonists (especially sigma1 ones) are able to antagonize toxic effects of cocaine while sigma agonists facilitate them. PMID- 10817526 TI - Influence of piracetam and oxiracetam on the content of high-energy phosphates and morphometry of astrocytes in vitro. AB - The nootropic drugs, including piracetam (PIR) and oxiracetam (OXI) are used in the adjunctive treatment of dementia. They are thought to directly influence energetic processes in the brain and, therefore, they are supposed to improve memory and cognition. The content of adenosine triphosphate (ATP) and phosphocreatine (PCr) and 3H-valine incorporation into proteins were measured, and the morphometry was performed after PIR and OXI treatment of astrocytes cultured in vitro with or without dibutyryl 3',5'-cyclic adenosine monophosphate (dBcAMP). Nootropics were added into the culture medium for 2 weeks at a final concentration of 10(-7) M. It was shown that OXI increased ATP content in astrocytes cultured with or without dBcAMP. The increase in 3H-valine incorporation into astrocytes after PIR and OXI together with dBcAMP treatment was found. These results indicate that the presented research model allows to study energetic processes in cultured astrocytes. However, nootropic drugs changed morphometric parameters (cell area, perimeter and form factor) of cultured astrocytes as well. It can be concluded that PIR and OXI as nootropics have an opposing effect on the content of high-energy phosphates and shape of astrocytes in vitro. PMID- 10817527 TI - Moclobemide enhances aversively motivated learning and memory in rats. AB - Moclobemide belongs to a new class of short-acting reversible monoamine oxidase type A inhibitors with antidepressant activity. Male Wistar rats (180-220 g) were administered single and multiple (14x, daily) intragastrical doses of 20 mg/kg of moclobemide. The effects of the drug on the acquisition of conditioned avoidance responses (CARs), retrieval of passive avoidance, recognition memory, psychomotor activity in open field, and level of anxiety in an elevated "plus" maze were assessed. Moclobemide, given repeatedly, increased rate of the CAR acquisition, retrieval of the passive avoidance and had some anxiolytic properties. The drug, however, did not influence rats' psychomotor activity tested in open field and their ability to discriminate between familiar and unfamiliar objects. The results show that moclobemide in rats, given chronically, enhances aversively motivated learning and does not influence recognition. PMID- 10817528 TI - Effects of intravenous anesthetic agents on pregnant myometrium. AB - Emergent or elective surgical procedures under general anesthesia may be required during pregnancy for some patients. For this reason, effects of anesthetics on uterine contractions are important. General anesthesia is not limited to the administration of inhalation agents, since induction and maintenance of anesthesia usually involve intravenous administration of different anesthetic agents. The effects of these agents on uterine contractions and blood flow are very important. The unexpected relaxation or contraction of myometrium can be harmful to fetus and continuing pregnancy. Our aim in this study is to investigate the effects of intravenous anesthetic agents on uterine contractions. In our study, we examined the effects of ketamine, propofol and midazolam on contractions of isolated pregnant rat myometrium. We observed that exposure to propofol, ketamine and midazolam at the concentrations of 10(-5)-10(-4) M decreased spontaneous contractile activity in myometrial strips isolated from pregnant rats. There was statistically significant difference between the control and all strips exposed to propofol, ketamine and midazolam in the contraction amplitude and integrated area under the contraction curve. PMID- 10817529 TI - Influence of alpha-escin on the femoral bone strength in ovariectomized rats. AB - The aim of the present study was to investigate the effect of alpha-escin (35 mg/kg po, daily) administered for 4 weeks on the femoral bone strength in 3-month old ovariectomized Wistar rats. The experiments were carried out on four groups of animals: I (C)-control sham operated rats, II (OVX)-ovariectomized rats, III (E)-sham operated rats which were administered alpha-escin, IV (OVX+E) ovariectomized rats which were administered alpha-escin. Bilateral ovariectomy caused osteopenic skeletal changes in mature female rats. alpha-Escin (35 mg/kg po, daily) administered to the ovariectomized rats for 28 days only to little extent decreased the development of osteopenic skeletal changes which were caused by bilateral ovariectomy. alpha-Escin (35 mg/kg po, daily) administered to the sham operated rats for 28 days caused slight changes in the skeletal system, which were characterized by the increase in the bone formation processes. PMID- 10817530 TI - QSAR studies on antimalarial activities of 2-phenyl-4-quinoline-carbinolamines. AB - QSAR study on a series of 2-phenyl-4-quinoline carbinolamine antimalarials was performed using electronic parameters to correlate the electronic features with antimalarial activities. The electronic parameters used were energy of the highest occupied molecular orbitals (EH), energy of the lowest unoccupied molecular orbitals (EL), the total pi-electron energy (ET) and charge density (CD). The results have shown that the antimalarial potential can be modeled by these parameters. The study will aid in efficient designing of this class of antimalarial agents. PMID- 10817531 TI - Haloperidol treatment selectively affects expression of Galpha(i)3 subunit mRNA in specific regions of the rat brain. AB - Relative abundance of GTP binding protein mRNA coding for subunits alpha(i)1, alpha(i)2 and alpha(i)3, and their changes induced with chronic blockade of D2 receptor by haloperidol were compared in the frontal cortex, striatum, hippocampus and cerebellar cortex of the rat using a specific method based on multiplex RT-PCR. Galpha(i) subtype 2 mRNA was the most abundant, followed by subtype 1 and subtype 3. The haloperidol treatment produced an area specific increase in Galpha(i)3 mRNA in the frontal cortex and a decrease of Galpha(i)3 in the striatum. After 8-day withdrawal period, haloperidol-induced,changes disappeared in the striatum but became accentuated in the frontal cortex. Changes in the relative expression of Galpha(i) subtypes may significantly influence long lasting effects of chronic D2 receptor blockade. PMID- 10817532 TI - Allopregnanolone attenuates kainate-induced toxicity in primary cortical neurons and PC12 neuronal cells. AB - In this study, we demonstrated that allopregnanolone at 500 and 1000 nM significantly inhibited kainate-induced lactate dehydrogenase release from primary cortical cells by ca. 25 and 50%, respectively. Furthermore, allopregnanolone doses of 100 and 500 nM decreased cytotoxic effects of kainate (150 microM, 24-hour exposure) in PC12 neuronal cells by about 55 and 37%, respectively. These data strongly support neuroprotective effects of allopregnanolone observed in vivo. PMID- 10817533 TI - Is serum copper a "trait marker" of unipolar depression? A preliminary clinical study. AB - Depression is not a homogenous illness and is diagnosed in only one in three persons suffering from this disorder. Besides psychological-psychiatric diagnostic methods (e.g. Hamilton's, Beck's rating scales), some biochemical measures have been introduced as markers of depression. However, the sensitivity of even the best characterized dexamethasone suppression test is as low as 40 50%. A search for better markers is continuously in progress. In the present study, we investigated alterations in serum copper concentrations in depressive patients before, during and after antidepressant treatment and compared these values with the concentrations in healthy volunteers. The serum copper concentration in depression is significantly higher (by 21%) than in the control. However, effective antidepressant treatments (which reduced symptoms by ca. 50% measured by Hamilton Depression Rating Scale) did not affect (did not normalize) this increased copper concentration. The present data indicate that serum copper is a "trait marker" (remains constant regardless of successful treatment) of unipolar depression. PMID- 10817534 TI - Effect of chronic antidepressant or electroconvulsive shock treatment on mGLuR1a immunoreactivity expression in the rat hippocampus. AB - The effect of chronic imipramine (IMI) or electroconvulsive shock (ECS) treatment on the expression of group Ia metabotropic glutamate receptors (mGluR1a) was studied in the rat hippocampus by an immunohistochemical method, using a specific monoclonal antibody. It was found that both those treatments increased the number of mGluR1a immunoreactive neurons in a pyramidal layer of the CA3 hippocampal field. Moreover, IMI, but not ECS, increased the density of mGluR1 a positive neurons in the hilus. The obtained results indicate a possible influence of the antidepressive treatment on the mGluR1a expression in some hippocampal fields. PMID- 10817535 TI - Selective group II glutamate metabotropic receptor agonist LY354740 attenuates pentetrazole- and picrotoxin-induced seizures. AB - There are several data indicating the involvement of metabotropic glutamate receptors (mGluR) in seizures and epileptogenesis. In the present experiments, the selective group II mGluR agonist (+)-2-aminobicyclo-[3.1.0]hexane-2,6 dicarboxylic acid (LY 354740) at doses from 4 to 16 mg/kg) administered prior to the injection of pentetrazole (80 mg/kg) or picrotoxin (3.2 mg/kg) produced a dose-dependent decrease in the number of mice exhibiting clonic convulsions. Our data suggest that group II mGluR agonist LY 354740 possesses antiseizure activity. PMID- 10817536 TI - Leukocyte subsets in treatment-resistant major depression. AB - Patients with treatment-resistant major depression had a significantly higher percentage of helper CD4+ T cells and immature double positive CD4+CD8+ T cells in their blood, and higher CD4+/CD8+ ratio than healthy controls. These results suggest activation of T helper immuno-response and disturbance in thymus function in these patients. PMID- 10817538 TI - Lack of effect of a glycine(B) receptor partial agonist on amphetamine-induced sensitization in mice. AB - Effects of 1-aminocyclopropanecarboxylic acid (ACPC), a partial agonist of glycine(B) receptors, on the expression and development of sensitization to the locomotor activity of amphetamine (AMPH, 2.5 mg/kg) were studied in mice. ACPC in doses of 100-400 mg/kg did not affect the expression of AMPH sensitization. Combined injections of ACPC (200-400 mg/kg) and AMPH during the development of sensitization did not change the expression of sensitization to the challenge dose of AMPH 3 days after the drug withdrawal. Acute administration of ACPC, 400 mg/kg, enhanced the locomotor hyperactivity induced by a single dose of AMPH, that effect being probably connected with its own stimulatory action of that dose of ACPC. Summing up, our results show that ACPC affects neither the development nor the expression of AMPH sensitization in mice. PMID- 10817537 TI - Adrenergic receptors' responsiveness after acute and chronic treatment with haloperidol in the presence of calcium channel blockade. AB - The effects of acute and chronic (14 days) treatment with haloperidol (0.5 mg/kg ip) given alone or 15 min after nifedipine (5 mg/kg ip) on responsiveness of the alpha1- and beta-adrenergic receptors was investigated ex vivo by measuring of the second messenger responses. The accumulation of inositol phosphate (IP) or cyclic AMP were measured in brain cortical slices challenged with noradrenaline or isoproterenol. The facilitatory effect of a single dose of haloperidol on IP accumulation (an increase by approximately 50%) was abolished by nifedipine pretreatment. After chronic treatment with haloperidol alone its facilitatory effect disappeared, but a treatment with haloperidol after nifedipine caused a significant increase in IP accumulation. No treatment affected the responses from beta-adrenoceptors. These data show that the action of chronically administered haloperidol may be changed by the concomitant calcium channel blockade that prevents development of adaptation to persistent presence of the drug. PMID- 10817539 TI - Reversal of stress-induced deficit in aggression by monoamine oxidase inhibitors. AB - In the present study we investigated the effect of two monoamine oxidase (MAO) inhibitors: moclobemide (selective, reversible inhibitor of MAO-type A) or selegiline (selective irreversible inhibitor of MAO-type B) on electric footshock induced fighting behavior in normal (unstressed) and chronically stressed (14 various stressors over 16 days) rats. In rats exposed to chronic stress the number of fighting attacks was reduced by about 75%. Prolonged (once a day, for 14 days) treatment with moclobemide (50 mg/kg/day) or selegiline (2 mg/kg/day) counteracted the deficit in aggression induced by chronic stress. The findings of the present study demonstrate that the selective MAO inhibitors, moclobemide and selegiline, protect against "behavioral depression" induced by the chronic stress similarly to other classes of antidepressant drugs. PMID- 10817540 TI - Pharmacological profile of reboxetine, a representative of new class of antidepressant drugs, selective noradrenaline reuptake inhibitor (NARI), given acutely. AB - Pharmacological effects of acute treatment with reboxetine (REB), a clinically active antidepressant (a noradrenaline reuptake inhibitor without any affinity for neurotransmitter receptors), were studied in mice and rats. REB inhibited the reserpine- or apomorphine-induced hypothermia in mice. It reduced the immobility time in Porsolt's test in mice and rats, but it did not change the locomotor activity in mice and rats. REB changed neither the clonidine-induced aggressiveness in mice nor the behavioral syndrome induced by oxotremorine in rats. The obtained results indicate that REB, given acutely, shows a pharmacological profile similar to that of tricyclic or tetracyclic noradrenaline reuptake inhibitors. In contrast to the antidepressants mentioned above, REB does not exhibit an alpha1-adrenolytic or cholinolytic activity (in vivo tests). PMID- 10817541 TI - Some pharmacological properties of new analogs of MP 3022, the 5-HT1A receptor antagonist. AB - Two new analogs of full 5-HT1A receptor antagonist 4-[3-(1-benzotriazolyl)propyl] 1-(2-methoxyphenyl)piperazine (MP 3022; 1) containing di- (5) or tetramethylene- (6) spacer were synthesized. In the radioligand binding studies, compounds 5 and 6 showed high 5-HT1A (Ki = 14.7 nM and 11.8 nM, respectively) and low 5-HT2 receptor affinity (Ki = 2,696 nM and 389.2 nM, respectively). In behavioral studies both compounds behaved like postsynaptic 5-HT1A receptor antagonists as they reduced lower lip retraction and behavioral syndrome induced by 8-OH-DPAT (5 HT1A receptor agonist) in rats, but 6 was more effective in these tests. Derivative 5 did not affect body temperature in mice, whereas 6 decreased it. Furthermore, 5 did not change hypothermia induced by 8-OH-DPAT, and 6-induced lowering of body temperature in mice was not antagonized by (S)-WAY 100135 (5 HT1A antagonist), so in that model 5 and 6 did not behave as antagonist or agonist, respectively, at presynaptic 5-HT1A receptors. Compound 6 was studied in behavioral tests used to predict a potential anxiolytic (conflict drinking test in rats) and antidepressant (forced swimming test in rats) activity. Diazepam and imipramine were used as reference drugs. Compound 6 significantly increased the number of shocks accepted in water-deprived rats in conflict drinking test and shortened the immobility time in forced swimming test in rats. The above findings indicate that new 5-HT1A postsynaptic antagonist 6 behaves like anxiolytic and antidepressant, but mechanisms of these properties of 6 remain unknown. PMID- 10817542 TI - Effect of the amide fragment on 5-HT1A receptor activity of some analogs of MP 3022. AB - A new set (3-11) of analogs of MP 3022 (1) containing the amide bond inserted into the intermediate chain linking the terminal heteroaromatic and 1-(2 methoxyphenyl)piperazine moieties were prepared and their 5-HT1A and 5-HT2A receptor affinities were determined. Only compounds with trimethylene chain between amide and arylpiperazine fragments (5a, 5b, 7a, 7b and 11) showed satisfactory affinity for 5-HT1A receptor (Ki = 42-87 nM) and high 5-HT2A/5-HT1A selectivity. The new 5-HT1A receptor ligands were investigated in vivo to determine their 5-HT1A agonistic or antagonistic properties. Compounds 7a and 7b with terminal indazole fragment as well as 11 with phenyl substituent behaved like weak 5-HT1A receptor antagonists. The structure-affinity relationship studies in this series of compounds revealed that the amide group along with the terminal aromatic fragments contributed to interaction with 5-HT1A receptor sites, whereas in vivo results indicated that introduction of the amide group into presented arylpiperazine structures was not a profitable modification for their 5-HT1A functional activity. PMID- 10817543 TI - Imipramine-induced increase in the inhibitory effect of adenosine receptor activation in the hippocampus. AB - Imipramine, a tricyclic antidepressant, is one of the main drugs used for the treatment of depression. We investigated the effects of the repeated administration of imipramine (10 mg/kg po for 14 days, twice daily) on adenosine receptor-mediated actions using extracellular and intracellular recording techniques in the rat hippocampal slices. Adenosine and 2-chloroadenosine dose dependently decreased the amplitude of population spikes and the slope of the field excitatory postsynaptic potentials (fEPSPs) evoked in the CA1 cell layer and apical dendrites of the CA1 cells, respectively, by stimulation of the Schaffer collateral/commissural pathway. As revealed by intracellular recording, a membrane hyperpolarization and a strong attenuation of excitatory synaptic transmission contribute to the decrease in the population spikes and fEPSPs induced by adenosine and 2-chloroadenosine. The repeated administration of imipramine enhanced the effect of adenosine (3 microM) and 2-chloroadenosine (0.15 microM) on fEPSPs while the inhibition of population spikes was not changed. When higher concentration of 2-chloroadenosine (0.25 microM) was tested, repeated imipramine administration enhanced its inhibitory effect on population spikes but not on fEPSPs. The present report provides evidence that the inhibitory effect of adenosine receptor activation in the hippocampus is enhanced by repeated treatment with imipramine. PMID- 10817544 TI - Protective effect of alpha-keto acids on the oxidative hemolysis. AB - We studied antioxidative properties of various concentrations (0.3-2 mM) of biochemically important alpha-keto acids: pyruvate, alpha-ketoglutarate, oxaloacetate, glyoxylic acid as aldehyde acid and also 2-methyl-thiazolidyne-2,4 dicarboxylic acid and their effect on the oxidative hemolysis of human erythrocytes induced by hydrogen peroxide (H2O2). Normal erythrocytes proved to be very resistant to oxidative damages, so the high concentration of H2O2 (10 mM) as well as the presence of natrium azide, a catalase inhibitor, was necessary. The levels of malonyldialdehyde (MDA), reactive oxygen species (ROS) and hemoglobin (Hb) released were evaluated as the measure of red cell peroxidative hemolysis. Pyruvate, at the lowest used concentration (0.3 mM), caused the inhibition of lipid peroxidation (MDA) and a drop in the level of ROS, as well as a diminution of the degree of hemolysis and the effects were stronger than those of other alpha-keto acids. At the highest (2 mM) concentration, the protective effect against oxidative damage of all the investigated alpha-keto acids was similar and amounted to nearly 50% in relation to the control sample. On the contrary, in the case of aldehyde acid, e.g. glyoxylic acid, no protective effect in the same range of concentrations was found. This confirms the participation of non-enzymatic oxidative decarboxylation of alpha-keto acids in the hydrogen peroxide decomposition process. PMID- 10817545 TI - Effects of selective cytochrome P-450 inhibitors on the metabolism of thioridazine. In vitro studies. AB - The aim of the present study was to determine optimum conditions for the study of thioridazine metabolism in rat liver microsomes and to investigate the influence of specific cytochrome P-450 inhibitors on 2- and 5-sulfoxidation, and N demethylation of thioridazine. Basing on the developed method, the thioridazine metabolism in liver microsomes was studied at linear dependence of the product formation on time, and protein and substrate concentrations (incubation time was 15 min, concentration of microsomal protein was 0.5 mg/ml, substrate concentrations were 25, 50 and 75 nmol/ml). Dixon analysis of tioridazine metabolism carried out in the control liver microsomes, in the absence and presence of specific cytochrome P-450 inhibitors, showed that quinine (CYP2D1 inhibitor), metyrapone (CYP2B1/B2 inhibitor) and alpha-naphthoflavone (CYP1A2 inhibitor) affected while erythromycin (CYP3A inhibitor) and sulfaphenazole (CYP2C9 inhibitor) did not affect the neuroleptic biotransformation. Thus, quinine and metyrapone inhibited competitively thioridazine N-demethylation and mono-2-sulfoxidation. As reflected by Ki values, N-demethylation was inhibited to a higher degree (Ki = 16.5 and 43 microM, respectively) than mono-2-sulfoxidation (Ki = 25 and 137 microM, respectively). On the other hand, alpha-naphthoflavone inhibited competitively not only N-demethylation and mono-2-sulfoxidation, but also 5-sulfoxidation of thioridazine. The calculated Ki values showed that the highest potency of alpha-naphthoflavone to inhibit thioridazine metabolism was observed for N-demethylation and it descended in the following order: N demethylation (Ki = 13.8 microM) > mono-2-sulfoxidation (Ki = 34 microM) > 5 sulfoxidation (Ki = 70.4 microM). In conclusion, it can be assumed that N demethylation and mono-2-sulfoxidation are catalyzed by the isoenzymes 2D1, 2B and 1A2 while 5-sulfoxidation only by 1A2; isoenzymes belonging to the subfamilies 2C and 3A seem not to be involved in the metabolism of thioridazine. The obtained results are discussed in the view of species and structure differences in the enzymatic catalysis of phenothiazines' metabolism as well as in relation to their pharmacological and clinical significance. PMID- 10817546 TI - L-N6-(1-iminoethyl)-lysine (L-NIL) but not S-methylisothiourea sulphate (SMT) displays selectivity towards NOS-2. AB - Our aim was to verify potency and selectiveness of two most widely used drugs regarded as NOS-2 inhibitors: L-N6-(1-iminoethyl)-lysine (L-NIL) and S methylisothiourea sulphate (SMT). Thioglycolate-elicited rat peritoneal macrophages and coronary endothelium of isolated guinea pig heart were used as assay systems for NOS-2 and NOS-3, respectively. A non-selective NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME) was used as a reference compound. We found that L-NIL but not SMT was a selective NOS-2 inhibitor. Interestingly, L NAME displayed selectivity towards NOS-3. PMID- 10817547 TI - Molecular basis and clinical use of biochemical markers of bone, cartilage, and synovium in joint diseases. PMID- 10817548 TI - Sonographic imaging of tendons. PMID- 10817549 TI - Rofecoxib, a specific inhibitor of cyclooxygenase 2, with clinical efficacy comparable with that of diclofenac sodium: results of a one-year, randomized, clinical trial in patients with osteoarthritis of the knee and hip. Rofecoxib Phase III Protocol 035 Study Group. AB - OBJECTIVE: To compare the clinical efficacy of rofecoxib, a specific inhibitor of cyclooxygenase 2 (COX-2), with that of diclofenac in patients with osteoarthritis (OA) and to evaluate the safety and tolerability of rofecoxib. METHODS: We performed a randomized, double-blind, active comparator-controlled trial in 784 adults with OA of the knee or hip. Patients were randomized to 1 of 3 treatment groups: 12.5 mg of rofecoxib once daily, 25 mg of rofecoxib once daily, and 50 mg of diclofenac 3 times daily. Clinical efficacy and safety were evaluated over a 1 year continuous treatment period. RESULTS: Rofecoxib at dosages of 12.5 and 25 mg demonstrated efficacy that was clinically comparable to that of diclofenac, as assessed by all 3 primary end points according to predefined comparability criteria. Results from secondary end points were consistent with those of the primary end points. There were small statistical differences favoring diclofenac for 2 of the end points. All treatments were well tolerated. CONCLUSION: Rofecoxib was well tolerated and provided efficacy that was clinically comparable, according to predefined statistical criteria, to that of 150 mg of diclofenac per day in this 1-year study. Specific inhibition of COX-2 provided therapeutic efficacy in OA. PMID- 10817550 TI - Quantification of progressive joint space narrowing in osteoarthritis of the hip: longitudinal analysis of the contralateral hip after total hip arthroplasty. AB - OBJECTIVE: The rate of progressive joint space narrowing in the contralateral hip after total hip arthroplasty (THA) for osteoarthritis (OA) and the factors which may predispose patients to more aggressive joint space narrowing remain undefined. The current study sought to evaluate the rate and pattern of, and risk factors for, progressive joint space narrowing in the contralateral hip after THA for OA. METHODS: Each patient who underwent THA for OA in 1984-1985 was followed up longitudinally, and annual anteroposterior (AP) pelvis radiographs were obtained. The radiographic joint space width (JSW) of each contralateral hip joint was quantified, and the rates of JSW narrowing were determined. Evaluation of potential risk factors for accelerated progression of joint space narrowing included age, sex, side of surgery, weight, height, body mass index (BMI), hip pain, etiology of OA, and Kellgren/Lawrence radiographic grade. RESULTS: Ninety nine patients and 619 AP pelvis radiographs were evaluated. The median initial JSW was 3.48 mm (interquartile range 1.55). JSW declined in a linear manner at a median rate of 0.10 mm/year. The rate of decline between baseline and followup in 20 months was predictive of the overall slope. Two subpopulations were identified. Eighty-five percent of patients maintained a slow decline in JSW (< or =0.2 mm/year), and 15% exhibited an accelerated decline in JSW (>0.2 mm/year). Kellgren/ Lawrence radiographic grade > or =2 and a diagnosis of primary OA were each associated with a more rapid decline in JSW (P = 0.006 and P = 0.02, respectively). Initial JSW, age, sex, weight, height, BMI, and hip pain were not risk factors for rapid decline in JSW. CONCLUSION: Radiographic hip JSW may be reliably quantified and followed up longitudinally using standard AP radiographs. Progression of JSW narrowing in the contralateral hip after THA for OA proceeds in a linear manner over several years. A subpopulation of patients with accelerated narrowing of contralateral JSW may be identified within 20 months, and may represent a suitable population with which to assess the potential efficacy of new disease-modifying agents. PMID- 10817551 TI - Risk factors for the incidence and progression of radiographic knee osteoarthritis. AB - OBJECTIVE: Preventive strategies against knee osteoarthritis (OA) require a knowledge of risk factors that influence the initiation of the disorder and its subsequent progression. This population-based longitudinal study was performed to address this issue. METHODS: Ninety-nine men and 255 women aged > or =55 years had baseline interviews and weight-bearing knee radiographs in 1990-1991. Repeat radiographs were obtained in 1995-1996 (mean followup duration 5.1 years, median age at followup 75.8 years). Risk factors assessed at baseline were tested for their association with incident and progressive radiographic knee OA by logistic regression. RESULTS: Rates of incidence and progression were 2.5% and 3.6% per year, respectively. After adjusting for age and sex, the risk of incident radiographic knee OA was significantly increased among subjects with higher baseline body mass index (odds ratio [OR] 18.3, 95% confidence interval [95% CI] 5.1-65.1, highest versus lowest third), previous knee injury (OR 4.8, 95% CI 1.0 24.1), and a history of regular sports participation (OR 3.2, 95% CI 1.1-9.1). Knee pain at baseline (OR 2.4, 95% CI 0.7-8.0) and Heberden's nodes (OR 2.0, 95% CI 0.7-5.7) were weakly associated with progression. Analyses based on individual radiographic features (osteophyte formation and joint space narrowing) supported differences in risk factors for either feature. CONCLUSION: Most currently recognized risk factors for prevalent knee OA (obesity, knee injury, and physical activity) influence incidence more than radiographic progression. Furthermore, these factors might selectively influence osteophyte formation more than joint space narrowing. These findings are consistent with knee OA being initiated by joint injury, but with progression being a consequence of impaired intrinsic repair capacity. PMID- 10817553 TI - Breast-feeding and postpartum relapse in women with rheumatoid and inflammatory arthritis. AB - OBJECTIVE: To test the hypothesis that breast-feeding increases the risk of postpartum flare in inflammatory polyarthritis. METHODS: We compared disease activity during pregnancy and at 6 months postpartum among 49 non-breast-feeders, 38 first-time breast-feeders, and 50 repeat breast-feeders. RESULTS: After we adjusted for possible confounders, including treatment, first-time breast-feeders had increased disease activity 6 months postpartum, based on self-reported symptoms, joint counts, and C-reactive protein levels. CONCLUSION: Postpartum flare may be induced by breast-feeding. PMID- 10817552 TI - A multicenter, double-blind, dose-ranging, randomized, placebo-controlled study of recombinant human interleukin-1 receptor antagonist in patients with rheumatoid arthritis: radiologic progression and correlation of Genant and Larsen scores. AB - OBJECTIVE: To evaluate radiographic progression and the relationship of radiologic scores obtained by the Genant and Larsen methods in a clinical trial of recombinant human interleukin-1 receptor antagonist (IL-1Ra). METHODS: Patients with rheumatoid arthritis (RA) were randomized into 4 groups: placebo (n = 121) or IL-1Ra at a daily dosage of 30 mg (n = 119), 75 mg (n = 116), or 150 mg (n = 116). Hand radiographs obtained at baseline, 24 weeks, and 48 weeks were scored using both methods. RESULTS: At 24 weeks, by the Genant method, there was significant reduction in the score for progression of joint space narrowing (JSN) and the total score (a combination of erosion and JSN) in all treatment groups. Least-squares mean changes in the Genant erosion score from baseline to 24 weeks were significantly reduced after treatment with IL-1Ra at 30 mg/day and for all IL-1Ra treatment groups combined. The changes corresponded to a reduction of 38% in erosion, 58% in JSN, and 47% in total score. Patients treated with IL-1Ra at 75 mg/day had a significant reduction in the Larsen erosive joint count (LEJC), and all IL-1RA-treated groups combined showed a 45% reduction. Correlations (r) between the Genant total and Larsen scores were 0.84 at baseline, 0.83 at week 24, and 0.83 at week 48 (P < 0.0001); correlations between the Genant erosion score and the LEJC were 0.83 (P < 0.0001) at all visits; correlations between the Genant total and the Larsen scores were 0.32 and 0.49 (P < 0.0001) for progression from baseline to week 24 and from baseline to week 48, respectively; correlations between the Genant erosion score and the LEJC were 0.36 and 0.41 (P < 0.0001) for progression to weeks 24 and 48, respectively. CONCLUSION: IL-1Ra reduced radiologic progression of RA. Scores by the 2 methods correlated strongly for each individual time point, but much less strongly for assessments of disease progression. PMID- 10817554 TI - Single values of serum transferrin receptor and transferrin receptor ferritin index can be used to detect true and functional iron deficiency in rheumatoid arthritis patients with anemia. AB - OBJECTIVE: To elucidate the use of serum transferrin receptor (sTfR) to distinguish between iron-deficiency anemia (IDA) and anemia of chronic disease (ACD), and to establish an improved scheme to identify functional iron deficiency (FID) in rheumatoid arthritis (RA) patients with anemia. METHODS: We studied 30 anemic RA patients whose iron status was confirmed by bone marrow examination and determination of the sTfR level, serum ferritin level, and sTfR-log ferritin index (TfR-F Index). All patients with diminished or exhausted iron stores (n = 18) received oral iron supplementation. RESULTS: Baseline values of sTfR and the TfR-F Index predicted the response correctly in all patients who received supplementation treatment and were normal in 10 of 11 patients with normal initial iron stores (ACD). CONCLUSION: The results of this study elucidate the roles of sTfR and the TfR-F Index in the differential diagnosis between IDA and ACD and provide direct evidence that these parameters are useful in detecting FID, irrespective of the concurrent iron storage status. PMID- 10817555 TI - An interdisciplinary approach to the care of patients with Wegener's granulomatosis: long-term outcome in 155 patients. AB - OBJECTIVE: To examine the outcome in 155 consecutive patients with Wegener's granulomatosis (WG) followed up for a median of 7 years. METHODS: Treatment was adapted to the activity and extent of disease, with regular evaluation by an interdisciplinary team accompanied by group education about vasculitis. RESULTS: The estimated median survival time was 21.7 years (95% confidence interval [95% CI] 15.60-27.86). Twenty-two patients died; 19 deaths were attributable to WG and/or its treatment. Significant predictors of survival at diagnosis were age >50 years (hazard ratio [HR] 5.45, 95% CI 1.97-15.02), kidney involvement with impaired renal function (HR 5.42, 95% CI 1.76-16.68), and lung involvement (HR 3.75, 95% CI 1.26-11.16). At some stage, 142 patients received prednisone and cyclophosphamide (CYC), usually as daily CYC plus mesna as uroprotection, 50 patients received trimethoprim/sulfamethoxazole, and 45 received methotrexate. Complete remission was achieved in 83 of the 155 patients. One or more relapses occurred in 99 patients after either complete or partial remission. CYC-induced cystitis and myelodysplastic syndrome occurred in 17 and 11 patients, respectively. A cumulative dose of 100 gm or more of CYC resulted in a 2-fold greater risk of CYC-related morbidity than with lower CYC doses. Serious infections occurred in 41 patients. CONCLUSION: An interdisciplinary approach to the care of 155 WG patients resulted in a median survival of >21 years. Kidney or lung involvement at diagnosis was predictive of a >3-fold higher mortality. Although CYC remains essential in the treatment of WG, it was administered as briefly as possible and under close surveillance to avoid permanent CYC-related morbidity, which can lead to serious therapeutic problems in chronic relapsing WG. PMID- 10817556 TI - Chylomicron metabolism is markedly altered in systemic lupus erythematosus. AB - OBJECTIVE: To verify the in vivo status of chylomicron metabolism in systemic lupus erythematosus (SLE) since there is a high incidence of atherosclerosis in this disease and chylomicrons may have an important role in atherogenesis. METHODS: A chylomicron-like emulsion labeled with 14C-cholesteryl esters and 3H triglycerides was injected intravenously into 10 female patients with inactive SLE and 10 healthy age- and sex-matched control subjects to determine the plasma kinetics of the emulsion lipids from consecutive plasma samples taken at regular intervals for 1 hour. Lipolytic activity was determined in vitro after incubation of the labeled emulsion with postheparin plasma. RESULTS: The decay curves for the emulsion were markedly slowed in SLE. Chylomicron lipolysis, indicated by the fractional clearance rate (FCR) of emulsion 3H-triglyceride, was 2-fold smaller in SLE patients than in controls (mean +/- SD 0.023 +/- 0.011 versus 0.047 +/ 0.015 minute(-1); P = 0.010). Chylomicron removal, indicated by emulsion 14C cholesteryl ester FCR, was 3-fold smaller in SLE patients than in controls (0.007 +/-0.007 versus 0.023 +/- 0.011 minute(-1); P = 0.009). In vitro lipolysis in SLE patients was nearly half that of the controls (mean +/- SD 10,199 +/- 2,959 versus 6,598 +/-2,215; P = 0.014). Higher levels of very-low-density lipoprotein cholesterol and triglycerides and lower levels of high-density lipoprotein cholesterol and apolipoprotein A-I were also observed in the SLE patients. CONCLUSION: SLE patients have disturbances in chylomicron metabolism that are characterized by decreased lipolysis and chylomicron remnant removal from the plasma. This finding, together with other alterations in lipid profiles that were confirmed in the present study, is largely accountable for the accelerated atherosclerotic process of the disease. PMID- 10817557 TI - Treatment of giant cell arteritis: interleukin-6 as a biologic marker of disease activity. AB - OBJECTIVE: To determine the value of the erythrocyte sedimentation rate (ESR) and plasma interleukin-6 (IL-6) as biologic markers for monitoring disease activity in giant cell arteritis (GCA). METHODS: Twenty-five patients with biopsy-proven GCA were enrolled into a prospective treatment study. Therapy was initiated with prednisone, 60 mg/day, followed by a predetermined tapering schedule. Patients were monitored monthly for clinical signs of active vasculitis and laboratory parameters indicative of inflammation, including elevated ESR (>30 mm/hour) and elevated plasma IL-6 concentrations (>6.1 pg/ml). RESULTS: Upon initiation of corticosteroid treatment, clinical signs of GCA disappeared in all patients; however, 60% of the patients developed symptoms of recurrent disease, on 1 or more occasions, while the prednisone dosage was being reduced. These 31 disease flares diagnosed over 550 days were associated with symptoms of systemic inflammation but did not result in vascular complications. The ESR was elevated in 76% of the patients prior to initiation of treatment (median 65 mm/hour) and normalized by day 28 of therapy (median 6 mm/hour). The median ESR remained in the normal range during the followup period. Plasma IL-6 levels, which were abnormal in 92% of untreated patients (median 16 pg/ml), were partially responsive to the initial high doses of corticosteroids by day 28 (median 6 pg/ml), but levels did not completely normalize with continued therapy. Elevation of the ESR was seen during only 58% of all disease flares, but 89% of disease recurrences were associated with increased plasma IL-6 levels (P = 0.03). CONCLUSION: Plasma IL-6 is more sensitive than ESR for indicating disease activity in untreated and treated GCA patients. Standard corticosteroid regimens only partially suppress vascular inflammation. Smoldering disease activity may expose GCA patients to the risk of progressive vascular disease (e.g., formation of aortic aneurysms) and chronic systemic complications such as IL-6-mediated osteopenia. PMID- 10817558 TI - QT interval prolongation in asymptomatic anti-SSA/Ro-positive infants without congenital heart block. AB - OBJECTIVE: To analyze the electrocardiograms (EKGs) of infants born to mothers with anti-SSA/Ro antibodies in order to evaluate the QT interval (the time from the beginning of the QRS complex to the end of the T wave). METHODS: Sera from mothers and children were analyzed for anti-Ro and anti-SSB/La antibodies by enzyme-linked immunosorbent assay (ELISA) and by Western blot analysis. Fine specificity of anti-Ro antibodies was evaluated by solid-phase ELISA against recombinant 52- and 60-kd proteins and by Western blot. A retrospective chart review was conducted for EKG analysis. Twenty-eight EKG tracings (21 from anti-Ro positive and 7 from anti-Ro-negative infants born to mothers with autoimmune diseases) were analyzed by a single investigator who was blinded to the infant's antibody status. The QT interval was measured and corrected for heart rate according to Bazett's formula. RESULTS: The mean QT interval was significantly longer in anti-Ro-positive than in anti-Ro-negative infants, also after correction for heart rate (QTc) (P = 0.001). Nine of 21 anti-Ro-positive infants and 0 of 7 anti-Ro-negative infants had QTc values above the upper normal limit (440 msec). A 24-hour EKG recording was performed on 5 patients and confirmed the QT prolongation. These infants were subsequently treated with a beta-blocker in order to prevent arrhythmias. CONCLUSION: Infants born to mothers who carry anti Ro autoantibodies may show QT interval prolongation and should be monitored with EKG during the first months of life. PMID- 10817559 TI - Induction of immune tolerance to human type I collagen in patients with systemic sclerosis by oral administration of bovine type I collagen. AB - OBJECTIVE: To determine whether oral tolerance to type I collagen (CI) could be induced in patients with systemic sclerosis (SSc). METHODS: Twenty adult patients with limited or diffuse SSc were enrolled in a study to receive 0.1 mg of solubilized native bovine CI daily for 1 month, followed by 0.5 mg daily for 11 months. Peripheral blood mononuclear cells (PBMC) were obtained from the patients and cultured with human alpha1(I) and alpha2(I) chains, before and after CI treatment. Culture supernatants were analyzed for levels of interferon-gamma (IFNgamma) and interleukin-10 (IL-10). Sera obtained before and after treatment were analyzed for levels of soluble IL-2 receptor (sIL-2R). Although this study was not intended to assess the clinical efficacy of oral CI administration in SSc, selected measures of disease severity and organ involvement were evaluated. RESULTS: Oral administration of CI to SSc patients induced significant reductions in levels of IFNgamma and IL-10 in alpha1(I)- and alpha2(I)-stimulated PBMC culture supernatants, indicating that T cell immunity to CI was decreased by this treatment. Serum levels of sIL-2R also decreased significantly after oral CI treatment, suggesting a reduction in T cell activation. Significant improvements occurred in the modified Rodnan skin thickness score and the modified Health Assessment Questionnaire after 12 months of oral CI in this open trial. The lung carbon monoxide diffusing capacity improved statistically and showed a trend toward clinically significant improvement. CONCLUSION: Oral administration of bovine CI to patients with diffuse or limited SSc induces a reduction in T cell reactivity to human CI, appears to be well tolerated, and does not worsen the disease. Further evaluation of oral tolerance to CI in patients with SSc is justified to determine whether it has therapeutic efficacy. PMID- 10817560 TI - Detection of cellular microchimerism of male or female origin in systemic sclerosis patients by polymerase chain reaction analysis of HLA-Cw antigens. AB - OBJECTIVE: Systemic sclerosis (SSc; scleroderma), a disease of unknown origin, displays many similarities to chronic graft-versus-host disease. It occurs most frequently in women after the childbearing years. In recent studies, the presence of Y-chromosome DNA derived from male fetuses was detected, but DNA from female fetuses could not be investigated in those studies. The present study was undertaken to investigate cellular microchimerism of either male or female origin in DNA from patients with SSc. METHODS: DNA from peripheral blood cells of 63 patients with SSc, 64 healthy individuals, and 24 non-SSc disease controls was examined by polymerase chain reaction analysis of HLA-Cw antigens. RESULTS: Cellular microchimerism of either male or female origin was detected in 41 of 63 SSc patients (65%), in contrast to 18 of 64 normal controls (28%) (chi2 = 15.98, Pcorr = 0.00006) and 8 of 24 disease controls (33%)(chi2= 5.89, Pcorr, = 0.015). CONCLUSION: These findings support the hypothesis that microchimeric cells persisting in the circulation or tissues of SSc patients could be involved in disease pathogenesis by initiating a graft-versus-host-like reaction. PMID- 10817561 TI - Microsatellites and intragenic polymorphisms of transforming growth factor beta and platelet-derived growth factor and their receptor genes in Native Americans with systemic sclerosis (scleroderma): a preliminary analysis showing no genetic association. AB - OBJECTIVE: Abnormalities of transforming growth factor beta (TGFbeta) and platelet-derived growth factor (PDGF) alpha and beta and/or their receptors have been demonstrated in systemic sclerosis (SSc). This study aimed to determine whether genetic polymorphisms in or near the TGFbeta and PDGF gene families were associated with susceptibility to SSc in a Native American population with a high disease prevalence. METHODS: Genotyping of 5 intragenic polymorphisms within the TGFbeta1 gene and mapping of 35 microsatellites near the genes for TGFbeta1, latent TGFbeta1 binding protein (LTBP1), TGFbeta receptors I and II, PDGFalpha, PDGFbeta, PDGF receptor alpha, and PDGF receptor beta was performed in 19 SSc patients, 76 controls, and 42 family members. Allele distributions and frequencies were examined between SSc patients and controls, and marker haplotypes were examined in families when allele frequencies appeared to be different between patients and controls. RESULTS: Although 1 polymorphism within the TGFbeta1 gene (TGFbeta1) was modestly increased in the SSc patients, this did not maintain statistical significance after correction. Similarly, 1 microsatellite (D9S120) near the TGFbeta receptor I gene (TGFBR1) showed a significant disturbance of allele frequencies between patients and controls; however, it did not form a disease-associated haplotype with other nearby markers. Weak disturbances of markers near PDGFalpha (PDGFA) and PDGFbeta, (PDGFB) also failed to maintain significance after correction. Both PDGF receptor genes (PDGFRA and PDGFRB) also showed no disease associations. CONCLUSION: The results of these preliminary analyses suggest that genetic anomalies of the TGFbeta1 and PDGF gene families are not likely to explain the dysregulation seen in SSc or to account for the susceptibility to SSc in this population. PMID- 10817562 TI - Longitudinal analysis of autoantibody response to topoisomerase I in systemic sclerosis. AB - OBJECTIVE: To examine serial changes in serum anti-topoisomerase I (anti-topo I) antibody levels in patients with systemic sclerosis (SSc), as well as associations with clinical features and the in vivo activation status of circulating topo I-reactive T and B cells. METHODS: Serum anti-topo I antibody levels were serially measured at different time points in 28 SSc patients who were positive for anti-topo I antibody at their first visit (range of followup 6 29 years). The patients were subgrouped according to the disappearance (group 1) or persistence (group 2) of anti-topo I antibody. Clinical findings as well as T and B cell responses to topo I were compared between these 2 groups. RESULTS: Serum anti-topo I antibody disappeared during the period of followup in 6 patients (group 1), but persisted in 22 patients (group 2). Loss of anti-topo I antibody occurred within 10 years after the first visit and independently of treatment. Group 1 patients had less extensive skin and lung involvement and better survival rates than did group 2 patients. Complete loss of anti-topo I antibody followed a reduction in isotype expression and epitope reactivities. The kinetics of in vitro T cell proliferation induced by topo I were delayed and circulating topo I-reactive T cells were less frequently detected in group 1 versus group 2 patients, suggesting that the disappearance of anti-topo I antibody was due to loss of activation of topo I-reactive T cells. In vitro production of anti-topo I antibody in peripheral blood mononuclear cell cultures in response to antigenic stimulation in both group 1 and group 2 patients indicated persistence of anti-topo I antibody-producing "memory" B cells even after the loss of serum anti-topo I antibody. CONCLUSION: Our results indicate that there is a distinct subset of anti-topo I-positive SSc patients who lose anti-topo I antibody during the disease course and have a favorable outcome. In vivo production of anti-topo I autoantibody may require antigenic stimulation that activates topo I-reactive T and B cells. PMID- 10817563 TI - Correlation between increased nitric oxide production and markers of endothelial activation in systemic sclerosis: findings with the soluble adhesion molecules E selectin, intercellular adhesion molecule 1, and vascular cell adhesion molecule 1. AB - OBJECTIVE: To determine the relationship between vascular function and the inflammatory response in systemic sclerosis (SSc), and to investigate whether production of endothelial-derived nitric oxide (NO) is disturbed in this disease. METHODS: We measured plasma nitrate, urinary excretion of both nitrate and cGMP, and soluble adhesion molecules of endothelial origin in patients with SSc and in age- and sex-matched controls and compared these levels between groups. Additionally, we performed correlation analysis to determine how these variables were related to one another. Plasma nitrate and 24-hour-urinary excretion of nitrate in patients and controls were measured after a 72-hour nitrate-free-diet, using a gas chromatography/mass spectrometric method. Soluble adhesion molecules intercellular adhesion molecule 1 (sICAM-1), vascular cell adhesion molecule 1 (sVCAM-1), and E-selectin and cytokines were measured by enzyme-linked immunosorbent assay. The expression of E-selectin was further investigated in skin biopsy specimens by immunoperoxidase staining, and the presence of inducible NO synthase by immunoblotting. RESULTS: Plasma nitrate and 24-hour-urinary excretion of cGMP were significantly elevated in patients compared with controls, while 24-hour-urinary-excretion of nitrate tended to be elevated in SSc patients. Levels of sICAM-1, sVCAM-1, and sE-selectin were significantly elevated in the patients. Levels of plasma nitrate in the patients correlated significantly with levels of sVCAM-1 (P = 0.020) and sE-selectin (P = 0.018) and approached a significant correlation with sICAM-1 (P = 0.055), suggesting that activated endothelial cells may produce plasma nitrate. CONCLUSION: NO synthesis is elevated in SSc patients, and the activated endothelial cell is a likely site of its production. PMID- 10817564 TI - Gene therapy that inhibits nuclear translocation of nuclear factor kappaB results in tumor necrosis factor alpha-induced apoptosis of human synovial fibroblasts. AB - OBJECTIVE: Tumor necrosis factor alpha (TNFalpha) increases the survival and proliferation of human rheumatoid arthritis (RA) cell lines. These experiments were designed to determine if inhibition of nuclear factor kappaB (NF-kappaB) nuclear translocation leads to increased apoptosis of TNFalpha-treated human RA cell lines. METHODS: We constructed an inhibitor of nuclear factor kappaB(IkappaB) dominant-negative adenovirus (AdCMVIkappaB-DN) and an X-linked inhibitor of apoptosis (XIAP) antisense adenovirus (AdCMVXIAP-AS). Primary RA synovial fibroblast (RASF) cell lines were transfected in vitro, and SV40 transformed RA synovial cell lines in SCID mice were transfected in vivo. Cells were treated with TNFalpha and analyzed for apoptosis. RESULTS: There was no apoptosis of primary RASF transfected in vitro with AdCMVIkappaB-DN alone. In contrast, there was apoptosis of >85% of cells treated with AdCMVIkappaB-DN plus TNFalpha. Primary RASF in SCID mice also exhibited high levels of apoptosis after in vivo transfection with AdCMVIkappaB-DN followed by treatment with TNFalpha. There was no apoptosis after treatment with AdCMVIkappaB-DN in the absence of TNFalpha. XIAP is an inhibitor of apoptosis which was up-regulated by TNFalpha, and this up-regulation was inhibited by AdCMVIkappaB-DN plus TNFalpha. Transfection of an AdCMVXIAP-AS gene therapy resulted in increased TNFa-induced apoptosis. CONCLUSION: AdCMVIkappaB-DN gene therapy greatly enhances apoptosis due to inhibition of an NF-kappaB-mediated antiapoptosis signaling pathway, and XIAP is a TNFalpha-inducible specific inhibitor of apoptosis in RA synovial cell lines. This and other modulators of TNF receptor or the Fas apoptosis pathway may be therapeutically beneficial in facilitating apoptosis of synovial tissue in patients with RA. PMID- 10817565 TI - Differential regulation of Fas-mediated apoptosis of rheumatoid synoviocytes by tumor necrosis factor alpha and basic fibroblast growth factor is associated with the expression of apoptosis-related molecules. AB - OBJECTIVE: Fas-mediated apoptosis is associated with the pathophysiology of rheumatoid arthritis (RA). However, the molecular mechanisms of this process remain to be elucidated in rheumatoid synovium. We investigated the behavior of intracellular signaling molecules that regulate Fas-mediated apoptosis in RA synoviocytes. METHODS: Anti-Fas monoclonal antibody (mAb) was added to RA synoviocytes after treatment with tumor necrosis factor alpha (TNFalpha) or basic fibroblast growth factor (bFGF) for 5 days. The cytotoxic activity was measured using a lactate dehydrogenase-release assay. The expression of apoptosis-related molecules in RA synoviocytes was examined by immunoblot analysis. The enzymatic activities of caspases 3 and 8 under Fas ligation were examined. Transfer of the FADD (Fas-associated death domain) protein and the FLIP(L) (long form of the FLICE [FADD-like interleukin-1beta-converting enzyme]-inhibitory protein) gene into RA synoviocytes was performed using adenoviral vectors. RESULTS: Following a 5-day treatment with TNFalpha or bFGF, Fas ligation with its agonistic mAb induced apoptosis of almost all TNFalpha-treated RA synoviocytes but only showed a weak apoptotic activity in bFGF-treated synoviocytes. Although there was no correlation between the induction of Fas-mediated apoptosis and the expression of apoptosis-related molecules among these cells, a high enzymatic activity of caspases 3 and 8 was observed only in the TNFalpha-treated RA synoviocytes after Fas ligation. The bFGF-treated RA synoviocytes were relatively resistant to apoptosis induced by FADD gene transfection, as compared with the TNFalpha treated synoviocytes. In addition, the expression of FLIP(L), an inhibitory molecule of Fas-mediated apoptosis, was reduced in TNFalpha-treated RA synoviocytes, and the expression of FLIP43 was augmented in bFGF-treated RA synoviocytes. Moreover, Fas-mediated apoptosis in TNFalpha-treated RA synoviocytes was partially inhibited by FLIP(L) gene transfection. CONCLUSION: Our results suggest that Fas-mediated apoptosis of RA synoviocytes is differentially regulated by TNFalpha and bFGF. In addition, the regulatory mechanisms of apoptosis involve the formation of the death-inducing signaling complex, especially at the level of caspase 8 activation, and this process may be partly associated with FLIP expression. PMID- 10817566 TI - Regulation of synovial B cell survival in rheumatoid arthritis by vascular cell adhesion molecule 1 (CD106) expressed on fibroblast-like synoviocytes. AB - OBJECTIVE: B lymphocytes accumulate in the inflamed joints of patients with rheumatoid arthritis (RA) and are responsible for production of high amounts of (auto)-antibodies. The aim of this study was to determine the capacity of fibroblast-like synoviocytes (FLS) to contribute to the accumulation of synovial fluid (SF) B cells by extending their life span. METHODS: Highly purified SF B cells were cultured with FLS in the presence or absence of blocking antibodies directed against cell adhesion molecules, and cell viability was determined after various time intervals by trypan blue, annexin V, propidium iodide, or Hoechst staining. Phenotypic characterization of peripheral blood and SF B cells and FLS was carried out by flow cytometry. RESULTS: Synovial B cells, which consist predominantly of memory B cells and plasma cells (PC), undergo spontaneous cell death by apoptosis upon removal from their in vivo environment, despite expression of Bcl-2. Coculture with FLS rescued synovial B cells from apoptosis in a cell contact-dependent manner. Blocking studies using monoclonal antibodies demonstrated a role for the molecular interaction of SF B cells with vascular cell adhesion molecule 1 (VCAM-1; CD106) in FLS-induced survival. The ability of FLS to induce SF B cell survival was not related to the rheumatoid origin since FLS from non-RA patients had similar properties. CONCLUSION: These findings indicate a crucial role for FLS in the survival of synovial B cells at the site of inflammation in RA through the interaction with VCAM-1 expressed on FLS. Consequently, memory B cells and PC accumulation arise and persist not only as a result of maturation and recruitment of these cells, but also by active prevention from cell death by the microenvironment. PMID- 10817567 TI - Soluble vascular cell adhesion molecule 1 mediation of monocyte chemotaxis in rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is characterized by infiltration of leukocytes, including monocyte/ macrophages, into synovial tissue (ST), but factors mediating the ingress of these cells are poorly understood. Vascular cell adhesion molecule 1 (VCAM-1) plays an important role in adhesion of leukocytes to the vasculature. This study was undertaken to test the hypothesis that soluble VCAM-1 (sVCAM-1) might mediate chemotaxis of monocytes in RA. METHODS: Chemotaxis assays were performed using a modified Boyden chamber to determine the effects of sVCAM-1 on and the role of very late activation antigen 4 (VLA-4) in peripheral blood (PB) monocyte migration. Synovial fluids (SF) were immunodepleted of sVCAM 1 to identify a role for sVCAM-1 in RA. Immunohistochemistry and flow cytometry analyses were performed to show the expression of VLA-4 in ST, SF, and PB. Tyrosine phosphorylation was studied by Western blot analysis on PB monocyte lysates in the presence of signaling inhibitors. RESULTS: Soluble VCAM-1 induced monocyte migration in the nM range, in a concentration-dependent manner. Anti-VLA 4 significantly inhibited sVCAM-1-induced monocyte migration, suggesting that sVCAM-1 acts in part via a VLA-4-dependent mechanism. In RA SF, incubation with anti-VCAM-1 resulted in a reduction in the ability to induce monocyte migration (mean 28%). VLA-4 immunolocalized to RA ST, SF, or PB, monocytes, macrophages, and lymphocytes. Soluble VCAM-1 stimulated tyrosine phosphorylation in monocytes, and pertussis toxin, chelerythrine chloride, and staurosporine significantly reduced sVCAM-1-mediated monocyte chemotaxis, suggesting that signaling pathways via G proteins and protein kinase C are required for sVCAM-1-mediated monocyte migration. CONCLUSION: These results demonstrate a novel function for sVCAM-1 as a monocyte chemotactic agent in RA and suggest a new potential target for modulating monocyte ingress into inflamed RA ST. PMID- 10817568 TI - Stimulation of 92-kd gelatinase (matrix metalloproteinase 9) production by interleukin-17 in human monocyte/macrophages: a possible role in rheumatoid arthritis. AB - OBJECTIVE: To examine the cellular mechanisms by which the proinflammatory cytokine interleukin-17 (IL-17) induces the synthesis of 92-kd gelatinase (matrix metalloproteinase 9 [MMP-9]) by human monocyte/ macrophages in primary culture. METHODS: IL-17-stimulated human monocytes isolated from the peripheral blood of healthy donors were cultured in the presence of antiinflammatory cytokines, neutralizing antibodies against IL-1beta, tumor necrosis factor alpha (TNFalpha), or IL-1 receptor antagonist, and with protein kinase inhibitors of diverse specificity. MMP measurements were performed using specific enzyme-linked immunosorbent assays, while the expression of specific messenger RNA was determined by Northern blotting. Detection of phosphorylated proteins and specific transcriptional factors was performed by Western blotting and by gel retardation experiments, respectively. RESULTS: Biologically active IL-17 was detected in the synovial fluid of patients with rheumatoid arthritis. IL-17 induced MMP-9 production in human monocyte/ macrophages was dependent on endogenous prostaglandin E2 synthesis and related to autocrine stimulation by TNFalpha, but was IL-1beta independent. This activation involves both p42/44 and p38 kinases and nuclear factor kappaB. IL-17-inducible activator protein 1 and signal transducer and activator of transcription 1/3 may transactivate the MMP-9 promoter. CONCLUSION: IL-17 may contribute to an unbalanced production of proinflammatory cytokines and MMP-9 in diseased articular joint tissues by interacting with the macrophages in the rheumatoid synovium. PMID- 10817569 TI - Extracellular signal-regulated kinase 1/extracellular signal-regulated kinase 2 mitogen-activated protein kinase signaling and activation of activator protein 1 and nuclear factor kappaB transcription factors play central roles in interleukin 8 expression stimulated by monosodium urate monohydrate and calcium pyrophosphate crystals in monocytic cells. AB - OBJECTIVE: Monosodium urate monohydrate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals cause acute gout and pseudogout, respectively. Because acute gout and pseudogout appear to be dependent on interleukin-8 (IL-8)-induced neutrophil ingress, this study was undertaken to define and compare how MSU and CPPD crystals stimulate IL-8 messenger RNA (mRNA) expression in mononuclear phagocytes. METHODS: MSU and CPPD crystal-induced mitogen-activated protein kinase (MAPK) signal transduction and IL-8 transcriptional activation were studied in human monocytic cells, using the THP-1 cell line. RESULTS: MSU and CPPD crystals (0.5 mg/ml) induced activation of c-Jun N-terminal kinase, extracellular signal-regulated kinase 1 (ERK-1)/ERK-2, and p38 MAPK pathways in THP-1 cells. Activation of the ERK-1/ERK-2 pathway was essential for MSU and CPPD crystal-induced IL-8 mRNA expression, whereas the p38 pathway played a greater role in IL-8 mRNA expression in response to CPPD crystals in comparison with MSU crystals. Both crystals induced the binding of nuclear factor kappaB (NF-kappaB), including the NF-kappaB complex c-Rel/RelA, and activator protein 1 (AP-1, including N-terminal phosphorylated c-Jun) to the IL-8 promoter. Both crystals induced transcriptional activation of the IL-8 promoter, which was dependent on activation of c-Rel/RelA and AP-1. Activation of the NF-IL-6 transcription factor played a lesser role. Finally, crystal-induced IL-8 promoter activation was mediated by activation of the ERK-1/ERK-2 pathway, as demonstrated by transfection of dominant-negative raf-1. CONCLUSION: These results indicate that ERK-1/ ERK-2 signaling and transcriptional activation through AP-1 and NF-kappaB are essential for the induction of IL-8 expression in mononuclear phagocytes in response to CPPD and MSU crystals. PMID- 10817570 TI - Genetic enhancement of matrix synthesis by articular chondrocytes: comparison of different growth factor genes in the presence and absence of interleukin-1. AB - OBJECTIVE: To determine whether articular chondrocytes express growth factor genes delivered by adenoviral vectors and whether expression of these genes influences matrix synthesis in the presence and absence of interleukin-1 (IL-1). METHODS: Monolayer cultures of rabbit articular chondrocytes were infected with recombinant adenovirus carrying genes encoding the following growth factors: insulin-like growth factor 1 (IGF-1), transforming growth factor beta1 (TGFbeta1), and bone morphogenetic protein 2 (BMP-2). As a control, cells were transduced with the lac Z gene. Cultures were also treated with each growth factor supplied as a protein. Levels of gene expression were noted, and the synthesis of proteoglycan, collagen, and noncollagenous proteins was measured by radiolabeling. Collagen was typed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The effects of growth factor gene transfer on proteoglycan synthesis in the presence of IL-1 were also measured. RESULTS: The expression of all transgenes was high following adenoviral transduction. Proteoglycan synthesis was stimulated approximately 8-fold by the BMP-2 gene and 2-3-fold by the IGF-1 gene. The effects of BMP-2 and IGF-1 genes were additive upon cotransduction. Synthesis of collagen and noncollagenous proteins, in contrast, was most strongly stimulated by the IGF-1 gene. In each case, collagen typing confirmed the synthesis of type II collagen. IL-1 suppressed proteoglycan synthesis by 50-60%. IGF-1 and TGFbeta genes restored proteoglycan synthesis to control levels in the presence of IL-1. The BMP-2 gene, in contrast, elevated proteoglycan synthesis beyond control levels in the presence of IL-1. CONCLUSION: Transfer of growth factor genes to articular chondrocytes can greatly increase matrix synthesis in vitro, even in the presence of the inflammatory cytokine IL 1. This result encourages the further development of gene therapy for the repair of damaged cartilage. PMID- 10817571 TI - Hyaluronan oligosaccharides perturb cartilage matrix homeostasis and induce chondrocytic chondrolysis. AB - OBJECTIVE: To determine the role of CD44-mediated matrix assembly in maintaining cartilage homeostasis. METHODS: Articular cartilage tissue slices as well as isolated chondrocytes were treated with hyaluronan (HA) hexasaccharides. Tissues and cells were processed for histology, immunohistochemistry, colorimetric assay, reverse transcriptase-polymerase chain reaction, and zymography. RESULTS: HA oligosaccharides induced a dose-dependent state of chondrocytic chondrolysis, including near-total loss of stainable proteoglycan-rich matrix, activation of gelatinolytic activity, and exhibition of the NITEGE epitope. However, HA oligosaccharides also induced an increase of proteoglycan synthesis, including elevation of aggrecan and HA synthase 2 messenger RNA. CONCLUSION: Uncoupling chondrocytes from the matrix results in deleterious changes in matrix structure and modifications in chondrocyte metabolism. The close interaction of chondrocytes with their matrix and their ability to "sense" changes in receptor occupancy are needed for maintenance of cartilage homeostasis. PMID- 10817572 TI - Interaction of intraarticular hyaluronan and albumin in the attenuation of fluid drainage from joints. AB - OBJECTIVE: How is fluid volume regulated in joints? Fluid drainage rate is an important factor, both in normal joints and those with effusions. Hyaluronan and albumin, separately, are known to attenuate drainage, conserving synovial fluid volume in the presence of raised joint pressure. Hyaluronan and albumin normally coexist, however, in joint fluid. The objective was to determine their interactive effect on drainage. METHODS: The fluid escape rate from the joint cavity through synovium was measured at controlled intraarticular pressures using a rabbit knee model in vivo. One joint contained 4 mg/ml hyaluronan and the other contained 4 mg/ml hyaluronan plus 20 mg/ml albumin, as in normal synovial fluid. Hyaluronan-albumin interactions were assessed in vitro by viscometry and osmometry. RESULTS: Hyaluronan alone greatly attenuated fluid escape. Drainage rates plateaued at 4-5 microl/minute as pressure was raised, because the opposition to drainage increased with pressure. Addition of albumin to hyaluronan shifted the opposition-versus-pressure relation upward and further attenuated drainage by 22.5%, despite a small fall in the viscosity of the mixture. Osmometry showed a small synergistic interaction. Analysis of aspirates showed that < or =8% of albumin molecules in the draining fluid were reflected by the synovial lining (compared with 79% of hyaluronan molecules). CONCLUSION: Hyaluronan and albumin act together at normal concentrations to conserve synovial fluid in the presence of raised drainage pressures. Hyaluronan has the greater effect, acting osmotically by way of a concentration polarization boundary layer. Attenuation of this effect in arthritic effusions with low hyaluronan concentrations is one of several factors limiting fluid accumulation and, hence, the size of the effusion. PMID- 10817573 TI - Anterior crural neuralgia as a presenting manifestation of lymphocele. PMID- 10817574 TI - Clinical image: vasculitis in an old tattoo. PMID- 10817575 TI - Renal failure: a risk factor for methotrexate toxicity. PMID- 10817576 TI - Clinical images: Romanus spondylitis. PMID- 10817577 TI - Hepatitis C virus infection in patients with Sjogren's syndrome and non-Hodgkin's lymphoma: comment on the article by Voulgarelis et al. PMID- 10817578 TI - Apoptosis of CD4+ T cells in Sjogren's syndrome: comment on the article by Zeher et al. PMID- 10817579 TI - Vitamin D in corticosteroid-induced osteoporosis: comment on the article by Amin et al. PMID- 10817580 TI - Interferon-beta1a-induced juvenile chronic arthritis in a genetically predisposed young patient with multiple sclerosis: comment on the case report by Levesque et al. PMID- 10817581 TI - The effects of listening comprehension of various genres of literature on response in the linguistic area: an fMRI study. AB - Using fMRI at a static magnetic field strength of 1.5T, we investigated how comprehension and humor of sentences would correlate to activation of the language areas in listening comprehension of a native language. Sentences with a high comprehension score augmented activation in the left inferior parietal lobule and posterior part of the left superior temporal gyrus, which may be related to semantic processing. Sentences with a high humor score induced activation in Broca's area, which may be associated with syntactic processing and auditory working memory. Furthermore, sentences with a high humor factor and/or a low comprehension score activated the middle frontal gyrus, which may be attributed to auditory working memory. PMID- 10817582 TI - Multi-site partitioned delivery of human tyrosine hydroxylase gene with phenotypic recovery in Parkinsonian rats. AB - Parkinson's disease (PD) is a leading candidate for neurological gene therapy, given our increasing knowledge of the functional anatomy of the striatonigral system and the localized nature of the affected cell populations. Here we report that stereotactic introduction of a human tyrosine hydroxylase (TH-2) gene using multi-site partitioned doses resulted in behavioral recovery in 6-OHDA-lesioned rats, with transient 100% recovery observed in some animals. We also show correlation between numbers of TH-immunoreactive cells and loss of apomorphine induced rotation, with a near-linear relationship between TH expression and phenotypic recovery. Furthermore, the data suggest that only a fraction of striatal cells need to be transduced in order to exert phenotypic effects, and therefore TH partitioned gene transfer may have clinical potential in PD. PMID- 10817583 TI - Brain MRI lesions and atrophy are related to depression in multiple sclerosis. AB - It is unclear whether brain MRI lesions are associated with depression in multiple sclerosis (MS). Neurological dysfunction in depressed (n= 19) and non depressed (n = 29) MS patients was rated by expanded disability status scale (EDSS). EDSS was weakly predictive of the presence of (p = 0.03) and severity of (p = 0.01) depression. After correcting for EDSS, the presence of depression was predicted by superior frontal and superior parietal hypointense TI lesions (p<0.01); the severity of depression was predicted by superior frontal, superior parietal and temporal TI lesions, lateral and third ventricular enlargement, and frontal atrophy (p<0.01). Depression was not related to bright T2 lesions or enhancement. We conclude that atrophy and cortical-subcortical disconnection due to frontal and parietal white matter destructive lesions may contribute to depression in MS. PMID- 10817584 TI - Defects in sensory nerve numbers and growth in mutant Kit and Steel mice. AB - The roles in the nervous system of the receptor tyrosine kinase Kit and its ligand, Steel factor, are unclear. We have now found first, that sensory nerve populations are reduced in mutant Kit and Steel mice, implicating Steel-Kit interactions in neuronal development. Second, sensory axonal regeneration (which occurs independently of nerve growth factor, or NGF) is impaired, while collateral sprouting (NGF dependent) is normal. Therefore, there is a selective involvement of Kit signal transduction pathways in nerve growth; supporting this, in wild-type animals Kit was up-regulated in regenerating, but unchanged in sprouting, sensory neurons. The receptor tyrosine kinase Kit thus contrasts with the receptor tyrosine kinase trkA, which is activated by the sprouting stimulus (NGF) but not by the axonal regeneration signal. PMID- 10817585 TI - Alpha2-macroglobulin gene polymorphisms show racial diversity and are not associated with Alzheimer's disease. AB - Two genetic markers of the plasma protein alpha2-macroglobulin, a 5 bp deletion/insertion at the 5' splice site of exon 18 (A2MI) and the GTC/ATC (VaIIO00IIe) in exon 24 (A2M2), may have roles in the development of Alzheimer's disease (AD). Genotyping and linkage analysis of these markers in 426 Japanese sporadic AD patients, 85 autopsy-confirmed Caucasian AD cases, and, as controls, 382 Japanese and 65 Caucasians who were cognitively normal and 140 Japanese Parkinson's disease patients showed racial diversity in the frequencies and relationship of the two markers. Comparison of genotype and allele frequencies, stratification of the samples by the presence of the apolipoprotein E epsilon4 allele, and logistic regression analysis revealed no association of these markers with AD in either racial group. PMID- 10817586 TI - Tacrine, a reversible acetylcholinesterase inhibitor, induces myopathy. AB - Tacrine, an acetylcholinesterase (AChE) inhibitor that has been used in the treatment of Alzheimer's disease, increases available acetylcholine (Ach) levels in the synaptic cleft thereby enhancing the activity of cholinergic pathways. However, excessive stimulation of nicotinic receptors at the neuromuscular junction results in muscle deterioration. We tested whether reversible AChE inhibitors such as tacrine may induce similar effects. In the present study, tacrine administration (7.5 mg/kg twice daily) to rats produces a 20 and 30-fold increase in the number of degenerating cells in leg and diaphragm muscle, respectively, as compared to control. This myopathy is significantly decreased by co-administration of tacrine with the nitric oxide (NO) synthase inhibitor L NAME. These results show that tacrine can induce myopathy which may be mediated by increased NO production. PMID- 10817587 TI - Dynamic expression of glial cell line-derived neurotrophic factor after cerebral ischemia. AB - The aim of this study was to understand the possible involvement of glial cell line-derived neurotrophic factor (GDNF) in rat brain ischemic injury by examining the expression and the cellular location of GDNF with molecular biological and morphological techniques. Expression of GDNF mRNA and protein was first increased as early as 2h after ischemia-reperfusion in peri-infarct cerebral cortex and striatum; it then declined, and showed a second increase at 72 h. Double staining confirmed that the earlier peak of GDNF expression was of neuronal origin and the later peak of glial origin. Considering the neurotrophic characteristics of GDNF, our findings suggest that elevated endogenous GDNF expression may have important roles in protection of ischemic injured neuronal cells. PMID- 10817588 TI - Functional human heme oxygenase has a neuroprotective effect on adult rat ganglion cells after pressure-induced ischemia. AB - Adult retina subjected to transient ischemia and reperfusion leads to controlled retinal ganglion cell (RGC) death over a period. Modification of intracellular mechanisms through a specific adenoviral vector containing the hemoxygenase gene (HO-1) provides avenues for RGC survival following HO-1 gene transfer and ischemia. RGC death rate was reduced by an average of 15% at 1, 2 and 3 weeks. A significant number of RGC transfected with functional HO-1 survived ischemic insults. Pharmacological stimulation of HO-1 may constitute a novel therapeutic approach to rescuing RGC experiencing ischemic/reperfusion injury. PMID- 10817589 TI - Expression of angiotensin II AT2 receptor in the acute phase of stroke in rats. AB - A male Wistar rat model of stroke (middle cerebral artery occlusion; MCAO) was used to study the angiotensin II (Ang II) receptor subtype 2 (AT2) gene expression by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining. After permanent occlusion of the middle cerebral artery (MCA), AT2 receptor gene expression was found to increase in the infarct cortex by 2.7-fold (1 day) and 1.7-fold (3 days), respectively. Positive AT2 immunostaining was also observed in the infarct area of the cerebral cortex. Apoptotic markers were detected in the necrotic area of the stroke cerebral cortex 1 day after MCAO. This demonstrated up-regulation of AT2 receptor may be involved in the apoptosis of tissue repair after stroke. PMID- 10817590 TI - Event-related brain potentials elicited by a number discrimination task. AB - To study the number cognition process, we recorded event-related potentials (ERPs) in 14 subjects when they were indicating whether a number pair was the same or not. The two numbers given in sequence were either the same (condition 1) or different (condition 2). After 270 ms following the onset of the second stimulus in condition 2, a negative component N270 was recorded on the scalp with the most negative amplitude at the central and occipital areas. Hemispheric asymmetry was not observed in the potential. This negative component is considered to reflect the mismatching process for numbers in the brain. PMID- 10817591 TI - Rodent noradrenergic chromaffin cells contain calbindin D28K immmunoreactivity. AB - The calcium binding protein calbindin D28K is heterogeneously distributed in neurons throughout the body. We have investigated the distribution of calbindin in the chromaffin cells of the adult rodent adrenal medulla, which share the same developmental origin as peripheral sensory and autonomic neurons. Calbindin immunoreactivity was present in all noradrenergic chromaffin cells (defined by their lack of the adrenaline synthesizing enzyme, phenylethanolamine N-methyl transferase) in both the rat and mouse. It was also present in a very few adrenergic chromaffin cells in both rat and mouse. Calbindin-immunoreactivity is present in rat noradrenergic chromaffin cells from the day of birth and so is a useful marker for identifying rodent noradrenergic chromaffin cells. PMID- 10817592 TI - Developmental regulation of subunit composition of extrasynaptic NMDA receptors in neocortical neurones. AB - NMDA receptors undergo drastic changes in their subunit composition during development of the mammalian neocortex. An increase in the expression of the NR2A subunit correlates with developmental changes in the properties of synaptic NMDA receptors. In this study, we investigated whether these developmental alterations are restricted to synaptic NMDA receptors or whether similar developmental changes also occur at extrasynaptic NMDA receptors. To analyse the properties of extrasynaptic receptors, glutamate-evoked ion currents mediated by extrasynaptic NMDA receptors were isolated by irreversibly blocking synaptic NMDA receptors with MK-801. Whole-cell ion currents mediated by extrasynaptic receptors showed developmental changes in their sensitivity against the NR2B subunit-specific antagonist ifenprodil similar to that of synaptic receptors. In summary, our results strongly suggest that NR2A subunit-containing NMDA receptors increasingly contribute also to extrasynaptic NMDA receptors during in vitro differentiation. PMID- 10817593 TI - In vitro identification of dividing neuronal precursors from chick embryonic ciliary ganglion. AB - In chick parasympathetic ciliary ganglion the neuronal birthdate is well defined, between 2.5 and 5.5 days of embryonic development, and neuronal precursor cells that are able to differentiate into neurons in vitro can be isolated from E4.5 ganglia. In this report, using bromodeoxyuridine incorporation and Maplb immunostaining, we demonstrate that these cells can be isolated from E7-E8 chick embryos as well, suggesting that neuronal precursor cells are still present in the ciliary ganglion after the end of the in vivo neurogenesis. These precursor cells retain the ability to divide and generate newly differentiated neurons in vitro when cultured in a chemically defined medium. Such a capacity is highly stimulated by bFGF but not by CNTF. PMID- 10817594 TI - Allodynia and hyperalgesia evoked by sciatic mononeuropathy in NKI receptor knockout mice. AB - We have examined the participation of NK1 receptors in neuropathic pain by comparing behavioural responses after partial sciatic nerve ligation in wild-type (WT) and NK1 receptor knockout (KO) mice. Mechanical responses were tested with von Frey hairs, and cooling responses with acetone. WT and KO mice showed similar reactions before surgery. Nerve-ligated WT and KO mice showed equivalent spontaneous pain-related behaviour. Mechanical (mean threshold 20 +/- vs 9 +/- 1 mN) and cold allodynia (61 +/- vs 14 +/- 2 behaviours evoked by acetone) were significantly greater than in sham animals, but similar in WT and KO mice. We conclude that NK1 receptors are not essential for mechanical and cold allodynia evoked by partial nerve ligation. PMID- 10817595 TI - HIV-I gpI20 neurotoxicity in brain cultures is prevented by moderate ethanol pretreatment. AB - The HIV-1 coat protein gp 20, a potent neurotoxin that may underlie AIDS dementia, activates glia to cause neurotoxicity via the NMDA receptor and perhaps other routes. We find that pretreating cultures of rat organotypic cortical/hippocampal slices or cerebellar granule cells subchronically with ethanol in physiological concentrations (20-30 mM; 6 days) largely or even completely inhibits neurodegeneration due to gp120. However, NMDA-induced neurotoxicity appears unaffected by moderate ethanol pretreatment, indicating that ethanol's neuroprotection against gp120 is upstream of the NMDA receptor, possibly at a glial activation stage. The results could lead to a better understanding of relationships between ethanol, glia and neurodegeneration, particularly in AIDS. PMID- 10817596 TI - Adenosine receptor up-regulation: initiated upon preconditioning but not upheld. AB - Activation of adenosine receptors is part of the endogenous defense against cerebral hypoxia and ischemia. However, it is not known which adenosine receptor subtypes mediate hypoxic tolerance upon chemical preconditioning. Selective A3 receptor mRNA up-regulation to 135 +/- 34% (mean +/- s.d.; p<0.05) was observed 1 h after preconditioning with 3-nitropropionate while A1 receptor mRNA levels remained unchanged (94 +/- 23%; n.s.). After 24h A3 and A1 receptor mRNA expression were both at control level. Further treatment in vitro resulted in a selective A3 receptor mRNA reduction. We conclude that the early (onset within hours) but not the late (duration of days) neuroprotection upon chemical preconditioning is associated with a selective up-regulation of A3 receptor mRNA. Detection of A3 receptor mRNA is very sensitive to prolonged stress in vitro. PMID- 10817597 TI - Association of the N-acetyltransferase I gene (NATI) with mild and severe substance abuse. AB - We observed a significant increase in the number subjects carrying the NAT1* 10 allele of the N-acetyl transferasel (NAT1) gene in controls with a MAST-R score of > or = 4 and in subjects with drug and/or alcohol dependence (p=0.003), compared with controls with a MAST-R <4. These results suggest that alterations in the acetylation of one or more CNS compounds may be related to both mild and severe substance abuse. PMID- 10817598 TI - Carrier-mediated transport and enzymatic hydrolysis of the endogenous cannabinoid 2-arachidonylglycerol. AB - The human astrocytoma cell line CCF-STTGI accumulates [3H]2-AG through an Na(+)- and energy-independent process, with a Km of 0.7 +/- 0.1 microM. Non-radioactive 2-AG, anandamide or the anandamide transport inhibitor 4-hydroxyphenyl arachidonamide inhibit [3H]2-AG uptake with half-maximal inhibitory concentrations (IC50) of 5.5 +/- 1.0 microM, 4.2 +/- 0.3 microM and 1.8 = 0.1 microM, respectively. A variety of lipid transport substrates and inhibitors interfere with neither [3H]2-AG nor [3H]anandamide uptake. These results suggest that 2-AG and anandamide are internalized in astrocytoma cells through a common carrier-mediated mechanism. After incubation with [3H]2-AG, radioactivity is recovered in phospholipids, monoacylglycerols (unmetabolized [3H]2-AG), free fatty acids ([3H]arachidonate) and, to a minor extent, diacylglycerols and triacylglycerols. Arachidonic acid (100 microM) and triacsin C (10 microM), an acyl-CoA synthetase inhibitor, prevent incorporation of [3H]arachidonic acid in phospholipids and significantly reduce [3H]2-AG transport. Thus, the driving force for 2-AG internalization may derive from the hydrolysis of 2-AG to arachidonate and the subsequent incorporation of this fatty acid into phospholipids. PMID- 10817599 TI - Attention to touch modulates activity in both primary and secondary somatosensory areas. AB - We used fMRI to establish whether attention to touch enhances somatosensory cortical activity. Subjects received somatosensory and visual stimulation and were instructed to attend selectively to one modality during alternating stimulus detection periods interspersed with rest periods during which no stimulus was delivered. The maximum signal change for each task versus rest was measured in anatomically defined regions of interest for each subject. Attended touch produced greater signal change than unattended touch in primary (S1) and secondary (S2) somatosensory cortex. In contrast to the conclusions of some previous studies, we found that the enhancement of activation with attention was at least as great in S1 as in S2. The attentional effect was unilateral in S1 and bilateral in S2 and the somatosensory insula. PMID- 10817600 TI - Low frequency damped oscillations of cat visual cortical neurones. AB - A synchronizing volley of excitation was delivered to cells from the primary visual cortex of the anaesthetized cat by an electrical pulse stimulation of the optic radiations. In intracellular records, this resulted in an initial, fast depolarization of the membrane potential followed by a longer lasting (266 ms) hyperpolarization, and then to small amplitude damped oscillations of the membrane potential. In different cells the number of peaks ranged from 3 to 6, and the amplitude of the first oscillation averaged 3.5 mV. If the pulse stimulation was repeated during visual drive with an optimal bar stimulus, the membrane potential fluctuations were non-linearly enhanced with spike bursts recorded during the depolarized states of the membrane potential. The amplitude of the first oscillation in these cases averaged 6.9 mV. The mean frequency of damped oscillations was 9.9 Hz. The results suggest that these oscillations are mediated by the waxing and waning of the excitatory drive to the recorded neurone. PMID- 10817601 TI - Survival and regeneration of motoneurons in adult rats by reimplantation of ventral root following spinal root avulsion. AB - The present study examines whether reimplantation of the ventral root could prevent motoneuron death after root avulsion. In the control animals about 65% or 39% of motoneurons survived at 3 or 6 weeks post-injury respectively. More than 60% of them expressed nitric oxide synthase (NOS). In contrast, in animals with ventral root reimplantation, nearly 90% or 80% of motoneurons survived at 3 or 6 weeks post-injury respectively. Expression of NOS due to root avulsion was significantly inhibited in these experimental animals. More interestingly, about 80% of the surviving motoneurons were found to regenerate their axons into the reimplanted ventral root, and all of these regenerating motoneurons were NOS negative. Results of the present study show that reimplantation of avulsed ventral root can greatly enhance motoneuron survival and the surviving motoneurons can regrow their axons into the original ventral root. PMID- 10817602 TI - Synaptic efficacy is commonly regulated within a nervous system and predicts individual differences in learning. AB - The hypothesis that an individual's capacity for learning might be predicted or influenced by basal levels of synaptic efficacy has eluded empirical tests, owing in part to the inability to compare between animals single identified synaptic responses in the mammalian brain. To overcome this limitation, we have focused our analysis on the invertebrate Hermissenda, whose nervous system is composed of identifiable cells and synaptic interactions. Hermissenda were exposed to paired presentations of light and rotation such that the light came to elicit a learned defensive motor response. An animal's rate of learning was strongly correlated with the amplitude of the synaptic potential evoked in that animal's visual (light sensitive) receptors in response to stimulation of presynaptic vestibular (rotation sensitive) hair cells. In naive animals, strong correlations between the amplitude of both inhibitory and excitatory synaptic potentials were observed between synapses distributed throughout an animal's nervous system, and this conservation of synaptic efficacy was largely attributable to a common influence on transmitter release. These observations suggest that basal synaptic efficacy may be uniformly regulated throughout a nervous system, and provide direct evidence that the basal efficacy of synaptic transmission predicts, and possibly contributes to, individual differences between animals in their capacity to learn. PMID- 10817603 TI - Distribution of SNAP-25 in transient neuronal circuitries of the developing human forebrain. AB - The distribution of SNAP-25 is demonstrated within prominent transient structures in the developing human forebrain. During early fetal development SNAP-25 is mainly expressed in axons of the intermediate zone and the internal capsule. The fibers appear directed towards the mantle zone of the ganglionic eminence and the perireticular nucleus located within the internal capsule. Cells of these two areas are shown to interact with SNAP-25 immunoreactive structures with the aid of double-labellings. The SNAP-25 immunoreactive fibers may represent corticofugal axons which contact the perireticular nucleus and ganglionic eminence which are regarded as intermediate targets providing a scaffold for growing axons. Anti-SNAP-25, thus, is an appropriate marker of intermediate targets which are involved in brain injuries of preterm infants. PMID- 10817604 TI - Inhibition of neuronal nitric oxide synthase protects against MPTP toxicity. AB - Previous work showed that several relatively specific inhibitors of neuronal nitric oxide synthase (nNOS) produce protection against MPTP induced dopaminergic toxicity. We examined whether a highly specific novel inhibitor of nNOS, ARRI 7338, could also protect against MPTP toxicity. ARR17338 produced dose-dependent significant protection against MPTP induced depletion of dopamine and protected against MPTP induced depletions of tyrosine hydroxylase immunostained neurons in the substantia nigra. These results provide further evidence that inhibitors of nNOS may be useful for the treatment of Parkinson's disease. PMID- 10817605 TI - Sedative effects of the dopamine D1 receptor agonist A 68930 on rat open-field behavior. AB - The present results demonstrate sedative effects of the DA D1 receptor agonist A 68930 (0.9-15 micromol kg(-1), s.c.) on rat spontaneous locomotor activity in an open field. The effects were particularly strong, and dose-dependent, for the ambulatory activity in the open-field arena (forward locomotion) and for rearing activity, whereas the suppression of locomotor activity (i.e. total horizontal activity in the open field) was less conspicuous. The distribution of activity within the open field (activity in center vs periphery) was not consistently affected by the A 68930 treatment. In support for DA D1 receptor mediated effects of A 68930, the effects on locomotor activity, forward locomotion, and on rearing behavior, were partially antagonized by the DA D1 receptor antagonist SCH 23390 (15 nmol kg(-1) s.c.). SCH 23390 by itself produced a modest, but statistically significant, suppression of these different items of open-field behavior. The atypical antipsychotic agent clozapine has previously, in this laboratory, been shown to stimulate DA D1 receptors in vivo. There are a number of clinical and laboratory observations, consistent with the notion of a beneficial role for such effects in schizophrenia. Thus, the sedation, apparently not related to extrapyramidal motor functions, produced by DA D1 receptor agonist A 68930 could reflect an important aspect of the mechanism of action for atypical antipsychotic drugs. PMID- 10817606 TI - Nitric oxide synthase activity and immunoreactivity in the crayfish Procambarus clarkii. AB - The presence, characteristics and localization of nitric oxide synthase (NOS) in the eyestalk ganglia of the crayfish (Procambarus clarkii) were investigated. Biochemical assay indicated that NOS activity was significantly inhibited by N(G) nitro-L-arginine (a NOS inhibitor), fluphenazine-N-2-chloroethane (a calmodulin inhibitor) as well as by omitting beta-NADPH or Ca2+ from the incubations. Immunohistochemical studies localized NOS immunoreactivity (IR) to the first chiasma and the sinus gland. Finally, a NOS-immunoreactive protein (138 kDa) was detected by Western blot analysis. These data demonstrate for the first time the existence in crustacean eyestalk ganglia of NOS with biochemical characteristics similar to mammalian constitutive NOS. The localization of NOS-IR suggests that NO may be involved in regulating visual processes and neuroendocrine function in crustaceans. PMID- 10817607 TI - Eye position effects in macaque area V4. AB - Recent studies revealed an influence of eye position on neuronal discharges in many dorsal stream areas of the macaque visual cortical system. This eye position information is thought to serve an implicit non-retinocentric representation of visual spatial information. The question arises of whether the two visual cortical pathways encode information in a common coordinate system, i.e. whether eye position effects exist also in ventral stream areas. We recorded 112 neurons from area V4 of two awake monkeys. Of these, 55 (49%) showed eye position effects. Like in dorsal stream areas, this modulatory influence was balanced out at the population level. Our data support the view of eye position effects as a general phenomenon in the macaque visual cortical system. PMID- 10817608 TI - Effects of passive tactile co-activation on median ulnar nerve representation in human SI. AB - In animals simple passive co-activation causes a fusion and expansion of the involved cortical representations. We used passive tactile finger co-activation for 40 min to investigate cortical representational changes in the human somatosensory cortex. Magnetic source imaging revealed that the euclidean distance between median and ulnar nerve somatosensory evoked fields (SEF) was significantly reduced after application of 600 synchronous airpuff stimuli to the fingertips of four fingers. In the control experiment without co-activation no significant change in distance was observed. Perception threshold and spatial two point discrimination were not affected by the synchronous stimulation. This is in contrast to blind three-finger Braille readers who frequently mislocalize stimuli applied to the reading fingers. This points to a lack of behavioural relevance or the short duration of co-activation. PMID- 10817609 TI - Differential effects of pain and spatial attention on digit representation in the human primary somatosensory cortex. AB - Reorganization of primary somatosensory cortex subsequent to either reduced or enhanced peripheral input is well established. Recently, plastic changes following arm amputation in humans were shown to correlate with phantom limb pain. This raised the question whether spatial attention and pain may cause cortical reorganization in the absence of deafferentation. Using non-invasive neuroelectric imaging to study the digit representation in the human primary somatosensory cortex, we report a delayed shift of the representation of digits 2 3 due to pain on the digits 4-5, which outlasted the pain by several minutes. In contrast, reorganization during spatial attention was less pronounced, was seen almost immediately and only during the condition. These data indicate that spatial attention and pain without peripheral deafferentation cause cortical reorganization by different mechanisms. The differential time course of reorganizational effects observed at the cortex may be due to modulation of the lemniscal pathways by nociceptive input from the spinal cord dorsal horn. PMID- 10817610 TI - Double-pulse stimulation dissociates intrathalamic and cortical high-frequency (>400Hz) SEP components in man. AB - Human somatosensory evoked potentials (SEP) contain high-frequency (600 Hz) wavelet bursts possibly reflecting repetitive population spikes in thalamocortical axons and/or postsynaptic responses. To dissociate thalamic and cortical burst components the recovery of intrathalamic SEP (derived from electrodes implanted for movement disorder therapy in seven patients) was compared with scalp SEP in six age-matched Parkinsonian patients and six healthy younger subjects. Upon electric median nerve double-pulse stimulation conditioned scalp bursts were found attenuated in both groups, more for 10ms than 20ms interstimulus intervals; moreover, intraburst frequencies decreased from 690Hz to 590Hz. By contrast, intrathalamic burst amplitudes and frequencies (around 1 kHz) remained largely stable. These dissociations indicate functionally distinct generator mechanisms for scalp and intrathalamic high-frequency SEP bursts. PMID- 10817611 TI - Tapping movements according to regular and irregular visual timing signals investigated with fMRI. AB - Whole-head functional MR images were acquired while 10 subjects were asked to tap with their right index finger in synchrony with a visual stimulus appearing regularly with a frequency of 1.5 Hz, or irregularly with a mean frequency of 1.5 Hz. Performance data show that during regular tapping most taps were close to stimulus onset. However, when the subjects paced their tapping according to the irregular stimuli, most taps appeared about 300 ms after the onset of the pacing stimuli. Comparing the brain activations resulting from regular tapping with those from irregular tapping, we found increased activation in left precuneus only. Comparing irregular versus regular tapping shows increased activity in right cerebellar nuclei and vermis, left ventrolateral thalamus, left sensorimotor cortex, left and right pre-SMA and left SMA proper. These results show that during irregular pacing the motor areas are more strongly activated than during regular pacing. In addition, further neural systems are involved in the motor control during irregular pacing: cerebellar vermis and a cerebello thalamo-cortical system. The latter is supposedly involved in error correction in the context of visually guided movements. PMID- 10817612 TI - Absence of TrkB and insulin receptor beta in the Triton insoluble low-density fraction (raft). AB - Cholesterol- and glycolipid-enriched microdomains within the plasma membrane of animal cells, including neurons, have been purified and used as a low-density membrane domain after extraction with Triton X-100 (raft), or after subcellular fractionation without detergent (LDM). In this study, we compared the protein compositions in the raft and the LDM. Membrane receptors, acylated- and glycosylphosphatidylinositol (GPI)- anchored proteins were enriched in the LDM. Further treatment of the LDM with Triton X-100 excluded the membrane receptors, TrkBs and insulin receptor beta. In the presence of calcium ions, the endogenous tyrosine kinase activities in the LDM and the raft were enhanced, suggesting an important role of calcium ions in the signal transduction via the LDM and the raft. PMID- 10817613 TI - Stimuli outside the classical receptive field modulate the synchronization of action potentials between cells in visual cortex of cats. AB - It is proposed that various attributes of an image are bound neuronally when responsive units fire in synchrony. Our investigations describe the influences of the contextual stimuli upon the occurrence of synchronization, in anaesthetized cats. Once a significant synchronization was recorded in the cross-correlogram (XCRG) between evoked action potentials of two groups of neurons in response to a drifting sine-wave grating, additional gratings were positioned outside the compound receptive field. The synchronization strength was then measured in relation to the difference between the orientations of the central and peripheral gratings. In the majority of cases results indicate that the synchronization is facilitated with larger orientation disparities. Thus, our data support the notion that contrasting features of images facilitate synchrony of activity between neurons. PMID- 10817614 TI - Increment of synapsin I immunoreactivity in the hippocampus of the rat kindling model of epilepsy. AB - Synapsin is a vesicle-associated protein that is thought to be involved in synaptogenesis and neurotransmitter release. In this study, we investigated the repeated amygdala kindling-induced changes in levels of synapsin I, a marker of enhanced synaptic activity, using immunohistochemistry. Eight hours after the last seizure, synapsin I immunoreactivity (IR) was increased bilaterally in the polymorphic zone of the dentate gyrus, the hilus and the stratum lucidum in area CA3 of the hippocampus. However, 4 weeks after the last kindled seizures, synapsin I IR was not significantly changed in any of the brain regions studied. Therefore, the increased levels of synapsin I protein observed in these brain regions early after the last seizure may reflect the plastic changes that occur in the specific neural networks of the hippocampus directly after kindling induced seizure activity. PMID- 10817615 TI - Effect of neurokinin-I receptor antagonists on the function of 5-HT and noradrenaline neurons. AB - Substance P antagonists have been proposed to be a new class of antidepressants. The present study aimed to determine the effect of the selective non-peptide rat neurokinin-1 (NK1) receptor antagonists WIN 51,708 and CP-96,345 on the firing activity of rat dorsal raphe serotonin (5-HT) and locus coeruleus noradrenaline (NA) neurons. While WIN51,708 (2mg/kg, i.v.) and CP-96,345 (0.15 mg/kg, i.v.) did not modify the firing activity of 5-HT and NA neurons, both antagonists attenuated the suppressant effect of the alpha2-adrenoceptor agonist clonidine on the firing activity of both types of neurons. In contrast, the responsiveness of 5-HT neurons to the i.v. administration of the 5-HT autoreceptor agonist LSD and the 5-HT1A receptor agonist 8-OH-DPAT remained unchanged. These findings suggest that NK1 receptor antagonists affect markedly the NA system via an attenuation of the function of alpha2-adrenoceptors on the cell body of NA neurons and, consequently, may also modulate 5-HT neurotransmission. PMID- 10817616 TI - The time-course of intermodal binding between seeing and hearing affective information. AB - Intermodal binding between affective information that is seen as well as heard triggers a mandatory process of audiovisual integration. In order to track the time course of this audiovisual binding, event related brain potentials were recorded while subjects saw facial expression and concurrently heard auditory fragment. The results suggest that the combination of the two inputs is early in time (110 ms post-stimulus) and translates as a specific enhancement in amplitude of the auditory NI component. These findings are compatible with previous functional neuroimaging results of audiovisual speech showing strong audiovisual interactions in auditory cortex in the form of magnetic response amplifications, as well as with electrophysiological studies demonstrating early audiovisual interactions (before 200 ms post-stimulus). Moreover, our results show that the informational content present in the two modalities plays a crucial role in triggering the intermodal binding process. PMID- 10817617 TI - Functional neuroanatomy of smooth pursuit and predictive saccades. AB - We used PET to study differences in cerebral blood flow (CBF) in smooth pursuit, predictive saccades and fixation. Eye movements were monitored in the scanner. Compared with fixation, pursuit and predictive saccades activated a network of highly similar areas, including frontal eye fields, supplementary eye fields, V5 and medial cuneus. Our findings are consistent with non-human primate studies that suggest that pursuit and saccades are controlled by similar and adjacent neural areas. Pursuit was associated with greater activation of caudate than saccades, suggesting a role for basal ganglia in pursuit that is consistent with studies of neurological populations. Saccades were associated with greater activation of cerebellum and frontal eye fields. A frontal-cerebellar loop may be important in coordinating the preparation and timing of saccades in predictive tracking. PMID- 10817618 TI - Neuroprotective potency of kynurenic acid against excitotoxicity. AB - The aim of this study was to determine in vivo which extracellular levels of kynurenic acid (KYNA) are required to control excessive NMDA receptor activation in the rat cortex. As excitotoxicity is coupled to marked ion movements, local depolarisations induced by perfusion of NMDA or quinolinic acid (QUIN) through microdialysis probes were recorded at the site of excitotoxin application. Perfusion of KYNA through the dialysis fibre inhibited the excitotoxin responses with an IC50 of 32-66 microM (extracellular concentration corrected for microdialysis delivery), but > 10-fold lower levels of endogenous KYNA were reported to be neuroprotective. Accordingly, these results strengthen the notion that KYNA accumulation may protect the brain parenchyma by acting on different molecular target(s), besides the NMDA receptor glycine site. PMID- 10817619 TI - The precuneus in motor imagery: a magnetoencephalographic study. AB - Magnetoencephalography was applied to subjects who imagined themselves hurdling in self-centered space. In three of six subjects all 300 trials in the motor imagery condition revealed the precuneus dipole. When we divided the 300 trials into four overlapping blocks (one block = 150 trials), all six subjects showed precuneus activity. The latency of the precuneus dipole was about 220 ms. We suggest that the precuneus activity during motor imagery involves retrieval of spatial information and/or setting up spatial attributes. Only in one subject but twice, the current dipole located in the supplementary motor area was observed 60 ms after activation of the precuneus, which suggests that the signal from the precuneus for motor imagery is transferred to the supplementary motor area. PMID- 10817620 TI - HIV-derived protein gp120 suppresses P3 potential in rats: potential implications in HIV-associated dementia. AB - Between 20 and 30% of AIDS patients have neurological symptoms characterized by motor impairment, memory loss and progressive dementia. Previous studies have implicated the HIV derived gp120, which produces behavioral deficits and electrophysiological alterations in rats. The goal of the present study was to describe the effect of this protein on the P3 event-related potential (ERP), evoked by a passive discrimination task in rats. We used II rats divided into two groups: HIV gp120 (n = 6) and control (n = 5). We recorded the P3 wave before any treatment (baseline), during the i.c.v. administration of either HIVgp 120 (700 ng/5 days) or saline (pH 7.2), and 24 h, 7, 14 and 21 days after the last injection. There were no changes between groups in the amplitude or latencies of the observed components (N1, P2, N2 and P3) evoked by target stimuli, during baseline or during the injection period. However, the HIV gp120 group showed a significant amplitude reduction in P3 wave 24 h after the last injection, while the N1, P2 and N2 waves remained unchanged. However, from the 7th day through the 21st day, P2 and N2 components also disappeared and only the N1 component could be observed in the HIV gp 20-treated group. These changes in the N2, P2 and P3 potentials, suggesting an alteration in cognitive processes, further support the neurotoxic activity of HIV gp120 and its role in AIDS dementia. PMID- 10817621 TI - BN 80933 inhibits F2-isoprostane elevation in focal cerebral ischaemia and hypoxic neuronal cultures. AB - Formation of the lipid peroxidation product 8-epi-prostaglandin2alpha (8-epi PGF2alpha) a bioactive marker of oxidative stress, was quantified in in vitro and in vivo models of neuronal death. In culture media of primary rat cortical neurones exposed to hypoxia followed by reoxygenation, a 3.7-fold increase of 8 epi-PGF2alpha concentration was observed in comparison to control cells. In rats submitted to 2h middle cerebral artery occlusion followed by a 22h reperfusion period, a 27-fold increase of 8-epi-PGF2alpha was observed in the ischaemic hemisphere compared with the corresponding hemisphere of sham-operated rats. Treatment with the neuroprotective agent BN 80933 significantly reduced both 8 epi-PGF2alpha elevations in vitro and in vivo. These data suggest that 8-epi PGF2alpha elevations might reflect the damaging free radical overproduction and subsequent lipid peroxidation during neuronal injury induced by hypoxia and ischaemia. Inhibition of 8-epi-PGF2alpha elevations participates to the neuroprotective effects of BN 80933. PMID- 10817622 TI - Dramatic brain aminergic deficit in a genetic mouse model of phenylketonuria. AB - Clinical data suggest that brain catecholamines and serotonin are deficient in phenylketonuria (PKU), an inherited metabolic disorder that causes severe mental retardation and neurological disturbances. To test this hypothesis, brain tissue levels of dopamine (DA), norepinephrine (NE), 5-hydroxytryptamine (5-HT) and their metabolites were evaluated in the genetic mouse model of PKU (Pah(enu2)). Results indicated a significant reduction of 5-HT levels and metabolism in prefrontal cortex (pFC), cingulate cortex (Cg), nucleus accumbens (NAc), caudate putamen (CP), hippocampus (HIP) and amygdala (AMY). NE content and metabolism were reduced in pFC, Cg, AMY and HIP. Finally, significantly reduced DA content and metabolism was observed in pFC, NAc, CP and AMY. In pFC, NAc and CP there was also a marked reduction of DA release. PMID- 10817623 TI - IGF-I expression is decreased in LIF-deficient mice after peripheral nerve injury. AB - We investigated the regulation of insulin-like growth factor 1 (IGF-1) expression after sciatic nerve crush using leukemia inhibitory factor (LIF)-deficient mice. One day post-crush, IGF-1 mRNA levels were lower in the LIF-deficient mouse nerve than in the wild type nerve. IGF-1 protein, analyzed by immunohistochemistry, was also decreased 1 day post-crush in LIF-deficient nerves relative to wild type nerves. By 3 days post-crush, IGF-1 immunoreactivity was induced in Schwann cells to equivalent levels in both types of nerve. After crush, IGF-1 expression was also found in mast cells, and these were initially decreased in the LIF-deficient mice. Thus, LIF appears to regulate IGF-1 expression in the peripheral nerve basally and early in the regeneration response in vivo. PMID- 10817624 TI - Mapping the differences in the brain concentration of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in an animal model of depression. AB - Antidepressant drugs as well as electroconvulsive stimuli can significantly influence brain concentrations of neurotrophic factors. However, it is not known whether the baseline brain concentrations of neurotrophic factors are altered in human subjects suffering from affective disorders or whether there are sex differences in concentrations of neurotrophins in human brain. In order to elucidate some of these questions, we measured by ELISA brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in an animal model of depression, the Flinders Sensitive Line (FSL) rats and their controls, the Flinders Resistant Line (FRL). Altered BDNF and NGF concentrations were found in frontal cortex, occipital cortex, and hypothalamus of depressed FSL compared to FRL control rats. Furthermore, different levels of these neurotrophins were also found in the male and female brain. Cumulatively these observations suggest that BDNF and NGF may play a role in depression and, hypothetically, different brain regional concentrations of BDNF and NGF in male and female animals may be relevant to gender differences in vulnerability to depression. PMID- 10817625 TI - Combined effect of exposure to lead and chlordane on the testicular tissues of swiss mice. AB - Chlordane, is one of many environmental pollutants that reach human or animal body through food and water consumption, its presence may frequently be associated with other metals such as lead. In this work, the toxicity of chlordane and lead to reproductive tissues in Swiss mice was studied. Oral daily doses of 75 and 275 mg/kg b.w. were administered to male mice, for 35 days, animals were sacrificed at the 2nd, 3rd, 4th, and 5th week. The results show that chloradane had a damaging effect on testicular tissues, the effect was obvious through reduction in diameter of the seminiferous tubules, number of spermatogonia, and primary and secondary spermatocytes and spermatids. The effect was remarkably increased by the presence of lead, which needs more investigation in order to determine whether it's a synergistic or additive effect. PMID- 10817626 TI - Absence of structural or functional alterations in male and female reproductive organs of F1 and F2 generations derived from female mice exposed to 3'-azido-3' deoxythymidine during pregnancy. AB - To investigate the effects of in utero exposure to 3'-azido-3'-deoxythymidine (AZT) on male and female reproductive system development, pregnant CD-1 mice were given daily intragastric doses of 25.0 mg AZT during days 12 through 18 of gestation. The offspring were examined at birth, as well as at pubertal, young adult and adult stages of development, for reproductive organ endpoints including anogenital distance, onset of testicular descent, latency to vaginal opening, and proportion of time for each of the stages of estrous cycle. These reproductive endpoints remained mostly unchanged in AZT-treated offspring as compared to the controls. Males and females exposed in utero to AZT (F1 generation) were fertile when mated to untreated females and males, respectively, and their liveborn F2 offspring showed no adverse effects for any of the reproductive parameters tested. Thus, no evidence of developmental reproductive toxicity was noted either in the F1 mice exposed to AZT during the critical period of male and female reproductive system development, or in the F2 mice born of matings between the AZT-exposed F1 mice and unexposed animals. PMID- 10817627 TI - Polymorphisms of the gene for microsomal epoxide hydrolase and susceptibility to alcoholic liver disease and hepatocellular carcinoma in a Caucasian population. AB - The gene encoding the xenobiotic-metabolising microsomal enzyme, epoxide hydrolase (mEPHX), shows two common mutations, i.e. at exons 3 and 4. It is unknown how these genetic polymorphisms relate to risk of developing alcoholic liver disease (ALD) and/or hepatocellular carcinoma (HCC) in a Caucasian population. DNA samples extracted from the blood of 61 ALD patients and 203 healthy controls, and from archival liver tissue of 46 cases of HCC, were subjected to polymerase chain reaction amplification followed by digestion with EcoR V or Rsa I to demonstrate polymorphisms of exon 3 or 4, respectively. The distributions of the genotypes of exon 3 in the ALD and HCC patients, and exon 4 in the HCC patients did not differ significantly from those of the control group. However, compared with the control group, the ALD group contained a significantly greater number of individuals homozygous or heterozygous for the exon 4 mutation. This suggested association between possession of the exon 4 mutant mEPHX allele and increased risk of developing ALD may relate to known interactions between mEPHX and alcohol-metabolising enzyme systems, or to linkage disequilibrium between the mutation and other genetic risk factors for ALD. PMID- 10817628 TI - Effects of soaking temperature and soaking time during preparation of water extract of tea on anticlastogenicity against environmental tobacco smoke in the sister-chromatid exchange assay. AB - Water extract of tea (WET) was prepared by soaking green tea at different temperatures for various periods of time and was used to test whether the soaking temperature and soaking time during the preparation of WET influence the content of polyphenols and the anticlastogenicity of WET against environmental tobacco smoke (ETS). Five major polyphenols in WET were measured. Extractable-respirable particulate (ERP) was obtained from ETS-contaminated indoor air (ERP-ETS). The sister-chromatid exchange assay (SCE) was utilized to evaluate the clastogenic effects of ERP-ETS and the anticlastogenic effects of WET. The results indicate that ERP-ETS is clastogenic and WET has significant anticlastogenic effects on ERP-ETS. The content of polyphenols and the anticlastogenic potential of WET depended on the soaking temperature and soaking time during WET preparation. At the soaking temperature of 80 degrees C, an increased soaking time was correlated with a higher percentage of polyphenols and a concomitantly enhanced anticlastogenic efficacy. By contrast, at the soaking temperature of 100 degrees C, a longer soaking time was associated with a higher percentage of polyphenols concomitant with a lower anticlastogenic efficacy. The data suggest that, besides polyphenols, and additional material(s), which may be partially inactivated at 100 degrees C, is contributing to the anticlastogenicity of WET. PMID- 10817629 TI - Capture ELISA and flow cytometry methods for toxicologic assessment following immunization and cyclophosphamide challenges in beagles. AB - The purpose of this subacute 22-day study was to evaluate methods for canine circulating immunoglobulins (IgM, IgG, and IgE) and select B- and T-lymphocyte populations (CD4-helpers, CD8-suppressors, pan-T and pan-B) for immunotoxicity testing using an organ system (concordance) approach. The challenge substance for immunoglobulin testing was repeated immunization with six-way distemper vaccination (DHLAPP), while the challenge substance for leukocyte subpopulations was treatment with cyclophosphamide. Immunoglobulin measurements were made by capture enzyme-linked immunosorbent assay (ELISA), and leukocyte immunophenotyping by fluorescein isothiocyanate/phycoerythrin conjugation (flow cytometry). A battery of parameters that would be used in a typical regulatory study were taken to aid interpretation of the data generated by these methods. Body weights, food consumption, clinical observations, complete clinical chemistry and urinalysis measurements were taken. Gross pathology and micropathology of sternal bone marrow, spleen, mesenteric and retropharyngeal lymph nodes, thymus, liver and kidney were completed. The ELISA method demonstrated acceptable intra-assay reproducibility for IgM, IgG and IgE, with values in good agreement as reported for radial immunodiffusion. The immunologic challenge demonstrated a biological trend of an increase in IgM that preceded an increase in IgG with no discernible trend in IgE response, and no abnormalities in lymphocyte subpopulations. Principle flow cytometry findings related to cyclophosphamide were that the relative percent of B cells decreased dramatically and progressively after compound administration; being statistically decreased in males on day 22 compared with day -5. The relative percent CD4 and CD8 contribution increased, but the CD4/CD8 ratio remained relatively unchanged as total white blood cells decreased progressively. The increase in relative percent CD4 (males only) was statistically significant according to a two-sample t-test on days 17, 20 and 22 when compared with the pre-treatment day -5. There was a relative percent increase in CD5-panT, but absolute numbers were dramatically decreased. We conclude that an organ system approach to assessment of the immune system which incorporates humoral antibody, enumeration of lymphocyte populations and pathologic evaluation of the lymphoreticular organs assists in the interpretation of an adverse toxicological response. The ELISA method for measurement of Igs detected the expected levels of IgG, IgM and IgE due to repeated vaccinations and to cyclophosphamide treatment. The flow cytometry method was acceptable for measuring select canine lymphocyte populations and detecting the expected decrease in B cells due to cyclophosphamide treatment. Both methods may be added to a testing battery for assessing immunotoxicityl in canine regulatory studies. PMID- 10817630 TI - Prolonged phenobarbital pretreatment abolishes the early oxidative stress component induced in the liver by acute lindane intoxication. AB - Lindane administration to rats (60 mg/kg b.w.) led to an enhancement in the oxidative stress status of the liver at 4 h after treatment, characterized by increases in hepatic thiobarbituric acid reactants (TBARS) formation and chemiluminescence, reduced glutathione (GSH) depletion, and diminution in the biliary content and release of GSH. These changes were observed in the absence of changes in either microsomal functions (cytochrome P450 content, NADPH-dependent superoxide radical production, and NADPH-cytochrome P450 reductase or NADPH oxidase activities) or in oxidative stress-related enzymatic activities (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glucose-6-phosphate dehydrogenase, and glutathione-S-transferases), over control values. Phenobarbital (PB) administration (0.1% in drinking water; 15 days) elicited an enhancement in liver microsomal functions, lipid peroxidation, and GSH content, without changes in oxidative stress-related enzymatic activities, except for the elevation in those of glutathione reductase and glutathione-S transferase, compared to control rats. Lindane given to PB-pretreated rats did not alter liver microsomal functions, lipid peroxidation, glutathione status, or oxidative stress-related enzymatic activities, as compared to PB-pretreated animals. In addition, lindane induced periportal necrosis with hemorrhagic foci in untreated rats, but not in PB-pretreated animals. It is concluded that the early oxidative stress response of the liver to lindane and hepatic injury are suppressed by PB pretreatment via induction of microsomal enzymes in all zones of the hepatic acinus. reserved. PMID- 10817631 TI - Photocytotoxic and DNA damaging effect of temoporfin (mTHPC) and merocyanine 540 (MC540) on nasopharyngeal carcinoma cell. AB - Photodynamic therapy (PDT) is a new approach to cancer treatment for a variety of malignant tumors. In this study, two clinical photosensitizers, Temoporfin (meta tetra-hydroxyl-phenyl-chlorin; mTHPC) and merocyanine 540 (MC540), were selected to explore for their photocytotoxic and genotoxic effects on nasopharyngeal carcinoma cells (NPC/HK1 and CNE2). Results of tetrazolium reduction assay showed that 80% cell killing were achieved for both cell lines at 0.4 microg/ml mTHPC for 24 h incubation and then with 40 kJ/m2 light irradiation, whereas 40 microg/ml MC540 with 50 kJ/m2 light dosage was required to attain the same level of phototoxicity for NPC/HK1. On the contrary, NPC/CNE2 was quite resistant to MC540. Hence, mTHPC-mediated PDT exerted a more potent effect than MC540-mediated PDT, even though the molar extinction coefficient of the main absorption peak for MC540 is much higher than that of mTHPC. Dark cytotoxicity remained negligible for both sensitizers. Comet assay was used to evaluate the DNA strand break and potential genotoxic effect induced by mTHPC and MC540 on the NPC cells. No DNA strand break was detected in the absence of light, and under sublethal treatment (LD25) for either sensitizer-loaded cells. Confocal laser scanning microscopy showed that mTHPC and MC540 localized in the cytoplasm but not in the nucleus of the tumor cells, which provided evidence for undetectable DNA damage under dark and low photodynamic dose. PMID- 10817632 TI - Rat extracorporeal circulation model for evaluation of systemic immunotoxicity. AB - We have applied a rat extracorporeal circulation (EC) model as an evaluation system for the immunotoxicity of medical devices in contact with the blood stream. Combining popular hemodialysis (HD) membranes [a non-biocompatible membrane, Cupurophane (CUP), and more biocompatible membranes, Cu-ammonium rayon (CAR) and polyacrylonitrile (PAN)] with rat EC, we evaluated the elicitation of acute and delayed immunological responses, as well as the effect of repeated EC. Acute effect markers such as the production of tumor necrosis factor (TNF)-alpha and complement activity during EC, and delayed effect markers such as beta2 microglobulin (beta2-M), IgG, and complement 3 levels, were monitored. Acute markers after EC passage showed responses similar to those previously reported in patients with long-term hemodialysis such as TNF-alpha production and increased complement activity. Although beta2-M and IgG levels increased to 3- to 5-fold of the initial concentration within 4 weeks after rat EC, the trend of IgG increase was inversely correlated with membrane biocompatibility (CUP > CAR = PAN), but this did not occur for elevations in beta2-M (PAN > CAR > CUP). These data suggest that this EC model can reproduce similar immunological responses as seen in HD patients, and can be employed to evaluate medical devices and materials for their delayed, systemic, and repeated exposure effects with respect to immunotoxicity. PMID- 10817633 TI - CYP 450 enzyme induction by chronic oral musk xylene in adult and developing rats. AB - Developmental and adult toxicity of musk xylene was studied in Long Evans (LE) rats fed with chow containing musk xylene (MX) in food pellets in concentrations of 1 mg, 10 mg, 33 mg, 100 mg and 1000 mg MX per 1 kg chow corresponding to a daily intake of 0.07-0.08 mg MX/kg up to 70-80 mg MX/kg body weight. Adult male and female rats were MX exposed for a minimum of 10 weeks before mating. Exposure continued throughout pregnancy, birth and lactation. The effects of MX on CYP1A1/1A2 were studied in liver microsomes by EROD (7-ethoxyresorufin-rosomes deethylase) for CYP1A1 and by MROD (methoxyresorufin-o-demethylase) for CYP1A2 activity and by Western blotting. MX induced these enzymes dose dependently in adult and developing rats at PN (postnatal day) 1 and 14. The lowest effective maternal dose was 2-3 mg MX/kg/day. Western blot data of CYP2B and CYP3A indicated the induction of both P450 enzyme proteins in developing rats at PN 14 at the higher dose of 70-80 mg MX/kg/day. In contrast, upon high MX exposure CYP2B but not CYP3A was found to be induced in adult first generation male and female rats, indicating differential sensitivity to MX in development. PMID- 10817634 TI - The potential role of rodents in the enzootic cycle of Rift Valley fever virus in Senegal. AB - Wild rodents (214) of fourteen species were trapped at seven sites in Senegal. Arvicanthis niloticus and Mastomys erythroleucus were among the most frequently collected species (77.2% of total capture). All rodents were examined for the presence of anti-Rift Valley fever virus (RVFV) antibody; the prevalence over all sampled species was 3.8%, varying widely with respect to species and location. Four of 14 species of rodents were found to have anti-RVFV antibodies: Rattus rattus (one positive of two tested) Mastomys huberti (13.5%), A. niloticus (4.3%), and M. erthroleucus (2.4%). The highest prevalence of anti-RVFV antibody was recorded within the enzootic area of the Senegal River delta, at Richard Toll (9.6%). A. niloticus and M. erythroleucus and a strain of laboratory-bred mice were experimentally inoculated with two strains of RVFV and examined for viremia, illness, seroconversion and mortality. A. niloticus and M. erythroleucus demonstrated a limited resistance to infection, thus potentially allowing for the replication of virus in these animals and making these species possible candidates as hosts in the maintenance cycle of RVFV in nature. PMID- 10817635 TI - Acute Trypanosoma cruzi infection is associated with anemia, thrombocytopenia, leukopenia, and bone marrow hypoplasia: reversal by nifurtimox treatment. AB - In the present work we show that acute infection of C3H mice with the CL strain of Trypanosoma cruzi is characterized by an exponential growth of parasites and high mortality accompanied by anemia, thrombocytopenia, leukopenia, and bone marrow hypoplasia. Administration of nifurtimox, a trypanocydal drug currently in clinical use at different days postinfection, modulates parasitemia and prevents mortality. More importantly, none of blood and bone marrow alterations were observed in nifurtimox-treated animals when treatment was initiated early in infection, one or seven days postinoculation. The bone marrow alterations were characterized by a decrease in the total number cells as well in the number of megakaryoblasts and erythroblasts. Transfer experiments of bone marrow cells from infected mice to noninfected lethally irradiated recipients revealed a poor marrow-repopulating activity. The colony forming units-spleen assay confirmed the depression of committed clonal progenitors cells and revealed a decreased number of granulocyte/macrophage, megacariocyte and erythrocyte colonies. In summary, this is the first report showing that acute T. cruzi infection results in profound alterations of the hematopoietic system and that these alterations can be prevented by nifurtimox treatment. PMID- 10817636 TI - Genetic heterogeneity of HIV-1 in Greece. AB - The aim of this study was to detect and determine the genetic variation of HIV-1 in Greece and to analyze the phylogenetic relationships and transmission dynamics of identified variants. Eighty-six blood samples from HIV-1 seroconverted patients of different risk groups were collected from the AIDS clinic, AHEPA Hospital, Thessaloniki, Greece. Retroviral DNA was extracted from uncultured peripheral blood mononuclear cells. HIV-1 DNA sequences encoding a 500-bp fragment of the gp120 C2-C3 region were amplified from each study subject, and they were genetically subtyped by heteroduplex mobility assay and DNA sequencing. Genetic distances and phylogenetic relationships of DNA sequences were estimated using PHYLIP software. Our results revealed that 82 out of 86 (95.3%) subjects carried subtype B sequences, while four (4.7%) carried subtype A sequences. Subtype A in Greek individuals not having traveled abroad was documented. An average of intrasubtype B genetic divergence of 15% was noted. Our findings demonstrate the presence of at least two genetic subtypes of HIV-1 in northern Greece--subtype B and subtype A. The predominant subtype is subtype B, which was transmitted into Greece by multiple sources. Our observations lend support to the argument that the distribution of HIV-1 subtypes is determined by founder effects or other processes rather than any tropism for particular cell types or mode of transmission. PMID- 10817637 TI - Immunocytochemistry of the AfaE adhesin and AfaD invasin produced by pathogenic Escherichia coli strains during interaction of the bacteria with HeLa cells by high-resolution scanning electron microscopy. AB - We used a recent scanning electron microscope equipped with field emission gun and highly sensitive detectors to develop a fast and simple protocol for double immunogold staining using 10- and 15-nm gold particles. We used this approach to analyse the afimbrial adhesive sheath produced by pathogenic Escherichia coli interacting with the surface of epithelial cells. We demonstrated that AfaE adhesin and AfaD invasin were exposed at the bacterial surface during the interaction. This method could be easily and widely extended to the study of the early invasion process of many bacterial and viral pathogens, by immunocytochemical probing. PMID- 10817638 TI - Type III secretion system in Chlamydia species: identified members and candidates. AB - Chlamydia trachomatis and Chlamydia pneumoniae genomes contain genes coding for type III secretion apparatuses. Like other pathogens, Chlamydia probably uses this system to secrete proteins in the host cell. With the aim of identifying such proteins, we analyzed the organization of Chlamydia type III secretion genes. PMID- 10817640 TI - The role of interleukin 15 in mounting an immune response against microbial infections. AB - Interleukin (IL)-15 is a recently described cytokine that resembles IL-2 in its biological activities, stimulating natural killer cells, T cells and B cells to proliferate, secrete cytokines, and exhibit increased cytotoxicities or produce antibody. IL-15 also exerts unique functions such as stimulation of phagocytes, maintenance of mast cells and migration of activated/memory T cells. IL-15 is involved in protection against infections with a variety of microbes through not only activating innate immunity but also mounting adaptive immunity. PMID- 10817639 TI - Natural killer cell surveillance of intracellular antigen processing pathways mediated by recognition of HLA-E and Qa-1b by CD94/NKG2 receptors. AB - HLA-E binds specifically to MHC class Ia leader peptides in a TAP (transporter associated with antigen processing)-dependent manner. It interacts with CD94/NKG2A receptors on natural killer cells and this inhibits natural killer cell lysis of the cell displaying HLA-E. The crystal structure of HLA-E demonstrates that the specificity of leader peptide binding is a structurally defined intrinsic property of HLA-E. PMID- 10817641 TI - Contribution of the hydrophobic effect to microbial infection. AB - The hydrophobic effect has been known for decades. Numerous researchers have invoked the hydrophobic effect to explain how pathogens adhere to tissues. In some cases, inhibition of adhesion can be brought about by low concentrations of aromatic compounds, such as p-nitrophenol or tryptophan. Because the hydrophobic effect has been considered to be nonspecific, the molecular biology of adhesive hydrophobins has not been studied in as much detail as lectin adhesins. The literature provides compelling evidence that a large number of bacterial and fungal pathogens depend on hydrophobic interactions for successful colonization of a host. Several laboratories are now developing effective antiadhesins, based on inhibition of hydrophobic interactions between the host and the pathogen. PMID- 10817642 TI - Protein phosphatase 2A: a definite player in viral and parasitic regulation. AB - Cells use phosphorylation/dephosphorylation mechanisms to regulate the activity of several proteins required to transmit information from the cell surface to the nucleus. Recent studies have significantly increased our knowledge regarding the structure/function of one major regulator of cell phosphorylation: protein phosphatase 2A (PP2A). This review will discuss the role of PP2A in virology and parasitology. PMID- 10817643 TI - Sepsis: the critical role of iron. AB - Sepsis is a global problem which is exacerbated by increasing bacterial resistance to antibiotics. However, mechanisms of natural resistance can be extremely effective, and need to be exploited, but the availability of iron is critical for controlling bacterial growth. The diagnosis of sepsis and possible strategies for limiting iron availability are discussed. PMID- 10817644 TI - Q fever epidemiology and pathogenesis. AB - The lungs are a port of entry and primary infectious focus of Coxiella burnetii, the obligate intracellular contagium of the worldwide zoonosis Q fever. The infectious process and immune response are characterised by studies in cell culture and animal systems. Following endocytosis, replication exclusively occurs in the phagolysosome. Several potential virulence factors are described. PMID- 10817645 TI - Generation of MHC-peptide tetramers: a new opportunity for dissecting T-cell immune responses. AB - In the past few years, the technical breakthrough in generating MHC-peptide tetramers has revolutionized the analysis of T-cell responses. The major advantage of this technique over currently available methods is the ability of these tetramers to label T lymphocytes according to their antigenic specificity. The present review describes some technical aspects of tetramers generation and discusses some of the numerous possibilities opened up by this new technology. PMID- 10817646 TI - Historical perspectives on the etiology of tuberculosis. AB - Robert Koch's 1882 demonstration that the tubercle bacillus was the true cause of tuberculosis established a new understanding of causation in medicine. This scientific breakthrough set in motion an etiological revolution with vast implications for the control of infectious disease, and its ramifications are still being felt today. PMID- 10817647 TI - 'Infectious web'. AB - The current web summary highlights internet sites describing the growing problem of Staphylococcus aureus antibiotic resistance, the epidemiology and prevention of malaria, biochemical and genetic aspects of lipopolysaccharide action, and the history and treatment of hepatitis C infections. PMID- 10817648 TI - Noonan syndrome: a cryptic condition in early gestation. AB - Noonan syndrome is one of the most common of genetic syndromes and manifests at birth, yet it is usually diagnosed during childhood. Although prenatal diagnosis of Noonan syndrome is usually not possible, in a few cases the ultrasonographic findings suggested the diagnosis in utero. Reported sonographic clues include septated cystic hygroma, hydrothorax, polyhydramnios, and cardiac defects, such as pulmonic stenosis and hypertrophic cardiomyopathy. During a 6-year period, 46,224 live-born infants were delivered at the Chaim Sheba Medical Center. Seven newborn infants and four fetuses were found to have Noonan syndrome. One fetus showed transient nuchal translucency of 4 mm and bilateral neck cysts at the 13th gestational week. Both findings resolved spontaneously by the 18th gestational week, but during the third trimester this fetus developed hydrothorax, skin edema, and polyhydramnios. In the three other fetuses, first- and second trimester ultrasonographic findings were normal, and the diagnosis of Noonan syndrome was suggested only during the third trimester. All three fetuses had polyhydramnios and skin edema. A cardiac malformation, hydrothorax, and a large head were present in one fetus. Sonographic facial findings were investigated. In all four fetuses posteriorly angulated, apparently low-set ears and depressed nasal bridge were identified. Wide nasal base was seen in two fetuses. In two fetuses, persistent opening of the fetal mouth was interpreted as fetal hypotonia. One fetus developed progressive postnatal hypertrophic cardiomyopathy and in one case, pulmonic stenosis became apparent at age 6 months. This small series suggests that Noonan syndrome has an evolving phenotype during in utero and postnatal life. Amelioration of early nuchal region findings and late onset of the more "typical" ultrasonographic changes may limit early prenatal detectability. PMID- 10817649 TI - Imperforate anus in Feingold syndrome. AB - A father and daughter had the characteristic findings of Feingold syndrome including microcephaly, short palpebral fissures, brachydactyly with clinodactyly of fifth fingers, and bilateral syndactyly of second to third and fourth to fifth toes. The infant presented with long-gap esophageal atresia without fistula (type A). Her father, who had short stature and learning disabilities, had congenital imperforate anus with a recto-vesical fistula. This is the first report of distal intestinal atresia in Feingold syndrome. PMID- 10817650 TI - Mutations at the ataxia-telangiectasia locus and clinical phenotypes of A-T patients. AB - Mutations at the ataxia-telangiectasia (A-T) locus on chromosome band 11q22 cause a distinctive autosomal recessive syndrome in homozygotes and predispose heterozygotes to cancer, ischemic heart disease, and early mortality. PCR amplification from genomic DNA and automated sequencing of the entire coding region (66 exons) and splice junctions detected 77 mutations (85%) in 90 A-T chromosomes. Heteroduplex analysis detected another 42 mutations at the A-T locus. Out of a total of 71 unique mutations, 50 were found only in a single family, and 51 had not been reported previously. Most (58/71, 82%) mutations were frameshift and nonsense mutations that are predicted to cause truncation of the A T protein; the less common mutation types were missense (9/71, 13%), splicing (3/71, 4%) and one in-frame deletion, 2546 3 (1/71, 1%). The mean survival and height distribution of 134 A-T patients correlated significantly with the specific mutations present in the patients. Patients homozygous for a single truncating mutation, typically near the N-terminal end of the gene, or heterozygous for the in-frame deletion 2546 3, were shorter and had significantly shorter survival than those heterozygous for a splice site or missense mutation, or heterozygous for two truncating mutations. Alterations of the length or amino acid composition of the A-T gene product affect the A-T clinical phenotype in different ways. Mutation analysis at the A-T locus may help estimate the prognosis of A-T patients. PMID- 10817651 TI - Anticipation in inflammatory bowel disease: a phenomenon caused by an accumulation of confounders. AB - Inflammatory bowel disease (IBD) has a definite genetic component as documented by epidemiological and linkage evidence. It shows an earlier onset of disease in children of affected patients than in their parents. This has lead to speculations about genetic anticipation in this disorder. 2,007 IBD patients with sporadic disease and 472 multiplex familial cases (including 103 affected parents and 99 children of affected patients) were evaluated with a multi-item questionnaire as part of a study of inflammatory bowel disease genetics. The Mann Whitney U-test and the general linear model were used for analysis. Clinical characteristics such as presence of fistulae, stenoses, extraintestinal manifestations, and other parameters, which are related to the severity of the disease, were found to be similar between familial and sporadic cases of IBD (corrected P > or = 0.31 for all tests). The mean-age-of onset in children of affected patients was 19.4 years earlier than in their parents. However, the age of the parental cohort was significantly higher (27 years) and the diagnostic interval also longer (1.7 years). If these confounders are corrected in a general linear model, no significant difference is evident for the age-of-onset between the groups (P > or = 0.52). There is no evidence for genetic anticipation in inflammatory bowel disease. The absence of genetic anticipation is consistent with the clinical similarity of familial and sporadic inflammatory bowel disease. This finding justifies the primary genetic analysis of familial disease under the assumption that their genetic background will be representative for all presentations of IBD. PMID- 10817652 TI - Haplotype-phenotype correlation in Fukuyama congenital muscular dystrophy. AB - In typical Fukuyama congenital muscular dystrophy (FCMD), peak motor function is usually only unassisted sitting or sliding on the buttocks, though a few patients are able to walk at some point. However, a few patients have a severe phenotype and never acquire head control. In addition, it is clinically difficult to differentiate this severe FCMD from Walker-Warburg syndrome (WWS) or from muscle eye-brain disease (MEBD). In order to establish a genotype-phenotype correlation, we performed haplotype analysis using microsatellite markers closest to the FCMD gene (FCMD) in 56 Japanese FCMD families, including 35 families whose children were diagnosed as FCMD with the typical phenotype, 12 families with a mild phenotype, and 9 families with a severe phenotype. Of the 12 propositi with the mild phenotype, 8 could walk and the other 4 could stand with support; 10 cases were homozygous for the ancestral founder (A-F) haplotype whereas the other 2 were heterozygous for the haplotype. In the 9 severe cases, who had never acquired head control or the ability to sit without support, 3 had progressive hydrocephalus, 2 required a shunt operation, and 7 had ophthalmological abnormalities. Haplotype analysis showed that 8 of the 9 cases of the severe phenotype are heterozygous for the A-F haplotype, and the other one homozygous for the haplotype. We confirmed that at least one chromosome in each of the 56 FCMD patients has the A-F haplotype. The rate of heterozygosity for the A-F haplotypes was significantly higher in severe cases than in typical or mild cases (P < 0.005). Severe FCMD patients appeared to be compound heterozygotes for the founder mutation and another mutation. Thus, the present study yielded molecular genetic evidence of a broad clinical spectrum in FCMD. PMID- 10817653 TI - Congenital hypertrichosis, cardiomegaly, and osteochondrodysplasia (Cantu syndrome): a new case with unusual radiological findings. AB - We report on a new case of a syndrome first described by Cantu et al. [1982: Hum Genet 60:36-41] comprising congenital hypertrichosis, "coarse" facial appearance, and mild osteochondrodysplasia. Our case has some unusual radiological findings, namely proximal and distal megaepiphyses of long bones and advanced bone age. PMID- 10817654 TI - Girl with accelerated growth, hearing loss, inner ear anomalies, delayed myelination of the brain, and del(22)(q13.1q13.2). AB - We report on an 18-month-old Japanese girl with 46,XX,del(22)(q13.1q13.2). To our knowledge, this is the first report of a case of interstitial deletion of a 22q13.1-q13.2 segment. Clinical features included hearing loss accompanied by inner ear anomalies, hypotonia and minor anomalies, such as a long philtrum, full eyelids, epicanthus, left transverse palmar crease and psychomotor developmental delay. Despite the chromosomal deletion, her physical growth was accelerated: her height was between the 75th and 90th percentiles for her age. Her brain MRI showed signs of delayed myelination. The three-dimensional MRI of the inner ear showed abnormalities of the cochlea and vestibule in both ears. Clinical features of the patient are similar to those of a patient with a del(22)(q13.1q13.33) karyotype previously reported by Romain et al. PMID- 10817655 TI - Clavicular hypoplasia, zygomatic arch hypoplasia, and micrognathia: a newly defined syndrome. AB - We report on a 6-year-old boy with a previously undefined syndrome of clavicular hypoplasia, frontonasal malformation, zygomatic arch hypoplasia, micrognathia, and normal intelligence. His condition differs from similar syndromes on the basis of unique facial findings such as microcornea, stellate irises, and a midline maxillary cleft. We present his case, a review of the literature, and propose the acronym CHZAM, for clavicular hypoplasia, zygomatic arch, and micrognathia, to represent this syndrome. PMID- 10817656 TI - Association between historically high frequencies of neural tube defects and the human T homologue of mouse T (Brachyury). AB - The human T developmental gene has been implicated in the etiology of neural tube defects (NTDs) on the basis both of mouse studies of its homologue, T (Brachyury), and of allelic association in a Caucasian population. We have investigated the frequency of the T allelic variant TIVS7-2 in 218 Irish NTD case parent triads. This population showed the same trend as previously reported, with an excess of the TIVS7-2 allele among cases. Log-linear modeling of case and maternal genotypic effects within families indicated that TIVS7-2 was elevated in cases (relative risk, RR = 1.36) but not in mothers (RR = 0.91). The TIVS7-2 allele is markedly associated with cases born before 1980 (RR = 2.09; CI = 1.23 3.55; corrected p = 0.030), but not with more recent cases (RR = 0.92). Cases carrying a TIVS7-2 allele did not show any increased tendency to be homozygous for the thermolabile variant of the folate-dependent enzyme 5,10-methylene tetrahydrofolate reductase, which is an established genetic risk factor for NTDs. Since the incidence of NTDs has declined markedly in Ireland over the last few decades, we suggest that the T-associated risk is potentiated by nutritional or environmental risk factor(s), the impact of which have been diminishing over time. PMID- 10817657 TI - Major gene segregation of actinic prurigo among North American Indians in Saskatchewan. AB - Actinic prurigo is an idiopathic, familial photodermatosis seen especially in American Indians. Segregation analysis was performed on 12 Saskatchewan pedigrees with American Indian ancestry, comprising a total of 1,148 individuals, ascertained via probands diagnosed with actinic prurigo. Although a high degree of familial aggregation has been noted in the past and dominant inheritance has been suggested, no formal segregation analysis has been attempted. Actinic prurigo has a variable age of onset and, therefore, age at the time of censoring must be taken into account in the analysis. However, as these ages of 57% of the unaffected individuals were missing, an algorithm was devised to impute the missing ages from known birth years in the family based on the age differences among relatives and spouses. Using these imputed ages, simple dominant inheritance with incomplete penetrance and a single age of onset distribution was found. The method for imputing the ages at examination was evaluated, as was the correction for ascertainment, by using alternative methods and comparing the results. Regardless of the method used, a dominant mode of inheritance without any multifactorial component remained the best hypothesis. PMID- 10817658 TI - Genetic fine mapping of the gene for nonsyndromic congenital retinal nonattachment. AB - Autosomal recessive nonsyndromic congenital retinal nonattachment (NCRNA) comprises congenital insensitivity to light, massive retrolental mass, shallow anterior chamber, microphthalmia, and nystagmus in otherwise normal individuals. Polymerase chain reaction-based linkage analyses of polymorphic microsatellite markers in the 10q21 region on DNA samples from 106 individuals provide evidence that the NCRNA locus is within an interval of approximately 0.6-1.5 cM, flanked by the markers D10S522 and D10S1418. Haplotype analysis demonstrated a unique founder haplotype shared by 100% of the NCRNA chromosomes. These results indicate a founder effect and the strong possibility of a single mutation as the cause of the disease in the affected population. Based on these findings, it is now possible to provide relatively accurate carrier detection and prenatal diagnostic testing for families with NCRNA based on close flanking markers and the capacity to identify NCRNA chromosomes by their haplotypes. PMID- 10817659 TI - Growth charts for young children with neurofibromatosis 1 (NF1) PMID- 10817660 TI - Effect of cadmium or magnesium on calcium-dependent central function that reduces blood pressure. AB - The effect of intracerebroventricular (i.c.v.) administration of cadmium or magnesium on central calcium-dependent blood pressure regulation was investigated. The systolic blood pressure of spontaneously hypertensive rats (SHR; male, 13 weeks of age) decreased following i.c.v. administration of cadmium chloride (20 nmol/rat), and increased following i.c.v. administration of magnesium chloride (20, 600, and 1,200 nmol/rat). The hypotensive effect of cadmium was suppressed by i.c.v. administration of W-7 (a calmodulin antagonist, 30 microg/rat). Taking into consideration these results with our previous reports, it is suggested that cadmium binds to the calcium-binding sites of calmodulin and activates calcium/calmodulin-dependent enzymes in a disorderly manner, whereas magnesium does not. Therefore, cadmium increases dopamine synthesis in the brain via a calmodulin-dependent system, and the resultant increase in dopamine levels inhibits sympathetic nerve activity and reduces blood pressure in SHR. PMID- 10817661 TI - Evidence that the reactions of nickel in the presence of vitamin C do not produce toxic oxygen intermediates such as hydroxyl but ascorbate and carbon radicals. AB - A variety of reactions containing different amounts of nickel ions together with vitamin C were prepared and used in spin trapping experiments designed to show nickel is not a Fenton active metal. Carbon based radicals were observed at low ratios of ascorbate to nickel, and ascorbate radical was observed at high ratios of ascorbate to nickel. No buffer effects were observed. There was no evidence for oxygen intermediates as products of the reaction with 5,5-dimethyl-1 pyrroline N-oxide (DMPO) or alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN), seeming to indicate nickel is not a Fenton metal. To test this hypothesis further trapping studies were run with the singlet oxygen trap 2,2,6,6-tetramethyl-4 piperidone hydrochloride, in which dioxygen was found not to affect the formation of carbon radicals. These results help to explain the damage to DNA observed in the presence of vitamin C and nickel in vitro. PMID- 10817662 TI - Role of high-energy phosphates and their metabolites in protection of carbofuran induced biochemical changes in diaphragm muscle by memantine. AB - A combined antidotal treatment with memantine HCI (MEM, 18 mg/kg, s.c.) and atropine sulfate (ATS, 16 mg/kg, s.c.) provided complete protection against acute carbofuran toxicity (1.5 mg/kg, s.c.) in rats by multiple mechanisms. Carbofuran, in addition to inhibiting serine-containing esterases, also perturbed the activities of mitochondrial/cytoplasmic biomarker enzymes (creatine kinase, CK; and lactic dehydrogenase, LDH) in diaphragm muscle. The observed changes in the activity of biomarker enzymes were reflected in serum as a result of their leakage from the diaphragm due to a depletion of high-energy phosphates, such as adenosine triphosphate (ATP, 27%) and phosphocreatine (PCr, 33%) and their metabolites (ADP, 36%; AMP, 35%; and Cr, 38%). Combined treatment with MEM and ATS provided protection and reversal of the induced changes in biomarkers by preventing depletion of high-energy phosphates and thus maintaining normal cell membrane characteristics, including permeability and integrity. These results, along with those reported previously, indicate that MEM antagonizes carbofuran toxicity by multiple mechanisms. PMID- 10817663 TI - Effect of human plasma on the reactivation of sarin-inhibited human erythrocyte acetylcholinesterase. AB - The reactivation of organophosphate-inhibited acetylcholinesterase (AChE) by oximes inevitably results in the formation of highly reactive phosphoryloximes (POX), which are able to re-inhibit the enzyme. In this study, the dependence of POX formation on AChE concentration was investigated with sarin-inhibited human erythrocyte AChE (EryAChE). A marked dependence was found with obidoxime but not with the experimental oxime HI 6, suggesting great differences in the decomposition rates of the respective POXs. At a physiological erythrocyte content the reactivation of EryAChE was markedly affected by POX with obidoxime and pralidoxime (2-PAM) but not with the newer oximes HI 6 and HLo 7. Addition of extensively dialysed, sarin-treated human plasma reduced the reactivation by obidoxime and 2-PAM even more. Obidoxime and 2-PAM were superior to HI 6 and HLo 7 in reactivating butyrylcholinesterase (BChE). This effect was pronounced in diluted plasma, but was obscured in concentrated plasma, probably because of re inhibition by the generated POX. Addition of native erythrocytes to sarin-treated plasma resulted in marked inhibition of EryAChE in the presence of obidoxime, suggesting a higher affinity of the POX for EryAChE. The results indicate that obidoxime and 2-PAM may reactivate sarin-inhibited AChE insufficiently due to re inhibition by the POX formed. In addition, the re-inhibition of Ery-AChE may be aggravated by the POX that is produced during BChE reactivation. These reactions must be regarded as therapeutically detrimental and disqualify those oximes which are capable of forming stable POX by reactivation of BChE. PMID- 10817664 TI - The phosphoryl oxime-destroying activity of human plasma. AB - The potential of obidoxime and other pyridinium-4-aldoximes to reactivate dimethyl- and diethylphosphorylated cholinesterases is markedly restricted by the inevitable formation of rather stable phosphoryl oximes (POXs) with high anticholinesterase activity. This effect is hardly seen with very dilute enzyme preparations, but becomes significant at physiological enzyme concentrations. Human plasma with the butyrylcholinesterase irreversibly blocked by soman was able to stimulate obidoxime-induced reactivation of concentrated erythrocyte acetylcholinesterase (Ery-AChE) to the same extent as was observed with a dilute preparation, suggesting phosphoryl oxime-destroying capacity. The inactivating factor, which was tentatively termed POX-hydrolase, had (1) a molecular weight of >100 kDa; (2) required Ca2+ , which could not be substituted by Zn2+ or Mg2+; and (3) lost its catalytic activity reversibly in the presence of ethylenediamine tetraacetic acid (EDTA). The enzyme activity varied widely (20-fold) among different subjects and did not follow the activity pattern of human serum paraoxonase (PON1). Rabbit plasma with its particularly high paraoxonase content showed only weak POX-hydrolase activity. These data suggest POX-hydrolase to be a different entity. POX-hydrolase was most active with the putative phosphoryl obidoxime from paraoxon-ethyl, less with the product from paraoxon-methyl and least with that from diisopropylfluorophosphate. The analogue TMB-4 reacted similarly to obidoxime. The putative phosphonyl oximes arising by the reaction of obidoxime with nerve agents were apparently not cleaved. The variation in POX hydrolase activity may additionally contribute to the variable response to oxime therapy in patients with organophosphate insecticide poisoning. PMID- 10817665 TI - Induction and inhibition of testicular germ cell apoptosis by fluoroacetate in rats. AB - Fluoroacetate (FA), an inhibitor of aconitase, is known to lower the intracellular level of adenosine triphosphate (ATP), which recently has been suggested to be a possible determinant of the form of cell death, apoptosis or necrosis. To investigate which form of germ cell death occurs in FA-induced testicular toxicity, adult Sprague Dawley rats were given a single oral dose of FA (0.5 or 1.0 mg/kg) and euthanized at 3, 6, 12, 24, 48, and 72 h thereafter. Germ cell degeneration was histologically first found in early round spermatids at stage I and in spermatogonia at stages II-IV of seminiferous tubules 6 and 12 h, respectively, after dosing. Degenerating spermatogonia exhibited characteristic features of apoptosis as demonstrated by both electron microscopy and in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), whereas spermatids did not. At the 24 and 48 h time points, degenerating spermatids were continually present and subsequently formed multinucleated giant cells, while the number of degenerating spermatogonia and TUNEL-labeled spermatogonia was drastically and/or significantly decreased compared to those from the control group, indicating that spontaneous male germ cell apoptosis is inhibited. Coincident with these morphological changes, DNA laddering on gel electrophoresis was apparent only 12 h after dosing. The results demonstrate that FA induces either apoptosis or necrosis of male germ cells in the early stage after dosing and subsequently inhibits spontaneous apoptosis. PMID- 10817667 TI - Changes in serum alpha2u-globulin levels in male rats given diethylstilbestrol and applicability to a screening test for endocrine-disrupting chemicals. AB - alpha2u-Globulin (AUG) is a major rat urinary protein, which has a molecular weight of 16 kDa (kidney type) or 19 kDa (native type). The biosynthesis of this protein is under multi-hormonal regulation. In this study, we investigated changes in serum AUG level and their association with changes in the reproductive organs of male rats after the administration of the estrogenic chemical, diethylstilbestrol (DES) at doses ranging from 0.01 mg/kg per day to 100 mg/kg per day by gavage for 14 days. Our aim was to establish basic data for the development of a new screening method for endocrine disrupting chemicals based on serum AUG levels. DES treatment decreased the weight of testes in a dose dependent manner; and was accompanied by atrophic histopathological changes in testes. Testis weights were significantly decreased by the group given 1 mg/kg per day DES; however, histopathological abnormalities were found in the group given 0.1 mg/kg per day DES. In four of five animals in the group given 1 mg/kg per day there was no significant decrease in testis weight and only a slight or moderate degeneration of the pachytene spermatocytes. Despite these findings, serum AUG levels in this group decreased markedly, while the serum AUG level markedly decreased even in the animals with no histopathological change in the 1 mg/kg per day or 0.1 mg/kg per day groups with no histopathological change also showed decreased serum AUG level. These results suggest that the serum AUG level may be a sensitive parameter for detecting the activity of estrogenic chemicals in intact male rats. Although a uterotropic assay has been proposed for immature female or ovariectomized female rats and is currently undergoing validation studies internationally, there is no screening method for estrogenic chemicals in intact male animals. More data on AUG changes by treatment with other estrogenic chemicals are needed in order to determine the sensitivity and specificity of this response to estrogens. Nonetheless, an AUG-based screening test for estrogenic chemicals may be useful owing to its applicability to conventional toxicity studies and an apparently higher sensitivity of this parameter compared to organ weight change or histology of testis in intact male rats and applicability to conventional toxicity studies. PMID- 10817666 TI - Cytotoxicity of capsaicin in monkey kidney cells: lack of antagonistic effects of capsazepine and Ruthenium red. AB - Capsaicin is a natural product of Capsicum peppers, excitatory effects of which have been shown to be mediated by the recently cloned vanilloid receptor 1 (VR1). Since previous studies have shown that capsaicin inhibits protein synthesis, experiments were performed to investigate whether this effect is mediated by VR1 receptor on cultured monkey kidney cells (Vero cells). The capsaicin uptake was assessed in cellular homogenate and in medium by high-performance liquid chromatography (HPLC) separation and quantification on C18 reverse-phase column and fluorescence detection. Toxic effects were assessed by incorporation of [3H]L leucine into cellular proteins in the presence of capsazepine, the VR1 vanilloid receptor antagonist and Ruthenium red or tyrosine or calcium. Capsazepine (1 to 256 microM) did not modify the uptake rate of capsaicin for incubation times up to 24 h and did not antagonize capsaicin-induced protein synthesis inhibition. It rather inhibited protein synthesis per se from 100 to 256 microM. Ruthenium red which blocks mitochondrial calcium uptake, inhibited protein synthesis and did not antagonise or increase synergistically the effects of capsaicin. Interestingly in a medium deprived of calcium and supplemented by calcium chloride (10-50 microM) the protein synthesis inhibition induced by capsaicin is antagonised somehow. There was no prevention of capsaicin diffusion into the cells. Tyrosine, which seems to be the best preventive agent of capsaicin inhibitory effects, prevents its metabolism but not its diffusion. Capsaicin might enter cells by diffusion and interfere with protein synthesis machinery by competition with tyrosine which in turn prevents the metabolism of capsaicin. The results of the present study suggest that cell responses to capsaicin may be transduced through at least two molecular pathways, one involving VR1, since the receptor antagonist capsazepine fails to prevent the inhibitory effect of capsaicin in Vero cells of renal origin. PMID- 10817668 TI - Lack of oxidative DNA damage or initiation of carcinogenesis in the kidneys of male F344 rats given subchronic exposure to p-dichlorobenzene (pDCB) at a carcinogenic dose. AB - p-Dichlorobenzene (pDCB) is a male rat kidney carcinogen believed to act through alpha2u-globulin nephropathy. Recent data on metabolism, however, suggest a potential for generating oxidative stress. To examine possible mechanisms of kidney carcinogenesis, pDCB was studied for ability to produce 8 oxodeoxyguanosine (8-oxodG) in kidney nuclear DNA and for initiating activity in a two-stage renal carcinogenesis model. F344 male rats were given pDCB by intragastric instillation, 5 days/week for 13 weeks at 300 mg/kg per day, which is a carcinogenic dose with chronic administration. To assess initiation after exposure, trisodium nitrilotriacetic acid (NTA), a kidney tumor promoter was given in the drinking water at 1,000 ppm for 39 weeks. At the end of the exposure segment, pDCB did not produce an increase of 8-oxodG levels in the kidney nuclear DNA in contrast to potassium bromate (KBrO3). Following NTA promotion, no neoplastic lesions occurred in rats given pDCB, although diethylnitrosamine carcinogenesis was enhanced. Thus, pDCB did not produce oxidative DNA damage in the rat kidney or effect initiation of kidney carcinogenesis. These data suggest that oxidative stress is not involved in pDCB-induced renal carcinogenesis. The alpha2u-globulin-mediated chronic nephropathy probably acts as a promoter, not an initiation of renal carcinogenesis. Accordingly, pDCB is assessed to have no cancer hazard to humans who are not susceptible to the alpha2u-globulin nephropathy. PMID- 10817669 TI - Assessment of decalcifying protocols for detection of specific RNA by non radioactive in situ hybridization in calcified tissues. AB - For the best performance of in situ analysis of specific RNA expression in calcified tissues, it is necessary to choose an appropriate protocol to decalcify the tissues. We evaluated the usefulness of various acid-based decalcifying reagents with reference to 28 S rRNA staining by in situ hybridization using a thymine-thymine dimerized oligonucleotide probe. The reagents evaluated were 10% nitric acid, 10% HCl, 5% formic acid, 5% trichloroacetic acid, Morse's solution, Plank-Rychlo's solution, and K-CX solution, all of which are commonly used to decalcify tissues, and their effects on retention of morphology and RNA were compared with EDTA-based solutions. When normal mouse mandible was used as a model tissue, well-preserved morphology of ameloblasts was obtained from sections decalcified with Morse's solution, 10% HCl, Plank-Rychlo's solution, and K-CX solution, and best retention of 28 S rRNA was obtained with 5% formic acid and Morse's solution. We recommend Morse's solution to decalcify tissues to be processed for the rapid analysis of specific RNA expression. Indeed, we detected specific mRNAs strongly in sections treated with Morse's solution, and quantitative analysis showed that the ratio of signal intensities of 28 S rRNA and the specific mRNAs correlated with each other depending on decalcifying solutions. PMID- 10817670 TI - Improved quantitative characterization of atherosclerotic plaque composition with immunohistochemistry, confocal fluorescence microscopy, and computer-assisted image analysis. AB - To quantitatively characterize contributions of major constituents to the composition of a given atherosclerotic plaque, we have developed an approach employing immunohistochemistry, confocal scanning laser microscopy, and computer assisted image analysis. The method developed permits identification of plaques that are particularly vulnerable to rupture and elucidation of the nature of the composition of a given plaque, as well as the extent of luminal encroachment. Thus, it should be useful in experimental animals and ultimately in patients in delineating compositional changes in response to potentially deleterious genetic and environmentally induced factors and to potentially therapeutic interventions designed to diminish plaque vulnerability. PMID- 10817671 TI - Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization. AB - A method is presented to conjugate horseradish peroxidase (HRP) to oligodeoxynucleotides for fluorescence in situ hybridization assays employing tyramide signal amplification (TSA). HRP is covalently bound to the oligonucleotide by thiol ether linkage and purified by high-performance liquid chromatography. With TSA detection, a single HRP-labeled oligonucleotide probe is sufficient for in situ detection of clustered DNA repeat sequences with a degree of repetition between 20 and 50. PMID- 10817672 TI - Ultrastructural analysis of nucleolar transcription in cells microinjected with 5 bromo-UTP. AB - In situ sites of nucleolar transcription in cells microinjected with 5-bromo-UTP (BrUTP) were visualized at an ultrastructural level. After injection the cells were maintained for 4-90 min at 37 degrees C, fixed, and embedded in LR White resin. Postembedding immunoelectron microscopic visualization with colloidal gold has been used for localizing both Br-labeled precursor incorporated into pre-rRNA and different nucleolar transcription or processing factors. This high resolution approach allowed us to identify significant signal as early as after 4-min incubation periods following BrUTP microinjection. It revealed the dense fibrillar component (DFC) as being the first nucleolar compartment labeled with anti-bromodeoxyuridine antibody. Moreover, RNA polymerase I, nucleolar transcription factor UBF, and fibrillarin were also detected almost exclusively in this same nucleolar compartment. From 30 min onward, following microinjection, Br-labeled rRNA occurred also in the granular component. The results indicate that the DFC is the site of pre-rRNA transcription and of initial steps of pre rRNA processing. Moreover, it demonstrates that BrUTP microinjection followed by postembedding detection of Br-labeled RNA is a useful technique for high resolution studies of structure-function associations in the nucleolus. PMID- 10817673 TI - Expression of the apoptosis-inducing ligands FasL and TRAIL in malignant and benign human breast tumors. AB - Apoptosis-inducing ligands such as Fas ligand (FasL) and tumor necrosis factor related apoptosis-inducing ligand (TRAIL) have been found to play an important role in cell regulation. Different malignant tumors show an altered expression of these ligands and their respective receptors compared to normal tissues. The purpose of this study was therefore to investigate expression of TRAIL, FasL, and its receptor Fas on protein and mRNA levels in breast carcinomas (n=40), fibroadenomas (n=7), and normal breast tissues (n=5). Immunohistochemical reaction demonstrated that FasL was strongly expressed in breast cancer tissues (34/40) while only one fibroadenoma and one normal breast tissue reacted weakly positive for FasL. All fibroadenomas and normal breast tissues as well as the majority of breast cancer tissues expressed Fas on protein level. Quantitative RT PCR analysis detected high expression of FasL mRNA in breast cancer tissues and fibroadenomas, whereas fibroadenomas showed the highest Fas mRNA copy numbers, followed by breast cancer tissues and normal breast tissues (P<0.05). Compared to FasL expression, TRAIL could be detected in less breast cancer tissues on protein level (21/40) and was found in only one fibroadenoma and none of the normal breast tissues. Thus, it can be concluded that malignant breast tumors show an altered expression of the two apoptosis-inducing ligands FasL and TRAIL. PMID- 10817674 TI - Three-dimensional analysis of DNA replication foci: a comparative study on species and cell type in situ. AB - Chromatin morphology of interphase nuclei in most cell lines of quail (Coturnix coturnix japonica) and chick (Gallus gallus domesticus) embryos shows typical interspecies differences. This intrinsic marker has been used in quail/chick chimerisation experiments, where also differences between cell types were noted. We asked whether similar differences between species and between cell types could be observed in S phase nuclei in situ. In this report, we used bromodeoxyuridine (BrdU) pulse labelling and anti-BrdU immunofluorescence to detect DNA replication foci in the nuclei of identified cells. In the central nervous system of 5- to 7 day-old quail and chick embryos, mesoderm-derived cells with strikingly different morphology and topographical distribution were studied: endothelial, i.e. polarised cells forming continuous tubes, and macrophages, i.e. non-polarised, ameboid or ramified individual cells. Using confocal microscopy, replication foci in the nuclei were assessed quantitatively and three-dimensional visualisations were produced. We consistently observed that: (1) chick, but never quail, nuclei displayed completely confluent replication sites, independent of cell type, and (2) macrophages, but not endothelial cells, had distinct perinucleolar replication sites, independent of species. We thus demonstrate a new relationship between cell type and spatial arrangement of DNA replication sites, and conclude that interspecies differences of chromatin distribution are conserved throughout S phase. Our results strongly recommend that work done on nuclear structure in vitro should not be extrapolated without reservation to cells in vivo. PMID- 10817675 TI - Cytochemical demonstration of modification of carbohydrates in the mouse zona pellucida during folliculogenesis. AB - In the present study, lectin-gold cytochemistry and antibodies against ZP2 and ZP3 glycoproteins were used to investigate the oligosaccharide content of mouse ovarian zona pellucida (ZP) during follicular development. The entire thickness of the ZP and several organelles of the oocyte (cortical granules, Golgi apparatus, and vesicular aggregates) were reactive to RCA-I, DSA, AAA, WGA, MAA, and LFA throughout follicular development. HPA labeling was not detected at the earliest stages of follicular folliculogenesis. HPA reactivity was first observed in the ZP, Golgi apparatus, and the vesicles of oocytes at the trilaminar primary follicle stage. HPA labeling in the ZP was always restricted to the inner region of the zona matrix. After neuraminidase treatment, HPA reacted with the entire ZP in ovarian follicles at different stages of development. Immunolabeling with specific antibodies showed that, although ZP2 and ZP3 glycoproteins were uniformly distributed in the zona matrix of ovarian oocytes, there was a progressive increase in thickness of the ZP in parallel with the proliferation of follicular cells. ZP3 glycoprotein was also localized to the Golgi apparatus and vesicular aggregate. The present results suggest: (1) a difference in composition of carbohydrate content between the inner and outer region of the fully developed ZP generated probably by a modification in the biosynthetic pathway of oligosaccharides in the oocyte during folliculogenesis, (2) that newly synthesized ZP glycoproteins displace previously synthesized ZP components in a direction toward the follicular cells and, therefore, no redistribution of the ZP matrix occurs during folliculogenesis, and (3) that the vesicular aggregates in the ooplasm constitute an intermediate step in the secretory pathway of ZP glycoproteins. PMID- 10817676 TI - Characterization of caveolins from human knee joint catilage: expression of caveolin-1, -2, and -3 in chondrocytes and association with integrin beta1. AB - Interactions between the extracellular matrix (ECM) and chondrocytes are of great importance for structure and function of cartilage. The present study was undertaken to answer the question whether caveolins take part in integrin mediated cell-ECM interactions in the human cartilage. In samples of human knee joint cartilage, we detected the caveolin subtypes -1, -2, and -3 by immunohistochemical methods. Double-label experiments revealed a colocalization of caveolin with beta1-integrin. Results of immunoprecipitation and immunoblotting assays show that beta1-integrins associate with all three caveolin subtypes in human chondrocytes and indicate that they are part of the same complexes. Furthermore, immunoelectron microscopy shows the localization of beta1 integrin in caveolae-like structures of the cell membrane. The data stimulate further investigations on the role of the caveolin-integrin complex for integrin mediated signaling pathways in chondrocytes. PMID- 10817677 TI - Ultrastructural immunolocalization of basic fibroblast growth factor in fibroblasts and extracellular matrix. AB - Basic fibroblast growth factor (bFGF) is thought to play an important role in normal tissue repair and wound healing. It is a potent mitogenic and chemotactic factor for fibroblasts, regulating proliferation and extracellular matrix (ECM) production by these cells. In this study, we present morphologic evidence of the ultrastructural location of bFGF in fibroblasts and ECM using several antibodies, tissues, and species. Distinct labeling is seen in the nuclei of fibroblasts and some labeling in the cytosol. Immunolabeling of the cytosol excludes organelles involved in the usual secretory pathway, such as rough endoplasmic reticulum, Golgi apparatus, and secretory vacuoles. The same labeling is observed with either polyclonal or monoclonal antibodies. We suggest that bFGF functions as a nuclear protein in fibroblasts and is not secreted by a normal secretory pathway. Fibroblasts may export bFGF via unique cellular pathways that are clearly distinct from classic signal peptide mediated secretion. This may provide a source for ECM-resident bFGF. The same antibodies show different labeling intensity in the ECM. This protein, through integration into the ECM, may act as a local regulator and promote regeneration of these tissues after wounding. Direct evidence is the dramatic reduction of bFGF labeling in axotomized rat ECM collagen fibers versus control animals. PMID- 10817678 TI - Ultrastructural analysis of spinal primary afferent fibers within the circular muscle of the cat lower esophageal sphincter. AB - We have analyzed the ultrastructural characteristics and environment of spinal primary afferent fibers that run within the circular muscle of the cat lower esophageal sphincter. These were selectively labeled by anterogradely transported cholera toxin B subunit conjugated with horseradish peroxidase. Most of the labeled fibers were perpendicular to the muscle cells but some ran sinuously or parallel to the muscle cells. All the labeled fibers were unmyelinated and exhibited relatively rare varicosities. Most of the fibers were in large nerve fiber bundles surrounded by perineurium and probably project to the mucosa. Only some fibers that were in small nerve fiber bundles with no perineurium ran parallel to the musculature and established close relationships with smooth muscle cells. They might be a small subpopulation of the spinal tension receptors, most of the other spinal tension receptors being located in the myenteric plexus area, between the circular and longitudinal muscle. PMID- 10817679 TI - Immunohistochemical distribution of galectin-1, galectin-3, and olfactory marker protein in human olfactory epithelium. AB - The expression pattern of galectin-1 and galectin-3 in the human olfactory epithelium was investigated in relation to olfactory marker protein (OMP) using confocal laser immunofluorescence in human specimens and postmortem biopsies. OMP expression was found in olfactory receptor neurons (ORNs) in the olfactory mucosa and in fibers of the olfactory nerve crossing the submucous connective tissue. Galectin-1 was expressed in both the connective tissue of the nasal cavity and in the basal layer of the olfactory epithelium. In contrast, galectin-3 expression was limited to cells of the upper one-third of the olfactory epithelium. Expression of galectin-3 occurred in a subset of OMP-positive cells. However, between areas of galectin-1 and galectin-3 expression in the lower and upper portion of the epithelium, OMP-positive ORNs did not stain for both galectins. Considering the potential role of galectin-1 and galectin-3 in cell differentiation and maturation, the differential localization of galectins in the olfactory epithelium appears to be consistent with a significant role of these molecules in the physiological turnover of ORNs. PMID- 10817680 TI - 1999 Mutation Research Award for Scientific Excellence. Dr. John Essigmann. PMID- 10817681 TI - Mechanisms of morbidity and mortality from exposure to ambient air particles. AB - The studies reported here assessed pathophysiologic mechanisms that result from exposure to concentrated ambient particles (CAPs) in animals with and without cardiopulmonary compromise. These studies were carried out to determine the biologic plausibility of epidemiologic observations of increases in particulate air pollution associated with increases in human morbidity and mortality. Dogs were exposed two at a time to CAPs or filtered air via tracheostomy for six hours per day on three consecutive days. The electrocardiogram (ECG) and breathing pattern were recorded continuously, and indicators of inflammation were also assessed. In one experimental design, normal dogs were exposed in pairs to CAPs and subsequently to filtered air or to filtered air and subsequently CAPs (the double CAPs/double sham design). Comparisons were made between the CAPs measurements and each dog's own sham responses. In another design, one dog was exposed to CAPs while the chambermate received a sham exposure; these experiments were followed by crossover of the protocol the subsequent week (the crossover design). Comparisons were made between the CAPs exposure and both the chambermate's sham and each dog's own sham responses. The crossover experiments were conducted in normal animals and in animals who had undergone balloon occlusion of the left anterior descending (LAD) coronary artery to induce myocardial compromise. The effects of CAPs in animals with induced chronic bronchitis were part of the original specific aims; because these studies were not fully pursued, the results are presented only in Appendix A. In normal dogs, analyses of all double CAPs and crossover studies revealed low frequency (LF) and high frequency (HF) powers for heart rate variability (HRV) that were significantly higher for CAPs exposure compared to sham exposure. Variation in day-to-day exposure concentrations, aerosol composition, and pathophysiologic responses were also found. The crossover design, continuous measures of aerosol mass, and biologic responses were incorporated in the development of a statistical model that allowed isolation of changes associated with CAPs from changes due to animal variations. Comparison of individual exposures with this model revealed a range from no response in any measured parameter to statistically significant changes in cardiac autonomic balance, pulmonary air flow, and breathing pattern. On days in which dogs showed statistically significant changes in responses, the findings were consistent in both cardiac and respiratory parameters. Days associated with significant increases in LF and HF HRV, LF/ HF HRV ratio, and heart rate standard deviation (HR SD) were also associated with decreases in average heart rate. These same days had decreases in respiratory frequency, tidal volume, minute volume, and peak flows with corresponding increases in respiratory cycle times and enhanced pause (Pauenh), a measure of bronchoconstriction. These cardiac and respiratory changes suggest an effect mediated via both the sympathetic nervous system and the vagus nerve. Alternatively, days associated with increased heart rate had decreases in the HR SD; decreases or no change in HF and LF HRV; increases in respiratory flows and volumes; and decreases in breathing cycle times, all suggesting only sympathetic nervous system mediation. When all data from the crossover design experiments were assessed with this model, the heart rate and respiratory rate were significantly decreased in relation to both cumulative and actual exposure and the LF HRV, LF/HF HRV ratio, HR SD, and all other respiratory parameters were significantly increased (p < 0.0001 for all). When cardiac data were grouped by days in which the air mass trajectory came from the north or northwest (versus west, south, east, or northeast), significant increases in HR SD and HF HRV and significant decreases in average heart rate were associated with the northwest trajectory. (ABSTRACT TRUNCATED) PMID- 10817682 TI - Development and repair of cataract induced by ultraviolet radiation. AB - BACKGROUND: Several epidemiological investigations show a correlation between cataract development and the dose of ultraviolet radiation (UVR) received. It is experimentally well established that exposure of animal eyes to UVR induces cataract. Most cataracts develop as a gradual increase in lens opacity. Despite this, current estimations of toxicity for cataract are based on the concept that cataract is a binary event. Moreover, current exposure limits for UVR are based on subjective inspections with slit lamp microscopy. PURPOSE: The first purpose of the present study was to determine a statistically defined maximum acceptable dose for ultraviolet radiation-induced cataract based on quantitative data of forward light scattering in lenses. The second purpose was to find possible explanations for light scattering by investigating the morphology and the refractive index distribution in the lens. The third purpose was to describe the development of cataract after UVR on the cellular level. METHODS: Six-week-old, female Sprague-Dawley rats received UVR unilaterally in vivo. The radiation from a high pressure mercury lamp was collimated, passed through a water filter and an interference filter or a monochromator (lambdaMAX = 300 nm), and projected onto the cornea. The exposure time was 15 min. The exposure dose ranged between 0.1 and 20 kJ/m2 and the animals were kept between 6 hours and 32 weeks after exposure. The extracted lenses were photographed and forward light scattering was measured. Other methods included light microscopy, fluorescence microscopy, transmission and scanning electron microscopy, freeze-fracture and microradiography. RESULTS AND CONCLUSIONS: From a long-term experiment, it was concluded that UVR-exposed lenses scatter light more than their contralaterals and that a higher dose induces more light scattering. After exposure to 5 kJ/m2, the mean forward light scattering remains unchanged between 1 and 32 weeks. Earlier observations, taken together with the current findings, indicate that the optimal time to detect low dose UVR-induced cataract is one week after exposure in rats. The intensity of forward light scattering increases exponentially with increased UVR dose between 0.1 and 14 kJ/m2. Based on this continuous dose response, a method to determine a maximum acceptable dose to avoid UVR-induced cataract was developed. The statistically defined lower limit of pathologic light scattering is projected on the dose-response function. The dose corresponding to that point can be estimated and was suggested to be called the Maximum Acceptable Dose (MAD). Two low dose UVR exposures with 0 or 6 h intervals between the exposures produce the same degree of lens opacification. When the second exposure follows 24 or 48 h after the termination of the first, lenticular damage increases. Repair processes between 24 and 48 h after exposure appear to be sensitive to UVR, and an additional exposure during this time may aggravate cataract development. Lenses exposed to UVR grow more slowly than their non exposed contralaterals. This decrease in lens growth was more pronounced with increasing dose. Low doses led to decreased water content in the lens whereas high doses led to swelling. At 6 months after low dose UVR exposure, no global change of the refractive index was found. However, local variations of the refractive index induce a subtle cortical light scattering. In vivo low dose UVR induces programmed cell death which peaks 24 h post-exposure and involves the entire lens epithelium. Dead cells are removed from the epithelium by phagocytosis. This leads to disintegration of the lens epithelium, associated with flake-like opacities at the lens surface. After one week, the epithelium and the equatorial parts of superficial lens fibers contain extracellular spaces. The extracellular spaces together with locally disarranged fibers produce a corrugated opaque lens surface and equatorial opacities. Within several weeks after ex PMID- 10817685 TI - Quinupristin/Dalfopristin therapy for infections due to vancomycin-resistant Enterococcus faecium. AB - The efficacy and safety of quinupristin/dalfopristin for treatment of infections due to vancomycin-resistant Enterococcus faecium were evaluated in 24 hospitalized patients with documented infections (19 bacteremias, 5 localized infections) caused by vancomycin-resistant E. faecium that was susceptible to quinupristin/dalfopristin in vitro. Patients received iv quinupristin/dalfopristin at a dosage of either 7.5 mg/kg every 8 h or 5 mg/kg every 8 h. A favorable clinical response (cure or improvement) occurred in 19 (83%) of 23 evaluable patients; bacteriologic eradication occurred in 17 (74%) of 23 evaluable patients. A favorable clinical response was observed in 12 (80%) of 15 patients who were treated with 7.5 mg/kg of quinupristin/dalfopristin every 8 h and in 7 (88%) of 8 patients treated with 5 mg/kg of quinupristin/dalfopristin every 8 h. Two of four treatment failures were associated with a decrease in the in vitro susceptibility of vancomycin-resistant E. faecium to quinupristin/dalfopristin. Superinfections developed in 6 patients (26%), but only one was caused by Enterococcus faecalis that was resistant to quinupristin/dalfopristin. Myalgias and arthralgias were the only adverse events related to quinupristin/dalfopristin. These conditions occurred in 8 (33%) of 24 patients and were dose-related (8 cases in 16 patients treated with 7.5 mg/kg of quinupristin/dalfopristin every 8 h, no cases in 8 patients treated with 5 mg/kg every 8 h). Mortality associated with vancomycin-resistant E. faecium infection was 17% (4 of 23 patients), whereas mortality from other causes was 52% (12 of 23 patients). These results suggest that quinupristin/dalfopristin is effective as treatment for vancomycin-resistant E. faecium infections in critically ill patients with serious underlying conditions. Except for myalgias and arthralgias at higher dosages, the drug is well-tolerated. PMID- 10817686 TI - Evernimicin (SCH27899) inhibits both translation and 50S ribosomal subunit formation in Staphylococcus aureus cells. AB - The effects of the everninomicin antibiotic evernimicin (SCH27899) on growing Staphylococcus aureus cells were investigated. Cellular growth rates and viable cell numbers decreased with increasing antibiotic concentrations. The rate of protein synthesis, measured as (35)S-amino acid incorporation, declined in parallel with the growth rate. Significantly, the formation of the 50S ribosomal subunit was inhibited in a dose-dependent fashion as well. 30S ribosomal subunit synthesis was not affected over the same concentration range. Evernimicin did not stimulate the breakdown of mature ribosomal subunits. Pulse-chase labeling experiments revealed a reduced rate of 50S subunit formation in drug-treated cells. Two erythromycin-resistant strains of S. aureus that carried the ermC gene were as sensitive as wild-type cells to antibiotic inhibition. In addition, two methicillin-resistant S. aureus organisms, one sensitive to erythromycin and one resistant to the macrolide, showed similar sensitivities to evernimicin. These results suggest a use for this novel antimicrobial agent against antibiotic resistant bacterial infections. PMID- 10817687 TI - Identification and analysis of bacterial protein secretion inhibitors utilizing a SecA-LacZ reporter fusion system. AB - Protein secretion is an essential process for bacterial growth, yet there are few if any antimicrobial agents which inhibit secretion. An in vivo, high-throughput screen to detect secretion inhibitors was developed based on the translational autoregulation of one of the central protein components, SecA. The assay makes use of a SecA-LacZ fusion reporter construct in Escherichia coli which is induced when secretion is perturbed. Several compounds, including two natural product extracts, which had the ability to induce the reporter fusion were identified and the MICs of these compounds for Staphylococcus aureus strain MN8 were found to be < or =128 microg/ml. Enzyme-linked immunosorbent assay, Western blotting, and immunoprecipitation techniques were used to analyze the affects of these compounds on protein secretion. Six representative compounds presented here appear to be bona fide secretion inhibitors but were found to have deleterious effects on membranes. It was concluded that, while the method described here for identifying inhibitors of secretion is valid, screens such as this, which are directed against the membrane-bound portion of a pathway, may preferentially identify compounds which affect membrane integrity. PMID- 10817689 TI - Prevalence of SHV-12 among clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamases and identification of a novel AmpC enzyme (CMY 8) in Southern Taiwan. AB - Twenty (8.5%) of 234 nonrepetitive clinical isolates of Klebsiella pneumoniae from southern Taiwan were found to produce extended-spectrum beta-lactamases (ESBLs): 10 strains produced SHV-12, 4 produced SHV-5, 2 produced a non-TEM non SHV ESBL with a pI of 8.3, 3 produced a novel AmpC beta-lactamase designated CMY 8 with a pI of 8.25, and 1 produced SHV-12 and an unidentified AmpC enzyme with a pI of 8.2. The CMY-8 enzyme confers a resistance phenotype similar to CMY-1 and MOX-1, and sequence comparisons showed high homologies (>95%) of nucleotide and amino acid sequences among these three enzymes. Plasmid and pulse-field gel electrophoresis analyses revealed that all isolates harboring an SHV-derived ESBL were genetically unrelated, indicating that dissemination of resistance plasmids is responsible for the spread of SHV ESBLs among K. pneumoniae in this area. All three isolates carrying CMY-8 had identical genotypic patterns, suggesting the presence of an epidemic strain. PMID- 10817688 TI - Induction of fibronectin-binding proteins and increased adhesion of quinolone resistant Staphylococcus aureus by subinhibitory levels of ciprofloxacin. AB - We recently reported that strain EN1252a, a fluoroquinolone-resistant derivative of Staphylococcus aureus NCTC8325 with mutations in grlA and gyrA, expressed increased levels of fibronectin-binding proteins (FnBPs) and showed a significantly higher attachment to fibronectin-coated polymer surfaces after growth in the presence of subinhibitory concentrations of ciprofloxacin. The present study evaluated the occurrence and frequency of fluoroquinolone-induced FnBP-mediated adhesion in clinical isolates of fluoroquinolone-resistant methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA). Eight of ten MRSA isolates and four of six MSSA isolates with grlA and gyrA mutations exhibited significant increases in attachment to fibronectin coated surfaces after growth in the presence of one-quarter the MIC of ciprofloxacin. Fluoroquinolone-induced FnBP-mediated adhesion of one clinical MRSA strain and the double mutant strain EN1252a also occurred on coverslips removed from the subcutaneous space of guinea pigs. For strain EN1252a, the regulation of fnb transcription by sub-MICs of ciprofloxacin was studied on reporter plasmids carrying fnb-luxAB fusions. One-quarter of the MIC of ciprofloxacin significantly increased fnbB, but not fnbA, promoter activity of the fluoroquinolone-resistant mutant but not its fluoroquinolone-susceptible parent ISP794. This response was abolished by pretreatment with rifampin, indicating an effect at the level of transcription. Activation of the fnbB promoter was not due to an indirect effect of ciprofloxacin on growth rate and still occurred in an agr mutant of strain EN1252a. These data suggest that sub MIC levels of ciprofloxacin activate the fnbB promoter of some laboratory and clinical isolates, thus contributing to increased production of FnBP(s) and leading to higher levels of bacterial attachment to fibronectin-coated or subcutaneously implanted coverslips. PMID- 10817690 TI - Pharmacokinetics of gentamicin C(1), C(1a), and C(2) in beagles after a single intravenous dose. AB - The pharmacokinetics of gentamicin C(1), C(2), and C(1a) were studied in six beagles after administration of gentamicin at 4 mg/kg of body weight as a single intravenous bolus dose. Plasma concentrations of the gentamicin components were analyzed with a novel high-performance liquid chromatography method capable of identifying and quantifying each of the components. The pharmacokinetic analysis of the plasma concentration-versus-time data was performed using the noncompartmental approach. The results indicated significant differences in the pharmacokinetic characteristics between the gentamicin components C(1), C(1a), and C(2). The mean residence times of gentamicin C(1), C(1a), and C(2) were 81+/ 13, 84+/-12, and 79+/-13 min (mean +/- standard deviation), respectively. The half-lives of the respective components were 64+/-12, 66+/-12 and 63+/-12 min. Clearance (CL) of gentamicin C(1), 4.62+/-0.71 ml min(-1) kg(-1), was significantly higher (P = 0.0156) than CL of gentamicin C(1a), 1.81+/-0.26 ml min(-1) kg(-1), and C(2), 1.82+/-0.25 ml min(-1) kg(-1). Similarly, the volume of distribution at steady state (V(ss)) of gentamicin C(1), 0.36+/-0.04 liter kg( 1), was significantly higher (P = 0.0156) than the V(ss) of gentamicin C(1a), 0.14+/-0.01 liter kg(-1), and C(2), 0.15+/-0.02 liter kg(-1). Tissue binding was considered the most likely cause for the difference. The difference may have clinical and toxicological significance. PMID- 10817691 TI - Carbapenemases of Chryseobacterium (Flavobacterium) meningosepticum: distribution of blaB and characterization of a novel metallo-beta-lactamase gene, blaB3, in the type strain, NCTC 10016. AB - Genes encoding carbapenemases in 15 reference strains of Chryseobacterium (Flavobacterium) meningosepticum from the United Kingdom National Collection of Type Cultures and in one recent clinical isolate were investigated. All the strains hydrolyzed imipenem, but their levels of resistance to carbapenems varied, with imipenem and meropenem MICs ranging from 2 to >32 microg/ml. The blaB gene, which encodes a molecular-class B carbapenemase, was detected in only six reference strains and in clinical isolate 97/P/5448. The gene from 97/P/5448 had 98% nucleotide identity with the published sequence of blaB (from strain NCTC 10585) and was designated blaB2. A distinct carbapenemase gene, designated blaB3, was cloned from the type strain of C. meningosepticum, NCTC 10016. blaB3 had an open reading frame of 750 bp with 82% nucleotide identity to blaB and blaB2 and encoded a beta-lactamase of 249 amino acids, including the putative signal peptide. This beta-lactamase showed 87.6 and 86.7% amino acid homology with BlaB and BlaB2, respectively. blaB3 was detected in one other reference strain besides NCTC 10016, but the genetic basis of the carbapenemase activity detected in the other seven reference strains was not defined. Thus, neither blaB nor blaB3 was ubiquitous in the strains of C. meningosepticum studied, indicating that the reference strains may represent more than one bacterial species, each with its own intrinsic metallo-beta-lactamase. Further taxonomic studies of C. meningosepticum are necessary to resolve this topic. Chryseobacterium spp. are environmental organisms and occasional opportunist pathogens. They apparently represent a reservoir of diverse metallo-beta-lactamases, which potentially spread to gram-negative bacteria of greater clinical significance. PMID- 10817692 TI - Resistance to macrolides in Streptococcus pyogenes in France in pediatric patients. AB - A total of 1,500 recent throat isolates of Streptococcus pyogenes collected between 1996 and 1999 from children throughout France were tested for their susceptibility to erythromycin, azithromycin, josamycin, clindamycin, and streptogramin B. The erythromycin-resistant isolates were further studied for their genetic mechanism of resistance, by means of PCR. The clonality of these strains was also investigated by means of serotyping and ribotyping. In all, 6.2% of the strains were erythromycin resistant, and 3.4 and 2.8% expressed the constitutive MLS(B) and M resistance phenotypes and harbored the ermB and mefA genes, respectively; ermTR was recovered from one isolate which also harbored the ermB gene. Ten serotypes and 8 ribotypes were identified, but we identified 17 strains by combining serotyping with ribotyping. Among the eight ribotypes, the mefA gene was recovered from six clusters, one being predominant, while the ermB gene was recovered from four clusters, of which two were predominant. PMID- 10817693 TI - Evaluation of rifalazil in long-term treatment regimens for tuberculosis in mice. AB - Previous experiments with rifalazil (RLZ) (also known as KRM-1648) in combination with isoniazid (INH) demonstrated its potential for short-course treatment of Mycobacterium tuberculosis infection. In this study we investigated the minimum RLZ-INH treatment time required to eradicate M. tuberculosis in a murine model. RLZ-INH treatment for 6 weeks or longer led to a nonculturable state. Groups of mice treated in parallel were killed following an observation period to evaluate regrowth. RLZ-INH treatment for a minimum of 10 weeks was necessary to maintain a nonculturable state through the observation period. Pyrazinamide (PZA) was added to this regimen to determine whether the treatment duration could be further reduced. In this model, the addition of PZA did not shorten the duration of RLZ INH treatment required to eradicate M. tuberculosis from mice. The addition of PZA reduced the number of mice in which regrowth occurred, although the reduction was not statistically significant. PMID- 10817694 TI - Antifungal activity of amphotericin B cochleates against Candida albicans infection in a mouse model. AB - Cochleates are lipid-based supramolecular assemblies composed of natural products, negatively charged phospholipid, and a divalent cation. Cochleates can encapsulate amphotericin B (AmB), an important antifungal drug. AmB cochleates (CAMB) have a unique shape and the ability to target AmB to fungi. The minimal inhibitory concentration and the minimum lethal concentration against Candida albicans are similar to that for desoxycholate AmB (DAMB; Fungizone). In vitro, CAMB induced no hemolysis of human red blood cells at concentrations of as high as 500 microg of AmB/ml, and DAMB was highly hemolytic at 10 microg of AmB/ml. CAMB protect ICR mice infected with C. albicans when the agent is administered intraperitoneally at doses of as low as 0.1 mg/kg/day. In a tissue burden study, CAMB, DAMB, and AmBisome (liposomal AmB; LAMB) were effective in the kidneys, but in the spleen CAMB was more potent than DAMB at 1 mg/kg/day and was equivalent to LAMB at 10 mg/kg/day. In summary, CAMB are highly effective in treating murine candidiasis and compare well with AmBisome and AmB. PMID- 10817695 TI - Biochemical-genetic characterization and regulation of expression of an ACC-1 like chromosome-borne cephalosporinase from Hafnia alvei. AB - A naturally occurring AmpC beta-lactamase (cephalosporinase) gene was cloned from the Hafnia alvei 1 clinical isolate and expressed in Escherichia coli. The deduced AmpC beta-lactamase (ACC-2) had a pI of 8 and a relative molecular mass of 37 kDa and showed 50 and 47% amino acid identity with the chromosome-encoded AmpCs from Serratia marcescens and Providentia stuartii, respectively. It had 94% amino acid identity with the recently described plasmid-borne cephalosporinase ACC-1 from Klebsiella pneumoniae, suggesting the chromosomal origin of ACC-1. The hydrolysis constants (k(cat) and K(m)) showed that ACC-2 was a peculiar cephalosporinase, since it significantly hydrolyzed cefpirome. Once its gene was cloned and expressed in E. coli (pDEL-1), ACC-2 conferred resistance to ceftazidime and cefotaxime but also an uncommon reduced susceptibility to cefpirome. A divergently transcribed ampR gene with an overlapping promoter compared with ampC (bla(ACC-2)) was identified in H. alvei 1, encoding an AmpR protein that shared 64% amino acid identity with the closest AmpR protein from P. stuartii. beta-Lactamase induction experiments showed that the ampC gene was repressed in the absence of ampR and was activated when cefoxitin or imipenem was added as an inducer. From H. alvei 1 cultures that expressed an inducible cephalosporinase phenotype, several ceftazidime- and cefpirome-cross-resistant H. alvei 1 mutants were obtained upon selection on cefpirome- or ceftazidime containing plates, and H. alvei 1 DER, a ceftazidime-resistant mutant, stably overproduced cephalosporinase. Transformation of H. alvei 1 DER or E. coli JRG582 (ampDE mutant) harboring ampC and ampR from H. alvei 1 with a recombinant plasmid containing ampD from E. coli resulted in a decrease in the MIC of beta-lactam and recovery of an inducible phenotype for H. alvei 1 DER. Thus, AmpR and AmpD proteins may regulate biosynthesis of the H. alvei cephalosporinase similarly to other enterobacterial cephalosporinases. PMID- 10817696 TI - A between-species comparison of antimicrobial resistance in enterobacteria in fecal flora. AB - Enterobacteria in fecal flora are often reported to be highly resistant. Escherichia coli is the main species; resistance data on other species are rare. To assess the effect of the host's environment, antimicrobial resistance was determined in fecal species of the family Enterobacteriaceae from three populations: healthy people (HP)(n = 125) with no exposure to antimicrobials for 3 months preceding sampling, university hospital patients (UP) (n = 159) from wards where the antibiotic use was 112 defined daily doses (DDD)/bed/month, and geriatric long-term patients (LTP) (n = 74) who used 1.8 DDD/bed/month. The mean length of hospital stay was 5 days for the UP and 22 months for the LTP. The isolates were identified to at least genus level, and MICs of 16 antimicrobials were determined. From the university hospital, resistance data on clinical Enterobacteriaceae isolates were also collected. Resistance data for on average two different isolates per sample (range, 1 to 5) were analyzed: 471 E. coli isolates and 261 other Enterobacteriaceae spp. Resistance was mainly found among E. coli; even in HP, 18% of E. coli isolates were resistant to two or more antimicrobial groups, with MIC patterns indicative of transferable resistance. Other fecal enterobacteria were generally susceptible, with little typically transferable multiresistance. Clinical Klebsiella and Enterobacter isolates were significantly more resistant than fecal isolates. The resistance patterns at both hospitals mirrored the patterns of antibiotic use, but LTP E. coli isolates were significantly more resistant than those from UP. Conditions permitting an efficient spread may have been more important in sustaining high resistance levels in the LTP. E. coli was the main carrier of antimicrobial resistance in fecal flora; resistance in other species was rare in the absence of antimicrobial selection. PMID- 10817697 TI - In vitro assessment of antifungal therapeutic potential of salivary histatin-5, two variants of histatin-5, and salivary mucin (MUC7) domain 1. AB - Human salivary histatin-5 (Hsn-5) is a 24-residue peptide that possesses potent antifungal activity in vitro. The MUC7 gene encodes human salivary low-molecular weight mucin (MG2). The candidacidal activity of MUC7 domain 1 (MUC7 D1, the N terminal 51 amino acid residues of MUC7) in vitro has also been demonstrated. In this study, we have investigated the antifungal therapeutic potential of Hsn-5, its two variants, R12I/K17N and R12I/H21L, and MUC7 D1. First, these peptides were tested for activities against different clinically important fungi. We found them to possess broad-spectrum antifungal activities; specifically, most exhibited excellent in vitro activity against eight clinically important fungal strains tested, including Candida albicans and Candida glabrata and their azole resistant counterparts and Cryptococcus neoformans and its amphotericin B resistant counterpart. These findings also suggest that the mechanism of action of both Hsn-5 and MUC7 D1 for these fungi is different from that of amphotericin B or azole antifungal agents. Second, we examined the stability of these peptides in whole human saliva and human serum. In saliva, the Hsn-5 variants R12I/K17N and R12I/H21L and MUC7 D1 degraded at a lower rate than Hsn-5. In human serum, MUC7 D1 was also more stable than Hsn-5; both peptides were more stable in serum than in saliva. Third, we examined the cytotoxicity of these peptides using human erythrocytes and two human cell lines (KB and HSG). No (or very low) hemolytic activity was observed with any of the four peptides, even at the highest protein concentration tested (200 microM), while amphotericin B caused 100% hemolysis at only 12.5 microM. The toxic effects of Hsn-5 and MUC7 D1 toward KB and HSG cells were also much lower than that of amphotericin B as measured by trypan blue exclusion. Together, these findings indicate that the investigated peptides possess high antifungal therapeutic potential, in particular for the treatment of drug-resistant fungal strains associated with immunocompromised (particularly human immunodeficiency virus-infected) patients. The same peptides could also be used as components of artificial saliva for patients with salivary dysfunction. PMID- 10817698 TI - Activity of the novel immunomodulatory compound tucaresol against experimental visceral leishmaniasis. AB - Tucaresol, a novel immunomodulator, was inactive against Leishmania donovani amastigotes in both peritoneal and bone marrow macrophages in vitro at concentrations between 100 and 1 microM, with toxicity to macrophages and parasites at 300 microM. However, against L. donovani in BALB/c mice at doses between 80 and 1.25 mg/kg of body weight administered once daily by the oral route during days 7 to 11 of infection, an optimal dose of 5 mg/kg produced a 43.8 to 62.4% suppression of liver amastigotes, with significantly reduced activity at the extremes of the dose range. This response was not related to levels of infection. No interaction with the standard pentavalent antimonial sodium stibogluconate (Pentostam) was observed during this period of infection. The optimum dose of 5 mg/kg was ineffective when administered during the first week of infection and was most effective against the liver infection when administered during weeks 2 to 3 of infection (42.3 to 46.8% inhibition) and against the splenic infection when administered during week 6 of infection (59.5% inhibition). The optimum dose of tucaresol against L. donovani in C57BL/6 mice was 5 mg/kg, which produced a 40.8 to 48.7% suppression of liver amastigotes when administered in a range of 80 to 1.25 mg/kg during days 7 to 11 of infection. The drug had no activity against L. donovani infections in C.B-17 scid mice when the same regimen was used. PMID- 10817699 TI - A novel complex mutant beta-lactamase, TEM-68, identified in a Klebsiella pneumoniae isolate from an outbreak of extended-spectrum beta-lactamase-producing Klebsiellae. AB - Twenty-two Klebsiella pneumoniae and two K. oxytoca extended-spectrum beta lactamase (ESBL)-producing isolates were collected in 1996 from patients in two pediatric wards of the University Hospital in Wroclaw, Poland. Molecular typing has revealed that the K. pneumoniae isolates represented four different epidemic strains. Three kinds of enzymes with ESBL activity (pI values of 5.7, 6.0, and 8.2) were identified. The pI 6.0 beta-lactamases belonged to the TEM family, and sequencing of the bla(TEM) genes amplified from representative isolates revealed that these enzymes were TEM-47, previously identified in K. pneumoniae isolates from pediatric hospitals in Lodz and Warsaw. One of the TEM-47-producing strains from Wroclaw was very closely related to the isolates from the other cities, and this indicated countrywide spread of the epidemic strain. The pI 5.7 beta lactamase was produced by a single K. pneumoniae isolate for which, apart from oxyimino-beta-lactams, the MICs of beta-lactam-inhibitor combinations were also remarkably high. Sequencing revealed that this was a novel TEM beta-lactamase variant, TEM-68, specified by the following combination of mutations: Gly238Ser, Glu240Lys, Thr265Met, and Arg275Leu. The new enzyme has most probably evolved from TEM-47 by acquiring the single substitution of Arg275, which before was identified only twice in enzymes with inhibitor resistance (IR) activity. TEM-68 was shown to be a novel complex mutant TEM beta-lactamase (CMT-2) which combines strong ESBL activity with relatively weak IR activity and, when expressed in K. pneumoniae, is able to confer high-level resistance to a wide variety of beta lactams, including inhibitor combinations. This data confirms the role of the Arg275Leu mutation in determining IR activity and documents the first isolation of K. pneumoniae producing the complex mutant enzyme. PMID- 10817700 TI - In vitro activities of methylenecyclopropane analogues of nucleosides and their phosphoralaninate prodrugs against cytomegalovirus and other herpesvirus infections. AB - Human cytomegalovirus (HCMV) infection does not generally cause problems in the immunocompetent adult but can result in severe clinical disease in the fetus, neonate, and immunocompromised host. Ganciclovir (GCV), the agent currently used to treat most HCMV infections, has resulted in much therapeutic success; however, efficacy remains suboptimal. Therefore, there is still a need to develop new compounds for use against HCMV infections. In the present study, several Z- and E series methylenecyclopropane analogues and their phosphoroalaninate prodrugs were tested initially for activity against HCMV, strain AD169, and murine cytomegalovirus (MCMV) in vitro. Many were found to exhibit efficacy comparable to that of GCV against HCMV in plaque assays and were active against MCMV as well. The compounds were also tested for efficacy against herpes simplex virus types 1 and 2, varicella-zoster virus, and Epstein-Barr virus, and some had levels of activity that were comparable to that of acyclovir. In addition, the compounds synguanol (QYL-438) and 2-amino-6-cyclopropylamino analogue (QYL-769) were chosen for further evaluation and were found to be effective against additional laboratory and clinical isolates of HCMV and GCV-resistant isolates. QYL-438 and QYL-769 were found to be nontoxic in human and mouse fibroblasts and were considerably less toxic than GCV in granulocyte macrophage CFUs and erythroid burst-forming units. These results provide evidence for the high activity of some of these methylenecyclopropane analogues against various herpesviruses, particularly HCMV, in tissue culture and suggest that further evaluation is warranted to determine their potential for use in future clinical studies. PMID- 10817701 TI - Comparison of fluconazole and itraconazole in a rabbit model of coccidioidal meningitis. AB - Coccidioidal meningitis is a devastating disease that requires long-term therapy with little hope of cure. A rabbit model of coccidioidal meningitis was used to compare the therapeutic efficacies of fluconazole (FCZ) and itraconazole (ITZ). Hydrocortisone-treated male New Zealand white rabbits were infected intracisternally with 5.0x10(4) to 5.4x10(4) arthroconidia of Coccidioides immitis. Oral treatment with polyethylene glycol 200 (PEG) (n = 9), FCZ (n = 8; 80 mg/kg of body weight/day), or ITZ (n = 8; 80 mg/kg/day) began 5 days after infection and continued for 28 consecutive days. Both FCZ and ITZ reduced the number of CFU of C. immitis organisms in the spinal cord and brain compared with the number in PEG-treated animals (P< or =0.003), but the results for FCZ and ITZ were not different from each other. Histopathologic severity (semiquantitative scoring system by an observer blinded to treatment) was equally reduced in both FCZ and ITZ treatment groups compared with that in controls (P< or =0.0004). Both treatments resulted in lower cerebrospinal fluid (CSF) protein concentrations and leukocyte counts and faster clearing of C. immitis from CSF compared with the results for PEG-treated controls. Neither drug affected CSF glucose levels. Both compounds were effective at reducing neurological and systemic signs and extending survival (P< or =0.014). FCZ was more effective at reducing head and body shakes, posture changes, and incontinence; ITZ was more effective at reducing continuous fever. Mean levels of FCZ and ITZ in the serum and CSF were determined by bioassay; at 17 to 26 h postdosing, levels were 28.1 to 40.0 and 22.4 to 29.9 microg/ml, respectively, for FCZ and 0.77 to 2.51 and 0 microg/ml, respectively, for ITZ. The sera of most animals developed antibody to C. immitis, but azole treatment attenuated antibody development in CSF and its titer. In conclusion, both FCZ and ITZ were efficacious, but neither was curative in a rabbit model of coccidioidal meningitis. PMID- 10817702 TI - Morphological changes and lysis induced by beta-lactams associated with the characteristic profiles of affinities of penicillin-binding proteins in actinobacillus pleuropneumoniae. AB - Actinobacillus pleuropneumoniae, which was formerly classified in the genus Haemophilus, is a pathogen causing swine pleuropneumonia. We found that aspoxicillin showed strong activity and that meropenem had better lytic activity against this pathogen. In the present study, we for the first time identified penicillin-binding proteins (PBPs) of A. pleuropneumoniae in order to elucidate the relationship between the antibacterial and lytic activities of beta-lactam antibiotics and affinities of the PBPs. The competitive assay using (3)H-labeled benzylpenicillin revealed seven PBPs in A. pleuropneumoniae; they were determined to be PBPs 1a, 1b, 2, 3, 4, 5, and 6, and the molecular masses of these PBPs were estimated to be 92, 80, 76, 72, 50, 44, and 30 kDa, respectively, by comparison with those of Haemophilus influenzae. Our detailed analysis of the affinities of the PBPs of A. pleuropneumoniae and of the bacterial lysis kinetics for several beta-lactam antibiotics revealed that the strong antibacterial activity of aspoxicillin against this strain could be related to the higher affinity of PBP 3 and that preferential inactivation of PBP 1b could cause rapid lysis. PMID- 10817703 TI - Difference in incidence of spontaneous mutations between Herpes simplex virus types 1 and 2. AB - Spontaneous mutations within the herpes simplex virus (HSV) genome are introduced by errors during DNA replication. Indicative of the inherent mutation rate of HSV DNA replication, heterogeneous HSV populations containing both acyclovir (ACV) resistant and ACV-sensitive viruses occur naturally in both clinical isolates and laboratory stocks. Wild-type, laboratory-adapted HSV type 1 (HSV-1) strains KOS and Cl101 reportedly accumulate spontaneous ACV-resistant mutations at a frequency of approximately six to eight mutants per 10(4) plaque-forming viruses (U. B. Dasgupta and W. C. Summers, Proc. Natl. Acad. Sci. USA 75:2378-2381, 1978; J. D. Hall, D. M. Coen, B. L. Fisher, M. Weisslitz, S. Randall, R. E. Almy, P. T. Gelep, and P. A. Schaffer, Virology 132:26-37, 1984). Typically, these resistance mutations map to the thymidine kinase (TK) gene and render the virus TK deficient. To examine this process more closely, a plating efficiency assay was used to determine whether the frequencies of naturally occurring mutations in populations of the laboratory strains HSV-1 SC16, HSV-2 SB5, and HSV-2 333 grown in MRC-5 cells were similar when scored for resistance to penciclovir (PCV) and ACV. Our results indicate that (i) HSV mutants resistant to PCV and those resistant to ACV accumulate at approximately equal frequencies during replication in cell culture, (ii) the spontaneous mutation frequency for the HSV-1 strain SC16 is similar to that previously reported for HSV-1 laboratory strains KOS and Cl101, and (iii) spontaneous mutations in the laboratory HSV-2 strains examined were 9- to 16-fold more frequent than those in the HSV-1 strain SC16. These observations were confirmed and extended for a group of eight clinical isolates in which the HSV-2 mutation frequency was approximately 30 times higher than that for HSV-1 isolates. In conclusion, our results indicate that the frequencies of naturally occurring, or spontaneous, HSV mutants resistant to PCV and those resistant to ACV are similar. However, HSV-2 strains may have a greater propensity to generate drug-resistant mutants than do HSV-1 strains. PMID- 10817704 TI - Dihydropteroate synthase of Mycobacterium leprae and dapsone resistance. AB - Two Mycobacterium leprae genes, folP1 and folP2, encoding putative dihydropteroate synthases (DHPS), were studied for enzymatic activity and for the presence of mutations associated with dapsone resistance. Each gene was cloned and expressed in a folP knockout mutant of Escherichia coli (C600DeltafolP::Km(r)). Expression of M. leprae folP1 in C600DeltafolP::Km(r) conferred growth on a folate-deficient medium, and bacterial lysates exhibited DHPS activity. This recombinant displayed a 256-fold-greater sensitivity to dapsone (measured by the MIC) than wild-type E. coli C600, and 50-fold less dapsone was required to block (expressed as the 50% inhibitory concentration [IC(50)]) the DHPS activity of this recombinant. When the folP1 genes of several dapsone-resistant M. leprae clinical isolates were sequenced, two missense mutations were identified. One mutation occurred at codon 53, substituting an isoleucine for a threonine residue (T53I) in the DHPS-1, and a second mutation occurred in codon 55, substituting an arginine for a proline residue (P55R). Transformation of the C600DeltafolP::Km(r) knockout with plasmids carrying either the T53I or the P55R mutant allele did not substantially alter the DHPS activity compared to levels produced by recombinants containing wild-type M. leprae folP1. However, both mutations increased dapsone resistance, with P55R having the greatest affect on dapsone resistance by increasing the MIC 64-fold and the IC(50) 68-fold. These results prove that the folP1 of M. leprae encodes a functional DHPS and that mutations within this gene are associated with the development of dapsone resistance in clinical isolates of M. leprae. Transformants created with M. leprae folP2 did not confer growth on the C600DeltafolP::Km(r) knockout strain, and DNA sequences of folP2 from dapsone susceptible and -resistant M. leprae strains were identical, indicating that this gene does not encode a functional DHPS and is not involved in dapsone resistance in M. leprae. PMID- 10817705 TI - The Legionella (Fluoribacter) gormanii metallo-beta-lactamase: a new member of the highly divergent lineage of molecular-subclass B3 beta-lactamases. AB - A metallo-beta-lactamase determinant was cloned from a genomic library of Legionella (Fluoribacter) gormanii ATCC 33297(T) constructed in the plasmid vector pACYC184 and transformed into Escherichia coli DH5alpha, by screening for clones showing a reduced susceptibility to imipenem. The product of the cloned determinant, named FEZ-1, contains a 30-kDa polypeptide and exhibits an isoelectric pH of 7.6. Sequencing revealed that FEZ-1 is a molecular-class B beta lactamase which shares the closest structural similarity (29.7% of identical residues) with the L1 enzyme of Stenotrophomonas maltophilia, being a new member of the highly divergent subclass B3 lineage. All the residues that in L1 are known to be directly or indirectly involved in coordination of the zinc ions were found to be conserved also in FEZ-1, suggesting that the geometry of zinc coordination in the active site of the latter enzyme is identical to that of L1. Unlike L1, however, FEZ-1 appeared to be monomeric in gel permeation chromatography experiments and exhibited a distinctive substrate specificity with a marked preference for cephalosporins and meropenem. The properties of FEZ-1 overall resembled those of a beta-lactamase previously purified from the same strain of L. gormanii (T. Fujii, K. Sato, K. Miyata, M. Inoue, and S. Mitsuhashi, Antimicrob. Agents Chemother. 29:925-926, 1986) and are as yet unique among class B enzymes, reinforcing the notion that considerable functional heterogeneity can be encountered among members of this class. A system for overexpression of the bla(FEZ-1) gene in E. coli, based on the T7 phage promoter, was also developed. PMID- 10817706 TI - Correlation of fluconazole MICs with clinical outcome in cryptococcal infection. AB - We have correlated the in vitro results of testing the susceptibility of Cryptococcus neoformans to fluconazole with the clinical outcome after fluconazole maintenance therapy in patients with AIDS-associated cryptococcal disease. A total of 28 isolates of C. neoformans from 25 patients (24 AIDS patients) were tested. The MICs were determined by the broth microdilution technique by following the modified guidelines described in National Committee for Clinical Standards (NCCLS) document M27-A, e.g., use of yeast nitrogen base medium and a final inoculum of 10(4) CFU/ml. The fluconazole MIC at which 50% of isolates are inhibited (MIC(50)) and MIC(90), obtained spectrophotometrically after 48 h of incubation, were 4 and 16 microg/ml, respectively. Of the 25 patients studied, 4 died of active cryptococcal disease and 2 died of other causes. Therapeutic failure was observed in five patients who were infected with isolates for which fluconazole MICs were > or =16 microg/ml. Four of these patients had previously had oropharyngeal candidiasis (OPC); three had previously had episodes of cryptococcal infection, and all five treatment failure patients had high cryptococcal antigen titers in either serum or cerebrospinal fluid (titers, >1:4,000). Although 14 of the 18 patients who responded to fluconazole therapy had previously had OPC infections, they each had only a single episode of cryptococcal infection. It appears that the clinical outcome after fluconazole maintenance therapy may be better when the infecting C. neoformans strain is inhibited by lower concentrations of fluconazole for eradication (MICs, <16 microg/ml) than when the patients are infected with strains that require higher fluconazole concentrations (MICs, > or =16 microg/ml). These findings also suggest that the MICs determined by the modified NCCLS microdilution method can be potential predictors of the clinical response to fluconazole therapy and may aid in the identification of patients who will not respond to fluconazole therapy. PMID- 10817707 TI - A new class of genetic element, staphylococcus cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus. AB - We have previously shown that the methicillin-resistance gene mecA of Staphylococcus aureus strain N315 is localized within a large (52-kb) DNA cassette (designated the staphylococcal cassette chromosome mec [SCCmec]) inserted in the chromosome. By sequence determination of the entire DNA, we identified two novel genes (designated cassette chromosome recombinase genes [ccrA and ccrB]) encoding polypeptides having a partial homology to recombinases of the invertase/resolvase family. The open reading frames were found to catalyze precise excision of the SCCmec from the methicillin-resistant S. aureus chromosome and site-specific as well as orientation-specific integration of the SCCmec into the S. aureus chromosome when introduced into the cells as a recombinant multicopy plasmid. We propose that SCCmec driven by a novel set of recombinases represents a new family of staphylococcal genomic elements. PMID- 10817708 TI - OXA-24, a novel class D beta-lactamase with carbapenemase activity in an Acinetobacter baumannii clinical strain. AB - Acinetobacter baumannii RYC 52763/97, a clinical isolate involved in a prolonged nosocomial outbreak at our hospital, was resistant to all beta-lactams tested, including imipenem and meropenem, which had MICs of 128 and 256 microg/ml, respectively. This strain synthesized three beta-lactamases: a plasmid-mediated TEM-1 beta-lactamase (pI 5.4), an AmpC-type chromosomal cephalosporinase (pI 9.4), and a novel, presumptively chromosomally mediated OXA-related enzyme (pI 9.0) named OXA-24. After cloning and sequencing, the deduced amino acid sequence of the OXA-24 beta-lactamase showed 40% homology with the OXA-10 (PSE-2) and OXA 7 beta-lactamases, 39% homology with the OXA-11 and OXA-5 enzymes, and 33% homology with the LCR-1 beta-lactamase. The amino acid sequence of the OXA-24 beta-lactamase contained the STFK motif found in serine beta-lactamases, but the typical class D triad KTG was replaced by KSG and the motif YGN was replaced by FGN. The OXA-24 beta-lactamase hydrolyzed benzylpenicillin and cephaloridine but lacked activity against oxacillin, cloxacillin, and methicillin. The enzymatic activity was inhibited by chloride ions and by tazobactam (50% inhibitory concentration [IC(50)], 0.5 microM), sulbactam (IC(50), 40 microM), and clavulanic acid (IC(50), 50 microM). Carbapenem MICs for an Escherichia coli transformant (pBMB-1) expressing the cloned OXA-24 enzyme had a fourfold increase. Relative V(max)/K(m) values of 13 and 6 were obtained with imipenem and meropenem, respectively, and a positive microbiological assay result with imipenem was obtained with a purified enzymatic extract of this transformant strain. Therefore, we consider this new beta-lactamase to be involved in the carbapenem resistance of A. baumannii RYC 52763/97. PMID- 10817709 TI - Studies of the novel ketolide ABT-773: transport, binding to ribosomes, and inhibition of protein synthesis in Streptococcus pneumoniae. AB - Macrolide resistance in Streptococcus pneumoniae has been associated with two main mechanisms: target modification by Erm methyltransferases and efflux by macrolide pumps. The ketolide ABT-773, which has a 3-keto group and no L cladinose sugar, represents a new class of drugs with in vitro activity against a variety of resistant bacteria. Several approaches were undertaken to understand how ABT-773 was able to defeat resistance mechanisms. We demonstrated tighter ribosome binding of ABT-773 than erythromycin. We also showed that ABT-773 (i) accumulated in macrolide-sensitive S. pneumoniae at a higher rate than erythromycin, (ii) was able to bind with methylated ribosomes, though at lower affinities than with wild-type ribosomes, and (iii) accumulated in S. pneumoniae strains with the efflux-resistant phenotype. PMID- 10817711 TI - Relationship between capsular type, penicillin susceptibility, and virulence of human Streptococcus pneumoniae isolates in mice. AB - We examined the relationship between penicillin susceptibility, peritoneal virulence in Swiss mice, and capsular type in a selection of 122 clinical Streptococcus pneumoniae isolates belonging to 24 serotypes. Regardless of the serotype, all 32 virulent strains were susceptible to penicillin, and all 41 strains with diminished susceptibility or resistance to penicillin were avirulent. The remaining 49 strains were both susceptible to penicillin and avirulent, irrespective of the serotype. On the basis of their capsular type and pathogenic behavior, strains fell into one of four groups. In the group consisting of serotypes 1, 3, and 4 (n = 16), strains were predominantly virulent (81.3%), and all were penicillin susceptible. In the serotype 6 group (n = 32), the frequency of virulence was significantly lower (34.4 versus 81.3%, P = 0.002), and strains were predominantly penicillin susceptible (71.9%). In the group composed of serotypes 9, 14, 19, and 23 (n = 50), all strains were avirulent, and 56% had decreased susceptibility (n = 12) or resistance to (n = 16) penicillin. The fourth group was heterogenous, as it pooled 24 strains of 15 different serotypes; in this group the frequency of virulence was 33.3%, and strains were predominantly penicillin susceptible (83.3%). These data point to a complex relationship between penicillin susceptibility and virulence in mice but do not entirely separate these characteristics from the role of the capsular type. The possibility that the mechanisms conferring penicillin resistance are related to those leading to a loss of virulence is supported by these findings. PMID- 10817710 TI - Antibiotic resistance in the ECOR collection: integrons and identification of a novel aad gene. AB - The 72 Escherichia coli strains of the ECOR collection were examined for resistance to 10 different antimicrobial agents including ampicillin, tetracycline, mercury, trimethoprim, and sulfonamides. Eighteen strains were resistant to at least one of the antibiotics tested, and nearly 20% (14 of 72) were resistant to two or more. Several of the resistance determinants were shown to be carried on conjugative elements. The collection was screened for the presence of the three classes of integrons and for the sul1 gene, which is generally associated with class 1 integrons. The four strains found to carry a class 1 integron also had Tn21-encoded mercury resistance. One of the integrons encoded a novel streptomycin resistance gene, aadA7, with an attC site (or 59 base element) nearly identical to the attC site associated with the qacF gene cassette found in In40 (M.-C. Ploy, P. Courvalin, and T. Lambert, Antimicrob. Agents Chemother. 42:2557-2563, 1998). The conservation of associated attC sites among unrelated resistance cassettes is similar to arrangements found in the Vibrio cholerae superintegrons (D. Mazel, B. Dychinco, V. A. Webb, and J. Davies, Science 280:605-608, 1998) and supports the hypothesis that resistance cassettes are picked up from superintegron pools and independently assembled from unrelated genes and related attC sites. PMID- 10817712 TI - Experimental induction of fluconazole resistance in Candida tropicalis ATCC 750. AB - Candida tropicalis is less commonly isolated from clinical specimens than Candida albicans. Unlike C. albicans, which can be occasionally found as a commensal, C. tropicalis is almost always associated with the development of fungal infections. In addition, C. tropicalis has been reported to be resistant to fluconazole (FLC). To analyze the development of FLC resistance in C. tropicalis, an FLC susceptible strain (ATCC 750) (MIC = 1.0 microg/ml) was cultured in liquid medium containing increasing FLC concentrations from 8.0 to 128 microg/ml. The strain developed variable degrees of FLC resistance which paralleled the concentrations of FLC used in the medium. The highest MICs of FLC were 16, 256, and 512 microg/ml for strains grown in medium with 8.0, 32, and 128 microg of FLC per ml, respectively. Development of resistance was rapid and could be observed already after a single subculture in azole-containing medium. The resistant strains were cross-resistant to itraconazole (MIC > 1.0 microg/ml) and terbinafine (MIC > 512 microg/ml) but not to amphotericin B. Isolates grown in FLC at concentrations of 8.0 and 32 microg/ml reverted to low MICs (1.0 microg/ml) after 12 and 11 passages in FLC-free medium, respectively. The MIC for one isolate grown in FLC (128 microg/ml) (128 R) reverted to 16 microg/ml but remained stable over 60 passages in FLC-free medium. Azole-resistant isolates revealed upregulation of two different multidrug efflux transporter genes: the major facilitators gene MDR1 and the ATP-binding cassette transporter CDR1. The development of FLC resistance in vitro correlated well with the results obtained in an experimental model of disseminated candidiasis. While FLC given at 10 mg/kg of body weight/day was effective in reducing the fungal burden of mice infected with the parent strain, the same dosing regimen was ineffective in mice infected with strain 128 R. Finally, the acquisition of in vitro FLC resistance in strain 128 R was related to a loss of virulence. The results of our study elucidate important characteristics and potential mechanisms of FLC resistance in C. tropicalis. PMID- 10817713 TI - Does long-term itraconazole prophylaxis result in in vitro azole resistance in mucosal Candida albicans isolates from persons with advanced human immunodeficiency virus infection? The National Institute of Allergy and Infectious Diseases Mycoses study group. AB - The effects of prolonged itraconazole exposure on the susceptibility of Candida albicans isolates to itraconazole and fluconazole have not been well characterized. A recent placebo-controlled study of long-term itraconazole antifungal prophylaxis in persons with advanced human immunodeficiency virus infection afforded the opportunity to address this question. Mucosal Candida sp. isolates were obtained from subjects who developed oropharyngeal or esophageal candidiasis, and in vitro susceptibilities of the last isolate obtained at removal from the study as a prophylaxis failure were compared in itraconazole and placebo recipients. More subjects in the placebo group (74 of 146 [51%]) than in the itraconazole group (51 of 149 [34%]) developed mucosal candidiasis (P = 0.004). A total of 112 isolates were recovered from 56 of the 74 (76%) subjects with mucosal candidiasis assigned to the placebo group, compared to 97 isolates from 45 of the 51 (88%) subjects in the itraconazole group. C. albicans accounted for 98% of isolates in the placebo group and 89% of isolates in the itraconazole group. The itraconazole MIC at which 50% of the isolates tested were inhibited (MIC(50)) for last-episode isolates from the itraconazole group was 0.125 microg/ml compared to 0.015 microg/ml for the placebo group subjects, P = 0.0001. The MIC(50) of fluconazole for the last isolates from the itraconazole group was 1.5 microg/ml compared to 0.5 microg/ml for the placebo subjects (P = 0.005). A lower proportion of isolates recovered from subjects on itraconazole therapy were classified as susceptible to itraconazole (63%) compared to isolates from the placebo group (96%) (P = 0.001). Similarly, a lower proportion of C. albicans isolates from subjects on itraconazole therapy were susceptible to fluconazole (78%) compared to isolates from the placebo group (96%) (P = 0.01). Also, the proportion of isolates that were not fully susceptible to itraconazole or fluconazole was greater in patients assigned to the itraconazole group than the placebo group (itraconazole susceptibility, 37 and 4%, respectively (P = 0.001); fluconazole susceptibility, 23 and 4%, respectively (P = 0.01). In conclusion, long-term itraconazole prophylaxis in patients with AIDS is associated with reduction in susceptibility to itraconazole and cross-resistance to fluconazole. PMID- 10817714 TI - Recombinant green fluorescent protein-expressing human cytomegalovirus as a tool for screening antiviral agents. AB - A recombinant human cytomegalovirus (AD169-GFP) expressing green fluorescent protein was generated by homologous recombination. Infection of human fibroblast cultures with AD169-GFP virus produced stable and readily detectable amounts of GFP signals which were quantitated by automated fluorometry. Hereby, high levels of sensitivity and reproducibility could be achieved, compared to those with the conventional plaque reduction assay. Antiviral activities were determined for four reference compounds as well as a set of putative novel cytomegalovirus inhibitors. The results obtained were exactly in line with the known characteristics of reference compounds and furthermore revealed distinct antiviral activities of novel in vitro inhibitors. The fluorometric data could be confirmed by GFP-based flow cytometry and fluorescence microscopy. In addition, laboratory virus variants derived from the recombinant AD169-GFP virus provided further possibilities for study of the characteristics of drug resistance. The GFP-based antiviral assay appeared to be very reliable for measuring virus inhibitory effects in concentration- and time-dependent fashions and might also be adaptable for high-throughput screenings of cytomegalovirus-specific antiviral agents. PMID- 10817715 TI - Mild heating of amphotericin B-desoxycholate: effects on ultrastructure, in vitro activity and toxicity, and therapeutic efficacy in severe candidiasis in leukopenic mice. AB - Heated (20 min at 70 degrees C) amphotericin B-desoxycholate (hAMB-DOC) was further characterized, as was another formulation obtained after centrifugation (60 min, 3000 x g), hcAMB-DOC. Conventional AMB-DOC consisted of individual micelles (approximately 4 nm in diameter) and threadlike aggregated micelles, as revealed by cryo-transmission electron microscopy. For both hAMB-DOC and hcAMB DOC, pleiomorphic cobweb structures were observed with a mean particle size of approximately 300 nm as determined by laser diffraction. The potent antifungal activity of AMB-DOC against Candida albicans is not reduced by heating. Effective killing of C. albicans (>99.9% within 6 h) was obtained at 0.1 mg/liter with each of the AMB formulations. For AMB-DOC, hAMB-DOC, and hcAMB-DOC, cation release ((86)Rb(+)) from C. albicans of > or =50% was observed at 0.8, 0.4, and 0.4 mg/liter, respectively. After heating of AMB-DOC, toxicity was reduced 16-fold as determined by red blood cell (RBC) lysis. For AMB-DOC, hAMB-DOC, and hcAMB-DOC, hemolysis of > or =50% was observed at 6.4, 102.4, and 102.4 mg/liter, respectively. In contrast, AMB-DOC and its derivates showed similar toxicities in terms of cation release from RBC. For AMB-DOC, hAMB-DOC, and hcAMB-DOC, cation release ((86)Rb(+)) of > or =50% was observed at 1.6, 0.8, and 0.8 mg/liter, respectively. In persistently leukopenic mice with severe invasive candidiasis, higher dosages of both hAMB-DOC and hcAMB-DOC were tolerated than those of conventional AMB-DOC (3 versus 0.8 mg/kg of body weight, respectively), resulting in significantly improved therapeutic efficacy. In conclusion, this new approach of heating AMB-DOC may be of great value for further optimizing the treatment of severe fungal infections. PMID- 10817716 TI - Pharmacokinetics and tolerability of gemifloxacin (SB-265805) after administration of single oral doses to healthy volunteers. AB - Gemifloxacin (known as SB-265805 or LB-20304) is a potent, novel fluoroquinolone compound with a broad spectrum of antibacterial activity. The pharmacokinetics and tolerability of oral gemifloxacin were characterized in healthy male volunteers after a single dose of 20, 40, 80, 160, 320, 600, or 800 mg. Multiple serum and urine samples were collected and analyzed for gemifloxacin using high performance liquid chromatography with fluorescence detection. Safety assessments included vital signs, 12-lead electrocardiogram readings, hematology, clinical chemistry, urinalysis, and adverse-experience monitoring. Gemifloxacin was rapidly absorbed after all doses. Maximum concentrations of gemifloxacin in serum (C(max)) were achieved approximately 1 h after dosing, after which concentrations in serum declined in a biexponential manner. Values of C(max) and the area under the concentration-time curve in serum from 0 h to infinity (serum AUC(0 infinity)) increased linearly with dose. Serum AUC(0-infinity) values (mean +/- standard deviation) were 0.65+/-0.01, 1.28+/-0.22, 2.54+/-0.31, 5.48+/-1.24, 9.82+/-2.70, 24.4+/-7.1, and 31.4+/-7.6 microg. h/ml following 20-, 40-, 80-, 160 , 320-, 600-, and 800-mg doses, respectively. The terminal phase elimination half life was independent of dose, with an overall mean of 7.4+/-2.0 h. The profiles indicated that the pharmacokinetic profile is suitable for a once-daily dosing regimen. Approximately 25 to 40% of the administered dose was excreted unchanged in the urine, and renal clearance (ca. 150 ml/min) was independent of dose. There were no significant changes in clinical chemistry, hematology, or urinalysis parameters, vital signs, or 12-lead electrocardiogram readings in subjects, irrespective of dose. The results of these studies support the further investigation of once-daily administration of gemifloxacin. PMID- 10817717 TI - Absolute bioavailability and disposition of (-) and (+) 2'-deoxy- 3'-oxa-4' thiocytidine (dOTC) following single intravenous and oral doses of racemic dOTC in humans. AB - The purpose of this study was to characterize the pharmacokinetics and determine the absolute bioavailability of 2'-deoxy-3'-oxa-4'-thiocytidine (dOTC) (BCH 10652), a novel nucleoside analogue reverse transcriptase inhibitor, in humans. dOTC belongs to the 4'-thio heterosubstituted class of compounds and is a 1:1 mixture of its two enantiomers, (-) and (+) dOTC. Twelve healthy adult male volunteers each received oral (800-mg) and intravenous (100-mg) doses of dOTC in two study periods separated by at least 7 days. Sixteen plasma samples were obtained over 72 h and assayed for (-) and (+) dOTC, and the resultant data fit by candidate pharmacokinetic models. Data were weighted by the fitted inverse of the observation variance; model discrimination was by AIC. The pharmacokinetic model was a linear, three compartment model, with absorption occurring during one to three first-order input phases, each following a fitted lag time. The model goodness-of-fit was excellent; r(2) ranged from 0.995 to 1.0. The mean absolute bioavailabilities of (+) and (-) dOTC were 77.2% (coefficient of variation [given as a percentage] [CV%], 14) and 80.7% (CV%, 15), respectively. The median steady state volume of distribution for (+) dOTC, 74.7 (CV%, 19.2) liters/65 kg, was greater than that for (-) dOTC, 51.7 (CV%, 16.7) liters/65 kg (P<0.05). The median total clearance of (+) dOTC was less than that of (-) dOTC, 11.7 (CV%, 17.3) versus 15.4 (CV%, 18.6) liters/h/65 kg, respectively (P< 0.05). The intersubject variability of these parameters was very low. The median terminal half-life of (+) dOTC was 18.0 (CV%, 31.5) h, significantly longer than the 6.8 (CV%, 69.9) h observed for (-) dOTC (P<0.01). No serious adverse events were reported during the study. These results suggest that dOTC is well absorbed, widely distributed, and well tolerated. The terminal half-lives indicate that dosing intervals of 12 to 24 h would be reasonable. PMID- 10817719 TI - Comparison of nikkomycin Z with amphotericin B and itraconazole for treatment of histoplasmosis in a murine model. AB - Nikkomycin Z was tested both in vitro and in vivo for efficacy against Histoplasma capsulatum. Twenty clinical isolates were tested for susceptibility to nikkomycin Z in comparison to amphotericin B and itraconazole. The median MIC was 8 microg/ml with a range of 4 to 64 microg/ml for nikkomycin Z, 0.56 microg/ml with a range of 0.5 to 1.0 microg/ml for amphotericin B, and < or =0.019 microg/ml for itraconazole. Primary studies were carried out by using a clinical isolate of H. capsulatum for which the MIC of nikkomycin Z was greater than or equal to 64 microg/ml. In survival experiments, mice treated with amphotericin B at 2.0 mg/kg/dose every other day (QOD) itraconazole at 75 mg/kg/dose twice daily (BID), and nikkomycin Z at 100 mg/kg/dose BID survived to day 14, while 70% of mice receiving nikkomycin Z at 20 mg/kg/dose BID and none of the mice receiving nikkomycin Z at 5 mg/kg/dose BID survived to day 14. All vehicle control mice died by day 12. Fungal burden was assessed on survivors. Mice treated with nikkomycin Z at 20 and 100 mg/kg/dose BID had significantly higher CFUs per gram of organ weight in quantitative cultures and higher levels of Histoplasma antigen in lung and spleen homogenates than mice treated with amphotericin B at 2.0 mg/kg/dose QOD or itraconazole at 75 mg/kg/dose BID. Studies also were carried out with a clinical isolate for which the MIC of nikkomycin Z was 4 microg/ml. All mice treated with amphotericin B at 2.0 mg/kg/dose QOD; itraconazole at 75 mg/kg/dose BID; and nikkomycin Z at 100, 20, and 5 mg/kg/dose BID survived until the end of the study at day 17 postinfection, while 30% of the untreated vehicle control mice survived. Fungal burden assessed on survivors showed similar levels of Histoplasma antigen in lung and spleen homogenates of mice treated with amphotericin B at 2.0 mg/kg/dose QOD; itraconazole at 75 mg/kg/dose BID; and nikkomycin Z at 100, 20, and 5 mg/kg/dose BID. The three surviving vehicle control mice had significantly higher antigen levels in lung and spleen than other groups (P<0.05). The efficacy of nikkomycin Z at preventing mortality and reducing fungal burden correlates with in vitro susceptibility. PMID- 10817718 TI - Phenotypic expression of oxacillin resistance in Staphylococcus epidermidis: roles of mecA transcriptional regulation and resistant-subpopulation selection. AB - The MICs for many oxacillin-resistant (OR) Staphylococcus epidermidis (ORSE) strains are below the Staphylococcus aureus methicillin or oxacillin resistance breakpoint. The difficulty detecting the OR phenotype in S. epidermidis may be due to extreme heterotypy in resistance expression and/or transcriptional repression of mecA, the OR gene, by MecI. To determine the role of these factors in the phenotypic expression of ORSE, 17 geographically diverse mecI(+) ORSE isolates representing 14 distinct pulsed-field gel electrophoresis pulse types (>3 band differences) were investigated. Thirteen of the 14 types contained mecI and mecA promoter-operator sequences known to be associated with maximal mecA repression, and in all isolates, mecA transcription was repressed. All 17 were heterotypic in their resistance expression. Oxacillin MICs ranged from 1 to 128 microg/ml and increased for 16 of 17 isolates after beta-lactam induction. Allelic replacement inactivation of mecI in three isolates similarly resulted in a four- to sevenfold increase in MIC. In the two of these three isolates producing beta-lactamase, mecA transcription was regulated by both mecI and beta lactamase regulatory sequences. Heterotypic expression of resistance in these three isolates was unaffected by either beta-lactam induction or mecI inactivation. However, prolonged incubation in concentrations of oxacillin just sufficient to produce a lag in growth (0.5 to 1.0 microg/ml) converted the population resistance expression from heterotypic to homotypic. Homotypic conversion could also be demonstrated in microtiter wells during MIC determinations in one isolate for which the MIC was high. We conclude that the phenotypic expression of S. epidermidis OR in broth can be affected both by mecA transcriptional regulation and by subpopulation resistance expression. PMID- 10817720 TI - Inhibitors of sterol biosynthesis and amphotericin B reduce the viability of pneumocystis carinii f. sp. carinii. AB - Pneumocystis carinii synthesizes sterols with a double bond at C-7 of the sterol nucleus and an alkyl group with one or two carbons at C-24 of the side chain. Also, some human-derived Pneumocystis carinii f. sp. hominis strains contain lanosterol derivatives with an alkyl group at C-24. These unique sterols have not been found in other pathogens of mammalian lungs. Thus, P. carinii may have important differences in its susceptibility to drugs known to block reactions in ergosterol biosynthesis in other fungi. In the present study, inhibitors of 3 hydroxy-3-methyglutaryl coenzyme A reductase, squalene synthase, squalene epoxidase, squalene epoxide-lanosterol cyclase, lanosterol demethylase, Delta(8) to Delta(7) isomerase, and S-adenosylmethionine:sterol methyltransferase were tested for their effects on P. carinii viability as determined by quantitation of cellular ATP levels in a population of organisms. Compounds within each category varied in inhibitory effect; the most effective included drugs targeted at squalene synthase, squalene epoxide-lanosterol cyclase, and Delta(8) to Delta(7) isomerase. Some drugs that are potent against ergosterol-synthesizing fungi had little effect against P. carinii, suggesting that substrates and/or enzymes in P. carinii sterol biosynthetic reactions are distinct. Amphotericin B is ineffective in clearing P. carinii infections at clinical doses; however, this drug apparently binds to sterols and causes permeability changes in P. carinii membranes, since it reduced cellular ATP levels in a dose-dependent fashion. PMID- 10817721 TI - Determinants of ceftazidime clearance by continuous venovenous hemofiltration and continuous venovenous hemodialysis. AB - Although several dosage adjustment regimens have been proposed, there is little quantitative information to guide the initiation of ceftazidime therapy in patients who are receiving continuous renal replacement therapy. To determine the clearance of ceftazidime by continuous venovenous hemofiltration (CVVH) and continuous venovenous hemodialysis (CVVHD), we performed controlled clearance studies with stable hemodialysis patients with three hemofilters: a 0.6-m(2) acrylonitrile copolymer (AN69; Hospal) filter, a 2.1-m(2) polymethylmethacrylate filter (PMMA; Toray) filter and a 0.65-m(2) polysulfone (PS; Fresenius) filter. Subjects received 1,000 mg of ceftazidime intravenously prior to the start of a clearance study. The concentration of ceftazidime in multiple plasma and dialysate or ultrafiltrate samples was determined by high-performance liquid chromatography. The diffusional clearances (CI(diffusion)) and sieving coefficients of ceftazidime were compared by a mixed-model repeated-measures analysis of variance with filter and blood, dialysate inflow, or ultrafiltration rate as the main effect and the patient as a random effect. The fraction of ceftazidime bound to plasma proteins was 17%+/-7% (range, 10 to 25%). The clearances of ceftazidime, urea, and creatinine by CVVHD were essentially constant at blood flow rates of 75 to 250 ml/min for all three filters. Significant linear relationships (P<0.0001) were observed between CI(diffusion) of ceftazidime and clearance of urea for all three filters: AN69 (slope = 0.83), PMMA (slope = 0.89), and PS (slope = 1.03). Ceftazidime clearance was membrane independent during CVVH and CVVHD. CVVH and CVVHD can significantly augment the clearance of ceftazidime. Dosing strategies for initiation of ceftazidime therapy in patients receiving CVVH and CVVHD are proposed. PMID- 10817722 TI - In vivo antimalarial activity of the beta-carboline alkaloid manzamine A. AB - Manzamine A, a beta-carboline alkaloid present in several marine sponge species, inhibits the growth of the rodent malaria parasite Plasmodium berghei in vivo. More than 90% of the asexual erythrocytic stages of P. berghei were inhibited after a single intraperitoneal injection of manzamine A into infected mice. A remarkable aspect of manzamine A treatment is its ability to prolong the survival of highly parasitemic mice, with 40% recovery 60 days after a single injection. Oral administration of an oil suspension of manzamine A also produced significant reductions in parasitemia. The plasma manzamine A concentration peaked 4 h after injection and remained high even at 48 h. Morphological changes of P. berghei were observed 1 h after treatment of infected mice. (-)-8-Hydroxymanzamine A also displayed antimalarial activity, whereas manzamine F, a ketone analog of manzamine A, did not. Our results suggest that manzamine A and (-)-8 hydroxymanzamine A are promising new antimalarial agents. PMID- 10817723 TI - Increasing bacterial resistance in pediatric acute conjunctivitis (1997-1998). AB - We sought to determine the current level of resistance in Haemophilus influenzae and Streptococcus pneumoniae, the primary pathogens of pediatric conjunctivitis. Between January 1997 and March 1998, we prospectively cultured acute conjunctivitis in 250 ambulatory pediatric patients from rural Kentucky whose average age was 24.3 months. In those 250 cases, 106 H. influenzae (42% of the total) and 75 S. pneumoniae (30% of the total) pathogens were isolated, with no growth or no pathogen resulting in 79 cases (32% of the total). Beta-lactamase was detected in 60 (69%) of 87 tested strains of H. influenzae. Among 65 isolates of S. pneumoniae tested for penicillin susceptibility, 44 (68%) were susceptible, 17 (26%) were resistant, and 4 (6%) were intermediate. Conjunctivitis with acute otitis media was observed in 97 patients (39%), and H. influenzae was recovered in 57% of these 97 cases. As for in vitro activity, ciprofloxacin, ofloxacin, and tetracycline were the most active; and gentamicin, tobramycin, polymyxin B trimethoprim, and polymyxin B-neomycin were intermediately active. Sulfamethoxazole possessed no activity against either pathogen. Beta-lactamase production was detected in 69% of H. influenzae strains, which still remains the primary causative pathogen of both conjunctivitis and conjunctivitis-otitis syndrome. Penicillin-nonsusceptible S. pneumoniae was observed in 32% of 65 patients with S. pneumoniae conjunctivitis, with most strains being penicillin resistant. PMID- 10817724 TI - Use of drug effect interaction modeling with Monte Carlo simulation to examine the impact of dosing interval on the projected antiviral activity of the combination of abacavir and amprenavir. AB - The delineation of optimal regimens for combinations of agents is a difficult problem, in part because, to address it, one needs to (i) have effect relationships between the pathogen in question and the drugs in the combination, (ii) have knowledge of how the drugs interact (synergy, antagonism, and additivity), and (iii) address the issue of true between-patient variability in pharmacokinetics for the drugs in the population. We have developed an approach which employs a fully parametric assessment of drug interaction using the equation of W. R. Greco, G. Bravo, and J. C. Parsons (Pharmacol. Rev. 47:331-385, 1995) to generate an estimate of effects for the two drugs and have linked this approach to a population simulator, using Monte Carlo methods, which produce concentration-time profiles for the drugs in combination. This software automatically integrates the effect over a steady-state dosing interval and produces an estimate of the mean effect over a steady-state interval for each simulated subject. In this way, doses and schedules can be easily evaluated. This software allows for a rational choice of dose and schedule for evaluation in clinical trials. We evaluated different schedules of administration for the combination of the nucleoside analogue abacavir plus the human immunodeficiency virus type 1 protease inhibitor amprenavir. Amprenavir was simulated as either 800 mg every 8 h (q8h) or 1,200 mg q12h, each along with 300 mg q12h of abacavir. Both regimens produced excellent effects over the simulated population of 500 subjects, with average percentages of maximal effect (as determined from the in vitro assays) of 90.9%+/- 11.4% and 80.9%+/-18.6%, respectively. This difference is statistically significant (P<<0.001). In addition, 68.8 and 46.0% of the population had an average percentage of maximal effect which was greater than or equal to 90% for the two regimens. We can conclude that the combination of abacavir plus amprenavir is a potent combination when it is given on either schedule. However, the more fractionated schedule for the protease inhibitor produced significantly better effects in combination. Clinicians need to explicitly balance the improvement in antiviral effect seen with the more fractionated regimen against the loss of compliance attendant to the use of such a regimen. This approach may be helpful in the preclinical evaluation of multidrug anti-infective regimens. PMID- 10817725 TI - vanC cluster of vancomycin-resistant Enterococcus gallinarum BM4174. AB - Glycopeptide-resistant enterococci of the VanC type synthesize UDP-muramyl pentapeptide[D-Ser] for cell wall assembly and prevent synthesis of peptidoglycan precursors ending in D-Ala. The vanC cluster of Enterococcus gallinarum BM4174 consists of five genes: vanC-1, vanXY(C), vanT, vanR(C), and vanS(C). Three genes are sufficient for resistance: vanC-1 encodes a ligase that synthesizes the dipeptide D-Ala-D-Ser for addition to UDP-MurNAc-tripeptide, vanXY(C) encodes a D,D-dipeptidase-carboxypeptidase that hydrolyzes D-Ala-D-Ala and removes D-Ala from UDP-MurNAc-pentapeptide[D-Ala], and vanT encodes a membrane-bound serine racemase that provides D-Ser for the synthetic pathway. The three genes are clustered: the start codons of vanXY(C) and vanT overlap the termination codons of vanC-1 and vanXY(C), respectively. Two genes which encode proteins with homology to the VanS-VanR two-component regulatory system were present downstream from the resistance genes. The predicted amino acid sequence of VanR(C) exhibited 50% identity to VanR and 33% identity to VanR(B). VanS(C) had 40% identity to VanS over a region of 308 amino acids and 24% identity to VanS(B) over a region of 285 amino acids. All residues with important functions in response regulators and histidine kinases were conserved in VanR(C) and VanS(C), respectively. Induction experiments based on the determination of D,D-carboxypeptidase activity in cytoplasmic extracts confirmed that the genes were expressed constitutively. Using a promoter-probing vector, regions upstream from the resistance and regulatory genes were identified that have promoter activity. PMID- 10817726 TI - Pharmacokinetics and safety of AMD-3100, a novel antagonist of the CXCR-4 chemokine receptor, in human volunteers. AB - AMD-3100, a bicyclam, is a novel agent that uniquely inhibits the entry of human immunodeficiency virus type 1 (HIV-1) into CD4(+) T cells via selective blockade of the chemokine CXCR-4 receptor. Twelve healthy volunteers were given AMD-3100 as a single 15-min intravenous infusion at 10, 20, 40, or 80 microg/kg. Five subjects also received a single subcutaneous injection of AMD-3100 (40 or 80 microg/kg). Three subjects received two escalating oral doses each (80 and 160 microg/kg). All subjects tolerated their dose(s) well without any grade 2 toxicity or dose adjustment. Six subjects experienced mild, transient symptoms, primarily gastrointestinal in nature and not dose related. All subjects experienced a dose-related elevation of the white blood cell count, from 1.5 to 3.1 times the baseline, which returned to the baseline 24 h after dosing. AMD 3100 demonstrated dose proportionality for the maximum drug concentration in serum (C(max)) and the area under the concentration-time curve from 0 h to infinity (AUC(0-infinity)) over the entire dose range. At the highest intravenous dose (80 microg/kg), the median C(max) was 515 (range, 470 to 521) ng/ml and the AUC(0-infinity) was 1,044 (range, 980 to 1,403) ng-h/ml. The median systemic absorption after subcutaneous dosing was 87% (range, 67 to 106%). No drug was detectable in the blood following oral dosing. Using a two-compartment model, the median pharmacokinetic parameter estimates (ranges) were as follows: volume of distribution, 0.34 (0. 27 to 0.36) liter/kg; clearance, 1.30 (0.97 to 1.34) liters/h; elimination half-life, 3.6 (3.5 to 4.9) h. After a single, well tolerated intravenous dose of AMD-3100, concentrations were sustained for 12 h above the in vitro antiretroviral 90% inhibitory concentrations and for 8 h above antiviral concentrations identified in the SCID-hu Thy/Liv mouse model of HIV infection. PMID- 10817727 TI - Pharmacokinetics of ciprofloxacin in the human eye: a clinical study and population pharmacokinetic analysis. AB - Ciprofloxacin, a fluoroquinolone antibiotic active against a wide variety of bacteria, is one of a few antibiotics which enters the human eye after oral administration. However, little is known about its pharmacokinetics in the human eye. One or two oral doses of 750 mg of ciprofloxacin (at a 12-h interval) were administered to 48 patients at various times prior to ocular surgery. Clotted blood, aqueous, and vitreous were collected at surgery, and the concentrations of ciprofloxacin were assayed by high-performance liquid chromatography. Our data were combined with those of others, and a population pharmacokinetic analysis was conducted. The concentrations of ciprofloxacin in both aqueous and vitreous were lower than those in serum and peaked at a later time. The pharmacokinetics of ciprofloxacin in aqueous and vitreous were fitted to a compartmental model in which the antibiotic was transferred into and out of the two compartments (aqueous and vitreous) by first-order processes. Population pharmacokinetic software, P-Pharm, was used to calculate the mean half-lives of the loss of ciprofloxacin from aqueous and vitreous, which were 3.5 and 5.3 h, respectively. At steady state, the mean ratios of then concentrations in aqueous and vitreous to the concentrations in serum were 23 and 17%, respectively. After the administration of one or two doses of 750 mg of ciprofloxacin, the concentrations in both aqueous and vitreous in a number of patients were lower than the MICs at which 90% of isolates are inhibited (0.5 mg/liter) for common intraocular bacterial pathogens. Simulations of concentrations in the eye after the administration of higher doses (1,500 mg of ciprofloxacin as a single dose, two doses of 750 mg 2 h apart, and 750 mg every 6 h) indicated that in approximately 20% of patients the concentrations would still be below 0.5 mg/liter. Although oral ciprofloxacin may be a beneficial adjunctive therapy, the use of oral ciprofloxacin alone may not be adequate for perioperative prophylaxis or for treatment of bacterial endophthalmitis. PMID- 10817728 TI - Therapeutic responses to different antimalarial drugs in vivax malaria. AB - The therapeutic responses to the eight most widely used antimalarial drugs were assessed in 207 adult patients with Plasmodium vivax malaria. This parasite does not cause marked sequestration, so parasite clearance can be used as a direct measure of antimalarial activity. The activities of these drugs in descending order were artesunate, artemether, chloroquine, mefloquine, quinine, halofantrine, primaquine, and pyrimethamine-sulfadoxine (PS). Therapeutic responses to PS were poor; parasitemias did not clear in 5 of the 12 PS-treated patients, whereas all the other patients made an initial recovery. Of 166 patients monitored for > or =28 days, 35% had reappearance of vivax malaria 11 to 65 days later and 7% developed falciparum malaria 5 to 21 days after the start of treatment. There were no significant differences in the times taken for vivax malaria reappearance among the different groups except for those given mefloquine and chloroquine, in which all vivax malaria reappearances developed >28 days after treatment, suggesting suppression of the first relapse by these slowly eliminated drugs. There was no evidence of chloroquine resistance. The antimalarial drugs vary considerably in their intrinsic activities and stage specificities of action. PMID- 10817729 TI - Pharmacokinetic interaction of abacavir (1592U89) and ethanol in human immunodeficiency virus-infected adults. AB - While in vitro results at clinically relevant concentrations do not predict abacavir (1592U89) interactions with drugs highly metabolized by cytochrome P450, the potential does exist for a pharmacokinetic interaction between abacavir and ethanol, as both are metabolized by alcohol dehydrogenase. Twenty-five subjects were enrolled in an open-label, randomized, three-way-crossover, phase I study of human immunodeficiency virus-infected male subjects. The three treatments were administration of (i) 600 mg of abacavir, (ii) 0.7 g of ethanol per kg of body weight, and (iii) 600 mg of abacavir and 0.7 g of ethanol per kg. Twenty-four subjects completed the study with no unexpected adverse events reported. Ethanol pharmacokinetic parameters were unchanged with abacavir coadministration. The geometric least squares mean area under the concentration curve extrapolated to infinite time for abacavir increased 41% (from 11.07 to 15.62 microg. h/ml), and the half-life increased 26% (from 1.42 to 1.79 h) in the presence of ethanol (mean ethanol maximum concentration in plasma of 498 microg/ml). The percentages of abacavir dose recovered in urine as abacavir and its two major metabolites were each altered in the presence of ethanol, but there was no change in the total percentage ( approximately 50%) of administered dose recovered in the 12-h collection interval. In conclusion, while a single 600-mg dose of abacavir does not alter blood ethanol concentration, ethanol does increase plasma abacavir concentrations. PMID- 10817730 TI - Disposition of intravenous pyrimethamine in healthy volunteers. AB - A proportion of patients with AIDS and toxoplasmic encephalitis (TE) sustain low plasma pyrimethamine concentrations during oral treatment, possibly because of incomplete and variable bioavailability. We wanted to develop a safe, practicable intravenous (i.v.) formulation of pyrimethamine and characterize its disposition in healthy volunteers. A neutral, aqueous, sterile solution of pyrimethamine was produced and presented in sealed glass ampoules. Pyrimethamine (1 mg/kg) was given to eight healthy male volunteers by i.v. infusion over 2 h, and blood was sampled over a 2 week period. Pyrimethamine levels in plasma were measured by high-performance liquid chromatography. The drug was well tolerated by all volunteers, and there were no changes in vital signs, electrocardiogram, hematology, or biochemical parameters. The maximum pyrimethamine concentration of 2,089+/-565 ng ml(-1) (mean +/- standard deviation) was achieved shortly after the end of the infusion; thereafter, concentrations declined in a log-linear manner, with a half-life of 140+/-31 h. PMID- 10817731 TI - In vitro susceptibility tests for cationic peptides: comparison of broth microdilution methods for bacteria that grow aerobically. AB - The in vitro susceptibilities of 90 clinical isolates of gram-positive and gram negative aerobic bacteria to six cationic peptides, buforin II, cecropin P1, indolicidin, magainin II, nisin, and ranalexin, were evaluated by two broth microdilution methods. The first method was performed according to the procedures outlined by the National Committee for Clinical Laboratory Standards for bacteria that grow aerobically, while the second was performed according to the procedures recently proposed by the R. E. W. Hancock laboratory for testing antimicrobial peptides. Overall, the first method produced MICs two- and fourfold higher than the second method. PMID- 10817732 TI - Interactions of ofloxacin and erythromycin with the multidrug resistance protein (MRP) in MRP-overexpressing human leukemia cells. AB - To investigate interactions between the multidrug resistance protein (MRP) and antimicrobial agents, we examined the effects of 12 agents on vincristine sensitivity and efflux of the calcein acetoxy-methyl ester (calcein-AM) of a MRP substrate in MRP-overexpressing cells. Only ofloxacin and erythromycin enhanced sensitivity with increased intracellular vincristine accumulation and inhibited the calcein-AM efflux. Our findings suggest that the two agents are possible MRP substrates and may competitively inhibit MRP function as a drug efflux pump. PMID- 10817734 TI - Antibiotic susceptibility patterns of Neisseria meningitidis isolates from patients and asymptomatic carriers. AB - The activities of seven antimicrobial agents used for treatment and prophylaxis of meningococcal disease was investigated against 901 Neisseria meningitidis isolates, 112 of which were recovered from patients and 789 of which were recovered from asymptomatic carriers. The proportions of isolates with decreased susceptibility to penicillin were 55.3 and 39.0%, respectively. Penicillin- and ampicillin-intermediate strains were more common among serogroup C meningococci than among non-serogroup C meningococci from both patients and carriers. PMID- 10817733 TI - Characterization of a Mycobacterium smegmatis mutant that is simultaneously resistant to D-cycloserine and vancomycin. AB - A mutant of Mycobacterium smegmatis has been isolated that is simultaneously resistant to both D-cycloserine (D-CS) and vancomycin. Genetic complementation with a PBP4 homolog restores sensitivity to both drugs. Resistance to D-CS and vancomycin in this mutant is most likely due to a novel mechanism involving peptidoglycan assembly at the cell surface. PMID- 10817735 TI - Ineffectiveness of echinacea for prevention of experimental rhinovirus colds. AB - The purpose of this study was to assess the effectiveness of echinacea for the prevention of experimental rhinovirus colds. Infection occurred in 44 and 57% and illness occurred in 36 and 43% of the echinacea- and placebo-treated subjects, respectively. This preparation of echinacea had no significant effect on either the occurrence of infection or the severity of illness. PMID- 10817736 TI - Comparative in vitro activities of clinafloxacin and trovafloxacin against 1,000 isolates of bacteroides fragilis group: effect of the medium on test results. AB - The in vitro antibacterial activities of clinafloxacin, trovafloxacin, ciprofloxacin, and cefoxitin against 1,000 clinical isolates of Bacteroides fragilis group were compared by agar dilution in brucella blood agar (BBA) and Wilkins Chalgren agar (WCA). Significantly higher geometric mean MICs for the three quinolones and cefoxitin (P<0.001) were obtained in BBA than in WCA. Regardless of medium, clinafloxacin was slightly more active than trovafloxacin. The activity of clinafloxacin and trovafloxacin was greater than that of cefoxitin against B. distasonis, B. ovatus, and B. thetaiotaomicron but lower against B. vulgatus. High cross resistance between trovafloxacin and clinafloxacin was observed. PMID- 10817737 TI - Mutations at four distinct regions of the rpoB gene can reduce the susceptibility of Helicobacter pylori to rifamycins. AB - The transformation of a panel of rpoB mutations (codons 525 to 586) from rifampin resistant donor strains into Helicobacter pylori 2802A confirmed the MICs associated with the respective mutations. RpoB V149X random mutations were generated and induced different levels of resistance, depending on the replacement amino acid. Mutagenesis of the rpoB gene at codon position 701 (R701H) induced low-level resistance. PMID- 10817739 TI - Activities of taurolidine in vitro and in experimental enterococcal endocarditis. AB - In vitro, the antimicrobial agent taurolidine inhibited virtually all of the bacteria tested, including vancomycin-resistant enterococci, oxacillin-resistant staphylococci, and Stenotrophomonas maltophilia, at concentrations between 250 and 2,000 microg/ml. Taurolidine was not effective in experimental endocarditis. While it appears unlikely that this antimicrobial would be useful for systemic therapy, its bactericidal activity and the resistance rates found (<10(-9)) are favorable indicators for its possible development for topical use. PMID- 10817738 TI - In vitro activities of membrane-active peptides alone and in combination with clinically used antimicrobial agents against Stenotrophomonas maltophilia. AB - The in vitro activities of buforin II, cecropin P1, and magainin II, alone and in combination with six clinically used antimicrobial agents, against 12 clinical isolates of Stenotrophomonas maltophilia were investigated. Antimicrobial activities were measured by MIC and time-kill studies. The isolates were susceptible to the peptides at concentrations in the range of 0.50 to 16 microg/ml. Synergy was observed when the peptides were combined with polymyxin E, meropenem, ceftazidime, piperacillin, and clarithromycin. PMID- 10817740 TI - A new SHV-derived extended-spectrum beta-lactamase (SHV-24) that hydrolyzes ceftazidime through a single-amino-acid substitution (D179G) in the -loop. AB - A new SHV-derived extended-spectrum beta-lactamase (SHV-24) conferring high-level resistance to ceftazidime but not cefotaxime and cefazolin was identified in Japan. This enzyme was encoded by a transferable 150-kb plasmid from an Escherichia coli clinical isolate. The pI and K(m) for CAZ of this enzyme were 7.5 and 30 microM, respectively. SHV-24 was found to have a D179G substitution in the Omega-loop of the enzyme. PMID- 10817741 TI - Efficacy of FK463, a (1,3)-beta-D-glucan synthase inhibitor, in disseminated azole-resistant candida albicans infection in mice. AB - The efficacy of FK463, a new (1,3)-beta-D-glucan synthase inhibitor, against azole-resistant Candida albicans strains has been studied. The MIC of FK463 was lower than those of azoles and amphotericin B against CDR1-expressing C26 and CaMDR-expressing C40 strains. All mice treated with FK463 (1 mg/kg) survived disseminated murine candidiasis. The fungal burden in the kidney after 6 days was markedly reduced after therapy with FK463 and amphotericin B sodium deoxycholate, and plasma (1,3)-beta-D-glucan concentration was found to be lower in FK463 treated mice. In our study, FK463 was found to be a potent antifungal agent against disseminated infection with azole-resistant C. albicans. PMID- 10817742 TI - Quinolone resistance in enterotoxigenic Escherichia coli causing diarrhea in travelers to India in comparison with other geographical areas. AB - Enterotoxigenic Escherichia coli isolates were identified as a cause of traveler's diarrhea in 82 of 520 (16%) patients and tested for resistance to seven antimicrobial agents. Thirty patients (36%) needed antimicrobial therapy: 17 (56%) for persistence of symptoms and 13 (44%) for severity of symptoms. Ampicillin, tetracycline, and trimethoprim-sulfamethoxazole resistance was high. Chloramphenicol showed moderate activity, and amoxicillin plus clavulanic acid, nalidixic acid, and ciprofloxacin showed very good activity. Five nalidixic acid resistant strains were isolated, four from patients visiting India. PMID- 10817744 TI - Failure of azithromycin in treatment of Brill-Zinsser disease. AB - Two patients suffering from Brill-Zinsser disease were treated with azithromycin, which did not prove effective. Rickettsia prowazekii, the agent causing Brill Zinsser disease, cannot be treated with azithromycin. Both patients had epidemiological features consistent with and a clinical course typical of the disease. The diagnosis of Brill-Zinsser disease was serologically confirmed. PMID- 10817743 TI - In vitro activities of voriconazole, itraconazole, and amphotericin B against Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum. AB - The in vitro activity of voriconazole was compared to those of itraconazole and amphotericin B against the mold forms of 304 isolates of three dimorphic fungi, Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum. MICs were determined by a broth microdilution adaptation of the National Committee for Clinical Laboratory Standards M27-A procedure. RPMI 1640 medium was used for tests with voriconazole and itraconazole, whereas Antibiotic Medium 3 with 2% glucose was used for amphotericin B. Minimum fungicidal concentrations (MFCs) were also determined. Amphotericin B was active against all three dimorphic fungi, with MICs at which 90% of the isolates tested are inhibited (MIC(90)s) of 0.5 to 1 microg/ml. Itraconazole had MIC(90)s of 0.06 microg/ml for H. capsulatum, 0.125 microg/ml for B. dermatitidis, and 1 microg/ml for C. immitis. The MIC(90)s of voriconazole were 0.25 microg/ml for all three fungi. Amphotericin B was fungicidal for B. dermatitidis and H. capsulatum with MFCs at which 90% of strains tested are killed (MFC(90)s) of 0.5 and 2 microg/ml, respectively. It was less active against C. immitis, with MFCs ranging from 0.5 to >16 microg/ml. Voriconazole and itraconazole were lethal for most isolates of B. dermatitidis, with MFC(50)s and MFC(90)s of 0.125 and 4 microg/ml, respectively. Both azoles were fungicidal for some isolates of H. capsulatum, with MFC(50)s of 2 and 8 microg/ml for itraconazole and voriconazole, respectively; neither had a lethal effect upon C. immitis. Our results suggest that voriconazole possesses promising activity against these important human pathogens. PMID- 10817745 TI - Antileishmanial activities of stearylamine-bearing liposomes. AB - Here we report the activity of liposomes comprising egg phosphatidylcholine (PC) and stearylamine (SA) against Leishmania donovani parasites. Both promastigotes and intracellular amastigotes in vitro and in vivo were susceptible to SA-PC liposomes. A single dose of 55 mg of SA-PC liposomes/animal could significantly reduce the hepatic parasite burden by 85 and 68% against recent and established experimental visceral leishmaniasis, respectively, suggesting their strong therapeutic potential. PMID- 10817746 TI - Cyclic AMP and fluconazole resistance in Saccharomyces cerevisiae. AB - Cyclic AMP (cAMP) is involved in the stress response in Saccharomyces cerevisiae. We show that cAMP is required for resistance to fluconazole in S. cerevisiae. In addition, activation of Ras2, a regulator of cAMP generation, results in some protection from fluconazole toxicity in a fashion independent of the efflux transporter Pdr5p. PMID- 10817748 TI - Postantibiotic suppression of growth of erythromycin A-susceptible and -resistant gram-positive bacteria by the ketolides telithromycin (HMR 3647) and HMR 3004. AB - We investigated the in vitro postantibiotic effects (PAEs) of the ketolides telithromycin (HMR 3647) and HMR 3004 and analyzed the results using the sigmoid E(max) model. Mean maximum telithromycin PAEs against erythromycin A-susceptible strains of Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae were 3. 7, 8.9, and 9.7 h, respectively, while maximum PAEs for erythromycin A-resistant strains were much shorter. Mean maximum HMR 3004 PAEs were 3.2 to 4.4 h for all species. PMID- 10817747 TI - Identification and characterization of the penicillin-binding protein 2a of Streptococcus pneumoniae and its possible role in resistance to beta-lactam antibiotics. AB - To further understand the role of penicillin-binding protein 2a (PBP 2a) of Streptococcus pneumoniae in penicillin resistance, we confirmed the identity of the protein as PBP 2a. The PBP 2a protein migrated electrophoretically to a position corresponding to that of PBP 2x, PBP 2a, and PBP 2b of S. pneumoniae and was absent in a pbp2a insertional mutant of S. pneumoniae. We found that the affinities of PBP 2a for penicillins were lower than for cephalosporins and a carbapenem. When compared with other S. pneumoniae PBPs, PBP 2a exhibited lower affinities for beta-lactam antibiotics, especially penicillins. Therefore, PBP 2a is a low-affinity PBP for beta-lactam antibiotics in S. pneumoniae. PMID- 10817749 TI - Single-oral-dose azithromycin prophylaxis against experimental streptococcal or staphylococcal aortic valve endocarditis. AB - Azithromycin and ampicillin protected 94 and 72% of animals challenged with Streptococcus oralis, respectively (P = 0.177), while azithromycin and vancomycin protected 59 and 94% of the methicillin-resistant Staphylococcus aureus (MRSA) challenged animals, respectively (P = 0.018). Azithromycin is effective in preventing experimental streptococcal endocarditis, but against MRSA it is less effective than vancomycin. PMID- 10817750 TI - Effect of oral administration of emtricitabine on woodchuck hepatitis virus replication in chronically infected woodchucks. AB - Emtricitabine [(-)FTC] [(-)-beta-2', 3'-dideoxy-5-fluoro-3'-thiacytidine] has been shown to be an effective inhibitor of hepatitis B virus (HBV) in cell culture, with a potency and selectivity that are essentially identical to those of lamivudine. The antiviral activity of oral administration of (-)FTC against WHV replication in chronically infected woodchucks, an established and predictive model for antiviral therapy against HBV, was examined in a placebo-controlled study. (-)FTC significantly reduced viremia and intrahepatic WHV replication in a dose-dependent manner that was comparable to the antiviral activity of lamivudine observed in previous studies conducted by our laboratories. No effect on the levels of hepatic WHV RNA or the levels of woodchuck hepatitis surface antigen or anti-woodchuck hepatitis surface and core antibodies in the serum of the treated animals was observed. No evidence of drug-related toxicity was observed in any of the animals treated. PMID- 10817751 TI - Effect of azithromycin plus rifampin versus amoxicillin alone on eradication and inflammation in the chronic course of Chlamydia pneumoniae pneumonitis in mice. AB - The effects of treatment with azithromycin plus rifampin (A+R), amoxicillin (A), or placebo (P) on the chronic course of experimental Chlamydia pneumoniae pneumonitis in mice were assessed by culture, PCR, and immunocytochemistry as well as by degree of inflammation in lung tissue. Eradication of the pathogen was significantly more frequent and inflammation in tissue was significantly reduced after treatment with A+R compared to after treatment with A or P. Combination therapy with azithromycin plus rifampin showed favorable effects in the chronic course of C. pneumoniae pneumonitis. PMID- 10817752 TI - Cadherin superfamily genes: functions, genomic organization, and neurologic diversity. PMID- 10817753 TI - Wnt signaling maintains the hair-inducing activity of the dermal papilla. AB - The formation of the hair follicle and its cyclical growth, quiescence, and regeneration depend on reciprocal signaling between its epidermal and dermal components. The dermal organizing center, the dermal papilla (DP), regulates development of the epidermal follicle and is dependent on signals from the epidermis for its development and maintenance. GFP specifically expressed in DP cells of a transgenic mouse was used to purify this population and study the signals required to maintain it. We demonstrate that specific Wnts, but not Sonic hedgehog (Shh), maintain anagen-phase gene expression in vitro and hair inductive activity in a skin reconstitution assay. PMID- 10817754 TI - H19 and Igf2 monoallelic expression is regulated in two distinct ways by a shared cis acting regulatory region upstream of H19. AB - H19 and Igf2 are expressed in a monoallelic fashion from the maternal and paternal chromosomes, respectively. A region upstream of H19 has been shown to regulate such imprinted expression of both genes in cis. We have taken advantage of a loxP/cre recombinase-based strategy to delete this region in mice in a conditional manner to determine the temporal requirement of the upstream region in initiating and maintaining the imprinted expression of H19 and Igf2. Analysis of allele-specific expression of H19 and Igf2 and DNA methylation at the H19 promoter demonstrates that this region controls the monoallelic expression of the two genes in different ways, suggesting that it harbors two functionally distinct regulatory elements. Continued presence of the region is required to silence maternal Igf2 in accordance with its proposed role as an insulator. However, it does not have a direct role in keeping the paternal H19 promoter silenced. Instead, on the paternal chromosome, the upstream element mediates epigenetic modifications of the H19 promoter region during development, leading to transcriptional silencing of H19. Thereafter, its presence is redundant for preventing transcription. Presently, this temporal requirement of the silencing element appears to be a unique cis activity in the mammalian system. However, it is likely that other cis-acting elements, positive and negative, have the ability to effect stable changes in the chromatin structure and are not constantly required to give signals to the transcriptional machinery. PMID- 10817755 TI - The something about silencing protein, Sas3, is the catalytic subunit of NuA3, a yTAF(II)30-containing HAT complex that interacts with the Spt16 subunit of the yeast CP (Cdc68/Pob3)-FACT complex. AB - We have purified and characterized a Gcn5-independent nucleosomal histone H3 HAT complex, NuA3 (Nucleosomal Acetyltransferase of histone H3). Peptide sequencing of proteins from the purified NuA3 complex identified Sas3 as the catalytic HAT subunit of the complex. Sas3 is the yeast homolog of the human MOZ oncogene. Sas3 is required for both the HAT activity and the integrity of the NuA3 complex. In addition, NuA3 contains the TBP- associated factor, yTAF(II)30, which is also a component of the TFIID, TFIIF, and SWI/SNF complexes. Sas3 mediates interaction of the NuA3 complex with Spt16 both in vivo and in vitro. Spt16 functions as a component of the yeast CP (Cdc68/Pob3) and mammalian FACT (facilitates chromatin transcription) complexes, which are involved in transcription elongation and DNA replication. This interaction suggests that the NuA3 complex might function in concert with FACT-CP to stimulate transcription or replication elongation through nucleosomes by providing a coupled acetyltransferase activity. PMID- 10817756 TI - The steroid receptor coactivator, GRIP-1, is necessary for MEF-2C-dependent gene expression and skeletal muscle differentiation. AB - Nuclear receptor-mediated activation of transcription involves coactivation by cofactors collectively denoted the steroid receptor coactivators (SRCs). The process also involves the subsequent recruitment of p300/CBP and PCAF to a complex that synergistically regulates transcription and remodels the chromatin. PCAF and p300 have also been demonstrated to function as critical coactivators for the muscle-specific basic helix-loop-helix (bHLH) protein MyoD during myogenic commitment. Skeletal muscle differentiation and the activation of muscle specific gene expression is dependent on the concerted action of another bHLH factor, myogenin, and the MADS protein, MEF-2, which function in a cooperative manner. We examined the functional role of one SRC, GRIP-1, in muscle differentiation, an ideal paradigm for the analysis of the determinative events that govern the cell's decision to divide or differentiate. We observed that the mRNA encoding GRIP-1 is expressed in proliferating myoblasts and post-mitotic differentiated myotubes, and that protein levels increase during differentiation. Exogenous/ectopic expression studies with GRIP-1 sense and antisense vectors in myogenic C2C12 cells demonstrated that this SRC is necessary for (1) induction/activation of myogenin, MEF-2, and the crucial cell cycle regulator, p21, and (2) contractile protein expression and myotube formation. Furthermore, we demonstrate that the SRC GRIP-1 coactivates MEF-2C-mediated transcription. GRIP-1 also coactivates the synergistic transactivation of E box-dependent transcription by myogenin and MEF-2C. GST-pulldowns, mammalian two-hybrid analysis, and immunoprecipitation demonstrate that the mechanism involves direct interactions between MEF-2C and GRIP-1 and is associated with the ability of the SRC to interact with the MADS domain of MEF-2C. The HLH region of myogenin mediates the direct interaction of myogenin and GRIP-1. Interestingly, interaction with myogenic factors is mediated by two regions of GRIP-1, an amino terminal bHLH-PAS region and the carboxy-terminal region between amino acids 1158 and 1423 (which encodes an activation domain, has HAT activity, and interacts with the coactivator-associated arginine methyltransferase). This work demonstrates that GRIP-1 potentiates skeletal muscle differentiation by acting as a critical coactivator for MEF-2C-mediated transactivation and is the first study to ascribe a function to the amino-terminal bHLH-PAS region of SRCs. PMID- 10817757 TI - Two protein tyrosine phosphatases, Ptp2 and Ptp3, modulate the subcellular localization of the Hog1 MAP kinase in yeast. AB - The MAP kinase Hog1 transiently accumulates in the nucleus upon activation. Although Hog1 nuclear export correlates with its dephosphorylation, we find that dephosphorylation is not necessary for export. Unexpectedly, a strain lacking the nuclear protein tyrosine phosphatase, Ptp2, showed decreased Hog1 nuclear retention, while a strain lacking the cytoplasmic Ptp3 showed prolonged Hog1 nuclear accumulation, consistent with Ptp2 being a nuclear tether for Hog1 and Ptp3 being a cytoplasmic anchor. In support of this result PTP2 overexpression sequestered Hog1 in the nucleus while PTP3 overexpression restricted Hog1 to the cytoplasm. Thus, Ptp2 and Ptp3 regulate Hog1 localization by binding Hog1. PMID- 10817758 TI - Nucleolin and YB-1 are required for JNK-mediated interleukin-2 mRNA stabilization during T-cell activation. AB - Regulated mRNA turnover is a highly important process, but its mechanism is poorly understood. Using interleukin-2 (IL-2) mRNA as a model, we described a role for the JNK-signaling pathway in stabilization of IL-2 mRNA during T-cell activation, acting via a JNK response element (JRE) in the 5' untranslated region (UTR). We have now identified two major RNA-binding proteins, nucleolin and YB-1, that specifically bind to the JRE. Binding of both proteins is required for IL-2 mRNA stabilization induced by T-cell activation signals and for JNK-induced stabilization in a cell-free system that duplicates essential features of regulated mRNA decay. Nucleolin and YB-1 are required for formation of an IL-2 mRNP complex that responds to specific mRNA stabilizing signals. PMID- 10817759 TI - An evolutionarily conserved RNA stem-loop functions as a sensor that directs feedback regulation of RNase E gene expression. AB - RNase E is a key regulatory enzyme that controls the principal pathway for mRNA degradation in Escherichia coli. The cellular concentration of this endonuclease is governed by a feedback mechanism in which RNase E tightly regulates its own synthesis. Autoregulation is mediated in cis by the 361-nucleotide 5' untranslated region (UTR) of rne (RNase E) mRNA. Here we report the determination of the secondary structure of the rne 5' UTR by phylogenetic comparison and chemical alkylation, together with dissection studies to identify the 5' UTR element that mediates autoregulation. Our findings reveal that the structure and function of the rne 5' UTRs are evolutionarily well conserved despite extensive sequence divergence. Within the rne 5' UTRs are multiple RNA secondary structure elements, two of which function in cis to mediate feedback regulation of rne gene expression. The more potent of these two elements is a stem-loop structure containing an internal loop whose sequence is the most highly conserved of any region of the rne 5' UTR. Our data show that this stem-loop functions as a sensor of cellular RNase E activity that directs autoregulation by modulating the degradation rate of rne mRNA in response to changes in RNase E activity. PMID- 10817760 TI - (CA/GT)(n) microsatellites affect homologous recombination during yeast meiosis. AB - One of the most common microsatellites in eukaryotes consists of tandem arrays of the dinucleotide GT. Although the study of the instability of such repetitive DNA has been extremely fruitful over the last decade, no biological function has been demonstrated for these sequences. We investigated the genetic behavior of a region of the yeast Saccharomyces cerevisiae genome containing a 39-CA/GT dinucleotide repeat sequence. When the microsatellite sequence was present at the ARG4 locus on homologous chromosomes, diploid cells undergoing meiosis generated an excess of tetrads containing a conversion of the region restricted to the region of the microsatellite close to the recombination-initiation double-strand break. Moreover, whereas the repetitive sequence had no effect on the frequency of single crossover, its presence strongly stimulated the formation of multiple crossovers. The combined data strongly suggest that numerous recombination events are restricted to the initiation side of the microsatellite as though progression of the strand exchange initiated at the ARG4 promoter locus was impaired by the repetitive sequence. This observation corroborates in vitro experiments that demonstrated that RecA-promoted strand exchange is inhibited by CA/GT dinucleotide tracts. Surprisingly, meiotic instability of the microsatellite was very high (>0.1 alterations per tetrad) in all the spores with parental and recombinant chromosomes. PMID- 10817761 TI - PAT1, a new member of the GRAS family, is involved in phytochrome A signal transduction. AB - Light signaling via the phytochrome A (phyA) photoreceptor controls basic plant developmental processes including de-etiolation and hypocotyl elongation. We have identified a new Arabidopsis mutant, pat (phytochrome A signal transduction)1-1, which shows strongly reduced responses in continuous far-red light. Physiological and molecular data indicate that this mutant is disrupted at an early step of phyA signal transduction. The PAT1 gene encodes a cytoplasmic protein of 490 amino acids with sequence homologies to the plant-specific GRAS regulatory protein family. In the pat1-1 mutant, a T-DNA insertion introduces a premature stop codon, which likely results in the production of a truncated PAT1 protein of 341 amino acids. The semidominant phenotype of this mutant can be recapitulated by overexpression of an appropriately truncated PAT1 gene in the wild type. The results indicate that the truncated PAT1 protein acts in a dominant-negative fashion to inhibit phyA signaling. PMID- 10817763 TI - Treating fibromyalgia. PMID- 10817764 TI - Secondary prevention of hip fracture. PMID- 10817762 TI - The zebrafish bonnie and clyde gene encodes a Mix family homeodomain protein that regulates the generation of endodermal precursors. AB - Vertebrate endoderm development has recently become the focus of intense investigation. In this report, we first show that the zebrafish bonnie and clyde (bon) gene plays a critical early role in endoderm formation. bon mutants exhibit a profound reduction in the number of sox17-expressing endodermal precursors formed during gastrulation, and, consequently, a profound reduction in gut tissue at later stages. The endodermal precursors that do form in bon mutants, however, appear to differentiate normally indicating that bon is not required at later steps of endoderm development. We further demonstrate that bon encodes a paired class homeodomain protein of the Mix family that is expressed transiently before and during early gastrulation in both mesodermal and endodermal progenitors. Overexpression of bon can rescue endodermal gene expression and the formation of a gut tube in bon mutants. Analysis of a newly identified mutant allele reveals that a single amino acid substitution in the DNA recognition helix of the homeodomain creates a dominant interfering form of Bon when overexpressed. We also show through loss- and gain-of-function analyses that Bon functions exclusively downstream of cyclops and squint signaling. Together, our data demonstrate that Bon is a critical transcriptional regulator of early endoderm formation. PMID- 10817765 TI - Waiting times and referrals. PMID- 10817766 TI - Systemic lupus erythematosus: a model for atherogenesis? PMID- 10817767 TI - Managing seronegative spondarthritides. PMID- 10817768 TI - Does waiting matter? A randomized controlled trial of new non-urgent rheumatology out-patient referrals. AB - OBJECTIVE: To examine the effect of waiting times on the health status of patients referred for a non-urgent rheumatology opinion. METHODS: The study was a randomized controlled clinical study evaluating a 'fast track' appointment with a 6-week target waiting time against an 'ordinary' appointment in the main city out patient clinic of the rheumatology service for the Lothian and Borders region (population approximately 1 million). Health status was measured using the SF12 physical and mental summary component T-scores and pain was measured with a 100 mm visual analogue pain scale. Secondary outcomes were health utility and perceived health both measured with the EuroQol instrument, mental health measured with the Hospital Anxiety and Depression scale, disability with the modified Health Assessment Questionnaire and economic costs measured from a societal perspective. RESULTS: Mean waiting times were 43 days (sigma = +/-16) and 105 days (sigma = +/-51) for 'fast track' and 'ordinary' appointments, respectively. Both groups showed significant improvements in mean [95% confidence interval (CI)] scores for pain: 11 (7, 16)(P < 0.001); physical health status: 4 (2, 5) (P < 0.001); mental health status: 2 (0.1, 4) (P < 0.02); and health utility: 0.11 (0.07, 0.16) (P < 0.001) by the end of the 15-month period of the study, but there was no significant difference between either arm of the study. CONCLUSIONS: Rationing by delay was not detrimental to either mental or physical health and patients in both arms of the study showed significant and similar improvement in health by 15 months. Expenditure of resources on waiting times without regard to clinical outcomes is likely to be wasteful and additional resources should be directed at achieving the greatest clinical benefit. More research into effective methods of controlling demand and better identification of those who would benefit from access to specialist care is needed. PMID- 10817769 TI - Linkage analysis of chromosome 2q in osteoarthritis. AB - BACKGROUND: In independent linkage studies chromosome 2q11-q24 and chromosome 2q23-35 have previously been implicated as regions potentially harbouring susceptibility loci for osteoarthritis (OA). OBJECTIVE: To test chromosome 2q for linkage to idiopathic osteoarthritis. METHODS: Using a cohort of 481 OA families that each contained at least one affected sibling pair with severe end-stage disease (ascertained by hip or knee joint replacement surgery), we conducted a linkage analysis of chromosome 2q using 16 polymorphic microsatellite markers at an average spacing of one marker every 8.5 cM. RESULTS: Our results provide suggestive evidence for a locus at 2q31 with a maximum multipoint logarithm of the odds score (MLS) of 1.22 which increased to 2.19 in those families concordant for hip-only disease (n = 311). This suggestive linkage was greater in male-hip families (MLS = 1.57, n = 71) than in female-hip families (MLS = 0.71, n = 132). CONCLUSIONS: Chromosome 2q is likely to contain at least one susceptibility locus for OA. PMID- 10817770 TI - The reliability of the three-dimensional FASTRAK measurement system in measuring cervical spine and shoulder range of motion in healthy subjects. AB - OBJECTIVES: To assess the inter-observer and intra-observer reliability of a new three-dimensional measurement system, the FASTRAK, in measuring cervical spine flexion/extension, lateral flexion and rotation and shoulder flexion/extension, abduction and external rotation in healthy subjects. METHODS: The study was conducted in two parts. One part assessed inter-observer reliability with two observers measuring 40 subjects. The other part assessed intra-observer reliability with one observer measuring 32 subjects on three occasions. All subjects had unrestricted, pain-free cervical spine and shoulder movement. Reliability was measured by the intraclass correlation coefficient [ICC(2,1)]. RESULTS: The inter-observer ICCs for the cervical spine ranged from 0.61 to 0.89 and for the shoulder from 0.68 to 0.75. After removal of outliers, all ICCs were above 0.70. Intra-observer ICCs for the cervical spine ranged from 0.54 to 0.82 and for the shoulder from 0.62 to 0.81. After removal of outliers, all ICCs were above 0.70 except for shoulder abduction (0.62). CONCLUSIONS: Whilst all movements measured by the FASTRAK showed good reliability, the reliability of the whole movement in a plane (e.g. left plus right lateral flexion) was better than for the separate movements (e.g. left and right lateral flexion taken separately). Inter-observer reliability was generally better than intra-observer reliability for most cervical spine movements, suggesting that variability of movement within subjects (e.g. over a period of days) for these movements was greater than variability between measures on the same occasion. PMID- 10817771 TI - Longitudinal analysis of bone mineral density in pre-menopausal female systemic lupus erythematosus patients: deleterious role of glucocorticoid therapy at the lumbar spine. AB - OBJECTIVE: To evaluate whether bone loss occurs over time in pre-menopausal systemic lupus erythematosus (SLE) patients. METHODS: We performed a longitudinal bone mineral density (BMD) analysis in a group of 35 pre-menopausal female SLE patients. Lumbar spine and hip (total and sub-regions) BMDs were measured twice 21 +/- 11 (mean +/- S.D.) months apart by dual-energy X-ray absorptiometry. RESULTS: In the whole cohort of SLE patients, significant bone loss was observed at the lumbar spine (-1.22%/yr) but not at the total hip. Further analyses indicated that lumbar spine bone loss (-2.12%/yr) occurred exclusively in the subgroup of patients who had taken a mean prednisolone daily dose >7.5 mg between the two BMD measurements. Moreover, bone loss was more important in patients who had previously received a cumulative prednisolone dose 7.5 mg. PMID- 10817772 TI - Major histocompatility complex haplotypic associations in Felty's syndrome and large granular lymphocyte syndrome are secondary to allelic association with HLA DRB1 *0401. AB - OBJECTIVE: To investigate the role of HLA class I in susceptibility to Felty's syndrome (FS) and large granular lymphocyte (LGL) syndrome. METHODS: Fifty caucasoid FS patients, and 55 patients with LGL syndrome, of whom 26 had arthritis and 29 did not, were studied. Complete HLA class I and HLA-DR typing including, where relevant, DRB1*04 subtyping was carried out by molecular methods. Comparison was made with 78 unselected healthy caucasoid controls and a further 29 DRB1*0401+ individuals. RESULTS: A significant association was found between HLA-A*02 and FS [odds ratio (OR) 3.9, 95% confidence interval (95% CI) 1.8-8.4, P = 0.0004]. At the B locus, there was an association between B*44 and LGL with arthritis [OR 3.5 (1.3-9.2), P = 0.01]. For HLA-Cw*0501, there was an association with FS [OR 4 (1. 7-9.2) P = 0.0008]. For both FS and LGL with arthritis, the extended haplotype HLA-A*02;B*44;Cw*0501;DRB1*0401 was significantly associated [OR 9.5 (2.6-35), P = 0.0001; OR 4.6 (1-22.4), P = 0.05, respectively]. There was no association between HLA class I or II and LGL without arthritis. All the allelic and haplotypic associations were lost on comparison with HLA-DRB1*0401+ controls. The strongest HLA association was with HLA DRB1*0401 for FS [OR 27.9 (10.3-75.5), P = 10(-13)], and LGL with arthritis [OR 35.4 (9.6-131. 3), P = 10(-10)]. CONCLUSIONS: The major histocompatibility locus (MHC) associations with FS reported here are due to linkage disequilibrium with HLA-DRB1*0401. LGL syndrome with arthritis shows identical class II associations with FS, although there may be subtle immunogenetic differences between the two in the class I region. One of the extended haplotypes reported in a number of studies for FS and rheumatoid arthritis (summarized as HLA-A*02;Cw*0501; B*44;TNFb5;TNFa6;TNFd4;C4A*3;C4BQ*0;DRB1*0 401;DQB1*0301) is likely to be attributable to strong primary association with HLA-DRB1*0401, rather than to epistatic interaction between these loci. PMID- 10817773 TI - A prospective study of survival and prognostic indicators of systemic lupus erythematosus in a southern Chinese population. AB - OBJECTIVES: To study the survival rate and prognostic indicators of systemic lupus erythematosus (SLE) in a southern Chinese population. METHODS: One hundred and eighty-six patients with SLE diagnosed between 1992 and 1999 were prospectively followed. Clinical features at presentation, subsequent evolving features, autoantibody profile, damage scores and mortality data were obtained. Prognostic factors for survival were studied by statistical analysis. RESULTS: One hundred and sixty-three female and 23 male SLE patients were studied. The female to male ratio was 7.1 to 1 and the mean age at presentation was 33.6 yr (range 12-75). The mean disease duration was 45.2 months. At diagnosis, arthritis, malar rash and alopecia were the commonest features. During follow-up, the prevalence of nephritis, arthritis, photosensitivity and haematological disease increased significantly. Thirty-one per cent of the patients had organ damage at the time of data analysis and renal disease was the commonest cause. Logistic regression revealed that central nervous system disease, discoid lesions and treatment with high-dose steroid were independent predictors for damage. Nine patients died during the study period (three of disease-related complications and six of infections). The 3-, 5-, and 7-yr survival rates of our cohort were 97, 93 and 93%, respectively. Cox regression analysis revealed that thrombocytopenia and high-dose steroid treatment were independent risk factors for mortality. CONCLUSIONS: The survival of SLE in our southern Chinese patients is similar to that of the Caucasian series reported in the 1990s. Although nephritis contributes to organ damage, it is not a major determinant for survival. Infection remains the commonest cause of death. High-dose steroid treatment and thrombocytopenia are independent risk factors for mortality. Judicious use of immunosuppressive agents is necessary to improve the short-term survival of SLE. PMID- 10817774 TI - A 1-year follow-up study of dynamic magnetic resonance imaging in early rheumatoid arthritis reveals synovitis to be increased in shared epitope-positive patients and predictive of erosions at 1 year. AB - OBJECTIVES: Dynamic magnetic resonance imaging (MRI) allows visualization of the synovial membrane and measurement of synovitis within the joint. A cohort of patients with early rheumatoid arthritis (RA) were studied using MRI of the dominant wrist and clinical assessments. Associations between synovitis and the shared epitope genotype (SE) were looked for and synovitis as a predictor of joint erosion was examined. METHODS: Gadolinium-enhanced MRI scans of the dominant wrist were performed in 42 early RA patients at baseline (median disease duration = 4 months) and after 1 yr. Images were obtained at 42-s intervals over the first 6 min after gadolinium-diethylenetriamine pentaacetic acid injection using six cuts in the coronal plane, 2 mm apart. The site of maximal synovial enhancement was selected as the region of interest (ROI). The rate of enhancement (E-rate) was calculated and compared with synovitis scores from static MRI scans, clinical disease activity scores and HLA-DRB1*04/01 genotyping [sequence-specific primer polymerase chain reaction (SSP-PCR) and DNA sequencing]. RESULTS: Reproducibility of the E-rate measurement was assessed by re-evaluating 10 randomly selected scans in a blinded fashion. Intra-observer reliability was high with an intraclass correlation coefficient of 0.91, 95% confidence interval (CI) 0.65-0.97. The E-rate correlated strongly at baseline with the maximum level of synovial enhancement (E-max) (r = 0.88, P < 0.0001) and the static MRI synovitis score (r = 0.52, P = 0.0004). There was also a weaker but significant correlation between E-rate and the pain score (r = 0.29, P = 0.04). The E-rate fell from baseline to 1 yr (P = 0.02) concordant with clinical improvement after treatment with standard therapies. E-rate scores were higher in SE+ than SE - patients (F(1,25) = 5.19, P = 0.03) and were predictive of MRI erosions at 1 yr [chi square = 5.0 (1 d.f.), P = 0.03]. The baseline C-reactive protein (CRP) was also predictive of MRI erosions at 1 yr to a similar degree [chi-square = 4.7 (1 d.f. ), P = 0.03] but the mean static synovitis score at baseline was the strongest predictor [chi-square = 9.2 (1 d.f.), P = 0.003]. CONCLUSIONS: These results show that dynamic MRI can be used to score synovitis objectively in early RA patients. Synovitis was greater in SE+ patients, suggesting an early genetic influence on joint inflammation, and was predictive for the development of erosions at 1 yr. PMID- 10817775 TI - Prevalence of rheumatic manifestations and antineutrophil cytoplasmic antibodies in haematological malignancies. A prospective study. AB - OBJECTIVE: To evaluate the prevalence of antineutrophil cytoplasmic antibodies (ANCA) and rheumatic manifestations associated with chronic haematological malignancies. METHODS: Two groups of patients were prospectively studied (group I: 60 patients with myelodysplastic syndromes and group II: 140 patients with lymphoid malignancies) for clinical 'immune' manifestations and ANCA. RESULTS: In the myelodysplastic group, six patients had ANCA-negative systemic medium-size vasculitis, one had systemic vasculitis with cytoplasmic ANCA, one relapsing polychondritis, one giant cell arteritis, one polymyalgia rheumatica, one polyarthritis and two fasciitis. In group II, two patients had ANCA-negative systemic vasculitis, two had leucocytoclastic vasculitis associated with tuberculosis, two had polyarthritis, one polymyalgia rheumatica and one giant cell arteritis. Six sera were ANCA-positive with perinuclear pattern in four cases, atypical pattern in one and cytoplasmic pattern in one. Two sera had anti myeloperoxidase (MPO) specificity, and others had no known specificity; none had anti-proteinase 3 (PR3) specificity. Global prevalence of ANCA in our cohort was 3%, similar to the French general population. CONCLUSION: Polyarteritis nodosa type systemic vasculitis and polymyalgia rheumatica were the most frequent findings (18%) in myelodysplastic syndromes and particularly in chronic myelomonocytic leukaemia. ANCA were not helpful for the diagnosis of vasculitis. Vasculitis associated with infection, in particular tuberculosis, must be ruled out. PMID- 10817776 TI - Intravenous immunoglobulin therapy of antiphospholipid syndrome. AB - OBJECTIVE: To review the role of intravenous immunoglobulin (IVIg) in antiphospholipid syndrome (APS). METHODS: A literature search was carried out for the immunopathogenesis of APS, laboratory evidence for the beneficial effect of IVIg in APS, and the clinical use of IVIg in APS. RESULTS: There is both laboratory and clinical evidence for the beneficial role of IVIg in APS. IVIg succeeded in in vitro inhibition of anticardiolipin antibodies and lupus anticoagulant, and in the amelioration of experimental APS. Although there are few case reports about IVIg therapy in the haematological manifestations of APS, most of the reports focus on the use of IVIg in the obstetric complications of APS. Hence, in several patient series the use of IVIg either solely or in combination with aspirin/heparin resulted in successful pregnancy outcome in the vast majority of APS patients with recurrent abortions. In addition, IVIg was also beneficial in antiphospholipid antibody-positive patients undergoing in vitro fertilization. CONCLUSION: APS, an autoimmune disease whose main features are vascular thrombosis and pregnancy morbidity, is a good candidate for immunotherapy with IVIg that contains anti-idiotypes directed towards patients' pathogenic antiphospholipid antibodies. Future research should determine when to use anticoagulation, IVIg or both in the treatment of APS. PMID- 10817777 TI - The prevalence of antibodies to anionic phospholipids in patients with the primary antiphospholipid syndrome, systemic lupus erythematosus and their relatives and spouses. AB - OBJECTIVES: Antiphospholipid antibodies (aPL) have been associated with syndromes involving thrombosis, fetal loss and thrombocytopenia. Genetic and environmental conditions are among the factors attributed to the cause of autoimmune diseases such as the antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE). The aim of this study was to determine whether these factors determine the prevalence of aPL. METHODS: Three groups of patients were tested for the presence of IgG, IgM and IgA anticardiolipin (aCL), antiphosphatidylinositol (aPI), antiphosphatidylglycerol (aPG) and antiphosphatidylserine (aPS) antibodies: (i) patients with primary APS (PAPS); (ii) patients with SLE and secondary APS; and (iii) patients with SLE without APS. First-degree relatives and spouses of patients with SLE/APS were also tested for circulating aPL. RESULTS: IgG aPL were particularly prevalent in patients with PAPS. IgG aPI and aCL were more prevalent in patients with PAPS than the IgM equivalents (P < 0.0001). Notably, none of the patients with PAPS had IgA aPL. A significantly higher number of relatives of patients with SLE/APS possessed IgG aPL than the normal controls. Except for aPG (P < 0.03), the prevalence of these antibodies in the relatives was not significantly different from patients with SLE/APS. The relatives also had significantly higher prevalence of IgG aPI, aPS and aCL antibodies than IgM aPL antibodies. In contrast, the prevalence of IgG aPL in the spouses was no different than in the healthy controls. CONCLUSIONS: Genetic factors, shared by patients and their relatives, seem to have some effect on the prevalence of aPL in the subjects studied, while environmental factors shared by spouses appear to have no influence. PMID- 10817778 TI - Tumour necrosis factor alpha and its soluble receptors in juvenile chronic arthritis. AB - OBJECTIVE: To identify possible imbalance of tumour necrosis factor alpha (TNFalpha) and its soluble receptors in the different subgroups of juvenile chronic arthritis (JCA). METHODS: Serum and synovial fluid samples from 45 children were examined, 25 pauciarticular JCA, 13 polyarticular JCA and seven spondyloarthropathy. TNFalpha, sTNFRI and sTNFRII levels were measured by EASIA and enzyme-linked immunosorbent assay (ELISA). Analysis of the results was carried out using non-parametric tests: Kruskal-Wallis one-way analysis of variance was used to compare the three clinical subgroups; the Mann-Whitney U test was used to compare group medians. RESULTS: Thirty-three serum samples were assayed for TNFalpha. There was no significant difference between the three groups using the Kruskal-Wallis analysis of variance. Analysis of synovial fluid TNF levels showed significantly lower levels in the spondyloarthropathy group compared with the pauciarticular JCA (P = 0.01) and the polyarticular group (P = 0.002). Significantly higher levels of sTNFRI were observed in the synovial fluid of the polyarticular JCA group compared with the pauciarticular JCA group (P = 0.004) and similarly for sTNFRII (P = 0.03). Molar ratios were calculated for TNF vs sTNFRI. The sTNFRI/TNFalpha ratio was significantly higher in the spondyloarthropathy group compared with the pauci- (P 0.003) and the polyarticular JCA subgroups (P = 0.003). The combined soluble receptor levels expressed as molar ratio to TNF again showed a significantly higher ratio in the spondyloarthropathy group compared with the pauciarticular group (P = 0.01) and compared with the polyarticular group (P = 0.05). CONCLUSION: These results suggest that the increased joint destruction observed in polyarticular disease compared with the other two subtypes may be related to the lower sTNFR/TNFalpha ratios observed. PMID- 10817779 TI - Reflex sympathetic dystrophy in hypophosphataemic osteomalacia with femoral neck fracture: a case report. AB - We report a male patient who presented with suspicion of skeletal metastases based upon an abnormal 99-mTc bone scan, which showed increased uptake at both femoral heads, left femoral neck, and several ribs. The images also suggested reflex sympathetic dystrophy, subcapital fracture of the left femur, and rib fractures. A diagnosis of hypophosphataemic osteomalacia was finally made. PMID- 10817780 TI - Buerger's disease (thromboangiitis obliterans): a reversible cause of upper limb digital infarcts. PMID- 10817781 TI - Giant geode (pseudocyst) formation of the femoral neck in a case of osteoarthritis. PMID- 10817782 TI - Susceptibility to ankylosing spondylitis. PMID- 10817783 TI - Long-standing and intractable ascites involved in renal vein thrombosis of a patient with systemic lupus erythematosus. PMID- 10817784 TI - A case of hypopituitarism associated with antiphospholipid syndrome. PMID- 10817786 TI - Forthcoming events PMID- 10817785 TI - Use of an electronic monitoring aid to investigate the medication pattern of analgesics and non-steroidal anti-inflammatory drugs prescribed for osteoarthritis. PMID- 10817787 TI - Atopy, asthma, and the mycobacteria. PMID- 10817788 TI - Secondary genetic factors in cystic fibrosis lung disease. PMID- 10817789 TI - Patient accrual into chemotherapy trials in non-small cell lung cancer. PMID- 10817790 TI - International patterns of tuberculosis and the prevalence of symptoms of asthma, rhinitis, and eczema. AB - BACKGROUND: An ecological analysis was conducted of the relationship between tuberculosis notification rates and the prevalence of symptoms of asthma, allergic rhinoconjunctivitis, and atopic eczema in 85 centres from 23 countries in which standardised data are available. These essentially comprised countries in Europe as well as the USA, Canada, Australia, and New Zealand. METHODS: Tuberculosis notification rates were obtained from the World Health Organization. Data on the prevalence of symptoms of asthma, rhinitis, and eczema in 235 477 children aged 13-14 years were based on the responses to the written and video questionnaires from the International Study of Asthma and Allergies in Childhood (ISAAC). The analysis was adjusted for gross national product (GNP) as an estimate of the level of affluence. RESULTS: Tuberculosis notification rates were significantly inversely associated with the lifetime prevalence of wheeze and asthma and the 12 month period prevalence of wheeze at rest as assessed by the video questionnaire. An increase in the tuberculosis notification rates of 25 per 100 000 was associated with an absolute decrease in the prevalence of wheeze ever of 4.7%. Symptoms of allergic rhinoconjunctivitis in the past 12 months were inversely associated with tuberculosis notification rates, but there were no other significant associations with other ISAAC questions on allergic rhinoconjunctivitis or atopic eczema. CONCLUSIONS: These findings are consistent with recent experimental evidence which suggests that exposure to Mycobacterium tuberculosis may reduce the risk of developing asthma. PMID- 10817791 TI - Absence of relationship between tuberculin reactivity and atopy in BCG vaccinated young adults. AB - BACKGROUND: An inverse association between tuberculin responses and atopy has been observed in Japanese children, indicating that BCG immunisation, subclinical exposure to Mycobacterium tuberculosis without clinical disease, or host characteristics may influence the T helper (Th) lymphocyte balance with decreased atopy as a result. This study was undertaken to determine whether tuberculin reactivity is inversely related to atopy in young adults vaccinated with BCG at the age of 14. METHODS: Men and women aged 20-44 years were tested using the adrenaline-Pirquet test with Norwegian produced synthetic medium tuberculin (n = 891). In addition, their serum total and specific IgE antibodies against mite, cat, timothy grass, mould and birch were measured. RESULTS: Of the 574 subjects with complete examinations, 64% had a positive adrenaline-Pirquet tuberculin test (> or =4 mm) and 27% exhibited IgE antibodies (> or =0.35 kU/l) to one or more of the five specific allergens. The geometric mean of total serum IgE in the population was 30.2 kU/l. Tuberculin reactivity and log IgE were not correlated (r = 0.043, p = 0.30). The mean tuberculin reactivity was 4.6 mm, 4.9 mm, and 5.0 mm in the lower, middle and upper tertile of IgE distribution (<14 kU/l, 14-61 kU/l, >61 kU/l). The prevalence of atopy, as assessed by either the presence of any of the five specific IgE antibodies or by each specific IgE antibody separately, did not differ between subjects with a positive and those with a negative tuberculin test. These results persisted after adjustment for age, sex, and smoking status in multivariate logistic regression analyses. CONCLUSIONS: In this young adult population, BCG vaccinated at the age of 14, no significant relationship between a positive tuberculin reaction and atopy was observed. If a true relationship had been found, our study suggests that it may be limited to populations immunised in early childhood when a substantial modulation of the immune system can occur. PMID- 10817792 TI - TGF-beta(1) genotype and accelerated decline in lung function of patients with cystic fibrosis. AB - BACKGROUND: Polymorphisms in transforming growth factor (TGF)-beta(1) associated with variations in cytokine levels are linked to fibrosis in a number of tissues. However, the contribution of this cytokine to organ fibrosis in patients with cystic fibrosis is presently unclear. This study was undertaken to examine the association between TGF-beta(1) gene polymorphisms and the development of pulmonary dysfunction in patients with cystic fibrosis. METHODS: Polymorphisms in the TGF-beta(1) gene defining amino acids of codons 10 and 25 were determined by ARMS-PCR using DNA stored on 171 Caucasian patients who were homozygous for the DeltaF508 mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Clinical information on the patients was obtained from medical records. RESULTS: Patients with cystic fibrosis of a TGF-beta(1) high producer genotype for codon 10 had more rapid deterioration in lung function than those with a TGF-beta(1) low producer genotype. The relative risk of accelerated decline in forced expiratory volume in one second (FEV(1)) to 50% predicted and forced vital capacity (FVC) to 70% predicted of patients with a high producer genotype was 1.74 (95% CI 1.11 to 2. 73) compared with 1.95 (95% CI 1.24 to 3.06) for those with a low producer genotype. DISCUSSION: TGF-beta(1) genotypes may have a role in mediating pulmonary dysfunction in patients with cystic fibrosis. Further work is required to determine whether inhibition of TGF-beta(1) activity in these patients may slow disease progression. PMID- 10817793 TI - Recruitment of patients with lung cancer into a randomised clinical trial: experience at two centres. On behalf of the Big Lung Trial Steering Committee. AB - BACKGROUND: The entry of patients into randomised clinical trials (RCTs) in lung cancer is low. A study was undertaken to assess the reasons why patients with non small cell lung cancer did not enter a trial involving randomisation to receive or not receive three courses of cisplatin based chemotherapy in addition to primary treatment by surgery, radiotherapy, or best supportive care. METHODS: The study was carried out in two large London institutions with a special interest in recruiting patients to lung cancer trials. Patients recently diagnosed as having non-small cell lung cancer were prospectively identified and followed to see whether they entered the RCT and, if not, to identify the main reasons why. RESULTS: Six hundred and eighty eight patients newly diagnosed with non-small cell lung cancer were identified between November 1995 and July 1998; 274 (39.8%) were deemed ineligible for the RCT for clinical reasons, most frequently their general condition rendering them unfit for chemotherapy. Another 161 (23.4%) were ineligible for logistical reasons-for example, they were discharged to centres not participating in the RCT or they were not considered for the trial at an appropriate time in their management. Of 253 potentially eligible patients, only 63 (24.9% of those eligible) agreed to enter the RCT and four entered another study. Of those who did not enter, 77 (41.4%) declined without stating a reason, 61 (32.8%) did not want chemotherapy, and only eight (4.3%) expressed a wish to have chemotherapy. CONCLUSIONS: Despite considerable time and effort, the proportion of patients recruited was small (9.2%). Many seen were ineligible but, of 253 potentially eligible patients, 186 (73.5%) refused to enter the RCT. PMID- 10817794 TI - Asthma aggravation, combustion, and stagnant air. AB - BACKGROUND: The relationship between current concentrations of ambient air pollution and adverse health effects is controversial. We report a meteorological index of air stagnation that is associated with daily visits to the emergency department for asthma in two urban areas. METHODS: Data on daily values of a stagnation persistence index and visits to the emergency department for asthma were collected for approximately two years in Spokane, Washington, USA and for 15 months in Seattle, Washington, USA. The stagnation persistence index represents the number of hours during the 24 hour day when surface wind speeds are less than the annual hourly median value, an index readily available for most urban areas. Associations between the daily stagnation persistence index and daily emergency department visits for asthma were tested using a generalised additive Poisson regression model. A factor analysis of particulate matter (PM(2.5)) composition was performed to identify the pollutants associated with increased asthma visits. RESULTS: The relative rate of the association between a visit to the emergency department for asthma and the stagnation persistence index was 1. 12 (95% CI 1.05 to 1.19) in Spokane and 1.21 (95% CI 1.09 to 1.35) in Seattle for an increase of 11 and 10 hours, respectively, of low wind speed in a given day. The stagnation persistence index was only correlated with one set of factor loadings; that cluster included the stagnation persistence index, carbon monoxide, and organic/elemental carbon. CONCLUSION: Increased air stagnation was shown to be a surrogate for accumulation of the products of incomplete combustion, including carbon monoxide and fine particulate levels of organic and elemental carbon, and was more strongly associated with asthma aggravation than any one of the measured pollutants. PMID- 10817795 TI - Automated quantitation of circulating neutrophil and eosinophil activation in asthmatic patients. AB - BACKGROUND: Asthma has been associated with eosinophil activation, measured in serum, sputum, bronchoalveolar lavage (BAL) fluid, and urine. A whole blood automated method was developed to assess eosinophil and neutrophil activity in terms of peroxidase content and cell morphology using the Bayer haematology analyser. The method was applied to an in vitro stimulation model when fMLP was added to whole blood and the samples were then analysed for changes in granularity and shape. In addition, cells stimulated with interleukin (IL)-8 were examined by electron microscopy. METHODS: A cross sectional analysis was performed on venous blood from non-atopic, non-asthmatic normal subjects (n = 37), mild (n = 46) and symptomatic (n = 22) asthmatic patients on inhaled beta(2) agonist only, and more severe asthmatic patients (n = 17) on inhaled and oral corticosteroid therapy. Samples were analysed by the haematology analyser and peroxidase leucograms gated using the WinMDI software program. RESULTS: There were significant differences in the amount of light scatter by the neutrophil populations in the symptomatic (p = 0.007) and severe asthmatic (p = 0.0001) groups compared with the control group. However, abnormalities in eosinophil populations were not observed. In vitro activation of whole blood with fMLP caused similar changes in neutrophil light scatter, suggesting that neutrophil activation is present in peripheral blood of symptomatic asthmatic patients. IL-8 caused a change in shape of the neutrophils seen using transmission electron microscopy. CONCLUSIONS: Evidence of neutrophil activation can be seen in whole blood from patients with asthma using a novel automated method. This may potentially be applied to other inflammatory diseases. PMID- 10817796 TI - Effect of leukotriene receptor antagonist therapy on the risk of asthma exacerbations in patients with mild to moderate asthma: an integrated analysis of zafirlukast trials. AB - BACKGROUND: Asthma exacerbations contribute substantially to morbidity, and their reduction is an important therapeutic objective. In this integrated analysis the risk of asthma exacerbations was assessed during treatment with the leukotriene receptor antagonist zafirlukast. METHODS: Data were collected from all five double blind, multicentre, randomised, placebo controlled, 13 week trials of zafirlukast 20 mg twice daily performed in steroid-naive patients with mild to moderate asthma. Exacerbation data were collected prospectively during monitoring of adverse events and concomitant medication use. Pooled data were used to assess the relative risk of asthma exacerbations using three definitions: worsening of asthma leading to withdrawal from the study; requirement for additional anti asthma therapy (excluding increased short acting beta(2) agonist use); and requirement for oral corticosteroid therapy. RESULTS: The proportion of patients with an asthma exacerbation leading to withdrawal was consistently lower in the group treated with zafirlukast 20 mg twice daily than in the placebo group. Overall, the risk of an asthma exacerbation requiring withdrawal from zafirlukast therapy was approximately half that of placebo (odds ratio 0.45; 95% CI 0.26 to 0.76; p = 0.003). Similar results were observed for exacerbations requiring additional control medication (odds ratio = 0.47; 95% CI 0.30 to 0.74; p = 0.001) and oral corticosteroid rescue (odds ratio = 0.53; 95% CI 0.32 to 0.86; p = 0.010). CONCLUSIONS: Zafirlukast in a dose of 20 mg twice daily reduces the risk of asthma exacerbations and the need for additional anti-asthma therapies, fulfilling an important goal of control medication in patients with mild to moderate asthma. PMID- 10817797 TI - Dissociation between exhaled nitric oxide and hyperresponsiveness in children with mild intermittent asthma. AB - BACKGROUND: Bronchial hyperresponsiveness and airway inflammation are distinctive features of asthma. Evaluation of nitric oxide (NO) levels in expired air have been proposed as a reliable method for assessing the airway inflammatory events in asthmatic subjects. A study was undertaken to evaluate whether airway hyperresponsiveness is related to levels of exhaled NO. METHODS: Thirty two steroid-naive atopic children with mild intermittent asthma of mean (SD) age 11.8 (2.3) years and 28 age matched healthy controls were studied to investigate whether baseline lung function or airway hyperresponsiveness is related to levels of exhaled NO. Airway responsiveness was assessed as the dose of methacholine causing a 20% decrease in forced expiratory volume in one second (FEV(1)) from control (PD(20) methacholine) and exhaled NO levels were measured by chemiluminescence analysis of exhaled air. RESULTS: At baseline asthmatic children had significantly higher NO levels than controls (mean difference 25.87 ppb (95% CI 18.91 to 32.83); p<0.0001) but there were no significant differences in lung function parameters (forced vital capacity (FVC), FEV(1) (% pred), and forced expiratory flows at 25-75% of vital capacity (FEF(25-75%))). In the asthmatic group exhaled NO levels were not significantly correlated with baseline lung function values or PD(20) methacholine. CONCLUSIONS: These results suggest that levels of exhaled NO are not accurate predictors of the degree of airway responsiveness to inhaled methacholine in children with mild intermittent asthma. PMID- 10817798 TI - An animal model for allergic penicilliosis induced by the intranasal instillation of viable Penicillium chrysogenum conidia. AB - BACKGROUND: A study was undertaken to determine the consequences of long term intranasal instillation of Penicillium chrysogenum propagules in a mouse model. METHODS: C57 Black/6 mice were inoculated intranasally each week for six weeks with 10(4) viable and non-viable P chrysogenum conidia. Cytokine levels and cellular responses in these animals were then measured. RESULTS: Compared with controls, mice inoculated intranasally each week for six weeks with 10(4) P chrysogenum conidia (average viability 25%) produced significantly more total serum IgE (mean difference 1823.11, lower and upper 95% confidence intervals (CI) 539.09 to 3107.13), peripheral eosinophils (mean difference 5.11, 95% CI 2.24 to 7.99), and airway eosinophilia (rank difference 11.33, 95% CI 9.0 to 20.0). With the exception of airway neutrophilia (mean difference 20.89, 95% CI 3.72 to 38.06), mice inoculated intranasally with 10(4) non-viable conidia did not show significant changes in total serum IgE, peripheral or airway eosinophils. However, when compared with controls, this group (10(4) non-viable) had a significant increase in total serum IgG(2a) (mean difference 1990.56, 95% CI 790.48 to 3190.63) and bronchoalveolar lavage (BAL) fluid levels of interferon (IFN)-gamma (mean difference 274.72, 95% CI 245.26 to 304.19). In addition, lung lavages from mice inoculated intranasally with 10(4) viable P chrysogenum conidia had significantly increased levels of interleukin (IL)-4 (mean difference 285.28, 95% CI 108.73 to 461.82) and IL-5 (mean difference 16.61, 95% CI 11.23 to 21.99). The IgG(2a)/IgE ratio and the IFN-gamma/IL-4 ratio was lower in the group of mice inoculated intranasally with 10(4) viable conidia than in the 10(4) non-viable conidia group and the controls. When proteins were extracted from P chrysogenum conidia, attached to microtitre plates and incubated with serum from the 10(4) viable group, significant increases in conidia-specific IgE and IgG(1) were observed compared with controls, while serum from the 10(4) non-viable group was similar to controls. CONCLUSIONS: These data suggest that long term inhalation of viable P chrysogenum propagules induces type 2 T helper cell mediated (Th2) inflammatory responses such as increases in total and conidia-specific serum IgE and IgG(1), together with BAL fluid levels of IL-4 and IL-5 and peripheral and airway eosinophilia, which are mediators of allergic reactions. PMID- 10817799 TI - Effect of digoxin on global respiratory muscle strength after cholecystectomy: a double blind study. AB - BACKGROUND: Upper abdominal surgery has been shown to impair the function of the respiratory muscles. In addition, controversial results have been reported concerning the effect of digoxin on the diaphragm. The aim of this study was to investigate further the mechanism(s) of respiratory muscle dysfunction after cholecystectomy and the effect of digoxin on the impaired respiratory muscle function. METHODS: Twenty three patients (four men) were studied before and 48 hours after surgery. Eleven received digoxin and 12 placebo. Respiratory muscle strength was assessed 48 hours after surgery by measuring mouth pressure during maximum static inspiratory (PImax) and expiratory (PEmax) efforts before and after 90 minutes of intravenous administration of 0.25 mg digoxin in a double blind, placebo controlled fashion. In addition, spirometric and pain measurements were performed. RESULTS: Postoperatively (+48 h) PImax and PEmax decreased significantly (p<0.01) from their preoperative values in both groups by a similar degree. After administration of digoxin or placebo only the digoxin group showed a significant increase in both PImax (p<0.02) and PEmax (p<0.05) with a mean increase of 15% for PImax and 12.3% for PEmax. The mean difference in PImax (DeltaPImax) and PEmax (DeltaPEmax) between the digoxin and placebo groups was 1.01 (95% CI 0.28 to 2.2) and 1.05 (95% CI 0.04 to 2.4), respectively. Estimates of postoperative pain did not differ between the two groups. Spirometric indices showed a similar restrictive defect postoperatively in both groups but did not change after digoxin or placebo. CONCLUSION: Digoxin improves the impaired global strength of the inspiratory and expiratory muscles after cholecystectomy and this may be clinically relevant. Muscle contractility could play a part in this impairment. PMID- 10817800 TI - Small vessel vasculitis of the lung. PMID- 10817801 TI - A persistent challenge: the diagnosis of respiratory disease in the non-AIDS immunocompromised host. PMID- 10817803 TI - Paradoxical vocal cord motion causing stridor after thyroidectomy. AB - Two women developed stridor immediately after thyroidectomy as a result of paradoxical vocal cord motion. In both cases the cord function showed a normal pattern during vocalisation but paradoxical movement was seen at laryngoscopy during tidal breathing. The abnormality improved in both patients over time with speech therapy. Whilst the syndrome of paradoxical vocal cord motion is classically thought to have a largely psychological aetiology, subtle interference with laryngeal innervation at surgery is more likely to have been the cause in these cases. PMID- 10817804 TI - Lung function tests: physiological principles and clinical applications PMID- 10817802 TI - Indoor air pollution in developing countries and acute lower respiratory infections in children. AB - BACKGROUND: A critical review was conducted of the quantitative literature linking indoor air pollution from household use of biomass fuels with acute respiratory infections in young children, which is focused on, but not confined to, acute lower respiratory infection and pneumonia in children under two years in less developed countries. Biomass in the form of wood, crop residues, and animal dung is used in more than two fifths of the world's households as the principal fuel. METHODS: Medline and other electronic databases were used, but it was also necessary to secure literature from colleagues in less developed countries where not all publications are yet internationally indexed. RESULTS: The studies of indoor air pollution from household biomass fuels are reasonably consistent and, as a group, show a strong significant increase in risk for exposed young children compared with those living in households using cleaner fuels or being otherwise less exposed. Not all studies were able to adjust for confounders, but most of those that did so found that strong and significant risks remained. CONCLUSIONS: It seems that the relative risks are likely to be significant for the exposures considered here. Since acute lower respiratory infection is the chief cause of death in children in less developed countries, and exacts a larger burden of disease than any other disease category for the world population, even small additional risks due to such a ubiquitous exposure as air pollution have important public health implications. In the case of indoor air pollution in households using biomass fuels, the risks also seem to be fairly strong, presumably because of the high daily concentrations of pollutants found in such settings and the large amount of time young children spend with their mothers doing household cooking. Given the large vulnerable populations at risk, there is an urgent need to conduct randomised trials to increase confidence in the cause-effect relationship, to quantify the risk more precisely, to determine the degree of reduction in exposure required to significantly improve health, and to establish the effectiveness of interventions. PMID- 10817805 TI - Aging of the vascular-wall and atherosclerosis. AB - The rapid increase of life expectancy during the last half of the 20th century is changing the expression of cardiovascular disease and of its risk factors. These findings are examined by the separate consideration of atheromatous plaque formation and vascular wall stiffening, known as arteriosclerosis. in humans, these processes may progress together, but in some other species as the rat, only vascular wall stiffening is observed. A saturated fat- and cholesterol-rich diet produces the early appearance of lipidic plaques, which progress to fibrous, sometimes ulcerated, thrombotic lesions. This progression is age dependent; the establishment of lipidic plaques is not. Vascular wall stiffening, characterized by an increase of the collagen-elastin ratio and diffuse deposition of calcium and lipids is also age dependent (arteriosclerosis). Although hyperlipidemia appears to be involved both in plaque formation and wall thickness progression, the detailed mechanisms are not identical. In the oldest age group (above 80 years and in centenarians), high cholesterol values may not be a risk factor as in younger individuals. Among the cellular and molecular mechanisms involved, immune factors and modifications in receptor coupling appear to play a major role. These mechanisms are described in some detail. PMID- 10817806 TI - Age-related changes in vascular responses. AB - Aging causes changes in the structure and function of the vessels that leads to an increase in the incidence of certain cardiovascular diseases, such as hypertension, coronary artery disease, heart failure, and postural hypotension with enhancement of both morbidity and mortality. When aging is associated with hypertension, these changes are increased. Aging alters endothelial cells, and so the vascular tone regulation, reducing the endothelium-dependent relaxations, probably by a decrease in endothelial synthesis or release of nitric oxide. In addition, endothelium-independent relaxations are essentially unaltered, those elicited by beta-adrenoceptor agonists being usually reduced. Aging scarcely modifies the contractions induced by different agents, such as 5 hydroxytryptamine, histamine, high potassium, and angiotensin, whereas reduces those elicited by noradrenaline or endothelin. Vascular Ca(2+) homeostasis appears to be altered in aging. The extracellular Ca(2+) dependence of contractile responses elicited by agonists is enhanced, which explains the increased sensitivity to Ca(2+) antagonists in elderly. Finally, Na(+) pump activity, that controls cellular ionic homeostasis, seems to be reduced in aging. The contractions elicited by Na(+) pump inhibition with ouabain are negatively modulated by the release of a diffusible endothelial factor, an effect lost in aging, being replaced by an endothelium-dependent contracting factor that facilitates ouabain responses. PMID- 10817807 TI - Collagens and atherosclerosis. AB - Smooth muscle cells in the atherosclerotic lesions of diseased arteries produce new extracellular matrix, largely collagenous in nature, which is responsible in part for the occlusion of the vessel lumen by the atherosclerotic plaque. These smooth muscle cells express a different phenotype, responsive to growth factors, to that of the differentiated, nondividing contractile cell in the media. Specific collagens may be involved in the regulation of phenotype and in the migration of the cells to the site of lesion growth. Collagens may also be involved in the calcification of lesions, in the retention of low-density lipoprotein in the vessel wall and in smooth muscle cell survival. Glycation of collagen may promote atherogenesis. Effects as summarized in this short review, are not always, at first sight, consistent. The following points should be kept in mind, though, when considering the response of a cell to collagen. Any effect may be governed not just by the identity of the collagen type as such but by its state of polymerization: monomeric collagen, for instance, whether in solution or immobilized on plastic, may express different effects to the same collagen type when presented in its native polymerized state, e.g., as fibers. The precise identity of the cell and its location may be important: SMCs in secondary culture may not necessarily respond to any given collagen exactly as SMCs within the lesion or possess precisely the same properties, albeit both types are regarded as expressing the same (synthetic) phenotype. Effects may not necessarily be directly attributable to collagen, but to some other matrix constituent bound to collagen. PMID- 10817808 TI - Lipoprotein modification and atherosclerosis in aging. AB - The development of extensive atherosclerosis of major arteries of the heart, brain, and lower extremities is a particularly frequent problem in elderly individuals and is responsible for the majority of the cardiovascular morbidity and mortality in this population. Although the frequency and severity of this problem is well recognized, there has been relatively little investigation of the effects of aging on the development of atherosclerosis. Work by a number of investigators over the last 10-15 years has demonstrated that modifications of lipoproteins, resulting from oxidative stress, glycoxidation, formation of AGE, or other processes may play an important role in atherosclerosis. As described in this review, the aging process may enhance lipoprotein modification and atherosclerosis in several ways. Conditions such as hypertension, diabetes, and menopause all increase in frequency with advancing age and may contribute both directly and indirectly to lipoprotein modification and vascular injury.Additionally, in some studies of older animals and humans, there seems to be evidence for greater in vivo oxidative stress. Whether this is a specific consequence of aging and associated medical conditions, or related to differences in dietary fatty acid or antioxidant content or other lifestyle differences is currently unknown. One important consequence of this may be enhanced susceptibility of lipoproteins to oxidation. Additional study of lipoprotein modifications associated with aging is clearly needed, and may provide new insight and solutions to the common problem of atherosclerosis in the elderly. PMID- 10817809 TI - The protective role of high-density lipoproteins in atherosclerosis. AB - Serum high-density lipoprotein level is known to be correlated inversely with the incidence and mortality rates of ischemic heart disease. Although some reports pointed out that in case of hyperalphalipoproteinemia, lesions in the coronary arteries were occasionally found, it is also noticed that in very rare condition, no atheromatous lesions found even in patients with hereditary alphalipoprotein deficiency (Funke et al., 1991). However, clinical surveys have confirmed that high high-density-lipoprotein cholesterol level is favorable in preventing the development of atheroclerotic lesion and high-density lipoprotein together with apolipoprotein AI are currently considered to be the most reliable parameters in predicting the development of atherosclerosis in hyperlipidemia. PMID- 10817810 TI - Age-related changes in the signaling and function of vascular smooth muscle cells. AB - Aging is an independent risk factor for the development of atherosclerosis, a vascular abnormality that plays a significant role in the development of many cardiovascular disorders. Animal experiments have demonstrated that aging predisposes the vasculature to advanced atherosclerotic disease and vessel injury and that this predisposition is a function of age-associated changes in the vessel wall itself. Because vascular smooth muscle cells play important roles in the pathogenesis of many vascular disorders, identifying age-associated differences in the way these cells respond to extracellular clues has been an area of active research. Currently, the most remarkable differences in intracellular signaling between vascular smooth muscle cells isolated from young and old animals are related to the control of cell migration through the CamKII pathways and the accelerated transition of older vascular smooth muscle cells from the contractile to the synthetic phenotype. These differences may be due to alternative signaling pathways revealed by the inability of older cells to respond to inhibitors, such as transforming growth factor (TGF)-beta1, or to altered interactions with the extracellular matrix resulting from age-associated shifts in integrin expression or changes in the matrix composition of blood vessels. The exact role that these alterations have in explaining age-associated differences in the response of the vessel wall to injury and its increased susceptibility to developing advanced atherosclerotic lesions remains to be determined but will be guided by studies on intracellular signaling mechanisms. PMID- 10817811 TI - Atherosclerosis--an autoimmune disease. AB - Immune-inflammatory processes are increasingly discussed as possible pathogenetic factors involved in the development of atherosclerosis. Here, we summarize data on which we have built our "immunological" hypothesis of atherogenesis. This concept is based on the observation that nearly everybody shows protective cellular and humoral immune reactions against microbial heat shock protein 60 (HSP 60). Because a high degree of antigenic homology exists between microbial (viral, bacterial, parasitic) and human HSP 60, this protective immunity may have to be "paid for" by the danger of cross-reactivity with human HSP 60 that is expressed by endothelial cells of stressed arteries. Arterial endothelial cells are more prone to produce HSP 60 and various adhesion molecules upon exposure to stress factors, including classical risk factors for atherosclerosis, due to their life-long exposure to the high arterial as compared to venous blood pressure. Also, endothelial cells are the first potential targets encountered by circulating HSP 60-specific T cells or antibodies. This concept not only opens new avenues for diagnostic approaches, but also may form the basis for new ways of therapeutic intervention. PMID- 10817813 TI - Studies on the modeling and simulation of a sequential bienzymatic reaction system immobilized in emulsion liquid membrane. AB - Experiments have been carried out to study the reaction engineering behavior of the liquid membrane-encapsulated, sequential bienzymatic reaction system, n 2n glucose. A dynamic mathematical model, free from adjustable parameters, has been developed taking into account peri-emulsion mass transfer, intra-emulsion diffusion, membrane-related mass transfer limitations and substrate and product inhibitions. A finite difference-based, user-friendly software has been developed to solve the model equations. Experimental data satisfactorily correlate with the model. While it is understood that study of sequential bienzymatic reaction system immobilized in emulsion liquid is essential for their industrial exploitation, reaction engineering behavior of such a system in presence of both substrate and product inhibitions has not yet been reported in the literature. Therefore, the model predictions of the present investigations are expected to pave the way for scale-up and design of industrial bioreactors in this field. PMID- 10817812 TI - Gene therapy for angiogenesis, restenosis and related diseases. AB - Gene therapy may be useful for the treatment of atherosclerosis and related diseases. Gene transfer to vascular system can be performed both via intravascular and extravascular routes. Gene transfer to other tissues, such as liver and muscle, can also be used. The first clinical trials for the induction of therapeutic angiogenesis with VEGF gene transfer are under way, and preliminary results are promising. In the prevention of restenosis genes inhibiting cellular proliferation and increasing NO production, such as NOS and VEGF, have been used. However, more basic research is needed to fully understand pathophysiological mechanisms involved in conditions related to atherosclerosis. Also, further developments in gene transfer vectors and gene delivery techniques are required for the improvement of the efficacy of gene therapy. PMID- 10817814 TI - Optimizing initial plasmid copy number distribution for improved protein activity in a recombinant fermentation. AB - Recombinant bacterial cells in a fermentation broth rarely contain the same number of plasmids, even though this simplification is often used. Recent work has however indicated limitations of the simplified approach. Based on these studies, the distribution of plasmid copy numbers per cell has been represented macroscopically here in a Gaussian form for the fraction of biomass as a function of the copy number. Applying this distribution and an experimentally validated kinetic model to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) synthesis by Escherichia coli containing the plasmid pBR Eco gap, it is seen that GAPDH production in a batch fermentation is maximized by a particular initial (non zero) copy number variance and an optimal duration. To implement this distribution in a bioreactor, it is suggested that the profile may be discretized, inocula corresponding to the mean copy number of each fraction prepared, and then combined to obtain the seed culture. PMID- 10817815 TI - Agitation induced mycelial fragmentation of Aspergillus oryzae and Penicillium chrysogenum. AB - Given the impact of mycelial morphology on fermentation performance, it is important to understand the factors that influence it, including agitation induced fragmentation. The successful application of the energy dissipation/circulation function (EDC) to correlate fragmentation of Penicillium chrysogenum with agitation intensity and with different impeller types [5] has already been demonstrated. The EDC function takes into account the specific energy dissipation rate in the impeller swept volume and the frequency of mycelial circulation through that volume. In order to explore whether the EDC function can be used more generally to correlate fragmentation of different filamentous species, the present study extended the concept to agitation-induced, off-line fragmentation of Aspergillus oryzae grown in chemostat culture. The work shows that at EDC values off-line greater than that in the chemostat, fragmentation with different impellers can be correlated with the EDC. For EDC values less than those used in the chemostat, fragmentation did not occur. The earlier results of Justen et al. [5] with Penicillium chrysogenum are also reconsidered and found to behave similarly. PMID- 10817816 TI - alpha-Amylase production by Bacillus subtilis with dregs in an external-loop airlift bioreactor. AB - An external-loop airlift bioreactor, with a low ratio 2.9 of height-to-diameter of the riser and a ratio 6.6 of riser-to-downcomer diameter, was used to produce alpha-amylase from fermentation with dregs by Bacillus subtilis. The effects of gas flow rate and liquid volume on alpha-amylase production were investigated. After a 36-h fermentation time, an average of 432.3U/ml alpha-amylase activity was obtained under the conditions of liquid volume 8.5l and gas flow rate 1.2vvm for the first 12h of fermentation, 1.4vvm from 12 to 27h, and 1.2vvm from 27h to the end. The activity was higher than that obtained in shaking flasks (409.0U/ml) and in a mechanically stirred tank bioreactor (397.2U/ml) under optimized operating conditions. The fermentation cycle of the airlift bioreactor was shorter than the 48h required for the shaking flasks and close to the 36h of the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the traditional mechanically stirred tank bioreactor to produce alpha-amylase from fermentation by Bacillus subtilis with dregs. PMID- 10817817 TI - A study on the surface hydrophobicity of lipases. AB - Surface properties, including surface net and local hydrophobicities, of bovine serum albumin, gamma-globulin, and six lipases of different origins were evaluated using the aqueous two-phase partitioning method. Each showed a specific and characteristic pattern of surface properties. Correlations between the protein surface hydrophobicities and the coverages of the proteins by lipid coating with a synthetic detergent, dioleyl glucosyl L-glutamate, were discussed. The results indicated that the surface net hydrophobicity of each protein was indicative of the affinity of the protein for the coating detergent applied in lipid-coating. PMID- 10817818 TI - Comparison of biotin production by recombinant Sphingomonas sp. under various agitation conditions. AB - Biotin production by fermentation of recombinant Sphingomonas sp./pSP304 was investigated. A complex medium containing 60g/l of glycerol and 30g/l of yeast extract was suitable for biotin production. Biotin was produced in the late logarithmic or stationary phase after glycerol starvation. The optimum pH value for biotin production was 7.0. When the dissolved oxygen concentration (DO) was controlled at a constant level, the biotin concentration produced after 120h was significantly lower than that obtained in a test tube culture. Therefore, a batchwise jar-fermentor culture with a constant agitation speed and without DO control was conducted for investigating the effect of agitation conditions on biotin production. Six types of impeller were tested: turbine-blade type, turbo lift type, rotating mesh type (EGSTAR((R))), screw with draft tube type, Maxblend((R))type, and anchor type. With some impellers, agitation speed was also changed. Both the maximum cell concentration and biotin production varied depending on agitation conditions. Relatively high cell concentrations were attained with four of the impeller types, turbine-blade type, rotating mesh type, Maxblend((R)) type, and anchor type. Among these impellers, the turbine-blade impeller with sintered sparger was suitable for biotin production. After 120h, the cell concentration reached an OD(660) of 43 and a biotin concentration of 66mg/l was obtained, which was comparable with the results from the test tube culture. Morphological variation was also observed depending on the agitation conditions: oval-shaped, rod-shaped, and elongated-shaped cells. Biotin production was relatively high in slightly long rod-shape cells but low in elongated cells. The difference in morphology appeared to depend on the shear stress. It was found that biotin production was strongly correlated with cell length and the oxygen transfer coefficient (k(L)a); cell lengths in the range 4 7um and k(L)a values in the range 1.5-2.0/min were found to be suitable for biotin production in jar-fermentor culture. PMID- 10817819 TI - Kinetics of two-liquid-phase Taxus cuspidata cell culture for production of Taxol. AB - The effects of different organic solvents (paraffin, organic acid, alcohol and ester) and their volumetric fractions on the cell growth and Taxol production were studied in two-liquid-phase and the two-stage culture. A kinetic model, incorporated the effects of the toxicity of organic solvents was developed for two-liquid-phase culture of Taxus cuspidata in the two-stage Taxol production. The results showed that the proposed kinetic model could fit the experimental data satisfactorily. The results also showed that Taxol production could reach the optimal value when 10-logP was in the range of 2 to 5 and the volumetric fraction of the organic solvents at the corresponding the highest Taxol production should be lower when 10-logP was high. PMID- 10817820 TI - Efficient production of desulfurizing cells with the aid of expert system. AB - An expert system was used to achieve the high production of desulfurizing cells of Rhodococcus erythropolis KA 2-5-1. By adding a proper amount of sulfur containing component with the aid of the expert system, we could avoid excess feeding which resulted in the lowering of desulfurizing activity and starvation which caused serious damage to cell growth. In order to determine the addition amount by the expert system, the data of the amount of chemical elements contained in the cells were used as a reference for comparison with the medium components present. Culture experiments were carried out using a 5l jar fermentor with several kinds of media whose components were determined based on the inferred results with the aid of the expert system. We could restrict the amount of the sulfur component addition so that sulfur was a growth-limiting factor; in contrast, the amounts of other elements were sufficient for growth.As a result, the maximum specific production rate of 2-hydroxy biphenyl (2HBP) and the maximum cell concentration were 20mmolkg-drycells(-1)h(-1) and of 45g-drycellsl(-1), respectively. At 100h of cultivation, 1g/l of dibenzothiophene (DBT) was converted to 2HBP within 20h, i.e., 10mmolkg-drycells(-1)h(-1) of specific desulfurization activity, and the specific activity remained stable for a long period in the culture experiment. PMID- 10817821 TI - Releasing profiles of gene products from recombinant Escherichia coli in a high voltage pulsed electric field. AB - Release of recombinant proteins from gene-engineered Escherichia coli by applying a pulsed electric field (PEF) to a cell suspension was studied. When E. coli/pNC1, which produces beta-glucosidase and accumulates it in cytoplasm, was exposed to PEF, the most effective release of this enzyme was achieved in the cell suspension of 5% glycine and 15% PEG solution under 10kV/cm and 280J/ml of a PEF in a needle-plate electrode chamber. However, the amount of released beta glucosidase by PEF treatment was only 26% of that by ultrasonic treatment. On the other hand, alpha-amylase produced by E. coli/pHI301A and accumulated in the periplasmic space could be easily released by PEF treatment. When this recombinant E. coli was suspended in 0.9% NaCl and 10% PEG solution and exposed to 10kV/cm and 200J/ml of a PEF in a plate-plate electrode chamber, 89% of intracellular alpha-amylase with nine-times higher specific activity compared with that by ultrasonic treatment was released. The release tendency of cellobiohydrolase, produced by E. coli/pNB6 and accumulated in both the cytoplasm and periplasmic space, was intermediate between those of beta-glucosidase and alpha-amylase. In this case, 70% of cellobiohydrolase with 1.9-times higher specific activity compared with that by ultrasonic treatment could be released when E. coli/pNB6 was suspended in 15% PEG and 10kV/cm and 200J/ml of a PEF was applied in a needle-plate electrode chamber. These results indicated that PEF treatment could easily disrupt the outer membrane, but it was difficult to disrupt the cytoplasmic membrane simultaneously. Therefore, PEF treatment is useful for easy release of periplasmic protein with selectivity. PMID- 10817822 TI - Light intensity distribution in the externally illuminated cylindrical photo bioreactor and its application to hydrogen production by Rhodobacter capsulatus. AB - The light distribution in the externally illuminated cylindrical photo-bioreactor for production of hydrogen by a photosynthetic bacterium Rhodobacter capsulatus ST-410 was estimated. The estimation was performed on the basis of the Matsuura and Smith's diffuse model [1]. In the diffuse model, the incident light rays are assumed to proceed in every direction and the local intensity is calculated as the sum of the intensities of light. Since Lambert-Beer's law, extensively used in photometry, was not useful for explaining the decrease in the intensity of light by the biomass, an empirical expression was used. The measurement of the intensities from every direction was conducted in an externally illuminated cylindrical photo-bioreactor having an inner diameter of 60mm and a working volume of 550ml. The obtained results confirmed our estimation. The light distribution was applied to estimate the hydrogen production by R. capsulatus ST 410 using the same photo-bioreactor. The overall hydrogen-production rate was successfully estimated. PMID- 10817823 TI - Evaluation of kinetic parameters of biochemical reaction in three-phase fluidized bed biofilm reactor for wastewater treatment. AB - This study evaluates the kinetic parameters of biochemical reaction in three phase fluidized bed biofilm reactor from the steady state values of the response of the system to step changes in inlet concentration. It was observed from the outlet biological oxygen demand (BOD(5)) plot of the response of the system that as the inlet BOD(5) was increased, the outlet BOD(5) also increased, reached a peak value and then decreased until it leveled to a new steady state value corresponding to the new inlet concentration level. The increase in BOD(5) was attributed to the accumulation of substrate within the reactor as well as the decrease in biofilm substrate consumption rate as the microorganisms adjusted to the new environment. Using the substrate balance at steady state and assuming Monod kinetics, an equation relating the substrate consumption rate to substrate concentration (BOD(5)) and total biofilm surface area had been established. Monod kinetic parameters were found to be K=2.20g/m(2)/day, K(m)=17.41g/m(3) and K/K(m)=0.13m/day. The ratio K/K(m) can be taken as the indicator for biofilm substrate degradation effectiveness at low substrate concentrations. PMID- 10817824 TI - The ionic basis of cardiac activity in the bivalve mollusc Perna perna. AB - The ionic basis of cardiac activity and aspects of excitation-contraction (E-C) coupling were investigated in the isolated heart of the bivalve mollusc Perna perna, using the sucrose-gap technique. The role of the principal ions was established employing artificial seawater, in which specific ion concentrations were modified, and ion channel blockers. The mean membrane resting potential (MP) and the action potential (AP) were -33+/-0.7 mV (n=89) and 13+/-0.3 mV (n=71), respectively. The MP potential was primarily dependent on K(+) ions. Three types of cardiac APs were identified: fast, slow and spike-plateau potentials. Cardiac activity was maintained in Na(+)- or Ca(2+)-free salines but ceased when either Cd(2+) or EDTA was added to these salines. Other Ca(2+) channel blockers reduced the amplitude and increased duration of the cardiac APs. Tetrodotoxin (TTX) and procaine did not alter the AP. The data showed that the depolarizing phase of the AP was dependent on Ca(2+) influx while the plateau phase, when present, resulted from Na(+) influx that was modulated by Ca(2+). The mechanical responses were more sensitive to changes in extracellular Ca(2+) concentration than were the electrical responses. PMID- 10817825 TI - Exopolysaccharide production by the epipelic diatom Cylindrotheca closterium: effects of nutrient conditions. AB - During the stationary phase of a batch culture of the epipelic diatom Cylindrotheca closterium, accumulation of exopolysaccharides and intracellular carbohydrates was observed. When nitrogen was added to the culture in the stationary phase, growth was resumed and the accumulation of exopolysaccharides was delayed. This indicated that nitrogen depletion caused cessation of growth, and stimulated exopolysaccharide accumulation. Exopolysaccharide accumulation was also stimulated when cells were either resuspended in medium lacking N or P, or when they were inoculated in medium with low concentrations of N or P. Growth was not immediately affected by low N or P concentrations. S depletion only resulted in exopolysaccharide accumulation when growth was affected. Si or Fe depletion did not stimulate exopolysaccharide accumulation, even when growth rates were lowered. Apparently, stimulation of exopolysaccharide accumulation is dependent on the type of nutrient depletion. Intracellular storage carbohydrates did not accumulate when cells were incubated at low N or P concentrations. Cells grown with ammonium as nitrogen source produced more carbohydrates (both extracellular and intracellular) than cells grown with nitrate as nitrogen source, indicating that both exopolysaccharides and intracellular carbohydrates accumulated as a result of overflow metabolism. PMID- 10817826 TI - Separating the elements of habitat structure: independent effects of habitat complexity and structural components on rocky intertidal gastropods. AB - It has been difficult to understand the effects of habitat structure on assemblages because the different elements of habitat structure are often confounded. For example, few studies consider that the effects of structural components of a habitat (rocks, trees, pits, pneumatophores) may be separate from the complexity (e.g. surface area {SA}) they create. From prior observations and experiments, I developed three hypotheses about the effects of habitat structure on gastropods on rocky intertidal shores in Botany Bay, Australia. (1) The complexity of habitats positively affects the density and richness of gastropods. (2) The fractal dimension (D) represents elements of complexity that affect the density and richness of gastropods better than other indices of complexity. (3) The effects of specific structural components on the density and richness of gastropods are independent of their complexity. To test these hypotheses, treatments composed of pits and pneumatophores were used to independently manipulate complexity and structural components in experiments repeated at five different times on two shores. There was support for hypotheses (1) and (3) at most times and places but not for hypothesis (2). Richness, total density, and the densities of two of the three most common gastropods were greater in treatments with greater complexity. D was not definitively better than other indices of complexity, but D and SA were recommended for further consideration. When complexity was held constant, species richness and the density of most gastropods, except Austrocochlea porcata, was greater in treatments with pits than with pneumatophores. A common mechanistic explanation for the effects of habitat complexity on rocky intertidal gastropods relies on a specific characteristic of pits; they pool water and reduce desiccation stress. This assumption may be appropriate for many gastropods, but it was inappropriate for A. porcata. Habitat complexity affected its density, but this was not because of a characteristic specific to pits. The complexity and structural components of habitats have separate effects on assemblages, and it confuses the study of habitat structure to combine them. PMID- 10817827 TI - A long-term mesocosm study on the settlement and survival of juvenile European lobster Homarus gammarus L. in four natural substrata. AB - To date, the natural substratum preferences of early benthic phase (EBP) European lobsters (Homarus gammarus) remain largely unknown. This study utilised a large scale mesocosm experiment to determine if the animal favours cobble ground, similar to its American counterpart (Homarus americanus), or has other substratum preferences. Postlarvae were provided with the choice of settling on four natural substrata: sand, coralline algae, mussel shell and cobble. Over a nine month period, the number and size of juveniles on each substratum was recorded, with loss of chelipeds used as an indication of social interaction. After a 30 day period, a non-random distribution of lobsters was observed on the four substrata. Juveniles were more abundant in substrata which provided pre-existing shelter in the form of interstitial spaces, i.e. cobble and mussel shell, than in sand or coralline algae. The survival of individuals from postlarvae to 30 day old juveniles ranged from 5 to 14% with surviving benthic recruits showing a clear mode at 6-8 mm carapace length (CL) in size distribution. The density of lobsters per m(2) of cobble remained relatively constant (18/m(2)) throughout the study period while the density of juveniles on mussel shell decreased significantly (35 to 5/m(2)). The size distribution of lobsters on each substratum also varied with time. By the conclusion of the trial, lobsters found in mussel shell had a mode of 8-10 mm CL within a range of 6-14 mm CL while those in cobble had a mode of 10 12 mm CL within a range of 8-24 mm CL. Overall, the results underline the importance of shelter-providing habitat such as cobble or crevice-type substrata to EBP European lobsters. They also confirm that for a shelter-dwelling animal such as a lobster, the physical structure of the habitat is a key factor in determining both the size and number of its inhabitants. PMID- 10817828 TI - Filter-feeding and cruising swimming speeds of basking sharks compared with optimal models: they filter-feed slower than predicted for their size. AB - Movements of six basking sharks (4.0-6.5 m total body length, L(T)) swimming at the surface were tracked and horizontal velocities determined. Sharks were tracked for between 1.8 and 55 min with between 4 and 21 mean speed determinations per shark track. The mean filter-feeding swimming speed was 0.85 m s(-1) (+/-0.05 S.E., n=49 determinations) compared to the non-feeding (cruising) mean speed of 1.08 m s(-1) (+/-0.03 S.E., n=21 determinations). Both absolute (m s(-1)) and specific (L s(-1)) swimming speeds during filter-feeding were significantly lower than when cruise swimming with the mouth closed, indicating basking sharks select speeds approximately 24% lower when engaged in filter feeding. This reduction in speed during filter-feeding could be a behavioural response to avoid increased drag-induced energy costs associated with feeding at higher speeds. Non-feeding basking sharks (4 m L(T)) cruised at speeds close to, but slightly faster ( approximately 18%) than the optimum speed predicted by the Weihs (1977) [Weihs, D., 1977. Effects of size on the sustained swimming speeds of aquatic organisms. In: Pedley, T.J. (Ed.), Scale Effects in Animal Locomotion. Academic Press, London, pp. 333-338.] optimal cruising speed model. In contrast, filter-feeding basking sharks swam between 29 and 39% slower than the speed predicted by the Weihs and Webb (1983) [Weihs, D., Webb, P.W., 1983. Optimization of locomotion. In: Webb, P.W., Weihs, D. (Eds.), Fish Biomechanics. Praeger, New York, pp. 339-371.] optimal filter-feeding model. This significant under estimation in observed feeding speed compared to model predictions was most likely accounted for by surface drag effects reducing optimum speeds of tracked sharks, together with inaccurate parameter estimates used in the general model to predict optimal speeds of basking sharks from body size extrapolations. PMID- 10817829 TI - Pigment signatures associated with an anoxic coastal zone: Bahia Concepcion, Gulf of California. AB - Bahia Concepcion is a coastal lagoon that has bottom anoxic conditions and high pigment concentrations during the summer. The phytoplankton responsible for this pigment increase is enigmatic, therefore we sampled the lagoon to analyze the pigment with a C8-HPLC system to look for signatures of phytoplankton groups. Analysis reveals a low pigment concentration in the mixed layer with a higher concentration of zeaxanthin and increasing values of chlorophyll a, peridinin, and fucoxanthin below, which peaked at the depth where oxygen dramatically decreases and H(2)S increases. Below this depth, a high pigmentation was recorded and the most important signatures were six chlorophyll-like pigments that eluted between the fucoxanthin and the chlorophyll a, and one carotenoid that eluted just after the chlorophyll a. Spectral characteristics of these last pigments are very similar to pigments present in the Chlorobiales group. These results suggest that cyanobacteria, diatoms, and dinoflagellates are responsible for the chlorophyll a increases, though in highly pigmented samples, anoxygenic phototrophic bacteria are probably the main contributors to the increase in pigments. PMID- 10817830 TI - Settlement preferences and early migration of the tropical sea cucumber Holothuria scabra. AB - Settlement and post-settlement processes of the sea cucumber Holothuria scabra Jaeger were studied in the laboratory. Independent and paired choice experiments revealed that several substrates could induce metamorphosis into pentactulae, but that specific substrates favoured settlement. Leaves of seagrass Thalassia hemprichii, with or without their natural bio-film, yielded the highest settlement rates (4.8-10.5%). T. hemprichii was preferred as a settlement substrate over sand, crushed coral, several other plant species and artificial seagrass leaves with or without a bio-film. Only settlement on the seagrass, Enhalus acoroides, was similar to that recorded for T. hemprichii. In the absence of a substrate, the larvae delayed settlement for nearly 96 h and survival was less than 0.5%. Sand and crushed coral, either alone or together, induced settlement from <1.5% of the available larvae. The pentactulae found on sand, coral and in bare containers were 10-35% smaller than those on T. hemprichii leaves. Soluble extracts from T. hemprichii and E. acoroides successfully induced metamorphosis and settlement on clean plastic surfaces. Newly settled juveniles remained on the seagrass leaves for 4-5 weeks before migrating to sand at around 6 mm in length. Prior to this, the juveniles spent 4-5 days moving on and off the leaves. Once on the sand, the juveniles became deposit-feeders, but did not show the typical burrowing behaviour of older specimens until they reached around 11 mm in length. The larvae of H. scabra appear to actively select seagrass leaves, possibly through chemical detection. We hypothesise that larvae settling on seagrass have an increased chance of growth and survival because they are provided with a suitable substrate on which to grow, and a bridge to sand substrates as they become deposit-feeders. PMID- 10817831 TI - Experimental examination of the effects of atmospheric wet deposition on primary production in the Yellow Sea. AB - The effects of atmospheric wet deposition on primary production in the coastal Yellow Sea were examined by in situ incubation experiments in August, 1997. Phytoplankton species flourished in response to nutrient additions and chlorophyll-a (Chl-a) increased significantly when rainwater was added. Concentration of Chl-a increased 2.6 times with the addition of 10% (v/v) rainwater. In a coastal eutrophic region like Jiaozhou Bay, the impact of atmospheric wet deposition is negligible. However, the N/P ratio ranged from 22:1 to 80:1, indicating P limitation of photosynthesis in the Yellow Sea. During field observations, high N and low P rainwater was observed to be transported into the oligotrophic central Yellow Sea. Ammonium was used by phytoplankton in preference to nitrate, and Fe stimulated nutrient uptake by phytoplankton. Rainwater increased the Chl-a growth more than a single nutrient owing to the abundance of both plant and trace nutrients in the rainwater. A negative exponent relationship may exist between the impact of rainwater and total nutrient quality of the water in influencing phytoplankton growth. PMID- 10817832 TI - Effects of macroalgal mats on intertidal sandflats: an experimental study. AB - The growth of green macroalgal mats is becoming increasingly common in many marine intertidal habitats. While the ecological effects of such growth has previously been experimentally investigated on mudflats, such experiments have rarely been performed on intertidal sandflats. This study investigated the ecological effects of macroalgal cover on a moderately exposed intertidal sandflat, Drum Sands, Firth of Forth, Scotland. Artificially implanted Enteromorpha prolifera (Muller) caused marked changes in the macrobenthos, together with significant changes in all the measured sediment variables. After 6 weeks, the weed significantly increased the macrofaunal diversity. The numbers of Pygospio elegans (Claparede) were significantly reduced under weed mats, while those of Capitella capitata (Fabricius), oligochaetes and gammarids increased. Percent water, organics and silt/clay contents, medium phi and sorting coefficient significantly increased in the sediments under weed mats which also became significantly more reduced between 1 and 8 cm depth. After 20 weeks, a macrofaunal community numerically dominated by C. capitata, with a significantly reduced diversity, was present under weed mats, while sediment variables were no longer significantly different from controls. The negative effect of E. prolifera on P. elegans was mainly due to larval filtering, suggesting that weed is likely to have detrimental effects on population maintenance of most species which rely on planktonic larval recruitment. These results are broadly similar to those obtained from algal manipulation experiments performed in much more sheltered, muddier environments. We suggest that a predictable deterioration in environmental quality results from the growth of macroalgal mats in soft-bottom habitats. However, the longer term effects of such algal growth are less predictable and depend upon the spatial distributions of the most abundant infaunal species and the spatial heterogeneity of weed mat establishment. PMID- 10817833 TI - Salivary glands in ixodid ticks: control and mechanism of secretion. AB - The salivary glands are vital to the biological success of ixodid ticks and the major route for pathogen transmission. Important functions include the absorption of water vapor from unsaturated air by free-living ticks, excretion of excess fluid for blood meal concentration, and the secretion of bioactive protein and lipid compounds during tick feeding. Fluid secretion is controlled by nerves. Dopamine is the neurotransmitter at the neuroeffector junction regulating secretion via adenylate cyclase and an increase in cellular cAMP. Dopamine also affects the release of arachidonic acid which is subsequently converted to prostaglandins. Prostaglandin E(2) (PGE(2)) is secreted at extremely high levels into tick saliva for export to the host where it impacts the host physiology. Additionally, PGE(2) has an autocrine or paracrine role within the salivary gland itself where it interacts with a PGE(2) receptor to induce secretion (exocytosis) of bioactive saliva proteins via a phosphoinositide signalling pathway and an increase in cellular Ca(2+). Regulation of fluid secretion has been extensively studied, but little is known about the mechanism of fluid secretion. Continuing advances in tick salivary gland physiology will be made as key regulatory and secretory gland proteins are purified and/or their genes cloned and sequenced. PMID- 10817834 TI - Polyamines, and effects from reducing their synthesis during egg development in the yellow fever mosquito, Aedes aegypti. AB - Development of eggs after a blood meal in the yellow fever mosquito Aedes aegypti involves hormonal changes, synthesis of nucleic acids, activation of the digestive enzyme trypsin, and production of the yolk protein vitellogenin. Polyamines have been implicated in growth processes and were here examined for possible involvement during egg development. The data suggest that polyamines are important for normal vitellogenesis in the mosquito. Polyamine levels and activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase, key enzymes in the polyamine pathway, were determined in the fat body for two days after a blood meal. During the time that the macromolecules required for vitellogenesis were being synthesized, polyamine levels increased as did the activities of their rate-limiting enzymes. Administration of suicide inhibitors of ornithine decarboxylase, alpha-difluoromethylornithine (DFMO) and alpha monofluoromethyldehydroornithine methylester (MDME), limited increased polyamine levels and disrupted macromolecular syntheses, particularly during the first twenty-four hours after blood feeding. Specific metabolic processes reduced by DFMO included trypsin activity, and production of RNA, DNA and vitellogenin. MDME had differential effects on transcription of some mRNA species made after an oogenic meal. The level of actin mRNA was not affected by inhibiting polyamine synthesis, but the mRNA levels of vitellogenin, trypsin, and the vitelline membrane protein were decreased. Adding polyamines to a meal containing DFMO or MDME partially reversed the effects of these inhibitors. Increases in spermidine and spermine were associated with these reversals. PMID- 10817835 TI - Hydrocarbon site of synthesis and circulation in the desert ant Cataglyphis niger. AB - Chemical analyses revealed that in Cataglyphis niger both the hemolymph and the crop contain the same hydrocarbons that are found in the postpharyngeal gland (PPG). On the cuticle, on the other hand, alkanes, and in particular nonacosane, were more abundant than in the PPG. Studies of their biosynthesis in vivo, using intact ants, revealed the presence of newly synthesized hydrocarbons in both the PPG and the crop. In decapitated ants (in the absence of the PPG), however, the crop did not contain any newly synthesized hydrocarbons, indicating the PPG as the major source of crop hydrocarbons. The fat body, as demonstrated by in vitro studies, is the major tissue that biosynthesizes hydrocarbons. The PPG failed to do so, but showed good de novo biosynthesis of other lipid constituents. The large amount of hydrocarbons in the crop suggests that the alimentary canal may serve as an outlet for the overflow of PPG hydrocarbons, or as a route for the directed clearance of hydrocarbons from the PPG.These results confirm and enlarge the model proposed for hydrocarbon circulation in C. niger. They are synthesized by the fat body, released to the hemolymph and transported to the cuticle and the PPG. The PPG hydrocarbons are applied to the cuticle by self-grooming, but can also be cleared via the alimentary canal. Partial emptying of the PPG may facilitate the admixing of recognition cues that the ant may acquire from nestmates by trophallaxis. The reason for the dissimilarity in hydrocarbon composition between the PPG and the cuticle is not yet clear; it may be due to secretions from additional glands, or reflect deviant hydrocarbon transport mechanisms between the PPG and the cuticle. PMID- 10817836 TI - Foraging in the ant Camponotus mus: nectar-intake rate and crop filling depend on colony starvation. AB - The effects of colony starvation on the dynamics of nectar collection were studied in individual workers of the ant Camponotus mus. A laboratory colony was first deprived of carbohydrates for 15days, and thereafter fed daily ad libitum with diluted honey until satiation. During these two successive experimental phases, the probability of feeding, crop filling and fluid-intake rates were recorded daily for individual foragers collecting a 10% (w/w) sucrose solution. The feeding responses of individuals varied with the nutritional state of the colony. When the colony was deprived of sugar, acceptance of the sucrose solution was higher than under satiation. Feeding time increased with increasing starvation. During deprivation workers fed nearly continuously on the solution, whereas a number of feeding interruptions occurred under satiation. Crop filling also increased with increasing starvation, and showed a marked decrease when the colony was satiated. Fluid-intake rate during the deprivation phase was roughly twice that during the satiation phase. This matched well with the difference in sucking frequency recorded during ingestion in satiated and starved workers, which was also higher during starvation. Results indicate that the responsiveness of foragers, determined by the nutritional state of the colony, influenced both foraging decisions and the dynamics of fluid intake. PMID- 10817837 TI - Expression of Drosophila homologue of senescence marker protein-30 during cold acclimation. AB - Gene expression during cold acclimation at a moderately low temperature (15 degrees C) was studied in Drosophila melanogaster using a subtraction technique. A gene homologous to senescence marker protein-30 (SMP30), which has a Ca(2+) binding function, was up-regulated at the transcription level after acclimation to 15 degrees C. This gene (henceforth referred to as Dca) was also expressed at a higher level in individuals reared at 15 degrees C from the egg stage than in those reared at 25 degrees C. Moreover, DCA mRNA increased at the senescent stage in Drosophila, although SMP30 is reported to decrease at senescent stages in mammals. In situ hybridization to polytene chromosomes revealed that the Dca gene was located at 88D on chromosome 3R. The 5' flanking region of this gene had AP-1 (a transcription factor of SMP30) binding sites, stress response element and some other transcription factor binding sites. The function of DCA was discussed in relation to the possible regulation of cytosolic Ca(2+) concentration. PMID- 10817838 TI - Comparison of feeding behaviour of Triatoma infestans, Triatoma brasiliensis and Triatoma pseudomaculata in different hosts by electronic monitoring of the cibarial pump. AB - Feeding behaviour of Triatoma infestans, T. brasiliensis and T. pseudomaculata on pigeons and mice was compared by electronic monitoring of the cibarial pump. The methodology developed permits the study detailed of triatomine feeding behaviour using an artificial feeder as well as on live hosts. T. infestans was the species that fed most rapidly on the two hosts tested (28.03+/-1.6 mg/min for pigeon and 21.33+/-1.7 mg/min for mouse), followed by T. brasiliensis (17.09+/-1.4 mg/min and 13.1+/-1.5 mg/min for pigeon and mouse, respectively) and T. pseudomaculata, (5.23+/-0.6 mg/min and 4.09+/-0.4 mg/min for pigeon and mouse, respectively). The quantity of liquid ingested per stroke of the cibarial pump was 100.7+/-4.1 nl for T. infestans, 69.1+/-2.7 nl for T. brasiliensis and 26.8+/-1.5 nl for T. pseudomaculata. The rate of engorgement in pigeons tended to be greater than that obtained for mice in the three species studied. In the experiments carried out using mice, probing times tended to be longer and interruptions during the meal more prolonged. This reinforces the idea that feeding on mice is more difficult than on pigeons, requiring more contact time to obtain the similar quantity of blood. PMID- 10817839 TI - Involvement of both granular cells and plasmatocytes in phagocytic reactions in the greater wax moth, Galleria mellonella. AB - Although it has been previously found by most authors that only plasmatocytes are involved in phagocytosis of non-self in the greater wax moth, Galleria mellonella, in the present study we demonstrate that in vitro, both granular cells and plasmatocytes are involved in this reaction, using monolayers of these haemocytes prepared from larval haemolymph by a differential cell fractionation method. The adhesion of granular cells to glassware and phagocytosis by granular cells of FITC-labelled silica beads were both greatly reduced by the presence of p-NPGB, a serine proteinase inhibitor, which is known to inhibit the activation of the prophenoloxidase cascade, but the reactions were only partly influenced by PTU, an inhibitor of phenoloxidase. These results suggest that an enhancing factor for both reactions is phenoloxidase itself or a component induced during the course of activation of the prophenoloxidase cascade, but not the melanised substance produced by the action of this reaction. For plasmatocytes, attachment to non-self was totally blocked by the absence of CaCl(2) or by the presence of EDTA at concentrations greater than 20 mM, and phagocytosis was greatly enhanced by CaCl(2), but suppressed by EDTA. These results suggest that calcium is a factor required for adhesion of plasmatocytes, and that it also functions to enhance their phagocytic action. PMID- 10817840 TI - Spectral correspondence between visual spectral sensitivity and bioluminescence emission spectra in the click beetle Pyrophorus punctatissimus (Coleoptera: Elateridae). AB - The presence of two spectral mechanisms, near-ultraviolet and green (lambda(max)=545nm), is strongly suggested by electroretinographic visual spectral sensitivity curves obtained under dark and red chromatic adaptation conditions in the compound eyes of the click beetle Pyrophorus punctatissimus. The bioluminescence emission of the dorsal prothoracic lanterns is deep green (lambda(max)=543nm) and that of the ventral abdominal lantern is lime green (lambda(max)=556nm) in colour in P. punctatissimus. A broad green visual receptor would detect both deep green and lime green bioluminescent optical signals. PMID- 10817841 TI - Time-course haemolymph juvenile hormone titres in solitarious and gregarious adults of Schistocerca gregaria, and their relation to pheromone emission, CA volumetric changes and oocyte growth. AB - The haemolymph JH III titres in solitarious and gregarious adult desert locusts, Schistocerca gregaria, were examined in relation to corpus allatum (CA) volumes, aggregation-maturation pheromone production in males and oocyte growth in females. The JH titres of gregarious females were generally higher than those of solitarious females at all ages studied. The titre patterns, however, were similar: relatively high on day 10, dropping to low levels between days 20 and 25, before rising again by day 25. In the solitarious males, the JH titre was very low on day 10 after fledging, but increased gradually and reached a maximal amount on day 30. The JH titre in gregarious males was low on day 10, elevated on day 15 coinciding with the start of the production of the pheromone, and dropped to a relatively low level on day 20 around the time of maximal pheromone production, then rising again by day 25. These results suggest that biosynthesis of the pheromone is associated with a high JH titre peak in the haemolymph. Although a clear relationship was found during the first gonadotropic cycle between JH titres, on one hand, and CA volume and oocyte growth, on the other, in both phases, no such correlation could be discerned in the second cycle. PMID- 10817842 TI - Linking gustatory neurobiology to behavior in vertebrates. AB - Technological advances in neuroscience in general, and molecular biology in particular, offer tremendous experimental opportunities for researchers studying the vertebrate gustatory system. Ultimately, however, the neurobiological events must be linked to the taste-related behavior of the animal. Although there has been some promising work in this regard, progress has been hampered by an absence of a unified theoretical framework regarding function, unconfirmed assumptions inherent in many experimental designs, and a misguided predilection for researchers to interpret results from a variety of vertebrate models in the context of human psychophysics. This review article offers a heuristic for the organization of taste function and encourages greater coordination between behavioral and neurobiological approaches to the problem of understanding gustatory processes in the nervous system. The potential power of such coordinated efforts is discussed as well as the possible interpretive pitfalls associated with the neural analysis of gustation. PMID- 10817843 TI - The adolescent brain and age-related behavioral manifestations. AB - To successfully negotiate the developmental transition between youth and adulthood, adolescents must maneuver this often stressful period while acquiring skills necessary for independence. Certain behavioral features, including age related increases in social behavior and risk-taking/novelty-seeking, are common among adolescents of diverse mammalian species and may aid in this process. Reduced positive incentive values from stimuli may lead adolescents to pursue new appetitive reinforcers through drug use and other risk-taking behaviors, with their relative insensitivity to drugs supporting comparatively greater per occasion use. Pubertal increases in gonadal hormones are a hallmark of adolescence, although there is little evidence for a simple association of these hormones with behavioral change during adolescence. Prominent developmental transformations are seen in prefrontal cortex and limbic brain regions of adolescents across a variety of species, alterations that include an apparent shift in the balance between mesocortical and mesolimbic dopamine systems. Developmental changes in these stressor-sensitive regions, which are critical for attributing incentive salience to drugs and other stimuli, likely contribute to the unique characteristics of adolescence. PMID- 10817844 TI - Is the avian hippocampus a functional homologue of the mammalian hippocampus? AB - The effects of hippocampal lesions on the processing and retention of visual and spatial information in birds and mammals is reviewed. Both birds and mammals with damage to the hippocampus are severely impaired on a variety of spatial tasks, such as navigation, maze learning, and the retention of spatial information. In contrast, both birds and mammals with damage to the hippocampus are not impaired on a variety of visual tasks, such as delayed matching-to-sample, concurrent discrimination, or retention of a visual discrimination. In addition, both birds and mammals with hippocampal damage display impairments in the acquisition of an autoshaped response, as well as alterations in response suppression. These findings suggest that the avian hippocampus is a functional homologue of the mammalian hippocampus, and that in both birds and mammals the hippocampus is important for the processing and retention of spatial, rather than purely visual information. PMID- 10817845 TI - Sex-related hormonal influences on pain and analgesic responses. AB - Considerable evidence indicates sex-related differences in pain responses and in the effectiveness of various analgesic agents. Specifically, females are at greater risk for experiencing many forms of clinical pain and are more sensitive to experimentally induced pain relative to males. Regarding analgesic responses, nonhuman animal studies indicate greater opioid analgesia for males, while a limited human literature suggests the opposite. Though the mechanisms underlying these effects remain unclear, the influence of gonadal hormones on nociceptive processing represents one plausible pathway whereby such sex differences could emerge. The present article reviews the complex literature concerning sex steroid effects on pain responses and analgesia. First, nonhuman animal research related to hormonal effects on nociceptive sensitivity and analgesic responses is presented. Next, human studies regarding gonadal hormonal influences on experimental pain responses are reviewed. Several potential mechanisms underlying hormonal effects on nociceptive processing are discussed, including hormonal effects to both peripheral and central nervous system pathways involved in pain transmission. Finally, based on these findings we draw several conclusions and make specific recommendations that will guide future research as it attempts to elucidate the magnitude and importance of sex-related hormonal effects on the experience of pain. PMID- 10817846 TI - Beta-blockers - again, a lesson to us all, especially the research funding community. PMID- 10817847 TI - Non-invasive assessment of myocardial blood flow using echocardiography. PMID- 10817848 TI - Reduction of the infarcted area with the use of simplified coronary sinus retroperfusion during experimental coronary artery occlusion. AB - This study examined if the use of simplified coronary sinus retroperfusion would lead to any reduction in the infarcted area associated with improved right and left ventricular function. Twelve mongrel dogs were entered in this study. Following anesthesia, a fast response thermistor was placed on the pulmonary artery via the jugular vein and aorta via the left ventricular apex. The left anterior descending artery (LAD) was separated from the vein. A retrograde cardioplegia catheter was inserted into the coronary sinus. Following these procedures, LAD was occluded for a period of 3.5 h. After 30 min ischemia, the aorta-coronary sinus connection was established. The animals were divided into two equal groups. One group was not treated and was considered the control group (six animals). In the remaining group (six animals), retroperfusion was used and was considered the retroperfusion group. At the end of the study, the left ventricular ejection fraction was 65+/-15% in the retroperfusion group and 48+/ 5% in the control group (P<0.05). The left ventricular stroke work index was 0.44+/-0.04 (g m/kg) in the retroperfusion group and 0.31+/-0.05 (g m/kg) in the control group (P<0.05). Cardiac output was 1650+/-75 ml/min in the retroperfusion group and 1250+/-125 ml/min in the control group. The ratio of the infarct size to the area at risk was 49+/-5% in the control group and 7+/-3% in the retroperfusion group. In light of these studies, we conclude that simplified coronary sinus retroperfusion appears to be an effective method that must be taken into consideration. PMID- 10817850 TI - Nitric oxide in heart failure PMID- 10817849 TI - Endogenous nitric oxide in patients with chronic heart failure (CHF): relation to functional impairment and nitrate-containing therapies. AB - We assessed the levels of exhaled nitric oxide (eNO) in patients with chronic heart failure (CHF) according to the functional impairment and the use of nitrate containing agents. Forty patients (age 55+/-9 years) were classified according to the NYHA classes I-II (n=18, group 1) and classes III-IV (n=22, group 2), and to the use of nitrate-containing drugs (Nitrate+, Nitrate-). Twenty-two healthy age related subjects served as controls (group 3). Respiratory function, symptom limited incremental cycloergometry and resting eNO concentration at peak (FENOp) or plateau (FENOpl) of the single-breath exhalation curve were assessed in all subjects. FENOpl was significantly lower in patients than in controls (7.8+/-2.7 and 10.6+/-2.8 ppb, respectively, P<0.005) and lower in most severe CHF patients (7.1+/-2.6 and 8.8+/-2.7 ppb in group 2 and group 1, respectively, P<0.05). A significant correlation between peak V'O(2), Watts and FENOpl (r=0.42, P<0.013 and r=0.46, P=0.008, respectively) was found. Independent of NYHA class, Nitrate+ showed higher FENOp levels than Nitrate- patients (36.9+/-15.7 vs. 28. 1+/-15.1 ppb, P<0.05). Resting eNO was lower in the most compromised CHF patients and was significantly related to exercise capacity. Nitrate-containing agents might influence the levels of eNO in these patients. PMID- 10817851 TI - Left atrial appendage function in patients with different pacing modes. AB - Many studies suggest that patients who receive a ventricular pacemaker have a higher incidence of systemic thromboembolism compared to patients receiving a physiological pacemaker. However, the exact mechanism regarding the etiology of thromboembolism remains unclear. We evaluated the left atrial appendage (LAA) functions, using multiplane transesophageal echocardiography (TEE), in patients with different pacing modes. In order to evaluate the ejection fraction (EF), peak emptying (V(E)) and filling (V(F)) flow velocities of the LAA by TEE, we studied 31 patients (mean age 63+/-18.5 years) who had been paced for 5.0+/-2.9 years. Patients with atrial fibrillation, left ventricular dysfunction and mitral valve disease were excluded. The pacing indications were complete atrioventricular block (AVB) in 19 patients (9 VVI, 10 VDD or DDD) and sick sinus syndrome (SSS) in 12 patients (5 VVI, 7 DDD). Mean EF, V(E) and V(F) of the LAA were significantly lower in all patients with ventricular pacing (25.5+/-15.6%, 30.4+/-15.6 cm/s and 29. 1+/-19.2 cm/s, respectively) compared to those with physiologic pacing (48.5+/-16.9%, 59.6+/-16.3 cm/s, 57.9+/-18.5 cm/s, respectively) (P<0.01 in all). When patients were further classified with respect to underlying heart disease whether they had SSS or AVB, all measurements of the LAA (EF, V(E) and V(F)) in both subgroup of patients with SSS and AVB were found significantly lower in those with ventricular pacing than in those with physiologic pacing (Tables 3 and 4). This decrease, especially in LAA flow, was much greater in those with SSS (Mean V(E) and V(F) <20 cm/s). In a patient paced with VVI for SSS, a thrombus was detected within the LAA cavity. In conclusion, these results suggest that the pacing modality appeared to influence the LAA functions in paced patients. Patients with asynchronous ventricular pacing modes had a significantly higher incidence of depressed LAA functions than did patients with physiological pacing, especially more marked in patients with sick sinus syndrome. This may be a factor responsible for increased risk of thrombus formation and thromboembolic events in this patient population. PMID- 10817852 TI - Evaluation of diastolic dysfunction and repolarization dispersion in Behcet's disease. AB - Behcet's disease is a generalized chronic inflammatory disease characterized by genital, ocular, and cardiovascular involvement. Recently, left ventricular diastolic dysfunction, ventricular arrhythmia and sudden cardiac death have been documented in Behcet's disease. From January 1996 to May 1998, we investigated left ventricular systolic and diastolic function, valvular heart disease, ischemic heart disease and repolarization dispersion in 71 cases, 40 men and 31 women (mean age, 36.8+/-10.3 years) with Behcet's disease. All of the results were compared with the control group of 33 men and 22 women (mean age, 37.9+/-9.6 years). Exercise stress test or myocardial perfusion scintigraphy was performed for the documentation of ischemia. All the patients and the controls were recorded by M-mode, 2-D and Doppler echocardiography. Ventricular wall thickness, valvular apparatus, left ventricular systolic and diastolic parameters were evaluated. Repolarization dispersion parameters were calculated as the difference between maximal and minimal values of QT from 12-lead electrocardiogram recording at baseline, immediate and end of recovery from the exercise stress tests. The measured parameters were compared with the control group by using statistical methods. In the Behcet's group of 22 patients (31%) E/A ratio was <1. In the control group of five cases (10%) E/A ratio was <1 (P=0.003). In the Behcet's group isovolumic relaxation time (IRT) and mitral deceleration time (MDT) were longer than the control group (P=0.002, P=0.041, respectively). A mean QT of 368+/-30 ms and mean QT dispersion of 73+/-14 ms in the patient group compared with a mean QT of 395+/-39 ms and mean QT dispersion of 38+/-12 ms in the controls. There was no statistical difference between the mean QT values of the patient and control groups however, ventricular dispersion parameters in the Behcet's patients were longer than in the controls (P<0.001). There was also statistical significance for the QT dispersion between the Behcet's patients with and without diastolic dysfunction (P<0.01). In conclusion, the study reveals that the patients with Behcet's disease have a high incidence of increased diastolic dysfunction and repolarization dispersion. A positive correlation may exist between diastolic dysfunction and QT dispersion. PMID- 10817853 TI - Exercise-induced negative U waves in precordial leads as a marker of viable myocardium in patients with recent anterior myocardial infarction. AB - Patients with recent anterior myocardial infarction and a significant stenosis in the left anterior descending coronary artery were divided into two groups according to the presence (Group A, n=24) or absence (Group B, n=77) of negative U waves in the precordial leads during exercise stress test to compare the coronary arteriographic findings. Both total occlusion of the left anterior descending coronary artery (79% vs. 31%, P<0.01)) and good (index=2, 3) collateral circulation to the territory of this artery (92% vs. 36%, P<0.01) were observed more often in group A than in group B. In order to determine whether detection of exercise-induced negative U waves in precordial leads can predict the presence of viable myocardium, the 56 patients (20 of group A and 36 of group B) with total or subtotal occlusion of the left anterior descending coronary artery were studied further. The coronary arteriographic and exercise 201Tl scintigraphic findings were compared between the groups. Good collateral circulation to the territory of this artery was observed significantly (P<0.05) more often in group A (100%) than in group B (61%). Patients with multivessel disease were significantly (P<0.05) more prevalent in group A (70%) than in group B (33%). The incidence of a significant partial redistribution in the anteroseptal area in the 20Tl images 4 h after exercise was significantly (P<0.01) higher in group A (85%) than in group B (39%). In 29 patients with anterior Q-wave myocardial infarction and exercise-induced ST elevation in precordial leads, a significant 201Tl redistribution in the anteroseptal area was observed in 8 (80%) of 10 patients of group A in contrast to only 4 (21%, P<0.05) of 19 in group B. In the diagnosis of the viability associated with 201Tl redistribution in the anteroseptal area by the finding of exercise-induced negative U waves, the sensitivity was 67% and the specificity was 88% in these patients. We conclude that exercise-induced negative U waves in precordial leads are a convenient and specific marker for the presence of viable myocardium in patients with recent anterior myocardial infarction. PMID- 10817854 TI - Sympathetic inhibition with clonidine prolongs survival in experimental chronic heart failure. AB - Activation of the sympathetic nervous system is associated with increased mortality in congestive heart failure (CHF), and inhibition of the sympathetic nervous system by centrally acting sympatholytic agents has been shown to have beneficial effects on hemodynamics in these patients. However, the effect of sympathetic inhibition on survival in CHF is not clear. In the present study, the effect of sympathetic inhibition with clonidine on survival was examined in a rat model of heart failure. Myocardial infarction and heart failure was induced in rats by ligation of the left coronary artery and sham-operated rats served as the control. Two weeks after surgery, the ligated rats were randomly assigned to the clonidine (100 microg kg(-1) d(-1), n=30) group or the placebo (vehicle, n=31) group. All rats were followed daily for a 1-year period or until spontaneous death. Compared with placebo therapy, clonidine treatment reduced systolic blood pressure and heart rate throughout the experimental period. The plasma norepinephrine level determined at the end of the experiment was also reduced. Long-term sympathetic inhibition with clonidine treatment improved 1-year survival (50% vs. 22.6%, P<0.05) after surgery in this rat model of CHF. PMID- 10817855 TI - Sympathetic activation in chronic heart failure PMID- 10817856 TI - Prior cytomegalovirus, Chlamydia pneumoniae or Helicobacter pylori infection and the risk of restenosis after percutaneous transluminal coronary angioplasty. AB - We investigated a possible correlation between the serologic status concerning Cytomegalovirus (CMV), Chlamydia pneumoniae (CP) and Helicobacter pylori (HP) and the occurrence of restenosis in patients undergoing percutaneous transluminal coronary angioplasty for symptomatic coronary artery disease. Tests for anti-CMV IgG, anti-Chlamydia pneumoniae IgG and IgA and HP IgG and IgA were performed with an enzyme-linked immunosorbent assay (ELISA). Restenosis was defined as >/=50% stenosis at follow-up angiography in a vessel with less than 50% stenosis immediately after PTCA. Of 148 patients, 112 (75.7%) were seropositive for CMV, 75 (50.7%) were seropositive for CP and 78 (52.7%) were seropositive for HP. Restenosis occured in 31.8% of patients. CMV seropositivity was established in 74.5% of patients with restenosis versus 76.2% without restenosis (P=0.82), CP seropositivity was established in 44. 7% of patients with restenosis versus 53.5% without restenosis (P=0. 32), HP seropositivity was established in 53.2% of patients with restenosis versus 52.5% without restenosis (P=0.94). In contrast to some earlier studies CMV or HP seropositivity could not be found to be associated with the risk of restenosis after coronary intervention. An association between the serological status of CP and restenosis could also not be established. PMID- 10817857 TI - Renal function after cardiopulmonary bypass surgery in cyanotic congenital heart disease. AB - We studied perioperative renal damage in 22 patients with long-standing cyanotic congenital heart disease. BACKGROUND: [corrected] Postoperative acute renal failure is a major complication of cardiac surgery associated with poor prognosis. Our study was designed to identify risk factors for renal failure in patients with cyanotic congenital heart disease. PATIENTS: 22 cyanotic patients with a oxygen saturation of 82% (38-92%), age 14 years (5-42 years) and six controls with atrial septal defect, age 37 years (28-66 years) were investigated with repeated urinary analyses. RESULTS: Before operation, six of 22 of the cyanotic patients had albuminuria. Postoperatively three patients developed acute renal failure including glomerular and tubular damage. Urinary albumin analysis >1000 mg g(-1) creatinine after 24 h and N-acteyl-beta-D-glucosaminidase analysis >100 U g(-1) creatinine after 48 h predicted dialysis requirement. In noncyanotic controls only one preoperative and none of the postoperative analyses were pathological. CONCLUSIONS: Patients with cyanotic congenital heart disease are at risk for acute renal failure, which can be prognosed by urine analysis already 24 h after surgery. Cyanotic glomerulopathy should be known before surgery. To lower the risk, cardiopulmonary bypass time should be kept as short as possible and adequate hydration should be maintained in combination with extended diuretic therapy. PMID- 10817858 TI - Concomitant management of airway and its adjacent vascular pathology in addition to repair of congenital cardiac defects. AB - Respiratory symptoms are often present in infants with congenital cardiac anomalies, but the intrinsic pathology of the airway itself or external compression by abnormal vessels is frequently undetected before cardiac repair. We collected 12 patients with airway pathology from July 1996 to October 1998; all had definite diagnosis of lesions of the airway and its adjacent vessels by preoperative ultrafast computed tomography. Four had intrinsic pathology (one retrotracheal diverticulum, three tracheal stenosis) as well as external compression. Among them, six had complete vascular ring, four partial rings and three had bronchial compression by aneurysmal dilatation of branch pulmonary arteries. Simultaneous airway repair (one diverticulectomy, three patch tracheoplasty) and external decompression were performed in ten cases under cardiopulmonary bypass, nine of ten had simultaneous cardiac repair; the other two were done before cardiac repair. All survived except three. We concluded that ultrafast computed tomography is indispensable for definite diagnosis of airway and adjacent cardiovascular pathology. Mere repair of congenital cardiac defects without rectification of the airway and its adjacent structures is incompatible with survival. PMID- 10817859 TI - G protein beta3 subunit variant and essential hypertension in Taiwan - a case control study. AB - Recent studies have shown that a C825T polymorphism of the gene encoding the G protein beta3 subunit contributes to the genesis of essential hypertension. However, the link between the gene and blood pressure is not consistently found in different populations. The aim of the present study is to investigate this issue in Taiwan. We analyzed the allelic status in 302 hypertensive (age, 60+/-11 years; male/female, 136/166) and 199 normotensive subjects (62+/-15 years; male/female, 90/109). Our result showed that the T allelic was more frequently seen in the hypertensive group than the normotensive, but the difference did not reach statistic significance (56.5 vs. 54.3%, P>0.1). Subsequent analysis demonstrated a similar trend in the female (58.7 vs. 53.7%, P>0.1) but a reverse trend in the male (53.7 vs. 55%, P>0.1). Another finding was that the T allele frequency in all the groups was over 50%, markedly higher than those reported in whites. In conclusion, the observation suggests that the polymorphism in the G protein gene is not likely to play an important role in the manifestation of high blood pressure in Taiwan. PMID- 10817860 TI - The G protein beta3 subunit polymorphism in human hypertension PMID- 10817861 TI - Microbial ecology of the arbuscular mycorrhiza. AB - Arbuscular mycorrhizal (AM) fungi interact with a wide variety of organisms during all stages of their life. Some of these interactions such as grazing of the external mycelium are detrimental, while others including interactions with plant growth promoting rhizobacteria (PG PR) promote mycorrhizal functioning. Following mycorrhizal colonisation the functions of the root become modified, with consequences for the rhizosphere community which is extended into the mycorrhizosphere due to the presence of the AM external mycelium. However, we still know relatively little of the ecology of AM fungi and, in particular, the mycelium network under natural conditions. This area merits attention in the future with emphasis on the fungal partner in the association rather than the plant which has been the focus in the past. PMID- 10817862 TI - Determining replication for discrimination among microbial communities in environmental samples using community-level physiological profiles. AB - A statistical approach was employed to assess microbial community variability within a single 4-l water sample and among 10 independent 4-l water samples from a single location using community-level physiological profiles. Power calculations demonstrated that duplicate analyses could distinguish between two different locations at two times during the year. Variability associated with replicates from a single container or from different containers was nearly the same for the two sites examined. Duplicate assays (one from each of two independent samples) were sufficient to resolve the between-site differences at alpha=0.05 with power exceeding 0.95 at both times of the year. Similar methods are recommended to determine the appropriate number of replicates for environments of interest. PMID- 10817863 TI - Interactions between proteolytic and non-proteolytic Pseudomonas fluorescens affect protein degradation in a model community. AB - The metabolic interactions between proteinase-producing bacteria and other members of bacterial communities are poorly investigated, although they are important for the understanding of structure-function relationships in complex ecosystems. We constructed simple model communities consisting of proteolytic and non-proteolytic Pseudomonas fluorescens strains to identify relevant interactions and to assess their specific significance during the mobilization of protein for growth. The proteolytic or non-proteolytic model communities were established by co-inoculating proteolytic or proteinase-deficient Tn5-mutants of P. fluorescens strain ON2 with the non-proteolytic reporter strain DF57-N3 that expresses bioluminescence in response to nitrogen limitation. The growth medium was composed such that growth would be nitrogen limited in the absence of proteolytic activity. In the proteolytic communities data on growth and nitrogen availability showed that the protein hydrolysates were available to both the proteolytic and the non-proteolytic strain. Competition between these strains profoundly affected both growth and proteinase production. Hence, the mobilization of protein was closely coupled to the competitive success of the proteolytic strain. These findings provide new insight into the metabolic interactions that occur when protein is degraded in mixed bacterial communities. PMID- 10817864 TI - Physiological adaptation to low temperatures of strains of Rhizobium leguminosarum bv. viciae associated with Lathyrus spp.(1). AB - Strains of Rhizobium leguminosarum bv. viciae, isolated from the legume species Lathyrus japonicus and Lathyrus pratensis in northern Quebec (Canada), showed different capacities for growing at low temperature. In the present study, we investigated some mechanisms related to cold adaptation. Two cold-adapted strains (psychrotrophs) were compared to a poorly adapted strain and to a cold-sensitive strain (reference strain) for freezing survival, protein induction and fatty acid composition under low temperature. Following cold shocks (25 degrees C to 10, 5 and 0 degrees C), a common 6.1-kDa CSP (cold shock protein) was induced in all strains, but the total number of CSPs synthesized at 0 degrees C was higher in cold-adapted strains than in the cold-sensitive strain. The synthesis of CAPs (cold acclimation proteins) was observed under continuous growth at 5 degrees C in all three strains capable of growth at this temperature. Levels of survival after 24 h at -80 degrees C where higher in cold- (79%) and poorly adapted (64%) strains than in the cold-sensitive strain (33%), but a 2-h acclimation period at 5 degrees C before freezing doubled the survival of the cold-sensitive strain. Low temperature conditions affected similarly the fatty acid composition of all strains, regardless of their cold adaptation level. The proportion of unsaturated fatty acids increased significantly with the lowering of growth temperature from 25 to 5 degrees C, but showed a tendency to decrease after a cold shock from 25 to 5 degrees C. A specific unsaturated fatty acid, cis-12 octadecanoic acid, was produced during growth at 5 degrees C. The unsaturated cis-vaccenic acid was the principal component under all conditions. The cold adaptation trait was weakly reflected in symbiosis with the agronomic legume, Lathyrus sativus, with which one cold-adapted strain showed a slightly higher nitrogenase activity and shoot dry matter yield than a commercial strain under a sub-optimal temperature regime. PMID- 10817865 TI - Mathematical estimations of hyper-ammonia producing ruminal bacteria and evidence for bacterial antagonism that decreases ruminal ammonia production(1). AB - Mixed ruminal bacteria (MRB) from cattle fed hay produced ammonia from protein hydrolysate twice as fast as MRB from cattle fed mostly grain, and a mathematical model indicated that cattle fed hay had approximately four-fold more hyper ammonia-producing ruminal bacteria (HAB). HAB had a high maximum velocity of ammonia production (V(max)) and low substrate affinity (high K(m)), but simulations indicated that only large changes in V(max) or K(m) would cause a large deviation in HAB numbers. Some carbohydrate-fermenting ruminal bacteria produced ammonia at a slow rate (CB-LA), but many of the isolates had almost no activity (CB-NA). The model indicated that the ratio of CB-LA to CB-NA had little impact on HAB numbers. Validations based on predicted ratios of HAB, CB-LA and CB NA over-predicted the specific activity of ammonia production by MRB, but co culture incubations indicated that washed MRB from cattle fed grain could inhibit HAB. Because autoclaved MRB had virtually no effect on HAB and the incubations were always carried out at pH 7.0, the inhibition was not simply a chemical effect (e.g. low pH). PMID- 10817866 TI - Monitoring impact of in situ biostimulation treatment on groundwater bacterial community by DGGE. AB - Changes in bacterial diversity during the field experiment on biostimulation were monitored by denaturing gradient gel electrophoresis (DGGE) analysis of PCR amplified 16S rDNA fragments. The results revealed that the bacterial community was disturbed after the start of treatment, continued to change for 45 days or 60 days and then formed a relatively stable community different from the original community structure. DGGE analysis of soluble methane monooxygenase (sMMO) hydroxylase gene fragments, mmoX, was performed to monitor the shifts in the numerically dominant sMMO-containing methanotrophs during the field experiment. Sequence analysis on the mmoX gene fragments from the DGGE bands implied that the biostimulation treatment caused a shift of potential dominant sMMO-containing methanotrophs from type I methanotrophs to type II methanotrophs. PMID- 10817867 TI - The microbial food web along salinity gradients. AB - The microbial food web was studied along a gradient of salinity in two solar salterns used for the commercial production of salt. The different ponds in the salterns provide a wide range of ecosystems with food webs of different complexities. Abundance of prokaryotes, cell volume, prokaryotic heterotrophic production, chlorophyll a, abundance of heterotrophic flagellates, ciliates and phytoplankton were determined in several ponds in each saltern. Increases in salinity resulted in a progressive reduction in the abundance and number of different groups of eukaryotic microorganisms present, but an increase in biomass of prokaryotes. Maximal activity of both phyto- and bacterioplankton was found at a salinity of around 100 per thousand, where there was also a maximum in chlorophyll a concentration. Growth rates of heterotrophic prokaryotes decreased with increasing salinity. Bacterivory disappeared above 250 per thousand salinity, whereas other loss factors such as viral lysis appeared to be of minor importance throughout the gradient [Guixa-Boixereu et al. (1996) Aquat. Microb. Ecol. 11, 215-227]. PMID- 10817868 TI - Comparative analysis shows that bacterivory, not viral lysis, controls the abundance of heterotrophic prokaryotic plankton. AB - Empirical models derived from literature data were used to compare the factors controlling prokaryotic abundance (PN) and prokaryotic heterotrophic production (PHP) in solar salterns. These empirical relationships were generated as multiple linear regressions with PN or PHP as dependent variables, while the independent variables were chosen to reflect the likely sources of organic matter, inorganic nutrients and temperature. These variables were then measured in solar salterns and the predictions made by the general relationships were compared to actual saltern values of PN and PHP. Saltern ponds of salinity higher than 100 per thousand departed significantly from the general relationships, while the ponds of salinity lower than 100 per thousand fitted well within the range of values predicted by the general models. The most likely explanation for the discrepancy of the former was the absence of bacterivory. This hypothesis was tested with data from other very different aquatic systems: karstic lakes with anaerobic hypolimnia and two marine areas in the Mediterranean and the Southern Ocean. The anoxic regions of karstic lakes departed significantly from the predictions of the general model, while the oxic layers conformed to the predictions. As in the case of salterns, this difference could be explained by the presence of significant predation in the oxic, but not in the anoxic, layers of these lakes. Finally, two marine areas with similar predation pressure on prokaryotes but very different impacts of viral lysis were tested. In all cases, PN values conformed to the predictions, suggesting that lysis due to viruses is not the main factor controlling PN in aquatic systems, which is more likely to be determined by the balance between bacterivory and resource supply. The present work also demonstrates the usefulness of empirical comparative analyses to generate predictions and to draw inferences on the functioning of microbial communities. PMID- 10817869 TI - Quantitative analysis of ammonia oxidising bacteria using competitive PCR. AB - Culture-based methods for enumeration, such as most probable number (MPN) methodologies, have proved inefficient due to difficulties in the isolation and cultivation of ammonia oxidising bacteria in the laboratory. Biases are associated with the isolation of bacteria in selective media and organisms cultivated in the laboratory may not be truly representative of those in the environment. In this study, we developed a competitive PCR (cPCR)-based method based on the amplification of 16S rRNA genes specific for the beta-subgroup proteobacterial ammonia oxidising bacteria for enumeration of these organisms. Populations in both agricultural soils and estuarine sediments were quantified by traditional MPN and by cPCR. The numbers of ammonia oxidisers for both sample types were significantly underestimated by conventional MPN and were 1-3 orders of magnitude lower than those obtained by cPCR. Higher numbers of ammonia oxidisers found in fertilised plots in agricultural soils by the cPCR technique were not observed in MPN estimates. It was necessary to construct a separate standard curve for each sample type as differences in DNA extraction, quantity and purity had a significant bearing on the ease of PCR of both competitor and target DNA. PMID- 10817870 TI - Cancer and laparoscopy, experimental studies: a review. AB - OBJECTIVE: To review the experimental studies on laparoscopy and cancer and to propose guidelines for the clinical management of gynecologic cancer. METHODS: The literature in MEDLINE was searched from January 1992 to December 1998 using the terms 'cancer', 'laparoscopy' and 'experimental or animal study'. Cross referencing identified additional publications. Abstracts and letters to the editor were excluded. All the relevant papers were reviewed. RESULTS: Depending on the model used, controversial results have been reported on the incidence of trocar site metastasis when comparing CO(2) laparoscopy and laparotomy. In contrast, the following conclusions can be proposed: (i) tumour growth after laparotomy is greater than after endoscopy; (ii) tumour dissemination is worse after CO(2) laparoscopy than after laparotomy; (iii) some of the disadvantages of CO(2) laparoscopy may be treated using local or intravenous treatments or avoided using other endoscopic exposure methods, such as gasless laparoscopy. CONCLUSIONS: The laparoscopic treatment of gynecologic cancer has potential advantages and disadvantages, and may only be performed in prospective clinical trials. The risk of dissemination appears high when a large number of malignant cells are present. Adnexal tumours with external vegetations, and bulky lymph nodes should be considered as contra-indications to CO(2) laparoscopy. PMID- 10817871 TI - Diagnosis of urinary leak following abdominal total hysterectomy using renal scintigraphy. AB - Surgical trauma to the urinary system is a relatively rare complication following gynecological surgery. A case of urinary leak from rupture of the bladder following abdominal hysterectomy was diagnosed by Tc-99m-DTPA renal scintigraphy and confirmed by direct radio-isotopic cystography. Renal scintigraphic techniques should be very helpful in early diagnosis of surgical damage to the urinary tract. PMID- 10817872 TI - A retrospective comparison of water births and conventional vaginal deliveries. AB - The aim of this study was to document the practice of water births and compare their outcome and safety with normal vaginal deliveries. A retrospective case control study was conducted over a five year period from 1989 to 1994 at the Maternity Unit, Rochford Hospital, Southend, UK. Three hundred and one women electing for water births were compared with the same number of age and parity matched low risk women having conventional vaginal deliveries. Length of labour; analgesia requirements; apgar scores; maternal complications including perineal trauma, postpartum haemorrhages, infections; fetal and neonatal complications including shoulder dystocias; admissions to the Special Care Baby Unit, and infections were noted. Primigravidae having water births had shorter first and second stages of labour compared with controls (P<0.05 and P<0.005 respectively), reducing the total time spent in labour by 90 min (95% confidence interval 31 to 148). All women having water births had reduced analgesia requirements. No analgesia was required by 38% (95% confidence interval 23.5 to 36.3, P<0.0001) and 1.3% requested opiates compared to 56% of the controls (95% confidence interval 46. 3 to 58.1, P<0.0001). Primigravidae having water births had less perineal trauma (P<0.05). Overall the episiotomy rate was 5 times greater in the control group (95% confidence interval 15 to 26.2, P<0.0001), but more women having water births had perineal tears (95% confidence interval 6.6 to 22.6, P<0.001). There were twice as many third degree tears, post partum haemorrhages and admissions to the Special Care Baby Unit in the controls, although these differences were not significant. Apgar scores were comparable in both groups. There were no neonatal infections or neonatal deaths in the study. This study suffers from many of the methodological problems inherent in investigation of uncommon modes of delivery. However, we conclude that water births in low risk women delivered by experienced professionals are as safe as normal vaginal deliveries. Labouring and delivering in water is associated with a reduction in length of labour and perineal trauma for primigravidae, and a reduction in analgesia requirements for all women. PMID- 10817873 TI - The evaluation of determinants of early postpartum low mood: the importance of parity and inter-pregnancy interval. AB - OBJECTIVES: The aim of this study is to determine the extent of early postpartum low mood and possible relevant variables in our population. STUDY DESIGN: Depressive symptoms were examined in a sample of 85 women in the first week of postpartum period using Beck Depression Inventory (BDI). The relevant sociodemographic variables were tested by multiple logistic regression analysis. RESULTS: The rate of postpartum low mood (BDI score >10) was found 35.2%. High parity, long marriage period and low education level were all significantly associated with low mood in the postpartum first week. When logistic regression analysis was used to eliminate confounding effect of the parameters; grandmultiparity, short inter-pregnancy interval and low educational level were found to have important effect on postpartum low mood. CONCLUSION: Pregnant women with risk factors for postpartum low mood such as grandmultiparity, short inter pregnancy interval and low educational level should be identified early during antenatal period. Educational and social support should be given to those in such a risk. We believe that effective family planning programs and improvement of women's educational level are important preventive factors for postpartum low mood and subsequently developing depression. PMID- 10817874 TI - Postpartum curettage in patients with HELLP-syndrome does not result in accelerated recovery. AB - OBJECTIVE: Our purpose was to examine the impact of immediate postpartum curettage on the recovery of patients with HELLP-syndrome (hemolysis, elevated liver enzymes, and low platelets). SUBJECTS: Between January 1994 and July 1997 all patients who presented with HELLP-syndrome in our institution underwent immediate postpartum curettage (n=24). Their outcome was compared with the recovery of women with HELLP-syndrome who were delivered without postpartum curettage between 1987 and 1993 (n=20). Clinical and laboratory data were analyzed. RESULTS: No significant difference could be found between both groups in terms of normalization of serum glutamic-oxaloacetic transaminase (GOT), serum glutamic-pyruvic transaminase (GPT), lactic dehydrogenase (LDH), and quantitative platelet count. Postpartum hospitalization time was identical in both groups. CONCLUSION: In our retrospective study no benefit is achieved by immediate postpartum curettage in patients with HELLP-syndrome. In order to eliminate the possible bias of retrospective analysis, we now plan a randomized study to further investigate the impact of immediate postpartum curettage. PMID- 10817875 TI - Screening for gestational diabetes: variation in guidelines. AB - OBJECTIVE: To compare published guidelines concerning screening for gestational diabetes. STUDY DESIGN: Systematic search and comparative analysis of published guidelines. Appraisal of guidelines quality. Simulation analysis. RESULTS: Ten published guidelines proposed either universal screening (5), selective screening (3) or screening when clinically indicated (2). Variations of testing schedules and blood glucose thresholds were observed. The quality of the published guidelines was low, on average 22 (8-51) percentage points on the assessment scale. These differences would have led to large variations in the number of patients to be screened. CONCLUSIONS: Large variations between guidelines have been observed which would translate in large practice variations, if the guidelines were systematically applied. These variations are partially explained by the absence of definite evidence that universal or selective screening for gestational diabetes do more good than harm on infant and maternal health. The methodology of developing guidelines should be more evidence based, systematic and explicit. PMID- 10817876 TI - Overestimation and underestimation of labor pain. AB - OBJECTIVE: To define factors affecting the ability of the medical staff to estimate the level of pain during labor. METHODS: The study population consisted of 255 consecutive women. All parturients were asked to rate their pain level, using a visual analog scale. At the same time, the caregivers estimated the degree of pain as was exhibited by the parturients, using the same scale. Patients whose pain level was either overestimated or underestimated were compared to patients whose pain was correctly estimated. RESULTS: In about half of the women (50.6%) enrolled in our study the level of pain was estimated correctly by the caregivers, while similar proportions of women had their pain level overestimated (24.3%) and underestimated (25.1%) by the caregivers. While the majority (54.5%) of patients who were in their second or third deliveries were equally estimated by the personnel with regard to their pain intensity levels, most of the grandmultiparous women had their labor pain underestimated by the medical staff. (57.1%, P<0.001). None of the caregivers had more than five children. Although the mostly secular medical staff could properly estimate the pain levels of most secular patients (52.3%), the labor pain of 44% of the religious parturients was underestimated (P=0.003). The ability to estimate properly the patients' suffering was found to be unaffected by age, family status, educational level, presence of companion during labor and participation in childbirth preparation course. CONCLUSION: The wider the cultural gap between the caregiver and the patient, the less accurate was the interpretation of patient's pain. PMID- 10817877 TI - Caesarean section in previously untreated acute promyelocytic leukaemia. Report of two patients. AB - Acute promyelocytic leukaemia (APL) is characterised by a life-threatening hemorrhagic diathesis which is attributed to a DIC-like coagulopathy. This report describes the problems of childbirth in two patients with untreated APL. It is concluded that caesarean section can be performed without major complications. A prerequisite is an active treatment of the coagulopathy and a close collaboration between the obstetrician and the haematologist. PMID- 10817878 TI - Regional trend variations in infant mortality due to perinatal conditions in the Netherlands. AB - CONDENSATION: In the Netherlands, regional variations in trends in infant mortality due to perinatal conditions (1984-1994) exist, which could not be explained by health care characteristics (i.e., place or supervision of delivery and the presence of specialised neonatal care). The only sociodemographic factor that showed a consistent correlation with mortality was the percentage of Roman Catholic inhabitants of a region. OBJECTIVE: To describe and explain regional variations in trends in infant mortality due to perinatal conditions. STUDY DESIGN: A mixed (geographical and temporal) ecological design has been used. Infant mortality due to perinatal conditions was defined as mortality in the first year of life caused by diseases of the newborn period (chapter XV of the ICD-9). Trends in sex-adjusted mortality for the period 1984-1994 as well as mortality levels at the start of this period were calculated using log linear regression. Linear regression was used to examine the association between mortality trends and starting levels on the one hand and both health care and sociodemographic factors on the other. RESULTS: Statistically significant variations in mortality trends were found between regions. The trends in the two Southern regions were found to deviate significantly from the national trend. No strong association was found between mortality and each of the health care factors (i.e. place and/or supervision of delivery and the presence of specialised neonatal care). The only sociodemographic factor that showed consistent results was the percentage of Roman Catholic inhabitants of a region: A higher percentage in 1985 was associated with a higher mortality in 1985 and a stronger mortality decline during the period 1984-1994. This association could not be explained by parity or the age of the mother. CONCLUSIONS: Regional differences in trends in infant mortality due to perinatal conditions in the Netherlands could not be explained by variations in health care factors. This is an important finding as the Dutch system of obstetric care, that includes a considerable number of home deliveries, has been subject to much debate. Further research that includes other causes of death and determinants is needed to unravel the causes of the trend variations. PMID- 10817879 TI - Neuroendocrine tumors of the uterine cervix. Clinicopathologic study of five patients. AB - Four main clinicopathologic features of neuroendocrine tumors (NETs) of the cervix may be stressed: primary diagnosis at an advanced stage, early nodal metastasis even for low disease, early failure of appropriate local treatment (surgery and/or radiation therapy) and aggressive clinical treatment. Five patients with NET of the uterine cervix (small cell carcinoma type) are reported (one stage I, two stages II, one stage III and one stage IV). One patient was treated by surgery combined with radiation therapy, one by surgery combined with chemotherapy and one by surgery with radiation therapy and chemotherapy. Two patients received radiation therapy alone. Three early stage patients are alive with no evidence of disease 8, 26 and 41 months after diagnosis. The two patients with advanced stage died of disease, 3 and 12 months respectively, after diagnosis. Combination chemotherapy (cisplatin and etoposide) is warranted in disseminated NETs. Neoadjuvant or adjuvant chemotherapy should be combined with radiation therapy and surgery even in early stages. PMID- 10817880 TI - Sensitivity and specificity of ultrasound indices of ovulation in spontaneous cycles. AB - OBJECTIVE: Evaluation of sensitivity and specificity of 4 ultrasound indices of ovulation. STUDY DESIGN: Multicenter collaborative study of 794 abdominal and transvaginal ultrasound scanning of ovaries performed during 271 cycles in 107 normally fertile women. Comparison of sensitivities and specificities of indices using McNemar test. RESULTS: The sensitivity and specificity of the indices were 84 and 89.2, respectively, for disappearance or sudden decrease in follicle size; 38.4 and 79.7 for appearance of ultrasonic echoes in the follicle; 61.6 and 87.1 for irregularity of follicular walls; 71.0 and 88.2 for appearance of free fluid in the cul-de-sac of Douglas. CONCLUSION: Ultrasonic echoes had a significantly lower sensitivity (P<0.001) and specificity (P<0.01) than other indices. PMID- 10817881 TI - Comparison of protective media and freezing techniques for cryopreservation of human semen. AB - OBJECTIVE: To evaluate the influence of cryopreservation medium and freezing thawing techniques on human sperm motility and morphology. STUDY DESIGN: 63 semen samples were obtained from 39 donors to the artificial insemination programme. Possible effects of the sperm dilution with cryomedium on the motility were examined 10 min after exposure of 24 high initial quality semen samples to TEST yolk ?zwitterion-citrate-egg yolk extender containing TES [N-Tris (hydroxymethyl) methylaminoethane sulfonic acid] and Tris [(hydroxymethyl) aminomethane]? and human sperm preservation medium (HSPM). Post-thaw sperm motility from 24 frozen semen samples was examined comparing the cryoprotective efficacy of TEST-yolk and HSPM following different freezing techniques (vapour freezing, fast programmable freezing and slow programmable freezing). The relationship of sperm morphology to the effects of freezing was investigated on 39 semen samples following different freezing techniques. Post-thaw sperm motility from 39 frozen semen samples was compared among three groups divided according to the percentage of morphologically normal cells (<40, 40-50 and >50%) in fresh semen. RESULTS: Exposure of spermatozoa to cryomedia for 10 min at room temperature significantly reduced motility in TEST-yolk treatment group for 9% and in HSPM treatment group for 18% (P<0.01). The recovery of motile sperms (mean+/-standard deviation) was 49+/-15.7, 43+/-15.2 and 52+/-16.8% when TEST-yolk was used and 34+/-17.8, 32+/ 18.2 and 50+/-13.6% when HSPM was used as a cryopreservative following vapour freezing, and fast and slow programmable freezing, respectively. Following vapour freezing and also following fast programmable freezing, the recovery of motile sperm was significantly higher (P<0.05) after addition of TEST-yolk medium than after addition of HSPM. Post-thaw motility of the sperm cryopreserved in HSPM showed significant differences (P<0.05) after three different freezing techniques. The recovery of motile sperms was 57+/-26.4, 38+/-8.6 and 38+/-17.3% in groups with >50, 40-50 and <40% morphologically normal cells, respectively. The percentage of morphologically normal spermatozoa was reduced 8% after vapour freezing and 6 and 3% after fast and slow programmable freezing, respectively. The results were statistically analysed using SAS/STAT software. CONCLUSIONS: Slow programmable freezing was superior to vapour freezing and fast programmable freezing as a method for sperm cryopreservation. However, none of these methods of freezing had discernible effects on sperm morphology. Motility of spermatozoa decreased due to the exposure of semen to cryomedium. TEST-yolk was a superior cryomedium to HSPM. Fresh semen with more than 50% of morphologically normal cells showed the best recovery of motile cells after freezing and thawing. PMID- 10817882 TI - Severe bradycardia and bradypnea following vaginal oocyte retrieval: a possible toxic effect of paracervical mepivacaine. AB - We report a case of a patient with a history of heart conduction disease, symptom free and without treatment in the last years, who experienced a severe cardiac complication associated with in vitro fertilization (IVF) with vaginal oocyte retrieval (VOR). Eighty-five minutes after the VOR a severe bradycardia and bradypnea occurred, requiring an emergency application of a pacemaker. Presumably the condition occurred because of a toxic effect of the 400 mg of mepivacaine administered paracervically. It is concluded that in the paracervical anesthesia in the IVF cycles the therapeutic range should be scrupulously followed in patients with heart condition. PMID- 10817883 TI - Inversion of uterus during cesarean section. AB - Acute inversion of the uterus is a rare complication during cesarean section. We describe one such case in which diagnosis was made immediately and reversion was performed within few minutes. A high index of suspicion and prompt management can prevent further complications. PMID- 10817884 TI - Prenatal diagnosis of a fetal abdominal cyst. AB - Hepatic liver cysts are rare and few cases have been described in the literature. Initially, these cysts are detected by a prenatal ultrasound as abdominal, and consequently, the final diagnosis of hepatic cysts is done during the postnatal period. We present a case of a giant hepatic cyst found incidently during a routine ultrasound examination. The diagnosis and management of a fetal intraabdominal cyst is discussed. PMID- 10817885 TI - Urinomas as a complication of iatrogenic ureteric injuries in gynecological surgery. AB - We examined the incidence of ureteric injuries in relation to gynecologic operations. In 5240 gynecological operations, eighteen (0.34%) cases of ureteric injuries and four cases of urinomas were found. We present the four cases of ureteric injuries that were found with postoperative development of urinomas. PMID- 10817886 TI - Transient myocardial ischemia may occur following subendometrial vasopressin infiltration. AB - A case of transient myocardial ischemia following subendometrial vasopressin infiltration in intractable intra-operative postpartum bleeding due to placenta accreta is described. In our experience, the rate of this side effect is one in 14 patients (rate of 7.1%). We believe that the benefits of the treatment outweigh the risks, since the uterus was saved in all 14 patients. Nevertheless, this case emphasises that extreme precaution is needed with subendometrial vasopressin infiltration. It should be emphasised that the needle must not be within a blood vessel because intravascular injection of vasopressin solution can precipitate acute arterial hypertension, bradycardia and even death. We suggest that local vasopressin infiltration into the placental site is indicated in cases of intractable bleeding at cesarean section after other conventional obstetric and pharmacological maneuvers have failed. PMID- 10817887 TI - A large yolk sac may be important in the early diagnosis of gestational trophoblastic disease: a case report. AB - A large yolk sac in a deformed shape gestational sac was detected by ultrasonography in an 8 week pregnant woman. The disappearance of the yolk sac and the gestational sac, enlargement of the uterine cavity and a heterogeneous appearance similar to early gestational trophoblastic disease were recognized when ultrasonographic examination was performed 2 weeks later. After uterine evacuation, partial hydatidiform mole was diagnosed by histopathological evaluation of the curettage material. In our study, the role of the large yolk sac in predicting trophoblastic disease is discussed. PMID- 10817888 TI - Multimodal cancer chemotherapy during the first and second trimester of pregnancy: a case report. AB - This paper reports treatment with combined chemotherapy during pregnancy. A 39 year-old woman with breast cancer was given adjuvant chemotherapy including cyclophosphamide, methotrexate and 6-fluorouracil from the 6th to the 24th week of gestation. The possibility of teratogenic effects on the fetus was explained to the patient however she refused to terminate the pregnancy. A 30-week male infant with only a minor malformation was delivered. The authors reviewed the literature regarding chemotherapeutic agents given during the first trimester of pregnancy. Most cytotoxic drugs have teratogenic effects on experimental animal subjects. However, actual data on human fetuses are sparse because of the variety of therapeutic regimens and the rarity of administering chemotherapy during pregnancy. The long-term effects of exposure to cytotoxic drugs in utero, needs further research. PMID- 10817889 TI - Lyme borreliosis as a cause of facial palsy during pregnancy. AB - The medical history of a pregnant woman in whom the initial pattern of complaints suggested hyperemesis gravidarum is described. After about 18 days the patient developed left facial palsy. Repeated tests eventually confirmed the diagnosis of neuroborreliosis. The problems concerning diagnostics, therapy and the possible complications of Lyme borreliosis during gestation are described. PMID- 10817891 TI - Long-term outcome of 17 cases of large-giant posterior fossa aneurysm. AB - Long-term outcome of 17 patients who harbored a large or giant aneurysm of posterior fossa was summarized. The anatomical distribution of aneurysms included eight cases of basilar artery (BA) bifurcation aneurysms, three cases of BA trunk aneurysms, and six cases of vertebral artery (VA) aneurysms. Eight patients received surgical or endovascular treatment for their lesion. The clinical outcome was good recovery in six, moderate disability in one, and vegetative state in one case, respectively. The other nine patients were followed conservatively. Four of them had fatal aneurysmal rupture, and another two patients suffered from aggravation of pre-existing symptoms related to their aneurysm. Only three patients remain intact. Comparison of the radiographic parameters between those who bled and those who did not bleed revealed that those with subsequent rupture had significantly higher rate of aneurysmal thrombus and had a trend for larger diameter of the aneurysm. Although more aggressive and multidisciplinary measure should be taken to these patients to improve their long term outcome, our results showed the limitation of treatment for these patients in the present era at the same time. The patients with broad neck BA bifurcation aneurysm in which efferent vessels were incorporated into aneurysmal dome, and those with fusiform, giant BA trunk aneurysm with thrombus were the least amenable to treatment in our series. PMID- 10817892 TI - Intrasellar tuberculoma--an enigmatic pituitary infection: a series of 18 cases. AB - OBJECTIVE: Intrasellar tuberculomas are rare and only few case reports have been described in the literature. We report a series of 18 cases of histologically proven intrasellar tuberculomas, which, to the best of our knowledge, is the largest series in the English literature. METHODS: A total of 1143 pituitary lesions, between 1984 and June 1999, were operated for various reasons in our institute. Of these, 18 cases were histopathologically proven intrasellar tuberculomas. The clinical profile was reviewed in detail. Radiological data and histopathological slides were also reviewed. RESULTS: The age ranged from 8 to 43 years (average 23.6 years) with a female preponderance. The duration of symptoms varied from 15 days to 2 years (average 4 months), the most common symptoms being headache followed by decrease or loss of vision. Five patients had features of pan-hypopituitarism whereas three had raised prolactin (PRL) levels. In six patients, both sella as well as sphenoid sinus were involved. In one patient the lesion was extending from the sella over the clivus. Clinically as well as radiologically, these lesions were mistaken for pituitary adenomas except for one case where tuberculoma was suspected on imaging. In three patients, there was past history of pulmonary tuberculosis, in one patient of tuberculous meningitis, and in one patient, of spondilytis of the spine. In one patient there was cervical lymphadenopathy along with features of acromegaly (also proved by high levels of serum growth hormone) and radiology revealed a pituitary pathology. Microscopic examination of the excised lesion revealed a composite lesion consisting of a pituitary adenoma and tuberculoma, which has not been documented in literature to date. One patient died during the hospital stay. All the other patients were put on antitubercular chemotherapy following surgery and had good outcomes. CONCLUSION: Intrasellar tuberculomas are rare. These may be suspected in female patients especially if radiological imaging shows involvement of paranasal sinuses and pituitary fossa along with thickening of pituitary stalk. Simultaneous involvement of clivus may also be an additional feature. The incidence of pituitary tuberculosis is likely to increase with a rise in the incidence of AIDS. PMID- 10817893 TI - Initial hyperglycemia as an indicator of severity of the ictus in poor-grade patients with spontaneous subarachnoid hemorrhage. AB - An association between hyperglycemia and outcome in spontaneous subarachnoid hemorrhage (SAH) has been sporadically reported. Our hypothesis was that hyperglycemia is a sign of central metabolic disturbance linked with specific appearances on computerized tomography (CT) scans reflecting different degrees of corresponding brain injury. The admission plasma glucose level, initial CT findings, and outcome after 6 months were analysed in a cohort of 99 patients with SAH in Hunt & Hess Grade IV or V. The CT scans were quantitatively assessed for subarachnoid blood, intracerebral hematoma, intraventricular hemorrhage, hydrocephalus, midline shift and compression of the perimesencephalic cisterns. These findings were combined to determine a three-point CT severity score. All patients showed elevated (>5.8 mmol/l) plasma glucose levels on admission. Mortality among 33 patients with glucose concentration below 9.0 mmol/l was 33.3%, 71.1% for the 45 patients with glucose level between 9.0 and 13.0 mmol/l, and 95.2% for the 21 patients with concentration above 13.0 mmol/l (P<0.0001). Glucose level was higher in Grade V than in Grade IV patients (mean+/-SD) (11.8+/ 3.2 vs 9.8+/-2.9 mmol/l; P=0.0012). Patients with mild CT findings (n=10) had the lowest glucose level (8.9+/-1.8 mmol/l; P=0.0082), whereas patients with severe findings (n=56) had the highest glucose (11.4+/-3.5 mmol/l; P=0.011). Despite association with clinical grade and extent of CT findings, logistic multiple regression revealed the admission plasma glucose level to be an independent prognosticator of outcome. The prognostic potential of the initial plasma glucose level may be beneficial in management protocols of poor-grade SAH patients. PMID- 10817894 TI - Acute effects of thalamotomy and pallidotomy on regional cerebral metabolism, evaluated by PET. AB - The subacute effect of thalamotomy and pallidotomy on regional cerebral metabolism was studied by means of Positron Emission Tomography (PET). In this way we aimed to identify the pattern of functional deafferentiation following a specific lesion in the basal ganglia. The cerebral distribution of 2-[18F]fluoro 2-deoxy-D-glucose (FDG) uptake at 1-2 weeks after operation was compared with the uptake before operation. Analysis of the changes was done by statistical parametric mapping (SPM). Thalamotomy resulted in a reduction of FDG uptake in predominantly the lateral prefrontal- and the parietal cortex, whereas pallidotomy affected only uptake in the (pre)frontal cortex. The absence of change in the primary sensory-motor cortex after either surgical procedure may suggest that, in man, the motor portions of the thalamus exert a predominantly indirect influence on the human motor cortex. PMID- 10817895 TI - Posterior fossa decompression in syringomyelia associated with a Chiari malformation: a retrospective analysis of 22 patients. AB - BACKGROUND: The technique of posterior fossa decompression, nowadays preferred by most neurosurgeons as the first surgical option in the treatment of hindbrain associated syringomyelia, was reintroduced in our center in 1989. We present a retrospective analysis of 22 patients with this pathology who underwent the procedure since then. METHODS: In all patients diagnosis was made by MRI of the craniospinal junction. The operation consisted of a suboccipital craniectomy, removal of the posterior arc of C1, laminectomy of C2 or C3 when necessary and a wide duraplasty at the end. The intradural manipulations (opening of the arachnoid membrane, coagulation or resection of the tonsils) were not uniform. Postoperatively short- and long-term clinical outcome and MRI findings were assessed. RESULTS: Sixteen out of 21 patients (76%) experienced an improvement in the early follow-up period. In the late follow-up period 13 out of 19 patients (68%) were improved, whereas five patients (26%) experienced a marked deterioration. There was no unequivocal effect on all symptoms and signs. Postoperative MR images showed a favorable result in 16 out of 20 patients (80%), consisting of syrinx collapse or reduction of the syrinx diameter. CONCLUSIONS: We conclude that decompression of the posterior fossa is a safe procedure with a considerable chance of clinical improvement. Although total syrinx collapse is not as frequently seen as in syrinx shunting procedures, the clinical outcome may be better. Moreover, there seemed to be no unequivocal correlation between clinical outcome and postoperative syrinx size in the present study. PMID- 10817896 TI - Congenital fibre type disproportion a time-locked diagnosis: a clinical and morphological follow-up study. AB - This is the first report that describes the clinical and histological findings in a patient suffering from pure congenital fibre type disproportion (CFTD), who had two biopsies with an interval of 16 years. Additionally, we compared the clinical signs and symptoms of the present case to those of 35 CFTD cases reviewed from the literature. From this we conclude that smallness of type I fibre diameter is not a characteristic feature during the course of pure CFTD. This suggests that CFTD is a time-locked diagnosis. PMID- 10817897 TI - Frontal-executive dysfunction in early onset cerebellar ataxia of Holmes' type. AB - We report the case of a 29-year-old male patient with cerebellar ataxia of Holmes' type. The combination of progressive cerebellar ataxia and hypogonadotrophic hypogonadism is a rare distinctive syndrome which was first described by Holmes in 1907. Early diagnosis is desirable because replacement of testosterone may allow normal sexual development. MRI showed severe combined superior vermian and cerebellar hemisphere atrophy. Comprehensive neuropsychological testing pointed to a more widespread cerebellar mediated functional CNS involvement in the earlier stages of this ataxic syndrome than previously described in mentally not retarded subjects. PMID- 10817898 TI - Familial brainstem glioma. AB - Two young males with a high degree of consanguinity presented with similar brainstem gliomas. As the patients were raised separately, their presentation with similar tumors suggests a genetic basis for the occurrence of these gliomas. PMID- 10817899 TI - Lower motor neuron disease associated with myelofibrosis. AB - We present a patient who has signs pointing to the involvement of lower motor neurons and myelofibrosis. To our knowledge, unlike lymphoproliferative disorders, co-occurrence of myelofibrosis and lower motor neuron disease (MND) has not been reported so far. A 64-year-old male patient was admitted to our hospital with the complaint of painful cramps in his neck and forearms. On physical examination marked hepatomegaly and splenomegaly were found. On neurologic examination nasal quality of the voice and slight palatal weakness were detected. There were generalised slight weakness and atrophy in both proximal and distal muscle groups. Fasciculations were observed especially in forearm muscles and it was observed that he had been avoiding head movements because of painful muscle cramps. Deep tendon reflexes were hypoactive. Nerve conduction studies were normal. By needle electromyography, giant motor unit action potentials (amplitudes up to 8 mV), fibrillation potentials, positive sharp waves and fasciculation potentials were detected in all muscles which were investigated. A hypercellular bone marrow (100%) was determined by bone marrow biopsy. In addition to increased production of the myeloid and megakaryocytic lines, abnormal aggregation and grouping of megakaryocytes were seen. Reticular fibers were increased. He had some benefit of dyphenilhydantoin treatment given for the painful cramps in his neck and forearm muscles. Hydroxyurea treatment was started for myelofibrosis. Six months later, his general condition was better, and the painful cramps were completely resolved. No marked deterioration has been detected in neurologic examination and electromyography for 1 year. PMID- 10817900 TI - Recurring aseptic meningitis after travel to the tropics: a case of Mollaret's meningitis? Case report with review of the literature. AB - Recurrent aseptic meningitis in a 35-year-old caucasian woman is described. She had many attacks over a period of 9 years. The first attack occurred after travel in the tropics. In spite of extensive examinations no cause could be found for the recurrent attacks. Both the clinical presentation and characteristics of the cerebrospinal fluid are compatible with the diagnosis of Mollaret's meningitis. There is no known cure for this condition, although colchicine and indomethacin have been mentioned to relieve symptoms. In our patient, a treatment with indomethacin during the last attack resulted in a clear and rapid improvement of symptoms. Since this episode only mild relapses have occurred, all of which responded well to the same treatment. This case highlights the long time span in which attacks of Mollaret's meningitis can occur, and the spectacular benefit of indomethacin. PMID- 10817901 TI - Oxidative stress and antioxidative defense in cephalopods: a function of metabolic rate or age? AB - Activities of the antioxidative enzymes superoxide dismutase (SOD), catalase, glutathione peroxidase (GPX) and glutathione reductase (GR) were measured in the cephalopods Sepia officinalis and Lolliguncula brevis. Maximal enzyme activities were higher in gill tissue than in the mantle musculature of both species. Activities were generally lower in tissues of L. brevis than in S. officinalis. Comparison with other ectothermic animals showed both cephalopod species to have a low enzymatic antioxidative status despite their high metabolic rate. Furthermore, changes in antioxidative enzyme activities were measured in the cuttlefish S. officinalis with increasing age. The concentrations of malondialdehyde (MDA) and lipofuscin were determined as indicators of lipid peroxidation. Investigated animals were between 1.5 months and over 12 months old. Changes of antioxidative enzyme activities with age were not uniform. SOD and GPX activities increased with age, while catalase activity declined. In contrast, GR activity remained almost unchanged in all age groups. The low level of antioxidative defense might allow for the significant age-induced rise in MDA levels in gills and mantle musculature and for the increase in lipofuscin levels in mantle and brain tissue. It might thereby contribute to increased oxidative damage and a short life span in these cephalopods. PMID- 10817902 TI - 2',3'-cyclic nucleotide 2'-phosphodiesterase from Fusarium culmorum. AB - We describe the properties of a 2',3'-cyclic nucleotide 2'-phosphodiesterase (EC 3.1.4.16), found in Fusarium culmorum, which hydrolyzes nucleoside 2',3'-cyclic monophosphates to nucleoside 3'-phosphates. In contrast with a similar enzyme found in bacteria, the Fusarium enzyme does not exhibit nucleotidase activity and does not show a requirement for metal ions, but is inhibited by micromolar concentrations of Cu++ and Zn++, and is very stable to heat. This cyclic phosphodiesterase hydrolyzes the four major nucleoside 2',3'-cyclic monophosphates and has greater affinity for purine (Kms for Ado-2',3'-P = 0.3 mM and for Guo-2',3'-P = 0.1 mM) than for pyrimidine nucleotides (Kms for Cyd-2',3' P = 0.6 mM and for Urd-2',3'-P = 2 mM). The respective Vmax for Urd-2',3'-P; Cyd 2',3'-P; Ado-2',3'-P; and Guo-2',3' are 100:45:16:5. The efficacy of the phosphodiesterase to hydrolyze the four major 2',3' cyclic nucleotides (based on the relative values of Vmax/Km) is not significantly different. The Fusarium enzyme differs from a previously described 2',3' cyclic phosphodiesterase from Neurospora, in that it is inactive on 3',5'-nucleoside monophosphates and nucleoside 2' or 3' phosphates. PMID- 10817903 TI - Purification and characterization of a novel lectin from a freshwater cyanobacterium, Oscillatoria agardhii. AB - In the survey of 14 species of laboratory-cultured cyanobacteria for hemagglutinins, we newly detected the activity in two species, Oscillatoria agardhii, strain NIES-204, and Phormidium foveolarum, strain NIES-503. From the extract of O. agardhii, which showed the highest activity with trypsin-treated erythrocytes of rabbit, a lectin was purified to homogeneity by the combination of precipitation with (NH4)2SO4, gel filtration, hydrophobic chromatography and reverse phase chromatography. The purified lectin, designated OAA, was a monomeric protein with an apparent molecular weight of 13,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 16,000 on gel filtration. The amino acid composition was rich in glycine and acidic amino acids. The hemagglutination activity was inhibited by glycoproteins such as yeast mannan, but not by any of the monosaccharides tested. The activity was stable over a wide range of pH (4-11) and at a high temperature of 80 degrees C, and independent on the presence of divalent cations. The features of OAA resembled those of many of lectins from marine macroalgae. The sequence of amino-terminal residues of OAA was determined as ALYNVENQWGGSSAPWNEGG, which was highly homologous to those of lectins from macroalgae of the genus Eucheuma and that of a myxobacterium Myxococcus xanthus hemagglutinin. PMID- 10817904 TI - Purification and partial characterization of haloperoxidase from fresh water algae Cladophora glomerata. AB - Many haloperoxidases have been purified from diverse organisms, including lichen, fungi, bacteria, and marine algae. In this study a haloperoxidase was purified from the fresh water algae, Cladophora glomerata, by homogenization and centrifugation, ammonium sulfate fractionation, ion-exchange and gel filtration chromatography. Molecular weight was determined by SDS-PAGE and by size exclusion HPLC and found to be approximately 43 kDa. The isoelectric point was determined to be approximately 8.1 by isoelectric focusing. The UV spectrum of the peroxidase showed a strong absorbance in the Soret band indicating a heme protein, unlike vanadium-dependent haloperoxidases from marine algae. Fresh water algal haloperoxidase catalyzed the iodination of tyrosine at a pH of 3.1. This haloperoxidase also catalyzes the oxidation of guaiacol and oxidation of iodide as well as catalyzing a peroxide-dependent reaction in both the presence and absence of chloride and bromide ions. PMID- 10817905 TI - Localization of arginine vasotocin (AVT) mRNA in extrasomal compartments of magnocellular neurons in the chicken hypothalamo-neurohypophysial system. AB - In the chicken, arginine vasotocin (AVT) is produced in and secreted by magnocellular neurons in the supraoptic (SON) and paraventricular (PVN) nuclei. To test the hypothesis of axonally transported AVT mRNA, the localization of AVT mRNA within extrasomal, axonal/dendritic compartments in the chicken hypothalamo neurohypophysial system (HNS) were examined using AVT specific in situ hybridization histochemistry (ISHH) and RT-PCR. Many perikarya in the PVN and external--but none in the ventral subgroup of the SON show ISHH signals clearly extended into one or two processes, some with branching collaterals, traceable over a distance of more than 100 microns. Furthermore by using RT-PCR, AVT mRNA was detected in the median eminence and neurohypophysis representing the distal parts of the HNS, mainly consisting of axons and/or axon terminals. These observations of axonal mRNA offer new insights to the organization and function of the avian HNS. PMID- 10817906 TI - Characteristics of sperm acrosin-like activity of paddlefish (Polyodon spathula Walbaum). AB - Spermatozoa of paddlefish and sturgeon fishes (Acipenseriformes), unlike teleost fish, have an acrosome. The objectives of this study were to characterize acrosin like activity of cryopreserved sperm of paddlefish (Polyodon spathula) and to test and compare stability of paddlefish acrosin-like activity with that of lake sturgeon and bull spermatozoa. Mean acrosin-like activity of cryopreserved paddlefish sperm was 0.372 +/- 0.067 microU/10(6) spermatozoa. This activity was 79% higher in the whole semen than in spermatozoa. Highest activity was recorded at pH 8.0 and 8.5. Triton X-100, zinc ions and 4'-acetamidophenyl 4 guanidinobenzoate (AGB) inhibited the activity. Amidase activity was also inhibited by N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) and N-tosyl-L phenylalanine chloromethyl ketone (TPCK). TLCK at concentrations of 0.1 and 1.0 mM gave a significant decrease in activity of 19 and 61%, respectively. However, TPCK significantly inhibited amidase activity (by 19%) only at concentration 1.0 mM. After acidification and 60 min incubation at 4 degrees C of sperm suspensions only 4% of the activity was retained. A similar phenomenon was observed in the case of lake sturgeon but not bull sperm. These results suggest that trypsin-like activity of Acipenserid fish resembles rather fish trypsin that mammalian one. In frozen-thawed paddlefish sperm a minute chymotrypsin-like activity was also indicated, when GPNA was used as substrate. This activity amounted to 0.0415 +/- 0.0138 microU/10(6) spermatozoa and was 18% of total amidase activity. This suggests that chymotrypsin-like activity may also be present in paddlefish spermatozoa. PMID- 10817907 TI - Characterization of acetohydroxy acid synthase activity in the archaeon Haloferax volcanii. AB - Whereas the biochemistry of acetohydroxy acid synthase has been extensively studied in bacteria and eukaryotes, relatively little is known about the enzyme in archaea, the third kingdom of life. The present study biochemically characterizes acetohydroxy acid synthase activity in the halophilic archaea Haloferax volcanii. In addressing ion requirements, enzyme inhibition and antibody labeling, the results reveal that, except for its elevated salt requirements, the haloarchaeal enzyme is remarkably similar to its bacterial counterpart. PMID- 10817908 TI - Sulfurtransferases and the content of cysteine, glutathione and sulfane sulfur in tissues of the frog Rana temporaria. AB - L-cysteine desulfuration was examined in tissues of Rana temporaria, in October and January. The activities of 3-mercaptopyruvate sulfurtransferase (MPST), cystathionine gamma-lyase (CST) and rhodanese were primarily concentrated in frog liver and kidney. The values of CST and rhodanese activity, as well as sulfane sulfur compounds levels fell in the range characteristic of rat. For each of the investigated tissues changes noted in the enzymatic activities and in the level of glutathione (GSH), protein-bound cysteine (PbCys) and sulfane sulfur compounds were dependent on the month in which the determination was performed, and on the character of the tissue. In such tissues as the liver or gonads, high GSH levels and high activities of MPST (in the liver) or MPST and rhodanese (in the gonads) seemed to accompany protein biosynthesis during hibernation. PbCys, the level of which was consequently diminished in all tissues in January, compensated the absence of exogenous cysteine. A significantly reduced GSH level in the brain in January seemed to be correlated with decreased requirements of the tissue for this important natural antioxidant at diminished thyroid hormones levels in the serum and minimal oxygen consumption during the hibernation. In the kidney, the possible participation of sulfane sulfur compounds in detoxification processes requires elucidation, similarly as in protection against cellular oxidative stress at extremely low levels of GSH. PMID- 10817909 TI - Scaling effects on hypoxia tolerance in the Amazon fish Astronotus ocellatus (Perciformes: Cichlidae): contribution of tissue enzyme levels. AB - Astronotus ocellatus is one of the most hypoxia tolerant fish of the Amazon; adult animals can tolerate up to 6 h of anoxia at 28 degrees C. Changes in energy metabolism during growth have been reported in many fish species and may reflect the way organisms deal with environmental constraints. We have analyzed enzyme levels (lactate dehydrogenase, LDH: EC 1.1.1.27; and malate dehydrogenase, MDH: EC 1.1.1.37) in four different tissues (white muscle, heart, liver, and brain) from different-sized animals. Both enzymes correlate with body size, increasing the anaerobic potential positively with growth. To our knowledge, this is the first description of scaling effects on hypoxia tolerance and it is interesting to explore the fact that hypoxia survivorship increases due to combining effects of suppressing metabolic rates and increasing anaerobic power as fish grow. PMID- 10817910 TI - The D-galactose-binding lectin of the octocoral Sinularia lochmodes: characterization and possible relationship to the symbiotic dinoflagellates. AB - A D-galactose binding lectin (SLL-2) was isolated from Sinularia lochmodes, an octocoral, by a combination of affinity chromatography on acid-treated agarose and FPLC on Superdex 200. SLL-2 agglutinated rabbit and horse erythrocytes while SLL-1, a minor component, reacted only with rabbit erythrocytes. SLL-2 is a glycoprotein with a molecular mass of 122 kDa and is composed of eight identical subunits (15 kDa). The sequence of the amino terminal region of SLL-2 did not show any apparent homology to the sequences of other animal and plant lectins. D Galactose, N-acetyl-D-galactosamine, lactose, and melibiose were moderate inhibitors to the agglutination of rabbit erythrocytes. In contrast, horse erythrocytes were much more susceptible to agglutination by SLL-2, which was inhibited by sugars and glycoproteins such as D-galactose, N-acetyl-D galactosamine, lactose, melibiose, and porcine stomach mucin. SLL-2 showed considerable tolerance to heating and kept its activity after heating at 80 degrees C for 60 min. In immuno-histochemical studies using an anti-SLL-2 antiserum and protein A gold conjugate, SLL-2 was found to be present in high amounts in the nematocysts. SLL-2 was also detected on the surface of symbiotic dinoflagellate, Symbiodinium sp. cells irrespective whether they were surrounded with or without host cells. These observations suggest the presence of lectin mediated interaction between symbiotic dinoflagellates and S. lochmodes. PMID- 10817911 TI - Cloning and characterization of amphibian cold inducible RNA-binding protein. AB - Gene expression of cold inducible RNA-binding protein (CIRP) was examined in the frog. In Xenopus laevis, expression of CIRP (XCIRP) was observed in both brain and liver at 24 degrees C. Circadian expression of XCIRP was observed in brain. Expression of XCIRP in brain was induced by cold treatment and gradually decreased to the control level at 24 degrees C, but no significant changes were observed in liver. Employing the sequence of murine CIRP, bullfrog (Rana catesbeiana) CIRP gene was cloned. The bullfrog CIRP gene, designated BFCIRP, was 706 bp in length and encoded a putative protein of 164 amino acid residues. The deduced protein contained one consensus sequence of RNA-binding domain (CS-RBD) and a glycine rich domain (GRD). The amino acid sequence of BFCIRP was 78.4% identical to XCIRP. Expression of BFCIRP in brain was stronger in winter than that in summer. These findings suggest that BFCIRP expression in brain may link to hibernation. PMID- 10817912 TI - A protein with inhibitory activity on cell-free translation from cultured mycelia of the edible mushroom Tricholoma lobayense. AB - Cultured mycelia of the edible mushroom Tricholoma lobayense were extracted with cold saline. Proteins were precipitated from the extract by addition of (NH4)2SO4. The precipitate was dissolved and dialyzed before ion exchange chromatography on DEAE-cellulose. Ability to inhibit translation in a rabbit reticulocyte lysate was located in the unadsorbed fraction which was then subjected to affinity chromatography on Affi-gel Blue gel. The strongest activity was again retained by the unadsorbed fraction. Ion exchange chromatography on CM cellulose resulted in fractionation of this fraction into an unadsorbed and two adsorbed peaks. Cell-free translation inhibitory activity was concentrated in the fraction eluted with 100 mM NaCl in 10 mM NH4OAc (pH 5.4). The translation inhibitory protein possessed a molecular weight of 30 kDa as estimated by gel filtration using a fast protein liquid chromatography system and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 10817913 TI - Identification of a myofibril-bound serine proteinase (MBSP) in the skeletal muscle of lizard fish Saurida wanieso which specifically cleaves the arginine site. AB - A myofibril-bound serine proteinase (MBSP) from the skeletal muscle of lizard fish (Saurida wanieso) was purified to homogeneity by a heating treatment followed by a series of column chromatographies on DEAE-Sephacel, Sephacryl S 200, Q-Sepharose, Hydroxyapatite and Benzamidine-Sepharose 6B, and characterized enzymatically. On SDS-poly-acrylamide gel electrophoresis (SDS-PAGE), the purified enzyme showed a band with molecular mass of approximately 29 kDa under reducing conditions, while 60 kDa under non-reducing conditions. The optimum temperature of the enzyme was 50 degrees C using t-butyloxycarbonyl-Phe-Ser-Arg-4 methylcoumaryl-7-amide (Boc-Phe-Ser-Arg-MCA) as a substrate. Substrate specificity analysis both using MCA-substrates and peptides showed that MBSP specifically cleaved at the carboxyl side of the arginine residue. Inhibitor susceptibility analysis revealed that MBSP was inhibited effectively by Pefabloc SC, soybean trypsin inhibitor (STI) and aprotinin, indicating the characteristic of a serine proteinase. When myofibril was incubated with the enzyme, it optically degraded myosin heavy chain at 55-60 degrees C, while alpha-actinin and actin were not at all hydrolyzed as detected by immunoblotting. The N-terminal amino acid sequence of MBSP was partially determined as IVGGAEXVPY- and was very homologous to other serine proteases. PMID- 10817914 TI - Waxes and lipids associated with the external waxy structures of nymphs and pupae of the giant whitefly, Aleurodicus dugesii. AB - The nymphs and pupae of the giant whitefly, Aleurodicus dugesii, produce large quantities of external lipids, both as waxy particles and as waxy filaments. The nymphs and pupae extrude filaments from two dorsal rows of five pores each. Filaments can attain lengths of 5-8 cm. The external lipids of nymphs and pupae consist largely of long-chain aldehydes, alcohols, acetate esters and wax esters. Hydrocarbons are minor components. Soon after hatching, the nymph produced an unidentified waxy fringe extruded laterally from its margin. After molting to the second instar, long, hollow, waxy filaments were produced by the immature stages. The major lipid class associated with the filaments was saturated wax esters (89%), mainly C44, C46 and C60. Associated with formation of the filaments were waxy particles in the shape of curls, which peeled off of the extruding filaments. Similar but more tubular-shaped curls were also produced by numerous lateral pores so that, eventually, the curls completely camouflaged the nymph. The major lipid class of the curls was wax esters (50%), mainly C44 and C46. The cuticular surface lipids of the nymphs were mainly long-chain aldehydes (43%) and wax esters (27%). Unsaturated fatty acid moieties constituted 2 and 19% of the wax esters of curls and nymph cuticular surface lipids, respectively. The major lipid classes of pupae and of their palisade were long-chain aldehydes and alcohols. No unsaturated wax esters were detected in the filaments, but 30% of pupal and 21% of palisade surface wax esters were unsaturated in their fatty acid moieties, 16:1, 18:1 and 20:1. PMID- 10817915 TI - A collectin-like protein from tunicates. AB - Collectins are a sub-family of C-type lectins from mammals and birds that are characterized by their collagen-like domains. The mammalian collectin, mannose binding lectin, has attracted considerable interest because it can activate complement components via a lectin-mediated complement pathway that is independent of immunoglobulins. In this study, we have identified a calcium dependent lectin from the invertebrate (tunicate), Styela plicata, that bears substantial similarities to mammalian collectins. The tunicate lectin, which was isolated by carbohydrate affinity chromatography, has a reduced apparent molecular mass of 43 kDa. The 43 kDa reduced polypeptide appeared as dimers, trimers and hexamers when analyzed by non-reducing and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while gel filtration suggested that the native form of the protein was a nonamer. Amino acid sequence and amino acid composition analysis revealed obvious similarities between the tunicate lectin and mammalian collectins, notably the inclusion of a collagenous domain and a short, cysteine bearing N-terminal domain. The identification of a collectin-like protein in an invertebrate such as S. plicata, which does not express immunoglobulin, indicates that lectin-mediated complement pathways may predate the origin of antibodies. PMID- 10817917 TI - Preface PMID- 10817918 TI - Glycosaminoglycans co-administration enhance insulin-like growth factor-I neuroprotective and neuroregenerative activity in traumatic and genetic models of motor neuron disease: a review. AB - In this report it is shown how glycosaminoglycans and insulin-like growth factor I (IGF-I) promote muscle reinnervation and prevent motor neuron death in experimental models of motor neuron disease. Such effect appears to be mediated by insulin-like growth factor-1. The glycosaminoglycan moiety of proteoglycans is a constituent of the basal lamina active on nerve regeneration by means of the interaction with laminin and with several growth factors. We have previously shown that supplementation by means of subcutaneous injections of glycosaminoglycans affects neuronal degeneration and regeneration. In this study we report that following neonatal lesion of the rat sciatic nerve, glycosaminoglycan treatment promoted extensor digitorum longus muscle reinnervation with consequent improvement of muscle morphology. In saline-treated rats, reinnervation was only partial and there was a marked muscle fibre atrophy, whereas, glycosaminoglycan treatment of lesioned rats increased IGF-I mRNA and protein in the reinnervated muscle, and IGF-I and insulin-like growth factor binding protein-3 plasma levels. Similarly, treatment of lesioned rats with IGF-I promoted muscle reinnervation, and prevented muscle fibre atrophy, higher levels of IGF-I in the reinnervated muscle, of IGF-I, and insulin-like growth factor binding proteins in plasma. In the wobbler mouse IGF-I and glycosaminoglycans alone promote only a partial motor neuron survival and the preservation of forelimb function decays after 3 weeks of treatment. However when glycosaminoglycans and insulin-like growth factor are administered together the motor neuron disease in the wobbler mouse is halted and there is no more loss of motor neurons. PMID- 10817919 TI - Neuroprotective effects of estrogens: potential mechanisms of action. AB - Epidemiological studies associate post-menopausal estrogen use with a reduction in risk of Alzheimer's disease, a reduction in risk of Parkinson's disease, and death from stroke. The neuroprotective efficacy of estrogens have been well described and may contribute to these clinical effects. Estrogen-mediated neuroprotection has been described in several neuronal culture model systems with toxicities including serum-deprivation, beta-amyloid-induced toxicity, excitotoxicity, and oxidative stress. In animal models, estrogens have been shown to attenuate neuronal death in rodent models of cerebral ischemia, traumatic injury, and Parkinson's disease. Although estrogens are known to exert several direct effects on neurons, the cellular mechanisms behind the neuroprotective efficacy of the steroid are only beginning to be elucidated. In this review, we summarize the data supporting a neuroprotective role for estrogens in both culture and animal models and discuss neuronal effects of estrogens that may contribute to the neuroprotective effects. These effects include activation of the nuclear estrogen receptor, altered expression of bcl-2 and related proteins, activation of the mitogen activated kinase pathway, activation of cAMP signal transduction pathways, modulation of intracellular calcium homeostasis, and direct antioxidant activity. PMID- 10817920 TI - Nutritional antioxidants as antidegenerative agents. AB - In this study, primary cultures of cerebellar granule neurons were prepared from eight-day-old Wistar rats, and maintained in an appropriate medium containing a high (25 mM) concentration of KCl. All experiments were performed with fully differentiated neurons (eight days). To induce apoptosis, culture medium was replaced with a serum-free medium (containing 5 mM KCl) eight days after plating. In another series of experiments, apoptosis was induced by application of glutamate (50 microM) to the cell cultures. Apoptosis was measured by flow cytometry, the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP fluorescein nick end-labeling) method, and by the classical method of DNA fragmentation. Since there is evidence that an increased formation of reactive oxygen species (ROS) is involved in the apoptosis induced by both low K(+) concentrations and glutamate, a series of natural antioxidants and a red wine lyophilized extract (which is rich in antioxidant compounds) were tested in our experimental model. It was found that ascorbic acid (30 microM) and a red wine lyophilized extract (5 microgram/ml) were capable of blocking the apoptotic process. Addition of the following natural antioxidants did not have any protective effect on apoptosis induced by low K(+) concentrations: trans- and cis resveratrol (5-200 microM), alpha-tocopherol (100-200 microM), reduced glutathione (100-400 microM), 3-hydroxytirosol (25-100 microM), epicatechin (25 100 microM), or quercetin (25-50 miroM). It is concluded that only a limited number of natural antioxidants are provided with antiapoptotic activity in cultured cerebellar granule neurons. This effect is probably exerted by reducing ROS formation, and by blocking caspase-3 activity. PMID- 10817921 TI - Neurobehavioral aspects of antioxidants in aging. AB - Both aging and age-associated neurodegenerative diseases are associated with various degrees of behavioral impairments, and among the prime candidates responsible for producing the neuronal changes mediating these behavioral deficits appear to be free radicals and the oxidative stress they generate. Therefore, there have been a number of studies which have examined the putative positive benefits of antioxidants in altering, reversing, or forestalling these neuronal/behavioral decrements, with varying degrees of success. Additional experiments have examined the effects of diets rich in fruits and vegetables or herbal extracts in reducing certain types of cancer and cardiovascular diseases, and evidence emerging from such experiments suggests that these kinds of dietary modifications may be beneficial in altering neuronal/behavioral deficits in aging, as well. These kinds of diets are particularly rich in antioxidants such as vitamins A, C, E, and bioflavonoids (such as flavones, tannins, and anthocyanins), and thus, there may be synergistic effects among them. The present paper will review studies concerning the influence of dietary and synthetic antioxidants on normal, pathological age-related, and reactive oxygen species induced behavioral changes in human and animal subjects. The antioxidants reviewed are vitamin E, alpha-lipoic acid, and the phytochemicals contained in herbals, fruits and vegetables. PMID- 10817922 TI - Essential fatty acids and the brain: possible health implications. AB - Linoleic and alpha-linolenic acid are essential for normal cellular function, and act as precursors for the synthesis of longer chained polyunsaturated fatty acids (PUFAs) such as arachidonic (AA), eicosapentaenoic (EPA) and docosahexaenoic acids (DHA), which have been shown to partake in numerous cellular functions affecting membrane fluidity, membrane enzyme activities and eicosanoid synthesis. The brain is particularly rich in PUFAs such as DHA, and changes in tissue membrane composition of these PUFAs reflect that of the dietary source. The decline in structural and functional integrity of this tissue appears to correlate with loss in membrane DHA concentrations. Arachidonic acid, also predominant in this tissue, is a major precursor for the synthesis of eicosanoids, that serve as intracellular or extracellular signals. With aging comes a likely increase in reactive oxygen species and hence a concomitant decline in membrane PUFA concentrations, and with it, cognitive impairment. Neurodegenerative disorders such as Parkinson's and Alzheimer's disease also appear to exhibit membrane loss of PUFAs. Thus it may be that an optimal diet with a balance of n-6 and n-3 fatty acids may help to delay their onset or reduce the insult to brain functions which these diseases elicit. PMID- 10817923 TI - Dietary vitamin E and T cell-mediated function in the elderly: effectiveness and mechanism of action. AB - One of the most dramatic and consequence-bearing age-related phenomena is the decline of the immune function with old age. Age-related T cell-mediated immunity dysfunction has been implicated in the etiology of many of the chronic degenerative diseases of the elderly, including arthritis, cancer, autoimmune diseases and increased susceptibility to infectious diseases. T cells from aged individuals are impaired in their response to mitogens and in their cytokine production. In recent years, several studies have emphasized the importance of intracellular anti-oxidant levels for preserving the immune function. Recent progress in understanding the mechanisms of action of anti-oxidants on cellular metabolism, have shown that anti-oxidants may modulate signal transduction and gene expression in immune cells. Vitamin E is widely recognized as a major lipid soluble chain-breaking anti-oxidant in the biological membrane, where it scavenges free radicals, inhibiting the initiation and chain propagation of lipid peroxidation and protecting cellular structures against oxidative stress damage. Experimental studies have provided evidences for a role of vitamin E in protecting the immune system of elderly subjects. This article reviews the studies concerning the effect of both vitamin E deficiency and supplementation on T cell-mediated immune function in aging. Following a chronological pathway, the present article will also discuss the knowledge regarding the underlying mechanism of action of vitamin E. PMID- 10817924 TI - In vivo microdialysis studies of age-related alterations in potassium-evoked overflow of dopamine in the dorsal striatum of Fischer 344 rats. AB - Intracerebral microdialysis was used to measure basal levels and potassium (K(+)) stimulated overflow of dopamine (DA), homovanillic acid (HVA) and dihydroxyphenylacetic acid (DOPAC), in the dorsal striatum of young (6 months) and aged (24 months) Fischer 344 (F344) rats. Basal levels of HVA were lower in aged rats whereas basal DA and DOPAC did not differ significantly between the two groups. The administration of three low to moderate doses of K(+) (10, 25, and 50 mM) through the microdialysis probe for one collection period revealed differences between the two age groups of F344 rats. DA overflow increased in a dose-dependent manner in the young but not aged rats. Extracellular levels of DOPAC and HVA decreased during the K(+) stimulation and there was a significant difference in the changes in HVA produced by K(+) stimulation in the young vs aged animals. These data support the hypothesis that low to moderate doses of K(+) may be necessary to demonstrate age-related differences in K(+)-evoked DA overflow, since previous microdialysis studies using higher doses have not reported age-related differences in DA overflow. PMID- 10817925 TI - Oxidative damage in Alzheimer's disease: the metabolic dimension. AB - Cell bodies of neurons at risk of death in Alzheimer disease have increased lipid peroxidation, nitration, free carbonyls, and nucleic acid oxidation. These oxidative changes are uniform among neurons and are seen whether or not the neurons display neurofibrillary tangles and, in fact, are acutally reduced in the latter case. In consideration of this localization of damage, in this review, we provide a summary of recent work demonstrating some key abnormalities that may initiate and promote neuronal oxidatave damage. PMID- 10817927 TI - Activation of oncogenic pathways in degenerating neurons in Alzheimer disease. AB - A number of recent findings have highlighted the similarities between neurogenesis during development and neurodegeneration during Alzheimer disease. In fact, neuronal populations that are known to degenerate in Alzheimer disease exhibit phenotypic changes characteristic of cells re-entering the cell division cycle. In this study, we extended these findings by investigating components of the cell cycle, known to trigger progression through G1 through activation of signal transduction cascades. Specifically, we found that proteins implicated in G1 transition, namely Cdc42/Rac, are upregulated in select neuronal populations in cases of Alzheimer disease in comparison to age-matched controls. Importantly, Cdc42/Rac shows considerable overlap with early cytoskeletal abnormalities suggesting that these changes are an extremely proximal event in the pathogenesis of the disease. Given the functional role of Cdc42/Rac in various cellular processes known to be perturbed in Alzheimer disease, namely cytoskeletal organization, oxidative balance, and oncogenic signaling, it is likely that increased neuronal Cdc42/Rac is highly significant in relation to the pathogenic process and contributes to neuronal degeneration. In fact, these findings suggest that Alzheimer disease is an oncogenic process. PMID- 10817926 TI - Induction of cytokines in glial cells surrounding cortical beta-amyloid plaques in transgenic Tg2576 mice with Alzheimer pathology. AB - beta-Amyloid plaque deposition observed in brains from Alzheimer patients, might function as immune stimulus for glial/macrophages activation, which is supported by observations of activated microglia expressing interleukin (IL)-1beta and elevated IL-6 immunoreactivity in close proximity to amyloid plaques. To elucidate the mechanisms involved in beta-amyloid-mediated inflammation, transgenic mice (Tg2576) expressing high levels of the Swedish double mutation of human amyloid precursor protein and progressively developing typical beta-amyloid plaques in cortical brain regions including gliosis and astrocytosis, were examined for the expression pattern of a number of cytokines. Using ribonuclease protection assay, interleukin (IL)-1alpha,-beta, IL-1 receptor antagonist, IL-6, IL-10, IL-12, IL-18, interferon-gamma, and macrophage migration inhibitory factor (MIF) mRNA were not induced in a number of cortical areas of Tg2576 mice regardless of the postnatal ages studied ranging between 2 and 13 months. Using immunocytochemistry for IL-1alpha,beta, IL-6, tumor necrosis factor (TNF)-alpha, and macrophage chemotactic protein (MCP)-1, only IL-1beta was found to be induced in reactive astrocytes surrounding beta-amyloid deposits detected in 14-month-old Tg2576 mice. Using non-radioactive in situ hybridization glial fibrillary acidic protein (GFAP) mRNA was detected to be expressed by reactive astrocytes in close proximity to beta-amyloid plaques. The local immune response detected around cortical beta-amyloid deposits in transgenic Tg2576 mouse brain is seemingly different to that observed in brains from Alzheimer patients but may represent an initial event of chronic neuroinflammation at later stages of the disease. PMID- 10817928 TI - STAT signalling in the mature and aging brain. AB - Activation of the Janus kinases (JAK) and signal transducers and activator of transcription (STAT) proteins in response to specific cytokines and growth factors has been investigated primarily in cells of non-neuronal origin. More recently, the JAKs and the STATs have also been found to be active in the developing and mature brain, providing evidence for important roles played by these molecules in the control of neuronal proliferation, survival and differentiation. Nothing, however, is known about their occurrence and role(s) in the aged brain. We, therefore, investigated the presence of Stat3 and Stat1 in aged-rat brain, and have found that the Stat3 protein was markedly down regulated with respect to adult tissue, while Stat1 remained invariant. We also investigated the potential role of some growth factors in the activation of the JAK/STAT in mature neurons, exposing primary neuronal cells to ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Besides CNTF, which is known to recruit Stat3, we found that Stat3 was also tyrosine phosphorylated by bFGF. These data are indicative of an important role of Stat3 and Stat1 in regulating the physiological status of mature neurons. PMID- 10817929 TI - Contribution of NF-kappaB and p53 in the glutamate-induced apoptosis. AB - Exposure of primary cultures of cerebellar granule cells for 15 min to micromolar concentrations of glutamate results in cell death of both necrotic and apoptotic types. Among the intracellular events triggered by glutamate, we identified two transcriptional factors: the p50 member of the NF-kappaB family and the tumor suppressor phosphoprotein p53. Pretreatment of the cultures with aspirin, which inhibits NF-kappaB activation, or with specific p53 antisense oligonucleotide, which inhibits p53 transcription, resulted in a complete prevention of glutamate induced p53 induction and apoptosis. These findings suggest the existence of a transcriptional program activated by glutamate receptor stimulation in which p50 and p53 play a relevant role. Then, we studied the expression of two p53 downstream genes that could participate in the glutamate-induced pro-apoptotic pathway: p21, which codes for an inhibitor of different cyclin dependent kinases, and MSH2, which codes for a protein involved in the recognition and repair of DNA mismatches. We found that primary cerebellar neurons expressed p21 and MSH2 at very low levels in basal conditions. However, very soon after a brief exposure of the cells to glutamate, the expression of both proteins was dramatically enhanced.On these bases, we propose NF-kappaB, p53, p21 and MSH2 as relevant contributors of the glutamate-induced pro-apoptotic pathway. Understanding this cascade of nuclear events may unravel specific targets for pharmacological intervention for those neurological diseases in which excitatory amino acid induced apoptosis plays a relevant role. PMID- 10817930 TI - NGF-mediated alteration of NF-kappaB binding activity after partial immunolesions to rat cholinergic basal forebrain neurons. AB - There are age-associated cognitive and cholinergic deficits in the neurotrophin dependent cholinergic basal forebrain neurons (CBFNs). There are also increases in the activity of the transcription factor NF-kappaB in the aged rodent brain that may reflect chronic enhancement of stress response signaling. We used partial immunolesions (PIL) to CBFN to examine the role of endogenous NGF on choline acetyltransferase (ChAT) activity and NGF-mediated NF-kappaB alteration after cholinergic deafferentation. We injected 192 IgG-saporin, an immunotoxin selectively taken up by neurotrophin receptor p75(NTR)-bearing neurons, into lateral ventricles, followed by infusions of anti-NGF to assess NF-kappaB, ChAT and NGF responses to PIL after anti-NGF infusion. Treatment with anti-NGF decreased ChAT activity by 17-34% in the cortex, hippocampus, and olfactory bulb and PIL decreased ChAT activity by 47-73%. Changes in AChE activity levels paralleled those observed for ChAT after PIL. NGF protein levels in the olfactory bulb, but not the cortex or hippocampus, increased significantly after PIL treatment. Infusion of anti-NGF abolished the PIL-induced eight-fold NGF increase in CNS. NF-kappaB binding activity to the IgG-kappaB and ChAT specific NF-kappaB consensus sequences, increased in the cortex but not hippocampus after PIL followed by anti-NGF infusion. It is likely that immunolesion-induced changes in ambient NGF levels may perturb NF-kappaB activity. PMID- 10817931 TI - Nerve growth factor brain concentration and stress: changes depend on type of stressor and age. AB - In the relationship between the hippocampus and the hypothalamo-pituitary adrenocortical axis, trophic and tropic actions of nerve growth factor are involved in parallel with those on the cholinergic nuclei of the basal forebrain. Here, we report the changes produced by stress activation of the hypothalamo pituitary-adrenocortical axis on hippocampal and basal forebrain nerve growth factor concentrations in 3-month-old male Wistar rats. The stressors used were: restraint; cold exposure; foot-shock; and rotatory platform. Restraint stress tended to reduce nerve growth factor in the hippocampus and reduced it significantly in the basal forebrain. Nerve growth factor levels in the hippocampus were not modified by cold exposure. However, a single unrepeated exposure significantly increased nerve growth factor in the basal forebrain. Both acute and chronic foot-shock reduced nerve growth factor in the hippocampus, leaving the levels in the basal forebrain unmodified. Acute but not chronic rotatory platform reduced nerve growth factor in the hippocampus, while showing a tendency, more pronounced after chronic application, toward an increase in the basal forebrain. Since with aging both activity of the hypothalamus-pituitary adrenal axis and nerve growth factor trophic and tropic functions change, we studied the effect of restraint and cold stress in the 24-month-old male rat. The variations in nerve growth factor concentrations in the basal forebrain following stress activation are no longer present in the aged rat. The picture that emerges is indicative of a complex relationship between stress and nerve growth factor which is influenced by the kind of stressor and by age. Lack of uniformity in the effects produced by different stressors might reside in different qualitative and/or quantitative degree of involvement of neurotransmitters and/or neurohormones for each of them. PMID- 10817944 TI - Joint contact mechanics in the early stages of osteoarthritis. AB - Joint degeneration in the early stages of osteoarthritis (OA) may be reflected in changes in structural and material properties in articular cartilage. The aim of the present study was to simulate numerically the contact area and stress distribution in normal and "diseased" cartilage layers for dynamic loading. The initial stages of osteoarthritis were simulated based on an experimental model: the anterior cruciate ligament-transected cat knee. In this model, cartilage layers become thicker, softer, and more permeable than the corresponding healthy cartilage layers within weeks of intervention. In our numerical simulations, the diseased cartilage was modelled by changing the thickness, permeability, shear modulus, and Poisson's ratio of the cartilage in accordance with observations in this experimental model of osteoarthritis. The theoretical model of normal and diseased articular cartilage was based on a biphasic representation of cartilage, and the joint was assumed to be axi-symmetric. It was found that, for a given loading condition, the contact areas increase and peak stresses decrease in the diseased compared to the normal joint. According to our simulations, areas of normal joint contact become unloaded and areas of little or no contact become overloaded in the early stages of osteoarthritis compared to the situation in normal joints. Based on these results, we speculate that OA may be initiated following ACL transection because of an overloading of specific regions of the joint, either because of the altered contact mechanics or the disrupted joint stability, despite a general decrease in the contact pressure. PMID- 10817932 TI - Intracellular mechanisms mediating the neuronal death and astrogliosis induced by the prion protein fragment 106-126. AB - Prion encephalopathies include fatal diseases of the central nervous system of men and animals characterized by nerve cell loss, glial proliferation and deposition of amyloid fibrils into the brain. During these diseases a cellular glycoprotein (the prion protein, PrP(C)) is converted, through a not yet completely clear mechanism, in an altered isoform (the prion scrapie, PrP(Sc)) that accumulates within the brain tissue by virtue of its resistance to the intracellular catabolism. PrP(Sc) is believed to be responsible for the neuronal loss that is observed in the prion disease. The PrP 106-126, a synthetic peptide that has been obtained from the amyloidogenic portion of the prion protein, represents a suitable model for studying the pathogenic role of the PrP(Sc), retaining, in vitro, some characteristics of the entire protein, such as the capability to aggregate in fibrils, and the neurotoxicity. In this work we present the results we have recently obtained regarding the action of the PrP 106 126 in different cellular models. We report that the PrP 106-126 induces proliferation of cortical astrocytes, as well as degeneration of primary cultures of cortical neurons or of neuroectodermal stable cell lines (GH(3) cells). In particular, these two opposite effects are mediated by the same attitude of the peptide to interact with the L-type calcium channels: in the astrocytes, the activity of these channels seems to be activated by PrP 106-126, while, in the cortical neurons and in the GH(3) cells, the same treatment causes a blockade of these channels causing a toxic effect. PMID- 10817945 TI - Hemodynamics analyses of arterial expansions with implications to thrombosis and restenosis. AB - It is assumed that critical hemodynamic factors play an important role in the onset, localization and degree of post-operative complications, for example, thrombosis and restenosis. Of special interest are sudden expansion flows, which may occur in straight artery segments such as the common carotid after endarterectomy or end-to-end anastomoses. Sudden expansion geometries are possible origins of early post-operative emboli and significant myointimal hyperplasia resulting in early or late complications. Transient laminar axisymmetric and fully three-dimensional blood flows were simulated employing a validated finite volume code in conjunction with a Runge-Kutta particle tracking technique. Disturbed flow indicators, which may predict the onset of thrombosis and/or restenosis, were identified and employed to evaluate 90 degrees -step and smooth expansion geometries. Smooth expansion geometries have weaker disturbed flow features than step expansion geometries. Specifically, the regions near the expansion wall and the reattachment point are susceptible to both atherosclerotic lesion and thrombi formations as indicated by non-uniform hemodynamic indicators such as near-zero wall shear stress and elevated wall shear stress gradients as well as blood particle accumulation and deposition. A new parameter, the wall shear stress angle deviation (WSSAD) has been introduced, which indicates areas of abnormal endothelial cell morphology and particle wall deposition. In turn, regions of low wall shear stress and high wall shear stress gradients are recognized as susceptible sites for arterial diseases. Thus, it is interesting to note that high WSSAD surface areas cover low wall shear stress, high wall shear stress gradient locations as well as high wall particle deposition.A gradual change in step expansion geometry provides better results in terms of WSSAD values and hence potentially reducing atherosclerosis as well as thrombi formation. PMID- 10817946 TI - Interactions of pulsatile upstream forcing with flow-induced oscillations of a collapsed tube: mode-locking. AB - Interest in flow through collapsible tubes derives from their prevalence in the human body. In the circulation at least, the driving pressure for flow is pulsatile, and it is accordingly appropriate to investigate the response of a collapsible tube to a pulsatile head. A servo-controlled hydraulically powered piston pump was used to add pulsation to the head propelling aqueous flow through a silicone rubber tube. The pump was commanded to execute a sinusoidal variation of piston position at various frequencies on either side of the natural frequency of an operating point where slow (3.75 Hz) repetitive self-excited oscillations in the absence of such forcing could be guaranteed. The resulting time series were characterised by the ratio of the number of collapse cycles to the number of forcing cycles in one overall interaction waveform period. Nonlinear interaction rather than linear superposition was dominant. Many different mode-locked interactions were recorded, and are here systematised in Farey series and plots which show the extent of entrainment. A few of the time series were aperiodic over the time scales investigated, suggesting possible chaos. PMID- 10817947 TI - ECG data compression using optimal non-orthogonal wavelet transform. AB - This paper introduces an effective technique for the compression of electrocardiogram (ECG) signals. The technique is based on a new class of non orthogonal discrete wavelet transform (DWT). The performance of ECG compression algorithm is measured by its ability to minimize distortion while retaining all clinically significant features of the signal. The percent root-mean square difference (PRD) is used as an accepted standard for measuring the signal distortion. However, there is no standard for measuring the clinically significant features retained after signal reconstruction. The coefficients of the DWT are calculated such that the square of the difference between the original signal and the reconstructed one is minimum in least mean square sense. The resulting transforms deal with signals of arbitrary lengths; that means the signal length is not restricted to be a multiple of power of 2. Numerical results comparing the performance of the constructed non-orthogonal transform with that of W-transform and Daubechies D(4) orthogonal transform are given. These results show that, independent of signal length, the decomposition of the signal up to the fourth level is sufficient for getting minimum PRD. In addition, the proposed technique yields the lowest PRD compared to the other two algorithms and for a compression ratio less than 10 the optimal transform can be obtained for only one ECG period. However, for a higher compression ratio the PRD is smaller for long signals. PMID- 10817948 TI - Electropalatography and the Linguagraph system. AB - This paper describes the technique of electropalatography and the development of Linguagraph, which is a user-friendly, clinical instrument, for measurement of tongue/palate contact, during speech. Linguagraph allows objective assessment of tongue function; appropriate targeting of therapy is therefore possible. Visual feedback is also provided, for therapy, and an objective measurement of outcome is easily obtained. Linguagraph was used, for both therapy and assessment, in a clinical trial. Technical aspects of Linguagraph and of the trial results are presented here. These suggest that the instrument will prove useful in the assessment and management of many speech disorders. Full clinical details of the trial are reported elsewhere. PMID- 10817949 TI - Adaptive Morse code communication system for severely disabled individuals. AB - Morse code with an easy-to-operate, single switch input system has been shown to be an excellent communication adaptive device. Because maintaining a stable typing rate is not easy for the disabled, the automatic recognition of Morse code is difficult. Therefore, a suitable adaptive automatic recognition method is needed. This paper presents the application of a Least-Mean-Square algorithm to adaptive Morse code recognition for persons with impaired hand coordination and dexterity. Four processes are involved in this adaptive Morse code recognition method: space recognition, tone recognition, adaptive processing, and character recognition. Statistical analyses demonstrated that the proposed method results in a better recognition rate for the participants tested in comparison to other methods from the literature. PMID- 10817950 TI - Design and testing of a genetic algorithm neural network in the assessment of gait patterns. AB - It is important to be able to quantify changes in gait pattern accurately in order to understand the clinical implications of surgery or rehabilitation. Although supervised feed-forward backpropagation neural networks are very efficient in many pattern-recognition tasks, the genetic algorithm neural network (GANN), which can search in some appropriate space, has not been used previously for gait-pattern recognition. This study discusses how to use the GANN approach in gait-pattern recognition, and evaluates the complexity and training strategy of the particular classification problem. Both the GANN and a traditional artificial neural network (ANN) were used to classify the gait patterns of patients with ankle arthrodesis and normal subjects. The GANN model was able to classify subjects with recognition rates of up to 98.7%. In contrast, the ANN trained by using all possible predictor variables was only able to classify the subjects with recognition rates of 89.7%. It is suggested that the GANN model is more suitable to exploit the patient's gait pattern. The value of the neuron output can be used as an index of the difference from normal. By this means, all pathological gait patterns may be presented quantitatively. PMID- 10817951 TI - Exonic deletion mutations of the Parkin gene among sporadic patients with Parkinson's disease. AB - Exonic deletions of the Parkin gene are common in the autosomal recessive form of juvenile parkinsonism. Here we report Parkin gene mutations among apparently sporadic Parkinson's disease (PD) patients. We screened 200 patients with PD (103 women and 97 men). The age of onset was 54.2+/-10.3years (mean+/-S.D.).Four out of the 200 patients had homozygous exonic deletions in the Parkin gene. The clinical features of these four patients were essentially the same as those of idiopathic PD. The age of onset was consistently younger (33, 38, 47 and 48years, respectively). On medication, all of them were at Hoehn and Yahr stage II or III even after 12-16years from the onset of the disease.Thus 2% of apparently sporadic PD patients in Japan have homozygous Parkin gene mutations. This positive rate was 6.3% among the patients with the age of onset below 50. Our study suggests that the prevalence of the carrier state of Parkin gene may be more than that we expected. Our study warrants further studies on Parkin gene mutations in apparently sporadic PD patients. PMID- 10817952 TI - Unilateral pallidotomy for Parkinson's disease: speech, motor, and neuropsychological outcome measurements. AB - We studied the effects of unilateral medial pallidotomy in the first 57 consecutive Parkinson's disease (PD) patients undergoing this MRI/electrophysiologically guided procedure at our institution, obtaining qualitative and quantitative measures of speech, motor function, activities of daily living, and neuropsychological functioning at 3 and 12months post operatively. Speech intelligibility was typically preserved, declining mildly in one-third of patients post-operatively. Pallidotomy significantly improved motor function similarly in patients >/=65 or <65years (n=20). Cognitive abilities generally remained stable following surgery; however, performance on measures of letter fluency and semantic fluency declined in patients with left pallidotomies. We conclude that in our series mild deterioration in speech may occur with unilateral pallidotomy in patients otherwise responsive to this treatment. PMID- 10817953 TI - The epidemiology of progressive supranuclear palsy (Steele-Richardson-Olszewski syndrome). AB - Progressive supranuclear palsy (PSP, also known as Steele-Richardson-Olszewski syndrome) is relatively uncommon. Studies to date have found a crude population prevalence for PSP of between 1.39 and 4.9 cases per 100,000. An increased awareness of the condition, together with recently formulated clinical diagnostic criteria, should facilitate future descriptive epidemiological studies.Possible aetiological influences both genetic (tau polymorphisms) and environmental (herbal tea and tropical fruit consumption in the French West Indies) have recently been postulated. Future studies into both disease prevalence and aetiology would benefit from a multi-centre approach. PMID- 10817954 TI - Gender distribution in surgery for Parkinson's disease. AB - Parkinson's disease (PD) affects both women and men. The surgical treatment of this disease has experienced a worldwide increase since the mid-eighties. In order to document eventual differences in gender distribution of patients undergoing various stereotactic surgical procedures for PD, we reviewed scientific papers published during the last 14years.A literature search provided 145 clinical papers, published between January 1985 and February 1999, and dealing with pallidotomy, thalamotomy and deep brain stimulation procedures. These papers were scrutinised with respect to redundancies or other overlap of reported patients. The resulting numbers of patients were compiled according to gender, to surgical procedure, and to geographic area of origin of the publishing centers.In one third of the reviewed publications the gender of the patients was not specified. In the remaining papers, the overall sex distribution of patients who underwent surgery was 35% females and 65% males. These proportions between sexes were relatively consistent regardless of surgical procedure, and regardless of geographic origin of the publications.Male preponderance in patients undergoing surgery for PD cannot be explained by a corresponding difference in gender-prevalence of the disease. The criteria of selection, and patterns of referral, of patients for surgery, as well as the respective attitude of female and male patients toward surgery, may account for the uneven gender distribution in surgical PD patients. PMID- 10817955 TI - Effects of histamine H(3)-ligands on the levodopa-induced turning behavior of hemiparkinsonian rats. AB - Histamine H(3)-receptors act as heteroreceptors on many neurons. The effects of H(3)-ligands (an agonist, R-alpha-methylhistamine and an antagonist, thioperamide) on levodopa-induced turning behavior in a rat model of Parkinson's disease were quite similar to those seen with alpha(2)-adrenoceptor ligands (dexmedetomidine and atipamezole). R-alpha-methylhistamine clearly reduced contralateral turning behavior but the increase of turning behavior after thioperamide was less clear. The lack of effect of H(3)-ligands, in contrast to alpha(2)-ligands, on the amphetamine-induced ipsilateral turning behavior points to different roles or neuronal distribution of these two presynaptic receptors. We propose that in this lesion model, H(3)-receptors modify those pathways participating striatal outflow. PMID- 10817956 TI - Construction of freezing of gait questionnaire for patients with Parkinsonism. AB - Background: Freezing of gait (FOG) is a common, poorly understood, parkinsonian symptom interfering with daily functioning and quality of life. Assessment of FOG is complex because of the episodic nature of this symptom, and the influence of mental and environmental factors on it.Objective: To design a self-reportable reliable questionnaire for FOG.Method: A questionnaire consisting of 16 items regarding gait and falls was administered together with the Unified Parkinson's Disease Rating Scale (UPDRS) to 40 Parkinson's disease (PD) patients (26 males) with a mean age of 72.3+/-9.3 years and mean Hoehn and Yahr (H&Y) stage at "Off" of 2.85+/-0.84. A principal component analysis with Varimax rotation was conducted on the results. Item analyses were performed and reliability computed for an abbreviated FOG questionnaire.Results: Based on these analyses, a short (six item) FOG questionnaire was constructed, which was found to be highly reliable (Cronbach alpha=0.94) for assessment of FOG and with moderate correlation with the activity of daily living (ADL) and motor parts of the UPDRS (0.43 and 0.40, respectively). Moderate correlation was also found with the FOG item at the ADL part of the UPDRS (alpha=0.66 for the "Off" and 0.77 for the "On" state).Conclusion: The FOG questionnaire that was constructed is highly reliable in assessing freezing of gait, unrelated to falls, in patients with PD. PMID- 10817957 TI - Levodopa in plasma correlates with body weight of parkinsonian patients. AB - Background: Antiparkinsonian comedication and/or altered gastrointestinal motility and absorption influence plasma levels of levodopa. Another putative factor may represent body weight. The objective of our study was to examine the relationship between body weight and levodopa in plasma of parkinsonian patients.Methods: We enrolled 26 individuals into this trial on the resorption of levodopa. A standardized protocol was used, which eliminated influencing factors, such as comedication, activity, food and estimated levodopa plasma levels at fixed timepoints.Results: Levodopa amount (Spearman R=-0.48; p=0.013) and maximum concentration (Spearman R=-0.50; p=0.008) in plasma correlated negatively with body weight.Conclusion: Doses of levodopa should be considered in relation to body weight. Thus severe loss of weight may reduce the need for oral levodopa intake by parkinsonian subjects and may represent a putative factor in the occurrence of dyskineasia in these patients unless there is a concomitant reduction in the dose of oral levodopa. PMID- 10817958 TI - Intrafamilial phenotypic variability of Hallervorden-Spatz syndrome in a Tunisian family. AB - We report clinical features, CT-scan and MRI findings of three siblings with Hallervorden-Spatz disease, one of them followed for more than 20years. Patient 1 presented at age 10 with progressive generalized dystonia. Five years later she had violent dystonic spasms with opisthotonos and marked oro-mandibular involvement. Later, dystonia stabilized and examination showed dysarthria, multiple deformities related to dystonic posturing, retinal degeneration and no cognitive impairment. Cerebral CT-scan showed bilateral pallidal lucencies and T2 cerebral MRI showed the "eye of the tiger" sign.Patient 2 developed delusions and hallucinations at age 18 with acute phases of hetero-aggressivity diagnosed as "schizophrenic" in a psychiatric hospital. At age 20, he developed oromandibular dystonia, severe dysarthria and epilepsy. Cerebral MRI showed the same "eye of the tiger" sign.Patient 3 presented at age 37 with isolated psychiatric features similar to those of patient 2, also diagnosed "schizophrenic" in a psychiatric hospital. Neurological examination showed mild postural tremor of the hands and intermittent cervical dystonia. He was stabilized with neuroleptic treatment.This family shows marked intrafamilial variability of age at onset, symptom at onset (with "schizophrenic" features unusually described and leading to misdiagnosis), clinical presentation (almost entirely dystonic in one patient and almost entirely psychiatric in two others) and course of the disease (slowly progressive in one sibling, rapidly deteriorating in the second and stabilization in the third). Classification and nosology of subtypes of HSD are discussed. PMID- 10817959 TI - Cortical activation in self-paced versus externally cued movements: a hypothesis. PMID- 10817960 TI - Psychological functioning in patients with Parkinson's disease. AB - Forty patients with Parkinson's disease (PD) were studied to explore the relationship between aspects of psychological functioning (depression, problems in motoric, cognitive and psychological areas) and physical illness (stage and duration of illness, functional disability, global cognitive status). Depression, motoric, psychological and cognitive problems were found to be significantly greater in patients suffering from severe as compared to moderate disease. Functional disability correlated more strongly with variables assessing psychological functioning. It was concluded that problems of patients with PD were not restricted to the motoric domain but are multi-dimensional in nature, and other factors should be taken into account in the clinical management of the illness. These findings have implications for development of interventions for patients with PD. PMID- 10817961 TI - Role of Oncostatin M in hematopoiesis and liver development. AB - Definitive hematopoietic stem cells (HSCs) first appear in the aorta/gonad/mesonephros (AGM) region and migrate to the fetal liver where they massively produce hematopoietic cells before establishing hematopoiesis in the bone marrow at a perinatal stage. In the AGM region, Oncostatin M (OSM) enhances the development of both hematopoietic and endothelial cells by possibly stimulating their common precursors, so-called hemangioblasts. During development of HSCs in the AGM region, the liver primodium is formed at the foregut and accepts HSCs. While fetal hepatic cells function as hematopoietic microenvironment for expansion of hematopoietic cells during mid to late gestation, they do not possess most of the metabolic functions of adult liver. Along with the expansion of hematopoietic cells in fetal liver, OSM is produced by hematopoietic cells and induces differentiation of fetal hepatic cells, conferring various metabolic activities of adult liver. Matured hepatic cells then lose the ability to support hematopoiesis. Thus, OSM appears to coordinate the development of liver and hematopoiesis in the fetus. PMID- 10817962 TI - Can cord blood cells support the cytokine storm in GvHD? AB - Cord blood has a high number of proliferating hematopoietic progenitors and is therefore used as an alternative source of hematopoietic cells for allogeneic transplantation. In addition there is a wider availability of cord blood and a lower cost of procurement compared to bone marrow. However one of the most interesting immunological benefits of a cord blood transplant that has been proposed is the low severity of Graft versus Host Disease (GvHD). This review aims to address some of the immunological reasons why this may be the case by assessing the role of cord blood cytokines in the cytokine storm of GvHD. PMID- 10817963 TI - STAT family of transcription factors in cytokine-mediated biological responses. AB - STAT (signal transducers and activators of transcription) family, consisting of seven members, is involved in cytokine signal transduction. Biological roles of each STAT family protein have now been elucidated through studies of gene targeted mice. Stat1 knockout mice are defective in interferon-mediated functions. Stat4 and Stat6 knockout mice show defective responses to IL-12 and IL 4, respectively. Analyses of Stat5a and Stat5b knockout mice reveal important roles in prolactin-mediated mammary gland development and growth hormone-mediated induction of sexual dimorphism, respectively. Conditional knockout study of Stat3 demonstrates its critical roles in cytokine-mediated functions in several tissues, including T cells, macrophages, skin, and mammary gland. PMID- 10817964 TI - LIRs/ILTs/MIRs, inhibitory and stimulatory Ig-superfamily receptors expressed in myeloid and lymphoid cells. AB - Cells exhibit a complex network of inhibitory and stimulatory signaling pathways, which interact with each other to maintain an homeostatic balance and modulate cellular responses to external stimuli. During most of the 1980s, a great effort was put into the characterization of stimulatory cell surface receptors for cytokines and growth factors. In the last decade, a large number of inhibitory receptors have been identified and it has become apparent that inhibitory signaling pathways are subject to intricate regulatory mechanisms. Inhibitory and stimulatory signaling pathways work in concert with each other to establish activation thresholds and provide sensitive tuning mechanisms that help control cellular responses. LIRs/ILTs/MIRs are a novel family of inhibitory and stimulatory receptors expressed both in myeloid and lymphoid cells. They contain two or four immunoglobulin-like domains in the extracellular region and their cytoplasmic domains are either very short and without any signaling motifs or are long and contain a variable number of immunoreceptor tyrosine-based inhibition motifs (ITIMs). LIRs within the first group send stimulatory signals by association with the FcR common gamma chain and LIRs within the second group deliver inhibitory signals by association with the protein tyrosine phosphatase SHP-1. This review summarizes our current knowledge on the LIRs, their ligands, and biological functions. PMID- 10817965 TI - The biology of Toll-like receptors. AB - In 1997, a human homologue of the Drosophila Toll protein was described, a protein later to be designated Toll-like receptor 4 (TLR4). Since that time, additional human and murine TLR proteins have been identified. Mammalian TLR proteins appear to represent a conserved family of innate immune recognition receptors. These receptors are coupled to a signaling pathway that is conserved in mammals, insects, and plants, resulting in the activation of genes that mediate innate immune defenses. Numerous studies have now identified a wide variety of chemically-diverse bacterial products that serve as putative ligands for TLR proteins. More recent studies have identified the first endogenous protein ligands for TLR proteins. TLR signaling represents a key feature of innate immune response to pathogen invasion. PMID- 10817966 TI - Regulation of retinoic acid signaling in the embryonic nervous system: a master differentiation factor. AB - This review describes some of the properties of retinoic acid (RA) in its functions as a locally synthesized differentiation factor for the developing nervous system. The emphasis is on the characterization of the metabolic enzymes that synthesize and inactivate RA, and which determine local RA concentrations. These enzymes create regions of autocrine and paracrine RA signaling in the embryo. One mechanism by which RA can act as a differentiation agent is through the induction of growth factors and their receptors. Induction of growth factor receptors in neural progenitor cells can lead to growth factor dependency, and the consequent developmental fate of the cell will depend on the local availability of growth factors. Because RA activates the early events of cell differentiation, which then induce context-specific differentiation programs, RA may be called a master differentiation factor. PMID- 10817967 TI - Cytokines revisited at Hilton head. AB - The Seventh International Cytokine Society Meeting, held in December 1999 at Hilton Head, provided once more an ideal setting for an intense discussion and remodeling of concepts regarding the biology and molecular biology of cytokines and chemokines. Approximately 400 scientists gathered at the beach-side resort, forgoing the beauty of the ocean to listen to their colleagues in the field, in a program ably organized by Jan Vilcek, Scott Durum, Bruce Beutler and Ann Richmond. Here, we attempt to cover some of the highlights on gene regulation, T helper cell differentiation, signal transduction, receptor trafficking, chemokines, apoptosis, disease models, and new cytokines. PMID- 10817968 TI - Status of anti-HIV-1 chemotherapy in Japan. PMID- 10817969 TI - Development of Male Flowers in Zizania aquatica (North American Wild-Rice; Gramineae). AB - We investigated the histology and developmental morphology of flowers of wild rice (Zizania aquatica), a member of grass subfamily Oryzoideae, to determine whether male flowers in this species develop in a manner similar to those in the subfamily Panicoideae, a group that includes many species with unisexual flowers. Zizania has evolved unisexual flowers from hermaphrodite ancestors and is only distantly related to the Panicoideae; the origins of unisexual flowers are independent in the two taxa. Ontogenetic evidence indicates that many species within the subfamily Panicoideae develop male flowers by a process similar to that already described for maize (Zea mays), a panicoid grass. Unisexual male flowers in maize initiate both the stamen (androecium) and the pistil (gynoecium), but the gynoecium aborts early in development through the death of the subepidermal cells. Cell death in gynoecia of maize is known to be controlled by the product of the gene tasselseed2 (ts2), and an orthologue of ts2 has been shown to have the same effect in the sister genus Tripsacum. It seems likely that ts2 orthologues mediate cell death throughout the Panicoideae, but the phylogenetic range of the cell death mechanism is not known. In this study we show that male flowers of Z. aquatica show neither the distinctive pattern of cell death nor the ontogenetic timing of abortion that are characteristic of male flower formation in studied species of Panicoideae. This indicates that these unisexual flowers may be produced by an entirely different mechanism from that employed by the Panicoideae. Either ts2 does not control sex expression in Zizania, or it is deployed at a different time, and possibly in different tissues, with a different histological result. Our results indicate that the independent origins of male flowers in Gramineae apparently do not have a common system of genetic control. PMID- 10817970 TI - Histology of Organogenic and Embryogenic Responses in Cotyledons of Somatic Embryos of Quercus Suber L. AB - In cork oak (Quercus suber L.), recurrent embryogenesis is produced in vitro through autoembryony without exogenous plant growth regulators (PGRs); secondary embryos appear on the embryo axis but seldom on cotyledons. Focusing mainly on the histological origin of neoformations, we investigated the influence of the embryo axis and exogenous PGRs on the embryogenic potential of somatic embryo cotyledons. Isolated cotyledons of somatic embryos became necrotic when cultured on PGR-free medium but gave secondary embryos when cultured on media containing benzyladenine and naphthaleneacetic acid. Cotyledons of cork oak somatic embryos are competent to give embryogenic responses. Isolated cotyledons without a petiole showed a lower percentage of embryogenic response than did those with a petiole. In petioles, somatic embryos arose from inner parenchyma tissues following a multicellular budding pattern. Joined to the embryo axis, cotyledons did not show morphogenic responses when cultured on PGR-free medium but revealed budlike and phylloid formations when cultured on medium with PGRs. The different morphogenic behavior displayed by somatic cotyledons indicates an influence of the embryo axis and indicates a relationship between organogenic and embryogenic regeneration pathways. PMID- 10817971 TI - Haploid Origin of Cork Oak Anther Embryos Detected by Enzyme and RAPD Gene Markers. AB - In vitro-induced cork oak (Quercus suber L.) embryos from anther cultures proved to be of haploid origin both by enzyme and RAPD gene marker analysis. The problem considered was to ascertain if embryo cultures originated either from a single haploid cell, from a microspore, or from multiple haploid cells. Therefore, a heterozygotic gene was searched for in the parent tree. The gene coding for shikimate dehydrogenase (SKDH1) proved to be heterozygous in the parental tree, and subsequently, these allozymes were screened for the embryos induced in anther cultures from the same tree. Only haploid embryos were found, confirming the microspore origin. Different genotypes were not identified inside each anther by isozyme analysis, probably because of selective pressure for one embryo early in development, but both parental SKDH1 alleles were found in the embryos of different anthers. The banding patterns detected by RAPD markers permitted the identification of multiple microspore origins inside each anther. PMID- 10817972 TI - Supercooling Capacity Increases from Sea Level to Tree Line in the Hawaiian Tree Species Metrosideros polymorpha. AB - Population-specific differences in the freezing resistance of Metrosideros polymorpha leaves were studied along an elevational gradient from sea level to tree line (located at ca. 2500 m above sea level) on the east flank of the Mauna Loa volcano in Hawaii. In addition, we also studied 8-yr-old saplings grown in a common garden from seeds collected from the same field populations. Leaves of low elevation field plants exhibited damage at -2 degrees C, before the onset of ice formation, which occurred at -5.7 degrees C. Leaves of high-elevation plants exhibited damage at ca. -8.5 degrees C, concurrent with ice formation in the leaf tissue, which is typical of plants that avoid freezing in their natural environment by supercooling. Nuclear magnetic resonance studies revealed that water molecules of both extra- and intracellular leaf water fractions from high elevation plants had restricted mobility, which is consistent with their low water content and their high levels of osmotically active solutes. Decreased mobility of water molecules may delay ice nucleation and/or ice growth and may therefore enhance the ability of plant tissues to supercool. Leaf traits that correlated with specific differences in supercooling capacity were in part genetically determined and in part environmentally induced. Evidence indicated that lower apoplastic water content and smaller intercellular spaces were associated with the larger supercooling capacity of the plant's foliage at tree line. The irreversible tissue-damage temperature decreased by ca. 7 degrees C from sea level to tree line in leaves of field populations. However, this decrease appears to be only large enough to allow M. polymorpha trees to avoid leaf tissue damage from freezing up to a level of ca. 2500 m elevation, which is also the current tree line location on the east flank of Mauna Loa. The limited freezing resistance of M. polymorpha leaves may be partially responsible for the occurrence of tree line at a relatively low elevation in Hawaii compared with continental tree lines, which can be up to 1500 m higher. If the elevation of tree line is influenced by the inability of M. polymorpha leaves to supercool to lower subzero temperatures, then it will be the first example that freezing damage resulting from limited supercooling capacity can be a factor in tree line formation. PMID- 10817973 TI - Effect of Nutrients on Growth Rate and Carbohydrate Storage in Oxytropis sericea: A Test of the Carbon Accumulation Hypothesis. AB - The pool of storage carbohydrates in plants may be formed in competition with allocation to growth ("true reserves") or, where growth is inhibited by external factors, with no cost to growth ("accumulated reserves"). The latter hypothesis was addressed using Oxytropis sericea, a perennial herb from the Rocky Mountains. To generate variation in growth rate, greenhouse-cultivated plants were assigned to low or high nutrient levels in 2 yr. Age of plants was also varied either by conducting consecutive harvests (1996) or using plants of different initial age (1997). In both years, plants from the high-nutrient group attained larger vegetative biomass than plants in the low-nutrient group and also displayed an overall lower carbon allocation to storage, consistent with the accumulation hypothesis. PMID- 10817974 TI - Touch-Sensitive NADH Oxidase Activity of Pea and Cucumber Tendrils and Soybean Hypocotyl Sections. AB - The cell surface reduced nicotinamide adenine dinucleotide (NADH) oxidase activity of soybean stems and of pea and cucumber tendrils responded to touch with a several-fold increase in activity. The increase in NADH oxidase persisted for 20 min or longer, and further touch stimulation during this period did not alter activity. With soybean sections, the specific activities in response to touch approximated those achieved maximally by auxin. Where the NADH oxidase was fully stimulated by 2,4-d, the NADH oxidase failed to respond further to touch. The findings indicate that the NADH oxidase of the plant cell surface is involved in the growth response to touch and in tendril coiling. PMID- 10817975 TI - Self-Incompatibility and Male Fertilization Success in Phillyrea angustifolia (Oleaceae). AB - Androdioecy is a rare breeding system in which low male frequency is expected in populations because males require a strong increase in their fertility to be maintained by selection. Phillyrea angustifolia L. has previously been reported as possibly functionally androdioecious. However, 1&rcolon;1 sex ratios have been reported and suggest functional dioecy. In this article, we compared both pollen tube growth and siring success of male and hermaphrodite pollen in two single donor pollination experiments. We verified at both pre- and postzygotic levels that hermaphrodites produce functional pollen. Self-incompatibility was also clearly established. However, pollen from hermaphrodites was less efficient than male pollen. The probability of a pollen tube growing through the style was higher for male than for hermaphrodite pollen donors, and males sired twice as many fruits as hermaphrodites. The twofold male advantage in relative fecundity was mainly because of lower pollen fertility of hermaphrodites and possible cross incompatibility among hermaphrodites. PMID- 10817976 TI - Genetic Diversity in Alpine and Foothill Populations of Campanula rotundifolia (Campanulaceae). AB - Climatic constraints on insects in alpine environments have important consequences for the biology of their plant mutualists; in particular, reduced insect diversity and activity in alpine plant populations can result in pollinator-limited seed set and, potentially, in low genetic diversity. However, highly effective pollination by bumblebees in alpine populations can compensate for low visitation rates. In this study we hypothesized that, because of highly effective pollination by bumblebees, alpine populations of Campanula rotundifolia would not experience more frequent cycles of pollinator limitation than low elevation populations and would therefore exhibit comparable levels of genetic variability and inbreeding to those found in foothill populations. Enzyme electrophoresis was used to assess genetic variability at nine putative loci in alpine and foothill populations of C. rotundifolia in Colorado. Genetic variability in C. rotundifolia was found to be comparable to that reported for other long-lived herbaceous perennials. Measures of genetic variability and fixation indices did not differ between high- and low-elevation populations and were consistent with Hardy-Weinberg expectations. Nonsignificant F(ST) values indicated no genetic differentiation among all populations. PMID- 10817977 TI - Allozyme Variation and Genetic Relationships among Species of Cimicifuga (Ranunculaceae) from Korea. AB - Allozyme investigation of the five Cimicifuga taxa in Korea was conducted to assess genetic and clonal diversity within populations and genetic divergence among populations and taxa. Levels of allozyme variation maintained in Korean Cimicifuga taxa were comparable to those for most herbaceous perennials. In general, samples excluding copies of the same multilocus genotype maintained higher levels of genetic diversity than the total samples within populations. Copies of homozygous genotypes at several loci resulting from clonal spread lead to decreased levels of genetic diversity within populations, indicating that clonal reproduction found in Cimicifuga affects population genetic structure. In general, more widely distributed species such as C. dahurica and C. japonica harbored higher levels of allozyme diversity than the other taxa examined. Although two varieties of C. heracleifolia are geographically and reproductively isolated, the genetic and clonal structure of var. bifida seems to resemble var. heracleifolia, indicating that the two varieties may have had a similar evolutionary history. However, the allozyme data strongly indicate that the two morphological types (Groups I and II) of C. simplex should be treated as separate species. PMID- 10817978 TI - A Phylogenetic Study of Tribe Euphorbieae (Euphorbiaceae). AB - A phylogenetic investigation of a monophyletic lineage of spurge plants, tribe Euphorbieae, was conducted to elucidate evolutionary relationships, to clarify biogeographic patterns, and to reexamine the previous classification of Euphorbieae. Cladistic analyses of the 52 morphological characters of 61 species resulted in 2922 equally most parsimonious trees of 193 steps with a consistency index of 0.34. The strict consensus tree indicates genus Anthostema of subtribe Anthosteminae as a likely sister group to all other members of tribe Euphorbieae. The morphological data support a monophyletic origin of subtribe Euphorbiinae, but the subtribes Anthosteminae and Neoquillauminiinae did not form monophyletic groups. Although the previous taxonomic treatments within tribe Euphorbieae have supported the generic status of Pedilanthus, Monadenium, Synadenium, Chamaesyce, and Elaeophorbia, the results of this analysis do not support generic placement of them based on cladistic principles. Recognition of these groups as genera results in Euphorbia becoming a paraphyletic group. One solution to this problem in Euphorbieae is to divide the largest genus Euphorbia into several monophyletic genera and to keep the generic ranks for previously recognized genera. The distribution of basal endemic genera in Euphorbieae showed African and east Gondwanan affinities and strongly indicated that the ancestor of Euphorbieae originated prior to the breakup of Gondwanaland from an old group in Euphorbiaceae. However, some recent African taxa of Euphorbia should be interpreted by transoceanic dispersal from the New World ancestors. PMID- 10817979 TI - Microstructure of Purified Rubber Particles. AB - Purified rubber particles from Hevea brasiliensis (Brazilian rubber tree), Parthenium argentatum (guayule), Ficus elastica (Indian rubber tree), and Euphorbia lactiflua were examined and compared using conventional scanning electron microscopy (SEM), field-emission SEM, cryo-SEM, and transmission electron microscopy (TEM). Rubber particles of all four species were spherical; they varied in size and had a uniform homogeneous material, the rubber core, surrounded by a contiguous monolayer (half-unit) membrane. Frozen-hydrated and/or untreated particles from H. brasiliensis and P. argentatum deformed and fused readily, whereas those from F. elastica and E. lactiflua retained their spherical shapes. These results indicate that the surface components of the H. brasiliensis and P. argentatum particles are more fluid than those of F. elastica or E. lactiflua. When fixed in aldehyde, F. elastica particles retained their spherical exterior shapes but had hollow centers, whereas H. brasiliensis and P. argentatum particles completely collapsed. In aldehyde-osmium tetroxide-fixed material, the rubber core of F. elastica was poorly preserved in some particles in which only a small amount of the rubber core remained adhering to the monolayer membrane, leaving a hollow center. Euphorbia lactiflua particles were well preserved in terms of retaining the rubber core; however, the membrane was not as easily discernible as it was in the other three species. Both H. brasiliensis and P. argentatum were well preserved following fixation; their cores remained filled with rubber, and their monolayer membranes were defined. The addition of potassium permanganate to the fixation-staining regime resulted in higher contrast micrographs and more well defined monolayer membranes. PMID- 10817980 TI - Compressive/Tensile Stresses and Lignified Cells as Resistance Components in Joints between Cladodes of Opuntia laevis (Cactaceae). AB - The Cactaceae are a diverse group of plants with a wide variety of morphologies. Many species of Opuntia have segmented stems in which terminal cladodes may be separated from main-stem cladodes with varying amounts of resistance. From a geometric approach, derivations were used to calculate normal (axial and bending) and shear (transverse force and torque) stresses at joints due to the weight of the cladodes. Normal and shear stresses act perpendicular and parallel to (along) the cross sections of joints, respectively. Normal stress caused by bending was >10 times that of the mean value of any other stress. Analyses were performed to determine the relationship between maximum normal stress and the amount of lignified xylem cells. Such cells had thicker cell walls compared with the various other cells of stem joints that had thin cell walls and that thus would provide the most resistance to normal stresses. An analogy was made between cactus joints and a composite beam with reinforcing rods. In such joints, thin walled parenchyma cells might be analogous to concrete that has little resistance to tensile stress, while the thick-walled, lignified xylem cells would be analogous to reinforcing rods. There were statistically significant relationships between normal stresses (from bending and axial loads) and mean percentage of lignified xylem cells (r=0.73) and between normal stresses and total areas of lignified xylem cells (r=0.65) (more stress, more reinforcing xylem cells). Tensile portions of cactus joints had 23% lignified xylem cells, while compressive portions had only 10% lignified xylem cells in joint areas (more tension, more reinforcing xylem cells). In addition, tensile joint tissues had two to three times more thick-walled, lignified xylem cells in the outer 30% of the radius compared with other joint tissues types (more reinforcing near the surface). To our knowledge, this is the first publication to present mechanical stresses at stem joints of cacti and the first to relate these stresses to characteristics of resisting tissues in the joints of a cactus. PMID- 10817981 TI - Cytohistochemical Analysis of Malus domestica Borkh. Seeds from Shedding and Nonshedding Fruits. AB - A histological, cytochemical, and ultrastructural investigation of seeds of control (nonshedding) and shedding fruits of Malus domestica Borkh. showed differences in the pattern of nutrient distribution. The kind and amount of storage compounds in the two seeds also differed. Protein bodies and starch were abundant in the control seeds and nearly absent in the seeds of shedding fruit. In the latter, there was a great abundance of phenols that exhibited a specific distribution in different tissues of the chalazal region. A cuticle was present on the inner side of the inner integument of the seeds of both control and shedding fruits. This cuticle was interrupted in the chalaza and formed a pathway for nutrient translocation, what we termed the "window." Cell wall changes observed in the chalazal window were probably associated with the different distribution pattern of storage compounds and with the control of nutrient routing. The delayed and variable development of the embryos of seeds in shed fruits may be related to both the altered pattern of storage substances and their translocation to the embryo. PMID- 10817982 TI - A Reconsideration of Hyenia elegans Krausel et Weyland and Hyenia 'complexa' Leclercq: Two Middle Devonian Cladoxylopsids from Western Europe. AB - Two taxa of Middle Devonian Cladoxylopsida from localities in Belgium and Germany are redescribed. A counterpart of one of the missing type specimens of Hyenia elegans Krausel et Weyland from Kirberg, Germany, demonstrates that rather than having simple forked fertile appendages (as apparent from the type description), more complex fertile units were present; these consisted of two daughter segments (resulting from a proximal dichotomy), each of which bore three pairs of elongate sporangia on short recurved stalks proximal to a terminal bifurcation. Specimens from Oe, Belgium, informally referred to as Hyenia 'complexa' by Leclercq, were shown to have a similar fertile organ that lacked the tuft of filiform extensions previously believed to be present. The parallel axes of this plant are now interpreted to be derived from a digitate branch rather than from a horizontal rhizome. Both taxa, H. elegans and H. 'complexa,' therefore demonstrate fertile appendage morphology and other characters that are typical of the Middle Devonian cladoxylopsid Calamophyton primaevum Krausel et Weyland, which has priority. PMID- 10817983 TI - Reconstructing the Pennsylvanian-Age Filicalean Fern Botryopteris tridentata (Felix) Scott. AB - Numerous anatomically preserved fragments of the Middle Pennsylvanian age filicalean fern, Botryopteris tridentata, occur in coal balls collected at the Pittsburgh and Midway Coal Company mine near Baxter Springs, Kansas. Included are the first fertile specimens of the species, evidence of complete vegetative frond architecture, and fronds that are specialized for vegetative propagation. Rhizomes are erect and unbranched, have helical phyllotaxis and short internodes, and typically display an ectophloic solenostele. Fronds are tripinnately compound with lobed pinnules that have open, dichotomous venation. Fertile pinnae or individual pinnules are interspersed among vegetative frond segments and produce sori of annulate sporangia beneath veins on the abaxial pinnule surface. Fertile pinnule lobes are rolled toward the abaxial surface to enclose the sori. Sporangia have a horizontally elongated biseriate annulus located near the short broad stalk and produce tetrahedral-shaped trilete spores with coarse spines. Epiphyllous branches diverge from the stipe or rachis, and some fronds produce only branches. This fern is reconstructed as having short stems. Helically arranged fronds are either pinnately dissected with lobed vegetative pinnules and abaxially rolled fertile pinnules or are specialized for vegetative propagation. The latter functioned as the foliar equivalent of stolons. While some characters of the B. tridentata plant are similar to those of Botryopteris forensis, generitype of the Botryopteridaceae, others are more comparable to those of Psalixochlaena cylindrica, generitype of Psalixochlaenaceae, suggesting the need for reevaluation of systematic relationships among species of the Botryopteridaceae and Psalixochlaenaceae. PMID- 10817984 TI - Reconstruction of the Leaves of Two New Species of Pseudosmodingium (Anacardiaceae) from Oligocene Strata of Puebla, Mexico. AB - Leaves of two new plants are reconstructed from their isolated leaflets collected from the Oligocene Los Ahuehuetes locality near Tepexi de Rodriguez in Puebla, Mexico. The leaves of Pseudosmodingium mirandae Ramirez-Garduno et al. are compound imparipinnate with leaflets of variable morphology. The leaflets of five leaf morphotypes vary from narrow elliptic to lanceolate or lorate; they are symmetrical to slightly asymmetrical, with acute to attenuate apex, acute to cuneate base, and entire to serrate margin. Venation is simple pinnate craspedodromous, with secondary veins slightly curved near their base; secondary veins may dichotomize near the margin to become tertiary veins, and intersecondary veins are small and oblique to the secondary veins. A small number of leaflets assigned to Pseudosmodingium terrazasiae Ramirez-Garduno et al. are distinguished from P. mirandae by the leaflet shape, length&rcolon;width ratio, base shape, and apex angle. Morphological comparison of the fossil leaves with leaves of extant species of Anacardiaceae based on numerical analyses indicates a close similarity between P. mirandae and Pseudosmodingium multifolium Rose, while P. terrazasiae is more similar to Pseudosmodingium perniciosum (HBK) Engl. The presence of fossil species with extant relatives that are endemic to Mexico, along with previous reports, indicates that by the Oligocene, some lineages were already in place, although today they form part of the more xeric communities in southern North America. PMID- 10817985 TI - Leaves of Salicaceae (Salix and Populus) from Oligocene Sediments Near Tepexi de Rodriguez, Puebla, Mexico. AB - Eleven new salicaceous taxa based on fossil leaves, seven Salix and four Populus species, are described from the Oligocene Los Ahuehuetes locality, Pie de Vaca Formation, near Tepexi de Rodriguez in Puebla, Mexico. They are characterized by the presence of typical salicoid teeth, camptodromous secondary venation, and elliptic or lanceolate shape. Morphological differences between these new fossil plants and those reported previously from the United States and elsewhere indicate that diversification among these taxa occurred early during their differentiation. Furthermore, the limited morphological similarities of the fossil and extant plants growing naturally in Mexico also indicate a high rate of evolution. Documenting the expansion of high-latitude North American taxa into lower latitudes at the time of deposition of the Pie de Vaca Formation confirms that at least some of the typical boreal taxa were part of dry tropical communities in low-latitude North America and underscores the need to reevaluate the origin of this southern vegetation. PMID- 10817986 TI - Galileo at Io: results from high-resolution imaging. AB - During late 1999/early 2000, the solid state imaging experiment on the Galileo spacecraft returned more than 100 high-resolution (5 to 500 meters per pixel) images of volcanically active Io. We observed an active lava lake, an active curtain of lava, active lava flows, calderas, mountains, plateaus, and plains. Several of the sulfur dioxide-rich plumes are erupting from distal flows, rather than from the source of silicate lava (caldera or fissure, often with red pyroclastic deposits). Most of the active flows in equatorial regions are being emplaced slowly beneath insulated crust, but rapidly emplaced channelized flows are also found at all latitudes. There is no evidence for high-viscosity lava, but some bright flows may consist of sulfur rather than mafic silicates. The mountains, plateaus, and calderas are strongly influenced by tectonics and gravitational collapse. Sapping channels and scarps suggest that many portions of the upper approximately 1 kilometer are rich in volatiles. PMID- 10817987 TI - Io's thermal emission from the Galileo photopolarimeter-radiometer. AB - Galileo's photopolarimeter-radiometer instrument mapped Io's thermal emission during the I24, I25, and I27 flybys with a spatial resolution of 2.2 to 300 kilometers. Mapping of Loki in I24 shows uniform temperatures for most of Loki Patera and high temperatures in the southwest corner, probably resulting from an eruption that began 1 month before the observation. Most of Loki Patera was resurfaced before I27. Pele's caldera floor has a low temperature of 160 kelvin, whereas flows at Pillan and Zamama have temperatures of up to 200 kelvin. Global maps of nighttime temperatures provide a means for estimating global heat flow. PMID- 10817988 TI - A close-up look at Io from Galileo's near-infrared mapping spectrometer. AB - Infrared spectral images of Jupiter's volcanic moon Io, acquired during the October and November 1999 and February 2000 flybys of the Galileo spacecraft, were used to study the thermal structure and sulfur dioxide distribution of active volcanoes. Loki Patera, the solar system's most powerful known volcano, exhibits large expanses of dark, cooling lava on its caldera floor. Prometheus, the site of long-lived plume activity, has two major areas of thermal emission, which support ideas of plume migration. Sulfur dioxide deposits were mapped at local scales and show a more complex relationship to surface colors than previously thought, indicating the presence of other sulfur compounds. PMID- 10817989 TI - Prometheus: Io's wandering plume. AB - Unlike any volcanic behavior ever observed on Earth, the plume from Prometheus on Io has wandered 75 to 95 kilometers west over the last 20 years since it was first discovered by Voyager and more recently observed by Galileo. Despite the source motion, the geometric and optical properties of the plume have remained constant. We propose that this can be explained by vaporization of a sulfur dioxide and/or sulfur "snowfield" over which a lava flow is moving. Eruption of a boundary-layer slurry through a rootless conduit with sonic conditions at the intake of the melted snow can account for the constancy of plume properties. PMID- 10817990 TI - Discovery of gaseous S2 in Io's Pele plume. AB - Spectroscopy of Io's Pele plume against Jupiter by the Hubble Space Telescope in October 1999 revealed absorption due to S2 gas, with a column density of 1.0 +/- 0.2 x 10(16) per square centimeter, and probably also SO(2) gas with a column density of 7 +/- 3 x 10(16) per square centimeter. This SO2/S2 ratio (3 to 12) is expected from equilibration with silicate magmas near the quartz-fayalite magnetite or wustite-magnetite buffers. Condensed S3 and S4, probable coloring agents in Pele's red plume deposits, may form by polymerization of the S2, which is unstable to ultraviolet photolysis. Diffuse red deposits near other Io volcanoes suggest that venting and polymerization of S2 gas is a widespread feature of Io volcanism. PMID- 10817992 TI - Rocks from the mantle transition zone: majorite-bearing xenoliths from malaita, southwest pacific AB - Rocks containing high-pressure mineral assemblages derived from the mantle transition zone between depths of about 400 and 670 kilometers occur as xenoliths and megacrysts on the island of Malaita in the southwest Pacific on the Ontong Java Plateau. Observed ultrahigh pressure mineral chemistries include majorite, calcium- and magnesium-perovskite, aluminous silicate phases, and microdiamond. Based on an empirical barometer, majoritic garnets in these xenoliths record pressures of up to 22 gigapascal. The occurrence of material with perovskite chemistry and several enigmatic aluminous phases indicates pressures of up to 27 gigapascal. Samples were brought to the surface at about 34 million years ago by potassic ultramafic magmas, which evidently originated in the lower mantle. PMID- 10817991 TI - Evidence for the evolution of bdelloid rotifers without sexual reproduction or genetic exchange. AB - The Class Bdelloidea of the Phylum Rotifera is the largest metazoan taxon in which males, hermaphrodites, and meiosis are unknown. We conducted a molecular genetic test of this indication that bdelloid rotifers may have evolved without sexual reproduction or genetic exchange. The test is based on the expectation that after millions of years without these processes, genomes will no longer contain pairs of closely similar haplotypes and instead will contain highly divergent descendants of formerly allelic nucleotide sequences. We find that genomes of individual bdelloid rotifers, representing four different species, appear to lack pairs of closely similar sequences and contain representatives of two ancient lineages that began to diverge before the bdelloid radiation many millions of years ago when sexual reproduction and genetic exchange may have ceased. PMID- 10817993 TI - Molecular computation by DNA hairpin formation. AB - Hairpin formation by single-stranded DNA molecules was exploited in a DNA-based computation in order to explore the feasibility of autonomous molecular computing. An instance of the satisfiability problem, a famous hard combinatorial problem, was solved by using molecular biology techniques. The satisfiability of a given Boolean formula was examined autonomously, on the basis of hairpin formation by the molecules that represent the formula. This computation algorithm can test several clauses in the given formula simultaneously, which could reduce the number of laboratory steps required for computation. PMID- 10817994 TI - Coalescence of single-walled carbon nanotubes AB - The coalescence of single-walled nanotubes is studied in situ under electron irradiation at high temperature in a transmission electron microscope. The merging process is investigated at the atomic level, using tight-binding molecular dynamics and Monte Carlo simulations. Vacancies induce coalescence via a zipper-like mechanism, imposing a continuous reorganization of atoms on individual tube lattices along adjacent tubes. Other topological defects induce the polymerization of tubes. Coalescence seems to be restricted to tubes with the same chirality, explaining the low frequency of occurrence of this event. PMID- 10817995 TI - Suppressing wall turbulence by means of a transverse traveling wave AB - Direct numerical simulations of wall-bounded flow reveal that turbulence production can be suppressed by a transverse traveling wave. Flow visualizations show that the near-wall streaks are eliminated, in contrast to other turbulence control techniques, leading to a large shear stress reduction. The traveling wave can be induced by a spanwise force that is confined within the viscous sublayer; it has its maximum at the wall and decays exponentially away from it. We demonstrate the application of this approach in salt water, using arrays of electromagnetic tiles that produce the required traveling wave excitation at a high efficiency. PMID- 10817996 TI - The spin excitation spectrum in superconducting YBa(2)Cu(3)O(6.85) AB - A comprehensive inelastic neutron scattering study of magnetic excitations in the near optimally doped high-temperature superconductor YBa(2)Cu(3)O(6.85) is presented. The spin correlations in the normal state are commensurate with the crystal lattice, and the intensity is peaked around the wave vector characterizing the antiferromagnetic state of the insulating precursor, YBa(2)Cu(3)O(6). Profound modifications of the spin excitation spectrum appear abruptly below the superconducting transition temperature T(c), where a commensurate resonant mode and a set of weaker incommensurate peaks develop. The data are consistent with models that are based on an underlying two-dimensional Fermi surface, predicting a continuous, downward dispersion relation connecting the resonant mode and the incommensurate excitations. The magnetic incommensurability in the YBa(2)Cu(3)O(6+)(x) system is thus not simply related to that of another high-temperature superconductor, La(2-)(x)Sr(x)CuO(4), where incommensurate peaks persist well above T(c). The temperature-dependent incommensurability is difficult to reconcile with interpretations based on charge stripe formation in YBa(2)Cu(3)O(6+x) near optimum doping. PMID- 10817997 TI - Propagation of seismic ground motion in the Kanto Basin, Japan. AB - The pattern of ground motion for a magnitude 5.7 earthquake near Tokyo was captured by 384 strong ground motion instruments across the Kanto sedimentary basin and its surroundings. The records allow the visualization of the propagation of long-period ground motion in the basin and show the refraction of surface waves at the basin edge. The refracted wave does not travel directly from the earthquake epicenter, but traverses the basin obliquely to the edge. The surface wave inside the basin propagates more slowly than that outside such that the wavefronts separate from each other, and the refracted wave heals the discrepancy in the speed of advance of the wavefronts inside and outside the basin. The refracted arrival is dominant near the edge of the Kanto basin. PMID- 10817998 TI - Evolutionary exploitation of design options by the first animals with hard skeletons. AB - The set of viable design elements available for animals to use in building skeletons has been fully exploited. Analysis of animal skeletons in relation to the multivariate, theoretical "Skeleton Space" has shown that a large proportion of these options are used in each phylum. Here, we show that structural elements deployed in the skeletons of Burgess Shale animals (Middle Cambrian) incorporate 146 of 182 character pairs defined in this morphospace. Within 15 million years of the appearance of crown groups of phyla with substantial hard parts, at least 80 percent of skeletal design elements recognized among living and extinct marine metazoans were exploited. PMID- 10817999 TI - Distinct classes of yeast promoters revealed by differential TAF recruitment. AB - The transcription factor TFIID contains the TATA box binding protein (TBP) and multiple TBP-associated factors (TAFs). Here, the association of TFIID components with promoters that either are dependent on multiple TAFs (TAFdep) or have no apparent TAF requirement (TAFind) is analyzed in yeast. At TAFdep promoters, TAFs are present at levels comparable to that of TBP, whereas at TAFind promoters, TAFs are present at levels that approximate background. After inactivation of several general transcription factors, including TBP, TAFs are still recruited by activators to TAFdep promoters. The results reveal two classes of promoters: at TAFind promoters, TBP is recruited in the apparent absence of TAFs, whereas at TAFdep promoters, TAFs are co-recruited with TBP in a manner consistent with direct activator-TAF interactions. PMID- 10818000 TI - TAF-Containing and TAF-independent forms of transcriptionally active TBP in vivo. AB - Transcriptional activity in yeast strongly correlates with promoter occupancy by general factors such as TATA binding protein (TBP), TFIIA, and TFIIB, but not with occupancy by TBP-associated factors (TAFs). Thus, TBP exists in at least two transcriptionally active forms in vivo. The TAF-containing form corresponds to the TFIID complex, whereas the form lacking TAFs corresponds to TBP itself or to some other TBP complex. Heat shock treatment altered the relative utilization of these TBP forms, with TFIID being favored. Promoter-specific variations in the association of these distinct forms of TBP may explain why only some yeast genes require TFIID for transcriptional activity in vivo. PMID- 10818001 TI - A mutation in PRKAG3 associated with excess glycogen content in pig skeletal muscle. AB - A high proportion of purebred Hampshire pigs carries the dominant RN- mutation, which causes high glycogen content in skeletal muscle. The mutation has beneficial effects on meat content but detrimental effects on processing yield. Here, it is shown that the mutation is a nonconservative substitution (R200Q) in the PRKAG3 gene, which encodes a muscle-specific isoform of the regulatory gamma subunit of adenosine monophosphate-activated protein kinase (AMPK). Loss-of function mutations in the homologous gene in yeast (SNF4) cause defects in glucose metabolism, including glycogen storage. Further analysis of the PRKAG3 signaling pathway may provide insights into muscle physiology as well as the pathogenesis of noninsulin-dependent diabetes mellitus in humans, a metabolic disorder associated with impaired glycogen synthesis. PMID- 10818002 TI - High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. AB - The lungs of cystic fibrosis (CF) patients are chronically infected for years by one or a few lineages of Pseudomonas aeruginosa. These bacterial populations adapt to the highly compartmentalized and anatomically deteriorating lung environment of CF patients, as well as to the challenges of the immune defenses and antibiotic therapy. These selective conditions are precisely those that recent theoretical studies predict for the evolution of mechanisms that augment the rate of variation. Determination of spontaneous mutation rates in 128 P. aeruginosa isolates from 30 CF patients revealed that 36% of the patients were colonized by a hypermutable (mutator) strain that persisted for years in most patients. Mutator strains were not found in 75 non-CF patients acutely infected with P. aeruginosa. This investigation also reveals a link between high mutation rates in vivo and the evolution of antibiotic resistance. PMID- 10818003 TI - Role for rapid dendritic protein synthesis in hippocampal mGluR-dependent long term depression. AB - A hippocampal pyramidal neuron receives more than 10(4) excitatory glutamatergic synapses. Many of these synapses contain the molecular machinery for messenger RNA translation, suggesting that the protein complement (and thus function) of each synapse can be regulated on the basis of activity. Here, local postsynaptic protein synthesis, triggered by synaptic activation of metabotropic glutamate receptors, was found to modify synaptic transmission within minutes. PMID- 10818004 TI - Impaired prion replication in spleens of mice lacking functional follicular dendritic cells. AB - In scrapie-infected mice, prions are found associated with splenic but not circulating B and T lymphocytes and in the stroma, which contains follicular dendritic cells (FDCs). Formation and maintenance of mature FDCs require the presence of B cells expressing membrane-bound lymphotoxin-alpha/beta. Treatment of mice with soluble lymphotoxin-beta receptor results in the disappearance of mature FDCs from the spleen. We show that this treatment abolishes splenic prion accumulation and retards neuroinvasion after intraperitoneal scrapie inoculation. These data provide evidence that FDCs are the principal sites for prion replication in the spleen. PMID- 10818005 TI - Shots in the desert and Gulf war syndrome. Evidence that multiple vaccinations during deployment are to blame is inconclusive. PMID- 10818006 TI - Screening for breast and cervical cancer as a common cause for litigation. A false negative result may be one of an irreducible minimum of errors. PMID- 10818007 TI - A national screening policy for sickle cell disease and thalassaemia major for the United Kingdom. Questions are left after two evidence based reports. PMID- 10818008 TI - Reperfusion in acute myocardial infarction. Ensuring early reperfusion, by whatever means, is the best strategy for now. PMID- 10818009 TI - The GMC: where now? It must put the public first but listen to doctors and be bold with its reforms. General Medical Council. PMID- 10818010 TI - Drug companies cut HIV drug prices in the developing world.